PMID- 8400530 TI - 'The high price of hips'. Whose name is on the bill? AB - Many of hospital's more secret corners have been the subject of television exposes in the past 12 months and it was probably long overdue that the validity of joint replacement should finally come under scrutiny. Public Eye did just this (BBC2, February 26th 1993), investigating the number of revision hip arthroplasties that are becoming necessary in the UK--some 12% at the last report with a projected increase over the next two years to a massive 30%. The likely repercussions from this and the inability of our groaning health service to cope with the influx, are truly staggering and will mean that people waiting for primary replacements will simply wait longer and longer. PMID- 8400531 TI - How recovery helped me to recover. Experiences of hyperhydrosis. PMID- 8400532 TI - The quality of treatment being delivered in today's health service. PMID- 8400533 TI - Revolution in management development. AB - There can be few important areas of working life that have seen more changes in fashion and style than the process of management. Anyone who has been close to management and management education over the last 25 years will have been washed over by a constant stream of new theories, techniques and 'guaranteed solutions' aimed at all aspects of how we manage people, finance and resources. Many of the things coming out of this process have proved valuable and are now permanent features of the way we manage our affairs. PMID- 8400534 TI - Standards for records and record keeping. PMID- 8400535 TI - Mercy ships. A ministry of youth with a mission. PMID- 8400536 TI - Cardiac transplantation (2). Psychological aspects. AB - In my previous article I looked at the quality of life that cardiac transplant patients have both before and after the actual operation. The quality of these patients' lives is, in nearly every case, extremely poor. Most (97%) are unable to work or even do everyday tasks such as housework, cooking and so on. A large number are housebound. Many of them confined to sleeping on the ground floor as climbing stairs is impossible to them. This, along with the knowledge that they face death without a transplant, has a profound effect on their mental state. PMID- 8400537 TI - Is there a place for nursing models in theatre nursing? AB - Nursing models are described as theoretical frameworks, or concepts, which represent what nursing is. A concept is 'something conceived in the mind, an idea, covering many similar things derived from study of particular instances'. You might ask, why do we need a model to explain what we do? But nursing has been notoriously difficult to define. Two definitions often cited are: 'The unique function of the nurse is to assist the individual, sick or well, in the performance of those activities contributing to health or its recovery (or a peaceful death) that he would perform unaided, if he had the necessary strength, will or knowledge. And to do it in such a way as to help him gain independence as rapidly as possible'. and: 'Nursing consists essentially of acts of helping or assistance ... the conditions which validate nursing action are lack of strength or will or knowledge. The nursing role, therefore, takes on physical and psychological assistance and its teaching function'. These provide us with a basis to initiate our ideas of nursing, but provide few answers to the 'how' of nursing. PMID- 8400538 TI - Beliefs, values, attitudes and nursing models. AB - 'I only came into theatres to get out of the way of the patients.' This remark was heard in the staff dining room on a busy weekday lunchtime. It is in the light of such conversations that I consider the issue of theatre nursing and nursing models, particularly in relation to examining beliefs and values of the sub-culture. According to Hilgard and Atkinson values are beliefs which an individual views as positive attributes. Many nurses reading this article would not value the above statement. However, when the definition of 'belief' is analysed it is suggested as being 'a statement that a person thinks is true'. Such a statement as this portrays a negative attitude towards patients requiring nursing intervention. PMID- 8400539 TI - 'Awake for an operation'. AB - It seems likely that every single patient coming into hospital for surgery feels some apprehension, if not fear. Part of this concern comes from what is unknown, some from the prospect of pain and perhaps from the prospect of 'something going wrong'. PMID- 8400540 TI - Using Orem's Model. AB - My actual knowledge of care plans and nursing models was non-existent until my conversion course to RGN in 1989. I found it difficult to grasp and at times a paper exercise. A care plan should be used as a link between the outpatient's nurse and ward nurses involved with the patient's primary care and the personnel in theatre responsible for the patient. The purpose of nursing documentation is to show that care has been planned for each individual patient; to inform nursing staff and others involved in delivering care; to record changes in the patient's condition or care; to maintain continuity of care. PMID- 8400541 TI - Reflections--where does it hurt? PMID- 8400542 TI - Nitrous oxide: your health not theirs. AB - At long last the Health and Safety Executive are introducing an occupational exposure standard for nitrous oxide, enflurane, halothane and isoflurane. Fifteen years ago (1977) the National Institute of Occupational Safety and Health in the United States set a threshold limit for nitrous oxide at 25 parts per million (ppm) for an eight hour exposure. On the 1st January 1994 the British standard for nitrous oxide through the Health and Safety Executive will be 100 ppm for an eight hour exposure, four times higher than the American standard. PMID- 8400543 TI - Postoperative nausea and vomiting. AB - Postoperative nausea and vomiting (PONV) is a widespread and often underestimated problem that invariably falls onto the shoulders of nursing staff. In order to find out how nurses view this condition, a major survey was conducted amongst members of the National Association of Theatre Nurses. PMID- 8400544 TI - Carbohydrate signals in metastasis and prognosis of human carcinomas. PMID- 8400545 TI - The family of metazoan metal-independent beta-galactoside-binding lectins: structure, function and molecular evolution. AB - Animal metal-independent beta-galactoside-binding lectins were initially found in vertebrates, but they have recently been isolated from much lower invertebrates, such as nematode and sponge, as well. Further, an eosinophilic lysophospholipase associated with various inflammatory reactions was very recently found to be a new member of this protein family. It appears that beta-galactoside-binding lectins and some non-lectin proteins form a superfamily whose members are widely distributed from vertebrates to invertebrates. From the viewpoints of protein architecture, the superfamily members can be subdivided into three types; i.e. 'proto type' (the relatively well-studied 14 kDa lectins), 'chimera type' (29-35 kDa lectins also known as epsilon BP/CBP35/Mac2/laminin-binding protein) and 'tandem-repeat type' (a newly found nematode 32 kDa lectin). Comparison of their amino acid sequences and mutagenesis studies have suggested the functional importance of some conservative hydrophilic residues (His44, Asn46, Arg48, Glu71 and Arg73 of human 14 kDa lectin). Several non-charged residues (Gly14, Phe45, Pro47, Phe49, Val59, Trp68, Pro78 and Phe79) are also well conserved, and are probably important to maintain the structural framework of these proteins. A consideration of molecular evolution suggests that lectins belonging to this family probably existed in the Precambrian era. Ubiquitous occurrence of these homologous lectins with shared sugar specificity suggests that they are involved in 'essential minimum' functions of multicellular animals, possibly in cooperation with their partner glycoconjugates. PMID- 8400546 TI - N-acetylglucosamine-binding proteins on Plasmodium falciparum merozoite surface. AB - Plasmodium falciparum merozoite surface is specifically labelled with a neoglycoprotein bearing N-acetylglucosamine (GlcNAc) residues in a sugar dependent manner, as shown by affinity cytochemistry in fluorescence and electron microscopy. To ascertain the nature of the sugar receptor, merozoite proteins were blotted and tested by a two-step method using biotinylated GlcNAc-bovine serum albumin (BSA) and streptavidin-peroxidase conjugate. Three parasite proteins were specifically revealed and designated as Pf 120, Pf 83 and Pf 45 GlcNAc-binding proteins. These proteins bind to a gel substituted with GlcNAc and are specifically eluted with 300 mM GlcNAc. Using a rabbit antiserum raised against Pf 83, the Pf 120 GlcNAc-binding protein, in addition to Pf 83, was labelled by Western blotting. Comparative analyses with an antibody against the Pf 83 MSP derived from the P. falciparum merozoite surface protein (Pf MSP) indicated that the Pf 83 GlcNAc-binding protein is not related to the fragment of the Pf MSP antigen. Similarly, the Pf 83 GlcNAc-binding protein is not related to the apical membrane antigen 1 (AMA 1) which also has the same molecular mass. Therefore the Pf 120, Pf 83 and Pf 45 GlcNAc-binding proteins which are located on the merozoite surface and recognize GlcNAc residues could be involved in the binding of merozoites to the glycoconjugates of the surface of the red blood cells. PMID- 8400547 TI - Light and heavy lysosomes: characterization of N-acetyl-beta-D-hexosaminidase isolated from normal and I-cell disease lymphoblasts. AB - We previously reported that I-cell disease lymphoblasts maintain normal or near normal intracellular levels of lysosomal enzymes, even though N-acetylglucosamine 1-phosphotransferase activity is severely depressed or absent (Little et al., Biochem. J., 248, 151-159, 1987). The present study, employing subcellular fractionation on colloidal silica gradients, indicates that both light and heavy lysosomes isolated from I-cell disease and pseudo-Hurler polydystrophy lymphoblasts possess normal specific activity levels of N-acetyl-beta-D hexosaminidase, alpha-D-mannosidase and beta-D-glucuronidase. These current findings are in contrast to those of cultured fibroblasts from the same patients, where decreased intralysosomal enzyme activities are found. Column chromatography on Ricinus communis revealed that N-acetyl-beta-D-hexosaminidase in both heavy and light I-cell disease lysosomal fractions from lymphoblasts possesses an increased number of accessible galactose residues (30-50%) as compared to the enzyme from the corresponding normal controls. Endo-beta-N-acetylglucosaminidase H treatment of N-acetyl-beta-D-hexosaminidase from the I-cell lysosomal fractions suggests that the majority of newly synthesized high-mannose-type oligosaccharide chains are modified to complex-type carbohydrates prior to being transported to lysosomes. This result from lymphoblasts differs from previous findings with fibroblasts, where N-acetyl-beta-D-hexosaminidase from I-cell disease and pseudo Hurler polydystrophy lysosomes exhibited properties associated with predominantly high-mannose-type oligosaccharide chains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400548 TI - Glycosphingolipid inhibition of the adhesion of thrombin-activated platelets to surfaces is potentiated by albumin. AB - Previous studies have shown that exogenous glycosphingolipids (GSLs) inhibit the adhesion of thrombin-activated platelets (TAP) to polystyrene plates coated with various RGD-ligands (where RGD is the peptide sequence Arg-Gly-Asp), suggesting that GSLs can modulate the platelet integrin receptor glycoprotein IIb-IIIa. However, albumin was always used as a plastic surface-blocking agent in these studies. In order to evaluate the role of albumin in these experiments, we studied the effect of various GSLs and albumin on the interaction between TAP and hydrophobic surfaces in a solid-phase assay using indium-111-labelled platelets and polystyrene plates. TAP (10(8) platelets/ml) adhered to polystyrene (half saturation time 40 +/- 3 min) with a maximal adhesion density of 56 +/- 1 x 10(3) platelets/mm2. Platelet adhesion was only slightly affected (< 11% inhibition) by immobilized bovine serum albumin, immobilized mixed bovine brain gangliosides (MBG) or fluid-phase MBG. In contrast, fluid-phase MBG was an effective inhibitor of platelet adhesion to polystyrene (> 46% inhibition), but only after albumin was first immobilized to the plate. Covering albumin-coated polystyrene with MBG, followed by washing, was as effective as fluid-phase MBG at inhibiting platelet adhesion, thus indicating that a ganglioside-albumin interaction at the polystyrene surface was responsible for effective inhibition. When purified GSLs were substituted for MBG, it was found that all those tested (GT1b, GD1a, GM1, asialo GM1 and globoside) had similar inhibitory activity. PMID- 8400549 TI - Investigation of sucrose synthase from rice for the synthesis of various nucleotide sugars and saccharides. AB - The unique character of the plant glucosyltransferase sucrose synthase, to catalyse in vitro the synthesis and cleavage of sucrose under appropriate conditions, can be exploited for the enzymatic synthesis of carbohydrates. The present paper describes the potential utilization of sucrose synthase from rice for the enzymatic synthesis of activated sugars and saccharides. In the cleavage reaction of sucrose, the nucleoside diphosphates can be used in the order UDP > TDP > ADP > CDP > GDP to obtain the corresponding activated glucoses. In batch reactions, > 90% conversion of UDP and TDP could be achieved. Substituting different di- and trisaccharides for sucrose in the cleavage reaction with UDP 2 deoxysucrose was the most promising substrate. Sucrose synthase was combined with UDP-galactose 4'-epimerase and beta 1-4 galactosyltransferase to synthesize N acetyllactosamine with in situ regeneration of UDP-glucose. In the synthesis reaction of sucrose synthase, different donor (UDP-sugars) and acceptor substrates were investigated. UDP-N-acetylglucosamine and UDP-xylose could be used in combination with fructose as acceptor. D-Xylulose, D-tagatose, D-lyxose, D-psicose, L-sorbose, D-mannose, L-arabinose, 1,6 anhydroglucose, lactulose, raffinose and isomaltulose can serve as acceptors for UDP-glucose. PMID- 8400550 TI - Biosynthesis of asparagine-linked oligosaccharides in Saccharomyces cerevisiae: the alg2 mutation. AB - In the yeast Saccharomyces cerevisiae, the alg2 mutation causes temperature sensitive growth and abnormal accumulation of the lipid-linked oligosaccharide Man2GlcNAc2-PP-Dol (Jackson et al., Arch. Biochem. Biophys., 272, 203-209, 1989; Huffaker and Robbins, Proc. Natl. Acad. Sci. USA, 80, 7466-7470, 1983). A gene having the function and genomic location of ALG2 was cloned from libraries based on the multicopy plasmid YEp24 and on the centromere plasmid YCp50. Alg2 mutants transformed with plasmids containing ALG2 regained the capacity to grow and to synthesize lipid-linked oligosaccharides normally at the previously non permissive temperature. ALG2 was essential for viability in haploid and diploid yeast. The ALG2 gene was transcribed into a single mRNA of 1.7 kb in size. The stability of ALG2 mRNA, assessed after thermal inactivation of RNA polymerase II in an rpb1-1 mutant (Herrick et al., Mol. Cell. Biol., 10, 2269-2284, 1990) was very low, with a t1/2 of < 5 min. The ALG2 transcript accumulation was growth dependent, and it was at least an order of magnitude lower in stationary phase cells compared to exponentially growing cells. The putative translation product of ALG2 contained a potential dolichol recognition domain similar to that found in all three glycosyltransferases of the lipid-linked pathway that have been sequenced.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400551 TI - Identification and oligosaccharide structure analysis of rhodopsin glycoforms containing galactose and sialic acid. AB - The N-linked oligosaccharides of frog (Rana pipiens) rhodopsin were analysed by sequential exoglycosidase digestion and gel filtration chromatography, following reductive tritiation. In addition, selected tryptic glycopeptides obtained from frog retinal rod outer segment membranes were examined by electrospray mass spectrometry (ES-MS), fast atom bombardment mass spectrometry (FAB-MS), amino acid sequence and composition analysis, and carbohydrate composition analysis. The amino acid sequence data demonstrated that the glycopeptides were derived from rhodopsin and confirmed the presence of two N-glycosylation sites, at residues Asn2 and Asn15. The predominant glycan (approximately 60% of total) had the structure GlcNAc beta 1-2Man alpha 1-3(Man alpha 1-6) Man beta 1-4GlcNAc beta 1-4GlcNAc-(Asn), with the remaining structures containing 1-3 additional hexose residues, as reported previously for bovine rhodopsin. Unlike bovine rhodopsin, however, a sizable fraction of the total glycans of frog rhodopsin also contained sialic acid (NeuAc), with the sialylated oligosaccharides being present exclusively at the Asn2 site. FAB-MS analysis of oligosaccharides released from the Asn2 site gave, among other signals, an abundant quasimolecular ion corresponding to a glycan of composition NeuAc1Hex6HexNAc3 (where Hex is hexose and HexNAc is N-acetylhexosamine), consistent with a hybrid structure. The potential biological implications of these results are discussed in the context of rod outer segment membrane renewal. PMID- 8400552 TI - Glycan localization within the human interphotoreceptor matrix and photoreceptor inner and outer segments. AB - Glycoconjugates are likely to be of fundamental importance in the complex interactions between photoreceptors and the retinal pigment epithelium, but few have been characterized, especially in human tissue. As a preliminary step towards determining their biological functions in health and disease, a lectin based histochemical study of the glycan expression of human outer retina was performed on glutaraldehyde-fixed, semi-thin, resin-embedded sections. The interphotoreceptor matrix and photoreceptor plasma-lemmata expressed complex bisected and non-bisected biantennary and/or triantennary N-glycans. In addition, both the rod and cone outer segments bound strongly Galanthus nivalis agglutinin (which binds terminal Man alpha 1, 3Man-) and the rod outer segments bound selectively the isolectin II of Bandeiraea simplicifolia (which binds terminal GlcNAc-). The cilia of the rods and cones were labelled selectively with Glycine max agglutinin after sialidase pretreatment. Four putative glycan outer sequences were identified within the interphotoreceptor matrix: (i) sialylated glycans with subterminal GalNAc alpha 1,3GalNAc-sequences; (ii) a sialylated type with subterminal N-acetyl-lactosamine residues; (iii) Gal beta 1,3GalNAc alpha 1- residues which were substituted with sialic acid except in the cone matrix sheath; (iv) GalNAc alpha 1,6Gal beta 1- residues which were substituted in part with sialic acid. The sialic acid expression throughout was predominantly of the 2,3-linked form with lesser amounts of 2,6-linkage, and rod-associated structures (including the surrounding interphotoreceptor matrix) were labelled more strongly with the sialic acid-binding lectins than cone-associated structures (including the cone matrix sheath). PMID- 8400553 TI - Relative characteristics of MR angiography and competing vascular imaging modalities. AB - This article reviews the general characteristics of several vascular imaging modalities with the purpose of identifying the distinguishing features of magnetic resonance (MR) angiography. Brief discussions of conventional x-ray film angiography, intravenous and intraarterial digital subtraction angiography (DSA), duplex and color Doppler flow ultrasound (US), computed tomographic (CT) angiography, transesophageal and intravascular US, angioscopy, and MR angiography are presented. The advantages and disadvantages of each are discussed. The general attributes and image quality features of MR angiography, intraarterial DSA, CT angiography, and US are compared. It is concluded that no single imaging modality will presently suffice for all purposes. Because of its noninvasiveness, rapidly improving image quality, and ability to directly provide velocity information, MR angiography is likely to play a role in an increasing number of clinical applications. PMID- 8400554 TI - Flow velocity quantification in human coronary arteries with fast, breath-hold MR angiography. AB - Measurement of coronary artery flow velocities has, until now, largely required the use of invasive technologies. The authors have implemented a breath-hold magnetic resonance (MR) angiography technique for depicting the coronary arteries and for quantifying flow velocities. The method was tested in flow phantoms and then applied to a series of subjects: 11 subjects were studied at rest, and four were studied before and during pharmacologic stress induced by intravenous adenosine. Flow velocities at rest in the midportion of the right coronary artery were 9.9 cm/sec +/- 3.5 (n = 12); in the proximal left anterior descending coronary artery, they were significantly higher, measuring 20.5 cm/sec +/- 5.2 (n = 6). With adenosine, flow velocities typically increased at least fourfold. The authors conclude that noninvasive measurement of coronary artery flow velocities is feasible with MR angiography; this method may prove useful for determining the physiologic significance of coronary artery stenosis. PMID- 8400555 TI - Comparison of cerebral artery blood flow measurements with gated cine and ungated phase-contrast techniques. AB - Cine phase-contrast (PC) magnetic resonance (MR) pulse sequences have been used to measure blood flow in a variety of vessels. Because the cine PC sequence is time-consuming, this prospective study was undertaken to compare it with an ungated PC technique for measuring average blood flow in individual cerebral arteries to potentially achieve substantial time savings. The following cerebral arteries were studied in 10 healthy volunteers: carotid, basilar, middle cerebral, anterior cerebral, and posterior cerebral. Imaging planes were placed perpendicular to the vessel of interest, and velocity encoding, ranging from 40 to 250 cm/sec, was matched to individual arteries. Good correlation between cine and ungated PC blood flow measurements was obtained for both high- and low-flow vessels, with an overall correlation coefficient of .978. The ungated PC sequence, because of its short imaging time, allows measurement of the blood volume flow rate in the circle of Willis in approximately 20 minutes, a clinically acceptable time. PMID- 8400556 TI - MR imaging of silicone gel-filled breast implants in vivo with a method that visualizes silicone selectively. AB - The lack of an adequately sensitive method for detecting silicone leakage and reported serious complications due to silicone leakage were cited as justification by the U.S. Food and Drug Administration for imposing the current restrictions on silicone gel-filled breast prostheses. The authors report a new magnetic resonance imaging method for visualizing silicone leakage: the silicone only sequence (SOS). The method uses the conventional STIR (short-inversion-time inversion-recovery) technique combined with a 1331 radio-frequency pulse train widely used for water suppression in spectroscopy. With the SOS, silicone can be imaged while signals from fat and water are suppressed. The authors used the SOS to image phantoms and normal and ruptured silicone gel-filled breast prostheses. PMID- 8400557 TI - Localized proton MR spectroscopy of the brain in children. AB - Small-voxel (3.0-8.0 cm3), magnetic resonance (MR) imaging-guided proton MR spectroscopy was performed in 54 patients (aged 6 days to 19 years) with intracranial masses (n = 16), neurodegenerative disorders (n = 34), and other neurologic diseases (n = 4) and in 23 age-matched control subjects without brain disease. A combined short TE (18 msec) stimulated-echo acquisition mode (STEAM) and long TE (135 and/or 270 msec) spin-echo point-resolved spatially localized spectroscopy (PRESS) protocol, using designed radio-frequency pulses, was performed at 1.5 T. STEAM spectra revealed short T2 and/or strongly coupled metabolites; prominent resonances were obtained from N-acetyl aspartate (NAA), choline-containing compounds (Cho), and total creatine (tCr). Lactate was well resolved with the long TE PRESS sequence. Intracranial tumors were readily differentiated from cerebrospinal fluid (CSF) collections. All tumors showed low NAA, high Cho, and reduced tCr levels. Neurodegenerative disorders showed low or absent NAA levels and enhanced mobile lipid, glutamate and glutamine, and inositol levels, consistent with neuronal loss, gliosis, demyelination, and amino acid neurotoxicity. Preliminary experience indicates that proton MR spectroscopy can contribute in the evaluation of central nervous system abnormalities of infants and children. PMID- 8400558 TI - Mn-DPDP-enhanced MR imaging of malignant liver lesions: efficacy and safety in 20 patients. AB - Twenty patients with malignant liver lesions underwent magnetic resonance (MR) imaging with manganese (II) DPDP [N,N'-dipyridoxylethylenediamine-N,N'-diacetate 5,5'-bis(phosphate)] to evaluate the safety and efficacy of the contrast agent. In two groups of 10 patients each, 5 mumol/kg Mn-DPDP was administered intravenously (3 mL/min) at a concentration of either 50 or 10 mumol/mL. T1- and T2-weighted images were obtained with a 1.5-T imager. Six patients reported a total of eight instances of side effects (flush, feeling of warmth, metallic taste) of which seven occurred at the 50 mumol/mL concentration. A significant decrease in alkaline phosphatase levels 2 hours after injection was recorded. On T1-weighted images, the 10 mumol/mL formulation yielded significantly greater increases in contrast-to-noise ratio (79.8%-137.5%) than the 50 mumol/mL formulation (46.2%-86.6%). In a blinded reader study of 10 patients with one to five lesions each, no lesion was missed on Mn-DPDP--enhanced T1-weighted images; however, four false-positive foci were identified. The authors conclude that slow administration of 5 mumol/kg Mn-DPDP at a concentration of 10 mumol/mL is safe and efficient enough to proceed to further clinical trials. PMID- 8400559 TI - Automated myocardial edge detection on MR images: accuracy in consecutive subjects. AB - The authors previously demonstrated the feasibility of graph-searching-based automated edge detection in cardiac magnetic resonance (MR) imaging. To further assess the clinical utility of this method, unselected images from 11 consecutive subjects undergoing clinically indicated (except for one healthy volunteer) short axis spin-echo MR imaging were analyzed. A total of 142 images from the 11 subjects, encompassing the left ventricle from apex to outflow tract, were analyzed. The computer algorithm correctly identified complete endocardial and epicardial contours in 121 of 142 images (85%). Correlations between observer traced and computer-derived epicardial areas for all images were good (r = .71 for epicardium, r = .83 for endocardium); they improved for a subset of higher quality images (r = .82 for epicardium, r = .92 for endocardium). The authors conclude that the current data further support the usefulness of computer digital image processing in geometric analysis of cardiac MR image data. PMID- 8400560 TI - Adrenal tissue characterization with 0.5-T MR imaging: value of T2*-weighted images. AB - The authors investigated the value of magnetic resonance (MR) imaging at 0.5 T for distinguishing adrenal adenomas from adrenal metastases. The series included 23 adrenal adenomas (18 nonhyperfunctioning, five hyperfunctioning) and 23 adrenal metastases from various organs. Adrenal tumor-liver signal intensity ratios on T1-, T2-, and T2*-weighted images were calculated for adrenal tissue characterization. Adrenal adenomas were more precisely distinguished from adrenal metastases on T2*-weighted images (21 of 23, 91%) than on T2-weighted images (15 of 23, 65%). T1-weighted images were not useful for this distinction. In conclusion, T2*-weighted images were better than routine T2-weighted images for distinguishing adrenal adenomas from adrenal metastases. It can be postulated that the total signal intensity of adrenal adenomas, which contain some fat components, decreased on T2*-weighted images because of an out-of-phase effect. PMID- 8400561 TI - Perfusion and diffusion MR imaging of thromboembolic stroke. AB - A carotid embolic stroke model in rats was studied with a combination of diffusion- and perfusion-sensitive magnetic resonance (MR) imaging at 4.7 T. Capillary blood deoxygenation changes were monitored during formation of focal ischemia by acquiring multisection magnetic susceptibility-weighted echo-planar images. A signal intensity decrease of 7% +/- 3 in ischemic brain (1% +/- 2 in normal brain) was attributable to a T2* decrease due to increased blood deoxygenation, which correlated well with subsequently measured decreases in the apparent diffusion coefficient. The same multisection methods were used to track the first-pass transit of a bolus of dysprosium-DTPA-BMA [diethylenetriaminepentaacetic acid-bis(methylamide)] to assess relative tissue perfusion before and after stroke and after treatment with a thrombolytic agent. Analysis of contrast agent transit profiles suggested a total perfusion deficit in ischemic tissue and essentially unchanged perfusion in normal brain tissue after stroke. PMID- 8400562 TI - First pass of an MR susceptibility contrast agent through normal and ischemic heart: gradient-recalled echo-planar imaging. AB - Gradient-recalled echo-planar magnetic resonance (MR) imaging was used to monitor the first pass of a magnetic susceptibility contrast agent through the heart of normal rats and rats subjected to 60-minute occlusion of the anterior branch of the left main coronary artery. Each animal (six normal and six ischemic) received four doses (0.05, 0.1, 0.15, and 0.2 mmol/kg) of Dy-DTPA-BMA [diethylenetriaminepentaacetic acid-bis(methylamide)] administered as a bolus volume of 1.0 mL/kg. In both normal and ischemic rats, signal intensity in nonischemic myocardium was reduced by the contrast agent in a dose-dependent manner. Signal intensity in the ischemic zone was reduced much less, so that at a contrast agent dose of 0.1 mmol/kg or greater the ischemic zone was clearly defined as a high-intensity zone on echo-planar images. Plots of the change in the apparent T2* relaxation rate (delta R2*) during the peak bolus effect versus injected dose were well fit by straight lines for normal, nonischemic, and ischemic myocardium but not for blood in the left ventricle. No difference was seen between myocardial response in normal animals and in nonischemic regions in animals with coronary artery occlusion. These findings suggest that the contrast agent-induced changes in tissue T2* are monoexponential and support the idea that data derived from contrast transit studies may be useful for calculation of myocardial blood flow. PMID- 8400563 TI - Theoretical analysis of gadopentetate dimeglumine enhancement in T1-weighted imaging of the brain: comparison of two-dimensional spin-echo and three dimensional gradient-echo sequences. AB - By using a theoretical model, the signal difference-to-noise ratios between simulated lesions and normal white matter and gray matter were calculated as a function of lesion concentration of gadopentetate dimeglumine (GD) for two dimensional (2D) T1-weighted spin-echo (SE), three-dimensional (3D) steady-state spoiled gradient-echo (GRE) (FLASH [fast low-angle shot]), and 3D magnetization prepared rapid gradient echo (MP-RAGE) pulse sequences. The 3D GRE sequences provided greater contrast enhancement at relatively high [GD], and the 2D SE sequence demonstrated greater enhancement and a higher rate of enhancement at low [GD]. The results predict that at low [GD], certain lesions could probably be detected with the 2D SE sequence but possibly not with one or both of the 3D GRE sequences. At high [GD], certain lesions could probably be detected with one or both of the 3D GRE sequences but possibly not with the 2D SE sequence. This provides a potential explanation for the clinical observation that certain contrast agent-enhanced lesions appear less conspicuous on 3D GRE images than on 2D SE images and vice versa. Modified parameter values were derived for the 3D FLASH and 3D MP-RAGE sequences that are predicted to produce contrast enhancement behavior equivalent or superior to that of a conventional 2D SE sequence. PMID- 8400564 TI - MR monitoring during cryotherapy in the liver: predictability of histologic outcome. AB - For well-controlled application of cryotherapy to focal liver lesions, real-time monitoring is necessary to limit the final necrotic effect in the treated area. In this study, near real-time magnetic resonance (MR) monitoring images of normal rabbit liver were obtained during the freezing procedure. The MR-monitored, freezing-induced lesions were followed with MR images for up to 3 weeks. Corresponding histologic samples were also obtained over the same time period. Our results indicate that MR images obtained during the freezing procedure can adequately depict the area of final necrosis. Furthermore, histologic changes at each stage of lesion development correlated well with MR signal intensities on follow-up images. With the development of an MR-compatible cryogen probe, MR imaging may prove to be a robust method for monitoring, controlling, and following up cryotherapy in the liver. PMID- 8400565 TI - Ghost phase cancellation with phase-encoding gradient modulation. AB - Motion artifacts are a dominant cause of magnetic resonance image quality degradation. Periodic or nearly periodic motion results in image replicates of the moving structures in spin-warp Fourier imaging. The replicates, or ghosts, propagate in the image in the phase-encoding, or y, direction. These ghosted images can be considered to consist of the time-averaged spin density I0 and a ghost mask g. A set of j ghosted images Ij may be acquired in which the ghost mask is intentionally phase shifted by varying amounts relative to I0 with interleaved acquisitions that have shifted phase-encoding orders or by acquiring multiple images during a single readout period in the presence of an oscillating phase-encoding gradient. The resulting complex images Ij have the same time averaged spin density I0 but have ghost contributions gj that, on a pixel-by pixel basis, trace part of a circle around I0. The source images Ij can then be used to estimate I0. Simulations and experiments with the phase-encoding gradient modulation method show good general ghost suppression for a variety of quasi periodic motion sources including both respiratory-type artifacts and flow artifacts. The primary limitation of the method is the need for rapid gradient switching. PMID- 8400566 TI - Evaluation of hepatocyte-specific paramagnetic contrast media for MR imaging of hepatitis. AB - The hepatocyte-specific paramagnetic magnetic resonance (MR) contrast agents manganese-DPDP [N,N'-dipyridoxylethylenediamine-N,N'-diacetate 5,5'bis (phosphate)] and gadobenate dimeglumine were used for diagnosing chemically induced hepatitis in rats. Ex vivo liver tissue relaxation times and in vivo MR image signal-to-noise ratios were compared before and after contrast agent administration. Ex vivo relaxometry and in vivo MR imaging showed that Mn-DPDP enhanced normal and diseased livers to the same degree at all time points from 5 to 120 minutes. Gadobenate dimeglumine showed reduced T1 and T2 enhancements in hepatitis relative to those of normal liver, in the early phase (5-30 minutes). However, these effects are offsetting, and as a result, MR imaging failed to allow distinction of diseased from normal livers. This surprising result observed in vivo was in fact predicted by applying the Bloch equation to our ex vivo data. Our results show that detection and quantitation of hepatitis with MR imaging enhanced with paramagnetic cell-specific contrast agents will be more difficult than anticipated. PMID- 8400567 TI - Susceptibility effects in porous media in the presence of flow. AB - Magnetic resonance (MR) imaging of microvascular flow is an important topic in biomedicine because it permits access to the functional state of a biologic system. The internal heterogeneous magnetic field due to susceptibility differences within tissues is one of the factors that can affect signal intensity. A glass bead phantom simulating a porous medium was used to experimentally study the effect of the internal magnetic field on MR flow measurements. A physical model was developed to simulate the paths of the moving spins and the local magnetic field distribution in the medium to estimate the signal intensity with spin-phase analysis. The susceptibility variation inside the glass bead phantom was estimated by comparing the simulation results with the experimental data. Experiments were also performed with a tissue-simulating phantom, and the results indicated that the effect of the internal field inhomogeneity on in vivo microvascular flow measurements was negligible. PMID- 8400568 TI - Differentiation between hemangiomas and cysts of the liver with nonenhanced MR imaging: efficacy of T2 values at 1.5 T. AB - The authors studied the utility of non-contrast-agent-enhanced magnetic resonance (MR) imaging for differentiating cysts and cavernous hemangiomas of the liver. Nineteen patients with hemangiomas (51 lesions) and 16 with cysts (30 lesions) were studied with a 1.5-T MR imager. T2 values were calculated with the two-point method to evaluate the efficacy of T2 values in the differentiation between hemangiomas and cysts of the liver. For lesions larger than 1 cm, the mean T2 value of cysts (306 msec +/- 156) was significantly longer than that of hemangiomas (113 msec +/- 15) (P < .0001); there was no overlap of the ranges for T2 values of hemangiomas and cysts. All cysts larger than 1 cm could be differentiated from hemangiomas by using a threshold T2 value of 140 msec. This study suggests that calculated T2 values permit differentiation between hemangiomas and cysts larger than 1 cm at 1.5 T. PMID- 8400569 TI - Reduction of ringing and blurring artifacts in fast spin-echo imaging. AB - A simple method was devised to reduce ringing and blurring artifacts caused by discontinuous T2 weighting of k-space data in fast spin-echo magnetic resonance (MR) imaging. The method demodulates the weighting function along the phase encoding direction by using multiple T2 values derived from a set of non-phase encoded echoes obtained from an extra excitation. The performance of this method was evaluated by computer simulations and experiments, which confirmed its capability of effectively reducing or, in some cases, even completely removing the ringing and blurring artifacts. The results also show that the proposed method produces better results than other artifact reduction methods. The method is particularly useful at high magnetic field strengths (7.1-9.4 T) and with strong gradients (> 20 G/cm) used in MR microscopy, in which the apparent T2 values are short for most tissues. The authors expect that the proposed method will find useful applications in various fast spin-echo pulse sequences. PMID- 8400570 TI - An epidemic model with fatal risk. AB - A new epidemic model is developed that describes the spread of an infectious disease with fatal risk. The model includes as extreme cases the standard general epidemic process (for diseases that confer immunity after infection) and the fatal epidemic process (for diseases that lead inevitably to death). Both deterministic and stochastic versions are examined. First, attention is paid to the temporal and final states of the epidemic. Then the existence of a threshold phenomenon and the qualitative effect of the fatal risk are investigated. PMID- 8400571 TI - The "deterministic simple epidemic" unmasked. AB - We derive a simple but surprising connection between the "deterministic epidemic with recovery" and the spatial birth-and-death process, and discuss its implications for the use of nonlinear differential equation models for the spatial spread of epidemics. This paper consists of a manuscript dating back to 1977, which has been widely referred to in the literature but not previously published, and its more recent extension to include recovery/death in the processes considered, with some additional comments. To aid reference, we retain the original title. PMID- 8400572 TI - Deterministic and stochastic models for the seasonal variability of measles transmission. AB - Fine and Clarkson used a discrete-time epidemic model with variable transmission parameter to analyze measles data for England and Wales for 1950-1965, during the time of biennial epidemics. Their model seems to provide a convincing fit when its parameters are estimated from these data. In particular, they obtained nearly equal estimates for the variable transmission parameter from the widely different data for epidemic and nonepidemic years. They did not, however, study the dynamics of their model. In this paper we make such a study, and find that the model is unsatisfactory in some important respects. It has a unique equilibrium (to be exact, a solution showing only seasonal variation), and neutrally stable oscillations around this equilibrium are possible, but with period approximately 3 years, not 2. (The model also has an exact 3-year cycle, while for intermediate initial values it shows unstable "chaotic" behavior.) If we vary the amount of variation in the transmission parameter, from no variation to 2.5 times the observed amount, the main periodicity remains approximately 3 years, though the complexity of behavior changes, increasing as the amount of variation increases. A continuous-time version of the model also has an equilibrium and a 3-year cycle, the main difference being that these are stable rather than neutrally stable. A stochastic version shows that simulations starting close to either the equilibrium or the 3-year cycle have a high probability of remaining close to the corresponding deterministic solutions for at least 20-30 years. The oscillatory features of the model are thus reasonably robust against stochastic fluctuations. We conclude that the simple homogeneous mixing model cannot be adapted to provide an adequate fit to the data, especially in explaining the observed biennial cycles. It seems likely that the pattern of seasonal variation found by Fine and Clarkson is essentially correct, but for accurate modeling of measles we need also to take into account heterogeneities of mixing in the population, especially those due to age and space. PMID- 8400573 TI - Eradication of helminthic infections. AB - The eradication of helminthic infections described by models of the Nasell-Hirsch type is investigated. Simple criteria are proposed for comparing the efficiency of different modes of eradication. PMID- 8400574 TI - A review and synthesis of the HIV/AIDS epidemic as a multi-stage process. AB - As our understanding of the dynamics of the transmission of the HIV virus toward diagnosis of AIDS increases, numerous models are being developed to help explain the underlying mechanism. We show that many of the models established so far in the literature are examples of a general class of multi-stage epidemic models that are themselves right-shift processes as developed in 1969 by Severo. We focus attention on those models that can be described as three-stage, four-stage, five-stage, etc. models. PMID- 8400575 TI - Large deviations for discrete-time epidemic models. AB - A sufficient condition is presented under which a family of discrete-time epidemic models follows a large deviation principle. Following Freidlin and Wentzell (1984), we study exit time from a domain for such a family of discrete time epidemic models. PMID- 8400576 TI - On the duration of a Maki-Thompson epidemic. AB - In a Maki-Thompson model for the spread of a rumor, it is assumed that a person tells a rumor to anyone he meets until he encounters another person who has already heard it. If the population is large, it has been proved that, in the end, approximately 80% of the population will know the rumor. In this paper we will derive the (asymptotic) mean of the time it takes until the rumor dies away. It is shown that this time grows rather slowly with population size. In fact, if the population consists of W persons, and if a person meets in the mean mu persons per time unit, then the asymptotic mean duration is approximately [2.68 ln(W) + 1.34]/mu time units. PMID- 8400577 TI - An improved hybrid HIV/AIDS model geared to specific public health data and decision making. AB - An improved version of a previously described compartmental dynamic model for the spread of the HIV virus and AIDS is presented in which the estimation of key parameters depends entirely on the use of good public health data. This means that practical applications to specific regions, using only local data, can be of great value to public health decision makers dealing with local problems. It is assumed that scientific support is available within an interdisciplinary operations research context. The improved model incorporates physicians' delays in reporting AIDS incidence, additional cases revealed by death certificate analysis, and a high-risk core group of HIV positives involving a limited residence time, followed by a low-risk group with extended residence, both groups leading to AIDS cases. The model is hybrid in character in the sense that the HIV infection process with large numbers is deterministic while the incubation process with small numbers initially is stochastic. Applications have been made to Switzerland. Key parameters estimated include, in particular, the actual sizes of the original core groups for gay men and intravenous drug users. Current numbers of circulating HIV positives are also obtained. PMID- 8400578 TI - Parametric inference for epidemic models. AB - The likelihood function corresponding to epidemic data is often very complicated. We illustrate that the EM algorithm can sometimes help to simplify likelihood inferences. Difficulties with likelihood inferences about parameters of epidemic models have established a role for martingale methods. These are methods of statistical inference based on estimating equations derived from the rich theory of martingales, and they have produced simple methods of inference in a number of important applications to epidemic data. We contrast likelihood methods with martingale methods and determine which specific assumptions cause changes in inferences about the infection potential of a disease. It is found that the martingale-based estimate of the infection potential remains unaltered under a variety of commonly used model specifications but that the precision of this estimate changes as model assumptions are altered. PMID- 8400579 TI - A conditional bootstrap procedure for reconstruction of the incubation period of AIDS. AB - Data on the incubation period of AIDS patients are often fragmented and censored. parametric models have been proposed in the literature to impute the missing segment of the incubation period. The numerical results vary widely with the parametric models used. We propose a nonparametric conditional bootstrap (CB) procedure for imputation. The quality of the CB data is studied by checking the asymptotic accuracy of the CB estimators. We establish the asymptotic accuracy of the CB procedure for two basic nonparametric estimators: the empirical distribution function and a kernel-type conditional empirical distribution function. The rates of convergence of the CB approximation are obtained. The results for the kernel-type estimators hold also for the nearest-neighbor-type estimators. PMID- 8400580 TI - Estimation of vaccine efficacy from epidemics of acute infectious agents under vaccine-related heterogeneity. AB - A stochastic epidemic model is formulated for the study of the protective effects of vaccination in a population that is stratified by vaccine-related factors. The epidemic model is transformed into a counting process, and then martingale-based methods are used to provide estimators of vaccine efficacy and their variances. Following an example, various extensions of the model are discussed. PMID- 8400581 TI - A three-state Markov model of Plasmodium falciparum parasitemia. AB - A three-state Markov model taking into account clinical signs of malaria infections by P. falciparum is described. The three states considered are the noninfected (state 0), the infected exhibiting no clinical signs (state 1), and the infected with clinical signs (state 2). Methods for estimating the transition rates from longitudinal data are indicated. This model was used to assess the effect on children of an intervention trial on the use of mosquito nets impregnated with insecticide. The trial was conducted in West Africa (Burkina Faso) between 1985 and 1987. The analysis showed that the intervention was most effective on transition rates between state 1 and state 2. PMID- 8400582 TI - A threshold limit theorem for a multitype epidemic model. AB - The asymptotic final size distribution of a multitype Martin-Lof process, a discrete-time SIR model for the spread of an infectious disease in a closed multitype population, is derived for the case of large total population size. When all subgroups are of comparable size and the infection pattern is irreducible, a threshold behavior is obtained, and the asymptomatic distributions for small and large outbreaks can be found. As an important corollary we get a threshold limit theorem for a class of continuous-time SIR models with several types. PMID- 8400583 TI - The basic reproduction ratio for sexually transmitted diseases. Part 2. Effects of variable HIV infectivity. AB - In this paper we investigate the effects of variable infectivity on the spread of HIV in a heterosexual population where pair formation and separation are taken into account. We calculate the basic reproduction ratio as a function of the number of new partners during the infectious period, keeping the total number of contacts fixed. Numerical evidence suggests that the basic reproduction ratio decreases for variable infectivity if the average infectivity is kept constant. PMID- 8400584 TI - The correlation structure of epidemic models. AB - For a general Markov SIS epidemic model, the fates of individuals at different times are shown to be positively correlated. When the population is subjected to two diseases, a certain condition, here called positive interference, results in positive correlations between individuals with respect to either disease, while another condition, called competition, gives negative correlation between diseases and positive correlation within each disease. The results generalize to two classes of disease, with positive interference within each class and competition between classes. A general (non-Markov) SIR model (which includes the general epidemic and generalized Reed-Frost models) exhibits positive correlation. The results for SIS models rely heavily on monotonicity properties and in some cases on a careful choice of partial order. For the SIR models a graphical construction of the models is used. PMID- 8400585 TI - The stochastic SI model with recruitment and deaths. I. Comparison with the closed SIS model. AB - We compare the stochastic and deterministic versions of an SI model with recruitment, background deaths, and deaths due to the disease. For the stochastic version, analysis of the mean number of susceptibles, mx, and infecteds, m(y), and of the means conditioned on nonextinction of the infection, m*x and m*y, shows that (1) if R0 < or = 1, the disease dies out monotonically for the deterministic and stochastic models, and (2) if R0 > 1, the disease dies out early with a probability close to (1/R0)a, where a is the number of infecteds introduced, or m(y) rises to a peak and then dies out slowly. For small populations, N, the peak is an obvious maximum. If N > or = 100, the peak in m(y) is hidden in a long, nearly stationary plateau and m*y is close to the deterministic endemic level for a large range of parameter values. The analytical results are illustrated with simulations. The results for the SI model are motivated by and compared with the corresponding results for the closed SIS model. PMID- 8400586 TI - Role of clinical pharmacy specialist in nutrition management of bone marrow transplant patient. AB - I describe the role of the clinical pharmacy specialist (CPS) in the nutrition management of bone marrow transplant (BMT) patients. An example is used to demonstrate the complex nature of nutritional support in BMT patients and the ability of the CPS to provide expertise in the fluid and electrolyte aspect of nutrition management. This example also illustrates the multitude of factors considered by the CPS before changes in parenteral-nutrition therapy are recommended to BMT physicians. The role of the CPS in nutrition assessment, parenteral-formula design, consultation on compatibility and stability issues, formula alteration to meet changing nutrition requirements, and home nutrition management is outlined. PMID- 8400587 TI - Total parenteral nutrition in posthepatectomy patients. AB - Blood biochemical and nutritional metabolism indices were examined in eight patients who received infusion containing glucose, fructose, and xylitol in a 4:2:1 ratio (group GFX) after liver resection compared with those in six patients who received only glucose (group G). Preoperative patient-selection criteria consisted of a parabolic oral glucose tolerance test level over time, a total activity of coagulation factors II, VII, and X of > or = 60%, and an indocyanine green disappearance rate (ICG K) of > or = 0.13. Total parenteral nutrition (TPN) was started on the 3rd postoperative day. Levels of blood biochemical indices, rapid-turnover proteins, and urinary 3-methylhistidine were measured, and amino acids and nitrogen balance were analyzed preoperatively and on the 2nd, 5th, and 7th postoperative days. In group GFX, serum glutamic oxaloacetic transaminase and glutamic pyruvic transaminase levels decreased soon after TPN was begun, being 47 +/- 8 and 84 +/- 14 U/L, respectively, on the 7th postoperative day. This level was significantly lower (p < 0.05) than that in group G on the same day (94 +/- 18 and 141 +/- 22 U/L, respectively). There was no difference between the two groups in levels of rapid-turnover proteins or in Fischer ratio of amino acids. Urinary 3-methylhistidine level decreased soon after TPN in group GFX. Nitrogen balance became positive on the 7th postoperative day in group GFX, whereas it remained negative until the 7th postoperative day in group G.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400588 TI - Burn severity, copper dose, and plasma ceruloplasmin in burned children during total parenteral nutrition. AB - Copper (Cu) is an essential nutrient with known metabolic roles in wound healing. Ceruloplasmin (CP), the primary Cu-transport protein, responds as an acute-phase reactive protein after trauma. However, for severe burn trauma, this response is absent in the early catabolic phase despite Cu provision. We report data for 14 severely burned children receiving Cu in total parenteral nutrition (TPN-Cu) ranging from 7 to 26 micrograms Cu.kg-1 x day-1. All patients manifested low plasma levels of CP. The reduction in CP reflected burn severity but also appeared to be dependent on the Cu dose. Increasing Cu supplementation to improve CP raises a concern for hepatotoxicity, which is accompanied by an elevation in the plasma nonceruloplasmin Cu (nonCP-Cu). The calculated plasma nonCP-Cu in our series is consistent with a lack of increased risk and suggests that TPN-Cu at the general pediatric guideline of 20 micrograms.kg-1 x day-1 is safe and reasonable for severely burned children. Cu supplementation > 20 micrograms/kg may be beneficial; however, monitoring of both CP and total Cu should continue as standard practice in the management of these patients. PMID- 8400589 TI - Posttraumatic hormonal environment during total parenteral nutrition. AB - Hormonal responses to major trauma trigger a cascade of metabolic adjustments leading to catabolism and substrate mobilization. Energy deficit and energy surfeit have profound effects on hormone levels. To characterize the course of changes in regulatory hormone levels after multiple injury, we measured the plasma levels of eight hormones, once within 48-60 h after injury in the fasting state and then daily for 5 days during the administration of total parenteral nutrition in 10 hypermetabolic, highly catabolic, and severely injured adult patients. Acute deficiency in anabolic insulinlike growth factor 1 (IGF-1) and growth hormone levels and elevated levels of counterregulatory stress hormones and insulin were seen as a result of trauma. Provision of nutrition on the 1st day has no effect on IGF-1 and cortisol levels. However, growth hormone levels are raised to normal, and the nitrogen balance is improved. Over the next 4 days, there were no appreciable changes in these parameters. The persistent low levels of IGF-1 reflect the altered nutrition status of the patients, as characterized by the continued negative nitrogen balance and elevated cortisol levels in the early posttrauma period. Anabolic IGF-1 and insulin levels showed significant negative correlation with the catabolic indicators 3-methylhistidine and catecholamine excretion. The results suggest that IGF-1 is regulated by nutritional intake independently of growth hormone and may be a better nutrition indicator. PMID- 8400590 TI - Subjective global assessment: alternative nutrition-assessment technique for liver-transplant candidates. AB - The interpretation of most objective nutrition assessment parameters is affected by liver disease. A subjective global assessment (SGA) tool adapted for assessment of adult liver-transplant candidates was evaluated for interobserver reproducibility. Two dietitians assessed 20 liver-transplant candidates in regard to 25 SGA parameters based on patient history, physical appearance, and existing clinical conditions. Fifteen percent of the patients were found to be well nourished, 70% moderately malnourished, and 15% severely malnourished. Raters agreed on the nutrition status of the patients 80% of the time. Twelve individual parameters showed fair to good interrater reliability (kappa > or = 0.500). Muscle wasting and fat depletion were determined to be the strongest predictors of the final SGA rating (kappa = 0.737, p < 0.0001, kappa = 0.632, p < 0.0001, respectively). The assessment tool itself was determined to be reliable by the Cronbach coefficient alpha-test (0.707). Although this assessment method has limitations, the findings of this study suggest that SGA as an alternative test for assessing the nutrition status of adult liver-transplant candidates has an overall fair to good interobserver reproducibility rate. PMID- 8400591 TI - Soybean oil, blackcurrant seed oil, medium-chain triglycerides, and plasma phospholipid fatty acids of stressed patients. AB - Thirty-six adult severe head injury and cerebral stroke patients in four intensive-care units were randomized to receive one of three enteral diets for 21 days. These diets, which supplied 45% of calories from fat, differed only in lipid composition. Diet A was comprised of 100% soybean oil, diet B contained a 50:50 (wt/wt) mixture of soybean oil and medium-chain triglycerides (MCTs), and diet C contained 42.5% MCT, 50% soybean oil, and 7.5% blackcurrant seed oils. Plasma phosphatidylcholine and fatty acid composition of plasma total phospholipids were determined before initiating treatment (day 0) and weekly throughout the study. Results indicated that at the start of the study, all patients had low linoleic acid (18:2 omega 6) levels compared with healthy subjects. Emulsion A disturbed the balance between several fatty acids of the omega 6 series, as exemplified by the significant increase in 18:2 omega 6 proportions. In contrast, both emulsions B and C introduced a less-pronounced rise in 18:2 omega 6 associated for emulsion C with a significant increase in dihomo-gamma-linolenic acid (20:3 omega 6) and docosapentaenoic acid (22:5 omega 3) in plasma phospholipids. Furthermore, 18:3 omega 6 change was significantly different between groups A and C and that of 20:3 omega 6 between group A and both groups B and C. Throughout the study, arachidonic acid (20:4 omega 6) exhibited remarkable steady-state levels regardless of the diet. This study shows that providing the injured body with high amounts of 18:2 omega 6 does not lead to high levels of its upper derivatives in plasma phospholipids.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400592 TI - Preoperative nutrition assessment in liver transplantation. AB - Nutrition assessment and therapy in end-stage liver disease has become increasingly important with the advent of orthotopic liver transplantation. Reduced lean body mass, increased risk of sepsis, and altered metabolism of carbohydrates, protein, and fat are characteristic of patients with liver dysfunction. This study assesses the prevalence of protein-calorie malnutrition and the relative utility of various parameters used to define protein-calorie malnutrition in 104 patients before liver transplantation. Five subgroups were identified for analysis: primary biliary cirrhosis (PBC, n = 21), sclerosing cholangitis (SC, n = 12), chronic active hepatitis (CAH,n = 34), acute hepatitis (AH,n = 11), and other liver diseases (OD,n = 26). Clinical characteristics, anthropometric measurements, secretory protein levels, 24-h urinary creatinine and urea nitrogen, and immunological studies were assessed. Significant differences between groups were noted in age, height, weight, and percentage ideal body weight (IBW), but no differences were noted with respect to triceps skin fold (TSF) and arm muscle circumference (AMC), where uniform depletion of fat and protein stores was found. Overall percentage IBW was significantly elevated (112 +/- 20, mean +/- SD, p < 0.001), whereas TSF and AMC percentage standards were 71 +/- 33 and 89 +/- 11% (respective p < 0.001). With the < 5th percentile of TSF and AMC as markers of malnutrition, 33 and 43% of patients were malnourished, respectively. Hepatic synthetic function was impaired in all groups, with overall albumin 25 +/- 0.6 g/L, transferrin 1.60 +/- 0.66 g/L, and prothrombin 16.8 +/- 6.2 s.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400593 TI - Electromyographic activity of gastrointestinal tract of patient with short-bowel syndrome. AB - Electromyographic recordings were obtained from a patient with short-bowel syndrome. Eight recording sessions were performed in almost 3 mo, during both fasting and fed states. Slow waves were always present, and their frequencies were similar during both states. Phase III of the migrating motor complex (MMC) was not identified in any of the four recordings performed in the 1st mo after the operation. Afterward, phase III was observed in three of four recordings. Ingestion of milk caused substitution of the MMC by the fed pattern in all eight recordings. We conclude that extensive small-bowel resection causes temporary disappearance of phase III of the MMC. A change from a fasting to a fed pattern after food ingestion is normal. PMID- 8400594 TI - Acute nutrition management in insulin-dependent diabetes: presentation of an index case. AB - Diabetic patients unable to ingest nutrients orally but who have functioning gastrointestinal tracts should be nutritionally supported via enteral feeding. Provision of enteral nutrition can be problematic in terms of glycemic control. Diabetes compounded with kidney failure further complicates calorie delivery and distribution. Enteral-nutrition formulas should be carefully selected with attention to macronutrient profile, specific carbohydrate constituents, and presence or absence of dietary fiber. Modular components can be added to standard enteral feedings to alter calorie distribution while enhancing calorie provision. Selecting the method of insulin infusion (i.e., intermittent subcutaneous insulin therapy versus continuous intravenous or continuous subcutaneous infusion) for maximizing glucose control is an important step in the acute nutrition management of insulin-dependent diabetic patients. Attention to nutrition indices, fingersticks, and parameters of enteral-feeding intolerance is of utmost importance. The transition from enteral to oral nutrition will depend on adequacy of oral intake as determined by calorie counts. PMID- 8400595 TI - Strategies to counteract readjustments toward lower metabolic rates during obesity management. PMID- 8400596 TI - Influence of autonomic nervous system on nutrient-induced thermogenesis in humans. AB - Experiments that have investigated the possible influence of the sympathetic and parasympathetic nervous systems on the thermic response to intravenously and orally administered nutrients are discussed. Although two thermogenic components, obligatory and facultative thermogenesis, can be demonstrated with the hyperinsulinemic clamp technique, facultative thermogenesis is less obvious after oral administration and is probably the result of insulin-mediated sympathetic nervous stimulation. On the other hand, the parasympathetic system would appear to influence the thermic response to meal ingestion by modulating obligatory thermogenesis, i.e., the rate at which nutrients are digested, absorbed, and processed by the various tissues and organs of the body. In obese individuals, impaired activity of one or both branches of the autonomic nervous system has been observed in fasting postabsorptive conditions and, on some occasions, after meal ingestion. Autonomic nervous dysfunction may be a risk factor for obesity or associated with obesity, and its early detection could provide a means of identifying individuals at risk of becoming obese and/or diabetic so that appropriate treatment can be devised. PMID- 8400597 TI - Chickens, chemoprevention and randomized trials. PMID- 8400598 TI - Metabolic adaptation to low-energy diets: impediment in dietary treatments of obesity. PMID- 8400599 TI - Nutrition awareness: a must in foreign wars. PMID- 8400601 TI - Nutrition and public influence in the health policy process. PMID- 8400600 TI - Student's t test. PMID- 8400602 TI - An investigation into some of the effects of the state of nutrition of the mother during pregnancy and labour on the condition of the child at birth and for first few days of life. 1916. PMID- 8400603 TI - Quality assurance for bone densitometry research studies: concept and impact. AB - A concept for quality assurance (QA) in bone densitometry has been developed for clinical multicenter studies. Major elements provided by a coordinating center comprise (1) consulting services and certification of participating centers in the start-up phase of the study, (2) review of scan data acquired on QA standards for cross-calibration and longitudinal assessment of scanner stability, (3) review of selected patient data as well as of problem cases during the study, and (4) comprehensive review and correction of patient results based on QA data after conclusion of the study. Limitations of phantom-based QA data should be acknowledged. Typical problems encountered during research studies and guidelines for solutions are presented. Successful implementation of QA measures may yield substantial enhancement of statistical power. Depending on the study design and the variability of response within patient groups, improvement in precision due to QA measures may reduce the smallest detectable difference between subject groups or, alternatively, sample size by a few to more than 50%, and thus may contribute to a substantial reduction in study cost. Formulae for calculation of the magnitude of these effects are presented. To maximize the net benefit, QA efforts have to be limited to levels that assure reliability of the data at acceptable QA cost. While QA programs at individual clinical sites or for local practitioners may not need to be as extensive as for multicenter clinical trials, awareness of the potential problems and implementation of basic QA measures will help in obtaining high-quality bone densitometry results. PMID- 8400604 TI - Reduced bone formation in patients with osteoporosis associated with inflammatory bowel disease. AB - The pathophysiology of bone loss associated with inflammatory bowel disease has not been clearly defined. In this study we have performed a detailed histomorphometric analysis of iliac crest bone obtained from 19 patients with inflammatory bowel disease in whom a diagnosis of osteoporosis had been made. Eleven subjects were receiving prednisolone at the time of their biopsy. Comparison with control values demonstrated a highly significant reduction in trabecular bone area in the patient group (p < 0.001). Wall width, adjusted appositional rate and bone formation rate were all significantly reduced in the patient group (p < 0.001) and the formation period was significantly increased (p < 0.001). Resorption cavities were slightly smaller in the patient group, differences in maximum cavity depth and cavity length achieving statistical significance (p < 0.005 and p < 0.05 respectively). The mineral appositional rate was significantly reduced in the patients with inflammatory bowel disease (p < 0.001) and the mineralization lag time significantly increased (p < 0.001); however, osteoid area, perimeter and seam width were not significantly different from controls. These results demonstrate that osteoporosis associated with inflammatory bowel disease is characterized by reduced bone formation at the cellular and tissue level; the proportionately greater change in wall width than in resorption cavity depth is consistent with a negative remodelling balance. Although none of the patients had osteomalacia as defined by the criteria of increased osteoid seam width and mineralization lag time, the higher mineralization lag time in the patient group indicates a mild mineralization defect. PMID- 8400605 TI - Relationship between bone mineral density and dietary intakes in the elderly. AB - Dietary protein and/or calorie insufficiencies represent an important problem in elderly patients. The biological and clinical implications, and particularly the influence on bone mass of undernutrition in the elderly, have not been completely defined, although several studies have demonstrated a high prevalence of dietary insufficiencies in patients with a recent fracture of the proximal femur. In the present study the relationship between dietary intakes, physical performance and bone mineral density (BMD) was examined in hospitalized elderly patients. The study comprised 74 patients (48 women, mean age 82 years; and 26 men, mean age 80 years) who were hospitalized for various medical indications. They were divided into two groups according to their dietary protein intakes, evaluated during the first 28 days in hospital while on a regular diet. The first group consisted of 26 patients (14 women and 12 men) whose protein intake was equal to or greater than 1 g per kilogram of ideal body weight. The second group consisted of 48 patients (34 women and 14 men) who consumed less than 1 g of protein per kilogram of ideal body weight. The two groups differed also in their energy, carbohydrate, lipid and calcium intakes. Patients in the group with the higher protein intake displayed higher BMD at the level of the femoral neck as measured by dual-photon absorptiometry. The men in this group also had higher lumbar spine BMD. After 4 weeks in hospital the women with a higher protein intake had significantly enhanced bicipital and quadricipital muscle strength and better performance as indicated by the increased capacity to climb stairs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400606 TI - Ultrasound measurements on os calcis: precision and age-related changes in a normal female population. AB - We performed ultrasound measurements in the calcaneus of 512 healthy women. Broadband ultrasonic attenuation (BUA) and speed of sound (SOS) were obtained with a Lunar Achilles ultrasonic instrument. Subjects studied were one group of 67 women working in our hospital (group A) and two groups which are part of two large prospective cohort studies (groups B and C). Group B consisted of 244 women aged 31-79 years randomly selected from a large insurance company, and group C consisted of 201 women aged 74-91 years randomly selected from the electoral rolls. Dual-energy X-ray absorptiometry (DXA) measurements of femoral neck and total body were performed with a Hologic QDR 2000 for group B and with a Lunar DPX Plus for group C. The in vitro precision of the Achilles, estimated by measuring a phantom daily for 45 days, was 0.84% for BUA and 0.12% for SOS. We assessed the in vivo short-term precision in 20 healthy volunteers working at the hospital, measured three times each. The coefficients of variation were 0.93% (+/ 0.21) for BUA and 0.15% (+/- 0.03) for SOS. The precision error was compared with the true variation, to obtain a standardized coefficient of variation. We analysed the three groups pooled together (n = 512) and found for BUA an average 20% decrease and for SOS a 5% decrease between the ages of 20 and 90 years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400607 TI - Evaluation of bone turnover in type I osteoporosis using biochemical markers specific for both bone formation and bone resorption. AB - The aims of the study were to evaluate the use of bone-specific biochemical markers of turnover in type I osteoporosis, to test for evidence of heterogeneity of bone turnover in this condition, and to attempt to devise an 'uncoupling index' by using the relationship between bone-specific biochemical markers of bone formation and bone resorption. In women with type I osteoporosis (mean age 64 years, SD 5; n = 63) the mean level of serum osteocalcin, a specific biochemical marker of bone formation, was 9.9 ng/ml (SD 2.0), which was higher than the level in normal postmenopausal women (mean age 65 years, SD 6; n = 8.9 ng/ml (SD 2.0; p < 0.01). The variance of serum osteocalcin levels in the two groups was similar. Compared with this 11% increase in the biochemical marker for bone formation, the markers of bone resorption, total urinary deoxypyridinoline (bone-specific), pyridinoline and hydroxyproline were increased by 40% (p < 0.0001), 61% (p < 0.0001) and 25% (p < 0.01), respectively. Furthermore, these biochemical markers of bone resorption had greater variance in women in type I osteoporosis than in the normal postmenopausal women (p < 0.01). The urinary excretion of the free crosslinks deoxypyridinoline, pyridinoline and glycosylated pyridinoline were increased by 26% (p < 0.001), 17% (p < 0.01) and 13% (NS) respectively. An 'uncoupling index' was calculated for the difference between urinary deoxypyridinoline and serum osteocalcin using the results from the normal women and expressed as z-scores. We conclude that the pyridinium crosslinks of collagen enable better discrimination between normal and osteoporotic women than does hydroxyproline.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400608 TI - Prevention of osteoporosis with nasal salmon calcitonin: effect of anti-salmon calcitonin antibody formation. AB - The amino acid sequence of salmon calcitonin (SCT) differs considerably from that of the human hormone and specific antibodies (Ab) develop in a significant proportion of patients after parenteral or nasal administration of SCT. Controversy remains regarding the functional importance of these Ab. We report on the development of specific anti-SCT Ab in a population of postmenopausal women receiving nasal SCT for prevention of postmenopausal bone loss, and compare the effects of nasal SCT in women with or without Ab. Thirty-nine per cent of women developed Ab after 6 months of treatment with SCT, 52% after 12 months, and 61% after 18 and 24 months. After 24 months the AB titre was 3.47-17.7 x 10(-9) M/l (mean +/- SD: 13.3 +/- 3.1 x 10(-9) M/l). No significant differences appeared between the changes in lumbar bone mineral density (BMD) measured in the whole population (n = 44) (mean +/- SD: +1.06 +/- 3.9%), the patients without Ab (n = 17) (+0.05 +/- 3.7%) or in those with Ab (n = 27) (+1.7 +/- 4.6%). During the same period, a control population randomly assigned to a 500 mg/day calcium intake showed a significant loss of lumbar BMD (-4.57 +/- 4.9%) (p < 0.01). In conclusion, in healthy postmenopausal women nasal SCT seems to maintain the same preventive effect against bone loss whether or not Ab are present. PMID- 8400609 TI - Revisable criteria for vertebral deformity. AB - In order to study vertebral fractures in various study populations, we earlier prepared a database of vertebral dimensions derived from spinal radiographs of 191 normal women seen regularly over 25 years. In this report we have expanded the range of measurements to include vertebral levels T3 to L5. We report means and standard deviations on anterior and posterior heights, on wedge shape and on heights relative to adjacent vertebrae. When one or both of the latter two quantities are 'far' below the mean, a vertebra is called deformed. We also describe a more flexible way of expressing damage using the number of deformed vertebrae, the degree of deformity of individual vertebrae, or the total damage to the entire spine. In assessing damage we use criteria for deformity adjusted to the limits detected by an experienced diagnostician, replacing an earlier approach based on 95% probability limits of normal variation. The normal women from whom these variations are ascertained are a low-prevalence group with respect to vertebral deformity, with prevalence of 2.8%. When the criteria developed from these women were applied to a moderate-prevalence group (37%) the model had a sensitivity of 97%, a specificity of 89% and an accuracy of 92% as regards the identification of subjects with damaged vertebrae. When used epidemiologically for a moderate-prevalence group the model has a known overestimation of 15%. the model is compared with other schemes for identifying vertebral deformities. PMID- 8400610 TI - Acute biochemical variations induced by four different calcium salts in healthy male volunteers. AB - Calcium (Ca) supplements have positive effects in growing children, reduce bone loss in late-postmenopausal women with a low calcium diet and, in association with vitamin D3 supplements, may reduce non-vertebral fracture rates in elderly women. However, for many formulated pharmaceutical products their relative beneficial effects have not been conclusively established. We have compared the acute (6 h) metabolic responses following oral administration of two preparations of calcium gluconolactate and carbonate (CG and CG'), tricalcium phosphate (TCP) and calcium citrate (CC), given on separate occasions in each of 10 healthy young male volunteers. The subjects fasted overnight for 12 h and continued to fast during the experimental procedure. A 1000 mg dose of each Ca salt was ingested at weekly intervals. Blood was drawn after 30, 60, 90, 120, 180, 240, 300 and 360 min for measurement of serum Ca, phosphorus (P), parathyroid hormone (PTH) and whole plasma calcitonin (iCT). All Ca supplements induced significant (+6.4% to +8.1%; p < 0.01) increases in Ca and significant suppression of PTH (-37.4% to 57.4%; p < 0.01). Comparison of response curves revealed significantly (p < 0.01) more marked Ca increase and PTH suppression with CC than with the other three Ca salts. CG' and CC induced marginal decreases in serum P and the overall curve of P variations was different for TCP compared with CG, CG' and CC. No significant variation of iCT was recorded during the test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400611 TI - Interpretation of lumbar spine densitometry in women with fractures. AB - Identification of postmenopausal women at risk of developing osteoporotic fractures is a major clinical problem. In this study the use of projected planar lumbar bone density values for individual fracture risk assessment was questioned. Osteodensitometry (DXA) results from 415 normal women, 62 women with previous vertebral compressions, and 76 women with previous low-energy fractures were analyzed, together with their body size and lumbar vertebral body size variables. The following were found: (1) Lumbar vertebral projected bone mineral areal density (BMD) and bone mineral content (BMC) of normal women correlated with body size variables (p < 0.001). (2) Lumbar vertebral body size variables also correlated with body size variables (p < 0.001). Logistic regression analysis of measured and derived physical variables from women without and with vertebral compression fractures (n = 477) showed: (3) The best compression fracture discriminator, significantly better than BMD, was BMC divided by (Hmax/165 cm)1.5 x (D/4.35 cm)1.5, where Hmax is the body height (cm) at the menopause, and D the mean lumbar vertebral diameter of the three mid-lumbar vertebral bodies (cm). This parameter was termed BMCcorr.. ROC analysis showed: (4) At a BMCcorr. true positive ratio of 80% the corresponding uncorrected BMC or BMD true positive ratio was only 60%. The corresponding false positive ratio was 6%. Lumbar osteodensitometry could not be used to identify women with a history of peripheral low-energy fractures. (5) BMCcorr. did not, unlike BMC and BMD, correlate with body size and vertebral size variables. (6) Likewise, an observed correlation between BMC and lean body mass in a subpopulation of 116 normal women was abolished when BMCcorr. replaced BMC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400612 TI - Spurious dual-energy X-ray absorptiometry images in a patient exposed to the contrast agent Thorotrast. AB - A 69-year-old woman presented with a 20-year history of back pain and a 10 cm height loss. She had received an injection of the contrast agent, Thorotrast, at age 23. There was no history of fluoride exposure. Multiple vertebral compression fractures were seen on radiographs. Dual-energy X-ray absorptiometry (DXA) scans revealed high normal bone mineral content on the spine and, on whole body scan, visualization of the liver and spleen regions. Given the attenuation coefficient of thorium and the thorium concentrations reported for liver, spleen and vertebral bodies, it is likely that thorium was visualized in the liver and spleen and that it caused spurious elevation in her DXA bone mineral content values. PMID- 8400613 TI - Changes in calibration standards for quantitative computed tomography: recommendations for clinical practice. PMID- 8400614 TI - Two immunogenic peptide conjugates derived from P. falciparum MSA2 give rise to antibodies that crossreact with a nonhomologous 195-kDa malarial protein. AB - Merzoite Surface Antigen 2 (MSA2) is a 51-kDa antigen from P. falciparum that has marked sequence variation between malarial strains but contains highly conserved N- and C-terminal sequences. Previously described peptide sequences from these conserved domains are capable, when coupled to the carrier protein diphtheria toxoid (DT), of raising antibodies that recognize MSA2 in a number of malarial isolates. In this study we report the existence of two peptide conjugates from the conserved C-terminus of MSA2 which, when used as immunogens, produce antibodies that react with a 195-kDa protein and give a merzoite surface pattern on immunofluorescence assay (IFA). The antisera against these peptide conjugates do not recognize MSA2; however, the 195-kDa protein, and identical surface pattern on IFA, is recognized by a monoclonal antibody (mAB) against Merzoite Surface Antigen 1 (MSA1). These two peptides, when unconjugated to DT, are nonimmunogenic, and DT alone is not capable of inducing antibodies that react with malarial proteins. The combination of the carrier and either peptide is capable, however, of raising antibodies against MSA1, despite sharing no sequence homology with this antigen. This data thus highlights the importance of protein conformation when generating antibody against carrier-peptide conjugates. PMID- 8400615 TI - Synthesis and conformational analysis of (alpha Me)Leu/Aib model peptides. AB - We have synthesized by solution methods and fully characterized a variety of (alpha Me)Leu/Aib model peptides to the octapeptide level. A solution conformational analysis was performed by using infrared absorption. 1H nuclear magnetic resonance, and circular dichroism. The crystal-state structures of Z-D (alpha Me)Leu-(Aib)2-OtBu, pBrBz-(Aib)2-D-(alpha Me)Leu-(Aib)2-OtBu, and Ac (Aib)2-D-(alpha Me)Leu-(Aib)2-OtBu monohydrate were solved by x-ray diffraction. The results indicate that the (alpha Me)Leu residue may be easily incorporated into beta-bends and 3(10)-helical structures, and suggest that this residue tends to induce a helix handedness opposite to that promoted by its unmethylated counterpart (Leu) of the same optical configuration. PMID- 8400616 TI - Chemical synthesis of an O-phosphorylated tyrosine analogue of human angiotensin II, [Tyr(P)4] angiotensin II, and its vasoconstrictor effect in intact sheep. AB - The derivative N alpha-fluorenylmethoxy-carbonyl-O-dimethylphosphono-L-tyrosi ne was utilized in continuous-flow solid-phase synthesis to prepare the O phosphotyrosine analogue of human angiotensin II, [Tyr(P)4] angiotensin II. Side chain deprotection, including the removal of the methyl phosphate groups, as well as cleavage of the peptide from the solid support was achieved with 1 M trimethylsilyl-bromide-thioanisole in trifluoroacetic acid. Overall yield of purified peptide was 46%. The pressor response in intact sheep to graded infusions of the synthetic [Tyr(P)4] angiotensin II showed it to have similar potency to the native angiotensin II. However, a prolonged, elevated mean arterial pressure was observed following cessation of the infusion. PMID- 8400617 TI - Secondary structure of a human growth hormone-releasing factor fragment (Leu27 hGRF(15-32)NH2) in aqueous/SDS micelle environments. AB - The secondary structure of a human growth hormone-releasing factor (hGRF) fragment (Leu27-hGRF(15-32)NH2) has been studied by 1H NMR at 500 MHz in aqueous solutions containing varying concentrations of d25-sodium dodecyl sulfate (SDS). Chemical shifts, coupling constants and NOESY data show that the secondary structure of the peptide is random in aqueous solution in the absence of SDS. At relatively low molar ratios of SDS to peptide (1.3:1 to 3.3:1 SDS:peptide) the 1D 1H spectrum of the peptide changes as the peptide resonances are broadened significantly. NOESY patterns consistent with helical structure are present in the region of residues 22-29 when the SDS:peptide molar ratio is 1.3:1 and the SDS concentration is slightly below the critical micelle concentration (CMC). At higher molar ratios of SDS to peptide (16:1 to 72:1), where the SDS concentration is significantly above the CMC, the lineshape of the peptide's 1H NMR spectrum is sharpened. In these environments an alpha-helical conformation is induced in residues 19-32 of the hGRF fragment, as shown both by NOESY and by chemical shift data. Thus, the well-known tendency of this region of the GRF peptide to form alpha-helix in isotropic mixed-solvent systems (e.g., methanol/water, trifluoroethanol (TFE)/water) is seen also in SDS/aqueous systems. PMID- 8400618 TI - Development of a fully automated multichannel peptide synthesizer with integrated TFA cleavage capability. AB - A fully automated multichannel peptide synthesizer has been constructed which performs simultaneous and rapid assembly of peptides on a 20-200 mumol scale. In situ activation of amino acids using BOP or PyBOP was chosen to give an optimized coupling chemistry. Specially designed blocks of valves, with a zero dead volume combined with an original circuitry, permit the distribution of amino acids derivatives and reagents pre-dissolved in DMF. Either Boc or Fmoc chemistry can be adapted on the synthesizer. In Boc synthesis a very rapid protocol involving Boc group deprotection in neat TFA, followed by the concomitant steps of neutralization and coupling, allows the addition of three amino acids per hour on each channel. In Fmoc chemistry we have integrated into the synthesizer an automatic TFA cleavage system that allows the peptides to be cleaved from the resin directly within the reactors used for synthesis. The stability of the Fmoc amino acid derivatives in solution in DMF was investigated, and decomposition was found to be insignificant during the time-span of a synthesis. The satisfactory performance of the instrument was demonstrated by routine synthesis of 15-20 mer peptides. PMID- 8400619 TI - Risk assessment uncertainties. PMID- 8400620 TI - Glutathione depletion potentiates ethyl methanesulfonate-induced damage to sperm chromatin structure. AB - Male rats were treated with phorone at dosages previously shown to reduce glutathione in rodent reproductive tracts, followed by a single challenge with ethyl methanesulfonate, a known mutagenic and clastogenic agent. Epididymal sperm collected 8 and 15 days after exposure from phorone pretreated animals had a significantly greater alteration of sperm chromatin structure, defined as an increased susceptibility to DNA denaturation in situ, relative to sperm obtained from animals injected with saline alone or saline+EMS (50, 100, 150, or 200 mg/kg bw). These data support the hypothesis that ethyl methanesulfonate-induced alkylation of developing sperm chromatin protamines causes a significant stress on chromatin structure leading to increased DNA damage. This is the first report showing that glutathione depletion potentiates EMS-induced chromatin structural alterations that are likely related to dominant lethal mutations. PMID- 8400621 TI - Testicular toxicity of boric acid (BA): relationship of dose to lesion development and recovery in the F344 rat. AB - High-dose boric acid (BA) produces testicular lesions in adult rats, characterized by inhibited spermiation followed by atrophy. The present study addressed whether inhibited spermiation can be separated from atrophy based on dose, compared testis boron (B) dosimetry to lesion development, determined how inhibited spermiation was reflected by common reproductive endpoints, and examined reversibility of the testicular lesions. Rats were fed 3000, 4500, 6000, or 9000 ppm BA for up to 9 weeks and examined. Recovery was assessed for up to 32 weeks post treatment. Inhibited spermiation could be separated from atrophy based on dose (inhibited spermiation: 3000/4500 ppm; atrophy: 6000/9000 ppm), with each lesion aspect expressed at different threshold testis B concentrations (inhibited spermiation: 5.6 micrograms B/g and atrophy: 11.9 micrograms B/g) with no B accumulation during the 9-week exposure. These data suggest that separate mechanisms may be operating for these lesion aspects based on testis B concentration and that B dose rate was important for testicular toxicity. Inhibited spermiation was most reliably reflected by informed testicular histology, with the more severe cases decreasing epididymal sperm count to levels that could affect fertility. After treatment, serum and testis B levels in all dose groups rapidly fell to background levels at the earliest time points evaluated (7 days and 8 weeks posttreatment, respectively). The severely inhibited spermiation at 4500 ppm was resolved by 16 weeks posttreatment, but areas of focal atrophy were detected that did not recover posttreatment. Also, no signs of recovery from atrophy were observed (6000 and 9000 ppm). Atrophic tubules contained a normal complement of spermatogonia (2.6 to 2.9 germ cells/100 Sertoli cells), with occasional dividing and degenerating germ cells. Elevations in serum FSH and LH levels suggested an intact hormonal response to the atrophy. In summary, 1) the different aspects of the BA-induced testicular lesion can be separated using different doses, 2) inhibited spermiation does not necessarily proceed to atrophy, and 3) there is no recovery from the atrophy despite the absence of testis B after treatment. The ability to separate inhibited spermiation from atrophy based on dose and testis B dosimetry will be useful in evaluating possible mechanisms. Furthermore, the presence of dividing spermatogonia during long-term BA-induced atrophy suggests that this model should be useful for identifying critical components involved in the reinitiation of spermatogenesis. PMID- 8400622 TI - Role of laminin autoantibodies on the embryo toxicity of sera from mercuric chloride treated brown Norway rats. AB - In previous studies, antilaminin antibodies were found to be toxic to cultured rat embryos. In order to extend these studies, Brown Norway rats were treated with mercuric chloride, which led to the production of laminin autoantibodies. Sera samples from brown Norway rats treated with mercuric chloride were found to be teratogenic as well as lethal to cultured rat embryos. This embryotoxicity was not associated with sera mercury levels, but was related to the levels of antilaminin antibodies in sera. Affinity purified laminin antibodies from these mercuric chloride treated Brown Norway rats, when added to control sera, were found to be teratogenic but not lethal. These antibodies were found to bind to the laminin sequences IKVAV (A chain) and YIGSR (B1 chain), but not RGD (A chain) or YD (B1 chain). These observations suggested the possibility that an environmental pollutant such as mercury could cause the formation of embryotoxic autoantibodies that could persist in the body as embryotoxic factors for extended periods of time. PMID- 8400623 TI - Temporal evolution of risk estimates for presumed human teratogens. AB - We present preliminary data assessing a previously untried method of deriving estimates of risk from case reports on presumed human teratogens. We postulated that we could take advantage of biases inherent to case reports in order to generate one or more families of temporal curves that could be used to estimate the "true" risk of teratogenic exposure. Using this method (which we refer to as the "case-cumulative method") we found that two agents (parvovirus B19 and isotretinoin) demonstrated a logarithmic decrease in the estimated risk over time, as intuitively expected, while trimethadione and the coumarin derivatives showed a more complex pattern over time. Analysis of estimated risks quoted by reviews and large studies for these four agents showed large variability from estimate to estimate and no discernible temporal pattern. With further analysis of other agents, the case-cumulative method might eventually prove to be useful in teratogen counseling. PMID- 8400624 TI - Altered steroidogenesis in whole-ovary and adrenal culture in cycling rats. AB - Cultures of minced, whole-ovary (whole-ovary culture) were used to determine if three selected chemicals altered steroidogenic profiles. First, phenolsulfonthalein (PST), when used in culture medium, was tested for its influence on in vitro steroidogenesis. Next, aminoglutethimide (AGTP; 0 or 150 mg/kg once) and di(2-ethylhexyl)phthalate (DEHP; 0 or 1500 mg/kg/day for 10 days) were administered in vivo to young adult cycling rats, and the ovaries and adrenals were removed and cultured for 1 h. Ovarian steroidogenic profiles of progesterone (P), testosterone (T), and estradiol (E) release into the medium were measured using radioimmunoassay techniques. PST in medium significantly decreased ovarian P production and altered T and E production so that the T/E ratio was significantly altered. Therefore, PST was excluded in the later studies. DEHP altered steroid profiles so that proestrus appeared to be delayed. AGTP decreased P and E production significantly, and T production was increased slightly in proestrus ovaries. These AGTP alterations in T and E resulted in a highly significant increase in the T/E ratio. Adrenals from the DEHP and AGTP experiments were also cultured for 1 h, and P was assayed in the medium. AGTP, but not DEHP, significantly increased the production of P in adrenals. Whole ovary culture is recommended as an in vitro test for chemicals suspected of interfering with steroidogenesis in vivo. This test model should be placed strategically between in vivo studies of reproductive toxicity and complex in vitro mechanistic studies. PMID- 8400625 TI - Assessment of the effectiveness of animal developmental toxicity testing for human safety. AB - Evaluations of studies for four well-known human developmental toxicants clearly suggest that a margin of exposure of 1/100th the NOAEL for the most sensitive animal species tested provides adequate safety for the human conceptus. The lowest reported human teratogenic exposures occurred at doses at least one log above the estimated "safe" or acceptable daily exposure based on the most sensitive animal species, that is, 1/100th animal NOAEL. (The MOE ranged from < 1 to 10.). The data and analyses are consistent with the conclusion that, regardless of the type of in utero effect produced in animals, the margin of safety of 100 is likely to protect the human conceptus in utero from developmental perturbation, and it is a scientifically reasonable and conservative default number. PMID- 8400626 TI - Confidence intervals. PMID- 8400627 TI - Evaluation of brompheniramine safety in pregnancy. PMID- 8400628 TI - In vitro tests for teratogens: desirable endpoints, test batteries and current status of the micromass teratogen test. AB - Information from in vitro tests can be usefully used as a component of the risk/hazard assessment process. In vivo studies will be required to confirm the in vitro data. If the in vitro test system is designed around endpoints that reflect changes following in vivo toxic insult then it may be possible to modify the in vitro system to account for some of the discrepancies observed between in vivo and in vitro outcomes. When the discrepancy can be accounted for by low bioavailability in vivo, pharmacokinetic studies may be required to determine the relevance of the in vitro toxic concentrations. Reproductive hazard, especially teratogenicity, has been the subject of intensive in vitro test development. The observation of teratogenicity may affect the development of new products more significantly than any other type or category of reproductive toxicity. The micromass test, involving culture of differentiating rat embryo limb and midbrain cells exposed to test agents, may be useful as part of a battery of in vitro tests for teratogens. The most recent protocol for the micromass test is described, followed by a summary of validation and mechanistic studies confirming its usefulness. The test is robust in its transfer to new laboratories. Interlaboratory variability is small. PMID- 8400629 TI - Structure-activity approaches in the screening of environmental agents for developmental toxicity. PMID- 8400630 TI - Test guideline development and animal welfare: regulatory acceptance of in vitro studies. AB - Toxicity studies are necessary in order to be able to identify the potential hazards of chemical interference with human reproduction. Until today, most useful contributions to the assessment of possible human reproductive toxicity are considered to be made by animal studies. The OECD Test Guidelines provide the international standards for safety testing and, consequently, have traditionally focused on animal studies. However, the OECD Member countries consider the welfare of laboratory animals also of importance and are of the opinion that animal welfare considerations should significantly influence the work in the OECD Chemicals Programme. A constant effort is being made to discover alternative testing systems and to achieve their regulatory acceptance. However, activities predominately focus on finding alternatives to existing animal studies, rather than developing nonanimal tests that could contribute significantly to the hazard identification process. An approach based on the selection of endpoints essential for hazard identification that would focus on demonstrating similarities with the "real life" target events is considered likely to achieve regulatory acceptance much earlier than an approach based on high correlations between the alternative method and the existing animal study it is supposed to replace, simply because it is never better than the existing method, but at most "almost as reliable." PMID- 8400631 TI - In vitro approaches in developmental toxicology. AB - While the usefulness of in vitro screens in developmental toxicology seemed obvious a decade or more ago, integration of in vitro or short-term screens for developmental toxicity into the toxicology armamentarium of tests has been very slow. The large number and complexity of critical events in normal development in vivo limit the usefulness of in vitro systems that measure a very narrow range of developmental events, even though such systems are very useful for mechanistic research. Better understanding of the regulatory mechanisms involved in normal development that is evolving from molecular and developmental biology laboratories should help to identify critical events and processes in development that could serve as functional screens for abnormal development in the future. PMID- 8400632 TI - Significance of embryo culture methods for studying the prenatal toxicity of virustatic agents. AB - Culture methods have become important tools for elucidating the prenatal toxicity of drugs and other xenobiotics. In this paper we will review how we used in vitro as well as in vivo approaches to demonstrate the teratogenic potential of aciclovir and other related virustatic agents. In addition, some new data on this topic will be given. The teratogenic potential of the virustatic agent aciclovir was not recognized in routinely performed segment-II-studies, but the first indication came from experiments with the culture of rat embryos. Subsequently, the findings were confirmed in modified in vivo tests and it became clear that out of a group of six related drugs aciclovir exhibited the highest potential for prenatal toxicity. The effects of aciclovir on limb development were not pronounced--this has been shown with in vitro and in vivo experiments as well. In vivo experiments first indicated that the prenatal development of the thymus is disturbed by aciclovir. This effect was further studied with the culture of fetal thymuses and again the effect of aciclovir could be compared with related drugs. In summary, our work with virustatics during the last years has shown that in vivo and in vitro approaches are by no means competitive and that a combination of both approaches can provide a solid basis for a toxicologic evaluation. PMID- 8400633 TI - Regulation of growth and differentiation in early development: of mice and models. AB - In this article we describe some of the fundamental processes occurring during early murine development, introduce cellular models used to investigate these processes and review some well-known factors that may be involved in their control. These include transforming growth factor beta, retinoic acid and leukaemia inhibitory factor. Refinements to the culture conditions of embryonic stem and embryonal carcinoma cells have enabled us to test the effects of these factors on growth and differentiation and in particular to establish that their interaction may determine the ultimate developmental state of the cell population. Preliminary studies using neutralizing antibodies in embryos are described that suggest that deregulation of normal expression can lead to a failure to implant. Insights into the events underlying normal embryonic development and implantation, yielded by the type of study described here, may contribute to an understanding of the mechanisms causing early embryonic loss and the role of toxicants in this process. PMID- 8400634 TI - Embryonic and fetal development: fundamental research. AB - Much progress has been made over the past decades in the development of in vitro techniques for the assessment of chemically induced effects in embryonic and fetal development. In vitro assays have originally been developed to provide information on the mechanism of action of normal development, and have hence more adequately been used in fundamental research. These assays had to undergo extensive modification to be used in developmental toxicity testing. The present paper focuses on the rat whole embryo culture system, but also reviews modifications that were undertaken for the in vitro chick embryo system and the aggregate cultures of fetal rat brain cells. Today these tests cannot replace the existing in vivo developmental toxicity tests. They can, however, be used to screen chemicals for further development or further testing. In addition, these in vitro tests provide valuable information on the mechanisms of developmental toxicity and help to understand the relevancy of findings for humans. In vitro systems, combined with selected in vivo testing and pharmacokinetic investigations in animals and humans, can thus provide essential information for human risk assessment. PMID- 8400635 TI - Neurodevelopmental toxicity in vitro: primary cell culture models for screening and risk assessment. AB - Robust models for the evaluation of developmental toxicity are briefly reviewed with emphasis on embryonic brain and retina cells in vitro. Organ slice and aggregate cultures under constant gyratory movement as well as high cell density monolayer ("micromass") cultures are considered as robust models. An in vitro model using high cell density monolayer and re-aggregated cells isolated from embryonic chick brain (ED 6) is presented. Cell development and differentiation of the astrocytes and nerve cells are monitored by marker proteins and cytotoxicity was quantified by neutral red uptake and protein content. Four human teratogens, six possible human teratogens and six unlikely human teratogens were tested in brain and retina cells for their cytotoxic and morphologic effect. All 16 substances were classified correctly except the neurotoxicants MPTP and MPP+, both of which are strong dopaminergic toxicants in vitro as well as in humans and are therefore proposed to be classified as human neuroteratogens. Preliminary data on the lowest effect levels of four potential neurotoxicants (cadmium chloride, Ara-C, Phenytoin, MPTP) in chick brain aggregate cultures correlate surprisingly well with known toxic human plasma levels. Further validation has to be undertaken to confirm these promising results. A battery of such robust in vitro models is proposed that could cover neurodevelopmental toxicity of drugs and chemicals for screening and risk assessment purposes. PMID- 8400636 TI - Testicular and germ cell toxicity: in vitro approaches. AB - Research on testicular toxicology has been advanced significantly by the introduction of in vitro testing systems. In vivo systems, however, are still essential parts of the risk assessment process, and they are unlikely to be eliminated by in vitro model systems. While in vivo systems are needed to study the integrated male reproductive system, in vitro systems are uniquely suited to investigate specific mechanisms of action in the testis. In vitro systems substantially improve the interpretation and use of in vivo systems. In vitro models can be used alone or in combination with each other to test hypotheses about testicular toxicity. Numerous systems are described in the literature, including Sertoli-germ cell cocultures, Sertoli cell-enriched cultures, germ cell enriched cultures, Leydig cell cultures, and Leydig-Sertoli cell cocultures. These systems have been used to test relative toxicologic activity of selected chemicals in a class, to investigate the cellular response to certain toxicants, to study the metabolic capability of cells, and to describe the interaction of adjacent cell types. PMID- 8400637 TI - Sensitivity of Sertoli and Leydig cells to xenobiotics in in vitro models. AB - Different chemicals are known to cause testicular damage in the human male and experimental animals. However, the ability to assess the potential and mechanism of action leading to chemically-induced damage in men has been hampered by a lack of good predictive models. Although many of these chemicals were found to impair reproductive capacity in various laboratory animals, only some have caused reproductive damage in men. Mammalian spermatogenesis takes places within the avascular seminiferous tubules of the testis. Specialized tight junctions, which form between adjacent Sertoli cells at the time of puberty, divide the tubular space into the basal and adluminal compartments, and create a "blood-testis" barrier that restricts passage of substances and ions from the circulation. Thus, the completion of meiosis and post-meiotic germ cell differentiation, which take place in the adluminal compartment, are isolated from circulating substances unable to cross the blood-testis barrier. It seems feasible, therefore, that damage to the germ cells induced by testicular toxicants may be mediated through other cells in the testis such as the Sertoli, peritubular, or Leydig cells. A recently developed two-compartment system for culture of testicular cells can simulate, to some degree, the normal physiologic conditions. In principle, Sertoli cells isolated from mammalian testes are cultured on a permeable support (that is millipore filter) between two fluid compartments. They form a highly polarized epithelial layer with characteristic tight junctions that restrict the passage of substances between the two compartments, in analogy to the blood testis barrier. We believe this system provides an excellent in vitro model for determining the ability of chemicals to: a) alter the permeability of the blood testis barrier, b) impair the secretory function of Sertoli cells, or c) affect their viability, all of which could indirectly affect the germ cells. We have utilized this system for examining the effects of cadmium chloride (CdCl2) and other toxic substances known to affect the testis. The Leydig cell toxicity was investigated in testicular perfusion system or cultures of isolated Leydig cells. PMID- 8400638 TI - Methods and concepts in detecting abnormal reproductive outcomes of paternal origin. AB - There is conclusive evidence that exposures of human males to ionizing radiation or certain chemicals can diminish sperm production and reduce fertility. Of approximately 100 chemical agents and mixtures that have been evaluated in men by semen analysis, about half (mostly drugs and a few occupational exposures) reduced sperm quantity and quality; several of these agents also affected the fertility of exposed men. It is now well recognized that the importance of the father in reproduction goes beyond fertilization. Abnormalities in paternal chromosomes (structural and numeric) have been found in various abnormal reproductive outcomes, including chromosomal abnormality syndromes among newborns. In rodent systems, exposure of males to mutagens before mating induces transmissible cytogenetic and genetic abnormalities as well as morphologic defects and cancer among offspring. Consistent with animal findings, there is growing epidemiologic evidence of associations between male exposures to exogenous agents and abnormal reproductive outcomes (fetal loss, birth defects, childhood cancer, etc.). However, no clear links have been established between exposure, mechanism of transmission, and abnormal reproductive outcomes. It is not known to what extent male-mediated birth defects and childhood cancer are due to genetic, epigenetic, or nongenetic causes. Viewed in a multigenerational context, the role of the father in abnormal reproductive outcomes is dependent on his exposure history and susceptibilities as well as those of his mate. Relevant exposures may occur any time between conception of the parents and production of their fertilizing gametes, including their development in utero, childhood, and adolescence. Efficient measurements (including biomarkers) of relevant exposure, early biologic effects, and susceptibility in human males are under development. An integrated approach is recommended for assessing male reproductive and genetic toxicity that utilizes biomarkers in (a) epidemiologic studies of exposed human populations, (b) risk characterization in sensitive laboratory species, and (c) in vivo and in vitro studies of the molecular mechanisms of action of toxicants. A special category of "bridging" biomarkers is needed for evaluating animal data for risk assessment and for discriminating among genetic, epigenetic, and nongenetic mechanisms of abnormal reproductive outcomes of paternal origin. PMID- 8400639 TI - Methods to evaluate reproductive health of the human male. AB - The proportion of men suffering from subfertility has increased and sperm quality of the general population has decreased rather dramatically during the last decades. Conventional methods to measure sperm concentration, motility, and morphology give a rough estimate of the fertilizing capacity of semen. Objective assessment of motility characteristics and morphometric image analysis may give more precise information, but technology still needs to be refined. Some aspects of the functional capacity of spermatozoa may be impaired before measurable changes occur in their number or morphology. A battery of tests is described to evaluate sperm migration and capacitation, acrosome reaction and acrosin activity, zona adhesion and binding, sperm oolemma fusion, sperm head decondensation, and chromatin make-up. Also methods are described to localize the possible site of impairment of the genital tract. Implementation of these methods may enhance our capability to detect minor effects of environmental hazards on male reproductive health. PMID- 8400640 TI - Detection of genetic damage in human sperm. AB - Assessment of genetic damage in human sperm is now possible using the hamster oocyte/human sperm fusion technique. This cross-species in vitro assay provides precise human sperm karyotypes that can be banded for identification of individual chromosomes and numeric and structural abnormalities. This technique is time-consuming and difficult but there are now 11 laboratories in 7 different countries reporting results on normal men and men at increased risk of sperm chromosomal abnormalities. Men who have been exposed to radiotherapy and chemotherapy have been shown to have an increased frequency of chromosomal abnormalities in their sperm. A few laboratories have reported results on the use of fluorescence in situ hybridization (FISH) of interphase sperm nuclei. This appears to be a promising new technique for the assessment of aneuploidy and FISH results from our laboratory correlate well with our frequency of aneuploidy determined by sperm chromosome analysis. Thus it is possible that, in the future, FISH might be used as a simple rapid screen of potentially aneugenic agents. PMID- 8400641 TI - Sites of female reproductive vulnerability: implications for testing and risk assessment. AB - All reproductive toxicity testing, whether in vivo or in vitro, should be conducted with consideration of the ultimate use of the data for reproductive risk assessment and the protection of human reproductive health. In this review selected sites of vulnerability in the female reproductive system are identified. These sites of female reproductive vulnerability may be utilized for in vitro toxicity testing assays, and the data from assays may be employed in the hazard identification and hazard characterization steps necessary for reproductive risk assessment. Using biomarkers of female reproductive function derived from in vitro toxicity testing it is possible to define functions that characterize female fecundity. These characterizations of female fecundity may be used to quantitate reproductive risk in human populations. PMID- 8400642 TI - Toxicity testing using the isolated in vitro perfused ovary. AB - This article discusses the use of in vitro perfusion techniques as a tool for toxicity testing in the ovary and how the rat ovary has been adapted for this purpose. A brief review of the development of in vitro ovarian perfusion is provided, focusing on steroidogenesis and physiology of ovulation. Adaptation of this model for use as a toxicologic model is discussed in the context of other isolated organ models, (that is, liver, heart, lung). Surgical procedures, perfusate and criteria for viability are outlined. Advantages of this technique are highlighted including ability to administer high doses of drugs directly to intact organ devoid of other influences. Applications of this model are discussed and data from studies of glutathione depleted ovaries perfused with hexachlorobenzene (HCB) are presented. Increased oxygen consumption after addition of HCB is suggestive of a disordered respiratory metabolism and is an example of future markers of ovarian injury using this innovative technique. PMID- 8400643 TI - In vitro approach to fertility research: genotoxicity tests on primordial germ cells and embryonic stem cells. AB - In vitro screening tests for reproductive toxicology are required by the 7th amendment of the directive 67/548 EEC and the OECD-programme on existing chemicals. Unfortunately, appropriate methods for testing developmental toxicity or impairment of fertility are not at hand. Therefore, we have tried to design test method based on mutagenic effects in germ cells that may be used as a test of fertility impairment as well. Embryonic stem cells (ESC) derived from mouse blastocysts can be kept in culture routinely. Establishment of ESC and improvement of their culture conditions are described and special properties of ESC are discussed in relation to germ cells. Because some properties of germ cells and pluripotent embryonic stem cells (ESC) are found to be comparable, permanent lines of ESC hold promise to be used as an in vitro test of impairment of fertility. PMID- 8400644 TI - Human trophoblast cultures: models for implantation and peri-implantation toxicology. AB - Implantation is the process that leads from blastocyst attachment to its embedding in the uterine wall. It is widely believed that failure of implantation is a common cause of pregnancy loss. Toxic agents can interfere directly with the process of implantation and therefore may account for unexplained implantation failures. Our knowledge of human implantation remains limited, mainly due to the lack of adequate experimental models. Studies of mechanisms underlying implantation in humans are by nature and for ethical reasons restricted to in vitro models. The aim of this review is to provide a critical evaluation of various in vitro models of implantation in humans, as well as essential background knowledge required for application of these models to the assessment of peri-implantation toxicity. Particular attention has been devoted to cell-cell and cell-matrix interactions as possible endpoints in the screening of toxic agents. PMID- 8400645 TI - Introduction to (pre)screening methods. AB - In summation I start this session with the opinion that in vitro methods cannot be considered as adequate replacements for entire animals at the level of regulatory testing. But, when used to identify mechanisms of action, they can be extremely useful as secondary stage supporting studies. They are of doubtful value for general purpose, broad spectrum screening of single chemical entities or for priority selection of unrelated chemicals. They can be of value for priority selection of homologous series with a known, specific effect on reproduction or development. Such situations are most likely to be present in chemical and drug manufacturing industries where judicious use of in vitro methods in an integrated approach could reduce the number of failures at the later stage of full scale testing. Whether I will need to revise my opinions at the end of this session will depend upon what our speakers have to offer. PMID- 8400646 TI - Is there any place for nonmammalian in vitro tests? AB - In vitro developmental toxicity testing procedures have two potential uses: 1) prediction of Adult/Developmental Ratios (A/D Ratios); and 2) study of the mechanisms of embryo development. Historically there has been an incorrect emphasis on malformation as the most relevant endpoint of developmental toxicity, as well as an incorrect assumption that this endpoint alone could be identified by an in vitro procedure. The literature is filled with tests that have later been shown to be inaccurate predictors of mammalian developmental toxicity. In addition, industrial experience with in vitro tests used to predict bimodal responses (e.g., potential carcinogenicity) has produced a basic mistrust of the results of these tests and an unwillingness to perform a test because of fear of inappropriate delays in the regulatory process. Because no single test has been identified as one that can replace mammalian testing procedures, there is also little need seen for these procedures, although some tests are used, not necessarily correctly, to prioritize testing. Better understanding of the utility of these tests would increase their use in the prioritizing process, possibly allow their use when remarkable acute environmental hazards occur, and assist in identifying the mechanism underlying the observed effect. PMID- 8400647 TI - Changes of the prevalence of endemic fluorosis after changing water sources in two villages in Guangdong, China. PMID- 8400648 TI - Evaluation of the biological threshold value of urinary cadmium concentration in a group of workers. PMID- 8400649 TI - Neurotoxic effects of lead exposure among printing press workers. PMID- 8400650 TI - Biological monitoring of exposure to low dose aniline, p-aminophenol, and acetaminophen. PMID- 8400651 TI - Organic solvent levels in model and hobby glues. PMID- 8400652 TI - Survey of fish and shellfish consumption by residents of the greater New Orleans area. PMID- 8400653 TI - Polycyclic aromatic hydrocarbons in seafood from the Gulf of Alaska following a major crude oil spill. PMID- 8400654 TI - Pesticide risk assessment and non-target invertebrates: integrating population depletion, population recovery, and experimental design. PMID- 8400655 TI - Natural occurrence of Fusarium toxins in barley grown in a southwestern area of Germany. PMID- 8400656 TI - Use of the bioluminescent bacterium Photobacterium phosphoreum to detect potentially biohazardous materials in water. PMID- 8400657 TI - Mutagenicity of drinking well water. PMID- 8400658 TI - Response of zooplankton to rotenone in a small pond. PMID- 8400659 TI - Toxicity of diquat pulse exposure to tropical freshwater shrimp (Caridina nilotica, atyidae). PMID- 8400660 TI - Effect of chromium and cadmium on the thermal tolerance of the prawn Macrobrachium rosenbergii exposed to hard and soft water. PMID- 8400661 TI - Effect of mercuric chloride on fertilization and larval development in the river frog, Rana heckscheri (Wright) (Anura: Ranidae). PMID- 8400662 TI - Sediment toxicity of the Anacostia River estuary, Washington, DC. PMID- 8400663 TI - Enzyme immunoassay microtiter plate response to atrazine and metolachlor in potentially interfering matrices. PMID- 8400664 TI - Fate of jet fuel JP-4 in soil. PMID- 8400665 TI - Concentrations of metals in tissues of lowbush blueberry (Vaccinium angustifolium) near a copper-nickel smelter at Sudbury, Ontario, Canada: a factor analytic approach. PMID- 8400666 TI - Effects of thiram and terbuthylazine on cellulose decomposition in two soils. PMID- 8400667 TI - Tissue distribution of cadmium-109 after tracheal and gastric administration in rats. PMID- 8400668 TI - Developmental toxicity of haloxyfop ethoxyethyl ester in the rat. PMID- 8400669 TI - Natural killer activity and serum immunosuppressive acidic protein levels in esophageal and gastric cancers. AB - The natural killer (NK) activity of peripheral blood mononuclear cells and serum immunosuppressive acidic protein (IAP) levels were examined in patients with esophageal or gastric cancer, before and after surgery. Patients with stage IV esophageal or stage IV gastric cancer had significantly lower NK activity (39.5 +/- 14.8% and 37 +/- 11.6%, respectively), and also higher serum IAP levels (778 +/- 264 micrograms/mL and 633 +/- 156 micrograms/mL, respectively), than the corresponding control values (50 +/- 5.6% and 375 +/- 26 micrograms/mL, respectively). Patients with esophageal or gastric cancer who underwent curative resection had high NK activity (54.8 +/- 11.6% and 54.8 +/- 8.0%, respectively), and low IAP levels (471 +/- 116 micrograms/mL and 490 +/- 42 micrograms/mL, respectively), compared with those who underwent non-curative resection. Patients who underwent non-curative resection had lower NK activity and higher serum IAP levels than those who underwent curative resection, even 1 month after surgery. Mononuclear cells in the regional lymph nodes and tumor specimens showed significantly lower NK activity than those in the peripheral blood and spleen. Thus, NK activity and the IAP level reflected the immunocompetence, clinical course, and surgical curability of those patients. NK cells appeared not to have any significant antitumor activity in the regional lymph nodes or in the tumor itself, although they were still active in the peripheral blood. PMID- 8400670 TI - The indications for tumor mass reduction surgery and subsequent multidisciplinary treatments in stage IV hepatocellular carcinoma. AB - The indications for tumor-mass reduction surgery and subsequent immunotherapy in patients with stage IV hepatocellular carcinoma (HCC) were elucidated in this study. About 42% of the resected specimens from stage IV-A patients (n = 26) contained well-differentiated multicentrically occurring HCC, which was not found in any of the stage IV-B patients (n = 9). The 2-year survival rate after reduction surgery was 49% for the stage IV-A patients and only 13% for the stage IV-B patients, while 6 of the stage IV-A patients who survived for more than 2 years had no vascular invasion or distant organ metastases. Some of the stage IV patients maintained normal peripheral natural killer (NK) activity and were also able to tolerate surgical insults immunologically, provided that appropriate postoperative immunotherapy was given. Thus, stage IV-A HCC has a greater possibility of containing slow-growing intrahepatic tumor clusters, and the removal of any rapidly growing tumors from among these should be undertaken by reduction surgery followed by subsequent multidisciplinary treatment for residual tumor cells, including appropriate immunotherapy. PMID- 8400671 TI - Procollagen type III N-peptide and type IV collagen 7S-domain in the sera of breast cancer patients. AB - We investigated the usefulness of serum levels of procollagen type III N-peptide (P III P) and the type IV collagen, 7S-domain, (IV-C) as markers of recurrent and metastatic breast cancer. Serum P III P and IV-C levels were found to be significantly higher in patients who showed postoperative recurrence, with 76.9% of P III P-positive cancer bearing patients and 68.0% of IV-C-positive cancer bearing patients having metastases in the bone and/or liver. Furthermore, 66.7% of all patients with metastases in the bone and/or liver were P III P-positive and 75.6% were IV-C-positive. Patients with liver metastases were either P III P- or IV-C-positive, and the levels were higher than those in patients with bone metastases. Thus, a positive correlation of serum P III P and IV-C levels was observed. These findings suggest that both serum P III P and IV-C levels are useful markers of recurrent and metastatic breast cancer, especially of disease in the bone and/or liver. PMID- 8400672 TI - A pharmacodynamic and pharmacokinetic study of fluoropyrimidines in a nude mouse system and in postoperative patients with gastric cancer. AB - To evaluate the effect of the oral fluoropyrimidines, tegafur and uracil (UFT) and 5-fluorouracil (5-FU), a pharmacodynamic analysis was conducted using a nude mouse system and patients. In the nude mouse system, UFT and 5-FU showed similar marginal effects against the human tumor xenograft Co-4, and the concentration of 5-FU in serum 1 h after the last administration being 0.04 micrograms/ml, which was assumed to be the minimum effective concentration. (MEC). Postoperative patients were subdivided into three groups, being: those who underwent subtotal gastrectomy and received UFT; those who underwent subtotal gastrectomy and received 5-FU; and those who underwent total gastrectomy and received UFT. In the UFT groups, the concentration of 5-FU in the portal and peripheral blood showed similar elimination in terms of the peak concentration (Cmax) and the area under the curve (AUC). In the 5-FU groups, the AUC and Cmax were significantly higher in portal blood than peripheral blood. The concentrations in the portal blood of the 5-FU group and in the portal and peripheral blood of the UFT group were significantly higher than the MEC (0.04 micrograms/ml). From these pharmacodynamic data, it was concluded that postoperative chemotherapy with oral fluoropyrimidines can achieve the MEC in portal and peripheral blood. PMID- 8400673 TI - A pathohistological and biochemical study of arteriosclerosis in the internal thoracic artery, a vessel commonly used as a graft in coronary artery bypass surgery. AB - Pathohistological and biochemical studies were conducted on the severity of arteriosclerosis in the internal thoracic artery (ITA), an artery commonly used for coronary artery bypass grafting (CABG). For the pathohistological examination, 26 bilateral ITAs and 13 left anterior descending coronary arteries (LADs) obtained in full length from 13 autopsy cases, none of which had died of arteriosclerotic heart disease, were used. The ratio of the thickness of the intima to that of the media (R) was used as the index for arteriosclerosis. ITAs and LADs were classified as grades I to IV according to the value of R. The R of the ITAs was approximately 1/10 that of the LADs (P < 0.01). Most ITAs showed a low arteriosclerotic grade, with no variation in arteriosclerosis along their length and a low R in all segments. No difference was found between right and left ITAs. Biochemical examination was conducted on 12 ITAs and 11 LADs, obtained from 12 different and unselected autopsy cases. The lipid content in the vascular wall was determined to evaluate the severity of arteriosclerosis, with the following results: Total cholesterol, 5.5 +/- 1.8 and 17.8 +/- 13.6 micrograms/mg wet weight (P < 0.05); triglyceride, 90.4 +/- 90.3 and 114.4 +/- 117.2 micrograms/mg wet weight (n.s.); and phospholipid, 7.4 +/- 3.9 and 11.2 +/- 3.9 micrograms/mg wet weight (P < 0.05), respectively, for the ITAs and LADs. These findings thus demonstrate that arteriosclerosis of the ITA in Japanese people is very mild, compared to that of the LAD in the same individuals. PMID- 8400674 TI - A comparison between ultrasonography and mammography, computed tomography and digital subtraction angiography for the detection of breast cancers. AB - Ultrasound (US) was compared with mammography (MMG), computed tomography (CT), and digital subtraction angiography (DSA) in its effectiveness to detect breast cancer masses and metastatic axillary nodes. Forty-seven breast cancer patients who all underwent MMG, US, CT, and DSA preoperatively in our institution between 1986 and 1990 were studied. US was able to detect tumors in all cases regardless of tumor size, whereas DSA detected T1-size tumors and MMG detected T2-size tumors in 40% and 64.7% of cases, respectively, being specifically inferior to US. It was found that MMG was least likely to detect papillotubular carcinoma, although microcalcification alone without a tumor mass on MMG improved detectability from 46.2% to 76.9%, according to the histological type. CT was found to be most sensitive to axillary node metastases (81.8%), followed by US (72.7%), but DSA was significantly unfavorable (42.9%). Thus, we concluded that US was superior to MMG, CT, and DSA for detecting breast cancer masses, but that CT was more advantageous than US, while DSA was of little value for evaluating axillary nodal status. PMID- 8400675 TI - Effect of indomethacin on postoperative protein metabolism after gastrectomy under total parenteral nutrition. AB - A randomized trial was undertaken to evaluate the effects of postoperative indomethacin (IDM) administration on protein metabolism in 20 patients who underwent an uncomplicated distal gastrectomy and were placed on post-operative total parenteral nutrition (TPN). Ten patients (the IDM group) received 50 mg of IDM every 8 h after operation up to postoperative day (POD) 4 while the other ten patients (the control group) received neither IDM nor any other non-steroidal anti-inflammatory drug, postoperatively. Though the requirement for postoperative plasma transfusion was significantly greater in the IDM group, the albumin level on POD 1 was significantly lower in this group than in the control group. The postoperative changes of C-reactive protein, retinol binding protein, and pre albumin between the two groups showed no difference. Moreover, the urinary 3 methylhistidine excretion, N-balance, and plasma aminogram on POD 4 also demonstrated no difference. We thus concluded that post-operative IDM administration after elective surgery has no additional anti-catabolic effect on the presence of TPN. PMID- 8400676 TI - A flow cytometric bromodeoxyuridine/DNA analysis for cytokinetics of the pancreatic cancer cell line Capan-2 after irradiation. AB - The cytokinetics of the human pancreatic adenocarcinoma cell line Capan-2 were analyzed by a bromodeoxyuridine (BrdUrd)/DNA analysis with flow cytometry (FCM). The reproducibility of data was shown to be superior to that achieved using a DNA analysis program with FCM. The perturbation of cytokinetics following irradiation was examined. Bivariate distribution was able to distinguish each cell phase cohort, even following as strong an irradiation as 10 Gy, a condition for which DNA analysis proved unusable. BrdUrd/DNA analysis showed the G2M phase population after irradiation with 2 Gy to be significantly greater (P < 0.005) than that of the untreated cells. This difference was not detectable by the DNA analysis program due to the large standard deviation (SD) of the data. BrdUrd/DNA analysis has a sufficient quantitative accuracy that strongly perturbed cytokinetics in patients treated with various therapeutic agents, for example, with combined therapies, can thus be analyzed by this approach. PMID- 8400677 TI - The effects of allopurinol and SOD on lipid peroxidation and energy metabolism in the liver after ischemia in an aerobic/anaerobic persufflation. AB - This study was aimed at examining the vulnerability of the liver to oxygen-free radicals upon reoxygenation after prolonged ischemia. Livers from male Wistar rats were first flushed with Ringer's and Euro-Collins solutions. After ischemic storage in Krebs-Henseleit solution at 37 degrees C for 60 min and in Euro Collins solution at 4 degrees C for another 60 min, they were then persufflated with either gaseous O2 or N2 for 30 min at 37 degrees C, and rinsed again with Ringer's solution. Enzyme concentrations and calcium ion activities were measured in the effluent rinsing solution after passage through the liver. Treatment with superoxide dismutase (SOD) or allopurinol resulted in a significant reduction of tissue injury, determined by the enzyme loss, calcium uptake, and lipid peroxidation upon persufflation with O2. Allopurinol also improved the tissue levels of ATP and the sum of adenine nucleotides after aerobic persufflation, whereas SOD did not. Notwithstanding, neither treatment had any effect on anoxic persufflation with N2. Thus, we conclude that the postischemic liver is susceptible to oxygen-induced free radical injury and that allopurinol and SOD promote specific antioxidative protection of the liver, with the exclusion of side effects related to substrates or perfusion modalities. PMID- 8400678 TI - An appraisal of segmental pancreatectomy for benign tumors of the pancreatic body: a report of two cases. AB - To preserve pancreatic and splenic function, segmental pancreatectomy with pancreatojejunostomy was performed on two patients with benign pancreatic tumors, and a 3-year follow-up study was conducted. The pancreatic endocrine functions assessed by 75 g oral glucose tolerance test were normal 3 years after surgery, and the exocrine functions returned to within the normal range 1 month after surgery according to a pancreatic function diagnostant (PFD) test. The platelet count increased transiently to 48.9 x 10(4)/mm3, and 70.5 x 10(4)/mm3, in the two patients, respectively, but returned to the preoperative value 1 month postoperatively. The operative times were 7 h 51 min and 5 h 3 min, which included the time taken for intraoperative ultrasonography and frozen section diagnosis, and the blood losses were 183 ml and 212 ml. The postoperative hospitalization period averaged 39 days and no complications developed in either patient. The method of performing segmental pancreatectomy, initially reported by Letton and Wilson for cases of pancreatic trauma, was evaluated and successfully applied to benign pancreatic tumors. PMID- 8400679 TI - Obstructive jaundice associated with extrahepatic portal vein obstruction: report of two cases. AB - We herein report two cases of obstructive jaundice with markedly dilated collateral veins either in or around the bile duct in the setting of extrahepatic portal vein obstruction (EHPO). In the first case, a proximal splenorenal shunt provided relief of biliary stenosis as well as eradication of esophageal varices due to a decompression of portal hypertension. This evidence proved that the markedly extended collateral veins in the hepatoduodenal ligament caused biliary stenosis by compressing the bile duct. In the second case, obstructive jaundice was probably caused by cholangitis and was relieved with biliary drainage. Portal decompressive surgery was not indicated because of the slight degree of esophageal varices. The relationship between cholangitis and EHPO in these patients calls for further investigation. In cases with EHPO manifesting obstructive jaundice associated with risky esophageal varices, portal decompressive surgery is recommended as the procedure of choice. PMID- 8400680 TI - Adult neuroblastoma in pregnancy: report of a case. AB - We report herein an extremely rare case of retroperitoneal neuroblastoma occurring in a mother during her third trimester of pregnancy. A live male infant was delivered by cesarean section at 36 weeks gestation and 23 days later, the mother's tumor was curatively resected. Six months have elapsed since her operation and there has been no evidence of tumor recurrence. Adult neuroblastoma is an uncommon malignancy and, to our knowledge, there has been no case of this tumor occurring in pregnancy ever described in the English literature. The diagnosis, treatment, and follow-up of this neoplasm and its association with pregnancy are discussed in this report. PMID- 8400681 TI - Gas-forming liver abscess after transcatheter arterial embolization for hepatocellular carcinoma: report of a case. AB - A case of a gas-forming liver abscess developing after transcatheter arterial embolization for recurrent hepatocellular carcinoma (HCC) in a 65-year-old man is presented herein. He was admitted to hospital with fever and jaundice, following which ultrasonography (US) and computed tomography revealed a gas-containing abscess in the posterior segment of the hepatic lobe with multiple HCC. Percutaneous transhepatic drainage was performed using US. Antibiotics which were sensitive to the Escherichia coli bacteria detected in the abscess were administered both intravenously and through the drainage tube into the abscess. Four months later, the abscess had diminished and the patient was discharged after receiving percutaneous ultrasonographically guided ethanol injection therapy for the recurrent HCC. PMID- 8400682 TI - Branch retinal artery occlusion following thyroidectomy for papillary carcinoma of the thyroid: report of a case. AB - A rare case of branch retinal artery occlusion (BRAO) following a subtotal thyroidectomy for thyroid cancer in a 58-year-old woman is reported herein. Five days after her thyroidectomy, the patient complained of having had a reduction in visual acuity and visual field loss of the superior nasal side in her right eye since the operation. BRAO was diagnosed following the discovery by funduscopy of inferotemporal branch artery occlusion with retinal edema, hemorrhage, and periarterial sheathing in the right eye. Despite immediately puncturing the anterior camera and massaging the eyeball while administering intravenous anticoagulant therapy, the visual field disturbance remained unchanged. The most common causative factor of postoperative sudden BRAO is reported to be emboli. However, in our case, the most likely cause was the stretching and pressure exerted on the carotid artery with consequent atheromatous plaque formation at the time of thyroidectomy. PMID- 8400683 TI - Brief introduction to human DNA fingerprinting. PMID- 8400684 TI - Rapid analysis of PCR components and products by acidic non-gel capillary electrophoresis. AB - A capillary electrophoresis system has been developed which has the ability to rapidly analyse DNA restriction fragments, PCR products, oligonucleotides and complex deoxyribonucleoside tri-, di- and mono-phosphate mixtures in a single separating medium. Separations are performed in an internally coated capillary containing a solution of linear polymers. The separation of all DNA species is achieved through the novel use of acidic rather than alkaline pH. This has the added advantage of preserving the internal capillary coating. The use of the technique is described for rapid, high resolution separation of pBR322 and phi X174 DNA restriction digests, quality control of dNTP and oligonucleotide primer PCR components, detection of a PCR amplified region of lambda-phage DNA and detection of a PCR amplified human hypervariable region of forensic interest. The technique is termed "acidic non-gel capillary electrophoresis". PMID- 8400685 TI - Minisatellite variant repeat mapping: application to DNA typing and mutation analysis. AB - Most DNA typing systems assay allele length variation at tandemly repeated loci such as minisatellites and microsatellites. Allele length measurements are approximate, which impedes the use of such loci in forensic analysis and in studies of allelic variability at hypervariable loci. We now review progress in the development of alternative DNA typing systems based on allelic variation in the interspersion patterns of variant repeat units along minisatellite alleles. Minisatellite variant repeat mapping by PCR (MVR-PCR) not only provides a powerful new digital approach to DNA typing, but also for the first time allows investigation of the true level of allelic variability at minisatellite loci and of the mutational mechanisms that generate ultravariability. PMID- 8400686 TI - Microsatellites and disease: a new paradigm. PMID- 8400687 TI - A unified approach to study hypervariable polymorphisms: statistical considerations of determining relatedness and population distances. AB - Relatedness between individuals as well as evolutionary relationships between populations can be studied by comparing genotypic similarities between individuals. When hypervariable loci are used to describe genotypes, it is shown that both of these problems can be approached with a unified theory based on allele sharing between individuals. The distributions of the number of shared alleles between individuals indicate their kin relationships. Extending this, we obtain statistics for genetic distances between populations based on average number of alleles shared between individuals within and between two different populations. Traditional statistical inferential procedure can be used to establish specific kinship relationships between individuals. We derive estimates of the number of hypervariable loci needed for a specified reliability of such an inference. Evolutionary dynamics of genetic distance statistics based on allele sharing is also studied. It shows that such measures of genetic distances remain linear with the time of divergence for a period comparable to that of the gene frequency-based measures of genetic distances. Statistical properties of measures based on allele sharing establish that for using such summary statistics it is not necessary to know the full characteristics of all loci used. It is enough to know the degree of heterozygosity per locus and the number of loci. Therefore, in principle, this approach can also be used for DNA fingerprinting data in the studies of relatedness between individuals as well as between populations. The possible compromising features of multilocus DNA fingerprinting data are also discussed. PMID- 8400688 TI - The forensic significance of various reference population databases for estimating the rarity of variable number of tandem repeat (VNTR) loci profiles. AB - The likelihood of occurrence of 1,964-HaeIII-generated target DNA profiles was estimated using fixed bin VNTR frequencies from various Caucasian, Black, and Hispanic databases and the product rule. The data in this study demonstrate that for forensic purposes there are smaller differences in statistical estimates of DNA profile frequencies among subgroup databases than among estimates across major population databases. This observation does not support the premise asserted by the NCR Report (1992) that the differences among subgroups within a race would be greater than between races (at least for forensic purposes). Therefore, the data do not support the need for alternative procedures, such as the ceiling principle approach (NRC Reports, 1992), for deriving statistical estimates of DNA profile frequencies. Comparisons across major population groups provide reasonable, reliable, and meaningful estimates of DNA profile frequencies without forensically significant consequences. PMID- 8400689 TI - Notes on the definition and nomenclature of tandemly repetitive DNA sequences. AB - Tandemly repetitive DNA is a major component of all eukaryotic genomes. This fact has been known for almost 30 years and research on this class of DNA is still being done. Its biology and evolution are therefore now becoming fairly well understood. DNA-fingerprint techniques rely very much on this knowledge. However, the large amount of research on these sequences has inevitably led to a large number of different concepts and theories about their nature. This has also resulted in some confusion as to the nomenclature. The following notes are intended to resolve this confusion somewhat and to give some definitions for the major classes of tandemly repetitive DNA. PMID- 8400690 TI - Genetic variation among the Mapuche Indians from the Patagonian region of Argentina: mitochondrial DNA sequence variation and allele frequencies of several nuclear genes. AB - DNA samples from 60 Mapuche Indians, representing 39 maternal lineages, were genetically characterized for (1) nucleotide sequences of the mtDNA control region; (2) presence or absence of a nine base duplication in mtDNA region V; (3) HLA loci DRB1 and DQA1; (4) variation at three nuclear genes with short tandem repeats; and (5) variation at the polymorphic marker D2S44. The genetic profile of the Mapuche population was compared to other Amerinds and to worldwide populations. Two highly polymorphic portions of the mtDNA control region, comprising 650 nucleotides, were amplified by the polymerase chain reaction (PCR) and directly sequenced. The 39 maternal lineages were defined by two or three generation families identified by the Mapuches. These 39 lineages included 19 different mtDNA sequences that could be grouped into four classes. The same classes of sequences appear in other Amerinds from North, Central, and South American populations separated by thousands of miles, suggesting that the origin of the mtDNA patterns predates the migration to the Americas. The mtDNA sequence similarity between Amerind populations suggests that the migration throughout the Americas occurred rapidly relative to the mtDNA mutation rate. HLA DRB1 alleles 1602 and 1402 were frequent among the Mapuches. These alleles also occur at high frequency among other Amerinds in North and South America, but not among Spanish, Chinese or African-American populations. The high frequency of these alleles throughout the Americas, and their specificity to the Americas, supports the hypothesis that Mapuches and other Amerind groups are closely related.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400691 TI - Microsatellite and HLA class II oligonucleotide typing in a population of Yanomami Indians. AB - We have used three different microsatellites (on chromosome 12 and Y) together with HLA class II oligonucleotide typing (DQA and DQB) to analyze families of Yanomami indians settling in villages in Southern Venezuela. There exist complex networks of biological relationship between villages as a result of wife exchange, village fissioning and changing patterns of alliances associated with inter-village warfare. Social status in this society is largely determined by the kinship system. Polygyny is common, especially among headmen, with additional wives, frequently being chosen among the sisters of the first wife. Our preliminary results mainly obtained from inhabitants of the village HAP show the expected allele distribution in populations with a high degree of consanguinity: (i) deficiency of observed heterozygotes at the autosomal loci and (ii) almost all men carry the same Y chromosomal allele. Nevertheless in the Yanomami village two thirds of the described autosomal microsatellite alleles were identified. Several paternities were clarified. PMID- 8400692 TI - Iserables: a Bedouin village in Switzerland? AB - Iserables is an alpine village--about 1000 inhabitants--which remained isolated till these recent years because of its particular geographical situation. The Iserables inhabitants call themselves "Bedjuis" (Bedouin in local dialect) and regard themselves as descendants of the Sarrazins who invaded the Alps during the VIII-X centuries. Our goal, in studying several DNA-VNTR polymorphims, in addition to some blood groups, within the Iserables community, was to see if there was any evidence supporting this popular belief. As a preliminary phase of this project, the allelic frequencies for six VNTR loci analysed for 102 individuals of the village (all descendants of nine original families) are presented. The results are compared with those reported for Swiss and white populations. PMID- 8400693 TI - Paternity testing with the F10 multilocus DNA fingerprinting probe. AB - Empirical analysis of 200 paternity cases by multilocus DNA fingerprinting with the F10 probe showed that it was capable of distinguishing fathers from non fathers in every case. The average exclusion probability was 0.99998. A very effective discrimination parameter was the proportion of non-maternal (test) bands which cannot be detected in the alleged father (unassignable bands) among all test bands. Values below 0.2 were seen in true fathers while in all cases of non-fathers the values were above 0.35. Minisatellite mutations occurred at a rate of 0.004 per band per child. The distribution of band-sharing among first degree relatives and unrelated individuals showed only a small overlap. Thus, band-sharing of the F10 fingerprints should provide a useful statistic for testing genetic relationships in deficiency cases. PMID- 8400694 TI - The formal analysis of multilocus DNA fingerprints. AB - A description is given of a novel method for the formal analysis of multilocus DNA fingerprints, the so-called 'genetic factor model'. Using this model, multilocus DNA fingerprints can be shown to be a robust means for both paternity testing and pedigree reconstruction. PMID- 8400695 TI - Oligonucleotide DNA fingerprinting: results of a multi-center study on reliability and validity. AB - We report the results of an empirical study of 256 paternity cases referred to 7 different German laboratories for DNA fingerprinting with oligonucleotide probe (CAC)5/(GTG)5. All parameters characteristic of such multilocus DNA fingerprints were found to differ significantly between the contributing centres. Despite these differences, clear-cut decisions between paternity and non-paternity could be made in all but one case. Furthermore, we found no systematic deviation of the gel-phenotype distribution among trios from random expectation as derived from commonly adopted analytical models. Thus, we conclude that oligonucleotide DNA fingerprinting is a robust and reliable means for the resolution of paternity cases. PMID- 8400696 TI - Testing deficiency paternity cases with a Y-linked tetranucleotide repeat polymorphism. AB - Because a son's genotype at a Y-linked locus uniquely specifies his father's genotype at that locus irrespective of the maternal contribution, Y-linked polymorphisms show increased exclusion power over autosomal polymorphisms in paternity cases involving a male child. This advantage is even more obvious when the alleged father is deceased or otherwise unavailable for testing. In this situation, any individual connected by patrilineage to the alleged father may be tested in his place. The usefulness of the Y-linked tetranucleotide repeat locus Y-27H39 in deficiency cases was evaluated in a set of 41 families that had a deceased alleged father and that had been tested at GENE/MG with multilocal and unilocal DNA fingerprinting probes. In sixteen of these cases the proband (child tested) was male and there were male relatives. In the thirteen cases in which paternity was included, Y-27H39 would contribute significantly to the paternity index. In one of three cases in which there was exclusion by fingerprinting probes there was also exclusion by Y-27H39. Thus, Y-27H39 is useful in deficiency paternity cases and will be specially valuable in situations where autosomal polymorphisms have limited power, such as when there is only one male living relative of the deceased father. However, our experience is that Y-linked probes can only be applied in approximately 40% of cases. PMID- 8400697 TI - Short tandem repeat loci: application to forensic and human remains identification. AB - The short tandem repeat (STR) locus ACTBP2 (common name SE33) was analyzed for its potential use in forensic and human remains identification. PCR amplification conditions were determined, and an allele-specific ladder was generated so that discrete alleles could be scored. The allele frequency distributions were determined for both Caucasian and Black populations. The frequency data meets Hardy-Weinberg expectations, and the allele distributions were similar from one racial group to another and between ethnic groups. SE33 analysis was subsequently used to confirm the identification of human remains for the Office of the Armed Forces Medical Examiners. PMID- 8400698 TI - Forensic DNA typing by the solid-phase minisequencing method. AB - We describe a method for DNA-typing, in which a panel of biallelic markers are detected by the solid-phase minisequencing method (Syvanen et al., 1990). This method identifies single nucleotide variations in DNA fragments amplified by the PCR. Determination of the panel of 12 markers selected in this study proved to be an efficient and reliable method for forensic identification of individuals. We also introduce a novel approach for rapid determination of allele frequencies by quantitative analysis of pooled DNA samples. PMID- 8400699 TI - The use of polymorphic Alu insertions in human DNA fingerprinting. AB - We have characterized several Human Specific (HS) Alu insertions as either dimorphic (TPA25, PV92, APO), slightly dimorphic (C2N4 and C4N4) or monomorphic (C3N1, C4N6, C4N2, C4N5, C4N8) based on studies of Caucasian, Asian, American Black and African Black populations. Our approach is based upon: 1) PCR amplification using primers complementary to the unique DNA sequences that flank the site of insertion of the different Alu elements studied; 2) gel electrophoresis and scoring according to the presence or absence of an Alu insertion in one or both homologous chromosomes; 3) allele frequencies determined by gene counting and compared to Hardy-Weinberg expectations. Our DNA fingerprinting procedure using PCR amplification of diallelic polymorphic (dimorphic) Human Specific Alu insertions, may be used as a tool for genetic mapping, to characterize populations, study human migrational patterns, and track the inheritance of human genetic disorders. PMID- 8400700 TI - On the essence of "meaningless" simple repetitive DNA in eukaryote genomes. AB - Various kinds of simple tandemly repetitive DNA sequences are abundantly interspersed in the genomes of practically all eukaryotic species studied. The comparatively elevated mutation rates of simple repeat blocks result in highly polymorphic and therefore extremely informative investigation systems for studies on forensic, ecological and genetic relationship questions. Recently the techniques for analyzing simple repeats have achieved great effectivity and simplicity. Beyond their utility as tools for differentiation and individualization, certain of these repeated elements harbor quite unexpected qualities which may be discussed in the context of their biological meaning. i) A specific subset of simple (cac)n or (gtg)n repeats is expressed in mature mRNA and total cellular RNA. ii) Despite the apparently high mutation rate certain (gt)n or mixed (gt)n/(ga)m stretches of intronic simple repeats are preserved in immunologically relevant genes for at least 70 x 10(6) years and they bind nuclear protein molecules with high affinities. Consequently in addition to their tool character, the biological aspects of simple repeated DNA should be taken into consideration. PMID- 8400701 TI - DNA- and PCR-fingerprinting in fungi. AB - DNA-fingerprinting has been successfully used to detect hypervariable, repetitive DNA sequences (minisatellites and microsatellites) in fungi. Combined with methods used to identify random amplified polymorphic DNA (RAPD), conventional DNA-fingerprinting hybridization probes can also be used as single primers to detect DNA polymorphisms among fungal species and strains. The oligonucleotides (CA)8, (CT)8, (CAC)5, (GTG)5, (GACA)4 and (GATA)4, as well as the phage M13 and its core sequence, have been used as specific probes in hybridization experiments and as primers for PCR analysis. Both methods have enabled the differentiation of all the fungal species and strains that were examined, including species of Penicillium, Trichoderma, Leptosphaeria, Saccharomyces, Candida and Cryptococcus. These methods have been used 1) to clarify the taxonomic relationships among relevant species of the Trichoderma aggregate, 2) to discriminate between aggressive and non-aggressive isolates of the rape seed phytopathogen, Leptosphaeria maculans, and 3) to identify strains of the pathogenic yeasts, Cryptococcus neoformans and Candida albicans. PCR-fingerprinting allowed serotypes of C. neoformans to be distinguished. The application of DNA- and PCR fingerprinting to fungal DNA should aid in clarification of their taxonomy and improved diagnosis of mycotic disease. PMID- 8400702 TI - DNA fingerprinting reveals relationships between strains of Trypanosoma rangeli and Trypanosoma cruzi. AB - Very little is known about the structure and sequence of the genomic DNA and kDNA of T. rangeli and no highly polymorphic markers are known. In this paper, we show that the Jeffreys' multilocal probe 33.15 produces characteristic DNA fingerprints with these trypanosomes. The multiband patterns can be used to differentiate T. cruzi from T. rangeli and for recognizing relationships between strains of the latter from widely different geographic areas and different hosts. The topology of a UPGMA phenetic tree constructed from band-sharing data suggests the existence of two groups of T. rangeli: one encompassing parasites from Central America and the northern part of South America and another with the parasites from southern Brazil. This splitting was confirmed by the use of both nuclear and kinetoplast unique sequence probes. Among strains of T. rangeli, band sharing was generally negatively correlated with geographical distance. This work confirms the usefulness of DNA fingerprints as a potent technique for the analysis of relationships in trypanosomatid populations. PMID- 8400703 TI - The use of RAPDs for the analysis of parasites. AB - There is a lack of sequence information concerning polymorphic loci in parasite genomes. Thus, the use of arbitrary PCR primers under low temperature annealing conditions to generate random amplified polymorphic DNAs (RAPDs) represents an important approach to the study of the structure of parasite populations, their genetic variation as well as improved diagnosis of the diseases they cause. Following the examination of all variables and their effect on the reproducibility of the reaction, we have established a protocol for the analysis of RAPDs that involves amplification at two separate DNA concentrations followed by polyacrylamide gel electrophoresis and silver staining. We find the technique to be sensitive, reproducible, simple and relatively cheap. It has already provided insight into the genetic variation in populations of schistosomes and trypanosomes and is being used to study various other endemic infections. We also use specific primers under low stringency conditions in situations where the objective of the amplification is the detection of a particular sequence and where normal high stringency conditions give a positive/negative answer such as sex determination or diagnosis of blood born infections. Under low stringency conditions, specific amplification products persist but products of low stringency priming are also apparent and serve as a perfect internal control for negative samples. PMID- 8400704 TI - The use of RAPDs for the study of the genetic diversity of Schistosoma mansoni and Trypanosoma cruzi. AB - Arbitrary primers have been used for the production of complex, PCR generated DNA profiles in order to undertake a preliminary random amplified polymorphic DNA (RAPD) analysis of strains (and related species) of two parasitic organisms that are responsible for important diseases endemic in Brazil: Schistosoma mansoni that causes schistosomiasis, and Trypanosoma cruzi that causes Chagas' disease. A relatively low level of polymorphism was found in S. mansoni when strains isolated from different regions of Brazil were compared, with less than 10% of bands exhibiting polymorphism. Comparison of different schistosome species, on the other hand, showed them to be distantly related with very few bands shared by even the more closely related species. Trypanosome strains were found to be much more variable. When strains were compared between zymodemes (groups of parasite strains with the same isoenzyme profiles), a maximum of 7% of bands were found to be common whereas among strains in the same zymodeme a clear characteristic pattern was observed. In the zymodeme most thoroughly studied, it was found that 59% of bands were shared. Band sharing analysis showed that the relationships of strains within a zymodeme correlate with their geographical origin and that the relationship between zymodemes correlates closely with that previously determined by isoenzyme analysis. These preliminary data indicate the ready applicability of RAPD analysis to the study of parasites where largely unexplored genetic variations may have an important bearing on the complexity and diversity of diseases. PMID- 8400705 TI - Quantitative traits in chicken associated with DNA fingerprint bands. AB - In an unselected control line of a selection experiment in chicken the following traits were evaluated: number of eggs, average egg weight, egg mass, age at first egg, body weight at 20 weeks, body weight at 40 weeks, feed consumption and feed efficiency (feed consumption/egg mass). Data and blood samples of 143 females of the same age were available for analysis. The animals were ranked for each trait according to the phenotypic performance adjusted for hatch and laying house effect rather than the breeding value. DNA mixes of the top 5 and the bottom 5 hens were compared to each other by DNA fingerprinting. The most striking differences could be observed with the probe pV47 in the mixes for body weight at 20 weeks and the feed efficiency. In the following the study focused on feed efficiency. Five groups of 10 hens each, around quantiles 0%, 25%, 50%, 75% and 100%, were analyzed with respect to the occurrence of 7 particular DNA fingerprint bands. The association of the groups' mean feed efficiency with the groups' band frequencies was assessed by linear regression. For 2 bands significant regression coefficients (P < 0.005) were found. PMID- 8400706 TI - Paternity testing of endangered species of birds by DNA fingerprinting with non radioactive labelled oligonucleotide probes. AB - In the last years, DNA fingerprinting became the most powerful tool for identification and paternity testing in man. The success of this method encouraged the German Federal Ministry of Environment, Natural Protection and Reactor Safety to apply DNA fingerprinting in the field of protection of endangered species of birds, such as birds of prey or parrots. In the last three years, we received more than 400 blood and tissue samples of 23 species of birds of prey or parrots, most of them obtained by confiscation, to establish paternity and legal breeding success. We used digoxigenated oligonucleotide probes, mainly (GGAT)4 and (GACA)4 for hybridization. In most cases of confiscated families of birds, paternity testing showed exclusions of nestlings. PMID- 8400707 TI - DNA fingerprinting of trait-selected mouse lines and linkage analysis in reference families. AB - The first aim of the study was the molecular genetic characterization of long term trait-selected (growth, adaptability and fertility) lines of mice using multilocus DNA fingerprinting with the simple tandem repetitive oligonucleotide probes (GAA)6 and (GACA)4. Secondly polymorphic markers were screened for association with growth performance based on DNA fingerprint analysis in reference families. Pooled DNA samples of ten unrelated mice of trait-selected mouse lines (over 40 generations) and the unselected control lines were analyzed. Resulting differences in band patterns were reanalyzed comparing individual fingerprints of the animals included in the pooled DNA samples of the different lines. Between the mouse lines about 30% of analyzed fingerprint bands were polymorphic. Individuals of long-term selected mouse lines show only a few individual-specific bands. Most polymorphic bands observed in DNA mixes appear in all animals included in the DNA mix of the corresponding line. Line-specific DNA fingerprint bands were analyzed for their inheritance and linkage with growth performance in reference families using animals with extreme growth performance of the first backcross after crossing of growth-selected with unselected mice. Scanning the distribution of line-specific bands in the reference panel few bands were identified which are associated with growth performance. They appear as useful markers for growth selection. Nevertheless most line-specific bands result from genetic drift rather than from selection. PMID- 8400708 TI - Dog genetic polymorphism revealed by synthetic tandem repeats. AB - We are studying the genetic polymorphism associated with Variable Number of Tandem Repeat (VNTR) loci in 13 breeds of dogs, namely: Alaskan Malamute, Barzoi, Beagle, Belgian Shepherd, Fox Terrier, Griffon, Labrador, Irish Setter, Spaniel, Dachshund, Irish Terrier, Shar Pei and Poodle. Our approach is based upon synthetic tandem repeats (STRs). Using a panel of these arbitrary unit polymers to detect minisatellites, we are attempting to develop paternity testing systems on pure bred dog pedigrees. We are evaluating the potential importance of STRs as a tool for the isolation of minisatellites in dogs, as well as for the characterization of dog genetic markers. PMID- 8400709 TI - Characterization of canine microsatellites. AB - Canine DNA was cloned in M13 and screened for the presence of (dC-dA)n.(dG-dT)n repeats. Oligonucleotide primers were synthesised to the microsatellite flanking sequences and used in the polymerase chain reaction to amplify those loci from genomic DNA. The polymorphism of each microsatellite was estimated in a set of unrelated dogs. PMID- 8400710 TI - Application of human minisatellite probes to the development of informative DNA fingerprints and the isolation of locus-specific markers in animals. AB - In this study, the alpha-globin 3' HVR (Jarman et al., 1986), the RNA transcripts of 33.15 and 33.6 (Carter et al., 1989), and the human locus-specific minisatellites MS1, MS8, MS31, MS32 (Wong et al., 1987), MS51, MS228A (Armour et al., 1989) and g3 (Wong et al., 1986) were applied to domestic pigs, common marmoset monkeys (Callithrix jacchus jacchus) and Waldrapp ibises (Geronticus eremita) and evaluated for their suitability firstly for isolating polymorphic VNTR markers from genomic libraries (pigs), and secondly for producing informative DNA fingerprints (pigs, marmosets and Waldrapp ibises). PMID- 8400711 TI - The 'individualization' of large North American mammals. AB - The enforcement of wildlife laws and the captive breeding of threatened/endangered species requires the ability to identify individual animals. DNA profiles of a variety of large North American mammals, birds, and fish were generated using ten different oligonucleotide probes. The probes tested were four multilocus probes [33.6, 33.15, JE46, and (TGTC)5] and six 'human unilocus' probes [MS1 (D1S7), CMM101 (D14S13), YNH24 (D2S44), EFD52 (D17S26), TBQ7 (D10S28), and MS43 (D12S11). Each of the probes was chemically synthesized, and labeled by the attachment of alkaline phosphatase; after hybridization, the probes were detected by chemiluminescence catalyzed by the enzyme. Initial screening against zoo blots including samples of bear, wolf, large cat, wild sheep, deer, birds, marine mammals, and fish indicated that three multilocus probes [33.15, 33.6, (TGTC)5] gave informative patterns containing 15-40 bands for most or all of the animals tested, as did two of the 'human unilocus' probes (MS1 and CMM101). The other five probes appeared informative only in some species (for example, YNH24 against canids). Subsequent screenings of populations within species were used to determine genetic diversity by analysis of observed bandsharing (S). Large heterologous populations, such as white-tailed deer, exhibited highly diverse band patterns (S < or = 0.2). Geographically isolated and/or genetically constricted animals, such as endangered Mexican wolves, Tule elk, and Columbian white-tailed deer, exhibited much higher frequencies of bandsharing (0.6 < or = S < or = 0.95).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400712 TI - Bovine microsatellites: racial differences and association with SINE-elements. AB - Patterns of polymorphism at eight microsatellite loci in three cattle races are described: two large commercial breeds and one endangered landrace. Significant interracial allele frequency differences were found at six loci. The mean heterozygosity was slightly higher in the landrace (H = 0.75) than in the others (H = 0.69). The difference is smaller than that found by DNA-fingerprinting. Intrapopulation distributions of microsatellite allele size (dinucleotide repeat number) were generally bimodal, with certain intermediate repeat types lacking. Sequencing of individual alleles revealed some hidden heterogeneity: an allele defined by PCR-product size actually corresponded to different sequence motifs in different races. Two of the microsatellites occurred as tails of SINE-elements. In contrast to some earlier reports, the position of one of the amplification primers within a high-copy-number SINE-element did not disturb microsatellite amplification; even multiplex PCR was possible. PMID- 8400713 TI - Oligonucleotide fingerprinting of free-ranging and captive rhesus macaques from Cayo Santiago: paternity assignment and comparison of heterozygosity. AB - Multilocus DNA fingerprinting with oligonucleotide probes (GTG)5, (GATA)4, and (CA)8 was applied in order to determine paternity in one birth cohort (15 infants) of social group (S) from the free-ranging colony of rhesus macaques (Macaca mulatta) on Cayo Santiago. While sires could be identified in 11 cases, all males tested (N = 19) could be excluded from paternity for the remaining four infants. Data revealed marked discrepancies between actual paternity and paternity as inferred from the observation of copulation behavior. Thus, a dominant social rank does not appear to be strongly associated with reproductive success. Furthermore, alternative reproductive strategies were found to yield comparable net benefits in reproduction. A second group of animals (M) was translocated from Cayo Santiago to the Sabana Seca Field Station in 1984. They have continuously resided together in a large outdoor enclosure since then. Here paternity assessment was seriously impeded by a reduced number of discriminating bands, i.e. offspring bands which were unequivocally derived from the sires. This was initially held to be indicative of a smaller degree of heterozygosity in Group M, and was attributed to inbreeding due to a lack of male immigration or extra-group fertilizations. However, a comparison of the DNA fingerprint patterns obtained in Group S and Group M lends only partial support to this idea. PMID- 8400714 TI - Detection, cloning, and distribution of minisatellites in some mammalian genomes. AB - The chromosomal distribution of minisatellites (cloned and/or detected using natural or synthetic tandem repeats) is strikingly different in man and mouse. In man, the vast majority is clustered in the terminal band of a subset of chromosome arms. Interestingly, the class of shorter tandem repeats called microsatellites is widespread along the chromosomes, suggesting that minisatellites can be created or maintained only in certain regions. In order to gain a better knowledge of these areas, we have studied a sub-telomeric cosmid from the pseudoautosomal region. Sixty kilobases of human genomic DNA starting approximately 20 kilobases from the human sex chromosomes telomere have previously been independently isolated in two cosmid clones (locus DXYS14) (Cooke et al., 1985); Rouyer et al., 1986). We have studied in more detail one of the two cosmids from this locus and found that it contains four different minisatellite structures representing 20 kilobases of the cosmid. These structures are unrelated to each other or to the minisatellite family described by Jeffreys et al. (1985). They display different degrees of polymorphism correlated with varying amounts of inner homogeneity. Combined with the previous description of an additional minisatellite (Cooke et al., 1985; Inglehearn and Cooke, 1990) in the contiguous cosmid, our observation shows that these structures may represent an important proportion of the DNA in sub-telomeric regions. PMID- 8400715 TI - Human VNTR mutation and sex. AB - Seven hypervariable VNTR loci have been studied in about 1200 parent/child pairs about equally divided between the sexes. Mutations were observed with all seven probes, the total number being 71. Fourty-four of these involved increased fragment length. Gains in fragment length were on average larger than losses. These findings indicate that mutation might be a basis for evolutionary expansion of VNTR fragment length. For five probes YNH24 (D2S44), MS31 (D7S21), g3 (D7S22), MS43A (D12S11), and CMM101 (D14S13), mutation rates were relatively low (less than 1%) with no obvious sex difference. MS1 (D1S7) mutation frequencies were substantially higher, with a tendency towards a higher paternal than maternal mutation rate (5.4% and 2.0%, respectively). The probe B6.7 (provisionally assigned to chromosome 20) exhibits about five times higher paternal than maternal mutation rates. The mutation rate of 7.6% in paternal chromosomes is among the highest reported in any VNTR locus. These findings could indicate that while low-mutant VNTRs might reflect meiotic crossover, mutation events in high mutant loci could more often be caused by other mechanisms during cell division. PMID- 8400716 TI - Variation of minisatellites in chemically induced mutagenesis and in gene amplification. AB - A mutation assay in cultured mammalian cells was developed based on direct analysis of minisatellite DNA. Chinese hamster cells (V79) were mutagenized with nitrosoguanidine and independent colonies were isolated and expanded. DNA fingerprints were then obtained after digestion with HinfI or HaeIII and hybridization with 33.15 and 33.6 probes (Jeffreys et al., 1985). 12 colonies from untreated cells were also analyzed. Digestion with HaeIII and hybridization with 33.15 probe detected the highest frequency of induced variants. The results suggest that minisatellite sequences are hypermutable sites that can be used to monitor the mutagenic effect of chemical agents. We have also analyzed the DNA fingerprints of 17 independent Chinese hamster (CHO) cell lines carrying amplification of the CAD gene. The DNA fingerprint analysis showed a variation in minisatellite regions in 3 lines while no variation was observed in independent colonies from the CHO parental cell line. The results suggest that these sequences may be hot spots for recombination during gene amplification. PMID- 8400717 TI - Iterons of stringently controlled plasmids and DNA fingerprinting. AB - DNA probes which detect polymorphic, repetitive sequences in a variety of genomes have been developed using different approaches. Naturally occurring plasmids, with repeated units termed iterons near or within their origins (ori) or replication, could be of interest for the development of probes, possibly even revealing novel minisatellite families in mammals involved in replicational processes. We used the plasmids P1, pSC101 and RSF1010 or their PCR amplified ori regions as probes in Southern blot hybridisations with mammalian DNA. At low stringency they generated reproducible fingerprint-like patterns. A bovine genomic library was screened at the same stringency with the PCR-amplified ori region of P1 containing the five times repeated core sequence 5'-ATGTGTGNTGNNGGG 3' to generate a probe for cattle DNA with higher specificity. PMID- 8400718 TI - Towards covering immunological genes with highly informative markers: a trans species approach. AB - To establish a highly informative screening system for immunologically relevant genes ("immunoprinting") we co-amplified via polymerase chain reaction (PCR) polymorphic exons plus adjacent intronic simple repetitive dinucleotide stretches in the T-cell receptor (Tcr) Vb6 and Major Histocompatibility Complex (MHC)-DRB loci in man and several ungulate species. In both gene families the basic structure of the simple repeat was found to be preserved for more than 70 x 10(6) years in all investigated species. The simple repeats exhibit extensive length variability. Distinct exon sequences are correlated with a defined repeat length and substructure. In addition, PCR and the oligonucleotides for typing were applicable to a broad range of species from different mammalian orders. Multiplex PCR of different members of the Tcr Vb6 family and MHC-DRB resulted in a complex pattern similar to an oligolocus fingerprint. Hence immunoprinting can be employed for searching for associations of immunologically relevant genes with diseases even across species barriers. PMID- 8400719 TI - Autism and related behaviours. AB - Childhood autism is conceptualized as a behavioural syndrome with several different aetiologies. Its prevalence is estimated at about one in every thousand children born. The male:female ratio is considerably higher than in the general population, but possibly not as high as 3-4:1 (as has been suggested for almost 50 years in the literature). A number of specific medical conditions are associated with autism and a comprehensive medical work-up is required in all cases with pervasive autistic symptomatology. Genetic factors contribute to the development of autism in some cases. Recent neurobiological and psychological studies contribute to a concept of the autistic syndrome as but one of several different syndromes characterized by impaired empathy skills. PMID- 8400720 TI - Overestimation of mentally retarded persons' IQs using the PPVT: a re-analysis and some implications for future research. AB - Numerous validity studies have shown that the PPVT consistently overestimates mentally retarded persons' IQs. One possible interpretation is that this phenomenon is an outcome of the dissociation between their cognitive level and their experience. Indeed, compared to intellectually average subjects of the same mental age, they have had more learning opportunities, simply because they have lived longer. In order to validate this hypothesis, the French version of the PPVT, Raven's Coloured Progressive Matrices (RCPM), and the 1966 version of the Binet-Simon were administered to 90 subjects of various chronological ages matched on mental age (30 nonretarded 5 year olds and two groups of 30 retarded subjects aged 10 and 16 years, respectively). The results indicate that CA exerts a strong effect on vocabulary, but not on RCPM performance. The research implications of this finding are discussed. PMID- 8400721 TI - Normalized growth velocity in children with Down's syndrome during growth hormone therapy. AB - Between 6 months and 3 years of age, growth velocity in children with Down's syndrome (DS) is markedly reduced in comparison to that of healthy children. However, after 3 years of age, it is almost normal. Thus, growth retardation becomes pronounced during the period when growth hormone (GH) starts to regulate growth. The present authors report the long-term effects of GH-therapy in 16 children with DS, who are being treated for 3 years from the age of 6-9 months. The treatment, Genotropin, 0.1 U kg-1 BW day-1, was started at a mean age of 7.4 (6-9) months. The results after 12 (n = 16), 24 (n = 12) and 30 (n = 8) months are presented. The mean height standard deviation score, SDS (range; Swedish standard), before therapy was -1.8 (-0.5 to -3.1) and the mean head circumference was -1.2 (-0.4 to -3.5). After 12, 24 and 30 months, the mean height SDS were 1.1 (-0.8 to -1.9), -0.9 (0 to -1.5) and -0.9 (0.1 to -1.5) and the mean head circumference SDS were -1.1 (0 to -2.5), -1.1 (0 to -2.2) and -1.2 (-0.5 to 2.0), respectively. During hGH-treatment, the children with DS thus gained height during the first year, and then followed the growth rate of healthy Swedish children. When compared to growth charts for children with DS the mean height of these children started at the fiftieth centile and reached the ninety-fifth centile after 24 months of treatment. Head circumference only slightly increased during the therapy, and not to the same extent as height. This indicates that small head circumference in DS is not only an effect of growth retardation, but also due to microcephaly. PMID- 8400722 TI - The development of learning difficulties in children with Down's syndrome. AB - This paper examines individual developmental profiles drawn from a series of longitudinal studies of cognitive development and problem-solving in children with Down's syndrome (DS) from birth to 11 years of age. These highlight the difficulties in explaining DS development in terms of a slowed-down version of normal development and illustrate how, from a very early age, developmental progress in DS is undermined by the children's failure to exercise and maintain existing skills and by their counter-productive approach to learning new skills. Developmental instability and inefficient learning were found to characterize performance on contingency detection tasks, on tests of object concept development, and on standardized intelligence tests. Implications of the findings for assessment, educational practice and developmental theory are discussed. PMID- 8400723 TI - Children's attitudes toward peers with intellectual disability. AB - The policy of inclusion (mainstreaming) of children with an intellectual disability in regular schools has raised questions about the extent to which 'true' integration is possible. One important aspect of integration is social acceptance by the regular class children. The purpose of this study was to determine the attitudes of children in primary and intermediate classrooms towards children with an intellectual disability housed in satellite classrooms at public schools. Teachers in the satellite classrooms completed a school integration questionnaire. Attitudes towards and the social distance afforded children in satellite classrooms were relatively positive across all children, especially girls. In particular, attitudes were more positive in schools which had more vigorous administrative policies concerning academic and social integration. The results are discussed in terms of current mainstreaming policies for children with intellectual disabilities. PMID- 8400724 TI - Teaching prereading skills to disabled children. AB - A Swedish survey suggests that less than 10% of children with Down's syndrome between 7 and 10 years of age are able to read a simple text. Children with Down's syndrome in a programme for early language intervention show clear progress in reading ability. The programme focuses on general language development, auditory and visual perception, and fine-motor ability. Prereading skills are stimulated in a systematic way such as identifying and discriminating sound-patterns at the level of the word, the syllable and the segments, and children are helped to categorize forms, to discriminate and understand pictures, to recognize the direction of reading, and to realize the grapheme-phoneme correspondence. PMID- 8400725 TI - A longitudinal study of cognitive skills and communication behaviours in children with Rett syndrome. AB - Changes in the cognitive, communicative and interactive development of a group of six girls with Rett syndrome (three younger and three older) were documented over a 3-year period. All six maintained a profound level of intellectual performance and a preintentional level of communication in which caregivers assigned meaning to the girls' limited behaviours. However, marked individual variation was noted in both cognitive and social interaction skills. The study supported the notion of an increased perception of social interactiveness by the caregivers over time. The development of means-end behaviour, in particular, seems closely linked with the measured increase in behaviours inferred as communicative. However, the results also show that factors such as cognitive level, physical status and educational intervention may be related to this perception. PMID- 8400726 TI - Freezing of embryos: early vs late stages. PMID- 8400727 TI - Freezing of preembryos: early vs late stages. PMID- 8400728 TI - Freezing of embryos: early vs late stages. PMID- 8400729 TI - Chemical removal of the outside of the zona pellucida of day 3 human embryos has no impact on implantation rate. AB - Two hundred eighteen consenting patients entered a randomized study of the application of chemical zona pellucida thinning on their day 3 embryos, prior to uterine transfer. Of those control patients (n = 108), whose embryos remained unmanipulated, 40 (37.0%) have ongoing/delivered pregnancies, while in the experimental group (n = 110), whose embryos had their zonae pellucidae chemically thinned, there are 49 patients (44.6%) who have ongoing/delivered pregnancies. Although this difference is not significant, clearly the application of this micromanipulative intervention has not been detrimental, and this bodes well for routine application of embryonic micromanipulation procedures in general. Certain patient subgroups were studied including older women, those with elevated basal follicle stimulating hormone levels, patients with embryos of differing zona thickness, and patients with embryos of differing uniformity of zona thickness. No significant influence of chemical removal of the outside of the zona on the implantation rate of embryos in any of these subgroups was observed other than a marginally significant (P = 0.095) improvement of implantation of embryos with less than 4.0 microns variation in zona thickness when chemical zona thinning was applied. Failure of chemical zona thinning to enhance human embryo implantation significantly, compared to assisted hatching by complete zona drilling, strongly suggests that the bilayered human zona pellucida needs to be fully breached, unlike that of the mouse. PMID- 8400730 TI - Effects of medium composition on murine and human blastocyst formation and hatching rate. AB - PURPOSE: Murine two-cell embryos (n = 5573) were cultured for 96 hr in human tubal fluid (HTF) medium (n = 2709) or alpha modification of minimum essential medium (MEM; n = 2864) through the hatched blastocyst stage from mid-1990 to mid 1991. An additional 373 embryos were cultured in MEM or HTF with 0, 1, 5, or 10 ng/ml E. coli endotoxin. A total of 17 patients had supernumerary embryos simultaneously cultured in HTF (n = 48) or MEM (n = 61). Additionally, pregnancy rates were compared for July to December 1990, when MEM was used as growth medium, and for July to December 1989, when HTF was used. RESULTS: Blastocyst formation was higher (P < 0.001) for murine embryos cultured in MEM (blasts = 95%) compared to HTF (blasts = 70%). When cultured with endotoxin, blastocyst formation was higher (P < 0.01) for embryos cultured in MEM compared with HTF for controls and at each endotoxin level. No difference in human blastocyst development was observed in HTF and MEM. However, more MEM-cultured blastocysts were cryopreserved (P < 0.05). There also was a lower spontaneous abortion rate and a higher multiple gestation rate when embryos were cultured in MEM. CONCLUSION: Thus, MEM may result in healthier blastocyst development, especially when culture conditions are substandard, although this is not an acceptable substitution for meticulous technique. PMID- 8400731 TI - The fate of embryos transferred into the uterus. AB - PURPOSE: The purpose of this study is to elucidate the fate of embryos transferred into the uterus. METHODS: Implantation rates were compared between synchronous and asynchronous intrauterine embryo transfers in mice. Then transferred embryos were recovered from the uterus and examined morphologically 24 hr after transfer. Moreover, the same transfer-recovery experiments were performed after ligation of the uterus two-thirds from the uterotubal junction immediately before transfer. RESULTS: The implantation rate was high (58.4% per embryo, 87.5% per recipient) when blastocysts were transferred into the uteri of recipients of pseudopregnant Day 4, but it was very low (0-2.4% per embryo, 0 8.3% per recipient) when blastocysts were transferred to recipients of pseudopregnant Day 2, or two-cell embryos into recipients of pseudopregnant Days 2 and 4. When transferred embryos were recovered from the uterus 24 hr after transfer, few embryos (1.4%) were recovered in asynchronous transfers, while 66.3% of the embryos were recovered in synchronous transfers. However, from recipients in which the lower portions of uteri were ligated, embryos at more advanced stages were recovered even in asynchronous transfers. CONCLUSIONS: These results suggest that discharge from the uterus is one of the major causes of the low implantation rate for asynchronous transfer. PMID- 8400732 TI - Slow and ultrarapid freezing of fully grown germinal vesicle-stage mouse oocytes: optimization of survival rate outweighed by defective blastocyst formation. AB - PURPOSE: The cryopreservation of mature metaphase II-stage mouse oocytes is associated with decreased fertilizability, spindle damage, and increased polyploidy. Therefore, we investigated the outcome of cryopreservation of immature germinal vesicle-stage mouse oocytes. METHODS: Oocytes were punctured from Graafian follicles in primed F1 hybrid mice and were then released into maturation medium containing the meiotic inhibitor dibutyryl cyclic AMP. Both slow and ultrarapid freezing protocols with dimethyl sulfoxide, 1,2-proponediol, or a mixture of both agents were tested. We recorded morphological survival rates, in vitro maturation rates, and two-cell and blastocyst formation rates. Each group of frozen oocytes was compared with both unfrozen germinal vesicle stage oocytes and metaphase II-stage oocytes. RESULTS: An optimal cryosurvival rate of 78% was reached after ultrarapid freezing with 3 M dimethyl sulfoxide followed by one-step dilution, but a decreased rate of two-cell formation was observed. Freezing with a combination of dimethyl sulfoxide and 1,2-propanediol did not improve this fertilization-decreasing effect. Very low cryosurvival rates after freezing with 1,2-propanediol indicated its inappropriateness for ultrarapid freezing of immature oocytes. The rates of in vitro maturation were equivalent for frozen-thawed and freshly collected germinal vesicle-stage oocytes, independent of the freezing protocol used. We report, nevertheless, as a general characteristic for both slow and ultrarapid freezing of fully grown germinal vesicle-stage oocytes, that the in vitro development up to the blastocyst stage is inhibited despite full nuclear maturation. CONCLUSION: We report that cryopreservation of immature germinal vesicle-stage oocytes is invariably associated with a low developmental capacity after fertilization. The rate of in vitro nuclear maturation did not equate with developmental competence. This in turn suggests the importance of cytoplasmic maturation for embryonic development. PMID- 8400733 TI - The significance of 17 beta-estradiol levels in highly responding women during ovulation induction in IVF treatment: its impact and prognostic value with respect to oocyte maturation and treatment outcome. PMID- 8400734 TI - A prospective study of echographic endometrial characteristics and pregnancy rates during hormonal replacement cycles. PMID- 8400735 TI - Supernumerous oocytes transferred in gamete intrafallopian transfer (GIFT). PMID- 8400736 TI - The relevance of endometrial thickness and echo patterns for the success of in vitro fertilization evaluated in 148 patients. PMID- 8400737 TI - A function test to assess the responsibility of oocyte and sperm quality in in vitro fertilization failure. PMID- 8400738 TI - Improved development of human embryos cultured on a Vero cell monolayer. PMID- 8400739 TI - Effect of fetal bovine serum of different gestational ages on mouse embryo growth and development. PMID- 8400740 TI - Effect of storage of Ham's F-10 medium on one-cell mouse embryo development in vitro. PMID- 8400741 TI - The development of a pressure area scoring system for critically ill patients: a pilot study. AB - At present no suitably sensitive scoring system is available to differentiate degrees of risk of developing pressure sores in critically ill people. The object of this study is to produce a tool that has been validated to identify and weight risk factors associated with critically ill patients. In a pilot study data were collected on 50 patients and analysed, using multiple regression analysis, to produce a set of weighted coefficients (C) suitable for use as a scoring system. The feasibility of the technique is demonstrated and five highly significant factors are described: adrenaline infusions (C = 14), noradrenaline infusions (C = 15), diabetes (C = 15), restricted movement (traction, post-operative pain, intra-aortic balloon pump and haemofiltration) (C = 8) and patients too unstable to turn (C = 17). All coefficients not equal to 0 with p < 0.05. Risk factors which may become significant in a larger study are identified. In order to develop the full tool a further multi-centre study is proposed. PMID- 8400742 TI - Teaching roles in critical care--the mentor and preceptor. AB - Mentorship and preceptorship have been terms used indiscriminately throughout the literature. The English National Board has used the term 'mentor' in all educational material and the term has slipped into current usage. Despite this, very little preparation and thought has been given to the role. Confusion has arisen as the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) Post Registration Education and Practice Project (PREPP 1990) proposals have outlined the need for a preceptor to support new practitioners to move confidently into a period of primary practice. This paper discusses the development of the mentor role and provides a definition of classical and formal mentoring. The preceptor role is viewed as one of the elements that enable the full development of mentorship. As a functional role, preceptorship can be clearly defined and utilised to support new staff into new areas of clinical practice. It crystallises the teaching, assessing and evaluative element of clinical nurse education and provides a firm foundation for the development of mentorship within the nursing profession. PMID- 8400743 TI - The near death experience. PMID- 8400744 TI - Occupational stress in neurosurgical nursing. AB - Occupational stress in nursing has been a popular topic for investigation. While many practice areas have been studied, the neurosurgical intensive care units (ICUs) and neuromedical/surgical units have largely been overlooked. Using interviews, this exploratory study examined aspects of neurosurgical nursing that were perceived as stressful by staff. Findings suggest that being exposed to life and death situations among young children, being short of essential resources, being on duty with too few staff and dealing with aggressive relatives constituted major stressful events. The frequency of occurrence of stressors was also investigated; working unsocial hours, being short of essential resources, being exposed to life and death situations in adults, and dealing with fluctuations in workload were cited as common happenings. Stressors perceived as having the most effect on work included being exposed to life and death situations among children and adults and being short of essential resources. Comments made by staff suggest that performance at work is adversely influenced by stress. While no neurosurgery-specific stressors were identified, the conclusion from this study is that intensely stressful events do occur in these ICUs and although such events may not happen often, they can adversely affect work performance, according to the staff involved. PMID- 8400745 TI - Gut feelings about gut feeding: enteral feeding for ventilated patients in a district general hospital. AB - This article looks at the role that nutrition plays in the maintenance of gut integrity and function in the critically ill patient. It refers to previous studies that have highlighted the relevance of facilitating the bacteriocidal function of gastric secretions as a means to minimize the development of nosocomial pneumonias in ventilated patients. A small trial was set up to investigate the merits and difficulties associated with two different enteral feeding regimes for ventilated patients in the intensive therapy unit (ITU) of a district general hospital. This project and its results are described and evaluated; conclusions and issues for practice are highlighted. PMID- 8400746 TI - Applying a model of quality assurance in intensive care. AB - At the moment many changes and new trends are being introduced in British health care. Perhaps amongst the most important of these changes are those aimed at introducing quality strategies into health care. In this paper the subject of quality assurance within intensive care unit settings is discussed, with an example of how one quality model might be implemented, Maxwell's model of quality (1984). PMID- 8400747 TI - A severe and fatal case of ethylene glycol poisoning. AB - A severe and fatal case of ethylene glycol (anti-freeze) poisoning in an unidentified vagrant man is reported and discussed. The case demonstrates the typical complications associated with this type of poisoning, in particular severe metabolic acidosis, acute renal failure and a deep comatose state. The importance of early diagnosis, aggressive treatment and intensive nursing care in order to optimize the chance of recovery are illustrated. PMID- 8400748 TI - The role of the nurse in ethical decision-making in intensive care units. AB - This paper focuses on the nurses' role in ethical decision-making in intensive care units (ICUs) of hospitals in the UK. There is a paucity of research on the topic, studies published are generally North American or Canadian in origin. The available literature suggests that although nurses are constantly at the intensive care patient's bedside, often making complex decisions about treatment and care, their involvement in ethical decision-making is limited. This paper provides a review of the literature on how ethical decisions are made and the influences on nurses' participation in this process. PMID- 8400749 TI - Comparison of three instruments for measuring patient anxiety in a coronary care unit. AB - This paper compares the State-Trait Anxiety Inventory (STAI), Hospital Anxiety and Depression Scale (HAD Scale) and a Linear Analogue Anxiety Scale (LAAS) for evaluating anxiety in patients with acute ischaemic heart disease. The instruments were examined for correlation, reliability and internal consistency. Strong associations were demonstrated at pre-test between the STAI and the other scales. Moderate coefficients between HAD-A and HAD-D/LAAS were also apparent. Lower correlations were found at post-test than at pre-test. At post-test, strong inter-correlations occurred for STAI/LAAS. The HAD Scale demonstrated high test retest reliability, while the STAI and LAAS were moderate in their reliability in this sample. The adequate correlation between the instruments suggest that each is a valid and appropriate measure of anxiety in this clinical sample. PMID- 8400750 TI - An overview of ventilator observations. AB - Nursing staff new to intensive care can often be apprehensive when observing, recording and interpreting ventilator observations. This review of ventilation and the necessary observations involved offers a description of the observations performed, with a problem-solving approach to cause, rationale and possible action that may be required. Calculations related to spontaneous and mechanical respiratory function are discussed, with recommendations for individual requirements. Explanation of available settings, demonstrating the functions and delivery methods of different ventilators is included, and a discussion on alarm settings and rationale. For the purpose of assessing the ventilatory support in use there is a guide to interpretation of arterial blood gases with suggestions and rationale for necessary changes of parameters. PMID- 8400751 TI - Laser Doppler flowmetry: a valid clinical tool? PMID- 8400752 TI - Effects of prostaglandin E1 or trimethaphan on local cerebral blood flow and carbon dioxide reactivity during cerebral aneurysm surgery. AB - Effects of prostaglandin E1 (PGE1) or trimethaphan (TMP) on local cerebral blood flow (LCBF) and carbon dioxide (CO2) reactivity were studied in 26 patient undergoing cerebral aneurysm surgery for subarachnoid hemorrhage (SAH) under isoflurane anesthesia. Measurement of LCBF was made using a thermal gradient blood-flow meter. Hypotension was induced with a continuous i.v. infusion of PGE1 or TMP. The dose of PGE1 or TMP was adjusted to maintain the MAP at about 70 mm Hg, and CO2 reactivity was studied during and after PGE1 or TMP administration. PaCO2 was changed by the controlled hyperventilation. CO2 reactivity was evaluated by the following formula: % delta LCBF/delta PaCO2% mm Hg. Hypotensive drugs were discontinued at the completion of aneurysm clipping. Mean arterial pressure (MAP) decreased immediately after the start of either PGE1 or TMP. Heart rate (HR) did not change significantly in either group. LCBF decrease at 30 min after start of TMP administration (p < 0.05 compared with preinfusion value), whereas LCBF was unchanged during PGE1 administration. Neither PGE1 nor TMP affected CO2 reactivity during cerebral aneurysm surgery, whereas CO2 reactivity showed a close correlation with presurgical neurological status: before, rs = 0.523, p < 0.01; during, rs = -0.794, p < 0.01; after, rs = -0.643, p < 0.01. Comparison of the outcomes after aneurysm surgery demonstrated no significant difference between groups; however, there was a close correlation between presurgical neurological status and outcome in both groups (rs = 0.829, p < 0.01). These results suggest that PGE1 is preferable to TMP in inducing hypotension for cerebral aneurysm surgery because this drug maintains both LCBF and CO2 reactivity. PMID- 8400753 TI - Laser-Doppler flowmetry in neurosurgery. AB - Laser-Doppler flowmetry (LDF) provides a reliable measurement of local cerebral blood flow (lCBF) as demonstrated by multiple validation studies. This article evaluates the clinical applications of LDF in neurosurgery. With the availability of modified probes, it is possible to carry out LDF monitoring of neurosurgical patients in the intensive care unit. Currently, there are multiple systemic and intracranial parameters that are interactive and separately monitored. A multi channeled digital data acquisition system allows these data to be compiled in a single computer environment for the interpretation of lCBF changes. Guidelines are suggested for the clinical use of LDF monitoring, and the technical features of monitoring including the interpretation of data are summarized. PMID- 8400754 TI - Airway pressure monitoring as an aid in the diagnosis of air embolism. AB - We designed a prospective study to compare the validity of airway pressure (AWP) monitoring with that of end-tidal CO2 (ETCO2) monitoring for early detection of air embolism. Subjects included 76 patients of both sexes who underwent neurosurgery in the sitting position. Anesthesia was maintained with O2, N2O, narcotics, pancuronium, and intermittent positive pressure ventilation (IPPV). Continuous monitoring was done of HR, ECG, intraarterial pressure, AWP, and ETCO2. A sudden and sustained decrease in ETCO2 during anesthesia in a hemodynamically stable patient was considered as a sign of air embolism. Concomitant changes in AWP and cardiovascular parameters were also recorded simultaneously. Onset time, stage of surgery, and duration of disturbances were recorded. At the same time, the chest was auscultated for any murmur. Aspiration of air through the CVP catheter was attempted for diagnosis and management of air embolism. ETCO2 monitoring detected 24 episodes (31.5%) of air embolism in 16 patients. We observed 10 episodes (13.1%) of tachycardia in nine patients and nine episodes (11.8%) of hypotension in eight of the 16 patients. Murmur was noted in four patients and air aspiration in six patients. Only six patients of the 16 had an increase in AWP along with the decrease in ETCO2. We conclude that AWP monitoring is neither a sensitive nor reliable indicator of air embolism. PMID- 8400755 TI - Anesthetic care during thiopental tests to evaluate epileptic patients for surgical therapy. AB - A thiopental test 2 weeks after insertion of intracranial electrodes may be used to evaluate patients with refractory epilepsy for surgical therapy. Barbiturates normally produce beta activity on the electroencephalogram. The absence of this response in a monitored brain region implies focal cerebral dysfunction. We describe a technique used to perform this test and the resultant morbidity. The thiopental test consists of intravenous injection of thiopental, 25 mg, every 30 s until either corneal reflexes are abolished, 1,000 mg of thiopental has been administered, or adverse events occur. In children, the dose is adjusted to approximately 0.3 mg/kg of thiopental every 20 s. A retrospective chart review was performed on 104 patients who underwent thiopental tests at the University of Pittsburgh Epilepsy Center. Records were systematically reviewed for thiopental dose, mean arterial blood pressure, heart rate, oxygen saturation in arterial blood, time to responsivity, need for airway intervention, and occurrence of nausea or vomiting. Thirty-six patients developed upper airway obstruction which required jaw lift maneuver, six patients were given 1,000 mg of thiopental without loss of corneal reflexes, and one patient briefly sustained an arterial saturation of 67%. Five patients exhibited electrographic seizures with clinical seizures evident in two patients. No permanent effects were evident in any patient as a consequence of the test. We conclude, with appropriate monitoring and personnel, that the thiopental test, as described, can be performed safely with acceptable morbidity. PMID- 8400756 TI - Clonidine premedication for craniotomy: effects on blood pressure and thiopentone dosage. AB - The purpose of this study was to determine whether oral clonidine premedication improves cardiovascular stability and/or reduces the requirements for drugs used to control systolic blood pressure (SBP) during elective craniotomies. We performed a double blind randomized trial involving 77 normotensive, ASA physical status I or II adults. Clonidine 4 micrograms/kg or placebo was given as oral premedication. The patient's mean SBP on the day before surgery was used as the baseline. SBP was controlled between the baseline and 30% below it (but not < 90 mm Hg). Anesthesia was induced with thiopentone, N2O, and fentanyl; maintenance was with N2O. Further doses of thiopentone were administered to control rises in SBP until skin incision. After skin incision trimethaphan (TMP) was used to control the SBP and isoflurane only added if TMP was insufficient. Isoflurane was discontinued as soon as SBP control allowed. The induction dose of thiopentone was the same in both groups but subsequent usage for blood pressure control was significantly lower in the clonidine group. There were no differences in trimethaphan dose, or the incidence and duration of isoflurane use. The clonidine group had lower SBP on arrival in the operating room, preinduction, and postintubation. There were no differences in mean "intraoperative" SBP, their coefficients of variation, or recovery room values. Two subgroups were analyzed, based on the study groups mean age and baseline SBP. Three-way analysis of variance revealed that the blood pressure effects of clonidine were almost entirely confined to patients older than 45 years. Baseline SBP had no independent effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400757 TI - Rate of cerebrospinal fluid formation, resistance to reabsorption of cerebrospinal fluid, brain tissue water content, and electroencephalogram during desflurane anesthesia in dogs. AB - Intracranial pressure (ICP) has been shown to increase dramatically during desflurane anesthesia, possibly as a result in part of an increase in the rate of cerebrospinal fluid (CSF) formation (Vf) or a decrease in the rate of CSF reabsorption. To examine this phenomenon, I designed a study to measure Vf, resistance to reabsorption of CSF (Ra), brain tissue water content, and the electroencephalographic activity (EEG) during desflurane anesthesia in dogs. Vf and Ra were determined using ventriculocisternal perfusion of mock CSF labeled with blue dextran. EEG activity was determined using aperiodic analysis. At the end of the study, brain tissue water contents of gray and white matter were determined by dry/wet weight ratios. Eighteen dogs were allocated into three groups. Group 1 (n = 6) was examined at five experimental conditions during normocapnia; group 2 (n = 6) was examined at five experimental conditions during hypocapnia. The experimental conditions for groups 1 and 2 were (a) baseline (halothane 0.5-1.0% inspired plus thiopental 12 mg.kg-1 i.v. given over 15 min followed by i.v. infusion at 12 mg.kg-1 x h-1), (b) 0.5 MAC (3.5 +/- 0.1% expired) and (c) 1.0 MAC (7.0 +/- 0.1% expired) desflurane at normal CSF pressure, and (d) and (e) 0.5 and 1.0 MAC desflurane at increased CSF pressure (> 30 cm H2O). Group 3 (n = 6), the control group, was examined over the same time period as groups 1 and 2. In the control group, desflurane was not administered; instead, the baseline condition (i.e., halothane plus thiopental) was maintained throughout the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400758 TI - Cerebral blood flow in a rat glioma model during halothane anesthesia. AB - Little is known about the influence of infiltrating gliomas on the responsivity of the cerebral circulation to anesthetic agents. Therefore we designed a study to address this issue. Male Fischer 344 rats were assigned to two tumor groups and one sham group. In the two tumor groups, glioma cells were stereotactically injected into the right striatum; animals in the sham group were injected with sterile culture medium only. Either 12 or 16 days after injection, the rats were anesthetized with 1 MAC halothane in 40% O2/balance N2. Local and remote regional cerebral blood flow was then determined using 14C-iodoantipyrine autoradiography. Physiologic values (PaCO2, PaO2, pHa, mean arterial pressure, and rectal temperature) were similar for both tumor and sham groups. Tumor volume was relatively small (cross-sectional diameter = 2-3 mm), and there was no evidence of midline shift in coronal tissue sections. Blood flow within the tumor was substantially reduced relative to adjacent structures (e.g., tumor = 88 +/- 10 ml/100 g/min; adjacent caudate = 161 +/- 23 ml/100 g/min). There were no significant differences between the tumor and sham groups for regional blood flow values in histologically normal tissue in either the injected or contralateral hemispheres. We conclude that this model of brain neoplasia shows no evidence of either local or remote changes in the cerebrovascular responsibility of normal tissue to volatile anesthesia. PMID- 8400759 TI - Combined therapy with dizocilpine and levemopamil does not reduce ischemic neuronal injury in rats. AB - Excessive intracellular accumulation of calcium has been postulated to result in ischemic neuronal death. Reduction of intracellular calcium entry should therefore be expected to reduce ischemic neuronal injury. Two pathways through which extracellular calcium ions can enter neurons are voltage-sensitive and N methyl-D-aspartate receptor-linked cation pores. Combined blockade of both these types of channels might be more effective in reducing intracellular calcium accumulation than the blockade of either channel alone. We therefore evaluated the cerebroprotective effects of dizocilpine, an N-methyl-D-aspartate receptor antagonist, and levemopamil, a phenylalkylamine calcium channel blocker, administered singly or in combination, in a model of forebrain ischemia in the rat. Four groups of rats (n = 8 each) were studied. In the first group, dizocilpine, 5 mg/kg, was administered before ischemia. In the second group, levemopamil, 5 mg/kg, was given both preischemia and 2 h postischemia. In the third group, both dizocilpine (5 mg/kg) and levemopamil (5 mg/kg) were given preischemia and levemopamil (5 mg/kg) was given postischemia. The control group received saline placebo. The rats were subjected to forebrain ischemia by bilateral carotid artery occlusion for 10 min with simultaneous hypotension to 35 mm Hg. Neuronal injury was evaluated 3 days after ischemia. Dizocilpine reduced postischemia neuronal injury in the ventral hippocampus (p = 0.045). Levemopamil and the combination of levemopamil and dizocilpine did not protect neurons from ischemic injury. The present study does not provide support for the strategy of combined therapy with dizocilpine (administered before ischemia) and levemopamil (administered before and after ischemia) to protect neurons from injury produced by severe incomplete forebrain ischemia. PMID- 8400760 TI - Cerebral monitoring during cardiopulmonary bypass in children. AB - Although cerebral monitoring during CPB remains primarily investigational, recent data support its clinical utility. In particular, it is cerebral metabolic monitoring that provides meaningful information in terms of preparing the brain for dhCPB and dhCA. Cerebral blood flow or cerebral blood flow velocity monitoring is less beneficial due to the presence of luxuriant cerebral blood flow at deep hypothermic temperatures. Conventional temperature monitoring can be improved upon by adding jugular venous oxygen saturation monitoring to satisfy the primary goal of cerebral protection--uniform cerebral cooling and metabolic suppression. Although online measures of cerebral cellular metabolism are not widely available, early experience with near infrared technology suggests that it is a feasible and reliable monitor of cerebral metabolic activity and is likely to represent an important noninvasive continuous monitor in the near future. CMRO2 recovery data have suggested that cerebral metabolic suppression is more severe the longer the period of dhCA. Cerebral protection strategies, such as intermittent cerebral perfusion have demonstrated less metabolic suppression of dhCA in animal models and are currently undergoing clinical evaluation in our institution. Finally, the postoperative period remains a high-risk period for neurologic injury because temperatures are normothermic, cardiac output is reduced, cerebral autoregulation is impaired, and management strategies, such as hyperventilation, are commonly used to increase pulmonary blood flow with little knowledge on its effects on cerebral perfusion. PMID- 8400761 TI - Hippocampal tin, aluminum and zinc in Alzheimer's disease. AB - The use of inductively coupled plasma source mass spectrometry (ICP-MS) for multi element analysis has led to the observation, in two separate studies, of increased blood tin in Alzheimer's disease (AD). We have therefore applied the technique of ICP-MS to hippocampal tissues obtained post-mortem from patients with AD and from controls. There was no significant difference in tin concentrations in AD. There were increased concentrations of aluminum and silicon, and reduced concentrations of zinc and selenium. It is postulated that displacement of hippocampal zinc by heavy metals may be important in producing clinical memory disturbance. However, analysis of the CA1 region, rather than of the dentate gyrus, would have been preferable. PMID- 8400762 TI - Some biochemical aspects of the protective effect of strontium chloride on gamma irradiated rats. AB - The effect of treatment with SrCl2 (10 mg 100 g-1) on rats 15 min prior to whole body gamma-irradiation (7.5 Gy) was studied. The hazardous effects of irradiation were greatly corrected in the treated group. The hyperglycemic effect and liver glycogen accumulation in the untreated group decreased to normal level. The enzymatic activities of serum alkaline phosphatase, alanine and aspartate aminotransferases, and lactate dehydrogenase were greatly affected, showing insignificant changes in the treated group of animals. Life span calculated on 50% survival was also significantly elongated by 36.3%. These results show the potentiality of SrCl2 as a radioprotective agent. A proposed mechanism is discussed. PMID- 8400763 TI - Effect of stress, adrenalectomy and changes in glutathione metabolism on rat kidney metallothionein content: comparison with liver metallothionein. AB - Eighteen hours of immobilization stress, accompanied by food and water deprivation, increased liver metallothionein (MT) but decreased kidney MT levels. Food and water deprivation alone had a significant effect only on liver MT levels. In contrast, stress and food and water deprivation increased both liver and kidney lipid peroxidation levels, indicating that the relationship between MT and lipid peroxidation levels (an index of free radical production) is unclear. Adrenalectomy increased both liver and kidney MT levels in basal conditions, whereas the administration of corticosterone in the drinking water completely reversed the effect of adrenalectomy, indicating an inhibitory role of glucocorticoids on MT regulation in both tissues. Changes in glutathione (GSH) metabolism produced significant effects on kidney MT levels. Thus, the administration of buthionine sulfoximine, an inhibitor of GSH synthesis, decreased kidney GSH and increased kidney MT content, suggesting that increased cysteine pools because of decreased GSH synthesis might increase kidney MT levels through an undetermined mechanism as it appears to be the case in the liver. However, attempts to increase kidney MT levels by the administration of cysteine or GSH were unsuccessful, in contrast to what is known for the liver. The present results suggest that there are similarities but also substantial differences between liver and kidney MT regulation in these experimental conditions. PMID- 8400764 TI - Osteotoxicity of cadmium and lead in HOS TE 85 and ROS 17/2.8 cells: relation to metallothionein induction and mitochondrial binding. AB - Epidemiological, experimental and clinical data indicate that cadmium and lead are osteotoxins in man and other species. The relative sensitivities of a clonal human osteosarcoma cell line (HOS TE 85) and a clonal rat osteosarcoma cell line (ROS 17.28) to the cytotoxic effects of cadmium and lead were tested in serum free media without added growth factors. The rat osteosarcoma cells were more sensitive to cadmium with cytotoxicity and inhibition of proliferation at 0.25 versus 0.75 and 1.0 mumol l-1 cadmium, respectively, for human osteosarcoma cell lines. The lower sensitivity to cadmium of human osteosarcoma cells is attributed, at least partly, to induction of metallothionein synthesis by cadmium and zinc in this cell line; in the rat osteosarcoma cell line, they do not induce metallothionein synthesis. Human osteosarcoma cells were more sensitive than rat osteosarcoma cells to lead with inhibition (IC50) of proliferation at 4 mumol l-1 lead and cytotoxicity at 20 versus 6 and over 20 mumol l-1 lead, respectively, for these variables in rat osteosarcoma cells. Both cell lines attained the highest lead concentration in the 15,000 x g (mitochondrial) fraction. The lead in the mitochondrial, microsomal, nuclear and cytosolic fractions of the human cell line did not decrease during 24 h post-washout. Binding of lead was much less stable in the less sensitive rat cells, with 50-100% loss of mitochondrial, microsomal and nuclear lead during 24 h post-washout. PMID- 8400765 TI - Purification and chemical characterization of staphyloferrin B, a hydrophilic siderophore from staphylococci. AB - This paper describes the chemical characterization of staphyloferrin B, a new complexone type siderophore isolated from low iron cultures of Staphylococcus hyicus DSM 20459. Purification of the very hydrophilic metabolite was achieved by anion exchange high performance liquid chromatography HPLC. Mass spectrometry showed a molecular mass of 448 amu. Hydrolysis with 8 M HCl revealed the presence of L-2,3-diaminopropionic acid, citrate, ethylenediamine and succinic semialdehyde. The connections between the four building blocks were determined by two-dimensional nuclear magnetic resonance measurements. UV/Vis and circular dichroism spectra are consistent with the proposed structure, which could also be confirmed by precursor feeding. The siderophore activity of staphyloferrin B was demonstrated by iron transport measurements. PMID- 8400766 TI - [Relation of hypothyroidism and deficiency of kidney yang]. AB - 32 cases of hypothyroidism caused by various factors were treated for one year with Chinese medicinal herbs preparation "Shen Lu tablet" (SLT) to warm and reinforce the Kidney Yang. 34 normal persons were studied as a control group. After treatment with SLT, the clinical symptoms of hypothyroidism were markedly improved. Average serum concentration of total T3, T4 increased significantly from 67.06 +/- 4.81 and 3.29 +/- 0.42 before treatment to 120.50 +/- 6.34 ng/dl and 6.31 +/- 0.38 micrograms/dl, respectively (P < 0.001). Serum levels of TSH decreased evidently from 20.81 +/- 2.78 before therapy to 3.32 +/- 0.61 ng/ml (P < 0.001). Before treatment with SLT, hypothyroidism group had higher erythrocyte sodium content ([Na]Rbc) than that of normal group. The permeability of the cell membrane oMNaos and the activity of sodium pump oKNaos in intact erythrocytes were markedly lower in the treated group than that in the normal group. In hypothyroidism patients treated by SLT the [Na]Rbc significantly dropped, and the oNNaos and oKNaos were significantly raised when compared with those before treatment, P < 0.001. It is suggested that hypothyroidism was closely related with Deficiency of Kidney Yang and energy metabolism. PMID- 8400767 TI - [Doppler sphygmogram of uneven pulse and spectral changes of tongue]. AB - The volume of blood flow (F), its velocity (V) and the internal diameter (D) of patients with uneven pulse (PUP) were determined and compared with control. RESULTS: The F, V and D of PUP were significantly less than those of control respectively (P < 0.001). It provided objective basis for Blood Stasis. Furthermore, the value of spectrum of tongue of PUP were measured. Values of red and orange ray of PUP were less, while those of green and blue ray were more than those of normal subjects respectively. The value of purple ray were negative correlated with F and V. The values of red and orange ray were correlated with F and V. It revealed that uneven pulse and purple tongue existed simultaneously in majority. Combination of the two signs was in favour of the diagnosis of blood Stasis. PMID- 8400768 TI - [Effect of tonifying kidney on compliability of the aged]. AB - The clinical study of 80 aged individuals divided into treated group (Group I) and normal control (Group II) was carried out. According to the symptoms of Kidney Deficiency the aged was subdivided into Kidney Deficiency group (KDG) and non-Kidney Deficiency group. A semi-quantitative method and some indexes about compliability were used to evaluate the symptoms. RESULTS: After treatment the main symptoms of Kidney Deficiency such as lumbago, cold limbs, fatigue, cold aversion, feverish sensation in chest, palms and soles, etc. were significantly alleviated (P < 0.05) and the speed of reaction (SR) that reflects NS function in Group I and its KDG was also significantly improved. After exercise test, the ultrasono-cardiogram showed that in KDG of Group I, the ascending range of heart rate (HR) lessened during exercise and recovered more rapid than that before treatment. The difference was significant (P < 0.05). The HR was also significant different between the KDG groups of Group I and II during resting, exercise and recovering status (P < 0.05). Furthermore, after treatment the level of salival progesterone of the aged female was significantly. The determination of bone mineral content showed that in KDG after treatment, the density of ulnar line was remarkably elevated. These results showed that through tonifying Kidney and replenishing Qi could improve the functions of heart, brain, bone and endocrine systems of the aged persons, therefore improve their compliability. PMID- 8400769 TI - [Metrological analysis for efficacy of acupuncture on angina pectoris]. AB - Acupuncture (Acupoints: Neiguan P6, Shenmen H7, Shaohai H3 or auriculo-points: Heart, Shenmen) was administrated once or 7 times in a week on 40 patients with stable type of angina pectoris. The effect was assessed quantitatively or semi quantitatively according to the extent, area, frequency, duration of attack, the time of attack during exercise, and the vanishing of suffering after exercise. Just after one performance of acupuncture, 15 patients' angina pectoris were significantly alleviated (P < 0.001) both in degree and area. After 7 times of acupuncture 10 patients' angina pectoris were not only significantly alleviated both in extent and area, but also in frequency and duration of attack. 15 patients were randomized to an acupuncture, non-acupuncture or acupuncture at non acupoints (ANA) in a single blind design. The time from the beginning of exercise to the anginal attack in active acupuncture group was longer than that in non acupuncture or ANA group (P < 0.01), but they were similar (P > 0.05) in both non acupuncture group and ANA group. The time from the end of exercise to the disappearance of angina pectoris in acupuncture group was shorter than that in the other two groups (P < 0.05). PMID- 8400770 TI - [Effects of combined therapy of traditional Chinese and Western medicine on C reactive protein in post-debridement patients]. AB - Closely monitoring whether the secondary infection in the patients of post debridement occurred or not and appropriately treating these patients were the important ways to reduce the incidence of infection. Through estimating the level of the serum C-reactive protein (CRP) as the monitoring index of infection, dynamically observed the effect of the combined traditional Chinese and Western medicine therapy (TCM-WM) on CRP after debridement, as was compared with the effect of the Western medicine therapy (WM) group in which only the WM was administrated. The result showed that the levels of CRP decreased in both TCM-WM and WM group on 4th day after the operation, but the level of CRP in former group was lower than that in latter one, the difference was very significant (P < 0.001). So that, it was assumed that TCM-WM significantly excelled the WM on affecting the level of CRP and reducing the incidence of infection. It was suggested that CRP could be used as an effective and objective index to determine whether the secondary infection has happened and to assess the efficacy of some drugs. PMID- 8400771 TI - [75 infantile palsy children treated with acupuncture, acupressure and functional training]. AB - In treating infantile cerebral palsy (CP), 75 CP children were treated with a comprehensive meridian therapy including scalp and body acupuncture, acu-point injection and auriculo-point stimulation, supplemented with acu-pressure and massage, and functional training. A minimum of 10 times of treatment within twenty days, and a maximum of 120 times within a year was performed. The effect of the treatment was evaluated by appraising the children's performance of physical exercise and their social adaptability. The intelligence quotient (IQ) of 30 sick children that had been treated for 60 times (6 courses) was compared prior to and after treatment. It indicates that the treatment yielded a very positive improvement in the children's physical capability and an increase of their intelligence. PMID- 8400772 TI - [Enhancing effect of jian pi jin dan on immune functions of normal and cyclophosphamide induced immunosuppressed mice]. AB - By means of normal and cyclophosphamide (CY) injected NIH mice, the effect of Jian Pi Jin Dan on immuno-modulation was studied which could treat the "Gan" disease effectively in TCM. RESULTS: Markedly improved the level of serum lysozyme, enhance the phagocytosis of abdominal macrophage. The proliferation of spleen T, B cells, the production of interleukin-1 (IL-1) by macrophage and of interleukin-2 (IL-2) by T cells in normal and CY injected mice were also enhanced. Furthermore, it was able to restore the weight, spleen and thymus index of CY injected mice. This prescription can not only reinforce Spleen, but also regulate Liver, complying to the children's physiological and pathological characteristics. PMID- 8400773 TI - [Comparison of anti-inflammatory, analgesic activities, anaphylactogenicity and acute toxicity between bee venom and its peptides]. AB - Bee venom 1.0-2.0 mg/kg and bee venom peptides 1.0-2.0 mg/kg inhibited several inflammatory processes, such as ear swelling induced by xylene in mice, edema produced by injecting 1% carrageenin 0.1 ml beneath the plantar surface of hind paw in rats and showed a marked analgesic action induced by the hot plate and potassium antimony tartrate. Bee venom peptides had a markedly more effective action as compared with bee venom itself. The anaphylactogenicity of bee venom peptides was apparently milder than that of bee venom. The LD50 of bee venom ip in mice and bee venom peptides was 7.4 mg/kg and 7.9 mg/kg respectively. PMID- 8400774 TI - [Effect of ligustrazine on isolated myocardial ischemic reperfusion injury in rats]. AB - In recent years, it is believed by some scholars that the injury of myocardial ischemic reperfusion is correlated to the thromboxane A2 (TXA2) released by platelets. In order to explore that whether the myocardial and hemangio endothelial cells participate in the TXA2 production during the process of reperfusion, the modified Langendorff method was used to establish the model of reperfusing the isolated rat heart. On the other hand, this experiment was also intended to observe the effect of ligustrazine on the injury of myocardial ischemic reperfusion. The results revealed that the level of thromboxane B2 (metabolite of TXA2) and lactic dehydrogenase (LDH) in coronary sinus reflux fluid increased during the process of reperfusion, while the level of 6-keto-PGF1 alpha in the same fluid relatively decreased (P < 0.05). The ratio of TXB2/6-keto PGF1 alpha was raised. The ligustrazine inhibited the release of TXB2 and LDH, but promoted the production of 6-keto-PGF1 alpha (P < 0.05). The results also proved that the myocardial and hemangio-endothelial cells could synthesize TXA2, and the amount of TXA2 released increased during the reperfusion of ischemic myocardium, which was likely to be the major factor of the injury of ischemic myocardial reperfusion. Ligustrazine plays an important role in protecting the myocardium. PMID- 8400775 TI - [Some problems of clinical application on pediatric external treatment]. PMID- 8400776 TI - [Research on basic theory of traditional Chinese medicine]. PMID- 8400777 TI - [Effect of Chinese materia medica on natural killer cell, lymphokine-activated killer cell and activity of interleukin 2]. PMID- 8400778 TI - Is there a place for barter within nurse education? PMID- 8400779 TI - Continence care: a forgotten specialty. PMID- 8400780 TI - Postoperative pain in the elective surgical patient. AB - Postoperative pain in the elective surgical patient needs effective management to maximize comfort. This article describes ways to ensure that pain is given priority in patient care by acknowledging that it is subjective and differs between individuals. PMID- 8400781 TI - The nurse's role in caring for people with learning disabilities. AB - Nurses caring for people with learning disabilities are facing role confusion and virtual extinction as a consequence of changes in care provision. This article draws from the results of a working party established to clarify their role and offers a model which can provide comprehensive and quality health care. PMID- 8400782 TI - Working with schizophrenic clients and their families. AB - This article presents a history of schizophrenia family work. It describes research carried out into the subject and how and why the knowledge gained is now being disseminated to mental health nurses. PMID- 8400783 TI - Care in theater of a patient undergoing tonsillectomy. PMID- 8400784 TI - Employing quantitative and qualitative methods in one study. AB - There is an apparent lack of epistemological rigour when quantitative and qualitative methods are combined in the same study, because they reflect opposing positivist and interpretive perspectives. When and how to use methodological pluralism is discussed in this article. PMID- 8400785 TI - Warning: some courses can damage your education. PMID- 8400786 TI - Nurse-aid management of ear and nose emergencies: 1. AB - Ear, nose and throat emergencies cause a great deal of discomfort and distress and in certain cases may compromise safety. They therefore demand skilful intervention on the part of the nurse aider. In this article, the first of a two part series, the nurse-aid management of ear problems will be described. The next article in the series will concentrate on the nose. PMID- 8400787 TI - Screening the over-75s: 2. AB - Although Government guidelines assume that screening of the over-75s will have beneficial effects, the content of the assessment is not clear. This article, the second in a two-part series, presents a tool for functional assessment. PMID- 8400788 TI - The clostridia and biotechnology. PMID- 8400789 TI - Molecular biology and genetics of substrate utilization in clostridia. PMID- 8400790 TI - Xylan and cellulose utilization by the clostridia. PMID- 8400791 TI - Molecular biology of nitrogen fixation in the clostridia. PMID- 8400792 TI - Properties of acid- and solvent-forming enzymes of Clostridia. PMID- 8400793 TI - [Effects of short-term application of recombinant human growth hormone on urea production rate in patients in the early postoperative phase]. AB - OBJECTIVE: To determine the effect of short-term recombinant human growth hormone (rhGH) administration on urea production rate (UPR), N balance and aminograms. DESIGN: Prospective, double-blind placebo-controlled study. SETTING: Intensive care unit of a University Hospital. PATIENTS: 20 adult patients after major abdominal surgery. INTERVENTIONS: Postoperative substitution of rhGH (Saizen, Serono, Aubonne, CH) in a dose of 0.15 IE/kg BW for 3 days. The serum levels of hGH, insulin-like growth factor-1 (IGF-1), ACTH and cortisol were measured as well as the amino acids in plasma. The degree of catabolism was calculated according to Woolfson's formula, which is based on the UPR, and by calculation of the cumulative N balance. RESULTS: With exception of proline, the plasma amino acids between the groups receiving active substance and total parenteral nutrition (TPN) and those receiving placebo and TPN did not differ significantly. Neither was there a significant difference between the groups for any other parameter measured. The UPR and IGF-1 levels showed only a tendency towards higher values as compared with the placebo group (UPR verum group, values in g/day: 1st measurement, 29.8 +/- 16.7; 2nd measurement, 28.3 +/- 17.7; 3rd measurement, 32.1 +/- 19.1; 4th measurement, 33.1 +/- 21.2. UPR placebo, values in g/day; 1st measurement, 32.6 +/- 23.9; 2nd measurement, 30.8 +/- 17.9; 3rd measurement 41.6 +/- 28.7; 4th measurement, 47.3 +/- 29.5. IGF-1 verum group, values in nmol/l; 1st measurement, 25.7 +/- 19.2; 2nd measurement, 44.8 +/- 23; 3rd measurement, 52.4 +/- 30; 4th measurement, 54.3 +/- 20. IGF-1 placebo, values in nmol/l; 1st measurement: 22.9 +/- 11.7; 2nd measurement, 37.0 +/- 19.4; 3rd measurement, 38.4 +/- 21.4; 4th measurement, 40.0 +/- 23.0). The ACTH-cortisol axis was only slightly depressed in the group receiving active substance. CONCLUSIONS: We conclude that short-term rhGH administration over 3 days is not capable of significantly reducing the UPR in postoperative patients, but we cannot exclude a significant difference between rhGH group and placebo after a longer administration period. PMID- 8400794 TI - [Thrombogenicity of central venous catheters: comparison of two polyurethane catheters]. AB - BACKGROUND: Thrombosis represents a possible catheter-related risk. Beside many endogenous and exogenous factors, the material as well as its condition do have a decisive influence on thrombogenicity. One factor of thrombogenicity is the surface of the catheter. Therefore, two different types of polyurethane catheters were compared according to their surface structure by electron microscopy. Furthermore, an in vivo testing should confirm a possible, different thrombogenicity. MATERIAL AND METHODS: A model with a single aperture at the catheter tip and a model with multiple apertures were tested. We compared the electron-microscopic pictures of specific areas which were expected to cause problems. Thrombogenicity in the vena cava inferior was tested in 20 rabbits. For statistical evaluation the Wilcoxon U test was used. RESULTS: Polyurethane catheters have a smooth and homogeneous surface inside and outside of the catheter. However, there are many production-related, qualitative differences in the morphology of the apertures. The model with multiple apertures and a closed catheter tip turned out to have a much more irregular surface compared to the model with a single aperture. In vivo testing showed that each catheter causes thrombosis, independent of the type. The catheter with multiple apertures proved to have no advantages as far as thrombogenicity is concerned; catheter-specific complications of this type are possible. Also, atraumatic insertion by means of Seldinger's technique is not possible using a catheter with a closed tip. CONCLUSION: The catheter with multiple apertures does not have any advantages to the catheter with a single aperture. PMID- 8400795 TI - [Detection of rare rhesus antigens Cx and Ew using monoclonal and polyclonal antibodies in routine laboratory tests]. AB - Two cases are described in whom rare antigens of the rhesus blood group system could be found by absent or very weak reaction with monoclonal antisera and strong reaction with polyclonal reagents. All 3 monoclonal anti-E sera showed no reaction with the Ew antigen, while monoclonal anti-C sera had different reactions with the Cx antigen. PMID- 8400796 TI - [Removal of additive solutions?]. PMID- 8400797 TI - [Monitoring in hemodilution]. AB - Normovolaemic haemodilution is an established part within the 'Concept of Autologous Transfusion'. According to the mechanisms to compensate for the dilution-induced anaemia, monitoring of haemodilution has to consider (1) maintenance of normovolaemia; (2) stability of the cardio-vascular system and of a normal pulmonary function; (3) an adequate myocardial oxygen supply. (1) Normovolaemia: Under routine clinical conditions normovolaemia is controlled by close monitoring of fluid balance (considering surgical blood loss, diuresis, and insensible perspiration). If the expected blood loss is > 2.0 litres, additional monitoring of the central venous pressure appears to be reasonable. It is not a single value of the central venous pressure (CVP) but rather its time-course that allows conclusions on changes of intravascular volume. (2) Cardio-vascular and pulmonary function: Pulmonary function is easily controlled by intermittent arterial blood gas analysis. Non-invasive and discontinuous or invasive and continuous blood pressure recording, respectively, are routinely used for monitoring of cardiovascular function. Heart rate together with the time-course of the CVP give additional information on the cardio-vascular system. Central venous oxygen saturation is only a minor substitute for mixed venous oxygen saturation; however, its changes with time make it possible to draw conclusions on global haemodynamics and total body oxygen supply. However, in situations of extreme haemodilution--as in Jehova's witnesses--a pulmonary artery catheter has to be used for monitoring the cardio-vascular system as well as bulk oxygen parameters. (3) Myocardial oxygen supply: Monitoring for myocardial ischaemia is routinely performed by ECG. It is both the number and the kinds of leads chosen that give adequate information.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400798 TI - [Monitoring blood coagulation in larger liver operations]. AB - Bleeding is causally related to about 50% of postoperative deaths following liver resection. Main factors contributing to increased perioperative bleeding in liver surgery include surgical trauma, reduced activity of clotting factors and inhibitors due to impaired hepatic synthesis, low platelet count and poor platelet function as well as impaired clearance of activated clotting factors by the reticuloendothelial system of the liver (Kupffer cells). Hemostasis may be further impaired by transfusion of blood components, since citrate added for conservation is not adequately metabolized by the failing liver. Surgical bleeding leads to a loss of pro- and anticoagulatory factors as well as to activation of coagulation. Finally, hyperfibrinolysis induced by release of tissue plasminogen activator (t-PA, primary hyperfibrinolysis) and disseminated coagulation (secondary hyperfibrinolysis) contribute to increased bleeding. Therefore early diagnosis and treatment of coagulation disorders is of paramount importance during liver surgery. Screening parameters of hemostasis and fibrinolysis should be available on a 24-hour basis in centers performing liver surgery. Screening for disorders of secondary hemostasis includes evaluation of prothrombin time (PT), activated partial thromboplastin time (aPTT), fibrinogen concentration and the activity of the most important inhibitor, antithrombin III (AT III). Thrombelastography is the leading method for diagnosis of hyperfibrinolysis, which can also be assessed by determination of D-dimer, fibrinogen and fibrin degradation products. Evaluation of primary hemostasis is frequently restricted to platelet count, which is only a rough parameter. In contrast, measurement of in vitro bleeding time and volume enables repeated quantification of platelet function in patients with impaired hemostasis. PMID- 8400799 TI - Perioperative respiratory monitoring of oxygen transport. AB - Oximetry nowadays is understood as the in vitro measurement of O2 saturation (sO2, %) and hemoglobin (Hb) derivatives (%) using 4-7 wavelengths (CO- and Hem oximeters). Pulse oximeters, using only 2 wavelengths, are designed for the continuous noninvasive measurement of the arterial partial O2 saturation (psO2, %) in vivo. Light-emitting diodes allow light to pass through the peripheral site of measurement with red and infrared light to enable a distinction between oxygenated and deoxygenated hemoglobin during a recorded pulse wave. In case of physiological concentrations of Hb derivatives the determination of psO2 is performed with clinically relevant accuracy of +/- 2-3%. However, at carboxyhemoglobin (COHb) concentrations above normal, under normoxia as well as under hypoxia the accuracy of measurement varies considerably among the instruments from different manufacturers. In the case of elevated methemoglobin (MetHb) concentrations, the situation is completely different. With increasing cMetHb, the psO2 is still the value required, but success depends on the concentration of MetHb: under normoxia psO2 is increasingly underestimated, whereas under hypoxia increasing overestimation must be anticipated. Provided there is a constant Hb concentration, knowledge of the initial sO2, and absence of the derivatives COHb and MetHb as well as of severe perfusion disorders, pulse oximetry is suitable for perioperative respiratory monitoring of oxygen transport. PMID- 8400800 TI - [Dependence of O2 uptake on O2 transport--a myth or reality?]. AB - According to the concept of supply dependence of oxygen uptake, an improvement of tissue oxygenation in critically ill patients may be obtained by an increase in systemic oxygen transport. A critical review of available clinical data suggests that a positive correlation between calculated oxygen uptake and oxygen delivery does not necessarily imply a 'pathological' dependence of both parameters. Possible pitfalls in the interpretation of results, like mathematical coupling of data, alterations in oxygen demand due to variations in sedation and the calorigenic effect of catecholamines, are discussed. PMID- 8400801 TI - Investigations of antisense oligonucleotides targeted against bcl-2 RNAs. AB - Expression of the bcl-2 gene becomes deregulated in many non-Hodgkin lymphomas as the result of t(14;18) chromosomal translocations. Because bcl-2 regulates the survival of cells, and because its over-expression is associated with cellular resistance to killing by chemotherapeutic drugs and gamma-irradiation, this gene and its mRNA and protein products represent ideal targets for designing novel therapeutic strategies for the treatment of cancer. Here we describe the effects of an 18-mer phosphodiester oligonucleotide that is complementary to the first 6 codons of the bcl-2 mRNA's open reading frame. When tested for inhibition of in vitro protein synthesis using RNAse-H-supplemented reticulocyte lysates and RNA prepared by in vitro transcription of a human bcl-2 cDNA, the bcl-2 antisense (AS) oligomer completely abolished Bcl-2 protein production at 10 microM, but had no effect on the in vitro translation of a chicken bcl-2 RNA that contained three mismatches relative to the oligomer binding site on the human bcl-2 RNA. A control 18-mer having the same base composition as the AS oligomer but with scrambled order (SC) was not inhibitory. Addition of AS and SC oligomers to cultures of a NIH-3T3 fibroblast cell line that had been stably infected with a recombinant retrovirus containing the same human bcl-2 cDNA used for in vitro transcription/translation experiments revealed concentration-dependent reductions in the relative levels of the 26-kD human Bcl-2 protein (as determined by immunoblotting) by the AS but not by the SC oligomer. Similar results were obtained when AS and SC oligomers were applied to a t(14;18)-containing lymphoma cell line SU-DHL-4 that was cultured in low-serum media. When used at 200 microM, the bcl-2 AS oligomer produced 84-95% reductions in Bcl-2 protein levels in SU DHL-4 cells but had relatively little effect on the levels of other mitochondrial control proteins, suggesting that the inhibitory effects were specific. Treatment of SU-DHL-4 cells with AS oligomer lead to essentially complete loss of bcl-2 mRNA from cells within 1 day of addition to cultures, but presumably because of the long half-life of the Bcl-2 protein (approximately 14 h), commensurate reductions in Bcl-2 protein levels did not occur until 3 days.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8400802 TI - Evaluation of the use of antisense tRNA(met) as an inhibitor for eukaryotic protein synthesis. AB - We attempted to explore the use of antisense RNAs against tRNA as an inhibitor of eukaryotic protein synthesis. For this purpose, antisense RNA against the 5'-end half of the initiator tRNA of wheat germ was synthesized, and its effect on translation of the Brome mosaic virus mRNA was investigated in a wheat germ cell free system. When the antisense RNA against the 5'-end half of the initiator tRNA including the anticodon sequence was added at the concentration of 8 microM to the cell-free system, protein synthesis was completely inhibited. This inhibitory effect could be suppressed by the addition of wheat germ tRNA. In contrast, sense and control RNA showed slight inhibitory effects, which were not, however, suppressed by wheat germ tRNA. The antisense tRNA formed a double-stranded RNA duplex with the target methionine tRNA in the wheat germ extract which became resistant to ribonuclease treatment. These experiments suggest that antisense tRNA could be utilized for control of tRNA functions and to block protein synthesis. PMID- 8400803 TI - Systematic analysis of antisense RNA inhibition of myosin II heavy-chain gene expression in Dictyostelium discoideum. AB - The powerful potential of antisense nucleotide inhibition of gene expression is presently being exploited in many biological systems. Although use of the technique is widespread, little is known about the variables that contribute to experimental success. We sought to define those variables that affect inhibition of myosin II heavy-chain (MIIHC) gene expression by stable nuclear-derived antisense RNAs in Dictyostelium discoideum. Different fragments of the MIIHC gene cloned in the antisense orientation into several transformation vectors were introduced into cells, and the accumulation of MIIHC protein and mRNA was examined. Inhibition of expression ranged from slight to virtually complete, and depended only on the specific gene fragment used to generate the antisense RNA. Fragments of the 3' end of the gene were the most effective and resulted in almost complete inhibition, whereas 5' fragments gave very little reduction. The severity of the morphological phenotypes associated with MIIHC depletion reflected the different levels of MIIHC in the transformants. Important implications for the design of antisense vectors suggested by these results are discussed. PMID- 8400804 TI - Co-suppression of Dictyostelium discoideum myosin II heavy-chain gene expression by a sense orientation transcript. AB - We have previously demonstrated effective inhibition of Dictyostelium discoideum myosin heavy chain (MIIHC) gene expression after introduction of an antisense gene on a transformation vector into cells. We now report that transcription of a sense orientation MIIHC gene has a similar, though not as dramatic, effect on expression from the endogenous gene. Cells transformed with a sense fragment of the MIIHC gene are defective in cytokinesis and become giant and multinucleated when growing on a surface. This morphological phenotype is correlated with decreased expression of the endogenous MIIHC gene. Sense transformants accumulate less MIIHC protein and mRNA than control transformed cells, concomitant with accumulation of a vector-derived sense transcript. Expression of the introduced sequences seems to be required for the sense effect. The possible causes of this surprising phenomenon are discussed. PMID- 8400805 TI - Sequence-specific DNA damage using iodine-125-labeled antisense oligonucleotides. AB - A procedure is described that cleaves single-stranded DNA with sequence specificity. This process involves attaching a DNA damaging agent to an oligonucleotide. This oligonucleotide delivers the DNA damaging agent, iodine 125, to a specific DNA sequence by complementary hybridization. 5-[125I]Iodo-2 deoxycytidine 5'-triphosphate was enzymatically incorporated into an oligonucleotide that was designed to hybridize to a single-stranded DNA target. 125I decays by electron capture and causes breaks in the target DNA. These breaks were observed on a DNA sequencing gel. After 22 days of exposure to the 125I labeled oligonucleotide, significant damage was observed within 1 to 2 bases of the expected site of hybridization. Densitometry showed that after 48 days the amount of damage had approximately doubled. This method facilitates easy design and testing of oligonucleotides that could potentially be used to inactivate gene expression in a wide variety of organisms. PMID- 8400806 TI - International conference on Nucleic Acid Medical Applications, NAMA conference. PMID- 8400807 TI - Transient intra-areal axons in developing cat visual cortex. AB - Transient axons reaching the medialmost part of area 17 were demonstrated with anterogradely transported biocytin injected in the dorsal part of the lateral gyrus in kittens during the first and second postnatal weeks. The axons decreased in number during the third postnatal week and were only exceptionally found thereafter. Computer-aided reconstructions from serial sections demonstrated axons with different degrees of complexity. The most complex ones were found at postnatal days 7-9 and were characterized by multiple branches terminating with growth cones in the white matter. Characteristically, endings of axons that entered the cortex remained confined to the infragranular layers V and VI. A few axons entered the supragranular layers. Transient axons terminated with different endings, which may indicate different stages of maturation. A few, possibly permanent, axons were still found in the medial part of area 17 at the end of the first, and during the second postnatal months; they arborized widely in the infragranular layers, and modestly or not at all supragranularly. PMID- 8400808 TI - Effects of concomitant cholinergic and adrenergic stimulation on learning and memory performance by young and aged monkeys. AB - The beneficial effects of physostigmine and other centrally acting AChE inhibitors on learning and memory processes have been documented in several studies. Some of these compounds are currently being examined for their potential in the treatment of Alzheimer's disease. However, the ability to employ this class of agents alone is limited by the potential for debilitating and dangerous side effects. In addition, a growing body of evidence also suggests a role for a number of other transmitter systems in this disorder. Agents such as clonidine that address adrenergic deficits have recently been demonstrated to enhance memory performance in monkeys. Clonidine also inhibits the function of cholinergic neurons in specific brain regions and reduces certain side effects of physostigmine. This evidence provided the impetus to evaluate a combination of physostigmine and clonidine in a learning and memory paradigm in monkeys. Seven young adult and three aged macaque monkeys performing a delayed matching-to sample task received regimens of increasing doses of clonidine and physostigmine on separate occasions to determine the optimal dose of each agent in terms of enhanced memory performance. The optimal dose of clonidine was combined with a series of physostigmine doses and performance was compared to that using the two drugs alone. The best combination regimen of clonidine and physostigmine was more effective than either drug alone in enhancing memory performance. Part of the benefit may have been due to the ability of the animals to tolerate significantly higher doses of physostigmine in the combination regimen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400809 TI - Topographical variation of the human primary cortices: implications for neuroimaging, brain mapping, and neurobiology. AB - The relationships of the "primary" cytoarchitectonic neocortical fields, 17, 41, 3b, and 4 (Brodmann areas), to salient topographic landmarks have been reconstructed from serial histological sections in 20 human cerebral hemispheres (10 brains). Each of these architectonic fields is found to bear a characteristic relationship to a set of enframing anatomic landmarks, in particular, gyri, fissures, and sulci, that can be readily defined by MRI. Two classes of variability were found characteristic, at least to some extent, of each of the fields. Class 1 variability--variability that is not predictable from visible landmarks--was typical of the polar and for the cuneal and lingual extracalcarine distributions of field 17 and the distribution of field 4 upon the paracentral lobule. Class 2 variability--variability that is closely predictable from visible landmarks--is seen in the marked interindividual or interhemispheric variation in size or shape of a field and was found to be prominent for all four fields. Because of the prominence of class 2 variability, direct reference to the landmarks that frame these fields may be expected to be a more reliable basis for functional mapping than reference to a template or stereotactic coordinate-based system of reference to a standard or idealized brain. PMID- 8400810 TI - Three-dimensional quantitative analysis of hemispheric asymmetry in the human superior temporal region. AB - The recent observations of overall symmetry of the caudal infrasylvian region by Steinmetz et al. (1990) and Witelson and Kigar (1991, 1992) diverge from earlier findings of leftward asymmetry in this region (Geschwind and Levitsky, 1968; Galaburda et al., 1987; Larsen et al., 1989). To address this inconsistency, we measured the entire infrasylvian surface posterior to Heschl's gyrus from coronal magnetic resonance images of 10 young, normal, right-handed subjects. Computer models were constructed by tracing contours of this region and then interpolating a 3D triangle mesh between each pair of adjacent contours. Measurements of these models showed no significant directional asymmetry. The same contour set was used to obtain measurements with a conventional algorithm that does not interpolate a surface between contours. The results obtained with the second method showed significant leftward asymmetry. These results suggest that in some cases, unbalanced distortions due to folding differences of the hemispheres are sufficient to obtain spurious findings of left-right asymmetry. This supports the claim of Steinmetz and Witelson that leftward asymmetry is restricted to the temporal bank of the caudal infrasylvian surface, and is balanced by rightward asymmetry of the parietal bank. PMID- 8400811 TI - Cortical Discover Award: David Fitzpatrick, associate professor, Department of Neurobiology, Duke University School of Medicine, Durham, North Carolina. PMID- 8400812 TI - Cortical Discover Award: Charles D. Gilbert, professor, the Rockefeller University, New York, New York. PMID- 8400813 TI - Cortical Explorer Award: Gregg H. Recanzone, Laboratory of Sensorimotor Research, National Institute of Health, Bethesda, Maryland. PMID- 8400814 TI - Cortical Scholar Award: Dr. Bradley L. Schlaggar, the Salk Institute for Biological Studies, La Jolla, California. PMID- 8400815 TI - Sciatic nerve stimulation induces hypotension but not renal or lumbar sympathoinhibition in hypertensive Dahl rats. AB - Sustained reductions in arterial pressure and sympathetic nerve activity occur after prolonged sciatic nerve stimulation in spontaneously hypertensive and pre hypertensive Dahl salt-sensitive rats whereas these responses are not observed in renal hypertensive or Dahl resistant rats. These observations suggest that the development of post-stimulation hypotension and sympathoinhibition may be related to the genetic predisposition for hypertension rather than to the increased level of arterial pressure. However, it is not known whether the magnitude of the post stimulation blood pressure and sympathetic nerve responses are influenced by the increased level of arterial pressure in addition to the genetic predisposition to hypertension. In the present study, we sought to determine if sustained sciatic nerve stimulation induces post-stimulation hypotension in hypertensive Dahl sensitive (DS) rats. For this purpose, mean arterial pressure (MAP), heart rate (HR), renal (RSNA) and lumbar (LSNA) sympathetic nerve activity were recorded during and after sciatic nerve stimulation in hypertensive DS rats (n = 17) fed an 8.0% NaCl diet for 7-8 weeks. Sciatic nerve stimulation increased HR (control, 443 +/- 10 b.p.m.; stimulation, 487 +/- 8 b.p.m.; p < 0.05) and tended to increase MAP, RSNA and LSNA. Two hours after stimulation, MAP was reduced (control 145 +/- 5 mmHg; recovery, 124 +/- 8 mmHg; p < 0.01) from control values. In contrast, RSNA and HR remained unchanged whereas LSNA was increased (69 +/- 20%; p < 0.05) from control values 120 min after stimulation. MAP, HR and RSNA were unchanged from control values during and for 2 h after sham stimulation in eight DS rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400816 TI - Impaired respiratory sinus arrhythmia with paradoxically normal Valsalva ratio indicates combined cardiovagal and peripheral adrenergic failure. AB - Combined reductions of the heart rate responses to deep breathing (HRDB) and the Valsalva ratio (VR) are used as a standard test of cardiovagal function. We observed that some patients had marked reduction of HRDB with a paradoxically normal VR. The mechanism of this paradox was evaluated. We found that these patients had evidence of peripheral adrenergic and postganglionic sudomotor impairment, including an excessive fall in blood pressure on upright tilt. Since VR depends on heart rate responses to blood pressure excursions, we evaluated the phases of the Valsalva manoeuvre and the ensuing heart rate responses in these patients and compared the responses to patients with generalized autonomic failure (Group II) and controls (Group III). Group I patients when compared with controls, had a larger phase II fall in blood pressure and normal phase IV resulting in a significantly enhanced baroreflex stimulus. These patients had a normal heart rate increment but 75% failed to develop reflex bradycardia. We conclude that the paradoxical VR is due to the exaggerated blood pressure swing due in turn to adrenergic failure. When both HRDB and VR are reduced as in Group II, there may be the additional impairment of cardiac adrenergic failure. PMID- 8400817 TI - A comparison of cardiovascular reflex tests and spectral analysis of heart rate variability in healthy subjects. AB - Determination of whether results of cardiovascular reflex tests and spectral analysis of heart rate variability are age dependent and whether there is correlation between results of both, cardiovascular reflex tests (the Valsalva manoeuvre, deep breathing test, handgrip test, cold face stimulus test, orthostatic test) and spectral analysis of heart rate variability were performed on 83 healthy volunteers of both genders, aged 21 to 70 years. We found that results of all heart rate based tests and results of spectral analysis decreased with aging, while results of blood pressure based tests did not. Parasympathetic activity predominated in younger subjects, while in older subjects sympathetic activity was dominant. Valsalva, deep breathing, and orthostatic ratios correlated with integrals of amplitude spectra in the standing posture and deep breathing and cold face stimulus ratios with integrals of amplitude spectra in the supine posture, whereas blood pressure changes during handgrip and orthostatic test did not correlate with integrals of the amplitude spectra. These findings suggest that tests based on heart rate may be more sensitive than tests based on blood pressure changes. This study supports the use of spectral analysis as an additional clinical test of autonomic nervous system function and stresses the importance of age in the evaluation of the results of autonomic nervous system function testing. PMID- 8400818 TI - Vasomotor reflexes in the fingertip skin of patients with type 1 diabetes mellitus and leprosy. AB - Fingertip skin blood flow was measured by laser Doppler flowmetry (as LDflux) under environmental conditions promoting vasodilation in Scottish patients with diabetes mellitus and Indian patients with leprosy. The reflex control of fingertip blood flow was assessed by measuring the reduction in LDflux induced by deep inspiratory gasp (IG) and cold challenge (CC) of immersing the contralateral hand in cold water. The uncomplicated diabetic patients showed normal vasomotor reflexes and an increased, though non significant, LDflux level (p < 0.06). The patients with diabetic neuropathy had resting LDflux levels significantly less than the uncomplicated group and also had substantial impairment of both IG and CC reflexes. Those with retinopathy (but no clinically apparent neuropathy) had LDflux within the normal range, but they showed minor evidence of impairment of the vasomotor reflexes. The uncomplicated newly registered leprosy patients had reduced LDflux and substantial impairment of CC reflexes. These changes were more marked in newly registered leprosy patients with clinical evidence of neuropathy. Leprosy patients with long-standing neuropathy requiring orthopaedic treatment had LDfluxes so greatly reduced that measurement of vasomotor reflexes was not practicable. The CC reflex was more severely affected than the IG reflex and more frequently absent in leprosy patients, possibly because of associated sensory neuropathy affecting the afferent limb of this response. Thus laser Doppler flowmetry can detect impairment of reflex control of fingertip blood flow in both diabetes mellitus and leprosy, but there are functional differences in the pattern of autonomic impairment between the diseases, suggesting differences in the pathogenesis of nerve damage. PMID- 8400819 TI - The effects of postural change and exercise on renal haemodynamics in familial dysautonomia. AB - Cardiovascular instability is a prominent manifestation of familial dysautonomia [FD] while renal insufficiency occurs in a large number of adult FD patients. To determine if there was a causative relationship, renal artery blood flow velocity using Doppler technology, was recorded and the ratio of the peak systolic velocity (point A) to the end diastolic velocity (point B) was calculated. The A/B ratio was assessed in response to change of position and exercise, and was correlated with renal function, heart rate and systemic blood pressure. Studies were performed in 54 FD patients with a mean age of 24 years +/- 9.8 years, and 20 controls, with a mean age of 24.7 +/- 7.6 years. In the supine position, the mean A/B ratios were not significantly different, but FD subjects had a significantly higher mean blood pressure and heart rate than controls. When erect and post exercise, the mean A/B ratios in FD subjects were significantly higher than controls, p = 0.0004 and p = 0.0001, respectively. In contrast to controls, when FD subjects were standing erect and post exercise, mean blood pressure decreased significantly without a significant change in heart rate. When FD subjects were divided into two groups based on their creatinine clearance value, the group with the lower creatinine clearances had a significantly greater fall in diastolic pressure when they moved from the supine to the erect position. Our results indicate that noninvasive Doppler techniques are helpful in detecting changes in renal blood flow in subjects with familial dysautonomia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400820 TI - Long-term variability and reproducibility of resting human muscle nerve sympathetic activity at rest, as reassessed after a decade. AB - Human muscle nerve sympathetic activity measured by microneurography during supine rest is known to vary considerably between healthy subjects, whereas in a given individual the level of muscle nerve sympathetic activity is stable over weeks and months. To further characterize long-term variability or reproducibility microneurographic recordings of muscle nerve sympathetic activity were performed in 15 healthy, normotensive subjects (mean age 51 years) who had undergone the same procedure between 10 and 14 years earlier (mean 12 years). The range of muscle nerve sympathetic activity was 9-59 in the first and 13-61 bursts/min in the second recording. Subjects maintained the level of muscle nerve sympathetic activity displayed previously, although with a slight but significant tendency to a higher outflow with increasing age. It is concluded that muscle nerve sympathetic activity is characterized by large inter-individual differences and strong intra-individual reproducibility over many years, with a tendency to increase with age. The age relationship is only in a minor part responsible for the variability, the cause of which remains unexplained. Because of the marked difference between individuals, strict normality criteria are difficult to define when comparing groups of subjects. There remains the risk of either obtaining spurious differences or obscuring a true abnormality. This is unlikely to apply when results in individual subjects are compared. PMID- 8400821 TI - The biology of glycoconjugates. PMID- 8400822 TI - A method for glycoconjugate synthesis. AB - 7-Formylheptyl glycosides of 2-acetamido-2-deoxy-beta-D-glucopyranose and O-alpha L-rhamnopyranosyl-(1-->3)-O-alpha-L-rhamnopyranose were synthesized and were coupled by reductive amination to bovine serum albumin and aminopropyl glass, respectively. PMID- 8400823 TI - Synthesis of polymeric neoglycoconjugates based on N-substituted polyacrylamides. AB - Several types of polymeric glycoconjugates, N-substituted polyacrylamides, have been synthesized by the reaction of activated polymers with omega aminoalkylglycosides: (i) (carbohydrate-spacer)n-polyacrylamide, 'pseudopolysaccharides'; (ii) (carbohydrate-spacer)n-phosphatidylethanolaminem polyacrylamide, neoglycolipids, derivatives of phosphatidylethanolamine; (iii) (carbohydrate-spacer)n-biotin-polyacrylamide, biotinylated probes; (iv) (carbohydrate-spacer)n-polyacrylamide-(macroporous glass), affinity sorbents based on macroporous glass, covalently coated with polyacrylamide. An almost quantitative yield in the conjunction reaction makes it possible to insert in the conjugate a predetermined quantity of the ligand(s). Pseudopolysaccharides proved to be a suitable form of antigen for activation of polystyrene and poly(vinyl chloride) plates (ELISA) and nitrocellulose membranes (dot blot), being advantageous over traditional neoglycoproteins. Polyvalent glycolipids insert well in biological membranes: their physical properties, particularly solubility, can be changed in a desired direction. Biotinylated derivatives were used as probes for detection and analysis of lectins. PMID- 8400824 TI - Purification and properties of the alpha-3/4-L-fucosyltransferase released into the culture medium during the growth of the human A431 epidermoid carcinoma cell line. AB - A soluble alpha-3/4-fucosyltransferase secreted into the growth medium of the human A431 epidermoid carcinoma cell line has been purified 700,000 fold by a series of steps involving chromatography on Phenyl Sepharose 4B, CM-Sephadex C-50 and GDP-hexanolamine Sepharose 4B. The untreated spent culture medium transferred almost ten times more fucose to the subterminal N-acetylglucosamine residue in the Type 1 (Gal beta 1-3GlcNAc) disaccharide than to the subterminal sugar in the Type 2 (Gal beta 1-4GlcNAc) disaccharide; the relative activity with these two substrates remained virtually unchanged throughout the purification procedure. At no stage was any alpha-3-fucosyltransferase species acting solely on N acetylglucosamine residues in Type 2 chains separated from the bulk of the alpha 3/4-fucosyltransferase activity. The purified enzyme preparation showed insignificant activity with glycoprotein substrates having N-linked oligosaccharide chains with terminal Type 2 sequences but transferred fucose to a mucin-type glycoprotein with O-linked oligosaccharide chains with terminal Type 1 structures. Lactose was a poor substrate but the activity of the enzyme was influenced by the presence of substituents on the terminal beta-galactosyl residue and 2'-fucosyllactose was almost as good an acceptor as the Type 1 disaccharide. The properties of the purified enzyme with regard to specificity, divalent cation requirements, pH optimum, and M(r), closely resembled those of the Lewis-blood-group gene associated alpha-3/4-fucosyltransferase isolated from human milk. PMID- 8400825 TI - UDP-galactose:lactosylceramide alpha-galactosyltransferase activity in human placenta. AB - The activity of UDP-Gal:LacCer galactosyltransferase in human placenta was studied by using crude homogenate and Triton CF-54 extract as the source of enzyme. Transfer of galactose to lactosylceramide was optimal in the presence of 0.1% Triton CF-54 and Mn2+ at pH 6.3, and the reaction product was susceptible to alpha-galactosidase. PMID- 8400826 TI - Effects of cyclodextrins on the hydrolysis of ganglioside GM1 by acid beta galactosidases. AB - The hydrolysis of ganglioside GM1 by acid beta-galactosidases was greatly enhanced by the inclusion of heptakis(2,6-di-O-methyl)-beta-cyclodextrin or alpha cyclodextrin in the assay mixture. The other cyclodextrins tested were not effective. The extent of stimulation by these cyclodextrins was relatively smaller than those by taurodeoxycholate and taurochenodeoxycholate. However, it is suggested that stimulation by bile salts may be partly a reflection of the detergent effects of bile salts on GM1 and partly a reflection of the interaction between bile salts and the enzyme itself. On the other hand, the stimulation by the cyclodextrins seems to correlate to the formation of an inclusion complex between GM1 and cyclodextrin without enzyme protein interaction. PMID- 8400827 TI - Structural assessment of beta-glucuronidase carbohydrate chains by lectin affinity chromatography. AB - Rat liver beta-glucuronidase was studied by sequential lectin affinity chromatography. beta-Glucuronidase glycopeptides were obtained by extensive Pronase digestion followed by N-[14C]acetylation and desialylation by neuraminidase treatment. According to the distribution of the radioactivity in the various fractions obtained by chromatography on different lectins, and on the assumption that all glycopeptides were acetylated to the same specific radioactivity, a relative distribution of glycan structure types is proposed. The presence of complex biantennary and oligomannose type glycans (56.8% and 42.7%, respectively) was indicated by Concanavalin A-Sepharose chromatography. Ulex europaeus agglutinin-agarose chromatography revealed the presence of alpha(1 3)linked fucose in some of the complex biantennary type glycans (16.6% of the total glycopeptides). Wheat germ agglutinin chromatography indicated that the minority (0.5%) were hybrid or poly (N-acetyllactosamine) type glycans. Furthermore, the absence of O-glycans, tri-, tetra- and bisected biantennary type glycans was demonstrated by analysis of Concanavalin A-Sepharose unbound fraction by chromatography on immobilized soybean agglutinin, Ricinus communis agglutinin and Phaseolus vulgaris erythroagglutinin. PMID- 8400828 TI - Identification of capillaries in sections from skeletal muscle by use of lectins and monoclonal antibodies reacting with histo-blood group ABH antigens. AB - This study was performed to evaluate the application of different lectins and monoclonal antibodies against ABH antigens to detect and characterize carbohydrate structures in capillaries of skeletal muscle from humans and laboratory animals. Blood group specific lectins (Griffonia simplicifolia, Griffonia simplicifolia isolectin B4, Lotus tetragonolobus, Ulex europaeus, and Dolichos biflorus) and monoclonal antibodies reacting with histo-blood group carbohydrate antigens belonging to type 1 (Le(a)) and type 2 (H, A and Le(y)) chains were used as histological markers for capillaries in sections from skeletal muscle. The material consisted of 20 human masseter muscle biopsies from individuals with known blood types: (eight blood group O, nine blood group A, two blood group B, and one blood group AB) and masseter muscles specimens from different laboratory animals (mouse, rat, rabbit, cat, dog, pig, cow, and macaca monkey). Unfixed sections and an avidin alkaline phosphatase method were used to visualize the specific reaction. Ulex lectin stained capillaries in all human biopsies either strongly or moderately. Strong muscle capillary reaction was observed in biopsies from O, B and AB individuals while capillaries from A individuals were only moderately stained. Griffonia simplicifolia marked capillaries in A, B, and AB individuals and Griffonia simplicifolia isolectin B4 stained capillaries in muscle biopsies from B and AB donors. Dolichos biflorus was a weak marker of muscle capillaries from A individuals. Only capillaries from O individuals were stained with the antibody against H type 2. Capillary reaction was not observed with the other antibodies used. Girffonia simplicifolia was an excellent marker for capillaries in mouse muscle while Griffonia simplicifolia isolectin B4 is recommended for rat muscles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8400829 TI - Prospects for NMR of large proteins. AB - During the last decade, solution structures of many small proteins have been solved by NMR. The size of proteins that are being analyzed by NMR seems to increase steadily. Protein structures up to 18 kD have been solved so far, and spectra of proteins up to 30 kD have been assigned. Thus, NMR emerges as an attractive technique, in particular for structural studies of proteins that cannot by crystallized. However, the application of the technology is limited by relaxation properties of the proteins. If relaxation would only be determined by Stokes-Einstein-type rotational diffusion, the effects of the molecular size on relaxation properties of proteins and thus on the performance of multi dimensional multiple-resonance experiments could readily be estimated. From this perspective, solving two- or three-fold larger structures seems possible. However, most larger proteins exhibit serious line broadening due to aggregation or other still unknown effects. Sample conditioning to minimize these effects is presently the challenge in the work with large proteins. PMID- 8400830 TI - Conformational analysis of alpha-D-Fuc-(1-->4)-beta-D-GlcNAc-OMe. One-dimensional transient NOE experiments and Metropolis Monte Carlo simulations. AB - One-dimensional transient NOE build-up curves were measured for the synthetic disaccharide alpha-D-Fuc-(1-->4)-beta-D-GlcNAc 1 utilizing Gaussian shaped pulses. Simulated build-up curves from Metropolis Monte Carlo simulations were compared to the experimental data. Disaccharide 1 is structurally related to methyl beta-D-maltoside in that it also contains an alpha-(1-->4) linkage, and it has the same configuration of groups around the glycosidic linkage. Analysis of NOEs in methyl beta-D-maltoside is restricted to those observed upon selective excitation of H1' because of severe spectral overlap. The situation is different in 1 where 1H-NMR signals are well separated. Several interglycosidic NOEs were observed. The corresponding build-up curves allowed an accurate determination of the conformational preferences at the glycosidic linkage in 1. Comparison of experimental and theoretical NOE build-up curves showed clearly that rigid minimum-energy models cannot account for the experimental data. The best fit of experimental NOE build-up curves was obtained with theoretical curves from a 2 x 10(6) step Metropolis Monte Carlo simulation with the temperature parameter set at 1000 K. Finally, it was observed that only the interglycosidic NOE H5'/H6-pro S significantly depends upon varying conformation distributions at the alpha-(1- >4)-glycosidic linkage, induced by choosing different temperature parameters for the Metropolis Monte Carlo simulations. PMID- 8400831 TI - Heteronuclear NMR studies of 13C-labeled yeast cell wall beta-glucan oligosaccharides. AB - The structures of uniformly 13C-labeled beta-glucan octa- and undeca oligosaccharides enzymatically prepared by the yeast cell wall glucanosyl transferase of Candida albicans were characterized by using a combination of HCCH COSY, HCCH-TOCSY, and HMBC experiments. The oligosaccharide structures indicate that the cell wall glucanosyl transferase cleaves two glucosyl units from the reducing end of the initial linear beta(1-->3) penta-oligosaccharide and subsequently transfers the remainder to another oligosaccharide at the nonreducing end via a beta(1-->6) linkage. These results indicate that the combined action of cell wall glucanase and glucanosyl transferase activities could not only introduce intrachain beta(1-->6) linkages within a single glucan strand, but also result in cross-linking of two initially separate glucan strands with concurrent introduction of intrachain beta(1-->6) linkages. Since isolated fungal membranes only synthesize linear beta(1-->3) glucan strands, wall associated enzymes probably participate in the assembly of the final wall glucan structure during cell growth and division. PMID- 8400832 TI - Binomial frequency response to non-binomial pulse sequences for efficient water suppression. AB - This article reports on the use of short-hard pulse and spin-lock pulse combinations giving a binomial-like frequency response for the measurement of NMR spectra in aqueous solutions of quite dilute samples. The pulse sequence proposed provides excellent water suppression and does not introduce any linear or higher order phase errors. Application to the measurement of 2D NOESY data of a 0.25 mM solution of a double-stranded DNA fragment is presented. PMID- 8400833 TI - A simple and sensitive experiment for measurement of JCC couplings between backbone carbonyl and methyl carbons in isotopically enriched proteins. AB - A simple 2D difference experiment is described that allows quantitative measurement of 13C-13C J couplings between backbone carbonyl and side-chain carbons. Precise 3JCC values were measured from data recorded in just 2 h for a 1 mM solution of the 20-kD complex between the protein calmodulin and a 26-residue synthetic peptide. The J couplings aid in determining the chi 1 angles of valine, isoleucine and threonine residues, and in making stereospecific assignments of the Val C gamma methyl groups. Error analysis indicates that the uncertainty in the derived J couplings is generally less than ca. 0.3 Hz. PMID- 8400834 TI - The relationship between quitting smoking and changes in drinking in World War II veteran twins. AB - Smoking has been associated with the intake of alcohol, and abstinence from tobacco has been hypothesized to increase alcohol intake. There is no consensus, however, on the extent of changes in alcohol intake following withdrawal from nicotine. The current study of a large cohort (n = 5,510) of World War II male veteran twins examined, prospectively, changes in drinking in relation to changes in smoking over a 16-year time period in late adulthood. Self-reported histories of smoking habits obtained from two surveys, administered in 1967-1969 and in 1983-1985, allowed the classification of subjects into continuing smokers (n = 1,441), quitters (n = 1,760), and continuing nonsmokers (n = 2,309). Analyses of the change in alcohol consumption over the period of follow-up, after adjustment for baseline consumption, socioeconomic variables, and health status, found: (1) a significant but moderate increase in alcohol consumption in quitters, due to an increase in wine consumption; (2) a significant marked increase in alcohol consumption in continuing smokers due to an increase in consumption of spirits; and (3) no significant change in alcohol consumption in continuing nonsmokers. These findings indicate that smoking cessation, a behavior change that promotes health, is associated with an increase in alcohol consumption, which may have other adverse long-term effects on health in this cohort. PMID- 8400835 TI - Nicotine and caffeine use in cocaine-dependent individuals. AB - Nicotine and caffeine use in 87 cocaine-dependent persons seeking treatment at an outpatient clinic were compared to use of those substances in a matched general population sample (n = 78). The prevalence of cigarette smoking was significantly greater in the cocaine-dependent sample (75% vs. 22%). Within the cocaine dependent sample, smokers were younger, less educated, employed in lesser skilled jobs, and reported an earlier onset and more frequent use of cocaine. The prevalence of caffeine use was significantly less in the cocaine-dependent group (68% vs. 83%), although, among caffeine users, the cocaine group drank significantly more caffeinated beverages per day than matched controls (4.9 vs. 3.3). Interestingly, regular caffeine use was associated with less frequent cocaine use within the cocaine-dependent sample. To our knowledge, this study is the first to report on prevalence of smoking and caffeine use among cocaine dependent individuals, and suggests that use of these other substances may influence the onset and pattern of cocaine use. PMID- 8400836 TI - An empirical description of phases of maintenance following treatment for alcohol dependence. AB - This article reports two longitudinal prospective studies conducted sequentially to describe participants' maintenance of abstinence experiences up to 36 months posttreatment. Study 1 reports responses of 102 subjects who maintained alcohol abstinence following treatment and who were assessed for duration and intensity of symptoms associated with postacute withdrawal at baseline, 3, 6, and 12 months. A decreasing linear trend of symptoms was found as time of alcohol abstinence increased. Study 2 obtained interview and survey data from 23 successful abstainers from Study 1 at 12, 18, and 36 months posttreatment to describe coping strategies, perceptions of relapse risk, extent and duration of "aftercare," and substitute addictions as these phenomena developed and changed over time. Multiple occasions of data collection supported the description of three phases of maintenance: symptom stabilization/management accompanied by a cognitive paradigm shift, distancing self from alcohol-dependent behavior, and normalization of life processes. PMID- 8400837 TI - Discriminative features of alcoholics classified according to family history. AB - By means of discriminant analysis, the frequently proposed distinction of alcoholics according to family history of alcoholism was tested. The most powerful discriminative factors were dysfunctional attitudes, some particular personality characteristics, and perceived parental rearing patterns. The results lend support to the assumption to regard alcoholics with a positive family history of alcoholism as a homogenous subgroup characterized by a specific etiopathogenesis. PMID- 8400838 TI - Drinking patterns and the gender mix of occupations: evidence from a national survey of American workers. AB - The linkage of the gender mix of occupations to drinking patterns has been suggested by Wilsnack and Wilsnack (1991). Using a national sample of American workers, associations among gender, the gender mix of occupations, occupation, and drinking variables were explored. The results suggest that the relationship between the gender mix of occupations and drinking variables operates through opportunities to drink with coworkers. Gender mix is associated with opportunities to drink with coworkers. Opportunities to drink with coworkers are, in turn, associated with whether respondents drink, who they are with when they drink, average number of drinks per month, and CAGE scores. Gender is associated with opportunities to drink with coworkers, drinks per month, and CAGE scores. Occupation is associated with opportunities to drink with coworkers, drinking patterns, and problem drinking. Further elaboration of the mechanisms linking the gender mix of occupations and drinking patterns is warranted. PMID- 8400839 TI - Drug use and cigarette smoking in applicants for drug abuse treatment. AB - Whereas smoking rates have declined in the last three decades from around 40% to under 30%, the rates for substance abuse clients remain at a plateau almost three times as high. In order to examine the relations between nicotine and drug use, applicants for drug abuse treatment were surveyed about their cigarette smoking. Results indicated that 85% of applicants were smokers. Also, it was found that for smokers, frequency and type of drug use were associated with amount of cigarette use. Among the five categories of primary drugs surveyed, heroin users had the highest levels of smoking. Additionally, frequency of primary drug use and frequency of overall drug use correlated with cigarette smoking. Within categories of primary drug, frequency of use was related to severity of nicotine dependence for marijuana only. Given the possible association between severity of illicit drug use and smoking found in this study, it behooves treatment organizations to help clients overcome their nicotine dependence. PMID- 8400840 TI - Pneumothorax in polysubstance-abusing marijuana and tobacco smokers: three cases. AB - Three patients are reported who suffered spontaneous pneumothorax, each of whom also had a history of daily marijuana and tobacco use. The patients ranged in age from 24 to 37 years, had smoked marijuana on a daily basis for 10 to 14 years, and had 11 to 50 pack-year tobacco-smoking histories. Marijuana may predispose to pneumothorax both by accelerating tobacco-induced lung disease, and by the frequent performance of the Valsalva maneuver during marijuana smoking. Although there is an association between pneumothorax and heavy tobacco use, an association with chronic daily marijuana use has not previously been reported. PMID- 8400841 TI - Young adult offspring and their families of origin: cohesion, adaptability, and addiction. AB - This study examined relationships among measures of family functioning (FACES II), number of family addictions, and risk of alcoholism (Short Michigan Alcoholism Screening Test, SMAST) in a college sample. Multiple regression analyses indicated that the number of family addictions and the additively combined FACES-II scales each predicted scores on the SMAST. Thus, addictions in the family contribute to offspring alcoholism both directly, through their presence, and indirectly, through family functioning. Information on the history of a wide range of family addictions and on the functioning style of the family of origin can be valuable in assessing and addressing addiction risk. PMID- 8400842 TI - Factors that influence the distribution of trace element-containing chemical species in living systems before and after sample collection. AB - The identification and characterisation of the various species of a trace element in a system could provide important clues as to its biochemical role(s). In order that the results of experiments reflect the true situation in the system under study at the time the sample was taken, chemical and physical changes that can affect the distribution profile of a given element have to be understood and controlled. A number of factors that can influence the nature and form of chemical species before, during and after sampling of biological systems have been identified and discussed. Precautions that could help in limiting such changes are recommended. PMID- 8400843 TI - Zinc deficiency and dietary folate metabolism in pregnant rats. AB - Five groups of pregnant Wistar rats (zinc-deficient diet without folate supplementation; folinic acid, folate monoglutamate, folate polyglutamate supplemented groups receiving zinc-deficient diet; pair-fed groups as controls) were fed from day one of fertilization with a semisynthetic zinc-deficient diet containing 0.2 mg/kg of Zn in the diet for the 4 deficient groups and 100 mg/kg for the pair-fed group. After 20 days, the zinc status (plasma, liver, femoral bone) was significantly decreased in the zinc-deficient groups. The liver and plasma folate levels were lower in the zinc-deficient groups compared to the pair fed group. Moreover, the folinic acid and the polyglutamate folate supplementations (100 mg/kg diet) did not normalize the folate status of the animals. Only the supplementation with folate monoglutamate led to correct folate levels in the pregnant rats. Nevertheless, no form of folate supplementation prevented fetal growth retardation in any of the zinc-deficient groups. These results indicate that zinc deficiency in pregnant rats decreases folate bioavailability of folinic acid, folate polyglutamates and, to a lesser extent, that of folate monoglutamate. However, no form of folate supplementation (i.e., folate monoglutamate) prevents fetal growth defect and the incidence of malformation in zinc-deficient rats. PMID- 8400844 TI - Changes in serum thyroid hormone levels and urinary ketone body excretion caused by a low selenium diet or silver loading in rats. AB - Biochemical indices of selenium (Se) deficiency (tissue Se content and glutathione peroxidase activity, serum thyroid hormone level, and urinary ketone bodies during fasting) were measured in rats fed a low-Se diet or rats loaded with silver (Ag). One group of male weanling Wistar rats was fed a casein-based low-Se basal diet (Se content, 0.027 mg/kg). Other groups were fed the basal diet supplemented with sodium selenite (0.2 mg/kg of Se), or silver acetate (250 mg/kg of Ag), or both for 6 weeks. Without silver loading, hepatic and blood Se contents and GSH-Px activities were much lower in the rats fed the basal diet than in the rats fed the selenite-supplemented diet. The Ag loading decreased the Se contents and GSH-Px activities irrespective of dietary Se level. There were significantly higher serum thyroxine (T4) concentrations in rats fed the low-Se diet and in Ag-loaded rats than in rats fed the selenite-supplemented diet without added Ag. Differences in serum 3'-, 5, 3-triiodothyronine (T3) concentration were not significant among the 4 groups. Urinary ketone body excretion was abnormally high in rats fed the basal low-Se diet, but Ag-loading decreased the ketone body excretion; ketone body metabolism in the Ag-induced low Se state was different from that in the diet-induced low-Se state. PMID- 8400845 TI - Bioavailability of zinc in rats fed on tuna as a protein source in the diet. AB - The objective of this study was to investigate the dietary zinc bioavailability in diets containing raw, cooked, steamed, and canned tuna as a protein source in the diet, and to evaluate different means of preparation, as well as various storage periods of the canned tuna, which was prepared according to industrial standards. Sterilized oil-free canned tuna was also made. The biological assays used for the study of the zinc utilization were carried out on Wistar rats fed with semi-synthetic diets for 12 days varying only in the protein source, which was casein-methionine or tuna provided as follows: raw, cooked in brine, steamed, sterilized with or without soybean oil, canned and stored for one or three years. Body weight, food intake, faecal and urinary zinc excretion, and zinc content in rat liver were monitored. No modifications were shown in food efficiency when diets including tuna were used, compared with those values from the casein methionine diet. Thus, final body weight results were similar in all groups tested. Dietary zinc absorption was enhanced by consuming a raw white tuna diet. This beneficial effect disappeared after steaming or cooking in brine, but it was slightly recovered after sterilization with oil and storage of the canned tuna. Animals fed on the diets including the oil-free canned tuna, showed two or three times higher faecal and urinary zinc excretion than those fed on casein and canned tuna stored for 0, 1 or 3 years, reaching a negative balance. A negative effect was therefore found in both dietary and endogenous zinc utilization due to the consumption of oil-free canned tuna. PMID- 8400846 TI - Concentration of free calcium in erythrocytes of lead-depleted rats. AB - The concentration of Ca(2+)-ions in erythrocytes of lead-depleted rats was estimated using the fluorescent Ca(2+)-indicator fura-2. The lead-depleted animals of the F1 generation showed an elevated Ca(2+)-concentration, whereas in the Po generation there was no difference in the Ca(2+)-concentration between depletion and control group. The elevation of Ca(2+)-ion concentration of lead depleted rats could be caused by a reduction in Ca-Mg-ATPase activity in lead depletion and could possibly lead to a decreased mean corpuscular volume (MCV) of erythrocytes. PMID- 8400847 TI - Zinc absorption in experimental osmotic diarrhea: effect of long-chain fatty acids. AB - The effect of free fatty acids on zinc absorption was studied in a rat model of chronic osmotic diarrhea induced with magnesium citrate and phenolphthalein. In vivo rates of zinc removal from the lumen and analysis of tissue for zinc uptake and metallothionein alterations were monitored. One mmol/L stearate enhanced zinc absorption in rats with or without diarrhea, from 207 +/- 22 and 353 +/- 13 pmol/min x cm to 676 +/- 34 and 610 +/- 26 pmol/min x cm, respectively. Palmitate was only effective in normal rats. Zinc absorption inversely correlated with mucosal zinc content in the perfused intestinal segments, in both type of rats. Hepatic metallothionein was enhanced by zinc and even more by oleate plus zinc in both groups; kidney metallothionein in animals with diarrhea was normalized by either oleate or zinc. The data support previous reports on the effect of long chain fatty acids on the enhancement of zinc absorption: saturation and a longer chain appear to be positive factors. A membrane modification role of long-chain fatty acids could have nutritional implications in the formulation of special diets. PMID- 8400848 TI - Perinatal development of iron and antioxidant defence systems. AB - The time course of malondialdehyde (MDA), Fe, vitamin E, superoxide dismutase (Cu, Zn-SOD) and glutathione peroxidase (GSH-Px) was measured in serum, liver, kidney and heart from rats between day 17 of gestation and day 20 after birth. MDA was higher in fetal than maternal tissues and showed a transient increase after birth. The low maternal MDA levels were reached later than 3 weeks after birth. Fe was higher in fetal than in maternal serum and liver. Five days after birth Fe drastically dropped in these tissues. The antioxidant factors vitamin E, Cu, Zn-SOD and GSH-Px were low in fetal tissues and rose after birth. It is discussed whether lipid peroxidation is increased in fetal tissues because of less-developed antioxidant defence mechanisms. PMID- 8400850 TI - Review of publications. PMID- 8400849 TI - Increased plasma T4-levels in children with low selenium state due to reduced type I iodothyronine 5'deiodinase activity? PMID- 8400851 TI - Application of flow injection analysis in trace element determination of body fluids. AB - The principle of flow injections analysis (FIA) is briefly described, along with some of its advantages for analytical measurements. A review of the development, current status and application of FIA coupled with optical or electrochemical techniques for the determination of various elements in body fluids is presented. Further, the analytical potential of FIA for the determination of elements in biological materials is then summarized and trends in the methodology are given brief consideration. PMID- 8400852 TI - Recent developments in melanoma epidemiology, 1993. AB - Recent work on melanoma epidemiology up to and including the Third International Conference in 1993 shows that in several countries mortality trends have stabilized or are decreasing, although incidence continues to increase. Despite specific attention, evidence for a role of factors other than sun and ultraviolet exposure is weak. The established aetiological factors are sun exposure, particularly intermittent exposure, and exposure to artificial sources of ultraviolet radiation. Acquired naevi are also related to sun exposure and may be a useful biological marker, as may be mutations such as p53. Evidence for the effectiveness of educational efforts to reduce sun exposure of populations is now provided by Australian work, and studies in Scotland demonstrate the effectiveness of public education in early diagnosis. A major unresolved issue is the value or otherwise of population screening, and systematic trails are required. PMID- 8400853 TI - The melanocyte-stimulating hormone (MSH) receptor in M2R mouse melanoma tumours: solubilization and properties of the receptor-MSH complex and its covalently crosslinked conjugate. AB - Several properties of the MSH receptor in solid melanotic and amelanotic mouse M2R tumour isografts were studied in C57BL mice. Using cell membrane fractions prepared from such tumours and the superpotent [Nle4,D-Phe7]alpha MSH analogue, the affinity and receptor contents of the two tumour variants were found to be similar. When occupied by MSH, the receptor-MSH complex (R.MSH) was readily soluble in cholate. In the solubilized form, R.MSH was extremely stable and dissociated to an extent of only 30% within 12 days at 4 degrees C. While this high stability can be maintained in the pH range of 7.0-8.5, the solubilized R.MSH complex becomes increasingly unstable below pH 7.0 and totally dissociates at a pH < 6.0. In the membrane-bound form, the R.MSH complex shows a parallel pH stability profile which is shifted down by approximately two pH units. In addition to low pH, the R.MSH complex becomes unstable and totally dissociates in the presence of 10 mM EGTA, suggesting that the calcium-sensitive function of the receptor is still associated with the receptor in the detergent-soluble state. The R.MSH complexes in the soluble and membrane-bound forms are also totally resistant to proteolytic digestion by V8 protease, but were slowly digested by trypsin. Treatment of R.MSH with 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide hydrochloride or bis (sulphosuccinimidyl) suberate led to covalent crosslinking of MSH to the receptor molecule. The electrophoretic mobility on SDS-PAGE of the 43/46 kD doublet of the receptor-MSH conjugate (R*MSH) was identical to the photoaffinity labelled MSH receptor product described earlier in cultured M2R cells. However, the efficiency of production of the crosslinked product was approximately 30%, much higher than that achieved previously by photoaffinity labelling. Using rabbit polyclonal anti-alpha MSH antibodies, the R*MSH conjugate was identifiable on Western immunoblots. These results provide a basis for further development of procedures for purification of the MSH receptor molecule and studying its protein structure. PMID- 8400854 TI - Cytogenetic findings in 20 melanomas. AB - We report cytogenetic studies performed on 20 patients with cutaneous malignant melanoma, characterized by clinical and histological parameters. Cytogenetic analyses were performed on peripheral blood lymphocytes, in order to exclude the presence of constitutional chromosomal aberrations, and on primary cell cultures obtained from neoplastic skin lesions. A metastasis was also cultured in order to characterize chromosome markers. Specific markers found in more than one patient were t(1;14)(q21;q32) and aberrations of the 4q21,8q24 and 10q24q26 regions. The research aims to identify possible subtypes of melanomas related to specific chromosomal markers. It is hoped that this will contribute to understanding of the aetiology and evolution of the disease in order to obtain a more exact classification. We compare our results with the data reported in the literature and discuss the possible role of the cytogenetic analyses in human malignant melanoma. PMID- 8400855 TI - Properties of melanosomes and their exploitation in the diagnosis and treatment of melanoma. AB - Melanosomes are subcellular organelles with unique properties: they are producers and scavengers of cytotoxic quinones and are involved in free radical reactions. They behave as energy absorbing and converting devices, have an ion-exchange capacity and show affinity to a variety of substances. Even in the absence of melanin, premelanosomes show characteristic properties due to the presence of specific proteins. The melanosome concentration in pigment tissues is high enough to offer possibilities for influencing pigment cells and for detecting and/or killing melanoma cells. The exploitation of melanosome properties in practice is reviewed. PMID- 8400856 TI - Ocular melanoma: an experimental model in New Zealand white rabbits. AB - An experimental model is suggested for reproducing ocular melanoma in New Zealand white rabbits using B16 melanoma cells and protocols differing with respect to either tumour origin (subcutaneous fragments of melanoma B16 or B16-F10 tumour cell cultures) or implant site (the anterior chamber or subchoroidal). In 20 animals, 20 mg of methylprednisolone acetate was injected subconjunctivally as a local immunosuppressant. The only protocol resulting in tumour was inoculation of 4 x 10(6) B16-F10 melanocytes into the anterior chamber of the eye. Trans-scleral injections of cell suspensions produced tumour growth in 43% (13/30) of animals so treated. Thirteen animals developed non-neoplastic pigmented lesions formed of numerous melanophages. Another 19 animals showed non-pigmented lesions caused by reaction to the surgical procedures. Subconjunctivally injected methylprednisolone acetate did not increase the incidence of tumour growth. PMID- 8400857 TI - Heterogeneity of the T-cell and macrophage infiltrate in cutaneous and subcutaneous metastases of melanoma patients. AB - To examine the intra-individual heterogeneity of the local inflammatory infiltrate in metastases of untreated melanoma patients we analysed 42 skin and subcutaneous metastases from 11 patients. Metastases were excised on the same date (10 patients) or consecutively (six patients), and processed in a two-step immunoperoxidase technique using monoclonal antibodies directed against T-cells (CD3, CD25) and two macrophage subpopulations (RM 3/1, 27E10). The number of positively stained T-cells and macrophages were counted for each lesion in representative high power fields. The mean values were calculated. We observed limited heterogeneity among T-cells (CD3+) and activated T-cells (CD25+) in simultaneously excised metastases. Contrary to this, markedly heterogeneous CD3+ infiltrates were found in metastases taken on different dates, even if there was a time difference of only 1 day between excisions, suggesting spontaneous variations. However, activated T-cells in follow-up biopsies showed only limited heterogeneity. In spite of inter-individual variations, metastases taken simultaneously, as well as consecutively, showed little intra-individual heterogeneity with respect to macrophages. Our findings should be taken into account for comparative studies of the infiltrate in metastatic melanoma during immunotherapy. PMID- 8400858 TI - Congenital myasthenic syndromes. AB - The Congenital Myasthenic Syndromes (CMS) constitute a group of rare genetic disorders affecting neuromuscular transmission. They differ from myasthenia gravis and the Lambert-Eaton myasthenic syndrome, which are autoimmune antibody mediated conditions. CMS can present at any time from birth to adulthood, though usually within the first 2 yr of life, and result in a spectrum of diseases ranging from mild weakness to severe disability with life-threatening episodes. Several of these syndromes have been well documented, and in recent years fully investigated using a variety of electrophysiological, histochemical, and morphological techniques. In this review we describe the main results of these investigations, and attempt to classify the disorders into groups that can be recognized by the clinician. They include defects in acetylcholine release, absence of the endplate-specific form of acetylcholinesterase, and alterations in the number or function of postsynaptic acetylcholine receptors. Clinical features are described in detail, and treatment reviewed. These disorders involve a potentially large number of candidate genes. Further elucidation of the underlying abnormalities will not only lead to improved treatment, but should contribute to our understanding of the molecular biology of the neuromuscular junction. PMID- 8400859 TI - MELAS point mutation with unusual clinical presentation. AB - Mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes (MELAS) is a multisystemic mitochondrial disorder (Pavlakis et al. Advances in Contemporary Neurology. Philadelphia: Davis, 1988: 95-133) and most patients with the typical MELAS phenotype have a point mutation in mitochondrial DNA, an A to G transition at nucleotide 3243 (Goto et al. Nature 1990; 348; 651-653; Koboyashi et al. Biochem Biophys Res Commun 1990; 173: 816-822; Ciafaloni et al. Ann Neurol 1992; 31: 391-398). A 9-yr-old boy presenting with chronic asthma and depression was found to have abnormal mitochondria, partial defects of respiratory chain enzymes, and the MELAS point mutation. PMID- 8400860 TI - Androgen receptor gene polymorphisms in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is more common in men than in women (male to female ratio of approximately 2:1), suggesting a role for a sex-linked factor in the disease. The recent identification of a mutation of the androgen receptor gene in Kennedy's disease or X-linked bulbospinal neuronopathy, a rare form of progressive lower motor neurone degeneration, also associated with clinical signs of androgen insensitivity, raises the possibility that androgen function may be disturbed in other motor neurone disorders, including ALS. The Kennedy's disease mutation consists of an increased size of a highly polymorphic CAG repeat sequence in the first exon of the androgen receptor gene, coding for a polyglutamine tract. We have analysed this CAG repeat sequence in a large number of patients with typical sporadic ALS and in normal controls, in order to test the hypothesis that this polymorphism of the androgen receptor gene may influence susceptibility for ALS. We report that the distribution of alleles relating to the size of the CAG repeat sequence of the androgen receptor gene is similar in ALS and controls, indicating that polymorphisms of the CAG repeat sequence of the androgen receptor gene play a limited role, if any, in susceptibility to ALS. PMID- 8400861 TI - Spine surgery in spinal muscular atrophy: long-term results. AB - Fifteen patients with spinal muscular atrophy operated on for scoliosis were reviewed and interviewed. Age at time of surgery averaged 16 yr. Follow-up averaged more than 5 yr. Eleven patients underwent posterior spinal fusion with Luque instrumentation. Four had Harrington instrumentation, with segmental wiring in three. Three out of the 15 also had anterior fusion with Dwyer instrumentation. Patients were immobilized in cast or brace for 6 months, on average, after surgery. The average curve correction was 50%. There was a loss of vital capacity after the operation, which in most cases was regained with time. In functional terms, there were improvements and deteriorations both in the short and long term. Esthetic appearance improved in all patients, as did the perceived well-being. Retrospectively all but one would have the operation again. PMID- 8400862 TI - Congenital sensory neuropathy in association with ichthyosis and anterior chamber cleavage syndrome. AB - Two patients with a congenital neuropathy are described. Both had atypical features including: ichthyosis and a mild anterior chamber cleavage syndrome. Both had severely reduced, or absent, sensation for light touch, vibration, position and temperature. Pain sensation was mildly reduced. There was some evidence of motor involvement but this was relatively minor compared with the sensory involvement. Nerve action potentials were small or absent and sural nerve biopsies showed almost complete absence of myelinated nerve fibres with multiple bundles of abnormally arranged axons and Schwann cell processes. These patients appear to have an undescribed syndrome in which the large sensory neurons and the anterior chamber of the eye did not develop properly. This may reflect a failure of migration, differentiation or proliferation of neural crest cells. PMID- 8400863 TI - Oral phosphate supplements reverse skeletal muscle abnormalities in a case of chronic fatigue with idiopathic renal hypophosphatemia. AB - A 57-yr-old man presented with a long history of undiagnosed fatigue but no evidence of bone disease. He was noted to have hypophosphatemia due to an idiopathic phosphaturia. Marked abnormalities of exercising skeletal muscle detected by phosphorus magnetic resonance spectroscopy and by plasma metabolite measurements were consistent with mitochondrial dysfunction. Oral phosphate supplements restored plasma phosphate concentration and muscle biochemistry to normal and produced considerable improvement in symptoms and exercise tolerance, although the phosphate concentration in muscle was only marginally low and increased little by treatment. We conclude that hypophosphatemia should be excluded in unexplained fatigue. PMID- 8400864 TI - Carcinomatous neuropathy associated with hepatic cell carcinoma: an autopsy case report. AB - A 76-yr-old male patient with carcinomatous neuropathy associated with hepatic cell carcinoma, whose initial symptom was deep sensory disturbance followed by muscle weakness is described. The onset was subacute, followed by slow progression. Sural nerve biopsy, as well as electrophysiological examinations, revealed severe axonal degeneration without any evidence of demyelination. The autopsy findings were similar to findings described in the literature on carcinomatous neuropathy. Although carcinomatous neuropathy is usually associated with lung cancer, this report describes an association with hepatic cell carcinoma. The patient also had motor nerve involvement with positive serum anti GM1 ganglioside antibody which decreased after immunosuppressant therapy in parallel with recovery of muscle weakness. The anti-GM1 ganglioside antibody may be involved in the pathogenesis of motor disturbance in the present case. PMID- 8400865 TI - Neonatal screening for muscular dystrophy. Consensus recommendation of the 14th workshop sponsored by the European Neuromuscular Center (ENMC). PMID- 8400866 TI - Report on the 16th ENMC workshop--carrier diagnosis of Duchenne and Becker muscular dystrophy. PMID- 8400867 TI - Neuromuscular disorders: gene location. PMID- 8400868 TI - Distinct cis-acting elements direct pistil-specific and pollen-specific activity of the Brassica S locus glycoprotein gene promoter. AB - The promoter of the S Locus Glycoprotein (SLG) gene of Brassica is a tightly regulated promoter that is active specifically in reproductive organs. In transgenic tobacco, this promoter is active exclusively in cells of the pistil and in pollen. We transformed tobacco with truncated versions of the SLG13 promoter fused to the beta-glucuronidase reporter gene. We show that the promoter has a modular organization and consists of separable DNA elements that independently specify pistil- and pollen-specific expression. A 196-bp region ( 339 to -143) is sufficient to confer stigma and style specificity to the marker gene. Two distinct, but functionally redundant, domains (-415 to -291 and -117 to -8) allow specific expression of the gene in pollen. The functional domains identified within the SLG13 promoter contain sequence elements that are highly conserved in different alleles of the SLG gene and in the S Locus Related SLR1 gene. PMID- 8400869 TI - RPS2, an Arabidopsis disease resistance locus specifying recognition of Pseudomonas syringae strains expressing the avirulence gene avrRpt2. AB - A molecular genetic approach was used to identify and characterize plant genes that control bacterial disease resistance in Arabidopsis. A screen for mutants with altered resistance to the bacterial pathogen Pseudomonas syringae pv. tomato (Pst) expressing the avirulence gene avrRpt2 resulted in the isolation of four susceptible rps (resistance to P. syringae) mutants. The rps mutants lost resistance specifically to bacterial strains expressing avrRpt2 as they retained resistance to Pst strains expressing the avirulence genes avrB or avrRpm1. Genetic analysis indicated that in each of the four rps mutants, susceptibility was due to a single mutation mapping to the same locus on chromosome 4. Identification of a resistance locus with specificity for a single bacterial avirulence gene suggests that this locus, designated RPS2, controls specific recognition of bacteria expressing the avirulence gene avrRpt2. Ecotype Wu-0, a naturally occurring line that is susceptible to Pst strains expressing avrRpt2, appears to lack a functional allele at RPS2, demonstrating that there is natural variation at the RPS2 locus among wild populations of Arabidopsis. PMID- 8400870 TI - A 22-bp fragment of the pea lectin promoter containing essential TGAC-like motifs confers seed-specific gene expression. AB - To elucidate the molecular mechanisms responsible for seed-specific gene expression in plants, the promoter of the pea lectin (psl) gene, encoding an abundant seed protein, was used as a model. Leaf and seed nuclear proteins bound to a region in the psl promoter containing three overlapping TGAC-like motifs, which have been shown to be a binding site for basic/leucine zipper proteins, including TGA1a. A trimer of a 22-bp region of the psl promoter, containing the TGAC-like motifs, coupled to a heterologous minimal promoter conferred low reporter gene expression in root, stem, and leaf and high expression in seed of transgenic tobacco. Expression increased during the midmaturation stage of seed development and was observed in the endosperm as well as in the embryo, where it strongly decreased within a few days after germination. This expression pattern is qualitatively identical to the expression pattern conferred by a 2000-bp fragment of the psl promoter. Nucleotides within the TGAC-like motifs important for in vitro binding are also essential for in vivo transcription activation in vegetative tissue as well as in seed. The electrophoretic mobility of a DNA protein complex containing seed nuclear protein was different from that formed with leaf nuclear protein. Furthermore, the TGA1a steady state mRNA level in immature seed was relatively low. These results suggest that a seed-specific factor different from TGA1a, but with similar binding specificity, is responsible for gene activation in seed. We conclude that the 22-bp region contains all the information, including an essential TGAGTCATCA sequence, necessary for seed specific expression and very likely plays an essential role in the seed-specific expression pattern of the psl gene. PMID- 8400871 TI - Mutations at the SPINDLY locus of Arabidopsis alter gibberellin signal transduction. AB - Three independent recessive mutations at the SPINDLY (SPY) locus of Arabidopsis confer resistance to the gibberellin (GA) biosynthesis inhibitor paclobutrazol. Relative to wild type, spy mutants exhibit longer hypocotyls, leaves that are a lighter green color, increased stem elongation, early flowering, parthenocarpy, and partial male sterility. All of these phenotypes are also observed when wild type Arabidopsis plants are repeatedly treated with gibberellin A3 (GA3). The spy 1 allele is partially epistatic to the ga1-2 mutation, which causes GA deficiency. In addition, the spy-1 mutation can simultaneously suppress the effects of the ga1-2 mutation and paclobutrazol treatment, which inhibit different steps in the GA biosynthesis pathway. This observation suggests that spy-1 activates a basal level of GA signal transduction that is independent of GA. Furthermore, results from GA3 dose-response experiments suggest that GA3 and spy-1 interact in an additive manner. These results are consistent with models in which the SPY gene product regulates a portion of the GA signal transduction pathway. PMID- 8400872 TI - Targeting of glyoxysomal proteins to peroxisomes in leaves and roots of a higher plant. AB - Higher plants possess several classes of peroxisomes that are present at distinct developmental stages and serve different metabolic roles. To investigate the cellular processes that regulate developmental transitions of peroxisomal function, we analyzed the targeting of glyoxysomal proteins to leaf-type and root peroxisomes. We transferred genes encoding the glyoxysome-specific enzymes isocitrate lyase (IL) and malate synthase into Arabidopsis plants and showed, in cell fractionation and immunogold localization experiments, that the glyoxysomal proteins were imported into leaf-type and root peroxisomes. We next defined the sequences that target IL to peroxisomes and asked whether the same targeting determinant is recognized by different classes of the organelle. By localizing deletion and fusion derivatives of IL, we showed that the polypeptide's carboxyl terminus is both necessary for its transport to peroxisomes and sufficient to redirect a passenger protein from the cytosol to both glyoxysomes and leaf-type peroxisomes. Thus, glyoxysomal proteins are transported into several classes of peroxisomes using a common targeting determinant, suggesting that protein import does not play a regulatory role in determining a peroxisome's function. Rather, the specific metabolic role of a peroxisome appears to be determined primarily by processes that regulate the synthesis and/or stability of its constituent proteins. These processes are specified by the differentiated state of the cells in which the organelles are found. PMID- 8400873 TI - Alanine scanning mutagenesis of a plant virus movement protein identifies three functional domains. AB - Alanine scanning mutagenesis was performed on the red clover necrotic mosaic virus (RCNMV) movement protein (MP), and 12 mutants were assayed in vitro for RNA binding characteristics and in vivo for their ability to potentiate RCNMV cell-to cell movement. The mutant phenotypes that were identified in vitro and in vivo suggest both that cooperative RNA binding is not necessary for cell-to-cell movement in vivo and that only a fraction of the wild-type RNA binding may be required. The MP mutants defined at least three distinct functional regions in the MP: an RNA binding domain, a cooperative RNA binding domain, and a third domain that is necessary for cell-to-cell movement in vivo. This third domain may be required for targeting the MP to cell walls and plasmodesmata, interacting with host proteins, folding, or possibly binding RNA into a functional ribonucleoprotein complex capable of cell-to-cell movement. PMID- 8400874 TI - Specific binding of nuclear localization sequences to plant nuclei. AB - We have begun to dissect the import apparatus of higher plants by examining the specific association of nuclear localization sequences (NLSs) with purified plant nuclei. Peptides to the simian virus 40 (SV40) large T antigen NLS and a bipartite NLS of maize were allowed to associate with tobacco and maize nuclei. Wild-type NLSs were found to compete for a single class of low-affinity binding sites having a dissociation constant (Kd) of approximately 200 microM. Peptides to mutant NLSs, which are inefficient in stimulating import, were poor competitors, as were reverse wild-type and non-NLS peptides. The NLS binding site was proteinaceous and resistant to extraction under conditions where pores were still associated. In addition, immunofluorescence and immunoelectron microscopy indicated that binding was at the nuclear envelope. Overall, plant nuclei may be an excellent system to identify components of the import apparatus. PMID- 8400875 TI - Suppressors of trp1 fluorescence identify a new arabidopsis gene, TRP4, encoding the anthranilate synthase beta subunit. AB - Suppressors of the blue fluorescence phenotype of the Arabidopsis trp1-100 mutant can be used to identify mutations in genes involved in plant tryptophan biosynthesis. Two recessive suppressor mutations define a new gene, TRP4. The trp4 mutant and the trp1-100 mutant are morphologically normal and grow without tryptophan, whereas the trp4; trp1-100 double mutant requires tryptophan for growth. The trp4; trp1-100 double mutant does not segregate at expected frequencies in genetic crosses because of a female-specific defect in transmission of the double mutant genotype, suggesting a role for the tryptophan pathway in female gametophyte development. Genetic and biochemical evidence shows that trp4 mutants are defective in a gene encoding the beta subunit of anthranilate synthase (AS). Arabidopsis AS beta subunit genes were isolated by complementation of an Escherichia coli anthranilate synthase mutation. The trp4 mutation cosegregates with one of the genes, ASB1, located on chromosome 1. Sequence analysis of the ASB1 gene from trp4-1 and trp4-2 plants revealed different single base pair substitutions relative to the wild type. Anthranilate synthase alpha and beta subunit genes are regulated coordinately in response to bacterial pathogen infiltration. PMID- 8400876 TI - Positive and negative regulatory regions control the spatial distribution of polygalacturonase transcription in tomato fruit pericarp. AB - The tomato fruit consists of a thick, fleshy pericarp composed predominantly of highly vacuolated parenchymatous cells, which surrounds the seeds. During ripening, the activation of gene expression results in dramatic biochemical and physiological changes in the pericarp. The polygalacturonase (PG) gene, unlike many fruit ripening-induced genes, is not activated by the increase in ethylene hormone concentration associated with the onset of ripening. To investigate ethylene concentration-independent gene transcription in ripe tomato fruit, we analyzed the expression of chimeric PG promoter-beta-glucuronidase (GUS) reporter gene fusions in transgenic tomato plants. We determined that a 1.4-kb PG promoter directs ripening-regulated transcription in outer pericarp but not in inner pericarp cells, with a sharp boundary of PG promoter activity located midway through the pericarp. Promoter deletion analysis indicated that a minimum of three promoter regions influence the spatial regulation of PG transcription. A positive regulatory region from -231 to -134 promotes gene transcription in the outer pericarp of ripe fruit. A second positive regulatory region from -806 to 443 extends gene activity to the inner pericarp. However, a negative regulatory region from -1411 to -1150 inhibits gene transcription in the inner pericarp. DNase I footprint analysis showed that nuclear proteins in unripe and ripe fruit interact with DNA sequences within each of these three regulatory regions. Thus, temporal and spatial control of PG transcription is mediated by the interaction of negative and positive regulatory promoter elements, resulting in gene activity in the outer pericarp but not the inner pericarp of ripe tomato fruit. The expression pattern of PG suggests that, although they are morphologically similar, there is a fundamental difference between the parenchymatous cells within the inner and outer pericarp. PMID- 8400877 TI - Arabidopsis HY8 locus encodes phytochrome A. AB - hy8 long hypocotyl mutants of Arabidopsis defective in responsiveness to prolonged far-red light (the so-called "far-red high-irradiance response") are selectively deficient in functional phytochrome A. To define the molecular lesion in these mutants, we sequenced the phytochrome A gene (phyA) in lines carrying one or other of two classes of hy8 alleles. The hy8-1 and hy8-2 mutants that express no detectable phytochrome A each have a single nucleotide change that inserts a translational stop codon in the protein coding sequence. These results establish that phyA resides at the HY8 locus. The hy8-3 mutant that expresses wild-type levels of photochemically active phytochrome A has a glycine-to glutamate missense mutation at residue 727 in the C-terminal domain of the phyA sequence. Quantitative fluence rate response analysis showed that the mutant phytochrome A molecule produced by hy8-3 exhibited no detectable regulatory activity above that of the phyA-protein-deficient hy8-2 mutant. This result indicates that glycine-727, which is invariant in all sequenced phytochromes, has a function important to the regulatory activity of phytochrome A but not to photoperception. PMID- 8400879 TI - The proposed plant connexin is a protein kinase-like protein. PMID- 8400878 TI - Jordan, an active Volvox transposable element similar to higher plant transposons. AB - We have isolated a 1595-bp transposable element from the multicellular green alga Volvox carteri following its insertion into the nitrate reductase (nitA) locus. This element, which we have named Jordan, has short (12-bp) terminal inverted repeats and creates a 3-bp target site duplication, like some higher plant transposons of the classic type. Contained within the first 200 bp of one end of the element are 55-bp inverted repeats, one of which begins with the terminal inverted repeat. Revertants of the transposon insertion into the nitA locus were obtained at a rate of approximately 10(-4) per Volvox embryo per generation. In each revertant examined, all transposon sequences were completely excised, but footprints containing both sets of duplicated bases, in addition to three to nine extra bases, were left behind. Jordan contains no significant open reading frames and so appears to be nonautonomous. DNA gel blot analysis indicates that Jordan is a member of a large family of homologous elements in the Volvox genome. We have isolated and characterized several of these homologs and found that they contain terminal very similar to those of Jordan. Efforts to utilize Jordan and its homologs as tools to tag and clone developmentally interesting genes of Volvox are discussed. PMID- 8400880 TI - Human gamma delta T lymphocytes. PMID- 8400881 TI - Suppressive effect of 2-acetyl-4-tetrahydroxybutylimidazole on contact hypersensitivity in the Skh: HR hairless mouse. AB - The compound 2-acetyl-4-tetrahydroxybutylimidazole (THI), a component of ammonia caramel, has been shown to cause lymphopenia and to impair several immune functions in rats and mice. In this study we show that THI effectively suppresses contact hypersensitivity dose responsively in the hairless mouse, whether administered topically or orally. The suppression was shown to be prevented by topical administration of the histamine antagonist, cimetidine, and by the dipeptide, carnosine. Splenocytes from THI-treated mice failed to elicit normal contact hypersensitivity when transferred to naive mice. This suggests that THI acts by modifying splenocyte function, perhaps via a histamine-like receptor site(s). PMID- 8400882 TI - Recurrent angioedema caused by circulating immune complexes containing antibodies against bovine proteins. AB - A 27-year-old woman with severe recurrent angioedema and urticaria since 5 years was found to have high levels of circulating immune complexes in the peripheral blood. These immune complexes contained antibodies against bovine serum albumin. Elimination of bovine products from the diet resulted in disappearance of immune complexes within 2 days and addition again to the diet led to reappearance within 24 h. The patient has now been in complete remission during 4.5 years under treatment with a diet free of bovine proteins but containing products from chicken, egg, pork and fish. The immune complexes have not reappeared. Thus, immune complexes containing antibodies against alimentary antigens can provoke acute vascular damage manifested as angioedema and urticaria. In such cases, elimination of the causative antigen from the diet can result in patient being cured. PMID- 8400883 TI - Activation of multiple protein kinases including a MAP kinase upon Fc epsilon RI cross-linking. AB - Previous studies have shown that protein-serine/threonine kinases and protein tyrosine kinase(s) are activated by cross-linking of the high-affinity receptor for IgE, Fc epsilon RI, on mast cells and basophils. In vitro kinase assays (ISDR kinase assays) on cellular proteins immobilized on polyvinylidene difluoride membrane after denaturation and renaturation were employed to estimate the complexity of protein kinases expressed in mouse mast cells. The results demonstrated that a large number (more than 60) of both serine/threonine- and tyrosine-specific kinases are present in a mouse mast cell line, PT-18. Cross linking of Fc epsilon RI-induced activation of a subset of both serine/threonine kinases and tyrosine kinases in PT-18 as well as bone marrow-derived mouse mast cells, as revealed by the ISDR kinase assay. Among them, MAP kinase (or ERK2) was shown to be tyrosine phosphorylated and activated transiently upon Fc epsilon RI cross-linking, suggesting its potential role in mast cell signal transduction. PMID- 8400884 TI - Ultrastructural identification of exocytosis of granules from human gut eosinophils in vivo. AB - Twenty-two percent of 117 biopsies of human intestinal tissues had ultrastructural images of classical regulated secretion from eosinophils in vivo i.e. eosinophil granule extrusion (EGE). Replicate intestinal biopsies that were positive for bacteria had EGE more often than not (p < 0.05); 77% of the isolates were Staphylococci. Some of the intestinal biopsies also had damaged nerves; all that had EGE and damaged enteric nerves also had positive bacterial cultures. The EGE that we observed could not account for all enteric nerve damage, suggesting multifactorial mechanisms for nerve damage in gut tissues. Among the possibilities are release of neurotoxic eosinophil granule proteins by an alternate secretory route, i.e., piecemeal degranulation, direct toxicity of tissue invasive bacteria and/or damaged nerves of unknown etiology such as those that are regularly present in uninvolved tissues of patients with Crohn's disease. PMID- 8400885 TI - Immunization of rabbits with spermine induces antibodies to self antigens. AB - Rabbits were immunized under different schedules with spermine in the free form or with random noncovalent complexes of spermine or spermidine with ovalbumin. The specificity of the induced antibodies was determined by ELISA and by dot immunobinding assay. Our results show that in vitro conjugation of spermine and spermidine to a carrier is not an obligatory prerequisite for obtaining corresponding antibodies. Anti-spermine antibodies were found in 9 of 19 animals injected with spermine. Furthermore, all 19 rabbits produced distinct populations of IgM and IgG antibodies which reacted with histones, various synthetic peptides of histones, as well with ubiquitin, a peptide of ubiquitin, dsDNA and two 29 base 5' synthetic oligodeoxynucleotides. Even in antisera with no detectable reactivity with spermine, antibodies to some of the unrelated antigens were found. The pattern of reactivity of the antisera with the various antigens was different for each immunized animal. These findings lend support to the view that polyamines may play a role in the appearance of autoantibodies. PMID- 8400886 TI - IgE reactivity to 14-kD and 27-kD natural rubber proteins in latex-allergic children with spina bifida and other congenital anomalies. AB - We examined by immunoblotting sera from 39 latex-allergic patients, 19 from USA and 20 from Finland, including 15 children with spina bifida and 5 children with other congenital anomalies, for IgE antibodies to natural rubber latex (latex) antigens. IgE antibodies in 10 of the 12 (83%) US spina bifida patients and in 2 of the 3 Finnish spina bifida patients recognized a previously undescribed 27-kD antigen which, together with a 14- and a 20-kD peptide, appeared to be a major latex allergen. Two patients with other congenital anomalies, one from the US and one from Finland, also demonstrated anti-27-kD bands. IgE antibodies to the 27-kD antigen were not detected in other latex-allergic patients from the US or Finland although most of them showed IgE binding to 14- and/or 20-kD latex antigens. All 21 controls, including 3 spina bifida patients with no evidence of latex allergy, gave negative immunoblot results. This observation suggests that patients with spina bifida or other congenital anomalies who have been subject to multiple operations and other invasive therapeutic procedures may have been exposed to different antigenic source materials than other latex-allergic patients. PMID- 8400887 TI - Monoclonal antibodies against farmer's lung antigens having specific binding to IgG antibodies. AB - Hypersensitivity pneumonitis resulting from environmental exposure to Saccharopolyspora rectivirgula (Micropolyspora faeni) among farmers has been well recognized. The diagnosis of the disease depends on demonstration of circulating antibodies against S. rectivirgula. However, dependable pure antigens are not available for serodiagnosis. In the present study we have employed hybridoma technology to obtain monoclonal antibodies against S. rectivirgula antigens. These monoclonal antibodies were employed to purify antigens through affinity chromatography. When tested in ELISA, high levels of antibodies were demonstrated against these antigens in farmer's lung patient sera compared to exposed but asymptomatic individuals from the same household. In Western blots patient sera reacted with components of crude antigens with molecular masses of 28, 35, 60, 65 and 68 kD and 4 components above 100 kD, while the monoclonal antibodies reacted only with the 60-kD protein. These purified antigens can be used as reliable reagents in the specific diagnosis of farmer's lung disease. PMID- 8400888 TI - Two-dimensional gel electrophoretic analysis with immobilized pH gradients of Dactylis glomerata pollen allergens. AB - We studied Orchard grass pollen allergens (Dactylis glomerata) by a new two dimensional electrophoretic technique which is the combination of isoelectric focusing (IEF) in immobilized pH gradient (IPG) and horizontal SDS-PAGE. The IPG Dalt (two-dimensional gel electrophoresis with immobilized pH gradients in the first dimension) electrophoresis was made with four different but complementary pH gradients: 3-10.5, 4-9, 4-7, 6.3-10.5. We also compared the behavior of allergens in native and denaturing conditions during IEF. This technique followed by electroblotting on nitrocellulose membrane and immunodetection with an allergic patient serum was shown to increase the number of IgE-binding components up to 100 compared to one-dimensional techniques. Major allergen groups could be identified. New allergens not yet described could be characterized by their isoelectric points and their molecular masses. This highly resolving and reproducible technique, should allow an easy allergen standardization. PMID- 8400889 TI - Effect of cefadroxil on antigen-induced bronchial hyperresponsiveness and eosinophil accumulation in lung from sensitized guinea pigs. AB - The effect of a semi-synthetic cephalosporin, Cefadroxil, on antigen-induced bronchial hyperresponsiveness and eosinophil accumulation in lungs from sensitized guinea pigs was investigated and compared to the effects of Cetirizine and Ketotifen. When aerosol-sensitized guinea pigs were pretreated 1 h before the antigen challenge with Cefadroxil (100 mg/kg i.p.) a partial but significant inhibition of the bronchial hyperresponsiveness to aerosolized acetylcholine chloride was observed. Furthermore, the treatment of guinea pigs (115 mg/kg, per os) 24 and 1 h before ovalbumin challenge also significantly reduced bronchial hyperresponsiveness. In contrast, no significant inhibition was noted when the guinea pigs were treated by a single dose of Cefadroxil (115 mg/kg per os) 1 h before challenge. Pretreatment of the guinea pigs with Cetirizine (1 mg/kg per os) or Ketotifen (0.1 mg/kg per os) completely inhibited the antigen-induced bronchial hyperresponsiveness. Cefadroxil (100 mg/kg i.p.) slightly inhibited the accumulation of eosinophils in the peribronchial area induced by antigen challenge. In contrast, no significant reduction was noted when the guinea pigs were treated per os with Cefadroxil (115 mg/kg), Cetirizine (1 or 10 mg/kg) or Ketotifen (0.1 mg/kg). These results show that Cefadroxil is effective in reducing antigen-induced bronchial hyperresponsiveness, an effect independent of a reduction in the pulmonary inflammation, namely eosinophil accumulation in lung. PMID- 8400890 TI - Down-regulation of murine contact sensitivity by hapten-amino acid derivatives. AB - The low responsiveness of contact sensitivity (CS) in sensitized and challenged mice was induced by using hapten-amino acid derivatives. Subcutaneous injection of trinitrophenyl (TNP)-lysine or oxazolone-glycine inhibited ear swelling responses in mice that received rechallenge. The inhibitory effect was long lasting and antigen-specific. TNP-lysine blocked the transfer of CS in vivo and the binding of immune lymph node cells to antigen in vitro. Thus, the suppression was mediated by inhibition of effector cells, but not associated with the generation of efferent limb-acting suppressor cells. The tool reported herein seems to be quite useful for controlling ongoing murine CS to simple chemicals. PMID- 8400891 TI - Immunotherapy of allergic diseases. AB - Immunotherapy (IT) of IgE-mediated diseases has been used since its first description in 1911. This therapy has been used in patients with inhalant allergy and insect venom sensitivity. Unfortunately, IT suffers from the crudeness of the allergy extracts and the inability to define allergenic epitopes. Moreover, this form of IT has been increasingly questioned because of improvements in pharmacotherapy and the risks of untoward reactions. These untoward reactions even include anaphylaxis and death. Despite this criticism, there are increasing data on the efficacy of immunotherapy and considerable research is underway to improve the risk-benefit ratio of treatment. Other approaches directed at inhibiting the sequential events of both the afferent and efferent limbs of allergic reactions are in progress. This involves specific alterations of target cells, i.e., mast cells and basophils. Moreover, the development of recombinant allergens and the precise definition of the binding site of IgE will further promote research and development of new means of IT. Gene targeting therapy may follow, directed at T cells specific for allergen epitope(s) or IgE binding sites. The future holds great promise and excitement. PMID- 8400892 TI - Presence of anti-insulin reaginic auto-antibodies of the IgG4 class in insulin dependent (type I) diabetic patients before insulin therapy. AB - The autoimmune aetiology of type I diabetes has been well documented. We studied whether anti-insulin anaphylactic antibodies were present on the membrane of basophils from type I diabetics by the toluidine blue method (detecting basophil activation after stimulation by insulin). We observed that basophils of recently diagnosed insulin-dependent diabetic patients (n = 13) were statistically more frequently activated by insulin than basophils from noninsulin-dependent diabetics (p < 0.002, n = 8) or non-diabetic subjects (p < 0.05, n = 9). Basophils from normal donors were passively sensitized with plasma from insulin dependent diabetics and could then be activated by insulin. This sensitization still occurred when using plasma previously heated to 56 degrees C, indicating that the sensitizing antibodies were not of the IgE class. When basophils from type I diabetics were preincubated with anti-IgG subclasses, only anti-IgG4 monoclonal antibodies inhibited the insulin-induced basophil activation. By contrast, preincubation with blocking concentrations of anti-IgG1-3 antibodies or desensitization of the IgE pathway did not modify basophil activation. These experiments strongly suggest the presence of anti-insulin antibodies of the IgG4 subclass in insulin-dependent diabetics before any insulin administration and provide a simple tool to complement the usual method of detecting auto-antibodies in diabetes. PMID- 8400893 TI - Suppression of acute experimental allergic encephalomyelitis by the synthetic sex hormone 17-alpha-ethinylestradiol: an immunological study in the Lewis rat. AB - Induction of experimental allergic encephalomyelitis (EAE) in female Lewis rats led to the well-known clinical symptoms and histological signs. Treatment with the synthetic estrogen 17-alpha-ethinylestradiol (EE) from day -4 before induction until day 21 after induction resulted in partial suppression of these signs and symptoms. Analysis of the peripheral blood leukocyte (sub)populations in these treated animals indicated some remarkable changes. However, these changes were also observed without EE treatment. EE treatment of EAE rats resulted in a significant decrease of the relative weights of both thymus and spleen, which changes however were not reflected in the peripheral blood. Apparently the effects of EE treatment on EAE in the present experiments indicate an action locally at the site of the EAE lesion and do not seem to be mediated by gross changes in the levels of peripheral blood leukocytes. PMID- 8400894 TI - Lymphocyte proliferation induced by pertussis toxin utilizes a pathway parallel to transforming growth factor-beta-sensitive growth. AB - Pertussis toxin (PTX) has been shown to potentiate autoimmunity in experimental autoimmune disease. The exact mechanism of this effect has not been determined; however, the modification of G proteins by ADP-ribosylation has been suggested. Here it is demonstrated that this modification may contribute to autoimmunity by the abrogation of transforming growth factor-beta (TGF-beta) growth-inhibitory signals. Anti-TGF-beta demonstrated the same effect on lymphocytes as high concentrations of PTX in vitro. PMID- 8400895 TI - Properties of tree and grass pollen allergens: reinvestigation of the linkage between solubility and allergenicity. AB - In this study we reinvestigated the kinetics of allergen release from birch pollen (Betula verrucosa) and timothy grass pollen (Phleum pratense) using different protein extraction procedures, immunoblotting with specific antibodies and immune electron microscopy. Pollen allergens such as the major birch pollen allergen, Bet v I, the major timothy grass pollen allergens, Phl p I and Phl p V, group-II/III allergens from timothy grass and profilins were released rapidly and in large amounts from hydrated pollen. Within a few minutes pollen allergens could be detected in aqueous supernatants prepared from birch and grass pollen with serum IgE or specific antibodies. In parallel the allergen content in the pollen pellet fractions decreased. A nonallergenic protein such as heat shock protein 70 can be extracted in sufficient amounts only with harsh extraction procedures. Immune electron microscopy of dry and rehydrated birch pollens showed that after short hydration, the major birch pollen allergen, Bet v I, migrated into the exine and to the surface of intact pollen grains, whereas profilin, against which a lower percentage of patients is sensitized, was retained in the pollen grain. Comparing the amino acid composition and hydrophilicity of the tested allergens with a nonallergenic protein such as heat shock protein 70, no significant difference was noted. In agreement with earlier observations we conclude that the allergenic properties of proteins are rather linked to the amount and speed of solubility from airborne particles than to intrinsic properties. PMID- 8400896 TI - Purification and identification of polyclonal IgE antibodies from ragweed sensitized dog sera. AB - We have purified and characterized polyclonal dog IgE. Serum IgE was precipitated by (NH4)2SO4 and then purified by two different procedures. Ion exchange on DEAE Sephacel, followed by HPLC using Tonen hydroxylapatite and then Protein G Sepharose, produced a highly purified IgE fraction (No. 1) free of IgG, IgA and IgM as measured by ELISA, but recovery of IgE as measured by passive cutaneous anaphylaxis was low. Gel filtration on Sephacryl S-300, Con A-Sepharose and Protein G-Sepharose recovered 18% of initial IgE, 0.02% IgG, 0.4% IgM and 0.3% IgA. This IgE fraction (No. 2) was used to induce antibody production in rabbits. Western blot analysis was then performed for dog IgE fractions No. 1 and 2. Using the rabbit anti-dog IgE, a prominent IgE band with an apparent molecular mass of 226 kD was identified in fractions No. 1 and 2 subjected to nonreducing SDS-PAGE. This band also reacted with anti-human IgE, but not with anti-dog IgG or anti-dog IgA. Under reducing conditions the approximate molecular mass for the IgE & chain, estimated by Western blot using rabbit anti-dog IgE, was 73 kD, providing a molecular mass of 196 kD for dog IgE. PMID- 8400897 TI - Potentiation of IgE responses to third-party antigens mediated by Ascaris suum soluble products. AB - A reductive approach was used to examine the potentiation of IgE responses by nematode infection. Ascaris homogenized extract, Ascaris pseudocoelomic (body) fluid (ABF) and purified Ascaris allergen (ABA) were tested for their ability to act as protein carriers and as mediators of potentiated IgE responses to third party (ovalbumin; OVA) responses. All three nematode products were excellent protein carriers for the hapten dinitrophenol and showed significantly better activity in this respect than OVA. Neither ABF nor ABA enhanced the level of the IgE response to the third-party antigen but both prolonged the response markedly. ABF, but not ABA, induced high levels of total circulating IgE when given at the same time as OVA with alum. The data suggest that the enhancement and prolongation of IgE responses by nematodes may be two separate but related activities. PMID- 8400898 TI - Acquired resistance to Schistosoma japonicum in IgE-deficient SJA/9 mice immunized with irradiated cercariae. AB - Participation of IgE in protective immunity against Schistosoma japonicum was examined by comparing congenital IgE-deficient SJA/9 and IgE-producing SJL/J mice. Mice immunized with 100 irradiated cercariae 7 weeks previously were infected with 50 live cercariae. In SJL/J mice at 40 days after infection, a 3-to 4-fold increase of total IgE levels and anti-S, japonicum egg IgE antibody production were observed with no significant difference between immunized and nonimmunized mice. IgE was not detected in SJA/9 mice throughout the experiments. Protective immunity evaluated by recovery of adult worms was found in SJA/9 mice and was comparable to that of SJL/J mice. These results suggest that acquired immunity in mice with irradiated cercariae of S. japonicum was not dependent on IgE in these strains of mice. PMID- 8400899 TI - Role of capsaicin-sensitive sensory nerve reflexes in guinea pig model of nasal allergy. AB - The aim of this study was to examine the pathophysiological role of capsaicin sensitive sensory nerves in an animal model of nasal allergy. In ovalbumin (OA) sensitized guinea pigs, a significant increase in nasal total airway resistance (TAR) was noted for at least 180 min after topical antigen challenge. The TAR response to antigen challenge was significantly inhibited for 120 min by general capsaicin pretreatment (167 +/- 12.1 vs. 113 +/- 5.0%, p < 0.001, and 186 +/- 14.9 vs. 119 +/- 6.6%, p < 0.001, control vs. capsaicin pretreatment group at 20 and 90 min after challenge, respectively). However, TAR was significantly though slightly affected even after general capsaicin pretreatment. Following nasal capsaicin challenge, TAR increased for 90 min, and nasal secretion for 30 min. Both the TAR and secretory responses to nasal capsaicin challenge were significantly greater in OA-sensitized guinea pigs than in nonsensitized animals (171 +/- 12.1 vs. 137 +/- 7.4% at 30 min, p < 0.05, and 82.3 +/- 8.6 vs. 13.4 +/- 1.7 mg/10 min, p < 0.05, TAR and secretory response to 300 microM nasal capsaicin challenge, respectively). These results suggest that capsaicin-sensitive sensory nerve reflexes play an important role in the occurrence of early-phase nasal symptoms following topical antigen exposure and are accelerated in OA-sensitized guinea pigs. PMID- 8400900 TI - Dissociation of allergenic and immunogenic functions in contact sensitivity to para-phenylenediamine. AB - Contact sensitivity to para-phenylenediamine (PPD) is a frequent delayed-type hypersensitivity resulting in contact dermatitis. The aim of the present study, conducted in 16 patients allergic to PPD (as assessed by a positive patch test), was to get better insight into the mechanism of T-cell activation in PPD contact sensitivity. PPD was unable to induce significant proliferation of T cells from a first set of 9 patients. In 7 further patients, lymphocyte proliferation was assessed using PPD and 2 PPD metabolites, namely Brandrowski's base (BB) and benzoquinone (BQ). BB specifically stimulated T-cell proliferation in a dose dependent fashion in all 7 patients whereas BQ, like PPD, was ineffective. The peripheral blood mononuclear cells (PBMC) of 8 PPD nonallergic individuals did not respond to either PPD, BB or BQ. We concluded from this study that: (1) the immunogenic hapten in PPD hypersensitivity is not PPD itself, and (2) BB might be the oxidative derivative of PPD endowed with T-cell-activating properties. Further support to this statement was provided by the observation that a T cell line derived from PBMC of a PPD-allergic patient in the presence of PPD responded to BB but not to PPD. Our in vitro results suggest that PPD is a prohapten which when applied on the skin is metabolized and converted into products (such as BB) which are the immunogenic haptens able to activate specific T cells. PMID- 8400901 TI - [Polychlorinated biphenyls as a cause of sterility in women?]. AB - A total of 59 female infertile patients were investigated for the concentration of polychlorinated biphenyls (PCB) in their blood serum and follicular fluid. The average concentration of PCB in 66 related samples of follicular fluid and blood serum was 1.6 and 1.8 ng/ml, respectively. A splitting of the patients into two collectives, one with explained infertility, the other with unexplained infertility, showed no evidence of a higher PCB concentration in unexplained infertility patients. These results may exclude PCB as a primary reason for unexplained infertility. PMID- 8400902 TI - [Tuberculosis of the breast]. AB - The case of a 89-year-old women with mammary tuberculosis is reported. The diagnosis was based on histopathological and bacteriological examinations. Chest X-ray showed a lung-pleural tuberculosis, so we considered it a secondary organ manifestation. PMID- 8400903 TI - ["On the abdominal wall visible limit between the uterine body and cervix in labor". In memory of Ludwig Bandl (100th anniversary of his death)]. AB - Ludwig Bandl, who is nearly forgotten today, was one of the most active gynaecologists in the 70s and 80s of the last century. We owe him the observation and the description of the uterine contraction ring at birth. After a promising start of his career in Vienna he was professor in Prague for only 20 days. He died mentally ill in Dobling, Vienna in 1892. PMID- 8400904 TI - [Erotic aspects of the physician-patient relationship in gynecology]. PMID- 8400905 TI - [The Vorarlberg model of disease prevention]. PMID- 8400906 TI - [The role of colposcopy for cervix cancer prevention]. PMID- 8400907 TI - [Prevention and early detection of ovarian cancer]. PMID- 8400908 TI - [Prevention of breast cancer]. PMID- 8400909 TI - [Quality assurance in obstetrics and gynecology]. PMID- 8400910 TI - [Quality assurance. A review with special reference to surgical gynecology]. AB - The aim of quality assurance should be to ameliorate the medical treatment. Also it is automatically part of the medical profession. This paper defines the terms quality assurance and quality. Concepts existing in the USA and in Germany are partly just formed in Switzerland or work at the ASF (Arbeitsgemeinschaft Schweizerischer Frauenkliniken) successfully since 10 years. The quality assurance in hospitals and in the health system should be reached throughout Switzerland. PMID- 8400911 TI - [Magnesium in obstetrics and gynecology]. AB - Pregnancy induces a significant decrease of magnesium levels in serum and myometrium of 15%. A 25% increase of magnesium excretion in urine may be the most important reason. Magnesium supplementation during pregnancy seems necessary because it improves maternal health and fetal outcome. Magnesium supplementation reduces the incidence of preterm labour and vaginal haemorrhage. Premature delivery is significantly reduced from 8.2 to 2.8%. Intravenous magnesium application in pharmacological doses is still the therapeutic basis in pre eclampsia and eclampsia. In gynaecology magnesium relieves premenstrual mood changes and alleviates dysmenorrhoea. PMID- 8400912 TI - [Magnesium content of serum in pregnancy-induced hypertension]. AB - Magnesium concentrations were measured in the serum of 159 pregnant women, 19 patients with severe, 64 with mild gestational hypertension and 76 with a normal pregnancy. Our data do not support the conclusion that magnesium deficiency is the primary cause of gestational hypertension. PMID- 8400914 TI - Nursing doctoral programs in the United States. PMID- 8400913 TI - [Surgical strategy in incontinence and prolapse]. AB - By means of 5 typical examples, repeated problems in the operative strategy of incontinence and descensus are discussed. Prior to each vaginal repair a urodynamic examination should be done to clarify the risk of incontinence. In the case of vaginal repair the periurethral structures should be preserved. In the case of abdominal colposuspension nonabsorbable suture material should be used. A wide elevation should be avoided. Continent patients with a cystocele and a urodynamically verified, hidden stress incontinence need, in addition, a vaginal colposuspension. In the case of vaginal stump prolapse the organ-saving operation (vaginal sacropexy) should be preferred to colpectomy. In the case of recurring stress incontinence, patients should preoperatively undergo an intensive local therapy, thus enabling a tension-free elevation. PMID- 8400915 TI - Sigma Theta Tau International supports nursing research dissemination. PMID- 8400916 TI - Bone mineral density in professional ballet dancers. AB - We studied the effect of physical training on the bone mineral content and soft tissue composition in 42 professional ballet dancers (17 men and 25 women). Twenty-eight of them were still actively performing and 14 had retired from professional dancing. Forty-two healthy age- and sex-matched volunteers served as controls. The bone mineral density (BMD) (g/cm2) in total body, spine, hip, arms, legs, and the proximal tibia metaphysis was measured with a Lunar DPX apparatus and the BMD of the distal forearm was measured with single-photon absorptiometry (SPA). Twelve of the now retired dancers had earlier been measured with SPA in the tibia condyle during their active career in 1975. There was no significant difference in BMD values between dancers and controls with the exception of a lower BMD in the head of the male dancers and a lower BMD in the arms of female dancers. After correcting for difference in body mass index (BMI), using analysis of covariance we found significant higher BMD in the lower extremities of female dancers and in the hip of male dancers. No correlation was found between the SPA measurements from 1975 and the corresponding measurements 15 years later. The dancers were on average 9 kg lighter but of the same height as the controls. The dancers had lower body mass index and fat content but their lean body mass was the same as the controls. PMID- 8400917 TI - Concentration and hydroxyapatite binding capacity of plasma osteocalcin in chronic alcoholic men: effect of a three-week withdrawal therapy. AB - Plasma concentrations of osteocalcin (OC) were studied in 40 chronic alcoholic men (age range: 21-56 years) before and after 3 weeks of ethanol withdrawal therapy and in 50 non-alcoholic controls selected in respect to age and sex. Plasma OC level in alcoholic subjects was significantly lower than in the controls (3.0 +/- 2.6 micrograms/l and 4.7 +/- 2.8 micrograms/l, respectively). After 21 days of withdrawal therapy, plasma OC level was significantly higher than at the day of admission (5.8 +/- 3.5 micrograms/l, P < 0.001). This level was not statistically different from that of the controls. We also demonstrated that the hydroxyapatite binding capacity of plasma OC before as well as after the withdrawal period was not different from that of the controls. The acetaldehyde adduction of purified bovine OC in vitro did not change any of its immunoactivity and hydroxyapatite binding capacity. The results emphasize the fact that the decrease of plasma osteocalcin in chronic alcoholics is reversible within 3 weeks of alcohol withdrawal and that the circulating protein seems to be similar to that present in controls. PMID- 8400918 TI - Inhibitory effect of interleukin-4 on osteoclast-like cell formation in mouse bone marrow culture. AB - Recently, interleukin 4 (IL-4) was reported to inhibit bone resorption in mouse long bone culture. To test the effect of IL-4 on the formation of osteoclast-like cells, we used a mouse bone marrow culture system that formed mononuclear and multinucleated cells with osteoclast characteristics. Recombinant mouse IL-4 dose dependently inhibited the formation of tartrate-resistant acid phosphatase positive multinucleated cells [TRAP(+)MNC] induced by 1,25(OH)2D3, PTH(1-34) or Interleukin-1 alpha (IL-1 alpha). The minimum effective inhibitory concentration was 0.01 ng/ml, and 1 ng/ml IL-4 completely inhibited TRAP(+)MNC formation. IL-4 also dose-dependently inhibited the formation of tartrate-resistant acid phosphatase-positive mononuclear cells. Treatment of cultures with IL-4 for the first or last 48 h of an 8-day culture period inhibited TRAP(+)MNC formation to the same extent, whereas IFN-gamma and calcitonin suppressed TRAP(+)MNC formation mainly at the early and the late phase, respectively. IL-4, as a macrophage fusion factor, at higher concentrations (0.1-10 ng/ml), increased formation of tartrate-resistant acid phosphatase-negative multinucleated cells [TRAP(-)MNC] with giant macrophage characteristics. The half-maximal concentrations inhibiting TRAP(+)MNC formation and stimulating TRAP(-)MNC formation were 0.05 ng/ml and 2 ng/ml, respectively. These results demonstrate that IL-4 inhibits bone resorption by inhibiting the recruitment of osteoclast precursor and formation of multinucleated osteoclast-like cells, and by stimulating the formation of macrophage polykaryons. PMID- 8400919 TI - Prostaglandin E2 increased rat cortical bone mass when administered immediately following ovariectomy. AB - To investigate the effects of ovariectomy and the simultaneous administration of prostaglandin E2 (PGE2) on rat tibial shaft cortical bone histomorphometry, thirty-five 3-month-old female Sprague-Dawley rats were either ovariectomized (OVX), or sham ovariectomy (sham-OVX). The OVX rats were divided into three groups and treated with 0, 1 and 6 mg PGE2/kg/day for 90 days. The double fluorescent labeled undecalcified tibial shaft cross sections (proximal to the tibiofibular junction) of all the subjects were used for histomorphometry analysis. No differences in cross-sectional area and cortical bone area were found between sham-OVX and OVX controls, but OVX increased marrow area, intracortical porosity area and endocortical eroded perimeter. Periosteal and endocortical bone formation rates decreased with aging yet OVX prevented these changes. These OVX-induced increases in marrow area and endocortical eroded perimeter were prevented by 1 mg PGE2/kg/day treatment and added bone to periosteal and endocortical surfaces and to the marrow cavity. At the 6 mg/kg/day dose level, PGE2-treated OVX rats increased total tissue area, cortical bone area, marrow trabecular bone area, minimal cortical width and intracortical porosity area, and decreased marrow area compared to basal, sham-OVX and OVX controls. In addition, periosteal bone formation was elevated in the 6 mg PGE2/kg/day-treated OVX rats compared to OVX controls. Endocortical eroded perimeter increased from basal and sham-OVX control levels, but decreased from OVX control levels in the 6 mg PGE2/kg/day-treated OVX rats. Our study confirmed that ovariectomy does not cause osteopenia in tibial shaft cortical bone in rats, but it does stimulate endocortical bone resorption and enlarges marrow area. The new findings from the present study demonstrate that PGE2 prevents the OVX induced increases in endocortical bone resorption and marrow area and adds additional bone to periosteal and endocortical surfaces and to marrow cavity to increase total bone mass in the tibial shaft of OVX rats when given immediately following ovariectomy. PMID- 8400920 TI - S-ketoprofen inhibits tenotomy-induced bone loss and dynamics in weanling rats. AB - The objects of this study were to determine whether S-Ketoprofen, a non-steroidal anti-inflammatory drug (NSAID), can prevent immobilization (tenotomy)-induced bone loss in weanling rats. Forty-five 4-week-old Sprague-Dawley female rats were either sham-operated or subjected to knee tenotomy and treated simultaneously with 0, 0.02, 0.1, 0.5 or 2.5 mg of S-ketoprofen/kg per day for 21 days. We then studied double-fluorescent labeled proximal tibial longitudinal sections and tibial shaft cross sections using static and dynamic histomorphometry. Less cancellous bone mass in proximal tibial metaphyses was found in tenotomized controls than in basal (36%) and sham-operated (54%) controls. This was due to the inhibition of age-related bone gain and induced bone loss due to increased bone resorption and decreased bone formation. S-ketoprofen prevented both the inhibition of age-related bone gain and the stimulation of bone loss at the 2.5 mg/kg per day dose level, while it only prevented bone loss at the 0.5 mg/kg dose levels. In cancellous bone, dynamic histomorphometry showed that S-ketoprofen prevented the tenotomy induced decrease in bone formation and increase in bone resorption. In the tibial shaft, tenotomy inhibited the enlargement of total tissue area by depressing periosteal bone formation, and thus inhibited age related cortical bone gain. S-ketoprofen treatment did not prevent this change at all dose levels, but reduced marrow cavity area to increase cortical bone area at the 0.1, 0.5 and 2.5 mg/kg per dose levels compared to tenotomy controls. However, the cortical bone area in the 0.1 and 0.5 mg dose-treated tenotomy rats was still lower than in the age-related controls. S-ketoprofen also prevented the increase in endocortical eroded perimeter induced by tenotomy. In summary, tenotomy inhibited age-related bone gain and stimulated bone loss in cancellous bone sites, and only inhibited age-related bone gain in cortical bone sites. S ketoprofen treatment at the highest dose levels prevented the changes in cancellous bone, and reduced marrow area to increase cortical bone in the tibial shafts. PMID- 8400921 TI - Anabolic responses of an adult cancellous bone site to prostaglandin E2 in the rat. AB - The objects of this study were to determine: (1) the response of a non-growing cancellous bone site to daily prostaglandin E2 (PGE2) administration; and (2) the differences in the effects of daily PGE2 administration in growing (proximal tibial metaphysis, PTM) and non-growing cancellous bone sites (distal tibial metaphysis, DTM). Seven-month-old male Sprague-Dawley rats were given daily subcutaneous injections of 0, 1, 3 and 6 mg PGE2/kg per day for 60, 120 and 180 days. The static and dynamic histomorphometric analyses were performed on double fluorescent labeled undecalcified distal tibial metaphyses (DTM). No age-related changes were found in static and dynamic histomorphometry of DTM cancellous bone between 7 and 13 months of age. The DTM of 7-month-old (basal controls) rats consisted of a 24.5 +/- 7.6%-metaphyseal cancellous bone mass, and a thick trabeculae (92 +/- 12 micron). It also had a very low tissue-base bone formation rate (3.0 +/- 7.3%/year). Exogenous PGE2 administration produced the following transient changes in a dose-response manner between zero and 60 days: (1) increased trabecular bone mass and improved architecture (increased trabecular bone area, width and number, and decreased trabecular separation); (2) increased trabecular interconnections; (3) increased bone formation parameters; and (4) decreased eroded perimeter. A new steady state with more cancellous bone mass and higher bone turnover was observed from day 60 onward. The elevated bone mass induced by the first 60 days of PGE2 treatment was maintained by another 60 and 120 days with continuous daily PGE2 treatment. When these findings were compared to those previously reported for the PTM, we found that the DTM was much more responsive to PGE2 treatment than the PTM. Percent trabecular bone area and tissue-based bone formation rate increased significantly more in DTM as compared to PTM after the 60 days of 6 mg PGE2 treatment. These observations indicate that a non-growing cancellous bone site is more responsive than growing bone site to long-term daily administration of PGE2. PMID- 8400922 TI - The benefits and pitfalls of pulse oximetry. PMID- 8400923 TI - Psychological problems and the intensive care unit. PMID- 8400924 TI - European directives on medical devices. PMID- 8400925 TI - Female genital mutilation. PMID- 8400926 TI - Retinoblastoma and low level radiation. PMID- 8400927 TI - Spinal cord stimulation and pain relief. PMID- 8400928 TI - Comparisons of therapeutic effects of levodopa, levodopa and selegiline, and bromocriptine in patients with early, mild Parkinson's disease: three year interim report. Parkinson's Disease Research Group in the United Kingdom. AB - OBJECTIVE: To determine the optimum treatment for early Parkinson's disease. DESIGN: An open, long term, prospective randomised trial conducted by the Parkinson's Disease Research Group of the United Kingdom. SETTING: 93 hospitals throughout the United Kingdom. SUBJECTS: 782 patients with early Parkinson's disease who were not receiving dopaminergic treatment. INTERVENTIONS: Patients allocated to treatment with levodopa/dopa decarboxylase inhibitor alone (arm 1), levodopa/decarboxylase inhibitor/selegiline in combination (arm 2), or bromocriptine (arm 3). MAIN OUTCOME MEASURES: Disability assessment as judged by improvement on Hoehn and Yahr, modified Webster, and North Western University disability scales. Adverse event profile and mortality ratios. RESULTS: Interim results indicate that all three treatment regimens led to improvement in baseline disabilities after 12 months' treatment and that deterioration in control was apparent by three years. No significant differences were found between the results of treatment in arm 1 and arm 2, but both were significantly more effective than bromocriptine (arm 3) and had fewer early adverse reactions. The adjusted difference (95% confidence interval) in Webster rating for arm 3 v 1 was 0.93 points (0.27 to 1.50; p = 0.0058) and for arm 3 v 2 was 1.25 points (0.61 to 1.89; p = 0.0002). The incidence of dyskinesias and motor oscillations, however, was significantly lower in arm 3 (2% and 5%, respectively) than in arm 1 (27% and 33%, respectively) and arm 2 (34% and 35%, respectively). CONCLUSIONS: As there were no marked differences in functional improvement between the three groups the choice of treatment in the early stages of Parkinson's disease may not be critical. PMID- 8400929 TI - Comparison of community based service with hospital based service for people with acute, severe psychiatric illness. AB - OBJECTIVE: To compare the burden on relatives and outcome of people treated for severe acute psychiatric illness by a community service and a traditional hospital based service. DESIGN: Follow up of patients aged 16-65 who required admission to hospital or home treatment for psychiatric illness during January 1990 to February 1991. SETTING: Two Birmingham electoral wards, Sparkbrook and Small Heath; Sparkbrook has a community based service and Small Heath a traditional hospital based service. SUBJECTS: 69 patients from Sparkbrook and 55 from Small Health. MAIN OUTCOME MEASURES: Scores on present state examination, social behaviour assessment schedule, and general health questionnaire. RESULTS: 24 (35%) of Sparkbrook patients received some treatment in hospital during the initial episodes. Relatives of Sparkbrook patients were less distressed by their burden at the initial assessment than relatives of Small Health patients (mean score 0.11 v 0.29, p < 0.01). Relatives were also more satisfied with the support they received and the treatment received by patients. More patients from Sparkbrook than Small Health were in contact with a psychiatrist (81% (95% confidence interval 71% to 91%) v 62% (44% to 68%)) and community nurse (56% (44% to 68%) v 14% (13% to 24%)) one year after the initial episode. Sparkbrook patients spent significantly fewer days in hospital during the initial episode (8 days v 59 days) and the first year (20.6 v 67.9 days). CONCLUSION: The community based service is as effective as the hospital based service and is preferred by relatives. It is more effective in keeping people in long term contact with psychiatrists. PMID- 8400930 TI - Effects of spinal cord stimulation in angina pectoris induced by pacing and possible mechanisms of action. AB - OBJECTIVE: To investigate the effects of spinal cord stimulation on myocardial ischaemia, coronary blood flow, and myocardial oxygen consumption in angina pectoris induced by atrial pacing. DESIGN: The heart was paced to angina during a control phase and treatment with spinal cord stimulation. Blood samples were drawn from a peripheral artery and the coronary sinus. SETTING: Multidisciplinary pain centre, department of medicine, Ostra Hospital, and Wallenberg Research Laboratory, Sahlgrenska Hospital, Gothenburg, Sweden. SUBJECTS: Twenty patients with intractable angina pectoris, all with a spinal cord stimulator implanted before the study. RESULTS: Spinal cord stimulation increased patients' tolerance to pacing (p < 0.001). At the pacing rate comparable to that producing angina during the control recording, myocardial lactate production during control session turned into extraction (p = 0.003) and, on the electrocardiogram, ST segment depression decreased, time to ST depression increased, and time to recovery from ST depression decreased (p = 0.01; p < 0.05, and p < 0.05, respectively). Spinal cord stimulation also reduced coronary sinus blood flow (p = 0.01) and myocardial oxygen consumption (p = 0.02). At the maximum pacing rate during treatment, all patients experienced anginal pain. Myocardial lactate extraction reverted to production (p < 0.01) and the magnitude and duration of ST segment depression increased to the same values as during control pacing, indicating that myocardial ischaemia during treatment with spinal cord stimulation gives rise to anginal pain. CONCLUSIONS: Spinal cord stimulation has an anti-anginal and anti-ischaemic effect in severe coronary artery disease. These effects seem to be secondary to a decrease in myocardial oxygen consumption. Furthermore, myocardial ischemia during treatment gives rise to anginal pain. Thus, spinal cord stimulation does not deprive the patient of a warning signal. PMID- 8400931 TI - Adverse drug reactions: who is to know? PMID- 8400932 TI - Homozygous deletion of gene for glutathione S-transferase M1 in bladder cancer. PMID- 8400933 TI - Predicting driving performance after stroke. PMID- 8400934 TI - Remission of seizures in untreated epilepsy. PMID- 8400935 TI - Agranulocytosis associated with cephalosporin. PMID- 8400936 TI - Cyclosporin induced colitis. PMID- 8400937 TI - Erythroderma after clodronate treatment. PMID- 8400938 TI - Transcutaneous overdose of terbutaline. PMID- 8400939 TI - Schizophrenia sans frontieres: concepts of schizophrenia among French and British psychiatrists. AB - OBJECTIVE: To compare the first admission rates for schizophrenia in England and France, and to compare the concept of schizophrenia held by practising British and French psychiatrists. DESIGN: Comparative study of incidence rates in England and France; and postal questionnaire survey of a sample of about 1 in 30 psychiatrists in the United Kingdom and in l'Aquitaine, France. SUBJECTS: All first admissions for schizophrenia to psychiatric hospitals in England and France 1973-82; 92 psychiatrists in the United Kingdom and 69 in France. MAIN OUTCOME MEASURES: Age adjusted first admission rates for schizophrenia between 1973-82; and opinions on the aetiology, diagnosis, and management of schizophrenia. RESULTS: First admission rates were much higher in France than in England before the age of 45, but lower after that age. Rates were falling in England over the 10 year period, while they were rising in France. In the questionnaire study English and French psychiatrists showed prominent differences of opinion for 31 out of 38 statements. The French sample did not diagnose schizophrenia after the age of 45 and endorsed psychoanalytical concepts. CONCLUSIONS: British and French psychiatrists use different diagnostic criteria and contrasting methods of treatment for schizophrenia. Differences in diagnostic criteria probably contribute towards the disparity in administrative incidence rates and time trends for schizophrenia in the two countries. Doctors in the European Community can now work in any country. Further work is needed to ensure psychiatrists are talking a common language. PMID- 8400940 TI - Epilepsy and pregnancy. PMID- 8400941 TI - A case of chylous ascites. PMID- 8400942 TI - Sexual health. Consider people, not gender. PMID- 8400943 TI - Sexual health. Male stereotyping is unhelpful. PMID- 8400944 TI - Serum screening for Down's syndrome. Informed consent is vital... PMID- 8400945 TI - Serum screening for Down's syndrome. ...but time consuming and expensive. PMID- 8400946 TI - Serum screening for Down's syndrome. Private screening is problematic. PMID- 8400947 TI - Serum screening for Down's syndrome. Widening the programme would be costly. PMID- 8400948 TI - Health of children born prematurely. Antenatal factors may be important. PMID- 8400949 TI - Health of children born prematurely. New treatment reduces risk of blindness. PMID- 8400950 TI - Serum lipid testing. PMID- 8400951 TI - How can doctors diagnose colorectal cancer earlier? PMID- 8400952 TI - Subcortical dementia. PMID- 8400953 TI - Chloroquine poisoning. PMID- 8400954 TI - Citizens' advice in general practice. PMID- 8400955 TI - Videotaping general practice consultations. PMID- 8400956 TI - Treatments for tinnitus. PMID- 8400957 TI - Acute circulatory support. Noradrenaline may preserve renal function. PMID- 8400958 TI - Sex ratio errors in census data. PMID- 8400959 TI - Vaccine given bad press. PMID- 8400960 TI - Sickle cell disease and contraception. PMID- 8400961 TI - Microalbuminuria and diabetes. PMID- 8400962 TI - Like father like son. PMID- 8400963 TI - Clubbing in Crohn's disease. PMID- 8400964 TI - Schizophrenic patients requiring intensive care. PMID- 8400965 TI - Primary care for children. Illustrations perpetuate bad practice. PMID- 8400966 TI - Hyperbaric oxygen. PMID- 8400967 TI - Primary care and public health. PMID- 8400968 TI - Creutzfeldt-Jakob disease after pituitary gonadotrophins. PMID- 8400969 TI - The role of doctors in promoting smoking cessation. PMID- 8400970 TI - Steroid osteoporosis. PMID- 8400971 TI - Meta-analysis of randomised controlled trials of selective decontamination of the digestive tract. Selective Decontamination of the Digestive Tract Trialists' Collaborative Group. AB - OBJECTIVE: To determine the clinical benefits of selective decontamination of the digestive tract in patients treated in intensive care units. DESIGN: Meta analysis of 22 randomised trials that compared different combinations of oral non absorbable antibiotics, with or without a systemic component, with no treatment in controls. SUBJECTS: 4142 patients seen in general and specialised intensive care units around the world. 2047 received some form of antibiotic treatment, the remainder no prophylaxis. DATA ANALYSIS: Each trial was reviewed through direct contact with study investigators. Data collected were: the randomisation procedure, number of patients, number excluded from the analysis, and numbers of respiratory tract infections and deaths. Data were combined according to an intention to treat analysis with the Mantel-Haenszel-Peto method. MAIN OUTCOME MEASURES: Respiratory tract infections and total mortality. RESULTS: Selective decontamination of the digestive tract significantly reduced respiratory tract infections (odds ratio 0.37; 95% confidence interval 0.31 to 0.43). The value of the common odds ratio for total mortality (0.90; 0.79 to 1.04) suggested at best a moderate treatment effect, reaching statistical significance only when the subgroup of trials of topical and systemic treatment combined was considered separately (odds ratio 0.80; 0.67 to 0.97). No firm conclusions could be drawn owing to large variations in patient mix and severity within and between trials. CONCLUSIONS: The findings strongly indicate that selective decontamination significantly reduces infection related morbidity in patients receiving intensive care. They also highlight why definite conclusions about the effect of prophylaxis on mortality cannot be drawn despite the large number of trials available. Based on the most favourable results obtained by pooling data from trials in which combined topical and systemic treatment was used it may be estimated that 6 (range 5-9) and 23 (13-139) patients would need to be treated to prevent one respiratory tract infection and one death respectively. PMID- 8400972 TI - Termination of pregnancy with reduced doses of mifepristone. World Health Organisation Task Force on Post-ovulatory Methods of Fertility Regulation. AB - OBJECTIVES: To compare the abortifacient efficacy and side effects of three doses of the antiprogestin mifepristone plus prostaglandin for termination of early pregnancy. DESIGN: Randomised, double blind multicentre trial. SETTING: 11 departments of obstetrics and gynaecology and of family planning, mostly in university hospitals, in seven countries. SUBJECTS: 1182 women with an early pregnancy (menstrual delay of 7-28 days) requesting abortion. INTERVENTIONS: Single doses of 200 mg, 400 mg, or 600 mg mifepristone followed, 48 hours later, by vaginal pessary of 1 mg of the prostaglandin E1 analogue gemeprost. MAIN OUTCOME MEASURES: Outcome of treatment; duration and subjective amount of menstrual bleeding; side effects and complications; and concentrations of haemoglobin. RESULTS: Outcome was similar with the three doses of mifepristone. Of the 1151 women with known outcome, 95.5% had a complete abortion (364 (93.8%) of those given 200 mg mifepristone, 368 (94.1%) of those given 400 mg, and 367 (94.3%) of those given 600 mg), 3.7% had an incomplete abortion (14 (3.6%), 15 (3.8%), and 14 (3.6%)), 0.3% had a missed abortion (three (0.8%), one (0.3%), and none), and 0.4% had a continuing live pregnancy (two (0.5%), two (0.5%), and one (0.3%)). Of the 43 women who had incomplete abortion, 23 underwent emergency uterine curettage (usually for haemostatic purposes) and three of these women were given a blood transfusion. The numbers of reported complaints, bleeding patterns, and changes in blood pressure and haemoglobin concentrations were similar with the three treatments. CONCLUSIONS: For termination of early pregnancy a single dose of 200 mg mifepristone is as effective as the currently recommended dose of 600 mg when used in combination with a vaginal pessary of 1 mg gemeprost. PMID- 8400973 TI - Obesity as a determinant for response to antihypertensive treatment. AB - OBJECTIVE: To test the hypothesis that beta blockers lower blood pressure more effectively than calcium entry blockers in obese hypertensive patients and that calcium entry blockers are more effective in lean patients. DESIGN: Double blind, randomised controlled trial of treatment over six weeks. SETTING: Tertiary referral centre. SUBJECTS: 42 white men with uncomplicated mild to moderate essential hypertension (World Health Organisation stage I or II); 36 completed the study. INTERVENTION: Patients were randomised to metoprolol 50-100 mg twice daily or isradipine 2.5-5.0 mg twice daily for six weeks after a two week run in phase. MAIN OUTCOME MEASURE: Blood pressure after six weeks of treatment. RESULTS: When stratified according to treatment and presence of obesity (body mass index < or = 27 kg/m2), the mean (SD) fall in blood pressure in the beta blocker group was 24 (13)/18 (10) mm Hg in obese patients and 18 (19)/12 (13) mm Hg in lean patients. In the calcium entry blocker group, the fall in blood pressure was 21 (15)/17 (6) mm Hg in lean patients and 18 (11)/8 (10) mm Hg in obese patients. After taking age and blood pressure before treatment into account there was a significant interaction between obesity and drug therapy (p = 0.019) with a better diastolic blood pressure response to calcium entry blockers in lean patients and to beta blockers in obese hypertensive patients. CONCLUSION: Obesity affects the efficacy of metoprolol and isradipine in reducing blood pressure. PMID- 8400974 TI - Kidney granuloma in Crohn's disease. PMID- 8400975 TI - Post-tropical screening: how useful is it? PMID- 8400976 TI - MRCGP pass rate by medical school and region of postgraduate training. Royal College of General Practitioners. PMID- 8400977 TI - Distribution of mental health professionals working on site in English and Welsh general practices. AB - OBJECTIVE: To describe the nature and distribution of mental health professionals working on site in general practices. DESIGN: Postal questionnaire and telephone interview survey. SETTING: English and Welsh general practices. SUBJECTS: 1880 general practitioners, of whom 1542 (82%) responded. MAIN OUTCOME MEASURES: Prevalence, types, and distribution of mental health professionals working on site among general practices. Factors predicting the presence of mental health professionals on site. RESULTS: The number of practices reporting the presence on site of each type of professional were 528 for community psychiatric nurses; 266 for practice counsellors; 177 for clinical psychologists; 132 for psychiatrists; 96 for psychiatric social workers; and 45 for psychotherapists. Mental health professionals tended to cluster together in practices more often than expected by chance alone. Practice characteristics which independently predicted the presence of a mental health professional on site were having four or more partners; being a training practice; and running stress, bereavement, or other mental health clinics. The proportions of practices with mental health professionals on site varied significantly among health regions. There was no association between the presence of mental health professionals on site and the location of practices, the social class mix of patients, or the estimated percentage of elderly patients or patients of non-European origin. CONCLUSIONS: Mental health professionals tend to cluster together, with a preponderance in larger training practices. Specialist mental health care provision within general practices is unevenly distributed. Further research is needed to determine whether this uneven distribution reflects differences in need or inequalities in the provision of mental health services. PMID- 8400978 TI - Guidelines for resuscitation and transfer of patients with serious head injury. PMID- 8400979 TI - Write a departmental handbook for junior staff. PMID- 8400980 TI - Lesions of schistosomiasis mimicking warts on the vulva. PMID- 8400981 TI - Resource restraints: what do we tell our patients? PMID- 8400982 TI - Routine ultrasound scanning in pregnancy. The benefits are clinical... PMID- 8400983 TI - Routine ultrasound scanning in pregnancy. Apgar scores are poor predictors of outcome. PMID- 8400984 TI - Craniosynostosis. PMID- 8400985 TI - Heparin induced thrombosis. May complicate renal replacement treatment. PMID- 8400986 TI - Annual influx of temporary residents. To the Glastonbury Festival... PMID- 8400987 TI - Heparin induced thrombosis. Datasheet warns of risk. PMID- 8400988 TI - Hepatitis A virus infection. Risk to sewage workers unproved. PMID- 8400989 TI - Hepatitis A virus infection. No conclusive link to factor VIII. PMID- 8400990 TI - Hepatitis A virus infection. Should susceptible homosexual men be offered immunization. PMID- 8400991 TI - The Hawksley random zero sphygmomanometer. Comparison with mercury instrument is illogical. PMID- 8400992 TI - Preventing neurocysticercosis. PMID- 8400993 TI - Ultrasonography and handedness. Don't confuse direction with degree. PMID- 8400994 TI - Fatigue in general practice. A difficult subject for study. PMID- 8400995 TI - Fatigue in general practice. Sex difference exaggerated. PMID- 8400996 TI - Chloroquine overdose in Papua New Guinea. PMID- 8400997 TI - Paediatric intensive care. PMID- 8400998 TI - Confidentiality for doctors with AIDS. Guidelines must be reformed. PMID- 8400999 TI - Confidentiality for doctors with AIDS. Official sources maintained confidentiality. PMID- 8401000 TI - Availability of specialist services. Deaf people need psychiatric services. PMID- 8401001 TI - Deciding who needs dual chamber pacing. PMID- 8401002 TI - NSAIDs in the postoperative period. PMID- 8401003 TI - Health care in South Africa. Don't dismiss the private option. PMID- 8401004 TI - Health care in South Africa. Rural areas suffer most. PMID- 8401005 TI - Psychological survival after concentration camps. PMID- 8401006 TI - Reference management software. Libraries can help you... PMID- 8401007 TI - Risk factors for death in men with glucose intolerance. PMID- 8401008 TI - Iron and coronary heart disease. PMID- 8401009 TI - From specialist care to self directed treatment. PMID- 8401010 TI - Apheresis in the 1990s. PMID- 8401011 TI - Leukaemia: a genetic disorder of haemopoietic cells. PMID- 8401012 TI - Argentinian doctors accused of spreading AIDS. PMID- 8401013 TI - Influence of oral magnesium supplementation on cardiac events among survivors of an acute myocardial infarction. AB - OBJECTIVE: To investigate the effect of long term oral magnesium treatment on incidence of cardiac events among survivors of an acute myocardial infarction. DESIGN: Double blind, placebo controlled parallel study in which patients were randomised to treatment or placebo. SETTING: Two coronary care units and corresponding outpatient clinics. SUBJECTS: 468 survivors of an acute myocardial infarction (289 men and 178 women) aged 31-92. INTERVENTIONS: One tablet of 15 mmol magnesium hydroxide or placebo daily for one year. MAIN OUTCOME MEASURES: Incidences of reinfarction, sudden death, and coronary artery bypass grafting in one year. RESULTS: There was no significant difference between treatment and placebo groups in the incidence of each of the three cardiac events, but when the events were combined and drop outs were excluded from calculations there was a significantly higher incidence of events in the treatment group (56/167 v 33/153; relative risk 1.55 (95% confidence interval 1.07 to 2.25); p = 0.02). When the timing of events was incorporated by means of a Kaplan-Meier plot the treatment group showed a significantly higher incidence of events whether drop outs were included or excluded (p < 0.025). CONCLUSION: Long term oral treatment with 15 mmol magnesium daily doses not reduce the incidence of cardiac events in survivors of an acute myocardial infarction and, indeed, seems to increase the risk of developing a cardiac event. Consequently, this treatment cannot be recommended as secondary prophylaxis for such patients. PMID- 8401014 TI - Birth weight from pregnancies dated by ultrasonography in a multicultural British population. AB - OBJECTIVE: To produce standard curves of birth weight according to gestational age validated by ultrasonography in the British population, with particular reference to the effects of ethnic origin. DESIGN: Retrospective analysis of computerised obstetric database. SETTING: Three large maternity units associated with Nottingham University with over 16,000 deliveries a year. PATIENTS: 41,718 women with ultrasound dated singleton pregnancies and delivery between 168 and 300 days' gestation. MAIN OUTCOME MEASURES: Length of gestation, ethnic origin, parity, maternal height and weight at booking, smoking during pregnancy; the effect of these variables on birth weight. RESULTS: Birth weights from ultrasound dated pregnancies have a higher population mean and show less flattening of the birthweight curve at term than those of pregnancies dated from menstrual history. Significant differences were observed in mean birth weights of babies of mothers of European origin (3357 g), of Afro-Caribbean origin (3173 g), and from the Indian subcontinent (3096 g). There were also significant interethnic differences in length of gestation, parity, maternal height, booking weight, and smoking habit which affected birth weight. The ethnic differences in birth weight were even greater when the effect of smoking was excluded. CONCLUSIONS: Birthweight standards require precise dating of pregnancy and should describe the population from which they were derived. In a heterogeneous maternity population the accurate assessment of an individual baby's weight needs to take the factors which affect birthweight standards into consideration. PMID- 8401015 TI - Gastric cancer: a curable disease in Britain. AB - OBJECTIVE: To determine whether more vigorous efforts aimed at earlier diagnosis allied to radical surgical resection lead to improved survival of patients with gastric cancer. DESIGN: Prospective audit of all cases of gastric cancer treated during 1970-89. SETTING: Department of surgery, general hospital. SUBJECTS: 493 consecutive patients with gastric adenocarcinoma. MAIN OUTCOME MEASURES: Operative mortality, postoperative morbidity, and five year survival after radical potentially curative resection. RESULTS: 207 (42%) patients underwent potentially curative resection. The proportion of all patients in whom this was possible increased significantly (p < 0.01) from 31% in the first five year period to 53% in the last five year period. The proportion of patients who had early gastric cancer rose from 1% to 15% (p < 0.01) and stage I disease rose from 4% to 26% (p < 0.001). After potentially curative resection, mortality 30 days after operation was 6%. Operative mortality decreased from 9% in the 1970s to 5% in the 1980s. Likewise, the incidence of serious postoperative complications decreased from 33% in the 1970s to 17% in the 1980s (p < 0.01). Five year survival was 60% in patients who underwent curative resection, 98% in patients with early gastric cancer, and 93%, 69%, and 28% in stage I, II, and III disease respectively. By the late 1980s five year survival after operation was about 70%. CONCLUSIONS: These findings suggest that an increasing proportion of patients with gastric cancer could be diagnosed at a relatively early pathological stage when about two thirds are curable by means of radical surgery. PMID- 8401016 TI - Psychosocial predisposing factors for infantile colic. AB - OBJECTIVE: To study associations between characteristics of families during the first pregnancy and after childbirth and the development of infantile colic. DESIGN: Randomised, stratified cluster sampling. Follow up from the first visit to a maternity health care clinic during pregnancy to three months after birth with confidential semistructured questionnaires. SETTING: Maternity health care clinics in primary health care centres in Finland. SUBJECTS: 1443 nulliparous women and 1407 partners. Altogether 1333 women and 1279 men returned the questionnaires. When the infants were 3 months old 1208 women and 1115 men returned questionnaires. MAIN OUTCOME MEASURES: Marital relationship; personal and social behaviour of parents during the pregnancy and their coping with the pregnancy; mothers' physical health and events, symptoms, and experiences in relation to pregnancy; self confidence and experiences of mothers and fathers in relation to childbirth; and parents' sociodemographic and educational variables. Measure of colic when the infant was 3 months old. RESULTS: Experience of stress and physical symptoms during the pregnancy, dissatisfaction with the sexual relationship, and negative experiences during childbirth were associated with the development of colic in the baby. None of the sociodemographic factors was associated with colic. CONCLUSIONS: Early preventive health work during pregnancy should attempt to improve parents' tolerance of symptoms of stress and ability to cope and increase their confidence in parenting abilities. PMID- 8401017 TI - Changes in ethnic minority membership of health authorities 1989-92. PMID- 8401018 TI - A survey of intravenous drug administration by preregistration house officers. PMID- 8401019 TI - Compliance with standardised assessment scales for elderly people among consultant geriatricians in Wessex. PMID- 8401020 TI - The views of singlehanded general practitioners: a qualitative study. AB - OBJECTIVES: To examine the concerns of singlehanded general practitioners working in an inner London area and to compare the views of general practitioners in partnerships. DESIGN: Qualitative analysis of semistructured interviews with a random sample of singlehanded general practitioners and a sample of general practitioners from partnerships matched for age and sex. SETTING: The area covered by Lambeth, Southwark, and Lewisham Family Health Services Authority. RESULTS: The singlehanded general practitioners were more likely to be older, male, and first qualified abroad than general practitioners in partnerships. Their major concerns were inadequate premises, maintaining their singlehanded status, and coping with recent changes to their contract. Most were very satisfied with their solo status and did not see the provision of 24 hour care as stressful. CONCLUSION: Singlehanded general practitioners saw themselves as providing a unique service for patients, and their status as an alternative for general practitioners who were unhappy in partnerships. Such practices are unlikely to wither away as a pattern of provision. Any comprehensive development of primary care must take their needs into account. PMID- 8401021 TI - Ethical use of covert videoing techniques in detecting Munchausen syndrome by proxy. AB - Munchausen syndrome by proxy is an especially malignant form of child abuse in which the carer (usually the mother) fabricates or exacerbates illness in the child to obtain medical attention. It can result in serious illness and even death of the child and it is difficult to detect. Some investigators have used video to monitor the carer's interaction with the child without obtaining consent -covert videoing. The technique presents several ethical problems, including exposure of the child to further abuse and a breach of trust between carer, child, and the professionals. Although covert videoing can be justified in restricted circumstances, new abuse procedures under the Children Act now seem to make its use unethical in most cases. Sufficient evidence should mostly be obtained from separation of the child and carer or videoing with consent to enable action to be taken to protect the child under an assessment order. If the new statutory instruments prove ineffective in Munchausen syndrome by proxy covert videoing may need to be re-evaluated. PMID- 8401022 TI - Travel medicine and general practice: a suitable case for audit? AB - Travel medicine becomes more important with the continual expansion of international travel and the increased popularity of exotic holiday destinations. In the United Kingdom general practitioners provide the bulk of travel health advice and immunisation and there is growing interest in providing these services. While their armamentarium has been expanded with attractive but expensive new vaccines, the need for health service advice has never been more vital, with the risks of HIV infection and drug resistant malaria. Advantages of a general practice based travel medicine service include maintaining continuity of care for the patient, but a disadvantage might be that the general practitioner sees too few patients to acquire enough skill in the subject. Furthermore, there may be a conflict of interest between time devoted to the "vaccination ritual" and giving health advice. Overall there seems to be a case for both audit and support by the health authorities. PMID- 8401023 TI - Soft tissue injury of the cervical spine. Consider the nature of the accident. PMID- 8401024 TI - Soft tissue injury of the cervical spine. Discuss investigations with a radiologist. PMID- 8401025 TI - Transmission of HIV in prison. Prisoners need condoms and clean needles. PMID- 8401026 TI - Transmission of HIV prison. Prevention depends on enlightened approach. PMID- 8401027 TI - Pulmonary microembolisation and temazepam. PMID- 8401028 TI - Complementary medicine. Acupuncture has weak scientific foundations. PMID- 8401029 TI - Complementary medicine. Orthodox medicine isn't infallible. PMID- 8401030 TI - Complementary medicine. Elderly people use complementary treatments. PMID- 8401031 TI - Complementary medicine. BMA pushes for more research. PMID- 8401032 TI - Management of patients in persistent vegetative state. Tony Bland had intensive treatment. PMID- 8401033 TI - Triazolam. Narrow safety margin is unacceptable. PMID- 8401034 TI - Triazolam. Small studies miss the difference. PMID- 8401035 TI - Triazolam. The evidence against is extensive and consistent. PMID- 8401036 TI - Cardiopulmonary resuscitation. Guidelines should be reconsidered. PMID- 8401037 TI - Cardiopulmonary resuscitation. Bicarbonate is of no value. PMID- 8401038 TI - Infertility information pack for practitioners. PMID- 8401039 TI - The Calman report. Should lead to consultant based service. PMID- 8401040 TI - Thousands starving in Liberia. PMID- 8401041 TI - New Zealand's health care reforms. PMID- 8401042 TI - Faecal incontinence. PMID- 8401043 TI - Organs from animals. PMID- 8401044 TI - Intraoperative use of thrombolytic agents. PMID- 8401045 TI - The fish odour syndrome. PMID- 8401046 TI - Drugs: from prescription only to pharmacy only. PMID- 8401047 TI - Widow given aid to sue over use of zidovudine. PMID- 8401048 TI - Comparison of terbinafine and clotrimazole in treating tinea pedis. AB - OBJECTIVE: To compare the efficacy and safety of terbinafine 1% cream and clotrimazole 1% cream in the treatment of tinea pedis. DESIGN: Multicentre, double blind parallel group study. SETTING: 32 general practices and one hospital. PATIENTS: 256 patients with mycologically confirmed tinea pedis. Of the 211 patients evaluable, 107 were randomised to terbinafine (75 male, 32 female; mean (range) age 40 (12-81) years) and 104 to clotrimazole (79 male, 25 female; mean (range) age 36 (12-71) years). INTERVENTIONS: Terbinafine 1% cream applied twice daily for one week and inert cream applied twice daily for the next three weeks. Clotrimazole 1% cream applied twice daily for four weeks. MAIN OUTCOME MEASURES: Mycological cure (negative results on microscopy and culture) and effective treatment (mycological cure plus no or minimal signs and symptoms) measured at weeks 1, 2, 3, 4, and 6. RESULTS: At week four rates of mycological cure were 93.5% for terbinafine and 73.1% for clotrimazole (p = 0.0001); and at week six 97.2% for terbinafine and 83.7% for clotrimazole (p = 0.001). Rates of effective treatment at week 4 were 89.7% for terbinafine and 58.7% for clotrimazole (p = 0.0001); and 89.7% for terbinafine and 73.1% for clotrimazole (p = 0.002) at week 6. CONCLUSION: These results indicate that a one week course of terbinafine 1% cream is more effective in the treatment of tinea pedis than a four week course of clotrimazole 1% cream, both in terms of mycological cure and effective treatment. PMID- 8401049 TI - Psychiatric consequences of road traffic accidents. AB - OBJECTIVE: To determine the psychiatric consequences of being a road traffic accident victim. DESIGN: Follow up study of road accident victims for up to one year. SETTING: Emergency department of the John Radcliffe Hospital, Oxford. SUBJECTS: 188 consecutive road accident victims aged 18-70 with multiple injuries (motorcycle or car) or whiplash neck injury, who had not been unconscious for more than 15 minutes, and who lived in the catchment area. MAIN OUTCOME MEASURES: Present state examination "caseness"; post-traumatic stress disorder and travel anxiety; effects on driving and on being a passenger. RESULTS: Acute, moderately severe emotional distress was common. Almost one fifth of subjects, however, suffered from an acute stress syndrome characterised by mood disturbance and horrific memories of the accident. Anxiety and depression usually improved over the 12 months, though one tenth of patients had mood disorders at one year. In addition, specific post-traumatic symptoms were common. Post-traumatic stress disorder occurred during follow up in one tenth of patients, and phobic travel anxiety as a driver or passenger was more common and frequently disabling. Emotional disorder was associated with having pre-accident psychological or social problems and, in patients with multiple injuries, continuing medical complications. Post-traumatic syndromes were not associated with a neurotic predisposition but were strongly associated with horrific memories of the accident. They did not occur in subjects who had been briefly unconscious and were amnesic for the accident. Mental state at three months was highly predictive of mental state at one year. CONCLUSIONS: Psychiatric symptoms and disorder are frequent after major and less severe road accident injury. Post-traumatic symptoms are common and disabling. Early information and advice might reduce psychological distress and travel anxiety and contribute to road safety and assessing "nervous shock." PMID- 8401050 TI - Factors influencing recovery from headache after common whiplash. AB - OBJECTIVE: To assess the relation between pretraumatic and trauma related headache in patients suffering from whiplash. DESIGN: Follow up study of patients examined a mean (SD) of 7.4 (4.2) days after trauma and again at three and six months. SETTING: Patients referred from primary care. SUBJECTS: 117 patients (mean age 30.8 (9.5) years. MAIN OUTCOME MEASURES: Prevalence of trauma related headache and the predictive relation by multiple logistic regression between different somatic and psychological variables and trauma related headache at each examination. RESULTS: Prevalence of trauma related headache decreased from 57% to 27%. History of pretraumatic headache proved a significant risk factor for presenting with trauma related headache. A significant relation between trauma related headache and the following variables was found: at seven days the initial wellbeing score, early onset of neck pain, depression scale from the personality inventory, and the initial intensity of neck pain; at three months, intensity of neck pain, and history of pretraumatic headache; and at six months neck pain, pain intensity, and history of pretraumatic headache. CONCLUSIONS: History of pretraumatic headache significantly increases the likelihood of presenting with trauma related headache but only in combination with findings indicative of clinically important injury to the cervical spine. PMID- 8401051 TI - The fish odour syndrome: biochemical, familial, and clinical aspects. AB - OBJECTIVES: To study the biochemical, familial, and clinical features of the fish odour syndrome among subjects with suspected body malodour. DESIGN: Subjects who responded to a newspaper article were screened for the fish odour syndrome by interview and biochemical tests. Families of subjects with the syndrome were tested if possible. SETTING: St Mary's Hospital, London, and some interviews at subjects' homes. SUBJECTS: 187 subjects (28 males) with suspected body malodour, of whom 156 (19 males) underwent biochemical tests. Five families of six of the subjects with the fish odour syndrome agreed to further tests. MAIN OUTCOME MEASURES: Amounts of trimethylamine and trimethylamine N-oxide in urine collected over 24 hours under normal dietary conditions and for eight hours after oral challenge with 600 mg trimethylamine. RESULTS: The fish odour syndrome was diagnosed in 11 subjects: the percentage of total trimethylamine excreted in their urine samples that was oxidised to trimethylamine N-oxide was < 55% under normal dietary conditions and < 25% after oral challenge with trimethylamine (in normal subjects > 80% of trimethylamine was N-oxidised). Parents of six of the subjects with the syndrome were tested: all showed impaired N-oxidation of excreted trimethylamine (< 80%) after oral challenge, indicating that they were heterozygous carriers of the allele for the syndrome. The syndrome was associated with various psychosocial reactions including clinical depression. CONCLUSIONS: The fish odour syndrome can be inherited in an autosomal recessive fashion. It should be considered as a possible causative factor in patients complaining of body malodour. PMID- 8401052 TI - Illegibility of drug ampoule labels. PMID- 8401053 TI - Value of pulsus paradoxus in assessing acute severe asthma. British Thoracic Society Standards of Care Committee. PMID- 8401054 TI - Parents' perceptions of taking babies' rectal temperature. AB - OBJECTIVE: To explore the attitudes of parents to measuring their babies' rectal temperature. DESIGN: Qualitative study using unstructured interviews of parents given "Baby Check," a scoring system designed to assess severity of illness in babies that includes measurement of rectal temperature. SETTING: One inner city general practice in Newcastle upon Tyne. SUBJECTS: 42 parents of 34 babies under 6 months old. RESULTS: Parents were reluctant to measure rectal temperature in their babies; 37 parents spontaneously raised concerns. Fifteen did not undertake measurement, 16 did so once only, and 11 did so more than once. Parents' concerns included a fear of hurting their child, anxieties about sexual intimacy and abuse, difficulty in comforting their child, and concern for their child's feelings. Most (33) substituted axillary measurement. CONCLUSIONS: Parents' preference for the axillary method of measuring temperature and resistance to using a rectal method in their children was based on several concerns. If parents are to be encouraged to use the rectal method of measuring temperature in sick babies any benefits must be set against the generation of considerable parental anxiety and the resources that would be necessary to negotiate with parents and change their views. PMID- 8401055 TI - Diagnosis and treatment of systemic lupus erythematosus. PMID- 8401056 TI - "Whistleblowing": a health issue. AB - OBJECTIVE: To examine the response of organizations to "whistleblowing" and the effects on individual whistleblowers. DESIGN: Questionnaire survey of whistleblowers who contacted Whistleblowers Australia after its publicity campaign. SETTING: Australia. SUBJECTS: 25 men and 10 women from various occupations who had exposed corruption or danger to the public, or both, from a few months to over 20 years before. RESULTS: All subjects in this non-random sample had suffered adverse consequences. For 29 victimization had started immediately after their first, internal, complaint. Only 17 approached the media. Victimization at work was extensive: dismissal (eight subjects), demotion (10), and resignation or early retirement because of ill health related to victimization (10) were common. Only 10 had a full time job. Long term relationships broke up in seven cases, and 60 of the 77 children of 30 subjects were adversely affected. Twenty nine subjects had a mean of 5.3 stress related symptoms initially, with a mean of 3.6 still present. Fifteen were prescribed long term treatment with drugs which they had not been prescribed before. Seventeen had considered suicide. Income had been reduced by three quarters or more for 14 subjects. Total financial loss was estimated in hundreds of thousands of Australian dollars in 17. Whistleblowers received little or no help from statutory authorities and only a modest amount from workmates. In most cases the corruption and malpractice continued unchanged. CONCLUSION: Although whistleblowing is important in protecting society, the typical organisational response causes severe and longlasting health, financial, and personal problems for whistleblowers and their families. PMID- 8401057 TI - Economic evaluation and health care. What does it mean? AB - Ever since the concept of value for money in health care was introduced into the NHS, economic terms and jargon have become part of our everyday lives--but do we understand what the different types of economic evaluation all mean, particularly those that sound similar to the uninitiated? This article introduces readers to the purpose of economic evaluation, and briefly explains the differences between cost-minimisation analysis (used when the outcomes of the procedures being compared are the same); cost-effectiveness analysis (used when the outcomes may vary, but can be expressed in common natural units, such as mm Hg for treatments of hypertension); cost-utility analysis (used when outcomes do vary--for example, quality of life scales); and cost-benefit analysis (used when a monetary value is being placed on services received). Further articles will deal with each one in more detail. PMID- 8401058 TI - Taking babies' temperatures: science versus social taboos in battle over Baby Check. AB - Baby Check, a scoring system for assessing the severity of illness in babies under 6 months old, has not met with the success its developers expected. The inclusion of rectal temperature in the assessment was strongly opposed by the Royal College of Midwives, which refused to alter its view despite evidence of the safety and accuracy of rectal thermometry. British parents appear not to like rectal thermometers either. Most other medical bodies have supported Baby Check and the reason for midwives' opposition may have more to do with professional pique at not being consulted than clinical wisdom. PMID- 8401059 TI - Implementing clinical guidelines. Has a lot to offer patient care. PMID- 8401060 TI - Implementing clinical guidelines. It works in Australia. PMID- 8401061 TI - Implementing clinical guidelines. Guidelines may not be cost effective. PMID- 8401062 TI - Implementing clinical guidelines. Computers allow instant access. PMID- 8401063 TI - Information overload. PMID- 8401064 TI - NHS information technology. PMID- 8401065 TI - Antenatal screening for Down's syndrome. Performance of different markers remains confused. PMID- 8401066 TI - Antenatal screening for Down's syndrome. Counselling may not help vulnerable women. PMID- 8401067 TI - Antenatal screening for Down's syndrome. The west of Scotland's screening programme. PMID- 8401068 TI - Anophthalmia and gaps in public health policy. PMID- 8401069 TI - Risk of HIV infection in homosexual men. PMID- 8401070 TI - Transmission of HIV in prison. PMID- 8401071 TI - Improving uptake of immunisation. Mobile children miss out. PMID- 8401072 TI - Improving uptake of immunisation. What about parents who say "No"? PMID- 8401073 TI - Screening boys for delayed growth. PMID- 8401074 TI - Management of septic shock. PMID- 8401075 TI - Teenagers, sex, and risk taking. PMID- 8401076 TI - Management of trauma. PMID- 8401077 TI - How much alcohol is sensible? PMID- 8401078 TI - Termination of pregnancy with mifepristone. PMID- 8401079 TI - The double contrast barium enema. PMID- 8401080 TI - Seeing is believing. PMID- 8401081 TI - Minor surgery in general practice. PMID- 8401082 TI - The new NHS. Very peculiar practice. PMID- 8401083 TI - Health care in South Africa. PMID- 8401084 TI - The new NHS. Research and development. PMID- 8401085 TI - Medicine and global survival. PMID- 8401086 TI - Retinopathy of prematurity. PMID- 8401087 TI - Deregulating emergency contraception. PMID- 8401088 TI - Educating general practitioners. PMID- 8401089 TI - Rituals in antenatal care--do we need them? PMID- 8401090 TI - The routine six week postnatal vaginal examination. PMID- 8401091 TI - Prevalence of neural tube defects in South Australia, 1966-91: effectiveness and impact of prenatal diagnosis. AB - OBJECTIVE: To determine trends in total prevalence of neural tube defects in South Australia during 1966-91, the impact of prenatal diagnosis on birth prevalence, and the effectiveness of prenatal screening for neural tube defects in 1986-91. DESIGN: All births and terminations of pregnancy affected by neural tube defects and information on prenatal screening were ascertained from multiple sources including the South Australian perinatal and abortion statistics collections, birth defects register, and state maternal serum alpha fetoprotein screening programme. SETTING: Southern Australia. SUBJECTS: All 1058 births and terminations of pregnancy affected by neural tube defects in 1966-91. MAIN OUTCOME MEASURES: Total prevalence and birth prevalence of individual and all neural tube defects. The proportion of screened cases detected prenatally. RESULTS: Total prevalence of neural tube defects during 1966-91 was 2.01/1000 births with no upward or downward trend. However, birth prevalence fell significantly (by 5.1% a year), with an 84% reduction from 2.29/1000 births in 1966 to 0.35/1000 in 1991 (relative risk = 0.16, 95% confidence interval 0.07 to 0.34). The fall was 96% for anencephaly and 82% for spina bifida. 85% of defects, both open and closed, were detected before 28 weeks' gestation in women screened by serum alpha fetoprotein or mid-trimester ultrasonography, or both, in 1986-91 (99.0% for anencephaly and 75.7% for spina bifida). CONCLUSIONS: While the total prevalence of neural tube defects in South Australia remained stable, prenatal diagnosis and termination of pregnancy resulted in an 84% fall in birth prevalence during 1966-91. Screening detected over four fifths of cases in 1986 91. PMID- 8401092 TI - Outcome in colorectal adenocarcinoma: two seven-year studies of a population. AB - OBJECTIVE: To record every patient with proved colorectal adenocarcinoma presenting from a defined population over two years in 1968-9 and during 1980-2, and to compare treatment and outcome over seven years. DESIGN: Retrospective in 1968-9 and prospective in 1980-2. SETTING: Aberdeen general hospitals. SUBJECTS: Every patient presenting to the four general surgical units with histological proof of colorectal adenocarcinoma. MAIN FINDINGS: On average, one new patient presented each week per 100,000 population. The proportion of patients aged 65 and over rose from 67% to 71%. An operation was performed on 385 patients in 1968 9 and on 399 during 1980-2. At laparotomy the proportion of patients who seemed to be curable and had a radical operation rose slightly, from 56% to 61%, and operative mortality fell from 9% to 5%. In all there were 421 survivors of curative surgery, and seven years later three quarters were either alive (51%) or had died without recurrence (25%). In both studies some 40% of patients were considered incurable when they presented, but the number who had a palliative resection rose from 59% to 85%. CONCLUSIONS: The contribution of radical surgery to the treatment of colorectal adenocarcinoma is substantial, with three quarters of patients so treated showing no evidence of recurrence after seven years. Operative mortality in an elderly population is now low, and improvement in late outcome is more likely to come from developments in adjuvant therapy than in operative technique. A continuing problem is that 40% of patients are not curable when they present, although palliative resection can now be offered to over 80%. PMID- 8401093 TI - Serum albumin concentration, arm circumference, and oedema and subsequent risk of dying in children in central Africa. AB - OBJECTIVE: To measure the prognostic value of clinical, anthropometric, and biological indicators of protein energy malnutrition in hospitalised children. DESIGN: Hospital based follow up study from admission to discharge or death of a cohort of children. SETTING-Paediatric hospital in Zaire. SUBJECTS: 1129 children consecutively admitted between August 1986 and October 1988. MAIN OUTCOME MEASURES: Height, weight, arm circumference, skinfold thicknesses, serum albumin concentration, and mortality. RESULTS: Mortality was higher in wasted children and in those with a mid-upper arm circumference < 125 mm, a serum albumin concentration < 16 g/l, and oedema. After multivariate analysis, serum albumin concentration was the best predictor of subsequent risk of dying. Mid-upper arm circumference and oedema, however, still contributed considerably to evaluation of mortality. CONCLUSIONS: In this specific environment of central Africa an isolated clinical sign such as oedema is not enough to detect children with a high risk of dying among those admitted to paediatric wards with severe protein energy malnutrition. Measurement of additional indicators such as arm circumference and serum albumin concentration seems to be of crucial importance. PMID- 8401094 TI - Comparison of medical abortion with surgical vacuum aspiration: women's preferences and acceptability of treatment. AB - OBJECTIVES: To assess women's preferences for, and the acceptability of, medical abortion and vacuum aspiration in the early first trimester. DESIGN: Patient centred, partially randomised trial. Medical abortion was performed with mifepristone 600 mg followed 48 hours later by gemeprost 1 mg vaginal pessary. Vacuum aspiration was performed under general anaesthesia. SETTING: Teaching hospital in Scotland. PATIENTS: 363 women undergoing legal induced abortion at less than nine weeks' gestation. MAIN OUTCOME MEASURES: Women's preferences for method of abortion before abortion; acceptability judged two weeks after abortion by recording the method women would opt to undergo in future and by semantic differential rating technique. RESULTS: 73 (20%) women preferred to undergo medical abortion, and 95 (26%) vacuum aspiration; 195 (54%) were willing to undergo either method, and were allocated at random. Both procedures were highly acceptable to women with preferences. Gestation had a definite effect on acceptability in randomised women; at less than 50 days there were no differences, but between 50 and 63 days vacuum aspiration was significantly more acceptable. CONCLUSIONS: Women who wish to use a particular method should be allowed their choice, regardless of gestation. Women of 50-63 days' gestation without preferences for a particular method are likely to find vacuum aspiration more acceptable. A patient centred, partially randomised trial design may be a useful tool in pragmatic research. PMID- 8401095 TI - Clinical features, risk factors, and referral delay in British patients of Indian and European origin with angina matched for age and extent of coronary atheroma. PMID- 8401096 TI - The academic base for general practice: the case for change. AB - University departments of general practice and the postgraduate education system for general practice have developed separately over the past 30 years. This separation is now impeding the academic development of the discipline and causes difficulties with recruitment and career progression. These problems could be eased by the creation of integrated departments. This would establish a critical mass for research and educational development, allow human and other resources to be used more flexibly and effectively, and provide a strong base for undergraduate education, vocational training, higher professional training, and continuing education. It could encourage collaborative ventures with other disciplines and also lead to higher standards of patient care. PMID- 8401097 TI - "Natural family planning": effective birth control supported by the Catholic Church. AB - During 20-22 September Manchester is to host the 1993 follow up to last year's "earth summit" in Rio de Janeiro. At that summit the threat posed by world overpopulation received considerable attention. Catholicism was perceived as opposed to birth control and therefore as a particular threat. This was based on the notion that the only method of birth control approved by the church--natural family planning--is unreliable, unacceptable, and ineffective. In the 20 years since E L Billings and colleagues first described the cervical mucus symptoms associated with ovulation natural family planning has incorporated these symptoms and advanced considerably. Ultrasonography shows that the symptoms identify ovulation precisely. According to the World Health Organisation, 93% of women everywhere can identify the symptoms, which distinguish adequately between the fertile and infertile phases of the menstrual cycle. Most pregnancies during trials of natural family planning occur after intercourse at times recognised by couples as fertile. Thus pregnancy rates have depended on the motivation of couples. Increasingly studies show that rates equivalent to those with other contraceptive methods are readily achieved in the developed and developing worlds. Indeed, a study of 19,843 poor women in India had a pregnancy rate approaching zero. Natural family planning is cheap, effective, without side effects, and may be particularly acceptable to the efficacious among people in areas of poverty. PMID- 8401098 TI - Costs and cost-minimisation analysis. AB - Whatever kind of economic evaluation you plan to undertake, the costs must be assessed. In health care these are first of all divided into costs borne by the NHS (like drugs), by patients and their families (like travel), and by the rest of society (like health education). Next the costs have to be valued in monetary terms; direct costs, like wages, pose little problem, but indirect costs (like time spent in hospital) have to have values imputed to them. And that is not all: costs must be further subdivided into average, marginal, and joint costs, which help decisions on how much of a service should be provided. Capital costs (investments in plant, buildings, and machinery) are also important, as are discounting and inflation. In this second article in the series Ray Robinson defines the types of costs, their measurement, and how they should be valued in monetary terms. PMID- 8401099 TI - Hepatitis B vaccination ... and given booster injections. PMID- 8401100 TI - Hepatitis B vaccination. Non-responders must be detected ... PMID- 8401101 TI - False reassurance of pulse oximetry. Take note of inspired oxygen concentration. PMID- 8401102 TI - False reassurance of pulse oximetry. Misunderstanding leads to dangerous practice. PMID- 8401103 TI - False reassurance of pulse oximetry. Use your nerve stimulator. PMID- 8401104 TI - Effect of aspirin and NSAIDs on colorectal adenomas. Protective effect may be spurious. PMID- 8401105 TI - Migraine and risk of stroke. PMID- 8401106 TI - Thyroid function in elderly people. Take account of iodine supply. PMID- 8401107 TI - Thyroid function in elderly people. Thyrotrophin releasing hormone test is useful. PMID- 8401108 TI - Sample size in audit. PMID- 8401109 TI - Generalized seizure due to terfenadine. PMID- 8401110 TI - Management of hyponatraemia. Differentiate between acute and chronic. PMID- 8401111 TI - Unexpected cardiac abnormalities in Lyme disease. PMID- 8401112 TI - Preventing melanoma. PMID- 8401113 TI - Managing impotence in diabetes. No need for separate service. PMID- 8401114 TI - Managing impotence in diabetes. Psychological factors remain important. PMID- 8401115 TI - Managing impotence in diabetes. Two drugs are better than one. PMID- 8401116 TI - Managing impotence in diabetes. Wait to be asked before offering advice. PMID- 8401117 TI - Clinton's health care reforms. PMID- 8401118 TI - Asthma: what is there left to find out? PMID- 8401119 TI - Optimal pain relief in infants and children. PMID- 8401120 TI - Clinical genetics meets primary care. PMID- 8401121 TI - Presenting expert evidence in criminal proceedings. PMID- 8401122 TI - Successful liver transplantation in babies under 1 year. AB - OBJECTIVE: To review the outcome of liver transplantation in babies aged less than 1 year. DESIGN: Prospective evaluation of survival, clinical complications, and nutritional and developmental status before and one year after liver transplantation. SETTING: The Children's Hospital and Queen Elizabeth Hospital, Birmingham. SUBJECTS: All 25 babies who received liver transplantation from January 1989 to December 1992 were included. Median age was 9 months and median weight was 7.0 kg. Seven babies were assessed but were not given transplants because they died while on the waiting list (two) or had severe extrahepatic disease (five). RESULTS: 24 babies had severe decompensated liver disease and 20 were severely malnourished despite nutritional support. Six babies received a whole liver graft and 19 received a reduction hepatectomy. Postoperative complications included primary nonfunction of the transplanted liver (one baby), hepatic artery thrombosis (two), biliary obstruction (seven), acute and chronic rejection (six), and sepsis (18). Three babies required a second transplant; all survived. Three babies, two of whom presented with fulminant hepatic failure, died. The overall actuarial survival rate (4 months to 4 years) is 88%. Review at 12 months showed a dramatic improvement in growth (p < 0.001) and normal psychosocial development with good quality of life. CONCLUSION: The improvement in survival rates and quality of life in this group of very sick babies is related not only to the development of reduction hepatectomy but also to advances in medical and nursing expertise. Early referral for liver transplantation is justified even if babies are critically ill. PMID- 8401123 TI - Radioiodine treatment of multinodular non-toxic goitre. AB - OBJECTIVE: To investigate the long term effect of radioactive iodine on thyroid function and size in patients with non-toxic multinodular goitre. DESIGN: Consecutive patients with multinodular non-toxic goitre selected for radioactive iodine treatment and followed for a minimum of 12 months (median 48 months) after an intended dose of 3.7 MBq/g thyroid tissue corrected to a 100% uptake of iodine 131 in 24 hours. PATIENTS: 69 patients with a growing multinodular non-toxic goitre causing local compression symptoms or cosmetic inconveniences. The treatment was chosen because of a high operative risk, previous thyroidectomy, or refusal to be operated on. MAIN OUTCOME MEASUREMENTS: Standard thyroid function variables and ultrasonically determined thyroid volume before treatment as well as 1, 2, 3, 6, and 12 months after treatment and then once a year. RESULTS: 56 patients were treated with a single dose of 131I, 12 with two doses, and one with four doses. In 45 patients treated with one dose and remaining euthyroid the median thyroid volume was reduced from 73 (interquartile range 50-106) ml to 29 (23-48) ml at 24 months in the 39 patients in whom this was measured during follow up. The median reduction was 40 (22-48) ml (60% reduction, p < 0.0001), half of which occurred within three months. Patients treated with two doses as well as those developing hypothyroidism and hyperthyroidism had a significant reduction in thyroid volume. Eleven patients developed hypothyroidism (cumulative five year risk 22%, 95% confidence interval 4.8% to 38.4%). Side effects were few: three cases of hyperthyroidism and two cases of radiation thyroiditis. Only one patient was dissatisfied with the result; she was referred for operation six months after treatment. CONCLUSIONS: A substantial reduction in thyroid volume accompanied by a low incidence of hypothyroidism and few side effects makes the use of radioactive iodine an attractive alternative to surgery in selected cases of non-toxic multinodular goitre. PMID- 8401124 TI - Association between impaired glucose tolerance and circulating concentration of Lp(a) lipoprotein in relation to coronary heart disease. AB - OBJECTIVE: To examine whether impaired glucose tolerance and raised Lp(a) lipoprotein concentrations are associated in subjects with coronary artery disease. DESIGN: Study of two subject populations, one with and one without symptomatic coronary artery disease. Case-control analysis of patients with impaired glucose tolerance and normal glucose tolerance performed in each subject population independently. SETTING: A general practice and a hospital ward in Newcastle upon Tyne. SUBJECTS: 517 apparently healthy subjects, 13 with impaired glucose tolerance, and 245 patients who had undergone coronary artery bypass graft surgery 12 months before, 51 with impaired glucose tolerance. MAIN OUTCOME MEASURES: Serum Lp(a) lipoprotein concentration, plasma glucose concentration before and after oral challenge with 75 g glucose monohydrate, and Lp(a) lipoprotein isoforms. RESULTS: In both the asymptomatic subjects and the subjects with coronary artery disease there was no significant difference between subjects with impaired glucose tolerance and subjects with normal and body mass index in serum Lp(a) lipoprotein concentrations (geometric mean 61 (geometric SD 4) mg/l v 83 (5) mg/l for asymptomatic subjects, 175 (3) v 197 (2) for subjects with heart disease), nor was there any difference in the proportion of subjects who had Lp(a) lipoprotein concentrations > 300 mg/l (31% v 23% for asymptomatic subjects, 37% v 37% for subjects with heart disease). For both subject groups there was no significant correlation between Lp(a) lipoprotein concentration and plasma glucose concentration after a glucose tolerance test, nor did Lp(a) lipoprotein concentration vary by quintile of glucose concentration after the test. Examination of Lp(a) lipoprotein isoforms in the subjects with coronary artery disease revealed an inverse relation between isoform size and plasma Lp(a) lipoprotein concentration, but there was no evidence that impaired glucose tolerance was associated with particular Lp(a) lipoprotein isoforms. CONCLUSION: Raised Lp(a) lipoprotein concentrations are not responsible for the association between impaired glucose tolerance and coronary artery disease. PMID- 8401125 TI - Measuring the impact of menopausal symptoms on quality of life. AB - OBJECTIVE: To examine the impact of menopausal symptoms on the overall quality of life of women. DESIGN: Data collection with a questionnaire administered by an interviewer, incorporating two different quality of life measurement techniques (time trade off and rating scale). SETTING: Specialist menopause clinic and two general practices in Oxford. SUBJECTS: 63 women aged 45-60 years recruited opportunistically during a clinic or appointment with a general practitioner; no exclusion criteria. RESULTS: Subjects gave very low quality of life ratings for health states with menopausal symptoms. The time trade off method of measuring preferences for these health states (on a scale from 0 to 1, where preference for full health is given as 1) yielded utility values of 0.64 for severe menopausal symptoms and 0.85 for mild symptoms. The rating scale measurement technique yielded even lower values: utilities of 0.30 and 0.65 were obtained for severe and mild symptoms respectively. Kappa scores indicated that the two methods produced results that were poorly related but not contradictory. Comparison of quality of life ratings before and after treatment with hormone replacement therapy showed significant improvements: with the rating scale measurement technique mean increases in utility values after the relief of severe and mild menopausal symptoms were 0.56 and 0.18 respectively. CONCLUSIONS: Quality of life may be severely compromised in women with menopausal symptoms, and perceived improvements in quality of life in users of hormone replacement therapy seem to be substantial. This emphasises the need to include quality of life measurements when assessing outcomes of hormone replacement therapy. Several limitations may exist with widely applied measurement techniques, calling for the development of appropriate and well validated instruments for measuring quality of life associated with reduced health states. PMID- 8401126 TI - Effects of time, sex, ethnic origin, and area of residence on prevalence of asthma in Israeli adolescents. AB - OBJECTIVES: To study effects of time, sex, ethnic origin, and area of residence on prevalence of asthma in Israeli adolescents. DESIGN: Retrospective survey of asthma from computerised medical draft records of conscripts examined up to the end of 1989. SETTING: Five regional centres in Israel. SUBJECTS: 443 186 conscripts (262 836 males and 180 350 females) aged 17-18 who were born over a nine year period. MAIN OUTCOME MEASURES: Asthma determined by medical history, physical examination, and lung function tests at rest and after exercise. RESULTS: Asthma was more prevalent in males than females (26.5/1000 v 21.4/1000, relative risk 1.25 (95% confidence interval 1.19 to 1.32)). Subjects were split into three groups according to year of birth, and prevalence of asthma increased over time from 18/1000 to 24/1000 to 36/1000 (risk of asthma 0.56 (0.54 to 0.59) for first birth group relative to last birth group and 0.69 (0.66 to 0.72) for second group relative to last group). Risk of asthma was also affected by ethnic origin (highest for Western origin and lowest for north African origin, relative risk 1.63 (1.56 to 1.71)) and area of residence (highest in the central coastal region--risk of 1.24 (1.19 to 1.30) relative to the north coastal region--and lowest in inland areas--risk of 0.67 (0.64 to 0.70) relative to north coastal region). CONCLUSIONS: Prevalence of asthma in Israel is increasing and is higher in males, in people of Western origin, and in those living in the most industrialised coastal region. PMID- 8401127 TI - Drivers who defy the law. PMID- 8401128 TI - Assaults on professional carers of elderly people. PMID- 8401129 TI - Primary non-compliance with prescribed medication in primary care. AB - OBJECTIVE: To determine the rate of patients not redeeming their prescriptions (primary noncompliance) and assess the factors influencing this. DESIGN: Observational study comparing copies of prescriptions written by general practitioners with those dispensed by pharmacists and subsequent case record review. SETTING: A large rural general practice in Tayside. SUBJECTS: All 4854 patients who received prescriptions (20,921) written between January 1989 and March 1989. MAIN OUTCOME MEASURES: The rate of non-redemption of prescriptions. RESULTS: Seven hundred and two patients (14.5%) did not redeem 1072 (5.2%) prescriptions during the study period, amounting to 11.5% of men and 16.3% of women. Non-redemption was highest in women aged 16-29 (27.6% of women) and men aged 40-49 (18.3% of men). Of prescriptions issued to women for oral contraceptives 24.8% were not redeemed during the study period. In those who redeemed prescriptions 17% were not exempt from prescription charges compared with 33% of patients who failed to redeem them. The non-redemption rate was highest for prescriptions issued at the weekends, although this was a small proportion of all prescribing. Prescriptions issued by trainee general practitioners were also less likely to be redeemed. CONCLUSIONS: Non-redemption varies with age, sex, general practitioner, exemption status, and with day of the week the prescription was written. Observational studies of drug exposure can be more accurately estimated from dispensing rather than prescribing data. PMID- 8401130 TI - Screening for carriers of cystic fibrosis--a general practitioner's perspective. AB - The identification of the gene for cystic fibrosis has led to the possibility of population based screening for carriers of cystic fibrosis to identify couples at risk of having an affected child. Pilot studies have shown that screening is feasible and does not cause untoward anxiety, though the uptake of testing varies considerably with the setting and method of invitation. Screening offered at times when individuals (and health professionals) perceive it as directly relevant will probably gradually become established in the United Kingdom. This review examines the role of general practice in genetic carrier screening as exemplified by cystic fibrosis. General practice has a pivotal role from the beginning in providing individuals and couples with information, facilitating testing of patients' relatives and of carriers identified by screening elsewhere (such as antenatal clinics), and offering testing in the context of reproduction. Screening for the cystic fibrosis gene will probably be followed by other genetic screening programmes. PMID- 8401131 TI - Adult moyamoya disease. PMID- 8401132 TI - ABC of emergency radiology. Cervical spine--II. PMID- 8401134 TI - Transferring patients with serious head injury. PMID- 8401133 TI - Cost-utility analysis. AB - Decisions have to be made about allocating health resources. Currently the best economic evaluation method for doing this is cost-utility analysis. This compares the costs of different procedures with their outcomes measured in "utility based" units--that is, units that relate to a person's level of wellbeing. The most commonly used unit is the quality adjusted life year (QALY). QALYs are calculated by estimating the total life years gained from a procedure and weighting each year to reflect the quality of life in that year. To compare outcomes of different programmes the Rosser index is one measure that is widely used to assign quality of life scores to patients. Combined with a measure of life years gained from a procedure, this enables QALYs to be calculated and procedures ranked according to cost per QALY gained. In this article Ray Robinson explains the measures used and discusses how QALY league tables can be used to guide decisions on resource allocation. PMID- 8401135 TI - Incidence of tuberculosis in England and Wales. Rate in ethnic groups is decreasing. PMID- 8401136 TI - Attempted suicide. Are effective disorders missed. PMID- 8401137 TI - Incidence of tuberculosis in England and Wales. Europeans may be more at risk. PMID- 8401138 TI - Serum screening for Down's syndrome. Not adequately validated. PMID- 8401139 TI - Managing neck injuries. PMID- 8401140 TI - Prioritising resources. PMID- 8401141 TI - Fatigue varies according to where it is measured. PMID- 8401142 TI - Incidence of breast cancer. PMID- 8401143 TI - Managing patients in a persistent vegetative state. PMID- 8401144 TI - Retinoblastoma and fetal irradiation. PMID- 8401145 TI - Junior surgeons lack practical experience. PMID- 8401146 TI - War in former Yugoslavia. Disabled children in the pink zone. PMID- 8401147 TI - War in Bosnia. Lessons from Vietnam. PMID- 8401148 TI - Skeletal muscle metabolic enzymes are altered by hyperbaric oxygenation treatments. AB - To test whether repeated HBO exposures would increase activity of skeletal muscle metabolic enzymes, 27 rabbits (3 groups) were exposed 90 min/day, 5 days/wk to either 100% O2 at 243 kPa (HBO), 100% O2 at 101 kPa (HIO), or 21% O2 at 101 kPa (CON). Four animals per group were killed after 2 wk treatment, and the remaining five per group were killed after 8 wk of treatment. Soleus, plantaris, and tibialis anterior muscles were removed, and the activities of adenylate kinase, alpha-glycerophosphate dehydrogenase, and citrate synthase were measured. After 8 wk there was no difference in enzyme activity between groups for either plantaris or tibialis anterior. In the soleus after 8 wk there was no difference between groups in adenylate kinase activity, but alpha-glycerophosphate dehydrogenase activity was 56% greater (P < 0.05) in HBO than in HIO and 50% greater than in CON, and citrate synthase activity in HBO was 24% greater (P < 0.05) than that in HIO and 36% greater than that in CON. Inasmuch as the soleus is a postural muscle, these results suggest that long-term HBO treatments can increase enzyme activity in an actively contracting muscle. PMID- 8401149 TI - Effect of a single exposure to hyperbaric oxygen on blood mononuclear cells in human subjects. AB - We studied the effect of a single exposure to a therapeutic profile of hyperbaric oxygen on blood mononuclear cell subset. Twenty healthy volunteers were exposed to 0.28 MPa for 90 min. Thirteen breathed pure oxygen and seven were control subjects exposed to compressed air at the same pressure. Venous blood samples were drawn before HBO exposure, immediately on exit from the chamber, and 24 h later. Immediately after the exposure, a significant increase was observed in the percentage and absolute number of CD8 (suppressor/cytotoxic) T cells, with a concomitant decrease in the CD4 (helper/inducer) T cells. These changes resulted in a decreased CD4:CD8 ratio. A rise was also observed in the number of HLA-DR antigen-bearing cells, with a transient increase in monocytes. There was no change in the total count and percentage of T cells (CD3), B cells, and NK cells. Twenty-four hours after HBO exposure there was a partial reversal of the decrease in the mean CD4:CD8 ratio, but it was still significantly lower than preexposure values. The fast reversibility of the change in the CD4:CD8 ratio suggests specific HBO-induced shifts and sequestration of T-cell subpopulations. PMID- 8401150 TI - Are beta-endorphins and thermoregulation during cold-water immersion related? AB - The purpose of this investigation was to determine the relationship between core temperature (Tre), tissue insulation (I), and beta-endorphins (beta-END) during immersion in cold water. To test this, 21 males were immersed to the first thoracic vertebra for 120 min in stirred water at either 18 degrees, 22 degrees, or 26 degrees C. Subjects were divided with respect to body fat [high fat (HF) = 18-22%, n = 11; low fat (LF) = 8-12%, n = 10] and randomly assigned to one of the three temperatures. Blood was drawn at 25 and 5 min (T-25 and T-5, respectively) before immersion, after 1 h of immersion, and 5 min before the completion of the second hour (T + 60 and T + 120, respectively). No significant relationship was found between delta beta-END and delta I or delta Tre. The relationship between the changes in thermoregulatory variables and delta beta-END did not differ between the HF and LF groups. Also, beta-END (adjusted for plasma volume) was lower (P < 0.05) in the HF vs. the LF group. beta-END (pg.ml-1) increased (P < 0.05) as time increased from T-25 to T + 60 and were not progressive with duration of immersion. Thus, beta-END did not increase in proportion to level of heat loss. PMID- 8401151 TI - Effects of prolonged CO2 inhalation on shivering thermogenesis during cold-water immersion. AB - We investigated the effect of prolonged hypercapnia on human thermoregulation during immersion of seven male subjects in a 15 degrees C water bath until their esophageal temperature dropped to 35 degrees C or until 1 h had elapsed. In the control trial, subjects inspired room air, whereas in the other trial the inhaled gas mixture was a 4% CO2:20% O2:76% N2 gas mixture. Oxygen uptake (VO2, liter.min 1), inspired minute ventilation (VI, liter.min-1), esophageal temperature (Tes, degree C), mean unweighted skin temperature (Tsk, degree C), mean heat flux (Q, W.m-2), and electromyographic (EMG, mV) activity of the trapezius muscle were recorded. VO2 and integrated EMG (IEMG) activity were used as the primary indicators of shivering thermogenesis. There was a tendency for elevated VO2, albeit not significant, in the CO2 trial compared to the air trial. We observed no significant differences in the IEMG between the air and CO2 trials. These results suggest that prolonged inhalation of a gas mixture containing 4% CO2 does not have a significant inhibitory effect on shivering thermogenesis and does not enhance the cooling rate of the body core. The absence of any shivering attenuation is most likely due to the small blood PCO2 increase incurred by inhalation of 4% CO2, compensation of hypercapnic-induced respiratory acidosis, and a strong thermal drive from core and peripheral regions. It is unlikely that elevated PICO2 levels contribute significantly to the etiology of hypothermia in divers. PMID- 8401152 TI - Moderate hypercapnia: cardiovascular function and nitrogen elimination. AB - Elevated carbon dioxide concentrations frequently encountered in diving operations may have cardiovascular effects. If so, changes in nitrogen loading and elimination may be induced. To study this possibility, whole body nitrogen elimination rates were determined using a rebreathing apparatus and gas chromatographic measurement of N2 in expired gas in six subjects as they breathed mixtures of 3 and 5% CO2 with 21% O2 and a balance of Ar for 125 min. No significant differences were observed among mean N2 yields, which were 815 ml (95% confidence interval +/- 51 ml), 831 ml (+/- 38 ml), and 845 ml (+/- 57 ml) for 0, 3, and 5% CO2 mixtures, respectively. Simultaneous measurements of heart rate showed a significant increase while breathing 5% CO2 as compared to 3 and 0% CO2. The increases in heart rate were not accompanied by any significant change in cardiac output, mean arterial pressure, or tissue perfusion. We conclude that at these levels of hypercarbia, tissue perfusion is not influenced enough to cause any changes in whole-body N2 elimination. PMID- 8401153 TI - Relationship between venous bubbles and hemodynamic responses after decompression in pigs. AB - We present a new pig model for studying relationships between venous gas bubbles and physiologic effects during and after decompression. Sixteen pigs were anesthetized to allow spontaneous breathing. Eight of them underwent a 30-min exposure to 5 bar (500 kPa) followed by a rapid decompression to 1 bar (2 bar/min); the remaining eight served as controls. The pigs were monitored for intravascular bubbles using a transesophageal echocardiographic transducer, and bubble count in the two-dimensional ultrasound image of the pulmonary artery was used as a measure of the number of venous gas bubbles. Effects on physiologic variables of the pulmonary and the systemic circulations were either measured or estimated. We detected venous bubbles in all pigs after decompression, but the interindividual variation was large. The time course of changes in the mean pulmonary artery pressure, in the pulmonary vascular resistance, in the arterial oxygen tension, and in the pulmonary shunt fraction followed the time course of the bubble count. In contrast, such a relationship to the number of venous gas bubbles was not found for the immediate increase in mean arterial pressure and for the changes in the other variables of the systemic circulation. We conclude that the number of venous gas bubbles, as evaluated by the bubble count in the ultrasound image of the pulmonary artery, is clearly related to changes in the variables of the pulmonary circulation in this pig model. PMID- 8401154 TI - Hyperbaric oxygen as prophylaxis or treatment for radiation myelitis. AB - This animal study was designed to investigate HBO as a treatment or prophylaxis for radiation myelitis. All animals received identical spinal cord radiation doses of 69 Gy in 10 daily fractions. Group I received no HBO; group II began HBO at the onset of signs of myelitis; group III received HBO with prophylactic intent beginning 6 wk after irradiation; and group IV received both modalities on the same day, but radiation always preceded HBO by at least 4 h. HBO consisted of 90 min oxygen at 2.4 atm abs for 20 daily treatments. Animals were objectively assessed for the loss of certain neurologic reflexes indicative of four levels of myelitis. Although all animals progressed to severe myelitis, group III animals had group-averaged levels of myelitis consistently less than control. The differences were statistically significant for several weeks. Group IV animals progressed to severe myelitis much more rapidly than any other group. Additional study is justified by this trial. Key questions to be answered include the optimal timing of HBO to produce a beneficial rather than detrimental effect. PMID- 8401155 TI - [The moral problem of suicide]. AB - There are 2 important philosophical traditions discussing the moral problem of suicide. One tradition rejects suicide as an immoral act that is against either an absolute or transcendent order, or against socio-political responsibilities, or against the duties one has to obey with respect to your own individual life. The other tradition argues that a specifically "qualified" suicide is a morally good privilege of human freedom and self-determination. Weighting the pros and cons the debate seems to end in a drawn game, perhaps with little advantages for the position against suicide. In any case, in the end it comes down to the very question whether a human being is still able to experience that it is good that he exists and that it is good to be engaged in this existence or not. PMID- 8401156 TI - [Brief definitions of medical ethics concepts. 28. Measuring quality of life in medicine (II)]. PMID- 8401157 TI - Transdermal measurement of alcohol consumption. PMID- 8401158 TI - The sixth Thomas James Okey Memorial Lecture. Vietnam veterans' rapid recovery from heroin addiction: a fluke or normal expectation? AB - Between 1972 and 1974, the outcomes of army enlisted men who had served in Vietnam during 1970-71 were evaluated and compared with that of a matched group. This paper reports the major findings of that study with respect to frequency of narcotic addiction in and after Vietnam, and the major risk factors for Vietnam addiction and later relapse. Extraordinary access to records facilitated drawing the sample, locating it, and verifying interview responses. The surprisingly low levels of readdiction and the rarity of addiction to narcotics alone as compared with poly-substance dependence are findings still not entirely incorporated into public and scientific views of heroin addiction. Some defenses against that incorporation are examined. PMID- 8401159 TI - HIV risks for women drug injectors: heroin and amphetamine users compared. AB - The incidence of HIV infection among women in Europe and the US is growing rapidly. Women who inject drugs are particularly vulnerable--they may acquire the infection through sharing injecting equipment and through sexual contact with an infected male. Opioids and stimulants are reputed to have different effects upon sexual activity and a sex life enhanced by drugs may increase the risk further. In the context of two larger studies of HIV-related risk behaviours among opioid and amphetamine users in the North West of England, the sexual behaviour and sharing of two groups of female injectors were compared, one whose primary use was heroin, the other amphetamine. Reported incidence of sharing was similar and high. Marked differences were observed in sexual behaviour, amphetamine injectors reporting greater interest in sex and greater frequency of intercourse. However, more of them perceived their personal risk to infection through unprotected sex as negligible. Over 80% in both groups had engaged in unprotected sex in the 6 months prior to interview. Women injectors tend to have injecting partners--more research is needed if health education strategies are to be devised that will protect them. PMID- 8401160 TI - Evaluation of syringe-exchange for HIV prevention among injecting drug users in rural and urban areas of Wales. AB - This paper considers the prevention of HIV in rural and urban areas among both opiate and non-opiate drug injectors. A 2-year study evaluated specialist and community based syringe-exchange provision in Wales. Numbers of clients and patterns of attendance at eight syringe-exchange schemes were monitored together with comparative cross-sectional studies of attenders (n = 152) and non-attenders (n = 176) from the population of drug injectors in 1990 and 1991. A total of 1171 clients made 7553 visits in the 2-year period, 110,000 syringes were issued and 80% of needles and syringes were returned. There were few demographic differences between attenders and non-attenders, but large and significant differences in HIV risk behaviour; only 9% of attenders had recently shared syringes in 1990 (10% in 1991) compared to 41% of non-attenders (39% in 1991). The catchment areas of specialised services were limited (5 miles or less) and insufficient in rural areas. Alternative community approaches to syringe distribution and exchange are examined. PMID- 8401161 TI - Factor analysis of ICD-10 symptom items in the 1988 National Health Interview Survey on Alcohol Dependence. AB - The present paper analyzes data from the 1988 National Health Interview Survey (NHIS88) of US adults. This general population survey is important in that it is the largest survey to date that collects detailed information on alcohol consumption and alcohol problems. The NHIS88 is a multi-stage complex sample household study with a sample size of about 44,000 adults of which about 20,000 reported that they were current drinkers. Symptom items were used for classification of individuals into categories of alcohol abuse and alcohol dependence based on the DSM-III-R, DSM-IV, and ICD-10 classification. The present analyses are based on 20 items corresponding to the ICD-10 classification, the elements of which are closely related to the Edwards-Gross alcohol dependence syndrome concepts. A single dimension corresponding to alcohol problems in general accounted for much of the correlations among these items, but additional dimensions of interest were also identified. Of the ICD-10 elements only Impaired Control, Tolerance, and Withdrawal could be identified as separate dimensions. PMID- 8401162 TI - The dimensionality of alcohol abuse and dependence: factor analysis of DSM-III-R and proposed DSM-IV criteria in the 1988 National Health Interview Survey. AB - Decisions on the final version of the DSM-IV alcohol abuse and dependence criteria will be determined largely by the APA's substance abuse field trials, conducted primarily in treated, clinical samples. Among the major objectives of the field trials are to study the boundaries between abuse and dependence, and to identify specific criteria that define the abuse category. The decisions on revisions of the abuse and dependence criteria in DSM-IV should, however, be informed by data from non-treated or general population samples as well. The present study addresses the field trial objectives using recent data from a large general population survey, the 1988 National Health Interview Survey (NHIS88). The paper reports on factor analyses to assess the dimensions underlying the DSM III-R and DSM-IV dependence and abuse criteria as operationalized in the NHIS88. The focus of the analyses is on whether models with more than one dimension are needed and if so, the correspondence of the dimensions to criteria sets defined in the DSM-III-R and DSM-IV. The analyses show that a two-dimensional model is required. The dimensions are interpreted as abuse and dependence, but the sets of criteria that define each of the dimensions show important deviations from the criteria sets used in the DSM definitions. PMID- 8401163 TI - Alcohol-related problems among Aboriginal drinkers in the Kimberley region of Western Australia. AB - The paper reports on the prevalence of alcohol-related problems among drinkers in a stratified random sample of the adult Aboriginal population of the Kimberley region of Western Australia. Subjects were 265 current drinkers who were identified in the total sample of 516 Kimberley Aboriginal men and women over the age of 15 years. Participants' reports were obtained on their frequency and quantity of alcohol consumption, and their lifetime experience of 16 alcohol related problems. The majority of Aboriginal drinkers in the Kimberley consumed harmful amounts of alcohol, and there was a high prevalence of the 16 alcohol related problems which showed a high degree of internal coherence, with the first principal component accounting for 45% of the total variance. The number of alcohol-related problems which respondents reported was strongly related to the quantity and frequency of self-reported alcohol consumption. PMID- 8401164 TI - Drinking contexts and drinking problems among black and white women. AB - This study explored whether black and white women differ in how often they drink in particular types of social settings and if drinking in different contexts independently predicts alcohol-related problems. The analysis was based on the interview responses of 635 black and 663 white women drinkers who represent sub samples from a nationwide survey of 5221 respondents conducted in 1984. The findings revealed that white women are more likely to attend restaurants, bars and parties away from home than black women and that a larger proportion of their alcohol consumption occurs in these settings than among black women. Factor analysis was used to develop scales on the the frequency of drinking in different social contexts. The results confirmed a three-dimensional factor structure that distinguished between drinking at home; drinking in social settings such as bars, restaurants and parties; and drinking in outdoor public areas like streetcorners and parks. A simultaneous equations path analysis was used to model the relationships among drinking contexts, the frequency of heavier drinking, drinking problems, race and other social characteristics. The major findings of the resulting models were that drinking contexts independently predict drinking problems and that race is not directly associated with drinking contexts or alcohol-related problems. However racial differences do exert significant indirect effects on social settings and drinking problems through differences in socio-economic status and normative attitudes. The conclusion emphasizes the complexity of the interrelationships of ethnic and social characteristics that underlie visible racial differences in the social patterns and situational contexts of alcohol use. PMID- 8401165 TI - The role of alcohol-related expectancies in the prediction of drinking behaviour in a simulated social interaction. AB - This study investigated the relationship between socially-relevant alcohol related beliefs and drinking behaviour in a laboratory-based study. Ninety male subjects were told they would be videotaped while completing the Simulated Social Interaction Test; they were then assigned to one of three conditions. In the first, alcohol consumption prior to the interaction was optional, in the second, a standard dose was administered, and in the third, no alcohol was offered. The outcome measures included drinking behaviour and rated level of social skill. In the optional condition, beliefs that alcohol would reduce anxiety in social situations and enhance social competence predicted amount consumed. The decision to continue drinking after the compulsory dose was predicted by anxiety level. In the no alcohol condition, rated social skill was negatively correlated with the beliefs that alcohol reduces tension and reduces fear of negative evaluation. Results are discussed with reference to social learning approaches to understanding drinking behaviour. PMID- 8401166 TI - Individual differences in differentiation among alcohol expectancy domains. AB - Prior research indicates that alcohol-related outcome expectancies represent important etiological factors in the understanding of alcohol use/abuse. Although current multidimensional measures assess several substantively different domains of alcohol-related outcome expectancies, there is growing evidence that they may not possess adequate levels of discriminant validity. Therefore, the present study sought to examine whether reliable between-person differences exist in the ability to differentiate among alcohol expectancy domains. The focus of the study was on three sets of intrapersonal characteristics: cognitive resources, cognitive constraints, and alcohol-related experience. Data were collected via household interviews with a random sample of 1125 adults. Multiple regression analysis revealed that higher levels of cognitive resources were associated with increasing levels of differentiation among alcohol expectancy domains. Results are discussed in terms of implications for the development of new or revised multidimensional alcohol expectancy questionnaires. Directions for future research are also discussed. PMID- 8401167 TI - Multivariate models for predicting abstention following intervention to stop smoking by general practitioners. AB - Predictors of successful smoking cessation were examined in a randomized controlled trial of 450 smokers who received an intervention by their general practitioner (GP). Pretreatment characteristics predicting outcome at 3, 6 and 12 months and for continuous abstinence to 12 months were determined using logistic regression analyses. Results showed the variables that significantly predicted abstention at 3 months were age and motivation, whereas the predictors at 6 months were socio-economic status, motivation, level of dependence and time spent with smokers. No single predictor emerged at 12 months. Predictors for continuous abstinence to 12 months were age, time spent with smokers and motivation. A model was developed which best describes the likelihood of patients achieving continuous long-term abstinence. The results show that five factors (high motivation level, older age, less time spent with smokers, low dependence level, and higher socio-economic status) together have a 76% accuracy of predicting continuous abstinence to 12 months. PMID- 8401168 TI - Prevalence of alcohol problems among selected AIDS risk groups: United States, 1988. AB - Estimates of the prevalence of various alcohol problems (DSM-III-R alcohol abuse/dependence, heavier drinking, and consuming 5 or more/9 or more drinks per occasion) were examined for a general population group of 29,155 persons age 18 years and older at self-reported risk for contracting AIDS by several definitions. All AIDS risk groups showed elevated risk for alcohol problems. Results are presented separately for males and females and for various demographic groups (age, marital status, family income, education, and race). PMID- 8401169 TI - Public attitudes to passive smoking in Europe. PMID- 8401170 TI - Life-long physical activity and cancer risk among Finnish female teachers. AB - This cohort study on Finnish female teachers of physical education (PE) and languages (L) was performed as an attempt to evaluate the importance of physical activity as a risk determinant of cancer, PE representing a physically active and L a less active subcohort. All the PE and L teachers graduating from 1920 onwards were obtained from four registers compiled from 1958-73 supplemented by an extension cohort drawn from the union membership register 1984-91 (in total 1,499 PE and 8,619 L teachers). Results of a questionnaire to representative samples of L and PE teachers did not reveal any major intergroup differences in social status, general health status, nutrition, maturation history, reproductive history, consumption of alcohol, smoking, or diet. However, the PE teachers reported higher life-long physical activity values than the L teachers. During the follow-up period 1967-91, the number of cancer cases totalled 108 for the PE and 513 for the L teachers. Expected numbers of cancer cases were calculated on the basis of national incidence figures, and the standardized incidence ratios (SIRs) were defined as ratios of observed and expected numbers of cases. The SIR for total cancers was 1.1 for the PE and 1.2 for the L teachers. In both teacher groups the SIRs for cancers of the breast, endometrium, ovary and colon were similarly elevated and the SIR for cervical cancer was reduced. There was an increased SIR for lung cancer (1.4) and skin melanoma (2.0) in the PE but not in the L teachers (0.5 and 0.9, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401171 TI - Anti-Helicobacter pylori antibodies prevalence and gastric adenocarcinoma in Portugal: report of a case-control study. AB - Evidence from large cohort studies has established an increased risk of gastric cancer for individuals infected with Helicobacter pylori (HP). In low incidence countries, like the United Kingdom and Sweden, case-control studies suggested that the prevalence of anti-HP antibodies in gastric cancer patients (at the time of cancer diagnosis) is greater than in control populations. We present results from a case-control study of the prevalence of IgG anti-HP antibodies in gastric cancer patients and a control population in a country with a high incidence of gastric cancer. Sera were studied from 80 gastric cancer patients (GC group) admitted consecutively to our department in 1990/91, and from 80 controls (CT group) matched by age and sex. IgG anti-HP was determined by ELISA. Patients' files were reviewed for evidence of previous diagnosis of peptic ulcer, gastric surgery, tumor localization and histopathological classification. Controls were submitted to a questionnaire for past history of peptic ulcer and gastric surgery. Positive results for anti-HP were: gastric cancer patients, 70.0%; control group, 81.5% (NS). However, the median optical densities (OD, a measure of antibody concentration) were significantly lower in the gastric cancer group than in controls: gastric cancer patients, 0.720 +/- 0.424 OD; control group, 0.906 +/- 0.443 OD (P = 0.004). There were no differences concerning past history of peptic ulcer or surgery. The proportion of positives for cancer of the cardia (66.7%) was lower than for the other tumour localizations (70.4%) (NS). Anti-HP positivity was lower in patients with gastric cancer associated with intestinal metaplasia than in controls (P = 0.14).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401172 TI - Instability of N-nitroso compounds in gastric juice and preliminary results from analyses of fresh samples by using an improved analytical method. AB - Using an improved method for determination of total N-nitroso compounds (NOC), we examined the stability of those compounds in fresh gastric juice samples during storage and the effects of the addition of 2% sulphamic acid on NOC concentration in 212 samples. The NOC levels in fresh samples decreased very rapidly at -20 degrees C, especially during the first 24 hours of storage (P < 0.01), and nitrite concentration also showed a decreasing trend during storage. The addition of sulphamic acid significantly reduced NOC levels from 1.97 +/- 0.21 to 1.10 +/- 0.12 mumol/l (mean +/- SE, P < 0.01), especially in samples of initially high pH. However, in some individual samples (16.5%) the NOC levels actually increased by 14.1% (P < 0.01). The results from analysis of NOC in 212 fresh samples in relation to pH demonstrated two significantly higher peaks of NOC concentrations at intragastric pH ranges 1.1 to 2.99 (P < 0.05) and 6.0 to 7.9 (P < 0.01). There was a significant relationship between nitrite level and intragastric pH (r = 0.480, P < 0.01), the nitrite concentration increasing dramatically when the pH exceeded 6.0. The present study suggest that a major proportion of the unidentified NOC formed through intragastric nitrosation is labile NOC; if the true concentration of NOC is to be determined it is therefore essential to analyse fresh gastric juice samples directly after collection and without pretreatment. It will also be necessary to characterize those labile NOC in order to study further the mechanism of endogenous N-nitrosation in man and its relation to human carcinogenesis. PMID- 8401173 TI - Cell kinetics of the in vitro metaphase arrest technique and the clinical applications. AB - The in vitro metaphase arrest technique (crypt cell production rate-CPPR) has been used to measure human rectal mucosal proliferation. Study of preincubation times, dose response curves and lag phases suggest that a concentration of vincristine of 5 micrograms/ml and 16 hour preincubation with time point increments between 25 and 125 minutes give optimal conditions for measuring rectal mucosal proliferation. Twenty individuals had rectal CCPR repeated without intervention of any kind. Close correlation was found between the two values (r = 0.89 and P = 0.0001). The effect of polyethylene glycol bowel preparation was also studied in 35 subjects. There was good correlation (r = 0.66, P = 0.007). There was close correlation between rectal and caecal CCPR as measured in 20 patients who had colonoscopy (r = 0.72, P = 0.0003). The in vitro metaphase arrest technique is a useful parameter of rectal mucosal proliferation and may be used with confidence in a number of different clinical situations. PMID- 8401174 TI - Bile acid inhibition of xenobiotic-metabolizing enzymes is a factor in the mechanism of colon carcinogenesis: tests of aspects of the concept with glucuronosyltransferase. AB - A factor in colon carcinogenesis might be the partial defeat in colon epithelial cells of the protective enzymic barrier against xenobiotics, via bile acid inhibition of enzymes that detoxify mutagens. The applicability of aspects of this concept to glucuronosyltransferase, a phenol detoxification enzyme, was tested in a colon cancer cell line. Inhibition of glucuronidation of the test substrate, 4-methylumbelliferone, occurred at bile acid concentrations found in faecal water, and depended on pH for some bile acids. Lithocholate was the most inhibitory: the concentration causing 50% inhibition of the initial rate of glucuronidation (IC50) was about 3 microM at pH 7.4 and at pH 6.2. The inhibitory potency of deoxycholate and chenodeoxycholate increased when pH decreased, but still remained less than that of lithocholate: the IC50 for deoxycholate was 88.5 microM at pH 7.4, and 14.8 microM at pH 6.2, and for chenodeoxycholate the IC50 was 67.4 microM at pH 7.4, and 21.7 microM at pH 6.2. Cholate did not cause appreciable inhibition. The inhibitory effects were additive when lithocholate was present together with either deoxycholate or chenodeoxycholate. The results provide a mechanism for the comutagenicity of bile acids, a feature of which is the inter-relation of bile acid comutagenicity specifically with mutagens that are inactivated by a bile acid-inhibitable enzyme. The results are also in accord with the view that high concentrations of bile acids in solution in faecal water, especially lithocholate, are a risk factor for colon cancer. PMID- 8401175 TI - Alcohol and cancer of the colon and rectum. AB - The association between alcohol intake and colorectal cancer was examined in a population-based case-control study performed in Stockholm in 1986-88. The study included 352 cases of colon cancer, 217 cases of rectal cancer, and 512 controls. Relative risks, with 95% confidence intervals, were calculated for total alcohol intake and for different alcoholic beverages. Total alcohol intake (> or = 30 g 100% ethanol per day) was not associated with colon cancer (relative risk = 0.9, confidence intervals = 0.4-1.8) or rectal cancer (1.0, 0.4-2.1). There was no evidence supporting beverage specificity (for colorectal cancer and > or = 10 g 100% ethanol per day: beer 1.1, 0.6-2.0, wine 1.0, 0.4-2.7, spirits 1.0, 0.6 1.6). The associations did not vary according to gender or site within the large bowel. These analyses were adjusted for year of birth and gender (when appropriate). Further adjustments for diet, body mass or physical activity had little or no influence on the results. The present study does not support the hypothesis that alcohol plays an important role in the aetiology of cancer of the colon and rectum in a population with a relatively low alcohol intake. PMID- 8401176 TI - Calcium phosphate: an alternative calcium compound for dietary prevention of colon cancer? A study on intestinal and faecal parameters in healthy volunteers. AB - In an effort to reduce the risk of colorectal cancer development, oral calcium carbonate supplementation has been used in previous studies for the precipitation of cytotoxic bile acids and fatty acids. In human intervention trials its effect on mucosal hyperproliferation in the colorectum has not always been satisfactory. Because the complexation of calcium and bile acids requires the formation of calcium phosphate, we performed an intervention study in 14 healthy volunteers, giving them 1,500 mg calcium as Ca3(PO4)2 for 1 week. The effects of tricalcium phosphate on luminal and faecal parameters of cytolytic activity were evaluated before, during, and after calcium phosphate supplementation. The cytolytic activity of faecal water and intestinal alkaline phosphatase activity in faecal water were not affected by supplemental calcium phosphate. In duodenal bile, the proportion of cholic acid tended to increase, whereas that of chenodeoxycholic acid tended to decrease during calcium phosphate supplementation. Neither concentrations of total and individual faecal bile acids, nor that of faecal fat were affected during calcium phosphate supplementation. It is suggested that, although phosphate is involved in bile acid precipitation, phosphate competes for calcium in the binding of fatty acids. This might possibly explain the unchanged cytolytic potency of faecal water, and therefore does not make tricalcium phosphate a suitable calcium compound for dietary intervention. PMID- 8401177 TI - Role of interleukin-1 inhibitory molecules in therapy of acute and chronic myelogenous leukemia. AB - The poor outcome of conventional therapy of acute and chronic myelogenous leukemias (AML and CML) has prompted several groups to investigate new therapeutic directions. Data from various laboratories, including our own, indicate that both normal and leukemia precursors proliferate in response to growth factors. Furthermore, it has been shown that AML blasts, low-density cells from CML patients with advanced disease, and cultured bone marrow-adherent layers from CML blast crisis patients produce interleukin 1 (IL-1); this molecule may play a pivotal role in driving leukemia cell proliferation through autocrine or paracrine pathways. We have therefore hypothesized that interruption of the IL-1 mediated growth-stimulatory mechanism may suppress leukemia precursor multiplication. In searching for IL-1-inhibitory molecules that may be used clinically, we have investigated the in vitro effects of various IL-1 inhibitors including IL-1 receptor antagonist, soluble IL-1 receptors, and interleukin 4. Our studies suggest that IL-1 inhibitors can suppress clonogenic growth of cultured AML and CML progenitors and may hence be exploitable in clinical trials. PMID- 8401178 TI - Recurrence of Philadelphia chromosome-positive leukemia in donor cells after bone marrow transplantation for chronic granulocytic leukemia. AB - Allogeneic bone marrow transplantation has been shown to be a very effective therapy for Chronic Granulocytic Leukemia with long term disease free survivals in excess of 60%. Relapse rates remain low at 15% following histocompatible sibling transplants and lower rates following matched unrelated donor grafts. Relapse rates however, are higher if BMT is carried out in transformation or blast crisis. Leukemic relapse in donor cells following transplantation for CGL is a rare event. The occurrence of donor leukemia however, may be under reported as accurate and sensitive investigation of the origin of relapsed leukemia following BMT requires DNA based technologies. A possible mechanism of donor leukemia in CGL is transfection of donor cells with the chimeric gene which is unique to this disease. It is possible that the malignant cells found in transformed or blast crisis of CGL may have a greater potential to transfect donor haematopoietic material. Careful evaluation of the incidence of donor leukemia using molecular biology methods may elucidate the frequency of this event following BMT for CGL. PMID- 8401179 TI - Graft-versus-leukemia reactions in clinical bone marrow transplantation. AB - Immunologic reactions occurring after allogeneic bone marrow transplantation, known as graft-versus-leukemia effect, contribute significantly to the control of minimal residual disease and decreased risk of relapse post-transplant. Graft versus-leukemia reactions occur with or without graft-versus-host disease and are either mediated through T-cells or are independent of T-cells. The contribution made by graft-versus-leukemia reactions to improvement in overall survival depends upon the underlying type of leukemia, the nature of the marrow graft and the morbidity of any associated graft-versus-host disease. This article reviews the evidence demonstrating graft-versus-leukemia effect in clinical bone marrow transplantation, some approaches to dissociate graft-versus-leukemia from graft versus-host disease, and attempts to induce graft-versus-leukemia reactions in the setting of autologous bone marrow transplantation by immunotherapy. PMID- 8401180 TI - Combination of t(14;18) and a Burkitt's type translocation in B-cell malignancies. AB - The combination of chromosomal translocations associated with bcl-2 rearrangement [t(14;18)] and c-myc rearrangement [t(8;14), t(8;22), or t(2;8)] has infrequently been detected in lymphoproliferative disorders. We have recently identified four cases of a B-cell malignancy exhibiting this dual translocation. In addition to t(14;18), one case had t(8;14) and three had the t(8;22). One case presented as de novo acute lymphoblastic leukemia (ALL-L2), two as de novo high grade lymphomas and the fourth evolved to a "blastic" phase from a previously documented follicular lymphoma. Immunophenotyping and molecular analysis was performed on three of the cases: all were negative for terminal deoxynucleotidyl transferase (TdT) but were CD10 positive. Two of the three cases with t(8;22) were negative for surface immunoglobulin (SIg) and positive for HLA-DR. Rearrangement of the oncogene bcl-2 was identified in a single case by polymerase chain reaction (PCR) only. Similar to cases reported in the literature, all patients had a poor clinical outcome despite aggressive therapy. Dual translocation lymphoid malignancy has a relatively characteristic morphology and the diagnosis should be considered when there is a history of an antecedent low grade lymphoma or when there is discordance between the "blastic" morphology and the immunophenotype (TdT- and/or SIg+). Confirmation requires demonstration of the characteristic translocations. Recognition of this entity has significant clinical implications that may require consideration of alternate treatment strategies. PMID- 8401181 TI - Potential sites of infection that develop in febrile neutropenic patients. AB - Three hundred episodes of neutropenia were reviewed for the occurrence of potential sites of infection. Ninety sites (30 per cent) were identified at the onset of fever independent of initial bacteraemia which was encountered in 104 episodes (35%) and predominantly involved Gram-positive cocci. A further 90 sites were recorded involving mainly the lower respiratory tract (58%) and skin and soft tissue (18%). These changes evolved significantly later (mean of 5.1 and 4.3 days respectively) than did other foci which mainly presented at the onset of fever (p < 0.01). However the infectious aetiology was established in only 54 cases overall with fungi being responsible for 25 of 45 cases of lower respiratory tract infections with a known microbiological aetiology. The mortality associated with initial bacteraemia, focus at onset and unexplained fever was 11-14% while that associated with the development of a subsequent focus was 28% with lung infiltrates carrying the worst prognosis. Therefore rather than being seen as a final solution for possible infectious complications, empiric therapy provides an opportunity for daily review of the patient thereby increasing the likelihood of both explaining initial fever and diagnosing subsequent infection. PMID- 8401182 TI - Correlation of marrow iron patterns with disease status of chronic myelogenous leukemia. AB - Greater than 80% of patients in the stable phase of chronic myelogenous leukemia (CML) have no detectable stainable iron in the marrow yet have normal serum iron, total iron binding capacity, and serum ferritin values. We studied the pattern of marrow iron in 187 marrow aspirates from 67 patients with Philadelphia chromosome positive CML in the chronic, accelerated and blast crisis stages. Sequential marrow aspirates in 30 patients confirm that the switch from negative iron to positive iron is coincident with the development of the accelerated phase of blast crisis of CML. Our data suggest that the pattern of marrow iron staining in CML is a useful prognostic indicator of the disease evolution. PMID- 8401183 TI - Effect of recombinant human manganese superoxide dismutase on radiosensitivity of murine B cell leukemia (BCL1) cells. AB - Recombinant human manganese superoxide dismutase (SOD) protects cells from oxidative damage and is known to ameliorate post-irradiation damage in mice exposed to whole body or localized chest irradiation. The concept behind the present experiments was to investigate whether it is possible to improve the outcome in leukemia following total body irradiation used as part of the conditioning prior to allogeneic bone marrow transplantation. We determined whether SOD protects leukemic cells from the effects of ionizing irradiation both in vitro and in vivo. Murine B cell leukemia (BCL1) cells, derived from tumor bearing mice, were irradiated in vitro with or without SOD and injected into BALB/c mice. All mice receiving 10(4) unirradiated BLC1 cells developed leukemia and died within 19-39 days. In vitro exposure of BCL1 cells to 800 cGy or 1600 cGy abolished the potential to induce leukemia by inoculation with 10(4) or 10(6) BCL1 cells, respectively. Addition of SOD in vitro during irradiation increased the resistance of BCL1 cells to ionizing irradiation; all mice receiving 10(6) BCL1 cells previously exposed in vitro to 1200 cGy in the presence of SOD died of leukemia, whereas only 40% of mice receiving a similar inoculum of irradiated BCL1 cells died of leukemia. In contrast, when BCL1-bearing mice were irradiated with 600-800 cGy with or without intravenous injection of SOD (100 mg/kg) 30 minutes prior to irradiation, development of leukemia was unaffected. Residual leukemia cells following therapy were assessed by adoptive transfer of 10(5) spleen cells to secondary BALB/c recipients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401184 TI - Electron microscopic identification of parvovirus virions in erythroid and granulocytic-line cells in a patient with human parvovirus B19 induced pancytopenia. AB - In this short report we describe a patient with human parvovirus B19 (HPV B19) induced transient pancytopenia. Parvovirus virions were seen by electron microscopy in both erythroid and granulocytic precursors. Erythroid cells are not the only targets in these cases. We draw attention to this disorder so that physicians involved with hematological disorders and transplantation be more aware of this infection. PMID- 8401185 TI - Isolated peripheral nerve relapse masquerading as Guillain-Barre syndrome in a patient with acute lymphoblastic leukemia. AB - We report a 30 year old patient with acute lymphoblastic leukemia (ALL) whose leukemic relapse presented as an isolated symmetrical peripheral neuropathy with facial diplegia. Initially, this was consistent with a Guillain-Barre syndrome but the peripheral nerve biopsy revealed leukaemic infiltration. This was followed by a systemic relapse. Reports of peripheral nerve infiltration are scarce and to the best of our knowledge this is the first documented case of peripheral polyneuropathy as a presenting manifestation of ALL relapse. PMID- 8401186 TI - Extensive cutaneous plasmacytomas. AB - An unusual case of extensive cutaneous and subcutaneous plasmacytomas is reported. Breast involvement was also a feature in this individual. The relevant literature relating to this unusual presentation of myeloma is reviewed. PMID- 8401187 TI - Cytoplasmic GpIIb-IIIa and cytokine secretion by blasts in a case of megakaryoblastic transformation of essential thrombocythemia. AB - A 49-year-old woman with a four year history of therapy resistant essential thrombocythemia, progressed to acute leukemia that also proved refractory to chemotherapy. Blast cell features including immunophenotype, cytogenetics and in vitro cell cultures, suggested megakaryoblastic leukemia. In serum-free culture, blasts released GM-CSF and IL-6 which sustained autocrine growth and promoted normal myeloid and megakaryocytic colony formation. PMID- 8401188 TI - Chylous effusion complicating chronic lymphocytic leukemia. AB - We present a case of chylous effusion (CE) occurring in a patient with chronic lymphocytic leukemia (CLL), an observation which has rarely been reported. Therefore, CLL should be added to the differential diagnosis of nontraumatic chylothorax. CE in CLL can be successfully managed by irradiation of the mediastinum. PMID- 8401189 TI - Research in nutrition. PMID- 8401190 TI - The Consumer Protection Act revisited. PMID- 8401191 TI - Human immunodeficiency virus-1 infection in spouses of seropositive individuals. AB - BACKGROUND: Unprotected sex can lead to transmission of the human immunodeficiency virus-1 (HIV-1) to the spouse of an infected individual. We studied the incidence of HIV-1 infection in the spouses of cases diagnosed to have HIV-1 infection by serology and the polymerase chain reaction (PCR). METHODS: Blood samples collected from 9 index cases and their respective spouses were tested for HIV-1 infection by ELISA, Western blot (WB) and PCR as well as from 10 healthy individuals with no high-risk behaviour. DNA extracted from both plasma and peripheral blood mononuclear cells was amplified by PCR, using multiple primer pairs for distinct regions of the HIV-1 genome. Specificity of the PCR product was demonstrated by hybridization to an oligonucleotide probe. RESULTS: All the index cases which were seropositive by ELISA and WB were also positive by PCR of plasma extracted DNA. Eight of the spouses were seronegative. Of these seven were positive by PCR--one spouse was negative by ELISA but showed a p55 band on WB and was positive by PCR. One spouse was negative by serology and PCR. The spouse belonging to the lone concordant couple was positive by serology and PCR. Except for one index case, PCR signals were obtainable only from DNA extracted from plasma but not from the DNA extracted from peripheral blood mononuclear cells. The control samples were negative by serology and PCR. CONCLUSION: It is possible to detect HIV-1 infection by PCR using DNA extracted from plasma even when the individuals are negative by ELISA and WB. It can help in the early counselling of HIV infected persons and their spouses. PMID- 8401192 TI - Cost analysis of a primary health centre in northern India. AB - BACKGROUND: Cost data are useful in health planning, budgeting and for assessing the efficiency of services. However, such data are not easily available from developing countries. We therefore estimated the cost incurred for the year 1991 92 on a primary health centre in northern India, which is affiliated to an academic institution. METHODS: The total costs incurred included the capital costs for land, building, furniture, vehicles and equipment as well as the recurrent costs for salaries, drugs and vaccines, diesel and maintenance. Except for land, where the 'opportunity cost' was calculated, the current market rates were considered for all other factors. A discount rate of 10% was used in the study. RESULTS: A total of Rs 777,015 (US $24,282) was incurred on the primary health centre in the study year, 80% being recurrent costs. Salaries constituted 62% of the total costs. A sum of Rs 30 (US $0.94) per head per year on primary health care was being incurred. CONCLUSION: Salaries constitute the bulk of the cost incurred on health. Approximately Rs 28 (40%) of the Rs 69 spent per head per year on health services by the Government of India is incurred on providing primary health care services. PMID- 8401193 TI - Toxic peripheral neuropathies in the tropics. PMID- 8401194 TI - Morphometry of normal human colorectal epithelium. PMID- 8401195 TI - Treatment of aplastic anaemia: the role of androgens. PMID- 8401196 TI - A case of do or die? PMID- 8401197 TI - Management of dysfunctional uterine bleeding. AB - DUB is a common symptom which is mainly due to disorders of ovulation and local endometrial dysfunction. The diagnosis should be made only after excluding organic pelvic disease. Anaemia and other endocrine and systemic causes of DUB must be treated promptly. Spontaneous cure is also possible. However, in patients in whom the symptoms persist, drug treatment with combined oestrogen-progestogen, anti-prostaglandin and anti-fibrinolytic agents is often effective. Hysterectomy and hysteroscopic endometrial ablation are contemplated only in persistent DUB in older and parous women and radiotherapy has no place amongst the modern modalities of management. PMID- 8401198 TI - Training statisticians for biostatistical consultation. PMID- 8401199 TI - Eminent Indians in medicine. P. N. Chhuttani. PMID- 8401201 TI - Unconventional medicine--a new age frontier. PMID- 8401200 TI - All India Institute of Medical Sciences, New Delhi. PMID- 8401202 TI - The Consumer Protection Act and I. PMID- 8401203 TI - Attitudes to liver transplantation in India. PMID- 8401204 TI - Biochemistry--an undergraduate major whose time has come. PMID- 8401205 TI - Structure, function, and regulation of adrenergic receptors. AB - Adrenergic receptors for adrenaline and noradrenaline belong to the large multigenic family of receptors coupled to GTP-binding proteins. Three pharmacologic types have been identified: alpha 1-, alpha 2-, and beta-adrenergic receptors. Each of these has three subtypes, characterized by both structural and functional differences. The alpha 2 and beta receptors are coupled negatively and positively, respectively, to adenylyl cyclase via Gi or Gs regulatory proteins, and the alpha 1 receptors modulate phospholipase C via the Go protein. Subtype expression is regulated at the level of the gene, the mRNA, and the protein through various transcriptional and postsynthetic mechanisms. Adrenergic receptors constitute, after rhodopsin, one of the best studied models for the other receptors coupled to G proteins that are likely to display similar structural and functional properties. PMID- 8401206 TI - Synthetic chimeras of mouse growth factor-associated glandular kallikreins. I. Kinetic properties. AB - A series of six chimeric proteins, composed of fragments corresponding to either one or the other of the growth factor-associated mouse glandular kallikreins epidermal growth factor binding protein (EGF-BP) and the gamma-subunit of nerve growth factor (gamma-NGF)--were expressed in Escherichia coli and isolated, and their kinetic properties were characterized. The assembly of these synthetic proteases involved the substitution of regions of the proteins containing four specific surface loops that have been postulated to influence both kinetic specificity and the formation of growth factor complexes. The substrates utilized in the kinetic characterization of these chimeric kallikreins were tripeptide nitroanilides representing carboxyl termini of both the EGF and beta-NGF mature hormones, putative processing sites for these kallikreins in the precursors. Characterization of these hybrid enzymes demonstrates that Km and kcat kinetic constants may be independently affected by the regions utilized in construction of these chimeric kallikreins. Specifically, loop 1, located in the amino terminal region (Bode, W., et al., J. Mol. Biol. 164, 237-282, 1983), in gamma NGF enhanced the kcat for substrates containing threonine in the P2 position, as is the case during the processing of the carboxy terminus of the beta-NGF precursor. Also, the central regions of the kallikreins containing loop 2 and the kallikrein loop dictated the generally inverted Km and kcat kinetic constants observed between EGF-BP and gamma-NGF. Finally, in gamma-NGF the autolysis loop, found in the carboxyl terminal region, functions to lower the Km kinetic constant for a variety of substrates. The results allow previously characterized kinetic differences between EGF-BP and gamma-NGF to be interpreted in terms of specific regions of the proteins and identify a subset of amino acid positions responsible for these functional characteristics. PMID- 8401207 TI - Synthetic chimeras of mouse growth factor-associated glandular kallikreins. II. Growth factor binding properties. AB - Six chimeric constructs of the sequentially similar growth factor-associated kallikreins-epidermal growth factor binding protein (EGF-BP) and the gamma subunit of nerve growth factor (gamma-NGF)--have been expressed, and their ability to generate complexes with epidermal growth factor (EGF) and beta-NGF, analogous to the high molecular weight forms (7S NGF and HMW-EGF) found in the mouse submaxillary gland, evaluated. The chimeras are distinguished by the interchange of three regions composing the amino, middle, and carboxyl terminal regions that encompass four surface loops possibly involved in specific growth factor interactions. Native beta-NGF (along with native alpha-NGF) formed complexes indistinguishable from naturally occurring 7S NGF, characterized by an alpha 2 beta gamma 2 structure (where beta-NGF is itself a dimer), with recombinant (r) gamma-NGF and with a chimera in which the amino terminal region from EGF-BP was substituted. Two other chimeras containing either the middle or carboxyl terminal regions of gamma-NGF showed weaker ability to form 7S complexes. Thus, all chimeras containing two segments from gamma-NGF retained at least some ability to form the 7S complex. rEGF-BP reacted weakly with EGF, but the chimera composed of the amino and middle segments of EGF-BP and the carboxyl terminal segment of gamma-NGF formed a nativelike HMW-EGF complex. None of the other chimeras appeared to bind EGF. These results identify amino acid positions within each kallikrein that participate in strong growth factor interactions and demonstrate that, outside of active site contacts, different regions of the kallikreins are involved in the binding of EGF and beta-NGF, respectively. PMID- 8401208 TI - Prediction of the three-dimensional structures of the nerve growth factor and epidermal growth factor binding proteins (kallikreins) and an hypothetical structure of the high molecular weight complex of epidermal growth factor with its binding protein. AB - We have predicted the three-dimensional structures of the serine protease subunits (gamma-NGF, alpha-NGF, and EGF-BP) of the high molecular weight complexes of nerve growth factor (7S NGF) and epidermal growth factor (HMW-EGF) from the mouse submandibular gland (from the X-ray crystal structures of two related glandular kallikreins). The conformations of three of the six loops surrounding the active site are relatively well defined in the models of gamma NGF and EGF-BP, but three other loops are likely to have flexible conformations. Although the amino acid sequence of alpha-NGF is closely related to those of gamma-NGF and EGF-BP, it is catalytically inactive. Model-building studies on alpha-NGF suggested that mutations (in alpha-NGF) just prior to the active site serine (195) and an unusual N-terminal sequence are consistent with alpha-NGF having a zymogen-like conformation (similar to that in chymotrypsinogen). An hypothetical model of the quaternary structure of HMW-EGF has been constructed using this model of EGF-BP and the NMR structure of murine EGF. The C-terminal arm of EGF was modeled into the active site of EGF-BP based on data indicating that the C-terminal arginine of EGF occupies the S1 subsite of EGF-BP. Data suggesting one of the surface loops of EGF-BP is buried in the HMW-EGF complex and symmetry constraints were important in deriving a schematic model. A molecular docking program was used to fit EGF to EGF-BP. PMID- 8401209 TI - Determining protein loop conformation using scaling-relaxation techniques. AB - We recently developed a rapid loop closure algorithm in which bond lengths are scaled to constrain the ends of a segment to match a known distance and then gradually relaxed to their standard values, with boundary constraints maintained. Although the algorithm predicted the Zif286 zinc-finger loop to within approximately 2 A, it had a serious limitation that made its more general use tentative: it omitted the atomic environment of the loop. Here we report an extension of the algorithm to take into account the protein environment surrounding a given loop from the outset of the conformational search and show that it predicts structure with an efficiency and accuracy that could not be achieved without continuous environmental inclusion. The algorithm should be widely applicable to structure determination when complete experimental information is unavailable. PMID- 8401210 TI - Design of synthetic gene libraries encoding random sequence proteins with desired ensemble characteristics. AB - Libraries of random sequence polypeptides are useful as sources of unevolved proteins, novel ligands, and potential lead compounds for the development of vaccines and therapeutics. The expression of small random peptides has been achieved previously using DNA synthesized with equimolar mixtures of nucleotides. For many potential uses of random polypeptide libraries, concerns such as avoiding termination codons and matching target amino acid compositions make more complex designs necessary. In this study, three mixtures of nucleotides, corresponding to the three positions in the codon, were designed such that semirandom DNA synthesized by repeated cycles of the three mixtures created an open reading frame encoding random sequence polypeptides with desired ensemble characteristics. Two methods were used to design the nucleotide mixtures: the manual use of a spreadsheet and a refining grid search algorithm. Using design targets of less than or equal to 1% stop codons and an amino acid composition based on the average ratios observed in natural, globular proteins, the search methods yielded similar nucleotide ratios, Semirandom DNA, synthesized with a designed, three-residue repeat pattern, can encode libraries of very high diversity and represents an important tool for the construction of random polypeptide libraries. PMID- 8401211 TI - Cloning, characterization, and expression of the beta subunit of pig heart succinyl-CoA synthetase. AB - The form of succinyl-CoA synthetase found in mammalian mitochondria is known to be an alpha beta dimer. Both GTP- and ATP-specific isozymes are present in various tissues. We have isolated essentially identical complementary DNA clones encoding the beta subunit of pig heart succinyl-CoA synthetase from both newborn and adult tissues. These cDNAs include a 1.4-kb sequence encoding the cytoplasmic precursor to the beta subunit comprised of 417 amino acid residues including a 22 residue mitochondrial targeting sequence. The cDNA encoding the 395-amino acid, 42,502-Da mature protein was confirmed to be the succinyl-CoA synthetase beta subunit by agreement with the N-terminal protein sequence and by high homology to prokaryotic forms of the beta subunit that were previously cloned (about 45% identical to beta from Escherichia coli). In contrast to a previous report (Nishimura, J.S., Ybarra, J., Mitchell, T., & Horowitz, P.M., 1988, Biochem. J. 250, 429-434), we found no tryptophan residue to be encoded in the sequence for the mature beta subunit, and this finding is corroborated by the fact that highly purified pig heart succinyl-CoA synthetase shows no tryptophan fluorescence or tryptophan content in amino acid compositional analysis. The cDNA clones encoding the mature pig heart beta subunit and its counterpart alpha subunit were coexpressed in a deletion mutant strain of E. coli. Recovery of succinyl-CoA synthetase activity demonstrated that this combination of subunits forms a productive enzymatic complex having GTP specificity. PMID- 8401212 TI - Conformational intermediates in the folding of a coiled-coil model peptide of the N-terminus of tropomyosin and alpha alpha-tropomyosin. AB - Circular dichroism was used to study the folding of alpha alpha-tropomyosin and AcTM43, a 43-residue peptide designed to serve as a model for the N-terminal domain of tropomyosin. The sequence of the peptide is AcMDAIKKKMQMLKLDVENLLDRLEQLEADLKALEDRYKQLEGGC. The peptide appeared to form a coiled coil at low temperatures (< 25 degrees C) in buffers with physiological ionic strength and pH. The folding and unfolding of the peptide, however, were noncooperative. When CD spectra were examined as a function of temperature, the apparent degree of folding differed when the ellipticity was followed at 222, 208, and 280 nm. Deconvolution of the spectra suggested that at least three component curves contributed to the CD in the far UV. One component curve was similar to the CD spectrum of the coiled-coil alpha-helix of native alpha alpha tropomyosin. The second curve resembled the spectrum of single-stranded short alpha-helical segments found in globular proteins. The third was similar to that of polypeptides in the random coil conformation. These results suggested that as the peptide folded, the alpha-helical content increased before most of the coiled coil was formed. When the CD spectrum of striated muscle alpha alpha-tropomyosin was examined as a function of temperature, the unfolding was also not totally cooperative. As the temperature was raised from 0 to 25 degrees C, there was a decrease in the coiled coil and an increase in the conventional alpha-helix type spectrum without formation of random coil. The major transition, occurring at 40 degrees C, was a cooperative transition characterized by the loss of all of the remaining coiled coil and a concomitant increase in random coil. PMID- 8401213 TI - Hydrophobic core repacking and aromatic-aromatic interaction in the thermostable mutant of T4 lysozyme Ser 117-->Phe. AB - The T4 lysozyme mutant Ser 117-->Phe was isolated fortuitously and found to be more thermostable than wild-type by 1.1-1.4 kcal/mol. In the wild-type structure, the side chain of Ser 117 is in a sterically restricted region near the protein surface and forms a short hydrogen bond with Asn 132. The crystal structure of the S117F mutant shows that the introduced Phe side chain rotates by about 150 degrees about the C alpha-C beta bond relative to wild type and is buried in the hydrophobic core of the protein. Burial of Phe 117 is accommodated by rearrangements of the surrounding side chains of Leu 121, Leu 133, and Phe 153 and by main-chain shifts, which result in a minimal increase in packing density. The benzyl rings of Phe 117 and Phe 153 form a near-optimal edge-face interaction in the mutant structure. This aromatic-aromatic interaction, as well as increased hydrophobic stabilization and elimination of a close contact in the wild-type protein, apparently compensate for the loss of a hydrogen bond and the possible cost of structural rearrangements in the mutant. The structure illustrates the ability of a protein to accommodate a surprisingly large structural change in a manner that actually increases thermal stability. The mutant has activity about 10% that of wild-type, supportive of the prior hypothesis (Grutter, M.G. & Matthews, B.W., 1982, J. Mol. Biol. 154, 525-535) that the peptidoglycan substrate of T4 lysozyme makes extended contacts with the C-terminal domain in the vicinity of Ser 117. PMID- 8401214 TI - Effect of pH and denaturants on the folding and stability of murine interleukin 6. AB - The conformation and stability of a recombinant mouse interleukin-6 (mIL-6) has been investigated by analytical ultracentrifugation, fluorescence spectroscopy, urea-gradient gel electrophoresis, and near- and far-ultraviolet circular dichroism. On decreasing the pH from 8.0 to 4.0, the tryptophan fluorescence of mIL-6 was quenched 40%, the midpoint of the transition occurring at pH 6.9. The change in fluorescence quantum yield was not due to unfolding of the molecule because the conformation of mIL-6, as judged by both urea-gradient gel electrophoresis and CD spectroscopy, was stable over the pH range 2.0-10.0. Sedimentation equilibrium experiments indicated that mIL-6 was monomeric, with a molecular mass of 22,500 Da over the pH range used in these physicochemical studies. Quenching of tryptophan fluorescence (20%) also occurred in the presence of 6 M guanidine hydrochloride upon going from pH 7.4 to 4.0 suggesting that an amino acid residue vicinal in the primary structure to one or both of the two tryptophan residues, Trp-36 and Trp-160, may be partially involved in the quenching of endogenous fluorescence. In this regard, similar results were obtained for a 17-residue synthetic peptide, peptide H1, which corresponds to an N-terminal region of mIL-6 (residues Val-27-Lys-43). The pH-dependent acid quenching of endogenous tryptophan fluorescence of peptide H1 was 30% in the random coil conformation and 60% in the presence of alpha-helix-promoting solvents. Replacement of His-33 with Ala-33 in peptide H1 alleviated a significant portion of the pH-dependent quenching of fluorescence suggesting that the interaction of the imidazole ring of His-33 with the indole ring of Trp-36 is a major determinant responsible for the quenching of the endogenous protein fluorescence of mIL-6. PMID- 8401215 TI - Defensins promote fusion and lysis of negatively charged membranes. AB - Defensins, a family of cationic peptides isolated from mammalian granulocytes and believed to permeabilize membranes, were tested for their ability to cause fusion and lysis of liposomes. Unlike alpha-helical peptides whose lytic effects have been extensively studied, the defensins consist primarily of beta-sheet. Defensins fuse and lyse negatively charged liposomes but display reduced activity with neutral liposomes. These and other experiments suggest that fusion and lysis is mediated primarily by electrostatic forces and to a lesser extent, by hydrophobic interactions. Circular dichroism and fluorescence spectroscopy of native defensins indicate that the amphiphilic beta-sheet structure is maintained throughout the fusion process. Taken together, these results support the idea that protein-mediated membrane fusion depends not only on hydrophobic and electrostatic forces but also on the spatial arrangement of the amino acid residues to form a three-dimensional amphiphilic structure, which promotes the efficient mixing of the lipids between membranes. A molecular model for membrane fusion by defensins is presented, which takes into account the contributions of electrostatic forces, hydrophobic interactions, and structural amphiphilicity. PMID- 8401216 TI - Independence of metal binding between tandem Cys2His2 zinc finger domains. AB - Most Cys2His2 zinc finger proteins contain tandem arrays of metal binding domains. The tandem nature of these arrays suggests that metal binding by these domains may not be independent but rather that metal binding may occur in a cooperative manner. This is especially true in light of the crystal structure of a three zinc finger array bound to DNA that revealed several types of interactions between domains. To address this question, peptides containing two tandem domains have been prepared. While metal binding studies do show that the two finger peptide has a metal ion affinity about threefold higher than that for a single domain peptide with the same sequence, additional studies reveal that this behavior is due to increased single site affinities in the context of the two domain peptide rather than to cooperativity. These studies indicate that domains of this type are independent of one another with regard to metal binding, at least in the absence of DNA. This observation has implications with regard to the question of whether the activities of proteins of this class might be modulated by available zinc concentrations. PMID- 8401217 TI - Functional consequences of mutations at the allosteric interface in hetero- and homo-hemoglobin tetramers. AB - A seminal difference exists between the two types of chains that constitute the tetrameric hemoglobin in vertebrates. While alpha chains associate weakly into dimers, beta chains self-associate into tightly assembled tetramers. While heterotetramers bind ligands cooperatively with moderate affinity, homotetramers bind ligands with high affinity and without cooperativity. These characteristics lead to the conclusion that the beta 4 tetramer is frozen in a quaternary R-state resembling that of liganded HbA. X-ray diffraction studies of the liganded beta 4 tetramers and molecular modeling calculations revealed several differences relative to the native heterotetramer at the "allosteric" interface (alpha 1 beta 2 in HbA) and possibly at the origin of a large instability of the hypothetical deoxy T-state of the beta 4 tetramer. We have studied natural and artificial Hb mutants at different sites in the beta chains responsible for the T-state conformation in deoxy HbA with the view of restoring a low ligand affinity with heme-heme interaction in homotetramers. Functional studies have been performed for oxygen equilibrium binding and kinetics after flash photolysis of CO for both hetero- and homotetramers. Our conclusion is that the "allosteric" interface is so precisely tailored for maintaining the assembly between alpha beta dimers that any change in the side chains of beta 40 (C6), beta 99 (G1), and beta 101 (G3) involved in the interface results in increased R-state behavior. In the homotetramer, the mutations at these sites lead to the destabilization of the beta 4 hemoglobin and the formation of lower affinity noncooperative monomers. PMID- 8401218 TI - Backbone assignments and secondary structure of the Escherichia coli enzyme-II mannitol A domain determined by heteronuclear three-dimensional NMR spectroscopy. AB - This report presents the backbone assignments and the secondary structure determination of the A domain of the Escherichia coli mannitol transport protein, enzyme-IImtl. The backbone resonances were partially assigned using three dimensional heteronuclear 1H NOE 1H-15N single-quantum coherence (15N NOESY-HSQC) spectroscopy and three-dimensional heteronuclear 1H total correlation 1H-15N single-quantum coherence (15N TOCSY-HSQC) spectroscopy on uniformly 15N enriched protein. Triple-resonance experiments on uniformly 15N/13C enriched protein were necessary to complete the backbone assignments, due to overlapping 1H and 15N frequencies. Data obtained from three-dimensional 1H-15N-13C alpha correlation experiments (HNCA and HN(CO)CA), a three-dimensional 1H-15N-13CO correlation experiment (HNCO), and a three-dimensional 1H alpha-13C alpha-13CO correlation experiment (COCAH) were combined using SNARF software, and yielded the assignments of virtually all observed backbone resonances. Determination of the secondary structure of IIAmtl is based upon NOE information from the 15N NOESY HSQC and the 1H alpha and 13C alpha secondary chemical shifts. The resulting secondary structure is considerably different from that reported for IIAglc of E. coli and Bacillus subtilis determined by NMR and X-ray. PMID- 8401219 TI - Crystallization of two integral membrane pigment-protein complexes from the purple-sulfur bacterium Chromatium purpuratum. PMID- 8401220 TI - An ATP- and hsc70-dependent oligomerization of nascent heat-shock factor (HSF) polypeptide suggests that HSF itself could be a "sensor" for the cellular stress response. PMID- 8401221 TI - Prediction of membrane-spanning beta-strands and its application to maltoporin. PMID- 8401222 TI - The joys and vicissitudes of protein science. PMID- 8401223 TI - Hematopoietic cytokines: similarities and differences in the structures, with implications for receptor binding. AB - Crystal and NMR structures of helical cytokines--interleukin-4 (IL-4), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-2 (IL 2)--have been compared. Root mean square deviations in the C alpha coordinates for the conserved regions of the helices were 1-2 A between different cytokines, about twice the differences observed for independently determined crystal and solution structures of IL-4. Considerable similarity in amino acid sequence in the areas expected to interact with the receptors was detected, and the available mutagenesis data for these cytokines were correlated with structure conservation. Models of cytokine-receptor interactions were postulated for IL-4 based on its structure as well as on the published structure of human growth hormone interacting with its receptors (de Vos, A.M., Ultsch, M., & Kossiakoff, A.A., 1992, Science 255, 306-312). Patches of positively charged residues on the surfaces of helices C and D of IL-4 may be responsible for the interactions with the negatively charged residues found in the complementary parts of the IL-4 receptors. PMID- 8401224 TI - Conformational instability of the N- and C-terminal lobes of porcine pepsin in neutral and alkaline solutions. AB - Pepsin contains, in a single chain, two conformationally homologous lobes that are thought to have been evolutionarily derived by gene duplication and fusion. We have demonstrated that the individual recombinant lobes are capable of independent folding and reconstitution into a two-chain pepsin or a two-chain pepsinogen (Lin, X., et al., 1992, J. Biol. Chem. 267, 17257-17263). Pepsin spontaneously inactivates in neutral or alkaline solutions. We have shown in this study that the enzymic activity of the alkaline-inactivated pepsin was regenerated by the addition of the recombinant N-terminal lobe but not by the C terminal lobe. These results indicate that alkaline inactivation of pepsin is due to a selective denaturation of its N-terminal lobe. A complex between recombinant N-terminal lobe of pepsinogen and alkaline-denatured pepsin has been isolated. This complex is structurally similar to a two-chain pepsinogen, but it contains an extension of a denatured pepsin N-terminal lobe. Acidification of the complex is accompanied by a cleavage in the pro region and proteolysis of the denatured N terminal lobe. The structural components that are responsible for the alkaline instability of the N-terminal lobe are likely to be carboxyl groups with abnormally high pKa values. The electrostatic potentials of 23 net carboxyl groups in the N-terminal domain (as compared to 19 in the C-terminal domain) of pepsin were calculated based on the energetics of interacting charges in the tertiary structure of the domain. The groups most probably causing the alkaline denaturation are Asp11, Asp159, Glu4, Glu13, and Asp118.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401225 TI - Site-specific mutations in the N-terminal region of human C5a that affect interactions of C5a with the neutrophil C5a receptor. AB - C5a is an inflammatory mediator that evokes a variety of immune effector functions including chemotaxis, cell activation, spasmogenesis, and immune modulation. It is well established that the effector site in C5a is located in the C-terminal region, although other regions in C5a also contribute to receptor interaction. We have examined the N-terminal region (NTR) of human C5a by replacing selected residues in the NTR with glycine via site-directed mutagenesis. Mutants of rC5a were expressed as fusion proteins, and rC5a was isolated after factor Xa cleavage. The potency of the mutants was evaluated by measuring both neutrophil chemotaxis and degranulation (beta-glucuronidase release). Mutants that contained the single residue substitutions Ile-6-->Gly or Tyr-13-->Gly were reduced in potency to 4-30% compared with wild-type rC5a. Other single-site glycine substitutions at positions Leu-2, Ala-10, Lys-4, Lys-5, Glu 7, Glu-8, and Lys-14 showed little effect on C5a potency. The double mutant, Ile 6-->Gly/Tyr-13-->Gly, was reduced in potency to < 0.2%, which correlated with a correspondingly low binding affinity for neutrophil C5a receptors. Circular dichroism studies revealed a 40% reduction in alpha-helical content for the double mutant, suggesting that the NTR contributes stabilizing interactions that maintain local secondary or tertiary structure of C5a important for receptor interaction. We conclude that the N-terminal region in C5a is involved in receptor binding either through direct interaction with the receptor or by stabilizing a binding site elsewhere in the intact C5a molecule. PMID- 8401226 TI - A study of the effects of altering the sites for N-glycosylation in alpha-1 proteinase inhibitor variants M and S. AB - alpha-1-Proteinase inhibitor (A1Pi) is a monomeric secreted protein glycosylated at asparagines 46, 83, and 247. For this study cDNAs for M (normal) and S (Glu264 ->Val) variants of A1Pi were altered by site-directed mutagenesis to produce the combinations of single, double, and triple mutants that can be generated by changing the codons normally specifying these Asn residues to encode Gln. The fates of the mutant proteins were followed in transiently transfected COS-1 cells. All variants with altered glycosylation sites are secreted at reduced rates, are partially degraded, accumulate intracellularly, and some form Nonidet P-40-insoluble aggregates. The carbohydrate attached at Asn83 seems to be of particular importance to the export of both A1PiM and A1PiS from the endoplasmic reticulum. All mutations affecting glycosylation of A1PiS notably reduce secretion, cause formation of insoluble aggregates, and influence degradation of the altered proteins. The variant of A1PiS missing all three glycosylation sites is poorly secreted, is incompletely degraded, and accumulates in unusual perinuclear vesicles. These studies show that N-linked oligosaccharides in A1Pi are vital to its efficient export from the endoplasmic reticulum and that the consequences of changing the normal pattern of glycosylation vary depending upon the sites altered and the variant of A1Pi bearing these alterations. PMID- 8401227 TI - Use of proline mutants to help solve the NMR solution structure of type III antifreeze protein. AB - To help understand the structure/function relationships in antifreeze proteins (AFP), and to define the motifs required for ice binding, a Type III AFP suitable for two-dimensional (2D) NMR studies was produced in Escherichia coli. A synthetic gene for one of the Type III AFP isoforms was assembled in a T7 polymerase-directed expression vector. The 67-amino acid-long gene product differed from the natural AFP by inclusion of an N-terminal methionine but was indistinguishable in activity. The NMR spectra of this AFP were complicated by cis-trans proline isomerization from the C-terminal sequence YPPA. Substitution of this sequence by YAA eliminated isomer signals without altering the activity or structure of the mutant AFP. This variant (rQAE m1.1) was selected for sequential assignment and the secondary structure determination using 2D 1H NMR spectroscopy. Nine beta-strands are paired to form two triple-stranded antiparallel sheets and one double-stranded antiparallel sheet. Two further proline replacements, P29A and P33A, were made to delineate the role of conserved prolines in Type III AFP. These mutants were valuable in clarifying ambiguous NMR spectral assignments amongst the remaining six prolines of rQAE m1.1. In contrast to the replacement of the C-terminal prolyl residues, the exchange of P29 and P33 caused some structural changes and significantly decreased protein solubility and antifreeze activity. PMID- 8401228 TI - The structure and function of omega loop A replacements in cytochrome c. AB - The structural and functional consequences of replacing omega-loop A (residues 18 32) in yeast iso-1-cytochrome c with the corresponding loop of Rhodospirillum rubrum cytochrome c2 have been examined. The three-dimensional structure of this loop replacement mutant RepA2 cytochrome c, and a second mutant RepA2(Val 20) cytochrome c in which residue 20 was back substituted to valine, were determined using X-ray diffraction techniques. A change in the molecular packing is evident in the RepA2 mutant protein, which has a phenylalanine at position 20, a residue considerably larger than the valine found in wild-type yeast iso-1-cytochrome c. The side chain of Phe 20 is redirected toward the molecular surface, altering the packing of this region of omega-loop A with the hydrophobic core of the protein. In the RepA2(Val 20) structure, omega-loop A contains a valine at position 20, which restores the original wild-type packing arrangement of the hydrophobic core. Also, as a result of omega-loop A replacement, residue 26 is changed from a histidine to asparagine, which results in displacements of the main-chain atoms near residue 44 to which residue 26 is hydrogen bonded. In vivo studies of the growth rate of the mutant strains on nonfermentable media indicate that the RepA2(Val 20) cytochrome c behaves much like the wild-type yeast iso-1 protein, whereas the stability and function of the RepA2 cytochrome c showed a temperature dependence. The midpoint reduction potential measured by cyclic voltammetry of the RepA2 mutant is 271 mV at 25 degrees C. This is 19 mV less than the wild-type and RepA2(Val 20) proteins (290 mV) and may result from disruption of the hydrophobic packing in the heme pocket and increased mobility of omega-loop A in RepA2 cytochrome c. The temperature dependence of the reduction potential is also greatly enhanced in the RepA2 protein. PMID- 8401229 TI - Recombinant human erythropoietin (rHuEPO): cross-linking with disuccinimidyl esters and identification of the interfacing domains in EPO. AB - Several amino groups of recombinant human erythropoietin are selectively cross linked by specific cross-linkers including disuccinimidyl suberate or dithiobis(succinimidyl propionate). Intramolecular cross-linkings are obtained without significant change of the protein conformation using appropriate concentrations (0.2 mM) of the cross-linkers, which possess an 11-12-A length of a spacer between two reacting groups. Intramolecularly cross-linked peptides obtained suggest that several amino groups in erythropoietin (EPO) are positioned at a distance of near 12 A in the solution state. These interfacing amino groups include Lys 20-Lys 154, Lys 45-Lys 140, Lys 52-Lys 154, Lys 52-Lys 140, and Ala 1 Lys 116. A comparison of the cross-linking results between nonglycosylated EPO and glycosylated EPO suggests that both proteins retain high similarity regarding protein conformation. These results fit a structural model similar to that of human growth hormone, in which four alpha-helical bundles and a long stretch of beta-sheet structure are involved in the active protein. PMID- 8401230 TI - Bacterial expression and characterization of the CREB bZip module: circular dichroism and 2D 1H-NMR studies. AB - In this paper we describe the expression and purification from bacteria of the recombinant basic leucine zipper (bZip) domain of the cAMP response element binding protein, CREB327. The bZip peptide, CREB259-327, purified to near homogeneity, maintains the sequence-specific CRE site recognition demonstrated by in vitro competition assays. Alkylation of the three cysteine residues of CREB259 327 was employed to prevent aggregation of the peptide due to cysteine oxidation. The Kd of the purified native and modified CREB259-327 for the CRE site was determined by gel retardation assays to be on the order of 10(-7) M. We employed CD spectroscopy to study the folding properties of the native and modified CREB259-327. The CD analyses of the native/modified CREB259-327 peptide demonstrated a 20% increase in the alpha-helical content upon binding to the cAMP response-element. Only a 5% increase in the alpha-helical content of CREB259-327 is observed upon binding to the AP-1 site. This observation contrasts with CREB from the GCN4 protein (Weiss, M.A., et al., 1990, Nature 347, 575-578). In addition, the two-dimensional (2D) 1H-NMR studies of the bZip CREB peptide further support the distinct features of the CREB protein, in comparison to GCN4. Analysis by CD and 2D NMR of the dimerization domain of CREB suggests that the distinct DNA binding characteristics of CREB reside in the basic portion of the bZip module. PMID- 8401231 TI - Role of the C-terminus in the activity, conformation, and stability of interleukin-6. AB - Two murine interleukin-6 (mIL-6) variants were constructed using the polymerase chain reaction (PCR), one lacking the last five residues (183-187) at the C terminus (pMC5) and another with the last five residues of mIL-6 substituted by the corresponding residues of human IL-6 (pMC5H). The growth stimulatory activity of pMC5 on the mouse hybridoma cell line 7TD1 was < 0.05% of mIL-6, whereas pMC5H and mIL-6 were equipotent. The loss of biological activity of pMC5 correlated with its negligible receptor binding affinity on 7TD1 cells, while the binding of pMC5H was comparable to that of mIL-6. Both pMC5 and pMC5H, like mIL-6, failed to interact with recombinant soluble human IL-6 receptor when assayed by surface plasmon resonance-based biosensor analysis. These studies suggest that the C terminal seven amino acids of human IL-6, alone, do not define species specificity for receptor binding. A variety of biophysical techniques, as well as the binding of a conformational-specific monoclonal antibody, indicated that the global fold of the mIL-6 variants was similar to that of mIL-6, although small changes in the NMR spectra, particularly for pMC5, were observed. Some of these changes involved residues widely separated in the primary structure. For instance, interactions involving Tyr-22 were influenced by the C-terminal amino acids suggesting that the N- and C-termini of mIL-6 are in close proximity. Equilibrium unfolding experiments indicated that pMC5 was 0.8 kcal/mol less stable than mIL-6, whereas pMC5H was 1.4 kcal/mol more stable. These studies emphasize the structural importance of the C-terminal amino acids of IL-6 and suggest that truncation or mutation of this region could lead to small but significant alterations in other regions of the molecule. PMID- 8401232 TI - Interactions and inhibition of blood coagulation factor Va involving residues 311 325 of activated protein C. AB - Activated protein C (APC) exerts its physiologic anticoagulant role by proteolytic inactivation of the blood coagulation cofactors Va and VIIIa. The synthetic peptide-(311-325) (KRNRTFVLNFIKIPV), derived from the heavy chain sequence of APC, potently inhibited APC anticoagulant activity in activated partial thromboplastin time (APTT) and Xa-1-stage coagulation assays in normal and in protein S-depleted plasma with 50% inhibition at 13 microM peptide. In a system using purified clotting factors, peptide-(311-325) inhibited APC-catalyzed inactivation of factor Va in the presence or absence of phospholipids with 50% inhibition at 6 microM peptide. However, peptide-(311-325) had no effect on APC amidolytic activity or on the reaction of APC with the serpin, recombinant [Arg358]alpha 1-antitrypsin. Peptide-(311-325) surprisingly inhibited factor Xa clotting activity in normal plasma, and in a purified system it inhibited prothrombinase activity in the presence but not in the absence of factor Va with 50% inhibition at 8 microM peptide. The peptide had no significant effect on factor Xa or thrombin amidolytic activity and no effect on the clotting of purified fibrinogen by thrombin, suggesting it does not directly inhibit these enzymes. Factor Va bound in a dose-dependent manner to immobilized peptide-(311 325). Peptide-(311-315) inhibited the binding of factor Va to immobilized APC or factor Xa.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401233 TI - Thermodynamics of apocytochrome b5 unfolding. AB - Apocytochrome b5 from rabbit liver was studied by scanning calorimetry, limited proteolysis, circular dichroism, second derivative spectroscopy, and size exclusion chromatography. The protein is able to undergo a reversible two-state thermal transition. However, transition temperature, denaturational enthalpy, and heat capacity change are reduced compared with the holoprotein. Apocytochrome b5 stability in terms of Gibbs energy change at protein unfolding (delta G) amounts to delta G = 7 +/- 1 kJ/mol at 25 degrees C (pH 7.4) compared with delta G = 25 kJ/mol for the holoprotein. Apocytochrome b5 is a compact, native-like protein. According to the spectral data, the cooperative structure is mainly based in the core region formed by residues 1-35 and 79-90. This finding is in full agreement with NMR data (Moore, C.D. & Lecomte, J.T.J., 1993, Biochemistry 32, 199-207). PMID- 8401234 TI - Modeling of protein loops by simulated annealing. AB - A method is presented to model loops of protein to be used in homology modeling of proteins. This method employs the ESAP program of Higo et al. (Higo, J., Collura, V., & Garnier, J., 1992, Biopolymers 32, 33-43) and is based on a fast Monte Carlo simulation and a simulated annealing algorithm. The method is tested on different loops or peptide segments from immunoglobulin, bovine pancreatic trypsin inhibitor, and bovine trypsin. The predicted structure is obtained from the ensemble average of the coordinates of the Monte Carlo simulation at 300 K, which exhibits the lowest internal energy. The starting conformation of the loop prior to modeling is chosen to be completely extended, and a closing harmonic potential is applied to N, CA, C, and O atoms of the terminal residues. A rigid geometry potential of Robson and Platt (1986, J. Mol. Biol. 188, 259-281) with a united atom representation is used. This we demonstrate to yield a loop structure with good hydrogen bonding and torsion angles in the allowed regions of the Ramachandran map. The average accuracy of the modeling evaluated on the eight modeled loops is 1 A root mean square deviation (rmsd) for the backbone atoms and 2.3 A rmsd for all heavy atoms. PMID- 8401235 TI - Verification of protein structures: patterns of nonbonded atomic interactions. AB - A novel method for differentiating between correctly and incorrectly determined regions of protein structures based on characteristic atomic interaction is described. Different types of atoms are distributed nonrandomly with respect to each other in proteins. Errors in model building lead to more randomized distributions of the different atom types, which can be distinguished from correct distributions by statistical methods. Atoms are classified in one of three categories: carbon (C), nitrogen (N), and oxygen (O). This leads to six different combinations of pairwise noncovalently bonded interactions (CC, CN, CO, NN, NO, and OO). A quadratic error function is used to characterize the set of pairwise interactions from nine-residue sliding windows in a database of 96 reliable protein structures. Regions of candidate protein structures that are mistraced or misregistered can then be identified by analysis of the pattern of nonbonded interactions from each window. PMID- 8401236 TI - Rapid protein separation and diffusion coefficient measurement by frit inlet flow field-flow fractionation. AB - In this study three flow field-flow fractionation (flow FFF) channels are utilized for the separation of proteins and for the simultaneous measurement of their translational diffusion coefficients, D. One channel has a traditional sample inlet, whereas the other two incorporate a frit inlet design that permits more convenient and rapid sample introduction. The dependence of retention time on D, which leads to differential elution and the opportunity to measure D for protein peaks purified by the flow FFF process, is described theoretically and examined experimentally. Factors affecting band broadening, resolution, and optimization are also examined. The separation of proteins is achieved in the time range 4-20 min. Partial resolution is achieved in multiple runs requiring 2 min each. Values of D calculated from retention times are reported for 15 proteins. These include two protein dimers (bovine serum albumin and gamma globulin) not ordinarily accessible to measurement. The D values from the three channels are compared with one another and with literature data. Reasonable consistency (within 3-4%) is found. High-speed repetitive runs can be used to acquire multiple values of D in time intervals as short as 1 min. PMID- 8401237 TI - Vibrio cholerae hlyB is a member of the chemotaxis receptor gene family. PMID- 8401238 TI - Growing up in the golden age of protein chemistry. PMID- 8401239 TI - Replicative studies. PMID- 8401240 TI - The impact of HIV-positive diagnosis on the individual, Part 1: Stigma, rejection, and loneliness. AB - The growing incidence of HIV infection and AIDS has generated panic, fear, anxiety, and negative attitudes among the general public. As an illness, HIV infection and AIDS have become the new sources of stigma. This article explores the stigma, rejection, and loneliness of 25 individuals with known HIV infection/AIDS, who were interviewed using a set of open-ended questions. The results indicated that the HIV-positive diagnosis had a profound impact on the individual's psychosocial aspects of life, particularly interactions with others. Most of those infected by the virus were lonely, had to cope with stigmatization, and suffered disruption of family and peer relationships. They were confronted with prolonged uncertainty about their lives and experienced intense, constant fear of disclosure of their illness and of being rejected by society, yet they were not always able to discuss these fears and anxieties openly with friends or family. PMID- 8401241 TI - Anxiety and the colposcopy experience. AB - The purpose of this study was to explore the anxiety of an abnormal Papanicolaou (Pap) result, the need for colposcopy, and the colposcopic examination experienced by first-time colposcopy clients. Spielberger's State-Trait Anxiety Inventory (STAI) and a Woman's Profile were administered to 149 women who had abnormal Pap smears necessitating colposcopy. The results confirm that this experience evokes anxiety. Younger women who knew less and had concurrent stressors in their lives were most anxious. Women identified ways that health professionals could be more helpful in alleviating their anxiety. Their suggestions included the provision of more information and an opportunity for personal contact with a health professional. PMID- 8401242 TI - Patterns of concerns and demands experienced by spouses following coronary artery bypass surgery. AB - The purpose of this study was twofold: first, to describe spouses' concerns and demands related to their partner's recovery from coronary artery bypass surgery (CABS) prior to discharge and at 1, 3, and 6 weeks following discharge; and second, to describe how spouses' concerns and demands change from the time of hospitalization through 6 weeks after discharge. The study used a descriptive panel design. Ninety females (82.6%) and 19 (17.4%) males constituted the sample. Respondents' mean age was 60.7 years. Manifest content analysis revealed that spouses' concerns and demands changed from week to week. The study findings support a convalescence trajectory and lend support for paying close attention to the timing of interventions after surgery and during early convalescence. PMID- 8401243 TI - Body and orofacial cathexis in edentulous complete-denture-wearing clients. AB - This description cross-sectional study examines cathexis, or satisfaction, with the body and the orofacial region in relationship to eight major demographic and health status variables in a nonprobability sample of 34 male and 47 female edentulous subjects aged 36 to 77 years. Kiyak's 27-item Body Cathexis Scale (BCS), including an innovative five-item Orofacial Cathexis Scale (OFCS), was shown to be reliable for use in elderly clients at least to age 77 (Cronbach's alpha = .95). More research is needed to validate the OFCS. Overall scores were in the neutral range, with teeth being the most negatively cathected body part. Longer periods of edentulism were associated with more dissatisfaction with the teeth. Body and orofacial cathexis were highly positively correlated. Chronic illness, education level, race, and religion were not significantly related to cathexis. Edentulism, especially recent edentulism in the elderly, female gender, and divorced marital status could be important indicators for body image disturbance. PMID- 8401244 TI - Experiences of primiparous breast-feeding mothers in the first days following birth. AB - This study describes the breast-feeding patterns of 59 neonates and the experiences of their primiparous mothers during the early postpartum period. The results showed that most mothers and babies in this group had significant difficulties during the first 2 days after birth and that 33% of the mother-baby pairs were still having problems latching or sucking on discharge from hospital (fourth to fifth postpartum days). Of the mothers who were still having problems on leaving the hospital, 84% had given up breast-feeding by 8 weeks, most of them by 2 weeks following hospital discharge. PMID- 8401245 TI - Behavioral analysis and behavioral strategies to improve self-management of type II diabetes. AB - The implications of behavioral analysis for practice and research have significant potential for nursing. This present study was conducted to determine the effectiveness of nurses and patients actively participating in behavioral analysis and the implementation of behavioral strategies in order to improve the patients' self-management of their Type II diabetes. Patients (N = 156) were randomly assigned to one of four groups. The attention control group (n = 41) received routine care. The compliance group (n = 32) agreed to practice compliance behaviors related to the prescribed medical regimen. The behavioral strategies group (n = 42) participated in behavioral analysis and agreed to practice behavioral strategies. The behavioral strategies with instruction group (n = 41) participated in behavioral analysis, agreed to practice behavioral strategies, and received classes and programmed instruction about behavioral analysis and behavioral strategies. There were no outcome differences between groups relative to glycosylated hemoglobin (GHb) and weight loss. There were differences in the outcome measures in subgroups by age, gender, and employment, which have practice and research implications for the individualization of interventions using behavioral strategies. PMID- 8401246 TI - Toward a definition of a successful dieter. AB - The purpose of the research was to examine the characteristics of successful weight losers and to compare the findings with a control group of unsuccessful weight losers. The project was designed around a definition of successful dieters, based on the Metropolitan Life Insurance weight tables. The definition, in fact, was not particularly useful. This article describes the difficulties associated with using the original definition as the inclusion criteria for sample selection, the reanalysis of data using the Body Mass Index (BMI), and finally, the creation of a new definition developed from the construction of weight history based on changes in the BMI as an adult. This article presents the final outcome of what a definition of a successful dieter should consist in future research. PMID- 8401247 TI - Adverse events in the hospitalized elderly. AB - This descriptive study examined the incidence, onset, and correlates of adverse events (acute confusion, adverse drug reactions, nutritional problems, pressure ulcers, falls, and transient urinary incontinence) experienced by elderly persons hospitalized for acute illness. The researchers studied 64 patients (mean age 74.4 years) for the duration of their hospitalization. Over half (54%) experienced some degree of acute confusion during their hospitalization. There was a 24% incidence of pressure ulcers and a 2% incidence of adverse reactions to medications. No instances of falls or transient urinary incontinence were observed or documented in this sample. Nutritional problems were identified in one third of the sample. Predictive profiles of patients at risk for adverse events associated with hospitalization were developed and provide directions for patient care and further research. PMID- 8401248 TI - An appraisal of strategies to reduce the incidence of breast cancer. AB - The current focus of breast cancer research is to develop a novel strategy to prevent the disease. In this review a potential model of breast cancer development is proposed based upon the results of laboratory models of the induction of mammary carcinogenesis. It is clear that susceptibility to initiation occurs in young female animals, and a preventive strategy is more effective the sooner it is started after initiation occurs. In humans we do not know the timing or the nature of the carcinogenic insult, but epidemiologic studies suggest that the process is long and initiation is most likely to occur in young adults. Hormones are the key to promotion of the carcinogenic process and it would appear that strategically the earlier an intervention is applied after initiation the better will be the general effect on the population. Hormonal contraception could prevent breast cancer if the appropriate formulation was chosen and used by all young women. This inhibitory strategy might protect women without the need to preselect based on risk factors. Breast cancer prevention would be a side effect of the contraceptive method. Alternatively, tamoxifen, an antiestrogen, is known to prevent mammary carcinogenesis in animals and prevent the appearance of second primary breast cancers in women. This well tested therapeutic agent is currently being evaluated in clinical trials of selected high-risk women aged 35 and above. Finally, retinoids have shown promise as agents in the laboratory to prevent cell replication and inhibit mammary tumorigenesis. A trial of retinoids to prevent second primary tumors in node negative breast cancer patients is currently underway in Italy. The review discusses the relative merits and concerns about these prevention strategies and proposes additional studies to be undertaken. PMID- 8401249 TI - Interleukin 2 treatment in acute myelogenous leukemia. AB - Significant clinical responses obtained with interleukin 2 (IL-2) in solid tumors such as renal cell cancer and malignant melanoma prompted the use of this immunomodulatory drug to verify its activity in hematological malignancies. Several preclinical experiments showed an activity of IL-2 against leukemic cell lines in cultures, particularly in acute myeloid leukemia (AML), while only episodically a proliferative stimulus of IL-2 on the growth of leukemic blasts has been observed. Based on these preclinical studies, in the past five years several phase I-II clinical trials have verified IL-2 activity in AML in advanced phase, both in patients with active disease and in patients in further complete remission (CR). Data obtained are difficult to evaluate due to the low number and the heterogeneity of patients treated, but encouraging results have been reported in patients with "limited" disease (bone marrow blastosis < 30%), showing an antileukemic activity of IL-2 alone. Different international phase III trials are ongoing in AML patients in I CR after autologous bone marrow transplantation (Roussel-Uclaf, Romainville, France) and in II CR after conventional chemotherapy (Roche SpA, Milan, Italy) to verify the efficacy of IL-2 in reducing the risk of relapse and prolonging disease-free survival. PMID- 8401250 TI - Roferon-A (interferon alpha 2a) combined with Tigason (etretinate) for treatment of cutaneous T cell lymphomas. AB - Many treatments are used for epidermotropic cutaneous T cell lymphomas (CTCL) such as mycosis fungoides (MF) and Sezary syndrome (SS). All pretend to be effective, but none is really curative. As single drug therapy provides a response rate of about 55% with interferon alpha and about 45% with etretinate, we studied the effectiveness of combining these two drugs for immunomodulatory therapy in epidermotropic CTCL. A review of four reports, including a multicenter study performed in 45 patients, indicates a response rate of 56%, with better results for MF than SS. Side effects are generally moderate when low doses are used. The mechanism of action of this combined therapy on cutaneous lesions remains unclear. In vitro, a synergistic effect of retinoids on interferon alpha antiviral activity has been demonstrated, and in vivo an immunohistochemical study showed that the combined therapy modulates antigens expressed by keratinocytes and increases cytotoxic cells in dermis without modifying the number of Langerhans cells in epidermis. PMID- 8401251 TI - Neuroblastoma: the result of multistep transformation? AB - Genetic and molecular abnormalities, in association with malignant phenotypes, have been previously demonstrated in a variety of human tumors. Although the multistep theory fits well for some cancers such as retinoblastoma and colon carcinoma, for many others it still remains to be proven. Neuroblastoma, a tumor found in pediatric patients, seems to fall into the multistep model. Nonrandom chromosome abnormalities have been found with 1p deletion, loss of heterozygosity for short arm of chromosome 1 and for chromosome 11q and 14q. Amplification of N myc oncogene and an increased level of Ras protein have also been demonstrated. Therefore, even if it is not possible to show that these mutations happen as discrete events in their order of appearance, the multistep model seems involved in neuroblastoma development. Neuroblastoma has a peculiar aspect, however, that makes this tumor a natural model of defect of cell differentiation. In fact, there is a particular subset of metastatic tumors that show spontaneous regression. In vitro, neuroblastoma cell lines can be induced to differentiate along the neural pathway using retinoic acid. Other natural and chemical substances are also able to induce cell differentiation. During retinoic acid treatment, N-myc oncogene expression decreases and other genes are deregulated. p53 and MDR1 gene expression increases. These two different aspects, failure of cell differentiation pathway and genetic mutations, make the neuroblastoma one of the most difficult problems of modern molecular biology. PMID- 8401252 TI - Impaired response of polycythemic mice to erythropoietin induced by protein starvation imposed after hormone administration. AB - The present study was performed to determine the stage of the erythropoietic pathway which is affected by starvation or protein deprivation and whose manifestation is a depressed response to exogenous erythropoietin (EPO). The response to recombinant human EPO was measured in post-hypoxic polycythemic mice by determination of 59Fe uptake into red cells, spleen and femur and/or erythroid colony forming units (CFU-E) and erythroid precursor cell concentrations in femoral marrow. Experimental mice were either starved or fed one of seven different diets whose protein (casein) content ranged from 0 to 20%. All diets were isocaloric. The response of mice maintained on the standard diet (Purina Lab chow) was taken as the normal one. Starvation during the 48-hour period immediately before EPO injection had no effect on the response to the hormone. Starvation, and protein deprivation to a lesser extent, during the 48-hour period following EPO, on the other hand, significantly reduced the response. There was a progressive increase in the response as the casein content of the diet was increased. A normal response was observed when dietary casein concentration was 10%. These findings indicate that nutritional deprivation or dietary protein alterations during the period immediately following EPO injection in polycythemic mice can have detrimental effects on the erythroid response in a model in which nutritional deprivation was relatively short and acute. They also indicate that the subnormal response is not due to a decreased size of the erythroid progenitor pool available for differentiation but to deficient rates of differentiation of erythropoietic units. PMID- 8401253 TI - Hemoregulatory peptide (HP5b) dimer effects on normal and malignant cells in culture. AB - The dimer of the hemoregulatory peptide HP5b has been investigated for biological effects on various cell types in culture including mouse granulocyte-macrophage colony forming units (CFU-GM) from agar and murine long-term bone marrow culture (LTBMC). While CFU-GM were significantly stimulated in both systems, mitogen activation of mouse T, B and natural killer (NK) cells was not affected. Peptide treated mouse 3T3 fibroblasts reached a higher saturation density than controls; otherwise no effect was seen. A series of malignant cell lines was also tested. On a human glioblastoma cell line (GaMg) and rat glioma cell line (BT5C) a slight but significant stimulatory effect was found, while human mammary carcinoma cells (MCF7) were not affected. On SC1 mouse lymphoma cells a slight stimulation of cell growth was seen during the first part of exponential growth. Since HP5b acts as a stimulator for stromal cell secretion of other growth factors, supernatants from a human bone marrow stromal cell line stimulated with HP5b were tested on various cell lines. The effects of the supernatants on cell growth of the tested cell lines were not affected by HP5b treatment. Taken together with available in vivo data, the results indicate that the hemoregulatory peptide is a selective stimulator of myelopoiesis. PMID- 8401254 TI - Glucocorticoid-induced growth inhibition and differentiation of a human megakaryoblastic leukemia cell line: involvement of glucocorticoid receptor. AB - A human megakaryoblastic leukemia cell line, HIMeg, was established recently. Previous studies have shown that retinoic acid (RA) and 1,25-dihydroxyvitamin D3 [1,25(OH)2 D3] have potent effects on the proliferation and differentiation of HIMeg cells. Recently, the effect of dexamethasone (Dex), a synthetic glucocorticoid, on HIMeg cell growth and differentiation was examined in comparison with RA and sex steroid hormones. It was observed that in a methylcellulose culture system, Dex suppressed the clonal proliferation of HIMeg cells in a dose-dependent manner. The inhibitory effects of Dex could be reversed by RU486, a potent glucocorticoid antagonist. In contrast, sex steroid hormones had little effect on the clonal proliferation of HIMeg cells. In a liquid culture system, only 2% of HIMeg cells expressed glycoprotein IIb/IIIa (GPIIb/IIIa) antigen without hormone treatment, whereas 45% and 30% of the cells expressed GPIIb/IIIa following the addition of RA and Dex, respectively. To examine the molecular basis of the hormone-induced cell differentiation, glucocorticoid receptor (GR) expression was studied by Scatchard analysis. It was shown that there existed a saturable, high-affinity GR in HIMeg cells and the GR number was altered after Dex or RA treatment of the cells, suggesting that the cellular effects of glucocorticoids on HIMeg cells were mediated by GR. PMID- 8401255 TI - On control of the hematopoietic cell proliferation. AB - Hematopoietic cells, detected as spleen colony forming units (CFU-S), vary in their proliferation rate, and this can be investigated in vivo by determining the fraction of CFU-S synthesizing DNA. A large number of measurements of CFU-S number and the fraction synthesizing DNA has been obtained under standardized conditions in normal mice and after a single dose of arabinosyl cytosine (ara-C). These data are analyzed with respect to the correlation between the number of CFU S and their DNA-synthesizing fraction since, in the past, it has been maintained that the CFU-S proliferation rate responds sensitively to changes in CFU-S numbers. The present analysis does not support this traditional view--it instead demonstrates that natural variations in CFU-S numbers occurring in the same range as those following ara-C do not trigger CFU-S proliferation. On the other hand, administration of ara-C triggers most of the surviving CFU-S into the cell cycle, while decreasing CFU-S numbers by only about 40%. Data are also presented showing changes of CFU-S numbers and their DNA-synthesizing fraction in mice given a single dose of cyclophosphamide (CY). CY reduces CFU-S numbers, but the fraction synthesizing DNA can be either increased or decreased depending on time after treatment. Both these experimental situations argue against a close relationship between CFU-S numbers and their proliferation rate and suggest a different or a more complex regulation. PMID- 8401256 TI - 5-fluorouracil permits access to a primitive subpopulation of peripheral blood stem cells. AB - Peripheral blood stem cells (PBSC) contain a mixture of mature and immature hematopoietic progenitors. Resistance to 5-Fluorouracil (5-FU) has been used to identify and characterize primitive quiescent stem cells among bone marrow (BM) cells. To see if the same technique could be used to isolate a similar population of cells among PBSC, low-density peripheral blood mononuclear cells (PBMNC) were collected by cytapheresis in the regenerative phase after high-dose chemotherapy from patients with hematological malignancies. These PBMNC were incubated with increasing concentrations of 5-FU for 24 h. The viable 5-FU resistant cells were then cultured in semi-solid media in the presence of either single cytokines: TCM 5637, Granulocyte Macrophage Colony Stimulating Factor (GM-CSF), or a combination of cytokines: interleukin 1 (IL-1) IL-1 + IL-3 + 5637, IL-1 + IL-3 + Stem Cell Factor (SCF). Low concentrations (5-10 micrograms/ml 5-FU) eliminated mature day 7 Colony Forming Units-Granulocyte Macrophage (CFU-GM) and spared day 7 clusters while enriching for day 14 CFU-GM, irrespective of the growth factors used. Higher concentrations of 5-FU (15, 20, 25 micrograms/ml) selected for later forming clonogenic elements. A combination of synergistic growth factors was required for the development of morphologically identifiable clonogenic elements resistant to 25 micrograms/ml 5-FU at day 21 of culture. Further experimentation demonstrated that SCF could effectively replace TCM 5637 in the cytokine combination for the detection of primitive late forming clonogenic elements. The presence of SCF potentiated colony formation by 5-FU resistant PBMNC. It was confirmed that GM-CSF alone was unable to support colony formation by PBMNC resistant to 25 micrograms/ml. These observations demonstrate that PBSC contain a heterogenous mixture of hematopoietic progenitors and that incubation with 25 micrograms/ml 5-FU permits access to a quiescent primitive stem cell population that requires a combination of synergistic growth factors for the development of morphologically identifiable clonogenic elements at day 21. Taken together, these results suggest that PBSC have similar characteristics to BM derived stem cells. PMID- 8401257 TI - Production of interleukin 6, leukemia inhibitory factor and granulocyte macrophage colony stimulating factor by peripheral blood mononuclear cells in Fanconi's anemia. AB - The aim of this study was to test the in vitro cytokine production by peripheral blood mononuclear cells (PBMCs) in patients with Fanconi's anemia (FA). Phytohemagglutinin (PHA)-stimulated PBMCs from 21 patients with FA were studied for their ability to produce interleukin 6 (IL-6), leukemia inhibitory factor (LIF) and granulocyte-macrophage colony stimulating factor (GM-CSF). Enzymatic immunoassay (EIA) was used for both IL-6 and LIF, while GM-CSF was evaluated in a highly sensitive biological assay provided by GM-CSF-dependent M-07e cells. A significant decrease of IL-6 was detected in 9 out of 11 FA patients compared with five normal donors, while similar amounts of LIF were produced from 21 FA patients and 21 healthy subjects. A drastic increase of active GM-CSF was documented in PHA-stimulated PBMC-conditioned medium in all 18 FA patients tested. Since IL-6 and GM-CSF play an important role in maintaining basal hemopoiesis, our results suggest that an abnormal cytokine network may be involved in the pathogenesis of FA pancytopenia. PMID- 8401258 TI - The biology of interleukin 11. AB - Interleukin 11 (IL-11) is a multifunctional cytokine which may play a role in regulating the growth and development of cells in both the hematopoietic and lymphoid systems. IL-11 activity was originally detected in the conditioned medium of a primate bone marrow stromal cell line, and the human cDNA was cloned from a human fetal lung fibroblast cell line. The purified protein shows multifunctional activity, influencing lymphohematopoietic stem cell proliferation and differentiation, megakaryocyte progenitor cell proliferation and differentiation, erythroid progenitor cell proliferation, B lymphocyte maturation, activation of hepatocyte acute phase protein synthesis, and adipogenesis. At the molecular level, IL-11 is unique, containing no asparagine linked glycosylation sites and no cysteine residues. The IL-11 receptor belongs to a family of cytokine receptors which includes the receptors for IL-6, leukemia inhibitory factor (LIF), oncostatin M (OSM), and ciliary neurotrophic factor (CNTF), which are all capable of interacting with the signal transducing receptor gp130 after ligand binding. IL-11 has demonstrated activity in preclinical models for the treatment of thrombocytopenia and, in some cases, neutropenia; studies are underway to confirm its usefulness in the clinic for treatment of myelosuppression associated with cancer chemotherapy and bone marrow transplantation. PMID- 8401259 TI - The hepatocyte growth factor and its receptor. AB - Hepatocyte growth factor/scatter factor (HGF/SF) is secreted by cells of mesodermal origin and shows powerful mitogenic, motogenic and morphogenic activities on epithelial and endothelial cells. It is a heparin-binding polypeptide with an alpha/beta heterodimeric structure, showing structural homologies with enzymes of the blood clotting cascade. HGF binds with high affinity to the receptor encoded by the MET protooncogene (p190MET). The MET receptor is a heterodimer of two disulfide-linked subunits (alpha and beta); the alpha subunit is extracellular, while the beta is transmembrane and endowed with tyrosine kinase activity. The HGF-triggered signalling is mediated by different cytoplasmic effectors, including phosphatidylinositol 3-kinase, phospholipase C gamma, and Src-related tyrosine kinases. p190MET is expressed in several normal epithelial tissues (e.g., liver, gastrointestinal tract, kidney) and is often overexpressed in neoplastic cells. p190MET expression has been reported also in central nervous system microglia, a monocyte-derived cell population. We recently found that p190MET is expressed in selected peripheral blood cell populations, such as macrophages. The amount of both mRNA and protein is barely detectable, while it is dramatically increased upon activation. These findings suggest that HGF may play a role in hemopoietic cell signaling, during activation and differentiation of blood cell lineages. PMID- 8401260 TI - Granulocyte-macrophage colony stimulating factor and interleukin 3: target cells and kinetics of response in vivo. AB - Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 3 (IL 3) target cells have been studied in vivo in subjects with normal hemopoiesis. GM CSF administration elicits a rapid and sustained neutrophilia, monocytosis and eosinophilia due to a direct proliferative stimulus on all progenitors and precursors of the granulomonopoietic lineage. GM-CSF is also a powerful stimulator of erythroid burst forming unit (BFU-E) and megakaryocyte colony forming unit (CFU-MK) proliferation. Its action, however, does not extend to more mature erythroid and megakaryocyte cells suggesting the need for combined treatment with lineage-specific growth factors such as erythropoietin (Epo) or IL 6 to obtain a complete myeloid stimulation. When GM-CSF is discontinued, its action rapidly vanishes, and a rapid decline in the proliferative rate of target cells to values below the initial ones occurs. The potential clinical usefulness of this phenomenon in regard to cancer chemotherapy is discussed. IL-3 treatment induces only a rapid and marked eosinophilia. Chronic IL-3 administration, however, increases the proliferation of all myelopoietic progenitors and primes CFU-GM to become more sensitive in vitro to the action of granulocyte CSF (G CSF), GM-CSF and IL-5. Whereas an increased IL-5 sensitivity seems devoid of therapeutic potential, the priming of G-CSF and GM-CSF action suggests rational scheduling for a combined treatment of IL-3 with other hemopoietic growth factors. PMID- 8401261 TI - Effect of recombinant IL-3 on lymphocyte populations in irradiated mice. AB - Administration of murine recombinant interleukin 3 (IL-3) to sublethally irradiated mice induced in the thymus recovery of the cell count and mitotic responsiveness to Con A, as well as a decrease in CD4-CD8- cells concomitant with an increase in single positive cells. Also in the spleen, the cell count and mitotic responsiveness to Con A and lipopolysaccharide (LPS) were recovered by IL 3 treatment. These findings show that IL-3 induces differentiation and growth of thymocytes and recovery of T and B cell functions in sublethally irradiated mice. PMID- 8401262 TI - Abnormal thermal hyperaemia in the skin in paraplegia. AB - Surface insulation, together with laser Doppler flowmetry, was used to assess the skin microcirculation of paraplegic patients. Two control groups of five male and five female subjects were used to establish the response of normals with which to compare the results obtained from six paraplegic subjects. No significant sex related difference was revealed from this study. It was found that in normal subjects, surface insulation resulted in a significant increase in both skin temperature and skin blood flow. In paraplegic patients, the temperature increase was significantly less than in the normal subjects and there was no significant thermally induced hyperaemia after surface insulation. PMID- 8401263 TI - A preliminary assessment of laser Doppler perfusion imaging in human skin using the tuberculin reaction as a model. AB - Laser Doppler imaging is a new development in the field of laser Doppler flowmetry. We were recently loaned the Lisca laser Doppler perfusion imager (LDI) manufactured by Lisca Development, Linkoping, Sweden for assessment against other methods for assessment of skin perfusion. In order to evaluate the device, it was applied to imaging flux changes induced in human skin during the tuberculin reaction. Flux values were compared directly with those from conventional lightguide laser Doppler flowmetry, and parameters measured using two other methods for assessment of tissue perfusion, dynamic thermographic imaging and tissue spectrophotometry. The results showed very good correlations between the various methods. In addition, the LDI showed that very large differences in flux values (up to 5 V) could be found within distances of only 2 mm during the tuberculin reaction and that focal centres of low flux values surrounded by higher values ('craters') could be found not only at the centre, but elsewhere in the lesion. The LDI enables rapid non-invasive detailed analysis of blood flow patterns in skin and correlates well with other methods for measuring skin perfusion. Its use to examine heterogeneity of microvascular blood flow patterns may lead to further understanding of the local mechanisms for regulation of oxygen supply to tissue. PMID- 8401264 TI - Time-frequency analysis of infant cry: measures that identify individuals. AB - As part of a larger study designed to test the predictive power of recordings of infant cry for neurological development at a year of age, we have developed a summary measure of a cry which shows a very high consistency within an individual. This measure may be useful in making assessments of anatomical structure in the infant vocal tract. PMID- 8401265 TI - Energy distribution in the spectrograms of the cries of normal and birth asphyxiated infants. AB - This paper describes the distribution of energy and energy variance with frequency in the cries of normal and birth asphyxiated infants recorded within eight days of delivery. Single-variable statistical analysis suggested that asphyxiated infants have their cries shifted up in frequency compared to control infants, up to a frequency of 10 kHz. PMID- 8401266 TI - Measurement of trabecular bone mineral density in the distal radius by two gamma ray computed tomography scanners. AB - The precision and measurement agreement of two gamma-ray computed tomography forearm bone scanners have been evaluated. The inherent precision in vitro for the Stratec SCT 900 scanner was 0.28% compared with 0.04% for the OSCAR scanner. The measurements were linear over the range of densities found in humans. Increasing thickness of the cortical shell dramatically increased trabecular bone attenuation coefficient (TBAC) for the SCT 900, but only slightly affected results with the OSCAR. Short-term precision of the SCT 900 in 22 women (1.26%) was not as good as that previously reported for the OSCAR (0.50%). The measured TBAC for the same patients was significantly greater with the SCT 900. Corrections for the error introduced by the cortical shell of the radius with the SCT were calculated. Using this correction similar TBAC values could be obtained with both scanners, but the variance of the differences was too great for the results to be used interchangeably. Published bone density data for normal subjects indicate that Scottish women lose more trabecular bone in the two decades after 50 years of age than those in the Federal Republic of Germany. PMID- 8401267 TI - Elasticity and geometry measurements of vascular specimens using a high resolution laboratory CT scanner. AB - Vascular diseases are frequently associated with changes in the mechanical properties of the arterial wall. Existing techniques for studying arterial geometry and mechanical properties in vitro are often destructive, since they involve sectioning of the specimen into strips, or provide average measurements of the mechanical properties over the volume of intact specimens. We developed a high-resolution computed tomography (CT) scanner for in vitro studies of arterial geometry and static elastic properties. The x-ray image intensifier based system can acquire single transverse images, or a volume image, with 2 mm-1 resolution. Images were obtained through an intact abdominal aortic aneurysm at five pressures. The incremental circumferential Young's modulus E(inc) was calculated from the internal and external circumferences, and at physiological pressures E(inc) of the aneurysm was found to be 275 times greater than that of the normal aorta proximal to it. A volume image of the specimen provided landmarks that allowed histological sections to be obtained at locations coincident with those where the elasticity was measured. The histological analysis revealed a sixfold decrease in elastin content in the aneurysm, compared to the normal aorta. We have demonstrated that the static mechanical properties and geometry of vascular specimens can be quantified in vitro with the new high-resolution CT scanner and can be compared subsequently with histological analysis to provide further insight into the understanding of atherogenesis. PMID- 8401268 TI - Blood vessel diameter measurement by ultrasound. AB - Blood vessel diameter measurement is crucial to volume flow measurement in medical ultrasound. The accuracy of the measurement is examined in this study. Experiments were made on three kinds of tube using four transducers on three commercial duplex ultrasound scanners. The accuracy of the inner diameter measurement is affected by ultrasonic pulse length. By correcting this effect, it has been shown that the accuracy of the measurement can be increased significantly over a range of conditions. PMID- 8401269 TI - Electrical impedance and haematocrit of human blood with various anticoagulants. AB - The relationship between the electrical impedance parameters and the haematocrit of normal human blood has been determined at 37 degrees C in the haematocrit range of 20-60% for four types of anticoagulant: ACD, EDTA, sodium heparin and buffered sodium citrate. For plasma resistance Rp and interior resistance R(i), the measured data could be fitted by both the exponential expression and the Maxwell-Fricke expressions, Rp = a/(1-h) + b and R(i) = a/h + b, where a and b are constants and h is the haematocrit expressed as a decimal. The membrane capacitance Cm was found to be directly proportional to the haematocrit and could be normalized to 100% haematocrit by the Fricke formula, C100 = C(m)/(1-rho p/Rp), where rho p is the resistivity of plasma. Observed differences for Rp and C(m) among samples with various anticoagulants indicate that the type of anticoagulant influences the electrical properties of the red cells and the plasma. PMID- 8401270 TI - Fractal analysis of foetal heart rate variability. AB - Current methods for analysing foetal heart rate (FHR) patterns have yet to meet their full potential in the recognition of hypoxia in the foetus. Following the recent suggestion that fractal analysis can be applied to FHR recordings, the current paper describes a method for distinguishing two simultaneous fractal dimensions in FHR variation. An irregular line was plotted from 2500 consecutive foetal heart beat to beat intervals derived from an ultrasound source. A window of 500 intervals was moved along the line in steps of 20 intervals. At each step the Richardson technique was used to make estimates of the length of the line within the window using 40 different ruler lengths. When the estimates were plotted against the ruler lengths on log-log axes the resulting curve exhibited two distinct linear regions, each demonstrating an inverse power relationship. From the two slopes the fractal dimensions were derived for unspecified low- and high-frequency FHR variation in the current window. The values of both fractal dimensions were plotted simultaneously with the irregular FHR line and were found to accord with perceived changes in FHR variation. The method described is simply a measure of the irregularity in a series of foetal heart beat to beat intervals: the existence of fractal properties in the irregular line does not of itself imply underlying deterministic dynamics (e.g. chaos). This new method of observing FHR variability requires no preprocessing of the measured data, which are all taken into account. Not only does it represent a method for studying normal foetal behaviour but also has potential as a sensitive indicator of impending foetal compromise. PMID- 8401271 TI - Apparatus to measure the step and frequency responses of gas analysis instruments. AB - The dynamic characteristics of gas analysers are often assessed by measuring the step response. It is difficult to generate a verifiable instantaneous step change in gas composition. We constructed a 0.06 ml measurement chamber connected via high-speed valves (0.5 ms response time) to two 31 reservoirs pressurized to 50 kPa with gases containing different concentrations of CO2. An electronic system opens the valves alternately depending on the polarity of a control voltage Vc. Two walls of the chamber contain narrow-band infra-red filters centered at 4.24 microns (50% transmission points at 4.16 and 4.32 microns) where CO2 absorption is high. A photoconductive infra-red sensor and an infra-red source are positioned on either side of the chamber. The output of the sensor is amplified by an instrumentation amplifier. Signal averaging of the sensor output in either the time or frequency domain was used to overcome the noise of the infra-red sensor. Step changes in Vc yielded exponentially changing outputs with a time constant of 1.1 ms. A quadrupole mass spectrometer's response to step changes in CO2 concentration generated in the measurement chamber fitted single exponential curves well with a maximum time constant of 37.7 ms and transport delay of 194 ms. The frequency response of the infra-red system, from Vc to the sensor output, fell by 0.7 dB with a phase lag of 30 degrees between 1 and 50 Hz. Using the infra-red system to measure the true input to the mass spectrometer, the frequency response of the mass spectrometer was found to fall by 35 dB with a phase lag of over 3000 degrees between 0.2 and 50 Hz. A first-order model with delay fitted to the step response predicted the mass spectrometer frequency response well below 10 Hz but overestimated the response above 10 Hz. A third order model with delay fitted to the frequency response predicted the step response very well. Our results suggest that low-order models cannot predict the high-frequency performance of a mass spectrometer. PMID- 8401272 TI - The development of a technique to measure bone aluminium content using neutron activation analysis. AB - The accumulation and toxicity of aluminium in patients with chronic renal failure is a well recognized hazard, and there is a need for a non-invasive technique to assess Al tissue load in these patients. The technique of in vivo neutron activation analysis, using a thermal neutron beam from a reactor, has been employed by previous workers, who measured Al in the hand with a detection limit of 0.4 mg for a dose equivalent of 20 mSv. However, the application of this technique is restricted by the very limited availability of nuclear reactors. We report the modification of an existing 252Cf-based instrument and construction of a shielded, high-efficiency counting system for the in vivo measurement of Al in the hand. Phantoms containing tissue-equivalent solutions of Ca, P, Na and Cl with various Al loadings were used for validation of the technique. The Al/Ca ratio in the hands of seven patients with renal failure was measured using a cyclic activation technique to compensate for the relatively low neutron output of the 252Cf source, and a detection limit of approximately 2.2 mg Al was achieved for a dose equivalent of 36 mSv. The results were compared with the Al content of iliac crest bone biopsy specimens measured using electrothermal atomic absorption spectrophotometry. PMID- 8401273 TI - Continuous meconium monitoring during labour using an intrauterine probe. AB - The fetus discharges meconium (its bowel contents) into the amniotic fluid during labour in approximately 10% of pregnancies. In about 10% of cases where meconium is passed, the fetus gasps, inhaling the sticky meconium into the upper respiratory tract. After birth, the meconium blocks the air passages in the lungs, impairing gas exchange (meconium aspiration syndrome, MAS). Up to 20% of infants suffering from MAS die and recently published studies have shown a long term effect of MAS in causing cough and wheeze. At present, meconium is only noticed at birth or occasionally when amniotic fluid leaks past the presenting part of the fetus. We have developed a system to monitor meconium continuously during labour, using a flexible intrauterine probe. The system provides a measurement of the meconium concentration of amniotic fluid during labour every 2 min, with a 60% prediction interval of +/- 10 g l-1, and a 99% prediction interval of +/- 30 g l-1 (clinically 'thick meconium' contains around 100 g l-1). The noise of the measurement is of the order of +/- 10 g l-1, and the response to changes in the meconium concentration is from 40 to 120 s, depending on its configuration. The system also provides other obstetric variables such as fetal heart rate and uterine activity measurement, obtained from a fetal monitor. Preliminary clinical results indicate that this system can measure meconium not apparent to the attending staff; the system can detect changes in the meconium concentration of amniotic fluid; the measurements are confirmed by visual observation at delivery; and changes of meconium concentration seem to correlate with known stressful stimuli. The system therefore provides a new tool from which new variables are obtained, which can greatly enhance clinical research into the pathophysiology of meconium passage and aspiration. PMID- 8401274 TI - The use of a microcomputer to automate measurement of action potential duration for both transmembrane and monophasic action potentials. AB - Measurement of action potential duration is made more valuable if it can be made simultaneously with other variables, to which it may be related. We have developed a microcomputer-based system which allows measurement of action potential duration, both for transmembrane action potentials and for monophasic action potentials. The system allows simultaneous recording and analysis of action potentials and intraventricular pressures. Both end-diastolic and maximum systolic pressures have been analysed. Action potential duration was assessed at four different levels of the repolarization curve. We have analysed the consistency of measurements made by the computer, and compared them to measurements made manually, using results from six dog experiments. For action potential duration, there was no systematic difference between the manual and the computer methods, but the computer was significantly more consistent. In the case of the pressure measurements, the two methods were approximately the same in their consistency, and again there was no systematic difference. We have demonstrated that potential errors in determination of the average diastolic potential did not significantly affect the results obtained by our method. The variances of action potential duration measurements made at different levels of repolarization were equal. We demonstrated that there was no effect of amplitude on the action potential duration of potentials recorded under steady-state conditions. PMID- 8401275 TI - Air-fluidized beds and their ability to distribute interface pressures generated between the subject and the bed surface. AB - Pressures were measured under five anatomical sites prone to pressures sores for ten subjects, supine and sitting on two different air-fluidized beds. The beds were the Clinitron (trademark, SSI) and the Fluidair Plus (trademark, KCI Mediscus). Mean supine pressures were less than 4 kPa under four sites. The average supine buttock pressure was 2.65 kPa. This increased to 3.71 kPa upon sitting up, though pressures did not rise above the accepted capillary closing pressure, on either bed. Low interface pressures at these sites were due to good moulding between subject and bed. Heel pressures averaging 7.08 kPa, were a factor of 2.67 times greater than buttock pressure, and were higher than expected considering the depth the heels sunk to in both beds. This exceeded the accepted capillary closing pressure and was attributed to covering sheets preventing true floatation at the heels. PMID- 8401276 TI - A transducer for detecting foetal breathing movements using PVDF film. AB - Foetal breathing movement (FBM) in utero has come to play an important role in foetal diagnosis. FBM may be monitored using real time ultrasound imaging of the foetus in utero. Foetal breathing activities can also be detected by monitoring maternal abdominal wall movement in the frequency range of 0.5-2.5 Hz. This paper presents a transducer which detects FBM non-invasively by monitoring maternal abdominal wall movements. Foetal heart sounds can also be monitored. The transducer presented uses piezo-electric film as the transducing medium. Results from preliminary clinical trials of prototype transducers on 10 patients are discussed. PMID- 8401277 TI - Malfunction of medical equipment as a result of mains borne interference. AB - Medical equipment has become more intelligent as the manufacturers have incorporated the latest microprocessor based technology. Equipment malfunction can be caused at any time by inherent errors in the control program but it is particularly important that this is designed to cope with the effects of electrical interference which, in addition, may cause corruption of the software. We have considered interference found in the mains supply in the hospital environment. Using a test protocol with appropriate interference simulators, a wide range of medical equipment was removed temporarily from use and its immunity to electrical mains borne interference tested. Battery operated mains rechargeable devices were unaffected by mains voltage variations including drop outs and sags whereas mains powered devices were affected to varying degrees of severity. In particular, repetitive drop-outs caused loss of power due to fuse blowing in some life support equipment. Impulses affected 25% and pulse bursts 50% of the equipment tested with some evidence that the more recent designs coped better. The EEC Directive on electro-medical compatibility compliance may cause the design of equipment to be improved but hospitals will have to cope with the above problems in their existing equipment for many years to come. PMID- 8401278 TI - Digital audio tape as a method of storing Doppler ultrasound signals. AB - Digital audio tape recorders perform significantly better than conventional tape recorders in respect of their frequency response, dynamic range, signal-to-noise ratio, and wow and flutter figures. They also have advantages in terms of their tape size and playing time, and their ability to record additional information as 'sub-codes'. The suitability of DAT recorders as a means for storing ultrasound Doppler signals has been evaluated. The results show that they are an ideal instrument for recording directional Doppler signals, but like conventional tape recorders are not suitable for storing quadrature Doppler signals. PMID- 8401279 TI - Measurement of forces applied during the clinical cementation of dental crowns. AB - Uncertainty exists about the forces applied by dentists during dental crown cementation. A measuring system was developed based around a commercially available miniature (3.8 mm high and 12.7 mm diameter) load cell. The load cell was mounted in a finger stall and the applied force measured. Experimental results suggest that dentists typically apply a force to metal crowns of about 60 N for a few seconds, followed by the application of a steady force of about 20 to 30 N. Lower forces are applied to porcelain crowns. PMID- 8401280 TI - Within-breath measurement of respiratory impedance. AB - The measurement of the complex respiratory impedance at a high temporal resolution is described. The measurement of impedance is correlated with the breathing cycle using electrical transthoracic impedance. This allows the determination of the complex impedance at any fixed point in the respiratory cycle, which results in lower variability in the measured data when compared to the original device. The respiratory impedance in normal subjects fluctuates during the breathing cycle and has two maxima during a single breath, which occur at peak inspiratory and expiratory flow. In addition, the measurement of impedance at a high temporal resolution enables the observation of novel features in the impedance which appear to be associated with certain respiratory disorders. PMID- 8401281 TI - [Genetic polymorphism of the human genome locus containing the tumor necrosis factor gene: ethnic differences in the allele frequency distribution]. PMID- 8401282 TI - [The role of N and E GTP-binding centers of tubulin during its assembly into microtubules]. PMID- 8401283 TI - [A new antagonist of the physiological effects of the learning stimulant ACTH(4 7)]. PMID- 8401284 TI - [The role of sialic acids in polymorphism of sturgeon gonadotropin]. PMID- 8401285 TI - [Ultrastructure of rat muscle spindle pole under common and load conditions]. PMID- 8401286 TI - [Assessment of virus resistance of transgenic tobacco and alfalfa plants bearing the human beta-interferon gene]. PMID- 8401287 TI - The biased lamellipodium development and microtubule organizing center position in vascular endothelial cells migrating under the influence of fluid flow. AB - To analytically study the morphological responses of vascular endothelial cells (ECs) to fluid flow, we designed a parallel plate flow culture chamber in which cells were cultured under fluid shear stress ranging from 0.01 to 2.0 Pa for several days. Via a viewing window of the chamber, EC responses to known levels of fluid shear stress were monitored either by direct observations or by a video enhanced time-lapse microscopy. Among the responses of cultured ECs to flow, morphological responses take from hours to days to be fully expressed, except for the fluid shear stress-dependent motility pattern change we reported earlier which could be detected within 30 min of flow changes. We report here that ECs exposed to more than 1.0 Pa of fluid shear stress have developed lamellipodia in the direction of flow in 10 min. This is the fastest structurally identifiable EC response to fluid shear stress. This was a reversible response. When the flow was stopped or reduced to the level which exerted less than 0.1 Pa of fluid shear stress, the biased lamellipodium development was lost within several minutes. The microtubule organizing center was located posterior to the nucleus in ECs under the influence of flow. However, this position was established only in ECs responding to fluid shear stress for longer than 1 h, indicating that positioning of the microtubule organizing center was not the reason for, but rather the result of, the biased lamellipodium response. Colcemid-treated ECs responded normally to flow, indicating that microtubules were not involved in both flow sensing and the flow-induced, biased lamellipodium development. PMID- 8401288 TI - The spatial distribution of spindle microtubules in anaphase 3Y1 cells. AB - The spatial distribution of the microtubules (MT) in the rat 3Y1 cells in mitosis was investigated by immunoelectron microscopy and computer-graphic reconstruction of serial thin sections. In anaphase the interzone-MT increased in number gradually with advancing phase, while the kinetochore-MT in half-spindles decreased. The interzone-MT overlapped with each other at the equatorial region of the cell, and they formed a specific structure called the 'stem bodies'. The ends of the interzone-MT opposite to the stem bodies often attached to chromosomes but not to the poles. The stem bodies were not labeled with immunogold particles of anti-alpha tubulin. Some of the stem bodies or MT which originate from stem bodies were found just beneath the plasma membrane in the equatorial region where abundant actin filaments appear showing the formation of the contractile ring and subsequently the cleavage furrow begins. On the basis of these observations it is assumed that the interzone-MT is involved both in the separation of chromosomes in anaphase and in the formation of the cleavage furrow in telophase. PMID- 8401289 TI - Enhancement of phorbol ester induced cell aggregation after alterations in asparagine-linked oligosaccharides. AB - Human erythroleukemia (K-562) cells grown in the presence of phorbol 12,13 dibutyrate formed aggregates of cells not seen in untreated control cultures. Furthermore, the proportion of cells in aggregates and the size of the aggregates both increased dramatically in cultures treated with both phorbol ester and kifunensine, an inhibitor of asparagine-linked oligosaccharide processing. Relative to control cells, phorbol ester treated cells exhibited a greater proportion of N-linked oligosaccharides of the complex-type. Kifunensine prevented this change and caused an accumulation of Man9GlcNAc2. The enhanced aggregation of cells treated with phorbol ester plus kifunensine depended on phorbol ester concentration and was blocked by inhibitors of protein kinase C (H7, sphinganine and sangivamycin). In flow cytometry analysis, phorbol ester treated K-562 cells showed an increase in CD44, a glycoprotein involved in cell adhesion. Moreover, monoclonal antibody to CD44 augmented reaggregation of phorbol ester treated cells. The results implicate phorbol ester induction of CD44 in aggregation of K-562 cells and demonstrate that the presence of high mannose-type asparagine-linked oligosaccharides on cell glycoproteins correlates with increased aggregation of phorbol ester treated cells. PMID- 8401290 TI - Bistramide A-induced irreversible arrest of cell proliferation in a non-small cell bronchopulmonary carcinoma is similar to induction of terminal maturation. AB - Bistramide A, a new toxin isolated from a New Caledonian Urochordata, shows an antiproliferative effect on a non-small-cell lung carcinoma line in vitro and G1 blockade. In this work, the growth arrest induced by bistramide A was shown to be irreversible as assessed by growth kinetics of pretreated cells. Furthermore, the drug caused an underexpression of the nuclear antigen Ki67. These events are similar to a G1-differentiation cell cycle step blockage and a terminal maturation induction. PMID- 8401291 TI - Lithium induces changes in the plasma membrane protein pattern of early amphibian embryos. AB - The plasma membrane protein pattern of Rana ridibunda embryos subjected to lithium (Li) treatment at various stages of development was examined by two dimensional gel electrophoresis. Differences were observed at the neurula stage not only as compared to controls but among lithium-treated embryos as well. Of particular interest was the presence of proteins, specific for the gastrula stage, in lithium-treated embryos. The results are discussed in relation to the well-known effect of lithium on amphibian morphogenesis. PMID- 8401292 TI - Characterization of the non-sulphated highly anionic Kurloff cell glycoconjugates by lectin- and immunoblotting: evidence for the presence of alpha (2,8)-linked polysialic acid-bearing molecules. AB - Urea or guanidine hydrochloride-soluble extracts from highly purified Kurloff cells (KC) radiolabelled in vitro were subjected to DEAE-cellulose chromatography. Among the three anionic peaks obtained, a major and non-sulphated peak (designated as peak IV) strongly affected by glucosamine-labelling and eluted at about 0.3 M NaCl was analyzed. Gel filtration on Sepharose CL4B and 10% SDS-PAGE indicated its heterogeneous size. Peak IV consisted mainly of N-glycans as shown by its susceptibility to tunicamycin. Further insight into its chemical nature was obtained by examining its binding capacity to different lectins and by immunodot analysis. It strongly interacted with concanavalin A (Con A) after dot blot or Western blotting. A large amount of these glycoproteins is not of the high-mannose type since Galanthus nivalis agglutinin reacted weakly with peak IV. Moreover, bindings to Phaseolus vulgaris and to wheat germ agglutinins suggest the presence of bisecting N-acetylglucosamine residues. Bindings to Sambucus nigra and to Ricinus communis agglutinins, dramatically lessened and increased respectively after desialylation, suggest the presence of Neu5Ac alpha 2,6Gal/GalNAc sequences. The absence of outer sialic acid residues linked alpha 2,3 to galactose was demonstrated following Maackia amurensis agglutinin negativity. The use of poly(alpha 2,8-sialyl) endo-N-acylneuraminidase combined with immunodot procedure with a monoclonal antibody that specifically recognizes alpha 2,8-linked polysialic chains revealed that peak IV contains oligosaccharidic epitopes common to polysialylated neural cell adhesion molecules. PMID- 8401293 TI - The structural organization of the pathogenic protozoan Tritrichomonas foetus as seen in replicas of quick frozen, freeze-fractured and deep etched cells. AB - The quick-freezing and freeze-etching technique was used to analyse the cytoskeleton of Tritrichomonas foetus, a pathogenic protozoan of the urogenital tract of cattle. The cytoplasm presented a network of filamentous structures interacting with each other, with the surface of the hydrogenosomes and the nuclear membrane. Two nm wide filamentous structures were found in the luminal space of the Golgi complex, connecting the two faces of each cisterna. The microtubules of the pelta-axostyle system were connected by bridges 30-40 nm long and 10 nm wide, regularly spaced with an interval of 25 nm. The costa is a structure formed by a complex array of filaments and globous structures. It seems to be connected to the recurrent flagellum through a complex network formed by 15 and 10 nm wide filaments which emerge from the peripheral region of the costa and penetrate into the surface projections of the protozoan body to which the recurrent flagellum is attached. Other filaments were seen connecting the surface of these projections with the surface of the flagellum. PMID- 8401294 TI - Immunodetection of low concentrations of ovalbumin in cryoprocessed quail oviduct cells by semi-automatic quantitative analysis. AB - In a previous paper (Quintana et al, Biol Cel 72, (1991) 167-180) we reported the anti-ovalbumin antibody immunolabeling in the cytoplasm of tubular gland cells from cryofixed, cryosubstituted and acrylic-resin embedded quail oviduct. To confirm these preliminary visual observations, we have carried out a semi automatic quantitative study of immunogold labeling. The quantitative results confirm the previous data. In addition, we found a significant higher immunolabeling with low temperature Lowicryl K11M embedding procedure as compared with high temperature LR White embedding. PMID- 8401295 TI - Association of spectrin with manchette microtubules in mammalian spermatids. AB - In hamster and mouse spermatozoa a spectrin immunogold labeling was found under the plasma membrane in the principal piece of the flagellum. During spermatid differentiation, the spectrin labeling was associated with the manchette, a transient microtubular network involved in nuclear shaping and organelle translocation. PMID- 8401296 TI - FAD-glycerophosphate dehydrogenase activity in pancreatic islets and liver of ob/ob mice. AB - The activity of FAD-glycerophosphate dehydrogenase, as measured through the generation of either 3HOH from L-[2-3H]glycerol-3-phosphate in the presence of FAD or iodoformazan from iodonitrotetrazolium, displayed comparable values in islet homogenates of lean and obese (ob/ob) mice. In the liver of the obese animals, the results obtained by the colorimetric and radioisotopic assays yielded a paired ratio twice higher than in control mice. Although isoforms of the mitochondrial enzyme could be present in variable proportions depending on the cell type and genetic background, the present results suggest that, in ob/ob mice, the increased secretory responsiveness of the islet B-cell to D-glucose coincides with an unaltered activity of FAD-glycerophosphate dehydrogenase. This contrasts with the situation recently documented in db/db mice, in which an impaired secretory response of the B-cell to D-glucose is associated with a decreased activity of FAD-glycerophosphate dehydrogenase. PMID- 8401297 TI - Enhanced c-fos expression after intracellular iron deprivation. AB - Intracellular iron deprivation by deferoxamine treatment, which leads to cells arrest in the S phase, enhanced c-fos expression in the neuroblastoma cell line, IMR32. The c-fos expression of iron deprived cells retained its response to stimulation by TPA, and cytosolic PKC activity did not decline after iron deprivation. The data suggest that PKC was not down-regulated. Creatine kinase activity also remained constant in the cytosol of iron deprived cells, indicating intact cellular function. Iron deprivation may activate the growth-related oncogene, c-fos, through some means other than the PKC pathway. PMID- 8401298 TI - Role of lipid peroxidation in impairment of mitochondrial function at complex I by methyl isocyanate treatment of rats in vivo. AB - The subcutaneous administration of methyl isocyanate (MIC) in 1.0 LD50 dose in rats caused a significant effect on hepatic mitochondrial function only at complex I region of the respiratory chain. MIC administration at 1.0 LD50 dose also resulted in significant increases in malondialdehyde and ferrous ion concentration in liver mitochondria. It is suggested that the augmented lipid peroxidation in hepatic mitochondria, catalyzed by iron, possibly mobilized from intracellular stores leads to the inhibition of enzymes of mitochondrial respiration at complex I region, in vivo, in rats receiving a lethal dose of MIC subcutaneously. PMID- 8401299 TI - Small angle X-ray scattering study of S100 proteins. AB - Small-angle X-ray scattering of both S100b and a mixture of S100a and S100ao (S100a,ao) was measured over a protein concentration range of 1.5 to 10.0 mg/ml. The addition of trifluoroperazine (TFP) to S100 solutions suppressed higher aggregation under the conditions studied. In the presence of TFP (1 mol TFP/mol of protein dimer), the radius of gyration of S100b and S100a,ao is found to be 19.5 +/- 0.3A and 20.2 +/- 0.3A, respectively, indicating that most S100 proteins may exist as dimers under the conditions studied. The observed difference (0.7 +/ 0.3A) in the radius of gyration between S100b and S100a,ao indicates that these dimers have different, asymmetrical shapes. PMID- 8401300 TI - Globin chain synthesis in hemolytic anemia reticulocytes. A case of hemoglobin Burke. AB - Peripheral red blood cells from an anemic patient were incubated with [14C]Leu and the labeled globin chains were analyzed by CM-cellulose column chromatography in 8M urea. The radioactivity almost equal to that of beta chain emerged between beta and alpha chains. The newly appeared materials were found to be derived from an abnormal hemoglobin (Hb), Hb Burke [beta 107(G9)Gly to Arg], by the sequence analyses of abnormal globin chain and beta globin mRNA from the patient. PMID- 8401301 TI - Isolation and characterization of a novel microsomal membrane-bound phenol sulfotransferase from bovine liver. AB - A novel phenol sulfotransferase (PST) was detected in bovine liver microsomal membrane fraction. The enzyme was found to be capable of catalyzing the sulfation of simple phenolic compounds, with 3'-phosphoadenosine-5'-phosphosulfate as the sulfate donor. Detergent extracted PST showed a pH optimum of 5.7 and, among the simple phenols tested, the PST exhibited highest activity toward alpha-naphthol. No activities were detected when tyrosine and its derivatives were used as substrates. Both 2,6-dichloro-4-nitrophenol and chlorpromazine were capable of inhibiting the activity of the PST toward p-nitrophenol with inhibition Coefficient50 values of 100 nM and 4 mM, respectively. PMID- 8401302 TI - Molecular cloning and characterization of PstI fragments of bovine herpesvirus-1 DNA. AB - The DNA from an Indian isolate of BHV-1 was cleaved with the restriction enzyme PstI and cloned into pUC9 plasmid. Two recombinant plasmids were then analyzed using restriction endonucleases PstI, EcoRI, BamHI, SmaI, SalI, HindIII, BglI, HaeIII, XhoI, RsaI and StuI. On the basis of digested fragments of recombinant plasmids, two plasmids could be mapped for restriction enzymes SalI, StuI, SmaI and XhoI using single and double digestion. PMID- 8401303 TI - Glycogen-bound protein in lower eukaryote and prokaryote. AB - The proteoglycogen fraction of Neurospora crassa was purified and subjected to radioiodination with [125I]iodide. Amylolysis of the polysaccharide moiety led to the isolation of a labelled 31 kDa-protein. The NH2-terminal amino acid sequence of 10 residues of the 31 kDa-protein was determined. A 31 kDa-protein was also bound to glycogen in Escherichia coli. Proteoglycogen has not been heretofore found in any primitive unicellular organism. PMID- 8401304 TI - On the two pathways of the M-intermediate formation in the photocycle of bacteriorhodopsin. AB - The flash-photolysis technique was used to study the photocycles of the wild-type bacteriorhodopsin (WT bR) and D96N mutant. Kinetics of the L-intermediate decay and M-intermediate formation at pH 7.0, 20 degrees C fit well a sum of two components having time constants, tau (1) = 60 microS and tau (2) = 250 microS, for the WT bR, and a sum of three components having time constants, tau (1) = 55 microS, tau (2) = 220 microS and tau (3) = 1 mS, for the D96N mutant. The fast component with a time constant of 1.4 microS was found in the photoresponse at 400 nm. It constituted 10% of the total amplitude and may be attributed to the K- >L transition. The component with tau = 1 mS was observed in the photocycle of the WT bR as a lag phase in the relaxation of the photoresponse at 400 nm. The difference absorbance minima, corresponding to the first (55-60 microS) and the second (220-260 microS) components of the M-formation, were located at 550 and 530 nm, respectively. The absorbance spectra, corresponding to the 1-mS-component of the M-formation of the D96N bR, may be represented as a superposition of spectra, corresponding to the first and the second components in the region of 460-700 nm. The effect of azide on the D96N bR revealed two azide-independent components in the decay of L-intermediate. Azide was shown to protonate all M forms simultaneously. This indicates that the Schiff base pK rises almost immediately after deprotonation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401305 TI - Immunomodulation and 'Ia' variability in dendritic lymph cells: a radiobinding immunoassay versus immune response study. AB - A radiobinding immunoassay technique has been employed to determine the 'Ia' antigens localised on the surface of the following immunocompetent cells: Dendritic lymph cells from peripheral intestinal lymph, peritoneal macrophages and thymocytes all gathered from sheep red blood cell treated animals, and the irradiated cell controls. A corresponding popliteal lymph node assay was performed following the administration of the same cells into the foot pad of allogeneic rats to determine the comparative immunogenic responsiveness. Dendritic lymph cells in free flowing lymph of sensitised, irradiated and control animals showed a significantly higher 'Ia' content and consequently greater immunogenicity as measured by the popliteal lymph node assay. Sheep red blood administration in animals may stimulate surface 'Ia' expression in lymph derived dendritic cells resulting in higher immune-responsiveness. PMID- 8401306 TI - A gene for cytochrome c oxidase subunit II in duck mitochondrial DNA: structural features and sequence evolution. AB - The gene encoding cytochrome c oxidase subunit II (COXII) was amplified by polymerase chain reaction (PCR) using mitochondrial DNAs (mtDNAs) isolated from two genera of domesticated ducks, Anas platyrhnchos and Cairina muschata, and their intergeneric and intrageneric hybrids as templates. Comparison of the nucleotide sequences from the two different duck lineages shows that they have approximately 91% homology. The nucleotide substitution pattern in the duck COXII gene reveals a high transition/transversion ratio (8.8:1.0) while 93.2% of the nucleotide substitutions are silent. The duck COXII, which uses GUG as the initiation codon, contains 228 amino acid residues. Comparison of the deduced COXII amino acid sequences in duck, chicken, Xenopus laevis, mouse, bovine and human reveals five highly conserved domains. The evolutionary conservation of the primary structure of COXII implies its functional importance. PMID- 8401307 TI - The use of trehalose-stabilized lyophilized methanol dehydrogenase from Hyphomicrobium X for the detection of methanol. AB - The enzyme methanol dehydrogenase (EC 1.1.99.8) from Hyphomicrobium X was used in an attempt to develop a rapid colorimetric test for methanol. The enzyme was stabilized for storage by lyophilization in the presence of the disaccharide trehalose. It was found that the enzyme retained significantly greater activity in the dried state with trehalose than without. The enzyme was partially purified by ammonium sulphate fractionation, after which it was found to be more stable in solution at pH 9 than at pH 7. A procedure is given which involves mixing a defined amount of enzyme with the methanol-containing water together with phenazine methosulphate (PMS), 2-6-dichlorophenol-indophenol (DCPIP) and cyanide, and observing the resultant colour change from blue to yellow if methanol is present. The sensitivity of the procedure is such that 9 mg L-1 of methanol can be readily detected. PMID- 8401308 TI - Expression of oryzacystatin cDNA using yeast artificial chromosome under ADH promoter in baker's yeast. AB - We constructed an expression vector into a yeast artificial chromosome (YAC) harboring the cDNA for oryzacystatin, a cysteine proteinase inhibitor from rice, under the control of the yeast ADH promoter. When the expression vector was introduced into Saccharomyces cerevisiae in the form of either an artificial chromosome or a circular plasmid, transformants carrying the DNA grew well in a selective medium. However, the content of the introduced DNA decreased significantly during passages in non-selective YPD medium. The stability of the introduced DNA was enhanced in selected clones obtained as colonies viable in selective medium after many passages in YPD medium. The stable transformants thus obtained expressed the mRNA for oryzacystatin at levels as high as those of intrinsic yeast ADH. PMID- 8401309 TI - Isolation of intestinal cell peroxisomes by rate-dependent banding in a vertical rotor. AB - The preparation of peroxisome-enriched fractions from mouse intestinal epithelial cells is possible by ultracentrifugation using the vertical rotor under rate zonal conditions. The run time is short which results in a low total centrifugal effect to which these fragile organelles are subjected. The banding of peroxisomes occurs in the central region of the sucrose gradient whereas the mitochondria band in the lower regions. Good separation from the mitochondria is achieved with contaminants resulting from brush-border fragments and endoplasmic reticulum. Comparisons are made with preparations using a swing-out rotor and a zonal rotor. Electron microscope examination of the preparations confirms the presence of diaminobenzidine-positive particles which resemble the peroxisomes seen in intact cells. In addition, it has been shown that density gradients can be constructed which prevent large particles from impacting and causing wall effects. PMID- 8401310 TI - Demonstration of putative subunit(s) of a novel insulin receptor-associated 66 kDa GTP-binding protein, Gir. AB - A 66 kDa GTP-binding protein, Gir, and insulin receptor (IR) were copurified from human placental membrane by DEAE-Sephacel and Wheat Germ Agglutinin (WGA) Sepharose affinity chromatography. The WGA-fraction containing IR and Gir (IR-Gir fraction) was phosphorylated (95 kDa IR-beta and 66 kDa Gir) by IR-tyrosine kinase using [32P]ATP or photolabeled with [32P]8-azido-GTP (mainly 66 kDa), and was cross-linked with a bifunctional reagent, bis-[sulfosuccinimidyl] suberate (BS3). The Gir cross-linked with putative subunit(s) to form a 110 kDa complex. Phosphorylation as well as 8-azido-GTP binding to Gir was not affected by cross linking indicating that like other G-Proteins, Gir may also have subunits and that cross-linking of Gir with its putative subunits(s) does not block GTP binding and phosphorylation sites. PMID- 8401311 TI - Purification and properties of murine corneal aldehyde dehydrogenase. AB - Murine corneal aldehyde dehydrogenase has been purified to homogeneity and characterized with a range of aldehyde substrates at pH 7.4. The enzyme was a dimer with a subunit molecular weight of 59 KDa. and appears to prefer aldehyde products of lipid peroxidation as substrates. The enzyme constituted approximately 5% of the total soluble protein of mouse cornea. A dual role has been proposed for corneal aldehyde dehydrogenase in providing the eye with protection against UV-B light: by oxidizing aldehydes generated through light induced lipid peroxidation; and by the direct absorption of UV-B light by the enzyme. PMID- 8401312 TI - Polyamines in Trypanosoma cruzi. AB - Growth and polyamine content of epimastigotes of Trypanosoma cruzi were studied in the presence of precursors or inhibitors of putrescine synthesis. Arginine and agmatine turned out to be better precursors than ornithine. alpha-D difluoromethylarginine, an inhibitor of arginine decarboxylase, inhibited cellular proliferation and decreased putrescine, spermidine and spermine contents while that of cadaverine remained unchanged. These effects were reversed both by agmatine and putrescine. alpha-D-difluoromethylornithine, an inhibitor of ornithine decarboxylase did not inhibit growth of parasites grown in a polyamine free medium. These results suggest that epimastigotes need polyamines to grow, and that the parasite are able to synthesize them mainly through the arginine decarboxylase pathway. PMID- 8401313 TI - Phenotypic and cytogenetic characterization of a human corpus cavernosum cell line (DS-1). AB - We report here the establishment and characterization of a corpus cavernosum cell line (DS-1) from human penile tissue. This is the first cell line of its type derived from cavernosum tissue. DS-1 cells have become immortalized in culture, and show growth in monolayers. These cells have a doubling time of about 45 h in in vitro culture. Cytogenetic analysis by G-banding demonstrated a diploid karyotype with a model chromosome number of 46. The chromosome constitution of DS 1 cells was found to be male (XY), in 28/30 cells scored. Two of the 30 cells showed an extra structurally rearranged "marker" chromosome, that appeared to be a derivative of chromosome 18 with excessive chromosome on the short arm. Ploidy analysis revealed that the majority of DS-1 cells had a DNA index of one. About 35% cells were found to be in G-1 phase and 52% cells in S phase. Light and electron microscopy of DS-1 cells and original penile tissue showed typical characteristics of this tissue. Immunocytochemistry studies using antibodies to smooth muscle actin, desmin, vimentin and cytokeratin (LP34, CAM5.2) showed that the DS-1 cell line had predominantly smooth muscle cells, as these cells were positive for smooth muscle actin, desmin and vimentin. PMID- 8401314 TI - Cloning of the BamHI repeat from Amaranthus and study of its methylation in genomic DNA during dedifferentiation. AB - A 670 bp BamHI repeat was isolated from the genomic DNA of Amaranthus paniculatus and was cloned in pUC19 vector (pABH3). The level of methylation of total genomic DNA and the BamHI repeat therein was studied by employing methylation sensitive enzymes to digest genomic DNA isolated from whole seedlings, hypocotyl, callus, and suspension culture of Amaranthus. The restriction pattern indicated double methylation of the target site CCGG. Methylation pattern of both the total genomic DNA and the cloned repeat was found to be similar during dedifferentiation indicating that there may not be a general relationship between hypomethylation and dedifferentiation. PMID- 8401315 TI - Activation of a heparin-degrading enzyme by a 'protein matrix' effect. AB - An unusual activating effect of protein on Flavobacterium heparinase is described. The phenomenon is nonselective with respect to protein species, but does not occur with other biomolecules such as nucleic acids, polysaccharides, or free amino acids. We show that protein activates heparinase over broad ranges of temperature and ionic strength, and stabilizes the enzyme against both reversible and irreversible structural changes. The nonselective activation of an inducible enzyme by protein may be an important regulatory mechanism in microenvironments in which the concentration of organic material may vary. PMID- 8401316 TI - Characterization of trypsin inhibitors from Tora-mame seeds, one of the Japanese cultivars of Phaseolus vulgaris. AB - Four isoinhibitors against bovine pancreatic trypsin were purified from Phaseolus vulgaris(cv. Tora-mame) seeds by extraction with water(pH 2.0), ammonium sulfate fractionation, gel chromatography on Sephacryl S-200, trypsin-Sepharose gel affinity chromatography, and chromatofocusing. They inhibit both trypsin and chymotrypsin strongly. Their molecular masses are 85 kDa, estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Their isoelectric points range 5.09 to 4.46. They are high in the content of aspartic acid, serine, proline, and half-cystine but low in valine, methionine, tyrosine, and phenylalanine. Tryptophan is absent from them completely. They are bound to both trypsin and chymotrypsin with equimolar ratio, and have separate and independent binding sites for both proteases. Chemical modification showed that the inhibitors are of lysine type. PMID- 8401317 TI - The ability of apocytochrome C to pass lipid bilayer is relevant with its folding state. AB - The translocation ability of two different species of apocytochrome c(horse heart, Candida krusei) across the soybean phospholipid vesicles decreased in the order: C. krusei > horse heart. Theoretical calculations of the amphiphilicity of their N- and C-terminal alpha-helix formation and the release experiment using calcein-enclosed soybean phospholipid vesicles showed that there was no direct relevance between their import ratio and the amphiphilic helicity. On the other hand, taking the advantage of circular dichroism technique, a more loosely folded structure of C. krusei following the interaction with soybean phospholipid vesicles was observed which should be responsible for the difference in translocation rate. PMID- 8401318 TI - Stimulation of nerve growth factor synthesis/secretion in mouse astroglial cells by coenzymes. AB - We examined the effect of coenzymes such as PQQ, pyrroloquinoline quinone; TOPA, 3-(2,4,5-trihydroxyphenyl)-DL-alanine, and lipoic acid on nerve growth factor (NGF) synthesis in mouse astroglial cells, BALB c/3T3 cells, and WS-1 cells in culture. These coenzymes had a stimulating effect on NGF synthesis without causing cytotoxicity. Especially PQQ showed the strongest activity of promoting NGF synthesis in astroglial cells, whereas lipoic acid had the strongest effect on BALB c/3T3 cells. The activity may not due to the catechol ring or 1,4 benzoquinone ring, but due to the oxidative or reductive activity. These results suggest that these coenzymes may play a role in NGF synthesis and neuronal survival through the stimulating effect of the NGF synthesis in brain and such compounds are good candidates as NGF inducers. PMID- 8401319 TI - Salivary and plasma insulin levels in man. AB - Changes in plasma and salivary immunoreactive insulin were studied in saliva and blood of 9 healthy subjects following either oral glucose or a test meal ingestion at 30 min intervals for up to 3h. The levels of plasma immunoreactive insulin reached maximum at 60 min with a value of 62.77 +/- 6.52 microU/ml after the glucose load and 62.62 +/- 6.52 microU/ml after the test meal. In the case of salivary immunoreactive insulin levels, the maximum was attained at 90 min with a value of 18.47 +/- 2.68 microU/min after the glucose load and 22.40 +/- 2.01 microU/min after the test meal. Positive linear correlation (r = 0.74) was found between the salivary and plasma immunoreactive insulin levels. PMID- 8401320 TI - Pyruvate is a lipid precursor for rat lymphocytes in culture: evidence for a lipid exporting capacity. AB - Since acetyl-CoA produced through pyruvate dehydrogenase reaction is poorly oxidized by the Krebs cycle in rat lymphocytes, the fate of acetyl units was investigated in these cells. The results presented here show that 24-h cultured lymphocytes actively synthesize lipids from [3-14C]pyruvate. Furthermore, a considerable amount of these lipids have shown to be exported into the culture medium. Experiments with [1-14C] acetate as a lipid precursor showed a close similarity with the rates of incorporation of [3-14C] pyruvate into the same lipid fractions. Treatment of lymphocytes with the mitogen concanavalin A (Con A) markedly enhanced [1-14C] acetate incorporation into a variety of lipids, but the lectin did not affect [3-14C] pyruvate incorporation. The results suggest that lymphocytes convert pyruvate into lipids via the acetyl-CoA pathway and that Con A interferes in lymphocyte lipogenesis but does not seem to affect the pyruvate dehydrogenase reaction. The ability to incorporate pyruvate into certain lipids may have an important role for the rapidly dividing capacity of lymphocytes since the human cancer strain HeLa 155 (a quickly proliferating cell line) also exhibits this feature by converting much more [3-14C] pyruvate into lipids than do lymphocytes. In addition, comparative experiments with lymphocytes, peritoneal macrophages and HeLa cells indicate that pyruvate may provide precursors for cells with active lipid producing and exporting capacities. PMID- 8401321 TI - Expression of a globin gene in Caenorhabditis elegans. AB - Oligonucleotides related to parts of a globin-like sequence in the genome of Caenorhabditis elegans were used to probe a cDNA library from the same species. A complete globin-like sequence was found in the cDNA, showing that a globin gene appears to be expressed. The hypothetical protein was compatible with the conventional globin fold but may be truncated in the B helix, as in Chironomus globin III. An intron in the codon for residue E3 in the E helix was removed in expression. An initiation codon preceded the globin but the sequence upstream (extending for 30 nucleotides to the vector ligation site) had characteristics both of the code for a protein hydrophobic leader and of a trans-spliced RNA leader. The evidence indicates that C. elegans globin has a single domain, unlike some nematodes that express two tandem globin domains in a continuous translation product, and from its sequence may be predicted to have a high affinity for oxygen. PMID- 8401322 TI - Dual effect of carbachol on the muscarinic receptor. AB - The carbachol-mediated inhibition of cAMP synthesis in membranes of rat cerebral cortex revealed maximum in 0.3 mM carbachol solution, while further increases of the drug concentration blocked the inhibitory effect and restored the adenylate cyclase activity. The bell-shaped dose-response curve was analyzed assuming that carbachol binds with two distinct sites on muscarinic receptor, which are responsible for agonistic and antagonistic effects, related to this receptor. The pKagon and pKant values for carbachol were 5.0 and 2.1, respectively. PMID- 8401323 TI - Effects of insulin on TGF-beta 1-induced cell growth inhibition in the human hepatoma cell lines. AB - The effects of insulin on cell growth control by transforming growth factor beta 1 (TGF-beta 1) in human hepatoma cell lines were studied. TGF-beta 1 inhibited cell growth and DNA synthesis of Hep3B cells but not that of HA22T/VGH cells. The cell cycle-dependent p34cdc2 kinase activity was inhibited by TGF-beta 1 in a dose-dependent manner in Hep3B cells. In contrast, the p34cdc2 kinase activity of HA22T/VGH cells was not regulated by TGF-beta 1. When insulin (10(-7) M) was added simultaneously with TGF-beta 1, we found that the inhibitory effects of TGF beta 1 on cell growth and DNA synthesis in Hep3B cells was completely blocked and the p34cdc2 kinase activity of Hep3B cells was recovered after insulin administration. Thus, cell growth inhibition by TGF-beta 1 in Hep3B hepatoma cells can be antagonized by insulin and their interaction may play an important role in the tumor progression stage of hepatocarcinogenesis. PMID- 8401324 TI - Angiotensin-converting enzyme activator from purified human neutrophils. AB - An endogenous ACE activator has been revealed. Neutrophil-enriched human leucocyte preparations released in isotonic media a relatively thermostable factor, capable of increasing the angiotensin-converting enzyme activity 1.6-2.0 fold. Activator would not pass through a 30 kDa cutoff membrane. Data on the presence of the activator in vivo were obtained in studies of protein-free serum fractions of patients with a hereditary CI-esterase inhibitor deficiency. It was suggested that the neutrophil-released ACE activator could have a role in the modulation of the inflammatory response. PMID- 8401325 TI - Properties of almond beta-glucosidase immobilized on concanavalin A-sepharose. AB - Lyophylized almond beta-glucosidase (EC 3.2.1.21) has been adsorbed onto Concanavalin A-Sepharose (CAS). The yield of the enzymatic units of the CAS enzyme complex preparation was 131%. The values of the kinetic parameters of the free beta-glucosidase were: kM = 1.7 mM and Vmax = 330.1 U/mg protein. The immobilised form showed the following values: kM = 1.7 mM and Vmax = 402.6 U/mg protein. Both enzymatic forms showed essentially the same temperature- and pH activity patterns (temperature optimum: 50 degrees C and pH optimum approx. 6.0), however, the pH stability of the CAS-enzyme complex at pH 6 was significantly higher than the beta-glucosidase in free solution. PMID- 8401326 TI - Comparative analysis of human steroid sulfatase activity in prepubertal and postpubertal males and females. AB - This study analyzes the role of pre or postpubertal stage and sex on the steroid sulfatase activity of human leukocytes. The prepubertal female group (2-7 yrs) presented a higher sulfatase activity than the prepubertal male group (2-7 yrs, 1.99 +/- 0.64 vs 0.99 +/- 0.31 pmol/mg protein, respectively) (p < 0.001), with a female/male ratio of 2.01. The postpubertal subjects (15-40 yrs) showed an activity of 0.77 +/- 0.19 (females) vs 0.56 +2- 0.11 pmol/mg protein (males) and a female/male ratio of 1.37. Enzymatic activity of prepubertal subjects paired by sex was higher than the postpubertal individuals (females p < 0.001 and males p < 0.005). These findings show differences in the steroid sulfatase activity of pre and postpubertal groups suggesting the possible influence of hormones secreted since puberty on the expression of this enzyme. PMID- 8401327 TI - Ornithine decarboxylase inactivation by putrescine: involvement of lipid peroxidation. AB - Effect of polyamines on 12-O-tetradecanoyl phorbol-13-acetate (TPA)-induced reduction of lipid peroxidation was studied. Putrescine protected this lowering of lipid peroxidation in a concentration-dependent manner, but spermidine or spermine could not do so. Putrescine also inhibited the TPA-induced ornithine decarboxylase (ODC) activity and lowered the free sulfhydryl content of TPA exposed mouse skin. These observations indicate that putrescine inactivates ODC probably by lowering SH groups through lipid peroxidation. PMID- 8401328 TI - Some properties of lysosomal arylamidase in rat liver. AB - Lysosomal arylamidase in rat liver was reassessed. The arylamidase was distinguishable into seven types having different isoelectric points by isoelectric fractionation. All the types hydrolyzed valine beta-naphthylamide most rapidly among amino acid beta-naphthylamides tested, but did not hydrolyze arginine beta-naphthylamide. These enzymes exhibited maximum activities at pH 7.0. They had a molecular weight of 135,000 by gel filtration on a Sephacryl S 200. They were all inhibited by sulfhydryl-blocking reagents and activated by sulfhydryl compounds, indicating to be cysteine proteases. PMID- 8401329 TI - Physico-chemical properties of copper-oxidized high density lipoprotein: a fluorescence study. AB - The modifications of the physico-chemical properties of the high density lipoprotein (HDL) before and after in vitro induced oxidation by copper ions have been studied using the fluorescence polarization (Pf) of the phosphatidylcholine derivative of 1,6-diphenyl-1,3,5-hexatriene (DPH-PC) and of the cationic derivative (TMA-DPH). We have observed that HDL oxidation is associated with a decrease of the molecular order at the lipoprotein surface as demonstrated by the increase in Pf with respect to untreated HDL. Moreover in oxidized HDL the polarity-sensitive probe laurdan has shown a decrease of the polarity in its microenvironment. It has been suggested that a decrease in HDL fluidity would inhibit cholesterol reverse transport from peripheral tissues in form of HDL core cholesteryl esters. Peroxidation of HDL, if occurring in vivo, could contribute to the progress of atherogenesis by decreasing cholesterol efflux from peripheral tissues. PMID- 8401330 TI - Novel rat calpastatin mRNA variants. AB - Rat calpastatin cDNAs obtained by RT-PCR method were isolated and directly sequenced; this allowed the identification of two new variants both characterized by a 23 amino acids deletion at the end of the unique N-terminal domain L, increasing the number of functional isoforms. The deletion shows high homology to the amino acid sequence coded by exon 8 of human calpastatin gene and properly conserved splicing consensus sequences, suggesting exon skipping in domain L. Furthermore, three point mutations scattered along the coding region were found. PMID- 8401331 TI - Multiple nuclear protein binding to 135 bp subrepeat element of wheat ribosomal DNA intergenic spacer. AB - Specific binding of wheat nuclear proteins to 135 bp subrepeat element of the ribosomal DNA intergenic spacer was studied by electrophoretic mobility shift assays. A number of specific DNA-protein complexes are readily formed upon incubation of the labelled 135 bp subrepeat element with wheat nuclear protein extracts. Some of these complexes have various DNA-binding specificities as evidenced by their optimal detection in the presence of different non-specific competitor polynucleotides. One of the wheat nuclear proteins was found to bind specifically both to 135 bp subrepeat element and an intergenic spacer region located downstream of the transcription initiation site (between +192 and +366 bp). No apparent effects of the 135 bp element methylation with methylase HpaII on protein binding were found. PMID- 8401332 TI - Effects of arecoline on phase I and phase II drug metabolizing system enzymes, sulfhydryl content and lipid peroxidation in mouse liver. AB - The modifying potential of arecoline alkaloid was studied on hepatic drug metabolizing system enzymes, acid soluble sulfhydryl (-SH) content and microsomal lipid peroxidation. Arecoline was administered intraperitoneally to Swiss albino mice at the dose levels of 10, 20 or 40 mg/kg body wt./day for 10 or 30 days. Significant increase in the levels of glutathione S-transferase (GST), cytochrome b5 (Cyt.b5), cytochrome P-450 (Cyt.P-450) and malondialdehyde (MDA) was observed in the arecoline treated groups. Decreased -SH content was apparent by the administration of 40 mg arecoline for 10 or 30 days. The modulation in biotransformation system enzymes has wide significance in the process of neoplastic development as well as in detecting the further role of biometabolized chemicals. PMID- 8401333 TI - Effects of triiodothyronine on DNA synthesis and differentiation markers of normal human osteoblast-like cells in vitro. AB - To study the direct effects of thyroid hormones on human osteoblasts we examined the effects of triiodothyronine (T3) on proliferation and differentiation of human osteoblast-like (hOB) cells in vitro. T3 increased 3H-thymidine incorporation in DNA of hOB cells (p < 0.05, n = 10). Half maximal effects obtained at a T3 concentration of 1-10 nM which lies within the physiological concentration of the hormone. In addition, T3 increased alkaline phosphatase production (p < 0.05, n = 13) and inhibited procollagen type I carboxyterminal propeptide (PICP) production (p < 0.05, n = 13). T3 interaction with 1,25 dihydroxycholecalciferol (1,25-(OH)2D3) was also studied. 1,25-(OH)2D3 (10(-9)M) alone doubled AP production and induced osteocalcin expression by hOB cells. Concurrent addition of T3 and 1,25-(OH)2D3 did not further increase production of AP, PICP or osteocalcin by hOB cells. In conclusion, T3 exerts significant effects on osteoblast proliferation and differentiation, suggesting that human osteoblasts are targets for thyroid hormones. PMID- 8401334 TI - Determination of halfcystine in proteins as cysteine from reducing hydrolyzates. AB - The separation of cysteine from proline using a variant of the classical cation exchange system for amino acid analysis is described. From reducing hydrolyzates (6 M HCl/0.1% phenol/5% thioglycollic acid, 18 h, 110 degrees C) the halfcystine content of proteins can be accurately determined as cysteine. The system is useful for routine determination of the composition of proteins from a single type of hydrolyzate. PMID- 8401335 TI - [Natural history of bronchial asthma]. PMID- 8401336 TI - [Does the moment of birth have any significance on recognition of atopic diseases of the respiratory system? II. Incidence of atopic diseases in relation to birth date of children from families with increased atopic diathesis]. AB - The aim of this study was to analyze the birth rates in specific quarters of the year in families with a history of atopic diathesis. One hundred families with atopic diathesis with at least two children older than 16 years were studied. The diagnosis of its atopic etiology and demonstration of the specific allergen were all based on results of a questionnaire, physical examination, results of skin tests and serum levels of allergen specific IgE. In the selected 100 families an analysis of birth rates of 251 children was carried out in each quarter of the year and the effect of this on first atopy symptoms was searched for. It was demonstrated that in these families more children were born with atopy (57%) than without. More than half of the children were sensitive to grass pollens and perennial allergens, and only 20% to the perennial allergens. Most children were born during the second quarter of the year. In this group atopy was four fold more often seen. The results demonstrate that the birth date of children from atopic families is a risk factor for atopy. This implies that family planning should be added to the list of secondary prophylactic measures of atopic diseases. PMID- 8401337 TI - [A comparative study on skin tests and allergen-specific IgE levels determined by enzyme allergo-sorbent testing (EAST)]. AB - Ninety seven randomly selected subjects with bronchial asthma and allergic rhinitis were classified for the comparative studies. The level of allergen specific IgE determined by Enzyme Allergo-Sorbent Testing (EAST) was compared with skin tests. The statistically significant correlation was found for 9 selected allergens. However, the Spearman's rank correlation coefficient values varied largely, depending on a given allergen. They ranged from 0.74 (Dermatophagoides pteronyssinus) to 0.33 (Birch pollen) what might be, in the authors' opinion, a result of Bencard allergens use for skin tests and Kallestad allergens for EAST. PMID- 8401338 TI - [Skin tests, total IgE and venom specific IgE and IgG4 in serum of bee-keepers]. AB - In 14 occupationally active bee-keepers, who tolerated stinging well, two diagnostic tests have been performed: 1. skin test at the venom concentration of 10(-3) g/l, 2. both total and venom specific IgE and IgG4 assessments. The study group consisted of 13 male and 1 female aged 34-56 (mean age 44.3). 7 out of 14 bee-keepers (50%) had positive skin tests; mean size being mean = 3.57 +/- 4.07 mm. None of the subjects showed increased total IgE levels, but 71.4% showed bee venom specific IgE (mean mean 1.64 +/- 1.34 FAST). Specific bee venom IgG4 occurred in 13 out 14 individuals (92.8%). Their average serum level was mean 21.74 +/- 17.16 IU/ml. PMID- 8401339 TI - [Behavior of esterase inhibitor C1 in patients with urticaria due to aspirin hypersensitivity]. AB - The aim of the study was to evaluate concentration and activity of C1 esterase inhibitor (C1 INH) in patients with aspirin-sensitive urticaria. Deficiency of C1 INH is the basis for hereditary angioneurotic oedema. The study was performed in 32 subjects with aspirin-sensitive urticaria. The value of C1 INH in examined patients was the same as in control group. It seems there is no coexistence of aspirin-sensitive urticaria and C1 esterase inhibitor deficiency. PMID- 8401340 TI - [Chemotactic activity of serum granulocytes after aspirin in patients with aspirin-sensitive urticaria who find themselves in a state of tolerance for this drug]. AB - The increase in neutrophil chemotactic activity (NCA) is related to degranulation of mast cells. The study included 10 patients in whom aspirin-induced urticaria was related to NCA increase. Tolerance state to ASA was achieved by administering this drug in incremental doses. In none of the examined patients after 600 mg of ASA given during induced tolerance state, the increase of NCA was observed. The authors conclude that in patients with ASA-urticaria, after ASA desensitization, mast cells degranulation does not occur. PMID- 8401341 TI - [Evaluating the safety of bronchoalveolar lavage in patients with bronchial asthma in light of gasometric and spirometric examinations]. AB - Gasometric parameters were performed before, during and after bronchoalveolar lavage (BAL) in 21 patients with mild bronchial asthma. In addition, spirometric parameters were performed before and after BAL. Decrease of pO2, SaO2 and increase of pCO2 which kept within the limits of normal values were observed. After BAL gasometric parameters returned to baseline values. Bronchoalveolar lavage caused statistically significant fall in FEV1 and PEF. BAL was well tolerated and did not worse the clinical state of patients. PMID- 8401342 TI - [Hypersensitivity to weed pollen allergens in the region of Bygdoszcz]. AB - Case records of 8576 patients suffering from hypersensitivity symptoms within upper airways, who were treated at the Otolaryngological Section of the Allergological Outpatient Department of Medical Academy in Bydgoszcz in the years 1975-1989, were analysed. Pollinosis was diagnosed through anamnesis, physical examination and skin tests in 2561 (29.9%) individuals, i.e. 1153 women and 1408 men. Hypersensitivity to weed pollen allergens was found in 1069 (12.5% of all people examined) patients with pollinosis, i.e. 487 women and 582 men. There were 941 (88%) inhabitants of urban and 128 (12%) of rural areas among them. Positive family history of allergic diseases was proved in 616 (57.6%) persons. Mean age at the first visit to the doctor was 28 years for women and 23 years for men, while mean age of weed pollinosis onset was 22 years for women and 17 years for men. The disturbances observed in patients sensitive to weeds were provoked mainly by wormwood (86.2%), mugwort (82.9%), white goosefoot (44.3%), narrow leaved plantain (28.8%), and sheep sorrel (19.0%). Hypersensitivity to grass, tree and/or shrub pollens coexisted in 85.5% of patients with weed pollen allergy and 64.5% of them were also hypersensitive to several non-seasonal allergens in various combinations. Bronchial asthma was recognized in 101 (9.4%) patients with weed pollinosis, while allergic skin reactions were seen in 315 (29.5%) individuals. PMID- 8401343 TI - [Cytograms of nasal mucosa in seasonal and perennial allergic rhinitis]. AB - The role of cytological analysis of nasal mucosa in seasonal and perennial rhinitis was addressed. The study was carried out in 55 subjects (25 females, 30 males) with seasonal allergic rhinitis and 44 patients (28 females, 16 males) with perennial allergic rhinitis. The cytological examination was carried out during the acute phase of the disease and depend on performing a differential count of epithelial and inflammatory effector cells. Patients with perennial allergic rhinitis demonstrated decreased number of cells in comparison with subjects with seasonal rhinitis. The cytogram of these subjects was characterized by increased number of squamous epithelial cells (27.5%) while in seasonal rhinitis the number did not exceed 17.5% (p < 0.05). In addition the number of neutrophils was higher in perennial allergic rhinitis than in pollinosis (respectively 16.8% and 10.6%). Eosinophils were seen more often in seasonal than perennial rhinitis (27.6%, 17.2%, p < 0.05). The increased number of eosinophils correlated with sneezing in pollinosis (r = 0,543, t = 2,186, p < 0.05). This was not demonstrated in patients with perennial allergic rhinitis. The authors conclude that cytograms of nasal mucosa allow to differentiate perennial from seasonal allergic rhinitis. This test is easy to perform and being non-invasive can be used in diagnosis and monitoring. PMID- 8401344 TI - [Analysis of selected spirometric parameters and results of therapy with nedocromil sodium in patients with asthma]. AB - The effect of 12 weeks therapy with nedocromil sodium given as MDI-aerosol, 8 mg/daily in a group of 20 atopic asthmatic patients was studied. The selected spirometric parameters (PEF, FEV1, FVC, FEF25-75%, Raw) and clinical symptoms of bronchial asthma (dyspnoea, cough, sputum) were assessed. Additionally the percentage reduction of bronchodilators usage was obtained. A significant improvement was observed in spirometric parameters. A statistically significant reduction in clinical symptoms of bronchial asthma and bronchodilators usage was noted. PMID- 8401345 TI - [Effect of nedocromil sodium on bronchial reactivity, selected lung function tests and demand for glucocorticosteroid inhalation in patients with bronchial asthma]. AB - In a group of 14 patients with non-atopic asthma the effect on nedocromil sodium (8 mg/day) on bronchial reactivity, lung function parameters and doses of glucocorticosteroid in inhalation was studied. There was the significant reduction in beclomethasone doses, a rise in FEV1, FEF25-75 and decrease of PC20 histamine during 6 weeks of nedocromil therapy. PMID- 8401346 TI - [Bronchodilating action of salbutamol depending on route of administration]. AB - The aim of this study was to demonstrate the differences in salbutamol's bronchodilating effect related to administration routes of the drug. Three different administration routes were assessed: salbutamol given by a MDI device, a nebulizer and intravenously. The following increases of FEV1 were seen after administration routes: MDI--12%, nebuliser--27%, intravenously--22%. These differences were statistically significant. PMID- 8401347 TI - [Clinical analysis of ofloxacin in treatment of postinfectious exacerbations of endogenous bronchial asthma]. AB - The aim of the study was to analyse the clinical effectiveness of ofloxacin in treatment of post-infectious exacerbations of endogenous bronchial asthma. Twenty eight patients (20 males, 8 females) were assessed. The following parameters were evaluated: bacteriological examination of the sputum (before and after treatment), morphological analysis of peripheral blood, bronchoalveolar lavage (performed in 11 patients). Additionally in 7 patients lymphocyte subpopulations of the peripheral blood were determined (OKT4/OKT8). Diurnal variations of serum levels of theophylline were assessed in 12 patients. The authors observed a clinical improvement in 26 patients. Following treatment the white cell blood count (WBC) and the CD4/CD8 ratio decreased. The same observations were made analysing the BAL differential count, in which a marked decrease of neutrophils was seen. The authors conclude that ofloxacin is a safe and efficient antimicrobial agent. PMID- 8401348 TI - [Effect of corticosteroids on tachyphylaxis induced by beta-2- receptor agonists in patients with bronchial asthma]. AB - Fourteen patients with moderate bronchial asthma were selected for the study. Asthmatics were treated with intravenous salbutamol for one week. Peripheral blood lymphocyte beta-2-adrenergic receptor density and affinity were determined before and after the treatment. To investigate an effect of corticosteroids on beta-2-agonist induced desensitization of beta-2-receptors, beta-2-receptor density and affinity following prednisone administration have been determined in lymphocytes in 7 asthmatics. Patients have been given prednisone in the dose of 20 mg daily for 1 wk after salbutamol discontinuation. Spontaneous recovery of beta-2-adrenergic receptor density in the lymphocytes was estimated in further 7 asthmatics, not given corticosteroids following salbutamol therapy. Our data showed that down-regulation of beta-2-adrenergic receptors improves spontaneously and to significantly higher degree after corticosteroid therapy. PMID- 8401349 TI - [N2O cryotherapy in treating allergic rhinitis]. AB - Chronic rhinitis is known since many years to be a difficult therapeutic problem. Also new methods of treatment did not bring satisfactory good results. In our clinic a cryosurgery was applied in patients with chronic rhinitis. Performed observations indicated that cryosurgery was more successful than other conventional methods, especially in early stage of disease. PMID- 8401350 TI - [New drugs in pharmacotherapy of bronchial asthma]. PMID- 8401351 TI - [Potassium channels-opening drugs and their possible use in treating bronchial asthma]. PMID- 8401352 TI - [Historical review of clinical allergology and state of this field in Poland and in the world]. PMID- 8401353 TI - Counterflow centrifugal elutriation: present and future. AB - Counterflow centrifugal elutriation (CCE) has been proposed as a method for separating heterogeneous cell populations into distinct subpopulations on the basis of different sedimentation characteristics, without impairment of cell function or yield. The advantages of this technique are the high recovery and viability of fractionated cells and the rapidity and reproducibility of results. CCE alone or in combination with other separation methods can provide homogeneous populations of cells for further investigations. Recently, CCE has been employed in clinical studies aimed at preventing GVHD in BMT recipients by depleting lymphocytes prior to BM infusion. Furthermore, ongoing studies are concentrating on the use of negative selection procedures on the fractions currently excluded from the graft with the purpose of adding these depleted fractions to the graft preparation to augment the number of stem cells, accessory cells and unselected lymphocyte subsets. In the experimental field, CCE combined with negative and positive selection techniques may be useful in the study of hematopoiesis by separating 'pure' stem cell populations from more committed hematopoietic progenitors. We review here the present and possible future applications of elutriation in the clinical and experimental field. PMID- 8401354 TI - Optimization of conditioning therapy for leukemia prior to BMT. I. Optimal synergism between cyclophosphamide and total body irradiation for eradication of murine B cell leukemia (BCL1). AB - The doses of chemotherapy and radiotherapy and the sequence-dependent influence of CY administration and total body irradiation (TBI) on the eradication of murine B cell leukemia (BCL1) were investigated. Tumor-bearing BALB/c mice were treated with either CY followed by TBI (CY/TBI) or in the reverse (TBI/CY) at different time intervals after injection of 10(6) BCL1 cells. The presence of residual tumor cells after cytoreduction was assessed in secondary BALB/c recipients by monitoring leukemia-free survival. The anti-leukemic effect of CY followed by TBI was significantly better than TBI followed by CY. The survival of secondary adoptive recipients inoculated with 10(5) spleen cells obtained from mice treated with CY/TBI was 54% (57 of 105) compared with 25% (25 of 98) of recipients inoculated with 10(5) spleen cells obtained from mice treated with TBI/CY (p = 0.0001). PMID- 8401355 TI - In vitro amplification of hypervariable DNA regions for the evaluation of chimerism after allogeneic BMT. AB - The role of mixed hematopoietic chimerism in engraftment and relapse after allogeneic BMT remains unclear. To better evaluate post-transplant chimerism we used polymerase chain reaction (PCR) in vitro amplification of four single locus simple repetitive DNA sequences, all of which vary extensively in their repeat number among different individuals: variable number of tandem repeats D1S80, APOB and D17S5, and the tetranucleotide repeat F8VWF. We tested 13 cases of CML, four of multiple myeloma (MM), three of ANLL and one of B-CLL. In a sequential analysis protocol with the different loci, the donor could be distinguished from the recipient in 14 of 20 (70%) pairs with the first marker used (D1S80). When a donor of opposite sex was involved, karyotyping and Y chromosome-specific PCR were also used. With the use of the four markers, chimerism was identified in all the pairs. Mixed chimerism was present in 5 patients, and complete chimerism in 15. No patients relapsed. The application of PCR for documenting post-transplant chimerism has several advantages over Southern blotting: increased sensitivity, use of small amounts of sample, ease of preparation of DNA, elimination of restriction enzyme analysis and of radioisotopes, and speed. PMID- 8401356 TI - Epidemiology and diagnosis of invasive pulmonary aspergillosis in bone marrow transplant patients: results of a 5 year retrospective study. AB - Of 322 patients undergoing allogeneic BMT, 18 developed invasive pulmonary aspergillosis at a mean of 115 days post-transplant, with a mortality rate of 82%. Pulmonary localization was common but cerebral involvement was seen in 10 of 18 patients. The diagnosis was made ante mortem in 11 patients by direct examination of pathological samples or culture and A. fumigatus was the only species isolated. Specific antibodies were not demonstrated before or at the time of clinical symptoms and Aspergillus antigen was only seen in one patient a few days before death. PMID- 8401357 TI - Adoptive immunotherapy for recurrent CML after BMT. AB - Three patients with CML who relapsed after transplantation with T-depleted BM from their HLA-identical siblings were treated with transfusions of donor peripheral blood mononuclear cells, in combination with (short) IFN alpha 2 therapy. CML was successfully controlled as shown by the complete disappearance of Philadelphia-positive metaphases within 90 days of treatment. This treatment appears to be very effective as neither bcr-abl transcripts nor markers specific for hematological cells of recipient origin could be detected by very sensitive PCR techniques. Two patients treated in chronic phase are without evidence of disease 300 and 360 days after treatment. The third patient, treated in accelerated phase, died with BM aplasia, 39 days following PBMC infusions. Failure to detect residual donor-derived granulocytes, as was the case in this patient prior to initiating adoptive immunotherapy, may indicate loss of donor derived BM activity. This may help predict and possibly prevent the occurrence of life-threatening aplasia after successful clearance of malignant hematopoiesis. PMID- 8401358 TI - Propantheline prevention of mucositis from etoposide. AB - Mucosal toxicity is dose limiting for etoposide. This may be related to the direct effects of etoposide on the mucosa. Twelve patients receiving etoposide 1800 mg/m2 as part of a myeloablative pre-transplant regimen were randomized to receive propantheline 30 mg or placebo orally every 6 h for six doses. Mucositis was less frequent (2 of 6 vs 5 of 6) and less severe (p = 0.05) in the propantheline arm. There were no differences in tumor response or survival between the two groups. Propantheline is an anticholinergic that causes xerostomia by decreasing salivation. Propantheline may reduce the salivary excretion of etoposide and could reduce its toxic effects on the mucosa. Propantheline is effective in reducing the incidence and severity of mucositis in patients receiving high-dose etoposide. PMID- 8401359 TI - Preparation for marrow transplantation in Hodgkin's and non-Hodgkin's lymphoma using Bu/CY. AB - The results of autologous and allogeneic BMT in 44 patients with Hodgkin's or non Hodgkin's lymphomas who received preparative therapy consisting of busulfan 16 mg/kg and cyclophosphamide 120 mg/kg are presented. Sixteen patients are surviving free of disease between 8 months and 6 years after transplantation. Thirteen patients either did not attain complete remission or experienced a recurrence of their disease. Fifteen patients died from treatment-related complications. Karnofsky score < or = 70, lactate dehydrogenase greater than twice normal and the development of hepatic veno-occlusive disease were associated with failure to achieve lymphoma-free survival. PMID- 8401360 TI - High-dose cyclophosphamide-induced myocardial damage during BMT: assessment by positron emission tomography. AB - Despite its potential to cause myocardial damage, high-dose CY in doses up to 200 mg/kg is an integral part of preparative regimens for BMT. Conventional tests, such as an electrocardiogram or echocardiogram, have lacked sensitivity in prediction of cardiotoxicity in this patient population. We prospectively compared serial electrocardiograms and positron emission tomography scans before and after CY administration to investigate the possible changes in 13N-ammonia perfusion and 18F-2-deoxyglucose metabolism after CY administration in 12 consecutive patients undergoing BMT. Neither global nor regional changes in myocardial N-13 ammonia and 18-fluorodeoxyglucose were significant when compared with baseline studies and control studies (p < 0.05). In a single patient, however, a substantial increase in 13N-ammonia perfusion was seen in the inferior region simultaneously with electrocardiographic T wave inversions in the inferior leads. These changes may be due to alterations in myocardial blood flow or membrane permeability. PET scanning may be a useful adjunct in evaluating CY cardiotoxicity, although further investigations are needed to elucidate its role in clinical practice. PMID- 8401361 TI - Unrelated donor BMT for Wiskott-Aldrich syndrome. AB - The role of allogeneic sibling BMT for children with Wiskott-Aldrich syndrome is established. Mismatched T cell-depleted BMT has been successful, although significant problems with graft rejection, GVHD, and post-transplant lymphoproliferative disorders have been reported. We have performed four BMTs for children with Wiskott-Aldrich syndrome utilizing phenotypically HLA-identical unrelated donors. A non-TBI (total body irradiation) conditioning regimen was utilized, and BM was not T cell-depleted. All patients engrafted and developed significant, although manageable, GVHD. All patients are alive 3+ to 17+ months post-transplant. These results suggest that matched unrelated donor BMT has a definite role in the treatment of Wiskott-Aldrich syndrome. PMID- 8401362 TI - Fluorescence in situ hybridization with X and Y chromosome probes for cytogenetic studies on bone marrow cells after opposite sex transplantation. AB - We investigated the efficacy of fluorescence-labelled chromosome probes (CEP-X/Y) for the X and Y chromosomes to study patients who have had opposite sex BMT. These probes hybridize to the centromere region of the X chromosome and nearly the entire long arm of the Y chromosome. These probes are direct-labelled and produce X and Y signals that can be simultaneously viewed and readily distinguished from each other by color and size after only five brief washes. We investigated BM specimens from 20 normal donors and 16 patients who had undergone an opposite sex BMT. We found no significant interinvestigator differences with respect to scoring XX or XY interphase cells. The 'normal range' for XX cells in males was up to 0.628% and for XY cells in females it was up to 0.299%. Each of the specimens from the patients who underwent BMT had a significant number of donor cells compared with normal range. We suggest that an economical, rapid and accurate cytogenetic test can be achieved by using these probes as an adjunct to conventional cytogenetics. PMID- 8401363 TI - Interferon gamma is effective for BM purging in a patient with CML. AB - We report a case of Ph-positive CML where the BM was incubated for 24 h with 10(3) IU/ml IFN gamma and then cultured in liquid media for 4 weeks. After 24 h incubation, there was no differential sensitivity of CML CFU-GM to IFN gamma compared with untreated BM. Subsequent long-term culture (LTC) of the IFN gamma treated CML BM, however, demonstrated a 75% inhibition of production of CFU-GM from the second week onwards. Using PCR, we were able to demonstrate two types of BCR-ABL transcript in the diagnostic BM. After 4 weeks of LTC, the J(bcr b3/ABL II) RNA transcript persisted in the untreated BM, whereas neither BCR/ABL RNA transcripts were detected in the culture established with IFN gamma-treated CML BM. This study has two points of interest with the demonstration of (1) a possible antileukaemic effect of IFN gamma on the progenitors generated in the LTC system, and (2) the use of highly sensitive PCR technology to evaluate the effectiveness of IFN gamma to purge CML BM of Ph-positive cells. PMID- 8401364 TI - Interleukin-2 with or without lymphokine-activated killer cells as consolidative immunotherapy after autologous bone marrow transplantation for acute myelogenous leukemia. AB - IL-2 with or without autologous lymphokine-activated killer (LAK) cells, administered early after ABMT for AML may eradicate residual disease and reduce relapses. This paper reports 14 patients who received IL-2 or IL-2 plus LAK cells after ABMT for AML in first relapse or at a later stage, in two separate trials. Patients with AML in first relapse (n = 9), second CR (n = 3) or second relapse (n = 2) underwent ABMT after busulfan (BU), CY and total body irradiation (n = 11) or BU/CY alone (n = 3), with marrow that was (n = 6) or was not (n = 8) purged with 4-HC. In a previously-reported Phase I trial, eight patients received IL-2 (Roche) by continuous infusion at 0.3-3.0 x 10(6) U/m2/day x 5 days and, after 6 days of rest, 0.3 x 10(6) U/m2/day x 10 days. In a subsequent trial, five patients received IL-2 at 3.0 x 10(6) U/m2/day x 5 days, underwent leukapheresis for 3 days and received their LAK cells plus IL-2 (0.3 x 10(6) U/m2/day x 10 days). A sixth patient received only 2 days of IL-2, developed sepsis and died of multiorgan failure. All other patients had mild to moderate toxicity which was reversible. All patients developed neutrophilia, lymphocytosis and thrombocytopenia. IL-2 with or without LAK therapy was initiated 21-91 days (median 51 days) after ABMT. Severe thrombocytopenia (< 10 x 10(9)/l) occurred during the apheresis days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401365 TI - Pneumococcal arthritis after allogeneic bone marrow transplantation. AB - We report a case of pneumococcal arthritis occurring in a 15-year-old boy following allogeneic BMT. The post-transplant course was complicated by GVHD requiring prolonged immunosuppressive therapy. He experienced recurrent infections, including pneumococcal pneumonia. Thirty-five months after BMT and 12 months after the pneumococcal pneumonia, pneumococcal arthritis of the left knee occurred. This is the first reported case of arthritis caused by Streptococcus pneumoniae after allogeneic BMT. Penicillin prophylaxis may be used to prevent recurrence of pneumococcal infections in patients with chronic GVHD. PMID- 8401366 TI - HIV infection after autologous bone marrow transplantation despite HIV-antibody and HIV-antigen screening. AB - We report a 41-year-old woman who underwent ABMT for non-Hodgkin's lymphoma during her third CR. Her post-transplant course was complicated by interstitial pneumonitis, hemorrhagic cystitis, cytopenia and episodes of infection from herpes zoster virus and Staphylococcus aureus. She required prolonged blood product support and was later found to be seropositive for anti-HIV on day +191 despite HIV-antibody and HIV-antigen screening of blood donors. PMID- 8401367 TI - Cyclophosphamide cardiac injury mimicking acute myocardial infarction. AB - We report a patient in whom cyclophosphamide (CY) caused damage to the heart that was manifested by ST segment elevation and marked elevation of serum cardiac isoenzymes and mimicked myocardial infarction. Post-mortem examination did not reveal any local vascular event (e.g. thrombus or spasm) and suggested diffuse myocardial injury secondary to CY. PMID- 8401368 TI - Use of recombinant GM-CSF following allogeneic BMTs for aplastic anemia. AB - GM-CSF has been used successfully in autologous BMTs, and more recently in patients undergoing allogeneic BMT, for acute or chronic leukemia. We report two patients with hepatitis-related aplastic anemia who received recombinant human GM CSF following HLA-identical sibling allogeneic BMTs. Both patients were conditioned with CY 200 mg/kg given over 4 days and received GM-CSF at 250 micrograms/m2 beginning 6 h after marrow infusion and continuing daily until the absolute neutrophil count was > 1.0 x 10(9)/l for 2 days. Both patients had prompt engraftment, achieving an absolute neutrophil count of > 0.5 x 10(9)/l on day 13. Neither patient had side-effects attributable to the GM-CSF although one patient developed severe acute GVHD after the cessation of GM-CSF therapy. Our experience suggests that GM-CSF can be safely used in aplastic anemia patients undergoing BMT and that GM-CSF may be useful to decrease the incidence of graft failure associated with less intensive conditioning regimens. PMID- 8401369 TI - Autologous bone marrow transplantation with marrow cryopreserved for ten years. PMID- 8401370 TI - Regimen-related toxicity after BMT with an increased dose of fractionated total body irradiation. PMID- 8401371 TI - 1993 progress report from the International Bone Marrow Transplant Registry. Advisory Committee of the International Bone Marrow Transplant Registry. AB - The International Bone Marrow Transplant Registry is an organization devoted to scientific research in BMT. More than 230 transplant teams worldwide contribute detailed information about recipients of allogeneic and syngeneic BMT for study. Results of analyses are published in medical journals and presented at national and international scientific meetings (more than 60 publications and more than 500 presentations in the past 4 years). This collaborative research program has grown rapidly with more than 2000 cases reported annually. This report summarizes results of several recent investigations and reviews the state of BMT in leukemia and aplastic anemia. PMID- 8401372 TI - A new method of technetium-99m labeling of monoclonal antibodies through sugar residues. A study with TAG-72 specific CC-49 antibody. AB - We have developed a very efficient labeling technique for monoclonal antibodies with technetium-99m. Oxidation of sugar residues on the IgG class of antibodies leads to the generation of aldehyde groups which are further reacted with two newly developed hydrazide compounds. This methodology introduces sulfhydryl groups on the antibody through sugar residues which can be labeled with technetium-99m. We have studied the TAG-72 specific second generation antibody CC 49. The specific activity of the labeled antibody was high without loss of its immunoreactivity. PMID- 8401373 TI - Synthesis of 3-[18F]fluoromethyl-BTCP and evaluation as a potential PET radioligand for the dopamine transporter in baboons. AB - In an attempt to visualize in vivo the dopamine transporter and evaluate its potential as an imaging tool for monitoring dopamine fiber degeneration by positron emission tomography, the 18F-positron-emitting analogue of 3 fluoromethyl-1-[1-(2-benzothienyl)-cyclohexyl]-piperidine, [18F]BTCP, was synthesized and tested in a primate model of hemiparkinsonism. [18F]BTCP was obtained from cyclotron-produced n.c.a. [18F]fluoride (110 min half-life) and by nucleophilic substitution from 3-bromomethyl-BTCP with a radiochemical yield of 6% (decay-corrected). After intravenous injection, the cerebral distribution of the radioactivity was observed mainly in cortical areas and cerebral structures enriched in catecholamine reuptake sites such as the caudate-putamen complex and the thalamus. The binding ratio, defined with respect to the cerebellum (taken as a region of non-specific binding), was highest in the thalamus (1.42), intermediate in the putamen (1.36) and lowest in the caudate nucleus (1.17), suggesting that some specific binding occurs in these regions. After saturation of dopamine and norepinephrine transporters by nomifensine, the binding ratio in the thalamus, putamen and caudate nucleus striatum remained essentially unchanged in the non-lesioned hemisphere. When comparing binding ratios between the intact and the dopamine-denervated striatum, there was a modest loss of binding in the denervated striatum, suggesting that degeneration of dopaminergic fibers could be detected using 3-[18F]fluoromethyl-BTCP. However due to a high non-specific binding in vivo, the interest of 3-[18F]fluoromethyl-BTCP to image the dopamine reuptake system in vivo appears rather limited. PMID- 8401374 TI - Titration of the in vivo uptake of 16 alpha-[18F]fluoroestradiol by target tissues in the rat: competition by tamoxifen, and implications for quantitating estrogen receptors in vivo and the use of animal models in receptor-binding radiopharmaceutical development. AB - We have measured in vivo the uptake of 16 alpha-[18F]estradiol (FES) by target tissues in the immature rat at increasing dose levels (obtained by dilution of [18F]FES with unlabeled estradiol). This was done to examine the binding capacity of target tissues in vivo and to determine whether the uptake in receptor-rich tissues was flow limited, as this has implications concerning the appropriateness of using receptor-rich tissues in experimental animals as models for FES uptake by receptor-poor breast tumors in humans. We also wanted to establish the dose level of the anti-estrogen tamoxifen required to block target tissue uptake of FES. We found that in untreated rats, specific uptake in the uterus saturated at c. 180 pmol/g, in the ovary at c. 54 pmol/g and in the muscle at c. 2 pmol/g. At an intermediate dose of tamoxifen (570 micrograms/kg), uptake saturated at somewhat lower levels, and at a high tamoxifen dose (1710 micrograms/kg), yet lower specific uptake was evident. In the FES titrations at low dose levels of FES, both the uterus and the ovaries, but not the muscle, showed characteristics of flow-limited uptake, i.e. the uptake-to-dose ratio reached a maximum level. This flow limitation suggests that only when receptor levels are sufficiently low will the FES uptake be related to receptor concentration. While receptor-rich tissues such as the rat uterus will show this flow limitation, the receptor concentration in most primary and metastatic human breast tumors is sufficiently low, so that the uptake should parallel receptor content. In in vivo distribution studies, target tissues (or tumors) with low receptor content will be more fully saturated and ligand more readily displaced. Also, uptake by secondary target tissues (i.e. those with a lower content of estrogen receptor, such as muscle, thymus and kidney) may be better models for assessing the effectiveness of new breast tumor imaging agents than uptake by receptor-rich tissues. PMID- 8401375 TI - Metabolism of 15-(4'-[123I]iodophenyl)pentadecanoic acid ([123I]IPPA) in the rat heart; identification of new metabolites by high pressure liquid chromatography and fast atom bombardment-mass spectrometry. AB - The metabolism of 15-(4'-iodophenyl)pentadecanoic acid (IPPA) in the heart muscle is commonly believed to end at 4-iodobenzoic acid as the main and final product of beta-oxidation. However, investigation of the metabolic fate of IPPA in Langendorff rat hearts using high pressure liquid chromatography (HPLC) and negative fast atom bombardment mass spectrometry (FAB-MS) revealed new results. After perfusing isolated rat hearts with [123I]IPPA, metabolites were monitored by HPLC using simultaneous detection of gamma-radioactivity and u.v. absorbance. The identification of HPLC separated metabolites was based on their nominal molecular weights as determined by negative FAB-MS. According to these measurements five catabolites were identified with decreasing concentration: 3 (4'-iodophenyl)propanoic acid >> 3-(4'-iodophenyl)propanoic acid = 5-(4' iodophenyl)-3-hydroxypentanoic acid >> 4-iodobenzoic acid. Additionally, an anabolic metabolite was found exclusively in the lipid ester fraction. From the hydrolysed heart lipids this compound was identified as 11-(4' iodophenyl)undecanoic acid. Its formation is explained by the action of cytosolic fatty acid synthetase on IPPA catabolites. This metabolic behaviour may be of importance for the interpretation of sequential heart scintigraphy performed with [123I]IPPA. PMID- 8401376 TI - Introduction of five potentially metabolizable linking groups between 111In cyclohexyl EDTA derivatives and F(ab')2 fragments of anti-carcinoembryonic antigen antibody--I. A new reproducible synthetic method. AB - The purpose of this study was to synthesize new bifunctional linker-chelating agents for the modification of the in vivo distribution of 111In-labeled antibodies. A general simple synthetic method of preparing cyclohexyl EDTA (CDTA) derivatives containing a linker/spacer group is described. Linkers prepared included a diester, a six carbon aliphatic chain, two thioethers and a disulfide group. The CDTA-linker compounds were coupled to F(Ab')2 fragments of anti carcinoembryonic antigen monoclonal antibody and labeled with 111In with good retention of immunoreactivity. PMID- 8401377 TI - Introduction of five potentially metabolizable linking groups between 111In cyclohexyl EDTA derivatives and F(ab')2 fragments of anti-carcinoembryonic antigen antibody--II. Comparative pharmacokinetics and biodistribution in human colorectal carcinoma-bearing nude mice. AB - The five linker-containing immunoconjugates described in the preceding paper were labeled with 111In and tested for their biodistribution, pharmacokinetics and immunoscintigraphic imaging properties in tumor-xenografted nude mice. The results were compared with DTPADA and CDTAMA for reference. Results showed that, for immunoscintigraphy, the derivatives in decreasing order of effectiveness were: aliphatic (tumor/liver > 4.5 and tumor/kidney > 6.5 at 96 h), thioether (tumor/liver > 3 and tumor/kidney > 1.2 at 24 h), ethylene glycol succinate (tumor/liver > 1.7 and tumor/kidney > 0.5 at 24 h) and disulfide (tumor/liver > 0.5 and tumor/kidney > 0.6 at 96 h). Pharmacokinetic results were complementary with those of the biodistribution studies and provide a basis for the study of in vivo metabolic mechanisms of linker-immunoconjugates. Indium-111-labeled linker immunoconjugates appear promising for tumor imaging with better contrast than what is obtained with the use of the conventional 111In-DTPA dianhydride chelate. PMID- 8401378 TI - Tumor uptake of 99mTc-MIBI and 201Tl by a 9L gliosarcoma brain tumor model in rats. AB - There have been several recent case reports of the accumulation of 99mTc-MIBI [hexakismethoxyisobutylisonitriletechnetium(I), Cardiolite, Sestamibi] in tumors, but no reports of the uptake of this radiopharmaceutical in an animal model. To address this question, the biodistributions of 99mTc-MIBI and 201Tl were compared in Fisher rats bearing 9L gliosarcomas. The results showed that, although the absolute uptake of the tracers by the tumor is relatively low (< 1% ID/g), the tumor-to-normal brain ratios are greater than 6:1 because of low uptake by normal brain. The tumor-to-normal brain ratio of 99mTc-MIBI exceeds that of other currently available 99mTc radiopharmaceuticals suggesting that 99mTc-MIBI may be of particular value in the clinical evaluation of brain tumors and that further investigation of this class of compounds as tumor-avid radiopharmaceuticals is necessary. PMID- 8401379 TI - The use of [18F]4-fluorobenzyl iodide (FBI) in PET radiotracer synthesis: model alkylation studies and its application in the design of dopamine D1 and D2 receptor-based imaging agents. AB - [18F]4-Fluorobenzyl iodide ([18F]FBI) was prepared, and a series of model alkylation studies were conducted to determine its chemical reactivity toward nitrogen and sulfur nucleophiles of varying nucleophilicities. [18F]FBI was found to react rapidly with secondary amines and anilines to give the corresponding N [18F]4-fluorobenzyl analogue in high yield. Amides and thiol groups required the use of a base catalyst. The utility of [18F]FBI was documented by investigation of dopamine D1 and D2 receptor-based radiotracers. PMID- 8401380 TI - G2M arrest and apoptosis in murine T lymphoma cells following exposure to 212Bi alpha particle irradiation. AB - Asynchronous exponentially growing EL4 murine T lymphoma cells were exposed either to high LET alpha-radiation from 212Bi-DTPA or to gamma-radiation from a 137Cs source. Radiation-induced cell cycle perturbation was studied by flow cytometry. Alpha irradiation, like gamma, transiently arrested cells in the G2M phase in a dose-dependent manner. The maximum percentages of cells accumulated in G2M 18 h after alpha- and gamma-irradiation were comparable, though the dose response relationships differed. The "RBE" value for G2M block for alpha- versus gamma-radiation was approx. 4. Electron microscopic studies of the cell samples where a large proportion of cells were arrested in G2M showed subcellular changes in nuclear membrane and the presence of morphologically apoptotic cells. Biochemical analysis of DNA from irradiated cells by agarose gel electrophoresis revealed more extensive DNA fragmentation for alpha- vs gamma-irradiation, even at relatively low total doses. We conclude that the high LET radiation is more efficient in inducing G2M block and apoptosis in EL4 lymphoma cells. The overall radiosensitivity of some high and low grade malignant lymphoma cells to radiation may correlate with these processes. The clinical implications of 212Bi-induced G2M delay may be particularly important for biologically targeted high LET radiopharmaceutical therapy. PMID- 8401381 TI - Diagnosis and treatment of dissociative disorders: A multidisciplinary approach. In honor of the 100th anniversary of the birth of Karl Augustus Menninger. PMID- 8401382 TI - Dissociative processes: theoretical underpinnings of a working model for clinician and patient. AB - Dissociative symptoms are now widely recognized, but they are nevertheless challenging for clinicians and bewildering to patients. The author presents a conceptual framework for understanding dissociative processes, emphasizing discontinuity in experience and delineating various junctures in the unfolding of dissociative episodes. The conceptual framework provides the theoretical underpinnings for a simple schema that can be used to explain dissociation to patients and to help them articulate their dissociative experience. PMID- 8401383 TI - Toward a psychoanalytic understanding of multiple personality disorder. AB - The author suggests a developmental psychoanalytic frame from which to understand the clinical phenomenology of multiple personality disorder (MPD). Annihilation anxiety and fears of nonbeing are understood as central; they are seen as resulting from actual early traumatic impingements at key developmental periods. Alter "personalities" are conceptualized as functional delusional processes that serve to maintain self-cohesion. The alters are brought about through the subject's lack of capacity for illusion. Some therapeutic implications regarding a psychoanalytic stance are discussed. PMID- 8401384 TI - Selves that starve and suffocate: the continuum of eating disorders and dissociative phenomena. AB - Recent reports suggest a link between eating disorders and dissociative phenomena. The author offers a psychodynamic perspective on this connection and proposes that these disorders may occur along a continuum. Viewing these conditions as related suggests that eating disorders, like dissociative disorders, may have not only self-destructive but also self-preserving aspects. The author explores treatment implications of this perspective. PMID- 8401385 TI - Diagnosing dissociative disorders. AB - The increasing prevalence of dissociative disorders has spawned a range of diagnostic tools. The authors describe the use of several current methods- screening instruments, structured interviews, psychological testing, and hypnosis -and they advocate enlisting the patient as an active collaborator in the diagnostic process. PMID- 8401386 TI - Incorporating hypnosis into the psychotherapy of patients with multiple personality disorder. AB - Psychotherapeutic treatment of persons with multiple personality disorder frequently includes judicious use of hypnosis. The author outlines widely accepted essential features of this form of treatment: developing self-soothing techniques, "mapping" the system of alternate personalities, facilitating communication between alters and with the therapist, managing abreaction, and- when possible and appropriate--aiding the process of fusion. The author shows how dissociative processes that originally were used for sheer psychic survival can be drawn on to improve psychological health. PMID- 8401387 TI - Internal containment in the treatment of patients with dissociative disorders. AB - Both external and internal containment techniques are useful in treating patients with multiple personality disorder and other dissociative disorders. The author focuses on internal containment using various individualized strategies for helping these patients cope with life and work through past traumas. Because the visual imagery typically used in these strategies varies significantly from patient to patient, the author does not intend to provide an exhaustive description. The techniques are illustrated by case examples from the author's clinical practice. PMID- 8401388 TI - Group psychotherapy for persons with multiple personality and dissociative disorders. AB - Group psychotherapy is a valuable part of the treatment of patients with multiple personality and dissociative disorders. After reviewing the sparse literature on the subject, the author describes the use of group psychotherapy with these patients alone, as well as with two types of groups in which these patients may participate: incest survivors and general group therapy patients. She concludes by emphasizing the need for expert training for therapists who conduct group psychotherapy with multiple personality and dissociative disorder patients. PMID- 8401389 TI - Family treatment of spouses and children of patients with multiple personality disorder. AB - Although there is a large body of literature on the individual treatment of patients with multiple personality disorder (MPD), there are few accounts of the treatment of the spouses and children of these patients. After reviewing some of the existing literature, the authors present case examples of family work with patients, spouses, and children on two inpatient units and in the partial hospitalization program at Menninger. Techniques for the beginning, middle, and later phases of family treatment are illustrated. PMID- 8401390 TI - Nursing care of the self-mutilating patient. AB - Self-mutilative behavior is common among patients with multiple personality and other dissociative disorders. Nursing staff members face particular challenges in managing these patients because one act of self-mutilation can disrupt the entire inpatient milieu. The authors present an approach to nursing care that focuses on working with patients to understand and develop a specific plan to curtail the self-mutilative behavior. PMID- 8401391 TI - Multiple personality disorders: treatment coordination in a partial hospital setting. AB - Many patients with multiple personality disorder (MPD) can be effectively treated in a partial hospital setting. Partial hospital treatment teams and their patients develop and manage a safe transitional environment that contains the therapy and begins the process of independent functioning in the community. Not all patients with MPD can be treated in partial hospital settings. The author provides guidelines for appropriate placement of these patients. PMID- 8401392 TI - Transactions of the Topeka Psychoanalytic Society. PMID- 8401393 TI - Direct expression of adrenodoxin reductase in Escherichia coli and the functional characterization. AB - A plasmid for direct expression in Escherichia coli of the mature form bovine adrenodoxin reductase was constructed from the full-size cDNA for the enzyme [Y. Sagara, Y. Takata, T. Miyata, T. Hara, and T. Horiuchi, J. Biochem. (Tokyo), 102, 1333 (1987)] and an expression vector pCWori+. The recombinant adrenodoxin reductase was purified from the transformed E. coli cell lysates using adrenodoxin-Sepharose affinity chromatography [T. Sugiyama and T. Yamano, FEBS Lett., 52, 145 (1975)] with a yield of 2.5 mg/l of culture. The purified recombinant enzyme showed a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and migration was identical with that of the authentic enzyme purified from bovine adrenal cortex mitochondria. The recombinant enzyme had Ser at its amino-terminus and the sequence of the amino terminal 9 residues was identical with that of the authentic bovine enzyme. The absorption spectrum of the recombinant enzyme showed peaks at 270, 376, and 450 nm and shoulders at 425 and 475 nm. Flavin content of the recombinant enzyme was 0.8 mol FAD/mol. The apparent Km value for bovine adrenodoxin in NADPH-cytochrome c reductase activity using a reconstitution system was 16 nM, a value comparable with that of the authentic bovine enzyme (17 nM). The cholesterol side chain cleavage activity with a reconstitution system was about 75% of that obtained when the authentic enzyme was used. PMID- 8401394 TI - Alteration of fibrinogen secondary structure by cis-diamminedichloroplatinum(II) and calcium protection. AB - The structural alteration of human fibrinogen (Fbg) by cis diamminedichloroplatinum(II) (cis-DDP) was investigated using intrinsic fluorescence and circular dichroism (CD) spectra. In a preceding report, incubation of Fbg with cis-DDP under physiological conditions resulted in the cleavage of disulfide (S-S) bonds (N. Ohta, T. Yotsuyanagi and K. Ikeda, J. Pharmacobio-Dyn., 15, 611 (1992)). The intensity of fluorescence for cis-DDP treated Fbg, in which 6.7 S-S bonds per mol of protein were cleaved, was reduced to 42% relative to native Fbg. CD studies suggested that the alpha-helix content decreased from 42% in its natural state to 29% in the cis-DDP-treated form. The extent of the fluorescence quenching and the decrease in helix correlated with the number of cleaved S-S bonds. The results indicate that cis-DDP leads to a secondary conformational alteration of Fbg involving a structural perturbation of nearby tryptophan residue(s) through S-S bond cleavage. In a buffer containing calcium, however, the smaller fluorescence quenching and the smaller helix decline were observed in cis-DDP-treated-Fbg, even though the same level of S-S cleavage occurred. Calcium binding would protect Fbg in terms of structural alteration as well as S-S cleavage by both dithiothreitol (DTT) and cis-DDP. PMID- 8401395 TI - Metabolism of clentiazem in rats. AB - Following oral dosing of [14C]clentiazem to rats the metabolites in urine and bile were separated and their chemical structures were investigated by HPLC and GC-MS analyses. Fifteen basic, 6 acidic, 2 neutral and 4 conjugated metabolites were found in urine and/or bile. Eight basic metabolites (MB1-8) were identified as the synthetic compounds; deacetyl clentiazem (MB1), N-monodemethyl clentiazem (MB2), deacetyl-N-monodemethyl clentiazem (MB3), deacetyl-O-demethyl clentiazem (MB4), N-monodemethyl-O-demethyl clentiazem (MB5), deacetyl-N-monodemethyl-O demethyl clentiazem (MB6), O-demethyl clentiazem (MB7) and N-didemethyl clentiazem (MB8). The chemical structures of seven basic metabolites (MB9-15) were assigned as follows, deacetyl-N-didemethyl clentiazem (MB9), O-demethyl-N didemethyl clentiazem (MB10), deacetyl-O-demethyl-N-didemethyl clentiazem (MB11), N-monodemethyl-2-hydroxy-methoxyphenyl clentiazem (MB12), deacetyl-2-hydroxy methoxyphenyl clentiazem (MB13), deacetyl-N-monodemethyl-2-hydroxy-methoxyphenyl clentiazem (MB14) and deacetyl-N-didemethyl-2-hydroxy-methoxyphenyl clentiazem (MB15). Four acidic metabolites were identified as the synthetic compounds: (+) (2S,3S)-3-(acetyloxy)-8-chloro-3,4-dihydro-2-(4-methoxyphenyl) -4-oxo-1, 5 benzothiazepin-5(2H)-acetic acid (MA1), deacetyl-MA1 (MA2), O-demethyl-MA1 (MA3) and deacetyl-O-demethyl-MA1 (MA4); and the two remaining acidic metabolites, MA5 and MA6, were presumed to be hydroxylated MA3 and MA4, respectively. Two neutral metabolites were identified as the synthetic compounds; (+)-(2S,3S)-3-(acetyloxy) 8-chloro-3,4-dihydro-2-(4-methoxyphenyl) -4-oxo-1, 5-benzothiazepin-5(2H) acetonitrile (MN1) and deacetyl MN1 (MN2). Other two metabolites conjugated with glucuronic acid were found in bile and the structures were presumed to be 8 chloro-2,3-dihydro-3-hydroxy-5-(2-hydroxyethyl)-2-(4-hydroxyphenyl)-1, 5 benzothiazepin-4(5H)-one (MN3) and 2-methoxyphenyl MN3 (MN4). The glucuronide or sulfate of MA4 was also detected. These metabolites were formed by a number of pathways including deacetylation, deamination, N-demethylation, O-demethylation, aromatic hydroxylation and conjugation. PMID- 8401396 TI - Disposition and metabolic fate of clentiazem in rats and dogs. AB - The plasma concentrations and time courses of radioactivity and unchanged drug, the excretion of radioactivity into urine and feces, and the proportion of metabolites in plasma and urine were studied after oral administration of [14C]clentiazem to male and female rats and dogs. Apparent sex-related differences were found in the disposition and metabolism of clentiazem in rats. The plasma levels of radioactivity and acidic metabolites were higher in males than in females. The plasma levels of unchanged drug, on the other hand, were about the same in both sexes. Higher conversion of clentiazem to its acidic metabolites in the liver of male rats and higher excretion of the acidic metabolites in the urine of female rats, presumably due to sex-related differences in cytochrome P-450 and renal clearance, respectively, seem to explain these differences in the disposition of clentiazem in male and female rats. No suggestion of a similar sex difference was found in dogs. The plasma concentrations and time courses of radioactivity and unchanged drug in male dogs were similar to those in female dogs, and the excretion of radioactivity in both sexes was also similar. The main plasma metabolite in male and female dogs was O demethyl clentiazem (MB7). A species difference between rat and dog was suggested, since the major metabolic pathways were different and no sex difference was found in dogs. PMID- 8401397 TI - Effect of beta-alanyl-L-histidinato zinc on bone metabolism in rats with adjuvant arthritis. AB - The effect of a new zinc compound, beta-alanyl-L-histidinato zinc (AHZ), on osteopenia was investigated in rats with adjuvant arthritis. Arthritis was induced in female rats by administering 1% Mycobacterium butyricum (MB) into the subplantar surface of the right hind paw. AHZ (10, 30 and 100 mg/kg body weight) was orally administered to MB-treated rats 28 times at 24-h intervals, and the rats were bled 24 h after the last administration. Treatment with MB caused a remarkable increase in paw volume and a corresponding decrease in the ratio of albumin per globulin in serum, indicating that the treatment induces inflammation. These alterations were not significantly changed by the administration of AHZ (10, 30 and 100 mg/kg). Serum calcium and zinc concentrations are significantly decreased in rats with adjuvant arthritis. These decreases were completely restored by the administration of AHZ (30 and 100 mg/kg). Furthermore, the inflammation-induced decreases in alkaline phosphatase activity and calcium content in the femoral diaphysis were clearly blocked by the administration of AHZ (30 and 100 mg/kg). Also, the larger doses of AHZ (30 and 100 mg/kg) produced a significant increase in femoral-diaphyseal deoxyribonucleic acid and in the zinc content in rats with adjuvant arthritis. These results suggest that AHZ has a stimulating effect on bone formation in the femoral diaphysis of rats with adjuvant arthritis, although the compound did not have an anti-arthritic effect. PMID- 8401398 TI - Adriamycin-induced lipid peroxidation of erythrocyte membranes in the presence of ferritin and the inhibitory effect of ceruloplasmin. AB - When erythrocyte membranes were incubated with adriamycin (ADM) in the presence of ferritin, lipid peroxidation occurred with release of iron from the ferritin. In the presence of apoferritin, ADM did not cause lipid peroxidation. Deferoxamine inhibited the ADM-induced lipid peroxidation in the presence of ferritin. These results indicate that lipid peroxidation depends upon the release of iron from ferritin. Even when the iron content in ferritin was very low, ADM could induce lipid peroxidation. Superoxide dismutase, catalase and hydroxyl radical scavengers did not substantially affect lipid peroxidation, indicating that the peroxidation reaction was independent of superoxide, H2O2 and hydroxyl radicals. Ceruloplasmin, a ferroxidase, markedly inhibited lipid peroxidation but did not affect the release of iron from ferritin. ADM-Fe(3+)-binding erythrocyte membranes were readily formed during the incubation of erythrocyte membranes with ADM in the presence of ferritin, and deferoxamine removed iron from the ADM Fe(3+)-binding membranes, indicating that the iron moiety of the ADM-Fe(3+) complex is exposed at the membrane surface. These results may suggest that the peroxidation reaction occurs in a site-specific manner. PMID- 8401399 TI - Pharmacological activity and structure-activity relationship of (+/-)-erythro mefloquine and related compounds on the isolated mouse phrenic nerve diaphragm preparation. AB - The effects of the antimalarial agent, (+/-)-erythro-mefloquine and related compounds [(+/-)-threo-mefloquine, (+/-)-erythro-N-methylmefloquine and its N oxide, quinine, WR 184806 and halofanthrine] on the isolated mouse phrenic nerve diaphragm preparation were investigated. Based on their pharmacological effects, these compounds may be divided into two groups. The group I compounds, comprising (+/)-erythro-mefloquine, (+/-)-threo-mefloquine and WR 184806, were found to exert a contractile effect on the muscle and also to inhibit the indirectly (nerve) stimulated and directly (muscle) stimulated (after alpha-bungarotoxin) twitch responses. The group II compounds, comprising the other compounds except halofanthrine, lacked a contractile effect on muscle but potentiated the directly stimulated twitch responses (after alpha-bungarotoxin). Halofanthrine did not elicit any response from the preparation. The minimum energy conformations of these compounds were determined using an interactive molecular modelling program which incorporates MMX force field for molecular mechanics calculations. Conformational analyses of the erythro and threo isomers of mefloquine hydrochloride were also undertaken using 1H-NMR. The 1H-NMR data supported the proposal made on the basis of MMX calculations that the erythro isomer exists in solution as one predominant conformer whereas the threo isomer is present in solution as a mixed population of two stable conformers. The structure-activity relationship of the compounds is discussed. PMID- 8401400 TI - Pharmacological and pharmaceutical properties of freeze-dried formulations of egg albumin, indomethacin, olive oil, or fatty acids. AB - Formulations consisting of egg albumin, indomethacin (IND), and olive oil or fatty acids, were prepared by vigorous stirring using a high-speed homogenizer and subsequent freeze-drying. To confirm the anti-inflammatory properties and ulcerogenic effects of the formulations, we examined the action of the formulations on carrageenan-induced edema in rats as well as their ulcerogenic actions in the same species. Compared with IND alone, albumin-IND-olive oil (9:1:4.3), albumin-IND-linolenic acid (9:1:4.3), albumin-IND-linolic acid (9:1:4.3), albumin-IND-oleic acid (9:1:4.3), albumin-IND-stearic acid (9:1:4.3), and albumin-IND-tristearin (9:1:4.3) formulations all exhibited a more potent inhibitory effect on carrageenan-induced edema. In addition, the inhibitory effects on edema formation of an albumin-IND (9:1) complex was as strong as that of IND alone. These results suggested that the bioavailability of IND was increased by olive oil, fatty acid, and tristearin as absorbefacient agents. The increase in the bioavailability was evident from the fact that the mean plasma levels, maximum plasma levels (Cmax), and area under plasma concentration-time curve (AUC) values after oral administration of the albumin-IND-olive oil (9:1:4.3) formulation was significantly greater than that after administration of the drug alone. With respect to their ulcerogenic properties, the formulations were significantly less active than IND alone, suggesting that a reduction in the ulcerogenic activity of IND was by produced complexation with egg albumin. PMID- 8401401 TI - Pharmacological and pharmaceutical properties of freeze-dried formulations of egg albumin, indomethacin, olive oil, or fatty acids. II. AB - To confirm the increased bioavailability of indomethacin (IND) when incorporated in a preparation with egg albumin and olive oil, we studied the detailed pharmaceutical characteristics of a ternary formulation consisting of egg albumin, IND and olive oil. From the results of X-ray powder diffraction measurements, the drug in the formulation was found to be in an amorphous form. When orally administered to rats, the ternary formulation significantly increased the plasma concentration and cumulative biliary and urinary excretion of IND alone as well as the urinary excretion of its major metabolite, desmethylindomethacin, compared with the drug alone. In addition, the dissolution rate of IND from the formulation was higher than that of the drug alone. These results clearly suggest that the bioavailability of IND was markedly improved by incorporating it in a protein-drug formulation containing olive oil as an absorbefacient element, and this effect may be due to an increased absorption of IND. PMID- 8401402 TI - Comparative study on the accelerative effect of "koushikon" and "nanshikon" and their constituents on proliferation of granuloma tissue in rats. AB - This study was carried out to compare the accelerative effect in ether extracts of "Koushikon" and "Nanshikon" on proliferation of granuloma tissue in rats, and to elucidate this effect on optical isomer of naphthoquinone derivatives in those extracts. The content of total naphthoquinone derivatives in the ether extracts of Koushikon and Nanshikon were found to be 56.1% and 25.4%. Among naphthoquinone derivatives, Koushikon contained mostly acetyl derivative and Nanshikon mostly teracryl derivative. The percentage of R-type (shikonin-type) in total naphthoquinone derivatives of the extracts was 85.5% and 3.8%. Each ether extract showed a dose-dependent acceleration on the cotton pellet-induced granuloma formation. Comparison with corresponding doses containing the same quantity of naphthoquinone derivatives showed the accelerative potency of ether extracts of Koushikon and Nanshikon to be about the same. The result suggests that the accelerative effect on proliferation of granuloma tissue depends primarily on the total content of naphthoquinone derivatives, and not on the ratio of the optically active isomers. PMID- 8401403 TI - Diagnosis of drug allergy by the lymphocyte stimulation test with the MTT [3-(4,5 dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. AB - The lymphocyte stimulation test (LST) is one of the most useful laboratory tests for the identification of allergy to a specific drug. The present study was conducted to examine utility of the LST using the [3-(4,5-dimethyl thiazol-2-yl) 2,5-diphenyl tetrazolium bromide] (MTT) assay as a method for diagnosing drug allergy. The basic experimental conditions for the MTT assay were determined by using CTLL cells from human T-lymphoma, dependent on IL-2, and human peripheral blood lymphocytes. The amount of MTT needed was 0.5 mg per well and the peak maximum of MTT formazan was close to 565 nm. The blastogenesis of the lymphocytes was expressed as a Stimulation Index (SI). The value of the SI was almost constant at cell numbers ranging from 5 x 10(5) to 1 x 10(6) cells/ml. The SI of some drugs using normal peripheral blood lymphocytes was less than 1.3. We used the MTT assay in the LST to examine the allergenicity of 83 drugs which were used in 43 cases of suspected drug-induced hepatitis. The range of SI was 0.92-2.02. An SI of 1.2 or more was seen in 62.8% of cases, 1.3 or more in 51.2% of cases, and 1.4 or more in 41.9% of cases. There were 26 drugs with SI greater than 1.3. Seven of these were antibiotics, while the rest included antihypertensives, analgesics, psychotropics, antiallergics and antiepileptics. This method is extremely well suited for the diagnosis of drug allergy in a clinical situation. The greatest benefit of the MTT assay is that the complete test procedures can be carried out in a general laboratory. PMID- 8401404 TI - Analysis of skin penetration enhancement based on a two-layer skin diffusion model with polar and nonpolar routes in the stratum corneum: dose-dependent effect of 1-geranylazacycloheptan-2-one on drugs with different lipophilicities. AB - The effects of 1-geranylazacycloheptan-2-one (GACH) on the in vitro skin penetration of seven drugs with various lipophilicities were studied. The penetration of drugs from aqueous vehicle through the guinea pig skin was increased depending on the pretreatment dose of GACH. The largest enhancement was observed for 5-fluorouracil (5-FU) and a bell-shaped relationship was obtained between enhancement ratio and the octanol/water partition coefficients of drugs. Further, Laplace transformed equations for percutaneous absorption of drug were derived from Fick's second law based on a two-layer skin diffusion model with polar and nonpolar routes in the stratum corneum. Through curve-fitting of their equations to penetration profiles using a nonlinear regression program MULTI(FILT) combined with a fast inverse Laplace transform (FILT) algorithm, the action mechanism of GACH was discussed in terms of its effect on the partitioning and diffusivity of drugs in each domain. With an increase in pretreatment dose of GACH, the estimated partition parameters of drugs into the nonpolar route increased but their diffusivities were little affected. The analysis based on a linear free-energy relationship suggested that the increase in partitioning of drugs into the nonpolar route was explained by its increasing polarity with GACH pretreatment. PMID- 8401405 TI - Studies on zoospore attracting activity. II. Synthesis of isoflavones and their attracting activity to Aphanomyces euteiches zoospore. AB - Nineteen isoflavones were synthesized and their attracting activity to Aphanomyces euteiches zoospore were investigated. Isoflavones (1, 2, 4, 16 and 19) with an unsubstituted B ring were synthesized by the oxidation of corresponding 2'-hydroxychalcones with thallium (III) nitrate trihydrate in methanolic perchloric acid. A hydroxyl group at the C-5 position in isoflavones was necessary for strong attraction to A. euteiches zoospore. The introduction of an additional hydroxyl group at the C-7 or C-4' position strengthened the attracting activity. Moreover, the methylation of the C-7 hydroxyl group strengthened the attracting activity, but the methylation of the C-4' hydroxyl group slightly weakened it. The naturally occurring isoflavones (9 and 10), which were reported to possess estrogen activity, showed moderate attracting activity. PMID- 8401406 TI - Erythrocyte uptake and protein binding of cyclosporin A (CyA) in human blood: factors affecting CyA concentration in erythrocytes. AB - To further the understanding of the complexity of cyclosporin A (CyA) pharmacokinetics, we conducted an erythrocyte uptake and efflux study, and a protein binding study in human blood. The uptake study showed that the transport of CyA from the extracellular fraction to erythrocytes was retarded by increased human serum albumin (HSA) and lipid levels in this fraction. In addition, the concentration of CyA in erythrocytes increased with increases in CyA concentration in blood and reductions in hematocrit. The efflux study showed that the transport of CyA from erythrocytes to the extracellular fraction was essentially enhanced by increases of HSA and lipid levels in that fraction, but that these effects were relatively small. There were two affinity binding sites for CyA in ghost-free erythrocyte hemolysate, but not in the plasma fraction. The affinity binding constants for these binding sites were reduced by elevations in temperature, and under physiological conditions, 37 degrees C, almost all the CyA in erythrocytes was bound to a CyA binding protein, namely, cyclophillin. These findings suggest that CyA distribution in blood is of two different types which are present in the erythrocyte and plasma fractions, respectively. Monitoring of blood biochemistry variables showed that the concentration of CyA in erythrocytes had an interlocking relationship with these physiological factors, which were related to patient disease state, i.e., hematocrit, lipids, albumin, and total protein; the concentration of CyA in erythrocytes could be predicted from these physiological factors. PMID- 8401407 TI - Inhibition of DNA topoisomerase I activity by diethylstilbestrol and its analogues. AB - Diethylstilbestrol and its derivatives were tested to determine their inhibitory effects on the relaxation activity of DNA topoisomerase I using cell lysates from Chinese hamster V79 cells. Among the derivatives investigated, (+/-)-indenestrol B, (+/-)-IB, showed the strongest inhibitory effect. PMID- 8401408 TI - Characteristics and localization of a determinant conferring partial macrolide resistance in Staphylococcus aureus carrying plasmid. AB - Staphylococcus aureus TPR-27, a clinically isolated strain, showed constitutive resistance to some macrolide antibiotics (erythromycin, oleandomycin, spiramycin, and josamycin), but susceptibility to the other macrolide (tylosin, rokitamycin, and mycinamicin), lincosamide, and streptogramin type B antibiotics (PM resistance). The PM-resistant strain TPR-27 has carried for visible plasmids. Attempts to eliminate the resistant determinant in terms of ethidium bromide (about 3 micrograms/ml) did not succeed, and every trial to transduce the PM resistant determinant into rec- mutant ISP105 using phage 80L2 propagated on strain TPR-27 also failed at the frequency of less than 1.1 x 10(-10) transductants per plaque-forming unit. These results suggest that the PM resistance determinant is localized in chromosomal DNA. PMID- 8401409 TI - Modulation by endothelin-1 of tissue plasminogen activator and plasminogen activator inhibitor-1 release from cultured human vascular endothelial cells: interaction of endothelin-1 with cytokines. AB - The interaction of endothelin-1 (ET-1) with either interleukin-1 beta (IL-1 beta) or tumor necrosis factor alpha (TNF alpha) on the release of tissue plasminogen activator antigen (t-PA:Ag) and plasminogen activator inhibitor-1 antigen (PAI 1:Ag) was investigated in a culture system of vascular endothelial cells derived from human umbilical vein. The t-PA:Ag release was significantly decreased by either IL-1 beta or TNF alpha; ET-1 intensified the suppressive effect of the cytokines. In contrast, PAI-1:Ag release was significantly increased by either IL 1 beta or TNF alpha; ET-1 significantly reduced the stimulatory effect of the cytokines. The data suggest that endothelial cell-mediated fibrinolysis may be modulated by ET-1. PMID- 8401410 TI - Inhibitory effects of tannins on NADH dehydrogenases of various organisms. AB - We examined the effects of purified tannins and related compounds (33 species) on NADH-ubiquinone-1 oxidoreductase activity in four kinds of organism (Paracoccus denitrificans, Bacillus subtilis, Photobacterium phosphoreum, and Thermus thermophilus HB-8) and rat liver mitochondria. In addition to pentagalloylglucose, which was reported as a potent inhibitor of NADH dehydrogenases (NDH), sanguiin H-11, oolonghomobisflavan A, and polymerized procyanidin are potent inhibitors for both types of NDH (NDH-1 and NDH-2). We found that some other tannins contained in tea are also inhibitors of NDH from all organisms. PMID- 8401411 TI - Effect of divalent metal ions on the binding of thyroxine to bovine serum albumin as measured by fluorescence. AB - The binding of thyroxine (T4) to bovine serum albumin (BSA) has been studied in the presence and absence of Ca2+, Cu2+ and Zn2+ ions at various pH's in 0.1 M Tris-acetate buffer at 25 degrees C using the fluorescence method. In the presence of 50 microM Ca2+ and Zn2+ and the absence of metal ions, the binding constant (K) increased similarly with increasing pH values from pH 5 to pH 9, and the K value near midpoint, pH 7.4, was 1.66 +/- 0.17 x 10(6) M-1. By contrast, the binding constant remained constant between pH5 and pH9 in the presence of 10 microM Cu2+, with an average value of 1.61 +/- 0.22 x 10(6) M-1, suggesting a significant influence of Cu2+ ions on T4 binding to BSA. PMID- 8401412 TI - Pharmacokinetic interaction of zonisamide in rats. Effect of zonisamide on other antiepileptics. AB - The effects of zonisamide (ZNS) on the pharmacokinetics of phenobarbital (PB), valproic acid (VPA), carbamazepine (CBZ) and phenytoin (PHT) were investigated in rats. Additionally, the influence of ZNS on the serum protein binding, erythrocyte distribution and metabolism of these antiepileptics was studied in vitro. The t 1/2 and AUC values of PB were significantly increased by ZNS coadministration, and a significant decrease in the Vd/F value of PHT was observed after multiple dosing of ZNS. By contrast, ZNS showed no significant effect on VPA and CBZ kinetics. No significant effect of ZNS was observed in the serum protein binding, erythrocyte distribution or metabolism of other antiepileptics. These results suggest that ZNS has little effect on the pharmacokinetic behaviors of other antiepileptic drugs. PMID- 8401413 TI - Biosynthesis in vitro of 2-(3-amino-3-carboxypropyl)-isoxazolin-5-one, the neurotoxic amino acid in Lathyrus odoratus. AB - 2-(3-Amino-3-carboxypropyl)-isoxazolin-5-one (ACI), a neurotoxic amino acid from Lathyrus odoratus, was confirmed to be derived enzymatically from S-adenosyl-L methionine (SAM) and isoxazolin-5-one. Some properties of an enzyme in the biosynthesis of ACI are described. PMID- 8401414 TI - Inactivation of renin substrate by soybean trypsin inhibitors: implications for measurement of circulating inactive renin. AB - Semipurified soybean trypsin inhibitor added to rat and human plasma leads to a concentration dependent decrease in the rate of angiotensin I generation. This inhibition is due to binding of renin substrate to the inhibitor. Renin substrate present in nephrectomized rat plasma was more susceptible to binding than substrate of the normal rat suggesting structural differences in the substrate generated following nephrectomy. Because trypsin inhibition is necessary for measurement of active and inactive renin, we examined several alternate trypsin inhibitors. The Bowman-Birk inhibitor from soybean had similar actions as purified soybean trypsin inhibitor while trypsin inhibitors from lima bean and chicken did not depress renin substrate, but did have variable effects on the measured levels of active and total plasma renin. Surprisingly, crude soybean trypsin inhibitor did not suppress renin substrate and actually increased angiotensin I generation during PRA and PRC measurements. Since the crude preparation did not suppress renin substrate, changes in the specificity of the inhibitor may occur during its purification. The augmentation of PRA and PRC may be related to angiotensinase inhibitory actions. PMID- 8401415 TI - Platelet activity and salt sensitivity in the pathogenesis of systemic (essential) hypertension in black Africans. AB - Black essential hypertensive patients with a mean arterial pressure of 125 +/- 3 mm Hg (mean +/- SEM), and age and sex matched normotensive subjects with a mean arterial pressure of 89 +/- 2 mm Hg were studied under baseline conditions, after five days of salt restriction and after five days of salt loading. Salt sensitivity was defined as an increase of mean blood pressure exceeding 5% when progressing from low to high sodium intake. In vitro platelet responsiveness was assessed by aggregometry, and in vitro platelet activity by estimation of beta thromboglobulin (BTG) in plasma and thromboxane B2 (TXB2) excretion rate. Salt sensitivity was present in 66% of hypertensive and 55% of the normotensive subjects. An increased platelet aggregability to ADP (25%), to epinephrine (34%) and to collagen (12%) was found in parallel with an increased in vivo platelet activity (BTG increased by 55% and TXB2 by 18%) in the hypertensives. All changes were significantly exaggerated in the salt sensitive as compared to salt resistant hypertensive patients. PMID- 8401416 TI - The Prague Hypertensive Rat: a new model of genetic hypertension. AB - Several animal models of genetic hypertension have been developed but not all of them possess a closely related control strain. Therefore, a new model based on Wistar rats is described in which both hypertensive and normotensive lines were bred from a single parental pair. Several basic data on the two lines (called the Prague Hypertensive Rat, PHR, and the Prague Normotensive Rat, PNR) are given. PNR had a longer survival compared with PHR. At the age of 7 weeks, systolic blood pressure was 161 +/- 14 mmHg in PHR males and 109 +/- 9 mmHg in PNR males. Its further increase with age was very slow in PNR but very steep in PHR. Typical left ventricular cardiac hypertrophy developed in PHR in which cardiac output was not significantly different from that of PNR but total peripheral resistance was higher. Kidney weight was also greater in PHR than in PNR. There was no difference in basic renal functions except of proteinuria which was higher in PHR than in PNR. No differences were observed in extracellular and interstitial fluid volumes whereas plasma and blood volumes were slightly but significantly greater in PHR than in PNR suggesting a shift of extracellular fluid towards the intravascular compartment. This hypertensive model the parameters of which resemble to those of human essential hypertension should be especially suitable for cross-transplantation studies. PMID- 8401417 TI - Plasma triglycerides and 24 hour urinary sodium excretion in elderly hypertensives. A pathogenetic connection? AB - Hypertension among the elderly generally represents a salt- sensitive state. However this salt-sensitivity does not appear to result from age-related increase in either sodium or salt intake. Since 20 years new trends seem to relate the role of sodium in the genesis of hypertension to a primary abnormality of electrolyte transport of cell membrane. Lipid abnormalities have also been described in untreated patients with high blood pressure. Plasma triglycerides were considerably higher (p < 0.01) in the hypertensives than in the controls. 24 hour sodium excretion was significantly lower (p < 0.0001) in hypertensives than in the controls. We have found a strong correlation among reduced sodium excretion, higher triglycerides and elevated blood pressure in the elderly. The blood pressure correlated negatively with 24 hour sodium excretion (p < 0.0001 for systolic and p < 0.002 for diastolic) and positively with plasma triglycerides (p < 0.0001 for systolic and p < 0.001 for diastolic). The poor literature regarding an association of these two alterations in human hypertensives makes our results provocative. We speculated that these alterations may be a facet of the insulin resistance commonly detectable in human hypertensives. However, further investigations are required to answer to this intriguing hypothesis. PMID- 8401418 TI - Changes in extracellular Ca2+ over a physiologic concentration range differentially modulate reactivity of resistance arteries of spontaneously hypertensive and normotensive rats. AB - Norepinephrine-induced contractile responses of mesenteric resistance arteries of normotensive Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats were examined in the presence of 1.25, 1.5, and 2.5 mM extracellular Ca2+. In endothelium-intact WKY segments, the three levels of extracellular Ca2+ had no effect on sensitivity to norepinephrine. However, the response of SHR vessels to low dose norepinephrine was significantly enhanced in the presence of 1.25 and 2.5 mM Ca2+ versus 1.5 mM Ca2+. After endothelial WKY vessels responded like SHR vessels, i.e. sensitivity to norepinephrine was enhanced in 1.25 mM Ca2+ compared with 1.5 mM Ca2+. This response to alterations in extracellular Ca2+ was also observed after blockade of the neuronal amine pump with cocaine. These results indicate that vascular smooth muscle of both SHR and WKY respond to small physiologic changes in extracellular Ca2+ with altered sensitivity to norepinephrine. This effect is independent of the neuronal amine pump. We conclude that the endothelium of the normotensive rat protects against Ca(2+) induced changes in sensitivity to norepinephrine and this protection is absent in SHR. PMID- 8401419 TI - Neuroanatomical differentiation in the brain of the spontaneously hypertensive rat (SHR). I. Volumetric comparisons with WKY control. AB - A series of measurements was made to assess the morphology of the brain of the spontaneously hypertensive rat (SHR). The SHR brain was smaller than that of age matched normotensive Wistar-Kyoto (WKY) controls in a majority of measures of external surface landmarks. This reduction in size was evident in the youngest age group examined (94 days) and persisted in older groups (170, 240 and 350 days). The brain of the SHR was also smaller in terms of brain weight and brain weight:body weight ratios. Section-by-section digitized analyses of coronal histologic sections from 94-day-old rats revealed significant reductions in mean cross-sectional area and volume of midbrain/pons (10%) and hindbrain (11%) regions, but not of forebrain, in the SHR. Alterations in the mediolateral dimension, particularly within the pontomedullary brainstem, accounted for more of these volumetric changes than those in the dorsoventral dimension. Using the same coronal sections, it was found that surface areas and volumes of five individual nuclei/fiber tracts, selected for their involvement in central cardiovascular regulation, were significantly decreased in the SHR. The largest reduction in volume (30%) was found in the nucleus tractus solitarius, the primary site of termination of afferent baroreceptor fibers. No differences in surface area or volume were found in that portion of the cerebroventricular system (aqueduct of Sylvius) associated with the periventricular grey region, or in the inferior colliculus, which is not thought to be involved in cardiovascular control. These observations not only have practical implications, but suggest that the pathophysiological condition expressed as spontaneous hypertension in this widely-used model may be related to morphological alterations in the central nervous system. PMID- 8401420 TI - Comparison of repeated measurement of ambulatory and clinic blood pressure readings in isolated systolic hypertension. AB - Repeated clinic blood pressure measurement was compared with non-invasive ambulatory blood pressure monitoring in 10 elderly subjects with persistent isolated systolic hypertension and 11 normotensive controls. Mean clinic blood pressures +/- Standard Deviation (SD), at visit 1 were 165/81 +/- 12/7 mmHg and 136/87 +/- 14/10 mmHg respectively. Subjects were assessed on three subsequent occasions at weekly intervals. None were receiving antihypertensive or vasoactive medication. Clinical systolic and diastolic blood pressures were consistently higher than the corresponding mean daytime ambulatory blood pressures in both clinical groups at each of the visits. The difference was greater between the systolic pressures than between the diastolic pressures and these differences in systolic pressures were greater in the isolated systolic hypertensives (26 +/- 5 mmHg) than in the normotensives (7 +/- 18 mmHg). Daytime ambulatory readings aggregated from all four visits were normally distributed for both blood pressure components in both clinical groups. In the isolated systolic hypertension group the clinic systolic and diastolic blood pressure measurements corresponded to the 93rd and 85th percentiles of the ambulatory pressures respectively whereas in the normotensives the equivalent percentiles were 69 and 78. These results suggest a pressor response may largely account for the elevated systolic blood pressure seen in elderly subjects with sustained isolated systolic hypertension based on clinic readings. PMID- 8401421 TI - An experimental anti-idiotype vaccine mimicking lipopolysaccharide gives protection against Pasteurella multocida type A infection in mice. AB - An anti-idiotype strategy was employed which showed that polyclonal anti-idiotype antibodies could be produced which could mimic a linear Pasteurella multocida lipopolysaccharide (LPS) molecule. These antibodies when used as vaccine antigens, induced antibodies which recognised LPS and imparted acquired protection upon syngeneic vaccinates challenged with homologous organisms. PMID- 8401422 TI - Infection of differentiated U937 cells by Salmonella typhimurium: absence of correlation between oxidative burst and antimicrobial defence. AB - The human histiocytic lymphoma cell line U937 can be induced to differentiate along the monocyte/macrophage pathway by either phorbol myristate acetate (PMA) or by the combination of retinoic acid (RA) and 1,25-dihydroxyvitamin D3 (VD). U937 cells treated with either PMA or RA/VD were able to phagocytose Salmonella typhimurium in the presence of non-immune human serum. However, only cells differentiated by RA/VD were capable of developing an oxidative metabolic burst in response to infection. Since the oxidative burst is considered to be a potent antimicrobial mechanism, we investigated its effect on S. typhimurium. The oxidative burst failed to affect either the viability or the multiplication of S. typhimurium suggesting that it plays only a minor role in the host defence against S. typhimurium. PMID- 8401423 TI - Structural features involved in the mitogenic activity of Bordetella pertussis lipopolysaccharides for spleen cells of C3H/HeJ mice. AB - Spleen cells from the C3H/HeJ mouse strain cannot be stimulated by many smooth type lipopolysaccharides (LPSs), and by the main biologically-active region (lipid A) of these molecules. The genetic origin of this defect (expression of the mutant allele Lpsd at the chromosome 4 locus) was established over 20 years ago, but its biochemical nature has remained undefined. Several investigators have noted, however, that some particular LPSs can bypass this defect, and stimulate the proliferation of C3H/HeJ B lymphocytes. In this study we compare the mitogenic activities of the LPSs isolated from a wild strain (1414) and from a mutant 'rough' strain (A100) of Bordetella pertussis. Both LPS-1414 and LPS A100 were mitogenic for C3H/HeJ spleen cells, but their lipid A fragments were not. This indicates that a carbohydrate structure proximal to lipid A is involved in the mitogenic activity. However, the isolated polysaccharides were not mitogenic. Four sugars are common to both LPS-1414 and LPS-A100: an heptose, and three sugars bearing free amino groups. After removal of these four sugars from the LPSs by nitrous acid treatment, the recovered lipooligosaccharides were not mitogenic in Lpsd spleen cells. The results suggest that substructures present in lipid A and in this group of four sugars are both required for induction of a mitogenic effect in Lpsd splenocytes, whereas lipid A alone can stimulate Lpsn spleen cells. PMID- 8401424 TI - Serological response in rabbits to Listeria monocytogenes after oral or intragastric inoculation. AB - The serological response in rabbits against Listeria monocytogenes after oral or intragastric inoculation was investigated. Both the number of sero-positive animals and the average serum titres were higher in animals inoculated by the oral route. This difference was especially marked in rabbits inoculated with the lower dose (1 x 10(3) colony-forming units (cfu)), which developed a strong serological response (average serum titre of 1280 after 4 inoculations) in most of the inoculated animals (80%), without any clinical signs. The implication of these results in the epidemiology of listeriosis is discussed. PMID- 8401425 TI - Multiple binding of type 3 streptococcal M protein to human fibrinogen, albumin and fibronectin. AB - M proteins are major virulence factors of group A streptococci which enable the bacteria to resist phagocytic attack. Their binding capacity for different plasma proteins seems to be one reason for the antiphagocytic activity of M protein. In the present study we demonstrate that M3 protein, isolated from the streptococcal culture supernatant of strain 4/55, and the recombinant form (rM3), purified from an E. coli lysate after cloning in phage lambda-EMBL3, show a multiple binding to fibrinogen, albumin and fibronectin in Western blot and dot binding assays. Binding of M3 protein to the multifunctional extracellular matrix and plasma protein fibronectin may not only influence phagocytosis but may also contribute to the adherence of these bacteria to endothelial and epithelial cells. PMID- 8401426 TI - Interferon-gamma, interleukin-1 and tumour necrosis factor-alpha synthesis during experimental murine staphylococcal infection. AB - Several exotoxins of Staphylococcus aureus were shown to modulate the host immune system by stimulation of monokine release. BALB/c mice infected intravenously (i.v.) with live cells if S. aureus, strain Cowan 1, had a detectable serum level of TNF-alpha at 3, 4 and 5 h after injection. When S. epidermidis (strain E3380, clinical isolate) was used to infect mice, the level of TNF-alpha was lower (the detection limit of the cytotoxicity assay with WEHI cells was 40 pg ml-1). Kinetics of TNF synthesis was different from that observed in experimental infections caused by Gram-negative bacteria. Similarly to TNF-alpha, IL-1 alpha appears in a measurable level at 3 h after i.v. injection of bacteria. The highest serum level of IFN-gamma was observed 12 h after infection with both S. aureus and S. epidermidis. A quantity ten times more of S. epidermidis than of S. aureus cells was required to induce similar levels of TNF-alpha and IFN-gamma. Recombinant IFN-gamma administered in vivo in four daily doses followed by infection of S. aureus resulted in increased elimination of bacteria from the spleen, liver and peritoneal cavity of mice. PMID- 8401427 TI - Dextran sulphate enhancement of lipopolysaccharide-induced tumour necrosis factor alpha production by murine peritoneal macrophages: correlation with macrophage blockade. AB - Proteose peptone-induced murine peritoneal macrophages (M phi) were preincubated with 100-800 micrograms/ml of dextran sulphate (DS) 500 (M(r) 500,000) or DS1000 (M(r) 1,000,000). After 2-24 h of the preincubation, the M phi were stimulated with 1 microgram/ml of lipopolysaccharide (LPS) in vitro for 18 h in DS-free culture medium. The culture supernatants were then collected for TNF assay. The LPS-induced TNF activity of M phi supernatant preincubated with DS500 or DS1000 for 6 h was enhanced by up to about ten-fold compared with those preincubated without DS. This enhancing effect was not observed when M phi were preincubated with 100-800 micrograms/ml of low molecular weight DS5 (M(r) 5,000) or neutral dextran (Dex) 500 (M(r) 500,000). The enhancement of LPS-induced TNF-alpha production from M phi was observed after 2 or 4 h of incubation with DS1000 or DS500, respectively. The phagocytic activity of M phi was determined in vitro by the ingestion index and phagocytic capacity using Saccharomyces cerevisiae. Treatment with DS500 or DS1000 significantly suppressed the phagocytic activity from 2 h after the incubation, but this suppression was not observed in M phi incubated with DS5 or Dex500. Our experiments indicate that DS500 and DS1000 act directly on M phi and enhance LPS-induced TNF-alpha production from M phi, and that the enhancement is closely related to the suppression of M phi phagocytic function. PMID- 8401428 TI - Differential persistence, immunogenicity and protective capacity of temperature sensitive mutants of Salmonella enteritidis after oral or intragastric administration to mice. AB - The persistence of Salmonella enteritidis temperature-sensitive (ts) mutants of different phenotypes in Peyer's patches (PP) and the spleen, and their immunogenicity after intragastric (i.g.) and peroral (p.o.) administration to mice was investigated. After p.o. administration the ts mutant C/2/2 colonized PP, but was not recovered from the spleen. After i.g. administration the ts mutant E/1/3 colonized both the spleen and PP for at least 2 weeks. Mutant C/2/2 persisted in PP up to 8 days but was not found in the spleen. Mutant H/2/26, although it poorly colonized the PP, was recovered from the spleen up to day 15 after i.g. administration. Immunization with E/1/3 by either the i.g. or the p.o. routes protected mice from challenge with 100 LD50 of the virulent wild-type (wt) strain. Immunization with either C/2/2 or H/2/26 did not confer protection. The three ts mutants induced the production of local IgA after i.g. administration regardless of their protective capacity. PMID- 8401429 TI - Stimulation of rat spleen cells by staphylococcal enterotoxins. AB - There is much interest in staphylococcal enterotoxins as T cell mitogens in humans, mice and rabbits. Rat spleen cells were shown to proliferate in response to staphylococcal enterotoxins A and B and toxic shock syndrome toxin-1 at concentrations (5 to 500 ng ml-1) which also stimulate mouse spleen cells. The proliferative response to all these enterotoxins was inhibited by cyclosporin A, indicating the response to be predominantly that of T cells. These results indicate that the rat provides another convenient model for the analysis of T cell responses to enterotoxins. PMID- 8401430 TI - Identification and immunochemical characterization of a germ tube specific antigen of Candida albicans. AB - Germ tube specific fractions of the dimorphic pathogenic fungus Candida albicans were fractionated according to their ability to link to fibrinogen. These fibrinogen binding factors were used as immunogens to prepare monoclonal antibodies (mAbs) with BALB/c mice. Among the resulting mAbs, one (mAb 3D9.3) was shown by indirect immunofluorescence to be specific to the surface of the mycelial phase of the C. albicans species. No labelling of the cell wall of any other Candida species was observed. This morphological shape specificity was confirmed by immunoblotting where a polydispersed high molecular mass component was identified. The molecular mass varied with the extraction procedure used; over 210 kDa with EDTA-2ME treatment, and ranging from 110 to 220 kDa after Zymolyase digestion. This phase-specific epitope was sensitive to proteolysis with pronase E, proteinase K and trypsin, but not to periodate treatment. Further purification of this material would allow further development of new serodiagnostic assays that might be more specific for invasive disease than currently available tests. PMID- 8401431 TI - Heat-stable opsonins in tuberculosis and leprosy. AB - We have examined heat-stable opsonins to 4 species of gamma-irradiated mycobacteria (M. tuberculosis (H37Rv), M. avium (28A), M. scrofulaceum and M. leprae (cd 103)) in complement-depleted sera collected from Indonesian subjects with tuberculosis (106 patients),-leprosy (24 patients) and controls (40 hospital workers and 41 factory workers) indirectly by microtitre plate chemiluminescence (CL) assay and compared the results with antibody levels. The results indicate that there is a wide range of opsonic capacity for mycobacteria in complement depleted sera. There was a poor correlation between the opsonic capacity as measured by CL and the anti-mycobacterial antibody content of sera measured by ELISA, suggesting that anti-mycobacterial antibody has little influence on the uptake of mycobacteria. However, a non-specific heat-stable opsonin appears to be present in some sera. Conversely, some sera from tuberculosis or leprosy patients suppress the production of reactive oxygen species from normal phagocytes in vitro when stimulated with M. tuberculosis. The relevance of this inhibition and the presence of heat-stable opsonins to the pathogenesis of tuberculosis have yet to be determined, but it is possible that the presence of opsonins may inhibit dissemination of tubercle bacilli to other organs. PMID- 8401432 TI - Vaccination strategies against intracellular microbes. PMID- 8401433 TI - Under doctor's orders in the United States. PMID- 8401434 TI - Matrons-in-waiting. PMID- 8401435 TI - Keeping in touch. PMID- 8401436 TI - Beating the system with a computer. PMID- 8401437 TI - Please call the dentist. PMID- 8401438 TI - Continence consumer test. PMID- 8401439 TI - Breaking the silence. PMID- 8401440 TI - On course for success. PMID- 8401441 TI - Eastern promise. PMID- 8401442 TI - As I was saying.... PMID- 8401443 TI - Not only for art's sake. PMID- 8401444 TI - Scottish matters. PMID- 8401445 TI - Herpes zoster (shingles). PMID- 8401446 TI - Fox in charge of the chicken coop. PMID- 8401447 TI - Synthesis of enantiomerically pure alpha-[14C]methyl-L-tryptophan. AB - A practical method for the preparation of large amounts of enantiomerically pure alpha-[14C]methyl-L-tryptophan using the enzymatic resolution of the corresponding D,L-methyl ester is reported. The radiolabelled alpha-methyl group was introduced using the alpha-methylation of the lithium enolate of the Schiff base of L-tryptophan methyl ester. Hydrolysis of the Schiff base with 1N HCl provided the D,L-methyl ester of alpha-[14C]methyl tryptophan. Enantioselective enzymatic hydrolysis of the L-methyl ester by alpha-chymotrypsin gave the enantiomerically pure alpha-[14C]methyl-L-tryptophan. The overall yield of this preparation was 33%. PMID- 8401448 TI - Effects of preparation methods of biomedical samples on PIXE measurement sensitivity. AB - Instrumental sample preparation techniques for PIXE measurements of human blood serum and bovine muscle were investigated. The techniques studied were direct measurement of a drop, drying on a foil and on an Al cup, lyophilization on a foil, pellet pressing of dried material, microtome sectioning and biopsy preparation. The same serum and muscle were used in each procedure allowing the comparison of detection limits between different methods. The most beneficial technique for both samples was lyophilization and pellet pressing. PMID- 8401449 TI - A simple method to determine the immunoreactivity of radiolabelled monoclonal antibodies to the TAG-72 antigen. AB - A simple method has been developed for determining the immunoreactivity of radiolabelled monoclonal antibodies to the TAG-72 antigen. The method involves binding of a constant small amount of the antibody to increasing concentrations of bovine submaxillary mucin. The immunoreactive fraction (IRF) is then determined by linear extrapolation of binding to infinite antigen excess. Using this assay, the IRF of radioiodinated anti-TAG-72 antibodies ranged from 0.22 0.48. PMID- 8401450 TI - Automated production of oxygen-15 labeled butanol for PET measurement of regional cerebral blood flow. AB - Because [15O]butanol is the radiopharmaceutical of choice for PET studies of cerebral perfusion and neurological activation, we have developed a microprocessor-controlled radiosynthetic system for the preparation of this radiotracer in up to ten batches at a time. An IBM-compatible minicomputer was programmed to direct the reaction of molecular [15O]oxygen with tri-(n butyl)borane bound to alumina, followed by purification of product [15O]butanol via solid phase extraction with C18 Sep-Paks and sterile filtration. Routine batch yields of over 150 mCi were achieved with a preparation turn-around time of 6.0 min. The final product had high radiochemical purity, low chemical impurity, and was sterile and apyrogenic. This radiopharmaceutical production system is reliable and suitable for tracer production in clinical PET imaging centers. PMID- 8401451 TI - Labelled polycyanoacrylate nanoparticles for human in vivo use. AB - Isobutyl and isohexyl cyanoacrylate nanoparticles are used as drug carriers, particularly for some anti-cancer drugs. Body distribution as well as pharmacokinetics have been well studied in animal and partially in man. Labelling of the monomer itself or of the carried drug with beta-emitters allowed such studies. In man, however, organ distribution and uptake could easily be done and followed by means of scintigraphy (imaging) techniques if one could achieve nanoparticle labelling with gamma-emitting isotopes. We have developed labelling methods able to supply such carriers using gamma-emitters like radioactive iodine (125I or 131I), indium or technetium. We used DTPA as a spacer in order to fix the last two isotopes. This would mean that any other gamma-emitting cation can theoretically be tried pending on its ability to be chelated by DTPA. The preparations were obtained with high labelling yields, usually > 80% and were relatively stable in human plasma over the whole period of investigation. 111In and 99mTc labelled forms have been administered to rabbit and then to man with 60 75% accumulation in the reticulo-endothelial system. PMID- 8401452 TI - The 'states versus weights' dilemma in quality of life measurement. PMID- 8401453 TI - Testing the validity of the Euroqol and comparing it with the SF-36 health survey questionnaire. AB - There is an interest in the consequences of deriving a single index measure of health for validity and sensitivity. This paper presents the results of testing a recent example of a general health measure designed to derive a single index, the Euroqol (EQ), and presents a comparison with a new, influential profile measure, the Short Form 36 (SF-36) Health Survey Instrument. The EQ and an anglicised version of the SF-36 health survey, both designed for self-completion, were included in a postal survey of a random sample of 1980 patients from two practice lists in Sheffield, UK. The response rate for the EQ questionnaire was 83%, and the rate of completion over 95%. Evidence was found for the construct validity of the EQ dimension responses and the derived total EQ health score in terms of distinguishing between groups with expected health differences. Considerable agreement was found between EQ responses and the total EQ score, and the UK SF-36 profile scores. There was substantial evidence of EQ being less sensitive at the ceiling (i.e. low levels of perceived ill-health) and throughout the range of health states. A recent restructuring of the EQ, may help overcome some of the problems with the physical dimensions by reducing their skewness. PMID- 8401454 TI - Measurement of dyspnoea in chronic obstructive pulmonary disease. AB - This paper reviews the assessment of shortness of breath in chronic obstructive pulmonary disease (COPD). The validity criteria for evaluating measures of dyspnoea are discussed and a description and critique of current measures of shortness of breath are offered. Across studies, dyspnoea measures are moderately correlated with pulmonary function (e.g. FEV1.0 and FVC), psychological function, and walking tests (6 min walk). In addition, dyspnoea measures tend to be correlated with one another. The need for standardisation of dyspnoea measures for research and clinical practice is identified as an important objective for future work. PMID- 8401455 TI - The impact of generalized malignant melanoma on quality of life evaluated by the EORTC questionnaire technique. AB - Eighty-nine patients with generalized malignant melanoma, 63% men and median age 53 years, were included in a longitudinal quality of life (QOL) study before the start of chemotherapy. QOL was assessed by the EORTC core questionnaire technique (QLQ-C36), a study-specific melanoma (MM) module and the Hospital Anxiety and Depression (HAD) scale. The questionnaires displayed good psychometric qualities and were able to document the florid symptomatology of disseminated melanoma. They were well accepted by the patients. Before treatment the patients reported a relatively low symptom burden, good physical and social functioning, moderate psychological distress and a high overall QOL rating during the past week. Fatigue and pain were the most frequent symptoms reported. The QOL measurement differentiated between subgroups of patients differing in performance status and the tumour burden. We conclude that the EORTC questionnaire technique is feasible and clinically relevant in generalized malignant melanoma patients. PMID- 8401456 TI - Compliance with the medical regimen and partner's quality of life after heart transplantation. AB - The authors investigated 40 heart transplant recipients and their partners to determine both the partner's quality of life upon transplantation and the experiences of both patient and partner with compliance with the medical regimen. Data on sleep disturbances, social isolation, emotional reactions, depression, anxiety, partner's apprehension, social support and compliance (regarding behaviour and emotional experience) were obtained approximately 21 months after transplantation. Compared to related study groups, partners did not experience more problems in sleep, social isolation, emotional reactions, depression and anxiety. Patients overestimated the apprehension of their partners significantly (p < 0.0001). Generally speaking, with the exception of three items relating to eating fish, canned food and forgetfulness in medicine intake, patients and partners agreed with respect to actual compliance behaviour. Lowest compliance concerned regular physical exercise: 28%. Both patient and partner insisted that they had scarcely any emotional problem with the regimen. Further systematic research is needed to bring to light factors that affect compliance as well as adequate methods to bring about an improvement therein. PMID- 8401457 TI - Caregivers of persons with stroke: their physical and emotional wellbeing. AB - This descriptive study was undertaken as a pilot test for several questionnaires designed to examine the well-being of caregivers of persons with stroke. This predominantly female sample was found to have moderately few physical symptoms, but to be in considerable emotional distress. Nearly half the sample had anxiety and depression scores above the level identified as suspicious for clinically relevant distress as indexed by the Symptom Questionnaire. Anger was also a salient finding: 40% of those questioned scored above the cutoff level for hostility and several caregivers expressed anger verbally during data collection. The degree of psychological distress in these individuals is of importance to all health care professionals and deserves increased attention, both from the perspective of the researcher and from the perspective of the clinician. PMID- 8401458 TI - Interpretation of quality of life changes. AB - The clinical significance of quality of life changes has little to do with the QOL as a measure, but is rather a reflection on the novelty of the measures and researcher's inexperience in their use. This article discusses possible ways of evaluating quality of life measures in terms of patient-clinician interactions and how the clinician can assess the importance of these changes of QOL in terms of treatment and management of disease. PMID- 8401459 TI - A sharper focus for the New York Academy of Medicine and its journal. PMID- 8401460 TI - Health care reform in Minnesota: lessons for the country. PMID- 8401461 TI - Towards an urban HIV policy. PMID- 8401462 TI - The Bulletin--a new focus, a new look. PMID- 8401463 TI - Reframing the debate: toward effective treatment for inner city drug-abusing mothers. PMID- 8401464 TI - Rational health care reform? PMID- 8401465 TI - New York City's Communicare concept. PMID- 8401466 TI - Municipal hospitals in New York City--a review of the Report of the Commission to Review the Health and Hospitals Corporation. PMID- 8401467 TI - Prenatal care. Appointment study: a survey by the Infant Mortality Work Group of the Mayor's Advisory Council on Child Health. AB - This biopsy of the prenatal care system provides plain evidence of weaknesses within the infrastructure and details the inconveniences New York City women who rely on public services must face. The unwillingness of some of the municipal clinics to accept pregnancy test results from their sister hospitals or the Department of Health's free-pregnancy testing program exemplifies one of the bureaucratic barriers in the system, and one which could be rectified easily given sufficient determination. Other bureaucratic barriers, such as the inability to schedule an appointment unless a woman is registered at the clinic, suggest the need for a media campaign urging women of reproductive age to register with a health care provider before becoming pregnant. The solution to the language barrier confronted by Spanish women requires a commitment to hiring bilingual clerical staff. Lastly, the problems of being placed "on-hold" or frequent busy signals and the high number of clinic visits made before seeing a physician can only be ameliorated by an increase in funding for prenatal care clinic staffing, for support staff and professionals alike. The survey, conducted over the course of 2 weeks with available city staff and equipment, is a relatively inexpensive and effective method for evaluating the prenatal care system and should be repeated in order to document systematically the anecdotal reports shared by clinic administration and clinic patients alike. PMID- 8401468 TI - Urban health data. Spending on child health services in New York City. PMID- 8401469 TI - Comparative effects of selected azole compounds on trophic and cystic stages of Acanthamoeba polyphaga. PMID- 8401470 TI - Kinetics of expression of two major Plasmodium berghei antigens in the mosquito vector, Anopheles stephensi. AB - Expression of a 21 kDa determinant (Pbs21), first detected on the surface of ookinetes, and of the circumsporozoite protein (CSP) was studied by immunofluorescence and Western blots during the developmental cycle of Plasmodium berghei in the mosquito Anopheles stephensi. The expression of Pbs21 was predominantly localised on the ookinete surface one day after the infectious blood meal, and thereafter reactivity declined to a minimum on days 2 and 3, the time of onset of oocyst development. A gradual increase in fluorescence was observed on the oocysts from day 6 that was retained until day 17 post-infection. In contrast, sporozoites released from oocysts or salivary glands showed little or no antibody labelling with anti-Pbs21. Circumsporozoite protein was not detectable in any midgut preparations until 5-6 days after feeding, when reactivity was observed against immature oocysts. Expression then continued and increased throughout oocyst and sporozoite development. Western blots confirmed that Pbs21 was expressed minimally during the oocyst development but was not detectable in sporozoites. Co-localisation of anti-Pbs21 and anti-CSP monoclonal antibodies to the 50 kDa and 60 kDa bands in Western blots of sporozoite suggests immunological cross-reactivity between the CSP and the anti-21 kDa antibodies. PMID- 8401471 TI - Distribution of macromolecular methylations in promastigotes of Leishmania donovani and impact of sinefungin. AB - Sinefungin, an antifungal and antiparasitic nucleoside antibiotic, is a very potent antileishmanial agent both in vitro and in vivo. This molecule, structurally related to S-adenosylmethionine, is a good competitive inhibitor of methyltransferases in vitro. The aim of this report is to analyze the impact of sinefungin on methylation pattern and the subcellular localisation of methyl groups and various methylases in promastigotes of Leishmania donovani. We have shown the presence of various methylated macromolecules in different subcellular fractions, with somewhat higher concentration in membrane fraction. In vitro, sinefungin inhibits the three main protein methylases, but in cells cultured in its presence the protein carboxylmethylations are specifically inhibited. PMID- 8401472 TI - Hydrolase compartmentalization limits rate of digestion in Acanthamoeba. AB - The kinetics of lysosomal enzyme acquisition by newly formed phagosomes was studied by following the rate of digestion of radiolabeled yeast fed to Acanthamoeba. The distribution of hydrolases among phagosomes was assessed by electron microscopic acid phosphatase cytochemistry and by measurement of three glycosidases in isolated early and late phagosomes. The results show that compartmentalization of hydrolases limit the digestion of large phagocytic loads. The hydrolases appear to be sequestered into the early phagosomes and not to be distributed either by small vesicle transport or phagosome-phagosome fusion to those formed later. We infer from these results that newly internalized surface membrane in phagosomes is not rapidly randomized with internal pools, but is recycled to the surface as a function of the digestive process. PMID- 8401473 TI - Amylopectin synthase of Eimeria tenella: identification and kinetic characterization. AB - A soluble enzyme amylopectin synthase (UDP-glucose-alpha 1,4-glucan alpha-4 glucosyltransferase) which transfers glucose from uridine 5'-diphosphate glucose (UDP-glucose) to a primer to form alpha-1,4-glucosyl linkages has been identified in the extracts of unsporulated oocysts of Eimeria tenella. UDP-glucose and not ADP-glucose was the most active glucosyl donor. Corn amylopectin, rabbit liver glycogen, oyster glycogen and corn starch served as primers; the latter two were less efficient. The enzyme has an apparent pH optimum of 7.5 and exhibited typical Michaelis-Menten kinetics with dependence on both the primer and substrate concentrations. The Michaelis constants (Km), with respect to UDP glucose, was 0.5 mM; and 0.25 mg/ml and 1.25 mg/ml with respect to amylopectin and rabbit liver glycogen. The product formed by the reaction was predominantly a glucan containing alpha-1,4 linkages. The specificity of the enzyme suggests that this enzyme is similar to glycogen synthase in eukaryotes and has been designated as amylopectin synthase (UDP-glucose-alpha-1,4-glucosetransferase EC 2.4.1.11). PMID- 8401474 TI - The evolution of the Vahlkampfiidae as deduced from 16S-like ribosomal RNA analysis. AB - The amoebae, a phenotypically diverse, paraphyletic group of protists, have been largely neglected by molecular phylogeneticists. To better understand the evolution of amoebae, we sequenced and analyzed the 16S-like ribosomal RNA genes of three vahlkampfiid amoebae: Paratetramitus jugosus, Tetramitus rostratus and Vahlkampfia lobospinosa. The Vahlkampfiidae lineage is monophyletic, branches early along the eukaryotic line of descent, and is not a close relative of the multicellular amoebae that also reversibly transform from amoebae to flagellates. PMID- 8401475 TI - Fine structure and cytochemistry of Tritrichomonas foetus and rat neutrophil interaction. AB - The fine structure of normal and antibody-coated Tritrichomonas foetus cells and their interaction with rat peritoneal neutrophils was studied. Peritoneal neutrophils were obtained by glycogen stimulation. The neutrophils readily associated with and killed the parasites, which were subsequently ingested. The process involved activation of the respiratory burst, as demonstrated by the use of cytochemical methods. Images were obtained indicating that binding of parasites to the neutrophil surface triggers an exocytic response with release of oxygen-derived products. Cytochemical localization of acid phosphatase and peroxidase activities showed that leukocyte granules fused with the parasite containing phagocytic vacuoles. We also showed the cytochemical localization of alkaline phosphatase in the parasite-neutrophil interaction. PMID- 8401476 TI - Purine nucleoside and nucleobase cell membrane transport in Giardia lamblia. AB - Giardia lamblia is dependent on the salvage of preformed purines and pyrimidines. This study investigated purine nucleoside and nucleobase transport utilizing rapid uptake determinations. Nucleoside substrate/velocity curves exhibited the hyperbolic kinetics of a saturable carrier-mediated system. Deoxynucleosides exhibited a much lower affinity for the transporter. Inhibition studies confirmed the relative carrier affinities of these ribonucleosides and deoxyribonucleosides. The nucleobase adenine did not exhibit saturation kinetics at a comparable substrate range, and did not inhibit nucleoside transport. Dipyridamole markedly inhibited nucleoside but not nucleobase transport, confirming the separate entry pathways. When cells were depleted of ATP, the velocity of nucleoside and nucleobase transport was unchanged, indicating that it is a non-energy-dependent process. Three nucleoside analogs, formycin A, adenine arabinoside and 7-deazaadenosine, were studied. Transport kinetics ranged widely among this group and could not completely account for their cytotoxic effect. When the apparent Km and Vmax of the nucleosides were compared, an approximately linear relationship (r2 = 0.95) was noted. This suggests that a high affinity of the nucleoside permease for the substrate retards disassociation of the substrate carrier complex, slowing net influx. PMID- 8401477 TI - Biochemical analysis of a mutant Tetrahymena lacking outer dynein arms. AB - Tetrahymena thermophila mutants homozygous for the oad mutation become nonmotile when grown at the restrictive temperature, and axonemes isolated from nonmotile mutants lack approximately 90% of their outer dynein arms. Electrophoretic analyses of axonemes isolated from nonmotile mutants (oad axonemes) indicate they contain significantly fewer of the 22 S dynein heavy chains that axonemes isolated from wild-type cells (wild-type axonemes) contain. The 22 S dynein heavy chains that remain in axonemes isolated from nonmotile, oad mutants are assembled into 22 S dynein particles that exhibit wild-type levels of ATPase activity. Two dimensional gel electrophoresis of oad axonemes show that they are deficient in no proteins other than those proteins thought to be components of 22 S dynein. This report is the first formal proof that outer dynein arms in Tetrahymena cilia are composed of 22 S dynein. PMID- 8401478 TI - Ribosomal RNA sequencing of members of the Crypthecodinium cohnii (Dinophyceae) species complex; comparison with soluble enzyme studies. AB - Sixty-five members of the Crypthecodinium cohnii species complex were analyzed for sequence differences within the D2 region of the 23S ribosomal RNA molecule. On the basis of 46 sequence differences the strains fell into 19 distinct ribosets (strains of identical sequence), some with many members. Members of four of the seven major sibling species (widespread breeding groups) were each found within single ribosets. Members of three other major sibling species were each, however, divided into two ribosets by a single sequence difference correlated with geographic separation and with previously reported electrophoretic polymorphisms of soluble enzymes within the sibling species. In addition to members of major sibling species, some ribosets include many minor sibling species (each represented by only one strain). Of 38 minor sibling species, 22 shared sequence with a major sibling species. Of these 22, 14 were identical in soluble enzymes to their related major sibling species or differed by only one of three enzymes. Other minor sibling species appear to have diverged extensively from any others in both rRNA sequence and electrophoretic profile. As a group, major sibling species differ markedly in the number of minor sibling species associated with them, suggesting differences in frequency of sexually isolating events in their past histories. These findings are discussed in the context of the previously proposed model of sympatric speciation. PMID- 8401479 TI - The dcc mutation affects ciliary length in Tetrahymena thermophila. AB - We have characterized ciliogenesis in a mutant Tetrahymena thermophila that both fails to regain motility following deciliation and that fails to complete cytokinesis. Scanning electron microscopic (SEM) observations revealed that starved deciliated cells regenerated fewer, shorter cilia at the restrictive temperature than similarly treated cells incubated at the permissive temperature. Transmission electron microscopic evaluation of isolated, regenerated cilia revealed no structural abnormalities. Incorporation of S-35 methionine was similar during ciliary regeneration at both the restrictive and permissive temperatures, indicating the mutant phenotype was not due to a simple failure in translation or transcription. Mutant cells incubated in growth medium at the restrictive temperature arrested in cytokinesis and assembled a large number of abnormally short cilia. These cells also developed irregular surface projections that were not visible on wild-type cells. These observations suggest that ciliogenesis can be initiated in growing cells as well as in starved deciliated cells but that elongation is inhibited before cilia reach full length. The mutation was named dcc for defective in ciliogenesis and cytokinesis. PMID- 8401480 TI - Some aspects of the humoral and neutrophil functions in post-comatose unawareness patients. PMID- 8401481 TI - Ethics in brain injury rehabilitation: existential choices among western cultural beliefs. AB - The following issues in the practice of brain injury rehabilitation are explored: (1) validity of support for efficacy based on experience in clinical practice, (2) validity of support for efficacy based on research, (3) consumer protection, (4) qualifications and regulation of individual providers, (5) regulation of programme development and marketing. Ethical responses to these issues from each of three cultural belief systems (humanism, science and self-interest) are examined from a metaphilosophical perspective based on contemporary cognitive psychology and on philosophies of social constructionism and existentialism. PMID- 8401482 TI - Some aspects of the humoral and neutrophil functions in post-comatose nawareness patients. AB - Post-comatose unawareness (PCU) is one of the possible outcomes of severe brain injury. Patients with severe brain injury have an increased susceptibility to severe nosocomial infections for multifactorial reasons, including immune suppression at different levels. We studied different immunological aspects in 11 PCU patients. Impaired humoral immunity was found in 27% of them. Two patients had decreased haemolytic activity of the classical complement pathway, associated with decreased levels of the components C1q, C1r and C4. Another patient had very low levels of IgG2 and IgG4. The neutrophil killing activity was impaired in these three patients, but was completely restored with the addition of a heterologous serum, suggesting a humoral defect. Neutrophils showed normal chemotaxis and random migration, and the superoxide anion release by neutrophils was also found to be normal. Understanding the immunological events in PCU patients contributes to a better and more intensive therapeutic approach, which might accelerate the rehabilitation of these patients. PMID- 8401483 TI - Abstract thinking following severe traumatic brain injury. AB - Abstract abilities were studied in a sample of 34 individuals with severe TBI and a control group. The results indicate that TBI interferes with performance on tests requiring individuals to process information into new categories. There appears to be a dissociation between verbal abstract abilities and visual perceptual abstract abilities. There is evidence that Goldstein and Sheerer's [1] postulate of a general 'abstract attitude' was at least partially correct. This attitude does not appear to be related to a general verbal ideational process, as dysphasic subjects were only deficient on a purely verbal abstract task. PMID- 8401484 TI - Limitations of neurological assessment in mild head injury. AB - The authors have investigated two commonly used methods of assessing neurological status in patients with mild head injury to determine whether they can predict intracranial damage. Of 686 such patients with cranial computed tomography (CT) scans, scan results were recorded, along with total and motor components of the Glasgow Coma Scale (GCS) and the Reaction Level Scale (RLS85). Despite relatively normal admission neurological examinations, 127 of the 689 patients (18.4%) had intracranial lesions, and 38 (5.5%) required surgery. There was no significant difference in distribution of the GCS in patients with and without intracranial lesions. The RLS85 was superior to the GCS in predicting intracranial pathology, and a significant association between RLS85 and lesions on CT scanning was noted. However, even this test was normal in 19 patients found to have intracranial pathology, including nine who required surgery. The authors conclude that a normal or near-normal mental status examination in a head-injured patient on arrival at the emergency room is inadequate to exclude a potentially serious intracranial lesion. It is unlikely that further refinements in the clinical evaluation will result in diagnostic accuracy comparable with that of CT scanning. Accordingly, we recommend that any patient who has suffered a loss of consciousness or amnesia following head injury have an urgent cranial CT scan. PMID- 8401485 TI - Memory functioning after traumatic brain injury in children. AB - Immediate and 45-minute delayed recall of a paragraph-length story and of a complex geometric figure were investigated in a sample of 30 children with traumatic brain injury. There was no significant difference between children with mild to moderate injuries and children with severe injuries with regard to general level of verbal recall. However, there was a trend for children with mild to moderate injuries to have better recall of visual information than children with severe injuries. Recall of verbally presented information deteriorated significantly over the 45-minute delay, regardless of injury severity. No such deterioration was found for recall of visually presented information. Clinical and research implications are discussed. PMID- 8401486 TI - Incidence and treatment of visual dysfunction in traumatic brain injury. AB - The incidence of visual dysfunction and effectiveness of visual exercises in acute traumatically brain injured inpatients in a rehabilitation programme were studied. Vision evaluation norms were established on 23 hospital staff. The evaluation was then administered to 51 inpatients within days after admission. An additional 21 patients were unable to participate, usually due to decreased cognition or agitation. Thirty of 51 (59%) scored impaired in one or more of the following: pursuits, saccades, ocular posturing, stereopsis, extra-ocular movements, and near/far eso-exotropia. For patients having dysfunction in pursuits or saccades, a 2-week baseline was followed by vision exercises. During the baseline interval patients were evaluated by an optometrist to verify therapists' findings. Six patients who participated in several weeks of treatment were evaluated at 2-week intervals by an independent rater. Progress is graphically illustrated. Conclusions were that the suitability of an inpatient vision programme, from our experience, is questionable. However, an initial evaluation proved valuable for informing staff of patients' visual status and for referral to an optometrist/ophthalmologist for further treatment. PMID- 8401487 TI - Survey of case manager training needs in traumatic brain injury. AB - The results of 138 surveys assessing the training needs of case managers in the area of traumatic brain injury were analysed. Sixty-six per cent of the respondents were case managers working primarily with adults in a variety of work settings. Respondents rated working with families, vocational rehabilitation, and community re-entry as the top three areas for training. Mild traumatic brain injury, paediatric traumatic brain injury, neuropsychological assessment, and medicolegal aspects received the highest rankings by case managers with more than 8 years experience. The need for certification of case managers in specialty areas was supported by 83% of all respondents. PMID- 8401488 TI - Potentially toxic serum concentrations of desipramine after discontinuation of valproic acid. AB - Pharmacological interventions in the treatment of various cognitive, behavioural and neurological problems after brain injury often may involve combinations of medications from various drug classes. This carries the implication of potentially new or previously underreported drug interactions. A case report is presented in which a commonly used anticonvulsant drug, valproic acid, and a commonly used antidepressant, desipramine, interacted in such a manner as to cause potentially toxic serum concentrations of desipramine. This case demonstrates the important point that it is not simply the addition of one drug to another that may cause interaction, but the withdrawal of a particular drug which may then adversely impact the remaining drug regimen. PMID- 8401489 TI - The leading role of STSs in genome mapping. PMID- 8401490 TI - The interleukin-2 receptor gamma chain maps to Xq13.1 and is mutated in X-linked severe combined immunodeficiency, SCIDX1. AB - The gene encoding the gamma chain of the lymphocyte interleukin-2 receptor has been cloned and shown to be required to associate with the beta chain in order for IL-2 internalization and cell activation to occur (1). We considered this gene, IL2RG, a candidate for the X-linked form of severe combined immunodeficiency at the SCIDX1 locus, in which affected males have impaired lymphocyte development. Using fluorescence in situ hybridization and PCR amplification of somatic cell hybrid DNAs, we mapped IL2RG to human Xq13.1, a location within the SCIDX1 critical region established by linkage analysis. The 4.2 kb IL2RG gene was sequenced, and its genomic organization was elucidated. Seven of 19 transformed B-lymphocyte cell lines with independent SCIDX1 mutations had absent or minimal IL2RG mRNA. Unique point mutations were documented to be specifically associated with the disease and the carrier state in four unrelated affected males and their family members: one in a boy with no detectable IL2RG mRNA, in which the mutation ablated a splice donor site; one causing premature chain termination; and two causing distinct amino acid changes. The demonstration of impaired IL2RG mRNA expression in males with X-linked SCID and of unique point mutations in SCIDX1 pedigrees constitutes powerful evidence that the SCIDX1 gene is IL2RG. Noguchi et al. (2) have independently published IL2RG mapping to Xq13 and discovery of mutations in three affected males. The specific pathogenesis of IL2RG mutations and approaches to gene therapy can now be addressed in the X linked form of SCID. PMID- 8401491 TI - 2.6 Mb YAC contig of the human X inactivation center region in Xq13: physical linkage of the RPS4X, PHKA1, XIST and DXS128E genes. AB - X chromosome inactivation is a mechanism of dosage compensation that regulates the expression of mammalian X-linked genes between XY males and XX females. This phenomenon is cis-acting, clonally heritable, and requires the presence of an X inactivation center (XIC). In our attempts to characterize this phenomenon, we have focused on the physical organization of the human XIC localized to Xq13. From previous studies, we had determined that the candidate XIC interval contained two loci (DXS128 and XIST) and was bound by the breakpoints of two structurally abnormal inactivated X chromosomes, a t(X;14) and an idic(Xp). Here we present a refined mapping of the XIC-containing region using the breakpoint of a late replicating rearranged X (rea(X)), and the initial characterization of a set of 40 yeast artificial chromosomes (YACs) derived from the XIC-containing region. These YACs form a 2.6 Mb contig which completely covers the XIC, and physically links the RPS4X, PHKA1, XIST, and DXS128E genes, as well as a laminin receptor pseudogene (LAMRP4). Furthermore, we have determined the relative orientations of these four genes, and have derived a restriction map of the region using the rare cutter enzymes BssHII, EagI, MluI, NruI, SalI, SfiI, SstII (or SacII), and NotI. We have identified at least 9 CpG-rich islands within this region, and have discovered a large (approximately 125 kb) inverted duplication proximal to the XIC based on symmetrical restriction patterns and homologous probes. We estimate the maximum size of the XIC-containing interval to be between 680 kb and 1200 kb, based on the localization of the breakpoints of the rearranged X chromosomes mentioned above. This lays the groundwork for the further characterization of the XIC region and the isolation of other expressed sequences therefrom. PMID- 8401492 TI - Generation of band-specific painting probes from a single microdissected chromosome. AB - We have developed a modified strategy for the generation of regional probes for human chromosomes by microdissection and degenerate oligonucleotide primed PCR. This modification dramatically increases the efficiency of amplification by pretreatment of the dissected chromatin with topoisomerase I (Topo I) before PCR. This protocol has enabled us to construct region-specific probes for fluorescence in situ hybridization (FISH) from a single microdissected chromosome. Results are presented which convincingly demonstrate that this new method generates high intensity region-specific FISH probes, while at the same time significantly decreasing the time-consuming and labor-intensive aspects of microdissection. The reduction of the number of copies required to generate a useful probe also significantly decreases the risk of contamination during the microdissection process. We believe this advance will allow microdissection to be more widely used in the cytogenetic analysis of chromosome rearrangements in both cancer and hereditary diseases. In addition, this method now makes it possible to construct a series of non-overlapping band-specific DNA microclone libraries to provide complete coverage of individual chromosomes for physical mapping. PMID- 8401493 TI - Mutation of human short tandem repeats. AB - A total of 20,000 parent-offspring transfers of alleles were examined through the genotyping within 40 CEPH reference families of 28 short tandem repeat polymorphisms (STRPs) located on chromosome 19. Forty-seven initial mutation events were detected in the STRPs using DNA from transformed lymphoblastoid cell lines, but less than half (39%) could be verified using DNA from untransformed cells. None of the cases where three alleles were observed in a single individual could be verified using DNA from untransformed cells. The average mutation rate for the chromosome 19 STRPs after correction for events which would not be detectable as Mendelian errors was 1.2 x 10(-3) per locus per gamete per generation. This rate may have been inflated by somatic as opposed to germline events. Observed mutation rates for individual STRPs ranged from 0 to 8 x 10(-3). The average mutation rate for tetranucleotide STRPs was nearly four times higher than the average rate for dinucleotide STRPs. For determination of the mode of mutation, events involving STRPs on other chromosomes were also examined. Of the events which were verified using DNA from untransformed lymphocytes or which were likely among those for which DNA from untransformed cells was not available: none were located at the sites of meiotic recombination, 91% involved the gain or loss of a single repeat unit, and 15 occurred in the male germline compared to 4 in the female germline (p = 0.01). PMID- 8401494 TI - Digital DNA typing at a second hypervariable locus by minisatellite variant repeat mapping. AB - Minisatellite variant repeat unit mapping by PCR (MVR-PCR) assays the interspersion pattern of variant repeat units along minisatellite alleles. Mapping such internal variation in the highly polymorphic minisatellite MS31A (locus D7S21), reveals extreme levels of allelic variability, far in excess of that detectable by allele length analysis. Flanking base substitutional polymorphisms have enabled the 5' structure of large numbers of MS31A alleles to be derived from genomic DNA by allele-specific MVR-PCR. More than 100 alleles have now been mapped and all are different. Several alleles show related internal structures and some of these provide evidence of polarity in allelic variation reminiscent of that seen at two other hypervariable minisatellites, D1S8 (MS32) and D16S309 (MS205). We also describe the diploid digital coding of MS31A, including the simultaneous coding of MS31A and a second locus, MS32, by duplex MVR-PCR, which greatly enhances the potential forensic applications of this technique. PMID- 8401495 TI - Allelic diversity at minisatellite MS205 (D16S309): evidence for polarized variability. AB - We have determined the allelic structures on 106 Caucasian chromosomes at the minisatellite locus MS205 (D16S309). In addition to the internal structures deduced by minisatellite variant repeat (MVR) mapping, the genotypes at six flanking substitutional polymorphic sites have been analysed. The minisatellite structures show a polarity of variation at MS205, with most observed variation due to differences at one extremity of the tandem repeat array. This locus, therefore, provides a further example of polarity of variation at human minisatellites. Analysis of haplotypes at flanking polymorphic sites suggests that there may be a higher frequency of exchange near the highly unstable end of the minisatellite, and thus that exchanges of flanking polymorphisms may be due to co-conversion or recombination events occurring during unequal exchanges between minisatellite alleles. PMID- 8401496 TI - Fine structure of the human FMR1 gene. AB - The fragile X syndrome is due to a CGG triplet expansion in the first exon of FMR1, resulting in hypermethylation and extinction of gene expression. To further our understanding of the gene's involvement in the syndrome, we report the physical structure of this locus. A high resolution restriction map of the FRAX(A) locus has been prepared encompassing approximately 50 kb. Using exon-exon PCR and restriction analysis, the FMR1 gene has been determined to consist of 17 exons spanning 38 kb of Xq27.3. Each intron-exon boundary has been sequenced. In general, the splice donors and acceptors located in the 5' portion of the gene demonstrate greater adherence to consensus than those in the 3' end, providing a possible explanation for the finding of alternative splicing in FMR1. The elucidation of the exon composition of the FMR1 gene and its flanking region will enhance detection of coding sequence mutations possible in fragile X phenocopy individuals. PMID- 8401497 TI - A contig of non-chimaeric YACs containing the spinal muscular atrophy gene in 5q13. AB - We have constructed a contig of non-chimaeric yeast artificial chromosomes (YACs) across the candidate region for childhood autosomal recessive spinal muscular atrophy (SMA) in 5q13. A novel microsatellite reduces the candidate region to approximately 400kb of DNA distal to D5S435. The candidate region contains blocks of chromosome 5 specific repeats which have copies on 5p as well as elsewhere on 5q. Restriction mapping of the YACs reveals at least one CpG island in the SMA gene region. The YAC maps indicate that the contig contains minimal rearrangements or deletions. The data show the value of screening several YAC libraries simultaneously in order to construct a set of overlapping sequences suitable for candidate gene searches and direct genomic sequencing. PMID- 8401498 TI - High resolution physical map of the region surrounding the spinal muscular atrophy gene. AB - Spinal muscular atrophy (SMA) is the second most common lethal, autosomal recessive disease in Caucasians, second only to cystic fibrosis. In an effort to identify the causative gene in SMA, we have used radiation hybrid (RH) mapping to prepare a high resolution physical map of the proximal region of chromosome 5 (5q11-13) which contains the SMA gene. The map of the SMA region, which spans approximately 4 Mb, contains 19 loci including 9 polymorphic DNA markers, 8 monomorphic sequence tagged sites (STS) and two genes. Based upon the RH map the two polymorphic loci which most closely flank the SMA locus were estimated to be separated by approximately 750 kb. Using two different directional cloning schemes, several new clones between the genetic markers which most closely flank SMA were isolated. These new clones within the SMA candidate region, together with cosmid clones prepared from one RH hybrid which retains an approximately 1 Mb segment spanning the SMA region as its only human DNA, will greatly facilitate efforts to identify the gene for SMA. In addition, analysis of cloned DNA segments from within the SMA candidate region has identified the presence of a novel, chromosome 5-specific, low copy repeated sequence which is distributed throughout the region containing the SMA gene as well as in at least four other regions of chromosome 5. Whether or not these novel repeated sequences throughout the SMA region are involved in the disease remains to be determined. PMID- 8401499 TI - Detection of aneuploidy and chromosomal mosaicism in human embryos during preimplantation sex determination by fluorescent in situ hybridisation, (FISH). AB - Five couples at risk of producing offspring with X-linked recessive disease underwent in vitro fertilisation with a view to preimplantation determination of embryo sex and selective transfer of females. On day three postinsemination, one or two blastomeres were removed by embryo biopsy, and used for dual fluorescent in situ hybridisation with X and Y chromosome-specific DNA probes. In two cases, two female embryos were transferred and one pregnancy, (sex confirmed), is ongoing at 19 weeks. All eight embryos from one couple were of such poor quality that diagnosis was possible in one only. In the remaining two cases no embryos were transferred due to the detection of an abnormal number of X chromosome signals. Investigation of the biopsied embryos that were not transferred revealed evidence of mitotic non-disjunction in one and of complete X monosomy in a second. A surviving fetus with this latter constitution would have developed Turner syndrome and would also have been at high risk of X-linked disease. The use of fluorescent in situ hybridisation rather than the polymerase chain reaction allowed the detection of abnormal copy numbers of X chromosomes thus preventing the transfer of potentially abnormal zygotes. PMID- 8401500 TI - Preimplantation prevention of X-linked disease: reliable and rapid sex determination of single human cells by restriction analysis of simultaneously amplified ZFX and ZFY sequences. AB - In vitro fertilization (IVF), blastomere biopsy of the 6-8 cell embryo, and single cell DNA diagnosis allows couples at risk of transmitting an X-linked or autosomal disease to start a pregnancy knowing their child will not be affected. We present a quick and reliable nested PCR strategy for sex determination at the single cell level by simultaneous amplification and subsequent restriction fragment analysis of the homologous but non-allelic ZFX and ZFY genes present on the X and Y chromosomes respectively. Amplified ZFX and ZFY sequences are of equal size and produce distinguishable HaeIII digestion products. In a randomized, blinded study of 194 individually isolated lymphoblasts, amniocytes, chorion villus cells, and blastomeres, 191 amplified successfully (98.4% sensitivity). None of the sample blanks showed any PCR product, all 90 of the karyotypically XY cells were correctly genotyped as ZFX/ZFY, all 83 of the 84 XX cells that amplified were correctly genotyped as ZFX only, and analyses of all same-embryo blastomeres were completely concordant (100% specificity). This strategy avoids a source of misdiagnosis observed in methods which detect only Y specific sequences, where amplification failure in an XY cell results in an erroneous XX diagnosis. This rapid (6 hr) and simple method of analysis, when applied to preimplantation embryo diagnosis, allows the avoidance of offspring affected with an X-linked recessive disorder by transferring only female embryos for implantation and ensuing pregnancy. PMID- 8401501 TI - Genetic mapping of the breast-ovarian cancer syndrome to a small interval on chromosome 17q12-21: exclusion of candidate genes EDH17B2 and RARA. AB - A susceptibility gene for hereditary breast-ovarian cancer, BRCA1, has been assigned by linkage analysis to chromosome 17q21. Candidate genes in this region include EDH17B2, which encodes estradiol 17 beta-hydroxysteroid dehydrogenase II (17 beta-HSD II), and RARA, the gene for retinoic acid receptor alpha. We have typed 22 breast and breast-ovarian cancer families with eight polymorphisms from the chromosome 17q12-21 region, including two in the EDH17B2 gene. Genetic recombination with the breast cancer trait excludes RARA from further consideration as a candidate gene for BRCA1. Both BRCA1 and EDH17B2 map to a 6 cM interval (between THRA1 and D17S579) and no recombination was observed between the two genes. However, direct sequencing of overlapping PCR products containing the entire EDH17B2 gene in four unrelated affected women did not uncover any sequence variation, other than previously described polymorphisms. Mutations in the EDH17B2 gene, therefore do not appear to be responsible for the hereditary breast-ovarian cancer syndrome. Single meiotic crossovers in affected women suggest that BRCA1 is flanked by the loci RARA and D17S78. PMID- 8401502 TI - A Null mutation in the vasopressin V2 receptor gene (AVPR2) associated with nephrogenic diabetes insipidus in the Hopewell kindred. AB - Congenital nephrogenic diabetes insipidus (DIR) is a rare X-linked hereditary disorder in which the renal collecting duct is unresponsive to arginine vasopressin; thus, the urine is consistently hypotonic to plasma. Recently, the association between the V2 receptor gene (AVPR2) and DIR has been proven. We have determined the gene sequence of four family members, from three generations, of a large North American family with CNDI who were originally part of the study used to formulate the Hopewell hypothesis. It had been proposed that a single DIR gene defect was introduced to North America by a member of an Ulster Scot kindred arriving on the ship Hopewell in 1761. DNA sequencing of the AVPR2 has identified a single base transversion from G-->A which changes tryptophan 71 to a stop codon in affected patients. This point mutation causes a truncation of the receptor leading to an essentially null allele. These data and other recently described mutations in the AVPR2 in North American pedigrees, descended from Ulster Scot ancestors and other origins, make the assertion of a founder effect proposed in the Hopewell hypothesis invalid. PMID- 8401503 TI - Six additional mutations in fucosidosis: three nonsense mutations and three frameshift mutations. AB - The rare lysosomal storage disease, fucosidosis results from an almost complete deficiency of alpha-L-fucosidase (EC 3.2.1.51). We have identified six new potential disease causing mutations detected by PCR amplification and sequencing of all 8 exons of the alpha-L-fucosidase gene FUCA1. (1) A C to T mutation (Q77X) in exon 1 of two Jewish-Italian siblings. This mutation was present in one allele and was found also in the mother who was of Italian origin. (2) A C to A mutation (W382X) in exon 6 in an Italian patient. This mutation was found in one allele and obliterates a unique Hphl site. (3) A C to A mutation (Y211X) in exon 3 in a Belgian patient. This mutation obliterates a unique Rsal site and was present in both alleles. (4) A homozygous single base (C) deletion in exon 2 in an Italian patient. This deletion results in a frameshift mutation (P141fs) and obliterates a unique Eael site. (5) A homozygous single base (C) deletion in exon 5 in a Portuguese patient, which also results in a frameshift mutation (S265fs). (6) A single base (A) deletion in exon 3 in a Canadian-Indian patient, which also results in a frameshift mutation (S216fs). The S216fs mutation was found in only one allele; the mutation in the other allele is not yet known. PMID- 8401504 TI - Germline deletion in a neurofibromatosis type 2 kindred inactivates the NF2 gene and a candidate meningioma locus. AB - Neurofibromatosis type 2 (NF2) is an autosomal dominant disease which predisposes to the development of schwannomas, meningiomas, ependymomas, and juvenile cataracts. The NF2 gene (NF2) has recently been isolated and maps to chromosome 22q12 between the loci D22S212 and D22S32. Deletion studies in sporadic and NF2 associated schwannomas and meningiomas, and the presence of inactivating mutations in NF2 in patients suggest that it acts as a tumor suppressor gene. A candidate meningioma gene (MEN) has also been isolated from the same interval. A new highly polymorphic (CA)n marker, D22S268, which maps very near to NF2, has allowed us to identify a kindred with three living affected individuals, where the disease is presumably caused by a large germline deletion. Fluorescence in situ hybridization and pulsed field gel electrophoresis confirm the presence of a 700kb deletion which includes the neurofilament heavy chain subunit gene locus (NEFH), D22S268, NF2 and the putative MEN gene. The absence of meningiomas in this pedigree raises doubts as to the existence of a separate MEN locus in this region. These results support the hypothesis that NF2 results from the inactivation of a tumor suppressor gene on chromosome 22q. PMID- 8401505 TI - No imprinting involved in the expression of DM-kinase mRNAs in mouse and human tissues. AB - To explain the restriction of early onset cases of myotonic dystrophy (DM) to maternal transmittance and the significant excess of male transmitters in the last asymptomatic generation, the involvement of parental effects on the autosomal dominant mode of inheritance has been suggested. Using FISH we confirmed that the DM-kinase gene is proximal to the ApoE gene on mouse chromosome 7, close to an imprinted segment. To study whether there is any firm molecular basis for the speculation that imprinting may be involved in DM we have analysed the expression of paternal and maternal alleles of the DM-kinase gene in human and mouse tissues. Length polymorphisms in the 3' non coding exons of human and mouse DM kinase genes, i.e. the variable [CTG]n repeat motif in humans and a newly identified Cn stretch variation in mice, served as tools to distinguish between allelic RNA products in various tissues. In human tissues, presence of transcripts from both parental alleles could be demonstrated by RT-PCR. In mouse, similar observations were made using a RNAse protection assay on fetal and adult muscle RNAs. We conclude that imprinting does not play a role in the expression of the DM kinase gene. PMID- 8401506 TI - Relationship between Charcot-Marie-Tooth 1A and Smith-Magenis regions. snU3 may be a candidate gene for the Smith-Magenis syndrome. AB - The juxtacentromeric region of the human chromosome 17 short arm (17p11.2-p12) contains genes involved in the Charcot-Marie-Tooth type 1A disease (CMT1A) and the Smith-Magenis syndrome (SMS). CMT1A is associated with a duplication of a short segment whereas SMS is linked to microdeletions, extending toward the centromere. We describe the construction and analysis of a 5 Mb YAC contig spanning the CMT1A duplicated segment and the distal part of four SMS microdeletions. We concluded that the YAC contig contains about 1Mb of genomic DNA which is deleted in the four SMS patients analysed. Moreover two YACs contain both STS deleted in SMS (U3) and STS duplicated in CMT1A (5H5), but the proximal breakpoint associated with the CMT1A duplication is not the same as the distal SMS breakpoint we studied. Finally we located five new STS in SMS deletion. Two of them, a microsatellite (D17S805(23)) and the gene coding for small nuclear RNA U3, have been localized in the contig we described. We may also note that snU3 is the first expressed sequence localized in an SMS deletion so far. The possible participation of this gene in the SMS phenotype is discussed. PMID- 8401507 TI - Comparative mapping of 9 human chromosome 22q loci in the laboratory mouse. AB - We present a comparative map of genes on human chromosome 22q and homologous loci in the mouse genome. Gene order in humans was established using a panel of somatic cell hybrids. Genetic maps spanning homologous segments on three mouse chromosomes were generated using an interspecific backcross. The conserved linkage between human chromosome 22 and mouse chromosome 16 includes two closely linked loci, Comt and IgI-1. The second conserved linkage involves human chromosome 22 and mouse chromosome 11 and contains two genetically and physically linked loci, Lif and Nfh. Finally, conserved synteny involving mouse chromosome 15 and human chromosome 22 spans 30 cM and contains five loci (Acr, Bzrp, Dia-1, Il2rb and Pdgfb). Loci within this conserved synteny have been sublocalized to different portions of human chromosome 22. The order of genes on mouse chromosome 15 and human chromosome 22 provides further evidence for chromosomal rearrangements within the conserved synteny that have occurred since the divergence of lineages leading to mice and humans. PMID- 8401508 TI - The major centromeric array of alphoid satellite DNA on the human Y chromosome is non-palindromic. AB - We have determined the orientation of the major centromeric alphoid array on the human Y chromosome. A PCR assay was used to analyse the vector-insert junctions of seven YAC clones previously positioned on two independent Y chromosomes. The orientation is the same at all 10 positions measured. This suggests that the alphoid array is a simple unidirectional repeat throughout and that human centromere structure thus differs from the palindromic organisation found in Schizosaccharomyces pombe. PMID- 8401509 TI - The development of sequence-tagged sites for human chromosome 4. AB - As part of our efforts to construct a high-resolution physical map of human chromosome 4, we developed a systematic approach for efficiently generating large numbers of chromosome-specific sequence-tagged sites (STSs). In this paper, we describe how rate-limiting steps in our STS development were identified and overcome, and detail our current development strategy. We present information for 822 new human chromosome 4-specific STSs, including PCR amplification conditions and subchromosomal localization data, obtained by analysis of the STS with somatic cell hybrids containing different portions of human chromosome 4. Although most STSs presented here were developed from anonymous clones whose sequences were determined in this laboratory, several STSs were developed for genes and other DNA sequences that were previously mapped to chromosome 4. Our data indicate that the availability of DNA sequence for an STS locus, in addition to the sequences of the two PCR oligonucleotides, significantly increases the transfer of that STS by allowing investigators to select new oligonucleotides best suited to the standard conditions used in their laboratories. PMID- 8401510 TI - Presence of the Mediterranean PKU mutation IVS10 in Latin America. PMID- 8401511 TI - McArdle's disease: a nonsense mutation in exon 1 of the muscle glycogen phosphorylase gene explains some but not all cases. AB - McArdle's disease is an inherited disease that results from a lack of functional muscle glycogen phosphorylase. We report here the identification of a C to T transition in exon 1 of the muscle phosphorylase gene found in all patients studied. This base pair mutation results in the substitution of a stop codon (TGA) for the codon (CGA) for Arg49 in the mature protein, and generates a novel restriction site for NlaIII. Of sixteen McArdle's patients, ten are homozygous for this mutation; the remainder are heterozygous. Additional unidentified mutations must lead to the McArdle's phenotype in the latter group of patients. PMID- 8401512 TI - X-linked recessive primary retinal dysplasia is linked to the Norrie disease locus. AB - X-linked primary retinal dysplasia (PRD) refers to an abnormal proliferation of retinal tissue causing either its neural elements or its glial tissue to form folds, giving rise to gliosis. A Jewish family of oriental origin was previously reported by Godel and Goodman, in which a total of five males suffer from different degrees of blindness. The authors postulated that the described findings are distinguished from Norrie disease, since in this case no clinical findings, other than those associated with the eyes, were noticed in the affected males. In addition, two of the carrier females exhibit minimal eye changes. We have performed linkage analysis of the family using the L1.28, p58-1 and m27 beta probes, and DXS426 and MAOB associated microsatellites. Our results map the gene responsible for the disorder between the MAOB and DXS426, m27 beta and p58-1 loci, on the short arm of the X chromosome at Xp11.3, which suggest the possibility that the same gene is responsible for both primary retinal dysplasia and Norrie disease. PMID- 8401513 TI - The high frequency of TTR M30 in familial amyloidotic polyneuropathy is not due to a founder effect. AB - Familial amyloidotic polyneuropathy (FAP) is an autosomal dominant disease due to mutations in the transthyretin (TTR) gene. Valine30-->methionine (TTR M30) is by far the most common mutation in patients with FAP. In a sample of 11 North American unrelated patients, we previously found that 6 had TTR M30. By utilizing double PASA, we could perform haplotype analysis despite the absence of DNA samples on relatives. The results indicate that at least four of the six patients with TTR M30 have different haplotypes, an observation that is surprising for North American patients in which the ostensible symptoms generally begin after the reproductive years. It is suggested that the most likely explanation is rapid selection against TTR M30 mutations by one of four possible mechanisms. PMID- 8401514 TI - A mutation in the 3' untranslated region of the factor IX gene in four families with hemophilia B. PMID- 8401515 TI - Two novel mutations causing mucopolysaccharidosis type I detected by single strand conformational analysis of the alpha-L-iduronidase gene. PMID- 8401516 TI - Acute intermittent porphyria caused by an arginine to histidine substitution (R26H) in the cofactor-binding cleft of porphobilinogen deaminase. PMID- 8401517 TI - Identification of type I collagen gene (COL1A2) mutations in nonlethal osteogenesis imperfecta. PMID- 8401518 TI - Repeat polymorphisms in human fibrillin genes on chromosome 15 (FBN1) and chromosome 5 (FBN2). PMID- 8401519 TI - Polymorphic dinucleotide repeats at the D3S1417, D3S1418 and D12S271 loci. PMID- 8401520 TI - Three polymorphic dinucleotide repeats near the von Hippel Lindau (VHL) disease gene on human chromosome 3: D3S587; D3S1317; D3S1435. PMID- 8401521 TI - Three tetranucleotide polymorphisms for loci: D3S1352; D3S1358; D3S1359. PMID- 8401522 TI - A chromosome 10p11.2 Gt-dinucleotide repeat polymorphism at the GLUDP5 gene locus. PMID- 8401523 TI - Trinucleotide repeat polymorphism at the D5S556 locus. PMID- 8401524 TI - Dinucleotide repeat polymorphism in the 3' untranslated region of an anonymous brain cDNA mapping to chromosome 2 (D2S230). PMID- 8401525 TI - Dinucleotide repeat polymorphism at the D10S469 locus. PMID- 8401526 TI - A triplet repeat polymorphism in a gene expressed in human hypothalamus. PMID- 8401527 TI - Dinucleotide repeat polymorphism at the D11S995 locus. PMID- 8401528 TI - SSCP polymorphism in exon 8B of the human G protein alpha O2 subunit (GNA01) gene. PMID- 8401529 TI - New human DNA polymorphisms submitted to the genome data base. PMID- 8401530 TI - A 2 base pair deletion in the RDS gene associated with butterfly-shaped pigment dystrophy of the fovea. PMID- 8401531 TI - Alternative splicing in the fragile X gene FMR1. PMID- 8401532 TI - A suggested nomenclature for designating mutations. PMID- 8401533 TI - Rhodopsin mutations in autosomal dominant retinitis pigmentosa. AB - Retinitis pigmentosa is an inherited progressive disease which is a major cause of blindness in western communities. It can be inherited as an autosomal dominant, autosomal recessive, or X-linked recessive disorder. In the autosomal dominant form (adRP), which comprises about 25% of total cases, approximately 30% of families have mutations in the gene encoding the rod photoreceptor-specific protein rhodopsin. This is the transmembrane protein which, when photoexcited, initiates the visual transduction cascade. So far, 41 single-base-pair (bp) substitutions, one two-bp substitution, and four deletions ranging from 3 to 42 bp have been identified in this gene. These mutations do not appear to be significantly clustered in a specific part of the protein, but occur in all three major domains, namely the intradiscal, transmembrane, and cytoplasmic regions. Different mutations appear to cause differences in the severity of the disease, though there is considerable variability in severity even within the same family, at least in certain of these mutations. Identification of all the mutations involved in rhodopsin-RP should allow accurate and early detection of affected individuals, informed genetic counselling, as well as furthering our knowledge of the disease process involved. PMID- 8401534 TI - A novel deletion in the low-density lipoprotein receptor gene in a patient with familial hypercholesterolemia from Petersburg. AB - Fifty St. Petersburg patients with type IIa hyperlipoproteinemia were screened for the presence of structural rearrangements in the low-density lipoprotein receptor (LDLR) gene. One novel deletion of the length about 5 kilobases (kb) was found. This deletion seems to remove completely exons 4, 5, and 6 from the LDLR gene, coding for the largest part of the receptor ligand-binding domain. PMID- 8401535 TI - Two mutations affecting the transport and maturation of lysosomal alpha glucosidase in an adult case of glycogen storage disease type II. AB - The autosomal recessive glycogen storage disease type II is associated with a deficiency of lysosomal alpha-glucosidase (acid maltase). This paper reports on the mutations in the lysosomal alpha-glucosidase alleles of an adult patient. A G 1927 to A transition was discovered in exon 14 causing the substitution of Gly 643 by Arg and a second C-2173 to T transition in exon 15 resulting in the substitution of Arg-725 by Trp. Each of the mutations was located in a different allele. The mutations were introduced in the wild-type lysosomal alpha glucosidase cDNA and expressed in COS cells. Both mutations had a similar effect. The synthesis of the mutant enzyme precursors was not disturbed but the intracellular transport and maturation were impaired. As a result there was an overall deficiency of catalytic activity. PMID- 8401536 TI - Screening for TP53 mutations in osteosarcomas using constant denaturant gel electrophoresis (CDGE). AB - We have previously developed conditions to screen for TP53 point mutations inside the conserved domains II-V of the gene by using constant denaturant gel electrophoresis (CDGE). The present study reports conditions for screening more of the codons in the frequently mutated region exon 5-8 and for detecting mutations in sequences encoding functional domains in the N- and C-terminal part of the protein. The ability of the CDGE technique to detect mutations was studied using controls with known sequence deviations. The resolution power of the technique to separate different types of mutations was tested by using seven different single base pair mutants all residing in a stretch of four base pairs. All mutants were separated from the wild type. The established CDGE screening strategy was then used to look for mutations in DNA from 28 osteosarcomas. Six (21.5%) of the samples were shown to have a TP53 mutation, and the exact characterization was performed by direct sequencing. All of these were within the frequently reported mutated region exon 5-8. PMID- 8401537 TI - Infidelity in the structure of ectopic transcripts: a novel exon in lymphocyte dystrophin transcripts. AB - Ectopic (or "illegitimate") transcripts have recently become popular as a means of facilitating the study of transcripts normally considered to have a pattern of expression restricted to one or a few tissues. It has been generally assumed that the structure of an ectopic transcript faithfully represents that of its tissue specific counterpart. We describe here the inclusion of a novel exon in 50% of ectopic dystrophin transcripts from human peripheral blood lymphocytes. The novel sequence resembles a conserved region in the 3' untranslated region of members of the carcinoembryonic antigen gene family and lies within the first intron of the human dystrophin gene. This constitutes a significant departure from the expected in vivo splicing behaviour in an ectopic transcript and suggests that there may be exceptions to the assumption that ectopic transcripts are processed in a similar way to their tissue-specific counterparts. PMID- 8401538 TI - Screening for the two most frequent mutations in Leber's hereditary optic neuropathy by duplex PCR based on allele-specific amplification. AB - This report describes a rapid and inexpensive assay, which allows detection, in whole blood and by PCR alone, of the two most frequent mitochondrial DNA mutations causing Leber's hereditary optic neuropathy. The assay is based on allele-specific amplification, using primers with the mutation-specific base in the 3' position, and a deliberately introduced G-->C substitution of base no. four from the 3' end, which prevents amplification of the wild-type allele. PMID- 8401539 TI - A novel nonsense mutation in the human dystrophin gene. PMID- 8401540 TI - Type A Niemann-Pick disease: a frameshift mutation in the acid sphingomyelinase gene (fsP330) occurs in Ashkenazi Jewish patients. PMID- 8401541 TI - Two new mutations at the adenosine deaminase (ADA) locus (Q254X and del nt1050 54) unusual for not being missense mutations. PMID- 8401542 TI - Screening for mutations in the antithrombin III gene causing recurrent venous thrombosis by single-strand conformation polymorphism analysis. PMID- 8401543 TI - A new delta-chain variant hemoglobin A2-Puglia or alpha 2 delta 2 26 Glu-->Asp (B8), detected by DNA analysis in a family of southern Italian origin. PMID- 8401544 TI - No C1840 to T mutation in RYR1 in malignant hyperthermia. PMID- 8401545 TI - Integrating the twelve-step approach with traditional psychotherapy for the treatment of eating disorders. AB - The treatment outcome literature for anorexia nervosa and bulimia indicates that approximately two thirds of patients treated with brief to moderate length interventions have favorable responses. The remaining one third are more difficult to treat and appear to require longer term and more intensive intervention. These patients can usually be characterized as having more severe Axis I and Axis II comorbidity. This manuscript focuses on a longer term treatment program that integrates psychodynamically oriented psychotherapy with a 12-step component. Specific issues that are addressed include the theoretical dilemma between the two models, advantages and disadvantages of adding a 12-step component, and how various subgroups of patients respond to the model. The clinical viability and preliminary impressions regarding outcome with this model are also offered. PMID- 8401546 TI - Outcome and prognostic variables in bulimia nervosa. AB - The purpose of this paper is to examine the factors that influence treatment response and outcome in 39 patients with bulimia nervosa who were assessed during the course of 8 weeks of cognitive-behavioral therapy, and after an 8-week and 1 year follow-up period. The patients' progress was assessed using data gathered from clinical examination, structured interviews, and self-rating scales. Patients who had a poor clinical response at the end of treatment had greater pretreatment symptom severity, lower body mass index, and were more likely to have personality disorders. Poor response after 1 year was associated with personality disorder, pretreatment symptom severity, and longer duration of illness. Patients without these poor prognostic indicators are more likely to respond to brief psychoeducational interventions. Patients with poor prognostic indicators are more suited to intensive psychological, pharmacologic, and experimental treatment approaches. PMID- 8401547 TI - Eating disorders among women receiving treatment for an alcohol problem. AB - To evaluate the frequency of eating disorder features among women receiving treatment for an alcohol problem, 52 female attenders at an alcohol clinic were assessed using a standardized research clinical interview, and the findings compared with data on a general population sample. The results suggest that eating disorder features are overrepresented among women who present for treatment for an alcohol problem. PMID- 8401548 TI - The relation among stress, psychological symptoms, and eating disorder symptoms: a prospective analysis. AB - Prior research has shown that psychopathology and stress are associated with eating disorder symptoms, but the relations among these variables were confounded by concurrent measurement at a single point in time. The present study examined the relations among stress, psychological, and eating disorder symptoms prospectively over 4 months in 143 adolescent girls. In cross-sectional analyses at follow-up, controlling for baseline levels, stress and eating disorder symptoms each predicted changes in the other. Eating disorder symptoms were not predicted by psychological symptoms nor vice versa. Stress was predicted by eating disorder symptoms from the first assessment to follow-up. On balance, the results are somewhat stronger for the argument that eating disorder symptoms are predictive of subsequent psychological stress over short periods of time. PMID- 8401549 TI - Depression in anorexia nervosa at follow-up. AB - This study set out to investigate the correlation between depressive symptomatology and the eating disorder psychopathology at a 3-year follow-up after discharge of 34 adolescent anorexia nervosa patients. Three standardized rating instruments were employed to assess depression and two defined outcome criteria were used to assess the eating disorder. Correlation analysis revealed that there was a consistent and significant association between the severity of eating disorder symptoms and the degree of depression at follow-up. According to the subscales of the average outcome score of Morgan and Russell (Psychological Medicine 5:355-371, 1975), depression was related more to social maladaptation than to the specific eating disorder psychopathology. PMID- 8401550 TI - Effects of eating behavior on mood: a review of the literature. AB - The literature focusing on the use of food as a regulator of a negative mood state is reviewed. This literature reveals that individuals experiencing a negative mood state arising from disorders ranging from tobacco withdrawal to premenstrual symptoms make use of carbohydrate ingestion, especially simple carbohydrates, to provide a temporary lifting of mood. However, other evidence suggests that some individuals may obtain a more permanent control of their negative mood stay by eliminating simple carbohydrates from their diet. While the literature is consistent in demonstrating that carbohydrate consumption can alter a negative mood state, the underlying mechanism mediating this relationship is unknown. PMID- 8401551 TI - Cognitive changes to preloading in restrained and unrestrained eaters as measured by the Stroop task. AB - Restraint theory has identified overeating in response to a high calorie preload as characteristic of restrained eaters. The present study evaluated cognitive and motivational changes to preloading using both self-report rating scales and Stroop tasks. The results suggest that the restrained eaters responded to a high calorie preload with increased feelings of rebelliousness, defiance, and a desire to challenge the limitations set by the diet, described as an active state of mind, as measured by both the Stroop task and the rating scales. The restrained eaters also showed retardation in the color naming of the body size words and food words after the high calorie preload, suggesting that the consumption of a forbidden food may increase the dieter's concern about food and her shape and weight. The results are discussed in terms of the effects of preloading and possible mediating variables involved in the transition from successful restraint to overeating. PMID- 8401552 TI - Idiopathic edema and eating disorders: evidence for an association. AB - Using a computerized self-report questionnaire format incorporating the Eating Attitudes Test-26 (EAT-26) and a modification of the McKendry criteria for the diagnosis of idiopathic edema (IE), the authors surveyed a population of women students to determine the prevalence of IE symptoms and their relationship to abnormal eating attitudes and behaviors (AEAB). The mean age of 177 respondents was 20.8 years. IE symptoms were reported by 12.4% of respondents, and AEAB by 16.4%. The prevalence of IE symptoms was 37.9% in respondents with AEAB and 7.4% in normal respondents (p = .0001). A score of 11 or greater on the modified McKendry IE criteria had a sensitivity of .37, specificity of .93, positive predictive value of .5, and a negative predictive value of .89 for an abnormal EAT-26 score. Body mass index (BMI) scores were similar for all respondent groups. PMID- 8401553 TI - Eating, weight, and dieting disturbances in male and female lightweight and heavyweight rowers. AB - Rowers compete in a sport that allows comparison of male and female athletes and where some (lightweight) but not others (heavyweights) must meet specific weight criteria. Eating attitudes, dieting patterns, weight fluctuation, and methods of weight loss were evaluated in 162 rowers: 82 heavyweights (56 females, 26 males) and 80 lightweights (17 females, 63 males). Females displayed more disturbed eating practices and weight control methods than did males. Lightweights did not have more disturbed eating practices than heavyweights, but employed more extreme weight loss methods. Male rowers were more affected by weight restriction than were female rowers. Lightweight males showed greater weight fluctuation during the season and gained more weight during the offseason than did lightweight females and heavyweight males and females. These results indicate that rowing can join the growing list of sports where eating and weight disturbances may be present. Male athletes may be more vulnerable to these problems than previously recognized. PMID- 8401554 TI - The Tridimensional Personality Questionnaire in eating disorder patients. AB - The Tridimensional Personality Questionnaire (TPQ) was developed to measure a variety of personality variants on three biosocial dimensions, harm avoidance (HA), novelty seeking (NS), and reward dependence (RD), which are thought to be related to serotonin (5-HT), dopamine (DA), and norepinephrine (NE) function, respectively. Patients with eating disorders have been reported to have abnormalities in all of these systems, as well as personality variants described by these dimensions. We therefore administered the TPQ to 147 patients with DSM III-R defined eating disorders (110 bulimia nervosa [BN], 27 with anorexia nervosa [AN], and 10 with BN+AN) and compared their scores to those of 350 female controls. When significant, post hoc Bonferroni t tests were performed using alpha = 0.05. All subtypes of eating disorder patients scored significantly higher on HA than controls (p < or = .0001, analysis of variance. Only patients with BN (+/- AN) had significantly higher degrees of NS (p < or = .0001), particularly on the impulsiveness subscale (NS2), although this may, in part, be due to age. No significant differences in total RD were found, although BN patients scored lower on RD3 (attachment vs. detachment) and higher on RD4 (dependence vs. independence) than controls. In addition, AN patients had significantly higher RD2 (persistence vs. irresoluteness) subscale scores. These data support a theory of 5-HT dysregulation in both types of eating disorders and suggest that further research be done on the role of DA and NE in BN. PMID- 8401555 TI - Alexithymia in the eating disorders. AB - Eating disorder patients appear to have high degrees of alexithymia, a diminished capability to verbally describe feelings, although little data exist. We administered the Toronto Alexithymia Scale (TAS) to 114 females with DSM-III-R defined eating disorders. Patients, regardless of subtype, scored significantly higher than 370 college-aged females. TAS scores were significantly correlated to self-ratings of affective symptoms, but not weight or binge-purge frequency. PMID- 8401556 TI - Depressed mood and concern with weight and shape in normal young women. AB - The possible role of depression in accentuating the concerns with weight and shape found in some members of the normal female population was investigated using mood induction procedures. A depressed mood was induced in two groups of normal young women; a group placing a high personal value on shape or weight, and a group placing a low value on shape and weight. The concerns with shape were significantly elevated in the former group compared with the latter. This result is consistent with a mechanism for the development of a disturbance in body image proposed by Cooper and Taylor (British Journal of Psychiatry 153 [Suppl. 2] 20 22, 1988). PMID- 8401557 TI - Denervation of the rat posterior pituitary gland: validation of a stereotaxic method. AB - A stereotaxic surgical method was developed for interrupting the nerve fibres running through the rat pituitary stalk to the posterior pituitary gland without obliterating the hypothalamo-pituitary portal circulation. The pituitary stalk was compressed by the blunt tip of an L-shaped rotating knife. Successful operations produced mild diabetes insipidus, disappearance of arginine vasopressin from the neural lobe, accumulation of arginine vasopressin and neurosecretory material in the pituitary stalk and no infarction in the anterior lobe of the pituitary gland. In female rats, the oestrous cycle was only temporarily disturbed. Plasma prolactin and corticosterone levels were high during the first 24 h after the stalk compression but returned to normal baseline levels from the second day after the operation. One week after the operation plasma adrenocorticotropin and prolactin levels were in the control range while plasma alpha-melanocyte-stimulating hormone was elevated. Denervation of the posterior pituitary gland may help in studying the neural control of intermediate lobe function and the role of the neural lobe in various endocrine conditions, and may serve as a model for lesions of the pituitary stalk and formation of ectopic neurohypophysis in the human. PMID- 8401558 TI - Local tetrodotoxin blocks chronic stress effects on corticotropin-releasing factor and vasopressin messenger ribonucleic acids in hypophysiotropic neurons. AB - To test the hypothesis that synaptic inputs to the paraventricular nucleus mediate stress-induced increases in corticotropin-releasing peptide expression in the paraventricular nucleus of the hypothalamus (PVH), relative levels of the mRNAs encoding corticotropin-releasing factor (CRF) and arginine vasopressin (AVP) were followed, in situ, in animals subjected to chronic footshock stress and concurrent local administration of tetrodotoxin or vehicle. Consistent with previous findings, a 7-day exposure to chronic footshock resulted in a 2.1-fold increase in CRF mRNA levels in the parvocellular division of the PVH. The footshock paradigm also resulted in at least a 41% increase in AVP transcripts in this same region; this effect was localized predominantly to CRF-immunoreactive neurons. The stressor did not significantly alter AVP mRNA levels in the magnocellular division of the PVH. Tetrodotoxin, administered to the PVH via osmotic minipump, blocked the stress-induced rise in CRF and AVP mRNAs, but had no significant effect on basal levels of these transcripts. The results support the view that maintenance of the enhanced central drive on pituitary-adrenal activity seen in response to chronic stress is mediated via neural inputs to the PVH. PMID- 8401559 TI - Androgenic regulation of expression of beta-tubulin messenger ribonucleic acid in motoneurons of the spinal nucleus of the bulbocavernosus. AB - Expression of beta-tubulin mRNA was examined in androgen-sensitive motoneurons of the spinal nucleus of the bulbocavernosus (SNB) in adult male rats by in situ hybridization histochemistry using cDNA encoding mouse beta-tubulin. Hybridizable beta-tubulin mRNA was localized in the somata and proximal dendrites of SNB motoneurons. Removal of androgen by castration significantly reduced the expression level of beta-tubulin mRNA in the SNB motoneurons, whereas the change was prevented by testosterone treatment. On the contrary, castration or testosterone treatment did not induce any changes in the expression level of beta tubulin mRNA in the androgen-insensitive motoneurons of the retrodorsolateral nucleus. These results suggest that androgen regulates the expression of beta tubulin gene in the SNB motoneurons and may provide evidence for the molecular mechanisms of hormonally-induced neuronal plasticity in the SNB motoneurons. PMID- 8401560 TI - Effects of novelty stress on vasopressin and oxytocin secretion by the pituitary in the rat. AB - Effects of novel environmental stimuli on vasopressin and oxytocin secretion by the pituitary were studied in dehydrated male rats. As the novel environmental stimuli, rats were transferred to an experimental room, placed in a box painted black and given a pure tone auditory stimulus of 2 kHz. Exposure of rats to the novel environmental stimuli for a period of 2 min decreased plasma concentrations of vasopressin and increased plasma levels of adrenocorticotrophic hormone (ACTH) and prolactin, but did not significantly change the plasma level of oxytocin. The stimuli, however, became ineffective for producing the suppressive vasopressin response as the period of exposure was prolonged to more than 5 and up to 30 min, although the prolonged stimuli were still effective for inducing facilitatory ACTH and prolactin responses. After repeated exposures of rats to the environmental stimuli once a day for 5 or 10 days, the stimuli became disabled from producing the suppressive vasopressin response. However, the rats were still capable of responding to the novel stimuli of another kind. All these data suggest that novelty stress suppresses vasopressin secretion but does not change oxytocin secretion. In order to test the possibility that glucocorticoids expectedly secreted by the adrenals in response to the stress might have suppressed vasopressin secretion, a large amount of dexamethasone was administered to the rat before testing. Dexamethasone pretreatment depressed plasma levels of ACTH and vasopressin as reported previously and blocked the facilitatory ACTH response to the novelty stress. However, dexamethasone treatment did not affect the suppressive vasopressin response to the novelty stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401561 TI - Hypothalamic neuropeptide Y messenger ribonucleic acid levels in pre-obese and genetically obese (fa/fa) rats; potential regulation thereof by corticotropin releasing factor. AB - Neuropeptide Y (NPY) is a 36 amino-acid peptide. It is localized within the brain but is also present peripherally. It is a well substantiated orexigenic peptide with several other endocrine and behavioural effects. In this study NPY mRNA levels were measured, using the polymerase chain reaction amplification technique, in the hypothalamus of pre-obese (unweaned 13-day-old), young (weaned 28-day-old) and adult (11-week-old) obese fa/fa rats and compared to those of lean age-matched controls. Before weaning, pre-obese pups had the same NPY mRNA levels as controls. After weaning NPY mRNA levels were increased 2-fold in young 28-day-old and 4-fold in adult obese rats, relative to corresponding controls. When adult obese rats were intracerebroventricularly-treated with ovine corticotropin-releasing hormone (oCRF) for 7 days, they stopped gaining body weight relative to vehicle-infused obese controls. Upon measuring NPY mRNA levels in the hypothalamus of these two groups of animals, it was shown that the high NPY mRNA levels of vehicle-treated (control) obese rats were decreased by 3-fold following the intracerebroventricular oCRF administration. It is proposed that: 1) hypothalamic NPY may play a role in the establishment and maintenance of the genetic obesity syndrome of the fa/fa rat, and 2) maintenance of the genetic obesity syndrome of the fa/fa rat, and 2) hypothalamic NPY could be partly regulated by central CRF. PMID- 8401562 TI - Ultrastructural evidence for changes in synaptic input to the hypothalamic luteinizing hormone-releasing hormone neurons in photosensitive and photorefractory starlings. AB - Neural input to the hypothalamic luteinizing hormone-releasing hormone (LHRH) neurons was investigated in male starlings (Sturnus vulgaris) using electron microscopy combined with immunocytochemistry. Birds (4 to 6 in each group) were sampled at four stages of a photoperiodically induced reproductive cycle: (a) sexually immature but photosensitive, under short days; (b) during sexual maturation after 7 to 25 long days; (c) during gonadal regression after 35 to 50 long days; and (d) when fully photorefractory after 11 months exposure to long days. The length of the perikaryal membrane, the number and length of axo-somatic terminals in contact with it and the number and length of synaptic modifications within the terminals were measured for a minimum of six LHRH neurons in each brain. The number of axo-somatic terminals per neuron and the number per unit length of perikaryal membrane did not differ in birds of groups (a), (b) and (c), but was significantly greater (P < 0.05) in the fully refractory birds (group d). Similarly, the number of synaptic modifications was significantly greater (P < 0.05) in group (d) than in the other groups. These results are consistent with increased neural input to the LHRH perikarya in photorefractory birds after prolonged exposure to long days, although there was no indication of a change in input at the time of gonadal regression. PMID- 8401563 TI - Fos expression in the rat brain following vaginal-cervical stimulation by mating and manual probing. AB - Vaginal-cervical stimulation (VCS), provided by mating or manual probing, induces many reproductive behavioral and endocrine changes in female rats. These changes include an increase in lordosis duration, heat termination and pseudopregnancy. Electrophysiological and [14C]2-deoxy-D-glucose studies collectively show that neurons in the medial preoptic area, ventromedial hypothalamus and midbrain central gray respond to manual VCS. In the present study we immunocytochemically labeled brain sections for Fos, the protein product of the immediate early gene c fos, to detect VCS-responsive neurons in hormone-primed animals receiving VCS by mating or manual probing. In Experiment 1, females receiving mounts and intromissions were compared to: 1) vaginally-masked females receiving mounts but no VCS, 2) females exposed to an intact anesthetized male or 3) females not exposed to males or the testing arena. Those animals receiving VCS showed a dramatic increase in the number of Fos-immunoreactive cells in the medial preoptic area, posterodorsal portion of the medial amygdala and bed nucleus of the stria terminalis, as well as the dorsomedial hypothalamus, ventromedial hypothalamus and midbrain central gray. These effects of VCS were confirmed in Experiment 2 in animals receiving manual vaginal-cervical probing. These findings extend previous electrophysiological and [14C]2-deoxy-D-glucose studies by providing evidence that additional brain areas respond to VCS by mating, as well as manual probing. PMID- 8401564 TI - Vasostatins, comprising the N-terminal domain of chromogranin A, suppress tension in isolated human blood vessel segments. AB - Chromogranin A (CGA) belongs to a family of highly acidic proteins which are co stored and co-released with the catecholamines from the mammalian adrenal gland and occur in nmolar concentrations in the human circulation. A vascular function for the adrenomedullary released and circulating CGA has yet to be established. The present study reports on the novel vasoinhibitory effect of the N-terminal domain of the adrenomedullary CGA in isolated segments of the human internal thoracic artery (ITA) and saphenous vein (SV). The collective term vasostatin(s) refers to N-terminal fragments (CGA1-76 and CGA1-113) of apparent molecular weights 7 to 22 kD, to indicate their vascular inhibitory effects. The sustained contractions evoked by the potent vasoconstrictor peptide, endothelin-1 (ET-1) were suppressed when ITA and SV segments were preincubated for 15 min with vasostatins (72 nM). The vasoinhibitory effects were not dependent on an intact endothelium and suppression of the response to 35 nM ET-1 was approximately 77% and approximately 40% in endothelium-denuded ITA and SV segments, respectively. In endothelium-denuded SV segments the vasostatins suppressed the maximal sustained tension response but not the potency for ET-1, indicating that the vasostatin effect did not involve interference with ET-1 binding to its vascular receptor. Preincubation of endothelium-denuded SV segments with nifedipine (1 microM) inhibited the sustained response to ET-1 > or = 10 nM by 50%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401565 TI - Chronic but not acute exposure to stress is associated with hypothalamic vasoactive intestinal polypeptide (VIP) release into median eminence. AB - The influence of stress on hypothalamic VIP release into the pituitary portal blood has not been assessed at present despite the fact that this peptide has been implicated in the control of several pituitary hormones and especially in the release of prolactin (PRL) caused by stress. In the present work the effect of stress on the in vivo release of VIP into the pituitary portal blood of male rats was evaluated by means of push-pull perfusion (PPP) of median eminence (EM). VIP content in the PPP liquid was successfully measured and their levels agree well with pituitary portal blood levels measured by other authors. Whereas plasma PRL levels strongly increased during acute immobilization (IMO), no changes in VIP secretion into the ME were observed. VIP release into the ME was also unaffected by exposure to ether. In contrast, in chronically immobilized rats a significant increase in VIP release into the ME was obtained in response to acute IMO. The present data argue against a major role of hypothalamic VIP in PRL release caused by acute stress and show that chronic stress qualitatively alters the signal of hypothalamic VIP to the pituitary. PMID- 8401566 TI - The actions of endothelin on single cells in the anteroventral third ventricular region and supraoptic nucleus in rat hypothalamic slices. AB - Endothelin (ET), a peptide consisting of 21 amino-acid residues was recently isolated from the culture supernatant of porcine aortic endothelial cells. ET has been reported to be a more potent vasoconstrictor than angiotensin II. Other studies suggest that ET is involved in central control of the autonomic nervous system and body water regulation. Extracellular recordings were made from neurons in the anteroventral third ventricle (AV3V) and supraoptic nucleus (SON) in rat hypothalamic slice preparations. ET-3 was applied at concentrations of 10(-10) M to 3 x 10(-7) M. Of 226 AV3V neurons tested, 48 (21%) were excited, 8 (4%) were inhibited, and 170 (75%) were unaffected by ET-3 at 10(-7) M. The threshold concentration to evoke the responses was approximately 10(-9) M. Of 144 SON neurons tested, 64 had a phasic firing pattern and 80 had a non-phasic firing pattern. Of 64 phasic neurons tested, 39 (61%) were inhibited by ET-3 at 10(-7) M, 25 (39%) were non-responsive and none was excited. Of 80 non-phasic neurons tested, 14 (17.5%) were inhibited by ET-3 at 10(-7) M, 66 (82.5%) were non responsive and none was excited. The effects of ET-1 were compared with those of ET-3. The number of neurons responding to ET-1 and their responsiveness were almost the same as for ET-3. To investigate whether the ET responses are dependent on Ca2+ influx, a Ca2+ free medium and the Ca2+ antagonist, nicardipine, were used. The excitatory responses of AV3V neurons to ET were maintained in the Ca2+ free medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401567 TI - Changes in oxytocin immunoreactivity and mRNA expression in the sheep brain during pregnancy, parturition and lactation and in response to oestrogen and progesterone. AB - The effects of pregnancy, parturition and lactation and exogenous treatments with oestradiol and progesterone on oxytocin (OXY) immunoreactivity and gene expression in the sheep brain were investigated. Immunocytochemistry was used to demonstrate that increased OXY-immunoreactivity occurred in cells of the paraventricular (PVN) and supraoptic nuclei (SON), the bed nucleus of the stria terminalis (BNST), the anterior commissural nuclei (ACN) and the periventricular part of the medial preoptic area (PvMP). Oxytocin immunoreactive terminals were also seen in the accessory olfactory nucleus, the glomerular and peri-glomerular layers of the olfactory bulb, the lateral septum, the zona incerta and the pars compacta of the substantia nigra. Compared to ovariectomized and late pregnant animals, the intensity of immunoreactivity was increased in all of these oxytocinergic elements at parturition, during lactation and following exogenous treatment with oestradiol. The OXY-immunoreactivity was also more intense in late pregnant animals compared to ovariectomized ones. Quantitative in situ hybridization histochemistry showed that cells in the PVN, SON, BNST and PvMP all showed significantly increased expression of OXY mRNA in animals at parturition and during lactation compared to late pregnant or ovariectomized animals. Expression levels in late pregnant animals were also significantly higher than in ovariectomized ones. Progesterone treatment significantly increased OXY mRNA in the PVN, SON, BNST and PvMP whereas oestradiol treatment was only effective in the PVN, BNST and PvMP. Combined treatment with these steroids did not significantly increase OXY mRNA levels in comparison with their administration alone. These results show that OXY-immunoreactivity and mRNA expression are at their highest in the sheep brain when maternal behaviour is induced. The increased synthesis/storage of the peptide at parturition may be due to changes in circulating concentrations of both progesterone and oestradiol during late pregnancy. PMID- 8401568 TI - Centrally injected interleukin-1 beta inhibits the hypothalamic LHRH secretion and circulating LH levels via prostaglandins in rats. AB - Intracerebroventricular (icv) infusion of interleukin-1 beta (IL-1 beta) significantly lowers plasma LH levels in castrated male rats, and interferes with LHRH release into the median eminence of proestrus female rats. We have investigated the potential role of arachidonic acid metabolites in mediating these inhibitory effects, by administering indomethacin (INDO, a cyclooxygenase inhibitor) or nor- dihydroguaiaretic acid (NDGA, a lipoxygenase inhibitor) 15 min prior to injection of the cytokine. While not measurably altering basal LH or LHRH secretion in castrated or proestrus rats, respectively, INDO completely reversed the action of IL-1 beta on the secretion of these 2 hormones. In contrast, NDGA did not alter IL-1-induced decreases in LH release. The peripheral administration of endotoxin (LPS) also interferes with LH release. Because the iv injection of IL-1 does not alter LH secretion, this effect is believed to be at least in part mediated by increased synthesis of cytokines within the CNS. We observed that in contrast to results obtained in rats injected with IL-1 icv, INDO did not reverse the inhibitory action of LPS. Our results thus suggest either that central IL-1 is not the primary modulator of LPS-induced decrease in LH values, or that pathways other than those involving arachidonic acid are important.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401569 TI - Involvement of central corticotropin-releasing factor (CRF) in suckling-induced inhibition of luteinizing hormone secretion in lactating rats. AB - The lack of ovulation and the inhibition of reproductive functions observed in many species during lactation is closely related to the intensity of the suckling stimulus. However, the mechanisms by which suckling inhibits hypothalamic GnRH and pituitary LH secretion in rats are still unclear. Since we recently demonstrated that suckling is a persistent stimulus to the adrenococortical system of the rat, we tested the hypothesis that suckling-induced activation of central CRF release may mediate the associated inhibition of GnRH secretion. Lactating females were ovariectomized (OVX) on day 2 of lactation, and equipped with icv guide cannula on day 2 and indwelling jugular catheters on day 5 before testing on day 7. Lactating females were separated from their pups for 24 h prior to the suckling test with the following pretreatments: 1) icv injection with artificial CSF (aCSF) or a specific CRF antagonist, alpha-helical CRF (9-41), (25 micrograms/rat, CRF-AX) 15 min prior to pup reunion or 2) iv injection of normal sheep serum (NSS) or CRF antiserum (CRF-AB) 4 h prior to pup reunion. Plasma ACTH, LH and PRL concentrations were determined prior to and at various intervals after pup reunion. After 3 h of suckling, LH and PRL responses to a bolus injection of GnRH (10 ng/rat) were measured; a bolus injection of Angiotensin II (AII, 5 micrograms/rat) was administered after 4 h to test for ACTH responses. Non-lactating females injected with GnRH and AII were used as controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401570 TI - A functional promoter directing expression of a novel type of rat mineralocorticoid receptor mRNA in brain. AB - The authors have identified two types of hippocampal cDNAs for the rat mineralocorticoid receptor (rMR) which are identical in the protein coding domain but differ in their 5'-untranslated sequences. One of these clones encodes a novel type of rMR cDNA with a high homology to a previously described human MR cDNA isolated from the kidney. A genomic clone containing the 5'-end of the rat MR gene was isolated. The 12.7 kb genomic region contains the 5'-coding exon with the translational start site and contiguous DNA sequences encoding the N-terminal domain of the rMR. A 240 bp region homologous to the 5'-untranslated sequences of the novel rMR cDNA was located 5.2 kb upstream the protein coding region. Characterization of the nucleotide sequence preceding this exon revealed several features characteristic for promoters of so-called 'housekeeping genes'. The sequence analyzed is 635 bp in length, is rich in G+C nucleotides (63%) and lacks TATA or CAAT regulatory elements. It contains three putative binding sites for transcription factor Sp1 as well as several short sequences that are similar to known cis-acting enhancers or binding sites for transcription factors. In transient transfection experiments employing the luciferase reporter gene and the CV1 cell line this region exhibits substantial promoter activity. These experiments demonstrate that expression of the rat MR gene in the hippocampus results in at least two transcripts with different 5'-untranslated exons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401571 TI - Collaboration, Canadian style. PMID- 8401572 TI - Mapping the way ahead. PMID- 8401573 TI - Wither polygenic inheritance: mapping Hirschsprung disease. PMID- 8401574 TI - Livestock genome research on the march. PMID- 8401575 TI - Huntington's disease: testing the test. PMID- 8401576 TI - A missense mutation confirms the L1 defect in X-linked hydrocephalus (HSAS) PMID- 8401577 TI - dbEST--database for "expressed sequence tags". PMID- 8401578 TI - The FMR-1 protein is cytoplasmic, most abundant in neurons and appears normal in carriers of a fragile X premutation. AB - Fragile X mental retardation syndrome is caused by the unstable expansion of a CGG repeat in the FMR-1 gene. In patients with a full mutation, abnormal methylation results in suppression of FMR-1 transcription. FMR-1 is expressed in many tissues but its function is unknown. We have raised monoclonal antibodies specific for the FMR-1 protein. They detect 4-5 protein bands which appear identical in cells of normal males and of males carrying a premutation, but are absent in affected males with a full mutation. Immunohistochemistry shows a cytoplasmic localization of FMR-1. The highest levels were observed in neurons, while glial cells contain very low levels. In epithelial tissues, levels of FMR-1 were higher in dividing layers. In adult testis, FMR-1 was detected only in spermatogonia. FMR-1 was not detected in dermis and cardiac muscle except under pathological conditions. PMID- 8401579 TI - EWS and ATF-1 gene fusion induced by t(12;22) translocation in malignant melanoma of soft parts. AB - The genes involved in the t(12;22)(q13;q12) translocation found recurrently in malignant melanoma of soft parts have been characterized and shown to form, in four cases studied, hybrid transcripts. The deduced chimaeric protein encoded by the der(22) chromosome consists of the N-terminal domain of EWS linked to the bZIP domain of ATF-1, a transcription factor which may normally be regulated by cAMP. ATF-1 has not previously been implicated in oncogenesis. EWS was first identified as forming a hybrid transcript in Ewing's sarcoma, which links its N terminal domain to the DNA binding domain of the FLI-1 gene. Thus the oncogenic conversion of EWS follows a common scheme of activation, exchanging its putative RNA binding domain with different DNA binding domains that appear to be tumour specific. PMID- 8401580 TI - A gene for Hirschsprung disease maps to the proximal long arm of chromosome 10. AB - Hirschsprung disease (HSCR) is a frequent congenital disorder (1 in 5,000 newborns) of unknown origin characterized by the absence of parasympathetic intrinsic ganglion cells of the hindgut. Taking advantage of a proximal deletion of chromosome 10q (del 10q11.2-q21.2) in a patient with total colonic aganglionosis, and of a high-density genetic map of microsatellite DNA markers, we performed genetic linkage analysis in 15 non-syndromic long-segment and short segment HSCR families. Multipoint linkage analysis indicated that the most likely location for a HSCR locus is between loci D10S208 and D10S196, suggesting that a dominant gene for HSCR maps to 10q11.2, a region to which other neural crest defects have been mapped. PMID- 8401581 TI - A gene for Hirschsprung disease (megacolon) in the pericentromeric region of human chromosome 10. AB - Hirschsprung disease (HSCR) is characterized by a congenital absence of enteric ganglia along a variable length of the intestine. Although long considered to be a multifactorial disease, we have identified linkage in a subset of five HSCR families to the pericentromeric region of chromosome 10, thereby providing monogenic inheritance in some families. A maximum two-point lod score of 3.37 (theta = 0.045) was observed between HSCR and D10S176, under an incompletely penetrant dominant model. Multipoint, affecteds-only and non-parametric analyses supported this finding and localize this gene to a region of approximately 7 centiMorgans, in close proximity to the locus for multiple endocrine neoplasia type 2 (MEN2). The co-occurrence of these two entities in some families might be attributable to shared pathogenetic origins. PMID- 8401582 TI - A missense mutation in the dystrophin gene in a Duchenne muscular dystrophy patient. AB - About two thirds of Duchenne muscular dystrophy (DMD) patients have either gene deletions or duplications. The other DMD cases are most likely the result of point mutations that cannot be easily identified by current strategies. Utilizing a heteroduplex technique and direct sequencing of amplified products, we screened our nondeletion/duplication DMD population for point mutations. We now describe what we believe to be the first dystrophin missense mutation in a DMD patient. The mutation results in the substitution of an evolutionarily conserved leucine to arginine in the actin-binding domain. The patient makes a dystrophin protein which is properly localized and is present at a higher level than is observed in DMD patients. This suggests that an intact actin-binding domain is necessary for protein stability and essential for function. PMID- 8401583 TI - A NotI restriction map of the entire long arm of human chromosome 21. AB - A variety of maps of the human genome have been constructed, including cloned DNA maps. We have isolated 40 of the 42 NotI sites that exist on the long arm of human chromosome 21, as NotI linking clones and constructed a complete NotI restriction map spanning the entire region. This map, which provides the most reliable ordering and distance estimation in the region from a pericentromeric locus to the terminus, demonstrates the usefulness of linking clone mapping for analysing human chromosomes. PMID- 8401584 TI - Genomic scanning for expressed sequences in Xp21 identifies the glycerol kinase gene. AB - Rapid genomic scanning methods are required to identify expressed sequences and we report an efficient, sensitive and specific approach which relies upon hybridization of an amplified, labeled cDNA library to digested cosmid DNA. We identified expressed sequences within a cosmid in the glycerol kinase (GK) "critical region" of Xp21 that had impressive similarity to prokaryotic GKs. We used this genomic sequence information to clone the human hepatic GK cDNA. Independent confirmation of the identity of this gene was obtained by functional complementation of GK deficient E. coli mutants with a construct containing the complete human X-linked GK coding sequence. PMID- 8401585 TI - Rapid cDNA sequencing (expressed sequence tags) from a directionally cloned human infant brain cDNA library. AB - A human infant brain cDNA library, made specifically for production of expressed sequence tags (ESTs) was evaluated by partial sequencing of over 1,600 clones. Advantages of this library, constructed for EST sequencing, include the use of directional cloning, size selection, very low numbers of mitochondrial and ribosomal transcripts, short polyA tails, few non-recombinants and a broad representation of transcripts. 37% of the clones were identified, based on matches to over 320 different genes in the public databases. Of these, two proteins similar to the Alzheimer's disease amyloid precursor protein were identified. PMID- 8401586 TI - Chromosomal distribution of 320 genes from a brain cDNA library. AB - We have determined the chromosomal assignment of 320 brain expressed genes by studying the segregation of polymerase chain reaction (PCR) products in human rodent somatic cell hybrids and by genetically mapping polymorphic cDNAs using the CEPH (Centre d'Etude du Polymophisme Humaine) reference pedigrees and database. These mapped genes can function as markers on the physical map of the human genome, as well as serve as candidate disease gene loci. Distribution of these genes to the human chromosomes correlates well with the GC content of the chromosomes. However, the distribution of these genes does not correlate well with the cytogenetic length of each chromosome. PMID- 8401587 TI - Trinucleotide repeat length instability and age of onset in Huntington's disease. AB - The initial observation of an expanded and unstable trinucleotide repeat in the Huntington's disease gene has now been confirmed and extended in 150 independent Huntington's disease families. HD chromosomes contained 37-86 repeat units, whereas normal chromosomes displayed 11-34 repeats. The HD repeat length was inversely correlated with the age of onset of the disorder. The HD repeat was unstable in more than 80% of meiotic transmissions showing both increases and decreases in size with the largest increases occurring in paternal transmissions. The targeting of spermatogenesis as a particular source of repeat instability is reflected in the repeat distribution of HD sperm DNA. The analysis of the length and instability of individual repeats in members of these families has profound implications for presymptomatic diagnosis. PMID- 8401588 TI - Relationship between trinucleotide repeat expansion and phenotypic variation in Huntington's disease. AB - The molecular analysis of a specific CAG repeat sequence in the Huntington's disease gene in 440 Huntington's disease patients and 360 normal controls reveals a range of 30-70 repeats in affected individuals and 9-34 in normals. We find significant negative correlations between the number of repeats on the HD chromosome and age at onset, regardless of sex of the transmitting parent, and between the number of repeats on the normal paternal allele and age at onset in individuals with maternally transmitted disease. This effect of the normal paternal allele may account for the weaker age at onset correlation between affected sib pairs with disease of maternal as opposed to paternal origin and suggests that normal gene function varies because of the size of the repeat in the normal range and a sex-specific modifying effect. PMID- 8401589 TI - The relationship between trinucleotide (CAG) repeat length and clinical features of Huntington's disease. AB - Huntington's disease (HD) is associated with the expansion of a CAG trinucleotide repeat in a novel gene. We have assessed 360 HD individuals from 259 unrelated families and found a highly significant correlation (r = 0.70, p = 10(-7)) between the age of onset and the repeat length, which accounts for approximately 50% of the variation in the age of onset. Significant associations were also found between repeat length and age of death and onset of other clinical features. Sib pair and parent-child analysis revealed that the CAG repeat demonstrates only mild instability. Affected HD siblings had significant correlations for trinucleotide expansion (r = 0.66, p < 0.001) which was not apparent for affected parent-child pairs. PMID- 8401590 TI - Polygenic control of autoimmune diabetes in nonobese diabetic mice. AB - Partial exclusion mapping of the nonobese (NOD) diabetic mouse genome has shown linkage of diabetes to at least five different chromosomes. We have now excluded almost all of the genome for the presence of susceptibility genes with fully recessive effects and have obtained evidence of linkage of ten distinct loci to diabetes or the prediabetic lesion, insulitis, indicative of a polygenic mode of inheritance. The relative importance of these loci and their interactions have been assessed using a new application of multiple polychotomous regression methods. A candidate disease gene, interleukin-2 (Il-2), which is closely linked to insulitis and diabetes, is shown to have a different sequence in NOD, including an insertion and a deletion of tandem repeat sequences which encode amino acid repeats in the mature protein. PMID- 8401591 TI - The ovine Booroola fecundity gene (FecB) is linked to markers from a region of human chromosome 4q. AB - The autosomal Booroola fecundity gene (FecB) mutation in sheep increases ovulation rate and litter size, with associated effects on ovarian physiology and hormone profiles. Analysis of segregation in twelve families (379 female progeny) identified linkage between the mutation, two microsatellite markers (OarAE101 and OarHH55, Zmax > 9.0) and epidermal growth factor (EGF) from human chromosome 4q25 (Zmax > 3.0). The marker OarAE101 was linked to secreted phosphoprotein 1 (SPP1, which maps to chromosome 4q21-23 in man) in the test pedigrees and independent families (Zmax > 9.7). The identification of linkage between the FecB mutation and markers from human chromosome 4q is an important step towards further understanding the control of ovulation rates in mammals. PMID- 8401592 TI - The Wilms tumour gene WT1 is expressed in murine mesoderm-derived tissues and mutated in a human mesothelioma. AB - The tumour suppressor gene WT1 encodes a transcription factor expressed in tissues of the genito-urinary system. Inactivation of this gene is associated with the development of Wilms tumour a pediatric kidney cancer. We show that WT1 is also expressed at high levels in many supportive structures of mesodermal origin in the mouse. We also describe a case of adult human mesothelioma, a tumour derived from the peritoneal lining, that contains a homozygous point mutation within WT1. This mutation, within the putative transactivation domain, converts the protein from a transcriptional repressor of its target sequence to a transcriptional activator. The role of WT1 in normal development thus extends to diverse structures derived from embryonic mesoderm and disruption of WT1 function contributes to the onset of adult, as well as pediatric, tumours. PMID- 8401593 TI - A duplication in the L1CAM gene associated with X-linked hydrocephalus. AB - Recently, a mutation in the gene for the neural cell adhesion molecule L1CAM, located at chromosome Xq28, was found in a family with X-linked hydrocephalus (HSAS). However, as the L1CAM mutation could only be identified in one HSAS family, it remained unclear whether or not L1CAM was the gene responsible for HSAS. We have conducted a mutation analysis of L1CAM in 25 HSAS families. The mutation reported previously was not found in any of these families. In one family, however, a 1.3 kilobases (kb) genomic duplication was identified, cosegregating with HSAS and significantly changing the intracellular domain of the L1CAM protein. These results confirm that L1CAM is the HSAS gene. PMID- 8401594 TI - A trithorax-like gene is interrupted by chromosome 11q23 translocations in acute leukaemias. PMID- 8401595 TI - The neural basis of drug craving: an incentive-sensitization theory of addiction. AB - This paper presents a biopsychological theory of drug addiction, the 'Incentive Sensitization Theory'. The theory addresses three fundamental questions. The first is: why do addicts crave drugs? That is, what is the psychological and neurobiological basis of drug craving? The second is: why does drug craving persist even after long periods of abstinence? The third is whether 'wanting' drugs (drug craving) is attributable to 'liking' drugs (to the subjective pleasurable effects of drugs)? The theory posits the following. (1) Addictive drugs share the ability to enhance mesotelencephalic dopamine neurotransmission. (2) One psychological function of this neural system is to attribute 'incentive salience' to the perception and mental representation of events associated with activation of the system. Incentive salience is a psychological process that transforms the perception of stimuli, imbuing them with salience, making them attractive, 'wanted', incentive stimuli. (3) In some individuals the repeated use of addictive drugs produces incremental neuroadaptations in this neural system, rendering it increasingly and perhaps permanently, hypersensitive ('sensitized') to drugs and drug-associated stimuli. The sensitization of dopamine systems is gated by associative learning, which causes excessive incentive salience to be attributed to the act of drug taking and to stimuli associated with drug taking. It is specifically the sensitization of incentive salience, therefore, that transforms ordinary 'wanting' into excessive drug craving. (4) It is further proposed that sensitization of the neural systems responsible for incentive salience ('for wanting') can occur independently of changes in neural systems that mediate the subjective pleasurable effects of drugs (drug 'liking') and of neural systems that mediate withdrawal. Thus, sensitization of incentive salience can produce addictive behavior (compulsive drug seeking and drug taking) even if the expectation of drug pleasure or the aversive properties of withdrawal are diminished and even in the face of strong disincentives, including the loss of reputation, job, home and family. We review evidence for this view of addiction and discuss its implications for understanding the psychology and neurobiology of addiction. PMID- 8401596 TI - Amino acids as dietary excitotoxins: a contribution to understanding neurodegenerative disorders. AB - The possibility that some acidic amino acids occurring naturally or as additives in the diet can act as excitotoxins producing central nervous system pathology has been the subject of extensive debate in the last 20 years and is here reviewed. High doses of glutamate, aspartate or related excitatory amino acids given in isolation to neonatal rodents produce acute degeneration organs. Neuropathology resulting from consumption of glutamate or aspartate has not been described in man. Various unusual amino acids of plant origin can produce acute excitotoxic syndromes. In man domoate (consumed in mussels that have fed on (Nitschia pungens) can produce an acute syndrome associated with limbic system lesions and anterograde amnesia. Kainate and domoate produce similar syndromes in rodents; acromelate produces spinal pathology. The mechanisms and manifestations of chronic excitotoxicity are less clearly established. A combination of impaired energy metabolism or impaired buffering of calcium and free radicals and endogenous or exogenous excitotoxins may contribute to neuronal loss in human neurodegenerative disorders. PMID- 8401597 TI - Circadian rhythms. AB - Circadian rhythms are a ubiquitous adaptation of eukaryotic organisms to the most reliable and predictable of environmental changes, the daily cycles of light and temperature. Prominent daily rhythms in behavior, physiology, hormone levels and biochemistry (including gene expression) are not merely responses to these environmental cycles, however, but embody the organism's ability to keep and tell time. At the core of circadian systems is a mysterious mechanism, located in the brain (actually the suprachiasmatic nucleus of the hypothalamus) of mammals, but present even in unicellular organisms, that functions as a clock. This clock drives circadian rhythms. It is independent of, but remains responsive to, environmental cycles (especially light). The interest in temporal regulation--its organization, mechanism and consequences--unites investigators in diverse disciplines studying otherwise disparate systems. This diversity is reflected in the brief reviews that summarize the presentations at a meeting on circadian rhythms held in New York City on October 31, 1992. The meeting was sponsored by the Fondation pour l'Etude du Systeme Nerveux (FESN) and followed a larger meeting held 18 months earlier in Geneva, whose proceedings have been published (M. Zatz (Ed.), Report of the Ninth FESN Study Group on 'Circadian Rhythms', Discussions in Neuroscience, Vol. VIII, Nos. 2 + 3, Elsevier, Amsterdam, 1992). Some speakers described progress made in the interim, while others addressed aspects of the field not previously covered. PMID- 8401598 TI - Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth. AB - Advances in determination of polymer structure and in preservation of structure for electron microscopy provide the best view to date of how polysaccharides and structural proteins are organized into plant cell walls. The walls that form and partition dividing cells are modified chemically and structurally from the walls expanding to provide a cell with its functional form. In grasses, the chemical structure of the wall differs from that of all other flowering plant species that have been examined. Nevertheless, both types of wall must conform to the same physical laws. Cell expansion occurs via strictly regulated reorientation of each of the wall's components that first permits the wall to stretch in specific directions and then lock into final shape. This review integrates information on the chemical structure of individual polymers with data obtained from new techniques used to probe the arrangement of the polymers within the walls of individual cells. We provide structural models of two distinct types of walls in flowering plants consistent with the physical properties of the wall and its components. PMID- 8401599 TI - Gametophytic and sporophytic expression of an anther-specific Arabidopsis thaliana gene. AB - Genomic and cDNA clones of the anther-specific APG gene from Arabidopsis thaliana and Brassica napus, which encodes a novel proline-rich protein, were isolated and characterized. Southern blotting and Northern analysis of male fertile and cytoplasmic male sterile varieties of B. napus showed that the APG gene is present as a single copy in the Arabidopsis genome, and that the B. napus APG gene is a member of a small anther-specific gene family. Analysis of developmentally staged B. napus flower buds indicated that APG transcript is confined to the anther during the period of microspore development. Reporter gene fusions established that the APG promoter directs expression in a number of cell types in anthers of transformed plants. This expression is consistent with the temporal pattern of mRNA accumulation in B. napus buds and follows a complex developmental pattern. Most significantly, the promoter is active in both sporophytic and gametophytic cell types, with activity of the transgene in each cell type being delineated by various cytological markers. PMID- 8401600 TI - Molecular characterization of a 70 kDa heat-shock protein of bean mitochondria. AB - A bean cDNA clone that specifies a 70 kDa heat-shock protein (hsp70) has been isolated and sequenced. The nucleotide sequence analysis shows that the cDNA could encode a 72 kDa protein that is highly related to prokaryotic and mitochondrial members of the hsp70 family. The predicted protein was found to contain an amino-terminal extension typical of transit sequences. The in vitro transcription/translation product of the cDNA behaved as a 72 kDa polypeptide as predicted from the longest open reading frame. This polypeptide could be imported into isolated mitochondria and recovered as a 68 kDa product. The imported protein is identical in size to a mitochondrial protein that cross-reacts with hsp70-specific antibodies. The import data and Western blot analysis suggest that the cDNA clone encodes a mitochondrial member of the hsp70 family. Electrophoretic and immunoblot analysis reveal that the protein is loosely associated to the mitochondrial envelope and also exists as discrete soluble protein aggregates of about 270 and 420 kDa. Hsp70 of bean mitochondria can be in vitro phosphorylated on threonine residues in a calcium-dependent manner, and the modified protein was detected as an oligomer of about 160 kDa only. The data are discussed with respect to the chaperone function of hsp70 in mitochondria. PMID- 8401601 TI - Characterization of cDNAs for stylar transmitting tissue-specific proline-rich proteins in tobacco. AB - The pistil of flowers is a specialized organ which contains the female gametophytes and provides the structures necessary for pollination and fertilization. Pollen deposited on the stigmatic surface of a compatible plant germinates a pollen tube which penetrates the stigmatic papillae and grows intercellularly through the style towards the ovules in the ovary. Pollen tube growth is largely restricted to the transmitting tissue in the style. Therefore the stylar transmitting tissue is extremely important for the migration of the pollen cell towards the ovary. We have isolated two related cDNAs, transmitting tissue-specific (TTS)-1 and TTS-2, derived from two proline-rich protein (PRP) encoding mRNAs that accumulate specifically in the transmitting tissue of tobacco. The deduced PRP sequences share similarities with proline-rich cell wall glycoproteins found in a variety of plants. TTS-1 and TTS-2 mRNAs are induced in very young floral buds, accumulate most abundantly during the later stages of flower development when style elongation is the most rapid, and remain at relatively high levels at anthesis. These mRNAs become undetectable in maturing green fruits. In situ hybridization shows that TTS-1 and TTS-2 mRNA accumulation is restricted to the transmitting tissue of the style. The possible roles that these transmitting tissue-specific PRPs may play in maintaining the structural integrity of the style or in the function of this organ is discussed. PMID- 8401602 TI - Promoter and leader regions involved in the expression of the Arabidopsis ferredoxin A gene. AB - Ferredoxin is a nuclear-encoded protein that is involved in a variety of electron transfer reactions in both photosynthetic and non-photosynthetic plastids. We show here that the expression of the ferredoxin A gene (FedA) in Arabidopsis thaliana is light-regulated, with its mRNA level increased 4.5-fold by transfer of dark-grown seedlings to white light for 3 h. A portion of this light regulation is mediated by phytochrome through a very low fluence type of response. In addition, it is likely that another photoreceptor(s) is also involved. The FedA promoter confers a light- and tissue-regulated expression pattern when fused to the beta-glucuronidase (GUS) and luciferase reporter genes, indicating that the gene is transcriptionally regulated. No evidence of cis acting light-regulatory elements within the 5' untranslated leader region of the gene was detected. Nevertheless, elements within this leader are required for full activity since its deletion reduces expression both in the light and dark by 25-fold. This region includes a sequence, ACAAAA, which is also present in the 5' untranslated leader of the other three ferredoxin leaders that have been sequenced and in the leaders of 31 other plant genes or about 8% of all plant genes in the GenBank database. In addition to mediating light regulation, the FedA promoter also directs GUS expression in aerial tissues at 70-fold higher levels than in roots. GUS activity staining in aerial tissues is observed in both photosynthetic and non-photosynthetic cells. These data indicate that the FedA promoter also carries the information for expression of this gene in a highly tissue- and cell-specific manner. PMID- 8401603 TI - PCR-amplified microsatellites as markers in plant genetics. AB - In order to assess the feasibility of using microsatellites as markers in plant genetics, a survey of published DNA sequence data for presence, abundance and ubiquity in higher plants of all types of dinucleotide and trinucleotide repeats with a minimum number of 10 and 7 units, respectively, was conducted. This search revealed that such microsatellites are frequent and widely distributed; they were uncovered in 34 species, with a frequency of one every 50 kb. AT repeats were by far the most frequently observed class of dinucleotide microsatellites, whereas AC/TG repeats, which are common in animals, were observed only once. TAT repeats prevailed among trinucleotides. Polymerase chain reaction amplification of (AT)n and (TAT)n microsatellites in soybean (Glycine max (L.) Merr.) revealed that they are highly polymorphic, as a consequence of length variation, somatically stable and inherited in a co-dominant Mendelian manner. The abundance and amount of information derived from such markers, together with the ease by which they can be identified, make them ideal markers for plant genetic linkage and physical mapping, population studies and varietal identification. PMID- 8401604 TI - Detection of point mutations in the chloroplast genome by single-stranded conformation polymorphism analysis. AB - Single-stranded conformation polymorphism (SSCP) analysis was used to examine the mutations of the chloroplast 16S rRNA locus of streptomycin-resistant mutants in Nicotiana plumbaginifolia. DNA fragments of 121, 517, 968 and 1578 bp, each possessing a known point mutation, were generated by polymerase chain reaction (PCR). The resulting fragments were denatured and separated by nondenaturing polyacrylamide gel electrophoresis. Compared to the patterns of the wild-type DNA fragments, the bands of the single-stranded DNA fragments of 121 and 517 bp with base changes were shifted. However, no pattern variations were detected from the DNA fragments of 968 and 1578 bp generated from both wild-type and mutants. PMID- 8401605 TI - Plant chitinases. PMID- 8401606 TI - Isolation and characterization of male flower cDNAs from maize. AB - Differential screening of two libraries made from whole, immature maize tassels was used to isolate six cDNAs which show enhanced levels of expression in male flowers. MFS1, MFS2, MFS4, MFS10 and MFS18, which were isolated from a 5 cm tassel library, are expressed throughout tassel growth up until mature pollen is produced in the anthers. MFS14, which was isolated from a 10-12 cm tassel library, has a narrower window of expression associated with microsporogenesis and declines as mature pollen is produced. MFS18 mRNA accumulates in the glumes and in anther walls, paleas and lemmas of mature florets. MFS18 mRNA is particularly associated with the vascular bundle in the glumes and encodes a polypeptide of 12 kDa, rich in glycine, proline and serine that has similarities with other plant structural proteins. In contrast, MFS14 mRNA accumulates in the tapetum and encodes a polypeptide of 13 kDa that is rich in alanine. The MFS14 and MFS18 proteins are basic (isoelectric points of 11.56 and 9.54, respectively) and both have hydrophobic N-termini which display all the characteristics of signal peptides, indicating that these proteins may be secreted. PMID- 8401607 TI - Expression of myb-related genes in the moss, Physcomitrella patens. AB - Three cDNA clones encoding proteins containing a myb-related DNA binding domain have been isolated from a cDNA library prepared from protonemal tissue of the moss, Physcomitrella patens. The three cDNA clones between them encode two different classes of myb-like proteins, termed Pp1 and Pp2, that, outside of the myb domain, show no regions of significant homology. Acidic domains, capable of forming alpha-helical structures, are present in the carboxy-termini of the derived amino acid sequences from Pp1 and Pp2cDNAs suggesting that, like other myb genes, these proteins probably function as transcriptional activators. In contrast to other plants, where extensive myb-related gene families are present in the genome, a relatively small family is present in P. patens. Analyses of transcript levels during development of P. patens showed that maximum levels of transcription of the two genes occurred in young wild-type protonemal tissue that correlated with the time of maximum mitotic index. A decline in the expression of both genes occurs with increasing age of the wild-type tissue. Aberrant levels of expression of the two genes were observed in developmental mutants of P. patens which, as well as carrying specific morphological mutations, have greatly retarded protonemal growth rates. Transformation of wild-type P. patens with antisense constructs derived from Pp1 and Pp2 cDNA clones led to a dramatically reduced frequency of transformants when the expression of the reporter gene within the constructs was selected. Taken together, the data strongly suggest that expression of Pp1 and Pp2 is essential for cell growth during normal gametophytic development of P. patens. PMID- 8401608 TI - Mechanosensory calcium-selective cation channels in epidermal cells. AB - This paper explores the properties and likely functions of an epidermal Ca(2+) selective cation channel complex activated by tension. As many as eight or nine linked or linkable equivalent conductance units or co-channels can open together. Open time for co-channel quadruplets and quintuplets tends to be relatively long with millimolar Mg2+ (but not millimolar Ca2+) at the cytosolic face of excised plasma membrane. Sensitivity to tension is regulated by transmembrane voltage and temperature. Under some circumstances channel activity is synchronized in rhythmic pulses. Certain lanthanides and a cytoskeleton-disturbing herbicide that inhibit gravitropic reception act on the channel system at low concentrations. Specifically, ethyl-N-phenylcarbamate promotes tension-dependent activity at micromolar levels. With moderate suction, Gd3+ provided at about 0.5 microM at the extracellular face of the membrane promotes for several seconds but may then become inhibitory. Provision at 1-2 microM promotes and subsequently inhibits more vigorously (often abruptly and totally), and at high levels inhibits immediately. La3+, a poor gravitropic inhibitor, acts similarly but much more gradually and only at much higher concentrations. These properties, particularly these susceptibilities to modulation, indicate that in vivo the mechanosensitive channel must be mechanosensory and mechanoregulatory. It could serve to transduce the shear forces generated in the integrated wall-membrane-cytoskeleton system during turgor changes and cell expansion as well as transducing the stresses induced by gravity, touch and flexure. In so far as such transduction is modulated by voltage and temperature, the channels would also be sensors for these modalities as long as the wall-membrane-cytoskeleton system experiences mechanical stress. PMID- 8401609 TI - Cotton fiber annexins: a potential role in the regulation of callose synthase. AB - Cotton fibers contain a characteristic set of proteins which interact with plasma membranes in a Ca(2+)-dependent manner. The association of these proteins with the membrane is correlated with a reduced level of UDP-glucose: (1-->3)-beta glucan (callose) synthase activity. Analysis of the proteins released from membranes by EDTA treatment shows that the most abundant proteins comprise a family of at least three polypeptides (p34) which resemble annexins. This resemblance includes similarity in size (about 34 kDa), sequence homology, Ca(2+) dependent precipitation or interaction with the plasma membrane, and ability to serve as a substrate for phosphorylation by endogenous protein kinase(s) which also bind to the membranes in a Ca(2+)-dependent manner. A purified fraction of these annexins binds to, and inhibits, the activity of a partially purified cotton fiber callose synthase. These findings suggest that one possible function of annexin(s) in plants is to modulate the activity and/or localization of callose synthase. PMID- 8401610 TI - Mobility of the maize transposable element En/Spm in Arabidopsis thaliana. AB - The autonomous element En-1 of the maize En/Spm transposable element system is capable of frequent somatic and germinal excision in the heterologous host Arabidopsis thaliana. The pattern of En-homologous transcripts generated in transgenic Arabidopsis resembles En transcription in maize. An excision reporter construct based on NPT-II gene (pKEn2) can be used reliably for the isolation of En-1 germinal revertants by seed germination on kanamycin-containing medium. Re insertion after germinal excision is apparently frequent. A dSpm receptor element can be efficiently trans-activated in Arabidopsis either by En-1 or by expressing cDNAs of tnpA and tnpD. Excision and re-insertion of En/Spm take place with similar characteristics as in maize. This is the first description of En/Spm transposition in Arabidopsis and the parameters analysed here suggest that transposon tagging with En should be feasible in this species. PMID- 8401611 TI - Differential expression of the rolA plant oncogene and its effect on tobacco development. AB - The rolA gene not only provokes the abnormal traits of wrinkled leaves, shortened internodes, and deficient root growth in tobacco, but also induction of roots in leaf disks in hormone-free medium. The rolA mRNA is most abundant in the stem of transgenic tobacco, accumulating at fivefold and 50-fold lower levels in the leaf and the root, respectively. Promoter deletion mutants were constructed and the rolA-induced abnormal traits were exploited to identify promoter regulatory regions. The removal of specific promoter domains provokes the appearance of new patterns of rolA transcripts in leaves, stems and roots. These new patterns are correlated with the disappearance of specific abnormal trait, thereby identifying three functional domains. Domain A includes a mosaic motif composed of a putative GT-2 binding site and DUE-NDE elements. This domain positively regulates rolA gene expression in the leaf and negatively regulates it in the stem and root. Domains B and C, which contain sequences homologous to the OCS enhancer and to ASF-1 binding sites, respectively, generally activate the rolA gene expression in all tissues. The possible function of these elements on the regulation of the rolA gene is discussed. PMID- 8401612 TI - Light-regulated expression of the Arabidopsis thaliana ferredoxin A gene involves both transcriptional and post-transcriptional processes. AB - Ferredoxin is part of the photosynthetic apparatus of the chloroplast and is encoded in the nucleus. In Arabidopsis thaliana expression of the ferredoxin A gene is influenced by both the presence of chloroplasts and light. Tobacco plants transformed with a ferredoxin promoter-GUS fusion gene showed a tissue-specific and light-dependent expression pattern. The effect of light on the expression of the fusion gene in transgenic seedlings was only two- to fourfold, which is less pronounced than the 20-fold effect in Arabidopsis itself. Run-on transcription assays with nuclei isolated from Arabidopsis revealed a twofold modulation of transcriptional activity of the ferredoxin A gene under the influence of light. These results suggest the involvement of post-transcriptional processes in light regulated gene expression. A ferredoxin promoter deletion series ranging from 1205 to -143 was studied. All but the smallest deletion construct (at position 143 relative to the translation start site) showed comparable expression levels in mature leaves, suggesting the presence of a positive regulating element between -269 and -143. The same pattern of tissue specificity was found in all promoter deletions studied. Expression of the fusion genes is high in all chloroplast-containing cells: mesophyll, chlorenchyma, paravascular tissue, epidermal and stomatal guard cells and trichomes. Transgenic seedlings treated with norflurazon, which blocks the development of green chloroplasts, showed a two- to fourfold reduction in GUS expression for all constructs. In Arabidopsis seedlings the effect of norflurazon on the expression of the ferredoxin A was eightfold. This again can be explained by the need for post-transcriptional processes of the regulated gene expression of Arabidopsis ferredoxin A. PMID- 8401613 TI - Purification and biochemical characterization of proteins which bind to the H-box cis-element implicated in transcriptional activation of plant defense genes. AB - The H-box (CCTACC(N)7CT(N)4A), which occurs three times within the -154 to -42 region of the bean chalcone synthase chs15 promoter, is important for developmental regulation of chs15, and induction of chs15 and coordinately regulated defense genes by elicitors and other stress stimuli. Two protein factors, KAP-1 and KAP-2, which recognize conserved features in the H-box motif, were purified from bean cell suspension cultures by a combination of ion exchange chromatography and DNA affinity chromatography. KAP-1 is a 97 kDa polypeptide, whereas KAP-2 comprises two polypeptides of 76 and 56 kDa. KAP-1 and KAP-2 also differ in the sensitivity of their DNA-bound forms to trypsin. Dephosphorylation of KAP-1 or KAP-2 affects the mobility of the protein/H-box binding complex in gel shift assays but does not inhibit DNA binding. Elicitation of bean cell suspensions with glutathione does not affect the total cellular activities of KAP 1 or KAP-2, but causes a rapid increase in the specific activities of both factors in the nuclear fraction, consistent with a role for these factors in the signal pathway for elicitor induction of chs15 and related defense genes. PMID- 8401614 TI - Transcription of two members of a gene family encoding phenylalanine ammonia lyase leads to remarkably different cell specificities and induction patterns. AB - Phenylalanine ammonia-lyase (PAL) catalyses the first committed step in the biosynthesis of phenylpropanoids, which perform a variety of functions in plant development and in their interactions with the environment. French bean contains a small family of genes encoding PAL and two of these genes, PAL2 and PAL3, have been shown to be differentially expressed at the mRNA level in bean tissues. The transcriptional activities of the PAL2 and PAL3 genes have been investigated by fusing their promoters to the reporter gene beta-glucuronidase (GUS) and transforming these constructs into Arabidopsis, potato and tobacco. The PAL2- and PAL3-GUS constructs exhibited different spatial and temporal patterns of expression during development and in response to environmental stimuli. The consistency of these data with previous mRNA analysis in bean suggests that the differential expression of these two PAL genes is, at least in part, a function of their promoter activities. New patterns of PAL2 and PAL3 promoter activities were also characterized. Some species-specific differences in GUS expression were observed and these may reflect differences in phenylpropanoid metabolism or the signals that modulate PAL gene transcription. PMID- 8401615 TI - The MsK family of alfalfa protein kinase genes encodes homologues of shaggy/glycogen synthase kinase-3 and shows differential expression patterns in plant organs and development. AB - This paper reports on the isolation of a novel class of plant serine/threonine protein kinase genes, MsK-1, MsK-2 and MsK-3. They belong to the superfamily of cdc2-like genes, but show highest identity to the Drosophila shaggy and rat GSK-3 proteins (65-70%). All of these kinases share a highly conserved catalytic protein kinase domain. Different amino-terminal extensions distinguish the different proteins. The different plant kinases do not originate from differential processing of the same gene as is found for shaggy, but are encoded by different members of a gene family. Similarly to the shaggy kinases, the plant kinases show different organ-specific and stage-specific developmental expression patterns. Since the shaggy kinases play an important role in intercellular communication in Drosophila development, the MsK kinases are expected to perform a similar function in plants. PMID- 8401616 TI - Localization of vicilin genes via polymerase chain reaction on microisolated field bean chromosomes. AB - A new technique is reported for the physical mapping of low copy DNA sequences on plant chromosomes. Individual chromosomes were microisolated and their DNA used as the target for the polymerase chain reaction in order to identify the chromosome carrying a specific gene sequence. The use of defined translocation chromosomes further refined the resolution of the method to a subchromosomal level. To demonstrate the applicability of the procedure genes have been localized coding for vicilin seed storage proteins on the field bean Vicia faba L. in a region which includes the centromere and the proximal parts of the short and the long arms of chromosome II. PMID- 8401617 TI - [Oral ofloxacin versus intramuscular ceftriaxone in antibiotic prophylaxis in transurethral prostate resection]. AB - This randomized prospective study evaluated the safety and efficacy of a single oral dose of OFL compared to a single parenteral dose of CTRX prior to TURP. 191 patients (mean age: 68.7 +/- 6.2 years) with bacterial free urine before surgery were enrolled and received either OFL: 400 mg per os (n = 95) or CTRX: 1 g intrasmuscularly (n = 96) at the pre-anaesthetic medication time (two hours prior TURP). Two urine cultures were obtained: on the day of the patient's discharge and within one month after surgery. Blood cultures were performed in case of temperature > 38 degrees 5C. Treatment failure was defined as bacteriuria > 10(5) CFU/ml and/or in case of positive blood culture after surgery. 182 patients were evaluable for efficacy. They were similar with respect to age, prostatic resection, histology, duration of post operative catheterization in the two treatment groups. On discharge from the department, 93.2% of the patients in the OFL group had sterile urines versus 94.6% in the CTRX group. In failure, causative pathogens were in the OFL group: 3 enterococcus, 1 acinetobacter, 1 staphylococcus, 1 citrobacter; in the CTRX group: 1 acinetobacter, 2 citrobacters, 1 enterococcus, 1 streptococcus. No bacteremia occurred. After one month of follow-up, 171 patients were evaluable (success on discharge from the department): the rates of success were: 93.5% in the OFL group and 90.3% in the CTRX group. Tolerance was good in both groups. These two drugs are as effective and as well tolerated for prevention of post operative UTI in TURP. But Ofloxacin is cost effective and simplest for use. PMID- 8401618 TI - [The value of antibiotic prophylaxis in transurethral resection of bladder tumors. Apropos of 61 cases]. AB - In order to define the value of antibiotic prophylaxis, we conducted a prospective, randomised, double-blind, placebo-controlled study in 61 patients undergoing transurethral resection (TUR) of a bladder lesion suspected to be neoplastic. 32 patients received a single dose of 800 mg of pefloxacin at the time of anaesthetic induction and 29 patients received placebo. All patients had sterile preoperative urine and none had received any antibiotics during the fortnight preceding the operation. 3 patients in the pefloxacin group (9.4%) developed postoperative bacteriuria versus 7 in the placebo group (24.1%) (no statistically significant difference). No patient developed symptomatic urinary tract infection. We conclude that antibiotic prophylaxis is not indicated during TUR for bladder tumours. PMID- 8401619 TI - [Survey of instillation methods of BCG in the preventive treatment of superficial tumors of the bladder. Subcommittee of Superficial Tumors of the Bladder, the Committee of Cancerology of the French Association of Urology]. AB - In an attempt to appreciate urological practices concerning BCG instillations a study was performed with members of university urological centers. Questions concerned instillation modalities, persons performing instillations and criteria for survey. Main differences concerned doses (low doses versus high doses) and duration of treatment. These results suggest the need of development of new parameters for the rationalisation of this treatment. PMID- 8401620 TI - [The prevention of the recurrence of stage pTa bladder tumors using intravesical instillation of BCG]. AB - 45 patients were included in a retrospective study designed to assess the long term efficacy and tolerance of immunotherapy by intravesical BCG instillation in the treatment of stage pTa bladder tumours (mean follow-up 52.7 months, range: 12 87 months). 41 patients (91% responded to immunotherapy: 24 (53%) did not develop anu recurrences with a mean follow-up of 49.4 months (range: 19-87 months) and 17 (37.7%) were improved by treatment as the interval between recurrences was significantly increased. 4 patients (8.8%) failed to respond to BCG therapy with progression to stage pTa G3 vesical papillomatosis in one patient. The local tolerance was poor for twelve patients (26.6%), requiring discontinuation or spacing of the instillations. Other complications such as prostatitis and intense fever were observed. Three patients had persistent disturbances of micturition following treatment in the form of urinary frequency due to the decreased bladder volume. Neither the grade, nor the frequency of recurrences prior to treatment, nor the solitary or multifocal nature, nor the site of the initial tumour appeared to influence the response to BCG therapy and did not constitute independent individual prognostic indicators. PMID- 8401621 TI - [Treatment of stage Ta,T1 and Tis bladder tumors using Calmette-Guerin bacillus vaccine]. AB - The authors report a series of 71 patients (sex ratio: 1F/4M, mean age: 68 years) with stage Ta (n = 20), T1a (n = 32), T1b (n = 14) and Tis (n = 5) bladder tumours treated by endoscopic resection followed by a course of intravesical BCG instillation (120 mg/week for 6 weeks). The mean follow-up was 15 months (3-36 months). The overall recurrence rate was 42%. A recurrence occurred in 50% of Ta (median time to recurrence: 10.1 months), 32% of T1a (median: 5.8 months), 65% of T1b (median: 7.3 months) and 20% of isolated Tis (median: 7 months). Disease progression was observed in 9% of stage T1 tumours. The following risk factors for recurrence were identified: stage T1b (p = 0.05), multifocal tumours (p = 0.05), resistance to previous chemotherapy (mitomycin C) (p = 0.001) and association with Tis for stages T1 (p < 0.02). The following risk factors for disease progression were identified: stage T1b (p < 0.001), grade III for stage T1 (p = 0.05) and association with Tis (p < 0.05). Ten patients (14%) developed transient BCGitis. BCG was found to be effective in the prophylaxis of recurrence of stage Ta, T1 and Tis bladder tumours. This treatment is proposed for recurrent stage Ta grade II and III tumours and stage T1 tumours in the presence of recognised risk factors. The high risk of progression for stage T1b grade III tumours associated with Tis demands rigorous surveillance. PMID- 8401622 TI - [Urothelial tumors of the bladder in patients less than 25 years of age]. AB - During the period of October 1981 to October 1990, we diagnosed and treated 7 patients (5 males and 2 females) under 25 years old (mean age 21.2) with transitional cell carcinoma of the bladder. The diagnosis and the treatment were the same as in the older age group. The tumors were of low grade and low stage and presented a recurrence rate of 28.5%. The tumors were of favourable prognosis but the follow-up care of these patients should be vigilant and persistent. PMID- 8401623 TI - [Repeated renal transplantation]. AB - From October 1987 to June 1992, 359 renal transplantations were performed, while, over the same period, 25 patients with a follow-up of more than six months underwent repeat renal transplantation: 23 for a second transplant and 2 for a third transplant. The initial disease was glomerular for 17 patients and interstitial for 6 patients. The mean age of the patients at the time of the repeat transplantation was 36.9 years (range: 20 to 53 years). The mean survival after the first transplantation was 3 years and 10 months (range: 1 week to 10 years). The reasons for loss of the first transplant can be classified as follows: acute rejection (n = 5), chronic rejection (n = 14), surgical failure (n = 5) or sepsis (n = 1). 9 patients received conventional immunosuppressant therapy, while 16 patients (64%) received four-drug therapy including cyclosporin. The actuarial one-year survival of the patients and the transplants was 100% and 92%, respectively. The mean serum creatinine was 136.4 +/- 65 mumol/l (range: 59 to 298 mumol/l). Ten patients developed rejection after a mean of 18.6 days (range: 6 to 30 days) and 2 patients suffered from 2 episodes of rejection within 4 months. These results illustrate the low postoperative surgical and immunological complication rate in this group of patients whose long term results are at least comparable to those of first transplantations in our group. PMID- 8401624 TI - [Renal transplantectomy: surgical technics and results. Apropos of 60 patients]. AB - Nephrectomy of a non-functioning renal transplant after renal transplantation is a delicate operation which the authors evaluate on the basis of a retrospective analysis of 62 patients. A review of the literature helps to more clearly define the indications, optimal time of the operation and the operative technique. A subcapsular approach, by simplifying the operation, constitutes an important factor in reducing the operative morbidity in the late forms. "Early" transplantectomies are often more serious because of the recently operated patient's fragile status and immunosuppression, responsible for frequent infectious complications. Transplantectomy should therefore not be delayed once the permanent loss of transplant function has been confirmed. PMID- 8401625 TI - [The 1st 100 kidney transplants performed in Tunisia and their surgical complications]. AB - The authors report their experience of renal transplantation based on the first 100 transplantations performed in Tunisia. The absence of legislation concerning organ harvesting from brain dead subjects has favoured renal transplantation from living donors. 20 surgical complications are reported and analysed. PMID- 8401626 TI - [Instantaneous pressure-flow relations in a continuous flow in a collapsible tube: impact on the evaluation of urethral resistance]. AB - The urethra is a collapsed tube in the absence of micturition. Consequently, the evaluation of instantaneous urethral resistance by the pressure/flow relation (R = P/Q2) derived from Bernoulli's equation has been criticised for not taking into account the opening pressure or at least the fact that flow must maintain patency of the urethra during voiding. The objective of this study was to determine whether such a criticism is justified in terms of fluid mechanics. A study of the pressure/flow relation was performed in a soft tube, collapsed by various peritubular pressures, which subsequently opened in response to flow. Experiments demonstrated that the pressure/flow relation, the instantaneous resistance to flow and the pressure drop were identical when the tube reached the same calibre, despite different peritubular pressures. The same pressure therefore propels the flow and maintains the patency of the tube. Consequently, it would be unreasonable to expect to improve the evaluation of instantaneous urethral resistance by subtracting an opening pressure from the maximal bladder pressure. The formula R = P/Q2 is valid, experimentally, and complies with the principles of fluid mechanics. In theory and in practice, it constitutes a good method of evaluation of instantaneous resistance. PMID- 8401627 TI - [Percutaneous surgery for lithiasis: results and perspectives. Apropos of 390 operations]. AB - Percutaneous nephrolithotomy (PCNL) has radically changed the treatment of renal stones. The indications for this technique have been modified by the development and refinement of extracorporeal lithotripsy (ECL). The authors present their experience of 390 PCNL performed between 1984 and 1991, for solitary stones in 75% of cases. There were only 11 cases of failed puncture (3.8%). The overall morbidity was 18% with only 4.4% of major complications, i.e. life threatening or requiring reoperation. There were two deaths (0.5%). Stones were completely eliminated in 80.25% of patients, after a second operation (PCNL or other technique) in 32 cases (11%). 45 staghorn calculi were treated with 11% of complications and a 51% complete cure rate. The mean hospital stay was 6 days (2 to 30). PCNL is a safe and reliable technique. Its indications have decreased (6.5% of our patients treated for renal stones), but it still retains a place in the therapeutic approach to patients with renal stones. PMID- 8401628 TI - [Urologic manifestation of Waldenstrom's disease. Apropos of a case with pyelo ureteral location]. AB - The authors report a case of Waldenstrom's disease in which the initial staging assessment reveals an isolated tumour of the left upper urinary tract. Combination chemotherapy (6 courses of protocol M2) induced a marked reduction in the monoclonal IgM peak and in the tumour mass. In the light of this atypical case, which emphasises the diversity of the potential sites of Waldenstrom's macroglobulinaemia, the authors review its potential for progression and the various therapeutic modalities available. PMID- 8401629 TI - [Drainage of a lymphocele after kidney transplantation, using laparoscopy]. AB - The authors report a case of transperitoneal laparoscopic drainage of a recurrent, compressive lymphocele occurring 3 months after cadaver renal transplantation. The technique consists of creating a transperitoneal breach and resecting the walls of the lymphocele to ensure internal drainage followed by suture of a slip of greater omentum over the opened lymphatic cavity. Preoperative aspiration and computed tomography precisely defined the site of the lymphocele in relation to urinary and vascular structures, confirmed the lymphatic nature of the collection and excluded the presence of infection. This technique has the same indications as classical surgical internal drainage and can be used to easily perform the same procedures. Because of its simplicity and low morbidity, laparoscopic drainage can be proposed as first-line treatment for large, symptomatic and recurrent posttransplantation lymphoceles, in the absence of infection and provided the lymphocele is in an accessible site and the operator has a sufficient experience of laparoscopic techniques. PMID- 8401630 TI - [Primary bladder signet ring cell adenocarcinoma. Apropos of 1 case. Review of the literature]. AB - The authors report a new case of signet ring cell carcinoma of bladder whose characteristic findings are the parietal infiltration and the rapidly fatal course. Based on a review of the literature, they define the histological features of this very rare tumour (less than 20 documented cases have been reported). The pluripotent behaviour of urothelial cells appears to be the essential element in the histogenesis of this tumour type, which is very similar to gastric linitis plastica. PMID- 8401631 TI - Superficial bladder carcinoma. Factors affecting the natural history. AB - The natural history of superficial bladder carcinoma has an important bearing on the outcome of response to intravesical chemotherapy or immunotherapy. It may vary in different parts of the world, but it is essential that urologists know within their own practice how tumours respond to the initial transurethral resection. Many factors influence the natural history of superficial bladder carcinoma, and some, such as poor-grade, large tumour at presentation, smoking history, poor response to initial resection, and dysplasia or carcinoma in situ, should alert the urologist to introduce intravesical therapy at an early stage. PMID- 8401632 TI - Prognostic factors in superficial bladder cancer. AB - A large body of evidence suggests that intravesical prophylaxis using bacillus Calmette-Guerin can favorably alter the natural history of superficial bladder cancer. The search for prognostic factors impacting the clinical alternative between conservative and radical treatment has been the subject of numerous efforts and remains a major objective in improving the carcinological results and the quality of life patients with superficial bladder cancer. In this regard, information is available on three levels: clinicopathological parameters, quantitative analysis, and biological markers. Recent advances in as well as the limitations and pitfalls of the use of three kinds of information in prognostication are reviewed. The proper use of prognostic factors will modify our attitude toward superficial bladder cancer in the near future. PMID- 8401633 TI - The evaluation and follow-up of patients with Ta, Tcis, and T1 bladder cancer. AB - Endoscopic evaluation and transurethral resection are the most important steps in the management of patients with bladder cancer. While multifocality is primarily a prognostic factor with respect to tumor recurrence, tumor grade and depth of invasion are prognostic regarding progression in stage. Transurethral resection biopsies of the prostate should always been obtained in patients with high grade tumors. Follow-up of patients should be individualized: patients with risk for progression, but not necessarily for recurrence, should be followed more closely than patients with low-grade tumors and low risk for progression. In this setting cytology is especially helpful because most high-grade tumors will be detected by cytology. Intravesical therapy is indicated in patients with high-grade tumors, carcinoma in-situ and multiple low grade tumors. While there is a growing body of evidence that BCG is superior to intravesical chemotherapy, one also has to take into account the more frequent and more serious side effects of BCG when compared with mitomycin C or thiotepa. PMID- 8401634 TI - Flow cytometry versus urinary cytology in the diagnosis and follow-up of bladder tumors: critical review of a 5-year experience. AB - The authors report the results of a 5-year experience with flow cytometry (FCM) in the diagnosis and evaluation of bladder tumors. FCM was applied to 400 patients (225 without a urinary tumor and 175 with a past or recent bladder tumor). For the patients without a tumor, bladder-irrigation-fluid FCM was positive in 22% of samples vs 1% of those tested with conventional cytology. The high rate of false-positive results may have been due to an imperfect quality of the samples, to the staining procedure and to the high amount of squamous cells. For 72 selected tumors in which the bladder washing was performed under specific requirements, the detection rate of FCM was lower than that of conventional cytology, including G1-grade tumors. A comparison between bladder-irrigation fluid FCM and disaggregated-biopsy FCM revealed a 22% rate of discordant results. From these data it does not seem desirable tu use bladder-irrigation FCM instead of conventional cytology in routine urologic examinations. The use of this technique must be more selective in bladder tumor evaluation. PMID- 8401635 TI - Lamina propria microinvasion of bladder tumors, incidence on stage allocation (pTa vs pT1): recommended approach. Pathologists of the French Association of Urology Cancer Committee. AB - A total of 100 bladder cancers posing a diagnostic problem as to invasion of the lamina propria were reevaluated by several pathologists. Agreement was reached in 90 cases after 4 assessments, but no agreement was obtained in the remaining 10 cases. In general, errors were due to overevaluation of microinvasion; only half of the "pT 1 tumors" remained classified as microinvasive pT 1 tumors. We propose that in the assessment presented to the clinician, there should be a clear distinction between pTa tumors with a doubt as to microinvasion, microinvasive pT 1 tumors, and highly invasive pT 1 tumors. PMID- 8401636 TI - Mechanisms of action of bacillus Calmette-Guerin in the treatment of superficial bladder cancer. AB - Local immunotherapy with bacillus Calmette-Guerin (BCG) is an effective treatment to prevent recurrence and progression of superficial bladder cancer, but the antitumor mechanism of action of BCG remains unclear. There are some experimental and clinical data suggesting that BCG antigens are processed not only by immunocompetent cells but also by urothelial cells and tumor cells. The foreign antigen may be presented at the cell surface by major histocompatibility complex (MHC) class II molecules and recognized by CD4 cells. The cytotoxic effect could result from the direct activity of CD4 cells or from the cytotoxic effect of released cytokines and the activation of other cytotoxic cells [cytolytic T lymphocytes CTLs; CD8 cells), macrophages, natural killer (NK) or lymphokine activated killer (LAK) cells]. These mechanisms are also involved in tumor rejection, and the identification of some specific tumor rejection antigens presented to specific CTLs could provide new therapeutic approaches. PMID- 8401637 TI - Adjuvant chemotherapy following radical cystectomy. AB - The optimal management of invasive bladder cancer remains controversial. Combination chemotherapy regimens have been increasingly advocated in association with definitive local therapy in an attempt at eradicating micrometastases and reducing the risk of recurrence. Chemotherapy used after radical cystectomy is referred to as "adjuvant" therapy. The term adjuvant is occasionally used for chemotherapy following an aggressive transurethral resection (TUR). Decisions concerning individual patients must be made after careful examination of the histologic specimen and cognizance of the known relapse rates per pathologic stage. No randomized trial has reported results on patients with pT 2 and pT 3a tumors. Studies have not clearly proven any advantage for adjuvant therapy based on muscle infiltration alone (pT 2, pT 3a). For patients with minimal extravesical extension (pT 3b), additional therapy may be useful. For patients with nodal metastases (pN+) and direct extension into the adjacent viscera (pT 4), the data suggest a trend toward improved survival. These patients may benefit from adjuvant chemotherapy. The difficulties of clinical trials following cystectomy, conducted in an elderly patient population, done over a long period, with changing standards of effective chemotherapeutic regimens are described. Adjuvant therapy following cystectomy or an aggressive TUR remains investigational and its benefit has not yet been proven. Patients should be entered into well-designed and carefully conducted clinical trials to evaluate the role of chemotherapy in invasive bladder cancer. PMID- 8401638 TI - Retroperitoneal laparoscopic ureterolithotomy. AB - Retroperitoneal laparoscopic ureterolithotomy was successfully performed in five patients with calculi impacted in the upper and middle ureter, using the recently described method of retroperitoneoscopy. All of the patients were discharged the following day but could have been treated as day cases. PMID- 8401639 TI - Clinical experience with multiagent intravesical therapy in interstitial cystitis patients unresponsive to single-agent therapy. AB - A total of 25 patients with the diagnosis of interstitial cystitis (IC) were involved in this study. All patients had been previously diagnosed with interstitial cystitis and had received treatment with single intravesical agents. Patients were divided into two groups according to their bladder capacity. The bladder capacity was > 350 ml in group I patients and < 350 ml in group II patients. For our study, dimethylsulfoxide (DMSO), methylprednisolone, and heparin sulfate were given every week for a total of 6 weeks. When symptoms recurred, supportive oral pharmacotherapy consisting of anticholinergics and/or tricyclic antidepressants was given. Under anesthesia, patients in group I showed a 99% increase in their bladder capacity; whereas those in group II showed an increase of only 19%. Cystoscopically, Hunner's ulcers were present in 60% of the group II patients but were seen in only 5% of the group I patients. Histopathological examination showed that the inflammatory changes were more frequent and severe in group II than in group I. Mast-cell hyperplasia was present in 100% of the patients in group II, versus only 61% of those in group I. A total of 23 patients (92%) achieved an initial remission averaging 8.1 months. In all, 9 patients (35%) had 1 or more relapses, and all achieved a subsequent remission averaging 8 months. By this combined multiagent approach, the majority of patients with IC obtained relief from their incapacitating symptoms and were socially rehabilitated. PMID- 8401640 TI - The electrified catheter. Role in sterilizing urine and decreasing bacteriuria. AB - A catheter was designed to decrease the incidence of catheter-induced urinary infections. A randomized, controlled study was performed to assess its safety and efficacy. The study comprised 24 patients subjected to hemorrhoidectomy. The criteria for entry were a preoperative urinary pathogen count below 10(5) colony forming units/ml urine as well as postoperative urinary retention that did not respond to conservative measures. The 24 patients were randomly assigned to treatment using a Nelaton catheter in 12 patients and an electrified catheter in the remaining 12. The catheters were left in place for 3 days, during which cephalosporin was given parenterally. The electrified catheter (EC) carried two silver-silver chloride electrodes. During the 3 days of catheterization and for 4 days thereafter, daily urine samples were cultured. The criterion for catheter induced urinary infection was a pathogen count above 10(5) colony-forming units/ml urine. No complication was encountered from catheterization. Electrodes were not broken. A bacterial count above 10(5) colony-forming units was found in 7 of 12 patients in the Nelaton catheter group, and a count below 10(5) units was determined in the 12 patients in the EC group. The study demonstrated that the EC can decrease the incidence of catheter-induced urinary infections. The EC is safe, cost-effective, and without complications. PMID- 8401641 TI - Early identification of hearing impairment in infants and young children. AB - The National Institutes of Health Consensus Development Conference on Early Identification of Hearing Impairment was convened to address (1) the advantages of early identification of hearing impairment and the consequences of late identification of hearing impairment; (2) the issue of which children should be screened for hearing impairment and when; (3) the advantages and disadvantages of current screening methods; (4) the question of which model for hearing screening and followup is preferred; and (5) future directions for research in diagnosis and management of hearing impairment in infants and young children. Following 2 days of presentations by experts and discussion by the audience, a consensus panel weighed the evidence and prepared their consensus statement. Among their findings, the panel concluded that (1) all infants admitted to the neonatal intensive care unit be screened for hearing loss prior to discharge; (2) universal screening be implemented for all infants within the first 3 months of life; (3) the preferred model for screening should begin with an evoked otoacoustic emissions test and should be followed by an auditory brainstem response test for all infants who fail the evoked otoacoustic emissions test; (4) comprehensive intervention and management programs must be an integral part of a universal screening program; (5) universal neonatal screening should not be a replacement for ongoing surveillance throughout infancy and early childhood; and (6) education of primary caregivers and primary health care providers on the early signs of hearing impairment is essential. PMID- 8401642 TI - [The role of peptidases in protein assimilation (a review of the current data)]. PMID- 8401643 TI - [The role of induced autolysis during digestion in fishes]. AB - A considerable activity of digestive hydrolases (carbohydrases in particular) was found in fish during feeding. The enzyme activity increases 2-6-fold at low values of the pH due, probably, to autolysis. This mechanism is important for the digestion in fish and other wild animals. PMID- 8401644 TI - [The enzyme topography of the small intestine in Arctic foxes and mink]. AB - High level of activity of some digestive enzymes in the distal part of the intestine is characteristic for predatory animals. The role of spatial redistribution of the enzyme activity during the process of dietary adaptation, is discussed. PMID- 8401645 TI - [Disordered permeability of the gastrointestinal tract barrier for macromolecules and the possibilities for its experimental dietetic correction]. AB - Macromolecular absorption of food and microbial antigens being enhanced in the intestine under pathological conditions may well be the cause of such diseases as food allergy, coeliac disease, Crohn's disease, atopic eczema, etc. The polyethyleneglycol-4000 (PEG-4000) and food antigens absorption was found to be similar in the animal intestine. The PEG-4000 gastrointestinal permeability is considerably increased in the rats with anaphylaxis, experimental biliary malabsorption and experimental colitis. Fatty acids of the omega-3 and omega-6 series as well as histidine were found to change the permeability for the PEG 4000. PMID- 8401646 TI - [Proteases of the large intestine]. AB - The large intestine was found to reveal an obvious activity of various aminopeptidases and genuine dipeptidases in dogs and in humans. A dipeptidase was found specific by its independence of the C-terminal configuration of the peptide. This peptidase in an intrinsic membrane protein. Physiological significance of these enzymes is discussed. PMID- 8401647 TI - [The cholinergic mechanisms of the excitatory effects from stimulation of the beta-adrenoreceptors in the pyloric sphincter and small intestine]. AB - Jb been shown that beta-adrenergic stimulation by isadrine can lead to decreasing as well increasing of spre activity of the pyloric sphincter and small intestine administration of isadrine in conditions of the blockade of M- and N cholinoreceptors suppressed the spike activity in all the portions of the intestine. The data obtained suggest that excitatory beta-adrenoreceptors are located on the cholinergic interneurons of the enteral nervous system. The suppression of the spike activity was, evidently, due to a stimulation of postsynaptic beta-adrenoreceptors. PMID- 8401648 TI - [The effect of lactin on histamine-stimulated gastric secretion and choleresis]. AB - Lactine was found to depress the level of the histamine secretion of the gastric juice and to reduce the debit of hydrochloric acid in dogs. This peptide suppressed mainly the spontaneous choleresis and alters the compound composition of the bile. Large doses of the hormone obviously suppress the transport of bilirubin into the secret. PMID- 8401649 TI - [The hormonal regulation of bile secretion due to the activation of protein biosynthesis in the hepatocytes]. AB - The enhanced bile secretion induced by hydrocortisone coincides in time with activation of the biosynthesis of intracellular and export proteins of the liver. Preliminary inhibition of the protein biosynthesis in the hepatocytes hinders the development of hypercholeretic effects of hydrocortisone or considerably distorts them. The activation of the protein biosynthesis in the liver cells is one of the main molecular mechanisms of choleretic action of glycocorticoids and a number of other hormones. PMID- 8401650 TI - [The effect of alimentary factors on the functional development of the forestomach in calves in the early period of ontogeny]. PMID- 8401651 TI - [The current concepts of substance transport across the pre-epithelial layer of the small intestine]. PMID- 8401652 TI - [The mechanisms of the regulation of the assimilation of organic and inorganic compounds]. PMID- 8401653 TI - [The role of dopamine in the mechanism of the regulation of the digestive transport functions of the enterocyte membrane during stress]. AB - The L-DOPA and sulpiridinum effects on the enzymatic transporting function of the enterocytes' membrane, were studied in rats under stress. The L-DOPA was shown to hinder the absorption rate of glucose, whereas sulpiridinum was shown to suppress the main enzymes excepting snerase and alkaline phosphatase, to increase the monomeric and hydrolysate glucose transport. The L-DOPA seems to play a modifying role in the regulation and integration of the digestive-transporting function of the enterocytes' membrane in rats under stress. PMID- 8401654 TI - [An inhibitory analysis of the role of the enterocyte cytoskeleton in the absorption of food substances in the small intestine]. AB - The effect of colchicine and cytochalasin B and D on the process of glucose and plant oil absorption in the small intestine of rats was studied using the light and electron microscopy and biochemical methods. The colchicine and CB, CD action on the elements of enterocytes' apical contractile complex and cytoskeleton inhibited the absorption thus suggesting the major role of endocytosis in the process of nutrients absorption in the small intestine. PMID- 8401655 TI - [An analysis of the structural characteristics of the tight junction of the enterocytes of the rat small intestine during nutrient absorption (immunoelectron microscopic research)]. AB - To analyze structural changes of intercellular relationships of the enterocytes during glycine, glucose, and triolein absorption the structural and immunocytochemical methods of electron microscopy were used. The study was carried out on the proximal part of the rat small intestine in acute and chronic experiments. In the acute ones glucose or glycine solutions (both of 10 and 40 mM) or triolein emulsion (0.5%) were inserted into the isolated small intestinal segments for 20 min. In chronic experiments the isolated loop of the small intestine was perfused with glucose solution (40 mM). Then the corresponding pieces of the intestinal tissue were fixed for structural and immunocytochemical studies. Rarely (in 1% of all cases), and only in chronic experiments, structural changes in the tight junctions--"blisters" and dilatations--were found. At the same time the analysis of the spatial distribution of actin filaments showed that during glucose and glycine absorption the antiactin tracers were found not only within microvilli and on the "root" filaments but also in vicinity of the tight junction and between terminal filaments. The results obtained agree with the hypothesis about the possibility of paracellular transport of some nutrients induced by sodium-dependent transport of glucose and glycine. PMID- 8401656 TI - [The relative role of different mechanisms of glucose absorption in the small intestine under physiological conditions]. AB - Chronic experiments in animals have shown that, even at the maximal glucose (75 mM) and maltose (37.5 mM) concentrations in the initial perfusate, the rate of glucose transfer by a solvent drug does not exceed 10 and 13 per cent, resp. of the total rate of glucose absorption in an isolated intestinal loop. The absorption of glucose under physiological conditions seems to be due mainly to its active transport across the apical membrane of the enterocytes. PMID- 8401657 TI - [The effect of the peptide constituents of casein hydrolysates on glucose and water absorption in the rat small intestine]. AB - The effects of the peptide constituents of two casein hydrolysates applied to the serosal surface, involved the activation of the water absorption alone. Application of these peptides to the mucosal surface slightly inhibited both the glucose and water absorption. The data obtained suggests the mechanism of the peptides action on the glucose and water absorption in the small intestine. PMID- 8401658 TI - [The effect of starvation on carbohydrate transport in the mouse and rat small intestine]. AB - The starvation-related changes of glucose and maltose absorption were shown to be unrelated to absorption of sucrose and lactose. The glucose and maltose transport responds synchronously to starvation and refeeding. The total reduction of the mice body mass after refeeding occurred in 48 hrs. The effect of a preliminary load of the gastrointestinal tract on the transport rate in enzyme-transport system, is discussed. PMID- 8401659 TI - [The physiological mechanisms of the delay in the functional maturation of the small intestine in rat pups prematurely switched from milk to definitive nutrition]. AB - Weaving of 15-day rats delays the natural suppression of the small intestine lactase, induces sucrase activity and decreases the thyroxine level in the blood serum. Administration of thyroxine normalizes the level of this hormone in the blood serum as well the beginning of suppression of the lactase activity. PMID- 8401660 TI - [The effect of nitrates on rats during pregnancy on the development of the digestive system in ontogeny]. AB - Administration of nitrates to pregnant rats induced disorder in maturation and hormonal regulation of enzymes prompting the digestion of the main nutrients in the small intestine of suckling rats. Essential changes were also found under these conditions in activity of the liver and kidney enzymes maintaining the barrier and protective functions. PMID- 8401661 TI - [The intracellular mechanisms of signalling in the parietal cells of the gastric mucosa]. PMID- 8401662 TI - [The effect of hyperparathyroidism on the function of the hypothalamo-hypophyseal adrenocortical system in unilaterally adrenalectomized rats]. AB - Hyperparathyroidism was shown to activate the hypothalamo-hypophyseal adrenocortical system (HHACS) in rats, intensifying the compensatory adrenal hypertrophy in unilaterally adrenalectomised rats, rising the level of 11 oxycorticosteroids in their plasma and adrenal gland, activating the production of corticoliberin in hypothalamus and corticotrophin in hypophysis. The HHACS activity was the highest in 3-month-old rats. PMID- 8401663 TI - [The role of the structural-functional characteristics of the vascular wall in the mechanism of action of erythropoietin on the activity of smooth-muscle cells]. PMID- 8401664 TI - [An automated unit for studying the effect of discontinuous laser radiation on the isolated heart]. PMID- 8401665 TI - [The processes of foresight and of the evaluation of behavioral results and their cellular neurochemical mechanisms]. AB - Chemical sensitivity of the cat visual cortex neurons was studied in conditions of rewarded and unrewarded instrumental behaviour. A connexion was found between the processes of provision and estimation and the neurochemical mechanisms of the central neurons. The unit spike activity was found to depend on the protein synthesis in the neurons. PMID- 8401666 TI - [The views of I. S. Beritashvili on psychoneural activity and the principle of the dominant]. PMID- 8401667 TI - [The form of the spontaneous EEG waves under different conditions of mental activity]. AB - Value of the average level of asymmetry of the EEG waves' fronts (ALA) was shown to decrease in posterior leads and increase in anterior leads during the transition from the rest to mental activity. An inter-hemisphere asymmetry of the ALA was found, the asymmetry having a similar direction for all the pairs of symmetrical leads. Two independent factors are supposed to affect the ALA: the level of the cortex activation and the specifics of information processing by the given hemisphere. PMID- 8401668 TI - [The characteristics of the cholinergic mechanisms of the nucleus accumbens and the caudate nucleus in regulating an instrumental escape reflex in dogs]. AB - We perform comparative analysis of the involvement of dorsal (Neostriatum) and ventral (Nucleus Accumbens) striatal cholinergic systems in the regulation of defensive instrumental reflex, connected with the maintenance of certain flexor posture, in dogs. The data obtained show that the carbachol microinjections (0.05 mkg) into the contralateral Head of Caudate Nucleus significantly improved instrumental reflex due to increasing of a tonic movement component and inhibition of interstimulus and phasic leg raising. In was shown that the cholinergic system of Neostriatum was of great importance for the regulation of the main component of postural adjustment: the unloading of the working limb. The same microinjections into Nucleus Accumbens significantly improved (ipsi- and contralaterally) the instrumental reflex due to prolonged increasing of a phasic component of movement and its amplitude. It can be suggested that the Neostriatum cholinergic system is structurally included into the motor components realization of instrumental reflex, connected with a certain posture maintenance. The activation of cholinergic system of Nucleus Accumbens affects this form of behavior in an excitatory way, being of nonspecific and prolonged character. PMID- 8401669 TI - [The effect of the stimulation of sections of the pons that inhibit movement on the neuronal activity of the medial medulla oblongata]. AB - Out of 249 units of the medial medullary area, 40 were ineffective, 25 fired antidromically, 48--orthodromically, 72 were inhibited and 64 developed a tonic firing pattern during stimulation of inhibitory pontine areas. The data obtained revealed that somatosensory neurons were mostly inhibited whereas reticulospinal neurons suppressed the activity of the hindlimb motoneurons. PMID- 8401670 TI - [The effect of endogenous enkephalins on the phenomenon of the synchronization of vagus and cardiac rhythms]. AB - The effect of met- and leu-enkephalins in cats was characterised by depressing vagotropic action due to a selective decrease in the tonic component. The potentiation of synchronizing actions may serve as an index of a reliable and stable mechanism of the beat-by-beat control of the cardiac rhythm. PMID- 8401671 TI - [The microviscosity of the whole blood in rats with acute hypoxia]. AB - The EPR spectrum was optimally recorded at 20 degrees C in the blood sample. The rotatory ability of the probe, expressing the microviscosity of the blood, was shown to depend on the dose of administered solution of the sodium nitrite and to correlate with the degree of hypoxic state. An analysis of probable causes of the changes in the blood microviscosity in acute hypoxia was carried out. PMID- 8401672 TI - [The activity and properties of K-pNPphosphatase in membrane preparations of erythrocytes with different polypeptide compositions in Wistar, Wistar-Kyoto and SHR strain rats]. AB - A higher K-pNPPase activity was found in erythrocyte membrane preparations from the SHR as compared with the Wistar rats. Removal of peripheral proteins from the membrane skeleton depressed the K-phosphatase activity and eliminated the difference between the SHR and Wistar rats. No activating effect of the ATP and Ca on the enzyme was found. The data obtained suggest that the proteins of the erythrocyte membrane skeleton are connected with regulation of the enzyme transport in the erythrocyte membranes of the SHR via a modulating effect of intracellular ATP and Ca. PMID- 8401673 TI - [Changes in the cardiorespiratory parameters during work at a constant rate of heart contractions]. PMID- 8401674 TI - [The role of adrenoreceptors in the manifestation of the smooth-muscle reactions of the pyloric sphincter and small intestine to acute blood loss]. AB - Acute blood loss evoked an increase in the spike activity of the pyloric sphincter' smooth muscles, those of the duodenum and ileum. No such effect was found in joint blockade of alpha- and beta-adrenoreceptors. The increase in the spike activity seems to be due to an effect of endogenous catecholamines upon activating alpha- and beta-adrenoreceptors of the pyloric sphincter, duodenum and ileum. PMID- 8401675 TI - [The effect of changes in the blood flow and the level of arterial and venous pressures on the filtration-absorptive processes in the small intestine]. AB - Under conditions of perfusion of the small intestine's vessels with constant volume of the auto-blood, stationary shifts of the arterial perfusion pressure by +/- 50 mm Hg and/or blood flow by +/- 50% of the initial level practically did not affect the size of the exchange surface of the organ's microvessel bed (the capillary filtration coefficient) or the average capillary pressure in artificially stabilised pressure of venous outflow at the level of 6 mm Hg. However, under conditions of normo-, hypo- and hypertone of the small vessels' smooth muscles, the shifts of the venous outflow pressure (from 0 to 24 mm Hg) increased the exchange surface of the microvessels and the average capillary pressure. The mechanisms of the shifts in microhemodynamics and transcapillary exchange of fluid are discussed. PMID- 8401676 TI - [A comparative analysis of the feedback inhibition of pancreatic secretion from the duodenum in acute and chronic experiments]. PMID- 8401677 TI - [The development of interrelationships between the activity-rest cycle and thermoregulation in rabbit fetuses]. AB - The gradient of temperatures of the foetus and mother was found to increase along with an increase of the gestation age which coincides with an increase in the mass of grey fat tissue. In artificial placenta insufficiency, the activity-rest cycle of the foetuses is delayed by 1 or 2 days in their development. The mechanisms of thermoregulation were shown to start forming by 4 or 5 days earlier than the activity-rest cycle, the character of their functioning being comparable to that in poikilothermic animals. PMID- 8401678 TI - Origins and effects of variations in spermatozoal quality. AB - A traditional view of mammalian fertilization is that the active component of the process, the spermatozoon, by virtue of its progressive motility and acrosomal enzymes, penetrates an otherwise passive oocyte. This concept has placed bias on spermatozoal normality as largely determining the outcome of fertilization. Once this has been achieved, the contribution of the spermatozoon is often forgotten, and attention switches to the maternally derived "blue-print" for early embryonic development. Paternal genomic contribution is known to start at the 8-cell stage in the human, but this is usually after the early cleavage stage embryos are transferred in human reproductive technologies, such as in vitro fertilization (IVF). Hence, any fundamental abnormal contribution to embryogenesis derived from the fertilizing spermatozoon is not seen. IVF has permitted far greater powers of analysis of fertilization in the human, and fertilization success appears to be determined in this system by three main factors: spermatozoal quality, oocyte quality, and the quality of the in vitro culture conditions (the gamete environment). If the second two factors are more carefully controlled than the first, as is the usual emphasis in IVF practice, then any large variation in fertilization rates that is also related to embryonic viability and ultimate pregnancy outcome may be thought to be more directly associated with original quality of the fertilizing spermatozoon. If this hypothesis is accepted, we should drastically alter our concept of the spermatozoon as a robust simple initiator of embryonic development, and embrace the idea of the vulnerability of such germ cells both during and after their production, and how detrimental influences on this may profoundly affect embryogenesis after fertilization. PMID- 8401679 TI - A comparative multicenter study of two transdermal estradiol replacement therapies in the treatment of postmenopausal symptoms. AB - OBJECTIVE: Comparison of the effects of treatment of two transdermal therapeutic systems for estrogen replacement therapy with regard to efficacy, tolerability, and acceptance. DESIGN: Open randomized. SETTING: Multicenter. PATIENTS AND INTERVENTIONS: A study population of 104 postmenopausal women was randomized on a 1:1 basis to treatment with one of two estradiol patches, System (Cilag) and Estraderm (Ciba-Geigy). OUTCOME MEASURES: Systolic and diastolic BP, hot flushes, night sweating, fatigue, insomnia, depression, nervousness, headache, vaginal discomfort (efficacy variables); bleeding, dermatological symptoms, comfort and adhesiveness of patch, and other possible causes of discontinuation (tolerability); general evaluation by patient (acceptance). RESULTS: Considering all efficacy variables, 53% of Systen and 46% of Estraderm patients found the therapy satisfactory. Tolerability was somewhat higher in the Systen group. Adhesiveness of the patch was significantly better for Systen. Overall, 79% of Systen patients and 62% of Estraderm patients evaluated treatment as "good" or "very good." The majority of patients in both groups found the patch very comfortable or only slightly obtrusive. PMID- 8401680 TI - HLA frequency, HLA sharing and immunotherapy in the management of recurrent miscarriage. AB - OBJECTIVE: To test couples with three or more consecutive recurrent miscarriages for suitability of the mother for immunization with paternal lymphocytes. SETTING: Regional transfusion centre, patients referred by local obstetricians. PATIENTS AND INTERVENTIONS: Women treated were from 103 couples with > or = 3 consecutive recurrent miscarriages and no more than one live birth. HLA sharing and frequencies were investigated, and contrasted with control data. Patients showed increased sharing versus control (but not in comparison with gene frequencies of the county population). Sixty-eight mothers were finally found suitable and immunized with paternal lymphocytes. OUTCOME MEASURES: Pregnancy and live births. RESULTS: 55/68 conceived, with 30 live births resulting; 8/13 additional successes after repeat (booster) immunization. CONCLUSIONS: This treatment still suffers from the lack of clear criteria for patient selection, and it should be submitted to a prospective controlled double-blind trial. PMID- 8401681 TI - Peripheral catecholamine metabolites and menstrual irregularity in patients with polycystic ovaries. AB - OBJECTIVE: To investigate the catecholamine status of patients with polycystic ovary syndrome. DESIGN: Three parallel groups with polycystic ovary were diagnosed by ultrasound: (a) 5 patients with regularly ovulatory menstruation; (b) 10 with anovulatory menstruation; (c) 13 with secondary amenorrhea who responded to progestagen with withdrawal bleeding. Blood samples for measurement of LH, testosterone, and catecholamine metabolites were drawn during cycle days 4 7. RESULTS: (1) Serum LH and testosterone of the patient groups (b) and (c) were significantly higher than those of controls. (2) Plasma 3,4-dihydroxyphenylglycol (DOPEG) and the DOPEG/DOPAC ratio were elevated in patients, and DOPAC levels were reduced. However, there was no significant difference of catecholamine metabolites among the three patient groups. CONCLUSIONS: The androgen status in polycystic ovary diagnosed by ultrasound is correlated, but catecholamine status is not correlated, with the menstrual irregularity of polycystic ovary syndrome. PMID- 8401682 TI - Leukocyte subpopulations and natural killer activity in endometriosis. AB - OBJECTIVE: To evaluate cell-mediated immunity in endometriosis. DESIGN: Peritoneal fluid and peripheral blood samples from 19 women with macroscopic evidence of endometriosis and 26 controls were submitted to 2-color flow cytometric analysis to assess lymphocyte subsets, using various combinations of monoclonal antibodies. K562 cells were used in a routine assay by 51Cr release to measure the cytotoxicity of NK cells in the subjects' peripheral blood. RESULTS: Suppressor T-cells were significantly increased and cytotoxic T-cells significantly decreased in both peritoneal fluid and peripheral blood of the women with endometriosis. NK activity in peripheral blood was decreased in endometriosis patients as compared with controls. CONCLUSION: An alteration in cell-mediated immunity may be among the pathogenetic, or developing, factors in endometriosis. PMID- 8401683 TI - In vitro fertilization and embryonic development of oocytes fertilized by sperm treated with 2-deoxyadenosine. AB - OBJECTIVE: To test the effect in an assisted pregnancy program of an agent known to enhance sperm motility and oocyte penetration ability. DESIGN: Prospective with internal control. SETTING: Hospital clinic. PATIENTS AND INTERVENTIONS: Forty-two oocytes obtained from women undergoing IVF were inseminated with their husband's sperm washed, incubated, and capacitated in Earle's medium supplemented with 1 mM, 2.5 mM, or 5 mM 2-deoxyadenosine (2-DXA). The outcome of insemination was compared with those of 234 control oocytes from the same women that were inseminated with husband's sperm washed in 2-DXA-free medium. 15, 14, and 13 oocytes were inseminated with sperm washed in 1 mM, 2.5 mM, and 5 mM 2-DXA, respectively. RESULTS: Fertilization rates in the respective groups were 93%, 85.7%, and 85.5%. These were higher than the 70.1% fertilization rate in the control group, but only statistically higher (P < .002) in the 1 mM 2-DXA group. Embryonic development in all three 2-DXA groups was comparable to the controls. CONCLUSION: It is suggested that the value of sperm motility enhancing agents requires further evaluation in assisted pregnancy programs. PMID- 8401684 TI - Can staging systems for endometriosis based on anatomic location and lesion type predict pregnancy rates? AB - OBJECTIVE: Development of an endometriosis classification system based on empirically derived stages of the disease, to supplant the Acosta (1973), Kistner (1977), and American Fertility Society (1985) classifications, which are based on arbitrarily defined stages and often fail to predict pregnancy rates. DESIGN: Retrospective cohort analysis. SETTING: University infertility clinic. PATIENTS AND METHODS: Women with endometriosis and > or = 1 year of infertility. Diagnosis of endometriosis was made by direct visualization, with type of lesion (implant or adhesion) at multiple sites recorded; total of 202 patients. All diagnosed infertility problems were treated based on semen analysis, postcoital test, and endometrial biopsy. Pregnancy rates were analyzed by life-table and cluster analyses, and combinations of site and type were also analyzed by Cox's regression model. RESULTS: No individual anatomic site or type significantly affected prognosis, nor was any cluster useful for predicting outcome. CONCLUSION: Anatomic site and type of lesion are insufficient for predicting fertility when used as sole components of a clinical staging system for endometriosis. PMID- 8401685 TI - Left adrenalectomy in varicocelized rats does not inhibit the development of varicocele-related physiologic alterations. AB - OBJECTIVE: To evaluate the importance of retrograde adrenal metabolite flow for testicular function in a varicocelized animal model. MATERIALS AND METHODS: Partial occlusion of the left renal vein and ligation of the collateral vein between the left testicular and left common iliac veins was carried out on 14 Wistar rats (group A); same procedure plus adrenalectomy on 14 others (group B); seven sham operated (C) and seven with adrenalectomy alone (D). RESULTS: Groups A and B showed significant reduction in fertility, and also bilaterally in difference between testicular and abdominal temperatures, epididymal spermatozoal content and motility, and testicular weight at 12 weeks post-surgery. All values were nearly identical in groups A and B. CONCLUSIONS: Since left adrenalectomy did not ameliorate the effects of varicocele, it appears that retrograde flow of adrenal metabolite(s) may not be responsible for the harmful effect of (left) varicocele on fertility. PMID- 8401686 TI - Reform praised, but stakeholders concerned with implementation. PMID- 8401687 TI - Should organ transplant centers be federally designated and controlled? No. PMID- 8401688 TI - Should organ transplant centers be federally designated and controlled? Yes. PMID- 8401689 TI - CEO poll. An elusive balance: operations and general skills. PMID- 8401690 TI - Where ex-health care professionals go when they leave health care. PMID- 8401691 TI - Diagnostic imaging procedures show increase. PMID- 8401692 TI - Quantum leaps. A look at the health care delivery system in twenty years. AB - What will the health care delivery system of the future look like? Though 20 years from now may seem like a long way off for some people, health care experts are already seeing the seeds of future realities begin to sprout. But the new era of delivery also will involve many features now just barely imaginable, including wearable PCs, gene therapy, a diffusion of hospital services across a broad geographical area, and computer-based diagnoses. PMID- 8401693 TI - Universal access? Undocumented residents and reform: how will we pay for their care? PMID- 8401694 TI - Reinventing the system. The re-engineering process will launch a new era of health care delivery. PMID- 8401695 TI - Back to school. Market demands are compelling some specialists to seek retraining. PMID- 8401696 TI - Wausau, Wisconsin. Hospital seeks partnerships to develop regional health network. PMID- 8401697 TI - Data watch. Future plans--anxious about the future, hospitals step up planning. PMID- 8401698 TI - AHA adopts new structure for strategic planning. PMID- 8401699 TI - A new MBA program focuses on practice management. PMID- 8401700 TI - Hospitals improve balance sheets in preparation for reform. PMID- 8401701 TI - Supply meets demand: hospital to tie critical paths, purchasing. PMID- 8401702 TI - Rural hospitals set their sights beyond acute care. PMID- 8401703 TI - Building the data intensive hospital. PMID- 8401705 TI - An endpoint that marks a new beginning. PMID- 8401704 TI - Three's company. Hospital, physicians and community make a strong alliance. Interview by Therese Hudson. AB - One 250-bed hospital in rural Pennsylvania illustrates the old adage "necessity is the mother of invention." It recently found that health care delivery had to change, so it created a PHCO--a physician, hospital, community organization that directly contracts with employers for health care. This was an innovative move for Meadville Medical Center, a sole community provider serving 80,000 in Crawford County. But the innovation doesn't stop there. The PHCO is moving toward capitation, says Meadville CEO Anthony J. DeFail. One day, he'd like to see all the hospital's business come through the PHCO. DeFail talked to senior editor Terese Hudson about the PHCO's origins. PMID- 8401706 TI - Groups position themselves for the health care reform debate. Making moves. PMID- 8401707 TI - The FDA's list of devices tracked as of Aug. 29. PMID- 8401708 TI - Mapping care. AB - Managing clinical quality and assessing the efficacy of clinical operations are key objectives for all health care providers--but pressure is growing to do more with fewer resources. As a result, many innovators are turning to critical paths as a tool that can help them achieve those goals. Indeed, a new survey by Hospitals & Health Networks and Medicus Systems Corp. finds critical paths are taking hold in health care organizations in a big way. Of those hospital executives surveyed, 57 percent say they have a critical paths program. But the survey also uncovered a slew of names for the tools being used, and experts agree there are wide disparities in innovators' motivations, methods and achievements. PMID- 8401709 TI - Dinosaur or chameleon? PPOs face a future in which survival demands risk-sharing arrangements. PMID- 8401710 TI - San Francisco. Reconfiguring the system takes different forms. PMID- 8401711 TI - Letting telemedicine do the walking. AB - Telemedicine is becoming a major tool for bringing clinical expertise to rural areas. The cost savings could be substantial once regulatory, reimbursement and infrastructure obstacles are overcome. PMID- 8401712 TI - Data watch. Network sizing. Utilization patterns differ by payer, region and industry. PMID- 8401713 TI - Breaking the bank? MDs say NPDB needs screening. PMID- 8401714 TI - 'Stark II' limits physicians, but may help hospitals. PMID- 8401715 TI - Need vs. want. Will consumers decide? PMID- 8401716 TI - The inspiration of Stella Mpanda. PMID- 8401717 TI - Inverted uterus. PMID- 8401718 TI - Who has influenced you most in your birth practice? PMID- 8401719 TI - Birth in Holland. Interview by Kim James. PMID- 8401720 TI - Chemical dependency in pregnancy. PMID- 8401721 TI - Homebirth "underground". PMID- 8401722 TI - Healing through birth. PMID- 8401723 TI - How cerebral palsy affected my births. PMID- 8401724 TI - The garden room. PMID- 8401725 TI - Guidelines for serving disabled women. PMID- 8401726 TI - Empowering midwifery through unity. PMID- 8401727 TI - Panic attacks and pregnancy. PMID- 8401728 TI - An awkward step. PMID- 8401729 TI - Dystocia dialogue. PMID- 8401730 TI - Change is brewing. PMID- 8401731 TI - Time to counsel. PMID- 8401733 TI - Be clear about money. PMID- 8401732 TI - Legislation unwise. PMID- 8401734 TI - Action against harassment. PMID- 8401735 TI - Ancient disease on increase. PMID- 8401736 TI - Nurses debate similar issues. PMID- 8401737 TI - Making sense of health policies. PMID- 8401738 TI - Plunket must survive. PMID- 8401739 TI - Preventing childhood accidents. PMID- 8401740 TI - A range of partnerships. PMID- 8401741 TI - Looking back with pride. PMID- 8401742 TI - Suffrage. A pioneer for nursing. PMID- 8401743 TI - Anaesthesia and the obese patient. AB - The obese patient is likely to have pre-operative impairment of cardiovascular and respiratory function. These impairments will tend to increase in the per- and post-operative period and place the patient at risk of myocardial ischaemia and hypoxaemia. The physical effects of obesity mean that all patients require endotracheal intubation during anaesthesia and this may be difficult to achieve. The difficulties in moving and positioning the patient and difficulties in gaining access for monitoring and venous cannulation add to the problems. Anaesthesia and surgery may present a considerable risk for obese patients and should not be undertaken without full understanding of the potential problems. PMID- 8401744 TI - Relationships of lipid and glucose metabolism with waist-hip ratio and physical fitness in obese men. AB - The waist-hip ratio (WHR) is an indirect index of abdominal type obesity which has been shown to be strongly correlated with the risk of coronary heart disease. Empirically, men who have a higher WHR seem to have a lower level of physical fitness. In the present study, the relationships of lipid and glucose metabolism with WHR and physical fitness were examined in 207 Japanese obese men. Physical fitness was evaluated by the oxygen uptake at the onset of blood lactate accumulation (OBLA-VO2; ml/kg/min) which corresponds to 4 mmol/l of blood lactate during graded exercise test and is one of the best indicators of the muscle oxidative capacity. The WHR and percentage of body fat (% body fat) were significantly correlated with OBLA-VO2. The WHR and % body fat were significantly related to each other. After adjusting for % body fat, a significant negative correlation was observed between the WHR and OBLA-VO2 (r = -0.24; P < 0.05). A multiple linear regression was calculated for parameters relating to lipids, and glucose and insulin areas separately, when the WHR, OBLA-VO2, % body fat and age were entered as independent variables. OBLA-VO2 significantly showed a negative relationship with triglyceride (TG), HDL-C/total cholesterol, and the insulin area, while the WHR was only independently related to TG. From these results, we conclude that the physical fitness level evaluated by OBLA-VO2, which represents the capacity of the aerobic metabolism in muscle, may thus be a determinant of lipid and glucose metabolism in obese men. PMID- 8401745 TI - A comparison of adiposity measures for screening non-insulin dependent diabetes mellitus. AB - We compared the accuracies of adiposity distribution measures (waist and hip circumferences, subscapular and triceps skinfold thicknesses, waist-hip and subscapular-triceps ratios) with a measure of adiposity extent (body mass index) for screening non-insulin dependent diabetes mellitus (NIDDM). Of 521 subjects (218 men and 303 women) who had 2 h oral glucose tolerance tests, 43 men and 28 women were found to have NIDDM. Allowing for age, ethnicity, and family history in analyses of covariance, the subscapular skinfold thickness and the ratio of the subscapular to triceps skinfold thicknesses were significantly higher (P < 0.05) in both the men and women found to have diabetes than in their non-diabetic counterparts. The waist-hip ratio was also significantly higher in diabetic women; however, the elevation was not as marked in diabetic men (P = 0.06). The body mass index did not differ between diabetic and non-diabetic men and was only moderately elevated in diabetic women (P = 0.04). Receiver-operator curves were employed to examine the relative accuracies of the body mass index, waist-hip ratio, and subscapular skinfold thickness for screening NIDDM. In both men and women, the waist-hip ratio and subscapular thickness were superior to the body mass index. These data suggest that certain measures of adiposity distribution are more accurate than measures of overall adiposity extent for screening NIDDM and that they may be useful in screening programmes. PMID- 8401746 TI - Decreased physical activity in Pima Indian compared with Caucasian children. AB - Since reduced physical activity might be a risk factor for body weight gain, we studied the relationship between physical activity and body composition in 43 Pima Indian children (22 male/21 female, mean +/- s.d.: 9.9 +/- 1.1 years) and 42 Caucasian children (21 male/21 female, 9.7 +/- 1.2 years). A list of usual sport leisure activities was established (e.g. bicycling, swimming, basketball) and the subjects were asked how much time they had devoted to each activity over the past week and the last year. Data on time spent playing outside (excluding sport leisure activities for the estimation of physical activity) and watching television/videos were also collected. Pima Indians were taller (143 +/- 9 vs. 137 +/- 8 cm, P < 0.001), heavier (48.6 +/- 15.8 vs. 32.9 +/- 7.8 kg, P < 0.0001) and fatter (39 +/- 16 vs. 24 +/- 7% fat, P < 0.001) than Caucasians. Pima Indian girls showed significantly lower past year and past week sport leisure activity than Caucasian girls (P < 0.01) and spent significantly more time watching television/videos (P < 0.05). Pima boys also showed significantly lower past week sport leisure activity than Caucasian boys (P < 0.05). In Pima Indian boys, past year sport leisure activity correlated negatively (P < 0.05) with body mass index (r = -0.49) and percentage body fat (r = -0.56). However, such correlations were not found in Pima Indian girls, possibly due their very low levels of activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8401747 TI - Gastric banding for morbid obesity: five years follow-up. AB - This paper reports on a five year follow-up of 174 morbidly obese patients (132 women) with gastric banding performed between 1981 and 1985. Mean preoperative weight was 122.6 +/- 1.4 kg (s.e.m.) (body mass index (BMI) = 41.8 +/- 0.4 kg/m2) and mean overweight was 73.2 +/- 1.6%. After rapid weight loss during the first six months weight levelled off reaching a nadir at 12-18 months. At 12 months mean weight loss was 36.5 +/- 1.2 kg (BMI = 29.1 +/- 0.4 kg/m2). At 60 months BMI had increased to 32.3 +/- 0.6 kg/m2 (P < 0.05 vs. 12 and 24 months). Mean excess weight at 60 months was 33.5 +/- 2.4% with 47.5% of patients maintaining less than 30% overweight. There were no differences in relative weight loss between men and women and no differences between stomal diameter of 12 or 15 mm 60 months after the operation. Early post-operative complications occurred in 25 patients (14.4%), four of whom required reoperation. Three of these re-operations were for perforations, one of which was fatal. Forty-eight patients (28%) had altogether 60 late complications requiring 26 reoperations (14.9%). There has been a total of four deaths (one clearly unrelated, one unknown) in the series. We conclude that gastric banding is a simple and safe gastric restrictive operation that is effective in about 50% of patients weighing between 90-181 kg (BMI 33-69 kg/m2). PMID- 8401748 TI - Relationships and independence of body composition, sex hormones, fat distribution and other cardiovascular risk factors in overweight postmenopausal women. AB - The relationships and independence of body composition, sex hormones, fat distribution and other cardiovascular risk factors were studied in 121 overweight postmenopausal women. Body composition and fat distribution were directly measured by dual energy X-ray absorptiometry (DXA); other parameters were waist to-hip ratio (WTH), sex hormone binding globulin (SHBG), testosterone, androstenedione (A), oestradiol (E2), and lipids and lipoproteins; cigarette and alcohol consumption were recorded. SHBG was correlated with the fat distribution by DXA and WTH (r = -0.35, P < 0.01) and A with the fat mass (r = -0.3, P < 0.01). SHBG had a negative and alcohol and cigarette consumption a positive, independent relationship to a central fat distribution (by DXA and WTH, respectively) (P < 0.01). SHBG (r = 0.23, P < 0.05), fat distribution (WTH) and A (r = -0.2 to -0.3, P < 0.01) were correlated with high density lipoprotein cholesterol (HDL-C). However, only fat distribution (WTH) and A were independently related to HDL-C (P < 0.05). Fat distribution (WTH) and SHBG were independently related to triglycerides (P < 0.05), whereas fat distribution (by DXA) and E2 were independently associated with total cholesterol and low density lipoprotein cholesterol (LDL-C) (P < 0.05). Thus, in overweight postmenopausal women, androgenicity and cigarette and alcohol consumption were independently positively related to a central fat distribution. Furthermore, atherogenic levels of lipids and lipoproteins were independently related to a central fat distribution, androgenicity and low levels of oestrogens. PMID- 8401749 TI - Sex differences in catecholamine response to clonidine. AB - Sex-related differences both in the basal secretion of catecholamines and in the adrenergic reactivity to various stimuli have been described. We studied the responses of catecholamines and arterial blood pressure to clonidine (0.3 mg per os) in 31 normotensive subjects (10 men (M), aged 18-42 years, and 21 women (W), aged 20-48 years). Plasma catecholamines were determined by HPLC at -30, -15, 0, 120, 130, 140 min after clonidine. The basal levels of plasma norepinephrine were similar in men and in women (M = 1.16 +/- 0.26 vs. W = 0.87 +/- 0.07 nmol/l). Basal plasma epinephrine levels were not different in the two sexes (M = 0.21 +/- 0.03 vs. W = 0.14 +/- 0.03 nmol/l). The mean arterial pressure decrease after clonidine was similar in the two groups (M = 13 +/- 3 vs. W = 15 +/- 2 mmHg). The decrease in plasma epinephrine after clonidine was similar in men and women (M = 0.06 +/- 0.04 vs. W = 0.09 +/- 0.02 nmol/l). In contrast, the plasma levels of norepinephrine after clonidine were reduced more in women than in men either when expressed as absolute values (W = 0.63 +/- 0.07 vs. M = 0.3 +/- 0.1 nmol/l; F = 7.6, P < 0.02) or as percentage change (W = 71 +/- 3 vs. M = 34 +/- 8; P < 0.002). The present study demonstrates that an elevated alpha 2-adrenergic activity in women may be responsible for the sexual dimorphism in catecholamine secretion. PMID- 8401750 TI - Weight-for-height of south Iranian school children and the NCHS reference data. AB - Weight-for-height charts for pre-pubescent school boys (aged 6-11 years) and girls (aged 6-10) years in Shiraz, Iran, and their relationships to the NCHS data were investigated. Weight-for-height charts for Iranian school children were similar to the NCHS charts except for the higher centiles. Weight-for-height appears to satisfy the need for a simple method to assess nutritional status among school children. The necessity for local weight-for-height reference data for clinical work is emphasized. The weight-for-height charts presented here are likely to be appropriate to all urban school children in Iran. PMID- 8401751 TI - Reliability of B-mode ultrasonic measurements of subcutaneous adipose tissue and intra-abdominal depth: comparisons with skinfold thicknesses. AB - Adipose tissue deposits, particularly intra-abdominal adipose tissue, are associated with health risks such as diabetes mellitus and cardiovascular disease. Anthropometric techniques currently in use can measure subcutaneous adipose tissue (SAT) with skinfold calipers, but are limited to certain sites and cannot measure intra-abdominal adipose tissue (IAAT). Radiography, computerized tomography (CT) and magnetic resonance imaging (MRI) have been used, but they are expensive and some involve exposure to radiation. This study, which investigated the utility of B-mode ultrasound for measuring adipose tissue, found that ultrasonic measurements of SAT were as reliable as skinfold caliper measurements. Intra-observer and inter-observer coefficients of reliability for five of six ultrasonic measurements of SAT ranged from 91-98%, in comparison with coefficients of reliability ranging from 93-98% for three skinfold measurements. Coefficients of reliability for ultrasonic measurements of SAT at the paraspinal site were below 90%. Ultrasonic measurements of intra-abdominal depth (IAD), an index of IAAT, yielded an inter-observer coefficient of reliability of 64%. Ultrasound is recommended for measurement of subcutaneous adipose tissue but not for measurement of IAD. PMID- 8401752 TI - Insulin-like growth factor-1 (IGF-1) and dehydroepiandrosterone sulphate in obese women. AB - Severe obesity is known to reduce either dehydroepiandrosterone circulating levels or growth hormone (GH) and insulin-like growth factor-1 (IGF-1) secretion. The present study was undertaken to evaluate the possibility of a relationship between the circulating levels of IGF-1 and those of dehydroepiandrosterone sulphate (DHEAS) in 25 fertile obese women. A logarithmic transformation of the values of non-normally distributed variables was performed before statistical analysis. We found a significant positive correlation between DHEAS and IGF-1 (r = 0.615, P < 0.01). In addition, stepwise multiple regression analysis showed that IGF-1 maintained a strong positive relationship with DHEAS (P < 0.01) when adjusted for other variables such as age, body mass index (BMI), waist:hip ratio (WHR) and insulin levels (adjusted R2 = 0.373; P < 0.01). These findings suggest that IGF-1 may independently influence the DHEAS circulating levels. ADG (5 alpha androstan-3 alpha, 17 beta-diol-glucuronide) was also positively correlated to IGF-1 (r = 0.436, P < 0.05). However, when ADG concentrations were adjusted for DHEAS levels, this metabolite was not significantly correlated with IGF-1, thus excluding a direct influence of IGF-1 on the 5-alpha-reductase activity. Therefore, although our data represent only a preliminary study, they seem to suggest a possible influence of IGF-1 on circulating levels of DHEAS in obese women. PMID- 8401753 TI - Depressive symptoms and body weight. PMID- 8401754 TI - Agonists and antagonists of adenosine receptors and morphine withdrawal syndrome in rats. AB - The compounds activating adenosine system alleviated morphine withdrawal syndrome (the number of escape attempts and body shakes) in rats. Body shakes were decreased mostly by N-ethylcarboxamidoadenosine, cyclohexyladenosine, dipyridamole and 2-chloroadenosine, while R-phenylisopropyladenosine very strongly decreased the number of escape attempts. Adenosine receptor antagonists (caffeine and theophylline) increased the number of escape attempts in morphine dependent rats. Caffeine (10 mg/kg), administered preventively, antagonized the inhibitory effects of adenosine analogs. These results suggest the existence of relation between opioid receptors and the adenosine system. PMID- 8401755 TI - The effect of vasopressin analog: [d(CH2)(1)5,Tyr(Me)2,delta 3Pro7]AVP on learning and memory processes in rats with experimental amnesia. AB - We investigated the effect of a single 2 micrograms dose of a vasopressin (AVP) analog, [d(CH2)(1)5,Tyr(Me)2,delta 3Pro7]AVP on processes of retrieval, consolidation and acquisition of conditioned reflexes in rats with experimentally induced amnesia. The investigated amnesia models were: long term ethanol intoxication, electroconvulsive shocks (ECS), and hypoxia. They all profoundly impaired the learning and memory processes in all tests used. The AVP analog [d(CH2)(1)5,Tyr(Me)2,delta 3Pro7]AVP facilitated retrieval of passive avoidance in all amnesia models. It improved consolidation of active avoidance of rats previously treated with alcohol, but did not affect the acquisition of active avoidance. [d(CH2)(1)5,Tyr(Me)2,delta 3Pro7]AVP lack antidiuretic properties. PMID- 8401756 TI - The effect of angiotensin II and its fragments on post-alcohol impairment of learning and memory. AB - Angiotensin II(1-8) (A II) and its fragments: angiotensin III(2-8) (A III), angiotensin IV(3-8) (AIV), angiotensin V (4-8) (A V) and angiotensin VI(3-7) (A VI) accelerate acquisition of avoidance response and prolong their extinction. A II fragments are devoid classical A II activities such as the effects on blood pressure and thirst. Alcohol administered chronically (for 9 weeks) depresses the ability to retrieve and acquire avoidance responses. The investigated A II fragments counteract the post-alcohol impairment of learning and memory processes (A V being somewhat less active). Fragments A IV and A VI normalize the retrieval in offspring of mothers exposed to alcohol pre- and post-natally. PMID- 8401757 TI - Psychotropic effects of angiotensin II N-terminal fragments: Asp-Arg-Val-Tyr-Ile His and Arg-Val-Tyr-Ile-His in rats. AB - In this study the effects of angiotensin II (AII) angiotensin II hexapeptide [AII(1-6)] and angiotensin II pentapeptide [AII(2-6)] on the motility, stereotypy, learning of conditioned avoidance responses (CARs) and recall of a passive behavior making it possible to avoid aversive stimulation in rats, were compared. All the peptides were injected into the lateral cerebral ventricle (icv) in a dose of 1 nmol. AII caused a statistically significant increase in the number of crossings, rearings, and bar approaches in an open field whereas [AII(1 6)] and [AII(2-6)] were inactive in this test. The stereotypic behavior induced by an intraperitoneal (ip) injection of apomorphine (1 mg/kg) and amphetamine (7.5 mg/kg) was statistically significantly enhanced only in the rats which received AII icv. The application of AII, but not that of [AII(1-6)] and [AII(2 6)] resulted in a quicker acquisition of the CARs. A better recall of passive avoidance was achieved only by AII, while the fragments [AII(1-6)] and [AII(2-6)] had no effect. These findings indicate that the 1-6 and 2-6 fragments of AII do not possess a psychotropic activity like that of the parent octapeptide. PMID- 8401758 TI - Effect of GABA-ergic drugs on flunarizine-induced seizures in rabbits. AB - In this study the authors have evaluated the effects of common antiepileptic drugs given alone or in combinations with baclofen on seizures evoked by intravenous flunarizine in rabbits. The abolition of clonic and clonic-tonic convulsions was taken as the end point. Baclofen was able to inhibit convulsions only in a relatively high dose (5 mg/kg). Diazepam (1.5 mg/kg) and clonazepam (2 mg/kg) completely abolished clonic seizures. Valproate (20 mg/kg) inhibited clonic seizures and remained without effect on tonic convulsions. Other antiepileptics such as carbamazepine and acetazolamide were found inactive in this respect. Moreover, combinations of acetazolamide (100 mg/kg) and carbamazepine (50 mg/kg) with baclofen (2 mg/kg) had no anticonvulsant effect. Thus, mainly GABA modulatory drugs reveal significant action against flunarizine induced seizures. PMID- 8401759 TI - Nitric oxide donors as generators and scavengers of superoxide anions. AB - Three classical direct nitric oxide (NO) donors, 3-morpholine-sydnonimine (SIN 1), S-nitroso-N-acetyl-d,l-penicillamine (SNAP) and sodium nitroprusside (NaNP) as well as two indirect NO donors, molsidomine (MSL) and glyceryl trinitrate (GTN) were studied for their potencies to generate O2-, to scavenge O2-, to consume molecular oxygen and to inhibit lipid oxidation. Out of five NO donors only those which were spontaneous releasers of NO at physiological pH were also scavengers of O2- which has been generated by xanthine:xanthine oxidase system (SIN-1 IC50 19 microM, SNAP IC50 416 microM) and inhibitors of the Fe3+ and ascorbate stimulated oxidation of rat liver lipids (SIN-1 IC50 76 microM, SNAP IC50 12 microM). Only SIN-1 at high concentrations of 300-5000 microM generated O2- as detected by a SOD inhibitable reduction of nitroblue tetrazolium. None of the in vitro studied activities were exerted by NaNP, MLS and GTN. PMID- 8401760 TI - Influence of acetaldehyde on some serotonergic mechanisms in rat blood platelets. AB - Acetaldehyde (ACT), both, ex vivo and in vitro did not change the ADP-induced rat platelet aggregation and the potentiating action of serotonin. The whole blood serotonin content was decreased only when ACT was used in doses of 20 and 30 mg/kg, i.v. and the platelet serotonin content remained unchanged. Ex vivo, ACT had no influence on the labeled serotonin uptake, whereas in experiments in vitro it inhibited the amine uptake and augmented the serotonin release from blood platelets in a dose dependent manner. The results indicate that all the changes in the platelet serotonergic mechanisms appear only after high concentrations of ACT. They may be of significant importance in the circulatory system or hemostasis only during disulfiram or calcium carbamide therapy. PMID- 8401761 TI - Interference of psychotropic drugs with the injuring action of ethanol on gastric mucosal barrier. AB - The aim of this study was to establish whether some psychotropic drugs, applied to patients under influence of alcohol, may potentiate its damaging action on gastric mucosal barrier. A sensitive index of such damage is a decline of the potential difference (PD) across the stomach wall. The experiments were carried out on Wistar rats of either sex, anesthetized with urethane-chloralose. The PD values were assayed with an apolarization method. The investigated solutions were administered intragastrically by gavage. Ethanol at a concentration of 40% v/v depressed PD by 39%. The investigated psychotropic drugs did not change PD by themselves but given in combination with ethanol caused significant decline of PD: diazepam (0.4 mg/kg) by 58%, chlorpromazine (6.7 mg/kg) by 59%, imipramine (2 mg/kg) by 48%, amitriptyline (4 mg/kg) by 49%, phenytoin (4 mg/kg) by 53%, pyridinol (0.3 mg/kg) by 58%. Intragastric administration of water did not change PD. The results indicate that while psychotropic drugs given alone do not affect significantly the gastric mucosal barrier, they may potentiate the damaging action of ethanol on this barrier. PMID- 8401762 TI - Renin inhibitors containing statine and 6-aminohexanoic acid. Part III. AB - Five peptide renin inhibitors containing the sequence: Phe-His-Sta-epsilon Ahx (Sta = 4(S)-amino-3(S)-hydroxy-6-methylheptanoic acid, epsilon Ahx = 6 aminohexanoic acid) were synthesized and their potency was assayed in vitro by a spectrofluorometric method (assay of Leu-Val-Tyr-Ser released from N acetyltetradecapeptide substrate by renin in the presence of an inhibitor). Their stability was tested by assay of Phe and Pro-Phe released after incubation with chymotrypsin. The most potent inhibitor was Boc-Phe-His-Sta-epsilon Ahx-OMe (IC50 = 5 x 10(-9) M/l), the most stable--Boc-Pro-Phe-His-Sta-epsilon Ahx-OMe (resistant to incubation with chymotrypsin for 4 h). PMID- 8401763 TI - TRH analogs with 4-nitro- and 4-N-butyloxycarbonylamino-1-methyl-2 pyrolylcarboxylic acid in position 1. Synthesis and biological properties. AB - Two new analogs of thyrotropin-releasing hormone (TRH), obtained by the replacement of the L-pyroglutamic acid residue with 4-nitro- or 4-N butyloxycarbonylamino-1-methyl-2-pyrolylcarboxylic acid (analogs 1, 3), and three related derivatives, in which also the L-histidine residue was replaced with L norvaline (analogs 2, 4) or L-norleucine (analog 5), were synthesized and tested for endocrine and central nervous system (CNS) activity. The replacement of the L pyroglutamic acid residue with 4-nitro-1-methyl-2-pyrolylcarboxylic acid (analogs 1 and 2) resulted in the separation of the endocrine from the direct CNS activity. The effect of these analogs on the sleeping time, rectal temperature and breathing frequency, was either the same or greater than that of TRH. However, neither the correlation between the binding of analogs to TRH receptors in the brain nor their activity on the CNS parameters measured was found. Analogs 3, 4 and 5, containing 4-N-butyloxycarbonylamino-1-methyl-2-pyrolylcarboxylic acid in place of L-pyroglutamic acid, were inactive. PMID- 8401764 TI - Synthesis and cardiotonic evaluation of R-sulfonyl derivatives of azaheterocycles. AB - A series of alpha-sulfonyl-2-(3-chloro)hetarylacetonitriles was prepared by the interaction of dichlorodiazines and R-sulfonylacetonitriles. Reaction of nucleophilic substitution of chlorine by different N-H-azoles has been studied. The compounds obtained showed 3 types of activity on cardiac papillary muscles: 1. a biphasic activity with increase in cardiac contractility at low concentration and inhibition in high concentration (1, 2, 4, 12-15), 2. a monophasic inhibitory activity (3, 5, 8-10), 3. non-active compounds (6, 7, 11). The preliminary SAR studies indicate the importance of chlorine atom at position 3 in diazine nucleus and of alkyl substituents in benzimidazole moiety for positive inotropic effects. PMID- 8401765 TI - Disturbances in neurotransmission processes in aging and age-related diseases. AB - This paper reviews the changes in dopaminergic, cholinergic and glutamatergic neurotransmission, which occur in the aging of the central nervous system (CNS) and in age-related diseases: Parkinson's disease (PD) and Alzheimer's disease (AD). Dopaminergic neurotransmission is impaired with age due to degeneration of the substantia nigra pars compacta neurons and reduction of the density of postsynaptic D1 and D2 dopamine receptors in the striatum. PD is believed to be caused by a severe loss of dopaminergic neurons, which leads to nearly complete depletion of dopamine in the striatum, particularly in the putamen. The supersensitivity of postsynaptic dopamine receptors, reported by some authors, may result from compensatory mechanisms to degeneration of dopaminergic neurons. The role of aging in PD is also discussed in the paper. An interest in the role of the cholinergic and glutamatergic systems in aging results from the concept that the development of AD is due to the pathology of these systems. The data on cholinergic neurotransmission are controversial and imply that aging affects rather slightly both neurons and cholinergic receptors. In AD, however, severe degeneration of cholinergic neurons of the basal nucleus of Meynert, leading to the impairment of cholinergic neurotransmission in the hippocampus and the cerebral cortex, has been observed. In AD degeneration of glutamatergic neurons and subsensitivity of some excitatory amino acids receptors in the hippocampus and the cerebral cortex may lead to dementia. However, an increase in the glutamate release from presynaptic glutamatergic terminals may be responsible for neuronal degeneration in AD. The role of the beta-amyloid protein in a neurodegenerative activity of glutamic acid is discussed. PMID- 8401766 TI - The effect of repeated treatment with oxaprotiline enantiomers on central 5-HT receptor subpopulations. AB - The effect of repeated administration of (+)-OXA (a noradrenaline (NA) uptake inhibitor) and (-)-OXA (devoid of an effect on the NA uptake, but a clinically active antidepressant drug) on central 5-HT receptor subpopulations was studied. (-)-OXA given repeatedly, but not acutely, attenuated the 8-OH-DPAT-induced hypothermia in mice. (+)-OXA administered acutely, as well as repeatedly, was inactive in that test. The 8-OH-DPAT-induced syndrome in rats was attenuated by both OXA isomers administered either acutely or repeatedly. The hypothermia induced by m-CPP in mice was attenuated by single-dose administration of (+)-OXA and (-)-OXA; when given repeatedly, (+)-OXA increased the action of m-CPP. (-) OXA administered repeatedly was inactive in that test. Either single or repeated administration of (+)-OXA had practically no effect on the depression of exploratory activity induced by m-CPP. (-)-OXA administered acutely or repeatedly attenuated the effect of m-CPP in the same manner. Acute, but not chronic, administration of (-)-OXA reduced the number of head-twitch episodes induced by 5 HTP in mice. Repeated, but not acute, treatment with (+)-OXA attenuated the effect of 5-HTP. The obtained results indicate that (+)-OXA administered repeatedly increases the reactivity of 5-HT1B receptors, decreases the reactivity of 5-HT2 receptors, and has no effect on the reactivity of 5-HT1A- (pre- and postsynaptic) and 5-HT1C-receptors. (-)-OXA given repeatedly decreases the reactivity of presynaptic 5-HT1A receptors and has no influence on the reactivity of postsynaptic 5-HT1A-, 5-HT1B-, 5-HT1C- and 5-HT2-receptors. PMID- 8401767 TI - Discriminative stimulus properties of (+)-oxaprotiline in rats. AB - Using a drug discrimination procedure on a fixed ratio (FR) 10 schedule of reinforcement, rats were trained to distinguish between the stimulus properties of the selective inhibitor of noradrenaline uptake (+)-oxaprotiline ((+)-OXA) and saline. Fifty per cent of the rats that learned to reliably discriminate (+)-OXA from saline, reached high discrimination accuracy. In substitution testing, maprotiline (MAP) and desipramine (DMI) produced dose-related responding on the (+)-OXA lever. (-)-OXA failed to show substitution. The results of our findings demonstrate that (+)-OXA may be used as a discriminative cue in rats. The NA uptake inhibition seems to be involved in the effect of this drug. PMID- 8401768 TI - The effects of single and repeated administration of MAO inhibitors on acoustic startle response in rats. AB - In this paper the effects of single and repeated administration of brofaromine (BRO) and moclobemide (MOC)--monoamine oxidase A (MAO-A) inhibitors as well as deprenyl (DEP)--monoamine oxidase B (MAO-B) inhibitor on behavior of rats in acoustic startle response test were studied. For comparison the tricyclic antidepressant amitriptyline (AMI) was also studied. BRO given in a single dose of 10 mg/kg diminished the amplitude of acoustic startle response, while in a dose of 30 mg/kg had no effect. MOC diminished the amplitude when given in a single dose of 30 mg/kg, and had no effect in a dose of 10 mg/kg. Both MAO type A inhibitors diminished the startle reaction amplitude when tested 2 h after the last dose of a repeated treatment (twice daily for 14 days, 10 or 30 mg/kg, po). 5 days after the last dose the treatments did not affect the response. DEP given both in a single dose of 0.25 mg/kg and repeatedly for 15 and 21 days, increased the amplitude of startle reaction immediately after completion of the treatment. 9 days after drug withdrawal this effect was not observed. DEP in a dose of 1.0 mg/kg had no effect on the amplitude of startle reaction in any experiments described above. The above results demonstrate that MAO-A inhibitors--brofaromine and moclobemide and a MAO-B inhibitor--deprenyl show opposit activity in acoustic startle response test. BRO and MOC exert the inhibitory activity, whereas DEP has an excitatory effect on the startle reaction. The reference drug, AMI had no effect when given in a single doses (2 and 10 mg/kg, po). After repeated treatment (twice daily for 14 days, 2 or 10 mg/kg, po), AMI diminished the amplitude of the startle reaction only in higher dose when tested 2 h after the last dose and had no effect in the experiment carried out 5 days after the last dose. PMID- 8401769 TI - Phorbol ester and central chemosympathectomy augment beta-adrenoceptor response by different mechanisms. AB - The aim of this study was to compare the mechanisms of increased responsiveness of the beta-adrenoceptor dependent cyclic AMP generating system induced by chronic decrease of noradrenaline availability (beta-upregulation) with that resulting from simultaneous stimulation of alpha-adrenoceptors (alpha potentiation) and to assess the role of protein kinase C in these phenomena. The beta-upregulation was produced by central chemosympathectomy with 6 hydroxydopamine. The role of alpha 1- and alpha 2-adrenoceptors was assessed by comparison of the effects of specific beta-adrenoceptor agonist isoproterenol with those of a mixed alpha-beta-adrenoceptor agonist noradrenaline, and clonidine was used to selectively stimulate alpha 2-adrenoceptors. The role of protein kinase C was assessed by measuring cyclic AMP responses in the presence and absence of 12-O-tetradecanoyl-phorbol 13-acetate. The results indicate that the mechanism of increased responsiveness induced by central chemosympathectomy is different from the alpha-potentiation, that only alpha 1-adrenoceptors are involved positively in alpha-potentiation, while the alpha 2-adrenoceptors play an inhibitory role, and that increased responsiveness following central chemosympathectomy may be inhibited by protein kinase C activation. PMID- 8401770 TI - The action of antidepressant drugs administered during calcium channel blockade. AB - This is an overview of the symposium concerned with the action of electroconvulsive treatment and some antidepressant drugs (mainly imipramine) applied during blockade of voltage-dependent calcium channels with nifedipine. The results in general suggest that a combination of calcium channel blockers with antidepressant drugs or ECT may be of clinical value in treatment of depression. PMID- 8401771 TI - Electroconvulsive shock-induced impairment of spatial learning is aggravated by nifedipine. AB - Chronic electroconvulsive treatment applied immediately after a training session or with a 15 min delay impairs spatial learning and memory in the Morris water maze paradigm, and this impairment is not counteracted, but rather aggravated by co-administration of a calcium channel blocker, nifedipine. PMID- 8401772 TI - The effect of chronic nifedipine and ECS in the forced swimming test in rats. PMID- 8401773 TI - The effect of calcium channel blockade during electroconvulsive treatment on cerebral cortical adrenoceptor subpopulations in the rat. AB - Chronic administration of nifedipine (5 mg/kg/day for 10 days) induced some biochemical effects consistent with those of antidepressants: a significant depression in cortical alpha 2-adrenoceptor density and reduction of beta adrenoceptor affinity; nifedipine co-administration with electroconvulsive treatment potentiated the beta-downregulatory effect of the latter. PMID- 8401774 TI - The effect of calcium channel blockade on the action of chronic ECT and imipramine on responses of alpha 1- and beta-adrenoceptors in the rat cerebral cortex. AB - Chronic co-administration of nifedipine and ECT or imipramine results in an increase in responsiveness of cerebral cortical alpha 1-adrenoceptors as measured by accumulation of inositol phosphate in cortical slices after noradrenaline stimulation; the responsiveness of beta-adrenoceptor, measured by accumulation of cyclic AMP, was depressed similarly by antidepressant treatment with and without nifedipine. PMID- 8401775 TI - The effect of chronic treatment with imipramine, ECS and nifedipine on the beta adrenergic function in the rat hippocampal slices. PMID- 8401776 TI - Chronic effects of imipramine and nifedipine on the beta-adrenergic function in neuronal and glial primary cultures. PMID- 8401777 TI - The effect of chronic treatment with imipramine on the G proteins mRNA level in the rat hippocampus--an interaction with a calcium channel antagonist. AB - The effect of long-term administration of imipramine (10 mg/kg po, twice daily, 30 days) with or without nifedipine (5 mg/kg ip, twice daily, 28 days) on the G protein alpha subunit, Gs alpha, Go alpha and Gi alpha mRNA levels was investigated in the rat hippocampus. An in situ hybridization histochemistry showed that imipramine decreased the Go alpha mRNA level in CA1 (by ca. 40%) and CA3 (by ca. 37%) hippocampal fields and, to a lesser extent, in the dentate gyrus (by ca. 25%), but had no effect on the Gs alpha and Gi alpha mRNA levels in those structures. Nifedipine decreased (by ca. 30%) the Gs alpha level in the studied fields of hippocampal formation, having no influence on the level of mRNA which codes other subunits of G protein. Coadministration of nifedipine and imipramine reversed the imipramine effect on Go alpha, but had no effect on the nifedipine induced decrease at the Gs alpha mRNA level. These results suggest that inhibition of L calcium channels modifies the effect of imipramine at the level of intracellular signal transduction. PMID- 8401778 TI - Dementia of frontal type. PMID- 8401779 TI - Frontal lobe degeneration of non-Alzheimer type revisited. AB - The neuropathology of frontal lobe degeneration of non-Alzheimer type was reevaluated on the basis of a new material of 13 cases against the background of experiences from earlier published 16 cases. The salient neuropathological feature was an unspecific neuronal degeneration of superficial cortical layers of the frontal and to some extent the temporal lobes without markers for Alzheimer's, Pick's or Lewy body diseases and there were no indications so far of a prion etiology. The consistency of histopathological features are taken to indicate a disease entity, also identified by other authors. It is tentatively grouped together with progressive and aphasic dementia, similar cases in the literature and possibly also Pick's disease and ALS with dementia. PMID- 8401780 TI - Overview of dementia lacking distinctive histology: pathological designation of a progressive dementia. AB - The group of progressive dementing illnesses that lack distinctive histologic features includes at least four variants: a cortical type, a thalamostriate type, a motor neuronopathy type and a leukogliotic type. While the clinical presentation of some cases is that of anterograde amnesia, progressive aphasia and dysexecutive syndrome are the most common initial symptom complexes. A large number of reported cases are familial, although no abnormal gene has been identified. Pathologically, these illnesses are defined by cortical, hippocampal, striatal, medial thalamic, nigral and motor nuclei cell dropout and astrogliosis. In some cases, white matter gliosis is striking. Identification of specific histological or molecular markers of at least some of these conditions will greatly advance our understanding of these specific conditions as well as dementing illnesses in general. PMID- 8401781 TI - Presenile dementia with motor neuron disease. AB - Seventy-one Japanese cases of presenile dementia with motor neuron disease were reviewed. The clinico-pathological features were: (1) progressive dementia with insidious onset, mostly in the presenile period: (2) neurogenic muscular wasting in the course of illness (ALS- or SPMA-like symptoms); (3) duration from the onset of the illness to death: 2-5 years (average 30.6 months); (4) extrapyramidal symptoms and definite sensory deficits are less commonly present; (5) no characteristic abnormalities in the CSF or EEG; (6) no known consanguinity or familial occurrence; (7) non-specific mild to moderate degenerative changes in the fronto-temporal cerebral cortex, hypoglossal nuclei and spinal cord, and frequently in the substantia nigra. The author was interested in discovering whether the frequency and topology of lesions in the brain of patients with presenile dementia and motor neuron disease differed characteristically from the distribution found in cases of Alzheimer's disease, Pick's disease, Creutzfeldt Jakob disease or progressive subcortical gliosis. Presenile dementia with motor neuron disease might be a new disease entity. PMID- 8401782 TI - Clinical picture of frontal lobe degeneration of non-Alzheimer type. AB - Frontal lobe degeneration of non-Alzheimer type (FLD) is the second most common primary degenerative dementia in southern Sweden. Clinical findings in 30 FLD cases with postmortem-verified diagnoses are described. FLD starts in the presenium with a mean disease duration of 7.5 years (range 3-17 years). Clinical onset is insidious and slow and the early stage is dominated by personality changes with lack of insight and judgement and signs of disinhibition. A typical feature is progressive loss of expressive speech with stereotyped phrases, late mutism and amimia. Restlessness, changes of oral/dietary behavior and utilization behavior are prevalent as also psychotic features. Temporal and spatial orientation are usually preserved for a long time in contrast to Alzheimer's disease. Dementia in FLD is similar to that of Pick's disease and ALS with dementia. Early recognition of FLD seems possible based on standardized clinical evaluation supported by neuropsychological tests, measurement of regional cerebral blood flow and other types of brain imaging. The etiology of FLD is unknown but a positive heredity was reported in 60%. PMID- 8401783 TI - Progressive frontal dysfunction. AB - A number of causes of progressive mental impairment show a preponderance of frontal lobe symptoms. It is proposed that these disorders can be subdivided into primary and secondary frontal dementias, based on the symptomatology and the site of major pathological disruption. An anatomical explanation, based on variations in cortical and subcortical influences, aids in understanding the clinical differences. PMID- 8401784 TI - The clinical pathological correlates of lobar atrophy. AB - Three clinical syndromes associated with fronto-temporal cerebral atrophy, studied in one centre are discussed: dementia of frontal type (DFT), DFT and motor neurone disease (MND) and progressive aphasia (PA). The pathological findings in DFT (13 brains), DFT and MND (5 brains) and PA (5 brains) permit a number of clinical pathological groupings. The nosological status of fronto temporal atrophy is discussed with reference to the literature and it is suggested that a common underlying pathology, including Picks disease as strictly defined by the presence of inclusion bodies, underlies the clinical syndromes, each being determined by the anatomical distribution of the pathology. PMID- 8401785 TI - Spectrum of frontal lobe dementia in a Swedish family. AB - The etiology of frontal lobe dementia of non-Alzheimer type (FLD) is still unknown. There is strong evidence of genetic factors with positive heredity. In this paper, a Swedish family, with several generations affected by FLD is described. In 3 patients typical FLD was confirmed postmortem. The clinical and neuropathological similarities between the patients are impressive. PMID- 8401786 TI - Exclusion mapping in familial non-specific dementia. AB - We present genetic linkage data in a large family in which non-specific dementia is inherited as an autosomal dominant trait. We have analyzed 45 highly polymorphic microsatellite sequences and excluded a quarter of the genome as the site of the pathogenic mutation in this family. PMID- 8401787 TI - Neuropeptides in cerebrospinal fluid of patients with Alzheimer's disease and dementia with frontotemporal lobe degeneration. AB - The two major primary degenerative dementias, dementia of Alzheimer type (DAT) and frontal lobe degeneration of non-Alzheimer type (FLD) have several clinical features in common but also many symptoms that differ. In a clinical material of 80 patients with either of the two forms of dementia (DAT = 39, FLD = 41) we have studied the levels of neuropeptides in the cerebrospinal fluid (CSF) in order to find biochemical markers for CNS affection. The dementia forms were evaluated by careful clinical analysis, psychometric testing and measurement of regional cerebral blood flow. Approximately one third of the subjects died during the completion of the study and neuropathology was performed, confirming the diagnoses. We observed reductions in the CSF levels of antidiuretic hormone and somatostatin in both DAT and FLD. A strong tendency to reduction was noted for neuropeptide Y (NPY). There was a correlation with the duration of disease demonstrating a significant reduction in NPY levels in subjects with DAT. Most notably there was a strong reduction in the levels of delta sleep inducing peptide (DSIP) in DAT cases only. The levels of DSIP in FLD were the same as in controls. The reverse was found for corticotropin releasing factor (CRF) which had a significant reduction in FLD patients but not in those with DAT. The present study indicates a difference in the CSF levels of neuropeptides, observations that these may serve as biochemical markers which differentiate DAT and FLD. PMID- 8401788 TI - Preliminary neurochemical findings in non-Alzheimer dementia due to lobar atrophy. AB - Non-Alzheimer's dementia due to lobar atrophy had choline acetyltransferase activities comparable with control rather than Alzheimer's disease values, based on 3 autopsy proven cases on Pick's disease and biopsies from 3 examples of dementia of frontal lobe type. Muscarinic cholinergic receptors were relatively spared only in Alzheimer's disease. Serotonin receptors were markedly reduced (based on Pick cases) whereas measures that reflected presynaptic serotonergic activity were either not affected or increased. Cerebrospinal fluid and brain tissue measurements suggested that inhibitory interneurones and dopamine release were relatively spared. There was no in vitro evidence of hypometabolism. PMID- 8401789 TI - Prion diseases in humans and their relevance to other neurodegenerative diseases. AB - Molecular genetics has led to considerable advances in our understanding of the transmissible spongiform encephalopathies. The identification of pathogenic mutations in the prion protein gene has enabled a molecular reclassification of the familial forms of these diseases, which may now be referred to as inherited prion diseases. Prion diseases of both humans and animals are associated with deposition of an abnormal isoform of a host-encoded protein, the prion protein (PrP). Human prion diseases have inherited, sporadic and acquired forms. A considerable body of evidence now supports the idea that the transmissible agent in these diseases may be an abnormal isoform of the prion protein. The identification of pathogenic mutations in the PrP gene has enabled the identification of cases of inherited prion disease that would not have been recognised using existing clinical and pathological diagnostic criteria. Since marked clinical and neuropathological overlap between the different neurodegenerative disorders is well recognised, PrP gene analysis is of increasing importance in differential diagnosis. Frontal lobe dementia of non Alzheimer type and Pick's disease share a number of important clinical and pathological features with prion diseases, and could be considered as candidate prion diseases. However, we have not been able to demonstrate either PrP mutations or the presence of the disease-associated isoform of prion protein in several well-characterised families with these disorders. PMID- 8401790 TI - Regional cerebral blood flow in frontal lobe dementia of non-Alzheimer type. AB - Twenty-five out of 26 cases of autopsy-verified frontal lobe degeneration of non Alzheimer type (FLD) were found to have focal frontal or frontotemporal blood flow reductions involving both hemispheres. The deviant case had an asymmetric frontal pathology only apparent on the right side. Focal reduction of blood flow in the frontal lobes is, however, a common and unspecific flow abnormality found in e.g. Pick's disease. Creutzfeldt-Jakob's disease, and in some cases of Alzheimer's disease. Low frontal flow has also been reported in schizophrenia and in toxic encephalopathy. Since a characteristic feature of FLD is a steady progress of the pathology, serial flow measurements extending over several years are especially informative. PMID- 8401791 TI - Functional activation of the frontal lobes. Regional cerebral blood flow findings in normals and in patients with frontal lobe dementia performing a word fluency test. AB - The present study examined the utility of the Word Fluency Test (WFT) as a frontal-lobe-activating test in brain imaging. Regional cerebral blood flow (rCBF) was measured during rest and during the WFT in 49 healthy volunteers and in 15 patients with frontal lobe dementia (FLD). The results showed a highly significant frontal lobe activation in 85% of the normal subjects. This finding was not related to age or to the level of performance on the WFT. A significant frontal activation was seen in 13 of the 15 FLD patients. The frontal flow increase did not reach normal levels, and was not related to age, illness duration or severity of clinical symptoms. The results suggest that the WFT is an ideal test to use in conjunction with functional imaging in normals as well as in patients with organic dementia. PMID- 8401792 TI - Functional imaging, the frontal lobes, and dementia. AB - A 58-year-old man developed progressive difficulty with comprehension and verbal output with dementia. Positron emission tomography with 18F 2-fluoro-2-deoxy-D glucose demonstrated asymmetrical frontal and anterior temporal lobe loss of glucose use. Scopolamine infusion (0.3 mg) did not influence memory. Postmortem studies revealed evidence of Pick's disease, with Pick bodies, loss of somatostatin, preservation of choline acetyltransferase and immunostaining with neurofilament antibodies. Pharmacological challenge and positron imaging offer valuable means for the noninvasive assessment of dementing illness. The contributions of functional imaging to our knowledge of frontal involvement in dementing illness are reviewed. PMID- 8401793 TI - Progressive right frontotemporal degeneration: clinical, neuropsychological and SPECT characteristics. AB - The behavioral, neuropsychological and single photon emission computerized tomography characteristics of 5 patients with progressive degeneration of the right hemisphere are described. In all, the brain regions with greatest involvement were right-frontal and temporal. Psychosis, compulsions and behavioral disinhibition were the dominant, and often first, symptoms. Affect was flattened and the patients seemed distant and remote. Neuropsychological testing did not reveal a consistent pattern that helped localize the abnormality to the right frontotemporal region. These patients contrast dramatically to those with left frontotemporal degeneration in whom behavior and psychiatric status is often normal. This study suggests that the right hemisphere may be primary for the control of social conduct. PMID- 8401794 TI - Neuropsychological findings in frontal lobe dementia. AB - Neuropsychological investigations were performed on 18 patients with a clinical diagnosis of frontal lobe dementia supported by regional cerebral blood flow measurements. Nature and degree of cognitive impairment were examined with a comprehensive test battery. The results of the neuropsychological assessment could be described as three levels of cognitive impairment. The increasing levels of cognitive impairment were accompanied by corresponding levels of reduced cerebral blood flow in frontotemporal areas. No apparent relationship emerged between impairment level and illness duration, indicating a considerable individual variation in the clinical course of frontal lobe dementia. PMID- 8401795 TI - Assessment of neuropsychological dysfunction in frontal lobe degeneration. AB - The quality of the neuropsychological investigation of behavioural changes in frontal dementias is dependent on the specificity of the behavioural analyses. A review of methodological issues and published findings identifies possible potential reasons for misinterpretation of research neuropsychological data in these studies. The areas reviewed include the operational definitions of terms such as 'frontal lobes' and 'frontal functions'; identification of experimental limitations in frontal lobe research; and highlights of neuropsychological investigations of frontal lobe functions. These deliberations suggest a practical approach to the behavioural assessment of frontal dementias. PMID- 8401796 TI - Progressive language dysfunction and lobar atrophy. AB - The patterns of language disorder associated with 'progressive aphasia' due to lobar atrophy were compared with the language dysfunction of patients with dementia of frontal lobe type (DFT). The progressive aphasias were characterised primarily by impairment at the structural levels of language: phonology, grammar and semantics, whereas DFT was associated primarily with spontaneity and loss of generative capability. However, there was overlap in language symptomatology, particularly with progression of disease. The findings lend support to the argument that progressive aphasia and DFT represent different clinical manifestations of a common pathology, and form part of the spectrum of lobar atrophies. PMID- 8401797 TI - Frontal lobe cognitive functions in aging: methodologic considerations. AB - The empirical literature has been contradictory regarding whether frontal functions decline with age, perhaps due to important differences across studies in medical and psychiatric exclusion criteria. We review literature and present preliminary data suggesting that frontal lobe functions are particularly affected by several medical and psychiatric disorders common in old age. Investigations of frontal lobe functions in aging require careful screening of subject samples; otherwise, any impact of age on cognition is contaminated by the effects of coexistent physical and/or psychiatric illness. PMID- 8401798 TI - Peripheral nerve injuries: pathophysiology and strategies for treatment. PMID- 8401799 TI - Functional comparison of pronation and supination strengths. AB - Studies have shown that functional activities involving pronation and supination, such as turning a doorknob or a screwdriver, involve wrist positions of extension and/or ulnar deviation. Previous studies of isometric forearm pronation and supination strength have utilized a neutral wrist position, resulting in a possible underestimation of true functional strength. Twenty healthy subjects between the ages of 24 and 45 years were assessed for isometric pronation and supination strength using cylinder, screwdriver, and doorknob handle adaptations. Maximal strength in both pronation and supination was observed with the use of the doorknob handle. Although pronation strength was exceeded by supination strength within all three handle adaptations, the difference was statistically significant for the screwdriver and doorknob handles. The results suggest that in clinical and research-oriented testing of maximal forearm rotation strength, further consideration must be given to the terminal grip device and positioning of the wrist. PMID- 8401800 TI - The relationship between two power-grip testing devices and their utility in physical capacity evaluations. AB - This study compared two isometric-grip testing devices and considered the efficacy of combining data from the two as a means of identifying both functional status and consistency of effort for physical capacity evaluations (PCEs). Two groups (26 healthy subjects and 40 patients who had upper-extremity impairments) were used to establish the reliability of applying force on a jamar dynamometer, and on an Isometric Strength Testing Unit (ISTU). Many of the patients had been referred because of inconsistencies between subjective complaint and medical presentation. The results indicate that power-grip force outputs are highly related across the two test devices, which validates their utility as part of a PCE for wrist-hand involvement. A simple index (2-ratio) of functional status and consistency was developed by combining test data from the two devices. The sensitivity of the 2-ratio, in combination with other indices of consistency, is discussed. PMID- 8401801 TI - The effect of water temperature on hand volume during volumetric measurement using the water displacement method. AB - This study evaluated the effect of water temperature on hand volume during volumetric measurement using the water displacement method. Four volumetric measurements were taken for 24 normal subjects using four different water temperatures--5 degrees C (41 degrees F), 20 degrees C (68 degrees F), 35 degrees C (95 degrees F), and 45 degrees C (113 degrees F). Between volumetric measurements the skin temperature of the hand was allowed to return to baseline levels using an Orion biofeedback unit. In comparing the two extreme temperatures, the average change in hand volume was +5.8 milliliters (ml), which represented a deviation of 1.4% of the mean (coefficient of variation). In using a t-test to determine significance, it was found that the change in volume between these two temperatures was significant at the 0.01 level. In comparing the two moderate temperatures, the average change in hand volume was +1.9 ml, representing a deviation of 0.5% of the mean, with no statistical significance. For research purposes, this study suggests the need to control for water temperature when using a volumeter to measure hand edema. For general clinical practice, however, the use of "cool" or "tepid" water does not appear to alter hand volume readings significantly to be of concern. PMID- 8401802 TI - Maximizing therapist effectiveness with geriatric hand patients. AB - The "graying of America" is resulting in higher proportions of older persons utilizing the health care system. Current trends indicate that the geriatric population represents a growing portion of the hand therapy patient population. Appreciating the unique needs of the geriatric hand patient and developing an appropriate treatment program require special interventions that may seem unappealing due to the likelihood of prolonged initial treatment time. However, such steps can strongly enhance therapist effectiveness, patient compliance, and ultimate outcome. The purposes of this paper are: (1) to address frustrations that may be encountered in the treatment of some elderly hand patients; (2) to introduce the reader to the phenomenon of age bias; (3) to present a well accepted and effective assessment tool of memory, orientation, and ability to follow commands as these competencies pertain to hand therapy; (4) to address concepts in the gerontology literature that can be applied clinically as treatment guidelines to boost treatment effectiveness; and, finally, (5) to suggest communication techniques to use with the elderly hand patient. Maximizing therapist effectiveness with geriatric hand patients will serve to reinforce the dignity and value of this unique population. PMID- 8401803 TI - Who should do research? PMID- 8401804 TI - The rheumatoid hand: postoperative splint options. PMID- 8401805 TI - Modified arm sling for soft-tissue injuries of the shoulder and glenohumeral subluxation. PMID- 8401806 TI - Techniques for improving distal interphalangeal motion. PMID- 8401807 TI - Defining performance of organizations. AB - Quality is and always has been in the beholder's eye. It is therefore necessary, as argued in this excerpt from The Measurement Mandate, the latest text on the Joint Commission's Agenda for Change indicator development and testing methodology, to concentrate on the best ways to measure, assess, and improve the discernible, quantifiable dimensions of an organization's performance. PMID- 8401808 TI - Empowering health care improvement: an operational model. AB - BACKGROUND: The shift from compliance to continuous improvement requires a bridge -empowerment--that enables the creation of an environment in which employees can contribute the full range of their expertise in service to the organization. Leaders are responsible for creating this empowering environment. MODEL: An operational model that describes the nine aspects of empowerment (for example, mission and values, access to information, avenues of influence) can serve as a template for leaders to use to promote an atmosphere of empowerment in their organizations. METHOD: The authors adapted this model as a survey instrument to uncover some hospital success stories that demonstrate several aspects of empowerment in action. The survey was administered at two hospitals. CASE STUDY 1: At Beth Israel Hospital (Boston, Mass), Nutrition Services rated the "mission and vision" aspect of empowerment in their department at an average of 3.5 out of a possible 4.0. The department attributes this high rating to their focus on specific goals that reinforce the larger mission and vision of the hospital as a whole. These goals form the basis for annual employee evaluations. For example, a 1992 patient-focused goal was to provide nutrition counseling and education. The department approached this goal by developing a daily format ("Eat Smart") for the daily hospital cafeteria menu that provided nutritional information. The rationale was to educate employees, who could then bring back information to their patients. Alignment of the department's efforts with the larger mission of the hospital enabled Nutrition Services to extend its influence beyond departmental and professional boundaries. Implementation of other plans at the hospital demonstrate a sustained process of vision development on the part of the executive leadership. CASE STUDY 2: At Robert Wood Johnson University Hospital (New Brunswick, NJ), Support Services gave the "environment of trust and respect" aspect an average rating of 3.7. The key to this success story lies in the upgraded role of the housekeepers as "hosts." The "Pro-Act" restructuring model for patient care delivery creates and upgrades positions to relieve nurses of their task loads. The three nursing services that participated in the survey gave their highest rating to the "access to the education and training" aspect of empowerment. The high quality of professional excellence is supported by tuition reimbursement, incentives for credentials, the extern program and scholarships provided to attract outstanding nursing students to their staff and encouragement of nursing staff at all levels for publication, presentations, and poster sessions. CONCLUSION: These case studies demonstrate how leaders created an environment in which collective learning has become a way of doing business. Given the multifaceted nature of the empowerment model, some may wonder where to begin to focus leadership efforts. It is important to emphasize that the constellation of empowerment aspects constitutes an integral system; the full potential for empowerment arises from synergy among all nine aspects. PMID- 8401809 TI - Critical path method: an important tool for coordinating clinical care. AB - BACKGROUND: In May 1991 Mount Clemens General Hospital (MCGH) began investigating the critical path method (CPM) as a tool for extending total quality management in clinical areas. In its search for guidelines on how to develop a critical path program, it found that other hospitals used a variety of approaches. These included employing case managers or outside consultants to develop programs or implementing prepurchased paths. Because these approaches often are difficult to customize for a specific institution and because MCGH wanted to use an internal team, none of these options seemed appropriate. With no definitive guidelines to follow, MCGH developed and implemented its own CPM. METHODS: The developmental process was composed of activities in nine primary categories: literature search, steering group, targeting strategy, paperwork design, gaining consensus, pilot program, preliminary findings, refine program, and full implementation. RESULTS: A pilot was performed to assess if the CPM would be beneficial. Six months into the pilot a preliminary review of coronary artery bypass graft paths was conducted. There were 44 patients in the study group (35 men, 9 women). Twenty four patients were cared for before the critical path form was available. Preliminary findings indicated a lower rate of complications in patients cared for with the critical path form. The data revealed a 5% complication rate with the critical path compared to a 16.6% rate for those whose care was not guided by the form. In addition, patients with the path on their clipboards had an overall shorter length of stay than patients without the path. It is important to remember that these early data are based on the six-month pilot; they are not considered a conclusive research finding. NEXT STEPS: The next step in the CPM process is to examine other diagnoses that might benefit from a critical path approach. A steering committee composed of representatives from hospital administration, nursing, medical staff, quality assurance and risk management, and total quality management will act as the approval body for investigating and sanctioning other paths for development. CONCLUSION: The primary lesson learned at MCGH is that the CPM is most effective in an environment of communication and commitment. This approach allows clinic and nonclinic staff to talk about how their work influences each other's. CPM provides all caregivers with a common language and encourages everyone to look at the whole patient and the entire care process. The key message of success is: Get a group of people together who are motivated and empowered to move this exciting tool of the future through the necessary steps. PMID- 8401810 TI - Development of outcome-based practice guidelines: a method for structuring problems and synthesizing evidence. AB - BACKGROUND: The growth in guidance development projects has focused attention on the methods used in developing the guideline. For a guideline to be sound it should be linked on the basis of scientific evidence to the very health outcome that the guideline is designed to promote. METHODS: Structuring a health intervention as an influence diagram, a decision model (1) allows for the identification of the relevant benefits, harms, and costs that may result from an intervention; (2) provides an explicit link between the intervention and these outcomes, a crucial prerequisite for the development of an outcome-based guideline; and (3) identifies the evidence that must be synthesized to predict the effect of the intervention on the health outcomes. EXAMPLE: In the development of a guideline related to prevention of opportunistic infections in HIV-infected persons, we would define the interventions (for example, use of medication for PCP pneumonia), the intended health outcome (a potential reduction in the number of opportunistic infections), and the evidence that demonstrates that the intervention produces the desired outcome. If PCP prophylaxis is delayed, the HIV-infected person is exposed to a undue risk of PCP, with its attendant morbidity and mortality. If it is initiated too early, the person incurs excess monetary costs and may experience additional side-effect-associated morbidity. EXAMPLE: The intervention in question is screening for HIV infection, and the outcomes of interest are the medical benefits and harms associated with screening and the financial costs (and savings) that a screening program would incur. Screening for HIV infection differs from many clinical questions because it has potential benefit both to the persons screened and to public health if the screened person reduces risk behaviors that might transmit HIV infection. CONCLUSIONS: Structuring a problem with an influence diagram: delineates an explicit link between interventions and outcomes; focuses the questions to be addressed (a series of more sharply defined questions, each of which we may be able to answer based on direct evidence, replaces a much broader question [should we screen for HIV?], which cannot be answered directly); and highlights the importance of a clear, unambiguous statement whose benefit and costs are under consideration. PMID- 8401811 TI - Gathering systematic information on health plans: an interview with Sheila Leatherman. Interview by Elizabeth A McGlynn. PMID- 8401812 TI - Fetal rat pancreas transplantation in BB rats: immunohistochemical and functional evaluation. AB - Spontaneously diabetic BB/Wor rats received either a syngeneic fetal pancreas transplant or adult islets. In the former, 4-8 fetal pancreases were transplanted, and in the latter, 3-5000 islets. Transplantation was performed by transferring a blood clot containing the pancreases or islets to the renal subcapsular space. Insulin therapy was undertaken postoperatively, except in one experiment with adult islets. Of the fetal pancreas transplanted BB rats, 52% became normoglycaemic, and 21% remained so throughout an observation period of 10 months. Nephrectomy caused a prompt return of diabetes. The histological appearance of the grafts transplanted to the diabetic animals closely resembled that of grafts transplanted to normal rats in a parallel series. For comparison a group of BB rats received a syngeneic transplant of isolated adult islets from WF rats or BBW rats. Following adult islet transplantation, 5 out of 6 animals became hyperglycaemic after a median of 20.5 days when no insulin was given post transplantation. Four out of 5 animals became hyperglycaemic after a median of 23 days when supportive insulin therapy was administered after the transplantation. The results indicate that recurrent diabetes is not inevitable following syngeneic fetal pancreas transplantation to spontaneously diabetic BB rats. Recurrent diabetes was only occasionally associated with mononuclear cell infiltration. Transplanted tissue was well-preserved and vascularized; mega islets were a constant finding. PMID- 8401813 TI - Smooth muscle differentiation in human myometrium and uterine leiomyoma. AB - Smooth muscle differentiation has been analysed in human myometrium and leiomyoma by Western blotting with antibodies to smooth muscle specific proteins. No differences in the expression of h-caldesmon, metavinculin, desmin, alpha-smooth muscle actin and calponin were observed. The technique of two-dimensional gel electrophoresis was used, therefore, to further analyse differences between normal smooth muscle cells and their neoplastic counterparts. By comparing the protein patterns of normal myometrium and leiomyoma, it was possible to identify a protein with a molecular weight of approximately 27 kD that is selectively expressed in normal uterine smooth muscle cells. This protein proved to be a low molecular weight variant of calponin, a smooth muscle specific protein of as yet unknown function. Its immediate downregulation in tissue culture of normal myometrium points to a possible role in the process of dedifferentiation. PMID- 8401814 TI - The effects of obstruction and secretory stimulation on microlithiasis in salivary glands of cat: light and electron microscopy. AB - Obstruction and increased secretory activity are considered to be important aetiological factors of salivary microlithiasis, which may itself be an aetiological factor of sialadenitis. However, there is a lack of substantial evidence for the importance of obstruction, and investigations on increased secretory activity used pathological doses of pharmacological agents. Therefore further investigation of these factors is essential. Feline parotid, submandibular and sublingual salivary glands, in which microliths occur normally as in man, were examined after ductal ligation to produce obstruction, electrical stimulation of the glandular nerves to produce increased secretory activity, or both. Microliths were detected in: 0 out of 38 untreated, 1 out of 55 ligated, 2 out of 17 stimulated and 2 out of 17 stimulated ligated submandibular glands; 6 out of 29 untreated, 7 out of 46 ligated, 3 out of 12 stimulated and 4 out of 14 stimulated ligated sublingual glands; and no parotid glands. The chi 2 test confirmed that the experimental procedures did not produce an increased occurrence of microliths. Microliths were detected in parenchymal cells, intercellularly in atrophic parenchyma, intraluminally, interstitially and in macrophages. The present investigation indicates that obstruction and increased secretory activity are not important aetiological factors of salivary microlithiasis. PMID- 8401815 TI - Gastrointestinal AAPOAII and systemic AA-amyloidosis in aged C57BL/Ka mice. Amyloid-type dependent effect of long-term immunosuppressive treatment. AB - The light microscopic and immunohistochemical features of a novel localized senile amyloidosis in the gastrointestinal tract of C57BL/Ka mice are described. Senile gastrointestinal amyloidosis was predominantly found in the lamina propria of the ileum, cecum and stomach and infrequently in other segments of the gastrointestinal tract. The Congo red affinity of the senile amyloid was sensitive to potassium permanganate pretreatment. The amyloid did not react with anti-AA and anti-immunoglobulin antisera, but stained positively for apoAII, a major apolipoprotein of high density lipoproteins. A similar type of amyloid, termed AApoAII, has recently been described in a systemic form of senile amyloidosis in mice. In the present study, we investigated the effect of long term immunosuppressive treatment on the incidence of systemic AA-amyloidosis and gastrointestinal AApoAII-amyloidosis in aged C57BL/Ka mice. Gastrointestinal amyloidosis occurred in 60% of the control mice, but significantly less in mice of the immunosuppressed groups. In contrast, systemic AA-immunoreactive amyloidosis was only found in mice that were given immunosuppressive treatment. There was no codeposition of AA and AApoAII-amyloid. These findings indicate that immunosuppressive drugs have a profound effect on the incidence as well as the type of amyloidosis in C57BL/Ka mice. PMID- 8401816 TI - Inhibition of the effects of rheumatoid synovial fluid cells on chondrogenesis and cartilage breakdown in vitro: possible therapeutical conclusions. A morphological--biochemical study. AB - Short-term co-cultivation of blastemal cells from 12-day-old mouse limb buds and human rheumatoid synovial fluid cells in high density cultures (Trowell culture system) resulted, depending on when co-cultivation started, either in (1) an inhibition of chondrogenesis (co-cultivation right from the start) or in (2) an extensive breakdown of cartilaginous matrix (co-cultivation after formation of embryonic cartilage). These synovial effects were markedly impeded if Avarol (a dioxygenase inhibitor) was applied singly or in combination with PAI-2 (a u-PA inhibitor). PAI-2 alone, however, had no effect on the synovial-induced inhibition of chondrogenesis, but produced a pronounced inhibitory effect on matrix breakdown. The effects of both inhibitors were studied electron microscopically and biochemically (determination of sulfated-glycosaminoglycans in the high density cultures by Alcian Blue binding assay). The results of this study are consistent with the presumption that rheumatoid synovial cells are capable of inhibiting chondrogenesis and enhancing the breakdown of the cartilaginous matrix. Amongst others, the possible mediators involved are prostaglandins and plasminogen activators. The response to the inhibitors Avarol and PAI-2 is compatible with their mode of action. The chondroprotective action of these substances may be useful in developing potential antirheumatic drugs. PMID- 8401817 TI - Ultrastructural study of hepatic granulomas induced by Cryptococcus neoformans by quick-freezing and deep-etching method. AB - The ultrastructure of hepatic granulomas induced by Cryptococcus (C.) neoformans was studied by a quick-freezing and deep-etching (QF-DE) method. Viable yeast cells were inoculated intravenously into rats and the livers were prepared for QF DE replicas. Two cytoskeletal components were identified in the cytoplasm of macrophages composing the cryptococcal granulomas. These were: intermediate filaments, mainly located in the perinuclear region, and actin filaments, which were extensively decorated with myosin subfragment 1 (S1) and formed networks in the peripheral portion of the cytoplasm. In addition, two types of macrophage pseudopodia were observed in the granulomas. These were cobble stone-like pseudopodia at the yeast-macrophages contract areas, and thin, long and occasionally interdigitating pseudopodia in which actin filaments were consistently observed. Dense networks of actin filaments were also seen in pseudopodia protruding into the tight structure of the capsule of C. neoformans. These results suggest a role for actin filaments as one of the main factors in the force generating system of the phagocytic process. PMID- 8401818 TI - Peroxisomes in liver, kidney and duodenum of nude mice bearing xenografts of human pancreatic adenocarcinomas. AB - In the liver, kidney and duodenum of nude mice with xenografts of two human pancreatic adenocarcinomas differing in growth rate, catalase activity was assayed and peroxisomes were studied using catalase cytochemistry and light and electron microscopy. Hepatic and duodenal catalase activity were significantly decreased in tumour-bearing mice. Renal catalase activity was unchanged. At light microscopic level, a decrease in peroxisomal staining was evident in all duodenums and most livers of tumour-bearing mice. Only minor changes were observed in the kidneys. Ultrastructural morphometry of the hepatocellular peroxisomes revealed a decrease in size, volume density and surface density only in mice with fast-growing xenografts. These observations indicate that the two pancreatic adenocarcinomas exerted a different effect on the hepatic peroxisomes, and that catalase activity and peroxisomes in liver and duodenum are more affected than in kidney. PMID- 8401819 TI - The Food and Drug Administration and its influence on home bleaching. AB - The era of bleaching vital teeth has captured the attention of the dental profession, the public, the media, and the government. This method, using a custom-fitted mouthguard and a carbamide peroxide solution, is known as home bleaching, matrix bleaching, nightguard vital bleaching, passive bleaching, and dentist-prescribed-home-applied bleaching. Recent action of the US Food and Drug Administration and continued research and clinical experience in the area have provided favorable and unfavorable information about the variations of the technique. This article discusses these variations, with the general conclusion that the technique of vital tooth bleaching, when administered by a dentist using a custom-fitted mouthguard, is as safe as many other routinely performed dental procedures. PMID- 8401820 TI - Resin bonding to various substrates. AB - The use of bonding adhesive techniques in dentistry in general and in esthetic dentistry in particular has expanded significantly. As techniques, materials, and results improve, adhesive procedures are substantially modifying many traditional approaches to treating dental problems. More esthetic and less invasive techniques are becoming routine as adhesive materials and techniques become tested and understood. This paper reviews several important areas that affect our understanding and application of adhesive dentistry. With the many material categories, materials in categories, and varieties of bonding surfaces, the subject is extremely complicated. An attempt has been made to divide the past year's literature into sections and to review each section as a separate component of this complex subject. PMID- 8401821 TI - Current state-of-the-art porcelain veneers. AB - The literature is increasingly describing the use of resin-retained porcelain veneers. The dental profession has eagerly adopted this technique and much of this interest has been fueled by the influx of patients seeking to improve the appearance of their smiles. Properly performed, porcelain veneers can provide outstanding esthetics, strength, and longevity. Important restorative applications are beginning to emerge that broaden the scope of ultra-thin bonded ceramic techniques. As with any new procedure, clinical and scientific investigation is required to determine the parameters for efficacy. This review will highlight some of the recently published papers that address important issues associated with the use of porcelain veneers. PMID- 8401822 TI - Advances and current research in ceramic restorative materials. AB - Ceramics are currently the most widely used material for veneering crowns and bridges. Newer applications, such as bonded inlays, onlays, and veneers, are steadily gaining in popularity. A new generation of metal-free, all-ceramic systems for crowns became available during the 1980s. More recently, other all ceramic systems with improved mechanical properties affording a variety of uses have been introduced. Also, a computer-assisted design-computer-assisted manufacture method of fabricating ceramic inlays has become a routine treatment entity with favorable 5-year clinical data. Research is leading to a better understanding of conventional ceramic systems as well as providing information on laboratory manipulation and clinical methodology for optimal results. PMID- 8401823 TI - Inlays and onlays. AB - Dentists must meet the increased demands of patients for esthetic dentistry. Because the substitution of amalgam with direct composite restorations is somewhat problematic and restricted to special indications only, the most suitable method is the esthetic inlay. All available systems are reviewed and described. These restorations must be inserted with adhesive techniques. The performance and the problems associated with these techniques are described in detail. Improvements in luting techniques, eg, the use of ultrasonic energy, may further simplify the application technique for esthetic inlays. For clinical success, the margin quality of adhesively luted inlays is essential. In vitro and in vivo studies with esthetic inlays report good clinical behavior of such restorations. However, until the results of long-term clinical studies are available, these systems should not be used on a routine basis as a substitute for amalgam. PMID- 8401824 TI - High technology in esthetic dentistry. AB - Unimagined only a few years ago, high-technology tools for the dental practice are now widely available. Ranging from video cameras to computer-enhanced diagnostics that fabricate porcelain restorations on site, these new technologies have many applications in esthetic dentistry. Because they make it easier to communicate to patients what can realistically be achieved, high-technology tools tend to involve the patient more in treatment planning and thus may help prevent disappointing outcomes. PMID- 8401825 TI - From function to esthetics: anterior or occlusal compromises to esthetics. AB - Increased patient expectations together with improved diagnostic, material, and surgical advances have expanded the boundaries of esthetic dentistry. To optimize functional and esthetic success, existing techniques are being enhanced through careful patient selection and management of the occlusion. During the period from 1991 to 1992, several areas relating anterior or occlusal compromises to esthetics have been identified. These include: dental and skeletal malocclusion, periodontal esthetic defects, restorative materials and laboratory techniques, tooth arrangement and maxillomandibular relations, and implant dentistry. This paper reviews some of the literature on these areas during this period. PMID- 8401826 TI - Periodontal considerations for esthetics: edentulous ridge augmentation. AB - Edentulous ridge augmentation is a plastic surgical technique that is performed to improve patient esthetics when unsightly, deformed ridges exist. This article describes the etiology of ridge deformities and the many procedures that can be executed to achieve an esthetic, functional result. Historically, soft-tissue mucogingival techniques were described to augment collapsed ridges. Pedicle grafts, free soft-tissue grafts, and subepithelial connective tissue grafts are predictable forms of therapy. More recently, ridge augmentation techniques were developed that regenerate the lost periodontium. These include allografts, bioglasses, guided tissue regenerative procedures, and tissue expansion. PMID- 8401827 TI - Esthetic considerations in adult orthodontic treatment. AB - The use of orthodontics to correct dental malocclusions and enhance the cosmetic appearance in the adult patient is evaluated. A review of the current literature highlights the latest materials and techniques in adult orthodontics. A categorization of the diagnostic variables involved and the therapeutic options available is presented. Alignment, sagittal, vertical, transverse, and soft tissue discrepancies, as well as future considerations, are discussed. PMID- 8401828 TI - Dentures and partials or esthetic removable prosthetics. AB - In the past 50 years, advances in restorative dental procedures have made it possible for dentists to achieve better esthetic and functional results for their patients than ever before. Removable prosthodontics has been practiced in much the same way that is has been for many years. Since the introduction of root-form osseointegrated implants, there is a renewed interest in the edentulous and partially edentulous patient. To give the patient all possible treatment options, implants must be considered, but they must not replace sound prosthodontic principles. Presurgical prosthodontics in the form of provisional restorations and stents are necessary for correct implant placement. Overdentures, bar partial dentures, and soft liners are sound treatment alternatives. Review of the work of Dr. Earl Pound is the best source for the development of correct complete dentures. PMID- 8401829 TI - Cosmetic oral and maxillofacial surgery: a new era. AB - Oral and maxillofacial surgery is a specialty of dentistry that may be likened to the proverbial "sleeping giant." From a humble, nascent beginning as a specialty to advance the then modern concept of alveolar surgery to its present state, oral and maxillofacial surgery is today vibrating with excitement. To the credit of dentistry, the profession now has a specialty that not only has the capability to alter the bony facial framework, but also can alter the collateral collagenous and superficial cutaneous structures. Combined with dental alteration of the dentition and gingival apparatus, the profession is nearing its zenith relative to having the capability of changing an entire face. PMID- 8401831 TI - Bibliography of the current world literature. Cosmetic dentistry. PMID- 8401830 TI - Dental implants. AB - The use of dental implants in the oral rehabilitation of partially or fully edentulous patients has become a frequently used treatment modality in everyday practice. The main reason for this development is that critical principles for implant predictability have been established in the last twenty years. Today, researchers and clinicians of all dental subspecialties including cosmetic dentistry are working at refining techniques and extending the limits of the field. This paper is intended to point the reader to interesting publications from 1991 through 1992. PMID- 8401832 TI - Root resection and root amputation. AB - Attachment loss in furcal areas is a challenge for the clinician. Root resection and root amputation techniques have been used to overcome the instrumentation problems that these areas pose. Recent reports indicate that this approach is effective and reliable but extremely technique-sensitive and expensive for endodontic and prosthetic involvement. A more conservative approach, such as scaling and root planing, is suggested only for shallow class II furcation. Guided tissue regeneration in deep class II and class III furcation seems to be unpredictable and the efficacy of this therapy in such areas is still questionable. PMID- 8401833 TI - Genetic and inheritance considerations in periodontal disease. AB - Considerable evidence suggests that there is some genetic basis for the early onset forms of disease. The molecular abnormality and its genetic inheritance has been established in some cases of generalized prepubertal periodontitis. Family studies of juvenile periodontitis indicate that this disorder is transmitted by autosomal recessive genes, although additional data indicate genetic heterogeneity in this clinically defined disease. Recent evidence also suggests that susceptibility to periodontal disease may be related in part to genetically determined immune responsiveness to bacterial lipopolysaccharides. Although specific genetic risk factors have not been identified for the more common adult chronic periodontitis, recent studies indicate that there is significant genetic variance in the population for clinical and radiographic measures of disease. More precise definitions of disease phenotypes will facilitate future genetic epidemiologic studies of the periodontal diseases. PMID- 8401834 TI - Periodontal regeneration. AB - This paper reviews the current clinical and histologic literature in the field of periodontal regeneration. Various procedures and agents that have been used to attain regeneration and new attachment are reviewed, including open flap debridement, root conditioning, the use of bone grafts and bone substitutes, guided-tissue regeneration, coronally positioned flaps, and a combination of these. Future directions for clinical and research efforts are discussed, with emphasis on growth factors, their delivery, and enhancement of the healing response. Literature identifying factors that will aid in the regeneration of lost periodontium is reviewed and critiqued. PMID- 8401835 TI - Biologically active factors in the treatment of periodontal disease. AB - Cells in normal tissue progress through programs of growth, differentiation, and death. Many of the signals that start and stop this process are initiated by growth factors produced by other cells or by the resident cells. This network of intercellular signaling represents the basis for establishing and maintaining normal tissue architecture. Growth factors are released by some cells and conveyed to target cells, where they are bound to specific receptors, triggering a complex signal transduction cascade. Human periodontal ligament cells have been shown by in vitro assays to respond both mitogenically and chemotactically to a variety of growth factors including, but not limited to, platelet-derived growth factor, basic fibroblast growth factor, and transforming growth factor-beta. Evidence is presented for confirmation of an autocrine chemotactic and mitogenic human periodontal ligament cell-specific factor. This type of knowledge suggests that future periodontal regeneration procedures will rely on the exogenous addition of specific growth factors. PMID- 8401836 TI - Periodontal plastic surgery. AB - Mucogingival surgery has evolved in the past decade into periodontal plastic surgery. Periodontal plastic surgery deals with those regenerative and reconstructive procedures designed not only to restore form and function but also to enhance esthetics. This paper reviews those techniques that currently comprise periodontal plastic surgery as well as the current literature pertinent to this topic. The role of the dental laser as well as its limitations will also be discussed. PMID- 8401837 TI - Future directions of lasers in dental medicine. AB - Dental practitioners continue to serve as leaders in the development of laser applications for dental surgery. Laser wavelengths ranging from the ultra-violet excimer lasers to the far-infrared carbon-dioxide lasers are now used in dental research with greater frequency. By using new and emerging laser media, researchers can now demonstrate advances in the fields of laser diagnostics, photocoagulation, photoablation, and laser tissue-welding. Dental laser therapy is rapidly emerging as an increasingly significant portion of the dental armamentarium. As advances in laser media and miniaturized delivery systems continue, dental laser research will continue to focus on the risk-benefit ratios and viability of specific laser applications. This paper reviews significant laser research of the past year, covering such areas as laser history, laser types, and laser biophysics. Finally, the future implications for dentistry of the most recent advances in laser therapies on hard and soft tissues is reviewed. PMID- 8401838 TI - Periodontal considerations in prosthodontic treatment. AB - This paper reviews the relationship of periodontics to restorative dentistry. Specific emphasis is placed on the relationship of the restorative margin to the periodontium and the frequent need to alter the placement of the gingival margin so that the restoration will be constructed on sound tooth structure. The prognosis of the sectioned multirooted tooth and other alternatives of treating a shortened dental arch are considered. However, all treatment modalities conclude that a compliant patient and a strong periodontal maintenance program are integral to success, which, with a coordinated effort by the periodontist and the restorative dentist, best assures a successful long-term dental treatment. PMID- 8401839 TI - Dental implants in periodontal care. AB - Dental implants are becoming more commonly used in periodontal offices. The literature related to implants this past year was filled with important material that will change the way we diagnose and formulate treatment plans. This review covers the key points that must be understood to maximize success with dental implants. PMID- 8401840 TI - The possible effect of periodontal diseases on occlusal function. AB - This paper raises new questions about the relationship between occlusion and periodontics. Specifically, it raises questions about the effect of periodontal diseases on mechanoreceptors in the periodontal ligament. Periodontal mechanoreceptors transmit information from the periodontium to various reflexes coordinated by the central nervous system. One of these reflexes is the trigemino neck reflex. Its function is to change the position of the head, neck, and jaws on a moment-to-moment basis, and it powerfully influences the occlusal position. This paper raises questions about the consequences of periodontal diseases on all reflexes that depend on periodontal mechanoreceptors, and specific questions are raised about the effect of periodontal disease on the trigemino-neck reflex because of its extreme importance to the way we analyze and treat occlusion. PMID- 8401841 TI - Outcome studies of periodontal treatment modalities. AB - During the past three decades a significant body of information has been produced in several clinical research centers evaluating the therapeutic responses of the periodontium to different treatment modalities. These studies provide ample evidence that periodontal therapy is effective in controlling periodontal disease, and that dentitions can be preserved for extended periods of time following treatments and proper maintenance. In general, there are no striking differences between the periodontal treatments evaluated. Therefore, the current trend is from a resective to a more conservative, anti-infective therapeutic approach and evaluation of the patient. PMID- 8401842 TI - Bibliography of the current world literature. Periodontology. PMID- 8401843 TI - Periodontal manifestations of systemic disease and management of patients with systemic disease. AB - Many studies show a strong association between diabetes mellitus and risk for periodontal disease destruction. Patients with non-insulin-dependent diabetes mellitus have an increased risk of developing destructive periodontal disease. Under similar plaque conditions, adult patients with long-term, poorly controlled diabetes mellitus have more attachment and bone loss than controlled diabetic patients. Most patients with diabetes mellitus respond to conventional periodontal treatment, but in some cases the response may be related to the degree of metabolic control. Periodontal treatment may have a beneficial effect on the metabolic status of poorly controlled diabetes. Tetracycline therapy may be an effective adjunctive treatment in the management of periodontal disease in diabetic patients by blocking collagenase-dependent periodontal tissue destruction. Pyostomatitis vegetans is frequently associated with chronic inflammatory bowel disease and is a marker for the disease. Plaque control with chlorhexidine gluconate should be preceded by mechanical removal of plaque and calculus in patients with leukemia undergoing chemotherapy. A distinct gingival lesion is associated with Wegener's granulomatosis, a potentially fatal disease that, if detected early, has a favorable prognosis. PMID- 8401844 TI - Current concepts in juvenile periodontitis. AB - Juvenile periodontitis is one of the more debilitating periodontal diseases. The disease entity includes two manifestations of aggressive bone destruction, a localized and a generalized form, which affect adolescents at puberty. Recent studies have defined the prevalence of the disease with regard to locale, gender, and ethnicity. In addition, significant advances have been made in evaluating host response effects in juvenile periodontitis. Studies have identified changes in cell surface receptors on neutrophils, which appear to be related to both receptor density and structure. Various molecular techniques, including restriction fragment length polymorphism, have enabled researchers to identify clonal variants of Actinobacillus actinomycetemcomitans--the microorganism most closely associated with the disease. This may prove to be very useful in identifying virulence factors that have a role in disease initiation and progression. Treatment modalities using debridement, surgery, and anti-infective therapy have been proposed for management of the disease. PMID- 8401845 TI - Current epidemiologic consideration of periodontal disease. AB - Two interesting populations in periodontology have emerged this year. National and international reports have indicated that inflammatory periodontal disease is widespread in adolescents, although most of the cases of disease are considered to be in a reversible stage. Nevertheless, the attachment loss present portends serious future problems. The rapidly growing elderly population presents a unique problem of assessment. Current indices do not adequately evaluate recession. Thus, there is substantial underestimation of periodontal disease in the elderly. Further insight into the interrelationships between systemic diseases and local inflammatory periodontal diseases has been reported. Methods to improve statistical evaluation of site-specific periodontal data and address the shortcomings of currently available periodontal indices have been suggested. Finally, the major disadvantage of indices in treatment planning is the lack of specific treatment goals. Examples of the potentially misleading information that results from this lack were presented and the inclusion of treatment goals with treatment needs was suggested. PMID- 8401846 TI - Periodontal considerations in the patient with HIV. AB - The HIV-associated periodontal diseases present unique challenges to the dental practitioner. The presence and severity of HIV-associated gingivitis, HIV associated periodontitis, and possibly necrotizing ulcerative gingivostomatitis, and other oral lesions may indicate the presence and staging of HIV infection. In general, a similar relationship does not appear to exist between adult periodontitis and HIV staging. There is a wide variation in recent reports on the epidemiology of HIV-associated diseases. These variations point to the need for a standard definition for HIV-associated periodontal diseases using both conventional periodontal evaluation criteria and criteria designed specifically for the characteristics of HIV-associated gingivitis and HIV-associated periodontitis. HIV staging, geographic location of the study, antiviral and antimicrobial therapies, and oral habits may also account for many of these differences. Both the HIV-associated gingivitis and periodontitis lesions have similar microbiologic profiles to adult periodontitis lesions for previously identified periodontopathic bacteria. In addition, these lesions may have a unique opportunistic microflora. The pathogenesis of HIV-associated periodontal diseases may be due to the microflora, the effects of HIV and other viral agents, or alterations in the host response. These factors should be taken into consideration in the treatment and prevention of these HIV-associated periodontal lesions. PMID- 8401847 TI - Periodontal disease progression. AB - The progression of periodontitis has been an area of active research during the past year. Prevalence and disease severity studies indicate that severe periodontitis remains a problem of considerable magnitude. The standard for evaluation of periodontitis progression is sequential attachment and bone-level measurements. A vast number of approaches to identify indicators of periodontal disease activity are under development. These include methods to detect bacterial infection or the breakdown of the periodontium. The number of proposed models of periodontal progression has expanded along with the identification of risk factors at the site, subject, and population levels. No model currently under consideration is predictive of future periodontal disease progression. PMID- 8401848 TI - Automated periodontal probing and recording. AB - Attempts to standardize and improve periodontal probes have resulted in the introduction of automated probing devices as well as recording systems and enhanced methods to measure and analyze periodontal data. Newer technologies include constant-force, electronically linked probes, thermal and mobility probes, and the use of automated speech recognition to gather and record information. The coupling of these systems with computers has increased our ability to analyze periodontal disease parameters and to gain a new understanding of criteria necessary to measure disease activity. PMID- 8401849 TI - The current status of determining periodontal prognosis. AB - Determining the prognosis for patients and individual sites with inflammatory periodontal diseases is difficult using present methods. Traditional approaches for predicting when disease activity will occur have proved inadequate in some cases. New diagnostic tests and updated methods of traditional tests will eventually allow clinicians to improve their skills in determining prognosis; however, most of these tests are not yet proved or economically feasible. At present, if a patient is currently healthy and has no past history of attachment loss, their prognosis is favorable. The probability of periodontal breakdown occurring in these patients before the next regularly scheduled maintenance visit is slim. A history of attachment loss is the best predictor of future loss, but determining when more loss will occur is problematic. Clinical signs of inflammation, especially bleeding on probing and the presence of suppuration, also appear to predict breakdown, but to a lesser degree than previous attachment loss. It appears that the future holds much promise for improving our ability to make accurate prognoses, especially in cases of aggressive periodontal problems, but until these advances are well tested, traditional methods continue to be the standard. PMID- 8401850 TI - The current status and future prospects of altering the pathogenic microflora of periodontal disease. AB - Modern periodontal therapy aims to suppress or eradicate periodontal pathogens and to maintain a posttreatment flora that is compatible with health. Systemic and local antimicrobial therapies have demonstrated effectiveness against several periodontal pathogens. Sustained-release devices, or irrigators, for local application of antimicrobial agents to periodontal pockets show great promise. Replacement therapy and periodontal vaccines, although still in the early research phase, represent very interesting approaches for preventing periodontitis. Using modern molecular genetic methods, it is theoretically possible to clone virulence determinants of periodontal pathogens into a nonpathogenic organism. The carrier organism with the expressed foreign antigens may be implanted in the oral cavity or the intestinal tract to induce long lasting immunity against the targeted periodontal pathogens. PMID- 8401851 TI - Scaling and root planing without overinstrumentation: hand versus power-driven scalers. AB - This review focuses on select current articles chosen for their unique contribution to the literature in scaling and root planing. New studies suggest that thorough root debridement can be achieved without overinstrumentation using certain sonic and ultrasonic scalers. Evaluation of residual plaque and calculus after instrumentation with hand- and power-driven scalers showed sonic and ultrasonic scalers to be equivalent, and in some cases, superior to hand scaling. When modified ultrasonic inserts were compared with unmodified ultrasonic inserts and hand curets, the modified ultrasonic inserts produced smoother roots with the least amount of damage, better access to the bottom of the pocket, better calculus and plaque removal, less operator time, and less operator fatigue than did hand scaling or ultrasonics equipped with unmodified inserts. PMID- 8401852 TI - Mouthrinses in the prevention and treatment of periodontal disease. AB - Although the ideal topical antimicrobial does not exist, some marketed products possess many of the desirable characteristics of such an agent. Antimicrobial agents from a number of different chemical categories are being evaluated continuously. Recent clinical trials evaluating the effectiveness of chlorhexidine, stannous fluoride, prebrushing rinses, phenolic compounds, cetylpyridinium chloride, and new agents such as triclosan are described. Other trials involving special patient groups such as the elderly, mentally handicapped, and orthodontic patients, are discussed. Recent articles on alternative roles for topical antimicrobials are also identified. PMID- 8401853 TI - Evaluation of site-specific chemotherapeutic agents in periodontal treatment. AB - The rapid evolution of chemotherapeutic agents and site-specific treatment of adult periodontitis using controlled delivery systems require critical evaluation by the clinician. At present there are no defined guidelines to assist clinicians in their decision-making process. This paper offers background information and guidelines that can be applied to new and established chemotherapeutic agents and site-specific controlled delivery treatment modalities. The guidelines are not intended to be all inclusive or represent a definitive treatise on the subject. However, it is hoped that the factors addressed will serve as impetus for a discriminating consideration of the problems inherent to the evaluation of new therapeutic products, particularly those intended for use as site-specific controlled delivery modalities in the treatment of adult periodontitis. PMID- 8401854 TI - Periodontal regeneration: a review of flap management. PMID- 8401855 TI - Periodontal regeneration using combined techniques. PMID- 8401856 TI - Root coverage grafting for regeneration and aesthetics. PMID- 8401857 TI - Basic considerations in periodontal wound healing to achieve regeneration. PMID- 8401858 TI - Development of the biological concept of guided tissue regeneration--animal and human studies. PMID- 8401859 TI - Biodegradable barriers and guided tissue regeneration. PMID- 8401860 TI - Clinical applications of guided tissue regeneration: surgical considerations. PMID- 8401861 TI - Periodontal regeneration: root surface demineralization. PMID- 8401862 TI - Polypeptide growth factors and attachment proteins in periodontal wound healing and regeneration. PMID- 8401863 TI - Bone grafts and periodontal regeneration. PMID- 8401864 TI - Factors related to periodontal regeneration. AB - The purpose of this chapter is to set the stage for the more detailed information to follow in the succeeding sections of this work. The reader will be familiarized with the terms and definitions commonly used in periodontal wound healing and the methods used to evaluate the results of periodontal therapy. A discussion of the effects of conventional periodontal therapy will then be followed by a review of biological factors that must be considered if periodontal therapy is to result in regeneration. PMID- 8401865 TI - Synthetic bone grafts in periodontics. PMID- 8401866 TI - [Immune system status under effect of low levels of ionizing radiation:studies within the 10 kilometer zone of accident at Chernobyl nuclear plant]. AB - A study was made of the influence of high radiation contamination within the ten kilometer zone of Chernobyl disaster on the structure and function of the immune system of (DBA x C57B1)F1 and DBA mice. The cumulative radiation doses with respect to gamma-radiation, were 0.024, 0.168 and 0.336 Gy. T-lymphocyte proliferation was shown the be activated with all radiation doses mentioned above but with doses of 0.024 and 0.168 Gy, helper T lymphocytes, and with 0.336 Gy, suppressor N-lymphocytes were primarily activated. So, in the former case, some effector functions of the immunity were activated, and in the latter, inhibited. It is concluded that certain hyperstimulation of the lymphoid cell formation (including the increase in T-suppressors), that is induced by long-term exposure to low-level radiation, triggers the mechanisms of autonomous regulation of the system that suppress the radiation-induced lymphocyte proliferation and inhibit the effector functions of the immunity. PMID- 8401867 TI - [Immunologic methods in epidemiologic monitoring of population, affected by radioactive iodine as a result of the accident at the Chernobyl nuclear plant]. AB - Ultrasound investigations of the thyroid gland and determinations of microsomal antibodies have been performed in persons who lived in the town of Korosten (Zhitomir Region) during the Chernobyl accident. A high correlation has been found between ultrasound and immunological results. The immunological screening of the population suffered from the Chernobyl disaster might be successfully used for the autoimmune thyroiditis detection. These data complete those obtained by the ultrasound tests. PMID- 8401868 TI - [Modification of adrenergic regulation of heart function under effect of radio ecological factors within 10-kilometer zone of the accident at Chernobyl nuclear plant]. AB - Rats kept within the ten-kilometer zone of Chernobyl disaster over a period of 30 days exhibited a decreased heart function response to beta-adrenoreceptor stimulus, reduced density of the receptor structures in cardiomyocytes and their affinity to specific agonists. The number of beta-adrenoreceptors in myocardium cells was restored in 6 months, but their affinity and heart function response to the effect of beta-adrenoagonists remained decreased. No significant changes were observed in regulation influenced indirectly via alpha-adrenoreceptors. PMID- 8401869 TI - [Statistical criterium of pollution heterogeneity in the research of biotic factors of radionuclides migration]. AB - The nature of the radionuclide contamination resulting from Chernobyl disaster is in its spatial and aggregate heterogeneity determined by the interaction of systematic and incidental factors. To study the radionuclide migration in the soil-plant system, an independent statistical index of contamination heterogeneity H = RAsAp is proposed which is calculated as a Brave-Pirsens coefficient of linear correlation between radioactivity of soil and dry biomass of plants grown on it and which allows to determine relative range of contamination that appears in the form of "hot particles". An adequacy of this index is substantiated, and it is used to analyze the experimental results on radionuclide migration. PMID- 8401870 TI - [Changes of radiosensitivity in animals after exposure to radiation within the territory of accident at the Chernobyl nuclear plant]. AB - Mice exposed inside of 10-km zone around Chernobyl Atomic Station with follow-up acute irradiation were studied to find the changes in their life-span. It was found that the results essentially depend on the interval between exposition in zone and acute irradiation and space of time after irradiation. PMID- 8401871 TI - [Radiation exposure of cultured tissue cells and animals (mice) within 10 kilometers zone of accident at Chernobyl nuclear plant. Effect on radiosensitivity to future radiation]. AB - The effect of chronic low-level radiation exposure on radiosensitivity to posterior acute irradiation at high doses has been studied. Cells and mice were exposed within the ten-kilometer zone of Chernobyl disaster during various spaces of time (1-12 days), then over one or more days were additionally irradiated by doses of 1-4 Gy. It was shown that no adaptive response was developed under chronic exposure of cells and mice within the zone of disaster. On the contrary increased sensitivity to posterior irradiation was revealed. The number of cytogenetic damages of cultured tissue cells and marrow cells (chromosome aberrations and micronuclei) increases, the spectrum of aberrations being shifted to chromosome type, cells with multiaberration appearing. The decay of chromatine increases indicating an interphase death; the number of leucocytes in peripheral blood decreases. PMID- 8401872 TI - [Regulation of glucose transport in rat thymocytes after chronic gamma irradiation]. AB - Chronic gamma-irradiation (1 Gy, 100 mR/hr) increases cell titre in rat thymus and decreases glucose transport activation by phorbol ester. Animals kept in the Chernobyl evacuation zone over a period of 30 days exhibit reduced thymus cellularity and decreased PMA (200 nM) stimulation of glucose uptake by thymus cells. PMID- 8401873 TI - [Phenomenon of adaptive resistance to gamma-irradiation of colony-forming units (CFU-S): Manifestation in exogenous test]. AB - Adaptive response of colony forming units in mice spleen (CFU-S) was investigated in exotest after preirradiation with doses of 0.3-5.0 sGy of 60Co. Adaptive radioresistance was manifested from 4 hours till 28 days after each challenge dose. Low dose rate preirradiation is less effective as adaptive response inducing action than high dose rate one. PMID- 8401874 TI - [ Multigeneration carcinogenesis in the evaluation and prognosis of delayed radiation effects]. AB - Radiation multigeneration carcinogenesis' mechanisms are associated with inheritance of non-specific genome lesions. Evidence indicates that the effect is significantly enhanced by some chemical carcinogens action on the offspring of irradiated parents. Hereditary character of the phenomenon is a prerequisite to accumulation of these genetic determinants in the population. The risk of radiation multigeneration carcinogenesis for man could be as much as 0.4 x 10(-2) Sv-1 over one generation. PMID- 8401875 TI - [Effect of low-dose ionizing radiation on radiosensitivity to the next irradiation]. AB - The action of low dose irradiation on the radiosensitivity of HeLa cells was studied. It was shown that preliminary irradiation of sells by dose of 3 cGy induces the adaptive response: decreasing of number of cells with micronuclei after posterior irradiation by doses of 2.0 and 3.0 Gy. The maximum level of the adaptive response is reached in 4 h and is observed in 3 cell cycles. When the dose of preliminary irradiation increases to 40 cGy the adaptive response is not observed, however the radiosensitivity of cells increases. PMID- 8401876 TI - [Effect of ionizing radiation on the thymus stroma of embryos and newborn mice]. AB - The lobes of thymus of newborn mice irradiated on 17 day of embryonic life or on 1 day after the birth were transplanted under the renal capsule of adult mice. The recipients were thymectomized, lethally irradiated and reconstituted by syngeneic bone marrow cells treated by monoclonal anti-Thy-1-antibody and complement 1 month before the transplantation of thymus. Colonization of the thymus transplants, number and functional activity of T cells in peripheral lymphoid organs of recipients were measured 30 day after the transplantation of thymus. The colonization of the thymus transplants was decreased after irradiation of the thymus donors in doses of 6 and 8 Gy. The number of L3T4+ cells (T helpers) in the lymph nodes of the recipients and the level of the humoral immune response on thymus-dependent antigen were decreased after the exposure of the thymus donors to dose of 1 Gy or higher and the level of graft versus-host reactivity of lymph node cells--after the dose of 2 Gy and higher. The degree of the suppression of the humoral immune response was higher when the donors of thymus were irradiated in embryonal period of development than after the birth. Thus the damage of the components of thymic microenvironment responsible for the T cell development can be arisen by relatively low doses of ionizing irradiation. PMID- 8401877 TI - [Dynamics in changes of a1-thymosin level under irradiation of mice thymus in vivo and in vitro: relation to the antibody-antigen production of epithelial thymus reticulum]. AB - In 3- and 9-month experiments with mice, a study was made of the effect of radiation on serum alpha 1-thymosine concentration after whole-body irradiation and local exposure of the thymus at doses of 1-20 Gy. The effect of 137Cs-gamma rays on the in vitro cultured thymus stroma cells, with respect to alpha 1 thymosine secretion, and the influence of local irradiation of the thymus of production of autoantibodies that react with epithelial thymus cells were studied. Both whole-body irradiation and local exposure of the thymus were shown to cause changes in the alpha 1-thymosine content of the blood plasma. The direction and dynamics of the changes observed are different with whole-body and local exposure. Irradiation of cultured thymus cells of mice causes alterations in alpha 1-thymosine secretion, that is, stimulation at a dose of 1 Gy and inhibition at higher doses. With respect to dose- and time-response, these changes are closer to those observed in alpha 1-thymosine concentration in mouse serum after whole-body irradiation than after local exposure of the thymus in vivo. At remote times after local irradiation of the thymus with doses of 1-10 Gy, autoantibodies are found in mouse serum that react with epithelial cells of the thymus stroma. Autoantibodies are absent at doses of 15 and 20 Gy. PMID- 8401878 TI - [Effect of thermal shock on resistance of filamentous Bacillus subtilis strain to some hazardous substances]. AB - When grown at 30 degrees C and heat shocked in a liquid medium at 45 degrees C, the filamentous cells of Bacillus subtilis 168 became more sensitive to subsequent killing with N-methyl-N'-nitro-N-nitrosoguanidine and UV light but not with gamma-rays. Certain characteristics (for instance, the increased tolerance to damaging agents at 30 degrees C and the time-dependent changes in the sensitivity to MNNG induced by thermal shock) evidence against direct involvement of repair systems in this phenomenon. PMID- 8401879 TI - [Radiosensitizing effect of hyperthermia (41.5C, 30 min) in stem cells of mouse spermatogenic epithelium]. AB - Mouse testes preheated up to 41.5 degrees C for 30 min exhibit an increased radiosensitivity of testis epithelium stem cells. High death rate is described by a shift (by approximately 4 Gy) in the "dose-response" curve to the Abscissa left. The DMF value for hyperthermia depends on the cell survival rate, at which calculations are made, and varies from 2.1 to 1.4 within the range under study. PMID- 8401880 TI - [Genetic effect of incorporated 137 Cs in male mice after single injection of the isotope]. AB - A comparative study of the incidence of genetic damages induced in mouse male germ cells by a single injection of 137Cs and external gamma-irradiation shows that the incidence of dominant lethal mutations in the postmeiotic cells is similar with both radiation types. The frequency of reciprocal translocations in stem cell spermatogonia was considerably lower with 137Cs than with external gamma-irradiation which might be attributed to the prolonged effect of the radionuclide. PMID- 8401881 TI - [Genetic effect of incorporated 137 Cs on male mice after long-term administration of the isotope]. AB - A comparative study has been made of the incidence of genetic damages to male mouse germ cells induced by chronic action of incorporated 137Cs and external gamma-radiation. It has been shown that the genetic efficiency was nearly the same with both radiation types under conditions of equal, with respect to time and value, absorbed dose formation in gonads. PMID- 8401882 TI - [Cyclosporine A fails to prevent thymocyte death induced by ionizing radiation and hydrocortisone]. AB - In experiments with albino mongrel mice the influence of cyclosporine A on thymocyte death in vivo under the effect of ionizing radiation and hydrocortisone has been investigated. The effect was assessed by the amount of soluble polydeoxynucleotides (PDN) that are formed in the cells. Dose dependence of the PDN yield and kinetics of their formation in thymocytes as well as electrophoretic mobility of PDN were studied after irradiation and the cyclosporine A treatment. Cyclosporine A was shown to have no effect on the thymocyte death induced by irradiation in vivo. The same results were obtained with hydrocortisone. Possible mechanisms of action of cyclosporine A on death of lymphocyte and proliferating cells are discussed. PMID- 8401883 TI - [Ozone holes: prognosis of human photo-ecology. Protection and sensitization]. AB - Photoecological consequence of ozone holes are considered. Analysis of the total UV-radiation intensity and the short-wave edge of atmospheric transmittance depending on ozone content is presented. Characteristic times of some photodamages to skin and eyes have been estimated under ozone content reducing. A role of photosensitizers including drugs in phototoxical effects is discussed. Possible ways of photoprotection are considered. PMID- 8401884 TI - [Atmospheric nuclear researches and environmental pollution with radioactive substances]. PMID- 8401885 TI - Economic madness. PMID- 8401886 TI - Vitamin K: the debate rumbles on. PMID- 8401887 TI - AIDS project offers inspiration. PMID- 8401888 TI - Nursing and the law. Mother or foetus--whose rights are paramount? PMID- 8401889 TI - Legal restrictions on the use of human tissue in research. PMID- 8401890 TI - Changing perspectives in autism. PMID- 8401891 TI - Aptitude testing for specialization. PMID- 8401892 TI - War burns: a simplified resuscitation protocol. AB - The fluid resuscitation of burn injuries in mass casualty situations, such as war, presents a formidable challenge. The possibility of overwhelming numbers of patients to be treated by inexperienced personnel dictates the adoption of a safe, simple and effective regimen. In the burn treatment protocol presented here, this regimen is computed in advance and displayed in a simple tabular form. PMID- 8401893 TI - Diagnosis of intradural conus and cauda equina tumours. AB - Tumours in the conus medullaris, the cauda equina and filum terminale are rare, and present a varied clinical picture. There is often a delay between onset of symptoms and diagnosis. However, in most cases, a detailed history and clinical examination will indicate further investigation. PMID- 8401894 TI - Depression in the terminally ill. AB - Clinical depression frequently goes unrecognized and undertreated, yet it is eminently treatable. It is not an inevitable consequence of terminal illness in the cognitively intact patient--the majority of patients with advanced malignant disease are not depressed. This review examines the epidemiology, causes and treatment of depression in the terminally ill. PMID- 8401895 TI - Management of learning disability in the general hospital. AB - Learning-disabled people need to use the same medical services as the rest of the population. Once referred to a general hospital their diagnosis and treatment must take into account any special needs generated by their disability. This article describes the way in which this can be achieved and the resource implications. PMID- 8401896 TI - Progressive multifocal leucoencephalopathy. AB - Progressive multifocal leucoencephalopathy (PML) is a progressive, usually fatal, demyelinating disease of the CNS that is associated with infection of oligodendrocytes by the papovavirus JC. The incidence of PML is rising as a consequence of the AIDS epidemic. PMID- 8401897 TI - Hepatitis C: a novel cause of progressive multifocal leucoencephalopathy. PMID- 8401898 TI - Colonic crunch: a new physical sign. PMID- 8401899 TI - Problems with a fractured jaw. PMID- 8401900 TI - A charter for house officers. AB - Recently there have been several initiatives to improve the working conditions and training of junior doctors. However, monitoring is infrequent considering the rapid changes in the health service, thus a mechanism to detect early problems is necessary. With this in mind, we have drawn up a charter for house officers that outlines standards which they should expect and the action they should take if they are not met. PMID- 8401901 TI - Future training of hospital doctors. AB - Many changes are occurring in the organization and management of postgraduate medical education and training. Hospital Doctors: Training for the Future (Department of Health, 1993) highlights the need for shorter training programmes and structured curricula. A number of issues still require analysis, particularly the resource implications of expanding the consultant grade. PMID- 8401902 TI - What the patient is looking for. AB - 'Each patient considers his complaint as one of great peculiarity or perhaps obscurity, demanding prompt attention' (Longcope, 1962). This quote serves to illustrate the importance of examining the consultation from the patient's perspective. It is this problem that is considered in this article, the first in a series of 15. PMID- 8401903 TI - Decision making in surgery: acute anorectal sepsis. AB - Acute perineal inflammation is a common surgical emergency, presenting in most cases with an abscess arising from infection within the many tissue planes in this area. Simple drainage of such an abscess leads to immediate symptomatic relief but this should be accompanied by diagnostic manoeuvres that allow any underlying fistulae to be found and eradicated. PMID- 8401904 TI - Obese patient with coexisting anaesthetic problems. PMID- 8401905 TI - Consent to medical treatment: teenagers and the law. PMID- 8401906 TI - Minimally invasive surgery in gynaecology. PMID- 8401907 TI - Minimally invasive surgery in gynaecology. PMID- 8401908 TI - Minimally invasive surgery in gynaecology. PMID- 8401909 TI - Intravascular ultrasonography. PMID- 8401910 TI - Intravascular ultrasonography. PMID- 8401911 TI - Problems with fetal medicine. PMID- 8401912 TI - Tachykinin receptors mediating responses to sensory nerve stimulation and exogenous tachykinins and analogues in the rabbit isolated iris sphincter. AB - 1. We have used selective tachykinin receptor agonists and antagonists to investigate the nature of the receptors mediating responses to endogenous and exogenous tachykinins in the rabbit iris sphincter preparation in vitro. 2. The NK1-selective agonist, substance P methyl ester, induced contraction with a pD2 of 9.16 indicating the presence of NK1 receptors. In confirmation, the NK1 selective antagonist, GR82334, competitively antagonized responses to substance P methyl ester with high affinity (pKB 7.46). 3. NK3 receptors also mediate contraction since NK3-selective agonists exhibited high potency, e.g. the pD2 of [Me-Phe7]-neurokinin B was 9.67, and their responses were not inhibited by GR82334 (10 microM). 4. NK2 receptor activation does not seem to contribute to contraction since the NK2-selective agonist [beta-Ala8]-neurokinin A(4-10) had relatively low potency (pD2 6.43), and the NK2-selective antagonists MEN10207 (1 microM) and L-659,877 (10 microM) were inactive or had low affinity, respectively. 5. GR82334 (1 microM) significantly inhibited responses to electrical field-stimulation of non-adrenergic non-cholinergic sensory nerves (3, 10 and 30 Hz), and caused a rightward shift of the log concentration-response curve to bradykinin (lateral shift ca. 1000 fold). Higher concentrations of GR82334 (10 microM) significantly attenuated responses to capsaicin (1-60 microM) whilst completely abolishing responses to field-stimulation (3, 10 and 30 Hz) and bradykinin (1 nM- 3 microM). 6. In conclusion, NK1 and NK3 receptor activation results in contraction of the rabbit iris sphincter. The contractile response following sensory nerve stimulation by bradykinin, capsaicin and electrical field stimulation results from NK1 receptor activation. PMID- 8401913 TI - Effects of beraprost on the transmembrane potentials of guinea-pig ventricular muscles during normoxia and hypoxia-reoxygenation. AB - 1. The present study was performed to determine whether beraprost, a new stable analogue of prostacyclin, may exert beneficial effects on the transmembrane action potentials during normoxia and hypoxia-reoxygenation in isolated right ventricular muscles of the guinea-pig. 2. Under normal oxygenation, beraprost (0.01-100 mumol-1) had no effects on the electrophysiological parameters. 3. Hypoxic conditions induced a decrease in action potential duration (APD) without affecting other action potential parameters. Beraprost inhibited this hypoxia induced decrease in APD. However, beraprost had no effect on the decrease in contractile force induced by hypoxia, whereas it significantly improved the recovery of contractile force after reoxygenation. 4. Pinacidil-induced shortening of APD was not antagonized by beraprost. 5. Hypoxia significantly decreased the myocardial adenosine triphosphate (ATP) level, which was also prevented by beraprost. 6. These results suggested that beraprost may inhibit the hypoxia-induced shortening of APD by some mechanisms which contribute to the maintenance of muscle ATP level. PMID- 8401914 TI - Metabolism of endothelin-1 and big endothelin-1 by recombinant neutral endopeptidase EC.3.4.24.11. AB - 1. Inhibitors of neutral endopeptidase EC.3.4.24.11 (NEP) have been shown to attenuate the hypertensive effect of big-endothelin-1 (BET-1) in rats. To determine whether NEP converts BET-1 to endothelin-1 (ET-1), the effect of a recombinant NEP (rNEP) on BET-1 and on ET-1 was assessed in vitro. 2. Incubation of [125I]-ET-1 with 1 microgram ml-1 of rNEP resulted in degradation of the peptide within minutes. Increase in the amount of rNEP to 10 micrograms ml-1 led to total cleavage of [125I]-ET-1 within seconds. 3. Phosphoramidon (10 microM) or SQ-28,603 (100 microM) totally suppressed the degradation of [125I]-ET-1 by rNEP. 4. The degradation of [125I]-BET-1 by either 1 or 10 micrograms ml-1 of rNEP was much slower than that of [125I]-ET-1. Again, both phosphoramidon and SQ 28,603 protected the peptide from degradation. 5. Intact [125I]-ET-1 was not observed when [125I]-BET-1 was incubated with rNEP. 6. These data show that neutral endopeptidase EC.3.4.24.11 is not an endothelin converting enzyme. PMID- 8401915 TI - Mechanism of carbachol-evoked contractions of guinea-pig ileal smooth muscle close to freezing point. AB - 1. The effect of lowering the temperature to near freezing-point upon the contractions and [3H]-inositol phosphate responses to carbachol were investigated in longitudinal smooth muscle from the guinea-pig ileum. 2. The peak amplitude of the contraction to a single application of 100 microM carbachol was the same at 37 degrees C and temperatures near freezing-point. However, the sensitivity to carbachol was reduced upon lowering the temperature and the time to peak contraction was increased from 5-10 s to 2-10 min. Even when the temperature was maintained near freezing-point, washing off carbachol produced a relaxation and eventual return of tension to basal levels. 3. Incubating the tissue in 140 mM K+, calcium-free solution or in calcium channel antagonists significantly reduced the carbachol-induced contraction to 10-30% of the control at 37 degrees C and also at 3 degrees C. Thus the majority of the activator calcium required for contraction entered the tissue via voltage-dependent calcium channels (VDCs) at both 37 degrees C and 3 degrees C. 4. The contractions produced by high potassium solutions were less at temperatures close to freezing-point than those at 37 degrees C suggesting that voltage-dependent calcium entry was inhibited as the temperature was lowered. 5. A small part of the contractile response to 100 microM carbachol was resistant to the removal of extracellular calcium at both 37 degrees C and 3 degrees C and this component was increased under depolarizing conditions. This suggests that the release of stored calcium contributes to a minor degree to contraction at both 37 degrees C and 3 degrees C.6. Although 100 microM carbachol produced a statistically significant rise in several [3H] inositol phosphate isomers at both 37 degrees C and 3 degrees C, the production of [3H]-inositol phosphates was less at 3 degrees C than at 37 degrees C and the increase in their production caused by carbachol was much slower.7. These results suggest that the carbachol-induced contraction at 3 degrees C utilizes both calcium entry through VDCs and calcium release from intracellular stores, as at 37 degrees C. The components of the responses dependent upon intracellular calcium release at 37 degrees C and at temperatures near freezing-point were similar. However, the production of [3H]-inositol phosphates, including the calcium-mobilizing second messenger inositol (1,4,5) trisphosphate (Ins(1,4,5)P3), is reduced at such low temperatures. PMID- 8401916 TI - Calcium modulatory properties of 2,6-dibutylbenzylamine (B25) in rat isolated vas deferens, cardiac and smooth muscle preparations. AB - 1. In rat isolated vas deferens the new compound 2,6-dibutylbenzylamine (B25) evoked a series of repeating rhythmic contractions. Concentration-response curves constructed for this effect were bell-shaped, indicating a biphasic effect for this compound. By contrast, B25 depressed heart contractility without any visible positive inotropic or chronotropic activity. 2. Experiments with tetrodotoxin, reserpine, capsaicin, alpha-adrenoceptor blocking compounds and other agents permit us to exclude a release of neuromediators or a direct stimulation of post synaptic receptors to account for the rhythmic effect of B25 in the rat vas deferens. 3. In the same tissue, the increase in 45Ca2+ uptake, the voltage dependency as well as the dependence of the B25-induced rhythmic activity upon the external calcium concentration indicate a direct activation of voltage sensitive calcium channels (VSCC). 4. Verapamil paradoxically stimulated the rhythmic effect of B25 in the rat vas deferens. La3+ was inactive while nifedipine was a weak inhibitor. By contrast Ni2+ and Mn2+ ions were good inhibitors (IC50 < 10(-4) M), suggesting that a possible opening of T-type VSCC underlies rhythmic effect of B25. 5. In radioligand binding studies competition experiments with [3H]-nitrendipine indicated that only at high concentrations was B25 able to interact with dihydropyridine-sensitive binding sites of heart and vas deferens smooth muscle. 6. B25 (3-30 microM) counteracted the inhibitory effects of omega-conotoxin GVIA in field-stimulated rat vas deferens. PMID- 8401917 TI - Contraction of the rat isolated spleen mediated by adenosine A1 receptor activation. AB - 1. A series of adenosine receptor agonists of varying degrees of selectivity induced concentration-dependent contraction of the rat isolated spleen. With the exception of the response to the selective A2A receptor agonist, 2-[p-(2 carboxyethyl)phenylethylamino]-5'-N- ethylcarboxamidoadenosine (CGS 21680), responses to each ligand were blocked surmountably and to a broadly similar extent by 8-p-sulphophenyltheophylline (10(-5) M). 2. There was a significant correlation between the pEC50 values obtained on the spleen and the binding affinities (pKD; measured with [3H]-NECA) for the A1 receptor of pig striatum (r = 0.98, P < 0.001) but not the A2A receptor (r = 0.14, NS). 3. The antagonist potencies of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) and 9-chloro-2-furyl [1,2,3]triazolo[1,5-C]quinazoline-5-amine (CGS 15943) were measured against the prototype selective A1 receptor agonist, R-N6-phenylisopropyladenosine (R-PIA). The resulting pKB values of 8.67 and 7.70, respectively are consistent with the A1 receptor subtype mediating splenic contraction. 4. The response to R-PIA was unaltered in the presence of a concentration (10(-7) M) of CGS 21680 which is 6 fold its KD concentration at the A2A binding site in pig striatum but below the threshold for causing contraction per se; thus, A2A receptors inhibitory to contraction appear to be absent. 5. The response to R-PIA was resistant to blockade by prazosin (10(-7) M) and by nifedipine (10(-6) M) but partially blocked by indomethacin (10(-6) M). 6. The results show that the rat isolated spleen responds to adenosine receptor agonists with contraction. Both the relative potencies of agonists and the effects of antagonists indicate mediation by the A1 receptor subtype. alpha1-Adrenoceptor activation is not involved in contraction but a role for products of cyclo-oxygenase and calcium from a source not dependent on entry through L-channels is implicated. PMID- 8401918 TI - Modulation of opioid antinociception by CCK at the supraspinal level: evidence of regulatory mechanisms between CCK and enkephalin systems in the control of pain. AB - 1. Much evidence in the literature supports the idea that cholecystokinin (CCK) interacts with opioids in pain mechanisms. In this work, we have investigated the supraspinal interactions between enkephalins and CCK, using the hot plate test in mice. 2. Intracerebroventricular (i.c.v.) administration of BDNL (a mixed CCKA/CCKB agonist) induced dose-dependent antinociceptive responses on both paw lick and jump responses. In contrast, using the same test, the i.c.v. injection of BC 264 (a selective CCKB agonist) induced a hyperalgesic effect, which was restricted to paw licking and occurred only at a high dose of 2.5 nmol. 3. In addition, i.c.v. administration of BDNL potentiated the antinociceptive effects of the mixed inhibitor of enkephalin degrading enzymes, RB 101 and of the mu agonist, DAMGO, while BC 264 reduced these effects. 4. Furthermore, at a dose where it interacts selectively with delta-opioid receptors, the opioid agonist BUBU reversed the hyperalgesic responses of BC 264 (2.5 nmol) but was unable to modify the effects induced by BDNL. 5. Taken together, these results suggest the existence of regulatory mechanisms between CCK and enkephalin systems in the control of pain. These regulatory loops could enhance the antinociceptive effects of morphine allowing the opiate doses used to be reduced and thus, possibly, the side-effects to be minimized. PMID- 8401919 TI - Signal transduction pathways involved in the acute potentiation of NMDA responses by 1S,3R-ACPD in rat hippocampal slices. AB - 1. A grease-gap recording technique has been used to investigate the mechanisms underlying the acute potentiation of N-methyl-D-aspartate (NMDA) responses by aminocyclopentane-1S,3R-dicarboxylic acid (1S,3R-ACPD) in area CA1 of rat hippocampal slices. 2. 1S,3R-ACPD (10 microM), but not 1R,3S- ACPD (10 microM), potentiated submaximal responses to NMDA (dose-ratio of 0.81 +/- 0.02 (mean +/- s.e.mean); n = 55), but not to alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA), in a readily reversible manner. Potentiation also occurred in slices treated with 0.2 microM tetrodotoxin, and in slices perfused with Mg(2+)-free medium. 3. 1S,3R-ACPD-induced potentiation was unaffected by the protein kinase inhibitors K-252b (0.1 microM) and staurosporine (1 microM) and the intracellular Ca2+ store depletor, thapsigargin (10 microM). Coapplication of staurosporine and thapsigargin was also without effect. 4. 1S,3R-ACPD-induced potentiation was unaffected by inhibitors of arachidonic acid formation, bromophenacyl bromide (50 microM) and RG80267 (100 microM). Arachidonic acid (10 50 microM) depressed reversibly NMDA-induced responses. The potentiation was unaffected by 8-bromo cyclic AMP (500 microM) or the PKA inhibitor Rp-adenosine 3,5-cyclic monophosphothioate triethylamine (Rp-cAMPS; 50 microM). 5. 1S,3R-ACPD induced potentiation was abolished in slices perfused with Ca(2+)-free medium. The potentiation was also blocked by phorbol-12,13-diacetate (1 microM), in a staurosporine-sensitive manner. 6. It is concluded that the potentiation of NMDA responses by 1S,3R-ACPD is not mediated by protein kinase A or C, by release of Ca2+ from intracellular stores or by arachidonic acid. It involves a Ca2+ sensitive process and is negatively regulated by protein kinase C. PMID- 8401920 TI - Influence of epithelium on the inhibition of melittin-induced contraction of guinea-pig isolated trachea by the potassium channel opener NIP-121. AB - 1. We have investigated the effect of the potassium channel opener, NIP-121, on contraction elicited by melittin (a phospholipase A2 activator) in epithelium intact and epithelium-denuded trachea isolated from guinea-pigs. The effects of NIP-121 were compared with those of isoprenaline, aminophylline and hydrocortisone. 2. In the presence of the cyclo-oxygenase inhibitor, indomethacin (5 microM), melittin (3 micrograms ml-1) caused time-dependent contraction. The melittin-induced contractile response was significantly augmented by removal of the epithelium and was concentration-dependently and completely inhibited by the phospholipase A2 (PLA2) inhibitor, p-bromophenacyl bromide (BPB 10-100 microM), and the lipoxygenase inhibitor, phenidone (10-300 microM). Similar inhibition of the melittin response by BPB (10 microM) and phenidone (10 microM) was observed in the epithelium-denuded trachea. 3. Concentration-dependent inhibition of the melittin response was induced by preincubation with NIP-121 (0.03 and 0.1 microM), isoprenaline (0.001 and 0.01 microM), aminophylline (30 and 100 microM) and hydrocortisone (100 and 300 microM). The effect of NIP-121 was abolished by glibenclamide (1 microM). 4. The inhibitory effect of NIP-121 on the melittin response was greatly reduced by removing the epithelium while that of the isoprenaline, aminophylline or hydrocortisone was not changed. 5. The inhibition of the melittin response by these drugs was similar to their inhibition of the contraction elicited by a low concentration (3 nM) of leukotriene D4 (LTD4) in the epithelium-intact trachea. Inhibition of the LTD4 response by NIP-121 was not observed in the epithelium-denuded trachea. However, higher concentrations of NIP 121 (0.3 and 1 microM) did inhibit LTD4-induced contractions of epithelium denuded trachea.6. These findings suggest that melittin causes epithelium dependent contraction of the guinea-pig isolated trachea which is mediated by products of lipoxygenase activity. NIP-121 may inhibit the melittin response by activating glibenclamide-sensitive potassium channels, which appear to be epithelium-dependent (an indirect effect of NIP-121 apart from its direct effect on the airway smooth muscle) while isoprenaline, aminophylline and hydrocortisone act directly to relax the trachealis smooth muscle. PMID- 8401921 TI - Effects of cromakalim on the electrical slow wave in the circular muscle of guinea-pig gastric antrum. AB - 1. In circular muscle strips of the antrum of guinea-pig stomach, the effects of cromakalim were studied on mechanical activity and intracellular membrane potential. 2. Cromakalim inhibited mechanical activity at concentrations higher than 1 microM, accompanied by membrane hyperpolarization and a decrease in membrane resistance. The hyperpolarization was markedly potentiated in K(+)-free solution and was still observed in the absence of Na+. 3. Slow wave electrical activity was relatively resistant to cromakalim. Changes in its amplitude and frequency were not consistent but blockade of slow waves was never observed. In many preparations cromakalim induced spike-like potentials at the top of slow waves, or when spike-like potentials already existed they were potentiated. However, mechanical activity was always inhibited. 4. Inhibition by cromakalim of the phasic contractions associated with the slow waves, could not be reversed by increasing the external K+ concentration (12-30 mM). 5. The results suggest that in guinea-pig stomach muscle mechanical suppression by cromakalim does not simply result from membrane hyperpolarization or from inhibition of slow waves. A clear dissociation was found between the mechanical and electrical activities. Slow waves, particularly their frequency, are relatively insensitive to membrane hyperpolarization. PMID- 8401922 TI - Estimation of competitive antagonist affinity from functional inhibition curves using the Gaddum, Schild and Cheng-Prusoff equations. AB - 1. The estimation of antagonist affinity from functional experiments in which the effect of a fixed agonist concentration is reduced by a range of antagonist concentrations ('functional inhibition curves') has been considered from both a theoretical and experimental viewpoint. 2. Theoretical predictions are compared with results obtained from the stimulation of [35S]-GTP gamma S binding by acetylcholine to membranes of Chinese hamster ovary (CHO) cells stably transfected with human m1-m4 muscarinic receptors, and inhibition of the stimulated binding by pirenzepine and AQ-RA 741. 3. The usual procedure of applying the Cheng-Prusoff correction is shown to be theoretically invalid, and predictions are made of the size and distribution of errors associated with this procedure. 4. A different procedure for estimating antagonist affinity, using the principles of dose-ratio analysis and analogous to use of the Gaddum equation, is found to be accurate and theoretically valid. 5. A novel method of analysis allows accurate estimation of both antagonist affinity and Schild slope, by fitting the combined data from an antagonist inhibition curve and an agonist activation curve directly to a form of the Schild equation (derived by Waud) using non-linear regression analysis. 6. It is shown that the conventional Schild analysis can be enhanced by treating part of the data as a family of inhibition curves and including in the Schild plot dose-ratios estimated from the inhibition curves. PMID- 8401923 TI - Pharmacological characterization of acetylcholine-stimulated [35S]-GTP gamma S binding mediated by human muscarinic m1-m4 receptors: antagonist studies. AB - 1. We have used dose-ratio analysis to estimate functionally the affinity constants (pKb) and Schild slope factors of a range of selective or atypical antagonists at human muscarinic m1-m4 receptors. 2. The functional response was the stimulation by acetylcholine of [35S]-GTP gamma S binding to membranes from Chinese hamster ovary (CHO) cells stably expressing individual receptor subtypes. 3. A novel experimental design and analysis was used which allowed the estimation of affinity and Schild slope factor from a single antagonist inhibition curve, and the results were compared with other methods of analysis, both theoretically valid and invalid. 4. In general, the affinity estimates were very similar to previously reported values obtained in binding studies with animal tissues and cloned human receptors and the Schild slope factors were close to unity. 5. These results demonstrate the validity of the assay and provide no evidence for species differences in antagonist affinity for muscarinic receptor subtypes. 6. The results confirm both the utility of himbacine in distinguishing between m1 and m4 receptors and a previously reported modest m4-selectivity for tropicamide and secoverine. 7. The cholinesterase inhibitor, tacrine (THA), had a potency profile similar to that of gallamine but with less selectivity. Its affinity could not be determined since it had Schild slope factors of about 2 at all subtypes. 8. o Methoxy-sila-hexocyclium had only a modest selectivity for the m1 subtype. PMID- 8401924 TI - The effect of endothelins on nitric oxide and prostacyclin production from human umbilical vein, porcine aorta and bovine carotid artery endothelial cells in culture. AB - 1. This study has investigated the effects of the endothelin isopeptides, endothelin-1 (ET-1), ET-2 and ET-3 on the production of the endothelium-derived relaxing factors, nitric oxide (NO) and prostacyclin (PGI2) from primary cultures of endothelial cells obtained from human umbilical vein (HUVECS), porcine aorta (PAECS) and bovine carotid artery (BCAECS). 2. NO generation was assessed indirectly by measuring production of cyclic GMP and PGI2 formation was measured by radioimmunoassay of 6-keto PGF1 alpha. 3. In HUVECS, histamine (1 microM) increased cyclic GMP and 6-keto PGF1 alpha production by 12.6 +/- 2.0 and 4.9 +/- 0.7 fold respectively over the corresponding basal values. Haemoglobin (10 microM) and the NO synthase inhibitor NG-monomethyl-L-arginine (10 microM) significantly inhibited the increase in cyclic GMP formation in response to histamine but had no effect on 6-keto PGF1 alpha production. In contrast to histamine, the endothelin isopeptides (ET-1, ET-2 and ET-3; 0.01-1000 nM) produced no significant change in either cyclic GMP or 6-keto PGF1 alpha production in HUVECS. 4. In a separate series of experiments, ET-3 (0.01-1000 nM) also failed to produce any significant change in cyclic GMP or 6-keto PGF1 alpha production from primary cultures of PAECS and BCAECS. In contrast, bradykinin (0.1 microM) and sodium nitroprusside (1 mM) were used as positive control agents and increased cyclic GMP production in these cells. 5. In conclusion, the endothelin isopeptides do not release NO and PGI2 from primary cultures of HUVECS, PAECS and BCAECS. This suggests that endothelin receptors are either absent from these cells or are not coupled to NO or PGI2 production. PMID- 8401925 TI - Comparison of the airways relaxant and hypotensive potencies of the potassium channel activators BRL 55834 and levcromakalim (BRL 38227) in vivo in guinea-pigs and rats. AB - 1. BRL 55834, a novel potassium channel activator, has been compared with levcromakalim (BRL 38227) for its relaxant effects in vivo on the airways and vasculature of the guinea-pig and rat. 2. When administered intravenously 2 min prior to challenge, BRL 55834 and levcromakalim each inhibited histamine-induced increases in airways resistance (Raw) in the anaesthetized guinea-pig, with BRL 55834 showing a 4.5 fold greater potency than levcromakalim (ED25 = 2.5 micrograms kg-1 and 11.3 micrograms kg-1 respectively). By contrast, both compounds had similar hypotensive potencies (ED18 = 8.5 micrograms kg-1 and 6.5 micrograms kg-1 respectively). 3. In the same guinea-pig model, intraduodenally administered BRL 55834 (100 and 250 micrograms kg-1) and levcromakalim (500 micrograms kg-1) each protected against histamine-induced changes in Raw and dynamic lung compliance (Cdyn), both compounds showing a rapid onset of action that persisted for more than 50 min. The lower dose of BRL 55834 had a similar bronchodilator effect to that of levcromakalim, yet both doses of BRL 55834 elicited substantially smaller effects than levcromakalim on mean arterial blood pressure. 4. In the anaesthetized rat, BRL 55834 and levcromakalim each evoked a dose-related inhibition of inhaled methacholine-induced changes in Raw and Cdyn when given i.v., with BRL 55834 showing some four fold greater potency than levcromakalim (BRL 55834: Raw ED35 = 3.7 micrograms kg-1, Cdyn ED35 = 5.9 micrograms kg-1; levcromakalim: Raw ED35 = 16 micrograms kg-1, Cdyn ED35 = 23.5 micrograms kg-1). As in the guinea-pig,BRL 55834 had a reduced propensity to lower mean arterial blood pressure (ED11 = 8 microg kg-1 for BRL55834, 11 +/- 3% being its maximum effect; ED11= 16 microg kg-1, maximum effect= 34 +/- 6% for levcromakalim.5. When administered intraduodenally to anaesthetized rats, BRL 55834 (10, 20 and 100 microg kg-1)evoked rapid and dose-related inhibitions of methacholine-induced Raw and Cdyn changes which persisted for over 30 min. At the lower and middle dose there was little effect on mean arterial blood pressure(<10% fall). Levcromakalim (500 microg kg-1) by contrast elicited transient airways responses that diminished rapidly after 5 min, while the effects on blood pressure were well maintained (>20% at 65 min). Levcromakalim (100 microg kg-1) did not affect airways responses but also evoked a marked and sustained fall in blood pressure.6. BRL 55834, administered per os, prolonged the time to histamine-induced dyspnoea in conscious guinea-pigs. The greatest effect of BRL 55834 was observed when it was administered 60 min prior to challenge, a dose of 0.20 mg kg-1 doubling the mean time to collapse. A similar level of protection was afforded by levcromakalim (1.25 mg kg-1), with maximal activity occurring between 30 and 60 min.7. The present studies in guinea-pigs and rats indicate that BRL 55834 is the first potassium channel activator to exhibit greater bronchodilator potency than levcromakalim but reduced tendency to lower arterial blood pressure. It is suggested that BRL 55834 may have greater potential than levcromakalim as a bronchodilator for therapeutic use in man. PMID- 8401926 TI - Bicuculline-resistant paired-pulse inhibition in the rat hippocampal slice. AB - 1. An initial observation that paired-pulse inhibition in hippocampal slices was increased rather than decreased by bicuculline prompted the present study to explore the mechanism underlying bicuculline-resistant inhibition. 2. In the presence of bicuculline, paired-pulse interactions were dependent on the interpulse interval (i.p.i.) but a medium-latency inhibition was consistently observed at an i.p.i. of 300 to 500 ms. 3. The medium-latency (300 ms) bicuculline-resistant inhibition produced by paired orthodromic stimuli was substantially reduced by 2-hydroxysaclofen and was probably largely mediated by GABAB-receptor activation. Paired-pulse inhibition produced by an orthodromic/antidromic stimulation sequence was not affected by 2-hydroxysaclofen suggesting the possibility that the GABAB-receptors involved in orthodromic inhibition may be located presynaptically on the Schaffer collateral terminals rather than on the postsynaptic surface. The medium latency inhibition was also reduced by baclofen and under some conditions, by adenosine. 4. In addition to the GABAB-component, a hydroxysaclofen-resistant depression of postsynaptic excitability contributed to bicuculline-resistant paired-pulse inhibition at the 300 ms latency. PMID- 8401927 TI - A novel metabotropic glutamate receptor agonist: marked depression of monosynaptic excitation in the newborn rat isolated spinal cord. AB - 1. Neuropharmacological actions of a novel metabotropic glutamate receptor agonist, (2S,1'R,2'R,3'R)-2(2,3-dicarboxycyclopropyl)glycine (DCG-IV), were examined in the isolated spinal cord of the newborn rat, and compared with those of the established agonists of (2S,1'S,2'S)-2-(carboxycyclopropyl)glycine (L-CCG I) or (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid ((1S,3R)-ACPD). 2. At concentrations higher than 10 microM, DCG-IV caused a depolarization which was completely blocked by selective N-methyl-D-aspartate (NMDA) antagonists. The depolarization was pharmacologically quite different from that caused by L-CCG-I and (1S,3R)-ACPD. 3. DCG-IV reduced the monosynaptic excitation of motoneurones rather than polysynaptic discharges in the nanomolar range without causing postsynaptic depolarization of motoneurones. DCG-IV was more effective than L-CCG I, (1S,3R)-ACPD or L-2-amino-4-phosphonobutanoic acid (L-AP4) in reducing the monosynaptic excitation of motoneurones. 4. DCG-IV (30 nM-1 microM) did not depress the depolarization induced by known excitatory amino acids in the newborn rat motoneurones, but depressed the baseline fluctuation of the potential derived from ventral roots. Therefore, DCG-IV seems to reduce preferentially transmitter release from primary afferent nerve terminals. 5. Depression of monosynaptic excitation caused by DCG-IV was not affected by any known pharmacological agents, including 2-amino-3-phosphonopropanoic acid (AP3), diazepam, 2-hydroxysaclofen, picrotoxin and strychnine. 6. DCG-IV has the potential of providing further useful information on the physiological function of metabotropic glutamate receptors. PMID- 8401928 TI - Halothane inhibits the pressor effect of diphenyleneiodonium. AB - 1. We have recently found that diphenyleneiodonium (DPI), a novel inhibitor of nitric oxide (NO) synthase, causes pressor and tachycardic responses in pentobarbitone- but not halothane-anaesthetized rats. The present study investigated the mechanism by which halothane suppresses the pressor response of DPI. The effects of halothane on the pressor response of DPI were also compared with those of other anaesthetic agents. 2. In conscious rats, i.v. bolus injections of DPI (0.025- 1.6 mg kg-1) caused dose-dependent increases in mean arterial pressure (MAP), with ED90 of 0.07 +/- 0.01 mg kg-1 and maximal rise of MAP (Emax) of 59 +/- 2 mmHg. While ketamine potentiated Emax without altering the ED50 and pentobarbitone increased the ED50 without changing Emax of the pressor response to DPI, chloralose, urethane and ethanol displaced the curve to the right and potentiated Emax. In contrast, halothane (0.5-1.25%) dose-dependently and non-competitively reduced the pressor responses to DPI. 3. Intravenous bolus injection of a single dose of DPI (1.6 mg kg-1) caused immediate and large increases in plasma noradrenaline and adrenaline, as well as MAP in conscious rats. Halothane (1.25%) almost completely inhibited these increases. 4. The results suggest that DPI causes a pressor response in conscious rats by activating the sympathetic nervous system and halothane abolishes this pressor response by inhibiting activities of the sympathetic nervous system. The results also show that influences of anaesthetics must be taken into consideration when evaluating pressor response of vasoactive agents. PMID- 8401929 TI - Characterization of the 5-hydroxytryptamine receptor mediating the positive inotropic response in guinea-pig isolated left atria. AB - 1. 5-Hydroxytryptamine (5-HT), in the presence of propranolol (1 microM), atropine (3 microM) and ketanserin (1 microM), induced a positive inotropic response of guinea-pig isolated electrically paced left atria (pEC50 = 7.52). The positive inotropic response was mimicked by alpha-methyl-5-HT (pEC50 = 7.26) and 5-carboxamidotryptamine (5-CT; pEC50 = 6.56) but not by sumatriptan or 1-(m chlorophenyl) piperazine (m-CPP). 2. The 5-HT induced positive inotropic response was competitively antagonized by both mesulergine (pA2 = 7.68) and methiothepin (pA2 = 6.67). Methysergide was a surmountable antagonist at 3 nM producing a rightward shift in the 5-HT concentration-response curve giving an apparent pA2 = 9.2 with no significant reduction in the maximum. At higher concentrations, methysergide behaved as an insurmountable antagonist, significantly reducing the maximum response to 5-HT as well as producing rightward shifts in the 5-HT concentration-response curves. 3. The 5-HT-induced positive inotropic response was not antagonized by either tropisetron (10 microM) or yohimbine (10 microM). 4. The guinea-pig atrial 5-HT receptor does not satisfy the criteria for any of the currently recognised 5-HT receptor subtypes and appears to have some similarities to the atypical 5-HT receptors previously described in other peripheral tissues. PMID- 8401930 TI - Selective reduction of N-methyl-D-aspartate-evoked responses by 1,3-di(2 tolyl)guanidine in mouse and rat cultured hippocampal pyramidal neurones. AB - 1. The effects of 1,3-di(2-tolyl)guanidine (DTG) were examined on the responses of cultured hippocampal neurones to the excitatory amino acid analogues N-methyl D-aspartate (NMDA), kainate, quisqualate and (RS)-alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA). 2. In rat hippocampal neurones loaded with the Ca(2+)-sensitive dye Fura-2, DTG (10-100 microM) produced a concentration dependent depression of the NMDA-evoked rises in intracellular free calcium ([Ca2+]i), an effect that was not modified by changes in the extracellular glycine concentration. DTG (at 50 and 100 microM) also attenuated, although to a lesser extent, the rises in [Ca2+]i evoked by naturally-derived quisqualate. In contrast, 50 and 100 microM DTG did not depress responses evoked by kainate, AMPA and synthetic, glutamate-free (+)-quisqualate although on occasions DTG enhanced kainate- and AMPA-evoked rises in [Ca2+]i. 3. DTG attenuated NMDA-evoked currents recorded from mouse hippocampal neurones under whole-cell voltage-clamp with an IC50 (mean +/- s.e. mean) of 37 +/- 5 microM at a holding potential of -60 mV. The DTG block of NMDA-evoked responses was not competitive in nature and was not dependent on the extracellular glycine or spermine concentration. The block did, however, exhibit both voltage-, and use-, dependency. The steady-state current evoked by naturally-derived quisqualate was also attenuated by DTG whereas those evoked by kainate and AMPA were not. 4. We conclude that DTG, applied at micromolar concentrations, is a selective NMDA antagonist in cultured hippocampal neurones, the block exhibiting both Mg(2+)- and phencyclidine-like characteristics. Given the nanomolar affinity of DTG for sigma binding sites it is unlikely that the antagonism observed here is mediated by sigma-receptors, but the data emphasize the potential danger of ascribing the functional consequences of DTG administration solely to sigma receptor-mediated events. PMID- 8401931 TI - [3H]-5-carboxamidotryptamine labels 5-HT1D binding sites in bovine substantia nigra. AB - 1. [3H]-5-hydroxytryptamine (5-HT) has been shown to radiolabel at least five types of 5-HT binding sites in mammalian brain tissue, 5-HT1A, 5-HT1C, 5-HT1D and 5-HT1D and 5-HT1E (Frazer et al., 1990). Selective masking of 5-HT1A and 5-HT1C receptors, has uncovered binding sites which display both high (5-HT1D) and low (5-HT1E) affinity for 5-carboxamidotryptamine (5-CT). By utilizing [3H]-5-CT we have eliminated a portion of the complex binding (5-HT1E) seen when [3H]-5-HT is used as a radioligand. 2. [3H]-5-CT binding to 5-HT1D sites in bovine substantia nigra was rapid, reversible and saturable, displaying high affinity (Kd = 0.38 +/ 0.04 nM) and low non-specific binding (> 90% specific binding). 3. In bovine substantia nigra, [3H]-5-CT labelled an equivalent number of binding sites to [3H]-5-CT (403 +/- 18 and 362 +/- 20 fmol mg-1 protein, respectively) and binding was sensitive to guanine nucleotides. 4. A linear correlation (r2 = 0.99) existed between the potency of compounds to displace [3H]-5-HT and [3H]-5-CT in bovine substantia nigra. 5. Therefore, [3H]-5-CT is a novel radioligand for the examination of 5-HT1-like binding sites, which under proper experimental conditions can be used to radiolabel selectively 5-HT-1D-like binding sites. PMID- 8401932 TI - Effects of NG-nitro-L-arginine methyl ester on the cardiovascular system of the anaesthetized rabbit and on the cardiovascular response to thyrotropin-releasing hormone. AB - 1. The effects of 300 mg kg-1 of the nitric oxide (NO) inhibitor NG-nitro-L arginine methyl ester (L-NAME) on the regional blood flow, on the flow response to 1 mg kg-1 of thyrotropin-releasing hormone (TRH) and on cerebral blood flow autoregulation were studied in urethane anesthetized rabbits subjected to unilateral sectioning of the cervical sympathetic claim. The blood flow measurements were performed by the tracer microspheres method. 2. The cerebral arteriovenous difference in oxygen saturation (CAVOD) was measured before and after the administration of L-NAME and TRH in order to ascertain whether the effects on cerebral blood flow that were observed were secondary to changes in cerebral metabolism. 3. L-NAME caused a significant decrease in blood flow in several cerebral regions; CBFtot decreased to 72 +/- 4% of control (P < 0.001). An increase in blood pressure and a concurrent decrease in heart rate and cardiac output were noted. 4. In the eye, L-NAME caused a reduction in uveal blood flow which was more pronounced on the sympathetically intact side; in the retina the blood flow decreased to 50% of control on both sides. 5. The administration of TRH in animals pretreated with L-NAME caused a significant increase in blood pressure and cerebral blood flow. 6. In L-NAME-treated animals the CBF was not affected when the mean arterial blood pressure was increased by ligation of the abdominal aorta. 7. The CAVOD increased from 56.0 +/- 5.2 to 73.6 +/- 3.5%, 20 min after the administration of L-NAME. In animals given 1 mg kg-1 TRH after L NAME the CAVOD decreased to 54.6 +/- 4.6%, 5 min after the injection of TRH.8. The results of the present study indicate that endogenous NO is involved in the regulation of regional blood flow and blood pressure in the anaesthetized rabbit. The reduction in cerebral blood flow that was caused by L-NAME was not due to a reduction in cerebral metabolism. An interaction between the NO synthesis/release/effect and the sympathetic nervous system was found in the uvea. There was no evidence for a major involvement of NO in the cardiovascular responses to TRH and autoregulation of cerebral blood flow was not abolished by L NAME. PMID- 8401933 TI - Inhibition of the ATP-sensitive potassium channel by class I antiarrhythmic agent, cibenzoline, in rat pancreatic beta-cells. AB - 1. Cibenzoline, a class I antiarrhythmic agent, was investigated for its effect on the ATP-sensitive K+ channel of pancreatic beta-cells by the patch clamp technique. 2. In perforated patch clamp experiments, cibenzoline depolarized the membrane of single beta-cells and thereafter, caused firing of action potentials in the presence of 2.8 mM glucose. 3. Cibenzoline inhibited the activity of the ATP-sensitive K+ channel in cell-attached recordings in the presence of 2.8 mM glucose and evoked repetitive fluctuations of the baseline current, apparently reflecting the action potentials of the beta-cell. 4. In whole-cell clamp experiments, time-independent outward current was induced by depleting cytoplasmic ATP with 0.1 mM ATP and 0.1 mM ADP in the solution contained in the pipette. The outward current was inhibited by cibenzoline in a dose-dependent manner in the concentration range of 1 microM to 100 microM and half maximum inhibition occurred at 1.5 microM. 5. Cibenzoline blocked substantially the ATP sensitive K+ channel current when applied at the inner side of the membrane in isolated inside-out membrane patches. 6. It is concluded that cibenzoline blocks the ATP-sensitive K+ channel of pancreatic beta-cells and, thereby, stimulates insulin secretion at sub-stimulatory levels of glucose. PMID- 8401934 TI - Induction by endogenous noradrenaline of an alpha 1-adrenoceptor-mediated positive inotropic effect in rabbit papillary muscles. AB - 1. The possible involvement of alpha 1-adrenoceptors in the inotropic and electrophysiological responses to endogenous noradrenaline released by tyramine was examined in rabbit papillary muscles. 2. A concentration-dependent positive inotropic effect was produced by tyramine. This effect of tyramine was not observed in muscles from rabbits pretreated with reserpine. 3. The positive inotropic effect of tyramine was greatly inhibited by propranolol, but not altered by prazosin. However, when beta-adrenoceptors were blocked by pretreatment with propranolol, tyramine still produced a positive inotropic effect, an effect which was antagonized by prazosin. 4. Tyramine caused a decrease in action potential duration (APD) and an increase in action potential amplitude in a concentration-dependent manner. Isoprenaline also produced the same electrophysiological effects. These electrophysiological effects of both agents were inhibited by propranolol. 5. When beta-adrenoceptors were blocked by propranolol, the observed prazosin-sensitive positive inotropic effect of tyramine was not accompanied by any change in APD. In contrast, APD was markedly prolonged by alpha 1-adrenoceptor stimulation with phenylephrine in the presence of propranolol, in association with the positive inotropic effect. 6. It is concluded that in rabbit papillary muscles, endogenous noradrenaline causes a positive inotropic effect predominantly mediated by beta-adrenoceptors, but can still evoke a positive inotropic effect through alpha 1-adrenoceptors when beta adrenoceptor stimulation is eliminated. This suggests that the alpha 1 adrenoceptor-mediated positive intropic mechanism(s) may be masked by simultaneous activation of beta-adrenoceptors. In addition, this study indicates that APD prolongation is not involved in the alpha 1-adrenoceptor-mediated inotropic responses to endogenous noradrenaline. PMID- 8401935 TI - Differential inhibition of a transient K+ current by chlorpromazine and 4 aminopyridine in neurones of the rat dorsal root ganglia. AB - 1. K+ currents were recorded from neurones of the newborn rat cultured dorsal root ganglia, by a whole cell variation of the patch-clamp technique. 2. Chlorpromazine (CPZ), a neuroleptic, reversibly reduced the amplitude of the transient K+ current (referred to as 'IT' hereafter) with a dissociation constant (Kd) of 4.5 microM. The inhibition of the delayed rectifier K+ current (IDR) was much less potent (Kd, 120 microM). CPZ (100 microM) had no effect on the inward rectifier K+ current. 3. The blocking action of CPZ on IT was about seven times more potent than that of 4-aminopyridine (4-AP) which had a Kd of 31 microM. The inhibition of IT followed one-to-one binding stoichiometry with both drugs. 4. The decay time course of IT was not affected by CPZ, whereas 4-AP markedly accelerated the decay phase of IT. 5. The steady-state inactivation curve of IT was shifted in the negative direction (about 5 mV) by CPZ, whereas the curve was shifted in the positive direction (about 13 mV) by 4-AP. 6. The recovery from inactivation as measured by a conventional double pulse protocol was described by two exponential components in the control solution. CPZ markedly reduced the first component and slowed down the recovery from inactivation. In contrast, in the presence of 4-AP, the peak amplitude of IT was rather increased by a preceding IT possibly through voltage-dependent unbinding of 4-AP molecules. 7. These results indicate that CPZ has a preferential blocking action on IT and the mechanism underlying this block is markedly different from the mechanism underlying the blocking action of 4-AP. PMID- 8401936 TI - Effects of heparin on the vasodilator action of protamine in the rabbit mesenteric artery. AB - 1. The effects of protamine on the rabbit isolated small mesenteric artery were investigated both in the presence and in the absence of heparin, by the isometric tension-recording method. 2. The dissociation constant for the binding of heparin to protamine has never been previously reported, so in order to minimize the effects of protamine, known to have a vasodilator action, and to examine only the effects of a heparin-protamine complex, the experiments with heparin were performed in the presence of high concentrations of heparin (21-700 u ml-1), concentrations at which heparin itself does not affect the vascular tone. 3. Protamine (15-500 micrograms ml-1), in the absence of heparin, was found to inhibit (P < 0.05) noradrenaline (1 microM)-induced contractions both in endothelium-intact and in endothelium-denuded tissues. 4. Such vasodilator action of protamine in either endothelium-intact or -denuded tissues continued, even in the presence of excess heparin at a heparin/protamine (H/P) ratio of 1.4 u micrograms -1, but was almost completely blocked in the presence of a much greater excess of heparin (H/P ratio > or = 4.7 u micrograms -1): heparin was present both before and during the application of protamine. 5. The vasodilator action of protamine in the absence of heparin was prolonged both in the endothelium-intact and -denuded tissues after protamine had been washed out from the bath with Krebs solution. Although this washing out with a Krebs solution containing excess heparin (4.7 u ml-1) readily reversed such prolonged vasodilator action of protamine both in the endothelium-denuded strips and in the endothelium-intact strips which had been pretreated with inhibitors of the endothelium-derived relaxing factor (EDRF) pathway, it did not affect the prolonged vasodilator action of protamine in the endothelium-intact strips which received no pharmacological intervention.6. These results suggest that: (1) only protamine, not a heparin-protamine complex, exerts vasodilator action in vitro; (2) the vasodilator action of protamine presumably has an EDRF-mediated component;and (3) protamine probably exerts its direct vasodilator action without entering the smooth muscle cell. PMID- 8401937 TI - Protection by nicorandil against the dysfunction of the central vagal baroreflex system following transient global cerebral ischaemia in dogs. AB - 1. A possible cerebroprotective effect of nicorandil was investigated in a canine model of 5 min global cerebral ischaemia, and compared with protective effects of nitroglycerin and nicardipine. 2. Cerebral ischaemia was produced by occlusion of the left subclavian and the brachiocephalic arteries with preceding ligation of the intercostal arteries. The decrease in baroreflex sensitivity (BRS), measured by phenylephrine-induced reflex bradycardia, was used to assess the cerebroprotective effect. 3. Nicorandil (10 or 30 micrograms kg-1 min-1, i.v.), nitroglycerin (3 micrograms kg-1 min-1, i.v.) or nicardipine (0.3 micrograms kg-1 min-1, i.v.) were infused for 60 min just before ischaemia. Nitroglycerin and nicardipine decreased mean arterial blood pressure to an extent similar to that induced by the lower dose of nicorandil. Blood flow in the dorsal medulla oblongata was increased by nicorandil and nicardipine, but not by nitroglycerin. 4. Nicorandil at both doses and nitroglycerin prevented the post-ischaemic decrease in BRS. In these cases, bilateral vagotomy during the reperfusion period decreased BRS, indicating that the vagal component of BRS was protected from ischaemia. On the other hand, nicardipine failed to exert a cerebroprotective effect. 5. The present study suggests that nicorandil may possess a direct cerebroprotective effect and that its property as a nitrate might, at least in part, be important for the observed cerebral protection. PMID- 8401938 TI - Blockade of adenosine receptors unmasks a stimulatory effect of ATP on cardiac contractility. AB - 1. The effects of ATP, alpha,beta-methylene ATP and beta,gamma-methylene ATP on the contractile tension of guinea-pig isolated left atria were evaluated. 2. ATP (1-100 microM) produced a concentration-dependent negative inotropic effect; this response was converted to a positive inotropic effect in the presence of the antagonist of adenosine A1 receptors, 1,3-dipropyl-8-cyclopentylxanthine (DPCPX; 0.1 microM), and in the presence of 8-phenyltheophylline (10 microM), an antagonist of A1 and A2 receptors. 3. The positive inotropic effect of ATP was antagonized by the P2 receptor antagonist, suramin (500 microM). Reactive blue 2 (30-500 microM), a putative P2y receptor antagonist, concentration-dependently reduced and finally abolished the effect of ATP. 4. In the presence of 8 phenyltheophylline, the stable analogues of ATP, alpha,beta-methylene ATP and beta,gamma-methylene ATP (1-30 microM), produced a concentration-dependent increase in atrial contractility of a lesser degree than that induced by ATP. 5. The results suggest that when inhibitory adenosine receptors are blocked, ATP produces a positive inotropic effect, probably mediated by P2y receptor stimulation. PMID- 8401939 TI - Altered cardiac adrenergic neurotransmission in streptozotocin-induced diabetic rats. AB - 1. Functional alterations of the sympathetic neuroeffector junction of the left atria were studied in rats with streptozotocin-induced diabetes. 2. Eight to 12 weeks of diabetes resulted in a marked decrease in the positive inotropic response of left atria to electrical field stimulation (EFS). 3. The overflow of [3H]-noradrenaline from diabetic left atria caused by EFS was much less than that from control preparations. 4. The concentration-response curves showed no change in sensitivities of the left atria to exogenous noradrenaline and tyramine in diabetic rats. The maximum positive inotropic response to these agents were similar in diabetic and control animals. 5. The left atrial content of noradrenaline was not significantly changed in diabetic rats. The cocaine sensitive uptake of [3H]-noradrenaline was also unaltered. 6. Atropine enhanced the positive inotropic response and [3H]-noradrenaline overflow induced by EFS in control left atria. Similarly, yohimbine caused an enhancement of EFS-evoked inotropic response in control atria. However, these effects of the antagonists were not observed in diabetic left atria. 7. It is concluded that the decrease in the positive inotropic response of the left atria to EFS in diabetic rats is caused by an impairment of noradrenaline release from the sympathetic nerve terminals through a calcium-dependent exocytotic mechanism. The present results also indicate that presynaptic alpha 2-adrenoceptors and muscarinic receptors that are linked to inhibition of the noradrenaline release during nerve stimulation may be functionally impaired in diabetic animals. PMID- 8401940 TI - Caffeine-induced decreases in the inward rectifier potassium and the inward calcium currents in rat ventricular myocytes. AB - The effects of high (20 mM) concentrations of caffeine were studied on the transmembrane voltage and currents in rat single ventricular myocytes by the whole cell configuration of the patch clamp technique. Rapid application of caffeine released Ca2+ from the sarcoplasmic reticulum and induced a Ni(2+) sensitive transient inward current with concomitant change of the transmembrane voltage from -72.6 +/- 0.4 to -68.0 +/- 0.6 mV (n = 4). Maintained application of caffeine lengthened the action potential duration (APD90) from 66.7 +/- 16.9 to 135.1 +/- 34.1 ms (n = 4) and depressed the amplitude of both the inward rectifier potassium and the inward calcium currents. It is concluded that these effects of caffeine should be recognized when it is used as a tool to study electromechanical coupling. PMID- 8401941 TI - Different effects of aspirin on blood pressure of spontaneously hypertensive rats (SHR) with high and spontaneously low levels of blood pressure. AB - Spontaneously hypertensive rats (SHR) of the Okamoto strain with blood pressure above 161 mmHg and SHR with blood pressure levels of less than 160 mmHg were treated with oral doses of aspirin (100 mg kg-1) for three days. Whereas the blood pressure of SHR with blood pressure above 161 mmHg was decreased by aspirin, the blood pressure of SHR below 160 mmHg was increased by aspirin. The extent and direction of blood pressure change by aspirin was strongly correlated with the blood pressure of SHR before treatment (r = -0.88). The effect of aspirin supports an important role for endogenous prostanoids in the regulation of blood pressure of SHR. PMID- 8401942 TI - Antagonism of the inhibitory action of neuropeptide Y (NPY) in guinea-pig trachea by the C-terminal fragment NPY (2-36). AB - The C-terminal fragment of neuropeptide Y (NPY), NPY(2-36) was used as a means of discriminating between two differently located NPY receptor sites in guinea-pig trachea. Both NPY and NPY(2-36) reduced the maximal relaxation elicited by vasoactive intestinal peptide (VIP). In contrast, the C-terminal fragment did not mimic the inhibitory action of NPY on the noradrenaline-(NA) evoked response. However, pretreatment of the trachea with 30 nM NPY(2-36), 5 min before generating NA and VIP concentration-response curves in the presence of NPY, abolished the inhibitory effect of NPY on NA-elicited response but did not affect the modulatory action of NPY on VIP-induced relaxation. These results suggest that the two differently located NPY receptor sites in guinea-pig trachea are of two distinct subpopulations. PMID- 8401943 TI - Inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate binding sites in smooth muscle. AB - 1. We have previously demonstrated that activation of M3 muscarinic receptors increases inositol 1,4,5-trisphosphate (InsP3) and inositol 1,3,4,5 tetrakisphosphate (InsP4) accumulation in colonic smooth muscle. 2. In the present study, we demonstrate the existence of InsP3 and InsP4 binding sites in colonic circular smooth muscle by use of radioligand binding methods. Both [3H] InsP3 and [3H]-InsP4 bound rapidly and reversibly to a single class of saturable sites in detergent-solubilized colonic membranes with affinities of 5.04 +/- 1.03 nM and 3.41 +/- 0.78 nM, respectively. The density of [3H]-InsP3 binding sites was 335.3 +/- 19.3 fmol mg-1 protein which was approximately 2.5 fold greater than that of [3H]-InsP4 sites (127.3 +/- 9.1 fmol mg-1 protein). 3. The two high affinity inositol phosphate binding sites exhibited markedly different pH optima for binding of each radioligand. At pH 9.0, specific [3H]-InsP3 binding was maximal, whereas [3H]-InsP4 binding was only 10% that of [3H]-InsP3. Conversely, at pH 5.0, [3H]-InsP4 binding was maximal, while [3H]-InsP3 binding was reduced to 15% of its binding at pH 9.0. 4. InsP3 was about 20 fold less potent (KI = 50.7 +/- 8.3 nM) than InsP4 in competing for [3H]-InsP4 binding sites and could compete for only 60% of [3H]-InsP4 specific binding. InsP4 was also capable of high affinity competition with [3H]-InsP3 binding (KI = 103.5 +/- 1.5 nM), and could compete for 100% of [3H]-InsP3 specific binding. 5. [3H]-InsP3 binding in subcellular fractions separated by discontinuous sucrose density gradients followed NADPH-cytochrome c reductase activity, suggesting an intracellular localization for the majority of InsP3 receptors in this tissue, whereas [3H] InsP4 binding appeared to be equally distributed between plasma membrane and intracellular membrane populations.6. These results suggest the existence of distinct and specific InsP3 and InsP4 binding sites which may represent the physiological receptors for these second messengers; differences in the subcellular distribution of these receptors may contribute to differences in their putative physiological roles. PMID- 8401944 TI - Characterization of CCK receptors in a novel smooth muscle preparation from the guinea-pig stomach by use of the selective antagonists CI-988, L-365,260 and devazepide. AB - 1. The cholecystokinin receptors mediating motor responses in a novel smooth muscle preparation from the corpus region of the guinea-pig stomach have been characterized by use of five agonist peptides and the antagonists CI-988, L 365,260 and devazepide. 2. Mucosa-denuded strips of circular muscle were contracted in a concentration-dependent manner by the five cholecystokinin (CCK) related peptides CCK-8S, pentagastrin, gastrin-I, CCK-8US and CCK-4. 3. CI-988 was a powerful antagonist of the response to pentagastrin with an affinity (pKB = 9.49) similar to that obtained in CCKB receptor binding assays. With CCK-8S as the agonist, CI-988 was approximately 1000 fold less powerful as an antagonist. 4. Devazepide powerfully blocked responses to CCK-8S with an affinity (pKB = 9.54) that was in agreement with reported functional data obtained in pancreatic amylase secretion studies, a system exhibiting CCKA receptor activity. Devazepide displayed lower affinity against pentagastrin than against CCK-8S. 5. CI-988 blocked responses to pentagastrin in an insurmountable manner in the presence of 3 nM devazepide; a concentration previously shown to block the CCKA receptor. The nature of the antagonism observed with L-365,260 was unaltered by the presence of devazepide. 6. The guinea-pig stomach corpus smooth muscle preparation contains both subtypes of CCK receptor and will be useful as a pharmacological tool for investigating the functional effects of novel CCK ligands. PMID- 8401945 TI - Block of voltage-dependent sodium currents by the substance P receptor antagonist (+/-)-CP-96,345 in neurones cultured from rat cortex. AB - 1. Whole cell patch clamp recordings of voltage- and tetrodotoxin-sensitive Na+ currents were made from cultured rat neocortical neurones (E18). The effects of the non-peptide NK1 receptor antagonist, (+/-)-CP-96,345 on Na+ currents was examined, relative to the effect of the local anaesthetic lignocaine and tetrodotoxin. 2. Sodium currents were reversibly depressed by bath application of (+/-)-CP-96,345 with a half-maximally effective concentration of 18 +/- 2 microM at a stimulation frequency of 0.1 Hz. Likewise the concentrations required to half-maximally inhibit sodium currents by tetrodotoxin and lignocaine were 10 +/- 2 nM and 1.3 +/- 0.2 mM respectively. 3. The depression of sodium currents by (+/ )-CP-96,345 (10 microM) was use-dependent in that raising the stimulus frequency from 0.1 Hz to 10 Hz further decreased the magnitude of sodium currents from 60 +/- 5% to 37 +/- 5% of control values respectively. Similarly, the depression of sodium currents by lignocaine (500 microM) and tetrodotoxin (30 nM) was also accentuated by raising the stimulus frequency from 0.1 Hz to 10 Hz. 4. The effect of (+/-)-CP-96,345 was not associated with a change in either the activation or steady-state inactivation characteristics of these currents, suggesting that its mechanism of action was via open channel blockade. 5. These data demonstrate that in addition to antagonizing NK1 receptors, (+/-)-CP-96,345 also acts as a channel blocker on sodium channels at micromolar concentrations, an effect which should be taken into consideration when examining the antinociceptive or anti inflammatory action of this compound. PMID- 8401946 TI - Effects of rubidium on responses to potassium channel openers in rat isolated aorta. AB - 1. In a physiological salt solution (PSS) in which potassium (K) was replaced by rubidium (Rb), segments of rat aorta precontracted with 20 mM RbCl were fully relaxed by K-channel openers with an order of potency levcromakalim > cromakalim > aprikalim > RP 49356. These relaxations were inhibited by glibenclamide. 2. Segments of rat aorta bathed in normal PSS and precontracted with 20 mM KCl were also relaxed by these K-channel openers with an order of potency levcromakalim > cromakalim > aprikalim > RP 49356. These relaxations were glibenclamide sensitive. However, the absolute potencies of the K-channel openers were approximately four times greater in normal PSS than in RbPSS. 3. In RbPSS, minoxidil sulphate relaxed segments of aorta precontracted with 20 mM RbCl by approximately 20% whereas in normal PSS it fully relaxed those contracted with 20 mM KCl. 4. In RbPSS, levcromakalim-induced relaxation of aortic segments precontracted with 20 mM RbCl was initially well-maintained but then faded by approximately 60% of the initial relaxation to a new, stable level. Subsequent exposure to RP 49356 or to higher concentrations of levcromakalim was without further relaxant effect. Similar changes were observed when RP 49356 was the initial relaxant and tissues were exposed to either RP 49356 or levcromakalim. In normal PSS, levcromakalim- or RP 49356-induced relaxation of contractions produced by 20 mM KCl was well-maintained. 5. In RbPSS, minoxidil sulphate induced relaxation of aortic segments precontracted with 20 mM RbCl was well maintained. Subsequent exposure to either levcromakalim or to RP 49356 produced further tissue relaxation. 6. In RbPSS, levcromakalim produced no detectable increase in either 86Rb- or 42K-efflux from rat aortic strips. In normal PSS, a significant increase in the exchange of both isotopes was detected.7. Levcromakalim hyperpolarized segments of rat aorta bathed both in normal PSS and after depolarization by the addition of 20 mM KCI. Exposure to RbPSS depolarized the tissue and under these conditions, levcromakalim had no effect on membrane potential.8. In Rb- and normal PSS, levcromakalim produced a similar degree of inhibition of the refilling of then or adrenaline-sensitive Ca store.9. It is concluded that millimolar concentrations of Rb inhibit the plasmalemmal ATP sensitive K-channels (KATP) which are the target of the K-channel openers. The relaxant actions of the K-channel openers in both normal and Rb-PSS and the inhibition of these effects by glibenclamide may reflect a functional interaction between these agents at ATP-binding sites associated with both KATP and with intracellular structures including Ca stores. PMID- 8401947 TI - Selective inactivation of muscarinic M2 and M3 receptors in guinea-pig ileum and atria in vitro. AB - 1. The role of muscarinic M2 and M3 receptors in ileal smooth muscle has been evaluated by use of selective receptor alkylation. The alkylating agents, 4 diphenylacetoxy-N-(2-chloroethyl)-piperidine (4-DAMP mustard) was studied for effects against (+)-cis-dioxolane, at muscarinic M2 and M3 receptors in guinea pig atria or ileum, respectively. 4-DAMP mustard (10 nM, 40 min exposure) did not discriminate between these muscarinic receptors. In ileum, 4-DAMP mustard, at 100 nM, resulted in a large dextral shift (197 fold) and depression in maxima. In atria there was a smaller dextral shift (14 fold) but no depression in maxima. 2. The muscarinic antagonists, atropine (non-selective), methoctramine (M2 selective) and para-fluorohexahydro-siladiphenidol (pFHHSiD; M3 selective) were studied in protection studies against alkylation by phenoxybenzamine. Washout studies following equilibration of the tissues with atropine (30 nM), methoctramine (0.3 microM) or pFHHSiD (3 microM), showed the compounds to be reversible. No temporal changes in sensitivity to (+)-cis-dioxolane were observed. 3. Exposure, for 20 min, of atria and ileum to phenoxybenzamine (3 and 10 microM respectively) caused dextral shifts and depressions in the maxima of the concentration-response curve to (+)-cis-dioxolane. These effects were inhibited by prior equilibration with atropine (30 nM) and methoctramine (0.1 microM) in atria or atropine (30 nM) and pFHHSiD (3 microM) in ileum. Similar results in ileum were obtained when pilocarpine was used as the agonist. 4. These data were consistent with muscarinic M2 receptors mediating responses in atria and M3 receptors mediating responses in ileum. No evidence was provided for a direct role of muscarinic M2 receptors in ileal contraction.5. It is concluded that receptor protection by reversible antagonists for muscarinic M2 or M3 receptors provides a means to isolate pharmacologically a single subtype in a tissue possessing heterogeneous populations. This technique may prove useful in defining the role of the respective subtypes in smooth muscle contraction. PMID- 8401948 TI - Bronchodilatation of guinea-pig perfused bronchioles induced by the H3-receptor for histamine: role of epithelium. AB - 1. The influence of epithelium on the effects of H3-histamine receptor agonist (R)alpha-methylhistamine [(R)alpha-MeHist] on airways was investigated on the guinea-pig perfused bronchioles. 2. In preparations under resting tone, removal of the bronchiolar epithelium or treatment with the cyclo-oxygenase inhibitor indomethacin (10(-5) M) increased the constriction induced by histamine and acetylcholine in a concentration-dependent manner without an alteration of the K(+)-induced contraction. 3. In this preparation (R)alpha-MeHist induced a concentration-dependent bronchodilatation which was antagonized in a competitive manner by thioperamide (an H3-antagonist) with a pA2 value of 8.6. 4. This bronchodilatation was reversed to a low concentration-dependent constriction after either removal of the epithelium or treatment with indomethacin (10(-5) M) but was unaffected by both 10(-5) M tranylcypromine (an inhibitor of PGI2 synthesis) and 5 x 10(-5) M NG-nitro-L-arginine methyl ester (an inhibitor of NO synthesis). 5. It is suggested that, in guinea-pig perfused bronchioles (R)alpha MeHist induces an epithelium-dependent relaxation via the release of metabolite(s) of arachidonic acid. PMID- 8401949 TI - Evidence that a form of ATP uncomplexed with divalent cations is the ligand of P2y and nucleotide/P2u receptors on aortic endothelial cells. AB - 1. The response of bovine aortic endothelial cells to adenosine 5'-triphosphate (ATP) is mediated by both P2y and nucleotide/P2u receptors. In order to determine which form of the nucleotide is the true ligand of these receptors, we have investigated the effects of divalent cations on ATP-, uridine 5'-triphosphate (UTP)- and 2 methylthioadenosine 5'-triphosphate (2MeSATP)-induced inositol phosphate accumulation in these cells. 2. Omisson of Mg2+ from a calcium-free incubation buffer caused a shift to the left of the ATP concentration-action curve. 3. In the presence of EDTA (1 mM), the basal level of inositol trisphosphate (InsP3) was markedly increased and the absolute maximal response to ATP was decreased; however, the response to low concentrations of ATP was enhanced. 4. When the results were plotted in terms of calculated ATP4- concentrations, the concentration-response curves obtained in the presence of 1.25 mM Mg2+ lay closer to the respective curves obtained when Mg2+ was omitted from the medium or when Mg2+ was omitted and EDTA (1 mM) was added. The curves became almost superimposable when the baseline value was subtracted. 5. A similar shift to the left of the concentrations-action curves was also observed with both UTP and 2MeSATP. 6. Our data provide evidence that a form of ATP uncomplexed with divalent cation is the preferential agonist of both the nucleotide/P2u and the P2y receptors expressed on bovine aortic endothelial cells. PMID- 8401950 TI - The effects of evening primrose oil on nerve function and capillarization in streptozotocin-diabetic rats: modulation by the cyclo-oxygenase inhibitor flurbiprofen. AB - 1. The aims of this study were first, to examine whether deficits in nerve conduction in streptozotocin-diabetic rats could be reversed by a 10% dietary supplement of evening primrose oil. Second, to determine the time-course of reversal, and third, to assess whether the effects could be blocked by the cyclo oxygenase inhibitor flurbiprofen (5 mg kg-1 day-1). 2. One-month diabetes produced 20% and 15% deficits in sciatic motor and saphenous sensory conduction velocity respectively, which were maintained over 2 months diabetes. 3. The effect of 1-month evening primrose oil treatment on abnormalities caused by an initial month of untreated diabetes was examined. Motor and sensory nerve conduction velocity were restored to the non-diabetic level. 4. Resistance to hypoxic conduction failure was investigated for sciatic nerve trunk in vitro. The 80% conduction failure times were 29% and 55% prolonged by 1- and 2-month diabetes respectively. Evening primrose oil did not reverse the increased hypoxic resistance following 1-month untreated diabetes. 5. Sciatic nerve endoneurial capillary density was not significantly affected by diabetes, but was 16% increased in diabetic rats with reversal by evening primrose oil treatment for 1 month compared to 2-month untreated diabetes. 6. Serial motor conduction velocity measurement after 3-month untreated diabetes revealed complete normalization by evening primrose oil within 4 days. Cessation of treatment resulted in a rapid decline in conduction velocity over 24 h. 7. In a preventive study of 2-month duration, 6 groups of rats were used. These comprised non-diabetic controls, diabetic rats, and evening primrose oil-treated diabetic rats, both with and without flurbiprofen treatment. Flurbiprofen had no significant effect in non diabetic rats, but produced an 11% worsening of motor conduction velocity and a 21% reduction of sciatic capillary density in diabetic rats. Evening primrose oil prevented the decreases in conduction velocity and increased hypoxic resistance with diabetes, and caused a 23% increase in capillary density. Flurbiprofen completely blocked the effect of evening primrose oil on conduction velocity, resistance to hypoxia, and capillarization.8. Six main conclusions were reached. First, evening primrose oil rapidly reverses conduction deficits in diabetic rats. Second, the effects of treatment may be very short-lived, suggesting a primary metabolic action. Third, evening primrose oil cannot reverse established changes in hypoxic resistance over 1-month treatment. Fourth, long-term treatment causes angiogenesis, suggesting a vascular action. Fifth,products of cyclo oxygenase-mediated metabolism are necessary for maintaining vasa nervorum integrity in diabetic rats. Sixth, evening primrose oil probably acts by providing substrate for vasodilator prostanoid synthesis by vasa nervorum. PMID- 8401951 TI - Vasoreactivity and prostacyclin release in streptozotocin-diabetic rats: effects of insulin or aldose reductase inhibition. AB - 1. Alterations in vasoreactivity and endothelial cell function could underlie some of the vascular abnormalities in diabetes. To examine aspects of these phenomena we studied the effects of 4-6 weeks streptozotocin-induced diabetes in the rat on basal and angiotensin II (AII)-stimulated prostacyclin release from isolated lung, perfused at constant flow. In addition, pressure was monitored throughout the lung perfusion as an index of vasomotor tone. 2. The experiment also included lungs from groups of diabetic rats treated with either insulin or an aldose reductase inhibitor (imirestat), to determine whether these treatments influenced the development of any defects seen in untreated diabetes. 3. Despite some indication of a trend towards reduced prostacyclin release in lungs from diabetic rats, neither the basal nor AII-stimulated release was significantly different from that seen in tissues from control animals. There were no significant differences between groups in the average basal perfusion pressure and in either the absolute pressure response to AII or the time of this peak. 4. The area under the perfusion pressure curve during AII infusion was greater in lungs from diabetic animals than in controls indicating a prolonged vasoconstrictor response. This increased pressor response may indicate increased sensitivity of diabetic tissue to AII or a reduced production of vasodilators in response to the vasoconstriction. 5. Whichever mechanism was responsible, this alteration was prevented by insulin treatment but not by aldose reductase inhibition, implicating mechanisms probably unrelated to exaggerated polyol pathway flux. PMID- 8401952 TI - Bacterial endotoxin rapidly stimulates prolonged endothelium-dependent vasodilatation in the rat isolated perfused heart. AB - 1. The effects of bacterial lipopolysaccharide (Escherichia coli 0111-B4; LPS) on coronary vascular tone were examined in the isolated perfused heart of the rat. The role of nitric oxide and/or prostaglandin products of the cyclo-oxygenase pathway in mediating the actions of LPS were also investigated. 2. Coronary vascular tone was raised and maintained by a continuous perfusion of the thromboxane-mimetic U46619 (5 nM). LPS perfusion (0.1-100 micrograms ml-1) caused a concentration-dependent fall in coronary tone without any significant change in the force of cardiac contractility. 3. At 5 micrograms ml-1, LPS reduced perfusion pressure by 38 +/- 9 mmHg. This effect was rapid in onset, maximal within the first 5 min and sustained for 90 +/- 10 min (n = 6). 4. The vasodilatation induced by LPS was dependent on the presence of an intact endothelium and abolished following endothelial damage caused by air embolism. 5. NG-nitro-L-arginine methylester (L-NAME; 50 microM) or NG-nitro-L-arginine (L NOARG; 50 microM) blocked the vasodilatation induced by LPS (5 micrograms ml-1). The inhibition caused by these arginine analogues was partially reversed by 1 mM L- but not D-arginine. 6. The vasodilator action of LPS was also completely blocked by the glucocorticoid, dexamethasone (10 microM) but unaffected by indomethacin (10 microM). 7. These results suggest that LPS evokes rapid release of nitric oxide (NO) in the microvasculature of the rat isolated heart presumably via activation of the constitutive L-arginine-NO pathway in the endothelium. Furthermore, the lack of effect of indomethacin suggests that prostaglandins released via the cyclo-oxygenase pathway are not involved in mediating this action of LPS. PMID- 8401953 TI - Acetylcholine-induced endothelium-independent relaxations in monkey isolated superior and inferior caval veins. AB - 1. We examined the effects of acetylcholine (ACh), isoprenaline (Isop) and Ca ionophore, A23187 on monkey isolated superior (SCV) and inferior caval veins (ICV) with and without intact endothelium, which had been partially contracted by 2 x 10(-6)-5 x 10(-6) M prostaglandin F2 alpha (PGF2 alpha). 2. Low concentrations of ACh (10(-10)-10(-9) M) produced a dose-dependent relaxation in the precontracted venous segments with endothelium. ACh at concentrations more than 10(-7) M elicited a transient contraction followed by a relaxation in these segments. 3. An addition of 5 x 10(-7) M A 23187 induced about 60% of maximum relaxation produced by 10(-5) M sodium nitroprusside (SNP) in each venous segment with endothelium. 4. Isop (10(-10)-10(-5) M) caused a dose-related relaxation in the precontracted caval veins with intact endothelium. 5. Removal of endothelium caused no significant effect on the ACh-induced dual responses but a significant inhibition of the A23187-induced relaxation. 6. Pretreatment with atropine antagonized competitively the ACh-induced relaxations in the endothelium-intact and endothelium-denuded caval veins. The Schild plot analysis showed that the pA2 values of the segments with and without endothelium were 9.72 +/- 0.14 (n = 5) and 10.01 +/- 0.23 (n = 6) in the ICV; and 9.95 +/- 0.20 (n = 5) and 9.70 +/- 0.10 (n = 5) in the SCV, respectively. 7. Pretreatment with 5 x 10-5M aspirin, 3 x 10-5M N0-nitro-L-arginine methylester, 1 mM tetraethylammonium,or 3 x 10-6 M glibenclamide caused no significant effect on the basal tone, ACh induced transient contraction, and ACh;.induced relaxation in the precontracted venous segments with and without endothelium.8. Pretreatment with 10-5 M methylene blue produced a significant reduction of the ACh- and SNP induced relaxations in the precontracted venous segments with and without endothelium. The pretreatment with the same concentration of methylene blue, however, caused no significant effect on the Isop-induced relaxation in venous segments with endothelium.9. The results suggest that ACh acts directly on the venous smooth muscle cells via a high affinity muscarinic receptor subtype to accumulate cellular cyclic GMP producing endothelium-independent relaxation in the monkey caval veins. PMID- 8401954 TI - Heterogeneity of muscarinic receptors in lamb isolated coronary resistance arteries. AB - 1. In vitro experiments in a microvascular myograph were designed to characterize postjunctional muscarinic receptors producing contraction both in the presence and absence of the endothelium in coronary resistance arteries (normalized diameter of 150-450 microns), isolated from the left ventricle of hearts from 3-6 month old lambs. Preferential muscarinic receptor antagonists were used to determine the receptor subtype: pirenzepine (M1 receptor), AFDX 116 (M2 receptor), 4-DAMP and pFHHSiD (M3 receptor). 2. The rank order of potency for muscarinic agonist-induced increases in tension in endothelium-intact preparations was oxotremorine-M = methacholine = acetylcholine (ACh) > carbachol. Removal of the endothelium increased the potency of ACh, but this procedure did not change either the sensitivity or maximal response to carbachol. 3. The contractile response to ACh was reproducible. Incubation with 3 x 10(-7)-3 x 10( 6) M pirenzepine induced non-parallel rightward shifts and depressed the maximum of the concentration-response curve to ACh in endothelium-intact arteries. The slope by Schild analysis was 2.9 +/- 0.8 (P < 0.05, n = 7). Atropine, AFDX 116, 4 DAMP and pFHHSiD produced parallel rightward shifts of the curves to ACh and the slopes of the Schild plots were not significantly different from unity. The pKB values for the antagonists from plots constrained to unity in endothelium-intact segments were: atropine (9.4), 4-DAMP (9.0), pFHHSiD (7.9) and AFDX 116 (6.2). 4. In endothelium-denuded arteries, pirenzepine, AFDX 116 and pFHHSiD caused concentration-dependent, parallel rightward displacements of the concentration response curves to ACh and the slopes of the Schild plots were not significantly different from unity. The plots constrained to a slope of unity gave the following pKB values: pFHHSiD (8.7), pirenzepine (7.5) and AFDX 116 (6.2). 5. In the presence of the endothelium, low concentrations of pirenzepine (10(-9)-10(-7) M) produced leftward shifts of the ACh concentration-response curves. This potentiating effect of pirenzepine was reversed by endothelial cell removal. In preparations precontracted with the thromboxane-mimetic, U46619, the putative M1 selective agonist, McN-A-343, induced a biphasic relaxation with log IC50 of 8.53 +/- 0.14 and 5.02 +/- 0.08 for the first and second phase of the relaxation, respectively, and maximal relaxations of 22.8 +/- 4.3% and 41.1 +/- 5.4% (n = 16). McN-A-343 relaxed the vessels in the presence of 10(-7) M pFHHSiD and 3 x 10(-7) M AFDX 116, but not after incubation with 10(-9) M pirenzepine. 6. It is concluded from the pKB values for the antagonists used, that contraction induced by ACh in lamb coronary resistance arteries, in either the presence or the absence of the endothelium, is mediated via the M3 subtype of muscarinic receptors, while muscarinic receptors of another subtype at the endothelium seem to modulate the contractile response to ACh. PMID- 8401955 TI - Psychiatric aspects of heart transplantation. AB - There are many practical psychiatric, social, and ethical problems which accompany heart transplantation. These include pre- and postoperative anxiety and depressive conditions, post-operative delirium, and social and family dysfunctional syndromes. This paper reviews the literature critically in the following five areas: pre-transplant evaluation, coping with surgery, postoperative sequelae, rehabilitation, and management. Although most recipients have a good outcome from the physical and psychiatric points of view, a substantial minority experience family conflicts and sexual dysfunction after surgery. An average of 45% of patients from all studies returned to full-time employment. The distinctive role and contribution of the psychiatrist on the transplant team is discussed and important areas for future research are outlined. PMID- 8401956 TI - What do depression rating scales measure? AB - A large number of rating scales have been devised to assess the clinical construct of 'depression'. These scales have been universally used in research with little consideration of their content, or how they relate to accepted definitions of depressive disorder. The scales are often arbitrarily selected and used for the study on the assumption that all measure the same construct. The item analysis of a number of the most widely used depression scales reveals a variation in the areas of psychopathology they cover; some scales place greater emphasis upon the assessment of anxiety than upon depressed mood. Since disturbance in neurobiological systems is manifest by specific aspects of affective and behavioural malfunction, and since psychodynamic factors lead to particular cognitive sets, the advancement of research will depend upon the construction and validation of more refined measures than are provided by the present approach. PMID- 8401957 TI - Genotypes, phenotypes, and the brain. A search for connections in schizophrenia. PMID- 8401958 TI - Recognising psychiatric symptoms. Relevance to the diagnostic process. AB - Current overemphasis on nosological diagnosis has led to a neglect of the process of symptom recognition. There is evidence, however, that the perception of the symptom alone does not guarantee symptom ascertainment since a decision-making component is also involved. To achieve the latter, additional information must be provided by the contextual cues implicit in the ongoing diagnostic hypothesis. Current diagnostic systems, however, still assume a two-stage model according to which symptom and disease recognition are independent cognitive events. This paper suggests that this model is inadequate and that descriptive psychopathology is not transparent. It then describes a neural network simulation to make various aspects of the problem explicit. This takes into account the multidimensional and probabilistic aspects of symptom recognition and is, from this point of view, superior to traditional algorithmic models. It also has the capacity to represent the different cognitive styles involved in symptom recognition. PMID- 8401959 TI - A randomised controlled trial of psychotherapy in patients with refractory irritable bowel syndrome. AB - Patients with chronic, refractory irritable bowel syndrome (n = 102) were entered into a randomised controlled trial of psychotherapy versus supportive listening. Independent physical and psychological assessments were carried out at the beginning and end of the 12-week trial. For women, psychotherapy was found to be superior to supportive listening, in terms of an improvement in both physical and psychological symptoms. There was a similar trend for men, but this did not reach significance. Following completion of the trial, patients in the control group were offered psychotherapy; 33 accepted and following treatment experienced a marked improvement in their symptoms; ten declined. At follow-up one year later, those patients who had received psychotherapy remained well, patients who had dropped out of the trial were unwell with severe symptoms, and most of the controls who declined psychotherapy had relapsed. This study shows that psychotherapy is feasible and effective in the majority of irritable bowel syndrome patients with chronic symptoms unresponsive to medical treatment. PMID- 8401960 TI - Seasonal variations of current symptoms in a healthy population. AB - Among a large workplace population interviewed over a year, current symptoms were assessed using the Hopkins Symptom Checklist (HSCL). Variation in symptoms by date of assessment was observed among the 314 women but not the 1556 men. Among women, symptoms were greatest during late autumn and winter, and significant inverse correlations were found between available daylight at the time of assessment and standard symptom dimensions of anxiety and somatisation, as well as an expanded mood scale more inclusive of depressive symptoms within the check list. The amplitude of the seasonal effect was such that the prevalence of female 'cases', as defined by HSCL criteria, was twice as high during winter than during the rest of the year. The data are consistent with a role for light-dark exposure in eliciting or synchronizing annual mood rhythms. The sex-by-season interactions may contribute to the sex differences in overall prevalence of depression. PMID- 8401961 TI - Seasonal affective disorder: response to light as measured by electroencephalogram, melatonin suppression, and cerebral blood flow. AB - This study was designed to test the hypothesis that patients with SAD have significantly different physiological responses to light than healthy age- and sex-matched controls. We studied retinal contrast sensitivity, visual evoked EEG responses, and melatonin suppression by, and cerebral blood flow response to, full-spectrum artificial daylight. There was no significant difference between 10 patients and 11 controls in retinal contrast sensitivity, or amplitude or latency of N2, P2, P100 or P300 on EEG. We compared melatonin suppression in 12 SAD patients and 12 controls. During exposure to 500 lux and 1500 lux artificial daylight both the SAD patients and controls had a significant melatonin percentage suppression; however, the percentage suppression did not differ significantly between the SAD patients and the controls. In addition, we carried out a small pilot study into the effect of light on cerebral blood flow in four SAD patients and four controls. Before exposure to 1500 lux artificial daylight there was no significant difference between patients and controls in global, regional or cerebral hemispheric blood flow. After light exposure the SAD patients and controls had a significantly different percentage change in cerebral blood flow. We suggest that patients with SAD do not have significantly physiological responses to light than controls, except perhaps in cerebral blood flow. Furthermore, mechanisms other than supersensitivity of melatonin suppression must explain both the pathophysiology of the disorder and its response to treatment with light. PMID- 8401962 TI - Melatonin rhythms in seasonal affective disorder. AB - We examined 24-hour melatonin rhythms from 20 patients with seasonal affective disorder (SAD) and 20 healthy volunteers. Patients and controls were individually matched for age, sex, and month of study. Plasma samples were taken at hourly intervals, and were assayed for melatonin by radio-immunoassay. The 24-hourly melatonin estimations for each individual were fitted to a cosine curve, and the significance of the curve fits was calculated. Two analyses were performed. In analysis 1 the following were calculated: (a) cosine fit, (b) significance of fits, (c) mean amplitude and acrophase (peak) and (d) mean melatonin levels. The curve fits were highly significant for all but three subjects (two patients, one control), but there were no significant differences in any measure between the two groups. In analysis 2 the comparisons were repeated and restricted to the 18 patients and 19 controls in whom there was a significantly significant melatonin rhythm. Again there were no significant differences between groups. These results suggest that the circadian rhythm of melatonin is not abnormal in SAD, and that the therapeutic effect of light in SAD is not mediated by phase shifts in melatonin secretion. PMID- 8401963 TI - Cognitive impairments in patients with seasonal affective disorder. AB - Eleven patients with seasonal affective disorder (SAD) and ten controls matched for age, IQ, and education were tested on a number of computerised tests designed to assess attention, memory, and learning. When depressed, patients showed no deficit in attention but were impaired on spatial memory and learning. They were also significantly slower to respond than controls, with a pattern that suggested slowed information processing centrally, rather than simple sensory or motor slowing. On recovery from depression, improvement was seen in most tests, although impairment remained in latency to respond on a test of spatial memory. This continuing impairment correlated with residual depressive symptoms but not with ventricular brain ratio (VBR). PMID- 8401964 TI - Positive, negative, and disorganisation factors from the Schedule for Affective Disorders and Schizophrenia and the Present State Examination. A three-factor solution. AB - The use of items from the Schedule for Affective Disorders and Schizophrenia and from the Present State Examination scales for assessing positive and negative symptoms in schizophrenia was examined using factor analysis. The factorial structure of the items which putatively assess positive and negative symptoms was examined. A three-factor solution was obtained with factors identified as: a negative symptom factor (factor 1); a positive symptom factor (factor 2); a 'disorganisation' factor (factor 3), consisting primarily of items related to disordered thinking. A solution which was highly similar in important loadings was obtained with an independent sample of patients. High correlations of the rotated factors with the external criteria supported the interpretations of the factors. The results indicate that symptoms generally classified as negative or positive are factorially independent. Furthermore, a disorganisation factor, consisting of items previously included in positive and negative symptoms factors, is necessary for a full representation of the factor structure. PMID- 8401965 TI - A comparative study of 470 cases of early-onset and late-onset schizophrenia. AB - The presence or absence of 22 schizophrenic symptoms was recorded with the age at onset of illness in 470 patients with non-affective, non-organic psychoses. Positive and negative formal thought disorder, affective symptoms, inappropriate affect, delusions of grandiosity or passivity, primary delusions other than delusional perception, and thought insertion and withdrawal were all more common in early-onset cases (age at onset 44 years or less; n = 336). Persecutory delusions with and without hallucinations, organised delusions, and third-person, running commentary and accusatory or abusive auditory hallucinations were all more common in late-onset cases (age at onset 45 years or more; n = 134). There was no difference between cases of early and late onset in the prevalence of delusions of reference, bizarre delusions, delusional perception, or lack of insight. We conclude that although there are clinical similarities between cases of schizophrenia with early and late onset, there are sufficient differences between them to suggest that they are not phenotypically homogeneous. PMID- 8401966 TI - Effects of a history of heavy alcohol consumption on Alzheimer's disease. AB - Neuropsychological and psychiatric evaluations were made of 39 subjects with possible Alzheimer's disease and a history of excessive alcohol consumption (AD + ETOH), who had been abstinent or had drunk minimally for at least three months before evaluation, and 225 patients with probable Alzheimer's disease (PAD) of comparable age, years of education, and baseline global impairment. At baseline, there were no significant differences between the groups in terms of age of onset of dementia, neuropsychological test scores, or current behavioural or psychiatric symptoms. One year later, no differences in rates of decline between 20 abstinent AD + ETOH patients and 88 PAD subjects could be shown. Thus, past heavy alcohol consumption does not appear to modify the presentation of dementia of the Alzheimer's type, nor does it modify progression over a one-year interval. PMID- 8401967 TI - Neuropathological correlates of behavioural disturbance in confirmed Alzheimer's disease. AB - Clinico-pathological correlations were examined in 54 patients with neuropathologically verified Alzheimer's disease (AD) who were part of a prospective study. Behavioural disturbance was documented using an expanded version of the Stockton Geriatric Rating Scale (SGRS). The subscores for physical disability (P), apathy (A) and communication failure (C) (summation score PAC) were closely correlated and were high in most patients during the late stages of illness. High PAC scores correlated with an earlier onset and longer duration of illness, lower brain weight, more severe tangle pathology in the parahippocampal gyrus and the frontal and parietal neocortex, and lower neuron counts in the hippocampus and basal nucleus of Meynert. Features of the Kluver-Bucy syndrome (range behaviour and hypermetamorphosis) were significantly associated with lower counts of large neurons in the parahippocampal gyrus and parietal neocortex, but not with more severe plaque or tangle formation or with neuronal loss in the subcortical nuclei. No correction was made for multiple comparisons. These findings may signify decreased cortical inhibition in patients with relatively well preserved subcortical structures who show features suggestive of the Kluver Bucy syndrome. High PAC scores on the SGRS could reflect more advanced and widespread cerebral pathology in the end stages of AD. PMID- 8401968 TI - A community study of depression in adolescent girls. I: Estimates of symptom and syndrome prevalence. AB - A community study was conducted to determine the prevalence of depressive symptoms and syndromes in 11-16-year-old girls. Girls from three secondary schools (1072 girls in total) were administered questionnaires and 375 were selected for direct interview. The estimated prevalence for current major depressive disorder (within the past month) for the population was 3.6%, and for the past year was 6.0%. Scores on the self-report questionnaire of mood disturbance increased with age, as did the prevalence of depressive disorder. A large proportion of interviewed girls reported depressive symptoms which did not meet the DSM-III-R criteria for major depressive disorder. The estimated prevalence of this 'partial syndrome' group was 8.9% for a current episode and 20.7% for the past year. PMID- 8401969 TI - A community study of depression in adolescent girls. II: The clinical features of identified disorder. AB - The clinical characteristics of two subsamples of 11-16-year-old girls were determined from direct interview: those who met DSM-III-R criteria for an episode of major depressive disorder within the past month (n = 28); and those who did not currently meet these criteria but had done so at some time in the previous 12 months (n = 13). The symptom profiles of these cases were compared with a subsample of girls who reported depressive symptoms but did not meet DSM-III-R criteria and were designated as having a 'partial syndrome' (n = 93), and a sample of non-depressed controls (n = 129). Phobias and worry about peer acceptance were common in controls suggesting that these symptoms constitute normal adolescent concerns. The symptom profile of depressed cases altered across three age bands (11-12, 13-14, and 15-16 years), suggesting developmental influences on clinical presentation. Comorbidity for anxiety, behavioural, and obsessional disorders was found in 40% of the depressed cases. None of the cases of major depressive disorder was known to the clinical services. PMID- 8401970 TI - Life events and relapse in established bipolar affective disorder. AB - A New Zealand cohort of 58 patients with bipolar affective disorder was studied prospectively with three-monthly interviews in order to determine the relationship between life events and their relapses. Careful attention was paid to dating life events and the earliest signs of relapse and to assessing the independence of life events from the illness. No statistically significant association was found between life events and the likelihood of relapses, either mania or depression, for the 71% of patients who experienced at least one relapse during the two-year study. This finding is at variance with a companion study, with identical methodology, which found a small increase of life events before relapse. These data add further weight to the previous reports that life events are significant precipitants of bipolar illness only for earlier episodes in the course of this chronic disorder. PMID- 8401971 TI - Self-mutilation in four historical cases of bulimia. AB - Current theories suggest that there is a relatively frequent association of self mutilative behaviour with eating disorders, particularly with the modern binge purge syndrome, bulimia nervosa. In order to consider this association on a historical dimension, 25 bulimic cases, reported from the late 17th to the late 19th century, were investigated. These were found to include four examples of self-mutilative behaviour, in three males and one female: these cases are described and discussed. The historical evidence lends some support for the suggested connection between eating pathology and self-mutilation. PMID- 8401972 TI - A hormonal component to postnatal depression. PMID- 8401973 TI - The essential psychotherapies. PMID- 8401974 TI - The essential psychotherapies. PMID- 8401975 TI - The essential psychotherapies. PMID- 8401976 TI - Lithium neurotoxicity at normal therapeutic levels. PMID- 8401977 TI - Court diversion. PMID- 8401978 TI - Integrated psychological therapy for schizophrenia. PMID- 8401979 TI - Offspring of parents with drinking problems. PMID- 8401980 TI - Mania and Down's syndrome. PMID- 8401981 TI - Low serum cholesterol and serotonin receptor subtypes. PMID- 8401982 TI - Concepts of illness and disease. PMID- 8401983 TI - Neurological complications of anorexia nervosa. PMID- 8401984 TI - Abstraction of covariation in category learning: a critical note on K. Richardson's studies. AB - Several studies directed by K. Richardson (Richardson, 1986, 1987; Richardson & Carthy, 1989, 1990) claimed that people are able to abstract imperfect covariations among feature variables defining artificial categories, and to use that information in various subsequent tasks. We challenge this claim on a twofold basis. First, a large part of the performance variance Richardson attributed to covariation knowledge may be accounted for by a very simple exemplar model, which assumes no abstractive processes. Second, the residual influence of relational information on performance may be attributed to the knowledge subjects have acquired in real-world situations before their training with the study exemplars. PMID- 8401985 TI - Adolescent identity and school type. AB - The study examined the Eriksonian notion of exploration of identity alternatives and commitment to goals and beliefs during adolescence, focusing on whether this was associated with school type. A sample of 127 girls from one private and one state school, drawn from comparable social-class backgrounds, were interviewed in depth. Each subject was classified according to Marcia's (1966) four categories of identity achievement, moratorium, foreclosure and diffusion in the domains of politics and occupation. Significant differences between the private- and state educated samples are reported for both identity exploration and commitment in the two domains. Possible explanations for these differences are given, and recommendations for future work are made. PMID- 8401986 TI - Alcohol, reaction time and memory: a meta-analysis. AB - Moderate doses of alcohol impair performance on a variety of information processing tasks. Two separate meta-analyses were conducted on the results of (1) reaction time studies, and (2) recognition memory studies, representing 25 and 16 different task conditions, respectively. In both cases, performance with alcohol (either 0.8 or 1.0 ml/kg body weight) was plotted as a function of performance with no alcohol. For reaction time, a linear fit accounted for 99.7 per cent of the variance. The same function applied not only to the mean but to the distribution of reaction times from the 5th to the 95th percentiles. For recognition memory, a linear fit accounted for 96.2 per cent of the variance in accuracy (expressed as the logarithm of proportion correct). Thus alcohol appears to have a general linear effect on information processing, rather than specific effects on a subset of stages. It is concluded that the results are consistent with a reduced processing resources hypothesis for the impairment with alcohol. PMID- 8401987 TI - Human instrumental learning: a critical review of data and theory. AB - This article reviews experimental data from human instrumental learning tasks in which people acquire knowledge about the consequences of their actions. The main part of the paper examines the stimulus conditions which appear to control the acquisition of instrumental knowledge. These conditions include contiguity between the action and outcome, the degree of contingency between them, and also the extent to which the action is a good relative predictor of the outcome. Several accounts are examined of the mechanism by which instrumental knowledge might be acquired, including: (i) a variety of rule-based models, in which learning consists of the acquisition of knowledge about statistical relationships between contingent events; (ii) a relative contiguity model, in which learning involves the acquisition of knowledge about temporal relationships; and (iii) an associative model, in which learning involves the formation of mental associations which are updated by a learning rule. The review indicates that at present, the data seem most consistent with the associative learning model. However, there remain empirical phenomena which have resisted theoretical analysis. A variety of questions which future research might profitably explore are considered. PMID- 8401988 TI - The cognitive profile in infantile autism--a study of 70 children and adolescents using the Griffiths Mental Development Scale. AB - Seventy children and adolescents with infantile autism were tested with the Griffiths Mental Development Scale II. A characteristic profile emerged with peaks in motor and visuo-spatial domains and troughs in verbal and practical reasoning areas. These results support conclusions from previous research using other cognitive tests. PMID- 8401989 TI - Selective influence of test anxiety on reading processes. AB - The aim of this research is to explore whether comprehension is impaired by test anxiety, whether the anxious person spends compensatory reading time, and which cognitive processes are affected. High- and low-anxiety students read texts word by word with the moving-window technique under test conditions. Multiple regression analyses on word-reading times were computed with a number of psycholinguistic variables--assumed to map onto specific processes--serving as predictors. Results indicated that anxiety did not impair comprehension, but increased word-reading times, which were affected interactively by anxiety and specific psycholinguistic variables (end of clause, serial position within the text, narrativity, and summary). These data reveal that anxious readers need to employ a greater amount of processing resources than their non-anxious counterparts to obtain a similar comprehension level. Furthermore, the interactive effects suggest that anxiety is selectively detrimental to the efficiency of text-level processes, such as those involved in integrating information across sentences. In contrast, anxiety does not impair low-level processes, such as encoding and lexical access. PMID- 8401990 TI - Exploring the limits of optimism: the case of smokers' decision making. AB - Smokers consider themselves less likely than others to contract smoking associated diseases. This result has been interpreted as a weak form of irrationality. Alternatively, it can be interpreted as indicating that smokers show an optimism bias. The optimism explanation was examined by having smokers and non-smokers rate the likelihood of future negative events falling into three categories: (1) smoking associated, (2) health related and (3) health unrelated. Subjects rated their likelihood of the event happening to (1) themselves, (2) the average smoker and (3) the average non-smoker. Smokers and non-smokers rated their own likelihood of negative events as less than for the average smoker and the average non-smoker. But, while smokers rated their own risk as higher for both smoking-associated and other health-related problems, they rated their own risk as lower than the average smoker, a standard optimism bias. PMID- 8401991 TI - The grief process and job loss: a cross-sectional study. AB - This study assessed the applicability of the grief process to job loss. A pilot study of 10 unemployed men was used to establish a structured interview, and its content analysis, based on a description of the grief process derived from studies of bereavement. Other measures assessed the degree of attachment to the former job. The main study involved 60 men who had lost their jobs during the previous eight years. Individual grief items were found in some of these people, varying in frequency from 10 to 80 per cent. Principal components analysis revealed a general grief component, representing most of the specific items. Twenty-seven per cent of the sample fulfilled a criterion for a clear grief-like response. An overall grief score based on the interview answers was significantly correlated with three different measures of job attachment, and also with questionnaire measures of depression and anxiety. These measures were unrelated to the length of time since job loss, apparently providing no support for stage theories or for more general assumptions of adaptation. However, the cross sectional nature of the sample complicates this conclusion. PMID- 8401992 TI - Behavioural problems associated with the chronic fatigue syndrome. AB - Disturbances of memory, concentration and motor function are often reported by patients with the chronic fatigue syndrome (CFS). The present study objectively evaluated these behavioural problems using a computerized test battery measuring memory, attention and motor skills. Fifty-seven CFS patients were compared with 19 matched controls and all subjects completed the performance test battery and filled in questionnaires measuring psychopathology and mood. The patients reported significantly higher levels of depression, anxiety, physical symptoms and cognitive failures than the controls. Similarly, they reported more negative affect at the time of testing. The patients were slower on psychomotor tasks, showed increased visual sensitivity and impaired attention. Digit span and free recall were not impaired but retrieval from semantic memory and logical reasoning were slower. None of the performance differences between patients and controls could be attributed to differences in psychopathology. These results agree with recent findings from other laboratories, and it is now time to consider the nature of the neurological dysfunction underlying these effects. PMID- 8401993 TI - What is the future for the genetics of rheumatoid arthritis? PMID- 8401994 TI - The ABC of the diabetic hand--advanced glycosylation end products, browning and collagen. PMID- 8401995 TI - A quantitative ELISA for measurement of rheumatoid factor associated cross reactive idiotypes in serum from patients with rheumatic diseases. AB - A sensitive ELISA for the quantitation of RF associated cross-reactive idiotypes (CRI) has been developed. CRI bearing IgM was captured by immobilized F(ab')2 fragments of monoclonal anti-CRI antibodies and revealed with isotype specific conjugates. Factors affecting sensitivity, specificity and reproducibility of the assay were assessed. The sensitivity limit was less than 10 ng/ml and the linear assay range for the CRI standard curve was from 80 to 500 ng/ml. Using this ELISA system we have quantitated serum levels of the CRI in different groups of patients with RA, early synovitis and normal individuals. Our results indicate a significant increase in the levels of the CRI in the patient groups compared to the normal subjects tested, and suggest involvement of a diverse repertoire of Ig variable region germline genes and/or somatic mutation in production of polyclonal RF in RA. PMID- 8401996 TI - HLA-DQ, DR and complement C4 variants in systemic lupus erythematosus. AB - We have defined HLA-DQA, DQB, DR and complement C4 variants in 92 subjects with SLE and 73 controls. Subjects with SLE showed an increased frequency of HLA DQA*0501 (P < 0.01 corrected, odds ratio (OR) = 4.97; 95% C.I. = 2.52-9.81), DR3 (P < 0.001, OR = 3.18; 95% C.I. = 1.67-6.04) and C4A*Q0 (P < 0.05, OR = 1.91; 95% C.I. = 0.999-3.65) vs controls. These increases were particularly marked in those subjects positive for antibodies to both Ro and La. HLA-DQB*0501 (P < 0.01 corrected, OR = 0.03), DQA*0101 (P = 0.0012 uncorrected, OR = 0.23) and DR7 (P = 0.0018 uncorrected, OR = 0.28) were decreased in frequency in SLE. SLE patients with disease onset prior to age 30 yr were more likely to possess a DR3-bearing haplotype (P < 0.05 corrected) than those with onset after age 30 yr. No significant associations were found in patients with circulating antibodies to double-stranded DNA, Ro alone, U1 RNP, Sm or in those SLE patients with renal disease or vasculitis. The different associations found in different clinical and immunological subsets of SLE support the concept that SLE contains a variety of immunogenetic subgroups. Analysis of the associations between SLE and DR3, DQA*0501 and C4A*Q0 using the empirical logistic test suggests that the association of SLE with HLA-DQA*0501 is likely to be primary to the associations with both DR3 and C4A*Q0 (P < 0.001). Our results therefore raise the possibility that genes within the HLA-DQ region may have a direct effect upon susceptibility to SLE. PMID- 8401997 TI - The differential expression of heat shock proteins in rheumatic disease. AB - The concept of overexpression of endogenous heat shock proteins (hsps) is central to hypotheses in which hsps are implicated in the pathogenesis of autoimmune rheumatic disease. Hsps were quantitated in protein samples prepared from peripheral blood mononuclear cells (PBMC) of patients and controls by Western blotting and scanning densitometry. Evidence is presented for a specific pattern of overexpression of the 90 kDa hsp (hsp 90) and the highly inducible member of the 70 kDa hsp family (hsp 72) occurring overall in SLE relative to other diseases. Evidence is also presented for the differential expression of individual hsps in other autoimmune rheumatic diseases such as RA, primary SS and systemic sclerosis, and in other relevant conditions such as infectious diseases and multiple sclerosis. It is concluded that, while hsp 90 overexpression may have a specific role in, for example, SLE, overexpression of hsp 72 and underexpression of the constitutive member of the hsp 70 family (hsp 73) may be a more general reflection of ongoing disease states. PMID- 8401998 TI - Both inherited HLA-haplotypes are important in the predisposition to rheumatoid arthritis. AB - The distribution of the HLA-DR allele frequencies of 105 RA patients has been compared with the expected distribution under recessive and dominant modes of inheritance using control data from 2041 controls and the antigen genotype frequency among patients methodology. The observed distribution was compatible with a recessive mode of HLA-linked inheritance in RA, with a dominant mode rejected, whether HLA-DR4 was considered alone, or HLA-DR4 and HLA-DR1 were combined as if they were behaving as a single predisposing gene. Mean sibship concordance rates (MSCRs) were calculated for categories of proband HLA-DR genotypes. The highest MSCR was for HLA-DR4 homozygous probands, and the lowest for HLA-DR2 or 7/non-4 genotypes. These combined observations suggest that interactions between both inherited HLA-haplotypes are important in the predisposition to RA. PMID- 8401999 TI - Major histocompatibility complex variants and articular disease severity in rheumatoid arthritis. AB - In the light of associations previously described between DR4, DQw7, the C4B null allele and Felty's syndrome, and between Dw14, the C4A null allele and rheumatoid vasculitis, we have looked for associations between these and other DRB1, DQB, DQA and C4 encoded variants, and articular disease severity assessed radiologically in 119 subjects with RA but without major extra-articular features. The association of DR4 with more severe disease and DR1 and DR2 with mild disease was confirmed but no associations were found between the presence of DQw7, Dw14, C4A or C4B null alleles and articular disease severity. This suggests that the associations noted previously between these markers and extra-articular disease features of Felty's syndrome or rheumatoid vasculitis are unlikely to reflect an overall effect on disease severity but more likely to represent associations with these particular extra-articular disease subsets. PMID- 8402000 TI - Twin concordance rates for rheumatoid arthritis: results from a nationwide study. AB - We report the concordance rate for RA in a nationwide study of 91 monozygotic (MZ) and 112 dizygotic (DZ) pairs. Twin pairs were recruited from both a national media campaign and a 2-month prospective inquiry of all UK rheumatologists. Disease status was established following a structured clinical and serological appraisal, together with radiological assessment where necessary. Zygosity was confirmed using DNA fingerprinting. In all, 14 (15.4%) of the MZ and four (3.6%) of the DZ pairs were disease concordant (risk ratio: 4.3 95% CI 1.5 to 12.6). There was no difference in the concordance between the media and clinical derived twins. Further the excess MZ concordance persisted after adjusting for age, age at disease onset, sex and rheumatoid factor status. Analysing the data in relation to the timing of disease onset in the first affected twin showed that subsequent disease risk in the initially unaffected co-twins of the MZ affected probands increased with increasing duration of follow-up. We conclude that the overall MZ concordance at 15% is lower than the 30% figure normally quoted from a study some 30 years ago and sets a ceiling at the potential genetic contribution to disease susceptibility. PMID- 8402001 TI - Height, occupation and back pain in a national prospective study. AB - Back pain is an important public health problem but there is a paucity of knowledge about risk factors and causal mechanisms. Previous studies have shown that tall men are more at risk of back pain, although observations in women have been less consistent. This paper presents findings from a national longitudinal study of 3262 men and women aged 43 yr. Standing height and sitting height were related to 18-month reported prevalence of 'sciatica, lumbago or severe backache' in both men and women. The paper investigates explanations for these findings using previously collected data on childhood growth and detailed lifetime occupational histories. Neither greater susceptibility of tall men to heavy lifting, nor the timing of growth, were able to account for these relationships. To assess further the association between height and back pain, information is needed on the relationship between stature and characteristics of spinal structure. PMID- 8402002 TI - Magnetic resonance imaging manifestations of idiopathic haemochromatosis in the wrist. AB - The magnetic resonance imaging features of the wrist of a patient suffering from the arthropathy of haemochromatosis are presented. It is apparent that the deposition of iron within the bone marrow is focal in origin and may be associated with cyst formation. In addition, changes in serum ferritin levels with treatment suggest that the deposition is irreversible. Studies of two other patients with haemochromatosis without cyst formation in the wrists did not yield similar artefacts, in spite of having high ferritin levels and arthritis. PMID- 8402003 TI - Features of systemic sclerosis (scleroderma) in an identical twin pair. AB - The aetiology of systemic sclerosis (scleroderma) is unknown but it is thought to have both genetic and environmental components. The familial incidence of the disease is very low and we have been able to find only one report of scleroderma in identical twins which was in the Russian literature. We report here on a set of identical twins and their mother who all had features of systemic sclerosis. PMID- 8402004 TI - Idiopathic polymyositis complicated by arthritis and mesangial proliferative glomerulonephritis: case report and review of the literature. AB - A 56-yr-old male developed a systemic illness while receiving cyclical oral etidronate therapy for idiopathic osteoporosis. The illness, characterized by fever, proximal myopathy and inflammatory synovitis, was associated with interstitial lung disease and mesangial proliferative glomerulonephritis. Elevated plasma creatine phosphokinase level and inflammatory muscle biopsy findings confirmed a diagnosis of polymyositis (PM). Antibodies to Jo-1 were also detected. A review of the literature reveals that mesangial proliferation is the commonest glomerular lesion and suggests a possible association between arthritis and glomerulonephritis in PM. The prognosis of this renal lesion appears to be good, although only limited data is available. PMID- 8402005 TI - Malignant 'angioendotheliomatosis'--(intravascular lymphomatosis) an unusual cutaneous lymphoma in rheumatoid arthritis. AB - A case of cutaneous malignant 'angioendotheliomatosis' (intravascular lymphoma) is described in a patient with RA and Sjogren's syndrome. The association of RA with this rare form of lymphoma has not been reported in the literature. PMID- 8402006 TI - Irreversible blindness in systemic lupus erythematosis. AB - We describe a patient with systemic lupus erythematosis and renal involvement and who developed visual impairment in the absence of hypertension and anti cardiolipin antibodies. This was attributed to occlusive vascular disease and choroidal infarction in the context of active lupus. Complete and irreversible blindness developed rapidly despite treatment and the patient died of cerebral haemorrhage 2 months after discharge. PMID- 8402007 TI - Laser Doppler perfusion imaging: a new technique for measuring skin blood flow in rheumatology. PMID- 8402008 TI - Juvenile ankylosing spondylitis in X-linked agammaglobulinaemia. PMID- 8402009 TI - The abnormal faecal flora of Clostridium perfringens in rheumatoid arthritis and other diseases. PMID- 8402010 TI - Faecal Clostridium perfringens and antibody responses to its antigens in arthritis patients on and off non-steroidal anti-inflammatory drugs. PMID- 8402011 TI - Antineutrophil cytoplasmic antibody titres in patients with recent infection. PMID- 8402012 TI - Small bowel infarction in association with giant cell arteritis. PMID- 8402013 TI - Follow-up of patients with "superficial" transitional cell carcinoma of the bladder: the case for a change in policy. PMID- 8402014 TI - c-myc expression in renal carcinoma: correlation with clinical parameters. AB - There is evidence that c-myc is expressed in renal carcinomas and that it is associated with histological grade. This study reports on the clinical correlations of oncogene expression and clinical features of the disease. Tissue sections from 97 patients (95 primary and 12 secondary cancers) were immunostained with a c-myc oncoprotein antibody. The tumours were scored for extent and intensity of nuclear and cytoplasmic staining. Ninety patients were suitable for follow-up study. High levels of expression correlated significantly with increasing T category and nuclear grade, as well as with venous invasion. No correlation with nodal involvement, the presence of metastases, tumour architecture or cell type was found. The extent of nuclear staining was related to survival (although not to recurrence), but in a multivariate analysis did not provide independent prognostic information. It was concluded that semiquantitative assessment of levels of c-myc oncoprotein does not provide clinically useful prognostic information. PMID- 8402015 TI - Ultrasound in renal carcinoma: an essential in follow-up. AB - A total of 115 patients underwent radical nephrectomy for renal carcinoma in this hospital between 1983 and 1990. Follow-up was available in 113 cases (mean 27 months, range 0-93). The overall actuarial survival was 66% at 2 years and 53% at 5 years. Fifty-three patients have been followed up with annual abdominal ultrasound over the last 2 years. Abnormal ultrasound scans were reported in 6 patients (11%) on follow-up. Two had para-aortic lymph node recurrence amenable to further surgery. One patient developed a metachronous tumour and a partial nephrectomy was carried out. Three patients had local recurrence not amenable to further surgery. Ultrasound and computed tomography have found an invaluable place in the pre-operative diagnosis and staging of renal carcinoma. Sixteen of 113 tumours were diagnosed by ultrasound or intravenous urography carried out for some other purpose; 8 were less than 5 cm in diameter and 12 were T category N0M0V0. The 2-year survival rate in this group was 85% (mean 30 months, range 19 67). We consider the routine follow-up of renal carcinoma with clinical examination and annual chest X-ray to be inadequate. The application of ultrasound in the follow-up of these patients has provided a valuable adjunct to our clinical practice. It has allowed the prompt diagnosis of lymph node recurrence or metachronous tumour amenable to secondary surgery with the potential for improved survival in this group of patients. We consider this to be the logical extension of the value of ultrasound in the pre-operative diagnosis of renal carcinoma. PMID- 8402016 TI - The frusemide test: simple screening test for renal acidification defect in urolithiasis. AB - Urinary pH was first labelled as a risk factor for patients with recurrent calcium stone disease of the renal tract in 1978. The standard diagnostic test for patients with a history of stones, who appear to have urinary acidification defects is the ammonium chloride test. However, the performance of this test results in significant morbidity, particularly nausea and vomiting, and as a result there is little enthusiasm for screening for such defects. We describe a screening test that may be used to determine which patients require more definitive testing. It involves an oral dose of frusemide (40 mg) followed by half-hourly urine sampling for pH. For the detection of a renal tubular acidification defect, the frusemide test had sensitivity = 100%, specificity = 82%, predictive value of a positive result = 40%, predictive value of a negative result = 100%, screening efficiency = 84%, and there was no morbidity. During the ammonium chloride test 80% of the patients vomited or felt very nauseated. PMID- 8402017 TI - Outcome of secondary open surgical procedure in patients who failed primary endopyelotomy. AB - Endopyelotomy is an effective method for treating pelviureteric junction (PUJ) obstruction; however, up to 28% of patients fail endoscopic treatment and require a secondary open surgical procedure. Six patients underwent secondary open surgical procedures following failed endopyelotomy for primary PUJ obstruction at this institution. One patient had impaired renal function in the affected kidney prior to endopyelotomy and postoperative evaluation indicated further deterioration; nephrectomy was performed for chronic obstruction and pain. The remaining 5 patients underwent secondary open pyeloplasty. In 4 patients the operation was difficult and lengthy because of extensive periureteric fibrosis. Difficult dissection was associated with a repeat endoscopic attempt to repair the PUJ obstruction, urinoma formation or stent occlusion requiring intervention. A mean follow-up of 1.7 years demonstrated no radiographic or clinical evidence of recurrent obstruction in the 5 patients undergoing secondary open pyeloplasty; all 6 patients are currently asymptomatic. PMID- 8402018 TI - Ureteroscopy under local anaesthesia with and without intravenous analgesia. AB - In a prospective study of 302 ureteroscopic procedures, 161 were commenced and 133 completed without the use of general or regional anaesthesia. In 15 patients ureteroscopy (URS) was performed with lignocaine jelly in the urethra only, and in 118 with additional intravenous analgesia. Alfentanil, a synthetic morphine derivative, was used for intravenous analgesia. Ureteroscopy was performed prior to extracorporeal shock wave lithotripsy in 46 patients, for stone basketing in 40, for stone fragmentation in 29, for diagnostic purposes in 14 and for cold knife ureterotomy in 4. Ureteric lesions were observed in 9 patients (6.8%) treated under intravenous sedoanalgesia. This percentage is within the range reported in other series of patients treated under general anaesthesia. The findings suggest that URS, when performed without general or regional anaesthesia, does not increase the risk of complications or compromise the results of treatment. PMID- 8402019 TI - Endourological treatment of ureteric injuries. AB - Over the last 8 years, 30 patients with ureteric injuries underwent endoscopic treatment. There were 14 failures, 5 because of blockage or diversion of the catheter in the fistulous gap and 9 because it was impossible to penetrate the stenotic tract. All failures occurred when treatment was attempted more than 3 weeks after the trauma. Sixteen lesions were successfully treated by placing a 6 to 10 F double pigtail catheter in the damaged ureter and leaving it for at least 3 months. Patients were followed up for 24 months. No recurrences were seen and good long-term results were achieved in all cases. In our experience, endourological treatment can be recommended for recent strictures < 2 cm in length, or for small fistulas in which continuity of the ureteric wall is still partially preserved. Despite the risk of failure, especially following late treatment of an injury, it should be considered a safe and effective procedure that is accepted well by the patients and that avoids the need for open surgery and its possible complications. PMID- 8402020 TI - Management of the neuropathic bladder by suprapubic catheterisation. AB - Forty out-patients with neuropathic bladders managed by suprapubic catheterisation have been reviewed. Management included anticholinergic therapy and daily clamping of the catheter to maintain a compliant bladder of good capacity. Twenty-three females and 17 males (mean age 45 years) were studied. Failed intermittent catheterisation and the replacement of long-term urethral catheters were the main indications for suprapubic catheterisation. Catheter related problems were common, with only 5 patients reporting none. Acceptance by patients was high (84%). No evidence could be found of accelerated renal deterioration in patients followed up for more than 2 years. A policy for the management of suprapubic catheters is recommended. PMID- 8402021 TI - Inter-observer variation in assessment of the prostate by digital rectal examination. AB - In a prospective study, inter-observer variation in the assessment of the prostate by digital rectal examination (DRE) by a urologist and a general practitioner was analysed. The 2 physicians performed independent assessment of the prostate in 933 men aged 50 to 69 years with regard to 9 variables as part of a screening programme for carcinoma of the prostate. Complete agreement for all observations was reached in 46.5% of the men. Using kappa (K) statistics, the values were adjusted for the expected chance agreement. K values between 0.485 and 0.682 were obtained for 6 variables representing good agreement; these were size, tenderness, midline furrow, symmetry, induration and nodularity. Agreement regarding fixation, lateral sulci and seminal vesicles was poor owing to the small number of cases deviating from normal. Contrary to the general belief that DRE is highly subjective, our results show a good correspondence between the observations of the examiners when assessing the prostate in a systematic way. PMID- 8402022 TI - Familial transitional cell carcinoma and the Lynch syndrome II. AB - A family is presented in which 4 male siblings developed transitional cell carcinoma (TCC). Four upper tract tumours occurred in 3 and in the fourth the tumour was intravesical. Two of these patients also had colorectal adenocarcinoma. There were 2 other relatives in the pedigree with large bowel cancer. It is suggested that this is an example of Lynch Syndrome II, a hereditary non-polyposis colorectal cancer with extracolonic cancer sites. The implications regarding the screening, surveillance and detection of possible carrier status in healthy relatives is discussed. PMID- 8402023 TI - Coagulation tests in predicting haemorrhage after prostatic resection. AB - It is known that disseminated intravascular coagulation (DIC) can contribute towards blood loss after transurethral resection of the prostate because of the absorption of various prostatic substances. The aim of the present study was to establish whether simple coagulation tests (prothrombin time/activated partial thromboplastin time) carried out immediately after surgery would be useful in predicting those patients who bleed excessively after prostatic resection due to DIC. Criteria to determine significant post-operative haemorrhage were defined. Of 110 patients entered into the study, 34.5% had significant post-operative bleeding and 74% of these had an abnormal prothrombin time (> or = 15 s) immediately after surgery. An abnormal prothrombin time was associated with the resection of large prostates but could also predict the risk of bleeding independent of the resected weight; 18% of patients with an abnormal prothrombin time were also found to have an abnormal activated partial thromboplastin time and all of these had significant blood loss. A group of patients with an abnormal prothrombin time and a resected dry weight > or = 35 g was identified as a high risk group. PMID- 8402024 TI - Prostatic intraepithelial neoplasia and invasive carcinoma in total prostatectomy specimens: distribution, volumes and DNA ploidy. AB - Prostatic intraepithelial neoplasia (PIN) has been postulated to be the main precursor of invasive carcinoma of the prostate (IC). The occurrence, distribution and volumes of PIN and IC in addition to grade were studied in 54 patients who underwent total prostatectomy because of localised IC (T0d-T2 NO MO). PIN always occurred multifocally, localised generally in the peripheral zone (PZ) and was found in all cases. PIN 1 was the most common grade, PIN 3 the least. PIN 3 occurred exclusively in the PZ, in the vicinity of or intermingled with high grade IC. PIN and IC grades were usually concordant. The relative volumes of IC and PIN showed an inverse relationship, i.e. at small IC + PIN volumes PIN dominated, whereas at large IC + PIN volumes both relative and absolute PIN volumes were lower. Furthermore, with increasing PIN grade a tendency towards an increase in tumour volume, Gleason score and frequency of disruption of the basal cell layer was observed. These findings indicate progression from PIN to IC. DNA ploidy of PIN areas was determined by means of flow cytometry. Areas containing PIN 1, 2 or 3 were sampled (1 plug/ml of PIN). All foci displayed only diploid DNA profiles regardless of PIN volume and grade, even with coexistent IC displaying heterogeneous DNA patterns. Our results support the claim that low and medium grade prostatic carcinoma arises from near diploid PIN stemlines and may progress into heterogeneous tumours containing non diploid stemlines. PMID- 8402025 TI - Prognosis of localised prostatic cancer managed by "watch and wait" policy. AB - Progression (within 2-7 years) and cancer-specific mortality (within 4-9 years) were examined in 94 patients with localised (T0-T2 NXM0) prostatic cancer. The patients received no initial anti-cancer treatment. A significant difference was found according to the initial T category both in progression (T0 focal 1/13, T0 diffuse 9/53, T1-2 13/28) and in cancer-related death (T0 focal 0/13, T0 diffuse 3/53, T1-2 6/28). Progression (G1 4/48, G2 17/42, G3 2/4) and cancer-related deaths (G1 1/48, G2 7/42, G3 1/4) also showed significant differences according to histological differentiation. No difference could be demonstrated according to age. PMID- 8402026 TI - Comparison of the effects of prostanoids on human penile circumflex veins and corpus cavernosum tissue. AB - The mechanical effects of the prostaglandins PGE1, PGE2, PGF2 alpha and PGI2 were investigated in human isolated penile circumflex veins (CV) and corpus cavernosum (CC) tissue. PGE1 did not affect resting tension, while the compound produced relaxation of both CC and CV preparations after precontraction with noradrenaline (NA) 3 x 10(-6)M. PGE2 produced contraction in CC and CV preparations. After precontraction with NA, however, relaxation was induced in both tissues. PGF2 alpha induced contraction in CC and CV preparations, while no relaxant responses were seen in preparations precontracted with NA. PGI2 produced no mechanical effects in unstimulated CV preparations, whereas dose-related contraction was induced in CC tissue strips. After precontraction with NA, PGI2 showed no effect in 2 of 6 CC preparations, while relaxation was seen in 4. In CV preparations precontracted with NA, PGI2 produced relaxation. Local synthesis of prostanoids may influence resting tension and modulate NA-induced responses not only in human penile CC but also in CV smooth muscle. PMID- 8402027 TI - The Nesbit-Kelami procedure for congenital curvature of the erect penis. Report on 20 patients. AB - Twenty patients with a congenital deformity of the erect penis were operated upon using the Nesbit-Kelami technique. Excellent results were obtained in all patients. PMID- 8402028 TI - Sonography in fracture of the penis. AB - Sonography was performed in 7 patients with fracture of the penis and in 6 cases it demonstrated the exact site of a tear in the tunica albuginea. It was useful in the diagnosis and management of these patients. PMID- 8402029 TI - Laparoscopic varicocele ligation. AB - A laparoscopic varicocele ligation was performed in 49 men with a clinically proven varicocele. Patients were treated predominantly on a day case or overnight basis with minimal post-operative morbidity. Identification of the testicular artery was aided by a laparoscopic Doppler probe in the majority of patients and preserved in 39 cases. The varicocele persisted in 7 patients, 4 of whom have subsequently had an inguinal exploration confirming a missed testicular vein as the source. In all 7 patients the artery was identified without the use of the Doppler probe, this being suggestive of misidentification of the vessels at laparoscopy. The complications included 4 wound infections, 1 scrotal haematoma and 1 vasal injury which necessitated an open vasovasostomy. Laparoscopic varicocele ligation is a simple and safe technique, causing minimal morbidity and enabling a rapid return to normal activity. The success rate can be improved by accurate identification of the testicular vessels with the use of a laparoscopic Doppler probe. PMID- 8402030 TI - Retroperitoneal lymph node staging of testicular tumours. TNM Study Group. AB - A prospective multicentre study was carried out to determine the efficiency of various diagnostic methods in the assessment of the retroperitoneal space. The diagnostic findings were confirmed histologically after retroperitoneal lymph node dissection (RLND). The sensitivity was 71% for bipedal lymphography, 41% for computed tomography (CT), 31% for abdominal ultrasound and 37% for alpha fetoprotein/human chorionic gonadotrophin (AFP/HCG). Specificity was 60, 94, 87 and 93% respectively. When all diagnostic methods were combined, sensitivity was 88% and specificity 48%. The value of all methods depends on the metastatic enlargement of the lymph nodes. The predictive value of a negative diagnosis was 73% for lymphography, 67% for CT, 61% for ultrasound and 65% for AFP/HCG; the predictive value of a positive diagnosis was 58, 85, 69 and 81% respectively. Despite these results, lymphography is indicated only when a surveillance strategy is planned, since it detected 58% of the lymph node metastases that were overlooked by CT and tumour markers. Despite this, 17% of patients with clinical stage I tumours had metastases. False positive rates are detrimental to primary chemotherapy: between 24% (at least 2 methods positive) and 46% (1 or more methods positive) of patients with clinical stage II A/B tumours had a pathological stage I and for these patients primary chemotherapy meant overtreatment. PMID- 8402031 TI - Childhood urethral injuries: perspectives on outcome and treatment. AB - To elucidate the history of childhood urethral injuries, we undertook a retrospective review of 19 male patients comprising 2 groups: those requiring surgical intervention in adulthood and those with complete disruption of the posterior urethra requiring acute intervention (cystostomy drainage). Group 1 patients (n = 12) ranged in age from 17 to 63 years, had sustained urethral injury between the ages of 3 to 15 years and underwent initial surgery from 9 to 18 years after injury. In all 12 patients the strictures developed in the bulbar urethra. All now report continence and normal voiding and none is impotent. Group 2 patients (n = 7) ranged in age from 4 to 16 years and underwent delayed primary reconstruction of the urethral rupture defect. Six patients are now continent with a good stream but the seventh has required repeat optical urethrotomy. One patient with erectile capability at the time of injury was rendered impotent and remains so. PMID- 8402032 TI - Fungal bezoar causing ureteric obstruction. PMID- 8402033 TI - Formation of a urethral calculus around an unusual foreign body. PMID- 8402034 TI - Tuberculous prostatitis: nodularity may simulate malignancy. PMID- 8402035 TI - Ectopic ureter entering a uterovaginal duplication. PMID- 8402036 TI - Botryoid Wilms' tumour--an unusual variant. PMID- 8402037 TI - Idiopathic testicular microlithiasis: ultrastructural study. PMID- 8402038 TI - Ureteric obstruction due to fat necrosis. PMID- 8402039 TI - Atheroembolism of the prostate. PMID- 8402040 TI - Urostomy specimen of urine--technique of collection. PMID- 8402041 TI - Lithostar view of the mid-ureter. PMID- 8402042 TI - Uncircumcision: a one-stage procedure. PMID- 8402043 TI - Retained balloon catheter. An unusual cause. PMID- 8402044 TI - An unusual "J" stent story. PMID- 8402045 TI - Dartos flap: an aid to urethral reconstruction. PMID- 8402046 TI - Re: Extensive condylomata acuminata of male urethra: management by ventral urethrotomy. PMID- 8402047 TI - Scar formation after breast-conserving surgery for cancer. Sir Charles Gairdner Hospital Breast Cancer Group. AB - A study of surgery for breast cancer was performed to examine the influence of haematoma formation and radiotherapy on the development of mammographically detected scars. Fifty-nine women undergoing lumpectomy and radiotherapy were studied. Sixteen had mammographic scars 12 months after surgery. There was a correlation between postoperative haematoma formation and scarring (P = 0.03) but not between tumour size and scarring. The type of radiation therapy did not influence scar formation. PMID- 8402048 TI - Effects of surgery on the generation of lymphokine-activated killer cells in patients with breast cancer. AB - Natural killer (NK) cell activity and the capacity to generate lymphokine activated killer (LAK) cell activity were studied in 43 patients with operable breast cancer before and after surgery. Results were compared with those from ten healthy subjects. Patients with breast cancer had normal LAK and NK cell activity before surgery. A subgroup of patients with stage III disease had depressed LAK cell activity (P < 0.013). NK cell activity decreased by over 50 per cent on the first day after surgery and did not return to preoperative levels by day 7 (P < 0.0005). Generation of LAK cell activity was unaffected by surgery. The addition of 10 per cent autologous plasma to the culture medium during the induction of LAK cell activity in vitro did not suppress LAK cell activity in patients with breast cancer. These results suggest that perioperative adjuvant immunotherapy based on interleukin 2 and LAK cells is not ruled out by systemic suppressive effects from either cancer or surgery. PMID- 8402049 TI - Tuberculosis in the parotid region. PMID- 8402050 TI - Surgical reintervention for differentiated thyroid cancer. AB - Reoperation was performed in 110 of 185 patients with a differentiated thyroid carcinoma. In 25 patients (23 per cent) the indication for reintervention was a large thyroid remnant and in the other 85 (77 per cent) persistent or recurrent cancer was suspected. In 32 (29 per cent) of the 110 patients undergoing reoperation no evidence of cancer tissue was found. Tumour tissue in 33 patients (30 per cent) was resectable. Of 45 patients (41 per cent) with residual tumour after operation 24 showed only occult thyroid carcinoma with a raised serum thyroglobulin level. Eight of 21 patients with macroscopically persistent tumour died from the disease during a mean follow-up of 2.3 years. In 13 of 38 patients the investigated recurrent tumours were histologically less differentiated than the primary lesions, stressing the importance of total tumour clearance. The treatment of choice for persistent and recurrent differentiated thyroid carcinoma is surgical reintervention, if feasible, before radioiodine and radiation therapy are considered. PMID- 8402051 TI - Small bowel obstruction caused by recurrent cystic lymphangioma. PMID- 8402052 TI - Polytetrafluoroethylene patch grafting for closure of stomach defects in the rat. PMID- 8402053 TI - Prognostic relevance of systematic lymph node dissection in gastric carcinoma. German Gastric Carcinoma Study Group. AB - In a prospective multicentre study of 2394 patients with gastric carcinoma the prognostic relevance of systematic lymph node dissection was evaluated. Of 1654 patients undergoing resection, 558 had a standard lymph node dissection, defined as fewer than 26 nodes in the specimen, and 1096 underwent radical lymphadenectomy, i.e. 26 or more nodes in the specimen. Radical dissection significantly improved the survival rate in patients with Union Internacional Contra la Cancrum (UICC) stages II and IIIA tumours. Multivariate analysis identified radical dissection as an independent prognostic factor in the subgroups of patients with UICC tumour stages II and IIA. Radical dissection conferred no survival advantage in patients with pN2 tumours. There was no significant difference in morbidity and mortality rates between radical and standard lymph node dissection. Radical lymphadenectomy improves survival in patients with UICC gastric cancer stages II and IIIA, and should be the recommended treatment for such patients. PMID- 8402054 TI - Effects of tumour necrosis factor on protein metabolism. AB - Increased skeletal muscle breakdown and negative nitrogen balance are features of sepsis that may be mediated by cytokines. The effects of tumour necrosis factor (TNF) on protein metabolism were studied. When administered to anaesthetized dogs (0.57 x 10(5) units per kg body-weight over 6h), TNF caused urinary nitrogen excretion to increase (mean(s.e.m.) 165(15) mg kg-1 for dogs that received TNF versus 113(8) mg kg-1 for control animals, P < 0.01). Amino acid nitrogen release from the hindlimbs showed no change over the study period, indicating that the additional urinary nitrogen was not derived from peripheral protein stores. In a second study the same dose of TNF or saline was infused after the intestine had been removed. The mean(s.e.m.) urinary nitrogen excretion in control dogs that had undergone enterectomy (101(7) mg kg-1) was similar to that of intact animals, and addition of TNF did not significantly increase nitrogen excretion (86(18) mg kg-1). The results suggest that nitrogen excreted in the urine during administration of TNF is derived, at least initially, from the intestinal tract. PMID- 8402055 TI - Method for diagnosing rejection in small bowel transplantation. AB - Diagnosis of rejection in small bowel transplantation by the identification of a host-cell infiltrate is hampered by the physiological trafficking of host lymphocytes to the 'gut-associated lymphoid tissue' of the graft. This study compared physiological host-cell infiltration of small bowel grafts with that occurring in rejection and stable immunosuppression. Physiological host-cell infiltration, where the graft does not present an immune stimulus to the host, was assessed by transplanting bowel from DA to (DA x PVG) F1 hybrid rats. The extent of host-cell infiltration was determined by immunohistochemical analysis. In the lamina propria, considerable infiltration by host cells was seen, although it was significantly less than that in rejection or stable immunosuppression. By contrast, host cells were seen in the intraepithelial compartment only in rejection. Host-cell infiltration in the absence of an allogeneic stimulus suggests that histological identification of host cells in the lamina propria is not necessarily indicative of rejection. However, the presence of host cells in the intraepithelial compartment is specific for rejection in small bowel transplantation. PMID- 8402056 TI - Surgical polypectomy of duodenal adenomas in familial adenomatous polyposis: experience of two European centres. AB - Between 1978 and 1988, 12 patients with familial adenomatous polyposis (FAP) and severe duodenal polyposis underwent duodenotomy and duodenal polypectomy. The patients were from two separate institutions; all were women, with a mean age of 45.6 (range 25-62) years at the time of duodenotomy. Recurrent duodenal polyposis was detected at endoscopy in all 12 patients, at a mean of 13.3 (range 5-36) months after duodenotomy. All patients had moderate to severe duodenal polyposis at the time of the last endoscopic examination. Local excision of duodenal adenomas is an unsatisfactory treatment option in those with FAP. The ideal management of patients with FAP who have severe duodenal polyposis remains uncertain. PMID- 8402057 TI - Mesenteric infarction: improved survival rate after temporary enterostomy. PMID- 8402058 TI - Incisional hernioplasty with a 'flower' polypropylene prosthesis. PMID- 8402059 TI - Duodenal duplication causing segmental portal hypertension and massive intestinal haemorrhage. PMID- 8402060 TI - Pattern of recurrence after hepatic resection for colorectal metastases. AB - Between 1978 and 1989, 159 patients with liver metastases from colorectal cancer underwent hepatic resection. Of 134 patients in whom metastases were confined to the liver, 109 had tumours removed completely with histologically negative resection margins. Two patients died in hospital. Ultrasonographically guided partial resection was performed in 80 patients, lateral segmentectomy in seven and lobectomy in 22. The 5-year survival rate of patients undergoing potentially curative resection was 47.9 per cent. Patients with metachronous tumours showed a significantly better prognosis than those with synchronous lesions in both univariate (P < 0.01) and multivariate (P = 0.01) analysis. During a median follow-up of 35.4 months, 64 patients developed recurrence, including 34 in the liver, 20 in the lung and 12 in the abdominal cavity. Of those with hepatic recurrence, ten patients developed tumours at the initial resection bed, seven in the same lobe, five in the contralateral lobe and 12 in both lobes. Ultrasonographically guided partial liver resection did not increase the risk of hepatic recurrence. PMID- 8402061 TI - Growth rate and percentage hepatic replacement of colorectal liver metastases. AB - The growth of 32 untreated colorectal liver metastases and hepatic parenchymal volume in 11 patients were studied using planimetry of computed tomographic images. The median percentage hepatic replacement (PHR) of metastases was 0.49 (range 0.09-18.2). At subsequent assessment a median of 72 (range 14-235) days later, metastases were significantly larger with a median PHR of 1.78 (range 0.10 22.9) (P < 0.001). To compare the growth of metastases with variable PHR, logarithmic changes of PHR over 100 days were studied. This parameter was shown to be reduced in metastases of larger volume. The median (interquartile range) non-neoplastic hepatic parenchymal volume was found to increase from 1839 (1647 1997) to 2200 (1590-2401) cm3 (P = 0.005) as metastatic burden increased. These findings suggest that the growth of colorectal hepatic metastases is more complex than a simple process of hepatic replacement. PMID- 8402062 TI - Elective hepatic resection for benign and malignant liver disease: early results. AB - Between August 1989 and April 1992, 60 consecutive elective hepatic resections were performed by one surgeon at two hospitals. This personal series was reviewed to determine the early results of elective hepatic resection. There were 17 patients with liver metastases from colorectal cancer, 14 with hepatocellular carcinoma (three with cirrhosis), seven with cholangiocarcinoma, six with carcinoma of the gallbladder plus liver involvement, ten with liver metastases from other sites and six with benign conditions of the liver. Thirty-eight patients underwent major liver resection, seven unisegmentectomy, six bisegmentectomy, four trisegmentectomy and five non-anatomical resection. Total vascular exclusion was used in 50 cases and the Pringle manoeuvre in ten. The mean(s.d.) operative blood transfusion was 990(1260) ml packed red blood cells (range 0-13 units); 17 patients did not receive blood transfusion. There were two operative deaths; non-fatal complications developed in 16 patients. The two deaths were from postoperative liver failure and there was no other hospital death. PMID- 8402063 TI - Role of serum endotoxin and antiendotoxin core antibody levels in predicting the development of multiple organ failure in acute pancreatitis. AB - It has been proposed that endotoxin contributes to the development of multiple organ failure (MOF) in acute pancreatitis. Endotoxaemia is transient and may not be detected by intermittent blood sampling. By contrast, not only can changes in the patient's endogenous antiendotoxin core antibody pool persist for many days, but depletion of this pool may be a key event in determining the physiological significance of endotoxaemia. A series of 33 patients with acute pancreatitis had daily measurement of Acute Physiology Score (APS) and levels of C-reactive protein, interleukin 6, endotoxin, immunoglobulin (Ig) G and IgM antiendotoxin core antibodies, and prospective documentation of complications. Endotoxin was detected in the serum of 13 patients, while a significant change in levels of endogenous antiendotoxin core antibodies was detected in all those with severe pancreatitis and in 28 overall. MOF developed in seven patients, five of whom died. The combination of a rising APS over the first 48 h of admission and a significant fall in endogenous IgG antibody level was observed in all patients who developed MOF (seven of seven), but in only one of 16 without MOF (P = 0.00003; overall predictive value 91 per cent). This study suggests that measuring the initial trend in APS and the concentration of endogenous IgG antiendotoxin core antibody provides a means of identifying patients with acute severe pancreatitis who are at high risk of developing MOF. This group might benefit from passive immunotherapy with antiendotoxin antibodies. PMID- 8402064 TI - Improved glucose metabolism after subtotal pancreatectomy for pancreatic cancer. AB - Diabetes occurs frequently in patients with pancreatic cancer. To investigate the impact to tumour removal, seven patients were studied before and after 85 per cent subtotal pancreatectomy for adenocarcinoma of the pancreas. The frequency of diabetes was determined by the oral glucose tolerance test. Fasting levels of C peptide and insulin were measured in plasma, and insulin secretion was investigated by hyperglycaemic glucose clamp and glucagon stimulation. Six of the seven patients were diabetic before surgery and four required insulin treatment. Improvements in diabetic status and glucose metabolism were found in all seven patients after operation, as demonstrated by increased glucose metabolic capacity during hyperglycaemia. This occurred despite a postoperative reduction in insulin secretion and is explained by the observed augmentation of whole-body insulin sensitivity after surgery. A diabetogenic factor may be produced by pancreatic adenocarcinoma that may be responsible, directly or indirectly, for the high frequency of diabetes in patients with pancreatic cancer. PMID- 8402065 TI - Complete common bile duct division at laparoscopic cholecystectomy: management by percutaneous drainage and endoscopic stenting. PMID- 8402066 TI - Effect of antitumour necrosis factor treatment on circulating tumour necrosis factor levels and mortality after surgery in jaundiced mice. AB - Tumour necrosis factor (TNF) may play an important role in the increased incidence of complications after surgery in patients with obstructive jaundice. This study evaluated the effect of three different anti-TNF treatments, a monoclonal anti-TNF antibody (TN3), pentoxifylline and lactulose, on outcome after severe surgical trauma in mice with experimental biliary obstruction. Circulating serum TNF levels and mortality rate were monitored. Severe surgical trauma, such as renal ischaemia, in jaundiced mice resulted in increased levels of circulating TNF (3.5 ng/ml) and a high mortality rate (54 per cent). The three different anti-TNF treatments caused a significant reduction in postoperative levels of circulating TNF (TN3, P < 0.001; pentoxifylline, P < 0.01; lactulose, P < 0.05). Treatment with TN3 and pentoxifylline did not lead to a significantly reduced mortality rate (36 and 44 per cent respectively). Only lactulose treatment produced a significantly reduced mortality rate (7 per cent, P < 0.01). TNF is therefore not the only mediator responsible for death after surgery in jaundiced mice. Other mechanisms affected by lactulose are also involved. PMID- 8402067 TI - Probability of rejection predicted from ultrasonographic measurement of renal transplant swelling. AB - Serial ultrasonographic measurements of renal transplant cross-sectional area were used to quantify allograft swelling as a diagnostic test of acute rejection. Eighty consecutive episodes of acute allograft dysfunction (rise in creatinine level > or = 10 per cent or > or = 30 mumol l-1) were investigated. Needle core biopsy was performed in all cases to diagnose acute rejection. Rejecting transplants demonstrated a median (interquartile range) swelling of 16.2 (12.1 25.5) per cent compared with 2.2 (-2.0 to 8.4) per cent for non-rejecting organs (P < 0.001). A > or = 10 per cent increase as the diagnostic threshold for rejection yielded a sensitivity of 80 per cent, specificity of 77 per cent, positive predictive value of 85 per cent, negative predictive value of 71 per cent and overall accuracy of 79 per cent. Using a logistic regression model, predictive probabilities of rejection for individual changes in cross-sectional area were calculated. A 20 per cent increase was associated with a predicted probability of rejection of 87 per cent (95 per cent confidence interval 75-94 per cent). There was a significant correlation between the functional severity of rejection and the degree of transplant swelling (rs = 0.63, P < 0.001). Ultrasonographic measurement of renal transplant cross-sectional area is a simple non-invasive test for the diagnosis of acute rejection. PMID- 8402068 TI - 'Belt loop' stitch. PMID- 8402069 TI - Fistula complicating restorative proctocolectomy. AB - Twenty-seven patients developed a fistula after 168 restorative proctocolectomies. Thirteen fistulas were enterocutaneous (two with communication to the bladder); their origins were from the pouch (three patients), the ileoanal anastomosis (three), the pouch appendage (three), a previous loop ileostomy (two) and iatrogenic small bowel injury (two). Two patients had Crohn's disease. The pouch was removed in four patients, one of whom died from chronic small bowel obstruction; the remaining nine have satisfactory pouch function after fistula excision. Ten pouch-vaginal fistulas occurred, all from the ileoanal anastomosis; four were extrasphincteric. Four of these patients had underlying Crohn's disease. Only two patients, with Crohn's disease and indeterminate colitis, required pouch excision; the remainder have good pouch function after treatment of the fistula. There were three pouch-perineal fistulas, all from the ileoanal anastomosis; these were successfully managed by seton fistulotomy. There was one pouch-vesical fistula, successfully treated by excision of the fistula and pouch appendage. PMID- 8402070 TI - Bile acid absorption after restorative proctocolectomy. PMID- 8402071 TI - Crohn's disease involving a rectus abdominis myocutaneous flap. PMID- 8402072 TI - Relationship between intra-abdominal and intrarectal pressure in the proctometrogram. AB - During normal defaecation the intrarectal pressure increases but, in the absence of a reference catheter in the pelvis, the contribution of abdominal straining and rectal contraction to this rise is unclear. Anorectal manometry was performed in ten consecutive women with no gastrointestinal symptoms in an attempt to measure intrapelvic pressure using a catheter in the bladder. During filling the mean (s.e.m.) rectal pressure increased from 2(1) to 18(4) cmH2O. The mean(s.e.m.) intravesical pressure remained unchanged at 2(1) cmH2O. Evacuation of the rectal balloon produced an increase in mean (s.e.m.) intrarectal pressure from 18(4) (end-filling pressure) to 68(15) cmH2O. The mean (s.e.m.) intravesical pressure increased from 2(1) to 51(18) cmH2O. The true intrarectal pressure (intrarectal minus intravesical) did not rise during defaecation. The rise in intrarectal pressure during rectal evacuation occurs by increased intrapelvic pressure alone. PMID- 8402073 TI - Histological and microbiological assessment of the role of microorganisms in chronic anal fistula. PMID- 8402074 TI - Profiles of workload in general surgery from linked hospital statistics. AB - Data from the Oxford Record Linkage Study between 1976 and 1986 were used to analyse statistical profiles of hospital care, taking account of multiple admissions per patient. Admission rates were higher in the elderly than the young, they were higher for men than women, and they increased over time. Episode based admission rates increased by 15.5 per cent and person-based rates by 11.6 per cent. Most of the increase therefore represented a real rise in the number of people treated rather than an increase in multiple admissions per person. In the latest year of the study there was a mean of 125 admissions per annum per 100 male patients admitted and of 118 admissions per annum per 100 female patients admitted. The mean length of stay per admission and the total time spent in hospital per person per annum declined substantially from 1976 to 1986. Admission rates for prostatectomy, colectomy and varicose vein surgery increased significantly and those for appendicectomy, cholecystectomy and peptic ulcer decreased significantly. PMID- 8402075 TI - Surgical evaluation at the crossroads. PMID- 8402076 TI - Use of an accident and emergency department observation ward in the management of head injury. PMID- 8402077 TI - Gastric cancer and Helicobacter pylori infection. PMID- 8402078 TI - Laparoscopic abdominoperineal resection of the rectum. PMID- 8402079 TI - Laparoscopic truncal vagotomy without drainage. PMID- 8402080 TI - Laparoscopic truncal vagotomy without drainage. PMID- 8402081 TI - Autotransfusion in aortic surgery: the Haemocell 350 cell saver. PMID- 8402082 TI - Heparinized saline to aid aspiration of blood clots during laparoscopy. PMID- 8402083 TI - Clinical trial of tamoxifen in patients with irresectable pancreatic carcinoma. PMID- 8402084 TI - Symptomatic outcome after laparoscopic cholecystectomy. PMID- 8402085 TI - Ultrasonography in the diagnosis of gallbladder polyps. PMID- 8402086 TI - Oxygenation of the gut mucosa. PMID- 8402087 TI - Operative cholangiography and laparoscopic bile duct exploration. PMID- 8402088 TI - Management of superficial femoral artery occlusive disease. AB - The prevalence of intermittent claudication in men aged 55-74 years is 4.5 per cent and a common cause of such claudication is superficial femoral artery (SFA) occlusive disease. The preferred management of patients with this condition remains a subject of discussion. Therapeutic options range from conservative treatment to endovascular intervention and surgical bypass or endarterectomy. Conservative therapy is the primary treatment of choice. However, if this fails and an endovascular technique is chosen, percutaneous transluminal angioplasty (PTA) is the best option; other endovascular methods have failed to achieve higher rates of technical success or patency. PTA should be considered only for short lesions (< or = 10 cm). The usual surgical option for SFA occlusive disease is femoropopliteal bypass using autogenous vein, which has an expected 5-year patency rate of 56-76 per cent. Patency rates decrease if other types of graft are used. An alternative to vein bypass is endarterectomy, with an expected 5 year patency rate of 35-71 per cent. A multicentre randomized trial should be conducted to determine the optimal management of claudication caused by SFA occlusive disease. PMID- 8402089 TI - Surgical evaluation at the crossroads. AB - Surgeons have lagged behind physicians and oncologists in embracing randomized controlled clinical trials. This paper suggests that a complete, accurate and objective comparison of the outcome of a novel intervention with that of a traditional intervention in previous years, or in another surgical group, can yield valuable information and can lead surgeons to improve their practice. There has recently been a decline in the number of randomized controlled trials published in The British Journal of Surgery; this may reflect the unwillingness of many patients to allow their surgical treatment to be decided by chance. PMID- 8402090 TI - Venous assessment using air plethysmography: a comparison with clinical examination, ambulatory venous pressure measurement and duplex scanning. AB - Air plethysmography was compared with clinical assessment, ambulatory venous pressure measurement and duplex ultrasonography in 103 unselected limbs with venous disease and ten normal control limbs without such disease. Measurements of venous function obtained by air plethysmography showed considerable overlap between groups of limbs classified on the basis of clinical condition or by the presence of popliteal incompetence detected by duplex scanning. The measurement of venous refilling time using air plethysmography correlated poorly with that obtained by venous cannulation (rs = 0.58). The residual volume fraction did not correlate with ambulatory venous pressure measurement (rs = 0.04). Air plethysmography was not found to be as useful as previously reported. The residual volume fraction should not be accepted as a substitute for ambulatory venous pressure measurement, which remains the 'gold standard' test of venous function. PMID- 8402091 TI - Aortic anastomosis made easier. PMID- 8402092 TI - Risberg retroperitoneal approach to the abdominal aorta. AB - In a study of the best approach to the infrarenal abdominal aorta, 47 patients were compared retrospectively: 15 underwent a standard transperitoneal incision, 15 a retroperitoneal left flank incision and 17 a new modified lateral pararectus incision, the Risberg approach. Operating time, length of postoperative intubation and hospital stay, mortality rate, morbidity rate and cost were assessed. There was a significant reduction (P < 0.05) in mean(s.d.) operating time (141(21) versus 198(41) min), intraoperative cross-clamping time (74(13) versus 104(46) min) and postoperative intubation time (6.5(8.0) versus 13.3(7.3) h) associated with the Risberg retroperitoneal incision compared with the left flank retroperitoneal route. There was also a significant decrease (P < 0.02) in mean(s.d.) postoperative intubation time (6.5(8.0) versus 17.5(12.0) h), time after operation to discharge (11.0(2.4) versus 17.3(7.6) days) and hospital cost (4885(670) pounds versus 7732(580)) pounds associated with the Risberg incision compared with the transperitoneal approach. The Risberg incision gives better access to the infrarenal abdominal aorta while maintaining the advantages of other retroperitoneal approaches. This technique is recommended as the incision of choice for the retroperitoneal approach to the aorta. PMID- 8402093 TI - Preoperative coagulopathy in ruptured abdominal aortic aneurysm predicts poor outcome. AB - In a prospective study of 50 consecutive patients undergoing operation for ruptured abdominal aortic aneurysm, a coagulation screen was performed on admission to hospital. Twenty patients with either a platelet count < 100 x 10(9)/l or a prothrombin time > 1.5 times the control value had a mortality rate of 65 per cent (95 per cent confidence interval 45-85 per cent); a further 23 patients with normal screen results had a mortality rate of 9 per cent (95 per cent confidence interval 0-20 per cent) (P < 0.001). Seven patients, of whom three died, did not have an admission coagulation screen performed. Patient age in the study group did not have independent statistical predictive power. This study indicates that coagulopathy at the time of admission predicts poor outcome in patients with ruptured aortic aneurysm. Current management strategies are inadequate for the treatment of these patients, who can be rapidly identified on admission by means of platelet and prothrombin counts. PMID- 8402094 TI - Basic fibroblast growth factor accelerates wound healing in chronically ischaemic tissue. AB - The influence of subcutaneously injected recombinant human basic fibroblast growth factor (bFGF) on wound healing in normal (n = 20) and ischaemic (n = 28) skin tissue was investigated. Standardized wounds (5 mm2) were created on the ears of hairless mice and treated for the first 3 days after wound creation with total doses of 720 ng (n = 24) and 4050 ng (n = 24) bFGF. The bFGF had no effect on wound healing in non-ischaemic tissue. In ischaemic skin, mean(s.d.) wound surface area after treatment with 720 ng bFGF was 1.6(0.9), 0.5(0.6) and 0.1(0.3) mm2 compared with 2.8(1.0), 1.4(1.0) and 0.8(0.7) mm2 for control wounds on days 7 (P < 0.04), 10 (P < 0.03) and 13 (P < 0.04) respectively. High-dose bFGF (4050 ng) reduced the mean(s.d.) wound surface area to 2.4(0.7) and 0.8(0.7) mm2 compared with 3.9(0.6) and 2.1(0.8) mm2 for control wounds on days 7 (P < 0.006) and 10 (P < 0.02) respectively. These results suggest that bFGF may be of use for the treatment of wounds in ischaemic tissue. PMID- 8402095 TI - Survival after retroperitoneal necrotizing fasciitis. PMID- 8402096 TI - Computed tomography in the management of blunt abdominal trauma in children. AB - Sixteen children with suspected abdominal injury were reviewed. All underwent plain abdominal radiography and computed tomography (CT). Fifteen CT scans showed an abnormality, and the abdominal radiograph was abnormal in seven cases. Eight patients also underwent abdominal ultrasonography, and six scans failed to demonstrate an abnormality. Only two patients required laparotomy, one for a ruptured left diaphragm and one for a major renal injury. These results support the use of CT as the optimal method for assessing the stable child following blunt abdominal trauma. PMID- 8402097 TI - Transatlantic perspectives of trauma systems. AB - The need for centralized management of acute trauma was evaluated in a 1-year prospective study in Northern Ireland. All patients with an Injury Severity Score > 15 who reached hospital alive were included. The sample population was approximately 1 million people. A total of 239 patients entered the study, of whom 74 died. An audit panel considered that 3-15 per cent of deaths were preventable. There was no significant difference in the preventable mortality rate between any hospital or groups of hospitals. There was a high level of consultant involvement, especially in small hospitals. A system in which patients with acute trauma bypass the nearest hospital to reach a trauma centre may be neither beneficial nor cost-effective in Northern Ireland. Upgrading of the present system with stabilization of the patient and emergency surgery at the nearest hospital before transfer is recommended. PMID- 8402098 TI - Prognostic signs in the evaluation of patients with minor head injury. AB - To assess signs that might be used in the decision whether or not to admit a patient with minor head injury, the records of 713 female and 1163 male patients were reviewed. Skull radiographs were not obtained routinely; all patients were able to walk and talk when they reached medical contact. Nine patients developed an intracranial complication. The risk of developing such a complication was 16.7 per cent when the patient was agitated, 3.4 per cent in the presence of impaired consciousness and 2.1 per cent when positive neurological signs were observed at the time of examination. Based on the medical history, amnesia for > 5 min and vomiting were associated with a risk of 3.3 and 1.2 per cent respectively; the risk increased considerably in the presence of both. It is recommended that all patients presenting themselves with one or more of the above symptoms or signs, or with alcohol intoxication, after a minor head injury be admitted for observation. If these guidelines had been used, all patients with an intracranial complication would have been detected, and 44.5 per cent of the bed-days used would have been saved. PMID- 8402099 TI - Plantar melanoma in black South Africans. AB - The outcome of treatment in 40 black patients (27 women, 13 men; mean age 62.9 years) with plantar melanoma over a 13-year period was analysed to evaluate the efficacy of wide local excision with split skin grafting. Substantial delay in seeking medical attention occurred in 35 patients. At presentation, 20 patients had stage I disease, one stage II, 15 stage III and four stage IV. Acral lentiginous melanoma (27 patients) was the most common histological type. The mean Breslow depth was 6.9 mm and 35 patients had lesions of Clark level IV or V. The mean surface area or plantar lesions was 13.3 cm2. Wide local excision with split skin grafting was used in 29 patients; four patients with neglected advanced plantar lesions had below-knee amputation and seven with metastatic disease did not undergo surgery. Graft sepsis occurred in six patients and local recurrence in two. Nine patients were alive at follow-up; the 5-year survival rate was 25 per cent. Delay in presentation and locally advanced disease may explain the poor prognosis of plantar melanoma in black South Africans. PMID- 8402100 TI - Low-dose tumour necrosis factor alpha and melphalan in hyperthermic isolated limb perfusion. AB - Nine patients with soft tissue tumours of the lower limb not amenable to treatment other than by isolated limb perfusion or amputation underwent hyperthermic isolated limb perfusion at the level of the superficial femoral vessels, using a combination of recombinant tumour necrosis factor (TNF) alpha and melphalan. In seven patients with superficial tumours, necrosis and sloughing was apparent within 48 h of perfusion. All patients experienced a complete tumour response. There were no systemic side-effects associated with the use of TNF alpha although local side-effects, particularly oedema, were pronounced. Three patients ultimately required amputation because of the large soft tissue defects that resulted from necrosis of the tumour and overlying skin. PMID- 8402101 TI - Management of malignant melanoma of the head and neck. AB - A total of 91 patients with melanoma of the head and neck treated between 1973 and 1991 were studied prospectively with regard to prognostic features, treatment and outcome. Other than Breslow thickness, the only important prognostic feature was the less aggressive nature of lentigo maligna melanoma. A policy of selective excision margins, 1 cm for impalpable and 2 cm for palpable lesions, was found to be safe, although preliminary biopsy should be used if there is diagnostic uncertainty or where the expected extent of surgery entails a mutilating procedure. Local recurrence rates were not affected by the method of wound closure, which should be determined by the best functional and cosmetic outcome. These results support the trend against prophylactic neck dissection. Such dissection, when indicated for lesions of the face, pinna, anterior scalp and parotid area, should routinely include superficial parotidectomy. Uncontrolled symptomatic loco-regional recurrence is an uncommon complication that may occur despite radical primary surgery. The role of preoperative radiotherapy for high risk melanoma in this situation warrants investigation. PMID- 8402102 TI - Diathermy in laparoscopic surgery. PMID- 8402103 TI - Silicone gel breast implants, breast cancer and connective tissue disorders. AB - Recent worldwide media speculation that silicone gel-filled breast implants may be linked to an increased incidence of breast and other cancers, and connective tissue disease (particularly systemic sclerosis) has caused concern to the medical profession and public alike. Until carefully controlled studies have been performed to prove the safety of these implants, the US Food and Drug Administration has restricted their use to research and breast reconstruction. Research has so far failed to prove a causative relationship between silicone implants and cancer or connective tissue disorders. PMID- 8402104 TI - Role of surgery in patients with primary non-Hodgkin's lymphoma of the stomach: an old problem revisited. AB - The literature after 1980 dealing with the treatment of primary gastric non Hodgkin's lymphoma (stages I and II) is reviewed. Gastrectomy is recommended for patients with acute symptoms at presentation or with complications of chemotherapy or radiotherapy. In elective cases, patients with stage I disease may be candidates for gastrectomy as a single therapy, provided that tumour-free resection margins are achieved. There is no evidence of substantial benefit from adding postoperative chemotherapy or radiotherapy after an apparently radical resection. In stage II disease, the best results are achieved with ablative surgery plus adjuvant therapy. The resection volume should be limited to macroscopically involved structures, in principle avoiding total gastrectomy and extensive lymph node dissection, since residual disease may be optimally controlled with radiotherapy or chemotherapy. Primary chemotherapy, with or without radiotherapy, represents an experimental approach; the advantages of sparing the stomach must be balanced against the toxicity of an aggressive chemotherapy regimen and the risk of emergency operation for iatrogenic complications. PMID- 8402105 TI - Idiopathic slow-transit constipation. AB - Until recently, the surgical management of idiopathic slow-transit constipation had remained unchanged since the condition was first described by Arbuthnot Lane in 1908. Although colectomy and ileorectal anastomosis is a successful treatment for the majority of patients, symptoms persist or are worse in some cases following such surgery. The previously inaccessible colon is now an area of interest in both health and disease; recent observations on aetiology, clinical investigation, neuropathology and surgical outcome lead us to question the rationale of colectomy for all patients with severe constipation. PMID- 8402106 TI - Acute limb deterioration during intra-arterial thrombolysis. AB - Approximately 12 per cent of limbs undergoing intra-arterial thrombolysis (IAT) develop distal embolism or extension of thrombus during the procedure. These are usually of little clinical consequence and can be treated by increasing the rate of administration of the lytic agent. However, in some patients the clinical condition of the limb deteriorates rapidly. In an attempt to define the incidence of acute limb deterioration during IAT, information was collected from five centres in the UK with experience of the technique. A total of 866 treatments were recorded, with 20 limbs (2.3 per cent) undergoing acute deterioration. This complication was more common during the treatment of thrombosed popliteal aneurysm than during that of emboli or thrombosed atheromatous arteries or grafts (P < 0.001). The amputation rate associated with the complication was high, and operative intervention provided better results than continuation of lysis. PMID- 8402107 TI - Sources of air embolization during carotid surgery: the role of transcranial Doppler ultrasonography. PMID- 8402108 TI - Abdominal aortic aneurysm in the elderly. AB - Thirty-five patients over the age of 80 years underwent surgery for abdominal aortic aneurysm between January 1985 and December 1990. Of these, 25 had surgery for ruptured aneurysm (mortality rate 56 per cent) and ten had surgery for symptomatic or asymptomatic aneurysm (mortality rate zero). The mortality rate for patients with rupture aged over 80 years was considerably greater than that for those under 80 years of age (56 versus 36 per cent). The mortality rate in the elderly population was zero for elective surgery but in those under 80 years it was 5 per cent. During the study period a prospective analysis of all referrals with aortic aneurysm was carried out, whether or not they underwent surgery and whether or not they were admitted to hospital. The ratio of patients accepted for operation to those not accepted for operation was 92:59 (1.56:1) in the under 80-year-old age group and 12:34 (0.35:1) in those aged over 80 years. PMID- 8402109 TI - Systemic leakage during isolated limb perfusion for melanoma. AB - A hazard of regional perfusion for melanoma is incomplete isolation, resulting in leakage of the cytostatic drug into the systemic circulation. Data were analysed retrospectively on 438 melphalan perfusions performed for melanoma of the extremities during the period 1978-1990; continuous isotopic measurement of systemic leakage was carried out. The cumulative systemic leakage after 60 min perfusion was 0.9 per cent (95 per cent confidence interval 0.7-1.1 per cent). Systemic leakage of > or = 1 per cent was detected in 12.6 per cent of perfusions, > or = 5 per cent in 6.2 per cent and > or = 10 per cent in 1.4 per cent. In 2.3 per cent of patients, systemic side-effects in the form of mild transient bone marrow depression occurred. Six variables related to the perfusion technique were assessed by multivariate analysis for their influence on systemic leakage. The level of isolation and diameter of the venous cannula emerged as significant factors. In addition, ligation of the internal iliac vein provided optimal isolation during iliac perfusion. PMID- 8402110 TI - Posterolateral neck dissection: technique and results. AB - Posterolateral neck dissection, alone or combined with a modified radical neck dissection, may be an appropriate method of treating occult and clinically manifest nodal metastases of malignant epithelial tumours of the skin of the posterior scalp. The technique and results are described in 21 patients treated between February 1981 and November 1989. Transformation of nodal status from cN0 to pN+ was seen in five of 13 patients treated electively, who may benefit from such a procedure. During a mean follow-up of 47 months the disease was controlled locally in all patients. Three patients suffered regional recurrence of whom only one had a nodal recurrence within the region previously operated on. The results of this limited retrospective study confirm the value of posterolateral neck dissection in posterior scalp lesions. PMID- 8402111 TI - Staging of supratentorial hemispheric glioma using tumour extension, histopathological grade and extent of surgical resection. AB - A 'TGS' classification for supratentorial brain glioma is proposed, based on three parameters: tumour extension (T), histopathological grade (G) and extent of surgical resection (S). Two groups of patients were analysed: the first (n = 137) had tumour submitted to stereotactic biopsy; the second (n = 60) had malignant glioma operated on by craniotomy and tumor resection. All three staging parameters significantly influenced survival. Histopathological grade was the most significant factor, and the only independent variable on multivariate analysis, followed by extent of tumour and then extent of surgical resection. The data support the applicability of the proposed parameters for a staging classification of hemispheric brain glioma. PMID- 8402112 TI - Graves' ophthalmopathy after subtotal thyroidectomy and radioiodine therapy. AB - Forty-five patients with Graves' ophthalmopathy were studied prospectively. Twenty-one patients (42 orbits; group 1) were treated surgically by subtotal thyroidectomy and 24 (48 orbits; group 2) received radioiodine therapy. All patients underwent tests of thyroid function, ophthalmological examination, and axial and coronal computed tomography before and 12 months after treatment. Patients in group 1 showed significant improvement in the Feldon score (P < 0.001), in proptosis as estimated by Hertel exophthalmometry (P < 0.001) and in proptosis (P < 0.001), maximum diameter of the medial rectus (P < 0.01), lateral rectus (P < 0.05), superior group (P < 0.05) and inferior rectus (P < 0.001) muscles as estimated by computed tomography. Patients in group 2 showed significant improvement only in the Feldon score (P < 0.05). The ophthalmopathy of Graves' disease improves to a greater extent after subtotal thyroidectomy than after radioiodine therapy. PMID- 8402113 TI - Role of computed tomography in the staging of primary breast cancer. PMID- 8402114 TI - Pathophysiology, recognition and management of pneumoretroperitoneum. AB - The medical records of 15 patients found to have pneumoretroperitoneum in a 10 year period were reviewed. The cause of retroperitoneal emphysema was infection in six patients, trauma in five and iatrogenic in four. Emphysema was confined to the retroperitoneum in 11 patients, and extended to the mediastinum in four and to the soft tissue of the neck in three. Failure to diagnose pneumoretroperitoneum resulted in delayed intervention in two patients. One patient with pneumoretroperitoneum and pneumoperitoneum secondary to a lung lesion underwent unnecessary laparotomy. Outcome was favourable in these three patients. The presence of air in the retroperitoneum is not dangerous but its early recognition and detection of the source are important as septic conditions may be involved. PMID- 8402115 TI - Anterior versus posterior reconstruction after transhiatal oesophagectomy: a randomized controlled trial. AB - In a prospective randomized trial the clinical results after transhiatal oesophagectomy with reconstruction in the anterior mediastinum (51 patients) or posterior mediastinum (45 patients) were compared. There were no differences in age, preoperative risk factors, tumour stage and local (surgical) complications between the two groups. However, reconstruction in the posterior mediastinum was associated with significantly fewer days spent in the intensive therapy unit (9 versus 14), fewer cardiopulmonary complications (13 versus 25 per cent) and lower mortality (30-day mortality rate 2 versus 6 per cent; hospital mortality rate 4 versus 10 per cent). These data show superiority of reconstruction in the posterior mediastinum after transhiatal oesophagectomy. This route is strongly recommended, particularly for patients with cardiopulmonary risk factors. PMID- 8402116 TI - Venous thromboprophylaxis in oesophageal cancer surgery. AB - In a retrospective study of 127 patients who underwent resection for oesophageal cancer without anticoagulant thromboprophylaxis, the incidence of clinically established postoperative deep vein thrombosis (DVT) was 1.6 per cent. In a prospective study the Sue-Ling DVT risk factor index was calculated for 53 consecutive patients with oesophageal cancer; the mean (s.d.) value was 4.4(3.5). For 29 patients who underwent resection the following policy of thromboprophylaxis was applied. Patients undergoing transthoracic oesophagectomy with risk score > -4 (four patients) and those undergoing transhiatal resection with risk score > -1 (four) received both anticoagulant and mechanical thromboprophylaxis; the remaining 21 patients received only mechanical thromboprophylaxis. No patient developed DVT after operation. There were no haemorrhagic complications in the group undergoing anticoagulant thromboprophylaxis. The selective approach, based on the Sue-Ling risk factor index and type of operation, spared 72 per cent of the patients operated on from undergoing anticoagulant thromboprophylaxis and produced favourable results in this small series. PMID- 8402117 TI - Abdominal cocoon: a report of two cases. PMID- 8402118 TI - Survival following microscopically confirmed radical resection of N0 gastric cancer. AB - In a series of 128 patients operated on for gastric cancer, 27 satisfied microscopic criteria for radical resection. The median age of these patients was 72.3 (range 36-84) years at operation; the median hospital stay after surgery was 12.9 days. The median observation time was 3.8 years and the crude 5-year survival rate 48 per cent. Older patients did remarkably well. No significant difference was found in the number of survivors, survival-observation time or hospital stay in the age groups 55-64, 65-74 or 75-84 years. Thus, age alone should not be considered a barrier to curative surgical treatment. When the study was ended, 12 of the 27 patients who underwent microscopically confirmed radical resection were still alive; six had died from recurrent gastric cancer and nine from other causes. At 5 years after microscopically confirmed radical resection for gastric carcinoma, the gastric cancer-specific mortality rate was 23 per cent. PMID- 8402119 TI - Extensive versus limited lymph node dissection for gastric cancer: a comparative study of 320 patients. AB - To compare extensive with limited lymph node dissection in the surgical treatment of gastric cancer, 320 patients undergoing gastric resection during 1981-1990 were divided into two groups. Although patients undergoing extended lymphadenectomy (n = 157) had a longer operating time (P = 0.0001) and a greater intraoperative blood transfusion requirement (P = 0.009) than those receiving limited dissection (n = 163), the incidence of postoperative complications (22.3 versus 28.2 per cent, P = 0.13) and the hospital mortality rate (3.8 versus 7.4 per cent, P = 0.12) were similar in the two groups. The 5-year survival rate after curative resection (117 and 121 patients after extensive and limited lymph node dissection respectively) was 65.4 versus 50.1 per cent (P = 0.01): 85.9 versus 82.2 per cent for stage I disease (P = 0.60), 66.1 versus 57.8 per cent for stage II (P = 0.82) and 48.7 versus 29.8 per cent for stage III (P = 0.02). Multivariate analysis using the Cox model showed that the extent of lymphadenectomy was an independent prognostic factor for survival (P = 0.01). The results support the value of extensive lymph node dissection in the surgical treatment of gastric carcinoma. PMID- 8402120 TI - Small bowel prolapse and incarceration caused by a vaginal ring pessary. PMID- 8402121 TI - Factors associated with early discharge after inguinal hernia repair in 500 consecutive unselected patients. French Associations for Surgical Research. AB - The feasibility of discharge within 48 h of surgery was evaluated in 500 consecutive men with unilateral uncomplicated non-recurrent inguinal hernia. Eighty-nine [corrected] patients were unsuitable for short-stay surgery on medical or social grounds. Of 411 patients suitable for early discharge, 107 stayed longer than 48 h. Early discharge was declined by 84 otherwise suitable patients and contraindicated because of local or general complications in 42. A total of 304 patients were discharged within 48 h; 1-day surgery was performed in 51 patients. Employment, low physical requirements, a lower age and fewer than two medical risk factors were associated with feasible and successful short-stay surgery. These factors may not be independent variables. PMID- 8402122 TI - Liver resection with normothermic ischaemia exceeding 1 h. AB - A retrospective study was made of 34 patients who underwent major liver resection with a single period of vascular occlusion exceeding 60 min. The liver remnant was normal in all cases. Vascular occlusion was achieved by continuous portal triad clamping (15 patients), hepatic vascular exclusion (15) or a sequential combination of both procedures (four). Liver cooling was not used. The mean (s.e.m.) duration of continuous normothermic liver ischaemia was 73.6 (2.5) (range 60-127) min. The mean (s.e.m.) amount of blood transfused during operation was 5.3(0.8) units packed red cells. There were no deaths after surgery and the postoperative course was uneventful, or limited to asymptomatic pleural effusion, in 18 patients. Three patients suffered postoperative bleeding necessitating further surgery and one of these required reintervention for a prolonged bile leak. Four patients had transient liver failure that resolved spontaneously within 15 days. There was a 13-fold increase in serum transaminase activities and the proaccelerin level was 45 per cent that of normal on day 1 after operation. These changes were returning to normal levels within 15 days. Continuous vascular occlusion during major liver resection is a useful manoeuvre that may be performed safely on normal hepatic parenchyma for up to 90 min. PMID- 8402123 TI - Computed tomography and fine-needle aspiration cytology for preoperative evaluation of cystic tumours of the pancreas. AB - Cystic neoplasms of the pancreas are rare. They may present as a mucinous (malignant or potentially malignant) tumour or as benign serous cystadenoma. Accurate preoperative diagnosis is therefore essential, as an asymptomatic serous tumour may safely be followed clinically for years, whereas mucinous tumours require aggressive surgery. Thirteen patients with cystadenoma and cystadenocarcinoma of the pancreas were reviewed and how best to improve the rate of preoperative diagnosis was considered. Fine-needle aspiration cytology and computed tomography enable accurate preoperative distinction between mucinous and serous cystic tumours of the pancreas. PMID- 8402124 TI - Morphological studies of graft pancreatitis following pancreas transplantation. AB - Morphological findings in the initial stages of graft pancreatitis were studied systematically in sequential biopsies of 16 human pancreatic allografts. In 14 patients clinical and morphological signs of graft pancreatitis developed in the early postoperative period. In all cases disturbances in the integrity of structures within acinar cells occurred during ischaemia. In ten cases activation of autophagocytosis occurred following reperfusion, with acceleration of cellular metabolism. After reperfusion a marked leucocyte reaction occurred with a later single acinar cell necrosis in six cases. At the same time, high serum pancreatic enzyme concentrations were observed in all patients following transplantation. Exocrine secretion from the allografts via the pancreatic duct was reduced, correlating with the severity of graft pancreatitis. Studies in this clinical situation might complement analyses of the cascade of morphological and pathophysiological reactions during the early stages of other types of acute pancreatitis. PMID- 8402125 TI - Temporal relationship of acinar and microvascular changes in caerulein-induced pancreatitis. AB - A study in rats investigated the temporal relationship between acinar cell changes and alterations in the local microvasculature in oedematous pancreatitis produced by administration of caerulein 5 micrograms kg-1 h-1. Samples were taken from experimental and control animals after 15 min, 30 min, 1 h and 2 h of caerulein infusion. Transmission electron microscopy showed ultrastructural acinar cell changes after 15 min whereas the earliest microvascular changes were seen after 30 min. Ultrastructural alterations in the acinar cells thus preceded local microvascular changes. Microvascular distortion appears to be a consequence and not a cause of pancreatitis in the caerulein model. PMID- 8402126 TI - Resection for adenocarcinoma of the body and tail of the pancreas. AB - Adenocarcinoma of the pancreatic body and tail often presents late and is widely regarded as incurable by surgical resection; long-term survivors are rare. Thirteen patients underwent left resection (n = 7) or total pancreatectomy (n = 6) in a consecutive series of 105 patients with carcinoma of the body or tail of the pancreas. Comparison was made with 17 patients with locally advanced or metastatic disease. Preoperative computed tomography predicted irresectable disease when a large perivascular lymph node mass was demonstrated. Preoperative angiography predicted irresectable disease when there was encasement or obliteration of the coeliac axis or its major branches, or of the superior mesenteric artery or vein. Splenic vessel involvement was sometimes compatible with resection. After resection, median survival was 13 (range 3-50) months, with a minimum follow-up of 2 years. Five patients survived more than 2 years, and three are still alive 30, 43 and 50 months after resection. Resection of carcinoma of the body or tail of the pancreas was possible in 12 per cent of patients and long-term survival was observed in some of these. PMID- 8402127 TI - Laparoscopic cholecystectomy in The Netherlands. AB - Laparoscopic cholecystectomy was introduced into the Netherlands in the Spring of 1990. The aim of this study was to evaluate the results of the procedure in Dutch hospitals over the first 2 years to obtain some insight into its safety and efficacy in general surgical practice. A written questionnaire was sent to all 138 Dutch surgical institutions enquiring about conversion rate, complications (with emphasis on mortality rate and common bile duct injuries), operating time and hospital stay. The surgeons' opinions were also sought on possible contraindications such as previous operation, bile duct stones and cholecystitis, as were their estimations of the percentage of patients in their practice eligible for laparoscopic cholecystectomy. Data were obtained for 6076 laparoscopic cholecystectomies; the response rate was 100 per cent. Conversion to open cholecystectomy was necessary in 413 patients (6.8 per cent), mostly because of adhesions, cholecystitis, haemorrhage and unclear anatomy. Postoperative complications were reported in 260 patients (4.3 per cent). There were seven deaths (0.12 per cent) and 52 (0.86 per cent) bile duct injuries, of which 20 were recognized during laparoscopy. The mean operating time for the ten most recent patients in each institute was 70 (range 30-180) min and the mean hospital stay 4.5 (range 2-8) days. Previous lower abdominal operations were not considered to be a contraindication by 96 per cent of surgeons, whereas previous upper abdominal procedures were regarded as a contraindication by 66 per cent. After successful clearance of the bile duct at endoscopic retrograde cholangiopancreatography, only 12 per cent would perform an open procedure. Moderate cholecystitis was not considered a contraindication to laparoscopic cholecystectomy by 71 per cent of surgeons, but severe cholecystitis was a reason for open cholecystectomy for 83 per cent. In most surgical practices 70-80 per cent of patients were considered to be eligible for the laparoscopic procedure. In conclusion, laparoscopic cholecystectomy has gained rapid acceptance in the Netherlands. Although the number of bile duct injuries is high, the findings of this general survey are similar to those from highly specialized centres and match the overall results of conventional cholecystectomy. PMID- 8402128 TI - New technique of open laparoscopy. PMID- 8402129 TI - Role of early and extensive surgery in the treatment of severe necrotizing soft tissue infection. AB - Forty-two patients with necrotizing soft tissue infection are reviewed. Bacterial culture revealed between two and seven types of micro-organism in each patient. All patients were treated with radical surgical debridement and a combination of antibiotics. In 30 patients, early diagnosis and treatment resulted in only two deaths. Delayed surgical intervention in 12 patients transferred from outside hospitals was followed by nine deaths. Predisposing factors and site of infection did not affect outcome. Of 26 patients with systemic manifestations of sepsis, 16 survivors responded well to initial surgical debridement. PMID- 8402130 TI - Early discharge from hospital after open appendicectomy. AB - A prospective study of 200 consecutive open emergency appendicectomies was carried out. All patients were informed before operation that they would be discharged the following day if possible. The operation was performed through a standard muscle-splitting incision. At the end of the operation the wound was infiltrated with bupivacaine and a diclofenac suppository was given. There were 29 normal, 129 acutely inflamed, six gangrenous and 36 perforated appendices. In all, 147 patients (73.5 per cent) were discharged home within 24 h and reviewed 2 weeks later. Twelve patients had seen their general practitioner, mainly for wound problems (eight) or pain (two). Two others required readmission. Thirty four patients (23.1 per cent) required no postoperative analgesia; 104 (70.7 per cent) required paracetamol or co-proxamol for < or = 3 days and nine (6.1 per cent) for > 3 days. No patient had a problem directly related to early discharge from hospital. Fifty-three patients (26.5 per cent) were not discharged home early due to peritonitis (36), social reasons (seven), nausea (seven), associated caecal volvulus (one) and pregnancy (two). Early discharge from hospital within 24 h after emergency appendicectomy is safe and has good patient acceptability. PMID- 8402131 TI - Synchronous abdominoperineal resection without transfusion. AB - Blood loss and hypovolaemic shock affect the immune system significantly, and perioperative blood transfusion has been shown to be associated with a higher rate of tumour recurrence in patients with cancer and increased susceptibility to infectious complications. Data obtained from patients undergoing synchronous abdominoperineal excision of the rectum were analysed to assess whether such surgery is feasible without transfusion. Twenty-two consecutive unselected patients were studied. There were 16 men and six women of median age 66 (range 48 80) years. The tumour stage was Dukes' B in 11 patients and Dukes' C in 11. Six patients were severely obese according to body mass index. Four patients required blood transfusion (1, 1, 2 and 2 units). There was no hospital mortality and all perineal wounds healed primarily with no wound infection. Fifteen patients received radiation therapy 6 weeks after surgery. Abdominoperineal resection with minimal blood transfusion is feasible in unselected patients. PMID- 8402132 TI - Imipenem prophylaxis in elective colorectal surgery. AB - In a randomized controlled study of 411 patients undergoing elective colorectal surgery, the efficacy and safety of two different regimens of imipenem were compared with those of a control regimen of cefuroxime plus metronidazole. Surgical infections occurred in 92 (26.4 per cent) of 349 evaluable patients. There were no major differences between the three treatment groups. It is concluded that imipenem is as effective as cefuroxime plus metronidazole in the prevention of sepsis after elective colorectal procedures. PMID- 8402133 TI - Effect of putative phospholipase A2 inhibitors on acetic acid-induced acute colitis in the rat. AB - Phospholipase activation may play an important role in ulcerative colitis. This hypothesis was tested by evaluating the effect of two non-selective phospholipase (PL) A2 inhibitors, quinacrine and p-bromophenacyl-bromide (pBPB), on acetic acid induced colitis in the rat. The calcium antagonist verapamil, which may also act as a PLA2 inhibitor, was also tested. Acute colitis was induced in an isolated colonic segment by instillation of 4 per cent acetic acid for 15 s; this induces a uniform colitis after 4 days. The severity of colitis was evaluated histologically, by measuring myeloperoxidase (MPO) activity and by determining plasma exudation into the lumen of the colon (permeability) with 125I-labelled albumin given intravenously. All three putative PLA2 inhibitors tested were found to prevent the development of colitis. Intravenous administration of quinacrine 10 mg kg-1 at 30 min before instillation of acetic acid resulted in a normal mucosal appearance, normal MPO activity and a significantly reduced increase in plasma exudation into the colon. A similar effect was achieved using verapamil. Intracolonic administration of either quinacrine or pBPB also prevented acetic acid-induced colitis. However, three doses, starting immediately after acetic acid administration and repeated on the first and second days, were needed to achieve this, whereas one dose produced only a partial effect. PLA2 may play an important role in acetic acid-induced colitis and inhibition of its activity may offer an alternative mode of treatment in ulcerative colitis. PMID- 8402134 TI - Emergency laparoscopic surgery. PMID- 8402135 TI - Laparoscopic repair of perforated peptic ulcer. PMID- 8402136 TI - Sutureless laparoscopic treatment of perforated duodenal ulcer. PMID- 8402137 TI - Photoplethysmography in the diagnosis of superficial venous valvular incompetence. PMID- 8402138 TI - Photoplethysmography in the diagnosis of superficial venous valvular incompetence. PMID- 8402139 TI - Anaemia and surgery. PMID- 8402140 TI - Abdominal aortic aneurysm: still missing the message. PMID- 8402141 TI - Porcine islet isolation: prospective comparison of automated and manual methods of pancreatic collagenase digestion. PMID- 8402142 TI - Carcinoid tumours of the appendix in children 1957-1986: incidence, treatment and outcome. PMID- 8402143 TI - Laparoscopic herniorrhaphy. PMID- 8402144 TI - Fully implantable venous access systems. PMID- 8402145 TI - Involvement of D2 dopaminergic receptors in the emotional and motivational responses induced by injection of CCK-8 in the posterior part of the rat nucleus accumbens. AB - When CCK-8 was injected in the rat posterior nucleus accumbens, where it is in part co-localized with dopamine, a decrease in exploration of the four hole box and the elevated plus maze was observed. In this study, a selective destruction of the dopaminergic mesoaccumbens pathway induced by local injection of 6 hydroxydopamine (6-OHDA) in the nucleus accumbens was found to suppress the CCK-8 evoked behavioral effects. Moreover, an ex vivo measurement of the dopaminergic metabolism has been performed after injection of CCK-8 in the posterior nucleus accumbens by electrochemical detection of dopamine and its metabolites extracted from punches of brain tissue. The results showed that CCK-8 decreased the turnover of dopamine in the posterior part but not in the anterior part of the nucleus accumbens or in the ventral tegmental area. Furthermore, sulpiride, a selective antagonist for D2 dopamine receptors, but not SCH 23390, a selective antagonist for D1 dopamine receptors, prevented CCK-8-induced behavioral responses. Taken together, these results suggest that CCK-8 could be involved in behavioral adaptation to situations producing change in emotional and/or motivational states through modulation of presynaptic D2 receptor functioning. PMID- 8402146 TI - Suppression of magnetic mu rhythm during parkinsonian tremor. AB - We recorded spontaneous magnetoencephalographic (MEG) activity and somatosensory evoked fields (SEFs) with a 24-channel planar SQUIDgradiometer in five patients with hemiparkinsonism. The SEFs of the patients were within normal limits. During tremorless periods, the spontaneous activity over the somatomotor cortex had a frequency peak at approximately 10 Hz in all five patients and another at approximately 20 Hz in three. Tremor dampened the 10-Hz activity in all patients; in three the effect was bilateral. Tremor did not increase MEG activity at the tremor frequency. The suppression of the mu rhythm by the parkinsonian tremor resembled that occurring during voluntary movements in healthy subjects. PMID- 8402147 TI - Effects of prolonged elevation of potassium on hippocampus of anesthetized rats. AB - We tested the after-effects of prolonged depolarization on neurons in situ in the mammalian brain and examined the site of blockade responsible for failure of synaptic transmission. The CA1 region of the hippocampus of anesthetized rats was exposed to solutions containing elevated concentration of K+ (100-125 mmol/l), administered either by microdialysis in intact brain or by irrigation of the exposed surface of the hippocampus. Recovery was observed for 5-6 1/2 h. When K+ was administered by microdialysis, evoked potentials were recorded from points near (up to 0.2 mm) and far (0.7-1.0 mm) from the dialysis probe. High K+ dialysis induced recurrent waves of spreading depression and, in about half of the preparations, a prolonged unstable depressed state. In the intervals between SD waves orthodromic but not antidromic population spikes remained severely depressed at the 'far' recording site. Following high K+ dialysis orthodromic population spikes recovered in a triphasic cycle: partial recovery with hyper transmission was followed by secondary depression and finally by slow partial or complete recovery. Final recovery was less complete in cases that have experienced prolonged spreading depression. Current source density analysis revealed that during secondary depression transmission was blocked due to failure of dendritic action potentials. When the exposed hippocampus was irrigated with high K+ solution ortho- and antidromic evoked potentials recovered completely following high K+ exposure of less than 30 min, incompletely after 45 min and failed entirely after 60 min. We conclude that prolonged steady depolarization of hippocampal CA1 pyramidal neurons causes lasting loss of function. Dendritic function is especially prone to depolarization-induced injury. CA1 neurons are less vulnerable in situ than they are in vitro. PMID- 8402148 TI - Nickel induces oscillatory behavior and enhanced synaptic and electrotonic transmission between stomatogastric neurons of Panulirus interruptus. AB - The pyloric pattern generator network of the stomatogastric ganglion uses a mixture of burst-inducing plateau potentials, synaptic transmission, and electrical coupling to produce its patterned output. This study examines the effects of two divalent, calcium channel blockers, nickel and cadmium, on voltage oscillations, synaptic transmission, and electrical coupling between the two pyloric dilator (PD) neurons and lateral pyloric (LP) neuron of Panulirus interruptus. The in vitro stomatogastric ganglion was bathed in saline containing tetrodotoxin (TTX) to eliminate Na-spikes and the spontaneous voltage oscillations of the pyloric rhythm, resulting in a steady resting potential. Addition of 50-100 microM Ni2+ to the TTX-saline induced voltage oscillations of similar amplitude and frequency as the endogenous rhythmic activity (before the application of TTX). 25-50 microM nickel enhanced graded synaptic transmission and electrical coupling and altered voltage waveforms, while producing little change in the input resistance measured in the soma. 10-1000 microM Cd2+ acted as a dose-dependent blocker of graded synaptic transmission, but had no other detectable effects. We propose that nickel, in contrast to cadmium, exerts a modulator-like effect deep in the pyloric neuropil. PMID- 8402149 TI - Predator-induced opioid and non-opioid mediated analgesia in young meadow voles: sex differences and developmental changes. AB - The present study examined developmental changes in the nociceptive responses of male and female meadow voles, Microtus pennsylvanicus, exposed to a garter snake, a natural predator of young voles. After 15 min of exposure to the presence of a garter snake, neonatal-juvenile voles (5-20 days of age) displayed naloxone (1.0 mg/kg)-sensitive opioid mediated analgesic responses, while after a brief 30-s exposure to the snake, voles displayed a higher amplitude, non-opioid analgesia that was insensitive to naloxone and blocked by the serotonin-1A (5-HT1A) agonist, 8-hydroxy-2-(di-n-propylamino)tetralin. The levels of opioid and non opioid mediated analgesia declined during development as the threat presented by the snake decreased. Young female voles also displayed a significantly greater non-opioid, 5-HT1A sensitive analgesia than males, with no significant sex differences in the lower amplitude opioid analgesia. These results indicate that young (neonatal) meadow voles that are exposed to a naturally threatening stimulus display sexually dimorphic analgesic responses. These findings also illustrate the need to consider the ecological context when examining environmentally-induced analgesia. PMID- 8402150 TI - Effects of injury discharge on the persistent expression of spinal cord fos-like immunoreactivity produced by sciatic nerve transection in the rat. AB - We recently reported that peripheral nerve injury produced by sciatic nerve transection induces a persistent increase in the expression of the immunoreactive Fos protein product of the c-fos proto-oncogene, an indicator of neuronal activity, in the lumbar spinal cord of the rat and that local anesthetic blockade of the peripheral neuroma attenuates this long-term expression of Fos. In addition to the sustained activity of the injured afferents, the nerve transection itself results, acutely, in a massive injury-induced neural discharge. In this study we evaluated the effect of blocking this massive injury discharge on the persistence of Fos expression. Just prior to nerve transection we applied the short-acting local anesthetic, lidocaine, to the sciatic nerve. Control injections were made subcutaneously on the dorsum of the neck. We report that injection of the local anesthetic, by either route, significantly reduced the number of fos-like immunoreactive neurons at 2 days after nerve transection. The effect was only observed on neurons in the superficial dorsal horn. These results indicate that along with sustained activity of injured afferents and of reorganization of central circuits after injury, the initial brief discharge at the time of nerve injury contributes to a prolonged increase in the activity of spinal cord neurons. PMID- 8402151 TI - Peripheral and central contributions to the persistent expression of spinal cord fos-like immunoreactivity produced by sciatic nerve transection in the rat. AB - Previous studies have demonstrated that noxious stimuli, intense enough to produce tissue injury, evoke a transient expression of the Fos protein product of the c-fos proto-oncogene in neurons, in regions of the spinal cord that contribute to the transmission of nociceptive messages in the rat. Since there is evidence that increases in fos-like immunoreactivity reflect increases in neuronal activity, it has thus been possible to identify populations of neurons that are activated in response to tissue injury. In this study we used immunocytochemical localization of fos-like immunoreactive (FLI) neurons to map the patterns of neuronal activity in the spinal cord at different times after peripheral nerve injury in the rat. Sciatic nerve transection induced a persistent (at least 1 month) elevation in the number of FLI neurons, predominantly in laminae 1, 2, 5, 6 and 7 of the ipsilateral lumbar enlargement of the spinal cord. In the L5 segment, the expression of fos-like immunoreactivity in the superficial dorsal horn (laminae 1 and 2) fluctuated, with peaks of Fos expression at 2 h, 2 days and 2 weeks after nerve transection. Furthermore, by 2 weeks after nerve injury, the distribution of labelled neurons in the superficial laminae of the dorsal horn shifted, with the most densely labelled cells now located in the central portion of the superficial dorsal horn. In contrast, the pattern of labelled neurons in laminae 5, 6 and 7 was relatively constant over the 4-week study period. Local anesthetic block of the sciatic nerve significantly decreased the number of FLI neurons when it was administered at either 2 days or 2 weeks post nerve injury. At 2 days, injection of the local anesthetic subcutaneously in the dorsum of neck, to control for a systemic action, also reduced expression of FLI in laminae 1 and 2; at 2 weeks, the systemic injection of the local anesthetic reduced expression of FLI throughout the gray matter of the spinal cord. These results demonstrate that peripheral nerve injury, in contrast to tissue injury, induces a prolonged increase in Fos expression in neurons predominantly in those regions of the spinal cord that are associated with the transmission of nociceptive messages. This pattern of fos like immunoreactivity is probably the result of persistent neuronal activity in the spinal cord. The increased 'activity' in the spinal cord appears to be maintained both by abnormal activity in the injured peripheral nerve as well as by reorganization of circuits within the spinal cord secondary to the nerve injury. PMID- 8402152 TI - Expression of neuropeptide processing enzymes and neurosecretory proteins in ependyma and choroid plexus epithelium. AB - Recent studies suggest that brain ependyma and choroid plexus produce neuropeptide processing enzymes. To facilitate the understanding of these cells and their ability to produce biologically active peptides, we developed cultures of defined cell type. Ependymal cells were characterized by morphological criteria, and choroid plexus epithelial cell lines were characterized by the presence of the mRNA for IGF-II and transthyretin, a thyroxine binding protein produced in liver and choroid plexus. The ependymal cells and the choroid plexus epithelial cell lines were then examined for the presence of mRNAs for various neuropeptide processing enzymes. Northern blot analysis revealed high levels of furin, carboxypeptidase E, and peptidyl glycine alpha-amidating monooxygenase mRNAs, with levels in ependymal cells comparable to those in brain or pituitary. Carboxypeptidase E activity was detected in medium from cultured ependymal cells; this activity was identified as carboxypeptidase E based on the acidic pH optimum and sensitivity to various inhibitors. The mRNAs for other neuropeptide processing enzymes, such as prohormone convertases 1 and 2, were not detected on Northern blots of RNA from ependyma or choroid plexus epithelium. Since ependyma and choroid plexus epithelium express a subset of processing enzymes, we suggest that these cells have the capacity to produce biologically active peptides. Initial screening by reverse transcriptase-polymerase chain reaction assays has demonstrated the presence of mRNA for the neurosecretory proteins chromogranin B and secretogranin II in both ependyma and choroid plexus epithelium. PMID- 8402153 TI - Effect of guanine nucleotides and temperature on calcitonin gene-related peptide receptor binding sites in brain and peripheral tissues. AB - Recent data have suggested the existence of at least two major classes of calcitonin gene-related peptide (CGRP) receptors in brain and peripheral tissues [Henke et al., Brain Res., 410 (1987) 404-408; Dennis et al., J. Pharmacol. Exp. Ther., 251 (1989) 718-725; ibid, 254 (1990) 123-128; Quirion et al., Ann. NY Acad. Sci., 657 (1992) 88-105]. However, little is currently known in the structure characteristics of CGRP receptors as cloning as yet to be reported. In the present study, the sensitivity of [125I]humanCGRP alpha binding to guanine nucleotides and temperature was investigated in guinea pig atria (prototypical CGRP1 tissue) guinea pig vas deferens (prototypical CGRP2 tissue) and in the rat brain and cerebellum (mixed assay). Binding isotherms of [125I]hCGRP alpha in those four tissue preparations were curvilinear and best fitted to a two-site model under most assay conditions. The high affinity binding component was highly temperature-sensitive and accounted, under experimental conditions, for up to 18% of the total population of receptors. Moreover, these high affinity sites were also highly sensitive to guanine nucleotides (Gpp(NH)p, 100 microM) in all preparations although to a different extend depending upon assay temperatures. Taken together, this suggests that the different CGRP receptor subtypes present in these tissue all belong to a G-protein coupled receptor family. PMID- 8402154 TI - Differing patterns of aberrant neuronal sprouting in Alzheimer's disease with and without Lewy bodies. AB - About one quarter of Alzheimer's disease patients have been found to have concomitant subcortical and neocortical Lewy bodies (LBs). We compared the aberrant neuronal sprouting and the extent of neuritic and synaptic damage in these Lewy body variants of Alzheimer's disease (LBV), with the same pathologic alterations in Alzheimer's disease without LBs (AD). More of the thioflavine-S positive senile plaques of the LBVs contained growth associated protein 43 (GAP 43), a marker of neuritic growth and sprouting. Compared to AD, the LBVs had 39% more GAP-43-positive plaques in the frontal cortex, and 53% more in the hippocampus. These neuritic alterations were accompanied by an accumulation of amyloid precursor protein and phosphorylated neurofilaments. Synapse loss was the same in LBV and AD. These results suggest more extensive aberrant neuronal sprouting in LBV than in AD. PMID- 8402155 TI - Effects of the AMPA/kainate receptor antagonist DNQX in the nucleus accumbens on drug-induced conditioned place preference. AB - Activation of AMPA/kainate glutamatergic receptors in the nucleus accumbens may be a component of the mechanism of drug induced reward. To test this hypothesis, 6,7-dinitroquinoxaline-2,3-dione (DNQX), an alpha-amino-3-hydroxy-5-methyl-4 isoxazole-propionate (AMPA)/kainate glutamatergic receptor anatagonist, was injected into the nucleus accumbens before the administration of amphetamine or morphine during the training phase (acquisition) of a conditioned place preference paradigm. Rats were then tested for place preference in the absence of drugs. In other experiments, DNQX was given before testing for place preference (expression) but not during the training phase. Bilateral injection of DNQX (1 microgram/0.5 microliters/side) inhibited acquisition of place preference to amphetamine (1 mg/kg) but not morphine (10 mg/kg). During acquisition, DNQX marginally attenuated the locomotor stimulation elicited by amphetamine during the first but not subsequent training sessions, while the combination of morphine plus DNQX produced marked akinesia during each training session. When given prior to testing, DNQX inhibited the expression of place preference induced by amphetamine and morphine but did not affect locomotor activity. The results suggest that activation of AMPA/kainate receptors is involved in the primary reward stimulation (acquisition of place preference) of amphetamine but not morphine and in behaviors elicited by memory of primary reward stimulation (expression of place preference) for both drugs. Furthermore, locomotor activity during conditioning is not necessary for acquisition of place preference. PMID- 8402156 TI - Regulation of mouse choroid plexus apical Cl- and K+ channels by serotonin. AB - Using patch-clamp techniques, we have characterized ion channels in the apical membrane of the mouse choroid plexus epithelium and have examined the effect of serotonin on these channels. When the pipette contained 140 mM KCl and the bath contained NaCl Ringer solution, cell-attached patches revealed both Cl- and K+ channels. The Cl- channel was activated by hyperpolarizing membrane potentials, and 70% were also activated by large depolarizing potentials (pipette potential, Vp, more negative than -40 mV). The channel exhibited linear current-voltage (I V) relations with a conductance of 4 +/- 1 pS (n = 30), and a reversal potential at Vp = -14 +/- 1 mV (n = 30). The majority of the K+ channels (84%) were activated by depolarizing membrane potentials. These exhibited linear I-V relations with a conductance of 18 +/- 1 pS (n = 10) and a reversal potential at Vp = -51 +/- 8 mV (n = 10). Serotonin (10(-6) M) increased the open probability (Po) of active Cl- channels (n = 20) by an order of magnitude at the resting potential (Vp = 0 mV) as well as activating previously silent Cl- channels. In contrast, complete inhibition of K+ channel activity was observed in the majority of experiments. There was a 30 s delay after exposure of the tissue to serotonin, thereafter the K+ channel was rapidly inhibited (within 1 min) prior to the stimulation of the Cl- channel. Stimulation of the Cl- channel by serotonin was abolished by mianserin (10(-3) M). We conclude that serotonin exerts its effect on apical Cl- channels via the 5-HT1c receptor. The modulation of these channels by serotonin may be important to CSF secretion and its regulation. PMID- 8402157 TI - Forebrain ischemia in the gerbil increases lambda opiate binding in hippocampal mossy fibers. AB - Transient forebrain ischemia was produced in gerbils by short-term occlusion of the common carotid arteries under halothane anesthesia. Histological analysis of brains 7 days post-ischemia demonstrated characteristic destruction of CA1 pyramidal cells. lambda Opiate binding (measured with [3H]naloxone in the presence of 300 nM diprenorphine) at 7 days post-ischemia was significantly increased in the stratum lucidum of the hippocampus (the mossy fiber layer), but not in any other region measured, including other hippocampal regions, cortex, amygdala, caudate putamen, thalamus, and hypothalamus. The increase in mossy fiber lambda binding was slow to develop (no increase detected up to 48 h post ischemia), and long-lasting (binding remained elevated at 32 days post-ischemia). While MK-801 significantly inhibited CA1 pyramidal cell destruction when administered 20 min prior to ischemia, the increase in mossy fiber lambda binding was still evident. None of seven different opioid agonists and antagonists examined had an effect on either the pyramidal cell damage or increased mossy fiber lambda binding seen 7 days after ischemia. PMID- 8402158 TI - Lack of sensitization or tolerance to the facilitating effect of ventral tegmental area morphine on lateral hypothalamic brain stimulation reward. AB - Unilateral microinjections of morphine (2.5 micrograms/0.5 microliter) into the ventral tegmental area potentiated the rewarding effects of lateral hypothalamic brain stimulation, causing parallel leftward shifts of the functions relating response rate to stimulation frequency. Testing was repeated eight times with no evidence of reliable day-to-day changes either in the direction of sensitization or in the direction of tolerance to the reward-facilitating effects of the drug. The injection dose was sufficient to cause contraversive circling that continued long after the period of reward testing; as with the reward-facilitation, there was neither tolerance nor sensitization to this locomotor effect of the drug. The same dose given to control animals that were not tested for self-stimulation caused progressively stronger circling with repeated testing. Lack of sensitization of the reward-facilitating effects of ventral tegmental morphine raises the possibility of some degree of independence of the brain mechanisms of drug-induced locomotion and of reward-facilitation. PMID- 8402159 TI - Axons and synapses mediating startle-like responses evoked by electrical stimulation of the reticular formation in rats: symmetric and asymmetric collision effects. AB - A new method for determining the locations, directions of transmission and transmission times of synapses mediating electrically evoked responses is proposed here. Electrical stimulation of pontine or medullary reticular formation with one 0.1-ms pulse evokes a short-latency startle-like response. Two pulses were delivered to single sites at various interpulse intervals and the currents required to evoke a criterion startle response were measured. The results suggest that the startle-evoking substrates have absolute refractory periods that range from 0.25-0.6 ms. When one pulse was delivered to a caudal pontine site and a second pulse was delivered to a an ipsilateral medulla site, decreases in required current were observed as interpulse interval increased from +0.4 to +0.8 ms or as interpulse interval decreased from -0.4 to -0.8 ms. These collision-like effects, being symmetric around an interpulse interval of 0, suggest that electrically evoked startle is mediated by fast axons that pass longitudinally through medulla. When one pulse was delivered to the rostral pons and a second pulse to the ipsilateral medulla, however, required currents decreased sharply as interpulse intervals increased from +0.4 to 1.0 ms and as interpulse intervals decreased from +0.2 to -0.2 ms. These asymmetric collision-like effects suggest that strong synapses in the caudal pons, transmitting from pons to medulla, mediate electrically evoked startle. The 0.3-ms asymmetry suggests that the transmission time (i.e., from presynaptic stimulus to postsynaptic action potential) averaged 0.3 ms via monosynaptic connections. The short duration of collision (0.7 ms) suggests that only one postsynaptic action potential was produced with high probability for each presynaptic action potential. From the localization of these effects and the short refractory periods, we estimate that < 60 giant cells on each side of the ventral pontine reticular formation mediate the startle reflex in the rat. PMID- 8402160 TI - Collision-like interactions between acoustic and electrical signals that produce startle reflexes in reticular formation sites. AB - A startle-like response can be evoked at low currents by one-pulse electrical stimulation of reticular formation sites from the rostrolateral pons to the caudomedial medulla. To test whether this response is mediated by the same reticular formation neurons as those that mediate the acoustic startle, we delivered a brief, subthreshold acoustic stimulus followed by an 0.1-ms electrical pulse to one side of the reticular formation of rats. The current thresholds for electrical startle were usually powerfully reduced (50-80%) whenever the acoustic stimulation was presented within 5 ms of the electrical pulse. This summation was, however, interrupted by brief (0.2-1.0 ms) spike-like increases in threshold when the electrical pulse was delivered 4.0-4.6 ms after the offset of the acoustic stimulus. The timing of the spike-like increase in threshold shifted to longer intervals in more caudal sites, consistent with the conduction of action potentials in the startle pathway. For example, the increase occurred at an interval of 4.1 ms near the ventral lateral lemniscus (VLL) and at intervals of 4.4-4.6 ms for sites in the pontine or medullary reticular formation. The increases in startle threshold are attributed to collisions between antidromic action potentials evoked by the electrical pulses and orthodromic action potentials evoked by the acoustic stimuli. These results suggest that the neurons in reticular formation that produce the acoustic startle reflex overlap greatly with the neurons that mediate electrically evoked startle like responses. Also, the acoustic signals mediating the startle reflex must be, in large part, a synchronous volley of action potentials conducted by longitudinal bundles of reticular formation axons. PMID- 8402161 TI - Crossed reticular formation connections that mediate the startle reflex in rats. AB - The startle response is a bilateral response even when elicited by unilateral acoustic or tactile stimuli. Similarly, unilateral electrical stimulation of the reticular formation also elicits a bilateral startle-like response. To examine whether crossed reticular formation connections can distribute the effects of unilateral stimulation across the midline, we delivered one pulse to the caudal pontine (RPC) or medullary reticular formation (MRF) and a second pulse to the opposite side of the brain, at various interpulse intervals. The symmetric collision effects suggest that axons which produce at least 37% (range 23-53%) of the startle response efficacy cross from RPC to RPC with a mean conduction velocity of 13 m/s. Similar collision effects were observed between RPC and MRF sites but at shorter conduction times. To examine which axons might cause these collision effect, the axonally transported label DiI was injected post mortem into 37 RPC sites. Many coarse axons were observed to cross in fascicles between bilateral RPC sites and then separate in the contralateral RPC. The fiber diameters and trajectories of these DiI-labelled axons are consistent with the conduction velocities and trajectories of the substrates mediating the startle like response determined in collision tests. PMID- 8402162 TI - Olfactory bulb organ culture is supported by combined insulin-like growth factor I and basic fibroblast growth factor. AB - A rat olfactory bulb serum-free organ culture system has been developed, using either E19 or day 1 post-natal bulbs. Whereas E19 bulbs were self-sufficient, combined insulin-like growth factor-I (IGF-I) (100 ng/ml) and basic fibroblast growth factor (FGF) (50 ng/ml) supported growth and differentiation of day 1PN bulbs over 6 days. Morphological evidence for viability and differentiation in culture were seen, similar to that occurring in vivo, including development of glomerular layer and maintenance of mitral and tufted and granule cells. Quantitative data derived using radio-labelled amino acid or glucose incorporation supported additive or synergistic effects of IGF-I and FGF. This olfactory bulb organ culture system offers the potential for further studies of brain growth, differentiation and function in vitro. PMID- 8402163 TI - Upregulation of nicotinic receptors following continuous infusion of nicotine is brain-region-specific. AB - Rats receiving 4 mg nicotine/kg/day via implanted minipumps sustained plasma nicotine concentrations of 40 ng/ml throughout two weeks of nicotine infusion. Numbers of brain [3H]nicotine binding sites were increased by about 50% in cortex and hippocampus whereas numbers of [3H]nicotine binding sites in striatum were unaffected by nicotine treatment at either of the timepoints examined (7, 14 days). Cortical [125I] alpha-bungarotoxin and [3H]QNB binding sites were also unchanged. The regional selectivity of nicotinic receptor modulation may reflect the low dose of nicotine used and the mode of administration. The changes observed may be pertinent to the continuous administration of nicotine in man, via transdermal nicotine patches. PMID- 8402164 TI - Cerebral energy metabolism in rats with genetic absence epilepsy is not correlated with the pharmacological increase or suppression of spike-wave discharges. AB - The quantitative [14C]2-deoxyglucose (2-DG) autoradiographic method was applied to measure the effects of pharmacological agents on local cerebral metabolic rates of glucose (LCMRglcs) in a selected strain of Genetic Absence Epilepsy Rats from Strasbourg (GAERS). In a previous study, we have shown that GAERS display an overall significant increase of LCMRglc compared to non-epileptic rats from a selected strain. To further characterize the metabolic responses in GAERS, we measured the effects of drugs aggravating or suppressing absences. The animals were divided into 4 groups, i.e. 2 non-epileptic control groups and 2 GAERS groups. Ten min before the initiation of the 2-DG procedure, both non-epileptic control and epileptic rats received an injection of the same amount of the pharmacological agent, either haloperidol (2 mg/kg) or ethosuximide (200 mg/kg). In the presence of haloperidol, GAERS exhibited almost continuous spike-wave discharges; however, the difference in energy metabolism between GAERS and non epileptic control rats was abolished and LCMRglcs were similar in all structures of both groups of animals. In GAERS treated with ethosuximide, spike-wave discharges were totally suppressed, whereas rates of energy metabolism remained higher by 31-72% in all structures of epileptic rats compared to their corresponding non-epileptic controls. These data demonstrate a lack of correlation between the occurrence of spike-wave discharges and LCMRglcs and are in favor of normal or decreased ictal metabolism and of increased interictal glucose utilization by the brain in rats with absence epilepsy. PMID- 8402165 TI - Permeability and surface area of the blood-nerve barrier in galactose intoxication. AB - The blood-nerve movement of a small molecular weight non-electrolyte was studied in control and galactose-fed rats by measuring the permeability-surface area (PSA) product of the blood-nerve interface to [14C]mannitol in sciatic nerve using an in vivo injection method. PSA products were measured after 9 to 11 months of feeding control rats a diet containing 0% galactose and galactose intoxicated rats a diet containing 40% galactose. Nerves of the galactose-fed group were hydrated as reflected by a significant increase in nerve water content and wet weight to dry weight ratio (both P < 0.05). Compared to controls, PSA products were increased by 51% (P < 0.01) in galactose-fed animals when referenced to nerve dry weight (13.59 +/- 2.90 x 10(-5) ml/s/g dry wt. versus 8.99 +/- 1.59 x 10(-5) ml/s/g dry wt.; mean +/- S.D.; galactose vs. control, respectively) or by 30% (P < 0.001) when referenced to nerve length (2.43 +/- 0.43 x 10(-5) ml/s/mm vs. 1.87 +/- 0.48 x 10(-5) ml/s/mm) but not when referenced to nerve wet weight. It is suggested that in galactose intoxication, where endoneurial volume changes reflect increases in nerve water content, PSA products are best normalized to dry weight or length, which are not affected by volume changes. Normalized to dry weight, the blood-nerve barrier surface area (i.e. vessels and perineurium) was determined by morphometric methods to be increased by 34% in the galactose-intoxicated group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402166 TI - CNS innervation of airway-related parasympathetic preganglionic neurons: a transneuronal labeling study using pseudorabies virus. AB - The CNS cell groups that innervate the tracheal parasympathetic preganglionic neurons were identified by the viral retrograde transneuronal labeling method. Pseudorabies virus (PRV) was injected into the tracheal wall of C8 spinal rats and after 4 days survival, brain tissue sections from these animals were processed for immunohistochemical detection of PRV. Retrogradely labeled parasympathetic preganglionic neurons were seen in three sites in the medulla: the compact portion of the nucleus ambiguus, the area ventral to the nucleus ambiguus, and the rostralmost portion of the medial nucleus tractus solitarius (NTS); this labeling pattern correlated well with the retrograde cell body labeling seen following cholera toxin beta-subunit injections in the tracheal wall. PRV transneuronally labeled neurons were found throughout the CNS with the most abundant labeling concentrated in the ventral medulla oblongata. Labeled neurons were identified along the ventral medullary surface, and in nearby areas including the parapyramidal, retrotrapezoid, gigantocellular and lateral paragigantocellular reticular nuclei as well as the caudal raphe nuclei (raphe pallidus, obscurus, and magnus). Serotonin (5-HT) neurons of the caudal raphe complex (B1-B3 cell groups) and ventromedial medulla were labeled as well as a few C1 adrenergic neurons. The A5 cell group was the major noradrenergic area labeled although a small number of locus coeruleus neurons were also labeled. Several NTS regions contained labeled cells including the commissural, intermediate, medial, central, ventral, and ventrolateral subnuclei. PRV infected neurons were present in the Kolliker-Fuse and Barrington's nuclei. In the rostral mesencephalon, the precommissural nucleus of the dorsal periventricular gray matter was labeled. Labeling was present in the dorsal, lateral and paraventricular hypothalamic nuclei. In summary, the airway parasympathetic preganglionic neurons are innervated predominantly by a network of lower brainstem neurons that lie in the same regions known to be involved in respiratory and cardiovascular regulation. These findings are discussed in relationship to some of the potential CNS mechanisms that may be operative in airway disorders as well as potentially involved in certain fatal respiratory conditions such as Ondine's curse and sudden infant death syndrome (SIDS). PMID- 8402167 TI - Expression of the gene for nerve growth factor (NGF) in the monkey central nervous system. AB - The expression of the gene for nerve growth factor (NGF) was examined in the central nervous system of adult and fetal monkeys. In adults, the highest level of NGF mRNA was found in the hippocampus and relatively high levels were observed in the cerebral cortices and thalamus. NGF mRNA was also detected in the cerebellum and the caudate nucleus. In the spinal cord, there was no evidence of the mRNA. The levels of NGF mRNA were closely correlated with those of NGF. At embryonic day 140 (E140), levels of NGF mRNA in the visual cortex and cerebellum were three times higher than those at the adult stage. Our previous study on the ontogeny of NGF (Hayashi, M. et al., Neuroscience, 36 (1990) 683-689) showed that the level of NGF in the visual cortex at E140 is the same as that at adult stage. Thus, at the fetal stage, NGF may be actively transported from the cerebral cortex to other regions of the brain, such as the basal forebrain area. By contrast, the levels of NGF and NGF mRNA in the cerebellum were almost the same at the adult and fetal stages, suggesting that NGF, which is synthesized in the cerebellum, may be taken up locally by cerebellar cells. PMID- 8402168 TI - Vasoactive intestinal polypeptide immunoreactive structures in the mouse barrel field. AB - Immunohistochemistry for vasoactive intestinal polypeptide was carried out in tangentially cut vibratome sections of the barrel cortex in adult mice. Sections through layer IV have revealed an association between the cytoarchitectonically visible modular organization of barrels and the distribution of immunoreactive axon terminals. These terminals are preferentially localized in the side region of a barrel, whereas the hollow shows a relative scarcity of these structures as shown with image analysis. This finding is the first direct demonstration of a modular distribution of vasoactive intestinal polypeptide-containing axon terminals in the neocortex. PMID- 8402169 TI - Organum vasculosum laminae terminalis (OVLT) is a brain site to produce interleukin-1 beta during fever. AB - The present study was carried out to determine whether interleukin-1 (IL-1) production occurs in the rabbit organum vasculosum laminae terminalis (OVLT) during fever induced by endotoxin. The intravenous (i.v.) injection of endotoxin (4 micrograms/kg) caused significant fever in rabbits. Through the use of in situ hybridization and immunohistochemical techniques, the synthesis of IL-1 was observed in the OVLT during the fever. The present results support the hypothesis that IL-1 is produced in the brain during fever. PMID- 8402170 TI - A possible functional necklace formed by placental antigen X-P2-immunoreactive and intensely acetylcholinesterase-reactive (PAX/IAE) glomerular complexes in the rat olfactory bulb. AB - The relationship between placental antigen X-P2 (PAX)-immunoreactive glomeruli and intensely acetylcholinesterase-reactive (IAE) patchy regions was evaluated by comparison of neighboring cryostat sections of the rat olfactory bulb. Both groups of distribution show similar necklace patterns. Each IAE region consists of heterologous glomerulus-like structures with variable acetylcholinesterase reactivity: strongly and less-reactive (IAE-S and IAE-L) structures. The PAX immunoreactive glomeruli were detected as parts of the IAE-L portions. Three heterologous PAX/IAE glomeruli or glomerulus-like structures (IAE-S, IAE-L/PAX and IAE-L/non-PAX structures) locally form a distinct glomerular complex, the 'PAX/IAE glomerular complex'. At the caudal end of the main olfactory bulb, nine to sixteen such complexes occur at intervals and form a circumferential 'necklace'. Since one of them corresponds to the 'modified glomerular complex' involved in rat suckling behavior, the entire 'necklace' may be associated with processing olfactory stimuli eliciting or suppressing the suckling response. PMID- 8402171 TI - The response of guinea pig auditory-nerve fibers with high spontaneous discharge rates to increments in intensity. AB - We have re-examined the response of auditory-nerve fibres with high spontaneous discharge rates to increments in intensity as a function of the delay of the increment. In agreement with previous studies, the response measured over a relatively long time window (10 ms), emphasising the properties of short-term adaptation, did not decrease with the delay of the increment. However, the response to an increment in intensity, measured over a short time window (0.64 ms), was significantly larger when the increment was coincident with the stimulus onset than when it was delayed by either 5, 10 or 15 ms. PMID- 8402172 TI - Effect of RU 24969 on 5-HT metabolism in the medullary dorsal horn as studied by in vivo voltammetry. AB - The effect of i.p. administration of the preferential 5-HT1B agonist 5-methoxy 3(1,2,3,6-tetrahydro-4-pyridinyl)-1H-indole succinate (RU 24969) (10 mg/kg) has been investigated by in vivo 5-hydroxyindole electrochemical (peak 3) detection in the medullary dorsal horn (MDH) of acute anesthetized and unanesthetized freely moving rats. RU 24969 induced a significant decrease in peak 3 in the MDH of anesthetized rats. In freely moving animals, RU 24969 induced a biphasic effect. Thus, after the injection the curve remained above that of the saline group and returned to control levels up to 60 min. Subsequently the curve decayed to below the control values and rapidly plateaued for up to 180 min. The initial increase and the decrease thereafter were both statistically significant vs. saline. With reference to similar in vivo studies demonstrating the responsiveness of ascending serotonergic systems to RU 24969, it is concluded that the 5-HT metabolism in the serotonergic NMR-dorsal horn system is affected by this 5-HT1B agonist. However, the biphasic effect reported here in unanesthetized animals suggests that RU 24969 could act by two different ways on 5-HT metabolism and indicates that there could be a primary interaction of RU 24969 on the 5-HT uptake system (inhibition) which could, at first, prevail over the interaction with terminal autoreceptors. PMID- 8402173 TI - Stimulation of phosphoinositide hydrolysis by trans-(+/-)-ACPD is greatly enhanced when astrocytes are cultured in a serum-free defined medium. AB - Recent studies have demonstrated that astrocytes have much greater abilities to produce and respond to signalling molecules in the CNS than had been previously estimated. We now report a dramatic enhancement in the ability of a glutamate metabotropic receptor agonist, 1-aminocyclopentane-trans-(+/-)-1,3-dicarboxylic acid (trans-(+/-)-ACPD, to stimulate phosphoinositide hydrolysis in astrocytes cultured in a serum-free defined medium compared with astrocytes cultured in conventional serum-containing medium (43.2 +/- 3.6 vs. 3.2 +/- 0.48-fold of basal, respectively). This enhancement was selective to trans-(+/-)-ACPD as little or no difference in the response to carbachol or norepinephrine was seen between the two culture conditions. These results indicate a great potential for the phosphoinositide pathway in astrocyte glutamatergic signal transduction. PMID- 8402174 TI - Simultaneous visual discrimination learning in lurcher mutant mice. AB - Lurcher mutants were evaluated in the acquisition of a visual discrimination learning task with water escape as a reward. In comparison to normal mice, lurchers were impaired in discrimination learning in a manner not explainable by motor deficits. Although in initial trials the mutants had difficulty in finding the correct invisible platform, in agreement with previous findings of deficits in visuo-spatial organization, they eventually were able to find the quadrants where one of two platforms was situated but even so took longer to reach criterion in choosing the correct one. These results agree with the hypothesis that the cerebellum is involved in the acquisition of visual discrimination tasks in mice. PMID- 8402175 TI - Excitatory amino acid receptors in the nucleus tractus solitarius mediate the responses to the stimulation of cardio-pulmonary vagal afferent C fiber endings. AB - Stimulation of cardio-pulmonary vagal afferent C fiber endings by right atrial injections of phenylbiguanide (PBG, 40 micrograms/kg) elicited apnea, bradycardia and hypotension. These responses were abolished by bilateral vagotomy low in the neck. Stimulation of neurons in a specific nucleus tractus solitarius site (0.5 mm rostral, 0.5 mm lateral and 0.5 mm deep with reference to the calamus scriptorius) by injections of L-glutamate produced responses similar to those following the stimulation of cardio-pulmonary vagal afferent C fiber endings by PBG. Inhibition of neurons in this NTS site by injections of muscimol abolished the responses to PBG. Specific blockade of NMDA receptors by microinjections of AP-7 (100 pmol) or non-NMDA receptors (KA and AMPA) by injections of DNQX (10-25 pmol) into this NTS site did not block the responses to PBG. Microinjections of kynurenic acid (1-4 nmol) into this NTS site blocked the responses to right atrial injections of PBG. These results indicate that: (1) blockade of either NMDA receptors or non-NMDA receptors (KA and AMPA) alone in the above-mentioned NTS site does not abolish the responses to the stimulation of cardio-pulmonary vagal afferent C fiber endings; (2) it is necessary to block NMDA as well as non NMDA receptors in this NTS site for abolishing the responses to the stimulation of these nerve endings. PMID- 8402176 TI - Specific second messengers activate the caudal photoreceptor of crayfish. AB - The caudal photoreceptor (CPR) found in the last abdominal ganglion of crayfish is a well-known example of a non-retinal photosensitive element. In addition to light sensitivity, this cell has been assigned a command role for a walking behavior. The molecular mechanism of transduction in this cell has not been previously studied. The involvement of an intermediate messenger substance is suggested by its long latency to response, its prolonged afterdischarge, and by the requirement for an amplification process for the efficient transduction of light. We tested the effect of some putative second messengers by pressure injecting them into the CPR and noting the physiological response. Here we report that intracellular injection of inositol 1,4,5-trisphosphate (IP3), calcium, and the guanosine nucleotide GTP mimics the light response, while cAMP, IP1 and IP2 have no effect on the firing rate. The key intermediate in transduction in vertebrate photoreceptors, cGMP, was ineffective in this system. This work adds to the growing body of evidence that IP3 plays a role in invertebrate phototransduction. PMID- 8402177 TI - Dopamine transmission increases in the nucleus accumbens of male rats during their first exposure to sexually receptive female rats. AB - In vivo microdialysis was used to monitor extracellular concentrations of dopamine (DA), and its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the nucleus accumbens of sexually naive male rats during their first exposure to sexually receptive or nonreceptive females. DA, DOPAC, and HVA increased progressively and significantly in males that copulated to ejaculation with receptive females. In contrast, DA, DOPAC, and HVA did not increase significantly in males exposed to non-receptive females, despite several attempts by these males to mount the non-receptive females. These results indicate that DA is released unconditionally in the nucleus accumbens of male rats by exposure to sexually receptive female rats, and that copulation with intromission, but not mounting alone, leads to further increases in DA release. PMID- 8402178 TI - Age-related alteration in signal transduction: involvement of the cAMP cascade. AB - In the present study, we have investigated the involvement of the cAMP signal transduction pathways in young and aged rats. A significantly higher endogenous adenosine 3':5'-cyclic monophosphate (cAMP) level and a significant decline of the adenylate cyclase [AC, ATP pyrophosphate-lyase (cyclizing), EC.4.6.1.1.] activity were observed in striatal tissue from young rats (3 months) in comparison to aged rats (approximately 40 months). In the nucleus accumbens (NA), no age-dependent changes in the cAMP concentration and in the AC basal activity were found. To address the question, whether the interactions of guanine nucleotide-binding protein (G-protein) subunits (G alpha s and Gi) with AC have changed in the aging process, various pharmacological agents that modulate the AC activity (e.g., beta, tau-imidoguanine 5'-triphosphate (GppNHp), sodium fluoride (NaF), forskolin (FSK), and the combinations of GppNHp plus FSK, NaF plus FSK, and NaF plus ethanol (ETOH)) were applied. In addition, a [3H]FSK binding test was carried out. In striatal and NA tissue, the stimulation of the AC activity by FSK was inhibited by GppNHp (via Gi-protein) and was superadditive by the combination of FSK and NaF (via Gs-protein). The absolute AC activity upon stimulation by all agents used was significantly lower in the aged striatum compared to young striatum. In the NA, however, the AC activity showed an age dependent reduction only upon FSK and upon FSK plus GppNHp stimulation. There was no difference in the specific [3H]FSK binding to the G alpha s protein-coupled catalytic subunit of the AC between young and aged animals both in the striatum and NA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402179 TI - Psychological stress increases arousal through brain corticotropin-releasing hormone without significant increase in adrenocorticotropin and catecholamine secretion. AB - The effect of psychological and psychophysical stress on pentobarbital (PbNa) induced sleeping time was examined in rats to clarify the influence of psychological stress on arousal. Psychological stress and electric footshock of 5 60 min duration significantly shortened PbNa-induced sleeping time, and the shortening was reversed by intracerebroventricular administration of a corticotropin-releasing hormone (CRH)-receptor antagonist. Electrical footshock and restraint significantly raised plasma adrenocorticotropin (ACTH) and catecholamine levels, whereas psychological stress did not significantly affect the plasma hormones levels. These results suggest that both psychological and psychophysical stress increase arousal through brain CRH. It is also concluded that expression of the central nervous system action of CRH, such as increasing arousal, is not necessarily accompanied by a significant increase in the secretion of ACTH and catecholamine in psychological stress. PMID- 8402180 TI - Functional impairment of hypothalamic corticotropin-releasing factor neurons with immunotargeted toxins enhances food intake induced by neuropeptide Y. AB - Previous work has shown that administration of corticotropin-releasing factor (CRF) into the lateral ventricle antagonizes the orexigenic effect of neuropeptide Y (NPY), and central injection of CRF antagonist, alpha-helical CRF(9-41) enhanced NPY-induced food intake in satiated rats. The aim of the present study was to determine the effects of selective inactivation of hypothalamic CRF neurons on food intake induced by NPY injection and to delineate which hypothalamic nucleus is involved in this NPY/CRF interaction related to the regulation of food intake. Impairment of CRF neuron function by immunotargeting of a ricin A chain toxin with a monoclonal antibody to CRF (CRF-MAb) has been previously reported. Administration of CRF-MAb/toxins into the paraventricular nucleus (PVN) two weeks prior to testing produced markedly enhanced eating induced by injection of NPY into the same nucleus. This effect was accompanied by a 60% decrease in CRF content within the hypothalamus and a 43% decrease of CRF in the median eminence, a site of projection of CRF neurons from the PVN. In contrast, injection of CRF-MAb/toxins into the ventromedial nucleus of the hypothalamus (VMH) did not modify the feeding induced by NPY injection into this hypothalamic area. Systemic pretreatment with the synthetic glucocorticoid dexamethasone at a dose known to downregulate the levels of CRF in the PVN also enhanced the feeding induced by intra-PVN injection of NPY. This suggests that an equilibrium between CRF and NPY neuronal function within the PVN may play an important role in the regulation of food intake. This interactive mechanism may provide some partial explanation of the eating disorders related to stress, in particular anorexia nervosa. PMID- 8402181 TI - Induction of cortical spreading depression with potassium chloride upregulates levels of messenger RNA for glial fibrillary acidic protein in cortex and hippocampus: inhibition by MK-801. AB - The present study evaluates the time course and spatial extent of changes in GFAP mRNA expression following the induction of spreading depression. Spreading depression was elicited by applying filterpaper pledgets soaked in KCl (3 M) to exposed parietal cortex for ten minutes. Animals were killed 1.5, 3, 6, 12, 24, 48, 96 and 192 h post-KCl application, and the forebrains were prepared for quantitative in situ hybridization. The KCl treatment led to a many-fold increase in GFAP mRNA content in the ipsilateral hippocampus and neocortex and, to a lesser extent, in the contralateral hippocampus, but did not affect GFAP mRNA levels in the contralateral cortex or in the thalamus. The time course of increased expression of GFAP mRNA in the hippocampus differed markedly from that of the cortex. In the hippocampus, GFAP mRNA levels rose rapidly to a maximum at 24 h post-exposure, then fell rapidly. In the cortex, levels rose more slowly and did not reach a maximum until 4 days post-exposure. Analysis of GFAP mRNA levels by dot blot hybridization using samples from a separate set of animals killed at one and 4 days following the KCl exposure confirmed both the upregulation in GFAP mRNA levels and the regional time course differences. Intraperitoneal injection of MK-801, a non-competitive NMDA antagonist which prevents spreading depression, blocked the upregulation of GFAP mRNA in both the hippocampus and the cortex, as demonstrated by both in situ and dot blot hybridization. The results suggest that the physiological changes accompanying spreading depression have a powerful influence on glial cell gene expression. PMID- 8402182 TI - Lead increases inositol 1,4,5-trisphosphate levels but does not interfere with calcium transients in primary rat astrocytes. AB - Alteration of receptor-mediated signal transduction pathways by inorganic lead (Pb) has been postulated to contribute to the neurotoxicity of this environmental toxicant, some of these effects involving astrocytes. As Pb is known to mimic Ca2+ in various biological systems or alter Ca(2+)-mediated cellular processes, we analyzed the effect of Pb exposure on alpha 1 receptor activated astrocytic phosphoinositide metabolism and Ca2+ responses in primary astrocyte cultures prepared from cerebral cortex of 1-day-old rats. Exposure to norepinephrine (NE; 10-100 microM) resulted in a significant increase in astrocytic inositol 1,4,5 trisphosphate levels, concomitant with an increase in intracellular Ca2+ levels. Fifteen minute exposure to Pb (10 microM lead acetate) significantly increased inositol 1,4,5-trisphosphate generation compared with controls, both in the presence and absence of NE. However, the inositol 1,4,5-trisphosphate-mediated Ca2+ transients following NE stimulation was unaltered in the presence of Pb (1 100 microM). NE-evoked intracellular Ca2+ responses, both in the presence and absence of extracellular Ca2+ did not differ between control and Pb-treated astrocytes. Additional studies failed to demonstrate the occurrence of Pb influx into astrocytes within the first 12 min of exposure such that Ca2+ responses would be directly affected. It therefore appears unlikely that astrotoxic effects of Pb are mediated via direct changes in intracellular Ca2+ transients. PMID- 8402183 TI - Raphe magnus stimulation-induced antinociception in the cat is associated with release of amino acids as well as serotonin in the lumbar dorsal horn. AB - Stimulation in the nucleus raphe magnus (NRM) inhibits transmission of nociceptive information within the spinal cord through activation of bulbospinal pathways. This study used microdialysis in combination with high pressure liquid chromatography to measure the release of serotonin (5HT) and several amino acids, including glutamate, aspartate and glycine, from the lumbar dorsal horn during electrical stimulation within the NRM in the alpha-chloralose anesthetized cat. Observed release of putative neurotransmitters was correlated with inhibition of nociceptive projection neurons recorded from sites within 800 microns rostral or caudal to the dialysis fiber. NRM stimulus parameters considered to preferentially activate myelinated fibers caused inhibition of nociceptive evoked activity, and increased the release of excitatory amino acids and glycine within the spinal cord, with no detectable release of 5HT. When pulse widths were lengthened and unmyelinated fibers were also activated, increases in 5HT in the spinal dialysate were observed as well. Strychnine administered through the dialysis fiber (0.02-1 mM) antagonized NRM-induced inhibition when 5HT release was not detected. Inhibition produced by stimulation that increased 5HT concentrations was relatively strychnine resistant. These results point to a raphe-spinal inhibitory pathway that is not dependent on 5HT, the activation of which results in the spinal release of glycine. PMID- 8402184 TI - Regional cerebral blood flow following hypothermic circulatory arrest in newborn dogs. AB - A model of hypothermic circulatory arrest has been developed in newborn dogs which simulates the procedure used for the operative repair of congenital cardiac defects in human infants. Hypothermic circulatory arrest for 1.0 h causes no brain damage, whereas cardiac arrest for 1.75 h results in damage of the cerebral cortex, basal ganglia and to a lesser extent the claustrum and amygdaloid nucleus. In the present study, we determined regional cerebral blood flow (rCBF) during 24 h of recovery from hypothermic circulatory arrest. Newborn nitrous oxide anesthetized and artificially ventilated dogs were cooled to 20 degrees C and subjected to cardiac arrest by the i.v. injection of KCl for either 1.0 or 1.75 h. Thereafter, animals were resuscitated, rewarmed to 37 degrees C, and rCBF measured with [14C]iodoantipyrine at either 2 or 18 h of recovery. Control animals were rendered hypothermic to 20 degrees C without cardiac arrest for 1.0 or 1.75 h prior to rewarming. No alterations in CBF at either 2 or 18 h of recovery were present in any of 16 analyzed structures in animals previously subjected to hypothermic circulatory arrest compared to controls rendered hypothermic alone. A direct linear correlation existed between mean arterial blood pressure and blood flow within frontal, parietal and occipital cortex, occipital white matter, hypothalamus and cerebellar vermis in puppies arrested for 1.75 h and recovered for 2 h, suggesting a loss of CBF autoregulation at this interval. No such association between blood pressure and CBF was apparent at 18 h of recovery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402185 TI - Motor learning deficits in aged rats are correlated with loss of cerebellar noradrenergic function. AB - We have demonstrated that aged rats show impairments in learning patterned motor movements. Similar behavioral impairment is observed in rats with noradrenergic lesions. Norepinephrine is known to act as a neuromodulator in the cerebellar cortex because it can augment the action of GABA and other neurotransmitters. This effect of NE to augment the signal to noise ratio of GABAergic inputs to cerebellar Purkinje neurons is a possible substrate for NE's effect on motor learning. Aged rats demonstrate deficits in the modulatory actions of NE to augment GABAergic inhibitions when both substances are locally applied onto cerebellar Purkinje neurons. In this report, we examined how motor learning and cerebellar noradrenergic function varied in individual young and 20-month-old Fischer 344 rats. There was a significant correlation between the loss of the neuromodulatory actions of norepinephrine (NE) in the cerebellar cortex and the rate of learning a novel motor task in individual rats. This report thus demonstrates for the first time a correlation between age-related impairments in motor plasticity and specific neurophysiological deficits in cerebellar Purkinje neurons in individual animals. PMID- 8402186 TI - Electroconvulsive stimulation and synaptic plasticity in the rat. AB - The effects of repeated, spaced, electroconvulsive stimulation (ECS) on rodent hippocampal synaptic plasticity was investigated in vivo. Long-term potentiation (LTP) was induced in intact rats using high-frequency perforant path stimulation, and measured by recording extracellular excitatory field potentials and population spikes evoked in the dentate gyrus by low-frequency stimulation before and after LTP induction. LTP induction appeared to be inhibited in animals which had received ECS. However, inspection and analysis of absolute excitatory postsynaptic potential and population spike size before LTP induction in ECS treated animals suggested that LTP may have already been induced as a consequence of seizure activity, reducing the degree to which further potentiation could be elicited experimentally. PMID- 8402187 TI - Localization of nitric oxide-related substances in the peripheral nervous tissues. AB - Nitric oxide (NO) is now recognized as a transduction molecule in many biological systems, and is known to promote the synthesis of cGMP by activating the soluble guanylate cyclase. NO synthase which fully accounts for all the neuronal activity of NADPH diaphorase catalyzes L-arginine to NO and L-citrulline. In the present study, the localization of NO-related substances, L-arginine, NO synthase, L citrulline and cGMP in the enteric plexus and dorsal root ganglia was demonstrated with immuno- or enzyme-histochemical methods. L-Arginine was proved accumulated in glial cells, while NO synthase and L-citrulline were found in neurons. Cyclic GMP was predominantly observed in glial cells. These results reveal L-arginine-NO-cGMP pathway may be present in the enteric plexus and dorsal root ganglion as in the brain, and provide visible evidence that NO mediates neuron-glia communications in this pathway. PMID- 8402188 TI - Neurogenetical segregation of the vestibulospinal neurons in the rat. AB - The time of origin of the vestibulospinal projection neurons was determined by a double-labeling method using 5-bromodeoxyuridine (BrdU), the thymidine analogue, and Fluoro-Gold (FG), a retrograde fluorescent tracer. Rat fetuses were exposed to BrdU in utero to label the vestibular neurons on one of the embryonic (E) days between E12 and E15. Upon reaching adulthood, the rats were given unilateral injections of FG into the cervical cord to identify the spinal projection neurons. Brainstem sections were immunohistochemically processed for BrdU and then examined for neurons that were both BrdU-positive and FG-positive in the vestibular nuclei. In the lateral vestibular nucleus (LVe), most of the vestibulospinal neurons were generated on E12. In the inferior vestibular nucleus (IVe), the vestibulospinal neurons were produced almost equally on both E12 and E13. In the medial vestibular nucleus (MVe), the vestibulospinal neurons were generated consistently on days between E12 and E14 with a mild peak on E13. The present results thus demonstrate that genesis of the vestibulospinal neurons occurs sequentially in the following order: firstly in the LVe, secondly in the IVe, and finally in the MVe. The different sequential generation of vestibulospinal neurons among the LVe, MVe and IVe may reflect the fact that the vestibulospinal projections are differentially organized depending on the nature of each subnucleus. PMID- 8402189 TI - Immunocytochemical localization of enkephalin in the cat visual cortex. AB - The localization of enkephalin-immunoreactivity in the cat visual cortex (area 17) was analyzed by using immunohistochemical methods with a monoclonal antibody directed against enkephalin. The majority of the immunoreactive product was localized in neuronal processes. The density of immunopositive fibers was greatest in layer VI, with moderate staining in layers I, II, III and V, and the least dense staining in layer IV. Layer IVab neurons showed a striking concentration of immunopositive puncta around their cell bodies. Immunopositive neurons were scarcely present in the visual cortex. They were found in all cortical layers, but mostly in layer VI. The immunopositive neurons were non pyramidal, mostly multipolar in shape and occasionally bipolar. The results provide anatomical evidence that enkephalin may have modulatory effects on visual cortical neurons. PMID- 8402190 TI - Repeated naloxone treatments and exposures to weak 60-Hz magnetic fields have 'analgesic' effects in snails. AB - Results of studies with rodents have shown that animals repeatedly injected with the opioid antagonist, naloxone, acquire a hypoalgesic response to thermal nociceptive stimuli. The present study revealed a similar response in the terrestrial pulmonate snail, Cepaea nemoralis. Snails receiving daily injections of naloxone followed by measurements of thermal nociceptive sensitivity also developed hypoalgesia. Daily brief (30-min) exposures to a weak 60-Hz magnetic field (1.0 gauss or 0.1 mT), which acutely antagonize opioid-mediated nociception and antinociception in a manner comparable to that of naloxone, also led to the expression of a hypoalgesic responses. This suggests that opioid antagonist induced thermal hypoalgesia may be a basic feature of opioid systems. This naloxone- and magnetic field-induced 'analgesia' is consistent with either a facilitation of aversive thermal conditioning and or antagonism of the excitatory, hyperalgesic effects of low levels of endogenous opioids. PMID- 8402191 TI - A novel cholinergic-specific antigen (Chol-2) in mammalian brain. AB - Three new antisera have been raised in sheep against cholinergic electromotor presynaptic plasma membranes prepared from the electric organs of the electric ray, Torpedo marmorata. They all recognized one or more cholinergic-specific antigens in the mammalian nervous system by the following criteria: they sensitized the cholinergic subpopulation of rat-brain synaptosomes--and only this subpopulation--to lysis by the complement system and, in an immunocytochemical study, selectively stained choline acetyltransferase-positive cholinergic neurons in the rat spinal cord. However, two of the three antisera failed to recognize Chol-1 alpha and -beta, two closely related minor gangliosides already identified as the cholinergic-specific antigens recognized by previous anti-Torpedo presynaptic plasma membrane antisera or indeed any other ganglioside and the third recognized only Chol-1 alpha. A further investigation of the antigen(s) recognized by the most antigenic of the new antisera indicated that it is proteinaceous in nature, but has epitopes in common with electric organ gangliosides. PMID- 8402192 TI - Effect of aging in human cortical pre- and postsynaptic serotonin binding sites. AB - 5-HT1A, 5-HT1D, 5-HT2 binding sites and affinity and 5-HT uptake sites were simultaneously determined in frontal cortex samples from 23 control subjects, aged 16-75 years. A significant reduction in the number of 5-HT1D and 5-HT2 binding sites was found with regard to age, together with a significant decrease in the 5-HT2 binding affinity. It is suggested that the total 5-HT1 age-related loss described in previous studies could be ascribed to the 5-HT2 subtype. Furthermore, aging does not seem to be associated with a reduced cortical serotonergic innervation, as indicated by the stability of the [3H]paroxetine labeled 5-HT uptake sites. PMID- 8402193 TI - Sex differences in NMDA receptor mediated responses in rats. AB - The behavioural effects of the N-methyl-D-aspartate (NMDA) receptor antagonist MK 801 (dizocilpine) were compared in age-matched female and male Wistar rats. At a dose of 0.1 mg/kg i.p., almost no behavioural alterations were seen in male rats, while marked and long-lasting phencyclidine-like behavioural alterations, such as ataxia, hyperlocomotion, and head weaving were observed in female rats. When the dosage of MK-801 was increased to 0.3 mg/kg, these behavioural alterations were also observed in male animals, but with lower intensity and/or duration than in female rats. The data demonstrate that female rats are much more susceptible to NMDA receptor blockade than males, suggesting sex related differences in the endogenous modulation of this glutamate subreceptor. PMID- 8402194 TI - Increase in ubiquitin conjugates dependent on ischemic damage. AB - Insoluble ubiquitin conjugates (UC) in the mitochondrial fraction of the gerbil cortex were analyzed following transient forebrain ischemia. At 1 h of reperfusion after 2-10 min of ischemia, UC increased as the duration of ischemia was prolonged. Pre-treatment with pentobarbital, rather than post-treatment immediately after recirculation, reduced the increase of UC at 1 h of reperfusion following 5 min of ischemia. Pentobarbital had no effect on in vitro ubiquitination of heat-denatured lysozyme by the extract of gerbil cortex. These results suggest that increase in UC is dependent on ischemic damage and pentobarbital attenuates the increase of UC by relieving injury during ischemia. PMID- 8402195 TI - Alterations of guanine nucleotide-binding proteins in post-mortem human brain in alcoholics. AB - Qualitative and quantitative alterations of G proteins in membrane preparations from parietal and temporal cortex regions in post-mortem brains obtained from alcoholics and controls matched with respect to age and post-mortem delay were investigated by Western-blotting with polyclonal antibodies against specific G protein subunits and functional photoaffinity GTP binding. Quantitative immunoblotting showed that only Gs alpha (52 kDa species) in temporal cortex was significantly decreased (30%, P < 0.05) in alcoholics compared with controls. Moreover, ethanol-stimulated photoaffinity GTP labeling of Gs alpha and Gi/o alpha was decreased in alcoholics in both cortex regions. These results suggest that disturbances of G protein-mediated signal transduction may be involved in the pathophysiology of alcoholics. PMID- 8402196 TI - Prevention by eliprodil (SL 82.0715) of traumatic brain damage in the rat. Existence of a large (18 h) therapeutic window. AB - The neuroprotective potential of eliprodil (SL 82.0715), an N-methyl-D-aspartate (NMDA) receptor antagonist acting at the polyamine modulatory site, in brain trauma was examined in a rat model of lateral fluid-percussion brain injury. The volume of the lesion was assessed histologically by measuring, at 7 days post injury, the area of brain damage at 10 coronal planes. Eliprodil (10 mg/kg i.p.) when given 15 min, 6 h and 24 h after fluid percussion (1.6 atm) and then b.i.d. for the following 6 days, reduced by 60% the volume of cortical damage. A similar neuroprotection was obtained when the first administration of eliprodil was delayed by up to 12 h after the brain insult. Moreover, when the treatment with this compound was started at 18 h post-injury, cortical damage was still significantly reduced by 33%. Autoradiographic studies showed that eliprodil treatment (10 mg/kg, i.p.), initiated 15 min after the trauma, also caused a marked reduction of the loss of the neuronal marker omega 1-2 (central benzodiazepine) binding sites and of the increase in the glial/macrophage marker peripheral type benzodiazepine binding sites in the cerebral cortex. In contrast, dizocilpine (a blocker of the cationic channel coupled to the NMDA receptor) when administered 6 h and 24 h after fluid percussion and then b.i.d. for the following 6 days induced a non significant reduction of the volume of the lesion at the highest tolerated dose (0.6 mg/kg i.p.). These results demonstrate the neuroprotective activity of eliprodil in experimental brain trauma using neuropathology as an endpoint and indicate that there is a very large time window for therapeutic intervention, consistent with the delayed nature of the neuronal loss, in this condition. PMID- 8402197 TI - Muscarinic and nicotinic receptor changes in the cortex and thalamus of brains of chronic alcoholics. AB - The cholinergic system was studied in the cortical and thalamic brain tissues obtained at autopsy from 21 chronic alcoholics and 20 controls. The age related decrease in choline acetyltransferase (ChAT) activity observed in the thalamus of control brains was not found in the corresponding brain areas of chronic alcoholics. A significant decrease in the number of muscarinic receptor binding sites was observed with age in the frontal cortex of both controls and chronic alcoholics when analysed with the nonselective muscarinic antagonist [3H]quinuclidinyl benzilate ([3H]QNB). A significant increase in the number of muscarinic receptor binding sites was observed in the thalamus of controls but not in chronic alcoholics. When the subjects were divided into young (19-57) years, and old (59-84 years) chronic alcoholics marked losses in the total number of muscarinic receptors as well as M1 and M2 receptor subtypes were found in the thalamus of the old group of alcoholics compared to age-matched controls. A coupling of muscarinic receptors to G proteins was observed in thalamic tissues from both controls and chronic alcoholics. Guanylyl-imidodiphosphate (Gpp(NH)p) induced a steepening and rightward shift of the carbachol/[3H]QNB displacement curves performed in membrane preparations of the thalamus from both controls and chronic alcoholics. The number of high affinity nicotinic binding sites in the frontal cortex and thalamus did not differ significantly between controls and chronic alcoholics. PMID- 8402198 TI - Dipeptidyl peptidase II in astrocytes of the rat brain. Meningeal cells increase enzymic activity in cultivated astrocytes. AB - Astrocytes grown in media conditioned by meningeal cells (MCM) develop cellular processes and markedly increased protein per cell. One protein component affected is the dipeptidyl peptidase II (DPP II). The increase of DPP II activity is dose- and time-dependent and can also be elicited by the second messenger cAMP. More mature astrocytes express higher levels of DPP II than immature proliferating astrocytes. The rate of proliferation of astrocytes is markedly enhanced by enriched MCM. These observations lead to the assumption that DPP II has a function within the catabolic processes of cellular differentiation. To assess whether the in vitro results may reflect in vivo conditions, we investigated the postnatal development of DPP II in the rat brain. Differentiating astrocytes in vivo are especially found early postnatally and, indeed, during this period high specific activities are found in brain. Depending on the region investigated DPP II activities decrease within the first ten days to one fourth of their P2 level and finally reach at about similar levels in all brain regions. Exceptions are the hypothalamus, where the activity is generally 1.5- to 3-fold higher than elsewhere in brain, and pons and mesencephalon, where the perinatal activity peak is lacking. The bulk activity of DPP II in immature rat brains is attributed to differentiating astrocytes loosing it in later postnatal stages due to a neuronal influence. PMID- 8402199 TI - Rolipram enhances the development of voltage-dependent Ca2+ current and serotonin induced current in rat pheochromocytoma cells. AB - The effects of chronic treatment (6-8 days) with a phosphodiesterase inhibitor, rolipram, on the expression of voltage-dependent Ca2+ channels, nicotinic acetylcholine (ACh) receptors and 5-hydroxytryptamine (5-HT) receptors were investigated in PC12 cells. The results were compared with the effects of nerve growth factor (NGF), 8-bromo-cyclic AMP (8-Br-cAMP) and phorbol 12-myristate 13 acetate (PMA). In the morphological study rolipram, at a high concentration (100 microM) induced the extension of neurites. A similar result was obtained in 8-Br cAMP (1 mM)-treated cells. Rolipram, at a low concentration (10 microM) or PMA (10(-7) M) did not induce obvious morphological change. NGF (100 ng/ml) induced the extension of long neurites and the formation of neural networks. Rolipram (100 microM) increased the current density (pA/pF) of voltage dependent Ca2+ current (ICa). Both NGF and 8-Br-cAMP also increased the current density of ICa, whereas PMA did not. NGF increased the current density of the nicotinic ACh response whereas rolipram, 8-Br-cAMP and PMA decreased. Rolipram (100 microM), NGF (100 ng/ml), and 8-Br-cAMP (1 mM) increased the current density of the 5-HT response whereas the effect of PMA (100 nM) was slight. The results suggest that rolipram is able to contribute to the neuronal development by increasing intracellular cAMP as well as 8-Br-cAMP. Consequently, rolipram behaves like a neurotrophic factor in cultured PC12 cells. PMID- 8402200 TI - The endogenous lectin RL-29 is transynaptically induced in dorsal horn neurons following peripheral neuropathy in the rat. AB - Neurons containing immunoreactivity to the endogenous lactose-binding lectin, RL 29, were examined in the L4 segment of the spinal cord of rats with an experimental neuropathy. The cells appeared by 5 days and were also present at 14 and 28 days postoperatively. All neurons were found in the dorsal horn ipsilateral to the injury. The neurons were multipolar and the reaction product revealed the morphology of the primary and secondary dendrites and some axons. Most of the neurons were located in the reticulated region of the dorsal horn, corresponding to Rexed's lamina V. In 14-day neuropathy animals treated with the NMDA-receptor antagonist MK-801, the number of RL-29 cell profiles observed was significantly reduced. Double labeling experiments revealed that spinothalamic tract neurons did not contain RL-29. The results suggest that the neuropathic injury produces a long term, transynaptic change in a subpopulation of dorsal horn neurons, that is mediated by excitatory amino acid transmitters acting at NMDA receptors. PMID- 8402201 TI - Effects of the memory enhancer linopirdine (Dup 996) on cerebral glucose metabolism in naive and hypoxia-exposed rats. AB - Linopirdine [DuP 996; 3,3-bis(4-pyrindinylmethyl)-1-phenylindolin-2-one] represents a novel class of compounds which enhance depolarization-activated (but not basal) release of acetylcholine, dopamine and serotonin in brain slices and improve learning and memory in rodents. The effects of linopiridine on local cerebral glucose metabolism were studied by the quantitative autoradiographic 2 deoxy-D-[1-14C]glucose method. Linopirdine administration in naive rats (0.01, 0.1, or 1.0 mg/kg, s.c.) did not significantly alter cerebral glucose metabolism in any of the regions analyzed. Since linopirdine protects against hypoxia induced passive avoidance deficits in rats, we also examined the effects of linopirdine on cerebral metabolism after the rats were exposed to 30 min of hypoxia. Glucose metabolism was not significantly altered after hypoxic exposure, except for a small increase in some brain regions. Linopirdine administered after hypoxia decreased glucose metabolism in the hippocampus, limbic cortex, ventral hippocampal commissure, medial septum, striatum, subthalamic nucleus, zona incerta, lateral habenula, cerebral cortex, cerebellar vermis and a few thalamic nuclei. Statistically significant effects of linopirdine on glucose metabolism were observed in 22 of 56 brain regions sampled. In hypoxia-exposed rats, linopirdine altered glucose metabolism in brain regions that are implicated in learning and memory and are affected in Alzheimer's disease. Several of the affected regions are associated with the cholinergic system and may play a role in the cognitive enhancing properties of linopirdine. PMID- 8402202 TI - Modulation of ethanol neurotoxicity by nerve growth factor. AB - Dorsal root ganglion (DRG) neurons were cultured with varying concentrations of ethanol and NGF. At low concentrations of NGF (0.1 ng/ml) moderate initial ethanol levels (250 mg/dl) significantly suppressed neurite outgrowth. Higher NGF concentrations (5 ng/ml) protected against this neurotoxicity. At this higher NGF concentration, neuronal survival was not significantly affected by exposure to 0.25-4 g/dl ethanol, although survival was significantly diminished at 5 and 6 g/dl. Neurite outgrowth was a more sensitive indicator of ethanol neurotoxicity in this population, with significant decreases in process extension seen with 1 g/dl ethanol. When cultures were supplemented with 10 ng/ml NGF, however, process elaboration was significantly greater at 1 g/dl ethanol than that measured with 5 ng/ml NGF, and in fact did not differ from NGF controls. These studies indicate that NGF can provide neuroprotective effects against ethanol toxicity under these conditions. The results are discussed in relation to other recent reports of trophic factor neuroprotection. PMID- 8402203 TI - Time-optimized analysis of slit-scan chromosome profiles on a general-purpose personal computer. AB - Slit-scan flow cytometry provides a method to analyze large numbers of metaphase chromosomes in a relatively short time due to morphological features. The high detection rate requires fast computing for on-line analysis. Up to now, this has been achieved using special-purpose computers, parallel systems or other complex hardware. Here, we describe an algorithm that can be implemented on a general purpose personal computer. Digitized chromosome profiles can be classified by several criteria especially for the detection of chromosome abnormalities in biological dosimetry. A data set of approximately 4600 profiles was used. Programming in assembler results in an average computing time of about 600 microseconds per profile. Thus on-line evaluation of slit-scanning data appears to become feasible for many flow cytometers running nowadays. PMID- 8402204 TI - Locating well-conserved regions within a pairwise alignment. AB - Within a single alignment of two DNA sequences or two protein sequences, some regions may be much better conserved than others. Such strong conservation may reveal a region that possesses an important function. When alignments are so long that it is infeasible, or at least undesirable, to inspect them in complete detail, it is helpful to have an automatic process that computes information about the varying degree of conservation along the alignment and displays the information in a graphical representation that is readily assimilated. This paper presents methods for computing several such 'robustness measures' at each position of a given alignment. These methods are all very space-efficient; they use only space proportional to the sum of the two sequence lengths. To illustrate their effectiveness, one of the methods is used to locate particularly well conserved regions in the beta-globin gene locus control region and in the 5' flank of the gamma-globin gene. PMID- 8402205 TI - Multiple alignment of sequences on parallel computers. AB - A software package that allows one to carry out multiple alignment of protein and nucleic acid sequences of almost unlimited length and number of sequences is developed on C-DAC parallel computer--a transputer-based machine. The farming approach is used for data parallelization. The speed gains are almost linear when the number of transputers is increased from 4 to 64. The software is used to carry out multiple alignment of 100 sequences each of alpha-chain and beta-chain of hemoglobin and 83 cytochrome c sequences. The signature sequence of cytochrome c was found to be PGTKMXF. The single parameter, multiple alignment score, S, has been used to categorize proteins in different subfamilies and groups. PMID- 8402206 TI - A numerical method for allocating microbial isolates to strain types when characterized by typing methods that are not 100% reproducible. AB - Many methods for typing microbial strains are not 100% reproducible. This can create problems when deciding whether different groups of isolates are really distinct or represent typing errors or variation of a single strain. Neither hierarchical clustering nor iterative partitioning methods are suited for analysing such data. A novel iterative partitioning method is described which allows for the uncertainty of the typing method in use. Before grouping strains, the maximum dimension of the groups is set based on a previous knowledge of the typing method's reproducibility. Isolates are only allocated to a group if they differ from that group's typical strain type by less than the number of reaction differences required to distinguish between two strains. In a series of Monte Carlo studies the accuracy of strain allocation was found to be very good, even when the two groups were situated close to each other. PMID- 8402207 TI - Discovering simple DNA sequences by the algorithmic significance method. AB - A new method, 'algorithmic significance', is proposed as a tool for discovery of patterns in DNA sequences. The main idea is that patterns can be discovered by finding ways to encode the observed data concisely. In this sense, the method can be viewed as a formal version of the Occam's Razor principle. In this paper the method is applied to discover significantly simple DNA sequences. We define DNA sequences to be simple if they contain repeated occurrences of certain 'words' and thus can be encoded in a small number of bits. Such definition includes minisatellites and microsatellites. A standard dynamic programming algorithm for data compression is applied to compute the minimal encoding lengths of sequences in linear time. An electronic mail server for identification of simple sequences based on the proposed method has been installed at the Internet address pythia/anl.gov. PMID- 8402208 TI - SCUM--simulation of cyanobacterial underwater movement. AB - This paper describes the initial development of a computer simulation model of the vertical and lateral movement of a cyanobacterial bloom. Cyanobacteria actively regulate their position vertically within a water column by changing their buoyancy state in response to changing photosynthetic rates. Additionally the cyanobacteria are liable to lateral movement due to wind-induced currents and turbulence in the surface layers. The model may be applied to a range of water bodies under different wind environments. Initial results predict that periods of severe lake mixing encourages overbuoyancy in Microcystis, resulting in the rapid formation of surface scums. Oscillatoria respond slower to changes in near surface mixing and are liable to become entrained quickly within weak and turbulent currents. The model results agree well with published field studies. PMID- 8402209 TI - Doing sequence analysis with your printer. AB - The software package RSVP (Rapid Sequence Visualization in PostScript) has a suite of visually oriented sequence analysis routines implemented entirely in the page description language PostScript, a widely used standard that is built into many printers. RSVP is thus a relatively platform-independent tool for providing a 'quick look' at sequence data, using form and color to help point out patterns, in advance of more sophisticated sequence analyses. PMID- 8402210 TI - CURVATURE: software for the analysis of curved DNA. AB - Software is presented to plot the sequence-dependent spatial trajectory of the DNA double helix and/or distribution of curvature along the DNA molecule. The nearest-neighbor wedge model is implemented to calculate overall DNA path using local helix parameters: helix twist angle, wedge (deflection) angle and direction (of deflection) angle. The procedures described proved to be very convenient as tools for investigation of a relationship between overall DNA curvature and its gel electrophoretic mobility. All parameters of the model had been estimated from experimental data. Using these wedge parameters the program takes, as input, any DNA sequence and calculates the likely degree of curvature at each point along the molecule. This information is displayed both graphically and in the form of simplified representations of curved double helices. The Software, CURVATURE, can thus be used to investigate possible roles of curvature in modulation of gene expression and for location of curved portions of DNA, which may play an important role in sequence-specific protein--DNA interactions. PMID- 8402211 TI - SCAMP: a general-purpose simulator and metabolic control analysis program. AB - SCAMP is a general-purpose simulator of metabolic and chemical networks. The program is written in C and is portable to all computer systems that support an ANSI C compiler. SCAMP accepts metabolic models described in a biochemical language, and this enables novice as well as experienced users rapidly to build and simulate metabolic systems. The language is sufficiently flexible to enable other types of model to be built, e.g. chemostat or ecological models. The language offers many facilities, including: the ability to describe metabolic pathways of any structure and possessing any kinetics using normal chemical notation; optionally build models directly from the differential equations; differing compartment volumes; access to flux, concentration and rate of change information; detection of conserved cycles; access to all coefficients and elasticities of metabolic control analysis; user-defined forcing functions at the model boundaries; user-defined monitoring functions; user-configurable output of any quantity. From the model description SCAMP can either generate C code for later compilation to produce fast executable stand-alone models or run-time code for input to a run-time interpreter for immediate execution. The simulator also incorporates an inbuilt symbolic differentiator for evaluating the Jacobian and elasticity matrices. PMID- 8402212 TI - GeneView: multi-language human gene mapping library with a graphical user interface. AB - GeneView is a newly developed human gene mapping library system that works on an X-Window platform. This system is designed for researchers who routinely utilize gene mapping data in the laboratory but are unfamiliar with computer technology. GeneView offers various features, including friendly user interface, fast operation and visualization facility. Genetic loci are displayed graphically with an idiogram. This system can be operated in multiple languages. A current version supports Japanese and English, and can be easily expanded to include other languages. A prototype of GeneView is now installed on a SPARC (Sun4) workstation. Because of the X-Window platform, GeneView should be compatible with various types of workstations. PMID- 8402213 TI - A computer program to calculate and design oligonucleotide primers from amino acid sequences. AB - A computer program to generate oligonucleotide sequences from peptide data is presented. The program, PrimerGen, reverse-translates a peptide string into its corresponding degenerate deoxyribonucleotide counterpart, and generates sequences of oligonucleotide molecules which can be synthesized and either used as probes for hybridizations or as primers in polymerase chain reactions. PMID- 8402214 TI - 3DVISION: a program for display of three-dimensional wire models on a PC with VGA display. PMID- 8402215 TI - A simple method for accessing and transferring nucleotide database files. PMID- 8402216 TI - A fast graphics printing program for neurophysiological data. AB - A program was written in assembly language for fast graphics screen dump of neurophysiological data during and after experiments. This program takes approximately 3 s to plot a 1024 x 768 graphics image. A resident program has also been produced, which allows a screen graphics image to be printed by using a keyboard command. A Pascal-compatible object file is available to interface the program with Turbo-Pascal programs. PMID- 8402217 TI - A computer program in BASIC for estimation of LD50 of a chemical by Dixon's up and down method. PMID- 8402218 TI - OLIGOGET--a computerized database system for controlling oligonucleotide production. PMID- 8402219 TI - Chemotherapy with high-dose cytosine arabinoside and mitoxantrone for poor prognosis myeloid leukemias. AB - Forty-seven patients with poor-prognosis myeloid leukemias received induction therapy with high-dose cytosine arabinoside (HDara-C), 1.5-3.0g/m2 for 8-10 doses, and mitoxantrone (DHAD), 12-15 mg/m2 for 3 doses. Complete remissions were achieved in 21 [45%, 95% confidence interval (CI) 30.2-59.9%] of the patients, including 11 of 14 with acute myelogenous leukemia (AML) in first relapse (79%, 95% CI 49.2-95.3%), 4 of 8 with refractory anemia with excess blasts in transformation (RAEBiT) (50%, 95% CI 15.4-84.6%), and 4 of 6 (67%, 95% CI 22.3 95.7%) previously untreated elderly AML patients. Patients with secondary AML and advanced chronic myelogenous leukemia had a very low response rate. The incidence of reversible toxicity was low and only 3 treatment-related deaths occurred. After reinduction, 8 of 9 AML patients < or = 60 years of age were ultimately able to undergo intensive therapy and either autologous 4 hydroperoxycyclophosphamide-purged bone marrow (7 patients) or peripheral blood stem cell (1 patient) transplantation with satisfactory hematological recovery. We conclude that HDara-C and DHAD is an effective antileukemic regimen in selected AML and RAEBiT patients, and that its use may allow subsequent successful autologous BMT in appropriate patients. PMID- 8402220 TI - Chemiluminescence production by neutrophils and immune complex levels in cancer patients. AB - We studied the production of chemiluminescence (CL) by peripheral blood neutrophils from 24 normal subjects, 13 lung cancer patients with clinical stages (CS) III and IV, and 27 breast cancer patients with CS II, III, and IV. Evaluations were made before chemo- and radiotherapy treatments. CL was expressed as: baseline of the record background activity; area under the curve (AUC) of opsonized zymosan response; maximum peak (MP) response; time to MP (TMP); and total time (TT). Simultaneously, circulating immune complexes (CIC) in sera were evaluated by polyethylene glycol precipitation technique. The results demonstrated that lung cancer patients with CS III had an increase of TMP and TT values, while breast cancer patients with CS IV showed the lowest values of AUC and MP compared with the control group. The CIC levels were increased in all the cancer patients. There was no correlation between the baseline CL activity and the levels of immune complexes. PMID- 8402221 TI - Guanosine potentiates the antiproliferative effect of cytosine-beta-D arabinofuranoside in melanoma cell lines. AB - Guanosine is shown to potentiate markedly the antiproliferative effect of cytosine-beta-D-arabinoside (ara-C) on B16 F10 mouse and SKMEL-28 human melanoma cell lines. Several metabolic consequences of the synergistic interaction between ara-C and guanosine on cell growth were determined in B16 F10 mouse melanoma cells. Treatment of the cells with guanosine for 24 hr resulted in an increase in the percentage of cells in the S phase of the cell cycle, a threefold increase in intracellular GTP concentration, and an increase in the incorporation of ara-C into acid-insoluble material and phosphorylated metabolites. These findings suggest that guanosine potentiates the growth-inhibitory effect of ara-C in B16 F10 melanoma cells by increasing the intracellular concentration of its active metabolites. PMID- 8402222 TI - Effects of 5-fluorouracil and a combination of tegafur and uracil (UFT) on nucleotide metabolism in L1210 ascites tumor. AB - Changes in the deoxyribonucleotide pools following a single oral administration of 13 mg/kg of 5-fluorouracil (5-FU) or of 64.8 mg/kg of UFT (a mixed compound of tegafur and uracil) were investigated. We compared their pharmacodynamics and effects on nucleotide metabolism in L1210 ascites tumor on day 3 after intraperitoneal tumor inoculation. The intracellular dTTP pool decreased to half the control level 1-6 hr after the administration of 5-FU. The dTTP pools rapidly recovered after 6 hr. In contrast, UFT kept the intracellular dTTP level to 1/3 to 1/2 of the control level for 24 hr. Either drug elevated the intracellular dATP pools, but decreased dCTP pools. UFT influenced the intracellular dATP and dCTP levels longer than 5-FU. Orally administered UFT seemed to exert a longer and more potent inhibitory effect on thymidylate synthetase than equimolar 5-FU. In view of these results, we suggest that UFT could be a more potent chemotherapeutic drug than 5-FU in oral administration. PMID- 8402223 TI - Treatment of childhood acute nonlymphoblastic leukemia: a review. AB - Recommendations for the treatment of ANLL in general, and in pediatrics more specifically, are still conflicting. Overall, a child diagnosed with ANLL has a 35-50% of remaining alive long term, without disease. Induction chemotherapy with cytarabine and daunorubicin will achieve remission in about 80% of children with ANLL. So far, intensifying induction chemotherapy by adding more agents has not changed this result significantly. Major changes in induction chemotherapy may come from either new chemotherapeutic agents or biological agents that hasten bone marrow recovery after treatment. Such an approach might allow more dose intensive drug administration without increased toxicity. Another question that is slowly being answered with the ongoing trials is the one concerning maintenance. So far, all the pediatric trials that have tried to shorten the maintenance therapy of this disease were able to do so without jeopardizing the final outcome. The final optimal minimal duration of therapy has yet to be established. The best therapy for a patient who has achieved a remission is still the most difficult question regarding the treatment of this disease. So far, allogeneic BMT has yielded the best results, with a disease-free interval varying between 55 and 65%. However, one-third of these patients have chronic GvHD and, therefore, a somewhat diminished quality of life. As preparative regimens and marrow purging protocols evolve, the results of autologous bone marrow transplantation seem to be improving, with disease-free intervals of 35-50% reported. PMID- 8402224 TI - Anaplastic Ki-1 + large-cell lymphoma. PMID- 8402225 TI - Ostomy patient management: care that engenders adaptation. AB - The patient requiring ostomy surgery has needs above and beyond the typical perioperative needs of an individual experiencing major abdominal surgery. These needs may be categorized as technical ostomy management, information, and emotional support. The health care team managing ostomy patients must be prepared to address the patients' need in these three categories. This can be accomplished when the multidisciplinary team establishes a comprehensive plan of care that consists of five elements: information, technical expertise, psychosocial support, referrals, and surveillance. Each element plays a critical role in the patients' adaptation to ostomy surgery. Pertinent aspects of each element are described. By incorporating these elements into the plan of care, the health care team engenders patient adaptation to ostomy surgery. PMID- 8402226 TI - Nonclassical platinum antitumor agents: perspectives for design and development of new drugs complementary to cisplatin. AB - Studies over the last few years have shown that the range of platinum complexes with useful cytotoxicity and antitumor activity is not strictly limited to structural analogs of cisplatin. In general, we can expect that cells will process structurally different species in different manners. The metabolic chemistry and DNA binding will be altered in in comparison to the cis-[PtX2 (amine)2] class. This point is of particular importance because any altered pattern of antitumor activity of structural analogs of cisplatin is likely to be due to unpredictable pharmacokinetic, rather than truly mechanistic, factors. The fact that discrete cisplatin-DNA adducts vary in their biological activity further supports the hypothesis that complexes structurally dissimilar to cisplatin may produce biological activity complementary to the parent drugs. The mechanism of action of nonclassical complexes is different from that of cisplatin and its analogs. Their pattern of antitumor activity is also altered with respect to cisplatin--thus, not all platinum-containing drugs need necessarily be similar in their clinical profile to cisplatin. Note that both the dinuclear bis(platinum) complexes and the trans complexes give their own distinct patterns of tumor specificity--different from cisplatin and each other (see Tables 1 and 3). New cytotoxic mechanisms for platinum complexes may also be placed in context with cisplatin resistance. Modes of overcoming cisplatin resistance may reside at the various levels of uptake, interaction with "detoxifying" intracellular thiols, and DNA repair. Likewise complexes with novel mechanisms of action may circumvent resistance by more than one unique route. Indeed, the three major routes to resistance are all affected to varying degrees by the complexes outlined above. From the discovery of cisplatin, the development of analogs has essentially been an empirical exercise. Because of their similar mechanism of action, much comparison has been made between platinum complexes and the classic alkylating agents. Yet the alkylating agents represent a good example where a number of structurally distinct drugs with different anticancer activities are clinically available. This desirable feature may be achieved for platinum complexes by emphasis on complexes structurally dissimilar to the presently used agents. The dinuclear bis(platinum) complexes and mononuclear complexes in the trans geometry are of special interest. Comparison of common features and differences between different classes may point to guidelines for the rational design of complexes with a different spectrum of clinical antitumor activity to cisplatin and activity against cisplatin-resistant tumors. PMID- 8402227 TI - Retinoids and cancer: a basis for differentiation therapy. PMID- 8402228 TI - Interleukin-1: therapeutic potential for solid tumors. PMID- 8402229 TI - Interleukin-3: structure and function. PMID- 8402230 TI - The biology of interleukin-5 and its receptor. AB - IL-5 is primarily a T-cell-derived cytokine that has multiple regulatory functions on eosinophils and (in the mouse) on antibody-secreting B cells. A complex network of cytokines appear to control transcription of the gene for IL-5 and its production. Abnormally high levels of this cytokine are associated with infections with tissue-dwelling parasites and a diverse group of hypereosinophilic conditions of no known etiology. Our understanding of the biological role of IL-5 in the regulation of Ig production and the development of immunity to parasites is far from complete, but basic knowledge of its action at the cellular level is accumulating and will be critical for the intelligent application of immunotherapy with IL-5 or antibodies to IL-5 in infectious, neoplastic, and possibly other diseases. PMID- 8402231 TI - Interleukin-9: a T-cell growth factor with a potential oncogenic activity. PMID- 8402232 TI - Ultrasonography of the ankle and the hindfoot. AB - High-resolution ultrasonography (US) has proved an excellent noninvasive and inexpensive modality for examining the extremities. This pictorial essay illustrates the US appearance of bony and soft-tissue abnormalities of the ankle and the hindfoot. The role of US as an adjunct technique to complement conventional radiography is stressed. The advantages of US in examining traumatic, inflammatory and infectious lesions and soft-tissue masses are discussed. PMID- 8402233 TI - Imaging of the salivary glands. AB - The most common conditions affecting the salivary glands are calculi, inflammation and tumours. Plain radiography may be useful, especially in showing calculi. Sialography is the examination of choice for inflammatory diseases, whereas ultrasonography, computed tomography and magnetic resonance imaging are particularly useful in evaluating masses in the salivary glands. Radionuclide studies are sometimes helpful in evaluating inflammatory or tumoral diseases. Each technique has its place, and the methods are often complementary. In this review article the authors present the techniques that they use in evaluating diseases of the salivary glands. PMID- 8402234 TI - Ultrasonographically guided biopsy of small intra-abdominal masses. AB - Ultrasonography (US) has traditionally been considered inferior to computed tomography (CT) for guidance in fine-needle aspiration biopsy (FNAB) of small intra-abdominal masses. To assess the validity of this view, the results obtained for 58 consecutive patients referred for US-guided FNAB of intra-abdominal masses of diameter 25 mm or less were retrospectively analysed. In each case two to four (usually three) aspirations were initially performed with a 22-gauge needle; sufficient tissue for diagnosis was obtained in 53 of the 58 cases (91%). The procedure had to be repeated in the other five patients to acquire diagnostic tissue. Apart from some minor abdominal pain, no complications were encountered. Proof of the cytologic diagnosis was available for 49 of the 58 patients (84%)- from histologic examination in 22 cases and from the appropriate clinical and radiologic course in 27. A sensitivity of 95% and a specificity of 90% for the technique were found for the patients for whom adequate follow-up information was available. The authors conclude that FNAB of intra-abdominal masses measuring 25 mm or less can be accurately and safely performed under US guidance. PMID- 8402235 TI - Dynamic abdominal computed tomography: "top-down" compared with "bottom-up" imaging. AB - Dynamic contrast-enhanced computed tomography (CT) of the liver is usually performed from the top down, and there is usually less opacification of the top slices than the more inferior slices. The authors reasoned that reversing the direction of scanning would allow more time for the parenchyma at the top of the liver to become enhanced and would result in better opacification of the parenchyma and the hepatic veins. To test this hypothesis they assigned 32 patients to either a "top-down" or a "bottom-up" group; each patient then underwent unenhanced and contrast-enhanced CT. The method of intravenous administration of contrast agent was identical in the two groups and consisted of rapid injection of 135 mL of iohexol by a power injector; scanning was initiated 30 seconds after the injection was started. Attenuation measurements were obtained at specified areas in the liver and in the hepatic and portal veins. The degree of enhancement was calculated at each site and was compared between groups. The upper portion of the liver showed significantly greater enhancement in the bottom-up sequence than in the top-down sequence (123% and 22% respectively; p < 0.01); the entire liver showed excellent enhancement in the bottom-up sequence. In addition, the hepatic veins showed significantly greater conspicuity in the bottom-up sequence (p < 0.01). The authors conclude that the dynamic bottom-up method yields excellent hepatic enhancement and vascular opacification and is therefore superior to the standard top-down method. PMID- 8402236 TI - Computed tomographic and cytologic assessment of cystic pancreatic neoplasms: a difficult preoperative diagnosis. AB - The computed tomography (CT) findings or cytologic results, or both, for 21 patients with cystic pancreatic neoplasm (4 with microcystic cystadenoma, 6 with macrocystic mucinous cystadenoma, 10 with macrocystic mucinous cystadenocarcinoma and 1 with a papillary cystic neoplasm) were reviewed. CT scans for 14 of the patients were reviewed by two gastrointestinal radiologists who were blinded with respect to the patients' identities and the diagnoses. The radiologists used previously published criteria for distinguishing between microcystic and macrocystic neoplasms. Of the four cases of microcystic cystadenoma, two were correctly diagnosed by one radiologist, and one was correctly diagnosed by the other. Three and four cases respectively of five cases of macrocystic cystadenoma were correctly identified, as were three and five of five cases of macrocystic cystadenocarcinoma. Cytologic evaluation of samples from fine-needle aspiration biopsy had been performed for 15 of the patients, and these records were reviewed. One of three cases of microcystic cystadenoma, two of four cases of macrocystic cystadenoma, five of seven cases of macrocystic cystadenocarcinoma and the papillary cystic neoplasm were correctly diagnosed on the basis of the cytologic findings. The combination of CT and cytologic assessment is helpful in distinguishing different types of cystic pancreatic neoplasms, but there is significant overlap among the clinical and radiographic features of these lesions, and therefore operative assessment is often necessary. PMID- 8402237 TI - Demonstration of renal tubular flow by selective angiographic computed tomography. AB - A high-speed computed tomography scanner was used to study the bulk flow of a small bolus of nonionic contrast medium through the renal tubules of five dogs. A 0.5-mL bolus of iohexol 300 (150 mg iodine) was injected rapidly into the renal artery, and transverse images of the kidney were obtained at 15-second intervals over 300 seconds. The mean attenuation values measured in regions of interest in the papilla and the cortex were displayed as a function of time. Curves from the papillary regions of interest showed two peaks, the first corresponding to the passage of the bolus through the loops of Henle that extend into the papilla and the second to the passage of the bolus through the terminal collecting tubules. The cortical regions of interest showed the cortical return peak, which corresponded to the passage of the bolus through the distal convoluted tubules. The peaks generated by this method can be used to measure the transit times of the bolus from the glomeruli to the loops of Henle in the papilla, from the loops of Henle to the distal convoluted tubules, and from the distal convoluted tubules to the distal collecting tubules. The mean total transit time in dehydrated dogs was 169 seconds. The method is compared with contrast-enhanced magnetic resonance imaging (with gadolinium diethylenetriaminepentaacetic acid), which has been used by other researchers for the same purpose. PMID- 8402238 TI - Comparison of real-time ultrasonography and coronal computed tomography in the diagnosis of orbital fractures. AB - Real-time ultrasonography (US) was used to evaluate the bony orbit in 19 patients who had sustained orbital trauma. The ability of US to demonstrate clinically significant orbital fractures was compared with that of thin-section coronal computed tomography (CT); the latter method was treated as definitive. US had a sensitivity of 92%, a specificity of 100% and a positive predictive value of 100%. Quantification of the size of fragments by the two methods yielded similar results. Real-time US adequately displayed the anatomic features of the orbit and detected clinically significant fractures. This technique may have a role in post traumatic imaging of the orbit when coronal CT is not possible. PMID- 8402239 TI - Imaging of spinal cord hemangioblastomas. AB - Hemangioblastomas, which are associated with von Hippel-Lindau disease, represent the third most common type of intramedullary spinal cord tumour. The authors reviewed the radiologic findings in five cases of surgically proven spinal cord hemangioblastoma. Magnetic resonance imaging (MRI) could be used to differentiate the various components of these tumors, including the tumour nodule, edema, cysts and syringomyelia. The tumour nodule could be identified as an isointense area surrounded by edema, a cyst or syringomyelia, or as an area adjacent to a signal void (which represented feeding and draining vessels). Tumour nodules demonstrated intense enhancement when gadolinium diethylenetriaminepentaacetic acid was administered. Edema appeared as an ill-defined region of moderately low signal intensity in T1-weighted images and moderately high signal intensity in T2 weighted images. Cysts and syringomyelia appeared as well-defined regions of low signal intensity in T1-weighted images and high signal intensity in T2-weighted images. MRI allows the radiologist to assist the surgeon in determining preoperatively the level of laminectomy appropriate to resect the tumour. Preoperative embolization of the posterior spinal arteries, facilitated by angiography, can be important in reducing perioperative hemorrhage. PMID- 8402240 TI - Indonesian type of metaphyseal dysplasia. AB - The authors describe two Indonesian siblings affected by a hitherto undocumented form of metaphyseal dysplasia that simulates enchondromatosis. The notable clinical features were short stature and lateral bowing of the lower extremities. These features were readily recognizable at birth. The radiographic examinations showed that the metaphyseal anomalies were localized predominantly in the lower extremities, the hip joints being the most severely affected. The upper extremities showed only minimal abnormalities. The name "Indonesian type of metaphyseal dysplasia" is suggested for this autosomal recessive bone disorder. PMID- 8402241 TI - Asynchronous bilateral epidural hematomas. AB - The immediate and delayed sequelae of bilateral lateral fractures of the cranial vault were illustrated in a patient in whom asynchronous bilateral epidural hematomas developed after an automobile accident. The importance of examining computed tomography images obtained with both brain and bone window settings is stressed. PMID- 8402242 TI - Adenoid cystic carcinoma: an unusual sellar mass. AB - Adenoid cystic carcinoma represents approximately 20% of tumours of the minor salivary glands. It can originate in the sphenoidal sinus and invade the sella turcica. The authors describe a patient in whom a tumour of this type was shown by computed tomography. PMID- 8402243 TI - Importance of routine T2-weighted or T2*-weighted coronal images in magnetic resonance imaging of the knee. AB - Magnetic resonance imaging of the knee is most commonly performed with T1 weighting in the coronal plane. This type of imaging is insensitive to meniscocapsular separation and tears of the medial collateral ligament. T2 weighted and T2*-weighted coronal imaging is sensitive to these abnormalities and should be a part of the routine protocol for imaging the knee. PMID- 8402244 TI - Residents' corner. Answer to case of the month #21. Transient osteoporosis of the hip. PMID- 8402245 TI - Informed consent. PMID- 8402246 TI - The dizzy artifact. PMID- 8402247 TI - The psychological abuse of latency age children: a survey. AB - Most states have included sections on psychological abuse or emotional maltreatment in their child abuse statutes, however, interest in this type of abuse has lagged in litigation, treatment, or research. Lack of public sanctions to examine or prosecute these cases may be a reason for this reluctance. This research project aimed at identifying potential definitions of psychological abuse, by submitting vignettes with adult behaviors to be rated as abuse or not by a group of citizens and also comparing these results with a professional social work cohort. Both groups identified nine types of adult behavior to be abuse. There was no difference on the ratings between respondent groups (citizen social work; age groups, experience with child abuse, and parenthood), except for gender. Female respondents rated the vignettes to be child abuse, serious, and wanted more drastic intervention than male respondents. Support existed for intervention in these cases. PMID- 8402248 TI - Conflict behaviors of maltreated and nonmaltreated children. AB - Maltreated and nonmaltreated children (ages 3-7 years) were paired for brief play sessions involving a single desirable object. Facial, verbal, and physical actions used while negotiating access to the object were examined. Results showed that the maltreated-nonmaltreated pairs were able to engage in a negotiation process resulting in approximately equal sharing of the object. The differences found between maltreated and nonmaltreated children primarily suggested that maltreated children were somewhat hesitant to engage their partners. Results also showed that children's scores on a facial expression recognition task predicted greater responsiveness to their partners' indication of reluctance to relinquish the object. PMID- 8402249 TI - Psychiatric diagnoses of self-reported child abusers. AB - In order to evaluate lifetime psychiatric diagnoses and prevalence of dyssocial behaviors among self-reported child abusers, three large databases of clinical, community, and family study subjects were examined. Subjects who had acted as parents and who reported any episodes of child battery were compared to those without any history of child battery on prevalence of psychiatric disorders and dyssocial behaviors. Overall, 4% of subjects from the community sample reported child abuse. Abusers not selected through alcoholism treatment were more likely to receive diagnoses of alcoholism, antisocial personality disorder, and major depression. Those selected through alcoholism treatment were more likely to have antisocial personality disorder. Abusers in general were found to have a history of disciplinary problems, property destruction, and as adults to engage in other violent behaviors. It was concluded that self-identified child abusers have increased lifetime rates of antisocial personality disorder, alcoholism, and depression. The association between child abuse and other violence is not explained by selection of cases through the medical or legal systems. PMID- 8402250 TI - Recall of abuse in childhood and three measures of dissociation. AB - Recalled abuse was examined in relation to dissociation in a sample of 312 undergraduates. The Physical Abuse Scale of the Assessing Environments III (Berger & Knutson, 1984) and a newly developed sexual abuse questionnaire measured recalled physical and sexual abuse. The Harvard Group Scale of Hypnotic Susceptibility (Shor & Orne, 1962), Tellegen Absorption Scale (Tellegen, 1982), and Dissociative Experiences Scale (Bernstein & Putnam, 1986) measured dissociation. Significant correlations were found between the abuse variables and the dissociation variables, for all the measures except the Harvard Group Scale. Prevalence of sexual abuse, but not physical abuse was found to be significantly higher for females than males. PMID- 8402251 TI - Juvenile delinquents' perceptions of childhood parental rearing patterns. AB - This study compared individuals who have been identified as engaging in delinquent behavior, but who perceive their childhood differently. It examined the differentiating family and social characteristics between delinquents who were maltreated in childhood (based on self-report data) and delinquents who were not maltreated. The sample consisted of the entire population of juvenile delinquents (N = 287) in all institutions and prisons in Greece. Childhood abuse and neglect were defined on the basis of perceived parental rearing patterns as assessed by the EMBU questionnaire, and groups of abused/nonabused and neglected/non-neglected delinquents were formed. Perceived parental abuse and/or psychological neglect (paternal and/or maternal) were found to be associated with various family and social characteristics. Specific parameters connected to the Greek societal and cultural context have also been identified. PMID- 8402252 TI - Components of child and parent interviews in cases of alleged sexual abuse. AB - Considerable discussion in child sexual abuse evaluation centers around minimizing the number of victim interviews, however, the completeness of one professional's interview is not always addressed. In this study, professionals (medical, social, law enforcement/legal, mental health) indicated what components they included in their interviews of sexual abuse victims and their parents. Components included: details of abuse, family relationships, school situation/performance, child's development (toilet training, bed wetting, language development, etc.), child's knowledge of body part names, child's knowledge of body part functions, child-rearing practices (bathing, sleeping), medical history (illnesses, surgery), behavior problems, psychological symptoms (nightmares, sadness), physical complaints (pain, discharge), parental teaching about sex, and child's access to sexually explicit television or magazines. Interview components varied by profession, consistent with expected professional bias. This study demonstrates some professional bias in interviewing children and parents and suggests that having a single interview or interviewer may not always be optimal for a thorough evaluation. Increased communication and teamwork among professionals, and cross-training among disciplines could facilitate both the recognition of appropriateness of some "multiple interviews" as well as provide efforts to consolidate interviews when possible. PMID- 8402253 TI - Adoption at birth: prevention against abandonment or neonaticide. AB - A study was carried out between 1987 and 1989 to try to understand why women choose to give their infants up for adoption at birth and the psychodynamics of this decision. The interviews of 22 female subjects were based on psychoanalytic methodology. They revealed that the motives behind such a choice arose from a specific set of symptoms: The denial of pregnancy and fantasies of violence towards the fetus. These symptoms were the result of psychological and sexual traumas that the subjects experienced during their childhood in negligent or incestuous families. Eighteen of the subjects were able to take advantage of the French law permitting anonymous and cost-free delivery, then the adoption of the baby. However, four of the subjects denied their pregnancies so efficiently that childbirth took them by surprise. The sudden discovery of the reality of the newborn led them to neonaticide. This study clarifies the confusion that exists between the abandonment of a child in a public place, and the choice of adoption at birth, the aim of which is to protect the infant from the risk of violence or future neglect. PMID- 8402254 TI - "Normal" childhood sexual play and games: differentiating play from abuse. AB - Recent recognition of child-to-child and adolescent-to-child sexual abuse raises the question, for the courts, educators, clinicians, and lay individuals, where do we draw the line between normal childhood sexual play, and abuse. This paper presents the results of a survey on normative childhood sexual play and games experiences that was distributed to 300 undergraduates at an all women's college. One hundred-twenty-eight returned the survey, 85% of whom described a childhood sexual game experience. Of these women, 44% described cross-gender play and there was a trend for women who had described cross-gender experiences to have seen the play as involving persuasion, manipulation, or coercion. A strong relationship was found between abuse and cross-gender play. Level of physical involvement in the game was correlated with perceptions of normality. A typology of six kinds of sexual play experiences was derived. Results are discussed in terms of their relation to differentiating childhood sexual abuse from play and gender socialization influences relating to the role rehearsal of coercive or manipulative relationships. PMID- 8402255 TI - Parental involvement in sexual abuse prevention education. AB - Parental knowledge of child sexual abuse and interest in educating themselves and their children for primary prevention was studied. Fifty-one mothers and 50 fathers of preschool and day-care center children were interviewed. Parents wanted to be the primary educators of their child, but demonstrated a lack of knowledge about important sexual abuse issues and planned to discuss only the least threatening topics. Mothers and fathers obtained most of their information about sexual abuse from the media. However, parents were open to numerous methods and referral sources for prevention education. One dependent variable, sex of parent, showed significant differences on sources of sexual abuse information; preferred educators for their children; number and types of topics acceptable for children to learn about at preschool ages or other times; the number of topics parents planned to discuss; sources parents would respond to most for a parent program; and factors in parent's decision to attend parent programs. There were no significant differences based upon the sex of the child and no significant interaction effects between sex of parent and sex of child. Further research on parent motivation for their own education and the education of their children is indicated. PMID- 8402256 TI - Child sexual abuse and gender differences: attitudes and prevalence. AB - A great deal of attention has been focused on child sexual abuse in recent years. The present study reports attitudes and beliefs of 824 male and female college students concerning sexual abuse, and the participants' own experience with such abuse. Results indicate that women have much stronger pro-social beliefs, attitudes, and emotional reactions to sexual abuse and to abusers and victims than men do. In addition, 6% of men and 13% of women participants reported that they had been sexually abused as children or adolescents, while equal proportions of males and females reported that they were sexually approached by a potential perpetrator. These results are discussed in terms of socialization experiences of males and females in our society, and the literature regarding helping behaviors and compassion for others perceived as helpless and in need of aid. In particular, females may be abused more often than males, even though both sexes are approached in equal frequency, because females have been socialized to be more compliant and responsive to the needs of others, while males are perpetrators more often than females because males have been socialized to be more aggressive, powerful, and dominant. PMID- 8402257 TI - Assessment of professionals' and nonprofessionals' attitudes toward child abuse in Croatia. PMID- 8402258 TI - Psychological maltreatment in the context of separation and divorce. AB - Children of separating or divorcing parents seldom escape suffering psychological stress, particularly when the parents are in open conflict. How much should such children be allowed to suffer? The author discusses the problem of defining the terms "emotional child abuse" and "psychological maltreatment" as they are used in the literature. Reviewing the common behavioral patterns that children in this situation resort to and the roles that they tend to take upon themselves, he attempts to distinguish between those instances of stress that can be regarded as acceptable and those that must be considered harmful. With respect to children in this situation, the following behavior patterns are discussed: (a) the inclination of the child to sacrifice itself for the sake of the parents, in particular for the "weaker" one; (b) the tendency of the children to reach agreements among themselves about how they should be divided up; (c) the phenomenon of parentization in which the child assumes the role of the substitute partner for one or the other parent; and (d) the child's discovery of both its power and its helplessness with respect to the situation and the feelings of guilt awakened by this discovery. PMID- 8402259 TI - [Change of the thermostability of renin by glycosylation]. AB - Inbred strains of mice, which produce high levels of submaxillary gland (SMG) renin have two renin genes, Ren 1 and Ren 2, per haploid genome, while strains with low levels of SMG renin have only the Ren 1 gene. Ren 1 codes for a glycosylated protein and Ren 2 codes for a highly homologous but unglycosylated and less thermostable protein. In order to determine if this difference in thermostability is related to the absence of glycosylation of renin-2 and/or to some amino acid difference between both renins we have compared the thermostability of renin-2 and a renin-2 mutant containing two potential N glycosylation sites added by in vitro mutagenesis. Both mutant and wild type renins were expressed in AtT20 cells. Wild type renin was significantly less stable than the glycosylated mutant form upon heating. Moreover, the kinetics of heat inactivation of the mutant, in vitro deglycosylated form was similar to that of the wild type renin. This result indicates that glycosylation affects the thermostability of renin. PMID- 8402260 TI - Expression of acetylcholinesterase molecular forms during in vitro differentiation of rabbit embryonic muscle cells. AB - We investigated the myogenic properties of rabbit myoblasts in culture, and examined particularly the expression of acetylcholinesterase (AChE). In the presence of horse serum, myoblasts proliferate quickly and begin to fuse by day 3 of culture. Differentiated myotubes presenting some spontaneous contractions are clearly visible from days 6 to 9. The cultures degenerate from day 10 onwards. The specific activity of AChE is low during the proliferation phase of myogenic cells, increases sharply as myotubes differentiated, then decreases as cultures degenerate. In 7-day-old cultures, globular forms are largely predominant and asymmetric forms, detected as A12, represent only 1% of the total activity. PMID- 8402261 TI - Lineage control of an integration site-dependent transgene combined with the beta globin locus control region. AB - In mice, the expression of integration site-dependent transgenes is controlled by host DNA elements. These may involve locus control regions (LCR), enhancers and silencers. We analysed the interaction between the LCR of the beta-globin locus (beta-LCR) and an integration site-dependent LacZ transgene (HPRTnlsLacZ). We obtained LacZ expression in erythroid cells of all transgenic mice, independent of temporal control, correlated to transgene copy-number. These results demonstrate that an integration site-dependent transgene can be region-controlled by the beta-LCR and establish that the beta-LacZ transgenic lines constitute a valuable source of genetically marked cells for the three erythroid series. This also extends to in vivo regulation in all three erythroid series the notion that the beta-LCR contains the information for region-control of non-erythroid genes. In non-erythroid lineages of all mice, we observed integration site-dependent LacZ expression. It suggests that a single LCR is not sufficient to insulate the LacZ gene from endogenous cis-acting control elements. PMID- 8402262 TI - Two yeast chromosomes are related by a fossil duplication of their centromeric regions. AB - A 15 kbp fragment of the Saccharomyces cerevisiae genome was cloned and localised to the centromeric region of chromosome XIV by genetic linkage and DNA sequencing. It had a strong sequence similarity and a conserved gene linkage and transcriptional orientation relatively to the centromeric region of chromosome III, indicating a fossil interchromosomal duplication of several linked genes. On chromosome XIV, the duplicated fragment included the centromere, four genes (FUN34, CIT1 and two tDNAs), one open reading frame (DOM34) and a truncated delta element. Additional inserts bearing unique genes were present on the centromeric region of chromosome III. The level of silent substitutions indicated a relatively ancient genetic separation, pre-dating the emergence of S. cerevisiae and S. douglasii as distinct species. The ensuing evolution of the duplicated regions retained strict sequence identity for the two tDNAs pairs, but was partially divergent for CIT1 and FUN34, and generated a probable pseudogenic equivalent of DOM34 on chromosome III. Extant multigenic duplications of this type might play an important role in the evolution of eukaryotic genomes. PMID- 8402263 TI - Familial case of 46,XX male and 46,XX true hermaphrodite associated with a paternal-derived SRY-bearing X chromosome. AB - The human testis-determining gene was recently isolated from a 35 kb region on the human Y chromosome which was present in four sex-reversed individuals, three XX males and one true hermaphrodite. One of the XX males and the true hermaphrodite were sibs. A more detailed molecular analysis of these two patients and their family for Y-DNA sequences including the testis-determining gene, SRY was performed. The father was found to harbor two copies of SRY, one on his Y chromosome and the other on his X chromosome located at Xp22 determined by in situ hybridization. Somatic cell hybrids were generated from peripheral blood lymphocytes. Analysis of Y chromosome-negative somatic cell hybrids from the XX male, the true hermaphrodite and their father, revealed that both the X and Y pseudo-autosomal boundaries were present. The present of both boundaries suggests than an unequal interchange of X and Y material occurred with the cross-over breakpoint located within the X pseudo-autosomal region. The paternal SRY-bearing X chromosome was transmitted to two of his children, a 46 XX true hermaphrodite and a 46,XX male. The presence of SRY on an X chromosome associated with two sex phenotypes strongly suggests that the phenotypic variability was caused by differential inactivation of the SRY-bearing X chromosome, thereby influencing SRY expression. PMID- 8402264 TI - The HNF1 C-terminal domain contributes to transcriptional activity and modulates nuclear localisation. AB - HNF1 is a homeoprotein that regulates the expression of a large number of liver specific genes. By performing transient expression assays with a series of C terminal deletion mutants and with a LexA-HNF1 fusion protein, we show that the C terminal half of HNF1 is necessary and sufficient for in vivo transcriptional activity, and we map the residues essential for this activity. However, since our data for some mutants showed discrepancies with previous in vitro studies, we undertook a more careful analysis of the mutant proteins using gel retardation assays and immunoblots made with nuclear extracts from transfected cells. We show that progressive C-terminal deletions drastically increase the amount of protein that accumulates in transfected cells. Immunofluorescence microscopy reveals that mutants containing between 348 and 416 residues accumulate outside the nuclear membrane, while longer mutants are nuclear like the 628 amino acid long wild type HNF1. A mutant with 289 residues is predominantly nuclear. Since the only obvious candidate for a nuclear localisation signal is located within this last mutant, we suggest that certain C-terminal deletions expose a sequence that blocks nuclear transport. PMID- 8402265 TI - [Characterization of Plasmodium falciparum antigen target of defense mechanisms developed by patients immunized against malaria]. AB - We report the identification of a 48 kDa antigen targetted by antibodies which inhibit P. falciparum in vitro growth by cooperation with blood monocytes in an ADCl assay correlated to the naturally acquired protection. This protein is located on the surface of the merozoite stage, and is detectable in all isolates tested. PMID- 8402266 TI - [Demonstration by in situ hybridization and immunohistochemistry of human insulin gene expression cells in medial habenula of transgenic mice]. AB - In transgenic mice carrying a human insulin transgene, in situ hybridization and immunohistochemistry detected a cerebral site of expression of the human insulin gene. The gene construct that was introduced into the mouse genome included 168 base pairs (bp) upstream from the transcription unit of the human insulin gene. The human gene transcript and the corresponding proteins were detected in a cell subset of the medial habenula. This was found in 20 mice from three independent transgenic lines, indicating that expression was not dependent upon the insertion site of the transgene. When the DNA fragment upstream from the transcription unit was either larger (258 bp) or smaller (58 bp), this extrapancreatic expression of the human insulin transgene was not found. This result suggests that a sequence between nucleotides -58 and -168 might be responsible for an ectopic expression of the human insulin gene, specifically in habenular cells. PMID- 8402267 TI - [High molecular weight tau proteins and acquisition of neuronal polarity in peripheral nervous system]. AB - Several variants of the microtubule-associated tau proteins, are expressed during brain development and in adulthood. These entities are required to define the polarity of the neuron and the architecture of the axon but differ in sequence and in their microtubule polymerizing activity. Here, we describe a new group of high molecular weight tau proteins that contain one or two additional exons of 711 and 198 bp in their middle region and a variable N-terminal domain. These high molecular weight tau variants are preferentially expressed in the peripheral nervous system. Immunohistochemical studies showed that they are also present in the dorsal horn of the spinal cord where they are probably transported by sensory fibers arising in the periphery. However, a minor fraction of these proteins is present in the motor neurons of the ventral horn. Similar studies were performed with the neuroblastoma N115 cell line which can be differentiated in vitro and expresses only high molecular weight tau forms. In the non differentiated cells, tau antibodies label the domain of the cell body localized around the centrosome whereas, after differentiation, the cell process facing this structure is also stained. These data suggest that axonal polarity is predetermined by the localization of tau proteins in the domain of the cell body defined by the centrosome. PMID- 8402268 TI - Neurosteroids in rat sciatic nerves and Schwann cells. AB - Pregnenolone (PREG) and corticosterone were measured in sciatic nerves and plasma of adult male rats by radioimmunoassays. Concentrations of PREG were higher in nerves than in plasma, whereas those of corticosterone were lower. In contrast to corticosterone, PREG was not reduced by castration and adrenalectomy, strongly suggesting a local synthesis of this steroid independent of glandular sources. In fact, Schwann cells in culture produced PREG from 25-hydroxy-cholesterol. They also converted PREG and dehydroepiandrosterone to the respective 7 alpha hydroxylated metabolites and to 5-androstene-3 beta,17 beta-diol. The metabolism of steroids was increased when Schwann cells were grown in the presence of insulin and forskolin. These findings may have important implications for understanding the regeneration of peripheral nerves. PMID- 8402269 TI - [Study of histidine decarboxylase activity (HDC) in murine fetal hematopoietic organs. Description of basophil-enriched splenic cell population during fetal life]. AB - The role of histamine in vitro during hematopoiesis has been described by several authors. This work was carried out to determine whether histamine could be available in hemopoietic organs by measuring the HDC activity and the histamine content of developing hemopoietic tissues from C 57 BL/6 mice during fetal life (liver from the 12th day of gestation, spleen from the 14th day, and bone-marrow from the 17th day) and postnatal life. High values were found in the liver, the bone marrow, and especially the spleen between the 17th and the 19th days of gestation. A rapid fall in these values was seen near birth. Interestingly, electron microscopy analysis of day 18 fetal spleen cells, provides evidence for a basophil-rich cell population (25%). PMID- 8402270 TI - [The presence of large acidic vesicles is correlated with in vitro invasive potential of breast cancer cells]. AB - Human breast cancer cells include large acidic vesicles (LAVs) containing endocytosed extracellular matrix (P. Montcourrier et al., Cancer Res., 50, 1990, p. 6045-6054) and mature cathepsin D, a lysosomal protease associated with metastatic risk. We show that metastatic breast cancer MDA-MB231 cells migrating in vitro through a Matrigel gel in a modified Boyden chamber were enriched in LAVs (28% compared to 6% before plating) as visualized by acridine orange fluorescence. This is the first evidence that LAVs are associated with invasiveness of cancer rather than with an apoptotic process. PMID- 8402271 TI - Modification of prostaglandin E2 and collagen synthesis in keratoconus fibroblasts, associated with an increase of interleukin 1 alpha receptor number. AB - Keratoconus, a bilateral corneal disease, is characterized by modifications in corneal shape and thinning of the stroma. It affects young people. From a biochemical point of view, a decrease in collagen content, probably due to the high collagenase activity, has been reported. In these experiments, we have studied the membrane receptors for interleukin 1 alpha, and the corresponding dissociation constant (KD). We also investigated synthesis of prostaglandin E2 (PGE2) and collagen, as well as kinetics of cyclooxygenase. Data from normal human corneas and from keratoconus were compared. Fibroblasts from keratoconus proved to bear four fold more IL1 binding sites than those from normal cornea, with similar KD. Both types of cells synthesize prostaglandin E2, even if IL1 is not added to the medium, but the keratoconus cells produce ten times more than the normal cornea cells. When the cells are stimulated with IL1, synthesis of PGE2 strongly increases and the amounts produced by keratoconus cells are always higher than those of the normal cornea cells. Kinetics of the cyclooxygenase show that Vmax. is 10 times higher in keratoconus than in normal cornea cells; Km's are the same. The amounts of collagen synthesized by keratoconus cells are slightly lower than those of normal cornea cells. Addition of IL1 to the cultures enhances synthesis of collagenase by the cells and decreases collagen found in the culture media. The drop of collagen is more important in keratoconus than in normal cornea cell cultures. PMID- 8402272 TI - [Polymerization and solubility of recombinant hemoglobins alpha 2 beta 2 6 Glu- >Ala (Hb Makassar) and alpha 2 beta 2 6 Glu-->Ala, 23 Val-->Ile]. AB - The Hb S (beta 6 Glu-->Val) fiber is formed by the packing of double strands which constitute the basic unit of the deoxyHb S polymer. The specific interaction responsible for the stabilization of the double strand involves the mutated beta 6 Val side chain (lateral contact). Recombinant Hb beta 6 Glu-->Ala and the double mutant beta 6 Glu-->Ala, 23 Val-->Ile exhibit a decreased solubility compared to Hb A. While the Hb beta 6 Ala does not polymerize, the association of the beta 23 Val-->Ile mutation at the axial contact allows the double mutant to polymerize. These results show that: (1) the hydrophobic interactions between donor and acceptor sites depend on both the hydrophobicity and the stereospecificity of the amino acid side chain at the beta 6 position; (2) increasing the hydrophobic interactions at the axial contact (connecting molecules along the same strand) result in an increased stability of the lateral contact between filaments. PMID- 8402273 TI - Acquired unbalanced hemoglobin chain synthesis during HIV infection. AB - Few abnormalities have been described up to know in the red cell lineage of HIV patients except for a slight anemia. To better interpret this, a hemoglobin study was performed in a group of 71 HIV infected patients. By contrast to patients at stage II or III, those at stage IV of the disease had moderate elevation of HbA2 level. A second group of ten patients being at stage IV of HIV infection and having moderate increase in HbA2 level were found to have beta thalassemia-like unbalanced biosynthetic globin ratio, unlike a control group with chronic inflammation condition. The observed abnormality may be a feature reflecting the peculiar disordered erythropoiesis seen in the late stage of this disease. PMID- 8402274 TI - [Ambulatory surgery: an unavoidable perspective of a partially complete concept]. AB - Whatever social security system is in place, the increasing burden of health costs on public finance and individuals own resources is difficult to bear. Available resources and equipment should be used for essential treatment and complaints about lack of resources are only justifiable when responsible use is being made of those which are currently available. Thanks to current medical techniques, new drugs and a set-up adapted to the problem, day surgery is playing a significant role in solving this problem, to the complete satisfaction of patients, surgeons and nursing teams. PMID- 8402275 TI - [A comparative study of recovery following maintenance of anesthesia with propofol or isoflurane. An attempt to synthesize current data]. AB - Quality of recovery is a means to improve anaesthetic safety during postoperative time, in post-anaesthesia recovery room and especially when the patient returns to his unity. The comparison of recovery after maintenance of anaesthesia with propofol or isoflurane shows that, for 50 to 60 min surgical procedures, results are significantly better with isoflurane. Review of literature shows that, for less than 30 min operations, propofol seems to give best recovery. For more than 30 min operations, isoflurane enables better quality recovery. PMID- 8402276 TI - [Local anesthesia and ambulatory surgery for inguinal hernia. Apropos of 737 patients]. AB - 737 cases of ambulatory herniorrhaphy performed under local anaesthesia are reported. The Shouldice technique is used. Two pharmacokinetic studies (lidocaine bupivacaine) confirm the innocuity of local anaesthesia. The authors focus their discussion on an other approach of surgery, and the medical network. PMID- 8402277 TI - [An economic analysis of ambulatory surgery of inguinal hernias]. AB - A cost-analysis study has been carried out on ambulatory surgery of inguinal hernias, in 1991, at Beziers hospital, in France. Analytical book-keeping allows to approach real costs, that is to say 3,061 FF for all medical care spending, and 2,100 FF for the hospital stay. The overall profitability of this surgery questions the urgency of its spreading within the French health care system. PMID- 8402278 TI - [Ambulatory colonoscopy under systemic general anesthesia. Our experience apropos of 500 cases]. AB - The authors report a retrospective study of 500 colonoscopies performed on outpatients under general anaesthesia. Anaesthesia protocol associated diluted propofol in continuous titrated infusion and alfentanil. The doses used were relatively low and no serious adverse effects were observed. Therefore this method appears to be appropriate for most outpatients undergoing a colonoscopy provided that usual security rules are respected. PMID- 8402279 TI - [Evaluation of ambulatory anesthesia at CHU, Brest]. AB - Outpatient anaesthesia was investigated for a two-month period by means of a questionnaire filled from the preoperative anaesthesia consultation to the surgical procedure and the discharge of the patient. 868 consultations led to schedule 260 ambulatory procedures. ENT (88 patients), paediatric surgery (73 patients) and gynaecology (63 patients) were most frequently concerned. Indications of ambulatory practice could probably be enlarged provided that recovery rooms and surgical schedules were fully adapted. PMID- 8402280 TI - [Ambulatory surgery in ophthalmology: an experience in the private sector]. AB - Outpatient surgery requires different criteria: short duration, non painful and non hemorrhagic postoperative follow up, a trained surgical and anesthesiological team. Many common ophthalmological surgical procedures meet these criteria. For a safe outpatient surgery, one must be careful for the patient selection and for the quality of the anesthesiology and the surgery. Outpatient surgery is common in other countries. In France, various centers are developing this approach. Many factors may explain why it seems easier to organize outpatient surgery in the private practice than in the public Hospital. PMID- 8402281 TI - [Locoregional anesthesia of the upper limbs. Experience of the Centre de la Main]. AB - The Hand Centre of Angers in France is entirely autonomous. 2,200 patients a year undergo hand or wrist surgery under regional anaesthesia exclusively. Authors insist on the importance of pre-selection among patients. Security rules and postoperative recommendations are mandatory. PMID- 8402282 TI - [The role of ambulatory locoregional anesthesia]. AB - The use of regional anaesthesia in day care practice is discussed. Five criteria of discharge are described: the four "A"s: awake, ambulation, alimentation, analgesia plus micturition. Complementary sedation with the regional block, if needed, should be midazolam and fentanyl. These drugs should be titrated in order to administer the efficient minimal dosage. The main techniques of regional anaesthesia are as follows: for upper limbs, intravenous regional anaesthesia, axillary and interscalenic brachial plexus blocks; for the lower limbs epidural and spinal blocks. However, urinary retention and orthostatic hypotension can occur. Furthermore the risk of headache is not a contraindication to an ambulatory practice if some guidelines are observed. In addition, penile blocks and caudal blocks are widely used in pediatrics. PMID- 8402283 TI - [Characteristic problems posed to the anesthetist by ambulatory surgery in ophthalmology]. AB - Ophthalmic surgery is one of the most valuable indications for ambulatory anaesthesia (AA). Respecting the usual recommendations for AA and the specificity of ophthalmic surgery, AA has very few problems. In USA it concerns about 90% of ophthalmic surgery. Most of the patients are very young or very old. Adults are often poly-medicamented: diabetes and arterial hypertension are the most frequent pathologies. A lot of multivisceral pathologies are responsible of ocular diseases and can complicate anaesthesia. It is necessary to diagnose them before anaesthesia. Maligna hyperthermia risk is increased during strabismus and ptosis surgery. Some ocular treatments have systemic repercussion and require to be stopped before anaesthesia. Most of ophthalmic surgery can be practiced under any types of local anaesthesia. In postoperative of strabismus and retinal detachment repair, pain, nausea, vomiting are frequently observed. Their prevention is not very well known. The atropine used for cardiac reflex treatment may be responsible of an acute urine retention or a disorientation in elderly patients and delays the home readiness. Paper and pencil tests after general anaesthesia are very difficult to do, because requesting a good vision. The postoperative complications are essentially surgical complications. PMID- 8402284 TI - [Organization pf the consultation for ambulatory anesthesia in university hospitals]. AB - The recent increase in endoscopic and radiological exploratory techniques, with the possibility of taking therapeutical action, has led to the multiplication of interventions outside the operating theatre in University Hospitals. This increase in ambulatory activity has been managed at the DAR by setting up a centralized consultation using the existing accommodation, staff and secretarial facilities. This approach has enabled us to rigourously select all patients using these facilities and to monitor their management over the last three years. The medical and paramedical staff has been rationally organized according to the posts made available. This compulsory procedure guarantees the safety of a patient who has to undergo anaesthesia. PMID- 8402285 TI - [Criteria of awakening. Street aptitude]. AB - Recovery period after anaesthesia, even if short in ambulatory surgery, must not be neglected. Most of serious accidents occur in recovery period but recovery assessments are not clearly defined. Different scores exist. Home readiness requires integrity of vital functions. Tests of upper functions also exist but are not commonly and easily used. They don't disclaim physician responsibility and increase his task appreciably. PMID- 8402286 TI - [Ambulatory anesthesia in gynecologic surgery]. AB - Surgical gynecologic procedures are among the best clinical indications of ambulatory surgery as the patients are usually young and healthy. The beneficial effects of this type of organization (economical, medical and psychological) are numerous. A rational approach of the main objectives of ambulatory surgery and anesthesia is needed to choose the more logical anesthetic agents to allow rapid patient's recovery with a minimal rate of postoperative problems. These issues are therefore briefly developed in this text. PMID- 8402287 TI - [Ambulatory celioscopy]. AB - Outpatient laparoscopic surgery implies a selection for surgical procedures in order to prevent the risk of postoperative bleeding and/or bowel injury. Anesthetists are concerned with the safety of the airway (tracheal intubation or laryngeal mask). Prevention of nausea and vomiting and analgesia are the more important points in the postoperative period. Due to the prolongation of these symptoms during 48 hours or more, the treatments must be adapted (oral route). PMID- 8402288 TI - [Prevention of nausea and vomiting in ambulatory anesthesia]. PMID- 8402289 TI - [Ambulatory anesthesia in the elderly]. AB - The aim of ambulatory anaesthesia in elderly is to reduce consequences of an interruption of their life habits. Medical and social selection criteria must be rigorous in order to obtain a great benefit comparing with short hospitalization. No previous significant study has been yet published to precise the place of this technique and its benefits for these patients. PMID- 8402290 TI - [Pediatric surgery and anesthesia in a day hospital]. AB - Day care surgery is an increasing service in our health structures. If we return to the source, we find the first important series has been published in 1906 (8,900 cases) without accident. Child is an ideal patient. So, more than 60% of paediatric surgery could benefit by ambulatory surgery. Recovery of mental abilities following general anaesthesia has not the same significance as in adult. Many studies confirm the safety of paediatric outpatient anaesthesia, but can we assert that children older than five years prefer ambulatory surgery? In the same way, are we sure that the cost cannot be cut and maybe other options used? PMID- 8402291 TI - [Professional responsibility in ambulatory techniques. The individual responsibility of the anesthetist, the surgeon and the facility director]. AB - Ambulatory practice is regulated since October 1992. These regulations do not limit directly the technical choice by surgeons and anaesthetists. However opportunity and feasibility of day care must be estimated separately by the concerned practitioners. Regulations do not affect sensibly the relation with patients though the latter are much more responsible and informed than if hospitalized. Whatever special precautionary measures are taken, aiming at a better risks prevention, they do not reduce at all the usual obligations of anaesthetists and particularly of surgeons, given the ever increasing use of recent methods such as interventional endoscopy. PMID- 8402292 TI - [Anesthesia for ambulatory surgery and anesthesia in times of crisis]. PMID- 8402293 TI - [Ambulatory surgery in otorhinolaryngology: what is possible, suitable, prohibited?]. AB - A rational examination of technical conditions in E.N.T. face and neck surgery is realized with the intention of determining the best directions of ambulatory surgery. This application is based on technical requirements due to modernization of equipment and surgical ways, specially in ear and nose surgery. In surgery of upper air and alimentary ways, patient's security is at risk. These considerations allow to discuss states in which ambulatory surgery is possible, others where it is wished and particular states where it is not recommended. PMID- 8402294 TI - [Surgery without hospitalization]. PMID- 8402295 TI - [Incidents occurring during the application of chloroform and deaths attributable to chloroform]. PMID- 8402296 TI - The future provision of orthodontic services. PMID- 8402297 TI - The use of the index of orthodontic treatment need (IOTN) as a public health tool: a pilot study. AB - Many attempts have been made in the past to develop indices of malocclusion. The index of orthodontic treatment need was developed in the United Kingdom to assess both dental health and aesthetics, primarily for clinical purposes. This study was designed to assess the use of the index of orthodontic treatment need in dental public health. Dentists in the community dental service were taught how to use the index from a hierarchical training and calibration programme. These dentists then tested the index in the field and reported back their findings. The results indicate that the index of orthodontic treatment need has potential in dental public health but some modifications are suggested. These suggestions will be incorporated into a full study alongside caries prevalence in the future. PMID- 8402298 TI - Factors affecting attitudes toward dental appearance and dental function in a Swedish population aged 45-69 years. AB - A questionnaire study was carried out on a random sample of 3,000 adults aged 45 69 years, living in Orebro county, Sweden. The response rate was 79.4 per cent. The purpose of this study was to develop measurements of attitudes toward dental function and dental appearance, and to investigate the relative importance of socio-economic factors to these attitudes. In factor analysis, two unidimensional variables explaining almost 54 per cent of the variance were developed, measuring the attitude to the importance of dental appearance and dental function. The variables were only moderately correlated. Bivariate and multivariate logistic regression models were constructed, where low education was related to high importance of dental appearance, and low socio-economic status and its interaction with education were related to high importance of dental function. Both models had low predictive values and no significant relationships were found with age, gender, marital status, place of residence, use of dental care, or to satisfaction with teeth or received care. It is concluded that it should be possible to influence attitudes to dental conditions irrespective of demographic or socio-economic factors. PMID- 8402299 TI - The use of a classification of residential neighbourhoods (ACORN) to demonstrate differences in dental health of children resident within the south Birmingham health district and of different socio-economic backgrounds. AB - This study was undertaken to evaluate variations in the dental health of children living within fluoridated South Birmingham, using a classification of residential neighbourhoods (ACORN) as a descriptor of socio-economic status, and to evaluate the change in these differences over time. Five-year-old children were examined as part of the rolling programme of epidemiological surveys co-ordinated by the British Association for the Study of Community Dentistry (BASCD) in 1987 and 1989/90. The ACORN classification of each child was determined from the postal code of the home address. ACORN groups were amalgamated into three ranked divisions. There was a variation in dental health both in 1987 and 1989/90; children from disadvantaged groups had the poorest dental health. There was more marked variation in 1987 than in 1989/90. The reduction in inequality during this period was due to a relatively greater improvement in the dental health of the children from the more socially deprived areas. PMID- 8402300 TI - Measuring social inequality in dental health services research: individual, household and area-based measures. AB - The literature on inequalities in health is extensive and provides convincing evidence that lower socio-economic groups have poorer general and dental health than higher socio-economic groups. In this literature, socio-economic status has been measured in terms of occupation, income or education. These conventional measures have a number of methodological and theoretical weaknesses which limit their ability to fully describe and explain differences in health between population sub-groups. In this paper, these measures are reviewed and alternatives, in the form of area-based indicators, are considered. The evidence suggests that measures of the socio-economic characteristics of neighbourhoods are better predictors of the health status of population subgroups than measures of the socio-economic characteristics of individuals or households. Moreover, they add additional explanatory power to models of health inequalities. Area based measures can be readily applied to survey data or information derived from health records or routinely collected sources. Since they identify where deprived groups with relatively poor health live, they may prove useful in terms of the planning and targeting of health services. These alternative indicators should be explored in the context of dentistry to determine their ability to predict differences in dental health and risk factors for dental disorders among different segments of the population. PMID- 8402301 TI - Dental caries experience in Jordanian and English schoolchildren. AB - The dental caries experience of 6- and 12-year-old children in Jordan and England was compared. The 6-year-old Jordanian children had significantly higher dmft scores than the English children, largely due to the greater numbers of decayed teeth in Jordan. Mean DMFT scores for 12-year-old children were similar in the two countries, however, the DMFT totals included more decayed and missing teeth in Jordan and more filled teeth in England. In both countries there were significant differences between the caries experience of children from different social classes. PMID- 8402302 TI - Dental caries experience and enamel opacities in children residing in urban and rural areas of Antigua with different levels of natural fluoride in drinking water. AB - In 1988/89, a national study was carried out to investigate the prevalence of caries experience among children aged 5 to 6, 12, and 15 to 19 years and also to measure the level of dental fluorosis among 12 to 14 year-old pupils attending schools in urban and rural areas in Antigua with various levels of natural fluoride in public water. In the first part of the study, all three age groups who were life-long residents of three areas with 0.1-0.2 ppmF (urban), 0.1-0.3 ppmF (rural) and 0.6-1.0 ppmF (rural) showed that the caries levels for each age group were not significantly different among the three fluoride communities or between urban and rural samples. In all three age groups, occlusal surfaces were more frequently affected by caries, and untreated dental caries was common. In part two of the survey, the Tooth Surface Index of Fluorosis (TSIF) was used to record fluorotic enamel defects among children aged 12 to 14 years who were life long residents of 0.1-0.3 ppm fluoride and 0.6-1.0 ppm fluoride areas. In the low fluoride areas, mottling was absent in 97 per cent of facial surfaces of anterior maxillary teeth. In contrast, in the 0.6-1.0 ppm fluoride area, the value was 87 per cent. In both communities mottling was limited to a whitish colour. Analysis of the highest TSIF scores revealed that statistically significant differences were apparent in children with fluorosis between two communities. PMID- 8402303 TI - Cost evaluation of two methods of post tooth extraction hemostasis in patients on anticoagulant therapy. AB - The classical management of patients on oral anticoagulant therapy included hospitalisation, cessation of the anticoagulant agent, and extraction of teeth when the prothrombine levels rise. This method was substituted in the High Risk Dental Clinic at Barzilai Medical Center in Ashkelon by use of a tissue sealant (Tisseel) which does not need hospitalisation nor cessation of the anticoagulant therapy. In comparing the last 23 sessions employing the former method to the first 23 sessions using the new method there were significant differences in the cost effectiveness for the health system, provider, insurer and patient. Despite the fact that from the health system point of view the new method is much more cost effective, there is no financial incentive for the provider (hospital) nor awareness on the part of the insurer (General Sick Fund) to embrace it and 'market' it. PMID- 8402304 TI - Assessment of enamel opacities in children in Sri Lanka and England using a photographic method. AB - Colour photographs were taken of the labial surface of both maxillary central incisor teeth of children aged 12 years, living in Sri Lanka and England. In each country, children were included who lived in communities receiving drinking water containing 0.1, 0.5 and 1.0 ppm F, and within these communities children were classed as high or low socio-economic (SE) status. The photographs were examined 'blind' by two examiners independently. These pertained to 670 children, 332 in Sri Lanka and 338 in England. The index of Developmental Defects of Enamel (DDE) was used, as modified by Clarkson and O'Mullane (1989), to measure type and extent of opacity. Intra- and inter-examiner agreement was substantial. Prevalence of opacities ranged from about a quarter of teeth in the 0.1 ppm F area in Sri Lanka to over 60 per cent of teeth in the high socio-economic group in the 1.0 ppm F area in England. Higher prevalences of opacities were recorded in: (1) the high SE group than the low SE group in the 1.0 ppm F area in England, (2) the 1.0 ppm F area than in the 0.1 ppm F area in both countries, (3) in Sri Lanka than in England in low SE groups in the 1.0 ppm F areas. The greatest differences occurred in diffuse opacities. When these data were compared with results of clinical examinations of these same tooth-surfaces by one examiner (n = 506) more teeth were graded 'normal' clinically and more teeth graded as having opacities photographically. Both demarcated and diffuse opacities were scored more frequently from photographs than clinically in both countries. PMID- 8402305 TI - Epidemiology and aetiology of oral cancer in the United Kingdom. PMID- 8402306 TI - The natural history and pathology of oral cancer and precancer. PMID- 8402307 TI - Clinical diagnostic methods for the detection of premalignant and early malignant oral lesions. PMID- 8402308 TI - Evaluation of screening for cancer. PMID- 8402309 TI - Management options in potentially malignant and malignant oral epithelial lesions. PMID- 8402310 TI - Behavioural aspects of oral cancer screening. PMID- 8402311 TI - Cost and value considerations in screening for oral cancer and precancer. PMID- 8402312 TI - Practical considerations for the establishment of an oral cancer screening programme. PMID- 8402313 TI - [Rationalization of health expenditures: the Seguin's plan in 1987]. AB - Sickness insurance is universal and compulsory in France. It is financed mainly by contributions from employers and employees. Consequently its soaring costs may sharply reduce the competitive French products on the world market. Successive French administrations have attempted to curb the growing health costs by increasing co-payment. The program launched in 1987 was innovating since it was aimed at the group of insured people who were previously not submitted to co payment (persons having presumably grave illness...). The program was a breakthrough in the French system which was built on a mixed concept of insurance and solidarity. The program was undoubtedly a great success. However, lack of an efficient information system, and reaction of the various actors (patients and doctors) who succeeded to adjust themselves to the new regulations, diminished its impact some years after it has been implemented. PMID- 8402314 TI - "Exit" and "voice" in the Italian National Health Service. PMID- 8402315 TI - [Joint support of the Association for the Right to Die with Dignity]. AB - In 1990, about 21,000 people were members of the Union. Membership is heavily dominated by women. It is noteworthy that the suicide rate is much higher among men. Moreover, young members are extremely rare. Members per 10,000 population are 3 in the age group 45-49, but the rate exceeds 19 per 10,000 among those aged 75-79. The Union was primarily established for lobbying in view of a change in the current laws. However, it seems that most of its members have in mind the quest for an answer to his/her own personal problem. Membership is more frequent in big cities than rural areas. Social control, and the activities of the Union, developed mostly in towns, are key explanations to the discrepancy. PMID- 8402316 TI - On-the-job exposure to HIV: reducing the RNs' risk (continuing education credit). PMID- 8402317 TI - Racemization of aspartic acid in the extracellular matrix proteins of primary and secondary dentin. AB - The accumulation with age of D-aspartic acid in primary and secondary human dentin was determined. In primary dentin, the plot of the D/L Asp ratio vs. age was found to fit first-order kinetics in accordance with the literature. But the secondary dentin behaved in an irregular manner and showed, in the great majority of cases, significantly increased D/L Asp ratios. Possible reasons for these findings, such as differences in protein composition and/or in the prevailing temperature, are discussed. PMID- 8402318 TI - Effects of estrogen replacement on insulin-like growth factor I concentrations in serum and bone tissue and on interleukin 1 secretion from spleen macrophages in oophorectomized rats. AB - Oophorectomy (OOX) has been known to increase bone turnover, but its precise mechanism is not fully understood. In order to further investigate the mechanism, we determined insulinlike growth factor I (IGF-I) concentrations in serum and bone tissue and interleukin 1 (IL-1) release from spleen macrophages in oophorectomized rats because it has been demonstrated that IGF-I stimulates bone formation and IL-1 stimulates bone resorption. Female 8-week-old Wistar rats were divided into four groups: (1) control, (2) OOX, (3) OOX given estradiol, and (4) control given estradiol. Ten micrograms/kg of 17 beta-estradiol was given daily by subcutaneous injection. After 5 weeks of treatment, IGF-I concentrations in the extract from right femur and in serum were determined by specific radioimmunoassay. IL-1 activity released from lipopolysaccharide (LPS)-stimulated spleen macrophages was determined by bioassay. IGF-I contents in the femur and IGF-I concentrations in serum in oophorectomized rats were significantly higher than those in control rats. Treatment by estradiol inhibited the increase in IGF I concentrations both in femur and in serum. IL-1 release from LPS-stimulated spleen macrophages in oophorectomized rats was increased, and treatment by estradiol also inhibited the stimulated IL-1 release. The ash weights and the calcium contents of left femur in oophorectomized rats were lower than those in control rats. These results suggest that both IGF-I and IL-1 may be involved in the mechanism of the regulation of bone turnover in oophorectomized rats. PMID- 8402319 TI - Effects of prednisolone and deflazacort on osteocalcin metabolism in sheep. AB - Glucocorticoids adversely affect bone and mineral metabolism through a number of mechanisms, including inhibition of bone formation. Deflazacort is a glucocorticoid which has been reported to be relatively "bone-sparing." We compared the effects in oophorectomized sheep of deflazacort and prednisolone on the metabolism of osteocalcin (OC), a marker of osteoblast function. An [125]OC infusion method was used to measure the OC plasma clearance rate (PCR) and OC plasma production rate (PPR). Six-day intravenous infusion of deflazacort and prednisolone (in the dose range 0.007-1.00 mg/hour) induced dose-dependent decreases in OC PPR which were of a similar pattern but significantly different magnitude (P < 0.02); deflazacort demonstrated a potency about 150% that of prednisolone. Both steroids decreased plasma OC levels on a dose-related basis but at the lower doses 0.05 mg/hour (P < 0.05) and 0.013 mg/hour (P < 0.0005), deflazacort caused greater decrements. OC PCR was significantly increased only by higher doses of deflazacort (1.00 mg/hour, 0.25 mg/hour; P < 0.05). Deflazacort and prednisolone increased both postabsorptive plasma glucose and plasma calcium levels, but there were no significant differences between their effects. We conclude that plasma OC levels and OC PPR in sheep were more sensitive to the effects of deflazacort than to prednisolone. At high doses, the depressive effect of deflazacort on plasma OC levels may have been due in part to an increased OC PCR which was not evident with prednisolone treatment. However, the agents appeared to have a similar dose-dependent hyperglycemic effect, and both caused a small dose-dependent increase in plasma calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402320 TI - Transforming growth factor beta 1-induced cellular heterogeneity in the periosteum of rat parietal bones. AB - We examined the osteogenesis process in transforming growth factor beta 1 (TGF beta 1)-treated neonatal and adult rats, aiming to investigate the age difference in the effect of TGF-beta 1 on mesenchymal cell differentiation. Recombinant human (rh) TGF-beta 1 (20 and 200 ng) was injected onto the outer periostea of the right side of the parietal bone of each rat once a day for 1-12 days starting at the age of either 1 day or 12 weeks. On the day after the final injection, the calvaria was excised and evaluated histologically. In the neonates, the 12-day treatment with rhTGF-beta 1 increased the number of osteoprogenitor cells, resulting in intramembranous ossification. In the adult rats, rhTGF-beta 1 induced differentiation of chondrocytes. Cartilage masses were surrounded by mesenchymal cells, which would differentiate into chondrocytes. The cartilage matrix was partially calcified, with chondrocytes buried therein. In the calcified matrix, marrow cavities containing some multinuclear osteoclasts were formed. These findings indicate that rhTGF-beta 1 stimulated the differentiation of mesenchymal cells into chondrocytes and produced the cartilaginous matrix. rhTGF-beta 1 induced intramembranous ossification of the parietal bone in neonatal rats, and it induced enchondral ossification in adults. This result suggests that the different responses of mesenchymal cells in the periosteum to rhTGF-beta 1 may depend on the age of the animals used: namely, they may reflect the respective osteogenic stages of modeling and remodeling. PMID- 8402321 TI - Electron microscopy of calcification during high-density suspension culture of chondrocytes. AB - Chondrocyte cultures grown in centrifuge tubes with intermittent centrifugation differentiate into hypertrophic chondrocytes and form calcification. We examined chondrocytes cultured in this system electron microscopically. Rat growth-plate chondrocytes were seeded in a plastic centrifuge tube and cultured in the presence of Eagle's minimum essential medium supplemented with 10% fetal bovine serum and 50 micrograms of ascorbic acid per ml. Specimens were examined by using electron microscopy and selected-area electron-diffraction techniques. In the early stage of culture, a few chondrocytes were scattered and extracellular matrices were not observed. In the middle stage of the cultures, the chondrocytes resembled proliferative cells. Matrix vesicles appeared to be budding from the cell surfaces of chondrocytes and were observed sparsely in the extracellular matrices, which were well formed around the chondrocytes. Matrix vesicles increased substantially during the following cultures. In the mature stage of the cultures, crystal formation related to matrix vesicles was observed. In the 33 day cultures, several masses of calcified matrix were formed and it was confirmed to be apatite by selected-area electron diffraction analysis. The chondrocytes appeared hypertrophic during this same stage. The 56-day culture was similar to the 33-day culture. It was concluded that this culture system provides an extracellular-matrix mineralization which is produced by chondrocytes per se. PMID- 8402322 TI - The effect of dietary xylitol on recalcifying and newly formed cortical long bone in rats. AB - Thirty-six 3-week-old male Wistar rats were labeled with a single intraperitoneal tetracycline injection. Twenty-four of them were then fed a Ca-deficient basal diet for 3 weeks, while the control group received the basal diet supplemented with CaCO3 (12 g/kg). The tetracycline labeling was then repeated and six animals in each group were decapitated. The diet of the remaining formerly Ca-deficient animals was returned to normal, and half the test rats also received xylitol supplementation (50 g/kg). After 4 weeks of rehabilitation the labeling was repeated and the animals were decapitated and their tibias were prepared. The tibias were measured in terms of weight and density and cross sections were prepared for the examination of mineral content. Bone element analysis was performed by scanning electron microscopy with electron-probe microanalysis, examining separately the bone areas formed during the various dietary periods. Areas of the former Ca-deficient and newly formed cortical bone were identified by tetracycline fluorescence under ultraviolet light, and the amount of cortical bone in each group was measured. The mineralization-promoting effect of dietary xylitol as compared with CaCO3 supplementation alone was seen more clearly in the newly formed periosteal bone than in remineralization of the formerly Ca deficient bone, the concentrations of Ca and P being significantly elevated (P < 0.05), as also was the total mineral content (P < 0.01). The cortical bone volume was similar following the CaCO3 and CaCO3 + xylitol supplementations, suggesting unaltered formation of the organic matrix. The results show that the effect of xylitol on bone during dietary Ca rehabilitation particularly concerns newly formed bone mineral.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402323 TI - The effects of liposome-encapsulated and free clodronate on the growth of macrophage-like cells in vitro: the role of calcium and iron. AB - Clodronate (dichloromethylene bisphosphonate) inhibits the activity of osteoclasts, thereby preventing bone resorption in disorders characterized by excessive bone loss. Intravenously injected clodronate encapsulated in liposomes is also known to inactivate phagocytic cells in spleen and liver in vivo. The macrophage suppressive effect of clodronate is of interest in autoimmune diseases, like rheumatoid arthritis, in which phagocytic cells are involved in inflammatory processes, but knowledge of the interaction of clodronate with phagocytic cells is scarce. We have studied the uptake of clodronate, both free and encapsulated in negatively charged liposomes, by the macrophage-like cell line RAW 264 and by other types of cell lines. The uptake was assessed by a growth inhibition assay. The liposome-encapsulated clodronate was 50 and 350 times more potent than free drug for RAW 264 and CV1-P, respectively. Cell lines with a lower endocytotic capacity were insensitive to liposome-mediated delivery of the drug. The action of free clodronate seemed to be extracellular in all cell lines studied. Calcium and/or iron have been suggested to be involved in the intracellular uptake and action of clodronate in phagocytic cells. We found that the uptake of free clodronate by RAW 264 cells was indeed mediated by calcium and iron, while the uptake of liposomal drugs was only slightly affected by calcium. The increased intracellular calcium concentration in macrophages did not significantly affect the growth-inhibitory properties of clodronate, whereas iron loading of the cells partially restored the cell growth. The data do not support the role of calcium chelation as a mechanism of action of clodronate, but suggest that intracellular iron is, at least partially involved. PMID- 8402324 TI - Health-related quality of life in osteoporosis clinical trials. The Osteoporosis Quality of Life Study Group. PMID- 8402325 TI - Beta 2-microglobulin in postmenopausal osteoporosis. AB - The so-called bone-derived growth factor, or beta 2-microglobulin, has a regulatory function in bone metabolism, stimulating osteoclastic activity. Osteoclastic activity is enhanced in postmenopausal osteoporosis, suggesting that beta 2-microglobulin concentration may also be increased in this disease. beta 2 microglobulin concentration was found to be raised (P < 0.001) in 30 women with postmenopausal osteoporosis as compared with 30 normal women of similar age; tartrate-resistant acid phosphatase concentration also was raised (P < 0.001), and total body bone mineral content was decreased (P < 0.001). Linear regression analysis revealed a highly negative correlation result between total body bone mineral content and beta 2-microglobulin (r = 0.577, P < 0.001), and a positive correlation result between beta 2-microglobulin and tartrate-resistant acid phosphatase concentration (r2 = 0.806, P < 0.001). These findings, and the stimulatory effect of beta 2-microglobulin on osteoclastic and osteoblastic activity, suggest that beta 2-microglobulin may play an important role as a local regulatory factor in the pathogenesis of postmenopausal osteoporosis. PMID- 8402326 TI - Vitamin K-induced changes in markers for osteoblast activity and urinary calcium loss. AB - The objective of this study was to identify subjects in whom vitamin K has an effect on markers for calcium and bone metabolism and to detect hitherto unnoticed correlations between vitamin K-induced changes in these markers. Participants in our studies were apparently healthy women, in whom we measured serum-immunoreactive osteocalcin (irOC) before and after adsorption to hydroxylapatite; total serum alkaline phosphatase (T-AP) and bone-specific alkaline phosphatase (B-AP); and fasting urinary calcium and creatinine. We describe a trial among 145 women who were treated with vitamin K (1 mg/day) for 2 weeks, and a prospective placebo-controlled trial among two groups each of 70 postmenopausal women with a treatment period of 3 months. It turned out that in elderly women vitamin K induced increased levels of serum irOC with a high affinity for hydroxylapatite (irOCbound), whereas that with low affinity (irOCfree) remained unaffected. In placebo-treated women the ratio irOCfree/irOCbound shifted from 0.38 to 0.65 around the 50th year of age. This shift was not found in vitamin K-treated women. After 3 months of treatment the vitamin K-induced changes in irOCbound were correlated with changes in B-AP, whereas irOCfree was correlated to urinary calcium excretion. In fast losers of urinary calcium vitamin K induced a 30% decrease of calcium excretion. The hypothesis is put forward that irOCbound may be a marker for bone formation, that serum irOCfree may be a marker for bone resorption, and that the serum irOCfree/irOCbound ratio may become a marker for skeletal remodeling.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402327 TI - Serum osteocalcin and fracture susceptibility in elderly women. AB - The purpose of this investigation was to evaluate the bone turnover by using bone metabolic markers in relation to previous fracture history and independent of bone mass. Patients and controls were recruited from a population-based study of 193 women, all living in the same city and aged 60, 70, and 80 years. The bone mineral content (BMC) was measured bilaterally in the distal forearm by single photon absorptiometry (SPA). At the same time, serum samples were obtained for biochemical analysis. Of the 193 women, we identified 26 with at least one major fracture during the past few years. Each of these 26 women with a certified recent previous fracture was individually matched with a woman from the same study group of equal BMC and age but without a fracture history. In the two groups, the serum samples were analyzed for osteocalcin, C-terminal procollagen peptide (P1CP), alkaline phosphatase, bone-specific alkaline phosphatase, calcium, phosphate, and albumin. The serum concentration of osteocalcin was 20% lower in the women with a previous fracture than in the controls (P = 0.03). The other markers of bone formation gave similar values in the two groups. There was a significant correlation between the osteocalcin and P1CP concentrations (P = 0.001). Our findings indicate that the susceptibility to fractures independent of factors such as age and BMC may be related to a decreased bone turnover. PMID- 8402328 TI - Localization of estrogen receptors in long bones and vertebrae of human fetuses. AB - In order to investigate the possible role of estrogen in the development of cartilage and bone we studied by immunofluorescence immunohistochemistry and autoradiography 26 human embryos and fetuses 7-22 weeks in gestational age associated with pregnancy interrupted for non-medical reasons. In order to demonstrate the presence of estrogen receptors (ERs) in human fetal cartilage, cryostat sections of long bones and lumbar and thoracic vertebrae were prepared for (1) fluorescent immunocytochemistry using an antiidiotypic monoclonal antibody to anti-estradiol receptor monoclonal Ab labeled with fluorescein isothiocyanate (FITC), (2) immunohistochemistry using monoclonal antihuman estradiol receptor antibody, labeled with strept. A-B immunoperoxidase, and (3) autoradiographic localization of estradiol using labeled (3H) 17 beta estradiol. In fetuses aged 10 weeks or older, intranuclear and perinuclear localization of ER was demonstrated by all methods, mainly amongst chondrocytes of the proliferating and higher hypertrophic zones of the epiphyses and in the cartilage of vertebral bodies. These data suggest that estrogen acts directly on chondrocytes of human fetuses through an ER-mediated mechanism. PMID- 8402329 TI - Insulinlike growth factor gene expression in human fracture callus. AB - The effects of insulinlike growth factors on bone and cartilage-derived cells in culture have been extensively investigated, but there is little information on their role in vivo in bone, especially in fracture healing. This study investigated insulinlike growth factor (IGF) I and II mRNA expression in normally healing human fractures by in situ hybridization. Endothelial and mesenchymal cells at the granulation tissue stage expressed IGF-II mRNA. At the stage of bone and cartilage formation, osteoblasts and non-hypertrophic chondrocytes expressed mRNA for both IGF-I and II. Some osteoclasts were positive for IGF-II mRNA at the stage of bone remodeling. The greater time span of IGF-II expression relative to IGF-I reflects the predominance of IGF-II in human bone matrix. Taken together with the known effects of IGFs on bone and cartilage cells in vitro, these findings support a role for IGFs in local cellular regulation in human fracture healing. PMID- 8402330 TI - Socialized dentistry. PMID- 8402331 TI - U.S. board exams. PMID- 8402332 TI - HIV/AIDS, quirks and quarks. PMID- 8402333 TI - Management of a dental practice. PMID- 8402334 TI - Dr. C. Edmund Kells, Jr.--dental genius. PMID- 8402335 TI - Refusal to replace amalgams does not constitute malpractice. PMID- 8402336 TI - Choosing insurance in step with your needs: the advantages of step-rate over level premiums. PMID- 8402337 TI - Determining your practice's business value. PMID- 8402338 TI - Meet the Wenns from P.E.I.--a true dental family!. Interview by P.R. Crawford. PMID- 8402339 TI - Fractures of the zygomatic complex: a case report and review. AB - Traumatic injuries to the midface are not nearly as common in Canada as they are in the United States, but practitioners must still be prepared for the evaluation and treatment of patients presenting with midface fractures. This case report demonstrates the experience one individual who presented with the clinical signs and symptoms of midfacial trauma. Cases of this nature can be confidently managed by an oral and maxillofacial surgeon. A review provides knowledge regarding anatomic features, classification schemes and the diagnostic and therapeutic decisions encountered with the treatment of zygomatic complex fractures. The forward projection of the zygoma causes it to be frequently injured secondarily to blunt trauma of the midface, at the expense of protecting the orbit. PMID- 8402340 TI - Pediatric mandibular fractures treated by rigid internal fixation. AB - Mandibular fractures in the pediatric patient population are relatively uncommon. These patients present with their own unique treatment requirements. Most fractures have been treated conservatively by dental splints. Closed reduction techniques with maxillomandibular fixation (MMF) in very young children can pose several concerns, including cooperation, compliance and adequate nutritional intake. Rigid internal fixation of unstable mandibular fractures using miniplates and screws circumvents the need for MMF and allows immediate jaw mobilization. At major pediatric trauma institutions, there has been an increasing trend toward the use of this treatment when open reduction is necessary. This article presents a report of a five-year-old child who presented with bilateral mandibular fractures and was treated by rigid internal fixation and immediate mandibular mobilization. PMID- 8402341 TI - Clinical observations on a newly designed electronic apex locator. AB - The introduction of alternative electronic principles and circuitry has produced the next generation of electronic apex locators, which have fewer clinical limitations and greater ease of use, on a routine basis, than their predecessors. The history of how the electronic apex locator was developed over the past 30 years and the limitations of the machines designed on earlier principles are reviewed. Clinical examples of root canal systems measured using a newly designed, frequency-dependent apex locator are presented in order to illustrate the spectrum of cases that can be read. All measurements were made on canals irrigated previously with electro-conducting solutions, which would have made readings from earlier machines problematic. Single-rooted and molar teeth as well as teeth with vital tissue in the root canal system or with necrotic pulp tissue and periapical lesions have been treated with equal ease and results. The electronic apex locator is being advocated to supplement instrumentation technique and to improve the identification of the apical terminus of the root canal system. The routine use of these devices is advocated to ensure an accurate placement of root canal fillings rather than to eliminate the need for radiographs. PMID- 8402342 TI - [Comparative study of the point diameter of dental explorers and the effect of wear on it]. AB - The diameter of dental explorers from three companies have been measured at 10, 20, 40 and 50 microns from the end of the tip, first on new explorers and then again after submitting these explorers to a stimulated wear test of 25 and 150 times. The study has shown that on new explorers Hu-Friedy had the most uniform tip diameter. The tip diameter of explorers from Healthco and Hu-Friedy were finer then Premier's, and Premier explorers seemed to show the least wear. PMID- 8402343 TI - Staff/student relations: building the trust. PMID- 8402344 TI - [The grafting of mixed pulp of autoepidermis and autodermis in full-thickness skin burns]. AB - This article described the procedures of grafting of mixed pulp of autoepidermis and autodermis in 22 cases of extensive third degree burns. It solved the problem of the shortage of autoskin. The proportion of donor site to wound surface reached 1:24. As the mixed skin pulp contained a large quantity of autodermis, the wound surface healed well without rejection. PMID- 8402345 TI - [Clinical study on main visceral damage and multiple organ failure (MOF) following severe burns]. AB - A prospective study was carried out on 57 patients with total burned surface area (TBSA) over 30%. It was found that myocardial damage occurred early postburn, which was one of the major causes of cardiac dysfunction and failure. The postburn respiratory failure (RF) might be classified into three patterns. The etiology of each pattern varied. The imbalance between thromboxane and prostacyclin in plasma and visceral tissues played important roles in the genesis and development of postburn MOF as well as the causes of pathophysiological alterations in the main factors (including inhalation injury, severe shock and systemic infection) which contributed to occurrence of visceral damage and MOF. PMID- 8402346 TI - [Relation between energy intake and expenditure in Chinese burn adults]. AB - In order to understand the relation of energy intake and expenditure after burn, calorie intake and resting energy expenditure of 75 burn adults with 5-98% TBSA (Total Body Surface Area) were investigated from PBD1 (Postburn Day) to PBD28. Results were as follows, 1. Burn adults with < 30% TBSA could acquire enough calorie by oral intake only, it was not necessary for them to receive special nutritional treatment; 2. Patients with 30-50% TBSA should be given extra calorie supplement during PBD1-14, because they could not get sufficient energy by oral intake only during this period; 3. It was very necessary to give patients with 50% TBSA, especially > 70% TBSA active calorie supplement, because the calorie taken by oral route during PBD1-PBD28 was not enough to satisfy the need of burn metabolism. PMID- 8402347 TI - [Detection and analysis of the red cell immune function, fibronectin, complement 3 and complement 4 in patients with burn injury]. AB - The immune function and many other physiological functions are severely interfered with in patients following burn injury. This paper reports the results of determinations of the red cell immune function, the serum concentration of fibronectin (Fn), C3 and C4 in patients with burn injury, whose third-degree burns covered 11.48 +/- 5.80%. The rosette rate of red cell C3b receptors in the patients and normal individuals were 14.28 +/- 4.58% and 17.33 +/- 4.07% (P < 0.001), and that of red cell immune complex (IC) 4.68 +/- 1.77% and 4.94 +/- 1.26 (P > 0.05), the serum Fn level 234.28 +/- 98.05 mg/L and 312.50 +/- 70.99 mg/L (P < 0.01), C3 2014.24 +/- 438.40 mg/L and 1960.00 +/- 613.30 mg/L (P > 0.05) degrees C4 601.11 +/- 84.71 mg/L and 598.24 +/- 111.09 mg/L (P > 0.05), respectively. In the patients, the rosette rate of red cell IC and the serum concentration of C3 and C4 did not change remarkably, but the rosette rate of red cell C3b receptors and the serum fibronectin level were reduced markedly in comparison with normal controls. The decrease of the last two was related to the failure of anti-infectious capacity of patients with burn injury. PMID- 8402348 TI - [Biochemical analysis of postburn scar]. PMID- 8402349 TI - [Experiences in the diagnosis and treatment of systemic fungal infection in patients with extensive burns]. PMID- 8402350 TI - [Comparison of therapeutic value of various topical agents in Pseudomonas aeruginosa infection. An experimental study]. AB - 40 Wistar rats were scalded by boiling water for 12 seconds resulting to 10% TBSA full-thickness burn. Pseudomonas aeruginosa (10(9)/ml) was smeared on the burn wounds 10 min after injury. The animals were divided into 4 groups. Sulfadiazine silver and sulfadiazine zinc (with 5% azone) cream was applied to the wounds of animals in the group 1. The contaminated wounds were irradiated with negative ion current in the group 2. The "moisture ointment" was applied to the wounds in the group 3. Group 4 consisted of the controls without any treatment. All the topical agents were applied to the wounds 4 hr after the injury. The number of bacteria in the subeschar tissue was determined every day for 4 days. The tissues of lung, kidney, liver and heart were examined pathologically on the 4th day. The results showed that the quantity of bacteria in the subeschar tissue was significantly less, and pathological changes in organs were significantly milder in the 1st and 2nd groups than other two groups. The "moisture ointment" group yielded the poorest results in respect to subeschar bacterial counts and visceral pathology. PMID- 8402351 TI - [Bacteriostatic effects of silver norfloxacin, silver sulfadiazine, and "moisture burn ointment"]. PMID- 8402352 TI - [Patho-morphological changes in the brain after severe burns]. AB - This paper reports the dynamic patho-morphological changes of brain in rats within one month after severe extensive burn (30% TBSA, II degrees-III degrees) by light microscope, electron microscope and histochemistry. The brain tissues exhibited conspicuous and consistent morphological changes. They were characterized by: (1) swelling of endothelial cells associated with increase in number of pinocytotic vesicles, tubular structure, formation of endothelial gap; (2) degeneration, necrosis, and swelling, decreased or disappearance of organelle in neuron and neuroglia cells; (3) perivascular neuroglia cuffs; (4) neuronophagiod phenomenon. The lesions appeared at 6 hr postburn, and it became more marked with elapse of time and became prominent at 3d postburn. Most of the lesions were mild and disappeared at 30d postburn. The significance and pathogenesis of the main lesions were discussed. PMID- 8402353 TI - [Changes in arachidonic acid metabolic state after burns and their significance]. PMID- 8402354 TI - [Reconstruction of unilateral maxillary defect with composite osteomuscular flap consisting of mandibular ramus]. AB - Temporo-mandibular ramus osteomuscular flap (TMROMF) was used for the reconstruction of maxillary defect after unilateral maxillectomy in 12 patients, with success in 11. Thus, zygomaticofacial deformity, naso-oral fistula, speaking dysfunction and difficulty of denture reestablishment were prevented. 10 patients are able to resume their previous work and proper social activity after the operation. TMROMF is easier to perform with few serious complications. PMID- 8402355 TI - [Surgical correction in one stage of deformities of both mandible and maxilla]. PMID- 8402356 TI - [Free transplantation of composite lip tissues. A report of 13 cases]. PMID- 8402357 TI - [Full-thickness advancement flap of cheek with muscular pedicle in repair of defect of lips]. AB - This paper reports a new technique to repair defects of the lip. It has been done in nine patients (2 upper lips and 7 lower lips), and the resultant oral configuration and function are satisfactory. The design and procedure are described in detail. The advantage of the method, the point for attention in performing the operation, and some problems in the repair of vermilion are discussed. PMID- 8402358 TI - [The third dorsal metacarpal artery neuromuscular island flap]. AB - This paper describes the usefulness of the third dorsal metacarpal artery neurovascular island flap in the repair of soft tissue defect of the hand. The flap is based on the third dorsal metacarpal artery as its vascular pedicle, and it has been successfully used in 12 cases since 1990. We found the third dorsal metacarpal artery was larger and more constant than the first, and so the blood supply was highly guaranteed. There were no failures or donor site complications. However, the flap has its limitation. It is only suitable for covering relatively small defects. The anatomy and clinical dissection of the flap are described. PMID- 8402359 TI - [Applied anatomy and clinical application of the reverse dorsal metacarpal flap]. AB - The dorsal metacarpal vessels contribute to the fascial plexus which supplies the skin of the dorsum of the hand. We have successfully used 11 reverse dorsal metacarpal flap based on the dorsal metacarpal arteries. The design and use of the reverse dorsal metacarpal flap are described. PMID- 8402360 TI - [Applied anatomy of the second dorsal metacarpal artery island flap]. AB - 50 specimens of the adult hand were dissected to study the origin, course, anastomosis and supplying area of the second dorsal metacarpal artery. The island fasciocutaneous flap of dorsum of the hand, based on second dorsal metacarpal artery, has been designed for the repair of soft tissue defects in the hand. We have successfully used four neurovascular flaps. The anatomy and clinical use of the axial pattern flap based on the second dorsal metacarpal artery are described. PMID- 8402361 TI - [Transsexualism]. PMID- 8402362 TI - Epstein-Barr virus encephalitis: clinical observations in nine Chinese children. AB - Nine Chinese children (four males, five females) in whom serology showed Epstein Barr virus infection were studied retrospectively. They were from 1.5 to 14 years of age. Various symptoms and signs, including disturbance of consciousness, visual hallucination, cranial neuropathies and sphincter dysfunction, led to initial clinical impressions of encephalitis, encephalomyelitis and neuro degenerative disease. Electroencephalography showed focal spikes over the frontal, occipital, temporal and parietal areas. The patients underwent brain computed tomography and/or magnetic resonance imaging, and three of them underwent HMPAO SPECT. Eight patients appeared normal neurologically during follow-up, and the single patient who had spastic quadriparesis improved gradually. Though visual hallucination is not a specific presentation, it is a peculiar picture of Epstein-Barr virus encephalitis. Epstein-Barr virus encephalitis seemed to be self-limiting without specific treatment in this group. PMID- 8402363 TI - [Pulmonary cryptococcosis: analyses of 17 cases in VGH-Taipei]. AB - Seventeen patients with pulmonary cryptococcosis collected at VGH-Taipei from 1967 to 1991 were analysed. Their ages ranged from 16 to 70 years with a mean of 45.2. Twelve were men and five were female. The chief characteristic symptoms and signs were cough (58.8%), fever (29.4%) and dyspnea (23.5%). Nearly one third of cases were asymptomatic and presented with abnormal chest roentgenogram. The chest roentgenogram included single nodule or mass (41.2%), multiple nodules (35.3%), pneumonic patch or consolidation (23.5%). In addition, 5 cases (29.4%) and 4 cases (23.5%) were combined with cavitary formation and pleural effusion respectively. Only two cases had associated immunologic deficiency. No patient was found to have history of breeding pigeon. Five cases didn't receive any treatment but were followed from 6 months to 5 years. The remaining 12 cases underwent medical treatment, surgical resection or a combination of both. Prognosis was good in all except one patient who died of multiple myeloma. PMID- 8402364 TI - [The effect of intracranial pressure caused by airway suction in hypertensive intracerebral hemorrhagic comatose patients]. AB - Ten hypertensive intracerebral hemorrhagic comatose patients in the Neurosurgical Intensive Care Unit were involved in this study. The ten were within the range of 5 to 8 by Glasgow Coma Scale, and all had undergone tracheostomy. During suction of the airway for 20 seconds in 4 areas (trachea, bronchus, nasal and oral cavities), the data of intracranial pressure (ICP), arterial blood pressure, pulse rate (PR) and respiration rate (RR) were recorded. Results were analyzed for ICP, mean arterial blood pressure (MABP), PR, RR, and cerebral perfusion pressure (CPP) at the 5th, 10th, 15th and 20th seconds in the above-mentioned areas. The ICP was found to be significantly increased, and above the 20 mmHg mark, during suction of the bronchus or nasal cavity for 5 seconds. The MABP also showed significant increase, to above the 110 mmHg mark during 5-second suction of the trachea or nasal cavity or during suction of the bronchus or oral cavity for 10 seconds. However, during the suction of the four airway areas, PR and RR were less influenced and the CPP was not decreased or significantly changed. The conclusion of this study was that suction in the areas of trachea, bronchus and nasal cavity, should not exceed 5 to 10 seconds; in the oral cavity, it may be done within 10 to 15 seconds and at each procedure of airway suction. If it is not possible to clean the sputum or secretion in one procedure, the patient should be allowed to rest for about one minute or become stabilized as determined by watching the monitors of ICP, BP, PR, and RR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402365 TI - Abnormal mitochondria in Rett syndrome: one case report. AB - A 6-year-9-month-old girl with the characteristic features of Rett syndrome is reported. Clinically, she had microcephaly, psychomotor arrest, deterioration of communication, autistic behaviour, loss of language development, gait apraxia and stereotyped hand washing movement. Amino acid and organic acid analysis were normal. An abnormal rise in serum lactate was noted 120 minutes after oral glucose loading. Muscle biopsy was performed and there was no specific finding noted under light microscope. Electron microscopic evaluation revealed mild accumulation of mitochondria at subsarcolemmal area with abnormal tubular cristae. The cause of Rett syndrome remains obscure. Several articles concerning abnormal mitochondrial morphology or respiratory enzymes have been reported. The exact pathogenesis requires further investigation. PMID- 8402366 TI - Adult respiratory distress syndrome caused by pulmonary cryptococcosis in an immunocompetent host: a case report. AB - The development of adult respiratory distress syndrome (ARDS) can be associated with a variety of clinical disorders. Pulmonary cryptococcosis occurring in immunocompromised patients has been reported with increasing frequency because of the rapidly rising number of immunocompromised hosts and the improvement in diagnostic techniques. But pulmonary cryptococcosis causing ARDS in immunocompetent patients has not, to present knowledge, been described. Here a rare case of pulmonary cryptococcosis is reported in an immunocompetent host who developed adult respiratory distress syndrome. The clinical course, radiologic patterns, methods of diagnosis and treatment are reviewed. PMID- 8402367 TI - Early extramedullary relapse of acute myelogenous leukemia in bilateral breasts: a case report. AB - We report a case of a 19-year-old female with acute myelogenous leukemia successfully treated with daunorubicin and Ara-c. Eight months after complete remission, palpable breast masses developed bilaterally. Biopsy and fine needle aspiration of the right mass showed cluster of leukemic cells indicative of extramedullary leukemic relapse. One month after biopsy, the patient's bone marrow showed presence of 90% blast cells. Palpable breast mass may be an early sign of relapse of acute myelogenous leukemia. PMID- 8402368 TI - Recurrent pneumonia resulting from retained esophagus following esophageal replacement for corrosive stricture: a case report. AB - Benign corrosive stricture of the esophagus rarely requires esophageal replacement due to failed dilatation. A patient is presented with severe esophageal stricture from corrosive injury; the native esophagus was eventually replaced with an ileocolon interposition graft. He suffered from recurrent pneumonia one year after operation. Mucocele formation from the retained esophagus with compression of the tracheobronchial tree, was diagnosed by computerized tomographic (CT) scan, and was resected. The clinical status improved dramatically after the procedure. Tracheobronchial compression by mucocele from the retained esophagus should be considered in the differential diagnosis of recurrent pneumonia after esophageal replacement. PMID- 8402369 TI - [Chloroform-induced hepatic injury: a case report]. AB - Chemical hepatic injury is not rare in Taiwan. We here report such an injury induced by chloroform. A previously healthy 26-year-old married woman was admitted under the impression of acute chemical hepatitis in December, 1991. Two weeks before, she had used chloroform as glue to unite two pieces of plastic pencil sharpener at home. Nasty odor was sensed while working. After series of hepatitis workup, liver echo, site visit, survey of coworkers and gas chromatography for the "glue", the diagnosis of chloroform-induced hepatic injury was confirmed. This case found that neither employer nor employee was aware of the toxicity of chloroform. No ventilation system or personal respiratory protection equipment was provided. Preemployment training and Material Safety Data Sheet were obviously insufficient, also. The diagnostic criteria of chemical hepatic injury were therefore proposed to alert medical professionals, industrial hygienist, safety personnel and factory inspectors, not to prompt an early diagnosis but also hopefully to prevent the occurrence of occupational liver diseases. PMID- 8402370 TI - Five-year experience of human immunodeficiency virus type 1 national screening program implemented at Veterans General Hospital-Taipei. AB - From July 1986 through June 1990, 33,199 sera from various risk groups were collected in Veterans General Hospital-Taipei for detection of antibody against human immunodeficiency virus, type 1 (HIV-1). Sixty-five samples were proved positive by Western blot analysis. Among individual high risk groups, hemophiliacs had the highest positive rate of 20/60 (29.41%), followed by homosexual/bisexual males (41/1,264, 3.24%). The overall positive rate was 65/33,199 (0.19%). Ten cases were recognized as acquired immunodeficiency syndrome (AIDS), 1 case had AIDS-related complex (ARC) and 4 case had other apparently symptomatic infections. Among these 15 cases, 7 expired, 1 lost of follow-up and 7 surviving cases are being treated with zidovudine (AZT). Most of symptomatic HIV-1 antibody positive cases had abnormal T4/T8 ratio of 0.39 +/- 0.54 as compared with the asymptomatic HIV-1 carriers at a ratio of 0.81 +/- 0.69. The opportunistic infections included Pneumocystis carinii pneumonia (PCP) in 6 case, disseminated cytomegalovirus infection in 6 cases, herpes zoster virus infection in 3 case, candidiasis in 4 cases, syphilis in 3 cases, pulmonary tuberculosis in 2 cases, and others with cryptococcosis, salmonellosis, Mycobacterium avium-intracellulare infection, gonorrhea, Staphylococcus aureus endocarditis and bacterial sepsis, etc. The natural history of HIV-1 infection to AIDS involved acute and persistent multiple infections. Although prevalence of HIV-1 infection was low in Taiwan, nationwide surveillance of HIV-1 infection in various risk groups is still needed. PMID- 8402371 TI - Endoscopic examination in patients with acquired immunodeficiency syndrome: Taiwan experience. AB - Endoscopic experience in patients with acquired immunodeficiency syndrome (AIDS) has rarely been reported in Taiwan. We present our experience in 9 AIDS patients (8 male and 1 female, age from 26 to 63 years) with 12 examinations. The risk factor of these patients were bisexual in 3, homosexual in 2, hemophilia in 1, drug abuse in 1, and paid-sex in 2. Odynophagia or dysphagia was the major complaints. Oral ulcers or/and thrush were noted in 8 patients. Endoscopic findings included negative (6/12), candidiasis (3/12), erosions (1/12), ulcers (1/12) and ulcer scar (1/12) in esophagus; negative (8/12), gastritis (1/12), erosions (1/12), ulcers (1/12) and Kaposi's sarcoma (1/12) in stomach; and negative (11/12) and duodenitis (1/12) in duodenum. Patients with esophageal candidiasis always had oral thrush. Dysphagia was highly correlated with positive endoscopic findings in esophagus. It is important for an endoscopist to identify clinical symptoms and to examine patient's oral cavity before an endoscopic examination. The endoscopist must keep himself from being infected by exposure to contaminated blood and secretion and avoid dissemination of this horrible disease by undisinfected instruments. PMID- 8402372 TI - Prenatal cytogenetic diagnosis by amniocentesis before 15 weeks' gestation. AB - Three hundred and ten early amniocenteses before 15 weeks' gestation, including one set of triplets and four twins, were performed for prenatal cytogenetic diagnosis. There was a 100% success rate in obtaining amniotic fluid and 99.4% success rate in achieving amniotic cell cultures. The mean time from amniocentesis to cell harvest by the in situ culture method was 11.3 days. A total 13 abnormal fetal karyotypes were diagnosed. The total fetal loss rate (spontaneous abortion, elective termination and late loss) was 4.5%, while miscarriage within two weeks after amniocentesis occurred in three cases (1%). Early amniocentesis combined with in situ cell culture is a safe, rapid and effective technique for early prenatal cytogenetic diagnosis. PMID- 8402373 TI - Continuous intravenous infusion of leucovorin and 5-fluorouracil for the treatment of advanced colorectal and breast cancers. AB - The efficacy of the protracted infusion of 5-fluorouracil (FU) 200 mg/m2/day admixed with leucovorin (LV) 20 mg/m2/day was assessed for the treatment of advanced colorectal and breast cancer patients. Of 19 patients who underwent heavy treatment, 4 patients demonstrated partial response, 13 become stable and 2 failed to respond to therapy. The overall duration of response ranged from 1.5 to 15.8 months (median 4.5 months) and the survival ranged from 1.7 to over 21.7 months (median 8.2 months). The toxic effects, specifically mucositis, oral ulcer and acral hyperpigmentation, were mild to moderate in grade. Myelosuppression with grade I leucopenia was observed in all the patients. Catheter-related complications such as thrombosis or infection occurred in 4 patients. No death relating to therapy was reported in this study. Continuous intravenous infusion chemotherapy with an admixture of LV and FU is not associated with severe toxic effects when treatment is closely monitored and appropriately administered. This treatment regimen may have some significant benefit in advanced colorectal and breast cancer patients. PMID- 8402374 TI - Pigmented villonodular synovitis. AB - Seventy-three patients, 28 males and 45 females, diagnosed with pigmented villonodular synovitis (PVNS) between 1982 and 1990 were followed for an average of 42.2 months (range, 21 to 108 months). Fifty-five cases occurred in the tendon sheath and eighteen in synovial joint. Ages ranged from 5 to 77 years, with an average of 38.9 years. The clinical presentations included chronic tumescence, serosanguinous arthrocentesis and painful limitation of range of motion. Plain roentgenography usually demonstrated soft tissue swelling with varying density, bony erosion or joint space narrowing. Arthrography and magnetic resonance imaging both gave specific pictures, with good diagnostic rates. The therapeutic regimen consisted of marginal excision for tendon sheath lesions and total synovectomy for intraarticular lesions. Extensive synovectomy with prosthesis replacement yielded good functional results in elderly patients with PVNS and cartilage destruction. Recurrence rate averaged 26% and was higher among knee joint lesions (33%), possibly because of inaccessibility of the popliteal fossa and inadequate excision of the lesion. Differences in the mitotic index between recurrence and non-recurrence groups had no statistical significance. This phenomenon may well serve against "tumor origin" as the etiology of this disease. PMID- 8402375 TI - Driving down the electronic highway. AB - Life as a computer lab coordinator is never dull nor routine. No sooner do you become comfortable with computer-aided instruction (CAI), computerized testing, interactive video systems, and local area networks, than do you face a new challenge. As faculty and students become more sophisticated in learning how to use computers, interest in accessing large, international computer networks increases. This article describes the available resources, and both the rewards and problems associated with preparing yourself to teach nursing students about the Internet, a large, global association of government, academic, and research computer networks. PMID- 8402376 TI - Imaging: an innovative technology. AB - Imaging systems combine several technologies to bring the visual dimension to the information age. Computer literature identifies imaging technology as one of the top five fastest growing technologies for the 1990s, with a projected growth rate of 54% and revenues of $6.8 billion by the end of 1993 (Runyan, 1989). Acceptance and use of digital imaging systems have grown because of the maturation of some key technologies which underlie image processing and the increased demand for image and graphic applications in the commercial and scientific sectors. This article presents the historical development of imaging technology, along with a brief description of how the technology works. Current limitations and future possibilities of the digital imaging process are presented. Applications and implications of this technology also are discussed. PMID- 8402377 TI - On-line access to nursing literature. AB - Providing access to the multiple databases essential to nursing practice and education is a challenge for universities and health care providers alike. Collaboration between Wayne State University and other institutions in the Detroit metropolitan area provides a unique system for user access to nursing information. Building on the Detroit Area Library Network (DALNET) structure, additional computer databases have been added to the Library User Information System (LUIS) to allow faculty and student access from remote locations. With university identification, faculty and students access up-to-date literature searching tools and check the availability of desired journal articles at multiple locations throughout the large tri-county area. PMID- 8402378 TI - A comparative assessment of interactive videodisc instruction. AB - A counter-balanced design was used to compare student knowledge and self-efficacy following interactive videodisc (IVD) and traditional instruction on intravenous therapy procedures. Students were assigned randomly to treatment groups receiving either IVD instruction followed by traditional lecture or lecture followed by IVD. Subjects were pretested before any instruction and post-tested after each type of instruction to provide three repeated measures for both knowledge and self-efficacy. While both types of instruction produced significant gains over baseline observation, neither method, when used by itself, was clearly superior. Combining both methods, regardless of sequence, was associated with maintained or enhanced knowledge and self-efficacy scores. Traditional-lecture presentation first, followed by IVD instruction, produced the largest gains in self-efficacy for applying intravenous procedures in nursing practice. PMID- 8402379 TI - Mail by modem: the BITNET connection. AB - Nursing faculty in university settings are now able to communicate with colleagues around the country or in other parts of the world through electronic networks such as BITNET. This article describes how members of the Southern Nursing Research Society's (SNRS) Governing Board used the BITNET system for accomplishing the society's work. Essential hardware and software requirements for using the network are described. Guidelines for logging on, sending messages, and e-mail courtesy are explained. Examples are included for using the network for 1) research and manuscript collaboration; 2) organizational communication and planning; and 3) teaching activities. PMID- 8402380 TI - Bedside/point-of-care technology: the 'window' into the integrated clinical database, Part 5. PMID- 8402381 TI - Effect of erythromycin on D-galactose absorption and sucrase activity in rabbit jejunum. AB - Erythromycin, an antibiotic used in the treatment of infectious diseases, produces gastrointestinal side effects such as diarrhea. The mechanisms by which erythromycin produces these effects are not known. However, erythromycin has been shown to increase gastrointestinal motor activity and to inhibit intestinal neutral amino acid absorption. Both effects could contribute to the gastrointestinal side effects observed. Because the intestinal systems of amino acid and sugar transport present similar characteristics, the aim of the present work was to determine whether erythromycin also alters D-galactose absorption and sucrase activity in rabbit jejunum. The results show that erythromycin diminishes intestinal D-galactose absorption. This effect seems to be due to an action mainly located on the Na(+)-dependent sugar transport of the mucosal border of the intestinal epithelium. Erythromycin also inhibits the Na(+)-K+ ATPase activity of the enterocyte, which might explain the inhibition of the D-galactose Na(+)-dependent transport. However, a direct action of the erythromycin molecule on the Na(+)-dependent carrier cannot be excluded. Erythromycin did not alter sucrase activity. PMID- 8402382 TI - Influence of atrial natriuretic factor on uptake, intracellular distribution, and release of norepinephrine in rat adrenal medulla. AB - Several studies have demonstrated that atrial natriuretic factor can bind to adrenal medulla cells. Furthermore, atrial natriuretic factor immunoreactivity has been identified in chromaffin cells. The aim of the present work was to investigate atrial natriuretic factor effects on the uptake, intracellular distribution, and release of norepinephrine in the rat adrenal medulla. Results showed that 100 nM atrial natriuretic factor induced a rapid increase of norepinephrine uptake during the first minute of the incubation period. This increase was maintained for up to 60 min. In addition, only neuronal norepinephrine uptake was increased by the natriuretic factor; non-neuronal norepinephrine uptake was unaltered. Atrial natriuretic factor modified the intracellular distribution of the amine store: the granular fraction of norepinephrine increased, while the cytosolic fraction decreased. On the other hand, different concentrations (10, 50, and 100 nM) of the atrial factor decreased spontaneous [3H]norepinephrine output in a concentration-dependent manner. Furthermore, atrial natriuretic factor (10 nM) also reduced high potassium solution evoked secretion of norepinephrine. These results suggest that atrial natriuretic factor modulates sympathetic activity in the rat adrenal medulla. These effects of atrial natriuretic factor may be related to the catecholamine peripheral mechanism involved in the regulation of arterial blood pressure, smooth muscle tone, metabolic activity, etc. PMID- 8402383 TI - Catabolism of intracerebroventricularly injected 5-hydroxytryptamine in mouse: effect of coinjection of tryptamine and several pretreatments. AB - The catabolism of intracerebroventricularly injected 5-hydroxytryptamine in mouse brain was investigated. Pretreatment of animals with the 5-hydroxytryptamine type 1 receptor antagonist metergoline, the 5-hydroxytryptamine type 2 receptor antagonist ketanserin, the 5-hydroxytryptamine reuptake inhibitor fluoxetine, or the selective 5-hydroxytryptamine neurotoxin 5,7-dihydroxytryptamine failed to alter the rate of catabolism of intracerebroventricularly administered 5 hydroxytryptamine. The monoamine oxidase inhibitor tranylcypromine effectively blocked degradation of injected 5-hydroxytryptamine and accumulation of 5 hydroxyindoleacetic acid. Coinjection of tryptamine with 5-hydroxytryptamine reduced the rate of conversion of 5-hydroxytryptamine to 5-hydroxyindoleacetic acid. These results indicate that intracerebroventricularly administered 5 hydroxytryptamine is removed by a monoamine oxidase dependent system. This catabolism is not affected by inhibition of presynaptic uptake, 5 hydroxytryptamine receptor type 1 or type 2 blockade, or destruction of serotonergic nerve terminals. The coadministration of tryptamine may prolong the residence period of 5-hydroxytryptamine through competition for monoamine oxidase. PMID- 8402384 TI - Fever alters osmosensitivity of hypothalamic-vasopressin system in the rat. AB - The osmosensitivity of peripheral vasopressin release was studied during healthy thermoregulation and endotoxin-induced fever. There was an increase in osmosensitivity following bolus injection of saline in febrile rats. These animals displayed a steeper slope in the linear relationship between plasma osmolality and plasma vasopressin levels compared with afebrile animals. The change in regression slope was due to a significantly lower plasma osmolality in febrile rats. The plasma osmolality of animals infused with hypertonic saline was similarly decreased, but data analysis failed to show a significant change in the regression slope. Osmotic thresholds were not altered in either group. There was an increased urine output in febrile rats, and consequently these animals excreted greater amounts of salts than afebrile rats. This could account for the lower plasma osmolality observed in the febrile rat. PMID- 8402385 TI - Newborn piglet lungs release endothelin-1: effect of alpha-thrombin and hypoxia. AB - Endothelin-1 (ET-1) is a 21 amino acid vasoconstrictor peptide produced by endothelial cells, the expression of which is modulated by a variety of vasoconstrictors, vasodilators, and inflammatory mediators. Hypoxia has been shown to increase ET-1 expression and release in cultured endothelial cells from the systemic circulation, but reports are contradictory regarding the pulmonary circulation. In this study, the release of ET-1 and its cellular localization in the isolated perfused newborn piglet lung were examined under control conditions and after stimulation with hypoxia or alpha-thrombin (positive control). In the control condition, perfusion pressure remained stable during the study period, and a progressive increase in levels of immunoreactive ET-1 (irET-1) was noted. When alpha-thrombin was added to the perfusion fluid, a slow gradual increase in perfusion pressure was produced and the levels of irET-1 were significantly greater than those measured in the control preparations. Finally, hypoxia produced a significant increase in the perfusion pressure; however, the release of irET-1 did not differ significantly from the control, if anything, the net release across the lung was diminished. In all conditions, immunocytochemistry using antiserum to human-porcine ET-1 revealed the presence of high ET-1-like immunoreactivity in epithelial cells of bronchi, bronchioles, and terminal bronchioles. In addition, endothelial cells of large and medium-size pulmonary arteries were only moderately immunoreactive for ET-1. These findings indicate that the neonatal pig lung can produce and release ET-1, and that its release can be increased by certain stimuli like alpha-thrombin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402386 TI - Site specificity of fat cell lipolysis during pregnancy in two strains of rats. AB - The response to energy intake and expenditure is thought to be influenced by the genetic background. In the present study the metabolic response to pregnancy and to exercise training during pregnancy was investigated in two strains of rats. Lean Zucker and Wistar rats were divided into three groups: control pregnant (CP), exercise-trained pregnant (TRP), and not trained and not pregnant control (CNP). Trained rats swam 3 h per day, 6 days per week, throughout pregnancy. Body weight and food intake increased similarly during pregnancy in both strains (p < or = 0.05). However, only Wistar rats had a further increase of food intake and body weight during the second half of pregnancy: TRP weighed 29.1 more grams and ate 4.5 more grams of food per day than CP at the end of pregnancy (p < or = 0.05). Inguinal and parametrial fat cell sizes were unchanged during pregnancy. In both strains training induced a decrease of inguinal fat cell size at the beginning of pregnancy (p < or = 0.05), which was rapidly counteracted to reach CP values on day 20 of pregnancy. Parametrial fat cell size was also decreased by training (p < or = 0.05), but no values returned to control levels during pregnancy. In both strains, pregnancy increased fat cell lipolysis in the inguinal depot only (p < or = 0.05). Training during pregnancy inhibited fat cell lipolysis in inguinal and parametrial depots, especially in Zucker (p < or = 0.05), TRP reaching values similar to control values on day 20 of pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402387 TI - Active skeletal muscle metabolism and tension production: the influence of biopsies. AB - The influence of repeated sampling by the biopsy technique on skeletal muscle's metabolic and force-output responses was studied using the in situ canine gastrocnemius preparation. The left muscle was stimulated (8 V, 0.2 ms) for 1 h at 3 Hz. In the biopsy series (n = 9) muscle samples were taken at rest, and at 0.5, 2, 5, 15, 30, 45, and 60 min of stimulation. In the control series (n = 8) the left and right muscles were quick-frozen in N2 immediately after the 60 min of stimulation. The two series were not different in blood flow, VO2, arterial or venous [H+], muscle glycogen, or lactate release throughout the 60 min of activity. The lactate release was transient and was associated with an accumulation of intramuscular lactate and a period of rapid glycogenolysis. The biopsy series had a modest but significantly (p < 0.05) higher muscle lactate concentration both at rest and at the end of the contractions. The biopsy series also had less (p < 0.05) tension development throughout the hour; however, the O2 cost per unit of tension development was not different between groups, nor was the rate of tension decline over time different. This together with the similarities in perfusion, carbohydrate use, and lactate metabolism suggests that repeated biopsies had minimal impact on the muscle. The technique allows the collection of data over time; this improves the detail of experiments and means that fewer animals are required for a study. PMID- 8402388 TI - Steroidogenic enzyme activities in rat polycystic ovaries. AB - Ovaries containing multiple follicular cysts occur in a variety of anovulatory conditions. A macrocystic condition occurs spontaneously in rats following a single injection of estradiol valerate. The ovaries are small, and exhibit scant stromal tissue, few healthy follicles, and numerous large cystic and precystic follicles. We have also generated a microcystic condition by means of subcutaneous estradiol-containing silastic implants. These ovaries are large, and exhibit a stroma of hypertrophied lipid-filled cells, and numerous small cysts encircled by hypertrophied thecal cells. The macrocystic condition is associated with a uniformly attenuated plasma luteinizing hormone (LH) pattern, whereas large LH episodes characterize the microcystic condition. The marked dissimilarities between these two methods suggest that there may be corresponding differences in ovarian steroidogenic activity. We have measured the activity of enzymes involved in progestin and androgen biosynthesis in the two types of multicystic ovaries before and after LH - human chorionic gonadotropin (hCG) stimulation. Control ovaries were obtained at late proestrus from age-matched cycling animals. Radiometric enzyme assays for 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 20 alpha-hydroxysteroid dehydrogenase (20 alpha-HSD), C17,20-lyase (lyase), and aromatase were conducted on the microsomal fraction of ovarian homogenates. 3 beta-HSD activity was reduced by > 50% in both types of cystic ovaries compared with controls. There was a slight elevation in the 3 beta HSD activity of macrocystic ovaries in response to hCG. 20 alpha-HSD activity was similar in controls and macrocystic ovaries but significantly lower (< 20% of control) in the microcystic ovaries. Lyase and aromatase activities were undetectable in cystic ovaries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402389 TI - Modification of excitation-contraction coupling in cat ventricular myocardium following endocardial damage. AB - Damage to endocardial endothelium (denudation of the superficial tissue) by brief exposure to a 100-microL bolus of detergent (Triton X-100, 1% by volume stock) decreased the twitch force of papillary muscle (and trabeculae) by approximately 30% to a new but steady level without changes in resting tension. The decline in twitch force was evident immediately after the addition of Triton. Modification of the action potential measured from the contracting tissue appeared only later, when the change in contraction was already well established (i.e., after approximately 2 min). Action potential shortened in duration at 50% repolarization by approximately 100 ms and increased in plateau amplitude, although the latter increase was not always observed. A similar treatment procedure applied to strips of ventricular wall with the endocardium exposed to the superfusion solution resulted in a substantial decrease in action potential duration (approximately 110 ms). In contrast, treatment of strips of epicardial layers of ventricular walls (with epicardial side facing the superfusion solution) did not produce a similar result. In beta-stimulated (1 microM isoproterenol) and partially depolarized preparations (with 20 mM KCl), with intact endocardium, electrically evoked contractions were followed by aftercontractions, which were suppressed following Triton treatment. Action potentials in a depolarizing medium also shortened in duration (approximately 50 ms), although following a delay (2-3 min). The decay to steady state of postextrasystolic potentiated beat was slower after endocardial damage than under control conditions. This suggests an increased Ca2+ recirculation through the sarcoplasmic reticulum between two consecutive beats (35% before Triton vs. 45% after Triton).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402390 TI - Glucose-lowering effects of a new organic vanadium complex, bis(maltolato)oxovanadium(IV). AB - Inorganic vanadium has been shown, both in vivo and in vitro, to have insulin mimetic properties. A new organic vanadium complex, bis(maltolato)oxovanadium(IV) (BMOV), was developed to increase the absorption of vanadium from the gastrointestinal tract, thereby reducing the dose of vanadium necessary to produce glucose-lowering effects. BMOV was administered in the drinking water for 25 weeks to control and streptozotocin-induced diabetic, male Wistar rats. BMOV treatment produced a stable euglycemic state in 70% of diabetic treated animals. The other 30% of the diabetic treated animals demonstrated fluctuations in glucose control over the entire study period. The initial effective dose of BMOV was 0.45 mmol/kg, which decreased to an effective maintenance dose of 0.18 mmol/kg, significantly lower than the dose of inorganic vanadium salts used in previous studies. BMOV treatment did significantly reduce fluid consumption levels in control treated animals after 10 weeks of therapy; however, the food consumption for control treated animals was only intermittently lower than that for controls. Plasma cholesterol and triglyceride levels were normalized with BMOV treatment for all diabetic treated animals, without a concomitant increase in plasma insulin levels. An oral glucose tolerance test demonstrated that glucose homeostasis in control-treated animals occurred at significantly lower plasma insulin levels than in control animals. BMOV effectively produced the glucose-lowering effects at significantly lower dose than previously used for inorganic vanadium salts, without any overt signs of toxicity. PMID- 8402391 TI - Improvement in cardiac dysfunction in streptozotocin-induced diabetic rats following chronic oral administration of bis(maltolato)oxovanadium(IV). AB - Decreased cardiac function in streptozotocin-diabetic rats has been used as a model of diabetes-induced cardiomyopathy, which is a secondary complication in diabetic patients. The present study was designed to evaluate the therapeutic effect of a new organic vanadium complex, bis(maltolato)oxovanadium(IV), (BMOV), in improving heart function in streptozotocin-diabetic rats. There were four groups of male, Wistar rats: control (C), control treated (CT), diabetic (D), and diabetic treated (DT). Treatment consisted of BMOV, 0.5 mg/mL (1.8 mM) for the first 3 weeks and 0.75 mg/mL (2.4 mM) for the next 22 weeks, in the drinking water of rats allowed ad libitum access to food and water. BMOV lowered blood glucose to < 9 mM in 70% of DT animals without any increase in plasma insulin levels, and mean blood glucose and plasma lipid levels were significantly lower in DT vs. D rats. Tissue vanadium levels were measured in plasma, bone, kidney, liver, muscle, and fat of BMOV-treated rats. Plasma vanadium levels averaged 0.84 +/- 0.07 microgram/mL (16.8 microM) in CT rats and 0.76 +/- 0.05 microgram/mL (15.2 microM) in DT animals. The highest vanadium levels at termination of this chronic feeding study were in bone, 18.3 +/- 3.0 micrograms/g (0.37 mumol/g) in CT and 26.4 +/- 2.6 micrograms/g (0.53 mumol/g) in DT rats, with intermediate levels in kidney and liver, and low, but detectable levels in muscle and fat. There were no deaths in either the CT or DT group, and no overt signs of vanadium toxicity were present. Tissue vanadium levels were not correlated with the glucose-lowering effect.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402392 TI - Effect of carbonic anhydrase III inhibition on substrate utilization and fatigue in rat soleus. AB - Carbonic anhydrase III (CA III; EC 4.2.1.1) is the most abundant cytosolic enzyme in type I skeletal muscle fibers. We have previously shown that inhibiting the CA III activity of type I muscle can influence fatigability. Our goal was to test the hypothesis that the influence on fatigability of CA III inhibition is linked to an increased utilization of carbohydrates. Rat soleus muscles were incubated in vitro in a physiological solution with or without CA inhibitor (methazolamide, 1 mM) and submitted to a fatigue protocol. When the bathing solution contained glucose, the muscles incubated with methazolamide maintained a higher level of tension production than control muscles for the first 55-60 min of the test compared with 35-40 min when glucose was not added. Measurement of muscle glycogen content revealed that muscles incubated with CA inhibitor were utilizing their glycogen at a higher rate than control muscles over the first 45 min of the fatigue protocol. When glycolysis was inhibited with sodium iodoacetate, fatigability was not influenced by the addition of a CA inhibitor. These results further support the existence of a link between CA III activity and energy metabolism in type I skeletal muscle fibers. PMID- 8402393 TI - Chloride and depolarization by acetylcholine in canine airway smooth muscle. AB - The role of chloride channels has been examined in canine tracheal smooth muscle by recording mechanical responses to field stimulation and to acetylcholine (ACh) and by sucrose gap recording of excitatory junction potentials and ACh-induced electrical changes. The results of substitution studies using isethionate for chloride provided evidence that a chloride conductance contributes to the resting potential. The extrapolated reversal potential for ACh-induced depolarization was positive to the resting potential. Isethionate substitution inhibited ACh-induced depolarization, consistent with a contribution from increased Cl- conductance to the depolarization induced by ACh. However, closure of K+ channels and opening of a non-specific cation channel could also contribute to depolarization. Further study of the effects of isethionate substitution during prolonged tissue exposure to chloride-free medium showed that retention or the accumulation of Ca2+ in intracellular stores was impaired. We conclude that effects of chloride deprivation on responses to ACh may reflect an early increase in Cl- conductance, but longer term changes reflect the requirement for this anion to maintain internal Ca2+ stores. PMID- 8402394 TI - Hypometabolic and hypothermic factors from small intestine of hibernating ground squirrels (Citellus undulatus). AB - Low molecular mass components of the acetic acid extract from the small intestine of hibernating ground squirrels (Citellus undulatus) produced a decrease in oxygen consumption and body temperature of white mice and a dose-dependent delay in embryonic development of sea urchin (Strongylocentrotus intermedius). Equivalent doses of low molecular mass components obtained by the same method from active (summer) animals did not have such an effect. PMID- 8402395 TI - Impairment of endothelium-dependent relaxation of superior mesenteric artery in genetically diabetic WBN/Kob rats. AB - The endothelium-dependent relaxation of superior mesenteric arteries of Wistar and genetically diabetic WBN/Kob rats was compared. Endothelium-dependent relaxation induced by acetylcholine (ACh) and A23187 was depressed in WBN/Kob rats. Relaxation induced by sodium nitroprusside, an endothelium-independent agent, in strips from WBN/Kob rats was similar to that in strips from Wistar rats. Indomethacin (5 x 10(-6) M) enhanced the relaxation responses to ACh in strips from both WBN/Kob and Wistar rats; however, endothelium-dependent relaxation induced by ACh remained attenuated in WBN/Kob rats. These results show that endothelium-dependent relaxation is impaired not only in thoracic aorta but also in superior mesenteric arteries in genetically diabetic rats. PMID- 8402396 TI - Inhibition by nifedipine of endothelin-induced adrenal catecholamine secretion in anesthetized dogs. AB - The present study was conducted to investigate local effects of endothelin-1 (ET 1) on basal adrenal catecholamine (CA) secretion and underlying mechanisms of action of ET-1 in anesthetized dogs. ET-1 was locally administered to the left adrenal gland through a catheter inserted into the left adrenolumbar artery. Plasma CA concentrations in adrenal venous and aortic blood were determined by an HPLC-electrochemical method. The local infusions (1 min, 0.5 mL/min) of ET-1, in doses of 0.01, 0.1, and 1.0 microgram/mL, resulted in a dose-dependent increase in epinephrine secretion, rising from an initial basal value of 6.9 +/- 1.4 ng.min-1 x g-1 to maximum values of 7.6 +/- 1.5, 81.1 +/- 38.3*, and 680.9 +/- 107.6* ng.min-1 x g-1 (*p < 0.05, n = 7), respectively. Neither mean arterial pressure nor plasma CA levels in aortic blood altered following ET-1 infusions. A similar dose-dependent increase in norepinephrine output was also observed with ET-1. Following the initial peak observed during infusion with the highest dose of ET-1, the output of both epinephrine and norepinephrine remained significantly elevated over a period of 30 min. This ET-1-induced, long-lasting increase in adrenal CA output was abolished by nifedipine (100 micrograms/mL) similarly administered 10 min before ET-1 infusion. The results indicate that ET-1 directly enhances the basal secretion of CA from dog adrenal medulla in vivo. The study suggests that the ET-1-induced increase in adrenal CA secretion is, at least in part, mediated by the opening of dihydropyridine-sensitive L-type calcium channels at the level of adrenal medullary chromaffin cells. PMID- 8402397 TI - A dual approach to self-stimulation and locomotor trace affected by chronic methamphetamine treatment for an animal model of schizophrenia. AB - The effect of long-term methamphetamine (MAP) treatment on intracranial self stimulation of the lateral hypotholamus and locomotor traces was assessed. An attempt was made to provide a useful animal model for understanding anhedonia, stereotypy, and reoccurrence of liability, which are analogous to symptoms of schizophrenia. The frequency of intracranial self-stimulation (ICSS) as used as a measure of the animals' "hedonic-anhedonic" state. Following long-term MAP treatment (3 mg/kg), rats gradually showed stereotyped behavior, and became inactive and unresponsive to ICSS. These behavioral changes and decreased ICSS lasted several weeks after cessation of chronic MAP treatment and seemed to suggest post-MAP chronic psychosis and (or) anhedonia, two of the negative symptoms of schizophrenia. The traces of rat behavior affected by chronic MAP treatment were classified into three types, peripheral, mixed, and fixed, occurring in a dose-dependent manner. Reverse tolerance, similar to the reoccurrence of schizophrenic symptoms, was observed as a fixed stereotypy associated with loss of ICSS. These abnormal phenomena were suppressed by pretreatment with haloperidol. In the present study, the combination of ICSS and locomotor trace affected by chronic MAP treatment was proposed as an animal model of schizophrenia and as a useful technique for gauging the effect of neuroleptics. PMID- 8402398 TI - Lack of norepinephrine--neuropeptide Y interactions in the hypothalamic control of cardiovascular regulation in urethane-anesthetized rats. AB - Noradrenergic input to the hypothalamus is implicated in cardiovascular and behavioral regulation. The hypothalamus also contains high concentrations of neuropeptide Y (NPY), which often is colocalized in noradrenergic neurons. Systemically, NPY has been demonstrated to act synergistically with norepinephrine. Injections of concentrated solutions of NPY into the cerebral ventricles or hypothalamus have been found to alter ingestive behaviours, but the role of NPY in cardiovascular regulation is unknown. The objectives of this study were to determine if NPY injected directly into the hypothalamus elicits cardiovascular responses and (or) if the simultaneous administration of NPY with norepinephrine alters the cardiovascular responses elicited by norepinephrine alone. The hypothalamus of the urethane-anesthetized rat was mapped for heart rate and blood pressure responses to injections of one of the following: saline; NPY (1.9 or 0.6 pmol per injection site); norepinephrine (24, 81, or 243 nmol per injection site); or the combination of norepinephrine plus NPY. Site and drug selection were randomized. Injections of 0.25 microL were at 1.0 microL/min, bilaterally for bilateral structures and unilaterally for midline structures. Norepinephrine routinely elicited dose-dependent increases in blood pressure with latencies of approximately 1 min, which peaked by 3-5 min, accompanied or followed by tachycardia. Saline and NPY injections alone elicited no significant responses in any site. When NPY was injected together with norepinephrine, there were no significant alterations in cardiovascular variables except for attenuation of pressor responses when NPY was injected into the preoptic region and attenuation of tachycardia when NPY was injected into the caudal hypothalamus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402399 TI - Use of high and low responders to novelty in rat studies on the role of the ventral striatum in radial maze performance: effects of intra-accumbens injections of sulpiride. AB - High and low responders to novelty (Wistar rats) were selected with the help of an open-field test and then equipped with intra-accumbens cannulae. They were then tested in a simple four-arm radial maze during 5 successive days, three trials per day, following intra-accumbens injections of distilled water or the dopaminergic D2 antagonist (+/-)-sulpiride. The injections were given 15 min before the first trial on each day. Both types of drug-naive rats reached the same level of performance on day 5. However, high responders made more visits, more revisits, and needed less time to make the first visit than low responders. Moreover, high responders showed their greatest increase in learning 2 days earlier than low responders. It is discussed that these differences between high and low responders are not due simply to differences in locomotor activity, but are due to a subtle, but important, difference in the mode of learning between both types. Sulpiride significantly attenuated the learning in both rat types; however, its effect in high responders was much less than that in low responders. It is suggested that the effects of sulpiride are not due to changes in locomotor activity, motivation, or perception, but are due to a learning deficit. The data are discussed in view of the genetic variation in the neurochemical and neurobiological makeup of the nucleus accumbens in both types. PMID- 8402400 TI - Effects of medial prefrontal cortex stimulation on the spontaneous activity of the ventral pallidal neurons in the rat. AB - The neural connections from the medial prefrontal cortex to the ventral pallidum were investigated in urethane-anesthetized rats. Extracellular recordings were made from 124 spontaneously firing neurons in the ventral pallidum while the medial prefrontal cortex was electrically stimulated. The most frequent response to prefrontal cortex stimulation was inhibition of the firing rate of 72.3% of the neurons with orthodromic response (mean latency: 14.4 +/- 1.6 ms). Excitatory responses were found in 27.7% of the neurons with orthodromic response (mean latency: 8.5 +/- 1.4 ms). Frequency histograms of latencies were unimodal in both types of responses. Fifty-nine neurons (47.6% of the total tested) showed no change in spontaneous firing after medial prefrontal cortex stimulation. The electrophysiological results support previous anatomical findings of connections between the medial prefrontal cortex and the ventral pallidum. These projections play a predominantly inhibitory role in the spontaneous activity of ventropallidal neurons, and show topographical organization. This inhibition may modulate the motor performance of motivated behaviors. PMID- 8402401 TI - Cholinergic effects on spatial exploration and its memory. AB - In their first swim in an unfamiliar circular swimming pool, control rats showed declines in average swimming speed and in the time spent in the perimeter of the pool. Both declines were antagonized by the muscarinic antagonist scopolamine, but not by methylscopolamine, a muscarinic antagonist that crosses the blood brain barrier only poorly, indicating that these declines depend upon central cholinergic activity. In the first minute of a second swim 3 days later, control rats spent a much longer time in the central region of the pool than in the first minute of the first swim. This modification of behaviour by previous experience suggests that a long-term memory of the first swim was formed. Scopolamine, but not methylscopolamine, administered before the first swim attenuated this modification of behaviour. Pilocarpine, administered shortly after scopolamine before the first swim, significantly normalized all the scopolamine-induced changes, whereas oxotremorine and arecoline normalized only habituation of perimeter preference; agonists administered alone decreased swimming speed and perimeter preference without affecting their rates of decline. The results suggest that in this test, different cholinergic mechanisms are involved in habituation of swimming speed and habituation of perimeter preference. PMID- 8402402 TI - Convergence of ventrolateral medullary and aortic baroreceptor inputs in nucleus of the solitary tract. AB - Experiments were done in alpha-chloralose anesthetized rats to investigate the effect of stimulating the intermediate region of the ventrolateral medulla (VLM) on the response of single units in nucleus of the solitary tract (NTS) that altered their rate of discharge during aortic baroreceptor stimulation. Of 178 units recorded within NTS, 65 responded orthodromically to VLM stimulation. An additional 16 units were activated antidromically by VLM stimulation. The interaction between the aortic depressor nerve and VLM orthodromic inputs was investigated in 38 units that received converging inputs from VLM and the aortic depressor nerve. A conditioning stimulus applied to VLM, regardless of whether the NTS unit was excited by (n = 13) or did not respond to (n = 9) VLM stimulation, decreased the excitatory response of the unit to aortic depressor nerve stimulation. These data suggest that VLM neurons are involved in the modulation of aortic baroreceptor afferent information at the level of the NTS and that this interaction of inputs in NTS may involve both postsynaptic and presynaptic mechanisms. PMID- 8402403 TI - Response characteristics of amygdaloid neurons provoked by emotionally significant environmental stimuli in cats, with special reference to response durations. AB - Extracellular single or multiple neuronal activities were recorded from the basolateral portion of the amygdala of wild adult cats under an unanesthetized, freely moving condition, and neuronal responsiveness to neutral, aversive, and appetitive stimuli was studied. Of 71 units, 47 (66%) responded to some of the stimuli. The patterns of neuronal responses were classified into three types on the basis of response duration. Of the responses sampled, 9% rapidly attenuated and disappeared before termination of stimulus presentation (pattern A), 58% of responses were maintained during the period of the stimulus presentation but disappeared abruptly after termination (pattern B), and 33% of responses markedly outlasted the stimulus presentation period (pattern C). Pattern A responses habituated readily and were most prominent when neutral stimuli were presented, so this type of response was considered to underlie altering or orienting responses. Pattern B responses were observed equally for the three kinds of stimuli, and were suggested to be predominantly involved in perception of the environmental stimuli. Pattern C responses habituated least and tended to be elicited more frequently by aversive stimuli. This type of response was interpreted to reflect emotional arousal. These findings were considered to be compatible with the hypothesis that the amygdala plays an important role in converting environmental stimuli into emotions such as rage or fear. PMID- 8402404 TI - Water and solute balance in rats during 10 h water deprivation and rehydration. AB - Rats with bladder and venous cannulas were deprived of water from midnight (00:00) to 10:00. Water deprivation reduced food intake within 2 h, reducing the amount of water sequestered in the gut and the solute load to the tissues. There was little change in either urinary water loss or osmolality, but water-deprived rats excreted more Na+, K+, and Cl- than food-matched controls. The change in solute balance helped preserve osmolality and cell volume at the expense of extracellular fluid volume. When water was returned, rats quickly drank enough to restore the intracellular but not the extracellular fluid deficit. Plasma osmolality and sodium concentration fell below predeprivation values. Urine osmolality and excretion of Na+, K+, and Cl- fell rapidly after drinking. Drinking continued at a slower rate for at least 4 h, but urine flow also increased so water balance stabilized. The changes in intake and electrolyte excretion during water deprivation and rehydration illustrate the important role of changes in solute balance in fluid homeostasis. PMID- 8402405 TI - Increased extracellular dopamine in the nucleus accumbens of the rat elicited by a conditional stimulus for food: an electrochemical study. AB - Stearate-modified graphite paste electrodes were implanted chronically into dopamine terminal regions in the nucleus accumbens or caudate nucleus of the rat. Reverse dialysis was used to demonstrate a selective response of these electrodes to dopamine, but not 3,4-dihydroxyphenylacetic acid or ascorbic acid. In a separate behavioural experiment, a significant increase in the chronoamperometric response was observed during presentation of a conditional stimulus predictive of food, and the electrochemical response remained elevated during and following consumption of the meal. Similar trends were observed from electrodes in the caudate nucleus. These data confirm the activation of mesolimbic dopamine neurons by incentive stimuli predictive of food and possibly by consumption of food. Together with other recent data on sex- and thirst-related increases in dopamine levels in the nucleus accumbens, these findings are consistent with a role for the nucleus accumbens as an interface between motivation and activation of the motor system. PMID- 8402406 TI - Decreases in rat locomotor activity as a result of changes in synaptic transmission to neurons within the mesencephalic locomotor region. AB - The mesencephalic locomotor region is defined as a functional region sending signals to the spinal cord generators of rhythmical limb movements for locomotion. It has been shown that the mesencephalic locomotor region plays a critical role in locomotion initiated from the nucleus accumbens or from the subpallidal region. However, there are conflicting data on whether synaptic input from the nucleus accumbens--subpallidal region to the mesencephalic locomotor region mediates locomotion. The purpose of the study was to determine the role of synaptic input to different subregions of the mesencephalic locomotor region in locomotion induced by injecting dopamine into the nucleus accumbens or by injecting picrotoxin into the subpallidal region in freely behaving rats. Synaptic transmission in the mesencephalic locomotor region was eliminated by excitotoxic lesions or was reversibly interrupted by injecting cobalt chloride, which can block synaptic transmission. Excitotoxic lesions or injections of cobalt into subregions of the mesencephalic locomotor region significantly decreased, although did not completely block, locomotion. The most effective sites for cobalt- and lesion-induced reduction in locomotion were consistent with localization of the mesencephalic locomotor region. Effective sites for cobalt and lesions markedly overlapped but were not identical. The results indicate that synaptic transmission within the mesencephalic locomotor region contributes to dopamine- or picrotoxin-induced locomotion. PMID- 8402407 TI - Anatomical distribution of brainstem sites where PGE1 induces hyperthermia in macaque species. AB - The neuroanatomical distribution of sites in the diencephalon and mesencephalon within which a prostaglandin (PG) of the E series elicits hyperthermia was characterized in Macaca mulatta and Macaca nemestrina. In 420 experiments undertaken in 13 animals, 225 loci were examined for their reactivity to PGE1 microinjected in a dose of 30 or 100 ng given in a volume of 1.0-1.5 microL. The regions of the brainstem for injection extended rostrally from the thermosensitive cells of the anterior hypothalamic, preoptic area (AH/POA) to the caudal border of the mesencephalon. Colonic and skin temperatures of the monkeys were measured continuously by thermistor probes. A hyperthermic response of > or = 0.5 degrees C and a latency of < or = 45 min was evoked by PGE1 within sites located primarily in the AH/POA. When PGE1 was microinjected at loci located caudal to the AH/POA, the elevation in body temperature (Tb) not only was less intense but rose at a slower rate. A higher concentration of PGE1 in these caudal regions was required to induce hyperthermia comparable with that elicited at loci within the AH/POA. In a second series of experiments either 1.0-5.0 micrograms 5 hydroxytryptamine (serotonin) or a concentration of 10(8) organisms/mL of Escherichia coli was microinjected at PGE1-reactive sites. A close anatomical concordance within the AH/POA of the animal was found in terms of the temporal characteristics and magnitude of the hyperthermia evoked by the indoleamine or lipopolysaccharide. The present results coincide with the reported neuroanatomical distribution of sites in the diencephalon and mesencephalon of other species in which PGE1 causes hyperthermia. Furthermore, these findings support the concept that the local neuronal mechanism of action of a pyrogen in the brainstem of the primate may involve phasic changes in the endogenous activity of both the serotonergic pathway and cyclo-oxygenase system in the AH/POA. In turn, their commonality of action suggests a functional similarity in their effect of shifting the set point for Tb. PMID- 8402408 TI - Age and organ transplantation. PMID- 8402409 TI - General Council uses annual meeting to send message of support to young MDs. PMID- 8402410 TI - AIDS in the US: a decade of fear and cruel indifference. PMID- 8402411 TI - Scientific integrity. PMID- 8402412 TI - Scientific integrity. PMID- 8402413 TI - CMA Gender Issues Committee. PMID- 8402414 TI - CMA Gender Issues Committee. PMID- 8402415 TI - Canadian physicians and euthanasia. PMID- 8402416 TI - Canadian physicians and euthanasia. PMID- 8402417 TI - Moving to the US. PMID- 8402418 TI - Radiologic recommendations in cases of gastric ulcer. PMID- 8402419 TI - Legalization of all drugs. PMID- 8402420 TI - Legalization of all drugs. PMID- 8402421 TI - The true story about Saudis. PMID- 8402422 TI - Drug names and medication errors: who is responsible? PMID- 8402423 TI - Seroprevalence of and risk factors for HIV-1 infection in injection drug users in Montreal and Toronto: a collaborative study. AB - OBJECTIVE: To determine the prevalence of antibodies to HIV-1 and risk factors for HIV-1 infection among injection drug users. DESIGN: Questionnaire survey. A venous blood sample was taken for HIV-1 antibody testing. SETTING: Montreal and Toronto. PARTICIPANTS: A total of 810 subjects who had used injection drugs in the previous 6 months recruited mainly from treatment centres and from the street in Montreal (425 subjects) and from treatment centres in Toronto (385 subjects) between September 1988 and September 1990. The overall participation rate was 82%. OUTCOME MEASURES: HIV-1 seropositivity, sociodemographic and behavioural risk factors for HIV-1 infection. RESULTS: The overall seroprevalence rate of HIV 1 infection was 4.8% (95% confidence limits [CL] 3.5 and 6.5). In Montreal the rate was 8.2% (95% CL 6.0 and 11.2), and in Toronto 1.0% (95% CL 0.4 and 2.6) (p < 0.001). Seropositive subjects were significantly older (p = 0.041) and were more likely to have a history of imprisonment (p = 0.006) than seronegative subjects. In univariate analysis seropositivity was associated with the following behaviours: more frequent cocaine use (p < 0.001), injecting drugs in "shooting galleries" (p = 0.002), sharing equipment with a person known to be HIV-1 seropositive (p = 0.006), "booting" fresh blood (p = 0.004), homosexual or bisexual orientation (p = 0.006), engaging in prostitution (p < 0.001) and, for men, number of male sexual partners in the previous 6 months (p = 0.007). In multivariate analysis the determinants of HIV-1 seropositivity were Montreal as the city of recruitment (odds ratio [OR] 6.7, 95% CL 2.32 and 19.42), engaging in prostitution (OR 2.13, 95% CL 1.01 and 4.75), a history of imprisonment (OR 3.51, 95% CL 1.33 and 9.29) and sharing equipment with a person known to be HIV-1 seropositive (OR 4.43, 95% CL 1.43 and 13.74). CONCLUSIONS: Our findings show that HIV-1 is circulating among injection drug users in Montreal and Toronto and that both drug use and sexual behaviours are implicated in the transmission of infection in the populations studied. Adapted preventive programs should be developed to prevent further spread of HIV-1 infection in this population. PMID- 8402424 TI - Waiting for coronary revascularization in Toronto: 2 years' experience with a regional referral office. AB - OBJECTIVES: To determine the frequency of major adverse events among patients awaiting coronary revascularization; to assess the match between referring physicians' estimates of urgency, a computer-generated multifactorial urgency rating score and actual waiting times; to determine the changes in waiting times as capacity for bypass surgery increased; and to evaluate the influence of choice of procedure or operator on waiting times. DESIGN: Consecutive case series. SETTING: Greater Toronto region. SUBJECTS: All 571 patients referred to an organized referral office by cardiologists at hospitals without on-site revascularization facilities between Jan. 3, 1989, and June 30, 1991. MAIN OUTCOME MEASURES: Preoperative fatal or nonfatal myocardial infarction; proportions of patients waiting longer than the maximum period recommended for their urgency rating; mean waiting times for various subgroups; and correlations among referring physicians' urgency ratings, computer-generated multifactorial urgency scores and waiting times. RESULTS: Of the 496 patients accepted for a procedure 5 had fatal cardiac events and 3 nonfatal myocardial infarction. Events occurred three times more often in patients with left mainstem disease than in those in other anatomic categories (relative risk [RR] 3.05, 95% confidence interval [CI] 1.48 to 6.27, p = 0.03). Both the computer-generated scores and the referring physicians' scores were correlated with the actual waiting time (r = 0.46 and 0.57 respectively). Waiting times and the proportion of patients with excessive waiting times fell during the study period (p < 0.0001). However, urgent cases were much less likely to be done "on time" than those with a recommended waiting time of more than 2 weeks (RR 0.16, 95% CI 0.11 to 0.25, p < 0.0001). The mean wait for coronary artery bypass grafting (CABG) was 22.73 days if the referral office was allowed to find a surgeon or interventional cardiologist and 35.31 days if one was requested (p = 0.002 after adjustment for urgency scores). CONCLUSIONS: Death of a patient on the waiting list is uncommon in an organized referral system. Patients with left main-stem disease are at higher risk of death than those in other anatomic categories. There were significant correlations between referring physicians' ratings of urgency, multifactorial urgency scores and actual waiting times. Expansion of capacity for CABG led to shorter waiting times, but patients with unstable symptoms continued to wait longer than recommended. Requests for a specific surgeon caused significantly longer delays. PMID- 8402425 TI - Sex-related differences in coronary revascularization practices: the perspective from a Canadian queue management project. AB - OBJECTIVE: To assess sex-related differences in coronary revascularization practices in a Canadian setting. DESIGN: Prospective analytic cohort study. SETTING: Regional referral office in Toronto. PATIENTS: A selected but consecutive group of 131 women and 440 men referred by cardiologists for revascularization procedures between Jan. 3, 1989, and June 30, 1991. INTERVENTIONS: Coronary artery bypass grafting (CABG) or percutaneous transluminal coronary angioplasty (PTCA). Nurse-coordinators placed the referral with a surgeon or interventional cardiologist at one of three hospitals, who then communicated directly with the referring cardiologist. MAIN OUTCOME MEASURES: Symptom status at referral, procedures requested and performed, and time from referral to procedure. RESULTS: Although the women were more likely than the men to have unstable angina at the time of referral (odds ratio [OR] 2.28, 95% confidence interval [CI] 1.38 to 3.79, p = 0.0006), more women than men (16.8% v. 12.1%) were turned down for a procedure. Significant sex-related differences in practice patterns (p < 0.001) persisted after controlling for age or for the referring cardiologists' perception of expected procedural risk. A stepwise multivariate model showed that anatomy was the main determinant of case management; sex was the only other significant variable (p = 0.016). The referring physicians requested CABG more often for men than for women (p = 0.009), and the men accepted for a procedure were much more likely to undergo CABG than the women (OR 2.40, CI 1.47 to 3.93, p = 0.0002). Although the women undergoing CABG waited shorter periods than the men (p = 0.0035), this difference was attributable to their more severe symptoms. CONCLUSIONS: In this selected group women had more serious symptoms before referral but were turned down for revascularization more often than men. Reduced use of CABG rather than PTCA largely accounted for the sex-related differences in revascularization. Once accepted for a procedure women had shorter waiting times, which was appropriate given their more severe symptoms. PMID- 8402426 TI - Profile of Canadian physicians: results of the 1990 Physician Resource Questionnaire. AB - OBJECTIVE: To determine the supply, mix and distribution of physicians in Canada and to compare data with those of the 1982 and 1986 physician surveys. DESIGN: National census mail survey. SETTING: Canada. PARTICIPANTS: All physicians licensed to practise medicine in Canada, excluding interns and residents. A total of 52,422 questionnaires were mailed, of which 771 were ineligible. There were 38,313 valid responses (response rate 74.2%). MAIN OUTCOME MEASURES: Activity status, workload, specialty certification, practice setting and demographic profiles. MAIN RESULTS: A total of 88.7% of the respondents were active physicians; 19.4% were women, compared with 16.8% in 1986. Physicians reported working on average 4.1 fewer hours per week in total activities than in 1986 and 5.7 fewer hours per week than in 1982. As was found in 1982, about 50% of active physicians were certified specialists; 30% of specialists and 21% of general/family practitioners were 55 years of age or more. Approximately 11% of active physicians were in rural practice, as was reported in 1986. Similar proportions of foreign graduates and Canadian graduates were located in rural areas (10.9% and 11.4% respectively). CONCLUSIONS: Factors such as aging and retirement will affect specific specialty groups (e.g., general surgery and obstetrics/gynecology) in the near future. Specialty groups must address the issue of the future supply of physicians and the demand for their services when developing targeted needs within their specialties. The increasing proportion of women in medicine is changing the specialty mix and practice profiles of physicians as a whole. The issues associated with the recruitment and retention of physicians in rural areas remain complex. PMID- 8402427 TI - Connective tissue disease: closing on the threshold. PMID- 8402428 TI - The heart of the matter: Dalton Camp and his controversial transplant. PMID- 8402429 TI - Progress in child psychiatry. PMID- 8402430 TI - Family factors in adolescent unipolar depression. AB - This paper reviews the current literature on the contribution of family factors to unipolar depression among adolescents. Research which examined the following factors was reviewed and evaluated: genetic transmission, parental depression, parental death, quality of attachment and family interaction. Studies had to meet the following criteria to be included: publication must have been between the years 1985 and 1992; the evaluation had to be empirical with some form of control; a dependent measure of family characteristics or functioning; the adolescent had to be between the ages of 13 to 19; and the adolescent had to meet the recognizable criteria for unipolar depression. Recent literature suggests that adolescents suffering from depression share a number of risk factors: a family history of affective disorder, inept or inadequate parenting, abrasive interactions within the family and insecure attachments. At this time, there is no evidence that these aversive interactions predate the disorder. Weaknesses in the current literature are identified and strategies for improving future research are proposed. PMID- 8402431 TI - Child survivors of the Holocaust--strategies of adaptation. AB - Child survivors have only recently been recognized as a developmentally distinct group with psychological experiences different from older survivors. The wartime circumstances of Nazi persecution caused enforced separation from family and friends, and all the survivors experienced persecution in the form of physical and emotional abuse, starvation and degradation, and were witnesses to cruelty. This paper is based on information from interviews and therapy with 25 child survivors, the majority of whom were not patients. Coping strategies are discussed in terms of their survival value in wartime and post-war adaptive value. Three themes which reverberate throughout the lives of child survivors, now adults, are discussed in greater detail: bereavement, memory and intellect. The fact that the majority of child survivors live normal and creative lives provides an opportunity to learn what factors have served them over 40 years, to provide the resilience and strength to cope after such a shattering beginning. PMID- 8402432 TI - A psychiatric follow-up study of adoptees. AB - This is a five year follow-up study of adopted children and adolescents. Thirty five adoptees and 23 control subjects were assessed. Five years earlier, the initial sample consisted of 57 pairs of adoptees and controls. The study showed that both adopted and control subjects were improved at the follow-up assessment and that there were no significant differences in clinical diagnoses and social adaptation between the groups. Compared with the controls, the adoptees were scored higher on a behaviour scale (Revised Behavior Problem Checklist) by parents. Adoption by the sixth month of age was associated with better overall psychosocial functioning. Significantly more adoptees were not living with their adoptive families. Factors associated with outcome are discussed. PMID- 8402433 TI - Evaluation of the original Ontario Child Health Study scales. AB - This article presents evaluative information on the use of the original Ontario Child Health Study scales to serve as original-level measures of conduct disorder, hyperactivity and emotional disorder among children in the general (non clinic) population. Problem checklist assessments were obtained from parents and teachers of children aged six to 16 and youth aged 12 to 16 drawn from a general population (n = 1,751); and a mental health clinic sample (n = 1,027) in the same industrialized, urban setting. The results showed that the original OCHS scales possess adequate psychometric properties to be used as original-level measures of disorder. Correlations between individual items and their hypothesized scales were very strong, indicating convergent validity, while correlations between the same items and other (non hypothesized) scales were lower, indicating discriminant validity. Item analyses indicated that individual scale items possess both convergent and discriminant validity. Although the scales were skewed to the positive end of the continuum, they demonstrated good internal consistency (all estimates > or = 0.74) and test-retest (all estimates > or = 0.65) reliability. Finally, three different validity analyses confirmed hypotheses about how the original OCHS scales should perform if they provide useful measures of disorder. PMID- 8402434 TI - Informants, correlates and child disorders in a clinical population. AB - This paper examines associations between informants, correlates and child disorders in a sample of 230 outpatients six to 14 years of age. K-SADS-E diagnoses and scores were studied according to informant, gender, age group, mental health of the parents, life events and parents' relationship. The results show that parents underreport their daughters' internalized disorders compared with reports of children and adolescents and that male children and adolescents underreport all types of disorders compared with the reports of their parents. Intra-informant correlations and differences between informants suggest an uneven influence according to informants of correlates on the report of child disorders. PMID- 8402435 TI - Measuring self-perceived role competence among first nations and non native children. AB - The objective of this research is to develop and test measures of self-perceived academic and social competence among First Nations and non native children. The method used is the analysis of psychometric properties of scales derived from questionnaires administered to First Nations children four culture areas of North America, as well as comparison samples of non native children. The results consist of the reliability coefficients, which fall into a satisfactory range; an internal consistency which increases with age; an agreement between self- and teacher-rated competence which is higher for non native than for native children. In the second grade, the competence scores of the native and non native children were equal. Thereafter, the scores of the native children either declined or remained static, while the non native scores tended to increase. In conclusion, the Flower of Two Soils scales are suitable for children from elementary schools, from First Nations and the majority culture. As children mature, the assessments of competence become an increasingly stable part of their repertoire of self percepts; asynchronous socialization may adversely affect self-perceived competence. PMID- 8402436 TI - Attachment and conduct disorder: the Response Program. AB - An increasing number of youths are being identified as suffering from behavioural problems that cause difficulties in their family and peer relations which in turn reduces their chances of academic and vocational success. There is growing concern regarding their level of aggressiveness. The common diagnosis given to these disaffiliated youths is conduct disorder. To date, most treatment programs for conduct disorder have been unsuccessful. A review of recent studies indicates that the disruption of attachment may be an important feature that underlies the wide range of symptoms that are typically found in youths with conduct disorder. A community-oriented program designed to ensure long term care for these youths is described in this paper, and the findings of a six month follow-up evaluation are presented. Results indicated that communities, caregivers, and youths responded positively to the program; caregivers reported significant reductions in a broad range of psychiatric symptoms in youths, and youths reported a significant reduction in symptoms of conduct disorder. PMID- 8402437 TI - A comparison of day treatment and outpatient treatment for children with disruptive behaviour problems. AB - The authors explored the relative merits of outpatient and day treatment for 30 children with severe behaviour problems. The effectiveness of treatment on behaviour, self-perception, and social and family functioning was assessed. Day treatment was found to be more effective in reducing behaviour problems, alleviating depressive symptoms, increasing social skills and improving family functioning. PMID- 8402438 TI - Public health nurse home visitation for the tertiary prevention of child maltreatment: results of a pilot study. AB - The purpose of this pilot study was to assess the feasibility of using public health nurse home visitation for the tertiary prevention of child maltreatment. The intervention was designed to provide support to the parents, education about child-rearing and child development and to assist families obtaining help from other services. Eighteen families referred from the Hamilton-Wentworth child protective agencies participated in the intervention. Enrolment criteria included first episode of suspected or verified child physical abuse or neglect, ongoing involvement with a child protective agency, index child less than 12 years of age and remaining in the home, and agreement to participate in the study. Home visits were provided weekly on average over a period of six months and the nurses were available by telephone. Pilot assessment included review of medical records, semi structured interviews to gather background information, checklists of child rearing attitudes, child behaviour and development. The participants were interviewed about their response to the intervention. The high degree of mobility of the families posed a problem in recruitment. Nevertheless, the public health nurses were able to develop a working alliance with 14 of the 18 families (77%) who participated. The participants gave permission for review of their medical records and reported that the methods of assessment were acceptable. The authors conclude that the intervention and assessment procedure are feasible and acceptable for preventing the recurrence of child maltreatment. Any decision about effectiveness of the intervention awaits a longitudinal, controlled trial. PMID- 8402439 TI - Parent training and social skills training for children with attention-deficit hyperactivity disorder: how can they be combined for greater effectiveness? AB - Attention-deficit hyperactive disorder (ADHD) is a chronic and treatment refractory syndrome affecting academic, social and emotional adjustment in children. Stimulant medication is the treatment of choice and is often paired with psychosocial treatment. However, no single treatment modality alleviates the symptoms or improves the negative peer status of these children in their social ecology over the long term. This article reviews two psychosocial treatments used for ADHD, parent training and social skills training and suggests ways that these two components may be combined for greater effectiveness by encouraging the parent to: 1. learn more about the importance of developing social competence and positive peer status; 2. use incidental teaching and self-evaluation strategies; 3. become strategic organizers of the child's social life; and 4. become case managers to facilitate more consistency between the significant adults in the child's social environment. PMID- 8402440 TI - Aims and methodological problems in multimodal treatment studies. AB - The paper outlines the growing acceptance and use of multimodal treatment approaches to treat a variety of psychiatric disorders in childhood and adolescence. It then explores some of the problems of multimodal treatment studies. Problems in assessing outcome of efficacy of the interventions, particularly as they pertain to specific interventions and particular areas of functioning are discussed as well as the issues of appropriate control and comparison groups. The problems with regard to appropriate control and comparison groups are explored with respect to diagnosis and interventions. Finally, an ongoing multimodal treatment study of children with attention deficit hyperactive disorder is used to illustrate how some of the problems described can be solved. PMID- 8402441 TI - Electroconvulsive therapy. PMID- 8402442 TI - Annual screening using chest x-ray examination for the diagnosis of lung cancer. PMID- 8402443 TI - Fine-needle aspiration of parotid masses. AB - BACKGROUND: There is controversy concerning the utility of fine-needle aspiration in diagnosing parotid masses. Even studies on large series of patients have compared aspiration findings with the histology in much fewer cases. METHODS: Preoperative fine-needle aspiration findings were compared with the histopathologic diagnoses from surgically resected specimens in 246 patients presenting with and treated for parotid mass from 1980-1990. RESULTS: Of 173 benign tumors, 159 (91.9%) were diagnosed correctly and 110 of 144 (> 60%) were typed. Of 36 malignant tumors, malignancy was recognized in 22 cases (61.1%). There were nine false-negatives, and in five cases, the specimen was unsatisfactory. The four cases of metastatic disease were correctly typed. Only two of seven lymphomas (28.6%) were identified. The cytologic and histologic diagnoses were concordant in all cases of nonneoplastic disease. Overall accuracy was 87%. CONCLUSIONS: Fine-needle aspiration speeds up the diagnostic process and, with close cooperation between clinician and pathologist, the technique is a valuable adjunct to preoperative assessment in patients with parotid masses. PMID- 8402444 TI - Long-term results of transhiatal esophagectomy for esophageal carcinoma. A multivariate analysis of prognostic factors. AB - BACKGROUND: Perioperative mortality and survival after esophagectomy have improved over the past 10 years. Although stage is the most powerful predictor of long-term survival, it remains unclear whether other factors influence prognosis. METHODS: Between 1981-1991, 100 patients with esophageal carcinoma were uniformly treated by transhiatal esophagectomy without adjuvant therapy. Results and prognostic factors of long-term survival were analyzed by univariate and multivariate analyses (log-rank test and Cox regression model). RESULTS: Forty eight patients had severe associated medical conditions, and 26 patients were older than 69 years of age. Mortality was 3%, and morbidity was 68%. With a median follow-up of 52 months, median survival was 18 months. The overall 5-year survival was 23%, but it was 63% for early stages (pT1 + pT2). In the multivariate analysis, the risk of dying was increased by 4.9 (risk ratio) for patients with carcinomas invading beyond the muscularis propria (pT3 + pT4), compared to lower stages (pT1 + pT2) (P < 0.0001). To a lesser extent, longterm survival was also adversely affected by transfusions (packed erythrocytes) after controlling for stage (risk ratio 1.7; P = 0.047). Age (> 69 years), preoperative weight loss, tumor location, histology (adenocarcinoma versus squamous cell carcinoma), fresh frozen plasma, and splenectomy did not influence survival. CONCLUSION: In this study, transhiatal esophagectomy provided palliation for esophageal cancer with a low-perioperative mortality. Prolonged survival or cure was obtained for the majority of patients operated on in the early stages. Blood transfusions had a slight adverse effect on long-term survival. PMID- 8402445 TI - Five-year survival after transhiatal resection of esophageal carcinoid tumor with a lymph node metastasis. AB - BACKGROUND: Carcinoid tumors of the esophagus are rare. Most reported cases have had a poor prognosis. The authors report the case of a 64-year-old man with a 4 cm carcinoid tumor of the lower esophagus. METHODS: Following endoscopic resection of an argyrophilic, nonargentaffin, carcinoid tumor of the lower esophagus, endosonography showed residual tumor, suggestive of a metastatic lymph node. The findings were confirmed at transhiatal esophagectomy. RESULTS: No recurrence has been observed along the current 5-year follow-up. CONCLUSIONS: Lymph node metastasis does not rule out the possibility of prolonged survival in esophageal carcinoids. Endosonography is strongly advisable for preoperative evaluation. PMID- 8402446 TI - Atypical adenomatous hyperplasia of the lung and its differentiation from adenocarcinoma. Characterization of atypical cells by morphometry and multivariate cluster analysis. AB - BACKGROUND: Atypical adenomatous hyperplasia (AAH) of the human lung is considered to be an important lesion preceding adenocarcinoma, but its difference from well-differentiated adenocarcinomas is so subtle as to cause diagnostic uncertainty. In view of this, the authors undertook to establish reproducible microscopic criteria for AAH, Type II pneumocyte type, and Clara cell type adenocarcinomas. METHODS: Twelve-parameter morphometry of atypical cells was performed on 97 lesions selected from 303 surgical specimens of lung by routine microscopic examination: all were considered by premorphometry examination to correspond to one of the above three diseases. Measurements were performed on photomicrographs using a digital image analyzer. The data of morphometry were subjected to 12-variate cluster analysis using a mainframe computer. RESULTS: It was demonstrated that the lesions were classifiable into three groups: Cluster 1 (Type II cell adenocarcinoma and AAH), Cluster 2 (AAH), and Cluster 3 (Clara cell adenocarcinoma). Whereas the latter two were created as homogeneous clusters, Cluster 1 was a mixture of Type II tumors and AAH. CONCLUSIONS: AAH in the strict sense of the word is definable by the features of those classified into Cluster 2, with atypia milder than overt adenocarcinomas. These AAH are likely to correspond to one of the steps of carcinogenesis forgoing the final one. The lesions, diagnosed as AAH before morphometry and being assigned to Cluster 1 and therefore not separable from Type II carcinoma, are considered to have been Type II carcinoma from the very beginning, which were however underdiagnosed in routine microscopic examination. PMID- 8402447 TI - Limitation of annual screening chest radiography for the diagnosis of lung cancer. A retrospective study. AB - BACKGROUND: The incidence and mortality rates of lung cancer have been increasing in many countries. However, the true effectiveness of screening for lung cancer is still controversial. This study aimed to examine the growth pattern of lung cancer and to evaluate the efficacy of screening. METHODS: The authors linked the records of annual radiologic screening to cancer registry data and conducted a retrospective follow-up study of radiographs in all patients with lung cancer arising in a population undergoing screening. RESULTS: Among a total of 305,934 participants, screening detected 206 lung cancers, 103 of which were Stage I disease. Seventy-one of the 131 adenocarcinomas were Stage I, and 58% of them showed evidence of cancer for 2 years on a retrospective review of radiographs. Presentation as small faint lesions overlapping the normal chest structures delayed the early detection of adenocarcinoma. The overall sensitivity of screening was 70%, 52% for squamous cell carcinoma and 50% for small cell carcinoma. Rapidly growing Stage II-IV tumors without retrospective evidence of cancer on previous radiographs accounted for most of the cancers detected during the intervals between screening. CONCLUSIONS: Both the low detectability of Stage I adenocarcinoma and the late recognition of rapidly growing small cell and squamous cell carcinomas reduced the effectiveness of screening. More effective imaging methods and an antismoking campaign are required to reduce lung cancer mortality. PMID- 8402448 TI - Characterization of lung squamous cell carcinoma-derived T-cell suppressive factor. AB - BACKGROUND: The immunosuppressive state of a tumor-bearing patient is possibly mediated by tumor-derived factor. In this study, the authors characterized lung squamous cell carcinoma-derived immunosuppressive factor (LSCF). METHODS: The immunosuppressive activity of QG56 (a lung squamous carcinoma cell line)-derived LSCF was evaluated by the effect of culture supernatant of QG56 on anti-CD3 monoclonal antibody-induced T-cell, response such as proliferation (3H-thymidine uptake), cytotoxicity (51Cr-releasing assay), and expression of cytokine mRNA (polymerase chain reaction). The LSCF was partially purified with an ion-exchange high-performance liquid chromatography (HPLC) and a gel-filtration HPLC. RESULTS: The LSCF inhibited proliferation, cytotoxicity, and expression of cytokine mRNA of T-cells in a dose-dependent manner. It has a molecular weight of approximately 22 kd, and was sensitive to proteinase K, heating at 60 degrees C, and resistant to treatment with trypsin and pH 3 and 9. These properties appear to be similar to those of transforming growth factor-beta (TGF-beta). However, the activity of the LSCF was not abrogated by anti-TGF-beta sera, and the LSCF did not suppress the proliferation of TGF-beta-sensitive mink lung cells (Mv1Lu). CONCLUSIONS: These data suggest that LSCF may be a novel tumor-derived immunosuppressive protein factor. PMID- 8402449 TI - Mediastinal paragangliomas. A clinicopathologic and immunohistochemical study of 16 cases. AB - BACKGROUND: Paragangliomas of the mediastinum are rare neoplasms. Because of their rarity, the spectrum of morphologic and immunohistochemical features and biologic behavior of these tumors in this location has not been delineated. METHODS: A retrospective study of 16 cases of mediastinal paragangliomas was undertaken to evaluate their clinicopathologic and immunohistochemical features. RESULTS: The patients were 10 men and 6 women whose ages ranged from 16-69 years of age (mean, 42.5 years). Twelve tumors were located in the posterior mediastinum, and three tumors were located in the anterior mediastinum. In one case, the exact location of the tumor within the mediastinum was not available. Microscopically, the nesting pattern (zellballen) was the most commonly encountered. Areas of stromal hyalinization were also frequent and were marked in two tumors. In two cases there was a prominent spindle cell component, and two cases showed granular cell changes in the cytoplasm of the tumor cells. Cellular pleomorphism was observed in the majority of cases. Occasional mitoses were seen in a few cases. Immunohistochemical studies in 10 cases showed chromogranin reactivity in 10 of 10 cases, S-100 protein in 9 of 10 cases, leu-enkephalin in 6 of 10 cases, and neurofilament protein in 5 of 10 cases. Keratin was negative in all cases studied. All patients underwent surgical resection of their tumors, and one patient underwent adjuvant radiation therapy. Follow-up information ranging from 1-168 months (mean, 84.5 months) was obtained in 12 patients. Nine patients (75%) were alive and well at last follow-up. In one patient the tumor metastasized to spinal space and bone marrow, and another patient died with metastases to lymph node, lung, and pelvis 14 years after initial diagnosis. CONCLUSIONS: The only parameter found to correlate with aggressive behavior was the extent of circumscription and/or local infiltration of the tumor at initial resection. No clinical, histologic, or immunohistochemical features were otherwise found to separate aggressive from indolent clinical behavior. Regular follow-up is recommended in these patients due to the potential for these tumors to metastasize. PMID- 8402450 TI - Vitiligo autoantibodies are effective against melanoma. AB - BACKGROUND: Vitiligo is a dermatologic disease characterized by local, dispersed, or diffuse white patches on the skin. The disease is defined as an autoimmune disorder because autoantibodies against membranal components of melanocytes are found in the patients' sera. The current study examined whether the autoantibodies reacting with the normal melanocytes could be a potent therapy against melanoma cells. METHODS: The three in vitro assays used to determine the antibody reactivities using a mouse melanoma cell line B-16-F10 and M-14 human melanoma cells as targets are as follows: enzyme-linked immunosorbent assay (ELISA), proliferation assay, and morphologic examination in the presence of antibodies purified from sera of patients with vitiligo. In the in vivo studies, experimental melanoma was intravenously induced in C57BL/6J mice, and the mice were treated by daily intraperitoneal injections with purified immunoglobulin G (IgG) fraction derived either from patients with vitiligo or from healthy subjects. RESULTS: The binding of IgG derived from patients with vitiligo was demonstrated by ELISA: Exposure of melanoma cells to the vitiligo autoantibodies was followed by inhibition of their proliferation capacity. In addition, morphologic alterations exemplified by detachment of the cells from their solid support associated with melanin release were observed in the B-16-F10 cells. Less metastatic foci developed in the lungs of the mice treated with the purified IgG fraction from the sera of patients with vitiligo compared with those treated with purified IgG fraction from healthy subjects. CONCLUSIONS: The results of this study point to the presence of anti-melanoma autoantibodies in the sera of patients with localized and diffuse vitiligo. These antibodies have a destructive effect on melanoma cells in vitro and in vivo. PMID- 8402451 TI - Malignant cutaneous melanoma associated with neurofibromatosis in two sisters from a family with familial atypical multiple mole melanoma syndrome. Case reports and review of the literature. AB - Two cases of malignant melanoma associated with neurofibromatosis in two first degree female relatives from a family with familial atypical multiple mole melanoma (FAMMM) syndrome are presented. The types of neurofibromatosis and the FAMMM syndrome are discussed in relation to these cases and the family genealogic tree. Although the FAMMM syndrome could probably be seen as the underlying disease in the current cases, review of literature has failed to establish a clear relation. Research into pigmentary disturbance in neurofibromatosis is necessary to give a final explanation. To our knowledge, this is the first report in literature describing the familial occurrence of both diseases and it might present an addition to the tumor spectrum in the FAMMM syndrome. PMID- 8402452 TI - Diagnostic value of cerebrospinal fluid cytology in comparison with tumor marker activity in central nervous system metastases secondary to breast cancer. AB - BACKGROUND: Central nervous system (CNS) metastases occur in approximately 35% of patients with breast cancer. Parenchymal brain metastases (MET) remain undetected in a large proportion of patients, and only 50% of patients with leptomeningeal carcinomatosis (MC) are diagnosed in vivo. METHODS: Cytology and activity of the tumor markers tissue polypeptide antigen (TPA) and creatine kinase-BB isoenzyme (CK-BB) were evaluated in the cerebrospinal fluid (CSF) in 71 consecutive patients with breast cancer suspected for CNS metastases. RESULTS: Forty-three patients had no CNS metastases, 12 patients had MET, 5 patients had both MET and MC, and 11 patients had MC alone. Seven of the patients with MC had an intracerebroventricular (ICV) reservoir inserted, and an additional 70 ICV CSF samples from these patients were obtained. In CSF obtained by lumbar puncture, 11% of the samples were classified as "suspicious for malignancy," but a very limited interobserver variability was demonstrated (Kappa test value, 0.81; 95% confidence limits, 0.67-0.95%). Fifty-one percent of the ICV CSF samples were classified as "suspicious for malignancy" (Kappa test value, 0.58; 95% confidence limits, 0.34-0.82%). TPA and CK-BB were both measured in 101 CSF specimens (61 from lumbar puncture and 40 ICV samples). The differences between patients with and without CNS metastases were significantly different according to TPA (P < 0.00001) and CK-BB (P < 0.00003; Mann-Whitney test). The sensitivity and predictive value of a negative test for having any CNS metastases (in case of elevated values of either TPA or CK-BB or both) were 85% (95% confidence limits, 65-96%) and 90% (95% confidence limits, 76-97%), respectively. In addition, a significant correlation between TPA and CK-BB was demonstrated in CSF from lumbar puncture (Spearmans-Rho, 0.49; P < 0.0001) and ICV (Spearmans-Rho, 0.37; P < 0.02). CONCLUSIONS: Cytologic evaluation of CSF obtained by lumbar punctures is a reliable procedure. In CSF from ICV reservoirs, cytologic evaluation is of limited use, but CK-BB and TPA is of potential value. PMID- 8402453 TI - Significance of computed tomography-measured volume in the prognosis of cervical carcinoma. AB - BACKGROUND: Computed tomography (CT) can visualize the volume and shapes of the deep-seated tumors and is expected to predict the treatment outcome in a more quantitative manner. This study was undertaken to learn the prognostic significance of CT-provided information in the radiation therapy of carcinoma of the cervix. METHODS: One hundred thirty-one CT evaluations were systematically applied to 87 patients having carcinoma of the cervix uteri. The cervical mass volume was measured by CT at a certain phase or phases of treatment and was analyzed in relation to their local control at 3 years and the survival at 5 years. RESULTS: The initial volume, measured at the beginning of radiation therapy, was not a significant prognostic guide for the local control. However, the second volume, the volume of cervical lesion measured immediately after the completion of external irradiation, proved to be a prominent prognostic factor for both the 3-year local control and 5-year survival, regardless of the clinical stage. In 52 patients whose second volume was less than 38 cm3, 47 patients (90.4%) were locally controlled for 3 years or more after radiation therapy, whereas 17 of 23 patients (73.9%) whose second volume exceeded 38 cm3 failed in the local control. Five-year actuarial survival rate was 53.1% plus or minus 6.2% in the former group and 26.1% plus or minus 9.6% in the latter group. CONCLUSIONS: CT-aided volumetry provides for an early prediction of treatment failure in radiation therapy for cervical carcinoma. PMID- 8402454 TI - Prognostic factors in advanced stage squamous cell cancer of the cervix. AB - BACKGROUND: This study was undertaken to review prognostic factors for advanced squamous cell cancer of the cervix. METHODS: A clinicopathologic review of patients diagnosed with advanced stage squamous cell cancer of the cervix was performed at the University of Michigan Medical Center, Ann Arbor, Michigan, from 1970-1985. RESULTS: All patients had squamous cell disease and were divided according to the following stages: Stage IIIa (4), Stage IIIb (113), Stage IVa (32), and Stage IVb (26). The cumulative 5-year survival was as follows: Stage IIIa (50%), Stage IIIb (37%), Stage IVa (14%), and Stage IVb (4%). Prognostic features for Stage IIIb disease showed that the intravenous pyelogram status significantly predicted cumulative 5-year survival (P = 0.00001). When the intravenous pyelogram was normal, 47% survived. When ureteral obstruction was present without renal failure, 29% survived, and when renal failure occurred, all patients were dead of disease by 16 months. The lymph node status significantly influenced cumulative 5-year survival (P = 0.004). When lymph nodes were negative, 47% survived. When three or fewer were positive, 44% survived. When more than three were positive, 11% survived. When pelvic lymph nodes were positive and paraaortic node status was determined, 25% survived when paraaortic lymph nodes were negative, while 8% survived when these were positive (P = 0.06). Factors that did not influence 5-year survival included one or both sidewall involvement (P = 0.77), tumor grade (P = 0.23), diabetes (P = 0.92), hypertension (P = 0.85), and obesity (P = 0.47). The diagnosis of Stage IVa disease was made by the presence of fistula at initial presentation (n = 8), cystoscopy (n = 21), and sigmoidoscopy (n = 1). One patient developed a treatment-related vesicovaginal fistula when bladder involvement was diagnosed by cystoscopy. All 18 patients who presented with renal failure (Stage IIIb, 9; Stage IVa, 9) were analyzed as a group, and only 1 patient survived. The median survival in 15 patients who underwent nephrostomy was 8 months, range 1-36 months. Ten of 15 patients (66%) were dead of disease within 1 year. Three patients refused renal bypass, and these three patients died at 1, 2, and 3 months, respectively. CONCLUSIONS: Advanced stage disease represents a significant challenge, and when ureteral obstruction or renal failure is present, the prognosis is markedly decreased. PMID- 8402455 TI - Prognostic importance of histologic vascular density in cervical cancer treated with hypertensive intraarterial chemotherapy. AB - BACKGROUND: Over the past 20 years, various methods have been suggested to determine the histopathologic stage and grade of cervical cancer for predicting biologic behavior during chemotherapy. However, none of these complicated techniques have proved consistently accurate enough to predict chemotherapeutic effects on locally advanced cervical cancer. METHODS: Primary treatment with hypertensive intraarterial chemotherapy and, 4 weeks later, with radical hysterectomy were performed on 20 patients with locally advanced cancer of the uterine cervix (International Federation of Gynecology and Obstetrics [FIGO] Stages IIa-IVa). The vascular densities of biopsy specimens from pretreatment cancers stained by the Elastica-Goldner technique were measured semiquantitatively. Cytologic and histologic changes in the early stages after the intraarterial chemotherapy were simultaneously recorded. Patients were divided into two groups after analysis of the final histologic effects on operation materials in accordance with the criteria of the Japan Society for Cancer Therapy: (1) an effective group (> or = Grade 2; n = 7) or (2) a noneffective group (< Grade 2; n = 13). RESULTS: The vascular densities of the effective and noneffective groups were 6.91 plus or minus 4.51/mm2 and 1.94 plus or minus 1.21/mm2, respectively. The vascular densities of biopsy specimens were significantly higher in the effective group (P < 0.05). The early cytologic and histologic changes 1 week after chemotherapy were significantly greater in the effective group (P < 0.05). CONCLUSIONS: These results indicate that the efficacy of intraarterial chemotherapy for cervical cancers depends on their individual vascular densities, which can be detected beforehand through the study of biopsy specimens. Furthermore, cytologic and histologic changes in the early stages would seem to reflect the overall histologic effect of the treatment as a whole. PMID- 8402456 TI - Differential values of Ki-67 index and mitotic index of proliferating cell population. An assessment of cell cycle and prognosis in radiation therapy for cervical cancer. AB - BACKGROUND: Little is known about correlations between the growth fraction determined immunohistochemically with Ki-67 antibody and radiation response or prognosis after radiation therapy. METHODS: The prognostic value of the growth fraction determined by Ki-67 index and the mitotic index of proliferating cell population (pMI) were assessed in 45 cervical cancers treated with radiation therapy. The specimens from the cervix before radiation therapy were immunohistochemically stained with anti-Ki-67 antibody. RESULTS: The mean Ki-67 index and pMI for all patients were 36.0% and 2.74%, respectively. The patients with a Ki-67 index of 33% or greater showed significantly better histologic response to radiation at 30 Gy than those with less than 33%. The mean Ki-67 index for patients with good prognosis was significantly higher than for patients with tumor recurrence or metastasis later. Further, the mean values of pMI for patients with good prognosis were significantly lower than for patients with recurrence or metastasis. The 3-year survival rate for higher Ki-67 index (> or = 33%) was significantly better than lower Ki-67 index (less than 33%) (90.9% versus 34.8%; P < 0.001). However, the 3-year survival rate for higher pMI (> or = 3.5%) was significantly poorer than lower pMI (less than 3.5%) (8.3% versus 81.8%; P < 0.001). CONCLUSIONS: These results suggested that tumors with a high growth fraction showed a good prognosis with radiation therapy. In addition, the inverse prognostic correlation between the Ki-67 index and pMI suggested that both indices have independent values on radiation response and prognosis after radiation therapy. PMID- 8402457 TI - High-dose rate and low-dose rate intracavitary therapy for carcinoma of the uterine cervix. Final results of Osaka University Hospital. AB - BACKGROUND: High-dose rate (HDR) intracavitary radiation therapy for carcinoma of the uterine cervix has gradually found wider acceptance. In 1983, the authors first presented the results of prospective randomized comparative study of HDR versus low-dose rate (LDR) therapy. In the current study, the final results of this study with a longer follow-up are presented. METHODS: From January 1975 through August 1983, 430 previously untreated patients with carcinoma of the uterine cervix in Stages I-III were treated with either HDR 60Co therapy or LDR 137Cs therapy at our department. HDR was administered to a total of 259 patients: 32 patients in Stage I, 80 in Stage II, and 147 in Stage III. LDR was administered to a total of 171 patients: 28 patients in Stage I, 61 in Stage II, and 82 in Stage III. RESULTS: The 5-year cause-specific survival rates of Stage I III patients treated with HDR were 85%, 73%, and 53%, respectively. The corresponding figures for LDR were 93%, 78%, and 47%, respectively. There was no significant difference between these survival rates. Moderate-to-severe complications developed in 10% of the patients treated with HDR and 4% of those with LDR. This difference in the incidence of complications was statistically significant (P = 0.023). CONCLUSIONS: Treatment results in terms of cause specific survival were equivalent for HDR and LDR treatment. However, the incidence of complications was higher for the HDR group, although within acceptable levels, than for the LDR group. PMID- 8402458 TI - The prognostic value of tetranectin immunoreactivity and plasma tetranectin in patients with ovarian cancer. AB - BACKGROUND: Tetranectin (TN), a tetrameric, plasminogen-binding protein, was reduced in the plasma of patients with cancer and appears extracellularly in "stimulated" connective tissues, such as the proliferative, connective tissue response to carcinomas known as desmoplasia. METHODS: Tissue samples from 37 patients with ovarian cancer were examined immunohistochemically for stromal and cellular TN. Plasma samples obtained before the primary surgery were quantitated for TN. The univariate log-rank test and the multivariate Cox proportional hazards regression model were used to analyze the prognostic function of the variables. RESULTS: A significantly higher survival rate was found for patients with a low-stromal TN score and a high-plasma TN concentration, whereas the cellular TN score did not have any significance. A significant negative correlation was found between plasma TN and stromal TN (RS = -0.36; P = 0.03). Independent significant correlations were found between stromal immunoreactivity for TN and tumor grade (R = 0.67; P = 0.03) and between plasma TN and tumor stage (R = -0.29; P = 0.01). CONCLUSIONS: This study gives great expectations to TN as a useful parameter for prognostic evaluation of patients with ovarian cancer. According to the correlations, stromal TN may partly originate from plasma and enhance proteolytic degradation in the interstitial tissue, a process necessary for the spread and growth of cancer. Because plasma TN measurements are only valid when taken preoperatively, it is of great value that stromal TN immunoreactivity of stored tumor tissue can be used as a prognostic factor for ovarian cancer. PMID- 8402459 TI - Loss of constitutional heterozygosity on chromosome 11p in human ovarian cancer. Positive correlation with grade of differentiation. AB - BACKGROUND: There is increasing evidence suggesting that genes located on the short arm of chromosome 11 play an important role in the development of human ovarian cancer. Recent cytogenetic and molecular studies have demonstrated the loss of genetic material in this region. Loss of normal growth regulatory genes may allow for the expression of tumorigenicity or lead to tumor progression. METHODS: The authors used DNA recombinant techniques to examine the frequency of allelic losses at four loci spanning the chromosomal region 11p15.1-11p15.5 in 40 patients with malignant ovarian tumors. DNA extracts from normal leukocytes and 48 tumor samples were analyzed by Southern blotting using the polymorphic probes pEJ6.6 (HRAS1), phins310 (INS), p20.36 (PTH), and pEM36 (CALCA). RESULTS: Reduction to homozygosity in the tumor DNA was found in 47.5% of the informative cases (19 of 40). Comparing the results with clinical parameters, none of the well-differentiated tumors (6 of 40, Grade 1) and only one of the early stage tumors (6 of 40, International Federation of Gynecology and Obstetrics [FIGO] Stage I or II) showed alterations in this chromosome region. Statistical analysis revealed a strong correlation of rate of loss of constitutional heterozygosity (LOH) and grade of differentiation, in the sense of higher 11p allele losses occurring in poorly differentiated tumors. CONCLUSIONS: The authors concluded that the relatively high incidence of 11p allele losses marks an important step in ovarian cancer development. Furthermore, statistical analysis showed that loss of 11p alleles was strongly correlated with poorly differentiated ovarian cancer, indicating the location of genes involved in cellular functions associated with the development of more anaplastic tumors. PMID- 8402460 TI - Serum and ascitic fluid levels of interleukin-1, interleukin-6, and tumor necrosis factor-alpha in patients with ovarian epithelial cancer. AB - BACKGROUND: Recent studies have shown that multiple cytokines are secreted by ovarian epithelial cancer cells. Previous studies have shown that the cancer cell lines secrete macrophage colony-stimulating factor (M-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-1 (IL-1), interleukin 6 (IL-6), and transforming growth factor-alpha (TGF-alpha). Concomitantly, the serum levels of one of the growth factors (M-CSF) was found to be significantly elevated in patients with primary ovarian cancer and in second-look patients. The authors evaluated the serum levels of IL-1 alpha, IL-1 beta, IL-6, and tumor necrosis factor-alpha (TNF-alpha) in patients with primary ovarian epithelial cancer. These levels were then compared with cytokine concentration found in normal peritoneal fluid. METHODS: Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of cytokines in normal peritoneal fluid, ascites, and serum. RESULTS: In serum, TNF-alpha and IL-6 were significantly increased in primary ovarian cancer patients when compared with control subjects (P < 0.0001 for both cytokines). TNF-alpha and IL-6 were also significantly higher than the levels found in second-look patients (P < 0.007 for TNF-alpha, and P = 0.0002 for IL-6). The levels of IL-1 alpha and beta were not elevated in ovarian cancer. TNF alpha in the ascites was higher when compared with normal peritoneal fluid and was statistically significantly different when a cut-off point between 71-110 pg was selected (P < 0.005). The levels of IL-6 in ascites from patients with primary ovarian cancer also showed a marked increase (P < 0.0001) when compared with peritoneal fluid from control subjects. CONCLUSIONS: Levels of IL-1, IL-6, and TNF-alpha were determined in normal peritoneal fluid, ovarian malignant ascites, normal serum, and serum from patients with ovarian cancer. This study showed that the patients with ovarian cancer have elevated levels of IL-6 and TNF alpha in serum and ascitic fluid. A larger study would help in evaluating the potential use of cytokines as tumor markers in ovarian cancer. PMID- 8402461 TI - Ovarian carcinomas with choriocarcinomatous differentiation. AB - BACKGROUND: Ovarian carcinomas may produce human chorionic gonadotropin (HCG) or HCG-like substances and may even contain syncytiotrophoblast cells, but a true choriocarcinomatous component has not been described in these tumors. METHODS: Two cases of poorly differentiated ovarian carcinoma with choriocarcinomatous components are reported. Pathologic findings were correlated with immunohistochemical stains, hormonal effects, and clinical behavior. RESULTS: Each tumor contained a circumscribed, extensively necrotic and hemorrhagic brown nodule. Microscopically, the nodules exhibited a mixture of cytotrophoblast and syncytiotrophoblast. The syncytiotrophoblast capped cytotrophoblast and was strongly positive for beta-HCG. In one of the cases, a transformation zone composed of poorly differentiated carcinoma with occasional beta-HCG-positive cells was observed between a mucinous cystadenoma and the choriocarcinomatous elements. The two cases exhibited activation of the ovarian stroma in the form of condensation and luteinization. Extra-abdominal metastases developed early in both patients and, despite multiagent chemotherapy, they died shortly postoperatively. CONCLUSIONS: Choriocarcinoma may rarely develop as a result of dedifferentiation of common epithelial ovarian tumors. Recognition of choriocarcinomatous components in ovarian carcinomas is important because of its association with aggressive behavior. PMID- 8402462 TI - Transitional cell carcinoma pattern in primary carcinoma of the fallopian tube. AB - BACKGROUND: A broad papillary proliferation resembling that in transitional cell carcinoma (TCC) of the urinary bladder was seen in 12 of 21 primary carcinomas of the Fallopian tube (PCFT). METHODS: According to their predominant histologic pattern (more than 50%), PCFT were classified into 9 TCC-predominant and 12 non TCC-predominant tumors. The two groups were compared by clinicopathologic, histochemical, and immunohistochemical means. RESULTS: TCC-predominant tumors were grossly solid and microscopically demonstrated more frequent tumor necrosis and spindled tumor cells than non-TCC-predominant tumors. Mucin histochemistry revealed a correlation between TCC-predominant tumor and sulfomucin-predominant secretion and between non-TCC-predominant tumor and sialomucin-predominant secretion. Immunohistochemical studies for cytokeratins, vimentin, epithelial membrane antigen (EMA), Leu-M1, carcinoembryonic antigen (CEA), and CA 125 were not useful for discrimination between the two groups. Both groups showed similar features in patient age, clinical stage, cytology of ascites or peritoneal washing, and serum CA 125 level. Despite the similarity in treatment (surgery and postoperative chemotherapy) between the two groups, TCC-predominant tumors tended to relapse later (mean, 31.2 months after diagnosis) than non-TCC-predominant tumors (mean, 14.4 months after diagnosis), resulting in a significant difference in the 2-year disease-free survival rate. CONCLUSIONS: TCC pattern and non-TCC pattern are considered to be worthy of distinction in PCFT. PMID- 8402463 TI - Bladder cancer risk assessment with quantitative fluorescence image analysis of tumor markers in exfoliated bladder cells. AB - BACKGROUND: The detection of potentially highly curable low-grade bladder cancers by noninvasive techniques remains an unsolved problem. Conventional cytology detects such tumors with 50% sensitivity, and addition of DNA measurements to cytology only improves sensitivity incrementally. Tumor-associated antigens potentially offer an additional diagnostic marker. METHODS: In this study, the M344 antibody against a tumor-associated antigen expressed mainly by low-grade tumor cells was tested for its sensitivity and specificity, alone and in combination with DNA ploidy and cytology. Voided urine samples from 69 asymptomatic control subjects, urines and bladder washings from 59 patients with cancer, and 195 symptomatic control patients were collected. Cells were double labeled with M344 monoclonal antibody and Hoechst. Each case was blinded, and the number of positive cells was scored by two independent observers. RESULTS: High grade and low-grade transitional cell carcinomas (TCC) were detected with equal efficiency (78%, P < 0.001 versus symptomatic control patients). Urine samples proved higher specificity in detecting cancers. Patients being monitored for recurrence, but without current detectable cancer, were intermediates between control subjects and patients with cancer, suggesting that this marker also responds to dysplasia or field disease. Patients with outlet obstruction did not significantly differ from patients with previous TCC (P = 0.95). When combined with DNA ploidy measurements and cytology, the sensitivity for low-grade and high grade tumors was 88% and 95%, respectively. CONCLUSIONS: The M344 antibody potentially could improve the specificity and sensitivity of detection of low grade bladder tumors in symptomatic and asymptomatic patients as well as monitoring for recurrence, therapeutic response, and assessment of individual risk. PMID- 8402464 TI - Quantitative histopathology in the prognostic evaluation of patients with transitional cell carcinoma of the urinary bladder. AB - BACKGROUND: Morphologic grading of malignancy is considered to be of prognostic value in patients with transitional cell carcinomas of the urinary bladder (TCC). This qualitative approach is, however, associated with low reproducibility. Grading of malignancy can be carried out on a reproducible, quantitative scale. METHODS: A retrospective, prognostic study of 110 patients treated for TCC in clinical Stages Ta-T4 (median follow-up time, 6 years) was performed, evaluating various grading techniques. Unbiased estimates of the volume-weighted mean nuclear volume (nuclear vV), nuclear volume fraction, estimates of nuclear mean profile area (aH(nuc)), nuclear profile density index (NI), and mitotic profile density index (MI) were obtained by stereologic and morphometric techniques. RESULTS: The T-stage and morphologic grade of malignancy were closely cross correlated (+0.63 < Kendall tau < +0.71, 2P < 6.7 x 10(-16)). The estimation of nuclear vV was highly efficient, with more than 85% of the associated variation attributable to differences between tumors. A positive significant correlation between estimates of nuclear vV and aH(nuc) was detected (r = +0.79), whereas an inverse correlation was documented between nuclear vV and NI (r = -0.63). Estimates of nuclear volume fraction showed no correlation with nuclear vV. Comparisons between categorical and quantitative data revealed the following: a decrease in averaged estimates of NI for tumors in advanced T-stage and malignancy grade (2P < 0.0008); and nuclear vV and aH(nuc) increased on average, in tumors of high T-stage and malignancy grade. Estimates of MI were also positively correlated with the T-stage and the malignancy grade (+0.42 < Kendall tau < +0.49). Single-factor analyses showed prognostic effect of T-stage, grade of malignancy, and, apart from nuclear volume fraction, all quantitative histopathologic variables with regard to overall survival (2P < 0.03). None of the morphometric and stereologic parameters were of prognostic values with regard to recurrence-free survival (2P > 0.26). Multiple hazards regression analysis (Cox models) revealed that clinical stage of disease was the sole independent prognostic variable. Only estimates of nuclear vV added significant independent prognostic prediction with regard to recurrence-free survival in the 48 patients with Ta tumors (2P = 0.03). CONCLUSIONS: The results suggested that estimates of nuclear vV are prognostically superior to morphologic grading of malignancy in noninvasive TCC, whereas both morphologically and quantitatively based malignancy grading are without prognostic value in invasive TCC. PMID- 8402465 TI - Heart involvement in lymphomas. The value of magnetic resonance imaging and two dimensional echocardiography at disease presentation. AB - BACKGROUND AND METHODS: Thirty-six patients with mediastinal lymphoma were studied with chest magnetic resonance imaging (MRI) and two-dimensional echocardiography at presentation to define the extent of the disease in the paracardiac area. RESULTS: Involvement of cardiac structures was present in 23 of 36 patients (64%). Pericardial contiguity was detected in 23 of 23 patients (100%) by MRI and in 18 of 23 patients (78%) by echocardiography. Pericardial effusion, present in 17 patients (74%), and pericardial infiltration, present in 7 patients (30%), were detected by both techniques in 71% and 86%, respectively. Myocardial infiltration was identified in two of two patients (100%) by MRI and in one of two patients (50%) by two-dimensional echocardiography. Extrapericardial disease was identified in 100% of patients by MRI but only in 30% of patients by echocardiography. CONCLUSIONS: Extracardiac and intracardiac involvement is a frequent event in mediastinal lymphomas and should be carefully evaluated with different imaging modalities, mainly MRI, for correct diagnosis and proper management. PMID- 8402466 TI - Reactivity of B72.3 with adenocarcinomas. An immunohistochemical study of 476 cases. AB - BACKGROUND: The monoclonal antibody B72.3 has been shown to react with several common types of adenocarcinoma. However, the distribution of B72.3 reactivity in adenocarcinomas has not been well-enough defined to determine whether B72.3 immunostaining would be helpful in the diagnosis of adenocarcinoma of unknown origin. METHODS: Formaldehyde fixed, paraffin-embedded tissue from 476 adenocarcinomas from a variety of primary sites were studied using B72.3 and an avidin-biotin complex immunohistochemical technique. RESULTS: B72.3 immunoreactivity was seen in 75-100% of the adenocarcinomas from the ovary, endocervix, endometrium, distal esophagus/stomach, colon, lung, breast, prostate, pancreas, and bile ducts. Positive reactions were unusual in thyroid carcinomas (7%) and renal cell carcinomas (11%). B72.3 reactivity was not seen in the 20 hepatocellular carcinomas or 4 adrenal cortical carcinomas studied. CONCLUSIONS: B72.3 reacts with a wide variety of adenocarcinomas. Immunostaining with B72.3 may be helpful in excluding hepatocellular carcinoma in certain clinical settings. PMID- 8402467 TI - Congenital mesoblastic nephroma metastatic to the brain. AB - BACKGROUND: Congenital mesoblastic nephroma (CMN) is generally considered to be a benign neoplasm requiring only total excision. Rare local recurrences have usually been related to incomplete removal, and distant metastases to the lung have been reported three times. METHODS: The authors reported the first case of CMN metastatic to the brain, illustrating a comparison of the histopathology of the primary and secondary lesions. RESULTS: The gross appearance of the renal tumor was that of a benign CMN. On histopathologic examination, there was a single nodule of more cellular tissue with focal necrosis. This area was diagnosed as within the designation "cellular" mesoblastic nephroma. The appearance of the brain metastasis corresponded to that of the cellular nodule. CONCLUSIONS: This report expands the spectrum of metastatic potential of CMN. Though a rare event, this case underscores the need for close follow-up of infants with CMN. PMID- 8402468 TI - Prognostic features of Ewing sarcoma on plain radiograph and computed tomography scan after initial treatment. A Pediatric Oncology Group study (8346) AB - BACKGROUND: The authors studied the short-term changes in the plain radiographic and computed tomography (CT) appearance of Ewing sarcoma for indicators of decreased survival or future disease progression. METHODS: The authors evaluated CT scans and plain radiographs of the primary tumor site from 105 patients with Ewing sarcoma at diagnosis (prebiopsy), after induction chemotherapy (13 weeks), and after radiation therapy (20 weeks). RESULTS: Data suggest an association between postinduction CT findings of medullary involvement, cortical destruction, lysis, permeation, and unhealed pathologic fracture and decreased survival. On the postradiation scans, only medullary involvement was associated with worsened survival. No plain radiographic features were significant at any time. Absolute greatest tumor dimension was not significantly related to survival or tumor progression. The Cox model suggested that fractional change in greatest tumor dimension on CT at the time points studied relative to the prebiopsy CT was correlated to survival. Log-rank testing did not corroborate this finding. All significant associations appeared to result from adverse outcomes in small subgroups. CONCLUSIONS: Our data suggest that CT obtained immediately after induction chemotherapy and radiation may have some limited use in predicting the long-term prognosis of patients with Ewing sarcoma. PMID- 8402469 TI - Undifferentiated (embryonal) sarcoma of the liver in childhood. Successful combined-modality therapy in four patients. AB - BACKGROUND: Undifferentiated (embryonal) sarcoma of the liver is a rare, highly malignant, mesenchymal tumor presenting predominantly in late childhood. Four girls, ages 6-13 years, who were treated with combined-modality regimens are reported. METHODS: In the first and second patients, hemihepatectomy resulted in complete removal of the tumor, and multiple-agent chemotherapy was administered postoperatively. In the third patient, only partial resection could be accomplished initially. By synchronous radiation therapy and chemotherapy, the tumor decreased to an extent that it could be resected completely and was totally devitalized on histologic examination. Postoperative chemotherapy was discontinued after 8 weeks. In the fourth patient, the tumor was not resectable at diagnostic biopsy. Polychemotherapy led to a significant reduction of the tumor size, and resection with clear margins could be performed subsequently. Because histologic necrosis amounted to about 95%, postoperative chemotherapy was also discontinued after 6 weeks. RESULTS: All four patients remain well without evidence of tumor recurrence after 79, 41, 36, and 22 months from diagnosis. CONCLUSIONS: The authors suggest that a multimodal therapeutic regimen should be used in patients with undifferentiated hepatic sarcoma. PMID- 8402470 TI - Risk-directed therapy for childhood acute lymphoblastic leukemia. Results of the Associazione Italiana Ematologia Oncologia Pediatrica '82 studies. AB - BACKGROUND: In 1982, the Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) started its third-generation study, aiming to improve previous results obtained by AIEOP '79 study and to deliver a standardized treatment to most Italian children with acute lymphoblastic leukemia (ALL). METHODS: We treated 902 children (older than 1 year and younger than 15 years of age) with newly diagnosed ALL in multicenter studies of risk-directed therapy (111 low risk [LR] from Study 8201; 570 average risk [AR] from Study 8202; and 117 and 104 high risk [HR] from Studies 8303 and 8503, respectively). Induction therapy was composed of vincristine, prednisone, and asparaginase for LR or AR patients and these agents plus daunorubicin, (Study 8503) or vincristine, prednisone, cytarabine, and intermediate-dose methotrexate (Study 8303) for HR patients. Central nervous system (CNS) preventive therapy consisted of intrathecal methotrexate only (LR), intrathecal methotrexate plus 18 Gy cranial irradiation (AR and HR Study 8503), or high-dose (HD) cytarabine (HR Study 8303). Reinduction therapy was vincristine/prednisone/daunorubicin for AR patients with cyclophosphamide added for HR patients in Study 8303 and HD asparaginase in Study 8503. LR patients did not receive intensification therapy. Continuation therapy comprised 6 mercaptopurine plus methotrexate and monthly pulses with vincristine plus prednisone for all patients, except for HR patients in Study 8303 who also received teniposide plus cytarabine. Weekly HD asparaginase was also given in Study 8503. Duration of treatment was 24 months for Studies 8201 and 8202, 15 months for Study 8303, and 22 months for Study 8503. The overall complete remission (CR) rate was 94.7% (97.3% for LR, 94.9% for AR, and 93.2% for HR). RESULTS: Overall 7-year event-free survival (EFS) was 53.6% (standard error [SE], 1.8). EFS was 60.8% in LR (SE, 4.7), 60.6% in AR at 7 years (SE, 4.7), and 18.5% in Study 8303 (HR) at 5 years (SE, 3.8). Because of the poor result in HR patients, a successor study (8503) was developed that yielded a 5-year EFS of 46.1% (SE, 5.1). Site-specific relapse rates were 18.5% (LR), 13.4% (AR), 35.1% (HR on 8303), and 18.3% (HR in Study 8503) for bone marrow and 9.2%, 7.9%, 17.5%, and 19.3%, respectively, for the CNS (isolated). Isolated testicular relapse was observed in 3.9% of male patients. CONCLUSIONS: This risk-directed therapy cured at least 50% of patients with ALL with relatively nonintensive therapy. The 80% overall survival rate for LR and AR patients at 7 years suggested that the total cure rate may be higher than 50% because of the significant salvage rate for patients who received antimetabolite-based therapy initially. PMID- 8402471 TI - Screening for human papillomavirus infections in esophageal squamous cell carcinomas by in situ hybridization. AB - BACKGROUND: Infections with specific types of human papillomavirus (HPV) have been closely linked with human squamous cell carcinomas, those of the anogenital tract in particular. Increasing number of reports also suggest that HPV infection could be a risk factor for esophageal cancer. However, most of the previous studies on HPV involvement in esophageal carcinomas have included only small numbers of biopsy specimens, thus necessitating additional studies based on extensive series of esophageal samples. METHODS: A series of 776 biopsy specimens derived from 363 patients who had undergone esophagectomy for squamous cell carcinoma in the high-incidence area of China were analyzed for the presence of HPV DNA by screening and specific typing in situ hybridization with biotinylated HPV DNA probes. RESULTS: Under low-stringency conditions, 85 (23.4%) tumors were demonstrated to contain HPV DNA: Positive signals were found on the nuclei of cancer cells in 71 (19.6%), in the surrounding epithelial cells with hyperplastic or dysplastic changes in 13 (3.6%), in the cancer cells and the surrounding epithelial cells in 10 (2.8%), and in the resected margins in 1 (0.3%). Thirty four (40%) of the 85 HPV-positive tumors were shown to contain at least one type of HPV 6, 11, 16, 18, or 30 DNA sequences. HPV 16 was the type found most frequently, occurring in 18.8% of the 85 HPV-positive specimens. In addition to the primary tumors, HPV DNA sequences were found in 12.3% (7 of 57) of the lymph node metastases. CONCLUSION: The results confirm the previously reported HPV involvement in esophageal squamous cell lesions and implicate HPV as a potential etiologic agent in the multifactorial pathogenesis of esophageal carcinoma. PMID- 8402472 TI - Risk factors for esophageal cancer in women in northern Italy. AB - BACKGROUND: The incidence of esophageal cancer in women in Italy is low, and its risk factors have not been studied extensively. METHODS: The relationship between risk of esophageal cancer and frequency of consumption of tobacco, alcohol, and a few selected dietary items was studied in 57 Italian women with histologically confirmed incident cancers of the esophagus and 344 hospital control patients, using data from a case-control study conducted in Milan, Italy, 1984-1991. RESULTS: The major risk factor for cancer of the esophagus in Italian women was cigarette smoking (relative risk [RR], 1.5 for < 15 cigarettes/day, 95% confidence interval [CI], 0.6-3.8; and RR, 4.8 for > or = 15 cigarettes/day, 95% CI, 2.2-10.3; compared with those who never smoked), followed by elevated alcohol consumption (RR, 2.3; 95% CI, 1.0-5.4 for three or more drinks/day, relative to teetotalers), which together explained more than 50% of cases. Among dietary items, high intake of fresh fruit showed a statistically significant protective effect (RR, 0.4; 95% CI, 0.2-0.9 for the highest versus the lowest tertile of intake). There was an inverse relationship with an estimate of beta-carotene intake (RR, 0.5; 95% CI, 0.2-1.0 for the highest versus the lowest level of intake). No association was evident with preformed vitamin A (retinol). CONCLUSION: On a population scale, tobacco is the major risk factor for esophageal cancer in Italian women. Although the incidence of esophageal cancer is much lower in women than in men, major risk and protective factors are similar for both sexes. PMID- 8402473 TI - High-dose chemotherapy with autologous bone marrow transplantation in the treatment of advanced gastric cancer. AB - BACKGROUND: Clinical usefulness of high-dose chemotherapy with autologous bone marrow transplantation (ABMT) in the treatment of gastric cancer was investigated. METHODS: Ten patients with late-stage gastric cancer (four recurrent and six primary cases) with measurable lesions were treated with high dose combination therapy consisting of etoposide (VP-16) (1200 mg/m2), adriamycin (80 mg/m2), and cisplatin (120 mg/m2) or carboplatin (900 mg/m2) (high-dose etoposide, adriamycin, and cisplatin [EAP] therapy). Afterward, 1 x 10(7)/kg of mononuclear cells, which were prepared from autologous bone marrow before treatment and cryopreserved, were transfused intravenously. RESULTS: All patients recovered from the aplastic period without severe complications. Of nine assessable cases, partial response was observed in eight (89% response rate). CONCLUSION: High-dose EAP therapy with ABMT is recommended as adjuvant therapy accompanied with radical operation for late-stage gastric cancer. PMID- 8402474 TI - Do occupational factors influence the risk of colon and rectal cancer in different ways? AB - BACKGROUND: Occupational exposures and physical activity have been considered as risk factors for the development of colorectal cancer. METHODS: A case-control study on working conditions and the risk of colon and rectal cancer was performed in southeastern Sweden during 1984-86. Involved were 177 patients, 98 with colon cancer and 79 with rectal cancer, and two groups of control subjects, 371 hospital control subjects and 430 population control subjects. RESULTS: A significantly decreased risk of left-sided colon cancer was observed in persons involved in more than 20 years of physically active work and a significantly decreased risk of rectal cancer in persons involved in more than 20 years of sedentary work. A tendency toward increased risk was seen for colon cancer in male railroad workers and in male gas station workers. A reduced risk of rectal cancer was found for drivers, textile workers, and administration workers, whereas an increased risk of rectal cancer appeared among paper workers and assistant nurses. A low risk of both colon and rectal cancer was found among construction workers and forestry workers. Exposure to asbestos carried a slightly increased risk of colon cancer, whereas exposure to solvents slightly decreased the risk of rectal cancer. CONCLUSION: This study confirms earlier findings that physical activity decreases the risk for left-sided colon cancer, but also suggests that occupational factors influence the risk of colon and rectal cancer in different ways. PMID- 8402475 TI - Small encapsulated hepatocellular carcinoma of the liver. Provisional analysis of pathogenetic mechanisms. AB - BACKGROUND: Small hepatocellular carcinomas frequently were found incidentally during routine pathologic examinations of adult livers removed at liver transplant. METHODS: Sixty-nine carcinomas of all sizes were found in 25 patients; 39 of the tumors were smaller than 1 cm in diameter, and 18 of the carcinomas in five patients were not clinically suspected. These small incidental carcinomas lend themselves to analysis of the morphologic basis of human hepatocellular carcinogenesis. RESULTS: All of these tumors arose in cirrhotic livers. Most of the small carcinomas were multilobulated and subdivided by pre existing fibrous septa. The surrounding capsule usually was not a true capsule. They were all well differentiated, most formed bile, Mallory bodies, or showed alpha-1-antitrypsin (A1AT) positivity. Transition from cirrhotic nodular parenchyma to areas of hyperplasia or atypical hyperplasia to well-differentiated carcinoma were common. Large cell dysplasia also was common. CONCLUSIONS: These morphologic transitions closely parallel changes seen in experimental chemical carcinogenesis. They also strongly suggest a multicentric origin of the tumors. In addition, in every instance, the lesions were multiple in the liver and involved both lobes. This latter finding has possible implications for recurrence after local surgical excision of small hepatocellular carcinomas. PMID- 8402476 TI - A phase I-II trial of mitoxantrone by hepatic arterial infusion in patients with hepatocellular carcinoma or colorectal carcinoma metastatic to the liver. AB - BACKGROUND: Mitoxantrone is an anthraquinone derivative that has demonstrated encouraging preclinical and clinical activity against a variety of human carcinoma cell lines and malignancies. Three Phase II studies of systemically administered mitoxantrone in patients with colorectal carcinoma failed to demonstrate any therapeutic activity, as did four Phase II studies of intravenous mitoxantrone in hepatocellular carcinoma. Two additional trials demonstrated limited activity when administered intravenously to patients with hepatocellular carcinoma. However, because this drug exhibits a steep dose-response curve, a Phase I-II trial of mitoxantrone by hepatic arterial infusion was initiated. METHODS: Patients with hepatocellular carcinoma and metastatic colorectal carcinoma with liver only or liver-predominant disease were eligible for therapy. All patients underwent the placement of a percutaneous hepatic arterial catheter before each course of therapy, and the first cohort of patients was treated at 10 mg/m2/course on day 1 on a 28-day cycle. Dosages were escalated in increments of 2 mg/m2/course based on side effects and tolerance. RESULTS: Twenty-eight patients with bidimensionally measurable unresectable, liver-predominant disease were entered into this trial. The therapy was well tolerated, with only 5 courses of 55 being complicated by neutropenia and none associated with fever. Only one patient required a dosage reduction on the basis of toxicity (neutropenia). No complete or partial responses were observed. CONCLUSION: These data are consistent with a lack of therapeutic activity of mitoxantrone when administered by hepatic arterial infusion for the treatment of hepatocellular carcinoma or metastatic colorectal cancer. PMID- 8402477 TI - Predominant etiologic association of hepatitis C virus with hepatocellular carcinoma compared with hepatitis B virus in elderly patients in a hepatitis B endemic area. AB - BACKGROUND: It has been thought that the mean age of patients with hepatitis B surface antigen (HBsAg) positivity and hepatocellular carcinoma (HCC) is younger than that of patients who test positive for anti-hepatitis C virus. This study was to assess the relative etiologic association of hepatitis B virus (HBV) and hepatitis C virus (HCV) with HCC, according to age-specific groups. METHODS: A total of 336 Korean patients with HCC were enrolled in the study; the mean age was 53.7 +/- 9.1 years, and the male-to-female ratio was 5.9. HBV serologic markers and anti-HCV were tested using commercially available radioimmunoassay and enzyme immunoassay kits, respectively. RESULTS: Two hundred thirty-three (69.3%) patients tested positive for HBsAg; anti-HCV was positive in 42.7% of 103 patients with HBsAg-negative disease and in 17.0% of all patients with HCC. Coinfection rate of HBV and HCV in the total number of patients with HCC was 3.9%. The mean age was 51.6 years for patients with HBsAg-positive disease and 60.4 years for patients with anti-HCV-positive disease. The ratio of HBsAg positivity compared with anti-HCV positivity in patients with HCC was 29.7 for patients younger than 50 years; 0.9 for patients older than 61 years; and 5.2 for patients in their 50s. CONCLUSIONS: The authors conclude that although HBV infection plays a more important etiologic role in the development of HCC in Korea than does HCV, HCV is no less frequently involved than HBV in the etiology of HCC for patients older than 61 years. Thus, in surveillance programs for the early detection of HCC, closer attention should be directed to the patients with anti-HCV-positivity and chronic liver disease, especially patients older than 61 years, even in a hepatitis-B-endemic area. PMID- 8402478 TI - Lymphoma resembling Hodgkin disease after posttransplant lymphoproliferative disorder in a liver transplant recipient. AB - BACKGROUND: Posttransplant Epstein-Barr virus (EBV)-associated lymphoproliferative disorders (PTLD) are B-cell derived tumors of polymorphic to monomorphic histologic type. Hodgkin disease (HD) is not considered part of this spectrum and rarely occurs in the post-transplant population. A liver allograft recipient is reported who had a PTLD develop that resolved after a reduction of immunosuppression. Several years later, the patient had a separate lymphoid tumor develop that resembled HD. The relationship between the two proliferations is analyzed. METHODS: Specimens were analyzed for T- and B-cell rearrangements and EBV status by Southern blot hybridization. Immunoperoxidase studies were performed to determine the phenotypic status of the lesions. In situ hybridization for EBV early RNA (EBER) was done to determine the distribution of virus-positive cells in tissue sections. RESULTS: Unique clonal rearrangements of immunoglobulin genes and EBV were seen in each instance. No evidence of T-cell receptor rearrangement was observed. EBER positivity was widespread in the PTLD and largely, but not exclusively, limited to the anaplastic cell in the HD-like tumor. The phenotype of these anaplastic cells was similar to that described for Reed-Sternberg (RS) cells. CONCLUSION: The two tumors are unique B-cell growths associated with separate clones of lymphocytes carrying episomal EBV. Patients with PTLD may be at risk for the continued development of EBV-associated disorders. Lymphomas resembling HD represent one possible manifestation of this risk. PMID- 8402479 TI - Intradural parenchymal involvement in the spinal subarachnoid space associated with primary lung cancer. AB - BACKGROUND: Intradural parenchymal involvement (IPI) in the spinal subarachnoid space associated with primary lung cancer is rare. A retrospective study was undertaken to investigate the clinical and pathologic features of IPI. METHOD: A total of 1215 cases of primary lung cancer were studied at autopsy; the results were reviewed retrospectively. RESULTS: Twenty (1.65%) of the cases revealed IPI in the spinal subarachnoid space. The histologic diagnoses were small cell carcinoma in ten cases, adenocarcinoma in eight cases, and squamous cell carcinoma in two cases. In 14 (70%) cases, the IPI was located between the lumbar and cauda equina of the spinal cord. However, no metastases were observed in the cervical spinal cord. Brain metastasis, vertebral metastasis, and meningeal carcinomatosis were seen in 70%, 60%, and 40% of the 20 cases, respectively, suggesting that these metastases may be related to the metastatic pathway to the spinal cord. Most patients had neurologic symptoms or signs referable to IPI; IPI could be diagnosed before death in only one patient by magnetic resonance imaging. The median interval between diagnosis of lung cancer and development of IPI and median survival after the onset of neurologic symptoms referable to IPI were 415 days and 110 days, respectively. CONCLUSION: The authors retrospectively received 1215 autopsies of patients with primary lung cancer and found 20 (1.65%) with IPI. PMID- 8402480 TI - Combination therapy with interferon-dexamethasone for newly diagnosed patients with multiple myeloma. AB - BACKGROUND: Alpha-interferon and dexamethasone are each effective in patients with multiple myeloma and have a combined inhibitory effect on the in vitro growth of myeloma colonies. The effect of combined therapy in newly diagnosed patients is unknown. METHODS: Fifty-one consecutive patients with previously untreated multiple myeloma of low tumor mass received primary therapy with 3 million units (mu)/m2/day of interferon administered subcutaneously for 20 days and 20 mg/m2 of dexamethasone given orally each morning for 4 days beginning on days 1, 9, and 17. Courses were repeated after a rest period of 14 days. Results were compared with those of similar patients who received primary treatment with dexamethasone alone in the same dose regimen. RESULTS: The response rate was similar: 57% for patients treated with interferon-dexamethasone and 48% for those treated with dexamethasone alone. Remission and survival times of both groups were identical. Twenty-nine percent of patients resistant to interferon dexamethasone and 19% of patients resistant to dexamethasone responded subsequently to either standard melphalan-prednisone or to a cyclophosphamide vincristine-doxorubicin-dexamethasone combination. These regimens were also effective in one third of patients with disease relapse despite interferon. CONCLUSION: In this nonrandomized study of previously untreated patients with multiple myeloma, the addition of interferon in a dose of 3 mu/m2/day to dexamethasone achieved results similar to those with dexamethasone alone. PMID- 8402481 TI - Measurement and prediction of the short-term response of soft tissue sarcomas to chemotherapy. AB - BACKGROUND: Preoperative chemotherapy has been advocated for the treatment of soft tissue sarcomas, yet there is little information about how these tumors respond pathologically to therapy or whether tumor response can be predicted from a pretreatment biopsy. METHODS: Biopsy was done of 25 intermediate- or high-grade soft tissue sarcomas before they were treated with three cycles of doxorubicin and cisplatin and resected. The authors compared the pathologic features of the treated tumors with clinical and radiologic evidence of response to identify the pathologic features that best reflected chemotherapeutic effect. They analyzed the pretreatment biopsy specimens by light microscopic study and flow cytometry to identify parameters that predict short-term response to chemotherapy. RESULTS: In the treated tumors, the clearest indicator of early chemotherapeutic effect was the percentage of the resected mass composed of viable neoplasm. Eleven of 25 resection specimens contained less than 15% viable neoplasm (patients with pathologic response); 9 of these had 5% or less. Flow cytometric estimates of the proliferative rate in the initial biopsy specimen predicted early chemotherapeutic effect; 7 of 10 tumors with S-phase fractions (SPF) greater than 6% responded pathologically, whereas only 3 of 12 tumors with lower SPF responded (P = 0.041). Initial tumor grade, cell type, percent tumor necrosis, mitotic rate, cellularity, and ploidy did not predict chemotherapy response. CONCLUSION: These results indicate that soft tissue sarcomas often respond dramatically to chemotherapy, that the amount of residual viable sarcoma is an indicator of short term effect, and that flow cytometric estimates of cell proliferation predict early response to chemotherapy. PMID- 8402482 TI - Prognostic, quantitative histopathologic variables in lobular carcinoma of the breast. AB - BACKGROUND: A retrospective investigation of 53 consecutively treated patients with operable lobular carcinoma of the breast, with a median follow-up of 6.6 years, was performed to examine the prognostic value of quantitative histopathologic parameters. METHODS: The measurements were performed in routinely processed histologic sections using a simple, unbiased technique for the estimation of the three-dimensional mean nuclear volume (vv(nuc)). In addition, quantitative estimates were obtained of the mitotic index (MI), the nuclear index (NI), the nuclear volume fraction (Vv(nuc/tis)), and the mean nuclear profile area (aH(nuc)). RESULTS: Estimates of vv(nuc), MI, NI, and Vv(nuc/tis) were of significant or marginally significant prognostic value in univariate analyses, whereas no prognostic significance was attributed to estimates of aH(nuc). In multivariate Cox analyses, the clinical stage of disease, vv(nuc), MI, and NI were of significant independent, prognostic value. On the basis of the multivariate analyses, a prognostic index with highly distinguishing capacity between prognostically poor and favorable cases was constructed. CONCLUSION: Quantitative histopathologic variables are of value for objective grading of malignancy in lobular carcinomas. The new parameter--estimates of the mean nuclear volume--is highly reproducible and suitable for routine use. However, larger and prospective studies are needed to establish the true value of the quantitative histopathologic variables in the clinical management of patients with breast cancer. PMID- 8402483 TI - Communication between patients with breast cancer and health care providers. Determinants and implications. AB - BACKGROUND: This study evaluated the perceptions of patients with breast cancer of their medical interactions with providers. The determinants and psychological consequences of communication problems also were examined. METHODS: Ninety-seven patients with Stage I or II breast cancer completed a set of validated questionnaires before initiating postoperative therapy. Data on psychological distress were collected at baseline and 3-month follow-up, and multivariate models were fit to explain the relationship between pretreatment communication problems and subsequent psychological distress. Data on clinical variables were abstracted from medical records. RESULTS: A substantial proportion of patients (84%) reported difficulties communicating with the medical team. Communication problems were more common among patients who were less optimistic about their disease and had less assertive coping styles. Patient-reported communication problems were associated with increased anxiety, depression anger, and confusion at the 3-month follow-up. The association between communication problems and mood disturbance remained significant, although small, after adjusting for baseline mood disturbance, demographic, clinical, and coping style variables. CONCLUSION: Interventions that enhance communication between patients with breast cancer and their providers may improve patients' psychological adjustment to treatment. Conversely, interventions that lower distress and modify coping style may enhance communication. PMID- 8402484 TI - Transient increases of growth fraction during fractionated radiation therapy for cervical carcinoma. Ki-67 and PC10 immunostaining. AB - BACKGROUND: The Ki-67 and PC10 indexes are consistent with the growth and S-phase fractions, respectively. METHODS: Eighty-three biopsy specimens from 18 patients with cervical squamous cell carcinoma were examined before and during fractionated radiation therapy (RT), using an immunohistochemical method. RESULTS: The 18 patients were divided into two groups according to the Ki-67 index before RT; those with values of less than 42% (Group A; n = 11) and those with values of 42% or more (Group B; n = 7). The Ki-67 index in Group A increased approximately twofold at radiation doses of 5.4-9 Gy compared with values before RT. The Ki-67 index in Group B, which increased slightly at doses of 5.4-9 Gy, decreased by approximately one-fifth at a dose of 27 Gy, compared with the value before RT. The PC10 indexes in both groups showed little change to doses of 5.4-9 Gy. The mitotic indexes were less than 1% before RT and decreased to a minimum with doses of 18-27 Gy. There was no correlation between the Ki-67 and PC10 indexes before or during RT. CONCLUSION: In cancer tissues with a low growth fraction before RT, radiation doses of 5.4-9 Gy produced increases in the numbers of cycling cancer cells in the G1 or G2 phases. This transient increase possibly was attributable to the combined effects of the recruitment phenomenon and G1 or G2 block. PMID- 8402485 TI - Radiation therapy for prostate cancer. Long-term results and implications for future advances. AB - BACKGROUND: There are surprisingly few studies that concurrently report the 10 year overall survival (OS) rate, 10-year adjusted survival (AS) rate, 10-year recurrence free (NED) survival rate, and 10-year local tumor control (LC) rate using external beam radiation therapy (RT) for Stages A2, B, and C prostate cancer. A simultaneous analysis of OS, AS, NED survival, and LC rates is useful in terms of establishing priorities for future research efforts. METHODS: Actuarial LC, NED survival, AS, and OS rates were analyzed for 289 patients with Stages A2, B, and C prostate cancer treated with RT between 1975 and 1990 in the Department of Therapeutic Radiology, Yale University School of Medicine. RESULTS: The 10-year LC, NED survival, AS, and OS rates for 168 patients with Stage A2 and B disease were 88% (standard error [SE], 5%), 50% (SE, 10%), 70% (SE, 7%), and 38% (SE, 7%), respectively. The 10-year LC, NED survival, AS, and OS rates for 121 patients with Stage C disease were 82% (SE, 6%), 41% (SE, 6%), 25% (SE, 7%), and 21% (SE, 6%), respectively. CONCLUSION: External beam radiation provides excellent local clinical tumor control for early prostate cancer. However, 60% of all prostate cancer deaths of patients with Stage A2 and B disease and 76% of all prostate cancer deaths of patients with Stage C disease were attributable to metastatic disease without clinical local failure. The major obstacle in improving the death rate associated with Stages A2, B, and C prostate cancer remains the inability to control metastatic disease; research efforts must concentrate on overcoming this problem. PMID- 8402486 TI - Erythropoietin production. A potential marker for interleukin-2/interferon responsive tumors. AB - BACKGROUND: Interleukin-2 (IL-2) recently was approved by the Food and Drug Administration for the treatment of renal cell cancer. It is effective in a small minority of patients, but no markers identify individuals likely to respond to treatment. METHODS: Two polycythemic patients with erythropoietin-producing renal cell cancer and three other polycythemic patients with renal cell cancer were treated with the combination of IL-2 and alpha-interferon (alpha-IFN). RESULTS: All five patients achieved a partial or complete remission. In both patients in which it was measured, the erythropoietin level decreased significantly with treatment, and the polycythemia resolved in all patients. Hypothyroidism developed in two patients, and transient hyperthyroidism developed in another. CONCLUSION: These results contrast with those achieved with IL-2 alone or in combination with lymphokine-activated killer cells, for which a 15% response rate was seen in patients with renal cell cancer and polycythemia. Although less than 5% of renal cell tumors produce erythropoietin, its production may identify a subset of individuals with renal cell cancer responsive to IL-2 and alpha-IFN. PMID- 8402487 TI - Primary carcinoid of the kidney. Immunohistochemical and ultrastructural studies of five patients. AB - BACKGROUND: Primary carcinoid tumor of the kidney is rare; only 18 patients have been reported. Because of the rarity of the lesion, its prognosis and clinicopathologic features are not well characterized, and its histogenesis is unknown. METHODS: Clinicopathologic, immunohistochemical, and ultrastructural studies were performed on five patients with primary carcinoid tumor of the kidney. RESULTS: Age of the patients ranged from 42 to 63 years, with a mean of 53.2 years. Only one patient had a carcinoid syndrome. Three were men, and two were women. In each patient, the tumor was solitary and confined to the kidney. All tumors had histologic features identical to those of carcinoid tumors affecting other sites. The cytoplasmic granules of the tumor cells were reactive with Grimelius stain in all patients. All tumors also were immunoreactive for keratin, neuron-specific enolase, and chromogranin, but only two patients had tumors that reacted for serotonin, pancreatic polypeptide, and vasoactive intestinal polypeptide. Ultrastructural studies showed numerous intracytoplasmic neuroendocrine granules in four tumors. One patient had Robson Stage I cancer, and two each had Stage II and IV cancer. The two patients with Stage IV tumor died of disease at 36 months and 28 months after diagnosis; one patient with Stage II carcinoid is alive without clinical evidence of residual disease 8 years after control of metastases; the other patient with Stage II and the patient with Stage I cancer are alive and well without evidence of disease at 36 months, and 2 months, respectively. CONCLUSION: Tumor stage at presentation appears to be the most important factor in determining outcome. The number of mitoses and degree of cellular atypia appear to be important pathologic parameters in predicting prognosis, despite the limited number of patients reported in this report. PMID- 8402488 TI - Prognostic value of karyometric and clinical characteristics in renal cell carcinoma. Quantitative assessment of tumor heterogeneity. AB - BACKGROUND: The variation in tumor cell differentiation within one renal cell carcinoma, also termed tumor heterogeneity, renders visual tumor grading of these carcinomas difficult. Karyometric analysis enables description of nuclear characteristics of multiple tumor areas. Hence, karyometric analysis can be used to quantify tumor heterogeneity and thus may aid in a more objective grading of renal cell carcinoma. METHODS: In 121 patients with renal cell carcinoma (tumors in International Union Against Cancer [UICC] stages I [5 cases], II [23 cases], III [33 cases], and IV [60 cases]), clinical and karyometric features were studied to obtain routinely applicable prognostic factors. Several parts of the tumor were analyzed to obtain a measure of tumor heterogeneity. Univariate and multivariate Cox regression analyses were used to determine the predictive value of karyometric features independent of tumor stage and other clinical characteristics. RESULTS: The Cox univariate regression analysis showed correlation of several clinical and karyometric characteristics with survival. Of the clinical characteristics, TNM stage, tumor size, weight reduction, and performance status were significantly associated with survival. The karyometric features, especially those measurements associated with tumor heterogeneity (e.g. differences in nuclear size or chromatin texture between tumor subpopulations) were of value in predicting prognosis. In the Cox multivariate regression analysis, the Robson and UICC stages proved to be the most powerful predictors of survival (P < 0.0001). Of the clinical features, weight reduction and performance score were the only characteristics offering additional information regarding tumor stage (P < 0.0001). From the karyometric analysis quantification of anisokaryosis in the tumor at time of diagnosis offered additional prognostic information. Moreover, the differences of karyometric features within the tumor presumably associated with tumor heterogeneity correlated with survival. Using the features from the multivariate analysis, prognostic groups could be defined. CONCLUSION: We conclude that karyometric analysis offers a useful means for quantifying tumor heterogeneity. Multivariate Cox analysis revealed additional value of a grading system based on karyometric analysis to tumor stage. Karyometric analysis can be a useful tool for stratification of patient populations. PMID- 8402489 TI - Novel eosinophilic intracytoplasmic inclusions in a meningioma. AB - BACKGROUND: Two forms of inclusion, extracellular and intracytoplasmic, have been reported in meningiomas. The authors recently encountered a case of meningothelial meningioma with unusual round eosinophilic intracytoplasmic inclusions. METHODS: Ultrastructural and immunohistochemical studies were performed on the surgically removed tissue. RESULTS: Ultrastructurally, the inclusions were found in the cytoplasm and processes of the tumor cells. Each inclusion consisted of a dense central mass of filaments with a looser, less dense periphery of radiating filaments. At the periphery, crystalloid lamellar structures and granules were intermingled frequently. No limiting membrane was observed. Immunohistochemically, the inclusions were unreactive for five classes of intermediate filaments, actin, ubiquitin, and amyloid. CONCLUSION: The intracytoplasmic inclusions were ultrastructurally distinct from those reported previously. The molecular aspect of these novel eosinophilic intracytoplasmic inclusions remained uncertain. PMID- 8402490 TI - Prognostic importance of histologic grading in papillary thyroid carcinoma. AB - BACKGROUND: The influence of various clinicopathologic features on survival of patients with papillary thyroid carcinoma has been examined. In particular, histologic grade, which was obtained by a combined assessment of nuclear atypia, tumor necrosis, and vascular invasion (VAN score) was studied. According to this, histologic grade is independent of growth pattern and differentiation of the tumor cells. METHODS: One hundred seventy-three consecutive patients were studied retrospectively, and most were treated with total/near-total thyroidectomy. Univariate (life-table) and multivariate (Cox regression) analyses of thyroid cancer deaths were performed. RESULTS: Histologic grade showed a highly significant prognostic impact. It was possible to discriminate between low-grade (Grade 1) and high-grade (Grade 2) tumors, occurring in 79% and 21% of the patients, respectively. The risk of thyroid cancer deaths was found to be considerably higher in the latter group of patients. When histologic grade was combined with sex and age, a highly significant prognostic score (the SAG score) was obtained. CONCLUSION: Histologic grade based on nuclear atypia, tumor necrosis, and vascular invasion is a strong and independent prognostic factor. When combined with patient sex and age, the final SAG score and the pTNM-age classification were both significant and should be considered in prognostic stratification of these patients. PMID- 8402491 TI - Engraftment of human non-Hodgkin lymphomas in mice with severe combined immunodeficiency. AB - BACKGROUND: Malignant non-Hodgkin lymphoma (NHL) is one of the most difficult neoplasms to transplant into nude mice. Mice with severe combined immunodeficiency (SCID) accept various human cancers much more efficiently than do nude mice. The authors investigated whether SCID mice could be used as convenient hosts in which to grow human NHL in vivo. METHODS: Fifty NHL specimens were engrafted into SCID mice. The original specimens and the tumors that developed in SCID mice were studied immunohistologically and by Southern blot analysis to clarify their clonal identity and to determine if they were Epstein Barr virus (EBV)-transformed B cell proliferations. RESULTS: SCID tumors developed from 23 of 50 NHL specimens. Ten tumors were identical immunophenotypically and, partly, genotypically to the original NHL, showing that the original NHL grew in the SCID mice. B-cell NHL rather than T-cell NHL and high-grade rather than low-grade malignancy groups were much more easily heterotransplanted. Most of the heterotransplanted NHL were maintained by successive transplantation. In two other SCID tumors, the original NHL clones and a newly developed B-cell clone coexisted. The remaining 11 SCID tumors were composed of newly developed clones. The latter 13 tumors were shown to be human cells of B-cell lineage bearing EBV latent proteins--latent membrane protein 1 and EB nuclear antigen 2--suggesting that they originated from EBV-infected B cells that were present in the original tumor tissues. CONCLUSION: SCID mice accept human NHL far more efficiently than do nude mice. However, frequent occurrence of spontaneous EBV-associated B-cell proliferation must be kept in mind. PMID- 8402492 TI - Dose-ranging antiemetic evaluation of the serotonin antagonist RG 12915 in patients receiving anticancer chemotherapy. AB - BACKGROUND: RG 12915 is a potent, selective 5-HT3 receptor antagonist with a biologic half-life of 11-20 hours. RG 12915 prevents cisplatin-induced emesis in ferrets at doses of 0.03 mg/kg. Animal toxicology studies permitted safe testing in humans at doses of up to 2.0 mg/kg. This dose-ranging trial of intravenous RG 12915 was performed to determine the optimal dosage and adverse effects and to observe for antiemetic effects in patients receiving anticancer chemotherapy. METHODS: Twenty-six patients receiving chemotherapy likely to cause vomiting received a single intravenous dose of RG 12915 at a rate of 3 ml/minute beginning 60 minutes before chemotherapy. Four dose levels were explored: 0.25, 0.50, 1.0, and 2.0 mg/kg. RESULTS: No dose-limiting toxicities were observed. All adverse effects were mild and transient and included discomfort at the infusion site, hyperglycemia, headache, serum aspartate transaminase (AST) and alanine transaminase (ALT) elevations, and sedation. Antiemetic efficacy was seen in patients receiving cisplatin at doses of greater than or equal to 100 mg/m2. CONCLUSION: RG 12915 can be administered safely at the dose levels explored. Single intravenous doses of RG 12915 prevented or lessened emesis caused by chemotherapy, including cisplatin given at doses of greater than or equal to 100 mg/m2. RG 12915 warrants further testing. Of the doses tested, the 2.0 mg/kg dose is the most appropriate for further exploration. PMID- 8402493 TI - Sibling adult T-cell leukemia/lymphoma and clustering of human T-cell lymphotropic virus type I infection in a Jamaican family. AB - BACKGROUND: Human T-cell lymphotropic virus type I (HTLV-I) infection is endemic in Jamaica, with an estimated crude seroprevalence of 5%. Adult T-cell lymphoma/leukemia (ATL), a disease caused by HTLV-I, has an incidence of 1 2/100,000 in the Jamaican population. Familial ATL has not previously been reported from Jamaica. METHODS: Hospital records and histologic specimens of the two cases were reviewed. HTLV-I infection was confirmed by antibody testing and by polymerase chain reaction on paraffin-embedded tissue, where serum was unavailable. Family members were identified by the patients' parents. After giving informed consent, family members were asked to complete an interviewer administered questionnaire and to agree to phlebotomy. RESULTS: ATL developed 10 years apart in two siblings from a Jamaican family at age 16 and 24 years. A study of 19 members of their extended family, including both parents, 2 grandparents, and 3 siblings, revealed an overall HTLV-I seroprevalence of 17%. This compared with 75% among parents and siblings living in the same household as the patients. HTLV-I antibody-positive (HTLV-I-positive) and negative family members had similar mean age. Three of 3 HTLV-I-positive subjects were breast fed, compared with 10 of 15 HTLV-I-negative subjects. Intravenous drug abuse, sex with prostitutes, homosexuality, and blood transfusion were not reported. The mean number of sexual partners were similar. Both parents, who were antibody positive, had polylobated atypical lymphocytes in their peripheral blood. CONCLUSION: The HTLV-I antibody seroprevalence is greater in the family than in the general population, consistent with the modes of transmission. The antibody seronegativity of both grandmothers suggests sexual transmission between parents. The development of ATL at age 16 and 24 years is consistent with maternal-infant transmission and a long latent period, as reported by other authors. PMID- 8402494 TI - L-tryptophan-induced eosinophilia-myalgia syndrome associated with primary cutaneous malignant fibrous histiocytoma and extraabdominal desmoid tumor. AB - A 57-year-old woman with L-tryptophan-induced eosinophilia-myalgia syndrome, 23 months after the onset of symptoms, experienced development of a parasternal malignant fibrous histiocytoma on previously scleroderma-like skin areas and, almost concurrently, an extraabdominal desmoid tumor of the left arm muscle fascia. The malignant fibrous histiocytoma was treated by surgical excision without sign of recurrence or metastasis 19 months later. Radiation therapy was performed on the extraabdominal desmoid tumor. We suspect that these two connective tissue tumors in this patient were related to the exposure to contaminated L-tryptophan, which interfered with connective tissue metabolism. PMID- 8402495 TI - Diagnostic problems of second primary malignancies detected by fine-needle aspiration cytology. AB - BACKGROUND: The diagnosis of second malignant tumors, the incidence of which is increased because of the increased duration of survival for many tumor types in patients treated for cancer, represents one of the most important aspects of application of fine-needle aspiration (FNA). METHODS: Forty patients who had been treated for a malignant tumor and who developed a second primary tumor detected by FNA and subsequently confirmed by histologic examination were studied. The primary tumors were malignant lymphoma (15 cases); breast carcinoma (10 cases); cutaneous melanoma (3 cases); carcinoma of the larynx and of the uterine cervix (2 cases each); and dysgerminoma of the ovary, chronic myeloid leukemia, and carcinoma of the nose, floor of the mouth, tonsil, anus, kidney, and skin (one case each). The interval between the diagnosis of the first and second malignant tumors ranged from 1 month to 22 years. RESULTS: Carcinoma of the lung, mainly represented by adenocarcinoma, was the most frequent second malignant tumor that occurred in this series (15 cases). It was followed by carcinoma of the breast (5 cases); soft tissue sarcoma (3 cases); and carcinoma of the liver, kidneys, ovaries, thyroid gland (2 cases each), parotid gland, and skin. Five cases of non Hodgkin lymphoma and one of Hodgkin disease developed in patients with a previous carcinoma at various sites. Four non-Hodgkin lymphomas occurred as the second primary malignant tumor in patients treated with radiation therapy for a mammary carcinoma. One case of multiple myeloma occurred. CONCLUSION: The accuracy of FNA in diagnosing second primary tumors (100%) and in classifying them by histologic type (62.5%) was demonstrated. PMID- 8402496 TI - Sweet syndrome in patients with solid tumors. AB - BACKGROUND: Sweet syndrome (acute febrile neutrophilic dermatosis) may occur as a cutaneous paraneoplastic syndrome. This condition has been associated with hematologic malignancies and, to a lesser extent, with solid tumors. METHODS: The authors report two patients with malignancy-associated Sweet syndrome: a 66-year old man in whom the onset of Sweet syndrome preceded the diagnosis of an adenocarcinoma of unknown primary by 3 months and a 69-year-old woman in whom a workup after the appearance of Sweet syndrome skin lesions revealed an unsuspected recurrent squamous cell carcinoma of the larynx. The authors review the reports of the other 39 patients with solid tumor-associated Sweet syndrome that have been published in the world literature. RESULTS: The most common malignancies were carcinomas of the genitourinary organs (37%), breast (23%), and gastrointestinal tract (17%). Typical clinical features and laboratory findings in these patients included tender erythematous plaques located on the upper extremities (97%); elevated erythrocyte sedimentation rate (95%); anemia (83%); fever (79%); and neutrophilia (60%). The symptoms and lesions of Sweet syndrome resolved after treatment with corticosteroids, potassium iodide, or colchicine. Sweet syndrome preceded the initial diagnosis of cancer or the detection of asymptomatic metastatic, persistent, or recurrent tumor, or a hematologic malignancy (in an individual with a previously diagnosed solid tumor) in 61% of the patients. In the other 39% of patients, diagnosis of Sweet syndrome followed the development of a solid tumor. CONCLUSION: The search for a neoplasm of the genitourinary organs and breast cancer in women and a gastrointestinal tract carcinoma in men should be emphasized in the evaluation for a solid tumor in patients with Sweet syndrome without a prior diagnosis of malignancy. PMID- 8402497 TI - Parenteral nutrition associated with increased infection rate in children with cancer. AB - BACKGROUND: Recent meta-analyses of published controlled studies concluded that adult patients with cancer randomly assigned to receive parenteral nutrition had higher rates of infectious complications than control subjects. METHODS: The infection risk associated with parenteral nutrition was assessed in 310 pediatric patients with cancer. These patients had central venous access devices (CVAD), Hickman/Broviac (H/B) catheters, or implantable subcutaneous ports in place for the delivery of chemotherapy and supportive care. RESULTS: The median duration of CVAD placement was 363 days; a total of 450 patient years (i.e., the sum of the total years of catheters experienced from all patients studied) were examined. Overall, the infection rate was 0.06 infections/100 days. During the period of parenteral nutrition administration, the rate increased to 0.5 infections/100 days. Among patients who received parenteral nutrition, there were no significant differences in any clinical parameter between the patients who developed an infection and those who did not. When evaluating the entire study population, infection was more likely to occur in patients who had acute nonlymphocytic leukemia (P < 0.01) or H/B catheters (P < 0.01), or who received parenteral nutrition (P < 0.02); there was no relationship between infection and catheter duration, days hospitalized, or days neutropenic (absolute neutrophil count < 0.5 x 10(9)/l). Only CVAD type and parenteral nutrition retained significance in a multivariate Cox proportional hazards model. After adjustment for diagnosis and CVAD type, the risk of infection was 2.4-fold greater in patients given parenteral nutrition (95% confidence interval 1.5 to 3.9; P < 0.001). CONCLUSION: These data confirm that administration of parenteral nutrition is associated with an increased risk of infection in children who have CVAD in place for cancer therapy. PMID- 8402498 TI - Comparison of cardiac function tests after anthracycline therapy in childhood. Implications for screening. AB - BACKGROUND: Numerous methods for evaluating cardiac function after cardiotoxic therapy have been suggested. A practical algorithm for screening a large patient population is needed. METHODS: Seventy-three patients (median age, 15.3 years [range, 9-29 years) who received anthracyclines (median dose, 300 mg/m2 [range, 50-750 mg/m2) for a childhood malignancy underwent a battery of commonly used tests of cardiac function: (1) echocardiographic shortening fraction (ESF); (2) resting and exercise radionuclide scan (multiple gaited acquisition [MUGA]); (3) electrocardiogram (ECG); (4) 24-hour Holter monitor, and (5) questionnaire concerning exercise tolerance. RESULTS: Patients with an abnormal resting ejection fraction on MUGA were more likely to have an abnormal ESF (P = 0.023), prolongation of the QTc interval (P = 0.014), and complaints of "difficulty" with exercise (P = 0.04). An abnormal results for a resting study was not predictive of an abnormal MUGA exercise response. There was no association between the presence of significant dysrhythmia on Holter monitor and any resting test. An algorithm was developed using a normal ESF and QTc interval, and a negative history of exercise intolerance to identify patients who need not undergo a MUGA. With the use of such an algorithm, 29 of the 73 patients would not have undergone the MUGA, only 2 of whom had an abnormal resting ejection fraction. CONCLUSION: Patients at risk for cardiac damage after cancer therapy who have normal ESF and QTc interval results and no history of exercise intolerance are unlikely to have abnormal MUGA results. Significant dysrhythmia may be present in the absence of other abnormalities. PMID- 8402499 TI - Neurofibromatosis type 1 and childhood cancer. AB - BACKGROUND: Patients with neurofibromatosis type 1 (NF1) are prone to develop malignancy, particularly malignant schwannoma and glioma in adults. METHODS: To assess the risk for childhood malignancy in NF1, 26,084 patients with cancer younger than 15 years of age registered from 1969 to 1989 in the Japan Children's Cancer Registry were reviewed. The incidence of NF1 in each type of cancer was compared with that in the Japanese population. RESULTS: Fifty-six children with cancer had NF1 in the national registry. The incidence of NF1 (0.21%) was 6.45 times that of the expected estimated rate of 1 per 3000 in the Japanese population. These tumors tended to be type and site specific. The NF1 incidence was extremely high in optical nerve glioma (12.5%), other central nervous system gliomas (0.9%), and malignant schwannoma (31.4%). For nonneural tumors, NF1 incidence was increased in rhabdomyosarcoma (1.36%), particularly those in urogenital organs, and in myelogenous leukemia (0.27%). Epithelial carcinomas were not observed in the group of patients with NF1. CONCLUSION: The risk for glioma and malignant schwannoma increases in children and adults with NF1. Moreover, the risk for two childhood malignancies, myelogenous leukemia and rhabdomyosarcoma, increases in children with NF1. The NF1 gene seems to increase the risk not only for neural tumors but also for some non-neural tumors in an age specific, organ-dependent pattern of carcinogenesis. PMID- 8402500 TI - Preradiation chemotherapy in advanced medulloblastoma. A Pediatric Oncology Group pilot study. AB - BACKGROUND: Children diagnosed with medulloblastoma whose tumor involves the brain stem or has spread through the cerebrospinal fluid pathways to other areas of the brain or spinal cord have a poor prognosis despite therapy with surgery, craniospinal irradiation (CSI), and chemotherapy. Preradiation chemotherapy may improve the outlook for these patients. METHODS: To further study the role and feasibility of preradiation chemotherapy, children between the ages of 4 and 21 years diagnosed with advanced medulloblastoma and measurable disease were enrolled in the Pediatric Oncology Group 8695 study. Patients were treated with a 9-week course of vincristine, cisplatin, and cyclophosphamide followed by CSI. Imaging films were reviewed centrally for eligibility and response to chemotherapy. Toxicity to chemotherapy and radiation as well as delays and modifications in subsequent CSI were recorded. RESULTS: Thirteen of 30 fully evaluable patients achieved complete or partial response (43%) to chemotherapy. Toxicity was mostly fever and neutropenia after cyclophosphamide, which is predictable and tolerable. Radiation therapy was delivered in full doses and volumes in most patients but was delayed in its start in most patients. Central review of films revealed frequent use of different imaging modalities at baseline and after therapy, making accurate assessment of tumor response difficult. CONCLUSION: Preradiation chemotherapy with vincristine, cisplatin, and cyclophosphamide is active in patients with advanced medulloblastoma but should be modified to minimize the risk of progressive disease while on therapy and to avoid delays in starting radiation therapy. Consistent use of the same neuroimaging modality is essential in documenting response. PMID- 8402501 TI - Combined determination of N-myc oncogene amplification and DNA ploidy in neuroblastoma. Complementary prognostic indicators. AB - BACKGROUND: N-myc gene amplification is a well-established prognostic indicator in neuroblastoma. Flow cytometric analysis of nuclear DNA content has shown that an abnormal nuclear DNA content in neuroblastoma is associated with a better prognosis. Because some patients with N-myc unamplified tumors have a poor prognosis, factors other than N-myc amplification may play a role in determining the clinical behavior of neuroblastoma. In the current study, the authors correlated N-myc gene amplification and flow cytometric nuclear DNA content with respect to prognosis. METHODS: Forty-one patients with neuroblastoma, including 15 screened patients, served as subjects. The copy number of the N-myc gene was determined by Southern blot analysis. DNA ploidy analysis was done on nuclei isolated from formalin-fixed, paraffin-embedded blocks. RESULTS: Of 40 specimens of neuroblastoma, 7 involved tumors containing amplification of the N-myc gene and 33 did not; 13 specimens showed DNA diploidy, and 27 showed DNA aneuploidy (including 4 with DNA tetraploidy). The Kaplan-Meier survival analysis indicated a significantly better prognosis in patients with unamplified N-myc tumors compared with those with N-myc amplified tumors (87.3% versus 28.6%, P < 0.05) and in patients with DNA aneuploid tumors compared with those with DNA diploid tumors (96.3% versus 43.0%, P < 0.001). The difference in the survival of the two extreme combinations, (e.g., 25 with N-myc unamplified and DNA aneuploidy [4 tetraploidy] versus 5 with N-myc amplified and DNA diploidy) was more significant (96.0% versus 20.0%, P < 0.001) than any other combination. CONCLUSION: Evaluations of N-myc gene amplification and DNA ploidy are complementary, and the combined determination of these two factors may be one of the most powerful prognostic indicators in neuroblastoma. PMID- 8402502 TI - Aggressive chemotherapy, organ-preserving surgery, and high-dose-rate remote brachytherapy in the treatment of rhabdomyosarcoma in infants and young children. AB - BACKGROUND: External beam radiation therapy often is avoided in the treatment of rhabdomyosarcoma (RMS) in young children because of the long-term sequelae. Conventional brachytherapy can reduce these problems, but its use is limited in young children because of radiation exposure to parents and care-givers. This is the first reported use of high-dose-rate remote brachytherapy (HDR) to treat RMS in young children. METHODS: Seven young children with RMS were treated from January 1990 through September 1991 with multiagent chemotherapy, organ preserving surgery, and HDR. The primary tumor sites included the tongue, buccal mucosa, chest wall, vagina, and clitoris. A minimum peripheral dose of 36 Gy HDR was administered in 12 fractions (twice a day) at 3 Gy per fraction for a period of 3 days. The treatment was given on an outpatient basis without requiring prolonged patient sedation or immobilization. Each treatment lasted 2-5 minutes. RESULTS: All seven children are alive and without evidence of tumor with a median follow-up of 30 months (range, 18-35 months) from diagnosis. The treatments have been reasonably well tolerated with some acute skin toxicity. There has been relatively good organ growth and function during this short follow-up period. CONCLUSION: The use of HDR radiation in these patients eliminated radiation exposure to care-givers and permitted constant nursing care and interaction among the parents, nursing personnel, and child. Treatments can be given on an outpatient basis, without requiring prolonged patient sedation or immobilization. Local control of tumor with preservation of organ function was achieved. HDR in young children should be restricted to controlled clinical trials until long-term morbidity and efficacy results are obtained from pilot studies. PMID- 8402503 TI - Cancer in Peutz-Jeghers syndrome. AB - BACKGROUND: Peutz-Jeghers (P-J) syndrome has been found to be associated with an increased risk of malignant neoplasia. METHODS: The authors reviewed the clinical courses of eight patients with P-J syndrome (four male and four female patients, ranging in age from 8 to 55 years), who had been followed up for as long as 12 years. The diagnosis of this syndrome was based on the evidence of characteristic mucocutaneous pigmentations and gastrointestinal hamartomatous polyposis. RESULTS: Four cases of malignant neoplasm among the eight patients were found. In a 25-year-old woman, an extremely well-differentiated adenocarcinoma of the uterus cervix was found at the initial examination. A gastric cancer in a 32-year old woman, duodenal cancers in a 43-year-old man, and a pancreatic cancer in a 60 year-old man also were identified 12, 10, and 5 years after the time of initial examinations, respectively. Two of these patients died of the effects of the tumor soon after. CONCLUSION: The authors' experience confirms a veritable malignant potency in P-J syndrome and suggests that an intensive follow-up of gastrointestinal and extra-gastrointestinal sites is needed in patients with this syndrome. PMID- 8402504 TI - Posttransplantation lymphoproliferative disorders treated with cyclophosphamide doxorubicin-vincristine-prednisone chemotherapy. AB - BACKGROUND: Posttransplantation lymphoproliferative disorders after solid organ transplantation are a serious complication occurring in 1-10% of patients. Different therapies have been used, but the optimal treatment is unknown. There is relatively little information in the literature on the experience with cytotoxic chemotherapy. METHODS: The disease stage of patients with biopsy documented posttransplantation lymphoproliferative was determined with standard methods to establish the extent of the disease. Patients in whom the disease failed to regress after initial management, which included reduction in immunosuppression, were treated with a combination chemotherapy regimen consisting of six cycles of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). Response to therapy was determined by following previously defined sites of disease with appropriate tests. Patients were maintained on a reduced dose of immunosuppressants. RESULTS: In the four patients studied, lymphoproliferative disorders developed after heart (three cases) or lung (one case) transplantation, which did not regress after immunosuppression was reduced. All four experienced a complete remission with CHOP chemotherapy, which continued at 3, 13+, 20 and 30+ months after completion of treatment. One patient died of sepsis after completing therapy at a point when his leukocyte count was normal, and no evidence of posttransplantation lymphoproliferative disorder was found at autopsy. A second patient died of liver failure with no clinical evidence of lymphoproliferative disorder. CONCLUSION: Although this is a small series, it demonstrates that patients with posttransplantation lymphoproliferative disorders may respond to cytotoxic chemotherapy. The duration of response is undetermined. PMID- 8402505 TI - The request for assistance in dying. The need for psychiatric consultation. AB - BACKGROUND: Public initiatives and legislative proposals have increased the likelihood that some states will legalize euthanasia and assisted suicide as a means of ending the suffering of patients with terminal illness. However, suggested safeguards that would guide physicians in such cases have not properly addressed the need to evaluate psychosocial factors that could motivate patients' requests for premature death. METHODS: Four cases of patients with cancer who expressed a wish to end their lives prematurely are described. These cases were evaluated with regard to mental and emotional functioning. RESULTS: Pain and suffering, organic mental disease, depression, and personality issues play significant roles in patients' requests for assistance in dying. CONCLUSION: Comprehensive psychosocial assessment is needed when evaluating requests for assistance in dying. This assessment may reveal hidden problems or conflicts that affect rational decision making, a prerequisite to informed consent for any procedure or intervention. PMID- 8402506 TI - Patient selection for clinical trials. Risks versus benefits and quality of life issues. AB - Although some clinician-scientists have expressed reservations regarding utilization of randomized treatment trials, the majority of clinical investigators involved with cancer believe that the well-designed, properly executed clinical trial is the optimal treatment approach if a patient is eligible. To conduct such clinical trials in an ethical fashion, stringent criteria for patient selection are of critical importance both to protect the patient's rights and to assure valid data on outcomes. These selection factors, both ethical and scientific, will be reviewed. Also, the benefits and potential risk factors for participation in the clinical trial that need to be reviewed as part of the informed consent process will be discussed. The cost/benefit ratio should favor participation in the trial, if the trial is properly designed. Quality of life issues, particularly toxicities resulting from one or more treatment alternatives in trial, are an important part of the foregoing analysis. In addition, quality of life endpoints are becoming more important in the planning of clinical treatment trials for cancer, and examples of these will be discussed. In an overall sense, the practical ethics of randomized clinical trials for investigators will be addressed from the standpoint of the impact on potential patient participants. PMID- 8402507 TI - Cancer therapy and the randomized clinical trial. Good medicine? AB - True improvements in the treatment of cancer--by the introduction of new drugs or novel drug combinations, new therapeutic modalities, or technologic improvements of old modalities--result in higher response rates and prolonged survival when compared with existing therapies. When a new treatment convincingly meets the test of improving survival rates or, at worst, improving patients' quality of life, it becomes the accepted standard of care if its side effects are acceptable and its cost is not prohibitive. Improved therapeutic results can be demonstrated only by clinical trials with an adequate numbers of patients, appropriate control subjects, and a sufficient duration of follow-up. Therapeutic breakthroughs are revolutionary advances in treatment, usually rapidly and dramatically obvious in comparison with historic controls; demonstration of benefit in these cases does not usually require randomized trials. Much more common, however, are new therapies that represent modest, incremental advances over existing treatment and that usually require randomized comparison trials to demonstrate convincingly statistically significant improvement. A randomized clinical trial should test an important hypothesis. It must be carefully designed to ensure that both groups of patients are comparable in terms of various prognostic variables and to minimize subtle sources of bias. An honest belief that both arms of the trial are a priori equal must be maintained. Meeting these criteria, the randomized clinical trial offers to the individual cancer patient treatment that should be at least equal to the best available nonexperimental therapy. This equates with Good Medicine. PMID- 8402508 TI - Ethics of clinical trials. Do they help the patient? AB - Ethical justification of a randomized clinical trial requires at the onset that the investigators be able to state an honest null hypothesis regarding the therapies to be compared. For example, in a randomized clinical trial with only two arms (A and B), it is necessary to state that there is no scientifically validated reason to predict that therapy A will prove to be either superior or inferior to therapy B. Moreover, there must be no therapy C known to be superior to A and B unless there is a cogent reason to reject therapy C; the population of research subjects will consist either of those in whom therapy C has been tried without success or individuals who are aware of therapy C and have rejected it for various reasons. Thus, theoretically, the patients enrolled in a randomized clinical trial are assured that they will receive one of the two best known therapies for their condition. At first glance, one might conclude that randomized clinical trials do, in general, help the patient. There are some features of at least some clinical trials, however, that may be detrimental to the interests of some patients. It commonly is said, for example, that patients in randomized clinical trials are deprived of the "good of personal care." The physician-investigator may be constrained by rigid protocol requirements from being responsive to the particular needs and desires of the patient. Additionally, the process of obtaining informed consent to randomized clinical trials may be preemptive: patients who might have chosen palliative care rather than chemotherapy may not be afforded adequate opportunity to make such a choice. Any randomized clinical trial should maximize the likelihood of helping the patient. PMID- 8402509 TI - Informed consent. The whole truth for patients? AB - Two misconceptions about informed consent concern the difference between the legal and moral justification for it, and the rationale for more rigorous consent for research than for "standard" therapy. Legally, informed consent for therapy is a risk-management tool that functions essentially as a release of liability. If the patient is informed of all expected or potential side-effects or toxicities of a treatment, he cannot sue because those side effects occur. The moral doctrine of informed consent is derived from a respect for the patient's autonomy as well as the patient's vulnerability. The physician's goal is not to minimize liability, but to help the patient make the best decision. These two goals are not necessarily incompatible, but they often lead to different attitudes toward informed consent and different decisions about what information needs to be shared with patients. If the goal is risk-management, then informed consent forms should be encyclopedic, providing the "whole truth" to patients. This would, however, not meet the moral goals of shared decision-making, because few patients could make sense of such data. Informed consent for research often leads to a different paradox. If a new therapy becomes available and a physician thinks it may benefit a patient, the physician may use it in an uncontrolled manner. This does not require the approval of an institutional review board and does not need the more rigorous approach to informed consent generally associated with research. In contrast, if the physician wants to evaluate the effectiveness of the same therapy carefully and intends to gather data to do so, he will need to describe his protocol, defend it before an institutional review board, and provide a more carefully written and closely scrutinized informed consent form. The paradox derives from the fact that the patient in the first, uncontrolled situation is inevitably at higher risk of harm than the patient in the second situation. The distinction between informed consent for treatment and informed consent for research is based on a distinction that is growing ever more cloudy and ever less relevant to the moral goals that informed consent seeks to achieve. Current standards for informed consent for research are, in many ways, counterintuitive and probably counter-productive. PMID- 8402510 TI - The ethical treatment of cancer. What is right for the patient? PMID- 8402511 TI - A large private university hospital system. The Johns Hopkins Oncology Center. AB - Clinical trials are a major commitment for a university-based comprehensive cancer center. In 1992, The Johns Hopkins Hospital registered 3508 new patients with cancer and, from this large population, 2880 patients were entered in clinical trials (many patients participated in more than one protocol). The Oncology Center, one of many departments at Johns Hopkins that conducts clinical research, participates in phase I and II new drug trials, phase III comparative studies, and, increasingly, in epidemiologic and prevention research. This calls for much broader participation by community hospitals and for many more patients who normally would not come to Johns Hopkins for their care. There are more than 100 protocols available from the Eastern Cooperative Oncology Group, but Johns Hopkins may participate in no more than 20 at any given time. Thus, every research facility must be selective about the trials in which it participates, given the finite number of hours, dollars, and resources available to carry out these programs. The institution provides safeguards to protect the interest of the patient. These include review and annual overseeing of the concept, design, and specifics of the proposed study. The pharmacy and nursing staff play an important role in control of chemotherapy distribution and use. Patients and physicians, however, must understand the questions the study is asking and agree that they are worth answering. There are problems in motivation; information; costs to the patient, hospital, insurers, and the physician; the concept of the placebo; and informed consent. Clinical research is the most ethical way to test drugs, radiation therapy, surgical procedures, or other new treatments. The clinical trial must meet rigorous criteria of design, conduct, and analysis. The patient must understand the issues and be a volunteer. We must make every effort to help patients and physicians get information about clinical trials and to participate if they choose. PMID- 8402512 TI - Impact of clinical trial protocols on patient care systems at the University of Texas M.D. Anderson Cancer Center. AB - Clinical trial protocols represent a significant component of the research program at the University of Texas M.D. Anderson Cancer Center. Currently, there are nearly 800 active protocols using more than 150 investigational drugs. Sponsors of these trials include the National Cancer Institute, numerous pharmaceutical companies, and a variety of other funding sources. Because of our large patient base, however, routine participation in cooperative groups is limited. The impact of clinical trial protocols on patient care systems is a growing concern that is addressed primarily by the Surveillance Committee at M.D. Anderson. Like many institutional review boards, this committee reviews all protocols and must grant approval before the activation of any clinical trial. The issues of medical ethics, including a risk-benefit assessment for research subjects, equitable patient selection, safeguards for economically disadvantaged populations, and the general requirements for informed consent, also fall under the purview of this board. For any comprehensive cancer center to succeed, it must have strong research programs that enhance treatment options for patients. The crisis in financing health care significantly threatens the viability of cancer research. Reimbursement for the related costs of individuals participating in clinical trials has become a battle ground for third-party payers. The resulting economic risk is shared by health care providers and patients. As a consequence, many cancer centers, including M.D. Anderson, have initiated an economic-impact analysis for protocols as a component of the review process. This analysis includes an estimation of the patient care costs compared to the funding allocated to support the research. Future research will include pharmacoeconomic analysis and other ways to relate the costs and outcomes of treatment protocols. In the interim, we must monitor reimbursement trends for patients enrolled in clinical trials closely and amend the informed consent process to include a discussion of potential economic risk or hardship to patients if insurance claims are denied. Cancer care providers, third-party payers, the federal government, and society at large must work together to address the funding of cancer research and patient care. PMID- 8402513 TI - The impact of clinical trial protocols on patient care in a community hospital. AB - Although clinical research is essential in cancer control, less than 5% of cancer cases in community hospitals are entered in clinical trials. Phase III studies comprise the bulk of clinical research at our community hospital. Potential research protocols are selected by the Community Clinical Oncology Program Therapeutic Investigations Committee for referral to affiliated institutional review boards. Selection criteria include assessments of the scientific validity, relevance, and importance of the research, the match between the proposed protocol and research interests and experience of the investigators, priorities of the research bases for patient accrual, and resource availability for conduct of the study. There are serious impediments to the performance of clinical trials in the multiethnic, multilingual urban community hospital setting. These include diminution of the potential patient pool for trial entry due to language barriers and other difficulties associated with patient accrual; escalating costs (insurance coverage from some carriers in not available for clinical trial participation, and some therapies may be excluded by third-party payers); underinsurance or lack of insurance in many segments of the potential patient population; declining financial support for clinical research and the variability of payments, with restrictions even for standard or conventional treatments; the widespread belief that investigational studies are too costly or are of unproven value; potential liabilities associated with withholding cancer therapy; the unresolved legal quagmire surrounding cancer care for patients just under 18 years of age; and the potential for scientific misconduct related to the performance of clinical trials. PMID- 8402514 TI - The impact of clinical trial protocols on patient care systems in a large city hospital. Access for the socially disadvantaged. AB - Some Americans suffer a higher cancer incidence and mortality than those in mainstream American society, and, in general, do not enjoy the same health status. Black Americans, for example, have higher cancer incidence and lower survival rates than do white Americans. To date, there is no known genetic basis to account for the disparities in cancer incidence and outcome between these races. Controlling for socioeconomic status greatly reduces, and sometimes nearly eliminates, the apparent contrast in cancer mortality and incidence between ethnic groups. Poverty clearly is associated with diminished access to health care, an increased incidence of cancer, and 10-15% lower 5-year survival rates. Diminished access often is manifested by low quality and inadequate continuity of health care, as well as insufficient access to methods of disease detection, diagnosis, treatment, and rehabilitation. Poor people tend to concentrate on day to-day survival, often feel hopeless and powerless, and may become socially isolated. It is more difficult to conduct cancer treatment trials in a population characterized by such dramatic socioeconomic and cultural differences. Lack of insurance and lack of compliance become trial-limiting issues. This paper examines what must be done to tear down the economic and cultural barriers to prevention, early detection, and treatment of cancer. PMID- 8402515 TI - The impact of clinical trial protocols on patient care systems. PMID- 8402516 TI - Reimbursement issues facing patients, providers, and payers. AB - Escalating costs of health care delivery and the current constraints imposed by the federal budget deficit seriously threaten to compromise patient care and innovative biomedical research. Recent third-party refusal to cover some patients treated in protocols has had considerable impact on trial research. In addition, reimbursement for conventional care sometimes has been refused if delivered as part of a study (e.g., MOPP therapy versus ABVD therapy) or for an indication that is not specifically cited on the Food and Drug Administration label. Who should cover the patient care costs of patients participating in clinical trials? One approach would have patients cover these costs themselves. A second approach is the reinstitution of patient care costs into research grants. A third possibility is that the pharmaceutical industry support patient care costs of clinical research. Historically, hospital expenses of patients participating in studies have been paid by health insurance policies. In the absence of a clinical trial, many patients would be treated with Food and Drug Administration-approved therapies despite a lack of substantial benefit. Such marginal treatments are compensated by third-party payers routinely. The current system is arbitrary and expensive, compromises research and development, and equates new treatment with no treatment. By refusing to reimburse the patient care costs of investigational therapy, third-party carriers are, in fact, making medical decisions. There is a growing and legitimate concern that the pace of clinical research will be impeded significantly at a time when many exciting developments will be ready for clinical trials. The molecular steps in carcinogenesis are being documented rapidly for common malignancies, such as colon cancer. Immunologic, biologic, and hormonal approaches, and emerging technologies, such as marrow transplant or antibody toxin conjugates, already are being studied in the clinic. Health policy legislation and decisions by the insurance industry have a direct impact on physicians, facilitating, or often impeding, the care physicians are able to provide. Who makes health policy decisions? Increasingly, these decisions are being made not by physicians, but by public health experts, economists, and, more recently, large industries grappling with the cost of providing insurance coverage (and its effects on competitive pricing in a world market). Therefore, physicians need to position themselves to influence the development of medical policy, particularly as it relates to the prevention, diagnosis, and treatment of patients with cancer. PMID- 8402517 TI - Psychosocial effects of cancer economics on patients and their families. AB - Cancer frequently follows an unpredictable course, with patients experiencing numerous disruptions in their lives. Concomitantly, the enormous financial burdens that accompany the onset and subsequent treatment of cancer become even more overwhelming. Regardless of socioeconomic status, almost all families confronted with cancer and its treatment will have financial problems. Poor people are more likely to be diagnosed with cancer when the disease is advanced and treatment options are significantly more limited. Limited access to medical care carries the additional risk of denied access to community resources, which often are made through referrals from the health care system. For the middle class family with insurance, as medication costs increase (whether they are covered or not), financial deprivations accumulate as out-of-pocket expenditures, because of required insurance deductibles and copayments and wages lost because of aspects of the treatment. Therefore, even those who are insured can be financially devastated by substantial gaps in coverage. A diagnosis of cancer compounds the struggle for survival and introduces new financial, physical, and psychologic demands. As a nation, we must ensure that cancer prevention, detection, treatment, and rehabilitation services are accessible and available to all who need them, regardless of their ability to pay. PMID- 8402518 TI - The impact of health-care reform on the cancer patient. AB - Reform of the United States' health-care system as we know it will require a reevaluation of the methods used in health-care delivery. Many changes are emerging in the health-care marketplace, and there is a prevailing trend toward universal coverage through managed care/competition. Global cost-control measures will become more prevalent, as we have seen already in the Clinton Administration's plan to become the sole and price-controlling customer for all childhood vaccines. Health-care delivery is evolving toward local care networks involving managed competition, a system in which patient groups will trade volume for price. Legal barriers presently include antitrust laws, which restrict collaboration among health providers, fraud and abuse laws, and tax considerations. Licensure and regulatory issues also may have an impact on reforms. The benefits most likely will include guaranteed health care, reduced health-care costs, and a better, more efficient quality of medical care. With these changes, however, there will be less freedom of choice in health care and a greater burden on certain sectors of the economy. PMID- 8402519 TI - Ethical and financial considerations in third-party support of investigational cancer therapies. AB - The appropriate use of and compensation for investigational cancer therapies raises a number of ethical, policy-related, and financial considerations. Ethical considerations include the use of therapeutic regimens for individuals versus controlled clinical trials, the extent of information necessary to indicate that a therapy may be safe and effective, and the informed consent of the patient. Financial concerns include the status of the therapy in the evaluation phase within the Food and Drug Administration, the policy of third-party payers, and the current state of medical knowledge. The policy-related considerations are those regarding the use of and payment for investigational therapies. PMID- 8402520 TI - Government support of cancer research and cancer care health benefits. AB - As the federal government struggles to meet ever-growing budgetary constraints, the need for research into the causes of and potential therapies for treating cancer demands the urgent attention of Congressional leaders. This will require innovative, cost-effective approaches to address the unique needs of cancer researchers, physicians, and patients. As a new member of the Senate Appropriations Committee and a member of the Labor, Health and Human Services, and Education Subcommittee, I will be in the forefront of this important debate; as a cancer survivor, I will approach this new assignment with a deep personal conviction. Key issues in the fight against cancer will focus on the way to make cancer screening available to all Americans, the National Cancer Institute Bypass Budget, and the process by which the Food and Drug Administration approves new therapies. PMID- 8402521 TI - The economic impact of therapy on cancer patients. PMID- 8402522 TI - 3D base: a geometrical data base system for the analysis and visualisation of 3D shapes obtained from parallel serial sections including three different geometrical representations. AB - In this paper we discuss a geometrical data base that includes three different geometrical representations of one and the same reconstructed 3D shape: the contour-pile, the voxel enumeration, and the triangulation of a surface. The data base is tailored for 3D shapes obtained from plan-parallel serial sections. It is explained how this geometrical data base is useful with the different processing approaches of a 3D shape, such as analysis and visualisation. Methods of conversion between the geometrical representations are discussed. Examples of the operation of the data base as it is embedded in a data base management system are given by illustrations of retrieval of geometrical information. PMID- 8402523 TI - 3D reconstruction of biological objects from sequential image planes--applied on cerebral cortex from cat. AB - A prism of cat cerebral cortex was reconstructed with a method for three dimensional (3D) representation of biological objects. A series of 918 semithin sections were digitized into an image analysis system. The images were aligned and analyzed, and a data base with the coordinates and a classification of the cells was created. The data base (i.e., the cortical prism) was visualized in a 3D graphic terminal, and parameters such as columnar and lamellar organization, clustering, and cell density were analyzed. A neuronal perikaryon and its neurites was reconstructed and shown together with the cortical prism. PMID- 8402524 TI - Segmentation and measurement based on 3D Voronoi diagram: application to confocal microscopy. AB - Computational geometry provides many solutions to imaging problems, especially for three-dimensional (3D) image compression, segmentation, and measurement. We present here a new method to partition volume data by Voronoi polyhedra structured in a graph environment. A dynamic construction of the 3D Voronoi diagram is proposed, using image information interactively. The process has been applied to segment and quantitate 3D biological data acquired with a confocal laser scanning microscope (CLSM). The discrete volume acquired represents a large mass of data and can be reduced with this particular method, before measurement (processing time) or archiving (memory space). Furthermore, the structure data is a powerful tool to rapidly compute parameters that are characteristic of the volume data. PMID- 8402525 TI - Visualization and interactive exploration of multidimensional confocal images. AB - A confocal image analysis system is developed for automatic extraction of surface representation of biological structures. A visualization system is also developed to manipulate these surface representations and to obtain morphometrical parameters and provides a powerful tool for biomedical research such as microstructural characterization, morphogenesis, cell differentiation, tissue organization, and embryo development. PMID- 8402526 TI - Method for 3D volumetric analysis of intranuclear fluorescence distribution in confocal microscopy. AB - The introduction of the confocal laser scanning microscope makes it possible to acquire fluorescent specimens in 3D. We present basic image processing tools to enhance the data and to quantitate the morphology and topography of the intravolume elements. The tools were applied to the description of the spatial distribution of DNA replication sites in mammalian cell nuclei as an example. PMID- 8402527 TI - The microanatomy of the alveolar duct of the human lung imaged by confocal microscopy and visualised with computer-based 3D reconstruction. AB - The most widely accepted model of human lung alveolar duct systems is that they are constructed from central helical fibres between the turns of which lie alveolar opening. Experimental difficulties of handling and sectioning lung tissue have made it difficult to confirm this. Confocal laser scanning microscopy (CLSM) was therefore used to generate optical serial sections of the lung that were reconstructed in three dimensions and displayed using volume rendering techniques. From images of the reconstructions, a new structure is proposed in which alveolar ducts consist of collections of connected oval, twisted loop structures with eccentric openings. PMID- 8402528 TI - Three-dimensional confocal microscopy and visualization of the in situ cornea. AB - The in situ cornea is an ideal test specimen to evaluate techniques for 3D reconstruction and visualization of unstained, unfixed, transparent living tissues from a stack of optical sections. The 0.4 mm thick transparent specimen has been optically sectioned into 365 sections using a confocal laser scanning microscope (CLSM) with a water immersion objective. Depth-dependent light attenuation due to absorption and scatter within the specimen was manually compensated at each sampled section. A water immersion microscope minimized the spherical aberrations that would have occurred with the use of an oil immersion objective. Isometric sampling resulted in near-cubic voxels, which compensated for the reduced microscopic resolution in the z axis as compared to x and y resolution. PMID- 8402529 TI - Dental anatomy portrayed with microscopic volume investigations. AB - The clinical treatment of the root canal of teeth--called endodontics--assumes a precise idea of the spatial arrangement of the anatomy of teeth and their inner structure. By using computer-assisted data acquisition from filmed sequences of histologic serial sections and a special kind of magnetic resonance microscope- the Stray Field Imaging (STRAFI)--volume investigations were carried out using special functions of a newly developed 3D software. Possible applications and future perspectives are discussed. PMID- 8402530 TI - Circulatory pattern and structure in the tail and tail fins of Xenopus laevis tadpoles. AB - This investigation was initiated with the intent of study capillary sprouting in the tadpole tail fin microcirculation of the African clawed frog, Xenopus laevis, using a combination of intravital video recordings and electron microscopy. The tadpoles were observed daily for periods up to one hour during one to two weeks. The capillary sprouts originated mostly from postcapillary venules, and within 24 48 h merged with other capillary sprouts, subsequently establishing a capillary loop with blood flow. As the circulatory patterns developed further, capillary regressions also occurred. As the electron microscope analyses of the capillary sprouts progressed, it became obvious that a thorough electron microscope study of the blood vessels of the tadpoles was required in order to explore structural characteristics of arterial and venous blood vessels. Thus, this article deals primarily with the general organization of the tadpole tail circulation, and the ultrastructural characteristics of the vascular walls. A subsequent article will deal with the role of endothelial cells, fibroblasts and pericytes in the process of capillary sprouting and regression, based on intravital recordings and electron microscope analyses. PMID- 8402531 TI - Ocular pathology in mice with a transgenic insertion at the microphthalmia locus. AB - A DNA insertional mutation at the microphthalmia locus (mi) in a transgenic mouse has been developed and given the allele symbol of mitg (Krakowsky et al., 1993). Mice homozygous for this transgene have eyes markedly reduced in size and relatively unpigmented. In this study, we examined the morphology of these eyes using light and electron microscopy. Transgenic homozygous (mitg/mitg) animals have a structurally normal choroid which lacks melanocytes but contains occasional leukocytes. The elastica of Bruch's membrane is absent except in an occasional site. The retinal pigmented epithelium (RPE) appears dramatically abnormal. It displays cellular heterogeneity, residual basal infoldings and apical microvilli, rare and immature melanosomes, and numerous cilia-like structures. Occasionally, cells of the RPE appear to have extruded into or from the choroid. The photoreceptor cells are devoid completely of outer segments and partially of inner segments. Numerous active macrophages are present between the amelanotic RPE and neuro-retina and also within the vitreous body. The anterior uveal tract is underdeveloped and hypomelanotic. This new microphthalmia model exhibits ocular pathology with similarities and differences to other mutations and the mi (microphthalmia) locus. PMID- 8402532 TI - Diversity of cell-cell interactions formed by gastric parietal cells in culture: morphological study on guinea pig cells. AB - Parietal cells of gastric glands are specialized to produce acid. Tight junctions between the parietal cells and their neighbouring cells (usually chief cells and mucous cells, less commonly parietal cells) avoid acid back-diffusion. Alterations of these junctions are accompanied by a defective epithelial barrier function. The conditions leading to junction formation, e.g. during epithelial restitution and regeneration are entirely unknown. The present study has the purpose to establish an in vitro model which allows studying these junctions. Freshly isolated gastric epithelial cells of guinea pig, moderately enriched with parietal cells, were cocultured for 2 days. Highly specific staining techniques showed the following composition in the near-confluent monolayer: 45% parietal cells (succinic dehydrogenase-positive), 36% mucous cells (lectin-binding granules), 18% chief cells (pepsinogen-positive granules) and 1% subepithelial cells (vimentin-positive). Ultrastructural investigations of sections of these monolayers revealed a high tendency of parietal cells to form cell junctions with the following characteristics: 1) virtually all parietal cells formed junctions with their neighbouring cells; 2) only junctions, but no desmosomes, were observed among neighbouring parietal cells; 3) junctional complexes and desmosomes were regularly present between parietal cells and their neighbouring mucous and chief cells; 4) parietal cells were sometimes integrated into three dimensional structures, resembling rudimentary gastric glands. In conclusion, parietal cells under standard coculture conditions, generate de novo the same types of cell junctions that are observed in the intact gastric epithelium. The results show that parietal cells in vitro spontaneously make junctions with parietal and non-parietal cells, resembling the junctions in the intact tissue. PMID- 8402533 TI - In situ hybridization at the ultrastructural level: localization of cytomegalovirus DNA using digoxigenin labelled probes. AB - We describe a procedure for detecting and localizing cytomegalovirus DNA sequences based on in situ hybridization at the ultrastructural level. A digoxigenin-labelled probe, identified with an anti-digoxigenin colloidal gold labelled antiserum, was employed on infected cells embedded in a new acrylic resin (Bioacryl). The silver enhancement method on the same specimen was used to more easily reveal the reaction also at low magnification. The immunolocalization was characterized by high specificity with virtually no background staining and a good maintenance of submicroscopic cell features. PMID- 8402534 TI - Muramidase-mediated damage to Candida yeast cells. Histochemical and immunochemical characterization of accumulating wall-like material. AB - Hen egg-white lysozyme is known to be fungicidal to blastoconidia of Candida albicans under defined in vitro conditions. This lethal action leads to changes in the layering of cell wall and to plasmolysis, caused by unremitting accumulation of wall-like material between the yeast cell wall and cytoplasmic membrane. Here, several methods were applied on ultrathin sections to define the nature of wall-like material: histochemical staining with periodic acid thiocarbohydrazide-silver proteinate, periodic acid-alkaline bismuth, and phosphotungstic acid at low pH; the localization of the carbohydrate residues with lectin-gold complex; immunocytochemical staining with monospecific antibodies, factor 1 and 6, which recognized major cell wall antigens. The wall like material was almost uniformly highlighted with periodic acid thiocarbohydrazide-silver proteinate, factor 1 antibody, concanavalin A-gold and wheat germ agglutinin-ovomucoid-gold, indicating the presence of mannoproteins and chitin. The serotype A-specific epitope recognized by factor 6 antibody was not detected in the wall-like material, although it was demonstrated in the outer cell wall layers after 2 h of exposure to lysozyme. PMID- 8402535 TI - Morphological changes of myenteric neurons in the partially obstructed opossum esophagus. AB - We have previously described a model of partial esophageal obstruction which leads to hypertrophic and degenerative changes in the smooth muscle cells. In the present communication we describe changes occurring in the myenteric plexus including increased thickness, varicosities and irregular angulations of nerve fiber bundles. The perikarya of myenteric neurons in the obstructed esophagus showed elongated mitochondria, dilated cisternae of the endoplasmic reticulum and increased numbers of secondary lysosomes and multimembranous dense bodies. In myenteric axons, there was infiltration of Schwann cell subunit by collagen fibrils. PMID- 8402536 TI - The effect of insulin on the liver cells of newborn rats. An electron microscopic study. AB - The liver of newborn rats contains neither glycogen nor lipid droplets. These latter, sometimes in fusion, still could be found in the cytoplasm and the nucleus as well, following a five-minute action of insulin at a dose of 0.2 IU/animal. The lipid droplets were in close relationship with the mitochondria. This time the glycogen, either in fields or rosettes, was missing. Thirty minutes after treatment the fields of glycogen could be well seen. The experiments demonstrated an exclusive sequence of events starting with the receptor and its signal insulin on neonatal liver cells and resulting in a morphological picture (involving first lipid droplets, then glycogen), similar to that of the adult liver cells. PMID- 8402537 TI - Ultrastructural sperm abnormalities and cerebellar atrophy: does a correlation exist? Report of two cases without endocrine hypogonadism. AB - Spermiograms were performed in two young patients with cerebellar ataxia, one familial and the other sporadic. The subjects did not have endocrine abnormalities, but there was MRI evidence of cerebellar atrophy. Light and electron microscope examination revealed sperm abnormalities similar to those described in Purkinje cell degeneration (PCD). PCD is an autosomal recessive mutation observed only in the mouse, characterized by mild ataxia due to the postnatal degeneration of cerebellar Purkinje cells and male sterility due to morphological sperm abnormalities. PMID- 8402538 TI - Immunophenotype and ultrastructure of alveolar soft part sarcoma. AB - Nine cases of alveolar soft part sarcoma were studied in terms of their immunoreactivity towards intermediate filaments and muscle markers, and their ultrastructure with regard to muscle features and the cytoplasmic crystals characteristic of this tumour. Inconsistent expression of alpha-smooth muscle actin, vimentin and desmin were found. No unambiguous muscle cell characteristics were identified by electron microscopy, although multiple subplasmalemmal densities with overlying lamina were emphasised as a typical but not specific muscle feature. Classical alveolar soft part sarcoma crystals at the ultrastructural level were absent throughout, but pseudo-crystals with a pre crystalline finely filamentous content were numerous in one case. The results were integrated into a comprehensive comparison with the literature. It is clear that immunohistochemical evidence of striated muscle differentiation is detectable in some but not all cases of ASPS. The detection of Myo D1 protein in the small numbers of ASPSs so far studied for this gene product raises the possibility that in the majority of ASPSs this is the only evidence of early striated muscle differentiation and that only in the minority is it able to induce desmin production. Study of a substantial series for Myo D1 is required to explore this possibility. PMID- 8402539 TI - A novel type of microtubules in the frog urinary bladder epithelium stimulated by vasopressin. AB - Changes in the cytoskeleton of granular cells of the frog urinary bladder under conditions of low and vasopressin-induced water permeability were investigated. Using a conventional electron microscopical preparation with glutaraldehyde/osmium fixation we found only the usual type of microtubules in granular cells of both resting and stimulated bladders. These microtubules, 20 nm in diameter, were associated with centrioles and Golgi complexes. Under the high water permeability conditions the large vacuoles appear in granular cells. The 20 nm microtubules and microfilaments were connected to the membrane of these vacuoles. The second type of 'thick' microtubules was found in granular cells of the stimulated bladder after the fixation with glutaraldehyde without osmium postfixation or freeze-substitution. The tubulin nature of these 'thick' microtubules was identified immunocytochemically. In addition, tubulin was present in the granules with crystalline content. PMID- 8402540 TI - The morphology of teleost meningocytes as revealed by freeze-fracture. AB - We describe the morphology of the cells in the three layers of the teleost endomeninx, as viewed by freeze-fracturing. The cells of the outer endomeningeal layer are fusiform and closely packed, with interdigitations that hold the cells together, gap junctions and a few strands of particles resembling tight junctions, but no desmosomes. The intermediate layer is formed by a single layer of flattened and elongated cells with rectangular shape, and well developed junctional complexes (gap junctions, tight junctions and desmosomes). These cells also show numerous plasmalemmal vesicles (6.5 +/- 1.3/microns 2) in the upper (in contact with the outer layer) and lower (in contact with the inner layer) membranes. Cross fracture of these cells shows many membrane-bound and free vesicles. The inner layer is formed by spindle shaped cells with wide intercellular spaces filled with a granular matrix and collagen fibers. The density of intramembrane particles is higher than that in the meningocytes of the outer and intermediate layers. The morphology of the teleost endomeninx appears similar to that of the leptomeninges of birds and mammals, but the outer endomeningeal layer of teleosts (which resembles the arachnoid of elasmobranchs and amphibians) appears different from any cell layer in the meninges of amniotic vertebrates. PMID- 8402541 TI - Presence of a crystal in the cytoplasm of the male germ cells of the garden dormouse Eliomys quercinus L. AB - A crystal is present in the cytoplasm of the male germ cells of a hibernator, the garden dormouse, Eliomys quercinus. In another hibernator, the thirteen-lined ground squirrel Citellus tridecemlineatus, such crystals were not found. In the garden dormouse, the crystal is present in the cytoplasm of spermatocytes I and II and of spermatids, after which it is found in the residual bodies. The crystal is usually an ovoid ca 2.4 microns long and 0.75 microns by 1.6 microns when sectioned transversally at its widest region. It is composed of 40 to 80 tubule like structures, which are 12 nm in outside diameter and have a lumen 8 nm in diameter; the wall is thus ca 2 nm thick. Each tubule seems to be made of two fibrils ca 2 nm in diameter tightly coiled. The tubule-like structures have a zigzag course and are parallel to one another, about 8 nm apart; they are arranged like stacked spoons. Each tubule-like structure consists of two repeating units, one 40 nm long and the other 60 nm. Each unit is inclined to its predecessor at 130 degrees, alternately right and left, thereby giving the zigzag effect. The crystals are present in active and in torpid animals and during normal or induced spermatogenesis; their presence, therefore, does not seem to be dependent on the hormonal status of the hibernator. PMID- 8402542 TI - The fine structure of Gastromermis sp. (Nematoda: Mermithidae) sperm. AB - The development and morphology of the sperm of the mermithid nematode, Gastromermis sp. appears to be different than that reported for any other nematode sperm. The mature sperm of Gastromermis are elongate with membranous organelles surrounding an apical nuclear region. The mitochondria are arranged in parallel rows and form an elongate sheath. Spermatozoa contain dense tubular structures which have not been seen in other nematodes. Spermatids lack fibrous bodies. The pseudopodia of the spermatozoa are filled with microtubules. Centrioles seen throughout sperm development are composed of 9 doublets. PMID- 8402543 TI - Lipid-rich residual bodies and cathepsin D in the human uterus: ultrastructural and quantitative comparison between normal myometrium and leiomyoma. AB - The lipid-rich residual bodies (LRRB) (Eyden et al., 1991) in human myometrium and uterine leiomyoma cells, have a distinctive ultrastructure characterised by a rich lipid content. To evaluate the biological or pathological significance in detail, normal myometrium and uterine leiomyoma from 30 human cases were studied by conventional histological, histochemical, immunohistochemical and electron microscopic methods. The study included a quantitative analysis of LRRBs of 3 premenarchic cases, 19 cases having a menstrual cycle, and 8 cases in menopause, in addition to 20 patients with histologically conventional leiomyoma larger than 3 cm in diameter. The study revealed the following findings: 1) immunohistochemical distribution of cathepsin D in the LRRB; 2) histochemical demonstration of neutral fat as the main content of LRRB; 3) statistically significant decrease in the distribution of LRRB in leiomyoma tissue compared with normal myometrium; 4) an absence or minimal distribution of LRRB in premenarchic myometrium; 5) a moderately significant correlation between the frequency of LRRB and patient's age. The distribution of cathepsin D within LRRB and the differential expression of LRRBs in the various smooth muscle cell tissues of the uterus suggest a possible role of ovarian hormones in the genesis of LRRBs which may function in the intra-lysosomal degradation of organelles produced during hormonal cycling. PMID- 8402544 TI - Role of the gene on trisomic and pentasomic chromosome 13 in murine mammary tumorigenesis. AB - To study a possible role(s) played by the trisomy and pentasomy of chromosome 13 in murine mammary tumors, we examined, in eight cloned established cell lines derived from a single BALB/c mammary tumor induced by MTV, a correlation between the presence of trisomy or pentasomy 13 and transformation parameters and in vivo tumorigenicity in syngeneic mice. We found that cell lines with a higher incidence of trisomy or pentasomy 13 in cells of diploid and tetraploid chromosome numbers, respectively, grew to a much higher cell density in flasks than did those with low incidence, and they formed tumors in syngeneic BALB/c mice, whereas those with a low incidence of trisomy or pentasomy 13 were poorly tumorigenic. The presence in the tumorigenic cells of trisomy or pentasomy 13 was not correlated with their growth in soft agar. Furthermore, other chromosomes manifested a wide range of copy numbers in the presence of trisomy or pentasomy 13, indicating that no chromosomes counteracted chromosome 13 to prevent the tumorigenicity. In light of the tumorigenic growth of the cells that maintain gene dosage of chromosome 13 at different ploidy levels, the possibility of the yeast G1 cyclin-like roles played by the gene(s) residing on chromosome 13 in murine mammary tumorigenesis is discussed. PMID- 8402545 TI - Cytogenetic homogeneity in eight independent sites in a case of malignant melanoma. AB - Melanoma cell lines initiated from metastases excised at the same time from multiple sites in a patient reflect a single clone origin. The similarities of the karyotypes of these lines depend upon the site of excision, the selective survival in vitro, and the time span in vitro before study. In the present investigation, eight short-term cultures of malignant melanoma from the same patient were analyzed cytogenetically, six after the second passage in vitro, one from the third, and one from the fifth passage. Karyotypes were similar on all eight cultures, each having chromosome abnormalities der(1)t(1q;?), der(3)t(1;3), and t(7;9). Four cell lines had an isochromosome 4q, which was missing in four lines. Two cell lines were missing del(6). In five of eight cultures there was an occasional i(16q) or t(16q;21q). In the final culture, made 6 months after the first seven, three cell types present included a type corresponding to that of seven previous cultures, a similar cell type with an abnormal chromosome 11, and a third cell type with a 3;19 translocation. Comparisons are made with similar studies in the literature and a hypothesis has been formulated to explain melanoma metastases as revealed by the cytogenetics of multiple lesions excised simultaneously. PMID- 8402546 TI - Detection of the PML/RAR alpha fusion gene in acute promyelocytic leukemia with a complex translocation involving chromosomes 15, 17, and 18. AB - We report a case with typical clinical features of acute promyelocytic leukemia (APL) carrying an atypical chromosomal aberration involving chromosomes 15, 17, and 18. Molecular analysis using Southern blot hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR) proved the creation of the PML/RAR alpha fusion gene in this case. These findings support the notion that this fusion is of crucial importance to leukemogenesis of APL. PMID- 8402547 TI - Simple numerical chromosome aberrations in two pituitary adenomas. AB - Cytogenetic analysis of short-term cultures of one non-secreting and one prolactin-producing pituitary adenoma revealed simple clonal numerical abnormalities in both tumors. The karyotype of the non-secreting adenoma was 48,XX, +4, +9[42]/49,XX, +4, +9, +20[2]/46,XX[6]. In the prolactin-secreting adenoma, three aberrant clones were detected, giving the karyotype 45,X, Y[20]/47,XY, +Y[6]/45,XY, -21[3]/46,XY[21]. One cell had the chromosome complement 46,X, -Y, +9; no other nonclonal aberrations were detected. The only hitherto published case of pituitary adenoma analyzed by banding techniques (Rey et al. [1986]: Cancer Genet Cytogenet 23:171-174) also had only numerical clonal changes that included extra copies of chromosome 9. We conclude that pituitary adenomas may be karyotypically characterized by numerical aberrations and that trisomy 9 seems to be the best candidate for a primary chromosomal anomaly. PMID- 8402548 TI - Mapping of the chromosome 11 breakpoint of the t(4;11)(q21;p14-15) translocation. AB - We describe the localization of the chromosome 11 breakpoint of a T-ALL translocation, t(4;11)(q21;p14-15), to sub-band 11p15.5. The breakpoint is located between the genes for insulin-like growth factor II (IGFII) and the M1 subunit of ribonucleotide reductase (RRM1). This region does not include any previously cloned genes involved in cancer. PMID- 8402549 TI - A further case of acute nonlymphocytic leukemia with tetrasomy 8. AB - In very rare patients tetrasomy of chromosome 8 is observed as the only karyotypic event. We present a 74-year-old man with acute monocytic leukemia (FAB M5) displaying 46,XY/48,XY, +8, +8 in unstimulated bone marrow cells. The patient died 2 weeks after diagnosis. Tetrasomy 8 appears to identify a subgroup of acute nonlymphocytic leukemia. It may indicate a poor prognosis. PMID- 8402550 TI - Evolution of complex chromosomal rearrangements in a case of biphenotypic pre B/myeloid acute leukemia. AB - A case of acute biphenotypic leukemia, in which blast cells coexpress antigens of both lymphoid and myeloid lineages, was seen in a 24-year-old female. The clonal karyotype was complex but did not include any of the abnormalities previously found in biphenotypic leukemia. Chromosome abnormalities included breakpoints seen in both acute lymphoblastic and acute myeloid leukemias as well as a number of known fragile sites, namely the rare heritable fragile site at 10q24, and the common, aphidicolin-induced sites on chromosome 7. PMID- 8402551 TI - Involvement of 10q22 in leiomyoma. AB - Cytogenetic investigation of uterine leiomyomas revealed a rearranged 10q22 present in nine tumors as a clonal change in most of the cells analyzed. These findings indicate that abnormalities involving chromosome 10 and, particularly 10q22, may characterize a cytogenetic subgroup of uterine leiomyomas. PMID- 8402552 TI - Trisomy 13 in a patient with leukemic progression of myelodysplasia. AB - Four months after the diagnosis of refractory anemia, a 60-year-old patient developed acute leukemia with blast cells that were poorly differentiated by morphology and clearly myeloid by immunophenotyping. Cytogenetic analysis performed at leukemization showed trisomy 13. An extra copy of chromosome 13 has already been reported in a few cases of acute leukemia and myelodysplastic syndrome. PMID- 8402553 TI - Trisomy 4 may occur in a broad range of hematologic malignancies. AB - Trisomy 4 is an uncommon numerical chromosomal aberration in acute leukemia. We describe three cases of trisomy 4, occurring in two patients with acute myeloid leukemia (M1 and M5a) and in one patient with T-cell acute lymphoblastic leukemia. Our results suggest that trisomy 4 may occur in a broader range of hematologic and malignancies than previously described. PMID- 8402554 TI - Telomeric fusions in a Wilms' tumor. AB - The present report describes the karyotypic findings in cells from a Wilms' tumor. The most consistent cytogenetic abnormalities detected consisted of translocations involving break and fusion of chromosomal telomeres and telomeric associations frequently affecting the terminus of the short arms of chromosomes 14 and 17. PMID- 8402555 TI - Chromosomes in the genesis and progression of ependymomas. AB - Cytogenetic analysis was performed on cultures of primary ependymal tumors with different degrees of malignancy (I-IV) obtained from four patients, none of whom had received therapy before karyotypic evaluation. The most common abnormalities were monosomy 17 and 22 in four cases and losses of sex chromosomes in three cases. Structural rearrangements of chromosome 2 were a finding for all cases and involved loss of material at 2q32-34. Other structural chromosome abnormalities detected involved chromosomes 4, 6, 10, 11, 12, and X. We also reviewed data on 22 cases previously reported. PMID- 8402556 TI - A complex translocation, t(4;11;13)(q21;q23;q12-14), in a case of infantile acute lymphoblastic leukemia. AB - Cytogenetic analysis provides valuable prognostic information in children diagnosed with hematologic malignancies. While the t(4;11)(q21;q23) has frequently been reported in patients with acute lymphoblastic leukemia, the additional involvement of chromosome 13 to form a three-way translocation has not been described previously. We report a case of acute lymphoblastic leukemia in a 5-month old infant with a complex t(4;11;13)(q21;q23;q12-14). PMID- 8402557 TI - Acute myelomonocytic leukemia in a XYY man. AB - A case of acute myeloid leukemia (M4) in a 29-year-old male with a 47,XYY karyotype is reported. This aneuploidy was found in both bone marrow cells and mitogen-stimulated lymphocytes. Monosomy 7 correlated with myelodysplastic features. The possible role of XYY in increasing the risk of leukemia is discussed. PMID- 8402558 TI - A novel translocation (3;11)(q26;q13) in a case of acute myelomonocytic leukemia. AB - We report a novel translocation (3;11)(q26;q13) in a case of acute myelomonocytic leukemia. The implications of rearrangements involving chromosomes 3q26 and 11q13 are discussed. PMID- 8402559 TI - Chromosome 12 alterations and c-Ki-ras mutations in colorectal tumors. AB - c-Ki-ras mutations and gains of chromosome 12, where this gene is mapped, were both studied in a series of 47 colorectal cancers. Mutations at codon 12 and 13 were detected in 17 (36%) and gains of chromosome 12 in 7 (15%) cases. In this sample, gains of chromosome 12 occur in tumors either with or without c-Ki-ras mutations, suggesting that gains of chromosome 12 are independent from the mutation of c-Ki-ras. PMID- 8402560 TI - Association between t(2;9)(p12;p23) and early B-precursor acute lymphoblastic leukemia. AB - We report the occurrence of t(2;9)(p12;p23) in a 20-month-old girl with early B precursor acute lymphoblastic leukemia (ALL). This translocation has only been reported once before in an adult case of early B-precursor ALL with t(4;11)(q21;q23). We suggest that t(2;9)(p12;p23) may be associated with this particular phenotype of ALL. PMID- 8402561 TI - A case of monosomy 20 in an adult acute lymphoblastic leukemia. PMID- 8402562 TI - High incidence of constitutional balanced translocations in neuroblastoma. PMID- 8402563 TI - Biologic and clinical significance of cytogenetic and molecular cytogenetic abnormalities in benign and malignant cartilaginous lesions. AB - Cartilaginous neoplasms are often histologically and therapeutically challenging. Predicting biologic behavior can be difficult. In this study, 120 nonneoplastic, benign, and malignant cartilaginous lesions from 103 patients were cytogenetically analyzed in a 6-year period after short-term culture. For selected cases, fluorescent in situ hybridization (FISH) techniques using chromosome-specific probes were performed on metaphase/interphase preparations and on paraffin-embedded tissue sections. Clonal abnormalities of chromosomes 2, 3, 5, 7, 8, and 12 were most frequently observed. Involvement of chromosomes 5, 8, and 12 may be etiologically significant because of the gene localizations for the human cartilage link protein, Langer-Giedion syndrome (a rare syndrome characterized by multiple exostoses), and type II collagen (a major component of normal cartilage) respectively, to these three chromosomes. That chromosome 7 abnormalities were observed only in malignant tumors is of diagnostic value. The identity of three marker chromosomes and the significance of trisomy 7 (a finding of controversial meaning), were determined with FISH. That the presence of chromosome aberrations and increasing histologic grade strongly correlated (p = 0.001) is of prognostic importance. Moreover, complex aberrations were observed nearly exclusively in high-grade tumors (p = 0.001). The data show that nonrandom chromosome loci are aberrantly affected in cartilaginous lesions and that these abnormalities may be of significant histopathogenetic consequence. In addition, these chromosome abnormalities appear to be diagnostically and prognostically valuable in classifying and grading chondromatous neoplasms. PMID- 8402564 TI - Direct chromosome analysis of 50 primary breast carcinomas. AB - Direct chromosome analyses were performed in 50 cases of primary breast carcinoma. Thirty-six cases had modal chromosome counts in the diploid range; the other 14 cases were polyploid. Of the 22 cases with detailed G-banding analyses, the most frequent structural changes involved chromosome 1 (15 of 22), 3 (13 of 22), and 6 (13 of 22). Deletion of chromosome 1p was noted in nine cases, and both 3p- and 6q- were noted in 10 cases. PMID- 8402565 TI - Metabolism of [1-14C]linoleic acid in human promyelocytic leukemia HL-60 cells grown and differentiated in serum-free medium. AB - The metabolism of [1-14C]linoleic acid (LA) was studied in human promyelocytic leukemia cell line HL-60, grown and differentiated in serum-free medium. Both undifferentiated and dibutyryl cyclic AMP-differentiated HL-60 cells exhibited similar patterns of conversion of LA to four other major fatty acids (i.e. 18:3, 20:3, 20:4, and 22:4). PMID- 8402566 TI - Dose-related inhibition by dietary phenethyl isothiocyanate of esophageal tumorigenesis and DNA methylation induced by N-nitrosomethylbenzylamine in rats. AB - The purpose of this investigation was to establish a dose response for the effects of dietary phenethyl isothiocyanate (PEITC) on N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumorigenesis and DNA methylation. Groups of 13-27 rats were randomly assigned to AIN-76A diets containing 0, 0.325, 0.75, 1.5 or 3.0 mumol PEITC/g. Two weeks later, rats were administered NMBA subcutaneously at a dose of 0.5 mg/kg once a week for 15 weeks. Animals were maintained on control or experimental diets for an additional 8 weeks and were terminated at week 25 of the experiment. No significant effects on weight gain or food intake were noted for any of the experimental diets when compared with control values. Animals receiving only NMBA developed 9.3 +/- 0.9 tumors/rat, with an incidence of 100%. Dietary PEITC at concentrations of 0.75, 1.5 and 3.0 mumol/g inhibited NMBA induced esophageal tumor multiplicity by 39%, 90% and 100%, respectively. Esophageal tumor incidence in these groups was reduced by 0%, 40% and 100%, respectively. The 0.325 mumol/g PEITC diet did not significantly affect NMBA induced esophageal tumorigenesis. These results indicate that the minimum inhibitory dietary concentration of PEITC is between 0.325 and 0.75 mumol/g. Groups of 20 rats were assigned to diets containing 0-3.0 mumol PEITC/g for two weeks as described above, and then sacrificed 24 hours after administration of [3H-methyl]NMBA. The esophageal DNA was isolated, purified, hydrolyzed, and analyzed by HPLC. PEITC inhibited DNA methylation in a dose-dependent manner, as was found in the tumor bioassay. The inhibition of tumor incidence was highly correlated with the percentage inhibition of either 7-methylguanine or O6 methylguanine. These latter results suggest that the inhibitory activity of PEITC in this model is manifested, at least in part, during the functional equivalent of tumor initiation. PMID- 8402567 TI - Influence of ascorbic acid on MCA-induced carcinogenesis in the uterine cervix of mice. AB - The present study evaluates the possible modulatory influence of ascorbic acid (AA) on (methylcholanthrene) MCA-induced cervical carcinogenesis in mice. Ascorbic acid was given daily for different durations in drinking water at the dose level of 2 mg/ml. In animals that received no modulator, placement of cotton threads impregnated with beeswax containing -600 micrograms of MCA for 16 weeks yielded 76.1% carcinomas in the uterine cervix. Administration of AA for the entire period of 16 weeks resulted in the reduction of MCA-induced cervical carcinomas to 33.3%. AA was capable of reducing cervical cancer incidence to 33.3% even when it was given for only 6 weeks following MCA-thread insertion into the uterine cervix. However, when it was given for a period of 10 weeks after 6 weeks of MCA-thread insertion the carcinoma incidence was as high as 66.6%. Results of the present study demonstrate that AA, especially when given during the initiational phase, significantly inhibits MCA-induced cervical carcinogenesis in mice. PMID- 8402568 TI - Immunoperoxidase localization and denovo biosynthesis of a 10.5-kDa inhibin in benign and malignant conditions of human breast. AB - Using immunocytochemistry, we report the occurrence of a 10.5-kDa inhibin in human breast tissue specimens obtained from normal, fibroadenoma and adenocarcinoma cases. The immunostaining for inhibin was confined to the cytoplasm of the epithelium and myoepithelium cells. Expression of inhibin increased in following order: normal (1+); adenocarcinoma and lobular carcinoma in situ (2+) and fibroadenoma (4+). Breast explants has the ability of denovo biosynthesis of inhibin in vitro. In view of the growth modulating regulatory properties of 10.5 kDa inhibin, our findings are suggestive of the potential role of inhibin in breast pathology. PMID- 8402569 TI - Cancer incidence differences between ethnic groups in Surinam. AB - Because of central registration, all new cases of malignant disease in Surinam confirmed by pathology could be studied over a twelve-year period (1979-1990). Patients were arranged by sex, age, racial or ethnic group, and primary localisation of the malignancy. A predominance of female patients could largely be ascribed to very frequent carcinoma of the uterine cervix. Apart from this, striking differences in proportional rate of general malignancy and of the four most frequent types were found among the various ethnic groups. Genetic differences in susceptibility to malignant disease are presumed to contribute substantially to the variations observed. PMID- 8402570 TI - Effect of glucocorticoids on TPA-induced inhibition of gap-junctional communication and morphological transformation in Syrian hamster embryo cells. AB - The effect of glucocorticoids on 12-O-tetradecanoylphorbol-13-acetate (TPA) induced inhibition of gap-junctional intercellular communication (GJIC) and morphological transformation in Syrian hamster embryo (SHE) cells was examined. Fluocinolone acetonide (FA) and dexamethasone (DEX) almost completely suppressed the effect of TPA on induction of transformed morphology. On the other hand, up to 1000 times higher FA and DEX concentrations did not influence the inhibitory effect of TPA on GJIC. Neither treatment with these glucocorticoids for 4, 24 or 48 h before TPA exposure nor 24 h co-exposure with TPA altered the effect of TPA on GJIC. Thus the potent effect of glucocorticoids as inhibitors of the promotional effect of TPA on morphological transformation in SHE cells does not result in alterations of TPA-induced inhibition of GJIC. PMID- 8402571 TI - Characterization of phospholipase A2 activity in MDA-MB-435 human breast cancer cells. AB - Phospholipase A2 (PLA2) was identified and its properties characterized in MDA-MB 435 cells, a human breast cancer cell line. Cytosolic fractions, prepared in calcium-free buffer, were assayed using arachidonyl-containing phosphatidylcholine as substrate. PLA2 activity was linear as a function of both time and protein concentration. The enzyme was shown to be calcium-dependent and to require a basic pH of 9.5-10.0 for optimal activity. Activity was predominantly found in the cytosolic fraction when cells were harvested in calcium-free buffer. Phospholipase A2 may play a key role in linoleic acid enhanced mammary tumorigenesis and metastasis. PMID- 8402572 TI - Sensitivity of human colon tumor metastases to anticancer drugs in athymic (nude) mice. AB - We have studied the metastatic behavior of five human colon tumor cell lines (LS174T, WiDr, Caco-2, SW 620 and SW 480) using intrasplenic-nude mouse (ISMS) and intravenous-nude mouse (IVMS) model systems. LS174T was highly metastatic in both systems. In the IVMS system, LS174T cells produced lung metastasis and also grew in the skin as if the cells had been injected subcutaneously. We have also studied the anti-metastatic activity of three anticancer drugs (5-fluorouracil (5 FU), doxorubicin (DX) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) using LS174T cells and both ISMS and IVMS systems. The drugs were given intravenously on day 19 and 26 of tumor cell injection. Mice were sacrificed and organs observed for metastatic growth 4-6 weeks after cell injection. The results show that in the ISMS system, BCNU and 5-FU are inactive against both the liver metastasis and primary growth in the spleen. DX inhibits metastatic growth but not the primary growth. In the IVMS system, BCNU is inactive, whereas 5-FU and DX are active against the metastatic growth. Thus, DX may have activity against blood-borne human colon tumor metastasis. PMID- 8402573 TI - Effect of arecanut on the black mustard (Brassica niger, L.)-modulated detoxication enzymes and sulfhydryl content in the liver of mice. AB - The modulatory potential of arecanut, a popular masticatory, was assessed on the black mustard-induced changes in hepatic detoxication system in mice. The modulatory effect was assessed on glutathione S-transferase (GST), cytochrome b5 (Cyt. b5) and cytochrome P-450 (Cyt. P-450) and acid-soluble sulfhydryl (-SH) content. Mice were fed on either normal diet or diet containing 0.25%, 0.5% or 1% (w/w) arecanut for 45 days. During the last 10 days of treatment the feed was supplemented with 0.5% or 1% black mustard, the popular condiment. Dietary feeding of mustard could significantly enhance the studied phase I and phase II enzymes as well as -SH content in murine liver. However, black mustard-induced alterations in GST and -SH content were lower, while Cyt. b5 and Cyt. P-450 levels were much higher in mice receiving arecanut treatment than controls. PMID- 8402574 TI - Evaluation of the modulatory influence of black pepper (Piper nigrum, L.) on the hepatic detoxication system. AB - The present paper assesses the modifying potential of black pepper on the hepatic biotransformation system in mice. The modulatory effect was assessed on glutathione S-transferase (GST), cytochrome b5 (cyt. b5), cytochrome P-450 (cyt. P-450), acid-soluble sulfhydryl (-SH) content and malondialdehyde (MDA) level. Swiss albino mice of either sex (eight weeks old) were fed on a diet containing 0.5%, 1% and 2% black pepper (w/w) for 10 and 20 days. The findings revealed a significant and dose-dependent increase in GST and -SH content in the experimental groups except the one maintained on 0.5% black pepper diet for 10 days. Elevated levels of cyt. b5 and cyt. P-450 were also statistically significant and dose-dependent. The level of MDA was lowered in the group fed on 2% black pepper diet for 20 days. Being a potential inducer of detoxication system, the possible chemopreventive role of black pepper in chemical carcinogenesis is suggested. PMID- 8402575 TI - Effect of modulation of protein kinase C activity on cisplatin cytotoxicity in cisplatin-resistant and cisplatin-sensitive human osteosarcoma cells. AB - The effect of modulation of protein kinase C (PKC) activity by 12-O tetradecanoylphorbol-13-acetate (TPA) on cisplatin cytotoxicity was examined in a human osteosarcoma U2-OS cell line and in a U2-OS variant (U2-OS/Pt) selected after continuous exposure to increasing concentrations of cisplatin. U2-OS/Pt cells showed a 7.5-fold resistance to the drug. A 24 h exposure of cells to TPA caused a potentiation of cisplatin cytotoxicity in sensitive and in resistant cells; under these conditions, PKC activity was shown to be down-regulated. In contrast, a short-term exposure of cells to TPA did not affect cisplatin cytotoxicity in U2-OS or in U2-OS/Pt cells. These results support the involvement of PKC in cellular response to cisplatin. However, this enzyme is probably not directly implicated in the mechanisms of acquired resistance in this cell system. PMID- 8402576 TI - Enhancement by non-mutagenic pesticides of GST-P positive hepatic foci development initiated with diethylnitrosamine in the rat. AB - The potential hepatocarcinogenicity of seven pesticides was examined using a rapid bioassay based on the induction of glutathione S-transferase placental form positive foci in the rat liver. Rats were initially injected with diethylnitrosamine and two weeks later were fed on diet supplemented with one of the pesticides for 6 weeks and then killed; all rats were subjected to a partial hepatectomy at week 3. Positive results were seen with chlorobenzilate (2000 ppm), vinclozolin (2000 ppm), malathion (4000 ppm), tecnazene (2000 ppm) and isoproturon (2000 ppm). S,S,S-tributylphosphorotrithioate (DEF, 200 ppm) and dicloran (2000 ppm) were negative in both number and area analyses. Although chlorobenzilate is carcinogenic in mice, malathion and vinclozolin have been reported as non-carcinogens in both rats and mice. Since the present system is based on the two-stage carcinogenesis hypothesis, it is possible that the chemicals showing positive results in this system possess at least tumor promoting activity in the rat liver. This is very significant, as most carcinogens show tumor-promoting activity in their target organs. PMID- 8402577 TI - Detection of serum cytokine levels in experimental cancer cachexia of colon 26 adenocarcinoma-bearing mice. AB - The aim of this study was to evaluate the correlations between tumor size and cachexia parameters including cytokine levels in serum. In transplantable colon 26 adenocarcinoma-bearing mice, parameters having negative correlations with tumor size were host weight changes, epididymal adipose tissue weight, glucose and interleukin 3 (IL-3) concentration in serum. Parameters having a positive correlation with tumor size were the number of circulating white blood cells and immunosuppressive acidic protein (IAP), interleukin 6 (IL-6) and transforming growth factor beta (TGF-beta) concentration in serum. PMID- 8402578 TI - Tumour necrosis factor-alpha alters the blood compartmentation of amino acids in the rat. AB - The work presented focuses on the importance of studying the distribution of blood amino acids in both the cellular and plasmatic fraction when performing studies concerning the effects of cytokines on amino acid metabolism. Tumour necrosis factor treatment resulted in important changes in blood amino acid compartmentation between plasma and red blood cells in rats. The animals showed a change in compartmentation with an increase in the concentration of most amino acids in the cellular fraction with the exception of phenylalanine, glutamate, aspartate and tyrosine. PMID- 8402579 TI - Inhibition of human brain tumor cell growth by a receptor-operated Ca2+ channel blocker. AB - SK&F 96365, a reported receptor-operated Ca2+ channel blocker, inhibited the growth of U-373 MG human astrocytoma and SK-N-MC human neuroblastoma cell lines in a dose-dependent manner. Carbachol and serum which act as growth factors for these cells induced a rapid, transient increase of intracellular Ca2+ concentration without a sustained increase. SK&F 96365 also exerted a significant inhibition of carbachol or serum-induced intracellular Ca2+ mobilization. These results suggest that SK&F 96365 is a potent inhibitor of brain tumor cell growth and that its effect may be mediated by the inhibition of agonist-induced intracellular Ca2+ mobilization. PMID- 8402580 TI - Alterations in repair of alkylating agent-induced DNA damage in polyamine depleted human cells. AB - Treatment of HeLa cells with the polyamine biosynthesis inhibitors difluoromethylornithine (DFMO) and/or methylglyoxal bis(guanylhydrazone) (MGBG) results in marked depression in levels of the cellular polyamines putrescine, spermidine and spermine. Cells in this polyamine-depleted state exhibited increased sensitivity to monofunctional alkylating agents, manifested as decreased cloning ability and retardation of the DNA excision repair process. DFMO treatment did not alter the initial level of interaction of radiolabeled alkylating agent with cellular DNA, but combined treatment with DFMO and MGBG reduced covalent binding, probably through effects on cell cycling. Polyamine supplementation had no effects on initial yield of DNA single-strand breaks in drug-treated cells. The repair defect appeared similar to that observed previously in polyamine-depleted cells following X-irradiation and UV irradiation, namely retarded sealing of DNA strand breaks. It was not possible to reverse the effects of these inhibitors by short periods of polyamine loading, despite the fact that all three polyamines could be restored to near-normal levels. These findings provide the first demonstration of altered response of polyamine-depleted cells to monofunctional alkylating agents and contribute to our understanding of altered responses of polyamine-depleted cancer cells to a variety of DNA-reactive chemotherapeutic drugs. PMID- 8402581 TI - In vitro and in vivo tumoricidal properties of a pathogenic/free-living amoeba. AB - The chemotactic and tumoricidal properties of the pathogenic/free-living amoeba Acanthamoeba castellanii were examined in vivo and in vitro. A. castellanii trophozoites displayed strong positive chemotactic responses to human melanoma (OCM-1) and murine melanoma (D5.1G4) cells. Although the parasites typically invade and destroy the corneal epithelium and stroma, positive chemotactic responses were not detected against extracts of either of these corneal elements. In vitro studies revealed that viable parasites, as well as cell-free parasite lysates, produced swift and extensive cytolysis of a wide variety of tumor cells. The tumoricidal properties of A. castellanii were examined in vivo and revealed that injection of viable parasites or cell-free parasite lysates into progressively growing subcutaneous melanomas resulted in 83% and 53% reductions in the tumor masses compared with untreated controls. The feasibility of utilizing the tumoricidal properties of pathogenic/free-living amoebae and their cell-free products in the treatment of drug-resistant or radioresistant tumors warrants further investigation. PMID- 8402582 TI - Concentration-dependent effect of iron on gamma-linolenic acid toxicity in ZR-75 1 human breast tumor cells in culture. AB - Polyunsaturated fatty acids are cytotoxic to ZR-75-1 human breast tumor cells in culture. This effect may be potentiated by the simultaneous addition of iron. When cytotoxicity was measured in the presence of different concentrations of both gamma-linolenic acid and ferrous chloride there was an increase in cell death above concentrations of 9 microM and 0.05 microM, respectively. The potentiation of the effects of 18:3n-6 at low concentrations by the simultaneous addition of Fe(II) ions supports the contention that an alteration in the intracellular Fe(II)/Fe(III) ratio is necessary to promote autocatalytic lipid peroxidation. PMID- 8402583 TI - Squamous epithelial dysplasia associated with squamous cell carcinoma of the esophagus. AB - To investigate the relationship between dysplasia and carcinoma of the esophagus, 159 cases of esophageal carcinoma without any preoperative treatment were reviewed retrospectively. There were 75 dysplastic lesions in 32 cases (20.1%). The incidence of co-existence of dysplastic lesions was 0, 58.3, 31.3, 20.8 and 11.4% in intra-epithelial, mucosal and submucosal cancers and those invading the proper muscular layer and adventitia, respectively. Thus, excluding the cases of intra-epithelial carcinoma, the less advanced the lesion, the higher the incidence of dysplasia. Epithelial dysplastic lesions were classified as 12 with mild, 33 with moderate and 30 with severe degrees of dysplasia. Although the continuity of dysplastic lesions to the areas of carcinoma was not so frequent (48.0%), it was more often encountered in severe dysplasia rather than in moderate or mild dysplasia, which suggested some relationship between the severity of dysplasia and carcinoma. In the cases with a dysplastic lesion the multiplicity of squamous cell carcinoma and the intra-epithelial spread of the main lesion were more frequently seen (P < 0.001), suggesting a multicentric occurrence of dysplastic lesions and carcinomas. PMID- 8402584 TI - Inhibitory effects of semisynthetic flavonoid derivatives on the biochemical markers of tumor promotion in mouse epidermis in vivo. AB - Two sets of flavonoid derivatives were synthesized from condensed tannins (CTs) or catechin, and compared with the procyanidin monomer models, (+)-catechin and ( )-epicatechin, for their abilities to inhibit the biochemical effects of the potent tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in mouse epidermis in vivo. Topical applications of the semisynthetic flavonoids, catechin dialkyl ketals and epicatechin-4-alkylsulphides inhibit TPA-induced ornithine decarboxylase (ODC) activity to a much greater degree than catechin or epicatechin. Moreover, they reduce TPA-stimulated hydroperoxide (HPx) production, a response that cannot be inhibited by catechin or epicatechin. These compounds also inhibit the sequential stimulations of protein and DNA synthesis linked to TPA promotion. The remarkable effectiveness of these synthetic compounds, especially against the ODC marker of skin tumor promotion, suggests that they may be effective anti-tumor promoters. PMID- 8402585 TI - The extent and persistence of binding to respiratory mucosal DNA by inhaled tritiated propylene oxide. AB - In order to investigate some of the mechanisms underlying the carcinogenicity of inhaled propylene oxide (PO), the deposition and persistence of inhaled tritiated PO in the DNA of the nasal cavities, tracheae and lungs of rats were investigated. The results of dose/response exposure protocols revealed clear gradients for binding throughout the respiratory mucosa; the highest levels of binding were in the nasal mucosa and the lowest were in the lungs. Gradients became steeper as exposure concentrations were lowered. The persistence studies revealed that bound tritium declined in nasal mucosal DNA with apparent bi exponential kinetics while clearance from the tracheal and pulmonary DNA was much slower and occurred with apparent mono-exponential kinetics. Consequently, although the initial levels of binding of inhaled PO are lower in the trachea and lungs than in the nasal cavity, the slow clearance of PO from the DNA in these organs could increase the chances of a mutational event. PMID- 8402586 TI - Comparison of DNA adduct formation by means of synchronous scanning and by isotope tracers: in-vitro study on formation of DNA adducts in human lymphocytes exposed to benzo(a)pyrene. AB - The P-450 complex transforms polyaromatic hydrocarbons (PA) into active intermediates that may cause mutations due to DNA adduct formation. The present communication compares the rate of DNA adduct formation in human lymphocytes incubated for varying times with different concentrations of benzo(a)pyrene (B(A)P). In the presence of B(A)P the cultures were standardized as to medium and lectin concentration. Then the cells were exposed to increasing levels of cold and 3H-labelled B(A)P for varying times. B(A)P adducted to DNA was estimated both by synchronous scanning (SS) and by counting radioactivity. SS revealed a synchronous signal at 382 nm, corresponding to data available from the literature, and the peak height declined linearly with dose of B(A)P. However, it appeared that the signal height decreased if the DNA was successively extracted with chloroform. Five times extraction gave rise to a stable content of B(A)P of about 25% of the B(A)P originally found in the DNA. This DNA could only be traced with the radioactive tracer, since the concentration of adducted B(A)P was below the lower level of detection by the SS method. Even at low B(A)P levels the carcinogen exists in two forms in DNA: as 'free' non-adducted (extractable with lipid solvents) and an adducted form. Time variation showed that the DNA was incorporated linearly with both forms of B(A)P for up to 2 hours, then the take up was constant. Concentration variation showed linear incorporation up to 1 microM B(A)P. The present data may explain conflicting data on the extent to which lymphocytes adduct B(A)P to DNA. The lipid-soluble planar B(A)P of DNA may, like photosensitizers, intercalate between the two DNA strands. Like the intercalation of acridines into DNA, which is known to cause frameshift mutations, the intercalation of B(A)P may also have mutagenic consequences. PMID- 8402587 TI - A flow cytometric study of the rat Yoshida AH-130 ascites hepatoma. AB - The implantation of the Yoshida AH-130 ascites hepatoma to rats results in a marked reduction in body weight in the tumour-bearing hosts. This is associated with an important reduction in both food intake and energetic efficiency in the last period of tumour growth. The growth of the tumour mass has a clear initial exponential phase, in which the maximum cell density is reached, with the majority of cells being in the synthetic phase (S) followed by most of the cells being in the G0/G1 phase, as determined using flow cytometry. PMID- 8402588 TI - Changes of gamma/delta T cells in blood after radiation therapy for prostatic cancer. AB - The changes of the size of gamma/delta T (gamma delta T) and alpha/beta T (alpha beta T) cell subsets in the blood were examined after radiation therapy (66 Gy) for prostatic carcinoma in 17 men. Following radiation both subpopulations of T cells were reduced to 52% and 36% of starting values, respectively. gamma delta T cells were not replaced four months after radiation therapy, in contrast to alpha beta T cells, which were increased to 47% of initial values. The T alpha beta/T gamma delta ratio was significantly reduced immediately after radiation therapy but was normalised after four months. Blood gamma delta T cells may be reduced for a long time after radiation therapy and this may be true also for gamma delta T cells in irradiated epithelial organs and tumours. PMID- 8402589 TI - Reverse relationship between malignancy and cyclic AMP-dependent protein kinase activity in Yoshida rat ascites hepatomas. AB - Rat ascites hepatoma (AH) cells (10(6) cells/head) inoculated intraperitoneally into rats had host-killing ability (malignancy) in the order AH66F > AH44 > AH13 > AH7974 > AH109A > AH66 > AH130. The life span of the rats after inoculation closely correlated with the activity of cyclic AMP-dependent protein kinase (protein kinase A) in the tumor cells but not the activity of Ca2+/phospholipid dependent protein kinase (protein kinase C). N-[2-[N-[3-(4-chlorophenyl)-1-methyl 2-propenyl]amino]ethyl]-5- isoquinoline-sulfonamide (H-87), a potent, selective inhibitor of protein kinase A, inhibited in vitro growth of these hepatoma cells with a similar potency and, intraperitoneally injected, prolonged the lives of rats bearing less malignant AH66 cells (with high protein kinase A activity) but did not affect the life span of rats bearing highly malignant AH66F cells (with low protein kinase A activity). On the other hand N-(2-methylpiperazyl)-5 isoquinolinesulfonamide (H-7), an inhibitor of protein kinase C, inhibited AH66F cells more than AH66 cells, but did not influence the life span of rats bearing either hepatoma. From these results it is deduced that protein kinase A may be important in the regulation of malignancy and in vivo proliferation of AH cells. PMID- 8402590 TI - Coclastogenicity of ethanol with cigarette smoke in rat erythroblasts and anticlastogenicity in alveolar macrophages. AB - A series of experiments was carried out to assess cytotoxic and cytogenetic effects in bone marrow polychromatic erythrocytes (PCE) and pulmonary alveolar macrophages (PAM) resulting from individual or combined exposure of male BD6 rats to ethanol, cigarette smoke and Aroclor 1254. Addition of 5% ethanol to drinking water did not affect the micronucleus frequency but consistently enhanced the proportion of polynucleated PAM. Moreover, the higher concentration used (10%) was cytotoxic in the bone marrow. Whole-body exposure to cigarette smoke elevated the micronucleus frequency in both PCE (4.0-4.4-fold) and PAM (2.0-3.6-fold) and enhanced the frequency of polynucleated PAM. After 3 weeks of combined exposure, ethanol produced contrasting effects in smoke-exposed rats, i.e. an increase of micronuclei in PCE and a decrease in PAM. An i.p. injection of Aroclor 1254 was per se devoid of any influence on the monitored parameters but tended to attenuate the cytotoxic and cytogenetic changes produced by cigarette smoke or ethanol in both types of cell. PMID- 8402591 TI - Enhancement of cell killing of HL-60 cells by ultrasound in the presence of the photosensitizing drug Photofrin II. AB - Enhancement of acute cell killing of HL-60 cells by low-level ultrasound in the presence of the photosensitizing drug Photofrin II was examined. HL-60 cells were exposed to ultrasound (270 kHz) at intensities of 150, 300 and 450 mW/cm2 for 60 s in the presence of Photofrin II (200 micrograms/ml). Cell survival after treatment was 49.6 +/- 5.1%, 34.5 +/- 3.1% and 27.4 +/- 3.9%, respectively. Ultrasound exposure alone at 150, 300 and 450 mW/cm2 resulted in a decrease in cell survival to 92.9 +/- 1.5%, 82.3 +/- 2.2% and 77.9 +/- 7.2%, respectively. The cell survival rate after the addition of Photofrin II alone showed no significant cell killing. These results indicate that Photofrin II was sensitized by low-level, non-thermal ultrasound to enhance the cell killing of HL-60 cells. PMID- 8402592 TI - Dose response effects of 2-acetylaminofluorene on DNA damage, cytotoxicity, cell proliferation and neoplastic conversion in rat liver. AB - This study measured the effect of precise doses of 2-acetylaminofluorene (AAF) in inducing DNA damage, functional changes and neoplastic conversion in rat liver. Groups of male F344 rats at 9 weeks of age were exposed to cumulative doses of 0.5 or 2.0 mmol AAF per kg body weight given by gavage daily 5 days per week over an 8-week period and maintained with no further exposure for up to 8 weeks. Administration of AAF resulted in the formation of N-deoxyguanosin-(8-yl)-2 aminofluorene in liver DNA in relationship to dose. In centrilobular hepatocytes the zone of glutamine synthetase-expressing cells was reduced by exposure. By 8 weeks, but not at 4 weeks, the higher of the two doses of AAF provoked an increase in cell proliferation measured by immunohistochemical incorporation of bromodeoxyuridine. Altered hepatocellular foci expressing the placental form of glutathione transferase were induced by the high dose of AAF at 4 weeks, but not at the low dose. At 8 weeks the incidence of foci at the high dose was 79 times that induced by the low dose. These foci were highly proliferative. In animals exposed to AAF for 8 weeks and maintained for 4 weeks with no exposure, DNA adducts decreased by 80% and cell proliferation subsided by 80%, although the glutamine synthetase zone remained diminished. After discontinuation of AAF, the number of foci diminished by 50% and their proliferation subsided by 80% at 4 weeks, indicating a phenotypic reversion of many foci. With this protocol of administration of precise doses of AAF, we have established non-linearity of effects and a lack of correlation between DNA adduct formation and induction of cellular lesions. We suggest that doses in the range of those reported can be used to study the contribution of epigenetic and genotoxic effects in carcinogenesis and to study threshold events. PMID- 8402593 TI - Distinct cytotoxicity against neuroblastoma cells of peripheral blood and tumor infiltrating lymphocytes from patients with neuroblastoma. AB - The cytotoxicity against neuroblastoma cells of IL-2-activated peripheral blood (PBL) and tumor-infiltrating lymphocytes (TIL) was evaluated in seventeen patients with neuroblastoma. Regional lymph node lymphocytes (LNL) were similarly studied in some patients. Three allogeneic neuroblastoma cell lines, LA2D2, LA2B4 and SIFA, established from the different metastases of the same patient were used as targets. Of the three neuroblastoma lines, LA2D2, with low CD56 expression, was the most susceptible to IL-2-activated lymphocytes, while SIFA, with high CD56 expression, was resistant in the greatest degree. LA2B4 showed moderate susceptibility. Although TIL (73.9 +/- 2.1%), LNL (81.0%) and PBL (76.2 +/- 3.1%) revealed similar cytotoxic activity to K562, they demonstrated distinct cytotoxic activities to each neuroblastoma cell line, as follows: against LA2D2: TIL 56.3 +/- 4.2%, LNL 52.1%, PBL 33.6 +/- 4.9% (P < 0.01); against LA2B4: TIL 47.3 +/- 3.3%, LNL 37.8%, PBL 33.7 +/- 4.8% (P < 0.05); against SIFA: TIL 27.0 +/- 6.2%, LNL 20.7%, PBL 13.9 +/- 2.4% (P = 0.056). TIL always showed higher cytotoxic activity against neuroblastoma cells than those of LNL and PBL, whereas LNL were more cytotoxic than PBL. This data showed that TIL from neuroblastoma patients preferentially killed neuroblastoma cells. It was suggested that lymphocytes in the tumor site and regional lymph node could have been sensitized with neuroblastoma-related antigens and exert preferential killing activity against neuroblastoma cells. Phenotypical analysis revealed that TIL had a larger population of CD56+ cells than PBL. Conversely, PBL had a higher population of CD16+ cells than TIL. The cytotoxic activity of TIL significantly decreased by the depletion of CD56+ cells (10.9 +/- 6.2 from 49.9 +/- 5.9% against LA2D2, P < 0.001). These results indicated that CD56+ cells were most responsible for the killing of neuroblastoma cells, and that TIL, with a high proportion of CD56+ cells with strong activity, would be the best source for the immunotherapy of neuroblastoma. Additionally, since neuroblastoma cell lines used in the present study were derived from the different metastases of the same patient, heterogeneity in the susceptibility to lymphocytes might result from the differential expression of tumor-related antigens on these cell lines. PMID- 8402594 TI - Abnormal rectal cell proliferation and p52/p35 protein expression in patients with ulcerative colitis. AB - We evaluated the presence of cell proliferation and antigenic abnormalities in rectal biopsies from 37 patients affected by ulcerative colitis and 15 controls. The study was carried out by thymidine labeling and immunochemistry, using antibodies against specific cytoskeletal-associated proteins (p52, p35, alpha actinin). Among ulcerative colitis patients, 24 had an immunofluorescence pattern similar to that of controls, while 13 showed an abnormal distribution of one or more proteins (p52 alone or p52 and either p35 or alpha-actinin) within the rectal crypts. Patients showed a shift of the proliferative compartment towards the top of the rectal crypts compared with controls. This finding was more evident in patients with p52 or p35 abnormalities. Proliferative and antigenic defects were not related either to age or the duration of colitis. These phenotypic changes might be a biomarker of increased risk of colon cancer in ulcerative colitis. PMID- 8402595 TI - Increased ADP-ribosylation of histones in oral cancer. AB - The involvement of ADP-ribosylation has been investigated by comparing the extent of ADP-ribosylation in two different stages of oral cancer with that of the corresponding normal oral tissue. Poly(ADP-ribose) synthetase(PADPRS), the enzyme that is responsible for this type of posttranslational covalent modification, was assayed first by measuring 14C-ADP-ribose incorporation into different acceptor proteins of the tissue homogenate. The results clearly indicate an increase in PADPRS activity during oral carcinogenesis. After observing this increased pattern in total tissue, further experiments were conducted to check the extent of ADP-ribosylation in purified nuclei. The data of this experiment also indicates a progressive increase in the extent of ADP-ribosylation with increase in malignancy of oral tissue. Further analysis of an ADP-ribosylation of chromosomal proteins indicates that the increased pattern of ADP-ribosylation is mainly attributed to histones and not to non-histone chromosomal proteins. Our investigation suggests that ADP-ribosylation may play a role in oral cancer and may be used as a potential tumour marker. PMID- 8402596 TI - Effect of four plant phenols, beta-carotene and alpha-tocopherol on 3(H)benzopyrene-DNA interaction in vitro in the presence of rat and mouse liver postmitochondrial fraction. AB - The modulatory effect of beta-carotene, alpha-tocopherol and the four plant phenols eugenol, hydroxychavicol, curcumin and catechin on mouse and rat liver postmitochondrial fraction (S9 mix)-mediated 3(H)benzopyrene (B(a)P)-DNA interaction in vitro was studied. All the plant phenolics significantly inhibited 3(H)B(a)P-DNA interaction in the presence of both mouse and rat liver S9 fractions. In contrast, alpha-tocopherol proved to be ineffective in the presence of both types of S9 mix, whereas beta-carotene inhibited only mouse S9 mix mediated 3(H)B(a)P-DNA interaction. PMID- 8402597 TI - Correlation of the ability of retinoids to inhibit promoter-induced anchorage independent growth of JB6 mouse epidermal cells with their activation of retinoic acid receptor gamma. AB - Retinoids inhibit the biological effects induced in mouse epidermal cells by the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Specific nuclear retinoic acid receptors (RARs) have been identified in the epidermis, but the specific receptor that mediates the inhibitory response by retinoids is not established. Retinoic acid and six conformationally restricted retinoids were evaluated in an in vitro bioassay using the JB6 mouse epidermal cell line. These activities were then compared with the ability of these retinoids to activate the RARs in transient transfection assays for transcriptional activation to identify the retinoid receptor involved in inhibiting TPA-induced anchorage-independent growth. The retinoids inhibited TPA-induced colony formation of JB6 cells in semisolid medium at concentrations that were not toxic based on colony formation of attached cells. These concentrations ranged from less than 10(-9)-10(-6) M. 4 (5,6,7,8-tetrahydro-5,5,8,8-tetramethylanthracen-2-yl)benzoic acid (TTAB) was the most potent retinoid, with an EC50 of 0.8 nM. Both RAR alpha and RAR gamma were expressed in JB6 cells. Expression of RAR beta was not detected in these cells using a polymerase chain reaction assay, consistent with its extremely low level in mouse skin. Inhibition of the TPA response by these retinoids in JB6 cells correlated only with their transcriptional activation of RAR alpha, but not with that of RAR alpha. These results suggest that RAR gamma is most probably the receptor that mediates the chemopreventive effects of retinoids in mouse epidermis. PMID- 8402598 TI - Germ-line p53 mutation is uncommon in patients with triple primary cancers. AB - We examined five patients with multiple primary cancers who had a history of three different types of primary cancers for germ-line p53 mutations. The germ line p53 mutation was detected in a patient who conformed to the Li-Fraumeni syndrome, but not in the other four patients. The diagnosis of these four patients did not fall in the category of Li-Fraumeni syndrome. This result indicates that germ-line p53 mutations are uncommon even in patients with triple primary cancers. PMID- 8402599 TI - Pharmacokinetic and tumour-photosensitizing properties of the cationic porphyrin meso-tetra(4N-methylpyridyl)porphine. AB - The pharmacokinetic behaviour and the photodynamic properties of the cationic porphyrin meso-tetra(4N-methylpyridyl)porphine (T4MPyP 2.1 mg/kg) were examined in Balb/c mice bearing an MS-2 fibrosarcoma. The porphyrin shows good tumour localizing properties; 24 h after drug administration the tumour concentration of T4MPyP was approximately 1.2 ng/mg, while the concentrations in normal tissues were substantially lower, except for liver and spleen. In the serum, T4MPyP is preferentially transported by albumin and globulins (80.5%), while minor amounts are associated to lipoproteins (19.5%). The phototherapeutic efficiency of T4MPyP was tested by following the growth curves of fibrosarcoma irradiated by 600-680 nm (450 J/cm2) 24 h after the i.v. injection of T4MPyP (2.1 mg/kg). PDT-treated tumours showed a temporary delay in their growth compared with control tumours. The excellent selectivity of T4MPyP and its antitumour activity on photoexcitation encourage further studies for assessing the usefulness of this porphyrin in photodynamic therapy. PMID- 8402600 TI - Effects of eicosanoid metabolism inhibitors on growth of a human gastric tumour cell line (HGT). AB - HGT cells are a human gastric cell line derived from a tumour of the stomach. We have investigated the effects of lipoxygenase and cyclooxygenase metabolism inhibitors on HGT cell proliferation, on fatty acid composition of HGT cells and on the incorporation and distribution of arachidonic acid (AA) in HGT lipids. The cyclooxygenase inhibitor aspirin and both cyclooxygenase and the lipoxygenase inhibitor BW 755C suppressed cell proliferation in a concentration-dependent manner. The inhibition of HGT proliferation did not result from a modulation of the fatty acid composition of membrane lipid, which was not affected by treatment with the various inhibitors. Inhibitors of AA metabolism did not alter acylation of exogenous AA into HGT cells nor its subsequent distribution in the lipid and phospholipid species. The role of cyclooxygenase eicosanoids in HGT proliferation is discussed. PMID- 8402601 TI - Relationships of barriers and facilitators to breast self-examination, mammography, and clinical breast examination in a worksite population. AB - The American Cancer Society recommends a regimen for breast cancer screening that includes mammograms, clinical breast examination, and breast self-examination. Compliance with breast cancer screening guidelines has been linked to a number of barriers and facilitators. These barriers and facilitators seem to lie within the cognitive framework and generalized beliefs of women, and in the situational contexts in which they lead their lives. A comprehensive study was designed to investigate variables related to breast cancer screening behaviors (breast self examination, mammography, and clinical breast examination) of working women > or = 35 years of age at their worksite environments. A factor analysis identified similar sets of composite variables related to each of the screening modalities, and a discriminant analysis was performed for each screening technique to identify those variables that were most significant in predicting compliance with screening guidelines. The variables discomfort, perceived efficacy, and desire for control over health were significant for all three screening behaviors. Perceived importance was identified as a fourth variable for mammography and clinical breast examination, and lack of knowledge was a fourth variable for breast self-examination. Effective breast cancer screening programs involve all three screening techniques. In the design of education and intervention programs at worksites, it is critical to emphasize the commonalities of the variables that emerged in this study as important for each screening technique. Health-care professionals who implement such intervention programs need to explore and bring into the open these common barriers and facilitators to maximize working women's compliance with breast screening guidelines. PMID- 8402602 TI - Mammography in women > or = 50 years of age. Predisposing and enabling characteristics. AB - The purpose of this study was to identify the relationship of selected predisposing and enabling characteristics of women > or = 50 years of age to mammography utilization. Andersen and Aday's theoretical model for health services utilization guided data collection. Data were collected from a convenience sample of 161 women members of four urban churches, using a mailed survey. Results showed that 81% reported at least one mammogram and 24% had followed mammography guidelines for the preceding 3 years. Results of logistic regression analyses with variables having a bivariate significance of p < or = 0.01 showed that higher income was associated with both ever having a mammogram and adherence. Willingness to pay > $50 out of pocket for a mammogram was significant for 3 year adherence. Additionally, the sociodemographic variables of age and religion were associated with adherence, whereas a college education was highly significant (odds ratio = 13.78) for ever having a mammogram. Having a regular place for health care and having yearly Papanicolaou tests were associated with ever having a mammogram, but not adherence. Finally, intending to get a mammogram was associated with ever having a mammogram. In this study, belief and knowledge variables showed no association with utilization, and social influence had bivariate significance only for ever having a mammogram. This study suggests the importance of addressing economic and health-care delivery system factors to promote increased mammography utilization, particularly for older women. PMID- 8402603 TI - Who cares? Patients' descriptions of age-related aspects of cancer and care in Stockholm. AB - This exploratory study examines the situation of a heterogeneous group of 46 cancer patients from one general hospital in the Stockholm area who were diagnosed with a malignant disease in 1987. Forty-six patients were interviewed to determine how they experience and cope with their sickness. Although age related issues were not directly addressed by the interviewer, they were frequently commented upon by the respondents. A questionnaire was also used to quantify various psychosocial aspects of the individual's cancer experience. Quantitative and qualitative data were used in a complementary fashion. The patients interviewed ranged in age from 29 to 88 years (median 61). Issues related to age were frequently addressed spontaneously by the interviewed persons, which led to further exploration of age-related aspects of care. Although few age-related differences in symptom distress were found, some differences were seen in patients' perceptions of the response of the professional health-care system. The older patients related that they have less quantitative contact with specialized formal resources, and perceive less sense of engagement and concern from the professional health-care sector. The qualitative analysis suggests that age may be used by patients as an explanatory factor in a variety of situations, functioning as a means of "making sense" of sickness experiences. PMID- 8402604 TI - Percutaneous hepatic arterial chemotherapy and chemoembolization. AB - Percutaneous hepatic arterial chemotherapy and chemoembolization are treatment options for patients who have been diagnosed with hepatocellular carcinomas or with metastatic carcinomas of the liver. Nursing care of these patients is challenging and complex. Patients present with varying symptoms of the disease, and progression requires varying degrees of intervention. Some patients may have already had systemic chemotherapy, and others may be facing their first experience with chemotherapy. Patient and family education requires presentation of correct, thorough information on an ongoing basis. To care for these patients, the nurse must understand the techniques of chemoembolization and hepatic arterial chemotherapy administration. This article addresses the introduction of hepatic arterial chemotherapy through the process of chemoembolization, as well as the nursing management to be considered throughout the treatment. PMID- 8402605 TI - Acute symptoms associated with antineoplastic drug handling among nurses. AB - Antineoplastic drug handling in the absence of adequate protective measures has been associated with biological uptake of the drugs among pharmacists and nurses. This study investigated the association between occupational exposure to antineoplastics and the presence of acute symptoms in a nationwide sample of 2,048 nurses and nurses' aides. Reported skin contact with the drugs was associated with a small but statistically significant increase in reported symptoms. Although number of doses handled and extent of protection used were significantly associated with number of symptoms, their effect was not independent of skin contact. PMID- 8402606 TI - Nursing strategy for initial emotional response to cancer diagnosis. AB - Nursing strategy for patients' initial emotional response to cancer was conducted with five clients and evaluated for effectiveness using a case study approach. This clinical evaluation of a management strategy was conducted in an acute-care setting during the phase of initiating a therapeutic relationship. The nurse identified stressors, support systems, usual coping strategies, and level of knowledge about the disease. Two meetings a week were conducted with each client during hospitalization. Evaluation was completed on four of the five clients. The management strategy facilitated beginning reintegration of body, mind, and spirit resulting in a smoother transition phase. The aim of this protocol was to initiate effective coping responses to a potentially devastating diagnosis. PMID- 8402607 TI - The meaning and impact of empathic relationships in hospice nursing. AB - This naturalistic field study was designed to explore the patient's perspective of the nature, meaning, and impact of empathic relationships with hospice nurses. The findings are part of a larger study, focused on the meaning and impact of empathic relationships that develop between hospice nurses and their patients. Data were generated through in-depth interviews with 14 terminally ill adults receiving home-based hospice care. According to the hospice patient, an empathic relationship developed through a process of reciprocal sharing and revealing of personhood within a context of caring and acceptance. The experience of an empathic relationship meant being acknowledged as an individual, a person of value. The outcome of the empathic relationships between hospice nurses and their patients was the improvement and maintenance of patients' physical and emotional well-being. Understanding the patient's perspective is critical for effective nursing interventions and meaningful outcomes. Future research needs to explore empathic relationships between the nurse and family caregivers in various settings. PMID- 8402608 TI - Chemotherapy agents: Part I. PMID- 8402609 TI - The small heat shock protein hsp27 is correlated with growth and drug resistance in human breast cancer cell lines. AB - An emerging body of evidence suggests that the heat shock proteins (hsp) may be involved in drug resistance. When hsp are induced by elevated temperatures, resistance to doxorubicin (Dox), but not to other commonly used chemotherapeutic agents, is induced in breast cancer cells. To evaluate the role of hsp27 in this phenomenon, we have transfected MDA-MB-231 breast cancer cells, which normally express low levels of hsp27, with a full-length hsp27 construct. These hsp27 overexpressing cells now display a 3-fold elevated resistance to Dox. Anchorage dependent proliferation and anchorage-independent growth were also increased 2-4 fold in these transfectants. We have also derived a MCF-7 breast cancer cell line with amplified endogenous hsp27 which is highly resistant to Dox. When these cells are transfected with an antisense hsp27 construct, they are rendered sensitive to Dox (3-fold) with anchorage-dependent as well as anchorage independent growth, similarly decreased. These results suggest that hsp27 specifically confers Dox resistance in human breast cancer cells and, furthermore, that hsp27 may be involved in the regulation of cell growth. PMID- 8402610 TI - Detection of the AML1/ETO fusion transcript in the t(8;21) masked translocation in acute myelogenous leukemia. AB - The fusion transcript AML1/ETO was detected in the bone marrow of two t(8;21) negative acute myelogenous leukemia (AML) patients by means of reverse transcription-polymerase chain reaction. This fusion transcript is identical to the one transcribed from the t(8;21) translocation base, as deduced from (a) the size and restriction pattern of the amplified DNA fragment and (b) the DNA sequence analysis of the fusion junction. We also showed that the ETO gene is highly expressed in these patients, much as it is in the t(8;21)-positive AML. Southern blot analysis showed rearrangement of the AML1 gene in one of the patients. Together, our results demonstrate that there is a masked t(8;21) translocation in AML that is not detectable by cytogenetic analysis but is able to transcribe an AML1/ETO fusion transcript similar to that transcribed in t(8;21)-positive AML-M2 patients. PMID- 8402611 TI - Dominant negative effect of a germ-line mutant p53: a step fostering tumorigenesis. AB - Lysates derived from the fibroblasts of individuals who are homozygous for normal p53 or heterozygous for the germ-line p53 mutation characteristic of certain Li Fraumeni cancer-prone families were assessed for p53 function utilizing the binding of p53 protein to a p53-specific consensus oligonucleotide sequence. As expected, control nuclear lysates containing only mutant p53 or no p53 displayed little or no such binding. However, the nuclear lysates from heterozygous fibroblasts containing similar amounts of normal p53 and 245D mutant p53 displayed binding that was significantly below 50% of that seen with homozygous wild-type p53 in normal cell lysates. The nuclear lysates of these heterozygous or homozygous fibroblasts exhibited similar levels of DNA binding to a consensus oligonucleotide specific for the transcription factor, AP-1. These results indicate that mutant p53 has a transdominant effect on the binding of DNA by normal p53. These findings also suggest that p53 complexes formed in vivo that contain mutant p53 are functionally impaired even if normal p53 is also present in the complex. The implications of a trans-dominant effect of mutant p53 on the cancer-prone phenotype of individuals heterozygous for mutated p53 in Li-Fraumeni families is discussed. PMID- 8402612 TI - Comparison of loss of heterozygosity patterns in invasive low-grade and high grade epithelial ovarian carcinomas. AB - Loss of heterozygosity (LOH) studies were performed to investigate the genetic differences which separate low-grade (LG), high-grade (HG), and borderline epithelial ovarian carcinomas. Fresh tumor samples and blood were obtained from 58 patients (20 LG, 34 HG, and 4 borderline tumor specimens) undergoing surgery for ovarian carcinoma at Mayo Clinic. Tumors were graded using a modified Broder's classification with invasive grades 1 and 2 considered LG, invasive grades 3 and 4 considered HG, and tumors with no evidence of stromal invasion classified as borderline. Polymorphism analysis was performed using 76 restriction fragment length polymorphisms and variable number of tandem repeats and 59 microsatellite markers representing all chromosome arms. Chromosome arms 6p, 17p, 17q, and 22q were found to be frequently lost in LG as well as HG tumors. Chromosome arms 13q and 15q were lost to a significantly greater extent in HG tumors compared to LG neoplasms (P = 0.003 and P = 0.08, respectively). Conversely, 3p loss was seen more frequently with LG tumors (P = 0.02). The majority of LG tumors (65%) did not show frequent LOH in the allelotype analysis. In fact, a subset of 7 (7 of 20) LG tumors accounted for 76% of the total allelic loss in the LG category. These tumors showed LOH almost identical to that of the HG neoplasms. Borderline tumors showed a low rate of allelic loss. There were no common events found between borderline and invasive tumors. Our data suggest that most HG tumors and a subset of LG tumors share genetic alterations at putative tumor suppressor genes detected by LOH studies. Chromosome 6 and 17 losses appear to be early events while 13q and 15q losses appear to be critical late events. However, a majority of LG tumors appear to develop as a consequence of an alternative mechanism(s) which is not detected by LOH studies. Possibilities include: (a) inactivation of tumor suppressor genes without LOH; (b) dominant negative gene(s) in which only one allele requires mutation; and (c) changes in dominant acting oncogenes. This unidentified phenomenon may be operative in borderline tumors as well. PMID- 8402613 TI - Dietary fenretinide, a synthetic retinoid, decreases the tumor incidence and the tumor mass of ras+myc-induced carcinomas in the mouse prostate reconstitution model system. AB - Several epidemiological studies have implicated low dietary and serum levels of retinol with an increased risk for the development of human prostate cancer. In a recent report, dietary fenretinide [N-[(4-hydroxyphenyl)] retinamide], a synthetic retinoid with low toxicity, decreased the incidence of experimentally induced prostate cancer. Fenretinide is currently being evaluated in phase I and phase II clinical trials as an agent for both the treatment and chemoprevention of human prostate cancer. Because of these findings, we investigated whether dietary fenretinide could alter the incidence of phenotype of oncogene-induced prostate cancer in the mouse prostate reconstitution model system. When compared to control-fed animals, dietary fenretinide reduced the tumor incidence by 49% and the tumor mass by 52% of ras+myc-induced cancers in the mouse prostate reconstitution model system, which was modified to prolong the latency period before cancer development. Retinoids have a wide ranging effect on cellular differentiation, growth factor synthesis, and immune function. While its mechanism of action in this system remains unclear, fenretinide is an effective agent for the chemoprevention and growth modulation of oncogene-induced prostate cancer in the mouse prostate reconstitution model system and may be effective for the chemoprevention of human prostate cancer. PMID- 8402614 TI - Induction of protein-DNA complexes in HeLa S3 cells by KW-2149, a new derivative of mitomycin C. AB - KW-2149 is a new water-soluble analogue of mitomycin C which shows a broad spectrum antitumor activity in experimental tumor models including mitomycin C resistant tumors and is less hematotoxic than mitomycin C. We have found that KW 2149 increases protein-DNA complexes in HeLa S3 cells using the potassium/sodium dodecyl sulfate precipitation method which recovers largely DNA bound covalently to protein but does not precipitate DNA bound noncovalently to proteins. In contrast, no increase of the protein-DNA complexes are shown in cells treated with mitomycin C. Thus, KW-2149 binds covalently with both DNA and protein, and this may mediate for KW-2149 the potent and characteristic antitumor activity compared with mitomycin C. PMID- 8402615 TI - Redox modulation of p53 conformation and sequence-specific DNA binding in vitro. AB - The p53 protein is a transcription factor, the function of which is abrogated by oncogenic mutations which affect a flexible domain in the central portion of p53, altering its reactivity with conformation-specific antibodies. Here we show that both conformation and sequence-specific DNA binding of p53 translated in vitro can be modulated by metal chelators and oxidizing agents. Oxidation disrupted wild-type p53 conformation and inhibited DNA binding. Conversely, reduction favored folding of p53 into the wild-type form and restored DNA binding. Redox regulation of p53 protein conformation could represent an important mechanism for the control of p53 function. PMID- 8402616 TI - Activation of phospholipase D participates in signal transduction pathways responsive to gamma-radiation. AB - Early responses of mammalian cells to ionizing radiation include the activation of a protein kinase C implicated in the regulation of gene expression, the stimulation of tyrosine kinase activities, and the enhancement of phosphatidylinositol turnover. In the present report we show that clinically relevant doses of gamma-radiation (2 Gy) stimulate phosphatidylcholine hydrolysis in human squamous carcinoma cells. Radiation induced the accumulation of intracellular [3H]choline and the simultaneous increase in [3H]myristoyl phosphatidic acid, followed by a small increase in the levels of [3H]myristoyl diacylglycerol. Furthermore, in the presence of ethanol, gamma-radiation stimulated the appearance of [32P]phosphatidylethanol, an indicator of phospholipase D transphosphatidylation activity. These data demonstrate for the first time that phospholipase D activation participates in signaling pathways in response to gamma-radiation. PMID- 8402617 TI - The incidence of p53 mutations increases with progression of head and neck cancer. AB - To establish a genetic model of the progression of head and neck squamous carcinoma we have defined the incidence and timing of p53 mutations in this type of cancer. We sequenced the conserved regions of the p53 gene in 102 head and neck squamous carcinoma lesions. These included 65 primary invasive carcinomas and 37 noninvasive archival specimens consisting of 13 severe dysplasias and 24 carcinoma in situ lesions. The incidence of p53 mutations in noninvasive lesions was 19% (7/37) and increased to 43% (28/65) in invasive carcinomas. These data suggest that p53 mutations can precede invasion in primary head and neck cancer. Furthermore, the spectrum of codon hotspots is similar to that seen in squamous carcinoma of the lung and 64% of mutations are at G nucleotides, implicating carcinogens from tobacco smoke in the etiology of head and neck squamous carcinoma. PMID- 8402618 TI - Human immunodeficiency virus type 1 Tat protein protects lymphoid, epithelial, and neuronal cell lines from death by apoptosis. AB - We report here that the tat gene product of human immunodeficiency virus type 1 was able to protect lymphoblastoid (Jurkat), epithelial (293) and neuronal (PC12) cell lines from apoptotic death induced by serum withdrawal. The rescue from apoptosis by Tat was reflected by an increased expression of Bcl-2 protein in tat positive Jurkat cells with respect to mock-transfected Jurkat cells after 3-6 days of serum-free cultures. We propose that the ability of the regulatory human immunodeficiency virus type 1 Tat protein to suppress apoptosis might have important implications in understanding the pathogenesis of frequent neoplastic disorders observed in human immunodeficiency virus type 1-seropositive individuals. PMID- 8402619 TI - Refinement of regional loss of heterozygosity for chromosome 11p15.5 in human breast tumors. AB - The familial association of breast cancer with other tumors such as rhabdomyosarcoma that show loss of heterozygosity (LOH) for chromosome 11p15 as well as limited analyses showing LOH for chromosome 11p in breast tumors suggests the presence of a pleiotropic tumor suppressor gene in this region. In order to test this idea, we analyzed DNA samples for 50 matched normal and tumor tissues from unselected breast cancer patients for LOH at loci throughout the chromosome 11p15.5 region. We found that 12.5% of informative cases showed LOH at HRAS1, 26.8% at TH, and 33.3% at both D11S860 and HBB, providing genetic support for this hypothesis. In contrast to previous observations which excluded the involvement of 11p15.5 regions distal to the HBB cluster, our results indicate that the subregion between TH and HBB is a critical region in breast cancer. This region is identical to that identified for the clinically associated tumor, rhabdomyosarcoma, and thus warrants intensive molecular analysis. PMID- 8402620 TI - Potential topoisomerase II DNA-binding sites at the breakpoints of a t(9;11) chromosome translocation in acute myeloid leukemia. AB - We have examined a t(9;11)(p22;q23) chromosome translocation in an acute myeloid leukemia of an infant. The breakpoints on the two chromosomes occurred within introns of the involved genes: AF-9 on chromosome 9, and ALL-1 on chromosome 11. Sequence analysis identified heptamers flanking the breakpoints on both chromosomes 9 and 11, suggesting that the V-D-J recombinase was involved in the translocation. The presence of an N-region between the two chromosomes supports the hypothesis that a mistake in V-D-J joining was involved in the genesis of the translocation and indicates that terminal deoxynucleotidyl transferase was expressed in the cells from which this acute myeloid leukemia originated. In addition, potential topoisomerase II DNA-binding sites were found near the breakpoints of both chromosomes, suggesting the involvement of altered topoisomerase II activity in this translocation. Altered topoisomerase II activity in the presence of an active V-D-J recombinase may be a pathogenetic mechanism of acute myeloid leukemia with rearrangements at 11q23. PMID- 8402621 TI - Ability of aberrant crypt foci characteristics to predict colonic tumor incidence in rats fed cholic acid. AB - Aberrant crypt foci (ACF) are putative preneoplastic lesions of colon cancer which are being utilized currently as a biological end point to evaluate the induction and modulation of colon carcinogenesis. In several previous short-term studies, the unexpected reduction of ACF by the reported colonic tumor promoter cholic acid (CHA) emphasized the need for a systematic evaluation of the growth of ACF in response to a tumor promoter. The present study was conducted to determine if any characteristic(s) of ACF at various early stages of carcinogenesis would predict resulting tumor incidence in rats fed CHA. Male Sprague-Dawley rats received two injections of azoxymethane (20 mg/kg) and were fed either the AIN-76 diet or AIN-76 plus 0.2% CHA. The number, crypt multiplicity (number of crypts/focus), and size (area) of ACF were measured after 2, 8, 14, and 18 weeks in 5 rats/group. The number of ACF was lower (P < 0.033) in animals fed CHA at all time points. Average crypt multiplicity of ACF was greater (P = 0.045) from CHA-fed animals after 8 weeks compared to animals fed the AIN-76 diet. The average size of ACF was smaller in CHA-fed animals after 2 weeks and then tended to be larger than the sizes of the ACF from animals fed the AIN-76 diet. All remaining animals were killed after 18 weeks. Tumor incidence was higher (P < 0.001) in the CHA-fed group (63.2%) compared to the control diet group (29.4%). CHA-fed rats also had a higher number of tumors/tumor-bearing rat compared to control diet rats (1.96 versus 1.13). The main finding of this study is that the number of ACF at early time points did not predict tumor incidence. Crypt multiplicity was a consistent predictor of tumor outcome and should be measured in future studies using ACF as a biological end point. The CHA diet appears to provide a unique tumor-modulating environment that selectively enhances the growth of a smaller number of ACF leading to an increased number of tumors compared to a control diet. The mechanism(s) by which CHA mediates this effect warrants further investigation. PMID- 8402622 TI - Human papillomavirus 16 immortalization of normal human ectocervical epithelial cells alters retinoic acid regulation of cell growth and epidermal growth factor receptor expression. AB - Retinoids are potent regulators of epithelial cell growth and differentiation. Recently, they have been demonstrated to be effective in the treatment of preneoplastic cervical lesions in which human papillomavirus (HPV) is expressed. To better understand the mechanism of the antineoplastic effect of retinoic acid on HPV-positive cells, the effects of retinoic acid on both normal and HPV immortalized human ectocervical epithelial cell growth, epidermal growth factor (EGF) receptor level, and EGF receptor function were investigated. Both HPV immortalized cells (ECE16-1) and normal ectocervical cells (ECE cells) are growth stimulated by EGF. ECE16-1 but not normal ectocervical epithelial cells are growth inhibited by trans-retinoic acid which attenuates the stimulatory effect of EGF on ECE16-1 cell growth. Retinoic acid reduces both EGF binding and EGF receptor protein levels in ECE16-1 cells but not in normal ectocervical cells. The reduction in EGF receptor binding and receptor protein levels in ECE16-1 cells is not associated with the induced secretion of a soluble EGF receptor ligand, altered EGF receptor affinity, receptor internalization, or decreased receptor stability. Interestingly, the level of EGF receptors is consistently elevated in the ECE16-1 cell line as compared to normal ectocervical epithelial cells. Investigation of a second HPV-immortalized cell line (ECE16-D1) and two other HPV-positive cervical carcinoma cell lines revealed similar elevated EGF binding capacity and regulation by retinoic acid. In contrast, two HPV-negative cervical carcinoma cell lines demonstrated various EGF-binding levels but demonstrated no significant loss of EGF binding following retinoic acid treatment. Other normal cells and an SV40 large T-antigen-immortalized foreskin keratinocyte cell line, KER-1, had EGF receptor levels similar to the normal ectocervical epithelial cells, and no regulation by retinoic acid was observed. These data indicate that HPV immortalization may increase EGF receptor levels in ectocervical cells, elevating their sensitivity to growth stimulation by EGF, and that retinoic acid can possibly attenuate this increased responsiveness to EGF. PMID- 8402623 TI - Chronic radiation-induced alteration in hematopoietic repair during preclinical phases of aplastic anemia and myeloproliferative disease: assessing unscheduled DNA synthesis responses. AB - Protracted, low-daily-dose gamma-ray exposure (3.8-7.5 cGy/day) segregates canines into separate survival- and pathology-based subgroups by the early elicitation of distinct, repair-mediated hemopathological response pathways. In this study, we verified the blood and marrow responses of two major subgroups prone to either aplastic anemia or myeloproliferative disease, along with two variants, and extended our analyses of hematopoietic repair to include studies of DNA repair in bone marrow blasts using an autoradiographically based unscheduled DNA synthesis (UDS) assay. The myeloproliferative disease-prone subgroup exhibited extended survival (> 200 days), related to partial, gradual restoration of blood leukocyte, platelet, and marrow progenitor levels following an initial phase of acute suppression. Marrow blasts taken during the restoration phase showed expanded and qualitatively modified UDS relative to marrow blasts of age matched control animals. The amount of UDS per blast (signal strength) increased significantly, as did the number of UDS-positive cells and their sensitivities to high-dose UV induction and 1-beta-D-arabinofuranosylcytosine chemical inhibition. A nonevolving myeloproliferative disease-prone variant having prolonged survival (> 200 days) and restored blood cells and marrow progenitor levels also had marrow blasts with expanded UDS responses, but these were uniquely evoked by low (but not high) doses of UV inducer. The aplastic anemia-prone subgroup was characterized by short survival (< 200 days), progressive decline (without restoration) in all measured blood and marrow compartments, and largely nonsignificant changes in UDS responses of marrow blasts. A variant of this aplastic anemia-prone subgroup (with comparable short survival due to markedly ineffective hematopoiesis, but expressing select preleukemic features) exhibited reduced numbers (relative to age-matched controls) of highly responsive, UDS positive marrow blasts (in terms of UDS signal strength and increased to sensitivity 1-beta-D-arabinofuranosylcytosine-induced UDS inhibition). From these observations we conclude that: (a) the UDS response of marrow blasts, a correlate of hematopoietic progenitorial repair, is altered differentially within selected subgroups of animals under chronic radiation exposure; and (b) the nature of altered UDS repair response patterns appears to be largely related to the preclinical status/predisposition of the individual animal and thus may provide prognostically useful information in the clinical monitoring of chronically irradiated individuals with minimal but evolving hematological disease. PMID- 8402624 TI - Major difference in the hepatocarcinogenicity and DNA adduct forming ability between toremifene and tamoxifen in female Crl:CD(BR) rats. AB - The hepatoproliferative effects of 2 antiestrogens, tamoxifen and toremifene, were compared in a sequential 15-month study in which 2 doses of each compound were administered by daily gavage to female Sprague-Dawley rats for up to 12 months. The doses were 11.3 and 22.6 mg/kg for tamoxifen and 12 and 24 mg/kg for toremifene. There were scheduled sacrifices at 3, 6, 12, and 15 months, the latter including a 3-month recovery period from the 12th through the 14th month. In the chronic toxicity study, tamoxifen at 22.6 mg/kg produced 100% incidence of hepatocellular carcinoma at the 12- and 15-month sacrifice intervals and 67% and 71% incidences at the 11.3-mg/kg dose. Sequential observations showed an increased incidence of glutathione S-transferase-positive foci of hepatocellular alteration by 3 months with tamoxifen in the absence of hepatotoxicity, with the first liver carcinoma appearing by 6 months of treatment. Unscheduled deaths occurring beyond 7.5 months in the tamoxifen treated groups were due in almost all cases to liver cancer. In striking contrast, toremifene did not produce any hepatoproliferative effects at 12- and 24-mg/kg dose levels, nor in a pilot study at 48 mg/kg. The 24-mg/kg dose of toremifene exerted an inhibiting effect on foci of hepatocellular alteration in rat liver detectable by glutathione S-transferase immunohistochemistry at 3 months and by conventional histology at 12 months. An antiproliferative effect was also evident in mammary gland and anterior pituitary where both toremifene and tamoxifen suppressed tumor incidence in comparison to the control group. The ability of these drugs to modify rat liver DNA after p.o. administration was investigated using the 32P-postlabeling assay. Administration of tamoxifen at 45 mg/kg for 7 days produced liver DNA nucleoside modifications represented by 7 spots on the autoradiogram. Unlike tamoxifen, toremifene did not produce any modified bases in rat liver DNA detectable by the 32P-postlabeling technique. The dose levels of tamoxifen that are strongly hepatocarcinogenic in the rat are compared with doses used in humans in various applications. Taking internal drug exposure into account, we conclude that the margin of safety for use of tamoxifen as an endocrine prophylactic agent for healthy, but breast cancer prone, women is questionable. PMID- 8402625 TI - Regulation of protein kinase C isozymes in kidney regeneration. AB - Tissue damage and repair processes are important factors in renal tumor progression. To determine whether protein kinase C (PKC) is involved in these processes, we characterized PKC isozymes during rat kidney regeneration using 3 models: (a) diffuse cortical hyperplasia and hypertrophy induced by folic acid; (b) focal necrosis of the S3 segments induced by S-(1,2-dichlorovinyl)-L cysteine; and (c) compensatory renal hypertrophy. Immunoblot analyses demonstrated that 5 PKC isozymes, alpha, beta, delta, epsilon, and zeta, were expressed in rat kidney. Six h after folic acid treatment, phorbol ester receptors were down-modulated. Down-modulation preceded an increase in DNA synthesis which was maximal at 24 h. The reduction in phorbol ester receptors was due largely to a decrease in alpha-PKC. zeta-PKC, which is not a phorbol ester receptor, was also decreased. delta- and epsilon-PKCs were not changed. However, alpha-PKC was not down-modulated during compensatory hypertrophy induced by unilateral nephrectomy. Thus, the observed decrease of alpha-PKC after folic acid treatment is most likely associated with the hyperplastic and not the hypertrophic effects of this renal toxin. These results demonstrate that activation-associated down-modulation of PKC, in particular alpha-PKC, occurs during chemical-induced renal regeneration and suggests a general role for PKC activation in non-phorbol ester tumor promotion. PMID- 8402626 TI - Expression and prognostic significance of platelet-derived growth factor and its receptors in epithelial ovarian neoplasms. AB - The expression of platelet-derived growth factor (PDGF), PDGF-alpha receptor (PDGFR alpha) and PDGF-beta receptor (PDGFR beta) was studied in normal ovaries and ovarian neoplasms by immunohistochemical analysis. PDGF was detected in tumor cells in 33 of 45 malignant tumor samples but in none of 20 benign tumors (P < 0.001) or 11 normal ovaries (P < 0.001). In borderline tumors, 4 of 7 tissues stained positive in tumor cells. PDGFR alpha was detected in tumor cells in 16 of 45 malignant tumors, while no epithelial staining was found in 16 benign tumors (P = 0.002) or in 10 normal ovaries (P = 0.023). In 1 of 7 borderline neoplasms, tumor cells expressed PDGFR alpha. Neither normal epithelium nor tumor cells stained positive with antibodies against PDGFR beta. Patients with ovarian cancer and PDGFR alpha-positive tumor cells demonstrated an overall shorter survival compared to those who had negatively stained tumors (P < 0.005). A similar correlation was found in patients having stage III ovarian cancer (P < 0.01), which further supports an independent role for PDGFR alpha as a prognostic factor. Thus, the concomitant expression of PDGF and PDGFR alpha in tumor cells is related to progression of malignant ovarian tumors, indicating a functional role of PDGF via autocrine growth stimulation. PMID- 8402627 TI - A phase I clinical trial of murine monoclonal antibody D612 in patients with metastatic gastrointestinal cancer. AB - In a phase I study, 21 patients with metastatic adenocarcinoma of the gastrointestinal tract received the murine monoclonal antibody D612. This antibody is directed at a M(r) 48,000 antigen restrictively expressed on tumors of the gastrointestinal tract and to a limited degree on normal gastrointestinal mucosa. Patients received total doses of 10-180 mg/m2 administered as single or multiple doses of 1-100 mg/m2 over an 8-day period. Dose-limiting toxicity was secretory diarrhea. A single dose of 100 mg/m2 exceeded guidelines for maximal tolerated dose. Higher total doses were achieved in subsequent patients by using repeated administration of lower doses. Three of five patients receiving 60 mg/m2 for 3 doses (180 mg/m2 total dose) experienced grade 3 diarrhea and could not complete the prescribed course. The dose of 40 mg/m2 administered on days 1, 4, and 8 (total dose, 120 mg/m2) has been selected as the dose for phase II studies. The pharmacokinetics of D612 is best described by a one-compartment model with a mean t1/2 of 48 +/- 3 h (SEM). Eighteen of 21 patients developed human anti-mouse antibody (HAMA). Patients who developed high levels of HAMA demonstrated a more rapid clearance of the day 8 dose than those who developed low levels of HAMA. In all patients studied, a component of HAMA was directed at the D612 variable region. With one exception, serum from all patients with detectable antibody to the D612 variable region demonstrated anti-paratope reactivity. Thirty-four % of known metastatic sites demonstrated uptake of radiolabeled D612. There were no objective antitumor responses in this phase I trial. The antitumor effect of D612 in vitro has been shown to be potentiated by interleukin 2 and recombinant human macrophage colony-stimulating factor. A phase II study of D612 administered in combination with cytokines that enhance human effector function is presently ongoing. PMID- 8402628 TI - Inhibition of the in vivo conversion of androstenedione to estrone by the aromatase inhibitor vorozole in healthy postmenopausal women. AB - Vorozole is a new, potent, and highly selective nonsteroidal aromatase inhibitor, which in animal and human studies was found to be about 1000-fold more potent than aminoglutethimide. Almost all aromatase-inhibiting activity resides in the dextro-enantiomer currently undergoing clinical trials. A marked decrease in circulating estrogens was found in several studies of healthy premenopausal women and male volunteers treated with the racemate, referred to as vorozole racemate. To further evaluate the aromatase-inhibiting potency of this drug, the in vivo conversion of androstenedione to estrone was studied in 12 healthy postmenopausal women. Four h after a single oral dose of vorozole racemate, [14C]androstenedione and [3H]estrone were infused at a constant rate for 2 h. Women were randomized to receive vorozole racemate orally in one of three different doses, i.e., 1, 2.5, and 5 mg, in a double-blind protocol. Each woman acted as her own control in an identical experiment with a placebo carried out 2-4 weeks either before or after the test with vorozole racemate. In the urine, collected for 4 days after each experiment, estrogens were extracted and purified until a constant 3H/14C ratio of estrone was achieved. The percentage conversion of androstenedione to estrone in the 12 placebo experiments was 2.19 +/- 0.60% (mean +/- SD, n = 12). Following a single administration of vorozole racemate, the conversion decreased to 0.14 +/ 0.04%. The percentage inhibition was 93.0 +/- 2.5 (n = 4) following administration of 1 mg vorozole racemate; administration of 2.5 or 5 mg resulted in an inhibition percentage of 93.2 +/- 1.6 or 94.4 +/- 1.2, respectively. It is concluded that a single oral dose of 1-5 mg vorozole racemate results in an almost complete inhibition of in vivo aromatase activity. PMID- 8402629 TI - Modulation of cytotoxicity and cellular pharmacology of 1,2-diaminocyclohexane platinum (IV) complexes mediated by axial and equatorial ligands. AB - Isomers (R,R-, S,S-, and cis-) of 1,2-diaminocyclohexane (DACH) platinum(IV) complexes with selected axial and equatorial ligands were synthesized and evaluated for in vitro antitumor activity, cellular uptake, and total DNA-Pt adducts. L1210 cells, sensitive to cis-diamminedichloroplatinum(II) (CDDP) and tetraplatin (L1210/0), 160-fold resistant to CDDP [L1210/diamminedichloroplatinum (DDP)], or 70-fold resistant to tetraplatin (L1210/DACH), were used in conjunction with compounds having the general structure DACH-Pt(IV)-X2Y2, where X and Y are axial and equatorial ligands and X2Y2 are specifically Cl2Cl2,Ac2Cl2, (TFA)2Cl2, (OH)2Cl2, and Cl2CBDCA (Cl, chloro; Ac, acetato; TFA, trifluoroacetato; OH, hydroxo; CBDCA, 1,1-cyclobutanedicarboxylato). Comparison of cytotoxicities between isomers of Cl2Cl2,Ac2Cl2, or Cl2CBDCA indicated that R,R-isomers were the most effective against all three cell lines. The relatively lower activity of the S,S- and cis-isomers was cell line dependent: against L1210/DACH, both isomers of Cl2Cl2 were only 2- to 3-fold less effective, and this contrasted with 7- and 26-fold lower cytotoxicities, respectively, against L1210/DDP. Cross-resistance factors in the L1210/DDP and L1210/DACH lines depended on both isomeric form and the nature of axial or equatorial ligand. The L1210/DDP cells were 6- to 9-fold cross-resistant to the R,R-isomer, 8- to 15 fold to S,S-isomer, and 13- to 38-fold to cis-isomer. The L1210/DACH line was only 4- to 7-fold cross-resistant to the three isomers of Ac2Cl2 but cross resistance to the isomers was 47- to 79-fold for Cl2Cl2 and 22- to 56-fold for Cl2CBDCA complexes. Compared with CDDP, accumulation (2 h at 100 microM drug concentration) of Ac2Cl2 in the three L1210 cell lines was 26-50%, while uptake of Cl2Cl2 and (TFA)2Cl2 was 100-170% and 320-570%, respectively. The greatest DNA binding was seen with Cl2Cl2 in all cell lines, followed by (TFA)2Cl2, CDDP, and Ac2Cl2. DNA binding correlated directly with potency (1/concentration producing 50% inhibition) in the L1210/0 model (r = 0.973, P < 0.016) but not in the L1210/DDP and L1210/DACH models. Accumulation and DNA-binding studies indicated that binding efficiency to DNA was: Cl2Cl2 > Ac2Cl2 > CDDP > (TFA)2Cl2. In a nonreducing environment, the Pt(IV) complexes (20 microM) did not react with salmon sperm DNA. Reduced glutathione (100 microM), as a reducing agent, rendered full binding capacity to Cl2Cl2; binding was 25-30% of the expected maximum for (TFA)2Cl2, while Ac2Cl2 remained inert.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402630 TI - Estramustine depolymerizes microtubules by binding to tubulin. AB - To investigate the mechanism of action of the antineoplastic drug estramustine, we compared its effects on human prostate cancer cells with those of vinblastine. At their respective concentrations that result in 50% inhibition of clonogenic growth, both drugs caused an accumulation of cells blocked at mitosis and similar dose- and time-dependent depolymerization of interphase microtubules. Also, colcemid-resistant and colcemid-hypersensitive Chinese hamster ovary cells with tubulin mutations were collaterally cross-resistant or -sensitive to estramustine. Thus, the cytotoxicity of estramustine is due to its microtubule depolymerization properties. This could be caused by interaction with tubulin and/or with microtubule-associated proteins (MAPs). Previous investigations have shown that high concentrations of estramustine phosphate can inhibit microtubule polymerization in vitro by binding to MAPs. However, estramustine phosphate is the clinical prodrug to estramustine, the intracellular active compound. In this study, we investigated the effects of estramustine on the binding of MAPs to taxol-stabilized microtubules in vivo. In contrast to previous reports, no effect of estramustine on the binding of MAPs to microtubules was found. Furthermore, we found that polymerization of purified tubulin could be inhibited by estramustine in vitro. Taken together, these results demonstrate that estramustine causes depolymerization of microtubules by direct interaction with tubulin. PMID- 8402631 TI - Effect of a template-located 2',2'-difluorodeoxycytidine on the kinetics and fidelity of base insertion by Klenow (3'-->5'exonuclease-) fragment. AB - Gemcitabine [2',2'-difluorodeoxycytidine (dFdCyd)], a potent antitumor agent, inhibits DNA synthesis and is incorporated internally into DNA. The effect of a template-incorporated dFdCyd molecule (dFdCyd-) on DNA polymerase function was examined. Two 25-base deoxyoligonucleotides were synthesized with either a single dFdCyd- or template-incorporated deoxycytidine molecule (dCyd-) at the same position. Each was annealed separately to an identical complementary 5'-32P labeled primer and extended by the Klenow fragment (3'-->5' exo-) of DNA polymerase I. "Correct" insertion of dGMP was 80-fold less efficient opposite dFdCyd- than dCyd-. A comparison of misinsertion efficiencies opposite template dFdCyd gave values of 2.7 x 10(-2) for dAMP insertion, 1.1 x 10(-3) for dTMP insertion, and 5.9 x 10(-4) for dCMP insertion. A similar measurement opposite template dC gave values of 1.8 x 10(-4), 1.7 x 10(-4), and 2.9 x 10(-6) for dAMP, dTMP, and dCMP insertion, respectively. Thus, the presence of dFdCyd on the template strand inhibited "normal" DNA synthesis and increased deoxyribonucleotide misinsertion frequencies. Pausing during DNA synthesis occurred directly opposite template dFdCyd suggesting that dFdC.dG base pairs might be less stable than normal dC.dG pairs, resulting in a decreased rate of primer extension beyond this site. Consistent with kinetic data, thermal denaturation measurements using comparable surrounding sequences showed that dFdC.dG "correct" pairs were less stable than dC.dG base pairs. Measurements on base mispairs showed that dFdC.dC was more stable than dC.dC, while no measurable Tm differences were found between polymers containing dFdC.dA and dC.dA or dFdC.dT, and dC.dT. PMID- 8402632 TI - Polyethylene glycol-modified chimeric toxin composed of transforming growth factor alpha and Pseudomonas exotoxin. AB - Modification of proteins with monomethoxy-polyethylene glycol (mPEG) has been shown to prolong circulation time and to reduce immunogenicity. To make a mPEG modified recombinant toxin that retained cytotoxic activity but had a longer residence time in circulation, we have constructed an altered form of TGF alpha PE40, a recombinant toxin composed of human transforming growth factor alpha (TGF alpha) fused to a fragment of Pseudomonas exotoxin (PE38) devoid of its cell binding domain. In the newly designed protein, termed TGF alpha R29-L2-CH2-PE38QQ delta (TCP), there are no lysine residues in the TGF alpha and PE38 portions. Human IgG4 constant region CH2 and a tetradecapeptide linker; L2, are inserted between TGF alpha and PE38. Together, L2 and CH2 contain 13 lysine residues as potential modification sites for mPEG. mPEG conjugates of TCP (PEG-TCP) were generated and the products were resolved by ion exchange chromatography. Two PEG TCP species termed B4 and B6 retained 15 and 4% of cytotoxicity, respectively, and 26% of their receptor binding activity compared with the unmodified TCP. Both B4 and B6 had prolonged circulation times in the blood and reduced toxicity in animals. The mean residence times of B4 and B6 were 37 and 68 min, respectively, compared to 7 min for TCP. When administered i.v. to tumor bearing mice, both B4 and B6 produced marked antitumor effects whereas the unmodified TCP had none. Also, the immunogenicity of PEG-TCP was 5-10 times less than that of TCP. We suggest that the prolonged circulating time and reduced toxicity of PEG-TCP compensate for a diminished cytotoxic activity and enlarge significantly the therapeutic window of this chimeric toxin. PMID- 8402633 TI - In vitro and in vivo reversal of multidrug resistance by GF120918, an acridonecarboxamide derivative. AB - N-(4-[2-(1,2,3,4-tetrahydro-6,7-dimethoxy-2-isoquinolinyl)ethyl]- phenyl)-9,10 dihydro-5-methoxy-9-oxo-4-acridine carboxamide (GF120918) has been selected from a chemical program aimed at identifying an optimized inhibitor of multidrug resistance (MDR). The potency of GF120918 is assessed by dose-dependent sensitization of CHRC5, OV1/DXR and MCF7/ADR cells to the cytotoxicity of doxorubicin and vincristine respectively: GF120918 fully reverses multidrug resistance at 0.05 to 0.1 microM and is half maximally active at 0.02 microM. The spectrum of drugs sensitized by GF120918 coincides with those having the classical MDR phenotype. In CHRC5 cells, 0.01-0.1 microM GF120918 enhances the uptake of [3H]daunorubicin and blocks the efflux from preloaded cells. It is also shown that GF120918 is still active several hours after being taken away from the culture medium showing that it is not, like verapamil, effluxed rapidly by P glycoprotein. GF120918 effectively competes with [3H]azidopine for binding P glycoprotein, pointing to this transport membrane protein as its likely site of action. After i.v. administration to mice, GF120918 penetrates thoroughly various organs that have a tissue level/blood level ratio above 10. It is eliminated from organs and blood with a half-time of approximately 2.7 h. It is well absorbed after p.o. administration. In mice implanted i.p. with the MDR P388/Dox tumor, a single i.v. or p.o. dose of GF120918 restores sensitivity of the tumor to a single i.p. dose (5 mg/kg) of doxorubicin administered 1 h later. A statistically significant effect is observed at 1 mg/kg GF120918 i.v. and maximal effect is reached at 5 mg/kg. Similarly, whereas neither drug alone is effective, GF120918 (10 mg/kg i.p.) associated with doxorubicin (5 mg/kg i.p.) inhibits the growth of the moderately MDR C26 tumor implanted s.c. as assessed by tumor size at day 19. GF120918 does not modify significantly the distribution or the elimination of doxorubicin in mice ruling out the possibility that the antitumor effects seen might be explained by pharmacokinetic interactions. PMID- 8402634 TI - Cardioprotective properties of O-(beta-hydroxyethyl)-rutosides in doxorubicin pretreated BALB/c mice. AB - Chronic doxorubicin-induced cardiotoxicity is believed to be caused by the formation of oxygen free radicals. Thus O-(beta-hydroxyethyl)-rutosides, a standardized flavonoid mixture (Venoruton) with iron chelating and radical scavenging activity, might provide protection. Therefore, we investigated the (cardio)protective effect of Venoruton (1.5 g/kg injected i.p. on days 1-5, 8-12, 15-19, and 22-26) in BALB/c mice treated with doxorubicin (4 mg/kg injected i.v. on days 1, 8, 15, and 22) compared with mice treated with doxorubicin alone. Saline-treated animals served as controls. No mortality was encountered in either of the groups; weight gain data suggest little general toxicity of this dose schedule. The basal frequency of the isolated right atria was increased in doxorubicin-pretreated animals as compared to control animals (468 +/- 22 and 366 +/- 20 beats/min, respectively). Venoruton coadministration diminished this increase (373 +/- 17 beats/min). The -log of the concentration giving 50% effect of l-isoprenaline on the right atrium was changed after doxorubicin pretreatment (8.33 +/- 0.04 versus 8.86 +/- 0.06 for control animals). Venoruton coadministration resulted in a smaller shift in the -log of the concentration giving 50% effect (8.51 +/- 0.10) than with doxorubicin alone. The extent of cardiotoxicity found in the functional studies was confirmed by histological scoring of heart ventricle damage. It can be concluded that Venoruton has the potential to protect against doxorubicin-induced cardiotoxicity. PMID- 8402635 TI - Main drug- and carcinogen-metabolizing enzyme systems in human non-small cell lung cancer and peritumoral tissues. AB - To better understand the importance of drug-metabolizing enzymes in carcinogenesis and anticancer drug sensitivity of human non-small cell lung cancer, we studied the main drug-metabolizing enzyme systems in both lung tumors and their corresponding nontumoral lung tissues in 12 patients. The following enzymes were assayed by Western blot analysis: cytochromes P-450 (1A1/A2, 2B1/B2, 2C8-10, 2E1, 3A4); epoxide hydrolase; and glutathione S-transferase isoenzymes (GST-alpha, -mu, and -pi). The activity of the following enzymes or cofactor were determined by spectrophotometric or fluorometric assays: glutathione S transferase (GST); total glutathione; UDP-glucuronosyltransferase; beta glucuronidase; sulfotransferase; and sulfatase. Results showed the presence of cytochrome P-450 1A1/1A2 in both tumoral and nontumoral tissues. P-450 1A1/1A2 levels were 3-fold lower in tumors compared to corresponding nontumoral tissues (P < 0.05). None of the other probed cytochromes P-450 were detected in either tumoral or nontumoral lung tissues. For the glutathione system, no significant difference between tumoral and nontumoral tissues was observed (GST activity, glutathione content, GST-alpha, -mu, and -pi). A positive linear correlation was observed between GST activity and GST-alpha or GST-pi. No significant difference was observed for the glucuronide and the sulfate pathways and their corresponding hydrolytic enzymes. Epoxide hydrolase was significantly decreased in tumors compared to nontumoral lung tissues (P < 0.05). In conclusion, these results showed differences between non-small cell lung tumors and nontumoral tissues for cytochrome P-450 1A1/1A2 and epoxide hydrolase. These differences between tumors and peritumoral tissues with regard to these drug-metabolizing enzymes could reflect differences occurring after malignant transformation and may play a role in drug sensitivity to anticancer drugs. PMID- 8402636 TI - Caffeine prevents apoptosis and cell cycle effects induced by camptothecin or topotecan in HL-60 cells. AB - Caffeine (3,7-dihydro-1,3,7,-trimethyl-1H-purine-6,6-dione; CAF) is known to potentiate the cytotoxic effects of DNA damaging agents such as ionizing radiation and alkylating agents. In contrast, however, the cytotoxic and cytostatic activity of aromatic, DNA-intercalating, DNA topoisomerase II inhibitors such as Adriamycin, ellipticine, or mitoxantrone are diminished in the presence of CAF. To resolve whether the protective effect of CAF is associated with a particular mechanism of drug interaction (e.g., intercalation into DNA, inhibition of DNA topoisomerase II), or the aromatic nature of the drug structure, per se, we have presently studied the effects of CAF on the cytostatic and cytotoxic action of camptothecin (CAM) and its less toxic but more water soluble derivative topotecan (TPT) on HL-60 human myelogenous leukemia cells: both drugs have aromatic structures but are nonintercalating inhibitors of DNA topoisomerase I. By using spectroscopy and titration microcalorimetry, we have also studied the direct interaction between CAF and TPT in solution. Low (20 nM) concentrations of CAM or TPT perturbed progression of HL-60 cells through S phase, whereas higher concentrations (0.15 microM) of these drugs induced apoptosis; both effects were easily demonstrable after 4 h of treatment. When added simultaneously with CAM or TPT, CAF prevented both effects. The protective effect of CAF was concentration dependent and evident within the concentration range of 1-5 mM; nearly total protection was seen at a CAF concentration of 5 mM. The bathochromic and hypochromic shift in the absorption spectrum of the water soluble compound TPT upon addition of CAF indicated that CAF and TPT interact (stack) in a fashion similar to that previously observed for CAF and DNA intercalators. Microcalorimetric measurements of TPT titration with CAF indicate an exothermic reaction between these compounds (the enthalpy change was delta H degree = -4.2 kcal/mol), which is consistent with a stacking model of CAF-TPT interaction. Thus, the ability of CAF to protect HL-60 cells against the cell kinetic effects of CAM or TPT, as in the case of DNA intercalating topoisomerase II inhibitors, is most likely due to formation of complexes between CAF and these aromatic molecules, which result in reducing the effective concentration of the free form of these drugs available to the cells. PMID- 8402637 TI - In vivo antitumor activity of 5-fluorocytosine on human colorectal carcinoma cells genetically modified to express cytosine deaminase. AB - A human colorectal carcinoma cell line, WiDr, was genetically engineered to express the nonmammalian enzyme, cytosine deaminase (CD). Expression of CD in WiDr cells (WiDr/CD) did not alter the growth rate of these cells when grown in vitro or as solid tumor xenografts in nude mice. However, expression of CD did increase the sensitivity of these cells to the nontoxic prodrug, 5-fluorocytosine (FCyt), decreasing the 50% inhibitory concentration for FCyt from 26,000 microM in parental WiDr cells to 27 microM in WiDr/CD cells. The increase in sensitivity to FCyt in WiDr/CD cells was the result of the CD-mediated conversion of FCyt to 5-fluorouracil (FUra) and subsequent FUra anabolites. The half-life of the prodrug, FCyt, was determined to be approximately 40 min in nude mice. A single i.p. injection of 500 mg FCyt/kg body weight resulted in a transient FCyt plasma level of approximately 4000 microM while osmotic minipumps or constant tail vein infusions of FCyt achieved continual FCyt plasma levels of 5 microM and 50 microM, respectively, with no overt signs of toxicity. Significant antitumor effects were observed in nude mice bearing tumors derived from WiDr/CD cells when these animals were given 500 mg FCyt/kg i.p. for 10 consecutive days. These antitumor effects were demonstrated by decreases in tumor growth rate, tumor size, tumor weight, and thymidine incorporation into tumor DNA. This antitumor effect was significant but less profound if FCyt was administered by constant tail vein infusion. WiDr and WiDr/CD cells were very sensitive to FUra in vitro (50% inhibitory concentration approximately 5 microM). However, no significant antitumor effects were observed in nude mice bearing tumors derived from either WiDr or WiDr/CD cells when these animals were treated with various doses of FUra. Taken collectively, these data indicate that nontoxic plasma levels of FCyt can be attained which can produce profound antitumor effects on tumors engineered to express CD and that these antitumor effects are significantly better than those that can be achieved using FUra. These positive data support the continued development of a gene therapy approach to colorectal carcinoma involving the selective expression of CD in colorectal tumors with subsequent administration of FCyt. PMID- 8402638 TI - Reversible inhibition of proliferation of epithelial cell lines by Agaricus bisporus (edible mushroom) lectin. AB - Galactosyl beta-1,3-N-acetyl galactosamine (Gal beta-1,3-GalNAc) (Thomsen Friedenreich antigen), the Class I core sequence in O-linked oligosaccharide chains, behaves as an oncofetal antigen showing increased expression in many epithelial malignancies. Previous work has shown that peanut agglutinin (PNA), a lectin that binds Gal beta-1,3-GalNAc, stimulates proliferation in HT-29 (human colon cancer) cells and normal human colonic epithelium and this implies that cell surface glycoproteins which express Gal beta-1,3-GalNAc may play an important role in the regulation of epithelial cell proliferation. We have now studied the effect on epithelial cells of another dietary Gal beta-1,3-GalNAc binding lectin, the edible mushroom Agaricus bisporus lectin (ABL). This differs from PNA in its ability to bind also to sialylated Gal beta-1,3-GalNAc. In contrast to PNA, ABL (25 micrograms/ml) inhibited incorporation of [3H]-thymidine into DNA of HT29 colon cancer cells by 87% (95% confidence limit, 85-89%), Caco-2 colon cancer cells by 16% (95% confidence limit, 12-20%), MCF-7 breast cancer cells by 50% (95% confidence limit, 47-52%), and Rama-27 rat mammary fibroblasts by 55% (95% confidence limit, 51-60%) when these cells were grown for 24 h in serum-free medium. When assessed by cell count, similar inhibition of proliferation of HT29 cells by ABL was found. In the presence of 2% fetal calf serum (which contains the ABL-binding glycoprotein fetuin), the inhibitory effect of ABL on cell proliferation was still demonstrable but at increased ABL concentration (60 micrograms/ml for 49% inhibition). Ten micrograms/ml ABL completely abolished the stimulatory effect on [3H]thymidine incorporation of epidermal growth factor (100 pg/ml) and PNA (25 micrograms/ml) and markedly inhibited the stimulatory effect of insulin (50 ng/ml). ABL (0.2 mg/ml) caused no cytotoxicity to HT29, MCF-7, and Rama-27 cells as measured by trypan blue exclusion, and inhibition of proliferation in HT29 cells caused by 50 micrograms/ml ABL was reversible after removal of the lectin. Binding studies with 125I-labeled ABL suggested a single class of binding site with an apparent Kd value of (4.12 +/- 0.29) x 10(-7) M with (3.6 +/- 0.3) x 10(7) binding sites/cell. A. bisporus lectin is a reversible noncytotoxic inhibitor of epithelial cell proliferation which deserves study as a potential agent for cancer therapy. PMID- 8402639 TI - Tumor-specific, schedule-dependent interaction between tirapazamine (SR 4233) and cisplatin. AB - Tirapazamine (SR 4233), a benzotriazine di-N-oxide, a potent and selective killer of hypoxic cells, is currently in Phase I clinical trials with the expectation that it will be combined with radiation therapy. However, because of the likelihood that hypoxic tumor cells may also be resistant to some commonly used chemotherapeutic agents, we have tested tirapazamine in combination with cisplatin (c-DDP) in the mouse RIF-1 tumor. A large, schedule-dependent enhancement of tumor cell killing was observed both in vivo and in vitro, with a maximal response observed when the SR 4233 was given 2-3 h before c-DDP. Assay of serum blood urea nitrogen levels following treatment with these two drugs indicates that SR 4233 does not enhance the kidney damage which can result from high doses of c-DDP. Leukopenia induced by the two drugs in combination was equal to that predicted from an additive effect of the responses to the individual drugs. Also, there was no change in the systemic toxicity of c-DDP (as judged by 50% lethal dose) when SR 4233 was combined with c-DDP at a dose and timing that produced the maximum tumor interaction. These observations point to a promising new combination therapy with considerable therapeutic advantage. PMID- 8402640 TI - Antitumor effect of anti-epidermal growth factor receptor monoclonal antibodies plus cis-diamminedichloroplatinum on well established A431 cell xenografts. AB - We have explored the therapeutic effects of anti-epidermal growth factor receptor monoclonal antibodies (MAbs) 225 and 528 on well established A431 epidermoid carcinoma xenografts, approximately 400 mm3 (1 cm in diameter) at the start of treatment. In previous reports we demonstrated that MAbs 225 and 528 prevented the growth of A431 cell xenografts in nude mice when treatment was begun on the day of tumor cell inoculation. Since anti-epidermal growth factor receptor MAb therapy of well established tumors was unable to retard growth, we explored combination therapy with MAb plus the chemotherapeutic agent cis diamminedichloroplatinum (cis-DDP). Additive and concentration-dependent growth inhibitory effects of MAb with cis-DDP were observed in cultures of A431 cells. Neither intensive treatment with 225 MAb (1 mg/mouse, i.p. on day 8 after tumor inoculation, and twice weekly for 4 weeks) nor a maximally tolerated single dose of cis-DDP [150 micrograms/25 g (6 mg/kg) mouse weight, i.p. on day 8] had significant effects on tumor growth. However, the two treatments in combination resulted in substantial xenograft growth inhibition, compared with both an untreated control group and animals treated with a single modality. When a second dose of cis-DDP (150 micrograms/25 g) was added after 10 days, combination therapy with 225 MAb produced striking antitumor effects. At the end of 1 month tumor xenografts had disappeared in all but one mouse, and no tumor relapses occurred during 6 months of observation. Identical results were obtained with anti-epidermal growth factor receptor MAb 528 in combination with cis-DDP. The results of these studies provide a novel approach to the treatment of well established tumor xenografts, which may have application in the therapy of human malignancies. PMID- 8402641 TI - Inhibitory effect of interleukin 4 on production of interleukin 6 by adult T-cell leukemia cells. AB - Freshly isolated leukemic cells from patients with adult T-cell leukemia (ATL) produce high levels of interleukin 6 (IL-6), which is suggested to play an important role in thrombocytosis, elevation of C-reactive protein, and hypercalcemia in ATL. In this study, we investigated the effects of T-cell growth factors such as interleukin 2 (IL-2) and interleukin 4 (IL-4) on IL-6 production by ATL cells in vitro. Although IL-2 and/or IL-4 enhanced the cell proliferation of freshly isolated ATL cells from seven of nine patients, IL-2 did not affect the IL-6 release in most cases. In contrast, another T-cell tropic factor, IL-4 markedly inhibited the release of IL-6 in the conditioned medium in all cases. This IL-4-mediated inhibition of IL-6 release was completely abrogated by the addition of anti-IL-4 monoclonal antibody. Time course experiments demonstrated that IL-4 reduced the secretion of IL-6 for a prolonged period of time (more than 72 h). By Northern analysis, IL-4 reduced the transcription level of IL-6 mRNA. Furthermore, by flow cytofluorometry with the use of anti-human IL-4 receptor monoclonal antibody, ATL cells showed the significant level of IL-4 receptor on their cell surfaces without any stimulation. These data suggest that IL-4 may play an important regulatory role in the production of IL-6 in ATL. PMID- 8402642 TI - Immune reactions against hepatitis B viral antigens lead to the rejection of hepatocellular carcinoma in BALB/c mice. AB - Human hepatitis B virus (HBV) is closely associated with hepatocellular carcinoma. However, the mechanism of carcinogenesis and the immune responses to HBV infection and hepatocellular carcinoma are not clearly understood. Recently, we established BALB/c mouse liver (ML) cell lines and demonstrated that transfection of ML cell lines with HBV dimer DNA resulted in the expression of HBV antigens (1). The HBV-transfected ML cells and the parental ML cells showed similar tumorigenicity in nude mice. However, the HBV-transfected cells had much lower tumorigenicity in BALB/c mice. Similar results were also obtained in two cloned ML cell lines, ML-1.1 and ML-1.2, transfected with plasmid DNA containing HBs, HBc, or HBx gene. Furthermore, adoptive transfer of spleen cells from BALB/c mice immunized with HBsAg- or HBcAg-expressing ML-1.1 cells caused regression of tumor cells expressing the corresponding antigens in nude mice. In addition, transfer of spleen cells from BALB/c mice immunized with purified HBsAg or HBcAg also caused tumor regression. These results demonstrate that HBsAg and HBcAg can induce immunity which leads to the rejection of hepatocellular carcinoma in vivo. PMID- 8402643 TI - Effect of verapamil on doxorubicin cardiotoxicity: altered muscle gene expression in cultured neonatal rat cardiomyocytes. AB - Verapamil reverses multidrug resistance acquired by cancer cells during treatment with chemotherapeutic agents such as doxorubicin by inhibiting the function of P glycoprotein. Verapamil has also been suggested to potentiate the cardiotoxicity of doxorubicin. We have recently demonstrated that selective inhibition of cardiac muscle gene expression is among the earliest events in doxorubicin cardiotoxicity. To explore the influence of verapamil on doxorubicin cardiotoxicity, we evaluated [14C]-doxorubicin accumulation, cardiac muscle gene expression by Northern blot analysis, and ultrastructural changes in cultured cardiomyocytes in the presence and absence of verapamil. Treatment with a combination of doxorubicin and verapamil for 24 h did not augment doxorubicin accumulation in cardiomyocytes, although substantial augmentation of doxorubicin accumulation by verapamil in cardiac fibroblasts was observed. Further, treatment with verapamil for 24 h did not augment the decrease in expression of muscle genes induced by doxorubicin (myosin light chain 2 slow, troponin I, M isoform creatine kinase). However, we found that verapamil reduced alpha-actin gene expression in a direct, doxorubicin-independent manner. Furthermore, the effect of doxorubicin plus verapamil on alpha-actin gene expression was additive over a wide range of doxorubicin and verapamil concentrations, resulting in a selective augmentation of doxorubicin-induced inhibition of gene expression for this single muscle protein gene. This was reflected in a substantial increase in cardiac myocyte damage when treatment with verapamil and doxorubicin was compared to treatment with doxorubicin alone by thin section electron microscopy. This suggests a possible mechanism by which verapamil may potentiate doxorubicin cardiotoxicity. PMID- 8402644 TI - p53 mutations and histological type of invasive breast carcinoma. AB - A polymerase chain reaction-single strand conformation polymorphism assay was used to assess p53 mutations in 148 invasive breast carcinomas, selected on the basis of their histotype. They comprised 56 lobular, 47 ductal, 19 mucinous, 18 medullary, and 8 papillary carcinomas. The distribution of p53 mutations was significantly different (P = 0.006) in the histotypes examined: mutations were frequent in medullary (39%) and ductal (26%), less common in lobular (12%), and absent in mucinous and papillary carcinomas. The frequency of mutations in the exon 5 of the p53 gene was significantly higher in medullary carcinomas than in the other histotypes: 5 (63%) of the mutations found in exon 5 were observed in medullary carcinomas (P = 0.012). One hundred twenty-two tumors from the total were also examined by immunohistochemistry for p53 overexpression using antibody PAb 1801. A specific immunostaining in neoplastic cells was present in 12 tumors. A strong correlation (P < 0.001) was observed between p53 mutations and nuclear accumulation of the p53 protein: 10 tumors were scored positive for both p53 mutation and overexpression. However, in 9 cases having a mutated p53 gene we failed to find a positive immunoreaction. A significant association (P = 0.01) was present between mutations in the p53 gene and high proliferative activity of the tumors determined by immunohistochemistry with monoclonal antibody Ki-67. Moreover, a significantly higher expression of the Ki-67 antigen was found in medullary carcinomas compared to the other histotypes. Our findings indicate that in invasive breast carcinomas structural abnormalities of the p53 gene are mainly seen in medullary and ductal tumors and that the other histological types, especially those associated with a high level of differentiation and favorable prognosis, show a very low incidence of p53 mutations. PMID- 8402645 TI - Clonal analysis by study of X chromosome inactivation in formalin-fixed paraffin embedded tissue. AB - Analysis of clonality by X chromosome inactivation has proven to be a powerful strategy in the study of neoplastic and preneoplastic disorders (P. J. Fialkow, Biochim. Biophys. Acta, 458: 283-321, 1976; B. Vogelstein et al., Cancer Res., 47: 4806-4813, 1987). Recently, the gene for the androgen receptor has been shown to be a highly polymorphic locus in which methylation of DNA correlates with inactivation of one or the other X homologue (R. C. Allen et al., Am. J. Hum. Genet., 51: 1229-1239, 1992). Unlike other loci used for analysis of X inactivation, the methylation sites within the androgen receptor gene lie close to the region of DNA containing the polymorphism. Consequently, it should be possible to use methylation-sensitive restriction enzymes and polymerase chain reaction to study differential methylation among alleles of this gene in formalin fixed and paraffin-embedded archival tissue specimens. To investigate this question, we performed clonal analysis on a variety of randomly selected, formalin-fixed, paraffin-embedded tumor tissues from 15 women. Thirteen of the women were found to be heterozygous for polymorphisms at the androgen receptor locus. Among these women, 11 tumors were clearly clonal in assays of methylation of the androgen receptor gene. Interpretation of results for the remaining two cases was complicated because of a skewed pattern of X chromosome inactivation found in normal control tissues. We conclude that analysis of methylation in the androgen receptor gene should allow study of clonality in most formalin-fixed, paraffin-embedded tissue specimens from women, including small preneoplastic lesions or rare conditions for which sufficient material is not available for analysis by other techniques. PMID- 8402646 TI - Effect of diets containing different levels of linoleic acid on human breast cancer growth and lung metastasis in nude mice. AB - The purpose of the study was to determine the effect of three different levels of dietary linoleic acid (LA) intake on the growth of MDA-MB-435 human breast cancer cells in the mammary fat pads of nude mice, and their metastasis to the lungs. These diets were isocaloric, and contained different mixtures of safflower (LA rich) and coconut (saturated fatty acid-rich) oils to provide 23% (w/w) total fat, with 2, 8, and 12% (w/w) LA. A fourth group was fed a low-fat, 5% (w/w) corn oil diet. There were 25 mice in each dietary group. A necropsy, 12 weeks after the tumor cell injections, the primary tumor weights in the 12% LA (4.1 +/- 2.7 g)- and 8% LA (3.5 +/- 1.7 g)-fed groups were significantly greater (P < 0.05) than those fed the 2% LA diet (2.5 +/- 1.5 g); they did not differ significantly from the weights of mammary fat pad tumors in the 5% corn oil-fed mice. The incidence of grossly visible pulmonary metastatic nodules was not significantly different between the 8 and 12% LA-fed mice, but was higher for both groups compared with the 2% LA-fed group (P < 0.05), with a similar trend in comparison with the 5% corn oil group. The mean total calculated volumes of the macroscopic metastases per tumor-bearing mouse were significantly greater in the 8 and 12% LA (157 +/- 250.7 and 99.1 +/- 140.0 mm3, respectively), compared with the 2% LA (23.3 +/- 51.8 mm3)- and 5% corn oil (24.5 +/- 35.1 mm3)-fed mice; all P < 0.05. Micrometastases were observed most frequently in the 5% corn oil and 2% LA dietary groups, but none of the differences were statistically significant. No differences were detected in the concentrations of prostaglandin E, leukotriene B4, or 5-hydroxyeicosatetraenoic acid in tumors from mice fed the four different diets. PMID- 8402647 TI - Expression of p53 gene in 184 unifocal hepatocellular carcinomas: association with tumor growth and invasiveness. AB - To elucidate the biological significance of the p53 gene expression in human hepatocellular carcinoma (HCC), the p53 protein was studied in 184 resected unifocal primary HCCs, including 102 small (< or = 5 cm) and 82 large HCCs (> 5 cm), using immunocytochemistry. The p53 mRNA expression was analyzed in 69 cases using Northern hybridization. The p53 protein, which was detected in 58 HCCs (31.5%), was overexpressed more frequently in HCC with elevated serum alpha fetoprotein level (37.9 versus 25%, P < 0.04), in large HCC (39.0 versus 25.5%, P < 0.03), and in invasive HCC (35.1 versus 13.3%, P < 0.01). The overexpression of p53 protein closely correlated with p53 mRNA overexpression (75 versus 44.4%, P < 0.003), and p53 gene mutation (76.9 versus 19.2%, P < 1 x 10(-9)). HCCs with p53 protein expression (group A) and those negative for both p53 protein and mRNA expression (group B) had an unfavorable outcome, while HCC with no p53 protein but with p53 mRNA overexpression (group C) had the best outcome; the 4-year survival was 26.1, 26.3, and 62.5%, respectively. The p53 gene mutation was significantly higher in group A HCC (76.9%) than in groups B (27.3%) and C (23.5%), P < 0.0001. The results suggest that the p53 protein and mRNA expression patterns in HCC correlate with p53 gene mutation and tumor behavior and may serve as a molecular prognostic factor. PMID- 8402648 TI - Investigation of the subcellular distribution of the bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes. AB - A multidisciplinary approach was taken to investigate the intracellular locations of the 26-kDa integral membrane protein encoded by the bcl-2 gene. Subcellular fractionation analysis of a t(14;18)-containing lymphoma cell line revealed the presence of Bcl-2 protein in nuclear, heavy-membrane, and light-membrane fractions but not in cytosol. Sedimentation of heavy-membrane fractions in Nycodenz and Percoll continuous gradients demonstrated comigration of p26-Bcl-2 with mitochondrial but not other organelle-associated proteins. Fractionation of light-membrane fractions using discontinuous sucrose-gradients revealed association of Bcl-2 protein primarily with lighter-density microsomes (smooth endoplasmic reticulum) as opposed to heavy-density microsomes (rough endoplasmic reticulum). Immune microscopy studies using laser-scanning microscopy, pre- and postembedding electron microscopic methods, and six different anti-Bcl-2 antibodies demonstrated Bcl-2 immunoreactivity in the nuclear envelope and outer mitochondrial membrane in a patchy distribution. Furthermore, anti-Bcl-2 antibody immunoreactivity generally appeared to directly overlie the nuclear envelope in high magnification electron microscopic studies, reminiscent of nuclear pore complexes. Addition of in vitro translated p26-Bcl-2 to isolated translocation competent mitochondria revealed transmembrane domain-dependent association of Bcl 2 protein with mitochondria but provided no evidence for import into a protease resistant compartment, consistent with immunomicroscopic localization to the outer mitochondrial membrane. Taken together, the findings demonstrate that p26 Bcl-2 resides primarily in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membrane in a nonuniform distribution suggestive of participation in protein complexes perhaps involved in some aspect of transport. PMID- 8402649 TI - Immunodetection of endogenous opioid peptides in human brain tumors and associated cyst fluids. AB - The antitumorigenic effects of endogenous opioid peptides and their presence in extracerebral tumors are well documented. In this study, methionine-enkephaline (met-enkephalin) was measured by radioimmunoassay in 108 glial and nonglial brain tumors and in 44 associated cyst fluids. By immunohistochemistry, the distribution of the peptide and its precursor, preproenkephalin A, was also analyzed. Met-enkephalin and preproenkephalin were detected in the cytoplasm and cell processes of all tumors. Moreover, for neuroectodermal tumors (i.e., gliomas, gangliogliomas, and dysembryoplastic neuroepithelial tumors), a strong inverse correlation (P < 0.0001) was observed between the met-enkephalin levels and the degree of malignancy (242.9, 148.3, 55.3, and 30.3 pg/mg protein for grade 1, 2, 3, and 4, respectively). When compared to normal tissue, this differential expression mainly results from a decrease in the opioid peptide content in high-grade neuroectodermal tumors. Meningiomas and cerebral metastases displayed low met-enkephalin levels, similar to those of grade 4 neuroectodermal tumors. Large amounts of met-enkephalin were found in all cyst fluids. These data suggest that the endogenous opioid system is an integral component of brain tumors and that met-enkephalin may represent a useful malignancy marker in neuroectodermal tumors. PMID- 8402650 TI - Expression of vascular permeability factor (vascular endothelial growth factor) and its receptors in adenocarcinomas of the gastrointestinal tract. AB - Vascular permeability factor (VPF) is one of the most potent known inducers of microvascular hyperpermeability; in addition, it is a selective endothelial cell growth factor, hence its alternate name, vascular endothelial growth factor. VPF exerts its actions on the microvasculature by interacting with specific endothelial cell receptors. VPF is expressed by many transplantable animal tumors, by tumor cell lines in culture, and by certain normal cells in situ. The purpose of the present investigation was to determine whether and with what consistency VPF and its endothelial cell receptors are expressed in primary autochthonous human tumor of gastrointestinal tract origin, as determined by in situ hybridization and immunohistochemistry. Twenty-one primary adenocarcinomas (17 colon, 2 stomach, 1 small bowel, and 1 pancreas) were studied. The malignant epithelial cells expressed VPF mRNA strongly, in contrast to normal epithelium, hyperplastic polyps, and adenomas, which expressed little or no VPF mRNA. VPF expression was further increased in tumor cells immediately adjacent to zones of tumor necrosis; in such areas, occasional stromal cells also expressed VPF mRNA. In the ten colon carcinomas studied, tumor cells stained for VPF protein by immunohistochemistry. The endothelial cells of nearby stromal blood vessels also stained for VPF by immunohistochemistry and in addition expressed mRNAs encoding the VPF receptors flt-1 and kdr as determined by in situ hybridization. Endothelial cells away from the tumor did not stain for VPF and no definite mRNA expression for flt-1 or kdr was detected by in situ hybridization. The ganglion cells of the myenteric plexus of normal bowel expressed VPF mRNA and protein. These data indicate that primary autochthonous human tumors of gastrointestinal origin regularly express both VPF mRNA and VPF protein and that adjacent stromal vessels express mRNAs for both known VPF receptors. VPF is likely to contribute to tumor growth by promoting angiogenesis and stroma formation, both directly, through its action as an endothelial cell growth factor, and indirectly, by increasing vascular permeability, thereby leading to plasma protein extravasation, fibrin deposition, and the eventual replacement of the resulting matrix with vascularized stroma. PMID- 8402651 TI - Aberrant expression of type I fibroblast growth factor receptor in human pancreatic adenocarcinomas. AB - Acidic and basic fibroblast growth factors are mitogenic polypeptides that are overexpressed in pancreatic cancer. To determine whether fibroblast growth factors may exert direct effects on pancreatic cancer cells in vivo, we compared the expression of the high-affinity type I fibroblast growth factor receptor (FGFR-1) in human pancreatic tissues. In the normal pancreas, FGFR-1 immunostaining was seen mainly in acinar cells. In pancreatic cancers, FGFR-1 was abundant in ductal-like cancer cells which also exhibited many FGFR-1 mRNA in situ hybridization grains. Analysis by the polymerase chain reaction and RNase protection revealed that the 2-immunoglobulin-like and the 3-immunoglobulin-like forms of FGFR-1 were expressed in all tissue samples, and that the 2 immunoglobulin-like form was overexpressed in the cancer tissues by comparison with the normal tissues. These findings suggest that the 2-immunoglobulin-like form of FGFR-1 may contribute to aberrant autocrine and paracrine pathways in pancreatic cancer. PMID- 8402652 TI - Evidence for oligoclonal T-cell response in a metastasis of renal cell carcinoma responding to vaccination with autologous tumor cells and transfer of in vitro sensitized vaccine-draining lymph node lymphocytes. AB - Peripheral blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TIL) of a patient with von Hippel-Lindau disease and renal cell carcinoma were studied for the T-cell receptor beta chain variable region (TCR-V beta) repertoire. The patient was vaccinated with irradiated autologous tumor cells from a renal tumor mass, a vaccine-draining lymph node was removed, and lymphocytes were cultured in the presence of autologous tumor cells and low-dose interleukin 2 (IL2). These lymphocytes were adoptively transferred to the patient together with systemic IL2 (30,000 IU/kg every 8 h). Analysis of TCR-V beta expression was performed by polymerase chain reaction in PBL before, during, and after therapy, in vaccine draining lymph node lymphocytes, and in TIL obtained from moderately infiltrated, nonresponding renal tumor mass and from a more intensely infiltrated lung metastasis, which was responding to treatment. Significant differences in the expression of TCR-V beta 13.1 by T-cells recovered from these various sites were observed. Also, TIL recovered from the responding lung metastasis and cultured in the presence of IL2 gave rise to autologous tumor-reactive CD4+ T-cells, whereas the nonresponsive renal tumor yielded a mixture of T- and natural killer cells. In PBL obtained prior to treatment and during IL2 therapy, expression of V beta 13.1 was 0.7 and 1.8%, respectively, of the total V beta gene repertoire. Fresh vaccine-draining lymph node lymphocytes contained 5.9% of V beta 13.1-expressing T-cells. After IL2 therapy, V beta 13.1 gene expression increased to 5.4% in PBL. In the nonresponding tumor mass, the frequency of V beta 13.1 gene expression among TIL was 12%, whereas in the responding, highly infiltrated nodule, it was 28%, with a striking loss of expression of other V beta gene families. Sequencing of the amplified product of V beta 13.1 complementary DNA from the responding pulmonary metastasis showed restrictions in the complementarity-determining region 3. Thus, in vivo expansion of V beta 13.1-expressing CD4+ T-cells, possibly in response to a tumor-associated antigen, occurred in the responding tumor mass following this form of therapy and correlated with tumor course. PMID- 8402653 TI - A single amino acid change in human O6-alkylguanine-DNA alkyltransferase decreasing sensitivity to inactivation by O6-benzylguanine. AB - Mammalian O6-alkylguanine-DNA alkyltransferases (AGTs) are readily inactivated by incubation with the pseudosubstrate, O6-benzylguanine, but the equivalent protein from the Escherichia coli ogt gene is much less sensitive and the Saccharomyces cerevisiae and E. coli ada gene product AGTs are completely resistant to this compound. We have expressed the normal human AGT and various point mutations (C145A, W100A, and P140A) in an ada- ogt- strain of E. coli and tested these proteins against DNA substrates containing O6-methylguanine, for inactivation by O6-benzylguanine and for the ability to produce guanine from O6-benzylguanine. The C145A mutation was inactive as expected since this residue forms the methyl acceptor site. Mutants W100A and P140A were fully active against methylated DNA substrates but the P140A mutant was much less sensitive to inactivation by O6 benzylguanine and failed to form significant amounts of [3H]guanine when incubated with O6-benzyl[8-3H]-guanine. The proline at position 140 in mammalian AGTs is replaced by alanine in the Ada and yeast AGTs and by serine in the Ogt AGT. These results suggest that this proline residue affects the configuration of the active site allowing the O6-benzylguanine to enter and react with the mammalian AGT. The production of resistance to O6-benzylguanine by a single base change raises the possibility that such resistance may arise quite readily in cells of tumors treated therapeutically with the combination of O6-benzylguanine and an alkylating agent. PMID- 8402654 TI - Altered trans-activational properties of a mutated WT1 gene product in a WAGR associated Wilms' tumor. AB - WAGR syndrome is an acronym for a rare constellation of congenital abnormalities including predisposition to Wilms' tumor, Aniridia, Genitourinary malformations, and mental Retardation. These congenital defects are associated with a constitutional deletion affecting one copy of chromosome band 11p13, implicating the loss of one allele from a number of contiguous genes in this syndrome. Predisposition to Wilms' tumor and genitourinary abnormalities have been attributed to hemizygosity for the WT1 tumor suppressor gene, a transcriptional repressor that is normally expressed transiently during kidney development. Here we show that a Wilms' tumor arising in a child with WAGR syndrome contained a point mutation within the remaining WT1 allele. This mutation resulted in a glycine to aspartic acid substitution within the putative trans-activation domain of WT1, converting the encoded protein from a transcriptional repressor to an activator of its target DNA sequence. Thus, a critical amino acid substitution can alter the functional properties of WT1 and provide the "second hit" required for Wilms tumorigenesis. PMID- 8402655 TI - Homozygous deletions within 9p21-p22 identify a small critical region of chromosomal loss in human malignant mesotheliomas. AB - Previous DNA analyses have demonstrated that 9p13-p22 is a frequent site of chromosomal loss in leukemia, glioma, melanoma, and lung and bladder carcinomas. Recent cytogenetic studies have revealed recurrent alterations of 9p in malignant mesothelioma (MM). We have performed gene dosage studies of 23 MM cell lines, using probes for several 9p21-p22 loci (IFNB, IFNA/IFNW, D9S3, D9S126, D9S169, and D9S171), to identify a common region of deletion. Homozygous and/or hemizygous deletions were identified in 19 (83%) cell lines. Homozygous losses (10 cell lines; 43%) occurred most often at the D9S171 and IFNA/IFNW loci. In 8 cell lines, 2 or more of the 9p loci examined were found to be homozygously lost; 2 others displayed homozygous losses only at the D9S171 locus. Results from our deletion mapping analysis suggest that D9S171 is located between IFNA/IFNW and D9S126. The data presented here indicate that allelic loss from 9p21-p22 is a common occurrence in MM and further delineate the location of a putative 9p tumor suppressor gene(s) to a region between IFNA/IFNW and D9S171. These MM cell lines may facilitate efforts to define an even smaller critically deleted region, leading to the eventual cloning and characterization of this gene. PMID- 8402656 TI - Dexamethasone reduces the interstitial fluid pressure in a human colon adenocarcinoma xenograft. AB - The effect of dexamethasone on interstitial hypertension was evaluated in a human colonic adenocarcinoma. Two weeks after transplantation of the tumor line LS174T into SCID mice, recipients with tumors > 8.5 mm in diameter received one daily injection i.p. on days 1-4, at five different doses in the range of 0.3-30 mg/kg. Controls received saline. The interstitial fluid pressure (IFP) was determined in all tumors pretherapeutically on days 1, 4, and 7. A total of 68 tumors were examined, and in an additional group of 22 mice, the effect of 4-day dexamethasone therapy on blood pressure was evaluated. In the 3-, 10-, and 30 mg/kg dose groups a significant reduction in IFP was found, comparing treated versus controls and individual measurements from day 1 versus day 4. No effects were observed on day 7. A marginal effect was observed after 1.0 mg/kg, whereas 0.3 mg/kg did not affect the IFP. The systemic blood pressure was slightly increased by the dexamethasone therapy, and no treatment related changes in tumor sizes were observed. Our findings indicate that the reversible decrease in tumor IFP by dexamethasone is an effect of a reduced microvascular permeability and vascular hydraulic conductivity in the tumors. PMID- 8402657 TI - Prevention of carcinogenicity of anticancer drugs by metallothionein induction. AB - We examined the efficacy of metallothionein induction in the prevention of the carcinogenic action of cis-platinum and melphalan administered repeatedly to mice over a relatively long period. The increased pulmonary metallothionein induced by bismuth or zinc compounds during the period of chemotherapy with cis-platinum or melphalan protected the mice from carcinogenesis of these drugs in the lung. These results suggested the efficacy of metallothionein inducers in suppression of carcinogenicity considered as a secondary effect of anticancer agents in cancer chemotherapy. PMID- 8402658 TI - The synthetic retinoid fenretinide lowers plasma insulin-like growth factor I levels in breast cancer patients. AB - We studied the effect of fenretinide [N-(4-hydroxyphenyl)retinamide (4-HPR)], a synthetic analogue of retinoic acid, on plasma insulin-like growth factor I (IGF I) levels in a consecutive cohort of stage I breast cancer patients belonging to a randomized phase III trial of breast cancer chemoprevention. Thirty-two women receiving 4-HPR 200 mg/daily and 28 untreated controls entered the study. IGF-I levels were determined after acid-ethanol extraction, on plasma obtained at randomization and after a mean time of 10.8 +/- 0.3 months. At baseline, there was no difference in IGF-I levels between the two groups [152.9 +/- 9.4 versus 159.2 +/- 7.0 ng/ml in treated and control group (P = 0.59), respectively]. After follow-up time, while plasma IGF-I levels were unchanged in control patients (163.3 +/- 7.4 ng/ml; P = 0.5), they were significantly reduced to 134.6 +/- 8.1 ng/ml in the patients treated with 4-HPR (P = 0.003 and P = 0.011 versus baseline and control values, respectively). Multiple regression analysis showed that treatment was the only determinant of IGF-I decline. Moreover, the interaction between treatment and age was significant, in that the decrease of IGF-I levels induced by 4-HPR administration was much more pronounced in younger patients, while an age-related decline was observed in controls. We conclude that the synthetic retinoid 4-HPR lowers circulating IGF-I levels in early breast cancer patients. Although the importance of this observation for the clinical prevention of breast cancer remains to be established, it further substantiates the rationale of the combination of 4-HPR with tamoxifen, which is known to decrease IGF-I as well and to act synergistically with the retinoid in preclinical models. PMID- 8402659 TI - Inactive p53 mutants may enhance the transcriptional activity of wild-type p53. AB - The p53 mutants 248Trp, 175His, and 281Gly fail to activate transcription mediated by p53CON element (GGACATGCCCGGGCATGTCC) or the ribosomal gene cluster element (ACGTTTGCCTTGCCTGGACTTGCCTGGCCTTGCCTT). We studied the effect of these inactive p53 mutants on the transcriptional activity of wild-type p53 by cotransfection of both wild-type and mutant p53 expression vectors into p53-null K562 chronic myelogenous leukemia cells. The p53 mutants enhanced the p53CON mediated gene expression of wild-type p53 but decreased the wild-type p53 activated transcription mediated by ribosomal gene cluster. Thus, p53CON and ribosomal gene cluster represent distinct p53-binding elements. Furthermore, p53 mutants may affect the transcriptional activity of wild-type p53 in either a dominant positive or a dominant negative manner, depending on the binding element present. PMID- 8402660 TI - Role of the p53 tumor suppressor gene in cell cycle arrest and radiosensitivity of Burkitt's lymphoma cell lines. AB - We have assessed the role of the p53 tumor suppressor gene in cell cycle arrest and cytotoxicity of ionizing radiation in 17 Burkitt's lymphoma and lymphoblastoid cell lines. Cell cycle arrest was assessed by flow cytometry of cells 16 h following irradiation. In addition to the usual G2 arrest, the cell lines exhibited three types of responses in G1: Class I, strong arrest in G1 following radiation; Class II, minimal arrest; and Class III, an intermediate response. All Class I cells contained normal p53 genes. Of the ten lines that showed minimal G1 arrest, eight had mutant p53 alleles, and two lines were heterozygous for p53 mutations. Both of the lines showing an intermediate response contained wild-type p53. Our results are consistent with the view that mutations abrogate the ability of p53 to induce G1 arrest following radiation. Studies with the heterozygotes showed that the mutant protein can have a dominant negative influence upon wild-type p53, and the reduced ability of two normal p53 lines to arrest in G1 indicated that p53 function can be impaired by other mechanisms. The radiosensitivity of most of the lines appeared to depend on the ability of p53 to induce a G1 arrest. The mean radiation dose that inhibited proliferation of the Class I lines by 50% was 0.98 Gy. Of the eight p53 mutant cell lines tested, five lines required approximately 2.9 Gy to cause a 50% inhibition of cell proliferation. The two heterozygotes were also more resistant to radiation than the Class I cells (50% inhibitory dose, 2.1 and 2.9 Gy). Our results suggest that radioresistance is afforded by a loss of function of wild type p53, which would normally induce a G1 arrest and promote cell death in the presence of DNA damage. PMID- 8402661 TI - Maintenance of genomic imprinting at the IGF2 locus in hepatoblastoma. AB - Genomic imprinting is the parental allele specific expression of genes and has recently been shown to occur in humans. Evidence for a role for genomic imprinting in human cancer comes from the finding of preferential retention of paternal alleles in embryonal tumors undergoing loss of heterozygosity, e.g., Wilms' tumor and osteogenic sarcoma. Recent studies have demonstrated imprinting of the insulin-like growth factor II gene at 11p15 in normal individuals, with the paternally inherited allele expressed and the maternal allele silent. It has been shown that normal imprinting is relaxed, and gene expression is biallelic in a majority of Wilms' tumors which retain heterozygosity at this locus. In this study an intragenic ApaI polymorphism is used to examine imprinting of the insulin-like growth factor II gene in hepatoblastoma. Three of 5 tumors studied were heterozygous and hence informative. All cases showed monoallelic expression of the insulin-like growth factor II gene, indicating maintenance of normal imprinting at this locus. PMID- 8402662 TI - Molecular and cellular interactions between intoplicine, DNA, and topoisomerase II studied by surface-enhanced Raman scattering spectroscopy. AB - The surface-enhanced Raman scattering spectra of the new antitumoral agent, intoplicine (RP 60475, NSC 645008), and those of its complexes with DNA and topoisomerase II in vitro and in K562 cancer cells were obtained. Intoplicine was found to unwind DNA and to inhibit purified calf thymus topoisomerase II via a stabilization of the ternary cleavable complex. The intensity of the surface enhanced Raman scattering spectrum of intoplicine was not modified by the addition of plasmid pBR322 or calf thymus DNA. In the complex of this antitumor agent with topoisomerase II, the signal of intoplicine was completely abolished, indicating that at least some portion of intoplicine binds to an internal part of the enzyme. During the formation of the ternary complex, intoplicine was released from the interior of the protein and formed hydrogen bonds via its hydroxyl and/or amino groups. Similar modifications of the intoplicine spectra were found by microsurface-enhanced Raman scattering spectroscopy of the compound in the nucleus of treated K562 cells. In contrast, intoplicine was found to be in a free form in the cytoplasm. PMID- 8402663 TI - Monoclonal antibody 2B5 defines a truncated O-glycan, GlcNAc beta 1-3Gal beta 1 4GlcNAc beta 1-6 (GalNAc), on mucins from deep gastric and duodenal glands as well as metaplasia and neoplasia of gastric differentiation. AB - Monoclonal antibody (MAb) 2B5 previously generated in BALB/c mice using gastric mucosa of the corpus as immunogen was characterized with regard to its binding epitope. Binding assays on structurally defined neoglycolipids from mucin glycans revealed that MAb 2B5 recognizes a carbohydrate epitope on a neutral O-glycan sequence from gastric mucins. This oligosaccharide chain has been characterized by mass spectrometry and methylation analysis and sequential exoglycosidase treatment of the derived neoglycolipid as GlcNAc beta 1-3Gal beta 1-4GlcNAc beta 1-6OY (where 6OY corresponds to the GalNAc-ol fragment--O--(CH2)2 conjugated to dipalmitoylphosphatidylethanolamine). The selective binding of MAb 2B5 to mucus from deep gastric and duodenal glands, to gastric metaplasia or neoplasia with gastric differentiation may imply that mucin glycosylation in the respective cells is incomplete resulting in the accumulation of truncated blood group precursor chains. MAb 2B5 is the first monoclonal antibody which defines an epitope on M2-type antigens and may substitute, accordingly, for the inconvenient "paradoxical concanavalin A-horseradish peroxidase method" in histochemistry. PMID- 8402664 TI - Immortalization of Syrian hamster embryo cells: probabilistic event or deterministic process. AB - Previous findings on the induction of immortalization in SHE cells have been explained with the activation/alteration hypothesis which postulates that treatment with a carcinogen results in the induction of a so-called "activated state" which enhances the rate of a probabilistic event in the progeny of the treated cells. This event is supposed to be a mutation. Because it has been recently indicated that in mammalian cells the switching on of signal transduction pathways by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) or carcinogens can lead to genetic instability in the progeny of the treated cells, the possibility of an analogy between the induction of genetic instability and induction of immortalization after treatment with TPA was investigated. No effect of TPA was found on the rate of immortalization/cell/generation, not in otherwise untreated cells nor in cells treated with benzo(a)pyrene. TPA was found to enhance the life span of SHE cells. The life span of a culture correlated with its growth rate and its cell density at confluence both in the absence and presence of TPA. These correlations are supposed to reflect a regulation mechanism involved in the program of cellular senescence, and supposedly TPA can partly reverse this program. Treatment with benzo(a)pyrene also interferes with the life span resulting in premature senescence in most of the cells and extension of life span in a small fraction of the cells which subsequently can become immortal. Repeated switching from logarithmic growth to G0 also enhanced life span and rate of immortalization. The findings indicate that the activated state is a disturbance of a differentiation program affecting in SHE cells the program of cellular senescence and that, as an explanation for immortalization, epigenetic alterations causing a deterministic process of dedifferentiation in a subpopulation of the cells appear as plausible or perhaps even more plausible as a probabilistic mutation. This indicates that disturbance of differentiation might be among the causes of genetic instability. PMID- 8402665 TI - The suprabasal expression of alpha 6 beta 4 integrin is associated with a high risk for malignant progression in mouse skin carcinogenesis. AB - Enhanced expression of the alpha 6 beta 4 integrin complex has been linked to malignant progression in mouse skin carcinogenesis. To determine if alpha 6 beta 4 expression can predict risk for malignant conversion among populations of benign skin tumors, we analyzed the distribution of alpha 6 beta 4 and other markers of progression in papillomas at high and low risk for malignant progression. After initiation with 7,12-dimethylbenz[a]anthracene, mice were promoted with 12-O-tetradecanoylphorbol-13-acetate to induce predominantly low risk tumors or promoted with mezerein to produce predominantly high risk tumors. When tumors first appeared at 8 weeks after promotion, high risk papillomas demonstrated basal and suprabasal alpha 6 beta 4 expression, loss of keratin 1, and aberrant expression of keratin 13. In these tumors alpha 6 beta 4 expression coincided with an expansion of the proliferating compartment as indicated by suprabasal bromodeoxyuridine labeling. In contrast, alpha 6 beta 4 immunostaining was confined to basal cells in low risk tumors, keratin 1 was abundant, and keratin 13 was absent in the majority of this group, while proliferating cells were largely in the basal compartment. By 33 weeks, alpha 6 beta 4 suprabasal expression continued to distinguish groups at higher risk for malignant conversion, but keratin 13 was expressed in all groups. At this time, high risk tumors displayed focal expression of keratin 8 and gamma-glutamyltranspeptidase, markers also found in chemically induced carcinomas. Keratin 8 and gamma glutamyltranspeptidase were expressed only in alpha 6 beta 4 positive cells. These results indicate that expression of alpha 6 beta 4 integrin in suprabasal strata serves as an early predictive marker to identify benign squamous tumors at high risk for malignant progression. PMID- 8402666 TI - HPV-16, tobacco-specific N-nitrosamine, and N-methyl-N'-nitro-N-nitrosoguanidine in oral carcinogenesis. AB - We previously immortalized human oral keratinocytes by transfection with recombinant human papillomavirus type 16 (HPV-16) DNA and established two cell lines. These transfected cells were morphologically different from the normal counterpart, contained intact HPV-16 DNA in an integrated form, and expressed numerous viral genes. These cells contained lower levels of wild-type p53 protein and higher levels of c-myc mRNAs compared to normal cells. However, they proliferated only in keratinocyte growth medium containing a low level of calcium and were not tumorigenic in nude mice. A HPV-16-immortalized cell line was exposed to either 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone or N-methyl-N' nitro-N-nitrosoguanidine. Four chemically transformed cell colonies were isolated. These cells proliferated well in Dulbecco's minimum essential medium containing a physiological level of calcium. They contained, similar to the immortalized counterpart, integrated HPV-16 sequences and lower levels of both wild-type p53 protein and DCC messages compared to normal cells. Among the chemically transformed cells, two colonies obtained from 4-(methyl-nitrosamino)-1 (3-pyridyl)-1-butanone exposure demonstrated an enhanced proliferation capacity in nude mice and transcribed a substantially higher amount of HPV-16 E6/E7, epidermal growth factor receptors, and c-myc genes compared with the immortalized counterpart. These experiments indicate that malignant transformation of oral keratinocytes can be caused by a sequential combined effect of "high risk" HPV and tobacco-related carcinogens. PMID- 8402667 TI - p53 protein accumulates frequently in early bronchial neoplasia. AB - p53 mutations are common in human lung cancer and frequently generate levels of p53 protein that are detectable by immunohistochemistry. For this reason, p53 protein accumulation is a candidate biomarker, but little is known about its timing or frequency in multistage bronchial carcinogenesis. We studied human lung tissues containing preinvasive squamous neoplasms from 34 donors with and without lung cancer. Nuclear p53 protein was present in 0% of normal mucosas, 6.7% of squamous metaplasias, 29.5% of mild dysplasias, 26.9% of moderate dysplasias, 59.7% of severe dysplasias, 58.5% of carcinomas in situ, 67.5% of microinvasive carcinomas, and 79.5% of invasive tumors. These data indicate that (a) p53 protein accumulates in about 30% of the earliest recognized neoplastic lesions (i.e., mild dysplasia), (b) there is an increasing frequency of p53 protein accumulation starting with mild dysplasia, and (c) p53 protein accumulates infrequently in normal or metaplastic mucosa. In a subset of six patients whose most advanced lesion was carcinoma in situ without evidence of invasive cancer, p53 protein was detected in 0% of normal mucosas, 8.3% of squamous metaplasias, 37.5% of mild dysplasias, 12.5% of moderate dysplasias, 93.8% of severe dysplasias, and 55% of carcinoma in situ lesions. These data show clearly that p53 alterations can occur before invasion and suggest that the frequency is similar to that observed in the full series. Since two-thirds or more of lung cancers have p53 alterations, the timing and frequency of p53 protein accumulation make the p53 tumor suppressor gene an attractive marker for early diagnosis and evaluation of chemoprevention agents. PMID- 8402668 TI - High-risk human papillomavirus infections and overexpression of p53 protein as prognostic indicators in transitional cell carcinoma of the urinary bladder. AB - Ninety Japanese patients with transitional cell carcinoma of the urinary bladder were investigated for tumor incorporation of DNA for high-risk human papillomavirus (HPV) types 16, 18, and 33 by in situ hybridization with biotinylated DNA probes. In addition, immunohistochemical analysis of p53 protein expression was performed with an antibody to p53 protein. Twenty-eight tumors were positive for HPV DNA, and multiple HPV infection was detected in 17 cases. Positive nuclear staining of cancer cells by the antibody to p53 protein was detected in 32 cases. DNA for HPV 16, 18, and/or 33 and the overexpression of p53 protein were simultaneously observed in 6 tumors by using a mirror section method. The overexpression of p53 protein was frequently detected in invasive and nonpapillary tumors (P < 0.05) and in high grade tumors (P < 0.05). In contrast, HPV infection was more common in noninvasive and papillary tumors (P < 0.01). The patients with tumors positive for HPV DNA and/or p53 antibody had a significantly worse survival rate (P < 0.05). These results suggest that HPV infection or overexpression of p53 protein may be related to tumor behavior and may indicate a relatively poor prognosis in patients with transitional cell carcinoma. PMID- 8402669 TI - A phase I study of continuous infusion 5-fluorouracil plus calcium leucovorin in combination with N-(phosphonacetyl)-L-aspartate in metastatic gastrointestinal adenocarcinoma. AB - Preclinical studies suggest that the biochemical effects of N-(phosphonacetyl)-L aspartate (PALA), an inhibitor of aspartate carbamoyltransferase (ACTase), may increase the metabolic activation of 5-fluorouracil (5-FU) and enhance its cytotoxicity through both RNA- and DNA-directed mechanisms. In this Phase I trial, 22 evaluable patients with adenocarcinoma of the gastrointestinal tract were entered at escalating doses of 5-FU starting at 1150 mg/m2/day given as a concurrent 72-h i.v. infusion with a fixed dose of leucovorin (LCV), 500 mg/m2/day. The dose of 5-FU was escalated within patients according to individual tolerance, and then PALA at 250 mg/m2 was added 24 h prior to the initiation of the 5-FU/LCV infusion of the subsequent cycle. Dose-limiting mucositis and myelosuppression occurred during the initial cycle in 3 of 5 patients treated with 2300 mg/m2/day 5-FU; therefore, the recommended dose of 5-FU with concurrent LCV is 2000 mg/m2/day. Twenty-seven additional patients were then treated with escalating doses of PALA ranging from 375 to 2848 mg/m2, i.v., followed 24 h later by 2000 mg/m2/day 5-FU with high-dose LCV. Dose-limiting mucositis and myelosuppression occurred during the initial cycle in 2 of 3 patients entered at 2848 mg/m2 PALA. Dose-limiting mucositis and skin rash ultimately required both PALA and 5-FU dose reductions in 4 of 6 patients treated with 1899 mg/m2 PALA. Toxicity was similar, however, in patients receiving PALA at doses ranging from 375 to 1266 mg/m2. The mean steady-state plasma concentration of 5-FU at 2000 mg/m2/day was 6.5 +/- 0.9 microM; patients with 5-FU levels > 9 microM had a significantly higher incidence of serious gastrointestinal and hematological toxicity. Compared to each patient's own baseline, a significant trend for decreasing ACTase activity with increasing PALA dose was evident using cytosol isolated from peripheral blood mononuclear cells 24 h after PALA treatment (P2 = 0.01). PALA < or = 844 mg/m2 failed to appreciably inhibit ACTase activity at 24 h in most patients; furthermore, a decrease in ACTase activity by > 50% from baseline was seen in only 29% of cycles. More consistent inhibition of ACTase activity was seen with PALA > or = 1266 mg/m2. Even with the highest PALA doses, however, ACTase activity returned to baseline by 96 h in most patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402670 TI - Phase I pharmacokinetic study of cyclosporin A combined with doxorubicin. AB - We performed a phase I trial of cyclosporin A (CsA) in combination with doxorubicin (dox) to determine the maximally tolerated dose (MTD) of the combination in man, to define the quantitative and qualitative toxicities of the combination, and to determine the pharmacokinetics of the two drugs when used together. CsA was administered as a continuous infusion for 6 days, and dox was administered as a single 10-min infusion 24 h after the initiation of CsA. The starting CsA infusion rate was 5 micrograms/kg/min, and the dox starting dose was 30 mg/m2. Courses were administered every 4 weeks with first CsA and then dox being escalated in consecutive cohorts of patients until the MTD was determined. Twenty-three patients and 40 courses were evaluable for toxicity. Pharmacokinetic analysis was performed in 23 patients on the first course for whole blood CsA and plasma dox and doxorubicinol. The MTD of CsA was 6 micrograms/kg/min, and for dox it was 45 mg/m2. Dose-limiting toxicity was neutropenia. Serum creatinine and creatinine clearance did not change over the infusion period. Bilirubin increased from a median of 10 mumol/liter at the initiation of the infusion to a median of 40.4 mumol/liter at the end of the infusion but returned to normal before the next cycle of therapy. Nausea and vomiting were common and marked, whereas thrombocytopenia was mild. Two patients, one with small cell lung cancer and one with breast cancer, had stable disease while receiving treatment for 5 and 6 months, respectively. Mean whole blood steady state concentrations of CsA were 2210 ng/ml during the infusion with total body clearance of 0.177 liter/h/kg. The area under the concentration x time curve (AUC) increased linearly with dose of dox, and total body clearance was independent of dose. The mean total body clearance was 2.46 liters/h/m2, and terminal half-life was 49.6 h. The AUC for dox was greater and clearance was less than has been previously reported at the doses administered in this study. The ratio of AUC for doxorubicinol to AUC for dox was less than expected, suggesting that the metabolism and/or excretion of dox was decreased when administered with CsA. We conclude that dox can be combined with infusioned CsA but at a lower dose than when given alone. This may be due to altered metabolism and/or excretion of dox or increased bone marrow stem cell sensitivity to dox. PMID- 8402671 TI - Phase I evaluation and pharmacokinetic study of pyrazine-2-diazohydroxide administered as a single bolus intravenous injection in patients with advanced solid tumors. AB - The sodium salt of pyrazine-2-diazohydroxide (PZDH; NSC 361456) was identified as an active congener of the antitumor lead pyridine-2-diazotate with enhanced chemical stability under physiological conditions. In a phase I trial of PZDH administered as a single i.v. bolus injection, 19 patients with refractory solid tumors received 44 courses of therapy at dose levels ranging from 50 to 350 mg/m2. No objective responses to PZDH were noted. Myelosuppression characterized by prolonged, delayed onset leukopenia and thrombocytopenia was the dose limiting toxicity. A maximum tolerated dose of 350 mg/m2 was identified for this treatment schedule. Nonhematological toxicity was limited to severe nausea and vomiting, experienced by all patients treated at the lower doses, although reasonably well controlled when antiemetics were given prior to chemotherapy. The plasma pharmacokinetics of PZDH was evaluated following a single course of therapy in 16 patients. Drug levels were monitored using a specific capillary gas chromatographic assay with a 1-ng/ml lower limit of quantitation. In patients treated with doses greater than 50 mg/m2, the concentration of PZDH in plasma declined in a distinctly triexponential manner and remained above 1.5 ng/ml for at least 8 h. However, the initial decay phase, characterized by a harmonic mean half-life of 3.9 +/- 3.5 (SD) min (range, 2.2-6.3 min), was the primary determinant of drug disposition, as indicated by its 85.5-93.1% contribution to the area under the plasma concentration-time profiles from time zero to infinity. The harmonic mean terminal half-life increased with escalations in dose from 2.7 +/- 0.8 h (n = 2) at 100 mg/m2 to 8.5 +/- 3.0 h at 350 mg/m2 (n = 6). Total plasma drug clearance was very similar in patients treated with doses of 50-250 mg/m2, exhibiting a mean value of 42.5 +/- 7.8 liters/h/m2 (n = 10); however, it was significantly lower at the 350 mg/m2 dose level, 27.2 +/- 6.6 liters/h/m2 (n = 6; P < 0.002), denoting a departure from linear pharmacokinetic behavior. The rather low steady state apparent volume of distribution, which ranged from 6.0 +/ 1.5 (50 mg/m2, n = 2) to 12.7 +/- 8.0 (350 mg/m2, n = 6) liters/m2, was indicative of limited distribution of the drug into body tissue. The absence of objective antitumor effects should not discourage continued evaluation of PZDH against solid tumors selected for probable sensitivity as indicated by preclinical testing. A dose of 250 mg/m2 on a single i.v. bolus schedule is recommended for these phase II trials. PMID- 8402672 TI - Plasma and cellular pharmacokinetics of mitoxantrone in high-dose chemotherapeutic regimen for refractory lymphomas. AB - We have studied the plasma and peripheral leukocyte pharmacokinetics of mitoxantrone associated, in a high-dose regimen, with cyclophosphamide, carmustine, and etoposide in patients with refractory lymphoma undergoing autologous bone marrow transplantation. Nineteen patients with refractory lymphoma were involved in a clinical trial with escalated doses (15-90 mg/m2) of mitoxantrone administered by 30-min i.v. infusion 8 days before an autologous bone marrow transfusion. Mitoxantrone was measured by high-performance liquid chromatography in plasma and peripheral lymphocytes. The plasma pharmacokinetics of mitoxantrone was linear between 15 and 90 mg/m2: total body clearance (19.3 +/ 6.2 liter/h/m2) and volume of distribution at steady state (486 +/- 254 liter/m2) were not altered by increasing the dose. The exposure of bone marrow transplant to the plasma residual concentration of mitoxantrone was correlated with the limiting hematological toxicity of the regimen (P < 0.001). At all times, the mitoxantrone concentration in peripheral cells was much higher than in plasma and was retained at a constantly high concentration level. Whereas cellular versus plasma maximum concentration ratio was near 1, the area under the concentration +/- time curve ratio reached 100, suggesting a long elimination half-life from cells. Plasma and cellular pharmacokinetics data of mitoxantrone reinforce the idea that this drug is a good candidate for high-dose chemotherapy regimen. PMID- 8402673 TI - Human prostatic inhibin suppresses tumor growth and inhibits clonogenic cell survival of a model prostatic adenocarcinoma, the Dunning R3327G rat tumor. AB - Prostatic inhibin (PI) is a M(r) 10,700 protein found in human seminal plasma and is secreted by the prostate. Recognition of alteration of PI levels in prostatic diseases prompted us to investigate its effect on an animal prostatic adenocarcinoma model, the Dunning R3327G rat tumor. PI not only inhibited in vitro growth of tumor cells but also suppressed tumor growth in vivo. A dose dependent inhibition of both the clonogenic cell growth and rate of proliferation (DNA synthesis) was observed in tumor cell cultures incubated with purified PI. These inhibitory activities were similar in both androgen-dependent and androgen independent Dunning tumor cell lines. A functional decapeptide of PI was also found to inhibit Dunning tumor cell colonies in a dose-dependent manner. Daily injection of purified PI into tumor-bearing rats suppressed the tumor growth. A 58% reduction in tumor weight and a 2-fold reduction in tumor growth rate were observed over a 15-day treatment period. Continued treatment with PI significantly suppressed the tumor growth rate by nearly 3-fold. These findings clearly demonstrate a potential application of PI for treating human prostatic adenocarcinoma. PMID- 8402674 TI - A prospective cohort study on selenium status and the risk of lung cancer. AB - Selenium has been suggested to be anticarcinogenic and to play a role in the cellular defense against oxidative stress. The association between toenail selenium (a marker of long-term selenium status) and lung cancer was investigated in a cohort study of diet and cancer that started in 1986 among 120,852 Dutch men and women aged 55-69 years. After 3.3 years of follow-up, 550 incident cases of lung carcinoma were detected. Toenail selenium data were available for 370 lung cancer cases and 2459 members of a randomly selected subcohort. The rate ratio of lung cancer for subjects in the highest compared to the lowest quintile of toenail selenium, after controlling for age, gender, smoking, and education, was 0.50 (95% confidence interval, 0.30-0.81), with a significant inverse trend across quintiles (P = 0.006). The protective effect of selenium was concentrated in subjects with a relatively low dietary intake of beta-carotene or vitamin C. The rate ratio in the highest compared to the lowest quintile of selenium was 0.45 in the low beta-carotene group (95% confidence interval, 0.22-0.92; trend P = 0.028) and 0.36 in the low vitamin C group (95% confidence interval, 0.17-0.75; trend P < 0.001). The results of this study support an inverse association between selenium status and lung cancer and suggest a modification of the effect of selenium by the antioxidants beta-carotene and vitamin C. PMID- 8402675 TI - Metallothionein: a protein conferring resistance in vitro to tumor necrosis factor. AB - The role of metallothionein (MT) in the cytotoxicity of tumor necrosis factor (TNF) was investigated in vitro. A human epithelial cell line (HE100) and a mouse fibroblast line (CI 1D100) had previously been cultured to become resistant to 100 microns CdCl2 and were cultured routinely in cadmium-containing medium. The MT content of these cells and the nonresistant parent cell lines (HE and CI 1D) was determined both qualitatively and quantitatively by immunochemical techniques. Immunofluorescence microscopy revealed the cadmium-resistant cell lines to be intensely rich in MT in both the nuclear and cytoplasmic compartments. This finding was confirmed by immuno-electron microscopy which also showed the labeling to be freely distributed and not membrane-bound. In comparison, a very weak labeling of the parent cell lines was observed. MT concentration, as determined by enzyme-linked immunosorbant assay, was found to be 4.05 +/- 1.13% and 3.91 +/- 0.7% of the total protein for HE100 and CI 1D100 cells, respectively. We were unable to detect MT in the parent cell lines by this technique. Dose-survival curves obtained after 3 days treatment of the cells with TNF (0.125-500 ng/ml) revealed that the MT-rich substrains were significantly resistant compared to the parent strains (P < 0.001; t tests). In growth rate studies, where cells were exposed to TNF over a dose range of 0.25-250 ng/ml for 6 days, the resistance of the MT-rich cell lines was confirmed (P < 0.002). These data indicate that MT confers resistance to the cytotoxic effects of TNF in vitro and that sensitivity to TNF may be related to the MT content of the cell. PMID- 8402676 TI - Etoposide pharmacokinetics in children: the development and prospective validation of a dosing equation. AB - Pharmacokinetic studies of etoposide administered at 100-200 mg/m2 to 33 children are described. Twenty-seven studies were performed in children aged < 10 years. Repeat studies were performed in 11 patients. Median pharmacokinetic parameters were as follows: plasma clearance, 26 ml/min/m2; volume of distribution, 4.9 liters/m2; area under the etoposide plasma concentration-time curve (AUC), 3.9 mg/ml x min per 100 mg/m2. Interindividual variability in pharmacokinetic parameters was large (coefficient of variation (CV) = 30, 28, and 27%, respectively) in comparison with intraindividual variability (CV = 12, 14, and 12%, respectively). Variability in AUC was much greater in those patients treated with 150-200 mg/m2 etoposide than with 100 mg/m2 (CV, 35 versus 13%) and was related to variability in renal function and prior exposure to cisplatin. Data from the first 20 studies were used to develop pharmacokinetic monitoring equations which were validated in a further 13 patients. The most accurate equation relies upon the elimination constant of 51Cr-EDTA and a single blood specimen taken at the end of the etoposide infusion. [formula: see text] where K = 51Cr-EDTA elimination rate constant. This equation showed no significant bias, and the predictive error was small with respect to AUC calculated according to a two-compartment model. Predictive error did not increase with increasing AUC, whereas a marked increase in predictive error was seen for dosing according to body surface area. Dosing according to body surface area alone led to marked over or underexposure to etoposide in 8 patients. Pharmacokinetic monitoring using the equation described would have identified these patients and permitted dose modification. This approach provides an accurate means of monitoring etoposide AUC for administration times of 1-4 h without the need for detailed pharmacokinetic sampling. It will allow a significant reduction in the variability of exposure seen with surface area-based dosing. PMID- 8402677 TI - Development of a novel spontaneous metastasis model of human osteosarcoma transplanted orthotopically into bone of athymic mice. AB - There is a pressing need for in vivo models in which potential antitumor agents can be tested for their ability to inhibit the growth and metastatic spread of human sarcomas. A recent advance in this regard has been the development of a v Ki-ras-oncogene-transformed human osteosarcoma cell line (KRIB) that efficiently colonizes the lungs of athymic nude mice when cells (1 x 10(5)) are administered by i.v. injection. In the present study, we have utilized this cell line to develop a spontaneous metastasis model in which a small number of tumor cells are injected into the tibial bones of athymic mice. When as few as 1000 KRIB cells are orthotopically implanted into the tibial bones of nude mice, bone tumors, which are radiographically and histologically similar to primary human osteosarcoma, develop within 4 weeks. Furthermore, as in the human disease, cells from these primary tumors subsequently seed the animals' lungs, resulting in reproducible and quantifiable pulmonary metastasis evident both upon gross inspection of the lungs and histologically 6 weeks after tumor inoculation. Surgical amputation of the tumor inoculation site up to 2 weeks after tumor injection prevents pulmonary metastasis, indicating that substantial local (tibial) growth and invasion of the primary tumor for at least 2 weeks is required for subsequent metastasis. Implantation of s.c. 5000 KRIB cells fails to produce local or metastatic tumors. We anticipate that this model will prove to be a powerful tool with which to study the mechanisms of human osteosarcoma growth and pulmonary metastasis, and to assess the efficacy of promising therapeutic agents. PMID- 8402678 TI - Inhibition of experimental metastasis by an alpha-glucosidase inhibitor, 1,6-epi cyclophellitol. AB - Isolated from a culture filtrate of Phellinus sp., cyclophellitol is a specific inhibitor of beta-glucosidase, but unlike castanospermine, it does not inhibit experimental metastasis. However, its structural analogue, 1,6-epi cyclophellitol, inhibited alpha-glucosidase as well as beta-glucosidase, and inhibited experimental metastasis. 1,6-Epi-cyclophellitol depressed alpha glucosidase activity in cultured B16/F10 cells after 48 h of incubation. Preincubation of B16/F10 cells for 48 h with 1,6-epi-cyclophellitol inhibited invasion of the cells in a Boyden chamber assay at the doses effective in inhibiting alpha-glucosidase in situ. Pulmonary metastasis of B16/F10 cells in mice was inhibited by pretreatment of the cells with 1,6-epi-cyclophellitol in culture. The inhibitor reduced the collagen type I- and IV-mediated attachment of the cells, whereas it had no effect on laminin-mediated attachment. These results suggest that alpha-glucosidase in tumor cells is essential for the metastatic process through the cellular interaction with collagen type I and IV. PMID- 8402679 TI - Expression of a rat glutathione-S-transferase complementary DNA in rat mammary carcinoma cells: impact upon alkylator-induced toxicity. AB - The role of glutathione-S-transferase (GST) in alkylator drug resistance has been studied in MatB rat mammary carcinoma cells. A series of GST transfectant cell lines was established by using an expression vector containing the complementary DNA for the rat GST Yc gene under regulation of the SV40 early region promoter and the antibiotic resistance plasmid pSV2neo. Transfectant cell lines expressing up to 4-fold higher total GST activity than in the parental wild type cell line were identified. Southern blot analysis confirmed a DNA fragment corresponding in size to the transfected GST Yc complementary DNA. Wild type MatB cells contain very low levels of Yc protein, whereas the Yc+ clones showed greatly increased amounts of the Yc subunit. The effect of increased GST Yc activity on the sensitivity of the transfected clones to various cytotoxic agents was assessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cell survival assay. The clones expressing recombinant GST Yc were more resistant to melphalan (6- to 12-fold), mechlorethamine (10- to 16-fold), and chlorambucil (7- to 30-fold). In late passage populations of the GST Yc+ clones that had been grown over a period of 14 months under continuous selection in G418, GST activity was decreased and it was paralleled by a decrease in Yc protein. These late passage clones with diminished GST Yc content also demonstrate a partial reversion toward the wild type phenotype as determined by cytotoxicity assays using melphalan, mustargen, and chlorambucil. Interstrand DNA cross-links induced by mechlorethamine were significantly lower at 0, 2, and 20 h posttreatment in one of the GST Yc+ clones when compared to wild type MatB cells. These studies indicate that GST Yc overexpression can confer resistance to alkylating agents and that this correlates with inhibition of DNA cross-link formation. PMID- 8402680 TI - Reductive activation of mitomycin C by NADH:cytochrome b5 reductase. AB - Mitomycin C requires bioreduction in order to exert its cytotoxic action. Activation of mitomycin C to an electrophile was equally supported by NADPH and NADH in EMT6 tumor cell sonicates under hypoxia. No alkylation was observed under aerobic conditions. Purified NADH:cytochrome b5 reductase catalyzed the reduction of mitomycin C with a Km of 23 microM at pH 6.6. At pH 7.6, the rate of enzymatic reduction of mitomycin C was 61% of that at pH 6.6. NADH:cytochrome b5 reductase catalyzed the activation of mitomycin C to alkylating metabolites under both hypoxic and aerobic conditions, with alkylation being 1.5 times greater in hypoxia. Dicumarol at 100 microM inhibited the NADH:cytochrome b5 reductase catalyzed reduction of mitomycin C by 24% and by 57% at 300 microM. The degree of inhibition of the enzyme by dicumarol was the same at both pH 6.6 and 7.6. NADH:cytochrome b5 reductase exhibited a small but measurable NADH-oxidase activity, which was unaffected by 300 microM dicumarol. These findings demonstrate that (a) NADH:cytochrome b5 reductase can metabolically activate mitomycin C and (b) dicumarol is capable of inhibiting this enzymatic activity. PMID- 8402681 TI - Cellular pharmacology of liposomal cis-bis-neodecanoato-trans-R,R-1,2 diaminocyclohexaneplatinum(II) in A2780/S and A2780/PDD cells. AB - We studied the cytotoxicity, cellular accumulation, and DNA interactions induced by liposome-entrapped NDDP (L-NDDP) and cisplatin in A2780 human ovarian carcinoma cells sensitive (A2780/S) and resistant (A2780/PDD) to cisplatin. L NDDP was 2-fold more cytotoxic than cisplatin against A2780/S cells with 5-h or 24-h drug exposure. Both cell lines were equally sensitive to L-NDDP, while A2780/PDD cells were 4-fold resistant to cisplatin (resistance indexes, 1.30-1.85 and 4.02-4.50, respectively). Using a drug exposure time of 24 h, L-NDDP cytotoxicity was independent of the liposome composition used, whereas with shorter drug exposure (5 h), the cytotoxicity of L-NDDP was directly related to the relative content of DMPG in the liposome carrier. However, changes in liposome composition or drug exposure time did not alter the resistance index of L-NDDP in this cell system. The cellular accumulation of L-NDDP was similar in both cell lines and 2- to 5-fold higher than that of cisplatin in A2780/S cells, whereas the cellular accumulation of cisplatin was reduced by 2- to 3-fold in A2780/PDD cells. The presence of DMPG in the lipid bilayer enhanced by 2-fold the cellular accumulation of L-NDDP, in good correlation with the direct relation between cytotoxic potency of L-NDDP and the presence of DMPG in the liposome carrier. Pt/DNA levels were determined at different time points after drug exposure for 1 h. Peak Pt/DNA levels were observed at 6 h for cisplatin and at 9 h for L-NDDP. Peak Pt/DNA levels and Pt/DNA over time of L-NDDP were about 1.5- and 3-fold higher than those of cisplatin in A2780/S and A2780/PDD cells, respectively, when equimolar concentrations of both drugs were used. Cisplatin induced significant DNA interstrand and DNA-protein cross-links in A2780/S cells, and a good correlation was observed between cytotoxicity against both cell lines and both types of lesions. In contrast, equimolar concentrations of L-NDDP induced only minimal DNA interstrand cross-links in either cell line.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402682 TI - Expression of a recombinant breast tumor-associated mucin fusion protein in Escherichia coli exposes the tumor-specific epitope. AB - Mucins are highly immunogenic glycoproteins that are abundantly expressed by breast carcinomas and other carcinomas. The fact that deglycosylated normal mucin can induce tumor-specific monoclonal antibodies indicates that tumor-specific epitopes are hidden in the fully glycosylated form. Using recombinant DNA techniques, a fragment of mucin tandem repeats was inserted into pMal-p, an Escherichia coli expression vector, and resulted in the expression of an unglycosylated maltose-binding protein-mucin fusion protein. This fusion protein has been purified and showed strong affinity to breast tumor-specific monoclonal antibody SM3. The antisera against this recombinant mucin fusion protein recognized all breast tumor cell lines we tested. Competition assay with monoclonal antibody SM3 shows that anti-recombinant mucin fusion protein binds the epitope that SM3 binds. These results confirm the hypothesis that unglycosylated mucin contains a tumor-specific epitope. This leads to the possibility that recombinant mucin may be used to develop vaccines against breast cancer and cytotoxic T-lymphocyte lines for immunotherapy of breast cancer. PMID- 8402683 TI - Human cytotoxic T-cells suppress the growth of spontaneous melanoma metastases in SCID/hu mice. AB - Mice with severe combined immunodeficiency (scid) provide an excellent model for studying interactions between human tumor cells and effector cells of the immune system. Because these animals lack functional B and T lymphocytes, they can accept human tumor xenografts and transfer of human effector cells. Here, we determined the ability of a human melanoma-specific, cytotoxic T-cell line (CTL) in suppressing the growth of spontaneously metastasizing human melanoma cells M24 met (HLA-A11, A33) in scid mice. This CTL line was highly cytotoxic and restricted by HLA-A11 against M24 met melanoma cells in vitro but poorly cytotoxic when tested against a human melanoma cell line that did not express HLA A11. In order to evaluate the efficacy of this CTL line against M24 met melanoma cells in vivo, randomized groups of animals were given injections of either RPMI culture medium, interleukin 2 (IL-2), CTLs, or CTLs + IL-2. IL-2, per se, did not significantly reduce tumor metastases; however, injection of melanoma-specific, HLA-A11 restricted CTLs into scid mice, 1 day postexcision of the previously induced primary tumor, markedly reduced the number of metastatic foci in the lung and decreased metastatic involvement in lymph nodes. The combination of these CTLs with IL-2 proved even more effective, since almost all lung metastases were eradicated and metastatic involvement in both axillary and inguinal lymph nodes was substantially reduced. Our results indicate that these human CTLs maintain their ability for specific killing of metastasizing melanoma cells in scid mice. Our data suggest that reconstitution of scid mice with a specific group of effector cells (step-wise scid/hu) may be helpful for in vivo evaluation of potentially useful cancer immunotherapy modalities. PMID- 8402684 TI - The expression of mouse biliary glycoprotein, a carcinoembryonic antigen-related gene, is down-regulated in malignant mouse tissues. AB - Mouse biliary glycoprotein (Bgp) is a member of the carcinoembryonic antigen gene family and is highly expressed in the epithelial cells of normal hepatic biliary ducts and intestine. Nine mouse Bgp isoforms have been identified through molecular cloning and shown to be splice and allelic variants of one Bgp gene. These glycoproteins function in vitro as intercellular adhesion molecules and serve as the mouse hepatitis virus receptors. Since human carcinoembryonic antigen is overexpressed in gastrointestinal tumors, we have investigated the expression of mouse Bgp in primary tumors and carcinoma cell lines. Our results demonstrate that the expression of the major mouse Bgp isoforms is down-regulated in tumors at the transcriptional and the posttranscriptional levels. This decrease in expression is corroborated by immunostaining of primary colonic tumors with anti-mouse Bgp antibodies. In addition, Bgp expression is influenced by transcriptional control mechanisms involving DNA methylation of the Bgp gene upstream regulatory region. Our results demonstrate that mouse Bgp protein expression is decreased upon malignant transformation and further suggest that Bgp proteins may be involved in the maintenance of the differentiated cellular phenotype. PMID- 8402685 TI - Multiple mechanisms of N-(phosphonoacetyl)-L-aspartate drug resistance in SV40 infected precrisis human fibroblasts. AB - Normal and SV40-infected human fibroblasts were grown in the presence of the drug N-(phosphonoacetyl)-L-aspartate (PALA) and examined for evidence of genetic instability. Both cell populations were precrisis and showed a normal, diploid karyotype at early passage. In contrast to the normal IMR-90 cells, which showed growth arrest and did not form colonies in PALA, the SV40-infected IMR-90 cells formed colonies at a very high frequency and continued to cycle in the drug. The drug-resistant colonies senesced after continued growth in culture, indicating that this change in ability to amplify preceded immortalization. This is the first observation of mortal human cells overcoming the drug-induced growth arrest. Although all previously isolated PALA-resistant colonies demonstrated CAD gene amplification as the mechanism of the drug-resistant phenotype, these SV40 infected human cells also showed alternative mechanisms, including increases in gene copy number by aneuploidy and formation of an isochromosome 2p. PMID- 8402686 TI - Hyperthermia induces resistance to ultraviolet light B in primary and immortalized epidermal keratinocytes. AB - Environmental exposure to UVB (290-320 nm) wavelengths of the solar spectrum causes major damage, including carcinogenesis, in the skin. Therefore, cellular responses that protect against UVB damage are of particular interest in cutaneous epithelial cells. In cultured keratinocytes, mild hyperthermia generates a classical stress response with acquired thermotolerance and elevated stress protein synthesis (E. V. Maytin, J. Biol. Chem., 267: 23189-23196, 1992). To test the ability of this stress response to protect against UVB damage, monolayers of primary murine keratinocytes or BALB/MK keratinocytes were heated at 42 degrees C for 1 h and then exposed to UVB at 6 h (typical dose, 40 mJ/cm2). Survival was assessed by fluorescein diacetate/ethidium bromide vital dye uptake and video microscopy. With heat-conditioning prior to UVB, a significant increase in both the percentage viability (2- to 3-fold) and in the absolute number of living (fluorescein diacetate-positive) cells was measurable at 24-48 h. Steady-state incorporation into [3H]DNA and 35S-protein, while suppressed immediately after UVB, showed greater recovery in heat-conditioned cultures compared to sham conditioned cultures at 48 h. Increased metabolic activity was accompanied by increased proliferative potential since colonies of BALB/MK cells observed at 72 h were larger, more numerous, and more active in the uptake of 5-bromo-2' deoxyuridine in heat-conditioned cultures. A time course for the development of UVB resistance showed maximal protection when heat and UVB were spaced approximately 6 h apart. Hyperthermic conditioning could induce UVB protection in nonproliferating cells, indicating that cell cycle arrest was not primarily responsible for the UVB-protective effect. In summary, hyperthermia induces a mechanism in epithelial cells which can ameliorate damage from UVB. PMID- 8402687 TI - Altered extracellular matrices influence cellular processes and nuclear matrix organizations of overlying human bladder urothelial cells. AB - A central issue in tumor biology is the understanding of the interactions between tumor cells and their environment. Using normal and ras oncogene transfected rat fibroblast cells, we now demonstrate that the transfected cells make altered extracellular matrices (ECM) and that their resulting ECM influence the proliferation and genetic regulation of human bladder cancer EJ cells. Using Western blot analyses, we observed that the ras transfected fibroblast cells lacked the ability to produce extracellular matrix component laminin whereas the normal parental fibroblast cells were able to produce intact laminin. Both transfected and nontransfected fibroblast cells were able to synthesize other extracellular matrix molecules such as type IV collagen and fibronectin. Human bladder tumor EJ cells were grown on ECM derived from normal and transfected rat fibroblast cells, and the proliferation rate and type IV collagen mRNA expression of EJ cells were determined. We observed that EJ cells, when grown on ECM derived from the ras transfected fibroblast cells, had a higher growth rate than when grown on ECM derived from the normal fibroblast cells (P < 0.037). Furthermore, EJ cells grown on ECM derived from transfected fibroblast cells showed up regulation of type IV collagen mRNA expression when compared with EJ cells grown on ECM derived from nontransfected fibroblast cells. Finally EJ cells grown on purified laminin but not on collagen IV coated flasks showed the same level of type IV collagen mRNA expression as when grown on ECM derived from nontransfected parental fibroblast cells. Haptotactic/motility assays with EJ cells and ECM derived from ras transfected and nontransfected fibroblast cells demonstrated that ECM of ras transfected fibroblast cells, but not the parental fibroblast cells, provided a permissive or fertile soil for EJ tumor cell invasion. Finally, two-dimensional gel electrophoresis of 35S-labeled nuclear matrix proteins of EJ cells cultured on ECM derived from ras transfected fibroblast cells revealed expression of proteins in the molecular weight range of M(r) 35,000-45,000 and isoelectric focusing pH range of 5.5 to 6.0. These proteins were not present in EJ cells cultured on ECM derived from parental nontransfected fibroblast cells. We conclude that extracellular matrices derived from transformed stroma producing cells may influence the proliferation, genetic regulation, and maintenance of the overlying urothelial tumor cells. The mechanism by which the ECM may influence cellular behavior and phenotype may be in their ability to modulate the nuclear matrix proteins of the overlying cell. PMID- 8402688 TI - Regulation of Bcl-2 oncoprotein levels with differentiation of human neuroblastoma cells. AB - When established in culture, human neuroblastoma cell lines typically are comprised of heterogeneous cellular subpopulations, including neuroblastic (N type), substrate-adherent (S-type), and intermediate (I-type) cells that can be distinguished by their characteristic morphologies and expression of differentiation-associated antigens. Here we examined the relative levels of the Bcl-2 oncoprotein in 15 clones derived from four different neuroblastoma cell lines. Among six clones isolated from the SK-N-SH line, levels of p26-Bcl-2 correlated with morphology and differentiation markers with the hierarchy of bcl 2 expression being: N-type cells > N/I-type > I-type > S-type. Furthermore, stimulation of one of the N-type clones, SH-SY5Y, with the phorbol ester, 12-O tetradecanoylphorbol-13-acetate, induced differentiation toward a more neuronal like phenotype and resulted in a 5- to 10-fold elevation in the relative levels of Bcl-2 protein. High relative amounts of p26-Bcl-2 protein were also found in an N-type clone derived from the SMS-KCN line. In two N-type clones derived from the LA-N-1 line, however, levels of Bcl-2 protein were only moderately elevated, and in one N-type clone from the SK-N-BE(2) line the levels of Bcl-2 protein were low. Thus, high relative levels of Bcl-2 oncoprotein are not a universal feature of N-type cells (three of six clones tested). In contrast, all 5 of the S-type clones evaluated contained relatively low levels of Bcl-2 protein, suggesting that these cells (which may represent embryonic precursors of Schwann, glial, and melanocytic cells) do not typically express the bcl-2 gene at high levels. Consistent with this inverse correlation between Bcl-2 protein levels and S-type characteristics, stimulation of an I-type clone derived from the SK-N-BE(2) line with 5-bromodeoxyuridine was accompanied by an accumulation of S-type cells in these cultures, decreased Bcl-2 protein, diminutions in the neuronal markers neurofilament-M and neuron-specific enolase, and an increase in the relative levels of the S-type marker proteins vimentin and beta-2-microglobulin. Conversely, stimulation of this I-type clone with retinoic acid resulted in an accumulation of N-type cells (which are thought to represent embryonic precursors of sympathetic neurons), decreased vimentin and beta-2-microglobulin, increased neurofilament-M, and a marked elevation in p26-Bcl-2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402689 TI - E-cadherin: a differentiation marker in thyroid malignancies. AB - Loss of E-cadherin (uvomorulin), a Ca(2+)-dependent cell adhesion molecule required for normal epithelial function, has been attributed a pathogenetic role in tumor invasion. The expression of E-cadherin was studied in normal and neoplastic follicular epithelium of the human thyroid by Northern blot analysis and immunofluorescence on frozen tissue sections. In the normal thyroid (n = 10) and in benign thyroid disorders (n = 21; toxic diffuse goitre; multinodular goitre; follicular adenomas), E-cadherin mRNA levels were equally high and the follicles were generally stained, mainly along the lateral surface of the epithelial cells, by the anti-E-cadherin monoclonal antibody. In anaplastic thyroid carcinomas (n = 6) E-cadherin expression was very low or lacking. In papillary carcinomas (n = 23), E-cadherin mRNA levels varied from nearly normal to highly reduced, which roughly correlated with the overall immunofluorescence intensity. However, the immunostaining also revealed a heterogeneous "all-or nothing" expression of E-cadherin among adjacent cells in the same tumor. In the follicular carcinomas (n = 9), E-cadherin mRNA levels were in general rather high but the immunostaining varied considerably. A few papillary and follicular tumors lacked immunoreactive E-cadherin in spite of high mRNA levels. In oxyphilic (Hurthle) cell tumors, comprising both adenomas (n = 4) and carcinomas (n = 2), E cadherin immunoreactivity was reduced and distributed intracellularly rather than at the cell surface. The expression of E-cadherin in relapsing thyroid carcinomas and in tumors with metastatic spreading was, irrespective of the histiotype, low or lacking. Sequential Northern analysis revealed a close correlation between the expression levels of E-cadherin and the thyrotropin receptor. Together, the data suggest that in human thyroid malignancies both gene expression and posttranscriptional control of E-cadherin may be impaired. PMID- 8402690 TI - Mitochondrial alterations in photodynamic therapy-resistant cells. AB - The characterization of radiation-induced fibrosarcoma cells (RIF-8A) which have been selected for resistance to Photofrin-mediated photodynamic therapy (PDT) is detailed in this report. Morphological and functional assessment of mitochondria in both the resistant RIF-8A and parental RIF-1 cells show distinct differences. Electron micrographs show that the mitochondria in the RIF-8A cells are relatively smaller; stain more densely, and display a higher cristae density than RIF-1 cells. P. A. Andrews et al. (Cancer Res., 52: 1895-1901, 1992) reported similar mitochondrial differences between a human ovarian carcinoma cell line, 2008, and its cisplatin-resistant counterpart (C13*). Dose-response curves demonstrate that these cisplatin-resistant C13* cells show cross-resistance to Photofrin-mediated PDT. Functionally, the RIF-8A cells produce more ATP and demonstrate higher succinate dehydrogenase activity than do the RIF-1 cells, but the rates of oxygen consumption do not differ between the two cell types. The PDT sensitive RIF-1 cells demonstrate a significantly higher susceptibility to inhibition of glycolytic activity as determined by 2-deoxy-d-glucose survival curves. These findings suggest differences in the efficacy and/or mode(s) of energy production in the RIF-1 and RIF-8A cells. Since mitochondria are sensitive targets for porphyrin-mediated PDT, the observed changes in mitochondrial structure and/or function may be involved in the PDT resistance seen in RIF-8A cells. PMID- 8402691 TI - Estrogen inhibits the growth of estrogen receptor-negative, but not estrogen receptor-positive, human mammary epithelial cells expressing a recombinant estrogen receptor. AB - Estrogen is essential for the growth of the normal mammary gland and most estrogen receptor (ER)-positive mammary carcinomas. To better understand the differences between the estrogen response pathways in normal and tumor cells, we have stably transfected ER-negative immortal, nontumorigenic human mammary epithelial cells and ER-negative breast cancer cells with an ER-encoding expression vector. Unexpectedly, estrogen treatment (1.0 nM) inhibited the proliferation of ER-transfected nontumorigenic and tumor-derived cells. The control transfectants and parental cells exhibited no response to estrogen concentrations as high as 1.0 microM. This inhibitory effect was attributed to a decreased growth rate and a perturbation of the cell cycle distribution by estrogen treatment of the ER transfectants. The inhibitory response was blocked by cotreatment with the antiestrogen ICI 164,384 as predicted for a pure antagonist of estrogen action. However, treatment with the antiestrogen hydroxytamoxifen caused growth inhibition, implying that hydroxytamoxifen acts as an agonist of estrogen action in ER-transfected cells. Since estrogen is a mitogenic and not a growth-inhibitory stimulus for ER-positive breast cancers and cell lines, we tested the effect of constitutive, high level expression of the ER in ER-positive tumor cells. Stable transfection of ER-positive MCF-7 and T47D cells with the ER expression vector yielded cells with varying amounts of ER. At ER levels comparable to those found in the ER-negative transfected cells, the MCF 7 and T47D ER transfectants were not inhibited by estrogen. These data suggest that ER-positive breast cancer cells can tolerate higher constitutive levels of ER expression than ER-negative cells. The mechanism by which this is accomplished may be an essential step in the process which yields ER-positive tumors. PMID- 8402692 TI - Quantitative comparison between the transplantability of human and murine tumors into the subcutaneous tissue of NCr/Sed-nu/nu nude and severe combined immunodeficient mice. AB - In previous reports, nude mice have demonstrated residual immunoreactivity against xenografts. Severe combined immunodeficient (SCID) mice lack functional T and B-cells. These animals are expected to be better hosts in which to perform preclinical studies on human tumors. The purpose of this study is to quantitate the advantage of SCID mice over nude mice in terms of transplantability of human and murine tumors and the importance of residual immunity in SCID mice. The transplantation assays are described by an assay based on the number of tumor cells required to transplant tumor into 50% of recipients (TD50). Seven human tumors of different histology and four murine tumor cell lines were used. Serial 2-10-fold dilutions of cells were injected (0.1 ml) into the flanks of normal and whole-body irradiated WBI nude and SCID mice. The results showed that in 6 of 6 human tumor cell lines studied, TD50S for SCID mice were 2.4 to 200 times lower than that of nude mice (significant in 5 cell lines). In contrast, in 2 of 3 murine tumors, TD50S in WBI SCID mice were significantly higher than that found in nude mice. When SCID and nude mice received WBI, TD50S were lower than those of nonirradiated animals in 5 of 5 xenografts (significant in 2 cell lines for nude mice and in 5 cell lines for SCID mice). We concluded that WBI SCID mice are significantly better recipients of human tumor xenografts than nude mice. There is a factor of 10-1625 gain in TD50S in favor of the WBI SCID mice when compared to nonirradiated nude mice. WBI has, however, an important effect on SCID mice which may suggest a detectable residual immunoreactivity, perhaps due to natural killer cells. These data demonstrate that WBI SCID mice are better models for human tumor transplantation that nude mice and, although WBI at 6 Gy suppressed significantly the immune system of nude mice, a certain level of immunoreactivity against xenografts is still maintained. PMID- 8402693 TI - Quantitative comparison between the transplantability of human and murine tumors into the brain of NCr/Sed-nu/nu nude and severe combined immunodeficient mice. AB - We have demonstrated (A. Taghian et al., Cancer Res., 53: 5012-5017, 1993) that the take rate of human xenografts in the s.c. tissue of severe combined immunodeficient (SCID) mice is significantly higher than that of nude mice. Earlier, this laboratory reported that the transplantability of tumor xenografts was significantly higher for intracranial (i.c.) injection than for s.c. injection in nude mice. The purpose of this study is to assess: (a) the relative i.c. transplantability of human and murine tumors in comparison with s.c. tissue in SCID mice; (b) the relative i.c. transplantability in SCID mice in comparison to nude mice; and (c) the influence of whole-body irradiation on i.c. transplantability of SCID and nude mice. The assay based on the number of cells required to transplant tumors into 50% of recipients (TD50) was used to describe the transplantability assays. Five human and four murine tumor cell lines were used. Concurrent TD50 assays were performed i.c. in whole-body irradiated and nonirradiated SCID and nude mice. Serial 2-10-fold dilutions of cells were injected in a 10-microliters volume into the right parietal lobe 3 mm below the skin. The results showed that in all tumors studied the i.c. TD50S were significantly lower than the s.c. TD50S by a factor of 1.7-1580. The average enhancement ratio (s.c. TD50/i.c. TD50) in nude mice was twice that in SCID mice. No significant difference was found between the i.c. TD50S in SCID and in nude mice, contrary to the significant difference in s.c. TD50S between both strains of mice (A. Taghian et al., Cancer Res., 53: 5012-5017, 1993). Whole-body irradiation did not significantly affect the i.c. TD50 in nude mice; however, it did affect two of three xenografts in SCID mice. In conclusion, despite the significantly lower s.c. TD50S of human xenografts in SCID mice, i.c. TD50S were almost similar to those of NCr/Sed-nu/nu nude mice. This suggests the presence of different immunoreactivities between nude and SCID mice in s.c. transplantability; however, for i.c. transplantability, nude mice behaved equally as well as SCID mice. The significant enhancement ratio in SCID mice is further evidence that this strain of mice displays a residual systemic immunoreactivity, although the immunoreactivity is significantly lower than that of nude mice. PMID- 8402694 TI - Suppression of T-lymphoma cell apoptosis by monoclonal antibodies raised against cell surface adhesion molecules. AB - We have previously established a mouse malignant T-lymphoma CS-21 cell line that preferentially metastasizes to lymph nodes after s.c. inoculation into BALB/c mice. The CS-21 lymphoma cells were grown in vitro in the presence of CA-12 stromal cells isolated from lymph nodes. When CS-21 cells were cultured alone, they underwent apoptotic cell death with DNA fragmentation. In contrast, the culture of CS-21 cells attached to a monolayer of CA-12 stromal cells prevented CS-21 cell apoptosis and enhanced cell proliferation. To identify the cell adhesion molecules, we developed monoclonal antibodies (mAbs) directed against CS 21 cell surface proteins. Fourteen mAbs partially inhibited the binding of CS-21 cells to a CA-12 stromal cell monolayer. MCS-5 (mAb against CS-21 No. 5) directed against a M(r) 168,000 cell membrane protein and MCS-19 against a M(r) 23,000 protein were found to suppress apoptosis and to stimulate CS-21 cell growth. Soluble factors secreted from CA-12 stromal cells enhanced CS-21 cell growth but were not sufficient to prevent apoptosis. In the presence of both stromal cell secreted factors and mAbs MCS-5 or MCS-19, CS-21 lymphoma cells evaded apoptosis and grew as fast as in the coculture with CA-12 stromal cells. Because of these results, we conclude that CA-12 lymph node stromal cells support CS-21 lymphoma cell growth by secreting paracrine growth factors and by presenting receptors for the M(r) 168,000 and M(r) 23,000 cell surface adhesion molecules of CS-21 cells that transmit signals to prevent CS-21 cell apoptosis. PMID- 8402695 TI - Progressive ovarian carcinoma induces synthesis of type I and type III procollagens in the tumor tissue and peritoneal cavity. AB - Increased serum concentrations of aminoterminal propeptide of type III procollagen occur in advanced ovarian cancer. To study their origin, we compared the expressions of type I and type III procollagens in ovarian tumor tissue and the peritoneal cavity with immunoassays for the propeptide domains of these procollagens. Samples of tumor cyst fluid, peritoneal ascitic fluid, tumor vein blood, and peripheral blood were obtained at operation from 50 women with malignant ovarian neoplasms and 61 women with benign neoplasms. The ascitic fluid concentrations of both type I and type III procollagen antigens were significantly higher in the malignant tumors than in the benign ones, but this difference was evident only for type I procollagen in the tumor cysts. The aminoterminal propeptide of type III procollagen concentration in the peripheral blood was higher in the patients with malignant tumors, whereas the concentrations were similar in the tumor veins. The enhanced type I procollagen synthesis in the malignant tumors did not affect the corresponding antigen in the blood. The findings suggest that progressive ovarian carcinoma invariably induces a fibroproliferative response, characterized by active expression of type I and type III procollagens. The increased circulating aminoterminal propeptide of type III procollagen is derived from the peritoneal cavity rather than from the tumor tissue via the ovarian vein. PMID- 8402696 TI - Regulation of the expression of galactoside-binding lectin during human monocytic differentiation. AB - The widely distributed hL-31 (CBP35, epsilon BP, mL-34, L-29, Mac-2) is a Ca(2+) independent galactoside-binding lectin which functions as a receptor on mammalian cells for glycoproteins containing poly-N-acetyllactosamine side chains. Little is known about the regulation of its expression. The human promyelocytic leukemia cell line, HL-60, was used to determine whether expression of hL-31 (Mac-2) correlated with macrophage/monocyte differentiation. Nondifferentiated HL-60 cells and HL-60 cells grown in the presence of 1.24 microM retinoic acid expressed only trace amounts of hL-31. In contrast, both hL-31 transcripts and protein were detected at 8 h after addition of 17 nM 12-O-tetradecanoylphorbol-13 acetate and reached maximal levels at 24 h. Addition of actinomycin D along with 12-O-tetradecanoylphorbol-13-acetate blocked accumulation of hL-31 mRNA. In contrast, addition of actinomycin D to 12-O-tetradecanoylphorbol-13-acetate treated HL-60 cells that had already accumulated high levels of hL-31 mRNA did not cause significant reduction in RNA levels until 6-8 h had elapsed. Since increased hL-31 expression was not associated with an increase in transcriptional activity of the hL-31 gene, these results suggest that hL-31 expression is regulated at the posttranscriptional level, at least in part, by stabilization of its mRNA. The results also indicate that the processes leading to increased hL-31 expression in HL-60 cells may be specific to differentiation along the monocyte/macrophage pathway. PMID- 8402697 TI - Increased expression of nucleoside diphosphate kinase/nm23 and c-Ha-ras mRNA is associated with spontaneous lung metastasis in rat-transplantable osteosarcomas. AB - The relationship between expression of nucleoside diphosphate kinase (NDP kinase)/nm23, c-Ha-ras, and c-myc genes and metastatic potential was assessed in rat-transplantable osteosarcoma lines, derived from spontaneous and chemical carcinogen (4-hydroxyamino quinoline 1-oxide)-induced osteosarcomas in Fischer 344/NS1c rats. These osteosarcomas possess metastatic potential and highly metastatic lines spontaneous osteosarcoma-selected lung metastatic lesions and 4 hydroxyamino-quinoline 1-oxide-induced osteosarcoma-selected lung metastatic lesions were respectively established by selectively transplanting lung metastatic lesions. Northern blot analysis revealed that the levels of NDP kinase/nm23 and c-Ha-ras gene expression were increased in line with metastatic ability; thus transcript levels were remarkably greater in both spontaneous osteosarcoma-selected lung metastatic lesions and 4-hydroxyamino-quinoline 1 oxide-induced osteosarcoma-selected lung metastatic lesions highly metastatic lines than in their respective low metastatic spontaneous and chemical carcinogen (4-hydroxyamino quinoline 1-oxide)-induced osteosarcoma counterparts. c-myc mRNA expression was observed in all tumor lines, without any correlation with metastatic ability. Southern blot analysis did not show evidence of gross rearrangement or amplification of NDP kinase/nm23, c-Ha-ras, or c-myc genes suggesting regulation of their gene expression at the transcriptional and/or posttranscriptional level. These results indicate that NDP kinase/nm23 and c-Ha ras might be cooperatively involved in a positive manner in signal transduction processes, especially involving G-protein reactions, responsible for metastasis of rat-transplantable osteosarcomas. PMID- 8402698 TI - Molecular mechanisms of tumor development and progression: new targets for prevention, diagnosis, and therapy--a pathology B study section workshop. Working report from the Division of Research grants, National Institutes of Health. PMID- 8402699 TI - The sick child and music. PMID- 8402700 TI - The identification and the education of a pediatric neurosurgeon at an international level. AB - Although the field of application of pediatric neurosurgery is easily identified by the age criterion as well as the specific illnesses of children, the "pediatric neurosurgeon" is still fairly undefined. In fact, the evolution of the process by which the pediatric neurosurgeon is identified is quite different in the various countries, depending on the cultural environment, the socioeconomic level and the age distribution of the population. In North America, in recent years there has been activity aimed at obtaining full independence for pediatric neurosurgery. On the other hand, as a result of inertia, academic tradition and the health care system, Europe only appears to be willing to recognize this subspecialty culturally and to oppose its actual independence from general neurosurgery. PMID- 8402701 TI - Symposium on the education and identification of a pediatric neurosurgeon at an international level. AB - In Japan, pediatric neurosurgery is well covered in the medical schools, and clinical training in this field is included in most programs. However, there are few fulltime pediatric neurosurgeons, and there is no specialty board for pediatric neurosurgery. It is included in the general neurosurgery board examination. PMID- 8402702 TI - Heterogeneity of neurological syndromes in survivors of grade 3 and 4 periventricular haemorrhage. AB - To evaluate the topographical neurological distribution, patterns of abnormal tone and related functional neuromotor impairment after grade 3 and grade 4 intraventricular/periventricular haemorrhage (IPVH), 33 children with previous grade 3 or 4 IPVH of mean gestational age 30.9 weeks (range 25-40 weeks) and mean birth weight 1743 g (range 866-3600 g) were examined neurologically at 4.7 years (range 0.75-10.8 years). Neurological signs were absent in 10/33 cases which were equally distributed between the grade 3 and grade 4 IPVH groups. The largest single topographical neurological distribution was hemiparesis in 8/23, followed jointly by diplegia (cerebral paraplegia) in 6/23 and triplegia in 6/23 cases and finally quadriplegia in 3/23 cases. Grade 4 IPVH tended to result in asymmetrical syndromes, accounting for 7/8 cases of hemiparesis and 5/6 cases of triplegia, whereas all 3/3 cases of quadriplegia followed grade 3 IPVH. The 6/23 cases of diplegia were shared between the grade 3 and grade 4 IPVH groups. Tone was normal in 7/8 of the hemiparetic subjects. Dystonia was the commonest tone abnormality, affecting 8/23 children with neurological disturbance, followed by ataxia/hypotonia in 4/23 and mixed dystonia/hypotonia in 3/23. Only 1/23 cases had signs of spasticity. Spasticity is rare following severe IPVH. Diplegic children had a better functional neuromotor grade than hemiparetic children, who in turn did better than triplegic children. Ataxia hypotonia resulted in better functional outcome than dystronia, which in turn was more favourable than mixed tone patterns. Cranial imaging by ultrasound (US) or computed tomographic (CT) scanning proved an unreliable prognostic indicator except in the case of hemiparesis, for which US scans correctly predicted the affected side in 5/7 cases. The neurological syndromes following severe IPVH differ from the classical encephalopathy of prematurity, and this should lead to a re-appraisal of the trends in the prevalence of cerebral palsy. Caution should be exercised in the interpretation of cranial imaging with regard to pessimistic prognoses in the presence of changes or undue optimism in their absence. PMID- 8402703 TI - Moyamoya disease in childhood: a familial case report. AB - Moyamoya is an obstructive cerebrovascular disease characterized by a cerebral angiographic picture of stenosis or occlusion of main cerebral arteries with an abnormal vascular network at the base of the brain. No definitive cause has been found for this disease and opinion is still divided between a congenital and an acquired etiology. Hemiplegia of sudden onset and epileptic seizures are the prevailing presentation in childhood, while subarachnoid bleeding occurs more frequently in adults. We report a new case of childhood moyamoya with clinical onset of the neurological symptoms within the 3rd year of life; during the child's illness the maternal grandmother presented with moyamoya disease too. Antiaggregating and calcium-antagonist drugs seem effective in preventing further vascular accidents, while a surgical approach is not possible. Computed tomography, single positron emission computed tomography, and magnetic resonance imaging are very useful in the diagnosis of this rare disease. PMID- 8402704 TI - Posterior fossa extradural hematomas in children: report of nine cases. AB - Extradural post-traumatic posterior fossa hematoma is a rare condition estimated to complicate about 0.13% of all craniocerebral injuries, and represents 5.8% of the entire group of extradural hematomas in our records. Nine cases of posterior fossa extradural hematomas are presented. The clinical picture was dominated by headache, vomiting, and gait ataxia. An occipital fracture was seen in 77.7% of the patients. In all cases, the diagnosis was made by computed tomography. The postoperative evolution was good. PMID- 8402705 TI - Injection injury of the sciatic nerve (370 cases). AB - Injury to peripheral nerves due to injections of therapeutic and other agents is common. The postulated mechanisms of injury include direct needle trauma, secondary constriction by scar, and direct nerve fiber damage by neurotoxic chemicals in the injected agent. Neurological sequelae can range from minor transient sensory disturbance to severe sensory disturbance and paralysis with poor recovery. The recommended treatment has ranged from a conservative approach to immediate operative exposure and irrigation, and has also included early neurolysis or delayed exploration with neurolysis or resection and anastomosis. We present 370 cases of injection injury of the sciatic nerve in children treated during the last 20 years at the Neurosurgical Department of the Hospital La Paz in Madrid, Spain. Pathology, clinical course, treatment, and results are discussed. PMID- 8402706 TI - Childhood intervertebral disc calcification. AB - Two cases of intervertebral disc calcification in children are reported. A 13 year-old boy presented with right subscapular pain radiating into the axilla with radiographic demonstration of multiple calcified intervertebral discs and a herniated fragment of calcified nucleus pulposus at T2-3. His condition improved with conservative therapy, and follow-up radiographic evaluation revealed resolution of the herniated calcified disc material. A second case of multiple intervertebral disc calcifications was observed incidentally in an asymptomatic 6 year-old boy. The clinical presentation of this condition, its incidence, natural history, proposed etiologies, treatment options, and outcome are reviewed. PMID- 8402707 TI - Pediatric Creutzfeldt-Jakob disease: probable transmission by a dural graft. AB - A 10-year-old boy underwent a posterior fossa craniectomy for removal of a grade 2 cerebellar astrocytoma. Dural closure was achieved by the placement of a dural graft. Eight years later the patient developed dementia and myoclonus. Electroencephalography demonstrated generalized slow activity that evolved into a pattern of periodic triphasic waves. Computed tomography scan and magnetic resonance imaging were unremarkable. Brain biopsy confirmed spongiform encephalopathy of the Creutzfeldt-Jakob type. In the light of previous reports of four similar occurrences, and of our own experience with two further cases of this disease, we believe that the cadaveric dura was the source of transmission of Creutzfeld-Jakob disease in our patient. The authors remark the importance of the awareness of this late complication of dural substitutes, both for the diagnosis of possible future cases and for taking preventive measures to stop the spread of the disease. PMID- 8402708 TI - Development of akinetic mutism and hyperphagia after left thalamic and right hypothalamic lesions. AB - A case of childhood post-traumatic akinetic mutism is presented. The patient showed a hyperphagic condition while recovering from akinetic mutism. He had lesions in the left interlaminal nucleus of the thalamus, right globus pallidus, and right dorsomedial nucleus of the hypothalamus. Laboratory data indicated slightly disturbed hypothalamic functions. In general, akinetic mutism can be seen with bilateral destructive lesions, while hyperphagia may occur after destruction of dorsomedial hypothalamic nucleus, but it is very rare. This is the first reported case of akinetic mutism caused by a unilateral lesion. PMID- 8402709 TI - Intracranial solitary-type infantile myofibromatosis. AB - An unusual case of infantile myofibromatosis of the solitary type occurring in an intracranial location in a 48-day-old female infant is presented. To our knowledge, there are no other descriptions in the literature of infantile myofibromatosis with exclusively intracranial involvement. The immunohistochemical and electron microscopic findings confirm the myofibroblastic origin of the proliferation. PMID- 8402710 TI - Spinal extradural arachnoid cyst. AB - The case of 16-year-old boy with a spinal extradural arachnoid cyst is presented. An extradural arachnoid diverticulum extending from T10 to L1 was excised totally with hemilaminectomy. Surgery caused prompt improvement of the neurological deficit. The pertinent literature is reviewed. PMID- 8402711 TI - Molecular implications of recurrent cytogenetic alterations in human small cell lung cancer. AB - This article describes cytogenetic findings in human small cell lung cancer (SCLC). Included is a summary of analyses performed by the authors on 17 tumors, each of which displayed numerous chromosomal alterations. Many of the recurrent changes involve losses at the locations of tumor suppressor genes, whose loss and/or inactivation may play a crucial role in tumorigenesis. Deletions of the short arm of chromosome 3 (particularly 3p21-25) were found in every case, providing additional evidence in support of the notion that this region harbors a tumor suppressor gene(s) critical in the pathogenesis of SCLC. Cytogenetic losses of 5q21, 13q14, and 17p13 (sites of the APC, RB1, and TP53 suppressor loci, respectively) also are common in SCLC. Double minutes are found in a minority of these tumors and are associated with oncogene amplification. The genetic complexity in SCLC underscores the need for greater preventive measures and early detection. PMID- 8402712 TI - APC gene mutations in Italian familial polyposis coli patients. AB - Adenomatous polyposis coli is an autosomal dominant disease characterized by the development of hundreds of colorectal adenomas in young adults. If prophylactic colectomy is not performed, colorectal cancer develops in virtually all affected individuals by the fifth decade of life. All at-risk relatives older than 10 years of age need to be screened regularly by endoscopy. Recently, the gene responsible for the disease, the APC gene, was cloned. The finding of inactivating mutations of the APC gene in Italian APC patients allowed us to offer DNA-based diagnostic tests to these families. PMID- 8402713 TI - Automated DNA sequencing: a look into the future. PMID- 8402714 TI - Recombinant immunotoxins: new therapeutic agents for cancer treatment. AB - We have developed new agents for the treatment of cancer by genetically modifying Pseudomonas exotoxin. We have deleted the cell-binding region of Pseudomonas exotoxin and replaced it with various growth factors or the combining regions of antibodies in a single chain form. These new recombinant molecules are called recombinant toxins. Several different types of recombinant toxins have been produced. B3(Fv)-PE38KDEL is a recombinant toxin that kills many different adenocarcinomas and epidermoid carcinomas. The molecule is now undergoing preclinical development. PMID- 8402715 TI - Use of anti-CD3 and anti-CD16 bispecific monoclonal antibodies for the targeting of T and NK cells against tumor cells. AB - To target T lymphocytes against EGF-R+ tumors, we constructed anti-CD3/anti-EGF-R bimAbs either by the generation of a hybrid hybridoma (quadroma) or by a chemical cross-linking method. Analysis of the in vitro functional activity of these two different constructs indicated that the quadroma-secreted bimAb was more efficient in targeting the CD3+8+ clones against EGF-R+ target cells with respect to the bimAb produced by chemical method. In addition, the quadroma-produced bimAb is able to induce cytolysis of EGF-R+ tumor cell lines of PHA-induced lymphoblasts that had been expanded in IL-2-containing medium, whereas tumor cells lacking expression of EGF-R were not lysed. Resting PBL targeted by the bimAb did not display significant cytotoxicity against the relevant tumor. An anti-CD16 hybridoma (IgG1) was fused with an anti-folate-binding protein hybrid (IgG2a) to construct bimAbs to target NK cells against NK-resistant ovarian carcinomas. The hybrid IgG1/IgG2a bimAb triggered the specific lysis of relevant target cells by resting NK cells, but it was ineffective when CD8+TCR alpha/beta+ cultured cell populations were used as effectors. Only marginal increases of cytolytic activity could be induced by the bimAb when IL-2-activated PBL (i.e., LAK cells) were used as effectors due to the high cytolytic activity of these cells against the relevant tumors in the absence of bimAb. The possible use of anti-CD16 or anti-CD3 bimAbs for the development of different cellular immunotherapy strategies against cancer is discussed. PMID- 8402716 TI - The production of human lactoferrin in the milk of transgenic animals. PMID- 8402717 TI - Production of human hemoglobin in transgenic swine: an approach to a blood substitute. AB - Many significant challenges have to be met before a clinically useful hemoglobin based blood substitute can be developed. A major issue is the choice of a production system. Transgenic swine have several features that appear to be suited to the abundant production of human hemoglobin. Here we show that authentic human hemoglobin can be made in transgenic pigs and purified by simple ion-exchange chromatography. The purified hemoglobin can be modified for testing and use as a blood substitute. PMID- 8402718 TI - Biotechnological patents and ethical aspects. AB - Developments in the biotechnological fields have stimulated public research institutes to adopt a more enterprising policy that encourages the use of the patent and the commercialization of research results and products. The National Institute for Cancer Research of Genoa (IST) has dealt with a number of the principles and ethically related matters that arise from the acceptance of the patent system and policy. The problem of the patent in the biotechnological fields is particularly complicated given the subject--living matter--and the social alarm that it provokes in the general public. Careful consideration of ethical aspects demands the further evaluation of whether or not the patent is an adequate tool for regulating these matters. PMID- 8402719 TI - Biotechnology patents and ethical aspects. AB - Should we set any limits on patenting? More specifically, must we set any limits on patenting in the field of biotechnology? There should be general agreement on the exclusion of humans from patentability. The European Parliament voted unanimously on an amendment to the Community Directive regarding patenting stating that the human body and its parts are not patentable as such. Patenting of humans indeed would be against fundamental human rights; against the shared principles of freedom, autonomy, and dignity of each single human being. The same reasons apply to requests to reject the "commercialization of the human body." However, much more difficult is reaching a consensus on what are the parts of humans that should not be marketed--organs, tissues, cells, genes, smaller DNA fragments? Probably there is no consensus on where to draw the line when we deal with parts of the human body. Nevertheless, an ethical component is very strong in raising opposition to patenting human DNA. Whatever our personal view on the issue, we cannot deny that ethical aspects must be considered in granting patents on human DNA. With reference to animals, the fears raised are that the patenting of transgenic animals could amplify the instrumental use (reification) and the neglect of their sentient nonobjectual nature: patenting could motivate, instead, the tendency to consider animals as the standard of things invented and as new consumer products. Moreover, animal patenting increases production and thus brings about the great suffering of animals. In regard to plants, the ethical implications of patenting have more to do with their socioeconomic effects, in particular on Third World countries, than for the organisms involved.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402720 TI - Ethical aspects of patenting in biotechnology. PMID- 8402721 TI - Biotechnology and developing countries. PMID- 8402722 TI - Recruiting older women for screening mammography. AB - We compared health behavior and attitudes of older and younger women toward breast cancer screening by using data from the 1987 Texas Breast Screening Project, a community-based, low-cost screening program sponsored by the American Cancer Society, Texas Division. Because age is an important risk factor for breast cancer, the women were categorized into three age groups: 55 to 64 years old, 65 to 74 years old, and 75 years and older. Approximately 67% of the women 75 years and older, 64% of women in the 65 to 74 age group, and 59% of the younger (55 to 64) women had never had mammography. Moreover, only 14% of the women in both the 65 to 74 and 75+ age groups and 19% of the younger women reported having two or more mammographic examinations. Fewer older women (65 to 74 and 75+ age groups) reported having recent clinical breast examination. There were no differences among the age groups in the factors that attracted the women to participate in this screening program; in the three age groups, the most influential factors to participate were media publicity and lower mammography costs. Women in all age groups reported a lack of physician referral and cost as reasons for not previously participating in mammography screening. This study shows that older women are underscreened. Educational programs about the benefits of early detection should target older women because they are at increased risk for breast cancer. PMID- 8402723 TI - Barriers to screening among participants of a media-promoted breast cancer screening project. AB - Breast cancer detection and awareness projects have been implemented nationwide in an attempt to increase compliance with screening mammography. Previous studies, however, showed that the elderly, minorities, and women of lower socioeconomic status fail to respond in representative numbers. A cross-sectional analysis of 6640 participants of a Breast Cancer Detection and Awareness Project in Tampa, FL, was conducted to determine if barriers and motivations to screening differed among targeted (the elderly, minorities, women of lower socioeconomic status) and nontargeted groups. Targeted demographic groups reported far more barriers to screening and fewer motivating factors in their decision to participate in screening. This was true for the elderly, minorities, and women of lower socioeconomic status. Women in greater need of screening mammography report fewer motivations, and must overcome greater barriers to participate in media promoted breast screening projects. Changes in the design and promotion of these screening projects must occur to prevent reverse targeting. PMID- 8402724 TI - Relationship between immunohistochemically detectable p53 protein and prognostic factors in head and neck tumors. AB - This study examined the relationship between immunohistochemically detectable p53 protein and prognostic factors in squamous cell carcinoma of the head and neck (SCCHN). Twenty-seven tumor specimens were evaluated utilizing a panel of three monoclonal antibodies (MAbs) directed against different epitopes of the p53 protein (PAb 421, PAb 1801, and PAb 240). The overall incidence of p53 protein detection with a panel of MAbs was 78%, which was significantly higher than with any one of the tested antibodies. Comparison of the tumors that were negative for p53 with tumors that stained positive with one or multiple antibodies, however, revealed no statistically significant differences with respect to the stage of disease, metastatic node involvement, size of the primary tumor, or degree of tumor differentiation. The results of our study suggest that levels of p53 protein, although commonly immunohistochemically detected in head and neck tumors, do not correlate with known prognostic factors for SCCHN. PMID- 8402725 TI - Cathepsin B and cysteine proteinase inhibitors in bronchoalveolar lavage fluid of lung cancer patients. AB - The enzymatic activity and the concentration of cathepsin B (CB), determined by ELISA, and total inhibitory activity of cysteine proteinase inhibitors (CPIs) were measured in bronchoalveolar lavage fluid (BALF) of lung tumor patients (n = 49). Significantly higher CB activity and concentration was found in BALF from metastasis (n = 15), when compared to squamous cell carcinoma (SCC; n = 15) and small cell lung carcinoma (SCLC; n = 7). Patients with adenocarcinoma (n = 12) also secreted considerably more CB, about 14- and 3.3-fold, compared to SCC and SCLC patients, respectively. However, only tumor patients without inflammation were considered because the comparison of patients with lung tumors (n = 49) and with other non-neoplastic lung diseases (n = 18) showed no differences due to high CB in BALF of patients with inflammatory conditions, present in some patients from both groups. Immunolabeling of tumor BALF proteins revealed CB immunoreactivity at molecular sizes that corresponded to the size of its precursor and mature forms, as well as to complexes with kininogen(s) and low molecular weight CPIs. Stefins A and B, but not cystatins C and S appeared in the complexes with CB. PMID- 8402726 TI - Genomic analysis of human hepatocellular carcinomas using Restriction Landmark Genomic Scanning. AB - Restriction Landmark Genomic Scanning (RLGS) was used to examine the multiple alterations of genomic DNAs that occur in association with transformation and development of malignancy in primary hepatocellular carcinoma (HCC). Genomic DNAs from HCC and its normal counterpart were cleaved by the restriction enzyme NotI, radiolabeled at the cleavage sites, and then size-fractionated by two-dimensional electrophoresis using HinfI as the second cleavage enzyme. About 2000 spots were recognized, whose position and intensity reflect the locus and the copy number of the corresponding restriction sites. Using this system in combination with micromanipulation of HCC to eliminate possible carry-over of nonmalignant cells, we detected six spots that were decreased in intensity in common to three different HCCs, along with five that were intensified spots. In addition, several spots showed changes that were nonoverlapping among different tumors. PMID- 8402727 TI - The putative tumor suppressor gene on chromosome 5q for hepatocellular carcinoma is distinct from the MCC and APC genes. AB - We have previously shown that the tumor suppressor gene for hepatocellular carcinoma (HCC) without cirrhosis may be located on chromosome 5q35-qter. In this study, we analyzed nine cases of primary HCC without cirrhosis using probes from the MCC and APC genes, which are in the region 5q21-22. None of the informative cases had allele loss detected by these probes, whereas the probe lambda MS8 for the region 5q35-qter showed allele loss in six out of six informative cases. The results confirm that the putative tumor suppressor gene for HCC without cirrhosis on chromosome 5q is distinct from the MCC and APC genes. PMID- 8402728 TI - Serum CEA, CA 15-3, and MCA in breast cancer patients: a clinical evaluation. AB - With the aim of investigating the clinical usefulness of CEA, CA 15-3, and MCA serum levels, we studied 143 women whose breast cancer was submitted to serial tumor marker determinations: 79 women had stage I-II tumors and were undergoing follow-up after local and adjuvant treatment; 64 women presented metastatic lesions. Among the stage I-II patients, 63 women remained disease-free during the observation period and 16 developed metastases. In 13 out of 16 patients, tumor markers were elevated and in 11 out of 16 patients the increased tumor markers were the first sign of disease progression. Among metastatic patients, 49 presented increased tumor markers and 15 normal value. Moreover, we observed a decrease or normalization of tumor markers in patients responding to treatment and increased values in progressive disease. No correlation was noted between site of disease and tumor markers. We concluded that tumor markers are of clinical value in the detection of metastasis and may be useful in monitoring response to treatment in metastatic patients. PMID- 8402729 TI - The value of follow-up after curative surgery of colorectal carcinoma. AB - Between 1978 and 1989, 1045 of 1399 patients (580 male and 474 female) had undergone curative surgery for colorectal carcinoma. Of these patients, 350 (33%) had recurrences, another 16 (1.5%) developed a metachronous colorectal cancer, and 23 (2%) had cancers of other organs. An isolated locoregional recurrence was found in 75 of 350 (21%). The remaining 275 of 350 (79%) of the patients showed systemic dissemination of the carcinoma. Reoperations with curative intent were performed on 56 of 350 (16%) of the patients. Only 21 of the 56 resected patients (38%), i.e., 21 of 350 (6%), are without recurrence at the end of the follow-up period on December 31, 1990. Despite a curative reoperation, 62% of the patients again developed recurrent growths. There is an imbalance of the efforts invested in tumor follow-up and the benefits gained. Further follow-up programs should be investigated in a controlled, prospective fashion. PMID- 8402730 TI - Doxorubicin, vincristine, and actinomycin-D, but not teniposide, require long lasting uninterrupted verapamil pressure to overcome drug resistance in multidrug resistant cells. AB - The cytotoxic efficacy of doxorubicin (DOX), vincristine (VCR), actinomycin-D (ACT-D), and teniposide (VM-26) toward the LoVo and SW948 multidrug-resistant (MDR) cell sublines was significantly enhanced by the concomitant presence of verapamil (VER) during pharmacological treatment (1-h exposure) without, however, achieving a complete reversion of the MDR phenotype. Long-lasting VER in the culture medium, at the end of the combined drug+VER treatment, conferred to DOX, VCR, and ACT-D cytotoxic efficacy roughly similar to that exerted on the drug sensitive parent cell lines. On the contrary, no enhancement of VM-26 cytotoxic efficacy was derived from continuous VER treatment. Enhancement of DOX, VCR, and ACT-D cytotoxic efficacy requires that VER be present in an uninterrupted manner in the culture medium since brief interruptions (15 to 60 min) in the VER pressure completely counteract any effect. These findings offer new insight into the modalities of pharmacological treatment that MDR cells require to obtain a complete reversion of cellular resistance to the various drug families included in the MDR spectrum. PMID- 8402731 TI - Scintigraphic detection of xenografted tumors producing human basic fibroblast growth factor. AB - A murine monoclonal antibody 3H3 recognizes the basic fibroblast growth factor (FGF) and inhibits the growth of human glioblastoma cells both in vitro and in vivo. We studied the potential of a scintigraphic technique using the 3H3 antibody to detect tumors that produce basic FGF. 125I- and 111In-labeled 3H3 bound to U87MG human glioblastoma cells in vitro. U87MG cells were inoculated subcutaneously into nude mice. After development of the tumor, radiolabeled 3H3 was injected into the subcutaneous space surrounding the tumor. A high level of radioactivity from 3H3 was retained at the tumor, whereas an irrelevant antibody cleared rapidly from the injected site. Radiolabeled 3H3 was not retained in tumors that did not produce basic FGF. Scintigraphic detection of tumors expressing basic FGF would be valuable for the therapeutic application of the antibody. PMID- 8402732 TI - Retroviral transduction of interferon-gamma cDNA into a nonimmunogenic murine fibrosarcoma: generation of T cells in draining lymph nodes capable of treating established parental metastatic tumor. AB - Gene modification of tumor cells with the cDNA for interferon gamma (IFN gamma) has been shown to increase the immunogenicity of some tumor cells. In order to explore further the possible therapeutic relevance of these previous findings, two clones of the nonimmunogenic MCA-102 fibrosarcoma of C57BL/6 origin were retrovirally transduced with the cDNA encoding murine IFN gamma: 102.4JK (4JK), a clone with relatively high major histocompatibility complex (MHC) class I expression, and 102.24JK (24JK), a clone with low expression of surface MHC class I molecules. Retroviral transduction of tumor cells with the cDNA encoding for IFN gamma resulted in a substantial up-regulation of MHC class I surface expression in the 24JK clone but little change of class I in the 4JK clone. In an attempt to generate antitumor lymphocytes, these gene-modified cells were inoculated into mouse footpads and draining lymph nodes (DLN) were removed, dispersed, and cultured in vitro for 10 days with irradiated tumor cells and interleukin-2. DLN from mice bearing either unmodified tumor or tumor transduced with cDNA encoding neomycin resistance (NeoR) or IFN gamma, were used to treat recipients harboring 3-day pulmonary metastases induced by the parental, unmodified tumor. Treatment with DLN cells obtained following the injection of 24JK tumor cells modified with the gene for IFN gamma significantly reduced the number of pulmonary metastases in four separate experiments, compared to groups treated by DLN cells generated from inoculation of either the unmodified, parental 24JK clone or the same clone transduced with the NeoR gene only. In contrast, DLN cells induced either by IFN gamma-transduced 4JK (high expression of MHC class I) or an unmodified 4JK tumor (moderate expression of MHC class I) had significant but equal therapeutic efficacy. Although the in vitro growth rate of tumor cell lines was unaffected by the insertion of the mouse IFN gamma cDNA, their in vivo (s.c.) growth rates were significantly slower than those of the nontransduced tumors. Thus, after retroviral transduction of the murine IFN gamma cDNA into a nonimmunogenic tumor with a very low level of surface expression of MHC class I, modified tumor cells could elicit therapeutic T cells from DLN capable of successfully treating established pulmonary metastases upon adoptive transfer. This strategy significantly confirms previous observations on the potential therapeutic effects of gene modification of tumor cells with IFN gamma and extends the realm of therapeutic possibilities to include the use of DLN cells for the development of T-cell based immunotherapies against nonimmunogenic human tumors. PMID- 8402733 TI - Antitumor response to recombinant murine interferon gamma correlates with enhanced immune function of organ-associated, but not recirculating cytolytic T lymphocytes and macrophages. AB - The mechanism of therapeutic activity for recombinant murine interferon-gamma (rMu IFN gamma) in the treatment of metastatic disease was investigated by comparing effector cell augmentation with therapeutic activity in mice bearing experimental lung metastases (B16-BL6 melanoma). Effector cell functions in spleen, peripheral blood, and lung (the tumor-bearing organ) were tested after 1 week and 3 weeks of rMu IFN gamma administration (i.v. three times per week). Natural killer (NK), lymphokine-activated killer (LAK), cytolytic T lymphocyte (CTL) activities against specific and nonspecific targets, and macrophage tumoristatic activity were measured. rMu IFN gamma demonstrated immunomodulatory activity in most assays of immune function. The optimal therapeutic protocol of rMu IFN gamma (2.5 x 10(6) U/kg, three times per week) prolonged survival and decreased the number of pulmonary metastatic foci. This therapeutic activity was correlated with specific CTL activity from pulmonary parenchymal mononuclear cells (PPMC), but not from spleen or blood. Macrophage tumoristatic activity in PPMC also correlated with therapeutic activity, but activity in alveolar macrophages did not. However, therapeutic activity did not correlate with NK or LAK activity at any site. These results demonstrate that the optimal therapeutic protocol is the same as the optimal immunomodulatory dose for pulmonary CTL and macrophage activities. Furthermore, while immunological monitoring may help to optimize treatment protocols, current monitoring procedures that use readily accessible sites, particularly peripheral blood, may not accurately predict the therapeutic efficacy of biological response modifiers in clinical trials. PMID- 8402734 TI - Immunomodulatory effects of ultra-low-dose interleukin-2 in cancer patients: a phase-IB study. AB - High-dose interleukin-(IL-2) has been broadly studied in tumor therapy, yet it may be inhibitory to T-cell-dependent immunity. Therefore immune and tumour responses mediated by low-dose IL-2 were studied systematically with respect to the feedback organisation of immune responses. IL-2 was administered once daily at three dose levels: 0.18, 0.9, 4.5 MIU/m2 according to three different schedules requiring subcutaneous (s.c.) injection once weekly (four doses, stratum I), thrice weekly every other day (nine doses, stratum II), or five times weekly every other week (ten doses, stratum III). A total of 46 patients with advanced cancer were randomly assigned to one of the nine treatment groups. Systemic effects were induced at doses as low as 0.18 MIU/m2 IL-2 s.c. as demonstrated from measurable IL-2 serum levels, induction of circulating IL-6, a transient lymphopenia, and stimulation of delayed-type hypersensitivity (DTH) responses of the skin. Analysis of the different IL-2 schedules demonstrated (a) prolonged effects of once-weekly injections on DTH responses, lymphocyte and eosinophil counts, and (b) maximum increase of eosinophil counts and preferential expansion of activated NK cells with repeated injections every 48 h or 72 h (stratum II), while sequential treatment according to stratum III was found to be more potent in increasing the number of activated T cells. A tumour response was observed in 1/15 patients with renal cell carcinoma who experienced more than 50% tumour regression for 8 months; 12 patients had stable disease for 4 months (median). These data demonstrate prolonged immunological effects of ultra-low doses of s.c. IL-2 despite its short half-life. Furthermore, scheduling of IL-2 was found to affect immune responsiveness specifically as demonstrated by the differential effects on natural killer and T cell populations. PMID- 8402735 TI - A protein fraction from aged garlic extract enhances cytotoxicity and proliferation of human lymphocytes mediated by interleukin-2 and concanavalin A. AB - Fraction 4 (F4), a protein fraction isolated from aged garlic extract, enhanced cytotoxicity of human peripheral blood lymphocytes (PBL) against both natural killer (NK)-sensitive K562 and NK-resistant M14 cell lines. Although F4 treatment alone increased cytotoxicity, the effect was more remarkable when F4 was administered together with suboptimal doses of interleukin-2 (IL-2); combination treatment of 5 micrograms/ml F4 plus 10 U/ml IL-2 for 72 h generated lymphokine activated killer activity equivalent to that produced by 100 U/ml IL-2 alone against M14. F4 enhanced IL-2-induced proliferation and IL-2 receptor (Tac) expression of PBL without significant increase of IL-2 production. The enhancement of cytotoxicity both by F4 alone and by F4 plus IL-2 was abolished by anti-IL-2 antibody. F4 also enhanced concanavalin-A(ConA)-induced proliferation of PBL. Radiolabeled-ConA binding assays revealed that F4 treatment greatly augmented the affinity and slightly increased the number of ConA binding sites in PBL. F4 also enhanced ConA-induced IL-2 receptor (Tac) expression and IL-2 production of PBL. Anti- IL-2 antibody inhibited the effect of F4 on ConA-induced proliferation. These data suggest that IL-2 is involved in augmentative effects of F4. Our results indicate that F4 is a very efficient immunopotentiator and may be used for immunotherapy. PMID- 8402736 TI - Efficient expansion of tumor-infiltrating lymphocytes from solid tumors by stimulation with combined CD3 and CD28 monoclonal antibodies. AB - Combined CD3 and CD28 monoclonal antibodies (mAb) may initiate efficient activation and expansion of tumor-infiltrating lymphocytes (TIL). In this study we compared phenotypical and functional characteristics of TIL from a group of 17 solid human tumors, stimulated either by high-dose recombinant interleukin 2 (rIL 2, 1000 IU/ml) or by a combination of anti-CD3 and anti-CD28 monoclonal antibodies in the presence of low-dose rIL-2 (10 IU/ml). Compared to activation with high-dose rIL-2, stimulation of TIL with CD3/CD28 mAb induced significantly stronger proliferation and yielded higher levels of cell recovery on day 14. Following the CD3/CD28 protocol, expansion of an almost pure population of CD3+ cells was obtained. Whereas CD4+ cells dominated in the first week of culturing, within 4 weeks the CD8+ population increased to over 90%. The specific capacity to kill autologous tumor cells was not increased as compared to the high-dose rIL 2 protocol, but all cultures showed high cytotoxic T cell activity as measured in a CD3-mAb-mediated redirected kill assay. These studies show that combined CD3 and CD28 mAb are superior to rIL-2 with respect to the initiation of expansion of CD8+ cytolytic TIL from solid tumors. Stimulation with specific tumor antigens at a later stage of culturing may further augment the expansion of tumor-specific cytolytic T cells. PMID- 8402737 TI - Drug resistance in rat colon cancer cell lines is associated with minor changes in susceptibility to cytotoxic cells. AB - The development of resistance to anticancer drugs urges the search for different treatment modalities. Several investigators have reported the concomitant development of drug resistance and resistance to natural killer (NK), lymphokine activated killer (LAK) or monocyte/macrophage cell lysis, while others described unchanged or even increased susceptibility. We investigated this subject in the rat colon carcinoma cell line, CC531-PAR, which is intrinsically multidrug resistant (MDR), and in three sublines derived from this parental cell line: a cell line with an increased MDR phenotype (CC531-COL), a revertant line from CC531-COL (CC531-REV), which demonstrates enhanced sensitivity to anticancer drugs of the MDR phenotype, and an independently developed cisplatin-resistant line (CC531-CIS). In a 4-h 51Cr-release assay we found no difference in susceptibility to NK cell lysis. No significant differences in lysability by adherent LAK (aLAK) cells were observed in a 4-h assay. In a prolonged 20-h 51Cr release assay an enhanced sensitivity to aLAK-cell-mediated lysis was observed in the revertant, P-glycoprotein-negative cell line and in the cisplatin-resistant cell line (CC531-CIS). None of the cell lines was completely resistant to lysis by aLAK cells. Therefore, a role for immunotherapy in the treatment of drug resistant tumors remains a realistic option. PMID- 8402738 TI - Therapeutic potential of chimeric and murine anti-(epidermal growth factor receptor) antibodies in a metastasis model for human melanoma. AB - On many tumors, high numbers of epidermal growth factor (EGF) receptors provide a target for antibody-mediated tumor therapy. We evaluate here the therapeutic potential of a mouse/human chimeric anti-(EGF receptor) antibody and compare it to the parental murine antibody in a xenograft model for metastatic melanoma. Our model is based on the human cell line M24met, which overexpresses the EGF receptor and metastasizes spontaneously in SCID mice. Both the chimeric anti-(EGF receptor) antibody (ch225) and the mouse monoclonal antibody (m225) exhibited saturable, high-affinity binding to M24met cells and were equivalent in their ability to target M24met tumors in mice. Neither anti-(EGF receptor) antibodies nor EGF modulated the growth of M24met cells in vitro. Further analysis revealed that the EGF receptor on these cells is not phosphorylated upon EGF binding, indicating an anomalous receptor on these cells. In antibody-dependent cellular cytotoxicity experiments, ch225 and m225 were potent mediators of M24met cytolysis by effector cells. Antibody-mediated cytotoxicity revealed a marked species preference, with ch225 activating human peripheral blood mononuclear cells and m225 activating mouse splenocytes and to a lesser degree mouse macrophages. Neither antibody mediated cytolysis in the presence of human complement. In SCID mice, m225 suppressed spontaneous metastasis considerably while ch225 had only a modest effect. Our data indicate that in the M24met melanoma tumor model, anti-(EGF receptor) antibodies suppress spontaneous metastasis solely by activating immune effector cells. PMID- 8402739 TI - [The effect of maximal vasodilation on the distensibility of the coronary vascular bed]. AB - This study aimed at investigating the changes in coronary vascular resistance induced by sudden increases in transmural pressure in the presence of a maximally vasodilated coronary bed. In anaesthetized open-chest dogs under artificial ventilation, aortic blood pressure, left ventricular pressure and the flow in the left circumflex coronary artery were recorded. The flow was derived by means of an electromagnetic flowmeter. Maximal vasodilatation was achieved by intracoronary infusion of dipyridamole (10-40 mg/h), increases in transmural pressure, starting from 70 mmHg, were obtained by constricting for 10 s the descending thoracic aorta with a plastic snare. While in the absence of vasodilatation the increase in pressure was accompanied with an increase in resistance because of an auto-regulatory response, when the coronary bed was maximally dilated the increase in pressure did not alter the coronary vascular resistance. These results seem to be in conflict with the observation that in the coronary circulation the distension of the vascular wall produced by increases in pressure is favoured by the reduction of the vasomotor tone. However, it may be argued that, while a reduction of the vasomotor tone can increase the vascular distensibility, a maximal vasodilatation, as it was seen in the resistance vessels of the skeletal muscles, brings the vascular diameter to such a size that no further distension can be induced when the transmural pressure is increased starting from a value of about 70 mmHg. PMID- 8402740 TI - [The efficacy of slow-release diltiazem in the treatment of stable angina of effort: a comparison between diltiazem and placebo]. AB - The efficacy of a new slow release (SR) diltiazem preparation was assessed in 10 patients with stable effort angina. A double-blind, placebo controlled, randomized, crossover protocol was carried out comparing the effects of diltiazem 60 mg tid and diltiazem SR 120 mg bid on clinical and ergometric parameters. Exercise test was carried out 3 and 12 hours after the last dose of diltiazem or diltiazem SR respectively. Both drug preparations reduced the incidence of positive test, increased the exercise time and the time of onset of ischemic ST depression. The beneficial effects of the drugs appeared to be due to a reduction in myocardial oxygen consumption at the same workload as shown by the lesser value of pressure rate product at submaximal exercise. In conclusion, diltiazem SR at 12 hours after the last administration has the same effectiveness of diltiazem 60 mg at 3 hours. PMID- 8402741 TI - [Cutaneous microcirculation and diabetic disease. A functional and flowmetry study in subjects with diabetes mellitus type 2]. AB - Laser-Doppler single fingertip skin blood flow has been evaluated in 41 euglycemic type II diabetic patients under basal conditions and after dynamic testing (both ischemia and thermal stress). The same subjects have also undergone tests for the assessment of the degree of autonomic nervous system (ANS) dysfunction. The results have been compared to those obtained in 38 age-matched healthy subjects. In diabetic patients: baseline flow levels were much higher; the post-ischemic flow increase was less evident; a shorter hyperemic phase followed ischemia; a longer latency period was noticed, during thermal stress, together with a lower and slower hyperemic peak level. According to the results of ANS dynamic tests, diabetic subjects were divided into 3 groups: Group 1 (subjects with negative results); Group 2 (subjects with only one positive result); Group 3 (subjects with more than one positive test). Microcirculation disturbances were more often found in Group 3. These results show that a correlation exists between diabetic microangiopathy and ANS dysfunction. They also support the hypothesis, already pointed out by other research groups, of a similar mechanism causing diabetic neurologic and vascular complications. PMID- 8402742 TI - [The development of a computer model in the quantitative assessment of thallium 201 myocardial scintigraphy]. AB - Thallium-201 scintigraphy is a widely used noninvasive procedure for the detection and prognostic assessment of patients with suspected or proven coronary artery disease. Thallium uptake can be evaluated by a visual analysis or by a quantitative interpretation. Quantitative scintigraphy enhances disease detection in individual coronary arteries, provides a more precise estimate of the amount of ischemic myocardium, distinguishing scar from hypoperfused tissue. Due to the great deal of data, analysis, interpretation and comparison of thallium uptake can be very complex. We designed a computer-based system for the interpretation of quantitative thallium-201 scintigraphy data uptake. We used a database (DataEase 4.2-DataEase Italia). Our software has the following functions: data storage; calculation; conversion of numerical data into different definitions classifying myocardial perfusion; uptake data comparison; automatic conclusion; comparison of different scintigrams for the same patient. Our software is made up by 4 sections: numeric analysis, descriptive analysis, automatic conclusion, clinical remarks. We introduced in the computer system appropriate information, "logical paths", that use the "IF ... THEN" rules. The software executes these rules in order to analyze the myocardial regions in the 3 phases of scintigraphic analysis (stress, redistribution, re-injection), in the 3 projections (LAO 45 degrees, LAT,ANT), considering our uptake cutoff, obtaining, finally, the automatic conclusions. For these reasons, our computer-based system could be considered a real "expert system". PMID- 8402743 TI - [Aortic dissection without intimal laceration: a case report and review of the problem]. AB - A 43-year-old hypertensive woman with symptoms of aortic dissection was referred to our hospital 3 days after the initial episode. Transesophageal echocardiography and magnetic resonance showed a large intramural hematoma of the ascending and descending aorta without intimal flap and tears. Aortography confirmed the absence of intimal disruption. Medical therapy with beta and alpha blocking agents was established. After 5 months the patient was asymptomatic. On transesophageal echocardiogram and magnetic resonance, complete reabsorption of the hematoma of descending aorta was evident while a clearly identifiable false lumen and intimal flap (without tear) were present at the aortic arch level. The diagnostic criteria and therapeutic options are discussed and the literature on this topic is reviewed. PMID- 8402744 TI - [The variability of the heart rate in patients with a myocardial infarct undergoing systemic fibrinolysis interventions]. AB - Although heart rate variability is a very important prognostic factor in patients with myocardial infarction, the mechanism for the reduction in the vagal cardiac activity is unknown: myocardial infarction can cause local areas of sympathetic or parasympathetic denervation that leads to catecholamine hypersensitivity; the destruction of local ventricular receptors of the autonomic system may alter feedback to the higher centres, impairing autonomic regulation; the mechanical distortion consequent to the infarction can give rise to sympathetic overactivity. Effective thrombolysis in myocardial infarction could reduce myocardial necrosis and, therefore, cause less reduction of vagal activity and/or less sympathetic stimulation. To determine whether systemic thrombolysis has some effect on heart rate variability we studied 40 patients with a first transmural myocardial infarction. Enrollment criteria were very strict: all the patients with conditions potentially influencing heart rate variability were excluded from this study. Between day 15 and day 25 from the infarction patients had an Holter monitoring. A program we developed provided for the automatic identification of R wave, for the calculation of RR interval, and for the recognition of pause, extrasystolic and post-extrasystolic beats, that could be thus excluded from the analysis. Patients were divided in 2 groups: 17 patients treated with thrombolysis and 23 subjects treated traditionally. There were no difference between the 2 groups with regard to the age and cardiac function, except for the less echo score in the fibrinolytic treated group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402745 TI - Blood pressure, left ventricular hypertrophy and diabetes among 179 very old hypertensives from an Italian general population. The CASTEL (Cardiovascular Study in the Elderly). AB - Hypertension is common in elderly subjects, but old and particularly very old people have usually been excluded from major epidemiological trials. We studied 179 hypertensive subjects aged 80 years or more drawn from elderly people of an Italian town within the context of the CASTEL (Cardiovascular Study in the Elderly). Prevalence of hypertension declined from 66.7% (first visit, first measurement) to 56.3% (last visit, last measurement). Systolic but not diastolic blood pressure was a little higher among very old hyperglycemic hypertensive subjects than in normoglycemic ones, while left ventricular mass was independent of both blood pressure and glucose intolerance. PMID- 8402746 TI - [The effect of nifedipine in hypertensive cardiopathy. An echocardiographic and electrocardiographic study]. AB - To assess whether antihypertensive therapy by nifedipine can reverse left ventricular (LV) hypertrophy, 15 hypertensive patients, mean age 47 years, were serially studied during 12 months of treatment with nifedipine in slow release (40-60 mg/day), by recordings of blood pressure (BP), ECG and echocardiogram. Blood pressure decreased from 161 +/- 6/104 +/- 3 mmHg to 131 +/- 3/89 +/- 1 mmHg, p < 0.001, and this fall first became statistically significant at 1 month. From the hemodynamic view point, BP decreased for a reduction in total peripheral resistance. The Sokolow-Lyon voltage decreased significantly after 6 months (from 33.5 +/- 2.7 to 28.1 +/- 2.1 mm, p < 0.01) without further changes in the subsequent months. Left ventricular mass, by echocardiography, decreased after 6 months (from 189 +/- 15 to 176 +/- 13 g/m2, p < 0.05) and further after 12 months (169 +/- 13 g/m2, p < 0.001). The reduction in LV mass was secondary to the decrease in wall thickness, particularly in posterior wall thickness. No significant changes were observed in LV fractional shortening throughout the study. Thus, nifedipine was an effective antihypertensive agent and reverted LV hypertrophy secondary to arterial hypertension without impairment of LV systolic function. PMID- 8402747 TI - [The regression of left ventricular hypertrophy and improvement in diastolic filling during antihypertensive therapy with cilazapril]. AB - In order to evaluate the effects of cilazapril on left structure and function in essential hypertension, we evaluated 10 patients (4 females and 6 males) affected with mild-to-moderate systemic hypertension. All patients were treated with cilazapril 5 mg/day over a 6-months period and underwent a 24 hours ambulatory blood pressure monitoring and a complete Doppler echocardiographic examination at study entry, after 3 and 6-months of therapy. After therapy, mean systolic and diastolic arterial pressure decreased significantly from 153 +/- 16/102 +/- 8 mmHg to 135 +/- 13/83 +/- 6 mmHg respectively (p < 0.005/0.001 respectively). Moreover there was a significant decrease of left ventricular mass from 109 +/- 27 to 87 +/- 23 g/m2 (p < 0.005). Ejection phase indices did not change significantly, whereas left ventricular diastolic filling improved after therapy with a significant increase of M1/M2 ratio from 0.9 +/- 0.2 to 1.1 +/- 0.3 (p < 0.02). In conclusion, these data demonstrate that cilazapril 5 mg/day is effective in reducing blood pressure in mild-to-moderate essential hypertension. This normalization of blood pressure measurements is paralleled by a significant decrease of left ventricular mass index and by an improvement of left ventricular diastolic filling. PMID- 8402748 TI - [A validation of the data obtained with the simultaneous recording of blood pressure and the 24-hour electrocardiogram]. AB - Aim of this study was to evaluate the blood pressure (BP) measurement reliability of a light weight ambulatory BP and ECG recorder. Micro AM is a new 300 g portable apparatus that combines in one device both the ambulatory BP and solid state ECG recording. The dimensions of the Micro AM are 75 x 140 x 29 mm. The monitor measures BP using Korotkoff phase 1 for systolic and phase 5 for diastolic BP, and concurrently measures oscillometric BP, one method validating the other. In addition, the manual and programmed BP measurement modes can be supplemented by an "intelligent" mode in which the ECG triggers an ambulatory BP reading during an abnormal ST segment change. A standard mercury manometer was connected with the cuff of the Micro AM with a Y-shaped part, and 12 BP measurements were simultaneously taken at 5 min intervals by the automatic device in auscultatory mode and by a trained technician in 86 normotensive volunteers (aged from 18 to 44 years, 37 males and 49 females). The algebraic differences, the frequency distribution and the difference distribution of systolic and diastolic data between the 2 methods were calculated. The results show that the automatic method gives values for systolic BP that are lower than conventional ones (average differences -0.643 mmHg), whereas for diastolic BP, the values are higher (average differences +0.229 mmHg). Then, Student's paired t-test was used to evaluate statistically significant differences. The test relative to systolic BP was significant to the critical level of 0.1%, but the differences being 3 times smaller than the instrumental tolerance. On the contrary, diastolic BP differences were non significant. In conclusion, we found a good agreement between BP recorded automatically and by sphygmomanometer. PMID- 8402749 TI - Effects of a preload increase on ventricular filling in coronary artery disease. AB - This study was conducted to investigate the effects of a preload increase on ventricular filling dynamics in 50 coronary artery disease patients both before and immediately after leg elevation. Mitral and tricuspid peak E and A velocities and the mitral E/A velocity ratio increased after leg elevation whereas isovolumic relaxation time and mitral deceleration time decreased. The increase in tricuspid peak A velocity after leg elevation was related to the positivity of the exercise stress test (p = 0.01). The increase in mitral peak A velocity was related to the history of old myocardial infarction (p = 0.006). PMID- 8402750 TI - Effect of temperature and coronary flow on the metabolic and mechanical function of the isolated rat heart. AB - A number of cardiac metabolic intermediates, namely, adenosine triphosphate (ATP), H+, phosphocreatine (PCr), inorganic phosphate (Pi), adenosine diphosphate (ADP), and related functions of these intermediates, Gibbs' free energy of ATP hydrolysis (delta G) and phosphorylation ratio [ATP/(ADP.Pi)], are thought to adjust mitochondrial oxidative phosphorylation rates to conform to mechanical demand. The effects of hypothermia and altered perfusion pressure on these parameters were evaluated in 12 hearts from Sprague-Dawley rats perfused in the Langendorff mode. 31P-nuclear magnetic resonance (NMR) spectra were obtained at cardiac temperatures between 20 and 37 degrees C, and coronary perfusion pressures between 20 and 145 cm H2O. Coronary flow varied between 0.5 and 15 ml/min throughout this range of intervention. Heart rate (HR), left ventricular systolic pressure (LVSP), and specific volumetric coronary flow (SCF) were determined for each temperature and perfusion pressure. The product HR x LVSP directly correlated with perfusion pressure at all temperatures. The temperature dependence could be represented by an overall activation energy of 72.7 kJ/M. In the constant temperature experiment, SCF and HR x LVSP fell linearly with decreasing perfusion pressure. Quantitative evaluation of the relationship between cardiac function and the metabolic intermediates described above defined these intermediates as nonregulatory with the possible exception of H+. PMID- 8402751 TI - Hemodynamic and biochemical changes after chronic administration of cilazapril to hypertensive patients. AB - The study describes the changes in basic hemodynamic parameters after long-term antihypertensive therapy with cilazapril--a new ACE inhibitor lacking a sulfhydryl group--in hypertensive patients and the drug effects on renal function, glucose tolerance and lipid metabolism. 30 patients (18 males, 12 females, mean age: 53.3 +/- 18 years) with mild to moderate essential hypertension were studied. The following determinations were performed in patients, before and after 4.5 months of cilazapril monotherapy at a dose of 5 mg/24 h: (a) antihypertensive action of the drug (arterial pressure at rest and during a 24-hour recording); drug effects on left ventricular (LV) mass index; its contractility indexes (%FS, EF) and the left atrial emptying index were studied by means of echocardiography; (b) plasma insulin concentration during oral glucose tolerance tests, in the fasting state, after the administration of 75 g glucose per os, as well as the changes in the insulinogenic index and the 6 keto-PGF1 alpha/TXB2 ratio, and (c) drug effect on renal function (urea, creatinine, uric acid, plasma electrolytes), blood lipid profile (total cholesterol, triglycerides, HDL-CH) and serum transaminases. Long-term drug administration exhibits an effective antihypertensive action, without causing reflex tachycardia and also reduces the LV mass index without affecting its EF, while improving its diastolic function. It does not significantly affect the various biochemical parameters, and achieves glucose regulation, both in the fasting state and after glucose loading, with a decrease in the insulinogenic index, and simultaneously increases the 6-keto-PGF1 alpha/TXB2 ratio. The existence of a direct cause-effect relationship between the changes in the above hormone systems is possible. PMID- 8402752 TI - Increase in left ventricular ejection rate during recovery from exercise in patients with myocardial infarction. AB - Changes in left ventricular (LV) ejection rate were evaluated during supine bicycle exercise and recovery in 12 patients with anterior myocardial infarction. Pulmonary artery wedge pressure (34 +/- 9 to 14 +/- 5 mm Hg) and plasma norepinephrine level decreased at 2 min recovery from peak exercise, whereas plasma lactate tended to increase. As a result, LV ejection rate reached the highest value at 2 min of recovery. Thus, both cardiac (optimal filling pressure) and peripheral factors (reduced vascular resistance) caused the increase in LV ejection rate at early recovery. PMID- 8402753 TI - Reocclusion following successful thrombolysis. Emerging concepts. AB - Thrombolytic therapy, through the restoration of coronary arterial blood flow and myocardial perfusion significantly improves outcome among patients with acute myocardial infarction. Despite its widely appreciated benefits, however, thrombolysis removes only a small portion of existing thrombus at the site of atheromatous plaque rupture. Further, thrombogenic substrate is frequently exposed and may even be generated to the extent that rethrombosis occurs, causing recurrent ischemia, reinfarction, and coronary reocclusion. The mechanisms underlying this important event are discussed. PMID- 8402754 TI - Prolonged activation of fibrinolytic system induced by fibrin nonselective thrombolytic agent can contribute to preventing early reocclusion after coronary thrombolytic therapy. AB - The incidence of early reocclusion is reported to be higher in patients who receive fibrin-specific thrombolytic agents than nonspecific ones. The reason has yet to be clarified. In the present study, we focused on the difference in duration of fibrinolytic activity. The hemostatic parameters of 7 consecutive patients suffering from acute myocardial infarction treated with a fibrin nonspecific thrombolytic agent (urokinase) were compared with 9 patients who received a fibrin-specific agent (tissue plasminogen activator, t-PA). The plasma concentrations of alpha 2-plasmin inhibitor (alpha 2-PI), plasmin alpha 2-PI complex (PLC), fibrin degradation products E fragment (FDP-E), and D-D dimer (D dimer) were measured before, soon after, 1, 2, 3, 4, and 6 h and 2, 3, 4, and 7 days after thrombolytic therapy to estimate the hemostatic and fibrinolytic state. A significant decrease in alpha 2-PI (less than the lowest measurable level) with a simultaneous increase in FDP-E and D-dimer was induced soon after the administration of urokinase. FDP-E and D-dimer decreased, with a significant increase in alpha 2-PI, more than 6 h after thrombolytic therapy. In contrast, a less significant decrease in alpha 2-PI with a lesser amount and shorter duration of fibrinolysis were observed in patients who received t-PA. The amount of PIC soon after drug administration was not different between the two groups. Our data suggested that fibrinolytic activities induced by fibrin-nonspecific urokinase persisted longer than expected by its plasma half-life.The fibrinolytic activities might be terminated by the production of alpha 2-PI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402755 TI - Serial changes in plasma concentration of plasminogen activator inhibitor-1 before and serially after thrombolytic therapy for acute myocardial infarction. AB - Plasminogen activator inhibitor-1 (PAI-1), which is secreted from vascular endothelial cells, plays an important role in regulating fibrinolysis. We measured the plasma concentrations of tissue-type plasminogen activator (t-PA), PAI-1 and t-PA PAI complex before and serially after thrombolytic therapy for acute myocardial infarction to clarify the relationship between thrombolytic therapy and PAI-1. Plasma concentrations of t-PA, PAI-1 and t-PA PAI complex before thrombolytic therapy were 11.6 +/- 5.5, 27.0 +/- 13.0 and 7.3 +/- 5.4 ng/ml, respectively. t-PA and t-PA PAI complex increased to 25.0 +/- 5.3 and 14.3 +/- 6.5 ng/ml immediately after drug administration; however, the level of PAI-1 decreased slightly immediately after thrombolytic therapy. The PAI-1 level increased again several hours after therapy, especially in patients showing apparently successful reperfusion. All values returned to normal 4-7 days after thrombolytic therapy. Not only augmented antifibrinolytic activity suggested by increased PAI-1 but also augmented fibrinolytic activity suggested by increased t PA was observed in patients with acute myocardial infarction. These abnormal findings persisted several days after coronary thrombolytic therapy. PMID- 8402756 TI - Cardiac amyloidosis presenting as ischemic heart disease. A case report and review of literature. AB - Cardiac amyloidosis can have varied clinical presentations, but it uncommonly leads to typical angina. The exact cause of ischaemia in this condition is not known, though various mechanisms have been postulated. We describe here a case of cardiac amyloidosis, proven by endomyocardial biopsy, which presented as ischaemic heart disease, and we also review the literature on the same. PMID- 8402757 TI - Noninvasive diagnosis of reperfusion in acute myocardial infarction. PMID- 8402758 TI - Early electrocardiographic changes in acute myocardial infarction treated by streptokinase or alteplase: a randomized study with dynamic, multi-lead, electrocardiographic monitoring. AB - Patients with a patent coronary artery after reperfusion therapy show an increased rate of evolution of QRS changes and of resolution of S-T changes as compared to patients with a persistent occlusion. Dynamic vectorcardiography permits multi-lead monitoring of changes in QRS complex and S-T segment over time. We monitored 150 patients randomized to alteplase or streptokinase for 24 h after admission. Alteplase was associated with significantly more rapid evolution of electrocardiographic changes but also with an increased occurrence of recurrent S-T changes. This may indicate a higher rate of early reperfusion with alteplase but due to recurrent ischemia, an electrocardiographic catch-up is seen with similar extent of myocardial damage observed after 24 h of monitoring. PMID- 8402759 TI - Evidence for plasma-mediated neutrophil superoxide anion production during myocardial infarction. AB - The participation of polymorphonuclear neutrophils (PMN) in the development of free-oxygen-mediated myocardial injury is well documented, but direct evidence that PMN-oriented stimuli released to the peripheral blood are able to stimulate PMN free oxygen radical production is missing. We have previously reported that peripheral blood plasma obtained from patients with acute myocardial infarction has chemotactic activity for neutrophils and augments PMN adherence. To investigate whether neutrophilic stimuli released to peripheral blood may induce PMN superoxide anion (O2-) production, we incubated PMN from healthy donors with plasma from patients with acute transmural infarction. PMN O2- production was measured by cytochrome c reduction. PMN O2- was higher under the influence of plasma obtained on the day of admission (24.66 +/- 12.41) and 1 day after onset of acute ischemia (22.91 +/- 10.37) as compared with those observed after incubation with saline (4.18 +/- 1.37; negative control) or zymosan-activated plasma (11.07 +/- 3.4; activated complement cascade positive control). In the following days, plasma-mediated PMN O2- decreased: 13.27 +/- 1.96; 12.37 +/- 3.54 and 8.18 +/- 1.56 after incubation with plasma obtained 2, 3 and 7 days after onset of symptoms, respectively. The results indicate an additional possibility of monitoring the inflammatory response to myocardial necrosis. PMID- 8402760 TI - Venoarterial carbon dioxide tension gradient in acute heart failure. AB - The venoarterial carbon dioxide tension gradient (P[v-a]CO2) was studied in patients with acute myocardial infarction. Seven patients with congestive heart failure (CHF group) and 10 patients without heart failure (control) were enrolled in this study. In all patients, hemodynamics were continuously monitored. Simultaneously, arterial and mixed venous blood were sampled, and blood gases and lactate concentration were analyzed. At the initial measurement before therapy, arterial and mixed venous pH and bicarbonate values were within the normal range, and there was no significant difference between the CHF group and controls. There was also no difference in arterial oxygen tension under the differential conditions of oxygen inhalation. However, cardiac index and mixed venous oxygen saturation (SvO2) were significantly lower, while the oxygen extraction ratio (OER) and arterial lactate were significantly higher in the CHF group than in the controls. On the other hand, P[v-a]CO2 was significantly higher in the CHF group (7.8 +/- 2.6 vs. 3.5 +/- 2.2 mm Hg, p < 0.01). This finding was due to the elevated mixed venous carbon dioxide tension in the CHF group, since arterial carbon dioxide tension was the same in both groups. Analysis of a total of 42 measurements obtained during the therapeutic course in the CHF group revealed a correlation of P[v-a]CO2 with cardiac index (r = -0.3, p < 0.05), OER (r = 0.57, p < 0.001), SvO2 (r = -0.56, p < 0.001) and lactate (r = 0.62, p < 0.001). The increase in P[v-a]CO2 observed in acute heart failure suggests the evidence of intracellular acidosis despite the absence of acidemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402761 TI - Protective effect of triflusal against acute myocardial infarction in patients with unstable angina: results of a Spanish multicenter trial. Grupo de Estudio del Triflusal en la Angina Inestable. AB - A multicenter, double-blind, placebo-controlled study was carried out to evaluate the effect of a new antiplatelet agent, triflusal (2-acetoxy-4-trifluoromethyl benzoic acid), in the prevention of nonfatal myocardial infarction and cardiac or vascular death (principal end-points) in patients with unstable angina. 281 patients were randomly assigned to triflusal (300 mg t.i.d.; n = 143) or placebo (n = 138). After 6 months of treatment, the incidence of nonfatal acute myocardial infarction was significantly lower in the triflusal than in the placebo group: 6 patients (4.2%) versus 17 (12.3%), p = 0.013. The low number of deaths (2/143 triflusal versus 0/138 placebo recipients) hampered statistical analysis of mortality rates. The need for revascularization was similar in the two groups: 24 patients (16.8%) in the triflusal group and 28 (20.3%) in the placebo group, p = 0.449. In conclusion, the results show that treatment with triflusal can reduce the incidence of myocardial infarction in patients with unstable angina. PMID- 8402762 TI - The effect of short-term celiprolol therapy on platelet function in essential hypertension. AB - The reduction of increased platelet aggregation in essential hypertension is one of the aims of modern antihypertensive therapy. Twenty-one hospitalized patients with non-treated essential hypertension were examined. The platelet function measurements were made before the therapy and after 1 week of celiprolol administration (300 mg/day). Fifteen essentially hypertensive patients were investigated before and after 1 week of placebo administration. Plasma beta thromboglobulin was assayed, and the whole blood platelet aggregation (initial and total-induced by adrenaline and ADP) was measured. A significant decrease in adrenaline-induced (from 19 to 13%, p < 0.02) and ADP-induced aggregation (from 15 to 13%, p < 0.05) was observed after celiprolol administration. This reduction of adrenaline-induced platelet aggregation may be explained by the stimulation effect of celiprolol on platelet beta 2-receptors. Thus, some inhibitory effect of celiprolol on platelet aggregation is one of the further advantages of this drug in the therapy of essential hypertension. PMID- 8402763 TI - Effect of prolonged nifedipine or captopril therapy on lymphocyte magnesium and potassium levels in hypertension. AB - The effect of prolonged treatment with calcium channel blockers on potassium and magnesium stores is uncertain. We measured lymphocyte and serum magnesium and potassium in 28 patients treated for hypertension for 6 months with nifedipine or captopril. There was no difference in serum or lymphocyte concentrations in the two groups compared to 45 healthy, normotensive controls. These results suggest that intracellular cation levels are maintained with prolonged therapy with calcium channel blockers. PMID- 8402764 TI - Structural basis for mitral valve dysfunction associated with ostium secundum atrial septal defects. AB - Mitral valve (MV) specimens obtained at operation from 50 patients with ostium secundum atrial septal defect and associated MV dysfunction were studied macro- and microscopically to evaluate the structural lesions. The functional abnormalities in these patients were isolated mitral stenosis, isolated mitral regurgitation, mitral valve prolapse or mitral stenosis with regurgitation. Post inflammatory lesions were commonly found in those with mitral stenosis who had a past history of rheumatic fever. Patients with mitral valve prolapse had either post-inflammatory lesions or lesions attributable to haemodynamic stress but different from the extensive lesions seen in floppy MV syndrome. PMID- 8402765 TI - Myocarditis masquerading as ischemic heart disease: the diagnostic utility of antimyosin imaging. AB - The diagnosis of myocarditis presents a diagnostic challenge due to its varied clinical presentation. In addition, criteria for myocarditis are varied. At present, the confirmation of myocarditis depends on an endomyocardial biopsy demonstrating myocardial inflammation and necrosis. Unfortunately, this invasive procedure is associated with some degree of risk and has significant limitations. This report discusses the case presentations of two patients with chest pain, electrocardiographic changes and elevated creatine kinase levels suggestive of myocardial infarction, who were subsequently found to have findings compatible with myocarditis based on indium-111 antimyosin antibody scanning. This noninvasive test therefore appears to have value in the differentiation of myocardial ischemia from myocarditis. PMID- 8402766 TI - Transesophageal echocardiography. Indications and technical considerations. AB - The superior imaging capabilities of TEE have rapidly thrust this technique into the mainstream of noninvasive cardiology. However, the semi-invasive nature of this procedure requires specialized training on the part of the echocardiographer and adaptations of the traditional echocardiographic laboratory. These requirements will become even more evident as this technique is employed increasingly for interventional studies such as transesophageal atrial pacing and pharmacologic stress. TEE has proved efficacious and safe, even in critically ill patients, and its applications continue to expand. Following an article on anatomy, encompassing single and biplane orientation, the remainder of this monograph addresses the established as well as the emerging applications of TEE. PMID- 8402767 TI - Single, biplane, multiplane, and three-dimensional transesophageal echocardiography. Echocardiographic-anatomic correlations. AB - Advances in TEE instrumentation have led to the emergence of this modality as a powerful diagnostic tool. Numerous investigations in humans have clearly defined the plethora of clinical applications of TEE in a variety of patient care scenarios. Research in progress in the area of transducer technology and computerized image processing is likely to bring 3-D TEE to the clinical front. PMID- 8402768 TI - Intraoperative assessment of left ventricular function with transesophageal echocardiography. AB - Qualitative TEE assessment is used to guide administration of fluids and inotropic drugs and to monitor left ventricular function intraoperatively. Left ventricular hypovolemia or depression is easily recognized by directly noting a small end-diastolic area or low ejection fraction. Appropriate therapy can be instituted and continuously monitored. In contrast, pulmonary artery pressure monitoring does not accurately indicate loading conditions during major cardiovascular procedures or whenever left ventricular compliance is impaired, mitral valve dysfunction is present, or right ventricular distention occurs. New applications and technical improvements in TEE are being developed at a remarkable rate. Future versions of ABD technology are likely to address the problem of anisotropy, require less user intervention, and incorporate 3-D information from multiplane probes to produce real-time estimates of left ventricular volumes. The raw information in the returning signal will most likely be further analyzed to allow characterization of ischemic but still viable tissue. Coupled with the ability to assess regional myocardial perfusion by contrast echocardiography, the clinician will be able to institute more timely and appropriate medical and surgical therapy. TEE assessment of mitral valve function has become the standard of care after mitral valve repair, and in a similar fashion, assessment of myocardial perfusion by TEE may become the standard of care during cardiac and major noncardiac surgery. PMID- 8402769 TI - Transesophageal echocardiography in the evaluation of mitral regurgitation. The twenty-four signs of severe mitral regurgitation. AB - Many echocardiographic signs of severe MR are clearly demonstrated, particularly when both TEE and TTE are used. When these signs are assiduously sought, the recognition of severe MR should pose little problem. Part of the confusion concerning MR and the grading of its severity comes from the fact that the hemodynamic consequences of a given degree of MR vary widely from one individual to another. A regurgitant volume of 50 mL might prove incapacitating to one patient while seeming inconsequential in a second patient. A regurgitant fraction of 50% is poorly tolerated in some patients and asymptomatic in others. Similarly, a regurgitant orifice 0.5 cm2 has unpredictable consequences to the organism, and, in fact, this orifice may vary considerably in size depending on hemodynamic conditions. Thus, a universal definition of the severity of MR is lacking, and there is no agreement on the units with which to quantitate it. The net effect of this confusion is not an inability to recognize severe MR but frustration in differentiating moderate MR from severe MR. We believe that precise quantitation of MR will occur when comprehensive pharmacologic interventions with either TEE or surface echocardiographic monitoring are performed to define the severity of MR by its range of responses to these agents. We have had some success with Doppler measurement of the response of pulmonary artery pressure to dynamic exercise. Patients with normal pulmonary artery pressure at rest tend to show exaggerated rises in pulmonary pressure when MR is clinically important and has resulted in left ventricular dysfunction. Anticipated progress notwithstanding, competently performed TEE is the method of choice for recognizing severe MR. PMID- 8402770 TI - Mitral stenosis. Evaluation and guidance of valvuloplasty by transesophageal echocardiography. AB - Percutaneous transvenous balloon mitral valvuloplasty is a well-established alternative to surgical commissurotomy for selected symptomatic patients with rheumatic mitral stenosis. This procedure normally is performed with fluoroscopic guidance; however, even the experienced operator can be misled by radiographic landmarks. The addition of on-line TEE during PBMV facilitates the success and safety of this procedure by guiding the transseptal puncture and the positioning of the dilating balloon catheters across the stenotic mitral valve. In addition to guiding the manipulation of catheters, on-line TEE is also useful for confirming the efficacy of valvuloplasty and for detecting complications. TEE during PBMV is feasible, safe, and well-tolerated by awake patients. PMID- 8402771 TI - Transesophageal echocardiography in the evaluation of prosthetic valves. AB - Biplane TEE, with its relatively low risk, has become an invaluable adjunct to comprehensive transthoracic 2-D Doppler and color flow examination. It should be undertaken in all instances in which transthoracic information is not adequate, or if there is suspicion of an abnormality not detected by TEE. One can advocate the use of TEE in the operating room in all instances wherein a patient is scheduled to undergo implantation of a prosthetic valve, because it enables baseline information to be obtained that may be valuable in the management of patients with questionable findings, permitting a comparison. The role of cardiac catheterization is limited. Hemodynamic cardiac catheterization should only be resorted to in instances in which there is discrepancy between echocardiographic findings and clinical presentation. Visualization of coronary arteries may be indicated in some situations in which a limited coronary arteriogram could be obtained. This procedure is less morbid if separated from hemodynamic catheterization of the right and left sides of the heart. Echocardiography represents the state-of-the-art in the evaluation of patients with valvular prostheses. Biplane TEE and multiplane TEE are valuable complementary technologies to TEE, when the situation dictates its use, because these modalities provide useful information that alters management of the patient. PMID- 8402772 TI - Transesophageal echocardiography in endocarditis. AB - TEE plays a central role in the diagnosis and management of infective endocarditis. In experienced hands this technique is probably over 90% sensitive and specific for the detection of intracardiac lesions associated with endocarditis. TEE should be performed as soon as possible once endocarditis is suspected. A technically adequate negative study almost always means a low probability of endocarditis. Transesophageal imaging appears to be an accurate method of detecting leaflet perforation as well as abscess and fistula formation. PMID- 8402773 TI - Systemic arterial emboli and cardiac masses. Assessment with transesophageal echocardiography. AB - Several cardiac and aortic findings on TEE have been implicated as sources for systemic emboli. These findings include left atrial thrombus, left atrial spontaneous contrast, patent foramen ovale, atrial septal aneurysm, and aortic atheroma. The prevalence of each of these abnormalities has been significantly greater in patients with suspected embolic events as compared with controls. Furthermore, the sensitivity of TEE for these findings is much greater than that of TTE. Thus, patients with unexplained cerebral ischemia or systemic emboli should undergo TEE so that possible sources can be detected. In regard to cardiac tumors, although the data are preliminary, certain tumor locations and morphologic aspects are better evaluated with TEE than with other techniques. More importantly, it has been suggested that a change in management of cardiac tumors is initiated by TEE in the majority of instances. TEE provides important diagnostic information in many patients with suspected cardiac emboli and cardiac masses and should be strongly considered in the work-up of such patients. PMID- 8402774 TI - Role of transesophageal echocardiography in dissection of the aorta and evaluation of degenerative aortic disease. AB - The combination of different ultrasound techniques such as transthoracic, suprasternal, subcostal, and TEE has a high sensitivity and specificity in the diagnosis of aortic dissection. Limitations of this combined ultrasound technique are related to the visualization of the ascending part of the aortic arch, which, because of the interposition of the trachea, cannot be visualized completely. The beginning or end of a dissection in this part of the aorta may be misinterpreted. However, false-negative results are rare. False-positive results due to artifacts resulting from reverberations in an ectatic ascending aorta must be taken into account. The most important diagnostic goals in acute or chronic aortic dissection are (1) confirmation of the diagnosis by visualization of the intimal membrane; (2) the differentiation of true and false lumen, depending on visualization of spontaneous echocardiographic contrast, thrombus formation, slow or reduced reversed flow, systolic diameter reduction, and signs of entry jet into the false lumen; (3) detection of intimal tear, demonstrating communication by 2-D or color Doppler echocardiography; (4) determination of the extent of dissection with classification according to DeBakey types I, II, and III, or Stanford types A and B, with differentiation between communicating or noncommunicating dissection and antegrade or retrograde dissection limited to the descending aorta or expanding into the ascending aorta; (5) detection of wall motion abnormalities as a sign of preexisting coronary artery disease or myocardial ischemia due to ostium occlusion by an intimal flap, coronary artery rupture, or collapse of the true lumen during diastole; (6) detection and grading of aortic insufficiency; (7) detection of side branch involvement by suprasternal, subcostal, and abdominal sonography (which will provide information about the choice of the site for cannulation or catheterization of the femoral artery); and (8) detection of pericardial or pleural effusion and mediastinal hematoma as signs of an emergency situation (i.e., suspending rupture). Based on ultrasound diagnostic information, operation can be performed in all acute situations in patients with type A dissection without further investigation. The ability to act decisively in this setting is particularly important in patients with signs suggesting a dire prognosis (i.e., pericardial or pleural effusion or mediastinal hematoma). For follow-up studies, the combination of echocardiography with MR tomography is recommended. With TEE, entry tears can be detected with a higher sensitivity than with MR tomography. This capability may be important for the patient's prognosis. MR tomography, on the other hand, has a better spatial resolution showing the entire aorta, particularly the ascending aortic arch. PMID- 8402775 TI - Hemodynamics by transesophageal echocardiography. AB - The transesophageal approach has improved echocardiographic investigation of cardiac structure and function. As a new window to the heart with markedly improved resolution, TEE gives better insight into cardiac morphology and pathology than does precordial imaging. Specifically, the LA and mitral valve can be better visualized due to the immediate retrocardiac position of the imaging transducer. Similarly, TEE is also widely used to estimate left ventricular functional status. Specifically in the perioperative setting, methods have been developed and tested to analyze global and regional left ventricular function. In addition, methods of estimating left atrial pressure (pulmonary capillary wedge pressure) have recently been developed using pulsed Doppler echocardiography of pulmonary venous flow and interatrial septal dynamics. Transesophageal pulsed Doppler echocardiography of pulmonary venous flow provides a useful clinical tool to estimate pulmonary capillary wedge pressure reliably in the setting of impaired systolic function, diastolic dysfunction, or both. Furthermore, pulmonary venous flow is characteristically altered in patients with severe MR and can be useful in grading its severity. Transesophageal continuous wave Doppler echocardiography may prove useful to estimate systolic pulmonary artery pressure as another clinically useful hemodynamic parameter. Therefore, TEE adds significantly to the noninvasive assessment of cardiac hemodynamics. PMID- 8402776 TI - Transesophageal echocardiography in the critical care patient. AB - The superior resolution of TEE has substantially improved the diagnostic capabilities of the echocardiographer. In the critical care setting in which TTE is frequently extremely limited, TEE is proving to be an invaluable tool. The immediate availability of morphologic and hemodynamic information permits the cardiologist and intensivist to institute rapid changes in clinical management. Further miniaturization of transducers and probes as well as the ongoing development of automated techniques may eventually lead to the use of TEE as a monitoring device in the critical care patient. To date, there is no proof that application of this technique has improved overall survival in this group of patients. However, anecdotal experience and recently published series suggest that otherwise inapparent diagnoses have been made, leading to overall clinical improvement. PMID- 8402777 TI - Transesophageal echocardiography in congenital heart disease in the adult. AB - Progress achieved during the last three decades in medical and surgical treatment of infants and children with congenital heart disease has resulted in a significant number of patients surviving to adulthood. The complexity of the basic malformations in addition to the superimposed surgical interventions makes accurate diagnostic imaging essential. The scope of information obtainable from high-quality TEE supports its use in the follow-up of operated and unoperated adult patients, as well as in surgical and interventional procedures. Application of the segmental analysis of anatomy is as important during TEE as it is during TTE. Supplemental transthoracic 2-D and spectral Doppler flow measurements may be needed when poor beam alignment during TEE precludes accurate gradient determinations. Although, to date, most reports of TEE in patients with congenital heart disease have involved children and adolescents, the informational advantage is potentially greater in older adolescents and adults in whom transthoracic windows are frequently limited. In our experience with 125 TEE studies in adult patients with congenital heart disease (the majority using a biplane probe), horizontal plane TEE imaging provided important additional information concerning pulmonary venous connections, atrial baffle function, atrioventricular anatomy and function, left ventricular outflow tract lesions, and great artery positional relationships. Vertical plane TEE enhances imaging of systemic and pulmonary venous connections, atrial situs, venosus atrial septal defects, atrioventricular anatomy and function, and the ventricular septum and outflow tracts and is essential for complete assessment of complex right ventricular outflow tract anatomy, ventriculoarterial alignment, and the sizing of all aortic segments in coarctation of the aorta. PMID- 8402778 TI - Use of transesophageal echocardiography in evaluating coronary arteries. AB - Because of the better resolution of higher frequency transducers and the proximity of the coronary arteries to the esophageal window, TEE is emerging as a valuable method for evaluation of coronary artery disease. TEE allows imaging of the proximal coronary arteries, measurement of coronary flow reserve, identification of coronary artery anomalies, and observation of wall motion during transesophageal atrial pacing. The application of TEE in evaluation of coronary artery disease will continue to grow as technology improves. PMID- 8402779 TI - Transesophageal echocardiography during cardiopulmonary resuscitation. AB - Transesophageal echocardiography is ideally suited for imaging during CPR because high-quality images can be obtained immediately and continuously without interruption of cardiac compression and ventilation. Use of TEE during CPR is increasing to help monitor resuscitative efforts, for diagnosis, to assist in understanding the physiology of blood flow, and for evaluation of new methods of CPR. PMID- 8402780 TI - Infrarenal aortic stents: initial clinical experience and angiographic follow-up. AB - From March 1990 to May 1991, arterial stents were placed in seven patients because of a tight stenosis (five patients) or a total chronic occlusion (two patients) located in the infrarenal aorta. In one case, the aortic occlusion extended to both common iliac arteries. After balloon dilatation, aortic stents were successfully positioned in all cases. Bilateral common iliac recanalization and stent placement were performed in one case. No complications occurred in any of the patients. No complications occurred in any of the patients. Follow-up data were derived from clinical assessments and angiographic results. After a 15.1 month mean follow-up period (range 12-24 months), the seven aortic stents remained patent. Three iliac artery procedures were performed in two of the patients as well. Claudication recurred in three of the seven patients which was related to a common iliac occlusion (one case) or distal progression of atherosclerosis (two cases). Aortic stents seem to be suitable for treating failed angioplasty of aortic lesions but the procedure remains technically difficult when there is associated severe atherosclerosis of the proximal common iliac arteries. Nevertheless, considering the morbidity rate (0%) and the patency rate in this series, this technique could become an alternative to surgical treatment for infrarenal aortic occlusive lesions. PMID- 8402781 TI - Lipiodol retention and massive necrosis after lipiodol-chemoembolization of hepatocellular carcinoma: correlation between computed tomography and histopathology. AB - This retrospective study examined the computed tomography (CT) criteria for judging the effectiveness of transcatheter arterial Lipiodol-chemoembolization (Lp-chemo-TAE) in 35 cases with hepatocellular carcinoma (HCC). Massive necrosis, defined as involving 97% or more of the HCC nodule, was observed in 15 cases after Lp-chemo-TAE, whereas nonmassive necrosis, defined as involving < or = 96% of the HCC nodule, was observed in the remaining 20 cases. In 12 of 15 cases (80%) with massive necrosis, uniform dense retention of Lipiodol (Lp) was observed throughout the HCC nodule on CT images 3-4 weeks after Lp-chemo-TAE as opposed to only one (5%) of 20 cases with nonmassive necrosis (p < 0.01). Eight of nine cases (89%) with massive necrosis had tumor attenuation values of 365 Hounsfield units (HU) or greater on CT images 3-4 weeks after embolization, as opposed to only four (27%) of 15 cases with nonmassive necrosis (p < 0.01). We conclude that the effectiveness of the Lp-chemo-TAE can be judged on CT from the degree and duration of Lp retention in the HCC nodule and the measurement of the attenuation value of the HCC nodule. PMID- 8402782 TI - Percutaneous transluminal angioplasty of proximal subclavian artery occlusions. AB - Percutaneous transluminal angioplasty (PTA) is a well-established treatment for patients with subclavian artery stenosis and brachial or cerebral symptoms. Its efficacy for treatment of subclavian occlusions, however, is not yet established. We attempted to recanalize the subclavian artery in 46 patients with proximal subclavian artery occlusion and were successful in 38 (83%). In two patients, residual stenosis, and in five patients, thrombotic material on the arterial wall required subsequent implantation of a self-expandable stent. Two additional patients were left with residual stenoses because stents were not available at that time. There were no cerebral or brachial complications. During a mean follow up of 33 months, occlusion recurred in one patient after 3 months and stenosis recurred in five. PMID- 8402783 TI - Coronary artery fistulae without audible murmur in adults. AB - Retrospective analysis of 4886 adults undergoing coronary arteriography for evaluation of angina between October 1988 and December 1991, revealed coronary artery fistulae in eight patients (all men, aged 36-69 years). No murmur was audible in any of these eight patients. Associated significant coronary artery disease was detected in five patients. The feeder arteries to the fistula were both the left main coronary artery and the left anterior descending artery (LAD) in two, the LAD in six, and the right coronary artery in two patients. The fistula terminated in the pulmonary artery in seven patients and in the right atrium in one patient. Successful operative treatment (coronary artery bypass grafting and ligation of the fistula) was undertaken in four patients with severe obstructive coronary artery disease with satisfactory results. Follow-up for up to 2 years of the three patients with coronary artery fistula and no associated coronary artery disease who did not undergo surgery revealed continuing good prognosis. We conclude that coronary artery fistula in adults is a distinct, though rare (incidence in present series 0.11%) entity, without audible murmur, commonly associated with coronary artery obstructive disease, and that the diagnosis is mostly incidental during routine coronary arteriography. PMID- 8402784 TI - New retrievable percutaneous vena cava filter: experimental in vitro and in vivo evaluation. AB - A new retrievable percutaneous vena cava filter was tested in vitro and in vivo in 15 foxhounds. In vitro, the new vena cava filter was compared with the standard Kimray-Greenfield filter and the Gunther basket filter. The new filter is a stainless steel half-basket filter and is suitable for percutaneous antegrade or retrograde insertion through a 8.5 Fr introducer sheath. In vitro testing showed the filter causing no significant flow alterations and being highly effective in capturing medium- and large-sized thrombi; furthermore, fatigue testing revealed no breakage of the new filter, whereas the Gunther basket filter showed breakage of the struts. In vivo studies showed no occlusion, major perforation, or filter migration during follow-up of 2 weeks to 6 months. Tilting of the filter postimplantation occurred in two out of 28 filters. Ten of 11 filters were successfully retrieved by the transjugular approach 2 weeks after implantation. The device seems to be suitable for temporary or permanent protection against pulmonary embolism. PMID- 8402785 TI - Occlusive effect of metallic stents on canine ureters. AB - A three-part study, with successive modifications based on preceding results, was conducted to evaluate ureteral placement of metallic stents. Gianturco self expanding (10 mm and 4 mm diameter) and balloon-expanded (4 mm diameter) metallic stents were placed in normal and stenotic canine ureters. No migration or ureteral perforation occurred during the follow-up of 10 mm stents. Varying degrees of hydronephrosis and hydroureter were found on all 1-week pyelograms. At 4 weeks, complete occlusion of the stented ureter was noted in all cases because of mucosal hyperplasia around the stent wires. To prevent this reaction, 4 mm self-expanding stents constructed of smaller wire that was uncoated or coated with either Teflon or poly-urethane were tested in five dogs. In all cases, results were similar to those obtained with the larger prostheses. Finally, 4 mm balloon-expanded stents were placed in a normal ureter of three dogs. In one dog, the stent migrated out of the ureter. No migration or ureteral perforation occurred in the two remaining dogs. In these animals, mucosal hyperplasia and complete ureteral occlusion occurred 6 and 8 weeks after placement. Therefore, ureteral placement of Gianturco self-expanding as well as balloon-expanded metallic stents leads to occlusion of the ureter instead of maintaining its patency. Stents, therefore, may be useful as ureteral occlusion devices. PMID- 8402786 TI - Evaluation of a prototype steerable angioscopic laser catheter in a canine model: a feasibility study. AB - To overcome some of the persisting technical problems related to laser angioplasty, a new catheter was designed and investigated in a canine model. This 5F catheter contained a deflectable tip for steerability, an angioscope, and a laser fiber. Catheter steerability, angioscopic function, and the effects of a 480 nm flash lamp pumped pulsed dye laser on normal canine vessel walls were evaluated. Steering, angioscopic guidance, and application of laser energy were easy and fast to perform in a bloodless vessel segment. Maintaining a condition of bloodlessness at the target site, critical to angioscopic guidance, proved to be the most difficult part in this prototype evaluation. It was noted that the 480 nm pulsed dye laser did not cause macroscopic alterations or perforations to the normal vessel wall. We conclude that a relatively simple deflection mechanism of a small-caliber angioscope provides the kind of aiming ability requisite for precise endovascular therapy. Complete bloodlessness of the area is necessary for both viewing and laser ablation at 480 nm. PMID- 8402787 TI - Percutaneous fragmentation of pulmonary emboli in dogs with the impeller-basket catheter. AB - A new catheter system for rapid percutaneous fragmentation of pulmonary emboli was tested in nine dogs. The system consisted of a high-speed rotating impeller, placed within the center of a self-expandable basket. Preformed radiopaque emboli were introduced via the right external jugular vein. In nine tests in five dogs, a single embolus was launched after the right pulmonary artery had been balloon occluded and the impeller-basket catheter had been positioned into the left pulmonary artery. Seven of nine emboli were completely fragmented, each within less than 10 s. In two tests, performed in two different dogs, the catheter was positioned after complete embolization of the left pulmonary artery. In both cases, the central pulmonary arteries could be fully, and the segmental arteries partly, recanalized. Mobility of the system within the pulmonary arteries was limited. There was no evidence of wall damage at the rotation site of the impeller. Free serum hemoglobin did not increase after treatment. We conclude that the device can be safely operated in the left pulmonary artery system of dogs. It causes no significant hemolysis, and is able to accomplish rapid recanalization of the central arteries. Due to limited steerability, occluded side branches cannot be treated consistently. PMID- 8402788 TI - Stenosis of a surgical portosystemic shunt--treatment with angioplasty and placement of a Wallstent. AB - A case with a stenosed splenosuprarenal shunt 26 months after surgery is presented. Due to elastic recoil of the stenosis, percutaneous angioplasty was ineffective. However, stenting the stenosis with a Wallstent subsequently provided a satisfactory angiographic result and normal shunt function which is maintained at 4 months. PMID- 8402789 TI - Therapeutic embolization of intrahepatic portosystemic shunts by retrograde transcaval catheterization. AB - A 57-year-old woman presented with hepatic encephalopathy, cirrhosis, and a dual channel portosystemic venous shunt (PSVS). The shunt was treated successfully by embolization with steel coils via retrograde systemic venous access. Encephalopathy resolved. This new approach is considered safer than the previously reported percutaneous transhepatic route or the mesenteric venous route, requiring a mini-laparotomy. PMID- 8402790 TI - Balloon catheter dilatation of focal colonic strictures following necrotizing enterocolitis. AB - Two infants with severe colonic strictures secondary to necrotizing enterocolitis (NEC) were successfully dilated with balloon catheters. The procedures were performed under mild sedation, with fluoroscopic guidance. Results were sustained for more than 4 years follow-up. This procedure proved to be a safe and effective alternative to operative repair. PMID- 8402791 TI - An unopened titanium Greenfield IVC filter: intravascular ultrasound to reveal associated thrombus and aid in filter opening. AB - A titanium Greenfield filter did not open following placement in the infrarenal inferior vena cava (IVC). Abdominal radiograph and cavogram showed no definite reason for filter malfunction. Intravascular ultrasound (IVUS) demonstrated the unopened filter in the IVC with thrombus binding the legs. The thrombus was disrupted with a catheter, and the filter completely expanded with a balloon. IVUS documented full-filter opening in addition to residual thrombus in the filter following manipulation. PMID- 8402792 TI - Unsuccessful pericardiocentesis for cardiac tamponade during angiocardiography with nonionic contrast material. AB - Cardiac tamponade secondary to ventricular wall perforation is one of the possible complications of right-sided as well as left-sided cardiac catheterization. Ventriculography was performed on a patient with obstructive cardiomyopathy using a nonionic contrast medium. During the procedure, the right ventricle was accidentally perforated and the patient developed cardiac tamponade. Pericardiocentesis was unsuccessful and surgical drainage was needed after the patient developed progressive hemodynamic deterioration. At surgery, pericardial thrombus was found. We consider the reason for failure of pericardiocentesis was rapid coagulation of the extravasated blood in connection with nonionic contrast material. PMID- 8402793 TI - CT-guided large-bore biopsy: extrapleural injection of saline for safe transpleural access to pulmonary lesions. AB - A new technique was applied to improve safety for large-bore biopsies of subpleural pulmonary lesions. In 27 patients, normal saline solution (20-40 ml) was injected extrapleurally under CT-guidance in the access route to subpleurally situated lung lesions. Thickness of ventilated lung interposed between the target and the parietal pleural was up to 8 mm (mean: 4.1 mm). The injected fluid resulted in an extrapleural bulge that abutted the lesion to be biopsied. Biopsy with the 14-gauge Trucut needle did not result in pneumothorax in all cases. There were no complications associated with this technique. Extrapleural injection of saline may provide a safe access route to selected pulmonary lesions for CT-guided large-bore transthoracic biopsies. PMID- 8402794 TI - A simple and effective postangiographic femoral artery pressure dressing. AB - We describe a simple, inexpensive pressure dressing which supplements manual pressure and may be applied quickly and easily following femoral arterial puncture for angiography. This technique has been used on several thousand patients without complications. PMID- 8402795 TI - Mineral induction in vivo by dentine proteins. AB - The mineral-inductive capacity of polyanionic noncollagenous proteins, isolated from calcified tissues, has earlier been demonstrated in vitro. These macromolecules possess this capacity, provided that they are immobilized on a solid support, whereas in solution they inhibit crystal mineral induction and growth. The present study demonstrates this capacity also in vivo. Phosphoprotein, proteoglycan and gamma-carboxyglutamate-containing protein (Gla protein) of the osteocalcin type were covalently coupled to AH-Sepharose beads, and these were implanted subcutaneously in minipigs for periods between 3 and 24 weeks. Similarly, demineralized rat incisor dentine was implanted for 3 weeks, as was predentine dissected from dentinogenically active bovine incisors. The results were analyzed by scanning electron microscopy and energy dispersive X-ray analysis. Dentine proteoglycan and phosphoprotein coupled to beads were found to be inducers of calcium/phosphate-containing mineral, whereas the efficacy of Gla protein was negligible. Dissected predentine did not induce any mineral, but demineralized dentine preparations, still containing some noncollagenous anionic components, were found to induce mineral. The results indicate that macromolecules such as proteoglycan and phosphoprotein may be responsible for mineral induction during mineralization of calcified tissues. The results further imply that such molecules may be of importance for the remineralization of dentine in conjunction with the caries process. PMID- 8402796 TI - Detection and quantification of calcium fluoride using micro-Raman spectroscopy. AB - Investigations on the caries-preventive effect of calcium fluoride have been seriously hampered by the lack of adequate detection techniques. In this paper, the micro-Raman technique is introduced as a suitable method for CaF2 quantification with a spot size typically 5 microns. Advantages of this measuring technique and associated problems are described for CaF2 determination in the presence of large amounts of hydroxyapatite. The data show that a comparative analysis of Raman intensities of calcium fluoride and phosphate ions of hydroxyapatite can be used for quantification of CaF2 concentrations. The intensity behaviour of the CaF2 and hydroxyapatite Raman lines is explained considering the absorption-scattering cross-sections of the two components. The ratio of the Raman intensity (322 cm-1) of CaF2 to the phosphate Raman intensity (432/447 cm-1) of hydroxyapatite was found to be related linearly to the ratio of the concentration of CaF2 to that of hydroxyapatite. At present, the lower limit of CaF2 quantification is approximately 3 wt%. The technique has already been applied on fluoridated bovine enamel, and Raman emissions due to CaF2 or CaF2 like material formed on/in enamel have been detected for the first time. PMID- 8402797 TI - An in vitro evaluation of fluorescein penetration into natural root surface carious lesions. AB - In order to develop a method for detection and quantification of initial root surface carious lesions, the use of fluorescein sodium as a fluorescent dye is evaluated. The penetration depth of fluorescein sodium into human roots containing natural carious lesions was measured on approximately 130-microns thick slices, cut in a direction perpendicular to the pulp. This depth was compared to the lesion depth that was measured on the same slice with transversal microradiography. The results indicate that fluorescein sodium in water penetrates into demineralized layers of root dentine even if a surface layer with a high mineral content is present. The penetration depth is about 1.1 times the microradiographically determined lesion depth. PMID- 8402798 TI - Shrinkage prevention of in vitro demineralized human dentine in transversal microradiography. AB - The shrinkage of demineralized dentine is a serious problem in the assessment of mineral distributions and mineral changes by means of microradiography. In this article it is shown that a treatment with ethanediol of the thin slices used for transversal microradiography prevents shrinkage for about 1 h. Without this so called diol treatment, the shrinkage effects in demineralized dentin are about 20% in lesion depth and about 30% in mineral loss (delta Z) after 120-second drying in air. The prevention of shrinkage by diol treatment of dentine lesions is not influenced by the presence of a surface layer. PMID- 8402799 TI - Shrinkage of sound and demineralized human coronal dentine slabs. AB - In this article a method is presented for dentine shrinkage measurements. The relative shrinkage of sound dentine slabs is assessed using a Perthometer with respect to a steel reference. The relative shrinkage of lesions in dentine slabs can be estimated using combined Perthometer/microradiography measurements. The results show that the relative lesion shrinkage of lesions in thick slabs is quite different from the shrinkage of lesions in thin sections used for microradiography. The relative lesion shrinkage is, for periods up to 30 min, linear with the air drying period and increases strongly with the demineralization period. The previously published diol treatment is capable of reducing the relative lesion shrinkage in thick dentine slabs to negligible values. PMID- 8402800 TI - Distribution of fluoride in human dental calculus. AB - The distribution of fluoride in human dental calculus was investigated using a mechanical sampling technique and the fluoride electrode. The fluoride concentrations were highest at the outer surface of dental calculus and, then fell to a plateau for the interior of the calculus, rising again as the tooth surface was approached. The coefficient of variation of fluoride concentration was significantly greater in the sub- than in the supragingival calculus (p < 0.01). The supragingival calculus thus tended to show a smoother distribution profile for fluoride than the subgingival calculus. Total fluoride, average fluoride, and maximum fluoride concentrations were not significantly different in supra- and subgingival calculus. No significant different were observed between males and females. PMID- 8402801 TI - Control of calculus formation by a dentifrice containing calcium lactate. AB - The aim of this study was to re-investigate the anticalculus effects of calcium lactate. To this end calcium lactate was incorporated into experimental toothpastes. Three groups of 20 human volunteers brushed their teeth at least twice daily with three different toothpastes: (1) one containing calcium lactate, (2) one containing calcium lactate plus sodium lauryl sulphate, and (3) a control toothpaste. Plaque accumulation, gingival bleeding and calculus formation were scored at the beginning and at the end of the 3-month experimental period. After the first clinical examination the dentition was professionally cleaned to remove all dental deposits. The use of the calcium lactate containing toothpastes had no significant effects on plaque accumulation and gingival bleeding. However, calculus scores were significantly reduced in the calcium lactate groups at the end of the experiment. The results confirm and extend previous findings that the topical use of calcium lactate reduces calculus formation by, as yet, unexplained mechanisms. PMID- 8402802 TI - In situ anticariogenic potential of glass ionomer cement. AB - The purpose of this study was to compare the amount of fluoride in plaque formed on glass ionomer cement or composite and to evaluate the effects of fluoride released on growth of cariogenic microflora, fluoride uptake, and secondary caries formation under in situ conditions of a high cariogenic challenge. Ten adult volunteers took part in this crossover study performed in two phases of 28 days. Eighty enamel blocks were randomly restored with glass ionomer cement (Chelon-Fil-Espe) or composite (Silux). During each phase of the study, an acrylic resin appliance, containing four enamel blocks restored with the same material, was constructed for each of the volunteers. During the experimental period, all subjects used fluoride-free dentifrice, refrained from brushing the restored enamel blocks, and immersed the appliances into 20% sucrose solution eight times a day. Fluoride levels, mutans streptococci, and lactobacilli were assessed in dental plaque. Fluoride uptake and microhardness profiles were determined in enamel around the restorations. Statistical analyses indicated a significantly higher level of fluoride (p < 0.05) and a lower level of mutans streptococci plaque formed on glass ionomer cement. Analysis of variance in a split-plot model indicated that in the enamel around the glass ionomer restoration the fluoride uptake was significantly greater (p < 0.025) and the mineral loss significantly lower (p < 0.01). The results show that glass ionomer cement presents a broad anticariogenic effect and may be of value in preventing secondary caries, even under conditions of a high caries risk. PMID- 8402803 TI - Effects of a lactoperoxidase system-containing toothpaste on levels of hypothiocyanite and bacteria in saliva. AB - Lactoperoxidase (LPO)/H2O2/SCN(-)-system-generated hypothiocyanite ions (OSCN-) and hypothiocyanous acid (HOSCN) are inhibitory against a number of oral bacteria, including mutans streptococci. A commercially available toothpaste (Biotene) comprises the complete LPO system. Generation of HOSCN/OSCN- by Biotene was studied in vitro both in sterilized and nonsterilized saliva of 10 healthy subjects. The HOSCN/OSCN- yield ranged from 100 to 300 microM with Biotene, while the salivary levels of HOSCN/OSCN- before the addition of Biotene were 30.1 +/- 25.1 microM. Two in vivo trials were carried out. In the first study, resting saliva was collected from 12 individuals before, immediately after, and 2, 5, 10 and 20 min after brushing with Biotene to evaluate the in vivo generation and decomposition of HOSCN/OSCN-. In the second study, 26 healthy individuals attended a 1-month crossover trial with Biotene and a control toothpaste, Vademecum (no LPO system), both containing F- and xylitol. The salivary counts of total streptococci, mutans streptococci (MS), lactobacilli and the total flora (TF), as well as the peroxidase, thiocyanate ion and HOSCN/OSCN- levels were determined before and after 2 and 4 weeks daily use of the toothpastes. Twice-a day use of Biotene for 1 month resulted in an elevation of 'resting levels' of HOSCN/OSCN-. No such effect was found with the control toothpaste. No significant changes were found in the salivary levels of total streptococci, MS, lactobacilli or TF after 1-month use of either toothpaste. The results show the capability of the LPO-system-containing toothpaste to elevate the salivary levels of HOSCN/OSCN , although no bactericidal effect was observed. PMID- 8402804 TI - Close association between Streptococcus sobrinus in the saliva of young children and smooth-surface caries increment. AB - Three hundred and thirty-eight children (age range 3-5 years) were examined for the presence of mutans streptococci in their saliva. They were divided into four groups according to the prevalent species that were isolated: (1) Streptococcus mutans and Streptococcus sobrinus group (S. mutans > or = 10(3) CFU/ml and S. sobrinus > or = 10(3) CFU/ml); (2) S. mutans group (S. mutans > or = 10(3) CFU/ml but S. sobrinus < 10(3) CFU/ml); (3) S. sobrinus group (S. sobrinus > or = 10(3) CFU/ml but S. mutans < 10(3) CFU/ml), and (4) children with mutants streptococci absent or barely detectable (mutants streptococci < 10(3) CFU/ml). The previous caries experience was compared among the groups, as was the caries increment during a 6-month period after the bacterial examination. The subjects in the S. sobrinus group had both higher caries and higher decayed teeth and surface increments than the S. mutans group. The mean decayed smooth-surface increment (delta d-surface) for the S. sobrinus group was 2.6 and was significantly higher than 0.8 for the S. mutans group. For the subjects with delta d-surface > or = 4, the mean number of S. sobrinus in the saliva was 4.29 x 10(5) CFU/ml and was significantly higher than 0.32 x 10(5) CFU/ml for the subjects without delta d surface. There was no significant difference in the mean number of S. mutants in saliva between the subjects with and without delta d-surface. The prevalence of S. sobrinus in saliva was more closely associated with future caries activity, especially with smooth-surface caries increment, than the prevalence of S. mutants. PMID- 8402805 TI - The effect of chlorhexidine and zinc/triclosan mouthrinses on the production of acids in dental plaque. AB - Chlorhexidine, and zinc in combination with triclosan, are used as anti-plaque agents in the prevention of gingivitis. The multifunctional activity of these compounds against bacterial cells has been proposed to include interference with sugar transport and reduction of glycolysis. In this study the ability of the agents to reduce acid production in dental plaque in vivo has been investigated. Samples of smooth-surface plaque were collected from individuals who had been rinsing for several weeks with (a) chlorhexidine (0.12%, Peridex); a combination of zinc and triclosan in mouthwashes containing (b) high and (c) low concentrations of humectant; or (d) a control mouthwash. Analyses using isotachophoresis showed that resting plaques in the chlorhexidine and zinc/triclosan groups contained less acetate than the control group. Acids were also measured 15 min after a glucose rinse. Compared with the control, the amount of lactate was significantly decreased (45%) in the chlorhexidine group, while lactate reduction (20%) in the zinc/triclosan (high humectant) group was not statistically significant. PMID- 8402806 TI - Effect of single and repeated application of chlorhexidine varnish on mutans streptococci in plaque from fissures of premolar and molar teeth. AB - The aim of this study was to determine the effects of one and two applications of 40% chlorhexidine varnish on the numbers of mutants streptococci in human dental fissure plaques from molar and premolar teeth. Twenty-nine subjects (aged 20-30 years) participated in the study and were randomly assigned to one of three groups. In each subject two fissures with high levels of mutants streptococci were selected. The fissures in group 1 (control group) were treated with placebo varnish containing no chlorhexidine. Fissures in group 2 received a single application of 40% chlorhexidine varnish, whereas fissures in group 3 received an additional chlorhexidine varnish application 1 week after the first application. Fissure plaque samples were taken prior to the first application of chlorhexidine varnish and subsequently 1, 2, and 4 months thereafter. Compared with the fissures in the control group, the suppression of mutans streptococci was significant in plaques from group 2 for up to 2 months and in plaques from group 3 for up to 4 months after application. Mutans streptococci were suppressed more strongly in premolar than in molar teeth and more strongly and for a longer period of time in fissures of premolar teeth treated twice than in fissures of premolars treated once. PMID- 8402807 TI - Validity and reproducibility of clinical examination, fibre-optic transillumination, and bite-wing radiology for the diagnosis of small approximal carious lesions: an in vitro study. AB - This study measured validity and reproducibility of unaided clinical diagnosis, fibre-optic transillumination (FOTI), and bite-wing radiology in the diagnosis of approximal caries. Sixty models were made using extracted premolars and molars, each containing four teeth with six contacting approximal surfaces. The teeth were examined first using unaided clinical examination and then using FOTI. Bite wing radiographs were then taken of the teeth set in the models and examined. The three examinations were repeated after 1 week. Histological sections of the undecalcified teeth were prepared following their removal from the models, and those showing signs of caries were examined to give the valid state of disease in each surface. The diagnostic threshold was caries penetrating into dentine. The reproducibility of all three methods was acceptable with kappa values exceeding 0.6. All specificity values exceeded 0.95. Statistically significant differences were seen between sensitivities for clinical (0.38) and bite-wing (0.59) diagnosis and between clinical and FOTI (0.67) examination, but not between bite wing and FOTI. It is concluded that the validity of FOTI is at least as high as that of bite-wing radiology, and both are superior to unaided clinical diagnosis. PMID- 8402808 TI - Validity of bite-wings for diagnosis of secondary caries in teeth with occlusal amalgam restorations in vitro. AB - This investigation was carried out to establish the validity of bite-wing radiographs for the diagnosis of secondary caries in teeth with occlusal amalgam restorations. One hundred and fifty-nine extracted molars with occlusal amalgam fillings and characteristics that might be indicative of the presence of secondary caries were selected. The characteristics were a blueish-gray discolouration of enamel, a brownish discolouration of the amalgam-enamel margin, marginal breakdown of the restoration, and/or deep fissures at the margin of the restoration. The teeth were radiographed in a bucco-lingual direction to obtain images comparable to bite-wings. Subsequently, the crowns were sectioned (700 microns) and glued on plastic sheets. The sections were recorded radiographically and then examined by independent examiners. Radiolucencies as well as radiopacities adjacent to the restoration were considered to be caries. The validation of the bite-wing radiographic diagnosis was achieved by comparison with the radiographs of the sections. A sensitivity of 64% and a specificity of 98% were calculated. Large lesions were always visible on the bite-wing radiograph (100%), the majority of medium-sized lesions (89%) and 40% of small lesions. Small radiopacities were detected more often than small radiolucencies. PMID- 8402809 TI - Enamel defects in 4- to 5-year-old children in fluoridated and non-fluoridated parts of Cheshire, UK. AB - The aim of this study was to compare the prevalence of developmental defects of enamel in the deciduous dentition of 4- to 5-year-old children residing in fluoridated (1 ppmF) and non-fluoridated (less than 0.2 ppmF) communities in Cheshire, UK. The significant difference in the prevalence of developmental defects of enamel between the two communities was accounted for by the higher prevalence of diffuse opacities in fluoridated Nantwich (29%), than in non fluoridated Northwich (14%). The results also showed that when controlling for the age at which parents claimed toothbrushing commenced, the children in fluoridated Nantwich still had significantly more diffuse defects than the children in Northwich. PMID- 8402810 TI - Methods of projecting long-term relative efficacy of products exhibiting small short-term efficacy. AB - There is ample evidence that caries can develop well into the middle-aged years, and for many subjects, for most of their adult lives. The kinds of benefits to expect for adults in the longer term for improved fluoride products, especially dentifrices, based upon small observed relative improvements in short-term efficacy is a topic currently of considerable interest. For example, what longer term benefits can be expected by the continued use of an improved fluoride dentifrice, corresponding to an observed 5% or 10% short-term improved efficacy relative to a positive control product? In this study, three models are presented and discussed for projecting longer-term relative benefits of fluoride products for which valid short-term efficacy estimates are available: the compound growth model (CGM), which is a derivative of the compound interest model; the demineralization square root model (DSR), used in demineralization remineralization studies, and the stabilization model (STA), which is based on the assumption of constant caries attack rates. Published results from long-term studies of fluoride dentifrices and other topical fluoride products (rinses and tablets) suggest that anticaries benefits for topical fluorides do propagate over time, providing increased relative benefits to patients in the longer-term, as compared with the benefit estimates from short-term clinical experiences.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402811 TI - Comparative efficacy of NaF and SMFP dentifrices in caries prevention: a meta analytic overview. AB - An overview was conducted of all randomized, controlled studies comparing NaF to SMFP dentifrices in the prevention of caries development. Data from these separate trials were subjected to a pooling procedure, or meta-analysis, in order to obtain a more stable estimate of comparative treatment efficacy and to aid in interpreting the generalizability of results. Based on a pool of studies involving over 7,000 subjects, NaF was associated with a significantly greater reduction in caries development compared to SMFP. The increment in D(M)FS was reduced an average of 0.28 (95% confidence limits 0.10 to 0.46) with the use of NaF as compared to SMFP over a 2- to 3-year follow-up period. This difference represents a 6.4% reduction in the rate of caries development observed with SMFP. The numerical advantage conferred by NaF over SMFP in caries risk reduction must be judged clinically as to its public health implications. Alternative analytic techniques and rules for including studies in the pooling process yielded consistent conclusions. A similar analysis of data from dual-active studies indicated that NaF in combination with SMFP provides greater lowering of the D(M)FS increment (approx. 0.16 over 2-3 years) compared to SMFP at the same total fluoride dose, whereas the dual-active product demonstrated no advantage over NaF alone. PMID- 8402812 TI - A critical review of the relative anticaries efficacy of sodium fluoride and sodium monofluorophosphate dentifrices. AB - While there is broad consensus in the research community that fluoride dentifrices provide important anticaries benefits, debate still remains as to the most efficient form of fluoride used in toothpastes. Recently, the authors of this paper collaborated as part of a scientific advisory group whose objective was to comprehensively review all clinical information available comparing the anticaries efficacy of the two agents most widely used in fluoridated toothpastes, sodium fluoride (NaF) and sodium monofluorophosphate (SMFP). This review included a detailed analysis of each published study pertinent to the question, a comprehensive meta-analysis of all available clinical findings, and an epidemiological assessment of how anticaries benefits of dentifrices may be anticipated to propagate with time. Overall, the use of meta-analysis of head-to head clinical comparisons between the two fluoride-active systems was found to be the most valid means for comparing the relative efficacy of NaF and SMFP dentifrices. Results of this analysis demonstrated that NaF was significantly more effective than SMFP in preventing caries (p < 0.01). While the numerical difference in efficacy between NaF and SMFP measured between 5 and 10% (total DMFS) over a 2- to 3-year clinical period, this could be expected, on theoretical grounds, to propagate to substantially larger differences (e.g. 10-20%) over 10 20 years. Hence, the difference in efficacy between these two actives was judged to be clinically important as well as statistically significant. Based upon these findings the authors recommend that NaF be used as the active system in fluoridated dentifrices whenever practically feasible. However, the authors caution that this recommendation pertains to the formulation of NaF in highly compatible abrasive systems, which must be demonstrated by critical evaluation of ionic fluoride within formulations for stability, availability and bioavailability. One additional recommendation emanating from this review is that important improvements must be made in the design, execution and reporting of future caries clinical trials in order to bring these important methods up to scientifically acceptable standards. PMID- 8402813 TI - [Congenital heart defects in adults from the viewpoint of the surgeon]. AB - The adolescent generation of patients operated in childhood on account of congenital heart disease is a new health and social problem. Kardiocentrum for children in Motol started its activities in 1977. A concept of care for children with congenital heart disease was elaborated and gradually all surgical methods used worldwide were introduced. The advances in cardiosurgery along with the development of diagnostic methods and intervention catheterization in particular balloon valvuloplasty and angioplasty make high standard and effective treatment of practically the entire spectrum of congenital heart disease in children of all age groups possible. The cardiac surgeon tries nowadays to resolve problems of inborn heart disease already in early childhood. Before a possible re-operation in adult age he must take into account how the result of the first operation affected the patient's growth and development. He is interested in particular in the fate of implants, surgically induced pathology and the influence of growth on the reconstruction. Despite intensive research, so far the long-term prognosis of patients after some types of surgery is not clear (e.g. transposition of the great arteries, Fontan's operation and its modifications). An adult patient congenital hearth disease should receive care of a specialized cardiologist who has personal experience with the diagnosis and treatment of congenital heart disease. If surgery is necessary it should be performed in a specialised department equipped as regards staff, technical devices and funds for these special operations. PMID- 8402814 TI - [The Prague Diabetes Registry. 3. Retinopathies, nephropathies and neuropathies in type 1 diabetics. The Prague Diabetes Collective]. AB - In the Prague register of all adult diabetics by April 1, 1992 data on 1443 patients above 18 years with type 1 DM were recorded. In 42.2% of diabetics diabetic retinopathy was observed which is manifested most frequently after 11 to 15 years' persistence of diabetes. Proteinuria, the first sign of diabetic nephropathy, was found in 13.8% of the group, 5.4% of the diabetics are in the stage of renal insufficiency. Nephropathy is 3 to 4 times more frequent in diabetics with retinopathy. Peripheral neuropathy is found in 32.8% type 1 diabetics. PMID- 8402815 TI - [Tick-borne encephalitis in the West Bohemian Region 1960-1991]. AB - In 1960-1991 in the West Bohemian region 746 cases of tick-borne encephalitis were confirmed by serological examination. The highest morbidity was recorded in the population of Plzen--5.9 per 100,000 per year. During 1986-1991 the mean hospitalization period of patients with tick-borne encephalitis was 13.5 days. Analysis by age groups revealed the highest specific morbidity in subjects aged 15-34 years. In the limital age groups the disease was recorded only exceptionally. From data on the specific morbidity and clinical severity ensues that mass vaccination against this infection in areas with a lower risk could start in subjects older than 10 years. PMID- 8402816 TI - [Occurrence of acute infectious diarrhea during the lunar phases]. AB - A chronobiometric analysis of 753 cases of acute infectious diarrhoea in adults in 1981-1990 in Kosice confirmed to a surprising extent recently reached conclusions of an investigation made by authors from Bratislava. The Kosice group comprised 352 cases of bacillary dysentery, 305 patients with salmonellosis, 72 with campylobacteriosis and 24 with yersiniosis. Statistically significantly fewer patients (p < 0.0001) were hospitalized during full moon, moon quarterly and new moon. In the intervals there were periods with a short-term increase of the daily admissions by cca 25%. This 7.38-day periodicity cannot be explained by the influence of the social 7-day week, as during observations extending over several years this rhythm is eliminated by a gradual shift across different phases of the moon. The authors did not find similar reports in the literature. For explanation, not only the organism of the host (variable immunity?) but also the infectious agent must be taken into account. More profound understanding of the mechanism may open the road to practical application of the described lunar relationship. Its knowledge can help already now to improve the organization of the health service. PMID- 8402817 TI - [Trimethoprim in the therapy of symptomatic liver porphyria]. AB - Administration of antimalaria drugs to patients with symptomatic hepatic porphyria (porphyria cutanea tarda) at the First Medical Clinic made it possible due to previous experimental studies, to influence the porphyrim metabolism of yeasts. The effect of trimetoprim was investigated in clinical work in 12 hitherto not treated patients with the manifest form of symptomatic hepatic porphyria. The group comprised 9 men, mean age 56.4 years, and 3 women mean age 43.6 years. During the two-year clinical study dermatological symptoms of the disease became milder or receded. Concurrently there was a significant regression of porphyrinuria. In the whole group porphyrinuria declined to 11% of the original values. The porphyrin content of hepatic tissue declined considerably after two years treatment. In the group as a whole to 40% of the original values. Trimetoprim is another drug which influences porphyrin metabolism. The authors did not detect any undesirable side-effects of trimetoprim treatment. The effect of trimetoprim on clinical and biochemical parameters of the disease is less marked than the effect of chloroquine. This new treatment can be used in patients resistant to chloroquine or in combination with other anti-malaria drugs. PMID- 8402818 TI - [Physicians and the Sokol]. PMID- 8402819 TI - [Non-cholinergic effects of organophosphates]. AB - The author summarizes contemporary knowledge on the action of organophosphates with emphasis on non-cholinergic mechanisms. Intoxication with these substances is characterized by four basic processes: absorption, transport, metabolism and the toxic effect proper. The latter involves cholinesterase inhibition at the cholinergic synapses with subsequent accumulation of acetylcholine. However, non cholinergic processes are also involved such as the effect on levels of cyclic nucleotides, the peptidergic system, receptors, liver metabolism and the stressogenic action. From the review ensues that knowledge of non-cholinergic, mechanisms of action could reveal new approaches to treatment of intoxications with these substances. PMID- 8402820 TI - [Changes in body fluids and minerals after cold adaptation]. AB - To characterize fluid and ion shifts during 5-weeks cold adaptation, 6 nonadapted volunteers underwent cold acclimation programme (CAP), consisted of 1 h head-out immersion in water 14 +/- 1 degrees C 3 times a week. Blood samples were analyzed before/after the immersion and in the first and in the last week of CAP. Urine was collected for 10 h before, during, and after immersion for 4 h. Plasma volume (PV) decreased during first immersion (-18%) and less after CAP (-12). Blood volume reduction was 8.5% before and 5.2% after the CAP. Mean corpuscular volume was not changed either after the cold exposition or after the cold adaptation. The concentration of serum proteins increased by 12.1% after first immersion and by 8.1% after the CAP. The changes in serum concentration of Na+, K+ and Cl- before and after the CAP were not significant. Urine flow increased by 102% after first immersion, and by 165% after CAP. Urinary excretion of Na+ increased by 167% and 283%, excretion of K+ by 222% and 362% during first immersion and after CAP, respectively. Serum concentration of aldosterone increased nonsignificantly (+30%) during immersion before CAP and it did not changed after CAP. After the cold adaptation we observed the reduction of PV decrease, and increased diuresis with higher excretions of cations. PMID- 8402821 TI - [Comparison of immunochemical and biochemical tests for occult hemorrhage in feces]. AB - The authors compared in a group of 104 patients assessment of occult haemorrhage in faeces made by the immunochemical test HemeSelect (HS-SmithKline Diagnostics) and the biochemical Haemoccult test (HT-Rohm Pharma). In all patients subsequently total coloscopy was performed. In the whole group the sensitivity of HS was 49.3% and of HT 16.0% (p << 0.001) and the specificity 72.4% and 100% (p << 0.01). The predictive value in polyps, 10 mm in size or larger, was 65.2% (positive) and 60.0% (negative). The assessed sensitivity and predictive values of HS do not permit to recommend immunochemical tests of this type as an alternative procedure to repeated colonoscopic checks in the dispensarization of high risk groups of colorectal cancer. PMID- 8402822 TI - [Immunologic follow-up of HIV-positive patients. C3, C4 complement components during HIV infection]. AB - Investigations of the authors' patients with HIV infection revealed marked changes in the C3 and C4 components of complement. Of 61 investigated patients with HIV infection only 23 patients (37.7%) had values of the C3 component at the level recorded in controls. Values of the C4 component comparable with the control group were recorded in 24 patients (39.3%). From figures 1 and 2 it apparent that in the investigated group there is a striking hypercomplementaemia of both components, as compared with hypocomplementaemia. PMID- 8402823 TI - [Systemic administration of antibiotics in therapy of brain abscesses. Present trends and possibilities]. AB - The authors describe possibilities as regards selection of antibiotics in the general treatment of brain abscesses. They mention the most frequent aetiological agents of the disease in relation to predisposing factors and site. Systemic antibiotherapy along with surgery is considered by the authors decisive in the therapy of these conditions. PMID- 8402824 TI - [The first hyperimmune immunoglobulin against cytomegaloviruses made in the Czech Republic]. AB - The Czech immunoglobulin CMV was administered to 16 subjects with active CMV infection--mostly women with repeated miscarriages, CMV primoinfection during pregnancy, to children with congenital CMV infection and to patients with other clinical forms. The immunoglobulin was administered in a single dose or repeatedly by the intramuscular route. The decline of activity of the CMV infection was demonstrated by a decline of specific IgM CMV antibodies, disappearance of previously assessed viriuria and isolation of CMV in smears from the uterine cervix. PMID- 8402825 TI - Light- and electron-microscopic immunocytochemistry of somatotropes in the anterior pituitary gland of European ferret, Mustela putorius furo. AB - Light-microscopic immunocytochemistry of ferret anterior pituitary revealed the localization of somatotropes in the pars distalis, but no immunoreactive cells were detected in the pars tuberalis. Ultrastructural studies by superimposition immunocytochemistry and immuno-electron microscopy, elucidated the morphological heterogeneity of these somatotropic cells. They were classified into 2 subtypes on the basis of size of the secretory granules. Type-I cells with small granules (mean diameter, 192 nm), were considered to be the immature somatotrop, while Type-II cells, with comparatively larger secretory granules (mean diameter, 257 nm), were considered to be the matured form of Type-I cells and the typical somatotropic cell-type, and were much more predominant than the Type-I cells. The fact that Type-II cells had a distinct Golgi zone and many mitochondria, while in Type-I cells the intracellular organelles were generally less developed, supports this suggestion. In addition to these two extreme subtypes, several intermediate forms were also encountered that may represent different transitional phases during the conversion of Type I to Type II. Protein A-gold immuno-electron microscopy illustrated the specific localization of growth hormone over the granules, with no labelling over any other cytoplasmic organelles of the 2 somatotrope subtypes. PMID- 8402826 TI - A new type of putative non-visual photoreceptors in the optic lobe of beetles. AB - A putative photoreceptor organ is described in the carabid beetle, Pachymorpha sexguttata. The elongated structure, about 20-40 microns wide and more than 300 microns long, is situated within the optic lobe at the fronto-dorsal rim of the lamina. It lies, deep in the head capsule, in front of the compound eyes and beneath window-like thinnings of the cuticle. The organ is composed of two types of cells: (1) clear sheath cells and (2) well-organized inner receptor cells that appear in a horseshoe-like or circular array in cross-section. Common histological features of all inner cells include a distal trunk ending in microvilli that form a rhabdom-like structure, an axon at the proximal end of the cell, lamellar and multivesicular bodies within the trunk, and clusters of small mitochondria. The organ has no shielding pigment. It is connected by thin axons to a circumscribed neuropil that parallels the organ, and thence via a fiber tract to the medulla accessoria, a possible site of the circadian pacemaker in insects. Immunoreactivity to anti-per(s), an antibody recognizing the Drosophila period (per) protein that plays a central role in the function of the circadian pacemaker in fruit flies, is demonstratable in thin efferent terminals within the organ, in the associated neuropil and in its fiber connection to the medulla. A second receptor organ displaying the same fine structure lies near the second optic chiasm. This set of putative photoreceptors also occurs in the tenebrionid beetle, Zophobas morio, and its pupa. The possible function of these receptor organs is discussed with respect to former chronobiological data and some recently described types of extraretinal photoreceptors in arthropods. PMID- 8402827 TI - Developmental changes in intestinal globule leukocytes of normal rats. AB - The changes in the number, distribution, and ultrastructure of globule leukocytes (GL) during postnatal development were investigated in the intestinal epithelium of non-infected healthy rats. Intestinal GL were abundant in normal newborn rats even in the absence of infection. They subsequently decreased markedly to the adult level by the fourth week. Ultrastructurally, morphological variations suggesting maturation of the cells were observed in the GL during development. These changes could be noted neither in the mucosal mast cells (MMC) nor in the granular intra-epithelial lymphocytes. Morphological differences between GL and other cells were evident in adult animals. Most notably, paracrystalline structures were found exclusively in the granules of the GL. Immunohistochemically, both the GL and MMC were stained with anti-serotonin, but not with anti-IgE. Degranulation of GL in developing rats was caused by repeated intraperitoneal administration of dexamethasone. Neither GL nor MMC were affected by compound 48/80. These results indicate that (1) the GL and MMC are derived from a common ancestral cell toward the end of embryonic development, (2) the immature GL migrate from the lamina propria into the epithelium to differentiate, mature, and proliferate, and (3) the immature GL have specific functions during the neonatal period. PMID- 8402828 TI - Relationship between the Golgi complex and microtubules enriched in detyrosinated or acetylated alpha-tubulin: studies on cells recovering from nocodazole and cells in the terminal phase of cytokinesis. AB - Double immunofluorescence microscopy was used to study the relationship between the Golgi complex and microtubules enriched in posttranslationally modified tubulins in cultured mouse L929 fibroblasts. In interphase cells, the elements of the Golgi complex were grouped around the microtubule-organizing center. From here, tyrosinated microtubules extended to the periphery of the cells, whereas the distribution of detyrosinated and acetylated microtubules largely overlapped with that of the Golgi complex. Treatment of cells with 10 microM nocodazole led to the disruption of all microtubules and dispersion of the Golgi elements. Following withdrawal of the drug, tyrosinated microtubules reformed first, followed by acetylated and then detyrosinated microtubules. In parallel, the Golgi elements moved back toward the juxtanuclear region and reestablished a close spatial relationship first with the acetylated and later also with the detyrosinated microtubules. Long-term recovery in the presence of 0.15 or 0.3 microM nocodazole allowed partial reformation of tyrosinated and acetylated microtubules, whereas no or only a few detyrosinated microtubules were detected. At the same time, the Golgi elements were grouped closer together around or on one side of the nucleus in close relation to acetylated microtubules. In synchronized cells released from a mitotic block, a radiating array of tyrosinated microtubules was first formed, followed by acetylated and detyrosinated microtubules. The Golgi elements initially came together in a few groups and thereafter took an overall morphology similar to that in interphase cells. During this reunification, they showed a close spatial relationship to acetylated microtubules, whereas detyrosinated microtubules appeared only later. Microtubules enriched in acetylated and/or detyrosinated tubulin thus appear to take part in establishing and maintaining the organization of the Golgi elements within an interconnected supraorganellar system. Whether the acetylation and detyrosination of tubulin are directly involved in this process or merely represent two modifications within this subpopulation of microtubules remains unknown. PMID- 8402829 TI - Basic fibroblast growth factor in rat salivary glands. AB - We studied the occurrence and localization of basic fibroblast growth factor (bFGF) in rat salivary glands using a specific monoclonal antibody. It was shown that the extract of rat salivary glands has a pronounced stimulatory activity on the growth of bovine capillary endothelial cells, which is blocked by the addition of an antibody against bFGF. The concentration of bFGF in the submandibular/sublingual gland, as determined by radioimmunoassay, was approximately 80% that in the brain. Immunocytochemistry revealed bFGF immunoreactivity localized primarily in the epithelial cells lining the striated ducts and excretory ducts of the parotid, sublingual and submandibular glands. In addition, intense bFGF-immunoreactivity was observed in the granular convoluted tubule of the submandibular gland, localized predominantly in the agranular pillar cells, which lay in small numbers among the majority of weakly immunostained cells containing many apical secretory granules. At the electron microscopic level, the immunoreactive material was distributed diffusely in the cytoplasmic matrix and nuclei of all immunoreactive cells, whereas it was absent from all cytoplasmic organelles including the secretory granules. These results indicate that bFGF is localized in different cellular and subcellular compartments from those of other growth factors in the duct system of rat salivary glands. PMID- 8402830 TI - Developmental changes at the node and paranode in human sural nerves: morphometric and fine-structural evaluation. AB - Developmental alterations of paranodal fiber segments have not been investigated systematically in human nerve fibers at the light- and electron-microscopic level. We have therefore analyzed developmental changes in the fine structure of the paranode in 43 human sural nerves during the axonal growth period up to 5 years of age, and during the subsequent myelin development up to 20 years and thereafter. The nodal, internodal, and paranodal axon diameters reach their adult values at 4-5 years of age. The ratio between internodal and paranodal axon diameters remains constant at 1.8-2.0. Despite a considerable increase in myelin sheath thickness, the length of the paranodal myelin sheath attachment zone at the axon does not increase correspondingly, because of attenuation, separation from the axolemma, and piling up of myelin loops in the paranode. Separation of variable numbers of terminal myelin loops from the underlying axolemma results in the formation of bracelets of Nageotte, whereas the transverse bands of these loops disappear. The adaptation of the paranodal myelin sheath to axonal expansion during development probably occurs by uneven gliding of the paranodal myelin loops simultaneously with internodal slippage of myelin lamellae. Since mechanically stabilizing structures (tight junctions and desmosomes between adjacent paranodal myelin processes; transverse bands between myelin loops and paranodal axolemma) are unevenly arranged, especially during rapid axonal growth, paranodal axonal growth with simultaneous adaptation of the myelin sheath is probably discontinuous with time. PMID- 8402831 TI - Collagen fibrillogenesis in a three-dimensional fibroblast cell culture system. AB - The purpose of this study was to follow collagen fibril formation in a newly developed three dimensional cell culture system. Human neonatal foreskin fibroblasts were grown on a nylon mesh in Dulbecco's Modified Eagles Medium (DMEM) supplemented with 10% fetal calf serum and antibiotics. Fibrillogenesis was initiated by the addition of 50 micrograms/ml ascorbate to confluent cultures. Sample meshes were processed for electron microscopy or immuno-electron microscopy. Fibrils approximately 20-30 nm in diameter, with 67 nm periodicity, were first detected five days after the addition of ascorbate. As cultures progressed, cells organized into parallel layers between which collagen fibers continued to form and increase in diameter. By day 50, fiber diameter ranged from 30 to 80 nm and large bundles were seen. No collagen fibril formation occurred in control cultures to which no ascorbate was added. However, large amounts of microfibrils were observed. Antibodies against the aminopropeptide of type I procollagen were found to bind to fibrils with diameters less than 34 nm while antibodies against the aminopropeptide of type III collagen bound primarily to fibers which ranged from 35-54 nm in diameter. We believe that this system, which morphologically resembles a normal dermis, will serve as an excellent model for the study of collagen fibrillogenesis. PMID- 8402832 TI - Reticulum cells in the ontogeny of nasal-associated lymphoid tissue (NALT) in the rat. AB - This study concerns the ontogeny of reticulum cells (RC) in the nasal-associated lymphoid tissue (NALT) of Wistar and Brown-Norway rats. A panel of monoclonal antibodies (mAb) directed against RC in peripheral lymphoid organs (antibodies ED10-ED15) was used, together with a recently developed antibody ED17, which recognizes macrophages and Langerhans cells. Early in embryogenesis, staining with common connective tissue markers, ED14 and ED15, was found. ED17-positive cells were present before cells positive to ED1, a pan-macrophage marker, or Ia glycoproteins were observed. The first differentiation of reticulum was seen at the day of birth, when ED10 recognized a distinct area in the nasal mucosa. The first T-lymphocytes were found at the same time. Two days after birth, B-cells and ED11-positive cells were present in the NALT area. Fourteen days after birth, T- and B-cell compartments were recognizable. ED10 was found predominantly in the T-cell area and ED11 was mainly confined to the B-cell compartment. We conclude that the development of the NALT is closely accompanied by the phenotypic specialization of the reticulum. This suggests that the reticulum plays an important role in the compartmentalization of NALT tissue and in the retention of lymphocyte subsets within these compartments. PMID- 8402833 TI - The embryonic development of the Drosophila visual system. AB - We have used electron-microscopic studies, bromodeoxyuridine (BrdU) incorporation and antibody labeling to characterize the development of the Drosophila larval photoreceptor (or Bolwig's) organ and the optic lobe, and have investigated the role of Notch in the development of both. The optic lobe and Bolwig's organ develop by invagination from the posterior procephalic region. After cells in this region undergo four postblastoderm divisions, a total of approximately 85 cells invaginate. The optic lobe invagination loses contact with the outer surface of the embryo and forms an epithelial vesicle attached to the brain. Bolwig's organ arises from the ventralmost portion of the optic lobe invagination, but does not become incorporated in the optic lobe; instead, its 12 cells remain in the head epidermis until late in embryogenesis when they move in conjunction with head involution to reach their final position alongside the pharynx. Early, before head involution, the cells of Bolwig's organ form a superficial group of 7 cells arranged in a 'rosette' pattern and a deep group of 5 cells. Later, all neurons move out of the surface epithelium. Unlike adult photoreceptors, they do not form rhabdomeres; instead, they produce multiple, branched processes, which presumably carry the photopigment. Notch is essential for two aspects of the early development of the visual system. First, it delimits the number of cells incorporated into Bolwig's organ. Second, it is required for the maintenance of the epithelial character of the optic lobe cells during and after its invagination. PMID- 8402834 TI - Current bibliography of cell calcium prepared by University of Sheffield Biomedical Information Service. PMID- 8402835 TI - Effects of elevated pressures of inert gases on cytosolic free Ca2+ of human platelets stimulated with ADP. AB - Platelets were exposed to elevated pressures of helium (He), nitrogen (N2), and He/xenon (He/Xe, 85/15), and stimulated with ADP (5 microM). He to 36 atmospheres absolute (ATA) inhibited the ADP-stimulated increase in [Ca2+]i, measured by Fura 2/AM, with most of the effect occurring by 9 ATA. N2 caused a similar but initially greater effect and it was bimodal, with inhibition being less at 36 ATA than at 18 ATA. N2 also significantly depressed basal levels at 18 and 36 ATA. In the He/Xe mixture, the effect on ADP-stimulated [Ca2+]i was no different from He alone at the same pressures, but basal levels were significantly depressed. In a calcium-free medium both He and N2 moderately depressed the already-reduced response to ADP stimulation but only N2 significantly depressed basal levels at all pressures. These results indicate that raised pressures of inert narcotic gases, as well as pressure per se (represented by He), can affect cell [Ca2+]i sufficiently to have physiological consequences. This, together with previous findings in this laboratory, may have implications for some of the neurological problems associated with deep diving. PMID- 8402836 TI - Characterization of the inositol trisphosphate-sensitive and insensitive calcium stores by selective inhibition of the endoplasmic reticulum-type calcium pump isoforms in isolated platelet membrane vesicles. AB - In mixed platelet membrane vesicles the presence of two distinct endoplasmic reticulum-type calcium pump enzymes of 100 and 97 kD molecular mass has been demonstrated. We have previously shown that both calcium pumps were recognized by polyclonal anti-sarcoplasmic reticulum calcium pump antisera [11]. In the present work we studied the effects of several calcium pump inhibitors on active calcium transport and inositol trisphosphate-induced calcium release in these vesicles in an attempt to assign the two calcium pump isoenzymes to specific calcium pools. The effect of the PL/IM 430 inhibitory anti-calcium pump antibody was compared to that of other calcium pump inhibitors acting predominantly on the 100 and the 97 kD calcium pump isoforms, respectively. The PL/IM 430 antibody, which recognized the 97 kD pump on Western blots and 2,5-di-(tert-butyl)-1,4-benzohydroquinone, which inhibited phosphoenzyme formation of the same pump isoform, inhibited calcium accumulation predominantly into an inositol trisphosphate-releasable calcium pool. On the other hand, low concentration of thapsigargin, which inhibited phosphoenzyme formation mainly of the 100 kD pump isozyme, had a more pronounced effect on calcium uptake into an inositol trisphosphate-resistant pool. These data suggest that in platelets the 97 kD calcium pump isoform is likely to be associated with the inositol trisphosphate-sensitive calcium storage organelle. PMID- 8402837 TI - Altered calcium regulation in SV40-transformed Swiss 3T3 fibroblasts. AB - Calcium homeostasis has long been thought to be altered in transformed cells but mechanisms have not been established. In this study, the photoprotein, aequorin, was used to examine calcium regulation in 3T3 and SV40-transformed 3T3 cells. It was found that calcium transients induced by bradykinin or serum in serum-starved cells are lower and delayed in the transformed cells and decay kinetics are altered. These changes are not related to differences in cell cycle distribution. Though the serum transient is insensitive to nifedipine, verapamil, or lanthanum, removal of extracellular calcium accelerates transient decay in both cell types. Treatment of unstimulated cells with the ER Ca(2+)-ATPase inhibitor, thapsigargin, causes a 4-5-fold greater increase in [Ca2+]i in the transformed than in the nontransformed cells. Following serum stimulation, transformed cells still exhibit a large thapsigargin-induced increase in [Ca2+]i whereas the response in nontransformed cells is nearly abolished. When the 3T3 or SV3T3 cells are exposed to serum or thapsigargin in the absence of extracellular calcium and subsequently exposed to 11.8 mM Ca2+, a much greater influx of calcium again occurs in the SV3T3 cells. The observed changes in SV3T3 cells are most likely due to an alteration in a capacitative mechanism which regulates influx of calcium through the plasma membrane. PMID- 8402838 TI - Changes in cytosolic resting ionized calcium level and in calcium transients during in vitro development of normal and Duchenne muscular dystrophy cultured skeletal muscle measured by laser cytofluorimetry using indo-1. AB - Intracellular calcium activity was recorded during in vitro myogenesis of human normal and DMD muscle, using the calcium probe Indo-1 under laser illumination, at rest and during different kinds of stimulation (acetylcholine, high K+, caffeine). In myoblasts, the resting intracellular calcium level was significantly larger in DMD cells (89 +/- 9 nM; n = 40 vs 37 +/- 5 nM; n = 22) but there was no significant difference in myotubes, after fusion (44 +/- 4 nM; n = 34 vs 36 +/- 4 nM; n = 52). A similar evolution was observed in cells cultured from FSH biopsies. The amplitude of ACh- and high K(+)-induced calcium transients was significantly halved in DMD myotubes as compared to control ones and non significantly decreased for caffeine responses. Some alterations in the kinetics of responses were observed in DMD muscle: the rising phases of ACh- and high K(+) elicited transients and the decaying phase of the ACh-responses were significantly slowed down. It is concluded that: (i) in aneurally cultured human muscle, an increase in the basal level of internal calcium can occur at early stages of myogenesis before the expression of the dystrophin gene; and (ii) the changes in calcium transients induced by depolarization or direct stimulation of sarcoplasmic reticulum are not susceptible of inducing a calcium overload in DMD cells. PMID- 8402839 TI - Effect of percutaneous balloon mitral valvuloplasty on serum creatinine phosphokinase MB-isoenzyme levels. AB - Twenty-one patients with moderate to severe mitral stenosis were treated with percutaneous balloon mitral valvuloplasty (PBMV) using Inoue mitral double-lumen balloon catheters. Creatinine phosphokinase MB isoenzyme (CPK-MB) levels were measured at baseline, 5 min, 6 h, and 18 h post dilatation. Haemodynamic indexes, 12-lead ECGs, and 2-dimensional echocardiograms were performed to evaluate systolic function postprocedure. CPK-MB levels were modestly increased at 5 min and at 6 h postprocedure compared with the baseline level (p < 0.001) but were still within the normal reference range, except for one patient who had a level at 5 min minimally above the upper limit of normal. CPK-MB levels were not significantly increased at 18 h after the procedure. No significant changes in ejection fractions, 12-lead ECGs, and regional wall motion occurred in any of the patients studied. In conclusion, PBMV causes modest early elevation of CPK-MB. This elevation is not associated with changes in LV systolic function and does not interfere with the diagnosis of acute myocardial infarction using the enzyme elevation criteria. PMID- 8402840 TI - Percutaneous balloon mitral valvuloplasty: comparison of double and single (Inoue) balloon techniques. AB - To assess relative merits of the double and single (Inoue) balloon techniques of percutaneous balloon mitral valvuloplasty for treating symptomatic mitral stenosis, 33 patients who underwent dilatation with the double balloon were compared retrospectively with 28 who underwent dilatation with the Inoue balloon. There were no baseline differences in mean left atrial pressure, mean mitral gradient, or calculated mitral valve area between the two groups. The procedure was successful in 89% of patients in the double balloon group and in 93% in the Inoue balloon group. Final mitral valve area and absolute increase in mitral valve area were significantly greater in the double balloon group, in which there was a nonsignificant trend toward greater incidence of acute complications. Final left atrial pressure and mitral gradient were not different. Both techniques are useful and safe. PMID- 8402841 TI - Stenting of a renal artery stenosis achieves better relief of the obstructive lesion than balloon angioplasty. AB - A balloon-expandable (Palmaz-Schatz) stainless steel stent was utilized following balloon angioplasty (PTRA) to determine if the obstructive lesion, using quantitative methods (automated measuring the diameter stenosis, and transstenotic peak systolic and mean pressure gradients), was significantly further reduced or abolished. Hemodynamic transstenotic gradient and stenoses measurements were made during 21 renal artery stenting procedures; prior and following PTRA, and subsequent to stent deployment. The stent sizes placed in the renal arteries were 5 mm (19%), 6 mm (67%), and 7 mm (14%). The results were as follows: [table: see text] The balloon-expandable (Palmaz-Schatz) stent significantly further reduced, and in fact effectively abolished, the obstructive renal artery lesion in comparison to balloon angioplasty (PTRA). The stent's effectiveness with regard lesion recurrence, maintenance, and preservation of renal function; cure or improved management of hypertension; and survival will be determined by careful clinic follow-up. PMID- 8402842 TI - Coronary angioplasty with gradual versus rapid balloon inflation: initial results and complications. AB - Although a variety of coronary angioplasty balloon inflation protocols are employed, prior studies have not evaluated the relation of rate of inflation to the type and extent of arterial damage produced by angioplasty. We randomized 103 patients to either a gradual (gradual, incremental increase to peak inflation pressure) or rapid inflation protocol (rapid increase to peak inflation pressure). Fifty-one patients with 72 lesions underwent gradual and 52 patients with 73 lesions received rapid inflation protocols. There were no significant group differences with regard to age, sex, artery dilated, number of diseased vessels, presence of unstable angina and lesion morphological characteristics except for more lesions located on a bend in the gradual inflation group (p < 0.02). Although there was a tendency towards a higher success rate in patients with gradual inflation, the complete success rates were high in both groups (100% vs. 93%, p < 0.08). The dissection rate was higher in patients with rapid inflation (43/73 [59%] vs. 26/72 [36%], p < 0.01). The collective complication rate was higher in patients with rapid inflation (19% vs. 6%, p < 0.03). No deaths occurred in either group. Thus a gradual compared to rapid coronary angioplasty balloon inflation protocol reduces the frequency of dissection despite similar inflation pressure and balloon/vessel diameter ratio. Gradual inflations may reduce the frequency of procedure-related complications. PMID- 8402843 TI - Resistance of the atherosclerotic plaque during coronary angioplasty: a multivariate analysis of clinical and angiographic variables. AB - The atherosclerotic lesion resistance to balloon inflation was assessed prospectively in 200 patients undergoing primary single-site balloon percutaneous transluminal coronary angioplasty (PTCA) by using an identical inflation protocol. This resistance was evaluated by the stenosis resolution pressure, which is the pressure at which the lesion mark is no longer visible on the inflated balloon. The stenosis resolution pressure distribution was normal with a mean value of 4.4 +/- 2.3 atmospheres. Multivariate analysis revealed 4 factors related to the stenosis resolution pressure: stable angina pectoris (p < 10(-6)), the presence of calcifications (p = 0.016), the occurrence of vessel wall dissection after balloon angioplasty (p = 0.005), and the absence of a branch arising in the middle of the stenosis (p = 0.047). It is concluded that the mechanical resistance of the coronary lesion is related to the anginal status, the presence of calcifications, and the occurrence of vessel wall dissection after balloon angioplasty, probably on the basis of the plaque composition. PMID- 8402844 TI - Percutaneous left atrial to femoral arterial bypass pumping for circulatory support in high-risk coronary angioplasty. AB - Left atrial to femoral arterial bypass was evaluated as a means of supporting patients who were considered to be at high risk for the performance of percutaneous transluminal coronary angioplasty. A 20 French drainage catheter was inserted percutaneously into the left atrium via a modified transseptal technique. Blood was withdrawn from the left atrium and returned through a femoral arterial cannula using a roller pump. Thirteen patients were treated in this fashion with excellent circulatory support. Pump flows varied from 1.5 to 3 liters per minute and bypass time ranged from 27 to 106 min (mean = 43 +/- 17). Aortic mean pressure was well supported during balloon inflation. No significant complications were encountered. Neither an oxygenator nor a perfusionist is required. The ability to obtain direct left ventricular decompression offers a major potential advantage. Further evaluation of this technique for the support of such patients is indicated. PMID- 8402845 TI - Coronary angiography in cardiac myxomas: findings in 19 consecutive cases and review of the literature. AB - We reviewed the coronary angiographic findings of 19 patients with a cardiac myxoma, who underwent cardiac catheterization before surgery. Seventeen myxomas were localized in the left atrium and seven had angiographically visible tumor vascularity emerging from atrial branches of the right coronary artery in four patients and the circumflex coronary artery in three. In one patient, we found significant coronary artery disease of the circumflex coronary artery and in another we saw a thrombus-like lesion in the proximal third of the left anterior descending coronary artery. Our results are compared with the findings in two smaller groups of patients with cardiac myxoma who underwent coronary angiography preoperatively. We conclude that the major importance of coronary angiography in patients with cardiac myxomas is to exclude concomitant coronary artery disease before surgery. In a very small minority of patients, a selective coronary angiography is the clue to the diagnosis of cardiac myxoma. PMID- 8402846 TI - Rapid development of left main disease after coronary artery bypass grafting for failed angioplasty: a rare cause of postoperative angina. AB - We report a case of failed coronary angioplasty requiring urgent coronary artery bypass graft surgery. The return of angina pectoris early after surgery led to repeat catheterization, demonstrating a rapidly progressive stenosis of the left main coronary artery. This was treated successfully with coronary angioplasty. In patients with recurrent angina early after bypass surgery in whom angioplasty preceded surgery, acceleration of left main disease should be considered. PMID- 8402847 TI - Inflated balloon entrapped in a calcified coronary stenosis. AB - Balloon angioplasty in calcified coronary lesions may have a decreased success rate and an increased incidence of complications. Previous cases have been reported on the phenomena of catheter or wire entrapment, but this report highlights a new problem in association with a calcified stenoses. This case lead to the unusual problem of the inflated balloon being entrapped within the lesion while the distal portion of the balloon remained inflated even after balloon deflation. PMID- 8402848 TI - Collateralized chronic total occlusion: a different animal. AB - This report describes a patient with a chronically occluded proximal LAD in whom antegrade flow was not re-established by successful dilatation alone. Obliteration of retrograde collateral flow restored antegrade flow. This case demonstrates the potential detrimental effect of vigorous collateral flow on the patency of chronic occlusions after successful PTCA. PMID- 8402849 TI - Coronary angioplasty of anomalous right coronary arteries. AB - We report 2 cases of successful angioplasty of anomalous right coronary arteries originating above the sinotubular line at the junction of the right and left sinus of Valsalva. The use of Amplatz left guiding catheters provided optimal support for performing angioplasty. PMID- 8402850 TI - Anomalous connection between the sinus node artery and the A-V node artery. AB - Anomalous connection between the sinus node artery and A-V node artery is an extremely rare coronary variant. Angiographic and clinical data from an adult with this finding are reported. Coronary embryogenesis and normal nodal arterial blood supply are reviewed. PMID- 8402851 TI - Can an intracoronary Doppler wire accurately measure changes in coronary blood flow velocity? AB - The purpose of this study was to compare a newly developed Doppler guide wire in the measurement of coronary blood flow velocity (CBFV) and coronary flow reserve (CFR) when compared to a previously extensively validated Doppler catheter. Sequential measurements of Doppler signals from the Doppler catheter and wire were obtained in response to graded doses of intracoronary adenosine in the left circumflex of 10 anesthetized dogs. The Doppler catheter signal was recorded on a standard physiologic chart recorder; that from the Doppler wire was recorded both on the chart recorder and the commercially available spectral analysis recording system. High correlations were obtained between the change in coronary velocity and CFR which were independent of the recording system used. Peak flow reserve in response to maximum intracoronary adenosine (24 micrograms bolus) ranged from 3.7 5.6 x basal. These results are similar to those obtained in normal humans. In spite of major differences between engineering specifications of the Doppler wire and Doppler catheter, under optimal conditions these two techniques yield similar results. The Doppler wire may facilitate measurements of CFR during diagnostic angiography in small or diffusely diseased vessels. PMID- 8402852 TI - Coronary angioplasty using an autoperfusion balloon catheter through a 6 French guiding catheter. AB - Use of 6 French guiding catheters for elective percutaneous transluminal coronary angioplasty has been limited by lack of a compatible autoperfusion balloon catheter for management of complications such as acute vessel closure and large subintimal dissections. We describe the successful use of a lower profile autoperfusion balloon catheter through large internal lumen 6F guiding catheters for elective coronary angioplasty. These cases demonstrate the feasibility of the use of autoperfusion balloon catheters with 6F guiding catheters in elective, and presumably also in emergent, settings. PMID- 8402853 TI - New techniques for guiding catheter management during directional coronary atherectomy. AB - Directional coronary atherectomy (DCA) is an important advance in the mechanical revascularization of stenotic coronary arteries. The bulky nature of the DCA device has necessitated the use of guiding catheter designs that are more cumbersome to use than balloon angioplasty guiding catheters. Because engagement of coronary artery ostia with the currently available DCA guiding catheters is often difficult and because DCA guiding catheters significantly "relax" and reshape during the atherectomy procedure, angiography using these guiding catheters before and after atherectomy can be suboptimal. A new technique for angiography during atherectomy using long Judkin's diagnostic catheters inserted through the existing DCA guiding catheters is described. This technique can be used for optimal visualization of the coronary arteries with minimal use of contrast before and after sessions of atherectomy and also can be used to help engage the DCA guiding catheters. Some improvements in the design of guiding catheters for DCA are suggested. PMID- 8402854 TI - Left ventricular rupture following myocardial infarction treated with streptokinase: successful resuscitation in the cardiac catheterization laboratory using pericardiocentesis and autotransfusion. AB - An 80-year-old female developed left ventricular rupture in the cardiac catheterization laboratory following myocardial infarction treated with streptokinase. The patient was resuscitated using pericardiocentesis and autotransfusion. This technique and the predisposing factors for left ventricular rupture are discussed. PMID- 8402855 TI - Guidelines for training, credentialing, and maintenance of competence for the performance of coronary angioplasty: a report from the Interventional Cardiology Committee and the Training Program Standards Committee of the Society for Cardiac Angiography and Interventions. PMID- 8402856 TI - Radiation exposure: comparison of rapid exchange and conventional over-the-wire coronary angioplasty systems. AB - The growth of coronary angioplasty has resulted in increased fluoroscopy time to patients, staff, and physicians. Rapid exchange-type catheters have purported to reduce fluoroscopy time and procedure time compared to conventional over-the-wire systems. Of 150 consecutive patients, 54 were treated solely with rapid-exchange catheters and 84 were treated solely with over-the-wire catheters. Excluding 12 cases treated with fixed wire or combination catheters, the following data were found: [table: see text] Overall procedural success was obtained in 98.5% (136/138) of patients, 1.5% (2) requiring emergency coronary artery bypass graft surgery. A significant 35% reduction in fluoroscopy time and 13% reduction in procedure time were found when rapid-exchange catheters were used, with identical success rates, number of balloons used per case, and complication rates compared to over-the-wire catheters. Rapid-exchange catheters should be considered as part of an overall effort to reduce radiation exposure in the interventional cardiology laboratory. PMID- 8402857 TI - Simplified method for calculating aortic valve resistance: correlation with valve area and standard formula. AB - Aortic valve resistance (AVR) is a useful index to assess the severity of aortic stenosis. This study compared the standard method to calculate AVR with a simplified method based on the conventional approach for measuring vascular resistance: AVR = (peak-to-peak transaortic pressure gradient/(cardiac output*2.5))*80, where 80 is a conversion factor and 2.5 assumes that the systolic ejection period comprises 40% of the R-R cycle. We compared the standard AVR, the simplified AVR, and the Gorlin-derived value area in 118 patients with pure or dominant aortic stenosis. There was a strong linear correlation between the standard and simplified AVR (r = 0.96, p < .0001). There was a curvilinear relation between the aortic valve area and AVR (r = 0.92, p < .001). In 48 patients with aortic valve area > or = 0.7 cm2, the AVR was < 300 dynes-sec-cm-5 in 45 patients (94%) by the standard method and in 42 patients (88%) by the simplified method (p = NS). In conclusion, our method for measuring AVR is accurate and simpler than the standard method. PMID- 8402858 TI - Coronary angioplasty through 4 French diagnostic catheters. AB - In 50 consecutive patients subjected to coronary angioplasty immediately following a 4 French (F) diagnostic study, the technical feasibility and economical aspects of angioplasty through 4F catheters of 54 lesions were assessed. The patients were selected, but multiple, eccentric, and long lesions were not a priori excluded. 4F diagnostic catheters (Cordis), and fixed-wire dilatation catheters (Ace, Scimed) were used in all cases. The procedure was successful in 43 lesions (80%) using 4F catheters. For 11 stenoses (20%), a change over to a larger French size was required. Two of these lesions could not be crossed with the balloon despite the larger sized guiding catheter. The final overall success rate was 96%, and there were no major complications. The use of diagnostic 4F catheters for angioplasty in these 50 patients resulted in the saving of 39 guiding catheters and 19 introducer sheaths. For 12 lesions (22%), an additional 4F catheter became necessary since the shape used for the diagnostic study was inadequate for angioplasty. In 7 cases, more than 1 balloon was used, but 5 of these balloon exchanges were independent of the use of 4F catheters. Three exchanges were performed through the 4F catheter (1 for need of a larger balloon to improve on an unsatisfactory angiographic result and 2 for a crimped guide wire tip of the Ace balloon). In the remaining 4, a larger catheter was used; in 2 of them, angioplasty eventually failed (failure to cross lesion) and in the remaining 2, a Monorail system solved the problem, which is incompatible with 4F catheters.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8402859 TI - Fragmentation and embolization of a plastic-coated guidewire. AB - We report a case of fragmentation and embolization to the pulmonary artery of sheared pieces of a recently introduced hydrophilic plastic-coated guidewire. The propensity for this to occur, the lack of initial recognition, and the ability to retrieve the fragments by catheter techniques are emphasized. As these guidewires are more frequently used, physicians should be aware of this potential complication. PMID- 8402860 TI - Catheter atherectomy of intimal fibroplasia of the common iliac artery. AB - An 18-year-old woman presented with renovascular hypertension and left lower extremity claudication. Aorto-iliac angiography showed stenotic lesions in the left renal artery and the left common iliac artery. For uncontrolled hypertension, nephrectomy was performed and histopathology of the renal artery showed intimal fibroplasia, an uncommon type of fibromuscular dysplasia. The left common iliac artery lesions were treated with directional atherectomy, which produced excellent immediate angiographic and symptomatic improvement. PMID- 8402861 TI - Bail-out coronary stenting in an extremely tortuous right coronary artery with the Palmaz-Schatz stent and Teleguide sheath. AB - Significant vessel tortuosity is a relative contraindication to the use of the Palmaz-Schatz coronary stent for fear of stent displacement during delivery. We describe a patient with unstable angina in whom conventional coronary angioplasty in an extremely tortuous right coronary artery resulted in an occlusive dissection. Emergency bypass surgery was avoided by the successful placement of a protected Palmaz-Schatz stent using a 5F Teleguide sheath. PMID- 8402862 TI - Percutaneous transluminal coronary angioplasty of a right coronary artery arising from the left main coronary artery. AB - Angioplasty of anomalous coronary arteries presents unique technical challenges. Correct guiding catheter selection is important to ensure adequate access to the anomalous vessel and to provide support to cross the lesion. A case of successful PTCA of a lesion in an anomalous right coronary artery arising from the left main coronary artery is presented. PMID- 8402863 TI - Percutaneous transvenous mitral commissurotomy in patients with severe kyphoscoliosis. AB - Because transseptal catheterization is felt to be contraindicated in patients with severe kyphoscoliosis, there have been no reports of percutaneous transvenous mitral commissurotomy performed in such patients. This report describes percutaneous transvenous mitral commissurotomy in three patients with severe thoracic kyphoscoliosis, with special emphasis on the transseptal puncture technique. Biplane right atrial angiography and the contrast septal flush method are very useful in landmark selection for a safe transseptal puncture. PMID- 8402864 TI - Double left anterior descending artery originating from the left and right coronary artery: a rare coronary artery anomaly. AB - A case of double left anterior descending coronary artery is presented. The double artery originated from the left main stem and the right coronary artery. There were no stenoses on these two arteries. This anomaly of coronary arteries seems to be very rare. PMID- 8402865 TI - Pseudocoarctation of the aorta: a magnetic resonance imaging correlation. AB - We present here the clinical and angiographic features in a patient with pseudocoarctation of the aorta and aortic valvular stenosis. An MRI study performed to delineate the aorta provided valuable correlations and highlighted the potential of this noninvasive modality for the diagnosis of pseudocoarctation of the aorta. PMID- 8402866 TI - Application of intracoronary flow velocity for detection and management of ostial saphenous vein graft lesions. AB - Ostial lesions of saphenous vein grafts can be difficult to assess by angiography. A physiologic approach to the selection and recanalization of ostial lesions in saphenous vein grafts may be advantageous to overcome limitations of angiography. To assist in identifying favorable physiology and facilitate procedural decision making, the use of coronary flow velocity measurements with a Doppler-tipped 0.018-inch angioplasty flowire in three particularly difficult patients with varying types of saphenous vein graft aorto-ostial narrowings is reported. PMID- 8402867 TI - Evaluation of two oximeters for use in cardiac catheterization laboratories. AB - We evaluated two whole-blood oximeters designed for use in the cardiac catheterization laboratory: the Oxicom 3000 and the AVOXimeter 1000. Unlike the larger CO-Oximeters, which hemolyze a blood sample before analysis, these simple instruments use disposable cuvettes to allow determination of oxyhemoglobin saturation of whole blood. Thus they eliminate the need for cleaning solutions and considerable maintenance. We evaluated the accuracy of these instruments in comparison with a Radiometer OSM3 and found them both to work well on routine samples. Of note, the AVOXimeter performed better than the Oxicom when the sample was hemolyzed and when green dye was present. In addition, the AVOXimeter measures the total hemoglobin concentration and the oxygen content of each sample, eliminating errors that hemodilution introduces into the determination of cardiac output by the Fick principle. We conclude that whole-blood oximeters are accurate and useful instruments for use in the cardiac catheterization laboratory. PMID- 8402868 TI - In vitro evaluation of blood flow through autoperfusion balloon catheters. AB - The effective flow rates with human blood through an autoperfusion catheter cannot be monitored in vivo and have not been experimentally determined in vitro. The manufacturers (Advanced Cardiovascular Systems [ACS], Temecula, CA) have suggested that "the flow rate" through the Stack over the wire and the RX-60 monorail catheter is 60 ml/min with a pressure gradient of 80 mmHg. We measured human blood flow rates in vitro through these catheters under different continuous pressure regimens (between 40 and 120 mmHg), with varying hematocrit levels (between 25% and 62%). Measured blood flows at a gradient of 80 mmHg were found to vary from 32 to 65 cc/min, with hematocrit levels of 62-25%. Minor variations in the circuitry, besides the viscosity of the medium, cause significant changes in observed flow rates (such as kinking of the catheter and blood sedimentation). In vitro determinations of blood flows cannot automatically be transferred to the in vivo condition, primarily because in vitro determinations do not account for the systolic intramural pressure increase (which may overcome the aortic pressure). If such a phenomenon is also considered, then the in vitro flow rates reported here should be multiplied by a factor of 0.40-0.60 to determine effective in vivo flow rates. Such information is relevant for the clinical operator of angioplasty, especially in the treatment of patients at high risk for undergoing percutaneous transluminal coronary angioplasty. PMID- 8402869 TI - Retrieval techniques for managing flexible intracoronary stent misplacement. AB - With the increasing use of flexible intracoronary stents, the likelihood of complications, including stent misplacement, will tend to rise. We describe the successful use of three commercially available retrieval devices: the nitinol gooseneck snare, the biliary forceps, and the multipurpose basket. We recommend the availability of these devices to operators involved in intracoronary stent placement. PMID- 8402870 TI - Brachial approach directional coronary atherectomy of a left coronary artery saphenous vein bypass graft. AB - Directional coronary atherectomy (DCA) of a saphenous vein bypass graft to the left coronary artery was performed percutaneously from the brachial artery approach using a 7F endomyocardial biopsy sheath. Initial positioning was accomplished with a left bypass graft catheter inserted in the sheath. This technique permits use of smaller catheters than usual for DCA in patients in whom larger guides cannot be used. PMID- 8402871 TI - Facilitated drainage of pericardial effusion with a fenestrated pigtail catheter and sheath system. PMID- 8402872 TI - Transcatheter management of pulmonary venous pathway obstruction with atrial baffle leak following Mustard and Senning repair. AB - Two patients presenting with pulmonary venous baffle obstruction following Mustard or Senning repair of transposition of the great arteries were successfully treated with percutaneous balloon dilatation. At the time of baffle dilatation, a significant systemic to pulmonary venous Mustard baffle leak was successfully closed with a Rashkind ductal occluder device. Specific features pertaining to the morphology of the baffle defect that allowed successful catheter occlusion are discussed and compared to that found in a Senning patient. The important adjunctive role of transoesophageal echocardiography within the catheterization laboratory is emphasized. PMID- 8402873 TI - Experience with a double loop guiding catheter for angioplasty of the right coronary artery. AB - An 8 French (F) double loop guiding (DLG) catheter was developed for percutaneous transluminal coronary angioplasty (PTCA) of the right coronary artery (RCA) and tested in 80 patients: primary use in 59 patients, and after failure of an 8 F right Judkins guiding (RJG) catheter in 21 patients. Primary use resulted in stable intubation of RCA in 55 patients (93%), but PTCA was successful in 50 (91%). Five failures resulted from inability to cross or dilate the lesion. After failure of an 8 F RJG in 21 patients, successful stable intubation of RCA with a DLG was achieved in 19 patients (90%), but PTCA was successful in 17 patients (81%). Failure of RJGs in 21 patients resulted from inability to intubate the RCA in 12 patients, or inadequate back up support by the guiding in 9 patients. CONCLUSION: DLGs increased the success rate of PTCA of the RCA after failure of RJGs. PMID- 8402874 TI - Recanalization in femoral arterial occlusions. PMID- 8402875 TI - Acute aortic regurgitation. PMID- 8402876 TI - Use of 3-way stopcock as a torquer for PTCA. PMID- 8402877 TI - Miles to go before I sleep. PMID- 8402878 TI - TGF beta inhibition of Cdk4 synthesis is linked to cell cycle arrest. AB - Transforming growth factor beta 1 (TGF beta 1) causes G1 growth arrest and the accumulation of unphosphorylated retinoblastoma protein (Rb) in responsive cells. Cdk4 (cyclin-dependent kinase), a major catalytic subunit of the mammalian D-type G1 cyclins, can phosphorylate Rb in vitro, and at least one D-type cyclin, D2, directs the phosphorylation of Rb in vivo. Here we show that TGF beta 1 induces suppression of cdk4 synthesis in G1 in mink lung epithelial cells. Constitutive cdk4 synthesis in these cells led to TGF beta 1 resistance. It also resulted in growth in low serum medium when these cells were released from contact inhibition. Cdk2 activity was also suppressed by TGF beta 1 action, but its constitutive expression failed to override a TGF beta 1-induced G1 block. Hence, the TGF beta 1 block is primarily mediated by cdk4 modulation. Further evidence suggests that TGF beta 1-induced down-modulation of cdk4 leads to inhibition of cdk2 activation and that both events might contribute to TGF beta 1 growth suppression. PMID- 8402879 TI - Resolution of Holliday junctions by RuvC resolvase: cleavage specificity and DNA distortion. AB - E. coli RuvC protein resolves Holliday junctions during genetic recombination and postreplication repair. Using small synthetic junctions, we show that junction recognition is structure-specific and occurs in the absence of metal cofactors. In the presence of Mg2+, Holliday junctions are resolved by the introduction of symmetrically related nicks at the 3' side of thymine residues. The nicked duplex products are repaired by the action of DNA ligase. Within the RuvC-Holliday junction complex, the DNA is distorted such that 2 of the 4 strands become hypersensitive to hydroxyl radical attack. The ionic requirements of binding, hydroxyl radical sensitivity, and strand cleavage indicate three distinct steps in the mechanism of RuvC-mediated Holliday junction resolution: structure specific recognition, DNA distortion, and sequence-dependent cleavage. PMID- 8402880 TI - A receptor for subgroup A Rous sarcoma virus is related to the low density lipoprotein receptor. AB - Cellular receptors are required for efficient entry of retroviruses into cells. We previously cloned a chicken gene responsible for susceptibility to the retrovirus subgroup A Rous sarcoma virus (RSV(A)). Here we have isolated the quail homolog and generated two alternatively spliced processed genes encoding cellular receptors for RSV(A). Predicted products of the processed genes appear to be small membrane-associated proteins with identical 83 amino acid extracellular domains but different membrane anchors. Within the extracellular domain is a region closely related to the ligand-binding repeat of the low density lipoprotein receptor (LDLR). Expression of either processed gene renders mammalian cells specifically susceptible to RSV(A). Antibodies directed against the receptor block subgroup A infection of avian cells via endogenous receptors and have no effect on entry of other RSV subgroups. Thus, small LDLR-related proteins are cellular receptors for RSV(A). PMID- 8402881 TI - Common signals control low density lipoprotein receptor sorting in endosomes and the Golgi complex of MDCK cells. AB - The cytoplasmic domain of the LDL receptor bears two tyrosine-containing determinants that can independently target receptors from the Golgi to the basolateral plasma membrane of MDCK cells. We found that these determinants, localized to the membrane-proximal and -distal regions of the receptor's cytoplasmic domain, also control polarized sorting in endosomes. Inactivation of the distal determinant reduced receptors' ability to return to the basolateral domain following endocytosis, resulting instead in receptor transcytosis from basolateral endosomes to the apical plasma membrane. Similarly, receptors internalized from the apical surface were transported from apical endosomes to the basolateral surface, owing to the proximal basolateral targeting determinant. Thus, receptor recycling in endosomes is directed by the same signals as polarized sorting in the Golgi, indicating that sorting on the endocytic and biosynthetic pathways involves similar mechanisms. The observation that brefeldin A interfered with sorting but not transport in both endosomes and the Golgi further supports this. PMID- 8402882 TI - Following a diabetogenic T cell from genesis through pathogenesis. AB - Nonobese diabetic (NOD) mice spontaneously develop a disease very similar to type 1 diabetes in humans. We have generated a transgenic mouse strain carrying the rearranged T cell receptor genes from a diabetogenic T cell clone derived from a NOD mouse. Self-reactive T cells expressing the transgene-encoded specificity are not tolerized in these animals, resulting in rampant insulitis and eventually diabetes. Features of the disease process emphasize two so-called check-points, recognized previously in the NOD and human diseases but easily misinterpreted. Although NOD mice are protected from insulitis and diabetes by expression of the E molecule encoded in the major histocompatibility complex, the transgenics are not, permitting us to exclude some possible mechanisms of protection. PMID- 8402883 TI - Transcription factor p91 interacts with the epidermal growth factor receptor and mediates activation of the c-fos gene promoter. AB - Transcription factor p91 contains a SH2 domain and is activated by tyrosine phosphorylation. Here we demonstrate that epidermal growth factor (EGF) induces rapid tyrosine phosphorylation and nuclear translocation of p91. Through its SH2 domain, p91 directly interacts with the EGF receptor in a ligand-dependent manner. p91 is a necessary component of an EGF-induced DNA-binding factor that recognizes a previously identified regulatory element, SIE (c-sis-inducible element), in the c-fos gene promoter. Activated p91 stimulates SIE-dependent transcription in vitro. Cotransfection of an SIE-containing reporter with a p91 expression vector shows that p91 is a positive transcriptional regulator of the c fos gene promoter. These studies suggest that EGF uses a direct signaling pathway to control nuclear transcriptional events. PMID- 8402884 TI - Lipo-oligosaccharide nodulation factors: a minireview new class of signaling molecules mediating recognition and morphogenesis. PMID- 8402885 TI - p53-dependent apoptosis modulates the cytotoxicity of anticancer agents. AB - Although the primary cellular targets of many anticancer agents have been identified, less is known about the processes leading to the selective cell death of cancer cells or the molecular basis of drug resistance. p53-deficient mouse embryonic fibroblasts were used to examine systematically the requirement for p53 in cellular sensitivity and resistance to a diverse group of anticancer agents. These results demonstrate that an oncogene, specifically the adenovirus E1A gene, can sensitize fibroblasts to apoptosis induced by ionizing radiation, 5 fluorouracil, etoposide, and adriamycin. Furthermore, the p53 tumor suppressor is required for efficient execution of the death program. These data reinforce the notion that the cytotoxic action of many anticancer agents involves processes subsequent to the interaction between drug and cellular target and indicate that divergent stimuli can activate a common cell death program. Consequently, the involvement of p53 in the apoptotic response suggests a mechanism whereby tumor cells can acquire cross-resistance to anticancer agents. PMID- 8402886 TI - Small molecules that selectively block RNA binding of HIV-1 Rev protein inhibit Rev function and viral production. AB - Replication of RNA viruses, such as the human immunodeficiency virus (HIV), is dependent upon multiple specific interactions between viral RNAs and viral and cellular proteins. A small molecule that interferes specifically with one or more of these RNA-protein interactions could be an efficacious antiviral agent. Here we show that certain aminoglycoside antibiotics, in particular neomycin B, can block binding of the HIV Rev protein to its viral RNA recognition element. Inhibition appears to be highly selective, resulting from competitive binding of the drug to a small viral RNA region within the Rev-binding site. We further demonstrate that neomycin B can specifically antagonize Rev function in vitro and in vivo and can inhibit production of HIV. Our results establish the feasibility for developing antiviral drugs that act by selectively blocking RNA-protein interactions. PMID- 8402887 TI - Reversal of terminal differentiation and control of DNA replication: cyclin A and Cdk2 specifically localize at subnuclear sites of DNA replication. AB - DNA replication in mammalian cells occurs in discrete nuclear foci. Here we show that terminally differentiated myotubes can be induced to reenter S phase and show the same pattern of replication foci as cycling cells. We used this cellular system to analyze the interaction of cell cycle proteins with these foci in vivo. Cyclin A and cdk2, but not cyclin B1 and cdc2, were specifically localized at nuclear replication foci, just like the replication protein proliferating cell nuclear antigen. A potential target of cyclin A and cdk2 is the 34 kd subunit of replication protein A (RPA34). In contrast with the 70 kd subunit, which localizes to the foci, RPA34 was not detected at these replication sites, which may reflect a transient interaction. The specific localization of cyclin A and cdk2 at nuclear replication foci provides a direct link between cell cycle regulation and DNA replication. PMID- 8402888 TI - Mechanisms that help the yeast cell cycle clock tick: G2 cyclins transcriptionally activate G2 cyclins and repress G1 cyclins. AB - In budding yeast, G1 cyclins such as CLN1 and CLN2 are expressed in G1 and S phases, while mitotic cyclins such as CLB1 and CLB2 are expressed in G2 and M phases. We find that the CLBs play a central role in the transition from CLNs to CLBs: the CLBs stimulate their own expression while repressing that of CLNs. This negative regulation of CLNs may occur via the transcription factor SWI4, because CLBs are necessary for G2 repression of SCB-regulated genes like CLN1 and CLN2 but not for repression of MCB-regulated genes like DNA polymerase and CLB5. Furthermore, SW14 associates with CLB2 protein and is a substrate for the CLB2 associated CDC28 kinase in vitro. PMID- 8402889 TI - Membrane fusion machinery: insights from synaptic proteins. PMID- 8402890 TI - Targeted disruption of the trkB neurotrophin receptor gene results in nervous system lesions and neonatal death. AB - We have generated mice carrying a germline mutation in the tyrosine kinase catalytic domain of the trkB gene. This mutation eliminates expression of gp145trkB, a protein-tyrosine kinase that serves as the signaling receptor for two members of the nerve growth factor family of neurotrophins, brain-derived neurotrophic factor and neurotrophin-4. Mice homozygous for this mutation, trkBTK(-/-), develop to birth. However, these animals do not display feeding activity, and most die by P1. Neuroanatomical examination of trkBTK (-/-) mice revealed neuronal deficiencies in the central (facial motor nucleus and spinal cord) and peripheral (trigeminal and dorsal root ganglia) nervous systems. These findings illustrate the role of the gp145trkB protein-tyrosine kinase receptor in the ontogeny of the mammalian nervous system. PMID- 8402891 TI - Golgi membrane dynamics imaged by freeze-etch electron microscopy: views of different membrane coatings involved in tubulation versus vesiculation. AB - We used high resolution three-dimensional electron microscopy to visualize Golgi cisternal structure and analyze morphological transitions induced by various in vitro incubations. Our images show that Golgi cisternae have two distinct surface coatings with different distributions and apparent functions. The first type, probably a coatomer coat, consists of tightly packed approximately 10 nm surface particles. These are localized exclusively to sites of membrane budding and are as intimately involved in bud formation as clathrin or caveolin coats. When this coating is exaggerated by GTP gamma S, the periphery of all cisternae is partitioned into coated vesicles that remain attached at their sites of formation. A second, much finer coating is evenly distributed over the periphery of cisternae, including tubules enclosing the fenestrae. It appears to stabilize the membrane curvature associated with tubules and edges. These different coatings must be considered in further attempts to unravel Golgi membrane trafficking mechanisms. PMID- 8402892 TI - Brain myosin-V is a two-headed unconventional myosin with motor activity. AB - Chicken myosin-V is a member of a recently recognized class of myosins distinct from both the myosins-I and the myosins-II. We report here the purification, electron microscopic visualization, and motor properties of a protein of this class. Myosin-V molecules consist of two heads attached to an approximately 30 nm stalk that ends in a globular region of unknown function. Myosin-V binds to and decorates F-actin, has actin-activated magnesium-ATPase activity, and is a barbed end-directed motor capable of moving actin filaments at rates of up to 400 nm/s. Myosin-V does not form filaments. Each myosin-V heavy chain is associated with approximately four calmodulin light chains as well as two less abundant proteins of 23 and 17 kd. PMID- 8402893 TI - Complementation cloning of an MHC class II transactivator mutated in hereditary MHC class II deficiency (or bare lymphocyte syndrome). AB - Hereditary major histocompatibility complex (MHC) class II deficiency (or bare lymphocyte syndrome) is a form of severe primary immunodeficiency with a total lack of MHC class II expression. It is due to a defect in the regulation of MHC class II genes. A novel gene was isolated by complementation cloning, using an MHC class II-negative mutant cell line. This gene (CIITA) functions as a transactivator of MHC class II gene expression and restores expression of all MHC class II isotypes in mutant cells. In addition, CIITA fully corrects the MHC class II regulatory defect of cells from patients with bare lymphocyte syndrome. In this disease we have identified a splicing mutation that results in a 24 amino acid deletion in CIITA, resulting in loss of function of the transactivator. Hence, the CIITA gene is essential for MHC class II gene expression and has been shown to be responsible for hereditary MHC class II deficiency. PMID- 8402894 TI - Bacteriophage T4 Alc protein: a transcription termination factor sensing local modification of DNA. AB - Bacteriophage T4 Alc protein participates in shutting off host transcription after infection of E. coli. It is demonstrated that Alc acts as a site-specific termination factor. The Alc sites occur frequently in E. coli DNA, resulting in early cessation of elongation in several tested transcription units. Alc dependent termination requires unimpeded propagation of the elongating complex as it approaches the Alc site. Temporary halting of RNA polymerase within 10-15 bp before the Alc site prevents termination. Bacteriophage T4 transcription is protected from the action of Alc by overall substitution of cytosine with 5 hydroxymethyl cytosine in T4 DNA. In vitro methylation of CpG sequences in the vicinity of an Alc site abolishes the effect of Alc. Thus, Alc-dependent termination involves local sensing of the state of cytosine modification and a short-term "memory" of recent pausing. PMID- 8402895 TI - Functional analysis of the P box, a domain in cyclin B required for the activation of Cdc25. AB - Cyclin B-cdc2 complexes are kept inactive by inhibitory phosphorylations on Thr 14 and Tyr-15 of cdc2 until they are dephosphorylated at the end of G2 by the phosphatase cdc25. Recent work has suggested that a small region of cyclin B, which we call the P box, may contribute part of a phosphatase-activating domain to cdc25. Individual conservative substitutions at three invariant residues within the P box yield mutant cyclin B proteins that bind cdc2 in vitro and then show the predicted cell cycle arrest, with cdc25 remaining in the low activity interphase form and cyclin B-cdc2 complexes remaining phosphorylated and inactive. While the low activity interphase form of cdc25 cannot act on cdc2 complexed with a mutant P box cyclin, the high activity M phase form of cdc25 can. These results demonstrate that the P box domain of cyclin B is required for cdc25 activation and support a two-step mechanism for the cdc25-dependent activation of cyclin B-cdc2. PMID- 8402896 TI - BCR-ABL-induced oncogenesis is mediated by direct interaction with the SH2 domain of the GRB-2 adaptor protein. AB - BCR-ABL is a chimeric oncoprotein that exhibits deregulated tyrosine kinase activity and is implicated in the pathogenesis of Philadelphia chromosome (Ph1) positive human leukemias. Sequences within the first exon of BCR are required to activate the transforming potential of BCR-ABL. The SH2/SH3 domain-containing GRB 2 protein links tyrosine kinases to Ras signaling. We demonstrate that BCR-ABL exists in a complex with GRB-2 in vivo. Binding of GRB-2 to BCR-ABL is mediated by the direct interaction of the GRB-2 SH2 domain with a phosphorylated tyrosine, Y177, within the BCR first exon. The BCR-ABL-GRB-2 interaction is required for activation of the Ras signaling pathway. Mutation of Y177 to phenylalanine (Y177F) abolishes GRB-2 binding and abrogates BCR-ABL-induced Ras activation. The BCR-ABL (Y177F) mutant is unable to transform primary bone marrow cultures and is impaired in its ability to transform Rat1 fibroblasts. These findings implicate activation of Ras function as an important component in BCR-ABL-mediated transformation and demonstrate that GRB-2 not only functions in normal development and mitogenesis but also plays a role in oncogenesis. PMID- 8402897 TI - SREBP-1, a basic-helix-loop-helix-leucine zipper protein that controls transcription of the low density lipoprotein receptor gene. AB - Sterol regulatory element 1 (SRE-1), a decamer (5'-ATC-ACCCCAC-3') flanking the low density lipoprotein (LDL) receptor gene, activates transcription in sterol depleted cells and is silenced by sterols. We report the cDNA cloning of human SREBP-1, a protein that binds SRE-1, activates transcription, and thereby mediates the final regulatory step in LDL metabolism. SREBP-1 contains a basic helix-loop-helix-leucine zipper (bHLH-ZIP) motif, but it differs from other bHLH ZIP proteins in its larger size (1147 amino acids) and target sequence. Instead of an inverted repeat (CANNTG), the target for all known bHLH-ZIP proteins, SRE-1 contains a direct repeat of CAC. Overexpression of SREBP-1 activates transcription of reporter genes containing SRE-1 in the absence (15-fold) and presence (90-fold) of sterols, abolishing sterol regulation. We suggest that SREBP-1 is regulated by an unknown factor that is overwhelmed when SREBP-1 is overexpressed. Understanding the regulation of SREBP-1 may be crucial for understanding the control of plasma cholesterol in humans. PMID- 8402898 TI - The GTPase dynamin binds to and is activated by a subset of SH3 domains. AB - Src homology 3 (SH3) domains have been implicated in mediating protein-protein interactions in receptor signaling processes; however, the precise role of this domain remains unclear. In this report, affinity purification techniques were used to identify the GTPase dynamin as an SH3 domain-binding protein. Selective binding to a subset of 15 different recombinant SH3 domains occurs through proline-rich sequence motifs similar to those that mediate the interaction of the SH3 domains of Grb2 and Abl proteins to the guanine nucleotide exchange protein, Sos, and to the 3BP1 protein, respectively. Dynamin GTPase activity is stimulated by several of the bound SH3 domains, suggesting that the function of the SH3 module is not restricted to protein-protein interactions but may also include the interactive regulation of GTP-binding proteins. PMID- 8402900 TI - Decoding the nucleosome. PMID- 8402899 TI - Identification of a residue in the translocation pathway of a membrane carrier. AB - Preliminary work using directed mutagenesis proved that cysteine is not required for operation of UhpT, the anion exchange protein responsible for glucose 6 phosphate transport by E. coli. We then made a detailed study of C143 and C265, because these cysteines impart sensitivity to p-chloromercuribenzosulfonate (PCMBS), a sulfhydral agent resembling glucose 6-phosphate in size, shape, and charge. We showed that C143 was exposed to the cytoplasm, as expected from hydropathy analysis, but we found no sidedness for C265. Rather, C265 was accessible to PCMBS from both membrane surfaces. And since the attack at C265 was blocked by glucose 6-phosphate, position 265 must lie directly on the pathway taken by the substrate as it moves through this membrane carrier. PMID- 8402901 TI - Mice carrying null mutations of the genes encoding insulin-like growth factor I (Igf-1) and type 1 IGF receptor (Igf1r). AB - Newborn mice homozygous for a targeted disruption of insulin-like growth factor gene (Igf-1) exhibit a growth deficiency similar in severity to that previously observed in viable Igf-2 null mutants (60% of normal birthweight). Depending on genetic background, some of the Igf-1(-/-) dwarfs die shortly after birth, while others survive and reach adulthood. In contrast, null mutants for the Igf1r gene die invariably at birth of respiratory failure and exhibit a more severe growth deficiency (45% normal size). In addition to generalized organ hypoplasia in Igf1r(-/-) embryos, including the muscles, and developmental delays in ossification, deviations from normalcy were observed in the central nervous system and epidermis. Igf-1(-/-)/Igf1r(-/-) double mutants did not differ in phenotype from Igf1r(-/-) single mutants, while in Igf-2(-)/Igf1r(-/-) and Igf-1( /-)/Igf-2(-) double mutants, which are phenotypically identical, the dwarfism was further exacerbated (30% normal size). The roles of the IGFs in mouse embryonic development, as revealed from the phenotypic differences between these mutants, are discussed. PMID- 8402902 TI - Role of insulin-like growth factors in embryonic and postnatal growth. AB - A developmental analysis of growth kinetics in mouse embryos carrying null mutations of the genes encoding insulin-like growth factor I (IGF-I), IGF-II, and the type 1 IGF receptor (IGF1R), alone or in combination, defined the onset of mutational effects leading to growth deficiency and indicated that between embryonic days 11.0 and 12.5, IGF1R serves only the in vivo mitogenic signaling of IGF-II. From E13.5 onward, IGF1R interacts with both IGF-I and IGF-II, while IGF-II recognizes an additional unknown receptor (XR). In contrast with the embryo proper, placental growth is served exclusively by an IGF-II-XR interaction. Additional genetic data suggested that the type 2IGF/mannose 6 phosphate receptor is an unlikely candidate for XR. Postnatal growth curves indicated that surviving Igf-1(-/-) mutants, which are infertile and exhibit delayed bone development, continue to grow with a retarded rate after birth in comparison with wild-type littermates and become 30% of normal weight as adults. PMID- 8402903 TI - Targeted disruption of IRF-1 or IRF-2 results in abnormal type I IFN gene induction and aberrant lymphocyte development. AB - Interferon regulatory factor 1 (IRF-1), a transcriptional activator, and its antagonistic repressor, IRF-2, were originally identified as regulators of the type I interferon (IFN) system. We have generated mice deficient in either IRF-1 or IRF-2 by gene targeting in embryonic stem cells. IRF-1-deficient fibroblasts lacked the normally observed type I IFN induction by poly(I):poly(C), while they induced type I IFN to similar levels as the wild type following Newcastle disease virus (NDV) infection. In contrast, IRF-2-deficient fibroblasts showed up regulated type I IFN induction by NDV infection. A profound reduction of TCR alpha beta+CD4-CD8+ T cells in IRF-1-deficient mice, with a thymocyte developmental defect, reveals a critical role for IRF-1 in T cell development. IRF-2-deficient mice exhibited bone marrow suppression of hematopoiesis and B lymphopoiesis and mortality following lymphocytic choriomeningitis virus infection. PMID- 8402904 TI - From fat yeast and nervous mice to brain myosin-V. PMID- 8402905 TI - Induction of muscle pioneers and floor plate is distinguished by the zebrafish no tail mutation. AB - Dorsal mesoderm is thought to provide important signals for axis formation and neural differentiation in vertebrate embryos. We have examined induction and patterning in a zebrafish mutant, no tail, that lacks a derivative of dorsal mesoderm, the notochord. Despite the absence of a differentiated notochord, development of the central nervous system including floor plate appears normal, likely owing to the presence of notochord precursor cells. In contrast, somites are misshapen, and muscle pioneer cells are absent. Wild-type cells transplanted into mutant hosts can autonomously differentiate into notochord and thereby rescue somitic defects, suggesting that interactions between notochord and paraxial mesoderm are necessary for proper somite patterning. Thus, cells derived from dorsal mesoderm may have multiple signaling functions during zebrafish embryogenesis. PMID- 8402906 TI - The signal for capacitative calcium entry. PMID- 8402907 TI - Chronic intestinal inflammation: an unexpected outcome in cytokine or T cell receptor mutant mice. PMID- 8402908 TI - Collapsin: a protein in brain that induces the collapse and paralysis of neuronal growth cones. AB - Repulsive guidance cues can steer neuronal growth cones during development and prevent mature axons from regenerating. We have identified a 100 kd glycoprotein in the chick brain that is a good candidate for a repulsive cue. Since it induces the collapse and paralysis of neuronal growth cones in vitro, we have named it collapsin. It is effective at concentrations of approximately 10 pM. The C terminal half of collapsin contains a single immunoglobulin-like domain and an additional highly basic region. The N-terminal half of collapsin shares significant homology with fasciclin IV, a growth cone guidance protein in grasshopper. Recombinant collapsin causes sensory ganglion growth cones to collapse but not retinal ganglion cell growth cones. We propose that collapsin could serve as a ligand that guides specific growth cones by a motility inhibiting mechanism. PMID- 8402909 TI - Bcl-2-deficient mice demonstrate fulminant lymphoid apoptosis, polycystic kidneys, and hypopigmented hair. AB - bcl-2-/-mice complete embryonic development, but display growth retardation and early mortality postnatally. Hematopoiesis including lymphocyte differentiation is initially normal, but thymus and spleen undergo massive apoptotic involution. Thymocytes require an apoptotic signal to manifest accelerated cell death. Renal failure results from severe polycystic kidney disease characterized by dilated proximal and distal tubular segments and hyperproliferation of epithelium and interstitium. bcl-2-/-mice turn gray with the second hair follicle cycle, implicating a defect in redox-regulated melanin synthesis. The abnormalities in these loss of function mice argue that Bcl-2 is a death repressor molecule functioning in an antioxidant pathway. PMID- 8402910 TI - Ulcerative colitis-like disease in mice with a disrupted interleukin-2 gene. AB - Mice deficient for interleukin-2 develop normally during the first 3-4 weeks of age. However, later on they become severely compromised, and about 50% of the animals die between 4 and 9 weeks after birth. Of the remaining mice, 100% develop an inflammatory bowel disease with striking clinical and histological similarity to ulcerative colitis in humans. The alterations of the immune system are characterized by a high number of activated T and B cells, elevated immunoglobulin secretion, anti-colon antibodies, and aberrant expression of class II major histocompatibility complex molecules. The data provide evidence for a primary role of the immune system in the etiology of ulcerative colitis and strongly suggest that the disease results from an abnormal immune response to a normal antigenic stimulus. PMID- 8402911 TI - Interleukin-10-deficient mice develop chronic enterocolitis. AB - Interleukin-10 (IL-10) affects the growth and differentiation of many hemopoietic cells in vitro; in particular, it is a potent suppressor of macrophage and T cell functions. In IL-10-deficient mice, generated by gene targeting, lymphocyte development and antibody responses are normal, but most animals are growth retarded and anemic and suffer from chronic enterocolitis. Alterations in intestine include extensive mucosal hyperplasia, inflammatory reactions, and aberrant expression of major histocompatibility complex class II molecules on epithelia. In contrast, mutants kept under specific pathogen-free conditions develop only a local inflammation limited to the proximal colon. These results indicate that the bowel inflammation in the mutants originates from uncontrolled immune responses stimulated by enteric antigens and that IL-10 is an essential immunoregulator in the intestinal tract. PMID- 8402912 TI - A novel disulfide-linked heterodimer on pre-T cells consists of the T cell receptor beta chain and a 33 kd glycoprotein. AB - We describe a novel signal-transducing protein complex, which consists of the T cell receptor (TCR) beta chain that is disulfide linked to a 33 kd glycoprotein and noncovalently associated with proteins of the CD3 complex on the surface of the pre-T cell line SCB.29. This 33 kd glycoprotein, provisionally designated gp33, represents neither of the known TCR chains and has escaped previous detection because it labels poorly by surface iodination. This glycoprotein is absent from the surface of mature T cell lines. A TCR beta complex with identical molecular masses before and after reduction can be immunoprecipitated from surface-iodinated large thymocytes of TCR alpha-deficient mice. The novel gp33 TCR beta complex may be entirely or partly responsible for control of early T cell development exerted by the TCR beta protein. PMID- 8402913 TI - The human CD46 molecule is a receptor for measles virus (Edmonston strain). AB - Measles virus normally causes disease in human beings, and the host range of this virus may be determined by a specific receptor on the surface of primate cells. Human-rodent somatic cell hybrids were tested for their ability to bind measles virus, and only cells that contained human chromosome 1 were capable of binding virus. A study of lymphocyte markers suggested that the complement regulator known either as membrane cofactor protein or CD46 was the measles virus receptor. We proved this hypothesis by demonstrating that hamster cell lines that expressed human CD46 could subsequently bind virus. Furthermore, infected CD46+ cells produced syncytia and viral proteins. Finally, polyclonal antisera against CD46 inhibited virus binding and infection. These results prove that human CD46 permits cells both to bind measles virus and to support infection. PMID- 8402914 TI - The Drosophila 78C early late puff contains E78, an ecdysone-inducible gene that encodes a novel member of the nuclear hormone receptor superfamily. AB - We report the molecular definition of an early late puff locus, at position 78C, that is inducible by ecdysone at the onset of Drosophila metamorphosis. This puff contains a single ecdysone-inducible gene consisting of two nested transcription units, E78A and E78B. E78A mRNA is expressed during a brief interval in mid-pupal development and encodes a novel member of the nuclear hormone receptor superfamily. E78B encodes a truncated receptor isoform that lacks the DNA-binding domain and is predominantly expressed at puparium formation and immediately following E78A in pupae. E78B is directly inducible by ecdysone in late third instar larvae and depends on ecdysone-induced protein synthesis for its maximal level of expression. These observations indicate that E78 represents a distinct subset of early ecdysone-inducible regulatory genes. PMID- 8402915 TI - The target of ammonia action in dictyostelium. AB - The weak base ammonia is implicated in a number of key processes in Dictyostelium development, notably aggregation and culmination. To determine its intracellular site of action, we compared its biological potency with that of other weak bases. All the bases inhibited these developmental processes effectively, but differed manyfold in potency in accordance with their activity in dissipating pH gradients, as measured by in vivo 31P NMR with pH-sensitive phosphonate probes. These results indicate that weak bases influence development by raising the pH of an intracellular acidic compartment. PMID- 8402916 TI - Translational regulation of tra-2 by its 3' untranslated region controls sexual identity in C. elegans. AB - C. elegans hermaphrodites make sperm and then oocytes in an otherwise female animal. Gain-of-function mutations in the sex-determining gene tra-2 (tra-2(gf)) transform hermaphrodites into females (spermless hermaphrodites). The tra-2(gf) mutations map to a perfect direct repeat in the 3' untranslated region; each repeat is called a direct repeat element (DRE). Three experiments demonstrate that DREs repress tra-2 at the translational level. First, tra-2(gf) mRNAs are associated with larger polysomes than are their wild-type counterparts. Second, translation of a reporter RNA is inhibited by DREs. Third, disruption of DREs does not increase tra-2 mRNA levels. An RNA binding activity specifically associates with the DREs. We propose that tra-2 translation is inhibited by association of an RNA binding-factor with the DREs and that this translational control is essential for development of C. elegans as a hermaphrodite/male species. PMID- 8402917 TI - Reverse branch migration of Holliday junctions by RecG protein: a new mechanism for resolution of intermediates in recombination and DNA repair. AB - The RecG protein of E. coli is a junction-specific DNA helicase involved in recombination and DNA repair. The function of the protein was investigated using an in vitro recombination reaction catalyzed by RecA. We show that RecG counters RecA-driven strand exchange by catalyzing branch migration of the Holliday junction in the reverse direction. This activity represents a new mechanism for resolving recombination intermediates that is independent of junction cleavage. We discuss how reverse branch migration can facilitate DNA repair, promote recombination in conjugational crosses, and confine the distribution of Chi stimulated cross-overs. We suggest that the RecG mechanism for resolution of junctions is universal and provides a simple system that allows gene conversion without associated crossing over. PMID- 8402918 TI - Two related recombinases are required for site-specific recombination at dif and cer in E. coli K12. AB - The stable inheritance of ColE1-related plasmids and the normal partition of the E. coli chromosome require the function of the Xer site-specific recombination system. We show that in addition to the XerC recombinase, whose function has already been implicated in this system, a second chromosomally encoded recombinase, XerD, is required. The XerC and XerD proteins show 37% identity and bind to separate halves of the recombination site. Both proteins act catalytically in the recombination reaction. Recombination site asymmetry and the requirement of two recombinases ensure that only correctly aligned sites are recombined. We predict that normal partition of most circular chromosomes requires the participation of site-specific recombination to convert any multimers (arising by homologous recombination) to monomers. PMID- 8402919 TI - bottleneck acts as a regulator of the microfilament network governing cellularization of the Drosophila embryo. AB - A dynamic network of cortical microfilaments is associated with the cleavage furrow membranes during cellularization of the Drosophila embryo. A specific set of structural rearrangements in this network is required for orchestration and execution of its mechanistic roles. We describe the characterization of the gene bottleneck (bnk), mutations in which disturb the proper sequence of rearrangements of the microfilament network, leading to a variety of morphological defects during cellularization. bnk, whose expression is restricted to the blastoderm stages of Drosophila embryogenesis, encodes a novel, exceptionally basic protein that specifically colocalizes with the microfilament network. The expression pattern and mutant phenotype of bnk suggest a direct role for this element in regulation of the dynamic restructuring of the actin-based cytoskeleton of cellularizing Drosophila embryos. PMID- 8402920 TI - An active tissue-specific enhancer and bound transcription factors existing in a precisely positioned nucleosomal array. AB - Nucleosomes positioned over promoters are usually inhibitory to protein binding and activity. We analyzed at the nucleotide level of resolution the nucleosomal organization of a distal, liver-specific enhancer in various mouse tissues and found that the enhancer exists in an array of three precisely positioned nucleosomes only in liver chromatin, where the enhancer is active. In vivo footprinting reveals that essential transcription factor-binding sites are occupied on apparent nucleosome surfaces, in one case leading to a perturbed nucleosomal structure. A similar nucleosomal array is generated with an in vitro chromatin assembly system in which nucleosome positioning is dependent upon binding to the enhancer of proteins related to hepatocyte nuclear factor 3. We suggest that certain transcription factors can organize nucleosomal structures that define an active enhancer element. PMID- 8402921 TI - Effects of rIL-7 on murine bone marrow NK precursor cells. AB - We recently demonstrated that functional NK cells are produced from their precursors in murine long-term bone marrow cultures without the addition of exogenous growth factors. Because IL-7 is known to be produced by bone marrow stromal cells and is a proliferation factor for some immature cells of the B and T cell lineages, we tested whether rIL-7 could support the proliferation or maturation of NK precursor cells. By itself, rIL-7 did not induce NK lytic activity in cultures of unseparated bone marrow cells (BMC), but it did augment the response of unseparated BMC to 50 U rIL-2/ml, with a maximal enhancement at 10 ng IL-7/ml. Depletion experiments demonstrated that the IL-7-induced increase in cytotoxicity was not due to NK precursors, however, but to mature T and NK cells. IL-7 preferentially increased the number of CD8+ cells. Preculture of NK 1.1-depleted BMC with IL-7 did not increase the total number of NK 1.1+ cells or the lytic activity generated from NK precursor cells. However, IL-2-responsive NK lineage cells survived better in IL-7-supplemented medium than in medium alone. Thus, soluble rIL-7 did not expand the NK precursor cell population in vitro but it maintained the viability and subsequent responsiveness to IL-2 of NK precursors. PMID- 8402922 TI - The biologic activity of ACTH and related peptides on peripheral blood mononuclear cells is altered by the presence of dexamethasone. AB - We wished to determine whether the glucocorticoid hormone dexamethasone (dex), a potent modular of both pituitary-adrenal and immunologic activity, altered the effect of adrenocorticotropin-related peptides on activated peripheral blood mononuclear cells (PBMCs) and T-cells. PBMCs preactivated with Concanavalin A (Con A) and T-cells preactivated with phorbol 12,13 dibutyrate (PDB) plus phytohemagglutinin (PHA) were exposed to dex, ACTH, and related peptide sequences. Proliferation of cells was measured as was IL-2 in conditioned media. It was found that in the absence of dex only the peptide ACTH(18-39) altered proliferation of PBMCs while there was no effect of peptide on T-cells activated via protein kinase C-mediated pathways. Significant reversal of the inhibitory effect of dex on proliferation of PBMCs exposed to Con A was achieved with addition of ACTH(1-39) and ACTH(11-24), while IL-2 levels were unaffected by the addition of peptide. ACTH(18-39) and ACTH(11-24) enhanced the inhibitory effect of dex on T-cells activated with PDB plus PHA. These findings suggest that the biologic activity of ACTH on immune cells is altered when dexamethasone is present and under certain circumstances ACTH may protect the immunologic response from the inhibitory effects of elevated ambient glucocorticoids. PMID- 8402923 TI - Reconstitution of lymphoid tissues under the influence of a subclinical level of graft versus host reaction induced by bone marrow T cells or splenic T cell subsets. AB - Reconstitution of lymphoid tissues under the influence of subclinical graft versus host reaction (GVHR) has been investigated. Lethally irradiated AKR mice were reconstituted with B10 bone marrow (BM) cells which had been treated with anti-Thy-1 antibody alone without complement (GVHR chimera). Their immunological reconstitution was analyzed and compared with that of AKR recipients which had been reconstituted with B10 BM cells treated with anti-Thy-1 antibody plus complement (control chimera). One hundred percent of both chimeras survived more than 100 days without showing clinical signs of GVHR. However, full donor chimerism was accomplished at an early stage after reconstitution in the former GVHR chimeras, whereas a substantial number of recipient T cells persisted in control chimeras for the entire observation period. When reconstitution of various lymphoid tissues was compared between control and GVHR chimeras, no difference in the reconstitution of the thymus and spleen was noted. By contrast, the cellularity of peripheral lymph nodes in GVHR chimeras was regularly considerably lower than that of the control chimeras. The apparent insufficiency of lymph node reconstitution appeared to be attributable to the impairment of lymph node structure itself which may be involved in lymphocyte homing. Furthermore, clonal deletion of V beta 6+ T cells which are reactive to recipient (Mls-1a) antigens was abrogated in the GVHR chimeras but was normally induced in the more completely T cell-depleted control chimeras. This abrogation of clonal deletion of V beta 6+ T cells appeared to result from the early disappearance of recipient T cells in these chimeras. Thus, it appeared that donor T cells in the BM that survive anti-Thy-1 treatment in vitro plus subsequent BM transplantation induced a subclinical level GVHR which contributed to the full donor chimerism as well as abrogation of clonal elimination of V beta 6+ donor T cells. Indeed, inoculation of CD8+ T cells along with the transplantation of the T cell-depleted BM cells (anti-Thy-1 plus C-treated cells) from donor mice into the AKR recipients was also shown to induce a similar state in the recipients. PMID- 8402924 TI - Effect of ethanol on development of fetal mouse thymocytes in organ culture. AB - Exposure of mouse fetuses to ethanol in utero retards thymus development. The direct effect of ethanol on growth and differentiation of thymocytes was studied using organ cultures of 14-day fetal mouse thymuses. Fetal thymus organ cultures containing 0.2 or 0.4% ethanol produced fewer total thymocytes, proportionately fewer CD4+CD8+ (immature) thymocytes, and proportionately more CD4+CD8- (mature) cells than untreated control cultures after 5 days of culture. Total cell numbers and proportions of CD4+CD8+ thymocytes declined in a dose-dependent manner with increasing ethanol concentrations from 0.2 to 0.8%. In time course studies, thymuses cultured with 0.4% ethanol had an increased percentage of CD4+CD8- cells at all days examined between Days 4 and 6. In the same experiments, thymuses exposed to ethanol underwent accelerated loss of the interleukin-2 receptor (a marker of immature prothymocytes) and had higher percentages of cells positive for the gamma delta-T-cell receptor. Exposure to ethanol for 16 to 20 hr increased the percentage of noncycling thymocytes. Furthermore, ethanol increased apoptosis in fetal thymocytes. Acetaldehyde, the immediate product of ethanol catabolism, had no effect on thymocyte subpopulation ratios or cell numbers at a physiologic concentration (50 microM). Results indicate that in a controlled in vitro model of thymus development, ethanol reduced cell numbers and altered proportions of thymocyte subsets defined by differentiation antigens. PMID- 8402925 TI - Distinct phenotypic and functional characteristics of human natural killer cells obtained by rapid interleukin 2-induced adherence to plastic. AB - Human natural killer (NK) cells can be functionally subdivided into adherent (A) and non-adherent (NA) subpopulations. In the presence of 22 nM of interleukin 2(IL2), a substantial proportion of resting (R)-NK cells developed adherence to plastic as early as after 5 min of IL2 incubation, and by 1-5 hr of IL2 induction, 16% (range, 4-30%) of NK cells were adherent. Optimal concentration of IL2 for adherence of NK cells was 2-22 nM. This adherence was blocked completely by antibody to IL2 receptor (IL2R)-beta and, partially, by antibodies to beta 1 or beta 2 integrins, ICAM-1, CD2 or LFA3, but not by antibodies to the IL2R-alpha or CD56 antigen. A-NK cells separated from NA-NK cells after 5 hr of incubation in the presence of IL2 were significantly (P < 0.05) enriched in CD56dimCD16dim or -IL2Rp55+ and IL2Rp75+ cells, but were depleted of CD56bright CD16- cells. While surface density of CD56 and CD16 antigens was lower, that of beta 2 integrins (CD18, CD11a, CD11b) was higher on A-NK than on NA-NK cells. In a single-cell cytotoxicity assay, 61% of A-NK vs 37% of NA-NK cells bound, and 24% of A-NK vs 11% of NA-NK cells killed, K562 targets. In 4-day cultures with 0.02 or 2.2 nM of IL2, A-NK cells developed lymphokine-activated killer (LAK) activity later than NA-NK cells. By autoradiography, three to eight times more A-NK than NA-NK cells incorporated [3H]TdR into cell nuclei between 48 and 96 hr of IL2 incubation. In 14-day cultures in the presence of 22 nM of IL2, A-NK cells, which were initially adherent but later grew as single-cell suspensions, proliferated better (30-fold; P < 0.03) and expressed lower membrane density of CD56 than NA NK cells. In culture, A-NK cells had consistently higher cytotoxicity against K562 targets than NA-NK cells, but cytotoxicity against Daudi was similar for both subsets. The data indicate that short incubation (1-5 hr) of human NK cells in the presence of 22 nM of IL2 allows for selection of a subpopulation which differs from the rest of NK cells not only by properties of rapid adherence to plastic, but also by a characteristic phenotype (CD3-CD56dim or -CD16dim or -beta 2integrinsbrightIL2Rp75+), rapid expression of IL2R-alpha, higher NK activity, delayed development of LAK activity, and ability to respond optimally in the presence of 22 nM of IL2.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402926 TI - MHC/peptide binding studies indicate hierarchy of anchor residues. AB - MHC class I molecules present octa- or nonapeptides derived from cellular proteins. Such peptides adhere to strict rules, which are individual to each MHC allele. Synthetic peptides conforming to these rules or peptides being at variance at critical residues were assayed for binding to MHC class I molecules. The binding assay employed the peptide-induced stabilization of MHC molecules of RMA-S cells. The data indicate that most proline-free peptides conforming to the allele-specific motifs of Kb or Db bind to the respective molecules, whereas peptides missing only one of the two allele-specific anchor residues lost their capacity to stabilize class I molecules on RMA-S cells. The residues allowed at anchor positions of the Kb motif are not equal in their binding efficiency and can be ordered in a hierarchic row. Residues at nonanchor positions may also influence efficiency of peptide binding or may require deviations from the standard peptide length. PMID- 8402927 TI - Identification and characterization of B cell precursors in rat lymphoid tissues. I. Adoptive transfer assays for precursors of TI-1, TI-2, and TD antigen-reactive B cells. AB - Quantitative adoptive transfer assays were developed to detect the precursors of TI-1, TI-2, and TD antigen-reactive B cells in rat lymphoid tissues. Studies on the immune responses in normal and athymic nude rats validate the use of TNP lipopolysaccharide as a TI-1 antigen, TNP-Ficoll as a TI-2 antigen, and SRBC as a TD antigen in rats. The precursors to these immunologically competent B cells are detected, following transfer into irradiated histocompatible recipients, by their ability to generate expanded populations of antigen-reactive B cells capable of mounting antibody responses (splenic IgM plaque-forming cells) to these antigens. Maximal numbers of antigen-reactive B cells emerge in antigenically naive rats after an interval of 7-12 days following transfer of donor lymphoid cells and decline rapidly thereafter. The delayed responses in adoptive recipients reconstituted with spleen cells are proportional to the numbers of spleen cells transferred and are shown to be primarily donor derived using histocompatible Ig kappa chain alloantigen disparate rat strain combinations. The precursors of TI 1, TI-2, and TD antigen-reactive B cells are present in both donor spleen and bone marrow. However, precursor cells to TI-1 and TD antigens are largely absent from donor lymph node cells, whereas precursors to the TI-2 antigen are as prevalent in donor lymph node as in donor spleen. These results support the hypothesis that newly formed virginal B cells represent transient populations of precursor cells that undergo further proliferation and differentiation in the spleen before acquiring immunological competence. The results also suggest that the precursors of TI-2 antigen-reactive B cells differ developmentally from those of TI-1 and TD antigen-reactive B cells, and that the antigen-reactive progeny of these precursors require additional stimulation in order to join the pool of long lived peripheral B cells. PMID- 8402928 TI - Occurrence of a soluble nonspecific suppressor factor in the serum early after birth. AB - A nonspecific suppressor factor has been identified in serum of newborn rats and calves. This factor, designated SUF-s, was shown to interfere--across species barriers--with lymphocyte responses in vitro and in vivo. SUF-s interferes in vitro with T- and B-cell proliferation induced by different mitogens and IL-2. Our findings indicate that the activity of SUF-s in vitro, which is of a reversible nature, is directed at an early event in the cascade of T-cell activation. SUF-s does not affect intrinsically regulated proliferation, such as that of tumor cells or established cell lines. In vivo, SUF-s prevents graft-vs host disease induced by transplantation of allogeneic bone marrow cells in lethally irradiated mice. Using of affinity chromatography, hydrophobic interaction chromatography, and gel filtration, a 15,000-fold purification of the suppressive factor was attained. The moiety engaged in suppression was identified as a 20- to 40-kDa protein. The biological activity is destroyed at temperatures above 70 degrees C, by proteolytic enzyme digestion and under alkaline conditions but was resistant to acidic and reducing conditions. Judged by its biological activity and some of its physical properties, SUF-s is most likely distinct from other described suppressor factors or known cytokines with suppressor activity, such as IL-4, IL-10, interferon-gamma, transforming growth factor-beta or alpha fetoprotein. PMID- 8402929 TI - The host response in graft versus host disease. I. Radiosensitive T cells of host origin inhibit parental anti-F1 cytotoxicity in murine chronic graft versus host disease. AB - Murine graft versus host (GVH) disease takes two forms depending on the parental/F1 strain combination employed. Acute lethal GVH disease is characterized by anemia, lymphopenia, hypogammaglobulinemia, profound anti-F1 cytotoxicity, and the loss of cytotoxic potential against third-party alloantigen. In contrast to this, chronic GVH disease is characterized by polyclonal B cell activation, auto-antibody production, no anti-F1 cytotoxicity, and retained cytotoxicity against allotargets. We now report that this marked disparity in disease expression results from a radio-sensitive host mechanism which protects the F1 mouse from parental anti-F1 cytotoxicity in mice undergoing chronic GVH disease. Cellular analysis revealed that protection in chronic GVH disease is mediated by a phenotypically complex system of genetically unrestricted radiosensitive T cells of F1 origin. These cells fail to functionally emerge in mice undergoing acute lethal GVH disease. PMID- 8402930 TI - The host response in graft versus host disease. II. The emergence of host protective cells is in part determined by background genomic compatibility. AB - Murine graft versus host (GVH) disease takes two forms depending on the parental/F1 strain combination employed. In an accompanying paper (Singh et al., Clin. Immunol. 151, 1993) many of the clinical features of these two forms of GVH disease are described. In addition to these clinical characteristics, acute lethal GVH (ALGVH) disease is characterized by diminished natural killer cell activity, whereas chronic GVH disease is characterized by normal or increased natural killer cell activity. Previously we have reported that this marked disparity in disease expression can be attributed to radiosensitive host cells which protect the F1 mouse from parental anti-F1 cytotoxicity (CTX) in mice undergoing chronic GVH (CGVH) disease. These cells fail to functionally emerge in mice undergoing ALGVH disease. We now report that the background genome, presumably the minor lymphocyte stimulatory loci, of the donor cells determines whether these host cells emerge and thereby dictates the form of GVH disease which is induced. C57BL/6 (B6) cells (H-2b, minor lymphocyte stimulatory locus (Mls)b) and B10.D2 cells (H-2d, Mlsb) were found to induce ALGVH disease when adoptively transferred to [C57BL/6xDBA/2]F1 (B6D2) (H-2b/d, Mls-1a/b, Mls-2a/b) recipient mice. DBA/2 cells (H-2d, Mls-1a, Mls-2a) and Balb/c cells (H-2d, Mls 1a, Mls-2b) induced CGVH disease in B6D2 mice. Using Mls congenic strains we have demonstrated that donor cell reactivity against Mls-2a was necessary and sufficient to induce ALGVH disease as determined by anemia, lymphopenia, anti-F1 cytotoxicity, and loss of cytotoxicity against allogeneic targets. Such Mls-2a reactivity correlated with the impaired induction of a host protective cell capable of vetoing self-directed CTX. Failure of this host protective cell to emerge in turn correlated with donor anti-host CTX and the emergence of ALGVH disease. PMID- 8402931 TI - The secondary antigen-specific IgE response in murine lymphocytes is resistant to blockade by anti-IL4 antibody and an antisense oligodeoxynucleotide for IL4 mRNA. AB - In order to investigate the role of interleukin 4 (IL4) in the induction of antigen-specific IgE responses, we established culture conditions which allow the induction of anti-trinitrophenyl(TNP) IgE response by the coculture of TNP keyhole limpet hemocyanin-primed C3H B cells with conalbumin (CA)-specific type 2 helper T (Th2) cell clone, D10.G4.1 in the presence of TNP-CA. A maximum level of anti-TNP IgE was secreted at 1 microgram/ml TNP-CA. By using filter-separated double-chamber culture plates, the physical contact between T cells and B cells was shown to be necessary in this response. Anti-TNP IgE synthesis was not significantly suppressed in the presence of 20-40 micrograms/ml monoclonal anti IL4 (11B11), nor was the response further enhanced by the addition of recombinant IL4. 11B11 added together with anti-IL5 had also no suppressive effects on the IgE response. This apparent independence on IL4 might be due to the fact that IL4 is transferred from T cells to B cells in a transsynaptical way that would be refractory to the neutralization by 11B11. In order to test this possibility, we synthesized the phosphorothioate analogue of an antisense oligodeoxynucleotide against IL4 mRNA (S-oligo) for inhibiting IL4 production from Th2 cells specifically. S-oligo was effective at 10-20 micrograms/ml in suppressing IL4 production from D10.G4.1 cells by 80-90%. It was demonstrated that S-oligo, either alone or in combination with 11B11, did not significantly suppress anti TNP IgE response. These results suggest that antigen-specific IgE response in primed B cells does not depend on IL4, but requires cognate interaction with Th2 cells. PMID- 8402932 TI - Cells recovered from human DTH reactions: phenotypic and functional analysis. AB - Using the skin window chamber (SC) technique T cells were recovered from DTH reactions elicited by either PPD or Candida (CAN) in three healthy individuals. Analysis of representative panels of clones established in limiting dilution cultures from SC and blood disclosed a 2.6-, 10-, and 314-fold higher incidence of antigen-specific T cells in SC than in blood. Thirty CD4+ and 13 CD8+ antigen specific clones were recovered from SC. Eight of 14 CD4+ clones displayed lectin mediated cytotoxicity (as compared to 2/10 from blood) and 4/5 CD8+ clones were also cytotoxic. The CD4+ clones from SC, but not from blood could be divided into two groups, one producing IFN-gamma and IL-2, and the other providing help for Ig synthesis. Among the CD8+ SC clones analyzed the noncytotoxic clone could provide help for Ig synthesis but produced no IFN-gamma, whereas the other four showed the opposite pattern. Analysis of the fluid phase of SCs from positive DTH reactions disclosed high levels of TNF and IFN-gamma; neither was detected from SC's covering sites injected with nonspecific antigens. PMID- 8402933 TI - Induction of CD45 isoform switch in murine B cells by antigen receptor stimulation and by phorbol myristate acetate and ionomycin. AB - In this study, we examined whether CD45 isoform can be switched in murine mature B cells and what signals are responsible for the process. Stimulation of murine splenic B cells with lipopolysaccharide did not reduce the expression of CD45RA-, B-, and C-exon-dependent epitopes or a CD45 common epitope, but rather enhanced the expression. Stimulation with goat antimouse IgM antibody did not significantly reduce CD45 expression but caused a partial reduction in the expression of CD45RA-, B-, and C-exon-dependent epitopes. Phorbol myristate acetate (PMA) alone did not significantly alter the expression of CD45 but the combination of PMA and ionomycin induced a strong reduction in the expression of CD45RA-, B-, and C-exon-dependent epitopes without affecting the level of CD45 common epitope expression. Reverse transcription and polymerase chain reaction analysis demonstrated that CD45 isoform switch induced by anti-IgM or PMA plus ionomycin is indeed mediated by alternative splicing of A-, B-, and C-exon derived mRNA. These results suggest that CD45 isoform of murine mature B cells can be switched by antigen receptor-mediated signals, and the process seems to be regulated at least in part by protein kinase C activation and mobilization of calcium ions. PMID- 8402934 TI - Regulation of human IgE response in hu-PBL-SCID mice. AB - The human IgE response was investigated in hu-PBL-SCID mice created by ip injection of human PBL into C.B.17 scid/scid (scid) mice. With 30-100 x 10(6) PBL/mouse, 80 to 90% of the animals responded with human IgE serum levels of 3 1000 ng/ml after 2 weeks. PBL from all donors analyzed (total number > 20) responded with IgE production. The half-lives of human IgE, IgM, and IgG in scid mice were determined to test the possibility of a passive transfer of the immunoglobulins in contrast to de novo synthesis. The values found were 88, 128, and 126 hr, respectively. In general, immunoglobulin production of all isotypes continuously increased over a period of 7-9 weeks after PBL injection, indicating de novo synthesis had taken place. The kinetics of the IgE response exhibited two phases: An initial burst of IgE production occurred between Days 12 and 22. This burst reached levels of 25-70 ng/ml IgE. After a rapid decline to about 50% of the peak value there was a sustained, slow, increase of IgE production for several weeks, excluding a passive transfer for IgE. About half of the donors lacked the initial burst of IgE production and only exhibited a slowly rising IgE production that is indistinguishable from the slow phase of the former donor population. The levels reached in this second phase of IgE production were 20-40 ng/ml after 6-7 weeks. This kinetics may reflect the presence of two different B cell populations, of which only one is present in all donors. The initial IgE burst was only partially dependent on the presence of human IL-4, reflected by a partial inhibition of this response by a neutralizing monoclonal anti-IL-4 antibody. The IgE response in scid mice seems to consist therefore of an IL-4 independent and an IL-4-dependent part, indicating the response to be partially driven by preswitched B cells. Injection of exogenous recombinant human IL-4 (rhIL-4) was not suitable due to the short half-life of rhIL-4 in scid mice of 12 min. Attempts to supply a constant source of rhIL-4 by injection of IL-4 producing Chinese hamster ovary cells failed because of toxic effects produced by these cells. The human IgE production in the scid mice was suppressed by interferon-alpha (BD) to 60-80% compared to that of untreated mice. The suppression was not isotype specific, however, because production of IgG and IgM was inhibited to similar extents.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8402935 TI - Pivotal role of endogenous TNF-alpha in the IL-2-driven activation and proliferation of the functionally immature NK free subset. AB - Highly purified peripheral blood-derived NK cells can be separated into three functionally defined subpopulations, namely, non-target binding free cells, binders, and killers. The free cell subset is the least mature and was examined for its response to IL-2-mediated activation and the role of endogenous secretion of TNF-alpha in its maturation and differentiation. The findings demonstrate that endogenous TNF-alpha secretion is prerequisite for the initiation of IL-2 mediated activation of free cells into killer cells. The addition of IL-2 to free cells upregulated the surface expression of IL-2R (TAC), TNF-R (p75), CD69, and ICAM-1 antigens and also stimulated cell proliferation. Furthermore, the addition of IL-2 to free cells resulted in the induction of cytotoxic activity and stimulation of free cells to become binder and killer cells. All of these IL-2 mediated manifestations are shown to be downregulated by the addition of anti-TNF alpha antibody. However, IL-2-mediated TNF-alpha secretion was not affected by the addition of anti-TNF-alpha antibody. The specificity of the anti-TNF-alpha antibody-mediated inhibition was corroborated by the failure of the antibody to inhibit interferon-alpha-mediated activation of free cells into killer cells. Most of the events associated with the inhibitory activity of anti-TNF-alpha antibody were mimicked by the addition of IL-4 to IL-2-treated free cells. These findings suggest that the IL-2-mediated maturation and differentiation of the immature free cells to become cytotoxic and proliferate are the result of a sequence of events that are initiated by the secretion of endogenous TNF-alpha. PMID- 8402936 TI - Carcinogen-treated skin allografts rejected by T lymphocytes specific for class I but not class II MHC antigens. AB - Treatment of skin with the chemical carcinogen 7,12-dimethylbenz[a]anthracene (DMBA), which reduces the density of epidermal class II MHC-expressing Langerhans cells (LC), enhances its survival when transplanted onto histoincompatible hosts. We have examined the ability of T lymphocytes which reject DMBA-treated skin to lyse P388D1 cells expressing either only class I or class I and II antigens. Lymphocytes isolated from solvent-treated grafts showed greater cytotoxicity for the targets expressing both antigens, indicating that some of these lymphocytes were specific for class II MHC antigens. In contrast, lymphocytes isolated from carcinogen-treated grafts lysed both targets similarly and hence did not contain any cells specific for class II MHC antigens. Anti-class I MHC antibody blocked cytotoxicity by both leukocyte populations to similar extents, but anti-class II MHC antibodies preferentially blocked T cells isolated from the solvent-treated grafts. There was no difference in the phenotype of the cytotoxic cells isolated from solvent- and carcinogen-treated grafts. Thus, whereas solvent-treated skin grafts are rejected by T cells specific for class I and II MHC antigens, DMBA treated skin grafts are only rejected by class I MHC-specific T cells which may account for the enhanced survival of the carcinogen-treated grafts. PMID- 8402937 TI - Cytokine production in first trimester chorionic villi: detection of mRNAs and protein products in situ. AB - Growth factor cytokines must play important roles in placental growth and differentiation. We used cDNA-mRNA in situ hybridization to examine whether human first trimester chorionic villi could produce these substances locally. IL-1 beta and IL-4 mRNA was found in both trophoblast layers, whereas CSF-1 and TNF alpha were found mostly in syncytiotrophoblast with much less localization to cytotrophoblast areas. IFN-gamma message was exclusively found in the cytotrophoblast. Immunohistochemical analysis confirmed that cytokines are produced by placental tissue, and the patterns of expression correlated well with the data obtained by in situ hybridization. These data are discussed regarding possible roles of cytokines in regulation of MHC antigen expression by trophoblasts and immunomodulation of decidual lymphocytes. PMID- 8402938 TI - Macrophage control of Brucella abortus: role of reactive oxygen intermediates and nitric oxide. AB - Macrophages infected with Brucella abortus are able to kill intracellular brucellae over the first 12 to 24 hr following infection. Thereafter, the surviving brucellae replicate. We have shown previously that macrophages activated with interferon-gamma (IFN-gamma) have enhanced brucellacidal and brucellastatic activities. In studies reported here, treatment of macrophages with methylene blue, an electron carrier, enhanced the initial killing of intracellular brucellae, indicating their susceptibility to reactive oxygen intermediates. In addition, inhibitors of reactive oxygen intermediates partially blocked the antibrucella activities exhibited by both non-cytokine-treated and IFN-gamma-activated macrophages. In contrast, addition of up to 5 mM NG monomethyl-L-arginine, to block generation of nitric oxide, resulted in minor but significant levels of blocking of macrophage antibrucella activities only when macrophages were not maximally activated even though maximally activated macrophages produced nitric oxide as indicated by accumulation of nitrite in culture supernatants. In addition, while the J774A.1 macrophage cell line had antibrucella activities which were enhanced by IFN-gamma activation, it did not produce nitric oxide when activated with IFN-gamma and infected with B. abortus. Finally, the IFN-gamma-induced enhancement of antibrucella activities by peritoneal macrophages was inhibited by addition of antitumor necrosis factor alpha (TNF-alpha) antibodies to the cultures, indicating that TNF-alpha is necessary for full expression of the macrophage antibrucella activities. PMID- 8402939 TI - The mechanism of epipodophyllotoxin-induced thymocyte apoptosis: possible role of a novel Ca(2+)-independent protein kinase. AB - The epipodophyllotoxins, etoposide (VP-16) and teniposide (VM-26), inhibit topoisomerase II activity by stabilization of the cleavable complex between the enzyme and DNA and formation of protein-bound double-stranded DNA breaks. While it is thought that these agents are cytotoxic by preventing cells from completing the S phase or undergoing mitosis, recent evidence suggests that these agents are also potent inducers of programmed cell death or apoptosis in both normal and malignant cells. We have examined the intracellular pathway leading to epipodophyllotoxin-induced apoptosis in normal mouse thymocytes. Epipodophyllotoxin-induced apoptosis may proceed via a mechanism that is independent of inhibition of topoisomerase activity per se because novobiocin and coumermycin, which inhibit the ATPase subunit of topoisomerase II, were relatively inefficient inducers of apoptosis in these cells, under conditions where strong apoptosis by the epipodophyllotoxins and dexamethasone could be observed. In addition, camptothecin, which inhibits topoisomerase I by stabilization of the cleavable complex between that enzyme and DNA, was also a poor inducer of apoptosis in these cells. Our data suggest that epipodophyllotoxin-induced mouse thymocyte apoptosis, like that induced by dexamethasone, proceeds via a mechanism that involves protein kinase C (PKC) or a similar enzyme. Apoptosis induced by VM-26 or by dexamethasone was inhibited by 1 (5-isoquinolinylsulfonyl)-2- methylpiperazine dihydrochloride (H7), an inhibitor of both PKC and cAMP-dependent protein kinases, but was relatively unaffected by N-(2-guanidinoethyl)-5-isoquinolinesulfonamide (HA1004), a more specific inhibitor of cAMP-dependent protein kinases. A more specific inhibitor of PKC, sangivamycin, also inhibited both VM-26-induced and dexamethasone-induced apoptosis. Both VM-26- and dexamethasone-induced apoptosis were unaffected by EGTA, a calcium (Ca2+) chelator, under conditions that inhibited apoptosis induced by the Ca2+ ionophore A23187. Moreover, while strong increases in intracellular Ca2+ were observed in thymocytes treated with A23187, we failed to detect increases in intracellular Ca2+ in cells induced to apoptose with either VM-26 or dexamethasone within the first 2 hr of culture. These results suggest that in mouse thymocytes there are at least two intracellular pathways leading to apoptosis: one, utilized by glucocorticoid and the epipodophyllotoxins, that proceeds in the absence of detectable increases in intracellular Ca2+ and possibly requires a novel Ca(2+)-independent PKC-like enzyme and another, utilized by Ca2+ ionophores, that is at least partially dependent on increased intracellular Ca2+. PMID- 8402940 TI - Tumor necrosis factor (TNF alpha) regulates intestinal mucus production during salmonellosis. AB - Mucus production by goblet cells in the gastrointestinal tract following Salmonella typhimurium infection using a ligated ileal loop model in mice was investigated. Assessment of the morphology of the loop tissue after Salmonella challenge revealed generalized tissue inflammation, characterized by edema and an inflammatory cell infiltrate. Villi were shortened and blunted, and crypts contained an increased number of cells with mitotic figures. Production of TNF alpha in the loops followed Salmonella challenge and occurred at the same time as the pathological sequelae. A nearly 50% decrease in the number of goblet cells in infected tissue compared to tissue from noninfected controls was observed at these same times. The sulfation of mucins produced by the goblet cells in infected tissues was increased in the villi but was unchanged in the crypts compared to uninfected tissues. Treatment of mice with antibody to TNF alpha before Salmonella challenge abrogated tissue pathology and returned goblet cell numbers and mucin profiles to those observed in noninfected controls. Our results indicate that TNF alpha may mediate changes in goblet cell expression and mucin sulfation in response to Salmonella challenge. PMID- 8402941 TI - Upregulation of T cell receptor gamma chain transcription by interleukin-2. AB - Signals involved in TcR gene rearrangement and expression are poorly understood. The ability of interleukin 2 to control TcR gamma gene expression was examined. Precursor thymocytes grown in the presence of IL-1 and Con A develop to CD3+ T cells that express either the TcR alpha/beta or gamma/delta complex on the cell surface (1). We show here that a major subpopulation of these cells are CD4-/CD8 , gamma/delta cells expressing only low levels of TcR gamma transcripts, compared to TcR delta mRnA or to TcR gamma mRNA of gamma/delta cells grown from precursor thymocytes with IL-2 and Con A. The cells cultured with IL-1 and lectin then strongly reacted to IL-2 by upregulating the steady-state level of TcR gamma mRNA. Our findings indicate that IL-2 upregulates TcR gamma transcription by transcriptional regulatory effects in this in vitro system. PMID- 8402942 TI - Impaired bone marrow-derived macrophage differentiation in vitamin D deficiency. AB - The calcium-regulating hormone, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), is also recognized as an immunomodulator. In vitro addition of the hormone to bone marrow derived macrophages (BMDMs) results in a decreased proliferation and an increased differentiation. In the present work we compare the in vitro differentiation of BMDMs derived from vitamin D-depleted and -repleted mice (-D and +D BMDMs, respectively). -D BMDMs proliferate in vitro slower than +D BMDMs. Addition of the hormone to BMDM cultures inhibited the rate of their proliferation, which was more pronounced in low-density cultures. The ability of mononuclear phagocytes to produce reactive oxygen metabolites is an important element in the microbicidal functions of these cells. We found that -D BMDMs produce less H2O2 than +D BMDMs, which was corrected by the in vitro addition of 1,25(OH)2D3. Analyses of various macrophage-specific surface antigens revealed a reduction in their expression on D BMDMs. In vitro addition of 1,25(OH)2D3 to BMDM cultures increased the expression of these antigens. The activity of the lysosomal enzyme acid phosphatase was similarly affected by vitamin D deficiency and by the in vitro addition of the hormone. Thus, vitamin D deficiency is associated with impaired maturation of BMDMs suggesting that the hormone is a natural modulator of macrophage maturation. PMID- 8402943 TI - Variants of vasoactive intestinal peptide in mouse mast cells and rat basophilic leukemia cells. AB - Radioimmunoassays for neuroendocrine vasoactive intestinal peptide (VIP1-28) detected 30-120 fmol of structurally related peptides in extracts of 10(7) mouse peritoneal mast cells, bone marrow-derived mast cells, cultured PT-18 and C1.MC/C57.1 lines of mast cells, and rat basophilic leukemia (RBL) cells. No VIP was found in peritoneal cells of mast cell-deficient WBB6F1-W/Wv mice, whereas the amounts extracted from peritoneal cells of the congenic normal (WBB6F1-+/+) mice were similar to those from cultured mouse mast cells. Sephadex G-25 gel filtration resolved two different-sized variants of VIP from mouse mast cells and RBL cells. Amino acid sequence analyses showed that the smaller variant is VIP10 28. The principal amino-terminally larger variant of VIP from C1.MC/C57.1 mouse mast cells and RBL cells exhibited amino acid sequence homology with VIP(-6)-28, and this sequence was established for the corresponding larger VIP from PT-18 mast cells. Polymerase chain reaction amplification of two different substituent sequences of prepro VIP in RBL cell RNA identified the VIP message. VIP10-28 was released from mouse mast cells concurrently with histamine by IgE-dependent stimulation. Rodent mast cell-derived VIP thus consists of both the truncated VIP10-28 and amino-terminally larger forms that appear to be generated by peptidolysis of a preproVIP similar to that found in neural cells. PMID- 8402944 TI - Characterization of T and B cells isolated from mucosa-associated tissues of the rhesus macaque. AB - This study has assessed T cell subsets according to the expression of CD4 and CD8 and the isotype of surface Ig+ (sIg+) B cells and plasma cells in mononuclear cells (MC) isolated from mucosa-associated tissues of rhesus macaques in comparison to lymphoid cells from systemic tissues, i.e., spleen and peripheral lymph nodes (PLN). Using enzymatic and/or mechanical dissociation methods, mononuclear cells were isolated from lamina propria (LP) of the small (jejunum and ileum) and large (cecum) intestines, parotid glands (PG), and mesenteric lymph nodes (MLN). Approximately equal numbers of CD4+ and CD8+ T cells occurred in mucosa-associated tissues (CD4:CD8 ratios of 0.9-1.2), while slightly higher numbers of CD8+ T cells were seen in spleen and PLN (CD4:CD8 ratios of 0.6-0.9). When the isotypes of sIg+ B cells were assessed in MC isolated from mucosa associated tissues, the highest frequency of sIgA+ B cells was seen in MLN, which may represent B cells which had recently migrated from IgA inductive sites. However, IgA effector tissues, such as intestinal LP and PG, contained frequencies of sIgA+ B cells lower than those of the MLN. When Ig-producing cells were examined, 50-90% of Ig-secreting cells were of the IgA class. This suggests that sIgA+ B cells differentiate into IgA plasma cells at higher rates in the mucosal effector tissues. Plasma cells of IgM isotype were also found in significant numbers in the LP of the intestine and PG, while IgG plasma cells were most prevalent in spleen and PLN. Taken together, mucosa-associated tissues of rhesus macaques are characterized by higher numbers of CD4+ T cells and Ig secreting plasma cells of IgA and IgM isotypes than those of systemic lymphoid tissues. PMID- 8402945 TI - Regulatory effect of interferon-gamma and phorbol esters on the surface expression and biosynthesis of MHC class I antigens by human leukemia cells. AB - We have used cell surface radioiodination, biosynthetic incorporation of [35S]methionine, and flow cytometry to analyze the effects of interferon gamma (IFN-gamma) and/or phorbol esters (PMA) on the turnover and expression of class I antigens of a human leukemia B cell line. Our results demonstrated that although both IFN-gamma and PMA enhance HLA expression, they act synergistically to increase by eightfold the amount of HLA polypeptides synthesized by the acute lymphoblastic leukemia cells and acted additively to augment the cell surface expression of HLA as quantified by flow cytometry. We observed a cyclic increase or decrease in the expression of class I antigens as a function of time in cell culture. IFN-gamma and/or PMA modulated this effect inducing more cells to express HLA maximally. These results suggest that there is a physiologic limit for the expression of major histocompatibility complex class I antigens. PMID- 8402946 TI - Effects of combinations of cytokines on murine antigen-specific B lymphocytes. AB - Cytokines are among the most important mediators in the immune response. They act on B lymphocytes at various stages along the activation pathway. To study the effects of combinations of cytokines, we have used an antigen-specific single B cell-cloning system devised in this laboratory. We report here that IL-1 can enhance the proliferative response and IgM secretion by B cells induced by IL-6. The results of a similar study demonstrated that IL-1 can also enhance the effects of IL-5 on B cell proliferation and IgM secretion in an additive manner. Kinetic analyses showed that IL-1 had to be present from the beginning of the culture for an optimum cooperative effect, whereas the addition of IL-6 could be delayed up to 2 days without a significant reduction of the response. In contrast, IL-5 had to be added together with IL-1 at the onset of culture to promote an optimum response. The responses elicited by IL-1 plus IL-6, IL-1 plus IL-5, and IL-4 plus IL-5 were almost identical. The addition of further cytokines to these cultures gave no enhancement above the effects observed with the two cytokine combinations. PMID- 8402947 TI - Glucocorticoid hormones upregulate interleukin 2 receptor alpha gene expression. AB - We present evidence that glucocorticoid hormones increase expression of IL-2Rec alpha chain on T cells by regulating IL-2Rec alpha gene transcription. We have previously reported that glucocorticoids can upregulate IL-2Rec alpha mRNA and protein expression in some T cell hybrids. In the present study we show that the glucocorticoid analogue dexamethasone increases mRNA levels of the endogenous IL 2Rec alpha gene and the expression of plasmids containing 5'-flanking sequences of the IL-2Rec alpha gene linked to CAT reporter genes transiently transfected into different cell lines. We show that the dexamethasone effect depends on cis acting regulatory elements in a segment (-1835/-802) of the mouse gene that also contains cytokine response elements and a inducible DNase I-hypersensitive site. Dexamethasone responses of IL-2Rec alpha-CAT reporter gene constructs were observed in a CTL line, in an IL-3-dependent bone marrow-derived cell line, and in COS7 monkey kidney cells. In the latter the response depended on cotransfection of a glucocorticoid receptor expression vector. The biological relevance of the glucocorticoid-mediated upregulation of the IL-2Rec alpha gene is discussed. PMID- 8402948 TI - Interleukin-2 production by T cells: a study of the immunoregulatory actions of interferon-alpha, interferon-gamma, and tumor necrosis factor-alpha in phenotypically different T cell clones. AB - The production of interleukin 2 (IL-2) by phytohemagglutinin (PHA)-stimulated human leukemia T cell lines MOLT-13 and MOLT-14, the T cell receptor (TcR) gamma delta-bearing clones, was significantly increased by adding human natural interferon-gamma (IFN-gamma) to the cell cultures. In addition, the IL-2 production by PHA-stimulated MOLT-13 and MOLT-14 cells was also augmented by human natural tumor necrosis factor-alpha (TNF-alpha) in a dose-dependent manner but was inhibited by IFN-alpha even at low concentrations (10 IU/ml). This pattern of reactivity is different from the responsiveness of the MOLT-16 cells, a TcR alpha beta-bearing cell line, which respond to IFN-alpha and TNF-alpha, but not to IFN-gamma, by augmentation of IL-2 production. These results suggest that (1) the addition of IFN-gamma has a direct effect on PHA-stimulated MOLT-13 and MOLT-14, resulting in an augmentation of IL-2 production; (2) phenotypically different T cell lines vary in their susceptibility to IFN-alpha and IFN-gamma regulation; and (3) the augmentation of IL-2 production by TNF-alpha appears to have no correlation with phenotypically different T cell clones. PMID- 8402949 TI - Comparison of the sensitivity of in vivo and in vitro assays for detection of antiviral cytotoxic T cell activity. AB - Sensitivities of in vitro and in vivo assays for the detection of lymphocytic choriomeningitis virus (LCMV)-specific CTL were compared. Measurement of primary cytotoxicity was the least sensitive of all the tested assays. However, when the same 51Cr release assays were performed after in vitro restimulation, this in vitro method was found to be more sensitive than all of the five in vivo assays tested. Assessment of CTL-mediated protection against LCMV replication in spleens was most sensitive among the in vivo tests, followed by CTL-mediated resistance to intracerebral or intraperitoneal challenge infections with vaccinia-LCMV recombinant virus. Inhibition of LCMV-induced choriomeningitis and of the footpad swelling reaction were least sensitive in detecting LCMV-specific CTL. As discussed, the presented sensitivity gradient may most probably be generalized. PMID- 8402950 TI - Immobilized staphylococcal enterotoxin A is sufficient to induce T cell proliferation. AB - We investigated the role of HLA-DR molecules in T cell stimulation by staphylococcal enterotoxin A (SEA). Previous results with immobilized purified HLA-DR preincubated with peptide showed that peptide-specific T cell clones were able to bind to and proliferate in response to purified HLA-DR/peptide complexes in the absence of antigen presenting cells. We report here that two human T cell clones (1 alpha beta and 1 gamma delta T cell clone) and a murine T cell hybridoma were each activated by immobilized purified HLA-DR4Dw4 preincubated with SEA. Furthermore, immobilized SEA in the absence of HLA-DR4Dw4 also stimulated the human T cell clones. The proliferative response of the human T cell clones was inhibited by CD3-reactive monoclonal antibodies, indicating that the T cell receptor (TCR)/CD3 reacts with SEA. These observations suggest that the HLA-DR in the complex functions only to immobilize SEA and that an interaction between the TCR and HLA-DR is not necessary for SEA-driven T cell stimulation. Finally, the assays described here could provide a method for defining and distinguishing the SEA binding sites for MHC class II and TCR. PMID- 8402951 TI - Anesthesia inhibits interferon-induced natural killer cell cytotoxicity via induction of CD8+ suppressor cells. AB - Anesthesia (Avertin, halothane, isoflurane, ether, or ketamine/xylazan) of mice inhibits subsequent stimulation of splenic natural killer cell (NK) cytotoxicity by interferon (IFN) treatment either in vitro and in vivo. The current data demonstrate (a) in vitro depletion of CD8+ cells from mononuclear splenocytes of anesthetized mice restored the ability of NK cells to respond in vitro to IFN stimulation and (b) coincubation of CD8+ splenocytes from anesthetized mice with CD8- splenocytes of naive mice resulted in a significant reduction of the IFN induced stimulation of NK activity in the coculture. These results suggest that anesthesia induces CD8+ cells that suppress stimulation of NK cytotoxicity by IFN. PMID- 8402952 TI - Recent quantitative studies of actin filament turnover during cell locomotion. AB - Cell locomotion depends on polymerization and depolymerization of filamentous actin. Net polymerization at the cell front occurs fast enough to fill the extending lamellipod, and since total F-actin is essentially constant over time, depolymerization must equal polymerization. Indeed, the fastest moving cell types have the highest rates of depolymerization. Accounting for the high rate of depolymerization raises several problems. One is that net depolymerization requires the concentration of G-actin to be low (below the critical concentration), but rapid polymerization (occurring < 1 micron away) requires the concentration of G-actin to be high (well above the critical concentration). This may be accomplished by spatial compartmentalization of factors that favor polymerization or depolymerization, and/or by proteins that bind G-actin and prevent spontaneous polymerization while allowing barbed-end elongation. A second problem is that depolymerization proceeds faster than would seem possible from studies of F-actin in vitro (as calculated from number and lengths of filaments present and in vitro rate constants). Rapid depolymerization may be accomplished by filament cutters or by cytoplasmic components (as yet undiscovered) that increase the rate of depolymerization. PMID- 8402953 TI - Localization of CapZ during myofibrillogenesis in cultured chicken muscle. AB - Actin filaments undergo dramatic changes in their organization during myofibrillogenesis. In mature skeletal muscle, both CapZ and the barbed end of the actin filaments are located at Z-discs. In vitro, CapZ binds the barbed end of actin filaments and prevents actin subunit addition and loss; CapZ also nucleates actin polymerization in vitro. Taken together, these properties suggest that CapZ may function to organize actin filaments during myofibrillogenesis. We report here that the amount of CapZ in myofibrils from adult chicken pectoral muscle is sufficient to "cap" each actin filament of the sacromere. Double immunofluorescence microscopy of skeletal muscle cells in culture was used to determine the spatial and temporal distributions of CapZ relative to actin, alpha actinin, titin, and myosin during myofibrillogenesis. Of particular interest was the assembly of CapZ at nascent Z-discs in relation to the organization of actin filaments in nascent myofibrils. In myoblasts and young myotubes, CapZ was diffusely distributed in the cytoplasm. As myotubes matured, CapZ was initially observed in a uniform distribution along non-striated actin filaments called stress fiber-like structures (SFLS). CapZ was observed in a periodic pattern characteristic of mature Z-discs along the SFLS prior to the appearance of a striated staining pattern for actin. In older myotubes, when actin was observed in a pattern characteristic of I-bands, CapZ was distributed in a periodic pattern characteristic of mature Z-discs. The finding that CapZ was assembled at nascent Z-discs before actin was observed in a striated pattern is consistent with the hypothesis that CapZ directs the location and polarity of actin filaments during I-band formation in skeletal muscle cells. The assembly of CapZ at nascent Z-disc structures also was observed relative to the assembly of sarcomeric alpha-actinin, titin, and thick filaments. Titin and myosin were observed in structures having the organization of mature sarcomeres prior to the appearance of CapZ at nascent Z-discs. The distribution of CapZ and sarcomeric alpha-actinin in young myotubes was not coincident; in older myotubes, both CapZ and alpha-actinin were co-localized at Z-discs. In cardiac myocytes, CapZ was detected at Z-discs and was distributed in a punctate pattern throughout the cytoplasm. CapZ also was co-localized with A-CAM and vinculin at cell-cell junctions formed by the myocytes. PMID- 8402954 TI - Preferential dendritic localization of pericentriolar material in hippocampal pyramidal neurons in culture. AB - Centrosomes are unique cytoplasmic structures which serve as microtubule organizing centers (MTOC). In most animal cells centrosomes consist of one or more pair of centrioles surrounded by electron dense amorphous pericentriolar material (PCM) responsible for nucleation of microtubules. In the present study we analyzed the pattern of induction and localization of proteins of the PCM at different stages of neuronal development in cell cultures prepared from the embryonic hippocampus. For this purpose we used a human polyclonal antibody that recognizes two proteins of the PCM (100 kd and 60 kd, respectively). The results indicate that in mature neurons, pericentriolar immunoreactive material is preferentially localized in dendritic processes, and that throughout the course of neurite development and differentiation it is systematically excluded from the neuron's axon. Western blot analysis showed that during neuronal development in situ, there is an increase in the immunoreactivity for both proteins recognized by this antibody. In contrast, in hippocampal pyramidal neurons that develop in culture, there is an increase in the 60 kd polypeptide, while the 100 kd one is not detected after 7 days in vitro. PMID- 8402955 TI - Microtubule behavior in PC12 neurites: variable results obtained with photobleach technology. AB - We have examined the effects of various means of photobleaching on the recovery of fluorescence, movement, and morphology of the microtubules in the neurites of rhodamine-tubulin-injected PC12 cells. We find that, depending on power of and time of exposure to the bleaching beam, we can generate at least three different patterns of fluorescence recovery in regenerating PC12 neurites. If bleaching is performed with a relatively low-power beam for an extended period, fluorescence in polymer recovers very little after 1 hour. Under these conditions, however, tubulin immunostaining is seen extending through the bleach zone, and microtubules are present through the bleached zone in thin section electron micrographs. If bleaching is performed with a high-power laser, for 0.5-5 seconds, fluorescence recovery also is quite slow, but electron microscopic observations reveal that no microtubules extend through the bleached region of the neurite, and the uranyl acetate-stained cytoplasm appears more electron lucent than in the unbleached neurite. Finally, if bleaching is performed by very brief exposure to a high-intensity laser beam, resulting in an incomplete reduction of fluorescence intensity through the bleach zone, fluorescence recovery occurs within 20-30 minutes, and immunostained microtubules appear intact through the bleach zone; electron microscopy confirms that microtubules extend through the bleached zone of such neurites. In all three cases, movement of the bleach zone is observed in approximately half of the experimental neurites. These results indicate that highly variable microtubule behaviors can be obtained with photobleach technology, presumably due to different levels and pathways of photodamage generated by different bleach protocols. Nevertheless, it is clear that both turnover and movement of microtubules occur in PC12 neurites, and both are likely to be involved in neurite maintenance and growth. PMID- 8402956 TI - Expression of four myosin heavy chain isoforms with development in mouse uterus. AB - In smooth muscle tissue, two or three isoforms of myosin heavy chain (MHC) have been reported (SM1, SM2, and/or NM). In mouse uterus tissue, four bands in the region of the MHC's can be resolved on high resolution SDS polyacrylamide gels. Western blots using smooth muscle (SM) MHC-specific and nonmuscle (NM) MHC specific polyclonal antibodies show the upper two bands in the MHC region are SM isoforms, whereas the lower two bands are NM isoforms. One-dimensional peptide maps of these four bands show each to have a unique pattern of polypeptide fragments following alpha-chymotrypsin digestion. Developmental expression of myosin heavy chains (MHC) in mouse uterus, aorta, bladder, and stomach (6 ages, 10-150 days) was determined using tissue homogenates. In the uterus, both SM MHC's show an increase in relative content with increasing age, whereas the NM MHC's show a decrease. The mouse aorta shows a significant increase in the SM MHC's and a significant decrease in the NM MHC from day 10 to day 30, which is similar to data reported for the rat aorta. Whereas both the bladder and stomach contain relatively small amounts of NM MHC's (approximately 10% or less), these quantities do show decreases with development. The SM1:SM2 ratio for the uterus remains high (3.4 at 150 days) through development; the aorta, bladder, and stomach also start out high, but tend toward 1.0 in the 150-day animals. The presence of four MHC isoforms in the uterus with unique developmental regulation of expression is consistent with hypotheses of unique functional roles for these isoforms. PMID- 8402957 TI - [Problems in receiving and dispensing in Czech pharmacies]. PMID- 8402958 TI - [75th anniversary of the State Institute for Drug Control. Its evolution, present status and perspectives]. PMID- 8402959 TI - [Herbal drugs in Kampo preparations]. AB - Medicaments of the Kampo medicine are composed from vegetable drugs. This article describes drugs unknown in European medicine and used in Kampo preparations- Radix bupleuri, Tuber pinelliae, Hoelen, Radix scutellariae, Cortex magnoliae, Fructus zizyphi, Herba perillae, Radix paeoniae, Rhizoma atractylodis, Rhizoma alismatis, Radix alismae, Rhizoma cnidii, Radix angelicae, Rhizoma coptidis, Fructus gardeniae, Cortex phellodendri, Polyporus, Radix ginseng. PMID- 8402960 TI - [Determination of heavy metals in samples of herbal drugs using differential pulse polarography]. AB - The contamination of the environment requires the content of heavy metals in medicinal plants to be limited and controlled. The present paper aims to demonstrate the possibilities of application and advantages of a modern polarographic method, differential pulse chromatography (DPP), in the identification and determination of heavy metals in the plant material. In the plant drugs Flos chamomillae and Flos calendulae, the heavy metals Pb, Cd, Ni, Zn, Cu and Fe were identified and determined. The World Health Organization (WHO) indicated Pb, Cd and Ni as dangerous toxic trace elements. PMID- 8402961 TI - [Protective effect of propentophylline (HWA 285) in ischemic spinal cord injury]. AB - The effect of propentophylline (HWA 285) on the course of the functional regeneration of the hind extremities and on the regeneration of the energetic metabolism in the spinal cord after ischaemia produced by ligating the abdominal aorta for 20 or 30 mins was investigated. Administration of 20 mg/kg i.v. propentophylline after the induction of ischaemia produced marked regeneration of these parameters in comparison with ischaemia without administration of the drug in the course of four days. The protective effect of propentophylline administered prior to ischaemia was markedly weaker. PMID- 8402962 TI - [Local anesthetics. CX. Preparation, activity and partition coefficients of pyrrolidinoethylesters of 2-, 3- and 4-alkoxy-substituted phenylcarbamic acids]. AB - By pyrrolidinoethylalcohol addition on 2-, 3- and 4-alkoxy-substituted phenylisocyanate in non-aqueous toluene medium a series of 30 compounds of the type Pyrrolidinoethylesters of 2-, 3- and 4-alkoxy substituted phenylcarmabim acids were prepared. Pharmacological evaluation results shown after comparing with cocaine and procaine standards that these compounds have a higher index of effectiveness in both types of local anaesthesia. The maximum effectiveness of surface anaesthesia shows Pyrrolidinoethylester of 2-octyloxy phenylcarbamic acid which is 204-times more effective as cocaine. The maximum effectiveness of infiltration anaesthesis shows Pyrrolidinoethylester of 2-heptyloxy phenylcarbamic acid which is 66-times more effective as procaine. Prepared compounds are more effective in the case of surface anaesthesia as in the case of infiltration anaesthesia and they have comparatively high values of partition coefficients P' and P' cal. Comparison of prepared compounds effectiveness and analogic piperidinoethylesters shows that in more cases they are either equal or nearer effective. The values of their P' and P' cal correlate with these results. PMID- 8402963 TI - [Salvia officinalis l. I. Botanic characteristics, composition, use and cultivation]. AB - Salvia officinalis L. is an essential oil containing plant, which does not wildly grow in the territories of the Czech and Slovak Republics but it can be successfully cultivated. It is a perennial half-shrub, from which non-flowering herbaceous sprouts or leaves are collected for pharmaceutical purposes. After drying at a temperature not exceeding 35 degrees C they are the plant drugs Herba salviae or Folium salviae. In PhBs, Herba salviae is official. The drug contains mainly ethereal oil (1-2%), diterpenes, triterpenes and tannin. The pharmacopoeial criterion of quality is the content of essential oil, which is produced in an increased amount in the plant in warm summer months. Herba salviae and the extracts prepared from it are used as an antiseptic agent, an antiphlogistic agent, in the inflammations of the oral cavity and gingivitis and also as a stomachic and an antihydrotic agent. Its utilization in cosmetics and food industry is also of importance. PMID- 8402964 TI - [The effect of calcium channel blockers in experimental myocardial infarct in rats]. AB - The effect of the blockers of calcium channels on the development of myocardial ischaemia in rats with an occlusion of the coronary artery was examined. An occlusion of the coronary artery was carried out in rats anaesthetized with pentobarbital by tightening the ends of the ligature freely placed under the left coronary artery - ramus interventricularis seven days prior to ligation. The ischaemia-induced changes in the R-wave and ST-segment were recorded using ECG. The occlusion of the coronary artery produced arrhythmias, a significant elevation of the ST-segment and a slight increase in the heart rate. The blockers of calcium channels with different pharmacological properties - verapamil, nifedipine and diltiazem influenced the ischaemia-induced changes with different intensity. Nifedipine (0.02 mg.kg-1, i.v., 30 min prior to occlusion), verapamil (0.2 mg.kg-1, i.v., 10 mins prior to ischaemia), and diltiazem (0.3 mg.kg-1, i.v., 10 mins prior to ischemia) significantly reduced the increased elevation of the ST-segment. The highest effect on the above-mentioned model was shown by verapamil. PMID- 8402965 TI - [Local anesthetics. VXII. Preparation and effectiveness of dibasic alkylesters of 2-, and 3-alkoxy-substituted phenylcarbamic acids]. AB - In a series of 24 compounds of the type of dibasic alkylesters of 2-, and 3 alkoxy substituted phenylcarbamic acids, esterified alcohols 1-dipropylamino-3 pyrrolidino (piperidino or perhydroazepino)-2-propanol were used. It follows from the results of pharmacological evaluation that these compounds possess a high index of effectiveness in surface and infiltration anaesthesia compared to the standards of cocaine and procaine. Maximum effectiveness in surface anaesthesia was shown by [1-dipropylaminomethyl)-2- (piperidino)ethyl]-ester of 3 butoxyphenylcarbamic acids whose activity is as much as 180 times higher than that of cocaine. Maximum effectiveness in infiltration anaesthesia was shown by [1-dipropylaminomethyl)-2-(pyrrolidino)ethyl] ester of pentyloxyphenylcarbamic acid achieving an activity 250 times higher than that of standard procaine. The prepared compounds are more effective in the case of infiltration than in the case of surface anaesthesia. PMID- 8402966 TI - [Anti-inflammatory activity of aqua-(aryloxyacetate) copper complexes]. AB - The anti-inflammatory activity of selected mononuclear Aqua(aryloxyacetato)copper(II) complexes with the composition [Cu(R-O CH2COO)2(H2O)n].mH2O (R = phenyl, n = 3; m = 0), 4-chlorophenyl (2; 0), 4-chloro 2-methylphenyl (2; 0) and naphthyl (2; 2) was examined using rat paw dextrano edema. The antiphlogistic effects of the tested complexes were compared to those for corresponding aryloxyacetic acids and salicylic acid and its Cu(II) salt. The compounds were administered i.p. in a single effective dose of 10 mg/kg. The complexes with R = phenyl, 4-chlorophenyl and 4-chloro-2-methylphenyl exhibited a biological activity comparable to that of copper(II) salicylate tetrahydrate. PMID- 8402967 TI - [Antimicrobial activity of diazole-(N-salicylidene-L-alpha- alaninato)-copper complexes]. AB - The antimicrobial activities of a group of copper(II) chelates with the composition [Cu(TSB)(L)] or [Cu(TSB)(L')]. (TSB = N-salicylidene-L-alpha alaninate dianion, L = imidazole and its alkyl derivatives, L' = pyrazole and 3,5 dimethylpyrazole) were studied. Bis-diazole complexes with L' = pyrazole and 3,5 dimethylpyrazole were found to be the most active against S. aureus (MIC = 78 and 20 micrograms.cm-3), less effective against C. albicans (156 micrograms.cm-3), T. terrestre and M. gypseum (100 micrograms.cm-3). The observed biological properties of complexes are discussed in relation to their proposed structures. PMID- 8402968 TI - [The past and the future of the Czech Pharmaceutical Society]. AB - The main features of development of the Pharmaceutical Society as the only scientific, specialist and professional organization in the years 1950-1990 are surveyed. Increasing activities of the Society, extenvicity of the arranged actions and increasing interest of pharmacists to participate are stressed. Attention is drawn to the regular changes in the activity of the Society in the present time which result from the establishment and activity of the Pharmaceutical Chamber and the need to intensity the trade union movement. The Czech Pharmaceutical Society must continue tu guarantee the scientific standard of Czech pharmacy and appropriate foreign co-operation. Pharmacists working in the individual pharmaceutical sciences should assume positions in the management of the Society and its journal. PMID- 8402969 TI - [Preparation of the first peroral antibiotic, Anginol-tablet]. AB - In 1892-1896 Ivan Honl and Jaroslav Bukovsky successfully tested pyocyaneoprotein (pyocynase), later used as the pharmaceutical preparation Tonsilan, in the therapy of crural ulcers and other bacterial diseases. In 1911 Honl prepared pyocyaneoprotein as the first antibiotic in the form of tablets called Anginol tablets. Anginol was used in the Czech Lands in infectious diseases of the pharynx and larynx, particularly in tonsillitis, until the end of the Second World War. The present paper reports the procedure of compounding Anginol-tablets and their composition. PMID- 8402970 TI - [Problems at the hospital pharmacy in Kladno]. PMID- 8402971 TI - [Phytotherapeutic aspects of diseases of the circulatory system. 2. The oyster mushroom and its potential use]. AB - The oyster fungus (Pleurotus ostreatus (Jacq. ex Fr.) Kumm., Pleurotaceae) is a wood-worming fungus with a significant use in the food industry. Out of the products of the primary and special metabolisms, which were found in fruit bodies in the 1970s, it is possible to mention mainly lipids, sterols, lipophilic vitamins, vitamins of the B group, mono-, oligo- and polysaccharides, amino acids, peptides, some enzymes and derivatives of aliphatic hydrocarbons as fragrant substances. Some of these substances exert antiviral, antineoplastic and hypocholesterolaemic activities. In the case of the hypocholesterolaemic effect, not all substances that take part in it have been isolated and identified yet. The fruit bodies of the oyster fungus became a basis for some dietetic preparations in the Czech Republic and Slovakia, e.g. HLIVETA Eritaden and PLEUROS 600. They are usable for the prevention and adjuvant treatment of hypercholesterolaemia. Besides this, the oyster fungus is a source of other substances that can be used in other fields of prevention or treatment. PMID- 8402972 TI - [Polyvalent antibiotics in ambulatory practice]. AB - The present paper analyzes the consumption of antibiotics from the viewpoint of the diagnosis, specialization of the prescribing physician, and the age and sex of the out-patients. The analysis concerns the prescription within two months in a pharmacy at a policlinic. The results are presented according to the frequency of diagnosis, polyvalence of the antibiotics and the specialization of the prescribing physician. PMID- 8402973 TI - [Immunomodulating activity of ethanol-water extracts of the roots of Echinacea gloriosa L., Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth tested on the immune system in C57BL6 inbred mice]. AB - The ethanolic extract from the roots Echinacea gloriosa L. (Moench), Echinacea angustifolia DC. and Rudbeckia speciosa Wenderoth shows immunomodulating activity. It was seen on the seventh day after five days of in vivo treatment of mice. The most marked immunostimulatory effect was observed on the lysosomal and peroxidal activity of peritoneal macrophages, and splenic cells after in vivo treatment with the ethanolic extract of the roots of R. speciosa Wenderoth. PMID- 8402974 TI - [Anti-pyretic activity of aqua-(diacetoxybenoate)-copper complexes]. AB - The antipyretic activity of binuclear (diacetoxybenzoato)-copper(II) aquacomplexes [Cu2(2,Y-DAB)4(H2O)2] (Y = 4 and 5) as well as cupric acetylsalicylate [Cu2(AcSal)4] was assayed in a rabbit model of endotoxin-induced fever. The complexes and uncomplexed carboxylic acids (administered i.p. in a dose of 20 mg/kg) exhibited even a subnormothermic course in lowering of febrile animals' body temperature in the order: 2,5-diacetoxybenzoic acid (-0.09 degree C) < 2,4-diacetoxybenzoic acid (-0.35 degree C) < Cu(AcSal)2(-0.39 degree C) approximately equal to acetylsalicylic acid (-0.40 degree C) < Cu(2,4-DAB)2.H2O ( 0.61 degree C) < Cu(2.5-DAB)2.H2O (-0.97 degree C). On the other hand, the mean value of untreated fevered animals was +0.67 degree C. PMID- 8402975 TI - [Cryopreservation of embryos in the in vitro fertilization/embryo transfer program]. AB - An ultra-rapid method for cryopreservation of human embryos, using 1,2-propane diol, was developed. 280 spare embryos obtained in the IVF program were frozen by this method. 91 embryos were thawed and 68 of them (74.7%) survived the procedure with more than half the blastomeres intact. 65 embryos were transferred in 28 cycles. Two pregnancies developed (7.1%), one of which ended by the birth of a healthy girl (the first birth after transfer of a frozen/thawed embryo in the Czech Republic). The other pregnancy is ongoing. PMID- 8402976 TI - [New approaches in the treatment of hypotrophic fetuses]. AB - Hypotrophy or intrauterine growth retardation of the foetus associated with placental insufficiency still remains a serious obstetric problem. In the submitted paper the authors compare hypotrophic foetuses treated in 1989 and 1990 at the authors' clinic by conventional methods only (biometry, bed rest, CTG oestriol, glucose and possibly heparin) with a group of hypotrophic foetuses treated in 1991 and 1992 with magnesium and where the therapeutic effect was followed up by Doppler flowmetry. Based on the results, the demand to improve placental insufficiency by pharmacological induction and stimulation of specific placental enzymes is met perfectly by concomitant parenteral and oral magnesium administration to pregnant women. PMID- 8402977 TI - [Significance of small tissue samples in the prevention of cervical carcinoma]. PMID- 8402978 TI - [Cytomorphology in the diagnosis of endometrial carcinoma and its prognosis]. PMID- 8402979 TI - [Shoulder dystocia]. PMID- 8402980 TI - [Pregnancy in women with autoimmune diseases]. PMID- 8402981 TI - [Endometriosis and osteoporosis]. PMID- 8402982 TI - [Endometriosis in monkeys and its relation to radiation exposure]. PMID- 8402983 TI - [Use of staplers in cesarean section]. PMID- 8402984 TI - [Maternal mortality in the Czech Republic 1990-1991]. PMID- 8402985 TI - [The professional concept of gynecology and obstetrics]. PMID- 8402986 TI - [Why we should adopt foreign rate schedules]. PMID- 8402987 TI - [Private insurance in Austria]. PMID- 8402988 TI - [Method of developing general principles of perinatal care]. PMID- 8402989 TI - [Lege artis and non lege artis approaches in obstetrics and gynecology]. PMID- 8402990 TI - [Perinatal and maternal mortality in Sweden]. PMID- 8402991 TI - [Insurance and legal protection of the pregnant woman and obstetrician in Sweden]. PMID- 8402992 TI - [Embolisms and postpartum hemorrhage]. PMID- 8402993 TI - [Infections during pregnancy and maternal mortality in the Czech Republic]. PMID- 8402994 TI - [Proceedings of the conference of the Czechoslovak Gynecological and Obstetrical Society. 4-6 November 1992, Karlovy Vary]. PMID- 8402995 TI - [Surgical complications and maternal mortality in the Czech Republic]. PMID- 8402996 TI - [Severe forms of late gestoses]. PMID- 8402997 TI - [Serious cardiovascular diseases in pregnancy]. PMID- 8402998 TI - [Maternal mortality 1978-1990 and its main causes]. PMID- 8402999 TI - [Prenatal diagnosis of congenital developmental defects]. PMID- 8403000 TI - [Prematurity]. PMID- 8403001 TI - [Diagnosis and therapy of fetal hypotrophy]. PMID- 8403002 TI - [Prenatal care]. PMID- 8403003 TI - [Prenatal care of serious complications and pathologic conditions in pregnancy]. PMID- 8403004 TI - [Prenatal care: ultrasonic diagnosis]. PMID- 8403005 TI - [Care during labor]. PMID- 8403006 TI - [Perinatal care from the viewpoint of the neonatologist]. PMID- 8403007 TI - [Concluding recommendations for care related to decreasing maternal mortality in the Czech Republic]. PMID- 8403008 TI - [General principles of prenatal and intrapartum care]. PMID- 8403009 TI - [The role of anesthesiology and resuscitation in high-risk pregnancy]. PMID- 8403010 TI - [Pars plana vitrectomy in epimacular membranes]. AB - Epimacular membranes /EMM/ reduce markedly visual acuity and the only treatment is pars plana vitrectomy /PPV/ and removal of the EMM. The authors analyzed the results of PPV in 30 eyes with EMM of different aetiology. The group comprised 4 idiopathic EMM and 26 secondary EMM after operations of a detached retina, injury, intraocular inflammation, occlusion of the retinal vein and retinopathies of the premature. The operation markedly improved the visual acuity in 20 eyes /67%/ and the best results were obtained in idiopathic EMM. The prognosis did not depend on the preoperative visual acuity and duration of the disease and was determined by the state of the macula beneath EMM which in dense secondary EMM is difficult to assess before operation. The poorest functional results were obtained in postinflammatory and post-traumatic EMM. PMID- 8403011 TI - [Initial experience with block excision of choroidal tumors]. AB - The authors present their experiences with the microsurgical method of therapy of choroidal malignant melanoma. During the last two years 10 patients have been admitted to the hospital that had malignant melanoma of uvea, eight of them had choroidal melanoma. In 5 patients local excision of choroidal melanoma was made, according to Peyman's criteria. One patient has reached the visual acuity of 5/10, other two patients had vitreous hemorrhages after the operation without absorption. Fourth patient has developed hypotony and in one patient the operation had to be interrupted and the enucleation of the eye was made. Local excision seems to be a promising operation method, which can preserve the eye and its good visual acuity in indicated cases. PMID- 8403012 TI - [Initial experience with implantation of disc-type intraocular lenses]. AB - The authors present their postoperative results of extracapsular cataract extraction with implanted disc intraocular lens CD 801 A (Pharmacia). In 1991 within 6 months 62 of these lenses had been implanted. A year after the operation, the location of the intraocular lens, transparency of posterior capsule, the shape of the pupil and visual acuity (correction) had been followed. In 6 patients (9.7%) dislocated lenses had been found, 11 patients (17.7%) developed decreased transparency of the posterior capsule, and in 30 patients (48.4%) circular shape of the pupil was in different way distorted. 48 eyes (77.42%) had 1 dioptre error or less, 58 eyes (93.55%) had 2 dioptres error or less and 4 eyes (6.45%) had an error of more than 2 dioptres. PMID- 8403013 TI - [Implantation of intraocular lenses in diabetes mellitus--experience from 1989 to 1991]. AB - The authors report favourable results with implantation of an intraocular lens after extracapsular extraction in diabetic subjects. In 1989 they implanted only one intraocular lens in 49 eyes without diabetic retinopathy, in 1991 in 110 eyes from 111 without diabetic retinopathy and 12 eyes from 26 with diabetic retinopathy. The incidence of complications did not differ significantly from that in non-diabetic patients. In 76.6% eyes without diabetic retinopathy 6 months following implantation of an intraocular lens the vision was 6/12 or better, in eyes with diabetic retinopathy this applied to 62% of the eyes. In the course of 6 months following operation diabetic retinopathy developed in 5 of 110 eyes. To achieve long-term favourable results it is necessary to check the patients regularly after operation and in case of progressing diabetic retinopathy prompt laser photocoagulation is indicated. PMID- 8403014 TI - [Development of visual acuity after sutureless scleral tunnel cataract surgery]. AB - In 164 patients with cataract/208 eyes/operated by endophacoemulsification using scleral tunnel without suture, the postoperative development of corrected visual acuity was investigated. The best results were recorded after operation by scleral tunnel 5 x 5 mm in size. PMID- 8403015 TI - [Manual phacofragmentation in extracapsular cataract extraction]. AB - The author describes three different techniques of manual phacofragmentation in extracapsular extraction of cataract. Based on the experience with the first 60 operations made by these techniques, the author recommends some principles which should be respected. In the mentioned group only once the posterior capsule was damaged in conjunction with division of the nucleus. When adequate care is not taken or when a poor techniques is used, k. striata occurs more frequently but disappears within one week. The author considers the described technique as a possible and suitable alternative of classical extracapsular extraction of cataract in departments where FAKO is not available. PMID- 8403016 TI - [Suture technic in perforating keratoplasty and postoperative astigmatism]. AB - The authors evaluate postoperative vision, value of astigmatism and keratometry in 25 patients operated in 1992 by perforating keratoplasty, incl. 11 where the disc was fixed by interrupted stitches and 14 by continuous diagonal stitches. After evaluation of the influence of interrupted stitches and continuous diagonal stitches on postoperative astigmatism in perforating keratoplasty the authors recommend the use of diagonal suture. The reason is in the first place the uniform spreading and good adaptation of the transplanted cornea, as well as slighter traumatization and easier removal of stitches. An important role is played also by the larger number of measurable radii of the corneal curvature with regard to possible postoperative correction of astigmatism either by added stitches or differentiated adjustment of the tension of the continuous stitch. PMID- 8403017 TI - [HLA tying in endogenous uveitis]. AB - In a group 67 patients with endogenous uveitis without systemic disease the authors typed, using the microcytotoxic test, HLA antigens class I. The results were processed by statistical methods and compared by the chi 2 test and Fisher's exact test of probability with a control group of the healthy Plzen population. The following genes were classified as risk genes: A 30, B 5, B 27 and Cw 2. PMID- 8403018 TI - [Tortuous retinal vessels]. AB - The authors observed an occurrence of tortuosity of retinal vessels from the group of 5,000 investigations of an ocular fundus. Elimination of all local reasons or symptoms of general diseases, even in spite of the great possibility of modification of this feature, three characteristic types were found. In two of them the familiar and hereditary occurrence was proved. PMID- 8403019 TI - [The attitude of patients to glaucoma]. AB - The authors are concerned since 1987 with a relatively large group of patients suffering from glaucoma, whereby attention is focused on social and psychological factors in conjunction with glaucoma. Examinations are made, using a binary questionnaire. In addition to already known facts on the marked influence on work capacity and problems ensuing from the necessity of a relatively frequent need to administer eye drops the authors found that glaucoma has a great impact on the patients' life. It has an impact on family life, in some instances the disease caused loss of the partner. 60% of the patients frequently think of their disease frequently and some 8% of the patients feel that life is not worth living. 6% of the patients in the group contemplated possible suicide. The authors feel that the group at greatest risk are old women and men who as a result of the disease are complete or partial invalids. PMID- 8403020 TI - [How do animals see?]. PMID- 8403021 TI - [Ultraviolet rays and the eye]. PMID- 8403022 TI - [A course on the fundamentals of psychotherapy for general physicians]. PMID- 8403023 TI - [Jaromir Mundi, Doctor of Medicine and Surgery, and his role in the humanization of psychiatry in the last century]. PMID- 8403024 TI - [Bulimia nervosa--therapeutic possibilities with emphasis on cognitive behavioral therapy]. AB - The author pays attention to recent trends in the treatment of bulimia nervosa, he discusses problems of therapeutic methods and hospital treatment. Special attention is paid to ambulatory and cognitive behavioural therapy. The author emphasizes its integrating character due to the complexity of the problem of eating disorders. The latter are becoming one of the most serious health and social problems of adolescent women not only in western Europe and the USA but also in this country. The author defines some principles of the cognitive behavioural approach in ambulatory treatment of bulimia which is briefly described in the conclusion of the article. He reminds that it is always essential to take into account the grade of the disorder, the differential character of its psychosocial picture and the patient's needs. This should be matched by the offered therapeutic methods using different possibilities of in- and out-patient treatment. An essential prerequisite of treatment is not only detailed knowledge of the therapeutic method but also of problems associated with impaired food intake. PMID- 8403025 TI - [Review of findings on binding of tricyclic antidepressants]. AB - Authors summarize knowledge in the field of the high-affinity binding of tricyclic antidepressants on cell membranes within last 12 years. Some ideas about the significance of such binding are stated. Both the data from papers and our knowledge are discussed with regard to the possibility of their clinical application at depressed patients. Effects of the tricyclic antidepressants are linked with their influence on the active transport of neuromediators through the cell membrane but their molecular mechanisms are unknown. Present-day state of research is described and the orientation on the role of membrane phospholipids in the binding studies of the tricyclic antidepressants is emphasized. PMID- 8403026 TI - [Diagnosis of mixed psychoses]. AB - The author presents a review of views on so-called "medium or mixed" endogenous psychoses, starting with Kraepelin and Bleuler to the present time, the creation of the term "schizoaffective psychosis (disorder)" in diagnostic classifications. He draws attention to the still persisting influence of Kraepelin-Bleuler diagnostics, incl. their controversial character as regards the classification of so-called "mixed psychoses". Other diagnostic concepts were either completely or are enforced with great difficulties. On the diagnosis of schizoactive psychosis the author demonstrates the changing understanding of its contents and thus also the controversial attitude to the diagnosis in the course of time which was reflected in the development of diagnostic classifications. The paper also comprises the author's study of the frequency of diagnosis of endogenous psychic disorders in the Psychiatric Research Institute (present name Psychiatric Centre Prague) in the course of 29 years. The study indicates that from the sixties the diagnosis of schizoaffective disorder played the role of a buffer between the schizophrenic and manic-depressive area. It helped to solve ranae of the unstable concept of schizophrenia and classification of atypical forms of endogenous psychoses. The study indicates that there exists possibly a closer relation between the schizoaffective disorder and affective disorders than with schizophrenia. In the conclusion the author appreciates the asset of classifications DSM-III and ICD-10 for the definition and differentiation of schizoaffective disorders. PMID- 8403027 TI - [Ritanserin in autistic children]. AB - Ritanserine had a positive effect on children with Rett's syndrome and children with autistic syndrome and mental retardation. The most marked effect was observed in reduction of psychomotor instability, impaired concentration of attention and partly also in autism. PMID- 8403028 TI - [EMG findings in patients with diabetic polyneuropathy treated with sabeluzole]. AB - Author trie to prove a possible positive effect of sabeluzole in patients with diabetic polyneuropathy. All patients were examined by EMG stimulating method for n. tibialis sin., n. peroneus sin. and n. medianus dx. The distal motor latency, motor conduction velocity and sensory conduction velocity with areas of motor evoked responses were evaluated. The realised test had lower sensitivity than the originally planned one and the expected positive effect of sabeluzole was not proved by this method. PMID- 8403029 TI - [Sabeluzole and flunarizine in memory processes: animal studies]. AB - The memory enhancing properties of sabeluzole nad flunarizine were evaluated in two experimental paradigms in male rats. First, we studied the protective action of both drugs against a chlordiazepoxide-induced impairment of habituation. Sabeluzole (5 or 25 mg/kg) or flunarizine (10 mg/kg) were administered sc 1 hr before and chlordiazepoxide (20 mg/kg) immediately after the acquisition session. In the retention session 72 hr later, chlordiazepoxide treated animals displayed higher locomotor and exploratory activities and this effect was blocked by pretreatment with sabeluzole or flunarizine. The results suggest that both drugs prevented amnesic effect of chlordiazepoxide. The second paradigm was the social memory test in which the time spent in social investigation behaviour toward a familiar or a novel juvenile conspecific was used to measure the duration of the memory that the animal forms of the juvenile. Sabeluzole (25 mg/kg) or flunarizine (3 mg/kg) were injected to the adults immediately after the initial exposure. Reexposure to the same or a novel juvenile was done 2 hr later. In contrast to controls, sabeluzole-treated animals showed a significant reduction in social investigation during reexposure to the same juvenile. Since there was no effect on investigation of a novel juvenile, results suggest that sabeluzole treated rats are able to remember longer the individual characteristics of juvenile rats obtained through olfactory cues during a short social interaction. The time spent by adults treated with flunarizine in the investigation the same juvenile during reexposure was similar to that observed during the initial exposure. It means that flunarizine was ineffective in enhancing short-term olfactory memory. PMID- 8403030 TI - [3 years of the "new house" at the Psychiatric Clinic of the Medical School Hospital in Olomouc]. AB - The author present an account on changes in the operation of the psychiatric clinic made possible by acquisition of a new building for the Clinic in 1989. This made it possible to improve conditions for patients, staff and teaching. The author discusses experience assembled during the first three years of operation of the "New House". PMID- 8403031 TI - [A comparative biology and dynamic interpretation of necrophilia]. AB - The authors discuss some aspects of Necrophilia based on classic Freudian instinctive theories. Parallels are seen between different levels of life development on Earth and the basic antagonistic drives (Eros and Thanatos) in an attempt to explain this rare and unusual paraphilia. The authors mention, that in unicellular organisms, e.g. Amebas and Schizomycetae, there often does not exist what could be called "the death of an individual", when by splitting two new "individual organisms" are created and no death as such occurs. Here the supposed antagonism of these basic drives-Eros and Thanatos-actually does not manitest itself in the concentional sense. Necrophilia could be interpreted as a regressive desire to return into a phylogenetically older stage of life development, where no individual dies, and life continues without interruption. (On the level of these unicellular organisms, we should speak about "Dividuum" instead of using the classic term "Individuum".) There is some analogy to this regressive desire at a higher level of the biological development in Human society--a magic conviction about the possibility to receive a dead person-common in preliterate cultures. Necrophilia represents an attempt at symbolic unification between antagonistic active and passive drive tendencies and between the Libido and the Destructive instinct. PMID- 8403032 TI - [Sexual stimulation and the female orgasm]. AB - Questionnaire data regarding mode of sexual stimulation were collected in 200 women treated for neurotic disorders and 100 female health professionals and counselors. Clitoral stimulation was the source of female orgasm in 90% of subjects, while three quarters achieved orgasm also by means of vaginal stimulation. The anterior wall of the vagina seemed to be slightly more important than the posterior one. One third of the respondents reported effective stimulation in the depth of the vagina with cervical tapping. Stimulation in the area corresponding to the alleged G spot was acknowledged as effective by 10 to 20%. The sexual responsiveness of neurotic patients was generally somewhat lower than responsiveness of health professionals and counselors. PMID- 8403033 TI - [Sexual asphyxiophilia (Koczwarism)]. AB - The author discusses the definition, history, anthropological data and sexual differences of a deviant sexual activity, asphyxiophilia (Kocwarism), the objective of which is stimulation or enhancement of sexual excitation or induction of orgasm by induced hypoxia. The author draws attention to the basic features which provide evidence of this life threatening and so far inadequately diagnosed sexual practice in both sexes, in particular in women. These features can be used in forensic psychiatry and sexuology and in forensic medicine. PMID- 8403034 TI - [Epilepsy and intelligence. II. Correlation among 13 variables]. AB - In a group of 215 ambulatory adult patients the authors assessed links with full scale IQ, verbal IQ, performance IQ and different Wechsler-Bellevue sub-tests. They revealed marked correlations in particular with the frequency of grand mal seizures, invalidity and the onset of the disease. More important than the close correlation of IQ and education is the finding that an earlier onset of the disease markedly reduces the opportunities of good education. Less marked but still significant are correlations with the known aetiology of the disease, the duration of the disease, the frequency of other seizures than grand mal and the course of the disease. Dramatically increased correlations with the frequency of other seizures than grand mal were recorded in a sub-group of 77 men whose data on the disease can be considered valid (in WPSI, the Washington Psychosocial Seizure Inventory, values of scale L within the range of 0-3). No marked correlations were found with seizures in relatives. The position of bachelors is often adverse also as far as intellectual performance is concerned. Patients with a history of contact with psychiatry have lower intellectual performance. Patients who admit subjective needs of psychiatric or psychological assistance have a markedly better performance. It is probable that rehabilitation procedures which improve the collaboration and adherence to the therapeutic regime, in particular as regards taking prescribed drugs may promote also better cognitive performance. PMID- 8403035 TI - [Hyperlipoproteinemia in mentally retarded children]. AB - Screening revealed a significantly higher incidence of hyperlipoproteinaemia in mentally retarded children in an Institute for Social Welfare than in the healthy child population (P 1%). In the aetiology participate most markedly secondary influences ensuing in particular from complications of the basic diagnosis of oligophrenia. It is mainly a question of the action of antiepileptics, neuroleptics, the influence of viral hepatic infections and other endogenous and exogenous factors. Early diagnosis of hyperlipoproteinaemia is possible so far only by biochemical methods. It is essential for genetic counselling, for prevention of deterioration as regards mental retardation, restriction of pharmacotherapy and later for a reduced risk of development of cardiovascular disease. PMID- 8403036 TI - [The nocturnal eating syndrome (2 case reports and polysomnography)]. AB - The nocturnal eating syndrome (NES) is a nocturnal sleep disorder caused by repeated awakening and the inability to fall a sleep again unless the patient ingests some food or drinks something. In children the NES is frequent, in adults rather rare and may be associated with various pathological conditions. The latter include in particular somnabulism, periodic movements of the lower extremities, narcolepsy, chronic triazolam intoxication, probably anorexia nervosa, and other eating disorders. The authors submit two case-histories of middle aged women (30 and 37 years old) without disturbances of the day-time eating behaviour, without obvious psychopathology where the symptomatology of NES developed slowly from the age of 14 and 25 years resp. Both patients ate at night small amounts of easily consumed foods which they went to fetch in the kitchen as often as five times per night. Their behaviour was calm and aimed. Usually they did not remember the nocturnal eating. Polysomnographic examination in both patients revealed poor sleep with frequent changes of the sleep stages, with frequent awakenings, in both patients incl. even awakening from deep NREM sleep. The authors conclude that in these two patients NES with somnabulism is involved. PMID- 8403037 TI - [The suicide rate in Czechoslovakia 1975-1990]. AB - The author investigates the development of the suicide rate in Czechoslovakia in 1975-1990. From comparison of statistics in the Annual Statistical Report and statistics of UZIS he concludes that the Annual Statistical Report provides more accurate data. According to data in the latter the annual suicide rate in recent years is by 500-800 suicides higher than according to UZIS. Using the method of the smallest squares, linear trends of the suicide rate are calculated according to both sources for the Czechoslovak Republic and the Czech and Slovak Republic separately. The decline is more marked in the Czech and Slovak Republic where the suicide rate is higher and thus a convergent trend was recorded. These conclusions are supported by linear trends developmental trends obtained from the above sources differ as to the degree of convergence; while according to data in the Annual Statistical Report data in the Czech and Slovak Republic will be equal during future decades, according to data from UZIS equal levels were reached already between 1989 and 1990. The author discusses the reliability of statistics of the suicide rate, possible escapes associated with different types of collection of data and the comparability of statistics where it is obvious that they are not quite complete. PMID- 8403038 TI - [Observations of participants in a bilateral symposium of the American Psychiatric Association and the Czechoslovak Medical Society]. PMID- 8403039 TI - [Report on the Who meeting on problems of prevention and control of stress]. AB - The report informs of the Consultation organized by the World Health Organization, Regional Office for Europe in Prague, 24.-26. June 1992. It was attended by 27 experts from 15 countries. The scope of the Consultation was to review effective approaches to stress management in the community setting. Considering the fact the growing number of persons and population groups in Europe Region are exposed to major stressful situations there is great need for help either from the health and social welfare system as well as from various non professional organization. During the Consultation the definition of the stress management and prevention, examples of intervention programmes and particularly set of recommendation to the WHO and its Member States in European Region were elaborated. PMID- 8403040 TI - [The "cognitive-behavioral program" in the treatment of sexual offenders]. PMID- 8403041 TI - [The Czech Pediatric Society 1990-1993]. PMID- 8403042 TI - [Sonographic image of the most frequent abdominal and retroperitoneal malignant tumors in childhood]. AB - The author presents sonomorphological characteristics of retroperitoneal neuroblastoma, nephroblastoma, non-Hodgkin lymphoma and M. Hodgkin abdominis, hepatoblastoma and hepatocellular carcinoma at the time when the diagnosis was established. The objective of the investigation was to increase the sensitivity and specificity of ultrasonic diagnosis of the mentioned malignant tumours of the abdomen and retroperitoneum during the first sonographic examination. PMID- 8403043 TI - [The short-bowel syndrome]. AB - This study was designed to asses the clinical course of five children with a short bowel syndrome after massive intestinal resection during neonatal period. On the basis of their experience the authors analyze some properties that may be advantageous in this syndrome and discuss the most important factors, including enteral feeding at an early postresection stage, which influence a full development of the small bowel adaptive mechanisms. They point out that so called "short gut colitis", sometimes occurring during advancement of enteral nutrition, can be successfully treated by sulphasalazine. The authors come to the conclusion that the prognosis for newborn infants undergoing an excessive bowel resection is far well thanks to enhancement of the intestinal adaptation by a combination of a long-term parenteral nutrition with enteral feeding. PMID- 8403044 TI - [Present possibilities of improving the diagnosis of glomerular erythrocyturia]. AB - Examination of glomerular erythrocyturia which proved useful in the differential diagnosis of haematuria faces in practice two serious problems--the necessity to examine fresh urine and differences as regards the interpretation of criteria of glomerular erythrocyturia (from 10 to 80%). The authors examined therefore the morphology of erythrocytes in urine of 65 children with different causes of haematuria, incl. 35 patients with different forms of glomerulonephritis. They found that adding 25 mg thimerasol to 10 ml urine preserves urinary elements, incl. morphologically altered red cells for a period of 72 hours or longer. This makes it possible to examine the red cells under a phase microscope after dealing with ambulatory patients. Among various morphological forms of red cells in urine of 31 patients with glomerulonephritis in urine acanthocytes were found in a concentration above 5%, and in the majority above 20%. Acanthocyturia above 5% was not present only in 4 patients with glomerulonephritis, whereby all four suffered from mild erythrocyturia. The authors consider therefore acanthocyturia above 5% as the most reliable evidence of glomerulonephritis, in case of erythrocyturia above 10 Ery in the visual field. The criteria of acanthocyturia in children are thus lower than in adults. PMID- 8403045 TI - [Affective respiratory and reflex paroxysms--evaluation of anamnestic data, clinical manifestations and therapy]. AB - The authors elaborated data, using the retrospective method, of a group of 146 patients with affective respiratory and reflex paroxysms. They focused attention on clinical manifestations of the disease, anamnestic data suggesting possible damage or immaturity of stem structures, the influence of heredity and the family environment. They found that in 63.7% the disease was manifested before the age of 1 year, most frequently at the age of 9-12 months. The cyanotic type of paroxysms was found in 67.5% of the patients, the pallid type in 21% and 3.5% of the patients suffered from both types of paroxysms. In 27.4% perinatal risks were recorded. The influence of a family-history was statistically significant in relation to the patient's age during the first attack: in patients with a positive family-history the mean age being by 1.8 months lower. 82.9% of the patients had a normal neurological finding, the EEG was evaluated as normal in 89.6% of 125 thus examined children. Psychological examinations made in 12 children revealed in all instances anomalies of personality with a predominance of lack of compliance and adaptability to the environment. Deterioration of the health status as a result of affective respiratory and reflex paroxysms was not recorded in any of the patients. PMID- 8403046 TI - [Atrial natriuretic factor in congenital heart defects in infants]. AB - The authors examined the level of the atrio-natriuretic factor (ANF) in 7 infants with inborn heart disease at the age of 3 weeks to 5.5 months. As controls they examined 9 convalescent infants aged 1-10 months. Congenital heart disease was confirmed by clinical methods (ECG and X-ray) and echocardiographically (one dimensional, two-dimensional and pulsed Doppler echocardiography, 5MHz probe, Kontron Co.). Although a small group is involved, elevated ANF values were recorded in infants with inborn heart disease, as compared with controls. PMID- 8403047 TI - [Poisoning with a high dose of amitriptyline in a 9-year-old girl]. AB - In the submitted case-history the authors publish the case of successful treatment of a 9-year-old girl whose life was at risk after intoxication with a large dose of amitriptyline. Concurrently they deal with the social background of emotional disorders and some peculiar features in this girl with suicidal behaviour; attention is also drawn to specific features of the psychopathological picture of psychiatric child morbidity. PMID- 8403048 TI - [Thanatophoric nanism and histopathologic bone changes]. AB - The authors describe in a neonate with thanatophoric nanism histopathological changes of the bones. According to X-ray findings type I was involved, the histopathological changes corresponded to type II according to Langer's classification. The consanguinity of the parents suggested the autosomal recessive type of heredity and the presence of pseudo-diastrophic dysplasia in two grade 1 cousins is considered by the authors as an incidental coincidence. The described case confirms the assumed clinical and genetic heterogeneity of thanatophoric nanism. PMID- 8403049 TI - [School phobias in childhood and adolescence]. PMID- 8403050 TI - [Practical comments on the diagnosis of so-called functional proteinuria and suggestions for a diagnostic algorithm]. AB - The submitted paper, based on the authors' experience, summarizes basic findings pertaining to isolated proteinuria. In addition to a simple classification the authors outline in a diagram the differential diagnostic procedure in patients with isolated proteinuria. PMID- 8403051 TI - [Assistance Respiratoire Extra-Corporelle (AREC)--a simple French modification of ECMO?]. PMID- 8403052 TI - [On the article by Dr. Janda on the nephrotic syndrome]. PMID- 8403053 TI - [Changes of adrenoceptors and glucocorticoid receptor in the lungs of rats with experimental respiratory distress syndrome]. AB - With the radioligand binding assay, the maximal binding capacity (Bmax) and affinity (Kd) of alpha, beta-adrenoceptors(alpha AR,beta AR) in the lung membrane and glucocorticoid receptor (GCR) in the lung cytoplasma of rats with experimental respiratory distress syndrome (RDS) induced by oleic acid have been measured. The results demonstrated that the content of alpha AR in rat lungs increased continuously during the experiment, the Bmax at 1st, 4th and 6th hour after oleic acid injection were 139 +/- 40, 127 +/- 12, 116 +/- 25 fmol/mg protein, significantly higher than normal value (83 +/- 7, n = 8-10, P < 0.01). Meanwhile, the content of beta AR and GCR decreased continuously, the Bmax at the same time were 364 +/- 18, 307 +/- 55, 240 +/- 66 and 146 +/- 28, 153 +/- 37, 150 +/- 32 fmol/mg protein respectively, significantly lower than their normal value (490 +/- 61, 227 +/- 14 fmol/mg protein, n = 6-10, P < 0.01). The results indicate that the changes of these receptors may be of significance in the pathogenesis of ARDS. PMID- 8403054 TI - [The investigation of the correlations between left ventricular end diastolic pressure and pulmonary function parameters]. AB - Left ventricular end diastolic pressure (LVEDP) in 24 patients with coronary heart, hypertension or rheumatic heart disease (mainly aortic valve pathologic change, exclude mitral stenosis), who had unobvious clinical heart failure. Before left heart catheterization pulmonary function were tested by plethysmogram, results revealed: when LVEDP > 15 mmHg in the patients, pulmonary function parameter expectancy value percentage including FEF25-75%, V25, V50, V75, FVC, VC, FEV1.0 were obviously decreased. RV/TLC expectancy value percentage was obviously increased. Pulmonary function parameter expectancy value percentage including FEF25-75%, V25, V50, V75, FEV1.0, FVC, etc. had significant negative correlation with LVEDP (r = -0.715, P < 0.001; r = -0.699, P < 0.001; r = -0.678, P < 0.001; r = -0.671, P < 0.001; r = -0.648, P < 0.001; r = -0.516, P < 0.01; respectively). RV/TLC expectancy value percentage had significant positive correlation with LVEDP (r = 0.515, P < 0.05). The results indicate that testing pulmonary function parameter helps to evaluate left ventricular function and diagnose early (mesenchymal) pulmonary edema. PMID- 8403055 TI - [The comparison of left and right heart function with lung function in patients with rheumatic valvar heart disease]. AB - According to the characteristics of changes of heart and lung function at different stages 25 patients with rheumatic valvar heart disease could be classified into three stages: 1. The stage of normal lung ventilation and volumes; 2. Stage of obstructive ventilatory function; 3. The stage of mixed ventilatory defects associated with right heart failure. As pulmonary artery pressure increased right ventricle work became elevated and left ventricle work decreased. There was an inverse correlation of vital capacity with per minute work of right ventricle. PMID- 8403056 TI - [Renal transplantation with pulmonary infection]. AB - Ten cases of renal transplantations with pulmonary infection are reported. Nine cases were diagnosed by autopsy, one case clinically. Because of compromised immunological function, in these cases the causative microorganism, clinical manifestations, courses of the disease, therapies and result of the disease were different from those of ordinary pneumonia. The causative microorganisms were mainly opportunistic bacteria including virus, protozoa and fungus. During infection, symptoms were insidious and the disease disseminated rapidly, the mortality rate was high. Therefore in any case when the cause of the fever is unknown and no abnormal signs in the chest is find, it is necessary to take chest X-ray film immediately and repeatedly. Once diagnosis of pulmonary infection is established, series of bronchoscopic examination and lung biopsy are needed to find out the causative organism in order to treat the disease specifically, then the life of the patient can be saved. PMID- 8403057 TI - [Evaluation of detection of M. tuberculosis in clinical specimens of tuberculosis by DNA amplification]. AB - The sensitivity of detection of M. tuberculosis genomic DNA were 1pg or 10-100 bacterial cell by PCR. Only M. tuberculosis, M. bovis and BCG were positive with 165 b.p band, but all other 14 mycobacterium and 10 bacteria of non-mycobacterial tested, were negative. Of 75 sputum specimens of pulmonary tuberculosis, the positive rate of PCR were 53.3%, culture method showed only 21.3%, fast-acid staining were 25.3%. 17 non-tuberculosis lung disease were negative in three methods. Of 58 tuberculosis meningitis, the positive rate of PCR, the fast-acid staining and culture in cerebrospinal fluid were 51.7%, 8.6%, 1.7% respectively. 30 non-tuberculosis meningitis were negative in three methods. The results showed that DNA amplification is a superior method with high degree of sensitivity and specificity for rapid diagnosis of pulmonary tuberculosis and tuberculosis meningitis. PMID- 8403058 TI - [Bioassay of Lyso-PAF platelet activating factor and it's role in bronchial asthma]. AB - Plasma and lung homogenate lyso-PAF were measured by using the method of bioassay in guinea pigs with allergic asthma and asthmatic patients. The results showed that Lyso-PAF levels were increased significantly in asthmatics, and it was positively correlated with TXB2 levels. It is suggested that PAF plays an important role in bronchial asthma, which may be correlated with TXA2. The method of bioassay of Lyso-PAF is stable, reliable and special instruments are not needed. PMID- 8403059 TI - [The investigation on 100 bronchial asthma and asthmatic bronchitis cases treated with high dose beclomethasone dipropionate aerosol]. AB - 100 cases, of which sixty four bronchial asthma patients and forty two asthmatic bronchitis patients sufficiently severe to be treated with inhaled corticosteroids, were investigated. All were treated with high dose beclomethasone dipropionate (BDP) inhaler aerosol (1500-2000 micrograms/day) for three months. The results showed the total effective rate was 98.4% in bronchial asthma patients and 62.5% in asthmatic bronchitis patients. The pulmonary functions and bronchial hyperresponsiveness of most bronchial asthmatic patients were improved significantly after 1-2 weeks treatment. It is recommendable to treat the oral cortisone-depending patients with BDP inhaler. After 3 months treatment, the adrenocortical secretive ability and reserve power of the patients could be slightly damaged. The investigation also showed 8 percent subjects complained of dysphonia and 25 percent suffered from oropharyngeal candidiasis. Finally, the asthmatic relapse rate was high after cessation of the therapy. PMID- 8403060 TI - [Prognostic factors in lung cancer based on Shanghai population]. AB - Life table, Cox model and Cox multivariate model were used to measure the prognostic factors in 1417 new prevalent lung cancer cases in Shanghai population of urban area. There were 3 main prognostic factors in lung cancer as follows: a. histologic type was correlated to prognosis, among which, squamous type was the best, SCLC the worst, the factor influenced to prognosis of various types was stage and therapy. b. the earlier stage the better prognosis, and related to a optimal therapy. c. Combined therapy was good for prognosis, it was optimal for either surgical or nonsurgical therapy. Early detection, extension of combined therapy and make public of knowledge on lung cancer were encouraged. PMID- 8403061 TI - [The activities of 3 enzymes in serum and cerebrospinal fluid for diagnosis of tuberculous meningitis]. AB - The adenosine deaminase (ADA), lysozyme (LZM) and lactate dehydrogenase (LDH) of serum and cerebrospinal fluid (CSF) were determined in 36 patients with tuberculous meningitis (TBM), 47 patients with non-tuberculous meningitis (N-T BM) and 20 patients with non-central nervous system diseases(control group). The results showed that the assessment of serum and CSF ADA and LZM activity may be helpful to the differential diagnosis between TBM and N-TBM. PMID- 8403062 TI - [Nd-YAG laser treatment for tracheobronchial benign lesions]. AB - In recent 5 years, 33 cases of tracheobronchial benign lesions have been treated with the method of Nd-YAG laser via bronchoscope. The follow-up survey has shown that the curative ratio reached 66.7 percent and total effective rate came up to 78.8 percent. There was significant relationship between the therapeutic effectiveness and types of lesions (P < 0.01). It had higher cure rate in benign tumour, tuberculous granuloma and foreign body's granuloma than that of inflammatory granuloma and amyloidosis. This method is a new and effective one for treating benign pathologic changes in air passage. The indication and prevention of complications have been discussed. PMID- 8403063 TI - [The advances in the pathogenesis and treatment of acute lung injuries]. PMID- 8403064 TI - [The current status of the research on platelet activating factor and pulmonary hypertension]. PMID- 8403065 TI - [Protective effect of endothelin-antiserum on oleic acid-induced respiratory distress syndrome (RDS)]. AB - On the rat RDS model produced by intravenous injection of oleic acid, , it was for the first time found that endothelin (ET) level in bronchoalveolar lavage fluid was elevated by 3-fold: and that preadministration of ET-antiserum to rat with RDS significantly improved its hypoxemia, pulmonary edema and histologic injury; inhibit the leakage of protein and intracellular enzymes from alveoli. The results suggest that ET might play an important role in the pathogenesis of RDS, and that treatment against ET would be a new approach for RDS. PMID- 8403066 TI - Annonciade Nicolai (1933-1993). PMID- 8403067 TI - In vitro slow fluctuation in horseradish peroxidase activity. AB - In vitro slow fluctuations in the level of horseradish peroxidase activity were observed in long-range experiments (72-144 h). Besides random fluctuations, regular slow oscillatory patterns with period lengths ranging from 10.0 to 39.0 h were detected by statistical analysis. The possibility that these oscillations in enzyme activity could have reflected changes in the physical environment of the experimental setup has been thoroughly examined and ruled out. Periodic exposition of the enzyme solution, otherwise kept in darkness, to blue light illumination was shown to influence the period of the oscillations. The changes in enzyme activity were correlated with a modification of the Michaelis constant estimated using guaiacol as substrate. This result was confirmed by the action of chemical modifiers of the enzyme, such as ferulic acid and rutin. It is thought that the observed oscillations in horseradish peroxidase activity are due to spontaneous and specific changes in the tridimensional structure of the enzyme in the thermic reservoir. PMID- 8403068 TI - Circadian neuroendocrine role in age-related changes in body fat stores and insulin sensitivity of the male Sprague-Dawley rat. AB - A role for circadian neuroendocrine rhythms in the age-related development of obesity and insulin resistance was investigated in the male Sprague-Dawley rat. The phases and amplitudes of the plasma rhythms of several metabolic hormones (i.e., corticosterone, prolactin, insulin, and triiodothyronine) differed in lean, insulin-sensitive (3-week-old rats), insulin-resistant (8-week-old rats) and obese, insulin-resistant (44-week-old rats) animals. Simulation of the daily rhythms of endogenous corticosterone and prolactin by daily injections of the hormones at times corresponding to the peak levels found in 3-week-old rats reversed age-related increases in insulin resistance and body fat in older (5-6 month-old) rats. Ten such daily injections of corticosterone and prolactin in 12 14-week-old rats produced long-term reductions in body fat stores (30%), plasma insulin concentration (40%), and insulin resistance (60%) (determined by a glucose tolerance test) measured 11-14 weeks after the treatment. Alterations in circadian neuroendocrine rhythms may account for age-related changes in carbohydrate and lipid metabolism in the male Sprague-Dawley rat, and resetting of these rhythms by appropriately timed daily injections of corticosterone and prolactin may help maintain metabolism characteristic of younger animals. PMID- 8403069 TI - Circadian variation in amikacin clearance and its effects on efficacy and toxicity in mice with and without immunosuppression. AB - A once-daily dosage regimen has been recently recommended in the use of aminoglycoside antibiotics since they induce a postantibiotic effect. In choosing this regimen, one must determine the most appropriate time of day for administration of the drug. We investigated the effects of the timing of amikacin (AMK) administration on the kinetics, the efficacy against intraperitoneal infection with Pseudomonas aeruginosa, and the toxicity of AMK in mice with and without immunosuppression. We found circadian variations in the kinetics, efficacy, and toxicity of the drug in mice. Male and female ICR mice, which were housed under a light-dark (12:12 h) cycle with free food and water intake, were injected subcutaneously with AMK sulfate 50 mg/kg body wt. There was a circadian variation in AMK clearance for both sexes with the maximum value in the dark phase and the minimum in the light phase after a single administration. When AMK 500 mg/kg/day was repeatedly administered once daily for 30 days, higher toxicity was demonstrated in mice injected with the drug at the time of day with lower AMK clearance, although no difference was demonstrated in the toxicity between the two time points with different AMK clearance when AMK 1,500 mg/kg was administered in a single dose. The ED50 of AMK to cure the infected mice in the midlight phase (13:00 h) with lower clearance was significantly lower than that in the middark phase (01:00 h) with higher clearance. In contrast, the ED50 in the early light phase (09:00 h) was significantly lower than that in the early dark phase (21:00 h), although AMK clearance was no different between these two different time points. In mice premedicated with cyclophosphamide to suppress immune functions, the difference in the ED50 of AMK was still demonstrated between 13:00 and 01:00 h, but not between 09:00 and 21:00 h. The present study shows not only that there were circadian variations in both AMK clearance and toxicity after repeated administration, but also that there was a circadian variation in the efficacy of AMK in mice infected with P. aeruginosa. These results suggest that the timing of drug administration should be considered in pharmacotherapy with AMK and that the most appropriate time of administration in mice and nocturnal animals may be in the midlight (resting) phase. They also suggest that the ED50 of AMK against P. aeruginosa infection may be influenced not only by the circadian variation in pharmacokinetics but also by the variations in immune systems suppressed by cyclophosphamide. PMID- 8403070 TI - Tau changes after single nonphotic events. AB - Changes in the free-running period of the circadian rhythms of hamsters occur after single nonphotic events such as a 3-h pulse of running induced by being put in a novel wheel. These changes are mostly in the direction of longer periods, and can exceed 0.2 h; the magnitude of the effect depends on the circadian phase of the pulse. The phase response curves for period changes do not match up with those for phase shifts of the rhythms. Data on free-running rhythms after anisomycin injections and after novelty-induced wheel running in tau mutant hamsters support the view that period changes and phase shifts can occur independently of one another. PMID- 8403071 TI - Double-blind randomized cross-over trial of nocturnal elixir theophylline supplementation of a twice-daily sustained-release theophylline tablet formulation in asthmatic patients. AB - Sixteen asthmatic patients with normal diurnal activity between 05:00 and 23:00 h participated in this randomized, multiple-dose, double-blind, placebo-controlled, crossover study of the pharmacokinetics and efficacy of evening supplementation of a 12-hourly sustained-release theophylline (SRT) regimen with a nonsustained release theophylline (NSRT) formulation. The treatments were Nuelin SA (SRT) every 12 h plus, in the evening, either placebo or an additional dose of Nuelin liquid (NSRT), determined to raise the early morning (0300) plasma theophylline concentration (PTC) to 18 micrograms/ml by using the dose-concentration prediction equation established in a study conducted on healthy volunteers and reported in this journal. The 11-day trial included two 24-h inpatient periods during which PTCs and lung functions (PEF, FEV1, FEF25-75, and FVC) were determined every 2 h. The value of the prediction equation was confirmed when the early morning PTC, after evening supplementation with Nuelin Liquid, was raised nearly to the targeted 18 micrograms/ml. The nocturnal peak-to-trough fluctuation in PTC was larger during additional treatment with Nuelin liquid, but the nocturnal peak-to-trough fluctuation in lung function parameters decreased. Overall, airflow during the early morning hours (0100-0500) significantly improved during this chronotherapeutically optimized treatment of adding an NSRT product to the evening dose of a 12-hourly SRT regimen. PMID- 8403072 TI - Chronopharmacology of albuterol in hospitalized asthmatic children. AB - Eleven children (8-16 years old) hospitalized for acute bronchospasm were included in this investigation. Throughout the study, the children received the standardized course of therapy for hospitalized asthmatics with corticosteroids and albuterol nebulizations. Children receiving ipratropium were excluded from the study. Spirometric measurements, including forced expiratory volume in 1 s (FEV1), were made immediately before and 30 min after each albuterol nebulization over a 24-h period. The well-known temporal changes in FEV1 were observed in patients suffering from nocturnal asthma (NA): basal values were maximal at midday (10 a.m. to 2 p.m.) and lowest in the evening or at night (10 p.m. to 6 a.m.). This 24-h variation in lung function was not found in children without nocturnal exacerbations of their asthma. A 24-h variation was also observed in albuterol-induced bronchodilation in patients with NA: maximal effectiveness occurred at night, and lower effect was obtained with the midday administration. The albuterol-induced increases in FEV1 were not clinically significant in children without nocturnal asthma except when the beta 2-agonist was inhaled between 10 p.m. and 2 a.m. The data suggest that patients with nonnocturnal asthma might have different drug requirements than those with nocturnal symptoms. PMID- 8403073 TI - Circadian rhythms of circulating NK cells in healthy and human immunodeficiency virus-infected men. AB - Antiviral immunity involves NK cells, which circulate rhythmically every 24 hours. We have investigated circadian and 12-hour rhythms in the peripheral count of circulating NK cells in 15 men infected with human immunodeficiency virus (HIV) and 13 healthy controls. We analyzed three phenotypes using double-labeling with monoclonal antibodies and flow cytometry assessment: CD3- CD16+, CD3-CD57+, and CD2+CD3-. A statistical validation of time-dependent differences was achieved if significance (p < 0.05) was validated both with analysis of variance and cosinor. The circadian rhythm had a similar asymmetric waveform for the three phenotypes and is homogeneous on an individual basis. The circulating NK cell count peaked in the early morning and was low at night. A circadian rhythm and a circahemidian harmonic characterized all phenotypes in healthy subjects. We considered two groups of HIV-infected men: those who were asymptomatic (eight) and those with acquired immune deficiency syndrome (AIDS) (seven). Circadian changes in NK cell count were similar in both subgroups and in healthy controls. The circadian pattern was also consistent among individual patients. Asymptomatic HIV-infected men (early-stage disease) exhibited more pronounced 12-hour rhythmicity than did patients with AIDS or controls. The circulation of NK cells does not appear to share the same synchronizer(s) as other circulating T- or B lymphocyte subsets. Thus, HIV infection gradually abolished circadian rhythmicity in circulating T and B cells, whereas it did not disturb that in NK cells. PMID- 8403074 TI - Circadian rhythms and masking: an overview. AB - Masking, as is well known, enables an organism to act immediately and in an appropriate way to changes of the environment, integrating with internally produced rhythmicity. It now appears that masking can be used to cover a far wider range of problems than was originally intended. To separate masking effects from the effects due to an internal oscillator, several techniques have been used. Such protocols, however, like the constant routine protocol, often replace one form of masking by another. The situation becomes even more complex when one realizes that the output of an internal oscillator modifies the input. The question might be asked whether it is possible to study the properties of the internal oscillator in vivo at all. This article attempts to produce a framework for future discussions. PMID- 8403075 TI - In vitro reactions of 2-cyanoethylene oxide with calf thymus DNA. AB - 2-cyanoethylene oxide (CEO) is a direct-acting mutagen and the postulated proximate carcinogenic form of acrylonitrile (AN). We have studied the reactions of CEO with 2'-deoxyribonucleosides and in vitro with calf thymus DNA at pH 7.0 7.5 and 37 degrees C for 3 h. Reaction of CEO with dAdo gave 2 adducts, N6-(2 hydroxy-2-carboxyethyl)-dAdo (N6-HOCE-dAdo) (2% yield) and 1,N6-etheno-dAdo (epsilon-dAdo) (11%); reaction with dCyd resulted in the isolation of 3-HOCE-dUrd (22%); reaction with dGuo gave 7-(2-oxoethyl)-Gua (7-OXE-Gua) (31%) and reaction with dThd yielded 3-OXE-dThd (3%). Structural elucidation of adducts was accomplished by ultraviolet spectroscopy, high-field proton NMR spectroscopy and mass spectrometry. Structural confirmation was provided by an accurate mass measurement technique where diagnostic ions in the electron impact mass spectra of trimethylsilyl derivatives were measured to within 0.0007 atomic mass units. The facile Dimroth rearrangement of 1-HOCE-dAdo to N6-HOCE-dAdo and hydrolytic deamination of a dCyd adduct to 3-HOCE-dUrd is postulated to be catalyzed by the hydroxyl group on the 3-carbon side chain of the adduct. Reaction of CEO with calf thymus DNA yielded (nmol/mg DNA) N6-HOCE-dAdo (2); epsilon-dAdo (11); 3-HOCE dUrd (80); 7-OXE-Gua (110) and 3-OXE-dThd (1). Thus CEO, like its metabolic precursor AN, directly alkylates DNA in vitro but at a much more rapid rate. PMID- 8403076 TI - A comparison of free radical formation by quinone antitumour agents in MCF-7 cells and the role of NAD(P)H (quinone-acceptor) oxidoreductase (DT-diaphorase). AB - Electron paramagnetic resonance (EPR/ESR) spin trapping studies with DMPO revealed that purified rat liver NAD(P)H (quinone-acceptor) oxidoreductase (QAO) mediated hydroxyl radical formation by a diverse range of quinone-based antitumour agents. However, when MCF-7 S9 cell fraction was the source of QAO, EPR studies distinguished four different interactions by these agents and QAO with respect to hydroxyl radical formation: (i) hydroxyl radical formation by diaziquone (AZQ), menadione, 1AQ; 1,5AQ and 1,8AQ was mediated entirely or partially by QAO in MCF-7 S9 fraction; (ii) hydroxyl radical formation by daunorubicin and Adriamycin was not mediated by QAO in MCF-7 S9 fraction; (iii) hydroxyl radical formation by mitomycin C was stimulated in MCF-7 S9 fraction when QAO was inhibited by dicumarol; (iv) no hydroxyl radical formation was detected for 1,4AQ or mitoxantrone in MCF-7 S9 fraction. This study shows that purified rat liver QAO can mediate hydroxyl radical formation by a variety of diverse quinone antitumour agents. However, QAO did not necessarily contribute to hydroxyl radical formation by these agents in MCF-7 S9 fraction and in the case of mitomycin C, QAO played a protective role against hydroxyl radical formation. PMID- 8403077 TI - Effects of nickel ions on polymerase activity and fidelity during DNA replication in vitro. AB - Nickel is a genotoxic carcinogen. However, the mechanisms of nickel-induced genotoxicity are not well understood. We have investigated the effects of Ni2+ ions on DNA polymerase activity and the fidelity of DNA replication in vitro. The effect of Ni2+ on different DNA polymerases is quite variable. The amount of enzyme inhibition and degree of alteration in replication fidelity induced by Ni2+ are dependent both on the polymerase and its associated 3'-5' exonuclease activity. Some polymerases, such as E. coli DNA polymerase I, AMV reverse transcriptase and human DNA polymerase alpha, can utilize Ni2+ as a weak substitute for Mg2+ during DNA replication. Other polymerases are very sensitive to inhibition by Ni2+ and the IC50 can vary by an order of magnitude. T4 polymerase is relatively insensitive to inhibition by Ni2+, although the sensitivity is enhanced in the absence of added Mg2+, and Ni preferentially inhibits the 3'-5' exonuclease function of T7 DNA polymerase. The fidelity and processivity of DNA polymerases may be either increased or decreased by Ni ions in a polymerase dependent manner. The inhibition DNA polymerase activity and altered replication fidelity may contribute significantly to Ni-induced mutagenesis and genotoxicity in vivo. PMID- 8403078 TI - Effects of polychlorinated dibenzofurans on compounds in hepatic DNA of female Sprague-Dawley rats: structure dependence and mechanistic considerations. AB - Previous work indicated that covalent age-dependent DNA modifications of endogenous origin termed I-compounds may represent useful biomarkers for tumor promotion/carcinogenesis, as various tumor promoters/carcinogens, including 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and phenobarbital, reduce rat I compound levels in liver, the target organ. The present study addressed the question as to whether polychlorinated dibenzofurans (PCDFs), which are related to TCDD and its congeners with regard to their toxic and biochemical properties, would also affect hepatic I-compound patterns and levels, and whether such effects would be chemical structure-dependent. Female Sprague-Dawley rats were treated once a week with a single dose (100 micrograms/kg) of 1,2,3,7,8 pentachlorodibenzofuran (1,2,3,7,8-PeCDF), 1,2,4,7,8-PeCDF, 2,3,4,7,8-PeCDF, or 2,3,4,6,7,8-hexachlorodibenzofuran (2,3,4,6,7,8-HeCDF) for 4 weeks and liver DNA was analyzed at the end of the last week by 32P-postlabeling assay. No carcinogen DNA adducts were detected; however, levels of both non-polar and polar I compounds were reduced in a structure-dependent manner. Potencies increased in the order, control (100%, 122 modifications in 10(9) DNA nucleotides = 1,2,4,7,8 PeCDF (104%) < 1,2,3,7,8-PeCDF (80%) < 2,3,4,7,8-PeCDF (61%) and 2,3,4,6,7,8 HeCDF (61%). Structure-activity relationships for total I-compounds, therefore, paralleled those reported for Ah receptor agonist activity, i.e., compounds that exhibit high cytosolic Ah receptor binding affinities and are also potent inducers of aryl hydrocarbon hydroxylase activity (1,2,3,7,8-PeCDF, 2,3,4,7,8 PeCDF, and 2,3,4,6,7,8-HeCDF) were active, while 1,2,4,7,8-PeCDF, which is a less potent Ah receptor agonist, was inactive. Polar I-compounds responded to a greater extent than did non-polar ones and, in general, individual I-compounds were affected differentially, thus decreased formation or increased removal of I compounds played a role in the observed effects of the toxins on DNA. It is proposed that Ah receptor-mediated enzyme induction, particularly of cytochrome P450, is involved in reduced hepatic I-compound formation and that subnormal I compound levels may contribute to tumor promotion. PMID- 8403079 TI - Correlation of metabolism, covalent binding and toxicity for a series of bromobenzene derivatives using rat liver slices in vitro. AB - For many acute-acting chemicals, toxic responses observed in vivo correlate strongly with metabolic activation and macromolecular covalent binding (CVB) observed in vitro and often in vivo; bromobenzene (BB) is a classic example of this behavior. Substituent groups modulate the toxicity of bromobenzene in vivo and in liver slices cultured in vitro in parallel fashion [Fisher, R., Hanzlik, R.P., Gandolfi, J.A., and Brendel, K. (1992), In Vitro Toxicology, 4, 173-186]. In the present study we used the liver slice system to examine the relationship between toxicity, metabolism and covalent binding amongst a series of [3H/14C] dual labelled BB derivatives including (in order of increasing hepatotoxicity) o bromoanisole (BA), o-bromotoluene (BT), o-bromobenzonitrile (BBN), BB and o dibromobenzene (DBB). Among these congeners apparent relative rates of metabolism varied only 4-fold, but the most extensively metabolized compounds were the least toxic. CVB varied 7-fold across the series, and those compounds which bound the most frequently were the most toxic. For each compound the relative binding index (RBI = pmol bound/nmol metabolized) and the average retention of tritium relative to carbon-14 in the CVB fraction were constant throughout the 24 h incubations, suggesting that the metabolic profile of each compound remained constant with time. The RBI values, however, did not reflect relative toxicity as well as total CVB values. The T/C ratios of the CVB residues varied from 0.36 (for BA) to 0.81 (for BBN), indicating that ortho-substitution on BB exerts important qualitative as well as quantitative effects on overall metabolism and reactive metabolite formation. The finding that relative toxicity among a series of bromobenzene congeners is paralleled by their relative covalent binding measured in the same system in which toxicity is assessed adds support to the hypothesis that covalent binding contributes to the observed toxicity, rather than merely being a correlated epiphenomenon. PMID- 8403080 TI - N-methyl-4-phenylpyridinium (MPP+) potentiates the killing of cultured hepatocytes by catecholamines. AB - The role of catecholamines in the toxicity of MPTP (N-methyl-4-phenyl- 1,2,3,6 tetrahydropyridine) was explored. The killing of cultured hepatocytes by dopamine and 6-hydroxydopamine was enhanced following inhibition of glutathione reductase by 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), a manipulation known to sensitize such cells to an oxidative stress. The participation of activated oxygen species in the cell injury under such circumstances was shown by the ability of catalase and the ferric iron chelator deferoxamine to protect the hepatocytes. The toxicity of catecholamines was also potentiated by the mitochondrial site I (NADH dehydrogenase) inhibitor rotenone. MPP+ (N-methyl-4-phenyl-pyridinium), the putative toxic metabolite of MPTP is also a site I inhibitor. Incubation of hepatocytes with MPP+ similarly potentiated the toxicity of 6-hydroxydopamine, dopamine, and norepinephrine under conditions where MPP+ alone or catecholamines alone did not kill cells. Hepatocytes that had accumulated dopamine from the medium were killed by a subsequent exposure to MPP+ in the absence of a catecholamine in the medium. Hepatocytes that had not been pretreated with dopamine were not affected by the subsequent exposure to MPP+. These data indicated that catecholamines render hepatocytes more susceptible to the toxicity of MPP+ and suggest that the presence of catecholamines in specific neurons in the brain may be related to the selective neurotoxicity of MPTP. PMID- 8403081 TI - A chemical hypothesis for arsenic methylation in mammals. AB - The toxicology of arsenic is complicated by its ability to convert between oxidation states and organometalloidal forms. These processes cause differences in the relative tissue-binding affinities of the various arsenic species, and they determine both the intoxication and the detoxification mechanisms. In this review, a chemical hypothesis of arsenic biomethylation is developed from an examination of data and observations presented by researchers who conducted numerous in vivo and in vitro experiments. It is likely that a combination of pathways is actually used during methylation of arsenic in vivo, and that the principal mechanism depends on various factors affecting the cellular environment. Despite these uncertainties, several observations can be made: (i) glutathione (GSH) is required for reduction of arsenic(V) to arsenic(III) species in preparation for enzyme-catalyzed oxidative methylation; (ii) GSH is not involved in monomethylation once arsenite is formed, but GSH is involved in dimethylation by reducing methylarsonic acid [MMA(V)] to methylarsonous acid [MMA(III)]; (iii) GSH is also required in the methylation of arsenic by stabilizing the reductive nature of the cell; (iv) a different methyltransferase is used in each methylation step; (v) dithiols (either a cofactor or the methyltransferases) are required for both mono- and dimethylation and (vi) where dithiols are involved, oxidative methylation reduces the stability of the arsenic sulfur complex and permits dissociation of the arsenic species. This lower affinity of the pentavalent organoarsenic species for dithiols is part of the reason why methylation of arsenic can be a detoxification mechanism when the As(III) intermediates are not permitted to accumulate. PMID- 8403082 TI - Saponin and sapogenol. XLVIII. On the constituents of the roots of Glycyrrhiza uralensis Fischer from northeastern China. (2). Licorice-saponins D3, E2, F3, G2, H2, J2, and K2. AB - Following the characterization of licorice-saponins A3 (2), B2 (3), and C2 (4), the chemical structures of licorice-saponins D3 (5), E2 (6), F3 (7), G2 (8), H2 (9), J2 (10), and K2 (11), seven of the ten oleanane-type triterpene oligoglycosides isolated from the air-dried roots of Glycyrrhiza uralensis Fischer collected in the northeastern part of China, were investigated. On the basis of chemical and physicochemical evidence, the structures of licorice saponins D3, E2, F3, G2, H2, J2, and K2 have been determined to be expressed as 3 beta-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucuronopyranosyl(1-- >2)-beta-D glucuronopyranosyloxy]-22 beta-acetoxyolean-12-en-30-oic acid (5), 3-O-[beta-D glucuronopyranosyl(1-->2)-beta-D- glucuronopyranosyl]glabrolide (6), 3-O-[alpha-L rhamnopyranosyl(1-->2)-beta-D-glucuronopyranosyl(1--> 2)-beta-D glucuronopyranosyl]-11-deoxoglabrolide (7), 24-hydroxyglycyrrhizin (8), 3-O-[beta D-glucuronopyranosyl(1-->2)-beta- D-glucuronopyranosyl]liquiritic acid (9), 24 hydroxy-11-deoxoglycyrrhizin (10), and 3 beta-[beta-D- glucuronopyranosyl(1-->2) beta-D-glucuronopyranosyloxy]-24-+ ++hydroxyoleana- 11,13(18)-dien-30-oic acid (11), respectively. PMID- 8403083 TI - Saponin and sapogenol. XLIX. On the constituents of the roots of Glycyrrhiza inflata Batalin from Xinjiang, China. Characterization of two sweet oleanane-type triterpene oligoglycosides, apioglycyrrhizin and araboglycyrrhizin. AB - Two sweet oleanane-type triterpene oligoglycosides named apioglycyrrhizin and araboglycyrrhizin were isolated from the air-dried roots of Glycyrrhiza inflata Batalin, collected in Xinjiang province (Shinkyo-Kanzo in Japanese), together with glycyrrhizin (3), licorice-saponins A3 (8), G2 (10), and H2 (11) and known flavonoid glycosides. On the basis of chemical and physicochemical evidence, the structures of apioglycyrrhizin and araboglycyrrhizin have been determined to be expressed as 3-O-[beta-D-apiofuranosyl(1-->2)-beta-D- glucuronopyranosyl]glycyrrhetic acid (1) and 3-O-[alpha-L- arabinopyranosyl(1- >2)-beta-D-glucuronopyranosyl]glycyrrhet ic acid (2), respectively. During the course of these studies, it has been found that the hydroxyl groups in the oligosaccharide moiety of the glucuronide saponins may be partially methylated by prolonged treatment with diazomethane in methanol. The sweetness of the saponins hitherto isolated from various Glycyrrhizae Radix has been examined and a structure-sweetness relationship, as compared with glycyrrhizin, has been found. PMID- 8403084 TI - Isolation and characterization of seven lyso platelet-activating factors and two lyso phosphatidylcholines from the crude drug "suitetsu" (the leech, Hirudo nipponica). AB - Nine lyso glycerophospholipids were isolated in the pure state from the crude drug "Suitetsu", which is the dried body of the leech, Hirudo nipponica (Hirudidae). They were identified as 1-O-hexadecyl-(1), 1-O-octadecyl-(2), 1-O tetradecyl-(3), 1-O-9-cis-hexadecenyl-(4), 1-O-hexadecanoyl-(5), 1-O-pentadecyl (6), 1-O-15-methylhexadecyl-(7), 1-O-octadecanoyl-(8) and 1-O-heptadecyl-sn glycero-3-phosphocholine (9). Two of them (5 and 8) are lysophosphatidylcholines and the other seven are lyso platelet-activating factors. One of them has an alkenyl carbon chain. PMID- 8403085 TI - Studies of HIV-1 protease inhibitors. I. Incorporation of a reduced peptide, simple aminoalcohol, and statine analog at the scissile site of substrate sequences. AB - Inhibitors of the protease of human immunodeficiency virus type-1 (HIV-1) were designed and synthesized. A reduced peptide, simple aminoalcohol, and statine analog, 4-amino-3-hydroxy-5-phenylpentanoic acid (AHPPA), were inserted at the scissile site of substrate sequences of HIV-1 protease. While both reduced peptides and simple aminoalcohol derivatives were weak inhibitors, the peptides containing AHPPA demonstrated moderate inhibitory activity. The more potent alcohol configuration of AHPPA is (R), which is opposite to the configuration in potent inhibitors of other aspartic proteases. In particular, compound 28 ((3R,4S)-4-(N-tert-butoxycarbonyl- L-glutaminyl-L-asparaginyl)amino-3-hydroxy-5 phenylpentanoic acid 2'-methylbutylamide) had a Ki of 0.36 microM and exhibited excellent enzyme specificity. PMID- 8403086 TI - Studies of HIV-1 protease inhibitors. II. Incorporation of four types of hydroxyethylene dipeptide isosteres at the scissile site of substrate sequences. AB - Human immunodeficiency virus type 1 (HIV-1) protease inhibitors containing four types of hydroxyethylene dipeptide isosteres were designed and synthesized. These inhibitors consist of eight stereoisomers of phenylalanylproline (Phe-psi[H.E.] Pro), four stereoisomers of phenylalanylalanine [Phe-psi[H.E.]-Ala), and one stereoisomer each of phenylalanylglycine (Phe-psi[H.E.]-Gly) and cyclohexylalanylalanine (Cha-psi[H.E.]-Ala) hydroxyethylene dipeptide isosteres. For the synthesis of the latter two isosteres, a newly developed synthetic method for gamma-lactone was applied. The inhibitory activities of these peptides were evaluated by cleavage assay of partially purified gag proteins or purified synthetic peptide. Of the inhibitors examined, compounds 2c (Z-Asn-(2S,3R,4S,5S) Phe-psi[H.E.]-Pro-NHB(un); Bu(n) = n-butyl, Ki = 0.50 microM), 21a (Z-Asn (2R,4S,5S)-Phe-psi[H.E.]-Ala- NHBu(n), Ki = 0.34 microM) and 23 (Z-Asn-(2R,4S,5S) Cha-psi[H.E.]-Ala- NHBu(n), Ki = 0.46 microM) were moderately potent inhibitors. The results revealed that the alkyl substituent at C2 is essential, and the stereochemistry of the hydroxyethylene dipeptide isosteres greatly affected their inhibitory activities. PMID- 8403087 TI - Syntheses and inhibitory effects on gastric lesions of 4-guanidinomethylbenzoic acid arylamides. AB - A novel series of 4-guanidinomethylbenzoic acid (GMBA) arylamides was synthesized. Several showed more potent inhibitory effects on stress-induced gastric lesion in rats than cetraxate. We selected 4-guanidinomethylbenzoic acid (2'-ethoxycarbonyl)phenylamide 3 for further pharmacological assessments because it had low toxicity. Compound 3 showed significant inhibitory effects on stress-, HCl-ethanol- and indomethacin-induced gastric lesions and gastric secretion, the ED50 values being 34.4, 45.0 and 23.0 mg/kg (p.o.) and 240 mg/kg (i.d.), respectively. Furthermore, this compound restored the reduction of gastric mucus caused by the stress-loading and inhibited compound 48/80-induced ulcer. PMID- 8403088 TI - Comparison of cytoprotective effects of saponins isolated from leaves of Aralia elata Seem. (Araliaceae) with synthesized bisdesmosides of oleanoic acid and hederagenin on carbon tetrachloride-induced hepatic injury. AB - Glycosylations of 3-O-(2,3,4-tri-O-acetyl-alpha-L-rhamnopyranosyl(1-->2)- 3,4-di O-acetyl-alpha-L-arabinopyranosyl)-23-O-acetylhederageni n (15) with mono- (16), di- (17) and trisaccharide bromide (18) gave the bisdesmoside peracetates 19, 20 and 22, respectively, which were treated with 5% KOH in MeOH to give the bisdesmosides 25-27. Hydrolysis of the glycosides 6 and 9 having beta-D glucopyranose as a terminal sugar component with beta-glucosidase in acetate buffer (pH 4.7) gave compounds 28 and 29, respectively. Cytoprotective effects of the synthesized triterpenoidal saponins against CCl4-induced hepatic injury were compared with those of saponins isolated from the leaves of Aralia elata Seem. (Araliaceae) using isolated hepatocytes from rat liver. Although the monodesmosides 1-4 having neutral sugar components only at the O-3 position on the aglycones showed no cytoprotective effect, bisdesmosides having sugar components at both the O-3 and O-28 positions on the aglycones had potent effects, even when the species of the sugar components were different. The bisdesmosides 10, 11, and 27 having five monosaccharides in the molecules exhibited the most potent cytoprotective effects. PMID- 8403089 TI - Studies on RA derivatives. V. Synthesis and antitumor activity of Ala2-modified RA-VII derivatives. AB - A number of RA-VII derivatives having various amino acids including proline (6), pipecolic acid (11), norvaline (12), ornithine (14), aspartic acid (15) and methionine (20) in place of Ala2 have been synthesized from RA-X methyl ester (3) and evaluated for cytotoxicity to P388 leukemia and KB cells in vitro. Comparison of the cytotoxicity of these compounds suggests that the polarity and the length of the 2nd amino acid residue affect the activity. An NMR study revealed that, in solution, 6 and 11 are locked in one conformational state, corresponding to conformer A of RA-VII. PMID- 8403090 TI - Structures and solid state tautomeric forms of two novel antileukemic tropoloisoquinoline alkaloids, pareirubrines A and B, from Cissampelos pareira. AB - Two novel tropoloisoquinoline alkaloids, Pareirubrines A and B, have been isolated as antileukemic substances from Cissampelos pareira (Menispermaceae), together with the same skeleton alkaloids, grandirubrine and isoimerubrine. Their structures were elucidated by nuclear magnetic resonance (NMR) studies, and their solid state tautomeric forms were examined by X-ray crystallographic analysis. PMID- 8403091 TI - Effects of oleic acid/propylene glycol on rat abdominal stratum corneum: lipid extraction and appearance of propylene glycol in the dermis measured by Fourier transform infrared/attenuated total reflectance (FT-IR/ATR) spectroscopy. AB - Fourie transform infrared/attenuated total reflection analysis demonstrated that the absorbance intensity of C = O stretching bands, which reflect the amounts of lipids in the stratum corneum, decreased with an increase in the duration of skin treatment with 0.15 M oleic acid/propylene glycol (PG) system, suggesting that the oleic acid/PG system induced the lipid extraction, which was followed by a reorganization of the stratum corneum structures. The spectral peaks which originated from the PG molecule were detected in dermal tissues after 30 min of treatment of the stratum corneum with the same system. This observation suggested that the reorganization of the lipid domains due to the lipid extraction by the oleic acid/PG system helped the PG molecules enter the dermal tissues. It was also suggested that an effective volume within the stratum corneum for solutes and/or solvents which could penetrate through the inter-, and/or intracellular routes could be altered in conjunction with the structural changes of the lipids. PMID- 8403092 TI - Inclusion complexation of p-hydroxybenzoic acid esters with 2-hydroxypropyl-beta cyclodextrins. On changes in solubility and antimicrobial activity. AB - To obtain a transparent and effective solution of p-hydroxybenzoic acid esters (parabens), the use of 2-hydroxypropyl-beta-cyclodextrins (2-HP-beta-CyDs) as solubilizers with different degrees of substitution (D.S.) was surveyed. 2-HP beta-CyDs significantly increased the aqueous solubility of four kinds of parabens (methyl < ethyl < propyl < butyl esters), where the solubilizing ability decreased with an increase in the D.S. of the 2-hydroxypropyl group in beta-CyD. The antimicrobial activity of the parabens tended to decrease by complexation with 2-HP-beta-CyDs. However, the activity could be maintained by lengthening the alkyl chain of the parabens. 1H- and 13C-nuclear magnetic resonance and circular dichroism spectroscopic studies suggest that the hydrophobic alkyl moiety of butyl paraben is preferably included in the cavity, and the phenol group extrudes from the cavity. The present results suggest that a suitable combination of 2-HP beta-CyDs and hydrophobic, longer alkyl parabens is useful for the preservation of liquid formulations. PMID- 8403093 TI - Lipid A and related compounds. XXVII. An efficient synthesis of D-galactosamine-4 phosphate analogs of lipid A via a novel key intermediate. AB - A new methodology for chemical differentiation of one amino and four hydroxyl groups of D-galactosamine derivatives and its application for the synthesis of D galactosamine-4-phosphate analogs of lipid A are described. Preliminary examination of biological activity revealed that the synthetic monosaccharides show mitogenic activity. PMID- 8403094 TI - Conformation of tropolone ring in antileukemic tropoloisoquinoline alkaloids. AB - Conformational analysis of antileukemic tropoloisoquinoline alkaloids isolated from Cissampelos pareira was conducted by thermodynamic proton nuclear magnetic resonance (1H NMR) studies. The line-broadening of one of methoxy methyl signals can be explained by the tropolone ring-puckering process. Analysis by dynamical simulated annealing and modified neglect of differential overlap (MNDO) calculations also supported puckering of tropolone ring system. PMID- 8403095 TI - [The value of hybridization-in-situ technique in the diagnosis of condyloma acuminatum]. AB - By using 35S-dCTP labelled HPV16 probe, the HPV DNA sequence in 32 cases of condyloma acuminatum and suspicious condyloma acuminatum of vulva and vagina as well as 18 cases of papillomas at different anatomical sites were detected. The results showed that the HPV DNA sequences were positive in all 25 cases of condyloma acuminata which were typical both clinically and pathologically and in 6/7 of the suspicious cases; whereas in only 1/18 of the papillomas at various sites were they positive. Thus, nucleic acid hybridization-in-situ technique combined with histopathology seemed to be of great value for the diagnosis and differential diagnosis of condyloma acuminatum. PMID- 8403096 TI - [Signal transduction study on the growth regulation of cells from human giant cell lung carcinoma in vitro]. AB - The signal transduction involved in growth regulation of cells originally from a human giant cell lung carcinoma(PG) was investigated. The purinergic receptor agonists, ATP and its analogues, as well as bombesin all played a significant growth regulatory role on PG cells. ATP showed a growth inhibitory effect while bombesin showed a growth stimulatory effect on PG cells in vitro. Further investigation showed that ATP and bombesin activated phosphatidylinositol turnover/calcium mobilization signal transduction pathways in PG cells. and increased production of inositol-(1,4,5) trisphosphate and mobilization of intracellular free calcium, in which ATP and bombesin effects were dose dependent. The purinergic receptor on PG cells was characterized by P2y subtype according to the order of PG cell responses to a series of ATP analogues. Pretreatment with cholera toxin showed different effects on the functions of ATP and bombesin, suggesting that the major difference between ATP and bombesin on signal transduction pathways may be at the G protein level. PMID- 8403097 TI - [New trends in the research on infiltrating lymphocytes]. PMID- 8403098 TI - [A comparative study of six different staining methods on neurofibrillary tangles and senile plaques]. AB - Six different staining methods were evaluated for demonstrating SPs and NFTs by using serial sections of brain tissue collected from aged patients with or without senile dementia. The methods used were as follows: the Bodian, Modified Beilschowsky, Periodic acid methenamine silver (PAMS), New methenamine silver (MS), PAS and Congo Red. SPs and NFTs were demonstrated in the cerebral cortex of aged patients with or without dementia. Among these 6 methods Bodian gives a more reliable result in demonstrating tangles and plaques of the classic and primitive types but not the diffuse type. The MS method clearly demonstrated all the 3 types of SPs particularly the diffuse plaque, but could not show the NFTs. The result indicates that combination of Bodian and MS methods seemed to be the most reliable method for routine neuropathological investigation. PMID- 8403099 TI - [A technique for bone marrow biopsy specimen preservation]. AB - The rapidly frozen bone marrow specimen was displaced in iced acetone at low temperature first and was embedded in low-temperature glycol methacrylate resin subsequently which was then ready for cryostat section preparation. Since there was no routine fixation required, so that the morphology features will be well preserved for further immunohistochemical and histochemical studies. By this technique, the enzyme activity and localization site will also be well preserved, particularly the lymphoid cell surface antigens, and it is also valuable for retrospective study of biopsy specimens. PMID- 8403100 TI - [Localization and significance of lysozyme in tuberculosis]. AB - The relationship between expression of lysozyme and tuberculosis lesions was studied in 132 patients. The positive rate of lysozyme expression in 132 tuberculosis cases was 83.3%. Lysozyme was mainly distributed intracellularly, and the positive rates in different types, i.e. in cases with lesions of caseous necrosis, cellular nodes, and fibroid nodes were 90.1%, 79.3% and 63.3% respectively (P < 0.05 between lesions of caseous necrosis and fibroid nodes). The positive rate of lysozyme in cases of infection associated with L-form tubercule bacillus was 64.4%, which was higher in cases with positive L-form antigen than that in antigen negative cases (P < 0.025). The results indicated that expression of lysozyme was rather active, and might be considered as a marker for the prognosis of tuberculosis treatment. PMID- 8403101 TI - [Histopathology and ultrastructural changes of the musculature and nerves during methamidophos toxication in chicken]. AB - After exposure to methamidophos (Via digestive or respiratory tract or just by skin contamination) for 1, 2, 3, 4, 8 and 12 weeks respectively, ultrastructure of the myofibrils in chicken with methamidophos toxication via the digestive tract showed mild degeneration in the first week and serious degeneration by the end of the fourth week. Light microscopy of the muscle-fibril showed atrophy and necrosis in the third week, including both myopathic atrophy and neuropathic atrophy. Recovery of the lesions took place from the 8th to the 12th week (Light or elcetion microscopy). Those chickenstaking immunosuppressive drug simultaneously with methamidophos exposure (via digestive tract) showed less degeneration of the myofibrils and nerves. Ultrastructure of the myofibril in chickens with methamidophos intoxication through the respiratory tract also developed less degeneration of myofibrils and nerves than those obtained through digestive intoxication. There was no lesion obtained in chicken taking methamidophos via the respiratory tract coexistently with the immunosuppressive drug or just by skin contamination. PMID- 8403102 TI - [Morphometric study on the sudanophilic lesions of arterial intima in young adults]. AB - Sudanophilic lesions (SL) of the aortic intima and right coronary artery (RCA) in 129 and 113 cases respectively were calculated by image analyses. The age ranged from 15 to 34. The probability-of-occurrence-map of aortic SL was higher at the area around the orifices of the arterial branching sites and at the posterior wall of the aorta, lower at the lateral wall and least in the ventrical wall. The prevalence of SL occurred in the RCA was lower which was also less in severity and showed a positive correlation with the lesion developed at the aortic intima. PMID- 8403103 TI - [An electron microscopic observation on spike foci of human epileptic brains]. AB - Twenty cases of ECOG spike foci in epilepsy originally recorded by flaky silver electrodes on the surface of cortices and then to be removed surgically were studied under transmission electron microscope. The ultrastructural changes of the spike foci seemed to be identical in all cases although there were differences in etiologies, locations and seizure types. The presynaptic terminals of axospinal asymmetric synapses were obviously swollen anyhow, the postsynaptic spines were relatively unaffected. The synaptic vesicles were decreasing in number and to be accumulated predominantly at the presynaptic membrane which indicated that these excitatory changes of synapses may play an important role in the mechanism of epileptic abnormal electric activity and the development of epilepsy. Additionally, the ultrastructural changes and the cytopathologic significance of neurons, astrocytes and capillaries are also discussed. PMID- 8403104 TI - [Histopathological study of intracranial germinoma]. AB - Eighteen cases of intracranial germinoma (16 male and 2 female, age 2.5-30 years, average 16.7) were studied. Morphologically, the pathological changes in 18 cases were similar to those of testis seminoma or ovarian dysgerminoma. Among them, two were accompanied with choriocarcinoma and one with embryonal cancer. No cytoplasmic processes was detected by silver stain, but strongly positive to PAS stain. All of the 18 cases in this series as well as another 8 cases of testis seminoma and ovarian dysgerminoma used as controls gave a positive reaction in alkaline phosphatase stain. Among the 18 cases of germinoma, except 3 out of 18 were CEA positive, otherwise, all were CEA, beta-HCG, SP1, AFP and GFAP negative. The syncytiotrophoblasts in the choriocarcinoma and embryonal cancer mentioned before were beta-HCG and SP1 positive, and additionally, the embryonal cancer also showed AFP positive. All these results support the hypothesis that both intracranial (extragonadal) and gonadal germinoma have a common cell origin. PMID- 8403105 TI - [Immunohistochemical study in astrocytoma]. AB - Seventy-four cases of human astrocytoma were studied with ABC method for GFAP and vimentin. Data indicated that all of the specimens in this series contained two different intermediate filament proteins: GFAP and vimentin. There was a negative correlation between the degree of anaplasia and the number of GFAP-positive cells, but no definite relationship was seen between the degree of anaplasia and the number of vimentin-positive cells. The ratio of V/G might be considered useful in expressing the histological differentiation of astrocytoma. Generally, two immunostainings are used in demonstrating GFAP and vimentin: one for intense perikaryo-staining and the other for cell processes. Both staining patterns are closely related to the histological type but not to the histological grade. Besides, there was a negative correlation between the cell density and the number of GFAP-positive cells. PMID- 8403106 TI - [Morphology, immunohistochemistry and differential diagnosis of 70 cases of adenocarcinoma of uterine cervix]. AB - Seventy cases of cervical adenocarcinoma were classified into five types, namely: mucinous adenocarcinoma, endometrioid adenocarcinoma, mixed adenosquamous carcinoma, papillary adenocarcinoma and clear cell carcinoma. According to the morphological features, the results of mucin and immunohistochemical stainings, the histogenesis of cervical endometrioid adenocarcinoma and the differential diagnosis between cervical adenocarcinoma and endometrioid adenocarcinoma are discussed. The significance of vimentin staining for the differential diagnosis is also evaluated. PMID- 8403107 TI - Observations on structural features and characteristics of biological apatite crystals. 5. Three-dimensional observation on ultrastructure of human enamel crystals. AB - In a series of studies to investigate the structural features of the biological crystals, such as the tooth and bone, using an electron microscope, we examined the ultrastructure of the enamel, dentin, and bone crystals at near atomic resolution and showed the configuration of the hydroxyapatite structure through the cross and longitudinal sections of the crystals. Thereafter, based on the results of the observations by the authors of the ultrastructure of the tooth and bone crystals, thinking that it might be possible to conduct direct three dimensional observation of the configuration composing the unit cell of the hydroxyapatite crystals, we conducted a study on this. These results indicated that it was possible to sterically observe the configuration of the hydroxyapatite structure composing the enamel crystal. The materials used for this study were the middle layer of the noncarious enamel from the freshly extracted human erupted permanent molars. The small cubes of the enamel were fixed in glutaraldehyde and osmium tetroxide and embedded in epoxy resin using the routine methods. The ultrathin sections were cut with a diamond knife without decalcification and were examined with the HITACHI H-9000 H type transmission electron microscope operated at 300 kV. Each crystal was observed at an initial magnification of 500,000 times and at the final magnification of 10,000,000 times and over. We sincerely believe that the electron micrographs shown in this report are the first to show three-dimensionally the configuration of the hydroxyapatite structure composing the crystal in the cross and longitudinal sections of an enamel crystal. PMID- 8403108 TI - Observations on structural features and characteristics of biological apatite crystals. 6. Observation on lattice imperfection of human tooth and bone crystals. I. AB - In a series of studies to investigate the basic structural features and characteristics of the biological apatite crystals, using an electron microscope, we examined the ultrastructure of the human enamel, dentin, and bone crystals at near atomic resolution and showed the configuration of the hydroxyapatite structure through the cross and longitudinal sections of the crystals. Subsequently, based on the results of the observations by the authors of the ultrastructure of the tooth and bone, using the same approach, we have been able to directly examine the images of the lattice imperfections in the human enamel, dentin, and bone crystals, such as the point defect structures and dislocations in the crystals. In this report, we describe the image of the point defect structures and line defect structures obtained, using the same approach from the sections of the human enamel, dentin, and bone crystals. The materials used for this study were the noncarious enamel and dentin from the freshly extracted human erupted lower first molars, and bone tissue obtained from the alveolar compact bone. The small cubes of the material were fixed in glutaraldehyde and osmium tetroxide and embedded in epoxy resin using the routine methods. The ultrathin sections were cut with a diamond knife without decalcification. The sections were examined with the HITACHI H-800 H and H-9000 types of transmission electron microscopes operated at 200 kV and 300 kV. Each crystal was observed at the initial magnification of 300,000-500,000 times and at the final magnification of 10,000,000 times and over. We sincerely believe that the electron micrographs shown in this report are the first to show the images of the lattice imperfections in the human enamel, dentin, and bone crystals, such as the point defect and line defect structures, at near atomic resolution. PMID- 8403109 TI - The maintenance of competence programme (MOCOMP) PMID- 8403110 TI - Continuing medical education practices of community-based and university affiliated anaesthetists in Ontario. AB - To remedy the lack of information about the continuing medical education (CME) practices of anaesthetists, we designed a survey to define and compare the CME activities of specialist anaesthetists in community-based and university affiliated practices: 463 members of the Canadian Anaesthetists' Society in the Province of Ontario (263 community-based and 200 university-affiliated (University of Toronto) anaesthetists). Data from 304 (65.6%) respondents (172 community-based and 132 university-affiliated anaesthetists) were analyzed by non parametric analysis (statistical significance P < 0.05). Most respondents spent between two to four hours per week on CME activities. Journal reading was the most commonly used method to obtain CME and was perceived to be the most efficient of the methods surveyed (P < 0.05). Formal teaching, including seminars, workshops, and annual society meetings, although the second most commonly used technique to obtain CME, was considered as effective as journal reading. Instructional media techniques were the least commonly used and considered the least effective (P < 0.05). Most community-based and university affiliated anaesthetists obtained CME by a variety of techniques; of all respondents, 77% have no formal method of assessing their learning needs and 88% would consider participation in a formalized learning needs assessment programme. PMID- 8403111 TI - A clinical assessment of desflurane anaesthesia and comparison with isoflurane. AB - In 48 randomly assigned ASA I adult patients undergoing elective orthopaedic procedures, we compared the pharmacodynamics of desflurane (DF) and isoflurane (IF), and their pharmacokinetics during rapid induction of deep anaesthesia (via face mask, to 1.5-2 MAC, after thiopentone), maintenance of anaesthesia at 1.25 MAC, and emergence therefrom. During induction, laryngeal reactions ranging from mild crowing to laryngospasm occurred more frequently with DF than with IF (15/24 DF, 5/24 IF; P < 0.05) and was more severe (9/24 DF, 1/24 IF, excluding the mildest form, P < 0.05). As a result, induction of anaesthesia was not accomplished faster with DF, in spite of a faster equilibration between exhaled and inhaled concentrations. Emergence from DF was more rapid and less complicated by delirium. Pharmacokinetically, the exhaled concentration of DF reached 90% of the inhaled concentration within five minutes of induction, whereas that of IF lagged behind and remained 25% below the inhaled concentration (1 vs 1.34 +/- 0.05) even one hour after induction. Premature ventricular contractions did not occur in any patient even during periods of difficulty with the airway and oxygen desaturation. It is concluded that DF is a safe anaesthetic, pharmacokinetically superior to IF but clinically inferior for induction of anaesthesia via a face mask. Because of the fast equilibration, the exhaled concentration of DF can be controlled more precisely by the dial setting of the vaporiser. PMID- 8403112 TI - Amrinone, in combination with norepinephrine, is an effective first-line drug for difficult separation from cardiopulmonary bypass. AB - A crucial element for weaning patients from cardiopulmonary bypass (CPB) rests on the selection of an appropriate therapeutic regimen. Amrinone, a phosphodiesterase III inhibitor, combines inotropic support with pulmonary and systemic vasodilatation, without increasing heart rate (HR) or myocardial oxygen consumption. These characteristics should be useful in the failing heart during weaning from CPB. Nineteen patients were included in this prospective, open labelled, phase IV study when systolic blood pressure (DPAP) > 15 mmHg or central venous pressure (CVP) > 15 mmHg, during progressive separation from CPB. At that moment, CPB flow was increased to alleviate heart failure and amrinone administered as a bolus (0.75 mg.kg-1) followed by an infusion (10 micrograms.kg 1.min-1). Weaning from CPB was then resumed and haemodynamic variables (SBP, DPAP, CVP and HR) were compared with those measured at CPB flow when failure had first occurred. Failure to wean from CPB occurred at 57 +/- 28% of full pump flow. After the amrinone bolus, DPAP and CVP decreased by 20% and 21% respectively. Subsequently, 16 patients required the infusion of norepinephrine (4-8 micrograms.min-1) to maintain a SBP > 80 mmHg. Heart rate remained unchanged after the bolus of amrinone, after separation from CPB, and no arrhythmias were noted. Successful weaning from CPB was possible 12 +/- 8 min after the amrinone bolus. Weaning resulted in a cardiac index similar to that measured pre-bypass. Amrinone is rapidly effective during weaning from CPB and, in combination with norepinephrine, provides the necessary inotropic support during this unstable period. PMID- 8403113 TI - Obstetrical analgesia in a parturient with antithrombin III deficiency. AB - Antithrombin III (AT III) deficiency is a rare inherited disorder which predisposes patients to thrombotic complications. Anticoagulation is necessary to prevent recurrent thrombosis and high doses of heparin are often required. Anticoagulation complicates analgesia in parturients with the condition. We describe such a patient, in pre-term labour, who was successfully managed during labour with intravenous nalbuphine and inhaled nitrous oxide (N2O). PMID- 8403114 TI - Bradycardia during cold ocular irrigation under general anaesthesia: an example of the diving reflex. AB - A case of bradycardia is reported which was precipitated by cold normal saline applied to the eye during general anaesthesia. The history and physiology of the diving reflex is discussed and we believe that these data suggest that this patient's bradycardia was induced by the diving reflex, and not by the oculocardiac reflex. PMID- 8403115 TI - Blunt cardiac injury and intraoperative hypoxaemia. AB - A 28-yr-old man sustained blunt chest trauma in a motor vehicle accident. Severe intraoperative hypoxaemia occurred, unresponsive to oxygen and positive expiratory pressure therapy. Trans-oesophageal echocardiography revealed myocardial contusion and tricuspid valve rupture. Dobutamine improved left ventricular function and ejection fraction resulting in an immediate improvement in arterial oxygenation and saturation. Tricuspid injury and the diagnosis of myocardial contusion are discussed. The case highlights the importance of a non pulmonary mechanism of hypoxaemia. PMID- 8403116 TI - Anaesthetic implications of cystinosis. AB - The author presents and discusses the anaesthetic care of a 12-yr-old boy with cystinosis. Cystinosis is a recessively inherited disorder of amino acid metabolism resulting in the abnormal intracellular accumulation of cystine. Anaesthetic care may be affected by variable end-organ involvement, most notably progressive renal deterioration beginning with the development of Fanconi syndrome and progressing to overt renal failure during the first decade of life. Additional organ system involvement may lead to cirrhosis with portal hypertension, diabetes mellitus, and hypothyroidism. Identification of the extent of end-organ involvement during the preoperative evaluation will help in the provision of safe anaesthetic care for such patients. PMID- 8403117 TI - [The use of magnesium sulfate during surgery of pheochromocytoma: apropos of 2 cases]. AB - Maintenance of haemodynamic stability during anaesthesia for phaeochromocytoma resection is still a challenge. If magnesium sulfate is widely used for the control of arterial pressure during preeclampsia, its use during phaeochromocytoma resection has only been published by one author. We describe two cases where magnesium sulfate is the main agent used to control arterial pressure during resection of a phaeochromocytoma. Magnesium sulfate's hypotensive, antiarrhythmic and antiadrenergic properties are reviewed. The total doses administered were 11 g and 12 g, given as an infusion and boluses. Magnesium sulfate could be part of the anaesthetist's pharmacopoeia during phaeochromocytoma resection. PMID- 8403118 TI - Anaesthesia for cervical laminectomy in a patient with total anomalous pulmonary venous connection. AB - A 45-yr-old male patient, having total anomalous pulmonary venous connection (TAPVC) underwent cervical laminectomy under general anaesthesia without complication. The case was diagnosed preoperatively by angiocardiography. An uncomplicated patient of TAPVC can undergo non-cardiac surgery with due precaution against air-embolism and hypoxaemia. PMID- 8403119 TI - Plasma cholinesterase deficiency in a neonate. AB - We report a two-day-old infant who had a period of apnoea lasting six hours following the intravenous administration of succinylcholine (Sch). The results of her plasma cholinesterase level and dibucaine number indicate a congenital absence of plasma cholinesterase (PChE) enzyme, although both parents and siblings had normal cholinesterase levels and dibucaine numbers. This is believed to be the youngest reported case of prolonged apnoea after the administration of succinylcholine. PMID- 8403120 TI - Epidural morphine reduces the risk of postoperative myocardial ischaemia in patients with cardiac risk factors. AB - Perioperative myocardial ischaemia is a predictor of postoperative cardiac morbidity (PCM). Epidural anaesthesia and adequate perioperative analgesia have been shown to improve myocardial oxygen dynamics due to interruption of pain and sympathetic pathways. The aim of the present study was to compare the incidence of ischaemia after either general anaesthesia followed by parenteral analgesia with morphine or combined epidural/general anaesthesia followed by analgesia with epidural morphine. In a prospective observer-blinded analysis of the occurrence of ischaemia, 55 patients (epidural = 29/parenteral = 26) scheduled for elective surgery with defined risks for ischaemic cardiac disease were entered and followed for 24 hr after surgery with two-lead continuous Holter monitoring. Groups were similar with respect to age, weight, modified Goldman (Detsky) risk classification and the use of cardiac medications. Fewer patients receiving the epidural anaesthesia/analgesia had ischaemic episodes (17.2 vs 50.0%, P = 0.01), and tachyarrhythmias (20.7 vs 50.0%, P < 0.05). Epidural patients had a four-fold reduction of the relative risk for either event (P < 0.001). All ischaemic events were asymptomatic and unrecognized (silent). All major morbid events (n = 5) (MI, congestive heart failure and death) occurred in patients who had perioperative episodes of ischaemia. There were three distinct peaks in onset of ischaemia, at 1-4 hr, 9-12 hr and 22-24 hr postoperatively. One third of postoperative ischaemic events occurred within the first four hours after operation and lasted from 1 to 31 min. Forty-two percent of ischaemic episodes were associated with a heart rate > 100 bpm, or an increase of 20% over the baseline heart rate. We conclude that epidural anaesthesia/analgesia reduces but does not eliminate the risk of myocardial ischaemia and tachyarrhythmia. We were unable to determine any associated reduction in the risk of PCM. PMID- 8403121 TI - Transsartorial approach for saphenous nerve block. AB - Blockade of conduction in the saphenous nerve is important in providing surgical anaesthesia in the lower leg. Unfortunately, previously described techniques have lacked clinical effectiveness in practice. We developed a transsartorial approach for conduction block of the saphenous nerve. We first confirmed its potential clinical utility in 12 cadaveric specimens by demonstrating that the saphenous nerve was consistently stained by injections of methylene blue. Subsequently, we compared the relative rates of successful saphenous nerve block and the extent of conduction block provided by three techniques: (1) transsartorial saphenous nerve block (TSSNB), (2) above knee femoral paracondylar field block (FPFB), and (3) below knee field block (BKFB) of the saphenous nerve in 20 ASA I volunteers. The transsartorial saphenous nerve block proved to be highly successful (80% success rate) and was superior to the other two approaches in providing cutaneous analgesia to pinprick in the saphenous nerve distribution (P < 0.05). The success rates of the BKFB and FPFB were 65% and 40% respectively. A successful block with the transsartorial approach provided complete anaesthesia of the medial malleolus in 94% of subjects whilst the BKFB and FPFB provided complete anaesthesia of the medial malleolus in less than 40% of the successful blocks. We recommend the transsartorial approach for more effective block of the saphenous nerve. PMID- 8403122 TI - A modified retrobulbar block for eye surgery. AB - A modified retrobulbar block (MRB) using a single superomedial injection was compared with the classical retrobulbar block (RB) and peribulbar block (PB) in a randomized, prospective, surgeon-blinded study involving 150 patients undergoing cataract surgery. No serious complication occurred in any of the patients. The MRB produced higher rates of total akinesia in the orbicularis and all the extraocular muscles, which were statistically significant for the orbicularis, superior, inferior and lateral rectus and oblique muscles when compared with RB, and for the superior rectus and oblique muscles when compared with PB. MRB required less supplemental blocks, provided good operating conditions for the surgeon, and achieved high patient acceptance. It is concluded that MRB is a useful alternative method of ocular block for cataract surgery. PMID- 8403123 TI - A mass balance model for the Mapleson D anaesthesia breathing system. AB - A mathematical model is described which calculates the alveolar concentration of CO2(FACO2) in a patient breathing through a Mapleson D anaesthesia system. The model is derived using a series of mass balances for CO2 in the alveolar space, dead space, breathing system limb volume and reservoir. The variables included in the model are tidal volume (VT), respiratory rate, fresh gas flow rate (Vf), dead space volume, I:E ratio, and expiratory limb volume (Vl) time constant of lung expiration, and carbon dioxide production rate. The model predictions are compared with measurements made using a mechanical lung simulator in both spontaneous and controlled ventilation. Both the model and the experimental data predict that at high fresh gas flow rates and low respiratory rates, FACO2 is independent of Vf; at low fresh gas flow rates and high respiratory rates, FACO2 is independent of respiratory rate. The model and the data show that the VT influences FACO2, independent of minute ventilation alone, during both partial re breathing and non-rebreathing operation. Therefore, describing the operation in terms of minute ventilation is ambiguous. It is also shown that Vl influences FACO2 such that, for any combination of patient and breathing-system variables, there is a Vl that minimizes the Vf required to maintain FACO2. In addition, expiratory resistance can increase the fresh gas flow rate required to maintain a given FACO2. The respiratory patterns observed with spontaneous and controlled ventilation are responsible for the difference in Vf required with each mode of ventilation. PMID- 8403124 TI - Postoperative epidural analgesia. PMID- 8403125 TI - Polamedco endotracheal tubes. PMID- 8403126 TI - No more blood patch in cancer patients. PMID- 8403127 TI - Propofol and chemotherapy emesis. PMID- 8403128 TI - MAOI and cardiac surgery. PMID- 8403129 TI - Paramyotonia and MH. PMID- 8403130 TI - Absence of a capnograph trace after confirmed tracheal intubation. PMID- 8403131 TI - ARDS and sepsis--definitions and new therapy. PMID- 8403132 TI - Regional anaesthetic technique and the incidence of tourniquet pain. AB - The influence of regional anaesthetic technique on the incidence of lower extremity tourniquet pain was evaluated. We studied 60 patients undergoing orthopaedic procedures of the lower extremity with the use of a pneumatic tourniquet and anticipated inflation of 60 min or longer. Three different anaesthetic techniques were selected by random and draw; spinal anaesthesia (SAB) with plain 0.5% bupivacaine (15 mg) and 0.2 mg epinephrine added, lumbar epidural anaesthesia (EA) with 2% mepivacaine and 1:200,000 epinephrine added, and epidural anaesthesia (AEA) with the same solution alkalinized with bicarbonate. Onset and level of sensory blockade were determined by loss of painful sensation to pinprick. The incidence of tourniquet pain was determined at 15-min intervals or by patient complaint, by an observer unaware of group. Time to onset of pain, amount of treatment (i.v. fentanyl), and sensory level at the time of pain were determined. The SAB was compared with EA and AEA, and EA was compared with AEA. The SAB group was older. The sensory level achieved and duration of tourniquet inflation did not differ among groups. The incidence of tourniquet pain was lower with SAB than with EA and lower with AEA than with EA. There was no difference between SAB and AEA. This study demonstrated a lower incidence of tourniquet pain with spinal anaesthesia than with epidural anaesthesia to the same sensory level. However, this advantage is eliminated if the epidural anaesthetic was performed with an alkalinized local anaesthetic. PMID- 8403133 TI - [Elevation of arterial pressure during surgery with a pneumatic tourniquet: an independent action of renin?]. AB - The aim of this study was to evaluate the effectiveness of captopril for the prevention of the increase of arterial pressure during orthopaedic surgery requiring the application of lower limb tourniquets with balanced anaesthesia. Twenty consecutive patients were included in the study and were randomly divided into two groups. The first (n = 10) received 50 mg captopril orally together with the preanaesthetic medication, the second (n = 10) received a placebo at the same time. The different variables studied (arterial pressure, heart rate) were continuously measured. This study demonstrated that the pretreatment with captopril did not prevent an increase of the arterial pressure during the application of a tourniquet. The means of the systolic and diastolic arterial pressures at the end of the application of the tourniquet were 128/86 and 128/81 in the captopril group and the placebo group, respectively. This result shows that the renin-angiotensin system does not significantly contribute to the increase of the arterial pressure induced by a tourniquet. PMID- 8403134 TI - Postdural puncture headache: a randomized prospective comparison of the 24 gauge Sprotte and the 27 gauge Quincke needles in young patients. AB - This study was designed to compare the frequency of postdural puncture headaches (PDPH) using the 24 gauge Sprotte and the 27 gauge Quincke spinal needles in a population of patients less than 45 yr of age undergoing spinal anaesthesia for non-obstetrical surgery. Patients were randomly assigned to receive spinal anaesthesia with either the 24 gauge Sprotte spinal needle (n = 46) or the 27 gauge Quincke spinal needle (n = 47). Patients were interviewed on either postoperative day one or two and on postoperative day three. A PDPH was defined as a headache involving the occipital or frontal areas that is made worse when assuming either the sitting or standing position. Ninety-three patients were included in the analysis of data. The overall incidence of PDPH was 14% (13 of 93), and no difference was found between the Sprotte (15.2%) and Quincke (12.8%) needles. The distribution of the PDPHs by severity was not different between the two groups. None of the 13 patients with PDPHs required on epidural blood patch for relief of symptoms. Both the Sprotte needle and the Quincke needles were judged as easy to use and both required the same number of attempts in order to locate cerebrospinal fluid (first attempt successful: 73.9% versus 66%). Neither patient satisfaction nor the acceptability of spinal anaesthesia for a future procedure was adversely affected by the occurrence of a PDPH. The results of this study suggest that the risk of PDPH after spinal anaesthesia in young patients is similar using either the 24 gauge Sprotte or the 27 gauge Quincke spinal needle. PMID- 8403135 TI - Oral clonidine premedication attenuates the haemodynamic effects associated with ketamine anaesthetic induction in humans. AB - Induction of anaesthesia is often associated with undesirable variations in blood pressure and heart rate. Clonidine has been demonstrated to attenuate many of these undesirable effects when used as a premedicant. Other alpha 2 adrenergic agonists have been used to ameliorate the cardiostimulatory effects of ketamine in animals but there are few data on the use of this combination in humans. The effect of oral clonidine premedication, 5 micrograms.kg-1 on the haemodynamic changes induced by i.v. ketamine was studied in 42 patients volunteers. Ninety minutes before surgery, patients randomly received clonidine (C), diazepam (D), or a placebo (P) in a double-blinded fashion. Anaesthesia was induced with a ketamine infusion of 1 mg.kg-1 x min-1 until loss of consciousness. Heart rate and phasic blood pressure were measured noninvasively prior to induction, before and up to seven minutes after tracheal intubation. There were no differences in demographics or baseline vital signs among the three groups. With ketamine administration, increases in heart rate and blood pressure were less in those patients given C preoperatively than in those who received either D or P. The peak increase in mean blood pressure was 39% (C) versus 70% (D) and 55% (P) (P < 0.01). Heart rate increased by a maximum of 20% (C) versus 41% (D) and 46% (P) (P < 0.01). We conclude that oral clonidine attenuates the hyperdynamic effects of anaesthetic induction with i.v. ketamine. PMID- 8403136 TI - Lumbar epidural anaesthesia prevented prostaglandin E1-induced diuretic effect in enflurane anaesthetized patients. AB - Prostaglandin E1 (PGE1) is used to induce deliberate hypotension during anaesthesia. The purpose of this study was to compare the PGE1-induced diuretic effect in anaesthetized patients with and without lumbar epidural anaesthesia. The changes in haemodynamic variables, urinary flow, one-hour creatinine clearance (Ccr), and fractional excretion of sodium (FENa) during injection of PGE1 or a vehicle were compared in 42 surgical patients during enflurane anaesthesia with lumbar epidural anaesthesia (EPI group) with those in 44 surgical patients during enflurane anaesthesia alone (GA group). Patients in the GA group demonstrated increases in urinary flow (114 +/- 46%) (mean +/- SE), Ccr (74 +/- 26%), and FENa (54 +/- 23%) during PGE1 infusion, which were not observed in the patients in the EPI group. Mean arterial pressure decreased during PGE1 infusion from 92 +/- 3 to 70 +/- 2 mmHg in the GA group (P < 0.01) and from 85 +/ 2 to 65 +/- 1 mmHg in the EPI group (P < 0.01). Plasma antidiuretic hormone concentration during surgery was 12.5 +/- 2.6 U.L-1 in the GA group and 2.3 +/- 0.8 U.L-1 in the EPI group (P < 0.001). It is concluded that PGE1-induced diuresis was prevented by lumbar epidural anaesthesia. PMID- 8403137 TI - Publication of abstracts presented at anaesthesia meetings. AB - To determine the publication rate of abstracts as peer-reviewed manuscripts during the five years subsequent to their presentation, the rates of publication of abstracts that were presented at meetings of four anaesthesia societies (American Society of Anesthesiologists (ASA), International Anesthesia Research Society (IARS), Anaesthesia Research Society (ARS) and Canadian Anaesthetists' Society (CAS), in 1985 were determined. Abstracts (total = 215) from each of the four meetings were selected (ASA n = 114/573 total, IARS n = 39/119, ARS n = 33/99 and CAS n = 29/58) and their appearances in the literature as peer-reviewed manuscripts were determined using MEDLINE for the years 1985 to 1990 under the surname of the presenting author. The contents of the abstracts were compared with those of the resultant manuscripts. The proportions of abstracts from each of the four societies that were published as manuscripts were compared. We found that the mean proportion of abstracts from all four societies that were published as manuscripts within three years of presentation was 44% and within five years 50%. The proportions of abstracts that were published as manuscripts from the four societies were similar. Of the abstracts that were published as manuscripts, 13% from the ASA, 16% from the IARS, 16% from the ARS and 0% from the CAS were published four or five years after abstract presentation. Although the overall proportion of abstracts that was published within five years of presentation did not differ from the rate of publication within three years, we recommend that a uniform policy with respect to the time interval for citation of abstracts be adopted for all anaesthesia journals. PMID- 8403139 TI - Adult respiratory distress syndrome after radical neck dissection. AB - The clinical management of an unusual case of postoperative ARDS is reported. A few hours following neck surgery and septic insult, the patient developed unexpected ARDS. Aetiologic and supportive treatment were successfully instituted and after 72 hours of intensive therapy, the patient's clinical status improved. The very short time lapse between the septic insult and appearance of ARDS is emphasized. A brief literature review on aetiology, diagnosis and therapy of sepsis, as well as some pertinent aspects concerning the pathogenesis of ARDS and its linkage to sepsis are presented. PMID- 8403140 TI - Transoesophageal echocardiography and the intraoperative diagnosis of left atrial invasion by carcinoid tumour. AB - A 41-yr-old woman with pulmonary carcinoid tumour presented for thoracotomy and lung resection. However, intraoperative transoesophageal echocardiography (TEE) revealed that the tumour had invaded the left atrium, and the planned resection was aborted to allow resection under cardiopulmonary bypass at a later date. Although the incidence of cardiac involvement by lung cancer at the time of pulmonary resection is unknown, transoesophageal echocardiography can be useful in identifying the extension of hilar lung tumours. This should preferentially be done preoperatively, but can be done intraoperatively as described in this case report. PMID- 8403138 TI - Regional anaesthesia for intraocular surgery. AB - The role of anaesthetists in providing local anaesthesia for intraocular surgery has changed over the past decade. No longer confined to the interested few, more and more anaesthetists are involved in monitored care and/or are performing eye block anaesthesia. This review summarizes the information related to eye block anaesthesia. The salient features of the orbital anatomy important for safe conduct of eye block anaesthesia are described. The techniques for retrobulbar and peribulbar anaesthesia, including facial nerve blocks, anaesthetic mixture, types of needles, and softening the eye are presented. Complications such as retrobulbar haemorrhage, globe penetration/perforation, visual impairment, brainstem anaesthesia, muscle injury, and oculocardiac reflex are explored. The implications of anticoagulant therapy are examined. The choice between retrobulbar and peribulbar blocks and the role of anaesthetists are discussed. PMID- 8403141 TI - Spinal anaesthesia for caesarean section after Harrington instrumentation. AB - A case is presented of a 33-yr-old parturient with Harrington fusion of her spine who received spinal anaesthesia with 15 mg hyperbaric bupivacaine for Caesarean delivery. Multiple attempts of needle insertion in both midline and paramedian at the L3-4 interspace were unsuccessful, whereas the procedure was performed uneventfully at the midline of the L5S1 interspace. The anatomical considerations and difficulties in achieving reliable epidural anaesthesia after Harrington fusion are reviewed. Spinal anaesthesia performed at the L5S1 interspace may provide less technical difficulty and a more reliable result in such patients. PMID- 8403142 TI - Lumbar epidural anaesthesia for inguinal hernia repair in low birth weight infants. AB - In view of the complications of general, spinal, and caudal anaesthesia for inguinal hernia repair in high-risk neonates, an evaluation of lumbar epidural anaesthesia (LEA) was undertaken to assess its technical feasibility, effectiveness and incidence of complications. In 18 consecutive cases, gestational age 26 +/- 2.6 wk, birth weight 877 +/- 310 g, 16 (89%) had bronchopulmonary dysplasia and 12 (67%) were oxygen-dependent at the time of surgery. Using a standard loss of resistance technique and a 4.0 cm 20 G epidural needle, the epidural space was positively identified on the first attempt in 16 (89%), and on the second attempt in 2 patients (11%). Reflux of 0.9% saline used to identify the epidural space was blood tinged in two cases. Epidural analgesia was achieved in all cases with bupivacaine 0.25% with and without 1:200,000 epinephrine, 0.75 ml.kg-1 for the first two cases, and subsequently 1.0 ml.kg-1. In 15 patients (83%), good operating conditions were achieved with epidural analgesia alone. Inhalational anaesthesia supplementation was necessary in three cases (17%). In the first two patients, the level of analgesia (T8) was insufficient to control the response to traction on the hernial sac. In one infant, analgesic to T4, whose surgery was inadvertently delayed for four hours, inhalation anaesthesia was needed to control restlessness rather than pain. Ten infants were analgesic to T2, four to T4, two to T6 and two to T8. Intraoperative periodic breathing was seen in seven infants (39%), four with oxyhaemoglobin desaturation to 75%, and two to 85%. All responded to increased FIO2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403143 TI - Displacement of lidocaine from the lung after bolus injection of bupivacaine. AB - The purpose of this study was to determine whether lidocaine was displaced from the lung after bolus injection of bupivacaine. Fourteen anesthetized rabbits were randomly assigned to either a bupivacaine or a control group. Lidocaine was infused at a rate of 10 mg.kg-1 x hr-1. After one hour of infusion, a bolus of bupivacaine (1 mg.kg-1) in normal saline (0.2 ml.kg-1) was injected into the central venous circulation in the bupivacaine group. The control group was injected with normal saline. After bolus injection, arterial blood samples were collected serially from an internal carotid artery at 1.2-sec intervals for 24 sec. The baseline concentration of lidocaine was 3.0 +/- 0.1 micrograms.ml-1 in the bupivacaine group and 3.2 +/- 0.1 micrograms.ml-1 in the control group (NS). Arterial concentrations of lidocaine increased to a maximum of 4.7 +/- 0.2 micrograms.ml-1 in the bupivacaine group (P = 0.0001). No increases were seen in the control group. These findings indicate that lidocaine was displaced from the lung into the blood after bolus injection of bupivacaine. The amount of lidocaine displaced during the first passage of bupivacaine through the lung was calculated to be 92.3 +/- 9.7 micrograms. It is concluded that lidocaine is displaced from the lung after bolus injection of bupivacaine. PMID- 8403144 TI - Fibreoptic bronchoscopy and double-lumen endobronchial tubes. PMID- 8403145 TI - Tracheal tube kinking. PMID- 8403146 TI - Oral midazolam and post-operative behaviour in children. PMID- 8403147 TI - Emergency airway management (1) PMID- 8403148 TI - Emergency airway management (2) PMID- 8403149 TI - Accidental total spinal block (1) PMID- 8403150 TI - Accidental total spinal block (2) PMID- 8403151 TI - Accidental total spinal block (3) PMID- 8403152 TI - Anaesthesia pharmacoeconomics. PMID- 8403153 TI - Cortical blood flow response to hypercapnia during anaesthesia in Moyamoya disease. AB - Cortical blood flow (CoBF) was measured continuously by the laser-Doppler method to evaluate the effect of hypercapnia on cortical blood flow during ten surgical procedures in ten young patients (mean +/- SD 9.3 +/- 6.4 yr) with Moyamoya disease. The CoBF was 42.8 +/- 13.4 (ml.100 g-1.min-1) during normocapnia (PaCO2 = 39.0 +/- 2.4 mmHg), and 38.7 +/- 14.4 during hypercapnia (PaCO2 = 47.1 +/- 2.5 mmHg). There was a decrease in CoBF with hypercapnia (P < 0.05) so that the normal CoBF response to hypercapnia was impaired during surgery in the patients with Moyamoya disease. He concluded that patients with Moyamoya disease have a precarious cerebral circulation and hypercapnia may be detrimental to the cortical circulation. This suggests that normocapnia is preferable to hypercapnia in patients with Moyamoya disease during anaesthesia. PMID- 8403154 TI - Epidural test dose: lidocaine 100 mg, not chloroprocaine, is a symptomatic marker of i.v. injection in labouring parturients. AB - The authors studied the sensitivity (SN) and specificity (SP) of an epidural test dose containing either lidocaine 100 mg or 2-chloroprocaine 100 mg as symptomatic markers of intravascular injection in labouring parturients. In a prospective, double-blind and randomized fashion 48 unmedicated and labouring parturients were equally divided into three groups. After placement of a lumbar epidural catheter the normal saline group (NS) received 5 ml normal saline i.v., the lidocaine group (LD) received lidocaine 100 mg i.v., and the 2-chloroprocaine group (CH) received 2-chloroprocaine 100 mg i.v. All injections were given during uterine diastole. Within the next one to two minutes a blinded observer recorded the patient's perception of the presence of metallic or funny taste, dizziness, and tinnitus. We then calculated SN and SP of each symptom (alone and in combination) along with their positive (+) and negative (-) predictive value (PV). In both groups no symptom alone reached clinically acceptable levels of SN (< 87%). Only in the LD group, tinnitus+taste and dizziness+taste reached a SN of 100% with a SP of 81% and 69% respectively. While the -PV was 100% for both groups of symptoms, the +PV reached 42% for tinnitus+taste and 30% for dizziness+taste. We conclude that lidocaine 100 mg is a sensitive marker of intravascular injection in labouring parturients, and that tinnitus+taste is the most reliable indicator of intravenous injection. PMID- 8403155 TI - Intrathecal meperidine for elective caesarean section: a comparison with lidocaine. AB - The purpose of this study was to determine the efficacy of intrathecal meperidine in patients undergoing Caesarean section, and also to compare meperidine with heavy lidocaine. Fifty full-term pregnant women, ASA physical status I or II, presenting for elective Caesarean section under spinal anaesthesia were randomly divided into two groups with 25 in each, to receive either intrathecal meperidine or lidocaine. All patients received premedication with oral ranitidine, 150 mg, the night before surgery, and again two hours before surgery. Patients in the meperidine group were also given metoclopramide iv 10 mg one hour before surgery. After iv 20 ml.kg-1 Ringer's lactate, patients were given either 5% meperidine 1 mg.kg-1 or 5% heavy lidocaine 1.2 to 1.4 ml intrathecally. The sensory and motor blockades in all except two patients in each group who required sedation at the time of skin incision were adequate for surgery. None of the mothers suffered from any major side effects. The incidence of hypotension was higher in the lidocaine group than in meperidine group (P < 0.05). Pruritus and drowsiness were more common in meperidine group than in lidocaine group (P < 0.01). All the newborns in both groups cried immediately after birth and had an Apgar score > 7. The mean duration of postoperative analgesia was six hours in the meperidine group and one hour in the lidocaine group (P < 0.01). Postoperative analgesia requirement was less in the meperidine than in the lidocaine group (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403156 TI - A comparison of two doses of epidural fentanyl during caesarean section. AB - A prospective, randomized, double-blind study was performed to compare the analgesic efficacy and side effects of epidural fentanyl, 25 micrograms vs 50 micrograms, when used to supplement epidural anaesthesia for elective Caesarean section. Fifty ASA I and II patients were randomized into two groups: Group I (n = 24) received 25 micrograms and Group II (n = 26) received 50 micrograms of epidural fentanyl after the epidural test dose. No differences between the two groups were found on any measures of intraoperative pain, nausea, drowsiness, respiratory depression, hypotension, pruritus and neonatal outcome. The low levels of pain experienced by patients indicates that doses higher than 50 micrograms of epidural fentanyl are usually unnecessary for optimal analgesia. PMID- 8403157 TI - Oral midazolam premedication in children: the minimum time interval for separation from parents. AB - To determine the minimum time interval between oral midazolam (0.5 mg.kg-1) premedication and separation from parents that ensures a smooth separation, 30 children were assigned randomly to one of three groups (ten children per group). The groups differed only in the time interval between administration of midazolam and separation from their parents: 10, 20 or 30 min. Heart rate, systolic blood pressure, and sedation and anxiolysis scores were assessed before midazolam premedication (baseline), at the time of separation from parents, and during the application of a face mask at the induction of anaesthesia. We found that heart rate and systolic blood pressure changes were similar for all three groups throughout the study period. Sedation scores at the time of separation from parents and on application of the mask for all three groups were greater than baseline values. Sedation scores at separation did not differ among the three groups. Anxiolysis values did not differ from baseline values at any time for all three groups. We conclude that children may be separated from their parents as early as ten minutes after receiving oral midazolam, 0.5 mg.kg-1. PMID- 8403159 TI - Cerebral blood flow and cerebral blood flow velocity during angiotensin-induced arterial hypertension in dogs. AB - Pressure-passive perfusion beyond the upper limit of cerebral blood flow (CBF) autoregulation may be deleterious in patients with intracranial pathology. Therefore, monitoring of changes in CBF would be of clinical relevance in situations where clinical evaluation of adequate cerebral perfusion is impossible. Noninvasive monitoring of cerebral blood flow velocity using transcranial Doppler sonography (TCD) may reflect relative changes in CBF. This study correlates the effects of angiotensin-induced arterial hypertension on CBF and cerebral blood flow velocity in dogs. Heart rate (HR) was recorded using standard ECG. Catheters were placed in both femoral arteries and veins for measurements of mean arterial blood pressure (MAP), blood sampling and drug administration. A left ventricular catheter was placed for injection of microspheres. Cerebral blood flow velocity was measured in the basilar artery through a cranial window using a pulsed 8 MHz transcranial Doppler ultrasound system. CBF was measured using colour-labelled microspheres. Intracranial pressure (ICP) was measured using an epidural probe. Arterial blood gases, arterial pH and body temperature were maintained constant over time. Two baseline measures of HR, MAP, CBF, cerebral blood flow velocity and ICP were made in all dogs (n = 10) using etomidate infusion (1.5 mg.kg-1 x hr-1) and 70% N2O in O2 as background anaesthesia. Following baseline measurements, a bolus of 1.25 mg angiotensin was injected i.v. and all variables were recorded five minutes after the injection. Mean arterial blood pressure was increased by 76%. Heart rate and ICP did not change. Changes in MAP were associated with increases in cortical CBF (78%), brainstem CBF (87%) and cerebellum CBF (64%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403158 TI - Propofol in patients with cardiac disease. AB - Propofol is an intravenous anaesthetic which is chemically unrelated to other iv anaesthetics. Most anaesthetists are now becoming familiar with propofol's pharmacokinetic and pharmacodynamic properties. It has proved to be a reliable drug that can be used safely for induction and maintenance of anaesthesia for most surgical procedures and unlike other anaesthetic agents, it can especially be extended into the postoperative setting or intensive care unit for sedation. Propofol's greatest attributes are its pharmacokinetic properties which result in a rapid, clear emergence and lack of cumulative effects even after prolonged administration. Compared with other iv anaesthetics, the induction dose of propofol has a relatively higher incidence of respiratory depression, short-lived apnoea and blood pressure reduction that may occasionally be marked. Possible mechanisms for the hypotension may relate to (1) its action on peripheral vasculature (vasodilatation), (2) decreased myocardial contractility, (3) resetting of the baroreflex activity and (4) inhibition of the sympathetic nervous system outflow. In vitro studies indicate that propofol depresses the immunological reaction to bacterial challenge as well as the chemotactic activity. Clinical studies, in cardiac surgery, have demonstrated that propofol, in association with an opioid, is a logical anaesthetic choice. Propofol is about to receive approval for continuous iv sedation. Comparative studies of propofol and midazolam have clearly demonstrated the superiority of propofol in terms of rapid recovery and precise control of the level of sedation. PMID- 8403160 TI - Action of propofol on central sympathetic mechanisms controlling blood pressure. AB - This study was done using Wistar rats to determine if the actions of propofol (22 +/- 1, 40 +/- 2, 64 +/- 3 and 103 +/- 3 mg.kg-1 x hr-1) decreased blood pressure and heart rate through depression of brain stem vasomotor centres. All rats were given atropine to block vagal influences on the heart. Propofol decreased renal nerve activity as well as blood pressure and heart rate in a dose-dependent manner. Infusion of the lowest dose of propofol (22 +/- 1 mg.kg-1 x hr-1) had no effect on blood pressure, heart rate and renal nerve activity. Infusion of propofol at 40 +/- 2 mg.kg-1 x hr-1 decreased renal activity by 22 +/- 4% (mean +/- SEM) and at 64 +/- 3 mg.kg-1 x hr-1 it decreased renal nerve activity by 36 +/- 6%. Finally, infusion of the largest dose of propofol (102 +/- 3 mg.kg-1 x hr 1) decreased nerve activity by 50 +/- 5%. The haemodynamic changes observed in our experiments during the infusion propofol paralleled the changes in sympathetic firing, suggesting that hypotension was caused by central actions of propofol to depress sympathetic firing. In experiments with bolus injections of propofol, the renal nerve activity returned to normal before arterial pressure and heart rate recovered. Because decreases in blood pressure and heart rate were longer-lasting than changes in renal nerve activity, a part of the vasodepression and bradycardia caused by propofol likely resulted from direct actions on blood vessels and the heart. Sympathetic and cardiovascular responses to blocking neurons in the ventrolateral medulla with microinjection of glycine were depressed by propofol.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403161 TI - The influence of intravenous anaesthetics on polymorphonuclear leukocyte function. AB - Polymorphonuclear leukocytes (PMNL) play a vital role in the defence against invading bacteria. It is known that some anaesthetics inhibit PMNL function and, thus, possibly enhance perioperative infection. We investigated the effect of methohexitone, flunitrazepam, and droperidol on three bactericidal PMNL functions, i.e., superoxide anion production, hydrogen peroxide generation, and activity of released myeloperoxidase, in vitro. Approved photometrical assays were used. Superoxide anion was measured by the reduction of cytochrome C, hydrogen peroxide by the horse radish peroxidase catalysed oxidation of phenol red, and myeloperoxidase by the turnover of 2,2'-azino-di(3-ethylbenzthiazoline) sulfonic acid. Methohexitone (P < or = 0.001) and flunitrazepam (P < or = 0.01) inhibited superoxide anion production, and methohexitone (P < or = 0.01) reduced hydrogen peroxide generation but only at concentrations beyond clinical relevance. Droperidol did not cause any alteration of the PMNL functions tested. Consequently, it seems unlikely that the usual doses of methohexitone, flunitrazepam, or droperidol promote bacterial infections in vivo by impairing the activity of myeloperoxidase or by inhibiting the generation of superoxide anion or hydrogen peroxide. PMID- 8403162 TI - Vasodilators during cerebral aneurysm surgery. AB - The objective of this review is to review the anaesthetic implications of vasoactive compounds particularly with regard to the cerebral circulation and their clinical importance for the practicing anaesthetist. Material was selected on the basis of validity and application to clinical practice and animal studies were selected only if human studies were lacking. Hypotensive drugs have been used to induce hypotension and in the treatment of intraoperative hypertension during cerebral aneurysm surgery. After subarachnoid haemorrhage, cerebral blood flow is reduced and cerebral vasoreactivity is disturbed which may lead to brain ischaemia. Also, cerebral arterial vasospasm decreases cerebral blood flow, and may lead to delayed ischaemic brain damage which is a major problem after subarachnoid haemorrhage. Recently, the use of induced hypotension has decreased although it is still useful in patients with intraoperative aneurysm rupture, giant cerebral aneurysm, fragile aneurysms and multiple cerebral aneurysms. In this review, a variety of vasodilating agents, prostaglandin E1, sodium nitroprusside, nitroglycerin, trimetaphan, adenosine, calcium antagonists, and inhalational anaesthetics, are discussed for their clinical usefulness. Sodium nitroprusside, nitroglycerin and isoflurane are the drugs of choice for induced hypotension. Prostaglandin E1, nicardipine and nitroglycerin have the advantage that they do not alter carbon dioxide reactivity. Local cerebral blood flow is increased with nitroglycerin, decreased with trimetaphan and unchanged with prostaglandin E1. Intraoperative hypertension is a dangerous complication occurring during cerebral aneurysm surgery, but its treatment in association with subarachnoid haemorrhage is complicated in cases of cerebral arterial vasospasm because fluctuations in cerebral blood flow may be exacerbated. Hypertension should be treated immediately to reduce the risk of rebleeding and intraoperative aneurysmal rupture and the choice of drugs is discussed. Although the use of induced hypotension has declined, the control of arterial blood pressure with vasoactive drugs to reduce the risk of intraoperative cerebral aneurysm rupture is a useful technique. Intraoperative hypertension should be treated immediately but the cerebral vascular effects of each vasodilator should be understood before their use as hypotensive agents. PMID- 8403163 TI - Intraoperative reinfusion of whole blood using a new autoinfusion device. AB - Aortic aneurysm resection is frequently associated with considerable blood loss and requires transfusion. To minimize complications and cost many institutions use a "cell saving" method that allows reinfusion of the washed red cell fraction of blood suctioned from the operative field. The disadvantages of this technique are that homologous transfusion is regularly required to replace platelets and coagulation factors. Red cell transfusion may also be required when there is rapid major blood loss as the wash cycle may be too long to subject a patient, in a high-risk group for coronary artery disease, to anaemia. A new autoinfusion device anticoagulates blood as it is suctioned from the operative field then filters, defoams, and returns it whole to the patient without a processing time lapse. We successfully used the device in a patient for aortic aneurysm resection to reinfuse two-thirds of his blood volume shed over 80 min. Neither banked red cells nor plasma were used. His haematocrit and coagulation profile remained stable throughout surgery and recovery. The potential complications and cost of homologous transfusion were avoided. PMID- 8403164 TI - Epidural fentanyl and C-section. PMID- 8403165 TI - Regurgitation through a laryngeal mask. PMID- 8403166 TI - Tracheal intubation and cervical injury. PMID- 8403167 TI - Difficult laryngoscopy--"BURP". PMID- 8403168 TI - Equipotent dose regimens required when comparing epidural opioids. PMID- 8403169 TI - LMA for failed intubation. PMID- 8403170 TI - LMA for failed intubation. PMID- 8403171 TI - Propofol-related convulsions. PMID- 8403172 TI - Thoracic epidural anaesthesia in children--problems with retrospective data. PMID- 8403173 TI - Quality: a job well done! PMID- 8403174 TI - Patient response to laryngeal mask insertion after induction of anaesthesia with propofol or thiopentone. AB - The response to insertion of the laryngeal mask airway (LMA) following either propofol 2.5 mg.kg-1 or thiopentone 5 mg.kg-1 was assessed in two groups of patients. The purpose of the study was to ascertain which of these two induction agents provided the better conditions for insertion of the LMA. Anaesthesia was induced by propofol in 35 patients and by thiopentone in 37. Following induction, ventilation was assisted for two minutes using 50% oxygen and nitrous oxide and 2% isoflurane, before insertion of the LMA. The presence of gagging, coughing, laryngospasm and movement was noted and graded. Thiopentone was associated with an adverse response in 76% of patients, compared with propofol in 26% (P < 0.01). Gagging, laryngospasm and head movement were more common using thiopentone (P < 0.01, P < 0.05 and P < 0.05 respectively) and in 11% (P < 0.05) of the thiopentone group insertion of the LMA was impossible due to inadequate relaxation. We conclude that, using these doses, propofol is superior to thiopentone as an induction agent for insertion of the laryngeal mask airway. PMID- 8403175 TI - A comparison of regularly dosed oral morphine and on-demand intramuscular morphine in the treatment of postsurgical pain. AB - A randomized, placebo-controlled, double-blind clinical trial was conducted to compare the use of regularly dosed po morphine and on-demand in morphine in 47 patients undergoing total hip arthroplasty. Patients were randomized to receive either 20 mg (initial dose) of regularly dosed morphine (every four hours po) plus breakthrough pain medication on-demand consisting of both 10 mg morphine po and placebo im, or an equivalent regularly dosed oral placebo (every four hours) with breakthrough pain medication consisting of oral placebo and 5-10 mg morphine im. Subsequent to each request for breakthrough pain medication, the next regularly dosed oral solution was increased by 5 mg (or equivalent volume of placebo) to a maximum of 40 mg po Q4H. Time-averaged pain scores were lower on both postoperative day 1 and 2 in the group receiving regularly dosed morphine po (P < 0.05). Fewer patients requested breakthrough pain medication on both days in the oral morphine group. The incidence of nausea and vomiting, and of decreased respiratory rates were similar in both groups. Regularly dosed oral morphine is inexpensive and should be compared to other methods of opioid delivery. PMID- 8403176 TI - Morphine does not affect the awakening concentration of sevoflurane. AB - This study was designed to determine whether morphine 0.1 mg.kg-1 i.v. given intraoperatively altered the end-tidal concentration of sevoflurane which is associated with eye opening to verbal command. We studied 24 healthy ASA physical status I patients to determine whether morphine, or placebo administered about 60 min before the end of surgery affected recovery from sevoflurane/oxygen anaesthesia. During anaesthesia no other anaesthetics or drugs were given. After surgery, end-tidal sevoflurane concentration was reduced gradually at the rate of less than 0.01%.min-1. The end-tidal concentration at the time patients could respond to verbal command was recorded as MACawake. The MACawake was 0.58 +/- 0.12% (mean +/- SD) for the control group to whom placebo had been administered, and 0.57 +/- 0.11% for morphine group to whom morphine had been administered. In both groups, the MACawake decreased with age, and the ratio to age-adjusted sevoflurane MAC was 0.31 +/- 0.04 (mean +/- SD) for the control group and 0.30 +/ 0.04 for the morphine group. The ratio had no correlation with age. It is concluded that the awakening concentration of sevoflurane during recovery from anaesthesia is not affected by analgesic doses of morphine 0.1 mg.kg-1 i.v. administered intraoperatively. PMID- 8403177 TI - Premedication with sublingual triazolam compared with oral diazepam. AB - The clinical effects of a new administration form of triazolam, 0.2 mg sublingual (sl) tablet, were compared with those of a 10 mg tablet of diazepam in a double blind study, in 100 ASA I-II patients scheduled for ophthalmic surgery under local anaesthesia. The sedative and anxiolytic effects of the study drugs were followed at 15 min intervals by plotting the patient's assessments on a visual analogue scale and by a trained observer. The onset of sedative and anxiolytic effect was similar. At 75 min after premedication and after the operation triazolam 0.2 mg caused deeper sedation than diazepam 10 mg according to the observer (P < 0.001, P < 0.01) and according to the patient (P < 0.01, P < 0.05). Ten patients (20%) in the triazolam group and one in the diazepam group were assessed to be too sedated during the operation. All these patients were 61-70 yr old. The study drugs resulted in equal reduction of anxiety during the preoperative period. Both premedications provided good patient comfort but caused total amnesia only to one patient in each group. Neither caused any severe cardiorespiratory or other side effects. It is concluded that 0.2 mg triazolam sl produces deeper sedation than 10 mg oral diazepam. For elderly patients this dose is excessive for intraocular surgery. PMID- 8403178 TI - Improved peribulbar anaesthesia with alkalinization and hyaluronidase. AB - A prospective double-blind randomized study was carried out to determine the effect of pH and the addition of hyaluronidase to a mixture of lidocaine and bupivacaine on the efficacy of peribulbar anaesthesia. One hundred patients were assigned to one of five groups. All groups received a solution of two parts bupivacaine (0.75%) and one part lidocaine (2%) (with 1:100,000 adrenaline) as the base components of their anaesthesia. Group 1 received only the bupivacaine lidocaine mixture, pH 3.9. Group 2 received a solution supplemented with hyaluronidase (ten units.ml-1), pH of 5.1. Group 3 received the bupivacaine lidocaine mixture alkalinized with sodium bicarbonate to a pH of 5.1, the same as solution 2. Group 4 received the mixture with hyaluronidase alkalinized to pH of 6.7. Group 5 received the bupivacaine-lidocaine mixture alkalinized to a pH of 6.7. Efficacy of each block was graded according to the degree of residual movement 30 min following injection, as described by House et al. The solution containing hyaluronidase and pH adjusted to 6.7 was found to be the most effective (P < 0.025). The presence of hyaluronidase without alkalinization did not improve the efficacy of the mixture; and similarly, alkalinization in the absence of hyaluronidase was ineffective. These results reflected the pH- and temperature-dependent thermodynamic properties of local anaesthetics, and the pH dependent activity of hyaluronidase. PMID- 8403179 TI - Current models of "quality"--an introduction for anaesthetists. AB - The purpose of this review is to provide the practicing anaesthetist with an historical perspective of quality, a summary of current models, and an introduction to the expectations of accreditors. Articles were obtained from an electronic literature search on Silver Platter using the search terms Quality, Quality assurance, Anes, and Anaes. In addition, textbooks on quality assurance in health care, quality improvement texts from business management, and accreditation documents were reviewed. Quality systems in health care are derived from business or industrial models. Study of this field is hampered by poorly defined terminology and jargon. Over the years, many different models have been used in health care, but recent studies have investigated the effectiveness of methods such as Quality Improvement. Many of the systems used by hospitals appear to have been prompted by requirements of accreditation standards. Recently, systems of hospital organization have appeared which link Quality Assurance, Quality Improvement, risk management and utilization management. Despite the confusion created by ill-defined terminology and rapid change in some definitions, anaesthetists need to be aware of the basic models of accreditation requirements. PMID- 8403180 TI - Delayed seizures following sedation with propofol. AB - Seizures occurred in two adolescents approximately six hours after sedation with propofol for bone marrow biopsy. Case #1 was a patient with chronic renal failure, hypertension, and anaemia. Case #2 had just been diagnosed with acute lymphocytic leukaemia. Neither child had experienced seizures before, and both recovered without neurological sequelae. Although other factors may have caused the seizures, the episodes have raised concerns about the safety of propofol for patients travelling home after out-patient surgery. Further study is required to explain the cause of this complication or, at least, to identify risk factors. PMID- 8403181 TI - [Glycine poisoning after percutaneous kidney surgery]. AB - We describe two cases of altered consciousness associated with nausea and vomiting after renal surgery. These events occurred respectively one and two hours after surgery. They are believed to be caused by the transfer of large amounts of irrigating solution containing glycine 1.5%. The effects of hyponatraemia (132 and 129 mmol.L-1) and glycinaemia (3518 and 8589 mmol.L-1) are discussed. We conclude on the importance of an early clinical diagnosis completed by biological investigation to allow an adequate symptomatic management. PMID- 8403182 TI - Caesarean section and phaeochromocytoma resection in a patient with Von Hippel Lindau disease. AB - This report describes the anaesthetic management of a women with a term gestation, Von Hippel Lindau disease (VHLD), and a phaeochromocytoma, scheduled for a combined phaeochromocytoma resection and Caesarean section. Von Hippel Lindau disease is characterized by diffuse haemangioblastomas of the central nervous system (CNS) and viscera. It is also associated with phaeochromocytomas and renal cell carcinomas. Patients frequently have asymptomatic spinal cord and intracranial pathology. The patient and her fetus presented a challenge because of the anaesthetic restrictions imposed by VHLD, and her pregnancy. She was also at risk of developing malignant hypertension from the phaeochromocytoma. The patient was not a candidate for regional anaesthesia because of the possibility of spinal cord haemangioblastomas. She had received adrenergic blockade with phentolamine (total 30 mg a day) and propranolol (total 40 mg a day) since the 27th wk of gestation in order to control hypertension secondary to the phaeochromocytoma. General anaesthesia was administered with aggressive management of hypertension with adrenergic blockers (labetalol 1.0 mg.kg-1 and esmolol 0.75 mg.kg-1) and sodium nitroprusside 1.5 micrograms.kg-1 (total). Before delivery of the baby, opioids, which could have resulted in a fetus with CNS depression, were avoided. After delivery, opioids (sufentanil 0.4 microgram.kg-1 x hr-1) were used to limit the use of inhalational anaesthesia which may contribute to uterine atony. Postoperative pain was managed with an intravenous narcotic infusion. Both patients had uneventful postoperative courses. PMID- 8403183 TI - Jet ventilation in a case of tracheal obstruction secondary to a retrosternal goitre. AB - Intermittent jet ventilation was used during anaesthesia in a 66-yr-old woman who had severe tracheal narrowing secondary to compression by a retrosternal goitre. The trachea was intubated by a small-bore tube, which was placed above the site of narrowing. An injector was connected to the proximal end of the tracheal tube on one side and to the anaesthesia circuit on the other. Intermittent jets of 66% nitrous oxide in oxygen via the injector resulted in adequate oxygenation and carbon dioxide elimination. Arterial blood gas analysis during jet ventilation showed PaO2 150 mmHg, PaCO2 35 mmHg and pH 7.4. It is concluded that low frequency jet ventilation may provide adequate oxygenation and carbon dioxide elimination in the presence of tracheal narrowing. PMID- 8403184 TI - Thoracic epidural anaesthesia in infants and children. AB - When compared with conventional analgesic techniques, epidural anaesthesia not only provides improved analgesia, but also has several beneficial effects on the postoperative respiratory, cardiovascular, and metabolic status of the patient. Although the efficacy and safety of caudal and lumbar epidural anaesthesia in children has been demonstrated, there is little information concerning the use of thoracic epidural anaesthesia. The purpose of our review was to evaluate the safety of thoracic epidural anaesthesia in infants and children. We retrospectively reviewed our three-year experience with thoracic epidural anaesthesia for postoperative analgesia in children. Epidural catheters were placed at the thoracic level without difficulty in 63 children ranging in age from three months to 18 yr and in weight from 3.2 to 78 kg. Postoperative analgesia was provided by the continuous infusion of a bupivacaine/fentanyl mixture, supplemented with intermittent epidural fentanyl by bolus as needed. Epidural catheters were successfully placed in all patients. No inadvertent dural punctures were noted. No episodes of respiratory depression related to epidural analgesia occurred. Minor adverse effects including pruritus occurred in six patients, three of whom required pharmacological intervention with diphenhydramine. Our review suggests that this is a safe and effective method of postoperative analgesia following thoracic surgery in children. PMID- 8403185 TI - [The effect of halothane on blood gases and arterial acid-base equilibrium in intact rats and in chemo-denervated rats]. AB - Halothane decreases the ventilatory response to hypoxia and the activity of peripheral arterial chemoreceptors, resulting in "chemical chemodenervation." In order to evaluate the role of this halothane-induced "chemical denervation" in acid-base and arterial blood gas changes, these values were measured in intact and chemodenervated rats, awake and under anaesthesia. Since the depth of anaesthesia could be modified by the anatomical chemodenervation, the ED50 of inspired halothane was determined in six rats before and after anatomical chemodenervation. To prevent haemodynamic changes due to halothane and/or anatomical chemodenervation from interfering with the results, systemic arterial blood pressure and heart rate were measured in six intact rats, awake and then anaesthetized, and in the same rats after chemodenervation, awake and then anaesthetized. In nine intact rats and in 19 chemodenervated rats, arterial pH, arterial bicarbonate concentration, and arterial blood gases (PaO2 and PaCO2) were measured before and after administration of halothane. Anatomical chemodenervation modified neither the inspired ED50 (1.1%), nor the mean arterial blood pressure or heart rate. The haemodynamic effects of halothane were comparable in intact and in chemodenervated rats. Changes in arterial blood gases and acid-base balance due to halothane in intact rats and due to chemodenervation in awake rats were not different, but there was a decrease in PaO2 and pHa, and an increase in PaCO2. In chemodenervated rats, halothane caused a further decrease in PaO2 and a further increase in PaCO2. The fact that halothane and anatomical chemodenervation have similar effects on arterial blood gases and acid base balance favours a "chemical chemodenervating" action of halothane. However, the additional effects of halothane in the anatomically chemodenervated animal show that the action of halothane on blood gases and acid-base balance is the result of multiple sites of impact on the respiratory system. PMID- 8403186 TI - Informed consent for clinical anaesthesia research. AB - Most surgical patients are first seen by an anaesthetist after admission to hospital, either the evening before or on the day of surgery. Some medical ethicists believe that an approach by an anaesthesia researcher made after admission is unethical because the hospital itself is a coercive environment, and patients have insufficient time for reflection or consultation. Others believe that an approach prior to admission may be an invasion of the patient's privacy and confidentiality. The implications of these views for anaesthesia researchers may not be apparent to research ethics boards (REBs). To determine current practice, a questionnaire concerning the membership and function of REBs and the time of obtaining informed consent was sent to each research representative of the 16 Canadian university departments of anaesthesia. Membership of REBS was similar, but not identical, in all centres. Most representation was from medical disciplines. Consent was generally obtained following the patient's admission to hospital. In one centre, the REB always requested informed consent to be obtained before the patient's admission to the hospital. Surgeons had no involvement with consent for anaesthesia research in 14 centres while in the other two they gave permission for their patients to be studied and informed patients of the potential approach by anaesthesia researchers. We conclude that it is ethically acceptable to obtain informed consent for most low-risk clinical anaesthesia research after the patient's admission to hospital. PMID- 8403187 TI - PCA in burn injuries: the subcutaneous route. PMID- 8403188 TI - Laryngeal mask airway: defining the limits. PMID- 8403189 TI - The laryngeal mask airway and ocular injury. PMID- 8403190 TI - Perforation of the trachea by a central venous catheter guidewire. PMID- 8403191 TI - Dental restoration using oral ketamine. PMID- 8403192 TI - Direct enzymic detection of endogenous oxidative base damage in human lymphocyte DNA. AB - The endogenous production of oxidative damage in DNA by free radicals released as a by-product of respiration is a likely cause of mutations which, if they occur in appropriate genes, may lead to cancer. Using an endonuclease specific for oxidized pyrimidines, in conjunction with the highly sensitive method of single cell gel electrophoresis, we have detected significant oxidative damage in untreated, freshly isolated lymphocytes from normal, healthy individuals. PMID- 8403193 TI - Cancer chemoprevention: principles and prospects. PMID- 8403194 TI - Inhibition by gonadectomy of effects of 2-acetylaminofluorene in livers of male, but not female rats. AB - The influence of gonadectomy on the effects of 2-acetylaminofluorene (AAF) in the livers of rats was studied. Groups of male and female F344 rats at 9 weeks of age were given AAF by daily gavage 5 days per week for 4 or 8 weeks for total cumulative doses of 1.0 or 2.0 mmol/kg body wt. AAF was administered either with no pretreatment or beginning 4 weeks after gonadectomy, which was performed at 5 weeks of age. In male rats AAF induced a large number of placental glutathione S transferase foci in livers by 8 weeks, while in female rats the number was about 10% of that in males. Orchidectomy decreased the AAF induction of foci in male rats by 60%, whereas ovariectomy had no effect in female rats. Similarly, orchidectomy decreased DNA adduct levels approximately 85% in male rats given AAF by gavage for 4 weeks. In ovariectomized female rats at 4 and 8 weeks hepatic DNA adduct levels were somewhat elevated (< 50%) as compared to intact controls. The zone of glutamine synthetase-positive hepatocytes around the central vein was reduced by AAF exposure of male, but not female, rats. Male rats displayed a larger zone than females and the zone in males was reduced to the level of females by orchidectomy. Orchidectomy also diminished the effect of AAF on glutamine synthetase-positive cells. Thus, the induction of neoplastic conversion by AAF in rat liver, the extent of DNA adduct formation and the reduction of the glutamine synthetase-positive zone of hepatocytes were greater in males than females and were dependent upon the hormonal status of males. PMID- 8403195 TI - Cytochrome P450 forms in the rodent lung involved in the metabolic activation of food-derived heterocyclic amines. AB - The metabolic activation of the promutagens 2-amino-3,8- dimethylimidazo[4,5 f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) by rat and mouse lung microsomes was studied using Salmonella mutagenicity (strain TA98). Lungs from uninduced animals were found to activate all three compounds. A 4-6 fold higher mutagenic activity was obtained with IQ compared to MeIQx and the mutagenic response of PhIP was 1-2 orders of magnitude lower than that of IQ. In order to characterize the forms of P450 in the lung responsible for the metabolic activation of these food mutagens Western blots were performed with microsomes and partially purified P450 fractions from the lung. Western blots revealed the presence of cytochrome P450 2A, 2B and 4A forms in untreated rats. In the lung CYP 1A1 was only detectable after BNF treatment of rats. The CYP 4A isozymes, which have not previously been described in the rat lung, were further identified after PCR amplification from lung mRNA as 4A2 and 4A8. Antibody inhibition studies showed that CYP 2A3 catalyzed a major part (70%) of the metabolic activation of IQ by uninduced rat lung microsomes. The metabolic activation of MeIQx was not influenced by this antibody. An antibody against CYP 2B isozymes also partially inhibited the activation of IQ by uninduced rat lung microsomes. However, since induction of CYP 2B isozymes in the liver by phenobarbital treatment did not increase the metabolic activation of the heterocyclic amines over controls it is unlikely that the rat lung CYP 2B1 is participating in the activation of heterocyclic amines. The inhibition of the IQ-dependent mutagenicity by the CYP 2B antibody is probably due to cross-reaction with CYP 2A3. Alfa-naphthoflavone (ANF), considered to be a specific inhibitor of CYP 1A isozymes at 10 microM, partly inhibited the activation of IQ (30-40%) and MeIQx (60-80%) by uninduced rat and mouse lung microsomes. Upon pretreatment of rats with BNF, lung microsomes activated MeIQx at a rate that was 2-10-fold higher than control lung microsomes, whereas the increase in EROD activity was approximately 100-fold in the same lung preparations. These results suggest that CYP 1A1 may not be the enzyme responsible for the activation of MeIQx in the control rat despite the inhibition with ANF. It is likely that ANF can inhibit other P450 enzymes in the lung, including CYP 2A3.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8403196 TI - The sensitivity to DNA topoisomerase inhibitors in L5178Y lymphoma strains is not related to a primary defect of DNA topoisomerases. AB - DNA topoisomerase-targeting antitumor drugs are potent inducers of protein concealed strand breaks in mammalian cells and act by trapping DNA topoisomerases on chromosomal DNA in the form of drug-enzyme-DNA cleavable complexes. It has been proposed that the cleavable complex is an unusual form of DNA damage that elicits cellular responses analogous to those caused by DNA damaging agents. The relationship between topoisomerase-targeting drug-induced damage and radiation induced damage has been investigated by analyzing the properties of DNA topoisomerases in mouse L5178Y lymphoma strains that are cross-sensitive to topoisomerase I-II inhibitors and to UV light or X-ray irradiation. The strains are LY-R, isolated from L5178Y cells on the basis of increased resistance to ionizing radiation, and strain LY-S, isolated from LY-R cells following a spontaneous increase in the sensitivity to ionizing radiation. LY-S cells, deficient in the rejoining of DNA double-strand breaks, show enhanced sensitivity to topoisomerase II-targeting inhibitors, whereas LY-R cells have an increased sensitivity to UV radiation and to the topoisomerase I inhibitor, camptothecin. The cellular availability of DNA topoisomerase I and II and the sensitivity of the enzymes to their specific inhibitors have been measured in the two related strains. In the LY-R strain, we found a 30% decrease in topoisomerase I content but no difference in camptothecin sensitivity, while no quantitative or qualitative differences were observed for the topoisomerase II. The results indicate that variations in sensitivity of the L5178Y strains to topoisomerase inhibitors are unlikely to be related to primary defects of the target enzymes, and thus it is possible that common pathways exist for processing of topoisomerase- and radiation-induced damage. PMID- 8403197 TI - Reduction of cell proliferative activities of gastric stump adenomatous hyperplasias after bile reflux diversion in rats. AB - Previously we reported the majority of lesions induced by bile reflux, in the absence of chemical carcinogens, in the rat remnant stomach to consist primarily of gastric type and secondarily of intestinal type cells, and that they are reversible after diversion of bile reflux. The present study was designed to evaluate changes in proliferative activities in cells of each type under these conditions. The frequency of adenomatous hyperplasia (AH) induced in the gastric stump mucosa by duodenal content reflux after Billroth II partial gastrectomy (BII) increased until the 54th week of the experiment. Roux-en-Y (RY) surgical procedure which prevents duodenal reflux performed at the 24th or 36th week after BII led to a decrease in AH. Cell content of the lesions was analyzed using routine H&E staining, immunohistochemical staining for pepsinogen isoenzyme 1 and histochemical procedures for mucins (paradoxical concanavalin A, galactose oxidase Schiff and sialidase galactose oxidase Schiff reactions) and proliferation in each compartment evaluated by an immunohistochemical method using bromodeoxyuridine (BrdU) and a monoclonal antibody against BrdU. At the 54th week the number of BrdU-labeled cells per normal pyloric column was significantly (P < 0.05) increased to 10.63/pit after the BII operation, while it diminished to 5.23/pit after RY diversion, this being the same level as with the RY procedure alone. AH maintained a high rate of BrdU incorporation at 12.7% after BII operation, which was also significantly reduced (P < 0.01) to 7.0% by the RY surgery. The intestinal type cell showed highest (22.2%), the surface mucous type cell showed the next (16.5%) and the pyloric gland type cell showed lowest (5.2%) BrdU labeling indices after BII operation. All the cell types in AH showed similar proportional decreases in BrdU incorporation after RY diversion. Thus surgical intervention reverses the cell proliferation caused by bile reflux in the gastric stump. PMID- 8403198 TI - Studies on liver tumor promotion in the rat by orotic acid: dose and minimum exposure time required for dietary orotic acid to promote hepatocarcinogenesis. AB - Our earlier studies indicated that orotic acid, a precursor for pyrimidine nucleotide biosynthesis, exerts a promoting effect on rat hepatocarcinogenesis. The present study was designed to determine the optimum conditions of exposure to orotic acid required for promotion of hepatocarcinogenesis in the initiated rats. The first series of experiments was designed to determine the optimum dose of orotic acid needed to exert its liver tumor promoting effect. Accordingly male Fischer rats were given diethylnitrosamine (200 mg/kg, i.p.) or 0.9% NaCl. One week later carcinogen-injected rats were divided into six groups and fed either basal diet or the same diet containing 0.1, 0.5, 1, 2 or 4% orotic acid. Rats given 0.9% NaCl were fed 4% orotic acid. Two-thirds partial hepatectomy was performed on all animals 10 weeks after starting on their respective diets, and all groups were killed 3 weeks thereafter. Analysis of gamma-glutamyltransferase positive foci and nodules revealed that 0.5-1% orotic acid in the diet is sufficient to exert a significant promoting effect on the selective growth of initiated hepatocytes, while higher concentrations of orotic acid were only marginally more effective. No gamma-glutamyltransferase-positive foci were observed in animals given 4% orotic acid diet following saline injection. Using 1% orotic acid as the promoting regimen, in the next series, the minimum exposure time required for dietary orotic acid to promote liver carcinogenesis was determined. Male Fischer 344 rats were given i.p. either 1,2-dimethylhydrazine dihydrochloride (100 mg/kg) or 0.9% NaCl 18 h after 2/3 partial hepatectomy. After 1 week of recovery one group of rats was continued on a semisynthetic basal diet, while others were transferred to the same basal diet containing 1% orotic acid. Rats that were on the 1% orotic acid diet were progressively transferred to the basal diet after 5, 10, 20, 29 and 40 weeks of exposure. All rats were sacrificed 54 weeks after the beginning of the experiment. The results indicate that 100% of the initiated rats developed hepatic nodules whether or not they were exposed to an orotic acid-containing diet. However, the incidence of hepatocellular carcinoma was greatly increased in animals exposed to the orotic acid diet, with 42% incidence in initiated rats given orotic acid diet for 10 weeks and up to 75% in those exposed to this diet for 40 weeks. Further, promotion by orotic acid exhibited a high metastatic potential with 33-60% metastasis to the lungs.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8403199 TI - K-ras mutations in aberrant crypt foci, adenomas and adenocarcinomas during azoxymethane-induced colon carcinogenesis. AB - Ras mutations are an important early event in a number of carcinogen-induced rodent tumors. Colon carcinogenesis induced in rats by azoxymethane is a useful model as it mimics the adenoma-carcinoma sequence observed in humans. In addition, aberrant crypt foci develop in the rat and these lesions appear to be potentially important precursors to adenomas in colorectal cancer. Recent studies have shown that specific K-ras codon 12 and 13 mutations are present in up to 66% of carcinogen-induced rat colon adenocarcinomas. We studied the frequency of these mutations during the aberrant crypt focus-adenoma-carcinoma sequence in azoxymethane-induced Fisher F344 rats. K-ras codon 12 GAT and codon 13 GAC mutations were detected with a sensitive assay based on the amplification of DNA using the polymerase chain reaction. No mutations were present in normal mucosa. Of 27 aberrant crypt foci, K-ras mutations were identified in 2 lesions containing 5 and 10 aberrant crypts, respectively. Mutations were present in 1 of 23 and 10 of 27 adenomas and adenocarcinomas, respectively. These data suggest that K-ras mutations play a role during the stages of carcinogenesis in azoxymethane-induced rat colon cancer. The demonstration of a genetic mutation in aberrant crypt foci provides further evidence for the significance of these lesions as precursor markers of malignant potential during colorectal tumorigenesis. PMID- 8403200 TI - Cytochrome P450-dependent metabolism and mutagenicity of 15,16-dihydro-11 methylcyclopenta[a]phenanthren-17-one and their implications in its carcinogenicity. AB - Methylation of the non-carcinogen 15,16-dihydrocyclopenta[a]phenanthren-17-one (CPP-17-one) at the bay region to form 11-CH3-CPP-17-one confers carcinogenic potential. In the present study we have investigated the in vitro metabolism and mutagenicity of the methylated compound by hepatic microsomal preparations from rats pretreated with various prototype inducers of cytochrome P450 proteins in order to provide a rationale for this marked difference in carcinogenic activity. The most effective metabolism of 11-CH3-CPP-17-one occurred in the presence of Aroclor 1254-induced microsomes, the principal metabolites being oxidative products of the A- and D-rings and of the methyl substituent. When benzo[a]pyrene induced microsomes served as the metabolising system, the major A-ring metabolite was the 3,4-diol. A similar metabolic pattern was seen with microsomes from rats treated with 11-CH3-CPP-one itself, but the overall effect of metabolism was lower than that observed with benzo[a]pyrene-treated microsomes but higher than that of control animals. In contrast, microsomes from rats treated with clofibrate, dexamethasone, isoniazid and phenobarbitone failed to enhance the metabolism of 11-CH3-CPP-17-one when compared with control microsomes and the metabolites reflected primarily oxidation of the D-ring. When 11-CH3-CPP-17-one was employed as a promutagen in the Ames test, a mutagenic response was evident only in the presence of microsomes from benzo[a]pyrene-induced rats, but induction with phenobarbitone, isoniazid, dexamethasone, clofibrate and the compound itself, failed to elicit a positive mutagenic response. When 3,4 dihydroxy-11-CH3-CPP-17-one served as the promutagen, a mutagenic response was observed in the presence of benzo[a]pyrene-induced and, to a lesser extent, 11 CH3-CPP-17-one-induced microsomes. Treatment of rats with 11-CH3-CPP-17-one caused a marked increase in the O-deethylation of ethoxyresorufin and, to a much lesser extent in epoxide hydrolase activity. It is concluded that (i) 11-CH3CPP 17-one is an inducer of the CYP1 family; (ii) under the present experimental conditions only the CYP1 family can oxidise the A-ring to form the 3,4-dihydroxy 11-CH3-CPP-17-one, the precursor of the ultimate carcinogen and (iii) only the CYP1 family oxidizes the diol to generate the ultimate carcinogen.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8403201 TI - Invasive tumors derived from xenotransplanted, immortalized human cells after in vivo exposure to chemical carcinogens. AB - Several chemicals that are found in cigarette smoke or diesel oil engine exhausts, such as benzo[a]pyrene (B[a]P) and 1,6-dinitropyrene (DNP) are carcinogenic in experimental animal models. In the present study, we have exposed in vivo the xenotransplanted immortalized human bronchial epithelial cell line BEAS-2B to the ultimate carcinogen of B[a]P, benzo[a]pyrene diolepoxide (BPDE), to DNP or to the benzo[e]pyrene, a less active compound that has tumor-promoting abilities in mouse skin carcinogenesis bioassays. All three compounds were administered using slow-release beeswax pellets. After a 6 month exposure, BPDE produced two tumors in seven transplants, four tumors were seen in 10 transplants treated with DNP and one tumor was observed in five tracheal grafts exposed to B[a]P. All the neoplasms were well-differentiated invasive adenocarcinomas. Tracheal transplants exposed to beeswax without carcinogen did not show any evidence of neoplastic growth, and their luminal surfaces were lined by a single or double layer of cuboidal cells. All lines derived from the adenocarcinomas showed increased in vitro resistance to serum-induced terminal differentiation, gelatinolytic activity, s.c. tumorigenicity and invasive growth in an in vivo assay. When these cell lines were compared with previously described tumor cell lines derived from xenotransplants exposed to cigarette smoke condensate, it became clear that the latter exhibited a more aggressive invasive behavior. Nevertheless treatment with the three chemicals gave rise to tumor cell lines that exhibited a similar invasive behavior in vivo, and were able to penetrate early into the wall of the tracheal transplants in which they were seeded. These data indicate that this system based on xenotransplanted bronchial epithelial cells is a very relevant model to identify human carcinogens and to study mechanisms of bronchogenic cancer pathogenesis. PMID- 8403202 TI - Differential binding of chromium(VI) and chromium(III) complexes to salmon sperm nuclei and nuclear DNA and isolated calf thymus DNA. AB - The binding of CrCl3.6H2O, Cr(NO3)3.9H2O, [Cr(L-His)2] (NO3)3.H2O, [Cr(L-Cys)(L His)].3.5H2O, [Cr(L-His)(D-Pen)].H2O, Na[Cr(L-Cys)2].2H2O, K2[Cr(GS)2].3H2O, Na2 CrO4.4H2O, and Na2Cr2O7.2H2O to salmon sperm nuclei and nuclear DNA was determined. The Cr(III)-amino acid complexes and Cr(VI) exhibited significantly lower Cr-nuclei and Cr-DNA binding levels relative to the inorganic complexes CrCl3.6H2O and Cr(NO3)3.9H2O. The binding of CrCl3.6H2O, Cr(NO3)3.9H2O and Na2Cr2O7.2H2O to salmon sperm nuclei and nuclear DNA in the presence of rat lung cytosol was determined under the same conditions. For those complexes studied in both buffer and cytosol, the Cr-DNA binding levels for Cr(III) complexes were higher in buffer than in cytosol, while a relatively higher binding level was observed for Cr(VI) in cytosol than in buffer. Slightly lower nuclear protein levels were present in Cr(VI) incubations than in Cr(III) incubations with nuclei both in the presence and the absence of cytosol. The relative binding of CrCl3.6H2O, Cr(NO3)3.9H2O, [Cr(L-His)2](NO)3.H2O, [Cr(L-Cys) (L-His)].3.5H2O, [Cr(L-His)(D-Pen)].H2O, Na[Cr(L-Cys)2].2H2O and Na2CrO4.4H2O to isolated calf thymus DNA in buffer was also determined. Positively charged, labile inorganic Cr(III) complexes, CrCl3.6H2O and Cr(NO3)3.9H2O, exhibited higher binding to DNA than [Cr(L-His) (D-Pen)].H2O, and no binding to DNA was observed with Cr(VI) and the other neutral, positively and negatively charged, inert Cr(III)-amino acid complexes. Although labile aquo chromium(III) complexes are quite reactive with DNA, the reactivity of chromium(III), formed upon intracellular reduction of carcinogenic chromium(VI), toward DNA will be diminished by complexation with cellular proteins, peptides and amino acids. PMID- 8403203 TI - Persistence, gestation stage-dependent formation and interrelationship of benzo[a]pyrene-induced DNA adducts in mothers, placentae and fetuses of Erythrocebus patas monkeys. AB - Since DNA adducts have been detected in the placentae of pregnant women who smoke cigarettes, the importance of these adducts as biomarkers of fetal exposure and risk has been evaluated using a non-human primate as a model. Pregnant Erythrocebus patas monkeys on days 50, 100 or 150 of gestation (term = 160 +/- 5 days) were treated once with 5-50 mg/kg benzo[a]pyrene (B[a]P), p.o. Fetuses were removed by Cesarean section 1-50 days after treatment and analyzed for DNA adducts by the nuclease P1 version of the 32P-postlabeling method. B[a]P induced high levels of DNA adducts in all fetal organs, placentae and maternal livers in all three trimesters of gestation. DNA adduct levels were higher in mid-gestation compared to early and late gestation. The major adduct detected was 10 beta (deoxyguanosin)-N2-yl-7 beta,8 alpha,9 alpha-trihydroxy-7,8,9,10- tetrahydro B[a]P. The adduct levels in fetal tissues increased with B[a]P dose, but at a much lower rate than in placentae or maternal livers. Preference in binding to DNA of various fetal organs was more apparent in early gestation compared to late gestation and at lower doses compared to higher doses. During early gestation and at low doses, B[a]P produced a similar level of DNA adducts in fetal lung, fetal liver, maternal liver and placenta. Individual fetal organ adduct levels correlated significantly with placental adduct levels, indicating placental and/or maternal contribution to genotoxic injuries in fetuses. However, the slopes of linear regression lines of correlation analyses varied among organs and among gestation stages at treatment, indicating fetal contribution to its own genotoxic injuries. DNA adduct levels in fetal skin were the lowest of all fetal organs tested and less affected by gestational stages at time of treatment. In contrast, DNA adduct levels in fetal liver exhibited distinct gestation stage specificity with higher adduct levels attained during mid-gestation compared to other stages of gestation. Adduct levels decreased at a much faster rate during the first 10-15 days compared to 15-50 days after B[a]P treatment. However, 10% of DNA adducts persisted 50 days after treatment in all organs studied. Together, the results suggest that placental adduction accurately indicates fetal exposure. Toxicokinetics of B[a]P and its metabolites as well as maternal, placental and fetal competence in activation and deactivation of B[a]P may be critical determinants in overall fetal risk to genetic damage. Importantly, maximal sensitivity to transplacental DNA damage may be during mid-gestation. PMID- 8403204 TI - Relationship between the GSTM1 genetic polymorphism and susceptibility to bladder, breast and colon cancer. AB - Mammalian cytosolic glutathione S-transferases (GSTs; EC 2.5.1.18) form a supergene family consisting of four distinct families, named alpha, mu, pi and theta. In humans one member of the mu class gene family (GSTM1) has been shown to be polymorphic and is only expressed in 55-60% of individuals. Previous studies have shown a possible link with the null phenotype and susceptibility to cancer, in particular to lung cancer. In this study we genotyped individuals with breast, bladder and colorectal cancer. A total of 490 individuals with cancer were studied, and consisted of 97 bladder, 197 breast and 196 colorectal cancers. No significant differences were observed in the frequency of nulled individuals in bladder or breast cancer patients when compared with a control population of 225 individuals. However, a significant excess of nulled individuals were seen in colorectal cancer: 56.1% compared with the control group value of 41.8%. This was shown to be highly significant depending on the site of the tumours and > 70% of individuals with a tumour in the proximal colon were GSTM1 nulled. This is an approximately 2-fold increase in colon cancer risk in these individuals. PMID- 8403205 TI - Ploidy and nuclearity of rat hepatocytes after compensatory regeneration or mitogen-induced liver growth. AB - The distribution pattern of rat liver parenchymal cells of different ploidy classes was investigated in Wistar rats following cell proliferation induced by surgical partial hepatectomy (compensatory regeneration) or primary mitogens (direct hyperplasia). Animals were killed at 1, 2, 3, 4 and 15 days after the proliferative stimulus, and ploidy and nuclearity were measured using a computer assisted imaging system in hepatocytes isolated by collagenase perfusion. Analysis of hepatocytes from animals undergoing regeneration after partial hepatectomy revealed a large increase in tetraploid and octoploid mononucleate cells. The most striking feature was the almost complete disappearance of binucleate cells (from 20% to < 1%) at 3 days after partial hepatectomy. On the contrary, when hepatocytes were analyzed after treatment with the mitogen lead nitrate, a high number of binucleate cells (40%) was observed. The increase that was maximal at 3 days after treatment occurred mainly in 4 x 2c and in 8 x 2c compartments. This resulted in an overall increase in the ratio of binucleate/mononucleate cells as well as in the ratio (8c + 16c):(2c + 4c). The cytological changes induced by lead nitrate were not reversible 2 weeks after treatment. Because a massive elimination of excess liver cells occurred by apoptosis during this time period, it appears that polyploid cells are not preferentially eliminated. The hepatic content of DNA at the end of the regression phase was similar to control values. However, because of the higher ploidy state, the number of cells present in the liver 2 weeks after treatment appears to be lower than that of controls (approximately -16%). When liver growth was induced by a single treatment with another mitogen, the peroxisome proliferator nafenopin, a slight increase in the ploidy state of the liver was observed; because of the shift towards higher ploidy classes (8c), the increase in DNA content observed 3 days after a single treatment with nafenopin (+21%) appears to be almost entirely justified by polyploidy rather than by a hyperplastic event. PMID- 8403206 TI - Further studies on the influence of initiation dose on papilloma growth and progression during two-stage carcinogenesis in SENCAR mice. AB - The present study was designed to further evaluate the growth and progression of papillomas to squamous cell carcinomas (SCCs) in groups of animals receiving initiating doses of 7,12-dimethylbenz[a]anthracene (DMBA) producing relatively low papilloma yields following long term promotion (60 weeks) with 12-O tetradecanoylphorbol-13-acetate (TPA). For comparison, groups of animals were initiated with various doses of DMBA and then promoted with mezerein (MEZ), benzoyl peroxide (BzPo) and chrysarobin (CHRY). Following initiation, groups of female SENCAR mice received the following promoter doses: TPA (1.0 or 2.0 micrograms per mouse); MEZ (2.0 micrograms per mouse); BzPo (20.0 mg per mouse); and CHRY (52.8 micrograms per mouse). The maximum papilloma to SCC conversion ratio obtained with TPA in the current study was 0.32. This value was in the range of maximum conversion ratios obtained with the other compounds: MEZ, 0.40; CHRY, 0.32 and BzPo, 0.19. In general, the highest papilloma to SCC conversion ratios observed with TPA as the promoter were obtained in groups that received the lowest doses of DMBA and had relatively low papilloma burdens. A comparison of papilloma to SCC conversion in groups of mice promoted with TPA, MEZ or CHRY and having similar papilloma yields, revealed very similar conversion ratios. Comparison of the BzPo group with a similar papilloma yield indicated that the conversion ratio was slightly lower with this promoter. The present results indicate that in mice promoted with TPA and having relatively low papilloma numbers, a larger proportion of these papillomas progress to SCCs during continued promoter treatment. Furthermore, the results suggest that papillomas behave similarly in their ability to progress to SCCs regardless of the promoter used when comparing groups of mice with similar tumor numbers. The data are discussed in terms of possible mechanisms for the observed results. PMID- 8403207 TI - Enhancement by the tricyclic antidepressant, desipramine, of experimental carcinogenesis in rat colon induced by azoxymethane. AB - The effects of the tricyclic antidepressant drug desipramine on the incidence, number and histology of colon tumors induced by azoxymethane (AOM), and on the serum norepinephrine (NE) concentration and the labeling index of colon mucosa were investigated in Wistar rats. Rats were treated s.c. with 7.4 mg AOM/kg body wt once a week for 10 weeks, and also s.c. with 10 mg desipramine hydrochloride (desipramine)/kg body weight until the end of the experiment. Treatment with desipramine significantly increased the incidence, but not the number, of colon tumors in week 35. However, it did not influence the location and the histological appearance of the colon tumors or the histological types of colon adenocarcinomas. Furthermore, it significantly increased the serum NE level and the labeling index of colon mucosa during and after AOM treatment. These findings indicate that desipramine enhanced the development of colon tumors and that its effect may be related to its effect in increasing proliferation of colon epithelial cells. PMID- 8403208 TI - Studies on the role of topoisomerases in general, gene- and strand-specific DNA repair. AB - Using specific inhibitors we have assessed the role of topoisomerases I and II in DNA repair of the overall genome and in both strands of an essential gene, the dihydrofolate reductase (DHFR) gene in chinese hamster ovary (CHO) cells. In these studies we have: (1) used inhibitors of topoisomerases during the repair incubation and (2) studied the DNA repair in cells with altered levels of topoisomerase activity. When cells were allowed to repair after UV irradiation, the gene-specific DNA repair was not affected by either topoisomerase I or topoisomerase II inhibitors alone. However, when topoisomerase I and topoisomerase II inhibitors were added simultaneously the gene- and strand specific DNA repair were markedly inhibited. In contrast, the overall genome DNA repair was only marginally affected. This suggests that topoisomerases are involved in gene-specific DNA repair and that one type may substitute for the other in the repair process. That concept is further supported by our findings using a mutant cell line with a decreased level of topoisomerase I: gene-specific DNA repair can be inhibited by a topoisomerase II inhibitor alone. By analyzing the steady-state expression of the DHFR gene we find that inhibition of repair in the DHFR gene is not ascribed to an obvious change in the messenger level. Furthermore, using agents other than UV, we observe that the inhibitors have no effect on gene-specific repair of DNA damage introduced by the chemotherapeutic agents cisplatin and nitrogen mustard. PMID- 8403209 TI - 12-O-tetradecanoylphorbol-13-acetate-induced inhibition of gap junctional communication is differentially regulated in a transformation-sensitive Syrian hamster embryo cell line compared to early passage SHE cells. AB - The transformation-sensitive cell-line BPNi was more susceptible to 12-O tetradecanoylphorbol-13-acetate (TPA)-induced inhibition of gap junctional intercellular communication (GJIC) than early passage Syrian hamster embryo (SHE) cells, while the potency of TPA to down-regulate EGF-binding was similar in the two cell types. The kinetics of TPA-induced inhibition of GJIC suggested that different mechanisms may operate at high and low TPA concentrations. The initial inhibition after exposure to high TPA concentrations was followed by a recovery of GJIC. The recovery was much more pronounced in SHE than in BPNi cells. This effect could not be explained by differences in down-regulation of protein kinase C. Removal of high TPA concentrations also resulted in a faster recovery of GJIC in SHE than in BPNi cells. In addition, although forskolin induced a similar protection against the inhibitory effect of TPA on GJIC, forskolin restored GJIC blocked by TPA much faster in SHE than in BPNi cells. Thus, BPNi cells are more sensitive to TPA induced inhibition of GJIC than SHE cells, and have reduced capability to recover from down-regulated GJIC as compared to SHE cells. PMID- 8403210 TI - Isoenzyme shift from glucokinase to hexokinase is not an early but a late event in hepatocarcinogenesis. AB - The appearance of hepatocellular adenomas and carcinomas induced in rat liver with N-nitrosomorpholine is preceded by different types of preneoplastic foci consisting of phenotypically altered hepatocytes. The altered cells show changes in the activities of various enzymes including those of carbohydrate metabolism. Glucokinase is a type of hexokinase that is specific for hepatocytes. The enzyme plays a key role in glucose homeostasis in normal liver parenchyma and is replaced in the dedifferentiated hepatocytes of carcinomas by a low Km hexokinase. To determine the time course of the shift from glucokinase to this isoenzyme in the development of carcinomas, focal hepatic lesions were dissected from freeze-dried serial tissue sections by the laser-dissection method and studied by microbiochemical tests. In early clear and acidophilic cell foci that excessively stored glycogen (glycogenotic foci) a nearly normal glucokinase activity comparable with that of the surrounding hepatocytes was observed, whereas in the later appearing mixed cell foci a reduction in the activity of this enzyme without a compensatory increase in the hexokinase activity was found. A pronounced activity of hexokinase was only measurable in fully developed carcinomas. Since glucokinase is not modified at the post-transcriptional level, a gradual decrease in its mRNA during hepatocarcinogenesis can be assumed. A shift in gene expression from glucokinase to the isoenzyme hexokinase occurs only at the mixed cell foci/carcinoma transition step of the carcinogenic process. PMID- 8403211 TI - Molecular analysis of DNA adducts and hprt mutations produced by 6 nitrosochrysene in Chinese hamster ovary cells. AB - We have characterized the mutational spectrum of 6-nitrosochrysene in the hprt gene of Chinese hamster ovary (CHO-K1) cells and also examined the adducts formed by this compound in CHO-K1 cells by quantitative 32P-postlabeling analysis. Seventy percent of the identified mutations were simple basepair substitutions, and they occurred more often at A:T (14/17) than at G:C. Furthermore, 13 of the basepair substitutions at A:T had the mutated dA, the probable adducted residue, on the non-transcribed DNA strand. The preference for mutation at A:T contrasted sharply with the distribution of adducts formed by 6-nitrosochrysene: 80% of the identified adducts were with dG, while only 20% were probably formed through binding with dA. Analyses conducted with excision-repair-defective CHO-UV5 cells revealed both a preference for basepair substitution at A:T and an adduct profile that were similar to those found for repair-proficient CHO-K1 cells. However, basepair substitutions from CHO-UV5 cell mutants had the mutated dAs distributed randomly between the non-transcribed and transcribed DNA strands. The mutational spectra found for solvent control CHO-K1 and CHO-UV5 cells differed from those of the 6-nitrosochrysene-treated cultures. These findings suggest that 6 nitrosochrysene-induced mutations are targeted to DNA damage, but that 6 nitrosochrysene-derived dA adducts are much more effective at producing mutations than 6-nitrosochrysene-derived dG adducts. The extreme strand bias for mutated dAs in the CHO-K1 mutational spectrum appears to result from preferential removal of 6-nitrosochrysene-induced DNA lesions from the transcribed DNA strand. PMID- 8403212 TI - Induction of lung and liver tumors by fluoranthene in a preweanling CD-1 mouse bioassay. AB - Fluoranthene (FA), a major environmental pollutant, induced lung and liver tumors 6-9 months after intraperitoneal injection of 0.7, 1.75 and 3.5 mg FA into preweanling CD-1 mice. There was a dose-dependent increase in lung tumors with a maximum tumor incidence of nearly 45% and a maximum tumor multiplicity of 0.6-0.7 lung tumors/mouse. No significant difference was noted in lung tumors in the 6 and 9 month bioassays, although fewer tumors were consistently noted in mice treated with the two lowest doses of FA. Indices of lung tumor incidence (ED50) and multiplicity (TM1.0) were similar for the two bioassays and ranged from 18.9 19.5 and 26.2-27.2 mumol respectively. Male mice had up to two times more lung tumors than females but these results were not statistically significant. Liver tumors (nodular hyperplasia) appeared only in FA-treated males but no dose response relationship was evident. However, liver tumors were observed in only 0 10% of the male mice in the 6 month treatment groups, but in 20-60% of the males in the 9 month groups. Because the CD-1 preweanling mouse responded to the weak lung tumorigen FA, it is a viable, limited-term bioassay for measuring tumorigenicity of PAH and combustion emissions. PMID- 8403213 TI - In vitro DNA damage and mutations induced by a macrocyclic tetraamide chromium(V) complex: implications for the role of Cr(V) peptide complexes in chromium-induced cancers. AB - Electron paramagnetic resonance and electronic absorption spectroscopies have shown that unlike the bidentate Cr(V) complex [Cr(ehba)2O]- (ehba = 2-hydroxy-2 ethylbutanoato(2-)), I, the macrocyclic tetradentate complex, [Cr (mampa-dcb)(O)] (mampa-dcb = 5,6-(4,5-dichlorobenzo)-3,8,11,13-tetraoxo-2,2,9,9-tetrameth yl 12,12-diethyl-1, 4,7,10-tetraazacyclotridecane), II, is substitutionally inert. Low levels of DNA strand cleavage were observed after treatment with II under physiological conditions (50 mM sodium phosphate, pH 7.4, 37 degrees C) at concentrations as high as 2 mM for periods as long as 2 days. II also induces a lower number of revertants in mutation assays with Salmonella typhimurium TA100 than I when identical Cr concentrations are applied. The slopes of the linear portion of the dose-response curves are parallel, however, indicating that the mutagenicity of II is comparable to I. II is stable toward ligand exchange, reduction and disproportionation in the mutagenicity test medium and also in the presence of bacteria and the common cell reductant, glutathione. This indicates that ligand exchange with DNA and/or reduction to Cr(IV) are not responsible for the mutagenicity of II (unlike I). It is believed that II reversibly but weakly intercalates with DNA placing the Cr(V) center in close proximity for hydrogen atom abstraction or oxo-transfer reactions to ensure. This tetraamide complex is a good structural and biomimetic model for non-sulfur-containing Cr(V) peptide species that may form in vivo from reactions of Cr(VI) with peptides. Hence, it is likely to be relevant to understanding one possible mechanism by which Cr(VI) causes cancer. PMID- 8403214 TI - High copper and iron levels and expression of Mn-superoxide dismutase in mutant rats displaying hereditary hepatitis and hepatoma (LEC rats). AB - The LEC rat is a mutant strain displaying hereditary hepatitis and hepatoma. We established enzyme-linked immunosorbent assays of Cu,Zn- and Mn-superoxide dismutase (Cu,Zn- and Mn-SOD) and measured the levels of both SODs in various organs of LEC and Wistar rats. Mn-SOD concentrations were higher in LEC rats than in Wistar rats in most tissues. Cu,Zn-SOD levels of liver, kidney and intestine were lower in LEC rats than in Wistar rats. Atomic absorbtion techniques indicated that in addition to high Cu concentrations as previously reported, LEC rat livers contained high Fe concentrations relative to those in Wistar rat livers. These data suggest that increased concentrations of Fe and Cu and decreased levels of Cu,Zn-SOD may facilitate the Fenton reaction to produce hydroxyl radicals in the tissues of the LEC rat. To compensate for the decreased scavenging effects due to low levels of Cu,Zn-SOD, an adaptive increase of Mn-SOD may occur in the process of hepatitis and hepatocarcinogenesis in LEC rats. PMID- 8403215 TI - TCDD-mediated changes in hepatic epidermal growth factor receptor may be a critical event in the hepatocarcinogenic action of TCDD. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent liver tumor promoter in rats, with females being more sensitive than males. The epidermal growth factor receptor (EGFR) pathway has been implicated in altered cell growth patterns induced by tumor promoters. We investigated hepatic EGFR levels in a two-stage initiation promotion model. The TCDD doses were chosen to encompass the dose range administered in a previous chronic bioassay currently used to determine the cancer potency commonly used for human health risk assessments. TCDD was administered biweekly by oral gavage to female Sprague-Dawley rats for 30 weeks following initiation by a single dose of diethylnitrosamine (DEN). TCDD-mediated decreased EGF receptor levels were demonstrated in intact but not ovariectomized animals, consistent with previous tumor data. Likewise, previous studies have shown that TCDD induces cell proliferation in intact rats but not ovariectomized rats. We report a significant dose-dependent decrease in plasma membrane EGF receptor maximum binding capacity in both initiated and non-initiated intact rats at TCDD doses equivalent to 3.5, 10.7, 35.7 and 125 ng/kg/day. There was a significant correlation between EGF receptor effects and liver TCDD concentration. The decrease in plasma membrane EGFR determined by equilibrium binding was confirmed quantitatively by EGF stimulation of EGFR autophosphorylation as well as qualitatively by immunohistochemical detection in control and treated rats. These results demonstrate that the observed down modulation of the EGFR by TCDD is ovarian-dependent and is a sensitive effect induced at dose levels associated with TCDD hepatocarcinogenicity in rodent bioassays. PMID- 8403216 TI - Significance of selenium-labeled proteins for selenium's chemopreventive functions. AB - A 58 kDa selenium-labeled protein purified from mouse mammary epithelial cells (MMEC) was used to examine whether selenium modulates protein synthesis or is just a marker for cellular selenium status. The protein was isolated using Sephadex G150 gel filtration and DEAE-Sephadex A50 ion-exchange chromatography. It was further analysed using 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and was found as a single spot with a pI of 4.6. The immunoreactivity with anti-58 kDa antiserum and the 75Se signal co-localized on a single 58 kDa protein band on both 1D- and 2D-PAGE. Partial amino acid analysis of the peptide showed homology with the thiol protein disulfide oxidoreductase (TPDO). Varying the selenium concentration in culture medium did not affect the protein content or the immunoreactivity of the 58 kDa protein. Additionally, selenium did not seem to regulate the activity of TPDO in TM6 cells. The glutathione peroxidase activity of TM6 cells, taken as the internal positive control, was enhanced with the increase in selenium concentration in the medium. The results suggest that selenium is attached to the 58 kDa protein, but does not regulate either its protein synthesis or its functional activity. We conclude that selenium labeling of the 58 kDa protein reflects the cellular selenium status but probably is not involved in its chemopreventive ability. PMID- 8403217 TI - Quantification of specific DNA O-alkylation products in individual cells by monoclonal antibodies and digital imaging of intensified nuclear fluorescence. AB - We report the establishment of a standardized, monoclonal antibody (Mab)-based immunocytological assay (quantitative ICA) for the visualization and quantification of low levels of specific DNA O-alkylation products in individual cells by electronically intensified, indirect or direct immunofluorescence. In terms of specific binding to alkali-denatured nuclear DNA and low background noise, 10 Mabs from a collection of 154 Mabs specific for O6-methyl-2' deoxyguanosine (O6-MedGuo), O6-ethyl-2'-deoxyguanosine (O6-EtdGuo), O6-n-butyl-2' deoxyguanosine (O6-BudGuo) and O4-ethyl-2'-deoxythymidine (O4-EtdThd) with antibody affinity constants ranging between 1.0 x 10(6)-3.0 x 10(10) l/mol, were found to be best suited for ICA. At present, > or = 200 O6-EtdGuo residues (corresponding to an O6-EtdGuo/dGuo molar ratio in DNA of > or = 8.4 x 10(-8)), > or = 400 O6-BudGuo residues (O6-BudGuo/dGuo, > or = 1.7 x 10(-7)), > or = 1800 O4 EtdThd residues (O4-EtdThd/dThd, > or = 7.5 x 10(-7)) and > or = 4800 O6-MedGuo residues (O6-MedGuo/dGuo, > or = 2.0 x 10(-6)), can be quantified per diploid genome. Using a SIT video camera in combination with multiparameter image digital analysis, DNA adduct-specific rhodamine fluorescence signals are measured relative to nuclear DNA content (DAPI fluorescence). Adduct-specific fluorescence recordings in three different rat cell lines (BT3Ca, Fao and NO) were in excellent agreement with the data obtained by competitive radioimmunoassay (RIA) for hydrolysates of DNA isolated from the respective cells exposed in parallel to the same alkylating carcinogens (N-methyl-, N-ethyl- and N-[n-butyl]-N nitrosourea). Accordingly, the kinetics of O6-EtdGuo repair, as determined by ICA and RIA, respectively, were superimposable. Cell-specific, quantitative ICA can, therefore, be used for the quantification of specific, stable DNA adducts induced by alkylating carcinogens or chemotherapeutic agents and for DNA repair measurements in individual (e.g. human) cells. Work is currently underway to extend the spectrum of carcinogen--DNA adduct-specific Mabs suited for quantitative ICA. PMID- 8403218 TI - Both O4-methylthymine and O4-ethylthymine preferentially form alkyl T.G pairs that do not block in vitro replication in a defined sequence. AB - The mutagenic potential of O4-methylthymine (m4T) and O4-ethylthymine (e4T) was determined by a primer extension assay on a 25mer oligonucleotide containing a single site-specifically incorporated modified thymine. The e4T-containing oligonucleotide was prepared by using a new synthetic procedure suitable for large alkyl groups on thymine. The second-order rate constants, K(app)m and V(rel)max, permitted calculation of the frequency of formation and extension of modified base pairs compared to Watson-Crick pairing. With both m4T and e4T, the T.G type pairing was formed at least 10-fold more frequently than the nonmutagenic alkyl T.A pairing. However, there was a small but reproducible preference for m4T.G pairing. In both cases T-->C transitions would result. There was no evidence for formation of alkyl T.C or T.T. These data suggest that reported T-->A transversions by ethylation are not likely to result from O4 alkylthymine. In contrast to insertion, extension beyond alkylthymine under kinetic conditions did not occur with alkyl T.A. but only with the alkyl T.G termini. For this latter T.G type pairing, the larger ethyl group did not hinder extension compared to that of the methyl group, in the sequence studied. Under non-limiting conditions of dNTP concentration and time, complete replication could be demonstrated for both methyl- and ethyl-containing oligonucleotides. We conclude that the increase in size of the alkyl group from methyl to ethyl does not significantly affect the mutagenic potential and type of mutations of O4 alkylthymine in vitro. PMID- 8403219 TI - Production of benzoquinone adducts with hemoglobin and bone-marrow proteins following administration of [13C6]benzene to rats. AB - Adduction of hemoglobin (Hb) and bone-marrow proteins with 1,2- and 1,4 benzoquinone (1,2-BQ and 1,4-BQ) and 4,4'-diphenoquinone was examined following oral administration of [13C6]benzene to F344 rats. Linear production of [13C6]1,4 BQ adducts was observed with both Hb and bone-marrow proteins over the entire range of dosages of 0-400 mg/kg. The slopes of the regressions were 3.4 x 10(-4) (r2 = 0.997) and 1.6 x 10(-3) (r3 = 0.926) nmol/g protein/mg/kg respectively, for Hb and bone-marrow proteins. Production of [13C6]1,2-BQ adducts of Hb and bone marrow proteins also increased with benzene dosage. Although the shapes of the relationships between 1,2-BQ adducts and dosage were nonlinear, the levels were approximately 10 times greater than those associated with 1,4-BQ, suggesting a significantly greater benzene-specific dose of 1,2-BQ. Adducts of 4,4' diphenoquinone were not detected. High background levels of [12C6]adducts of 1,2 BQ and 1,4-BQ were found in Hb and bone-marrow proteins as might be expected from the many dietary sources of the phenolic precursors of the benzoquinones, i.e. phenol, catechol and hydroquinone. Background levels of the 1,2-BQ and 1,4-BQ adducts were 27.3 and 11.5 nmol/g in Hb and 44.6 and 25.6 nmol/g in the bone marrow proteins respectively. Interestingly, the production of benzene-specific adducts represented only a small fraction (< 4%) of the background levels of the same adducts. If the genotoxicity of benzene is, indeed, related to the in vivo production of BQ isomers, our results suggest that very large exposures to benzene would be needed to produce detectable increases in adduct levels and the associated risks of leukemia. PMID- 8403220 TI - Measurement of adducts of benzoquinone with hemoglobin and albumin. AB - Benzoquinones (BQ) are genotoxic species that stem from metabolism of phenolic compounds. We have developed a method for measuring adducts of 1,2- and 1,4 benzoquinone (1,2-BQ and 1,4-BQ) with cysteine residues of hemoglobin (Hb) and albumin (Alb). The method employs a reductive catalyst (Raney nickel) to selectively cleave sulfur-bound adducts so that they may be extracted with an organic solvent, derivatized with heptafluorobutyrylimidazole and measured by GC ECD or GC-MS. Reactions of 1,4-BQ (0-300 microM) with whole blood of both F344 rats and humans resulted in linear formation of adducts with Hb and Alb. Adducts of 1,2-BQ with human Alb were formed by activation of catechol (0-300 microM) in situ with horseradish peroxidase. The mean half lives in vitro of 1,4-BQ in blood from humans and rats were 3.5 and 0.68 h, respectively. Second-order rate constants for reaction of 1,4-BQ with Hb and Alb in whole blood were estimated to be 18 and 76 l/mol/h for humans and 180 and 74 l/mol/h for rats respectively. Interestingly, the following high background levels of 1,2-BQ and 1,4-BQ adducts were observed in the proteins: 1,2-BQ-human Hb = 1.5 nmol/g, rat Hb = 25.1 nmol/g; human Alb = 8.0 nmol/g; 1,4-BQ-human Hb = 27.2 nmol/g, rat Hb = 11.5 nmol/g; human Alb = 20.5 nmol/g. These large background levels of BQ adducts suggest that significant exposure to BQ precursors occurs from dietary and/or endogenous sources. Given current evidence regarding the genotoxicity of BQ, such exposures and their health consequences should be investigated. PMID- 8403221 TI - The presence of mutagens/carcinogens in the excised lung and analysis of lung cancer induction. AB - To elucidate a relationship between lung cancer and tumor induction of environmental chemicals, the presence of 1-nitropyrene (1-NP) and benzo[a]pyrene (B[a]P) in 137 Japanese (97 male and 40 female), and 21 Chinese lung specimens was examined by HPLC, and GC-MS and environmental exposure discussed. Mortality due to lung cancer in Fuyuan County, China, is much higher than that of other cancers. We investigated 21 patients who were residents of Fuyuan County. All were female aged 28-64 years and were non-smoking farmers and cooks. The histological features of the tumors were adenocarcinoma, squamous cell carcinoma and small cell carcinoma. The incidence of cancer was due to inhaling soot from the combustion of coal used for cooking and indoor heating. The polycyclic aromatic hydrocarbons, chrysene, benzo[k]fluoranthene, B[a]P, benzo[g,h,i]perylene and pyrene, but not their nitrated derivatives, were detected in substantial amounts in the resected lungs. These mutagens and carcinogens normally originate in combustion products of coal, and are discussed as the possible initiators of the tumors in the lungs of these patients. All the Japanese lung specimens were also obtained from non-smokers. The concentrations of 1-NP averaged 21.3 +/- 12.4 and 5.9 +/- 2.4 pg/g of Japanese and Chinese samples respectively. The concentrations of B[a]P averaged 180.2 +/- 103.7 and 608.7 +/- 477.1 pg/g of Japanese and Chinese samples respectively. Thus, Japanese and Chinese lung specimens were mainly contaminated by 1-NP and B[a]P, respectively. Typical tissues from a carcinomatous human lung were examined. The patient was a 64-year-old Japanese male non-smoker and farmer who had raised chickens over a period of 40 years. The histological features of the tumor were those of keratinizing squamous cell carcinoma. 1-Nitropyrene, 1-nitro-3 hydroxypyrene, 1,3-dinitropyrene and chrysene were detected at concentrations of 0.11, 0.036, 0.095 and 0.16 ng respectively per gram of lung tissue. This cancer was due to long-term exposure to the combustion products of heavy oil used in a chicken house. PMID- 8403222 TI - Low frequency of H-ras activation in naturally occurring hepatocellular tumors of C3H/HeNCr mice. AB - Previous reports from several laboratories have consistently shown that approximately 30% of spontaneous hepatocellular adenomas and 70-80% of spontaneous hepatocellular carcinomas found in aged B6C3F1 [C57BL/6 (liver tumor resistant) x C3H (liver tumor susceptible)] male mice contain one of three missense point mutations in codon 61 of the H-ras oncogene, CAA-->AAA, CGA or CTA. Irrespective of subline, the C3H mouse, the paternal parent strain of the B6C3F1 hybrid, is more susceptible to spontaneous liver tumorigenesis than the B6C3F1 mouse. However, the role of H-ras in the pathogenesis of hepatocellular tumors in C3H mice is less clear, as widely different frequencies of activation of this gene, but by the same point mutations in codon 61, have been reported by various laboratories. The present study was undertaken to characterize H-ras involvement in hepatocellular tumors of aged C3H/He mice from the NCI-Frederick Cancer Research and Development Center Colony (C3H/HeNCr). Oncogene activation was evaluated in 45 C3H/HeNCr hepatocellular tumors by the NIH 3T3 transfection assays, and point mutations in the H-ras oncogene were detected and characterized in DNA fragments amplified by PCR, using dot blot hybridization analysis with mutation-specific oligonucleotide probes and direct dideoxy sequencing of PCR products. The only transforming gene detected in these tumors by NIH 3T3 transfection was H-ras. Only 17% (1/6) of spontaneous carcinomas and 8% (3/39) of spontaneous adenomas contained transforming H-ras sequences, each with a point mutation in codon 61. In all four cases with H-ras mutations, mutated sequences comprised a minor fraction of total H-ras gene copies in DNA extracted from primary tumors. H-ras mutations thus appear to have arisen relatively late in the pathogenesis of the neoplasms. For comparison, sections of formalin-fixed, paraffin-embedded hepatocellular tumors that occurred in untreated B6C3F1 hybrid mice sired by C3H/HeNCr males were assayed for the same H-ras mutations by PCR and dot blot hybridization. Nine of 13 such tumors (4/6 carcinomas, 5/7 adenomas) were positive. The overall difference in frequency of H-ras codon 61 mutations in hepatocellular tumors in C3H/HeNCr (4/45) versus B6C3F1 (9/13) was highly significant (P = 0.000035, Fisher's exact test). These data indicate that point mutations in H-ras do not generally play a major or an initiating role in spontaneous hepatocarcinogenesis of inbred C3H/HeNCr mice and contrast with the high rate of ras mutations in liver tumors of the B6C3F1 hybrid. PMID- 8403223 TI - Comparative 32P-postlabeling analysis of benzo[a]pyrene--DNA adducts formed in vitro upon activation of benzo[a]pyrene by human, rabbit and rodent liver microsomes. AB - An interspecies comparison was made of the DNA-adducts formed in vitro upon incubation of rat liver DNA (RL-DNA) with benzo[a]pyrene (BP) in the presence of liver microsomes. Incubations were carried out with RL-DNA, BP (100 microM) and liver microsomes from hamsters, mice, rabbits, rats, 3-methylcholanthrene (3MC) pretreated rats and from humans. To analyse the adduct profiles, the 32P postlabeling technique with the nuclease P1-enhancement procedure was used. The total amount of adduct formed varied greatly with the species; also the number of adduct spots detected was different, ranging from 1 to 5. In all incubations the BP-N2-deoxyguanosine adduct was formed. Relative to the total adduct level, the level of this adduct varied from 26% with rat, 54% with hamster, 56% with 3MC pretreated rat, 58% with mouse and 75% with rabbit, to 100% with human liver microsomes. In human liver microsomes both the total amount of cytochrome P-450 per mg microsomal protein and the ethoxyresorufin O-deethylation (EROD) activity were low compared to that in animal liver microsomes. In microsomes from 3MC pretreated rats the EROD activity was strongly induced. There was no correlation between EROD activity in non-induced microsomes and total adduct level. To compare BP-DNA adduct formation in human white blood cells (WBC) with that in RL DNA, WBC were incubated with BP and 3MC-pretreated rat microsomes. The adduct profile in WBC-DNA differed from that observed after incubation of RL-DNA: the BP N2-deoxyguanosine adduct in WBC-DNA accounted for 97% of the total adduct level. It is concluded that the 32P-postlabeling method is a suitable technique to investigate both qualitative and quantitative differences in BP-DNA adduct formation between species. Furthermore, the incubation of microsomes from the liver (or other sources) with a genotoxic agent and isolated DNA or cells can be a useful approach to study the formation and stability of reactive intermediates that are able to bind to DNA, also with respect to differences between species or tissue. PMID- 8403224 TI - MNNG-induced gastric carcinoma in ferrets infected with Helicobacter mustelae. AB - N-Methyl-N-nitro-N'-nitrosoguanidine (MNNG) is a gastric carcinogen in several animal species and has been used in a number of systems to dissect the co carcinogenic potential of various compounds in the induction of gastric adenocarcinoma. Recent epidemiological evidence suggests that Helicobacter pylori may play a role as a co-carcinogen in the etiology of this tumor in humans and we have been interested in developing an animal model to study this possibility. A related organism, H. mustelae, naturally colonizes the ferret stomach and causes persistent chronic gastritis. The pathology elicited by H. mustelae in ferrets has many similarities with the human disease including different stages of multifocal atrophic gastritis which underlie the gastric ulcer and gastric carcinoma syndrome. There is little evidence, however, demonstrating the susceptibility of ferrets toward chemical carcinogenesis. We have consequently undertaken a study to ascertain whether 10 6-month-old female ferrets given a single oral dose of MNNG (50-100 mg/kg) would develop adenocarcinoma of the stomach. Five age-matched unmanipulated control animals were included for comparative purposes. All 15 ferrets were infected with H. mustelae. Nine of 10 ferrets dosed with MNNG developed gastric adenocarcinoma (29-55 months after dosing), while none of the five historical control ferrets examined an average of 63 months after the initiation of the study developed gastric tumors. By comparison, we have not observed gastric adenocarcinoma, nor has it been reported, in > 10 years of observation of untreated ferrets naturally infected with H. mustelae. The H. mustelae-infected ferret, with demonstrated susceptibility to a gastric carcinogen, plus the recent availability of specific pathogen-free ferrets, should now allow longitudinal studies in vivo to probe the role of Helicobacter in the development of gastric cancer. PMID- 8403225 TI - Differential inhibition of the DNA binding of transcription factors NF kappa B and OTF-1 by nitrogen mustard and quinacrine mustard: transcriptional implications. AB - Nitrogen mustard (HN2) and quinacrine mustard (QM) both inhibited the binding of NF kappa B to the GC-rich consensus sequence in the HIV long terminal repeat (LTR), as assessed by gel-shift assays. QM also inhibited the binding of OTF-1 to the AT-rich octamer present in the H2B promoter whereas HN2 was inactive. Inhibition of the binding of transcription factors was due to the drug interaction with DNA, since it also occurred when transcription factors were added to DNA after removal of free drug. In Jurkat cells transfected with pI3CAT, where the chloramphenicol acetyltransferase (CAT) gene is under the control of the HIV LTR, both HN2 and QM inhibited CAT gene expression. However, in Jurkat cells transfected with plasmid -147, where the CAT gene is under the control of the H2B promoter, QM inhibited CAT expression but HN2 did not. These results were obtained at concentrations of HN2 or QM that inhibited total DNA and RNA synthesis to a similar extent. The present results suggest that the more selective pharmacological activity of HN2 (HN2 is an active antineoplastic agent whereas QM is inactive and very toxic) might be related to its preferential functional inhibition of GC-rich consensus sequence, possibly important in the regulation of genes involved in the malignant proliferation and behavior of some tumors. PMID- 8403226 TI - A hypothesis for chemical carcinogen induced chromatin condensation. AB - A hypothesis for chemical carcinogen induced condensation of nuclear and nucleolar chromatin is proposed. Chromatin condensation is believed to be a result of cascade effects initiated by the inhibition of messenger RNA synthesis after exposure to chemical carcinogen in vivo. Inhibition of messenger RNA synthesis leads to the loss of protein production, which in turn causes the dephosphorylation of histone H1 and triggers the condensation of chromatin in vivo. PMID- 8403227 TI - Copper ion-mediated modification of bases in DNA in vitro by benzoyl peroxide. AB - The mouse skin tumor promoter benzoyl peroxide (BzPO), in conjunction with Cu(I), causes promutagenic damage in DNA. Because free radical intermediates are produced by the reaction of BzPO with Cu(I), we sought to determine whether BzPO plus Cu(I) caused DNA base damage typical of that caused by the hydroxyl radical. A broad range of modified DNA bases were measured by GC-MS with selected-ion monitoring after exposure of purified plasmid pCMV beta gal DNA to BzPO +/- Cu(I). Exposure to BzPO/Cu(I) caused up to 20-fold increases in the levels of adenine-derived modified bases, up to 4-fold increases in guanine- and cytosine derived modified bases, and only a < 2-fold increase in thymine-derived modified bases. The guanine-derived modified base 8-hydroxyguanine was elevated to the highest net amount, approximately 160 molecules/10(5) DNA bases. Exposure to BzPO alone or Cu(I) alone induced only minor (< < 2-fold) DNA base modification. Also, benzoic acid, the major non-radical metabolite of BzPO, or BzPO plus Fe(II) were ineffective at inducing DNA base modification. The hydroxyl radical scavenger dimethyl sulfoxide inhibited BzPO/Cu(I)-induced base modification by 10-50%. These data suggest that the reaction of BzPO with Cu(I) generates hydroxyl radical or a similarly reactive intermediate which causes DNA base damage. This damage may be responsible for BzPO/Cu(I)-mediated mutagenesis. PMID- 8403228 TI - Studies of chemopreventive effects of myo-inositol on benzo[a]pyrene-induced neoplasia of the lung and forestomach of female A/J mice. AB - There is a continuing effort at identifying chemopreventive agents that might be useful in preventing cancer of the lung. In the present study, the effects of myo inositol and dexamethasone on benzo[a]pyrene (B[a]P)-induced pulmonary adenoma formation in female A/J mice was investigated. A diet containing 3% myo-inositol fed beginning 1 week after B[a]P administration reduced the number of pulmonary adenomas by 40% but did not prevent forestomach tumors, which also occur in this experimental model. Under the same conditions, dexamethasone, 0.5 micrograms/g diet, inhibited pulmonary adenoma formation by 57% and also inhibited forestomach tumor formation to a similar extent. Feeding a diet containing both myo-inositol and dexamethasone resulted in an additive effect on the inhibition of pulmonary adenoma formation. The combination of myo-inositol plus dexamethasone produced almost identical inhibition of forestomach tumor formation to that of dexamethasone alone. The results of the present study are preliminary, but may provide a basis for future investigation into strategies for chemoprevention of pulmonary neoplasia. PMID- 8403229 TI - Hexose metabolism in pancreatic islets: succinate dehydrogenase activity in islet homogenates. AB - Succinate dehydrogenase activity was measured in rat pancreatic islet homogenates incubated in the presence of [1,4-14C]succinate, the reaction velocity being judged through the generation of 14CO2 in the auxiliary reactions catalysed by pig heart fumarase and chicken liver NADP-malate dehydrogenase. In the presence of 1.0 mM succinate, the reaction velocity averaged 5.53 +/- 0.44 pmol min-1 microgram-1 islet protein. The Km for succinate was close to 0.4 mM and the enzymic activity was restricted to mitochondria. These kinetic results indicate that, under the present experimental conditions, the activity of succinate dehydrogenase does not vastly exceed that of either NAD-isocitrate dehydrogenase or the 2-ketoglutarate dehydrogenase complex, at least when the latter enzymes are activated by ADP and/or Ca2+. Nevertheless, the activity of succinate dehydrogenase is sufficient to account for the increase in O2 uptake evoked in intact islets by the monomethyl ester of succinic acid. It could become a rate limiting step of the Krebs cycle in models of B-cell dysfunction. PMID- 8403230 TI - An in situ study of beta-glucosidase activity in normal and Gaucher fibroblasts with fluorogenic probes. AB - Beta-glucosidase activity was evaluated in situ by means of fluorogenic probes in normal human fibroblasts and fibroblasts from homozygous carriers of the Gaucher trait. Probe internalization, targeting to lysosomes and post-cleavage probe retention were the primary concerns. Internalization and targeting were attempted by in situ photosensitized labilization of lysosomal membranes, lysosomotropic detergents and the use of low density lipid (LDL) or the receptor ligand apolipoprotein E (ApoE). Post-cleavage increase of fluorescence with fluoresceinyl (bis) beta-glucopyranoside was appreciably above the rather large pre-cleavage emission. In cells incubated overnight with nonylumbelliferyl-beta glucoside (UG9) in the presence of bovine serum albumin and in the absence of ApoE, the probe was dealt with as a cytotoxic agent, accumulating in a paranuclear cap, most likely comprising elements of the endoplasmic reticulum (ER) and Golgi apparatus. Targeting of UG9 to lysosomes occurred within 1 to 3 h of preincubation in the presence of ApoE. There was some evidence of specificity, as Gaucher fibroblasts exhibited weaker cleavage of UG9 (by 50 per cent or more) compared to normal fibroblasts, but in the Gaucher cells there was some residual beta-glucosidase activity. Cleavage of UG9 was nearly totally suppressed in Gaucher cells treated with the beta-glucosidase inhibitor, conduritol B epoxide, for 24 h to 7 days. Suppression in the control fibroblasts was evident but to a lesser degree. The in situ method of fluorogenic assay established for beta glucosidase deficiency, is in principle applicable to enzyme deficiencies in other lysosomal storage diseases, or to evaluate enhanced enzyme activity following gene therapy. PMID- 8403231 TI - Stability and optimization of canalicular function in hepatocyte couplets. AB - An enriched preparation of rat hepatocyte couplets was obtained by collagenase perfusion and subsequent elutriation (> 85 per cent couplets and triplets; viability of over 95 per cent). Canalicular secretory activity (the ability to accumulate cholyl-lysyl-fluorescein, CLF) was first apparent after 2 h of culture at 37 degrees C and was present in over 80 per cent of the total population after 5-6 h. This remained almost constant for at least 4 h in both elutriated and directly plated cells. Initial storage of freshly prepared couplets at 4 degrees C for up to 6 h prior to incubation had no adverse effect upon secretory function. Reduction of canalicular secretory activity occurred at a concentration of the hepatotoxic agent menadione (IC50 17 microM) that was lower than that required to induce mild plasma-membrane blebbing (IC50 43 microM). This study has optimized and characterized the canalicular secretory effectiveness and stability of an enriched preparation of hepatocyte couplets, and established the feasibility of studies of toxic agents on hepatobiliary function in a heterogeneous population of hepatocytes. In this preparation other biochemical parameters can be assessed, thus complementing previous techniques using individual couplets. PMID- 8403232 TI - Isolation and cloning by a polymerase chain reaction of a genomic DNA fragment of the human slow skeletal troponin (TNNT1) gene. AB - The genomic 3' structure of the gene coding for the human slow skeletal troponin T (TNNT1) gene, is reported. An intron of 912 nucleotides containing an Alu element has been identified and characterized. The complexity of the sequenced region suggests an alternative exon use. The present results may be valuable for further studies on the gene structure of TNNT1 and the related troponin gene family. PMID- 8403233 TI - Effect of diets containing adenosine, guanosine, inosine or xanthosine on the nucleotide content of Artemia. Influence of mycophenolic acid. AB - Artemia uses the stored diguanosine tetraphosphate as a source of adenine and guanine nucleotides during development from the encysted gastrula to the free swimming larva. Further development of the larvae depends on a dietary source of purine rings. We have investigated the growth of Artemia in axenic cultures supplemented with 0.6 mg ml-1 of adenosine, guanosine, inosine or xanthosine. The total protein and soluble nucleotide content of Artemia grown in the presence of adenosine, guanosine or inosine was very similar, around (2 A260 units and 500 mg protein) and (4 A260 units and 1000 mg protein) after 4 and 6 days of postlarval development, respectively. The nucleotide pattern of those extracts subjected to HPLC were almost identical, the major peaks corresponding to ATP, ADP and AMP. Other nucleotides, not well characterized, were also present in those extracts. Mycophenolic acid (10 micrograms ml-1) inhibited the growth of Artemia (as measured by their protein and soluble nucleotide content) in the presence of adenosine and inosine as the purine source, and had no appreciable effect in the presence of guanosine. A quantitative analysis of the chromatographic peaks obtained from Artemia grown in the presence of any of the three nucleosides +/- mycophenolic acid showed that the effect of the antibiotic on each one of the chromatographic peaks was very similar, suggesting that Artemia, and probably other organisms as well, tend to maintain a balance between all nucleotides and to adjust the overall level to the limiting step(s) in their rates of synthesis/interconversion. Xanthosine was not able to support the development of Artemia. PMID- 8403234 TI - Differential effects of unsaturated fatty acids on phospholipid synthesis in human leukemia monocytic U937 cells. AB - The biosynthesis of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) in monocyte-like leukemia U937 cells was monitored by adding [3H]choline, [14C]ethanolamine or [14C]glycerol to the culture media; incorporation into phospholipid (PL) increased with time. The effect of unsaturated fatty acids (UFA) on PC and PE synthesis was investigated by pretreating U937 cells for 72h with 10 microM 18:1 (n - 9), 18:2 (n - 6), 18:3 (n - 3), 20:4 (n - 6) and 20:5 (n - 3). The UFA caused no alteration in cell growth, as evidenced by light microscopy and the incorporation of [3H]thymidine and [3H]leucine. Total cellular uptake of radioactive precursors remained unaffected by all the treatments. Pretreatment with 20:5 resulted in approximately 25 per cent reduction in the incorporation of [3H]choline into PL, while no significant effect was detected with the other UFAs. 18:3, 20:4 and 20:5 depressed the incorporation of [14C]ethanolamine into PL by 34 per cent, 28 per cent and 49 per cent respectively. However, there was no redistribution of label with any of the treatments. 18:3, 20:4 and 20:5 also antagonized the stimulatory effect of endotoxin (LPS) on PC and PE synthesis. In addition, the incorporation from [14C]glycerol into PC and PE was reduced by 18:3, 20:4 and 20:5. Although the PL composition of the cells remained essentially unaffected, our study shows that chronic treatment of U937 cells with n - 3 PUFA (20:5) depressed PC and PE synthesis, and 18:3 and 20:4 also caused inhibition of PE synthesis. PMID- 8403235 TI - Effect of cocaine and morphine on neutral endopeptidase activity of human peripheral blood mononuclear cells cultured with lectins. AB - We have tested the effect of alkaloids (cocaine, morphine) and enkephalins on neutral endopeptidase of peripheral blood mononuclear cells activated by lectins. When treated with concanavalin A and cocaine, peripheral blood mononuclear cells showed an enhanced activity (+110 per cent) of the membrane neutral endopeptidase, which was not related to the expression of the common acute lymphoblastic leukemia antigen at the cell surface, although both molecules have the identical amino acid sequence. Phytohemagglutinin-P, morphine and synthetic enkephalins did not induce the activity of neutral endopeptidase nor the expression of common acute lymphoblastic leukemia antigen. Our findings suggested that the drugs of abuse, cocaine and morphine, affected specific membrane constituents without altering proliferation, subcellular localization of membrane enzymes or the surface immune phenotype of peripheral blood mononuclear cells. PMID- 8403236 TI - Continuous column culture system for adherent cells. AB - A new continuous column culture system for adherent cells was developed using beads. The beads were packed in a column and an appropriate medium was continuously passed through. The whole system was kept under closed conditions. L cells and C6 cells were cultured by this new system. The number of cells increased linearly up to 16 days and reached a maximum at around 18 days. As the heat production remained constant for 16 days, it can be concluded that cells grown in this system had identical characteristics. The final concentration of cells reached was 1.0 x 10(8) ml-1. The cells could grow both in the upward and the downward direction. Advantages of this system are: (1) Cells can be recovered in their adherent form on the beads; (2) cells can easily be collected from the column by trypsinization, and (3) cells remaining in the column after trypsinization can grow again. PMID- 8403237 TI - Lectin binding and uptake and glycoprotein characterization of isolated porcine aortic endothelial and smooth muscle cells. AB - Endothelial and smooth muscle cells were isolated from porcine aorta and kept in short-term culture. To determine the terminal carbohydrate composition of the plasma membranes from both cell populations, the cells were incubated with a panel of fluorescein-labelled lectins. Both cell populations shared a number of terminal carbohydrates, but the N-galactosamine specific lectin Wistaria floribunda agglutinin labelled only endothelial cells. A lectin which selectively labelled smooth muscle cells was not found. Western blot analysis of isolated endothelial cell membrane glycoproteins indicated that most membrane glycoproteins are labelled by Wistaria floribunda agglutinin. PMID- 8403238 TI - A descriptive study of blind children in nurseries with sighted children. AB - In a longitudinal descriptive and qualitative study, nine blind pre-school children have been observed in natural interactional settings in their nurseries. The objective was to give a detailed description of the blind child's activities and social situation in a group of sighted children. The results showed that the blind child's overall behaviour in the pre-school setting was different when compared with the sighted children's behaviour, as shown in activities like orientation, exploration and play. The blind children seldom participated in sighted children's play and they seldom initiated contact with the other children. The sighted children initially showed an interest in the blind child but as their initiatives were often neglected, they were not encouraged to proceed. The teachers played an important role for the blind children, both as a visual interpreter and as a play-mate. However, many of the teachers had great difficulties in understanding the blind child's way of functioning. Alternative pre-school settings for blind children are discussed. PMID- 8403239 TI - Alcoholic parents and their children. AB - Some 211 male alcoholic in-patients were compared with a simple random sample of 200 men from Greater Stockholm. The group of male alcoholic in-patients and the random sample were subdivided with respect to alcohol consumption and use of hepatotoxic drugs: (IA) men from the random sample with low or moderate alcohol consumption and no use of drugs (n = 169); (IB) men from the random sample with low or moderate alcohol consumption with use of drugs (n = 31); (IIA) alcoholic in-patients with use of alcohol but no drugs (n = 171); (IIB) alcoholic in patients with use of alcohol and drugs (n = 40). Earlier and more severe alcohol related and anti-social problems were found among subjects with an alcoholic parent than among subjects without an alcoholic parent. The highest level of problems was noted for subjects with alcoholism in both parents and among the alcoholic in-patients. Groups which resembled each other were the drug users in the alcoholic group and in the general sample. Both inherited and environmental factors are important determinants and many of these individuals have psycho social problems as children and adults. The children of those adults who used alcohol in combination with drugs (IIB) had most problems and the most severe problems. In the general population sample, those who used alcohol in combination with drugs (IB) had so many problems in the family and psycho-social problems themselves that they resembled the alcoholic in-patients and especially the group with high alcohol consumption in combination with drugs (IIB). A new finding is that the high-risk groups IB and IIB, who used both alcohol and drugs, had experienced a more disturbed school career and were more aggressive, had more nervous problems, and were more emotionally disturbed than the other groups. It is concluded that alcohol and drug use by parents may be predictive of future alcoholism in their children. PMID- 8403240 TI - Children and adolescents with brittle bones--psycho-social aspects. AB - Osteogenesis imperfecta (OI) or brittle bones is a most unusual disease and there is a limited number of people in the whole world suffering from it. Most studies reported from this area are in the medical field and few illuminate the psycho social effects of the disease. Three studies of children and young people with brittle bones have been carried out within the psycho-social field in Sweden. They are all based on inquiries and interviews with children and parents engaged in the Swedish National Organization for Physically Disabled Children and Adolescents. PMID- 8403241 TI - Re: Disclosure of Down's syndrome: parents' recent experiences. PMID- 8403242 TI - The effect of crocetin on hemorrhagic shock in rats. AB - There is a reduction in oxygen consumption during hemorrhagic shock, and it has been suggested that this correlates with mortality. Recent data indicate that the consumption of oxygen may depend on its diffusion from the erythrocytes to the mitochondria; thus, enhancing this rate might increase tissue oxygen extraction during hypovolemia. Crocetin, a carotenoid compound which has been shown to increase oxygen diffusivity, was used in rats bled 40% of their blood volumes, and resulted in increased whole-body oxygen consumption and survival rates. Magnetic resonance spectroscopy data also indicate that crocetin increased oxygen uptake by muscle. Other factors which might account for these results, such as possible effects of crocetin on red cell deformability and mitochondrial respiration rates, were also investigated, but the mechanism of action seems to be related to the increased diffusion of oxygen through plasma. PMID- 8403243 TI - Externalization and internalization of (Na+ + K+)-ATPase in rat heart during different phases of sepsis. AB - Changes in the distribution of (Na+ + K+)-ATPase in two subcellular fractions, the sarcolemma and the light vesicle, of rat heart during sepsis were studied. Sepsis was induced by cecal ligation and puncture (CLP). The alpha-subunit of (Na+ + K+)-ATPase was photoaffinity labeled with [alpha-32P]8-N3ATP. The results show that septic rat heart exhibits hyperdynamic (hypermetabolic) phase during early (9 hr post-CLP), followed by hypodynamic (hypometabolic) phase during late (18 hr post-CLP) sepsis. Marker enzyme and beta-adrenergic receptor assays depict that the light vesicle fraction is the intracellular site of surface receptor. The incorporation of the photolabel into the alpha-subunit (M(r) = 98,000) of the (Na+ + K+)-ATPase in sarcolemmal fraction was increased by 60% (P < 0.01) during early sepsis, but was decreased by 63% (P < 0.01) during late sepsis. In contrast, the binding of 98,000-M(r) peptide in light vesicles was decreased by 40% (P < 0.01) in early sepsis, but was increased by 102% (P < 0.01) during late sepsis. The ouabain-sensitive (Na+ + K+)-ATPase activity was increased by 31% (P < 0.05) during the early sepsis, but was decreased by 32% (P < 0.01) during late sepsis in the sarcolemmal fraction; while in the light vesicle fraction, the (Na+ + K+)-ATPase activity was decreased by 21% (P < 0.01) during early sepsis, but was increased by 47% (P < 0.01) during the late phase of sepsis. The yield of membrane proteins for each specific fraction remained unchanged for control, early sepsis, and late sepsis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403244 TI - Altered levels of mRNA encoding enzymes of hepatic glucose metabolism in septic rats. AB - We investigated whether the multiple pathophysiological signals generated in a peritonitis septic model alter the mRNA levels of glycolytic and gluconeogenic enzymes, and whether these alterations are associated with glucose dyshomeostasis. Rats were sham-operated in the control group, and peritonitis sepsis was produced by a 1 cm cecal incision in the septic group. At 2, 4, and 6 hr post-surgery, total cellular RNAs were isolated from livers, and Northern blots performed to measure mRNA levels of aldolase B (ADL), lactate dehydrogenase (LDH), pyruvate kinase (PK), phosphoenolpyruvate carboxykinase (PEPCK), and glucokinase (GK). Hepatic PEPCK enzymatic activity was measured by condensing 14CO2 with phosphoenolpyruvate (PEP) to form malate. Serum glucose concentrations were also measured. We found the following: At 2 hr of peritonitis sepsis, serum glucose concentrations, mRNA levels of all enzymes, and PEPCK enzymatic activity increased over control levels. At 4 hr of peritonitis sepsis, serum glucose concentrations and mRNA levels of GK and PK continued to increase; mRNA levels of all other enzymes, as well as PEPCK enzymatic activity decreased to or below control levels. At 6 hr of peritonitis sepsis, serum glucose concentrations, mRNA levels of all enzymes, and PEPCK enzymatic activity decreased to or below control levels. We concluded that sepsis affects mRNA levels of glycolytic and gluconeogenic enzymes at the transcriptional level, and that these alterations are associated with glucose dyshomeostasis. PMID- 8403245 TI - Age-related differences in responses to endotoxin infusion in unanesthetized piglets. AB - Newborn endotoxic shock syndrome is associated with high morbidity and mortality, yet presents with different clinical manifestations than in older patients. To determine the influence of age on hemodynamic and metabolic responses to endotoxin, we developed a chronically instrumented endotoxic shock model using eight 1-3-day-old and seven 2-3-week-old piglets. Three days after surgery, 10 mg/kg of endotoxin was infused intravenously over 10 min in the younger group, and 5-10 mg/kg was given to the older animals. Two older piglets died immediately after infusion of 5 mg/kg of endotoxin, and five of the seven died within 4 hr, while all eight younger animals lived longer than 4 hr. Pulmonary artery pressure increased significantly after endotoxin in both groups, and there were no differences between groups. Systemic artery pressure and cardiac index fell by 44 +/- 10% and 70 +/- 15%, respectively, 5 min after endotoxin infusion in the older group, while these values did not change significantly in the younger group. Endotoxin infusion also caused greater elevation in pulmonary vascular resistance index in the older animals. In the later phase, which began 30 min after endotoxin, both groups displayed systemic hypotension and pulmonary hypertension, and the groups did not differ from one another in this regard. With progression of endotoxic shock, more severe metabolic acidosis developed in the older animals than in the younger animals. Plasma thromboxane B2 levels in the older group were about double those in younger piglets. Plasma 6-keto-PGF1 alpha and TNF alpha levels in both groups were similar and were significantly increased in the later phase.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403246 TI - Desensitization of rat cardiomyocyte adenylyl cyclase stimulation by plasma of noradrenaline-treated patients with septic shock. AB - The purpose of this study was to test the possibility that the mechanisms of catecholamine-induced desensitization of cardiac beta-adrenoceptor stimulation are modified in septic shock. Exposure of neonatal rat cardiomyocytes for 48 hr to plasma of noradrenaline-treated patients with septic shock led to a down regulation of beta-adrenoceptors by 35%, an increase in the level of inhibitory G protein alpha-subunits by 60%, and a decrease in isoproterenol-stimulated adenylyl cyclase activity by 50% in membranes prepared from the rat cardiomyocytes. Similar alterations were observed following pretreatment of the cells with plasma of adrenaline-treated patients with cardiogenic shock. In contrast, exposure of the cardiomyocytes to plasma of intensive care patients without shock, and to plasma of dopamine-treated patients with septic shock did not induce alterations of the cardiomyocyte adenylyl cyclase system. The dosage of the catecholamines had to be increased within the first two days of treatment in the noradrenaline-treated patients, but not in the dopamine-treated patients with septic shock. Thus, this observed tolerance to noradrenaline in the treatment of septic shock may, in part, be due to a desensitization of cardiac beta-adrenoceptor stimulation induced by the beta-adrenoceptor stimulatory effect of noradrenaline. PMID- 8403247 TI - Bacterial translocation after non-lethal hemorrhage in the rat. AB - Translocation of enteric bacteria has been suggested to compromise patients in severe catabolic stress. Mechanisms for this route of infection are not known. In this study, rats were subjected to hemorrhage without reinfusion during 60 min, total blood loss was 3.28 +/- 0.14 ml/100 g BW. Control groups consisted of sham operated animals without bleeding, and rats not operated at all. The mean number of viable bacteria found in mesenteric lymph nodes (MLN) of bled animals was 168 +/- 45 colony forming units (c.f.u./MLN), significantly higher compared to sham operated (5 +/- 3 c.f.u./MLN) and not operated (0 +/- 0 c.f.u./MLN) controls (P < 0.01). Cultures from MLN were positive in 7/9 rats after bleeding, in 3/9 of sham operated, and in 0/6 of non-instrumented control animals. No positive blood cultures were isolated. Escherichia coli was the dominant species found in MLN. A biochemical fingerprinting method (the PhP system) was used to identify translocating strains of E. coli among strains found in cecum. The method was also used to compare translocating strains between different animals. Our findings reveal that bacteria translocate to MLN after hemorrhage. Some phenotypes of E. coli strains translocate more frequently than others, suggesting that they have properties facilitating translocation. PMID- 8403248 TI - Platelet activating factor-induced microvascular permeability increases in the cat hindlimb. AB - Changes in microvascular permeability induced by platelet activating factor (PAF) were measured in the isolated, perfused cat hindlimb preparation, and compared to the effect produced by another inflammatory mediator, histamine. Permeability was assessed from changes in the protein reflection coefficient, as measured from changes in hematocrit and protein concentration resulting from microvascular fluid filtration. The findings were 1) PAF produces transient increases in permeability similar to histamine, but PAF is approximately 30 times as potent; 2) the permeability changes induced by 76 nM PAF can be totally inhibited by the specific PAF receptor blocker WEB-2086, but the blocker can only partially inhibit 380 nM PAF, a dose that produces a maximal increase in permeability; 3) Diphenhydramine (2 microM), an H1-receptor blocker, totally inhibits the transient permeability increase produced by 2 microM histamine; 4) Cimetidine (2 or 20 microM), an H2 blocker, could not inhibit this latter increase; 5) Isoproterenol (1 microM), a beta-agonist, totally inhibited the permeability increase produced by 1 microM histamine, but 10 microM isoproterenol only partially inhibited the maximal permeability increase produced by 10 microM histamine; 6) Isoproterenol could not inhibit PAF's permeability effect; and 7) PAF's effects were unchanged by depletion of white blood cells in the perfusate. These results suggest that PAF and histamine work through different pathways to increase permeability, but the final step of endothelial contraction, which opens large inter-endothelial gaps, occurs in response to both mediators. In addition, when concentrations of these inflammatory agents are sufficient to produce maximal permeability increases, as can occur in shock situations, then the permeability increases are more sustained and resistant to receptor inhibition. PMID- 8403249 TI - Transforming growth factor-beta in cardiac ontogeny and adaptation. AB - The transforming growth factor-beta (TGF-beta) superfamily comprises a set of regulatory peptides with multiple effects on cell growth and differentiation. The elaborate regulation of TGF-beta s during embryonic development of the heart, the upregulation of TGF-beta after hemodynamic stress, and the impact of TGF-beta on cardiac gene expression together imply a prominent functional role for this family of growth factors in cardiac organogenesis and hypertrophy. Basal and TGF beta-induced expression of skeletal alpha-actin, one of several genes specifically associated with developing or hypertrophied myocardium, each are contingent on transcriptional activation by serum response factor. A truncated form of the type II TGF-beta receptor, created by deletion of the cytoplasmic kinase domain, acts as a dominant suppressor of TGF-beta signal transduction in cultured cardiac muscle cells and may provide a suitable means to establish the functions of TGF-beta in vivo. PMID- 8403250 TI - Mouse model of arterial injury. AB - In the present study, we established an injury model of the mouse carotid artery. Complete removal of the endothelium was achieved with a flexible wire. A platelet monolayer covered the denuded surface, and damage to underlying medial smooth muscle cells (SMCs) was detected. Injection of [3H]thymidine was used to determine the replication index for medial SMCs, which was found to be 1.6% at 2 days after denudation and 9.8% at 5 days. SMCs were observed in the intima by day 8 (replication index, 66%), and by 2 weeks the intimal lesion had a similar cell content as the media. In most animals, repair of the endothelial lining was complete 3 weeks after injury. The present model will allow us to use transgenic animals to address questions relevant to vascular biology and atherosclerosis. PMID- 8403251 TI - In vivo adenoviral vector-mediated gene transfer into balloon-injured rat carotid arteries. AB - We studied the ability of adenoviral vectors to achieve gene transfer into injured arteries. A recombinant adenoviral vector expressing a nuclear-targeted beta-galactosidase gene was constructed and infused into balloon-injured rat carotid arteries. Three days after gene transfer, recombinant gene expression was assessed quantitatively by (1) measuring beta-galactosidase antigen and activity in tissue extracts and (2) histochemical staining and counting of cells expressing beta-galactosidase. Exposure of injured carotid arteries to increasing concentrations of the vector (10(8) to 10(10) plaque-forming units per milliliter) resulted in a dose-responsive increase in beta-galactosidase expression, with peak expression of approximately 43 mU or 25 ng beta galactosidase per vessel. Microscopic examination of histochemically stained arteries demonstrated gene transfer limited to the vascular media; transduced cells were identified immunohistochemically as smooth muscle cells. Counting of both histochemically stained and total nuclei in the media revealed that approximately 30% of the cells in the media of the injured vessels were transduced. Calculations based on both counting cells and on the level of beta galactosidase expression in tissue extracts suggested the presence of 5000 to 10,000 transduced cells per 10 mm of vessel. Arteries infused with either vehicle only, a control adenoviral vector, or liposomes combined with the vector plasmid contained little or no evidence of beta-galactosidase expression. High levels of in vivo beta-galactosidase expression persisted for at least 7 days after gene transfer but declined significantly by day 14. We conclude that adenoviral vector mediated gene transfer into the injured rat carotid artery results in efficient gene transfer into the vascular media, with levels of recombinant protein production significantly higher than any previously reported in arterial gene transfer studies. Adenoviral vectors appear to be particularly useful agents for in vivo arterial gene transfer. PMID- 8403252 TI - Effects of thapsigargin and cyclopiazonic acid on twitch force and sarcoplasmic reticulum Ca2+ content of rabbit ventricular muscle. AB - Thapsigargin (TG) and cyclopiazonic acid (CPA) are reported to be specific high affinity inhibitors of the sarcoplasmic reticulum (SR) Ca2+ pump in isolated membranes and cells, with TG causing complete pump inhibition at nanomolar concentrations. To evaluate the effectiveness of TG and CPA in small multicellular cardiac preparations, we used rapid cooling contractures (RCCs) to assess the SR Ca2+ load. In contrast to observations in single myocytes, TG caused remarkably slow and incomplete SR Ca2+ depletion in multicellular preparations. A 45-minute exposure to 500 microM TG at 30 degrees C and 0.5-Hz stimulation only decreased RCCs by 76 +/- 5% (and 100 microM CPA reduced RCCs by 59 +/- 10% [mean +/- SEM]). In contrast, 10 minutes with 20 mM caffeine completely abolished RCCs. This confirms that there was still a caffeine sensitive pool of Ca2+ in the TG-treated muscle. The time constant of rest decay of RCCs was accelerated by both TG (from 83 +/- 18 to 26 +/- 6 seconds) and CPA (from 68 +/- 11 to 10 +/- 5 seconds). This might be expected since Ca2+ leaking from the SR during rest cannot be taken back up as efficiently, favoring Ca2+ extrusion by the sarcolemmal Na(+)-Ca2+ exchanger. TG and CPA decreased twitch force (by 44 +/- 7% and 40 +/- 11%, respectively) and increased twitch duration, presumably because of the SR effects. We conclude that complete blockade of SR Ca2+ uptake by TG or CPA in multicellular preparations cannot be assumed, even at high [TG] or [CPA], unless evaluated (eg, by RCC). PMID- 8403253 TI - Sarcoplasmic reticulum Ca2+ uptake and thapsigargin sensitivity in permeabilized rabbit and rat ventricular myocytes. AB - Ca2+ uptake by the sarcoplasmic reticulum (SR) and free [Ca2+] were measured simultaneously with indo 1 and a Ca(2+)-selective minielectrode in suspensions of permeabilized rabbit or rat ventricular myocytes (approximately 10 mg/mL protein). In the presence of 25 mumol/L ruthenium red and 10 mmol/L oxalate, the Km for Ca2+ uptake by the SR was approximately 250 nmol/L in rabbit and rat ventricular myocytes. The maximal Ca2+ uptake rate was 2.4 times higher in rat than in rabbit. Addition of 5 nmol thapsigargin (TG) per milligram cell protein abolished Ca2+ uptake completely in both species. The [TG] necessary for a half maximal reduction of the uptake rate (K1/2) was 55 pmol/mg cell protein for rabbit and 390 pmol/mg cell protein for rat. Assuming that the number of pump sites is two times the concentration of TG necessary to inhibit half of the Ca2+ pump activity (ie, the TG affinity is very high), the density of pump sites is 7.7 mumol/kg wet wt for rabbit and 54.6 mumol/kg wet wt for rat. Despite a fivefold decrease of the Ca2+ uptake rate by a submaximal [TG], the permeabilized myocytes were still able to lower the free [Ca2+] to < 150 nmol/L from a peak value > 10 mumol/L. The relative inhibition of Ca2+ uptake by TG did not depend on the free [Ca2+]. Addition of more than 5 nmol TG per milligram cell protein abolished Ca2+ uptake by the SR completely in < 15 seconds and reduced the uptake rate by 95% in 5 seconds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403254 TI - Chronic exercise enhances endothelium-mediated dilation of epicardial coronary artery in conscious dogs. AB - Whether endothelium-derived relaxing factor (EDRF)/nitric oxide (NO) plays a role in the dilation of the left circumflex coronary artery during acute exercise and whether endothelium-mediated dilation of this artery is altered after chronic exercise training have not been determined previously. Nine dogs were chronically instrumented for measurements of systemic hemodynamics, left circumflex coronary artery diameter, and blood flow. Acute treadmill exercise (10.9 km/h) caused dilation of the circumflex coronary artery by 4.33 +/- 0.84% and an increase in coronary blood flow by 32 +/- 5.2 mL/min. After the administration of intravenous nitro-L-arginine, the dilation of the circumflex coronary artery was converted to vasoconstriction (-4.13 +/- 1.58%), whereas the increase in coronary blood flow was not altered (24 +/- 3.6 mL/min). Chronic exercise training (2 hours each day at a speed of 10.9 km/h for 7 days) enhanced acetylcholine-induced dilation and reactive dilation (following release of a brief coronary artery occlusion) of the large coronary artery (P < .05), whereas the coronary blood flow responses were not changed. These enhanced acetylcholine-induced and reactive dilations of the circumflex coronary artery were due to a greater release of EDRF/NO since they were eliminated by nitro-L-arginine. Thus, in the circumflex coronary artery, EDRF/NO-dependent dilation was enhanced after 7 days of exercise training. This may represent the mechanism responsible for the perception that chronic exercise induces cardiovascular "well being." PMID- 8403255 TI - Rapid turnover of the AMP-adenosine metabolic cycle in the guinea pig heart. AB - The intracellular flux rate through adenosine kinase (adenosine-->AMP) in the well-oxygenated heart was investigated, and the relation of the AMP-adenosine metabolic cycle (AMP<-->adenosine) to transmethylation (S-adenosylhomocysteine [SAH]-->adenosine) and coronary flow was determined. Adenosine kinase was blocked in isolated guinea pig hearts by infusion of iodotubercidin in the presence of the adenosine deaminase blocker erythro-9-(2-hydroxy-3-nonyl)adenine (5 mumol/L). Iodotubercidin (1 nmol/L to 4 mumol/L) caused graded increases in venous effluent concentrations of adenosine, from 8 +/- 3 to 145 +/- 32 nmol/L (mean +/- SEM, n = 3), and in coronary flow, which increased to maximal levels. Flow increases were completely abolished by adenosine deaminase (5 to 10 U/mL). Interstitial adenosine concentrations, estimated using a mathematical model, increased from 22 nmol/L during control conditions to 420 nmol/L during maximal vasodilation. The possibility that iodotubercidin caused increased venous adenosine by interfering with myocardial energy metabolism was ruled out in separate 31P nuclear magnetic resonance experiments. To estimate total normoxic myocardial production of adenosine (AMP-->adenosine<--SAH), the time course of coronary venous adenosine release was measured during maximal inhibition of adenosine kinase with 30 mumol/L iodotubercidin. Adenosine release increased more than 15-fold over baseline, reaching a new steady-state value of 3.4 +/- 0.3 nmol.min-1 x g-1 (n = 5) after 4 minutes. In parallel experiments, the relative roles of AMP hydrolysis and transmethylation (SAH hydrolysis) were determined, using adenosine dialdehyde (10 mumol/L) to block SAH hydrolase. In these experiments, adenosine release increased to similar levels of 3.4 +/- 0.5 nmol.min-1 x g-1 (n = 6) during inhibition of adenosine deaminase and adenosine kinase. It is concluded that (1) maximal increases in coronary flow are elicited by increases in interstitial adenosine concentration to approximately 400 nmol/L, (2) more than 90% of the adenosine produced in the heart is normally rephosphorylated to AMP without escaping into the venous effluent, (3) AMP hydrolysis is the dominant pathway for cardiac adenosine production under normoxic conditions, and (4) the high rate of adenosine salvage is due to rapid turnover of a metabolic cycle between AMP and adenosine. Rapid cycling may serve to amplify the relative importance of AMP hydrolysis over transmethylation in controlling cytosolic adenosine concentrations. PMID- 8403256 TI - Use-dependent block and use-dependent unblock of the delayed rectifier K+ current by almokalant in rabbit ventricular myocytes. AB - A voltage-clamp analysis of the effect of almokalant on the delayed rectifier K+ current (IK) was made in rabbit ventricular myocytes. The two-suction pipette method was used, and appropriate voltage-clamp protocols were used to study more specifically use dependence, block development, and recovery from block. Almokalant interacted with the IK in two ways: it shifted the activation curve in the hyperpolarizing direction (stimulatory effect) and blocked the open IK channel in a use-dependent way (inhibitory effect). For 2-second voltage clamps to +20 mV, half-maximum block was obtained at 5 x 10(-8) mol/L, with a Hill coefficient of 1.76. Use-dependent block was related to an open-channel block that occurred at 0 mV with a time constant of 1.07 second and a rather slow recovery from block: at -50 mV, recovery time constant was approximately 10 seconds; at -75 mV, recovery was practically absent. The absence of an important recovery at negative membrane potentials is consistent with the hypothesis of the drug being trapped in the channel. A limited frequency-dependent block could be demonstrated. Use-dependent unblock was demonstrated by a rapid recovery from block during stimulation following complete washout of the drug. It is concluded that almokalant shifts the activation curve of IK in the hyperpolarizing direction, blocks the open channel, and is trapped by the closure of the activation gate. PMID- 8403257 TI - Impact of alpha-adrenergic coronary vasoconstriction on the transmural myocardial blood flow distribution during humoral and neuronal adrenergic activation. AB - Increased heart rate and left ventricular pressure during humoral and neuronal adrenergic activation act to restrict blood flow preferentially in the subendocardium. The hypothesis was advanced that alpha-adrenergic coronary vasoconstriction preferentially in the subepicardium may counterbalance the enhanced extravascular compression in the subendocardium and serve to maintain blood flow transmurally uniform. In 40 anesthetized dogs, regional myocardial blood flow was determined with colored microspheres; wall function, with sonomicrometry. Humoral adrenergic activation (HAA) was induced by a combination of intravenous atropine, intravenous norepinephrine, and atrial pacing during baseline coronary vasomotor tone (group 1, n = 6) and in the presence of maximal coronary vasodilation with intravenous dipyridamole (group 2, n = 6). In an additional group, HAA was induced by intravenous norepinephrine in the presence of dipyridamole but without atropine and atrial pacing in order to increase end diastolic left ventricular pressure (group 3, n = 6). Measurements were performed at rest, during HAA, and during ongoing HAA with the intracoronary infusion of the alpha-antagonist phentolamine (Phen). At unchanged mean aortic pressure, Phen improved blood flow particularly to the inner layers as follows: from 1.42 +/- 0.40 (mean +/- SD) to 1.90 +/- 0.40 mL/(min.g) (group 1, P < .05), from 4.99 +/- 2.31 to 5.53 +/- 2.56 mL/(min.g) (group 2, P < .05), and from 6.01 +/- 1.41 to 6.29 +/- 1.27 mL/(min.g) (group 3, P < .05), associated with a decrease in outer layer blood flow in groups 2 and 3. In 16 additional dogs, beta-adrenoceptors were blocked by propranolol and muscarinic receptors by atropine. Neuronal adrenergic activation (NAA) was induced by cardiac sympathetic nerve stimulation (CSNS) during baseline coronary vasomotor tone (group 4, n = 8) and in the presence of maximal vasodilation (group 5, n = 8). Measurements were performed at rest, during a first CSNS, and 20 minutes later during a second CSNS+Phen. The reproducibility of two consecutive episodes of CSNS 20 minutes apart was demonstrated in a separate set of experiments (n = 6). At matched mean aortic pressures, Phen improved blood flow to all myocardial layers in group 4, whereas in group 5, Phen induced a redistribution of myocardial blood flow toward subepicardial layers [from 4.44 +/- 0.96 to 4.81 +/- 0.83 mL/(min.g), P < .05] at the expense of inner layers. With the addition of Phen, there was no change in regional wall function in any group of dogs studied. Thus, during HAA, alpha adrenergic coronary vasoconstriction does not exert a beneficial effect on transmural blood flow distribution. During NAA, a beneficial effect of alpha adrenergic coronary vasoconstriction becomes apparent only under conditions of maximal coronary vasodilation. PMID- 8403258 TI - Contribution of endogenous endothelin-1 to the progression of cardiopulmonary alterations in rats with monocrotaline-induced pulmonary hypertension. AB - Endothelin-1 (ET-1) is known to have potent contractile and proliferative effects on vascular smooth muscle cells and is known to induce myocardial cell hypertrophy. We studied the pathophysiological role of endogenous ET-1 in rats with monocrotaline-induced pulmonary hypertension. Four-week-old rats were given a single subcutaneous injection of 60 mg/kg monocrotaline (MCT rats) or saline (control rats) and were killed after 6, 10, 14, 18, and 25 days. In the MCT rats, right ventricular systolic pressure progressively increased and right ventricular hypertrophy developed in a parallel fashion. The venous plasma ET-1 concentration also progressively increased, and this increase preceded the development of pulmonary hypertension. The isolated pulmonary artery exhibited a significantly weaker response to ET-1 in the MCT rats on day 25 but not on days 6 and 14. In the MCT rats, the expression of prepro ET-1 mRNA as measured by Northern blot analysis significantly increased in the heart on days 18 and 25, whereas it gradually decreased in the lungs. The peptide level of ET-1 in the lungs also significantly decreased in the pulmonary hypertensive stage. The expression of prepro ET-1 mRNA had increased by day 6 only in the kidneys. Continuous infusion of BQ-123, a selective ETA receptor antagonist, by an osmotic minipump (14.3 mg per day per rat for 18 days) significantly inhibited the progression of both pulmonary hypertension (right ventricular systolic pressure, 77.8 +/- 4.2 [mean +/- SEM] mm Hg [n = 10] versus 52.3 +/- 2.4 mm Hg [n = 7]; P < .01) and right ventricular hypertrophy (right ventricle/[left ventricle +/- septum], 0.56 +/- 0.03 [n = 10] versus 0.41 +/- 0.02 [n = 7]; P < .01). Histological examination revealed that BQ-123 also effectively prevented pulmonary arterial medial thickening. The inhibition of right ventricular hypertrophy by BQ-123 may be partly ascribed to the blockade of excessive stimulation of the heart by ET-1, in addition to the prevention of pulmonary hypertension. The present findings suggest that endogenous ET-1 contributes to the progression of cardiopulmonary alterations in rats with MCT-induced pulmonary hypertension. PMID- 8403259 TI - Hypertensive rats produced by in vivo introduction of the human renin gene. AB - We established an efficient and nontoxic in vivo gene transfer method mediated by the Sendai virus (hemagglutinating virus of Japan [HVJ]), liposomes, and nuclear protein. In this study, to produce a hypertensive model rat that is dependent on human renin, the human renin gene was introduced into adult rat liver by our efficient in vivo gene transfer method using HVJ and liposomes (HVJ-liposomes). The rats treated with HVJ-liposomes containing the human renin gene showed a significant elevation of blood pressure for 6 days compared with control rats, which received injections of HVJ-liposomes without the human renin gene. On day 5 after the transfer, human active renin as well as angiotensin II were found in the plasma of rats in which the human renin gene was introduced. Moreover, the blood pressure of these rats was significantly correlated with the plasma levels of human active renin and angiotensin II. To confirm that the elevated blood pressure was due to the expression of the human renin gene, we administered a newly developed specific human renin inhibitor, FK 906. The elevated blood pressure was normalized by the intravenous administration of this drug. These data indicate that this hypertensive rat was produced by the in vivo transfer of the human renin gene into rat liver and that the expressed human renin cleaved rat substrate (angiotensinogen). This hypertensive rat produced by in vivo gene transfer should be useful in further studies on hypertension. PMID- 8403260 TI - Renal afferents signaling diuretic activity in the cat. AB - Mechanoreceptors and chemoreceptors have been identified inside the kidney, but their functional role is still largely unclear. The aim of this study was to investigate whether changes in urine output could modify the discharge rate of renal afferent fibers. Experiments were performed in anesthetized cats in which afferent renal nerve activity (ARNA) was recorded by standard electrophysiological techniques from a centrally cut renal nerve. Arterial pressure, renal blood flow velocity, urine flow rate, and renal pelvic pressure were also measured. Three diuretic maneuvers were tested in the same cat: intravenous administration of physiological saline (8 to 13 mL/min for 2 minutes), furosemide (1 mg/kg), and atrial natriuretic peptide (ANP, 1 microgram/kg). The three maneuvers increased urine flow rate and pelvic pressure, respectively, 137.0 +/- 20.6% and 136.8 +/- 21.1% (saline), 148.6 +/- 31.7% and 139.6 +/- 43.5% (furosemide), and 75.9 +/- 7.9% and 62.1 +/- 21.2% (ANP) at the time of the maximum response. Arterial pressure slightly increased after saline, did not change after furosemide, and slightly decreased after ANP. Renal blood flow increased after saline and did not change after furosemide and ANP. The three maneuvers increased ARNA by 98.4 +/- 15.2% (saline), 270.7 +/- 100.8% (furosemide), and 59.6 +/- 23.4% (ANP). Changes in ARNA significantly correlate with changes in both pelvic pressure and urine flow rate. Our data demonstrate that increments in urine flow rate increase the firing rate of renal afferent fibers and suggest that (1) pelvic pressure is the major determinant of the neural response, and (2) this increased afferent discharge is due to activation of renal mechanoreceptors. PMID- 8403261 TI - Microscopic conduction in cultured strands of neonatal rat heart cells measured with voltage-sensitive dyes. AB - Microscopic discontinuities in electrical activation were assessed in synthetic strands of neonatal rat myocytes cultured on a growth-directing matrix. An optical method using voltage-sensitive dye (RH-237) and a photodiode technique was used for recordings of membrane potential changes with subcellular resolution. Spatial resolution of the method (diameter of measurement area, 5.5 microns; interdiode distance, 30 microns) allowed for simultaneous measurements of cytoplasmic conduction time within a single cell and junctional conduction time across the cell border. In one-dimensional cell chains, where cells were juxtaposed by end-to-end connections but devoid of lateral connections, propagation of the excitation wave was strongly nonuniform: cytoplasmic conduction time was 38 +/- 30 (mean +/- SD) microseconds (n = 37), whereas junctional conduction time was 118 +/- 40 microseconds (n = 27, P < .0001). A mean delay introduced by a single junction was 80 microseconds, or 51% of conduction time. In two-dimensional strands consisting of several cells in width, which exhibited lateral as well as end-to-end connections, inhomogeneity of conduction was smaller: the cytoplasmic and junctional conduction times were 57 +/- 30 (n = 46) and 89 +/- 40 (n = 48) microseconds, respectively (P < .0001); mean junctional conduction delay was 32 microseconds (22% of conduction time). Mathematical modeling suggested that the averaging effect of lateral connections is caused by lateral convergence of local excitatory current beyond and lateral divergence before end-to-end connections. Our results demonstrate that the current flow through lateral cell-to-cell connections smooth the excitation wave front during longitudinal conduction in myocardial tissue. PMID- 8403262 TI - Intracellular myoglobin loading worsens H2O2-induced, but not hypoxia/reoxygenation-induced, in vitro proximal tubular injury. AB - Intracellular iron reportedly mediates many forms of tissue injury, including ischemic and myohemoglobinuric acute renal failure. This action may be explained by the ability of iron to catalyze the formation of the highly toxic hydroxyl radical (.OH) from H2O2 via the Fenton/Haber-Weiss reactions. To assess whether renal tubular myoglobin/iron loading, induced by a physiological mechanism (endocytosis), alters its susceptibility to O2 deprivation/reoxygenation- and H2O2-mediated injury, rats were infused with myoglobin or its vehicle (5% dextrose, control rats), and after 2 hours, proximal tubular segments (PTSs) were isolated for study. This infusion caused substantial myoglobin endocytic uptake (approximately 25 micrograms/mg PTS protein), and it doubled PTS catalytic iron content (assessed by bleomycin assay). Nevertheless, PTS viability (percent lactate dehydrogenase release) was minimally affected (4% to 6% increase), and an increased .OH burden (assessed by the salicylate trap method) did not appear to result. Deferoxamine addition, reported to protect against in vivo acute renal failure, paradoxically increased .OH levels (approximately 25%) in myoglobin loaded, but not control, PTSs. Conversely, dimethylthiourea (an .OH scavenger) depressed .OH (by approximately 80%) in all PTSs. Myoglobin/iron loading modestly increased PTS vulnerability to exogenous H2O2 addition (P < .001). However, tubular susceptibility to hypoxia (15 and 30 minutes)/reoxygenation injury was not affected. .OH levels appeared to fall in response to both forms of injury, suggesting decreased .OH production and/or .OH scavenging. To assess whether myoglobin decreases .OH levels in the presence of Fenton reactants, myoglobin and six other test proteins were incubated with Fe2+/H2O2. Myoglobin decreased .OH levels by approximately 70%, a significantly greater decrement than was observed with the other proteins tested.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403263 TI - Changes in creatine kinase M localization in acute ischemic myocardial cells. Immunoelectron microscopic studies. AB - To clarify the changes in creatine kinase M localization along with the progress of myocardial ischemia, immunoelectron microscopic studies were performed using rabbit anti-canine creatine kinase M Fab'-horseradish peroxidase conjugate in 21 dogs. Myocardial ischemia was induced by occlusion of the left anterior descending coronary artery for 15 (n = 5), 30 (n = 5), 60 (n = 5), or 180 (n = 4) minutes. Two dogs were used as normal controls. As we have already demonstrated, most creatine kinase M in normal myocardial cells was localized over the entire A band in association with the thick filament, suggesting that creatine kinase in this region (A-band creatine kinase) was the enzyme coupled with myosin ATPase. After 15 minutes of ischemia, creatine kinase M showed only minimal changes in its location, indicating that A-band creatine kinase still has the ability to couple with myosin ATPase (reversible injury). However, after 30 minutes of ischemia, A-band creatine kinase diffused markedly to the I band (transitional phase), and after 60 minutes of ischemia, it leaked out to extracellular spaces (irreversible injury). After 180 minutes of ischemia, most A-band creatine kinase disappeared from the myocardial cells (coagulation necrosis). These features of creatine kinase M localization seemed to reflect each stage of ischemic cell injury. We conclude that myocardial ischemia results in a dissociation of creatine kinase molecules from the thick filament, which leads the energy transport system to destruction. PMID- 8403264 TI - Differential downregulation of beta 2-adrenergic receptors in tissue compartments of rat heart is not altered by sympathetic denervation. AB - With agonist stimulation, cardiac beta 2-adrenergic receptors (beta 2ARs) are downregulated to a much greater extent than are beta 1ARs. It has been hypothesized that this effect is due to sympathetic innervation inhibiting the downregulation of beta 1ARs. To test this hypothesis, the technique of coverslip autoradiography was used to localize and quantify beta 1AR and beta 2AR subtypes in tissue compartments of the heart in rats subjected to sympathetic denervation by two intravenous injections of 6-hydroxydopamine (50 mg/kg per dose). After denervation, the rats were infused with L-isoproterenol (400 micrograms.kg-1 x h 1 for 7 days) or vehicle (0.001N HCI) by implantation of osmotic minipumps. Sections were incubated with 70 pmol/L of the beta AR antagonist [125I]iodocyanopindolol (ICYP) alone or in the presence of 5 mumol/L DL propranolol or 5 x 10(-7) mol/L CGP 20712A (a beta 1AR antagonist). Binding of ICYP to sections of rat hearts was saturable and stereoselective and was displaced by beta AR agonists with the rank order of potency expected for beta ARs. There was an 89% reduction in catecholamine concentration in rat ventricles after 1 week of 6-hydroxydopamine treatment, before implantation of the minipumps. Chronic infusion of isoproterenol induced significant downregulation (63% to 74%) of beta 2ARs in atrial and ventricular myocytes, coronary arterioles, and connective tissue but no change in beta 1ARs in these regions in rats with intact sympathetic innervation. Similar changes were seen in denervated animals. There was a marked reduction in beta 2ARs but small insignificant decreases in beta 1ARs, despite the fact that in the denervated animals there was upregulation of beta 1ARs in atrial and ventricular myocytes (approximately 25%). Our study suggests that beta 1ARs in the heart are not significantly downregulated by chronic agonist exposure and that this is unrelated to sympathetic innervation. The underlying mechanism of preferential regulation of beta AR subtypes remains to be elucidated but may be related to differences in the molecular structure between beta 1ARs and beta 2ARs. PMID- 8403265 TI - Active oxygen species play a role in mediating platelet aggregation and cyclic flow variations in severely stenosed and endothelium-injured coronary arteries. AB - A canine model with cyclic flow variations (CFVs) in stenosed and endothelium injured coronary arteries was used to examine the role of active oxygen species in platelet aggregation in vivo. We studied 90 anesthetized dogs in which the pericardial cavity was opened and the heart was exposed. The velocity of blood flow in the left anterior descending coronary artery (LAD) was monitored by a pulsed Doppler flow probe. In 67 dogs, the LADs were stenosed by applying external constrictors at the site where the endothelium was mechanically injured. CFVs developed in all 67 dogs. Treatment with the antioxidants recombinant human copper-zinc superoxide dismutase (r-h-CuZnSOD), recombinant human manganese superoxide dismutase (r-h-MnSOD), and catalase eliminated platelet aggregation associated coronary CFVs in 63%, 62%, and 64% of animals, respectively. Intravenous infusion of epinephrine restored CFVs in most dogs. Ketanserin, a serotonin (5-hydroxytryptamine2) receptor antagonist, abolished epinephrine restored CFVs and eliminated CFVs in dogs in which CFVs had not been eliminated by free radical scavengers. In an additional 23 dogs, the LADs were stenosed but not mechanically injured. For control studies, saline was infused into the LADs of 5 dogs. Xanthine/xanthine oxidase was infused into the LADs of 8 dogs and induced CFVs in 4. Hydrogen peroxide was infused into the other 10 dogs and induced CFVs in 9. Histological analysis of the coronary artery revealed that the intima was significantly injured by the infusion. In ex vivo platelet aggregation studies, the in vivo treatment with r-h-CuZnSOD, r-h-MnSOD, and catalase significantly inhibited platelet aggregation induced by platelet-activating factor. Thus, active oxygen species are involved in mediating platelet aggregation and cyclic flow variations in stenosed and endothelium-injured canine coronary arteries in vivo. PMID- 8403266 TI - Expression of a minimal K+ channel protein in mammalian cells and immunolocalization in guinea pig heart. AB - The minimal K+ channel protein (minK, also called IsK) is structurally dissimilar to other cloned voltage-gated ion channels. minK is a 15-kD polypeptide with only one potential transmembrane helix. Published data suggest that the current associated with minK expression in Xenopus oocytes may be related to the slow cardiac delayed rectifier K+ current (IKs). However, the fact that minK expression has been limited exclusively to Xenopus oocytes has caused continuing concern about the nature of this protein and its molecular link to known mammalian K+ channels. We report in the present study the first expression of minK activity in transiently transfected mammalian (HEK 293) cells and demonstrate that the characteristics of the expressed minK current are similar to those of IKs recorded from guinea pig heart cells under similar experimental conditions. We also show that an antibody directed against the minK channel protein reacts with a surface antigen on adult guinea pig ventricular myocytes and sinoatrial nodal cells, where IKs is the dominant outward K+ current. The data provide strong evidence that a minK-like protein underlies IKs. PMID- 8403267 TI - Subunit-dependent modulation of recombinant L-type calcium channels. Molecular basis for dihydropyridine tissue selectivity. AB - At least four calcium channel subtypes (P, T, N, and L) have now been classified on the basis of their biophysical and/or pharmacological properties. L-type channels, a channel family particularly important to physiological function of the cardiovascular system, are identified by their slow voltage- and calcium dependent inactivation as well as their sensitivity to dihydropyridine (DHP) calcium channel antagonists. In this study, we report the results of experiments in which we have measured the DHP modulation of recombinant calcium channel activity in cells transfected with alpha 1 subunits of cardiac and smooth muscle L-type calcium channels. We find subunit-dependent differences in the voltage and concentration dependence of channel modulation. Our results provide evidence for a molecular basis for DHP sensitivity of heart and smooth muscle calcium channels and, additionally, indicate that, even within one family of calcium channels, slight differences in channel structure can cause marked differences in channel pharmacology. PMID- 8403268 TI - [Differentiation of schistosome species and strains by DNA hybridization]. AB - Genomic DNA from Schistosoma mansoni and S. japonicum adult worms was hybridized to a 32p-labelled pSM 889 probe after cleavage by restriction endonuclease BglII, BamHI, XbaI and EcoRI, or to a 32p-labelled pSM 389 probe after cleavage by EcoRI. The resulting hybridization banding patterns were significantly different between the two species. Genomic DNA from S. japonicum adult worms of Hunan, Hubei, Jiangxi, Zhejiang and Yunnan isolates was hybridized to a 32p-labelled pSM 389 probe after cleavage by EcoRI. The major hybridization bands were identical while the minor bands were more or less different among the isolates. Of them, isolates Hunan, Hubei and Zhejiang have similar minor bands while isolates Jiangxi and Yunnan have minor bands significantly different from those of the above three isolates and also markedly distinct from those of each other. These results indicate that the major hybridization banding patterns are unique to schistosome species while the minor banding patterns may serve as the basis for strain differentiation. PMID- 8403269 TI - [Screening antihydatid drugs using cultivated germinal cells of Echinococcus granulosus]. AB - Germinal cells isolated from Echinococcus granulosus cysts harbored in mice have been maintained in an in vitro culture system containing RPMI 1640 supplemented by 20% calf serum, and used as a model for screening anti-hydatid drugs. When the germinal cells were maintained in the medium for 6 days, the cell proliferation rate was rather high in the first four days but declined in the last two days. In screening drugs, 1.4 x 10(6) germinal cells were exposed to known effective drugs against metacestodes of E. granulosus in mice, such as mebendazole (Meb), albendazole (Alb) or praziquantel (Pra) at various concentrations. One to three days after exposure, cell counts were made daily in 3 samples of each drug concentration. The mean cell number of each group was compared with that of the control and the inhibition rate of the cell was then calculated. The results showed that the minimal effective concentrations of Meb, Alb and Pra, were 1.0 (48 h), 2.5 (24 h) and 10.0 (72 h) micrograms/ml, respectively, while the inhibition rates of the cell were 34.1, 55.7 and 18.5%. Interestingly, the in vitro effects of Meb, Alb and Pra were consistent to those obtained from the in vivo tests, ie Meb > Alb > Pra. Nevertheless, after exposure of germinal cells to Meb at 2.5 micrograms/ml for 24 h, the cells appeared in roughness, indistinction, shrunk or swelling, collapse, deformation and hole-like feature detected by light microscopy and scanning electron-microscopy, while the ultrastructure alterations of the cells noted by transmission electron-microscopy were lysis in cytoplasm, disruption or disappearance of nucleus and even darkness of the whole cell.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403270 TI - [Variations of free amino acids in hemolymph of Culex pipiens pallens during overwintering period]. AB - An analysis of the amino acids present in the hemolymph of Culex pipiens pallens caught from resting or overwintering places (blood meal: Sella 1, i.e. empty; ovary stage: under Christophers IIb) in Zhengzhou (34 degrees 43'N, 113 degrees 39E) indicated that 19, 18 and 18 kinds of free amino acids (FAAs) were detected during stages of preoverwintering, overwintering and postoverwintering respectively. Comparison of hemolymph amino acids between overwintering and preoverwintering stages revealed that the concentrations of the total FAAs and FAAs reduced significantly in overwintering except cystine and proline. The concentrations of cystine and proline during preoverwintering were 0.1563 +/- 0.0021 nmol/microliter and 22.2467 +/- 1.3537 nmol/microliter, but 0.2067 +/- 0.0058 nmol/microliter and 36.5467 +/- 0.4654 nmol/microliters during overwintering respectively. Comparison of hemolymph amino acids between postoverwintering and overwintering stages showed that the concentrations of all FAAs and total FAAs increased significantly during postoverwintering stage, and the concentrations of aspartic acid, glumatic acid, alanine, cystine, proline and total FAAs even exceeded those during preoverwintering stage respectively. PMID- 8403271 TI - [Long-term surveillance after basic elimination of bancroftian filariasis]. AB - The longitudinal and cross-sectional systemic surveillance have been conducted for 9-11 consecutive years in six counties (cities) of Guangxi Zhuang Autonomous Region after basic elimination of bancroftian filariasis. The two different control regimens had been used with DEC selective treatment followed by mass treatment of all persons and selective treatment followed by taking DEC medicated salt. During the former 6 years, residual microfilaremia cases could still be detected; whereas during the latter 5 years, no microfilaremia cases could be detected at all. The natural infection of vector mosquitoes showed negative. The positive rate of antibody in the populations was reduced to 1.4-5.5% detected by IFAT, reaching to the level of local non-endemic areas. The result indicated that the transmission of filariasis in these areas has been blocked. The authors suggested that a period of 10 years might be appropriate for surveillance after basic elimination of bancroftian filariasis. PMID- 8403272 TI - [Ultrastructural localization of 185 kDa and 82/41 kDa protective antigens in Plasmodium falciparum, FCC1/HN]. AB - Plasmodium falciparum FCC1/HN-infected human erythrocytes were embedded with LR White resin at low temperature. The 185 kDa and 82/41 kDa proteins in erythrocytic stages of P. falciparum were then immunolabeled by using the protective monoclonal antibodies (McAb)F6-D3 and F6-C2 with protein A-colloidal gold probe. The electron-microscopical observation showed that the 185kDa protein recognized by McAb F6-D3 was located on the surface of free and intracellular merozoites as well as the cytoplasm, plasma membrane, and parasitophorous vacuole membrane of immature schizonts. The 82/41 kDa proteins identified by McAb F6-C2 was located within the rhoptries of immature schizonts and mature merozoites. These results demonstrated ultrastructurally that the 185 kDa and 82/41 kDa protective antigens were merozoite surface antigen and merozoite rhoptry antigens of P. falciparum FCC1/HN, respectively. PMID- 8403273 TI - [Early ultrastructural evolution of murine malaria merozoites after entering red cells]. AB - A TEM study of murine malaria parasites, Plasmodium berghei and P. yoelii was performed by consecutive sampling in vivo to look into the early sequential changes in the ultrastructure of the merozoites after entering red cells. The results showed that once finishing invasion, the merozoite resided in the peripheral cytoplasm of the red cell, creating a bulge at the invasion site, with an additional unit membrane around it (parasitophorous vacuole); apical structures disappeared; the spherical body was degenerative or atrophic and separated from the mitochondrion and nucleus. The mitochondrion became more extended and the nucleus elongated and curved. There were more Er vesicles in the cytoplasm, taking a dilated polyangular shape. The inner double membrane was separated from the outer membrane and got into incomplete, winding, finally disappeared. Sometimes multimembranous bodies could be seen in the peripheral spaces. Once the dedifferentiation process was over, the merozoite was transformed into an early trophozoite, with a single plasma membrane and decreased density. Individual large Er vesicle with acute angles was found in the cytoplasm, and small food pills appeared beneath the plasma membrane; then the shape of the parasite changed from a ball-like one to a pie-like one, gradually the flat cell body rolled up, with its edges met and fused, resulting in the formation of a large food vacuole, with digestive vacuoles and pigment granules around it. Thus, it grew into a middle-aged trophozoite. PMID- 8403274 TI - [Efficacy of ivermectin for control of microfilaremia recurring after treatment with diethylcarbamazine: immunological observation]. AB - We compared the effect of a single dose of ivermectin (100 micrograms/kg) with that of a standard course of diethylcarbamazine (DEC) (6 mg/kg) on several parameters of the host's antifilarial immune response in 60 patients with bancroftian filariasis enrolled in a double-blind drug trial. All participants had measurable serum levels of antifilarial antibodies and parasite antigens. Drug-induced clearance of microfilaremia was associated with a temporary increase in HC11 antigenemia and a decrease in serum levels of antibodies to soluble filarial antigens. Antigenemia progressively declined in patients who remained a microfilaremic after treatment, but declined and then rose in persons with recurrent microfilaremia. Treatment triggered a sustained increase in serum levels of IL-1, IL-6, TNF alpha and IFN gamma in all patients. Although Ivermectin and DEC are believed to exert their antiparasite activity via different mechanisms, the same pattern of serological changes was observed in patients treated with either drug. PMID- 8403275 TI - [Analysis of proteins and antigens in nine strains of Toxoplasma gondii by means of SDS-PAGE and immunoblotting]. AB - The proteins and antigens of nine strains of Toxoplasma gondii were analyzed by SDS-PAGE and immunoblotting. These strains including RH strain, Zs-2 strain, CN strain and SH1, SH2, SH3, SH4, SH5, SH8 strains, were obtained from other laboratories or isolated from deformity fetus in our laboratory. SDS-PAGE analysis revealed that these strains were very close in the major bands of proteins and differences were observed only in part. There were common components in antigens of all strains using IgG antibody prepared from high-titre rabbit antisera raised against RH strain of Toxoplasma. PMID- 8403276 TI - [Studies on the mechanism of the protective immunity against Schistosoma japonicum--cellular immune responses]. AB - Kinetics of cellular immune responses was studied in C57BL/6 mice immunized with irradiation-attenuated vaccine, cryopreserved and irradiated vaccine or frozen thawn vaccine. The results and analysis of correlation between cellular responses and resistance showed that cellular immune responses played an important role in the protective immunity against Schistosoma japonicum. PMID- 8403277 TI - [Effect of gamma-irradiation on infectivity of Clonorchis sinensis metacercariae]. AB - The purpose of the present study was to observe the survival and development of Clonorchis sinensis metacercariae in their final hosts after Co-60 gamma irradiation exerting on both metacercariae isolated or in fish. Guinea pigs or albino rats were orally infected by gavage. Bio-assay, fecal examination for ova and dissection of infected animals were used for the estimation of minimum effective dose of gamma irradiation to control infectivity of Clonorchis sinensis metacercariae. Results showed that the minimum effective irradiation dose for isolated metacercariae was 0.05 kGy. The LD50 of the irradiation dose for metacercariae in fish was 0.05 kGy, and the minimum effective dose was 0.15 kGy. No significant difference in radiation susceptibility to Co-60 gamma irradiation was found among Clonorchis sinensis metacercariae in fishes collected at different localities in the northern, middle or southern parts of China. The present finding suggests that irradiation of the fish at a dose of 0.15 kGy could control the infectivity of Clonorchis sinensis metacercariae and thus be adopted as a control measure in preventive medicine. PMID- 8403278 TI - [Ultrastructural study on spermatogenesis and sustentacular cell of the testes of Schistosoma japonicum]. AB - This paper deals with the ultrastructural study of the spermatogenesis and sustentacular cell of the testes of Schistosoma japonicum by means of TEM. Albeit several papers have reported on the ultrastructural studies of the testes of male reproductive organ of S. mansoni, no systemic observation on the spermatogenesis of schistosome and ultrastructure of the testes of S. japonicum was available. The authors described the characteristics of the spermatogonium, spermatocyte and different appearance of spermatids. The latter was subdivided into 4 types, namely, Sda, Sdb, Sdc and Sdd, which could be identified by the features of nuclear shape and the density of matrix, the distribution of the mitochondria in the cytoplasm, the appearing and the formation of the acrosome-like structure, the presentation and elimination of the residual bodies, etc. The spermatids were embedded and interdigitated among the sustentacular cells, forming a complicated picture in the field of TEM. The acrosome-like structure and the inclusion of residual bodies were revealed in developing spermatids. The origin and the characteristics of these structures were discussed. PMID- 8403279 TI - [Comparative study on antigens of Cysticercus cellulosae by ELIB and ELISA]. AB - Urea-soluble antigens (Ag-u) and water-soluble antigens (Ag-w) of Cysticercus cellulosae were studied by using ELIB and ELISA, respectively. The results showed that using Ag-u and Ag-w, 9 and 14 bands of polypeptides could be recognized by sera from cysticercosis patients; 4 and 10 bands could be recognized by sera from hydatidosis patients; whereas 4 and 8 bands could be recognized by sera from clornochiasis patients. Using ELISA at serum dilutions of 1:400, the positive rate of sera against Ag-u and Ag-w was 96.6% and 86.6%, respectively in 60 cysticercosis patients; 16.6% and 46.6% respectively in 30 hydatidosis patients and negative in 24 clornorchiasis patients and 50 normal healthy persons, indicating that both Ag-u and Ag-w share common antigen components causing cross reaction with the sera from hydatidosis and clonorchiasis patients. However, such components were fewer in Ag-u than in Ag-w. In view of the fact that Ag-u is superior to Ag-w in both sensitivity and specificity, Ag-u might be better used for the diagnosis of cysticercosis. PMID- 8403280 TI - [Occurrence of Culex modestus inatomii Kamimura et Wada in Rongcheng city, Shandong Province]. AB - A mosquito survey was made at Rongcheng City, Shandong Province in August, 1991. Altogether 4 genera and 8 species of mosquitoes were collected in this survey, namely Aedes dorsalis (Meigen), Anopheles sinensis Wiedemann, An. yatsushiroensis Miyazaki, Culex fuscanus Wiedemann, Cx. modestus inatomii Kamimura et Wada, Cx. pipiens pallens Coquillett, Cx. tritaeniorhynchus Giles and Mansonia uniformis (Theobald). Among them Cx. modestus inatomii and An. yatsushiroensis were found to be the first record for China and Shandong Province respectively. The morphology and taxonomic status of the above two species are discussed briefly. PMID- 8403281 TI - [Detection of circulating antigen in urine from mice infected with Toxoplasma tachyzoites]. AB - Urine samples collected from three groups of mice infected experimentally with different numbers doses of Toxoplasma trophozoites were detected for the presence of Toxoplasma circulating antigen by using fast ELISA. The results showed that presence of circulating antigen in all three infected groups of mice in comparison with the normal control group. Toxoplasma circulating antigen was detected on days 5, 4 and 3 after infection in light-, moderate- and heavy infection groups, respectively. The concentration of circulating antigen was on a parallel with the duration of infection. Western blot analysis of the Toxoplasma circulating antigen in urine revealed the existence of seven specific bands with molecular weights of 75, 67, 55, 43, 30, 28 and 22 kDa. PMID- 8403282 TI - [ELIB of specific antigens in in vitro cultivated germinal cells of Echinococcus granulosus]. AB - Specific antigens are detected in cyst fluid and cyst wall of Echinococcus granulosus, as well as germinal cells cultivated in vitro by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme linked immunotransfer blot technique (ELIB), using the sera from mice infected with protoscoleces of E. granulosus for at least 10 months. A specific reaction band of 52 kDa or 38 kDa was detected in soluble protein of germinal cells and cyst fluid, respectively, but these two reaction bands were present concurrently in the cyst wall. The sera from 7 normal controls, 7 hydatidosis patients and 3 cysticercosis patients were used to study the specific antigen from the germinal cells. The results noted that the reaction band of 52 kDa was seen in all the sera from hydatidosis patients, while the sera from normal human controls and cysticercosis patients showed no reaction band. PMID- 8403283 TI - [Preliminary study on monoclonal antibodies against Cysticercus cellulosae]. AB - In order to produce valuable monoclonal antibodies for immunodiagnosis of cysticercosis, hybridoma technique was adopted using the fusion of splenic lymphocytes from BALB/c mice immunized with Cysticercus cellulosae antigens and the myeloma cell line NS-1. Through screening and cloning, three monoclonal antibody cell lines have been established. Immunoglobulin subclasses of 3 lines were determined to be IgG1. These monoclonal antibodies were found to be highly specific and without cross reaction with Echinococcus granulosus hydatid cyst and Cysticercus fasciolasis. Antigen fractions specifically reacting to monoclonal antibodies were demonstrated with Western blot method. The results showed that specific reaction bands of SCW located in 25 kDa and 17 kDa regions. PMID- 8403284 TI - [Effect of Schistosoma japonicum infection on liver drug-metabolizing enzymes of mice]. AB - In this paper, the effect of Schistosoma japonicum infection on liver drug metabolizing enzymes of mice was studied. The prolongation of hypnosis duration of sodium pentobarbital in vivo and the inhibition of liver aniline hydroxylase (AH), aminopyrine N-demethylase (APD) as well as cytochrome P-450 (Cyt P-450) in vitro were observed in mice infected with 20, 40 and 60 cercariae of S. japonicum after 6 weeks. Meanwhile, with 30 cercariae infection, no significant inhibition of the activity of AH, APD and Cyt P-450 was observed in mice 4 weeks after infection, but significant inhibition of these enzymes was shown by 8 weeks. As compared with control, the activity of AH, APD and Cyt P-450 decreased to 32.6%, 13.7% and 7.7% at 8 weeks. The results indicate that infection of S. japonicum can inhibit the liver drug-metabolizing capacity of mice, and the degree of inhibition depends on the infectivity and the duration of infection. PMID- 8403285 TI - Women in biomedical science. Through the looking glass. PMID- 8403286 TI - Pending legislation and tobacco industry deception. PMID- 8403287 TI - Transcatheter occlusion of patent ductus arteriosus with Gianturco coils. AB - BACKGROUND: Transcatheter occlusion with Gianturco coils has been attempted in a small number of patients with tiny (< or = 1.5-mm diameter) patent ductus arteriosus, and preliminary results have been encouraging. The present study extends this method to larger ductus sizes and makes recommendations for proper coil size selection. METHODS AND RESULTS: Coil occlusion was attempted in 24 consecutive patients with patent ductus arteriosus who did not require other cardiac surgery. Median patient age was 4.2 years (8 months to 30 years), and mean ductus diameter was 1.7 +/- 0.8 mm. Two instances of coil embolization occurred in the first 4 patients, with successful coil retrieval. Based on this experience, we proposed that the coil helical diameter should be twice or more the minimum ductus diameter, with coil length sufficient for three or more loops. With these recommendations, coils were successfully implanted in the subsequent 20 consecutive patients. Of the 22 patients with successful coil implantation, 15 (68%) had no residual shunting, and 7 had trace residual shunting by angiography. The continuous murmur was abolished in all 22 patients. No significant complications occurred, and all patients were discharged within 24 hours of successful coil implantation. No change in the systolic pressure gradient between main and left pulmonary artery or ascending and descending aorta was observed. CONCLUSIONS: Transcatheter occlusion of patent ductus arteriosus can be safely and effectively achieved in patients with ductus diameters up to 3.3 mm. Coil occlusion does not cause obstruction to flow in the left pulmonary artery or descending aorta. Coils should be selected to provide a helical diameter twice or more the minimum ductus diameter and a length sufficient for three or more loops. PMID- 8403288 TI - Lipids and lipoproteins predicting coronary heart disease mortality and morbidity in patients with non-insulin-dependent diabetes. AB - BACKGROUND: The aim of this study was to investigate the association of lipoprotein fractions with the future risk of coronary heart disease (CHD) in patients with non-insulin-dependent diabetes (NIDDM). METHODS AND RESULTS: At baseline, lipoprotein fractions were determined in 313 diabetic patients with NIDDM (153 men and 160 women), and these patients were followed up for 7 years with respect to CHD events (CHD death or all CHD events including CHD death or nonfatal myocardial infarction). Altogether, 56 NIDDM patients (28 men and 28 women) died from CHD and 25 had a nonfatal myocardial infarction (17 men and 8 women) during the follow-up. NIDDM patients having these CHD events during the follow-up had higher levels of total and very-low-density lipoprotein (VLDL) triglycerides and VLDL cholesterol and lower levels of high-density lipoprotein (HDL) and HDL2 cholesterol than those without CHD events. The risk for CHD death was fourfold and for all CHD events, twofold higher among diabetics with low HDL cholesterol (< 0.9 mmol/L) than among diabetics with HDL cholesterol > or = 0.9 mmol/L. High triglyceride level (> 2.3 mmol/L) was associated with a twofold increase in the risk of CHD events. In multiple logistic regression analyses, HDL was inversely associated with CHD events and VLDL triglycerides with CHD events in NIDDM patients with low HDL cholesterol level (< or = 1.12 mmol/L). CONCLUSIONS: Our 7-year follow-up study gives evidence that low HDL and HDL2 cholesterol, high VLDL cholesterol, and high total and VLDL triglycerides are powerful risk indicators for CHD events in patients with NIDDM: PMID- 8403289 TI - Insulin resistance is an important determinant of left ventricular mass in the obese. AB - BACKGROUND: Obesity in adults is associated with increased left ventricular (LV) mass. The mechanism for this is unclear, however. We tested the hypothesis that insulin resistance is an important independent contributing factor to LV mass in the healthy obese population. METHODS AND RESULTS: The study population consisted of 40 normotensive, nondiabetic, otherwise healthy obese subjects with body mass index (BMI) > 25 kg/m2. LV mass was echocardiographically determined according to the Penn convention, using the formula of Devereux and Reichek. Insulin resistance was assessed using indices derived from Intravenous Glucose Tolerance Test (IVGTT): insulin level at baseline, insulin level at 90 minutes of IVGTT (insulin-90), insulin integration over 90 minutes of IVGTT, and rate of glucose disposal (k value). Insulin-90 (r = .61, P = .0001), k value (r = .55, P = .003), insulin integration over 90 minutes (r = .46, P = .003), basal insulin (r = .44, P = .005), and BMI (r = .59, P = .0001) were all strongly correlated with LV mass by univariate analysis. No significant correlation was found with blood pressure or age. In multivariate regression analysis, only insulin-90 and k value correlated significantly with LV mass (P = .03, P = .02, respectively), accounting for 50% of the variance of LV mass, whereas the association with BMI became insignificant (P = .2). CONCLUSIONS: LV mass in the normotensive nondiabetic obese population is strongly associated with, and may be mediated by, the degree of insulin resistance and its associated hyperinsulinemia, independent of BMI and blood pressure. PMID- 8403290 TI - Height and incidence of cardiovascular disease in male physicians. AB - BACKGROUND: An inverse association between height and risk of coronary heart disease (CHD) has been reported in several case-control and cohort studies, but the reasons for the association remain uncertain. We evaluated this association among 22,071 male physicians, a population homogeneous for high educational attainment and socioeconomic status in adulthood. METHODS AND RESULTS: The study population was comprised of participants in the Physicians' Health Study, a randomized, double-blind, placebo-controlled trial of low-dose aspirin and beta carotene in the primary prevention of cardiovascular disease and cancer among US male physicians, aged 40 to 84 years, in 1982. Participants were classified into five height categories at study entry, from shortest to tallest, and were followed an average of 60.2 months to determine the incidence of myocardial infarction (MI), stroke, and death from cardiovascular disease. Men in the tallest (> or = 73 in. or 185.4 cm) compared with the shortest (< or = 67 in. or 170.2 cm) height category had a 35% lower risk of MI (relative risk, 0.65; 95% confidence interval, 0.44 to 0.99; P = .04), after adjusting for known cardiovascular risk factors. Further, a marginally significant inverse trend (P trend = .05) across the height categories was observed. Although the relationship was not strictly linear, for every inch of added height, there was an approximate 2% to 3% decline in risk of MI. In contrast, men in the tallest compared with the shortest height category had only small and nonsignificant decreases in risk of stroke and cardiovascular death. While no significant trend in risks of these end points across the height categories was observed, the numbers of events for these end points were far less than for MI, and thus the confidence intervals were wide. CONCLUSIONS: These data indicate that height is inversely associated with subsequent risk of MI. At this time, a few mechanisms are plausible, but none are convincing. Other epidemiological and basic research efforts are needed to explore a variety of physiological correlates of height that may be responsible for mediating the height-MI association. In the meantime, while height is not modifiable, it is easy to measure and may be useful to evaluate CHD disease risk profiles and target lifestyle interventions. PMID- 8403291 TI - Role of preclinical cardiovascular disease in the evolution from risk factor exposure to development of morbid events. AB - Conventional risk factors (especially high arterial pressure, elevated cholesterol and glucose levels, and cigarette smoking) are useful predictors of morbid atherosclerotic and hypertensive events, and their control variably reduces the incidence of events. However, both the ability to predict risk and the ability to reduce it by modification of established risk factors are limited. These limitations occur in part because the progression from risk factor exposure to morbid events depends on the variable likelihood that individuals exposed to the same risk factors will progress through two stages: the development of asymptomatic or "preclinical" anatomic and functional cardiovascular disease in response to standard risk factors and other variables, and the precipitation of morbid events by progression of preclinical disease or by the action of additional "triggering" mechanisms in the presence of preclinical disease. Advances in diagnostic methodology now make possible accurate noninvasive detection in many asymptomatic individuals of preclinical disease such as left ventricular hypertrophy, carotid atherosclerosis, and renal dysfunction. Progress in elucidating stimuli to left ventricular hypertrophy and systemic atherosclerosis suggests that focusing research separately on these two stages of disease evolution is a fruitful strategy. The closer relation of measures of preclinical disease than risk factors with the subsequent risk of complications indicates that their detection improves clinical risk stratification. However, critical testing of whether clinical outcome is improved or treatment cost is lowered by basing antihypertensive or antihyperlipidemic treatment decisions in part on the presence of preclinical cardiovascular disease is needed before this strategy is adopted on a widespread scale. PMID- 8403292 TI - Effects of age and aerobic capacity on arterial stiffness in healthy adults. AB - BACKGROUND: It has been well established that arterial stiffness, manifest as an increase in arterial pulse wave velocity or late systolic amplification of the carotid artery pressure pulse, increases with age. However, the populations studied in prior investigations were not rigorously screened to exclude clinical hypertension, occult coronary disease, or diabetes. Furthermore, it is unknown whether exercise capacity or chronic physical endurance training affects the age associated increase in arterial stiffness. METHODS AND RESULTS: Carotid arterial pressure pulse augmentation index (AGI), using applanation tonometry, and aortic pulse wave velocity (APWV) were measured in 146 male and female volunteers 21 to 96 years old from the Baltimore Longitudinal Study of Aging, who were rigorously screened to exclude clinical and occult cardiovascular disease. Aerobic capacity was determined in all individuals by measurement of maximal oxygen consumption (VO2max) during treadmill exercise. In this healthy, largely sedentary cohort, the arterial stiffness indexes AGI and APWV increased approximately fivefold and twofold, respectively, across the age span in both men and women, despite only a 14% increase in systolic blood pressure (SBP). These age-associated increases in AGI and APWV were of a similar magnitude to those in prior studies of less rigorously screened populations. Both AGI and APWV varied inversely with VO2max, and this relationship, at least for AGI, was independent of age. In endurance trained male athletes, 54 to 75 years old (VO2max = 44 +/- 3 mL.kg-1.min-1), the arterial stiffness indexes were significantly reduced relative to their sedentary age peers (AGI, 36% lower; APWV, 26% lower) despite similar blood pressures. CONCLUSIONS: Even in normotensive, rigorously screened volunteers in whom SBP increased an average of only 14% between ages 20 and 90 years, major age associated increases of arterial stiffness occur. Higher physical conditioning status, indexed by VO2max, was associated with reduced arterial stiffness, both within this predominantly sedentary population and in endurance trained older men relative to their less active age peers. These findings suggest that interventions to improve aerobic capacity may mitigate the stiffening of the arterial tree that accompanies normative aging. PMID- 8403293 TI - Thermolabile defect of methylenetetrahydrofolate reductase in coronary artery disease. AB - BACKGROUND: To determine whether or not a moderate genetic defect of homocysteine metabolism is associated with the development of coronary artery disease, we studied the prevalence of thermolabile methylenetetrahydrofolate reductase, which is probably the most common genetic defect of homocysteine metabolism. METHODS AND RESULTS: Three hundred thirty-nine subjects who underwent coronary angiography were classified into three groups: (1) patients with severe coronary artery stenosis (> or = 70% occlusion in one or more coronary arteries or > or = 50% occlusion in the left main coronary artery), (2) patients with mild to moderate coronary artery stenosis (< 70% occlusion in one or more coronary arteries or < 50% occlusion in the left main coronary artery), and (3) patients with non-coronary heart disease or noncardiac chest pain (nonstenotic coronary arteries). The thermolability of methylenetetrahydrofolate reductase was prospectively determined in all subjects. Plasma homocyst(e)ine levels were then measured in those with thermolabile methylenetetrahydrofolate reductase. The traditional risk factors for coronary artery disease were thereafter ascertained by chart review of all subjects. The prevalence of thermolabile methylenetetrahydrofolate reductase was 18.1% in group 1, 13.4% in group 2, and 7.9% in group 3. There was a significant difference between the prevalence of thermolabile methylenetetrahydrofolate reductase in groups 1 and 3 (P < .04). All individuals with thermolabile methylenetetrahydrofolate reductase irrespective of their clinical grouping had higher plasma homocyst(e)ine levels than normal (group 1, 14.86 +/- 5.85; group 2, 15.36 +/- 5.70; group 3, 13.39 +/- 3.80; normal, 8.50 +/- 2.8 nmol/mL). Nonetheless, there was no statistically significant difference in the plasma homocyst(e)ine concentrations of these patients with or without coronary artery stenosis. Using discriminant function analysis, thermolabile methylenetetrahydrofolate reductase was predictive of angiographically proven coronary artery stenosis. The traditional risk factors- age, sex, diabetes, smoking, hypercholesterolemia, and hypertension--were not significantly associated with the presence of thermolabile methylenetetrahydrofolate reductase. CONCLUSIONS: Thermolabile methylenetetrahydrofolate reductase is a risk factor for coronary artery disease and is unrelated to other risk factors. PMID- 8403294 TI - Prognosis of myocardial infarctions involving more than 40% of the left ventricle after acute reperfusion therapy. AB - BACKGROUND: Prior studies based on autopsy data suggest that infarction of more than 40% of the left ventricle necessitates cardiogenic shock and death. METHODS AND RESULTS: Technetium-99m Sestamibi tomography was used prospectively to measure infarct size at discharge in 166 patients with acute myocardial infarction. Patients with previous myocardial infarction or revascularization were excluded from the trial. Sixteen patients were identified with final infarct sizes > 40% of the left ventricle despite acute reperfusion therapy. These 16 patients (13 men) had a mean age of 63 +/- 10 years; 44% had a previous history of angina. Ten patients had emergent coronary angioplasty only (mean time to percutaneous transluminal coronary angioplasty [PTCA], 6.0 +/- 3.0 hours); 6 had thrombolysis (mean time to tissue plasminogen activator, 4.0 +/- 1.5 hours), of which 2 had rescue PTCA (5 and 3 hours from onset of pain). Of 15 patients who had angiograms after therapy, 15 had open infarct-related arteries. The left anterior descending artery was the infarct-related artery in 14 (9 proximal and 5 distal lesions). Half the patients had only single-vessel disease. Infarct size measured 50 +/- 7% of the left ventricle (range, 42% to 68%). Ejection fraction by radionuclide angiogram was 0.33 +/- 0.09 and 0.38 +/- 0.07 at discharge and 6 weeks, respectively. Hospital complications included shock (1 patient), pulmonary edema (2), angina (3), symptomatic nonsustained ventricular tachycardia (1), transient complete heart block (2), and transient bifascicular block (1). At follow-up (13 +/- 9 months), the patient with shock had died, but the remaining 15 patients were asymptomatic (1 had late PTCA for angina). CONCLUSIONS: In the interventional and thrombolytic era, patients with large residual myocardial infarctions can survive without heart failure. PMID- 8403295 TI - Streptokinase induces intravascular release of platelet-activating factor in patients with acute myocardial infarction and stimulates its synthesis by cultured human endothelial cells. AB - BACKGROUND: Reocclusion of a successfully recanalized infarct-related artery may account for failure of thrombolytic therapy. Evidence suggests that the intravascular activation of platelets may limit the response to this treatment. The aim of the present study was to investigate whether platelet-activating factor (PAF), an ether lipid mediator with multiple potent biological activities, is synthesized during therapy with thrombolytic agents. Two sets of experiments were performed: (1) we extracted and quantified PAF in blood of patients with acute myocardial infarction treated or untreated with streptokinase (SK), and (2) since the endothelium/platelet interaction is thought to be at the basis of vascular reocclusion, we studied whether cultured human endothelial cells synthesize PAF after stimulation with SK or plasmin. METHODS AND RESULTS: PAF was extracted from blood samples immediately after acidification to destroy the acid labile PAF-acetylhydrolase in 25 patients with acute myocardial infarction treated (group A, n = 14) and untreated (group B, n = 11) with intravenous infusion of SK. PAF was detected in 10 of 14 patients of group A and none of group B. PAF began to be detectable 60 to 90 minutes after SK infusion and disappeared from the circulation within 120 to 180 minutes. Percent variation of platelet count over basal values correlated negatively with the amount of PAF present in the circulation at 90 minutes (r = -.719; P < .001) and at 120 minutes (r = -.652; P < .001). Cultured human umbilical cord vein-derived endothelial cells (ECs) synthesized PAF in a dose-dependent manner in response to SK and plasmin, with a synthesis that peaked at 15 minutes and persisted up to 30 minutes for SK and 2 hours for plasmin. PAF extracted from blood samples or from ECs was quantified by bioassay performed after purification by thin-layer chromatography and high-performance liquid chromatography (HPLC). PAF-bioactive material was characterized as PAF with physicochemical and enzymatic treatments, HPLC-tandem mass spectrometry, and specific PAF-receptor antagonists. CONCLUSIONS: The observation that PAF was detectable in the blood of patients of group A only after treatment with SK and was not detectable in patients with a comparable infarct not treated with SK (group B) suggested that SK stimulates the synthesis of this mediator either directly or via plasmin generation. Indeed, cultured human ECs synthesize PAF after stimulation with both SK and plasmin. PAF production by ECs may promote platelet activation and interaction of these cells as well as of circulating leukocytes with endothelium. These events may limit the beneficial effects of thrombolytic therapy. PMID- 8403296 TI - Race and prognosis after myocardial infarction. Results of the thrombolysis in myocardial infarction (TIMI) phase II trial. AB - BACKGROUND: To better understand the role of race/ethnicity in survival after acute myocardial infarction, we compared clinical and laboratory data, response to thrombolytic therapy, and clinical outcome in 2885 patients participating in the Thrombolysis in Myocardial Infarction Phase II (TIMI II) Trial among three groups of patients (2564 whites, 174 blacks, and 147 Hispanics). METHODS AND RESULTS: Differences were found in baseline characteristics among the three groups including (1) age (mean age for whites, 57.2 years; blacks, 54.8 years; Hispanics, 52.8 years; P < .001), (2) sex (percentage of women for whites, 17.6; blacks, 28.7; Hispanics, 14.3; P < .001), and (3) risk factor prevalence: current smoking (percent for whites, 49.4; blacks, 62.1; Hispanics, 55.1; P < .003), history of hypertension (percent for whites, 36.6; blacks, 55.7; Hispanics, 39.5; P < .001), and diabetes mellitus (percent for whites, 11.9; blacks, 22.4; Hispanics, 19.7; P < .001). Changes in hemostatic factors 5 hours after infusion of recombinant tissue plasminogen activator (rt-PA) revealed a more profound fall in fibrinogen levels in black patients compared with the response seen in Hispanic or white patients (mean change in fibrinogen +/- SD, mg/dL: 151.3 +/- 107.4, 112.2 +/- 97.0, 109.4 +/- 98.6; P < .001, respectively) without more frequent infarct-related artery patency or hemorrhagic complications. Mortality was similar in the white, black, and Hispanic patients through the first year after adjustment for baseline variables. CONCLUSIONS: TIMI II data yield evidence that (1) corroborates published reports of a high prevalence of classic cardiovascular risk factors among minority patients with acute myocardial infarction, (2) there is a greater decrease in fibrinogen levels 5 hours after the start of rt-PA infusion among black patients than in white and Hispanic patients without evidence of more frequent infarct-related artery patency or hemorrhagic complications, and (3) thrombolytic therapy with appropriate supplemental measures is associated with comparable 1-year mortality in white, black, and Hispanic patients. PMID- 8403297 TI - Initial experience with a direct antithrombin, Hirulog, in unstable angina. Anticoagulant, antithrombotic, and clinical effects. AB - BACKGROUND: Currently available antithrombotic therapy for unstable angina is unwieldy and occasionally ineffective. This study was designed to investigate the potential of Hirulog, a new synthetic specific antithrombin agent, for the management of this condition. METHODS AND RESULTS: A total of 55 patients in the acute phase of unstable angina received intravenous Hirulog according to one of two protocols. In an acute dose-escalating study, 0.02, 0.05, 0.1, 0.25, and 0.5 mg.kg-1 x h-1, each for 30 minutes, were infused in 15 patients. Prolongation of activated partial thromboplastin time (aPTT) (r = .95), fibrinopeptide A inhibition (r = .96), and Hirulog plasma levels (r = .91) correlated closely with the dose infused, with significant changes compared with baseline appearing at doses of 0.25 mg.kg-1 x h-1 and higher. The purposes of the second protocol were to determine whether the anticoagulant and antithrombotic effects of the drug were sustained during a 72-hour infusion and to assess whether such treatment prevented the complications of unstable angina. Based on the initial study, we planned to give a dose of 0.25 mg.kg-1 x h-1 to each patient until 2 patients failed therapy, then successively higher doses until a 95% success rate was achieved or adverse effects intervened, increasing the dose after two failures had occurred at each level. Five patients received the 0.25-mg.kg-1 x h-1 dose and 14 the 0.5-mg.kg-1 x h-1 dose before two failures occurred. Failure was observed in only one of 21 patients at the dose of 1 mg.kg-1 x h-1. aPTT (+/- SEM) levels increased to 62 +/- 5, 76 +/- 2, and 98 +/- 3 seconds at the three doses, with minimal intraindividual variation, and Hirulog plasma levels to 1050, 2100, and 4200 mg/mL, respectively. Fibrinopeptide A plasma levels decreased at all doses but more consistently at the dose of 1 mg.kg-1 x h-1. The overall clinical success rate was 87.5%: 60% (3/5) at the low dose, 86% (12/14) at the intermediate dose, and 95% (20/21) at the high dose. No deaths, myocardial infarctions, or bleeding complications occurred. CONCLUSIONS: In unstable angina patients, Hirulog infusions quickly and reproducibly yield stable, dose-dependent anticoagulant and antithrombotic effects with a favorable clinical efficacy profile. PMID- 8403298 TI - Effect of strenuous exercise on platelet activation state and reactivity. AB - BACKGROUND: It has been hypothesized that platelets are activated, or made more activatible, by strenuous exercise and that these changes may play a role in the genesis of exercise-induced coronary ischemia. Previous studies have yielded conflicting results but have used assays (eg, platelet aggregation, plasma platelet factor 4, and plasma beta-thromboglobulin) that are subject to methodological problems. METHODS AND RESULTS: In the present study, a whole blood flow cytometric method was used to study the platelet activation state and reactivity of 12 physically active and 12 sedentary individuals before and after standardized treadmill exercise testing. The peptide gly-pro-arg-pro (GPRP) was included in this assay to prevent fibrin polymerization and platelet aggregation, thus allowing the measurement of the reactivity to thrombin of individual platelets in the physiological milieu of whole blood. A panel of fluorescent labeled monoclonal antibodies was used to monitor activation-dependent platelet surface changes: downregulation of glycoprotein (GP) Ib (6D1) and upregulation of GMP-140 (S12), the GPIIb-IIIa complex (PAC1), and GPIV (OKM5). In samples obtained before exercise, platelets not exposed to thrombin showed no evidence of in vitro activation. In the sedentary subjects, exercise caused a consistent and significant augmentation of the platelet activation state and reactivity as judged by the binding of 6D1 in the presence of thrombin 0.05 U/mL (P < .001), 0.005 U/mL (P = .001), and 0 U/mL (P = .004) and by the binding of OKM5 in the presence of thrombin 0.05 U/mL (P < .001), 0.005 U/mL (P = .029), and 0 U/mL (P = .035). Exercise increased the binding of PAC1 at only a single thrombin concentration (0.005 U/mL, P = .027) and did not alter the binding of S12 at any thrombin concentration. In contrast, in the physically active subjects, exercise failed to cause a consistent alteration in either platelet activation state or platelet reactivity. No significant differences were found between the 12 male and 12 female volunteers. CONCLUSIONS: Strenuous exercise in sedentary subjects but not physically active subjects resulted in both platelet activation and platelet hyperreactivity. These changes were more readily detected with monoclonal antibodies directed against GPIb (6D1) and, to a lesser extent, GPIV (OKM5) rather than those directed against the GPIIb-IIIa complex (PAC1) and GMP 140 (S12). Platelet activation by thrombin, generally regarded as the most physiologically important agonist, can be studied in whole blood in a clinical setting through the use of the peptide GPRP. PMID- 8403299 TI - MK-383 (L-700,462), a selective nonpeptide platelet glycoprotein IIb/IIIa antagonist, is active in man. AB - BACKGROUND: Fibrinogen-dependent cross-linking of glycoprotein (GP) IIb/IIIa on activated platelets is the final mechanism leading to platelet aggregation. Inhibition of this mechanism may result in a novel antithrombotic agent. We studied the activity of MK-383 (L-700,462), a new, nonpeptide GPIIb/IIIa antagonist, in vitro and in vivo, in man. METHODS AND RESULTS: MK-383, a nonpeptide tyrosine derivative, dose-dependently inhibited fibrinogen-dependent platelet aggregation, in vitro. Binding of 125I-labeled fibrinogen to activated platelets was prevented in a competitive manner with an IC50 of 10 +/- 4.2 nmol/L. The activity and tolerability of MK-383 were evaluated in a two-part double-blind, placebo-controlled, dose-escalation study in healthy male subjects using 1- and 4-hour intravenous infusions. Effects on ADP- and collagen-induced ex vivo platelet aggregation (APA or CPA) and template bleeding time (TBT) were evaluated. Twenty-four subjects participated in the 1-hour part. Six received placebo and 18 MK-383 in doses ranging from 0.05 to 0.40 microgram.kg-1 x min-1. MK-383 inhibited platelet aggregation and prolonged bleeding time in a dose dependent manner. APA and CPA were totally inhibited at the end of infusion of 0.4 microgram.kg-1 x min-1 and returned to 55% and 89% of baseline, respectively, at 3 hours after infusion. TBT was prolonged at this dose from 5.0 +/- 1.3 minutes predose to 22.7 +/- 6 minutes at the end of the infusion (P < .01) and was normalized by 3 hours after infusion. In the 4-hour infusion part, 15 subjects received MK-383 (0.1 to 0.2 microgram.kg-1 x min-1), and five received placebo. Complete inhibition of ex vivo platelet aggregation was seen at 0.15 and 0.2 microgram.kg-1 x min-1. At 0.2 microgram.kg-1 x min-1, TBT was prolonged from 4.4 +/- 1.2 to 23.9 +/- 4.3 minutes at the end of infusion (P < .01) and remained slightly prolonged 3 hours after infusion (7.2 +/- 1.8 minutes). No adverse effects were observed in any of the 33 subjects receiving MK-383. CONCLUSIONS: The results from this study indicate that MK-383 appears to be well tolerated and active in man. It is the first nonpeptide GPIIb/IIIa antagonist that can be used to investigate the antithrombotic potential of this new class of antiplatelet agents. PMID- 8403300 TI - Coronary vasomotor effects of serotonin in patients with angina. Relation to coronary stenosis morphology. AB - BACKGROUND: Previous experimental studies have shown that the effect of serotonin on a coronary stenosis depends on whether that stenosis is compliant or fixed. However, the relation between coronary stenosis morphology and the response to serotonin in patients with angina is not known. METHODS AND RESULTS: Using computerized quantitative coronary angiography, we studied the effects of intracoronary infusion of serotonin on 38 coronary stenoses of different morphologies (concentric, eccentric, complicated) in 11 patients with stable angina and 4 with variant angina. In response to the maximum infused concentration of serotonin, 100% of complicated stenoses and 50% of concentric stenoses constricted by > or = 20% (P < .05). The magnitude of constriction was greater at eccentric stenoses (32.08 +/- 4.1%) than concentric stenoses (15.68 +/ 2.8%, P < .05) and greater in complicated stenoses (57.69 +/- 7.6%, P < .05) than eccentric stenoses. At complicated stenoses, the constriction was greater (0.85 +/- 0.16 mm, P < .05) than at the adjacent reference segments (0.42 +/- 0.12 mm). It was similar to the reference segment for both concentric and eccentric stenoses. The constriction at the stenosis was greater for irregular (complicated) lesions than for smooth (concentric and eccentric) lesions in both patients with stable (51.8 +/- 7.3% versus 22.5 +/- 4.1%, P < .001) and those with variant (77 +/- 17% versus 28.2 +/- 8.1%, P < .05) angina. There was a weak correlation (r = .39) of magnitude of constriction with stenosis length but not with baseline stenosis severity (minimum diameter). CONCLUSIONS: In these patients, the magnitude of the vasoconstrictor response to serotonin at the site of an atheromatous coronary plaque depends on the morphological characteristics of the plaque and is more closely related to irregular contour than stenosis severity or length. This relation suggests that variations in receptor type or density or in the smooth muscle cell response to stimulation may determine the response to locally released serotonin in patients with coronary disease. PMID- 8403301 TI - Residual flow to the infarct zone as a determinant of infarct size after direct angioplasty. AB - BACKGROUND: In acute myocardial infarction, residual flow to the infarct zone either through antegrade flow in the infarct-related coronary artery or collateral flow from the non-infarct-related arteries is often present before reperfusion therapy. The purpose of this study was to assess the influence of antegrade flow in the infarct-related artery and/or collateral flow to the infarct zone before successful direct angioplasty on infarct size and myocardial salvage in patients with acute evolving myocardial infarction. METHODS AND RESULTS: Sixty patients with acute evolving myocardial infarction underwent direct successful angioplasty without prior thrombolytic therapy. The myocardium at risk of infarction, the final infarct size, and myocardial salvage were measured by tomographic perfusion imaging with 99mTc sestamibi. Antegrade flow in the infarct-related artery before intervention was graded according to the Thrombolysis in Myocardial Infarction (TIMI) study group classification. Collateral flow to the infarct zone before angioplasty was also graded (0 through 3, 0 being no collateral flow). The presence of even minimal antegrade flow before angioplasty (TIMI grade 1) in the infarct-related artery compared with absent flow was associated with a significant reduction in final infarct size (9 +/- 17% versus 23 +/- 19% of left ventricle, P = .02) and a significant increase in myocardial salvage (23 +/- 16% versus 14 +/- 13% of left ventricle, P = .05) after angioplasty. When antegrade flow in the infarct-related artery was absent before angioplasty, the presence of collateral flow before angioplasty resulted in a significantly smaller final infarct size (P = .01) and more myocardial salvage (P = .05) after angioplasty. Both antegrade infarct-related artery flow and collateral flow to the infarct zone had significant independent ability to predict infarct size after angioplasty. When collateral grade and TIMI grade were added to provide an estimate of residual flow, a model including residual flow, myocardium at risk, and the interaction of residual flow and infarct site explained 83% of the variability in infarct size after angioplasty. CONCLUSIONS: The presence of antegrade flow in the infarct-related artery and/or collateral flow to the infarct zone before direct angioplasty in acute evolving infarction results in a smaller infarct size after direct successful angioplasty. PMID- 8403302 TI - Redefining the treatment of peripheral artery disease. Role of percutaneous revascularization. PMID- 8403303 TI - Angiographic-pathologic correlations after elective percutaneous transluminal coronary angioplasty. AB - BACKGROUND: The local effect of coronary angioplasty is evaluated on the basis of postangioplasty angiograms. Smooth-walled dilation is considered to represent minimal or no injury, whereas intraluminal haziness corresponds with wall laceration. This study correlates the preangioplasty and postangioplasty angiograms with the histopathology of the target sites. METHODS AND RESULTS: The study includes 12 patients, each undergoing an elective procedure, and covers 19 angioplasty sites. Smooth-walled dilation and intraluminal haziness were not mutually exclusive. The angiograms were interpreted as smooth-walled dilation (n = 3), smooth-walled dilation with intraluminal haziness (n = 4), intraluminal and extraluminal haziness (n = 5), extraluminal dissection (n = 5), spiraltype dissection (n = 1), and aneurysm (n = 1). The histology of the arterial segments revealed wall laceration in all. Smooth-walled dilation without intraluminal haziness correlated with laceration limited to the intima in two, but with medial injury in one. Smooth-walled dilation with intraluminal haziness correlated with laceration limited to the intima in two and with medial injury in two. Intraluminal and extraluminal haziness corresponded with extensive laceration with deep involvement of the media in each. Extraluminal dissection correlated with a dissection along the shoulder area of the plaque, creating a broad-based flap. The spiral-type dissection corresponded with a true dissection into the plaque-free media. The aneurysm correlated with partial washout of an atherosclerotic plaque. CONCLUSIONS: The angiographic image of intraluminal and extraluminal haziness indicates extensive medial laceration. Smooth-walled dilation, with or without intraluminal haziness, is not a reliable indicator. The study emphasizes the need to reconsider the interpretations of postangioplasty coronary angiograms. PMID- 8403304 TI - Vascular complications after balloon and new device angioplasty. AB - BACKGROUND: Despite their potential advantages, new coronary angioplasty devices may be associated with more frequent vascular complications than noted after standard balloon angioplasty, theoretically due to the larger sheaths and prolonged periods of anticoagulation required by some of these devices. This study sought to identify the incidence, predictors, and clinical outcome of vascular complications after new device angioplasty. METHODS AND RESULTS: The clinical course of 1413 patients was reviewed after balloon or new device angioplasty. Vascular complications were defined as formation of a pseudoaneurysm, arteriovenous fistula, retroperitoneal hematoma, or groin hematoma associated with a > 15-point hematocrit drop or the need for surgical repair. Stepwise logistic regression was used to identify independent predictors for vascular complications. Vascular complications developed after 84 (5.9%) procedures; they occurred more frequently after intracoronary stenting (14.0%) and extraction atherectomy (12.5%) than after balloon angioplasty (3.2%) (odds ratios, 4.86; P < .001, and 4.26, P < .05, respectively). Independent predictors of vascular complications included the use of intraprocedural thrombolytic agents (P < .01), intracoronary stenting (P < .005), or extraction atherectomy (P < .05); high maximum creatinine level (P < .005); low nadir platelet count (P < .001); longer periods of excess anticoagulation (P < .05); and the need for repeat coronary angioplasty (P < .005). Vascular complications were not related to the size of the arterial sheath used. CONCLUSIONS: Vascular complications developed more frequently after new device angioplasty than after balloon angioplasty, with the risk for vascular complications directly related to the degree of periprocedural anticoagulation. PMID- 8403305 TI - Predicting early and intermediate-term outcome of coronary angioplasty in the elderly. AB - BACKGROUND: Although the technical success rate of coronary angioplasty in the elderly in high, very old patients have increased risk of procedure-related death and late recurrence of severe angina. We proposed to determine baseline variables that predict early and intermediate-term failure of percutaneous transluminal coronary angioplasty (PTCA) in patients more than 65 so we could effectively stratify risk. METHODS AND RESULTS: We studied 982 patients at least 65 years old who had urgent or elective PTCA (1980 through 1990). Follow-up (mean, 25 months) was obtained for all patients. Multiple baseline variables were analyzed with univariate and multivariate logistic regression to select independent ones to fit predictive models for in-hospital death or myocardial infarction (overall rate, 6.3%), total in-hospital adverse outcome (overall rate, 18.7%), cumulative survival free of myocardial infarction (overall rate, 15% at 3 years), and cumulative survival free of late nonfatal myocardial infarction, bypass surgery, repeat PTCA, or recurrent severe angina (overall rate, 47% at 3 years). The most heavily weighted parameter in the probability regression equation for each end point was the number of diseased coronary artery segments with at least 70% stenosis. Advanced age was less important. The number of concomitant medical illnesses was predictive of late outcome but not early in-hospital events. Lowest risk quintile versus highest risk quintile event rate was 2.9% versus 14% for acute myocardial infarction or death and 17.2% versus 29% for cumulative in hospital events. For posthospital events at 3 years' follow-up, lowest risk quintile death or myocardial infarction rate was 4% versus 33% for highest risk quintile. For cumulative late adverse events at 3 years, the event rate was 28% versus 63% for the highest risk quintile. CONCLUSIONS: These results stratify patients at high and low risk of early and intermediate-term success after PTCA and identify elderly patients in whom PTCA is most appropriate. PMID- 8403306 TI - Evaluation of ketanserin in the prevention of restenosis after percutaneous transluminal coronary angioplasty. A multicenter randomized double-blind placebo controlled trial. AB - BACKGROUND: Ketanserin is a serotonin S2-receptor antagonist that inhibits the platelet activation and vasoconstriction induced by serotonin and also inhibits the mitogenic effect of serotonin on vascular smooth muscle cells. METHODS AND RESULTS: We conducted a randomized, double blind, placebo-controlled trial to assess the effect of ketanserin in restenosis prevention after percutaneous transluminal coronary angioplasty (PTCA). Patients received either ketanserin (loading dose, 40 mg 1 hour before PTCA; maintenance dose, 40 mg bid for 6 months) or matched placebo. In addition, all patients received aspirin for 6 months. Coronary angiograms before PTCA, after PTCA, and at 6 months were quantitatively analyzed. Six hundred fifty-eight patients were entered into the intention-to-treat analysis. The primary clinical end point of the study was the occurrence between PTCA and 6 months of any one of the following: cardiac death, myocardial infarction, the need for repeat angioplasty, or bypass surgery. It also included the need for revascularization actuated by findings at 6-month follow-up angiography. The primary clinical end point was reached by 92 (28%) patients in the ketanserin group and 104 (32%) in the placebo group (RR, 0.89; 95% CI, 0.70, 1.13; P = .38). Quantitative angiography after PTCA and at follow up was available in 592 patients (ketanserin, 287; control, 305). The mean difference in minimal lumen diameter between post-PTCA and follow-up angiogram (primary angiographic end point) was 0.27 +/- 0.49 mm in the ketanserin group and 0.24 +/- 0.52 mm in the control group (difference, 0.03 mm; 95% CI, -0.05, 0.11; P = .50). CONCLUSIONS: Ketanserin at the dose administered in this trial failed to reduce the loss in minimal lumen diameter during follow-up after PTCA and did not significantly improve the clinical outcome. PMID- 8403307 TI - Hemodynamic and neurohormonal effects of the angiotensin II antagonist losartan in patients with congestive heart failure. AB - BACKGROUND: Losartan is a new specific angiotensin II receptor antagonist with no agonist properties that provides the opportunity to study the consequences of angiotensin II blockade. The objective of the present study was to evaluate the hemodynamic and neurohormonal response to losartan in patients with congestive heart failure. METHODS AND RESULTS: After baseline hemodynamic measurements using balloon-tipped pulmonary artery and radial arterial catheters, patients were randomized to receive a single dose of placebo or 5, 10, 25, 75, or 150 mg losartan in a double-blind, sequential fashion. Hemodynamic and neurohormonal parameters were then measured periodically for 24 hours. Losartan caused vasodilation in a dose-dependent manner. By the area-under-the-curve method, the reduction in the mean arterial pressure and systemic vascular resistance grew larger up to a dose of 25 mg, but the higher 75- and 150-mg doses did not produce additional vasodilation. In response to losartan, there were compensatory increases in both angiotensin II concentrations and in plasma renin activity, which were greatest at the highest doses. Aldosterone concentrations were significantly lowered with losartan. CONCLUSIONS: Blockade of the angiotensin II receptor with the antagonist losartan causes vasodilator and neurohormonal effects in patients with congestive heart failure. The lack of additional vasodilator response with doses of more than 25 mg suggests that neurohormonal activation might limit the efficacy of high dose of losartan. PMID- 8403308 TI - Abnormalities of sodium handling and of cardiovascular adaptations during high salt diet in patients with mild heart failure. AB - BACKGROUND: Sodium retention and hormonal activation are fundamental hallmarks in congestive heart failure. The present study was designed to assess the ability of patients with asymptomatic to mildly symptomatic heart failure and no signs or symptoms of congestion to excrete ingested sodium and to identify possible early abnormalities of hormonal and hemodynamic mechanisms related to sodium handling. METHODS AND RESULTS: The effects of a high salt diet (250 mEq/day for 6 days) on hemodynamics, salt-regulating hormones, and renal excretory response were investigated in a balanced study in 12 untreated patients with idiopathic or ischemic dilated cardiomyopathy and mild heart failure (NYHA class I-II, ejection fraction < 50%) (HF) and in 12 normal subjects, who had been previously maintained a 100 mEq/day NaCl diet. In normal subjects, high salt diet was associated with significant increases of echocardiographically measured left ventricular end-diastolic volume, ejection fraction, and stroke volume (all P < .001) and with a reduction of total peripheral resistance (P < .001). In addition, plasma atrial natriuretic factor (ANF) levels increased (P < .05), and plasma renin activity and aldosterone concentrations fell (both P < .001) in normals in response to salt excess. In HF patients, both left ventricular end diastolic and end-systolic volumes increased in response to high salt diet, whereas ejection fraction and stroke volume failed to increase, and total peripheral resistance did not change during high salt diet. In addition, plasma ANF levels did not rise in HF in response to salt loading, whereas plasma renin activity and aldosterone concentrations were as much suppressed as in normals. Although urinary sodium excretions were not significantly different in the two groups, there was a small but systematic reduction of daily sodium excretion in HF, which resulted in a significantly higher cumulative sodium balance in HF than in normals during the high salt diet period (P < .001). CONCLUSIONS: These results show a reduced ability to excrete a sodium load and early abnormalities of cardiac and hemodynamic adaptations to salt excess in patients with mild heart failure and no signs or symptoms of congestion. PMID- 8403309 TI - Erythrocyte ion fluxes in essential hypertensive patients with left ventricular hypertrophy. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is an independent risk factor for cardiovascular morbidity and mortality in essential hypertension (EH). Several hemodynamic and nonhemodynamic factors have been involved in the development of LVH in hypertension, including abnormalities in cellular ion mobilization. METHODS AND RESULTS: We measured different ion transport systems in erythrocytes from 50 patients with EH classified as having or not having LVH measured by M mode echocardiography. Thirty-two EH patients (64%) exhibited criteria of LVH, and 18 (36%) did not. When the two groups were compared, patients with LVH were older (44.7 +/- 7.4 versus 37.6 +/- 9.2 years; P < .01) and exhibited higher rates of erythrocyte Na(+)-H+ exchange (9.8 +/- 4.1 versus 7.1 +/- 2.6 mmol.[L.cells.h]-1; P < .05) and higher intraerythrocyte Na+ content (8.5 +/- 1.3 versus 7.5 +/- 0.8 mmol/L per cell; P < .01). Systolic and diastolic blood pressure values, as well as biochemical, hormonal, and other erythrocyte ion transport systems studied did not differ between EH with or without LVH. The results of a multiple linear regression analysis using left ventricular mass index (LVMI) as the dependent variable showed that Na(+)-H+ exchange and the maximal rate of the Na(+)-K(+)-Cl- cotransport were the only two independently significant parameters associated with an increased LVMI. CONCLUSIONS: The increased rate of the erythrocyte Na(+)-H+ exchange and the decreased maximal rate of the Na(+)-K(+)-Cl- cotransport system are both associated with the presence of LVH in EH patients. These abnormalities of ion transport pathways tend to increase the intracellular Na+ content and may be involved in the pathogenesis of LVH in EH. PMID- 8403310 TI - Ionic mechanisms of electronic inhibition and concealed conduction in rabbit atrioventricular nodal myocytes. AB - BACKGROUND: The term "concealed conduction" is used in electrocardiography when a proximal (atrial or ventricular) impulse penetrates the atrioventricular (AV) node but fails to traverse it completely. Its penetration into the node is inferred by its after effects on the propagation of succeeding impulses. Concealed AV nodal conduction is a well-established phenomenon, but its precise cellular and subcellular mechanisms are unknown. It has been suggested that concealed conduction results from a transient impairment of excitability caused by the subthreshold depolarization (ie, electrotonic inhibition) that is elicited downstream of the site of block. METHODS AND RESULTS: We studied the ionic mechanism of electrotonic inhibition and concealed conduction in single myocytes isolated from the rabbit AV node. Cells were paced using just-threshold current pulses delivered at a constant (basic) cycle length of 1 second. Appropriately timed interpolation of a conditioning pulse of depolarizing but subthreshold current led to failure of the subsequent, previously successful activation. The ability of the subthreshold response to inhibit subsequent excitation was increased when the interval between the conditioning and succeeding pulses was shortened, when the amplitude of the conditioning pulse was increased, or when the inward sodium current was blocked by superfusion with tetrodotoxin (30 microM). Voltage clamp analysis demonstrated that electrotonic inhibition results from partial inactivation of the transient calcium current (ICa.T). Similar results were obtained using a mathematical model (Hodgkin-Huxley type) of the AV nodal myocyte. Additional simulations in a linear array of AV nodal cells showed that when a premature impulse fails to traverse the AV node, the subthreshold depolarization elicited downstream of the site of block may lead to a transient reduction of excitability with consequent delay or block of the succeeding impulse. CONCLUSIONS: The overall data strongly suggest that some of the electrocardiographic manifestations of concealed AV conduction are the result of electrotonic inhibition of excitability secondary to a transient decrease in ICa.T. PMID- 8403311 TI - Identification of reentry circuit sites during catheter mapping and radiofrequency ablation of ventricular tachycardia late after myocardial infarction. AB - BACKGROUND: Ventricular tachycardia reentry circuits in chronic infarct scars can contain slow conduction zones, which are difficult to distinguish from bystander areas adjacent to the circuit during catheter mapping. This study developed criteria for identifying reentry circuit sites using computer simulations. These criteria then were tested during catheter mapping in humans to predict sites at which radiofrequency current application terminated ventricular tachycardia. METHODS AND RESULTS: In computer simulations, effects of single stimuli and stimulus trains at sites in and adjacent to reentry circuits were analyzed. Entrainment with concealed fusion, defined as ventricular tachycardia entrainment with no change in QRS morphology, could occur during stimulation in reentry circuit common pathways and adjacent bystander sites. Pacing at reentry circuit common pathway sites, the stimulus to QRS (S-QRS) interval equals the electrogram to QRS interval (EG-QRS) during tachycardia. The postpacing interval from the last stimulus to the following electrogram equals the tachycardia cycle length. Pacing at bystander sites the S-QRS exceeds the EG-QRS interval when the conduction time from the bystander site to the circuit is short but may be less than or equal to the EG-QRS interval when the conduction time to the circuit is long. The postpacing interval, however, always exceeds the tachycardia cycle length. When conduction in the circuit slows during pacing, the S-QRS and postpacing intervals increase and the slowest stimulus train most closely reflects conduction times during tachycardia. Endocardial catheter mapping and radiofrequency ablation were performed during 31 monomorphic ventricular tachycardias in 15 patients with drug refractory ventricular tachycardia late after myocardial infarction. During ventricular tachycardia, trains of electrical stimuli or scanning single stimuli were evaluated before application of radiofrequency current at the same site. Radiofrequency current terminated ventricular tachycardia at 24 of 241 sites (10%) in 12 of 15 patients (80%). Ventricular tachycardia termination occurred more frequently at sites with entrainment with concealed fusion (odds ratio, 3.4; 95% confidence interval [CI], 1.4 to 8.3), a postpacing interval approximating the ventricular tachycardia cycle length (odds ratio, 4.6; 95% CI, 1.6 to 12.9) and an S-QRS interval during entrainment of more than 60 milliseconds and less than 70% of the ventricular tachycardia cycle length (odds ratio, 4.9; 95% CI, 1.4 to 17.1). Ventricular tachycardia termination was also predicted by the presence of isolated diastolic potentials or continuous electrical activity (odds ratio, 5.2; 95% CI, 1.8 to 15.5), but these electrograms were infrequent (8% of all sites). Combinations of entrainment with concealed fusion, postpacing interval, S-QRS intervals, and isolated diastolic potentials or continuous electrical activity predicted a more than 35% incidence of ventricular tachycardia termination during radiofrequency current application versus a 4% incidence when none suggested that the site was in the reentry circuit. Analysis of the postpacing interval and S-QRS interval suggested that 25% of the sites with entrainment with concealed fusion were in bystander areas not within the reentry circuit. At restudy 5 to 7 days later, 6 patients had no monomorphic ventricular tachycardia inducible, and inducible ventricular tachycardias were modified in 4 patients. None of these 10 patients have suffered arrhythmia recurrences during a follow-up of 316 +/- 199 days, although 4 continue to receive previously ineffective medications. CONCLUSIONS: Regions giving rise to reentry after myocardial infarction are complex and can include bystander areas, slow conduction zones, and isthmuses for impulse propagation at which radiofrequency current lesions can interrupt reentry. PMID- 8403312 TI - Alterations of heart rate and of heart rate variability after radiofrequency catheter ablation of supraventricular tachycardia. Delineation of parasympathetic pathways in the human heart. AB - BACKGROUND: Persistent inappropriate sinus tachycardia has been reported as a complication after radiofrequency (RF) ablation of the fast atrioventricular (AV) nodal pathway. The purpose of this study was to evaluate the prevalence of this complication and its mechanism using heart rate variability analysis. METHODS AND RESULTS: Time and frequency domain analysis of heart rate was performed in the electrophysiology laboratory immediately before and immediately after RF ablation in 64 patients with supraventricular tachycardia. Ablation targets in these 64 patients included the fast AV nodal pathway (n = 3), the slow AV nodal pathway (n = 14), a posteroseptal accessory pathway (n = 23), and a left lateral accessory pathway (n = 24). A control group of 21 patients undergoing diagnostic study but not ablation underwent identical analysis immediately before and at the conclusion of their procedure. Patients undergoing ablation also had time and frequency domain analysis performed on ambulatory 24-hour Holter tapes recorded before ablation and at 1 day, 1 month, and 6 months after ablation. Compared with preablation values, time domain analysis immediately after ablation revealed a significant increase in mean heart rate and significant reductions in heart rate variability expressed as SD, MSSD, and PNN50 in patients undergoing AV nodal modification or posteroseptal accessory pathway ablation. Frequency domain analysis revealed marked attenuation of high frequency (0.15 to 0.40 Hz) components, indicating parasympathetic denervation. These acute changes were not seen after ablation of left lateral accessory pathways or after diagnostic study without ablation. Time and frequency domain analysis of 24-hour ambulatory Holter monitors performed serially after ablation revealed resolution of abnormalities of heart rate and of heart rate variability 1 to 6 months after ablation, with reappearance of the high frequency parasympathetic component suggestive of reinnervation. CONCLUSIONS: RF ablation in the anterior, mid, and posterior regions of the low interatrial septum may disrupt preganglionic or postganglionic parasympathetic fibers located in these regions that are destined to innervate the sinus node. Such fibers become more scarce along the left AV groove with increasing distance from the posteroseptal space. Parasympathetic denervation may be one mechanism for persistent inappropriate sinus tachycardia after RF ablation. PMID- 8403313 TI - Assessment of heart rate variability in hypertrophic cardiomyopathy. Association with clinical and prognostic features. AB - BACKGROUND: Altered vascular responses during exercise and disturbed responses to autonomic function testing have been documented in hypertrophic cardiomyopathy (HCM) and are associated with markers of an adverse prognosis. Reduced heart rate variability (HRV) and baroreflex sensitivity are predictors of increased risk of sudden death after myocardial infarction, but the value of these parameters in HCM is unknown. METHODS AND RESULTS: To determine the clinical significance of HRV and its relation to markers of electrical and hemodynamic instability in HCM, the 24-hour Holter recordings of 104 patients in sinus rhythm and off medication were analyzed. Five nonspectral measures of HRV were computed. The frequency components of HRV were calculated by fast Fourier transformation of the RR time intervals; the areas under the low (0.04 to 0.15 Hz) and high (0.15 to 0.4 Hz) frequency portions of the spectrum were measured as indices of autonomic and specific vagal influences on HRV, respectively. Spectral and nonspectral measures were compared with clinical, echo/Doppler, and Holter variables. ANCOVA was performed to allow for the effect of age on differences between variables. Spectral and nonspectral measures of HRV were correlated (r > .65; P < or = .001), indicating that the different time-domain and frequency parameters reflected similar measures of HRV. Global measures of HRV including the standard deviation of the mean of RR intervals (SDRR) and the standard deviation of 5 minute mean RR intervals (SDANN) were increased in patients with an adverse family history of HCM (173 +/- 67 vs 131 +/- 38 milliseconds, P = .001, and 158 +/- 66 vs 116 +/- 36 milliseconds, P = .004, respectively). In patients with exertional chest pain, global nonspectral measures were reduced compared with asymptomatic patients (118 +/- 31 vs 152 +/- 53 milliseconds, P = .006, and 105 +/- 30 vs 136 +/- 52 milliseconds, P = .014, respectively). Specific vagal influences on HRV including the proportion of RR intervals more than 50 milliseconds different (PNN50) and the high frequency peak on spectral analysis were less in patients with supraventricular arrhythmias on Holter monitoring (7.2 +/- 8 vs 16 +/- 13%, P = .012, and 21 +/- 10 vs 28 +/- 13 milliseconds, P = .048, respectively). Similarly, both global and specific vagal measures of HRV were less in the 27 patients with nonsustained ventricular tachycardia on Holter (PNN50, 7.7 +/- 9 vs 15 +/- 13 milliseconds, P = .048, and high frequency component, 19 +/- 9 vs 28 +/- 13 milliseconds, P = .05. During follow-up, 10 patients, 9 of whom were aged less than 33 years, experienced catastrophic events; 6 were resuscitated from ventricular fibrillation and 4 died suddenly. Indices of HRV were similar in these 10 patients to indices in the 94 survivors. CONCLUSIONS: Time-domain and spectral measures of HRV yield similar information about the specific autonomic influences on the heart. Global and specific vagal influences on HRV were reduced in patients with symptoms and arrhythmias and global HRV is increased in patients with an adverse family history of HCM, but these indices do not add to the predictive accuracy of established risk factors. PMID- 8403314 TI - Tetralogy of Fallot with pulmonary atresia. Rehabilitation of diminutive pulmonary arteries. AB - BACKGROUND: Patients with tetralogy of Fallot, pulmonary atresia, and diminutive pulmonary arteries are a high-risk group for whom there is no consensus on the correct approach to medical management. The purpose of this report is to review a 14-year experience in the treatment of these patients comparing management schemes. METHODS AND RESULTS: Between January 1978 and August 1988, 91 patients with tetralogy of Fallot and pulmonary atresia had an adequate evaluation of their pulmonary artery anatomy before any surgical management. Forty-eight of these patients had diminutive pulmonary arteries (38 to 104 mm2/m2) supplied by aortopulmonary collaterals and were managed in four different fashions. Of the 9 patients repaired primarily, 7 died early and the two survivors had poor hemodynamic outcome. Of the 9 patients conservatively managed with no intervention before 5 years of age, 4 died and only 1 had a satisfactory hemodynamic result after repair. Of 10 shunted patients, 3 died and 3 had satisfactory repairs. Since 1984, we have rehabilitated pulmonary arteries with (1) right ventricle to pulmonary artery surgical graft, (2) balloon dilation of residual pulmonary artery stenoses and embolization of collaterals, and (3) surgical closure of ventricular septal defect and repair of remaining obstructions. Of 20 patients so managed, 7 died after various stages, but 10 of 20 had complete repairs. All repaired patients with subsystemic right ventricular pressures had at least one successful pulmonary artery dilation. CONCLUSIONS: A combined catheter-surgery approach begun at an early age in patients with tetralogy of Fallot and pulmonary atresia with diminutive pulmonary arteries appears to enhance the chances of satisfactory complete repair. PMID- 8403315 TI - Determination of the most appropriate velocity threshold for applying hemispheric flow convergence equations to calculate flow rate: selected according to the transorifice pressure gradient. Digital computer analysis of the Doppler color flow convergence region. AB - BACKGROUND: While flow convergence methods have been promising for calculating volume flows from color Doppler images, it appears that the velocity threshold used and the transorifice pressure gradient dramatically influence the accuracy of application of the simple hemispheric flow convergence equation for calculation of flow rate. The present in vitro study was performed to determine whether the value of velocity threshold at which the shape of proximal isovelocity surface best fits given shape assumptions with different orifice sizes and flow rates is predictable as a function independent of orifice size from clinically measurable peak velocity or transorifice pressure gradient information. METHODS AND RESULTS: In an in vitro model built to facilitate ultrasound imaging, steady flow was driven through circular discrete orifices with diameters of 3.8, 5.5, and 10 mm. Flow rates ranged from 2.88 to 8.28 L/min with corresponding driving pressure gradients from 14 to 263 mm Hg. At each flow rate, Doppler color-encoded M-mode images through the center of the flow convergence region were obtained and transferred into the microcomputer (Macintosh IIci) in their original digital format. Then, the continuous wave Doppler traces of maximal velocity through the orifice were derived for the calculation of driving pressure gradient. Direct numerical spatial velocity measurements were obtained from the digital color encoded M-mode velocities with computer software. For each flow rate, we could calculate flow volume from any number of velocity distance combinations with a number of assumptions and use the results to assess expected flow convergence shape based on a priori knowledge of the progression from oblate hemispheroid to hemisphere to prolate hemispheroid changes observed previously. Our results showed that for a given ratio of calculated flow rate to actual flow rate (0.7 and 1), the velocity threshold that could be used for the calculation of flow rate with a hemispheric flow convergence equation correlated well with the pressure gradient for a given orifice size, and the differences in velocity threshold that could be used this way among different orifice sizes once they were adjusted for the covariate pressure gradients were not statistically significant (P = .79 for ratio = 0.7, and P = .81 for ratio = 1). CONCLUSIONS: Our present study provides an orifice size-independent quantitative method that can be used to select the most suitable velocity threshold for applying a simple hemispheric flow convergence equation based on clinically predictable pressure gradients ranging from 40 to 200 mm Hg, and it offers a correction factor that can be applied to the hemispheric flow convergence equation when the pressure gradient is less than 40 mm Hg. PMID- 8403316 TI - Immediate and one-year safety of intracoronary ultrasonic imaging. Evaluation with serial quantitative angiography. AB - BACKGROUND: Intracoronary ultrasound (ICUS) has the ability to quantitatively evaluate vessel wall morphology and is well suited for serial studies of coronary artery disease regression and progression. However, the potential risk for catheter-induced endothelial damage and accelerated atherosclerosis in instrumented vessels is a concern. The acute effects as well as the 1-year safety of ICUS regarding its impact on the atherosclerotic process were assessed. METHODS AND RESULTS: The acute studies include 240 intracoronary studies performed in 170 cardiac transplant recipients. Patients were systematically heparinized. Only vessels > or = 2 mm in diameter were visualized. Coronary arteries of 38 patients were measured by quantitative coronary angiography in matched angiograms at an interval of 1 year after the initial ICUS examination was performed to assess long-term effects. The angiographic measurements in the previously instrumented and noninstrumented vessels were compared. Forty-nine vessels that had been imaged (IM) in these 38 patients with a 5F ICUS catheter were compared with 61 vessels not previously imaged (NIM) in the same patients. Absolute and percentage change in angiographically measured mean vessel diameters in the ICUS imaged and nonimaged segments were compared. Despite pretreatment with nitroglycerin, 20 patients (8.3%) had angiographically evident coronary spasm. In all cases, this was reversed by giving nitroglycerin. One year after the original imaging study, no difference was noted between imaged and nonimaged vessels in change in absolute vessel diameter (IM, -0.11 +/- 0.28 mm vs NIM, 0.07 +/- 0.22 mm; P = .49) or in percentage change in diameter (IM, -5 +/- 11% vs NIM, -3 +/- 7%; P = .32). CONCLUSIONS: Intracoronary ultrasound in cardiac transplant recipients was associated with no clinical morbidity and a low incidence of vessel spasm in large and medium-size coronary arteries. It does not accelerate progression of angiographically quantifiable coronary artery disease. This study suggests that ICUS can be safely used even in coronary arteries not undergoing interventions. PMID- 8403317 TI - Three-dimensional echocardiography. In vivo validation for left ventricular volume and function. AB - BACKGROUND: Current two-dimensional quantitative echocardiographic methods of volume assessment require image acquisition from standardized scanning planes. Left ventricular volume and ejection fraction are then calculated by assuming ventricular symmetry and geometry. These assumptions may not be valid in distorted ventricles. Three-dimensional echocardiography can quantify left ventricular volume without the limitations imposed by the assumptions of two dimensional methods. We have developed a three-dimensional system that automatically integrates two-dimensional echocardiographic images and their positions in real time and calculates left ventricular volume directly from traced endocardial contours without geometric assumptions. METHODS AND RESULTS: To study the accuracy of this method in quantifying left ventricular volume and performance in vivo, a canine model was developed in which instantaneous left ventricular volume can be measured directly with an intracavitary balloon connected to an external column. Ten dogs were studied at 84 different cavity volumes (4 to 85 cm3) and in conditions of altered left ventricular shape produced by either coronary occlusion or right ventricular volume overload. To demonstrate clinical feasibility, 19 adult human subjects were then studied by this method for quantification of stroke volume. Left ventricular volume, stroke volume, and ejection fraction calculated by three-dimensional echocardiography correlated well with directly measured values (r = .98, .96, .96 for volume, stroke volume, and ejection fraction, respectively) and agreed closely with them (mean difference, -0.78 cm3, -0.60 cm3, -0.32%). In humans, there was a good correlation (r = .94, SEE = 4.29 cm3) and agreement (mean difference, -0.98 +/- 4.2 cm3) between three-dimensional echocardiography and Doppler-derived stroke volumes. CONCLUSIONS: Three-dimensional echocardiography allows accurate assessment of left ventricular volume and systolic function. PMID- 8403318 TI - Accuracy of electronic digital calipers compared with quantitative angiography in measuring coronary arterial diameter. AB - BACKGROUND: Quantitative angiography is the accepted method for measuring coronary luminal diameter. Electronic digital calipers have been used to assess arterial diameters in vasomotor function studies and after interventional procedures. However, careful validation of calipers against quantitative angiography has not been described. METHODS AND RESULTS: We used digital calipers and quantitative angiography to measure 517 arterial diameters (88 nonstenotic segments) in 24 transplant patients undergoing vasomotor function studies with acetylcholine and nitroglycerin, 20 stenoses in 14 patients with coronary artery disease, and 15 stenoses in 15 patients before and after excimer laser facilitated coronary angioplasty and at 6 months' follow-up. In nonstenotic arterial segments ranging in size from 0.6 to 3.5 mm, calipers overestimated diameters measured by quantitative angiography by 0.29 +/- 0.21 mm (mean +/- SD) (limits of agreement, -0.13 to 0.71 mm). However, when the vasomotor responses were expressed as percent diameter change, the two methods did not differ significantly (-1 +/- 10%; limits of agreement, -21% to 19%). In the 35 stenoses measured before intervention and 30 stenoses measured after intervention, calipers and quantitative angiography differed by 3 +/- 9% (limits of agreement, 15% to 21%) across a range of stenosis severity (11% to 80%). Repeat caliper measurements by the same observer of the percent diameter change in the transplant patients and the percent stenosis in the coronary artery disease patients led to standard deviations of the differences of 9.3% and 7.6%, respectively. Two different observers recorded percent diameter change and percent stenosis that differed with standard deviations of 9.6% and 7.8%, respectively. CONCLUSIONS: Quantitative angiography and electronic digital calipers produce similar relative changes in arterial diameters and percent stenosis in a broad range of severities. Digital calipers thus are a rapid and convenient alternative to computerized quantitative angiography in certain research studies and clinical practice of assessing stenosis severity. PMID- 8403319 TI - A simple ultrasound approach for detection of recurrent proximal-vein thrombosis. AB - BACKGROUND: The objective of this study was to develop a simple ultrasound method for measuring thrombus regression in patients with proximal deep-vein thrombosis (DVT) and to test its utility for the detection of DVT recurrence. METHODS AND RESULTS: The study comprised a cross-sectional survey and a prospective investigation (149 and 145 patients, respectively). In both phases, the normalization rate of a previously abnormal ultrasound test, applying the criterion of full compressibility of the common femoral and popliteal veins (C-US method), was assessed. In the prospective study, the vein diameter under maximum compression (thrombus thickness) was measured in the abnormal venous segments at scheduled times (1, 3, 6, and 12 months). In patients presenting with suspected DVT recurrence, the procedure was repeated and results were compared with those available from the previous examination. Noncompressibility of a previously normal(ized) venous segment and enlargement of thrombus thickness (> or = 2 mm) were considered diagnostic of proximal DVT recurrence. The diagnostic accuracy of the C-US method alone, as well as of the combined ultrasound methods (C-US + thrombus thickness), was assessed against contrast phlebography. C-US test normalization occurred in only 30% of patients within 1 year. A significant reduction of the thrombus mass (P < .0001) was recorded throughout the entire study period. However, a major decrease in thrombus mass (> 50%) was recorded within the first 3 months. Of 29 patients who developed a suspected recurrent DVT, phlebography confirmed diagnosis in 11. The C-US method alone showed an excellent accuracy (100%) but was applicable in only 6 patients (21%). Both the sensitivity and the specificity for proximal DVT recurrence of the combined ultrasound methods were 100% (95% confidence interval, 69% to 100% and 81% to 100%, respectively) and were applicable in all patients. CONCLUSIONS: The serial ultrasound measurement of thrombus mass after an acute episode of DVT may allow the correct identification of patients who develop a recurrent proximal-vein thrombosis. PMID- 8403320 TI - Thallium reinjection demonstrates viable myocardium in regions with reverse redistribution. AB - BACKGROUND: The clinical significance and pathophysiological mechanisms of reverse redistribution on stress-redistribution thallium-201 myocardial scintigraphy in patients with chronic coronary artery disease are unclear. Recent studies have shown that thallium-201 reinjection is a useful technique for the detection of myocardial viability in chronic coronary artery disease. In this investigation we determined whether thallium reinjection distinguishes viable from nonviable myocardium in regions with reverse redistribution. METHODS AND RESULTS: We studied 39 patients with chronic stable coronary artery disease (age, 60 +/- 10 years), all of whom demonstrated reverse redistribution on standard exercise-redistribution thallium single-photon emission computed tomography (SPECT). Reverse redistribution was defined as > or = 10% decrease in relative thallium-201 activity between stress and redistribution images and included either the worsening of a perfusion defect apparent on post-stress images or the appearance of a new defect on the redistribution images. Thallium reinjection was performed immediately after the 3- to 4-hour redistribution study. Of 39 regions with reverse redistribution, 32 (82%) showed enhanced thallium-201 activity (> or = 10% increase) after reinjection. In the other 7 regions (18%), the scintigraphic defect persisted after reinjection. Abnormal Q waves were present in only 8 of 32 (25%) regions with enhanced thallium-201 uptake after reinjection compared with 5 of 7 (71%) regions not responding to reinjection (P < .05). Akinetic or dyskinetic wall motion was present in 3 of 32 (9%) regions showing enhanced uptake after reinjection, in contrast with 5 of 7 (71%) regions not responding to reinjection (P < .01). Critically stenosed or totally occluded coronary arteries supplied 24 of 29 (83%) regions with enhanced thallium-201 uptake after reinjection but only 2 of 7 (28%) regions not showing a positive response to reinjection (P < .05). Collateral circulation was detected in 23 of 29 (79%) regions with a positive thallium reinjection effect but in only 1 of the other 7 regions (P < .01). Sixteen of the 39 patients also underwent positron emission tomography using 18F-fluorodeoxyglucose (FDG) to assess glucose utilization and H2(15)O to assess regional blood flow. The 14 regions with reverse redistribution that responded to reinjection with enhanced thallium uptake all showed either normal patterns of FDG uptake and flow or an ischemic pattern with increased FDG uptake relative to flow. Reduced FDG uptake and reduced flow values were seen in the two regions not responding to thallium reinjection. CONCLUSIONS: These observations indicate that reverse redistribution in chronic coronary artery disease usually reflects viable myocardium, critically dependent upon collateral circulation. PMID- 8403321 TI - Mitral valve repair in rheumatic disease. The flexible solution. AB - BACKGROUND: Mitral valve repair in rheumatic disease is technically more difficult, and there is little information on the long-term stability of this technique. METHODS AND RESULTS: From January 1975 to December 1990, 327 patients underwent mitral valve repair with the Duran flexible ring annuloplasty for rheumatic valve disease. Mean age was 45.4 +/- 12.6 years (range, 23 to 73 years). The techniques used for valve repair include a Duran flexible ring annuloplasty in all cases, commissurotomy in 272 (83.2%), papillary muscle splitting in 171 (52.3%), and subvalvular apparatus repair in 59 patients (18.0%). One hundred one patients required associated tricuspid valve surgery (30.8%). Hospital mortality was 3.36%, being lower for patients with isolated mitral valve repair (2.7%) than those with mitrotricuspid surgery (4.9%). Mean follow-up was 8.6 years (range, 1 to 17 years) and was 96.5% completed. Thirty four patients required reoperation for severe mitral insufficiency in 12, mitral restenosis in 18, and aortic valve disease in 4. The actuarial curve free from reoperation for mitral cause at 16 years is 89.9 +/- 3.2%. Late mortality occurred in 42 patients (13.2%). Actuarial survival curve at 16 years is 84.0 +/- 3.2% for isolated mitral valve repair and 64.6 +/- 6.7% for mitrotricuspid patients. CONCLUSIONS: Mitral valve reconstruction with Duran flexible ring annuloplasty in rheumatic valve disease entails a low hospital mortality with satisfactory long-term clinical results. PMID- 8403322 TI - Experimental atrial septal defect closure with a new, transcatheter, self centering device. AB - BACKGROUND: Despite two decades of research, a transcatheter atrial septal defect closure device is not available for clinical use. We have designed a new superelastic Nitinol-Dacron, double-disk, self-centering, atrial septal defect closure device and studied its efficacy in a canine model of atrial septal defects. METHODS AND RESULTS: Atrial septal defects were created surgically in 20 adult dogs using either a 7.5-mm or 10-mm punch. Percutaneous transcatheter closures were attempted using a new device. The device sizes used were 20 mm in 6 dogs, 22 mm in 9, and 25 mm in 5 (22.1 +/- 1.9 mm, mean +/- SD). The stretched atrial septal defect diameter was 10.5 +/- 1.3 mm, and the device to stretched atrial septal defect diameter ratio was 2.1 +/- 0.3. Closures were successful in 19 studies and unsuccessful in 1. Angiography showed a left-to-right shunt in all 20 dogs before closure. Immediately after closure (n = 19), there were no shunts in 17 and trivial shunts in 2. Six dogs were followed for a period of 4.7 +/- 3.0 months (range, 2 to 8 months). The trivial shunt present in 1 animal immediately after closure had closed by the time of the repeat study. Spontaneous embolization of the device was not seen during follow-up. A solitary wire fracture was found 8 months after closure in 1 device. Light microscopy at 8 weeks in 3 dogs showed the devices to be covered by smooth endocardium, enmeshed in mature collagen tissue, with a minimal mononuclear cell infiltration. Retrievability was assessed by deliberately embolizing 4 devices in 2 dogs into the right atrium (n = 1) and pulmonary artery (n = 3). All devices were successfully retrieved with a snare. CONCLUSIONS: This feasibility study demonstrates that this new self-centering atrial septal defect closure device has a number of design features that permit effective and safe closures in a canine model. These results support the investigation of this device in human clinical trials. PMID- 8403323 TI - Induction of right ventricular hypertrophy with obstructing balloon catheter. Nonsurgical ventricular preparation for the arterial switch operation in simple transposition. AB - BACKGROUND: Recently, a successful result with a rapid two-stage arterial switch operation (ASO) was reported for patients with transposition of the great arteries (TGA) with low left ventricular pressure. In this procedure, the interval between pulmonary arterial banding and ASO was approximately 1 week. This successful result indicates the possibility of a nonsurgical ventricular preparation procedure using an obstructing balloon catheter prior to ASO. METHODS AND RESULTS: A 5F atrioseptostomy catheter was inserted directly into the main pulmonary artery in six lambs aged 20 to 38 days. After the chest was closed, the balloon was inflated twice a day for a period of 2 to 2.5 hours. This procedure was performed for 4 consecutive days. After the final inflation, the ratio of right ventricular weight to total ventricular weight was compared with that in an age-matched control group. After the final inflation, the peak systolic right ventricular pressure and the percentage of peak systolic right ventricular to peak systolic aortic pressure rose to 85.6 +/- 4.7 mm Hg (mean +/- 1 SD) and 79.6 +/- 8.6%, respectively. The percentages of the right ventricular weight to the total ventricular weight were significantly higher after the balloon inflation than those in the control group in terms of wet heart weight (29.5 +/- 1.2% versus 23.0 +/- 1.0%; P < .0001) and dry heart weight (27.0 +/- 2.0% versus 21.0 +/- 1.1%; P < .0001). CONCLUSIONS: The myocardial mass in the right ventricle increased after 4 days of intermittently applied pressure overload. Nonsurgical preparation of the ventricle for ASO in TGA is feasible. PMID- 8403324 TI - Acute effect of percutaneous transvenous mitral commissurotomy on ventilatory and hemodynamic responses to exercise. Pathophysiological basis for early symptomatic improvement. AB - BACKGROUND: Improvement of exertional dyspnea occurs immediately after percutaneous transvenous mitral commissurotomy (PTMC), but the pathophysiological basis for this early symptomatic improvement has not been elucidated. METHODS AND RESULTS: Exercise hemodynamic measurement and exercise ventilatory measurement with arterial blood gas analysis were performed in 21 patients aged 50.4 +/- 9.5 years (mean +/- SD) with symptomatic mitral stenosis before and a few days after PTMC. Exercise ventilatory measurement were also performed in 14 normal control subjects aged 48.9 +/- 4.9 years. After PTMC, mitral valve area increased (from 1.0 +/- 0.3 to 1.7 +/- 0.3 cm2, P < .001), mean mitral gradient (from 12.2 +/- 5.2 to 5.2 +/- 2.2 mm Hg, P < .001), and mean left atrial pressure (from 18.7 +/- 6.1 to 12.1 +/- 4.0 mm Hg, P < .001) decreased. All patients experienced significant symptomatic improvement soon after PTMC. Comparison of hemodynamic parameters at the same ergometer work rate showed a significant decrease in pulmonary artery systolic pressure (from 77 +/- 18 to 67 +/- 14 mm Hg, P < .001) and diastolic pressure (from 36 +/- 10 to 28 +/- 7 mm Hg, P < .001) and a significant increase in cardiac output (from 6.4 +/- 1.4 to 8.1 +/- 1.9 L/min, P < .001). Despite the improvement in exercise hemodynamics and symptoms, exercise capacity determined by peak oxygen uptake (from 18.0 +/- 2.9 to 18.6 +/- 3.1 mL.kg-1 x min-1) and anaerobic threshold (from 11.7 +/- 2.4 to 12.0 +/- 2.4 mL.kg 1 x min-1) remained unchanged. Excessive exercise ventilation, as assessed by the slope of the regression line between expired minute ventilation and carbon dioxide output, decreased significantly from 37.2 +/- 6.7 to 33.9 +/- 5.8 (P < .001), but remained significantly higher than that in the normal subjects (27.9 +/- 3.6, P < .01). The ratio of total dead space to tidal volume and total dead space per breath during exercise decreased significantly after PTMC (P < .05). The change in excessive exercise ventilation after PTMC was correlated with the change in dead space to tidal volume ratio (r = .59). CONCLUSIONS: Significant relief of exertional dyspnea immediately after PTMC is not accompanied by an improvement in exercise capacity. A decrease in excessive ventilation due to a decrease in physiological dead space resulting from hemodynamic improvement partly contributes to the early relief of symptoms after PTMC. However, lung compliance, which was not measured in the present study, may have changed after PTMC. This change may also contribute to the symptomatic improvement. PMID- 8403325 TI - Ductus arteriosus. Advanced differentiation of smooth muscle cells demonstrated by myosin heavy chain isoform expression in rabbits. AB - BACKGROUND: The closure of the ductus arteriosus (DA) is one of the most striking cardiovascular events that occur at birth. It has been attributed to oxygenation and intrinsic prostaglandins. However, selective constriction of DA suggests the presence of highly specialized contractile mechanisms in DA. We previously reported that smooth muscle myosin heavy chain isoforms, SM1 and SM2, are molecular markers for smooth muscle differentiation because of their unique expression pattern during vascular development. SM1 and SM2 are generated from a single gene through developmentally regulated alternative RNA splicing; SM1 is expressed in almost all stages of differentiation of the vascular smooth muscles, but SM2 is found only after birth. METHODS AND RESULTS: Immunohistochemistry was performed to study the expression of the different types of myosin heavy chain isoforms in DA of fetal and neonatal rabbits. Electron microscopic examinations were also carried out to demonstrate ultrastructural characteristics of ductus muscles. We found that SM2 is expressed before birth in the medial layer of DA, indicating advanced differentiation of smooth muscle cells in DA. The exact location of immunoreactivity for SM2 was in the smooth muscle cell of the medial layer of DA. Immunoreactivity for SM1, however, was not different for DA and adjacent great arteries. Transmission electron microscopy demonstrated greater amounts of myofilaments in medial smooth muscles of DA than those of aorta. CONCLUSIONS: These results indicate that smooth muscles in DA are more differentiated than those in other arteries, which may be one of the cellular mechanisms responsible for the unique closure of DA at birth. PMID- 8403326 TI - Persistent T-wave changes after alteration of the ventricular activation sequence. New insights into cellular mechanisms of 'cardiac memory'. AB - BACKGROUND: "Cardiac memory" refers to persistent T-wave changes on ECG that follow resumption of sinus rhythm after a period of altered activation sequence. Previous studies demonstrated that cardiac memory in intact dogs was abolished by 4-aminopyridine (4-AP), which blocks both the transient outward potassium current, Ito, and IK. METHODS AND RESULTS: We used standard microelectrode techniques to study the mechanism for cardiac memory in canine ventricular subepicardial and subendocardial slabs measuring 15 x 30 x 1 to 2 mm. Bipolar electrodes were used to stimulate slabs parallel to fiber axis, simulating normal activation, and perpendicular to fiber axis, simulating ventricular pacing. Four 30-minute periods of normal activation at a basic cycle length of 650 milliseconds were interrupted by three 20-minute periods of ventricular pacing at a basic cycle length of 450 milliseconds. We first recorded action potentials differentially from epicardial and endocardial slabs. The stimulation protocol induced changes in the "T" wave of the difference signals that mimicked cardiac memory and that could be explained on the basis of the transmural gradient in repolarization between epicardium and endocardium. This result was not obtainable with slow and rapid pacing from one site only. In subsequent experiments, action potential characteristics of epicardial and endocardial slabs were studied by the same pacing protocol with alternation between simulated normal activation and ventricular pacing. During ventricular pacing, the epicardial phase 1 notch and plateau amplitude decreased compared with normal activation. 4-AP (3 mmol/L) decreased notch size and plateau amplitude during normal activation in epicardium but not endocardium. In contrast, the local anesthetic lidocaine did not change notch size or plateau amplitude in epicardium or endocardium. CONCLUSIONS: These results suggest that the contribution to repolarization of specific potassium channels influences the memory phenomenon and that by blocking Ito and reducing the transmural voltage gradient for repolarization, 4-AP abolishes cardiac memory. PMID- 8403327 TI - Inhalation of sidestream cigarette smoke accelerates development of arteriosclerotic plaques. AB - BACKGROUND: Environmental tobacco smoke has been blamed for approximately 40,000 excess deaths from heart disease annually in the United States. As yet, no pathophysiological process that could be responsible for these deaths has been identified. Environmental tobacco smoke is composed mainly of aged and diluted sidestream smoke but also contains 15% to 20% exhaled mainstream smoke. Carcinogens, including nitrosamines and polynuclear aromatic hydrocarbons, are present in mainstream smoke and sidestream smoke. Carcinogen levels in sidestream smoke, unlike those in mainstream smoke, are not reduced in filtered cigarettes. The US Environmental Protection Agency has designated environmental tobacco smoke as a human (class A) carcinogen. In cockerels, subtumorigenic doses of polynuclear aromatic hydrocarbons carcinogens accelerate aortic arteriosclerotic plaque development. METHODS AND RESULTS: To determine whether sidestream smoke inhalation affects arteriosclerotic plaque development, we exposed cockerels to sidestream smoke (n = 30) or to filtered air (n = 12) in inhalation chambers for 6 hours per day, 5 days a week from 6 to 22 weeks of age (0.4% of projected lifespan). Chamber levels of carbon monoxide, total suspended particulates, and nicotine were measured regularly during the exposures. The abdominal aorta from each cockerel was cut into 10 segments, and the plaque index (mean plaque cross sectional area [mm2]/mean luminal circumference [mm] x 100) was calculated for each segment. There were no differences in plaque incidence or distribution between sidestream smoke-exposed and control cockerels; however, plaque indexes were significantly greater for sidestream smoke-exposed than control cockerels in all segments. CONCLUSIONS: Thus, relatively brief exposures to sidestream smoke early in life are sufficient to enhance arteriosclerotic plaque development. PMID- 8403328 TI - Cellular electrophysiological effects of hyperthermia on isolated guinea pig papillary muscle. Implications for catheter ablation. AB - BACKGROUND: The primary mechanism of tissue injury by radiofrequency catheter ablation is presumed to be thermally mediated. However, the myocardial cellular electrophysiological effects of hyperthermia are not well characterized. We used an in vitro model of isolated guinea pig right ventricular papillary muscle to investigate the acute cellular electrophysiological effects of hyperthermia. METHODS AND RESULTS: Excised guinea pig right ventricular papillary muscles were pinned in a high-flow tissue bath and superfused with Tyrode's solution at 37.0 +/- 0.5 degrees C. The superfusate temperature was rapidly changed to 38.0 to 56.0 degrees C for 60 seconds and then returned to 37.0 degrees C. Conventional microelectrodes were used to measure membrane potential (Vm), maximum rate of rise of the action potential (dV/dtmax), and action potential (AP) amplitude and AP duration at 50% (APD50) and 90% (APD90) repolarization. Hyperthermia resulted in (1) a progressive depolarization of Vm at temperatures > or = 40.0 degrees C, which became more prominent at temperatures > or = 45.0 degrees C; (2) changes in the AP characterized by a temperature-dependent increase in dV/dtmax and a temperature-dependent decrease in AP amplitude, APD50, and APD90; (3) reversible loss of cellular excitability within a temperature range of 42.7 to 51.3 degrees C (median, 48.0 degrees C); (4) irreversible loss of cellular excitability and tissue injury at temperatures > or = 50.0 degrees C; and (5) the development of abnormal automaticity at temperatures > 45.0 degrees C. CONCLUSIONS: Hyperthermia causes significant changes in myocardial cellular electrophysiological properties that include membrane depolarization, reversible and irreversible loss of excitability, and abnormal automaticity. There appear to be specific temperature ranges for reversible and irreversible electrophysiological changes. These observations may have important implications for tissue temperature monitoring during radiofrequency catheter ablation. PMID- 8403329 TI - Electrophysiological mechanisms in a canine model of erythromycin-associated long QT syndrome. AB - BACKGROUND: Erythromycin is known to prolong ventricular repolarization and has been associated with the occurrence of torsades de pointes. In this study, we have investigated potential mechanisms in vivo and in vitro for induction of an acquired long QT syndrome by erythromycin. METHODS AND RESULTS: Ventricular electrograms and endocardial monophasic action potentials were recorded in anesthetized open-chest dogs before and after administration of 40 to 120 mg/kg of erythromycin lactobionate. Conventional microelectrode techniques were used to record transmembrane action potentials in isolated dog Purkinje fibers and papillary muscles. Erythromycin at concentrations > 20 mg/L prolonged action potential duration. At higher concentrations (100 to 200 mg/L), erythromycin induced phase 2 and phase 3 early afterdepolarizations (EADs) both in vivo and in vitro. The effects of erythromycin on repolarization were more marked in Purkinje fibers than in papillary muscle. Pretreatment of Purkinje fibers with erythromycin antagonized the effects of dofetilide, a selective delayed-rectifier potassium channel (IK) blocker. Pretreatment with prazosin or tetrodotoxin had no effect on erythromycin-induced changes in action potential duration. CONCLUSIONS: These pharmacological studies suggest that erythromycin prolongs repolarization to a large extent by block of IK. In turn, prolongation of action potential duration resulting from erythromycin's actions on IK may promote the development of EADs. The induction of ventricular arrhythmias observed clinically after exposure to erythromycin may be related to the development of EADs. The rarity of occurrence of ventricular arrhythmias suggests that other predisposing factors contribute to the acquired long QT syndrome associated with erythromycin. PMID- 8403330 TI - Role of ATP-sensitive K+ channel on ECG ST segment elevation during a bout of myocardial ischemia. A study on epicardial mapping in dogs. AB - BACKGROUND: ATP-sensitive K+ channels are activated when the myocardium becomes ischemic. However, the role of the ATP-sensitive K+ current in the emergence of ECG ST changes during ischemia remained unclarified. METHODS AND RESULTS: The left anterior descending coronary artery (LAD) was cannulated and perfused with arterial blood from the carotid artery through a bypass tube in 8 anesthetized, open-chest dogs. An array of 60 unipolar electrodes mounted on a sock was used to record epicardial electrograms of the whole heart. Pinacidil (10 micrograms.kg-1 x min-1), an ATP-sensitive K+ channel opener, was infused into the bypass tube for 2 minutes, and the electrograms were recorded before and after the infusion. The elevation of the ST segment and the increase of QRST area were observed spatially over the LAD-perfused region. At the electrode showing the largest ST segment elevation, the activation recovery interval, an index of action potential duration, was shortened from 202 +/- 9 to 111 +/- 18 milliseconds (P < .001). These electrographic changes were similar to those noted in 2-minute coronary occlusion (n = 8). The extent of ST segment elevation during coronary occlusion was attenuated after the intravenous pretreatment with glibenclamide (0.3 mg/kg), a blocker of the KATP channel (n = 5). CONCLUSIONS: The findings of this study suggest that the activation of ATP-sensitive K+ channels during a bout of acute myocardial ischemia plays an important role in the emergence of ECG ST elevation. PMID- 8403331 TI - Entrainment of reentrant ventricular tachycardia in anisotropic rings of rabbit myocardium. Mechanisms of termination, changes in morphology, and acceleration. AB - BACKGROUND: Entrainment of ventricular tachycardia can either terminate or change the rate and/or morphology of ventricular tachycardia. The purpose of this study was to elucidate the underlying mechanisms by mapping of entrainment of ventricular tachycardia. METHODS AND RESULTS: In 10 Langendorff-perfused rings of anisotropic rabbit left ventricular epicardium created by a cryoprocedure, ventricular tachycardia with a cycle length of 167 +/- 17 milliseconds was induced by incremental pacing. During transient entrainment (10 stimuli), the circulating wave was extinguished by collision with the paced antidromic wave, whereas ventricular tachycardia was reset by the paced orthodromic wave. At shorter pacing intervals, the site of collision shifted deeper into the circuit. Entrainment at high rates (104 +/- 11 milliseconds) resulted in either termination (n = 54), a change in morphology (n = 8), or acceleration (n = 6) of ventricular tachycardia. Termination of ventricular tachycardia was due to complete (84%) or partial (16%) block of the paced orthodromic wave. Partial block induced microreentry within the circuit, resulting in a reflected echo wave that terminated ventricular tachycardia. A change in morphology of ventricular tachycardia was due to reversion of the direction of propagation of the circulating wave around the obstacle. Acceleration of ventricular tachycardia was caused by double-wave reentry induced by block of the paced antidromic wave. In 28 cases, the sequence of activation during entrainment was not stable but changed from beat to beat due to varying arcs of conduction block. Block occurred predominantly (86%) during slow transverse propagation. Before termination, local oscillations in interval occurred, resulting in a shortening of the last local interval at the site of block by 10 +/- 6 milliseconds. CONCLUSIONS: Termination of ventricular tachycardia by entrainment was due either to complete orthodromic block or to a reflected echo wave. A change in morphology occurred when the direction of the circulating wave reversed. Acceleration of ventricular tachycardia was due to initiation of double-wave reentry. All changes were preceded by conduction block during one or more stimuli at one or multiple sites in the circuit. Block occurred predominantly during slow transverse propagation and was preceded by local oscillations in interval at the site of block. PMID- 8403332 TI - Mechanism of spontaneous termination of stable atrial flutter in the canine sterile pericarditis model. AB - BACKGROUND: We tested the hypotheses that spontaneous termination of stable atrial flutter is directly related to spontaneous beat-to-beat cycle length oscillations and that block of the circulating reentrant wave front occurs in an area of slow conduction. METHODS AND RESULTS: We studied 30 episodes of spontaneous termination of stable atrial flutter induced by atrial stimulation in 11 conscious, nonsedated dogs with sterile pericarditis. Additionally, in 5 dogs, 14 episodes of spontaneous termination of stable atrial flutter were studied with a multisite mapping system to record simultaneously from 190 right atrial electrodes. In the conscious-state studies, atrial flutter cycle length oscillations began 6 +/- 1 (mean +/- SEM) beats before termination in 26 episodes, stable atrial flutter evolved into atrial fibrillation in 3 episodes, and no cycle length change occurred before termination in 1 episode. In the open chest studies, in all instances, spontaneous oscillations began 7 +/- 1 beats before termination. The only consistent oscillation pattern occurred for the last two beats: a long cycle length (149 +/- 9 milliseconds) followed by a much shorter cycle length (110 +/- 6 milliseconds) (P < .01). Activation maps demonstrated that all cycle length oscillations were explained by changes of conduction in an area(s) of slow conduction in the reentrant circuit. In two instances, the last (short) cycle length was associated with disappearance of an area of slow conduction. In all episodes, the last circulating reentrant wave front blocked in an area of slow conduction in the reentrant circuit. Although not tested, during the last beat, the very early arrival of the circulating reentrant wave front at an area of slow conduction suggests an important role for refractoriness, with head and tail interactions, resulting in block. CONCLUSIONS: Spontaneous termination of stable atrial flutter in the sterile pericarditis model (1) is preceded by beat-to-beat cycle length oscillations that result from changes in conduction in areas of slow conduction in the reentrant circuit and (2) results from block of the circulating reentrant wave front in an area of slow conduction. PMID- 8403333 TI - Effects of flecainide on ectopic atrial automaticity and conduction. AB - BACKGROUND: Previous studies have shown that class Ic antiarrhythmic agents are effective in suppressing ectopic atrial rhythms and accessory pathway conduction. METHODS AND RESULTS: To explore the potential mechanisms for their effectiveness, we investigated the concentration-dependent effects of the Ic agent flecainide acetate (0.5 to 10 micrograms/mL) on atrial ectopic automaticity and exit conduction in isolated rabbit tricuspid valves. This experimental model consists of three major cell types as defined anatomically and by intracellular recordings: pacemaker, transitional, and working atrial muscle. Simultaneous recordings from these cell types before and during flecainide superfusion (n = 7) showed that the drug produced a slight, concentration-dependent slowing of pacemaker-transitional conduction but elicited third-degree transitional-working atrial muscle block in six of seven preparations at 10 micrograms/mL. Flecainide caused a significant dose-dependent reduction in the initial phase of diastolic depolarization of pacemaker cells but produced only a small, biphasic change in spontaneous pacemaker cycle length. It also caused a significant prolongation in action potential duration in pacemaker and transitional cells and reduction in upstroke velocity in atrial cells. Of note in four additional preparations, flecainide caused a concentration-dependent upward shift in the strength-duration curve for atrial fibers. CONCLUSIONS: These data suggest that flecainide has little direct effect on ectopic atrial automaticity but rather causes exit conduction slowing and block between transitional and atrial muscle fibers. The mechanism for the induction of block is likely due to a decrease in atrial excitability creating a greater electrical load on generated impulses. PMID- 8403334 TI - Propranolol and lidocaine inhibit neural norepinephrine release in hearts with increased extracellular potassium and ischemia. AB - BACKGROUND: Propranolol and lidocaine are effective antiarrhythmic drugs in myocardial ischemia and infarction. As sympathetic nerve activation and norepinephrine release in ischemic hearts are arrhythmogenic, we tested the possibility that both agents inhibit neural norepinephrine release following sympathetic activation in the ischemic environment. METHODS AND RESULTS: The model used was an in situ perfused innervated rat heart. Norepinephrine release was induced by electrical stimulation of the left cervicothoracic stellate ganglion and analyzed using radioenzymatic assay or high-performance liquid chromatography. In normoxically perfused hearts, evoked norepinephrine release was not affected by either of the two agents at doses of 1 to 10 mumol/L when extracellular K+ concentration was 4 mmol/L but dose-dependently reduced at 10 mmol/L K+ (D,L-propranolol: -53 +/- 4% at 1 mumol/L and -64 +/- 6% at 10 mumol/L K+, lidocaine: -37 +/- 11% at 0.1 mumol/L, -67 +/- 5% at 1 mumol/L, and -75 +/- 6% at 10 mumol/L). At 10 mmol/L K+, norepinephrine release was not affected by timolol or atenolol (both 10 mumol/L but was equally inhibited by D- or L propranolol at 10 mumol/L (-56 +/- 5% and -53 +/- 9%, respectively), indicating a beta-blocking-independent mechanism. In hearts with metabolic acidosis (pH 6.85) at K+ of 4 mmol/L, neural norepinephrine release was also reduced by propranolol at 10 mumol/L (-37%). Finally, in hearts perfused with 4 mmol/L K+ and subjected to 6-minute periods of ischemia, neural norepinephrine release was similarly suppressed by D,L-propranolol (-38 +/- 6% at 0.1 mumol/L, -44 +/- 5% at 1 mumol/L, and -78 +/- 3% at 10 mumol/L) or lidocaine (-39 +/- 7% at 0.1 mumol/L, 58 +/- 9% at 1 mumol/L, and -91 +/- 3% at 10 mumol/L). CONCLUSIONS: These data indicate that propranolol and lidocaine inhibit neural norepinephrine release via a Na+ channel-blocking mechanism that is synergistic with changes induced by ischemia, primarily raised extracellular K+. This mechanism may contribute to the anti-ischemic and antiarrhythmic properties of both agents in acute myocardial ischemia, which induces increased extracellular K+ and sympathetic activation. PMID- 8403335 TI - Diminished contractile response to increased heart rate in intact human left ventricular hypertrophy. Systolic versus diastolic determinants. AB - BACKGROUND: Experimental studies indicate that in addition to diastolic dysfunction, hypertrophied myocardium can display depressed contractile responses, particularly at rapid heart rates, compounding reserve limitations. This study tests whether such abnormalities exist in intact human subjects at physiological paced rates and, if so, whether they are linked to simultaneous rate-dependent deterioration in diastolic function. METHODS AND RESULTS: Ten subjects with left ventricular hypertrophy (LVH) and 8 normal control subjects were studied. Most LVH patients presented with dyspnea and/or pulmonary edema and had concentric hypertrophy. Since rapid pacing simultaneously alters cardiac filling volumes and pressures, pressure-volume relation analysis was used to better define changes in contractile response. Patients were instrumented with a conductance catheter and micromanometer for pressure-volume data recording and a balloon occluder at the right atrial-inferior vena caval junction to vary filling and thus generate function relations. Data were obtained at baseline and at three atrial pacing rates (100, 120, 150 min-1). In addition, single-beat force interval data were used to indirectly examine calcium cycling kinetics. LVH subjects demonstrated baseline diastolic abnormalities, including prolonged relaxation, elevated end-diastolic pressure, and reduced chamber compliance. However, systolic function was similar to that in control subjects. With rapid pacing, normal subjects displayed a positive contractile response, whereas this was markedly diminished in LVH subjects. With abrupt termination of pacing and return to slower sinus rhythm, LVH subjects displayed greater initial potentiation followed by a more rapid decline than control subjects, suggesting abnormalities of calcium handling. Despite contractile abnormalities, diastolic function did not further deteriorate with rapid pacing and thus did not appear to be tightly linked to the systolic changes. CONCLUSIONS: Pacing stress in intact human LVH can result in systolic impairment superimposed on preexisting but not worsened diastolic dysfunction. Abnormal calcium handling probably contributes prominently to this response. PMID- 8403336 TI - Bystander cardiopulmonary resuscitation. Is ventilation necessary? AB - BACKGROUND: Prompt initiation of bystander cardiopulmonary resuscitation (CPR) improves survival. Basic life support with mouth-to-mouth ventilation and chest compressions is intimidating, difficult to remember, and difficult to perform. Chest compressions alone can be easily taught, easily remembered, easily performed, adequately taught by dispatcher-delivered telephone instruction, and more readily accepted by the public. The principal objective of this study was to evaluate the need for ventilation during CPR in a clinically relevant swine model of prehospital witnessed cardiac arrest. METHODS AND RESULTS: Thirty seconds after ventricular fibrillation, swine were randomly assigned to 12 minutes of chest compressions plus mechanical ventilation (group A), chest compressions only (group B), or no CPR (group C). Standard advanced cardiac life support was then provided. Animals successfully resuscitated were supported for 2 hours in an intensive care setting, and then observed for 24 hours. All 16 swine in groups A and B were successfully resuscitated and neurologically normal at 24 hours, whereas only 2 of 8 group C animals survived for 24 hours (P < .001, Fisher's exact test). One of the 2 group C survivors was comatose and unresponsive. CONCLUSIONS: In this swine model of witnessed prehospital cardiac arrest, the survival and neurological outcome data establish that prompt initiation of chest compressions alone appears to be as effective as chest compressions plus ventilation and that both techniques of bystander CPR markedly improve outcome compared with no bystander CPR. PMID- 8403337 TI - Augmented efficacy of external CPR by intermittent occlusion of the ascending aorta. AB - BACKGROUND: After prolonged cardiac arrest, conventional methods of closed-chest cardiac compression are ineffective. This is primarily because of failure to generate minimal threshold levels of coronary perfusion pressure for cardiac resuscitation. This report introduces a new option for cardiac resuscitation by use of a combination of intermittent ascending aortic balloon occlusion, aortic infusion, and precordial compression to increase the pressure gradient for coronary perfusion. METHODS AND RESULTS: Twenty anesthetized, mechanically ventilated, normovolemic domestic pigs were investigated. A 10F balloon catheter was advanced from the left femoral artery into the ascending aorta. Ventricular fibrillation was induced with an AC current delivered through an electrode catheter advanced into the right ventricle. Precordial compression was initiated after 7 minutes of untreated ventricular fibrillation. The animals were randomized to one of four groups: (1) balloon occlusion with proximal infusion of oxygenated saline, (2) balloon occlusion alone, (3) proximal aortic infusion together with epinephrine without balloon occlusion, and (4) injection of epinephrine without balloon occlusion or proximal infusion. For balloon occlusion, the balloon was inflated for 30 seconds during each minute of cardiopulmonary resuscitation. In the subsets of animals that received infusions, oxygenated saline (30 mL) was injected into the proximal aorta immediately after balloon occlusion. Epinephrine was used in two subsets: It was injected as a bolus in amounts of 30 micrograms/kg into the right atrium at 30 seconds after start of precordial compression and repeated as required to maintain coronary perfusion pressure within the range of 25 to 30 mm Hg. Defibrillation was attempted at 1 minute after start of precordial compression and at 1-minute intervals thereafter. Resuscitation attempts were continued until there was return of spontaneous circulation or for a total of 30 minutes after start of precordial compression. Coronary perfusion pressure generated by precordial compression was significantly increased after balloon occlusion. Each of 10 animals was successfully resuscitated and survived for 48 hours after balloon occlusion whether or not it was combined with infusion. Three of five animals were resuscitated by a combination of infusion and epinephrine in the absence of aortic occlusion, but none survived for 48 hours (P = .02). Only one epinephrine treated animal was successfully resuscitated and survived for 48 hours in the absence of balloon occlusion or infusion (P < .05). CONCLUSIONS: Ascending aortic balloon occlusion with or without proximal aortic infusion strikingly increased resuscitability and 48-hour survival after cardiac arrest under conditions when conventional methods failed. PMID- 8403338 TI - Hyperbaric oxygen limits infarct size in ischemic rabbit myocardium in vivo. AB - BACKGROUND: We explored the ability of increased oxygen pressure to modify necrosis in an open-chest rabbit model of myocardial ischemia and reperfusion. METHODS AND RESULTS: A branch of the left coronary artery was occluded for 30 minutes followed by 3 hours of reperfusion. Infarction was measured by triphenyl tetrazolium staining and expressed as a percentage of the ischemic zone. Untreated rabbits were ventilated with 100% oxygen at 1 atm absolute. Treatment animals were exposed to hyperbaric oxygen at 2.5 atm absolute. The 1.0-atm control hearts developed 41.5 +/- 4.6% infarction of the ischemic zone. Animals exposed to hyperbaric oxygen during ischemia only, reperfusion only, or ischemia and reperfusion had significantly smaller infarcts with respect to control animals (16.2 +/- 2.9%, 14.5 +/- 3.7%, and 9.8 +/- 2.7%, respectively; P < or = .01), indicating that they had been protected by the procedure. When hyperbaric oxygen was begun 30 minutes after the onset of reperfusion, no protection was seen (35.8 +/- 3.8%). CONCLUSIONS: We conclude that hyperbaric oxygen limits infarct size in the reperfused rabbit heart and that the effect can be achieved when hyperbaric oxygen is begun at reperfusion. PMID- 8403339 TI - The advent of adenovirus. Gene therapy for cardiovascular disease. PMID- 8403340 TI - Images in cardiovascular medicine. Epitome of cardiac tamponade. PMID- 8403341 TI - A 17-year-old woman with a history of sexually transmitted disease and productive cough who developed cardiogenic shock. PMID- 8403342 TI - Cardiovascular complications of non-insulin-dependent diabetes. The difficult search for causality. PMID- 8403343 TI - Serum cholesterol. Doing the right thing. PMID- 8403344 TI - Aspirin versus coumadin. PMID- 8403345 TI - Stability of plasma atrial natriuretic peptide. PMID- 8403346 TI - von Willebrand factor and atherosclerosis. PMID- 8403347 TI - T. Duckett Jones Memorial Lecture. Global assessment of rheumatic fever and rheumatic heart disease at the close of the century. Influences and dynamics of populations and pathogens: a failure to realize prevention? PMID- 8403348 TI - Socioeconomic factors and cardiovascular disease: a review of the literature. AB - Despite recent declines in mortality, cardiovascular diseases are the leading cause of death in the United States today. It appears that many of the major risk factors for coronary disease have been identified. Researchers are still learning about different modifiable factors that may influence cardiovascular diseases. Socioeconomic status may provide a new focus. The principal measures of SES have been education, occupation, and income or combinations of these. Education has been the most frequent measure because it does not usually change (as occupation or income might) after young adulthood, information about education can be obtained easily, and it is unlikely that poor health in adulthood influences level of education. However, other measures of SES have merit, and the most informative strategy would incorporate multiple indicators of SES. A variety of psychosocial measures--for example, certain aspects of occupational status--may be important mediators of SES and disease. The hypothesis that high job strain may adversely affect health status has a rational basis and is supported by evidence from a limited number of studies. There is a considerable body of evidence for a relation between socioeconomic factors and all-cause mortality. These findings have been replicated repeatedly for 80 years across measures of socioeconomic level and in geographically diverse populations. During 40 years of study there has been a consistent inverse relation between cardiovascular disease, primarily coronary heart disease, and many of the indicators of SES. Evidence for this relation has been derived from prevalence, prospective, and retrospective cohort studies. Of particular importance to the hypothesis that SES is a risk factor for cardiovascular disease was the finding by several investigators that the patterns of association of SES with coronary disease had changed in men during the past 30 to 40 years and that SES has been associated with the decline of coronary mortality since the mid-1960s. However, the declines in coronary mortality of the last few decades have not affected all segments of society equally. There is some evidence that areas with the poorest socioenvironmental conditions experience later onset in the decline in cardiovascular mortality. A number of studies suggest that poor living conditions in childhood and adolescence contribute to increased risk of arteriosclerosis. Some of these studies have been criticized because of their nature, and others for inadequate control of confounding factors.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8403349 TI - Cardiovascular disease in women. PMID- 8403350 TI - Abstracts from the 66th Scientific Sessions of the American Heart Association. Atlanta, Georgia, November 8-11, 1993. PMID- 8403351 TI - Transesophageal echocardiography: usefulness increasing. PMID- 8403353 TI - The diagnostic challenge of Behcet's disease. PMID- 8403352 TI - Menopause: managing the associated risks. PMID- 8403354 TI - The role of blood viscosity in the development and progression of coronary artery disease. AB - BACKGROUND: Although a great deal of attention has been directed to the risk factors for atherosclerotic coronary artery disease, relatively little research has been focused on the role of the circulating blood itself. OBJECTIVES: To review the possible role of increased plasma viscosity and decreased cellular deformability in the pathogenesis of atherosclerotic coronary artery disease. SUMMARY: Increased plasma viscosity or decreased erythrocyte deformability may reduce blood flow in areas where blood flow is already low, such as at arterial branch points and bifurcations as well as in advanced coronary artery narrowing. Further, these factors may favor the development and progression of atherosclerotic coronary artery disease through direct mechanical effects on either the vascular endothelium or existing atheromatous plaques. Beyond their potential direct effects, each may represent a common link between recognized risk factors and the resulting disease process. CONCLUSIONS: Accumulating evidence suggests that increased blood viscosity is an independent risk factor for atherosclerotic heart disease and its complications. PMID- 8403355 TI - The role of azithromycin and clarithromycin in clinical practice. AB - BACKGROUND: Azithromycin and clarithromycin are the newest approved macrolide antibiotics. OBJECTIVE: To review the pharmacology, microbiology, and clinical utility of these agents. SUMMARY: These agents have distinct advantages over erythromycin, including an improved pharmacokinetic profile, less toxicity, and a wider spectrum of activity. They are approved for the treatment of respiratory tract infections and uncomplicated skin and skin-structure infections associated with specific organisms. Azithromycin is also indicated for the treatment of nongonococcal urethritis. In addition, these agents may be useful in the treatment of toxoplasmosis, mycobacterial disease, Lyme disease, and legionellosis. Clarithromycin and azithromycin have lower rates of gastrointestinal side effects than erythromycin. CONCLUSIONS: Although clarithromycin and azithromycin show promise in the treatment of some less common infections, they should be considered alternatives to conventional agents in the treatment of respiratory tract, skin, and skin-structure infections caused by the usual pathogens. The expense of these agents may be prohibitive for routine use. PMID- 8403356 TI - Pheochromocytoma: current diagnosis and management. AB - BACKGROUND: Pheochromocytoma is a catecholamine-secreting tumor of chromaffin cells that causes hypertension. OBJECTIVE: To review the clinical presentation, diagnosis, and treatment of this disease. SUMMARY: Pheochromocytoma can mimic a number of other diseases, making recognition difficult. Hypertension may be paroxysmal or sustained. The signs and symptoms of pheochromocytoma are mostly due to hypercatecholaminemia, hypertension, complications, or coexisting diseases; however, measurements of catecholamines and their metabolites in the plasma and urine may be normal between "attacks", and other conditions can elevate these values. The clonidine suppression test confers specificity to the clinical and laboratory findings, and magnetic resonance imaging is the most reliable method of locating a tumor. Surgical resection is successful in 90% of patients; however, the disease is fatal if it is not detected and treated. CONCLUSIONS: Pheochromocytoma should be suspected in patients with paroxysmal or sustained hypertension, particularly if symptoms are present. PMID- 8403357 TI - Primary aldosteronism: new approaches to diagnosis and management. AB - Primary aldosteronism remains a diagnostic challenge. Certain immunoassay techniques, simplified diagnostic testing, and the introduction of sensitive imaging techniques have facilitated the diagnosis, but obstacles that remain include a lack of optimal screening methods, low sensitivity and specificity of current diagnostic tests, and a growing number of etiological subgroups. A rational approach to the diagnosis of primary aldosteronism is described, as is the differentiation of the surgically correctable lesion (adenoma) from the other etiological subgroups. PMID- 8403358 TI - Coronary artery disease in women: a risk-factor analysis. AB - BACKGROUND: Risk factor modification is important in preventing coronary artery disease; however, risk factors for coronary artery disease have been studied mostly in men, not women. OBJECTIVE: To examine risk factors for coronary artery disease in women. METHODS: We retrospectively reviewed the records of all adult women who underwent their first-ever cardiac catheterization at our institution in 1983. Risk factors in women with angiographic evidence of coronary atherosclerosis were compared with risk factors in women without angiographic evidence of coronary artery disease. RESULTS: Risk factors identified included age, diabetes mellitus, hypertension for more than 5 years, hyperlipidemia, smoking, and familial coronary artery disease. CONCLUSIONS: Risk factors for coronary artery disease in women are similar to those of men. PMID- 8403359 TI - Coronary artery disease in young women: risk factor analysis and long-term follow up. AB - BACKGROUND: Heart disease in the leading cause of death in women of all ages in the United States, but data on coronary disease in young women remains sparse. OBJECTIVE: To identify and follow up a cohort of young women referred for the evaluation of coronary disease. METHODS: Retrospective review of the medical records. RESULTS: Thirty-two women younger than 31 years met the entry criteria. The average age was 28 +/- 2.4 years, 28% had insulin-dependent diabetes mellitus, 38% had hypertension, 6% had congenital heart disease, 38% had a family history of coronary artery disease, 72% were smokers, and 28% used oral contraceptives. Serum cholesterol levels were > 5.17 mmol/L (200 mg/dL) in 71%, and the mean cholesterol level was 6.70 +/- 2 mmol/L (259 +/- 78 mg/dL). Resting electrocardiographic results were abnormal in 28 women (88%), 22 of whom had evidence of transmural myocardial infarction. Follow-up averaged 9.8 +/- 6.4 years. Five patients died, all of whom had hypertension, and 4 of whom had diabetic nephropathy and required dialysis. CONCLUSIONS: Risk factors for coronary disease in young women include hypertension, hypercholesterolemia, diabetes mellitus, familial coronary disease, and smoking. Long-term prognosis is excellent for those without advanced diabetes mellitus and renal failure. PMID- 8403360 TI - Stereotactic and computer-assisted neurosurgery at the Cleveland Clinic: review of 501 consecutive cases. AB - BACKGROUND: Stereotactic and computer-assisted techniques have revolutionized the diagnosis and treatment of many disorders of the brain by directing surgical instruments, the surgeon's hands, or focused radiation to an imaged target along predefined routes. METHODS: This report reviews the basic principles and techniques of stereotactic and computer-assisted procedures in 501 consecutive procedures. RESULTS: Procedures performed include biopsy; decompression of cysts, hematomas, ventricles, and abscesses; "point," computer-assisted volumetric, and frameless stereotactic resection of brain lesions; placement of depth electrodes; treatment of movement or pain disorders; implantation of radioactive seeds in malignant tumors; and radiosurgery. CONCLUSIONS: Overall, image-directed stereotactic neurosurgery proved safe, accurate, and versatile. PMID- 8403361 TI - The Mona Lisa smiles: impact of risk factors for coronary artery disease in women. PMID- 8403362 TI - Self-test. Early-onset emphysema. PMID- 8403363 TI - F. Merlin Bumpus, PhD. PMID- 8403364 TI - Endothelin concentrations in respiration-related structures of the medulla during the perinatal period of the rat. AB - Endothelin-like immunoreactivity (ET-LI) was quantified in the developing (foetal and postnatal) rat brain stem and cerebellum using radioimmunoassay. The brain stem structures chosen for this study were (a) dorsal medullary region (DMR) including the region of nucleus tractus solitarius where the peripheral chemoreceptor afferents are known to terminate, (b) ventral medullary region (VMR) where the central chemoreceptors are thought to be located and (c) cerebellum (CER), as a control area. Compared to the prenatal period, significantly elevated concentrations of ET-LI were detected in the early postnatal period and thereafter the concentrations decreased: DMR and VMR: in comparison to the prenatal concentrations, a two-fold increase was found on the day of birth which further increased significantly (P < 0.001) on postnatal day 1 only in the region of DMR; CER: low concentrations of ET-LI were found in the early postnatal period which were not significantly different from the prenatal values. No ET-LI could be detected in any of the three regions in the adult rats. The results are discussed in view of the hypothesis that (1) endothelin appears to play an important role in the perinatal period and (2) it is involved in the chemoreceptor pathway. PMID- 8403365 TI - Origin and migration of trochlear, oculomotor and abducent motor neurons in Petromyzon marinus L. AB - The development of the ocular motor system was studied in 3- to 6-year old larval lampreys with two different retrograde tracers. Motor neurons of the oculomotor and trochlear nuclei are situated closely to one another in younger larvae. Cases in which only trochlear neurons were labelled revealed trochlear motor neurons scattered from the midbrain tegmentum through the anterior medullary velum. We believe this distribution reflects the place of final mitosis (midbrain tegmentum) and subsequent migration (anterior medullary velum) of lamprey trochlear motor neurons. Evidence is also presented for contralateral migration of oculomotor motor neurons and for ventrolateral migration of abducent motor neurons. The distances covered by migrating ocular motor neurons range from 100 to 150 microns in small larvae; these are distances that could be covered easily during the several years duration of larval development in lampreys. PMID- 8403366 TI - A monoclonal antibody, raised against rat embryo hippocampus, identifies a nuclear protein enriched in hippocampal and other neurons of the adult rat brain. AB - We report characteristics of a monoclonal antibody (MAb) produced by immunizing mice against rat embryo hippocampus, and its cellular distribution in the brains of adult and embryonic rats. This antibody, designated as MAb 4A4, brightly stained granular and pyramidal neurons of the adult rat hippocampus, as well as some cortical neurons. Also, MAb 4A4 labeled granular and Purkinje cells of the cerebellum with less intensity. While glial cells were labeled relatively faintly. At embryonic day 18 in the rat, most brain neurons, primitive neuroepithelium, connective tissues and glia were labeled with this antibody, indicating that the expression of 4A4 antigen is regulated developmentally. The 4A4 antigen appeared to be localized to the nucleus of cells except in choroid plexus in which the focal membrane staining was observed. The nuclear localization of 4A4 antigen was further confirmed by the staining of cultured cell lines with MAb 4A4. Western blot analysis demonstrated a single band of the 4A4 antigen from cultured cells, with an apparent molecular weight of 220 kDa. Both the molecular weight and the distribution of the 4A4 antigen in the embryonic and adult rat brain and cultured cells suggest that this antigen is a novel nonhistone nuclear type, which is preferentially expressed in neurons of the rodent brains and is under developmental regulation. PMID- 8403367 TI - Postnatal development of cholinergic presynaptic inhibition in rat hippocampus. AB - Muscarinic depression of field potential excitatory postsynaptic potentials (fEPSPs) in striatum radiatum of area CA1 was compared in hippocampal slices from rats of different ages. Bath application of 4 microM muscarine reversibly depressed the fEPSP slope by 68.4% in slices from adult animals (P43-P60), but caused only a 32.2% depression in slices from P5-P7 animals. The magnitude of the depression increased with age during the first postnatal month. Reduced sensitivity of excitatory synaptic transmission to cholinergic depression during postnatal development could be one factor contributing to the hyperexcitability of immature hippocampus. PMID- 8403368 TI - Odor-induced sexual maturation and expression of c-fos in the olfactory system of juvenile female mice. AB - Exposure to the urine or soiled bedding odors of novel adult males is known to accelerate puberty in juvenile female mice. To determine what part of the olfactory system is activated by these odors, the expression of c-fos in the main olfactory bulb (MOB) and the accessory olfactory bulb (AOB) of juvenile female mice was examined after their exposure to male-soiled bedding, peppermint odor or their own bedding. Fos-like immunoreactivity was found throughout the AOB of the juvenile female mice exposed to the bedding odors of adult males for 3 h. In contrast, dense staining was found in the granular cell layer of the MOB of mice exposed to peppermint odors for 3 h, whereas mice exposed to their own bedding failed to show immunostaining in the AOB and only slight or no staining in the MOB. These results indicate that social odors stimulate the expression of c-fos in the AOB while non-social odors activate the MOB. This method allows the identification of individual cells activated by the different odors and will be useful in locating other areas of the brain involved in the neuroendocrine changes underlying odor-induced precocious puberty. PMID- 8403369 TI - Ontogeny of the sexually dimorphic area of the gerbil hypothalamus. AB - The sexually dimorphic area (SDA) of the gerbil hypothalamus is a set of cell groups in the medial preoptic area that is essential for masculine sexual behavior and implicated in the hormonal control of scent making and ultrasound production. The adult SDA shrinks after gonadectomy unless the gerbils receive testosterone. So does the SDA pars compacta, a small cell group in the SDA of males that is seldom seen in females. Here, development of the SDA and SDApc, and of a second, small, compact cell group, the cmSDApc, that lies caudal and medial to the SDApc, is described. Development of the SDApc and cmSDApc was studied quantitatively by assessing their incidence and volume in both sexes from birth (PND 1) to adulthood (PND 150). The volume of the entire SDA was studied from PND 45 to 150. In male gerbils, puberty begins around PND 40 and is complete by PND 90-120. The male SDA enlarged relative to the cross-sectional area of the hypothalamus as puberty began, but the female SDA did not. The SDApc was present in virtually all gerbils at birth and was the same size in both sexes. Over the next two weeks, the SDApcs of females disappeared while those of males persisted and doubled in size. Like the SDApc, the cmSDApc was larger and more common in males than in females, but it became smaller and less prevalent in both sexes during the first two weeks after birth. PMID- 8403370 TI - The beta-endorphin response to prenatal stress during postnatal development in the rat. AB - We examined the hypothalamic beta-endorphin (B-END) levels in offspring (postnatal day 10) from stressed female rats in different period of gestation: group 1, restraint stress from day 2 to 6; group 2, from day 7 to 11 and group 3, from day 2 to 16. The hypothalamic levels of B-END in the offspring from group 1 were significantly (P < 0.001) higher to that obtained in the control. Similar results were obtained in the group 2. However, in rat pups from female stressed during 15 days of gestation hypothalamic levels of B-END were similar to control group. PMID- 8403371 TI - Synapse formation in dissociated cell cultures of embryonic chick cerebral neurons. AB - The development of synapses was confirmed in the primary cultures of dissociated cerebral cortex neurons from chick embryos. Whole-cell patch clamp recording applied to dissociated neurons from 6- to 12-day-old embryos revealed that these neurons form functional synapses. In these cultures, both excitatory and inhibitory postsynaptic responses were observed. Synaptogenesis in our cultures seemed to be in proportion to the embryonic equivalent days, which are the sum of the days in incubation and culture. PMID- 8403372 TI - Modulation of neuropeptide Y expression in rat brain neuronal cultures. AB - Neuropeptide Y (NPY) and its encoding mRNA were measured in neurons co-cultured from rat basal forebrain and cerebral cortex. NPY was synthesized and released in a manner consistent with secretion-synthesis coupling; depolarization increased each in a calcium-dependent manner. The accumulation of NPY encoding mRNA was elevated by a muscarinic receptor blocker, without changes in transmitter release or peptide synthesis, thereby consistent with a membrane potential-independent mechanism. Changes in intrinsic muscarinic transmission could nonetheless be expressed rapidly as an elevation in NPY levels by depolarizing the neurons subsequent to muscarinic receptor blockade. This depolarization-induced elevation of NPY subsequent to muscarinic receptor blockade was dependent on the presence of extracellular calcium ions. Forskolin and pertussis toxin also increased NPY encoding mRNA levels in a manner that was not additive with muscarinic receptor blockade. These results suggest that one or more muscarinic receptors may tonically modulate NPY synthesis via changes in adenylate cyclase activity, providing a model for the non-homeostatic modulation of neuropeptide turnover. PMID- 8403373 TI - Intrastriatal cerebellar grafts: differentiation of cerebellar anlage and sprouting of Purkinje cell axons. AB - Pieces of cerebellar primordia were obtained from G16 (day 16 of gestation) rat fetuses and stereotaxically injected into the striatum of adult Wistar rats. The transplants were allowed to integrate with the host brain for 2 h up to 6 months after implantation. Ninety four out of 105 transplants perfectly integrated with the host brain (90%) and established the typical trilaminar histoarchitecture of cerebellar cortex. The transplants were sufficiently vascularized. Vessels seen within the grafts provided all ultrastructural elements of a blood-brain barrier. Light microscopic evaluation of graft development showed no considerable retardation of cerebellar histogenesis. Electron microscopic examination disclosed normal ultrastructure of cerebellar neurons, as well as elements of regular synaptic organization. The topic of efferent graft-to-host projections was investigated 2.5 months after transplantation using the monoclonal Purkinje cell marker anti-Leu-4 (CD3). This method allowed us to detect immunoreactive, morphologically intact axons of grafted Purkinje cells running over long distances (at least 500 microns) within the host striatum. Whilst afferent but in no case efferent connections of heterotopic cerebellar transplants had been demonstrated elsewhere, we could now prove the reciprocal modus of graft-host interaction with heterotopic cerebellar grafts. PMID- 8403374 TI - Effect of astroglial degeneration on the blood-brain barrier to protein in neonatal rats. AB - Recent in vitro studies have suggested that astrocytes may be responsible for the induction of several blood-brain barrier (BBB) characteristics. To examine this hypothesis in an in vivo situation, we have investigated the effect of chronic astrocytic deprivation on the BBB to proteins in neonatal rats. Intraperitoneal injections of the gliotoxin 6-aminonicotinamide (6-AN) resulted in cytotoxic edema with subsequent necrobiosis of differentiated astrocytes and oligodendrocytes throughout the CNS. Animals were sacrificed 1-5 days after chronic exposure to 6-AN during the first postnatal week. Animals sacrificed 24 h after the final injection of 6-AN had the greatest depletion of perivascular astroglia. The BBB to exogenous protein, examined by intravascular administration of horseradish peroxidase, remained intact, as did the BBB to endogenous protein as determined by immunocytochemical detection of rat serum albumin. In no case was any leakage of protein found other than in areas that do not normally possess BBB characteristics. These data show that CNS endothelial cells retain BBB characteristics without a full complement of astrocytic contacts. Since the astroglial cytoplasm was destroyed and only membrane fragments remained, we suggest that factors continuously produced by astroglia cannot be responsible for the induction and maintenance of the BBB to protein, but that substances produced during the prenatal period may be the primary determinant of endothelial phenotype. PMID- 8403375 TI - Recovery of [3H]acetylcholine synthesis after AF64A-treatment in primary, neuron enriched, rat septal-hippocampal and striatal cultures. AB - Neuron-enriched cultures prepared from several different rat brain regions were incubated with 10 microM or 30 microM monoethylcholine mustard aziridinium ion (AF64A) under conditions (1 h, 37 degrees C in Krebs Ringer buffer) that reduced acetylcholine (ACh) synthesis coupled to high-affinity choline uptake, without affecting choline acetyltransferase activity. Co-cultures of septum-hippocampus and cultures of striatum were similarly sensitive to the AF64A-induced inhibition of ACh synthesis. However, ACh-synthesis recovered more rapidly in the striatal cultures than in septal-hippocampal co-cultures after AF64A washout. In septal hippocampal co-cultures, neither tunicamycin (20 micrograms/ml) nor cycloheximide (0.5 microgram/ml) had any effect on the basal synthesis of ACh or its recovery within 24 h following 10 microM AF64A washout. However, the recovery of ACh synthesis in these co-cultures after 30 microM AF64A-washout was blocked by either tunicamycin or cyclohexamide. Neither tunicamycin nor cyclohexamide interfered with ACh-synthesis recovery after washout of 30 microM AF64A in striatal cultures. These studies suggest that the turnover of high-affinity choline transporters can be modulated in a brain-region specific manner in intact primary neuronal cultures. PMID- 8403376 TI - Phrenic and recurrent laryngeal motoneuron activities during hyperoxia and hypoxia in piglets. AB - We hypothesized that synchronization of inspiratory motoneurons may involve inputs from two central pattern generators (CPG): one characterized by medium frequency (< 50 Hz) and the other by high-frequency oscillations (> or = 50 Hz). We studied phrenic and recurrent laryngeal nerve activities recorded during hyperoxia and hypoxia in Saffan anesthetized, paralyzed, and artificially ventilated piglets. Spectral analyses, derived from the full as well as partitioned halves of inspiration, showed that phrenic and recurrent laryngeal discharges contained peaks in the medium-frequency band, which were indicative of common inputs. The phrenic spectra of many animals had peaks in the high frequency band; such peaks were uncommon in recurrent laryngeal spectra; consequently, correlated activities corresponding to high-frequency oscillations were not usually observed. Thus, it is likely that acquisition of modulating inputs from a high-frequency CPG may emerge in an age-dependent manner in different motoneuron pools. During hypoxia, both phrenic and recurrent laryngeal discharges were facilitated as shown by increases in both the amplitudes of signal-averaged histograms and the magnitudes of their respective power spectral activities. Also, there was a significant increase in the values of phrenic recurrent laryngeal coherence estimates in the medium-frequency region. Hence, medium-frequency oscillations are more apparent in early development, perhaps to facilitate synchronization of inspiratory motoneuron activities, especially under conditions of increased chemical drive. PMID- 8403377 TI - Neurochemical alterations in Rett syndrome. AB - The Rett syndrome (RS) is a neurological disorder associated with severe mental deficiency and neurological manifestations of cortical and extrapyramidal dysfunction. The present report is (1) a postmortem brain study that compares the levels of choline acetyltransferase (ChAT) activity and the binding density of selected neurotransmitter receptors in four cases of RS and five normal controls of similar age and (2) a study of cerebrospinal fluid (CSF) concentrations of the endogenous tridecapeptide neurotensin in 12 RS patients and 8 controls of similar age. The level of ChAT activity was lower in many cortical and subcortical regions in the RS brains as compared to control levels. The number of NMDA, AMPA, mu opioid and neurotensin binding sites, as well as CSF concentrations of neurotensin, did not differ significantly from control levels. The results suggest that changes in specific neurotransmitter systems, particularly cholinergic neurons, in the thalamus, cerebellum and basal ganglia may underlie the progressive deterioration in motor and cognitive function characteristic of this disorder. PMID- 8403378 TI - Neurite promoting activity of insulin, insulin-like growth factor I and nerve growth factor on spinal motoneurons is astrocyte dependent. AB - Mouse motoneurons were isolated from dissociated E15 mouse spinal cord and grown on polyornithine-coated round coverslips in a growth medium (DMEM/F12) supplemented with progesterone, trans-ferrin, selenium, horse serum and muscle extract. Astrocytes from newborn mouse neopallium were grown on rectangular coverslips. The motoneuron neurite growth was determined at day 8 of culture by counting, using the light microscope, the intersections produced by neurites radiating from the perikaryon placed centrally in a graticule eyepiece of concentric circles. The mean intersections for cultures without addition of astrocytes, insulin, insulin-like growth factor I (IGF-I) or nerve growth factor (NGF) was 12.6 +/- 0.8. When astrocytes on a separate coverslip were introduced from day 1, there was a small increase in neurite growth (16.3 +/- 0.9). The neurite growth was further increased significantly with the addition of insulin (27.3 +/- 1.4), IGF-I (31.5 +/- 1.4) or NGF (21.8 +/- 1.1) to cultures with astrocytes. Insulin, IGF-I or NGF in the absence of astrocytes did not greatly increase the neurite growth. We conclude that insulin, IGF-I and NGF promote neurite growth through some interactions with astrocytes. PMID- 8403379 TI - Early axon outgrowth of retinal ganglion cells in the fetal rhesus macaque. AB - Employing retinal explants and retrograde transport techniques, we studied the formation of the arcuate fascicles by examining the growth of the central retina, the emergence of the adult fiber layer pattern, and the projections of retinal ganglion cells in the central and peripheral retina. Sixty days prior to foveal pit formation, the distance from the incipient fovea to the optic disk was equal to the adult, even though the retinal area was only 8% of the adult. Arcuate fibers, at this age, were observed to avoid the incipient fovea, with no fascicles and few axons projecting over this region. A small population of 15.2% of the ganglion cells located within 2 mm of the incipient fovea possessed an axon with an aberrant trajectory that wound around and projected 50 to several hundred microns away from the optic disk, compared to only 3% at other retinal locations. The incidence of disorder decreased with increasing fetal age, establishing mature values in late fetal periods. These findings suggest that the area of the central retina does not increase after embryonic day 60 and that guidance factors are present that allow outgrowing ganglion cell axons to distinguish and avoid that portion of the retina that will become the fovea. PMID- 8403380 TI - Postnatal development of central nervous alpha 2-adrenergic binding sites: an in vitro autoradiography study in the tree shrew. AB - The postnatal development of the alpha 2-adrenoceptor pattern was investigated by in vitro receptor autoradiography with the antagonist [3H]rauwolscine in the brains of tree shrews (Tupaia belangeri). At birth, high numbers of [3H]rauwolscine binding sites are diffusely distributed in the whole brain with exception of the neocortex which is very weakly labeled at this time. While the number of [3H]rauwolscine binding sites in the cerebellum decreases to low levels during the first three postnatal weeks, several brain regions show a significant increase in binding sites which are progressively concentrated in distinct nuclei. In the medulla oblongata, the diffuse labeling pattern changes so that binding sites become centralized in the dorsomedial nuclei. In the pons, similar changes can be observed with a moderate labeling of the locus coeruleus on postnatal day 10 and a strong labeling in the adult. In the thalamus, a transient appearance of high numbers of [3H]rauwolscine binding sites can be observed during the second and third postnatal week in specific nuclei. In the preoptic area and hypothalamus, there are only minor postnatal changes but the numbers of [3H]rauwolscine binding sites decrease between postnatal day 5 and adulthood. The high number of binding sites in the limbic system does not significantly change after birth. In the neocortex and the superior colliculus, the [3H]rauwolscine labeling pattern which is characteristic for the adult is achieved not before the third postnatal week. Competition experiments demonstrate that [3H]RAUW binds with high affinity to alpha 2-adrenoceptors in the postnatal as well as in the adult brain. Therefore, this study demonstrates region specific developmental profiles of the pattern of alpha 2-adrenoceptors in the postnatal tree shrew brain. PMID- 8403381 TI - Transient expression of calbindin-D28k immunoreactivity in layer V pyramidal neurons during postnatal development of kitten cortical areas. AB - Calbindin-D28k is a 28 kDa calcium binding protein that has been shown to colocalize with a specific subpopulation of gamma-aminobutyric acid inhibitory interneurons in mammalian neocortex. We have examined the ontogeny of calbindin in neonatal kitten cortex in areas 17,18,19,7, medial and lateral suprasylvian visual areas, splenial visual area and cingulate cortex from the day of birth (P0) through maturation of the brain (P101). Transient staining of immature layer V pyramidal cells was seen in kittens six weeks old and younger. This transient staining of pyramidal cells was most intense and the stained neurons were most numerous in cingulate cortex. Apical dendrites of pyramidal cells in cingulate cortex were prominently stained and could be followed to layer I, where they were seen to branch extensively. There were very few calbindin immunoreactive pyramidal cells in primary cortical areas postnatally. Transient staining in extrastriate visual cortical areas disappeared first from the lateral suprasylvian areas, and persisted longest in area 7. Pyramidal neurons in the cingulate gyrus expressed calbindin longest, but calbindin expression by pyramidal neurons ceased by the sixth postnatal week in all areas of the brain. PMID- 8403382 TI - Transcriptional activation of ornithine decarboxylase in adult and neonatal hippocampal slices. AB - Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine synthesis and is regulated by both transcription-dependent and transcription-independent mechanisms. We compared the effects of asparagine, an amino acid previously shown to increase ODC activity in adult hippocampal slices, on ODC mRNA and activity in adult and neonatal hippocampal slices. In addition, we evaluated the effects of asparagine on ODC activity following seizure activity elicited by systemic administration of kainic acid (KA) in both adult and neonatal rats. Asparagine produced an increase in ODC gene expression and activity in both adult and neonatal hippocampal slices. The increase in ODC activity elicited by asparagine in hippocampal slices was the same in control animals as in animals sacrificed 16 h after KA-induced seizure activity. The asparagine-elicited increase in ODC activity in neonatal and adult hippocampal slices was blocked by the RNA synthesis inhibitor, actinomycin D. Finally, polyamines produced an inhibition of ODC activity in neonatal hippocampal slices. The results indicate that the regulation of the expression and activity of ODC is similar in neonatal and adult hippocampus. PMID- 8403383 TI - Neural aromatase activity in a marsupial, the gray short-tailed opossum (Monodelphis domestica): ontogeny during postnatal development and androgen regulation in adulthood. AB - Neural aromatase activity (AA) was measured in gray short-tailed opossums (Monodelphis domestica) on the day of birth and at selected ages through adulthood. In adulthood, regulation by testicular androgens was examined. In mixed-sex samples of whole brain, AA was present at birth and increased until postnatal day (PD) 16. In hypothalamus-preoptic area (HPOA), significantly higher levels of AA were seen in animals on PD 16 than on PD 30 and PD 30 animals had higher levels of AA than all older ages including adults. Significant sex differences in HPOA AA with male levels higher, were seen only on PD 16 and in adulthood. While lower overall than in HPOA, AA was present also in cerebral cortex (CX). In CX, AA was higher on PDs 16 and 30 than at older ages. Significant sex differences in CX AA were observed only in adulthood. One week following castration in adulthood, AA dropped significantly in CX but not in HPOA. These findings are compared with those obtained from other marsupial and eutherian mammals with reference to the possible significance of AA in sexual differentiation of the gray opossum brain. PMID- 8403384 TI - Localization and ontogeny of growth hormone receptor gene expression in the central nervous system. AB - There is literature evidence that both growth hormone (GH) and its mediator, insulin-like growth factor 1 (IGF-1), are able to act upon neuronal and glial cells in the brain. We report here the location of the GH receptor in the brain of the rat and rabbit. Receptor distribution was determined by immunohistochemistry with GH receptor/binding protein (BP) specific monoclonal antibodies and by in situ hybridization with a [35S]riboprobe. GH receptor/BP immunoreactivity in the rat was most prominent in the neonate and declined with postnatal age. Receptor immunoreactivity was generalised with variation in immunoreactivity in regional areas. In the rat, strongest immunoreactivity was seen in layers 2, 3, 5 and especially layer 6 of the cerebral cortex, in neurones of the thalamus and hypothalamus, in Purkinje cells of the cerebellum, in neurones of the trapezoid body of the brainstem, and in retinal ganglion cells. Glial cells, notably astrocytes were also strongly reactive, along with ependyma of the choroid plexus, ventricular lining and pia mater. In the neonatal rabbit, strongest immunoreactivity was evident in layers 2 and 3 of the cerebral cortex, in pyramidal cells of the hippocampus, and in neurones of the inferior and superior colliculi, brain stem reticular formation, dorsal thalamus and hypothalamus. A similar distribution of GH receptor mRNA was seen by in situ hybridization. The ontogeny of GH receptor/BP mRNA in whole rat brain was quantified by solution hybridization-RNAse protection assay. Contrary to its ontogeny in the liver (Endocrinology, 113 (1983) 1325-1329) receptor mRNA decreased with postnatal age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403385 TI - Synaptophysin and synaptoporin expression in the developing rat olfactory system. AB - The expressions of two closely related synaptic vesicle antigens synaptophysin and synaptoporin were examined in the olfactory system of the adult rat and during pre- and postnatal development. In the adult, immunocytochemistry showed that the continuously regenerating olfactory receptor neurons (primary neurons) produce both synaptophysin and synaptoporin which were localized in the cell bodies of the receptor neurons in the olfactory epithelium, their dendrites, axonal processes in the olfactory nerve and their terminals in the olfactory bulb glomeruli. Furthermore, ultrastructural analysis revealed synaptophysin- and synaptoporin-immunoreactivities associated with synaptic vesicles in most olfactory receptor axonal terminals impinging on dendrites of the mitral and tufted neurons (secondary neurons in the olfactory bulb circuitry) in the olfactory glomeruli. In like manner, tufted neurons, granule and periglomerular neurons (interneurons in the olfactory bulb circuitry) express both synaptophysin and synaptoporin. In contrast, mitral neurons expressed only the synaptophysin antigen which was likewise associated with mitral axonal terminals in their target the olfactory cortex. The patterns of synaptophysin and synaptoporin expressions in mitral neurons (synaptophysin only) and tufted neurons (synaptophysin and synaptoporin) were similar in prenatal, postnatal and adult rats as revealed by immunocytochemistry and in situ hybridization. However, the biosynthesis of synaptophysin and synaptoporin by granule and periglomerular neurons, olfactory bulb interneurons, occurred mainly postnatally. PMID- 8403386 TI - Allocation of mouse cerebellar granule cells derived from embryonic ventricular progenitors--a study using a recombinant retrovirus. AB - Granule cells of the mammalian cerebellar cortex originate from embryonic progenitors present in the ventricular germinal layer. To investigate the allocation fate of these ventricular progenitors in the mouse, we labeled a few of them on embryonic day 13 with a recombinant retrovirus carrying lacZ which encodes E. coli beta-galactosidase (beta-gal), and the labeled cells in the postnatal cerebellar cortex were detected by beta-gal histochemistry. In the postnatal cerebellar cortex, the virally-labeled beta-gal+ granule cells formed discrete clusters. These clusters were not compactly packed with the beta-gal+ cells, and there was intermingling with beta-gal- granule cells. Neither beta gal+ Purkinje cells nor glia were found to be included in the clusters. Most of the granule cell clusters were incompatible with the functional areas of the cortex. These results suggest: (1) granule cells derived from individual ventricular progenitors are allocated in clusters and are not extensively dispersed, (2) granule cells descended from one progenitor may mix with their neighbors that are descended from another progenitor, (3) the allocation fate of the ventricular progenitors of granule cells is not restricted to the functional areas of the cerebellar cortex. PMID- 8403387 TI - Regional differences in the ontogenetic pattern of kynurenine aminotransferase in the rat brain. AB - The ontogenetic pattern of kynurenine aminotransferase (KAT), the biosynthetic enzyme of the neuroprotective excitatory amino acid receptor antagonist, kynurenic acid, was examined in the rat in six brain regions and in the liver. KAT activity increased in all brain areas (but not in the liver) between 3 days and 3 months post-natum, and substantial differences were observed in the rates of the increase. For example, KAT activity in the parietal cortex increased 34 fold during the observation period, whereas enzyme activity in the cerebellum and substantia nigra increased only 3-5 fold over the same interval. Intermediate developmental changes were found in striatum, hippocampus and olfactory bulb. The differential increases in KAT activity became particularly pronounced after the second postnatal week. Since N-methyl-D-aspartate (NMDA) receptors, which are preferential targets of kynurenic acid, play an important role in brain development, the regional heterogeneity of KAT's ontogenetic profile may have a bearing on the function and dysfunction of the young central nervous system. PMID- 8403388 TI - Developmental expression of the growth hormone receptor gene in the rat hypothalamus. AB - The ontogeny of growth hormone receptor (GHR) gene expression was studied in the rat hypothalamus. Total RNA from the hypothalamus of rats at different developmental stages (embryonic day 15-56 days of age) was characterized using a 32P-labeled RNA probe derived from the extracellular domain of the rat GHR cDNA. Two RNA species, 4.5 kilobases (kb) encoding for GHR and 1.2 kb encoding for GH binding protein, were detected in hypothalamic tissue from embryonic day 15 to 56 days of age. Their levels were low at embryonic day 15 and increased toward 3 days of age. The level of 4.5-kb transcript preferentially increased from 7 days after birth, and it was maintained until 35 days of age. Thereafter, the level of 4.5-kb transcript declined. The ratio between the 4.5- and 1.2-kb transcripts was less than 2.0 from embryonic day 15 to 3 days after birth, while it was larger than 4 after 7 days of age. There was no sex difference in the levels or the ratios of the transcripts of the GHR gene from 7 to 56 days of age. The findings indicate that the 4.5-kb transcript preferentially processed postnatally in the rat hypothalamus. PMID- 8403389 TI - Urinary selenium concentrations. AB - Urinary selenium concentrations are used as an indicator of selenium status. A strong correlation has been established between dietary selenium and daily urinary selenium excretion in a wide range of populations from all over the world with different dietary selenium intake. Data on urinary selenium concentrations in healthy individuals and patients with different pathological conditions are reviewed. Selenium excretion rates of 20-200 micrograms/day are not associated with deficiency or toxicity problems. Urinary Se excretion is decreased in children, elderly people, and pregnant women. Workers exposed to heavy metals, and cancer patients, have higher and lower urinary Se concentrations, respectively, than control groups. The trimethylselenonium ion, a minor metabolite of Se in urine, assumes a significant role only in the detoxification of excess Se intake. Studies of bioavailability and balance show the important role of the kidneys in homeostatic regulation of Se. PMID- 8403390 TI - Detection and characterization of blocking-type anti-acetylcholine receptor antibodies in sera from patients with myasthenia gravis. AB - We developed a highly sensitive, convenient assay for measuring blocking-type anti-acetylcholine receptor antibodies, which inhibit the binding of 125I-labeled alpha-bungarotoxin (alpha-BuTx) to the acetylcholine receptor (AChR). This procedure detected inhibitory activities in sera from 76% of patients with myasthenia gravis. Results of an experiment done with synthetic peptide corresponding to the alpha-BuTx binding region in the alpha-subunit of Torpedo AChR suggested that this inhibition is due to nonspecific steric hindrance caused by the binding of antibodies to a region other than the alpha-BuTx site, rather than by direct binding to the latter site. The inhibitory activities of the blocking-type antibodies and the titers of non-blocking-type antibodies were correlated. Moreover, the blocking-type antibodies could dissociate 125I-labeled alpha-BuTx from 125I-labeled alpha-BuTx-human AChR complex, and their dissociation activities showed good correlation with the inhibitory activities. PMID- 8403391 TI - Assay instrument-dependent matrix effects in standardization of cholesterol measurements. AB - Human serum-based frozen reference materials have been used by the Centers for Disease Control and Prevention (CDC)-National Heart, Lung and Blood Institute Lipid Standardization Program to improve the precision and accuracy of blood cholesterol measurements. Occasionally, laboratories in the program have had problems obtaining results for patients' fresh serum samples equivalent to those obtained with frozen CDC standardization pools. This incompatibility of sample, reagent, instrument, and assay characteristics has been labeled broadly as a "matrix effect," which usually is attributed to unknown characteristics of the processed pool material. In this study we showed that a large negative bias obtained with CDC pools was attributable to use of the sample blank mode on the Cobas-Bio analyzer. However, under the same conditions, fresh patients' serum samples were analyzed accurately. The use of a blank absorbance immediately after mixing sample and reagents (the "autoblank" mode) allowed the instrument to accurately analyze both fresh serum samples and CDC standardization pools and thus allowed the documentation of traceability of the cholesterol measurements to the National Reference System for Cholesterol. PMID- 8403392 TI - Assay of D-dimer based on immunofiltration and staining with gold colloids. AB - In this immunofiltration assay of D-dimer in plasma samples, the antigens are captured by a monoclonal antibody on a porous membrane, and labeled with the same antibody conjugated to gold colloids. The assay time is < 2 min, and a color of intensity proportional to the concentration of D-dimer is left on the membrane. The reference range (mean +/- 2 SD) was 0.336 +/- 0.133 mg/L (n = 69). Linearity was found up to 10 mg/L. Comparison with ELISA results (x) for 198 patients' samples demonstrated a linear regression equation of y = 0.99(+/- 0.05)x + 0.68(+/- 0.07) and a mean square error of 0.503. Comparison of visual reading of the color signal (y) vs reflectometric measurements (x) for 220 patients' samples demonstrated a linear regression equation of y = 2.5(+/- 0.06)x -0.22(+/- 0.04) and a mean square error of 0.095. Bilirubin, hemoglobin, fibrinogen, soluble fibrin, and fibrinogen degradation products and freezing/thawing of samples did not interfere. Some interference from rheumatoid factor, heparin, and the presence of cells or large lipid particles was seen. The variance (CV) was 8-12% within run, 10-18% between runs, and 13-20% between persons. The new assay constitutes a rapid and reliable analytical tool combining simplicity equivalent to that of latex tests with analytical information approaching that of ELISA. PMID- 8403393 TI - Interpretation of results for tumor markers on the basis of analytical imprecision and biological variation. AB - Interpretation of results for CA 15.3, carcinoembryonic antigen (CEA), and tissue polypeptide antigen (TPA) during breast cancer monitoring requires data on intra- (CVP) and inter- (CVG) individual biological variation, analytical imprecision (CVA), and indices of individuality. The average CVP and CVG obtained from 22 healthy women were, respectively, 6.2% and 62.9% (CA 15.3), 9.3% and 86.8% (CEA), and 28.3% and 133% (TPA). The indices of individuality were all < 0.6: 0.2 (CA 15.3), 0.15 (CEA), and 0.2 (TPA). CVA depended on the concentration of the analytes. CVP and CVA determine what constitutes a significant difference between sequential results. Assuming a CVA of 11.2% (CA 15.3), 9.5% (CEA), or 11.9% (TPA), results must differ by 30%, 31%, or 72%, respectively, for P < or = 0.05. We found that CVP and CVA contribute considerably to the variation during breast cancer monitoring. Consequently, both CVP and CVA should be considered in criteria for marker evaluation. Because of low indices of individuality, conventional cutoff limits are inappropriate both for initial identification and for follow-up of breast cancer. PMID- 8403394 TI - Passive hemagglutination inhibition test for diagnosis of brown recluse spider bite envenomation. AB - Our goal was to recreate a passive hemagglutination inhibition (PHAI) test to diagnose brown recluse spider (BRS; Loxosceles reclusa) bite envenomation for treatment trials. Guinea pigs received intradermal injections of concentrated spider venom from the following species: Loxosceles reclusa, Argiope aurantia, Argiope trifasciata, Phidippus audax, and Lycosa frondicola. Skin lesion exudate was collected and tested with the BRS venom PHAI assay. From 51 separate collections of exudate, test sensitivity was 90% as long as 3 days after venom injection. Specificity was 100% with venom from the other spider species listed above in vivo (7 test samples) and in vitro (5 test samples), as well as with random bacterial exudate with and without added serial dilutions of BRS venom (10 test samples). The test was reproducible over repetitive assays to within one 10 fold dilution. A positive PHAI test result could function as an entry criterion for BRS bite victims in human treatment trials. PMID- 8403395 TI - Carbohydrate-deficient transferrin quantified by HPLC to determine heavy consumption of alcohol. AB - We developed a new fully automated ion-exchange chromatographic method for quantitating carbohydrate-deficient transferrin (CDT) on a Mono Q column. Quantitation relies on the selective absorbance of the iron-transferrin complex at 460 nm. Transferrin isoforms deficient in sialic acid, with pIs 5.7 and 5.9, can easily be separated and quantitated as a percentage of the total transferrin. This method has been applied to samples from teetotalers, occasional drinkers, patients with recent heavy alcohol consumption, and patients during detoxification. The sensitivity of the method was 55% in patients reporting 40-70 g daily ethanol consumption and nearly 100% in heavily intoxicated patients (70 500 g daily consumption). The half-life of the dominating pI 5.7 isoform in this group was 9.5 (+/- 1) days during detoxification. A CDT value > 0.8% is a highly specific marker for alcohol abuse and is greatly superior to other currently available biological markers. PMID- 8403396 TI - Determination of apolipoprotein E genotypes by single-strand conformational polymorphism. AB - We used single-strand conformational polymorphism (SSCP) to determine apolipoprotein E (Apo E) genotypes in 47 individuals. A 295-base-pair (bp) DNA fragment coding for amino acid residues 80-178 of the Apo E protein gave distinct patterns for the three alleles. When we used SSCP to determine the Apo E polymorphism of five individuals whose phenotyping results differed from those of genotyping, the SSCP results agreed with the genotyping results obtained by the PCR-based amplification refractory mutation system (ARMS). Because most of the reported rare alleles of the Apo E gene involve mutations of amino acid residues in positions 120-160, our SSCP method is useful for determining rare as well as common alleles. PMID- 8403397 TI - Apolipoprotein E polymorphism in a healthy Swedish population: variation of allele frequency with age and relation to serum lipid concentrations. AB - We analyzed blood samples from 407 healthy Swedish individuals, 244 men and 163 women, ages 17 to 86 years, for apolipoprotein (apo) E isoforms and serum triglycerides, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and total cholesterol. Parallel genotyping by means of polymerase chain reaction (PCR)-amplified DNA was performed in 200 subjects. Identical results were obtained by genotyping and phenotyping in 95% of all subjects analyzed. The apo E allelic frequencies were 7.8% for epsilon 2, 71.9% for epsilon 3, and 20.3% for epsilon 4. Compared with other Caucasian populations, the present population had a high relative allelic frequency of epsilon 4. The epsilon 4 frequency decreased with increasing age and was significantly lower in individuals > 60 years of age (14.7%). When controlling for age and sex, there were strong correlations between total serum and LDL cholesterol and the various epsilon alleles. The epsilon 4 and epsilon 3 alleles correlated positively with serum cholesterol and the epsilon 4 allele correlated positively with LDL cholesterol. In contrast, HDL cholesterol and serum triglycerides did not show any correlation to the allele types. Thus, the results demonstrate a considerable age variation of the epsilon allele frequency among healthy Swedes and an influence of apo E alleles on serum and LDL cholesterol concentrations. PMID- 8403398 TI - Enzymatic rate assay of creatinine in serum and urine. AB - A two-step method for assaying creatinine in serum and urine samples, suitable with automated analyzers, is reported. Reagent 1, for the first step, contains a blanking system [creatine amidinohydrolase (CRTase), urease, glutamate dehydrogenase, NADPH, and 2-oxoglutarate] and a NADPH-regenerating system [Mg(2+) dependent isocitrate dehydrogenase (ICD), MgCl2, and excess isocitrate]. Reagent 2, for the second step, contains the metal-chelating reagent trans-1,2 cyclohexanediamine-N,N,N',N'-tetraacetic acid (CyDTA) and a trigger system [creatinine amidohydrolase (CRNase)]. When a specimen is mixed with reagent 1, all the creatine, urea, and NH3 present are removed by the blanking and NADPH systems. On adding reagent 2, CyDTA inactivates ICD to inhibit the NADPH system. Simultaneously, the creatinine (1 mol) in the specimen is hydrolyzed into creatine by CRNase, and then releases NADP+ (2 mol) through the blanking system. Our optimized method can determine creatinine linearly up to 500 mg/L, with within-day CVs < 1.2% and day-to-day CVs < 2.7%. PMID- 8403399 TI - Enzyme immunoassay, kinetic microparticle immunoassay, radioimmunoassay, and fluorescence polarization immunoassay compared for drugs-of-abuse screening. AB - The newest formulation of the Syva EMIT assay for drugs of abuse, EMIT II, and a new immunoassay, OnLine (Roche), utilizing the kinetic interaction of microparticles in solution (KIMS) methodology, RIA tests, and TDx fluorescence polarization immunoassay (FPIA) procedures were compared for marijuana, cocaine, opiates, and barbiturates. Both EMIT II and OnLine immunoassays were performed with a Hitachi 717 analyzer. Calibration curves, the degree of separation between negative and cutoff calibrators, precision, likelihood of carryover from positive to negative samples, and overall ease and speed of analysis were evaluated. RIA and OnLine detected 99% of gas chromatography/mass spectrometry (GC/MS)-confirmed marijuana samples; TDx, 95%; and EMIT II, 88%. All four immunoassays detected approximately 99% of confirmed cocaine-positive urines. RIA, OnLine, and TDx all detected 100% of opiate-confirmed samples; EMIT II, 97%. Barbiturate assays exhibited the greatest disparity, with OnLine and TDx detecting 100% of confirmed positives; EMIT II, 88%; and RIA, 78%. For a variety of reasons, we prefer the fully automated EMIT II and OnLine assays for high-volume urine testing, in comparison with our laboratory's semiautomated RIA tests and the limited throughput TDx system. The four immunoassays investigated delivered comparable performance in terms of detection rates for GC/MS-confirmed positives for some drugs but not for others. PMID- 8403400 TI - Highly sensitive enzyme immunoassay of proinsulin immunoreactivity with use of two monoclonal antibodies. AB - A highly sensitive two-site sandwich ELISA measuring total proinsulin immunoreactive material in serum or plasma was developed. The assay was based on two monoclonal antibodies, an anti-C-peptide antibody bound to a microtest plate and a biotin-labeled anti-insulin antibody. The detection limit (3 SD above zero value) in buffer was 0.05 pmol/L, corresponding to 0.25 pmol/L in human serum (diluted 1:5). The linear calibrator range was 0.05-20 pmol/L. Interassay CVs were 4.7% at a median (range) of 2.3 pmol/L (1.4-2.8 pmol/L, n = 8), 6.7% at 5.1 pmol/L (3.3-8.0 pmol/L, n = 8), and 8.7% at 10.0 pmol/L (8-12 pmol/L, n = 10). Mean analytical recovery of added human proinsulin (hPI) (2, 5, and 10 pmol/L) to serum was 84% (range 68-128%, n = 9). Human insulin and human C-peptide did not cross-react at 5000 and 10,000 pmol/L, respectively. The four major proinsulin conversion intermediates reacted 65-99%: split(32-33)hPI 74%, des-(31,32)hPI 65%, split(65-66)hPI 78%, and des(64,65)hPI 99%. All serum values from 38 fasting healthy subjects were above the detection limit: median (range) 4.0 (2.1-12.6) pmol/L. PMID- 8403401 TI - Competitive time-resolved immunofluorometric assay for quantifying carbonic anhydrase VI in saliva. AB - A competitive time-resolved immunofluorometric assay sensitive and robust enough for quantifying human salivary carbonic anhydrase isoenzyme VI (HCA VI) was developed. The solid-phase immunoassay is based on competition between Eu(3+) labeled HCA VI and salivary HCA VI for polyclonal rabbit anti-HCA VI antibodies that are attached to microtiter plate wells precoated with sheep anti-rabbit IgG. The subsequent immunoassay including the separation of free and bound HCA VI requires only one incubation step, after which the Eu3+ of the bound labeled antigen is released into an enhancement solution. The highly fluorescent Eu chelates formed in this solution are then quantified by time-resolved fluorometry (Delfia). The time-resolution principle effectively obviates possible interferences from complex biological material such as saliva. The assay detection limit was 1.5 micrograms/L. Intra- and interassay imprecisions (CVs) were 5.1% and 5.3%, respectively. The mean analytical recovery was 93%. The mean +/- SD concentration of HCA VI in paraffin-stimulated saliva was 6.8 +/- 4.3 mg/L (n = 30) and the secretion rate was 10.2 +/- 7.9 micrograms/min. The method was useful for further investigations of the role of HCA VI in difficult matrices, e.g., saliva. PMID- 8403402 TI - Competitive assay to improve the specificity of detection of single-point mutations in alpha 1-antitrypsin deficiency. AB - Allele-specific oligonucleotides are used widely for the detection of single point mutations in genes. A modification of this assay based on competition has been developed for detection of the Z mutation of alpha 1-antitrypsin (alpha 1 AT). The normal alpha 1-AT allele is referred to as M, and the Z mutation arises from a single base substitution. Amplified DNA products corresponding to homozygous M, heterozygous MZ, and homozygous Z obtained by the polymerase chain reaction were incubated with a twofold molar excess of unlabeled oligonucleotide prior to hybridization with a radiolabeled oligonucleotide. Thus, initial incubation with unlabeled M-specific oligonucleotide was followed by hybridization with radiolabeled Z-specific oligonucleotide, and vice versa. This assay increased the specificity of single-point mutation detection three- to four fold. Furthermore, specific hybridization was obtained at a lower temperature as a consequence of improving the signal-to-noise ratio. PMID- 8403403 TI - Quantification of the cross-link pentosidine in serum from normal and uremic subjects. AB - Pentosidine is a fluorescent cross-link compound that accumulates in human tissues from uremic and diabetic patients. Using SP-Sephadex C-25 pretreatment before reversed-phase HPLC, we developed a method for quantifying pentosidine in the acid hydrolysate of serum. We examined concentrations of pentosidine in serum from 98 patients with end-stage renal disease requiring hemodialysis and from 33 normal control subjects. The mean (+/- SD) concentration of pentosidine was significantly greater in serum from uremic patients than that from control subjects (1267 +/- 695 nmol/L vs 77 +/- 40 nmol/L, P = 0.0001). There was a significant correlation between serum pentosidine concentrations and age in control subjects (r = 0.453, P < 0.01), but not in uremic patients. Serum pentosidine significantly increased with the duration of hemodialysis treatment (r = 0.272, P < 0.05). The greater pentosidine concentrations in uremic patients may be caused by the increased synthesis of pentosidine (perhaps because of the retention of pentosidine precursors), by the retention of pentosidine molecule itself in end-stage renal failure, or both. PMID- 8403404 TI - Chemiluminescent third-generation assay (Amerlite TSH-30) of thyroid-stimulating hormone in serum or plasma assessed. AB - We assessed the laboratory performance and clinical utility of a new commercial third-generation assay of thyroid-stimulating hormone (TSH), Amerlite TSH-30. The interassay CV was 6% at TSH concentrations of approximately 0.08 mIU/L, and the analytical and functional detection limits of the assay were 0.005 and 0.0125 mIU/L, respectively. Although the assay recovered approximately 96% of TSH International Reference Preparation (TSH-IRP) 80/558 added to serum samples, the endogenous TSH concentrations in basal samples were significantly lower than those found by using two other TSH assays; bias data obtained from thyroliberin stimulation tests suggested that the negative bias found with TSH-30 may be due to the heterogeneity of TSH in basal samples. TSH-30 completely discriminated hyperthyroid and hypothyroid patients from euthyroid ambulatory patients but also detected TSH (> 0.0125 mIU/L) in 3 of 46 untreated hyperthyroid patients. Compared with two second-generation assays, TSH-30 better discriminated between patients with subnormal TSH due to hyperthyroidism, thyroxine overreplacement, and nonthyroidal illness but there was still significant overlap between results for these groups. PMID- 8403405 TI - Kinetic determination of serum adenosine deaminase. AB - A new kinetic method for the determination of serum adenosine deaminase (EC 3.5.4.4) is described, with adenosine as the substrate and nucleoside phosphorylase and xanthine oxidase as the reaction enzymes. Inosine is produced, which is converted to hypoxanthine. The hypoxanthine is oxidized to xanthine, which is further oxidized to uric acid. In these two reactions, blue 2,6 dichlorophenolindophenol is reduced to a colorless compound and the decrease in color is measured spectrophotometrically at 606 nm. The assay was automated by using a Cobas Mira analyzer. The automated assay had a CV of < 7%, and the calibration curve was linear from 10 to 120 U/L. The assay correlates well with an established method, based on detection of liberated NH3 with Berthelot's reaction. The reference interval (mean +/- 2 SD) was 14-34 U/L (mean 24 U/L, n = 84). The enzymatic method described is easily automated and seems to be suitable for the routine determination of adenosine deaminase in serum. PMID- 8403406 TI - Dietary nutrient intakes and slight albuminuria in people at least 40 years old. AB - We studied the relation between diet and slight albuminuria in 5416 people, ages 40 years and over, who participated in a health screening survey of a local workforce. Degree of albuminuria showed log-linear univariate relations with dietary protein, cholesterol, and sodium intakes, and negative log-linear univariate relations with dietary fiber and polyunsaturated to saturated fat (P/S) ratio. After adjusting for age, gender, and ethnicity, the relative risk (95% confidence interval) for slight albuminuria was significantly increased in people reporting dietary cholesterol consumption > 226 mg/day compared with people reporting consumption < or = 226 mg/day [1.32 (1.02, 1.70)], and significantly reduced in people reporting dietary fiber consumption > 26 g/day compared with people reporting consumption < or = 26 g/day [0.74 (0.58, 0.95)]. There was no significant effect of dietary protein, P/S ratio, or salt intake. We conclude that risk of slight albuminuria is increased by consumption of dietary cholesterol and reduced by consumption of dietary fiber. PMID- 8403407 TI - Correcting for contamination from hemolysis in measurements of lactate dehydrogenase isoenzyme 1 in serum from patients with testicular germ cell. PMID- 8403408 TI - Automated measurement of mitochondrial aspartate aminotransferase by selective proteolysis with proteinase K. PMID- 8403409 TI - Methods for evaluation of protein components in bronchoalveolar lavage fluids. PMID- 8403410 TI - Determination of malonaldehyde in hyperlipemic sera. PMID- 8403411 TI - Nonlinearity of calcium measurement on the Kodak Ektachem at concentrations > 110 g/L (slide generations 18 and 19) PMID- 8403412 TI - Improved extraction, sensitivity, specificity, and stability for measuring fentanyl in plasma by gas chromatography with nitrogen detection. PMID- 8403413 TI - Increased serum creatine kinase activity after a mild hypothermic episode. PMID- 8403414 TI - Run-rejection survey to assess benefits of training in concepts of quality control and total quality management. PMID- 8403415 TI - Analytical inaccuracy for folic acid with a popular commercial vitamin B12/folate kit. PMID- 8403416 TI - Rapid glycohemoglobin method revisited. PMID- 8403417 TI - Automated enzymatic determination of inulin. PMID- 8403418 TI - Human chorionic gonadotropin as a thyroid stimulator: time for reassessment? PMID- 8403419 TI - Vanadate, epidermal growth factor and prostaglandin E2 production in human amnion cells. AB - We have investigated the effects of vanadate (VAN) and mouse epidermal growth factor (mEGF) on prostaglandin (PG) E2 production in human amnion cells in monolayer culture that served as a model system. The secretion of PGE2 into the culture medium was quantified by radioimmunoassay. The rate of conversion of [14C] arachidonic acid to [14C] PGE2 (PGE2 synthase) was determined in cell sonicates under optimal in vitro conditions. After a treatment period of 4 h we observed an increase in PGE2 production in the presence of mEGF (4.7-fold), VAN (1.8-fold) or both agents (6.8-fold) over control samples. Similarly, the specific activity of PGE2 synthase was stimulated maximally after 4 h with mEGF (2.1-fold), VAN (1.7-fold) or with both (4.8-fold) compared with untreated tissue samples. In the presence of cycloheximide those effects were abolished. We suggest, that VAN and mEGF act by a mechanism(s) that involves de novo protein synthesis or that alters the phosphorylation state of enzymes that are requisite for the conversion of arachidonic acid to PGE2. PMID- 8403420 TI - Functional analysis of tumor-associated lymphocytes from gynecological tumors. AB - Tumor-Associated Lymphocytes (TAL) were isolated from peritoneal fluids of six ovarian cancer patients and pleural effusion from eight breast cancer patients, respectively. In one case we obtained ascitic fluid as well as pleural effusion because of intraabdominal metastatic breast carcinoma. The collected cells were cultured in a complete medium and supplemented with human interleukin-2 (nIL-2) in a concentration of 1000 Units/ml. Phenotyping was not always possible due to rapid decay of the cells. Cytotoxicity was determined with a fluorescence-based assay, in some cases at different stages of cell growth. In two cases TAL from ascitic fluids showed increased cytotoxic activity after a longer cultivation period. TAL from pleural effusions showed cytotoxic activity against the target cell lines in two cases only. Some of these TAL did not proliferate any more but died within 24 h. With the functional analysis we wanted to investigate the cytotoxic potential against natural killer (NK)-sensitive and NK-resistant (Raji) cell lines. The results demonstrate the ability of some of the TAL populations to destroy tumor cells. PMID- 8403421 TI - The effect of pethidine hydrochloride on the cervical muscles in the active phase of labour. AB - The effect of pethidine hydrochloride (PHC) on the smooth muscles of cervix was studied in the active phase of induced labour in 27 primiparous women. The Electromyographic (EMG) root mean square (RMS) which provides information about the amplitude (intensity) of muscular cervical activity, and the median frequency (MF) which represents the EMG frequency spectrum were analyzed in connection with the intensity of intra-uterine pressure (IIUP). The RMS significantly decreases after the application of PHC, while the IIUP significantly increases. The effect of the applied drug disappears in one hour's time. The MF values increase insignificantly after the application and change in the opposite direction to the change of MSR. The change RMS, MF and IIUP values do not correlate with the duration of the active phase of labour. PMID- 8403422 TI - Ovarian cysts in the postmenopause: is a conservative treatment feasible? AB - The incidence of epithelial ovarian cancer reaches a peak between the ages of 50 and 59 years. Therefore any ovarian enlargement in the postmenopause has been treated up to now with prompt surgical exploration. Recently the reliability of ultrasound has allowed a conservative management of small unilocular ovarian cysts even in the postmenopause. The Authors report here their experience on benign masses in postmenopausal women, and discuss the feasibility of ultrasound guided aspiration of small, anechoic adnexal cysts. PMID- 8403423 TI - The effects of vanadate and epidermal growth factor on the specific activities of phospholipase A2 and phosphatidylinositol-specific phospholipase C in human amnion cells. AB - We have evaluated the specific activity of phospholipase A2 (PLA2) and phosphatidylinositol-specific phospholipase C (PLC) in human amnion cells in monolayer culture that had not (CTL) or that had been treated with vanadate (VAN) and/or mouse epidermal growth factor (mEGF) for 4 h. It is known that both agents stimulate prostaglandin (PG) E2 synthesis in these cells. Phospholipase enzyme activities were determined in the 750 x g supernatant fraction of amnion cell homogenates under optimal in vitro conditions. The specific activity of PLA2 ranged from 1.1 to 1.25 nmol/mg protein/0.5 h and that of phosphatidylinositol specific PLC from 1.04 to 1.2 mumol/mg protein/h. Treatment of amnion cell cultures with VAN and/or mEGF had no statistically significant effect on the specific activities of either phospholipases. Thus, we conclude that the stimulation of PGE2 production by VAN and mEGF is not due to an increase in the release of arachidonic acid from glycerophospholipid storage forms in human amnion cells. PMID- 8403424 TI - The role of steroid hormones in cervical ripening. AB - Dehydroepiandrosterone sulphate (DHEA-S), estradiol (E2), estriol (E3), progesterone (P4) and cortisol (F) maternal plasma concentrations were determined using enzyme or fluoroimmunoassays in 28 healthy pregnant women prior to the induction of labour, performed for various reasons, in order to evaluate their impact on cervical ripening. Mean maternal and gestational ages were 24.7 +/- 3.6 years and 39.1 +/- 1.9 weeks, respectively, and the gravidity ranged from 0-4. The subjects were divided according to cervical maturity into Group A (n = 13, Bishop scores > 4) and Group B (n = 15, Bishop scores < 4). There were no statistically significant differences observed between the two groups as to concentrations of oestradiol, oestriol, progesterone and cortisol. However, significantly higher concentrations of DHEA-S were found in group A compared to group B (1335 +/- 885 ng/ml and 558.2 +/- 191.4 ng/ml, respectively), (p < 0.001). In conclusion, the high DHEA-S concentrations in maternal plasma may play an important role in pregnancy by producing favourable cervical conditions for delivery or by triggering the labour itself. PMID- 8403425 TI - Uterine fibroids: protocols of integrated medical/surgical treatment. AB - Uterine fibroids are the most common solid pelvic tumor in women and in these last years their management has been deeply reviewed. An optimal integration of GnRH analogue treatment with different surgical techniques require an adequate work-up for the specific problems of each single patient. For a correct management of these patients, we divided them into 4 categories on the basis of symptoms and specific objectives to persuasive: a) perimenopausal patients; b) young patients, symptomatic or with large myomas, with no wish for more children; c) young patients, symptomatic or with large myomas, wishing to preserve fertility; d) infertile patients or patients with history of repeated miscarriages. Our suggested work-up is the following: a careful USGraphic evaluation of fibroid size and localization, a transvaginal doppler examination of the uterine blood supply, a complete haematochemical analysis, a hysteroscopy guided biopsy and a complete bone evaluation. On the basis of the above mentioned evaluations for each group we will use a different therapeutic approach in regard of either length of medical treatment (for tree to six months, for one or more cycles) or necessity of surgical treatment (with the possibility to get a natural menopausal), or different surgical techniques (operative hysteroscopy and/or laparoscopy, laparotomic myomectomy, vaginal or abdominal hysterectomy. PMID- 8403426 TI - The evolution of the management of "CIN" lesions. AB - During the years 1961-1986 a total of 378 CIN lesions were diagnosed among 4875 women screened for cervical pathology; 285 of them were followed-up for a minimum of 5 years. All 169 cases diagnosed as CIN-3 and 18 classified as CIN-2 were treated surgically, whereas 89 patients diagnosed as CIN-1 and 65 classified as CIN-2 were treated conservatively. Finally, 37 cases (23 diagnosed as CIN-1 and 14 diagnosed as CIN-2) did not receive any treatment. Of the 187 women treated surgically, 6 (3.8%) showed, within the five year follow-up period, progression or recurrence of the disease, which necessitated either additional surgery or radiation; 5 of these cases had been classified as severe dysplasia (CIN-3). Recurrence or progression necessitating major surgery, occurred in 8 (5.1%) of the 154 patients treated conservatively; 5 of these cases were classified as moderate dysplasia (CIN-2). The data indicate that present diagnostic procedures allow a better selection of patients to be submitted to surgery, increasing the proportion of those treated conservatively, without jeopardizing their prognosis. PMID- 8403427 TI - A subcutaneous uterus with unusual presenting features. AB - A 32 year old female, para 2 + 0 presented with a hard lump in the scar of a lower midline incision. She had had a myomectomy 2 years previously and subsequently noticed the lump 3 months later. Her only complaints were urinary frequency during menstruation and the suprapubic mass. Surgery was performed for what was initially thought to be a desmoid tumour. At surgery the uterus was found to be lying in the subcutaneous position with no peritoneal sac. The uterus was dissected free of the sheath and reduced into the pelvis, uneventfully. This rare occurrence of a subcutaneous non-gravid uterus in the absence of a hernial sac is reported and its clinical features and possible preventative measures are discussed. PMID- 8403428 TI - Progesterone immunosuppressive levels and luteal steroid profiles in the cycles induced with clomiphene citrate. AB - Several studies have focused their attention on the possible interferences of the endocrine with the immunity system; these interrelations have been summoned to explain some aspects of the implantation. It has already been demonstrated that progesterone could play an immunosuppressive role, allowing the implantation of allotransplantation. However, the individual plasmatic levels of the substance needed to produce that effect still unknown. With the aim of determining the certain immunosuppressive progesterone levels in women with normal ovulatory function, and in order to determine if the other principal steroids might also have immunosuppressive effects, in 47 women affected by sine causa infertility, treated with 100 mg. die of Clomiphene Citrate (from day 3 to day 7) we evaluated the plasma levels of progesterone, 17-OH-Progesterone and 17 beta-Estradiol. The assays were made on the 7th, 11th and 14th post-ovulation days both in women who conceived (immunosuppressive effect present) and in women who did not achieve pregnancy (immunosuppressive effect absent). The results achieved showed a significant difference only in the progesterone values, while those of the other steroids were not significantly different, indicating thus that progesterone is the main element responsible for the immunosuppressive phenomenon and that the seriated evaluation in the luteal phase of this steroid could be used as a marker of achieved implantation. PMID- 8403429 TI - The estimation of fructosamine and HbAlc in pregnant women with diabetes mellitus. AB - Fructosamine, HbAlc, glucose, albumins and total proteins were estimated in 40 healthy pregnant women and 80 pregnant women with insulin dependent diabetes mellitus. Fructosamine was estimated by the NBT method with "Fructosamine test" commercially available kit on Technicom automatic analyser RA-1000. Glucose was determined on Beckman glucose analyser. HbAlc was assayed by the Bio-Rad test, while albumin and total proteins by Beckman tests. For all estimated parameters no significant differences were found between healthy pregnant women and pregnant women with insulin dependent diabetes mellitus. PMID- 8403430 TI - Molecular weight and substrate characteristics of human serum arylesterase following purification by immuno-affinity chromatography. AB - Human serum arylesterase (EC 3.1.1.2) was purified over 400-fold with a recovery of 61-78% by single-step immuno-affinity chromatography using a monoclonal antibody, 12-1H8, as the adsorbent. The resultant preparation behaved as one component on SDS-PAGE with an apparent M(r) of 46,000, indicating its high homogeneity. Molecular weight was determined as 380,000 Da by HPLC on TSK-gel G 3000SW. The arylesterase molecule would thus appear to be comprised of octameric subunits. The purified protein hydrolyzed paraoxon. The present study suggests that a separate classification of EC 3.1.8.1 for its paraoxon hydrolyzing activity may not be appropriate to differentiate from arylesterase activity. In addition, the substrate for arylesterase is discussed in terms of its chemical structure. PMID- 8403431 TI - A method for the assay of phosphatidylinositol-specific phospholipase D activity in serum. AB - A reproducible substrate for the assay of phosphatidylinositol-specific phospholipase D (PIPLD) can be prepared by extracting alkaline phosphatase from placental tissue with n-butanol under alkaline conditions. The alkaline phosphatase thus prepared retains its hydrophobic glycan phosphatidylinositol (GPI) anchor and aggregates into high M(r) forms. Incubation with serum hydrolyses the phosphate inositol linkage by PIPLD action, producing a less lipophilic, non-aggregated isoform of alkaline phosphatase. Three methods of measuring the amount of this isoform produced after a timed incubation with serum are described and compared: two types of phase partitioning systems, and electrophoresis and densitometry of the products after gradient-pore electrophoresis. All give comparable and reproducible measurements of PIPLD; however, the electrophoretic method is preferred for routine analysis. PMID- 8403432 TI - Survey of methods for measuring human anti-mouse antibodies. HAMA Survey Group. AB - Human anti-mouse antibodies (HAMA) methods were surveyed using a panel of 17 serum and two plasma specimens which were circulated to ten laboratories for analysis. The panel included HAMA-negative and HAMA-positive specimens (from patients given a variety of mouse monoclonal antibodies) and a specimen with a high rheumatoid factor titer. The participating laboratories used a diverse range of methods to determine HAMA and all three of the commercially available quantitative methods for HAMA were represented. The survey revealed significant differences in HAMA results among the different laboratories. PMID- 8403433 TI - Enzymatic diagnosis of steroid sulfatase deficiency by high performance liquid chromatography. AB - We established a reversed phase high performance liquid chromatographic (HPLC) method of assaying lymphocyte steroid sulfatase activity using estrone sulfate as the substrate. Application of this method for diagnosis of 8 patients allowed us to clearly distinguish the patients from the normal controls. This method is simpler and less expensive than the method previously reported, since neither radioisotope labeled substrates nor radioisotope facilities are required. We consider it to be easily used and widely available in most clinical laboratories. PMID- 8403434 TI - Glycan microheterogeneity of alpha 1-antitrypsin in serum and meconium from normal and cystic fibrosis patients by crossed immunoaffinoelectrophoresis with different lectins (Con A, LCA, WGA). AB - In order to test whether abnormalities of glycosylation occur in cystic fibrosis (CF), the glycan microheterogeneity of alpha 1-antitrypsin (alpha 1-AT) was studied in serum and meconium from normal individuals and patients with cystic fibrosis, by crossed immunoaffinoelectrophoresis (CIAE) using free Concanavalin A (Con A), Lens culinaris lectin (LCA) and wheat germ agglutinin (WGA). Three main results emerged from this study: (1) modification of glycosylation in serum alpha 1-AT from patients with cystic fibrosis were only significant with free Con A and WGA; this probably results from a reduced synthesis of the bi-antennary side chains or by their increased catabolism. (2) Differences in isoforms found in alpha 1-AT from normal individuals and patients with CF using free Con A, LCA, were more pronounced in the meconium than in the serum; this may provide a useful test in diagnosis of cystic fibrosis. (3) There was parallelism between the behaviour of alpha 1-AT in serum and meconium from patients with CF using LCA, Con A; this may be explained by different types or levels of disfunction affecting a glycosylation mechanism. PMID- 8403435 TI - Vitamin E attenuates induction of elastase-like activity by tumor necrosis factor alpha, cholestan-3 beta,5 alpha,6 beta-triol and linoleic acid in cultured endothelial cells. AB - Disturbances in arterial wall elastin metabolism appear to be important factors in atherosclerosis development. To evaluate this hypothesis, elastase-like activity was determined in cultured endothelial cells and their surrounding media after exposure to tumor necrosis factor-alpha (TNF), cholestan-3 beta,5 alpha,6 beta-triol (Triol) and linoleic acid (18:2). Significant increases in elastase like activity both in the cells and in the media were observed when subconfluent endothelial cells were treated with 12 microM Triol, 500 U TNF/ml, or 90 microM 18:2, for 72 h in the presence of 5% calf serum. Even higher activities were measured when endothelial cells were seeded directly into media enriched with 18:2, TNF or Triol and treated for 72 h. Vitamin E supplementation (25 microM) attenuated elastase-like activity in cells and media, independent of treatment. These results suggest that elastase-like enzyme induction in endothelial cells may be involved in cellular perturbations induced by certain lipids and cytokines. Vitamin E may provide a protective function by preventing the induction of elastolytic enzymes. This may have implications in elastin metabolism and atherosclerosis. PMID- 8403436 TI - Changes in immunoreactive manganese-superoxide dismutase concentration in human serum after 93 h strenuous physical exercise. PMID- 8403437 TI - Cigarette-smoking, birthweight, thiocyanate and fluorescent lipid-peroxidation products in maternal and cord plasma. PMID- 8403438 TI - Measurement of histidase activity in human fingernail by reversed-phase HPLC. PMID- 8403439 TI - Effects of ethanol on the assay of acyl-CoA:cholesterol acyltransferase (ACAT) PMID- 8403440 TI - Quantification of alpha rhythm desynchronization using the acceleration spectrum entropy of the EEG. AB - Desynchronization of the alpha rhythm is usually quantified as a loss of power in the alpha band. In this study we describe an alternative approach where desynchronization is characterized as an increase in the irregularity of the EEG signal. A new measure, the acceleration spectrum entropy (ASE), is used to quantify EEG irregularity. The ASE is the normalized information entropy of the amplitude spectrum of the second derivative of a time series. The ASE is a measure of the randomness or irregularity of a time series and ranges from 0 (straight line) to 100 (discrete random signal). The ASE was calculated for short EEG epochs under eyes closed and eyes open conditions in 15 subjects. There was a significant increase in ASE in the eyes open condition, reflecting an increase in EEG complexity. The ASE is a sensitive measure of EEG desynchronization. PMID- 8403441 TI - The influence of light drowsiness on the latency and amplitude of P300. AB - Light drowsiness affected the P300 obtained with a standard auditory oddball paradigm. A simple two-tone discrimination paradigm was used. The frequent nontarget stimuli were 1000 Hz pure tones and the infrequent target stimuli were 2000 Hz pure tones. The subjects were required to press a button whenever infrequent target tones were presented. With light drowsiness, P300 increased in latency and decreased in amplitude, but the counts of infrequent tones remained correct nevertheless. It was concluded that electrophysiological brain function differs in the light drowsy and awake states. In studies on P300, it is essential to rule out light drowsiness to obtain valid P300 amplitudes and latencies. PMID- 8403442 TI - Trifocal independent periodic lateralized epileptiform discharges. AB - The EEG of a 2 1/2-year-old female with a severe cerebral insult showed trifocal periodic lateralized epileptiform discharges (TRIPLEDs). We believe this is the first such record reported. As with BIPLEDs, the physical findings associated with this EEG appear to represent a diffuse cerebral insult. The most frequent causes of BIPLEDs are diffuse anoxic encephalopathy and CNS infection. In this patient, the TRIPLEDs are apparently due to a severe anoxic insult suffered in utero. As is true with BIPLEDs, the presence of TRIPLEDs represents a poor prognosis. PMID- 8403443 TI - Treatment resistant depression in a case of minor head injury: an electrophysiological hypothesis. AB - A relatively small but highly concordant literature suggests that manic depressive psychoses may include familial as well as nonfamilial subtypes. The latter, which appears to be an acquired form, follows brain injury of various etiology, displays EEG abnormalities and tends to respond well to anticonvulsant therapy. In this study we postulate an extension of this dichotomy to a larger spectrum of affective disorder, including milder but "treatment resistant" forms often associated with a high degree of dysfunction. Central to this hypothesis is information gathered from the longitudinal study of a well defined case in which precise clinical and electrophysiological data have been obtained at critical junctures. This data also leads us to suggest the existence of a latent vulnerability to psychosocial stressors in a subgroup of minor head injured patients. Once triggered, the resulting psychopathological state may be clinically indistinguishable from similar but etiologically distinct conditions. However, they respond poorly, if at all, to the treatments usually effective for mood disorders, often causing puzzlement and frustration among clinicians as well as mounting hopelessness in patients. This organic mood disorder subtype, which can be described as "neuro-sensitization mood disorder," may be identified by combining a thorough history, including perinatal events and putative brain injury, with electrophysiological data consisting of quantitative EEG (QEEG) in association with evoked potentials. In cases with positive findings, anticonvulsants such as carbamazepine, clonazepam and valproic acid can be a treatment of choice. PMID- 8403444 TI - The topography of muscle activity in quantitative EEG. AB - Ten normal preadolescent subjects were studied on three occasions with quantitative EEG topography: two sessions recorded EEG that was free of artifact, but during the third the subjects were instructed to clench their teeth and tighten their faces to produce muscle artifact. The sessions were then compared for stability of various frequencies at standard scalp electrode sites. The posterior electrodes were stable among sessions for frequencies up to 24 Hz; the anterior electrodes were less stable, and above 24 Hz there were no stable electrode sites. Muscle artifact contaminates anterior electrode sites more than posterior sites, making the posterior scalp electrodes superior for studying beta activity in quantitative EEG. Frequencies above 24 Hz are contaminated at all sites and therefore cannot be assessed reliably in the presence of muscle artifact. PMID- 8403445 TI - EEG, quantitative EEG, BAEP and ERP in centenarians. AB - In 11 healthy centenarians, the principal EEG and qEEG abnormalities were background frequency slower and power lower than in younger adults. The frequency and amplitude of anterior and posterior head regions on the EEG showed no difference, but qEEG demonstrated that posterior was higher than anterior. In 5 of 11 persons the frequencies of the temporal lobe were asymmetric. BAEP and ERP were measured in 6 persons. The latencies of I, III waves of BAEP were prolonged in only 1 subject. The latencies of exogenous components (P80, N100) of ERP were in the normal range. The latencies of endogenous components (N200, P300) of these 6 persons were prolonged, but in 1 person the N200 was normal and in 2 the P300 was normal. PMID- 8403446 TI - Topographic EEG analysis in two patients with basilar thrombosis. AB - The EEG activity of two patients with occlusion of the vertebral and basilar arteries was analyzed. Power maps, peak power frequency, and field power differences during both photic drive and verbal command were evaluated. Clinical findings of one patient fit the criteria of locked-in syndrome. Photic stimulation and event related desynchronization paradigm was used for testing the reactivity of EEG. Averaged EEG epochs during intended movement after verbal command showed significant alpha and sub-alpha power reduction. Regional differences of EEG reactivity were assumed secondary to the underlying hemispheric infarcts. Awareness of the patient in a locked-in state was documented by EEG analysis. In the second case permanent vegetative state was associated with a nonreactive rhythmic alpha pattern. Time sequence analysis of power ratios showed spontaneous alternating activity of alpha and sub-alpha generators. The authors conclude that pseudo-periodic fluctuation of alpha activity reflects partial preservation of thalamo-cortical connections. PMID- 8403447 TI - Triphasic waves in myxedema coma. AB - Triphasic waves occur in metabolic and nonmetabolic encephalopathies. We report an elderly patient in whom triphasic waves were associated with myxedema coma and disappeared after thyroid replacement therapy was initiated. The association between myxedema coma and triphasic waves has not been previously reported. PMID- 8403448 TI - Familial bilateral antecubital pterygia with severe renal involvement in nail patella syndrome. AB - A family in whom several members are affected with nail-patella dysplasia is reported because of severe renal involvement and bilateral antecubital pterygia. The family presented as arthrogryposis because of the elbow contractures. PMID- 8403449 TI - Congenital contractural arachnodactyly in two double second cousins: possible homozygosity. AB - A Bedouin family with two girls affected by severe congenital contractural arachnodactyly (CCA) is described. The girls were double second cousins. One of the girls also had ambiguous genitalia, an anomaly not generally associated with this disorder. The two children were both the product of first-cousin Bedouin parents from the same family. It is possible that both sets of parents were heterozygous for CCA; thus the infants may have been homozygous for CCA, which is usually an autosomal dominant condition. No instance of homozygous CCA has previously been reported. This family suggests genetic heterogeneity in CCA and that, in some rare families, the mode of inheritance may be autosomal recessive. PMID- 8403450 TI - Craniofacial anthropometric studies in Waardenburg syndrome type I. AB - A set of 15 surface measurements taken directly from the craniofacial region were determined in 51 patients with Waardenburg syndrome type I (WSI). A roentgencephamometric analysis including 20 linear dimensions was also performed by the use of cephalograms of 28 patients. Pairs between patients and controls of the same sex and age were established for comparison. The head circumference, clivus length and facial depth were smaller in affected individuals. The patients also had narrow nose, marked hypoplasia of the nasal bone, short philtrum, and short and retropositioned maxilla. A discriminant analysis revealed that the inner intercanthal distance, philtrum length, lower facial height and nasal bone length were excellent discriminating parameters of WSI. PMID- 8403451 TI - Ring chromosome 20 with loss of telomeric sequences detected by multicolour PRINS. AB - A ring chromosome 20 in a male infant with epileptic seizures, mental and somatic growth retardation, and behavioural disturbances is described. Conventional cytogenetics revealed the karyotype to be 46,XY,r(20)(pter-->qter) and no signs of mosaicism were found. Fluorescence in situ hybridisation using the clone p20Z1 identified the ring to be derived from chromosome 20. By counting 111 metaphases, only 7% were found to be missing the ring. The absence of telomeric sequences in the ring chromosome was demonstrated by multicolour PRINS: a three-step PRimed IN Situ labelling technique, using unlabelled primers. A terminal deletion of both arms thus seems to be the cause of the ring formation in the proband. Bivariate flow-analysis of chromosomes verified a deletion of the ring chromosome. The clinical and cytogenetic findings are compared with previous cases. A specific ring 20 syndrome seems justified. PMID- 8403452 TI - A family study on isolated congenital radial and tibial deficiencies in Hungary, 1975-1984. AB - Radial and tibial deficiencies are frequently (70%) associated with non-limb abnormalities. Isolated radial and tibial deficiencies may have a different etiology: in this study radial deficiencies were more frequent, there were milder subtypes and one-limb involvement was found in 70% of cases, tibial deficiencies were rare, mild subtypes did not occur and in general more limbs were involved. Among multimelic cases, one case had hypoplasia of the tibia with polydactyly and two cases had tibial aplasia with split hand +/- foot of autosomal dominant inheritance. Of 40 cases, four were familial. Findings of a case-control study on this population-based and validated 10-year cohort showed a lower mean birth weight and a higher rate of low birth weight in cases with isolated radial and tibial deficiency. PMID- 8403453 TI - Proposed: an international code of ethics for medical genetics. PMID- 8403454 TI - Frontonasal malformation and reciprocal translocation t(15;22)(q22;q13) PMID- 8403455 TI - Phenotypic intrafamilial heterogeneity in cystic fibrosis. PMID- 8403456 TI - The breakpoints of the EEC syndrome (ectrodactyly, ectodermal dysplasia and cleft lip/palate) confirmed to 7q11.21 and 9p12 by fluorescence in situ hybridization. PMID- 8403457 TI - Malignant lymphoma in a Bloom's syndrome patient treated with insulin. PMID- 8403458 TI - Second observation of Silver-Russel syndrome in a carrier of a reciprocal translocation with one breakpoint at site 17q25. PMID- 8403459 TI - Prenatal diagnosis of congenital sialidosis. AB - A case of prenatally diagnosed congenital sialidosis is described in a 21-week old male fetus, which was the fifth product of non-consanguineous parents. The proband, the second product, was diagnosed as having sialidosis by the enzyme assay in peripheral leukocytes after birth. At the 17th week of pregnancy, the fetus at risk was proven to have isolated sialidase deficiency after analyzing a sample of the cultured amniotic fluid cells. There were many cytoplasmic vacuoles and increased amounts of sialyloligosaccharides in the tissue of the aborted fetus, while the amount and the pattern of gangliosides in the central nervous system were normal. PMID- 8403460 TI - A trial of accelerated PUVA in psoriasis. AB - Ten psoriatic patients were treated with an accelerated PUVA regimen. Nine patients cleared in an average of 13.7 treatments (range 4-24) after an average total UVA dose of 83 J/cm2 (range 18-186). One patient had to withdraw due to pruritus and burning and six others had burning problems. The high incidence of burning may limit the routine use of this irradiation regimen. PMID- 8403461 TI - Skin microflora of atopic eczema in first time hospital attenders. AB - The bacterial flora of the skin was assessed quantitatively in 50 children with eczema, aged 6 months to 14 years, referred to the hospital for the first time. Twenty nonatopic controls with an unrelated non-infective disorder were also studied. Cotton-tipped swabs and contact agar discs were taken from the worst affected area of eczema and from an uninvolved site in patients and from the forearm in controls. Swabs were also taken from the nose, axilla and groin in all children. Bacterial colonization of the skin was consistently more common and greater in amount from patients compared with controls. Staphylococcus aureus was the most common pathogen isolated from patients only; from the worst affected area of eczema in 74% of patients and from an uninvolved skin site in 30% of patients. Quantitative assessment showed that the density of colonization was proportional to the severity of eczema. The most common S. aureus phage group was group II accounting for 32% of strains. Resistance to penicillin was present in 88% of strains and to two or more antibiotics in 38% of strains. No relationship was noted between the pattern of resistance and phage group. PMID- 8403462 TI - Protein S deficiency in a patient with necrotizing cellulitis. AB - A case of necrotizing cellulitis of the penis is reported. In this patient, transient functional protein S deficiency during the infectious process was demonstrated with a marked decrease of free protein S and elevated C4b-binding protein levels. Moreover, by investigating other members of this patient's family it was possible to demonstrate a subclinical inherited protein S deficiency. In this study it is suggested that the profound and transient decrease of functional protein S observed in this patient during infectious cellulitis, may have contributed to the pathogenesis of tissue necrosis. PMID- 8403463 TI - Immunohistopathological spectrum in cutaneous tuberculosis. AB - Granulomas of lupus vulgaris were characterized by a raised CD4+/CD8+ ratio, while in scrofuloderma this was significantly decreased. Tuberculosis verrucosa cutis showed intermediate changes. These immunological variations suggest that cutaneous tuberculosis forms a spectrum with strong cell-mediated immunity seen in lupus vulgaris, and weaker cell-mediated responses in scrofuloderma; tuberculosis verrucosa cutis is intermediate between the two. PMID- 8403464 TI - Effects of colchicine on the induction of ornithine decarboxylase and its gene expression by the phorbol ester tumour promoter. AB - The activity and gene expression of ornithine decarboxylase (ODC, an indicator of tumour promotion) were induced by the phorbol ester tumour promoter, 12-O tetradecanoylphorbol-13-acetate (TPA), in mouse skin. In the present study, the effect of colchicine, a microtubule-disrupting agent, on ODC activity and its gene expression were investigated. On administration of colchicine (100 micrograms) intraperitoneally 1.5 h before TPA treatment, ODC activity and ODC mRNA levels stimulated by TPA were suppressed to about 52 and 64%, respectively. These results suggest the involvement of a microtubule or colchicine-sensitive substrate in the signal transduction system for gene expression. PMID- 8403465 TI - Gamma/delta T-cell receptor expression in the jejunal epithelium of patients with dermatitis herpetiformis and coeliac disease. AB - The density of jejunal intra-epithelial T cells expressing the gamma/delta form of the T-cell receptor is known to be increased in coeliac disease, the significance of which remains a mystery. The expression of the gamma/delta T-cell receptor in the jejunum of patients with dermatitis herpetiformis, coeliac disease, treated and untreated, and controls were studied. Expression of the gamma/delta T-cell receptor was significantly increased in patients with dermatitis hepetiformis (P < 0.0005) and in both untreated (P < 0.0005) and treated coeliac patients (P < 0.05) compared with controls. There were significant correlations between the indices of enteropathy, enterocyte height (P < 0.005) and villous height/crypt depth ratio (P < 0.0001), and expression of the gamma/delta T-cell receptor in the jejunum of all the patients. This argues against the hypothesis that gamma/delta T-cells have a fundamental role in the aetiology of gluten-sensitive enteropathy. It suggests that gamma/delta T cells may be involved in the effector arm of the mucosal immune response to cereal peptides in susceptible individuals. PMID- 8403466 TI - Itraconazole in the treatment of tinea corporis and tinea cruris. AB - Forty-five patients with tinea corporis or tinea cruris were treated with oral itraconazole 100 mg daily for 15 days. At the end of the 15-day treatment, 80% of the patients were healed or had markedly improved. At the first follow-up visit, 2 weeks after stopping therapy, 80% of patients were considered responders. An additional follow-up visit another month later (i.e. 6 weeks post-treatment) showed that 32 of 41 patients had responded (78%). Overall, the mycological cure rate (culture and microscopy negative) was somewhat lower than the clinical response rate. Only three patients reported minor side effects (7%). Nausea was reported by two patients and an urticarial reaction was seen in one patient after 8 days treatment. This latter patient discontinued therapy because of the adverse experience. It is concluded that itraconazole, given at a daily dose of 100 mg for 15 days, is effective in the treatment of tinea corporis and tinea cruris. Response rates at the last visit (6 weeks post-therapy) remained at the same satisfactory levels as at the first follow-up visit (2 weeks post-therapy), even though treatment was stopped after 2 weeks. Itraconazole appears to be well tolerated by patients. These results, both in terms of efficacy and side effects, are in line with results reported by other investigators. The fact that the mycological cure rates were somewhat lower than the clinical response rates had apparently no influence on the relapse rate at 6 weeks follow-up post-therapy. PMID- 8403467 TI - The effect of essential fatty acids on epidermal atrophy due to topical steroids. AB - The effect of topical n-6 essential fatty acids in the form of evening primrose oil on the epidermal atrophy caused by a potent topical steroid was studied in 24 normal volunteers by measuring epidermal thickness and cross-sectional area and by histological examination. Epidermal thickness and cross-sectional area were significantly lower in normal forearm skin treated with 0.1% betamethasone valerate twice daily without occlusion for 3 weeks when compared with placebo treated skin. The addition of evening primrose oil to the topical steroid did not prevent steroid-induced epidermal atrophy suggesting that steroid-induced epidermal atrophy is not mediated by the inhibition of essential fatty acid release from cell membranes. PMID- 8403468 TI - Langerhans cell accumulation in chronic tinea pedis and pityriasis versicolor. AB - Persistence of chronic tinea pedis (CTP) and pityriasis (tinea) versicolor (PIVE) has been tentatively attributed to an impaired cellular immune response. Therefore immunophenotyping of the inflammatory infiltrates in both disorders was performed in order to detect possible defects in cellular defence. The results of the present study show a dominance of memory T cells, an accumulation of macrophages and lack of B cells. A very prominent feature in CTP and especially PIVE was a marked accumulation of Langerhans cells (LCs) in the epidermis, mostly without expression of CD4. Furthermore, reduced expression of cellular activation markers and the presence of suppressor T cells was noted. In general, the cellular response in PIVE appeared to be slightly stronger than in CTP. This composition of the inflammatory infiltrates in chronic dermatomycoses is similar to previous findings in a variety of dermatoses. However, in PIVE and CTP local reduction of delayed-type hypersensitivity may occur. The function of LCs in these infections is therefore of special interest. PMID- 8403469 TI - Widespread syringomata in Down's syndrome. AB - A case of widespread syringomata occurring in a woman with Down's syndrome is described. Whereas periorbital syringomata are common in Down's syndrome, more widespread syringomata have only rarely been reported. PMID- 8403470 TI - Linear lichen planus and lichen striatus--opposite ends of a spectrum. AB - The linear pattern of lichen planus (LP) is extremely rare. In six publications 1256 patients with LP have been reviewed and only three cases of linear LP (LLP) are reported. The differential diagnosis of LLP includes a number of linear dermatoses in particular lichen striatus (LS) and the extent of overlap between these syndromes remains contentious. PMID- 8403471 TI - Familial cold urticaria. AB - Familial cold urticaria (FCU) is a rare autosomal dominant condition, first described in 1940. The onset is in early life in all reported cases. Symptoms are triggered by generalized exposure to cold air, particularly in damp and windy weather. The cutaneous lesions consist of erythematous macules or plaques, urticarial lesions and sometimes petechiae. Associated fever, chills, joint pains, nausea, stiffness and swelling of the hands and feet frequently occur. The symptoms are variable, ranging from mild to incapacitating. The pathogenesis of FCU remains unknown. To our knowledge only 10 pedigrees have been published, seven from the USA and one each from Holland, France and South Africa. We wish to report another extensive pedigree after having had the opportunity to investigate one member of the family in detail. A short form of this pedigree has been published elsewhere. PMID- 8403472 TI - Contact sensitivity to lignocaine. AB - Lignocaine is a rare contact allergen, in contrast to other local anaesthetics such as benzocaine, which commonly sensitize. The case of a patient sensitized to lignocaine through the use of a topical medication for the treatment of haemorrhoids is reported. Previous reports show that application of a lignocaine containing preparation for pruritus ani is the most frequent cause of sensitization to lignocaine. The significance of sensitization to lignocaine is discussed in the light of its widespread use both in local anaesthesia and as an anti-arrhythmic. PMID- 8403473 TI - Degos disease and spastic paraplegia. AB - Malignant atrophic papulosis (Degos disease) is a rare disorder characterized by a vasculopathy of unknown origin. The cutaneous manifestations comprise erythematous papules, which heal to leave scars with a pathognomonic central porcelain-white atrophic area and a peripheral telangiectatic rim. There is usually involvement of the gastrointestinal tract but other organ systems can also be affected, the central nervous system being involved in 20% of cases. It is frequently fatal within 2 or 3 years from onset of systemic involvement, the cause of death usually being intestinal perforation. Our patient is of interest as she has survived an unusually long time despite florid cutaneous and neurological manifestations. PMID- 8403474 TI - UV-induced colloid milium. AB - A 39-year-old woman is reported who developed numerous confluent yellowish semitranslucent papules of the face over 8 months. The patient habitually exposed herself to UVA-radiation twice a week for 7 years for aesthetic reasons. Histological, histochemical and immunohistochemical examinations were consistent with the diagnosis of adult colloid milium. A review of the literature is discussed. This case emphasizes the close relationship between long-term sun exposure, solar elastosis and adult colloid milium. PMID- 8403475 TI - Primary cutaneous extramedullary plasmacytoma secreting lambda IgG. AB - An adult male was described, who developed a tumour on his left arm, which was diagnosed as a primary cutaneous extramedullary plasmacytoma after histopathological, electron microscopic and immunocytochemical studies. Further studies ruled out the presence of multiple myeloma, extramedullary plasmacytoma of another site, or paraproteinaemia/paraproteinuria. During a relapse of the tumour 2 1/2 years after initial treatment, a new immunocytochemical study demonstrated that the tumour cells expressed monoclonal lambda IgG at the intracytoplasmic level, an unusual finding. PMID- 8403476 TI - Fatal facial ulceration. AB - A case of an elderly man with a large facial ulcer is reported. The diagnosis of Wegener's granulomatosis was made at post-mortem when evidence of a granulomatous vasculitis was found in both the skin and lungs. This case serves to illustrate the difficulty in making the diagnosis of Wegener's granulomatosis, especially when it presents in its 'limited' form. PMID- 8403477 TI - Angioendotheliomatosis associated with Castleman's lymphoma and POEMS syndrome. AB - Castleman's lymphoma, a benign localized lymphoid hyperplasia, has been reported in association with POEMS syndrome, a multisystem reactive disorder that includes several skin manifestations. Benign reactive angioendotheliomatosis is a rare skin disorder that is associated with various systemic disorders. A patient with long-standing Castleman's lymphoma and POEMS syndrome also developed benign reactive angioendotheliomatosis. PMID- 8403478 TI - Dermatitis herpetiformis and primary sclerosing cholangitis. AB - A patient with dermatitis herpetiformis (DH) developed abnormal liver function tests and was found to have histological features of primary sclerosing cholangitis (PSC) on liver biopsy. The association of DH and PSC has not been previously reported and the high incidence of phenotype HLA B8 and DR3 in these disorders suggests a common genetic predisposition may exist for both diseases. PMID- 8403479 TI - Ulceration in necrobiosis lipoidica--a case report and study. AB - A case of aggressive ulceration of necrobiosis lipoidica was successfully treated with oral prednisolone. A retrospective study of 23 cases of necrobiosis lipoidica revealed a 13% incidence of ulceration. The pathogenesis, clinical features and treatment of ulceration in necrobiosis lipoidica are discussed. PMID- 8403480 TI - Cutaneous calcinosis--an unusual complication of intravenous phosphate administration. AB - The case of an 80-year-old woman who developed extensive cutaneous calcification following intravenous phosphate administration is presented. Also the circumstances under which cutaneous calcification may occur are discussed. PMID- 8403481 TI - Infectious mononucleosis presenting as erythema multiforme. AB - Infectious mononucleosis is rarely associated with skin lesions. A 23-year-old woman with acute infectious mononucleosis who presented with moderately severe erythema multiforme is reported. The role of steroids in the management of this condition is briefly discussed. PMID- 8403482 TI - Giant primary mucinous sweat gland carcinoma of the scalp. AB - Primary mucinous sweat gland carcinoma is a rare tumour which usually behaves in a relatively benign manner. This is illustrated by the massive proportions which this tumour reached in the case described, without evidence of metastases. PMID- 8403483 TI - Epidermolysis bullosa acquisita occurring in association with systemic lupus erythematosus. AB - A 77-year-old retired male physician with a 6-year history of systemic lupus erythematosus (SLE) developed a mechanobullous eruption, the features of which were clinically and immunopathologically consistent with a diagnosis of 'classical' epidermolysis bullosa acquisita (EBA). As EBA shares immunopathological findings with a number of cases reported as the 'bullous eruption of SLE', the clinical findings commonly recognized as 'classical EBA' may, in patients with SLE, represent a specific subset of the bullous eruption of SLE rather than a separate diagnostic entity. There are few reports in the literature describing classical EBA in patients with SLE. Findings in this patient add further support to the suggestion that EBA occurring in association with SLE, represents a subset of the bullous eruption of SLE, the clinical features of which may be modified by genetic susceptibility or disease activity. PMID- 8403484 TI - Lichen nitidus presenting as palmoplantar hyperkeratosis and nail dystrophy. AB - Four patients with lichen nitidus who presented with palmoplantar or nail lesions are reported. In three cases palmoplantar hyperkeratosis was marked; these cases also had nail-plate lesions, but in all four, lesions of lichen nitidus at other sites were absent, sparse or asymptomatic. PMID- 8403485 TI - Syringomas localized to the penis (case report). AB - Syringomas localized in the genital areas are unusual. There are only three cases reported describing syringomas on the penis. Owing to this atypical localization it is necessary to consider a differential diagnosis involving other papular lesions more frequent found on the penile skin. PMID- 8403486 TI - Kawasaki-like syndrome associated with griseofulvin treatment. PMID- 8403487 TI - Sticky palms--an unusual side-effect of etretinate therapy. PMID- 8403488 TI - The effect of a clear tar ointment on the minimal erythema dose for UVB. PMID- 8403489 TI - Follicular mucinosis associated with a Grawitz tumour. PMID- 8403490 TI - Striking the right balance; the role of cytokines in mycobacterial disease. PMID- 8403491 TI - Effect of anti-IL-4, interferon-gamma and an antifungal triazole (SCH 42427) in paracoccidioidomycosis: correlation of IgE levels with outcome. AB - Paracoccidioidomycosis is characterized by depressed cellular but enhanced humoral immune responses, which suggests a Th2 type of response to infection. We investigated possible therapeutic roles for anti-IL-4, interferon-gamma (IFN gamma) and/or SCH 42427 (SCH), a new triazole antifungal agent, and their effect on serum IgE levels in a murine model of chronic Paracoccidioides brasiliensis infection. BALB/c mice infected by the pulmonary route were studied with three programmes. The subacute model and one acute model experiment investigated cytokine secretion by lymph node cells (LNC), and in a second acute experiment mice were given anti-IL-4, IFN-gamma or nothing 24 h post infection, then killed at 4 weeks. In the chronic model, mice began treatment at 4 weeks post infection, receiving either SCH, IFN-gamma alone, SCH+IFN-gamma, or no treatment for 8 weeks. At 2-week intervals lung and spleen burdens of infection and serum polyclonal IgE levels were determined. In the subacute model (non-progressive infection), initially there was dual production of IL-4 and IFN-gamma by antigen stimulated LNC. In the acute progressive infection model IL-4, but not IFN-gamma, was secreted. Anti-IL-4 treatment of the acute phase resulted in enhanced host resistance to infection, which correlated with decreased serum IgE. The chronic model, in which the in vivo efficacy of SCH against P. brasiliensis was shown, suggests possible synergy between immunomodulation and antimicrobial chemotherapy (IFN-gamma and SCH). Decreased organ burdens of infection in the chronic model after treatment with SCH, SCH plus IFN-gamma, or anti-IL-4 correlated with decreased serum IgE. These promising novel approaches to treatment of systemic fungal infections suggest a Th2 type of response to P. brasiliensis infection, which can be reversed with successful therapy. PMID- 8403492 TI - The functional affinities of antibodies of different IgG subclasses to dietary antigens in mothers and their babies. AB - The quantity and functional affinities of IgG1 and IgG4 antibodies to the dietary antigens casein and ovalbumin were measured in unselected mothers and their 1 year-old infants. In these infants, the titre of IgG antibodies to both antigens was highest in the IgG1 subclass, while in their mothers the titre of IgG1 and IgG4 antibodies to these foods was similar. High affinity IgG4 responses to both casein and ovalbumin were frequently found in mothers, whilst IgG1 responses, particularly to ovalbumin, were of low functional affinity. By contrast, in the 1 year-old infants, the functional affinity of IgG1 antibody to ovalbumin was substantially higher than in their mothers (Student's paired t-test, P < 0.001), indicating that higher affinity IgG1 antibody was produced on first exposure to ovalbumin rather than following chronic exposure. The effect of antigenic load on affinity maturation was further investigated by comparing the affinity of IgG1 antibody to casein in bottle, mixed and breast fed infants. Bottle fed infants had significantly higher-affinity IgG1 antibodies to casein compared with breast or mixed fed infants (Student's unpaired t-test, P < 0.01 and 0.02), suggesting that antigen exposure via the gut was able to drive the affinity maturation process. In studying the immune response it is clear that account must be taken of the affinity as well as of the titre of the antibody produced. PMID- 8403493 TI - Analysis of cytokine profiles in synovial T cell clones from chlamydial reactive arthritis patients: predominance of the Th1 subset. AB - Subpopulations of human T cells (Th0, Th1 and Th2) can be distinguished by their cytokine-secretion pattern. Evidence is increasing from other studies that the outcome of a human disease may depend on the subpopulation of T cells that predominates at the site of inflammation. Reactive arthritis serves as a useful model of chronic inflammatory diseases, because the triggering antigen can be identified. Using this triggering antigen we raised 33 T cell clones reactive with Chlamydia trachomatis and 25 T cell clones that were not reactive, all from the synovial fluid of two patients suffering from Chlamydia-induced arthritis. Their cytokine secretion patterns for interferon-gamma (IFN-gamma), IL-2 and IL-4 were analysed, as also were mRNAs for IFN-gamma and IL-10 by in situ hybridization. Out of the 33 antigen-reactive clones 23 showed a Th1 pattern with IFN-gamma but not IL-4 secretion, while the remaining 10 exhibited a Th0 pattern. The clones that did not react with Chlamydia expressed all patterns of cytokine secretion, including a Th2 pattern, thus providing a control population that excludes bias in the sampling procedure. CD4 and CD8 clones displayed a similar cytokine-secretion pattern. In addition this study demonstrates for the first time the expression of IL-10 mRNA in T cell clones derived from synovial fluid, and this was not confined to the Th2 subset. The Th1 response that Chlamydia provoke can be regarded as appropriate for such an obligate intracellular pathogen. PMID- 8403494 TI - Bronchoalveolar lavage cell analysis and lung function impairment in patients with systemic lupus erythematosus (SLE). AB - We examined the relationship between peripheral blood and bronchoalveolar lavage (BAL) lymphocyte phenotypes and lung function in 19 patients with SLE, and evaluated their association with disease activity. Lung function assessment showed a mildly restrictive pattern with frequent impairment of transfer factor for carbon monoxide (T1,co) and diffusing capacity of the alveolocapillary membrane (Dm), of late-expiratory airflow rates and with a high prevalence of increased airway resistance. T1,co, Kco and Dm correlated inversely with the numbers of CD8+ cells and CD56+/CD16+/CD3- (NK) cells in BAL. Oxygen radical production, both by stimulated and unstimulated BAL cells and blood polymorphonuclear leucocytes (PMN) was significantly increased in SLE. In comparison with healthy controls, patients with SLE had a lower percentage of CD19+ B cells in the BAL versus an increased percentage of these cells in peripheral blood. HLA-DR expression on CD4+ and CD8+ lung lymphocytes was markedly increased in SLE. Current SLE disease activity was not associated with changes in BAL or peripheral blood lymphocyte phenotypes. Our data suggest that an ongoing cell-mediated immune response is present in the lungs in SLE, particularly involving activated CD8+ T cells and CD56+/CD16+/CD3- NK cells. It is associated with up-regulated local production of oxygen radicals and with impaired pulmonary diffusing capacity. This inflammatory process seems to be independent of general SLE disease activity. PMID- 8403495 TI - Polynucleotide specificity of anti-reactive oxygen species (ROS) DNA antibodies. AB - Hydrogen peroxide in the presence of short wavelength UV light was able to induce alterations in native DNA fragments of 300 bp (ROS-DNA), thereby rendering it immunogenic in experimental animals. The specificity of induced antibodies was investigated by direct binding and competition ELISA. Inhibition studies revealed nearly 89% inhibition in the antibody binding by the immunogen and recognition of native B-, A- and allied conformations presented by various synthetic polynucleotides. Gel retardation assay reiterated the formation of immune complexes between induced antibodies and native and ROS-DNA fragments. It was observed that naturally occurring anti-DNA autoantibodies from systemic lupus erythematosus (SLE) sera recognize ROS-DNA. The comparison of the specificities of anti-DNA autoantibodies from 10 SLE patients showed a 20-50-fold preference for ROS-DNA over native DNA. These results demonstrate that anti-DNA antibodies can be induced by ROS-DNA, and that some of the autoimmune DNA binding antibodies found in SLE may result from response to reactive oxygen species. PMID- 8403496 TI - Triggering of respiratory burst by phagocytosis in monocytes of patients with systemic lupus erythematosus. AB - The triggering of the respiratory burst by phagocytosis via different receptors in monocytes of patients with systemic lupus erythematosus (SLE) was investigated. The superoxide anion synthesis was assayed by reduction of ferricytochrome C that was inhibited by superoxide dismutase. The mononuclear cell suspensions were triggered by IgG-coated latex, C3 complement fragment coated and uncoated yeast (Saccharomyces cerevisiae). Superoxide generation induced by phagocytosis via Fc gamma R was decreased in monocytes of patients with SLE. On the other hand, MoAbs against Fc gamma RI, Fc gamma RII and especially CR3 could also induce superoxide anion synthesis. At the same time, superoxide generation induced by anti-CR3 could be inhibited with C3-coated yeast. PMID- 8403497 TI - The effects of cytokines on metalloproteinase inhibitors (TIMP) and collagenase production by human chondrocytes and TIMP production by synovial cells and endothelial cells. AB - It has been suggested that IL-1 produces cartilage matrix degradation by metalloproteinases such as collagenase, and that such degradation is regulated by metalloproteinase inhibitors (TIMP). Therefore, the balance between collagenase and TIMP is an important factor for tissue destruction in inflammatory joints. In the present study the effects of cytokines on collagenase and TIMP production in chondrocytes as well as the effects of cytokines on TIMP production in connective tissue cells were studied. IL-1 beta inhibited TIMP production in endothelial cells while enhancing TIMP production in synovial cells and chondrocytes. In addition, tumour necrosis factor-alpha (TNF-alpha) significantly inhibited and IL 6 significantly enhanced TIMP production in endothelial cells, synovial cells and chondrocytes. In the chondrocyte supernatant, collagenase activity/TIMP ratio was significantly elevated by the addition of either IL-1 beta or TNF-alpha to the cells, whereas the ratio was significantly decreased by IL-6. These results suggest that the cytokine effects on TIMP production are different among the different cell types, and that either IL-1 beta or TNF-alpha induce cartilage matrix degradation by disrupting the collagenase/TIMP balance, while, on the other hand, IL-6 protects the tissue through an opposite effect. PMID- 8403498 TI - Regulation of resistance against adjuvant arthritis in the Fisher rat. AB - Inbred female Lewis (LEW/N) rats develop a severe chronic arthritis in the adjuvant arthritis (AA) model, histocompatible Fisher (F344/N) rats are resistant and germ-free Fishers (GF F344) are again susceptible. In this study we show that the F344 rat can become susceptible to AA, using Mycobacterium tuberculosis (M.tb.) in the powerful adjuvant paraffin oil, instead of mineral oil (Freund's incomplete adjuvant (FIA)). This indicates that the F344 rat does not lack T effector cells. To examine further mechanisms responsible for suppression, we determined the level of plasma corticosterone in response to IL-1 alpha in Lewis, F344 and GF F344 rats. IL-1 alpha induced only low amounts of corticosterone in Lewis rats, but high amounts in both F344 and GF F344 rats. The GF F344 rats are susceptible to AA, but the severity of the disease is reduced compared with Lewis rats. This indicates that corticosterone may be an important mechanism to suppress disease development, but not the only mechanism. In addition we investigated whether T suppressor cells play a role in the resistance of the F344 strain. This was performed by pretreating the animals with the immunomodulating drugs cyclophosphamide (Cy) and cyclosporin A (CsA). We were unable to make the F344 rat susceptible to AA, indicating that active suppression does not play a role in the induction phase of arthritis. This finding is confirmed in adoptive transfer experiments of AA from Lewis to F344 rats. Our data suggest the lack of a strong pre-existing suppression in the F344 rats, and indicate that suppression is generated upon bacterial challenge. Whether suppression is overruled probably depends on the power of adjuvants used and potential control by corticosteroids. PMID- 8403499 TI - Modulation of alpha 1-acid glycoprotein (AGP) gene induction following honey bee venom administration to adjuvant arthritic (AA) rats; possible role of AGP on AA development. AB - Honey bee venom (HBV) administration to adjuvant arthritic (AA) rats resulted in a significant suppression of arthritis and in suppression of the hepatic acute phase alpha 1-acid glycoprotein (AGP) gene induction at the early stages of disease development. AGP administration in AA rats resulted in acceleration of arthritis development and in increase of severity and duration of the disease. IL 1, IL-6, tumour necrosis factor (TNF) and glucocorticoids alone are not responsible for the HBV-mediated AGP gene down-regulation. These results indicate that AGP gene expression in AA and HBV-treated AA rats involves the interaction of several factors, and that AGP plays a role for AA development in rats. PMID- 8403500 TI - Remission of experimental autoimmune hepatitis is associated with antigen specific and non-specific immunosuppression. AB - Experimental autoimmune hepatitis (EAH) is an animal model for autoimmune hepatitis. The disease is T cell-mediated and runs a subacute course, with maximal disease activity around week four after disease induction and a slow ensuing recovery. The aim of the present study was to investigate the immunoregulatory mechanisms that may account for recovery in EAH. It was found that T cell reactivity to liver antigens preceded histological disease, but at the peak of disease activity this T cell response was already suppressed. Active and antigen-specific suppression could be demonstrated, as irradiated splenocytes from animals at the beginning of recovery from EAH were able to suppress in vitro the T cell response to liver antigens by 42%. The response to an irrelevant antigen was suppressed by 16%, showing additional antigen non-specific suppression in vitro. The response to an in vivo immunization with an unrelated microbial antigen (tetanus toxoid) at the peak of disease was markedly reduced (by 90%). These data demonstrate in vitro and in vivo immunosuppression associated with the overcoming of and recovery from EAH. They stress the importance of immunoregulatory cycles in the control of autoimmune hepatitis. PMID- 8403501 TI - Analysis of the in vitro cytokine production by liver-infiltrating T cells of patients with autoimmune hepatitis. AB - The pathogenic mechanisms underlying the development of autoimmune hepatitis (AIH) are still unclear. Since AIH is associated with the presence of various autoantibodies and certain HLA subtypes, it is likely that T and B cells play a major role in this disease. In this study we have determined the functional capacities of in vivo preactivated liver-infiltrating T cells (LTC) from patients with AIH. As controls we used LTC from patients with non-autoimmune hepatitis (non-AIH). Our results show that preactivated LTC from patients with AIH predominantly (190/255 clones) reside in the CD4+ population, whereas LTC in non AIH are dominated by the CD8+ phenotype (148/254 clones). In view of this finding we have investigated the cytokine secretion patterns of 102 randomly chosen CD4+ T cell clones from six patients with AIH. As controls we have used 58 CD4+ LTC from 11 patients with non-AIH. All clones were stimulated by lectin and irradiated accessory cells and subsequent cytokine production was evaluated. LTC from patients with AIH have a lower interferon-gamma (IFN-gamma)/IL-4 ratio compared with LTC from non-AIH. Although clones from some patients with AIH produced very high amounts of IL-4 in vitro, this was not a constant finding. These results show that in vivo preactivated LTC from patients with AIH are mostly CD4+ T cells that produce more IL-4 than IFN-gamma. In contrast, LTC from patients with non-AIH are dominated by CD8+ and CD4+ T cells that produce significantly less IL-4 than IFN-gamma. Thus, liver-infiltrating T cells from patients with AIH and non-AIH belong to different functional T cell subsets. This may have implications for the regulation of humoral and cellular immune responses in inflammatory liver disease. PMID- 8403502 TI - HIV induces deletion of T cell receptor variable gene product-specific T cells. AB - Using a flow cytometric method, CD4+, CD8+, alpha beta TCR+ and TCR variable region gene product (TVRGP)-specific T cells were analysed in healthy heterosexual males (HHeM), HIV-seronegative homosexual males (SNHM), asymptomatic seropositive homosexual males (ASPH) and homosexual males with AIDS who were either well (AIDS-A), or unwell in hospital (AIDS-B). Total CD4+ and CD8+ T cell numbers were similar in HHeM and SNHM. CD4+ T cells were significantly reduced in ASPH relative to both HHeM and SNHM and in AIDS-A and AIDS-B relative to SNHM. TVRGP-specific T cells expressed as a percentage of TCR alpha beta + cells showed no significant difference in HHeM, SNHM and AIDS-B. The proportion of alpha beta + cells expressing the V beta 5.1, V beta 12 and V alpha 2 gene product (GP) was, however, significantly reduced in ASPH and AIDS-B relative to HHeM, SNHM and AIDS A. Possible causes of TVRGP-specific T cell deletion are discussed. PMID- 8403503 TI - Enhanced secretion of tumour necrosis factor-alpha, IL-6, and IL-1 beta by isolated lamina propria mononuclear cells from patients with ulcerative colitis and Crohn's disease. AB - The perpetuation of inflammation in ulcerative colitis and Crohn's disease may be regulated in part by an increased secretion of proinflammatory cytokines due to either an appropriate response to initial stimulating agents, and/or due to an impaired down-regulation of cytokine secretion. The aim of this study was to determine the secretion patterns of the proinflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), IL-6 and IL-1 beta, from isolated lamina propria mononuclear cells (LPMNC) isolated from colonic biopsies from patients with untreated ulcerative colitis or Crohn's disease. LPMNC isolated from involved inflammatory bowel disease (IBD) mucosa spontaneously produced increased amounts of TNF-alpha, and IL-6, and IL-1 beta. The TNF-alpha secretion from IBD LPMNC could be further enhanced by pokeweed mitogen stimulation. The secretion patterns of TNF-alpha and IL-1 beta by LPMNC from patients with either ulcerative colitis or Crohn's disease demonstrated a close correlation with the degree of tissue involvement and mucosal inflammation. LPMNC from non-involved ulcerative colitis mucosa secreted markedly increased levels of IL-6 compared with non-involved Crohn's disease mucosa or control mucosa. The heightened IL-6 secretion from LPMNC from non-involved ulcerative colitis mucosa without visible or microscopic signs of inflammation indicates that the pathophysiologic mechanisms involved in the initiation of inflammation may differ between ulcerative colitis and Crohn's disease. The determination of proinflammatory cytokine secretion by isolated LPMNC from colonoscopic biopsies may be a sensitive method for monitoring the severity of mucosal inflammation in IBD patients. PMID- 8403504 TI - Immunohistochemical demonstration of membrane cofactor protein (MCP) of complement in normal and diseased kidney tissues. AB - The immunohistochemically stained membrane cofactor protein of complement (MCP/CD46), one of the complement regulatory proteins, was up-regulated in some diseased kidney tissues. MCP in diseased kidneys was strongly concentrated along the glomerular capillary walls as well as in the mesangial regions, while MCP in normal kidneys was weakly detected in all glomerular structural cells and in the epithelial cells of tubules. Since the enhanced staining was noted in those areas where depositions of C3b/C3c occurred, ongoing complement reaction might be responsible for the up-regulation of MCP expression. MCP expression may be up regulated by complement fragments generated during complement activation in glomerulonephritis. Furthermore, anti-MCP staining was stronger in intensity in patients with moderate to massive proteinuria, indicating that up-regulation of MCP expression could be directly correlated to the kidney damage. PMID- 8403505 TI - On the association between amyloid fibrils and glycosaminoglycans; possible interactive role of Ca2+ and amyloid P-component. AB - We have previously reported the specific association of glycosaminoglycans (GAG) and proteoglycans (PG) with amyloid fibrils and characterized the polysaccharides directly extracted from amyloid-laden tissues. In the present study we further elucidate the association between purified amyloid fibrils and GAG/PG with special reference to those GAG/PG associated with amyloid P-component (AP) and the interactive role of Ca2+ ions. Amyloid fibrils were isolated from human hepatic AA amyloid employing water extraction with and without preceding removal of AP, an extrafibrillar protein component of all amyloids, using sodium citrate. GAG/PG co-isolated with the amyloid extracts, with and without AP, were isolated and characterized. Agarose-affinity chromatography of extracts containing AP was performed, and the GAG associated with this extrafibrillary protein were characterized as well. Several different GAG/PG populations were demonstrated in the various extracts. The abolition of calcium-dependent binding markedly influenced the amount of GAG/PG recovered in the fibril extracts, as well as the total amount of amyloid material obtained. Thus, it seems that calcium plays an important role in the association between the fibrils and the sugar moieties, and that a significant fraction of the GAG found in amyloid exhibits a Ca(2+) dependent fibril-GAG interaction. No significant difference in the proportion between galactosaminoglycans and glucosamines was, however, disclosed when the two extraction protocols were compared, suggesting that no particular GAG species has a higher affinity for the fibrils themselves. Both dermatan/chondroitin sulphate and heparan sulphate identified in the present study exhibited a Ca(2+) dependent interaction with AP, supporting previous findings. However, the amyloid associated galactosaminoglycans found, especially the large PG appearing in small amounts, seemed to have a higher affinity for the extrafibrillar AP than the other GAG. PMID- 8403506 TI - Measurement of antiphospholipid antibody by ELISA using purified beta 2 glycoprotein I in preeclampsia. AB - It has been reported that the binding of some antiphospholipid antibodies (APA) to phospholipids requires the presence of beta 2-glycoprotein I (beta 2-GPI). Using a new ELISA, in which well coated phospholipids were treated with a constant amount of purified beta 2-GPI, we tried to detect the presence of APA which binds to phospholipid/beta 2-GPI complex or to phospholipids such as cardiolipin (CA) and phosphatidylserine (PS) in preeclampsia, and to check for clinical abnormalities in antibody-positive cases. Serum samples were taken from 43 cases of preeclampsia, including 26 cases of the severe type, and 47 normal pregnant women. Positive rates of anticardiolipin antibody (ACA) by ELISA using CA/beta 2-GPI complex in mild, severe and total preeclampsia were 20.0, 17.4% and 18.4% respectively. No antibody-positive cases were found in normal pregnancies. ACA was detected much more frequently when cardiolipin/beta 2-GPI complex was used in ELISA compared with ELISA without beta 2-GPI. Positive rates of antiphosphatidylserine antibody (APSA) in mild, severe and total preeclampsia were 5.9%, 11.5% and 9.3% respectively. APSA was also detected much more frequently when the phosphatidylserine/beta 2-GPI complex was used in ELISA. The frequency of intrauterine growth retardation (IUGR) in the ACA-positive subjects was higher than that of ACA negatives. We suggest that the ACA and APSA which bind to phospholipid/beta 2-GPI complex are detectable in preeclampsia, and that these antiphospholipid antibodies are related to fetal growth. PMID- 8403507 TI - Population study of T cell receptor V beta gene usage in peripheral blood lymphocytes: differences in ethnic groups. AB - The T cell receptor (TCR) V beta repertoire in peripheral blood lymphocytes (PBL) of a large number of healthy individuals was analysed by quantifying V beta specific mRNA using the method of anchored multiprimer DNA amplification and a reverse dot blot assay. Among 16 V beta gene families examined, particular V beta genes were noted to be unequally expressed in the PBL of 70 healthy donors. The frequently used genes belong to the V beta 4, 5, 6, 8 and 13 (12) families, while V beta 1, 9 and 15 were the least frequently used gene families. This bias in gene usage was observed in all individuals. Marked deviation from the mean percentage usage was noted for some V beta genes in individuals when their PBL were examined serially, but the common pattern of biased usage was not grossly distorted. When the TCR repertoire of different ethnic groups was examined, a lower mean frequency of V beta 3.2 was seen in the repertoire of 19 Caucasians compared with 25 age-matched Samoans (P < 0.003). Conversely, the expression of V beta 5.1 and V beta 5.3 was higher in Caucasians than in 51 age-matched Polynesians (Maoris and Samoans, P < 0.003). Considering the 20% co-efficient of variation in the estimate of V beta gene usage, our data from 70 unrelated individuals suggest that in PBL, individual variations in the TCR repertoire were superimposed upon a common biased usage of V beta genes in the general population. PMID- 8403508 TI - Effect of low-dose ultraviolet-B radiation on the function of human T lymphocytes in vitro. AB - Purified peripheral blood human T lymphocytes, derived from normal individuals, were assayed for their susceptibility to low doses of ultraviolet B (UVB) in vitro. Exposure of T cells to graded single doses (range 0-8 mJ/cm2) of UVB resulted in a dose-dependent reduction of viability. This phototoxic effect was not immediately apparent, however, but became manifest 48-72 h subsequent to irradiation. A dose as little as 0.5-1 mJ/cm2 was sufficient to cause 50% mortality. Irradiated T cells showed a reduced ability to proliferate, irrespective of the stimulus used, and a reduced ability to produce cytokines IL 2, IL-4, IL-5, interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF alpha). This decreased ability was UVB-dose related and, remarkably, was exactly correlated to phototoxicity. UVB had no effect on CD4 and CD8 expression or their ratio, whereas the expression of IL-2R (CD25) was only slightly reduced. Our data suggest that UVB radiation neither selectively affects Th1 or Th2 nor CD4 or CD8 T cell subsets. The high susceptibility of T cells to UVB might explain, at least in part, the beneficial effect of phototherapy during treatment of certain immunodermatological diseases. PMID- 8403509 TI - In vitro cytotoxicity as a marker of hypersensitivity to sulphamethoxazole in patients with HIV. AB - Hypersensitivity to trimethoprim-sulphamethoxazole (TMP-SMX) in patients with HIV infection may be a result of either immune dysregulation, a direct cytotoxicity of the SMX-hydroxylamine metabolite (SMX-HA) (rather than SMX per se), or glutathione deficiency. We evaluated the in vitro cytotoxicity of SMX and SMX-HA to peripheral blood mononuclear cells (PBMC) of HIV-infected subjects to determine if the degree of in vitro cytotoxicity is associated with hypersensitivity, whether glutathione inhibits cytotoxicity, and whether in vitro cytotoxicity is predictive for the development of hypersensitivity. Given that fever is often a prominent feature of hypersensitivity, we also assessed whether SMX or SMX-HA could induce the in vitro production of IL-1 beta, IL-6 or tumour necrosis factor-alpha (TNF-alpha) by PBMC. The cytotoxicities of SMX and SMX-HA to PBMC were assessed in 45 HIV-infected patients with prior TMP-SMX therapy, and in eight HIV- controls. Twelve HIV-infected subjects were studied prospectively before primary Pneumocystis carinii pneumonia (PCP) therapy or rechallenge with TMP-SMX in previously hypersensitive subjects. Cytokine production was measured in four hypersensitive and two non-hypersensitive HIV-infected subjects, and three HIV-uninfected controls. The cytotoxicity of SMX-HA to PBMC was significantly greater in the 22 HIV-infected patients with prior hypersensitivity than both the 23 HIV-infected patients without hypersensitivity and the control group. Cytotoxicity was significantly reduced by glutathione only in the hypersensitive group. SMX did not induce cytotoxicity in any group. In 12 subjects studied prospectively, SMX-HA cytotoxicity was also significantly greater in those with subsequent hypersensitivity. Exposure of PBMC to SMX-HA resulted in a modest increase in the production of IL-6, IL-1 beta and TNF-alpha, although no major difference was detected between subjects with or without hypersensitivity. These data suggest that SMX-HA and glutathione deficiency are involved in the pathogenesis of hypersensitivity to TMP-SMX in HIV-infected patients, and that in vitro cytotoxicity could be useful in the diagnosis of hypersensitivity and predicting its likelihood. PMID- 8403510 TI - Effects of activin A on IgE synthesis and cytokine production by human peripheral mononuclear cells. AB - Activin A not only stimulates the synthesis and release of pituitary follicle stimulating hormone, but exerts various effects on haematopoietic cells, embryos, and fibroblasts. In the present study we have examined effects of activin A on IgE synthesis and cytokine production by peripheral blood mononuclear cells (PBMC) in normal humans. When PBMC were cultured in the presence of IL-4, activin A significantly augmented IgE production induced by IL-4. Activin A did not affect, however, IgE production from highly purified B cells when they were stimulated with anti-CD40 MoAb and IL-4. The fact that in the latter condition IgE synthesis was T cell- and monocyte-independent indicated that activin A does not directly influence B cells for IgE synthesis. Rather, production as well as gene expression of IL-6, which is known to enhance IgE synthesis by purified monocytes, was induced by activin A alone. In addition, activin A induced other monokines such as IL-1 and tumour necrosis factor (TNF)-alpha from monocytes. In contrast, activin A neither induced nor augmented the production of TNF-beta or interferon-gamma (IFN-gamma), both of which are known to be exclusively generated by T cells. These data indicate that activin A plays a certain role in physiological functions for monocytes in normal humans. PMID- 8403511 TI - Transforming growth factor-beta (TGF-beta) in human milk. AB - The amount of TGF-beta contained in human whey was studied by the colony formation of NRK47F cells. It was noted that a factor inducing colony formation did exist in human whey, and its action was neutralized when anti-TGF-beta antibodies were introduced. This suggests that TGF-beta does exist in human whey. In colostrum, the total amount of TGF-beta was 1365.7 +/- 242.9 ng/ml, of which the active form comprised 728.1 +/- 248.7 ng/ml (n = 21). In late milk, the total TGF-beta was 952.5 +/- 212.6 ng/ml, with an active form of 178.7 +/- 157.3 ng/ml. Thus human milk contains a large amount of active TGF-beta. Furthermore, it was revealed by the reverse transcriptase polymerase chain reaction that mRNAs coding TGF-beta 1 and TGF-beta 2 exist in human milk cells. These results suggest that both TGF-beta 1 and TGF-beta 2 exist in human milk. PMID- 8403512 TI - Blockade of immunoregulatory Fc-signalling by HIV peptides: oligopeptides from HIV gp120 and gp41 bind the Fc portion of IgG and increase the in vitro anti ssDNA response. AB - Concomitant ligation of antigen receptors with Fc-receptors negatively signals B cells. Antibodies to the Fc portion of IgG prevent this negative Fc-signalling, provided that these antibodies do not emit Fc signals. Prevention of Fc signals leads to augmented antibody responses to self and foreign antigens, and reduces the requirement for T cells by 10- to 100-fold in T cell-dependent antibody responses. In ELISA assays, peptides from conserved portions of the glycoproteins, HIV-1 gp120 or gp41 from HIV-1 and HIV-2 bind to the Fc portion of IgG, but do not bind the F(ab')2 portion of IgG. HIV-derived peptides, which bind to the Fc portion of IgG, augment the antibody-forming cell response to single stranded (ss)DNA. The spontaneous response to ssDNA using spleen cells from young mice, and the response in the presence of exogenous DNA using spleen cells from old mice, are augmented to the greatest extent. These results demonstrate that HIV peptides bind to the Fc portion of IgG and augment immune responses to DNA; they suggest the possibility that blockade of the Fc portion of IgG antibodies is associated with a reduction in Fc-mediated regulation of anti-self responses. Blockade of regulatory Fc-signalling may account for increased circulating immunoglobulins and autoantibodies in clinical AIDS. PMID- 8403513 TI - Dendritic cells infected in vitro with human T cell leukaemia/lymphoma virus type 1 (HTLV-1); enhanced lymphocytic proliferation and tropical spastic paraparesis. AB - Evidence supporting a role of the dendritic cell (DC) in stimulating autologous T cell activity in tropical spastic paraparesis (TSP) was sought by studies of cells taken from healthy volunteers and exposed to HTLV-1 in vitro. DC were co cultured with an HTLV-1-producing cell line (MT-2) at 1:1 or 10:1 ratios. These DC stimulated high levels of proliferation in autologous T cells. This was similar to that seen in an autologous mixed leucocyte reaction (AMLR) using cells from TSP patients. The requirement for both DC and virus was confirmed, since neither DC co-cultured with uninfected MT-2 cells nor addition of infected MT-2 cells directly to T cells caused significant stimulation. DC exposed to the highest dose of HTLV-1 (1:1) for 24 h before addition of T cells caused strong stimulation that increased after 8 h but almost disappeared by 72 h. In situ hybridization showed that approximately 25% of DC became infected in cultured cells after preincubation for 24 h, and over 50% were infected with a 72-h preincubation. We suggest that infection of DC by HTLV-1 may be an initial step in altering the immune system in seronegative patients, and that persistent T cell stimulation in those with genetic susceptibility may underlie the production of neurological disease. PMID- 8403514 TI - Selective impairment of T lymphocyte activation through the T cell receptor/CD3 complex after cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is reported to cause transient immunosuppression in man. In this study we have analysed the effect of CMV on T lymphocyte function in 29 children diagnosed for acute CMV infection. Peripheral blood mononuclear cells (PBMC) obtained from the patients showed a significant specific impairment in their proliferative response to enterotoxins A and C1, to concanavalin A and to the anti-CD3 MoAb OKT3. The impaired responses were corrected with exogenous IL-2. In contrast, stimulation using phytohaemagglutinin, as well as activation signals delivered through the surface molecules CD26 or CD28, elicited normal proliferative responses in CMV PBMC. The results indicate that the T cell anergy associated with CMV infection is restricted to the T cell receptor/CD3 activation pathway. PMID- 8403515 TI - Peri-operative modulation of cellular immunity in patients with colorectal cancer. AB - The peri-operative cellular immune response is depressed in patients with gastrointestinal cancer, a factor which may facilitate malignant dissemination. We have investigated the effects of peri-operative rIL-2 and a combination of rIL 2 and interferon-alpha (IFN-alpha) on both peripheral blood lymphocyte function and number in patients undergoing surgical resection for colorectal cancer. Fifty two patients were randomly allocated to either control, rIL-2 or rIL-2 with IFN alpha treatment arms. In vitro studies were performed pre-operatively and on post operative days 1, 4, 7 and 10. Natural killer (NK) and lymphokine-activated killer (LAK) cell function were profoundly depressed in control patients (P < 0.001; P < 0.01), an effect abrogated in both treatment groups; indeed NK function was augmented in the rIL-2 and IFN-alpha group on the first post operative day in association with an increase in the percentage of cells expressing CD16 and CD56 (P < 0.01). Flow cytometric analysis of lymphocyte subsets in the control group was unremarkable, except for an early post-operative fall in numbers of lymphocytes. Treatment with either rIL-2 or rIL-2 and IFN alpha produced an initial profound reduction in T lymphocyte numbers, followed by a 'rebound' lymphocytosis of activated CD3+ T cells, as demonstrated by a significant increase in co-expression of CD25, CD38 and CD45RO. No significant differences were observed between either of the treatment groups. Adjuvant immunotherapy affects peri-operative anti-tumour immune responses, and this may influence long term outcome in patients undergoing surgery for gastrointestinal cancer. PMID- 8403516 TI - Up-regulation of alveolar macrophage platelet-derived growth factor-B (PDGF-B) mRNA by interferon-gamma from Mycobacterium tuberculosis antigen (PPD)-stimulated lymphocytes. AB - Macrophage production of PDGF-B is believed to be important in the pathogenesis of diseases where chronic lung inflammation develops into fibrosis. Since tuberculosis is characterized by chronic inflammation and tissue fibrosis, we asked if lymphokines from lymphocytes stimulated by the Mycobacterium tuberculosis antigen PPD, contained factors capable of increasing human alveolar macrophage PDGF-B mRNA. Supernatants from both phytohaemagglutinin (PHA)- and purified protein derivative (PPD)-stimulated lymphocytes, when added to macrophages, induced an increase in the mRNA of PDGF-B, but not transforming growth factor-beta (TGF-beta). When lymphocytes from contacts of patients with tuberculosis, patients with tuberculosis, and normal subjects were compared following PPD stimulation, the lymphocytes from the contacts had the greatest proliferation response, the greatest production of interferon-gamma (IFN-gamma), and their lymphokines induced the greatest increase in PDGF-B mRNA in macrophages. Recombinant human IFN-gamma reproduced this ability of lymphokines to increase macrophage PDGF-B mRNA. Finally, the increase in macrophage PDGF-B mRNA following incubation with supernatants from PPD-stimulated lymphocytes was shown to be due to IFN-gamma, when the increase in macrophage PDGF-B mRNA was prevented by addition of anti-human IFN-gamma antibody to the lymphocyte supernatant. This study indicated that antigen-stimulated lymphocytes released IFN-gamma, which in turn resulted in an increase in PDGF-B mRNA in alveolar macrophages. Such a mechanism provides a link between the DTH response and the first stages of a fibrotic reaction, and may offer an explanation for the progression of chronic inflammation to fibrosis, as occurs in the lungs of patients with untreated pulmonary tuberculosis. PMID- 8403517 TI - Increased expression and occupancy of receptors for tumour necrosis factor on blood monocytes from tuberculosis patients. AB - Blood monocytes from tuberculosis patients release high amounts of tumour necrosis factor-alpha (TNF-alpha). Because the biological efficiency of TNF-alpha would depend on the expression of TNF-alpha receptors on target cells, we thought to analyse the capacity of blood monocytes from a group of patients with pulmonary tuberculosis to bind 125I-TNF-alpha. We report a slight but not significant enhancement in specific binding of 125I-TNF-alpha on monocytes of 15 consecutively studied patients compared with 10 controls. Per cent cell surface bound and internalized 125I-TNF-alpha was identical in the two groups. To evaluate the receptor occupancy by endogenously generated TNF-alpha, similar experiments were performed after cell exposure to low-pH glycine buffer. Under these conditions, specific binding of 125I-TNF-alpha was significantly higher on tuberculosis monocytes compared with control monocytes. Moreover, the occupancy of TNF-alpha receptors by endogenously generated TNF-alpha that was found to be significantly higher on tuberculosis monocytes than on control monocytes, was directly related to the enhanced capacity of mononuclear cells to generate TNF alpha in vitro. It normalized after 3 months of antituberculous therapy. Scatchard analysis of the binding data revealed that tuberculosis infection caused a significant increase in high affinity 125I-TNF-alpha binding to monocytes without any significant change in the dissociation constant. Collectively, these results indicate an up-regulation of TNF-alpha generation and binding to blood monocytes in patients with pulmonary tuberculosis. They provide support to the hypothesis that TNF-alpha is of critical importance in the pathogenesis of this infection. PMID- 8403518 TI - Effect of lipoarabinomannan and mycobacteria on tumour necrosis factor production by different populations of murine macrophages. AB - Tumour necrosis factor (TNF) production is an important pathological mediator in mycobacterial infections, and yet little is known of the factors which influence its production. We have studied the influence of murine macrophage heterogeneity and activation state on TNF production following mycobacterial stimulation in vitro. Lipoarabinomannan (LAM) from strains of Mycobacterium tuberculosis and Myco. avium differentially stimulated TNF production in thioglycollate-elicited macrophages in a dose-dependent manner. In comparison, resident peritoneal macrophages produced much less TNF when stimulated with LAM, dead mycobacteria or lipopolysaccharide (LPS). In contrast, zymosan stimulated resident macrophages to a higher degree than thioglycollate-elicited cells. Another comparison between bone marrow and thioglycollate-elicited macrophages showed that both responded to LPS, but only the latter was stimulated significantly by H37Rv LAM. This may indicate that LAM stimulation of macrophages takes place through a different pathway than both zymosan- and LPS-stimulated TNF production. Also, in vitro activation of peritoneal macrophages with interferon-gamma (IFN-gamma), increased TNF response to several stimuli. Our studies indicate that the pathology of mycobacterial infections through TNF production may be influenced by the type and activation state of the macrophage which responds to that infection. PMID- 8403519 TI - Lymphoproliferative responses to a merozoite surface antigen of Plasmodium falciparum: preliminary evidence for seasonal activation of CD8+/HLA-DQ restricted suppressor cells. AB - We have investigated the phenotype of human lymphocytes responding to a defined Plasmodium falciparum malaria antigen in vitro. Cells were obtained from the peripheral blood of malaria-immune donors from an endemic area of West Africa and were tested for proliferation in response to cloned fragments of a merozoite surface protein (PfMSP1). Depletion and inhibition studies indicated that the majority of proliferating cells were CD4+ and restricted by HLA-DR or -DQ. A proportion of responding cells appeared to be CD8+, but their response was dependent on help from CD4+ cells. In two donors there was evidence that low responses could be enhanced by removal of CD8+ cells and/or blocking of antigen presentation by anti-HLA-DQ antibodies. This phenomenon was observed in cells collected during the wet (malaria transmission) season but not in cells collected from the same individual during the dry season. PMID- 8403520 TI - Seroreactivity with the Plasmodium falciparum blood stage antigen Pf332 in adults and children from malaria-endemic regions. AB - It has earlier been reported that the human monoclonal antibody (MoAb 33G2) and polyclonal antibodies reactive with Pf332 may interfere in vitro with the erythrocytic cycle of Plasmodium falciparum at two potential target sites for protective antibodies, indicating that the antigen may constitute an important target for immune responses during malaria infections. MoAb 33G2 shows its highest reactivity with repeated sequences in the antigen Pf332 and also cross reacts with determinants in Pf155/RESA. This study was conducted in order to assess the prevalence of seroreactivity against Pf332 in individuals residing in areas of different malaria endemicity, and in children with different degrees of disease severity. We now report that individuals resident in malaria-endemic regions show a high prevalence of seroreactivity to antigen Pf332 repeat sequences. The mean antibody concentrations were significantly higher in donors from Liberia, Madagascar and Gambia compared with Thai and Colombian donors, probably reflecting the higher degree of exposure in the African regions. Although the levels of such antibodies in individual sera correlated well with the levels of antibodies to one Pf155/RESA repeat peptide, only a minor part of the peptide-reactive antibodies were cross-reactive between the two antigens. In Gambian children, the mean concentrations of antibodies reactive with Pf332 or Pf155/RESA peptides were significantly higher in children with severe than with mild malaria. Further longitudinal studies are needed to evaluate the capacity of Pf332 to induce potentially protective or harmful antibody responses. PMID- 8403521 TI - Experimental murine paracoccidioidomycosis: relationship among the dissemination of the infection, humoral and cellular immune responses. AB - The dissemination of Paracoccidioides brasiliensis cells to the heart, omentum/pancreas, spleen, liver and lungs, assessed by colony forming unit (CFU) counts, the levels of specific antibodies to this fungal agent (by ELISA), and the specific DTH reaction were studied in susceptible (B10.A) and resistant (A/Sn) mice. The animals were infected intraperitoneally with P. brasiliensis yeast cells and were evaluated 2, 4, 12 and 16 weeks later. The most remarkable differences between the two mouse strains were observed 16 weeks after infection, when B10.A mice displayed high numbers of CFU in all examined organs, except the heart, high antibody titres, and depressed DTH response. At this point, A/Sn mice presented low or absent CFU in all organs, low antibody titres and expressive DTH response. The CFU counts were shown to be a reliable parameter to discriminate susceptible from resistant animals. The fungal load in the most affected organs correlated with the antibody titres and was inversely correlated with the intensity of the DTH reaction. The patterns of immune response in this model mimic human paracoccidioidomycosis, in which high specific antibody levels and depressed DTH reactions are found in multifocal and severe forms of the disease. PMID- 8403522 TI - Ascaris reinfection of slum children: relation with the IgE response. AB - Total and Ascaris-specific serum IgE levels were measured in a group of 98 Ascaris-infected children from a slum area of Caracas, Venezuela, in whom the infections were eliminated by regular treatment for 22 months with the anthelmint Oxantel/Pyrantel ('Quantrel'). The children were re-evaluated at the end of the treatment programme, and then 8 months later, at which time reinfection was assessed. Total IgE levels at the beginning of the study were significantly higher in the children who became reinfected after treatment, compared with those who did not. The anthelmint treatment caused a significant decrease in the total IgE levels in most of the children, and after a period of 8 months without treatment these continued to decrease in the non-reinfected group, but increased again in the reinfected children. The reverse pattern was found for Ascaris specific IgE antibody levels, and in fact an inverse correlation was found between total and anti-Ascaris IgE levels. Striking associations were found between reinfection and high pretreatment values of total IgE, but low levels of specific IgE antibody. These data support the concept that specific IgE antibody may participate in the protection against helminthic infection, and suggest that the polyclonal stimulation of IgE synthesis caused by these parasites may reduce the effectiveness of such responses. The results also indicate that different individuals have varying propensities to respond polyclonally to the helminths, and this influences their resistance to infection. PMID- 8403523 TI - Novel human immunoglobulin heavy chain constant region gene deletion haplotypes characterized by pulsed-field electrophoresis. AB - Fifteen patients with selective IgG1 deficiency were screened for immunoglobulin H chain C region locus (IGHC) gene deletions and three deletion haplotypes were found: del G1, del G1-G4 and del G4. These haplotypes, together with four deletion haplotypes described by us previously (del G1 (NY), del G1 (VIT) del G1 G2 (NY) and del G2-G4 (HJE)), were further characterized using pulsed-field gel electrophoresis (PFGE) to determine the physical extent of the deletions. The MluI fragment sizes confirmed the deletions, although the deduced sizes of the most extensive deletions indicated that material had been inserted into the locus. PMID- 8403524 TI - Duodenal intraepithelial gamma/delta T cells and soluble CD8, neopterin, and beta 2-microglobulin in serum of IgA-deficient subjects with or without IgG subclass deficiency. AB - Expression of the gamma/delta T cell receptor (TCR) on CD3+ intraepithelial lymphocytes (IELs) was studied by two-colour immunofluorescence in duodenal tissue sections from healthy (n = 6) or infection-prone (n = 7) subjects with selective IgA deficiency (IgAD), and subjects (n = 4) with combined IgAD and IgG subclass deficiency. TCR gamma/delta+ IEL proportions in selective IgAD subjects (median 6.3%, range 1.0-41%) and in those with combined deficiency (median 4.5%, range 1.2-33%) were well within the range (0.3-38%) for histologically normal controls (n = 11), but the healthy IgAD subgroup tended to show raised TCR gamma/delta+ IEL proportions (median 13.6%) compared with the other two subgroups. Also the number of TCR gamma/delta+ IELs per intestinal length unit was relatively high (median 13.9/mm) in the healthy IgAD subjects, and significantly raised (P < 0.03) compared with controls (median 3.2/mm). Paired staining revealed that most TCR gamma/delta+ IELs in both selective IgAD (98%) and combined deficiency (99%) were CD8-, and a large fraction (median 84% and 63%, respectively) expressed the V delta 1/J delta 1-encoded epitope. The total number of CD3+ IELs (mostly CD8+) was similar to controls. IgAD subjects, and especially the healthy subgroup, had significantly increased serum concentrations of soluble CD8 (P < 0.0002), neopterin (P < 0.005), and beta 2-microglobulin (P < 0.007), which was similar to our previous observations in common variable immunodeficiency, and probably reflected stimulation of cell-mediated immunity. In addition, the increased TCR gamma/delta+ IELs might reflect a component of compensatory surface protection in the healthy IgAD subgroup. PMID- 8403525 TI - Mannan-binding protein--levels in plasma and upper-airways secretions and frequency of genotypes in children with recurrence of otitis media. AB - We have investigated a possible association between recurrence of otitis media and low concentrations of mannan-binding protein (MBP) in plasma and upper-airway secretions. The protein concentration was measured in plasma (n = 76), nasopharyngeal secretions (n = 83) and middle ear effusions (n = 73) from otitis prone children, children with less recurrence of acute otitis media, children with no previous history of acute otitis media, but suffering from secretory otitis media, and healthy children. Moreover, genetic polymorphisms associated with low MBP plasma levels were investigated in DNA from nasopharyngeal tonsils of 89 children with recurrence of otitis media. A wide range of MBP plasma concentrations was found. No statistically significant differences in MBP plasma concentration were observed between patients and controls. Nor was there any increased frequency of the genotypes associated with low MBP plasma concentrations. Thus, our results do not support the assumption that low concentration and/or MBP deficiency alone predispose to recurrence of otitis media in Caucasian children. MBP was detected in both nasopharyngeal secretions (1/175 of plasma level) and middle ear effusions (1/4 of plasma level), suggesting a role for the protein in the local mucosal immune defense system at these locations. In contrast, MBP was undetectable in 53 samples of mixed-saliva. PMID- 8403526 TI - Evidence that imidazol(id)ine- and sulphonylurea-based antagonists of cromakalim act at different sites in the rat thoracic aorta. AB - 1. Ring segments of rat thoracic aorta were suspended in organ baths to record isometric tension. Tissues were precontracted with K+ (20 mmol/L), and full concentration-relaxation curves constructed to cromakalim (0.01-30 mumol/L) in the absence and presence of increasing concentrations of glibenclamide, glipizide, tolbutamide (the sulphonylureas), alinidine (an imidazolidine), phentolamine (an imidazoline), and chlorpromazine (the phenothiazine derivative). Whereas the active sulphonylureas, glibenclamide and glipizide, displayed classical competitive antagonism, the remaining compounds (alinidine, phentolamine and chlorpromazine) caused shifts in the cromakalim concentration effect curves associated with a reduction in the slope and maximum response. 2. A single concentration of each antagonist was selected and the shift in the concentration-effect curve determined. The possibility that sulphonylurea and imidazol(id)ine antagonists act at different sites was tested using the concentration-ratio method for combined antagonists described by Paton and Rang (1965). The combination of alinidine and phentolamine (collectively called imidazol(id)ines) at a number of different concentrations (10-30 mumol/L) resulted in a concentration-ratio to cromakalim which was additive, suggesting a common site of action. Similar results were obtained when examining the interaction between two sulphonylurea compounds (glibenclamide and glipizide). However, the interaction between sulphonylurea (glibenclamide) and imidazol(id)ine (alinidine) produced concentration-ratios which were multiplicative, suggesting a different or additional site of action for compounds from these two groups. Results indicated that chlorpromazine was able to block cromakalim via an action at the same site where alinidine and phentolamine act.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403527 TI - The intrarenal site of calcitonin gene-related peptide degradation in the isolated perfused rat kidney. AB - 1. Calcitonin gene-related peptide (CGRP) is a potent vaso-active 37 amino acid peptide, typically elevated in plasma from patients with medullary thyroid cancer (MTC), but undetectable in the plasma of normal subjects. 2. The kidney is a major site for the clearance of exogenously infused CGRP but the intrarenal site of this clearance is unknown. Extra-organ clearance is also significant for CGRP, and whereas the site and mechanism of this degradation remain uncertain, the vasculature has been postulated as the most likely site. 3. The isolated perfused rat kidney (IPRK) was studied to (i) localize the intrarenal site of CGRP clearance and (ii) determine the contribution of the renal vasculature to the clearance of CGRP. The half-life of CGRP in the filtering IPRK was 63.9 +/- 4.5 min, whereas blocking of filtration by elevation of the perfusate osmolarity abolished the degradation. This suggests that (i) renal CGRP degradation occurs after glomerular filtration with intratubular metabolism and (ii) that there is no active CGRP degradation in the (glomerular) capillary endothelium. 4. These results do not support the theory that renal vascular endothelium plays a major active role in CGRP degradation. PMID- 8403528 TI - Five year prospective study on blood pressure and maximal oxygen uptake. AB - 1. The relationship between physical fitness (maximal oxygen uptake VO2max) and incidence of hypertension was investigated through a prospective study for a total of 16,525 human-years of observation. 2. This study involved 3305 Japanese males whose blood pressure (BP) was normal when they received their first physical examination before the age of 50. They were monitored from 1983 to 1988. The BP of 425 subjects was diagnosed as hypertension in the fifth year. 3. Fitness levels were divided into quintiles according to VO2max levels, and were compared with the changes of BP and relative risk of hypertension after adjustment for age, initial percentage of body fat (PFAT), initial BP, alcohol consumption, cigarette smoking status and familial history of hypertension. The increase in BP of subjects in the least fit group was higher than in any other group. Relative risk was calculated using a multiple logistic regression and was 1.9 x higher in the least fit group compared with the fittest group. 4. The subjects were classified into three groups: the improved VO2max group, the deteriorated VO2max group and the unchanged VO2max group. The increase in BP of the improved VO2max group was significantly lower than the other two groups after adjustment for changes in PFAT, age, initial PFAT, initial BP, fitness level, alcohol consumption, cigarette smoking status and familial history of hypertension. 5. It is concluded that low VO2max level is related to higher incidence of hypertension. An improved VO2max would therefore be able to prevent hypertension. PMID- 8403529 TI - Haemodynamic and humoral consequences of chronic infusion of vasopressin in conscious rats. AB - 1. To determine the long-term haemodynamic and humoral effects of arginine vasopressin (AVP), a chronic intravenous infusion of AVP was performed in conscious Wistar normotensive rats. 2. AVP (1, 10, 50 or 100 ng/h) or saline as a vehicle control was infused continuously into the right jugular vein at a rate of 1 microL/h using an osmotic minipump for 7 days. 3. As a result, significant elevations of systolic blood pressure were observed in association with increases in plasma AVP concentration. Significant decreases in heart rate were observed during infusion of 100 ng/h of AVP. Mean arterial pressures measured directly on the sixth day of infusion were significantly higher in the rats given 50 ng/h (125 +/- 3 mmHg) or 100 ng/h (125 +/- 2 mmHg) compared with control rats (117 +/- 2 mmHg). Intravenous injection of the V1 antagonist, d(CH2)5Tyr(Me)AVP, significantly reduced the elevated mean arterial pressure induced by 50 or 100 ng/h of AVP (-7 +/- 4 and -11 +/- 2 mmHg, respectively). Plasma renin and norepinephrine concentrations were not affected by AVP infusion, while plasma epinephrine concentration was lower in the rats given 100 ng/h of AVP. Intravenous infusion of AVP did not alter bodyweight, serum electrolytes or osmolality. 4. These results suggest that AVP has a long-term pressor effect which is attributable to its vasoconstrictor action in conscious rats. PMID- 8403530 TI - Anti-arrhythmic effects of a new calcium antagonist, monatepil, AJ-2615, in experimental arrhythmic models [corrected]. AB - 1. To characterize the anti-arrhythmic properties of a new calcium antagonist, monatepil [corrected], AJ-2615, the preventive effects of AJ-2615 were compared with those of the existing calcium antagonists, diltiazem and verapamil, in experimental models of arrhythmia. 2. AJ-2615 (0.1-3.0 mg/kg, i.v.) suppressed ventricular arrhythmias induced by adrenaline (10 micrograms/kg, i.v.) in rats. AJ-2615 (0.1 mg/kg per min for 2 min, i.v.) also suppressed atrial tachycardia induced by aconitine (0.01% aconitine solution) in rats. 3. In these activities, AJ-2615 was comparable to or more potent than diltiazem and verapamil which are widely used for the treatment of arrhythmia. 4. In pro-arrhythmic activity, AJ 2615 was less potent than diltiazem and verapamil. 5. These results suggest that AJ-2615 would be a safer anti-arrhythmic agent, with less proarrhythmic liability than diltiazem and verapamil. PMID- 8403531 TI - Effects of peptide histidine isoleucine on pancreatic exocrine secretion in anaesthetized dogs. AB - 1. The effects of peptide histidine isoleucine (PHI) on pancreatic exocrine secretion were investigated in preparations of the isolated and blood-perfused dog pancreas as compared with those of vasoactive intestinal peptide (VIP), secretin and glucagon. 2. Each peptide tested was injected intra-arterially (i.a.) as a single bolus. Graded doses of PHI (3-300 nmol/kg), VIP (1-100 nmol/kg) and secretin (0.01-0.3 nmol/kg) caused dose-dependent increases in the secretion of pancreatic juice and bicarbonate outputs, but had little effect on the protein outputs. Glucagon (0.1-10 mumol/kg) produced a bell-shaped dose response curve for the secretory rate, bicarbonate and protein outputs. 3. The secretory activity of 30 nmol/kg of PHI corresponded roughly to that of 80 pmol/kg of secretin, 9 nmol/kg of VIP and 0.6 mumol/kg of glucagon, respectively. Thus, based on administered dose, PHI was about 375 x less potent than secretin, 3 x less potent than VIP and 20 x more potent than glucagon. 4. The PHI- and VIP stimulated secretions were inhibited by a VIP antagonist, but not by a glucagon antagonist, SCH23390 (a dopamine D-1 antagonist), L-364718 (a cholecystokinin antagonist) or atropine. 5. Each peptide increased cyclic AMP concentration, but not cyclic GMP concentration, concomitant with the increase in pancreatic secretion. 6. From these results, it is concluded that PHI produces an increase in pancreatic secretion by acting on VIP-preferring receptors on the exocrine pancreatic gland of the dogs. This may be mediated at least in part through the increase of intracellular cyclic AMP concentrations. PMID- 8403532 TI - Proliferative effect of mevalonate metabolites other than isoprenoids on cultured vascular smooth muscle cells. AB - 1. Recent investigations revealed that isoprenoid compounds serve as key substances for cellular proliferation through post-translational modification. Previously we reported that tissues of spontaneously hypertensive rats (SHR) had a lower activity of isoprenoid biosynthesis when compared with the normotensive control rat (WKY). However, cultured vascular smooth muscle cells (VSMC) of SHR showed an enhanced growth rate. These findings led us to investigate further the effect of isoprenoid compounds on VSMC proliferation. 2. When the cells of WKY were stimulated with 5% fetal calf serum (FCS) in the presence of lovastatin, [3H]-thymidine incorporation decreased in a dose-dependent manner and was completely inhibited at 30 mumol/L. Exogenously added mevalonate showed a protective effect against lovastatin (81% protection at 0.1 mumol/L). 3. Fluoromevalonate (Fmev), an inhibitor of mevalonate-PP decarboxylase which converts mevalonate-PP into isoprenoids, showed a dual inhibitory effect. DNA synthesis was partially inhibited at 0.01-1 mumol/L, however at 10 mumol/L there was no detectable inhibition. The inhibitory effect was again observed at concentrations over 10 mumol/L. 4. In the presence of lovastatin and Fmev to block both HMG CoA reductase and mevalonate-PP decarboxylase, exogenous mevalonate dose dependently stimulated [3H]-thymidine incorporation induced by FCS. 5. These data suggest the positive effect of the initial mevalonate derivatives other than isoprenoid compounds on the proliferation of VSMC. PMID- 8403533 TI - Analysis of secretion and expression of platelet-derived growth factor in cultured endothelial cells from stroke-prone spontaneously hypertensive rat. AB - 1. We characterized the endothelial cell-derived growth factors of SHRSP and Wistar Kyoto rats (WKY), respectively and found that platelet-derived growth factor (PDGF)-B chain related growth factor constituted a major portion of the mitogenic activity of the conditioned media of endothelial cells from both animals. There were no remarkable qualitative differences between the endothelial cell-derived growth factors of SHRSP and WKY. 2. Northern analysis revealed that the expression of PDGF-B chain was 2-4-fold enhanced in cultured aortic endothelial cells of SHRSP. This enhanced expression of PDGF-B chain, which may be induced under chronic hypertensive conditions, is suggested to contribute to the increase in endothelial cell-derived growth factors reported in this animal. PMID- 8403534 TI - Identification of adenosine receptors in human spermatozoa. AB - 1. The effects of adenosine receptor agonists and antagonists on human sperm motility have been studied. Specific binding sites for adenosine and its analogues on human sperm were also investigated. 2. Agonists stimulated human sperm motility in a dose-dependent manner with a potency order of 5'-N ethylcarboxamidoadenosine (NECA, EC50 = 0.3 mumol/L) > 2-[p (carboxyethyl)phenylethylamino]-5'-N- ethylcarboxamidoadenosine (CGS-21680, EC50 = 10 mumol/L) > adenosine (EC50 = 100 mumol/L). 3. NECA-stimulated motility was competitively inhibited by various adenosine receptor antagonists. The potency order was 3,7-dimethyl-1-propargylxanthine > 8-(p-sulfophenyl) theophylline > xanthine amino congener. 4. The radioligand [3H]-NECA bound to sperm membrane in a saturable manner with a Bmax of 21.3 pmol/mg protein and equilibrium Kd of 4 mumol/L. Adenosine agonists and antagonists competed for [3H]-NECA binding with the same rank order of potency as for the stimulation of human sperm motility. 5. GTP gamma s inhibited 63% of specific [3H]-NECA binding with IC50 value of 11 nmol/L. This suggests that the [3H]-NECA binding sites may be coupled to one or more G proteins. 6. These results indicate the presence of adenosine A2 receptors on human sperm which are responsible for adenosine-mediated enhancement of sperm motility. PMID- 8403535 TI - Atrial and brain natriuretic peptide response to exercise in patients with ischaemic heart disease. AB - 1. The response of venous plasma natriuretic peptides (atrial natriuretic peptide, ANP, and brain natriuretic peptide, BNP) plasma cyclic guanosine monophosphate (cGMP) and vasoactive hormones to dynamic exercise has been studied in 16 subjects undergoing diagnostic exercise tolerance for ischaemic heart disease (IHD), and in five healthy control subjects. 2. In patients with IHD, plasma ANP increased 3-fold (mean 16 +/- 2.5 pmol/L pre-exercise, 51 +/- 11 pmol/L after exercise, P < 0.01). Increase in plasma BNP (10.5 +/- 1.6 pmol/L pre exercise, 13 +/- 2 pmol/L after exercise, P < 0.01) was proportionately much less than ANP but more sustained. In exercising normal subjects, plasma ANP levels doubled (P < 0.01) but there was no significant change in plasma BNP levels. 3. In patients with IHD, there was a significant correlation between levels of plasma ANP and BNP before exercise (r = 0.97, P < 0.001) as well as during exercise (r = 0.79, P < 0.001). 4. Hormone responses in patients with positive exercise tests did not differ significantly from those with negative tests. 5. Although resting levels of plasma ANP and BNP in IHD are correlated, the findings indicate different mechanisms of secretion. The low BNP/ANP ratio in response to acute dynamic exercise presumably reflects the predominance of ANP in pre secretory atrial stores. PMID- 8403536 TI - Expression and functional characteristics of the complement receptor type 2 on adult and neonatal B lymphocytes. AB - In vivo antibody synthesis against thymus-independent type 2 (TI-2) antigens such as type-specific polysaccharides of pneumococci is low or absent during the first 2 years of life. Recently, we described a role for CR2 in the in vitro antibody response of B cells of adults to the TI-2 antigen type 4 pneumococcal polysaccharide. In the present study a decreased expression of CR2 is described on cord blood B cells using HB5 and OKB7 anti-CR2 MAb. Crosslinking of HB5 anti CR2 antibodies on the B cell membrane leads to increases in intracellular calcium ([Ca2+]i) in both adult and neonatal B cells. In adult B cells, a synergism between CR2 and sIgM could be demonstrated on the level of calcium mobilization by occupying CR2 and crosslinking of sIgM with substimulatory concentrations of anti-IgM antibodies. This synergistic action between CR2 and sIgM could not be demonstrated in neonatal B cells. In addition, it is demonstrated that HB5 MAb cannot induce B cell differentiation in neonatal B cells, while adult B cells can be induced to differentiate into Ig-producing cells by this MAb. PMID- 8403537 TI - A de novo deletion in the C1 inhibitor gene in a case of sporadic hereditary angioneurotic edema. AB - A sporadic case of hereditary angioneurotic edema (HANE) is reported here. The patient was a 15-year-old girl who for 4 years had suffered recurrent episodes of urticaria-like erythema, followed by vomiting with abdominal pain. She was diagnosed as having Sjogren syndrome by results of sialography and serological studies, and moreover, it was also observed that the C1 inhibitor (C1-INH) activity in her plasma was very low during these episodes of urticarial erythema. Diagnosis of acquired angioneurotic edema (AANE) was excluded because the patient's plasma had no inhibitory effect on the C1-INH activity of normal individuals. Although neither of her parents had a history of HANE, we were able to show that the patient had HANE by Southern blot analysis using C1-INH cDNA as a probe. One of the patient's C1-INH gene alleles was revealed to have at least a 17-kb-length deletion including exons 5-8. Neither her parents nor her healthy brother showed any abnormalities on Southern blot analysis. The parent-child relationship in this family was confirmed by an HLA-typing study of all family members. PMID- 8403538 TI - An activated CD8+ T cell phenotype correlates with anti-HIV activity and asymptomatic clinical status. AB - We have examined the relation of cell surface marker phenotype to the anti-human immunodeficiency virus (HIV) function of CD8+ cells from individuals at various clinical stages of HIV infection. Multiparametric flow cytometry analysis demonstrated that the most significant changes in cell surface phenotype was found within the CD8+ cell subset expressing HLA-DR and CD28. These changes in the levels of CD28 and HLA-DR expression on CD8+ cells were found to be related to antiviral activity and have clinical relevance based on three pieces of evidence: (1) strong CD8+ cell antiviral responses were associated in infected individuals with high levels of HLA-DR+ and CD28+ subsets; (2) CD8+ cell anti-HIV activity in vitro resides predominantly in the separated CD8+ cell subsets that express HLA-DR or CD28; and (3) in longitudinal studies, CD8+ cell anti-HIV activity decreased as an individual progressed from a healthy state to an AIDS condition. Taken together, the data suggest that the CD8+ cell subset identified phenotypically as a CD8+ CD28+ HLA-DR+ cell is responsible for natural anti-HIV activity. Longitudinal studies should determine if alterations in this subset can be used as a prognostic indicator for disease progression. PMID- 8403539 TI - Increased expression of Fc gamma RI on isolated PMN from individuals of African descent. AB - Fc gamma R plays an important role in host defense, triggering and/or facilitating many immunologic responses. Of the three defined Fc gamma Rs, Fc gamma RI (CD64) is not known to be constitutively expressed on normal PMN. We report here that there is markedly increased expression of Fc gamma RI on the PMN of normal, healthy blacks, detected by binding of monoclonal antibody to this receptor. This may have significant implications when multiracial data are pooled in studies of receptor expression as markers of response to various chemotherapeutic agents. PMID- 8403540 TI - Selective infiltration of B cells committed to the production of monoreactive rheumatoid factor in synovial tissue of patients with rheumatoid arthritis. AB - B cells were directly cloned using EBV transformation fron the synovial tissue of patients with rheumatoid arthritis (RA), the objective being to investigate the B cell repertoire at the site of inflammation. The frequency of clones producing antibodies with rheumatoid factor (RF) activity was approximately 10% in those from the RA synovial tissue. Similar percentages of B cell clones from the peripheral blood of both RA patients and healthy controls also produced RF. However, almost all of these clones from the peripheral blood produced RF reactive not only with rabbit IgG or human IgG Fc but also with several other antigens (polyreactive). Only one of 654 clones (0.15%) from the RA peripheral blood produced RF specific to rabbit IgG or human IgG Fc (monoreactive). On the other hand, the frequency of clones producing monoreactive RF was approximately 30 times higher in RA synovial tissue. Furthermore, these B cells were activated in vivo to produce antibodies, since monoreactive RF was spontaneously produced from synovial tissue cells without the addition of B cell stimulators. No clones producing monoreactive RF were obtained from the synovial tissue of patients with osteoarthritis. These results suggest selective infiltration and/or proliferation of B cells committed to the production of monoreactive RF in RA synovial tissue. PMID- 8403541 TI - Inhibition of epidermal growth factor-stimulated hepatocytes proliferation by autologous sinusoidal mononuclear cells in rat liver. AB - Mononuclear cells isolated from sinusoid of rat liver by collagenase perfusion method showed a strong inhibitory effect on epidermal growth factor-stimulated proliferation of cocultured autologous hepatocytes, while mononuclear cells from peripheral blood and spleen did not. This inhibitory effect depended on the effector/target ratio. In a single-color flow cytometric analysis, about 25% of sinusoidal mononuclear cells consisted of asialo GM1+ natural killer cells, and there were relatively small numbers of CD3+ T cells and a few leukocyte common antigen-positive B cells compared to peripheral blood mononuclear cells. The natural killer cell activity of sinusoidal mononuclear cells was much stronger than that of peripheral blood mononuclear cells and splenic mononuclear cells. This activity was completely suppressed by the treatment with anti-asialo GM1 antibody plus complement. However, their inhibitory activity on proliferation of autologous hepatocytes was not affected by this treatment. These results suggest that the sinusoidal mononuclear cells, probably CD3+ T cells, play a role in regulation of autologous hepatocyte proliferation. PMID- 8403542 TI - T lymphocyte responses to antigens of gram-negative bacteria in pyelonephritis. AB - We showed previously that large numbers of T lymphocytes accumulate within a few days in the kidneys of rats with ascending pyelonephritis induced with Escherichia coli or Pseudomonas aeruginosa. CD4+ T cells propagated from the lesions exhibited MHC-restricted proliferative responses to formalin-fixed bacteria of the species used to induce infection. In the present study we investigated further the nature of the antigens responsible for the T cell proliferation and studied the ability of different bacterial strains and species to produce proliferative responses. We found that heat-killed bacteria were more stimulatory than formalin-fixed bacteria, and that soluble supernatants of heat killed organism were also effective. The stimulatory effects of supernatants were destroyed by trypsin and the responses were MHC-restricted. Twelve different E. coli strains, with or without characteristics of uropathogenicity in humans, were all highly stimulatory to T cells derived from a kidney infected with a single E. coli strain. Strains of Klebsiella pneumoniae, Enterobacter aerogenes, and Serratia marcescens--species of Enterobacteriaceae closely related to E. coli- were also stimulatory, whereas more distantly related bacteria--Proteus, Morganella, and P. aeruginosa--were not. T cells propagated from kidneys infected with P. aeruginosa responded to supernatants of this organism, but not to E. coli supernatants. We conclude that a protein antigen (or antigens) shared by strains of E. coli and related Enterobacteriaceae, but not by other gram-negative bacteria, produce MHC-restricted proliferative responses of CD4+ T cells that infiltrate rat kidneys infected with E. coli. PMID- 8403543 TI - Isolation of leukocyte response integrin: a novel RGD-binding protein involved in regulation of phagocytic function. AB - We have described previously an adhesive protein on neutrophils (PMN) which recognizes fibrinogen, fibronectin (Fn), von Willebrandt's factor, vitronectin, collagen, and synthetic peptides containing the Arg-Gly-Asp (RGD) sequence (Gresham et al., J. Cell Biol. 108, 1935-1943, 1989). We have called this oligospecific receptor the leukocyte response integrin (LRI). Engagement of LRI leads to both increased ingestion via PMN IgG Fc receptors and to adhesion and chemotaxis to certain extracellular matrix proteins. Now, we have purified an RGD binding receptor from DMSO-differentiated HL-60 cells (dHL-60) by peptide affinity chromatography which has the biochemical, immunologic, and functional characteristics of LRI. The purified protein contains two bands of 135 and 90 kDa under nonreducing conditions SDS-PAGE. Immunologic characterization of the dHL-60 RGD receptor showed that, by Western blot and ELISA, the lower M(r) band was recognized by mAb 7G2, raised against placental beta 3, which is known to inhibit LRI function. However, despite this functional and immunologic cross-reactivity with beta 3, the receptor was not recognized efficiently by a polyclonal antibody to placental RGD-binding proteins, predominantly alpha v beta 3. Moreover, polyclonal antibody raised to the dHL-60 receptor (Ab1) did not react with placental RGD-binding proteins. By immunoprecipitation or ELISA, we demonstrated that the purified RGD-binding receptor was not alpha IIb beta 3 or alpha v beta 3 and did not contain the integrin chains alpha 4, beta 2, or beta 7. Functionally, Ab1 totally inhibited Fn-stimulated ingestion by PMN. Moreover, Ab1 inhibited phagocytosis stimulated by the peptide KGAGDV, which is the most specific ligand for LRI currently known, and Ab1 inhibited the binding of KGAGDV-coated microspheres to PMN and monocytes. FACS analysis with Ab1 showed staining of monocytes, PMN, and lymphocytes but not platelets or erythrocytes. We conclude that LRI is a novel RGD-binding receptor which exists on leukocytes and which shares an antigenic epitope(s) with beta 3. This receptor recognizes multiple RGD containing ligands and can mediate signal transduction for adhesion, chemotaxis, and activation of increased phagocytic potential by PMN and monocytes. PMID- 8403544 TI - Lack of role of TGF-beta 1 in decreased lymphoproliferative response in HIV-1 infection. AB - We investigated whether reduction of the phytohemagglutinin (PHA)-induced proliferative response of lymphocytes from HIV type-1 (HIV-1)-infected (HIV+) individuals could be explained by overproduction of transforming growth factor beta 1 (TGF-beta 1), a strong inhibitor of T cell proliferation. PHA-stimulated PBMC from 40 HIV- and 42 HIV+ homosexual men from the Baltimore Center of the Multicenter AIDS Cohort Study (MACS) were studied using Northern blot analysis of expression of TGF-beta 1 mRNA and determining the effects of anti-TGF-beta 1 neutralizing antibody on PHA-induced proliferative responses. Compared to the HIV donors, HIV+ donors did not show increased expression of TGF-beta 1 mRNA in unstimulated or PHA-stimulated PBMC. Furthermore, a neutralizing antibody to TGF beta 1 did not reverse the decreased proliferative response of PBMC from HIV+ individuals to PHA or interleukin-2. These results indicate that TGF-beta 1 is not involved in T cell proliferation defects seen in HIV+ donors. PMID- 8403545 TI - Monocyte chemoattractant protein-1 (MCP-1) in inflammatory joint diseases and its involvement in the cytokine network of rheumatoid synovium. AB - Macrophages infiltrated into synovium play an important role in joint destruction in inflammatory joint diseases. In this study we focused on the production of monocyte chemoattractant protein-1 (MCP-1), a recently identified monocyte chemotactic protein, by inflammatory synovium. Synovial fluid (SF) from rheumatoid arthritis (RA), osteoarthritis, gout, and traumatic arthritis contained MCP-1. MCP-1 was produced in the synovium of patients with RA and other inflammatory joint disease in in vitro culture systems; differences in the amounts produced were not significant. Synovial MCP-1 production in RA was further investigated. Levels of MCP-1 were significantly correlated with levels of IL-1 beta, IL-6, and IL-8 in the culture supernatants of synovia from RA. Using immunohistochemical techniques, MCP-1 was detected in the lining and sublining cells and in the vascular endothelial cells of rheumatoid synovia. Rheumatoid synovia with active inflammation were stained more intensely by anti MCP-1 antibody than were those with weak or inactive inflammation. IL-1 beta and TNF-alpha stimulated the expression of MCP-1 mRNA and de novo MCP-1 synthesis by cultured synovial cells. These results suggest the production of MCP-1 by synovium of various inflammatory joint diseases. In rheumatoid synovium, a cytokine network involving MCP-1 and other proinflammatory cytokines (IL-1 beta, IL-6, IL-8, and TNF-alpha) contributes to the immunopathogenesis of RA. PMID- 8403546 TI - Activation of human neutrophils and monocytes induced by immune complexes prepared with cationized antibodies or antigens. AB - We have recently showed that soluble immune complexes (IC) prepared with cationic antibodies (catIC) induce high levels of neutrophil-mediated cytotoxicity against nonsensitized target cells. In the present work we extended our previous findings by studying the ability of catIC to induce different responses mediated by monocytes and/or neutrophils: monocyte cytotoxicity against nonsensitized target cells, chemiluminescence emission by monocytes and neutrophils, and elastase release from neutrophils. Our results showed that, in all cases, cell responses induced by catIC were markedly higher than those induced by control IC, indicating that cationized antibodies enhance IC ability to trigger phagocytic cell activation. A second aim of the present study was to analyze the effect of antigen cationization on IC properties. Interestingly, we found that all the phagocytic cell responses induced by IC prepared with cationized ovalbumin (OA) were significantly higher than those induced by IC prepared with untreated OA. Our results suggest that the charge of antibody and/or antigen constitutes a critical property that conditions the biological activity of IC. Furthermore, these findings support an important role of cationic antibodies and antigens in the development of inflammatory events associated with certain IC-induced diseases. PMID- 8403547 TI - Inhibition of Fc gamma receptors in the plasma of subjects with Down's syndrome. AB - Subjects with Down's syndrome have several immunological abnormalities. We examined the sera of 29 subjects with Down's syndrome for the presence of Fc gamma receptor blocking and for the presence of anti-ssDNA antibodies by EA rosette inhibition. Fifty-five percent of Down subjects had levels of inhibition above the upper limit of normality in comparison to 7% of normal controls. The finding that after polyethylene glycol precipitation of selected sera giving high levels of EA rosette inhibition there was a reduction or a disappearance of the EA rosette inhibition could indicate that the blocking factors detected behaved as immune complexes. Since almost all subjects with anti-ssDNA antibodies also had elevated values of EA rosette inhibition, a role for immune complexes eventually formed with autoantibodies in an Fc-mediated immunoregulatory system is suggested. PMID- 8403548 TI - The effect of intravenous gamma-globulin on the induction of experimental antiphospholipid syndrome. AB - Antiphospholipid syndrome (APLS) is characterized by anti-cardiolipin antibodies (ACA) and/or lupus anticoagulant, recurrent thromboembolic phenomena, thrombocytopenia, and recurrent fetal loss. Recently, we reported on the induction of experimental APLS by passive transfer of ACA to naive mice. In the current study we examined the effect of intravenous gamma-globulins (IVGG) on the obstetric complication of experimental APLS. After showing the binding of IVGG to mouse and human ACA (e.g., the existence of natural anti-idiotypic autoantibodies to ACA) we infused 36 micrograms of IVGG to the tail vein of mice in which experimental APLS was induced by passive transfer of monoclonal mouse ACA (CAR). Mice treated with IVGG had significantly less fetal resorptions when mated, in comparison to untreated mice (5-13 +/- 6% vs 25 +/- 17%). The best results were achieved when IVGG was given 2 days after induction of the disease (on Day 6 of pregnancy). It seems that IVGG may be employed as a good adjunct to any therapy of APLS. PMID- 8403549 TI - From HIV infection to AIDS: are the manifestations of effective immune resistance misinterpreted? AB - Viewing the immune system as part of an adaptive communication network, rather than merely a coalition of effectors, we argue that the alterations in the immune system that occur in HIV-infected patients, generally considered to be "abnormalities," cannot be attributed directly to deleterious effects of the virus on CD4+ T cells or other particular cells. Rather, many of the functional changes that occur during the asymptomatic phase reflect a normal mode of immune resistance to chronic infection, different from the "stereotypic" immune response, whereby patterns of signals are recognized and classified and evoke selective activities. The relative stability of the virus-host relationship in this phase involves a degree of mutual adaptation. However, an excessively perturbed microenvironment is the core of unstable cellular organization in which the resistance to infection gradually deteriorates. We suggest that this is due to "overadaptation" of lymphocytes and accessory cells to the infectious agent(s). We further speculate that a key factor underlying this process is a reduced rate of replacement of CD4+ T cells, which are sequestered at the sites of infection, by fresh unprimed or memory T cells. Direct and local viral effects are amplified and propagated by "affected" cells, which are not necessarily infected. The collective profile of gene expression in various types of affected cells might adequately reflect tissue organization and the overall functional status of the immune system and thus could serve as a guide to therapy. This would require collection of a more extensive array of immunologic data than is now gathered, and novel approaches to analyzing such data. PMID- 8403550 TI - Alterations in C3, C4, factor B, and related metabolites in septic shock. AB - Forty subjects consisting of (i) patients with septic shock, (ii) patients with sepsis without shock, (iii) critically ill non-septic patients, and (iv) normal controls were studied for plasma C3, C4, factor B, iC3b, C4d, Bb, and SC5b-9 levels in an attempt to profile the pattern of complement activation in sepsis. Significant decreases in plasma C3 and C4 concentrations were observed in septic shock patients compared to normal as well as to critically ill subjects. When adjusted for illness severity, levels of C3 but not C4 showed significant differences between septic shock and critically ill patients. Concentrations of the complement metabolite Bb as well as the Bb to factor B ratio were significantly increased in septic shock patients. SC5b-9 levels were significantly higher in septic shock patients compared to normal as well as to critically ill subjects. No increases in C4d or iC3b were observed, but the iC3b to C3 ratio was higher in septic shock patients compared to that in normal subjects. In septic shock patients, there were significant correlations between SC5b-9 and Bb (P = 0.0213) concentrations and between C3 and Factor B concentrations (P = 0.0041). SC5b-9 levels also were correlated with lactate levels in septic shock patients (P = 0.0091). These findings demonstrate a metabolite pattern consistent with primarily alternative and terminal pathway activation in septic shock patients. Decreases in C4, a classical pathway component, may not be specific for septic shock and do not appear to be necessarily due to significant complement consumption/activation. PMID- 8403551 TI - Artemisinin and its derivatives enhance T lymphocyte-mediated immune responses in normal mice and accelerate immunoreconstitution of mice with syngeneic bone marrow transplantation. AB - Artemisinin and its derivatives, dihydroartemisinin and sodium artesunate, enhanced DNA synthesis of mouse spleen cells that had been activated with alloantigens or T cell mitogen Con A, but not B cell mitogen LPS, both in vitro and ex vivo. In vivo experiments showed that DTH response and antibody response against sheep erythrocytes were augmented in sodium artesunate-treated animals. In addition, sodium artesunate, a representative of these compounds, increased IL 2 production from mouse splenocytes stimulated with Con A. Because of the long term T cell function deficiency in mice after bone marrow transplantation, the effect of sodium artesunate on the immune reconstitution was assessed. The results demonstrated that 10 mg/kg sodium artesunate significantly accelerated the immune reconstitution in mice after syngeneic bone marrow transplantation. These data suggested that artemisinin and its derivatives appeared to promote T cell function selectively, and these compounds had a potential application for the recovery of immune function. PMID- 8403552 TI - Abnormalities in T cell interactions with the extracellular matrix proteins in a patient with Wegener's granulomatosis. AB - T cell interactions with the extracellular matrix proteins and cultured human endothelium were studied in a patient with Wegener's granulomatosis and other cases of vasculitis. Markedly enhanced costimulation of T-lymphoproliferative responses mediated by collagen and fibronectin were found in patients with severe forms of vasculitis, particularly with necrotizing changes. In addition, enhanced adhesion to collagen IV was found in the Wegener patient. T cell adhesion to resting and inflamed endothelium varied from normal to increased. PMID- 8403553 TI - Cocaine alters the respiratory burst and phagocytic activity of murine macrophages. AB - The effects of cocaine on the respiratory burst (RB) and the phagocytic activity of murine macrophages (M phi) were studied. C57BL/6 mice were injected intraperitoneally with various concentrations of cocaine or appropriate vehicle. Peritoneal and alveolar M phi were isolated from mice and cultured in vitro. Both alveolar and peritoneal M phi from cocaine-exposed mice exhibited an increase in the RB when compared with M phi from saline-injected controls. This increase in the RB was apparent 60 min after injection and persisted for at least 48 hr. Injection of the cocaine metabolites, ecgonine methyl ester hydrochloride and ecgonine hydrochloride, did not affect the RB. The increase in the RB was correlated with an increase in phagocytosis of opsonized zymosan in vitro. However, an in vivo phagocytic assay using mice injected ip with cocaine, which was subsequently followed by sheep erythrocytes, demonstrated the opposite effect. Fewer peritoneal M phi from cocaine-injected mice were observed with ingested erythrocytes. These data underscore the complex effects of cocaine on M phi functions. PMID- 8403554 TI - Expression of IL-2 receptor subunit p55 (CD25) on normal human lymphocytes. AB - Unstimulated peripheral blood lymphocytes of normal Saudi Arabian males were examined for the expression of IL-2 receptor subunit p55 (IL-2Rp55) by two-color flow cytometric analysis using monoclonal antibodies. Eighteen percent of the normal lymphocytes were positive for this marker. Approximately 84% of the IL 2Rp55+ cells were CD3+ T cells of which CD4+ and CD8+ T cell subsets constituted 75 and 13%, respectively. CD19+ B cells made up 12% of the IL-2Rp55+ lymphocytes. In the pan T cell population 20% cells were IL-2Rp55+, whereas among the B cells 16% cells were IL-2Rp55+. In the CD4+ and CD8+ T cell population IL-2Rp55+ cells were 29 and 7% respectively. In comparison, normal Caucasian male population had significantly higher proportions of IL-2Rp55+ lymphocyte subsets, the total IL 2Rp55+ lymphocytes being 33 +/- 10%. However, except for CD8+ T cells, there was no difference in the mean phenotypic values of IL-2Rp55+ cells between the two populations. The present findings clearly show a significant expression of IL 2Rp55 (CD25) on peripheral blood lymphocytes in normal, healthy Saudi Arabian blood donors and Caucasians. It is suggested that the baseline values of IL 2Rp55+ lymphocytes must be established for the local normal population before the expression of this marker is implicated either as an active immune response or as an immune disorder. PMID- 8403555 TI - Anti-CD3-activated T cells from chronic nonviremic HBV carriers are hyperreactive to monocytic accessory signals. AB - We analyzed T cell responses through the CD3 activation pathway in a group of chronic HBV carriers. PBMC stimulated with the mAb OKT3 showed higher proliferative response in HBV-DNA(-) carriers compared to HBV-DNA(+) carriers and to controls. In contrast, no differences in proliferative responses were observed between HBV-DNA(-) carriers and controls in cell cultures stimulated with immobilized 64.1 mAb (SPB-64.1) which induces proliferation in the absence of monocytes. We further examined T cell responses in the presence of monocytes and their soluble factors to immobilized OKT3 mAb (SPB-OKT3). Purified T cells did not proliferate to SPB-OKT3. When autologous monocytes were added, higher proliferative response, IL-2 production, and IL-2 receptor expression were observed in HBV-DNA(-) carriers than in controls. An enhanced cell proliferation was also obtained when monocyte supernatants were added to T cells cultured with SPB-OKT3. Moreover, when IL-6 alone or combined with IL-1 was added to SPB-OKT3 stimulated T cell cultures, a significantly higher increase in T cell proliferation was detected in HBV-DNA(-) carriers. Our results thus show a T cell hyperreactivity to accessory signals from monocytes (mainly IL-6) in HBV-DNA(-) carriers, that is probably related to an ongoing viral clearance. PMID- 8403556 TI - An excess of dietary iodine accelerates the development of a thyroid-associated lymphoid tissue in autoimmune prone BB rats. AB - Previous studies have shown that dietary iodine enhances the severity and incidence of focal thyroiditis in autoimmune BB rats and OS chickens. However, which lymphoid cells are involved in the development of the iodine-induced focal thyroiditis and what the consequences are for the anticolloid antibody production have not been studied in detail. We therefore performed a study in which 3-week old female BB rats were kept on either an enriched iodine diet (EID; iodine intake, 100 micrograms iodine/day) or a normal iodine diet (NID; iodine intake, 7 micrograms iodine/day) for a period of 18 weeks. The development of the focal thyroiditis was immunohistologically studied. Immunohistological data were compared to the thyroid hormone status and anti-colloid antibody production. Our data confirm that a high dietary iodine intake results in an accelerated development of the focal lymphoid cell infiltrates in the thyroid of the BB rat. After 12-18 weeks of an EID 50% of the BB rats developed these infiltrates. Our data additionally show that: (a) the process starts with increases in the number of infiltrating MHC class II-positive dendritic cells and a clustering of these cells with T cells, B cells, and some macrophages and (b) the focal infiltrates are highly organized and consist of central B cell follicle-like structures surrounded by rims and areas of T cells. The architecture of the focal thyroiditis is hence very similar to mucosa-associated lymphoid tissue and secondary lymphoid organs (spleen and lymph node). Only minor signs of thyrocyte destruction were observed. We therefore consider the term "thyroiditis" as inappropriate and prefer the term "thyroid-associated lymphoid tissue." Since the thyroiditis component was small, it is also not surprising that the BB rats on the EID remained euthyroid. The presence of the thyroid-associated lymphoid tissue in the BB rats was positively correlated to the presence of anti-colloid antibody in the serum of the BB rats. We speculate that the dietary iodine might have direct effects on cells of the immune system or on cells forming the microenvironment of lymphoid tissue (reticulum cells). A role for highly iodinated thyroglobulin in the accelerated development of thyroid-associated lymphoid tissue is also possible. PMID- 8403557 TI - Immunohistological demonstration of transforming growth factor-beta isoforms in the skin of patients with systemic sclerosis. AB - We investigated the expression of transforming growth factor-beta (TGF-beta) isoforms in involved and uninvolved areas of skin from patients with systemic sclerosis (SSc) and normal controls. Paraffin-embedded skin specimens were stained for TGF-beta 1, TGF-beta 2, and TGF-beta 3 using the avidin-biotin peroxidase technique. TGF-beta 2 was expressed intensely in the extracellular matrix of the skin biopsies obtained from involved areas of patients with SSc, in contrast to the uninvolved areas and normal individuals. TGF-beta 2 was deposited throughout the entire dermis and also in a linear fashion along the dermoepidermal junction and in the perivascular areas of SSc-involved skin. Although infiltrating mononuclear cells were not present in great numbers, they did not stain for TGF-beta 2. TGF-beta 1 and TGF-beta 3 were not expressed in the extracellular space in either patients or normal controls, except in rare cases. The cellular staining which was observed for all three isoforms did not differ between involved and uninvolved skin and normal controls. The finding of increased deposition of TGF-beta 2 in the involved skin of patients with SSc implies that it may be involved in the pathologic fibrotic process. PMID- 8403558 TI - Hydrocortisone regulation of interleukin-6 protein production by a purified population of human peripheral blood monocytes. AB - The direct effect of the endogenous glucocorticoid (GC) hydrocortisone (HC) on interleukin-6 (IL-6) production was examined using purified populations of human peripheral blood monocytes (Mo). Lipopolysaccharide (LPS) induced IL-6 production by Mo in a dose-dependent fashion. IL-6 was detected in Mo supernatants as early as 2 hr after stimulation, with peak IL-6 production observed by 16 hr. Simultaneous addition of HC and LPS resulted in a significant decrease of IL-6 production 4 hr after LPS treatment, with maximum inhibition observed at 16-24 hr. An attenuation of the inhibitory effect of HC occurred with greater concentrations of LPS and with the delay of HC addition until after LPS. However, there was no correlation between the quantity of IL-6 produced by Mo and the level of HC inhibition. The inhibitory effect of HC was greater if LPS, rather than IL-1 beta, were used as a stimulus to induce IL-6 production. The EC50 of LPS-induced IL-6 production by HC was 2.0 x 10(-7) M. The inhibitory effect of HC on LPS-stimulated IL-6 production was GC specific and receptor mediated because: (i) equivalent inhibition was not observed with other endogenous steroids and (ii) equimolar amounts of the GC antagonist RU 486 blocked the GC-mediated effect. PMID- 8403559 TI - Prevention of runting and cachexia by a chimeric TNF receptor-Fc protein. AB - Tumor necrosis factor alpha (TNF) is an important mediator of septic shock and cachexia. A soluble form of the human type 2 TNF receptor, constructed by joining the Fc region of human IgG1 to the TNF receptor, prevents weight loss in nude mice bearing a TNF-secreting tumor. This soluble receptor was also used to treat TNF transgenic mice which were runting and died before reaching reproductive age. After continuous treatment with soluble TNF receptor, the TNF transgenic mice grew to normal size and reproduced. Thus, soluble TNF may be useful in counteracting the detrimental systemic effects of TNF in a clinical setting. PMID- 8403560 TI - Rat adenocarcinoma 13762 expresses tumor rejection antigens but tumor-bearing animals exhibit tumor-specific immunosuppression. AB - Rat adenocarcinoma 13762 was adapted to continuous growth in culture and used in a variety of experiments to investigate the immune response to inoculation of animals with replication-defective tumor cells. The results demonstrate that 13762 cells express tumor-specific tumor rejection antigens that elicit protective immunity to tumorigenic challenge. By several criteria there is no apparent humoral component of the anti-tumor immunity; however, anti-tumor immunity is characterized by nylon-wool nonadherent spleen T cells. Anti-tumor T cells demonstrate tumoricidal activity in local adoptive transfer assays and are not found in spleens of naive animals or animals immunized against either nontumorigenic Rat 1 cells or a syngeneic fibrosarcoma. Despite the expression of tumor rejection antigens 13762 tumor, and the demonstrable ability of injection of irradiated tumor to induce anti-tumor immunity, tumors elicited in unimmunized syngeneic animals grow progressively. The reasons for growth of antigenic tumor are unknown but are shown not to be due to defective antigen expression in 13762 tumor since, in addition to being able to elicit T cell immune response in immunized animals, 13762 tumor expresses MHC Class I molecules and can be a target for allogeneic T cell recognition in vitro. These data suggest that in tumor-bearing animals an effective anti-tumor immune response is either not initiated or down-regulated. Since animals bearing 13762 tumors can be immunized against an unrelated syngeneic sarcoma, can produce humoral responses to several protein antigens, and can produce delayed type hypersensitivity response against dinitrofluorobenzene, the immune response to 13762-induced tumors appears specifically suppressed. In support of this contention, 13762 cells express high levels of transforming growth factor beta 1 in vitro which is postulated to impact upon the nascent anti-tumor immune response. PMID- 8403561 TI - Clinical immunology: guidelines for its organization, training, and certification. Relationships with allergology and other medical disciplines. A WHO/IUIS/IAACI report. World Health Organization. International Union of Immunological Societies. International Association of Allergology and Clinical Immunology. PMID- 8403562 TI - Effect of nifedipine on oxidative damage of erythrocytes in Plasmodium berghei infected mice. AB - It is known that the calcium channel blocker (CCB), nifedipine, can inhibit phagocyte oxidative burst in Plasmodium berghei-infected mice. The extent of immunopathological changes as seen by the course of infection and membrane lipid peroxidation in nifedipine-treated mice was examined in comparison with untreated mice at different parasite loads. The glutathione antioxidant system was also studied in these animals to assess its capacity to neutralize reactive oxygen species (ROS) in infected erythrocytes. The survival period of nifedipine treated, infected mice decreased significantly. It was observed that the accumulation of reduced glutathione was greater and the decrease in glutathione peroxidase activity was less marked in drug-treated animals, suggesting better protection of the parasites against oxidative injury. The accumulation of the lipid peroxidation product, malonyldialdehyde was significantly lower in nifedipine-treated animals at all parasitemia levels studied, indicating decreased ROS generation and parasite damage. These observations reveal the shortcomings of using CCB to reverse the chloroquine resistance in malaria as this would minimize oxidative damage of parasitized red cells and phagocyte mediated parasite killing. PMID- 8403563 TI - Decreased blood TcR gamma delta+ lymphocytes in AIDS and p24-antigenemic HIV-1 infected patients. AB - The significance of blood TcR gamma delta+ lymphocyte level was evaluated in the context of immunodeficiency and infections in 209 HIV-1-infected patients. Blood TcR gamma delta+ lymphocyte values were found higher in patients belonging to the CDC group II/III than those in the CDC groups IV C1 and IV D (P < 0.001) and P < 0.01, respectively). TcR gamma delta+ lymphocyte counts were lower in patients with oral candidiasis (P < 0.01), and in association with pneumocystosis or toxoplasmosis (P < 0.001). In 81 patients with a detectable HIV-1 p24 antigenemia, TcR gamma delta+ lymphocyte counts were lower than those in nonantigenemic patients (P < 0.001). In the CDC II/III group, p24-antigenemic patients exhibited lower TcR gamma delta+ cell counts than those in patients without antigenemia (P = 0.06). Data suggest that depletion of the TcR gamma delta+ lymphocyte subset characterizes HIV-1-infected patients with oral candidiasis, pneumocystosis, toxoplasmosis, and/or HIV-1-antigenemia. PMID- 8403564 TI - In search of nursing's history. PMID- 8403565 TI - Predictive value of small crescents in IgA nephropathy: analysis of four patients showing a deteriorated renal function during a long follow-up period. AB - Four patients with IgA nephropathy developed a chronic renal failure during a long follow-up period ranging from three to 8.5 years (mean 6.4 years). All patients showed a normal renal function, normal blood pressure and mild proteinuria at the time of the first renal biopsy. The first biopsies showed focal mesangial proliferation with small cellular crescents in a small percentage of the observed glomeruli. No case showed sclerotic changes in the interstitium and vessels. In contrast, at the second biopsies, all of them exhibited a deteriorated renal function, hypertension and massive proteinuria. The second biopsies revealed marked sclerotic changes in the glomeruli, interstitium, and vessels with significant focal segmental glomerular sclerosis and adhesions. Since no established factors predisposing the patients to chronic renal insufficiency had been observed at the time of the first biopsy, it was suggested that small crescents, even if focal, should be regarded as indicating an unfavorable prognosis. PMID- 8403566 TI - Deficiency of IgG subclass antibody response to tetanus toxoid associated with high serum IgA levels in IgA nephropathy. AB - In order to investigate IgG subclass response in IgA nephropathy (IgAN), 20 patients and 20 age and sex matched controls were systemically immunized with tetanus toxoid (TT). Nineteen/20 controls and 19/20 IgAN made a serum IgG anti-TT response of similar magnitude. However, significantly more patients with IgAN had undetectable amounts of at least one IgG subclass antibody to this antigen than controls (7/19 IgAN, 1/19 controls, p < 0.05). All individuals with an IgG subclass anti-TT deficiency lacked IgG1 and/or IgG4. Two patients made no IgG3 anti-TT (as well as no IgG1 and IgG4 anti-TT) but all individuals who responded to TT made IgG2 anti-TT. Total IgG subclass levels in IgAN did not differ from controls and no patient with IgAN had a total IgG subclass deficiency. Total serum IgA was significantly raised in IgAN (p < 0.002) and 6/7 IgAN with an IgG anti-TT subclass deficiency had a serum IgA level of over 3.2 milligrams compared to only 2/12 IgAN with no IgG subclass anti-TT deficiency (p < 0.01). The association of high serum IgA levels with IgG subclass deficiency to TT may be due to an abnormality in switching from IgG to IgA production in IgAN, or a manifestation of a defect of immunoregulation analogous to that proposed in IgA deficiency. PMID- 8403567 TI - An in situ hybridization study of herpes simplex and Epstein Barr viruses in IgA nephropathy and non-immune glomerulonephritis. AB - Renal tissues from forty cases of IgA nephropathy, 20 Singapore and 20 British patients, and 38 patients with non-immune glomerulonephritis were studied by in situ hybridization using DNA probes for the herpes simplex virus (HSV) and Epstein Barr virus (EBV). Two Singapore patients with IgA nephropathy showed HSV antigens in the glomerular mesangium, with one of them having EBV coinfection. In the control non-immune glomerulonephritis cases, 4 patients had viral antigens; 2 with EBV, one with EBV and HSV coinfection, and one with HSV alone. The study indicates that in some cases of IgA nephropathy and in non-immune nephropathy there may be coincidental rather than causal persistent infection by the herpes group of viruses. PMID- 8403568 TI - Hepatitis B virus associated membranous glomerulonephritis in children- experience in Hong Kong. AB - A retrospective review was undertaken on all renal biopsies performed in Chinese children under 13 years of age in Queen Mary Hospital, Hong Kong from 1980 to 1990. Of 172 renal biopsies, 18 cases were diagnosed to be membranous glomerulonephritis. All were associated with hepatitis B virus infection. Fourteen patients presented with, and 3 others developed later nephrotic syndrome. Hematuria was a common feature (83%). Complement C3 was low in 5 cases and they became normal later. At last observation, 10 patients (59%) remitted, usually within 3 years. One had persistent proteinuria and 5 had nephrotic syndrome. One patient developed end-stage renal failure 12 years after onset of illness. All three female patients completely remitted. Of the 15 boys, 50% remitted whether they received steroid treatment or not. Two patients received interferon alpha-2a therapy. One achieved remission but HBsAg and HBeAg persisted. The other had transient seroconversion and clinical improvement but nephrotic syndrome returned after stopping treatment. PMID- 8403569 TI - Increased release of tumor necrosis factor-alpha by monocytes from patients with glomerulonephritis. AB - The release of tumor necrosis factor-alpha (TNF) was measured in supernatants of cultured peripheral blood monocytes (PBM) that were obtained from patients with glomerulonephritis (GN) and healthy controls. Spontaneous and lipopolysaccharide (LPS)-induced TNF release were significantly higher in PBM isolated from patients with IgA nephropathy (IgA N) and membranous nephropathy (MN) as compared to normal controls. These biological activities were neutralized by a specific antibody to TNF. In patients with IgA N and MN TNF levels did not correlate with clinical disease activity. We conclude that in vitro GN PBM are hyperactive in their release of TNF, and that this enhanced release of TNF in vitro may reflect a secondary consequence of monocyte activation rather than a primary cellular defect in GN. PMID- 8403570 TI - Intravenous pulse cyclophosphamide in the treatment of type IV lupus nephritis. AB - A retrospective study compared two groups with type IV lupus nephritis with very similar activity and chronicity indices on renal biopsy. One group was treated with intermittent pulse cyclophosphamide (IPC), and the other was not. The IPC group demonstrated a greater decrease in serum creatinine at 6 months, 12 months and most recent follow-up intervals (p < 0.01, p < 0.05, p < 0.001). Reduction in proteinuria was similar in the two groups. Two of eight in the no-IPC group progressed to ESRD while only one of twelve in the IPC group developed ESRD. The findings suggest that IPC preserves renal function in Type IV nephritis. Furthermore it is suggested that a lower cumulative dosage than previously reported may reduce toxicity without a significant loss of efficacy. PMID- 8403571 TI - Cyclosporine in the treatment of lupus nephritis including two patients treated during pregnancy. AB - Although the combination of corticosteroids and intermittent pulse doses of cyclophosphamide has considerably improved the prognosis of lupus nephritis, there are still some unanswered questions about this regimen, in particular its use in pregnancy. As cyclosporine appeared to be effective in experimental models of lupus nephritis, some studies have been performed using this drug in patients with lupus nephritis. However, there was no mention of pregnancy in these patients. In view of the large experience with cyclosporine during pregnancy in renal transplant recipients and its established safety concerning teratogenicity, we decided to treat 5 young female patients having lupus nephritis with cyclosporine in combination with low-dose prednisone. Two of these patients were pregnant and both had successful delivery. During the follow-up period of 7-35 months there were no signs of flare-up of the lupus nephritis, except in one case where the patient accidentally discontinued the medication. In a second patient there was a slight increase of the serum creatinine level. Otherwise, the renal and immunological parameters improved or remained stable during the observation period. Hypertension developed in 3 cases. These preliminary results support the further evaluation of cyclosporine as an alternative to cyclophosphamide in the treatment of lupus nephritis, especially in young female patients with pregnancy or at high risk for pregnancy. PMID- 8403572 TI - Changes in partition of extracellular fluid volumes in anemic dialyzed uremic patients after partial correction of the anemia with recombinant human erythropoietin treatment. AB - The purpose of this work was to study the effects of correcting anemia on the distribution and partition of body fluids in dialyzed uremic subjects. We studied nine (7 m, 2 f) patients before and three months after the start of i.v. treatment with rHu-EPO, measuring total body water (TBW) with 3H2O, extracellular fluid volume (ECFV) with 35SO4 and plasma volume (PV) with 125I-SA. The intracellular water (ICW) and the interstitial fluid volumes (IFV) were derived by calculation from those measurements. The total blood volume (TBV) was calculated from the PV and the packed cell volume (PCV). Mean TBW, 482 +/- 45 (M +/- SD) ml/kg/bw and ECFV, 168 +/- 27.5 ml were significantly lower in patients than in nine matched normal controls, while the mean ICW (315 +/- 43 ml/kg) was similar. PCV before the start of rHu-EPO was 17.2 +/- 2.9% and had risen significantly to 31.3 +/- 4.8% (p = 0.000) after three months of therapy. Body weight (58 +/- 13 kg), TBW, ECFV and ICW did not change. TBV before rHU-EPO was 68.7 +/- 7.5 ml/kg and remained nearly unchanged, while PV fell significantly from 57 +/- 9 to 48 +/- 8 ml/kg (p < 0.025), with the calculated IFV rising from 111 +/- 25 to 127 +/- 27 (p = 0.000). The PV/IFV ratio decreased from 0.53 +/- 0.12 to 0.38 +/- 0.09 (p = 0.001). The decrease in PV/IFV ratio was paralleled by simultaneous increase in PCV in all but one patient.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403573 TI - Daily subcutaneous administration of recombinant human erythropoietin (rhEPO) in peritoneal dialysis patients: a European dose-response study. AB - In a prospective randomized open multicenter study, 107 anemic (Hct < = 28%) peritoneal dialysis (PD) patients were treated with s.c. rhEPO daily. The mean observation period was 299 days (range 14-479 days). Patients were randomly assigned to 3 groups with different initial doses: 5 U/kg (G5), 10 U/kg (G10), 20 U/kg (G20). Initial doses were maintained for at least 8 weeks unless the target Hct (30-35%) was achieved earlier. The weekly increase of Hct was significantly (p < 0.05) dose-dependent: 0.19% in G5, 0.5% in G10 and 0.94% in G20. In case of insufficient response (< 0.5% per week), the dose was doubled every 4 weeks. Final doses on achieving the target Hct ranged from 5 to 40 U/kg (median 20 U/kg). The dose was then reduced to 50% and adjusted individually. The median maintenance dose was 9.9 U/kg/day. No tendency towards higher blood pressure or intensification of antihypertensive treatment was observed. When rhEPO is administered daily, 10 U/kg/day (70 U/kg weekly) is the recommended starting dose. The need for higher doses used in unsatisfactory response, should lead to further examination to rule out iron deficiency and other reasons for non response. The median maintenance dose reported here is the lowest published in the literature for PD patients and seems to be linked to the daily injections. PMID- 8403574 TI - Is erythropoietin treatment safe and effective in myeloma patients receiving hemodialysis? AB - Still few data are available on efficacy and safety of recombinant erythropoietin (rEPO) in patients with myeloma and end-stage renal failure (ESRF); two such hemodialysed patients are reported in whom only partial response was observed, despite iron, folic acid supplementation and, in one case, high doses of rEPO (320 IU/kg/week). Despite improvement in well being and no need of further transfusion, hemoglobin did not reach 80 g/l. One patient developed recurrence 4 weeks after starting rEPO. Patients with ESRF and myeloma should benefit from rEPO but particular attention should be paid to marrow proliferation. PMID- 8403575 TI - Microcirculation of nailfold capillaries in chronic hemodialysis patients with and without diabetes mellitus. AB - The present study was intended to determine whether hemodialysis patients with arteriovenous fistula showed changes of microcirculation in the upper extremity. In addition, the study was to show whether such changes were correlated to the shunt parameters, such as the shunt flow volume, and whether the presence of diabetes mellitus had any additional influence. The study included 43 patients undergoing chronic hemodialysis; ten of the 43 patients were diabetics. Morphological and hemodynamic parameters of microcirculation were determined using quantitative capillaroscopy, flow volume in the brachial artery and shunt flow were measured by means of pulsed Doppler ultrasonography. While the morphological microcirculatory parameters did not show any differences between shunt arm and contralateral side, statistically significant differences were seen for resting blood cell velocity and for the time to peak capillary blood cell velocity after reactive hyperemia. For the latter a correlation to the shunt flow volume could also be demonstrated for nondiabetic patients. As expected, the time to peak blood cell velocity after reactive hyperemia was prolonged in both arms of patients with diabetes mellitus. Apparently, diabetes mellitus had a more pronounced effect on hemodynamic parameters than the Cimino shunt. This and the shunt flow will result in a Raynaud symptomatology. Other symptoms associated with chronic hemodialysis are mainly related to the duration of hemodialysis. PMID- 8403576 TI - In vitro solubilization of glomerular immune deposits after incubation with human IgG and IgA in membranous and IgA nephropathy. PMID- 8403577 TI - Cyclosporin A and proteinuria. PMID- 8403578 TI - Autoantibodies in vasculitis: current perspectives. PMID- 8403579 TI - Coexistence of Paget's bone disease and diffuse idiopathic skeletal hyperostosis in males. AB - The coexistence of Paget's bone disease (PBD) and diffuse idiopathic skeletal hyperostosis (DISH) of the spine is controversial and could have implications for the pathogenic mechanisms involved in these disorders. In order to assess the prevalence of DISH in patients with Paget's disease, a prospective controlled study was carried out in which roentgenograms were obtained from a group of 50 consecutive ambulatory patients previously diagnosed as having PBD (25 males and 25 females; mean age: 66.2 +/- 8.9 years) and these were compared with 50 age- and sex-matched subjects selected randomly from various categories of medical disorders excluding PBD. DISH was found in 12 of 50 Paget's patients, this corresponding to an incidence of 24%; 10 (83%) of these 12 subjects were males. In the control population the incidence of DISH was 6%-2 males and 1 female. This difference between the two groups was statistically significant (chi 2 test; p = 0.02). Apart from the male gender, which was clearly associated with the presence of DISH in the Paget's group (p = 0.019), no other biological variable or characteristic of PBD was linked to the presence of DISH. In conclusion, our data revealed that PBD patients have a significantly greater prevalence of DISH than non-PBD probands. It would seem that the genetic mechanism which is responsible for the susceptibility of PBD to osteoclast viral infection could be related to the pathogenia of DISH in a sex-linked manner. PMID- 8403580 TI - Intra-articular treatment with hyaluronic acid in osteoarthritis of the knee joint: a controlled clinical trial versus mucopolysaccharide polysulfuric acid ester. AB - In a single-blind, randomized clinical trial, both the efficacy and safety of hyaluronic acid (HA) were compared with that of mucopolysaccharide polysulfuric acid ester (MPA) in patients with osteoarthritis of the knee joint. Both agents were administered intra-articularly over six weeks. Patients received either seven injections of HA or 13 injections of MPA. Joint function, range of motion, severity of pain, the general condition of the bony structure and soft tissue of the joint area, and the global clinical efficacy and safety of the medication were assessed. The mean improvement in the modified total Larson rating score was 22% (SD = 28) after HA treatment and 7% (SD = 17) after treatment with MPA (analysis of variance: p = 0.02). This change was mainly caused by a reduction of pain. The onset of pain relief was more rapid in the HA group. The therapeutic effect increased in both treatment groups during the follow-up period. During this interval, lasting six months after the start of treatment, a further reduction of pain and an improvement of knee joint function could be observed. At the end of the study, 25 out of 33 (76%) patients in the HA group and 11 out of 24 (46%) patients in the MPA group were symptom-free or markedly improved (Chi square test: p = 0.02). Both agents were tolerated very well. PMID- 8403581 TI - Neurological manifestations of systemic lupus erythematosus: role of antiphospholipid antibodies. AB - Antiphospholipid antibodies (APL) are associated with venous and arterial thrombosis in SLE patients. Various thrombotic and non-thrombotic neurological manifestations have been reported in SLE but whether or not they are related to the presence of APL antibodies remains uncertain. To assess the possible association between neurological involvement in SLE and APL antibodies, IgG anticardiolipin antibodies (IgG ACL) were looked for using an ELISA technique in 92 consecutive SLE patients seen over a one-year period. Other APL determinations included VDRL and lupus anticoagulant (LAC) testing using APTT and the diluted thromboplastin time. Twenty-four SLE patients presented with neurological manifestations (40 episodes): 15/24 (62.5%) were found positive for APL antibodies (11 VDRL, 8 LAC, 7 ACL antibodies) versus 22/68 patients (32%) without neurological symptoms (p < 0.01). APL antibodies antedated neurological symptoms in 13/16 cases. Neurological manifestations were subsequently divided into 3 groups: thrombotic (n = 14), psychosis and convulsions (n = 15), miscellaneous (n = 10). No correlation was found between APL antibodies and any of the 3 subgroups. Among patients with neurological SLE, APL antibodies were present in two with valvular heart disease, as well as in seven with a history of either deep vein thrombosis, livedo reticularis or miscarriage. Among 7 patients with thrombocytopenia and neurological symptoms, 6 had APL antibodies. These data suggest that APL syndrome is associated with neuro-ophthalmological manifestations of SLE regardless of whether or not the mechanism of neurological involvement is thrombotic. SLE patients with APL antibodies may be at risk for future neurological manifestations. However, it is still questionable that APL positivity has definite therapeutic consequences. PMID- 8403582 TI - Bone mineral density in ambulant, non-steroid treated female patients with rheumatoid arthritis. AB - Bone mass measurements were performed in a group of 30 ambulant, non-steroid treated female patients with rheumatoid arthritis (RA) of relatively short duration (mean 4.9 years). The bone mineral density (BMD) of the lumbar spine and femoral neck was assessed by dual-energy x-ray absorptiometry (DEXA), and related to parameters of disease activity and severity. Lumbar BMD was within the range of normal while femoral BMD was decreased compared to age-matched controls. BMD values, expressed as the percentage of age-matched healthy controls (BMD%), were positively related to the body mass index and negatively related to the number of swollen joints, the erythrocyte sedimentation rate and the platelet count. No relation was found between the lumbar and femoral bone mass on the one hand and disease duration, number of disease modifying anti-rheumatic drugs ever used, Ritchie articular index, C-reactive protein, functional ability or radiological scores on the other. It is concluded that in ambulant non-steroid treated female RA patients lumbar bone mass as measured with DEXA is within the range of normal, while femoral bone mass is slightly reduced. Both lumbar and femoral bone mass are related to the body mass index and parameters of disease activity. PMID- 8403583 TI - D-penicillamine induced myasthenia gravis: clinical, serological and genetic findings. AB - Eight cases of D-penicillamine (DP) induced myasthenia gravis (MG) are presented. Seven patients were being treated for rheumatoid arthritis (RA) and one for scleroderma. The mean duration of DP treatment until the myasthenic symptoms developed ranged from 2-8 months. The DP dose reached 500 mg daily. It was found that the clinical and immunological findings were almost similar to those of idiopathic MG, but were less severe. All patients had increased titers of acetylcholine receptor antibodies in their sera. Discontinuation of D penicillamine resulted in the complete resolution of myasthenic symptoms after 2 6 months. One patient required ventilation, immunosuppressive therapy and plasma exchange. No association was found between DP related MG and the various autoantibodies tested. Immunogenetic analysis showed that three patients had HLA DR1, two HLA-DR3, one HLA-DR4 and one HLA-DR5. In conclusion, the clinical presentation of DP-induced MG seems similar to idiopathic MG. DP-related MG is relatively benign, although it sometimes can cause life-threatening muscle weakness requiring aggressive therapy. The relatively small number of patients included in this study, however, does not permit any firm conclusions regarding the HLA associations of DP-related MG. PMID- 8403584 TI - Detection and identification of antinuclear autoantibodies in the serum of normal blood donors. AB - The occurrence of antinuclear antibodies (ANA) in the serum of 485 healthy volunteer blood donors was assessed. Sixty two sera displayed nuclear immunofluorescence staining on Hep-2 cells using a polyvalent anti-Ig conjugate. In general, the titer of these antibodies was low (42/62 sera displaying a titer lower than or equal to 1:80). In only 23 sera were the ANA of the IgG isotype, which is the more disease-related immunoglobulin class of autoantibodies. In order to define the frequency of antibodies to extractable nuclear antigens and dsDNA within this population, sera were further analyzed by counterimmunoelectrophoresis. Western blot and the Crithidia luciliae assay. One serum displayed weak antids DNA reactivity; another serum had anti-SSA/Ro activity. On Western blot several patterns were found. They could not be identified with any of the available reference antisera. PMID- 8403585 TI - The significance of enthesopathy as a skeletal phenomenon. AB - Enthesophytes and enthesopathy, while easy to define, represent a phenomenon of unclear clinical significance. As the high frequency in skeletal populations suggests that enthesopathy may not be disease-specific, the nature of the reaction was assessed in 872 individuals from a representative skeletal population, subdivided into groups characterized by the presence or absence of rheumatoid arthritis, spondyloarthropathy, calcium pyrophosphate deposition disease and diffuse idiopathic skeletal hyperostosis (DISH). Achilles, plantar fascia, patellar and iliac crest entheses were examined for evidence of calcific overgrowth or "erosions." Enthesophytes were found to be a phenomenon of aging in individuals, and unrelated to the presence of inflammatory arthritis or DISH. The frequency increased with age, independent of sex or the site examined, plateauing in frequency after age 60. Enthesophytes in individuals under age 60 were usually unrelated to any underlying disorder. The absence of effect of underlying forms of arthritis on the frequency of enthesophytes at the patellar, Achilles and plantar sites suggests that mechanical factors outweigh the "enthesis calcifying" impact of such disorders as spondyloarthropathy, calcium pyrophosphate deposition disease and DISH. Individuals with rheumatoid arthritis, however, manifested a less severe iliac crest enthesial reaction, in keeping with the minimal reactive new bone formation characteristic of its erosions. Analysis of Achilles, plantar, and patellar enthesial reactions as a function of underlying inflammatory arthritis or DISH also revealed no significant variation with the underlying process. Cortical discontinuity at enthesial sites was a relatively infrequent phenomenon. While calcaneal discontinuities were originally thought to be erosive in nature, these observations suggest the possibility of tendon avulsion injuries. PMID- 8403586 TI - A double-blind study to determine the duration of action of flurbiprofen in a sustained release preparation. AB - This double-blind, placebo-controlled, parallel-group study proves that the duration of activity of a sustained-release preparation of flurbiprofen 200 mg covers a full 24 hour period. In a group of 24 patients with clinically active rheumatoid arthritis a statistically significant increase of pain was noted 37 hours after the last active dose. The increase in disease symptoms was inversely related to the plasma flurbiprofen levels and was rapidly reversed with one active dose of sustained release flurbiprofen. PMID- 8403587 TI - Is there a relationship between subclinical myocardial abnormalities, conduction defects and Ro/La antibodies in adults with systemic lupus erythematosus? AB - Thirty-three adult patients with systemic lupus erythematosus (SLE) were studied to determine whether there was an association between subclinical abnormalities of left ventricular function, conduction defects, and the presence of antibodies to Ro and/or La. Twelve patients had one or both of these antibodies. Conduction defects were present in 2, neither being currently positive for Ro or La antibody. There was no difference in abnormalities of left ventricular function between those with or without antibody. Valvular abnormalities were present in 11, 6 of whom had focal or diffuse thickening of one or more valves. Adults with SLE and anti-Ro and/or La antibody do not appear to be at particular risk of conduction defects. Subclinical left ventricular functional abnormalities were not associated with the presence of these antibodies. PMID- 8403588 TI - Patients with ankylosing spondylitis and healthy relatives do not show increased small intestinal permeability with the lactulose-mannitol test. AB - Small intestinal permeability was measured in 71 subjects: 26 (24 B27+) patients with ankylosing spondylitis (AS); 20 healthy first degree relatives (13 B27+); 6 patients with active Crohn's disease and 19 healthy controls. We determined the urinary excretion ratio of two ingested sugar probes, lactulose (10 g) and mannitol (0.5 g) by gas-liquid chromatography. The median lactulose/mannitol excretion ratio in AS patients (0.0099) and relatives (0.0090) was not significantly different from the median ratio in healthy controls (0.0095). HLA status or use of NSAIDs did not significantly influence the results. In patients with Crohn's disease, on the other hand, the median lactulose/mannitol ratio (0.021) was significantly increased in comparison to healthy controls (0.0095). Our results confirm that the lactulose-mannitol test can be used to demonstrate increased intestinal permeability in Crohn's disease. For patients with AS and their relatives the lactulose-mannitol test may not be sufficiently sensitive. Alternatively, significantly increased permeability may not occur in most patients with AS. PMID- 8403589 TI - Lymphoscintigraphic study in a case of rheumatoid arthritis-related lymphoedema. AB - Lymphoedema of the upper limbs is a rare extraarticular manifestation of rheumatoid arthritis (RA). Herein we report a patient with RA who presented two episodes of lymphoedema in the hands and forearms coinciding with flares of polyarthritis. Lymphoscintigraphy showed lymphatic-ectasia. The oedema improved with slow-acting drug treatment. PMID- 8403590 TI - An identical twin pair discordant for rheumatoid arthritis and ankylosing spondylitis. AB - Both rheumatoid arthritis (RA) and ankylosing spondylitis (AS) have an increased familial occurrence and each disease is associated with the inheritance of specific HLA antigens. We report a pair of identical twin brothers with discordant disease phenotypes: one developed AS at the age of 26, and the other developed RA at the age of 55. The twins possessed both of the disease susceptibility antigens HLA B27 and DR4. Differences in the twins' environmental exposure are discussed. PMID- 8403591 TI - Erythropoietin and the anemia of chronic diseases. AB - The red-cell mass is continuously adjusted to the optimal size for its function as an oxygen carrier by messages transmitted to the bone marrow from an oxygen sensor in the kidney. These messages are mediated by the hormone erythropoietin. Erythropoietin is a glycoprotein growth factor synthesized by cells adjacent to the proximal renal tubule in response to signals from a renal oxygen-sensing device, probably a heme protein (1). In the bone marrow, erythropoietin binds to and activates specific receptors on the erythroid progenitor cells (2). In the presence of this erythropoietin-receptor complex the progenitor cells continue their predestined development into mature erythrocytes. Erythropoietin was the first hemopoietic growth factor to be molecularly cloned in 1985 (3). Our understanding of the biology and physiology of erythropoietin has been considerably improved with the advent of recombinant human erythropoietin (rHuEpo). During the past 7 years, rHuEpo has undergone extensive testing in clinical trials. It has been approved for treatment of the anemia of chronic renal failure, both in progressive renal failure and endstage renal failure (ESRD). In these instances, the administration of rHuEpo has been used in effect as a substitutive therapy, since patients' erythropoietin levels are very low despite severe anemia, due to the failure of affected kidneys to produce adequate amounts of the hormone. However, the application of rHuEpo has now moved largely from the primitive indication of renal diseases, and the hormone is currently under study in a number of anemic states of different etiologies, even with relatively high serum erythropoietin levels. Among these, some of the best documented indications are the anemia associated with malignancies, either due to neoplastic bone marrow infiltration or to chemotherapy-related myelosuppression, the anemia of myelodysplastic syndromes and AIDS, the anemia of chronic inflammatory diseases, prematurity, and bone marrow transplantation (4). The purpose of this review is to provide a summary of our present knowledge regarding rHuEpo therapy for the anemia of renal failure. We provide some clues for the correct use of rHuEpo in the treatment of the anemia of chronic inflammatory diseases. In addition, we address a series of new issues in the attempt to better understand the relationship between erythropoietin and liver disease. PMID- 8403592 TI - Autoantibody production in Kawasaki syndrome. AB - Although a variety of autoantibodies are produced in patients with Kawasaki syndrome (KS), their specificities in many instances are controversial and their role in disease pathogenesis is undetermined. Autoantibody production was studied in 14 patients with Kawasaki syndrome (KS). Antibodies to myeloperoxidase (MPO), the dominant antigen responsible for perinuclear antineutrophil cytoplasmic antibody (pANCA) reactivity, were detected by ELISA in 73% of acute phase and 89% of convalescent phase KS specimens, in contrast to 4% of normal adult control subjects (p < 0.002 and p < 0.001, respectively). MPO and cytoplasmic antineutrophil antibody (cANCA) levels measured by ELISA were significantly elevated above levels for adult normal control subjects (p < 0.005 and p < 0.01, respectively), but not above recently ill childhood controls. Among patients who developed a positive ANCA, antibody titers tended to rise in serial specimens despite clinical improvement. Antibodies to myocardial muscle, cardiac perimysial connective tissue, nuclear antigens (ANA), and smooth muscle were also detected in some KS patients, but titers did not differ significantly from control patients. Autoantibody results were not predictive of patients with echocardiographic abnormalities. PMID- 8403593 TI - Macrophage activation syndrome and rheumatic disease in childhood: a report of four new cases. AB - A macrophage activation syndrome (MAS) developed in four children with chronic rheumatic diseases. The presentation included fever, hepatic and splenic enlargement, profound depression of blood counts, lowering of ESR, elevation of SGOT/PT and hypofibrinogenemia. The most characteristic sign of MAS was the presence in the bone marrow aspirate of well differentiated macrophages showing active haemophagocytosis with haematopoietic elements in their cytoplasm. Activation of the macrophage was also illustrated by high levels of monokines in the serum of 2 patients. This immuno-hematological process of unknown etiology can be triggered by ubiquitous events such as infections and treatment with anti inflammatory drugs. It is a potentially lethal complication which should be diagnosed rapidly, since administration of high-dose steroids with discontinuation of potentially toxic drugs can induce remission. Cyclosporin A was effective in two patients and may be of value in the management of the macrophage-activation syndrome. Its efficacy supports the central involvement of a T-cell dysfunction. It must be borne in mind that children with rheumatic diseases, especially the systemic form of juvenile chronic arthritis, are highly vulnerable to life-threatening macrophage activation, which appears to be more frequent than previously recognized. Very careful monitoring of apparently "innocent" drugs and intercurrent viral infections is thus required. PMID- 8403594 TI - A patient with systemic sclerosis, severe cytopenias and the simultaneous presence of anti-centromere and anti-SCL-70-antibodies. PMID- 8403595 TI - Colchicine and thrombopenia. PMID- 8403596 TI - Arthritis, cutaneous vasculitis and autoantibody multiplicity following thymothymomectomy for pure red cell aplasia. PMID- 8403597 TI - Development of persistent destructive arthritis in a patient with rheumatoid nodulosis in spite of an improvement in extra-articular symptoms. PMID- 8403598 TI - The genetics revolution and the role of the obstetrician. PMID- 8403599 TI - Immediate and long-term applications of technology. PMID- 8403600 TI - Reproductive health and genetic testing in the Third World. PMID- 8403601 TI - Prenatal diagnosis: choices women make about pursuing testing and acting on abnormal results. PMID- 8403602 TI - Prenatal genetic services: toward a national data base. PMID- 8403603 TI - Provider biases and choices: the role of gender. PMID- 8403604 TI - Feminist criticism of prenatal diagnosis: a response. PMID- 8403605 TI - Legal and regulatory issues surrounding carrier testing. AB - Genetic testing has arrived, probably earlier than patients and physicians need. For the moment, professional societies are taking the lead in monitoring the quality of physician education and laboratory services. The federal government will soon take over the role of monitoring the quality of genetic test kits themselves, but the most significant development will be the evolving physician patient relationship in the context of primary and prenatal care. As of 1992, it is probably not necessary for physicians to educate their patients about the availability of genetic tests unless there is a specific indication of genetic disease in the family. However, should a patient ask for such information or testing, today's physician would have a duty to know enough about the current status of carrier testing to be able to respond to the requests or make a proper referral. In addition, as the reliability of such tests increase and their costs decrease, physicians may arrive at a moment when some sort of patient education is required, at least for the most common disorders. The recent court decisions securing the right to abortion mean that patients will continue to have the moral and legal right to assert their privilege to plan their families and, where possible, to avoid genetic impairments in their children. Physicians will have the sensitive task of helping patients to achieve their personal goals regarding such family planning, while not overwhelming patients with confusing or frightening information. PMID- 8403606 TI - New reproductive genetics: political issues. PMID- 8403607 TI - Prenatal screening and its impact on persons with disabilities. PMID- 8403608 TI - Gonadotropin-releasing hormone and its analogues: from laboratory to bedside. PMID- 8403609 TI - Gonadotropin-releasing hormone agonists: treatment of endometriosis. PMID- 8403610 TI - Gonadotropin-releasing hormone agonists: utilization before hysterectomy. PMID- 8403611 TI - Use of gonadotropin-releasing hormone agonists before myomectomy. PMID- 8403612 TI - Treatment of uterine leiomyomas in perimenopausal women with gonadotropin releasing hormone agonists. PMID- 8403613 TI - Treatment of abnormal uterine bleeding with gonadotropin-releasing hormone analogues. PMID- 8403614 TI - Use of gonadotropin-releasing hormone agonists in the treatment of hyperandrogenism. PMID- 8403615 TI - Effects of gonadotropin-releasing hormone analogues on ovarian epithelial tumors. PMID- 8403616 TI - Use of gonadotropin-releasing hormone agonist to cause ovulation and prevent the ovarian hyperstimulation syndrome. AB - The physiologic basis and clinical applications of the use of GnRHa, rather than hCG, to induce the final stage of oocyte maturation and ovulation in gonadotropin treated cycles were reviewed. A single mid-cycle dose of GnRHa is able to trigger a preovulatory LH/FSH surge, leading to oocyte maturation and pregnancy in women undergoing ovarian stimulation for IVF/ET or induction of ovulation in vivo. The limited information currently available suggests there are similar pregnancy rates in patients treated with either GnRHa or hCG. The potential clinical advantages of GnRHa over hCG in gonadotropin-treated cycles include 1) the ability to titrate the amplitude and duration of the LH surge, 2) better control of luteal steroid hormone levels, 3) a higher implantation rate, 4) a lower rate of multiple pregnancy, and 5) a reduced risk of OHS. To date, the GnRHa regimen has been effective in preventing OHS in patients at high risk for having this complication. PMID- 8403617 TI - Utility of gonadotropin-releasing hormone agonists in programs of ovarian hyperstimulation with intrauterine insemination. AB - The GnRH agonists have practical and theoretic advantages for adjunctive use in ovulation induction. The IVF cycles demonstrate a decrease in the cancellation rate, an increase in the ease of scheduling, and an increase in the number of oocytes obtained per retrieval when GnRH agonists are employed. Other advantages, such as an improvement in the fertilization and cleavage rate, an increased length of the luteal phase, and an increased pregnancy rate, are suggested but not universally accepted. The utility of adding GnRH agonists to human menopausal gonadotropin-intrauterine insemination cycles is similarly in dispute. Although controlled ovarian hyperstimulation with both human menopausal gonadotropins alone and in conjunction with GnRH agonists have produced pregnancies when coupled with intrauterine insemination, it was demonstrated that there was a significantly greater pregnancy rate per cycle with the use of a GnRH agonist in a recalcitrant infertile population. Others did not substantiate this improvement in pregnancy rate per cycle in their patient population of regularly ovulating women undergoing their first controlled ovarian stimulation cycle either with or without GnRH agonist therapy. This suggests that women with ovulatory dysfunction, and particularly women who previously have not responded to therapy with human menopausal gonadotropin therapy, will reap the most benefits from the addition of a GnRH agonist to their ovulation induction regimen. The addition of a GnRH agonist to controlled ovarian hyperstimulation is a highly effective method of inducing pregnancy in a recalcitrant infertile population. Patients who did not conceive with human menopausal gonadotropins-intrauterine insemination may conceive with GnRH agonist-human menopausal gonadotropins-intrauterine insemination therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403618 TI - Gonadotropin-releasing hormone and its analogues: applications in gynecology. AB - In ovulation induction with hMG for IVF, clinical advantage has been taken of the agonistic ability of GnRH-a and its hypothalamic downregulating effect. This allows a better follicular synchronization, which represents an increase in the number and quality of oocytes and a decrease in the number of cancelled IVF cycles. Furthermore, according to the E2 response during the first 5 days of stimulation, the leuprolide screening test allows us to individualize ovarian stimulation for IVF and helps to prevent the potential risk of multiple pregnancy. In our experience, the majority of patients undergoing IVF benefit from the flare-up protocol, with the exception of the patients with an E2 pattern C response who benefit from the LPL protocol. PMID- 8403619 TI - Pulsatile gonadotropin-releasing hormone therapy for ovulatory disorders. AB - Pulsatile GnRH remains a physiologic method of inducing ovulation that is effective and safer than other parenteral preparations. Its lower rate of acceptance in the United States stands in curious contrast to its widespread usage in other countries as a second-line (postclomiphene) technique of choice for ovulation induction. In a high-technology era such as ours, women who may benefit from pulsatile GnRH therapy should not be forgotten. By far the most favorable results are obtained in women with primary or secondary hypothalamic amenorrhea. In such women, pregnancy rates appear comparable to those achieved with exogenous gonadotropins with a much lower risk of multiple pregnancy and ovarian hyperstimulation. These positive aspects, combined with the decreased need for clinical monitoring and the increased sense of control imparted to the patient, lead to the conclusion that women with uncomplicated hypothalamic pituitary disorders are the ideal patient group to consider for therapy. The application of pulsatile GnRH therapy to other groups of women relies on limited data. By all means, women with polycystic ovarian syndrome who have not ovulated or conceived after other forms of treatment should be considered because reasonable pregnancy salvage can be obtained. We have noted a first-cycle successful pregnancy after failure of exogenous gonadotropins, with or without a superimposed GnRH agonist, and even in vitro fertilization with multiple embryo transfers. Before such invasive, high-technology procedures are entertained, it would appear prudent to consider this simple alternative in women with an overactive hypothalamic-pituitary axis, as seen in polycystic ovarian syndrome. Other ovulatory defects also may be amenable to treatment with pulsatile GnRH, but their practical usefulness will await further clinical study. PMID- 8403620 TI - Combined use of gonadotropin-releasing hormone and its analogues for ovulation induction optimization. PMID- 8403621 TI - Gonadotropin-releasing hormone antagonists: effects on the ovarian follicle and corpus luteum. PMID- 8403622 TI - Applications of gonadotropin-releasing hormone analogues in the treatment of premenstrual syndrome. AB - A regimen in which a GnRH agonist is used with estrogen-progestin replacement provides a significant potential advance in the treatment of PMS. Substantial evidence indicates that the pathophysiology of PMS is dependent on cyclic progesterone changes which, after a time delay, influence central neurotransmitter systems and peripheral tissues. Because of the myriad neurotransmitter changes induced by progesterone, the most comprehensive treatment of the complex array of symptoms in severe PMS may require pharmacologic agents that reduce circulating progesterone levels. The most effective agent in this regard in a GnRH analogue. Until recently, use of these analogues has been limited to short-term courses as a result of the deleterious effects of hypoestrogenism. Based on newer information, a regimen in which a GnRH agonist is used concomitantly with CEE and MPA replacement appears to ameliorate PMS symptoms substantially while providing a proven estrogen-progestin replacement therapy to protect against the side effects of hypoestrogenism. Before treatment, an accurate diagnosis must be made that rests on exclusion of concomitant psychiatric and/or medical diagnoses and prospective symptom recording. In addition, before use of the GnRH agonist and estrogen-progestin add back treatment, consideration should be given to agents, such as fluoxetine and alprazolam, which target specific neurotransmitter alterations. Currently, the superiority of one agent over others has not been tested in controlled trials. Nonetheless, in patients who cannot tolerate or do not respond to fluoxetine and alprazolam, a GnRH agonist plus estrogen and progestin appears the indicated treatment of choice. PMID- 8403623 TI - Gonadotropin-releasing hormone agonists in the treatment of girls with central precocious puberty. AB - The onset of puberty before the age of 8 years in a girl is considered precocious. A child who presents with premature sexual development requires a thorough history, physical examination, and appropriate laboratory evaluation. Making the correct diagnosis is crucial to the selection of the appropriate form of therapy and management. Generally, CPP is the result of premature activation of the hypothalamic-pituitary-gonadal axis and can be successfully managed with long-acting GnRH agonists. In addition, GnRH analogue therapy has been shown to be safe, effective, and reversible. Treatment has resulted in a delay in the progression of secondary sexual development, normalization of the growth velocity, slowing of the rate of bone maturation, and an increase in the predicted final adult height. The GnRH agonists are ineffective in the therapy of gonadotropin-independent precocious puberty. PMID- 8403624 TI - Potential of gonadotropin-releasing hormone agonists in the diagnosis of pubertal disorders in girls. PMID- 8403625 TI - Focal brain invasion as the first manifestation of Langerhans cell histiocytosis in an adult. Case report. AB - A 31-year-old overweight man, suffering from high-blood pressure, was hospitalized for transient fits and hemiparesis. MRI disclosed a large irregular mass affecting the vault, meninges and invading the parietal lobe. At neurosurgery, the lesion was necrotic, hemorrhagic and poorly demarcated from the surrounding brain. Histopathology revealed a benign Langerhans cell histiocytosis. No other systemic nor organic lesions could be discovered. After additional local radiotherapy, the patient recovered completely and regained normal activities 6 months later. PMID- 8403626 TI - Hypoxia-ischemia and thiamine deficiency. AB - In order to test the hypothesis that Wernicke's encephalopathy is of topographic rather than of pathogenetic specificity we examined the brains of 49 patients without any evidence of chronic alcoholism. They had died at least four days after an event of severe hypoxia-ischemia. They all showed extensive lesions in the cortex, in the thalamus and in other regions. In 19 of them there was additional necrosis in the mamillary bodies which apparently was of the same age as the associated cortical and thalamic lesions and which could not be distinguished from Wernicke's encephalopathy. In three of the 19 cases there was a total necrosis within the mamillary bodies. By re-examining the mamillary bodies of 12 known alcoholics without any evidence for an ischemic impact we could affirm that total necrosis may fit into the spectrum of Wernicke's encephalopathy. Our findings demonstrate that the morphological changes in the mamillary bodies due to thiamine deficiency and those due to hypoxia-ischemia may be identical. PMID- 8403627 TI - Multiple sclerosis with bilateral continuous cystic lesions along lateral ventricles and caudate-callosal angles (Wetterwinkel). AB - We report a 53-year-old female autopsy case of multiple sclerosis with bilateral continuous cystic lesions along the lateral ventricles and caudate-callosal angles (Wetterwinkel). The pathophysiological mechanisms underlying these peculiar huge cystic lesions can be explained by the appearance of necrotic tissue during the recurrent relapsing stages of the disease, and then, by the absorption and scavenging of activated microglias. Poor astrocytic gliosis, which might be an effect of frequent use of corticosteroids during the clinical course makes the cavities bigger. PMID- 8403628 TI - GM2D gangliosidosis B1 variant in a boy of German/Hungarian descent. AB - After the introduction of 4-methylumbelliferyl-2-acetamido-2-deoxy-beta A-D glucopyranoside (4MUG) and its sulfated form (4MUGS) in the pre- and postnatal diagnosis and carrier identification of gangliosidosis genotypes, infrequent forms of the GM2 gangliosidosis Type B (Tay-Sachs disease) have been observed which show normal activity of Hexosaminidase A (Hex A) isoenzyme with the substrate 4MUG but absent or deficient activity against the sulfated form 4MUGS. Here we report the observation of a German/Hungarian boy aged 12 when he died with a prolonged course of a neurodegenerative disorder, later biochemically identified as a GM2 gangliosidosis B1-variant which is characterized by a deficient Hex A activity only against 4MUGS. The first clinical symptoms had occurred after the age of 14 months with a clear manifestation of the disease at age 3, when he presented disturbances of movement and tended to fall down. The slowly progressive course with brain atrophy, seizures and severe mental deterioration resulted in death after almost 9 years. At autopsy, the typical light microscopic neuronal changes of a "lysosomal storage disorder" were found, with multilamellar concentric bodies (MCB) and Zebra bodies in the neuronal cytoplasm at the electron microscopic level. PMID- 8403629 TI - Eosinophilia-myalgia syndrome: a clinicopathological study of four patients. AB - Eosinophilia-myalgia syndrome is an idiopathic disorder characterized by myalgia, especially of the extremities and peripheral eosinophilia. It is found in some patients as a result of tryptophan ingestion. We examined four patients showing clinical features and muscle biopsy changes consistent with eosinophilia-myalgia syndrome. The results of conventional histology were compared with immunohistochemical studies using monoclonal antibodies to human T lymphocytes, macrophages, HLA-DR antigen. Our findings confirm the potential importance of lymphocytes and macrophages in this syndrome. In particular, new observations are presented concerning the immunoreactivity of HLA-DR antigen. PMID- 8403630 TI - Multiple primary intracranial tumors and association of intra- and extracranial tumors. An autopsy study. AB - In 37504 autopsies performed during the last 3 decades at the Department of Pathology of Albert Szent-Gyorgyi Medical University (Szeged, Hungary) gliomas were found in 498 cases. These gliomas were associated with other types of intra- or extracranial tumors in 1.2-3.2% of the cases. Despite the male predominance observed in other types of multiple tumors, the multiple intracranial tumors and the intracranial tumors associated with extracranial malignancies were more frequent in females (possible role of hormonal influences). A relatively frequent association of various intracranial tumors with gastrointestinal carcinomas and in one quarter of the cases with extracranial benign tumors indicates that genetic factors may be involved in the pathomechanism. PMID- 8403631 TI - Skeletal muscle in children with congenital myotonic dystrophy in the first year of life. AB - Muscle biopsies of 9 children with congenital myotonic dystrophy were taken during the first year of life. All but one biopsy were abnormal showing nonspecific changes like an increased variability of fiber size, a selective atrophy of type I or type II fibers or an increased number of intrafusal muscle fibers. Nevertheless correct diagnosis of the disease based on morphological data only is impossible in some cases. A maturational arrest of the muscle is not a obligate feature of the disease. We conclude that muscle biopsy should be postponed after the first year of life. PMID- 8403632 TI - Adult-onset motor neuron disease with basophilic intraneuronal inclusion bodies. AB - A 53-year-old man initially displayed muscle atrophy and weakness and a slight degree of pyramidal signs, which became more prominent with time. Later, gaze palsy, autonomic dysfunction, and bedsore developed. The total clinical course was 63 months. Pathologically, there was loss of upper and lower motor neurons and other regions, as well as degeneration of the anterior and lateral funiculi of the spinal cord. Round or irregularly-shaped basophilic inclusions were found not only in the motor neurons, but also in other regions such as the putamen, globus pallidus, thalamus, subthalamic nucleus, red nucleus, midbrain tegmentum, pontine nucleus, dentate nucleus, inferior olivary nucleus and others. The inclusions consisted of randomly arranged 12-25 nm thick filaments studded with granules. This patient with adult onset of the disease exhibited features resembling those of "juvenile amyotrophic lateral sclerosis" with basophilic inclusions. Clinically as well as pathologically, the disease manifestations involved many systems other than the motor neurons. PMID- 8403633 TI - The role of local antibiotic therapy in the management of compound fractures. AB - Seven hundred four compound fractures (198 [28%] Grade I, 259 [37%] Grade II, and 247 [35%] Grade III) were treated during a seven-year period at the author's institution. One hundred fifty-seven open fractures (22%) (Group A) received systemic antibiotic prophylaxis only, whereas 547 compound fractures (78%) (Group B) were treated with local application of antibiotic beads (tobramycin) in addition to prophylaxis. Fracture grades, age, gender, fracture location, and length of follow-up period were not significantly different between the two groups. All fractures underwent timely irrigation, debridement, and skeletal stabilization. Forty-nine of 704 compound fractures (7%) developed an infection (acute wound infection or chronic osteomyelitis or both). Group A showed an infection rate of 17% (26/157); treatment in Group B resulted in 23 compound fracture infections (4.2%). The difference in the incidence of infection was statistically significant. Comparison of the infection rates in either wound infection or chronic osteomyelitis showed a trend toward decreased rates in Group B versus Group A throughout all fracture grades. However, by subdivision into the fracture grades, only the IIIB types had a statistically significant decrease of infection in Group B versus Group A; the wound infection rate was 39% (9/23) in Group A and 7.3% (7/96) in Group B. The rate of chronic osteomyelitis was 26% (6/23) in Group A and 6.3% (6/96) in Group B. Prophylactic use of antibiotic laden PMMA beads in addition to systemic antibiotics was of benefit in preventing infectious complications in compound fractures, in particular in Type IIIB open fractures. PMID- 8403634 TI - Septopal beads and autogenous bone grafting for bone defects in patients with chronic osteomyelitis. AB - Thirty-five patients with chronic osteomyelitis were treated with autogenous bone grafts for bone defects. Of 35 patients, two were lost to follow-up evaluation. There were 27 men and six women. The age range was from 18 to 62 years (median age, 29 years). The bones included 22 tibias, nine femurs, and two ulnas. The size of the bone defects ranged from 2.5 x 15 cm (median defect, 2.5 x 6 cm). The length of infection ranged from two to 540 months (median, 31 months). Twenty patients had nonunions and 19 patients required soft-tissue muscle transfers. Patients were treated with systemic antibiotics and/or gentamicin (Septopal) antibiotic beads. Twelve patients were treated with Septopal beads. The range of follow-up evaluation was 24 to 68 months (median, 47 months). Thirty-one of 33 patients had one bone-graft procedure and two patients required two. The time from initial debridement and infection control to bone graft for patients not requiring soft-tissue muscle flaps was one to six weeks (median time, four weeks). For patients requiring soft-tissue muscle transfer, the range was six to nine weeks (median, six weeks). The median time to bone-graft incorporation was six months. All fractures united. Complications included two refractures from auto accidents in the immediate postoperative period, one partial muscle loss, one skin-graft loss, four pin tract draining sites, and two antibiotic-related skin rashes. There was one recurrent infection, and the infection arrest rate was 97%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403635 TI - Mortality after elective total hip arthroplasty in elderly Americans. Age, gender, and indication for surgery predict survival. AB - Medicare Part A claims records were used to determine the perioperative mortality and cumulative postoperative survival of elderly Americans undergoing total hip arthroplasty (THA) for reasons other than hip fracture. The indication for THA was determined by analysis of the diagnosis codes in the claims records. Perioperative mortality was 0.95% (48/5078) and increased with age, from 0.34% (three of 872) in those 66 years to 69 years of age to 3.75% (nine of 240) among those 85 years of age or older. Perioperative mortality was not related to race, gender, or the indication for which the surgery was performed. Among these elderly patients not operated on for hip fracture, cumulative survival after surgery increased with decreasing age, female gender, and when osteoarthrosis was the indication for surgery. Persons undergoing THA had better survival after surgery than the age-gender-matched general population. Perioperative mortality among elderly Americans undergoing THA in the community is low and similar to that estimated by the 1982 NIH Consensus Conference, which considered primarily data reported from referral centers. PMID- 8403636 TI - Total hip arthroplasty in the neglected congenital dislocation of the hip. A five to 14-year follow-up study. AB - A retrospective study was performed on 34 hips of 28 patients with neglected congenital dislocation of the hip after cemented total hip arthroplasty (THA) using a straight-stem prosthesis in 21 and a curve-stem prosthesis in 13 hips. The mean age at operation was 49 years (28-72 years) and the mean follow-up period was 9.4 years (5.6-14 years). Functional evaluation using Merle d'Aubigne Hip Score showed 71% success rate. Radiographic loosening occurred in three femoral and eight acetabular components attributable to poor cementing in all stems and three sockets, and to dislocation due to trauma and graft collapse in two sockets. Marked acetabular deficiency showed increased incidence of socket loosening. Both types of prostheses showed the same statistical results. Localized endosteal osteolysis appeared in five hips at the sites of less than 1 mm cement thickness. Greater trochanter nonunion reduced the functional status evidenced by pain, limping, and a statistically significant Trendelenburg sign. Proper cementing, effective acetabular reconstruction, and prevention of trochanter nonunion can produce excellent long-term results. PMID- 8403637 TI - The Ilizarov fixator and polymethylmethacrylate-antibiotic beads for the treatment of infected deformities. AB - Eighteen of the 124 patients enrolled in the gentamicin methylmethacrylate bead (GMB; Septopal) study had infected deformities that were treated with the Ilizarov external fixator. Nine of these patients (Group 1) were treated with an initial debridement and intravenous antibiotics for four weeks. Nine patients (Group 2) were treated with GMB and, five days after operation, intravenous antibiotics. The two groups were similar. Group 1 included eight men and one woman (average age, 31.8 years) and Group 2, seven men and two woman (average age, 37.4 years). Both groups had multiple deformities, including nonunions, segmental defects, angulation, and shortness. The end results were identical. Eight patients in each group had their infection successfully arrested (88.9%), and eight patients in each group had their deformity successfully treated. The use of GMB with the Ilizarov fixator was as effective as long-term parenteral antibiotics with the Ilizarov fixator for the treatment of infected deformities. PMID- 8403638 TI - Translateral surgical approach to the hip. The abductor muscle "split". AB - A modified translateral approach to the hip, "the abductor muscle split", has been used when performing primary total hip arthroplasty. The results of this surgical approach in 50 patients with an average follow-up period of 2.8 years showed excellent restoration of abductor muscle function and gait without compromising surgical exposure. There was one case of postoperative dislocation. Heterotopic bone formation was not a clinical problem. PMID- 8403639 TI - Incidental metastatic mammary carcinoma in a total knee arthroplasty patient. AB - Incidental metastatic breast carcinoma was discovered by histologic examination in a total knee arthroplasty (TKA) patient. A 70-year-old woman was evaluated for increasing debilitating pain in the right knee. A radiograph demonstrated only degenerative changes. Degenerative changes were noted in the gross and microscopic examination of the knee specimen. The unsuspected focus of poorly differentiated adenocarcinoma was suggestive of mammary carcinoma. A mass in the right breast then was subsequently noted on reexamination. Mastectomy showed infiltrating adenocarcinoma with metastases to axillary lymph nodes, and diffuse involvement of the skeleton was demonstrated with a bone scan. This case emphasizes the importance of a meticulous history and a complete medical examination before surgery, detailed examinations of the excised knee and joint fragments by a pathologist after a TKA operation. Malignancy can be missed with only gross specimen evaluations and radiographs. PMID- 8403640 TI - Early migration of uncemented porous coated anatomic femoral component related to aseptic loosening. AB - Primary total hip arthroplasties were performed in 108 patients (116 hips) using a porous coated anatomic (PCA) hip system. Nineteen (22 hips) of 108 patients (116 hips) who had a subsidence of a PCA femoral stem more than 3 mm were studied for the time onset, the amount of stem subsidence, and outcomes. A minimum follow up period was seven years. The average age of the patients at operation was 37.4 years. The operative diagnosis was avascular necrosis of femoral head in seven hips; osteoarthrosis secondary to childhood septic hip joint in six; fracture of femoral neck in three; and miscellaneous in six hips. The average preoperative Harris hip score was 57 points, which improved to 82 points at the seven-year follow-up examination. Overall, there was a 79% prevalence of thigh pain (15 patients). Six patients had occasional, nondisabling, activity-related thigh pain. The remaining nine patients had persistent and disabling thigh pain at the final follow-up examination. All stems had an undersized component in one or both planes. One hip was stabilized by bone ingrowth. Twelve hips were fixed by fibrous tissue ingrowth and had occasional clinical symptoms. Nine hips had component loosening at the final follow-up examination, of which four were revised and five were pending revision. Early instability of the femoral component measured by radiographs was correlated with and predictive of late radiographic loosening of the femoral component. PMID- 8403641 TI - Fracture of the femoral neck in younger adults. A new method of treatment for delayed and nonunions. AB - Femoral neck fractures in young patients present a special problem because ischemic necrosis of the femoral head frequently follows. Total hip arthroplasty is not recommended because of the young age, but other methods of treatment are controversial. The best approach is prevention of the development of ischemic necrosis when treatment is delayed or has failed. Although introduction of blood supply into the ischemic femoral head has been attempted using muscle pedicled bone struts, the results have not been encouraging. A new technique using a vascularized pedicled bone strut across the fracture site, on top of the usual screw fixation, was applied to 15 cases. Fracture union occurred in all cases. After a five- to seven-year follow-up time, technical failure occurred in one case and ischemic necrosis in another. Otherwise, functional and radiographic results were satisfactory. In high-risk cases of fracture neck of femur among young adults, screw fixation combined with vascularized bone strut bridging is the recommended treatment. PMID- 8403642 TI - Complications of proximal femoral allografts in revision total hip arthroplasty. AB - Four revision hip arthroplasties using a well-described surgical construct involving a large frozen femoral allograft to replace a deficient proximal femur developed six major complications related to the use of the allograft or of the surgical construct. These cases represent three consecutive cases using this surgical construct performed by the senior author and one postoperative referral that used the same construct. The complications included three allograft fractures, two nonunions at the host-allograft junction, and one postoperative infection. All four of the patients previously had cemented revisions before the allograft procedure, and all had deficient proximal femurs at the time of the allograft procedure. All three of the fractures occurred at the tip of the femoral prosthesis within the allograft. The unacceptably high fracture rate, three of four, was associated with the allograft functioning as an unsupported load-bearing element in this study. PMID- 8403643 TI - Osteonecrosis after renal transplantation in children. AB - From January 1975 to December 1990, 141 renal transplants were performed in 129 children. There were 72 boys and 57 girls, aged two to 17 years. Osteonecrosis occurred in seven patients, in the following sites: the femoral head in four children, including two bilateral cases; and the femoral condyle in three children, with two bilateral cases. The mean period from transplantation to the diagnosis of osteonecrosis was 18 months. The osteonecrosis group was compared to the control group and no statistically significant differences were found except in the serum cholesterol levels. The corticosteroid dosage in particular, showed no correlation with the occurrence of osteonecrosis. PMID- 8403644 TI - Femoral strut allografts in cementless revision total hip arthroplasty. AB - One hundred thirteen cases of cementless femoral revision total hip arthroplasty using either cortical strut or proximal calcar allografts were reviewed for radiographic incorporation at an average 4.75 years after operation. All cortical strut grafts were used to supplement the host bone after achieving stable femoral fixation by a tight distal diaphyseal fit. Eighty-seven of the 95 cortical strut grafts showed radiographic evidence of graft incorporation. Eleven of the 18 calcar grafts had resorbed. Complications consisted of eight dislocations, three pulmonary embolisms, two superficial infections, two myocardial infarctions, and two sciatic nerve palsies. When stable distal fixation of cementless femoral prostheses have been achieved, the cortical strut allograft can be readily incorporated into the host femur and augments the host bone structure. PMID- 8403645 TI - Recurrence of pigmented villonodular synovitis of the knee 17 years after the initial treatment. A case report. AB - Pigmented villonodular synovitis (PVNS) is a benign proliferative process involving the synovial membranes bursas or tendon sheaths. The rarity of the process, the absence of well-documented long follow-up examinations, as well as reports on different forms of the disease from multiple anatomic sites has lead to some confusion. This is a case report of a 44-year-old woman with recurrence of PVNS of her right knee, 17 years after the initial limited knee synovectomy. It was proven impossible to differentiate the disease from soft-tissue malignancy despite the use of all the available investigation methods, including nuclear magnetic resonance. Eventually the diagnosis was made by an open biopsy. Marginal excision of the soft-tissue mass from the popliteal fossa was performed initially, followed by a total knee synovectomy within the next two weeks. The patient remains recurrence free 32 months after the operation. PMID- 8403646 TI - Synthetic augmented repair of proximal ruptures of the anterior cruciate ligament. Long-term results of 66 patients. AB - The long-term results (five-to eight-year follow-up evaluation) of 66 patients with high proximal ruptures of the anterior cruciate ligament (ACL) who were treated operatively are presented in a retrospective and uncontrolled study. Technique of surgery was the reinsertion of the ACL in a multiple suture loop technique, augmented with Kennedy-LAD (ligament augmentation device) on over the top route in temporary double-end fixation. This technique was used in patients with proximal rupture of the anterior cruciate ligament found at arthroscopy. In the follow-up as well as in instrumented measurement, 97% of the knee joints have normal joint laxity. According to the evaluation sheet designed by the Orthopadische Arbeitsgemeinschaft Knie (OAK), excellent or good results were found in 86% of the patients. Nine percent had limited range of motion. The ACL reconstruction technique allowed 75% of the patients to regain their preinjury sports activity level. The potential advantages of synthetic augmented reinsertion of the ACL are anatomic reconstruction without destruction of other anatomic structures as grafts; securing early rehabilitation with weight bearing of the operated limb depending on individual pain tolerance; and presenting excellent long-term results of normal joint laxity. PMID- 8403647 TI - Patellar tendon or Leeds-Keio graft in the surgical treatment of anterior cruciate ligament ruptures. Intermediate results. AB - In a prospective randomized study, 60 patients with unilateral chronic anterior cruciate ligament (ACL) rupture were allocated to surgical reconstruction using an autogenous patellar tendon graft (PT) or a synthetic Leeds-Keio graft (LK). Fifty-five patients (26 PT, 29 LK), 32 men, 23 women, fulfilled the criteria to be further tested. The mean time from surgery to follow up was 28 months. Laxity was tested by pivot shift and an instrumented anterior laxity test. Subjective knee function was classified using the Lysholm score, Tegner activity score, and IKDC grading. Muscle performance was analyzed in 49 patients (23 PT, 26 LK. There was no difference between the two groups in anthropometry, activity levels (before trauma; present; desired activity), time from trauma to surgery, or time from surgery to follow-up evaluation. Neither the concentric and eccentric knee extensor peak torque ratio nor the knee extensor endurance and the one-leg hop test differed between the two groups. However, both the pivotshift and the anterior laxity were significantly greater for the LK group. On the other hand, significantly more patients in the PT group had an extensor lag. Although the results are only intermediate, the Leeds-Keio ligament does not fulfill the requirements for a satisfactory result in ACL reconstructive surgery with regard to knee-joint stability. PMID- 8403648 TI - Popliteal pseudoaneurysm after arthroscopic meniscectomy. A report of two cases. AB - Two cases of popliteal artery pseudoaneurysm after arthroscopic meniscectomy are reported. It is a very rare complication in arthroscopic knee surgery. Bypass grafting, using autogenic saphenous vein, was necessary after resection of the aneurysm in both cases. PMID- 8403649 TI - Compartment syndrome complicating tibial tubercle avulsion. AB - Avulsion of the tibial tubercle is an uncommon physeal injury. Complications from this fracture are infrequent. Adolescent boys developed compartment syndrome after tibial tubercle avulsion. Injury to the soft tissue surrounding the tibial tubercle avulsion may be more extensive than is usually appreciated. The anatomy of the proximal tibia and the tibial tubercle with nearby branches of the anterior tibial recurrent artery suggest a predisposing factor for the development of compartment syndrome. Compartment syndrome should be added to the list of possible complications of tibial tubercle avulsion fractures. PMID- 8403650 TI - Medial unicompartmental arthroplasty. A survival study of the Oxford meniscal knee. AB - One hundred twenty-one knees with medial compartment osteoarthrosis were treated by unicompartmental arthroplasty with the Oxford Knee. The strict selection criteria were (1) the presence of a functioning anterior cruciate ligament, (2) fully correctable deformity, and (3) full thickness of articular cartilage in the lateral compartment. The mean elapsed time from surgery was 44.4 months. One knee has required revision for a loose tibial component. The results at this stage are as acceptable as those of tricompartmental knee arthroplasty and better than those of high tibial osteotomy. PMID- 8403651 TI - Pseudoaneurysm after arthroscopy of the knee. A case report. AB - Pseudoaneurysm of the inferior lateral genicular artery occurred in a 25-year-old man after arthroscopy. The arthroscopy was not complicated by postoperative bleeding or hemarthrosis, which has been associated with pseudoaneurysm formation. The diagnosis was confirmed on 99mTc radionuclide scanning and ultrasound, and the anatomy was demonstrated by digital subtraction angiography. Exploration and excision were performed without complication or recurrence. There are several methods for investigating these rare lesions. PMID- 8403652 TI - Micromotion of a noncemented tibial component with screw fixation. An in vivo roentgen stereophotogrammetric study of the Miller-Galante prosthesis. AB - Micromotion of the tibial component was studied in ten cases of successful arthroplasty for gonarthrosis using the Miller-Galante total knee prosthesis with roentgen stereophotogrammetric analysis (RSA). In six of seven cases, there was a mean of 0.6-mm migration during the first year. Most of this migration occurred as tilt and rotation. Subsidence was seldom found. Inducible displacement was observed in seven cases after one year; in six, deflections were found. In five cases, the inducible displacement was small, approximately 0.3 mm, whereas the sixth case showed inducible displacement of 1.7 mm. The micromotion found in this study was smaller than has previously been reported for other noncemented prostheses. In some of these cases, the RSA data may be compatible with mechanical coupling of the prosthesis to the bone by bony ingrowth. PMID- 8403653 TI - Complications and surgical indications in 144 cases of nonmetastatic osteosarcoma of the extremities treated with neoadjuvant chemotherapy. AB - From September 1986 to December 1989, 144 patients with osteosarcoma of the extremities were treated with combined surgery and neoadjuvant chemotherapy. The disease-free survival was 79% for good responders (necrosis greater than 90%) and 72% for poor responders (necrosis less than 90%), and the local recurrence rate was low. Improvement in long-term prognosis and the increase of limb-sparing surgery determine a higher rate of immediate and late complications. Most of the complications were observed in limb-salvage procedures; 63% of these procedures presented one or more complications. In nine rotationsplasties, there were four complications, and in 13 amputations no complications were observed. Therefore, 55% of patients were affected by surgical complications. Twenty-eight complications were considered minor (not requiring surgery), whereas 77 complications were major. Functional results, evaluated according to Enneking's new system, were higher than 50% in two thirds of the limb-salvage procedures. Complications in limb-salvage procedures are more influenced by the type of reconstruction than by the surgical procedure used. Probably the most troublesome consequence of surgical complications in osteosarcoma is the deviation or delay in administering postoperative chemotherapy, which jeopardizes survival. PMID- 8403654 TI - The treatment of infected nonunions with gentamicin-polymethylmethacrylate antibiotic beads. AB - Fifty-two patients enrolled in the Septopal study of infected nonunions were prospectively examined in a randomized, controlled, closed study. Patients were divided into two groups. Group 1 consisted of 24 patients treated with debridement and intravenous antibiotics for four weeks. Group 2 consisted of 28 patients treated with debridement, gentamicin-polymethylmethacrylate (Septopal) beads, and perioperative broad-spectrum parenteral antibiotics. Both groups were treated with similar methods for reconstruction of the nonunions. The demographics of the two groups were similar. The average patient age in Group 1 was 38.4 years, and in Group 2, 37.1 years. Group 1 included 21 men and three women and Group 2, 23 men and five women. The nonunions in both groups ranged from simple hypertrophic nonunions to atrophic unions to segmental defects. The end results were good in both groups. Twenty patients in Group 1 and 25 patients in Group 2 had their infections successfully arrested (83.3% and 89.3%, respectively). Nonunions were successfully healed in the two groups, with similar results (Group 1, 83.3%; Group 2, 85.7%). Infected nonunions responded equally well to either systemic treatment with long-term intravenous antibiotics or local treatment with gentamicin-polymethylmethacrylate beads. PMID- 8403655 TI - Thermal aspects of the use of polymethylmethacrylate in large metaphyseal defects in bone. A clinical review and laboratory study. AB - The potential necrotizing effects of the heat produced by the exothermic polymerization process has raised questions regarding the use of polymethylmethacrylate (PMMA) in orthopedic surgery. An experimental model was used to: (1) analyze the amplitude and distribution of heat in bone taken from autopsy specimens when large metaphyseal defects (simulating tumor excision) were filled with curing PMMA and (2) to observe any significant necrotizing temperatures. The experimental design included two experimental groups of five distal femora into which either a small- or large-bore defect was made in the lateral epicondylar region. These defects were filled with either one or two packs of PMMA cement, and temperature probes were used to record temperature elevations at the cement core, the bone-cement interface, and the surrounding 1-, 2-, 3-, and 5-mm bone intervals. To simulate physiologic temperature and fluid environment, the experiment was conducted in a 37 degrees normal saline bath. There is clinical evidence suggesting that the large amounts of PMMA used in tumor reconstructive surgery (often two packs or more) may lead to thermal necrosis of remaining tumor cells in the curetted cavity. This may explain, at least in part, the decrease in recurrence of giant-cell bone tumor after curettage and PMMA cementation. PMID- 8403656 TI - Metastatic bone tumors. Nonsurgical treatment. Outcome and survival. AB - Of 683 (16.1%) consecutive patients investigated from October 1984 to December 1985, 110 had various malignant tumors and secondary bone involvement. In 82 of 110 patients (74.5%), the primary tumor was located in the breast, and the percentage of clinical and radiographic objective responses (OR) of their bone lesions (42.7% and 22.8%, respectively) was higher than for cancer in other sites (clinical OR, 21.4%; radiographic OR, 15.8%). The average survival time of 82 breast cancer patients was 87.4 months, or 38 months from the diagnosis of the bone metastases. Of the 110 patients with bone metastases, 60 patients (54.5%) had secondary spinal involvement. The clinical and radiographic OR (38.4% and 30.2%) were better in the patients with vertebral metastases than in the patients with bone metastases in other sites (clinical OR, 36%; radiographic OR, 9.1%). The average survival time of the patients with vertebral involvement was 99.4 months, or 40.4 months from the time of diagnosis of the bone metastases. From the results obtained, in terms of OR and survival for patients with bone metastases, early and aggressive treatment should be considered to improve the patients' life quality. PMID- 8403657 TI - Tarsal navicular position after complete soft-tissue clubfoot release. AB - The navicular position was evaluated (according to Simons' criteria) on anteroposterior and lateral roentgenograms of 45 clubfeet that were treated with complete soft-tissue release without internal fixation. The evaluation was performed an average of 28 months after surgery. Position of the navicular correlates well with the functional rating score system. Navicular position can be viewed as an indicator for clubfoot correction. Internal fixation of the talonavicular joint was correlated with favorable correction of deformity. PMID- 8403658 TI - Biochemical markers of bone metabolism in patients with fracture of the distal forearm. AB - Biomarkers for bone turnover have established a place in the investigation and follow-up examinations of patients with osteoporosis, but has received little attention for the possibility that the fracture itself might interfere with the interpretation of the results. In this investigation of patients with fracture of the distal forearm, biomarkers for bone formation (serum osteocalcin and alkaline phosphatases) and for resorption (urinary calcium and hydroxyproline) were determined together with serum calcium and parathyroid hormone (PTH) in longitudinal and cross-sectional studies. During 16 weeks of follow-up examinations, starting on the day of the fracture, 13 patients with Colles' fracture displayed a consistent pattern with a moderate increase in serum alkaline phosphatase and osteocalcin, whereas the indices of bone resorption appeared unaffected. There was also a significant increase in the serum calcium concentration and a reciprocal decrease in serum PTH. A cross-sectional comparison between 99 patients and controls showed elevated osteocalcin levels in the patients and an inverse relationship between these levels and bone mineral density. The findings demonstrate that fracture healing should be considered in the interpretation of biomarkers in osteoporotic patients, and that among patients with a fracture of the distal forearm, those with biochemical evidence of increased bone turnover have the lowest bone mass. PMID- 8403659 TI - A new concept in fracture immobilization. The application of a pressurized brace. AB - Sixteen dogs and 14 anatomic tibiae were studied to determine the effect of a pressurized brace on a canine tibial fracture model. Bilateral tibial fractures were treated with an external fixator for 48 days. At 48 days, the pressurized brace was applied to one tibia, and a conventional cast was applied to the opposite tibia. The dogs were allowed unrestricted weight bearing until 88 days. Postmortem biomechanical studies indicated that tibiae were stronger on the side treated by the pressurized brace. This was reflected in torque values, energy, and degrees of rotation. The histomorphometry of the periosteal bone of the fracture sites disclosed a more dense periosteal callus on the side treated with a pressurized brace. In vitro mechanical studies in cadaveric tibias at 30 degrees rotation indicated that the pressurized brace provided significantly more stability. PMID- 8403660 TI - Function ten years after Colles' fracture. AB - There are no data in the literature concerning the outcome of Colles' fracture beyond six years. One hundred consecutive patients with displaced Colles' fractures were reviewed ten years after the injury. Function, radiographic anatomy, osteoarthrosis, and reflex sympathetic dystrophy (algodystrophy) were all objectively assessed. By the time of this review, 35 patients had died. Eighty-five percent of those surviving had a satisfactory outcome. Forty-two percent had improved functionally in ten years and 20% had deteriorated. Initial and ten-year radial shortening and early finger stiffness significantly correlated with final outcome. Dorsal angulation influenced early but not ten year function. Sixty-two percent of those with an unsatisfactory result had objective features of reflex sympathetic dystrophy, compared with only 6% of those with a satisfactory result. Osteoarthrosis was found in 37%, but in only 4% was it associated with an unsatisfactory outcome. PMID- 8403661 TI - Deep venous thrombosis in orthopaedic patients. Improving the specificity of diagnosis. AB - The diagnosis of deep vein thrombosis after total joint arthroplasty is difficult. The most widely used method of detection is physical examination and selective venography, which are unreliable and expensive even when symptoms and signs are prominent. This study employed computerized strain gauge plethysmography (CSGP) to select symptomatic patients for venography. The incidence of detection by CSGP was increased from 16% to 81.7%. Repeated CSGP also was investigated, but although specificity was further enhanced, it was at the expense of sensitivity. The negative predictive value of the CSGP screening was high and comparable to that of venography and indicates that CSGP is useful in the management of patients with symptoms suggestive of deep vein thrombosis. Computerized strain gauge plethysmography results emphasize the low specificity and poor feasibility of clinical examination and venography. Computerized strain gauge plethysmography is strongly advocated as a selection procedure for invasive venography in total joint arthroplasty patients. PMID- 8403662 TI - Quality of life assessment of patients with posttraumatic fracture nonunion, chronic refractory osteomyelitis, and lower-extremity amputation. AB - One hundred nine patients with long-bone fracture nonunion, chronic refractory osteomyelitis, and posttraumatic amputation were evaluated to assess the impact of chronic disability on the quality of life. The quality of life parameters were defined by a functional assessment instrument, the Arthritis Impact Measurement Scale (AIMS), and a Psychosocial Adjustment to Illness Scale (PAIS). A spouse PAIS self-report instrument was administered to assess the psychosocial adjustment of spouses or significant others. A final questionnaire was developed to determine the reasons, in order of their importance, for either continuing medical therapy or accepting amputation. The PAIS scores differed significantly between osteomyelitis patients and nonunion or amputation patients. The presence or absence of pain produced significant differences in AIMS and PAIS scores of nonunion and osteomyelitis patients. Subscale analysis of AIMS scores showed significant differences among the three groups with respect to health perception and scale of orthopaedic problem. The osteomyelitis patients were more severely affected than the nonunion or amputation patients. The PAIS was unable to detect any statistically significant differences in psychosocial adjustment of the spouses of patients in each of the three population groups. The most frequent reason for continuing medical and surgical management of nonunion and osteomyelitis was hopeful expectation for cure. The group who chose amputation did so in an attempt to put an end to the need for medical and surgical treatment. Differences in psychosocial and functional ability were related to disease diagnosis, pain, status of fracture healing, and timing of amputation. This study provides further insight into the quality of life experience for patients with long-term orthopaedic problems. PMID- 8403663 TI - Indications and results of vascularized pedicle iliac bone graft in avascular necrosis of the femoral head. AB - Several reports describe methods of treatment for avascular necrosis of the femoral head (ANFH) involving 0 to 2 mm of collapse. Some cases of ANFH have good prognoses, requiring only non-weight-bearing treatment. Other cases rapidly progress to collapse and complete destruction with enlargement of the necrotic area. The progression of the necrotic area is related to the activity of the original disease, steroid treatment, and the size and location of the necrotic area in the early stages of the disorder. In this report, a vascularized pedicle bone graft was used to treat ANFH, particularly those cases identified as Stage II on the system established by the Japanese Investigation Committee. Surgery involved curettage of necrotic bone, implantation of spongy bone, and application of a vascularized pedicle bone graft. Grafts were taken from the ilium and included the superficial circumflex iliac artery (SCIA). A bony canal was made in the anterior femoral neck, from which the necrotic bone was curetted and to which the bone graft was applied. The deep circumflex iliac artery (DCIA) was also used in combination with the SCIA. The postoperative weight-bearing period was six months. Follow-up periods lasted one to six years. Seventeen of 23 Stage II joints (19 cases) achieved satisfactory results at a mean of three years after surgery. Three Stage II joints and three Stage III joints continue to have significant problems. One of these six has been converted to a dual-bearing type endoprosthesis. The unsuccessful results generally occurred in patients who were treated with steroids. PMID- 8403664 TI - Tendon insertions onto allografts pretreated with heat and/or bone surface demineralization. AB - Tendon insertions onto allogeneic bone grafts were studied histologically and biomechanically in 132 rats. Before grafting, allogeneic bone was treated at different temperatures or partially demineralized or both. Mesenchymal cell and fibrous tissue invasion were easily recognized in non-heat-treated allograft groups and in groups with allografts incubated at 70 degrees; invasion was not observed in autoclaved (130 degrees) allograft groups. New bone formations were found between the tendon and the surface-demineralized allografts in the non-heat treated and 70 degrees-treated bone groups but not in the autoclaved group. Allografts pretreated with heat demonstrated graft-insertion tensile strengths lower than in non-heat-treated and 70 degrees-treated allografts. Surface demineralization enhanced graft-insertion tensile strengths in the non-heat treated and 70 degrees-treated groups, whereas the tensile strength of attachments in noninductive, autoclaved allografts was not enhanced. PMID- 8403665 TI - Surface adhesion and attachment factors in bone morphogenetic protein-induced chondrogenesis in vitro. AB - Bone morphogenetic protein (BMP) induces cartilage and bone development in embryonic and post-fetal life. Bone morphogenetic protein-induced chondrogenesis in outgrowths of muscle connective tissues on various furrowed and unfurrowed fibronectin-coated substrata was observed by biochemical and histologic methods. The substrata consisted of cellulose acetate membranes, atelocollagen, denatured autoclaved demineralized bone matrix, bone matrix deactivated by extraction of BMP with guanidine hydrochloride (GuHCl), and aggregates of insoluble BMP and associated noncollagenous proteins (BMP/NCP) prepared from bovine bone. The furrows, which were mechanically cut into the substrata, were intended to increase surface area and provide extra spaces for cell proliferation under compression. The extent to which extracellular attachments were required for induced cartilage development was reflected in the quantity of cartilage formed when BMP/NCP, either in insoluble or in water-soluble form, was introduced was greatest on GuHCl-extracted bone matrix. On cellulose acetate and atelocollagen, BMP-induced cartilage development was relatively scanty. A substratum of bone matrix, denatured by autoclaving at a minimum of 125 degrees, permitted cell-to cell adhesion but not cell-to-substratum attachments; the end product was loose fibrous connective tissue only. In contrast, cartilage development occurred on surfaces of undissolved particles of BMP/NCP. Water-soluble human BMP induced development of masses of amorphous cartilage. Even after it was extracted with GuHCl, rat bone matrix may have retained trace amounts of endogenous BMP. Thus, when the requirements of cells for cell-to-cell adhesion and cell-to-substratum attachment, including mechanical factors such as furrows to enlarge surface area, were met, cartilage development was a manifestation of temporal, spatial, and BMP distribution patterns. In situ hybridization and immunofluorescent microscopy with the aid of antirecombinant BMP antibiotics may provide new information about morphogenesis. PMID- 8403666 TI - Penicillin as a chemotherapeutic agent. 1940. PMID- 8403667 TI - Bovine bone morphogenetic protein-induced dentinogenesis. AB - Differentiation of odontoblasts is important for dentin formation in tooth germs and mature teeth. Although previous reports have indicated that there may be a kind of inductive agent that could induce mesenchymal cells in dental pulps to differentiate into odontoblasts, and secrete dentin matrix, the primary inductive factor of odontoblasts has not been found. Bone morphogenetic protein (BMP), which induces the formation of cartilage and bone when implanted in muscle tissue, is found in dentin matrix. The relationship between the differentiation of odontoblasts and BMP was observed by means of immunohistochemical staining with monoclonal antibody (MAb) against BMP in dental pulp tissue and cell culture; [3H]thymidine incorporation; and measurement of alkaline phosphatase activity. The conclusions are: (1) BMP exists in odontoblasts, ameloblasts, and dentin matrix (the positive reaction in ameloblasts appeared earlier and remained stronger); (2) BMP promotes incorporation of [3H]thymidine and increases the activity of alkaline phosphatase in cultured dental pulp cells; (3) BMP-induced dental pulp cells in dental pulp tissue cultures differentiate from mesenchymal to odontoblast-like cells; and (4) BMP induces formation of osteodentin and tubular dentin when used as a dental capping agent of dogs' teeth. Bone morphogenetic protein plays an important role in differentiation of odontoblasts and might be one of the inductive agents of odontoblasts. Further investigations of BMP as a biologic dental capping agent are warranted. PMID- 8403668 TI - Gentamicin-impregnated polymethylmethacrylate beads compared with systemic antibiotic therapy in the treatment of chronic osteomyelitis. AB - A model of chronic osteomyelitis was used to evaluate the efficacy of treatment with debridement alone; debridement plus gentamicin-polymethylmethacrylate (PMMA) bead implantation; debridement plus systemic antibiotic therapy; and debridement plus systemic antibiotics and bead implantation. Debridement with the implantation of gentamicin-PMMA beads and debridement followed by systemic antibiotics were significantly more successful forms of treatment for chronic osteomyelitis than debridement alone or debridement with the implantation of PMMA beads not impregnated with antibiotics. Debridement followed by the implantation of PMMA-gentamicin beads and the use of systemic antibiotics produced the greatest success rate. Treatment with a combination of gentamicin-PMMA beads and systemic antibiotics resulted in a 100% success rate, which, although not statistically better than either treatment alone, suggests a trend. PMID- 8403669 TI - Audiometric thresholds in osteomyelitis patients treated with gentamicin impregnated methylmethacrylate beads (Septopal). AB - Use of gentamicin-impregnated methylmethacrylate beads is a relatively new and effective method of treating osteomyelitis. Although previous studies have concluded that such therapy is associated with low systemic toxicity, no data are available relating the use of this treatment to ototoxicity and hearing loss. Audiometric thresholds from 28 cases of osteomyelitis treated by surgical debridement and long-term systemic antibiotics alone (14 subjects) or gentamicin impregnated methylmethacrylate beads either alone or combined with systemic antibiotics (14 subjects) were analyzed. Results showed one permanent threshold shift in the gentamicin-impregnated methylmethacrylate beads group (8%) versus four permanent threshold shifts in the systemic antibiotics alone group (29%). Osteomyelitis patients treated with gentamicin-impregnated methylmethacrylate beads are at no more risk and are probably at less risk of experiencing ototoxicity than patients treated with conventional long-term systemic antibiotics. PMID- 8403670 TI - Long-term implantation of gentamicin-polymethylmethacrylate antibiotic beads. AB - Fifty-two chronic osteomyelitis patients treated with gentamicin polymethylmethacrylate (Septopal) antibiotic bead chains were observed retrospectively to explore the relationship between the duration of bead implantation and clinical results. Group A contained 35 asymptomatic patients who had elective bead removal based on protocol, surgeon's preference, and patient variables. Group B consisted of 17 patients who retained the antibiotic beads. The presence or absence of redness, abscess, drainage, pain, or swelling at the wound site was documented for clinical evaluation. Findings indicate that Septopal bead chain implants exhibit higher success (remission) rates with increasing length of duration without increasing the risk of complications. PMID- 8403671 TI - The antibiotic bead pouch technique. The management of severe compound fractures. AB - In a consecutive series of 704 compound fractures, 227 open fractures in 204 patients were managed with the antibiotic bead pouch technique. There were 16 Grade I compound fractures, 83 Grade II fractures, and 128 Grade III open fractures. The mean patient age was 35.25 years (range, 14-87). The Injury Severity Score (ISS) ranged from 9 to 57, with a mean of 15. Porous plastic film is placed over the soft-tissue defect to establish a "closed" bead-hematoma fracture environment containing high local levels of antibiotic at the fracture site. All patients had serial wound debridements and parenteral systemic antibiotics (cefazolin, tobramycin, penicillin). Bead pouch changes ranged from one to seven per patient (mean, two). During these changes, 1248 cultures were taken, 78 (6.25%) of which were positive in 34 patients. Seventeen patients developed clinical signs consistent with an infection. The wound infection rate was 0% in Grade I open fractures, 1.2% in Grade II compound fractures, and 8.6% in Grade III open fractures. The osteomyelitis rate was 0% in Grade I compound fractures, 2.4% in Grade II open fractures, and 5.5% in Grade III compound fractures. Wound closure was obtained in 134 fractures with delayed primary closure of the skin, in 53 fractures with flap coverage, and in 23 fractures with split-thickness skin graft. Coverage was not completed in 17 wounds, at which time an amputation was performed or death occurred. Time of closure ranged from one to 32 days (mean, 7.1 days).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403672 TI - Antibiotic bead chains. AB - Gentamicin-PMMA chains are an effective drug delivery system for local antibiotic therapy in bone and soft-tissue infections. The gentamicin concentrations at the site of infection are far higher than after systemic application of the same antibiotic and far above the minimal inhibitory concentrations of most common pathogens. Because of the very low concentrations in the serum and urine after implantation of the antibiotic bead chains, toxic side effects are not to be feared. Radical debridement with removal of all sequestrated bone fragments and removal of all alloplastic implants is mandatory before implantation of gentamicin-PMMA chains into the infected bone cavity. Primary wound closure is necessary to achieve high local concentrations. The chains can be used for temporary or permanent filling of osteomyelitic cavities. Guidelines are given for the clinical application in sequestrating osteomyelitis, infected osteosynthesis, and infected pseudarthrosis. There are distinct advantages of this new form of antibiotic therapy in chronic osteomyelitis, such as increased patient comfort by primary wound closure, no need for prolonged systemic antibiotic therapy with toxic side effects, no irrigation-suction-drainage, early ambulation, shortening of hospitalization, and reduced cost. PMID- 8403673 TI - Diagnoses and staging. Osteomyelitis and prosthetic joint infections. AB - The diagnoses of osteomyelitis and prosthetic joint infections are usually made on the basis of clinical, laboratory, and radiographic examination. The diagnostic studies presently employed to diagnosis and assess osteomyelitis and prosthetic joint infections are described. A universally applied classification system for stratifying osteomyelitis and prosthetic joint infection would provide a framework for the evaluation of medical and surgical treatment efficacy. Such a system would enable treatment results to be compared among institutions. Staging systems currently being used are described. PMID- 8403674 TI - Comparison of the clinical efficacy and tolerance of gentamicin PMMA beads on surgical wire versus combined and systemic therapy for osteomyelitis. AB - These data do not prove a statistical superiority of conventional antibiotics or Septopal in the treatment of chronic osteomyelitis. This result, however, is complicated by the biased data set represented by the combined treatment. The data do suggest that cost of treatment is considerably less in patients who are treated with local antibiotics (i.e., Septopal) alone. The rate of adverse experiences was directly related to the use of parenteral antibiotics, with higher rates of adverse experiences in the conventional and combined treatment groups. Furthermore, the Cierny-Mader Physiologic Class had the best correlation with outcome, suggesting that host factors are probably of critical importance in inducing remission of chronic osteomyelitis. This protocol was not designed to test the role of debridement in the treatment of osteomyelitis: it was assumed that debridement would be the same in both groups. It is the investigators' strong opinion, however, that adequacy of debridement was an important determinant in quiescence or recurrence in the study patients. Similarly, there was no strict control for adequacy of soft-tissue coverage provided by local or distant tissue transfer. Again, the investigators believe that adequacy, including viability and durability, of soft-tissue covering was an important determinant for the end result in these patients. Other covariants such as smoking, history, nutritional status, and other measures of general health will be added to this model when data are available. This analysis will allow definition of the appropriate clinical situations in which use of Septopal alone or combined with parenteral antibiotic is indicated. PMID- 8403675 TI - Antimicrobial treatment of osteomyelitis. AB - Parenteral antibiotic therapy remains the mainstay of antimicrobial therapy for osteomyelitis. In acute cases, empirical treatment may be used initially, but antibiotic selection should ideally be based on deep aspiration or bone cultures with corresponding in vitro sensitivity testing. The drug exhibiting the highest bacteriocidal activity with the least toxicity and lowest cost should be chosen. Antibiotic treatment will not substitute for surgical debridement of infected devitalized bone. The length of treatment with parenteral therapy remains controversial. The six-week benchmark, which was determined largely by experience with childhood hematogenous osteomyelitis, may not be applicable to contiguous focus osteomyelitis after trauma in adults. The goal of surgical treatment is to convert an infection with dead bone to a situation with well-vascularized tissues that are readily penetrated by blood-borne antibiotics, making prolonged drug treatment unnecessary. PMID- 8403676 TI - A comparison of gentamicin-impregnated polymethylmethacrylate bead implantation to conventional parenteral antibiotic therapy in infected total hip and knee arthroplasty. AB - A multicenter study of infected total knee and total hip arthroplasties was conducted from 1985 until 1990. Twenty-eight patients (22 total hip arthroplasties and six total knee arthroplasties) who had periprosthetic infections were treated according to a prospective, randomized protocol. After initial debridement for their infections, patients were randomized into one of the two following groups: Group I, debridement and the implantation of gentamicin polymethylmethacrylate (PMMA) beads; and Group 2, debridement and conventional parenteral systemic antibiotic therapy. After initial treatment, the patients were then scheduled for a delayed reconstruction total joint arthroplasty. Of the 28 patients, 25 subsequently had delayed total arthroplasty, and if acrylic bone cement was used for fixation at the time of reimplantation, antibiotics were not added to the cement. The average follow-up period was three years (range, six months to 5.6 years). Infection recurred in two patients treated by debridement and the implantation of gentamicin-PMMA beads (15%) and in four patients treated with debridement and conventional systemic antibiotic therapy (30%). All recurrences occurred in patients who had infected total hip arthroplasties; none occurred in patients with total knee arthroplasties. The conditions that were common in patients with recurrent infection were (1) multiple previous surgeries, (2) host compromise and malnutrition, (3) extensive infection, and (4) inadequate debridement. The recurrence of infection was not statistically significantly more common in either treatment group. The outcome of treatment in patients with infected total joint arthroplasties using debridement, gentamicin-PMMA bead implantation,and a two-stage delayed reconstruction was similar to that of patients treated with debridement combined with conventional parenteral systemic arthroplasty and two-stage reconstruction. PMID- 8403677 TI - [Effects of calcitonin gene-related peptide on muscle cell differentiation and development]. AB - We studied the factors which facilitate muscle cell differentiation and development to produce preventive and therapeutic treatment for muscle atrophies. Calcitonin gene-related peptide (CGRP) is present in the nervous system and neuromuscular junction, and acts as either a neurotransmitter or a neuromodulator. In this study, we report the effects of CGRP on the differentiation and development of rat skeletal myoblasts. Rat skeletal myoblasts (L6 cell) were cultured in Dulbecco's modified Eagle's medium containing 2% horse serum and 6 micrograms/ml insulin (control group). CGRP was then added at concentrations of 1, 10, and 100 nM (CGRP groups). Changes in cell morphology, myoglobin (Mb) content, creatine kinase (CK) activity, DNA content, and calcium concentration in the cultured cells were examined on days 3, 6, 8, and 10. In all of the groups, the number of cultured cells increased until myotubes appeared on day 6. The number of myotubes then rapidly increased until day 10. Additional cell culture became impossible because the cells were detached from the culture dish. The short axis length of the cultured cells reached a maximum of 59.4 +/- 5.2 microns on day 8 in the CGRP (10 nM) group. This value was significantly greater than that in the control group (43.7 +/- 3.9 microns). The ratio of Mb content/DNA content in the cultured cells increased in all of the groups and reached a maximum of 13.6 +/- 0.4 x 10(-2) ng.Mb/microgram.DNA in the CGRP (10 nM) group on day 10. This value was approximately 3 times higher than that in the control group (4.7 +/- 0.3 x 10(-2) ng.Mb/microgram.DNA).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403678 TI - [Sequential studies of motor nerve conduction in relation to the disease stages of Guillain-Barre syndrome]. AB - We retrospectively studied sequential changes in motor nerve conduction in forearm segments of 17 median nerves in 9 consecutive patients with Guillain Barre syndrome (GBS). The disease course was divided into 6 stages: progressing, nadir, early-, mid- and late-convalescent, and cured stages. All patients were tested on at least 3 occasions in each different stage. Low amplitude compound muscle action potentials (CMAP) were noted in the progressing and nadir stages, suggesting the presence of conduction block. In the early- and mid-convalescent stages, on the other hand, the delays in distal motor latencies and conduction velocities as well as temporal dispersions of CMAPs were more prominent than in the progressing or nadir stage. These findings suggest that pathophysiological processes are different in the two stages of GBS: demyelination with conduction block predominated in the earlier stage and demyelination without conduction block predominated in the later stage. The former is considered to involve predominantly the relatively smaller motor nerves during the progressing and nadir stages, producing muscle weakness but not causing marked slowing of the motor nerve conduction. The latter is considered to involve predominantly the fastest motor fibers in the later days lasting into the early- and even mid convalescent stages. This process seems to produce marked conduction slowing, but does not cause prominent muscle weakness. In addition, recovery from the conduction block during the convalescent stages produces temporal dispersions of CMAPs because of the variably slowed conduction velocities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403679 TI - [Coagulation and fibrinolytic parameters in patients with various angiopathies- analysis in cerebral thrombosis, diabetic and vasculitic neuropathies]. AB - We measured the coagulation and fibrinolytic parameters in patients with various angiopathies to clarify the usefulness of the parameters for the evaluation of vascular involvement. The study included 65 patients with cerebral thrombosis at the chronic stage, 47 diabetics with neuropathy associated with diabetic microangiopathy, 15 diabetics without neuropathy, and 16 patients with vasculitic neuropathy associated with collagen diseases. Control subjects were 45 patients with other neurological disorders without symptomatic angiopathies. In cerebral thrombosis at the chronic stage, the coagulation factor VIII (FVIII), von Willebrand factor (vWF), and platelet factor IV were elevated significantly as compared with age-matched controls. Diabetics with neuropathy showed significantly elevated FVIII and vWF in comparison with diabetics without neuropathy and controls. No significant difference in parameters concerning glucose metabolism was observed between the two groups of diabetics. These findings suggest that elevated activities of the coagulation parameters reflect not merely the abnormalities in glucose metabolism but also the presence of microangiopathy which causes diabetic neuropathy. The patients with vasculitic neuropathy exhibited significantly elevated FVIII, vWF and fibrin/fibrinogen degradation products (FDP). The present study demonstrated that FVIII and vWF are useful parameters for the evaluation of the extent of vascular involvement in angiopathies of different etiologies. It is important for the treatment of patients with the angiopathies, to monitor the coagulation parameters which reflect vascular involvement in addition to parameters for the disease activity. PMID- 8403680 TI - [Aberrant expression of major histocompatibility complex class II antigen in inflammatory myopathies]. AB - Although major histocompatibility complex (MHC) class I molecules have been consistently demonstrated on muscle fibers of patients with inflammatory myopathies (IM), results concerning the expression of MHC class II molecules have been conflicting and mechanism of the expression remains to be elucidated. Expression of MHC and neural cell adhesion molecules (N-CAM) was analyzed in 38 cases of IM and in 13 cases of normal and disease controls by the immunocytochemical method. The sarcolemmaL HLA-ABC molecules were seen in all IM cases, and were visualized on the every muscle fiber in 68% of the cases. In contrast, the HLA-DR, -DP, or -DQ expression was seen on scattered fibers in 37%, 11%, 5% of IM cases, respectively. The HLA-DR reactivity was often seen on muscle fibers in the vicinity of infiltrating cells or on the perifascicular fibers. The HLA-DR positive fibers did not express N-CAM, suggesting a degenerating/regenerating process of muscle fibers was not contributory to the aberrant expression of MHC class II molecules. The perifascicular HLA-DR expression was related to the perifascicular atrophy and was more frequent in polymyositis complicated with interstitial pneumonitis. Those results suggest that increased expression of MHC class I molecules precede the aberrant expression of MHC class II molecules, and the perifascicular atrophy might reflect immunopathological processes. PMID- 8403681 TI - [A case of motor neuropathy with pyramidal sign due to prolonged administration of high dose of pancuronium bromide (Myoblock)]. AB - We reported a case of motor neuropathy with pyramidal sign following prolonged administration of a high dose of muscle relaxant, pancuronium bromide (Myoblock). A 40-year-old male was admitted to our hospital with acute episode of pancreatitis. He was treated with artificial ventilation and Myoblock to manage delirious state, disseminated intravascular coagulation and multiple organ failure. Total dose of 823 mg (24 mg/day) of Myoblock was given intravenously over 36 days. After Myoblock was discontinued, he regained his consciousness and marked muscle weakness with atrophy was noted in both limbs, more severe in distal lower limbs, without any noticeable sensory and sphincter disturbances. Motor nerve conduction studies showed normal nerve conduction velocities with markedly decreased amplitude of compound muscle action potentials. Repetitive nerve stimulation studies revealed decrement response after tetanic stimulation, which disappeared later. Needle EMG showed active denervation potentials and marked polyphasic motor unit potentials. Muscle biopsy revealed neurogenic muscle atrophy with fragmented acetylcholine esterase-positive postsynaptic sites. Sural nerve biopsy showed slight to moderate degree of axonal degeneration of myelinated fibers. Clinical, electrophysiological, and pathological studies above indicated that the main affected sites were neuromuscular junctions including the terminal twigs of motor neurones and postsynaptic membrane, and pyramidal tracts, predominant in lower limbs. About one month after the recognition of the muscle weakness, his muscle strength improved gradually, however, spasticity with hyperreflexia and pathologic reflexes of both legs were found, and became more prominent thereafter. Intensive physiotherapy and rehabilitation led improvement to the point that he became able to ambulate with walking-aids about 7 months later, but marked spasticity persisted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403682 TI - [Lateral medullary syndrome due to cavernous malformation in the brain stem]. AB - A 59-year-old female was admitted with complaints of vertigo, dysarthria and dysphagia. On neurological examination, right-sided cranial nerve signs included ptosis, Bruns's nystagmus, decreased corneal sensation, diminished facial pain and temperature sensation, decreased palatal excursion and loss of gag reflex. There was no evident motor weakness, but deep tendon reflexes were slightly exaggerated on the left extremities. Coordination testing showed right cerebellar signs. Sensory examination of the remaining parts of the body was quite normal. X ray CT scan showed multiple high density areas in the right medulla, right pons, right temporal and frontal lobes. T2 weighted MRI demonstrated these lesions as mixed signal intensity areas with marked low signal intensity rim. There were multiple black dots in the bilateral frontal and temporal lobes, cerebellar hemispheres on T2-weighted images. Carotid and vertebral angiograms showed no abnormality. This is the first report of the cavernous malformation presenting as lateral medullary syndrome. PMID- 8403683 TI - [Two siblings of type 3 GM1 gangliosidosis with different clinical features and different ages of onset]. AB - We experienced two siblings of type 3 GM1 gangliosidosis. A 33-year-old woman developed dysarthria, dysbasia and bradykinesia at around the age of 30. Her 28 year-old brother showed locomotor retardation and skeletal deformity in infancy. He lost the ability to stand walk at childhood, and developed progressive dystonia. The major neurologic manifestations were parkinsonian symptoms in the elder sister, and progressive dystonia in her brother. Both had markedly reduced beta-galactosidase activity in peripheral blood lymphocyte and were diagnosed as having type 3 GM1 gangliosidosis. Gene analysis revealed that these patients were homozygotes of the adult type mutant gene. The two siblings are unique in that the clinical manifestations and the age of onset of symptoms differed markedly between them despite the same mutant gene in both cases. PMID- 8403684 TI - [A case of Listeria rhombencephalitis with a secondary vasculitis suggested by MRI]. AB - We reported a rare case of Listeria rhombencephalitis with meningitis. A 48-year old healthy man suddenly experienced high fever and headache, then he had lower cranial nerve's palsies and mental dysfunction developed during one week period. On admission, his temperature was 38 degrees C. He was slightly delirious and euphoric. He had nuchal rigidity, mild paresthesia over his left cheek to left upper lip, a right sixth nerve palsy, dysphagia, hiccup, nasal voice and left cerebellar ataxia. His tongue deviated toward the right side on protrusion. A CSF culture grew Listeria monocytogenes. Intravenous antibiotic therapy (PIPC, minocycline hydrochloride) produced improvement in one month except for mild paresthesia and dysphagia. He almost recovered after 7 months of illness. Brain MRI on T2 weighted image demonstrated multiple small ischemic lesions in the left lateral medulla, upper pontine tegmentum in the right side, and pontine tegmentum in the left side. These lesions enhanced by Gd. were assumed to be due to the secondary vasculitis. Listeria rhombencephalitis is extremely rare in human beings. To our knowledge only thirteen cases have been reported. In seven cases, post-mortem pathological findings confirmed necrotizing angitis in brainstem. Clinical aspects of Listeria rhombencephalitis were discussed, and the entity of this disease should be considered as a treatable cause of acute progressive brainstem meningoencephalitis. PMID- 8403685 TI - [Double meningoencephalitis with herpes simplex virus and rubella virus]. AB - A 47-year-old man suffered from a headache, fever and memory disturbance. He was admitted to Morimoto Hospital. Neurological examination revealed disturbance of memory of recent events. Cranial nerves were normal. Muscle strength of the extremities was normal, except for Barre's sign of the right upper extremity. Deep tendon reflexes were exaggerated bilaterally, and extensor plantar response of the left side was elicited. Sensory examination showed no abnormality. Cerebellar sign was not recognized. Meningeal sign was slightly but clearly showed. CT scan demonstrated brain swelling at the right insular cortex region followed by severe hydrocephalus with dilatation of the lateral and the third ventricles. Cerebrospinal fluid showed high CSF pressure (250 mmH2O), pleocytosis (C.C. 359/mm3) and elevated protein level (213 mg/dl). Virological examination revealed herpes simplex virus (HSV) (CF) 32x, HSV-1 IgG (EIA) 4,050X, rubella virus IgE (EIA) 6,060X, cytomegalovirus (CMV) IgG (EIA) 1,130X in serum and HSV (CF) 1X, HSV-1 IGg (EIA) 1,430X, rubella virus IgG (EIA) 1,480X, CMV IgG (EIA) 587X in CSF. The ratio of serum/CSF of HSV and rubella virus titers by EIA methods were 2.83 and 4.10, respectively. He was treated by acyclovir 1,000 mg/day and gamma globulin, but his condition get worse acutely and died at 15th hospital days. This case was considered as a meningoencephalitis caused by simultaneous HSV and rubella virus infection. PMID- 8403686 TI - [A case of paraneoplastic autonomic and sensorimotor neuropathy with dysfunction in the afferent limb of baroreflex arc]. AB - A 65-year-old man visited our hospital with complaints of tingling sensation in the distal parts of his extremities and dysuria, which first appeared 2 months before admission. He had no abnormal findings on physical examination. Neurological examination revealed sensory impairment of glove and stocking type, mild motor weakness and muscular atrophy in the proximal parts of arms and legs, and absent tendon reflexes in knees and ankles. Fasciculation was observed on his shoulders and upper extremities, and myokymia on the abdominal wall and bilateral calves. He had hyponatremia, which was proved to be caused by SIADH. Anti acetylcholine receptor antibody, anti-GM1 ganglioside antibody and anti galactocerebroside antibody were detected in the serum. Chest X-ray showed mass shadows in the mediastinum, which were confirmed as malignant thymoma by needle biopsy. Orthostatic hypotension, neurogenic bladder and anhidrosis were observed by the autonomic function tests. Lesions responsible for orthostatic hypotension and SIADH were suspected in the afferent fibers from baroreceptors, since an reactive increase of plasma arginine vasopressin to orthostatic hypotension was blunted and reflex hypertension in the cold pressor test was well-preserved, while overshoot in Valsalva's maneuver was absent. It is important that afferent baroreceptor dysfunction may be associated with paraneoplastic neurological syndrome, since lesions in acute autonomic neuropathy are usually in the efferent fibers. PMID- 8403687 TI - [Slow eye movements (slow saccades) in Wilson's disease]. AB - Slow eye movements or slow saccades are relatively rare conditions in Wilson's disease where selective impairments of saccades are observed. The authors have a case of Wilson's disease showing a defect of saccades in all directions, with the complete preservation of smooth pursuit eye movements. T2-weighted images revealed abnormalities of signal intensity in bilateral pontine tegmentum, besides putaminal and thalamic lesions commonly seen in the previous reports. A rather selective disturbance of saccade eye movements due to the pontine tegmentum is consistent with the concept, as yet mainly based on experimental work on animals, of an anatomical segregation of the brainstem pathways for smooth pursuit eye movements, saccades, and vestibular and optokinetic patterns. Whereas the basal brainstem, especially basal pons, subserves smooth pursuit eye movements, saccades and reflex-like eye movements are mediated by the pontine tegmentum. The authors' case offers a support that dissociating eye movement disorders are due to the restricted lesions mentioned above. PMID- 8403688 TI - [A family with MELAS whose main manifestations are maternally-transmitted deafness and diabetes mellitus]. AB - A family with maternally-transmitted deafness and diabetes mellitus is described. Although the proband clinically exhibited MELAS-like symptoms such as sudden onset cerebellar ataxia and weakness of the proximal portion of the limbs in addition to deafness and diabetes mellitus, the other three members of the family had only deafness and diabetes mellitus and no neurological manifestations. The analysis of mitochondrial DNA of the two members revealed an A-->G mutation of tRNA(leu(UUR)), a mutation commonly seen in patients with MELAS. According to the clinical histories and endocrinological investigations, the type of the diabetes mellitus in this family was considered to be IDDM, which may be attributed to the dysfunction of mitochondrial of the pancreas islet cells, resulting from the mutation of the mitochondrial DNA. PMID- 8403689 TI - [Treatment of Guillain-Barre syndrome with high-dose intravenous immunoglobulins- a comparison with plasma exchange]. AB - We used high-dose intravenous immunoglobulin (IVIG) in the treatment of Guillain Barre syndrome (GBS) and compared its therapeutic effects with those who were treated by plasma exchange (PE). For this study, we selected patients who had been within seven days from the onset and unable to walk without support at that time. Four patients (3 male, 1 female, mean age 41.8) were treated by plasma exchange with 3,000-4,000 ml exchange of plasma/day for 4-6 days. Other four patients (2 male, 2 female, mean age 45.7) were treated with intravenous high dose immunoglobulin: 400 mg/kg/day for 3-5 days. The functional scale was measured at study entry and after the treatment daily. The mean times to the recovery of independent walk (a functional grade 2) were 23.8 and 10.0 days for PE and IVIG respectively. Although there was a difference of disease severity between these two groups, IVIG was at least as effective as PE in the treatment of GBS. PMID- 8403690 TI - [Prion protein coden 129 polymorphism (Val/Met) in a case of sporadic progressive multisystem degenerative disease]. PMID- 8403691 TI - Ga-67 imaging in primary and secondary psoas abscess. AB - Primary and secondary psoas abscesses are uncommon and frequently misdiagnosed. Two cases of pyogenic psoas abscess are presented to illustrate the usefulness of Ga-67 scintigraphy in determining the presence and the extent of the infectious process. PMID- 8403692 TI - Ga-67 scintigraphy in a child with adrenocortical carcinoma. AB - Adrenocortical carcinoma is a rare malignancy in childhood and often presents with a large abdominal mass and precocious puberty. The tumor is often advanced at the time of diagnosis. Radiological techniques are the main imaging modalities for assessment of the primary tumor. Ga-67 scintigraphy has been used in other pediatric solid tumors, however, no data were available for this tumor in childhood. A case of a 2.5-year-old male with a Ga-67 avid adrenocortical carcinoma is reported in which Ga-67 scintigraphy was very useful in detecting recurrence of the tumor and in assessing response to therapy. PMID- 8403693 TI - Lymphoscintigraphy. A reliable test for the diagnosis of lymphedema. AB - The efficacy of lymphoscintigraphy in the diagnosis of lymphedema was evaluated in 17 patients (total of 20 extremities). Final diagnoses were: three extremities with primary lymphedema, eight with secondary lymphedema, and nine with edema due to other causes. Qualitative interpretation of image patterns was reliable in differentiating lymphedema from edema of other origins (sensitivity 73%, specificity 100%). There were eight true-positive, nine true-negative, no false positive, and three false-negative extremities. The three false-negative results occurred in extremities that were not imaged within the first hour. Lymphoscintigraphy is a reliable, objective, and noninvasive means of supporting the diagnosis of lymphedema, and lends itself more readily to repeated studies for follow-up than other available tests. PMID- 8403694 TI - Absence of hepatic uptake of Tc-99m phytate in a man with chronic toluene hepatotoxicity. AB - The case reported here is of a 28-year-old man diagnosed as having toluene hepatotoxicity. He had a 5-year history of exposure to toluene and was admitted to hospital complaining of two episodes of loss of consciousness. The high total bilirubin and the low prothrombin time suggested serious liver dysfunction. Based on histologic examination of a liver biopsy specimen, the diagnosis of toxic liver dysfunction caused by toluene poisoning was made. Hepatic images were not present in Tc-99m phytate scintigrams, but they were present in hepatobiliary scintigrams done with the Tc-99m N-pyridoxyl-5-methyltryptophan. The diagnosis of hepatic reticuloendothelial failure was made. PMID- 8403695 TI - Tc-99m sestamibi and other agents in the detection of metastatic medullary carcinoma of the thyroid. AB - The 10-year survival rate for medullary carcinoma of the thyroid (MCT) is 50%; thus, good tumor-seeking radiopharmaceuticals are needed to localize foci of recurrence and metastasis during follow-up. Two patients with metastatic MCT were studied with Tl-201, I-131 MIBG, Tc-99m (V)-DMSA, and Tc-99m MIBI. A SPECT study with the latter agent allowed the visualization and precise localization of a metastatic mediastinal lymph node. More studies need to be done to evaluate the role of Tc-99m MIBI in the detection of recurrence and metastases of MCT. PMID- 8403696 TI - Ablation of remaining functioning thyroid lobe with radioiodine after hemithyroidectomy for carcinoma. AB - Nineteen patients who had hemithyroidectomy as a surgical procedure for thyroid cancer were given radioiodine to ablate the residual hemithyroid. Two different protocols were used, with either a larger single dose or multiple smaller doses. Ablation was achieved in 28% with the single large dose approach and in 33% of patients given split doses of similar total amount. This response to the initial attempt at eradication of the residual normal hemithyroid is considerably lower than the accepted ablation rate with comparable doses administered to patients with small thyroid residues after total thyroidectomy. It is suggested that the effect of radioiodine treatment may relate inversely to the size of the thyroid remnant. Our findings also are in support of the surgical approach advocating total thyroidectomy for patients contemplating ablation of thyroid residues after surgery for thyroid carcinoma. PMID- 8403697 TI - Caroli's disease versus polycystic hepatic disease. Differential diagnosis with Tc-99m DISIDA scintigraphy. AB - Four patients with multiple hepatic cysts were studied: two were diagnosed with Caroli's disease (CD) and two with polycystic hepatic disease (PHD). In CD, hepatic scintigraphy with Tc-99m DISIDA showed areas of focally increased radiotracer accumulation that persisted more than 120 minutes, whereas in PHD, areas of focally decreased radiotracer accumulation were observed with normal liver washout and biliary excretion. When multiple hepatic cysts are shown by abdominal echography or CT scan, hepatic scintigraphy with Tc-99m DISIDA should be performed. This examination is safe and noninvasive, and permits differential diagnosis between CD and PHD. PMID- 8403698 TI - The complementary role of hepatobiliary scintigraphy and abdominal sonography in a patient with a hepatic cyst and suspected acute cholecystitis. AB - A 51-year-old woman was admitted with suspected acute cholecystitis. A large hepatic cyst was found incidentally by abdominal sonography, which displaced an "abnormal" gallbladder. Hepatobiliary scintigraphy visualized the gallbladder and excluded acute cholecystitis, but required the anatomic information from sonography to verify the abnormal location of the gallbladder. Additionally, scintigraphy showed the cyst not to communicate with the biliary tract. Pathologic findings revealed mild chronic cholecystitis and a simple mesothelial hepatic cyst. This case illustrates the complementary role of hepatobiliary scintigraphy and abdominal sonography in patients with distorted hepatobiliary anatomy and suspected acute cholecystitis. PMID- 8403699 TI - Limitations of quantitative radionuclide bone scanning in the evaluation of total knee replacement. AB - To establish the value of quantitative radionuclide bone scanning after total knee replacement, 21 patients were scheduled for examination. Eight patients were scanned more than 1 year after surgery (group 1), and 13 within the first postsurgical year (group 2). Ratios were calculated for three regions of interest. The ratio was defined as the counts per pixel for each region of interest, normalized by dividing them by the counts per pixel of the ipsilateral femoral shaft. There was no significant difference in uptake between control subjects and patients in group 2, indicating that the test was not reliable within this time period. No statistical difference was found between the uptake scores in groups 1 and 2, demonstrating that an uninterpretable period of 1 year is perhaps too short. Using a ratio of more than three, seen more than 1 year after surgery as pathologic, a sensitivity of 100%, a specificity of 33%, and an accuracy of 75% was found. Quantitative bone scanning is a very sensitive test for detecting complications, but it is nonspecific in diagnosing loosening of total knee arthroplasties. The value of quantitative scanning presumably lies in the follow-up of patients as an objective method for the evaluation of sequential scanning. Nevertheless, radionuclide data need to be interpreted in correlation with clinical and radiologic findings. PMID- 8403700 TI - SPECT imaging in a case of primary respiratory tract amyloidosis. AB - SPECT findings in a very rare case of primary amyloidosis localized in the laryngotracheobronchial area are reported. SPECT using Tc-99m PYP revealed widespread uptake in the larynx and the entire tracheobronchial tree up to the subsegmental divisions; the areas corresponded to diffuse thickening and calcification of the walls on CT. SPECT using Ga-67 citrate also showed marked uptake in the same area, consistent with the findings shown by SPECT using Tc-99m PYP. PMID- 8403701 TI - Segmental contour pattern in a case of pulmonary venoocclusive disease. AB - A V/Q mismatch (segmental contour pattern) is described in a patient suffering from dyspnea. Chest roentgenography revealed a mild interstitial pattern and enlarged pulmonary arteries. A lung biopsy demonstrated pulmonary venoocclusive disease. PMID- 8403702 TI - Discordance between F-18 fluorodeoxyglucose uptake and contrast enhancement in a brain abscess. AB - A 29-year-old human immunodeficiency virus-negative intravenous drug abuser with a right hemispheric staphylococcal brain abscess underwent PET imaging of 18F fluorodeoxyglucose (FDG) brain uptake. Diffuse right cortical hypometabolism was noted, as well as crossed cerebellar diaschisis. Unlike previous PET findings on FDG uptake in a brain abscess, however, the most intense uptake of FDG was found within the abscess and not in the contrast-enhancing walls. PMID- 8403703 TI - Role of In-111 labeled CYT-103 immunoscintigraphy in the evaluation of patients with recurrent colorectal carcinoma. AB - An immunoconjugate of the whole antibody, B72.3, has been approved by the FDA. It is labeled with indium-111, which allows imaging studies intended for the diagnosis and staging of colorectal and recurrent ovarian carcinoma. The new diagnostic imaging agent (CYT-103), Oncoscint CR/OV, has found particular use in evaluating patients with recurrent carcinoma. The three major indications for the study are: 1) occult disease--unexplained rise in serum tumor markers (e.g. CEA), with negative work-up including CT; 2) known local recurrence (e.g. liver or rectal area)--surgery is planned, but there is a need to rule out other areas of involvement; 3) clarify equivocal CT or MRI finding--to distinguish whether an abnormality is due to recurrent tumor or scar tissue, fibrosis, or unopacified bowel loop. Oncoscint has proven to be more sensitive than CT in the detection of disease in the pelvis and extrahepatic abdomen. CT remains the modality of choice for the detection of liver metastases. The combined sensitivity of the two modalities is 88% on a per-patient basis. PMID- 8403704 TI - Focal nodular hyperplasia of the liver. Diagnosis by dynamic and SPECT scintigraphy. PMID- 8403705 TI - Bone scintigraphy in oxalosis. PMID- 8403706 TI - Liver scan in Budd-Chiari syndrome. Correlation with CT and Doppler. PMID- 8403707 TI - Unexpected brain uptake in radioimmunoscintigraphy using F(ab)2 monoclonal antibodies. PMID- 8403708 TI - Bell-clapper appearance in scrotal hematoma. PMID- 8403709 TI - Gallbladder uptake simulating hydronephrosis on Tc-99m MAG3 scintigraphy. PMID- 8403710 TI - Regenerating nodules associated with hepatic metastasis. PMID- 8403711 TI - CPR-induced rib fractures. Characteristic scintigraphic appearance. PMID- 8403712 TI - Spontaneous arteriovenous fistula secondary to abdominal aortic aneurysm detected by radionuclide angiography. PMID- 8403713 TI - Tc-99m RBC scan findings of gangrenous small and large bowel. PMID- 8403714 TI - Investigation of measures to reduce interfering abdominal activity on rest myocardial images with Tc-99m sestamibi. AB - With the new myocardial agent Tc-99m sestamibi, relatively high abdominal uptake represents a major limiting factor. The effect of a standard feeding (commercial milkshake taken immediately after injection), and posture (standing versus sitting for 10 minutes postinjection) on the resting biodistribution of sestamibi was investigated in patients receiving 3 to 5 MBq/kg injections as part of rest stress tomography. Ancillary anterior 1-minute images of heart and abdomen were obtained at 15 minutes postinjection and 90 to 140 minutes postinjection in 32 patients, randomized to feeding and postural treatments. Feeding decreased the activity in the gallbladder at both 15 and 110 minutes, but had no effect on liver parenchyma; activity in a background rectangle immediately beneath the heart was decreased by feeding only on the 15-minute images. An effect of posture was not apparent. Further study of the acute effects of a milkshake in eight patients showed a prompt reduction of 26% in a subdiaphragmatic background rectangle, but a more gradual decline in gallbladder counts. Thus, feeding of lipid after injection is not an essential component of sestamibi imaging protocols; oral administration of fluid immediately before imaging may help reduce interfering gastric activity. PMID- 8403715 TI - Hepatobiliary excretion of MAG3. Simulation of a urinary leak. AB - A Tc-99m MAG3 renal scintigram was performed on a patient with severe renal insufficiency and bilateral ureteral stents. The Tc-99m MAG3 study displayed typical findings for renal insufficiency but also suggested the presence of lower tract obstruction. Moreover, the delayed posterior and posterior oblique views of the abdomen revealed an extrarenal collection of tracer suggestive of a urinary leak. With the help of additional views, this extrarenal activity was proven to be tracer in the bowel and biliary system. Biliary excretion of Tc-99m MAG3 can be prominent in patients with renal insufficiency and could lead to an erroneous diagnosis of urinary leak. Finally, scintigraphic diagnosis of lower tract obstruction may be difficult in patients with renal insufficiency, but some helpful signs may lead to the correct diagnosis. PMID- 8403716 TI - Hiatal hernia on whole-body radioiodine survey mimicking metastatic thyroid cancer. AB - Total body I-131 scanning done on a 67-year-old woman with thyroid cancer revealed abnormal tracer uptake within the mid-thorax, thought to represent metastasis in the mediastinum or thoracic spine. Single-photon emission computed tomography images of the thorax showed contiguity between the thoracic uptake and the normal, physiologic uptake of radioiodine in the stomach, suggesting physiologic accumulation within a hiatal hernia. To confirm the alimentary tract location of the radioiodine, a study using orally administered Tc-99m SC in water was performed. The images were similar to the iodine images and revealed a pattern consistent with hiatal hernia. A hiatal hernia was also observed as an incidental finding on chest radiographs and an MRI. This case illustrates the potential for an abnormal anatomic configuration to mimic metastatic thyroid cancer and shows how SPECT and oral Tc-99m SC images may be useful in making this distinction. PMID- 8403717 TI - Comparative SPECT study of stroke using Tc-99m ECD, I-123 IMP, and Tc-99m HMPAO. AB - Brain perfusion SPECT imaging using Tc-99m ethyl cysteinate dimer (ECD) was carried out in 12 patients with chronic cerebrovascular disease. The sensitivity of lesion detection and lesion-to-normal tissue contrast was compared with those of I-123 IMP and Tc-99m HMPAO. Eight patients underwent all three studies and the remaining four patients had Tc-99m ECD and I-123 IMP scans. The sensitivity of lesion detection and lesion contrast was analyzed quantitatively using an asymmetric index determined by the equation: 100 x [right - left[/(right + left). In the cerebral cortex, cerebellum, and white matter, I-123 IMP showed higher lesion sensitivity than Tc-99m ECD and Tc-99m HMPAO. In the striatum and thalamus, Tc-99m ECD and I-123 IMP showed higher lesion sensitivity than Tc-99m HMPAO. I-123 IMP showed the highest lesion contrast in cerebral cortex and cerebellum, whereas Tc-99m ECD showed the highest contrast in thalamus and striatum. In all regions, Tc-99m ECD showed higher lesion contrast than Tc-99m HMPAO. These results suggest regional variation in the sensitivity of lesion detection and lesion contrast of Tc-99m ECD compared to I-123 IMP, and the superiority of Tc-99m ECD to Tc-99m HMPAO. PMID- 8403718 TI - A discrepancy between bone scan and MRI concerning the involvement of adjacent bone in soft tissue sarcoma. AB - The case of a patient with malignant fibrous histiocytoma is described. Emphasis is placed on the preoperative implementation of bone scintigraphy and MRI, which yielded discrepant findings. The ultimate therapeutic approach is discussed in light of the discrepancy and the conclusive pathologic result. PMID- 8403719 TI - Combined transmission-emission imaging in lymphoscintigraphy. AB - A modification of standard lymphoscintigraphic imaging is described that incorporates the use of a two-headed camera with flood source to provide combined whole-body transmission-emission images of lymphatic drainage in a patient with malignant melanoma. The most common imaging technique for lymphoscintigraphy uses single-head gamma cameras with or without skin markers. The authors performed both techniques on their patient, and believe that the modification may provide a clearer image for lymphoscintigraphic interpretation. PMID- 8403720 TI - Unexpected pulmonary clumping of In-111 leukocytes attributable to indium oxine reagent. AB - Five cases of artifactual In-111 leukocyte pulmonary activity were noted at three local hospitals in a 4-day period. Based on the differences in the preparation of the final indium leukocyte product, the problem could be attributed to the indium oxine reagent in a specific lot. This artifact of multiple small foci of marked increased activity in the lungs (clumping pattern), attributable to the In-111 oxine reagent, has not been described previously. PMID- 8403721 TI - Ictal SPECT in a 16-day-old infant. AB - Neonatal seizures can be difficult to classify according to partial versus generalized onset on the basis of clinical appearance or electroencephalography (EEG). Single-photon emission computed tomography has proven to be useful in adults when adjunctive tests are needed to identify the nature of seizure onset. Although its use has been extended recently to children, the lower age limit at which this technique is useful remains to be established. A case is reported in which ictal Tc-99m HMPAO SPECT of a 16-day-old infant revealed an area of focal hypermetabolism in the right temporal lobe corresponding to an area of focal atrophy revealed by MRI. The EEG of this infant demonstrated multifocal interictal epileptiform abnormalities and an ictal pattern with a generalized onset. This case indicates that ictal SPECT is a useful tool in the evaluation of even the youngest patients with seizures. PMID- 8403722 TI - Preliminary Tl-201 SPECT for assessment of treatment efficacy in three patients with pancreatic cancer. AB - To evaluate treatment efficacy, Tl-201 SPECT was performed in three patients with pancreatic cancer. In all of the patients, the degree of Tl-201 uptake by tumor, evaluated from the tumor-liver uptake ratio of Tl-201 activity, correlated well with alterations in serum level of a tumor marker (CA 19-9, DUPAN-2) after treatment. One of the patients showed no tumor shrinkage on CT after treatment; however, Tl-201 SPECT demonstrated reduced uptake in the tumor coupled with a decreased level of CA 19-9. These results suggest that Tl-201 SPECT may become a new tool for assessing the efficacy of treatment in pancreatic cancer. PMID- 8403723 TI - The motility of interposition in patients with esophageal carcinoma after reconstructive esophageal surgery. AB - This study evaluated the motility of interposition in patients with esophageal carcinoma after different reconstructive operations of the esophagus using an esophageal radionuclide transit test. A modified standard method was used to calculate total mean transit time (MTT), residual fraction (RF), and retrograde index (RI) in the supine position at least 6 months after the reconstructive operations of the esophagus. Eleven patients with esophagogastrostomy (EG), 7 patients with esophagoiliocolostomy (EIC), and 25 normal volunteers with a similar distribution of ages were included in this study for comparison. The values of the scintigraphic parameters in patients with esophageal reconstruction were significantly worse than in normal volunteers (P < 0.05). In addition, the MTT and RF in patients with EG were higher than in patients with EIC (P < 0.05), but no significant difference in RI was demonstrated. In conclusion, the motility of interposition became worse after reconstructive operations, but reconstruction with the colon can preserve better motility than with the stomach. PMID- 8403724 TI - Oversized "bullseye" with central activity. A scintigraphic appearance of hemorrhagic testicular infarct following traumatic rupture of the testicle. PMID- 8403725 TI - A renal transplant seen on dual photon absorptiometry. PMID- 8403726 TI - Diffuse idiopathic skeletal hyperostosis can resemble metastases on bone scan. PMID- 8403727 TI - Chemotherapy-related resolution of radiogallium uptake in a lactating adenoma. PMID- 8403728 TI - Skeletal imaging in corrected pectus excavatum. PMID- 8403729 TI - Tc-99m renal scan demonstrating growth in a pediatric renal transplant. PMID- 8403730 TI - Bone agent and radiogallium deposition around infiltrated calcium gluconate. PMID- 8403731 TI - Interruption of aortic flow between the thoracic and abdominal aorta with development of collateral circulation secondary to bronchogenic carcinoma. PMID- 8403732 TI - Metastatic cardiac liposarcoma diagnosed by thallium-201 myocardial single photon emission computed tomography. PMID- 8403733 TI - The "batwing" thyroid gland. A case of thyroid carcinoma. PMID- 8403734 TI - Clinical pharmacokinetics of drugs in obesity. An update. AB - Obesity is common enough to constitute a serious medical and public health problem. Drug prescription for obese patients is difficult since dosages based on pharmacokinetic data obtained in normal-weight individuals could induce errors. In obese patients, physiopathological modifications are likely to affect drug tissue distribution and elimination. Body constitution is characterised by a higher percentage of fat and a lower percentage of lean tissue and water. Although the cardiac output and total blood volume are increased, the blood flow per gram of fat is less than in nonobese individuals. Histological hepatic alterations are commonly reported in morbidly obese individuals. A higher glomerular filtration rate is also observed. Most of the pharmacokinetic information concerning obesity deals with distribution. Published data concerning molecules with moderate and weak lipophilicity are homogeneous. In obese compared with normal weight individuals, the total volume of distribution (Vd) is moderately increased (aminoglycosides, caffeine) or similar (H2-blockers, neuromuscular blockers), but the Vd corrected by kilogram of actual bodyweight is significantly smaller. These drugs distribute to a limited extent in excess bodyweight. For highly lipophilic drugs, despite this common characteristic, discrepancies in distribution in obesity exist between drugs belonging to different pharmacological classes. Some drugs show a clear augmentation of Vd and elimination half-life (benzodiazepines, carbamazepine, trazodone, verapamil, sufentanil), indicating a marked distribution into adipose tissue. For others, Vd and Vd/kg are decreased (cyclosporin, propranolol), suggesting that factors other than lipid solubility intervene in tissue distribution. As a general trend, the total clearance (CL) of drugs metabolised by oxidation, conjugation or reduction, and also of drugs with flow-dependent hepatic clearance, is not diminished in obesity. Usually CL is identical in obese and nonobese individuals, sometimes it is increased in obesity (enflurane, halothane, prednisolone, some benzodiazepines). With some drugs a significant reduction in CL is observed in obese individuals (methylprednisolone, propranolol). Renal clearance of aminoglycosides and cimetidine increases in obese individuals. Practical guidelines for dosage adjustment are proposed. For drugs with distribution restricted to lean tissues, the loading dose should be based on the ideal bodyweight of patients. For drugs markedly distributed into fat tissue the loading dose is based on total bodyweight. Adjustment of the maintenance dose depends on possible changes in CL. In some cases (atracurium, prednisolone) dosage adjustment does not follow these recommendations, owing to pharmacodynamic data. PMID- 8403735 TI - Protein binding and stereoselectivity of nonsteroidal anti-inflammatory drugs. AB - Stereoselective binding of nonsteroidal anti-inflammatory drugs (NSAIDs) can be studied using various techniques. Thus the results obtained by different investigators may be poorly consistent and even contradictory. NSAIDs are bound stereoselectively to serum albumin to different degrees depending on the drug investigated (ibuprofen, indoprofen, carprofen, etodolac, ketoprofen and flurbiprofen). For other drugs, both enantiomers are bound to a similar extent (pirprofen, fenoprofen). This stereoselectivity could vary with experimental conditions, in particular with protein concentration (ketoprofen, etodolac), leading to individual differences. Finally, the stereoselectivity of protein binding and of pharmacokinetics can be compared: differences in binding between enantiomers can explain their differences in pharmacokinetics, once metabolic properties such as inversion have been taken into account. PMID- 8403736 TI - Pharmacokinetic optimisation of inhaled steroid therapy in asthma. AB - The recognition that asthma has a large inflammatory component has led to the use of steroids in its treatment. The adverse systemic effects of the drugs have promoted the development of inhaled steroids with a high topical to systemic potency ratio. The 2 most widely used agents are beclomethasone dipropionate and budesonide. Budesonide has a longer plasma elimination half-life than beclomethasone dipropionate but a higher topical to systemic potency ratio. These agents have been shown to be equipotent with respect to their anti-asthma effects but budesonide may have a slightly more favourable adverse effect profile. At low dosages (up to 400 micrograms/day) these drugs are well tolerated. At higher dosages (> 1000 micrograms/day) adverse effects on bone metabolism and adrenal function have been noted. Other agents such a triamcinolone or flunisolide have no obvious advantages. We recommend that inhaled steroids should be prescribed at the lowest dose required to control symptoms, with the dose being increased or decreased in a stepwise manner in parallel with asthma severity. PMID- 8403737 TI - The relationship between phenazone (antipyrine) metabolite formation and theophylline metabolism in healthy and frail elderly women. AB - The influence of aging on the metabolism of phenazone (antipyrine), and the relationship between the formation of 3 phenazone metabolites and the metabolic clearance of theophylline in healthy and frail elderly women, were examined. Whereas the elimination half-life did not change, clearance of phenazone decreased by about 50% with age in healthy women receiving phenazone without theophylline. However, the summation of the urinary recovery of phenazone and the measured metabolites, expressed as percentage of the phenazone dose, was lower in the healthy elderly (37 +/- 9% vs 74 +/- 15%). In both healthy and frail females the clearance of formation of 4-hydroxy-phenazone and the metabolic clearance of theophylline correlated strongly (r = 0.93 and 0.90, respectively). In non healthy elderly females, strong correlations were also observed between the other metabolic pathways of phenazone and the metabolic clearance of theophylline. Coadministration of theophylline in the elderly increased the percentage of the phenazone dose excreted as the measured metabolites. A considerably higher interindividual variability in the disposition of phenazone and theophylline was observed in the frail elderly women. This high degree of variability in drug metabolism may be one of the explanations for the problems often occurring after drug prescription in the elderly. PMID- 8403738 TI - Pharmacokinetics of epoetin (recombinant human erythropoietin) after long term therapy in patients undergoing haemodialysis and haemofiltration. AB - After long term therapy with epoetin (recombinant human erythropoietin) 17 patients with end-stage renal disease (ESRD) were studied in 3 groups to assess pharmacokinetics during the intertreatment interval and during haemofiltration and dialysis treatment. Epoetin was measured by radioimmunoassay. After an intravenous bolus of epoetin 150 U/kg bodyweight, the half-life was 7.7h, steady state volume of distribution was 0.066 L/kg and total plasma clearance was 5.4 ml/min. The mean steady-state serum concentration during multiple-dose administration was 656 U/L. The drug was not eliminated by haemofiltration or dialysis. Long term treatment of ESRD patients with epoetin does not significantly alter the pharmacokinetic profile of the drug. Epoetin dosage adjustment or substitution after haemofiltration and dialysis is not necessary. PMID- 8403739 TI - Disposition of ibuprofen in patients with liver cirrhosis. Stereochemical considerations. AB - Following a single oral dose of racemic ibuprofen 600mg the stereoselective disposition of its enantiomers was studied in 8 patients with moderate to severe cirrhosis. Compared with the elimination half-life (t1/2) of (-)-R- and (+)-S ibuprofen in 8 healthy age-matched controls (1.7 +/- 0.3h and 1.8 +/- 0.5h, respectively), t1/2 was prolonged significantly (p < 0.045 and < 0.001, respectively) in patients with cirrhosis (t1/2 = 3.1 +/- 1.7h and 3.4 +/- 1.0h, respectively). Whereas the low amounts excreted unchanged into urine differed slightly in both groups studied, much less (p < 0.01) conjugated ibuprofen was recovered either as the R-enantiomer (0.9 +/- 0.4% vs 4.1 +/- 2.8% of the dose) or the S-enantiomer (6.4 +/- 2.5% vs 26.5 +/- 12.9% of the dose) in patients with cirrhosis. Metabolic inversion of the inactive (-)-R-ibuprofen to the active (+) S-ibuprofen may be impaired in hepatic dysfunction since the normal ratio of areas under the curve (AUC) for R- and S-enantiomers (0.79 +/- 0.18) was significantly (p < 0.02) higher in patients with cirrhosis (1.10 +/- 0.28). In a second study, a single oral dose of 400mg (+)-S-ibuprofen was administered to 8 healthy volunteers and 8 patients with cirrhosis. Elimination of this enantiomer was slightly impaired as could be seen from the prolonged t1/2 (1.6 +/- 0.1h vs 2.6 +/- 0.5h; p < 0.001) and the increase in AUC (101 +/- 35 vs 144 +/- 41 mg/L.h; p = 0.041). In conclusion, in patients with liver disease, hepatic elimination of ibuprofen is impaired. This should be taken into consideration especially if the racemic drug is used. Direct administration of the active (+)-S enantiomer seems to offer less vulnerable treatment. PMID- 8403740 TI - Therapeutic drug monitoring in hair samples. Principles and practice. PMID- 8403741 TI - Cefepime clinical pharmacokinetics. AB - Cefepime is a new parenteral cephalosporin with antimicrobial activity similar to third-generation cephalosporins. It acts against the Enterobacteriaceae family, and Pseudomonas aeruginosa, but maintains Gram-positive activity similar to that of first- or second-generation cephalosporins. Cefepime has in vitro activity against many bacterial isolates resistant to ceftazidime and cefotaxime, is stable against chromosomally mediated beta-lactamases, demonstrates lower affinity for these enzymes and shows a high resistance to enzymatic hydrolysis. Clinical uses thus far include treatment of lower respiratory tract, intra abdominal and urinary tract infections, skin and soft tissue infections and for prophylaxis in biliary tract and prostate surgery. Pharmacokinetic studies indicate that cefepime exhibits linear pharmacokinetic behaviour. Pharmacokinetic variables are not significantly different between single- and multiple-dose administration, indicating a lack of drug accumulation in patients with normal renal function. Cefepime is not highly bound to plasma proteins, with binding values of approximately 16 to 19%. The drug is widely distributed in various biological tissues and fluids. The primary route of elimination is from the kidneys, with over 80% of the drug recovered in the urine as unchanged drug in patients with normal renal function. Total drug clearance and renal clearance are similar to creatinine clearance, and glomerular filtration is thought to be the primary mechanism of renal excretion. The elimination half-life is approximately 2 to 2.5 h in patients. Cefepime is removed by haemodialysis (over 3h) and peritoneal dialysis (over 72h) to an appreciable extent, with 40 to 68% and 26% of the drug removed, respectively. Overall, cefepime is well tolerated by patients and no significant drug interactions have been reported to date. PMID- 8403742 TI - Atypical extrapulmonary presentations of severe respiratory syncytial virus infection requiring intensive care. AB - The patterns and nature of a four-month epidemic of severe respiratory syncytial virus (RSV)-associated disease were analyzed using presenting, demographic, clinical, and therapeutic data. Of 218 infants with RSV infection admitted to Rainbow Babies and Childrens Hospital, 49 (22.4%), most born prematurely, entered the pediatric intensive care unit (PICU). Fluorescent antibody and/or enzyme linked immunosorbent assay documented RSV infection. PICU patients underwent airway stabilization; 53.5% were intubated and evaluated for sepsis. Patients with positive bacterial cultures received antibiotics; 18% were given ribavirin. Patterns of infection included hypothermia, septic shock appearance, apnea, pneumonia, and wheezing due to bronchiolitis. The average age difference between patients with hypothermia (23.3 days) and those with pneumonia (11.2 months) was statistically significant. There were no significant differences in average age, gestational age at birth, number intubated, worst pH and PCO2, duration of intensive care, or treatment modalities between infants with bronchopulmonary dysplasia who received ribavirin and those who did not. PMID- 8403743 TI - Respiratory syncytial virus infection: the expanded clinical spectrum. PMID- 8403744 TI - Early hospital discharge and the timing of newborn metabolic screening. AB - Cost containment measures have reduced dramatically the length of stay for normal newborns, in some cases jeopardizing the ability to obtain appropriate newborn screens. In our hospital, we found that an unacceptable number of patients had mistakenly been screened before 24 hours of age. As pressures to shorten hospitalization increase, health-care providers must examine the impact of such changes on their ability to obtain adequate newborn screens. Potential solutions include continued vigilance in gathering specimens after 24 hours of age, interpretation of time-sensitive tests in an age-adjusted manner, and repeating newborn screens after 24 hours of age. PMID- 8403745 TI - False-negative urine latex particle agglutination testing in neonates with group B streptococcal bacteremia. A function of improper test implementation? AB - Although blood cultures remain the most specific indicator of Group B streptococcus (GBS) sepsis, a potentially life-threatening infection in neonates, test results may not be available for 24 to 48 hours. Detection of GBS antigen in the urine by latex particle agglutination (LPA) may speed diagnosis. This study analyzed the sensitivity of the GBS urine LPA assay under clinical conditions. The urine of neonates with early-onset GBS bacteremia was analyzed for GBS antigen over a three-year period at six military medical centers. Overall, 53.5% (38/71) of infants with positive blood cultures had a positive urine LPA test. Only one medical center routinely followed manufacturer's recommendations to concentrate urine specimens before testing. These data suggest that the sensitivity for the urine LPA assay, when performed on unconcentrated urine, is lower than previously reported. Clinicians should insist that the laboratory maximize sensitivity by concentrating urine prior to GBS LPA testing. PMID- 8403746 TI - Reliability and validity of an infant gastroesophageal reflux questionnaire. AB - To improve history-taking of infants with suspected gastroesophageal reflux, we developed an Infant Gastroesophageal Reflux Questionnaire consisting of 161 items covering demographics, symptoms (regurgitation, weight deficit, respiratory difficulties, fussiness, apnea, and pain or bleeding of esophagitis), and possible causes (feeding volume and frequency, allergy, infection, colic, central nervous system abnormalities, positioning, and smoke exposure). The questionnaire was completed by primary caretakers of 69 infants aged 1 to 58 weeks suspected of having reflux. Median time to complete the questionnaire was 20 minutes. The median internal consistency of 29 pairs of redundant questions was 0.94. Median test-retest consistency of 110 items for nine respondents was 0.88. Median interobserver consistency, evaluated for 129 items in 35 questionnaires also filled out by secondary caretakers, was 0.85. The median accuracy of four externally validated items was 1.00. This questionnaire can aid pediatricians in making decisions regarding diagnoses and treatment in this common but complex disorder. PMID- 8403747 TI - Estimating the predictive value of a diagnostic test. How to prevent misleading or confusing results. AB - The diagnostic process consists of a series of steps in which the estimated probability of particular disease is increased or decreased until either treatment can be instituted or the diagnosis excluded. The history and physical exam play an important role. If a therapeutic decision cannot confidently be made on clinical grounds alone, a diagnostic test may confirm or exclude a diagnosis or clearly indicate the need for further testing. An inappropriately chosen test or misinterpreted test result, however, may mislead the clinician and possibly harm the patient or simply be wasteful. Estimating the predictive values of a diagnostic test will help to avoid these pitfalls. This article shows the clinician a simplified way to do this. PMID- 8403748 TI - Costochondritis in adolescents. A follow-up study. PMID- 8403749 TI - Contamination errors when sampling blood from an arterial line. PMID- 8403750 TI - Safety of ciprofloxacin in children with cystic fibrosis. PMID- 8403751 TI - Antibiotic dosages in the treatment of group A streptococcal infections. PMID- 8403752 TI - Perioperative management of pheochromocytoma in children. PMID- 8403753 TI - Methylphenidate protocol: feasibility in a pediatric practice. PMID- 8403754 TI - Should all children receive two measles vaccinations? PMID- 8403755 TI - Simultaneous Kawasaki disease in group A streptococcal pharyngitis. PMID- 8403756 TI - Evaluation and management of stridor in the newborn. PMID- 8403757 TI - Otitis media in practice. PMID- 8403758 TI - Staging of lymphoma in childhood. PMID- 8403759 TI - The future of interventional radiology--does it have one? PMID- 8403760 TI - The role of hepatic arterial embolization in the management of ruptured hepatocellular carcinoma. AB - Fifteen patients with ruptured hepatocellular carcinomas and intraperitoneal bleeding were considered for hepatic arterial embolization. Embolization was attempted in nine patients (60%) with patent or partially patent portal veins. Successful occlusion of the feeding tumour vessels was achieved in eight patients (53%) with six patients (40%) surviving more than 3 months compared to three survivors in the non-embolized group, all of whom had total occluded portal veins. Prognosis depends on the underlying liver function with total serum bilirubin levels of survivors significantly lower than those patients who died (P < 0.01). Embolization was tolerated well in those patients with partial portal vein occlusion. Selective tumour embolization should be the initial treatment of choice in these severely ill patients who have reasonable hepatic function. PMID- 8403761 TI - Local treatment of colorectal liver metastases: a comparison of interstitial laser photocoagulation (ILP) and percutaneous alcohol injection (PAI). AB - The purpose of this study was to evaluate the relative merits of two physical methods of locally destroying colorectal liver metastases-interstitial laser photocoagulation (ILP) which causes thermal necrosis, and percutaneous alcohol injection (PAI) which causes cellular dehydration and coagulative necrosis. Seventy-six liver metastases in 22 patients were treated by ILP or PAI. Both techniques were performed using local anaesthesia and intravenous sedation/analgesia. Ultrasound was used to localize the tumours and guide the needles percutaneously. ILP: Up to eight 19 G needles were inserted into the tumour, and down each needle was passed a thin optical fibre; the tumour was heated using low power laser light (2 W) for 500 s from a Nd:YAG or diode laser. PAI: 0.5-1 ml of sterile 95% absolute alcohol was injected into multiple sites of the tumour using a single 19-22 G needle. Dynamic CT scan was used to evaluate the extent of treatment-induced necrosis. Ultrasound showed echogenic changes around the needles/fibre-tips during ILP and PAI; this gave a reasonable guide to the extent of thermal damage for smaller tumours during ILP, but not during PAI. ILP: 54 tumours were treated (median size 2.7 cm). Laser-induced necrosis was clearly seen 24 h after treatment as a well-defined area of non-enhancement on the dynamic CT scan; greater than 50% necrosis of tumour volume was achieved in 87% of tumours (complete necrosis was found in 52% of tumours). PAI: 22 tumours were treated (median size 1.5 cm). Dynamic CT showed patchy areas of non enhancement in five tumours, decreased density in seven tumours, and no change in 10 tumours; complete tumour necrosis was never achieved. There were no major complications after ILP or PAI, but pain during treatment was more common and more severe with PAI. ILP is a simple, safe and effective treatment for colorectal liver metastases; PAI is relatively ineffective for these tumours (although it has been shown to be much more effective for small hepatocellular carcinomas). PMID- 8403762 TI - Radiological aspects of solvent dissolution of gall-stones. AB - Twenty-five patients were referred for solvent dissolution of gall-stones using methyl tertiary butyl ether (MTBE). All patients were assessed beforehand by ultrasound and oral cholecystography. Computed tomography was performed in 23 patients to assess stone calcification and gall-bladder anatomy. Successful stone dissolution was achieved in 15 patients and partial dissolution in six patients. Failure to catheterize the gall-bladder occurred in four patients. There were four biliary leaks, two requiring cholecystectomy. The CT appearances of the gall bladder may have predicted problems with catheterization in three of the four cases, and the results suggest that cover of the gall-bladder by the liver of greater than 50% is a favourable predictor of successful catheterization. A thick walled gall-bladder and excess fat between the liver and the lateral abdominal wall are adverse predictors of successful gall-bladder catheterization. Both tubography and ultrasound should be used to assess progress of dissolution. Newer, more effective solvents together with increased expertise in interventional techniques may encourage the more widespread use of solvent dissolution of gall-stones as an alternative to surgery in a selected population. PMID- 8403763 TI - Precontrast and postcontrast (Gd-DTPA) magnetic resonance imaging of hand joints in patients with rheumatoid arthritis. AB - In an attempt to demonstrate whether clinically selected joints of the hand in active rheumatoid disease had consistent MRI findings, 45 patients were examined, in whom one joint in each was selected by both the referring clinician and patient as being active and symptomatic. Such joints, in order to be included in the study, were required to conform to ARA criteria of activity and usually mild to moderate X-ray changes. The joints were imaged using spin-echo sequences with T1W and T2W precontrast images, followed by T1W images after intravenous administration of Gd-DTPA. Different patterns of joint abnormalities were found. In 27 joints MRI findings suggested highly active synovitis and/or destructive pannus. In four, crescentic enhancement was thought to be compatible with simple synovitis, but in 23 rounded masses of synovial proliferation were characterized by marked, diffuse contrast enhancement on T1W postcontrast images, which corresponded well with high signal intensity on T2W images. Synovial proliferation in a further 12 joints was shown by only moderate stippled contrast enhancement and nonhomogeneous intermediate to high signal intensity on T2W images. These findings were thought to represent less active synovitis and pannus. MRI did not demonstrate inflammatory activity in six joints. In two of these pannus was of low signal intensity on T2W images, without contrast enhancement after Gd-DTPA infection presumed fibrotic and inert, and four were normal on all pulse sequences. These results suggest that clinical features of synovitis, even in carefully selected joints clinically, do not produce a homogeneous group when examined by MRI imaging. Indeed, a spectrum exists from presumed marked, active synovitis to total normality. If MRI is to be used as a clinical and research tool in the assessment of rheumatoid disease, and its therapeutic manipulation, these results are of some importance, since the variable findings indicate an appreciable heterogeneity of appearances in joints thought clinically to be of relatively uniform severity. PMID- 8403764 TI - Bolus chasing: a new technique in peripheral arteriography. AB - Recent advances in digital angiographic equipment design have enabled a new technique of peripheral arteriography involving digital bolus chasing to be developed. This technique permits real-time visualization of the contrast bolus so that it can be followed peripherally with digital images being acquired at a suitable frame rate. We give the first description of this technique using the Philips Integris C2000 and compare it to conventional stepped arteriography. Sixty-one patients undergoing peripheral arteriography using the conventional stepped technique were compared with 55 patients using bolus chasing. The parameters assessed were the procedure time, the number of runs and screening time per investigation, the consumables used, the patient and radiologist radiation dose and the image quality. A reduction in the procedure time and the patient and radiologist radiation dose, and improvement in image quality are demonstrated with the bolus chasing technique. Bolus chasing represents an important advance in peripheral arteriography. PMID- 8403765 TI - Superficial siderosis of the central nervous system--diagnosis by magnetic resonance imaging. AB - Superficial siderosis of the central nervous system is a rare disorder characterized by the deposition of haemosiderin in the leptomeninges, cranial nerves and spinal cord. The clinical features include sensorineural deafness, ataxia and pyramidal tract signs. We describe two cases demonstrating the characteristic findings on magnetic resonance imaging of widespread peripheral low signal on T2-weighted images. PMID- 8403766 TI - Effective dose during screening monitored intussusception reduction. AB - A retrospective survey of 58 intussusception reductions in 55 patients is presented. During the period studied, the department changed from the use of barium to air as a reduction medium. The reduction rates were 86% with air (37 procedures) compared to 67% with barium (21 procedures). The complication rates were similar. The use of a dose-area product meter enables effective dose to the patient to be assessed. The median effective dose to the patient is 55 microSv, and the 75 percentile dose is 75 microSv. A table to estimate effective dose to the patient during the procedure is presented. This study confirms previously quoted advantages of pneumatic reduction over hydrostatic reduction of intussusception but finds no significant difference in dose between air and barium reduction. We believe that it should be possible to submit the patient to a dose of 75 microSv or less during intussusception reduction under screening control. PMID- 8403767 TI - Partial obliteration or blurring of the descending aortic contours: a pitfall on plain chest radiographs. AB - We report the cause and clinical significance of partial obliteration or blurring of the descending aortic contour on the frontal chest radiographs of patients without pathology in the vicinity of the descending aorta. Among 112 cases, 26 cases (23.2%) showed obliteration or blurring of the contour (positive group). On the corresponding computed tomograph of the positive group, two dominant causes were shown. One was contact between the descending aorta and the left hilar or lower lobe vessels. The second was the adjacent pleura being obliquely orientated to the anteroposterior (A-P) axis. In the latter group, on the lateral chest radiographs, a flattened thorax with a smaller A-P diameter was seen. Identification of left hilar or lower lobe vessels on a frontal chest radiograph or a diminished A-P diameter of the thorax on a lateral radiograph is useful in differentiating these circumstances from true thoracic pathology. PMID- 8403768 TI - Case report: Mirizzi syndrome--treatment with metallic endoprosthesis. PMID- 8403769 TI - Pictorial review: the sonographic diagnosis of pathology in the Achilles tendon. AB - This pictorial essay details Achilles tendon pathology using real-time sonography. Characteristic diagnostic images for tenosynovitis, acute and chronic tendinitis, peritendinitis, nodular tendinitis and complete and partial tendon ruptures are described. The anatomy of the Achilles tendon in relation to these diagnoses is discussed. Ultrasound is recommended as a useful first-line approach in imaging the Achilles tendon. PMID- 8403770 TI - Case report: azygos/accessory hemiazygos continuation of the inferior vena cava mimicking dissection of the aorta. AB - A case of azygos/accessory hemiazygos continuation of the inferior vena cava is presented. This mimicked the chest radiograph appearances of dissection of the aorta, a diagnosis suspected clinically. Contrast-enhanced CT enabled the correct diagnosis to be made. PMID- 8403771 TI - Case report: biliary peritonitis following Wallstent insertion. AB - A case is described of biliary peritonitis following insertion of a Wallstent endoprosthesis. Features of the case that may have contributed to this complication are described and methods of preventing such an outcome discussed. PMID- 8403772 TI - Case report: an adjunct to laparoscopic splenectomy--a new role for interventional radiology. PMID- 8403773 TI - Case report: superior vena caval obstruction treated by thrombolysis, mechanical thrombectomy and metallic stents. AB - We describe a case of superior vena caval obstruction (SVCO) due to bronchial carcinoma which was complicated by extensive central venous thrombosis. Partial clot lysis was achieved with thrombolytic agents, but therapy had to be discontinued due to bleeding from the gastrointestinal tract and puncture sites. Clot dissolution was completed using the Amplatz Thrombectomy Device, and the subsequent placement of a Gianturco Z-stent resulted in prolonged symptomatic relief. PMID- 8403774 TI - Extending the range of per-rectal needle drainage of pelvic sepsis. PMID- 8403775 TI - Transaxillary arteriograms. PMID- 8403776 TI - Ablation of mediastinal parathyroid adenomas. PMID- 8403777 TI - Mammographic accuracy and patient age. PMID- 8403778 TI - Problems with stopping rules in the trials of risky therapies: the case of warfarin to prevent stroke in atrial fibrillation. PMID- 8403779 TI - A novel recombinant adenovirus vector expressing a flavivirus non-structural protein protects against lethal flavivirus challenge. PMID- 8403780 TI - Renal functional response to ischaemic renal failure in chronic hypoxic rats. AB - 1. Chronic hypoxic rats are always polycythaemic. It is possible that an increase in packed cell volume may enhance erythrocyte trapping with a consequent increase in renal damage after renal ischaemia. These experiments were designed to assess renal functional changes after renal arterial occlusion in chronic hypoxic rats. 2. Chronic hypoxic rats were prepared by exposure (15h/day) to an altitude chamber (5486m) for 4 weeks. 3. After 45 min of left renal arterial occlusion, there were significant decreases in the excretion of potassium, p-aminohippurate and inulin and in the p-aminohippurate extraction ratio in 12 sea level ischaemic insulted kidneys. In 12 chronic hypoxic rats, the same parameters were changed after left renal ischaemia but only the p-aminohippurate ratio was significantly altered. 4. Administrations of 1 or 5 mg/kg phosphoramidon did not cause any significant improvement in the measured renal parameters in both kidneys and in both groups of rats after ischaemia. 5. In the second experiment, the rats were challenged by rapid infusion of 10 ml of saline intravenously, and urine was collected for 90 min from each ureter. Four hours after left renal arterial occlusion, the insulted kidney showed increased water and sodium excretion in both sea level and chronic hypoxic rats. However, 24 h after left renal ischaemia, the responses of sea level and chronic hypoxic rats were different. Urinary excretion was significantly reduced in sea level rats, but was almost normal in chronic hypoxic rats. 6. This report suggests that some beneficial factors after chronic hypoxia might play important roles in reducing the damage after renal ischaemia. PMID- 8403781 TI - A novel computer-driven, servo-controlled fluid replacement technique and its application to renal function studies in conscious rats. AB - 1. A new rat model has been developed allowing body fluid status to be accurately controlled and maintained throughout experimentation by computer-driven, servo controlled replacement of spontaneous urinary fluid losses. 2. Experiments in vitro were performed to test the accuracy of the servo system, and experiments in vivo were carried out to re-assess basic renal function in servo-controlled vasopressin-replete Long Evans and vasopressin-deficient Brattleboro rats. The model was further evaluated in water-diuretic Wistar rats with or without administration of a vasopressin V2-receptor agonist, 1-desamino-8-D-arginine vasopressin. 3. The data gained from the present study indicate the suitability of the servo-controlled replacement system for conscious renal function studies in three different rat strains. Haemodynamic and renal function variables measured were demonstrated to be stable throughout a 5 h experimental procedure and reproducible between repeated experimental occasions over a 14 day post operative period. 4. Using the servo-control technique, the expected action of 1 desamino-8-D-arginine vasopressin on renal water handling was demonstrated, but the natriuretic effect reported by some workers was not evident. 5. Since the servo-controlled fluid replacement technique maintains many of the inherent differences between vasopressin-replete Long Evans and vasopressin-deficient Brattleboro rats and eliminates the changes in body fluid volume during transition from a diuretic to an antidiuretic state, the model confers an advantage over previously employed constant infusion protocols. PMID- 8403782 TI - Reduced renal papillary plasma flow in non-ascitic cirrhotic rats. AB - 1. The purpose of the present investigation was to determine whether an abnormality of the renal papillary circulation is present in a well-established model of cirrhosis without ascites (carbon tetrachloride/phenobarbital). 2. Compared with the control animals, cirrhotic rats showed a reduced diuretic (61.0 +/- 5.1 versus 18.0 +/- 2.5%) and natriuretic (67.8 +/- 8.3 versus 29.6 +/- 3.6%) response to a volume expansion (3% body weight infusion of 0.9% NaCl). The volume expansion-induced increase in renal interstitial hydrostatic pressure was also blunted in the cirrhotic rats (control 9.3 +/- 0.9 versus cirrhotic 6.1 +/- 1.0 mmHg) and there were no differences in mean arterial blood pressure, renal blood flow or glomerular filtration rate between control and cirrhotic animals. 3. Papillary plasma flow was determined by the 125I-albumin accumulation technique and expressed as ml min-1 100 g-1. In the basal state, papillary plasma flow was significantly lower in cirrhotic rats (59.1 +/- 4.4, n = 9) than in the control animals (81.8 +/- 6.9, n = 9). An isotonic saline expansion similar to the one described above significantly increased papillary plasma flow in control rats (108.4 +/- 9.1, n = 7) but did not change it in cirrhotic rats (60.2 +/- 4.9, n = 6). 4. Our results indicate the existence of a selective alteration in the renal papillary circulation in cirrhotic rats, both in the basal state and after a well established vasodilatory stimulus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403783 TI - Circulating angiotensin II and renal sodium handling in man: a dose-response study. AB - 1. Animal studies have shown that angiotensin II has a biphasic effect on urinary sodium excretion. To examine whether this is also true in man, we studied seven salt-replete male subjects in a single-blind placebo-controlled manner. 2. While undergoing maximum diuresis, subjects were infused with 0, 1, 2, 5 or 10 ng of angiotensin II min-1 kg-1 over 80 min. Subjects were studied while seated, and stood every 20 min for urine collection. 3. Angiotensin II produced a dose dependent antidiuretic effect. The urine flow rate, in ml/min expressed as the change from baseline with increasing dose of angiotensin, was: +3.4 +/- 1.77, 1.26 +/- 0.49 (P < 0.05), -2.75 +/- 1.23 (P < 0.05), -4.21 +/- 0.82 (P < 0.05) and -6.51 +/- 1.07 (P < 0.01). 4. In contrast, the effect of angiotensin II on sodium excretion showed a flat dose-response curve beyond 5 ng min-1 kg-1. The urinary sodium excretion, in mumol/min expressed as the change from baseline with increasing dose of angiotensin, was: 9.5 +/- 21.2, -18.9 +/- 29.6, -37.0 +/- 11.6 (P < 0.05), -67.7 +/- 19.6 (P < 0.01) and -63.8 +/- 14.3 (P < 0.01). 5. The fractional distal reabsorption of sodium, determined by using the lithium clearance technique, showed a rise with all doses of angiotensin II used and reached statistical significance with the top two doses. 6. Unlike antidiuresis, antinatriuresis after graded doses of angiotensin II in human subjects showed a flat dose-response curve beyond 5 ng min-1 kg-1. Pressor doses of angiotensin II also have a significant effect on the distal tubule in promoting sodium reabsorption. PMID- 8403784 TI - Haemodynamic and hormonal responses to losartan (DuP753/MK954) infusion during cardiac catheterization in conscious salt-deplete dogs. AB - 1. Haemodynamic and hormonal responses to infused angiotensin II were studied in conscious salt-deplete dogs during infusion of D-glucose or losartan (DuP753/MK954). 2. Mean arterial pressure (118 +/- 13 mmHg) fell rapidly after losartan (60 min 106 +/- 18 mmHg) with a rise in heart rate (107 +/- 16 beats/min) from baseline (98 +/- 17 beats/min). Pressor responses to angiotensin II during D-glucose infusion (6 ng min-1 kg-1, 99 +/- 10 mmHg; 18 ng min-1 kg-1, 140 +/- 15 mmHg; 54 ng min-1 kg-1, 157 +/- 12 mmHg; 162 ng min-1 kg-1, 178 +/- 14 mmHg) showed a parallel shift during losartan infusion with very similar pressures in response to higher rates of angiotensin II infusion (54 ng min-1 kg 1, 108 +/- 17 mmHg; 162 ng min-1 kg-1, 138 +/- 14 mmHg; 486 ng min-1 kg-1, 155 +/ 14 mmHg; 1458 ng min-1 kg-1, 177 +/- 12 mmHg). Losartan caused a fall in baseline systemic vascular resistance. Despite the similar mean arterial pressure, the rise in systemic vascular resistance after angiotensin II during D glucose infusion (162 ng min-1 kg-1, 8065 +/- 1967 dyn s cm-5) was reduced during losartan infusion (1458 ng min-1 kg-1, 6645 +/- 1720 dyn s cm-5. Losartan caused a small rise in cardiac output related to a rise in heart rate and increased stroke volume. Pressure infusions of angiotensin II caused a fall in cardiac output during D-glucose infusion, which was blocked during losartan infusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403785 TI - Atrial natriuretic peptide in the pericardial fluid of patients with heart disease. AB - 1. The pericardial fluid of 20 open heart surgery patients with acquired heart disease was analysed for atrial natriuretic peptide by radioimmunoassay. 2. The concentration of atrial natriuretic peptide in the pericardial fluid was significantly higher than in the corresponding plasma (316.8 +/- 50.0 versus 121.7 +/- 29.1 pg/ml; P < 0.01) and was higher in patients with congestive heart failure than in those without heart failure (469.3 +/- 78.6 versus 181.8 +/- 26.7 pg/ml; P < 0.001). Pericardial and plasma atrial natriuretic peptide concentrations showed a significant positive correlation. Pericardial fluid and plasma samples were fractionated using both reverse-phase high-performance liquid chromatography and gel permeation chromatography. Each fraction was assayed for atrial natriuretic peptide by radioimmunoassay, revealing the presence of beta atrial natriuretic peptide as well as alpha- and gamma-atrial natriuretic peptide. 3. The pericardial fluid concentration of cyclic GMP, the intracellular second messenger for atrial natriuretic peptide, was significantly higher in patients with congestive heart failure than in patients without heart failure. PMID- 8403786 TI - Isolation of human cardiac endothelin receptors by a peptide-receptor mobility shift assay. AB - 1. A peptide-protein mobility shift assay has been developed, using native polyacrylamide-gel electrophoresis, that enables the isolation of de-natured receptor proteins from small amounts of human cardiac tissue. 2. Radiolabelled endothelin-1 and related peptides were used to identify and isolate endothelin receptors from partially purified membrane extracts of human atrial tissue. 3. Binding analysis using radiolabelled endothelin-1 gave an equilibrium dissociation constant (Kd) of 2 nmol/l, similar to results from binding experiments conducted directly on tissue. 4. Peptide-receptor complexes were electroeluted from native gels and dissociated. Receptor material was characterized by dot-immunobinding analysis of eluates using an antibody raised against a predicted human endothelin receptor sequence. PMID- 8403787 TI - Magnesium sulphate reverses the carotid vasoconstriction caused by endothelin-I, angiotensin II and neuropeptide-Y, but not that caused by NG-nitro-L-arginine methyl ester, in conscious rats. AB - 1. Magnesium sulphate (MgSO4) has been used for many years in the prevention of eclamptic seizures, but its mechanism of action has never been elucidated. Recent studies suggest that cerebral vasospasm is an important feature of eclampsia and we have developed and tested the hypothesis that MgSO4 can reverse cerebral vasoconstriction. 2. Studies were performed in conscious, male Long Evans rats with pulsed Doppler probes sutured around both common carotid arteries after the external carotid artery had been ligated on the left, thus allowing simultaneous measurement of changes in common and internal carotid blood flow. Intravascular catheters were placed in the abdominal aorta for measurement of systemic blood pressure and in the right jugular vein for administration of drugs. Mean arterial blood pressure and mean Doppler shift signals were used to calculate percentage changes in common and internal carotid vascular conductance. 3. After a period of recovery the animals were infused with endothelin-1, angiotensin II, neuropeptide Y or NG-nitro-L-arginine methyl ester alone or in combination, and MgSO4 in low or high dose was infused when the effects of the vasoconstrictors had become established. 4. MgSO4 itself, at the low dose, had no effect on carotid vascular conductance. Endothelin-1, angiotensin II and neuropeptide-Y all reduced common and internal carotid vascular conductance and this effect was significantly attenuated by low dose MgSO4. The carotid vasoconstrictor action of endothelin-1 was completely abolished by high dose MgSO4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403788 TI - Influence of counterregulatory hormones, independently of hypoglycaemia, on cognitive function, warning symptoms and glucose kinetics. AB - 1. To assess the influence of counterregulatory hormones, independently of neuroglycopaenia, on higher cerebral (cognitive) function, 'hypoglycaemic' warning symptoms and glucose kinetics, 10 healthy subjects participated in two hyperinsulinaemic (2 m-units min-1 kg-1) glucose clamp studies. After 100 min of euglycaemia (plasma glucose level 5 mmol/l), the plasma glucose level was either (a) maintained at 5 mmol/l for 120 min by glucose infusion with concomitant replacement of counterregulatory hormones (continuous infusions of glucagon, adrenaline, noradrenaline, cortisol and growth hormone) to mimic the hormonal milieu normally associated with hypoglycaemia (hormone infusion study) or (b) lowered to 2.8 mmol/l for 120 min (hypoglycaemia study). Assessments were made of cognitive function (P300 auditory evoked responses), symptoms (visual analogue scales) and glucose kinetics (3-[3H]glucose). 2. Hypoglycaemia was associated with an increase in all symptoms (facial flushing, palpitations, tingling, trembling, sweating, hunger, light-headedness and sleepiness, P < 0.01) and all subjects were aware that blood glucose levels had fallen. P300 evoked potential latency increased from 280 +/- 6 to 312 +/- 5 ms (mean +/- SEM, P < 0.01). In contrast, P300 latency and several individual symptoms (hunger, facial flushing, sweating and light-headedness) did not change from baseline during the hormone infusion study (P < 0.05 versus hypoglycaemia). Hepatic glucose production was lower (1.5 +/- 0.4 versus 2.3 +/- 0.3 mg min-1 kg-1, P < 0.05) and peripheral glucose uptake was higher (7.4 +/- 1.0 versus 5.6 +/- 0.6 mg min-1 kg-1, P < 0.01) during infusion of the hormones compared with during hypoglycaemia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403789 TI - Glutathione peroxidase activity and metabolism of arachidonic acid in peripheral blood mononuclear cells from elderly subjects. AB - 1. Since ageing has been associated with a decrease in both immune responses and antioxidant defences, this study was undertaken to compare the glutathione peroxidase activity in peripheral blood mononuclear cells of healthy elderly and young donors. The mean value of glutathione peroxidase activity was significantly lower in the aged group (-36%) than that observed in the young control group (n = 10). 2. This defect was accompanied by a marked increase (+106%) in the oxygenated metabolism of endogenous arachidonic acid by lipoxygenases as judged by the radiolabel associated with hydroxyeicosatetraenoic acids in [3H]arachidonic acid-prelabelled peripheral blood mononuclear cells, whereas the cyclo-oxygenase activity, estimated by the radiolabel associated with thromboxane B2, was not significantly altered. 3. Upon stimulation by the mitogenic lectin concanavalin A, the radioactivity associated with total eicosanoids (free arachidonic acid plus hydroxyeicosatetraenoic acids plus thromboxane B2) was significantly increased over basal levels in the peripheral blood mononuclear cells of both the elderly and the control groups. However, the mitogen-induced increase was lower in the elderly group (+48%) than in the control group (+131%). Upon concanavalin A stimulation, the radioactivity of hydroxyeicosatetraenoic acids was increased by only 96% in peripheral blood mononuclear cells from the elderly group compared with 350% in control cells. Similarly, the radioactivity associated with thromboxane B2 was increased by only 82% in peripheral blood mononuclear cells from the elderly group compared with 218% in control cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403790 TI - Some pitfalls in the measurement of blood glutathione. AB - 1. We report some pitfalls in the measurement of whole blood and plasma glutathione in man. 2. Using a simple tourniquet to the forearm and a 21-gauge needle, blood samples were collected by brachial vein puncture from healthy subjects. Whole blood and plasma were analysed for total glutathione, including the reduced and oxidized forms, by a spectrophotometric recycling method involving the glutathione reductase/NADPH couple. 3. The concentration of oxidized glutathione was determined after treatment of aliquots with either 2 vinylpyridine or N-ethylmaleimide to trap reduced glutathione. Reduced glutathione in the native samples could then be obtained by subtraction. 4. When the reagents were added to separated plasma, 2-vinylpyridine yielded values for oxidized glutathione that were twice as high as with N-ethylmaleimide. In whole blood studies the discrepancy was even greater, and the problem was not resolved by deproteinization of samples with 5-sulphosalicylic acid. Using N ethylmaleimide, levels of oxidized glutathione were less than 1% of total glutathione in whole blood. 5. Despite attempts to minimize haemolysis, lysed erythrocytes contributed on average 25% to the 'plasma' glutathione concentration. PMID- 8403791 TI - Modulation of the anabolic effect of synthetic human parathyroid hormone fragment (I-34) in the bone of growing rats by variations in the dosage regimen. AB - 1. The influence of the time of the day of the administration of synthetic human parathyroid hormone fragment-(1-34) [hPTH-(1-34)] on its anabolic effect in bone was investigated in 23 60-day-old female Wistar rats. Rats were randomly assigned to the groups vehicle control morning, hPTH-(1-34)-treated morning, vehicle control afternoon and hPTH-(1-34)-treated afternoon, and once daily received a subcutaneous injection of 8 micrograms of hPTH-(1-34)/100 g body weight for 11 days. The increase in net intestinal calcium absorption and the increase in calcium balance were not influenced by the time of day of hPTH-(1-34) treatment. Four days after cessation of treatment, the net intestinal calcium absorption and calcium balance in hPTH-(1-34)-treated rats were not different from those of the control rats. 2. Modulation of the anabolic effect by variation of the hPTH-(1 34) dosage regimen was investigated in 43 60-day-old female Wistar rats. Rats were randomly assigned to the groups vehicle control, environmental control, 8 micrograms of hPTH-(1-34)/100 g body weight every 3 days for 24 days, 8 micrograms of hPTH-(1-34)/100 g body weight every 2 days for 16 days, 8 micrograms of hPTH-(1-34)/100 g body weight every day for 8 days, 4 micrograms of hPTH-(1-34)/100 g body weight twice a day for 8 days and 2.7 micrograms of hPTH (1-34)/100 g body weight three times a day for 8 days. In all cases, the total dose of hPTH-(1-34) received was 64 micrograms/100 g body weight.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403792 TI - Comparison between continuous and discrete measurements of breathlessness during exercise in normal subjects using a visual analogue scale. AB - 1. Visual analogue scaling of breathlessness made at discrete intervals during ventilatory stimulation tests can provide useful information about the intensity of this sensation. The aim of the present study was to investigate the use of continuous visual analogue scaling as a means of improving the temporal resolution of this measurement. 2. Six normal naive subjects scaled breathlessness using a visual analogue scale, during steady-state exercise. Further changes in this sensation were induced by either sustained hypercapnia or acute hypoxia; these responses were assessed either continuously or at discrete 30 s intervals and the two scaling methods were compared. 3. The continuous method of assessing breathlessness compared favourably with that of the more established discrete method, providing reproducible measurements in repeated tests equivalent in intensity to those obtained every 30 s. 4. Transient changes in the sensation of breathlessness produced by acute episodes of hypoxia were identified using the continuous scaling method but not with discrete scaling. 5. The continuous method of scaling breathlessness should aid the investigation of the neurophysiological basis of this sensation by allowing temporal relationships between changes in respiratory variables and the sensory consequences to be more carefully defined. PMID- 8403793 TI - Effect of glucocorticosteroid treatment on beta-adrenoceptor subtype function in adipocytes from patients with asthma. AB - 1. Adrenoceptor subtype function was studied in isolated adipocytes obtained by subcutaneous fat biopsies from nine patients with mild asthma. The biopsies were taken before and after 7 days treatment with 25 mg of prednisolone given orally. Lipolytic activity after stimulation with various adrenergic agent was measured, using glycerol release as an index of lipolysis. The number of beta 1- and beta 2 adrenoceptor binding sites was determined in radioligand binding experiments and beta 1- and beta 2-adrenoceptor mRNA levels were measured with a solution hybridization assay. 2. Lipolytic sensitivity (ED50) to isoprenaline, a non selective beta-adrenoceptor agonist, increased 50-fold after treatment (P = 0.04). Sensitivity to terbutaline, a selective beta 2-adrenoceptor agonist, increased 25-fold (P = 0.01), whereas the ED50 values for dobutamine, a selective beta 1-adrenoceptor agonist, did not change significantly. Likewise, the sensitivity to the alpha 2-adrenoceptor agonist, clonidine, and to the drugs acting at post-receptor levels did not change significantly. Basal and maximum lipolytic rates on stimulation were not altered by the treatment. 3. The number of beta 2-adrenoceptor binding sites increased by 60% after treatment (P < 0.05), whereas the beta 1-adrenoceptor binding sites were not affected. The affinity of each receptor subtype for the displacing ligand, ICI 118.551, was not significantly altered by steroids. No significant changes were demonstrated in either beta 1- or beta 2-adrenoceptor mRNA levels. 4. Thus, glucocorticoids selectively increase beta 2-adrenoceptor density and function in patients with asthma, studied by using subcutaneous fat cells as an experimental model. PMID- 8403794 TI - Assessment of adipose tissue metabolism in man: comparison of Fick and microdialysis techniques. PMID- 8403795 TI - Defective enzyme-mediated receptor protection: novel mechanisms in the pathophysiology of hypertension. PMID- 8403796 TI - A possible role for protein kinase C in the regulatory differences between intra abdominal and subcutaneous human adipose tissue. AB - 1. The adenosine A1-receptor agonist N6-(phenylisopropyl)adenosine inhibited lipolysis in a dose-dependent manner in human adipocytes. The effect is mediated by the inhibitory guanine-nucleotide-binding protein(s) that can be phosphorylated and thereby inactivated by protein kinase C. 2. Stimulation of protein kinase C by 12-O-tetradecanoyl-phorbol-13-acetate attenuated the inhibitory effect of the adenosine agonist. 3. Omental fat cells are less sensitive to adenosine than subcutaneous cells, although the stimulatory arm of cyclase regulation appears normal. Protein kinase C activity was measured in the soluble and particulate fractions of human omental and subcutaneous adipose tissue. Omental adipose tissue had a twofold higher membrane-bound and a threefold higher soluble protein kinase C activity. 4. It is therefore possible that the differences in regulation between the two sites are caused by different C kinase activities, causing variable phosphorylation of the inhibitory guanine nucleotide-binding protein(s). PMID- 8403797 TI - Genetic variation associated with differences in the response of plasma apolipoprotein B levels to dietary fibre. AB - 1. We hypothesized that differences within genes whose protein products are involved in apolipoprotein B metabolism could influence the response of plasma apolipoprotein B-containing lipoprotein concentrations to increases in dietary fibre. 2. We studied 67 subjects (43 men and 24 women) who had taken part in parallel 2 week metabolic dietary studies involving either wheat bran or oat bran supplementation. Fasting blood lipid, lipoprotein and apolipoprotein concentrations were measured at the start and end of the 2 week metabolic period. Genotypes were determined using DNA markers for the low-density lipoprotein receptor, apolipoprotein B, apolipoprotein CIII and hepatic lipase gene loci. 3. Reductions in plasma concentrations of apolipoprotein B were significantly different depending on genotype determined with a low-density lipoprotein receptor DNA marker (P = 0.03). There was no significant variation in the reduction of plasma total cholesterol, low-density lipoprotein cholesterol or apolipoprotein B concentrations for alleles of other genes tested. 4. Thus, genetic variability is associated with interindividual differences in the fibre related reduction in plasma apolipoprotein B and apolipoprotein B-containing lipoprotein concentrations. Implementation of current dietary recommendations to reduce plasma lipoprotein levels with fibre may have variable effects in different individuals in part because of structural differences in candidate genes whose products are involved in lipoprotein metabolism. PMID- 8403798 TI - Bombesin reduces food intake after a preload in man by a cholecystokinin independent mechanism. AB - 1. A double-blind study was undertaken to determine whether the infusion of bombesin inhibits the intake of a carbohydrate-rich meal, consumed 15 min after a 300 ml banana shake, in nine lean healthy subjects and whether the possible inhibition of food intake by bombesin is mediated by cholecystokinin. 2. The amount of food eaten during infusion of bombesin (267 +/- 60 g) and bombesin combined with the cholecystokinin-receptor antagonist loxiglumide (269 +/- 39 g) was slightly (P = 0.09) less than during saline infusion (384 +/- 40 g). In addition, preprandial feelings of hunger were significantly less during infusion of both bombesin and bombesin combined with loxiglumide. 3. In conclusion, infusion of bombesin tends to inhibit the intake of a carbohydrate-rich meal after a preload by a cholecystokinin-independent mechanism. PMID- 8403799 TI - Plasma membrane form of phosphatidate phosphohydrolase: a possible role in signal transduction during liver fibrogenesis. AB - 1. Several growth factors important in liver regeneration and fibrosis stimulate phospholipase D in plasma membranes via a receptor/G-protein-coupled mechanism resulting in hydrolysis of phosphatidylcholine to phosphatidate. Phosphatidate can be further hydrolysed to diacylglycerol by phosphatidate phosphohydrolase. Phosphatidate and diacylglycerol can act as 'second-messengers' and regulation of phosphatidate phosphohydrolase activity could control the balance between them. 2. A form of phosphatidate phosphohydrolase, located in the plasma membrane and insensitive to inhibition by N-ethylmaleimide, has recently been identified that is distinct from the 'metabolic' form, which is present in the cytosol and microsomes and is sensitive to N-ethylmaleimide. 3. We have investigated the hypothesis that the balance between regeneration and fibrosis is, in part, determined by the activity of plasma membrane phosphatidate phosphohydrolase through its effect on the phosphatidate/diacylglycerol ratio. N-Ethylmaleimide insensitive and -sensitive phosphatidate phosphohydrolase activities were measured in three hepatic conditions characterized by regeneration and/or fibrosis: alcoholic liver disease in humans (regeneration and fibrosis) and rat livers after either acute CCl-4-induced injury (regeneration) or common bile duct ligation (fibrosis). 4. In patients with alcoholic liver disease, N ethylmaleimide-insensitive phosphatidate phosphohydrolase activity was higher in cirrhotic biopsies (5.82 +/- 0.3 nmol of Pi min-1 mg-1 of protein, n = 19) than in non-cirrhotic biopsies (2.17 +/- 0.2, n = 23) or in wedge biopsies from healthy subjects undergoing routine cholecystectomy (2.16 +/- 0.5, n = 6).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403800 TI - Human pyruvate dehydrogenase complex as an autoantigen in primary biliary cirrhosis. AB - 1. The sera of more than 90% of patients with primary biliary cirrhosis contain antimitochondrial antibodies which react with the E2 component of the pyruvate dehydrogenase complex, identified as the major autoantigen in primary biliary cirrhosis. All previous studies in this area have utilized protein derived from animal tissue or have used recombinant human pyruvate dehydrogenase complex E2 expressed in Escherichia coli. 2. We report the preparation and characterization of native pyruvate dehydrogenase complex and pyruvate dehydrogenase complex E2 from human heart tissue and its application in studies of immune reactivity with the sera of patients with primary biliary cirrhosis. 3. The immune reactivity of sera from patients with primary biliary cirrhosis versus the bovine and human E2/X components of pyruvate dehydrogenase complex was indistinguishable in both immunoblotting and the more sensitive e.l.i.s.a. 4. These findings suggest that the reactivity of sera from patients with primary biliary cirrhosis against the major autoantigen of the disease is a property of that antigen, independent of its human or bovine origin. Furthermore, this justifies the use of bovine pyruvate dehydrogenase complex in past and future work on primary biliary cirrhosis antibody reactivity. PMID- 8403801 TI - Gene expression of metalloproteinases and their inhibitor in renal tissue of New Zealand black/white F1 mice. AB - 1. The present study was carried out to determine how levels of the mRNA of metalloproteinases (metalloproteinase-1, 72 kDa type IV collagenase, metalloproteinase-3 and 92 kDa type IV collagenase) and tissue inhibitor of metalloproteinases are regulated in the renal tissues of New Zealand Black/White F1 mice. 2. mRNA levels for metalloproteinase-1, 72 kDa type IV collagenase, metalloproteinase-3 and tissue inhibitor of metalloproteinases increased significantly with the progression of nephritis in New Zealand Black/White F1 mice. 3. At 48 weeks of age, the levels of mRNA for metalloproteinase-1, 72 kDa type IV collagenase, metalloproteinase-3 and tissue inhibitor of metalloproteinases increased by 8-, 4-, 8- and 15-fold, respectively, in the renal tissues of New Zealand Black/White F1 mice compared with New Zealand White mice. 4. In the kidneys of New Zealand White mice, however, the mRNA levels of these proteins changed little throughout the experimental period. 5. We could not detect expression of mRNA for 9 2 kDa type IV collagenase in the renal tissue of New Zealand Black/White F1 mice at 8 weeks of age or in New Zealand White mice at 8, 24 or 48 weeks of age, whereas we could detect expression of mRNA for this protein in New Zealand Black/White F1 mice at 24 and 48 weeks of age when mononuclear cells had infiltrated the interstitium and surrounding blood vessels. 6. At 24 weeks of age, New Zealand Black/White F1 mice were divided into two groups and received either methylprednisolone or saline injection for 24 weeks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403802 TI - Dissolved urate promotes calcium oxalate crystallization: epitaxy is not the cause. AB - 1. Increasing the concentration of dissolved urate promotes the crystallization of calcium oxalate from urine. The possibility was investigated that this effect may be caused by heterogeneous nucleation of calcium oxalate by particles of crystalline urate. 2. Urine samples were collected from 10 healthy men, centrifuged and filtered, and a solution of sodium urate was added to increase the medium urate concentration from 2.2 to 5.6 mmol/l. Calcium oxalate crystallization was induced by the addition of oxalate, followed by incubation for 90 min in a shaking waterbath at 37 degrees C. The crystalline material was filtered out and the urate concentration was determined in the filtrate. 3. No difference in the urate concentration before and after induction of calcium oxalate crystallization was observed. These findings were confirmed by infra-red spectroscopy, X-ray powder diffraction and ultraviolet wet chemical analysis with detection limits of 5-10, 1.0 and 0.055%, respectively; urate was not detected in the calcium oxalate crystals. 4. In addition, three urine samples were collected and passed through 10 kDa ultrafiltration membranes to remove any colloidal particles which might have been present. The urate concentration was increased and an oxalate load was added as before, prior to incubation at 37 degrees C in a shaking water bath for 5 min, followed by passage through 10 kDa ultrafiltration membranes. Scanning electron microscopy revealed no particles on the membranes thereby indicating that colloidal or crystalline urate was not formed early in the crystallization experiments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403803 TI - Effects of oral and intravenous calcitriol on serum calcium oxalate saturation in dialysis patients. AB - 1. To determine whether the multiple changes in the blood chemistry profile induced by calcitriol may be conducive to secondary systemic oxalosis we have studied nine patients on regular dialysis treatment under three different regimens: (1) oral calcitriol, 0.25 microgram/daily for at least 6 months. (2) off calcitriol, a 1-month withdrawal of the drug, taken as the baseline study period; (3) intravenous calcitriol, 1 microgram three times weekly at the end of dialysis, with tests performed at 1 and 3 months from initiation. 2. Serum concentrations were measured pre- and post-dialysis at the end of each study period. The whole dialysate was used for the determination of the overall calcium and oxalate removal by dialysis. The degree of saturation with calcium oxalate monohydrate was estimated by a computer program. Serum calcitriol concentrations were also assessed. 3. Total and ionized serum calcium did not change on average, although mild hypercalcaemia developed in some patients on intravenous calcitriol. There was an increase in plasma level of oxalate during both oral and intravenous calcitriol treatment, but this was less pronounced during intravenous therapy. Removal of oxalate by dialysis was also greater in patients on oral calcitriol. 4. These increases were probably originated from intestinal absorption and secondary to hyperabsorption of dietary calcium. Consequently, the degree of saturation with calcium oxalate before dialysis rose during calcitriol treatment, irrespective of the route of administration. 5. These results emphasize that, in addition to soft tissue calcification due to calcium phosphates, ectopic calcium oxalate crystallization must also be viewed as a potential risk associated with long-term administration of calcitriol. PMID- 8403804 TI - Ionized and total magnesium levels in cyclosporin-treated renal transplant recipients: relationship with cholesterol and cyclosporin levels. AB - 1. Ionized magnesium, measured using a newly developed ion-selective electrode, total magnesium, and ionized and total calcium were evaluated in 39 stable, long term, cyclosporin-treated renal transplant recipients and compared with those of age-matched, non-transplanted control subjects. Total cholesterol, cyclosporin trough level, serum creatinine, time after-transplant and the ratio of ionized calcium to ionized magnesium were also measured in renal transplant recipients and the relationships between these variables and ionized and total magnesium were evaluated. 2. Renal transplant recipients exhibited marked deficits in ionized magnesium, with a mean value of 0.54 +/- 0.01 mmol/l as compared with 0.61 +/- 0.006 mmol/l for normal control subjects (P < or = 0.05), with a more moderate deficit in total magnesium. Values for ionized and total calcium did not differ. By stepwise linear multiple regression analysis, ionized magnesium was significantly related to cyclosporin trough level and total cholesterol but not to serum creatinine, time after transplant or the dose of cyclosporin. Ionized magnesium correlated inversely with cyclosporin trough level and directly with total cholesterol. The ratio of ionized calcium to ionized magnesium was elevated in renal transplant recipients when compared with control subjects and correlated positively with the cyclosporin trough level. 3. Deficits in ionized magnesium are common during the late post-transplant period in cyclosporin-treated renal transplant recipients. Ionized magnesium may be a more sensitive clinical parameter than total magnesium in this population, in whom total magnesium may be only mildly decreased in the setting of a severe deficit in ionized magnesium. 4. Ionized magnesium correlates with the cyclosporin level.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403805 TI - Potassium channels in hypokalaemic periodic paralysis: a key to the pathogenesis? AB - 1. A possible role for the ATP-sensitive potassium channels in the pathogenesis of hypokalaemic periodic paralysis was investigated. 2. We assessed insulin release and muscle strength after intravenous glucose loading with and without the potassium channel opener pinacidil and the potassium channel blocker glibenclamide in three patients with hypokalaemic periodic paralysis and in a pair of matched control subjects for each patient. 3. A significantly higher initial insulin response (1.5-30 min) was found in the patients with hypokalaemic periodic paralysis in comparison with the control subjects. During potassium channel blocking with glibenclamide the insulin release was more enhanced in patients than in control subjects. On the other hand, the potassium channel opener pinacidil impaired the insulin release in healthy control subjects but not in patients. The serum glucose levels showed no differences between patients and control subjects. In one of the patients with hypokalaemic periodic paralysis glucose loading resulted in a fall in muscle strength, which did not occur during the administration of pinacidil. 4. These findings suggest a disturbance in the ATP-sensitive potassium channel in patients with hypokalaemic periodic paralysis, which is not limited to pancreatic beta cells, but may be also involved in the abnormal behaviour of skeletal muscle. PMID- 8403806 TI - Relationship between insulin release, antinatriuresis and hypokalaemia after glucose ingestion in normal and hypertensive man. AB - 1. Insulin simultaneously causes hypokalaemia and antinatriuresis, and it has been suggested that the two effects are tightly coupled. Whether these actions are preserved in patients with essential hypertension is not known. 2. Eight hypertensive patients and eight normotensive control subjects were studied before and after the ingestion of 75 g of glucose. Despite similar glycaemic profiles, the patients showed a hyperinsulinaemic response incremental area 49 +/- 8 versus 27 +/- 6 nmol l-1 3 h, P < 0.04) but a blunted hypokalaemic response (-7 +/- 1 versus -16 +/- 1%, P < 0.001). Both absolute and fractional urinary excretion of sodium and potassium were significantly decreased during glucose-induced hyperinsulinaemia in hypertensive patients as well as in normotensive subjects (P < 0.05 for all changes). 3. To test whether hypokalaemia is required for insulin induced antinatriuresis, each hypertensive patient received another oral glucose load during which enough potassium chloride was given to clamp the plasma potassium concentration at baseline. Under these conditions, significant insulin induced antinatriuresis still occurred. In addition, whereas the glycaemic profile was superimposable, the response of the plasma insulin concentration was significantly greater with than without maintenance of the plasma potassium concentration (total area 79 +/- 14 versus 63 +/- 8 nmol l-1 3 h, P < 0.04). 4. We conclude that (a) insulin causes antinatriuresis, antikaliuresis and hypokalaemia under physiological conditions; (b) in hyperinsulinaemic (insulin resistant) patients with essential hypertension, the antinatriuretic action of insulin is quantitatively preserved; and (c) clamping plasma potassium levels prevents insulin-induced antikaliuresis but not antinatriuresis, and potentiates the insulin secretory response to glucose. PMID- 8403807 TI - 1H-nuclear magnetic resonance studies of human synovial fluid in arthritic disease states as an aid to confirming metabolic activity in the synovial cavity. AB - 1. A 1H-n.m.r. method was used to measure concentrations of valine, alanine, lactate, acetate, hyaluronan and lipids in synovial fluid obtained, during the normal course of examination from the knee joints of patients attending rheumatology and orthopaedic clinics. Fluid was available from 16 patients with osteoarthritis, 18 patients with rheumatoid arthritis, four patients with meniscal tear and one patient each with systemic lupus erythematosis, mono arthritis, synovitis and loose bodies. Four normal specimens were obtained for comparison. 2. Valine, alanine and acetate levels all showed a normal Gaussian distribution, reflecting the distributions within the serum of the sample population. 3. Lactate concentrations divided into two distinct patterns. At concentrations below 2.5 mmol/l the lactate levels showed a Gaussian distribution, reflecting the distribution in normal serum. The normal synovial fluid specimens belong to this distribution. Above 2.5-3.0 mmol/l, lactate levels were asymmetric in distribution with a long tail at higher concentrations. These high levels of lactate can be explained by the generation of lactate through anaerobic metabolism within the synovial cavity. This metabolic process is triggered by a general inflammatory condition such as in rheumatoid arthritis. 4. The distribution of n.m.r.-observable lipid concentrations in rheumatoid arthritis and osteoarthritis each shows a normal distribution and the mean concentration is significantly higher in rheumatoid arthritis. 5. An increased n.m.r.-observable hyaluronan concentration is associated with an inflammatory situation. 6. It is concluded that raised levels of lactate and n.m.r.-observable hyaluronan and lipids are useful markers to aid the clinical distinction between rheumatoid arthritis and osteoarthritis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403808 TI - Elevated serum superoxide dismutase levels correlate with disease severity and neutrophil degranulation in idiopathic pulmonary fibrosis. AB - 1. Tissue damage in idiopathic pulmonary fibrosis is due in part to oxidant antioxidant imbalance. 2. We evaluated the serum levels of the antioxidant enzyme Cu/Zn superoxide dismutase (EC 1.15.1.1) in 25 patients with idiopathic pulmonary fibrosis, 34 patients with sarcoidosis and 40 healthy control subjects by an enzyme immunometric assay. 3. We found that patients with idiopathic pulmonary fibrosis have higher serum Cu/Zn superoxide dismutase levels than control subjects and patients with sarcoidosis. In addition, serum Cu/Zn superoxide dismutase levels correlate with disease severity indexes in patients with idiopathic pulmonary fibrosis. 4. The increase in serum Cu/Zn superoxide dismutase level in idiopathic pulmonary fibrosis could depend on degranulation of activated neutrophils or release from damaged cells. To elucidate the contribution of neutrophil degranulation we determined the polymorphonuclear cell elastase level in the same specimens. We found a strong correlation between serum Cu/Zn superoxide dismutase and polymorphonuclear cell elastase activities, and, in patients with idiopathic pulmonary fibrosis, we observed higher levels of polymorphonuclear cell elastase than in control subjects and patients with sarcoidosis, which correlated positively with disease severity indexes. 5. Cu/Zn superoxide dismutase can catalyse the dismutation of O2 into H2O2 and generate OH.. These oxygen radicals are probably the major factors responsible for tissue damage (in particular, alveolar and endothelial cells) and fibrosis in experimental lung injury.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403809 TI - Vasodilatory action of the calcium antagonist amlodipine on large and resistance pulmonary arteries from normoxic and chronically hypoxic rats. AB - 1. Isolated rat aorta and pulmonary arteries were maximally precontracted with 100 mmol/l KCl, and the vasorelaxation due to the dihydropyridine calcium antagonist amlodipine was measured. The response of large pulmonary arteries (mean lumen diameter 983 microns) was directly compared with that of isolated pulmonary resistance vessels (mean lumen diameter 259 microns) from both normoxic animals and animals exposed to chronic hypoxia. 2. Amlodipine caused a significant relaxation of aorta (P < 0.001). A significant relaxation of large and resistance pulmonary arteries from both normoxic and chronically hypoxic animals was also demonstrated at all doses tested (P < 0.05) or less). 3. Amlodipine produced significantly more relaxation in pulmonary resistance vessels than in large pulmonary arteries from both normoxic and chronically hypoxic rats (P < 0.02). 4. The action of amlodipine was slow in onset and persistent in all vessels studied. In the pulmonary vessels from normoxic animals both the rate of onset and the magnitude of effect was proportional to the drug concentration (P < 0.001). 5. These results demonstrate that amlodipine is a potent inhibitor of KCl induced contractions in rat pulmonary arteries with a preferential action in pulmonary resistance vessels. PMID- 8403810 TI - Validity of peak expiratory flow rate variability for the diagnosis of asthma. AB - 1. The aim of this study was to determine the validity of various numerical indices of peak flow variability as tests for asthma. 2. Normal (n = 24) and asthmatic (n = 123) subjects aged 10-70 years were recruited from the community. Asthma was diagnosed by full clinical assessment and was active during the previous year. Subjects recorded their peak flow immediately after rising in the morning, at 18.00 hours and just before retiring at night for 12 days. 3. The most discriminating index was the highest peak flow variability which occurred within any 1 day during the 12-day recording period, calculated as the difference between the maximum and minimum peak flows expressed as a percentage of the minimum peak flow on that day (the proposed index). This index was < 20% (90th centile) in 96% of the normal subjects and > or = 20% in 89% of the asthmatic subjects. 4. Standard indices of mean peak flow, the forced expiratory volume in 1.0 s and its responsiveness to salbutamol, had much lower sensitivities than peak flow variability. 5. It is concluded that numerical indices of peak flow variability are highly valid tests for asthma. Using an upper limit of normal of 20%, the proposed index of peak flow variability discriminates better than other indices between asthmatic and normal subjects. PMID- 8403811 TI - Preliminary study analysis suggests similar efficacy for didanosine and zalcitabine. PMID- 8403812 TI - Ticlopidine-induced cholestatic jaundice. PMID- 8403813 TI - Pharmacologic and clinical considerations in selecting crystalloid, colloidal, and oxygen-carrying resuscitation fluids, Part 2. PMID- 8403814 TI - Pharmacologic management of adult idiopathic nephrotic syndrome. AB - The pathophysiology, clinical features, complications, and pharmacologic management of adult idiopathic nephrotic syndrome are reviewed. Loss of plasma proteins in the urine is the primary process leading to the nephrotic syndrome, which is characterized by hypoalbuminemia, hyperlipidemia, and edema. The four principal causes, or subclasses, of adult idiopathic nephrotic syndrome are membranous nephropathy (MN), minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), and membranoproliferative glomerulonephritis (MPGN); definitive diagnosis requires histologic examination of a renal biopsy specimen. Treatment of nephrotic syndrome may be directed at the specific cause of the proteinuria, the proteinuria itself, or the complications induced by the syndrome. The four subclasses of nephrotic syndrome vary in their response to therapy. Corticosteroids, alone or in combination with cytotoxic agents, and cyclosporine have been used to induce partial or complete remission in patients with MN, MCD, and FSGS; combinations of corticosteroids, cytotoxic agents, platelet inhibitors, and anticoagulants have been used to treat patients with MPGN. Treatment of proteinuria involves dietary protein restriction with the possible addition of an angiotensin-converting-enzyme inhibitor or a nonsteroidal anti-inflammatory drug. Management of the complications of nephrotic syndrome encompasses the use of diuretics; a low-cholesterol, low-fat diet; lipid-lowering agents; and anticoagulants. Patients with nephrotic syndrome are in a constant state of flux with respect to fluid status, organ function, and critical protein balance. Treatment is based on the histologic subclass of the disease. PMID- 8403815 TI - Drug-induced pancreatitis. AB - Recent information about drugs implicated in causing pancreatitis is summarized. Although the frequency of drug-induced acute pancreatitis is generally low, the disease is associated with substantial morbidity and mortality, which makes timely identification of the offending agent important. Mechanisms suggested for drug-induced pancreatitis include pancreatic duct constriction; immunosuppression; cytotoxic, osmotic, pressure, or metabolic effects; arteriolar thrombosis; direct cellular toxicity; and hepatic involvement. Agents reported to have a definite association with pancreatitis are asparaginase, azathioprine, didanosine, estrogens, furosemide, mercaptopurine, pentamidine, sulfonamides, sulindac, tetracyclines, thiazides, and valproic acid. Agents reported to have a probable association with pancreatitis include cimetidine, clozapine, corticosteroids, endoscopic retrograde cholangiopancreatography contrast media, methyldopa, metronidazole, salicylates, and zalcitabine. Agents reported to have a questionable association with pancreatitis include acetaminophen, cyclosporine, cytarabine, erythromycin and roxithromycin, ketoprofen, metolazone, and octreotide. When ethanol abuse and biliary disease are ruled out as etiologies for pancreatitis, the possibility of drug-induced disease should be investigated. PMID- 8403816 TI - Amorous, disinhibited behavior associated with propofol. PMID- 8403817 TI - Criteria for use of ofloxacin and lomefloxacin in adult inpatients and outpatients. PMID- 8403818 TI - Co-ordinating diabetes services in a health district. PMID- 8403819 TI - Potential therapeutic approaches to the treatment or prevention of diabetic neuropathy: evidence from experimental studies. AB - Recent investigations using experimental models of diabetes mellitus have emphasized the importance of impaired blood flow for the development of nerve dysfunction. Other observations suggest that this may also be the case for patients. A number of studies have revealed that several types of vasodilators can prevent or successfully treat early conduction abnormalities in diabetic rodents. These include alpha 1-adrenoreceptor antagonists, calcium channel blockers, agents that inhibit the renin-angiotensin system, and vasomodulator prostanoids. Other treatments applied to animal models, such as omega-6 essential fatty acids, aldose reductase inhibitors, aminoguanidine which prevents the formation of advanced glycation end-products, and anti-oxidants all appear to have vascular-related effects that lead to improvements in nerve conduction. These findings suggest that endothelial dysfunction and oxidative stress could be important factors in the aetiology of diabetic neuropathy. Studies have also focused on deficits in axon growth and regeneration, their relation to impaired neuronal synthesis and transport of growth-related chemicals, and neuronotrophic abnormalities. Taken together, the data give rise to the notion that an optimal therapeutic strategy could consist of improving the microenvironment of damaged nerve fibres by manipulating nerve blood flow while concurrently encouraging repair with trophic agents. PMID- 8403820 TI - Difficulties in classifying diabetes at presentation in the young adult. AB - All newly diagnosed diabetic patients in Sweden aged 15-34 years have been registered since 1983. In this study the clinical characteristics initially and after 2.5-3 years were evaluated by a questionnaire to the patient's physician and by non-fasting C-peptide. The study comprised patients registered 1983-84, and for 281 patients (37%), complete information was obtained. At diagnosis 75% were classified as Type 1, 19% as Type 2, and 6% as secondary diabetes or as uncertain by their physician. Twenty patients (7.1%) were reported to have ketoacidosis. Seventy-five percent were treated with insulin, 7% with oral hypoglycaemic agents (OHG), and 18% with diet alone. At follow-up 71% were classified as Type 1, 21% as Type 2, and 8% as secondary or uncertain while treatment was 82% insulin, 8% OHG, and 9% diet. During the follow-up period 42% of the initially non-insulin-treated patients were put on insulin whereas only a few stopped insulin treatment. Patients treated with diet or OHG at follow-up were older, had higher percent desirable weight, and lower blood glucose at diagnosis than patients treated with insulin. All except one patient had measurable random C-peptide at follow-up and mean values were for patients treated with insulin 0.55, OHG 1.41 and diet alone 1.29 nmol l-1. Random blood glucose results were similar. In conclusion the majority of newly diagnosed patients in the age group 15-34 years have the characteristics of Type 1 diabetes and Type 2 diabetes is rare before 25-30 years of age.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403821 TI - Sensori-motor function in older persons with diabetes. AB - Twenty-five persons with diabetes (aged 55-83 years) who were living independently in the community, and 40 age- and sex-matched non-diabetic controls were assessed for tactile sensitivity, vibration sense, proprioception, quadriceps strength and body sway. In both men and women, those with diabetes performed significantly worse in tests of body sway on firm and compliant surfaces compared with the control subjects after controlling for weight and body mass index. The female diabetic subjects also performed significantly worse in tests of peripheral sensation and strength compared with controls. Age-related declines in sensori-motor function were greater in the diabetic group (r = 0.55 0.75) than in the controls (r < 0.44), while within the diabetic group, duration of diabetes and vibration sense were significantly correlated with sway on a compliant (foam rubber) surface with the eyes open (partial r = 0.52, p < 0.01 and r = 0.55, p < 0.01, respectively). The study findings provide evidence that older people with diabetes have problems with stability and related sensori-motor factors which may place them at increased risk of falls. PMID- 8403822 TI - The performance of three measures of health status in an outpatient diabetes population. AB - Both the late complications of diabetes and the means used to prevent them have a significant impact on the lives of people with the condition. Measuring quality of life is therefore important in assessing clinical need and evaluating the success of management. Three approaches to measuring health status were therefore compared in 284 randomly selected out-patients attending a hospital diabetes service. The measures used were the Nottingham Health Profile (NHP), four categories of an anglicized version of the Sickness Impact Profile (the Functional Limitations Profile (FLP)), and a scale of Positive Well-being (PWB). The results were found to be independent of questionnaire order and place of completion. The distributions of scores on the NHP and FLP scales were highly skewed, with a majority of cases scoring zero. NHP and FLP scores were related (p < 0.001) to age, but not otherwise to type of diabetes. Patients with angina, circulatory problems, and neuropathy scored significantly higher (up to p < 0.001) on several dimensions/categories of the NHP and FLP, but not the PWB scale. Severe visual impairment (worse than 6/36) was only related (p < 0.005) to NHP 'Mobility' and FLP 'Ambulation' and 'Home management'. Validation of scores by interview gave satisfactory results on all dimensions of the NHP except 'Energy', and all FLP categories except 'Recreations and pastimes'. No statistically significant association was observed between the PWB and the interviewer's assessments, but it did correlate (up to tau = 0.45, p < 0.001) with some social and psychological dimensions/categories of the NHP/FLP. In conclusion the PWB scale is independent of physical disability.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403823 TI - Mortality among type 2 diabetic individuals and associated risk factors: the Three City Study. AB - The 10 year mortality experience was determined in a population-based cohort of 540 Type 2 diabetic individuals. The association between potential risk factors and all causes mortality was examined. Diabetes was not mentioned anywhere on the death certificate in 46% of 274 decedents. Diseases of the circulatory system (ICD9-390-459) accounted for the majority (62%) of deaths in this cohort. Ten year survival was poorer than expected for both men and women compared to the age and sex-matched Minnesota population. Standardized mortality ratios for selected causes of death indicated excess for cardiovascular disease (ICD9-390-459), coronary heart disease (ICD9 410-414) and cerebrovascular disease. Baseline variables associated with all causes of mortality included age and a history of macrovascular disease. These findings indicate that mortality data significantly underestimate the magnitude of diabetes and that individuals with diabetes have poorer survival than non-diabetic individuals. PMID- 8403824 TI - The acute effect of preprandial exogenous and endogenous sulphonylurea-stimulated insulin secretion on postprandial glucose excursions in patients with type 2 diabetes. AB - Sulphonylureas improve glucose tolerance by stimulating insulin secretion. Whether improved glucose tolerance results from enhanced early insulin release or greater total insulin secretion is not clear. Insulin response to a test meal in Type 2 diabetic subjects with and without a single dose (2.5 mg) of oral and intravenous glipizide were, therefore, measured. Intravenous glipizide enhanced early insulin release more than oral glipizide (134% and 80% vs control; p < 0.01), whereas total insulin release was equally improved (78% and 54% vs control; p < 0.01). Despite slight differences in insulin release, there was no difference in glucose tolerance (median area under concentration curve (AUC); 66.6 vs 61.9 mmol x min l-1; NS). The test meal was repeated after a bolus of intravenous insulin at the beginning of the meal. This allowed comparison of the effect of exogenous and endogenous insulin supply on postprandial glucose excursions. In spite of an early and fivefold larger rise in serum insulin after intravenous administration of the hormone than after intravenous glipizide (725% vs 134%; p < 0.01), postprandial glucose was no better than after glipizide (median AUC; 87.8 vs 66.6 mmol x min l-1; NS). In contrast, glucose tolerance was better after oral glipizide compared to intravenous insulin (median AUC; 61.9 vs 87.8 mmol x min l-1; p < 0.05). In conclusion, the total amount of insulin secreted seems more important than the timing of the insulin release for the postprandial glucose tolerance in Type 2 diabetic subjects. Neither endogenous nor peripheral premeal supply of insulin could normalize postprandial glucose excursions in patients with Type 2 diabetes. PMID- 8403825 TI - The relationship between plasminogen activator inhibitor-1 and insulin resistance in newly diagnosed type 2 diabetes mellitus. AB - It is not clear whether elevated levels of the fibrinolytic inhibitor, plasminogen activator inhibitor-1 (PAl-1) in Type 2 diabetes mellitus are the result of obesity or coexistent atherosclerosis. Therefore the relationship between PAl-1 and insulin resistance, determined by the homeostasis model assessment (HOMA) was investigated in a group of 26 insulin-resistant, normotensive newly diagnosed Type 2 diabetic patients with a low probability of atherosclerosis. Compared with a normal control group, closely matched for body mass index (BMI), fibrinolytic activity was depressed in the diabetic patients due to elevated levels of the inhibitor PAl-1, 17.6 (11.1-28) vs 8.4 (4.9-14.1) IU ml-1, p < 0.001. PAl-1 was related to BMI, r = 0.59, p < 0.001 plasma insulin, r = 0.66, p < 0.001; insulin resistance, r = 0.54, p < 0.005 and urinary albumin excretion, r = 0.48, p < 0.01, but not HbA1c or fasting glucose. PAl-1 was not related to blood pressure or plasma triglyceride levels. This study suggests that at the time of diagnosis of Type 2 diabetes mellitus, elevated PAl-1 levels are already linked to other risk factors for vascular disease including hyperinsulinaemia, insulin resistance, and urinary albumin excretion, and this is not the result of obesity or coexistent atherosclerosis. PMID- 8403826 TI - Comparison of platelet aggregability in Japanese type 2 diabetic patients with and without microalbuminuria. AB - Microalbuminuria is associated with higher cardiovascular morbidity and mortality in Type 2 (non-insulin-dependent) diabetic patients. This study was designed to assess whether Type 2 diabetic patients with microalbuminuria (urinary albumin excretion rate (AER) 20-200 microgram min-1) is associated with alterations in platelet aggregability as compared with those with normal urinary albumin excretion (AER < 20 microgram min-1). Platelet aggregability was compared between 21 Japanese Type 2 diabetic patients with microalbuminuria and 21 individually pair-matched (for age, sex, body mass index, treatment, and HbA1c level) patients with normoalbuminuria. The in vitro platelet aggregation induced by 1.0 and 3.0 mumol l-1 ADP and 0.5 and 1.0 mg l-1 collagen was measured using platelet-rich plasma. No significant differences were observed between the two groups in the values for maximum percent platelet aggregation, percent aggregation at 3 min, and aggregation velocities after adding ADP or collagen. Microalbuminuric patients had significantly higher mean values for systolic (p < 0.004) and diastolic (p < 0.02) blood pressures and plasma fibrinogen level (p < 0.03) as compared with the respective mean values in normoalbuminuric patients. The results suggest that Japanese microalbuminuric Type 2 diabetic patients do not differ in the degree of platelet aggregability as compared with normoalbuminuric patients, despite an increase in certain other coronary risk factors. PMID- 8403827 TI - Altered regulation of cholesterol metabolism in type I diabetic women during the menstrual cycle. AB - This study examines the relationship of cellular cholesterol metabolism to oestrogen and progesterone during the menstrual cycle in diabetic and non diabetic subjects. Nine premenopausal diabetic women were compared to nine non diabetic women of the same age. Oestrogen, progesterone, lipoproteins, including lipoprotein (a) (Lp(a)) and cholesteryl ester transfer protein (CETP) were determined in serum. Cellular cholesterol content and cellular cholesterol synthesis were measured in mononuclear leucocytes. There was no significant change in serum lipoproteins including Lp(a) during the cycle in either group. CETP activity was significantly higher over the 4 weeks in the diabetic patients compared with non-diabetic subjects (mean 463 +/- 30 mumol l-1 h-1 vs 405 +/- 28 mumol l-1 h-1, p < 0.01). Serum high density lipoprotein (HDL) cholesterol was significantly lower during the 4 weeks in the diabetic patients (1.7 +/- 0.1 mmol l-1 vs 1.8 +/- 0.1 mmol-1, p < 0.05). Cellular cholesterol synthesis decreased steadily up to the third week in cells from the control subjects whereas there was no significant change in cells from diabetic patients whose cellular cholesterol synthesis was higher at week 3 compared with non-diabetic subjects (663 +/- 54 nmol mg-1 cell protein vs 432 +/- 43 nmol mg-1 cell protein, two-way interaction p < 0.05). There was a significant negative correlation between cellular cholesterol synthesis and serum oestrogen in the non-diabetic subjects (p < 0.05) but not in the diabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403828 TI - Glycaemic responses in type 2 diabetic patients to various mixed meals taken at home. AB - Eight Type 2 diabetic patients ate and prepared five different meals at home, taking each meal on two separate occasions. They measured their blood glucose just before eating and 30, 60, 120, and 180 min after the meal. The meals varied in energy and dietary fibre content and in the ratio (by energy) of carbohydrate to fat. Total energy content of the meals had little effect on the postprandial glycaemic responses nor were the responses reduced by meals with high dietary fibre content. The ratio of carbohydrate to fat did not significantly affect postprandial glycaemic responses when meals were low in fibre. However, postprandial glycaemic responses were significantly greater in the meal with a high ratio of carbohydrate to fat, high in fibre and low in energy compared with those after the equicaloric meal low in carbohydrate to fat ratio and low in fibre (area under the curve 683 +/- 131 vs 306 +/- 55 mmol l-1 min-1, p < 0.05). Fat intake of 35% of energy may be compatible with improved postprandial blood glucose concentrations. Many meal combinations need to be studied in order to provide reliable information for diabetic patients. The method outlined proved producible (within patient coefficient of variation 13%), easy to perform and inexpensive. PMID- 8403829 TI - Hypoglycaemia increases the gastric emptying rate in patients with type 1 diabetes mellitus. AB - The effect of insulin-induced hypoglycaemia on the gastric emptying rate was studied in eight patients with Type 1 diabetes mellitus of short duration (1-5 years). Gastric emptying was studied using a scintigraphic technique. All patients were studied twice, both during euglycaemia and insulin-induced hypoglycaemia. The blood glucose concentration was adjusted with an insulin glucose clamp technique. All patients were examined in a standardized way, undergoing the first examination during euglycaemia, with a blood glucose concentration of 4-7 mmol l-1, and the second during hypoglycaemia, with a mean blood glucose concentration of 1.9 +/- 0.3 mmol l-1 at the time of starting the gastric emptying study. During hypoglycaemia both the liquid and the solid gastric emptying rates were significantly increased compared to the rate during euglycaemia. The time to empty 50% of the radioactivity from the stomach for liquid was 48.5 +/- 9.8 min during euglycaemia, compared to 27.6 +/- 20.2 min during hypoglycaemia, p < 0.001. The time to empty 50% of the radioactivity from the stomach for solid food was 48.7 +/- 10.3 min and 23.2 +/- 15.9 min, respectively, p < 0.001. In conclusion, it appears that insulin-induced hypoglycaemia increases the gastric emptying rate in patients with Type 1 diabetes mellitus. PMID- 8403830 TI - Autonomic and peripheral nerve function in adolescents with and without diabetes. AB - In this study reference ranges were established for autonomic and peripheral nerve tests in 122 non-diabetic adolescents. Regression analysis was used to evaluate the effect of age and gender on neurological function. Increasing age was associated with: less heart rate variability during deep breathing (p = 0.03), higher thermal threshold for cold at the wrist (p = 0.009), and higher vibration threshold at the toe (p = 0.001) and medial malleolus (p = 0.01). Male gender was associated with higher Valsalva ratio (p = 0.0004), higher thermal threshold for hot at the foot (p = 0.002), and higher vibration threshold at the malleolus (p = 0.03). The REFVAL programme was used to determine parametric or non-parametric reference limits: the 5% limits for autonomic and 95% limits for peripheral tests. One hundred and eighty-one adolescents with diabetes were studied under identical conditions and similar effects of age and gender were found. Twenty-eight percent of the group with diabetes had at least one abnormal autonomic test result out of four (expected 18.5%); 24% had at least one abnormal peripheral test result out of six (expected 26.5%). Glycaemic control was associated with autonomic (p = 0.04) but not peripheral abnormalities. Using multiple regression analysis and adjusting for age and gender, there was no effect of diabetes duration or glycaemic control on neurological function. PMID- 8403831 TI - The effectiveness of a primary-care-based diabetes education service. AB - The efficacy of structured education for 158 Type 2 diabetic patients in primary care (80 male, mean age 63 yr, median diabetes duration 3 yr) was assessed with respect to change in knowledge of diabetes, weight, and haemoglobin A1 over a 6 month period. The programme supplemented a primary care initiative in our semi rural population. Teaching was carried out by a Diabetes Nurse Educator within primary care health centres (141 patients) and a hospital diabetes clinic (17 patients). For all patients mean baseline questionnaire score (maximum possible 12) was 6.2 rising after the programme to 10.5 (p < 0.01). At 6 months mean score fell to 9.5 (p < 0.01 compared to end of the programme), but still significantly better than baseline (p < 0.01). For patients on the primary-care-based programme mean haemoglobin A1 at baseline was 10.7% (normal range 6%-9%) decreasing after 6 months to 9.6% (p < 0.01). No significant changes were found in mean weight. Unlike many previous studies, these results demonstrate a highly beneficial effect not only on knowledge but also on metabolic control in patients who received their education in the primary-care setting. These results have obvious implications for patients residing in rural or semi-rural populations. PMID- 8403832 TI - The description and classification of diabetic foot lesions. AB - Diabetic foot ulcers are of many types and different ulcers require management in different ways. Their optimal management is currently hindered by lack of a useful working classification. Such a classification must be flexible enough to be applied to all lesions likely to be encountered but specific enough to enable clear definition of an individual lesion. It must also be simple enough to ensure that it is understood by all categories of health care workers, whether specialist or not. An attempt has been made to devise a classification--based on the key elements used in describing foot lesions--and it is put forward to act as stimulus for debate. It is based on the clinical definition of infection, ischaemia, and neuropathy. Although two of these, or even all three, may be found in the same foot, they should be considered in the order given because this reflects the sequence of clinical decisions which should be made. The adoption of a classification such as this would aid education, communication, research and audit, and would lead to better management of ulcers. PMID- 8403833 TI - The mental model theory of conditional reasoning: critical appraisal and revision. AB - Johnson-Laird and Byrne (1991) present a theory of conditional inference based upon the manipulation of mental models. In the present paper, the theory is critically examined with regard to its ability to account for psychological data, principally with respect to the rate at which people draw the four basic inferences of modus ponens (MP), denial of the antecedent (DA), affirmation of the consequent (AC) and modus tollens (MT). It is argued first that the theory is unclear in its definition and in particular with regard to predictions of problem difficulty. Clarification and specification of principles are consequently provided here. Next, it is argued that there are a number of phenomena in the conditional reasoning literature for which the theory cannot account in its present form. Specifically, (a) the relatively frequency of DA and AC inferences on affirmative conditional is not as predicted by the theory, (b) differences occur between inferences on if then and only if rules beyond the capacity of the theory to explain and (c) there is no account of the "negative conclusion bias" observed when negated components are introduced into the rules. A number of revisions to the mental model theory of conditional reasoning are proposed in order to account for these findings. PMID- 8403834 TI - From rote learning to system building: acquiring verb morphology in children and connectionist nets. AB - The traditional account of the acquisition of English verb morphology supposes that a dual architecture underlies the transition from early rote-learning processes (in which past tense forms of verbs are correctly produced) to the systematic treatment of verbs (in which irregular verbs are prone to error). A connectionist account supposes that this transition can occur in a single mechanism (in the form of a neural network) driven by gradual quantitative changes in the size of the training set to which the network is exposed. In this paper, a series of simulations is reported in which a multi-layered perceptron learns to map verb stems to past tense forms analogous to the mappings found in the English past tense system. By expanding the training set in a gradual, incremental fashion and evaluating network performance on both trained and novel verbs at successive points in learning, it is demonstrated that the network undergoes reorganizations that result in a shift from a mode of rote learning to a systematic treatment of verbs. Furthermore, we show that this reorganizational transition is dependent upon the number of regular and irregular verbs in the training set and is sensitive to the phonological sub-regularities characterizing the irregular verbs. The pattern of errors observed is compared to that of children acquiring the English past tense, as well as children's performance on experimental studies with nonsense verbs. It is concluded that a connectionist approach offers a viable alternative account of the acquisition of English verb morphology, given the current state of empirical evidence relating to processes of acquisition in young children. PMID- 8403835 TI - Learning and development in neural networks: the importance of starting small. AB - It is a striking fact that in humans the greatest learning occurs precisely at that point in time--childhood--when the most dramatic maturational changes also occur. This report describes possible synergistic interactions between maturational change and the ability to learn a complex domain (language), as investigated in connectionist networks. The networks are trained to process complex sentences involving relative clauses, number agreement, and several types of verb argument structure. Training fails in the case of networks which are fully formed and 'adultlike' in their capacity. Training succeeds only when networks begin with limited working memory and gradually 'mature' to the adult state. This result suggests that rather than being a limitation, developmental restrictions on resources may constitute a necessary prerequisite for mastering certain complex domains. Specifically, successful learning may depend on starting small. PMID- 8403836 TI - Dermatophilus congolensis infection (Dermatophilosis) in animals and man! An update. AB - Dermatophilus congolensis infection (dermatophilosis) is an acute, subacute or chronic skin disease affecting a wide range of species of animals and man. It is world widely distributed but more prevalent in the humid, tropics and subtropics. The disease is now being reviewed in detail. In the present review, the geographical distribution, history and nomenclature, animal hosts affected, aetiological agent and clinical signs of the disease are discussed extensively. Emphasis is also given on the predisposing factors such as rainfall, humidity, ectoparasites etc. under epizootiology. Pathogenesis, pathology, immunology treatment, control and prophylaxis are other areas well covered. The economic importance of the disease is also stressed and a new approach (biological approach) to treatment and control of the disease is being described in this review. It was concluded that in view of the importance of the disease for the increase of livestock and leather production in the tropical and subtropical region especially in Africa, an international centre for dermatophilosis research is highly needed. PMID- 8403837 TI - Spotted fever group rickettsial infection in south-eastern Australia: isolation of rickettsiae. AB - Flinders Island spotted fever (FISF), a spotted fever group (SFG) rickettsial disease first described in 1991, occurs in south-eastern Australia. The isolation of the aetiological agent is described for the first time having been obtained from the blood of two patients. An additional 22 cases are also reported. Of these patients four had positive initial serology, and 20 showed seroconversion (using Rickettsia australis as antigen). Acute phase blood specimens taken from seven patients caused neonatal mice to seroconvert to R. australis and a blood specimen from one of these patients (and one other) yielded rickettsiae. A field survey for possible reservoir and vector animals on Flinders Island, Tasmania and in Gippsland, Victoria (both in south-eastern Australia) yielded 217 vertebrates and 1445 invertebrate ectoparasites, mostly ticks. Ixodes cornuatus from humans and dogs in Gippsland produced seroconversion to SFG rickettsia when inoculated into mice but no invertebrate pools from Flinders Island produced seroconversion in mice. Haemolymph from an individual I. cornuatus removed from a human in Gippsland, yielded a SFG rickettsia on tissue culture. Sera from several species of native vertebrates, especially the bush rat, Rattus fuscipes, were positive for antibodies to SFG rickettsia. PMID- 8403838 TI - Field evaluation of the efficacy of Romovac 50, a new inactivated, adjuvanted bovine rotavirus vaccine. AB - An inactivated bovine rotavirus vaccine, prepared with an adjuvant which gives a water-in-oil emulsion (Montanide ISA 50, adjuvant) was used in field trials for prevention of calf neonatal diarrhoea. The vaccine, which was designated as Romovac 50, when injected into cows in their last month of pregnancy, proved to be as effective as the traditional vaccine prepared with Freund's incomplete adjuvant. Thus, the incidence of the naturally occurring neonatal diarrhoea was significantly reduced in calves which were fed mammary secretion from their vaccinated dams, compared to the calves delivered from the unvaccinated cows. Romovac 50 also has the advantages over the traditional Freund's adjuvanted vaccine in that it is less viscous and thus more readily injected and less irritant to tissues. PMID- 8403839 TI - Seroprevalence of infectious bursal disease (IBD) in parts of Tamil Nadu, India. AB - Sera samples of 400 apparently healthy broiler poultry from 23 commercial farms and 120 sera samples from commercial layer poultry [with specific lesions of infectious bursal disease (IBD) in the internal organs] were screened for the presence of IBD serotypes I, II and the variant strain by the agar gel immuno diffusion (AGID) test, employing standard antigens. Likewise the bursae, spleen and kidney specimen from 96 layer poultry with lesions suggestive of IBD were screened for the presence of IBD antigens employing standard antisera by the AGID test. The study revealed that antibodies to all three strains of IBD virus existed in 73.75% of sera from 400 broilers, while only serotype II antigen was detectable in tissues of the 96 layer birds that exhibited characteristic lesions of IBD. It was concluded that the serotype I vaccine did not protect the layer poultry against serotype II infection and the totally unvaccinated broiler poultry had suffered subclinical infection from all the three serotypes of IBD virus. PMID- 8403840 TI - An improved dot ELISA to detect fowl adenovirus type-1 antigen. AB - An improved dot immunobinding assay to detect fowl adenovirus type-1 is described. The method consists of spotting of antigen on nitrocellulose membrane sheet, blocking with either 5% acetic acid or with 5% defatted milk powder and fixation of either antigen or antigen-antibody complex, with either 50% methanol or 0.25% glutaraldehyde or 0.2% tannic acid. The results revealed that fixation of both antigen and antigen-antibody complex resulted in 4-fold increase in sensitivity when acetic acid was used as blocking agent. The use of two substrates simultaneously resulted in more colour intensity and clarity than using both substrates separately. PMID- 8403841 TI - Adventitious variability? The amino acid sequences of nonvertebrate globins. AB - 1. The more than 140 amino acid sequences of non-vertebrate hemoglobins (Hbs) and myoglobins (Mbs) that are known at present, can be divided into several distinct groups: (1) single-chain globins, containing one heme-binding domain; (2) truncated, single-chain, one-domain globins; (3) chimeric, one-domain globins; (4) chimeric, two-domain globins; and (5) chimeric multi-domain globins. 2. The crystal structures of eight nonvertebrate Hbs and Mbs are known, all of them monomeric, one-domain globin chains. Although these molecules represent plants, prokaryotes and several metazoan groups, and although the inter-subunit interactions in the dimeric and tetrameric molecules differ from the ones observed in vertebrate Hbs, the secondary structures of all seven one-domain globins retain the characteristic vertebrate "myoglobin fold". No crystal structures of globins representing the other four groups have been determined. 3. Furthermore, a number of the one-, two- and multi-domain globin chains participate in a broad variety of quaternary structures, ranging from homo- and heterodimers to highly complex, multisubunit aggregates with M(r) > 3000 kDa (S. N. Vinogradov, Comp. Biochem. Physiol. 82B, 1-15, 1985). 4. (1) The single-chain, single-domain globins are comparable in size to the vertebrate globins and exhibit the widest distribution. (A) Intracellular Hbs include: (i) the monomeric and polymeric Hbs of the polychaete Glycera; (ii) the tetrameric Hb of the echiuran Urechis; (iii) the dimeric Hbs of echinoderms such as Paracaudina and Caudina; and (iv) the dimeric and tetrameric Hbs of molluscs, the bivalves Scapharca, Anadara, Barbatia and Calyptogena. (B) Extracellular Hbs include: (i) the multiple monomeric and dimeric Hbs of the larva of the insect Chironomus; (ii) the Hbs of nematodes such as Trichostrongylus and Caenorhabditis; (iii) the globin chains forming tetramers and dodecamers and comprising approximately 2/3 of the giant (approximately 3600 kDa), hexagonal bilayer (HBL) Hbs of annelids, e.g. the oligochaete Lumbricus and the polychaete Tylorrhynchus and of the vestimentiferan Lamellibrachia; and (iv) the globin chains comprising the ca 400 kDa Hbs of Lamellibrachia and the pogonophoran Oligobrachia. (C) Cytoplasmic Hbs include: (i) the Mbs of molluscs, the gastropods Aplysia, Bursatella, Cerithedea, Nassa and Dolabella and the chiton Liolophura; (ii) the three Hb of the symbiont harboring bivalve Lucina; (iii) the dimeric Hb of the bacterium Vitreoscilla; and (iv) plant Hbs, including the Hbs of symbiont-containing legumes (Lgbs), the Hbs of symbiont-containing non-leguminous plants and the Hbs in the roots of symbiont free plants.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8403842 TI - Amino acid sequences of mammalian kazal-type proteinase inhibitors from salivary glands. AB - 1. The amino acid sequences of bikazins (the double-headed Kazal-type proteinase inhibitors from submandibular glands) isolated from the snow leopard (Unica unica), the European mink (Mustela lutreola), and the European pine marten (Martes martes) were determined. 2. N-terminal domains of bikazins are characterized by a cysteine residue spacing that differs from that of C-terminal domains of bikazins and other Kazal-type proteinase inhibitor domains. 3. N terminal sequences of bikazins seem to be specific for, and highly conserved within, each Carnivora family. PMID- 8403843 TI - Allozyme variation in a wild African elephant (Loxodonta africana) population from the Kruger National Park, South Africa. AB - 1. Blood, liver, heart, testis, skin, eye, muscle and kidney samples were obtained from elephants (Loxodonta africana) in the Kruger National Park during a culling programme in April 1992. 2. Gene products of 25 protein coding loci in L. africana were examined by horizontal starch-gel electrophoresis. 3. Eighteen protein coding loci (72%) displayed monomorphic gel banding patterns whereas only seven (28%) displayed polymorphic gel banding patterns. 4. Average heterozygosity values for adults, youngsters and the total population are respectively 0.058, 0.024 and 0.047. 5. Relative gene diversities within and between populations are 84% and 16% respectively. 6. Two population simulation programmes were utilized to predict the duration of the current variability present in this species, based on current genetic variation and gene transfer from one generation to the next. PMID- 8403844 TI - Relationship between structure and function of liver mitochondria from hibernating and active ground squirrels, Citellus undulatus. AB - 1. Electron microscopy of liver tissue preparations, obtained from hibernating ground squirrels, reveals mitochondria in a condensed state. 2. When kept on ice, mitochondria isolated from the livers of hibernating and active ground squirrels are in a shrunken state. 3. Incubation of mitochondria isolated from the livers of active ground squirrels in the presence of succinate, at 27 degrees C, results in mitochondrial swelling, while mitochondria from hibernating ground squirrels under the same conditions remain relatively shrunken. 4. The swollen mitochondria from active ground squirrels show high oxidative activity, while the shrunken mitochondria from hibernating animals show low oxidative activity. 5. Swelling of mitochondria from hibernating ground squirrels in a hypo-osmolar medium is accompanied by a significant increase in oxidative activity. 6. It is inferred that the shrinkage of hibernating ground squirrel mitochondria is one of the main causes of the inhibition of oxidative activity and other mitochondrial functions during hibernation. PMID- 8403845 TI - Comparative study on rat enteromucins. AB - 1. Mucus glycoproteins (mucins) were extracted from the duodenum, small intestine (proximal and distal parts) and colon. 2. Enteromucin subunits were larger than 2 x 10(6). 3. The glycopeptides of enteromucins were essentially the same in size of the subunit. 4. The carbohydrate portion of enteromucins consisted of N acetylgalactosamine, N-acetylglucosamine, galactose, fucose and sialic acid and the ratios of these sugars differed according to the region. 5. Average oligosaccharide length was about 8, 8, 8 and 14 sugars in the duodenum, proximal small intestine, distal small intestine and colon, respectively. 6. The proportion of threonine to serine in small intestinal mucins differed from duodenal and colonic mucins. Duodenal and colonic mucins were richer in threonine than small intestinal mucins. 7. Proximal and distal small intestinal mucins showed the same features. 8. A comparison of rat enteromucins to gastromucins indicated enteromucins to have large subunits and glycopeptides. The threonine and sialic acid content of enteromucins was higher and fucose content less compared to gastromucins. PMID- 8403846 TI - Prostaglandin-like activity, fatty acid and phospholipid composition of sika deer (Cervus nippon) antlers at different growth stages. AB - 1. The alteration of lipid composition has been shown to take place at different stages of antler growth. 2. The greatest amounts of phospholipids and polyunsaturated fatty acids have been found during the most intense soft antler growth period. 3. The bioregulators of lipid origin which are prostaglandins of A, B, E and F groups have been found at the same stage. PMID- 8403847 TI - Fibrinogenolytic proteases from the venoms of juvenile and adult northern Pacific rattlesnakes (Crotalus viridis oreganus). AB - 1. Venoms of Crotalus viridis oreganus show marked ontogenetic variation in protease activity. Adult venoms are approximately five-fold higher in protease (caseinolytic) activity. 2. Of seven potential protease inhibitors, only EDTA and 1,10-phenanthroline caused a significant decrease in protease activity. Responses of juvenile and adult venoms were essentially equivalent, and attempts at recovery of protease activity of EDTA-treated venoms by the addition of Ca2+ or Zn2+ were unsuccessful. 3. Gel filtration resolved two proteases from juvenile and subadult venoms with approximate M(r) of 100,000 and 78,000. Four proteases were resolved from adult venom, and M(r) estimates were 78,000, 61,000, 35,000 and 19,000. 4. Proteases from juvenile and adult venoms showed fibrinogenolytic activity, each producing some unique degradation products. 5. The occurrence of three "new" proteases in adult venom produced the ontogenetic increase in activity seen in the crude venoms. PMID- 8403848 TI - Carbonic anhydrase III content in various equine muscles. AB - 1. In this study, carbonic anhydrase III (CA-III) content in 18 equine muscles was determined by enzyme immunoassay. 2. It was found to differ in several muscles. 3. That in external intercostal muscle, rectus abdominis muscle and splenius muscle from four horses was very high. 4. Although the masseter muscle had only type I fibers, CA-III content was similar to that in mixed-fiber type muscles such as the biceps femoris muscle. 5. It thus appear that equine type I fibers can be further subgrouped. PMID- 8403849 TI - Comparison of transferrin sequences from different species. AB - 1. Amino acid sequences of transferrins from eight species, from human to tobacco hornworm, have been compared. Eighty-four amino acids (12%) are invariant, including three of the four ligands for the N-terminal Fe3+ ion. 2. The most highly conserved regions of both lobes of transferrin are the internal beta sheets of domains 1 and 2, and helices 5 and 7 which abut the Fe3+ binding site. Two small patches of conserved surface residues, which may be involved in receptor binding, have also been identified. 3. Phylogenies have been deduced from pairwise alignment of the sequences of the N- and C-terminal lobes independently. The phylogenies are consistent with the evolutionary tree derived from the fossil record, and with the observation that the gene duplication which created the N- and C-terminal lobes of transferrin occurred before the divergence of the mammalian and insect lines. 4. The phylogenies predict that the lactotransferrin family diverged some 200 Myr ago, after the separation of the lines leading to mammals and birds. In contrast, the phylogenies predict that melanotransferrin diverged before the separation of the mammalian and avian lines. 5. Sequence comparisons also suggest that the stoichiometry of the transferrin receptor:transferrin complex is 2:1. PMID- 8403850 TI - Comparative studies of the serum lipoproteins and lipids in some domestic, laboratory and wild animals. AB - 1. The concentration of lipoproteins and lipids in the sera of several species of healthy adult laboratory animals (guinea-pig, rabbit and rat), domestic animals (cattle, sheep, goat, horse and swine) and wild animals (deer, wild boar, mink and fox) have been investigated under physiological conditions. Special attention was paid to the lipoproteins which are precipitated with heparin-MnCl2 and to the ratio of lipids in separated alpha and beta lipoproteins. 2. Most of the studied animals had significantly lower concentrations of total lipoproteins, beta lipoproteins and cholesterol than those in man (P < 0.001). Only some wild animals (mink and fox) had concentrations of total lipoproteins, beta lipoproteins, phospholipids and cholesterol which were significantly higher than man's (P < 0.001). 3. The ratio of lipids in the separated lipoprotein fractions also differed between the examined animals. Thus, in the sheep, rabbit, guinea pig and wild boar most of the cholesterol (70-76%) was in beta lipoproteins. However, in the horse (60%), mink (60%), fox (65%), goat (70%) and cattle (73%) the high density lipoprotein fraction was the main carrier of cholesterol. 4. With the exception of the guinea-pig (42%) most of the serum phospholipids were found in the high density lipoprotein fraction. PMID- 8403851 TI - Plasma lipid transport in the horse (Equus caballus). AB - 1. Equine plasma contains lipoproteins corresponding to very low density (VLDL), low density (LDL) and high density lipoproteins (HDL). 2. HDL accounts for approximately 60% of plasma lipoprotein mass and consists of a single population of particles. 3. LDL is heterogeneous comprising three discrete subfractions. 4. Two proteins are found in the region of apolipoprotein (apo) B-100 in VLDL and LDL and a third similar to apoB-48 is in VLDL. 5. Lecithin:cholesterol acyl transferase is active in plasma and hepatic lipase and lipoprotein lipase are evident in post-heparin plasma. 6. There is no significant cholesteryl ester transfer protein activity. PMID- 8403852 TI - Cysteine proteinase from Bombyx eggs: role in programmed degradation of yolk proteins during embryogenesis. PMID- 8403853 TI - Serum chemistry profiles for Lechwe waterbucks (Kobus leche): variations with age and sex. AB - 1. Over an 8-year period, 19 biochemical parameters have been determined at various ages in the blood serum of 92 clinically healthy Lechwe waterbucks (Kobus leche), 33 males and 59 females. 2. Significant differences have been noted with age. In neonates, the lowest values of total proteins, glucose, creatinine, urea, AST, ALT and iron have been noted; the highest ones have been seen for cholesterol, alkaline phosphatase, calcium and phosphorus. 3. With regard to sex, raised values of glucose, urea, alkaline phosphatase and ALT, and lowered values of cholesterol, have been noted in juvenile females compared with males of the same age. 4. In adult females, higher levels of urea and cholesterol and lower levels of glucose, triglycerides and natrium have been recorded compared with males. 5. With sex and age, no significant changes have been found in the levels of GGT, magnesium, chlorides and copper. 6. Our findings are discussed with those abstracted from the literature for related species. PMID- 8403854 TI - Structural properties of Rapana thomasiana grosse hemocyanin: isolation, characterization and N-terminal amino acid sequence of two different dissociation products. AB - 1. The native Rapana thomasiana grosse hemocyanin is dissociated under mild conditions and fractionated into two dissociation products, RHSS1 and RHSS2, with an apparent molecular mass of approximately 250 and approximately 450 kDa, respectively. The two species are present in approximately equivalent amounts. SDS-PAGE analysis reveals that the latter component is a dimer of approximately 250 kDa polypeptide chains. 2. The amino acid compositions, as well as some spectroscopic properties of RHSS1, are very similar to those of RHSS2. After dissociation under mild conditions of the native hemocyanin both species preserve their capability of binding reversibly molecular oxygen. 3. RHSS1 and RHSS2 are sequenced directly from the amino-terminus for 15 and 20 steps, respectively. These parts of the two polypeptide chains are highly homologous but with microheterogeneity associated with some positions. They also exhibit high homology with the N-terminal region of subunits or functional domains of other gastropod Hcs. PMID- 8403855 TI - Glucose turnover in lean and obese rats of the SHR/N-cp and LA/N-cp strains. AB - 1. The relationship between hypertension, obesity, non-insulin-dependent diabetes mellitus and various parameters of glucose metabolism was studied. Lean and obese rats of the SHR/N-cp and LA/N-cp congenic strains were studied at four months of age. 2. Tritium and 14C-labeled glucoses were infused in one set of rats while tritiated water and 14C-labeled alanine were infused in a second group. 3. Glucose oxidation, turnover, conversion to glycogen, fatty acid synthesis, and alanine conversion to glucose were determined, as were blood pressure, pulse pressure and heart rate. 4. The presence of obesity influenced body weight, body fat, de novo fatty acid synthesis, organ weights, glucose mass, glucose oxidation, glucose synthesis, glucose carbon turnover and pulse pressure. 5. It had no effect on glycogen synthesis, tissue glycogen levels, blood glucose, glucose space, or blood pressure. 6. Strain differences were observed in final body weight, organ weights, blood pressure, pulse pressure, hepatic fatty acid synthesis, glucose mass, glucose space, glucose synthesis, liver glycogen levels and glucose conversion to muscle glycogen. 7. Strain-phenotype interaction effects were observed on glucose incorporation into hepatic glycogen, Cori cycle activity, hepatic de novo fatty acid synthesis, final body weight, fat pad weight, heart weight, and mean arterial pressure. 8. These results suggest that although obesity and hypertension are genetic traits in these rats, these traits are independent in their influence on the metabolism of glucose and the development of non-insulin-dependent diabetes mellitus. PMID- 8403856 TI - A view of medical informatics as an academic discipline. PMID- 8403857 TI - An interactive graphics system for real-time investigation and multivariate data portrayal for complex pedigree data systems. AB - GRIFFIN (graphics investigation of familial information), an interactive graphics system for exploratory investigation of data on individuals associated by familial relationships, was designed to provide genetic epidemiologists a flexible, rapidly responsive tool for viewing and guiding exploration of complex databases in the context of familiar pedigree structures. It graphically portrays both categorical and multivariate scalar data on individuals in those structures. The display can be inverted to show all ancestors and descendants of any individual the user designates. It provides cues to censored information when bushy pedigrees cannot be fully displayed without sacrificing legibility. These guide users on where to next invert the system. Investigators may translate/zoom the display, vary the mode of representing data, point to individuals to obtain displays of alphameric information about them, etc. Developed in Fortran using IBM's GDDM graphics subroutines for an IBM 3090 mainframe, GRIFFIN's design anticipates porting to smaller systems. PMID- 8403858 TI - Acyclic directed graphs for automatic image analysis of lung parenchymal geometry. AB - We have developed a simple method for encoding pulmonary alveolar geometry suitable for morphometric analysis and modeling. The approach involves the use of 8-bit video microscopic images of lung which are thresholded to produce binary images. Each alveolar wall was thinned to its midline to produce a skeletonized image. We present an algorithm for converting such an image to an acyclic directed graph. Such a representation is quite suitable for mathematical treatment, lending itself to problems involving simulation or modeling. We present as an example application, the measurement of dihedral angles in rat lung using conventional histological sections. PMID- 8403859 TI - A time-series approach to predict outcome from pediatric intensive care. AB - Daily assessment of the physiologic status of intensive care unit (ICU) patients by a validated score is used to predict their discharge as either alive or dead. Daily scores were decorrelated by time-series analysis techniques to establish a predictor of the next day's score. Data from 3299 patients (290 deaths) in nine different pediatric ICUs were used to derive the predictor model. Using the highest predicted score observed in any survivor from this data base as a threshold value, this predictor correctly identified 21.9% (sensitivity) of the nonsurvivors with no errors in predicting a fatal outcome (100% specificity). This performance is significantly (P < 0.001) better than that with a static predictor based on the first- or second-day scores only (6.6 or 7.4% sensitivity, respectively), or a known empirical dynamic model using the scores of the 2 most recent days (10.3% sensitivity, P < 0.002). PMID- 8403860 TI - A computer program for estimating individualized probabilities of breast cancer. AB - A computer program is presented for estimating the absolute risk of developing breast cancer over a specified age interval for women with combinations of five risk factors, namely age at menarche, age at first live birth, family history of breast cancer, number of previous breast biopsies, and presence of atypical hyperplasia in biopsy specimens. Statistical methods have been developed and applied to data from the Breast Cancer Detection and Demonstration Project to obtain (i) point estimates of absolute risk by combining relative risk estimates from case-control data and estimated composite incidence rates from cohort data, and (ii) confidence intervals by using implicit delta-method arguments. The program is interactive and easy to use and is therefore well suited to assist in medical counseling. For instance, women with high estimated risk might be advised to undergo a program of frequent surveillance with mammography. An executable version for IBM-compatible PCs is available from the author upon submission of a PC diskette formatted with MS-DOS. PMID- 8403861 TI - Comparative study of human expertise and an expert system: application to the diagnosis of child's meningitis. AB - We present a general method of statistical evaluation for expert systems, applied to a system for the diagnosis of child's meningitis. Fifty difficult clinical cases of child's meningitis were submitted to the system, to seven senior specialists and to seven young physicians. Multidimensional analysis of the diagnosis of the infection category reveals that the two groups of physicians separate naturally and that the system is located among the group of experts. The study of the agreement of the microbiological diagnosis and therapeutic advice shows that the advice of the two groups of physicians is significantly different and that the advice of the system is significantly closer to that of the experts. This result is confirmed by the study of therapeutic errors. This type of study allows one to classify the performance of the system among physicians having different levels of expertise without referring to an objective solution. PMID- 8403862 TI - Kinetic analysis of epithelial cell migration in the colon on a massively parallel processor (CM-2). AB - A cell kinetic model of proliferating tissues and its implementation on the massively parallel computer, Connection Machine Model 2 (CM-2), is presented. The model is applicable to all proliferating tissues of the organism and is illustrated on the crypt-villus unit (CU) of the gastrointestinal mucosa. The crypt unit consists of two compartments, a progenitor (P) in which cells proliferate, and a proliferative quiescent (Q) compartment, in which cells are incapable of synthesizing DNA and therefore do not divide. Cells are formed in P, enter Q, traverse it, and die at its outer boundary. The healthy CU maintains a steady state; cell proliferation is controlled by cell elimination. The crypt cylinder is mapped onto a two-dimensional CM-2 matrix such that each processor represents a cell in the unit. The interprocessor links serve as proliferation controls. Using cell counts, the frequency of P cells in each cell location along the crypt column was estimated and various columnar kinetic parameters were calculated. The kinetic analysis shows that cell migration occurs along the crypt column in the direction of the colonic lumen at a velocity averaging 0.244 cell locations per hour. In order to compare model results with experimental kinetic studies, the model was used to stimulate a labeling experiment. The results are in agreement with published kinetic experiments that were done on a normal CU. The model allows three-dimensional visualization of the spatial and temporal events within the crypt. PMID- 8403863 TI - A simple program in BASIC to compare regression lines by analysis of covariance. AB - Most of the biological processes can be described by linear relationships. The comparison of regression lines is therefore very common in biological research. This paper presents a program written in BASIC that compares the slopes, intercepts and lines of regressions, using analysis of covariance. PMID- 8403864 TI - A computer program for regression analysis of repeated measures using generalized estimating equations. AB - RMGEE is an easy-to-use FORTRAN program for the analysis of repeated binary, count, and normally-distributed response variables using the generalized estimating equations approach of Liang and Zeger [1]. The program can be used when measurements are obtained at multiple time points from each subject or experimental unit, and also when the basic sampling unit is a group or cluster of subjects, and the response variable of interest is obtained from each subject within the cluster. It is not necessary for the number of repeated measurements to be the same for every experimental unit and missing data are easily accommodated. Both time-independent (cluster-specific) and time-dependent (occasion- or subject-specific) covariates are permitted. The program can be run on microcomputers, workstations, and mainframe computers. Three examples illustrating the usage and features of RMGEE are provided. PMID- 8403865 TI - An inexpensive computer-aided morphometric device for the analysis of dissociated brain cell cultures. AB - Morphometric analysis of brain cells growing in dissociated cell culture is a promising approach in neuroscience research. Here we present new software written in Turbo PASCAL which has the advantage of requiring only a minimum in equipment accessible in most neuroscience labs, i.e., a microscope with a camera lucida attachment and a personal computer. The camera lucida is used to superimpose the pictures from the microscope and the computer screen. With the aid of a mouse, cells are drawn to the computer screen. The program counts pixels and calculates morphometric data including the area occupied by the cell soma as well as number, length and arborization of the processes. The program may be obtained from the authors upon request. PMID- 8403866 TI - Computer-assisted individual estimation of radioiodine thyroid uptake in Grave's disease. AB - A computer-assisted Bayesian individual estimation of radioiodine thyroid uptake kinetics for patients suffering from Grave's disease is proposed. The program provides a fast computation of the activity to be administered to a given patient to achieve a target thyroid absorbed dose. This determination relies upon the patient biological covariates and upon a small number of measurements performed during a preliminar kinetic study of radioiodine thyroid uptake. Our results indicate that a two-sample Bayesian approach is reliable when external thyroid counts are performed at 2 h and 168 h after a test dose and has advantages over conventional kinetic experiments in terms of patient acceptability. This method is implemented on widespread computers and interfaced with a patient database. An interactive user interface with in-line data checking is provided. The program could be also a tool to better study the relationship between the absorbed dose and the clinical effect. PMID- 8403867 TI - ADAMS: Aggregate Data Management System for epidemiologists and health-care managers. AB - An effective health-care policy is supported by the availability of data in the form of statistical tables for epidemiologists and health-care managers. Creation, analysis and exploitation of these data strongly support the monitoring of trends in mortality and morbidity phenomena and the evaluation of offered health services. This might imply the transformation of statistical tables to more suitable formats. A successful management, manipulation and querying of a statistical table is a complex activity requiring a considerable knowledge of statistical problems. Computerised support can be precious in this task and this led to the development of several data management systems. In this paper we sketch out the features of ADAMS (Aggregate Data Management System), a system conceived to allow an easy interaction with statistical tables, reshaping and browsing of their descriptive part. ADAMS aims at simplifying the problem of extracting information from a statistical database and performing statistical table manipulation. The following functionalities are provided: definition of a statistical database at descriptive level; storage of data in the database; manipulation of data according to the summarisation and reclassification operators; browsing between data and their descriptive part. It also supports different manipulation styles according to different user profiles. PMID- 8403868 TI - DECIDE: a software for computer-assisted evaluation of diagnostic test performance. AB - The evaluation of the performance of clinical tests is a complex problem involving different steps and many statistical tools, not always structured in an organic and rational system. This paper presents a software which provides an organic system of statistical tools helping evaluation of clinical test performance. The program allows (a) the building and the organization of a working database, (b) the selection of the minimal set of tests with the maximum information content, (c) the search of the model best fitting the distribution of the test values, (d) the selection of optimal diagnostic cut-off value of the test for every positive/negative situation, (e) the evaluation of performance of the combinations of correlated and uncorrelated tests. The uncertainty associated with all the variables involved is evaluated. The program works in a MS-DOS environment with EGA or higher performing graphic card. PMID- 8403869 TI - Airport X-rays and floppy disks: no cause for concern. AB - A controlled study was done to test the possible effects of X-rays on the integrity of data stored on common sizes of floppy disks. Disks were exposed to doses of X-rays up to seven times that to be expected during airport examination of baggage. The readability of nearly 14 megabytes of data was unaltered by X irradiation, indicating that floppy disks need not be given special handling during X-ray inspection of baggage. PMID- 8403870 TI - A microcomputer based lung sounds analysis. AB - The use of a microcomputer in lung sound-analysis is described. The system was used experimentally in order to evaluate automated auscultation as a mean for improving the sensitivity of pulmonary health mass screening. The sound signals from four custom-made piezoelectric transducers, affixed at specific locations on the chest wall, and the breathing flow signal produced by a pneumotachograph were amplified, filtered and digitized simultaneously at 4000 Hz per channel for 512 ms. The acoustic data were transformed to the frequency domain to enable the calculation of the power spectra. Those were averaged over successive runs and displayed as log power vs. frequency. The operator could assess the convergence of the spectral pattern using the on-line graphics and calculated parameters, and store the data once the noise level had reached a preset level. This procedure was repeated during expiration, inspiration and on breath arrest. The results of the off-line analysis of the lung sounds, combined with pulmonary function tests and a questionnaire, were used to identify lung pathology. PMID- 8403871 TI - Special computer graphical method for detection of myocardial infarction. PMID- 8403872 TI - [Nursing research in the United States]. PMID- 8403873 TI - [Case study reports, No. 22 (Society for the Study of Scientific Nursing)]. PMID- 8403875 TI - [Asking about 'nursing and medicine']. PMID- 8403874 TI - [An introduction to Nightingale's nursing]. PMID- 8403876 TI - [A visit to the Nightingale Museum]. PMID- 8403877 TI - [A trial in educational development of the general clinical nursing theory]. PMID- 8403878 TI - [Searching for logic of children's recognition]. PMID- 8403880 TI - [What is psychiatry?]. PMID- 8403879 TI - [Beginning course of English medical terminology based on analytical method]. PMID- 8403881 TI - [The world of Hildegard von Bingen]. PMID- 8403882 TI - Atypical seminoma--histologic features, immunohistochemical characterization, and correlation with survival. AB - Classical seminoma and embryonal carcinoma are two points in the spectrum of histologic differentiation in testicular germ cell tumors. The validity of an intermediate category, ie, atypical seminoma (AS) is questionable. Histopathologic and clinical data on 42 patients treated for primary testicular germ cell tumor from 1975 to 1985 were reviewed. Twenty-seven cases were identified as classical seminoma and nine were embryonal carcinoma. The remaining six cases were somewhat problematic to classify, combining the growth pattern of seminoma with cytologic features of embryonal carcinoma. Immunocytochemically, four of these tumors suggested some progression towards the embryonal carcinoma phenotype on the basis of cytokeratin expression. Survival for classical seminoma, AS, and embryonal carcinoma were 90%, 80%, and 63% respectively (mean follow-up, 8.6 years). Although the survival differences were not statistically significant, when considered with morphologic and selected immunocytochemical data, they tend to support the concept of an AS as an intermediate lesion between classical seminoma and embryonal carcinoma. PMID- 8403883 TI - Acute acalculous cholecystitis associated with common hepatic duct obstruction: a variant of Mirizzi's syndrome. AB - A variation of Mirizzi's syndrome is described in which common hepatic duct obstruction occurs due to extrinsic compression by an acutely inflamed acalculous gallbladder abutting the common hepatic duct. This case suggests that extrinsic compression and inflammation in the absence of stones can result in complete obstruction of extrahepatic biliary flow. PMID- 8403884 TI - Henry Gray of legendary textbook fame. PMID- 8403885 TI - Dealing with disciplinary hearings before Connecticut's Department of Health Services. PMID- 8403887 TI - Dedicated volunteers provide medical aid for Stamford's homeless. PMID- 8403886 TI - Physician practice acquisitions and health care reform: meeting the Office of Inspector General's fraud and abuse concerns. PMID- 8403888 TI - Toxicity of sodium nitroprusside. PMID- 8403889 TI - The Brody plan. PMID- 8403890 TI - 1993 AMA principles of medical ethics. PMID- 8403891 TI - The five five's. PMID- 8403892 TI - Detection of a type IX collagen-related mRNA in an invertebrate, the marine annelid Nereis virens. AB - Fibrous and non-fibrous collagens have been described in both vertebrate and invertebrate animals. However, there has been limited characterization of non fibrous collagens and their corresponding genes in invertebrate animals. In the present study we have used as a probe an avian cDNA clone which encompasses the COL3, NC3 and part of the COL2 domain of the collagen alpha 3(IX) subunit. This probe hybridized to mRNA obtained from the cuticle and body of the marine annelid, Nereis virens. Northern blot hybridization exhibited an mRNA of ca. 7.5 8 kilobases which in situ hybridization shows to be most abundant over cuticle associated cells. Dot-blot hybridization, comparing cuticle mRNA and body mRNA, indicates that this collagen mRNA is five times more abundant in the cuticle. The composite data suggest evolutionary conservation, in both vertebrate and invertebrate animals, of a non-fibrillar collagen. PMID- 8403893 TI - Biochemical changes in the collagen of human osteoporotic bone matrix. AB - Although it is known that collagen imparts mechanical strength to bone no detailed biochemical analysis has been made of osteoporotic bone collagen. We report for the first time significant changes in the properties of the collagen. Analysis of collagen types revealed little change in the proportion of Type III collagen, but in some cases there was a significant loss of the Type VI. However, the major differences were observed in the post-translational modifications, namely, in the stabilizing cross-links and the hydroxylation of the collagen. These changes indicated a higher turnover in the head region compared to the neck region of the femoral head and are consistent with the susceptibility of the neck region to fracture. Clearly, the collagen is altered in osteoporosis and these changes may play a role in the pathogenesis of the disease. PMID- 8403894 TI - Changes in the ratio of non-calcified collagen to calcified collagen in human vertebrae with advancing age. AB - Bone loss associated with aging is associated primarily with a decline in bone formation. To try and further understand the nature of this process we have used a biochemical approach which relies on the fact that osteoid is susceptible to enzymatic degradation whereas calcified collagen is protected by the mineral phase against proteolytic digestion. Our findings show a statistically significant inverse relationship between osteoid and age (r = 0.70 female, r = 0.47 male). A closer relationship was observed when age was related to the ratio of osteoid to bone (r = 0.73 female, r = 0.56 male). In both cases, the observed linear decline begins at an early age and becomes marked with advancing age. Histologic observations illustrate these findings showing decreased osteoid and osteoblasts in the older vertebral specimens compared to the younger ones. Even though the mechanism for osteoid calcification seems to remain unimpaired, the decline of a calcifiable matrix in the presence of normal bone turnover could lead to bone loss. PMID- 8403895 TI - Tendon collagens: extracellular matrix composition in shear stress and tensile components of flexor tendons. AB - Outer synovial tissues were separated from the remainder of avian flexor tendon and the collagens characterized biochemically and compared with those of the internal portion of tendon and sheath. The collagen content of tendon synovium was 23%, whereas that of tendon and sheath were 78% and 73%, respectively, based on dry weight. Four genetic types of collagen were found in the pepsin solubilized matrices: in the synovium, types I (78%) and III (19%) predominated; types V and possibly VI were present as minor collagens. Purified synovial type V collagen was a heterotrimer, with chain composition [alpha 1(V)]2 alpha 2(V). In contrast, the internal portion of tendon and sheath were comprised of only type I collagen. There was a large amount (41%) of ethanol extractable, noncollagenous material present in synovium, a part of which was proteoglycans. In addition, collagen cross-links of these tissues were quantified: the internal tendon had an abundant concentration of pyridinoline; synovium exhibited high amounts of labile, reducible cross-links, particularly dihydroxylysinonorleucine. In the case of sheath, lysine aldehyde-derived cross-links appeared to be predominant. These results indicate that each tissue has its own collagen type distribution as well as cross-linking pattern reflecting their maturational and functional differences. PMID- 8403896 TI - Is the antioxidant, anti-inflammatory putative new vitamin, PQQ, involved with nitric oxide in bone metabolism? AB - Our laboratory recently isolated free PQQ (2,7,9-tricarboxy-pyrroloquinoline quinone, methoxatin), a bacterial redox cofactor, from red cells, neutrophils, serum and milk and found free PQQ in CSF, synovial fluid and bile. The metabolism and functions of PQQ and ascorbate may be coupled. Physiologically, free PQQ catalyzes dioxygen-superoxide interconversion, and participates in both superoxide generation (respiratory burst) and scavenging (cell protection). Using a labeled aromatic o-diamine, superoxide formation by activated neutrophils was inhibited and the labeled phenazine adduct of PQQ could be isolated from the inhibited cells (Karnovsky et al., 1992). PQQ may convert xanthine oxidase to xanthine dehydrogenase (XD) and could be the physiological coenzyme of XD. PQQ plus copper, form a potent amine-oxidizing system. Shah et al., 1992 found that PQQ-Cu2+ catalyzes the oxidation of epsilon-amino groups in collagen and elastin. Rucker's lab (Smidt et al., 1991) has found that PQQ may be a vitamin for mouse pups. Watanabe et al., 1988 and Nishigori et al., 1989, showed that injected PQQ protects animals against oxidative stress injury. PQQ's in vivo antioxidant action, spares reduced glutathione. PQQ, as an actively transported organic anion, concentrates in cells. In other experiments (Aizenman et al., 1992), PQQ protected neurons against the neurotoxin action of the glutamate-receptor against NMDA. We shall consider possible roles for PQQ in the biosynthesis of nitric oxide (NO, endothelium-derived relaxing factor, EDRF) from L-arginine and in NO removal by superoxide. NO has now been linked to the inhibition of osteoclastic bone resorption. PMID- 8403897 TI - Immuno-identification of two non-amelogenin proteins of developing bovine enamel isolated by affinity chromatography. Further proof that tooth "enamelins" are mainly serum proteins. AB - Affinity chromatography of "Enamelin Extracts" of developing bovine molar enamel on CNBr activated Sepharose 4B to which polyclonal antibodies of whole bovine serum and fetuin were cross-linked, revealed that at most, only 1-2% of the proteins in the extracts were not bound to the columns. The approximately 98% or more of the proteins in such extracts were bound to the resin and were eluted in the position of the serum proteins and fetuin. The small amount of protein which was not bound to the affinity column and which was eluted very early, was subjected to SDS-PAGE and immunostained with polyclonal antibodies to two non amelogenin proteins isolated and identified previously (approximately 26kDa and 22kDa). The antibody to the approximately 50kDa protein immunostained strongly with only one protein band of approximately 26kDa. The antibody to the approximately 22kDa band reacted strongly with one protein band of approximately 22kDa, weakly with an approximately 100kDa and very weakly with several lower molecular weight bands, suggesting either aggregation and degredation of the 22kDa component, or more likely that the approximately 22kDa and lower molecular weight proteins are all derived from the approximately 100kDa component. There was no immuno-crossreactivity of the 22kDa and 26kDa antibodies and none of the components eluted in the early first fraction reacted with polyclonal antibodies to amelogenins or to amelogenin peptides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403898 TI - The post-translational chemistry and molecular packing of mineralizing tendon collagens. AB - Using cross-linking stereochemistry as indicators, the molecular environment of two collagens in the turkey leg Achilles tendon were compared. The tendon from one year old turkeys was dissected into nonmineralized, fully mineralized and transitionally mineralized portions. Amino acid composition and cyanogen bromide peptide mapping of these portions indicated that the collagens were essentially type I throughout. The fully mineralized compartment had a lysine hydroxylation level similar to turkey or mammalian bone collagen. The non- and transitionally mineralized collagens had a significantly higher lysine hydroxylation, typical of tendon or ligament. However, unlike mammalian tendon, the collagen cross-links were essentially derived from the carboxy-terminal ends of the molecules. The predominant cross-link in this portion was pyridinoline having a high content of 0.95 +/- 0.09 res/mole of collagen. The cross-links in the fully mineralized collagen were also essentially derived from carboxy-terminal aldehyde. However, here significant amounts of the lysyl analog of pyridinoline and lysine-involved bifunctional cross-links were present. The molecular loci of pyridinoline in nonmineralized collagen and the lysyl analog of pyridinoline in mineralized collagen were found to be identical. The total trifunctional cross-link level in the mineralized collagen, 0.55 +/- 0.05 res/mole of collagen, was virtually identical to that observed in old mammalian bone and dentin, and in long term in vitro incubation studies of predentin. We have tentatively concluded that the post-translational chemistry and molecular environments are different in these two turkey tendon fibrils. However, a relative paucity of amino-terminal based cross-links is a feature they have in common. The possible involvement of the amino-terminal telopeptides in collagen mineralization is discussed. PMID- 8403899 TI - Amino-terminal location of pyridinoline in dentin collagen. AB - Cross-linking is believed to be one of the major factors that characterize the calcifiability of dentin and bone collagens. Dehydro-dihydroxylysinonorleucine and pyridinoline which constitute the principal cross-links of dentin collagen have so far been located only in the carboxy terminal telopeptide of the molecules [alpha 1(I)-chain 87 x alpha 1(I)-chain 16C]. This situation suggested that the amino terminal telopeptide portion might be "open" without intermolecular cross-linking in hard tissue collagen fibrils. However, the present study provided evidence that pyridinoline is also located in amino terminal telopeptides (alpha 1-chain 9N or alpha 2-chain 5N) and alpha 1-chain 930. Bovine dentin collagen was digested with trypsin followed by heating at 60 degrees C before and after the digestion. This method gave complete trypsin peptides of dentin collagen. Fluorescent pyridinoline peptides with a smaller molecular size were isolated by Sephadex G-50 superfine, DEAE-cellulose and reverse-phase HPLC. Automatic Edman analysis of several isolated peptides revealed the five-residue sequence, Gly-Ile-X-Gly-His-Arg, the only assignment of which was alpha 1-chain 928-933. The above evidence together with the amino acid compositions of the peptides led to the conclusion that pyridinoline is located not only in the carboxy-terminal but also in the amino-terminal telopeptide in dentin collagen. PMID- 8403900 TI - Another look at the Dalkon Shield: meta-analysis underscores its problems. AB - Numerous non-comparative clinical trials of the Dalkon Shield appear to provide ample evidence that the Dalkon Shield was an effective IUD; they seem to yield little evidence that it was a dangerous device, nor markedly different from its contemporaries. Equating the performance of the Dalkon Shield IUD to that of the Lippes Loop or of Copper IUDs is erroneous, however, with respect to rates of pregnancy, expulsion, pelvic infection, septic abortion, death with the device in situ, and tubal infertility. Randomized studies show the Dalkon Shield had approximately double the pregnancy rates of the Lippes Loop D or Copper IUDs (P < .05) and a significantly lower expulsion rate. These differences coupled together with the distinguishing multifilament tail of the Dalkon Shield underlay the higher rates of pregnancy complications, including septic abortions with the device in situ and deaths, associated with the Shield. A five-fold increased risk of hospitalized pelvic infection among Dalkon Shield users found in the Women's Health Study resulted not from ascertainment bias, but was related to the fact that Dalkon Shield users had more severe hospitalized PID than did other hospitalized women with PID and IUD use. Following cessation of distribution and of use of the Dalkon Shield, and following the FDA's recommendation to remove IUDs in case of pregnancy, there have been no deaths reported among pregnant American women with an IUD in situ in a 15-year period. Neither the IUDs of today nor those in use during 1970-1974 are equitable to the Dalkon Shield [corrected]. PMID- 8403901 TI - Factors associated with copper T IUD removal for bleeding/pain: a multivariate analysis. AB - To examine the factors associated with copper T IUD removal due to bleeding/pain, a nested case-control analysis using data from an international multicenter randomized clinical trial of the copper T-380A IUD was performed. One-hundred forty-three cases with removal due to bleeding/pain within one year postinsertion and 2,023 controls with the IUD in place at last follow-up visit are included. Proportional hazards regression analysis indicated that uterine length greater than 70 mm and better IUD provider's skill were associated with a decreased risk of removal due to bleeding/pain. Amenorrhea and breastfeeding at insertion have short-term beneficial effects but less long-term effects. These findings may help family planning providers in counseling and practice. PMID- 8403902 TI - Performance of the copper T 200 in parous adolescents: are copper IUDs suitable for these women? AB - The clinical performance of a cohort of 995 parous adolescents, first-time users of the T-Cu 200B was studied and compared with a cohort of paired controls 10 years older, of the same parity. Life-table analysis showed that pregnancy, expulsion rates, as well as removals for bleeding and/or pain were higher in adolescents but within ranges already reported in the literature on this device. Removals for infection were few and not significantly different. Although the clinical performance of the T-Cu 200B in adolescents is not as good as in older women, the performance is similar or better than that reported for other reversible methods in this age group. Our conclusion is that IUDs should not be contraindicated for parous adolescents as long as adequate counseling, screening of risk factors, skillful insertion and follow-up are provided. PMID- 8403903 TI - Effectiveness of the diaphragm, used continuously, without spermicide. AB - The experience of 670 diaphragm users from three Brazilian clinics was retrospectively reviewed. Most of the subjects (441) followed the traditional method (TM) of using the diaphragm, only at the time of sexual intercourse and with spermicide. One third (215) used the diaphragm continuously (CU), without spermicide, removing it only at the time of their daily shower to wash it, with immediate re-insertion. The total pregnancy rate was 7.0 per 100 women-years for the whole group. Patient and total failure rate were significantly lower in the CU (0.6 and 2.8), in comparison with the TM sub-group (6.5 and 9.8). The discontinuation rate for urinary infection or other medical reasons was not greater in the CU group. Logistic regression analysis, including age, parity, number of abortions and of living children, years of schooling, whether living with a partner, presence of mild cystocele or retroverted uterus, and form of use of diaphragm (TM or CU), showed that only the TM of diaphragm use was positively associated to patients' and total failure rate. PMID- 8403904 TI - The effects of a long-acting progestin on the hypothalamic-pituitary-ovarian axis in women with normal menstrual cycles. AB - This study was performed to determine how a long-acting, slow-release preparation of norethindrone (NET) affects the hypothalamic-pituitary-ovarian axis of normal ovulatory women. Ten women were studied during the luteal phase of their menstrual cycle, and again at six and twelve weeks following intramuscular administration of 100 mg NET microencapsulated in poly-D,L-lactide-co-glycolide. Serial LH samples, serum E, P, and NET were followed by a GnRH stimulation test. Compared to luteal phase values, six and twelve weeks of treatment with NET inhibited serum E2 and P while mean serum LH remained unchanged and mean serum FSH increased significantly (p < 0.05). LH pulse frequency after NET treatment was twice the rate (p < 0.01) as that of the luteal phase, whereas LH pulse amplitude was decreased significantly (p < 0.05). Finally, although there was no significant change in pituitary LH secretion in response to GnRH, NET treatment augmented FSH responsiveness to GnRH at the times studied. Preserved pituitary responsiveness to GnRH in NET-treated patients suggests that inhibited ovarian function results in an increase in GnRH pulse frequency but not GnRH pulse amplitude. Since the progestational milieu is maintained in these patients by NET treatment, the decrease in serum E2 may be responsible for the increase in GnRH pulse frequency. The presence of a critical level of E2 may be necessary for progestins to affect the hypothalamic GnRH pulse generator. PMID- 8403905 TI - Hormonal and clinic evaluation of patients with moderate body hair growth. AB - Plasma prolactin (PRL), gonadotropins (FSH, LH), estradiol-17 beta (E2), progesterone (P), total testosterone (T), sex steroid binding protein (SBP), T/SBP index, cortisol (F), 17-OH-progesterone (17OH-P), dehydroepiandrosterone sulphate (DHEA-S) and androstenedione (A), were measured in 50 fertile non-obese women presenting with moderate body hair growth and in 30 matched controls. DHEA S and PRL were significantly higher (P < 0.002, P < 0.001, respectively) and SBP was lower (P < 0.001) in patients than in controls. Regression analyses showed that PRL levels were independent of the other parameters, while a negative correlation was found between DHEA-S and SBP values. Since the decision to treat a woman with mild body hair growth is usually a clinical one, PRL behaviour has to be taken into account before deciding the type of treatment. Clinical improvement was observed in subjects treated with ethynylestradiol plus desogestrel or plus cyproterone acetate, so as to produce an increase in SBP rather than a decrease in DHEA-S. PMID- 8403906 TI - Treatment with a progesterone antagonist ZK 98.299 delays endometrial development without blocking ovulation in bonnet monkeys. AB - The effects of an antiprogestin ZK 98.299 (onapristone) on serum levels of estradiol and progesterone, and on the endometrial morphology were studied in adult bonnet monkeys. Twelve animals having menstrual cycles of normal duration (24 to 30 days) were randomly distributed into 4 equal groups. The animals in Group 1 were treated (s.c.) with the vehicle (benzyl benzoate: castor oil, 1:10), and in Groups 2, 3 and 4 with 5 mg, 10 mg, or 20 mg ZK 98.299 once-a-week, respectively. Treatment was initiated on day 1 of the menstrual cycle and each animal in Groups 1, 2 and 3 was treated for two consecutive cycles. Since the treatment cycle length of animals in Group 4 was considerably prolonged, they were treated for one menstrual cycle only. Endometrial biopsy was taken around day 20 of the second treatment cycle of first three groups and around day 50 of the 4th group of animals. Treatment with vehicle or 5 mg ZK 98.299 had no significant effect on the menstrual cycle length. Treatment with 10 mg dose had no effect in two animals and prolonged the cycle length in one, whereas, further increase in the dose to 20 mg prolonged the cycle length in all the animals. The duration of menses was generally reduced. Treatment with vehicle or different doses of ZK 98.299 had no effect on ovulation. In animals treated with 5 or 10 mg dose, the pattern of mid cycle rise in serum estradiol levels and progesterone levels during the luteal phase of both treatment cycles were comparable to those of vehicle-treated animals and were suggestive of normal ovulatory cycles. On the other hand, in animals treated with the higher dose (20 mg/week), progesterone levels during the luteal phase were significantly reduced and were indicative of luteal insufficiency. The hormonal data during the treatment period of this group of animals was suggestive of two distinct ovarian cycles indicating that the menstrual bleeding during the treatment period was probably very scanty. Treatment with ZK 98.299 impaired the endometrial development in a dose-dependent manner. In vehicle-treated animals, the endometrium had large and tortous glands with secretions. Treatment with ZK 98.299 caused atrophic changes in the glands as well as in the stroma. The height of the epithelial cells was markedly decreased and they became small and inactive. This study, therefore, suggests that treatment with low doses of antiprogestin ZK 98.299 at weekly intervals does not block folliculogenesis or ovulation, but has an inhibitory effect on the endometrium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8403907 TI - Arterial sodium concentration in rats with hypertension induced by mestranol. AB - Rats were made hypertensive by the oral ingestion of mestranol for 6 months. Carotid arteries from these hypertensive rats and from control rats were incubated in an extract of plasma from these rats, which contained 24Na and 14C sucrose (an extracellular fluid marker). After sufficient time for equilibration, the vessels were removed and counted for 24Na; an aliquot of the incubation fluid was also counted for 24Na. At least 7 days later, after the 24Na had decayed away, the samples were counted for 14C-sucrose. By knowing the specific activity of the 24Na and the ratio of 24Na/14C in the bathing fluid, the amount of intracellular Na could be calculated for each carotid artery. The total protein content of each vessel was determined, and the results were expressed as nEq of Na per mg of vessel. The Na content of carotids from mestranol treated rats averaged 135 +/- 11 (SEM) while the carotids from the control rats averaged 125 +/- 8 nEq/mg protein. Because these values were not significantly different, this study provided no evidence that increases in arterial Na content contributed to the hypertension associated with the ingestion of mestranol in this rat model. PMID- 8403908 TI - A comparative study of two low-dose combined oral contraceptives: results from a multicenter trial. AB - A comparative multicenter clinical trial of two low-dose combined oral contraceptives (OCs) was conducted in Malaysia, Egypt, Thailand, and Mexico. Efficacy, safety and acceptability were investigated in women taking either a norgestrel-based (NG) OC or a norethindrone acetate-based (NA) OC. This paper includes analysis of 892 women, all of whom were at least 42 days but within 26 weeks postpartum and randomly allocated to one of the above OCs. Follow-up visits were scheduled at 1, 4, 8 and 12 months after admission. Baseline sociodemographic characteristics were similar for both groups, as well as compliance. There were nine unintended pregnancies reported; eight of these occurring in the NA group. Adverse experiences were minor with headaches and dizziness being the most common complaints; frequency of reports was similar in both groups. The group taking the NG-based OC had significantly (p < .05) fewer menstrual-related complaints. Discontinuations due to menstrual problems were significantly more common among NA users (primarily amenorrhea). Discontinuations in the NG group were primarily for other personal reasons, e.g. unable to return to the clinic. There was also a significant difference between the two groups for the 11-month gross cumulative life table discontinuation rates due to menstrual problems (p < .01); the NA group had the higher rate. PMID- 8403909 TI - Phase III clinical trial with Norplant II (two covered rods): report on five years of use. AB - The subdermal implant NORPLANT II contraceptive was studied for its safety, efficacy and acceptability over a period of 5 years of use in a phase III multicentre clinical trial. A total of 1,466 women were observed for 52,849 women months of use. Only four pregnancies were reported during the study period, giving a method failure rate of 0.8 per 100 users at 5 years of use. The continuation rates were 61.4, 49.0 and 42.1 per 100 users at 3, 4 and 5 years of use, respectively. The majority of the discontinuations were due to bleeding irregularities which accounted for 22.2, 26.3 and 28.5 per 100 users at 3, 4 and 5 years of use, respectively. The next common reason was planning pregnancy which was observed mainly in women having one child. The discontinuations due to infection, expulsion or displacement of device were very low (0.2-0.3 per 100 users). Due to vigorous efforts made by the centers to follow the subjects, the lost-to-follow-up rate was very low (1.6% at 5 yr). PMID- 8403910 TI - Pharmacokinetic study of RU 486 and its metabolites after oral administration of single doses to pregnant and non-pregnant women. AB - RU 486 and three of its metabolites (RU 42633-monodemethyl, RU 42848-didemethyl, and RU 42698-hydroxymetabolite) were determined by HPLC in plasma from nine non pregnant and 36 pregnant women. Each non-pregnant subject took an oral dose of RU 486 (25, 100, 400 and 600 mg consecutively) once per menstrual cycle. Six of the nine women also received a dose of 200 mg. The 36 pregnant women were randomized into four groups which were given a single dose of 25, 100, 400 or 600 mg RU 486. Blood samples were taken up to 120 h after dosing. Peak concentrations of RU 486 occurred on most occasions within 2 h. Plasma concentrations at 1 h and at 24 h increased in proportion to log dose. There was a wide variability (up to ten fold) in the pharmacokinetic parameters within each dose group. Plasma concentrations of RU 42633 were similar to those of RU 486 but concentrations of RU 42848 and RU 42698 were much lower. As with RU 486, the plasma concentrations of the metabolites were maintained at high levels for up to 48-72 h after dosing. The findings were consistent with a rapid metabolism of RU 486 to RU 42633; removal of the second methyl group leading to RU 42698 occurred much more slowly and to a much less extent than removal of the first. There appeared to be no significant differences between the non-pregnant and pregnant women in either the plasma concentrations or pharmacokinetic parameters of RU 486 and its metabolites. PMID- 8403911 TI - Active oxygen species in copper intrauterine device users. AB - The mechanism of copper intrauterine device (Cu IUD) in limiting intrauterine infections is poorly understood. Copper ions may enhance the release of reactive oxygen species which are deleterious to the microbes. The present study compares the oxidative responses of adherent cell population of uterus prior to Cu-T insertion and at different post-insertion intervals. Increase in reactive oxygen intermediates was evident at 1 week post-insertion. However, the release of active oxygen species decreased thereafter. Further, these responses were only a local phenomenon as the peripheral blood monocytes failed to produce appreciable change following Cu-T insertion. Results suggest the protective role of active oxygen species in Cu IUD users which lasts for a brief period. The withering of respiratory burst activity later on may possibly prevent endometrial damage. PMID- 8403912 TI - Experiences of legal sterilization in Scandinavia. AB - The three Scandinavian countries--Denmark, Norway and Sweden--all legalized voluntary sterilization in the 1970s. Previous legislation had been very restrictive, limiting access to certain defined minority groups. During the two decades with this new possibility, sterilization has been accepted as a good contraceptive alternative, equally accessible for men and women above age 25 years. The sterilization frequency varies considerably between the three countries, being most popular in Norway and Denmark. The proportion of sterilization in males versus females is 40% in Denmark, 25% in Norway and 20% in Sweden. Published follow-up studies point out risk groups for regret of the intervention, and underline the need for good preoperative counseling. The overall results are very positive. PMID- 8403913 TI - Biological activity in the repopulating rat spermatocyte after the withdrawal of gossypol treatment. IV. The activity for the modification of core histones. AB - Mature male rats were treated with gossypol for 8 weeks. Afterwards, treatment was halted to allow the arrested spermatogonia to revive. Fifteen days after the withdrawal of the drug treatment, the repopulating pachytene spermatocyte (RPS) had a lower level of core histone H3 and H4 acetylation (40 and 55% reduction, respectively) than that of the control pachytene spermatocyte (CPS). The reduction in core histone acetylation was found in histone H3 and H4 but not in H2A and H2B. Forty-five days after the withdrawal of the drug treatment, the inhibitory effect on core histone acetylation was recovered. Both dot-blot and standard liquid assays were used to detect the nuclear histone acetylase activity in RPS and CPS. Fifteen days after the withdrawal of gossypol treatment, the acetylase type A activity in RPS was reduced by 42% when compared to CPS. It has been concluded that after gossypol treatment, the activity for nucleosomal core histones acetylation was selectively inhibited. This effect is related to an inhibitory effect on the histone acetylase activity. PMID- 8403914 TI - Antigestagenic activity of Ixora finlaysoniana in rat. AB - Oral administration of crude ethanolic extract of the serial parts of Ixora finlaysoniana Wall. ex G. Don to adult female rats at 250 mg/kg dose on days 1-5 or 1-7 post-coitum prevented pregnancy in 100% rats. The extract was also effective when administered on days 1 or 1-3 post-coitum, but the minimum effective dose increased with decreased duration of administration and was 1000 mg/kg and 500 mg/kg, respectively, in the two schedules. At lower doses, a significant reduction in implantation number and increased post-implantation resorption rate were observed in all the schedules. Almost complete resorption of all implantations was observed after administration of 1000 mg/kg dose of the extract during the peri-implantation period. A slight acceleration in tubal transport rate of embryos and delay in blastocyst formation were observed in rats treated postcoitally with the single anti-implantation dose of the extract. Significantly fewer embryos were recovered after their entry into the uterus. Except in one rat receiving 250 mg/kg dose of the extract on days 1-5, in which one apparently normal zona-free blastocyst was recovered from the uterus, uterine flushings of none of the nonpregnant animals contained any unimplanted embryos by day 10 post-coitum. In immature rat bioassay, the extract was found to possess estrogenic activity as evidenced by dose-dependent increase in uterine weight and cornification of the vaginal epithelium at doses ranging from 50-1000 mg/kg. At the 500 and 1000 mg/kg doses, it also induced premature opening of the vagina. Taking 100% increase in uterine weight as the parameter, the extract was found to be about 1.6X10(5) times less estrogenic than ethinylestradiol. The extent and duration of estrogenic responses exerted by single contraceptive dose of the extract were also markedly lower than that induced by ethinylestradiol. The extract was devoid of any estrogen antagonistic or synergistic activity and did not affect ovarian prenidatory estrogen or progesterone synthesis. The findings indicate that the extract at its contraceptive dose a) exerts a differential estrogenic response at the fallopian tube and the uterine levels, b) does not appear embryocidal, but causes slight asynchrony in development and tubal transport rate of pre-implantation embryos, which together with their loss through vagina after entry into the uterus, due to estrogenic action of the extract, might contribute to its anti-implantation action, and c) its anti implantation and post-implantation resorptive actions are not mediated via altered ovarian function. PMID- 8403915 TI - What we have learned from recent IUD studies: a researcher's perspective. AB - Many studies published on intrauterine devices (IUDs) during the last six years have consistently reported findings in favor of IUD use. Notable among these findings are: IUDs are not abortifacients; newly developed IUDs are highly effective and the efficacy is long-lasting; IUDs can be safely used by most lactating women, with lower removal rates attributable to bleeding and/or pain; and immediate postplacental IUD insertion reduces the risk of expulsion usually associated with postpartum insertion. Most importantly, in apparent contrast to results often reported in the late 1960s through the early 1980s, recent findings show that IUDs per se, especially the medicated ones, are not associated with an increased risk of pelvic inflammatory disease (PID), nor are they associated with an increased risk of ectopic pregnancy or subsequent infertility. There are still issues concerning IUD use that are controversial in spite of numerous studies. Should some of the contraindications currently listed for IUD use be modified according to the newer findings? Is the risk of uterine perforation increased when the IUD is inserted in lactating women? Do IUD tails increase the risk of PID? Does oral use of antibiotics at IUD insertion help prevent postinsertion PID? There are also issues that have not been sufficiently addressed and more information from empirical studies is needed. These include: the effect of the insertor's skill on IUD performance; IUD use in nulliparous as well as in older women; the relationship between IUD use and chlamydia infection; and long-term IUD use and safety, including actinomycosis, etc. Answers are also needed by administrators facing difficult programmatic decisions. For instance, should programs involving massive IUD removal be implemented as many IUD-wearing women are approaching or passing menopause? Similarly, are large programs to remove less-effective devices and replace them with newer and more effective IUDs advisable? This article reviews the state-of-the-art findings from recent IUD studies on the above issues. PMID- 8403916 TI - A time to be born: how the fetus signals to the mother that it is time to leave the uterus. PMID- 8403917 TI - Decreased incidence of postoperative urinary incontinence with a modified Penrose drain technique for treatment of prostatic abscesses in dogs. AB - Drainage with multiple Penrose drains is currently recommended as the primary treatment for canine prostatic abscesses. A recent report indicated that short term and long term urinary incontinence is a frequent complication associated with this technique. In this study, a modified multiple Penrose drain technique that avoids dissection dorsal to the prostate gland was performed in 17 dogs. Long term follow up information was obtained by telephone interview with the owners. Three dogs had urinary incontinence that resolved spontaneously within 3 days of surgery. Within the first year after that period, no other dogs developed incontinence. An excellent result was recorded in 12 dogs (71%) and a good result in 5 dogs (29%) having prostatic abscess drainage. PMID- 8403918 TI - Total protein and electrophoretic pattern of cerebrospinal fluid in sheep with some common neurological disorders. AB - Cerebrospinal fluid (CSF) samples were collected from normal sheep and cases of some common ovine neurological diseases and analysed by agarose gel electrophoresis. Protein fraction concentrations determined from clinically normal sheep of mixed ages were used to establish reference values. The CSF albumin and globulin concentrations were significantly increased in meningo encephalitis cases (P < 0.05) compared to healthy sheep. The group mean CSF albumin percentage for each of the neurological diseases studied was not significantly different from the mean CSF albumin concentration for control sheep (P > 0.05). In the ovine neurological diseases studied in this series, agarose gel electrophoresis of CSF did not add further to the information already gathered from the total CSF protein concentration. PMID- 8403919 TI - Diaphragmatic herniation causing respiratory signs in a heifer. AB - A diaphragmatic hernia was found in a pre-partum dairy heifer that was exhibiting signs of respiratory disease. Ruminal tympany was included in the history although this was not a feature of the disease on admission. Diagnosis was made by auscultation and confirmed by radiology and during elective Caesarean section to remove the fetus. PMID- 8403920 TI - Choroidal melanoma in a dog. AB - A 7-year-old male Great Dane dog was presented with blindness and a hemorrhagic retinal detachment in the right eye. Ocular ultrasonography revealed a conical shaped mass in the posterior segment that protruded into the vitreous cavity. Histologic examination of the enucleated eye revealed a heavily-pigmented melanocytoma arising from the choroid lateral to the optic disc. PMID- 8403921 TI - Repetitive injectable anesthesia in a 27-year-old horse. AB - A 27-year-old horse was anesthetized 3 times a week for 4 weeks, to facilitate cobalt therapy of a squamous cell carcinoma in the left paranasal sinus. Limitations of facilities required transport of the anesthetized horse to and from the cobalt therapy room, therefore, injectable anesthesia was used. Initially, the horse was preanesthetized with xylazine (at 1.1 mg/kg IV) and butorphanol (0.04 mg/kg IV). After 3 anesthetic episodes, the xylazine dose was reduced to 0.4 mg/kg IV and the butorphanol was deleted from the regimen. Tiletamine-zolazepam (1.1 mg/kg IV) was used for induction and maintenance of anesthesia (although on 2 occasions a bolus of ketamine was used to maintain immobility). The length of the procedure varied from 10-45 minutes, and the recovery times to sternal and standing varied from 23-76 and 55-105 minutes respectively. Although numerous complications might be expected given the age of this horse and the physiologic and metabolic demands of repetitive injectable anesthetics, none were observed. PMID- 8403922 TI - Polycystic hepatic disease, thoracic granular cell tumor and secondary hypertrophic osteopathy in a horse. AB - A 13-year-old American Saddlebred mare was presented with a 4-day history of anorexia. Physical examination revealed increased inspiratory effort and bony enlargement of the distal limbs. Radiographs indicated a thoracic mass and periosteal proliferations on the distal limbs consistent with hypertrophic osteopathy. Gastric endoscopy revealed distal esophageal and gastric ulceration, and functional pyloric stenosis. Abdominal ultrasonographic examination revealed multiple large, cystic structures associated with the liver. A percutaneous biopsy indicated the thoracic mass to be a granular cell tumor. At necropsy, a large mass consisting of intercommunicating cystic structures was present confluent with the right caudal edge of the liver. Histologically these hepatic lesions were consistent with cystic hepatic disease, which has not previously been reported in the horse. PMID- 8403923 TI - Persistent hyperbilirubinemia in a healthy thoroughbred horse. AB - Persistent hyperbilirubinemia and icterus are described in a healthy 4-year-old Thoroughbred horse. Hyperbilirubinemia was not related to food intake and was not associated with evidence of increased hemolysis or with acquired hepatic disease. The hyperbilirubinemia was thought to be a result of inappropriate conjugation of bilirubin rather than any abnormality in bilirubin uptake or excretion. The bilirubinemia in this horse appears most similar to a human syndrome, caused by a familial deficiency of bilirubin-uridine diphosphate glucuronyl transferase. PMID- 8403924 TI - The effect of roughage source on exercise performance and metabolism in thoroughbred horses. AB - The effects of roughage source on metabolism and exercise capacity were investigated using 6, previously conditioned, mature thoroughbred horses in a cross-over experiment. The horses were assigned to either non-alfalfa or alfalfa roughage diets which were isocaloric. The diets were fed for 2 weeks, after which the horses were exercise tested and then fed the alternate diet. Horses were exercised on a high-speed treadmill using a rapid incremental test. Arterial blood samples were collected for blood gas analysis and acid base measurements and venous blood samples for lactate and red blood cell potassium analysis. Mixed expired gas samples were collected for measurement of oxygen uptake, carbon dioxide production and respiratory exchange ratio. Heart rate was measured by telemetry ECG and respiratory frequency by observation. When the 2 diets were compared there were no significant differences in treadmill exercise capacity nor in any of the metabolic or cardiorespiratory measurements made before, during or after the exercise period. We concluded that roughage source has no effect on cardiorespiratory and metabolic function during or after exercise, and does not affect endurance performance during a treadmill exercise test. PMID- 8403925 TI - [The Cardiology Society yesterday and today]. PMID- 8403926 TI - [Remembrance of Prof. Kruta]. PMID- 8403927 TI - [Significance of transesophageal echocardiography in valvular and aortic diseases]. PMID- 8403928 TI - [Interesting topics from the angiology seminar held 18-8 March 1993 in Prague]. PMID- 8403929 TI - [Postoperative conditions--a new field in invasive cardio-angiology]. PMID- 8403930 TI - [First anniversary meeting of the Czech Cardiology Society (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. How to treat the patient with cardiac insufficiency]. PMID- 8403931 TI - [First anniversary meeting of the Czech Cardiology Society. (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. Heart transplantation]. PMID- 8403932 TI - [First anniversary meeting of the Czech Cardiology Society (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. Patients after surgery for congenital heart defects in adulthood]. PMID- 8403933 TI - [First anniversary meeting of the Czech Cardiology Society (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. Patients after correction of heart valve defects]. PMID- 8403934 TI - [First anniversary meeting of the Czech Cardiology Society (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. Patients with dual chamber pacing--DDD, VDD]. PMID- 8403935 TI - [First Anniversary Meeting of the Czech Cardiology Society (Zlin, 19-21 May 1993). Reports of coordinators of individual working groups. How to treat the patient with hypertension]. PMID- 8403936 TI - An almost forgotten anniversary. First International Congress of Cardiology (Prague, June 2 and 3, 1933) PMID- 8403937 TI - A three-site programmed electrical stimulation protocol in patients with and without ventricular tachyarrhythmias. AB - Programmed electrical stimulation (PES) of the ventricle plays an important role in diagnosis and in testing the effect of pharmacological and non-pharmacological therapy of ventricular tachyarrhythmias. A stimulation protocol should reliably reproduce clinical ventricular tachyarrhythmia. The authors compared the results of the standard one-site protocol to those of a new test delivering extrastimuli alternately into the right and left ventricle. In group I (control group, n = 37), clinically without ventricular tachycardia, tachycardia could not be induced in any of the patients. In group II (n = 30), with clinical ventricular tachycardia, inducibility was in 22 patients using the standard test, and in 23 patients using the new test. The authors conclude that this modification of the PES protocol with alternate extrastimulus delivery into the right and left ventricle does not seem to contribute significantly to PES sensitivity compared to the standard one-site stimulation protocol. It is not clear whether or not another stimulation protocol with alternate stimulus delivery to both ventricles or to different sites in the right ventricle will raise the sensitivity, specificity and reproducibility of the PES test. PMID- 8403938 TI - Endogenous quinidine-like immunoreactivity in the serum of rats with cardiac overload, stress or hyperthyroidism. Rapid communication. AB - Quinidine-like immunoreactivity (Abbott TDx Quinidine fluorescence polarization immunoassay) was measured in control, sham-operated, abdominal aorta stenotic and hyperthyroid (thyroxine feeding 0.1 mg/rat/day rats. The mean quinidine levels (mumol quinidine/1 serum) were 0.07, 0.14, 0.13 and 0.25, respectively. The elevation in experimental hyperthyroidism was statistically (analysis of variance, Duncan's test) significant. The possibility of the existence of an endogenous antiarrhythmic, immunologically cross-reacting in quinidine immunoassay, is discussed. PMID- 8403939 TI - [Myocarditis and endomyocardial biopsy]. AB - In a group of endomyocardial biopsies (EMB), performed in 45 patients with the clinical diagnosis of suspect myocarditis or dilated cardiomyopathy, positive findings (including histologically suspect lesions) were made in 53.3% of first EMB. Myocarditis, particularly its focal or multifocal forms, is bound to pose a major challenge not only in clinical diagnosis but, often, also in histological diagnosis. Of paramount importance in this situation are 1) early specimen removal after the onset of clinical manifestations and before initiation of the appropriate therapy, 2) removal of a major number of specimens in a single EMB because of the potential of focal lesions. PMID- 8403940 TI - [Atrial rate and increased work capacity during DDD pacing in patients with a high level atrioventricular block]. AB - Though the benefits of DDD pacing in patients with high-degree atrioventricular (AV) block are undisputable, it is desirable to identify--because of economic aspects--the subgroups of patients with maximum benefit from DDD pacing. An attempt has been made in the presented study to identify the subgroups of patients with a high increase in maximum working capacity during DDD pacing. The predictability of artificial pacemaker block based on atrial rate behaviour during exercise in heart block and VVI pacing has also been determined. The results are presented obtained in 26 patients with high-degree AV block and normal sinus function, who underwent exercise testing during AV block, VVI and DDD pacing. The benefit from DDD pacing in maximum working capacity was higher in patients with a lower resting atrial rate, steeper slope and a higher mean increase of atrial rate in the course of exercise during VVI pacing only. On the other hand, the maximum atrial rate during AV block is a good predictor of the incidence of artificial pacemaker block during exercise during DDD pacing and provides useful information for setting the upper rate limit of DDD pacemaker. PMID- 8403941 TI - [Transjugular intrahepatic portosystemic shunt]. AB - The transjugular intrahepatic portosystemic shunt (TIPS) is a relatively new method of creating a portosystemic shunt using a needle, angioplasty balloon catheters and expandable metallic stents. During a 6-month period, the authors have performed TIPS, using the spiral Z-stent--another modification of the Gianturco-Rosch stent--in 13 patients with portal hypertension. The procedure was technically successful in all patients. Portal pressure decreased by 6 mmHg in one group of patients with 9-10 mm stents, and by 12 mmHg in another group using 12 mm stents. Control of variceal bleeding or resolution of refractory ascites was evident in 11 of the 13 patients. PMID- 8403942 TI - [The effect of indobufen on aortocoronary bypass patency after 1 week and after 1 year]. AB - A randomized clinical study was conducted to compare the effect of "conventional" antiaggregation therapy (ASA plus dipyridamole) versus indobufen in patients after aortocoronary bypass (ACB) surgery. The patency of venous ACB using coronary DSA one week and one year after surgery was evaluated in 52 patients divided into two groups. The study included only ACB's with intraoperative blood flow rates < or = 40 ml/min as it is just these ACB's which are at the highest risk of early and late occlusions. While, in the ASA plus dipyridamole-treated group, occlusions were found in 11 of the 39 reconstructions (28.2%), the proportion was nine out of 37 procedures (24.3%) in the indobufen group. One year after surgery, occlusion was found in 14 out of 32 ACB's (43.7%) in the ASA plus dipyridamole group compared to 14 occlusions in 31 ACB's (45.2%) in the indobufen group. The difference in the number of occlusions between the two groups was not statistically significant. Because of some benefits of indobufen compared to ASA (shorter time of effect, superior tolerance in patients with ulceration), the former drug can be recommended for use in some indicated cases. PMID- 8403943 TI - [Drug therapy of chronic congestive heart failure: the why and how]. AB - As the prevalence of congestive heart failure is steadily increasing correct treatment becomes more and more important. In pathophysiology, clinical signs once believed to express the effort of the body to "compensate" for the failure (hypervolaemia, tachycardia, sympathetic stimulation) have been recognized as sort of a physiological error: to a primary signal which still remains not fully understood, the body responds in a similar way as to dehydration. As late as the mid-sixties, "function curves" represented a framework for our conceptual thinking and digitalis the essential treatment. Since then, we had to realize that the function of the failing ventricle is not determined just by filling and contractility-the way it is in the healthy ventricle is-but also by aortic impedance. Vasodilators have become the cornerstone of therapy. At present, diuretics are the most important drugs for congestive heart failure treatment. Thiazides, given together with loop diuretics, ameliorate extracellular fluid volume in all treated patients, but their effect is accompanied by potassium losses due to increased aldosterone levels. Angiotensin converting enzyme (ACE) inhibitors are unique in not only decreasing ventricular afterload and improving haemodynamics but, also, in decreasing circulating norepinephrine levels, protecting kidneys from inappropriate efferent arteriolo-constriction and suppression of aldosterone secretion. In other words, ACE inhibitors change the essential physiological set-up: they turn a high-perfusion-pressure body into a low-perfusion-pressure one. Furthermore, they convincingly improve life expectancy. Combination of diuretics and ACE inhibitors is the best we can do for our patients today. Although useful in some patients, the definite value of digitalis, and particularly its effect on longevity, remains an unresolved question. PMID- 8403944 TI - The practice of critical care--describing who we are, evaluating what we do, and computing the cost. PMID- 8403945 TI - No headlines, just headway. PMID- 8403946 TI - Selective digestive decontamination in critically ill patients. PMID- 8403947 TI - Atrial fibrillation following coronary artery bypass surgery: new hope for an old problem? PMID- 8403948 TI - Accurate estimation of glomerular filtration in the intensive care unit: another Holy Grail? PMID- 8403949 TI - Extracorporeal life support for pediatric respiratory failure: a remembrance of things past. PMID- 8403950 TI - Value and cost of teaching hospitals: a prospective, multicenter, inception cohort study. AB - OBJECTIVE: To examine variations in case-mix, structure, resource use, and outcome performance among teaching and nonteaching intensive care units (ICU). DESIGN: Prospective inception cohort study. PATIENTS: A consecutive sample of 15,297 patients at 35 hospitals, which compared 8,269 patients admitted to 20 teaching ICUs at 18 hospitals vs. 7,028 patients admitted to 17 non-teaching ICUs at 17 hospitals. INTERVENTIONS: None. MEASUREMENTS: We selected demographic, physiologic, and treatment information for an average of 415 patients at each ICU, and collected data on hospital and ICU structure. Outcomes were compared using ratios of observed to risk-adjusted predicted hospital death rates, ICU length of stay, and resource use. MAIN RESULTS: When compared to nonteaching ICUs, teaching ICUs had twice the number of physicians who regularly provided services and cared for significantly younger and more severely ill (p < .001) patients. Risk-adjusted ICU length of stay was similar, but resource use was significantly (p < .001) greater in teaching ICUs, with $3,000 (10.5%) of estimated total costs for an average ICU admission related to increased use of diagnostic testing and invasive procedures in teaching ICUs. Risk-adjusted hospital death rates were not significantly different (p = .1) between all teaching and nonteaching ICUs, but were significantly (p < .05) better in four teaching ICUs, but in only one nonteaching ICU. The 14 hospitals that were members of the Council of Teaching Hospitals had significantly better risk adjusted outcome in their 16 ICUs than all others (odds ratio = 1.21, confidence interval 1.06 to 1.38, p = .004). CONCLUSIONS: Teaching ICUs care for more complex patients in a substantially more complicated organizational setting. The best risk-adjusted survival rates occur at teaching ICUs, but production cost is higher in teaching units, secondary to increased testing and therapy. Teaching ICUs are also successfully transferring knowledge to trainees who, after their training, are achieving equivalent results at slightly lower cost in nonteaching ICUs. PMID- 8403951 TI - Improving intensive care: observations based on organizational case studies in nine intensive care units: a prospective, multicenter study. AB - OBJECTIVE: To examine organizational practices associated with higher and lower intensive care unit (ICU) outcome performance. DESIGN: Prospective multicenter study. Onsite organizational analysis; prospective inception cohort. SETTING: Nine ICUs (one medical, two surgical, six medical-surgical) at five teaching and four nonteaching hospitals. PARTICIPANTS: A sample of 3,672 ICU admissions; 316 nurses and 202 physicians. MATERIALS AND METHODS: Interviews and direct observations by a team of clinical and organizational researchers. Demographic, physiologic, and outcome data for an average of 408 admissions per ICU; and questionnaires on ICU structure and organization. The ratio of actual/predicted hospital death rate was used to measure ICU effectiveness; the ratio of actual/predicted length of ICU stay was used to assess efficiency. MEASUREMENTS AND MAIN RESULTS: ICUs with superior risk-adjusted survival could not be distinguished by structural and organizational questionnaires or by global judgment following on-site analysis. Superior organizational practices among these ICUs were related to a patient-centered culture, strong medical and nursing leadership, effective communication and coordination, and open, collaborative approaches to solving problems and managing conflict. CONCLUSIONS: The best and worst organizational practices found in this study can be used by ICU leaders as a checklist for improving ICU management. PMID- 8403952 TI - Eliminating needless testing in intensive care--an information-based team management approach. AB - OBJECTIVE: To determine if the application of an information-based management system in adult intensive care units (ICU) can produce sustained decreases in the use of laboratory resources and costs. DESIGN: Interventional study with prospective data collection on consecutive patients admitted during three time periods. SETTING: A 10-bed adult surgical ICU and an eight-bed adult medical ICU in a tertiary care hospital. PATIENTS: All patients admitted to an ICU during a 7 month baseline period (n = 647), a 1-yr intervention period (n = 1236), and a 2 yr follow-up period (n = 2349). INTERVENTIONS: Using a management database to track the use of 123 laboratory investigations during the baseline period, nine frequently ordered investigations (determination of blood gases, glucose, potassium, electrocardiogram, chest radiograph, sodium, chloride, complete blood count with differential, and serum osmolality) were targeted for reduction. Specific policies were developed by a multidisciplinary committee within the ICU to reduce the utilization of these laboratory, radiology, and cardiology tests. The policies were applied to all patients admitted during the 1-yr intervention period and during the 2-yr follow-up period. MEASUREMENTS AND MAIN RESULTS: A 25% reduction was observed in the frequency of all 123 monitored tests during the intervention period. The most dramatic reductions occurred in the nine targeted tests (range 19% to 46%) (p < .001). There were significant reductions in only 13 of the untargeted 114 investigations during this period. Potential annual cost savings were > $150,000 Canadian. No increases in ICU mortality rate, length of stay, or cost of medication were observed, and the reductions in the frequency of targeted tests were maintained during the 2-yr follow-up period. CONCLUSIONS: Application of an information-based multidisciplinary management system in the ICU can produce marked and sustained reductions in unnecessary testing in a cost effective manner. Although rationing of intensive care services may be necessary, reducing needless testing can be a safe and effective cost-containment strategy in the ICU. PMID- 8403953 TI - Glasgow Coma Scale score in the evaluation of outcome in the intensive care unit: findings from the Acute Physiology and Chronic Health Evaluation III study. AB - OBJECTIVE: To investigate the ability of the Glasgow Coma Scale score to predict hospital mortality rate for adult medical-surgical intensive care unit (ICU) patients without trauma. DESIGN: A prospective cohort analysis of adult medical surgical patients from a nationally representative sample of 40 U.S. hospitals. PATIENTS: 15,973 consecutive, nontraumatic ICU admissions and a comparison group of 687 head trauma admissions. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients' gender, age, treatment location before ICU admission, comorbidities, admission diagnosis, daily physiologic measurements, Glasgow Coma Scale score, Acute Physiology and Chronic Health Evaluation (APACHE III) score, subsequent hospital mortality rate, and unit-specific sedation practices were noted. Hospital mortality rates were stratified by the first ICU day Glasgow Coma Scale score for all admissions. The relationship between the Glasgow Coma Scale score and outcome for two high mortality medical diagnoses (post-cardiac arrest and sepsis) were also examined and compared to the relationship found in patients with head trauma. The Glasgow Coma Scale score on ICU admission had a highly significant (r2 = .922, p < .0001) but nonlinear relationship with subsequent outcome in ICU patients without trauma. Discrimination of patients into high- or low-risk prognostic groups was good, but discrimination in the intermediate levels (Glasgow Coma Scale score of 7 to 11) was reduced. This relationship varied within the operative and nonoperative groups, and also within different disease categories, various age groups, and certain ranges of the Glasgow Coma Scale score. A reduced initial Glasgow Coma Scale score associated with sepsis was a combination of factors associated with a higher mortality rate than that found in patients with head trauma. The proportion of patients who could not be assigned a Glasgow Coma Scale score because of sedation/paralysis varied widely across ICUs. The overall predictive capability of the APACHE III Prognostic Scoring System was improved by incorporating the Glasgow Coma Scale score. CONCLUSIONS: We demonstrated the prognostic importance of admission levels of consciousness as measured by the Glasgow Coma Scale score on ICU and hospital mortality rates. We concluded that the Glasgow Coma Scale score may be used to stratify and predict mortality risk in general intensive care patients, but lack of sensitivity in the intermediate range of Glasgow Coma Scale Score should be noted. Ideally, the Glasgow Coma Scale score should also be applied in the context of other physiologic information and the patient's specific diagnosis. Variation in the use of sedatives in different ICUs means that imputing or substituting a value other than normal for an unobtainable Glasgow Coma Scale score may introduce a substantial treatment bias into subsequent outcome predictions. PMID- 8403954 TI - Selective decontamination of the digestive tract in neurosurgical intensive care unit patients: a double-blind, randomized, placebo-controlled study. AB - OBJECTIVES: The aim of this study was to assess, in a selected population, the effects of selective decontamination of the digestive tract on colonization of the oropharynx, trachea, stomach and rectum, and on the infection rate. An economical assessment was also performed. DESIGN: A prospective, double-blind, randomized, placebo-controlled, dual-center trial. SETTING: Two neurosurgical intensive care units. PATIENTS: A total of 191 comatose patients admitted emergently and intubated within < 24 hrs were enrolled. Of these patients, 68 were excluded because they either died, got an early infection, or were extubated within the first 5 days. A total of 123 patients were analyzed: 63 treated and 60 placebo patients. INTERVENTIONS: Topical antibiotics (tobramycin, polymyxin E, amphotericin B) were applied in the oropharynx and in the stomach. Vancomycin was added in the oropharyngeal paste. Placebo patients received the same regimen (i.e., a suspension of fluid and a paste) but without antibiotics. No parenteral antibiotics were given during the study period. MEASUREMENTS AND MAIN RESULTS: Bronchopneumonia episodes were diagnosed with protected specimen brush or plugged telescoping catheter and other infections were diagnosed according to the Center for Disease Control of Atlanta criteria. Antibiotic costs and cost per survivor were calculated. Selective decontamination of the digestive tract significantly reduced Gram-negative bacilli colonization as well as the number of episodes of bronchopneumonia, urinary tract infections, and sinusitis. Despite the addition of vancomycin, Staphylococcus aureus remained the main potential pathogen causing tracheal colonization and subsequent bronchopneumonia. The reduction in bronchopneumonia rate was observed in head-trauma patients only. We were able to show that: a) the trachea was the main reservoir of microorganisms responsible for pneumonia; b) pneumonia developed after tracheal colonization. Total charges for antibiotics were 2.8 times higher in the treated group than in the placebo group; in calculating the cost per survivor, selective decontamination of the digestive tract might be beneficial due to the reduced length of stay. CONCLUSIONS: Selective decontamination of the digestive tract is an effective technique in reducing infectious morbidity in comatose neurosurgical patients. Because of its cost, this technique should be used only in selected populations. PMID- 8403955 TI - Prophylactic procainamide for prevention of atrial fibrillation after coronary artery bypass grafting: a prospective, double-blind, randomized, placebo controlled pilot study. AB - OBJECTIVE: To evaluate the effect of prophylactic procainamide on the frequency of postoperative atrial fibrillation in patients undergoing myocardial revascularization. DESIGN: Prospective, randomized, double-blind, placebo controlled pilot study. SETTING: Surgical intensive care unit and wards at a university hospital affiliate. PATIENTS: A total of 46 patients undergoing myocardial revascularization. INTERVENTIONS: Twenty-two patients received procainamide (procainamide group) and 24 patients received placebo (control group). Procainamide was administered to the procainamide group within 1 hr of the patient's arrival in the intensive care unit and consisted of an intravenous loading dose (12 mg/kg) followed by a maintenance dose (2 mg/min) of procainamide. The control group received a similar volume of placebo. When the patient was able to take oral medication, the study drug was administered orally in a weight-adjusted dosage. MEASUREMENTS: Electrocardiograms (EKGs) were continuously monitored. Procainamide and N-acetyl procainamide serum concentrations were measured, and the dosages in the procainamide group were adjusted by an independent observer. The study drug was continued for 5 days or until an event occurred that resulted in dismissal from the study. MAIN RESULTS: The procainamide group and control group had similar preoperative demographic descriptors and operative variables, except for the mean left ventricular ejection fraction, which was lower in the control group than in the procainamide group (60% vs. 68%, p = .03 [Wilcoxon rank-sum test]). There were no hospital deaths. The number of episodes of postoperative atrial fibrillation was significantly reduced in the procainamide group (5 episodes in 129 patient days at risk [3.9%/day at risk]) compared with the control group (17 episodes in 161 patient days at risk [10.6%/day at risk], p = .04 [Fisher's exact test]). Complication rates were similar in both groups. CONCLUSIONS: In a pilot trial, prophylactic procainamide reduced the number of episodes of atrial fibrillation in patients after coronary artery bypass grafting. Procainamide also decreased the number of patients who experienced postoperative atrial fibrillation. However, due to the small sample size, this latter difference was not statistically significant. Further studies are needed to confirm this encouraging trend. PMID- 8403956 TI - A prospective, randomized study of continuous versus intermittent nebulized albuterol for severe status asthmaticus in children. AB - OBJECTIVES: a) to determine if continuous nebulization of albuterol is more effective than intermittent nebulization in the treatment of children with status asthmaticus and impending respiratory failure; b) to determine the effect of continuous nebulization and intermittent nebulization on duration of hospital stay and bedside respiratory therapy care. DESIGN: Prospective, randomized study. SETTING: A pediatric intensive care unit (ICU) in a university children's hospital. PATIENTS: Seventeen pediatric asthmatic patients with severe status asthmaticus, with impending respiratory failure (Woods asthma score > or = 5), and without evidence of cardiac or other preexisting lung disease were admitted to the pediatric ICU. The patients were randomized to receive continuous nebulization (n = 9) or intermittent nebulization (n = 8) of albuterol. INTERVENTIONS: The asthmatic patients were randomized into two groups. The continuous group received 0.3 mg/kg/hr of albuterol by continuous nebulization; the intermittent group received 0.3 mg/kg of albuterol over 20 mins every hour. All patients received aerosol therapy through the same delivery system. MEASUREMENTS: Responses to therapy were evaluated by following asthma score, arterial blood gas values, hemodynamics, FIO2, and arterial oxygen saturation initially and at 30 mins, 1 hr, 2 hrs, 4 hrs, then every 4 hrs for a 24-hr time period. Patients were determined to no longer be in impending respiratory failure when their asthma score was < 5 for four consecutive hours. Electrocardiograms, serum electrolyte values, and creatine phosphokinase total and MB fraction values were obtained before and after treatment. Hospital stay and respiratory therapy time, in relative value units (one relative value unit = 10 mins) were analyzed from data collected from therapist bedside flow sheets. RESULTS: The patient characteristics (demographics, hemodynamics, arterial blood gas values, asthma severity, corticosteroid use, theophylline, and beta 2-adrenergic receptor agonist administration) before entry into the study did not differ between groups. Patients in the continuous group improved more rapidly and were out of impending respiratory failure sooner than patients in the intermittent group (continuous group = 12 (median) hrs (range 4 to 24) vs. intermittent group = 18 (median) hrs (range 12 to 24; p = .03). Bedside respiratory therapy time evaluated by relative value units was less for patients who received continuous nebulization of albuterol (continuous group = 14 (median) relative value units (range 6 to 26) vs. intermittent group = 33 (median) relative value units (range 25 to 49; p = .001). Hospital stay was shorter for patients who received continuous nebulization of albuterol (continuous group = 80 [median] hrs [range 51 to 173] vs. intermittent group = 147 [median] hrs [range 95 to 256]; p = .043). Hemodynamics, serum potassium, and creatine phosphokinase concentrations did not differ before and after the study in either group. CONCLUSIONS: In children with impending respiratory failure due to status asthmaticus, continuous nebulization of albuterol is safe and results in more rapid clinical improvement than intermittent nebulization. Respiratory therapy required at the bedside and duration of hospital stay were substantially less for patients receiving continuous nebulization of albuterol, which suggests that continuous nebulization of albuterol is more cost effective than intermittent nebulization. PMID- 8403957 TI - Predictability of creatinine clearance estimates in critically ill patients. AB - OBJECTIVES: a) To evaluate the predictive ability of different creatinine clearance methods as compared with the criterion standard, inulin clearance; and b) to determine which of the predictive methods yields the most accurate estimation of creatinine clearance. DESIGN: Prospective study. SETTING: Medical intensive care unit (ICU) of a university-affiliated tertiary care hospital. INTERVENTIONS: Glomerular filtration rate was measured by the criterion standard, inulin clearance. PATIENTS: Twenty mechanically ventilated adults. MEASUREMENTS: Renal function was assessed by the following procedures: inulin clearance using a standard protocol, 30-min creatinine clearance, 24-hr creatinine clearance, and creatinine clearance estimates by the Cockcroft-Gault equation. Ideal body weight, total body weight or lean body mass with actual serum creatinine or serum creatinine concentration corrected to 1 mg/dL (85 mumol/L) in cachectic patients were sequentially incorporated into the Cockcroft-Gault equation. RESULTS: The Cockcroft-Gault equation, using ideal body weight and the corrected serum creatinine concentration, was the best predictor of inulin clearance with the smallest bias (9.7 +/- 8.6, 95% confidence interval 5.7 to 13.8). The bias encountered with the 30-min creatinine clearance was not different from that value with the 24-hr creatinine clearance (21.6 +/- 33.0, 95% confidence interval 6.2 to 37.1 vs. 25.4 +/- 28.3, 95% confidence interval 11.8 to 42.9). Good correlations existed between inulin clearance and the Cockcroft-Gault equation, using ideal body weight and the corrected serum creatinine concentration (r2 = .81; p = .0001), as well as between inulin clearance and the Cockcroft-Gault equation, using the lower of ideal or total body weight and the higher of the actual serum creatinine concentration or corrected serum creatinine (r2 = .75; p = .0001). The 30-min creatinine clearance and the 24-hr creatinine clearance had poorer agreement with inulin clearance. The incorporation of a corrected serum creatinine value into the Cockcroft-Gault equation consistently led to better predictions and higher correlation coefficients. CONCLUSIONS: The utilization of the Cockcroft-Gault equation as used clinically (the lower of ideal or total body weight and the higher of actual serum creatinine or corrected serum creatinine concentration to 1 mg/dL [85 mumol/L]) results in more accurate predictions of glomerular filtration rate in the medical, critically ill patient than urine creatinine clearance measures. If creatinine clearance measures are used, the 30 min collection provided results not different from those results obtained with 24 hr urinary collections. PMID- 8403958 TI - Decreased polymorphonuclear leukocyte exudation in critically ill anergic patients associated with increased adhesion receptor expression. AB - OBJECTIVE: To determine the mechanism for the reduced polymorphonuclear leukocyte exudation in critically ill anergic patients. DESIGN: Prospective consecutive patient study. SETTING: Tertiary care surgical intensive care unit. PATIENTS: Eighteen patients with intra-abdominal injections were studied. INTERVENTIONS: Critically ill patients were stratified based on their delayed type hypersensitivity response to ubiquitous antigens. Polymorphonuclear leukocytes were isolated from blood and from exudate blister type skin windows. Adhesion and chemotactic surface receptors were measured, as was cytokine content and chemoattraction capacity of skin window fluid for control neutrophils. MEASUREMENTS AND MAIN RESULTS: Circulating neutrophils from anergic patients have increased CR3 adherence receptors compared with those neutrophils from reactive patients. f-met-leu-phe receptors are equal in number and C5a receptors are either significantly reduced in number or occupied with ligand. This same receptor pattern is maintained after neutrophil exudation in both patient groups. Serum and skin window fluid from anergic patients attracted less neutrophils in vitro and in vivo. CONCLUSIONS: These data suggest two possible mechanisms for the reduced neutrophil delivery of critically ill anergic patients: a receptor mediated increased adherence to vascular endothelium preventing diapedesis; reduced chemo-attraction potential of serum, and possibly, exudate fluid. PMID- 8403959 TI - Effects of norepinephrine, epinephrine, and dopamine infusions on oxygen consumption in volunteers. AB - OBJECTIVE: To determine the relationships between plasma concentrations of norepinephrine, epinephrine, and dopamine and oxygen consumption (VO2) during infusion of these catecholamines. DESIGN: Prospective, randomized variable dose, pharmacologic study in which a noncumulative infusion-rate design was used. SETTING: Laboratory of the Department of Anesthesiology at a University Hospital. PATIENTS: Twenty-one normal volunteers. INTERVENTIONS: After a control period of 20 mins, norepinephrine (three infusion rates; 0.06 to 0.2 microgram/kg/min; n = 7), epinephrine (four infusion rates; 0.02 to 0.2 microgram/kg/min; n = 7), or dopamine (three infusion rates; 3 to 12 micrograms/kg/min; n = 7) was administered to normal volunteers (n = 21) for the purpose of constructing plasma concentration/VO2 response curves. MEASUREMENTS AND MAIN RESULTS: Systolic and diastolic blood pressure, heart rate, plasma concentrations of norepinephrine, epinephrine, and dopamine, and VO2 were measured at the end of the control period and at the end of each catecholamine infusion. VO2 was measured using a ventilated canopy system and a differential oxygen sensor. Typical hemodynamic responses to vasopressors were seen during adrenergic receptor agonist infusions. VO2 increased from 132 +/- 7 to 153 +/- 10 mL/min/m2 during the highest infusion rate of norepinephrine, from 133 +/- 7 to 182 +/- 11 mL/min/m2 during the highest infusion rate of epinephrine, and from 132 +/- 13 to 163 +/- 8 mL/min/m2 during the highest infusion rate of dopamine (p < .05; paired t-test). Increases in VO2 were correlated with the logarithms of the corresponding plasma catecholamine concentrations. Effects on VO2 and hemodynamic responses occurred at similar plasma concentrations for each of the three catecholamines. CONCLUSIONS: Administration of norepinephrine, epinephrine, or dopamine results in marked increases in VO2 in volunteers. In patients, the administration of catecholamines or sympathomimetics to attain optimal values of cardiac index, oxygen delivery (DO2), and VO2 may increase the oxygen demand and thus obscure the DO2-VO2 relationship. PMID- 8403960 TI - Titration of intravenous anesthetics for cardioversion: a comparison of propofol, methohexital, and midazolam. AB - OBJECTIVE: To compare propofol, methohexital, and midazolam administered as titrated infusions for sedation during electrical cardioversion. DESIGN: A prospective, randomized, single-blind comparative study. SETTING: Coronary care unit in a military teaching hospital. PATIENTS: Thirty adult patients with atrial fibrillation, atrial flutter, or paroxysmal supraventricular tachycardia. Each patient required electrical cardioversion. Patients were randomized to receive one of the three study drugs. Ten patients composed one drug group. MEASUREMENTS AND MAIN RESULTS: Demographic variables were similar between groups. Patients were randomized to receive propofol (10 mg/mL), methohexital (5 mg/mL), or midazolam (0.5 mg/mL) administered at 10 mL/min until the patients failed to follow verbal commands and demonstrated a degradation of the lid response to stimulation. Dose requirements (mean +/- SD) were propofol 1.69 +/- 0.46 mg/kg, methohexital 1.07 +/- 0.34 mg/kg, and midazolam 0.16 +/- 0.06 mg/kg. Hemodynamic assessment at baseline, after induction, after cardioversion, and at recovery demonstrated no difference in mean arterial pressure between the three groups. The time to awakening was significantly prolonged in the group that received midazolam (33 +/- 11 mins, p < .05) as compared with the times of the groups that received propofol (11 +/- 4 mins) and methohexital (9 +/- 3 min). Side-effects were similar between groups, with the exception of an increase in pain on injection with propofol and an increased frequency of confusion in those patients receiving midazolam. Recall of the electrical discharges at one hour after the procedure occurred in two patients in the propofol group. In both cases, there were technical problems which caused the duration of the procedure to extend into the anticipated recovery period. Unit dose costs at our institution for a 70-kg patient are: methohexitol, $3.14 (500-mg bottle); medazolam, $14.88 (5-mg vials x 3); and propofol, $6.60 (200-mg ampule). CONCLUSIONS: All three drugs are acceptable choices for use during elective direct-current cardioversion. Titration of the agent results in a total drug dose which is usually less than the typical induction dose. There were no significant differences in the hemodynamic actions of these drugs at any time interval. Both propofol and methohexital proved superior in their ability to provide a more rapid anesthetic onset and recovery as compared with midazolam. Propofol offers the advantage of requiring no premixing or dilution, and it is not a controlled substance, although it does result in more pain on injection. PMID- 8403961 TI - Effect of barbiturate therapy on phenytoin pharmacokinetics. AB - OBJECTIVE: To evaluate the effect of high-dose pentobarbital therapy on phenytoin pharmacokinetics. DESIGN: A prospective, clinical study. SETTING: The intensive care unit of a university hospital. PATIENTS: Ten adult patients with cerebral lesions requiring anticonvulsants and control of intracranial pressure. INTERVENTIONS: Each patient received phenytoin sufficient to maintain a plasma concentration at 15 micrograms/mL (60 mumol/L) both before and after barbiturate therapy. Plasma concentrations of total phenytoin, unbound phenytoin, and the major metabolite of phenytoin, 5-(p-hydroxyphenyl)-5-phenylhydantoin, were measured, and pharmacokinetic variables obtained before and after barbiturate therapy were compared. MEASUREMENTS AND MAIN RESULTS: Plasma concentrations of total phenytoin remained within the therapeutic range during the 12-hr period preceding barbiturate therapy. After barbiturate therapy, plasma concentrations of both total and unbound phenytoin were significantly less than those concentrations before barbiturate therapy. For total phenytoin, maximum metabolic velocity was increased by 62% (1.09 +/- 0.62 to 1.77 +/- 0.52 mg/L/hr, 1.20 +/- 0.68 to 1.95 +/- 0.57 nmol/L/sec, p < .05), and area under the plasma concentration-time curve (0 to infinity) and mean residence time were each decreased by 73% (32.5 +/- 20.0 to 8.7 +/- 3.1 min.mg/mL, 2.14 +/- 1.25 to 0.57 +/- 0.19 sec.mmol/L, p < .01, and 135,000 +/- 69,000 to 37,000 +/- 11,000 secs, p < .005, respectively) after barbiturate therapy. The plasma concentration of the principal metabolite of phenytoin, 5-(p-hydroxyphenyl)-5-phenylhydantoin, was significantly increased after barbiturate therapy. CONCLUSIONS: Phenytoin metabolism is increased by barbiturate therapy, and supplemental doses of phenytoin and frequent drug monitoring may be required after barbiturate therapy. PMID- 8403962 TI - Lack of agreement between measurement of ejection fraction by impedance cardiography versus radionuclide ventriculography. AB - OBJECTIVE: To determine the limits of agreement between left ventricular ejection fraction estimated using systolic time intervals from impedance cardiography and left ventricular ejection fraction estimated by radionuclide ventriculography. DESIGN: A prospective study for diagnostic tests using radionuclide ventriculography as the criterion standard. SETTING: A large military teaching hospital. PATIENTS: Twenty ambulatory adults scheduled for radionuclide ventriculography. MEASUREMENTS AND MAIN RESULTS: A regression equation to estimate ejection fraction from systolic time intervals is available in a widely used impedance-based cardiac monitoring device. The estimated ejection fraction is then used in an equation with stroke volume estimated by the same device to calculate an end-diastolic volume. We studied the agreement of the ejection fraction as estimated by this device with the ejection fraction estimated by radionuclide ventriculography by obtaining simultaneous estimates of ejection fraction over a broad range of adult patients. Twenty ejection fraction pairs were analyzed. The correlation of ejection fraction by impedance cardiography to ejection fraction by radionuclide ventriculography was significant (r2 = .55; p < .002). However, the mean difference between the technologies was -8.85%, with a standard deviation of the differences of 7.15%, resulting in a 95% confidence range for agreement of -23.2% to +5.5%. CONCLUSIONS: The 95% confidence range defining the limits of agreement between ejection fraction by impedance cardiography and ejection fraction by radionuclide ventriculography is not clinically acceptable. In the opinion of the authors impedance cardiography should not be used in place of radionuclide ventriculography for the assessment of ejection fraction at this time. PMID- 8403963 TI - Comparison of ear-based, bladder, oral, and axillary methods for core temperature measurement. AB - OBJECTIVE: To determine the accuracy and repeatability of ear-based, bladder, oral, and axillary temperature methods. DESIGN: Prospective, descriptive comparison of the accuracy of four temperature methods in relation to pulmonary artery temperature and the repeatability of each method. SETTING: Critical care units of a university teaching hospital. PATIENTS: Convenience sample of 38 adult patients with indwelling pulmonary artery thermistor catheters. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Ear-based estimates of core temperature with an infrared thermometer and pulmonary artery, bladder, oral, and axillary temperatures with thermistor-based instruments were made every 20 mins for 4 hrs. Mean offsets (+/- SD) from pulmonary artery temperature for each method were as follows: ear-based 0.07 +/- 0.41 degrees C; bladder 0.03 +/- 0.23 degrees C; oral 0.05 +/- 0.26 degrees C; and axillary -0.68 +/- 0.57 degrees C. The accuracy of each method varied with the level of pulmonary artery temperature. Repeated measurements with all four methods had mean SD values within +/- 0.2 degrees C. CONCLUSIONS: Infrared ear thermometry provided a relatively close estimate of pulmonary artery core temperature, although with more variability than bladder or oral methods, while axillary readings were substantially lower than the pulmonary artery temperature and highly variable. PMID- 8403964 TI - Accuracy of a new bedside method for estimation of circulating blood volume. AB - OBJECTIVE: To evaluate the accuracy of a modification of the carbon monoxide method of estimating the circulating blood volume. DESIGN: Prospective, clinical study comparing two methods for estimating the circulating blood volume. Carbon monoxide kinetics were studied by computer simulation. SETTING: Intensive care unit in a university hospital. PATIENTS: Intensive care patients receiving intermittent positive-pressure ventilation and healthy volunteers. MEASUREMENTS AND MAIN RESULTS: The circulating blood volume was determined from measurements of the carbon monoxide saturation of hemoglobin before and after rebreathing a gas mixture containing 20 to 50 mL of carbon monoxide for a period of 10 to 15 mins. A specially designed Water's "to & fro" system was developed to avoid any leakage during intermittent positive-pressure ventilation. Blood samples were taken before, during, and immediately after rebreathing. The amount of carbon monoxide administrated during each rebreathing maneuver resulted in an average increase in the carbon monoxide saturation of hemoglobin of 3.6%. The washin of carbon monoxide was completed within the first 4 to 6 mins of the rebreathing period in nine healthy subjects and within 10 mins in 12 intensive care patients. The 95% limits of agreement between the carbon monoxide method and the radioactive isotope-labeling method was +/- 540 mL (+/- 2 SD). CONCLUSION: Determination of circulating blood volume can be performed with sufficient accuracy using an amount of carbon monoxide that gives rise to an unharmful increase in the carboxyhemoglobin concentration. PMID- 8403965 TI - Convulsions and hypertension in children: differentiating cause from effect. AB - OBJECTIVE: To determine if the magnitude of blood pressure (BP) increase could differentiate convulsion caused by a hypertensive crisis from a primary convulsive disorder, which itself increases BP. DESIGN: Retrospective analysis. SETTING: Admission to a pediatric intensive care unit (ICU) within a tertiary care center. PATIENTS: All children with hypertensive crisis admitted to the pediatric ICU from 1976 to 1990 were studied. Thirty-eight episodes occurred in 36 patients. The charts of children admitted for status epilepticus from 1976 to 1986 were also reviewed. One hundred and fifty-three episodes occurred in 145 patients. MEASUREMENTS AND MAIN RESULTS: BP values at entry to the pediatric ICU in patients with hypertensive crisis were compared with the highest BP values obtained within an hour after cessation of convulsion in 120 patients admitted for status epilepticus. The Z scores for BP, adjusted for age and sex, were compared. The BP values for children in hypertensive crisis with or without convulsions were by far greater than the BP values observed in patients in status epilepticus (p < .0001). For a patient in the postictal phase, a BP > or = 4.0 SD above the mean for age and sex predicted with 78% probability the presence of a hypertensive crisis requiring emergency treatment. If the BP was < 4 SD below the mean, the possibility of a hypertensive crisis was excluded (negative-predictive value 100%). CONCLUSIONS: Children with hypertensive crisis, as well as children with status epilepticus, can present with a high BP. In a postictal patient, the magnitude of BP increase is a useful clinical parameter to exclude a hypertensive crisis that requires specific treatment of the BP. PMID- 8403966 TI - A case-control study of patients readmitted to the intensive care unit. AB - OBJECTIVE: To determine characteristics of patients requiring readmission to an intensive care unit (ICU). DESIGN: Retrospective case-control chart review. SETTING: Adult patients in an 8-bed medical and 16-bed surgical ICU in a 650-bed university teaching hospital during an 18-month period. INTERVENTIONS: None. METHODS: Each patient readmitted to either ICU was compared to a single control patient matched for age, sex, unit, and diagnosis. A total of 117 variables were obtained from chart review. The last value for each variable before ICU discharge was used. Chi-square analysis, linear regression factor analysis, correlation, and Student's t-test were used to identify significant factors predicting the patient's return to an ICU. Descriptive statistics were used to describe various subgroups. RESULTS: Mortality rate was 41.5% in the 82 readmitted patients (comprising 4.6% of the total ICU admissions) and only 7.3% in the controls. Mean (+/- SD) hospital length of stay was 47.8 +/- 42.0 days in the study group and 20.8 +/- 14.2 days (p < .0009) in the controls. Initial ICU length of stay was 8.3 +/- 16.1 days in the readmitted group and 4.0 +/- 5.0 days (p = .02) in the controls. Variables that predicted readmission to the ICU were: increased respiratory rate (24.2 vs. 20 breaths/min, p < .002), lower hematocrit value (31.9% vs. 34.4%, p = .01), positive fluid balance (p < .03), and positive blood cultures (six in the readmitted group, none in the controls, p = .002). More than 30% of readmissions were for a recurrence or worsening of the original problem. Fifty-four percent of the patients were readmitted with pulmonary failure. Mortality rate in both groups was related to advanced age, Acute Physiology and Chronic Health Evaluation (APACHE II) scores at ICU discharge, and increased blood urea nitrogen concentrations. Mortality rate in the control group, but not in the readmitted group, correlated with the level of consciousness, serum creatinine concentration, and use of hemodialysis. CONCLUSIONS: Readmission to an ICU carries a risk of high mortality rate and increased length of stay and may represent premature discharge in at least 30% of patients. Pulmonary failure is the immediate cause of readmission in more than half of the readmitted patients. Increased respiratory rate correlates with ICU readmission. Intermediate care areas for patients with poor pulmonary function may help to avoid readmission to an ICU, prevent death, and conserve hospital resources. PMID- 8403967 TI - Hypertonic saline infusion in hemorrhagic shock treated by military antishock trousers (MAST) in awake sheep. AB - OBJECTIVES: To determine the effect of military antishock trousers (MAST) combined with hypertonic saline in controlled hemorrhagic shock in an awake sheep model. DESIGN: Prospective, randomized, controlled study. SETTING: University research laboratory. SUBJECTS: Healthy adult sheep (21 to 33.5 kg) were studied in four experimental groups. INTERVENTIONS: Sheep underwent cannulation of the jugular vein and the carotid artery. An indwelling catheter was also inserted into their bladders. Mean arterial, pulmonary arterial, pulmonary artery occlusion, and central venous pressures, cardiac and stroke volume indices, systemic and pulmonary vascular resistances, PaO2 and PaCO2, and serum lactate and hemoglobin concentrations were measured at baseline, after induction of hemorrhagic shock, and subsequently during resuscitative treatments of normal saline, MAST, hypertonic saline, and the combination of MAST and hypertonic saline. MEASUREMENTS: Controlled hemorrhagic shock was induced by arterial bleeding of 40 mL/kg in all animals. The sheep were randomized into four groups. In group 1 (n = 5), controlled hemorrhagic shock was treated by 5 mL/kg sodium chloride 0.9% (isotonic saline). In group 2 (n = 6), controlled hemorrhagic shock was treated by 5 mL/kg sodium chloride 7.5% (hypertonic saline). In group 3 (n = 5), controlled hemorrhagic shock was treated by MAST inflated to 40 mm Hg followed by 5 mL/kg sodium chloride 0.9%. In group 4, controlled hemorrhagic shock was treated by MAST (40 mm Hg) followed by 5 mL/kg of hypertonic saline. MAIN RESULTS: Arterial bleeding was followed by significant decreases in mean arterial pressure (87 +/- 1 to 41 +/- 4 mm Hg; p < .001), cardiac index (4.6 +/- 0.4 to 2.0 +/- 0.2 L/min/m2; p < .001), and urine output (102 +/- 30 to 13.0 +/- 7 mL/hr; p < .001), and an increase in systemic vascular resistance (1517 +/- 130 to 2601 +/- 370 dyne.sec/cm5; p < .001). MAST inflation in group 3 increased systemic vascular resistance to 3018 +/- 399 dyne.sec/cm5 (p < .05) and mean arterial pressure to 79 +/- 5 mm Hg (p < .05), while cardiac index, urine output, and lactate concentration remained unchanged. Infusion of hypertonic saline after MAST inflation (group 4) resulted in an increase in mean arterial pressure to 99 +/- 4 mm Hg (p < .001), an increase in cardiac index to 4.1 +/- 0.4 L/min/m2 (p < .001), an increase in urine output to 221 +/- 93 mL/hr (p < .001), and a decrease in systemic vascular resistance to 1847 +/- 175 dyne.sec/cm5 (p < .005). CONCLUSIONS: In hemorrhagic shock, the combination of MAST and hypertonic saline increases mean arterial pressure, improves cardiac output and tissue perfusion during the application of MAST, and also prolongs for > 2 hrs the short beneficial effect of hypertonic saline on mean arterial pressure. PMID- 8403968 TI - Effect of phenytoin on acute lung injuries in unanesthetized sheep. AB - OBJECTIVE: To determine if the intravenous administration of phenytoin attenuates or prevents acute experimental lung injury. DESIGN: Placebo-controlled, longitudinal animal investigative study. SETTING: University research laboratory. SUBJECTS: Sixteen yearling female lambs weighing 30 +/- 3 kg. INTERVENTION: After administration of anesthesia, the animals were endotracheally intubated and mechanically ventilated. Using sterile techniques, four thoracotomies were performed. Through the left fourth intercostal space, cannulas for pressure measurements were inserted directly into the main pulmonary artery and left atrium. An ultrasound flow cuff for determination of cardiac output was placed around the main pulmonary artery. Through the left tenth intercostal space, the diaphragmatic and mediastinal parietal pleura were widely cauterized. Through the right tenth intercostal space, the caudal mediastinal lymph node was identified and divided at the caudal margin of the right pulmonary ligament, and a 1- to 2 cm portion of the node distal to the ligament was resected. The diaphragmatic and mediastinal parietal pleura were widely cauterized. Through the right sixth intercostal space, the efferent duct (or ducts) was identified, ligated at the site of entry into the thoracic duct, and cannulated. The lymph cannula was brought to the outside of the thorax through a separate stab wound. MEASUREMENTS AND MAIN RESULTS: Unanesthetized sheep were studied 7 to 10 days after surgery. Hemodynamic, lung fluid balance, and arterial blood variables were measured in uninjured sheep and in sheep injured by intravenous infusions of Escherichia coli endotoxin (1 microgram/kg iv over 30 mins), air bubbles (0.056 to 0.074 mL/kg/min over 4 hrs), or oleic acid (0.06 mL/kg over 1 hr). The sheep were studied when untreated and after pretreatment with phenytoin. We found that the expected increase in protein-rich lung lymph flow with injuries, resulting from increased microvascular permeability in the lungs, was attenuated by phenytoin when the lungs were injured by endotoxin or air bubbles. In contrast, phenytoin had no effect on oleic acid-induced lung injury or on uninjured lungs. CONCLUSIONS: Phenytoin attenuates acute lung injuries in sheep that are thought to be caused by stimulation of host inflammatory responses (e.g., endotoxin and air bubbles), but has no effect on direct injuries to the lungs (e.g., oleic acid). A plausible mechanism for this finding is phenytoin inhibition of polymorphonuclear leukocyte function. PMID- 8403969 TI - End-tidal CO2 changes under constant cardiac output during cardiopulmonary resuscitation. AB - OBJECTIVES: To evaluate a) whether end-tidal CO2 values change under constant cardiac output during cardiopulmonary resuscitation (CPR), and b) what factors are responsible for the change. DESIGN: A cohort study. SETTING: University research laboratory. SUBJECTS: Nine mongrel dogs. INTERVENTIONS: Ventricular fibrillation was electrically induced. After 2 mins, open-chest cardiac massage was initiated to maintain cardiac output at 0.2 L/min (23% of baseline cardiac output) by the measurement of blood flow with an electromagnetic flow probe on the ascending aorta. The cardiac massage was kept constant until 50 mins after the induction of ventricular fibrillation. MEASUREMENTS AND MAIN RESULTS: Before and during ventricular fibrillation, end-tidal CO2, minute volume of alveolar ventilation, and CO2 excretion were continuously monitored. Blood gases and oxygen saturation values were also measured in arterial and the mixed venous blood samples. CO2 content was calculated. After induction of ventricular fibrillation, end-tidal CO2 decreased and thereafter continued to increase until the end of the experiment. Two mechanisms may have contributed to the early reduction in end-tidal CO2. One mechanism is a further decrease in CO2 excretion compared with the reduction in alveolar ventilation and the other is an increase in alveolar deadspace (estimated from the increase in the difference between PaCO2 and end-tidal CO2). The subsequent increase in end-tidal CO2 was mainly due to a change in CO2 excretion. There are two hypotheses concerning the subsequent increase in CO2 excretion: the increase in pulmonary capillary blood flow (estimated from the change in the arteriovenous CO2 content gradient) and the increase in CO2 production itself. CONCLUSIONS: End-tidal CO2 changes under constant cardiac output during CPR. When end-tidal CO2 is used to estimate the effectiveness of the cardiac massage, this type of change must be recognized. PMID- 8403970 TI - Antifibrillatory and electrophysiologic actions of moricizine alone and in combination with lidocaine: a prospective, randomized trial. AB - OBJECTIVE: The Cardiac Arrhythmia Suppression Trial II showed that moricizine acutely increases the occurrence of sudden cardiac death. Thus the objective of this investigation was to evaluate the antifibrillatory properties of moricizine (a new antiarrhythmic agent) alone and in combination with lidocaine (an established antifibrillatory agent). DESIGN: Prospective, double-blind, randomized, placebo-controlled trial. SETTING: Laboratory at a large, university affiliated medical center. SUBJECTS: Eighteen domestic farm swine with a mean weight of 39 +/- 5 kg. INTERVENTIONS: After pentobarbital anesthesia, the animals were instrumented. A bipolar pacing catheter was placed in the right ventricular apex and a pig-tail catheter was placed in the aortic arch for induction of ventricular fibrillation and aortic blood pressure monitoring. Subsequently, the pigs were randomized to moricizine or control (0.9% saline) groups. Each group underwent three treatment phases: baseline, drug (moricizine 2 mg/kg loading dose, 1.5 mg/kg/hr infusion, or saline bolus and infusion), and drug combined with lidocaine (5 mg/kg loading dose, 4 mg/kg/hr infusion). Ventricular fibrillation threshold was determined every 5 to 10 mins over a 1-hr period during each treatment phase. RESULTS: Ventricular fibrillation threshold values in the animals randomized to control were 16.8 +/- 7.6, 18.1 +/- 8.9, and 23.9 +/ 10.4 mA at baseline during saline infusion, and when saline was combined with lidocaine, respectively. The values during the saline-lidocaine combination treatment phase were significantly greater than the values at baseline and during saline treatment alone (p < .001). Ventricular fibrillation threshold values in the animals randomized to receive moricizine were 15.5 +/- 4.4, 18.1 +/- 5.1, and 21.1 +/- 8.4 mA at baseline, during moricizine infusion, and when moricizine was combined with lidocaine. The values during the lidocaine-moricizine combination treatment phase were significantly greater than values at baseline (p = .005), but not during moricizine treatment alone (p = .16). The increase in ventricular fibrillation threshold from baseline to moricizine (17%) was similar to the increase from baseline to saline (7%), p = .37. The increase in ventricular fibrillation threshold when lidocaine was added to moricizine (13%) was less than the increase with lidocaine alone (32%), p = .05. CONCLUSION: In this experimental model, moricizine, at the dose studied, lacked antifibrillatory properties. Moreover, moricizine did not contribute to the antifibrillatory effects of lidocaine. PMID- 8403971 TI - Reinstitution of digoxin after digoxin Fab antibody therapy in a hemodialyzed patient. PMID- 8403972 TI - "Snare loop" technique utilizing a wire basket for removing intravascular foreign bodies. PMID- 8403973 TI - Intracerebral hemorrhage: pathophysiology and management. AB - OBJECTIVES: To review the pathophysiologic basis for intensive care management of patients with intracerebral hematoma and to present management strategies based on that analysis. DATA SOURCES: Review of English language scientific and clinical literature using BRS Colleague search. STUDY SELECTION: Pertinent literature was selected to demonstrate physiologic principles, the clinical presentation, and the natural history of intracerebral hematoma. All clinical treatment trials were reviewed. DATA EXTRACTION: Literature was reviewed to summarize physiologic principles and the results of clinical trials. The suggested medical management is based on the results of clinical trials and the applications of pathophysiologic principles. DATA SYNTHESIS: Spontaneous intracerebral hematomas acutely increase intracranial volume and reduce intracranial compliance, thus potentially increasing intracranial pressure and reducing cerebral perfusion. Therefore, any physiologic perturbation that could increase cerebral blood volume (hypercarbia, hypoxia, vasodilation) or reduce cerebral perfusion (hypotension) should be avoided. Acute hypertension may be a response to increased intracranial pressure and should be treated conservatively. Treatment of increased intracranial pressure is accomplished initially with hyperventilation, but this approach should be replaced rapidly with cerebrospinal fluid drainage alone or in combination with osmotic therapy employing diuretics, osmotic agents, or hypertonic saline to produce a state of hyperosmolality and euvolemia. Although the role of surgery remains controversial, it may be helpful in selected patients. Careful attention to pulmonary and metabolic status and to the consequences of therapy is required. Outcome is primarily determined by hemorrhage size, location, and initial level of consciousness. Stereotaxic aspiration of hematomas is a promising new therapy. CONCLUSIONS: The primary injury from intracerebral hemorrhage is exacerbated by disturbed intracranial physiology. Management should include principles that improve intracranial compliance and reduce intracranial pressure. PMID- 8403974 TI - Extracorporeal life support for pediatric respiratory failure: predictors of survival from 220 patients. AB - OBJECTIVE: The purpose of this report was to examine the Extracorporeal Life Support Organization registry database for predictors of outcome for severe pediatric respiratory failure managed with extracorporeal life support. DESIGN: Retrospective cohort study. SETTING: Extracorporeal Life Support Organization data registry. PATIENTS: All nonneonatal pediatric patients who were treated in the United States with extracorporeal life support for severe pediatric respiratory failure reported to the Extracorporeal Life Support Organization registry as of August 1991. Patients with congenital heart disease and congenital gastrointestinal malformations were excluded from study. INTERVENTIONS: Venoarterial or venovenous extracorporeal life support for severe life threatening pulmonary failures. MEASUREMENTS AND MAIN RESULTS: As of August 1991, 220 pediatric patients meeting study entry criteria were reported to the Registry having received extracorporeal life support for severe pulmonary failure, since 1982. Forty-six percent (102 of 220 patients) were successfully managed with this technology and survived to hospital discharge. The mean patient age was 36.8 +/- 51.6 months. Fifty-one percent of the patients were male. The mean duration of mechanical ventilation before extracorporeal life support was 6.3 +/- 5.9 days. Mean blood gas and ventilatory measurements obtained before extracorporeal life support were as follows: PaCO2 52 +/- 23 torr (6.9 +/- 3.0 kPa); PaO2 59 +/- 32 torr (7.8 +/- 4.3 kPa); estimated alveolar-arterial oxygen gradient 561 +/- 63.4 torr (74.8 +/- 8.5 kPa); peak airway pressure 49.5 +/- 13.1 cm H2O; mean airway pressure 24.3 +/- 8.2 cm H2O; positive end-expiratory pressure 11.8 +/- 6.3 cm H2O; ventilator rate 58 +/- 64.4 breaths/min; and FIO2 0.98 +/- 0.07. The mean duration of extracorporeal life support for all patients was 247 +/- 164 hrs. For the 102 survivors, the mean time for decannulation from extracorporeal life support to extubation from mechanical ventilation was 6.5 +/- 7.6 days. Stepwise multivariate logistic regression modeling found the following variables to be associated with patient survival: a) patient age, b) days of mechanical ventilation before extracorporeal life support, c) peak inspiratory pressure, d) alveolar-arterial oxygen gradient, and e) extracorporeal life support administered since December 31, 1988 (all p < .05). CONCLUSIONS: Extracorporeal life support may represent an effective rescue therapy for some selected pediatric patients with severe respiratory failure for whom conventional mechanical ventilation support has failed to improve. Predictors of survival for this life-support therapy exist that may be helpful for individual patient prognostication and future prospective study. PMID- 8403975 TI - Sepsis and organ failure definitions and guidelines. PMID- 8403976 TI - Relative humidity with heating wire. PMID- 8403977 TI - Caloric requirements for critically ill surgical patients. PMID- 8403978 TI - Blood vitamin concentrations during the acute-phase response. PMID- 8403979 TI - Tumor necrosis factor: an updated review of its biology. AB - OBJECTIVE: To review the role of tumor necrosis factor (TNF) in the pathogenesis of the septic shock syndrome. DATA SOURCES: The international English language literature from 1985 to present formed the basis for this review. MEDLINE was used to identify pertinent animal and human studies pertaining to the clinically relevant aspects of TNF and related cytokines. STUDY SELECTION: Those studies that focused on developments that may lead to advances in the therapy for septic shock syndrome were emphasized. Investigations that described in vivo and human results served as the primary database. DATA EXTRACTION: Animal studies were selected based on the similarity of the model pathogenesis and outcomes to the human clinical sepsis syndrome. Patient studies were selected on the basis of study design and sample size. DATA SYNTHESIS: The normal role of TNF and pathologic effects consequent to the excessive production of TNF in response to an overwhelming infection or injury are reviewed. Evidence establishes the role of TNF in septic shock syndrome. Novel therapies, such as anti-TNF monoclonal antibodies, soluble TNF receptors, or soluble TNF receptor-immunoglobulin G heavy chain fusion proteins, may confer protection against septic shock syndrome. CONCLUSIONS: TNF plays a major role in the pathogenesis of the septic shock syndrome. TNF exerts a range of beneficial and injurious effects that may ultimately lead to organ dysfunction and death. The burst of TNF release after endotoxemia promotes the progression of the shock syndrome even in the absence of further TNF release. New therapies targeted to the attenuation of TNF may hold promise for the management of patients with septic shock syndrome. PMID- 8403980 TI - Tumor necrosis factor in the pathogenesis of infectious diseases. AB - OBJECTIVES: To review the immunologic role of the cytokines and the specific role that tumor necrosis factor (TNF) plays in response to infection. The influence of bacterial lipopolysaccharide on TNF, the cytokine cascade, and resultant pathologies are also reviewed. DATA SOURCES: A MEDLINE search of the international English language literature from 1960 to the present was reviewed, but data from the past 5 yrs primarily formed the basis for this review. STUDY SELECTION: Those studies detailing the interaction of lipopolysaccharide, TNF, and other cytokines, and their roles in combating infection were emphasized. Investigations that described animal and human results served as the primary database. DATA EXTRACTION: Animal studies were selected based on the relevance of the model to the pathogenesis of the human clinical syndrome. Where they provided supportive evidence, patient studies were selected on the basis of study design. DATA SYNTHESIS: TNF plays a key role in the normal immune response to infection, limiting the spread of pathogens. Exaggerated physiologic responses occur under the influence of high concentrations of TNF that are released in response to overwhelming infection, resulting in aberrations in coagulation, cell adhesion, chemotaxis/transmigration, and vascular integrity. These pathologic effects may be inhibited by anti-TNF monoclonal antibodies and recombinant soluble receptor inhibitory proteins. CONCLUSIONS: TNF exerts both physiologic and pathologic effects in response to infection; these events may lead to organ dysfunction and death. Anti-TNF therapies appear to attenuate the injurious effects of TNF. PMID- 8403981 TI - Tumor necrosis factor and the therapeutic potential of anti-tumor necrosis factor antibodies. AB - OBJECTIVE: To review the relationship of tumor necrosis factor (TNF) to clinical sepsis and the clinical potential of anti-TNF therapy in decreasing morbidity and mortality rates due to sepsis. DATA SOURCES: The international English language literature was reviewed, including animal studies and human clinical trials regarding TNF, anticytokine therapy, and sepsis. STUDY SELECTION: Studies which characterized the immunopharmacologic interactions between TNF and sepsis were emphasized. DATA EXTRACTION: This study specifically focused on experiments and clinical trials that directly involve the activity of TNF or anti-TNF antibodies, particularly but not limited to data derived from septic patients. DATA SYNTHESIS: The relationship between TNF and sepsis is described. Clinical aspects of anti-TNF therapy (timing, empiric use) are discussed. Phase I, II, and III trail of anti-TNF antibodies in clinical trials are reviewed. CONCLUSIONS: Current clinical strategies for sepsis therapy are only partially effective. Recent immunopharmacologic advancements have resulted in the identification of TNF as a pivotal proinflammatory cytokine mediator of sepsis. Animal studies demonstrate that anti-TNF therapy protects animals from the morbidity and mortality of sepsis. Phase I clinical studies of anti-TNF antibodies demonstrate the safety of monoclonal antibody therapy. The therapeutic application of anti TNF antibodies in sepsis trials is ongoing. PMID- 8403982 TI - Preclinical review of anti-tumor necrosis factor monoclonal antibodies. AB - OBJECTIVE: To review the preclinical evidence for the role of tumor necrosis factor (TNF) in the pathogenesis of septic shock and to assess the preclinical efficacy of anti-TNF therapies for this clinical problem. DATA SOURCES: The international English language literature from 1986 to the present formed the basis of this review. MEDLINE was used to identify pertinent in vitro and animal studies pertaining to the pathobiology of TNF and the use of anti-TNF therapies, with special emphasis on antibody approaches. STUDY SELECTION: Those studies that focused on the mechanisms of action of TNF, its role in the inflammatory cascade, and the potential uses of anti-TNF therapies were emphasized. Investigations that described animal and human results served as the primary database. DATA EXTRACTION: Animal studies were selected based on the relevance of the model to the pathogenesis of the human clinical sepsis syndrome. Where they provided supportive evidence, patient studies were selected on the basis of study design. DATA SYNTHESIS: The administration of anti-TNF antibodies in baboons, monkeys, and other species that were administered lethal doses of bacteria or endotoxin suggest that this approach may limit organ damage and decrease the mortality rate caused by the septic shock syndrome. Therapy with anti-TNF monoclonal antibodies is reviewed. CONCLUSIONS: Bacterial challenge induces the release of TNF (among other mediators), which exerts both physiologic and toxic effects that may ultimately lead to organ dysfunction and death. New anti-TNF therapies such as anti-TNF antibodies appear to attenuate the injurious effects of TNF and promote survival in otherwise lethal septic shock animal models, suggesting a similar benefit might be obtained in the treatment of human septic shock. PMID- 8403983 TI - Role of tumor necrosis factor-alpha in disease states and inflammation. AB - OBJECTIVE: To review the animal and human data defining the role of tumor necrosis factor (TNF) in the pathogenesis of the septic shock syndrome, the systemic inflammatory response syndrome, and related pathologic states. DATA SOURCES: The international English language literature from 1975 to present formed the basis for this review. MEDLINE was used to identify pertinent animal and human studies. STUDY SELECTION: Those animal and human studies that focused on the mechanisms of action of TNF, its role in the inflammatory cytokine network, and the potential uses of anti-TNF therapies were emphasized. DATA EXTRACTION: Animal studies were selected based on the relevance of the model to the pathogenesis of the human systemic inflammatory response syndrome. Where they provided supportive evidence, human studies were selected on the basis of study design. DATA SYNTHESIS: TNF plays a major role in systemic inflammatory response syndrome secondary to infection, burns, trauma or hemorrhagic shock, and pancreatitis. TNF influences the outcome of other infectious processes, including allograft rejection, ischemia-reperfusion injury, delayed-type hypersensitivity, and granuloma development. The administration of anti-TNF antibodies, soluble TNF receptors, and related fusion proteins may limit organ damage and decrease mortality rate. CONCLUSIONS: Although the regulated release of TNF may exert normal physiologic effects, the uncontrolled production of TNF may lead to organ dysfunction and death. TNF mediates a variety of other physiologic processes that are unrelated to sepsis syndrome. New anti-TNF therapies appear to attenuate the injurious actions of TNF. PMID- 8403984 TI - Yeast sugar transporters. AB - Transport of sugars is a fundamental property of all eukaryotic cells. Of particular importance is the uptake of glucose, a preferred carbon and energy source. The rate of glucose utilization in yeast is often dictated by the activity and concentration of glucose transporters in the plasma membrane. Given the importance of transport as a site of control of glycolytic flux, the regulation of glucose transporters is necessarily complex. The molecular analysis of these transporters in Saccharomyces has revealed the existence of a multigene family of sugar carriers. Recent data have raised the question of the actual role of all of these proteins in sugar catabolism, as some appear to be lowly expressed, and point mutations of these genes may confer pleiotropic phenotypes, inconsistent with a simple role as catabolic transporters. The transporters themselves appear to be intimately involved in the process of sensing glucose, a model for which there is growing support. PMID- 8403985 TI - Photochemical reactions in organized and semiorganized media. AB - The article describes organic photochemical reactions in heterogeneous fields. The first part of the article includes an introduction of miscellaneous electrostatic fields adsorbing photoactive species and the second part summarizes the types of photochemical reactions in their fields. Photochemical reactions carried out in various heterogeneous fields, inorganic as well as organic, were classified by their reaction type, that is, unimolecular reactions, bimolecular reactions, energy transfer reactions, and electron transfer reactions. PMID- 8403986 TI - Induction of tolerance to freeze-thaw (FT) damage in mammalian cells by pre-FT hypothermia treatment. AB - Attached asynchronous exponential phase V79 Chinese hamster cells were pretreated by hypothermic cycling in Hepes growth medium by Method 3 (48 h at 25 degrees C + trypsin) or Method 4 (48 h at 25 degrees C + 3 h at 37 degrees C + trypsin) prior to a freeze-thaw (FT) cycle in Hepes growth medium. Pretreatment by Method 3 or 4 increased the FT survival by a factor of 3.4. This implies that the 3 h at 37 degrees C, after the 25 degrees C exposure, is not necessary to confer resistance to the subsequent FT cycle in the case of V79 cells. However, with RIF-1 mouse cells, the 3 h at 37 degrees C confers increased resistance. The increase in FT survival of V79 cells after the above pretreatments cannot be accounted for by changes in cell cycle age distribution. No heat shock proteins are produced by this pretreatment. Since pretreatment by Method 3 or 4 also makes the cells resistant to hyperthermia, three other pretreatments, making the cells thermotolerant, were tried. None of these pretreatments resulted in a change in FT survival of the cells. Interaction analysis of FT data, when pretreatment by Method 4 is combined with the presence of DMSO during the FT cycle, indicates that the pretreatment and DMSO act synergistically whether exponential or stationary phase cells are used. Furthermore, the pretreatments and L-glutamine also act synergistically. These pretreatments also increase the FT survival of the RIF-1 mouse cell line; again, pretreatment and DMSO act synergistically. Hence, the method is not limited to cells of hibernating mammals. PMID- 8403987 TI - Subzero nonfreezing storage of the mammalian cardiac explant. I. Methanol, ethanol, ethylene glycol, and propylene glycol as colligative cryoprotectants. AB - We employed hyperosmotic concentrations of penetrating cryoprotective agents (CPA) to store the isolated rat hearts unfrozen at subzero temperatures. The effect of acute exposure to CPA was assessed by flushing the hearts with CP-14, a cardioplegic solution, containing methanol (MeOH), ethanol (EtOH), ethylene glycol (EG), or propylene glycol (PG) for 2 min and reperfusing immediately with Krebs-Henseleit buffer in a working-heart model. The maximal doses that did not cause irreversible suppression of heart function were: MeOH, 1.78 M; EtOH, 1.27 M; EG, 0.84 M; and PG, 0.87 M. For nonfreezing storage, the hearts were flushed with CP-14 containing the highest tolerable concentrations of MeOH, EtOH, EG, or PG, stored for 6 h at -3.7, -2.8, and -1.4 degrees C, respectively, and then reperfused. Control cardiac output (CO) was 76.2 +/- 1.8 ml/min. Post reperfusional recovery of CO was 86% in MeOH hearts, 82% in EtOH hearts, 76% in EG hearts, and 79% in PG hearts. Thus MeOH offered not only the least cardiac suppressing effect but the lowest nonfreezing storage temperature. When storage time was extended, recovery and myocardial ATP level decreased with time in hearts flushed with CP-14 + 1.78 M MeOH and stored at -3.7 degrees C. The decay of function was faster than the decay of ATP level, suggesting energy was better preserved than function. The low return of function, however, may be related to CPA toxicity, osmotic stress, and ischemia/reperfusion injury. Nonfreezing storage at subzero temperatures using these CPAs may provide a novel approach to long-term cardiac preservation. PMID- 8403988 TI - Morphological and functional alterations in endothelium, smooth muscle, and nerve fibers in rabbit aorta after storage at 4 degrees C. AB - Structural and implied functional changes in endothelial cells (EC), smooth muscle cells (SMC), and nerve fibers (NF) in rabbit aorta were studied after storage in cold Krebs solution. Rings from the thoracic aorta were stored for 2, 4, 6, and 8 days in a refrigerator at 4 degrees C. Subsequently they were examined after 6 h in an organ bath at 37 degrees C and processed for transmission electron microscopy. The earliest and most prominent structural changes were observed in NF and varicosities. These changed after 2 days and were completely destroyed after 6 days. Less remarkable changes in both the intensity and the timeliness of alterations were found in EC. The majority of them showed changes after 4 days. After 8 days they were seriously destroyed, but still formed a relatively continuous layer. The most resistant cells were SMC, showing only slight alterations during the whole period. Contraction of aortic rings in response to transmural nerve stimulation (8 Hz) gradually decreased with the length of cold storage. After 4 days contraction was about 10% of the control value, while after 6 days it was entirely abolished. Maximal relaxation of noradrenaline-precontracted rings in response to acetylcholine was decreased to 46% after 4 days of storage, while after 8 days it was only 15% of the control value. The contractile response of aorta to exogenous noradrenaline did not change during the whole period except for Day 4 when the response was significantly increased. These results showed the different susceptibilities of the EC, the SMC and the NF of the thoracic aorta to cold storage at 4 degrees C (NF > EC > SMC). The observed structural changes were in close agreement with their functional expressions. PMID- 8403989 TI - A metabolic view of receptor activation in cultured cells following cryopreservation. AB - The effect of cryopreservation on agonist-induced receptor activation in mammalian cells was investigated with the Cytosensor microphysiometer, a biosensor that monitors cellular metabolic activity by measuring changes in extracellular pH. In this study, two different cell types--nonadherent TF-1 cells (from a human erythroleukemia patient) and adherent WT3 cells (CHO-K1 cells transfected with the m1 muscarinic acetylcholine receptor)--were cryopreserved by freezing in a disposable cell capsule used in the microphysiometer. The recovery of metabolic activity by TF-1 cells was observed over approximately 1 h following thawing. Responses of the TF-1 cells to granulocyte-macrophage colony-stimulating factor (GM-CSF) and platelet activating factor (PAF) were measured before cryopreservation and 90 min after thawing. The GM-CSF and PAF responses retained 71 +/- 14% and 73 +/- 10% of maximum stimulation, respectively. Post-thaw cholinergic stimulation of WT3 cells was 73 +/- 9% of its level in similarly treated but unfrozen cells. Cryopreservation caused no detectable difference in desensitization of the response due to repeated application of carbachol. These results demonstrate the feasibility of pharmacological studies with cryopreserved cells in the microphysiometer and further suggest that the microphysiometer may be useful in exploring the biological consequences of cryopreservation in the early post-thaw period. PMID- 8403990 TI - Corneal cryopreservation with chondroitin sulfate. AB - In an experimental study using porcine corneas we investigated the influence of several variables of freeze-thaw trauma on survival of corneal endothelial cells after corneal cryopreservation employing chondroitin sulfate as a cryoprotectant. We examined the influence of: (1) the concentration of chondroitin sulfate in the cryopreservation medium, (2) the concentration of fetal calf serum in the cryopreservation medium, (3) the cooling rate, and (4) the preincubation period in the cryopreservation medium before freezing. Controls consisted of corneas cryopreserved in culture medium without cryoprotectants and corneas frozen in dimethyl sulfoxide (Me2SO) by the method of Capella et al. (Preservation of viable corneal tissue. Cryobiology, 2, 116-121, 1965). Morphological evaluation was performed by determining endothelial cell density after staining with alizarin red S and trypan blue. Morphological evaluation was not performed directly after thawing but after a subsequent storage period in organ culture at 31 degrees C in order to detect latent cell damage after freeze-thaw trauma. The group that yielded the best endothelial cell density was evaluated in perfusion chamber experiments in order to measure the functional integrity of the tissue. Controls included corneoscleral rims from freshly slaughtered pigs, corneas stored in organ culture for 1 day, and fresh corneas mechanically denuded of endothelium. It was demonstrated that corneas that had been cryopreserved in MEM medium containing 2% chondroitin sulfate and 20% fetal calf serum with a cooling rate of 1 degree C/min displayed the highest endothelial cell density (2430 cells/mm2, SO = 383, n = 15) compared with freshly dissected corneas (3395 cells/mm2, SD = 200, n = 48). Control corneas frozen by the method of Capella et al. demonstrated only a poor outcome. We conclude that in our experimental environment, corneal cryopreservation with chondroitin sulfate provides higher corneal endothelial cell densities than corneas preserved by conventional methods. Moreover, even slight changes in variables that affect the tissue in the freeze-thaw cycle have a major impact on corneal endothelial cell survival after cryopreservation. PMID- 8403991 TI - Cryopreservation of rat islets of Langerhans: a comparison of two techniques. AB - Although a number of different protocols have been described for the cryopreservation of pancreatic islets of Langerhans, most involve equilibration with 2 M Me2SO followed by slow cooling and fast warming. This paper compares two methods with identical equilibration protocols: one with cooling at 0.25 degree C/min to -40 degrees C prior to transferring to liquid nitrogen and subsequent warming at 200 degrees C/min and sucrose dilution of Me2SO (Method A) and the other with cooling at 0.3 degree C/min to -70 degrees C prior to transferring to liquid nitrogen and warming at 50 degrees C/min followed by stepwise dilution of Me2SO (Method B). Islet viability was assessed by syngeneic transplantation to the renal subcapsular space of streptozotocin-induced diabetic rats. Seven hundred fifty fresh islets in the size range 100-200 microns consistently reversed diabetes (blood glucose < 10 mmol/liter) whereas 1000 Method A cryopreserved islets and 2000 Method B cryopreserved islets were required. Cryopreservation by either method resulted in impaired islet function compared to that of fresh islets, although this functional loss could be partially compensated by the transplantation of greater numbers of cryopreserved islets. PMID- 8403992 TI - Cryopreservation of porcine blastocysts by vitrification. AB - The objective of this study was to assess the possibility of cryopreservation of porcine expanded and hatched blastocysts by vitrification. The following four types of vitrification solutions were applied in this study: (i) EFT, a mixture of 7.2 M ethylene glycol, 0.003 M ficoll, and 0.3 M trehalose; (ii) DAP213, 2 M dimethyl sulfoxide, 1 M acetamide, and 3 M propylene glycol; (iii) DAP213-T, DAP213 supplemented with 0.3 M trehalose; and (iv) EPT, 4 M ethylene glycol, 3.1 M propylene glycol and 0.3 M trehalose. The embryos collected on Days 5 to 7 (Day 0 = onset of estrus) were allocated to eight experimental groups according to the types of vitrification solution and the developmental stage. In the toxicity test, the embryos were equilibrated in the respective vitrification solutions either in a single step or in four steps and then transferred to 1 M sucrose in a single step at 22 degrees C without cooling. As a whole, the viabilities of embryos equilibrated in the solutions were lower than those of control embryos. The stepwise equilibration was superior to a single-step equilibration in the in vitro survival of, especially, expanded blastocysts after dilution. No significant difference was observed between the vitrification solutions for the four-step method. In the vitrification test, the embryos were equilibrated in the solutions by the four-step method, loaded into a 0.25-ml plastic straw, and plunged into liquid nitrogen. Although viable embryos were obtained after warming from all of the combinations except the hatched blastocyst-EPT group, viabilities were further reduced by cooling (range of reduction rates: 60 to 100%). The possible cause of low survival after warming is also discussed concerning the cryophysical properties of vitrification solutions. PMID- 8403993 TI - Effect of sucrose, trehalose, hypotaurine, taurine, and blood serum on survival of frozen bull sperm. AB - Factorially arranged experiments were conducted to study the effects of adding sucrose and trehalose, known to have cryoprotective properties, blood serum, and the antioxidants taurine and hypotaurine on sperm motility after freezing and thawing at different rates. At sugar concentrations of 0.1 M, osmolality of the whole milk (WM) or egg yolk-Tris freezing medium (without glycerol) was about 370 mOsmol. Survival following freezing and thawing was reduced unless osmolality was corrected by adding pure water to reduce osmolality to about 280 mOsmol. Then the post-thaw percentages of motile sperm for control WM, and WM with 0.05 or 0.10 M sucrose, or 0.05 or 0.10 M trehalose, respectively, were 62, 55, 61, 57, and 62%. Thawing semen at 5 degrees C, versus 25 degrees C resulted in 64 versus 70% motile sperm (P < 0.05). Post-thaw survival of sperm stored at 25 degrees C for 24 h in trehalose-treated WM was superior to that in WM (P < 0.05). Hypotaurine and taurine had little effect on sperm survival. Up to 10% (v/v) of heated blood serum was generally beneficial, but gave more variable responses with different bulls. Sperm survival after cooling at -25 degrees C/min was slightly superior to that after cooling at -15 degrees C/min to -100 degrees C. The effects of the compounds studied on motility of frozen-thawed sperm were small, but if they protect sperm cell membranes, as reported for other types of membranes, they may assist sperm in surviving in the reproductive tract of the cow prior to fertilization. PMID- 8403994 TI - Survival of trophoblastic fragments and vesicles after vitrification, ultrarapid freezing, and storage at 4 degrees C. AB - Embryos recovered from superovulated cows on Day 13 or Day 17 of their sexual cycles were cut into fragments to make trophoblastic vesicles. The fragments were suspended in a cryoprotectant solution consisting of glycerol and sucrose and frozen by direct immersion into liquid nitrogen: alternatively, they were vitrified using the Massip method in a mixture of glycerol and 1,2-propanediol. Trophoblasts and fragments of a chilled embryo were placed into a temperature controlled humidified chamber for culture. Vesicles which developed from trophoblastic fragments were subjected to vitrification according to the method of Massip. We observed good survival of trophoblastic fragments which had been subjected to chilling, freezing, or vitrification. Their survival did not differ from the survival of "fresh" trophoblastic fragments. PMID- 8403995 TI - Challenges in critical care nursing: helping patients and families cope. PMID- 8403996 TI - Edith Sacks--after 60-plus years in nursing, retirement is not on her agenda. Interview by Michael Villaire. PMID- 8403997 TI - Are new graduates hired in your unit? If not, why not? What advice would you give them? PMID- 8403998 TI - Nurses as victims of violence. PMID- 8403999 TI - A dangerous new trend--the invisible nurse. PMID- 8404000 TI - Congratulations and clarification, please. PMID- 8404001 TI - Alternatives to food coloring in enteral tube feedings. PMID- 8404002 TI - The impact of nutrition on wound healing. PMID- 8404003 TI - Liver failure: case study of a complex problem. PMID- 8404004 TI - A conceptual framework for predicting selected complications of ICU patients. PMID- 8404005 TI - Transesophageal echocardiography: a new window on the heart and great vessels. AB - TEE adds an exciting new dimension to the field of echocardiography. It has expanded the physician's ability to clearly examine cardiac structures and bloodflow; in many instances it has been the means by which rapid and lifesaving diagnoses have been made. The nurse collaborates with the physician during TEE by ensuring adequate preparation, remaining at the patient's bedside to assist with and monitor the effects of probe insertion and ensuring close observation and postprocedural follow up. The complication rate for TEE is negligible. Possibilities include routine pediatric use, biplane imaging and incorporation with exercise stress testing. PMID- 8404006 TI - Drug-nutrient interactions in critically ill patients. PMID- 8404007 TI - Nursing Potpourri: one unit's way of sharing needed information. AB - The end product has been a more organized approach to communication with staff. Nursing Potpourri on Paper has replaced many notes, flyers and memos formerly posted on various boards and doors in the unit. Once the mechanism to generate it was in place, a small time commitment has been needed for production and distribution. Staff response has been positive, as evidenced by verbal comments and responses to offers in the newsletter. Some nurses save each issue. Many have contributed information or ideas. The newsletter has become a solution to some problems discussed in staff and management meetings. "We can put something about it in Nursing Potpourri" has been a frequent solution to communication needs. PMID- 8404008 TI - Updating the critical care nurse on alcohol and other drug abuse. PMID- 8404009 TI - Sickle cell disease: recent advances. PMID- 8404010 TI - Food allergies in children. AB - Ingested food represents the greatest foreign antigenic load that confronts the human immune system. In most individuals tolerance develops to food antigens that are continually gaining access to the body. When tolerance fails to develop, the immune system may react with a hypersensitivity reaction. Allergies to food affect up to 8% of children less than 3 years of age and 1% to 2% of the general population. Symptoms include the gastrointestinal, cutaneous, and respiratory symptoms, as well as systemic anaphylaxis with shock. Clinical investigations in the past have characterized the food hypersensitivity disorders, but our understanding of the basic immunopathologic mechanism remains incomplete. Current progress in allergen characterization and the rigorous scientific methods now being applied to this field by many investigators provide hope that new information regarding the pathogenesis of these disorders and new forms of therapy will soon become available. For now, practicing physicians must carefully diagnose specific food sensitivities and educate patients and their families in the elimination of the responsible food allergen. PMID- 8404011 TI - Acute rheumatic fever. PMID- 8404012 TI - Childhood asthma: diagnosis and treatment. AB - This discussion of asthma management should be regarded as providing guidelines, not dogma. The underlying principles of asthma management include recognition of the variability of the disease and the importance of the underlying inflammatory condition. Clinical assessment is not enough and objective monitoring with PEFR or spirometry provides important data. The treatment protocols require individualization. It is important that the patient and family are team members working together with the medical staff toward a goal of good asthma management. In the discussion of the management of asthma, much emphasis was placed on spirometry and home measurement of PEFR. Office use of spirometry is now the norm for asthma management. Providing asthmatic patients with peak flow meters and instructions in their use is part of the routine care of asthma. Instruction of the patient and family in the proper use of medications is paramount. The MDI devices need to be prescribed with careful instructions regarding their use. When the patient comes in for follow-up, part of the examination should include the patient's demonstration of how he uses this device. Discussion of the proper and safe use of bronchodilators is important. Overuse of inhaled bronchodilators may be a reflection of increasing asthma or, at the very least, evidence that the patient does not understand appropriate treatment of asthma. If a patient is dependent on regular use of an inhaled beta agent, it is likely that he would benefit from therapy directed at the underlying inflammation of asthma. The patient and the family should understand the purpose of each medication, the side effects, and the risks and benefits of their use. In particular, if steroid medications are necessary, the reasons for their use should be explained. Carefully matching the severity of the asthma with the therapeutic protocols provides an organized approach to asthma treatment. Avoiding triggers of asthma and controlling the environmental exposure to potential triggers leads to lower medication requirements and less lability. Offering the family written instructions to cope with changes in the child's condition, based on assessment of clinical and PEFR observations, allows them more autonomy and comfort in the day-to-day care of the asthmatic child. PMID- 8404013 TI - Malignant pleural mesothelioma. PMID- 8404014 TI - Recurrent pyogenic granulomas: a form of lobular capillary hemangioma. AB - Two cases of pyogenic granuloma recurred despite what was originally thought to be a sufficient surgical excision. The granulomas presented clinically as polypoid structures, one on the finger and one on the chin. On histologic examination, all showed polypoid configuration with lobules of small vessels and endothelial cell proliferation. The recurrent lesions also showed similar lobules, but interspersed between the collagen bundles in the deeper dermis. Vascular tumors such as the lobular pyogenic granulomas, intravenous capillary hemangiomas, and acquired tufted angiomas were compared to our tumors. PMID- 8404015 TI - Early skin metastasis of endometrial adenocarcinoma: case report and review of the literature. AB - A seventy-seven-year-old patient presented with skin metastasis from endometrial adenocarcinoma in a laparotomy scar, seven months after surgery. On reviewing current literature, it was clear that this was an unusual site for metastasis. We review the possible pathologic mechanism of this recurrence, its prognosis, and treatment. PMID- 8404016 TI - Dental curettes: useful tools for obtaining nail material for mycologic examination. AB - Obtaining material, especially subungually, from suspected onychomycotic nails can be difficult and/or traumatic with instrumentation frequently utilized by dermatologists. Use of scalpel blades and standard dermal curettes is often awkward and/or traumatic to the nail bed. Large clippings require micronization with a nail mill or tedious processing with less specialized instruments prior to microscopic examination (potassium hydroxide preparation) and fungal culture. A mycologist at our institution has efficaciously utilized dental curettes to obtain powdery subungual material suitable for immediate mycological assessment. This nontraumatic technique is becoming increasingly popular among clinicians. PMID- 8404017 TI - Kawasaki syndrome: a case report. AB - A four-year-old black boy with Kawasaki syndrome is reported. The child was treated with intravenous gamma globulin and aspirin. He had no disease-associated adverse sequelae. The clinical findings, diagnostic criteria, and treatment of Kawasaki syndrome are reviewed. PMID- 8404018 TI - Association of latex and food allergy. PMID- 8404019 TI - Sexually transmitted diseases: selections from the literature since 1990 syphilis: epidemiology. PMID- 8404020 TI - Childhood urticaria. PMID- 8404021 TI - A dermatologic diary. Portrait of a practice. PMID- 8404022 TI - Pericarditis associated with acral erythema of chemotherapy: a syndrome of cutaneous and serosal toxicities? AB - A nineteen-year-old woman whose Hodgkin's disease had relapsed experienced acral erythema in association with a asymptomatic pericardial friction rub following autologous bone marrow transplantation. An echocardiogram revealed a large pericardial and right pleural effusion. Since blood cultures gave negative results, renal function was normal, and the patient had neither neutropenia nor elevated temperature, an infectious cause was deemed unlikely and invasive procedures were not performed. These effusions resolved spontaneously. We propose that this patient's acral erythema and associated pericardial and pleural inflammation represent cutaneous and serosal toxic reactions to high-dosage chemotherapy that occur with the onset of leukocyte recovery. If so, acral erythema may signal the beginning of a toxic drug reaction. The appearance of erythema associated with lymphocyte recovery is due to immune hypersensitivity secondary to immaturity of the reconstituting immune system. Thus, we recommend that patients with acral erythema be examined for pleuropericarditis, especially if they experience chest pain. PMID- 8404023 TI - Skin manifestations of chronic obstructive pulmonary disease. AB - We report skin findings in thirty patients with severe chronic obstructive pulmonary disease. This study illustrates that patients with chronic obstructive pulmonary disease have numerous skin manifestations and some of these cutaneous findings are specific for chronic obstructive pulmonary disease. PMID- 8404024 TI - Generalized melanosis associated with malignant melanoma: unusual histologic appearance. AB - A case of generalized melanosis associated with malignant melanoma, characterized by up-to-date, previously undescribed histologic findings, is reported. Markedly dilated dermal lymphatics with features resembling secondary lymphangioma were found. We speculate that a further mechanism, as well as those previously reported in the literature, could be operative in the pathogenesis of this disorder of altered pigmentation. PMID- 8404025 TI - Zosteriform eruption of herpes simplex virus after vertebral metastases. AB - Described herein is a fifty-nine-year-old man with pulmonary adenocarcinoma metastatic to the spine who experienced a recurrent herpes simplex virus eruption in a zosteriform pattern on his lower back. This eruption corresponded to an area of the lumbosacral vertebrae that contained metastatic lesions. PMID- 8404026 TI - Measuring the repair of H2O2-induced DNA single strand breaks using the single cell gel assay. AB - The single cell gel (SCG) assay is for the detection of DNA single strand breaks and alkali labile sites in single cells. This investigation showed how the technique had an effective application in examining DNA single strand break repair. DNA from cultured human cells was damaged by treating the cells with H2O2, and samples were allowed to repair for various time intervals. The cells were then electrophoresed in a gel on a microscope slide, and the sizes of the individual DNA 'comets' were measured using laser scanning microscopy. Analysis of the comets indicated that actual DNA repair could be monitored over time. The SCG assay will help the study of DNA damage and repair, which will aid our understanding of the complex processes of mutagenesis and carcinogenesis. PMID- 8404027 TI - Response to thyroid stimulating hormone in 1-week-old rat thyroid gland after pretreatment with the same hormone in newborn conditions (hormonal imprinting). AB - Compared with control animals without hormone action, newborn rats treated with thyroid stimulating hormone (TSH) developed more exocytotic vesicles and enlarged endoplasmic reticulum filled with products in the thyroid gland up to the first postnatal week. A single administration of the hormone in newborn rats (imprinting) resulted in a long-lasting effect on the functioning of the cells of the thyroid gland. Single hormone action in postnatal 1-week-old animals provoked the discharge of products from the cells into the follicles of the thyroid gland with a concurrent endocytosis within 5 min after treatment. A similar but more vigorous effect was demonstrable in animals treated with TSH in newborn (hormonal imprinting) and postnatal 1-week-old conditions. Such events were accompanied by the death of certain cells while others developed myelin-like structures and showed signs of folliculogenesis in the cytoplasm. PMID- 8404028 TI - Chromosomal DNA extraction after restriction endonuclease treatments: a study by microdensitometry and electrophoresis. AB - The appearance of G- and C-banding patterns on metaphase chromosomes from the Don cell line (Cricetulus griseus) was accompanied by extraction of chromosomal DNA. A microdensitometric technique was employed to measure the loss of DNA produced by Hin dIII, Hae III and Eco RI endonuclease digestion on fixed chromosomes, from 0.5 to 24 h of treatment. Agarose gel electrophoresis was carried out to estimate the size of the DNA fragments extracted after digestion by Alu I and the restriction endonucleases already mentioned. Results obtained show that Alu I and Hae III, which possess 4 base pair recognition sequences, caused higher DNA extraction than enzymes with recognition targets of 6 base pairs (Hin dIII and Eco RI). Incubation buffers induced G-banding patterns which were accompanied by DNA extraction, although this loss was lower than that produced by endonuclease treatments. PMID- 8404029 TI - Polymorphonuclear cell-mediated phagocytosis and superoxide anion release in insulin-dependent diabetes mellitus. AB - It is well known that patients with insulin-dependent (or type I) diabetes mellitus are at high risk for bacterial infections. Since conflicting results have been reported on non-specific immune responses in type I diabetes, polymorphonuclear cell (PMN)-mediated phagocytosis and superoxide anion (O2-) generation in a group of individuals with well-controlled type I diabetes mellitus were assessed. Results showed that diabetic subjects were characterized by a significant impairment of phagocytic capacity when compared with healthy donors, while O2- release mimicked that seen in controls. Cell pretreatment with beta-hydroxybutyric acid (beta-HB) gave rise to a significant reduction in either phagocytosis or O2- production by PMN from type I diabetic individuals. Finally, beta-HB and glucose mixture supplementation to PMN suspensions did not induce any modification of their functional activities in comparison with those exerted by cells treated with beta-HB only. A disease-related or beta-HB-mediated PMN dysfunction in insulin-dependent diabetes mellitus is indicated. PMID- 8404030 TI - Thioacetamide-stimulated expression of non-histone protein. AB - The search for cancer specific nuclear proteins, stimulated by the supposition that transition from the normal to the neoplastic state resulting from disturbances in the control mechanisms of gene expression, indicated that non histone protein of MW 48 kD is much more abundant in animal tumour cells than in normal liver (Krajewska et al., 1990). A non-histone component of MW 48 kD was assessed for changes during chemically induced carcinogenesis. Rats were treated with the hepatocarcinogen thioacetamide (TAA) and the expression of the polypeptide studied, in total nuclear protein and nonhistone protein fractions, was tested by Western blot technique in the presence of antibodies developed against a component of MW 48 kD from Kirkman-Robbins hepatoma. It was demonstrated that TAA-induced hepatocarcinogenesis was accompanied by the expression of non-histone protein of MW 48 kD at a significantly elevated level. A clear and distinct change in the expression of the component studied in the spleen of TAA-treated rats was also observed. These results support the suggestion that over-expression of non-histone protein of MW 48 kD could contribute to neoplastic transformation. PMID- 8404031 TI - Origins of spontaneous mutants: cellular strategies for controlling their formation in response to changing environments. AB - Spontaneous and induced mutational events may not be as readily distinguishable as has been generally assumed. It is suggested that the later stages of the processes leading to the establishment of fixation of mutations may consist of a 'mutational pathway', with many features in common with the better-known metabolic pathways of organisms like Escherichia coli. If so, the mutational pathway may be controlled by catabolite repression, a cellular mechanism which switches metabolic pathways on and off (or more probably up and down) in response to intracellular levels of a molecular messenger known as cyclic adenosine monophosphate (cAMP). Mutation rates could then be regulated by an intracellular messenger in precisely the way which we would predict they ought to be regulated, if they are to cope with evolutionary pressures for increased variability. PMID- 8404032 TI - Thermodependence of human lymphocyte kinetics in vitro. AB - Lymphocytes from 35 healthy children were grown in cultures supplied with 5 bromodeoxyuridine at different temperatures (33-40 degrees C) for 48-120 h. The mitotic index and the distribution of the percentages of cells having passed one, two, and three or more DNA replications were quantified for each experimental condition. A replication index was calculated, whose dependence on temperature was depicted in the form of two Arrhenius plots (after 72 and 96 h of culture). Results showed the thermodependence of the lymphocyte kinetics. A break point at 36.1 degrees C was found in the two Arrhenius plots. The energies of activation (kJ/mol) under the break point, 197.2, and over the break point, 100.1 (72 h) and 43.1 (96 h), were calculated. PMID- 8404033 TI - Report and abstracts of the First International Workshop on Human Chromosome 8 Mapping. Vancouver, British Columbia, May 2-4, 1993. PMID- 8404034 TI - Report and abstracts of the Fourth International Workshop on Human X Chromosome Mapping 1993. St. Louis, Missouri, May 9-12, 1993. PMID- 8404035 TI - Mapping basigin (BSG), a member of the immunoglobulin superfamily, to 19p13.3. AB - Basigin is a novel member of the immunoglobulin superfamily ubiquitously expressed in various tissues. We mapped the basigin gene at 19p13.3, using a 1.6 kb cDNA fragment of the gene as a probe and sequential fluorescence in situ hybridization and G-banding on human metaphase chromosomes. PMID- 8404036 TI - Measuring genome reorganization from synteny data. AB - Two measures of the degree of genome reorganization based on synteny data are presented. The first, q, measures how far the original syntenic relationships have decayed during the separation of two taxa. The second measure, Q, is a logarithmic transformation of q, so constructed that Q increases unlimitedly while q deflects when reorganization becomes substantial. Numerical examples are given for the genomic reorganizations that have occurred during the evolution of humans, mice and rats. The degree of genome reorganization between the mouse and the rat is estimated to be about 1/5 of the degree of reorganization between humans and the rodent species. PMID- 8404037 TI - Mapping of the human adenylosuccinate lyase (ADSL) gene to chromosome 22q13.1- >q13.2. AB - Adenylosuccinate lyase (ADSL) is an essential enzyme involved in de novo purine biosynthesis. A deficiency in ADSL in humans has been shown to predispose to a neurodevelopmental syndrome with autistic features. Using both a somatic cell hybrid mapping panel and fluorescence in situ hybridization, we have precisely localized the human ADSL gene to chromosome 22q13.1-->q13.2. PMID- 8404038 TI - Assignment of the human endoglin gene (END) to 9q34-->qter. AB - Endoglin is a homodimeric membrane glycoprotein primarily associated with human vascular endothelium. It is also found on bone marrow proerythroblasts, activated monocytes and on lymphoblasts in childhood leukemia. Endoglin has recently been described as a component of the transforming growth factor-beta (TGF-beta) receptor system as it can bind TGF-beta 1 with high affinity. We now report on the localization of the human endoglin gene (END) to human chromosome 9, by Southern blot analysis of BglII fragments of DNA from human-hamster somatic cell hybrids. This chromosomal localization was confirmed by fluorescent in situ hybridization coupled with Distamicin A (DA)/4',6-diamidino-2-phenylindole (DAPI) banding on human chromosomes. The regional localization was assigned to 9q34- >qter by GTG-banding (G-banding by trypsin using Giemsa stain), indicating a telomeric position with respect to the Philadelphia breakpoint. PMID- 8404039 TI - Mapping of the bone morphogenetic protein 1 gene (BMP1) to 8p21: removal of BMP1 from candidacy for the bone disorder in Langer-Giedion syndrome. AB - Fifteen cosmids containing sequences from the human bone morphogenetic protein 1 gene (BMP1) were isolated from a cosmid library. The probe was a 483-bp DNA obtained by the reverse transcriptase-PCR method using primers designed according to the reported BMP1 sequence. When the positive cosmids were tested for chromosome fluorescence in situ hybridization, all showed signals at 8p21. The results indicated that BMP1 is not responsible for Langer-Giedion syndrome, whose putative gene has been assigned to 8q24. PMID- 8404040 TI - Localization of the transcription factor SP1 gene to human chromosome 12q12- >q13.2. AB - The cellular transcription factor SP1 binds to critical regulatory elements in a variety of cellular and viral promoters. The gene encoding the approximately 100 kDa SP1 protein contains zinc finger DNA-binding domains and glutamine-activation domains. Since SP1 is involved in the regulation of a variety of cellular genes, we wished to determine its chromosomal localization. Southern blot analysis of genomic DNA from a panel of mouse x human somatic cell hybrids indicated that SP1 was localized to chromosome 12. In situ hybridization allowed the localization of the gene encoding SP1 to human chromosome 12q12-->q13.2, with 12q13.1 being the most probable location. PMID- 8404041 TI - Detection of a male-specific sequence in nonhuman primates through use of the polymerase chain reaction. AB - Sex-specific DNA sequences are useful for detecting and monitoring chimerism in transplant recipients that had received sex-mismatched donor cells. Nonhuman primates are often used as experimental transplant models because of their evolutionary proximity, and the similarity of their physical characteristics, to those of humans. Unfortunately, DNA-based molecular detection strategies to monitor engraftment in sex-mismatched transplants in monkeys and baboons have not been available. We describe development of a polymerase chain reaction-based assay to detect a 174-bp male-specific sequence present in the rhesus monkey (Macaca mulatta) and olive baboon (Papio cynocephalus). The assay is sufficiently sensitive to allow detection of 10 male cells against a background of 10(4) female cells. Human sequence is not amplified under the described assay conditions. The amplified DNA sequence is 82% homologous to a sequence located near the testis-determining factor locus in the human genome, suggesting a high degree of evolutionary conservation in this region. PMID- 8404042 TI - Chromosomal localization of the Ox-44 (CD53) leukocyte antigen gene in man and rodents. AB - The Ox-44 (CD53) leukocyte antigen contributes to the transduction of CD2 generated signals in T cells and natural killer (NK) cells and has been suggested to play a role in growth regulation. The gene encoding this protein was assigned to the midportion of mouse chromosome 3 by interspecific backcross mapping and to human chromosome region 1p21-->p13.3 and rat chromosome region 2q34-->q41 by fluorescence in situ hybridization. Comparative mapping data presented in this report demonstrate conservation of synteny within the region encompassing this gene in mouse (Cd53), human (CD53), and rat (CD53). PMID- 8404043 TI - Isolation and characterization of somatic cell hybrids with breakpoints spanning 17q22-->q24. AB - Somatic cell hybrid mapping panels have previously been constructed to assist in the regional assignment of anonymous DNA probes and cloned genes to human chromosome 17. While a substantial number of hybrids are available that subdivide the short arm of this chromosome and the proximal portion of its long arm into specific regions, relatively few exist with breakpoints in the distal portion of the long arm. To increase the resolution of this region, four additional human x rodent somatic cell hybrids have been constructed that include breakpoints spanning the region 17q22-->q24. Hybrid clones carrying the long-arm derivative of chromosome 17 were initially identified by fluorescence in situ hybridization. Hybrids were subsequently screened using the polymerase chain reaction with primer sets representing DNA markers previously mapped to chromosome 17. These hybrids expand the existing somatic cell hybrid panel for the distal portion of the long arm of chromosome 17. PMID- 8404044 TI - Mapping the intron-containing human hsp90 alpha (HSPCAL4) gene to chromosome band 14q32. AB - The human heat-shock protein (hsp) 90 gene superfamily consists of two member forms, alpha and beta. We have mapped the intron-containing human hsp90 alpha expressed gene (HSPCAL4) to chromosome band 14q32. PMID- 8404045 TI - Localization of the gamma-subunit of the skeletal muscle L-type voltage-dependent calcium channel gene (CACNLG) to human chromosome band 17q24 by in situ hybridization and identification of a polymorphic repetitive DNA sequence at the gene locus. AB - The skeletal muscle dihydropyridine receptor consists of five subunits and fulfils an essential role in excitation-contraction coupling. A genomic clone for the human gamma subunit was used to map the gene (CACNLG) to chromosome band 17q24 by in situ hybridization. Contained within the gene is a 416-bp polymorphic repetitive DNA element that is potentially useful as a genetic marker. PMID- 8404046 TI - Localization of indoleamine 2,3-dioxygenase gene (INDO) to chromosome 8p12-->p11 by fluorescent in situ hybridization. AB - Indolamine 2,3-dioxygenase and tryptophan 2,3-dioxygenase are two enzymes that degrade tryptophan to N-formylkynurenine. The gene (TDO2) for tryptophane 2,3 dioxygenase has been localized to 4q31-->32. We now report localization of INDO (the gene encoding indolamine 2,3-dioxygenase) by fluorescent in situ hybridization to 8p12-->p11. PMID- 8404047 TI - Ordering of probes surrounding the Ewing's sarcoma breakpoint on chromosome 22 using fluorescent in situ hybridization to interphase nuclei. AB - Eight probes were localized by fluorescent in situ hybridization to the region surrounding the Ewing's sarcoma breakpoint on chromosome 22. Three of these were initially ordered by pair-wise hybridization to metaphase chromosomes with differential detection of the probes. These and the remaining probes were then ordered by hybridizing two or three probes simultaneously to interphase nuclei. In the two probe experiments and some of the three probe experiments, the order was derived by comparing mean interphase distances between signals from the probes. In the three probe experiments, either two probes were detected with one fluorochrome and the third with another or all three probes were individually distinguished by detecting one probe with fluorescein isothiocyanate (FITC), one with Texas Red, and one with both fluorochromes to give a mixed color. The order of the signals was then noted. Greater than 60 percent of configurations with a discrete order were shown or deduced to be correct. These approaches are assessed and we demonstrate a more obvious predominating order when all three probes are differentially detected. The order of the probes was deduced to be centromere: D22S271: D22S260: lambda S1: D22S262: cosK1831: Ewing's sarcoma breakpoint: cosLIF: D22S261: lambda S15: telomere. PMID- 8404048 TI - Tandem linkage of genes coding for leukemia inhibitory factor (LIF) and oncostatin M (OSM) on human chromosome 22. AB - Leukemia-inhibitory factor (LIF) and oncostatin M (OSM) are members of a family of structurally similar growth factors presenting overlapping and specific functions. Although the genes coding for IL-6, CSF3 and CNTF are scattered in the human and mouse genome, human LIF and OSM genes have conserved synteny in the course of evolution. Through isolation of a YAC and a cosmid clone containing both LIF and OSM we demonstrate that the two genes are linked in tandem on human chromosome 22q12, separated by 16 kilobases of intervening genomic DNA and transcribed in the same head-to-tail orientation. The close physical linkage between LIF and OSM genes brings new evidence of their evolutionary relationship. PMID- 8404049 TI - Fine mapping of the human receptor-like protein tyrosine phosphatase gene (PTPRM) to 18p11.2 by fluorescence in situ hybridization. AB - The isolation of a cDNA encoding a receptor-like protein tyrosine phosphatase, termed RPTP mu, and the assignment of the human gene to chromosome 18 has recently been reported (Gebbink et al., 1991). Using a genomic clone and fluorescence in situ hybridization, we now show that the human gene, designated PTPRM (formerly PTPRL1; LeBeau and Geurts van Kessel, 1991), maps in band 18p11.2. PMID- 8404050 TI - Genome distribution, chromosomal allocation, and organization of the major and minor satellite DNAs in 11 species and subspecies of the genus Mus. AB - We compared the genome distribution, chromosomal allocation, and organization of the major and minor satellite DNAs (satDNAs) in 11 species and subspecies of the genus Mus. Southern blot analysis of the major and minor satDNAs showed similar fragment profiles in all 11 species, with the exception of M. cervicolor and M. cookii for the major satDNAs and M. caroli, M. cervicolor, and M. cookii for the minor satDNAs, where these sequences could not be detected by the probes used. In situ hybridization of the major and minor satDNA probes revealed chromosome specific allocations of these sequences with quantitative species-specific patterns. Fluorometric analysis of the organization of the satellite sequences suggested that in the M. domesticus genome satDNA sequences are clustered in tandem repeats that are longer than those present in other Mus genomes. When compared with the other Mus genomes so far studied, the domesticus genome shows the highest quantity of satDNA sequences with a long-range organization of satDNA sequences. PMID- 8404051 TI - Karyotype of the giant mole-rat, Cryptomys mechowi (Rodentia, Bathyergidae). AB - The karyotype of the giant mole-rat, Cryptomys mechowi (Rodentia, Bathyergidae), from Zambia was investigated in one male and one female by means of G-, C-, and AgNOR-banding techniques. The diploid chromosomal set consisted of 40 biarmed chromosomes (2n = 40, NF = 80). A pair of autosomes in the male and the X chromosomes in the female were heteromorphic. The sex chromosomes were unusually large. PMID- 8404052 TI - Use of a subspecies cross for efficient development of a linkage map for a marsupial mammal, the tammar wallaby (Macropus eugenii). AB - The tammar wallaby (Macropus eugenii) has a 2n = 16 karyotype with an XX/XY female/male sex dimorphism. Female reproduction can be manipulated to produce up to five offspring per year. We have crossed two genetically distant subspecies of tammars, one from Kangaroo Island in South Australia, the other from Garden Island in Western Australia, to produce fertile F1 offspring of both sexes. Male F1 hybrids were crossed with female Kangaroo Island tammars to produce over 80 phase-known backcross progeny for linkage studies. Here we report detection of two linkage groups derived from these male recombination values. The first consists of alpha-lactalbumin and an anonymous tammar cDNA clone, pB72; the second contains the gene for a marsupial-specific milk protein (late lactation protein), the gene for lipoprotein lipase, and an anonymous cDNA clone, pB65. The gene for late lactation protein has previously been assigned to tammar chromosome 3. PMID- 8404053 TI - Recombination nodules and axial equalization in the ZW pairs of the Peking duck and the guinea fowl. AB - The meiotic Z and W chromosomes of the Guinea fowl (Numida meleagris) and the Peking duck (Anas platyrhynchos) show features similar to those described previously for 3 species of Phasianidae: chicken, Japanese quail and bobwhite quail. Axial equalization in the gonosomes of the Peking duck involves lengthening of the W axis along with shortening of the Z axis. A single recombination nodule is constantly found in the synaptic, terminal region of the ZW pairs stained with phosphotungstic acid. The localization of this nodule is significantly different in these two species, but the length of synaptonemal complex harboring the nodules is not significantly different. Based on the phylogenetic distance between the observed species, the present data support the hypothesis that the presence of a pseudoautosomal region is a general property of ZW pairs in carinate birds. PMID- 8404054 TI - Chromosomal anomalies and disturbance of transcriptional activity at the pachytene stage of meiosis: relationship to male sterility. AB - Morphological analysis of pachytene spermatocytes obtained from male mice carrying three chromosomal rearrangements--a Robertsonian translocation, Rb(X 2)2Ad; an autosomal reciprocal translocation, T(16;17)43H; and a tertiary trisomic, Ts(113)70H--demonstrated frequent association between the XY bivalent and the T43H and T70H translocation chromosomes. Quantitative autoradiographic data revealed that the normal transcriptional inactivity of the XY bivalent was not significantly disturbed, in contrast to that of the 16;17 quadrivalent and the extra 1(13) marker chromosome. These results are interpreted as an extension of the XY inactivation process to the associated autosomes and discussed in relation to male sterility. PMID- 8404055 TI - Localization of the alpha-S2-casein gene (CASAS2) to the homoeologous cattle, sheep, and goat chromosomes 4 by in situ hybridization. AB - The alpha-S2-casein gene (CASAS2) has been mapped to the homoeologous cattle, sheep, and goat chromosomes 4 and to the long arm of a bovine chromosome translocation, t(4;8), using nonradioactive in situ hybridization and simultaneous fluorescent R-banding. The t(4;8) has been characterized by GTG-, GBG-, and RBG-banding and by silver staining of nucleolus organizer regions. These results confirm the previous (ISCNDA, 1989) localization of the casein gene group to chromosome 4 of cattle, sheep, and goats. We propose that the discrepancy between our results and earlier assignments of the casein gene group to chromosome 6 of cattle and sheep can be explained by the fact that the chromosome identified as No. 6 in the Reading Conference (1976) report was renamed chromosome 4 in the ISCNDA (1989) standardized karyotype of both species. PMID- 8404056 TI - Flow cytometric detection of unbalanced ram spermatozoa from heterozygous 1;20 translocation carriers. AB - Flow cytometric analysis of enzymatically decondensed, DAPI-stained spermatozoa was performed to confirm the suspected production of unbalanced spermatozoa in heterozygous rams carrying a 1;20 translocation. High-precision flow cytometry (coefficient of variation, 0.6-0.8%) with a PAS II flow cytometer depicted Y- and X-chromosome-bearing spermatozoa from three cytogenetically normal rams as two distinct peaks. The difference in DNA fluorescence intensity between the gonosomes averaged 4.8%. Analysis of sperm samples from three heterozygous 1;20 translocation carriers yielded histograms with five peak distributions. The individual peaks were attributed to spermatozoa with a normal, balanced, and unbalanced chromosomal status. Peaks within Y- and X-spermatozoa populations were distributed in a ratio of 1:2:1 and were almost completely separated, with a coefficient of variation of 0.5-0.6%. Owing to the relative size of the translocated chromosomal segment (2.4% of the total DNA content, as determined from the flow cytometric data), histograms with five instead of the expected six peaks were observed. PMID- 8404057 TI - cDNA libraries from human tissues and cell lines. AB - We report the construction of 34 cDNA libraries from human tissues and cell lines in lambda phage vectors (Charon BS, lambda gt11, lambda SHK, or lambda zapII). The cDNA was synthesized from poly A+ RNA, using oligo-dT as a primer, and size selected before ligation to vector arms. High-complexity cDNA libraries from human tissues and cell lines should be a valuable resource for genome mapping studies and identification of disease genes. PMID- 8404058 TI - Assignment of the gene for human tear prealbumin (LCN1), a member of the lipocalin superfamily, to chromosome 8q24. AB - Tear prealbumin, a major protein of the human tear fluid, is also present in several other human body secretions. Recently it was shown to be a member of the lipocalin superfamily, a class of proteins that function as carriers for small hydrophobic molecules. Using a genomic tear prealbumin probe, we have mapped the gene (LCN1) to human chromosome 8q24 by fluorescent in situ hybridization. PMID- 8404059 TI - Assignments of 37 YAC clones to R-banded chromosomes by fluorescent in situ hybridization. AB - Fluorescent in situ hybridization was used to assign 37 yeast artificial chromosomes (YACs) from the CEPH (Centre d'Etude du Polymorphisme Humain) libraries to R-banded metaphases stained with propidium iodide. The use of the whole yeast DNA as probe, allows rapid mapping and characterization of YAC clones. Of 37 YAC clones, 11 were found cytogenetically chimeric (30%). The YAC clones reported in this study can be used to isolate and characterize targeted regions involved in genetic diseases and cancer breakpoints. PMID- 8404060 TI - Sequence-independent amplification and labeling of yeast artificial chromosomes for fluorescence in situ hybridization. AB - We have developed a method that allows reliable construction of high quality FISH probes from yeast artificial chromosomes (YACs) based on the separation of YACs by pulse-field gel electrophoresis and a rapid sequence-independent amplification procedure (SIA). These probes can be used to localize YACs on metaphase chromosomes and also with high efficiency, in interphase nuclei. PMID- 8404061 TI - Chromosome assignment of the human glutathione S-transferase mu 3 gene (GSTM3) to chromosome 1 by gene specific polymerase chain reaction. AB - Oligonucleotide primers specific for exons 4 and 5 sequences were used to amplify a unique 199-bp fragment in the human GSTM3 gene. Using DNA from a panel of somatic cell hybrids we assigned the GSTM3 locus to chromosome 1p. PMID- 8404062 TI - Isolation and mapping of 328 new cosmid markers on human chromosome 8: construction of a high-resolution cytogenetic map of chromosome 8 with 416 markers. AB - We have determined the chromosomal localizations of newly isolated cosmids by fluorescent in situ hybridization (FISH) on prometaphase R-banded chromosomes and have constructed a high-resolution cytogenetic map for human chromosome 8 with 416 cosmid markers, including 328 new markers and 88 reported previously. Of the 416 markers, 229 were mapped to the long arm of chromosome 8, 181 to the short arm, and 6 to the centromere. Although the clones were scattered throughout the chromosome, they were concentrated in R-positive bands. Since the estimated physical length of chromosome 8 is 135 Mb, the overall average distance between loci is 320 kb, but the average separation of loci on R-positive bands is nearly 130-200 kb. This cytogenetic map will serve as a resource for efforts to characterize chromosomal and molecular aberrations involved in cancers, to clone genes associated with hereditary diseases, and to construct a detailed physical map of large electrophoretic fragments and/or contiguous cosmids and yeast artificial chromosomes. PMID- 8404063 TI - Chromosome assignments of genes for rat ceruloplasmin (CP) and metallothionein (MT). AB - Chromosome assignments of the genes for rat ceruloplasmin (CP) and metallothionein (MT) were performed by analyzing somatic cell hybrid DNAs with the polymerase chain reaction (PCR), using primers specific for rat genes. The genes for CP and MT were assigned to rat chromosomes 2 and 19, respectively. PMID- 8404064 TI - Chromosome banding in amphibia. XX. DNA replication patterns in Gastrotheca riobambae (Anura, Hylidae). AB - Replication banding patterns were induced in the chromosomes of Gastrotheca riobambae by treating cell cultures with 5-bromodeoxyuridine (BrdU) and deoxythymidine (dT). In particular, the time sequence of replication of the highly differentiated XY/XX sex chromosomes was meticulously analyzed throughout the S phase. In the homogametic female, BrdU/dT labeling revealed no evidence of asynchronous replication between euchromatic regions in the XX pair. Minor replication asynchronies between the two X chromosomes were restricted to the heterochromatic C-bands. These results are further proof that dosage compensation in Amphibia for X- or Z-linked genes does not occur by inactivation of one of the two (XX or ZZ) sex chromosomes in the cells of homogametic individuals. In the very large and almost completely heterochromatic Y chromosome of G. riobambae, the various C-bands comprise at least three different structural categories, resulting in a complex pattern of early- and late-replicating bands along the Y chromosome. PMID- 8404065 TI - The RB gene locates on chromosome 15 at band 15q13 in the Syrian hamster. AB - The retinoblastoma susceptibility gene, which encodes a nuclear phosphoprotein (Rb), plays an important role in cell-cycle progression in both normal development and tumorigenesis. Using a Syrian hamster cDNA probe homologous to the human retinoblastoma gene (RB), we have localized the gene to chromosome band 15q13 by conventional in situ hybridization. PMID- 8404066 TI - High resolution ordering of DNA markers by multi-color fluorescent in situ hybridization of prophase chromosomes. AB - To improve resolution for physical ordering of adjacent DNA loci, prophase chromosomes were used for multi-color fluorescent in situ hybridization (FISH). The prophase chromosomes were prepared from cultured lymphocytes by a thymidine synchronization, bromodeoxyuridine release technique and then treating the synchronized cultures with topoisomerase II inhibitors ICRF154 or ICRF193. Almost all mitotic figures exhibited highly elongated prophase chromosomes without significant reduction of the mitotic index. Using multi-color FISH with these prophase chromosomes, we were able to distinguish signals for loci separated by as little as 50 kb, and determine their orientation. Furthermore, using this prophase ordering system, we confirmed the linear order and defined the orientation of seven cosmid markers within a 360-kb region surrounding D10S102, a locus that is closely linked to the disease locus in families segregating an allele causing multiple endocrine neoplasia IIA (MEN2A). This prophase FISH system, by rapidly and precisely providing the linear order of loci that are very close, can expedite construction of fine cytogenetic maps and contribute to positional-cloning studies in which the precise ordering of DNA loci in a target region is critical. PMID- 8404067 TI - Regional localization of 56 new human chromosome 18-specific yeast artificial chromosomes. AB - Fifty-six chromosome 18-specific yeast artificial chromosomes (YAC) were isolated from a human monochromosomal somatic cell hybrid DNA library. Fluorescence in situ hybridization revealed that the clones were evenly distributed throughout the chromosome. The clones, with an average insert size of 250 kb, cover about 18% of the euchromatic part of chromosome 18. Of 90 STS markers tested, 17 were represented in this YAC collection. Together with our previously reported set of unique chromosome 18-specific YACs, we now have available 111 regionally mapped, essentially nonchimeric, clones that provide more than 30% coverage and form a framework for the complete physical map. PMID- 8404068 TI - Localization of the highly polymorphic locus D19S120 to 19p13.3 by linkage. AB - To obtain the regional localization of a dinucleotide repeat D19S120 (formerly designated LIPE) with 7 alleles and an observed heterozygosity of 73% (Levitt et al., 1992) we typed 40 CEPH families. Given the high heterozygosity and ease of typing, this marker represents a useful addition to the index map of chromosome 19. PMID- 8404069 TI - Structure, expression and chromosome assignment of the human catenin (cadherin associated protein) alpha 1 gene (CTNNA1). AB - We have isolated the human alpha-catenin gene (CTNNA1), which encodes a cadherin associated protein, and have determined its primary structure and chromosomal localization. The transcript of CTNNA1 is 3.4 kb long and consists of 16 coding exons encoding 906 amino acids and at least one 5' noncoding exon. The 102-kDa predicted protein is the same size as the murine homolog, and the amino acid sequences of the two proteins are 99.2% homologous. Analysis by reverse transcription-PCR revealed that this gene is expressed ubiquitously in normal tissues. It was mapped to chromosome band 5q31 by fluorescent in situ hybridization. PMID- 8404070 TI - Induction of G-bands on Anguilla anguilla chromosomes by the restriction endonucleases HaeIII, HinfI, and MseI. AB - Fixed metaphase chromosomes of the eel Anguilla anguilla were treated with the restriction enzymes HaeIII, HinfI, and MseI. All three restriction endonucleases are capable of inducing a G-band-like pattern if the incubation time and enzyme concentration are controlled. The results are discussed in relation to the main factors involved in the appearance of serial G-bands. Chromatin organization, rather than DNA composition, seems to account for the interstitial G-band pattern exhibited by these eel chromosomes. PMID- 8404071 TI - Mapping of 18 probes on human chromosome 18 using single- and double-color FISH. AB - Fluorescence in situ hybridization (FISH) techniques were applied to the mapping of 18 probes from chromosome 18, permitting seven new assignments: D18S16 (18p11.31), D18S12 (18p11.1), D18S1 (18q12.2), D18S13 (18q21.31), D18S18 (18q21.31), D18S14 (18q21.33), and D18S17 (18q23). In addition, the localization of D18S3 in 18p11.3 was confirmed and that of D18S6, previously mapped in 18p11, was changed to 18q21.13. Finally, a more accurate mapping for nine probes was proposed: D18S7 (18q12.2), D18S10 (18q12.2), D18S24 (18q21.13), D18S8 (18q21.13), GRP (18q21.31), BCL2 (18q21.33), D18S5 (18q22.1), D18S19 (18q22.1), and D18S11 (18q22.3). The ordering of probes located in close proximity was made possible by combined use of single-color FISH with direct assignment on banded chromosomes, chromosomal length measurements, and double-color FISH. PMID- 8404072 TI - Chromosome assignment of human brain expressed sequence tags (ESTs) by analyzing fluorescently labeled PCR products from hybrid cell panels. AB - Sixty-three human brain cDNA sequences were newly assigned to individual human chromosomes. Ten of these were subregionally localized, and one was also mapped in the mouse genome. Four previously reported assignments were refined. PCR primers were designed from expressed sequence tags (ESTs) and tested for specific amplification from human genomic DNA. DNA was then amplified, often in multiplexed PCR reactions, using DNA from somatic cell hybrid mapping panels as templates. The amplification products were identified using an automated fluorescence detection system. Chromosomal assignments were made by discordancy analysis. Thirteen newly localized cDNAs exhibited homology to previously reported sequences. EST01471 was shown to correspond to human microtubule associated protein 1B (MAP1B), confirming the previous mapping of this gene to human chromosome 5. Other genes tentatively assigned to chromosomes based on these results were a component of the signal peptide receptor of the endoplasmic reticulum (EST00745) and a cyclic AMP-regulated phosphoprotein (EST01041) on chromosome 1, a protein phosphatase 2A 55-kDa regulatory subunit (EST01650) on chromosome 4, an NAD(P) transhydrogenase (EST01744) on chromosome 5, ribosomal proteins L1a or L1b (EST01627) and L18a (EST01583), a brain transcription factor (BF-1, EST00795) on chromosome 14, a milk fat globule membrane-related protein (EST01678) on chromosome 15, a putative peptide initiation factor (EST00675) on chromosome 17, thiosulfate sulfurtransferase (TST) on chromosome 22, and moesin (MSN) (EST00896) and a human equivalent of rat spot 14 (S14) (EST00887) on Xp11- >cen and Xpter-->p21.3, respectively. PMID- 8404073 TI - Detection of aneuploidy in human sperm by fluorescence in situ hybridization (FISH): different frequencies in fresh and stored sperm nuclei. AB - Fluorescence in situ hybridization (FISH) with chromosome-specific repetitive DNA probes provides a new method for rapid detection of aneuploidy in human sperm. There is widespread interest in this technique for basic research, as well as for screening men exposed to potential aneugens. A number of laboratories have reported a wide range in the frequency of aneuploidy for chromosomes in human sperm, suggesting that FISH may not reflect the true frequency accurately. We have serendipitously discovered that the length of time that fixed frozen sperm nuclei are stored affects both the hybridization efficiency and disomy frequency of individual chromosomes. This may explain some of the variation in the aneuploidy frequencies observed among laboratories. PMID- 8404074 TI - PCR probes for chromosome in situ hybridization of large-insert bacterial recombinants. AB - We have developed a procedure for efficient in situ hybridization of bacterial recombinants created with various types of large-insert cloning vectors. Minimal quantities of crude DNA are amplified and labeled during the degenerate oligonucleotide-primed polymerase chain reaction. The resulting probes generate high-intensity fluorescent hybridization signals on metaphase chromosomes and on interphase nuclei. PMID- 8404075 TI - BCG vaccination. Can it contribute to tuberculosis control? PMID- 8404076 TI - Progress in the control of hypertension. PMID- 8404077 TI - The Lung Health Study. Baseline characteristics of randomized participants. PMID- 8404078 TI - Don't drown the "down lung". PMID- 8404079 TI - Postpneumonectomy pulmonary edema. A retrospective analysis of associated variables. AB - STUDY OBJECTIVE: Evaluate the correlation between intravenous fluid administration and postpneumonectomy pulmonary edema. DESIGN: Retrospective chart review. SETTING: Large multispecialty group practice hospital. PATIENTS: Adults who had a pneumonectomy performed between 1977 and 1988. MEASUREMENTS AND RESULTS: Patients were identified who had postpneumonectomy pulmonary edema (PPE). Fluid administration and fluid balance information was found in records and compared with age- and sex-matched control patients who did not develop PPE. The side of pneumonectomy was noted for patients in each group. Autopsy findings were recorded for patients who died. Twenty-one patients met PPE criteria. No significant difference was found between groups for fluid administration or fluid balance. Patients who had right pneumonectomy had a significantly higher incidence of PPE. Patients with PPE had a 100 percent mortality rate and histologic evidence of the adult respiratory distress syndrome (ARDS) at autopsy. CONCLUSIONS: PPE is caused by noncardiogenic pulmonary edema rather than excess intravenous fluid administration. There is a greater incidence of the syndrome with right pneumonectomy for unknown reasons. The mortality rate is high despite interventions for ARDS. PMID- 8404080 TI - Extracorporeal life support for status asthmaticus. AB - Status asthmaticus is a life-threatening form of reactive airway disease, refractory to initial control with ordinary medical measures, sometimes requiring mechanical ventilatory assistance. In this report, we describe a patient whose bronchospasm could not be controlled with conventional measures, with severe respiratory acidosis (PaCO2 consistently > 100 mm Hg), who was successfully supported with extracorporeal life support (ECLS). During ECLS, arterial blood gas values rapidly returned to normal, and bronchospasm resolved during the subsequent 24 h. The patient was extubated hours later, and discharged to home 4 days later, neurologically normal. We believe that this represents the first application of this technology to this disease in an adult patient. PMID- 8404081 TI - A retrospective comparative study of in-hospital management of acute severe asthma: 1984 vs 1989. AB - Recent controversies examining the management of acute asthma prompted us to investigate whether there had been any significant changes in our management practices. We therefore audited the charts of all patients admitted to a large tertiary-care university-affiliated hospital with a primary diagnosis of acute asthma during the years of 1984 and 1989. A total of 67 patients' charts were reviewed (39 in 1984 and 28 in 1989). The mean age and initial flow rates (FEV1 or peak expiratory flow rate [PEFR]) were similar. In the emergency room, chest radiographs and arterial blood gas analyses were done more frequently than objective measures of flow. Fifty-one percent (20/39) of the patients had no measurement of flow in the emergency room in 1984 and 39 percent (11/28) in 1989 (p > 0.05). In both years, approximately 20 percent of the patients had no record of flow rates during their hospitalization (21 percent [8/39] in 1984 and 18 percent [5/28] in 1989). More studies of the blood were ordered in 1989, including hepatic enzyme and electrolyte measurements for no clear reasons. The clinical utility of chest radiographs was negligible. While the vast majority of patients received systemic corticosteroids in both years (85 percent [33/39] in 1984 and 96 percent [27/28] in 1989), 23 percent (9/39) and 18 percent (5/28) were discharged without oral steroid therapy in 1984 and 1989, respectively (p > 0.05). There was a significant decline in the use of aminophylline (95 percent [37/39] to 54 percent [15/28]; p < 0.05) and an increase in the use of ipratropium bromide (15 percent [6/39] to 75 percent [21/28]; p < 0.05) in 1989. Theophylline levels were less likely to be measured in 1989, and the majority of levels in both years were either subtherapeutic or toxic. No patients were discharged with peak flow meters or recorded action plans, although follow-up arrangements were recorded in 87 percent (34/39) and 96 percent (27/28) of the patients in 1984 and 1989. We conclude that while improvements in in-hospital management of asthma were noted in 1989, suboptimal management practices are still common. PMID- 8404082 TI - Difficult-to-control asthma. Contributing factors and outcome of a systematic management protocol. AB - To our knowledge, there are no published results of protocols for managing difficult to control asthma (DTCA) or of the spectrum and frequencies of reasons why asthma can be difficult to control (DTC). To assess the usefulness of a systematic management protocol and determine the reason(s) why asthmatics are DTC, we developed a protocol that systematically considered multiple factors that may make asthma worse and prospectively evaluated the outcomes of therapeutic interventions for these factors and of inhaled corticosteroids (ICS) and azathioprine. We studied 42 consecutive and unselected DTCAs (19 men and 23 women) whose age was 48 +/- 15.9 years. They had a diagnosis of asthma for 15.1 +/- 15.8 years, were DTC for 4.8 +/- 7.8 years, and were followed up by us in the study for a total of 3.5 +/- 1.9 years. Initially, the dose of prednisone was 30.2 +/- 22 mg/d. Following utilization of the protocol, 74 percent were no longer DTC. It took 1.8 +/- 1.7 years for them to no longer be DTC; they remained so for 1.8 +/- 1.5 years. In these patients, 2.7 +/- 1.2 factors appeared to be responsible for the DTC state; 80 percent had > or = 2. Improvement was more likely if gastroesophageal reflux (GER) was a factor (p = 0.014); it correlated with the addition of ICS (p = 0.04) and treatment for GER (p = 0.02). Failure to reverse DTCA correlated with the suspicion (p = 0.004) and admission of nonadherence (p = 0.04). In 14 patients given azathioprine, prednisone dose decreased from 45 +/- 25.3 to 13.3 +/- 21.6 mg/d (p = 0.003); 6 of 14 achieved no longer DTC status; and substantial morbidity occurred. The reason(s) for DTCA could be determined in most instances by utilizing a systematic protocol; multiple factors were responsible in the majority of cases; treatment for GER and ICS were the two most helpful interventions; nonadherence was the most likely reason suspected for maintaining DTCA; and azathioprine acted as a corticosteroid sparing agent that should not be prescribed routinely. PMID- 8404083 TI - Results of screening for tuberculosis in foreign-born persons applying for adjustment of immigration status. AB - As part of the required screening process of illegal aliens applying for adjustment of status by the Immigration and Naturalization Service, 7,573 persons were evaluated for tuberculous infection by the Denver Department of Health and Hospitals from May 1987 through December 1988. Applicants were screened with tuberculin skin testing, chest radiographs, or both. Review of 6,520 charts that were available found that 4,840 applicants had tuberculin skin tests, of which 2,039 (42 percent) were > or = 10 mm and 1,528 had further evaluation at the Denver Metro Tuberculosis Clinic. Seventy-five percent of the applicants were between the ages of 15 and 34 years, and 91 percent were from Mexico. Evidence of past or current tuberculous infection on chest radiograph was present in 273 (17 percent) and 16 (7 percent) had sputum cultures obtained that identified four new cases of active tuberculosis. Isoniazid preventive therapy (IPT) was recommended to 1,029 applicants, of whom 29 (3 percent) were 35 years of age or older; 716 (70 percent) completed at least six months of treatment. We conclude that there is a high prevalence of tuberculous infection in foreign-born persons applying for adjustment of immigration status, but a low prevalence of clinically apparent tuberculosis. This population is an excellent target for IPT, which can be achieved with good success. Proactive screening and preventive therapy is likely to significantly reduce tuberculosis reactivation and morbidity, prevent secondary infection of contacts, and be cost-effective. PMID- 8404084 TI - Self-reported vs measured compliance with nasal CPAP for obstructive sleep apnea. AB - To estimate reliability of self-reported compliance with nasal continuous positive airway pressure (CPAP) therapy for obstructive sleep apnea (OSA), we studied 63 OSA patients aged 53.7 +/- 1.2 years (mean +/- SEM) with an apnea hypopnea index (AHI) of 50.8 +/- 2.9 and lowest sleep SaO2 of 65.6 +/- 2.3 percent receiving nasal CPAP for 539 +/- 44 days. During a follow-up polysomnography (PSG) on the pressure prescribed for home therapy (10.3 +/- 0.3 cm H2O), the hours of operation shown on the built-in time counter of the patients' devices were read to determine objective compliance by dividing the run time by the days since initiation of therapy. This parameter was compared with subjective compliance reported in a self-administered questionnaire. Mean measured use time was 4.9 +/- 0.3 h per night, whereas reported daily use time calculated from reported nights a week and hours a night was 6.1 +/- 0.3 h per night. As predominantly patients with poor compliance misestimated daily use time, we conclude that self-reports are unable to distinguish between compliant and noncompliant patients. PMID- 8404085 TI - Pulmonary edema after anteroseptal acute myocardial infarction. AB - To evaluate the clinical characteristics of patients with anteroseptal myocardial infarction (MI) initially presenting with pulmonary edema, we analyzed 58 patients with anteroseptal MI who underwent emergency coronary arteriography that revealed single-vessel disease of the left anterior descending coronary artery. Of the 58 patients, pulmonary edema was observed in 24 patients (group A) and was absent in 34 patients (group B). Pulmonary capillary wedge pressure was significantly higher, and cardiac output was significantly lower in group A. The site of coronary stenosis, maximum serum creatinine kinase value, and wall motion point score did not differ between the two groups. However, the incidence of previous hypertension and posterior wall thickness > or = 11 mm was significantly higher in group A than in group B (p < 0.001 and p < 0.05, respectively). Thus, impaired left ventricular diastolic filling in the non-MI segments due to higher incidence of hypertension and left ventricular hypertrophy was considered to be the possible cause of pulmonary edema. PMID- 8404086 TI - Blood gas dynamics at the onset of exercise in heart transplant recipients. AB - One hypothesis to explain the rapid neural component of exercise hyperpnea contends that afferent stimuli originating in the ventricles of the heart act reflexly on the respiratory center at the onset of exercise, ie, "cardiodynamic hyperpnea." Orthotopic cardiac transplantation (Tx) results in the loss of afferent information from the ventricles. Thus, Tx possibly results in transient hypercapnia and hypoxemia in deafferented heart transplant recipients (HTR) at the onset of exercise due to hypoventilation. To examine the cardiodynamic hypothesis, we collected serial arterial blood gas (ABG) samples during both the transient and the steady-state responses to moderate cycle exercise in 5 HTRs (55 +/- 7 years) 14 +/- 7 months post-Tx and 5 control subjects matched with respect to gender, age, and body composition. Forced vital capacity, forced expiratory volume in 1 s, total lung capacity, and diffusion capacity did not differ (p > or = 0.05) between groups. Resting arterial PO2, PCO2, and pH did not differ between groups (p > or = 0.05). The ABGs were drawn every 30 s during the first 5 min and at 6, 8, and 10 min of constant load square wave cycle exercise at 40 percent of the peak power output (watts). Absolute and relative changes in arterial PO2, PCO2, and pH were similar (p > or = 0.05) between HTR and the control group at all measurement periods during exercise. Heart rate (%HRmax reserve), rating of perceived exertion, and reductions in plasma volume (% delta from baseline) did not differ between HTR and control during exercise at 40 percent of peak power output (p > or = 0.05). Our results demonstrate that there is no discernible abnormality in ABG dynamics during the transient response to exercise at 40 percent of peak power output in patients with known cardiac denervation. These data do not support the cardiodynamic hyperpnea hypothesis of ventilatory control in humans. The absence of hypercapnia in HTRs is further evidence for the existence of redundant mechanisms capable of stimulating exercise hyperpnea. PMID- 8404087 TI - Symptoms, lung function, and airway responsiveness following irritant inhalation. AB - STUDY OBJECTIVE: To estimate the incidence of respiratory symptoms and physiologic abnormalities after inhalation of common irritant chemicals. DESIGN: Structured interview of 12 months of poison control center (PCC) inhalation cases. Follow-up measurement of pulmonary function and airway responsiveness in a subgroup with symptoms 12 to 24 h postexposure. SETTING: A regional PCC and clinical pulmonary function laboratory. PATIENTS: Consecutive sample of 547 inhalation cases. Interviews of 299 subjects. Lung function follow-up in 10 subjects. MEASUREMENTS AND MAIN RESULTS: Immediate respiratory symptoms were reported by 262 (88 percent) subjects; 12 to 24 h postexposure symptoms were reported by 130 (44 percent). Cigarette smoking was significantly related to immediate onset of cough (relative risk [RR] = 1.3; 95 percent confidence interval [CI], 1.1 to 1.5); both smoking (RR = 1.6; 95 percent CI, 1.1 to 2.1) and prior asthma (RR = 1.3; 95 percent CI, 1.1 to 1.6) were associated with wheeze, exhibiting multiplicative combined risk (RR = 2.8; 95 percent CI, 1.9 to 4.3). Of 10 subjects studied, none had abnormal airflow or lung volumes 8 +/- 4 days postexposure; 8 demonstrated increased airway responsiveness to methacholine. By three months, only one subject's increased responsiveness reversed; in three others, symptoms resolved but increased responsiveness remained. CONCLUSIONS: Respiratory symptoms following irritant exposure are associated with smoking and asthma and typically resolve quickly. Continuing symptoms are associated with persistent increased airway responsiveness without other pulmonary function abnormalities. This may reflect newly induced airway changes or, alternatively, could represent underlying increased responsiveness in subjects symptomatic after irritant exposure. PMID- 8404088 TI - Heart transplantation after cardioverter-defibrillator implantation. A case control study. AB - A case control study was performed to determine whether previous implantable cardioverter-defibrillator (ICD) insertion adversely affects outcome after heart transplantation. Six male heart transplant recipients who had undergone ICD insertion 12 +/- 5 months before heart transplantation were compared to a cohort of six heart transplant recipients who were matched according to age, preoperative status and hemodynamics, date of transplantation, graft ischemic time, history of a previous cardiac operation, and duration of follow-up. There were no significant differences in operating room time, chest tube drainage, time to extubation, and the duration of intensive care unit or hospital stay between the two groups. Furthermore, there were no significant differences in the number of units of packed cells, fresh frozen plasma, platelets and cryoprecipitate transfused. The number of treated rejection episodes and the number of patients requiring intravenous antibiotics for infection in the first 90 days was identical between groups. It was concluded that heart transplantation after ICD implantation did not appear to carry more risk than heart transplantation after a previous cardiac operation. Our limited experience supports the potential use of the ICD in patients with life-threatening ventricular dysrhythmias who are awaiting transplantation. PMID- 8404089 TI - Progression of aortic stenosis. Role of age and concomitant coronary artery disease. AB - To determine the spontaneous progression of valvular aortic stenosis and to delineate clinical factors related to progression, a longitudinal study, including 49 patients (aged 16 to 81 years), was performed. All patients had auscultatory findings of aortic stenosis and multiple Doppler echocardiograms separated by at least 11 months. Rapid progression of aortic stenosis was defined as an increase of maximal instantaneous pressure gradient by > or = 10 mm Hg per year. During a mean follow-up period of 32 months (11 to 66 months), maximal pressure gradient rose from 38 +/- 15 to 60 +/- 20 mm Hg in the entire study population, resulting in a median increase of 7.2 mm Hg per year. In 21 patients (43 percent), an increase of > or = 10 mm Hg per year was found; in this subgroup with rapid progression, patients were older (64 vs 53 years, p < 0.01) and coronary artery disease was more prevalent (38 percent vs 7 percent, p = 0.01). We conclude that nearly half the patients with initially mild to moderate valvular aortic stenosis reveal a progression of > or = 10 mm Hg per year. PMID- 8404090 TI - Differential diagnostic value of plasma cells in bronchoalveolar lavage fluid. AB - The aim of this study was to investigate whether the demonstration of plasma cells (PC), which are normally absent in bronchoalveolar lavage (BAL) fluid, facilitates differentiation among pulmonary disorders. Initial BAL fluid samples of 1,260 patients were analyzed. In 83 of these, PC were found. Of these 83, 47 were obtained from individuals suffering from extrinsic allergic alveolitis (EAA). The number of PC in BAL fluid from EAA patients was found related to the time between antigen exposure and BAL. Drug-induced pneumonitis appeared to be another disorder with a high percentage of cases with PC in the BAL fluid (35.7 percent). Therefore, we conclude that determination of PC in BAL fluid has differential diagnostic value in discriminating among interstitial lung diseases of various origins. However, the exact role of BAL fluid and PC and the link to clinical manifestations of these diseases needs further investigation. PMID- 8404091 TI - Dose dependency of aminophylline effects on hemodynamic and ventricular function in patients with chronic obstructive pulmonary disease. AB - The effects of aminophylline on pulmonary vascular tone, systemic hemodynamics, and ventricular ejection fractions reported in the literature show some discrepancies. We therefore studied in COPD patients the effects of aminophylline on hemodynamics, on ventricular ejection fractions, and on systolic and diastolic functions of each ventricle, and we measured simultaneously the blood level of the drug. The analysis of the data revealed a relationship between the blood level of aminophylline and the variations of right ventricular ejection fraction (RVEF) (r = 0.83, p = 0.005), left ventricular ejection fraction (LVEF) (r = 0.76, p = 0.017), pulmonary vascular resistance index (PVRI) (r = -0.58, p = 0.096), systemic vascular resistance index (SVRI) (r = -0.60, p = 0.08), and right ventricular peak systolic pressure/end-systolic volume index (RVPSP/ESVI) (r = -0.75, p = 0.02). Modifications of ejection fractions and vascular resistance indices were correlated for both ventricles (RVEF vs PVRI, r = -0.77, p = 0.01; LVEF vs SVRI, r = -0.76, p = 0.02). Finally, RVEF modifications was also correlated to RVPSP/ESVI variation (r = 0.78, p = 0.01). These results suggest that even within the therapeutic range (10 to 20 mg/L), the effects of aminophylline seemed to depend on its blood level. This dose dependency could explain the contradictory data reported in the literature concerning the effects of aminophylline on pulmonary and systemic hemodynamics and on ventricular function. PMID- 8404092 TI - Phenotypes and lymphokine-activated killer activity of pleural cavity lymphocytes of lung cancer patients without malignant effusion. AB - We examined the phenotypes of lymphocytes in the pleural cavity of 23 lung cancer patients without malignant effusion. The ability of those lymphocytes to develop lymphokine-activated killer (LAK) activity and the regulation of LAK by pleural cavity macrophages were also compared with their counterparts in the peripheral blood. Mononuclear cells (MNC) were obtained simultaneously from the blood and by lavage of the pleural cavity of patients with lung cancer. The proportion of the T-cell subset of HLA-DR+ cells was significantly higher in the pleural cavity than in the peripheral blood, but the proportions of CD3+ and CD8+ cells in the pleural cavity were similar to the corresponding proportions in the blood. The proportions of CD4+ and CD16+ cells were lower in the pleural cavity than in the blood. The LAK activity could be developed by MNC from the pleural cavity following incubation with interleukin 2 (IL-2), but the LAK activity of pleural cavity MNC was significantly less than that of peripheral MNC. Pleural cavity lymphocytes alone also developed LAK activity following incubation with IL-2. Pleural macrophages from the patients were regulated to augment in vitro induction of LAK activity by IL-2 from autologous blood lymphocytes and pleural cavity lymphocytes. Lymphocytes in the pleural cavity without malignant pleural effusion could be developed by LAK activity and this activity was augmented by pleural cavity macrophages. The LAK activity developed by pleural cavity lymphocytes was significantly lower than that developed by peripheral blood lymphocytes. However, they can change their population to include cells with higher activities on exposure to IL-2 against the invasion of lung cancer cells into the pleural cavity. Thus, the population of lymphocytes in the pleural cavity of patients with lung cancer without malignant pleural effusion was different from that in malignant pleural effusion. PMID- 8404093 TI - Terminal pulmonary infections in patients with lung cancer. AB - To determine the factors that predispose the patient with lung cancer to develop terminal pulmonary infections, we reviewed the case records and autopsy data of 304 patients who died of lung cancer in the Kyushu University Hospital between 1976 and 1990. The incidence of mycobacterial infection was significantly higher among those treated with antineoplastic therapy and corticosteroids (group 3) than in those who received antineoplastic therapy alone (group 2). The incidence of nonbacterial infection did not differ significantly between the two groups. In some group 3 patients, the administration of corticosteroids for relatively short periods (less than one month) led to fatal mycobacterial infection. Among those patients with lymphocytopenia, the incidence of fatal mycobacterial infection was significantly higher in group 3 than in group 2, whereas the incidence of fatal nonbacterial infection was not. In group 3, the incidence of fatal mycobacterial and nonbacterial infections did not differ significantly among those with and without lymphocytopenia. Thus, in patients with lung cancer who were receiving antineoplastic treatment, corticosteroids were more closely associated with the development and exacerbation of mycobacterial infection than was lymphocytopenia. The influence of corticosteroids on the development of nonbacterial infection was not more marked than that of lymphocytopenia. The incidence of common bacterial infections was no higher among those patients who received no antineoplastic treatment or corticosteroid (group 1), group 2, and group 3. Therefore, the local and systemic effects of the lung cancer itself are likely more important in predisposing the patient to bacterial infections than are either antineoplastic agents or corticosteroids. PMID- 8404094 TI - Bronchoscopic diagnosis of pulmonary infections in a heterogeneous, nonselected group of patients. AB - Fiberoptic bronchoscopy with bronchoalveolar lavage and protected specimen brush technique has become an established method for etiologic diagnosis in severe forms of pulmonary infections during recent years. In this study, including 62 bronchoscopies in 53 patients, a standardized program, covering all important pulmonary pathogens, has been evaluated in a heterogeneous group of patients. Results providing therapeutic guidelines were obtained in 53 percent (16/30) of the immunocompromised patients (including 5 bronchoscopies on HIV-positive patients), but only 19 percent (6/32) of the immunocompetent patients (p < 0.001). We conclude that bronchoscopy is of great value for diagnosing pulmonary infections in immunocompromised patients. In immunocompetent patients, the diagnostic yield is lower and the indication for bronchoscopy must be established for each individual patient based on clinical importance, resources, and risk. When bronchoscopy is performed, we believe that a standardized program like ours reduces the risk of missing important pathogens. PMID- 8404095 TI - Effect of infarct site on diastolic time during exercise. AB - To assess the difference in left ventricular performance during exercise between anterior (11 patients) and inferior (10 patients) myocardial infarction (MI) of equivalent size, patients performed a supine bicycle exercise 6 to 8 weeks after the first acute MI. All patients had negative exercise test results and despite no significant differences in HR, blood pressure and stroke volume index at peak exercise, pulmonary artery wedge pressure was significantly higher in anterior (35 +/- 7 mm Hg) than in inferior MI (27 +/- 9 mm Hg). Although there were no significant differences in electromechanical systole (QS2) and diastolic time (DT) at rest, a significant prolongation of QS2 with consequent shortening of DT (p < 0.01) was observed at peak exercise in anterior MI. In addition to decreased subendocardial coronary blood flow from increased left ventricular end-diastolic pressure, a disproportionate shortening of DT in anterior MI may initiate subendocardial ischemia in the noninfarcted segments, which may further impede subendocardial blood flow. PMID- 8404096 TI - The effect of pyridostigmine on bronchial hyperreactivity. AB - We examined the effect of pyridostigmine (PY) at a dose of 30 mg orally three times a day on nonspecific bronchial hyperreactivity in ten normal nonsmokers (NNS), ten smokers (SM), and ten mild asthmatics (AS). We conducted a double blind, placebo-controlled, crossover trial, randomly assigning subjects to receive either placebo (PL) or PY before undergoing bronchoprovocation challenge with eucapnic voluntary hyperventilation (EVH) using dry gas. Compliance with PY was confirmed by measuring red blood cell acetylcholinesterase (Achase) levels during both days of testing. While taking PL, the mean (+/- SEM) falls in FVC and FEV1 after the bronchoprovocation were as follows: NNS, 1.0 percent (+/- 0.6) FVC and 4.3 percent (+/- 1.0) FEV1; SM, 2.4 percent (+/- 1.1) FVC and 2.7 percent (+/ 1.3) FEV1; AS, 5.3 percent (+/- 2.3) FVC and 11.5 percent (+/- 2.8) FEV1. The mean decreases in FVC and FEV1 while taking PY were as follows: NNS, 1.8 percent (+/- 0.7) FVC and 4.3 percent (+/- 0.8) FEV1; SM, 3.8 percent (+/- 1.4) FVC and 5.2 percent (+/- 1.6) FEV1; AS, 4.4 percent (+/- 1.3) FVC and 11.8 percent (+/- 2.8) FEV1. Within each category, using a paired t test to compare the results on each day of testing, no statistically significant differences were noted. Pyridostigmine at the tested dose has no significant effect on nonspecific bronchial hyperreactivity in normal NNS, SM, or AS. PMID- 8404097 TI - Round atelectasis and Metsovo lung. AB - Round (helical) atelectasis is one of the benign sequelae of occupational asbestos exposure. Environmental asbestos exposure does not differ from occupational in its pleural manifestations, but to our knowledge, round atelectasis has not been reported yet. In the present study, we present the clinical and radiologic findings of five individuals with round atelectasis. They were all born in the Metsovo area, northwest Greece, where environmental exposure to asbestos (tremolite) has been documented. All five had negative evaluation for malignancy. In addition, they have been followed up for one to four years and four of them are in good health, thus confirming round atelectasis as a benign, nonpremalignant condition. The fifth patient died of malignant pleural mesothelioma two years later, while the previously detected round atelectasis remained unchanged. We therefore consider that his mesothelioma was not related to the round atelectasis, although both were certainly related to the same environmental asbestos exposure. PMID- 8404098 TI - Augmented resting sympathetic activity in awake patients with obstructive sleep apnea. AB - Muscle nerve sympathetic activity (MSA) was recorded during wakefulness in 11 patients with obstructive sleep apnea (OSA) and in 9 sex- and age-matched healthy control subjects. Plasma levels of norepinephrine (NE) and neuropeptide Y were analyzed. Five patients had established hypertension (resting supine systolic BP/diastolic BP > or = 160/95 mm Hg). The investigation was performed after a minimum of 3 weeks' washout period of antihypertensive medication. Muscle sympathetic activity during supine rest was higher in patients compared with controls (p < 0.01) with no difference between normotensive and hypertensive patients. However, systolic, but not diastolic, BP was positively related to resting MSA (n = 20, p < 0.01). There was no significant correlation between body mass index and MSA. Resting MSA was unrelated to disease severity expressed as apnea frequency or minimum SaO2 during the overnight recording. Both the arterial and venous plasma norepinephrine was higher in patients compared with controls (p < 0.05). Plasma levels of NE correlated to resting MSA (p < 0.01) in the whole study group (patients and controls) but not within the respective subgroups. No significant correlation, however, was found between plasma NE (arterial and venous) and BP. Plasma neuropeptide Y-like immunoreactivity was similar in patients and controls. However, one patient with hypertension had approximately twice this level in repeated samples. It is concluded that neurogenic sympathetic activity as well as circulating plasma NE is increased in patients with OSA. This increased sympathetic activity during awake supine rest may reflect a pathophysiologic adaptation to hypoxia and hemodynamic changes occurring at repetitive apneas during sleep. The correlation between MSA and systolic BP implies that this mechanism may be directly or indirectly involved in the development of cardiovascular complications in OSA. PMID- 8404099 TI - Comparison of snoring measured at home and during polysomnographic studies. AB - Snoring characteristics depend on several factors (sleep position, sleep architecture, breathing route) that can be influenced by changes in sleep habits and by the presence of the different probes and electrodes during polysomnographic studies. Our objective in this study was to compare the characteristics of snoring in the home environment with those of the sleep laboratory where most conventional studies are carried out. Fourteen nonapneic snorers were subjected to three night recording sessions within a two-week period, two at home and one in the sleep laboratory. To eliminate any sleep interference by the apparatus, breathing sounds were recorded with two microphones symmetrically placed on either side of the bed, the signal being preamplified and stored on a VHS hi-fi video recorder. The recorded signal was analyzed by using a spectrum analyzer (real time analyzer) and an equalizer to correct for acoustic resonances of the bedrooms. A snoring event was defined as a breathing sound with a sound pressure level (SPL) greater than 60 dB SPL. The snoring index (number/sleep hour) and the sound intensity of each event were automatically determined. The total sleep time (TST) was similar for the two home recordings (6.8 +/- 0.2 and 7.0 +/- 0.2 h, respectively, mean +/- SEM), but it was significantly shorter during the hospital study (6.0 +/- 0.3 h). The snoring indices obtained at home were 141.4 +/- 33.3 and 144.1 +/- 41.2/h and not statistically different from those obtained during the hospital recording (209.1 +/- 41.5/h). The percentage of TST spent above 60 dB SPL was significantly greater during the polysomnographic study (4.3 +/- 1.2 percent) than during the home recordings (2.5 +/- 0.7 and 2.9 +/- 1.0 percent, respectively). We conclude that the severity of snoring may be overestimated during polysomnographic recordings. PMID- 8404100 TI - Effects of inhaled anticholinergic drug on dyspnea and gas exchange during exercise in patients with chronic obstructive pulmonary disease. AB - To elucidate the effect of oxitropium bromide (OTB), an anticholinergic drug, on dyspnea and gas exchange during exercise in patients with chronic obstructive pulmonary disease (COPD), we performed the cycle exercise test on 19 patients with COPD (mean age, 72.0 +/- 1.9 years; mean FEV1, 1.28 +/- 0.07 L) before and after inhalation of OTB, 300 micrograms, or placebo, 300 micrograms, in randomized fashion. Spirometry was performed immediately before and 30 min after inhalation of either OTB or placebo. Dyspnea during exercise was evaluated using the Borg scale (BS) and the slope of the regression between BS and oxygen uptake (VO2) during exercise (Borg scale slope: BSS). Arterial oxygen saturation (SaO2) was continuously monitored by pulse oximeter during and after exercise. We also measured the recovery time, which was defined as the time to recover decreases in SaO2 after exercise. After OTB, spirometric indices were improved (delta FEV1 16.8 +/- 0.9 percent) and maximal VO2 during exercise increased significantly (from 986 +/- 46 ml/min to 1,156 +/- 55 ml/min, p < 0.01), but not after placebo. The maximal scores of BS and the BSS were significantly decreased after OTB, but not after placebo. Although the SaO2 at rest and during exercise did not differ with or without either OTB or placebo, the recovery time after OTB (77.3 +/- 6.8 s) was significantly shorter than that before administration (98.4 +/- 14.6 s) (p < 0.01). We conclude that the inhaled OTB produces small but significant improvement both in dyspnea during exercise and in exercise performance in stable COPD and may contribute to improve the quality of life in some patients with COPD. However, gas exchange during exercise of COPD patients is little affected by OTB. PMID- 8404101 TI - Local airways immune modifications induced by oral bacterial extracts in chronic bronchitis. AB - Bacterial extracts can act as immune stimulants and in some instances have been used, rather empirically, to prevent recurrent infections in the nonimmunocompromised host. Some agents are administered via oral route with the goal to increase airways immune defenses. In animal models and in normal humans, gut-associated lymphoid tissue (GALT) stimulation is able to induce a generalized response by the whole mucosal-associated lymphoid tissue (MALT). The aim of this placebo-controlled, double-blind, parallel-group study was to evaluate whether the stimulation of the GALT through oral administration of a polyvalent bacterial extract (BE) could lead to significant immune modifications either systemically or locally in the respiratory tract in patients suffering from chronic bronchitis. We selected 20 subjects (5 nonsmokers, 6 smokers, and 9 ex-smokers) for at least 3 years. According to a balanced-block randomization method, ten patients received active treatment and ten received placebo. Either drug or placebo was to be taken as one capsule daily the first 10 days of 3 consecutive months. Each capsule of the active product contained 7 mg of a BE obtained from eight different bacterial strains. On entry (T0) and 90 days after beginning of treatment (T90), all patients underwent bronchoalveolar lavage (BAL) and peripheral blood withdrawal to assay BAL fluids and serum samples for immune parameters. The BAL recoveries, cellularity, cell differentials, and lymphocyte subsets (CD19, CD3, CD4, CD8) did not show significant differences. IgG/albumin and IgA/albumin values were not significantly different, but IgA/albumin was significantly increased in the treatment (T0 = 0.14, 0.01 to 0.27, median and range, T90 = 0.15, 0.08 to 0.45, p = 0.028) vs the placebo group when data from current smokers were excluded. Functional tests on alveolar macrophages (AM) (leading front stimulated motility and superoxide anion-O2(-)-release) showed a significant increase of random migration (T0 = 10.6, 7.0 to 23.6, T90 = 13.4, 8.1 to 28.8 microns, p = 0.02) and of stimulated motility after FMLP 10(-7) M (T0 = 13.2, 8.3 to 46.4, T90 = 18.3, 8.4 to 49.6 microns, p = 0.04), a significant increase of O2- release in basal conditions (T0 = 6.0, 1.7 to 30.5 nM/10(6) AM/10', T90 = 11.1, 5.5 to 24.5, p = 0.05) and after stimulation with opsonized zymosan (T0 = 17.7, 4.7 to 35.2, T90 = 22.1, 13.8 to 53.3, p = 0.009) in the treatment group only. Data were not significantly different in the placebo group between T0 and T90. No modifications in systemic immunity were ever observed.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404102 TI - Localized inflammatory pulmonary disease in subjects occupationally exposed to asbestos. AB - In reviewing pathology materials from patients occupationally exposed to asbestos, we identified eight patients with either localized nodules in their lung or unusual pathologic changes. The chest radiographs of six patients showed isolated parenchymal nodules thought to represent primary neoplasms. In three cases, pathologic examination of these nodules showed intraluminal fibrosis and inflammation of the distal airways, a pattern of change frequently referred to as "bronchiolitis obliterans organizing pneumonitis." In each instance, asbestos bodies were present in association with the fibroinflammatory tissue. In one case, the nodule showed a desquamative interstitial pneumonitis type pattern, and asbestos bodies were present admixed with the alveolar macrophages and occasionally within their cytoplasm. In one case, the nodule was composed of nonspecific inflammation and fibrosis with focal bronchiolitis obliterans and frequent asbestos bodies scattered throughout the area of inflammation and fibrosis, and in another case, necrotizing inflammation association with Aspergillus fungal organisms was identified. Granulomatous inflammation was the dominant pulmonary pathologic change in one patient, and the other patient's lung biopsy specimen showed a diffuse lymphocyte-plasma cell interstitial pneumonitis. The cases reported suggest that asbestos may cause localized lesions in the lung that clinically and radiographically are misinterpreted as cancer and that pathologically show inflammation and fibrosis of the distal airways. In addition, our observations suggest that asbestos may cause granulomatous inflammation, a desquamative interstitial type pneumonitis, and a lymphocytic interstitial pneumonitis type pattern. Our conclusions that asbestos may cause these pathologic changes are supported by case reports in the clinical and pathologic literature, clinicopathologic studies, and by experimental studies. PMID- 8404103 TI - Postoperative complications in patients with human immunodeficiency virus disease. Clinical data and a literature review. AB - OBJECTIVE: To compare complications after and outcome from surgical procedures between patients with human immunodeficiency virus (HIV) disease and a matched control population. DESIGN: Retrospective case review. SETTING: 476-bed university tertiary care center. PATIENTS: Of 343 patients diagnosed as having HIV disease between 1981 and September 1991, 26 (7.6 percent) were Walter Reed classifications system 3B or greater and underwent a surgical procedure with general anesthesia or, in the case of 2 patients, regional anesthesia, at the study hospital. These patients were matched to 26 control patients by severity of illness according to APACHE II severity of illness score and by age, sex, race, and anesthetic regimen. MEASUREMENTS AND RESULTS: The postoperative occurrence of dysrhythmia, hypotension, hypoxia, hemorrhage, renal insufficiency, or infection was evaluated in the study and control groups. Deaths within 3 and 30 days of the procedure and duration of survival after discharge from the hospital were recorded. Frequency of complications and 3- and 30-day mortality did not differ between the 2 groups. Duration of survival after discharge from the hospital in patients with HIV disease (7.4 +/- 9.7 months) was approximately 25 percent that in control patients (30.6 +/- 35.9 months) (p = 0.02). CONCLUSIONS: HIV disease does not increase the risk of postprocedural complications, including death, up to 30 days postprocedure. Thereafter, patients with HIV disease classified by the Walter Reed System as > or = 3B may survive for a shorter time than do equally ill patients who do not have HIV disease. Thus, needed surgical intervention should not be limited based on HIV status and concern for subsequent complications. PMID- 8404104 TI - Acute exacerbation in idiopathic pulmonary fibrosis. Analysis of clinical and pathologic findings in three cases. AB - We treated three patients with idiopathic pulmonary fibrosis who had an acute clinical exacerbation. We analyzed their clinical, radiographic, therapeutic, and pathologic findings. Their initial symptoms were influenza-like illness or cough with fever, and all had leukocytosis and elevation of C-reactive protein. Infectious events were ruled out by extensive bacteriologic and serologic examination. The patients' lung injury scores progressed rapidly to severe lung injury compatible with adult respiratory distress syndrome. Findings from bronchoalveolar lavage fluid showed marked neutrophilia and elevation of albumin concentrations. All patients showed various degrees of improvement following corticosteroid therapy. Histologic findings from open lung biopsy specimens showed both usual interstitial pneumonia (UIP) and organizing acute lung injury pattern. Whether these two forms of interstitial pneumonia (UIP and acute lung injury pattern) are variants of one disease or are unrelated and also the effectiveness of corticosteroid therapy on such conditions remain to be determined by further studies. PMID- 8404105 TI - Prospective assessment of a standardized pathologic grading system for acute rejection in lung transplantation. AB - Using the recent standardization of the pathologic definitions for acute lung rejection, we prospectively evaluated 66 consecutive bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB) specimens in 32 patients after lung transplantation. Clinical indications for bronchoscopies were surveillance (n = 44), rejection (n = 18), and infection (n = 4). Bronchoalveolar lavages were obtained from the right middle lobe or lingula in single lung transplant and from both sites in double lung transplant recipients. Cytosmears for differential cell counts were performed and 400 to 500 cells were counted. Five to eight TBB specimens were taken from two different lobes and stained with hematoxylin-eosin, elastic trichrome, and silver methenamine. Sixty-four of 66 sets of biopsy specimens were satisfactory, but 3 were eliminated because of presence of cytomegalovirus cytopathic changes. Of the remaining 61, rejection was presented in 45 (74 percent): grade 1 in 23 (38 percent), grade 2 in 19 (31 percent), and grade 3 in 3 (5 percent). In 30 of 42 (71 percent) surveillance biopsy specimens, rejection was present, grade 1 in 18 (43 percent) and grade 2 or 3 in 12 (28 percent). In TBBs performed for clinical suspicion of rejection, 15 of 18 TBB specimens (83 percent) showed rejection, grade 1 in 5 (28 percent) and grade 2 or 3 in 10 (55 percent). Of four biopsies performed for suspicion of infection, one was normal and three showed rejection in addition to infection. These three were eliminated from further analysis due to the limitation of the Lung Rejection Study Group criteria in distinguishing rejection from infection. Of the 45 episodes of rejection, 24 (53 percent) occurred during the first 3 months posttransplantation, 8 (18 percent) between 3 and 6 months and 13 (29 percent) after 6 months. Percentage of BAL lymphocytosis was significantly elevated in grade 2 or 3 rejection (28 +/- 4) when compared with grade 1 (15 +/- 3) or grade 0 (10 +/- 3) (p < 0.001). Bronchoalveolar lavage lymphocytosis also correlated with severity of rejection (r = 0.6). We conclude that according to the standardized criteria of the Lung Rejection Study Group, acute lung rejection occurs more frequently than clinically suspected early and late after transplantation and that BAL lymphocytosis correlates with the presence and severity of histologically proven rejection. PMID- 8404106 TI - Resting energy expenditure. Evolution during antibiotic treatment for pulmonary exacerbation in cystic fibrosis. AB - STUDY OBJECTIVES: To compare the changes in resting energy expenditure (REE) to concomitant changes in clinical status and pulmonary function in cystic fibrosis (CF) patients during treatment for acute pulmonary exacerbation. To determine if weight loss during exacerbation in CF is related to decreased calorie intake or increased energy needs. DESIGN: Measurements of REE, pulmonary function tests, oxygen saturation, respiratory rate, maximal inspiratory pressure (MIP), white blood cell count, chest x-ray films and attribution of clinical score (ACS) on admission, mid-hospitalization, and discharge. Anthropometric measurements on admission, assessment of dietary intake and nitrogen balance upon admission and prior to discharge. SUBJECTS: Thirteen CF patients admitted for treatment of acute pulmonary exacerbation with a mean age of 11.0 +/- 7.9 (SD) years. RESULTS: From admission to discharge, REE decreased from 44.5 +/- 9.0 to 33.8 +/- 8.5 kilocalorie (kcal)/kg/d (p < 0.003). Similarly, the ACS improved from 7.5 +/- 2.0 to 4.0 +/- 2.2 (p < 0.0001); the absolute neutrophil count decreased from 10,685 +/- 6,226/microliters to 6,363 +/- 168/microliters (p < 0.005); respiratory rate decreased from 32.6 +/- 6.2 to 25.0 +/- 3.7 breaths per minute (p < 0.01); and MIP increased from 77.5 +/- 20.0 to 90.0 +/- 20.4 cm H2O (p < 0.01). In parallel, less significant improvements occurred in pulmonary function tests, oxygen saturation and chest x-ray film scores. Calorie intake was 1,893 +/- 635 and 2,054 +/- 707 kcal/d on admission and discharge, respectively (p = NS); during hospitalization, weight increased from 23.6 +/- 10.1 to 25.7 +/- 10.1 kg (p < 0.005). While carbohydrate and fat content of the diet remained essentially unchanged, a significant increase in protein intake (3.15 +/- 0.92 to 3.5 +/- 0.81 g/kg/d [p < 0.05]) and in nitrogen balance (1.8 +/- 2.5 to 5.6 +/- 2.9 g of nitrogen per day [p < 0.05]) were observed. CONCLUSIONS: In acute CF, pulmonary exacerbation, changes in REE parallel those of clinical improvements and are more sensitive than pulmonary function tests and chest x-ray films as an objective clinical correlate. Increased metabolic requirements but not decreased dietary intake are the cause of weight loss in CF patients. PMID- 8404107 TI - Norepinephrine or dopamine for the treatment of hyperdynamic septic shock? AB - STUDY OBJECTIVE: To compare the ability of dopamine and norepinephrine to reverse hemodynamic and metabolic abnormalities of human hyperdynamic septic shock. DESIGN: Prospective, double-blind, randomized trial. SETTING: An ICU in a university hospital. PATIENTS: Adult patients with hyperdynamic septic shock after fluid resuscitation. INTERVENTIONS: Patients were assigned to receive either dopamine (2.5 to 25 micrograms/kg/min) or norepinephrine (0.5 to 5.0 micrograms/kg/min). If hemodynamic and metabolic abnormalities were not corrected with the maximum dose of one drug, the other was added. MEASUREMENTS AND RESULTS: The aim of therapy was to achieve and maintain for at least 6 h all of the following: (1) systemic vascular resistance index > 1,100 dynes.s/cm5.m2 and/or mean systemic blood pressure > or = 80 mm Hg; (2) cardiac index > or = 4.0 L/min/m2; (3) oxygen delivery > 550 ml/min/m2; and (4) oxygen uptake > 150 ml/min/m2. With the use of dopamine 10 to 25 micrograms/kg/min, 5 of 16 patients (31 percent) were successfully treated, as compared with 15 of 16 patients (93 percent) by norepinephrine at a dose of 1.5 +/- 1.2 micrograms/kg/min (p < 0.001). Ten of 11 patients who did not respond to dopamine and remained hypotensive and oliguric were successfully treated with the addition of norepinephrine. CONCLUSIONS: At the doses tested, norepinephrine was found, in the present study, to be more effective and reliable than dopamine to reverse the abnormalities of hyperdynamic septic shock. In the great majority of the study patients, norepinephrine was able to increase mean perfusing pressure without apparent adverse effect on peripheral blood flow or on renal blood flow (since urine flow was reestablished). At the same time, oxygen uptake was increased. PMID- 8404108 TI - Acute lung injury at Baragwanath ICU. An eight-month audit and call for consensus for other organ failure in the adult respiratory distress syndrome. AB - To test the expanded definition of acute lung injury (ALI), we prospectively, over a period of 8 months, studied all adult ICU admissions who fitted the definition. Our study consisted of 83 patients with the adult respiratory distress syndrome (ARDS) and 60 with mild to moderate ALI. Sepsis and trauma were the most common diagnoses on admission. The overall mortality rate was 45 percent for ARDS and 38 percent in the other group. Mortality rose significantly with associated other organ failure, the incidence of which was as follows: hepatic, 39 percent; cardiac, 38 percent; hematologic, 22 percent; renal, 21 percent; neurologic, 5 percent. Sepsis syndrome eventually occurred in 73 percent and septic shock in 38 percent of all cases of ALI. We found the expanded definition a useful grading system and consider this definition of ARDS to be currently the best. There are, however, problems with the determination of lung compliance, the effect of inverse ratio ventilation, and the lack of consensus in defining other organ failure. PMID- 8404109 TI - The course of nosocomial oropharyngeal colonization in patients recovering from acute respiratory failure. AB - We sought to determine the duration of nosocomially acquired Gram-negative bacilli (GNB) oropharyngeal colonization following hospitalization for acute respiratory failure (ARF). We selected 24 inpatients recovering from ARF who had positive oropharyngeal cultures for GNB. Follow-up cultures were obtained at the time of hospital discharge, and 2 and 4 weeks afterwards. The prevalence of GNB colonization in these patients was 14/21 (67 percent) at the time of hospital discharge and 14/23 (60 percent) 2 weeks afterwards. Both rates were greater than the control population's 7/30 (23 percent, p < 0.02 and < 0.05, respectively). Four weeks after hospital discharge, the prevalence of colonization had fallen to 7/19 (37 percent) which was not significantly different from that of controls. Five of 24 subjects were rehospitalized during the follow-up period. Pneumonia was diagnosed in only two of the five and both proved to be due to pathogens other than GNB. We conclude that the prevalence of GNB oropharyngeal colonization following ARF approaches control levels within four weeks of hospital discharge. We speculate that a post-ARF patient's risk for GNB pneumonia similarly declines. PMID- 8404110 TI - The measurement of thoraco-abdominal asynchrony in infants with severe laryngotracheobronchitis. AB - Retractions of the lower ribcage (chest wall distortion [CWD]) during inspiration are frequently observed with moderate to severe respiratory disease in the infant. Laryngotracheobronchitis (LTB) results in a reversible partial airway obstruction with severe CWD. We wished to measure the motion of the chest wall during distortion to determine the changes in minute ventilation (VE) and to evaluate this clinical sign as a means of assessing disease severity. The respiratory inductance plethysmograph was used to determine the distortion of the lower chest wall, and distortion was correlated with VE, measured at the mouth, in six infants with severe LTB and ventilatory failure. As the conditions of these infants improved, the CWD decreased with decreasing transcutaneous carbon dioxide tension (tcPCO2), VE increased from 0.27 +/- 0.12 L.min-1 x kg-1 at a tcPCO2 of 64 mm Hg to 0.64 +/- 0.06 L.min-1 x kg-1 when the tcPCO2 had fallen to 28 mm Hg. Over the same change in tcPCO2, the tidal volume (VT) increased from 4.8 +/- 0.5 ml.kg-1 to 15.7 +/- 1.4 ml.kg-1. In the most severe disease state, the excursion of the chest wall (as an inductance) was -14 +/- 3 mV in severe obstruction, but increased to 75 mV +/- 4 mV with resolution of the illness. The timing and vector of movement of the abdomen and chest wall were expressed as a Lissajous figure, which is measured as a phase angle. The severity of the disease process, as determined by tcPCO2 was directly related to the phase relationship, and thus reflected both VE and VT. The severity of the CWD may be assessed rapidly by the use of Lissajous figures. PMID- 8404111 TI - Left upper extremity edema, rash, and venous varicosities in a 52-year-old man. PMID- 8404112 TI - Back pain and fever in an elderly patient. PMID- 8404113 TI - Asbestos-related pleural plaques and lung cancer. AB - The English-language literature was reviewed to evaluate a possible relationship between asbestos-related pleural plaques and lung cancer in the absence of parenchymal asbestosis. There were six cohort studies in which the comparison group was limited to unexposed persons or the general population, four lung cancer case-control studies, and three autopsy studies. Of the 13 investigations, only 3 supported the hypothesis that lung cancer risk is elevated among persons with pleural plaques over the risk in unexposed people: 2 cohort studies from the same city in England with much the same data and 1 case-control study. These three studies had the most defects in design. The other ten studies failed to confirm the hypothesis. Thus, the weight of the evidence favors the conclusion that persons with asbestos-related pleural plaques do not have an increased risk of lung cancer in the absence of parenchymal asbestosis. PMID- 8404114 TI - Handedness and sleep apnea. AB - Left-handedness is associated with shorter life span. Many of the factors contributing to this higher mortality (eg, alcohol consumption, automobile accident, smoking) are the same as in patients with sleep apnea, who also have higher mortality than nonapneic controls. The authors hypothesized that (1) there is a higher prevalence of sinistrality in patients suspected of having sleep apnea than in the general population, and (2) left-handed persons with sleep apnea have a more severe disorder than right-handed ones. These hypotheses were tested in a prospective cohort of 970 patients referred to a sleep disorders center because of suspected sleep apnea. All patients underwent nocturnal polysomnography and measurements of blood pressure. Sixty-one patients were left handed. The distribution of handedness as a function of age in this cohort was similar to that in the general population. There were 486 patients with an apnea/hypopnea index greater than 10; of these, 34 were left-handed, and 452 were right-handed. Oxygen saturation, blood pressure, age, smoking history, and body mass index were similar in both groups. Left-handed patients with sleep apnea had a significantly higher respiratory disturbance index (RDI) than the right-handed ones (52 +/- 30 vs 38 +/- 35 [p < 0.005]). Right-handed patients were distributed equally among RDI quartiles, but 41 percent of the left-handed patients were within the highest RDI quartile, compared to 12 percent within the lowest quartile. Sinistrality appears to be associated with more severe sleep apnea, which may help to explain the higher mortality seen among the left-handed persons. PMID- 8404115 TI - Chronic Obstructive Pulmonary Disease Early Intervention Trial (Lung Health Study). Baseline characteristics of randomized participants. AB - The Chronic Obstructive Pulmonary Disease Early Intervention Trial, or Lung Health Study, is a multicenter randomized clinical trial sponsored by the Division of Lung Diseases of the National Heart, Lung, and Blood Institute. The hypothesis being tested is that over a 5-year period, a comprehensive intervention program can reduce both the rate of decline in pulmonary function and the rates of respiratory morbidity and mortality in middle-aged smokers with mild to moderate airflow obstruction. The primary outcome variable of the trial is the annual rate of decline of maximum postbronchodilator FEV1. Secondary outcomes are the development of respiratory and nonrespiratory morbidity and mortality. After screening 73,694 cigarette smokers, aged 35 to 60 years, 5,887 participants were randomized into three equal groups: usual care, smoking intervention with daily use of a metered-dose inhaler with ipratropium bromide, and smoking intervention with inhalation of placebo. Eligible participants had a ratio of FEV1 to forced vital capacity (FVC) of 70 percent or less, were free of known life-limiting conditions, expressed willingness to enter the intervention program if so randomized, and gave written informed consent prior to entry into the trial. Spirometry, methacholine challenge, and questionnaires were strictly standardized within and across centers. The purpose of this report is to describe the characteristics of randomized participants at the time of entry into the study. For both sexes, three measures of lung function--average cross-sectional FEV1/FVC ratio, FEV1, and FEV1 percentage of predicted normal--showed slight downward trends for each successively older 5-year age cohort. The increase in FEV1 after isoproterenol was 15 percent or more in only 2.4 percent of men and 2.8 percent of women. A positive response to methacholine (defined as a fall in FEV1 of > 20 percent from baseline at concentrations up to 25 mg/ml) occurred in 63 percent of men and 87 percent of women. The cross-sectional prevalences of cough, phlegm, wheeze on most days or nights, and shortness of breath were 49 percent, 43 percent, 32 percent, and 43 percent, respectively. Respiratory symptoms were reported by a higher proportion of participants in the younger age groups than in the older age groups. Participants who reported cough, phlegm, and/or wheeze averaged lower FEV1 percent predicted and higher probability of positive response to methacholine than participants who did not. Shortness of breath appeared to be significantly associated with lower lung function and higher reactivity in men but not in women.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404116 TI - Lower risk and higher yield for thoracentesis when performed by experienced operators. PMID- 8404117 TI - Severe pulmonary hypertension associated with diastolic left ventricular dysfunction. PMID- 8404118 TI - Ventricular septal rupture after myocardial infarction. Detection by transesophageal echocardiography. AB - Acute rupture of the interventricular septum is a relatively unusual complication following acute myocardial infarction. The echocardiographic features depicted by transthoracic echocardiography are well described. However, transesophageal echocardiographic description of a ruptured septum has not been previously reported. This brief report illustrates the transesophageal features of such a defect. PMID- 8404119 TI - Pulmonary actinomycosis. A cause of endobronchial disease in a patient with AIDS. AB - We report a case of a 47-year-old man with AIDS who presented with fever, cough and a lingular infiltrate. Flexible fiberoptic bronchoscopy revealed an endobronchial exophytic mass with extensive purulent exudate which on Gram stain and cytology from bronchial washings revealed Actinomyces infection. There was a clinical response to penicillin therapy, and on repeat bronchoscopic examination, there was a partial resolution of the endobronchial infection. To our knowledge, this is the first known case of endobronchial pulmonary actinomycosis in a patient with AIDS. PMID- 8404120 TI - Diagnosis of Pneumocystis carinii infection in HIV-seropositive patients by identification of P carinii in pleural fluid. AB - Pneumocystis carinii pneumonia (PCP) is the most common pulmonary complication of AIDS and is typically diagnosed by the identification of P carinii organisms in sputum, bronchoalveolar lavage fluid, or tissue obtained with transbronchial biopsy. We describe two HIV-seropositive patients with pleural effusions in whom the diagnosis of P carinii infection was made by examination of pleural fluid. Pleural effusions associated with PCP are very unusual but can provide a source of diagnostic material particularly in those HIV patients who have development of a spontaneous pneumothorax and require chest tube insertion. PMID- 8404121 TI - Repeated inhalation of nebulized albuterol did not induce arrhythmias in a patient with Wolff-Parkinson-White syndrome and asthma. AB - It has been assumed that in asthmatic patients with Wolff-Parkinson-White (WPW) syndrome, ablative therapy for the condition is necessary for the safe treatment of the asthma with beta 2-adrenergic drugs. The following case report illustrates that inhaled albuterol was safely administered to an asthmatic patient with electrocardiographic evidence of preexcitation, without the need of an ablative procedure. This case report is, to our knowledge, the first in the literature in which the cardiac rhythm of a patient with WPW syndrome was monitored during repeated inhalations of a beta 2-agonist. PMID- 8404122 TI - CPAP mask management of varicella-induced respiratory failure. AB - Pneumonia is a complication of primary varicella infection that occurs most commonly in adults and may lead to life-threatening respiratory failure. We report a case of varicella pneumonia with impending respiratory failure in which endotracheal intubation was averted by the use of CPAP mask ventilation with a favorable outcome. PMID- 8404123 TI - False pulmonary artery catheter measurements due to the scimitar (hypogenetic lung) syndrome. Potential for iatrogenic pulmonary edema. AB - In an acute trauma patient with unrecognized scimitar syndrome, physiologic measurements used in patient management decisions were misleading due to the anatomic and physiologic anomalies of the syndrome. Pulmonary artery catheter measurements believed to reflect left atrial pressures were actually measuring central venous pressures because the catheter was terminating in the scimitar vein. These erroneous measurements led to overly aggressive fluid resuscitation and iatrogenic pulmonary edema. The pathologic features of scimitar syndrome are reviewed, and the mechanism for potential mismanagement of patient volume status created by aberrant pulmonary hemodynamics is discussed. PMID- 8404124 TI - Melioidosis pneumonia and blast injury. AB - We present the case of a 24-year-old woman with acute septicemic melioidosis resulting from inhaled infective dust during a blast injury. With appropriate antibiotic treatment and supportive therapy in the ICU, the patient made an uneventful recovery. PMID- 8404125 TI - Severe ischemic injury to the proximal airway following lung transplantation. Immediate and long-term effects on bronchial cartilage. Multi-Organ Transplant Group. AB - Ischemia due to interruption of the bronchial circulation has been recognized as a cause of immediate postoperative anastomotic dehiscence in lung and heart-lung transplant recipients. Since patients do not ordinarily survive such major ischemic insults, the long-term effects of airway ischemia and the differentiation of these effects from those of transplant rejection and infection have not been clearly defined. We describe a patient who suffered extensive airway ischemia, necrosis, and subsequent diffuse airway stenosis. Loss of the bronchial circulation with variable ischemia may be a major cause of late airway abnormality responsible for significant morbidity and mortality in transplant recipients. PMID- 8404126 TI - Diagnosis of pulmonary vein varix by transesophageal echocardiography. AB - A pulmonary vein varix is a localized dilatation of a pulmonary vein that usually is asymptomatic, presenting as a mass on a chest roentgenogram. Pulmonary angiography has been the mainstay of diagnosis. This case is the first in which transesophageal echocardiography was used to diagnose pulmonary vein varix. PMID- 8404127 TI - Massive pulmonary hemorrhage complicating mitral prosthetic valve obstruction. AB - A case is reported of prosthetic mitral valve obstruction in a patient who had had three hospital admissions with massive hemoptysis. A diagnosis was finally made after two-dimensional echocardiography with Doppler and cardiac catheterization were performed. The patient was successfully treated with re replacement of the valve. Any new pulmonary or cardiac symptom in a patient with a prosthetic valve requires prompt evaluation and treatment. PMID- 8404128 TI - The use of thoracoscopy in the treatment of iatrogenic esophageal perforations. AB - Treatment options for patients with iatrogenic esophageal perforations are multiple and remain a source of controversy. Therapeutic options range from conservative, non-operative treatment to esophageal exclusion or esophagectomy. We present herein what we believe to be the first described repair of an esophageal perforation using thoracoscopic surgical techniques. We believe this approach offers several advantages over open thoracic surgery and has a role in minimizing postoperative morbidity in an already potentially life-threatening condition. PMID- 8404129 TI - Transesophageal color flow Doppler features of aorta to right ventricle fistula. AB - A patient developed aortic-to-right ventricular fistula after aortic valve replacement surgery and infective endocarditis. The fistula was not detected by transthoracic two-dimensional echocardiography and color flow Doppler. The transesophageal study, however, was instrumental in diagnosing this entity and prompting surgical resection of the fistula. PMID- 8404130 TI - Photodynamic therapy as an alternative treatment for surgery in a patient with lung cancer undergoing bone marrow transplantation. AB - We describe a patient who suffered from a bacterial pneumonia and had a left sided infiltrate on his chest radiograph. He was found to be cytopenic and acute myeloid leukemia was diagnosed. A complete remission was achieved after chemotherapy, and the patient was scheduled to have autologous bone marrow transplantation. Bronchoscopy was performed because of persistent hemoptysis and a squamous cell carcinoma in the right upper lobe bronchus was found. This small tumor was successfully treated with photodynamic therapy preventing any delay in the treatment of his leukemia, which would have occurred if surgery had been the treatment of choice. The patient is still in complete remission after a follow-up period of 12 months. PMID- 8404131 TI - An uncommon case of brainstem tumor with selective involvement of the respiratory centers. AB - We report a clinical case of a young man with a brainstem tumor with a stable alveolar hypoventilation syndrome as the only symptom of the disease. The ventilatory response to CO2 was almost absent and the ventilatory pattern during tidal breathing was very irregular. The diagnosis was made by magnetic resonance imaging of the brain and confirmed by a stereotactic brain biopsy specimen. PMID- 8404132 TI - Dextroposition of the left lower lobe after heart-lung transplantation. AB - Immediately after heart-lung transplantation for cystic fibrosis, a patient had development of a right lower lobe retrocardiac density that persisted on all postoperative chest radiographs. A computed tomographic examination of the thorax performed 3 weeks after surgery showed that there was partial collapse of the left lower lobe in the right hemithorax. The patient required a posterolateral thoracotomy for cure. PMID- 8404133 TI - Primary sternal osteomyelitis presenting as a pleural-based mass. AB - Osteomyelitis in uncommon locations can present unusual diagnostic difficulties. A patient with primary sternal osteomyelitis who presented with pain over the right supraclavicular area and a radiologic picture of a pleural-based right upper lung mass is discussed. A triple-phase bone scan was consistent with the diagnosis, and a needle aspiration of the mass revealed a staphylococcal abscess. Percutaneous drainage of the contiguous abscess and a prolonged course of antibiotic therapy cured the infection. PMID- 8404134 TI - Hydropneumomediastinum and bilateral hydropneumothorax as delayed complications of central venous catheterization. AB - A case of hydropneumomediastinum and bilateral hydropneumothorax from a central venous catheter is described. The complication is related to the position of the catheter after placement through the left internal jugular vein. Proper placement of a central venous line into the superior vena cava is essential to avoid this serious complication. PMID- 8404135 TI - Chylous pleural effusion associated with primary lymphedema and lymphangioma-like malformations. AB - We describe a patient with a chylous pleural effusion associated with primary lymphedema of his right leg and abdominal wall. On evaluation a generalized, severe hypoplasia of the lymphatic system turned out to be associated with hyperplastic, lymphangioma-like malformations. PMID- 8404136 TI - Cytomegalovirus pneumonitis. An unusual cause of pulmonary nodules in a patient with AIDS. AB - A patient with AIDS and a history of non-Hodgkin's lymphoma developed a generalized illness associated with the appearance of multiple pulmonary nodules on a chest radiograph. Cytomegalovirus infection was demonstrated by needle aspiration cytology. The patient's symptoms and radiographic abnormalities resolved completely on ganciclovir therapy. This unusual case (1) broadens the differential diagnosis of nodular pulmonary disease in patients with AIDS and (2) suggests that cytomegalovirus can cause clinically significant lung disease which may respond to standard antiviral therapy in patients with AIDS. PMID- 8404137 TI - Safety of thoracentesis in mechanically ventilated patients. AB - As practicing intensivists, we frequently diagnose pleural effusions in mechanically ventilated patients and routinely perform thoracentesis even when the patient is on positive end-expiratory pressure (PEEP). In light of a recent report, we have reviewed our experience. It is of interest that all thoracenteses were performed with patients in the lateral decubitus position. PMID- 8404138 TI - Variations in thoracic isomerism. PMID- 8404139 TI - Variability in jet nebulizer output. PMID- 8404140 TI - Role of fiberoptic bronchoscopy in diagnosis of pulmonary tuberculosis in patients at risk for AIDS. PMID- 8404141 TI - Ovarian hyperstimulation presenting as acute hydrothorax in early intrauterine pregnancy. PMID- 8404142 TI - Response of pulmonary nocardiosis to ceftriaxone in a patient with AIDS. PMID- 8404143 TI - Ventilatory criteria for systemic inflammatory response syndrome. PMID- 8404144 TI - Pneumothorax during pulmonary tuberculosis in an HIV-infected patient. PMID- 8404145 TI - Recurrent interstitial pneumonitis and dexfenfluramine. PMID- 8404146 TI - Mycoplasma infection mimicking hypersensitivity pneumonitis. PMID- 8404147 TI - Pulmonary artery perforation caused by a flow-directed balloon-tipped catheter. PMID- 8404148 TI - Chylothorax. A new surgical strategy. PMID- 8404149 TI - Atrial fibrillation complicating transesophageal echocardiography. PMID- 8404150 TI - Ice fishing as a risk factor for pulmonary emboli. PMID- 8404151 TI - Expectoration of an occult foreign body six asymptomatic years after aspiration. PMID- 8404152 TI - A historical footnote to respiratory therapy. PMID- 8404153 TI - Activation of bone marrow eosinophils in asthma. PMID- 8404154 TI - Elevated levels of carcinoembryonic antigen and cancer antigen 125 in a thymic cyst. PMID- 8404155 TI - Ultrasonographic detection and guided biopsy of thoracic osteolysis. AB - Eighty-six osteolytic lesions of the rib cage were examined by means of conventional radiography and ultrasonography. The ultrasonographic criteria of osteolysis are described. Only 1 rib metastasis was missed by ultrasonography while 13 lesions were not detected by x-ray film. Of 63 ultrasonographically guided biopsies, 62 resulted in the final diagnosis; complications were not observed. The results recommend ultrasonographic examination combined with guided biopsy as the first diagnostic step, if a metastasis of the rib cage is suspected after physical examination or after a pathologic bone scan. PMID- 8404156 TI - The role of transcarinal needle aspiration in the staging of bronchogenic carcinoma. AB - Metastatic spread to subcarinal lymph nodes in patients with bronchogenic carcinoma generally indicates unresectability. Transcarinal needle aspiration of the main carina (TCNA) has been used to obviate the need for more invasive procedures, particularly thoracic surgery. Of 510 transbronchial needle aspirations performed at our institution from 1983 to 1991, 88 (17 percent) were from the main carina in patients with bronchogenic carcinoma. We reviewed these 88 TCNA procedures to assess our experience with TCNA in the staging of lung cancer. The TCNA results were positive in 32 of 88 (36 percent) patients (20 non small-cell cancers, 12 small-cell lung cancers). Following bronchoscopy, TCNA was the only evidence of unresectability in all 20 patients with non-small-cell lung cancer and was the only mode of diagnosis in 5 of 12 (42 percent) patients with small-cell lung cancer. Thirteen patients with non-small-cell lung cancer and positive TCNA also had positive bronchial secretion cytologic studies. Five of these patients had further subcarinal sampling and in all cases metastatic involvement was confirmed. TCNA was positive in 29 of 67 (43 percent) patients with radiographic evidence of mediastinal adenopathy. Of the remaining 38 patients with radiographic evidence of mediastinal adenopathy and negative TCNA results, 23 patients had further mediastinal sampling with mediastinoscopy or thoracotomy and in all cases mediastinal spread of cancer was established. Nineteen of 58 (33 percent) patients with an endoscopically normal-appearing main carina had a positive TCNA, while 13 of 30 (43 percent) patients with broadening or widening of the main carina had positive TCNA results. There were no complications. We conclude that TCNA is often a safe and useful staging modality in patients with bronchogenic carcinoma. PMID- 8404157 TI - Pneumothorax with fine-needle aspiration of thoracic lesions. Is spirometry a predictor? AB - To assess the value of spirometry for predicting the risk of pneumothorax (PTX) following percutaneous fine needle aspiration (FNA) of thoracic lesions, we examined retrospectively the incidence of PTX in 89 FNA and associated spirometry. Spirometry results were classified as normal, obstructed, or restrictive. Overall, the PTX rate was 20 percent. When the PTX occurrence was analyzed based on our spirometry classification, no significant difference was found between the groups. A PTX occurred in 27.8 percent of the FNA performed in patients with normal spirometry. On further analysis of specific spirometry measurements (FEV1, FVC, FEV1 percent predicted, and FEV1/FVC) and incidence of PTX, no significant correlation in PTX rates was found. These data suggest that the FNA pneumothorax is not correlated with lung function as measured by routine spirometry. PMID- 8404158 TI - The utility of fiberoptic bronchoscopy in the evaluation of the solitary pulmonary nodule. AB - To determine the value of routine, preoperative, fiberoptic bronchoscopy (FB) for diagnosing and treating patients (pts) with solitary pulmonary nodules (SPNs), we retrospectively reviewed the records of all pts with SPNs undergoing FB at Walter Reed Army Medical Center between January 1986 and December 1989. We defined SPNs radiographically as < or = 6 cm peripheral pulmonary lesions completely surrounded by pulmonary parenchyma. Of 191 charts reviewed, 91 (72 bronchogenic carcinomas [BC], 7 carcinoid tumors, 12 benign) constitute the study population. Fifty-four charts were eliminated because preoperative, clinical-radiologic staging revealed advanced (greater than stage I) BC or extrathoracic malignancy metastatic to the lung (44), the clinicians suspected benign disease and elected medical followup (3), the pt had medically inoperable disease (3), or the pt refused surgery (4). Forty-six charts were incomplete or unavailable. Fiberoptic bronchoscopy revealed one unsuspected vocal cord carcinoma and no occult synchronous BCs. Five pts had submucosal or endobronchial tumors and biopsy specimens showed BC in four of five tumors from which specimens were taken. Four of 66 (6 percent) cytologic evaluations of bronchial brushings or washings diagnosed BC. In pts shown at surgery to have BC, 9 of 30 transbronchial lung biopsy (TBBx) specimens showed BC. Diagnostic yield of TBBx specimens was not improved in the pts who underwent biopsies under fluoroscopic guidance. The 16 FB specimens positive for BC concurred 100 percent with the surgical specimens. The FB findings did not obviate the need for surgery nor alter the surgical stage of BC. A preoperative diagnosis of malignancy did not affect operative time or operative procedure, because many pts required frozen-section biopsy of mediastinal lymph nodes prior to lung resection. At our institution, routine, preoperative FB did not measurably benefit pts with SPNs. PMID- 8404159 TI - Complications of fiberoptic bronchoscopy in thrombocytopenic patients. AB - STUDY OBJECTIVE: To determine the risk of epistaxis and pulmonary hemorrhage due to fiberoptic bronchoscopy (FOB) and bronchoalveolar lavage (BAL) in the presence of thrombocytopenia. DESIGN: Prospective study of all patients undergoing FOB with BAL with a 4.9-mm-diameter bronchoscope after bone marrow transplantation (BMT) during a 6-month period. SETTING: A single BMT center. PATIENTS: Forty seven BMT recipients undergoing 66 FOB with BAL. Thrombocytopenia (platelets < 100,000/ml) was present in 58 (88 percent). Platelets were < 50,000/ml in 44 (67 percent) and < 20,000/ml in 13 (20 percent). In the thrombocytopenic patients, FOB with BAL was transnasal in 37 (64 percent), transoral in 5 (9 percent), and via endotracheal tube in 16 (28 percent). INTERVENTIONS: Fiberoptic bronchoscopy with BAL using a bronchoscope (Pentax FB-15H) (4.9-mm diameter). In one case, a pediatric bronchoscope (Pentax FB-10H; 3.5-mm diameter) was used in a 7-year-old patient. MEASUREMENTS AND RESULTS: The BAL was diagnostic in 22 of 47 patients studied (47 percent). Complications occurred in 7 of 58 (12 percent) thrombocytopenic patients (epistaxis and/or hemoptysis, 4; bradycardia, 2; bronchospasm, 1) of which all but 1 were minor and self-limiting. One life threatening complication of severe epistaxis occurred during a transoral FOB in a patient with prior epistaxis (platelet count, 18,000/ml). One of 8 (13 percent) nonthrombocytopenic patients had hemoptysis. No patient had worsening fever or oxygenation at 4 h and no patient displayed worsening radiographic infiltrates suggestive of pulmonary hemorrhage attributable to the BAL at 24 h. CONCLUSIONS: We conclude that transnasal FOB in thrombocytopenic patients was safe, being associated with minor airway bleeding in 3 of 37 patients (8 percent). In conclusion, FOB with BAL, even via the transnasal route, may be performed with relative safety despite the presence of significant thrombocytopenia. PMID- 8404160 TI - A comparative evaluation of propofol and midazolam as sedative agents in fiberoptic bronchoscopy. AB - Propofol, a new intravenous sedative agent, was investigated in 41 asthmatic patients undergoing day-case (outpatient) fiberoptic bronchoscopy. The study design was a randomized comparison between propofol and midazolam, which is a well-established intravenous sedative agent. The age, weight, and American Society of Anesthesiologists physical status and lung function of the two groups were not significantly different. Mean (SD) induction dose of propofol was 104.7 (30.1) mg with a maintenance dose of 121.9 (38.5) mg. Corresponding values of midazolam were 9.3 (3.1) mg and 3.7 (2.3) mg. The required level of sedation was achieved significantly faster with propofol, mean (SD) 125.4 (39.8) s, compared with midazolam, 179.4 (55.2) s (p < 0.001). Significantly faster recovery was noted with propofol compared with midazolam in terms of time to recall name and date of birth 2.3 (1.7) min vs 6.3 (8.6) min, (p < 0.045). Alertness scored with the digital symbol substitution test (DSST) returned to prebronchoscopy values in the propofol group at 30 min, DSST score = 35.9 (18.2) vs 13.4 (9.1), in the midazolam group (p < .0001) and was still significantly higher at 90 min-39.4 (17.9) and 23.1 (13.8) (p < 0.01). We conclude that propofol is a useful sedating agent in fiberoptic bronchoscopy with similar efficacy to midazolam but with a faster onset of action and a more rapid recovery. These represent significant advantages for day-case procedures. PMID- 8404161 TI - General vs local anesthesia. Effect on bronchoalveolar lavage findings. AB - Bronchoalveolar lavage (BAL) can be performed with the patient undergoing either local or general anesthesia (GA). This study investigates whether the type of anesthesia affects BAL fluid and cell recovery. Eighty patients, were selected for study. Fluid recoveries were significantly less in the GA group for both the bronchial and alveolar lavages. The differences were confirmed for BAL fluid recovery in a subsequent group of 120 unselected patients. Bronchoscope size did not appear to affect recovery, nor did anesthesia time; BAL fluid recovery from patients with respiratory failure who were intubated and mechanically ventilated was similar to that in the GA group, suggesting that lower recovery rates may be due to mechanical ventilation. The BAL fluid cell counts were related to fluid recovery, but airway neutrophils represented a higher percentage of BAL lavage fluid cells in the GA lavages, independent of differences in the volume of lavage fluid recovered. PMID- 8404162 TI - Fibrogenic activities in bronchoalveolar lavage fluid of farmer's lung. AB - Hyaluronic acid (HA), type III procollagen, fibronectin, and fibroblast growth factors (FGF) were measured in 43 bronchoalveolar lavage fluid (BALF) specimens obtained from 38 patients with farmer's lung (FL) and in BALF of 9 nonexposed normal control subjects. Bronchoalveolar lavage was done in 21 farmers with acute FL (acute) and in 22 with a history of previous FL (Ex) who were still in daily contact with dairy barns. All farmers from the acute and Ex groups had a lymphocytic alveolitis, respectively, 62.7 (3.5) percent (mean [SEM]) and 48.1 (4.3) percent. Hyaluronic acid, type III procollagen, fibronectin, and FGF were all highly increased in acute disease. These substances were also increased in the BALF of subjects of the Ex group who had no clinical symptoms or signs of acute disease at the time of lavage, but were actively farming. The increase in type III procollagen, however, was less in this group than in the subjects with acute disease. These observations suggest that the fibrosing activities and potentialities of the allergic alveolitis of FL are fully expressed at the time of clinical presentation and also in the subclinical phase of the disease in susceptible farmers who remain exposed after an initial acute phase of the disease. PMID- 8404163 TI - Bronchoalveolar lavage and transbronchial biopsy in children following heart-lung and lung transplantation. AB - Between July 1985 and March 1992, 20 children received either heart-lung (11), double lung (8), or single lung (1) transplants at our center. Since 1988, flexible fiberoptic bronchoscopy with bronchoalveolar lavage and transbronchial biopsy have been carried out to monitor for rejection or infection in these patients. As of March 31, 1992, we have performed a total of 112 transbronchial biopsies in our patients, who ranged from 6.8 to 18 years of age and 19.3 to 67.3 kg in weight. All but two of these procedures were carried out using conscious sedation and a transnasal approach. Four to seven biopsy samples were obtained at each procedure. One patient had hemorrhage (< 100 ml) and no patient had pneumothorax as a complication. Of the biopsy samples, 72.4 percent had a surface area of greater than 2 mm2, and 89.5 percent of the biopsy samples were deemed adequate for pathologic interpretation. We believe that for the majority of pediatric lung or heart-lung recipients, flexible bronchoscopy and transbronchial biopsy using conscious sedation and a transnasal approach is safe and permits the recovery of adequate tissue for pathologic evaluation. The avoidance of general anesthesia, endotracheal intubation, and mechanical ventilation at the time of bronchoscopy and transbronchial biopsy probably decreases the likelihood of pneumothorax as a complication of the procedure. PMID- 8404164 TI - Bronchogenic carcinoma in patients seropositive for human immunodeficiency virus. AB - As the HIV epidemic continues and the patients are closely followed throughout the course of the illness from HIV seropositivity to depressed total CD4 counts, the natural history of lung cancer in this population is evolving. HIV-infected patients with lung cancer are in general younger men with significant smoking histories. Adenocarcinoma is the predominant cell type. There has been no correlation between stage of lung cancer and CD4 counts. The lung cancer stage at presentation has also not affected prognosis (no survivors beyond 1 year from diagnosis). While HIV seropositivity has not yet been identified as a risk factor for bronchogenic carcinoma, the current literature suggests that lung neoplasms behave in an aggressive manner in HIV-positive patients. We present two cases to illustrate the value of transbronchial biopsy which should be performed in all patients with masses, nodules, or focal lesions that persist despite appropriate therapy for opportunistic organisms in HIV-positive patients. PMID- 8404165 TI - Septic shock due to toxoplasmosis in patients infected with the human immunodeficiency virus. AB - PURPOSE: To describe the presentation and clinical course of septic shock due to Toxoplasma gondii in patients infected with the human immunodeficiency virus (HIV). PATIENTS AND METHODS: From April 1988 to February 1992, nine HIV-infected patients were admitted because of predominant septic shock (7 patients) or developed septic shock in the ICU (2 patients). The recent CD4+ cell count ranged from 2 to 84 x 10(6)/L. RESULTS: The main clinical features were (1) a history of fever for longer than 15 days, with a recent increase to more than 39.5 degrees C; (2) a recent history of dyspnea (< 15 days, 8 cases; < 7 days, 3 cases); and (3) recent onset of thrombocytopenia (6 of 9 cases). All patients were in shock (hyperkinetic profile in 6 of 7; hypokinetic in 1 of 7), and 8 of 9 were in respiratory distress (ratio of PaO2 over fractional concentration of oxygen in the inspired gas of 117 +/- 23; range, 88 to 155). Chest roentgenograms revealed diffuse alveolar infiltrates in six of nine cases. The serum lactate dehydrogenase (LDH) activity was 6,510 +/- 5,080 IU/L (range, 1,010 to 15,450 IU/L). Serologic tests for T gondii were negative in two cases. Toxoplasma gondii was isolated from lung (9/9), bone marrow (5/7), or blood (2/2). One, 3, and 2 patients had brain, ocular, and myocardial involvement, respectively. No other microbial pathogens were isolated. Seven patients died, 5 less than 3 days after admission. CONCLUSION: Disseminated toxoplasmosis can cause septic shock in HIV infected patients. In two cases, the disease was probably a primary infection. The association of high fever, acute dyspnea, recent onset of thrombocytopenia, and a very high level of LDH activity is suggestive of disseminated toxoplasmosis. PMID- 8404166 TI - Cardiac tamponade in patients infected with HIV. A report from an inner-city hospital. AB - Thirty-seven consecutive cases of cardiac tamponade occurring over a 6-year period were retrospectively studied from January 1986 to December 1991 in an inner-city public teaching hospital. All episodes were secondary to medical illnesses. Thirteen (35 percent) of 37 patients had HIV infection. Significant differences (HIV vs non-HIV) in clinical presentation were noted in the following parameters: (1) age (34 +/- 7 years vs 56 +/- 14 years, p < 0.001); (2) febrile presentation (62 percent vs 17 percent, p < 0.02); and (3) presence of pulmonary infiltrates (54 percent vs 17 percent, p < 0.03). All but two patients underwent pericardiocentesis or had operative creation of a pericardial window. Two patients had purulent pericarditis; two patients had tuberculous pericarditis. In the remaining patients, there was no evidence of opportunistic infection or malignancy based on cultures of pericardial fluid and histopathologic analysis of tissue. Six of 13 patients with HIV infection survived to be discharged from the hospital following hospitalization for the illness. We conclude the following: (1) HIV infection is frequently found in patients with cardiac tamponade at inner city hospitals; (2) when young patients present with cardiac tamponade, the coexistence of fever and pulmonary infiltrates is suggestive of underlying HIV infection; and (3) the etiology of the pericardial effusion is not confirmed in the majority of patients with HIV infection. PMID- 8404167 TI - Surgical myocardial revascularization after acute infarction. AB - The records of 288 patients undergoing isolated surgical myocardial revascularization between June 1989 and September 1992 were reviewed to determine the relative risk associated with surgery after an acute myocardial infarction (MI). A total of 73 patients (25 percent) were operated on within 30 days of an acute infarction while 215 patients (75 percent) had no history of recent infarction. Patients with an acute infarction were more likely to have regional wall motion abnormalities on ventriculography (mean wall score 6.7 vs 4.9, p = 0.001), require preoperative balloon pumping (15.1 percent vs 5.6 percent, p = 0.01), and have recent symptoms of congestive heart failure (23 percent vs 12 percent, p = 0.02). Patients with an acute MI also had higher NYHA functional classification and greater urgency of surgery. Despite these differences, overall mortality was lower in the acute MI group than in the control population (1.4 percent vs 2.3 percent, p = 0.623). Weaning from bypass was not appreciably more difficult in patients with an acute MI, nor were there differences in the mean number of hours of balloon pump or inotrope support. PMID- 8404168 TI - Cardiac arrhythmias after inhaled bronchodilators in patients with COPD and ischemic heart disease. AB - Selective beta 2-agonist aerosols may produce significant cardiovascular effects. In the present study we used Holter monitoring to compare the arrhythmogenic effects of inhaled terbutaline (TE), a beta 2-agonist, with that of ipratropium bromide (IB), a nonabsorbable cholinergic drug. Fourteen patients with concomitant obstructive lung disease, ischemic heart disease, and complaints of postinhalation palpitations were studied in a random, double-blind, cross-over fashion. Both drugs significantly improved vital capacity and FEV1. Heart rate and the frequency of premature beats were not significantly affected by the bronchodilators. We conclude that no clear connection between inhaled bronchodilators and arrhythmias could be demonstrated. PMID- 8404169 TI - Unusual clinical presentations of secundum atrial septal defect. AB - Atrial septal defect (ASD) is one of the most common congenital cardiac anomalies found in the adult population. Although usually asymptomatic in childhood, ASD will be symptomatic in approximately 75 percent of adults. The most common symptoms include fatigue, dyspnea on exertion, and palpitations. However, the presentation of ASD can be protean. We present four patients with secundum ASD with unusual clinical manifestations. Patient 1 had moderately severe mitral regurgitation. Patient 2 had pulmonary edema with generalized left ventricular impairment. Patient 3 had chest pain typical of angina pectoris. Patient 4 had right-to-left shunt following an orthopedic surgical procedure. These patients had chest radiographs and electrocardiograms typical of secundum ASD, but their presentations were uncommon. In three of four of these patients, dramatic resolution of symptoms followed surgical repair of their ASD. PMID- 8404170 TI - Pathogenesis of Cheyne-Stokes respiration in patients with congestive heart failure. Relationship to arterial PCO2. AB - In order to determine which patients with congestive heart failure (CHF) develop Cheyne-Stokes respiration (CSR) during sleep, we compared the cardiorespiratory profiles of CHF patients with CSR to those of CHF patients without CSR. Overnight polysomnography and continuous transcutaneous PCO2 (tc PCO2) monitoring, estimation of left ventricular ejection fraction (LVEF), pulmonary function tests, and chest radiograph were performed on 16 consecutive patients with chronic, stable CHF. The tc PCO2 monitor (Kontron 7640) was calibrated so that measurements reflected arterial PCO2 values. A mean value was calculated for wakefulness (W) and total sleep time (TST). Circulation time (CT) from the lung to the carotid body was estimated from the end of an apnea or voluntary breath hold to the nadir of oxygen desaturation recorded on an ear oximeter. The duration of CSR was expressed as a percent of TST. Nine patients developed CSR during sleep (52.5 +/- 31.6 percent TST) (group 1) and 7 did not (group 2). All patients were male and both groups were a similar age (64 +/- 8 vs 63 +/- 4 years) and weight (body mass index, 28.1 +/- 3.5 vs 25.4 +/- 3.4 kg/m2). There were no significant intergroup differences between LVEF (22 +/- 5.2 vs 24.1 +/- 5.2 percent), CT (19.1 +/- 3.6 vs 15.9 +/- 6.7 s), SaO2 (W) (94 +/- 1.2 vs 92.4 +/- 2.1 percent), and SaO2 (TST) (90.8 +/- 2.7 vs 92.4 +/- 2.1 percent). The tc PCO2 (W) was lower in group 1 (34.4 +/- 3.5 vs 38.1 +/- 1.9 mm Hg), increased during sleep by a similar amount in both groups (1.6 +/- 1.5 vs 2.1 +/- 2.2 mm Hg), and was significantly lower during sleep in group 1 (36.1 +/- 3.4 vs 40.2 +/ 2.2 mm Hg). We conclude that CHF patients with CSR hyperventilate during sleep and wakefulness and that CHF patients with awake hypocapnia are more likely to develop CSR during sleep. These findings indicate that arterial PCO2 is important in determining which CHF patients develop CSR. PMID- 8404171 TI - Pulmonary diffusion abnormalities in heart transplant recipients. Relationship to cytomegalovirus infection. AB - Lung function of patients with heart failure is characterized by a variety of changes proposed as being due to passive congestion, secondary pulmonary fibrosis, and/or recurrent pulmonary emboli. A diffusion impairment thought to be due to cyclosporine has also been noted in patients following heart transplantation. Similar changes of unclear origin have been observed in renal transplant recipients. The objective of this study was to determine the extent to which lung function changes are reversible by cardiac transplantation and relate changes to the status of the recipients lung in the presence of possible vascular, iatrogenic, immune, or infectious injury. We analyzed the data of 22 patients who underwent lung function testing before and after heart transplantation and correlated changes to hemodynamic change, episodes of rejection, concentration of cyclosporine, and cytomegalovirus infection. Despite excellent graft function, the carbon monoxide transfer factor deteriorated to a mean of 57 percent of predicted postoperatively. The fall in diffusion factor did not correlate with episodes of cardiac rejection, cyclosporine levels, or hemodynamic status. In those patients who had serologic evidence of cytomegalovirus infection, the reduction in transfer factor was greater compared to those without infection despite a normal chest radiograph. The effects of cardiopulmonary bypass were unlikely to have been responsible for the abnormalities as lung function was assessed at a mean of 14 months after surgery. In heart transplant recipients, a change in diffusion capacity may represent an additional marker for cytomegalovirus infection and reflect infectious/immune injury late following surgery. PMID- 8404172 TI - Obstructive sleep apnea in heart transplant patients. A report of five cases. AB - Clinically significant obstructive sleep apnea syndrome (OSAS) was diagnosed in five patients who had undergone heart transplantation and they represent 2.5 percent of our transplantation series. To determine if these individuals had unique clinical features that would have suggested the presence of OSAS, we reviewed their case histories. Although four of five patients had symptoms of OSAS prior to transplant, none was suspected of having the diagnosis before their surgery. Excessive daytime sleepiness and loud snoring were noted in all patients, and there were no unusual clinical features that characterized these individuals. Our data indicate that the occurrence of OSAS in heart transplant recipients approximates the prevalence in the general population. Because OSAS may adversely affect cardiac function, we recommend that heart transplantation candidates be screened for a history suggestive of OSAS, and that polysomnography be performed if it is present. PMID- 8404173 TI - Acoustic analysis of vowel emission in obstructive sleep apnea. AB - We studied vocalization in 18 men with obstructive sleep apnea syndrome (OSAS) (age, 49 [7.5] years; body mass index [BMI] 33.6 [7.6]) and 10 normal men as a control group (age, 46.7 [6.2] years; BMI 24.6 [2.2]). Polysomnographic data for patients with OSAS were as follows: total sleep time (TST), 387.5 [27.9] min; awake, 17.6 (12.6% TST); stage 1, 19.8 (18.7 percent TST); stage 2, 54.8 (23.2 percent TST); stage 3 and 4, 1.5 (0.3 percent TST); and stage REM, 4.2 (1.7 percent TST). Apnea hypopnea index (AHI) was 43.0 (18.2) and lowest O2 saturation was 73.6 (11.4). We recorded the following sounds in all subjects: /a/ as in "father"; /e/ as in "get"; /i/ as in "see"; /o/ as in "go"; /u/ as in "too." Three maneuvers for each vowel sound were taken for analysis. Signals were digitized at 10,000 Hz. Fast Fourier transformation was applied to segments of 512 points of each utterance corresponding to the vowel sound. The following parameters were obtained: maximum frequency of harmonics, mean frequency of harmonics, and the number of harmonics. RESULTS: There were significant differences between both groups in the maximum frequency of harmonics of /i/ and /e/ vowels. (For /i/: 2,650 [672] Hz controls; 425 [71.2] Hz OSAS. For /e/: 2,605 [772.3] Hz controls; 1,250.0 [828.4] OSAS). The number of harmonics for /i/ vowel was 4.5 (1.2) for controls as compared with 2.7 (1) Hz for OSAS. CONCLUSIONS: Vocalization in patients with OSAS is different from normal subjects. Vowel /i/ can distinguish these patients from normal subjects. PMID- 8404174 TI - The ventilatory response to arm elevation of patients with chronic obstructive pulmonary disease. AB - Although arm activity is poorly tolerated by patients with COPD, the ventilatory response to arm elevation alone is not well understood. We therefore studied the ventilatory response to arm elevation using a customized arm support sling to eliminate the effect of an increase in metabolic activity that might be attributable to independent arm elevation and used leg exercise to increase metabolic activity. During arm elevation at rest, there was a significant decrease in vital capacity (180 ml) and a small decrease in functional residual capacity (120 ml) as measured by body plethysmography. Minute ventilation was unchanged. When supported arm elevation (SAE) was compared with the control arm position (CAP), minute ventilation was unchanged although the pattern of breathing became more rapid and shallow (mean +/- SD, SAE vs CAP: fb = 17.9 +/- 5.3 vs 16.2 +/- 4.8 breaths.min-1; VT = 533 +/- 126 vs 579 +/- 142 ml; p < 0.05). During steady-state leg exercise, the increase in VO2, VCO2 and VE did not differ between SAE and CAP; however, both fb and VT changed toward a more rapid, shallow pattern of breathing (SAE vs CAP: fb = 24.3 +/- 3.0 vs 22.8 +/- 3.5 breaths.min 1; VT = 990 +/- 293 vs 1,081 +/- 309 ml; p < 0.05). During unsupported arm elevation VO2, VCO2, and VE, and fb were significantly greater than during the CAP. Approaches that train arm muscles and strategies that either support arm muscles or allow for frequent rests during upper arm activity may improve the endurance and the quality of life for COPD patients. PMID- 8404175 TI - Effects of theophylline withdrawal in severe chronic obstructive pulmonary disease. AB - To determine the value of theophylline in the maintenance therapy of patients with severe chronic obstructive pulmonary disease (COPD), we conducted a trial of theophylline therapy withdrawal in 38 clinically stable patients with severe COPD (FEV1 < 60 percent) predicted. Symptoms, lung function, blood gases, and 6-min walking distance were assessed on days 1 and 2. Quality of life and overall dyspnea were scored using four different clinical rating scales. Theophylline therapy was continued in 20 patients and replaced by placebo from day 3 on in the remainder; measurements were repeated on days 5 and 6. Withdrawal of theophylline therapy resulted in significant (p < 0.05) deterioration in lung function, exercise performance, and two indices of overall dyspnea, and a significant increase in scoring of symptoms and auscultation findings. Individual analysis revealed a clinically relevant deterioration in 72 percent of patients from whom theophylline therapy was withdrawn, while only 15 percent of patients receiving theophylline exhibited deterioration. No major side effects were observed. Our data show that about half of the patients with severe COPD can be considered as theophylline responders. The response of these patients to withdrawal of theophylline therapy suggests that the clinical effectiveness of this drug cannot be explained exclusively by bronchodilation. Due to the inherent difficulties in predicting response to theophylline, its effectiveness in patients with severe COPD should be determined individually, including assessment of exercise performance and ratings of dyspnea. PMID- 8404176 TI - Inhaled beta 2-agonist and positive expiratory pressure in bronchial asthma. Influence on airway resistance and functional residual capacity. AB - INTRODUCTION: Positive expiratory airway pressure seems to dilate narrowed or collapsed airways, but this may be accompanied by a maintained and harmful increase in resting lung volume in obstructive pulmonary disease. PURPOSE: To evaluate the influence of inhaled terbutaline and positive expiratory pressure (PEP) on airway resistance (Raw) and functional residual capacity (FRC) in bronchial asthma. DESIGN: Randomized crossover design, single blind with regard to inhaled medication, open with regard to PEP (PEP can be felt). MATERIAL AND METHODS: Ten patients with bronchial asthma inhaled placebo and terbutaline in doses of 0.125 mg, 0.5 mg, and 1.5 mg by cone spacer combined with a facemask giving 0, 10, or 15 cm H2O PEP on separate days. FRC and Raw were measured by body plethysmography before and after inhalations. Data were analyzed by analysis of variance with terbutaline dose and PEP as factor levels. RESULTS: The effect of terbutaline: Raw decreased significantly (p < 0.0001) after 0.125 mg and 1.5 mg. The FRC did not change significantly. The effect of PEP: Raw decreased, but significantly only when the dose of 1.5 mg terbutaline was excluded from the analysis. Raw decreased with PEP 10 and 15 cm H2O, mean 0.6 (95 percent CI: -1.1, -0.2) and 0.9 (95 percent CI: -1.3, -0.4) cm H2O/L/s. The FRC did not change significantly with the PEP level. CONCLUSION: PEP only had influence on Raw when insufficient doses of terbutaline were inhaled, whereas once an efficient dose of terbutaline was administered, significant bronchodilation was achieved with or without PEP. Positive expiratory pressure did not increase FRC. PMID- 8404177 TI - Optimal duration of nebulized albuterol therapy. AB - The output from a jet nebulizer was analyzed for aerosol profile, solution output, and delivery of albuterol at three different initial volume fills to determine the changes that occur during the course of nebulization. Increasing diluent volume led to significantly greater delivery of the albuterol initially placed in the nebulizer. Albuterol delivery from the nebulizer ceased completely following the onset of inconsistent nebulization (sputtering) as determined audibly and by laser particle analysis. Aerosol output rate declined by one-half within 20 s of the onset of sputtering. The albuterol concentration in the nebulizer solution increased significantly once the aerosol output declined. The weight of solution delivered as determined by change in weight of the nebulizer could not be fully accounted for as aerosol volume. It appeared that this discrepancy represented loss of water by evaporation. Aerosolization past the point of initial jet nebulizer sputtering is unproductive. PMID- 8404178 TI - A statistical approach for assessment of bronchodilator responsiveness in pulmonary function testing. AB - OBJECTIVE: Because published criteria for bronchodilator responsiveness are based on population variability and compare only the single best measures before and after intervention, we hypothesized that the variability of the FEV1 and FEV3 of each patient would better determine that patient's responsiveness. DESIGN AND INTERVENTIONS: Five 3-s forced expiratory maneuvers were used for each of 3 sequential portions of the study: baseline, after nebulized saline solution, and after nebulized albuterol. SETTING: Clinical pulmonary function laboratory in a county/university teaching hospital. PATIENTS: Fifty consecutive adult patients with obstruction referred for routine testing and thought to be able to complete the study. (The primary diagnoses were found to be equally divided between asthma, bronchitis, and emphysema; equal numbers had very severe, severe, moderate, mild, or minimal obstruction.) MEASUREMENTS AND RESULTS: The primary finding was that 36 patients were found to be responders (consistent and statistically significant FEV1 and FEV3 improvement after albuterol). The 14 identified as nonresponders would have been so categorized by all other published criteria. For assessing responsiveness, we found FEV3 measures preferable to vital capacity measures, because the latter depends on the duration of the maneuver. CONCLUSIONS: The percentage of responders identified is higher than recognized by any other published criteria. Most criteria would not have identified the responders with very severe or minimal airways obstruction. PMID- 8404179 TI - Bronchial responsiveness to ultrasonic fog in occupational asthma due to toluene diisocyanate. AB - To determine the validity of ultrasonic nebulization of distilled water (UNDW, "fog") in comparison with methacholine challenge, in the assessment of toluene diisocyanate (TDI) asthma, we evaluated 75 subjects exposed to TDI with work related respiratory symptoms. Subjects were submitted to bronchial challenge with methacholine at first, thereafter to UNDW inhalation and to specific challenge with TDI. The diagnosis of TDI-asthma was made in 30 of 75 patients (40 percent) who developed a bronchoconstrictive response to the specific challenge (reactors). Sensitivity and specificity of UNDW alone, methacholine alone, and of the combination of the two tests were determined with the results of the specific challenge with TDI as the "gold standard." Both frequency and severity of bronchoconstrictive response to UNDW (FEV1 decrease > or = 15 percent) and the degree (PD15 FEV1) and frequency of bronchial hyperresponsiveness to methacholine were significantly higher in TDI reactors than in nonreactors. The UNDW had higher specificity (82.2 percent vs 51.1 percent) but lower sensitivity (40 percent vs 76.7 percent) than methacholine. The combination in parallel (positivity of any of the two challenges) of methacholine and UNDW challenge did not change sensitivity to a great extent (80 percent vs 76.7 percent), whereas combination in series (positivity of both challenges) had considerably greater specificity (86.7 percent vs 51.1 percent) than methacholine alone. We conclude that in the assessment of TDI-asthma, the validity of UNDW challenge alone is limited since it is insufficiently sensitive. Instead, combining UNDW and methacholine challenge when methacholine is positive improves our ability in identifying subjects with TDI-asthma diagnosed with the specific challenge. This procedure constitutes a first objective confirmation of a suggestive history of TDI-asthma that is useful for clinical purposes. However, especially for medicolegal purposes, the definitive diagnosis requires the specific challenge. PMID- 8404180 TI - Influence of maternal smoking on variability of peak expiratory flow rate in school children. AB - Diurnal variability of peak expiratory flow rates (PEFRs) was assessed in 1,237 children. The PEFR was measured twice daily over a 1-week period. As an index of variability, the log of a week's mean of daily amplitude was calculated. Linear regression analyses revealed a significant positive association between maternal smoking and the variability of PEFR for nonasthmatic children. For these children, exposure to maternal smoking was associated with a 13.7 percent increase (confidence interval [CI], 3.8 to 24.7 percent) in PEFR variability. For asthmatic children an effect was found for nonatopic (54.7 percent increase; CI, 5.5 to 226.8 percent) but not for atopic children (-8.5 percent change; CI, -41.2 to 42.3 percent). In the latter group, there was evidence that mothers changed their smoking habits subsequent to the development of disease in their children. We conclude that exposure to maternal smoking can increase the variability of PEFR and thus might contribute to the development of asthma. PMID- 8404181 TI - Are asthmatics salt-sensitive? A preliminary controlled study. AB - OBJECTIVE: Epidemiologic evidence suggests that high levels of salt consumption are associated with "spastic" disorders of smooth muscles, ie, essential hypertension and bronchial asthma. Experimentally, it has been shown that high intake of salt leads to increased bronchial hyperreactivity in asthmatics, ie, enhanced contractility of bronchial muscle to spasmogenic stimuli. On the basis of these observations, the following questions were asked: (1) Does salt loading worsen the clinical and functional findings in asthmatics? (2) Is it the sodium or the chloride in salt that is important? METHODS: To answer these questions, the effect of salt restriction (= 5 to 6 g NaCl/d = 86 to 103 mmol Na), salt loading (+ 6.1 +/- 2.8 g NaCl/d = + 105 +/- 48 mmol Na), and loading with sodium citrate in nearly equimolar concentrations (+ 140 +/- 40 ml Shohl's solution, = + 120 +/- 30 mmol Na) was investigated in 14 asthmatics in a controlled crossover study. The total sodium load during the high salt diet was 191 to 209 mmol of sodium per day and during the sodium-citrate phase, 206 to 223 mmol of sodium per day. RESULTS: Statistical analysis showed that salt loading worsened symptoms (p = 0.06) and increased the use of inhaled steroids (p < 0.05). The effect on lung function was less equivocal: salt loading worsened the forced expiratory volume in 1 s (p < 0.01) and the peak expiratory flow rate (p < 0.05). This effect was presumably mediated by sodium, not chloride, as is demonstrated by loading with sodium citrate. CONCLUSION: Patients with bronchial asthma seems to be salt sensitive, the responsible ion being presumably sodium. A low-salt diet appears to have a favorable effect in patients with asthma and to reduce the need for anti-asthma drugs. PMID- 8404182 TI - The Knowledge, Attitude, and Self-Efficacy Asthma Questionnaire. AB - The Knowledge, Attitude, and Self-Efficacy Asthma Questionnaire (KASE-AQ) is a paper-and-pencil instrument that was developed to allow physicians, behavioral scientists, and other health care personnel to assess asthma patients' knowledge regarding asthma, their attitudes about their asthma (including their willingness to cooperate with the physician in managing asthma), and their self-efficacy regarding their perceived ability to control the disorder. The KASE-AQ assesses changes in these patient variables following a particular intervention. The KASE AQ proved to be reliable and internally consistent, and a factor analysis revealed presence of three subscales in the questionnaire (knowledge, attitude, and self-efficacy about asthma). Following asthma education and self-management training, experimental group subjects showed significant improvements in knowledge, attitude, and self-efficacy. Waiting-list control subjects showed similar improvements following training. Both groups' scores at 3-month follow-up remained significantly higher than their baseline scores on all 3 variables. PMID- 8404183 TI - The diagnostic accuracy of high-resolution computed tomography in diffuse infiltrative lung diseases. AB - The purpose of this study was to evaluate the role of high-resolution computed tomography (HRCT) in the clinical diagnosis of diffuse infiltrative lung disease (DILD). Diagnostic accuracy was compared using both chest radiography and HRCT. One hundred thirty-four cases of DILD, representing 21 different diseases, were selected for study, and the disease state was confirmed either histologically or microbiologically. The HRCT images and chest radiographs, available in all cases, were reviewed separately and in random order by 20 physicians who were provided only with information on each patient's age and sex. Overall, a correct first choice diagnosis was made in 38 percent using radiographs and in 46 percent using HRCT images (p < 0.01). The correct diagnosis was among the top three choices in 49 percent when chest radiographs were used, and in 59 percent when HRCT images were viewed (p < 0.01). The correct first-choice diagnosis increased remarkably when the HRCT was used in usual interstitial pneumonia, sarcoidosis, alveolar proteinosis, bronchiolitis obliterans organizing pneumonia, hypersensitivity pneumonitis, and pulmonary lymphangiomyomatosis. High-resolution computed tomography was confirmed to be superior to conventional radiography in the accurate diagnosis of DILD in clinical practice. PMID- 8404184 TI - High-resolution CT in simple coal workers' pneumoconiosis. Lack of correlation with pulmonary function tests and arterial blood gas values. AB - We examined 21 miners by means of standard chest radiography, high-resolution computerized tomography (HRCT), pulmonary function tests, and resting arterial blood gas levels. Using the ILO/UC classification of pneumoconiosis, 7 miners had category 1/0 or 2/1 simple coal workers' pneumoconiosis (CWP). By HRCT, nodules were identified in 12 miners; 4 of 9 were classified as category 0/0 CWP; 2 of 5, 0/1 CWP; 5 of 6, 1/0 CWP; and 1 of 1, 2/1 CWP by chest radiograph. Focal emphysema was identified by HRCT in 7 miners; 4 of 9 were classified as 0/0 CWP; 2 of 5, 0/1 CWP; and 1 of 6, 1/0 CWP by standard chest radiography. Four miners with definite nodules confirmed by HRCT had focal emphysema, while three without nodules had focal emphysema. Pulmonary function testing was not different between miners with or without CWP by standard chest radiography, nor was it different between miners with or without definite nodules evidenced by HRCT. No difference in resting oxygenation was found between any group of miners. The presence of focal emphysema confirmed by HRCT did not significantly affect pulmonary function tests on resting arterial blood gas values. There was, however, a significantly lower FEV1 and mean forced expiratory flow during the middle half of forced vital capacity with lifetime nonsmoking miners. The presence of CWP on chest radiography was significantly correlated with smoking cigarettes but not the years of mining. The presence of nodules on HRCT approached a significant correlation with cigarette smoking, but focal emphysema did not. For detecting evidence of coal dust accumulation in lung parenchyma and identifying focal emphysema, HRCT was more sensitive than standard chest radiography. However, despite earlier detection of parenchymal abnormalities, abnormal pulmonary function attributable to coal dust could not be identified. PMID- 8404185 TI - Laminin fragment P1 is increased in the lower respiratory tract of patients with diffuse interstitial lung diseases. AB - Laminin is a 900,000-dalton extracellular matrix glycoprotein involved in a variety of functions, including cellular movement, growth, and differentiation. The aim of this work was to investigate the presence and biologic significance of this substance in the bronchoalveolar lavage fluid (BALF) of diffuse interstitial lung diseases (DILD). Levels of laminin fragment P1 (LFP) were measured by radioimmunoassay in BALF and sera from controls (n = 8) and patients with several types of DILD: sarcoidosis (n = 10), neoplastic pulmonary infiltration (n = 8), pulmonary fibrosis (n = 5), and hypersensitivity pneumonitis (n = 5). Furthermore, their relation to signs of alveolitis (cellular profiles and albumin concentration in BALF) and evidence of pulmonary fibroblast activation (BALF aminoterminal propeptide of type III procollagen) was examined. Laminin fragment P1 immunoreactivity was detectable in BALF, even in the control group, but patients with all types of DILD had higher concentrations than the control subjects. The serum levels of LFP were similar in all groups studied. Neutrophil and lymphocyte proportions were significantly higher in all DILD groups than in the control group. A positive correlation was seen between lymphocyte proportion and laminin fragment P1 in BALF. Moreover, in BALF a positive correlation was found between LFP and albumin and between LFP and the aminoterminal propeptide of type III procollagen. The BALF macrophage-associated laminin fragment P1 was significantly higher in the active sarcoidosis subgroup compared with the control group. Thus, laminin is a normal constituent of the epithelial lining fluid. The increase of laminin in BALF of patients with DILD suggests that laminin may contribute to their pathogenesis. PMID- 8404186 TI - HLA class I, II, and III polymorphism in Italian patients with sarcoidosis. The Pavia-Padova Sarcoidosis Study Group. AB - We studied the HLA polymorphisms (class I, II, and III) in 107 Italian patients with biopsy specimen-proven sarcoidosis in order to investigate the immunogenetic background of this disease. The mean age of onset of the disease was 36.08 +/- 12.4 years. Four patients (3.73 percent) were in radiologic stage 0, 38 patients (35.51 percent) were in radiologic stage I, 40 patients (37.38 percent) were in stage II, and 25 (23.36 percent) were in stage III. Thirty-eight patients (35.51 percent) had one or more extrapulmonary localization(s) of the disease. Positive association between sarcoidosis and HLA-B8 (chi 2 = 6.07, p = 0.0127, RR = 1.91) was confirmed. Regarding the age of onset of the disease, HLA-B35 was more frequent (chi 2 = 7.34, p = 0.0056, pc < 0.05, RR = 4.62) in patients with early onset of symptoms and/or signs, before the mean age of 36 years. With reference to the radiologic stage of the disease, HLA class II marker DR3 was more frequent in patients with stage I (chi 2 = 7.22, p = 0.0061, pc < 0.05, RR = 7.08). No significant relationship was found between sarcoidosis and HLA class III markers. These results seem to confirm an association of sarcoidosis with HLA classic genes and can sustain the hypothesis of a genetic heterogeneity of this disease. PMID- 8404187 TI - Pulmonary manifestations in lysinuric protein intolerance. AB - STUDY OBJECTIVES: To evaluate the pulmonary manifestations and the course of acute respiratory insufficiency associated with lysinuric protein intolerance (LPI). DESIGN: Retrospective review of clinical data and chest radiographs (total 225) obtained during the lifetime follow-up of 31 LPI patients. About half of the 25 patients without respiratory symptoms underwent high-resolution computed tomography (HRCT) of the lungs, radionuclide perfusion imaging, whole body plethysmography, and diffusing capacity measurements. PATIENTS: Thirty-one Finnish patients with LPI. RESULTS: During the follow-up period, four children with LPI died in respiratory insufficiency, 1 adult had an episode of respiratory insufficiency, and another had chronic symptoms, whereas 25 patients remained symptom-free. The radiologic findings in acute progressive respiratory insufficiency were uniform: at first, reticulonodular interstitial densities and, later on, progressive airspace disease. At autopsy, three patients showed pulmonary alveolar proteinosis and one had pulmonary hemorrhage and cholesterol granulomas. One adult had reversible respiratory insufficiency with signs of bronchiolitis obliterans, another adult had recurrent episodes of chest pain, dyspnea, and hypoxia. Of the symptom-free patients, one third (8 of 25) had signs suggestive of pulmonary fibrosis evidenced on chest radiographs and two thirds (8 of 14) had signs evidenced by HRCT films. Most symptom-free patients showed mild abnormalities either in perfusion imaging (9 of 12) or in function tests (8 of 12). CONCLUSION: In childhood, patients with LPI are highly predisposed to develop pulmonary hemorrhages and alveolar proteinosis. Interstitial lung densities may precede the acute phase. Most adult LPI patients show radiologic signs of interstitial lung disease but only a few show clinical impairment. PMID- 8404188 TI - Follicular bronchitis in the pediatric population. AB - Five patients in a pediatric population were identified with idiopathic follicular bronchitis (IFB) by open lung biopsy and their case records were reviewed. All were tachypneic and had a chronic cough by 6 weeks of age. The physical examination was characterized by diffuse fine crackles in four patients and by coarse rhonchi in one. The chest radiographs in all demonstrated a diffuse interstitial pattern. None had a collagen vascular or an autoimmune disease demonstrable. Response to corticosteroid therapy was minimal. Associated or coincidental esophageal reflux was treated surgically in two. No viral or bacterial agents were isolated in the sputum or the biopsy specimens. Patients have been followed up for 2 to 15 years; the conditions of all patients improved at about 2 to 4 years of age. The older patients have residual mild obstructive lung disease. To our knowledge, this is the first reported series of IFB in the pediatric population. PMID- 8404189 TI - Accuracy of three pulse oximeters during exercise and hypoxemia in patients with cystic fibrosis. AB - To evaluate the reliability of noninvasive oximeters during exercise in patients with cystic fibrosis (CF), we studied 9 patients during exercise tests, using 3 devices (Hewlett-Packard 47201A, Biox-Ohmeda 3700, Nellcor 100), comparing oximetry readings with arterial blood co-oximetry. Fifty samples from preexercise, exercise, and recovery were collected; and paired measurements were analyzed for bias (arterial blood gas SaO2--noninvasive SaO2) and precision (1 SD of the bias). Values were available for the Hewlett-Packard and the Biox-Ohmeda at all 50 collection times. The Nellcor failed to read on four occasions, all during exercise. The accuracy of the Biox-Ohmeda readings was significantly worse during exercise than rest, while the Nellcor and the Hewlett-Packard performance did not differ significantly between exercise and rest. During hypoxemia (SaO2 < 90 percent as determined by co-oximetry on arterial blood), the Biox-Ohmeda and Nellcor consistently overestimated saturation, with mean bias of -6.1 percent and -3.0 percent, respectively. The Hewlett-Packard was more reliable and valid during hypoxemia, with a mean bias of -1.0 percent. The Nellcor read significantly more accurately in the six patients with severe digital clubbing than in the three patients with mild digital clubbing. It is important to be aware of possible limitations of noninvasive oximeters before accepting them as accurate. PMID- 8404190 TI - Anti-inflammatory action of erythromycin. Its inhibitory effect on neutrophil NADPH oxidase activity. AB - The effects of erythromycin on the NADPH oxidase activity in the neutrophils of normal subjects and patients with bronchiolitis were investigated. In the patients receiving erythromycin, NADPH oxidase activity was significantly lower in a whole-cell system than that before therapy. Erythromycin was also found to inhibit the superoxide generation of neutrophils exposed to phorbol myristate acetate in a whole-cell system and the activation of superoxide-generating NADPH oxidase by sodium lauryl sulfate (sodium dodecyl sulfate) in a cell-free system. The concentration of the drug required for 50 percent inhibition of the oxidase was 0.7 mM in the whole-cell system and 0.2 mM in the cell-free system. These results suggest that erythromycin, an antibiotic which penetrates well into human neutrophils, may exhibit an anti-inflammatory action due to inhibiting of neutrophil NADPH oxidase activation. PMID- 8404191 TI - Controlled trial of ciprofloxacin in short-term chemotherapy for pulmonary tuberculosis. AB - The study was undertaken in patients with newly diagnosed bacteriologically positive pulmonary tuberculosis. The patients were randomly allocated to receive one of two regimens. In one of the regimens, patients received 2 months of daily treatment with streptomycin, isoniazid, rifampin, and pyrazinamide (SHRZ) followed by isoniazid plus rifampin for 4 months (4 HR). In the other regimen, patients received 2 months of daily streptomycin, isoniazid, pyrazinamide, and ciprofloxacin (SHZ Cipro) followed by isoniazid plus ciprofloxacin for 4 months. All patients in the SHRZ/HR regimens and all but one in the SHZ Cipro/H Cipro regimens had a favorable bacteriologic response during chemotherapy. In the patients in the SHZ Cipro/H Cipro regimen, ciprofloxacin was well tolerated during the period of treatment. After chemotherapy, one (5.9 percent) of 17 patients in the SHRZ/HR group and three (16.6 percent) of 18 patients in the SHZ Cipro/H Cipro group relapsed bacteriologically. PMID- 8404192 TI - The effects of transtracheal gas delivery on central inspiratory neuromuscular drive. AB - Previous studies have shown transtracheal delivery of low-flow oxygen (TTO) decreases inspired minute ventilation (Veinsp) and have postulated that this would result in a decrease in the work of breathing (WOB). We hypothesized that a fall in central inspiratory neuromuscular drive (CIND) with TTO would reflect a fall in WOB. We measured resting ventilatory parameters (RVP) and CIND by the mouth occlusion pressure technique (MOP) at different gas flow rates through the catheter in 21 subjects (13 men, 8 women; mean age, 60 +/- 10.6 years) with severe COPD with a mature intratracheal oxygen catheter (ITOC). We also constructed a lung/chest wall analog (LCA) to determine if flow through the catheter would alter pressure changes during inspiration. Inspiratory tidal volume (Vtinsp) and minute ventilation (Veinsp) decreased proportionally to the gas flow rate through the catheter. However, with increasing flow through the catheter, P0.1 increased in the LCA, presumably due to the Bernoulli effect. The lack of a similar change in the subject group suggests that CIND does, in fact, fall, and that possibly there is a decrease in WOB. This effect may be of benefit to patients with severe COPD. PMID- 8404193 TI - Maximum respiratory pressures in trumpet players. AB - We studied whether experienced trumpet players can develop higher pressures with their inspiratory and expiratory muscles than untrained subjects. Twelve male trumpet players (mean age, 22.4 +/- 3.3 years) participated in the study. All of them had played the trumpet for at least 4 years and were nonsmokers. Twelve healthy male subjects (mean age, 23.3 +/- 3.1 years) participated as a control group. There were no differences in spirometric parameters between both groups. Maximum respiratory pressures were higher in the trumpet player group (trumpet players: Pmax 151.3 +/- 19.8 cm H2O; Pemax, 234.6 +/- 53.9 cm H2O; control group: Pemax, 106.7 +/- 10.4 cm H2O; Pemax, 189.6 +/- 14.6 cm H2O). We concluded that in young trumpet players, maximum respiratory pressures are higher than in young people who do not play wind instruments. This is most probably a consequence of respiratory muscle training with a wind instrument. PMID- 8404194 TI - Current management of esophageal impactions. AB - We analyzed our experience at a university medical center from 1977 to 1990 to assess our success in using esophagoscopy and related treatments for removing esophageal impactions. There were 157 episodes of impaction in 150 patients, consisting of 39 pediatric and 111 adult patients. In the pediatric cases, foreign bodies were most often the cause of impaction, while adult cases were usually caused by food or bones. Esophagoscopy was performed successfully in 32 of 34 pediatric patients in which it was attempted; there was only one complication. Other forms of therapy that were infrequently tried met with variable results. Esophagoscopy was successful in removing the impaction in 104 of 109 attempts in adults. Two perforations occurred, with one resulting in death. Various other methods achieved success in the remaining patients. The data suggest that esophageal impaction can be treated successfully by endoscopy with very low morbidity and mortality. PMID- 8404195 TI - Prediction of respiratory symptoms by peripheral blood neutrophils and eosinophils in the First National Nutrition Examination Survey (NHANES I). AB - To examine specific cellular markers of inflammation in peripheral blood (neutrophils and eosinophils) and their relationship to respiratory symptoms, we used data from the First National Health and Nutrition Examination Survey (NHANES I). Cross-sectional data were available on a random sample of 6,913 adults aged 30 to 74 years who had American Thoracic Society-National Heart, Lung, and Blood Institute questionnaire information on respiratory symptoms and illnesses, including asthma, chronic bronchitis, dyspnea (grade 3), chronic cough, and chronic phlegm. Information was available on age, race, smoking status, peripheral blood leukocyte count, and differential cell count. These data were analyzed using logistic regression controlling for age, race, sex, and cigarette smoking. Physician-diagnosed asthma was significantly associated only with the eosinophil count (p = 0.001). Physician-diagnosed bronchitis was significantly associated with the neutrophil count (p = 0.012) and marginally associated with the eosinophil count (p = 0.072). Chronic phlegm was also significantly associated with both the eosinophil count (p = 0.049) and the neutrophil count (p = 0.041). Grade 3 dyspnea (p = 0.049) was only significantly associated with the neutrophil count. These data suggest that both peripheral blood neutrophils and eosinophils are associated with a broad range of respiratory symptoms and that the eosinophil may play a role in nonasthmatic respiratory inflammation. PMID- 8404196 TI - High-frequency ventilation for acute pediatric respiratory failure. AB - OBJECTIVE: To study the effectiveness of high-frequency ventilation (HFV) for the treatment of pediatric patients with acute severe respiratory failure. DESIGN: Post hoc analysis of retrospectively and prospectively acquired data. SETTING: Tertiary pediatric ICU. SUBJECTS: Twelve patients, ages 4 months to 15 years, who developed acute severe respiratory failure from diverse causes. INTERVENTIONS: Patients were treated with synchronized intermittent mandatory ventilation (SIMV) using moderate positive end-expiratory pressure (PEEP) and supplemental oxygen prior to HFV therapy. Ten of 12 patients required one or more medications to modulate preload, inotropy, and/or afterload. All patients were invasively monitored for arterial BP and arterial blood gases, and they were noninvasively monitored for oxygen saturation, end tidal or transcutaneous carbon dioxide, and electrocardiography. MEASUREMENTS AND MAIN RESULTS: Severity of respiratory failure was reflected by median values of pH of 7.34, lung compliance of 0.41 ml/cm H2O/kg, P(A-a)O2 of 553 mm Hg, oxygenation index (OI) of 28, and ventilation index of 102. Significant improvement in pH, PaCO2, PaO2, OI, and P(A a)O2 was demonstrated early in the course of HFV (p < 0.05). Seven of the patients (58 percent) were survivors. CONCLUSIONS: These data suggest that HFV may offer significant benefit as a rescue modality for patients with severe respiratory failure refractory to SIMV strategies early in the course of the disease process. PMID- 8404197 TI - Survival following mechanical ventilation for acute respiratory failure in adult men. AB - STUDY DESIGN: Survival following mechanical ventilation for acute respiratory failure has important implications for medical decision-making and allocation of expensive resources for critical care. PROCEDURE: We reviewed a 5-year experience with mechanical ventilation in 383 men with acute respiratory failure and studied the impact of patient age, cause of acute respiratory failure, and duration of mechanical ventilation on survival. Survival rates were 66.6 percent to weaning, 61.1 percent to ICU discharge, 49.6 percent to hospital discharge, and 30.1 percent to 1 year after hospital discharge. When our data were combined with 10 previously reported series, mean survival rates were calculated to be 62 percent to ventilator weaning, 46 percent to ICU discharge, 43 percent to hospital discharge, and 30 percent to 1 year after discharge. Of 255 patients weaned from mechanical ventilation, 44 (17.3 percent) required an additional period of mechanical ventilation during the same hospitalization. RESULTS: Age had a significant influence on survival to hospital discharge and on that to 1 year after hospital discharge, and the cause of acute respiratory failure had a significant influence on survival only to weaning. Survival was best in younger patients and those with COPD or postoperative respiratory failure and worst in patients resuscitated after cardiac or respiratory arrest. Increased duration of mechanical ventilation significantly reduced survival only to hospital discharge. Overall survival was significantly affected by age and cause of acute respiratory failure, but not by duration of mechanical ventilation. CONCLUSION: We conclude that age, cause of acute respiratory failure, and duration of mechanical ventilation have specific influences on the generally poor outcome of mechanical ventilation for acute respiratory failure. PMID- 8404198 TI - Impact of previous antimicrobial therapy on the etiology and outcome of ventilator-associated pneumonia. AB - OBJECTIVE: To define the influence of prior antibiotic use on the etiology and mortality of ventilator-associated pneumonia (VAP). SETTING: A university hospital medical-surgical ICU. DESIGN: Prospective clinical study. METHODS: Over a 35-month period, we prospectively studied 129 consecutive episodes of VAP. Etiologic diagnosis was established using a protected specimen brush and quantitative culture techniques. We examined prognostic factors by univariate and multivariate analyses using a statistical software package (SPSS). RESULTS: The rate of VAP caused by Gram-positive cocci or Haemophilus influenzae was statistically lower (p < 0.05) in the patients who had received antibiotics previously, while the rate of VAP caused by Pseudomonas aeruginosa was statistically higher (p < 0.01). Patients died of causes directly related to the infection in 18 (14.0 percent) episodes, P aeruginosa being isolated in 9 of these fatal cases. Indeed, we found that 27.7 percent (15/54) of patients who had received prior antimicrobial therapy before the onset of pneumonia died, compared with only 4.0 percent (3/75) of those who did not. In the univariate analysis, the variables significantly associated with attributable mortality were age older than 45 years, use of corticosteroids, presence of shock, hospital day of VAP over 9, antecedent COPD, and a prior antibiotic use. A step-forward logistic regression analysis defined only prior antibiotic use (p < 0.0001, OR = 9.2) as significantly influencing the risk of death from VAP. The same result was obtained when severity was included in the model. However, prior antibiotic use entirely dropped out as a significant risk factor when the etiologic agent was included in the regression equation. CONCLUSIONS: Distribution of infecting microorganisms responsible for VAP differs in patients who received prior antimicrobial therapy, and this factor determines a higher mortality rate. We suggest a restrictive antibiotic policy in mechanically ventilated patients with the purpose of reducing the risk of death from VAP. PMID- 8404199 TI - Relationship of thermodilution cardiac output to metabolic measurements and mixed venous oxygen saturation. AB - To determine the individual contributions of variables in the Fick equation to cardiac output, we simultaneously measured oxygen uptake (VO2), carbon dioxide production (VCO2), venous oxygen saturation (SvO2) and thermodilution cardiac output (Qth) in 28 medical and surgical ICU patients. Patients were intubated and ventilated with the intermittent mandatory ventilation mode. VO2 and VCO2 (averaged over 3 min) were obtained from a metabolic cart. SvO2 was measured with fiberoptic reflectance oximetry (and COoximetry). Thirty-nine studies (average duration, 4.3 h) with 151 Qth measurements were performed. The relationships between Qth and VO2, Qth and VCO2, Qth and SvO2, and 1/Qth and SvO2, as well as between the sequential changes in these variables were analyzed by least squares linear regression. The ability of changes in the variables VO2, VCO2, and SvO2 to predict changes in Qth were analyzed by receiver operating characteristic (ROC) curves. Qth was weakly related to VO2 (r = 0.45), VCO2 (r = 0.45), or SvO2 (r = 0.36). Changes in Qth were weakly related to changes in VCO2 (r = 0.40), and even less to changes in VO2 (r = 0.18) and SvO2 (r = 0.13). The areas under the ROC curves for increases in Qth > 10 percent were as follows: 0.66 for VCO2, 0.50 for VO2, and 0.55 for SvO2. The areas for decreases in Qth < 10 percent were as follows: 0.78 for VCO2, 0.65 for VO2, and 0.49 for SvO2. None of the above oximetry relationships were substantially altered by use of COoximetry venous oxygen saturations. We conclude that Qth cannot be predicted well solely from VO2, VCO2, or SvO2 nor can changes in Qth be predicted well solely from changes in VO2, VCO2, or SvO2. Of the metabolic variables, changes in VCO2 best predicted changes in Qth. PMID- 8404200 TI - Thrombocytopenia in the intensive care unit. AB - STUDY OBJECTIVE: To determine the prevalence of thrombocytopenia in an ICU and assess which factors were associated with thrombocytopenia. DESIGN: A review of the medical records of patients admitted during 3 separate months during 1 academic year. Patients must have survived at least 12 h in the ICU. SETTING: A medical ICU at a university hospital. PATIENTS: General medicine patients admitted to the ICU. INTERVENTIONS: All medical records were reviewed. During the ICU stay, daily medications, events, and platelet count were noted. All patients were followed up until death or hospital discharge. In 22 patients, including 18 who had thrombocytopenia, bone marrow aspirates were performed. MEASUREMENTS AND RESULTS: One hundred sixty-two admissions were evaluated. Thirty-eight (23 percent) had platelet counts less than 100,000/mm3 at least once, and 17 (10 percent) patients had platelet counts less than 50,000/mm3. Several factors were associated with thrombocytopenia; however, only sepsis, use of antineoplastic chemotherapy, elevated creatinine level, or elevated bilirubin value were independent risk factors for severe thrombocytopenia. In only one patient were the bone marrow findings different from those expected by the clinical presentation. Thrombocytopenia was associated with longer hospital stay (p < 0.001) and higher mortality (p < 0.001). CONCLUSION: Thrombocytopenia is a common occurrence in the ICU, usually due to the underlying disease, and is associated with an increased mortality. PMID- 8404201 TI - Variables predictive of outcome in 832 patients undergoing repairs of the descending thoracic aorta. PMID- 8404202 TI - Gastric intramural PCO2 during peritonitis and shock. AB - OBJECTIVE: To define whether increases in gastric intramural tissue CO2 and H+ increase during experimentally induced peritonitis with circulatory shock as they do under conditions of hemorrhagic shock and cardiac arrest. DESIGN AND SETTING: Peritonitis was induced in Sprague-Dawley rats by cecal ligation and fecal spillage. MEASUREMENTS AND RESULTS: Over an interval of 260 +/- 20 min in 5 animals, there was a progressive reduction in mean aortic pressure from 153 +/- 12 to 40 +/- 20 mm Hg and a decline in cardiac index from 429 +/- 135 to 178 +/- 7 ml/min. This was associated with increases in gastric intramural [H+] from 34 +/- 5 to 217 +/- 93 mmol/L (p = 0.001). Arterial blood lactate content concurrently increased from 0.9 +/- 0.1 to 4.6 +/- 0.7 mmol/L (p = 0.001). Only a late increase in gastric intramural PCO2 from 45 +/- 5 to 128 +/- 38 mm Hg (p = 0.01) was observed. CONCLUSION: In contrast to the gastric acid base changes that accompany hemorrhagic shock, in which there is an early and prominent increase in both PCO2 and [H+] in close relationship to decreases in cardiac output and arterial pressure, there was a prominent increase in gastric [H+] but only a delayed rise in gastric intramural PCO2. Arterial blood lactate and central venous oxygen saturation were earlier indicators of perfusion failure. Since the bicarbonate concentration in the stomach wall was substantially greater than that of simultaneously measured arterial blood, this has bearing on the current clinical method of gastric tonometry which assumes that arterial blood bicarbonate is equivalent to gastric wall bicarbonate. PMID- 8404203 TI - Video-assisted thoracic surgical techniques in the diagnosis and management of pericardial effusion in patients with advanced lung cancer. PMID- 8404204 TI - Dyspnea of 2-weeks' duration and left pleural effusion in a previously healthy 50 year-old woman. PMID- 8404205 TI - Cheerios in the chest. PMID- 8404206 TI - Thoracoscopic diagnosis of pleurolithiasis after laparoscopic cholecystectomy. AB - We describe a patient with right pleuritic chest pain and an enlarging exudative pleural effusion four months after laparoscopic cholecystectomy. Several radiographic imaging procedures and thoracenteses were nondiagnostic. Thoracoscopy, however, revealed bilious concretions in the parietal pleura. Thoracoscopic drainage, lysis of adhesions, and antibiotic treatment of a Klebsiella pneumoniae pleuritis resulted in relief of symptoms. PMID- 8404207 TI - Tetracycline pleurodesis--associated acute renal failure. AB - Chemical pleurodesis is a frequently performed procedure for pneumothorax and effusion and significant adverse effects are unusual. We present a previously unreported case of acute renal failure associated with tetracycline pleurodesis. Recent studies have shown that intrapleural drug administration may lead to therapeutic serum levels. Systemic toxic drug effects may therefore be noted with chemical pleurodesants such as tetracycline. Alternative methods of pleurodesis should always be considered if a sensitivity or metabolic abnormality is suspected. PMID- 8404208 TI - Bilateral pleural effusion due to mediastinal fibrosis induced by radiotherapy. AB - A diagnosis of Hodgkin's disease was made 20 years ago in a 37-year-old woman; treatment included thoracic irradiation and chemotherapy. She was considered cured and remained well up to one year before, when she developed bilateral pleural effusion. No evidence of activity of Hodgkin's disease was detected. The pleural liquid was an exudate, with lymphocytic predominance. On thoracoscopy, enlarged lymphatic channels in the visceral pleura were noted, with tissue confirmation. To our knowledge, this report is the first to confirm by thoracoscopy and histologic study the proposed pathophysiologic condition of this uncommon entity. PMID- 8404209 TI - Metastatic carcinoma of the prostate presenting as a superior vena cava syndrome. AB - A 75-year-old patient presented with a superior vena cava syndrome (SVCS) lasting 3 years. A prostatic carcinoma was found and a supraclavicular lymph node biopsy specimen disclosed metastasis of the prostatic carcinoma. Antiandrogen and luteinizing hormone-releasing hormone analogue therapy produced a marked improvement. Prostatic carcinoma, although a very rare cause, must be considered in the diagnosis of cases of SVCS with a protracted course, since it is a treatable disease. PMID- 8404210 TI - Detection of intrapulmonary shunts in schistosomal cor pulmonale. AB - Two patients with schistosomal cor pulmonale and central cyanosis were studied by contrast-enhanced echocardiography, using indocyanine green injection. Intrapulmonary shunts were detected by this method. To our knowledge, this is the first report that proves the presence of intrapulmonary shunts in schistosomal cor pulmonale detected by contrast-enhanced echocardiography. PMID- 8404211 TI - Radiation-induced pulmonary veno-occlusive disease. AB - Late occurrence of radiation-induced pulmonary pneumonitis and fibrosis is well documented. We report an unusual case of radiation induced veno-occlusive disease (VOD) occurring six years following mantle irradiation for Hodgkin's lymphoma. The patient developed severe pulmonary hypertension and cor pulmonale. A left lung transplantation was performed successfully and pathologic examination of the explanted lung showed severe changes compatible with VOD. In the absence of exposure to alternate therapeutic or toxic agents that may cause VOD, it is likely that radiation caused damage to the venular endothelium and caused progressive obliteration of the pulmonary vessels. Review of the literature reveals only a few similar reports of VOD mostly following radiation for bone marrow transplantation. We conclude that previous irradiation (even several years earlier) should be considered as a possible cause of pulmonary VOD. PMID- 8404212 TI - Confirmation of anomalous origin of the right coronary artery from the left sinus of Valsalva with magnetic resonance imaging. AB - Anomalous origin of the right coronary artery from the left sinus of Valsalva is a rare but clinically significant congenital abnormality, difficult to diagnose angiographically. We describe a patient in whom magnetic resonance imaging was used to delineate the anomalous course of the right coronary artery following angiographic demonstration limited by technical considerations. PMID- 8404213 TI - Bronchiectasis in a child after acrolein inhalation. AB - Acrolein is an unsaturated aldehyde produced by combustion of many organic compounds. Massive exposure may lead to severe pulmonary disease and possibly death. We report a case of intoxication in a 2-year-old boy; an 18-month follow up showed development of obstructive bronchiolar disease with diffuse bronchiectasis. PMID- 8404214 TI - Cheyne-stokes ventilation converting to obstructive sleep apnea following heart transplantation. AB - Cheyne-Stokes ventilation is often found in conjunction with heart failure. The pathogenesis is multifactorial and upper airway instability has been suggested to play a role. This report documents the conversion of Cheyne-Stokes ventilation during sleep to obstructive apnea after heart transplantation. PMID- 8404215 TI - Wegener's granulomatosis with peripheral eosinophilia. Atypical variant of a classic disease. AB - A patient with Wegener's granulomatosis (WG) diagnosed by ultrasound-guided transthoracic biopsy of a pulmonary nodule is reported. The case is atypical because of marked eosinophilia in the peripheral blood and the pleural effusion. The granulomatous infiltrate of the lung showed the classic picture of WG without eosinophils. The patient responded dramatically to treatment with steroids and cyclophosphamide. This variant form of WG poses problems in its distinction from Churg-Strauss syndrome, and the differential diagnosis between these two entities is discussed. PMID- 8404216 TI - Primary extranodal non-Hodgkin's lymphoma of the lung presenting with bilateral, patchy infiltrates dramatically improving after corticosteroid therapy. AB - A 63-year-old man was admitted to the hospital with fever and bilateral, peripheral infiltrates. Infectious disease and malignancy seemed to be excluded by fiberoptic diagnostic procedures. Subsequently, respiratory insufficiency developed, making open lung biopsy impossible. The diagnosis of bronchiolitis obliterans organizing pneumonia (BOOP) was strongly considered and treatment with corticosteroids was started; this led to dramatic clinical and radiologic improvement for a short time. Eventually, an open lung biopsy specimen disclosed primary extranodal non-Hodgkin's lymphoma of T-cell origin, immunoblastic, of high-grade malignancy according to the Kiel classification. After the first course of chemotherapy, total respiratory insufficiency developed and the patient died. This case is unique in a patient without AIDS. PMID- 8404217 TI - Bronchiolitis obliterans organizing pneumonia caused by Plasmodium vivax malaria. AB - A 64-year-old woman developed a relapse of Plasmodium vivax malaria followed by a rapidly progressive diffuse patchy pulmonary process. Open lung biopsy specimen showed bronchiolitis obliterans organizing pneumonia (BOOP). After corticosteroid therapy was initiated, there was both clinical and radiographic improvement. This is believed to be the first reported association of BOOP with malaria. PMID- 8404218 TI - Sepsis syndrome and death after bronchoalveolar lavage. AB - Bronchoalveolar lavage is widely used in the management of patients with interstitial lung diseases and is considered a safe procedure. We describe a patient who died with a picture consistent with acute pulmonary edema and septic shock following bronchoalveolar lavage. This potential complication has not been previously reported. PMID- 8404219 TI - Interstitial lung disease and myositis in a patient with simultaneously occurring sarcoidosis and scleroderma. AB - A patient initially presented with sarcoidosis in combination with myositis of sarcoid origin and Raynaud's phenomenon. During the course of his disease, he additionally developed scleroderma. Bronchoalveolar lavage, performed because of increase of interstitial markings in the presence of enlarged hilar nodes, showed an increased percentage of granulocytes and a low CD4 to CD8 ratio of lymphocytes, suggesting a change to scleroderma lung disease. PMID- 8404220 TI - Bronchospasm secondary to replacement estrogen therapy. AB - A postmenopausal woman with severe obstructive airways disease and bronchospasm developed increased airflow limitation with the reintroduction of estrogen therapy for osteoporosis. Discontinuation of the estrogen caused symptomatic improvement and decreased her corticosteroid requirement. Readministration of estrogen caused recrudescence of her symptoms and a decline in her peak expiratory flow rate and spirometric data, which reversed with withdrawal of the estrogen therapy. Bronchospasm during the luteal phase of the menstrual cycle is well known, but exacerbation of reactive airways disease with the administration of exogenous estrogen has not previously been reported; however, with the increasing practice of reintroducing estrogen in postmenopausal women to reduce the risk of symptomatic osteoporosis, other susceptible women may suffer clinically significant deterioration of their underlying pulmonary disease. PMID- 8404221 TI - The flow-volume loop in bilateral vocal cord paralysis. AB - A 38-year-old man with posttraumatic bilateral vocal cord paralysis and a surgically repaired avulsion of the extrathoracic trachea presented with a slight increase of exertional dyspnea (grade 2). Spirometry showed high normal FEV1 for FVC variables, but the F-V loop was characteristic for highly variable UAO with an increased FEV1/PEF ratio of 11 ml/L/min as well as a MEF50/MIF50 of 4.55. Endoscopy during forced respiration showed near total inspiratory obstruction of the larynx due to paradoxical behavior of the vocal cords. In extrathoracic airway obstruction a FEV1/PEF ratio > 10 ml/L/min combined with a MEF50/MIF50 ratio > 4 is suggestive of variable UAO caused by bilateral vocal cord paralysis rather than by a tracheal lesion. PMID- 8404222 TI - Severe extrapulmonary thoracic restriction caused by morphea, a form of localized scleroderma. AB - A prevalent clinical perception is that thoracic restriction in patients with morphea or scleroderma should not result from cutaneous sclerosis alone; that there must be some underlying parenchymal lung disease or respiratory muscle weakness. But herein we describe a patient with morphea and severe thoracic restriction that appears to result mainly from cutaneous sclerosis. PMID- 8404223 TI - Platypnea in the intensive care unit. A newly described cause. AB - Platypnea in a patient with COPD developed during the subacute onset of an ileus. Arterial blood gas studies failed to document orthodeoxia. Routine treatment for COPD failed to resolve the patient's positional dyspnea, but the dyspnea rapidly resolved following resolution of the ileus. The authors postulate that impaired abdominal muscle contraction in the upright position secondary to the ileus was responsible for the development of platypnea. PMID- 8404224 TI - Major hemorrhage as a complication of cough fracture. PMID- 8404225 TI - Upper airway obstruction following angiotensin-converting enzyme inhibitor therapy. PMID- 8404226 TI - Critical care board examination resources. PMID- 8404227 TI - Role of upper airway occlusion as a predisposing factor for systemic hypertension. PMID- 8404228 TI - Postprandial serum cholylglycine as a marker of antituberculous drug hepatotoxicity in HIV-infected patients. PMID- 8404229 TI - Endotracheal tube cuff perforation as a complication of subclavian vein catheterization. PMID- 8404230 TI - Pulmonary arteritis with pulmonary arterial thrombosis and recurrent endopulmonary embolization. PMID- 8404231 TI - Pleural determinants of restrictive lung function and respiratory symptoms in an asbestos-exposed population. PMID- 8404232 TI - Nasal mask ventilation in acute respiratory failure. PMID- 8404233 TI - Nasal mask ventilation in acute respiratory failure. PMID- 8404234 TI - Beware of the VIP syndrome. PMID- 8404235 TI - Environmental tobacco smoke and asthma. PMID- 8404236 TI - The nose and obstructive sleep apnea. PMID- 8404237 TI - Mortality and ventilator-associated pneumonia. The best antibiotics may be the least antibiotics. PMID- 8404238 TI - Trending of oxygen utilization in the critically ill. Is there a reliable method? PMID- 8404239 TI - Diagnostic efficacy of PET-FDG imaging in solitary pulmonary nodules. Potential role in evaluation and management. AB - BACKGROUND: Positron emission tomography (PET), a new noninvasive imaging modality, utilizing 2-[F-18]-fluoro-2-deoxy-D-glucose (FDG), has demonstrated increased FDG uptake in lung tumors. OBJECTIVE: To determine the diagnostic efficacy of PET-FDG imaging in differentiating benign from malignant solitary pulmonary nodules. PATIENT SELECTION: A prospective study of 30 patients who presented with indeterminate solitary pulmonary nodules less than 3 cm in size based on chest radiograph and computed tomographic (CT) scan. SETTING: Two tertiary care medical centers in Omaha, Neb: Creighton University Medical Center and the Omaha Veterans Administration Medical Center. MEASUREMENTS: Positron emission tomographic imaging of the lung was performed 1 h after intravenous injection of 10 mCi of F-18-FDG. Qualitative analysis of the images was performed independently by two observers by visual identification of the areas of increased FDG uptake in the lung nodules. Semiquantitative analysis was performed using computation of differential uptake ratio (DUR). Histologic specimens were obtained in 29 patients (thoracotomy 20, transthoracic needle aspiration biopsy 8, bronchoscopy 1). RESULTS: Positron emission tomographic imaging correctly identified 27 of 30 pulmonary nodules. Diagnostic accuracy was high with sensitivity of 95 percent and specificity of 80 percent. The positive and negative predictive value of PET imaging for solitary pulmonary nodules was 90 percent and 89 percent, respectively. The DUR values were significantly higher for malignant nodules (mean +/- SD, 5.55 +/- 2.79) than benign nodules (mean +/- SD, 0.95 +/- 0.99) (p < 0.001). There was one false-negative result in a patient with a 1-cm nodule identified as a scar adenocarcinoma. There were two false positive cases and both had caseating granulomas with active inflammation and Histoplasma organisms. CONCLUSION: PET-FDG imaging of the lung, a new noninvasive diagnostic test, has a high degree of accuracy in differentiating benign from malignant pulmonary nodules. PET-FDG imaging could complement CT scanning in the evaluation and treatment of patients with solitary pulmonary nodules. PMID- 8404240 TI - Parental abuse as a precursor to childhood onset depression and suicidality. AB - Child guidance clinic cases were reviewed for abuse history, diagnosis, depressive symptoms, and suicidality. Many depressed abused children were found to have been previously diagnosed exclusively with conduct-related versus affect disorders. The results suggested that abused children are at specific risk of developing depression and suicidality and that a subgroup of those with early onset depression may have acquired the disorder concurrent with parental abuse. PMID- 8404241 TI - Drug and placebo side effects in methylphenidate-placebo trial for attention deficit hyperactivity disorder. AB - In a double-blind methylphenidate placebo trial for children with Attention Deficit Hyperactivity Disorder (ADHD), side effects were reported on both placebo and methylphenidate (MPH), and many side effects were similar to the symptoms of ADHD. We suggest that parents and teachers may mistake the symptoms of ADHD for side effects of MPH, and this misconstrual may contribute to the poor acceptance of and compliance with taking MPH. PMID- 8404242 TI - Suicide in the psychiatric hospital. AB - Society's attitudes toward and treatment of the suicider are examined. The emotional and clinical impact suicide has on everyone, and its consequences to the hospital are detailed. Our unfortunate emphasis on "prevention" as against treating the disease process which causes the death is stressed. Also stressed is the psychiatrist's misplaced role in contrast to that of other physicians involved in a patient's death. Only in psychiatry does it seem that the role assigned the physician is that the he prevent a specific patient's death as much as that he treat the patient's underlying fatal disease. PMID- 8404243 TI - Residential versus day treatment for children: a long-term follow-up study. AB - Adults and adolescents, who as children, received day or residential treatment were followed up ten years after treatment completion. No differences were found between the two treatment modalities on follow-up ratings of personal and social adjustment. This pattern was not influenced by gender, IQ, or presenting problem. Overall, about two-thirds of the children demonstrated improvement at follow-up. As expected, individuals demonstrating higher levels of personal and social adjustment at initial assessment were functioning better at outcome. However, the magnitude of therapeutic gain was not significantly different for mild versus moderate-to-severe presenting problems. Suggestions for future research are included. PMID- 8404244 TI - Alcoholism prevention among Native-American youth. AB - Although Native-Americans represent a diverse population, alcoholism prevention programs need to apply general knowledge of alcohol use and misuse, rather than search for extraordinary cultural factors. Such an approach would emphasize the importance of social relationships, peer group associations, family interactions, and individual adjustment in the prevention of alcoholism among Native-Americans. PMID- 8404245 TI - Perceived parental characteristics of patients with obsessive compulsive disorder, depression, and panic disorder. AB - It has been hypothesized that parents of patients with obsessive compulsive disorder exhibit specific traits. 320 consecutive inpatient admissions who met criteria for OCD, depression, and panic disorder checked a list of adjectives to describe their parents. Patients with OCD were 1) less likely to perceive their mothers as disorganized than depressives, 2) more likely to perceive their mothers as overprotective than depressives and 3) less likely to perceive their fathers as demanding than patients with panic. PMID- 8404246 TI - Anxiety disorders and pediatric continuous ambulatory peritoneal dialysis. AB - This paper compared 23 children with end-stage renal disease (ESRD) on continuous ambulatory peritoneal dialysis to a matched sample of healthy children. The rate of separation anxiety disorder was significantly higher in the children with ESRD. In their family environments, mothers of ESRD children reported significantly lower independence and achievement orientation than mothers of control children. PMID- 8404247 TI - Child welfare services for the catastrophically ill newborn: Part II--A guiding ethical paradigm. PMID- 8404248 TI - Decision-making in child protective services: a comparison of selected risk assessment systems. AB - The use of structured risk-assessment systems by child protective services has increased dramatically during the past ten years. Versions of the Illinois CANTS 17B, the Washington Assessment of Risk Matrix, and the Child at Risk Field System have been put into use in some form in 23 states [Berkowitz 1991]. Despite this widespread acceptance, workload pressures, uncertain reliability and validity of the instruments, and the need for a knowledgeable and well-trained casework staff have limited the models' usefulness once implemented. PMID- 8404249 TI - A Spanish translation of the Revised Behavior Problem Checklist. AB - As part of a study on the mental health needs of Hispanic children, behavioral assessments were obtained using the Revised Behavior Problem Checklist (RBPC). Although a Spanish translation was available, it had been designed for a primarily Cuban population. Because the population under study was approximately 80% Puerto Rican, 15% Mexican, and 5% Dominican Republican, a new Spanish version was developed. It is suggested that future researchers be sensitive to the differences in vocabulary among Hispanic subcultures. PMID- 8404250 TI - The quality of regulated family day care homes and compliance with minimum standards. AB - A random sample of regulated family day care homes in Texas was examined for compliance with minimum standards. Each was rated on 27 quality aspects that were factor analyzed. Four major factors were identified: environment, caregiver comportment, childcare, and outside relations. Overall, quality and noncompliance were significantly and consistently related. The impact of several specific relationships between quality and noncompliance are discussed from a regulatory point of view, especially with regard to training. PMID- 8404251 TI - In-home family-focused reunification: an experimental study. AB - Intensive, in-home family-based services were employed in reunifying families separated by the placement of a child in out-of-home care. After a 90-day service period, 93% of the 57 families randomly assigned to receive the intensive treatment were reunited-compared to 28% of the 53 families in a control group who received routine out-of-home care services. Follow-up data were collected six and 12 months after the end of treatment. Estimates of outcomes suggest that the experimental treatment had a substantial impact on families and that treatment effects endured for upwards of 12 months following cessation of direct intervention services. PMID- 8404252 TI - Foster care and adoption policy in Romania: suggestions for international intervention. AB - Building an effective child welfare system is a monumental task facing postrevolution Romania. Following revelations on American and Western European television about the large number of "orphans" in Romania, many couples flocked to that country to adopt children. A significant number of adopted Romanian children were brought to the United States. Some of these children are now evidencing problems that are bringing them to the attention of health and social service agencies. This article examines the macroeconomic policies that led to the institutionalization of a large number of children in Romania. Although institutional care is the current norm, a legal basis exists for building family foster care as an alternative. Romania's new adoption law replaces private adoptions with agency-based work. International adoption agencies are involved in developing community-based foster care and permanency planning as part of their work. This article challenges international adoption agencies to use clinical assessments of developmental delays and more rigorous health examinations for children released for adoption. PMID- 8404253 TI - Motor development and the mind: the potential role of motor abilities as a determinant of aspects of perceptual development. AB - Recent advances in the science of human movement have enabled developmental psychologists to discover unique patterns of organization and control in infant motor behavior and development, provoking a resurgence of interest in this topic. In this article, we emphasize the role that motor development may play in determining developmental sequences or "timetables" in other domains. Specifically, we argue that particular motor achievements may be integral to developments in the domains of haptic perception and depth perception. In both cases, there is a high degree of fit between the developmental sequence in which certain perceptual sensitivities unfold and the ages at which the corresponding motor abilities onset. The discussions may provide new contexts in which to consider the developments of haptic perception and depth perception. The general purpose, however, is to highlight the wide-ranging influence of motor development during infancy. PMID- 8404254 TI - Oscillation and complexity in early infant behavior. AB - Cyclic fluctuation is a ubiquitous property of spontaneous motor activity in the human fetus and neonate. Oscillation occurs on the scale of minutes, and irregularity is one of its most characteristic properties. To determine whether cyclic motor activity (CM) persists beyond the neonatal period (1 month after birth), and beyond what has been called the period of the fetus ex utero (2-3 months after birth), 30 infants were studied at monthly intervals from 1 to 4 months after birth during active sleep and awake. Spectral analysis was used to quantify the rate, strength, and irregularity of CM. To examine the characteristic irregularity of awake CM more closely, a nonlinear forecasting technique, developed to study chaotic dynamic in other fields, was used to estimate the predictability of spontaneous movement in a state space reconstructed from the time series. The rate and irregularity of CM during active sleep and awake, and its strength during active sleep, did not change across the first 4 postnatal months. However, there was a pronounced drop in the strength of awake CM from 2 months onward and a concurrent increase in the broad-band power of fluctuations faster than 2 cycles/min. The predictability of awake CM also dropped abruptly between 1 and 2 months, especially for prediction intervals less than 20 sec. The changes at 2 months reflect the introduction of a source of variation with a shorter time constant than fetal-neonatal CM, and which is specific to awake periods. The source of these effects may be moment-to-moment interactions between CM and attention. PMID- 8404255 TI - Development of the stretch reflex in the newborn: reciprocal excitation and reflex irradiation. AB - The stretch reflex is a spinal cord pathway between a muscle's stretch receptors and its own motor units. This reflex is thought to play an important role in normal motor function, because it is unique as a monosynaptic pathway, and because its hyperactivity is a hallmark of many motor disorders. We previously reported a difference in the stretch reflexes in healthy newborn infants and nonambulatory individuals with cerebral palsy (CP): these reflexes are characterized by responses from the stretched muscle and from the reciprocal or antagonist muscle. We proposed the existence of a functional spinal cord pathway that connects both agonist and antagonist muscles at a single joint. We hypothesized that this "reciprocal excitation" is a functional pathway of all newborn infants, which during the normal course of development of motor skills in infants is eliminated. If the CNS is damaged at birth, as in CP, the pathway of reciprocal excitation endures and is reinforced. In the current study of newborns, we recorded stretch reflex responses from all major muscle groups of the lower limb. This "irradiation of reflexes" is a normal phenomenon of the newborn CNS. This pathway becomes suppressed during normal maturation and control of coordinated limb movements. PMID- 8404256 TI - The structuring of neonatal arm movements. AB - The organization and structuring of spontaneous arm movements of 8 neonates were studied quantitatively. The movements were divided up into units, each consisting of 1 acceleration and 1 deceleration phase. This analysis showed that the movements had a distinct temporal structuring. An analysis of curvature was also performed, and it showed that the most distinct changes in movement direction occurred at the transitions between movement units. Finally, the movements of the 2 arms were found to be coupled in all 3 dimensions of space. They had a clear tendency to move together along the body's longitudinal axis, abduct and adduct together, and extend together in the forward direction. PMID- 8404257 TI - The transition to reaching: mapping intention and intrinsic dynamics. AB - The onset of directed reaching demarks the emergence of a qualitatively new skill. In this study we asked how intentional reaching arises from infants' ongoing, intrinsic movement dynamics, and how first reaches become successively adapted to the task. We observed 4 infants weekly in a standard reaching task and identified the week of first arm-extended reach, and the 2 weeks before and after onset. The infants first reached at ages ranging from 12 to 22 weeks, and they used different strategies to get the toy. 2 infants, whose spontaneous movements were large and vigorous, damped down their fast, forceful movements. The 2 quieter infants generated faster and more energetic movements to lift their arms. The infants modulated reaches in task-appropriate ways in the weeks following onset. Reaching emerges when infants can intentionally adjust the force and compliance of the arm, often using muscle coactivation. These results suggest that the infant central nervous system does not contain programs that detail hand trajectory, joint coordination, and muscle activation patterns. Rather, these patterns are the consequences of the natural dynamics of the system and the active exploration of the match between those dynamics and the task. PMID- 8404258 TI - Is visually guided reaching in early infancy a myth? AB - The issue examined was whether infants require sight of their hand when first beginning to reach for, contact, and grasp objects. 7 infants were repeatedly tested between 6 and 25 weeks of age. Each session consisted of 8 trials of objects presented in the light and 8 trials of glowing or sounding objects in complete darkness. Infants first contacted the object in both conditions at comparable ages (mean age for light, 12.3 weeks, and for dark, 11.9 weeks). Infants first grasped the object in the light at 16.0 weeks and in the dark at 14.7 weeks, a nonsignificant difference. Once contact was observed, infants continued to touch and grasp the objects in both light and dark throughout all sessions. Because infants could not see their hand or arm in the dark, their early success in contacting the glowing and sounding objects indicates that proprioceptive cues, not sight of the limb, guided their early reaching. Reaching in the light developed in parallel with reaching in the dark, suggesting that visual guidance of the hand is not necessary to achieve object contact either at the onset of successful reaching or in the succeeding weeks. PMID- 8404259 TI - Visual guidance in infants' reaching toward suddenly displaced targets. AB - This experiment evaluated the role of visual input about the location of a target object and the location of the hand in reaching by infants and adults. 5- and 9 month-old infants were presented with illuminated toys to reach for in a dark room. On no-switch trials, the toy remained illuminated throughout the infant's reach, whereas on switch trials the first-lit toy was replaced during the reach by a second-lit toy at a different position. On approximately half of the trials of each type a luminescent marker was attached to the reaching hand. Adult subjects (tested without the hand marker) fully compensated to the second-lit toy on switch trials, during a second reaching segment. On switch trials, 9-month olds partially adjusted to the second-lit toy when wearing the hand marker and did not adjust without it. On no-switch trials, 9-month-olds reached just as accurately with or without the hand marker. 5-month-olds were generally inaccurate in their reaching and were unaffected by the presence or absence of the hand marker. The findings suggest that during the development of reaching there is an increase in visual guidance during the approach phase of reaches. PMID- 8404260 TI - Infant bouncing: the assembly and tuning of action systems. AB - We outline a theory of infant skill acquisition characterized by an assembly phase, during which a task-specific, low-dimensional action pattern emerges from spontaneous movement in the context of task constraints, and a tuning phase, during which adjustment of the system parameters yields a more energetically efficient and more stable movement. 8 infants were observed longitudinally when bouncing while supported by a harness attached to a spring. We found an initial assembly phase in which kicking was irregular and variable in period, and a tuning phase with more periodic kicking, followed by the sudden appearance of long bouts of sustained bouncing. This "peak" behavior was characterized by oscillation at the resonant frequency of the mass-spring system, an increase in amplitude, and a decrease in period variability. The data are consistent with a forced mass-spring operating at resonance. PMID- 8404261 TI - A longitudinal study of intralimb coordination in the first year of independent walking: a dynamical systems analysis. AB - Using a dynamical systems approach, we examined the development of intralimb coordination over the first year of independent walking. The segmental motion of the thigh and shank and their intralimb coordinative relationship were modeled dynamically as a system of coupled limit cycles. To test the predictions of such a dynamic model in the development of intralimb coordination, 3 infants were filmed from the onset of their first upright independent walking steps, and weekly for the next month and then every month thereafter until they had been walking for 1 year. 3 adults also were included for comparison. Analysis of the kinematic data derived from the film revealed that at the emergence of walking, the limit cycle system displayed instability in several aspects. With walking experience, however, the limit cycle regime became more attractive, such that by 3 months of walking the infants appeared to have found an adult-like stable coordinative relationship between the 2 segments. This pattern of instability followed by stability is consistent with the transitional behaviors of dynamical systems. Several potential control parameters are discussed that might contribute to the stabilization of the walking behavior. The utility of employing a dynamical systems approach for understanding developing behavioral systems is discussed. PMID- 8404262 TI - Crawling versus walking infants' perception of affordances for locomotion over sloping surfaces. AB - 14-month-old toddlers vs. 8.5-month-old crawling infants were encouraged to ascend and descend a sloping walkway (10 degrees, 20 degrees, 30 degrees, and 40 degrees). Infants in both locomotor groups overestimated their ability to ascend slopes. However, on descending trials where falling was more aversive, most toddlers switched from walking to sliding positions for safe descent, but crawlers plunged down head first and many fell at each increment. Toddlers touched and hesitated most before descending 10 degrees and 20 degrees slopes, and they explored alternative means for descent by testing out different sliding positions before leaving the starting platform. In contrast, crawlers touched and hesitated most before descending 30 degrees and 40 degrees slopes, and they never explored alternative sliding positions. In addition, we analyzed measures of locomotor skill and experience in relation to children's ability to perceive affordances. Findings indicate that children must learn to perceive affordances for locomotion over slopes and that learning may begin by fine-tuning of exploratory activity. PMID- 8404263 TI - Commentary: development of perception-action systems and general principles of pattern formation. AB - Our commentary on this special issue devoted to Developmental Biodynamics: Brain, Body, and Behavior Connections is divided into 3 main sections. The first section is an overview of the individual contributions. 5 major themes are identified: (1) inappropriateness of computational treatments of development and the need for more biologically and physically relevant treatments; (2) significance of tailoring muscular to nonmuscular forces in developing movement coordination; (3) importance of spontaneous movements as exploratory and formative mechanisms; (4) influences of action capabilities on the development of perception capabilities, and vice versa; (5) applications of methods and techniques of nonlinear dynamics to developmental processes. In the second section, we provide a synopsis of current ways of thinking about prototypical developmental processes, namely, pattern formation and pattern differentiation, in various classes of physical and biological systems. It is suggested that efforts to understand the progressive formation and differentiation of patterns in terms of very general principles provide a valuable resource of concepts and methods for students of child development. In the third section, hypotheses about the development of perception action systems are generated from juxtaposing the themes and conjectures of this special issue with general principles of pattern formation. The hypotheses suggest the possibility of a pattern formation or dynamics approach to child development as an alternative to the conventional approaches emphasizing maturation (nativist), specific learning experiences (empiricist), cognitive stages (Piagetian), and strategies of encoding and retrieval (information processing). PMID- 8404264 TI - Individual differences in infant visual attention: four-month-olds' discrimination and generalization of global and local stimulus properties. AB - Cognitive performance and development is negatively correlated with fixation duration patterns during infancy, and evidence suggests that long-looking infants may process visual information more slowly than short-looking infants. 3 experiments described here tested the possibility that these differences may be due to differential sensitivity to global and local visual information. Infants were administered discrimination and generalization tasks involving global and local information at varying levels of familiarization time. Results indicated that 4-month-olds process visual information in a global-to-local sequence. Both long- and short-looking infants were sensitive to both types of information, although long lookers required additional familiarization time to match the performance of short lookers. Finally, apparent "generalization" of global information at brief familiarization levels was traced to insensitivity to local stimulus properties. The results do not support the hypothesis that long- and short-looking infants are differentially sensitive to global versus local visual information at 4 months of age. PMID- 8404265 TI - Increasing steps in recall of events: factors facilitating immediate and long term memory in 13.5- and 16.5-month-old children. AB - Children late in the second year of life show patterns of event recall similar to those of older children: (a) well-ordered immediate and delayed recall, and (b) facilitation of recall by familiarity and by enabling relations. We used elicited imitation to test whether the patterns extend to children early in the second year. In Experiment 1, 13.5- and 16.5-month-olds accurately recalled familiar and novel 2-act sequences immediately and after a 1-week delay. For 16.5-month-olds, recall was facilitated by familiarity and by enabling relations; for 13.5-month olds, only enabling relations facilitated recall. In Experiment 2, verbal cues were used to test immediate and 1-week delayed recall of 3-act sequences. For both ages, recall was facilitated by familiarity and by enabling relations. Experiment 3 verified that the verbal information served to cue recall of previously experienced events, not to "suggest" sequences that could be performed. Together the results demonstrate that children as young as 13 months can recall specific events after a delay. They also suggest development in sensitivity to factors that facilitate recall. PMID- 8404266 TI - The development of a linkage between count nouns and object categories: evidence from fifteen- to twenty-one-month-old infants. AB - Recent research suggests that, although young children appreciate many different kinds of conceptual relations among objects, they focus specifically on taxonomic relations in the context of word learning. However, because the evidence for children's appreciation of this linkage between words and object categories has come primarily from children who have made substantial linguistic and conceptual advances, it offers limited information concerning the development of this linkage. In the experiments reported here, we employ a match-to-sample task to focus specifically on the development of an appreciation of the linkage between words (here, count nouns) and object categories in infants in the period just prior to and just subsequent to the naming explosion. The results demonstrate that, for 21-month-old infants, most of whom have recently entered the vocabulary explosion (Experiment 1), and for 16-month-old infants, most of whom have yet to commence the vocabulary explosion (Experiment 2), novel nouns focus attention on taxonomic relations among objects. This is important because it reveals a nascent appreciation of a linkage between words and object categories in infants who are at the very onset of language production. Results are interpreted within a developmental account of infants' emerging appreciation of a specific linkage between count nouns and object categories. PMID- 8404267 TI - Developmental differences in giving directions: spatial frames of reference and mental rotation. AB - The spatial referents "left" and "right" are one of the most common means for specifying direction and location, yet little is known about the processes that underlie the use of these concepts. Two studies tested the hypothesis that children and adults who correctly identify left-right directions from nonoccupied orientations perform imagined rotations to align the self's frame of reference with the other's. Children who make incorrect judgments from nonoccupied orientations were hypothesized to use a stationary egocentric reference frame. 28 6- and 8-year-old children and 9 adults were tested on a task that required making left-right direction judgments from various rotated orientations. The results supported the mental rotation hypothesis: Only correct responders showed a linear increase in response time with increasing angular disparity between the self and other. Mental rotation permits the continued use of an egocentric reference frame for specifying left-right relations from nonoccupied positions. PMID- 8404268 TI - The role of gender-related information and self-endorsement of traits in preadolescents' inferences and judgments. AB - The major purpose of this study was to examine the effects of a target child's gender typicality on different aspects of preadolescents' inferences and judgments. The secondary purpose of the study was to investigate the relation between children's self-endorsement of traits and their inferences and judgments. Fifth and sixth graders were shown a video film, portraying a child playing either a gender-appropriate game with members of the same sex or a gender inappropriate game with members of the other sex. In addition, subjects completed an adapted version of the BSRI and were categorized into sex-typed, androgynous, and undifferentiated subjects. Subjects made a number of different types of judgments and inferences about the target, including inferences about traits, popularity, choice of gift and name, and willingness to engage in activities with the target. All types of inferences and judgments were affected by the variations in the targets' gender-related behaviors, whereas self-endorsement of traits was not related to the inferences and judgments. The results suggest that the gender typicality of the target behavior is salient to preadolescents, regardless of their sex-role orientation. PMID- 8404269 TI - Developmental biodynamics: Brain, body, behavior connections. PMID- 8404270 TI - Developmental biodynamics: brain, body, behavior connections. AB - This special section, Developmental Biodynamics: Brain, Body, Behavior Connections, celebrates a renewed and revitalized interest in the study of motor development. The renewed interest has been sparked by advances in the neurosciences, biomechanics, and behavioral sciences and, importantly, in the attempts to integrate theories and findings across these disciplines. In this introduction to the special section, we consider the contributions that have led to a reinvigorated field of motor development and to its new interdisciplinary look. We highlight the papers in the special section by discussing how they illustrate related advances across research in the neurosciences, biomechanics, and behavioral sciences and how progress across these domains has come to define and characterize the emerging field of developmental biodynamics. PMID- 8404271 TI - Solving Bernstein's problem: a proposal for the development of coordinated movement by selection. AB - In recent years, many established concepts in the theory of human motor development have undergone profound change, and our knowledge has increased greatly. Nevertheless, some outstanding problems remain unsolved. A central problem concerns the redundancy of effective movements, first pointed out by N. A. Bernstein. The human motor system is mechanically complex and can make use of a large number of degrees of freedom. The controlled operation of such a system requires a reduction of mechanical redundancy, effectively by reducing the number of degrees of freedom. More recent work has shown that this problem is hard to solve explicitly by computing solutions to the equations of motion of the system. Equally challenging to traditional computational approaches is the fact the motor systems show remarkable adaptability and flexibility in the presence of changing biomechanical properties of motor organs during development and when faced with different environmental conditions or tasks. Solutions to these problems would have a large impact on a variety of issues in child development. In this article, we stress the importance of the somatic selection of neuronal groups in maps for the progressive transformation of a primary movement repertoire into a set of motor synergies and adaptive action patterns. We present results from computer simulations of a simple motor system that works according to such selectional principles. This approach suggests a provisional solution to Bernstein's problem and provides new parameters to guide experimental approaches to the development of sensorimotor coordination. PMID- 8404272 TI - Biomechanics and developmental neuromotor control. AB - By applying the principles and methods of mechanics to the musculoskeletal system, new insights can be discovered about control of human limb dynamics both in adults and infants. Here, we first provide a basic primer about biomechanics- its historical context, terminology, and analytical techniques. Next we review research with animals and human adults that illustrates how limb dynamics provides a window for examining the physical mechanisms underlying neuromotor control. Finally, we elaborate on how our research group has adapted dynamics techniques to investigate how infants gain control of their limbs and learn to reach in the first year of life. PMID- 8404273 TI - Evaluation of LDL subfractions in vascular complications of familial hypercholesterolemia. AB - The authors attempted to investigate the possibility of estimating modified LDL (acetylated LDL, oxidative LDL and glucosylated LDL) with peroxidized lipid in the native LDL from patients with familial hypercholesterolemia (FH). A group of 7 patients with FH were studied, with age ranging from 18 to 60 years. 10 ml of fasting blood were collected into siliconized syringes with 0.5 ml of 0.05 M ethylene diamine tetraacetic acid 2Na. LDL, LDL1 and LDL2 were separated by sequential ultracentrifugation. Total cholesterol (TC), triglycerides (TG) and phospholipids (PL) were estimated by enzymatic methods. Analyses of lipids in each LDL were done by thin layer chromatography (TCL). The modified LDL in native LDL of the FH patients was estimated by the authors' own method. The cholesterol concentration of LDL was 0.49 +/- 0.28 mmol/L in FH patients and 0.07 +/- 0.04 mmol/L in the healthy persons (P < 0.001). The LDL1 cholesterol concentrations in the patients with FH and the normal controls were similar while that of the LDL2 was of significant difference (P < 0.05). On the TLC plate, the detection of the spot X1 was considered an indicator of the existence of modified LDL. The percentages of spot X1 in LDL of FH patients were higher in all 3 fractions (LDL, LDL1, and LDL2), as compared to the corresponding percentages of spot X1 in the healthy controls. (P < 0.001). Using the authors' own method, the molecular size were found in the order of LDL > LDL1 > LDL2. PMID- 8404274 TI - Serum tumor necrosis factor (TNF) in the pathogenesis of clinical hepatic failure of HCV and/or HBV infection. AB - Serum TNF and IL-6 levels were measured in 48 patients with liver disease positive for anti-HCV only or concurrent HBV infection. High serum TNF levels were observed in patients with liver disease positive for anti-HCV and/or HBV infection (P < 0.001). Serum TNF levels varied with the severity of liver disease. Serum TNF levels of anti-HCV positive patients with hepatic failure were higher than those with CAH (P < 0.01). Serum TNF levels of patients infected with HCV or concurrent HBV were also significantly higher than those with HBV infection alone (P < 0.001). However, no difference in serum IL-6 levels was observed in either group of patients. Serum TNF in the deceased patients with hepatic failure induced by HBV and HCV infection was higher than in those who survived (P < 0.05), and it also seemed significantly different in patients with and without multiple organ failure (P < 0.05). In vitro, HSS showed marked inhibitory activity on TNF production from PBM induced by endotoxin, but had no significant effect on the TNF cytotoxicity of L929 cells. It seems that high serum TNF level is an important mediator in the pathogenesis of liver necrosis and failure of microcirculation in HCV and/or HBV infection. These observations favor the attempt to treat hepatic failure with HSS or anti-TNF. Encouraging results were achieved using HSS in the treatment of subacute liver necrosis in our institute.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404275 TI - Surgical correction of hypertelorism. Report of 40 cases. AB - Forty patients with hypertelorism seen in the past 16 years were reviewed retrospectively. Combined intra- and extracranial surgical approach was used for 37 severe and moderate cases and subcranial approach (U-osteotomy) for 3 moderate cases. Gratifying results were obtained in patients with different types of hypertelorism by a multidisciplinary team. Complications were also reviewed. Of the 37 cases of intra- and extracranial corrections, 1 died, 4 had cerebrospinal fluid leakage and 4 had keratitis. No seizure, cerebral edema, meningitis, blindness, and ptosis occurred in this series. The average age was 13 years and two months. Operating time averaged 6 hours and 50 minutes. Hypertelorism was mostly attributable to craniofacial cleft, craniosynostosis, frontoethmoidal meningoencephalocele, frontonasal fibrous dysplasia, and trauma. Satisfactory esthetic appearance was achieved in most of the cases. PMID- 8404276 TI - Target therapy of ovarian carcinoma by monoclonal antibodies bearing chemical drugs entrapped in liposomes. AB - Monoclonal antibody COC166-9 against ovarian carcinoma was conjugated with adriamycin or cisplatinum entraped in liposomes as immunochemical liposomes MLA and MLP respectively. MLA was shown to have the highest effect than adriamycin or other groups on SKOV3 (ovarian cancer cell line) growth inhibition. MLA was also used in target therapy on nude mice bearing human ovarian carcinoma by xenograft of SKOV3 cells. The observations of tumor volumes revealed that this target therapy other than the controls presents a significantly better result which gives a hopeful clue to ovarian carcinoma treatment. PMID- 8404277 TI - Research on hemodynamics of cerebral arteriovenous malformation by Doppler ultrasound. AB - Combined extracranial and transcranial Doppler (TCD) instruments were used to study the hemodynamics of 20 patients with cerebral arteriovenous malformation (AVM) proved angiographically. It was discovered that the flow velocities in AVM feeding arteries and their proximal arteries were increased; the increase of flow velocities in feeding arteries close to AVM was more remarkable than that in feeding-arteries proximal to extracranial internal carotid artery far from AVM; pulsating indexes of AVM-feeding arteries were decreased; increase of flow velocities in AVM-feeding arteries was related to the decrease of pulsating index; the flow velocities of nontapering feeding arteries were faster than those of tapering arteries; the greater the size of AVM, the faster the velocity in feeding artery; and the flow velocities and pulsating indexes of AVM-feeding arteries were gradually recovered to normal 3-5 weeks after resection of AVM. It is believed that increase of flow velocity in AVM-feeding artery is associated with distention and decreased resistance of flow in AVM-feeding artery. TCD combined with CT scans are helpful to the diagnosis of AVM. Combination of TCD and cerebral angiography (CAG) to evaluate comprehensively the preoperative hemodynamics of AVM and to monitor postoperative changes is helpful to detecting cerebral steal due to steal leakage in circle of Willis and preventing the hazardous postoperative complications caused by pressure breakthrough of normal perfusion. PMID- 8404278 TI - Air bronchogram in differential diagnosis of small peripheral lung cancers on CT image. AB - The computed tomographic (CT) images of 52 patients of small peripheral lung nodule < 3 cm proved pathologically were reviewed. Air bronchogram (AB) was found in 14 nodules on thin-section CT images, in which well or moderately differentiated small adenocarcinomas were only seen with lipidic growth. Squamous cell, large-cell carcinomas and small benign lung nodules had no AB sign both on CT image and in pathologic specimens. PMID- 8404279 TI - Distribution of materno-fetus materno-suckling transfer of tritium after single injection of tritiated water in mice. AB - The metabolism and transfer of tritium from pregnant mice to fetus after injection of tritiated water intraperitoneally was investigated in this paper. The study was composed of three experiments. 1. Pregnant mice were injected with tritium water on the first day of pregnancy, in order to obtain a transfer coefficient of tritium from pregnant mice to fetus through placenta. 2. Pregnant mice were injected with tritiated water on the first day of parturition to study the transfer of tritium via mother milk from nursing mice to the baby-animals. 3. Pregnant mice were injected with tritiated water in different periods of gestation. The results showed that tritiated water was uniformly distributed in all of the tissues measured, including placenta, fetal membrane and amniotic fluid in experiment 1. No effect of placentas on tritium transfer from pregnant mice to fetus was found. Concentration of tritium in the baby's tissues was evidently higher than that in the pregnant mice in experiments 2 and 3, and the transfer coefficient in experiments 2 and 3 was generally higher than that in experiment 1. PMID- 8404280 TI - CD38- and Leu6-positive cells in dermal lesions of mycosis fungoides. AB - The phenotypes of mononuclear cells in 16 patients with mycosis fungoides (MF) (8 were in tumor stage and 8 in pretumor stage) were tested by ABC immunochemical technique. The results showed that the percentage of CD38 and Leu6(+) cells in tumor stage was higher than that in pretumor stage. CD38 was identified to be a marker of the immature lymphocytes. The change of CD38 might be helpful in staging MF and predicting the prognosis of the disease. In addition, Leu6(+) cells, oval or round in shape and without dendritic processes were found in tumor stage, suggesting that they might be immature lymphoid cells. This feature needs further investigation. PMID- 8404281 TI - Effective response evoked by microstimulation of thalamus nuclei in patients with tremor. AB - 459 microstimulations were done in 10 patients with tremor. The motor evoked response with reduction tremor drive was observed at 38 sites, of which 30 (79%) were noted in ventrointermedialis (vim) nuclei, and 5 (13.2%) in ventrocaudalis (vc) nuclei, 2 in ventroralis posterior (vop) nuclei, 1 in dorsal thalamus (dth). No increased tremor drive response was noted in all the sites. Paresthesia was the most common response (47.1%). Except pain at 1 site in vc nuclei, warm/cold and vertigo were noted in vop, vim, vc nuclei. No responses accounted for 43.8%. PMID- 8404282 TI - Progress in molecular medicine. From experimental genetics to the treatment of hypertensive patients. PMID- 8404283 TI - An improved technique for bloodless hepatic resection on in situ cold perfused liver. AB - An improved technique for bloodless hepatic resection using in situ isolation and asanguinous hypothermic perfusion was described to deal with huge liver tumors involved in the liver hilum, the main hepatic veins and retrohepatic inferior vena cava. The original Fortner's technique was modified, including the choice of incision; semi-isolated perfusion of the liver portion preserved through the single portal vein; suprahepatic outlet of the perfusate and the shortening of the period of hepatic ischemia by reperfusion of hepatic artery prior to the repair or reconstruction of the portal vein. The initial successful experience of the technique applied to 2 pediatric cases with giant liver tumors was reported, and the indications, intraoperative and early postoperative courses were discussed. PMID- 8404284 TI - Evaluation of hypertension and other risk factors in ischemic heart disease. AB - The author presented data obtained in Bangladesh, to elucidate the role of hypertension as a risk factor along with others such as smoking, cholesterol and diabetes mellitus as cofactors in ischaemic heart disease (IHD). There was a series of 100 cases with IHD admitted within 12 hours after the onset of chest pain observed in this study. They all were diagnosed as IHD for the first time. Of them, 94 were male and 6 female, with an age range of 25-77 years (mean 50.16 +/- 14 years). On grouping of IHD, 21 had angina pectoris and 79 acute myocardial infarction. 31% cases of IHD had hypertension. The blood pressure ranged between 168.54 +/- 24.85 and 106.29 +/- 16-80 mmHg. 74 out of the 100 cases with IHD were smokers. The mean value of serum cholesterol in this series was 6.48 +/- 1.66 mmol/L and that among 50 normal controls was 4.76 +/- 1.28 mmol/L (P < 0.01). The serum triglyceride determinations between the 94 cases of IHD and 50 normal controls showed values with statistically significant difference. The author concluded in this study that, hypertension, smoking and hyperlipidemia are the most important risk factor of IHD in Bangladesh. PMID- 8404285 TI - [Ambulatory surgery in the hospital. Perspectives and problems from the medical organizational viewpoint]. PMID- 8404286 TI - [Trust in medicine, especially in surgery]. PMID- 8404287 TI - [Continence preserving operation after proctocolectomy. Indications, technique and results]. PMID- 8404288 TI - [Direct ileum-pouch-anal anastomosis]. PMID- 8404289 TI - [Reconstruction of continence after colectomy--a determination of current status. Indications, technique and results]. PMID- 8404290 TI - [Thoracoscopic surgery of the lung and pleura]. AB - 134 operations were performed using video-thoracoscopic techniques for patients, with diffuse lung disease (48), pneumothorax (46), peripheral coin lesions (29) und others (11). In 92% of the patients video-thoracoscopic operation was completed. Open reoperation was indicated only in patients with persistent pneumothorax (3). After hospital discharge open reoperation was performed once for recurrent pneumothorax. From our experience we conclude, that thoracoscopic surgery may be routinely indicated in future for lung biopsies, pneumothorax, peripheral coin lesions and pleural products. PMID- 8404291 TI - [Surgery for inguinal hernia in Germany 1992: a survey of 1,656 German clinics]. AB - A questionnaire of 1656 German surgical departments shows the preferentially applied procedures: in hernia of childhood Rehbein/Grob (24.1%) and Gross/Ferguson (24.4%). In primary hernia of adults Shouldice repair (47.7%), in recurrent hernia Shouldice-repair as well. McVay, mesh or plug-techniques are applied only for problematic cases of recurrent hernia. Laparoscopic hernia repair is accepted in less than 10% of clinics. PMID- 8404292 TI - [Broad use of laparoscopic cholecystectomy in the teaching clinic. Experiences and results of 300 operations]. AB - Laparoscopic cholecystectomy (LC) was integrated early in surgical training according to the concept "observe, assist and then operate under the assistance of an experienced surgeon". 300 LC's were performed by 27 different surgeons (8 board certified surgeons, 19 residents in the year 2-6 of training). 60.3% of all LC's and 73% of LC's performed for acute cholecystitis were done by residents. Our results including an overall complication rate of 5.3% shows, that LC can be performed by residents without additional complications or unacceptable length of operations, provided the threshold for conversion is low and the surgeon is assisted by an experienced laparoscopist. We therefore suggest that the early integration of LC in surgical training is justified. PMID- 8404293 TI - [Perioperative use of ampicillin/sulbactam, cefoxitin and piperacillin/ metronidazole in elective colon and rectal surgery. A prospective randomized quality assurance study of 422 patients]. AB - As has been proved before, antibiotic prophylaxis is highly effective in lowering wound infection rates in colorectal surgery. In order to establish quality control, we checked the effectiveness of three different prophylactic antibiotic regimes in 422 patients in a prospective and randomized trial. Between the three groups were no significant differences as regards age, type of operation and risk factors like adipositas and diabetes. The wound infection rate according to CDC criteria was from 7.0 to 9.5%. We did not find a significant difference between the three antibiotic regimes. It is therefore our conclusion, that in our setting each of the three different types of antibiotics is of equal value. This means, on the other hand, that the cheapest one is enough. PMID- 8404294 TI - [Syndrome of solitary rectal ulcer--a rare cause of recurrent rectal hemorrhage]. AB - While the clinical symptoms and signs of SRUS are quite obvious, etiology and treatment of the disease remain to be obscure. Although the lesion is usually not solitary this name has been used for the disease. According to other authors, we also find this term inappropriate. During the last 5 years 6 cases of SRUS have been diagnosed, treated and prospectively assessed in the surgical clinic of Cerrahpasa Medical Faculty (Istanbul). Three patients were treated successfully by conservative means, transanal excision was performed and defects were sutured primarily in the other three. In one patient recurrence was seen 8 days after the transanal excision; transabdominal rectopexy was performed 6 months later. All patients have been followed up with no complaints. PMID- 8404295 TI - [Post-primary functional soft tissue coverage after extensive muscle-/soft tissue loss of the upper extremity]. AB - Two cases of severe contusion/avulsion injuries of the upper extremity with involvement of the neuro-vascular structures and extensive muscle necrosis after post-ischemic syndrome (PIS) are demonstrated. The concept of early post-primary soft tissue coverage and the opportunity of a simultaneous "functional" soft tissue replacement with pedicled neuro-vascular myocutaneous island flaps is elucidated. PMID- 8404296 TI - [Cushing syndrome in ACTH-producing neuroendocrine tumor of the hepatic duct]. AB - We present a patient with an ACTH producing neuroendocrine tumor of the hepatic bile duct which was detected by chance during abdominal surgery for Cushing's syndrome. Diagnostic strategy and surgical therapy in patients with neuroendocrine tumors are discussed. The diagnostic problems caused by combination with an endocrine function disorder like Cushing's syndrome are pointed out. PMID- 8404297 TI - [Incarcerated scar hernia in an adjacent channel after laparoscopic cholecystectomy--a rare complication?]. AB - The rare case of a postoperatively incarcerated cicatricial hernia in an auxiliary instrument channel after laparoscopic cholecystectomy is reported. To avoid such a complication one should: 1. Oblique insertion through the abdominal wall of all operation trocars. 2. Placing the lateral auxiliary instrument channel through the straight abdominal muscle under consideration of the epigastric vessels instead of going through the white line. PMID- 8404298 TI - [Progress on pathological study of ovarian epithelial tumors]. PMID- 8404299 TI - [Summary of the Third National Conference on Gynecological Pathology]. PMID- 8404300 TI - [Histopathologic analysis of mullerian adenocarcinomas]. AB - A histopathologic review of 959 cases of the endocervical, endometrial and tubal adenocarcinomas is presented. Most of the histopathologic types of the adenocarcinoma in each site were in consistent with that of it's native epithelium, which were 43.7%, 74.5% and 85.5%, respectively. However, other types besides native type may be found in any site adenocarcinoma. The pathologic features of the surroundings in some early lesions demonstrated a possible different morphogenesis. The synchronous multiple site primary adenocarcinomas in the mullerian system were common, comprising 7.8%, in which were composed of 91% in 2 sites, 7.5% in 3 sites, and 1.5% in 4 sites. Most of them were endometrial and ovarian adenocarcinomas, in 37.3%. These data suggested the complex histopathologic types of the mullerian adenocarcinomas including the endocervical, endometrial and tubal adenocarcinomas should be considered before diagnoses, especially in biopsy specimens. The synchronous multiple site primary adenocarcinomas were not rare for the same origin of histogenesis. Thus, a correct diagnosis will be conductive to stage and management. PMID- 8404301 TI - [Immunohistochemical study on human papillomavirus infection of the vulva]. AB - Two hundred and sixteen cases of human papillomavirus infection of the vulva from 1984 to 1991 in the Affiliated Hospital of Qingdao Medical College were reviewed, and were immunohistochemically studied by ABC method to detect HPV-Ag. The results showed that the demonstration of diagnostic koilocytes is very important in diagnosis of HPV infection in routine tissue slides examination. But in cases of atypical morphological changes; when diagnostic koilocytes were not formed in early stage, the demonstration of brown color granules in the nuclei of prickle cells is very diagnostic for positive HPV infection. In occasional cases, the diagnostic koilocytes do not demonstrate brown color granules in their nuclei. The explanation is that HPV-Ag was exhausted during metabolism. Besides, the cell membrane of basal cells are stained with brown color granules, while the morphological changes of upper layers of squamous epithelium have not appeared yet, therefore, there were no HPV-Ag positive reactive cells. It is probably showed that the HPV-Ag is primarily formed and appeared in the cell membrane of basal cells. PMID- 8404302 TI - [Diagnostic value of electron microscopy on epithelial tumors of the ovary]. AB - The study on transmission electron microscopy (TEM) and scanning electron microscopy (SEM) in 52 cases of ovarian epithelial tumors (OET) was presented. The characteristic features of ultrastructure and cell surface structure of different types of OET, also the diagnostic features of cancer cells were determined and discussed. It has a great value in differentiation of histologic types of OET and in early diagnosis of cancer. In 3 cases of borderline malignant tumor, some malignant cells, which were impossible to distinguish by light microscopy, were discovered by TEM and SEM. The diagnostic value of electron microscopy in early malignancy of tumors is more important. PMID- 8404303 TI - [Colposcopical and pathological study of vulvar pseudocondyloma]. AB - One hundred and forty seven cases of pseudocondyloma, which were diagnosed by colposcopy and histopathology, and 129 cases of condyloma as control group were studied. Pseudocondyloma showed colposcopically as crowded together distribution of single papilla or vesiculated papules without branching structure and capillary loops. The color of the lesions is unchanged after acetic acid stain. The lesions can be divided into vesicular proliferative type and ciliary proliferative type. Differing from condyloma, pseudocondyloma lesions showed histopathologically only slight proliferation of squamous epithelium, lack of diagnostic koilocytes and negative result in HPV immunohistochemical study and hybridization of HPV-DNA. It is a reliable method to make a diagnosis of pseudocondyloma by colposcopy and histopathology. PMID- 8404304 TI - [Ultrasonic color Doppler flow imaging detect coiling of the umbilical cord around fetal neck]. AB - Twenty eight third trimester pregnant women that B mode ultrasound showed coiling of umbilical cord around fetal neck (UCAN) were monitored by Color Doppler Flow Imaging (CDFI) before delivery. UCAN were discovered in 12 women by CDFI, and all were confirmed at delivery. The accurate diagnosis rate was 100%. The accuracy rate of "no UCAN" diagnosed by CDFI was 93.75%. The accurate diagnosis rate of CDFI was significantly higher than that of the simple B mode ultrasound examination; and the misdiagnosis rate of "no UCAN" was 6. 25%, significantly lower than B mode (53.57%) (both P < 0.01). CDFI might influence the mode of delivery and the fetal prognosis. This study showed that CDFI can precisely detect whether there is UCAN or not before delivery. Meanwhile, the diagnosis standards of UCAN was established. PMID- 8404305 TI - [Cardiotocography admission monitoring and intermittent intrapartum monitoring for fetal distress in labour]. AB - Cardiotocography (CTG) monitoring on admission was performed in 71 patients for 30 to 60 minutes in a semilateral position. Intermittent intrapartum CTG monitoring were performed in 40 of these patients. Admission monitoring might provide information of fetal basic status and predict asphyxia that could develop during labour. There was less intrapartum asphyxia, lower Apgar score, umbilical artery pH < or = 7.10 and meconium stained amniotic fluid in high-score group than that in medium-score group and low-score group (P < 0.05). Admission monitoring had high predictive value of umbilical cord complication in labour and enable to detect them in the early first stage of labour. Fischer's composite score of FHR of admission monitoring was closely related to that of intermittent FHR monitoring in the late first stage of labour. In this study, it is suggested that admission monitoring is a useful, simple, convenient and noninvasive method for screening fetal basic condition. When continuous FHR monitoring is not available, the intermittent intrapartum FHR monitoring in the late first stage of labour is of importance. PMID- 8404306 TI - [Long-term follow-up of the ovarian malignant teratoma]. AB - Sixty-six patients with ovarian malignant teratoma have been treated in our hospital from Jan. 1954 to Dec. 1990. Two histological types were covered: (1) some malignant tissue came from mature teratoma; (2) embryonic tissue immature ovarian teratoma. During the process of long-term follow up, 38 cases survived, 28 died. With life table calculation, the 5, 10, 15, 20, 25 and 30 year survival rates of the malignant teratoma were 61.5 +/- 6.1; 57.8 +/- 6.2; 57.8 +/- 6.2; 52.8 +/- 7.4; 52.8 +/- 7.4; and 52.8 +/- 7.4%, respectively. The prognosis was related to the age of patient, the size of the tumor, the histological grade and especially, to the clinical staging. The treatment method of this teratoma was discussed as well. PMID- 8404307 TI - [Pharmacokinetics study of intraperitoneal cisplatin in ovarian carcinoma]. AB - Ten patients with surgically and pathologically confirmed ovarian cancer were treated by intraperitoneal (IP) chemotherapy of 20-25 mg/m2 of cisplatin. Total platinum concentrations in plasma, ascitic fluid and urine were determined by flameless atomic absorption spectrophotometry. The peak of total platinum concentration in ascitic fluid was 18.75 times greater than that in plasma. The half-life of the elimination for total ascites platinum and total plasma platinum were 39.9 hours and 70.6 hours respectively. Peritoneal clearance of cisplatin was 6.1 ml/min whereas body clearance of cisplatin was 15.3 ml/min. 24 hours following administration, 13.53% of cisplatinum had been excreted through the urine. The results indicate that there is pharmacokinetic advantage to be gained by using IP chemotherapy of cisplatin. Because the half-life of cisplatin in plasma is longer, therefore, the interval of chemotherapy should be three weeks or more. PMID- 8404308 TI - [Double primary carcinoma of the uterine corpus and ovary]. AB - This article presents 19 patients with double primary carcinoma of the uterine corpus and ovary diagnosed and treated at Queen Mary Hospital, University of Hong Kong in last ten years. 14 patients had typical endometrial adenocarcinoma and endometrioid carcinoma of the ovary (group A), 5 had non-endometrioid carcinoma in the uterine corpus and/or ovary (group B). There is no significant difference in survival rate in group A and B (78.6% and 80%, respectively, mean follow-up 42 months), but the total survival rate in our series is 78%, which is equal to stage I ovarian cancer. The tumor grade and positive abdominal washing were proven to be statistically significant indicators of the poor prognosis (P < 0.05). The result reveals that the prognosis of double primary carcinoma of the uterine corpus and ovary is better and it is necessary to distinguish double primary carcinoma of the uterine corpus and ovary from stage II ovarian cancer and stage III endometrial carcinoma. PMID- 8404309 TI - [Blood rheology in perinatology]. PMID- 8404310 TI - [Thrombotic thrombocytopenic purpura in pregnancy]. PMID- 8404311 TI - [Development of the high effective thrombolytic agents with molecular biological technique]. PMID- 8404312 TI - [Quantitative assessment of the effect of nitroglycerin on left ventricular function with two-dimensional echocardiography in patients with myocardial infarction]. AB - The effect of sublingual and intravenous nitroglycerin (SLNTG and IVNTG) on left ventricular global and segmental systolic function was quantitatively assessed with two-dimensional echocardiography (2DE) in 28 cases of myocardial infarction (MI), including a group without heart failure of 13 and another group with heart failure of 15. The results showed that after SLNTG and IVNTG the left ventricular end-diastolic and end-systolic volumes were significantly decreased in both groups (P < 0.05-0.001), the left ventricular global systolic function was improved particularly in MI patients with heart failure during IVNTG. The wall motion in the normal and/or hypokinetic segments was enhanced after both SLNTG and IVNTG. It was shown that nitroglycerin could strengthen the left ventricular global systolic function in patients with MI. PMID- 8404313 TI - [Classification and treatment of torsades de pointes (TdP): a report of 11 cases]. AB - A clinical analysis of 11 cases with TdP was reported. These patients were divided into two groups: 9 cases with long QT syndrome and 2 with normal QT interval. All of the patients suffered from coronary heart disease with various concomitant ailments, such as pulmonary heart disease (1 case), atrioventricular block (5 cases with complete or type II second degree AV block), hypokalemia (7 cases). In one case the TdP was caused by aminodarone administration. The principle of treatment included removing the inducing causes, treating the primary diseases and interrupting TdP. All patients who had hypokalemia were treated with potassium chloride. 7 of 9 cases showed effective response to treatment with magnesium sulfate. The patients with long QT syndrome failed to respond with MgSO4. Severe bradycardia should be treated with isoprenaline. Only one patient had pacing therapy and another received electric conversion. According to our experience, for patients of TdP with long QT syndrome class Ia and III antiarrhythmic agents were contraindicated, but those with normal QT interval can take these drugs. PMID- 8404314 TI - [A comparison between diltiazem SR and diltiazem HCL in the treatment of angina pectoris]. AB - This study included 168 patients with angina pectoris, who were divided into two groups, the diltiazem SR (No. 88) group and diltiazem HCL (No. 80) group. The two groups were comparable in age and duration of coronary heart disease. The results were as follows: (1) The total clinical effect of diltiazem SR (94.3%) was slightly greater than that of diltiazem HCL (82.5%), the effect in ECG improvement was 53.3% and 53.4% respectively. (2) The rate of adverse effect was 12.5% in diltiazem SR group and 14.4% in diltiazem HCL group. It is concluded that the two different types of diltiazem have similar effect in the treatment of angina pectoris. However, it is more convenient to take diltiazem SR than diltiazem HCL as the former needs only twice a day. PMID- 8404315 TI - [The nutritional status and immune function of patients with chronic obstructive pulmonary disease]. AB - The nutritional status and immune function of 14 patients with exacerbation of chronic obstructive pulmonary disease (COPD) were assessed and were studied again one week after the patients had recovered. The results showed that the nutritional status of the patients was poorer on admission, the level of some kinds of serum free amino acid was significantly lower than the normal value, the value of branch-chain amino acid (BCAA) was significantly lower than that in healthy people (P < 0.01), but the level of aromatic amino acid (AAA) was higher than the normal value. Level of serum free fatty acid was significantly lower during admission and in recovery period (P < 0.01). It is suggested that shortage of essential fatty acid in the patients is obvious. The results also showed that the patients identified as malnourished had a lower cellular immune function, but the humoral immune function was not significant between the patients group and control group. Immune function was improved with the recovery nutritional status of the patients. The results suggest that malnutrition and hypoimmunity may play a very important role in the pathogenesis of COPD. PMID- 8404316 TI - [Effect of ligustrazine controlled release capsule on hemorrheology in patients with cor pulmonale]. AB - Before and after oral administration of Ligustrazine controlled release capsule, changes of hemorrheology and TXA2/PGI2 were evaluated in 16 patients with advanced cor pulmonale. It was shown that the effects of decreasing whole blood and plasma viscosity and lowering hematocrit and fibrinogen level were noted after one course of treatment with this medication. It is suggests that anticoagulant, effect of Ligustrazine is probably related to its role of improving inbalance between TXA2 and PGI2 in patients with cor pulmonale. PMID- 8404317 TI - [The renal involvement of primary vasculitis: clinical and pathological analysis of 5 cases]. AB - Primary vasculitis, include microscopic polyarteritis (MPA) and Wegener's granulomatosis (WG), is one of the causes of Glomerulonephritis and renal failure. Recently antineutrophil cytoplasmic autoantibodies (ANCA) is considered to be a very important tool for the diagnosis of MPA and WG. The management of ANCA was performed in 42 patients with variant primary glomerulonephritis with crescents formation or sclerotic glomerulonephritis, 5 of them show positive, appeared as a pattern of P-ANCA. Most of the patients were old male, with fever, malaise, weight loss, anemia, muscle and joint pain, some of them have chronic cough, even hemoptysis, opacities in middle-lower fields of both lungs, which didn't response to antibiotics therapy. This is the first report about renal involvement of primary vasculitis from China. Our result suggest that primary vasculitis is not a rare course of renal failure in China. PMID- 8404318 TI - [The diagnostic accuracy of gastric urease activity for Helicobacter pylori]. AB - The diagnostic accuracy of gastric urease activity for Helicobacter pylori (HP) infection were studied in 98 patients with peptic ulcer, gastritis and chronic renal failure. Gastric aspirates were analyzed for urea nitrogen and ammonia. Urease activity was calculated as the ratio of gastric ammonia to sum of gastric urea nitrogen and ammonia. Gold standard assessments to HP infection were rapid urease test, histology and/or culture. The results showed that the values of urease activity in patients with HP infection higher than that in patients without HP infection. This were further assessed by eradication of HP. Sixteen patients with HP infection were treated with DeNol 330 mg/day for 4 weeks. HP was eradicated in 8/16 patients. In those cases urease activity reduced (P < 0.05). If taken 0.31 as the cut-off index of urease activity to diagnosis HP infection, the sensitivity, specificity and accuracy were 95%, 84%, 93% respectively and were not affected by blood urea nitrogen. PMID- 8404319 TI - [Study of cytopathogenic effect of epidemic hemorrhagic fever virus on normal human bone marrow cells]. AB - The cytopathogenic effect of epidemic hemorrhagic fever virus (EHFV) on cultured human bone marrow cells (HBC) was studied. Specific viral antigen in HBC was detected at various times after EHFV infection with trypan-blue stain and electron microscopy. It was shown that the infection rate of HBC was over 80% at 3 weeks after infection. The death rate of HBC in the infection group was significantly higher than that in the control group on 1-4 weeks after infection. Injury of cell membrane organs after infection with EHFV were significantly earlier and more severe as it was shown by electron microscopy. It was demonstrated that EHFV could cause cytopathogenic effect on cells of nomal human bone marrow. PMID- 8404320 TI - [Treatment of congestive heart failure complicating chronic Keshen disease with magnesium sulfate and captopril]. AB - 150 hospital patients of chronic Keshen disease with congestive heart failure were divided into two groups: the treatment group (TG) and control group (CG). Patients in TG (80 patients) were treated with magnesium sulfate and captopril, while those in CG (70 patients) with digoxin. The results showed a marked difference (P < 0.01) in the total effective ratio between TG (92.5 percent) and CG (65.7 percent). Both groups had an increase in cardiac output and cardiac index, but it was more apparent in TG. With sum vessel resistance (SVR), TG had a remarkable decrease (P < 0.01), patients in while those in CG did not (P > 0.05). The results also showed that the therapeutic effect in TG was satisfactory. The important function of magnesium, the basis of its treatment and the simultaneous administration of magnesium and captopril to reduce the cardiac preload and afterload and to improve the heart function were discussed. PMID- 8404322 TI - [The prediction of thrombotic diseases]. PMID- 8404321 TI - [Thrombolytic therapy of acute myocardial infarction]. PMID- 8404323 TI - [Clinical studies of recombinant human erythropoietin in patients on long-term dialysis]. AB - Recombinant human erythropoietin (r-HuEPO) was administered in 68 dialyzed patients (32 on acetate hemodialysis, 24 bicarbonate hemodialysis and 12 on hemo filtration). The mean initial Hb 52.7 +/- 8.0g/L, Ht 19.4 +/- 2.2%, serum ferritin > 100ng/L. Each patients received r-HuEPO intravenously, at the dose of 300U/Kg/w for 6.2 +/- 4.3 months. Target range: Hb 100-120g/L, Ht 30-35%. After r HuEPO treatment, blood transfusion was not needed for any of the patients, anemia was ameliorated with increase of Hb and Ht levels. It was found that the minimum effective dose of the r-HrEPO was 150-300U/Kg/w. We conclude that r-HuEPO is effective as treatment for the anemia of dialyzed patients. However, hypertension, clotted dialyzers and dialysis access thromboses were been developed in some patients after correction of anemia. There is now a general consensus that these side effects may be minimized if r-HuEPO is initially given in small doses with increments to avoid a too rapid correction of the anemia. PMID- 8404324 TI - [The changes of right ventricular function in the course of COPD induced cor pulmonale]. AB - A study on pulmonary hemodynamics, radionuclide right ventricular ejection fraction and blood gas analysis in 62 cases of COPD induced cor pulmonale at different stages showed the following results: 1. In early stage of cor pulmonale, stroke volume could not increase after exercise, it proved that cardiac reserve has been reduced; 2. With the progression of the disease, cardiac stroke volume reduced but cardiac output increased gradually, it could be considered as the evolutionary characteristic of cardiac function in chronic cor pulmonale; 3. The right ventricular stroke work was normal and could increase with the rise of after-load, reflecting the relatively effective functional compensation; 4. In acute exacerbation of cor pulmonale, the cardiac failure should be attributed to hyperdynamic type with hypervolemia; 5. Correlation analyses suggested that cardiac output decreased along with the increase of right ventricular afterload only in acute exacerbation of late cor pulmonale; PaO2 and PaCO2 have only slight influence on right ventricular function. PMID- 8404325 TI - [The purification of SS-B antigen and detection of anti-SS-B antibodies]. AB - SS-B antigen was purified from fresh rabbit thymus by ammonium sulfate precipitation and column chromatography with Sephadex G100 and phosphocellulose. The M. W. of SS-B is ranged at 41,000 to 48,000. It does not contain the other extractable antigens, like Sm, RNP, PM-ScL, Scl-70, Jo-1, and PCNA. The purified SS-B antigen only reacts with the CDC standard serum of anti-SS-B antigen only reacts with the CDC standard serum of anti-SS-B antibody by ELISA. The positive rate of the antibodies being 55.1%, 48.3%, 32.8%, 30.8% and 26.3% in SS, SLE, RA, PSS and MCTD respectively. The titers of anti-SS-B antibodies were higher in SS and SLE patients than other connective tissue disease patients. It was found that all of the anti-SS-B antibodies detected were mainly of IgG isotype. Preliminary analysis of clinical date shows that there is no relationship between anti-SS-B antibody and systemic involvement in SS. PMID- 8404326 TI - [Human parvovirus B19 and its associated diseases]. PMID- 8404327 TI - [Clinical application of bone marrow transplantation]. PMID- 8404328 TI - [Prevention of alloimmunization is first priority in platelet transfusion]. PMID- 8404329 TI - [Phase II clinical trial of domestic amsacrine in patients with acute leukemias. Cooperative Group for Treating Acute Leukemias with Domestic m-AMSA]. AB - 118 patients with acute leukemias, including initial, relapsed and refractory cases, were treated with domestic Amsacrine (m-AMSA), singly or combined with other drugs. The total CR rate was 39.5% in ALL and 38.8% in ANLL, the response rate was 47.5% for both types of acute leukemias. The CR rate of relapsed and refractory ALL and ANLL treated with combination chemotherapy including domestic m-AMSA was 30.8% and 46.2% respectively. Domestic m-AMSA was similar to the foreign product and many other antitumor drugs in side effects and toxicity. The pharmacokinetics parameters of the drugs, C12h/C6h,K21 and Cmax were correlated with the therapeutic effectiveness. PMID- 8404330 TI - [Tumor necrosis factor and interleukin 6 in acute leukemia]. AB - In order to clarify the role of interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF alpha) in the pathogenesis of acute leukemia, IL-6 and TNF alpha level was determined in patients with various types of acute leukemia. In comparison with normal subjects, IL-6 activity was significantly elevated in patients with ALL and ANLL (P < 0.01) and TNF alpha level increased in patients with ANLL (P < 0.05). The effect of IL-6 and TNF alpha on leukemia cell in vitro was also observed. The results indicated that IL-6 can promote the proliferation of leukemia cells, while TNF alpha can inhibit proliferation of leukemia cell in vitro. It is suggested that abnormal level of TNF alpha and IL-6 in patients with acute leukemia is probably related to the pathogenesis of acute leukemia. PMID- 8404331 TI - [Study of the autologous mixed lymphocyte reaction in patients with chronic ITP]. AB - Autologous mixed lymphocyte reaction (AMLR) was studied in 37 patients with chronic ITP (including 27 active cases and 10 inactive cases) and 37 sex and age group matched healthy controls. AMLR was markedly decreased in all the patients of both active and inactive groups (P < 0.05). T cells and non-T cells from the peripheral blood of 10 patients with ITP and 10 Sex and age group matched healthy controls were investigated for HLA-DR expression by direct immunofluorescence. The percentage of DR+ non-T cells was significantly decreased in Patients with ITP as compared with the controls (P < 0.01) and there was a positive between AMLR and platelet counts (r = 0.92, P < 0.01; r = 0.75, P < 0.05 respectively). In addition, the percentage of DR+T cells was significantly increased in patients with ITP in comparison with the controls (P < 0.01). There was an inverse correlation between the percentage of DR+T cells and platelet counts (r = -0.82, P < 0.01) as well as AMLR (r = -0.81, P < 0.01). This study suggested that these were defects of the immunoregulatory function in patients with ITP as manifested by decreased AMLR, which might be related to the abnormal expression of surface DR+ antigen on non-T cells. PMID- 8404332 TI - [Increased concentration of plasma atrial natriuretic peptide in patients with acute stroke and its significance]. AB - Plasma atrial natriuretic peptide (ANP) was measured with radioimmunoassay in 39 patients with acute stroke. The mean level was 144.3 +/- 8.7 pg/ml; this was significantly higher in comparison with those in normal controls (55.8 +/- 4.7 pg/ml, n = 20) and hypertension patients (87.7 +/- 8.0 pg/ml, n = 15). Serial measurement of ANP level in 6 patients showed that it was elevated on the first day of the stroke, reached to its peak on the second or third day and returned to normal after two weeks. Five patients had hyponatremia following the attack of stroke and the plasma ANP level was further elevated. It is suggested that elevated plasma ANP level might be a pathogenetic factor of the accompanying hyponatremia in stroke. PMID- 8404333 TI - [Dynamic change of serum tumor necrosis factor activity and mechanism of hepatocyte growth factor treatment in fulminating viral hepatitis]. AB - The activity of tumor necrosis factor (TNF) in serum and in peripheral blood monocytes (PBMC) was determined in 8 patients with fulminating viral hepatitis (FVH), 10 patients with chronic active hepatitis (CAH) and 10 health controls. The activity was monitored in FVH patients before and after treatment with hepatocyte growth factor (HGF). It was found that TNF level was significantly higher in CAH patients than in controls (t = 3.56, P < 0.01) and also significantly higher in FVH than in CAH patients (t = 3.07, P < 0.01). The activity of TNF decreased gradually in FVH patients after HGF treatment and there was a positive correlation (r = 0.09, P < 0.01) between the activity of serum and PBMC TNF. It was found that in animal models of hepatic necrosis there was transient elevation of TNF activity with its peak occurring after 6 hours; the peak could be lowered with HGF treatment (t = 3.65, P < 0.05). The authors are of the opinion that TNF is an important mediator causing hepatic necrosis. The relationship between HGF and TNF was also discussed. PMID- 8404334 TI - [Classification and embolization of spinal cord arterial--venous malformation]. AB - Spinal cord arterial-venous malformation (SCAVM) is a kind of rare disease only accounting for 2%-4% of all spinal diseases. This article reports 41 cases of SCAVM out of 300 times of spinal angiography. The recent classification indicates that out of the 41 cases of SCAVM there are 26 cases of intramedullary AVM. 5 cases of perimedulla AVF. 5 cases of spinal dural AVF. 3 cases of Cobb's syndrome and 2 cases of vertebral angioma. They were treated in three ways: embolization only, embolization plus operation and operation only. As a result, the 5 paralyzed patients (3 cases of Cobb's syndrome and 2 cases of vertebral angioma) recovered totally. The factors influencing satisfactory recovery after SCAVM treatment are as follows: (1) Pure steal flow as the main cause is cured. (2) Intravertebral venous hypertension is alleviated. (3) Intravertebral occupied lesion is solved. The factors for unsatisfactory recovery after SCAVM treatment are as follows: (1) The feeding artery and parenchymal artery are embolized. (2) Drainage vein is damaged or thrombosed. (3) Intramedullary hemomyelia and spinal cord is damaged. (4) There is prolonged ischemia and spinal cord atrophy. PMID- 8404335 TI - [Super-selective angiography: analysis of angio-architecture and embolization of cerebral arterio-venous malformation]. AB - The angiograms of cerebral arterio-venous malformation (AVM) were analysed through 198 super-selective procedures. The AVM nidus as a whole can be classified as: single supply artery and multi-drainage veins; multi-supply arteries and single drainage vein; multi-supply arteries and multi-drainage veins. Angioarchitecture was divided into five types: terminal supply, penetrating supply, A-V shunt, combined with aneurysm and with anomalous venous structure. Supper-selective angiography was also used for regional functional test and A-V circulation time calculation, as a correct guidance for embolization treatment. PMID- 8404336 TI - [Embolization treatment of carotid-cavernous fistula with balloon catheter]. AB - Fifty-five cases of carotid-cavernous fistula (CCF) were treated with detachable balloon catheter from Oct. 1986 to Aug. 1992. Of these cases, 52 were embolized successfully with patented carotid arteries in 36. The clinical data, neurological complications and embolization methods were evaluated. We believe that intravascular embolization is the best method for treating CCF. PMID- 8404337 TI - [Embolization of carotid-cavernous fistulas by retrograde catheterization via superior ophthalmic vein]. AB - Six patients with traumatic carotid-cavernous fistula (CCF), which was failed to the standard endoarterial embolization for various reasons, were successfully cured by retrograde catheterization via the superior ophthalmic vein (SOV). The well dilated SOV was easily exposured (4 cases) or punctured (2 cases). Catheterization was performed under X-ray monitoring and reached into the cavernous sinus. Detachable balloon was used in 4 and IBCA glue in 2 cases for embolization. There were no intra or post-operative complication. All the symptoms and signs were disappeared in all the 6 cases after treatment. The authors suggest that in some CCF cases the SOV route is a promising approach. PMID- 8404338 TI - [Treatment of intracranial saccular aneurysms by detachable balloon]. AB - Eight cases of intracranial saccular aneurysm treated with detachable balloon were reported in this article. The aneurysms were located at ophthalmic artery (2 cases) anterior and posterior communicating artery (2 cases respectively), and basilar artery (2 cases). The detachable balloon was successfully put into the aneurysmal sac in 6 cases and obliterated the parent artery in another 2 cases. The material used to fill the balloon is made by our own hospital. All the cases were cured with the exception of 1 patient who had a contralateral hemiparesis. The indication of using detachable balloon and its technique were discussed in detail. PMID- 8404339 TI - [Long-term efficacy of endovascular embolization with silk particles and their effects on surrounding tissues in rabbits]. AB - Long-term efficacy of therapeutic embolization with silk particles in rabbits was observed. The effectiveness of embolization and their histotoxicity on surrounding tissues were compared with dry dura mater particles, polyvinyl alcohol foam (i.e. Ivalon) and isobutyl 1-2-cyanoacrylate (IBCA). The results showed that the endovascular embolization with silk particles was almost as effective as that with IBCA and superior to dry dura mater particles or Ivalon. The toxicity of silk particles was similar to Ivaion, but less than IBCA and dry dura mater particles. The authors considered that silk particles are a good embolic agent at present. PMID- 8404340 TI - [Gallbladder contractile function in patients with portal hypertension]. AB - Gallbladder fat meal-stimulated contractile function and its relation with portal hemodynamic changes in patients with portal hypertension were observed using ultrasonic pulsed Doppler duplex system. The results showed that the patients gallbladder contractile function decreased significantly and there was a negative correlation between gallbladder empty rate and portal vein diameter, congestion index and blood flow volume. It was suggested that the decreased gallbladder empty rate may be one of the causes in gallstone formation in cirrhotic patients. PMID- 8404341 TI - [Intrahepatic biliary gland proliferation its relation to acute and chronic cholangitis in hepatolithiasis]. AB - Hepatolithiasis was studied histochemically and immunohistochemically in 100 cases. The results showed that there were numerous proliferated glandular structures with in intrahepatic bile duct wall verified by consecutive sections and mucus staining and tiny abscesses in the vicinity of biliary glands in the course of acute cholangitis. It is suggested that biliary infection may occur both intramurally and extramurally. When cholangitis subsided, we observed profound lymphocytes infiltration within the glandular structures. It was concluded that biliary glands not only served as a dominant channel for the spread of acute cholangitis but also had close relation to the persistence and recurrence of cholangitis. PMID- 8404342 TI - [Autogenous vein graft as a conduit for repairing the laryngeal nerve deficit]. AB - Management of recurrent laryngeal nerve (RLN) deficit is a difficult problem and a challenge to surgeons. Effective method for repairing a RLN deficit is not available. Since October 1981, the author has adopted autogenous vein graft as a conduit to repair RLN deficit in 5 cases. All the 5 cases were followed up for more than one year after operation. The results showed that the affected vocal cord movement was entirely or nearly entirely restored in 2 cases, partially restored in 1 case, unrestored in 2 cases, patients phonation was restored to be normal or near normal in 4 cases, including 3 cases with entirely or partially restoration of vocal cord movement and one case with unrestoration of vocal cord movement. The remaining one case had vocal cord movement unrestored, the hoarseness, however was improved significantly. The data suggests that autogenous vein graft as a conduit for repairing RLN deficit is a convincible and an effective method. PMID- 8404343 TI - [The clinical application of deltoid spina scapula muscular pedicle flaps]. AB - The authors report a self-designed procedure of deltoid spina scapula muscular pedicle bone graft which is suitable for treatment of humeral bone tumor, traumatic bone defect, bone infection or anomalies involving the shoulder region and the humerus when bone graft is needed. The results of preliminary application in 11 cases show that owing to good quality and satisfactory quantity of the graft rich in vascularization, this procedure is reliable, easy to perform and facilitates rapid bone healing. PMID- 8404344 TI - [Histological and clinical characteristics of cutaneous hemangiomas and vascular malformations]. AB - One hundred and forty-two specimens of cutaneous vascular lesions were reviewed. All specimens were classified as hemangiomas (n = 114) or vascular malformations (n = 28) from this investigation. Hemangiomas in the proliferating phase (n = 55) were characterized by endothelial hyperplasia and in the regressive phase (n = 59) by fibrinoid degeneration and fat infiltration. The average number of mast cells was 25.73 +/- 9.82 in the proliferative phase and 5.12 +/- 3.24 in the regressive phase. Vascular malformations were characterized by normal endothelial turnover and a normal mast cell count (4.15 +/- 1.45). Clinically, hemangiomas exhibited a history of rapid neonatal growth and slow involution. Vascular malformations grew in proportion to the child and failed to regress. PMID- 8404345 TI - [Protein metabolism after total small bowel resection]. AB - Stable isotope 15N-lysine, 15N-alanine and 15N-glycine were used to evaluate the protein metabolism in a patient sustained by TPN for more than 4 years after total small bowel resection. Compared to normal control, the patient's total amount of absorption and urinary excretion were lower but utilization higher, the patient was considered in a status of increased protein synthesis and catabolism. Hence, the patient was vulnerable to nutritional failure if nutrition supply is not efficient. The results suggest that the nutritional program should be individualized, especially in the patient with total small bowel resection. PMID- 8404346 TI - [Application of double-J stent in treatment of pelvic-ureteric]. AB - From May 1988 to October 1991, we used double-J tube for uretero-vesical drainage as postoperative stent in 12 cases of pelvic-ureteric junction stricture. The tube was inserted either by a percutaneous antegrade procedure or through the operative site. Postoperative follow-up observation revealed satisfactory results in all cases. Compared with the traditional method, it is advantageous in providing early convalescence, preventing infection, promoting healing of the operative site or the anastomosis site, and shortening the period of hospitalization. PMID- 8404347 TI - [Experimental study on the application of biological membrane to prevent adhesions in the flexor tendon repair]. AB - Flexor digitorum profundus of adult Leghorn hens was used in the experiment. CONTROL GROUP: the long flexor tendons were cut and sutured with sheath excised between A2 and A4. Experimental group: the biological membrane was placed circumferentially around the repair site to reconstruct the flexor sheath after flexor tendon repair. The tendon healing, the adhesions and the development of pseudosheath were examined by naked eyes, optical and electron microscopy respectively. It was proved that the membrane not only could separate the healing flexor tendon from the surrounding tissue and reduce adhesion, but did not cause any local and systemic adverse reaction. The results showed that the development of the new tendon sheaths around the biological membrane was similar to silastic prosthesis in morphology. The pseudosynovial sheath supplied good tissue bed for tendon gliding. It was found that the stimulation of the membrane to the surrounding tissue was the basis of new sheath formation in the early stage and tendon gliding in the late stage would promote the pseudosheath to be mature and stable. Therefore, the results provided experimental basis for clinical practice. PMID- 8404348 TI - [Intravascular embolization in treatment of craniocerebral vascular diseases]. AB - From Aug. 1986 to June 1992, 193 cases of craniocerebral vascular diseases were treated with intravascular embolization. Among these there were 52 cases of carotid-cavernous fistula (CCF), 31 intracranial aneurysms, 83 arteriovenous malformations (AVM), and 27 dural AVMs. Intravascular embolization was first used in the treatment of CCF and dural AVM. For those of non-clippable aneurysms, detachable balloon catheter was adopted, but investigation of whole brain vessels and Matas' test should be made before embolization. As to brain AVM, embolization or combination of embolization and surgery is practical in some cases. Use of IBCA glue or silk suture in embolization is depends on the anatomical feature of AVM. PMID- 8404349 TI - [Determination of retinal illumination from operating microscopes and assessment of risk]. AB - The retinal illumination from 5 popular operating microscopes was determined with the authors' method to reveal that the intensity of light differed very significantly between brightness adjustments. With dilated pupil and clear media, the power of illumination on the retina was 0.072-0.233 W/cm2 at maximum brightness and 0.042-0.084 W/cm2 at medium brightness, and the safety time limits for application were 22.49s for the former and 50.38s for the latter to avoid retinal damages. PMID- 8404350 TI - [A molecular genetic study of Leber's disease]. AB - That Leber's hereditary optic neuropathy is caused by a single nucleotide change in the mitochondrial DNA (mtDNA) was first verified by Wallace and his colleagues in 1988. The mtDNA of 11 patients with Leber's disease from different families and of normal persons are analyzed with polymerase chain reaction by the authors, with results that confirm those of Wallace. The procedure is simple and speedy, and hence recommended for clinical utilization. PMID- 8404351 TI - [The C/D ratio in normal newborns]. AB - The optic disc and the C/D ratio were examined in 122 newborns to find that the features of the C/D ratio were positive skewness distribution with leptokurtosis, a wide variability, lesser values of the mean, the median and the 95% upper limit, and non-correlation with the length of pregnancy and the height and weight of the baby at birth. The authors suggested that C/D > 0.3 or a binocular difference of > 0.2 could be regarded as one of the criteria of newborn glaucoma. It was also found that more greyish glial tissue was on the optic disc. The scope of the physiological optic cup was determined by the naked status of the central retinal vessels in the pale area. PMID- 8404352 TI - [Investigations on the latent infection of herpes simplex virus-1 in the cornea]. AB - The relapsing rate in 216 patients of recurrent herpes simplex keratitis (HSK) treated with penetrating keratoplasty was 4.2% in 6.5 years, while experimental studies showed that HSV-1 antigens in the cornea disappeared 45 days after the primary corneal infection. Among 18 cases of quiescent HSK, HSV-1 antigens were negative in the corneal tissue, which however turned positive in 6 (66.7%) of 9 cases that were cultured for reactivation, and became infectious to primary rabbit kidney cells. Tests of susceptibility revealed that rabbit keratocytes were most sensitive to infection with HSV-1 strain Mckrae. The authors opined that the cornea might be an additional site to the trigeminal ganglion to harbor HSV-1. PMID- 8404353 TI - [The effect of fibronectin on re-epithelialization of rabbit cornea after alkali burn]. AB - The authors found experimentally that (1) fibronectin enhanced the healing of rabbit corneal epithelium after alkali burn and prevented the secondary breakdown; (2) it rapidly deposited on the denuded basement membrane to disappear as epithelial cells slided over, and (3) ultrastructurally, the neighbouring epithelial cells became flattened, with filopodia at the advancing edge, and extended to the wound area 24 hours after the burn. However, the epithelial defect recurred 72 hours after the burn when the basement membrane vanished till after the 8th week, indicating that the latter was associated with the secondary epithelial breakdown. PMID- 8404354 TI - [Stages of primary pupil-block chronic angle-closure glaucoma]. AB - In China, primary pupil-block chronic angle-closure glaucoma is a blinding eye disease of high incidence in primary glaucoma, with different clinical course and features from acute angle-closure glaucoma. Based on the conditions of the anterior chamber angle, the level of IOP, the defects in optic disc and visual field, and the mechanism of IOP elevation, this morbid entity was classified into 5 stages, i.e., the preclinical, the early, the progressive, the advanced and the absolute stages, and the clinical manifestations of each stage were described. This classification may be of referential use to ophthalmologists in clinical and research work. PMID- 8404355 TI - [Two new types of keratoscope and their clinical application]. AB - The author reports the clinical application of his 2 new types of keratoscope for qualitative observation of the corneal curvature: the reflective type and the transparent type. The latter was particularly useful in controlling the corneal astigmatism during intraocular microsurgery. According to practical use on 30 patients, the positive rate of astigmatic detection and the recognition of astigmatic meridian by these keratoscopes were satisfactory. PMID- 8404356 TI - [Dynamic observations of the optic cup and its modes of damages in glaucoma]. AB - Changes in the optic disk and retinal nerve fiber layer are the sole objective indicators of glaucomatous damages. Many investigators held that damages in the optic nerves in glaucoma preceded changes in the visual functions. The optic cups and visual fields of 106 primary open angle glaucoma eyes and 207 eyes of glaucoma suspects were followed up an average 16.6 months. Among 35 eyes that developed changes in the optic cup, the cup changes preceded the visual field changes in 78%, and both changes occurred simultaneously in 22%. PMID- 8404357 TI - [Microcomputer image analysis of surface parameters of the optic disk]. AB - 103 normal and 86 open-angle glaucoma optic disks were evaluated with the microcomputer image analyser. Except for the diameter and area, all other parameters of the normal disk differed significantly from those of the glaucomatous disk. The cup/rim area ratio in glaucoma was significantly larger, while the cup/disk area ratio was moderately so, indicating that the former was more useful in the early diagnosis of glaucoma. The microcomputer image analyser was reliable and accurate, also capable of processing blurred images from varying exposure and focussing. The quotient between the horizontal and vertical C/D ratios was a relatively stable value with changing glaucomatous cupping. PMID- 8404358 TI - [Clinical features of primary chronic angle-closure glaucoma]. AB - 43 cases (86 eyes) of primary chronic angle-closure glaucoma (PCACG) and 44 cases (77 eyes) of primary acute angle-closure glaucoma (PAACG) randomly selected, and 30 normal subjects (34 eyes) were performed ultrasonic biometry of anterior chamber depth, lens thickness and axial length of the globe. By a micrometer attached to the slip-lamp, the width of the anterior chamber angle entrance was also calculated. The clinical manifestations and the natural course of PCACG, including the morphology of the anterior chamber angle and the peripheral anterior synechiae (PAS) as well as the dynamic changes in aqueous outflow were investigated. It was found that there were significant differences in the biometric parameters of the anterior segment between PCACG and PAACG or normal eyes. Follow-up study of the early stage PCACG cases showed that topical miotics and/or peripheral iridectomy effectively prevented the formation of PAS and the progressive development of PCACG. The criteria for diagnosis and the principles for treatment of PCACG were stipulated. The pathogenesis of PCACG was discussed, and it was emphasized that minor attacks of PAACG and its progressive stage should not be confused with PCACG. PMID- 8404359 TI - [Prevention of IOP rise following laser iridectomy with topical timolol and indomethacin]. AB - Double-masked study on 40 eyes (40 patients) of primary angle-closure glaucoma demonstrated that topical timolol and indomethacin prevented the IOP rise following combined argon and Nd:YAG laser iridectomy. A combination of the 2 drugs was especially effective in that the number of patients who would have an IOP increase over 1.33 kPa (1 kPa = 7.5 mmHg) above the baseline was reduced by 80%. The mechanism of IOP rise following iridectomy and its prevention were discussed. PMID- 8404360 TI - [The anti-cicatrization effect of low dosage 5-Fu after trabeculectomy in late glaucoma]. AB - 54 patients (66 eyes) of late primary glaucoma were divided into 5-Fluorouracil and control groups before trabeculectomy surgery. The 5-Fu group received subconjunctival injections of 5-Fu 3mg postoperatively and on each of the 6 ensuing days. An average follow-up of 14 months revealed that (1) functional filtering blebs in the 5-Fu group and the control group were 100% and 60% respectively; (2) the postoperative mean diurnal peak IOP was 16.3 +/- 4.9 mmHg in the 5-Fu group and 20.1 +/- 5.2 mmHg in the control group (P < 0.01), and eyes with diurnal IOP < 21 mmHg were 90% in the 5-Fu group and 26% in the control group at the end of the second postoperative year; and (3) the postoperative complications between the 2 groups were comparable, except for shortterm corneal defects in 7 eyes (21.2%) of the 5-Fu group. PMID- 8404361 TI - [Transplantation of functional trabecular meshwork in 9 cases]. AB - On the basis of experiments on rabbits, transplantation of functional trabecular meshwork was performed in 9 cases of absolute angle-closure glaucoma. The surgical reactions were not serious, and the postoperative intraocular pressure decreased significantly by 50%-80% on discharge from the hospital, and in 4 cases followed up for 3 months, by 20%-60%, with 1 case of ocular hypotension. This modality of trabeculoplasty opens a new prospect of ophthalmological organ transplantation for probation. PMID- 8404362 TI - [Modified threshold Amsler grid testing]. AB - The conventional Amsler grid uses suprathreshold illumination, and hence may fail to detect early scotomas. When the grid is viewed through cross-polarizing filters to reduce luminance to visual threshold, the test becomes far more sensitive. The authors expanded the standard Amsler grid from 400 to 1600 squares and viewed under threshold luminance to examine the visual fields of glaucoma, retinitis, optic neuropathies and brain tumors; the modification was found much more sensitive for the detection of central and paracentral light scotomas. PMID- 8404363 TI - [Arcuate keratotomy for the treatment of corneal astigmatism after intraocular lens implantation]. AB - Arcuate keratotomy was performed for 21 eyes of persistent corneal astigmatism > 2.25D over 6 months after intraocular lens implantation. The mean corneal astigmatism 1 day and 6 months after the procedure was 0.82D and 1.18D respectively, and the visual acuity improved significantly. The indications, manipulations, efficacy, and complications of the operation were discussed. PMID- 8404364 TI - White blood cell differentiation using a solid state flow cytometer. AB - A flow cytometer using a solid state light source and detector was designed and built. For illumination of the sample stream two types of diode lasers (670 nm and 780 nm) were tested in a set-up designed to differentiate human leukocytes by means of light scattering. The detector is an avalanche photodiode, which was used to detect the weak scattered light in the orthogonal direction. The new flow cytometer set-up is very small, relatively cheap and yields similar results as a standard flow cytometer set-up using a helium-neon laser and photomultipliers. PMID- 8404365 TI - Increased membrane permeability of apoptotic thymocytes: a flow cytometric study. AB - We have recently developed a method for the separation and quantification of viable apoptotic cells without the need for permeabilisation or fixation of the cells. The method is based on the observation that apoptotic rat thymocytes fluoresce more brightly than normal cells after a brief incubation with the DNA binding dye, Hoechst 33342. In order to understand these differences, we have investigated the uptake of Hoechst 33342 by normal and apoptotic thymocytes. By staining with fluorescein diacetate, we have shown that the efflux of fluorescein from apoptotic cells is more rapid than that from normal thymocytes. This result demonstrated an increase in the permeability of the plasma membrane of the apoptotic thymocytes and it is this change which probably results in the more rapid uptake of Hoechst 33342. The data also revealed two populations of apoptotic thymocytes. PMID- 8404366 TI - The use of flow cytometry for the investigation of cell death. AB - Flow cytometry is more and more widely used for investigations of cell death, predominantly in the study of DNA degradation in cells dying by apoptosis. There are different interpretations of changes observed in DNA histograms of these cells. We describe an approach based on extraction of chromatin degradation products from fixed cells and subsequent staining with DNA specific dyes. Apoptotic cells containing fragmented DNA are observed in < 2C DNA region of DNA histograms. DNA histograms of irradiated thymocytes dying in vitro and stained without extraction of fragmented DNA do not differ from control. Under the same staining conditions DNA histograms of lymphocytes dying in thymus of irradiated animals reveal fluorescent material in < 2C DNA region, most likely due to formation of apoptotic bodies (cell fragments, some of them contain fragments of nuclei). Similar changes are observed in thymocytes dying upon glucocorticoid treatment. Our present results and other data indicate that reduced amount of DNA in dying cells is the main reason for changes of DNA histograms. Examples of application of the method described for the investigations of cell death modifiers are presented. PMID- 8404367 TI - Equilibrium binding of DAPI and 7-aminoactinomycin D to chromatin of cultured cells. AB - The binding of 4',6-diamidino-2-phenylindole (DAPI) and 7-aminoactinomycin D (7 AAMD) to chromatin of intact cultured fibroblast nuclei was studied by cytofluorometry. Staining was performed at equilibrium at varying dye concentrations. By using large volumes of dye solutions, the free dye concentration became approximately equal to total dye concentration, and the estimation of affinity and number of binding sites was performed by Scatchard analysis. Since all Scatchard plots showed a definite curvature, indicating more than one class of binding sites, a two component regression was performed, although only the highest affinity binding sites were determined. The observed affinity constants did not change substantially between different preparational procedures for any of the dyes. The number of DAPI binding sites increased about 30% after detergent extraction. No such increase was observed for 7-AAMD. Fixation in formaldehyde, instead of ethanol, after detergent extraction had no effect on DAPI binding, but decreased the number of 7-AAMD binding sites with about 50%. Extraction of basic proteins with HCl resulted in about 100% increase of the number of binding sites for both dyes. It should be feasible to use this technique as an assay for the identification of proteins or other nuclear components important for the maintenance of chromatin structure. PMID- 8404368 TI - Heterochromatin study demonstrating the non-linearity of fluorometry useful for calculating genomic base composition. AB - A novel procedure for calculating base-pair frequencies in whole genomes is reported. This has been developed during a study of the role of heterochromatin in microevolution. Closely related species of the Crepis praemorsa complex have similar karyotypes but for their heterochromatin. The changes in relative AT frequency between species have been attributed to heterochromatin sequences by in situ banding of chromosomes with two base-specific fluorochromes. The absolute genome size of species, measured by cytofluorometry, correlated positively with increased karyotypic heterochromatin, as did the proportion of AT bases in the DNA. However, the determination of base content has called for a curvilinear interpretation of data obtained with two base-specific fluorochromes (bisbenzimide Hoechst 33342 and mithramycin), in contrast to the commonly assumed but erroneous direct relationship between fluorescence intensity and base content. Essentially, the fluorochromes' requirements for a sequence of certain base-pairs lead to the notion of Coefficients of Overspecificity: the result is a simple formula for calculating the AT proportion in a genome relative to a reference species from cytometric data, taking account of ligand binding statistics. These statistics and probabilities of oligonucleotide binding are essentially the same. PMID- 8404369 TI - Application of artificial neural networks to chromosome classification. AB - This work presents an approach to the automatic classification of metaphase chromosomes using a multilayer perceptron neural network. Representation of the banding patterns by intuitively defined features is avoided. The inputs to the network are the chromosome size and centromeric index and a coarsely quantized representation of the chromosome banding profile. We demonstrate that following a fairly mechanical training procedure, the classification performance of the network compares favourably with a well-developed parametric classifier. The sensitivity of the network performance to variation in network parameters is investigated, and we show that a gain in efficiency is obtainable by an appropriate decomposition of the network. We discuss the flexibility of the classifier developed, its potential for enhancement, and how it may be adapted to suit the needs of current trends in karyotyping. PMID- 8404370 TI - Detection of 5-bromo-2-deoxyuridine (BrdUrd) incorporation with monoclonal anti BrdUrd antibody after deoxyribonuclease treatment. AB - We studied the effects of deoxyribonucleases on the detection of 5-bromo-2 deoxyuridine (BrdUrd) by anti-BrdUrd monoclonal antibodies (mAbs). After DNase I treatment, BrdUrd was detected in cells fixed on slides with the anti-BrdUrd mAbs, B44 and BMC9318. The level of detection related to the degree of DNA digestion. DNA digestion of 25-75% resulted in levels of staining comparable to control preparations in which DNA was denatured by heating with formamide. Staining with the mAbs of DNase I-treated cells was abolished with S1 nuclease, a single-stranded DNA-specific nuclease. When exonuclease III was used after DNase I treatment, the staining intensity of cells fixed on slides increased, and BrdUrd could be detected in suspended cells by flow cytometry. Since this enzymatic method leading to the detection of BrdUrd does not involve cell loss, or destruction of either cellular morphology or epitope reactivity, as occurs with traditional DNA denaturation procedures, it is useful for kinetic studies of phenotypically mixed populations. Furthermore, staining with anti-BrdUrd mAb of cells treated with exonuclease III offers a simple approach to quantitation of apoptotic cells, in which an endogenous endonuclease is activated. PMID- 8404371 TI - Cluster analysis of flow cytometric list mode data on a personal computer. AB - A cluster analysis algorithm, dedicated to analysis of flow cytometric data is described. The algorithm is written in Pascal and implemented on an MS-DOS personal computer. It uses k-means, initialized with a large number of seed points, followed by a modified nearest neighbor technique to reduce the large number of subclusters. Thus we combine the advantage of the k-means (speed) with that of the nearest neighbor technique (accuracy). In order to achieve a rapid analysis, no complex data transformations such as principal components analysis were used. Results of the cluster analysis on both real and artificial flow cytometric data are presented and discussed. The results show that it is possible to get very good cluster analysis partitions, which compare favorably with manually gated analysis in both time and in reliability, using a personal computer. PMID- 8404372 TI - Pure populations of Dictyostelium discoideum prespore and prestalk cells obtained by flow cytometry have different redevelopment characteristics at their cell surfaces. AB - The multicellular slug stage of Dictyostelium discoideum consists of two major differentiated cell types: prespore and prestalk cells, which become, respectively, the spores and the stalk of the fruiting body. It is known that these cells, although expressing cell-type-specific proteins, remain totipotent, and experimental disruption of slugs results in redifferentiation taking place. We looked at what happens to cell-type-specific surface molecules when a cell changes from one type of another. Using monoclonal antibodies and flow cytometry we were able for the first time to obtain pure populations of single cells of each cell type. These were analysed during redevelopment. The initial hypothesis was that a proportion of each cell type would redifferentiate to reestablish the original proportions. However, it was found that the two cell types responded quite differently. Whereas almost all prestalk cells retained their prestalk surface antigen, in contrast, all prespore cells redifferentiated. During this process redifferentiating prespore cells simultaneously expressed surface determinants of both cell types, an event not seen in normal development. PMID- 8404373 TI - False aneuploidy in flow cytometric DNA analysis of paraffin embedded tissue: effects of Carnoy's fixation. AB - False aneuploidy was detected on flow cytometric DNA analysis of paraffin embedded axillary lymph nodes negative for tumor. It was hypothesized that "clearing" of axillary fat in Carnoy's solution to facilitate lymph node dissection might be responsible for false aneuploidy. Various tissues fixed overnight in Carnoy's were compared to formalin fixed paraffin embedded controls. Under these conditions no false aneuploid peaks were detected, but Carnoy's fixation did shift the G0/G1 histogram peak to the left, increase the G0/G1 CV and increase the S phase fraction relative to formalin fixed controls. It was then hypothesized that partial fixation of nodes in Carnoy's followed by formalin fixation might result in false aneuploid peaks. Twenty-two lymph nodes were partially fixed in Carnoy's for periods ranging from 5 to 60 min followed by complete fixation in formalin. Seven of these nodes did show false aneuploid peaks. By contrast, no aneuploidy was detected in formalin fixed controls. It was concluded that tissues in contact with Carnoy's solution may be a source of false aneuploidy and/or false elevation of S phase fraction. This reinforces the need for matched negative tissue controls for DNA analysis of paraffin embedded specimens whenever possible. PMID- 8404374 TI - Improved DNA content histograms from formalin-fixed, paraffin-embedded liver tissue by proteinase K digestion. AB - An improved method for the enzymatic digestion of formalin-fixed, paraffin embedded liver tissue for DNA content analysis by flow cytometry is presented. Forty samples of histologically normal liver were alternately digested by the traditional pepsin method or a new method utilizing proteinase K and heat. Sixteen (40%) of the pepsin-digested samples had apparent DNA aneuploid peaks by flow cytometry. False DNA aneuploid peaks were not present in any of the histograms obtained after proteinase K digestion. Microscopy showed that the pepsin-digested samples had residual cytoplasmic remnants which contained fluorescent material. Samples digested with proteinase K had few cytoplasmic remnants. The average G0/G1 coefficient of variation after proteinase K treatment was lower (41%) and the fluorescent intensity higher (128%) than the pepsin treated samples. The apparent mean S-phase (a combination of S-phase cells and underlying debris) after proteinase K digestion was 35% of the pepsin-treated samples. Primary and secondary tumors of the liver that were DNA aneuploid after pepsin treatment were also DNA aneuploid after proteinase K treatment. A modified digestion protocol utilizing proteinase K and heat can provide superior results for DNA content analysis of formalin-fixed, paraffin-embedded liver tissue. PMID- 8404375 TI - Double-staining artefact observed in certain individuals during dual-colour immunophenotyping of lymphocytes by flow cytometry. AB - An unusual double-staining artefact has been repeatedly observed in certain individuals (both HIV-positive and healthy laboratory controls) during 2-colour immunophenotyping of lymphocytes by flow cytometry after red cell lysis. This artefact can falsely suggest co-expression of CD4 and CD8, as well as some of the other monoclonal antibody pairs commonly used in the typing of lymphocytes. It is caused by a serum factor associated with the ammonium-sulphate-precipitated and gamma-globulin fractions. It appears to be fairly common in the population (being found in 6 out of 105 of our healthy laboratory controls and in 42 out of 247 of our HIV-1 seropositive subjects); it may be genetically determined, but may be acquired in some. It is a potential source of error in lymphocyte immunophenotyping, which can be avoided by simple adjustments to the recommended techniques. PMID- 8404376 TI - Determination of the percentage of thiazole orange (TO)-positive, "reticulated" platelets using autologous erythrocyte TO fluorescence as an internal standard. AB - Determination of the percentage of thiazole orange (TO)-positive or "reticulated" platelets by flow cytometry has been advocated as an aid in the diagnosis of thrombocytopenic disorders. However, a reproducible method for determining control fluorescence and setting threshold values on a routine clinical basis has not been described. We used erythrocyte TO fluorescence in whole blood as an internal standard to set threshold markers for TO fluorescence of autologous, purified platelets. Since platelets have approximately a threefold higher TO fluorescence than erythrocytes, multiplying the erythrocyte threshold marker by 3 allowed a determination of the percentage of "reticulated" platelets in 29 normal controls. This value was 3.64% +/- 2.10% (S.D.) in men and 5.79 +/- 2.22% (S.D.) in women; there was a significant (P < 0.05) difference between normal men and women for this measurement. Applying this method to our earlier studies of normals and patients with immune thrombocytopenic purpura confirmed its ability to discriminate between these populations with similar sensitivity and specificity as previously reported. This method controls for fluctuations in TO labeling and fluorescence and does not require daily determination of a new normal control value. PMID- 8404377 TI - Flow cytometric analysis of oxygen species formation in activated leukocytes. PMID- 8404378 TI - Predicting the functional result of anastomoses to the anus: the paradox of preoperative anal resting pressure. AB - This article examines the effect of ileal pouch-anal (n = 134) and coloanal (n = 16) anastomoses on resting anal canal pressures in 150 patients. METHODS: Patients underwent anal manometry before ileal pouch-anal anastomosis (IPAA) and coloanal anastomosis (CAA) and again six weeks after ileostomy closure following these procedures. A water-perfused catheter system with four radial ports was used for manometry, pressures being recorded during both station and continuous pull through. RESULTS: Patients with IPAA were younger than those with CAA (34 years vs. 50 years) and had a different ratio of hand-to-stapled anastomosis (1:2.6 vs. 1.3:1). All CAA patients had had rectal cancer while IPAA patients suffered mainly from ulcerative colitis (n = 114) or familial polyposis (n = 10). The mean preoperative resting pressure for all patients was 79 mmHg (75-87, 95 percent confidence limit) and the mean fall in this pressure after surgery was 25 mmHg (-21 to -29, 95 percent confidence limit). There was no difference in preoperative pressure or fall between handsewn and stapled anastomoses, or between IPAA and CAA. CONCLUSION: There was a significant relationship between preoperative pressure and change in pressure that held true for all subgroups (change = -0.7 x preoperative pressure + 31, r = 0.69). Analysis of the functional results confirmed that patients with high preoperative pressure are at risk for severe falls after surgery and are not guaranteed a good result. Conversely, patients with low preoperative pressures may actually have an increase with surgery and are not always incontinent. Patients with low preoperative anal resting pressures should not be denied anastomosis to the anus if they are continent. PMID- 8404379 TI - Dynamic anal manometry in the assessment of patients with obstructed defecation. AB - Patients with obstructed defecation show no consistent abnormalities when assessed by standard anorectal physiologic methods. With a recently developed technique for dynamic anal manometry, we studied 13 female patients with obstructed defecation and 20 healthy volunteers. Seven parameters of anal function were measured. There were no differences between the median values for the two groups. Seven patients (54 percent; 95 percent confidence limits, 25-81 percent) had anal compliance below the normal range, either during opening or closing of the sphincter at rest (five patients), during squeeze (one patient), or both (one patient). Opening and closing pressures of the sphincter at rest, maximal closing pressure during squeeze, and anal hysteresis were normal. Standard anal manometry did not show any differences between patients and controls. Rectal compliance was lower in patients with obstructed defecation, median difference 5 ml/cm H2O (95 percent confidence limits, 1-9 ml/cm H2O). In conclusion, the more detailed method of dynamic anal manometry shows that some patients with obstructed defecation have a less compliant anal sphincter and a less compliant rectum, but in many patients no abnormal findings can be made. PMID- 8404380 TI - Increased risk of early colorectal neoplasms after hepatic transplant in patients with inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is associated with an increase in colon and rectal carcinoma. Immunosuppression after transplantation increases the incidence of certain types of tumors. PURPOSE: We reviewed the postoperative course of IBD patients who had undergone hepatic transplantation for primary sclerosing cholangitis to see whether there was an increase in the rate of colorectal neoplasms. METHODS: The charts of 44 patients from two institutions who had undergone a hepatic transplant for primary sclerosing cholangitis were reviewed. Of these 44 patients, 33 had IBD (32 chronic ulcerative colitis, 1 Crohn's). Of these 33 patients, 2 had previously undergone total colectomy/proctectomy and 4 died in the perioperative period. The remaining 27 patients had all undergone colonoscopic evaluation just prior to transplant. Postoperatively all patients were given prednisone, cyclosporine, and azathioprine. Minimum follow-up was 12 months; mean follow-up was 39 months. RESULTS: Three of the 27 patients (11.1 percent) developed early colorectal neoplasms (2 cancers, 1 large villous adenoma with severe dysplasia) at 9, 12, and 13 months post-transplant. All three patients were successfully treated with a total colectomy/proctectomy or resection of any remaining colon. These 3 patients had a mean 19-year history of IBD (range, 9-27 years), while the 24 patients without tumors had a mean 18-year history of IBD (range, 6-39 years). CONCLUSION: There is a subset of transplant patients with primary sclerosing cholangitis and IBD who rapidly develop colorectal neoplasms. Frequent surveillance is recommended for IBD patients in the post-transplant period. PMID- 8404381 TI - Decreased levels of protein kinase C enzyme activity and protein kinase C mRNA in primary colon tumors. AB - PURPOSE: We have previously reported decreased protein kinase C (PKC) enzyme activity in primary human colorectal carcinomas. The purpose of this study was to extend these findings to a larger number of cases and to also examine the levels of expression of mRNAs that encode specific isoforms of PKC in these tumors. METHODS: Colorectal carcinomas and paired grossly normal adjacent mucosal samples were collected from 39 patients. Complete histopathologic analyses were performed on all samples. PKC enzyme activity in both the cytosolic and particulate fractions was quantitated by measuring the amount of 32P incorporated into histone Type III-S. Northern blot nucleic acid hybridization was performed using polyA+ RNA extracted from both the tumor and normal tissue samples and 32P labeled probes for specific isoforms of PKC. The paired sample t-test was used to determine the statistical significance of tumor to normal ratios of both enzyme activity and mRNA levels. RESULTS: The mean value for cellular PKC enzyme activity in the colon tumors from 39 patients was about 60 percent of that found in the paired adjacent grossly normal mucosa samples (P < 0.001). The subcellular distribution of PKC activity was similar in normal and tumor samples (about 70 percent in the particulate fraction). The abundance of PKC alpha mRNAs varied considerably among 28 tumor/normal pairs, with a mean tumor to normal (T:N) ratio of 1.0 +/- 0.6 for the 9.9-kb mRNA band and 1.4 +/- 0.7 for the 3.5-kb band. The abundance of PKC beta mRNAs was decreased in 30 of 39 tumors, with a mean T:N ratio of 0.6 +/- 0.4 for both the 9.4- and 3.5-kb bands for all 39 samples (P < 0.001). None of the parameters measured correlated with Dukes stage or the grade of the tumor. CONCLUSIONS: These studies extend previous evidence that total PKC enzyme activity is frequently decreased in primary human colon tumors. Our finding that this is often associated with decreased levels of PKC beta mRNA suggest that this is not simply due to post-translational down-regulation of this enzyme system. Further studies are required to determine whether these changes in PKC alpha and PKC beta mRNAs are due to altered de novo transcription or mRNA stability. It will also be of interest to examine the expression of other isoforms of PKC in colon tumors. PMID- 8404382 TI - Alteration in intestinal permeability after ileal pouch-anal anastomosis. AB - PURPOSE: The physiologic changes that occur when the small bowel is used as a reservoir, as in the ileal pouch-anal anastomosis, are poorly understood. Alterations in bowel permeability, which may lead to bacterial translocation that could result in illness or dysfunction of the pouch, may be one such consequence of the pouch procedure. METHODS: Whole-bowel permeability was evaluated in patients with and without the pouch through the use of an orally consumed nonmetabolizable sugar clearance technique. Patients in whom the ileal pouch-anal anastomosis was performed for ulcerative colitis (17 patients) and patients with familial polyposis (7 patients) were compared with normal healthy volunteers (10 patients) and patients with ulcerative colitis with and without curative colectomy and ileostomy (6 and 5 patients, respectively). RESULTS: Measured by this technique, no differences were noted in bowel permeability between the volunteers and patients with ulcerative colitis, even after colectomy and ileostomy (1.7 +/- 0.4 in normal healthy volunteers, 1.8 +/- 0.5 in patients with ulcerative colitis without stoma, and 1.4 +/- 0.2 in patients with ulcerative colitis with ileostomy). The group of patients with an ileal reservoir, however, had a significantly increased index of measured bowel permeability (3.5 +/- 0.5 in patients with ulcerative colitis and 5.1 +/- 0.7 in patients with familial polyposis; P < 0.05 by analysis of variance compared with normal healthy volunteers and patients with ulcerative colitis with or without ileostomy). CONCLUSION: The exact site, cause, and consequence of this possible alteration of bowel permeability are unclear but appear to be related to the presence of the pouch and are not caused by the underlying pathologic diagnosis. PMID- 8404383 TI - Treatment and follow-up strategies in hereditary nonpolyposis colorectal carcinoma. AB - The treatment and follow-up strategies of patients with hereditary nonpolyposis colorectal carcinoma (HNPCC) were analyzed in 22 Finnish HNPCC families in systematic follow-up between 1983 and 1990. During the seven-year study period metachronous colorectal neoplasia was diagnosed in 41 percent (15/37) of the patients treated by segmental colonic resection and in 24 percent (4/17) of those treated by subtotal colectomy. Extracolonic carcinoma was diagnosed in 12 (30 percent) of the 40 patients during the long-term follow-up. The most common extracolonic malignancy was biliopancreatic carcinoma which accounted for all five cancer-related deaths in the whole series during the study period. It was concluded that subtotal colectomy is superior to hemicolectomy or segmental resection in HNPCC patients with colorectal carcinoma. A regular annual endoscopic follow-up of the residual rectum is still necessary, and surveillance for extracolonic cancers must be considered. PMID- 8404384 TI - Anticarcinoembryonic antigen immunoscintigraphy with a 99mTc-Fab' fragment (Immu 4) in primary and recurrent colorectal cancer. A prospective study. AB - Forty-seven patients were submitted to 68 radioimmunoscintigraphic investigations for primary or recurrent colorectal cancer. Immunoscintigraphy with Immu-4 correctly detected 28 primary colorectal cancers of 29 and 12 of 12 recurrent colorectal cancers. Overall accuracy was 93.75 percent in primary and 91.6 percent in recurrent colorectal cancer. Immunoscintigraphy had a decisive influence on treatment planning in every third primary colorectal cancer patient and was by far superior to CT scan in the detection of early recurrences, especially in patients with a history of abdominoperineal or low anterior resection. Immu-4 scintigraphy is a safe and convenient diagnostic approach to colorectal cancer. Because radioactivity is acceptably low and the method is absolutely free of side effects, there are no objections to the repeated use of immunoscintigraphy which provides important information in primary diagnosis as well as in the follow-up of colorectal cancer patients. PMID- 8404385 TI - Pelvic abscess after colon and rectal surgery--what is optimal management? AB - PURPOSE: The aim of this study was to compare treatment outcomes in the management of pelvic abscess (PA) after rectal surgery. METHODS: Over a 12-year period all PAs occurring in the patients undergoing colorectal resection were retrospectively reviewed. The APACHE II Score was used to stratify illness. RESULTS: Postoperative PA developed in 56 patients after cancer (32 percent), ulcerative colitis (26 percent), diverticular disease (24 percent), and Crohn's colitis (18 percent)/surgery. Overall, 24 (43 percent) of PAs were after operations for inflammatory bowel disease and 43 (77 percent) of PAs were after intrapelvic intestinal anastomoses. PAs were treated by 1) antibiotics alone (11/56), 2) percutaneous computerized tomography-guided catheter drainage (13/56), 3) transperineal drainage (15/56), or 4) laparotomy (17/56). Recurrent PAs developed in 11/56 (19 percent) after initial treatment, of which 7 required additional surgery. These recurrences were evenly distributed between treatment groups. There were three deaths as a result of PA, two after laparotomy and one after percutaneous drainage. Long-term sequela in patients with intestinal anastomosis included loss of intestinal continuity (10/43) and anastomotic stenosis (7/43). There was no difference in APACHE II Score among the four treatment groups. The mortality rate was 75 percent among patients whose APACHE II Scores were greater than 15. The development of a PA after colon and rectal surgery was associated with a 5 percent mortality and 41 percent functional morbidity (23 percent permanent stoma and 18 percent symptomatic stricture rate). CONCLUSION: Using clinical judgment, if PA is amenable to computerized tomography guided percutaneous or transperineal drainage, one of these techniques should be attempted initially in the hemodynamically stable nonseptic patient. Long-term functional disability is common after PA in rectosigmoid surgery in patients who undergo pelvic/intestinal anastomosis. PMID- 8404386 TI - Proctoscopic Doppler ultrasound in diagnostics and treatment of bleeding hemorrhoids. AB - PURPOSE: Endoscopic Doppler sonography is a relatively new technique in the diagnostics of intestinal hemorrhage. It has been used mainly for bleeding gastroduodenal ulcers, but can also be utilized in the lower digestive tract. METHODS: In the study presented 80 patients with symptomatic hemorrhoids of first degree and previous hemorrhage were randomized in two groups. The 40 patients in Group A were examined by transproctoscopic Doppler ultrasound. After measuring the vessels depth, local injection treatment with 5 percent phenol almond oil followed. The patients in Group B had been given conventional sclerotherapy without the aid of Doppler investigation. The success of treatment was checked using Doppler sonographics six weeks later and the results were statistically compared. RESULTS: In the patients in Group A, the hemorrhoids proved to be totally eliminated in 93 percent vs. 38 percent of the patients in Group B (P, 0.1 percent). CONCLUSIONS: Proctoscopic Doppler ultrasound is useful in the evaluation and sclerotherapy of symptomatic first-degree hemorrhoids. PMID- 8404387 TI - A new probe for measuring electromyographic activity from multiple sites in the anal canal. AB - A new multiple electrode probe (MEP) designed to measure surface electromyographic activity from the subcutaneous and deep portions of the external anal sphincter is described. Using the MEP, electromyographic activity was sampled in an asymptomatic subject and in three incontinent patients. Comparisons of electromyographic data between and within subjects, and across sessions, indicate that the MEP discriminates muscle activity from different sites along the anal canal. The recording method described is capable of identifying patterns of muscle recruitment which have not been reported before using surface electromyography. After a defecation maneuver, the distal and proximal portions of external and sphincter were observed to contract in what would seem to be a closing reflex. Moreover, the absence of this pattern may indicate abnormality. Accordingly, the MEP promises to be useful in the diagnosis, evaluation, and treatment of fecal and urinary incontinence and, particularly, in the biofeedback treatment of these and related disorders. PMID- 8404388 TI - Neoplastic transformation arising in Peutz-Jeghers polyposis. AB - PURPOSE AND METHODS: To clarify the potential for malignancy of Peutz-Jeghers polyposis, we investigated 75 gastrointestinal polyps resected surgically or endoscopically from seven patients with this syndrome. RESULTS: There were 19 polyps in the stomach, 18 in the duodenum, 22 in the small intestine, and 16 in the large intestine, and these were histologically composed of 1 pyogenic granuloma, 1 cancer in adenoma, 2 adenomas, and 71 Peutz-Jeghers polyps. Nine of these Peutz-Jeghers polyps were accompanied by an adenomatous component, and, in addition, two of these showed a cancerous transformation with stalk invasion. A total of 12 neoplastic polyps (16 percent) were found in three relatively young patients (aged 20, 25, and 43 years), all of which were pedunculated and located either in the duodenum or in the jejunum. There was no statistical significance in size between the neoplastic polyps (mean +/- SD, 20.1 +/- 10.8 mm) and the completely hamartomatous polyps (mean +/- SD, 15.8 +/- 9.0 mm). Moreover, the configuration of these types of polyps seemed similar. CONCLUSION: Neoplastic transformation is not a rare event, and our results may indicate evidence of a hamartoma-adenoma-carcinoma sequence in Peutz-Jeghers polyposis. PMID- 8404389 TI - Long-term results of large-dose, single-session phenol injection sclerotherapy for hemorrhoids. AB - PURPOSE: The aim of our study was to assess the medium to long-term outcome following single session large dose injection sclerotherapy for symptomatic hemorrhoids. METHODS: One hundred eighty-nine patients (male = 106, female = 83, median age, 51; range, 20-85 years) were assessed following single-session, large dose (3 x 5 ml) phenol injection therapy. The most frequent complaint was bleeding (100 percent). RESULTS: At four-year follow-up, 53 patients (28.0 percent) were cured, 26 (13.7 percent) were improved, 35 (18.5 percent) remained unchanged, 59 (31.2 percent) deteriorated, and 16 (8.5 percent) required surgical intervention. Among the patients who were not cured, symptoms were minimal in 50 percent. Sclerotherapy was associated with a reduced incidence of bleeding (P < 0.05) but an increase in difficulty in perineal cleaning was observed (P < 0.05). CONCLUSION: Large-dose, single-session sclerotherapy provides only short-term benefits in the majority of patients with symptomatic hemorrhoids. PMID- 8404390 TI - Therapeutic experience of hyperbaric oxygenation in radiation colitis. Report of a case. AB - PURPOSE: The usefulness of the hyperbaric oxygenation in a patient with radiation colitis was evaluated. METHODS: The patient had severe bleeding from the large bowel which resulted from a pelvic radiation therapy for prostatic carcinoma and was histologically diagnosed as radiation colitis. Hyperbaric oxygenation therapy consisting of 100 percent oxygen inhalation at two absolute atmospheric pressures 90 minutes daily for 30 days was performed. RESULTS: The treatment succeeded in stopping gross hemorrhage and reversed endoscopic bowel findings. CONCLUSION: The effect of the hyperbaric oxygenation therapy of radiation colitis is elucidated. PMID- 8404391 TI - Laparoscopic loop ileostomy for temporary fecal diversion. AB - Loop ileostomy is an effective means of temporary fecal diversion. This type of stoma is easy to manage and closure does not require formal laparotomy. We describe a laparoscopic technique of loop ileostomy formation. The procedure can be done with ease and with less discomfort and morbidity when compared with open laparotomy. PMID- 8404392 TI - Single-operator hemorrhoid ligator. AB - A new device for elastic band ligation of hemorrhoids is presented. The three instruments used for this procedure, the anoscope, clamp, and ligator, are combined to allow performance of the procedure by a single operator. PMID- 8404393 TI - Rectodynamics or fecoflowmetry? PMID- 8404394 TI - Use of botulinum toxin in anal fissure. PMID- 8404395 TI - Intestinal motility in irritable bowel syndrome: is IBS a motility disorder? Part 1. Definition of IBS and colonic motility. PMID- 8404396 TI - Intestinal motility in irritable bowel syndrome: is IBS a motility disorder? Part 2. Motility of the small bowel, esophagus, stomach, and gall-bladder. PMID- 8404397 TI - Loperamide abolishes exercise-induced orocecal liquid transit acceleration. AB - Previous work in our laboratory has found that mild physical activity accelerates mouth-to-large intestinal transit of lactulose in a mixed liquid meal. Because loperamide is commonly used as an antidiarrheal agent, we wondered if it would blunt the orocecal transit acceleration provoked by mild exercise. We investigated this equation in 12 healthy persons by comparing orocolonic liquid transit at rest and in mild exercise. Each subject ingested 8 mg loperamide 1 hr prior to study under both resting and exercise conditions. With loperamide treatment, exercise (walking at 5.6 km/hr) failed to hasten increased H2 excretion (mean transit time 72 +/- 12 min at rest, 90 +/- 15 min in exercise; P = NS). This result contrasts sharply with previously reported controls: loperamide completely abolished exercise-induced orocecal transit acceleration ( 23 +/- 5 min in controls; +18 +/- 13 min with loperamide; P < 0.05). Compared with these same controls, resting transit was not significantly slowed by the drug, while transit in exercise was retarded (64 +/- 5 min in controls, 90 +/- 15 min with loperamide; P = 0.06). Loperamide left unchanged the heart rate and oxygen uptake rises associated with exercise. In summary, by showing that loperamide blocks an exercise effect on the upper gut, these results suggest that the drug might prove effective in treating some gut symptoms induced by physical activity. PMID- 8404398 TI - Detection of ploidy in colorectal tumors. A comparison between flow cytometry and cytogenetics. AB - Parallel investigations of ploidy by flow cytometry and cytogenetics were performed in 20 colorectal tumors. Flow cytometry detected an aneuploidy in 13 tumors with DNA indices ranging from 1.13 to 2.21. The other samples exhibited an apparent diploid DNA content. Cytogenetic analyses revealed an abnormal chromosome count in 14 cases and a frequent implication of numerical or structural changes in chromosomes 1, 7, 12, 17, 18, and 20. DNA content evaluated by both techniques was generally concordant with minor discrepancies not exceeding 10%. In six cases, cytogenetics failed to find the cell populations detected by flow cytometry. These results indicate that flow cytometry and cytogenetics are reliable and complementary techniques, particularly in near diploid tumors, where flow cytometry has some difficulties in detecting variations from diploidy below to 8%. PMID- 8404399 TI - Proximal colonic response and gastrointestinal transit after high and low fat meals. AB - The fat component of meals has been thought to make a major contribution to the colonic response to feeding. We have combined gamma scintigraphy and radiotelemetry to noninvasively study the response of the normally inaccessible proximal colon after ingestion of either a high or low fat meal. Separate studies were performed to measure the rate of passage of the same meals through the whole gut. Gastric emptying and small bowel transit of the two meals to the colon was similar, 50% of meal marker reaching the ascending colon 4.8 +/- 0.2 and 4.5 +/- 0.3 hr after the high and low fat meals respectively (N = 8, difference not significant). The low fat meal caused a consistent increase in motility index, which rose from a basal value of 1.0 +/- 0.3 to 2.6 +/- 0.7 mm Hg in the 2 hr after the meal (N = 8, P < 0.01). Response to the high fat meal was less consistent, motility index increasing from 1.6 +/- 0.6 basally to 2.3 +/- 0.7 mm Hg postprandially (N = 8, P = 0.21). Despite these increases in motor activity there was no net caudal propulsion of colonic contents after either meal. The geometric center was comparable, being 3.2 +/- 0.4 and 3.7 +/- 0.4 before the high and low fat meals. This did not change significantly after either meal, being then 3.5 +/- 0.4 and 3.6 +/- 0.4 2 hr after the high and low fat meals, respectively. We conclude that in normal subjects equicaloric high and low fat meals transit the whole gut at a similar rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404400 TI - Constipation after rectopexy for rectal prolapse. Where is the obstruction? AB - The pathophysiology of constipation after rectopexy remains unclear: acquired anorectal dysfunction or preoperative colonic state are, by turns, the supposed culprit. The aim of this prospective study was to characterize the colorectal motility abnormalities encountered after such a surgical procedure. Twelve patients (10 females, 2 males, aged 50.5 +/- 5.2 years) complaining of severe constipation or its worsening after Orr rectopexy (OR) for rectal prolapse were studied. Each underwent detailed interrogation as to their symptoms, left colonic manometry (basal and postprandial motor indexes and their caudad gradients in the sigmoid), anorectal manometry, evacuation proctography, and colonic transit time with radiopaque markers. Results were compared to those obtained in two control groups: 10 healthy volunteers (HV) and 12 patients complaining of a rectal prolapse (RP) observed consecutively during the same period of evaluation (June 90 to December 91). Before surgery, the OR and RP groups were similar with respect to mean age, sex ratio, weekly stool frequency, subjective dyschezia and manual anal supplies, constipation symptoms, and anal incontinence. OR patients differed significantly from the RP group in having a lower weekly stool frequency (2.5 +/- 2.2 vs 5.2 +/- 3.7, P < 0.01) and a higher prevalence of abdominal pain (7 vs 1 patients, P < 0.05). Above the rectopexy, global (135.9 +/- 38 vs 51 +/- 30.5 hr, P < 0.01) and left (61.6 +/- 10 vs 18.2 hr, P < 0.01) colonic transit times were significantly higher in OR patients; moreover, the basal motor index gradient was negative in all but one case (-94.1 +/- 101 vs 177.3 +/- 131, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404401 TI - Effect of sleep quality on symptoms of irritable bowel syndrome. AB - A prospective, one-month diary study was conducted with 23 adult irritable bowel syndrome (IBS) patients in order to determine the relationship between IBS and the quality of sleep. Subjects were screened through history and diagnostic studies. Accepted patients then completed a daily diary of IBS symptoms and sleep quality. At baseline, most subjects (74%) characterized themselves as "poor sleepers." Using pooled time series analysis, the study found a significant correlation between morning IBS symptoms and the quality of the prior night's sleep (P < 0.001), a finding not previously reported in the literature. A less strong but still significant correlation (P < 0.05) was found between end of day IBS symptoms and the quality of sleep during the prior evening. Morning IBS symptoms seem to rise or fall in close association with the prior night's quality of sleep. The study supports the hypothesis that IBS symptoms are related to a disturbance in sleep. PMID- 8404402 TI - The cecum is the site with the highest calcium absorption in rat intestine. AB - In the absence of electrochemical gradients, mucosa-to-serosa Ca transport across the rat cecum is about six times higher than the serosa-to-mucosa flux, resulting in a marked Ca absorption, which is considerably higher when compared with Ca absorption reported for other intestinal segments. The voltage-clamp experiments reveal that 45% of the total mucosa-to-serosa Ca transport measured across the short-circuited tissue is cellular whereas 55% is paracellular. The serosa-to mucosa Ca flux, however, is purely paracellular. Dexamethasone or 1,25(OH)2D3 has no effect on the Ca transport across the cecum. Diphosphonate, known to inhibit 1,25(OH)2D3 synthesis, abolishes cellular mucosa-to-serosa Ca transport but this effect can be restored by simultaneous exogenous 1,25(OH)2D3 application. It is concluded that the cecum is the site with the highest calcium absorption in the rat intestine. The cellular mucosa-to-serosa Ca transport is dependent on 1,25(OH)2D3 but cannot be increased by exogenous 1,25(OH)2D3 application, suggesting that the Ca carrier in this segment already under normal nutritional conditions works at the maximal level. Ca absorption across the proximal colon therefore seems to be of physiological importance in the regulation of intestinal Ca homeostasis. PMID- 8404403 TI - Entamoeba histolytica cyst passers. Clinical profile and spontaneous eradication of infection. AB - The present study was carried out to examine whether Entamoeba histolytica cyst passers suffer from any parasite-related bowel symptoms and to assess the frequency of spontaneous eradication of this infection. The study was carried out in two parts. In part I, stool samples were collected at random from 3536 individuals living in rural communities around Delhi. E. histolytica was isolated by the culture technique in 345 (9.7%) subjects. There was no increase in the prevalence rate of bowel symptoms in the culture-positive compared to the culture negative subjects. One hundred twenty-four (36%) of the culture-positive subjects agreed to take part in a longitudinal study; the subjects were left untreated and clinical assessment and stool examinations were carried out at three-month intervals. One hundred eighteen (95.2%) subjects had eradicated their parasite spontaneously at the end of one year; none developed any features of invasive amebiasis. Part II of the study was carried out on 625 patients attending our Gastroenterology Clinic. Positive cultures of E. histolytica were obtained from 99 (15.2%) patients. Again, there was no increase in the prevalence rate of bowel complaints in the culture-positive compared to the culture-negative subjects. Moreover, histological appearances of the rectal biopsy specimens were not significantly different between the two groups. Twenty-eight (28.2%) patients agreed to the longitudinal study and all eradicated the parasite spontaneously within five months; none developed any evidence of invasive amebiasis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404404 TI - Olsalazine in maintenance of clinical remission in patients with ulcerative colitis. AB - Frequent minor side effects are associated with sulfasalazine. The realization that it is the 5-aminosalicylic acid moiety that is the active component of sulfasalazine and that the side effects are probably due to the sulfapyridine has prompted the search for a similar but safer compound. Olsalazine, consisting of two molecules of 5-ASA without sulfasalazine may avoid the problems due to sulfasalazine. One hundred one patients were entered into a double-blind placebo controlled study of the use of olsalazine (2 g daily) in preventing relapse in patients who had recently recovered from an acute attack of ulcerative colitis. Patients were treated for 12 months. Forty-nine were randomized to olsalazine (39 with limited and 10 with extensive disease) and 52 to placebo (42 with limited and 10 with extensive disease). Life-table analysis showed that the median time to relapse in patients on olsalazine was 342 days, which was significantly longer than the 100 days in the placebo group (P = 0.024). The most important side effect experienced with olsalazine that necessitated withdrawal from the study was "drug-induced diarrhea" in 16% (8/49). There was a similar incidence of minor side effects reported in the two groups, and in no patients were major or dangerous side effects reported. In patients who did not develop diarrhea, olsalazine was well tolerated and successfully prevented rapid relapse in the recently ill patients entered into this study. PMID- 8404405 TI - Better quality of therapy with 5-ASA colonic foam in active ulcerative colitis. A multicenter comparative trial with 5-ASA enema. AB - We evaluated the efficacy, tolerance, and acceptance of a new 5-ASA colonic foam versus 5-ASA liquid enema in the short-term treatment of active ulcerative colitis in a three-week prospective, randomized, investigator-blind study, enrolling 233 patients from 12 outpatient clinics in Italy. In arm 1 of the study, 117 patients with mild attacks received 2 g of 5-ASA as foam or enema at bedtime. In arm 2, 116 patients with moderate attacks were given 4 g of 5-ASA as foam or enema at bedtime. End points were defined as complete relief of symptoms, and endoscopic and histological evidence of remission or improvement. In patients with mild relapse, 34 of 63 (54%) treated with foam were in clinical remission after only 10 days compared with 17 of 51 (31%) treated with enemas (P < 0.05). However, there was no statistically significant difference between foam (83%) and enema (74%) after three weeks. In patients with moderate relapse, a higher proportion of patients achieved complete clinical remission in the foam group (63%) compared with enema group (52%) after three weeks (difference 11%, 95% CI 7 to 29). No significant differences were observed in endoscopic and histological evaluation of colonic mucosa between treatment groups in either arm. 5-ASA foam was well tolerated. No unexpected adverse events were reported. Patient evaluation of therapy showed that foam was much better accepted than enema because foam was more comfortable, more practical, easier to retain, and interfered less with daily living.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404406 TI - Low-dose oral methotrexate in refractory inflammatory bowel disease. AB - The purpose of this study was to evaluate the efficacy and safety of low-dose weekly, oral methotrexate in patients with steroid-dependent or steroid refractory inflammatory bowel disease (IBD). Oral methotrexate was given weekly at 15 mg/week. The primary criterion of response was based on steroid withdrawal. Of the 10 patients with Crohn's disease, daily prednisone dosage dropped from a mean of 37 +/- 9.6 mg to 8.3 +/- 2.1 mg/day (P < 0.02); two had a complete withdrawal and four a partial response (< 7.5 mg/day). In the eight patients with ulcerative colitis, daily prednisone dose dropped from a mean of 26.3 +/- 3.2 mg/day to 12.7 +/- 2.0 mg/day (P < 0.001); three had a partial response. Adverse effects due to methotrexate were mild in both groups. We conclude that oral methotrexate may be useful and reasonably safe as a steroid-sparing agent in patients with refractory IBD. PMID- 8404407 TI - Catecholamine concentrations in biopsied gastroduodenal tissue specimens of patients with duodenal ulcer. AB - We measured dopamine and norepinephrine concentrations in the biopsied gastroduodenal mucosa obtained from 12 ulcer-free dyspeptic patients, nine patients with active duodenal ulcer, and eight patients with inactive (or healed) duodenal ulcer using a high-performance liquid chromatography with electrochemical detection method. Biopsy specimens were taken from endoscopically normal-appearing mucosa in the gastric body and antrum as well as in the duodenal bulb. Additional specimens were obtained from the outer edge of the ulcer margin in patients with active duodenal ulcer. The mean (+/- SD) mucosal dopamine concentrations in the gastric body and duodenum (7.6 +/- 2.8 and 6.8 +/- 2.6 pg/mg tissue) obtained from patients with inactive duodenal ulcer were significantly (P < 0.05) lower than those from dyspeptic patients (13.6 +/- 6.9 and 10.9 +/- 3.5 pg/mg tissue, respectively). In contrast, no significant differences were observed in the mean norepinephrine concentrations in these gastroduodenal tissues among the three study groups. However, the mean mucosal norepinephrine concentration in the outer edge of duodenal ulcer (86.2 +/- 125.6 pg/mg tissue) was significantly (P < 0.05 and 0.01) reduced as compared with that in the ulcer-free area of duodenum obtained from patients with inactive duodenal ulcer (257.1 +/- 188.2 pg/mg tissue) and from dyspeptic patients (276.8 +/- 138.3 pg/mg tissue). The results suggest that an alteration in the catecholaminergic system may be associated with one of the pathogenic factors of duodenal ulcer. PMID- 8404408 TI - Vagal effects on acid and pepsin secretion and serum gastrin in duodenal ulcer and controls. AB - To study whether the vagus sustains basal secretion and stimulates acid and pepsin differently in duodenal ulcer (DU) and non-DU, we tested 144 patients with DU and 92 nonulcer controls, using 1-hr basal secretion followed by 15 min of modified sham feeding (MSF) and after 1 hr followed by a reference maximum elicited by 6 micrograms/kg pentagastrin given subcutaneously and observed for another 1 hr. Of all subjects, 97.5% responded to MSF by raising basal acid output (BAO) at least 15%. MSF added amounts of acid equal to 26-30% of peak acid output and 30-43% of peak pepsin output, regardless of diagnosis or level of basal secretion (including hypersecretors). Speed and duration of responses were similar in DU and controls. MSF did not substantially alter serum gastrin. Males secreted more acid and pepsin than females under all conditions, differences that persisted in DU but not in controls when outputs were corrected for body weight. Male DU but not female DU patients secreted more than corresponding controls. Sham feeding is an effective stimulus with similar characteristics in controls and DU patients. There was no evidence for saturation of vagal pathways in basal hypersecretors. MSF stimulation does not appear to involve gastrin. Hypersecretion in DU derives largely from responses in male DU. PMID- 8404409 TI - Accumulation of aliphatic amines in gastric juice of acute renal failure patients. Possible cause of hypergastrinemia associated with uremia. AB - In this study we analyzed by gas chromatographic headspace analysis the composition and concentration of gastrin-stimulatory volatile aliphatic amines in the gastric juice of healthy subjects and acute renal failure patients. We demonstrated that although these aliphatic amines are present in the gastric juice of normal subjects in trace amounts, they accumulate in the gastric juice of uremic subjects. This 30-40-fold elevation in gastric juice amine concentration agreed favorably with the 40-50-fold augmentation in serum gastrin levels in acute renal failure, with a significant association (r = 0.87) existing between these two parameters. It was also determined that a 2-hr hemodialysis procedure resulted in a modest nonparallel decline in both gastric amine and serum gastrin levels. These results support the hypothesis that the accumulation of volatile aliphatic amines in the gastric juice of uremic individuals may induce an activation of the antral G cells, resulting in hypergastrinemia. PMID- 8404410 TI - Effect of cholestyramine and cholecystokinin receptor antagonist CR1505 (loxiglumide) on lower esophageal sphincter pressure in man. AB - Cholecystokinin (CCK) decreases lower esophageal sphincter pressure (LESP) in man. Cholestyramine, a nonabsorbable bile salt binding resin, stimulates endogenous CCK secretion. We have investigated the effect of oral ingestion of 16 g cholestyramine without and with infusion of the CCK receptor antagonist CR1505 (loxiglumide, 15 mg/kg/90 min) on LESP in seven healthy subjects. LESP was recorded for 90 min, in 10-min intervals, with the pull-through technique using a four-lumen water-perfused catheter. Oral ingestion of cholestyramine resulted in a significant (P < 0.05) decrease in LESP, starting from 10 min and continuing during the entire experiment (basal LESP: 11.8 +/- 2 mm Hg, minimal value reached during cholestyramine: 7.3 +/- 1 mm Hg; P < 0.05). Pretreatment with loxiglumide completely antagonized the effect of cholestyramine on LESP. Infusion of loxiglumide without cholestyramine did not affect basal LESP. It is concluded that: (1) cholestyramine significantly reduces LESP; (2) this reduction in LESP does not occur after pretreatment with loxiglumide, indicating that cholestyramine influences LESP through CCK release; and (3) basal LESP is not regulated by CCK. PMID- 8404411 TI - Complications during pneumatic dilation for achalasia or diffuse esophageal spasm. Analysis of risk factors, early clinical characteristics, and outcome. AB - A retrospective cohort study was performed to assess risk factors, early clinical characteristics, and outcome of complications in patients undergoing pneumatic dilation. Of 178 patients with achalasia or diffuse esophageal spasm who underwent 236 dilations with a Browne-McHardy dilator, 16 patients experienced a complication (9.0%). Nine major complications developed: perforations (4), hematemesis (2), fever (2), and angina (1). A prior pneumatic dilation and use of inflation pressure > or = 11 PSI were independent risk factors by multivariate analysis for developing a complication. An esophagram immediately following the dilation identified three of the four perforations. Three postdilation findings were identified as indicators of patients with an increased risk of having developed a perforation: blood on the dilator, tachycardia, and prolonged chest pain lasting > 4 hr after dilation. In all patients incurring a major complication, one of the three indicators, or the complication itself was recognized within 5 hr of dilation. All patients with complications, including the four with perforation who received prompt surgical repair and esophagomyotomy, recovered uneventfully. The symptomatic relief of dysphagia in patients with perforation undergoing emergent surgical repair and esophagomyotomy was similar to patients undergoing elective esophagomyotomy. CONCLUSIONS: (1) Pneumatic dilation is a safe treatment of achalasia, with a 1.7% risk of perforation. (2) The risk of developing a complication is increased by having had a previous pneumatic dilation or by use of inflation pressures > or = 11 psi. (3) All patients with a major complication were identified within 5 hr after dilation. (4) Complications following pneumatic dilation, if recognized and treated promptly, were not associated with adverse, long-term sequelae. PMID- 8404412 TI - Achalasia. Short-term clinical monitoring after pneumatic dilation. AB - Although concern about perforation has led physicians to perform pneumatic dilation for achalasia with routine contrast radiography immediately afterwards and with hospitalization, the need for these precautions has not been demonstrated. In contrast, we have routinely performed pneumatic dilations without contrast studies or hospitalization, and we hereby present our experience. During a recent six-year period, 110 pneumatic dilations were performed, and 71 of the last 73 were performed as outpatients with about 5-8 hr of clinical monitoring. Detailed review of 100 records showed that only 15 patients underwent contrast studies because of pain or fever. Perforation occurred in seven of the 15 patients, all of whom underwent surgery successfully. Short-term follow-up in patients who did not sustain perforation showed good or excellent results in 82%. Thus, it has been our experience that pneumatic dilation could be safely performed in achalasia without routine use of contrast studies or hospitalization. PMID- 8404413 TI - Role of upper esophageal reflex and belch reflex dysfunctions in noncardiac chest pain. AB - Fifty-four patients examined for noncardiac chest pain (NCCP), showing no esophageal motor disorder or gastroesophageal reflux disease compatible with NCCP, were subjected to an intraesophageal balloon distension test and a study of the belching reflex provoked by intraesophageal air injection. Thirty-three control subjects were also studied, allowing us to define high-threshold belchers (group I) as those who belched during two of three 40-ml distensions and low threshold belchers (group II) as those who did not. The balloon distension test induced NCCP in 64% of the patients in group I, and in 14% of the patients in group II (P < 0.01). High-threshold belching was a factor favoring the positivity of the balloon distension test. This result supports the hypothesis that esophageal distension by air due to a belching disorder may be the mechanism responsible for NCCP in some patients with an abnormal sensitivity to balloon distension. PMID- 8404414 TI - pH profile of esophagus in patients with inlet patch of heterotopic gastric mucosa after tetragastrin stimulation. An endoscopic approach. AB - In a series of 1771 endoscopic examinations from 1988 to 1990, we observed 62 cases (3.3%) of inlet patch of heterotopic gastric mucosa (IPHGM) in the upper esophagus. Ten of the IPHGM patients complained of throat discomfort or globus sensation, and these symptoms were relieved by histamine H2-antagonists, suggesting that these symptoms could be caused by acid secretion from IPHGM. Acidity under tetragastrin stimulation was measured in the esophagus and compared with the pool of gastric juice and saliva under direct vision by a newly devised endoscopic method. Congo red staining was also carried out after pH measurement. Significant pH reduction at IPHGM was observed in three cases, and black coloration with Congo red staining in the IPHGM was observed in four cases. These findings suggest that IPHGM is a mucosal change with latent acid secretion. PMID- 8404415 TI - Epidemiology of achalasia in central Israel. Rarity of esophageal cancer. AB - The epidemiology of achalasia was studied in a predominantly urban, Jewish population of approximately 1.3 million, in central Israel, during the years 1973 1983. One hundred sixty-two proven cases were collected, representing all known patients with achalasia in the study area. There were no gender differences. The majority of cases were diagnosed within two years of onset of symptoms, although the median delay in diagnosis was 4.4 +/- 5.3 years. The disease was rare in the first two decades of life. The prevalence (in 1983) in the first two decades was 0.7/10(5) rising to 36.2/10(5) above age 70. The mean annual incidence in the years 1973-1978 was 0.8/10(5). It rose slightly to a mean annual incidence of 1.1/10(5) in the years 1979-1983. The prevalence of the disease in 1973 and 1983 was 7.9/10(5) and 12.6/10(5), respectively. The age-adjusted prevalence in 1973 was higher in Asian and African born Jews as compared to those born in Europe, America, or Israel. This difference disappeared by the year 1983. No case of cancer of the esophagus was found among our patients. This may be due to the nonselected, regional nature of our series or to the effects of earlier therapy of achalasia in recent decades. PMID- 8404416 TI - Age- and gender-related differences in 24-hour esophageal pH monitoring of normal subjects. AB - Twenty-four-hour esophageal pH monitoring is currently the most sensitive test for diagnosing gastroesophageal reflux. Little is known, however, about the effect of aging and gender on esophageal acid exposure in asymptomatic individuals. Thirty asymptomatic volunteers underwent 24-hr esophageal pH monitoring. Fifteen were < 65 years (eight female, seven male) and 15 were > or = 65 years (seven female, eight male). In this asymptomatic group no significant difference was seen by age, while males were found to have significantly more esophageal acid exposure than females. The need for sex-specific normal 24-hr pH monitoring values is suggested. Thirty percent of these asymptomatic subjects were abnormal by conventional 24-pH criteria. The clinical importance of these "silent refluxers" is unknown. PMID- 8404417 TI - Technical aspects of nonoperative dilation of a complex colon anastomotic stricture. PMID- 8404418 TI - Intussusception and leiomyosarcoma of the gastrointestinal tract in a pediatric patient. Case report and review of the literature. AB - Intestinal leiomyosarcoma is a rare tumor in infants and children; only 46 cases have been reported in the English literature. Presenting signs and symptoms include abdominal pain and gastrointestinal obstruction and bleeding. We describe a neonate with the unique presentation of ileocecal intussusception accompanying an ileal leiomyosarcoma. In contrast to adult patients, where intussusception is associated with smooth muscle tumors in 30% of cases, leiomyosarcoma and subsequent intussusception is rare in infancy and childhood. The overall prognosis for long-term survival is similar for both pediatric and adult patients with leiomyosarcoma of the intestinal tract, with a five-year survival in reported cases of 53% and 40%, respectively. PMID- 8404419 TI - Ileal involvement in toxic epidermal necrolysis (Lyell syndrome). AB - Intestinal involvement in toxic epidermal necrolysis (TEN) has been identified only rarely. This report describes a case of TEN with ileal manifestations characterized by a profuse diarrhea with malabsorption, protein-losing enteropathy, and radiologically by multiple stenosis. After healing of the cutaneous lesions, total parenteral nutrition was initiated, resulting in a decreased diarrhea. However, after one month of total parenteral nutrition, malabsorption and protein-losing enteropathy continued and the radiological lesions were still present with an aspect consistent with a sclerotic process. A surgical resection of the pathological ileal segment was performed with end-to end anastomosis. Pathological examination of the resected segment showed a necrosis of the ileal mucosa with a pattern similar to that of the epidermal necrosis. No sclerosis was observed. It seems that a prolonged total parenteral nutrition could have induced a complete healing of intestinal lesions. This case report is the first clinical, radiological, and histological study of an ileal involvement in TEN. PMID- 8404420 TI - Clostridium difficile enteritis. A cause of intramural gas. PMID- 8404421 TI - Leiomyomatous colonic polyp. AB - Leiomyomatous colonic polyps are rare. We report a patient with an asymptomatic 15-mm submucosal polyp in the sigmoid colon. The tumor was successfully removed by endoscopy. Histopathological examination revealed a leiomyoma originating in the muscularis mucosa. PMID- 8404422 TI - Intragastric pH and upper gastrointestinal bleeding in the ICU. PMID- 8404423 TI - Medical treatment of phytobezoar. PMID- 8404424 TI - Effectiveness, safety, and acceptability of a copper intrauterine device (CU Safe 300) in type I diabetic women. AB - OBJECTIVE: To evaluate the long-term effectiveness and safety of a copper intrauterine device fulfilling modern standards in type I diabetic women compared with nondiabetic women. RESEARCH DESIGN AND METHODS: Type I diabetic women (n = 59, age 27 +/- 5 yr, duration of diabetes 12 +/- 8 yr, HbA1c 7.0 +/- 1.2%, 78% nulliparous women) were prospectively evaluated at 3, 6, 12, 24, and 36 mo by a gynecological exam and a standardized questionnaire after insertion of the intrauterine device (CU Safe 300, 300 mm2 of copper). A group of nondiabetic women (n = 1150) of comparable age and parity evaluated according to the same study protocol served as a control group. RESULTS: In the diabetic women (1754 cumulative months of use), events leading to termination of the intrauterine device during the 1st yr (691 women-mo) were one accidental pregnancy, one expulsion, one removal for pain, two removals for bleeding, and one removal for planned pregnancy. Events during the 2nd (593 women-mo) and 3rd yr (470 women-mo) were zero and one accidental pregnancy., one and two removals for bleeding, one and one removal for pain, one and one removal for other medical reasons, and two and two removals for planned pregnancy, respectively. No case of pelvic inflammatory disease was diagnosed in the diabetic group, and one case was diagnosed in the nondiabetic group (28,369 mo of cumulative use). Events leading to termination of the intrauterine device per women observed per year and continuation with the intrauterine device after each year of use were comparable in the diabetic and nondiabetic groups for the 1st, 2nd, and 3rd yr. CONCLUSIONS: These results, although preliminary because < 100 diabetic women were studied, indicate that the intrauterine device CU Safe 300 is as effective, safe, and well tolerated in diabetic as in nondiabetic women. Specific objections to the use of intrauterine devices in type I diabetic women do not seem to be justified for modern, copper-mediated models. PMID- 8404425 TI - Diabetes in pregnancy in Zuni Indian women. Prevalence and subsequent development of clinical diabetes after gestational diabetes. AB - OBJECTIVE: To determine the prevalence of gestational diabetes mellitus in Zuni Indian women and the subsequent rate of diabetes among Zuni women with GDM. RESEARCH DESIGN AND METHODS: A retrospective analysis of 809 deliveries over a 4 yr period among Zuni Indian women was conducted to determine the prevalence of GDM and diabetes antedating pregnancy. A prospective case-control study of 47 full-blooded Zuni Indian women with GDM and 47 control subjects was performed to determine the progression to clinical diabetes in women with a first-time diagnosis of GDM. Cases with GDM delivered during a defined 8-yr period. The control group of Zuni women delivered during the same time period but had plasma glucose values < 7.8 mM on the 1-h glucose screening test. Cases with GDM and control subjects were matched for age, body mass index, gravidity, and length of follow-up. All women were re-evaluated for diabetes up to 9 yr after the index pregnancy. RESULTS: Between 1987-1990, 116 cases of GDM and 8 cases of pre existing diabetes were identified, giving a prevalence of maternal diabetes in pregnancy of 15.3%. At the time of follow-up, 14 of 47 (30%) women with GDM had developed diabetes after a mean of 4.8 yr compared with only 3 of 47 (6%) from the control group with an average of 5.5 yr follow-up. CONCLUSIONS: GDM is prevalent among Zuni Indians and is associated with an increased risk of diabetes. Glucose tolerance after GDM may deteriorate at a greater rate in Native Americans than in other populations. PMID- 8404426 TI - A cold spot of IDDM incidence in Europe. Macedonia. AB - OBJECTIVE: To determine, for the first time, the incidence of IDDM among children 0-15 yr of age in Macedonia, the south republic of former Yugoslavia, and to compare these rates with those from neighboring countries. RESEARCH DESIGN AND METHODS: Children < 15 yr of age, with IDDM diagnosed between January 1985 and December 1991, were registered using the criteria established for the WHO Multinational Project for Childhood Diabetes. The primary source of case ascertainment consisted of clinical records from the University Pediatric Clinic in Skopje, the capital of Macedonia. The secondary source consisted of records for insulin supplies from the pharmacy in Skopje and from 15 local hospitals. RESULTS: During 1985-1991, 112 children < 15 yr of age were diagnosed with IDDM in Macedonia. The completeness of case ascertainment was 96.6%. The age-adjusted IDDM incidence rate was very low, only 2.45/100,000 per yr. No significant sex difference in incidence was observed. The highest incidence rate occurred in the 10- to 15-yr age-group. No temporal trends were observed. CONCLUSIONS: The incidence of IDDM in Macedonia is the lowest in Europe and among the lowest reported for Caucasian populations. However, the descriptive epidemiology is similar to that for high incidence populations. Further etiological studies are required to evaluate potential reasons for the very low incidence of IDDM in Macedonia. PMID- 8404427 TI - Alterations in glucose metabolism in the elderly patient with diabetes. AB - OBJECTIVE: To determine the alterations in glucose metabolism in elderly patients with NIDDM. RESEARCH DESIGN AND METHODS: We studied 9 healthy elderly control subjects (73 +/- 1 yr of age; body mass index 25.7 +/- 0.4 kg/m2) and 9 untreated elderly NIDDM patients (72 +/- 2 yr of age; BMI 25.9 +/- 0.5 kg/m2). Each subject underwent a 3-h oral glucose tolerance test (40 g/m2); a 2-h hyperglycemic glucose clamp study (glucose 5.4 mM above basal); and a 4-h euglycemic insulin clamp (40 mM.m2.min-1). Tritiated glucose methodology was used to measure glucose production and disposal rates during the euglycemic clamp. RESULTS: Patients with NIDDM had a higher fasting glucose (9.3 +/- 0.3 vs. 5.1 +/- 0.1 mM in control subjects vs. NIDDM patients, respectively, P < 0.001) and a greater area under the curve for glucose during the OGTT (16.0 +/- 0.6 vs. 6.7 +/- 0.3 mM in control subjects vs. NIDDM patients, respectively, P < 0.01) than the healthy control subjects. During the hyperglycemic clamp, patients with NIDDM had an absent first phase insulin response (112 +/- 6 vs. 250 +/- 31 pM in control subjects vs. NIDDM patients, respectively, P < 0.01), and a blunted second-phase insulin response (159 +/- 11 vs. 337 +/- 46 pM in control subjects vs. NIDDM patients, respectively, P < 0.01). Before the euglycemic clamp, fasting insulin (99 +/- 5 vs. 111 +/- 10 pM in control subjects vs. NIDDM patients, respectively) and hepatic glucose production (11.8 +/- 0.7 vs. 11.5 +/- 0.5 mumol.kg-1-min-1 in control subjects vs. NIDDM patients, respectively) were similar. Steady-state (180-240 min) glucose disposal rates during the euglycemic clamp were slightly, but not significantly, higher in the normal control subjects (36.5 +/- 1.1 vs. 33.1 +/- 1.9 mumol.kg-1-min-1 in control subjects vs. NIDDM patients, respectively, NS). CONCLUSIONS: We conclude that NIDDM in nonobese elderly subjects is characterized by a marked impairment in insulin release. This may be attributable to the toxic effects of chronic hyperglycemia on the beta-cell. When compared with age-matched control subjects, the NIDDM patients showed no increase in fasting insulin or hepatic glucose production, and insulin resistance was mild. PMID- 8404428 TI - Is the corrected QT interval a reliable indicator of the severity of diabetic autonomic neuropathy? AB - OBJECTIVE: We investigated whether the corrected QT interval correlated with two other tests for diagnosing autonomic dysfunction in 60 type I diabetic patients with proven peripheral neuropathy. The mean age +/- SD was 48.3 +/- 11.2 yr, the mean duration of diabetes was 24.9 +/- 11.4 yr, and the mean HbA1 was 9.3 +/- 2.4%. RESEARCH DESIGN AND METHODS: All patients underwent three autonomic function tests: 1) the standard five cardiovascular Ewing tests, each scored 0 (normal), 0.5 (borderline), or 1.0 (abnormal). We used the sum of the abnormal findings for the analysis, the cardiovascular autonomic score; 2) measurement of the corrected QT interval taken from a routine electrocardiogram recording; and 3) static and dynamic pupillometry: measurement of dark adapted pupil diameter as percentage of total iris diameter and of pupil constriction latency using an infrared light reflex technique. RESULTS: No significant correlation was found between age, duration of diabetes, or HbA1 and any of the autonomic function tests, except for one between age and cardiovascular autonomic score (r = 0.3202, P = 0.0126). Corrected QT interval did not correlate with cardiovascular autonomic score, pupil diameter, or constriction latency. A significant inverse correlation was found between cardiovascular autonomic score and pupil diameter (r = -0.4861, P < 0.001) and constriction latency (r = 0.3783, P < 0.001). Pupil diameter and constriction latency correlated well (r = -0.4276, P < 0.001). CONCLUSIONS: The corrected QT interval did not correlate with cardiovascular autonomic tests nor pupillometry results. The corrected QT interval therefore should not be used for the diagnosis of the severity of diabetic autonomic neuropathy. PMID- 8404429 TI - Administration of sulfonylureas can increase glucose-induced insulin secretion for decades in patients with maturity-onset diabetes of the young. AB - OBJECTIVE: To ascertain whether the effect of sulfonylureas on glucose-mediated insulin release persists for years to decades in patients with maturity-onset diabetes of the young. RESEARCH DESIGN AND METHODS: The effect of sulfonylurea treatment on glucose-induced insulin secretion was ascertained prospectively for up to 33 yr in 12 diabetic patients of the maturity-onset diabetes of the young RW pedigree, who are genetically homogeneous because they share DNA markers on chromosome 20q. In 7 of these patients, paired glucose tolerance tests, given while the patients were on and off sulfonylureas, were performed after 7-31 yr. RESULTS: Glucose-induced insulin secretion showed an average increase of 68% in diabetic patients who remained responsive to chlorpropamide after having been on and off the drug for decades. In most patients, however, glucose-induced insulin secretion declines over time (1-4%/yr). Some patients become unresponsive to sulfonylureas after 3-25 yr and then have very small or no increases in glucose induced insulin secretion and require treatment with insulin to normalize fasting hyperglycemia. CONCLUSIONS: Increase in glucose-induced insulin secretion remains the most important mechanism of the action of sulfonylureas during long-term administration. PMID- 8404430 TI - Differences in the prevalence of diabetes and impaired glucose tolerance according to maternal or paternal history of diabetes. AB - OBJECTIVE: To determine whether diabetes risk is influenced by which parent (a parental history of diabetes is a well-documented risk factor for NIDDM) is reported to have diabetes. RESEARCH DESIGN AND METHODS: We compared the prevalence of NIDDM and IGT for 4914 subjects according to their parental history of diabetes (mother only, father only, both parents, neither parent). Subjects were drawn from the San Antonio Heart Study, a population-based survey of diabetes and cardiovascular risk factors conducted in Mexican American and non Hispanic white individuals between 1979-1988. RESULTS: Men with a parental history of diabetes had a higher prevalence of both NIDDM and impaired glucose tolerance than men reporting no parental history of diabetes. Prevalence was equally high regardless of which parent, or whether both parents, had diabetes. In contrast, in women, only a maternal history of diabetes was associated with a higher prevalence of NIDDM and impaired glucose tolerance. Virtually no difference in NIDDM prevalence was found between women with a paternal-only history of diabetes and women with no parental history of diabetes. CONCLUSIONS: Results differed markedly between men and women. The reason for this sex difference is unclear. It may represent a measurement bias, a sex-specific environmental effect, or a genetic effect that is expressed or transmitted differently between the sexes. PMID- 8404431 TI - Evaluation of a structured treatment and teaching program for non-insulin-treated type II diabetic outpatients in Germany after the nationwide introduction of reimbursement policy for physicians. AB - OBJECTIVE: To evaluate the practicability and efficacy of a structured treatment and teaching program for non-insulin-treated type II diabetic patients in routine primary health care. RESEARCH DESIGN AND METHODS: All physicians (n = 139) and their office staffs in Hamburg, Germany, who had participated in a special training course from 1 April 1991 to 31 December 1991 were contacted for a standardized interview. A random sample of 17 of these offices was selected for office visits during which the documented data of all patients who had received the standardized treatment and teaching in the same period were collected and evaluated. RESULTS: The program was well received by the physicians, and the data collected on 179 patients (5.1 mo median after the intervention) demonstrated the efficacy of the program at the treatment level: reduction of body weight (mean 2.8 kg, P < 0.0001) and HbA1c levels (from 8.11 +/- 1.68 to 7.47 +/- 1.64%, P < 0.0001) was substantial. The individual prescribed volume of oral antidiabetic agents was approximately 50% lower after patient attendance of the program (significant decrease from 1.41 +/- 1.42 to 0.76 +/- 1.11 tablets/patient/day, P < 0.0001). The proportion of patients treated with oral antidiabetic drugs decreased from 63 to 42% (P < 0.0001). CONCLUSIONS: Subsequent to the introduction of nationwide remuneration of outpatient education for type II diabetic patients by office-based physicians, a relevant improvement was observed in the quality of care, comparable with the effects of the program in a previous prospective controlled trial. PMID- 8404432 TI - Clinical gallbladder disease in NIDDM subjects. Relationship to duration of diabetes and severity of glycemia. AB - OBJECTIVE: To examine the relationship between the prevalence of gallbladder disease and severity of glycemia among diabetic individuals and to provide insight into whether the diabetes-gallstone association is a causal one, because NIDDM patients have an increased prevalence of clinical gallbladder disease. RESEARCH DESIGN AND METHODS: We examined 462 diabetic individuals identified during the San Antonio Heart Study, a population-based survey of diabetes and cardiovascular disease in Mexican Americans and non-Hispanic whites. Diabetes was diagnosed according to National Diabetes Data Group criteria. RESULTS: The prevalence of self-reported gallbladder disease was 34.2% in diabetic women and 7.2% in diabetic men. Although duration of diabetes was positively related to the prevalence of gallbladder disease (P < 0.01), type of therapy was not associated, and fasting glucose concentration was inversely associated with gallbladder disease. CONCLUSIONS: Factors other than hyperglycemia may account for the increased prevalence of gallbladder disease in diabetic subjects. PMID- 8404433 TI - Improvement of lipoprotein lipid composition in type II diabetic patients with concomitant hyperlipoproteinemia by acipimox treatment. Results of a multicenter trial. AB - OBJECTIVE: To study the tolerability and efficacy of acipimox on hyperlipidemia and diabetes compensation in patients with NIDDM under conditions of a routine clinical practice. RESEARCH DESIGN AND METHODS: We recruited 121 patients (60 men and 61 women) from 10 participating clinical centers. They were randomly divided into two groups and treated for 3 mo either with acipimox (250 mg three times a day) or placebo, using an open study design. RESULTS: Acipimox treatment led to a significant drop in fasting serum total triglyceride levels (by 28%) after 1 mo of drug administration. This decrease prevailed up to the end of the 3-mo study. Serum total cholesterol levels declined by 14%, and high-density lipoprotein tended to rise in acipimox-treated patients. These changes in lipid metabolism were not accompanied by any adverse effects of acipimox on glucose metabolism as judged by HbA1c measurements and the oral glucose tolerance test. Eight patients (out of 82 treated with acipimox) reported moderate adverse events of transient character, such as skin reactions and gastric disturbances. CONCLUSIONS: Acipimox seems to be a useful agent for treatment of diabetic dyslipidemia and does not deteriorate glycemic control. PMID- 8404434 TI - Long-term glycemic control and neurological function in IDDM patients. AB - OBJECTIVE: To examine the relationship between sensory modalities of neurological function and antecedent glycemic control in IDDM patients. RESEARCH DESIGN AND METHODS: Examinations were conducted on 220 IDDM patients (age at onset < 25 yr, duration < 18 yr) for the presence or absence of the right or left ankle reflex and determination of vibration perception threshold at each medial malleolus and great toe using biothesiometry. These parameters were related to the concurrent HbA1 and to a mean of serial measurements (mean HbA1) over the previous 6 yr. RESULTS: Ankle reflexes were absent in 39 (right ankle) and 41 (left ankle) patients, respectively. Mean (right + left) ankle and toe VPTs were 8.7 +/- 3.6 and 6.3 +/- 4.2 (mean +/- SD) (arbitrary units), respectively. Both the mean and concurrent HbA1 were significantly different in patients with absent ankle reflexes (11.6 +/- 1.9 and 12.2 +/- 2.8%, respectively) compared with present ankle reflexes (10.3 +/- 1.7, 10.3 +/- 2.1%) (P < 0.0001). Similarly, a present ankle reflex was related to mean HbA1 arbitrarily divided into groups < 10, 10 12, > 12% (P = 0.0009). In contrast, mean ankle VPT (8.0 +/- 2.2, 8.8 +/- 3.1, and 10.3 +/- 6.2) and toe VPT (5.5 +/- 2.2, 6.1 +/- 2.9, and 8.5 +/- 8.2) did not increase significantly with poor glycemic control (P > 0.05). Age, right ankle reflex, retinopathy, 24-h urinary albumin excretion rate, and erect systolic blood pressure were the only independent variables predicting the toe VPT using linear regression analysis. CONCLUSIONS: These findings support a role for glycemic control in neurological dysfunction in IDDM patients, but also suggest that other unknown factors may be involved. PMID- 8404435 TI - Factitious diabetes mellitus and spontaneous hypoglycemia. Consequences of unrecognized Munchausen syndrome by proxy. AB - OBJECTIVE: To increase health-care professionals' awareness and knowledge of factitious illness by proxy, or Munchausen syndrome by proxy, in relatives of diabetic patients. RESEARCH DESIGN AND METHODS: A case report is described of a 14-yr-old male who has a 6-yr history of diet-controlled diabetes mellitus, and a 6-mo history of reported spontaneous hypoglycemia. RESULTS: Neither diabetes nor spontaneous hypoglycemia was present in this child on objective testing. The child was subjected to inappropriate use of a strict diabetic diet and daily glucometer measurements for at least 8 yr. The father had convinced his son and health-care professionals of these diagnoses, in spite of evidence of the contrary. CONCLUSIONS: When confronted with history and clinical findings that contradict laboratory findings, health-care professionals should have a high index of suspicion for factitious illness and should pursue it aggressively with the help of legal services. PMID- 8404436 TI - Ultrasonographic abnormalities of the pancreas in IDDM and NIDDM patients. AB - OBJECTIVE: To evaluate the relationship between the type and duration of diabetes and pancreas size by ultrasonography. RESEARCH DESIGN AND METHODS: Pancreas images of 40 IDDM and 36 NIDDM patients with 0.3-34 yr of disease were compared with those of 60 normal healthy control subjects. RESULTS: The diameters +/- SD of the head, body, and tail of the pancreas in IDDM patients (1.9 +/- 0.3; 0.9 +/ 0.2; and 1.4 +/- 0.2 cm, respectively) were smaller than in NIDDM patients (2.7 +/- 0.4; 1.2 +/- 0.3; and 1.8 +/- 0.4 cm, respectively) and control group subjects (2.4 +/- 0.4; 1.1 +/- 0.3; and 1.8 +/- 0.4 cm, respectively). The pancreatic shrinkage in IDDM patients was clearly evident after 10 yr of the disease. NIDDM patients and control subjects had similar pancreatic dimensions, except for a greater body thickness in NIDDM patients with > 10 yr of disease (1.2 +/- 0.4 vs. 1.1 +/- 0.3 cm). These results were not related to differences in age, sex, and body size. Pancreas image was hypoechogenic in 72.5% of IDDM patients and hyperechogenic in 83.3% of NIDDM patients. CONCLUSIONS: Smaller pancreases in IDDM patients in comparison with NIDDM patients and control subjects were clearly demonstrated only after 10 yr of disease. Patients with NIDDM were not affected by pancreatic dimensions, except for a greater body thickness after 10 yr of disease. Pancreatic echogenicity increased with age. PMID- 8404437 TI - Insulinoma in a patient with NIDDM. AB - OBJECTIVE: To confirm insulinoma as the cause of hypoglycemia in a patient with NIDDM and determine the frequency of the co-occurrence of these two conditions. RESEARCH DESIGN AND METHODS: The patient underwent an in-hospital prolonged fast (< or = 72 h), according to standard protocol, and an ultrasound examination of the pancreas. All cases of histologically confirmed insulinoma at this institution over the period of 1927-1992 were reviewed to determine the prevalence of pre-existent diabetes mellitus. RESULTS: After 10 h of fasting, plasma glucose was low (1.89 mM); plasma insulin (258 pM) and C-peptide (1.39 nM) were elevated in the absence of sulfonylurea in the plasma. An insulinoma detected by ultrasonography was removed surgically with subsequent reoccurrence of insulin-requiring diabetes. Among 313 cases of insulinoma confirmed at this institution, this patient is the only one with pre-existent diabetes mellitus. CONCLUSIONS: Insulinoma occurs extraordinarily rarely in patients with pre existing NIDDM. PMID- 8404438 TI - Metabolic effects of dietary sucrose in type II diabetic subjects. AB - OBJECTIVE: To assess in diabetic subjects the effects of dietary sucrose on glycemia and lipemia. RESEARCH DESIGN AND METHODS: Twelve type II diabetic subjects consumed, in random order, two isocaloric, 55% carbohydrate study diets for 28 days. In one diet, 19% of energy was derived from sucrose. In the other diet, < 3% of energy was derived from sucrose, and carbohydrate energy came primarily from starch. Both study diets were composed of common foods. All meals were prepared in a metabolic kitchen where foods were weighed during meal preparation. RESULTS: No significant differences were noted between the study diets at any time point in mean plasma glucose. At day 28, mean plasma glucose values for the sucrose diet were 9.6 +/- 0.5 mM and for the starch diet were 9.4 +/- 0.6 mM (P = 0.63). Also, no significant differences were observed between the study diets in urine glucose, fasting serum total, HDL, or LDL cholesterol; fasting serum TG; or peak postprandial serum TG. CONCLUSIONS: A high sucrose diet did not adversely affect glycemia or lipemia in type II diabetic subjects. PMID- 8404439 TI - Quality control in patient self-monitoring of blood glucose. PMID- 8404440 TI - Comments on "Treatment of diabetic neuropathy with gamma-linolenic acid" by The gamma-Linolenic Multicenter Trial Group. PMID- 8404441 TI - Early referral of children with IDDM to a specialized program correlates with better long-term metabolic control. PMID- 8404442 TI - Intriguing efficacy of Belgian conventional therapy? PMID- 8404443 TI - What level of HbA1c can be achieved in young diabetic patients beyond the honeymoon period? PMID- 8404444 TI - The response of GHb to stepwise plasma glucose change over time in diabetic patients. PMID- 8404445 TI - Thyroid disease in Chinese children with IDDM. PMID- 8404446 TI - Autoimmune thyroid disease in Chinese children with IDDM. PMID- 8404447 TI - Medicare's proposed cutbacks on durable goods stir controversy. PMID- 8404448 TI - Abstracts of the 33rd Meeting of The Association for Eye Research including the annual meeting of the European Club for Ocular Fine Structure. Aachen, Germany, 10-13 June 1992. PMID- 8404449 TI - The importance of diabetes and impaired glucose tolerance in developing countries. PMID- 8404450 TI - Glucose and insulin responses to intravenous glucose challenge in relatives of Nigerian patients with non-insulin-dependent diabetes mellitus. AB - We analysed blood insulin and glucose concentrations before and during frequently sampled intravenous glucose tolerance tests (FSIGT) in 2 groups of Nigerian subjects: (A) Control group (n = 18), without a positive family history of diabetes mellitus, and (B) Experimental group (n = 16), comprising age-, sex- and body mass-matched first-degree relatives of patients with non-insulin-dependent diabetes mellitus (NIDDM). In comparison with Group A subjects, those in Group B had: (i) higher fasting plasma glucose level (mean +/- S.E.M. 4.1 +/- 0.1 vs. 3.8 +/- 0.11 mmol/l, P < 0.05); (ii) similar fasting serum insulin levels (6.7 +/- 5.0 vs. 5.8 +/- 5.6 mU/l, P = NS); (iii) lower mean incremental area under the first-phase (t = 0-10 min) post-glucose challenge insulin curve (376.9 +/- 8.8 vs. 435.6 +/- 5.6 mU/min l-1, P < 0.05); (iv) increased incremental area under the second-phase (t = 10-182 min) post-glucose challenge insulin curve (432.9 +/- 11.5 vs. 161.3 +/- 8.7 mU/min l-1, P < 0.05); (v) reduced KG rate constant of glucose elimination (0.97 +/- 0.12 vs. 1.41 +/- 0.12%/min, P < 0.05). These results suggest that the subjects with a positive family history of NIDDM have a reduced beta-cell insulin secretory reserve (from reduced first-phase insulin response), tendency to rebound hyperinsulinemia during the latter phase of the insulin secretory response, a degree of tissue insulin insensitivity (as evident from high fasting plasma glucose despite similar insulin levels) and a diminished glucose disposal rate, in comparison with subjects without a family history of NIDDM. These features predict subsequent development of diabetes and suggest that as in Caucasians, first-degree relatives of Nigerian patients with NIDDM are at greater risk for future development of the disease. PMID- 8404451 TI - A new diabetes model induced by neonatal alloxan treatment in rats. AB - Rats treated with streptozotocin (STZ) during the neonatal period have been used as a model of non-insulin-dependent diabetes mellitus. The present study was designed to produce another diabetes model by substituting alloxan for STZ. Male Sprague-Dawley rats of 2, 4 or 6 days of age were injected intraperitoneally with 200 mg/kg of alloxan monohydrate after 16 h fast. Control rats received vehicle alone at 6 days of age. Non-fasting plasma glucose levels in alloxan-treated rats significantly increased after 8 weeks as compared with control, as the age of alloxan treatment advanced (6.6 +/- 0.2 (S.E.M.) mM in control, 8.3 +/- 0.3 mM in 2 days, P < 0.05, 9.8 +/- 0.9 mM in 4 days, P < 0.05, 17.1 +/- 3.5 mM in 6 days, P < 0.05). For the long-term observation, alloxan-treated rats were divided into mild and severe diabetes groups. Hyperglycemia persisted in both groups until 52 weeks (6.5 +/- 0.1 mM in control, 10.3 +/- 0.7 mM in mild diabetes group, 25.3 +/ 3.6 mM in severe group), but significant albuminuria developed only in severe diabetes group. The diabetogenicity of alloxan rapidly increased during the neonatal period, and the neonatal alloxan diabetes model may be useful for studying chronic diabetic complications. PMID- 8404452 TI - Zinc deficiency exaggerates diabetic osteoporosis. AB - Streptozotocin diabetic rats showed an increase of bone fragility (11.9 +/- 2.1 kg/cm2 vs. 16.8 +/- 2.0, P < 0.005) which was normalized by insulin treatment (18.3 +/- 4.2), indicating that osteoporosis was induced in diabetic rats. The rats were fed a zinc-deficient diet (0.16 mg/100 g) or a control diet (5.2 mg/100 g). This mild zinc-deficient diet did not lower the serum zinc level. The cortical bone of diabetic rats was shown to be markedly thinner by microscopic examination of ground cross-sections of the tibia. Zinc deficiency induced a reduction in the calcium content of diabetic bone when compared with the rats on a control diet. Urinary excretion of calcium and phosphorus was significantly increased in diabetic rats, and increased further when the rats were fed a zinc deficient diet. Moreover. the bone calcium and phosphorus concentrations were significantly lower in these animals. These changes in the zinc-deficiency rats were not reversed by insulin treatment. Our findings suggest that osteoporosis in the diabetic rats was due to thinning of the bone cortex secondary to mineral loss and can be reversed by insulin treatment, and that these skeletal changes are greatly enhanced by mild zinc deficiency. In addition the effects of zinc deficiency cannot be completely reversed by insulin treatment. PMID- 8404453 TI - Postprandial blood glucose and its relation to diabetic gastroparesis--a comparison of two methods. AB - Delayed gastric emptying is known as an important organic cause for brittle diabetes. We proposed the interval from the start of a meal to the rise in blood glucose, defined as blood glucose latency (T BG) as an index for gastric emptying and a non-invasive test for diabetic gastropathy. In order to validate this test we compared it in 22 type 1 diabetic patients with an established scintigraphic method for the measurement of gastric half-emptying time (T1/2) and found the following correlation: T BG = 4.4 + 0.162 x T1/2; r 0.79, P < 0.001. We therefore suggest measuring the blood glucose latency as a simple non-invasive screening method. PMID- 8404454 TI - Study of the rate of early glucose disappearance following insulin injection: insulin sensitivity index. AB - The euglycaemic hyperinsulinaemic glucose clamp is usually considered as the reference technique to evaluate insulin sensitivity. As it is an expensive and time-consuming tool, we therefore tried to validate a simple insulin tolerance test (ITT) (IV bolus of 0.1 IU/kg of regular insulin, with glucose sampling at 5, 0, 3, 5, 7, 10 and 15 min) and to demonstrate its usefulness. Insulin sensitivity was measured by DG/G0 ratio (G0 = initial glycaemia, DG is the variation between G0 and the glycaemia obtained at 15 min by the calculation of the regression plot). We confirmed the existence of a correlation between the glucose uptake (mg/kg per min) evaluated by glucose clamp and the DG/G0 index (r = 0.9, P < 0.01). There was no stimulation of hormonal counter regulation during the test. The ITT was significantly correlated both with fasting insulin (r = 0.43, P < 0.01), and post-glucose load insulin concentration (r = -0.67, P < 0.01); each measurement expressing insulin sensitivity. Four groups of patients with different insulin sensitivity: controls, NIDDM, gynoid and android obese subjects, were clearly separated by ITT. We showed that fasting glycaemia and DG/G0 were correlated (y = 2.63/x - 0.093; r = 0.82, P < 0.01). These results suggest that ITT could be an easy, quick and low cost method to evaluate insulin resistance in clinical practice and epidemiological studies. PMID- 8404455 TI - Poor cognitive task performance of insulin-dependent diabetic children (6-12 years) in India. AB - Cognitive function was assessed in 22 diabetic and 39 healthy children, all in the age group of 6-12 years. Wechsler's coding, digit span test and Raven's colored progressive matrices were included in the battery of tests. The diabetic children underscored on all these tasks compared to the healthy children. The raw scores (mean +/- S.D.), of the diabetic and healthy children were: Wechsler's coding: 17.4 +/- 6.48, 37.4 +/- 9.33; digit span: 22.0 +/- 8.8, 44.1 +/- 7.67 and on Raven's A+AB+B: 16.2 +/- 3.5. 24.1 +/- 6.26; P < 0.001 in each instance, respectively. Duration of diabetes did not correlate with any of the test scores. No significant differences were noted between the early onset (IDDM starting before the age of 5 years) and late onset (IDDM starting after 5 years) diabetic children in their performance. We attribute the very low scores in our diabetic children to the psycho-social factors in addition to the metabolic control. CONCLUSIONS: (1) specific cognitive dysfunction is present in children with IDDM compared to the healthy children; (2) duration of diabetes did not correlate with cognitive function scores; (3) IDDM manifesting after the age of 5 years also had hitherto unknown detrimental effects on the cognitive process. PMID- 8404456 TI - Cooperation between parents in caring for diabetic children: relations to metabolic control and parents' field-dependence-independence. AB - Aspects of parental interaction were assessed in 20 families with diabetic, insulin-dependent children, using hour-long video-taped interviews, the children being in optimal (O, n = 10) or poor (P, n = 10) metabolic control and showing optimal or poor psychological adaptation. In comparison with the O-group parents, the P-group parents were less appreciative of one another, were less congruent in their attitudes to diabetes care, and appeared not to respect their childrens' independence and integrity; the mothers were discontented with the support given them by their husbands; the children assumed less responsibility for managing their diabetes and seemed less confident during the interview. The results are interpreted in light of an earlier finding that the P-group fathers are more field-dependent (FD) than their wives while the opposite is true for the O-group fathers. With reference to evidence from the cognitive style literature, we suggest that the relatively FD P-group fathers have difficulties in acting as autonomous sources of support to their wives resulting in marital discord and a delayed transition from maternal to self care in their children. PMID- 8404457 TI - Utilisation of health services by aboriginal Australians with diabetes. AB - Diabetes is a major public health problem for Aboriginal Australians. We wished to determine the extent and pattern of health service utilisation by Aboriginal people with diabetes in central Australia. Medical records of all Aboriginal people known to have diabetes (n = 374), identified by a previous study, were examined for attendance to health services in central Australia. All had non insulin-dependent diabetes. Between January 1984 and December 1986, Aboriginal adults with diabetes were admitted to hospital on 694 occasions, accounting for 10.8% of adult Aboriginal admissions. The crude admission rates were 0.78 and 0.84 per diabetes-year for men and women, respectively. The age-adjusted relative risks for admission, compared with Aboriginal non-diabetic patients, were 2.93 (95% C.I., 2.62-3.26) for men and 2.46 (95% C.I., 2.28-2.66) for women. If admission for conditions associated with diabetes are excluded, the admission rates were similar for the two groups. Infection was the most common reason for attendance to a health service, representing 41.7% and 39.8% of male and female admissions, and 21.8% and 26.3% of male and female outpatient attendances. Aboriginal patients with diagnosed diabetes suffer high morbidity and contribute disproportionately to health system costs. PMID- 8404458 TI - The proportion of death certificates of diabetic patients that mentioned diabetes in Osaka District, Japan. AB - To determine the accuracy of the stated causes of death in diabetic patients, we analyzed 503 death certificates of diabetic patients who had been followed up for 9.4 years on average at Osaka Medical Center for Adulthood Diseases and had died during the observation period from 1960 to 1984. Of the certificates examined, only 213 (42.4%) mentioned diabetes, either as the underlying cause or as a contributory condition. The proportion of certificates that mentioned diabetes was related to the underlying cause of death; it was 18.7% for malignant neoplasms, 40.5% for cerebrovascular disease and 46.9% for disease of the heart, while it was 81.8% for renal disease. The proportion was higher for females (48.7%) than for males (39.5%), and a significant difference was observed between the sexes in the proportion in the case of disease of the heart. The proportion was not related to any significant extent to the year of death or the age at death. However, it was related to fasting plasma glucose (FPG) level and type of treatment. It was 37.8% and 54.4% for patients with FPG < 200 mg/dl and FPG > or = 200 mg/dl, respectively, 24.1% for those treated with diet, 51.4% for those treated with oral hypoglycemic agent and 80.7% for those treated with insulin. As a result, it was found that the proportion of death certificates that mentioned diabetes was related to the accompanying complications and the severity of diabetes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404459 TI - Clinical pharmacokinetics and metabolism of benzimidazole anthelmintics in ruminants. PMID- 8404460 TI - Metabolism and pharmacokinetics of T-2 toxin and related trichothecenes. PMID- 8404461 TI - Cytochromes P450: their active-site structure and mechanism of oxidation. PMID- 8404462 TI - The management of polymyalgia rheumatica and giant cell arteritis. PMID- 8404463 TI - Fluoroquinolones reviewed. PMID- 8404464 TI - [Resuscitation by hospital personnel. Self evaluation and educational status]. PMID- 8404465 TI - [Cardiopulmonary resuscitation]. PMID- 8404466 TI - [Backache and spinal mechanics]. PMID- 8404467 TI - [Rational solutions of diagnostic problems in back disorders]. PMID- 8404468 TI - [Psychogenic lumbar-ischial pain syndrome]. PMID- 8404469 TI - [Osteoporosis]. PMID- 8404470 TI - [Spinal canal stenosis]. PMID- 8404471 TI - [When back pain turns chronic. The importance of psyche, work and treatment]. PMID- 8404472 TI - [Epidemiology of back pain. Introduction to a research project]. PMID- 8404473 TI - [Ergonomic construction of office work places in hospitals]. PMID- 8404474 TI - [Scheuermann's disease; diagnosis and therapy]. PMID- 8404475 TI - [Restenosis after the implantation of Palmaz-Schatz vascular stents in the coronary arteries]. AB - Restenosis rate after successful intracoronary implantation of Palmaz-Schatz stents in 100 patients (92 men, 8 women; mean age 57 +/- 11 years) was quantitatively assessed by angiography performed on average 5.3 +/- 0.3 months after the procedure. Restenosis was defined as a more than 50% decrease in lumen. Data from patients with acute or subacute thrombotic complications were excluded from the analysis. The restenosis rate of the total group was 22%. After placement of only one stent (n = 87) it was 17%, of multiple stents per lesion (n = 13) 54%. Restenosis rate after emergency implantation of a single stent (n = 23) was 17.4%, after elective single stent implantation (n = 64) 17.2%. There was no significant difference regarding treatment of new stenoses (n = 16), and recurrent stenosis (n = 48), namely 12.5% vs 18.8%. The following were risk factors for chronic restenosis after stent implantation: multiple stents (odds ratio [OR] 5.6; 95% confidence interval [CI]: 1.6-19.1); implantation in a small vessel, reference diameter < or = 3.0 mm (OR 6.7, CI 2.4-18.7); and residual stenosis after stent implantation of > 8% (OR 3.1, CI 1.2-8.1). PMID- 8404476 TI - [Fulminant hepatitis caused by disulfiram]. AB - After undergoing withdrawal treatment for alcoholism as an in-patient for one year a 49-year-old woman was started on disulfiram, 250 mg daily, her liver function tests being normal. Except for vitamin B1 she received no further medication. Jaundice developed 13 days after onset of treatment and acute liver failure was diagnosed on the 18th day after a total disulfiram dose of 4.5 g (Quick value < 10%; bilirubin 460 mumol/l; GPT 5099 U/l; GOT 4142 U/l), as well as early renal failure (creatinine 300 mumol/l). An acute viral infection, autoimmune hepatitis and a metabolic liver disease were excluded by biochemical, serological and molecular biology tests. All toxicological tests were negative. The patient died 25 days after the onset of disulfiram treatment in hepatic coma due to a fulminant hepatitis with hepatorenal syndrome. Both a liver biopsy and the autopsy showed the signs of an acute hepatic dystrophy without cirrhosis. The temporal relationship between the disulfiram intake and onset of the illness, the exclusion of other causes of the fulminant hepatitis and the liver histology, which was compatible with a chemical-toxic hepatitis, indicate that this was a case of disulfiram-induced hepatitis. The hepatotoxicity of disulfiram is a very rare idiosyncratic reaction which is often fatal. Disulfiram administration must be discontinued at once if there is a rise in liver enzyme activity or jaundice occurs. PMID- 8404477 TI - [Trichobezoar as a rare cause of ileus of the small intestine]. AB - For several weeks a 15-year-old girl had complained of increasing abdominal pain with vomiting. On admission to hospital the bloated abdomen was diffusely sensitive to pressure and the bowel sounds were high pitched and loud. Erythrocyte sedimentation rate was increased to 23/40 mm and the white cell count to 12,000/microliters. Ultrasound examination revealed an echo-dense area with dorsal echo loss at the gastric side of the pylorus. X-ray films of the stomach showed fluid levels and a soft-tissue mass in the left upper abdomen. At laparotomy a large (12 x 6 cm) trichobezoar was found in the middle of the small intestine and two smaller ones at the pylorus. Subsequently the patient admitted to trichophagia but refused any psychological treatment. PMID- 8404478 TI - [Guidelines for substitution therapy with thrombocytes]. PMID- 8404479 TI - [Chronic hepatitis B virus infection: the mechanisms of cell damage]. PMID- 8404480 TI - [The chronic fatigue syndrome]. PMID- 8404481 TI - [A temporary cardiac pacemaker before major operations?]. PMID- 8404482 TI - [The classification of protein storage diseases]. PMID- 8404483 TI - [Rectus-vaginal hematoma]. PMID- 8404484 TI - [An ulcer in the esophagus with a vascular stump as the cause of bleeding]. PMID- 8404485 TI - [A synthetic vaccine against Plasmodium falciparum]. PMID- 8404486 TI - [Hormonal substitution after menopause]. PMID- 8404487 TI - [Physical training status and cardiovascular mortality]. PMID- 8404488 TI - [Computerized tomography-guided fine needle biopsy. Current perspectives in biopsy diagnosis]. AB - 253 cutting needle biopsies from 240 patients (151 men, 89 women; mean age 68 [23 84] years) were reviewed. The biopsies were taken under computed (CT) tomography guidance from various parts of the body (pancreas: n = 48, liver: n = 32, other upper abdominal organs: n = 5, mediastinum: n = 27, lung: n = 37, kidney: n = 13, adrenals: n = 8, retroperitoneum: n = 35, pelvis: n = 37, pleura: n = 4, chest wall: n = 6). The data were evaluated retrospectively. In 86% of cases meaningful clinical information was obtained in the form of a definite diagnosis (74%) or of a presumptive or differential diagnosis (12%). As a rule it was possible to assess the degree of malignancy and to ascertain the exact nature of the condition; the latter was true not only of malignant but also of benign lesions. In the majority of cases this had important implications. In just under 14% of all cases the target area was missed and no diagnosis could be made. To raise the chances of obtaining a representative biopsy multiple punctures were performed in 44% of the CT-guided biopsies. In 61 cases (24%) the diagnosis was checked against further tissue samples and in four cases the diagnosis based on needle biopsy material had to be revised. In all, only two complications were recorded; both were mild and did not require treatment. In terms of diagnostic value, CT guided fine needle biopsies proved considerably superior to conventional cytological studies of fine needle aspiration material. PMID- 8404489 TI - [Toxic shock syndrome caused by Streptococcus pyogenes]. AB - Four days after being bitten by an insect a 35-year-old woman without any serious underlying disease developed an extensive phlegmonous inflammation of the left eyelid which soon spread to the entire left half of her face. Streptococcus pyogenes serotype M1, which produced the erythrogenic toxin A in vitro, was isolated from two blood cultures. The course of the illness was characterized by high fever, diarrhoea, vomiting, circulatory failure, consumption coagulopathy, abnormal renal functions and a generalized exanthem with desquamation of the skin, exhibiting the full-blown picture of a toxic shock syndrome caused by S. pyogenes. She eventually recovered completely under intensive care involving administration of catecholamines, fresh frozen plasma and antithrombin III substitution, as well as antibiotic treatment with clindamycin (600 mg three times daily), ampicillin/sulbactam (4 g three times daily)--after 3 days replaced by imipenem (0.5 g four times daily)--and gentamycin (80 mg three times daily) for two weeks. Extensive necroses later required plastic surgery to the left eyelid, cheek and temporal region. PMID- 8404490 TI - [Staging laparoscopy in Hodgkin's disease. A valid alternative to staging laparotomy]. AB - A 30-year-old man with Hodgkin's disease, stage IA, had received radiotherapy to the supradiaphragmatic lymph nodes two years ago. During follow-up observations the patient felt well, but sonography revealed three tumours in the spleen. The erythrocyte sedimentation rate was found to be raised to 50/75 mm. Therefore, splenectomy and staging of the other parts of the abdomen for histological clarification and deciding on further therapeutic action were indicated. Thus splenectomy with bilateral liver wedge resections were performed laparoscopically, together with excision of mesenteric, paraaortic and para-iliac lymph nodes. The patient was discharged home on the 4th postoperative day. Histological examination revealed stage IIIE Hodgkin's disease. This case demonstrates that in favourable conditions staging laparoscopy can obtain the same information as conventional staging laparotomy, but it is much better tolerated by the patient. PMID- 8404491 TI - [Metabolic syndrome. A new challenge for preventive medicine]. PMID- 8404492 TI - [Essential thrombocythemia. Clinical significance, diagnosis and therapy]. PMID- 8404493 TI - [Ambulatory surgery--day surgery--short-stay surgery]. PMID- 8404494 TI - [Filters in blood transfusions]. PMID- 8404495 TI - [Movement therapy and myocardial ischemia]. PMID- 8404496 TI - [Annual test for occult blood in feces and colonic cancer mortality]. PMID- 8404497 TI - [Color-coded duplex ultrasonography in the diagnosis of renal artery stenosis]. AB - To diagnose possible renal artery stenosis in 120 patients (36 women, 84 men; mean age 57.3 [21-84] years) colour duplex sonography (CDS) of the renal arterial tree was performed before 53 intraarterial angiographies and 67 central venous digital subtraction angiographies. The criterion of stenosis was a maximal flow velocity of > 180 cm/s in the colour duplex sonogram and a diameter reduction of > 50% in the angiogram. CDS demonstrated the renal artery bed in 209 of 247 renal arteries (85%). 84 of the 209 had regional maximal velocities of > 180 cm/s. The sensitivity and specificity of CDS when comparing it with both angiographic methods (in 185 cases that could be evaluated, in 74 of them with abnormal results) were 91 and 88.2%, respectively. Comparing CDS results with those by intraarterial angiography alone (88 cases, with 51 abnormal findings), sensitivity and specificity were 92.1 and 91.8%, respectively. These results demonstrate that CDS can reliably demonstrate flow obstructions in the renal arterial tree and can thus be recommended for the diagnosis of renovascular hypertension. PMID- 8404498 TI - [A severe course of leptospirosis with acute kidney failure and extensive icterus (Weil disease)]. AB - A 77-year-old man developed a fever up to 38.4 degrees C, with diarrhoea, acute renal failure (creatinine up to 8.7 mg/dl; urea up to 308 mg/dl) and marked jaundice (total bilirubin up to 24.3 mg/dl). In addition there was thrombocytopenia, conjunctivitis and epistaxis, as well as cerebral symptoms with somnolence and general slowing up. At first he was thought to have cholangitis resulting from previously diagnosed gall-stones, and he was therefore treated with ampicillin, 2 g two times daily, and metronidazole, 0.5 g two times daily. The fewer regressed, but the renal failure required haemodialysis and haemofiltration under strict fluid control. Endoscopy excluded obstructive jaundice, but a suspicion of inflammatory liver disease or possibly cirrhosis was raised in the differential diagnosis. Serology revealed an increased titre for Leptospira interrogans var. sejroe (1:200, later 1:1600). Liver biopsy finding was compatible with the diagnosis of leptospirosis. Because of the high inflammatory activity in the liver, 2 mega units of penicillin G were administered three times daily for six days. Gradually the renal functions and jaundice improved and, on discharge on the 36th day, the patient was again in generally good health, although creatinine and bilirubin values were still slightly elevated (1.7 mg/dl each). PMID- 8404499 TI - [Pathology and clinical aspects of peripheral neuroectodermal tumors]. AB - A 36-year-old man developed progressive pain in the left thorax radiating to the shoulder, associated with weight loss as well as axillary and supraclavicular lymph-node swellings, and a left Horner's syndrome. The radiological diagnosis was Pancoast tumour. Computed tomography of the thorax revealed that the tumour had almost completely infiltrated the left lung and hilus. Sonography demonstrated metastases in the liver and retroperitoneum. Histological examination of an excised axillary lymph-node metastasis showed an undifferentiated small-cell tumour. Immunohistochemical tests revealed expression of protein S 100 and neurofilamental proteins, i.e. evidence of a peripheral malignant neuroectodermal tumour. The tumour also expressed vimentin. Treatment consisted of two chemotherapy cycles according to the EVAIA scheme (daily three times 235 mg etoposide, 2 mg vincristine, three times 0.8 mg actinomycin D, three times 3.2 g ifosfamide, three times 30 mg doxorubicin, and three times 12 mg dexamethasone), without any effect on the rapid progression of the disease. He died 17 weeks after the diagnosis had been made. PMID- 8404500 TI - [Diagnosis of Raynaud's syndrome]. PMID- 8404501 TI - [Holt-Oram syndrome]. PMID- 8404502 TI - [Acute mountain sickness and high altitude pulmonary edema]. PMID- 8404503 TI - [Influenza infection and antibiotic therapy]. PMID- 8404504 TI - [Severe tetraplegia in chronic lymphatic leukemia]. PMID- 8404505 TI - [Maternal antigen as a risk factor in rheumatoid arthritis]. PMID- 8404506 TI - [Granulocyte function in myelodysplastic syndrome before and after low-dose cytarabine therapy]. AB - Granulocytic phagocytosis ability (phagocytosis index in peripheral blood and in skin chamber exudate) and in vivo chemotaxis (skin chamber leukocyte mobilisation) were investigated in 22 patients with myelodysplastic syndrome (19 men, three women, mean age 66.1 [46-82] years). The control group consisted of 20 young, healthy subjects (13 men, seven women, mean age 25 [22-27] years). The studies were repeated in twelve of the patients after 10 days of low-dose cytarabine therapy (10 mg twice daily subcutaneously). Before treatment, total leukocyte mobilisation (TLM) after 24 hours (3.43 +/- 0.6 G/l.cm2) and phagocytosis index (PI(blood): 33.9 +/- 16.6%, PI(skin) chamber: 34.1 +/- 13.1%) were significantly lower (P < 0.05) than in controls (TLM: 6.7 +/- 0.78 G/l.cm2, PI(blood): 59.6 +/- 17.8%, PI(skin) chamber: 37.4 +/- 18.1%). Following treatment for 10 days, the values were still depressed or had even worsened (TLM: 3.09 +/- 0.97 G/l.cm2; PI(blood): 35.6 +/- 24.1%, PI(skin) chamber: 19.9 +/- 8.6%; all P < 0.05). Investigation of granulocytic function in myelodysplastic syndrome may reveal evidence of reduced immune protection by these cells against infection. Low-dose cytarabine therapy does not seem to affect cell differentiation. PMID- 8404507 TI - [Epithelioid leiomyosarcoma of the stomach with hemorrhagic ascites]. AB - In two men (69 and 65 years old) a gastric epithelioid leiomyosarcoma more than 25 cm in diameter first manifested itself by an increase in abdominal girth and ascites. In case 1, a cylindrical mass was palpable in the abdomen increasing in size over two years. Sonography and computed tomography showed a large cystic tumour which could not be related to any one organ. Cytology of the ascites fluid did not reveal any tumour cells. At operation a pedunculated tumour, 29 x 20 x 9 cm, was found, originating from the stomach. Six litres of ascites fluid were aspirated. There were no detectable abdominal metastases. The patient died nearly 4 years later after the tumour had been resected, three further laparotomies had to be performed because of extensive metastases to peritoneum and liver from the leiomyosarcoma. In case 2, no abdominal mass was palpable, but the patient's girth had increased over the last year. Sonography revealed a cystic tumour, about 30 cm in diameter. Examination of haemorrhagic ascites fluid (700 ml) did not show any tumour cells. The tumour, originating from the stomach wall, was excised (no metastases were seen). The patient remains symptom-free after 35 months. PMID- 8404508 TI - [Ventricular fibrillation and silent myocardial ischemia in a patient without anatomic heart disease]. AB - A 44-year-old man, apparently without heart disease, suddenly collapsed with loss of consciousness. Ventricular fibrillation was documented when an external defibrillator was connected. After cardiopulmonary resuscitation and intubation he quickly regained consciousness. Clinical examination together with echocardiography, coronary angiography and electrophysiological tests discovered no abnormalities. Biochemical tests were normal except for slightly abnormal liver functions. Several long-term ECG recordings documented asymptomatic S-T elevations. During one such episode there occurred a polymorphous ventricular tachycardia of brief duration. Because the S-T elevations persisted, despite the administration of gallopamil (50 mg twice daily for one week), a defibrillator was implanted. Gallopamil was then discontinued. Long-term ECG monitoring subsequently revealed four episodes of marked S-T elevations, three of which accompanied by ventricular arrhythmias. After resuming gallopamil, now at a dose of 50 mg three times daily, further ECG monitoring showed no abnormalities. PMID- 8404509 TI - [Therapy of Raynaud's syndrome]. PMID- 8404510 TI - [The clinical significance of penicillin-resistant pneumococci]. PMID- 8404511 TI - [Advertising by way of physician's letter-head. Decision by the Federal Constitutional Court of 4-21=1993]. PMID- 8404512 TI - [Radiation injury caused by exposure to ultraviolet and infrared light]. PMID- 8404513 TI - [Arrhythmia caused by a too rapid central venous administration of cefotaxime]. PMID- 8404514 TI - [Bases of and developments in homeopathy]. PMID- 8404515 TI - [Prothrombin fragment F1 + 2]. PMID- 8404516 TI - [Oral contraception and cerebral embolisms]. PMID- 8404517 TI - [80 year constitution of the rectorate of the School of Veterinary Medicine Hannover]. AB - On the July 18th 1993, the School of Veterinary Medicine existed for 215 years. The School named royal "Rossarzneischule" was founded under the direction of the "Oberhofrossarzt" JOHANN ADAM KERSTING by means of the royal order of George III, King of England and Duke of Hannover, in 1778. Shortly afterwards it had expanded to the royal "Thierarznei-Schule" and the School of Veterinary Medicine obtained its present name in 1887. A constitution of the rectorate was obtained in March 31 by royal decree. The inauguration of the first rector, BERNHARD MALKMUS, took place on June 16th in 1913. Up to now, 36 rectors held office at this university. An epilogue of this constitution indicates the early history from 1887-1913. A chronology of the rectors is shown at the end of this paper. PMID- 8404518 TI - [The content of ascorbic acid in tissues of swine fetuses and newborn piglets and in the blood plasma and tissue of swine of different ages with regard to the effect of the restriction of suckling time]. AB - In 55, 75, 95 and 110 days old pig-fetuses as well as in newborn piglets the weight of different tissues and their content of ascorbic acid were analysed. The time for suckling of the piglets of 2 sows was restricted to 12 hours per day (= group A); the piglets of 2 sows were used as a control (= group B). The restriction of the milk intake did not influence the content of the blood plasma and of the tissues in ascorbic acid. After weaning the pigs of group A had a compensatory growth and on the 214th day after birth the same slaughter mass as that of the group B. There was no difference in the content of ascorbic acid of the tissues of the 2 groups of pigs. The significance of changes in the content of ascorbic acid in the tissues during the fetal and the postnatal growth is described. PMID- 8404519 TI - Naloxone affects gastrointestinal functions and behaviour in horses. AB - Clinical effects of the opioid antagonist naloxone were investigated in healthy horses. Naloxone caused a transient increase in the frequency of defecations, a softening of the faeces and alterations in the intensity of abdominal borborygmi. Total serum protein concentrations decreased. Behavioural changes like frequent yawning and flehmen occurred, heart rate decreased and respiratory rate tended to increase but no clinical signs of distress or pain were observed. It can be concluded that in healthy animals naloxone had only minor side-effects and could be used to investigate the physiological functions of endogenous opioids in horses. PMID- 8404520 TI - Comparative haemodynamic alterations induced by pipecuronium and pancuronium. AB - Haemodynamic effects of pipecuronium bromide (Pi.) and pancuronium bromide (Pa.) were studied on isolated rabbit's heart, guinea pig's tracheal chain as well as the blood pressure in pentobarbital anaesthetized dogs. Pi. induced negative inotropic and chronotropic effects on the isolated rabbit's heart especially in lower concentrations. However, higher concentrations provoked two opposite effects, negative chronotropic and positive inotropic activity. In addition, Pa. in lower concentrations caused positive inotropic and negative chronotropic activity, while higher concentrations induced negative inotropic and chronotropic activity. Cardioinhibitory actions of both tested drugs are not due to either cholinergic or beta 1-adrenergic blocking effect but it may be due to nicotine like activity. In anaesthetized dogs, i.v. injections of both tested drugs produced a transient decrease in systolic and diastolic pressure in doses above the therapeutic level. This effect may be referred to the partial ganglion blocking effect of both tested drugs. PMID- 8404521 TI - [The effect of husbandry, management and stall climate on lung changes in swine]. AB - 21 swine fattening units (17 closed and 4 open swine herds), their pigs suffering from chronic respiratory disease since a long time were investigated concerning fattening period, morbidity, mortality, lung lesions by slaughtered swine, microorganisms involved, husbandry and management conditions, stall climate (indoor temperature, relative air humidity, contents of NH3, CO2, H2S, air microorganisms and dust). We estimated annual fattening periods of 137.4 +/- 6.8 days, morbidity of 49.1 +/- 12.2% and mortality of 6.8 +/- 3.7%. About 55-96% of lungs of slaughtered pigs showed pneumonic lesions in different degrees. Bacteriological examination of such lesions revealed up to 9 and not less than 3 different bacteria species per unit. Continuous producing systems were used by 19 units, only 2 units were managed by all in all out system. Pigs of 70 kg weight have air spaces of 1.7-3.1 m3, indoor temperature were between 21-26 C degrees and relative air humidity between 83.9 +/- 13.7%. Concentration of NH3 was found between 3-35 ppm, of CO2 between 500-3500 ppm, the numerical content of bacteria was within 900-1620 cfu (colony forming units) and of fungi between 2-80 cfu per litre air. There were also E. coli with and without haemolysis, Klebsiella pneumoniae, beta-haemolyzing streptococci and staphylococci in stall air. 2 swine houses gave 3-5, 15 houses 12-50 and 4 houses more than 50 cfu/cm2 blood agar plates exposed for 1 minute to the stable air (sedimentation method to judge stall dust). Animals of the later mentioned 4 houses showed the highest incidence of respiratory disorders (> 50% of pigs).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404522 TI - [Fearfulness in animals--a zoological and forensic problem]. AB - The recognition and the assessment of anxiety and fear in animals is discussed regarding zoological, forensic and ethical aspects. Following some definitions it is shown that fear, anxiety and other emotions are generally assessed by analogous reasoning and "Du-Evidenz". This causes scientific and forensic problems which are discussed. Towards a more adequate assessment of emotions in animals a zoological approach based on integrated ethological and stress physiological theories and methods is proposed. PMID- 8404523 TI - [Hedgehog care?]. AB - The european hedgehog (Erinaceus europaeus) is covered by species protection laws; it is also covered by the animal protection laws. It is not permitted to capture it, to injure it, to kill it or to want only disturb it. As a rule it is also not permitted to be held in captivity, to be sold or put out to show. Disregard of the protection laws can be punished with fines or prison sentences. Irrespective of the legal point of view, according to todays understanding wild animals belong in free nature, so that they can live a style of life suitable for their species. PMID- 8404524 TI - Antibody prevalence of hog cholera, bovine viral diarrhoea and Aujeszky's disease virus in wild boars in northern Germany. AB - During the hunting season 1990/1991 a total of 841 blood samples was collected from shot wild boar corresponding to about 2.11% of the total hunting bag in Lower Saxony. All the sera were screened for neutralizing antibodies (nAb) to hog cholera virus (HCV) and bovine viral diarrhoea virus (BVDV) by direct neutralizing peroxidase linked antibody (NPLA) assay. For the detection of antibodies (Ab) against HCV a Complex Trapping Blocking (CTB) ELISA was used. Cytotoxic sera were retested using an indirect immunoperoxidase technique. Additionally all sera were tested for antibodies to Aujeszky's disease virus (ADV) in a commercial indirect ELISA and 722 sera for ADV nAb in a virus neutralization test (VNT) with and without the addition of guinea pig complement. Screening of 841 sera yielded six sera neutralizing HCV strain ALFORT/187 and two sera with a positive CTB-ELISA reaction. A total of seven sera not identical with the HCV seropositive sera yielded BVDV neutralizing antibodies. The majority of HCV nAb positive samples were found in a region close to the former border to the German Democratic Republic (GDR) west of the State of Sachsen-Anhalt. Screening for ADV detected five sera positive in ELISA amongst 12 sera yielding nAb against ADV. Positive samples were obtained in the regions of Brunswick, Luneburg and Hannover. PMID- 8404525 TI - Regulation of development and differentiation by the extracellular matrix. PMID- 8404526 TI - Cross regulation of decapentaplegic and Ultrabithorax transcription in the embryonic visceral mesoderm of Drosophila. AB - The Drosophila decapentaplegic gene (dpp) encodes a TGF-beta family member involved in signal transduction during embryonic midgut formation. The shortvein (shv) class of cis-regulatory dpp mutants disrupt expression in parasegments 4 and 7 (ps4 and ps7) of the embryonic visceral mesoderm (VM) surrounding the gut and cause abnormalities in gut morphogenesis. We demonstrate that cis-regulatory elements directing expression in ps4 and ps7 are separable and identify DNA fragments that generate ps4 and ps7 expression patterns using reporter gene constructs. dpp reporter gene expression in both ps4 and ps7 is autoregulated as it requires endogenous dpp+ activity. Reporter gene ps7 expression requires the wild-type action of Ultra-bithorax (Ubx), and abdominal-A. Furthermore, the expression of certain Ubx reporter genes is coincident with dpp in the VM. Both the mis-expression of Ubx reporter genes in the developing gastric caecae at ps4 and its normal expression in ps7 are dependent upon endogenous dpp+ activity. We conclude that dpp both responds to and regulates Ubx in ps7 of the visceral mesoderm and that Ubx autoregulation within this tissue may be indirect as it requires more components than have previously been thought. PMID- 8404527 TI - Analysis of genetic mosaics in developing and adult Drosophila tissues. AB - We have constructed a series of strains to facilitate the generation and analysis of clones of genetically distinct cells in developing and adult tissues of Drosophila. Each of these strains carries an FRT element, the target for the yeast FLP recombinase, near the base of a major chromosome arm, as well as a gratuitous cell-autonomous marker. Novel markers that carry epitope tags and that are localized to either the cell nucleus or cell membrane have been generated. As a demonstration of how these strains can be used to study a particular gene, we have analyzed the developmental role of the Drosophila EGF receptor homolog. Moreover, we have shown that these strains can be utilized to identify new mutations in mosaic animals in an efficient and unbiased way, thereby providing an unprecedented opportunity to perform systematic genetic screens for mutations affecting many biological processes. PMID- 8404528 TI - Integrin alpha subunit mRNAs are differentially expressed in early Xenopus embryos. AB - Adhesion of cells to extracellular matrix proteins is mediated, in large part, by transmembrane receptors of the integrin family. The identification of specific integrins expressed in early embryos is an important first step to understanding the roles of these receptors in developmental processes. We have used polymerase chain reaction methods and degenerate oligodeoxynucleotide primers to identify and clone Xenopus integrin alpha subunits from neurula-stage (stage 17) cDNA. Partial cDNAs encoding integrin subunits alpha 2, alpha 3, alpha 4, alpha 5, alpha 6 and an alpha IIb-related subunit were cloned and used to investigate integrin mRNA expression in early embryos by RNase protection assay and whole mount in situ hybridization methods. Considerable integrin diversity is apparent early in development with integrins alpha 2, alpha 3, alpha 4, alpha 5 and alpha 6 each expressed by the end of gastrulation. Both alpha 3 and alpha 5 are expressed as maternal mRNAs. Zygotic expression of alpha 2, alpha 3, alpha 4 and alpha 6 transcripts begins during gastrulation. Integrin alpha 5 is expressed at relatively high levels during cleavage, blastula and gastrula stages suggesting that it may represent the major integrin expressed in the early embryo. We demonstrated previously that integrin beta 1 protein synthesis remains constant following induction of stage 8 animal cap cells with activin (Smith, J. C., Symes, K., Hynes, R. O. and DeSimone, D. W. (1990) Development 108, 289-298.). Here we report that integrin alpha 3, alpha 4 and alpha 6 mRNA levels increase following induction with 10 U/ml activin-A whereas alpha 5, beta 1 and beta 3 mRNA levels remain unchanged. Whole-mount in situ hybridization reveals that alpha 3 mRNAs are expressed by cells of the involuting mesoderm in the dorsal lip region of early gastrulae. As gastrulation proceeds, alpha 3 expression is localized to a stripe of presumptive notochordal cells along the dorsal midline. In neurulae, alpha 3 mRNA is highly expressed in the notochord but becomes progressively more restricted to the caudalmost portion of this tissue as development proceeds from tailbud to tadpole stages. In addition, alpha 3 is expressed in the forebrain region of later stage embryos. These data suggest that integrin-mediated adhesion may be involved in the process of mesoderm involution at gastrulation and the organization of tissues during embryogenesis. PMID- 8404529 TI - Development of Drosophila wing sensory neurons in mutants with missing or modified cell surface molecules. AB - The neurons of the sensory receptors on the wing of Drosophila melanogaster have highly characteristic axon projections in the central nervous system (CNS). The morphology of these projections was studied in flies bearing mutations that affect cell surface molecules thought to be important in axon guidance. The animals used were mutant for the fasciclinI (fasI), fasciclinII (fasII), fasciclinIII (fasIII) and neurally altered carbohydrate (nac) genes. Axon populations were visualized by staining with DiI and light-reacting the dye with diaminobenzidine to yield permanent preparations. The fasI, fasII and fasIII mutants as well as the nac mutant display altered axonal trajectories in the CNS. One phenotype seen in fasII mutants and in animals mutant for both fasI and fasIII was extra branching within the axon projection pattern. A second phenotype observed was a reduction or complete loss of one of the tracts, apparently due to the axons shifting to a neighboring tract. This was seen in the most extreme form in nac mutants and to a lesser degree in fasIII mutants. To determine if the mutations discussed here affected axon guidance, wing discs were analyzed using the antibody 22C10 to label sensory neurons in the wing during metamorphosis. Both misrouting of axons and the appearance of ectopic neurons in the wing were observed. In the fasI:fasIII, the fasII and the nac mutants, there was misrouting of sensory axons in the developing wing. In addition, the fasII and nac mutants displayed ectopic sensory neurons in the wing. This implies that the cell surface molecules missing (fasciclins) or modified (by the nac gene product), in these mutants may play a role in both neurogenesis and axon guidance. PMID- 8404530 TI - Mesodermal cell interactions in the sea urchin embryo: properties of skeletogenic secondary mesenchyme cells. AB - An interaction between the two principal populations of mesodermal cells in the sea urchin embryo, primary and secondary mesenchyme cells (PMCs and SMCs, respectively), regulates SMC fates and the process of skeletogenesis. In the undisturbed embryo, skeletal elements are produced exclusively by PMCs. Certain SMCs also have the ability to express a skeletogenic phenotype; however, signals transmitted by the PMCs direct these cells into alternative developmental pathways. In this study, a combination of fluorescent cell-labeling methods, embryo microsurgery and cell-specific molecular markers have been used to study the lineage, numbers, normal fate(s) and developmental potential of the skeletogenic SMCs. Previous fate-mapping studies have shown that SMCs are derived from the veg2 layer of blastomeres of the 64-cell-stage embryo and from the small micromeres. By specifically labeling the small micromeres with 5 bromodeoxyuridine, we demonstrate that descendants of these cells do not participate in skeletogenesis in PMC-depleted larvae, even though they are the closest lineal relatives of PMCs. Skeletogenic SMCs are therefore derived exclusively from the veg2 blastomeres. Because the SMCs are a heterogeneous population of cells, we have sought to gain information concerning the normal fate(s) of skeletogenic SMCs by determining whether specific cell types are reduced or absent in PMC(-) larvae. Of the four known SMC derivatives: pigment cells, blastocoelar (basal) cells, muscle cells and coelomic pouch cells, only pigment cells show a major reduction (> 50%) in number following SMC skeletogenesis. We therefore propose that the PMC-derived signal regulates a developmental switch, directing SMCs to adopt a pigment cell phenotype instead of a default (skeletogenic) fate. Ablation of SMCs at the late gastrula stage does not result in the recruitment of any additional skeletogenic cells, demonstrating that, by this stage, the number of SMCs with skeletogenic potential is restricted to 60-70 cells. Previous studies showed that during their switch to a skeletogenic fate, SMCs alter their migratory behavior and cell surface properties. In this study, we demonstrate that during conversion, SMCs become insensitive to the PMC-derived signal, while at the same time they acquire PMC specific signaling properties. PMID- 8404531 TI - Gene expression during imaginal disc regeneration detected using enhancer sensitive P-elements. AB - When imaginal disc fragments from Drosophila are cultured in adult female hosts, they either duplicate the part of the pattern specified by the fate map, or regenerate to replace the missing part. The new tissue is added by proliferation of a small number of cells from the cut edge, brought together when the wound heals to form a regeneration blastema. Specification of the new pattern has been explained by assuming interactions among cells of different positional value in the regeneration blastema. In order to identify genes which might mediate these events, we screened over eight hundred independently isolated autosomal insertions of an enhancer-sensitive P-element, for altered lac-z expression in regenerating discs following cell death induced by a temperature-sensitive cell lethal mutation. Two further screens divided the positive lines into four groups based on appropriate timing of the lac-z response in the cell-lethal mutant background and the expected response to an alternate source of cell death. Expression in wing disc fragments cultured in vivo was most frequent in the target class defined by the screens. In this direct test, lac-z expression was found in 23 lines and in most cases was spatially and temporally correlated with the formation of the regeneration blastema. Our results suggest a very substantial transcriptional response during the early stages of imaginal disc regeneration. lac-z expression in control imaginal discs, embryos and adult ovaries of the positive lines was also assayed. The selected insertions included: a small class expressed only in discs undergoing regeneration and apparently not at any other stage, possibly representing genes active exclusively in regeneration; a larger class expressed in the embryo or during oogenesis, but not normally in imaginal discs, as expected for functions recruited from earlier stages of the developmental program; and finally a class with spatially patterned expression in normal discs. This class included several insertions with expression associated with compartment boundaries, including one at the decapentaplegic (dpp), and one at the crumbs (crb) locus, a growth factor homologue, and an EGF-repeat gene respectively. Some of the expression patterns observed in cultured disc fragments provide evidence for cell communication in the regeneration blastema. PMID- 8404532 TI - Developmental regulation of transcription of a novel prespore-specific gene (Dp87) in Dictyostelium discoideum. AB - The Dp87 is a novel prespore specific gene of Dictyostelium discoideum which has a long open reading frame of 555 amino acids. The entire amino acid sequence had low but significant homology to the spore coat proteins, SP96 and SP70, of this organism. When a chimeric gene, containing a 1380 bp of the 5' upstream region of this gene fused with CAT gene, as reporter, was introduced into cells of this organism, it was expressed only in prespore cells of the slug. Transformation experiments, using chimeric genes, containing a series of 5' deletions of the upstream region, showed that -447 bp to -357 bp is an important cis-acting regulatory region for transcription. A nuclear factor(s) that specifically bind to this cis-acting region were detected from slug cell nuclei. Transformation experiments using a chimeric gene consisting of the 5' region between -666 bp and +149 bp of this gene, a beta-galactosidase reporter and an actin 8 terminator, showed that the reporter gene was expressed as early as in aggregation streams, indicating that Dp87 become transcribed a few hours earlier than the other prespore-specific genes so far reported. This was confirmed by northern hybridization detected using an image plate analyzer. The fact that cells expressing Dp87 appeared at random in aggregation streams gives solid support to the idea that position-independent differentiation of prespore and prestalk cells, followed by their sorting, brings about pattern formation in this organism. PMID- 8404533 TI - The Drosophila Ras2 and Rop gene pair: a dual homology with a yeast Ras-like gene and a suppressor of its loss-of-function phenotype. AB - The promoter of the Drosophila melanogaster Ras2 gene is bidirectional, regulating an additional gene oriented in the opposite polarity. The two divergently transcribed genes are only 93 bases apart and deletion analysis proved that common cis-acting elements within this promoter region are required for the transcriptional activity of both genes. We cloned the gene paired with Ras2 in the bidirectional promoter and isolated cDNAs corresponding to its mRNA. The Ras opposite (Rop) gene encodes for a 68 x 10(3) M(r) protein which shares sequence homology with the members of a novel Saccharomyces cerevisiae gene family, including the SLY1, SEC1 and VPS33 (SLP1) genes, all of which are involved in vesicle trafficking among yeast cellular compartments. A highly conserved motif in this family is also found in beta-COP, a coat protein isolated from rat Golgi-bound nonclathrin vesicles. Thus, the Rop protein may be a component of one of the vesicle trafficking pathways in Drosophila cells. The Rop gene expression during embryogenesis is restricted to the central nervous system (CNS) and the garland cells, a small group of nephrocytes that takes up waste materials from the haemolymph by endocytosis. Ras2 is also expressed in the embryonic garland cells. In postembryonic stages, the two genes are co-expressed in the larval salivary glands and the central nervous system, and in the adult CNS and reproductive systems. Interestingly, the S. cerevisiae SLY1-20 allele is a suppressor of the loss of the YPT1 gene, a ras-like gene implicated in vesicle translocation, suggesting that the two genes may interact with one another. Since Sec1p and beta-COP may also interact with small GTP-binding proteins of the ras superfamily, it is conceivable that the Rop and Ras2 gene products are not just co-expressed in common tissues, but may also functionally interact with one another in these tissues. PMID- 8404534 TI - Chimeric analysis of T (Brachyury) gene function. AB - To investigate T(Brachyury) gene function, a chimeric analysis of midgestation (9.5-11.5 days post coitum) embryos has been performed. Embryonic stem (ES) cell lines homozygous or heterozygous for the T gene have been introduced into wild type host embryos by blastocyst injection, and the resulting chimeras scored for morphological abnormality and extent of colonization by T/T cells. As observed previously in earlier stage chimeras (Rashbass, P., Cooke, L. A., Herrmann, B. G. and Beddington, R. S. P. (1991) Nature 353, 348-350), 9.5-11.5 dpc T/T<==>+/+ chimeras exhibit many morphological features of intact T/T mutants. In addition, a dramatic bias of T/T cells towards caudal regions (such as tail and allantois) was observed in all chimeras tested. This is likely to result from accumulation of nascent T/T mesoderm cells with time near the primitive streak, possibly because of altered migration or adhesion properties. T/+ cells colonized rostral regions efficiently, but a slight bias towards the distal end of the tail was still evident. No such bias was observed in control chimeras. The presence of T/T cells in the allantois resulted in its failure to form a correct placental connection and thus arrested later development. In contrast, chimeras in which T/T cells were present predominantly in the tail developed normally but exhibited severe tail abnormalities such as foreshortening, branching and haemorrhagic cavities. Moreover, in these embryos, much higher levels of chimerism were present in the distal end of the tail than in younger (9.5 dpc) embryos. Later in gestation, such abnormal tails probably degenerated, giving rise to neonates with absent or severely abnormal tails but no evidence of chimerism. In situ analysis of T expression in the tail reveals that normally T is expressed highly in the tailbud (the growing portion of the tail) during its elongation between 9.5 and 11.5 dpc. Thus, evidence both from chimeras and from T expression in the tail suggest that T may play a role in the correct deployment of cells emerging from the tailbud. PMID- 8404535 TI - Isolation and characterisation of murine homologues of the Drosophila seven in absentia gene (sina). AB - The seven in absentia gene (sina) is required for formation of the R7 photoreceptor cell in the developing eye of Drosophila melanogaster. The sina protein contains a putative zinc finger domain and localises to the cell nucleus in Drosophila. We report here the identification of a family of genes in the mouse (designated Siah) with extensive sequence homology to Drosophila sina. The Siah genes fall into two main groups: Siah-1, which consists of four closely related members, two of which appear to be functional, and Siah-2, which contains a single functional member. The predicted Siah proteins show an unusually high degree of conservation with sina over the majority of their lengths, diverging significantly only at their amino terminal ends. The Siah-1 and Siah-2 genes are widely expressed at a low level in the embryo and adult. Analysis of Siah-2 by hybridisation histochemistry shows that it is expressed at a higher level in a restricted number of sites during development, including the olfactory epithelium, retina, forebrain and proliferating cartilage of developing bone. The striking degree of sequence homology observed between the Drosophila and murine genes implies strong conservation pressure on the Siah genes and suggests that they play a significant role in vertebrate development. PMID- 8404536 TI - Neurotrophin-4 is a target-derived neurotrophic factor for neurons of the trigeminal ganglion. AB - The cellular localization of mRNA for neurotrophin-4 (NT-4), a novel neurotrophic factor, in the developing whisker follicles and skin of the embryonic rat is demonstrated by in situ hybridization. Levels of NT-4 mRNA in the whisker pad decrease between embryonic day 13 (E13) and E20, correlating in time with the onset of naturally occurring neuronal death in the innervating trigeminal ganglion. In addition to NT-4, brain-derived neuotrophic factor (BDNF) mRNA is also shown to be expressed in the rat embryonic whisker follicles although in a different cellular localization, which combined with previous data on the expression of NGF and NT-3 mRNAs, shows that all four neurotrophins are expressed during development of this structure. NT-4 protein is shown to elicit neurite outgrowth from explanted embryonic trigeminal ganglia and to promote neuronal survival of dissociated trigeminal ganglion neurons when cultured during the phase of cell death. NT-4 and NT-3 mainly support different neuronal subpopulations, whereas some NT-4-responsive cells appear to respond also to NGF and BDNF. Analysis of mRNAs for members of the Trk family of neurotrophin receptors in neurons rescued by different neurotrophins demonstrates the presence of distinct neuronal subpopulations that respond to specific combinations of these factors. Based on these results we propose that NT-4, together with the other three neurotrophins, orchestrate the innervation of the different structures of the developing whisker pad by the trigeminal ganglion, acting as target-derived neurotrophic factors for different subpopulations of trigeminal ganglion neurons. PMID- 8404537 TI - Connexin trafficking and the control of gap junction assembly in mouse preimplantation embryos. AB - Gap junction assembly in the preimplantation mouse embryo is a temporally regulated event, beginning a few hours after the third cleavage during the morphogenetic event known as compaction. Recently, we demonstrated that both mRNA and protein corresponding to connexin43, a gap junction protein, accumulate through preimplantation development beginning at least as early as the 4-cell stage. Using an antibody raised against a synthetic C-terminal peptide of connexin43, this protein was shown to assemble into gap junction-like plaques beginning at compaction (G. Valdimarsson, P. A. De Sousa, E. C. Beyer, D. L. Paul and G. M. Kidder (1991). Molec. Reprod. Dev. 30, 18-26). The purpose of the present study was to follow the fate of nascent connexin43 during preimplantation development, from synthesis to plaque insertion, and to learn more about the control of gap junction assembly during compaction. Cell fractionation and reverse transcription-polymerase chain reaction were employed to show that connexin43 mRNA is in polyribosomes at the 4-cell stage, suggesting that synthesis of connexin43 begins at least one cell cycle in advance of when gap junctions first form. The fate of nascent connexin43 was then followed throughout preimplantation development by means of laser confocal microscopy, using two other peptide (C-terminal)-specific antibodies. As was reported previously, connexin43 could first be detected in gap junction-like plaques beginning in the 8-cell stage, at which time considerable intracellular immunoreactivity could be seen as well. Later, connexin43 becomes differentially distributed in the apposed plasma membranes of morulae and blastocysts: a zonular distribution predominates between outside blastomeres and trophectoderm cells whereas plaque-like localizations predominate between inside blastomeres and cells of the inner cell mass. The cytoplasmic immunoreactivity in morulae was deemed to be nascent connexin en route to the plasma membrane since it could be abolished by treatment with cycloheximide, and redistributed by treatment with monensin or brefeldin-A, known inhibitors of protein trafficking. Treatment of uncompacted 8-cell embryos with either monensin or brefeldin-A inhibited the appearance of gap junction-like structures and the onset of gap junctional coupling in a reversible manner. These data demonstrate that the regulated step in the onset of gap junction assembly during compaction is downstream of transcription and translation and involves mobilization of connexin43 through trafficking organelles to plasma membranes. PMID- 8404538 TI - Ventral spinal cord inhibition of neurite outgrowth from embryonic rat dorsal root ganglia. AB - Organotypic culture of embryonic rat lumbar spinal cord and dorsal root ganglia has been used to demonstrate an inhibitory effect of ventral spinal cord on neurite growth from dorsal root ganglion explants. When dorsal root ganglion explants from 14-15 day old embryos were cultured alone or in close proximity to a dorsal cord explant, the pattern of dorsal root ganglion neurite outgrowth was typically radial. However, when E14-15 dorsal root ganglion explants were cocultured for 22-24 hours in proximity to a ventral spinal cord explant from the same embryo, few, if any, dorsal root ganglion neurites grew in the direction of the ventral cord explant. This inhibitory effect appeared to be developmentally regulated; it was diminished or absent in cocultures prepared from 18 day old embryos. In contrast, in cocultures of dorsal cord and ventral cord explants from E14-15 embryos, dorsal cord neurites grew abundantly toward the ventral cord explant suggesting that the inhibition is not likely to be due to a nonspecific neurotoxic effect and that the activity responsible selectively inhibits dorsal root ganglion neurite outgrowth. We conclude that a diffusible, primary afferent inhibitory factor(s) produced by embryonic ventral horn may be responsible for the inhibition. Our results are discussed with respect to the possible involvement of inhibition in the normal development of primary afferent innervation of the spinal cord. PMID- 8404539 TI - Mechanisms of dorsal-ventral axis determination in Drosophila embryos revealed by cytoplasmic transplantations. AB - The establishment of the dorsal-ventral pattern in Drosophila embryos depends on a signal transduction process: a putative extracellular ligand released into the perivitelline space surrounding the embryo binds to the Toll receptor. Toll activation triggers the formation of the nuclear gradient of dorsal protein, the morphogen of the dorsal-ventral axis. Here, I analyse the dorsal protein distribution and the expression of zygotic dorsal-ventral genes in Toll- embryos that have been injected with wild-type cytoplasm under a variety of different injection conditions. Injections into two positions within a single embryo lead to the formation of two dorsal-ventral patterns in one embryo, allowing the analysis of interactions between pattern-forming processes. The results of single and double injections suggest that the spatial information for the embryonic dorsal-ventral axis is largely derived from spatial cues present in the extraembryonic compartment, which restrict the release of the putative Toll ligand. They argue against a Toll-dependent pattern-formation process employing local self-enhancement and lateral inhibition to enhance a weak initial asymmetry. The putative Toll ligand appears to originate from a ventrally restricted zone which extends along the entire anterior-posterior axis. Ligand diffusion or its graded release are required to determine the slope of the nuclear dorsal protein gradient. Both the Toll receptor and the putative ligand of Toll are in excess in wild-type embryos. Since spatial information for the embryonic dorsal-ventral axis is already present in the vitelline membrane or the perivitelline space, it is most likely generated during oogenesis. Oogenic pattern formation is also responsible for the perpendicular orientation the dorsal-ventral axis maintains with respect to the anterior-posterior axis. PMID- 8404540 TI - The cell adhesion molecule M-cadherin is specifically expressed in developing and regenerating, but not denervated skeletal muscle. AB - The spatiotemporal distribution of M-cadherin mRNA has been determined by in situ hybridization in the mouse embryo and in adult skeletal muscle following experimental regeneration and denervation. M-cadherin mRNA is highly tissue specific and is found only in developing skeletal muscle. In contrast, N-cadherin mRNA has a broader tissue distribution in the embryo, being found on both neural elements and skeletal and cardiac muscle. M-cadherin is expressed in the myotomes shortly after they form, along with the myogenic regulatory factor myogenin. M cadherin is expressed in muscles derived from the myotomes and is detected in forelimb bud precursor cells at embryonic day 11.5. In the latter case M-cadherin expression appears co-ordinately with that of myogenin and cardiac alpha-actin. Shortly before birth, M-cadherin expression is down regulated. M-cadherin can, however, be re-expressed following experimental regeneration of skeletal muscle. Here M-cadherin is transiently expressed on regenerating myoblasts but not myotubes. Following muscle denervation no evidence was found for re-expression of M-cadherin under conditions where there was strong expression of the nicotinic acetylcholine receptor on myofibres. The highly specific tissue distribution and unique developmental profile distinguishes M-cadherin from other cadherins and suggests a role in cell surface events during early myogenesis. PMID- 8404541 TI - Maintenance of ZPA signaling in cultured mouse limb bud cells. AB - The positional signal localized to the posterior (zone of polarizing activity or ZPA) region of the vertebrate limb is transiently expressed during development and a decline in ZPA signaling is accelerated when posterior cells are dissociated and cultured in vitro. The evidence that cultured posterior cells display a precocious decline in ZPA signaling when compared to in vivo studies suggests that a factor present in the limb bud maintains or stabilizes ZPA signaling during limb outgrowth and that this maintenance factor is lost and/or exhausted in in vitro studies. We have developed a new culture technique, 'microdissociation', which preserves extracellular components that we have found to be necessary for ZPA signal maintenance. Our data suggest that the limb bud ectoderm produces a maintenance activity that becomes stored in the extracellular matrix where it acts on limb bud cells to stabilize the activity of the ZPA signal. Using our initial characterization of this maintenance activity, we have identified a growth factor, FGF-2 (bFGF), that can replace all of the ZPA signaling maintenance activity observed in microdissociate cultures. The existence of various members of the FGF family in the developing limb strongly argues a role for FGF in stabilizing ZPA signaling in vivo. PMID- 8404542 TI - Contractile protein gene expression in primary myotubes of embryonic mouse hindlimb muscles. AB - The time course of contractile protein [actin, myosin heavy chain (MHC) and myosin light chain (MLC)] gene expression in the hindlimb muscles of the embryonic mouse (< 15 days gestation) has been correlated with the expression of genes for the myogenic regulatory factors, myogenin and MyoD, and with morphogenetic events. At 14 days gestation, secondary myotubes are not yet present in crural muscles (M. Ontell and K. Kozeka (1984) Am. J. Anat. 171, 133 148; M. Ontell, D. Bourke and D. Hughes (1988) Am. J. Anat. 181, 267-278); therefore, all transcripts for contractile proteins found in these muscles must be produced in primary myotubes. In situ hybridization, with 35S-labeled antisense cRNAs, demonstrates the versatility of primary myotubes in that transcripts for (1) alpha-cardiac and alpha-skeletal actin, (2) MHCembryonic, MHCperinatal and MHC beta/slow, and (3) MLC1A, MLC1F and MLC3F are detectable at 14 days gestation. While the general patterns of early activation of the cardiac genes and early activation of the genes for the developmental isoforms are preserved in both myotomal and limb muscles (D. Sassoon, I. Garner and M. Buckingham (1988) Development 104, 155-164 and G. E. Lyons, M. Ontell, R. Cox, D. Sassoon and M. Buckingham (1990) J. Cell Biol. 111, 1465-1476 for myotomal muscle), there are a number of differences in contractile protein gene expression. For example, in the myotome, when myosin light chain genes are initially transcribed, hybridization signal with probe for MLC1A mRNA is greater than that with probe for MLC1F transcripts, whereas the relative intensity of signal with these same probes is reversed in the hindlimb. The order in which myosin heavy chain genes are activated is also different, with MHCembryonic and MHCperinatal preceding the appearance of MHC beta/slow transcripts in limb muscles, while MHCembryonic and MHC beta/slow appear simultaneously in the myotomes prior to MHCperinatal. In the myotome, an intense hybridization signal for alpha-cardiac and a weak signal for alpha-skeletal actin transcripts are detectable prior to myosin mRNAs, whereas in the limb alpha-cardiac actin transcripts accumulate with myosin transcripts before alpha-skeletal actin mRNA is detectable. These differences indicate that there is no single coordinate pattern of expression of contractile protein genes during initial formation of the muscles of the mouse.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404543 TI - Defects of embryonic organogenesis resulting from targeted disruption of the N myc gene in the mouse. AB - The highest expression of the N-myc gene occurs during embryonic organogenesis in the mouse ontogeny, with the peak of expression around embryonic day 9.5. Homozygous N-myc-deficient mice, produced by germline transmission of a disrupted allele in ES cells, developed normally to day 10.5, indicating dispensability of N-myc expression in the earlier period, but later accumulated organogenic abnormalities and died around day 11.5. The most notable abnormalities were found in the limb bud, visceral organs (lung, stomach, liver and heart) and the central/peripheral nervous systems, and were highly correlated with the site of N myc expression. The limb buds and the lungs excised from N-myc-deficient mutant embryos were placed in culture to allow their development to stages beyond the point of death of the embryos. Analyses indicated that the mutant limbs failed to develop distal structures and the development of bronchi from the trachea was defective in the lungs. The latter defect was largely corrected by addition of fetal calf serum to the culture medium, suggesting that an activity missing in the mutant lung was replenished by a component of the serum. The phenotype of N myc-deficient mutant embryos indicated requirement of the N-myc function in many instances of tissue interactions in organogenesis and also in cell-autonomous regulation of tissue maturation. PMID- 8404544 TI - Cytotoxicity of tacrine and velnacrine metabolites in cultured rat, dog and human hepatocytes. AB - Clinical trials with tacrine (THA) and its principal (1-OH) metabolite (velnacrine) for the treatment of Alzheimer's disease have been hampered by adverse hepatic events that were undetected in preclinical studies. As part of integrated in vivo/in vitro efforts to characterize the role of metabolites in these events, cultured cells were evaluated for their suitability for further mechanistic studies. The relative cytotoxic potentials of THA, three monohydroxy metabolites of THA (including velnacrine, a racemate), the two velnacrine enantiomers, and several known and suspected dihydroxy velnacrine metabolites were determined. Cytotoxicity was evaluated in 24-hour cultures by morphology and by the Neutral Red Uptake Assay. All test articles were evaluated in primary rat hepatocytes and in a human hepatoma cell line (HepG2). THA and velnacrine were also tested in a rat hepatoma cell line (H4) and in primary dog hepatocytes. The metabolic competency of each cell type was determined. Sensitivity to THA and velnacrine was greatest in H4 cells, followed by primary rat and HepG2 cells; dog cells were least sensitive. In HepG2 cells, THA was clearly more cytotoxic (LC50:54 micrograms/ml) than its monohydroxy metabolites (LC50 values: 84 to 190 micrograms/ml); dihydroxy velnacrine metabolites were the least cytotoxic (LC50 values:251 to 434 micrograms/ml); the relative order was comparable in primary rat hepatocytes. Roles for reactive metabolites and/or altered metabolic capabilities of Alzheimer's patients are suggested. PMID- 8404545 TI - Effect of sodium selenite on morphine-induced hepatotoxicity in mice. AB - The effects of selenite on morphine hepatotoxicity and metabolism were examined. Pretreatment of male mice with sodium selenite (2 mg/kg, ip) 24 hr before morphine administration significantly protected them against the hepatotoxic effects of morphine as evidenced by decreased plasma alanine and aspartate transaminase values. This treatment inhibited the decrease in hepatic glutathione content that occurs in animals receiving hepatotoxic doses of morphine. Selenium pretreatment decreased the in vivo covalent bonding of morphine to hepatic proteins. Selenium also lowered the in vitro covalent bonding of morphine to hepatic microsomal proteins. PMID- 8404546 TI - Subacute immunotoxicity study of formaldehyde in male rats. AB - Immune functions were examined in male rats following 28 day oral administration of formaldehyde by gastric tube at dose levels of 0, 20, 40, and 80 mg/kg. Routine parameters examined included hematology, clinical chemistry, and body, thymus, kidney, and liver weights. In addition, cellularity of spleen and lymph nodes, histology of spleen, thymus, lymph nodes, liver kidney, small and large intestines, and histochemistry of spleen and lymph nodes were evaluated. Immune parameters evaluated included serum hemagglutinin antibody response; antibody plaque forming cell response to sheep erythrocytes (lymphocyte-dependent antigen); microbicidal activity of Candida albicans; and phagocytic activity by adhesion of microspheric hydrophilic synthetic particles to leukocyte cell membrane. Body weights were slightly decreased at high dose level (80 mg/kg). The difference was significant when compared to the controls. The lymph node weights were significantly increased in rats receiving formaldehyde. The cellularity of lymphoid organs was not influenced after 28 day exposure to formaldehyde. PMID- 8404547 TI - Ten and ninety-day toxicity studies of 2-chlorophenol in Sprague-Dawley rats. AB - Male and female Sprague-Dawley rats received 2-chlorophenol in corn oil daily by gavage for 10 or 90 consecutive days. The 10-day study doses were 13, 64, 129 and 257 mg/kg while the 90-day subchronic study doses were 17, 50 and 150 mg/kg. In the 10-day study, hematologic and clinical chemistry, food and water consumption, absolute and relative organ weights, and histopathological findings revealed no compound or sex-related effects. In the 90-day study there were no significant gross or histopathological findings that were treatment-related in either sex. There were statistically significant differences between control and treated groups associated with hematology, clinical chemistry and organ weights; however, none of the differences were considered to be biologically meaningful. PMID- 8404548 TI - Subchronic toxicity study of 1,1-dichloro-2-propanone in Sprague-Dawley rats. AB - Groups of 10 male and 10 female Sprague-Dawley rats were administered 1,1 dichloro-2-propanone in corn oil by gavage at 0, 10, 20, 40, or 80 mg/kg/day for 90 consecutive days. Food and water consumption, body and organ weights, organ-to body weight ratios, hematology, and clinical chemistry parameters were determined. Gross and microscopic pathology examinations also were conducted. No treatment-related mortality was observed during the study; however, liver, forestomach, and kidney toxicity was evident. Liver changes consisted of cytoplasmic alteration, cytomegaly, karyomegaly, and bile duct hyperplasia. These occurred with significance of p < or = 0.05 at or above 10 mg/kg/day in both sexes. The forestomach lesions included hyperkeratosis and epithelial hyperplasia in both sexes at 40 and 80 mg/kg/day, and ulcerations at 80 mg/kg/day. Also, an increased incidence and severity of spontaneously occurring chronic progressive nephropathy was most apparent in high dose males. Increases in organ-to-body weight ratios were noted for the liver and kidneys in females at the highest dose level and in males at the two highest dose levels. Serum enzymes (ALT, AST, and LDH) were increased in females and decreased in males. Based on liver lesions and biochemical changes, it was concluded that there was no experimentally definable NOAEL. PMID- 8404549 TI - Neural dopamine D2 receptors in rats fed endophyte-infected fescue seed. AB - To study the effect of endophyte (Acremonium coenophialum) on hypothalamic and striatal dopamine D2 receptors, male rats (n = 14/group) were pair-fed diets containing 50% Rat Chow and 50% either endophyte-infected (E+) or noninfected (E ) fescue (Festuca arundinacea Schreb.) seed for 21 days. Concentrations of ergovaline and saturated pyrrolizidines were 1.91 micrograms/g and 2.84 mg/g, respectively in E+, and undetectable in E- fescue seed. To monitor endophyte effects, rats were weighed weekly and serum derived from trunk blood (d 21) was analyzed for prolactin. Corpus striatum and hypothalamic tissue was assayed for dopamine D2 receptors using [3H]spiperone and [125I]epidepride, respectively. The endophyte depressed (P < .06) serum prolactin concentrations. Average daily gain during the study (21 d) was depressed (P < .0043) in rats fed E+ compared to controls. The endophyte increased (P < .03) striatal D2 receptor affinity (KD = 48.70 vs 54.95 pM) with no change (P > .28) in receptor density (Bmax = 25.59 vs 28.00 pmol/mg of tissue) in E+ and E- rats, respectively. Hypothalamic D2 receptor density (Bmax = 1.79 vs 1.57 pmol/mg of tissue) and affinity (KD = 17.5 vs 17.26 pM) were not (P > .66) different between E+ and E- rats, respectively. These data suggest changes in D2 receptor binding characteristics, particularly receptor affinity, may contribute to signs of fescue toxicosis. PMID- 8404550 TI - Long-term ventilation myringostomy. PMID- 8404551 TI - Vocal fold scar and vocal fold nodules. PMID- 8404552 TI - Endoscopic view of maxillary sinus ostia. PMID- 8404553 TI - Electrocautery versus epinephrine-injection tonsillectomy. AB - This prospective, single-blinded study on tonsillectomy was done to compare the Bovie electrocautery dissection technique with the epinephrine-and-lidocaine injection technique and to document which technique is safer, faster, and less morbid. Twenty-nine patients who were scheduled for tonsillectomy at two Northern California Region Kaiser Permanente Medical Centers were enrolled in the study. Each patient served as his or her own control, and tonsillectomy technique was randomized on the basis of even or odd numbers of the last digit of the medical record number. The time of injection of epinephrine and time of dissection for each tonsil was recorded. Blood loss was quantified for each dissection, and pain was assessed by asking the patient which side hurt more. I found no statistically significant difference in operating time, intraoperative blood loss, or postoperative hemorrhage between these two methods. The electrocautery, or Bovie, technique produced more clinically significant eschar and delayed healing than the epinephrine-injection technique did. The Bovie technique produced more inadvertent burns to surrounding tissue than the epinephrine-injection technique did, but the epinephrine caused transient tachycardia in 14% of the study participants. The present study showed no difference between the two techniques in postoperative pain experienced by study participants, but other recent studies have shown that patients experience more pain when the Bovie technique is used. Study data do not support the superiority of Bovie tonsillectomy. I recommend epinephrine-injection tonsillectomy as a safe and expedient operation. PMID- 8404554 TI - Tonsil and adenoid surgery for airway obstruction: perioperative respiratory morbidity. AB - A series of 160 consecutive patients undergoing tonsil and adenoid surgery for upper airway obstruction is reported. The ages ranged from 8 months to 13 years. Sixty-seven percent were 2, 3, or 4 years of age. All were routinely admitted overnight postoperatively. Forty-five (28%) remained in the hospital longer than one night (2 to 20 days). Postoperative respiratory problems were the reason for prolonged hospital stay in 30 of these 45 patients. Preoperative "danger-signals" of potential postoperative respiratory problems were: a history of severe obstructive symptoms with apnea and moderate or strongly positive sleep study, daytime somnolence, need for urgent T&A, and cardiomegaly. Risk factors present in a smaller number of patients were obesity, congenital stenosis of airways, and bronchopulmonary dysplasia. We suggest that children with these danger signals not be considered as candidates for outpatient T&A surgery. PMID- 8404555 TI - Fungal osteomyelitis of the temporal bone: a review of reported cases. PMID- 8404556 TI - Laryngeal reinnervation in children: the Leeds experience. PMID- 8404557 TI - Endoscopic sinus surgery: sinonasal polyposis and allergy. PMID- 8404558 TI - Control of early postoperative pain with bupivacaine in pediatric tonsillectomy. AB - Early postoperative pain following pediatric tonsillectomy remains a significant obstacle to speedy recovery and smooth convalescence. Inadequate analgesia causes poor oral intake and occasionally requires overnight hospitalization in same day surgery practices. Several otolaryngologists anecdotally support intraoperative infiltration with a long-acting amide anesthetic, bupivacaine hydrochloride, for postoperative pain control. However, no previous study supports these claims. At the National Naval Medical Center, a prospective, randomized, double-blinded study was undertaken in fifty patients, ages 3 to 16 years old. After tonsil resection, either 0.5% bupivacaine or saline was injected into each fossa. We asked the parents three questions. First, what was the level of pain at 2, 6 and 10 hours postoperatively? Second, what was the amount of oral intake during the first 10 hours after surgery? And third, how many doses of oral acetaminophen elixir were administered over the first 10 postoperative hours? Bupivacaine administration caused no adverse effects. There was no difference in perceived pain level between bupivacaine and saline groups at 2, 6 and 10 hours. Oral intake levels over the first 10 hours were similar. Although bupivacaine group patients received fewer doses of oral acetaminophen, the difference between groups was not statistically significant. We conclude that bupivacaine is a safe medication for infiltration, but offers no advantage over saline in the control of early postoperative pain in pediatric tonsillectomy. PMID- 8404559 TI - The pediatric sinusitis war. PMID- 8404560 TI - Effects of cutaneous stimulation on ipsilateral and contralateral motoneuron excitability: an analysis using H reflexes and F waves. AB - F waves are altered by conditioning stimulation of digital nerves. We investigated this by studying series of median nerve F waves recorded from abductor pollicis brevis (APB) 20, 50, 80, 100, 150, 200, and 500 milliseconds (ms) after conditioning stimuli applied to the ipsilateral or contralateral second digit in six normal subjects. The persistence and mean amplitude of the F waves were determined at each delay and compared with control F waves without conditioning stimuli. Ipsilateral afferent stimulation produced inhibition of F wave amplitude and persistence when applied 50 to 100 ms before the test stimulus, while contralateral conditioning stimuli facilitated the F wave when applied 80 to 200 ms before the test stimulus. Similar results were obtained in H reflex studies in one neurologically normal subject with a reproducible H reflex recording from the APB. These findings illustrate definable ipsilateral and contralateral postsynaptic influences of afferent fibers on motoneuron excitability in the arms. PMID- 8404561 TI - Magnetically elicited blink reflex: an alternative to conventional electrical stimulation. AB - Magnetically and electrically elicited blink reflexes were studied in 11 normal subjects and in 5 patients with partial peripheral facial nerve palsy. Both methods were able to elicit a reproducible blink reflex. In both groups the latencies of the magnetically elicited reflexes were equal to those that followed electrical stimulation. The magnetic stimulation technique offered 2 advantages over the electrical stimulation: (1) it was less painful and therefore better tolerated and (2) the coil position in the middle of the forehead was sufficient to elicit a bilateral blink reflex because the windings of the stimulation coil excited both supraorbital nerves simultaneously. This bilateral response enables one to reduce the number of stimuli and can be used in the examination of patients that have peripheral facial nerve palsies without the concomitant impairment of the reflex pathway in the brainstem. The magnetic technique can also elicit a blink reflex at stimulation points distant from the supraorbital notch. This finding offers an alternative explanation of the late responses of facial muscles following transcranial magnetic stimulation of the motor cortex. PMID- 8404562 TI - Inhibition of EMG activity in isometrically loaded agonist muscle preceding a rapid contraction. AB - It was recently suggested that a premotion silent period in isometrically loaded agonist muscle reflects transition process in switching motor program from an isometric to an isotonic condition. To test this hypothesis we investigated changes in EMG activity of a biceps muscle during motor preparatory phase in an entirely isometric paradigm. Five healthy volunteers produced rapid isometric elbow flexions superimposed on a slightly sustained contraction during self-paced and reaction time conditions. In addition to surface EMG, single motor units were recorded in agonist muscle prior to a rapid contraction. Silent period was found in responses of three out of five subjects, whereas inhibition of some but not all tonically active motor units was observed in all studied subjects prior to the phasic EMG burst. Inhibition of motor unit activity occurred more often in self-paced than reaction time contractions. Our results indicate that inhibition is not always powerful enough to produce a complete electrical silence, and weaker inhibitory effects were observed as a declining number of firing motor units. We conclude that the silent period or depression of the EMG activity is associated with changes in motor program from tonic to phasic muscle activation and is not merely a consequence of changes from an isometric to an isotonic condition. PMID- 8404563 TI - Evoked potentials in workers occupationally exposed to organic solvents. AB - Somatosensory, Brainstem Auditory and Pattern Reversal Evoked Potentials (SEPs, BAEPs, PREPs) are recorded in workers occupationally exposed to mixtures of organic solvents, in order to specify the levels of the nervous system affected by a long term exposure to solvents, and to analyze the effects of age and gender. The most significant differences are found for SEPs; they objectivate peripheral impairments magnifying the differential effects of age observed in the control subjects, and show the higher sensitivity of women. The central impairment is pointed out by the latency delay of P22 component mainly, whose age related increase is amplified by solvent exposure. PMID- 8404564 TI - Medial antebrachial cutaneous nerve conduction in true neurogenic thoracic outlet syndrome. AB - Thoracic outlet syndrome (TOS) and its diagnosis have been topics of controversy, fueled in part by the lack of an objective test for early diagnosis. True neurogenic TOS results in diagnostic EMG findings only in its later stages. We describe a series of patients in whom the diagnosis is supported by changes in the medial antebrachial cutaneous (MAC) nerve at or before the development of these typical EMG findings in median and ulnar nerves. PMID- 8404565 TI - Antidromic sensory nerve conduction studies of medial and lateral plantar nerves in normals. AB - A reliable sensory nerve conduction study for the most distal lower extremities is needed in routine clinical electromyography. This paper reports a study of 150 medial and lateral plantar nerves in the foot in normals. An antidromic technique was used with stimulation at the ankle and recording from the small and large toes. Recordable responses were obtained in 149 instances. Conduction velocity in the medial branch distal to the tarsal tunnel was 40.5 +/- 4.0 m/sec and significantly slower than conduction in the lateral branch by 4.3 m/sec. The amplitude of the evoked response from the big toe was 3.46 +/- 2.2 microV and significantly larger than that in the small toe by 1.34 microV. Evoked response amplitude in the medial branch was greater in younger subjects. PMID- 8404566 TI - Duration, configuration and amplitude of the motor response evoked by magnetic brain stimulation in patients with multiple sclerosis. AB - Analysis of motor evoked potentials (MEP) after magnetic brain stimulation is often restricted to measurement of latency and amplitude. In the present study, which was conducted in healthy controls and in patients with multiple sclerosis (MS), the additional value of MEP duration and configuration was evaluated. Recordings were made from distal muscles of the upper and lower extremity. In the MS group, mean MEP duration was found to be prolonged, while amplitudes were reduced. MEP of abnormally prolonged duration, as compared to normative data derived from the control group, were found even in presence of normal central motor conduction times. These findings are compatible with increased temporal dispersion of the impulses arriving at the spinal motoneurone pool. Thus, analysis of MEP duration may provide additional information about impulse propagation along the descending motor tracts, indicating disturbed conduction properties even in the presence of normal central motor conduction time. In contrast, MEP configuration was of no value for differentiating between the two groups. PMID- 8404567 TI - Further evidence of functional differentiation within biceps brachii. AB - This study investigated whether the two heads of Biceps Brachii could be functionally differentiated during rapid supination movements with different degrees of elbow flexion, shoulder axial rotation and load. Surface electromyograms, recorded from the long and short heads of Biceps Brachii, were utilised to identify changes in the intensity of muscle contraction. Based upon an analysis of sixteen subjects, the results indicated that joint position (muscle length), but not load, significantly (p < 0.05) influenced the relationship between the contraction intensities of the two heads. Specifically, increasing the length of the Biceps Brachii promoted increased activation of the long head in relation to the short head in producing rapid supination motions. It was concluded that functional differentiation within the two heads of Biceps Brachii was present in motions involving its distal insertion. PMID- 8404568 TI - Median nerve electrophysiologic parameters and psychomotor performance in carpal tunnel syndrome. AB - Psychomotor performance (PMP) involving a repeated, rapid pinch and release task was correlated with median nerve electrophysiologic parameters for control subjects (16 hands) and subjects with carpal tunnel syndrome (CTS) (14 hands). The psychomotor task was used because of its functional resemblance to many work related activities. A strain gauge dynamometer was repeatedly pinched to a predetermined force level using the index finger and thumb and then released as rapidly as possible, while measuring the actual isometric force exerted. Discrete visual and auditory feedback was provided. Median and ulnar nerve motor latencies and amplitudes, as well as median antidromic sensory latencies and amplitudes, and transcarpal latencies and amplitudes were obtained. CTS subjects had longer median motor and sensory latencies and were weaker than controls, however median motor and sensory amplitudes were not statistically different. A strong relationship was observed between electrophysiologic variables and PMP, which could not be accounted for by age differences alone. It is unclear whether the measured differences in PMP are related to sensory or motor deficits. PMID- 8404569 TI - Historical perspectives on law, medical malpractice, and the concept of negligence. AB - Since the dawn of human history, the inherent morbidity and mortality of human beings has made the diagnosis and treatment of human disease a high-risk profession. The ancient risk of physical retribution against the physician has been replaced by the modern risk of economic indemnity or compensation. Monetary settlements and awards are at times so huge that they may result in personal bankruptcy and professional disgrace. This article should not discourage health care providers from continuing their pursuits, but encourage them to enhance their knowledge about how and why medical malpractice has developed. It has been intended to facilitate the "prudent practitioner" with a more thorough understanding of some of the elements of negligence that have caused concerns in the past and will certainly create new concerns as science continues to drag the law in its wake. PMID- 8404570 TI - Consent and refusal of treatment. AB - Clearly, a multitude of potential consent problems can exist for the emergency physician. It is difficult at times to balance the concepts of patient autonomy with the desire to provide optimal medical care. Experienced emergency physicians should be able to individualize the type of consent needed in a particular situation, based on a clinical evaluation of the case. Some general principles apply, but no strict rules can guide the physician in every case. Documentation of consent and refusal of treatment are critical for quality of patient care and legal liability reasons. Principles of what is good, "appropriate" legal consent usually follow from good medical care and strict concern for the patient's health and rights. PMID- 8404571 TI - Minors and emergency medicine. AB - This article reviews medical-legal issues with regard to minors as they apply to the practice of emergency medicine. The topics of consent to treatment, refusal of care, child abuse, and medical malpractice are addressed. A working knowledge of these areas helps the emergency physician to avoid confusion, avoid delays in management, and maintain the safety of children. PMID- 8404572 TI - Legal considerations in prehospital care. AB - This article presents an overview of several major legal issues in contemporary prehospital health care delivery and emergency medical services. It includes review and analysis of medical-legal issues in medical control; patient consent and treatment; modern emergency medical services communications; discussion of medical-legal concerns in regard to patient destination choice, diversion, and transfer; and an analysis of theories of recovery including a review of selected case histories. PMID- 8404573 TI - Emergency medicine, psychiatry, and the law. AB - The emergency department physician must assess, initiate treatment, and arrange for the disposition of patients whose presentation reflects disordered thinking, emotions, and behavior. Unique diagnostic, management, and medicolegal challenges must be met daily. Understanding the issues inherent in the various stages of involvement with such patients helps reduce medicolegal risk and facilitates the provision of appropriate health care. PMID- 8404574 TI - The emergency department medical record. AB - Emergency physicians are overwhelmed by the load of medical record documentation in the emergency department. This article reviews the important reasons for better documentation and today's increasing regulatory requirements in medical record documentation. PMID- 8404575 TI - Risk management and high-risk issues in emergency medicine. AB - Risk management in the emergency department is defined in this article. The health care professional should focus on the health and best interests of the patient. PMID- 8404576 TI - The National Practitioner Data Bank. An overview for the emergency physician. AB - This article reviews the Health Care Quality Improvement Act, Title IV of Public Law 99-660, which was passed by the US Congress in 1986. The Act offered immunity to professional review societies of hospital and other health care entities, and persons serving on or otherwise assisting such bodies. It is believed that professional review actions will further the quality of medical care and provide due process safeguards for the health care provider. PMID- 8404578 TI - Leaders and leadership. PMID- 8404577 TI - Future legal issues in emergency medicine. AB - This article analyzes past legal trends in emergency medicine with an effort to project those trends and current developments into future legal issues that will confront emergency physicians and emergency departments. Special emphasis is placed on insurance trends and professional liability insurance developments along with medical malpractice claims past, present, and future. Also discussed in this article is the health care industry environment and ways that it might affect future legal challenges for emergency medicine. PMID- 8404579 TI - Leadership and vision: the library's role. PMID- 8404580 TI - Leadership and vision. PMID- 8404581 TI - Leadership changes and challenges. PMID- 8404582 TI - Leadership in research: Harbor-UCLA Medical Center experiences. PMID- 8404584 TI - Thoughts on organizational change. PMID- 8404583 TI - The doctor, the bus and the hat. PMID- 8404585 TI - Leadership. PMID- 8404586 TI - The spirit of nurse empowerment: a leader's responsibility. PMID- 8404587 TI - Thyrotropin--to N-link or unlink? That is the question. PMID- 8404588 TI - Purification and characterization of recombinant human thyrotropin (TSH) isoforms produced by Chinese hamster ovary cells: the role of sialylation and sulfation in TSH bioactivity. AB - The biological significance of glycosylation variants of pituitary glycoprotein hormones remains controversial because of the indirect methods usually employed to determine carbohydrate composition or structure as well as the use of unreliable biological/immunological ratio to determine bioactivity. We have previously characterized recombinant human TSH (rhTSH) secreted by Chinese hamster ovary cells attached to microcarrier beads in a large scale bioreactor after stable transfection of hCG alpha and hTSH beta minigenes. In the present study rhTSH has been used as a model to determine structure-function relationships of different isoforms of glycoprotein hormones. We have now produced greater than 200 mg rhTSH using a hollow fiber bioreactor. The highly purified rhTSH produced in the hollow fiber bioreactor (rhTSH-N) as well as rhTSH commercially produced in a large scale bioreactor (rhTSH-G) were quantitated by immunoassays, receptor binding assay, and amino acid analysis and further characterized by a variety of physico-biochemical methods, including chromatofocusing and carbohydrate analysis. rhTSH-G, rhTSH-N, as well as pituitary human TSH (phTSH) have been separated by chromatofocusing on a Mono P column into several isoforms with different pI values. Compositional analysis of the fractions showed higher sialic acid content in the more acidic rhTSH-G fractions. phTSH acidic isoforms showed higher total sulfate and sialic acid contents than the more basic fractions. The bioactivities of various TSH isoforms based on rigorous quantitation of mass by amino acid analysis determined in three different FRTL-5 cell bioassays showed that the more basic and less sialylated fractions of rhTSH-G were more active than the more acidic fractions. In contrast to the in vitro data, highly sialylated and acidic rhTSH-G isoforms showed longer plasma half-lives and higher in vivo bioactivity than the basic forms. These results indicate that secreted rhTSH, similar to intrapituitary phTSH, exists as a mixture of charge isoforms that are related at least in part to the degree of sialylation. The degree of sialylation, highly dependent on the bioreactor production conditions, appears to be the major factor affecting the charge heterogeneity, MCR, and bioactivity of rhTSH. PMID- 8404589 TI - Up-regulation of oxytocin receptors in rabbit amnion by glucocorticoids: potentiation by cyclic adenosine 3',5'-monophosphate. AB - Oxytocin (OT) receptors (OTR) in rabbit amnion increase more than 200-fold at the end of gestation. In the present report, we studied the basis of this up regulation. Incubation of amnion cells with cortisol (20 nM) for 24 h increased the amount of 125I-labeled OT antagonist bound by 16- to 18-fold. The effects of cortisol were dose and steroid dependent. Administration of glucocorticoid to pregnant does also increased the concentration of OTRs in amnion. The effects of cortisol in vitro were potentiated by the addition of forskolin (50 microM), so that OTR number increased by as much as 182 times. The effects of cortisol and forskolin, either separately or in combination, were inhibited by activation of protein kinase-C or coincubation with transforming growth factor-alpha (10 nM). Cyclosporin-A (5 microM) selectively inhibited cortisol-induced rises in the OTR concentration. The addition of cortisol to amnion cells increased OT-stimulated prostaglandin E2 (PGE2) release almost 100-fold; the combination of forskolin and cortisol increased the PGE2 response to OT about 5600 times. Judging from the greater effects on PGE2 release, these results suggest that forskolin and cortisol up-regulate the signal response mechanism to OT as well as the OTR concentration. The findings show that changes occurring in the amnion in vivo can be mimicked in vitro, and they elucidate the mechanism of up-regulation of OTR concentrations. PMID- 8404590 TI - Ambient light modifies gonadotropin-releasing hormone pulse generator frequency in the rhesus monkey. AB - In the course of previous studies using continuous monitoring of the electrophysiological correlates of GnRH pulse generator activity, characterized by episodic increases in hypothalamic multiunit electrical activity (MUA volley), it was noticed that the nocturnal slowing of pulse generator frequency was an acute phenomenon observable in the first MUA volleys after the lights were turned off, as was the increase in frequency when the lights were turned on in the morning. This suggested that the reduction in pulse generator frequency at night may not be the consequence of an intrinsic diurnal rhythm, but an effect of light per se. Indeed, as reported herein, such an effect was observed when the lights were turned on or off at times other than the normal illumination period (normal light schedule, lights on from 0700-1900 h). That this was not simply a response to arousal was shown by awakening the animals with loud recorded noises in total darkness at the same unaccustomed times without a resulting change in frequency. This suggests that the effect of light is specific, perhaps mediated by the retino-hypothalamic tract. This direct action of light, however, is superimposed upon a diurnal rhythm, as shown by a reduction in pulse generator frequency during the subjective night when the monkeys were kept in constant light or constant darkness. PMID- 8404591 TI - Ovarian thecal/interstitial androgen synthesis is enhanced by a follicle stimulating hormone-stimulated paracrine mechanism. AB - To obtain direct evidence for FSH-stimulated paracrine signaling in the ovary, 21 day-old intact or hypophysectomized female Wistar rats received four sc injections of recombinant human FSH (rhFSH; total dose, 16-72 IU) at 12-h intervals. Ovaries were removed 48 h after the first injection to extract total RNA for Northern analysis of 17-hydroxylase/C17-20-lyase (cytochrome P450c17 alpha) mRNA or to isolate thecal/interstitial cells for assessment of basal and hLH-responsive androgen synthesis in vitro. In situ hybridization with a 35S labeled cytochrome P450c17 alpha cRNA probe confirmed that expression of the cytochrome P450c17 alpha gene was specific to thecal/interstitial cells. The approximately 2.0-kilobase P450c17 alpha mRNA signal in ovarian total RNA from intact animals was increased approximately 5-fold by treatment with rhFSH (total dose, 72 IU) or PMSG (15 IU). This effect was shown to be dose dependent, with a approximately 2-fold increase in response to 16 IU (total dose) rhFSH. P450c17 alpha mRNA levels in isolated granulosa and thecal/interstitial cell total RNA from intact animals were compared to establish which was the principal cellular site of P450c17 alpha mRNA expression. The P450c17 alpha mRNA signal was undetectable in control granulosa cells and only barely discernible after treatment with 72 IU (total dose) rhFSH. In contrast, P450c17 alpha mRNA was abundant in control thecal/interstitial mRNA, and its level was increased 4- to 6 fold by treatment with rhFSH. Treatment of hypophysectomized animals with rhFSH did not consistently alter ovarian P450c17 alpha mRNA levels. During culture for 48 h in serum-free medium, basal androgen (androstenedione plus androsterone) production by thecal/interstitial cells from intact animals was unaffected by treatment with rhFSH in vivo, but hLH-stimulated androgen production by these cells was enhanced approximately 2-fold. Neither basal nor hLH-responsive androgen production by thecal/interstitial cells from hypophysectomized animals was altered by previous treatment with rhFSH in vivo. Treatment of thecal/interstitial cell cultures from both intact and hypophysectomized animals with inhibin (0.1-30 ng/ml), a putative granulosa-derived paracrine factor, did not measurably affect basal androgen synthesis, but potently enhanced LH responsive androgen synthesis in vitro. Similarly, treatment of thecal/interstitial cell cultures with conditioned medium from FSH-treated granulosa cell cultures significantly enhanced LH-responsive, but not basal, androgen production. We conclude that treatment of pituitary-intact rats with "pure" FSH modulates thecal/interstitial cell androgen synthesis. Granulosa cells, but not thecal cells, possess FSH receptors, and thecal/interstitial cells are the principal ovarian sites of P450c17 alpha expression.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404592 TI - Degradation of insulin by isolated rat renal cortical endosomes. AB - It has been widely accepted that in kidney, degradation of insulin occurs in lysosomes. It is thought that after internalization into the cell, insulin dissociates from its receptor, which then recycles to the plasma membrane, while the hormone is transported in endosomes to the lysosomes, where it is degraded. However, earlier studies from this laboratory have suggested that insulin may also be degraded in an extralysosomal site, most likely endosomes. Indeed, studies in other tissues, most notably liver, have shown that insulin degradation does take place in endosomes. Since the intracellular processing of insulin differs between different tissues and cell types, and as the kidney is a major site of insulin degradation, we set out to determine directly whether endosomes degrade internalized insulin in the kidney. Rats were injected with [125I]monoiodoinsulin, labeled at either the A14 or B26 tyrosine. After killing, the kidney cortex was excised, and heavy endosomes were prepared by differential and isopycnic centrifugation. The isolated [125I]insulin-loaded endosomes were incubated for up to 60 min in intracellular medium, and degradation of [125I] insulin was estimated by means of precipitation in trichloroacetic acid. In the presence of ATP (10 mM), the percent degraded was increased over the control value (no ATP present), but under these circumstances, degradation was greater when the endosomes contained internalized 125I-labeled [B26]insulin than with A14 labeled [125I]insulin (26% vs. 13% degraded/h). In the absence of ATP, the percent degraded increased when the pH of the incubation medium was lowered. Radiolabeled material was extracted from endosomes, and Sephadex G-50 analysis revealed the presence of high mol wt, insulin-size, and low mol wt material. Reverse phase HPLC analysis of the insulin-size material revealed the presence of intact insulin and a number of degradation products. The elution profiles of some of these products were consistent with that reported to arise from the action of the insulin-degrading enzyme. Western blot analysis with the antiinsulin degrading enzyme monoclonal antibody 9B12 confirmed the presence of the enzyme in endosomal preparations. We conclude that degradation of insulin does occur in kidney cortical endosomes, probably involves the insulin-degrading enzyme, and results in the formation of relatively large intermediate products as well as low mol wt products. PMID- 8404593 TI - Induction of the estrogen receptor by growth hormone and glucocorticoid substitution in primary cultures of rat hepatocytes. AB - Hepatic estrogen receptors (ER) mediate estrogenic effects on mammalian liver metabolism and are thereby involved in the regulation of important physiological/pathological processes, such as coagulation, atherosclerosis, and hypertension. The regulation of the formation of the ER in primary cultures of rat hepatocytes was studied by assaying ER and ER mRNA under different endocrine conditions. The ER concentration was measured using two different methods, a ligand-binding technique and an ER enzyme immunoassay. The results obtained by the two methods showed good correlation, and linear regression analysis gave a correlation coefficient of 0.95. ER concentrations fell to low steady state levels within 16 h after establishing the cell culture and remained low in the absence of hormonal substitution. Upon medium supplementation with pituitary GH and the glucocorticoid dexamethasone (DEX) in combination, the ER concentration increased 6-fold from 4.2 +/- 1.0 to 25.8 +/- 7.0 fmol/mg cytosolic protein. ER mRNA was measured by solution hybridization. Substitution with GH and DEX in combination increased ER mRNA to 210 +/- 14% of control levels. No effect on ER mRNA stability was seen after hormone treatment. It is concluded that the regulatory effects of GH and DEX on the hepatic ER in this in vitro system are very similar to the effects of these hormones under in vivo conditions. The inducible expression of the ER has never before, to our knowledge, been demonstrated in any mammalian liver cell culture system. PMID- 8404594 TI - Human NCI-H295 adrenocortical carcinoma cells: a model for angiotensin-II responsive aldosterone secretion. AB - Excessive secretion of aldosterone from the adrenal results in the most common form of endocrine hypertension. An understanding of the regulatory processes involved in aldosterone synthesis and release is needed to define the biomolecular mechanisms controlling excessive production of aldosterone. However, in vitro studies regarding the regulatory mechanisms of human aldosterone production have been limited because of difficulties in obtaining tissue and the subsequent isolation of aldosterone-secreting glomerulosa cells. Herein we describe an adrenocortical carcinoma cell line, NCI-H295, which provides a suitable angiotensin-II (AII)-responsive model system to investigate the acute and chronic regulation of aldosterone synthesis. The cells were characterized with regard to the effects of AII on second messenger systems, aldosterone release, and levels of aldosterone synthase (P450c18) mRNA. In the presence of lithium, AII caused a rapid, but transient, increase in the production of inositol tris- and bisphosphates, whereas a prolonged gradual accumulation of inositol monophosphate occurred. Treatment with AII resulted in a 4.5-fold increase in total inositol phosphates in a concentration-dependent manner and an increase in intracellular cytoplasmic free Ca2+. Significant increases in aldosterone (3.5-fold) were detected within 1 h of AII addition. Aldosterone release occurred in a concentration-and time-dependent manner. The type 1 AII (AT1) receptor was shown to be responsible for activation of phosphoinositidase C, increased intracellular free Ca2+, and aldosterone production, as determined by use of the AT1 receptor antagonist DuP753. In addition, AII treatment resulted in a time-dependent increase in levels of P450c18 mRNA, as detected by RNAse protection assay. In summary, NCI-H295 cells provide a valuable model system to define mechanisms regulating human aldosterone production. PMID- 8404595 TI - Mechanism of pervanadate stimulation and potentiation of insulin-activated glucose transport in rat adipocytes: dissociation from vanadate effect. AB - Previous studies have shown that the combination of vanadate and H2O2 generates peroxide(s) of vanadate (pervanadate) that is able to mimic insulin in stimulating lipogenesis or protein synthesis and inhibiting lipolysis in rat adipocytes. Here we report that pervanadate is a potent trigger of 3-O methylglucose transport in rat adipocytes, with an effective concentration of 5 microM and a maximum at 20 microM. Moreover, pervanadate produced an additional activation of approximately 60% on glucose influx in cells treated with maximally activating concentrations of insulin. Vanadate was ineffective in potentiating insulin-stimulated glucose uptake. Quercetin, a bioflavonoid that inhibits insulin receptor tyrosine kinase, blunted this effect of pervanadate. Treatment of adipocytes with pervanadate inhibited protein phosphotyrosyl phosphatase activity of cell extracts in a dose-dependent manner, with an ID50 of 5 microM and complete inhibition at 80 microM. In contrast, vanadate (1-800 microM) did not appreciably inhibit cell phosphotyrosyl phosphatases. The inhibitory effect of pervanadate correlated with the increase in protein phosphotyrosine accumulation, as determined by Western blotting with antiphosphotyrosine antibodies. The most prominent phosphotyrosine-containing band detected in pervanadate-treated adipocytes was that of autophosphorylated insulin receptor, identified by immunoblotting or immunoprecipitation with antiinsulin receptor antibodies. The addition of insulin to pervanadate-treated adipocytes (20 microM) caused a further increase (approximately 70%) in receptor autophosphorylation. In a cell-free system using partially purified insulin receptor devoid of tyrosine phosphatase activity, pervanadate did not stimulate the receptor autophosphorylation or interfere with the stimulating effect of insulin. These results suggest that 1) pervanadate triggers glucose uptake by increasing autophosphorylation of insulin receptor, preventing its dephosphorylation; 2) under physiological conditions, cellular protein phosphotyrosyl phosphatase activity is high, thereby significantly opposing insulin-mediated hexose transport; and 3) pervanadate has the unique ability to markedly increase maximal cell responsiveness in stimulating glucose transport achieved at a saturating insulin concentration. These findings suggest a possible clinical application in the management of glucose uptake in pathological conditions of insulin resistance and hyperinsulinemia. PMID- 8404597 TI - Origin of neurohypophyseal neuropeptide-FF (FLFQPQRF-NH2). AB - Neuropeptide-FF (FLFQPQRF-NH2), originally isolated from bovine brain, is an FMRF NH2-like peptide with morphine-modulating activity. Neuropeptide-FF (NPFF) is unevenly distributed in the central nervous system, with the highest concentrations in posterior pituitary and spinal cord. In the rat pituitary, NPFF is found exclusively in the neural lobe, where it is localized in nerve terminals and fibers, indicating the hypothalamus as a possible source of the neural lobe NPFF. In this study the origin of neurohypophyseal NPFF was investigated using various hypothalamic lesions and an anterograde tracing experiment. The results suggest that at least part of the neurohypophyseal NPFF originates from the supraoptic nucleus and may be localized in some of the arginine vasopressin containing magnocellular neurons. PMID- 8404596 TI - Two gonadotropin-releasing hormones in the African catfish, Clarias gariepinus: localization, pituitary receptor binding, and gonadotropin release activity. AB - Two GnRH peptides have recently been identified in brain extracts of the African catfish, chicken-II GnRH ([His5,Trp7,Tyr8]GnRH, cGnRH-II) and catfish GnRH ([His5,Asn8]GnRH, cfGnRH). Using three experimental approaches, we investigated whether both peptides are involved in the regulation of pituitary gonadotropin secretion. First, the presence of cfGnRH and cGnRH-II in the pituitary was studied by biochemical and immunocytochemical techniques, as GnRH reaches the pituitary via axonal transport in teleost fish. Pituitary extracts contained cfGnRH- and cGnRH-II-immunoreactive material, showing the same HPLC retention times as the respective synthetic GnRH peptides; cfGnRH was present in 37-fold higher amounts than cGnRH-II. Using single and double labeling immunocytochemical techniques, both peptides were localized in the same peptidergic nerve fibers and often within the same secretory granules in the vicinity of the gonadotropes. Second, the two peptides were tested for their capacity to induce an increased secretion of the LH-like gonadotropin-II (GTH-II). In vivo studies showed that both GnRHs released GTH-II, but 100-fold higher cfGnRH than cGnRH-II doses were necessary to induce similar increases in circulating GTH-II levels. In vitro experiments using pituitary tissue fragments in a perifusion system also revealed a clearly higher GTH-II-releasing capacity of cGnRH-II compared to that of cfGnRH. Third, the peptides were tested for their ability to displace [125I]salmon GnRH analog ([D-Arg6,Trp7,Leu8,Pro9-NEt] GnRH, sGnRHa), a high affinity GnRH receptor ligand, from catfish pituitary membrane preparations. Chicken GnRH-II competed with [125I]sGnRHa for pituitary GnRH-binding sites, whereas cfGnRH did so only slightly. The present data show that cGnRH-II is the more potent GTH-II secretagogue, although a role for cfGnRH in the regulation of GTH-II secretion cannot be excluded. The high biological activity of cGnRH-II may be related to the regulation of GTH-II secretion surges, such as those associated with spawning, whereas cfGnRH may be involved in regulating moderate changes in GTH-II plasma levels. The peptides' potency differences appear to be related to their different binding affinities for the pituitary GnRH receptor. PMID- 8404598 TI - Insulin-like growth factor-I inhibits parathyroid hormone-stimulated and enhances prostaglandin E2-stimulated adenosine 3',5'-monophosphate production by human osteoblast-like SaOS-2 cells. AB - Insulin-like growth factor-1 (IGF-I), an endocrine and autocrine/paracrine factor that enhances collagen synthesis and bone matrix formation by osteoblasts, has been implicated in the coupling of bone formation with bone resorption. We have found, using SaOS-2 osteoblastic cells, that IGF-I inhibits PTH-stimulated cAMP production. Pretreatment of SaOS-2 cells with IGF-I for 24 h inhibited cAMP production stimulated by PTH with an IC50 of 1 nM and maximal inhibition to 10 20% of control values at an IGF-I concentration of 10 nM. Pretreatment with IGF-I had no effect on vasoactive intestinal peptide-stimulated cAMP production, but it enhanced cAMP production stimulated by prostaglandin E2 (PGE2) by as much as 5 fold at a concentration of 10 nM and with an EC50 of 1 nM. Pretreatment of SaOS-2 cells with IGF-I did not affect cholera toxin- or forskolin-stimulated cAMP accumulation. Taken together, these findings indicated that IGF-I did not affect Gs alpha, coupling of Gs alpha to adenylate cyclase, or adenylate cyclase itself. Binding experiments using [125I] chicken PTH-related peptide (PTHrP)-(1-36) [Tyr36]NH2 demonstrated that IGF-I reduced PTH/PTHrP receptor number to 25% of the control value without affecting receptor affinity. IGF-I and the related growth factors, insulin and IGF-II, inhibited PTH-stimulated cAMP production with a rank order of potency of IGF-I > or = IGF-II > insulin, indicating that the actions of IGF-I on SaOS-2 cells were probably mediated by the IGF-I receptor. We conclude that physiologically relevant concentrations of IGF-I specifically inhibited PTH-stimulated and enhanced PGE2-stimulated production of cAMP by an action at the level of PTH and PGE2 receptors and/or coupling of the receptors to Gs alpha. PMID- 8404599 TI - Functional properties of polyclonal antibodies raised against the N-terminus region (residues 9-30) of the follicle-stimulating hormone (FSH) receptor: significance of this receptor region in FSH recognition and signal transduction. AB - In this study, we raised polyclonal antibodies in rabbits against a synthetic peptide corresponding to a unique region of the FSH receptor, residues 9-30, with no sequence homology to receptors for LH and TSH, and examined their characteristics relevant to receptor function. Binding of [125I]human (h) FSH to membrane-bound receptors was inhibited in a concentration-dependent manner by the anti-FSH receptor-(9-30) peptide antibody. Preimmune serum had no effect. Lineweaver-Burke plot analysis of [125I]hFSH binding to membrane receptors in the presence or absence of antireceptor peptide antibody indicated that the antibody effectively competed with FSH at a hormone-binding site on the receptor. Also, antireceptor peptide antibody, but not preimmune serum, inhibited the ability of FSH to stimulate the conversion of androstenedione to estradiol in cultured immature rat Sertoli cells. Stimulation of estradiol synthesis by Sertoli cells caused by cholera toxin or forskolin (which are known to act through the Gs protein and catalytic unit of adenylate cyclase, respectively) was not inhibited by antireceptor peptide antibody. Indirect immunofluorescence staining of cultured rat Sertoli cells showed binding of antibody to plasma membrane receptor. No fluorescent staining of receptor was observed when cells were incubated with preimmune serum or antireceptor peptide antibody in the presence of excess receptor-(9-30) peptide or hFSH. These results were consistent with specific labeling of membrane-bound FSH receptors by anti-receptor-(9-30) peptide antibody. When detergent-solubilized membrane preparations from rat Sertoli cells were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions and then subjected to Western blot analysis, antireceptor peptide antibody, but not preimmune rabbit serum, specifically recognized intact FSH receptor. Although the antireceptor peptide antibody occupied the N-terminus 9-30 epitope region in the FSH receptor, it did not induce postbinding events, such as receptor patching (aggregation), as shown by indirect immunofluorescence staining of rat Sertoli cells and the estradiol response. In contrast, a polyclonal antibody against the FSH holoreceptor capable of interacting with multiple epitopes on the receptor could induce FSH-like effects, such as receptor patching and estradiol response in Sertoli cells. In conclusion, antibody raised against the N-terminus region (9-30) of the FSH receptor recognized intact FSH receptor, inhibited FSH binding, and behaved as an antagonist, suggesting that this N-terminus epitope region of the receptor is involved in hormone binding.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404600 TI - Evidence that the growth hormone receptor mediates differentiation and development of the mammary gland. AB - We have shown that nonlactogenic rat (r) GH is far more potent than rPRL in inducing rat mammary development. To determine the relative roles of GH and PRL in mammary development and their mechanisms of action, we have compared the abilities of a group of native and mutant GHs, PRLs, and placental lactogens (PLs) to induce mammary development, bind to GH receptors, and activate lactogenic receptors. Mammary development was assessed histologically by counting terminal end buds and alveolar structures in glands from sexually immature, hypophysectomized, castrated, estradiol-treated rats. Hormones were implanted, in Elvax pellets, into the lumbar mammary gland. Significant increases in terminal end buds (P < 0.03) over internal control values were obtained with rGH, recombinant human GH (rhGH), rbGH, and one of two mutant rhGHs. These four hormones were also found to bind to GH receptors with high affinity. In contrast, little development occurred with hPRL, rPRL, rhPL, ovine PRL, mutant forms of rhPRL and rhPL, and a mutant of rhGH altered to reduce binding to GH and PRL receptors. All of these substances are more than 50-fold reduced in binding to the GH receptor, yet can bind and activate lactogenic receptors. Thus, only those natural or mutant pituitary or placental hormones with high binding affinity to GH receptors induce mammary development, suggesting that GH receptors play a central role in this process. PMID- 8404601 TI - Locally produced angiotensin II induces ovulation by stimulating prostaglandin production in in vitro perfused rabbit ovaries. AB - The present study was undertaken to investigate the role of exogenous and endogenous angiotensin II (Ang II) in ovarian steroidogenesis and production of prostaglandin (PG) in in vitro perfused rabbit ovaries. The addition of 100 or 10 micrograms Ang II at 2-h intervals to the perfusate did not stimulate progesterone production, but significantly stimulated estradiol (E2) production by perfused rabbit ovaries. When the specific antagonist of Ang II, saralasin at 2 x 10(-6) M, was added to the perfusate 30 min before the onset of Ang II administration, Ang II-stimulated production of E2 was significantly blocked. Ang II also significantly stimulated both PGE2 and PGF2 alpha production, while the addition of saralasin to the perfusate significantly inhibited the Ang II stimulated production of PG. The levels of PGs in ovaries perfused with saralasin plus 100 micrograms Ang II did not differ significantly from those in control ovaries perfused with medium alone. Exposure to human CG (hCG) significantly stimulated production of progesterone and E2 by perfused rabbit ovaries, while the concomitant administration of 2 x 10(-6) M saralasin significantly reduced only E2 production. Addition of saralasin to the perfusate inhibited hCG stimulated PG production in a dose-dependent manner. The ovulatory efficiency in ovaries treated with hCG alone or hCG plus saralasin was significantly correlated with PG production by perfused rabbit ovaries at 12 h after exposure to hCG. The production of PG stimulated by Ang II was completely reduced by indomethacin treatment during the entire perfusion period. Indomethacin completely blocked Ang II-induced ovulation, but not Ang II-stimulated oocyte maturation. Concurrent administration of staurosporine, a protein kinase C inhibitor, at 10(-6) M significantly inhibited Ang II-stimulated meiotic maturation of ovulated ova and follicular oocytes. In conclusion, these results indicate that Ang II has a direct role in ovarian production of E2 and PG. An intrinsic renin-angiotensin system in the rabbit ovary may act as an intermediary of gonadotropin-stimulated PG production. Locally produced Ang II may induce ovulation in the rabbit ovary, at least in part, by stimulating PG production. PMID- 8404602 TI - Identification by analytical flow cytometry of prolactin receptors on immunocompetent cell populations in the mouse. AB - Five monoclonal antibodies (T1, T6, U5, U6, and E21) made to the external portion of the rat PRL receptor (PRL-R) were conjugated to fluorescein isothyrocynate and used to examine the presence of PRL-R on mouse lymphocytes and macrophages using analytical flow cytometry. The monoclonals were initially evaluated using Nb2 cells, a cloned line from a rat lymphoma, and NOG-8 cells, a cloned line from normal mouse mammary gland tissue, which are known to have PRL-R. All monoclonal antibodies bound to these cells, but the U5 monoclonal gave the best separation from unstained cells. CTLL-2 cells, a mouse lymphoma cell line containing interleukin-2 receptors, did not bind to any of the monoclonals. Isolated thioglycolate-induced peritoneal macrophages contained PRL-R, and the PRL-R monoclonal U5 gave the best separation from unstained cells. Eighty-five percent of macrophages constitutively had PRL-R using this monoclonal. In vivo stimulation of the popliteal lymph node by the injection of Concanavalin-A (Con A) into the right foot pad of intact and ovariectomized (OVX) BALB/c mice induced, at the end of 10-12 h, a marked increase in interleukin-2 receptor (IL 2R) expression on CD4, CD8, and B-cells compared to the unstimulated left popliteal lymph node. The number of CD4 and CD8 cells from OVX animals with IL-2R was twice that from intact animals, whereas no difference in the percentage of B cells with IL-2R was evident. PRL-R were constitutively expressed on 5% of the CD4 cells and 20% of the CD8 cells and were increased in the Con-A-stimulated lymph node when examined with the U5 PRL-R monoclonal. A higher percentage of CD4 and CD8 cells from OVX animals constitutively expressed PRL-R, and when stimulated with Con-A, a further increase was observed compared to the level in intact animals. Using the U5 monoclonal, over 80% of the B220 cells constitutively expressed PRL-R; however, when T1, T6, and U6 monoclonals were examined, the percentage was considerably below (20%) than that observed for U5. Con-A stimulation did not alter the percentage of B220 cells expressing PRL-R. These results show the importance of identifying lymphocyte subsets and examining a number of PRL-R monoclonals in determining lymphocyte PRL-R expression on the surface of the cell. PMID- 8404603 TI - Variable patterns of gonadotropin-releasing hormone secretion during the estrogen induced luteinizing hormone surge in ovariectomized ewes. AB - There have been a number of studies of GnRH secretion into the hypophysial portal blood at the time of the estrogen-induced LH surge, but the exact pattern of secretion at the onset of the positive feedback event remains a point of some dispute. In the present study, GnRH concentrations in portal plasma and jugular venous LH concentrations were measured in samples taken at 2.5-min intervals from five ovariectomized control ewes and seven ovariectomized ewes that were treated with 50 micrograms estradiol benzoate (im) to cause a LH surge. The frequency of GnRH pulses was calculated using a modification of previous criteria because of the frequent sampling. It was found that GnRH pulse frequency increased during the LH surge, being greatest during the "leading edge" of the surge. Perhaps more importantly, the data show that variable patterns of GnRH secretion occur at the start of the LH surge and during the surge, suggesting that no single pattern can be identified as an initiation signal of the positive feedback event. These data are consistent with other results that have been obtained in this laboratory in cyclic animals. PMID- 8404604 TI - In vitro characterization of gonadotropin-releasing hormone antagonists in goldfish, Carassius auratus. AB - The two native forms of GnRH, salmon GnRH and chicken GnRH-II, in the brain and pituitary of goldfish are both active in stimulating gonadotropin-II (GTH-II) and GH release. The objective of the present study was to characterize GnRH antagonists for their ability to inhibit sGnRH- and cGnRH-II-induced GTH-II and GH release in goldfish using a pituitary fragments perifusion system. Contrary to expectations, putative GnRH antagonists with D-Arg6 stimulated GTH-II and GH release in nearly all cases. [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]mammalian (m) GnRH inhibited sGnRH- and cGnRH-II-stimulated GTH-II release in a dose-dependent manner, with ED50 values of 242 +/- 48 and 169 +/- 17 nM, respectively. [Ac-delta 3-Pro1,4FD-Phe2,D-Trp3,6]mGnRH also inhibited GH release stimulated by sGnRH (ED50, 128 +/- 74 nM) and cGnRH-II (ED50, 157 +/- 67 nM). The degree of inhibition was higher in sexually regressed fish compared to postspawning fish. [D-p-Glu1,D-Phe2,D-Trp3,6]mGnRH suppressed both sGnRH- and cGnRH-II-induced GTH II release with ED50 values of 326 +/- 96 and 249 +/- 74 nM, respectively. [Ac delta 3-Pro1,4FD-Phe2,D-Trp3,6]sGnRH inhibited sGnRH and cGnRH-II stimulated GTH II release, but stimulated GH release. On the other hand, [Ac-D(2)-Nal1,4Cl-D Phe2,D-(3)Pal3,6,Arg5,D-A la10]mGnRH weakly stimulated GTH-II release, but strongly inhibited basal GH release. These results indicate that [Ac-delta 3 Pro1,4FD-Phe2,D-Trp3,6]mGnRH has clear antagonistic activity on sGnRH and cGnRH II stimulation of GTH-II and GH release in vitro. The differential actions of a few GnRH analogs on GTH-II and GH release indicate that the properties of the GnRH receptors on GTH and GH cells may be different. The amino acid in position 6 plays an important role in determining the nature of intrinsic activity of GnRH peptides, and substitution of D-Arg6 normally produces agonistic analogs. PMID- 8404605 TI - Extrahepatic expression of fibrinogen by granulosa cells: potential role in ovulation. AB - Granulosa cells from ovarian follicles were shown to express and secrete fibrinogen under the control of FSH. Conditioned medium was collected from granulosa cell cultures and found to contain FSH-dependent 50-kilodalton (kDa) and 93- to 95-kDa proteins. N-Terminal microsequence analysis identified these proteins as fibrinogen beta- and gamma-chains, respectively. Proteins migrating at 93 and 95 kDa contain identical gamma-chain sequences at the N-terminal, suggesting differential processing of fibrinogen. These fibrinogen chains were specifically detected with antifibrinogen antibodies in immunoblot and immuno precipitation analysis. Fibrinogen gamma-chain mRNA was detected in granulosa cells by polymerase chain reaction analysis, confirming fibrinogen gene expression by these cells. Fibrinogen secretion by granulosa cells was measured by a competitive enzyme-linked immunosorbent assay. Granulosa cells treated with FSH (100 ng/ml) secreted 2-3 times more fibrinogen than untreated cells. These data show that fibrinogen, a major product of the liver, is also a secretory product of granulosa cells. This provides a novel extrahepatic site of fibrinogen expression. As hepatic parenchymal cells normally maintain high circulating levels of fibrinogen, the local production of fibrinogen in the ovary is anticipated to have specialized functions. Locally produced fibrinogen may be important in the clotting process following tissue rupture at ovulation. In addition, fibrinogen fragments may be involved in the mechanism of ovulation by increasing the activity of tissue-type plasminogen activator to control the proteolytic activity required for ovulation. PMID- 8404606 TI - Preovulatory gonadotropin-releasing hormone surge in ovarian-intact rhesus macaques. AB - The occurrence and profile of the preovulatory hypothalamic GnRH surge in relation to plasma profiles of LH and ovarian steroids, i.e. 17 beta-estradiol (E2) and progesterone (P4), were examined in ovarian intact, freely moving rhesus macaques. Nine monkeys with active ovarian cycles were each fitted with a jugular venous catheter and two push-pull cannulae directed to separate sites within the median eminence (ME). Each female was connected continuously to a tether/swivel device through which daily blood samples or frequent blood samples and ME perfusates (simultaneously at 10- to 20-min intervals for 18-24 h) were obtained without disturbing the animals. An increment in the plasma E2 level (> 150 pg/ml) during the follicular phase (FP) was selected as the preovulatory ovarian signal and served as the index for initiating the ME push-pull perfusion (PPP). Daily increased P4 concentrations of more than 1 ng P4/ml plasma for several consecutive days were consistent with the assumption of ovulation and subsequent formation of a corpus luteum after PPP. A total of 18 PPP trials were completed; each in a fresh ME site. Five of these PPPs were performed during the mid- and late FP (3 were between 6-8 days before and 2 were 4 days before the E2 peak). The remaining 13 PPPs, each of 18- to 24-h duration, were performed between 24 h before and 48 h after the highest daily plasma E2 level, i.e. time zero. Of these 13 PPPs, 2 started within 12 h before (-12 to 0 h), 8 began within 12 h after (0 12 h), and 3 started between 12-24 h after this peak E2 value. During the FP, mean levels of GnRH and LH were less than 2 pg/ml and 20 ng/ml, respectively. During the periovulatory interval (-24 to 48 h around time zero), the release of hypothalamic GnRH (expressed in picograms per ml) increased to 6.63 +/- 2.35 between -12 to 0 h (n = 2), peaked at 20.70 +/- 6.09 between 0-12 h (n = 10), declined to 3.25 +/- 1.39 between 12-24 h (n = 11), and further declined to 0.89 +/- 0.18 between 24-36 h (n = 3). The mean GnRH value from 0-12 h was higher (P < 0.05) than other means (including those during the FP), except for the value between -12 to 0 h. Changes in mean plasma LH values during the same periods paralleled those in GnRH.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404607 TI - Effects of activin-A, inhibin-A, and transforming growth factor-beta 1 on stage specific deoxyribonucleic acid synthesis during rat seminiferous epithelial cycle. AB - Activin and inhibin are members of the transforming growth factor-beta (TGF beta) gene family. They are expressed in various organ systems, where they possess regulatory functions. Inhibin, activin, and TGF beta have been reported to also be expressed in the adult rat testis. We studied in vitro the action of these growth factors on premitotic and premeiotic DNA synthesis during the rat seminiferous epithelial cycle. Two-millimeter rat seminiferous tubule segments were isolated by transillumination-assisted microdissection from stages V, VIIa, VIII-IX, and I of the cycle and incubated in vitro in the presence of activin-A, inhibin-A, or TGF beta 1. During 24-, 48-, and 72-h incubation spontaneous progression of spermatogenesis was noted. The staged samples allowed us to selectively quantitate DNA synthetic activity of specific germ cell types. At the end of the culture, the tubules were pulse labeled with [3H]thymidine, and DNA synthesis was quantified by liquid scintillation counting, and the activated cells were detected by autoradiography. Activin-A stimulated preleptotene spermatocyte DNA synthesis in a dose-dependent manner. DNA synthesis of intermediate spermatogonia was also stimulated by activin-A, whereas inhibin-A inhibited DNA synthesis of these cells. TGF beta 1 had a small, but significant, stimulatory effect on DNA synthetic activity at stage VII. These results support the view that activin-A, inhibin-A, and TGF beta 1 take part in the regulation of DNA synthesis during rat spermatogenesis. PMID- 8404608 TI - Bovine adrenal glomerulosa and fasciculata cells exhibit 28.5-kilodalton proteins sensitive to angiotensin, other agonists, and atrial natriuretic peptide. AB - Protein synthesis by bovine adrenal glomerulosa and fasciculata cells in response to various modulators of steroid synthesis was examined using [35S]methionine labeling and two-dimensional gel electrophoresis. Both cell types responded to steroidogenic stimuli with rapid changes in a family of 28- to 30-kilodalton (kDa) proteins similar to those described in rat fasciculata by Epstein and Orme Johnson. In glomerulosa, angiotensin-II (AII), potassium, and (Bu)2cAMP stimulated the appearance of two 28.5-kDa proteins (no. 3 and 4) with pI values of 6.44 and 6.33 and decreased labeling of two other 28.5-kDa proteins (no. 1 and 2) with pI values of 6.9 and 6.59. The rank order of potency on aldosterone synthesis and that on proteins 1-4 were the same: (Bu)2cAMP > AII > potassium. Atrial natriuretic peptide blocked the effects of AII on all four proteins and on aldosterone synthesis. Adrenal secretagogues also affected labeling of four slightly larger (30 kDa) proteins (no. 5-8). Corresponding proteins in each quartet are separated by the same difference in isoelectric points. These eight proteins may represent a core protein systematically modified in a number of ways. Aldosterone synthesis in glomerulosa, like glucocorticoid synthesis in fasciculata, requires ongoing protein synthesis. The 28- to 30-kDa proteins increased by steroidogenic stimuli in both cells and decreased by atrial natriuretic peptide in glomerulosa may be the proteins whose synthesis is crucial to acute control of steroidogenesis. Our results indicate that these proteins are made in response to calcium- or calcium/phosphoinositide-dependent mechanisms as well as by cAMP. PMID- 8404609 TI - The type 1 angiotensin-II receptor mediates intracellular calcium mobilization in rat luteal cells. AB - The intracellular cytosolic calcium concentration ([Ca]i) was determined in cultured rat luteal cells using the calcium-chelating dye fura-2 and microspectrofluorimetry. Angiotensin-II (Ang-II) induced a dose-dependent transient increase in [Ca]i (ED50, 9.0 +/- 6.5 nM). After the initial peak in [Ca]i, cytosolic calcium returned to a secondary elevated basal level that was dependent upon the presence of extracellular calcium. Pretreatment of rat luteal cells with Ang-II (100 nM) desensitized a subsequent response to a higher concentration (1 microM), but did not desensitize a prostaglandin F2 alpha (PGF2 alpha)-induced calcium flux. Although the peak increases in [Ca]i induced by Ang II (1 microM) and PGF2 alpha (10 microM) were not significantly different, the plateau phase stimulated by PGF2 alpha was significantly higher (P < 0.05) than that stimulated by Ang-II (1 microM). Pretreatment of luteal cells with the type 2 Ang-II receptor antagonist PD 123319 (10 microM) did not inhibit calcium mobilization; however, Ang-II (1 microM)-induced calcium mobilization was dose dependently blocked by the type 1 Ang-II receptor antagonist Losartan (DuP 753). The ID50 for Losartan was 5.2 +/- 1.8 nM. Pretreatment of the luteal cells with the endoplasmic reticulum calcium ATPase inhibitor thapsigargin (1 microM) also blocked Ang-II-induced calcium mobilization. These data demonstrate the presence of the type 1 Ang-II receptor in rat luteal cells, through which Ang-II dose dependently mobilizes calcium from an intracellular source, probably the endoplasmic reticulum. PMID- 8404610 TI - Glucagon-like peptide-1-(7-36)amide and a rise in cyclic adenosine 3',5' monophosphate increase cytosolic free Ca2+ in rat pancreatic beta-cells by enhancing Ca2+ channel activity. AB - Glucagon-like peptide-1 (GLP-1), in the form of either GLP-1-(7-36)amide or GLP-1 (7-37), has been shown to potently stimulate insulin release in a glucose dependent manner and is suggested to be a physiological incretin. To explore the mechanisms by which GLP-1-(7-36)amide stimulates insulin release, we investigated its action on the cytosolic free Ca2+ concentration ([Ca2+]i) in single rat pancreatic beta-cells by the dual wavelength microfluorometry with fura-2. In the presence of 8.3 mM glucose, GLP-1-(7-36)amide at a concentration as low as 3 x 10(-12) M produced a rapid transient increase in [Ca2+]i in some of the single beta-cells. GLP-1-(7-36)amide at 10(-11) M or more evoked the [Ca2+]i response in the majority of beta-cells. In the presence of 2.8 mM glucose, GLP-1-(7-36)amide was without effect. The [Ca2+]i response to GLP-1-(7-36)amide was completely and reversibly inhibited under Ca(2+)-free conditions and by 1 microM nitrendipine, a blocker of L-type Ca2+ channels. Elevation of cAMP in beta-cells by either 10 microM forskolin, an activator of adenylyl cyclase, or 5 mM (bu)2cAMP (db-cAMP) produced an increase in [Ca2+]i similar to that caused by GLP-1-(7-36)amide. The db-cAMP-induced increase in [Ca2+]i was also completely blocked by nitrendipine. In the continuous presence of GLP-1-(7-36)amide and after the transient [Ca2+]i increase it elicited, db-cAMP failed to evoke the [Ca2+]i response. It is concluded that GLP-1-(7-36)amide at physiological concentrations and a rise in cAMP increase [Ca2+]i in pancreatic beta-cells by enhancing the activity of L type Ca2+ channels in the beta-cell plasma membrane. It is suggested that the cAMP-operative mechanism is involved in the GLP-1-(7-36)amide action to increase [Ca2+]i in beta-cells. PMID- 8404611 TI - Insulin attenuation of vasopressin-induced calcium responses in arterial smooth muscle from Zucker rats. AB - Insulin attenuates agonist-induced vascular contractility of aortic rings and decreases vasopressin (AVP)-elicited increases in vascular smooth muscle cell (VSMC) intracellular calcium ([Ca2+]i). To determine if insulin's effects on AVP induced [Ca2+]i responses are altered in an insulin-resistant and hypertensive state, we studied vascular smooth muscle calcium responses in VSMC derived from Zucker lean and obese rats. AVP concentration-response experiments revealed that VSMC derived from obese animals exhibited exaggerated [Ca2+]i responses over the range of 1 x 10(-10) to 1 x 10(-7) M AVP compared to lean controls (P < 0.05, by multiple analysis of variance). Insulin treatment (7 x 10(-7) M) decreased the [Ca2+]i response to 1 nM AVP by 66 +/- 8% and 71 +/- 9% in lean and obese VSMC, respectively. Similar decreases were observed with the 10 nM AVP stimulus (41 +/- 9% and 61 +/- 7%, for lean and obese, respectively). AVP receptor binding studies revealed that exaggerated [Ca2+]i responses in obese VSMC were not due to alterations in AVP-binding properties (no significant differences in ID50, Kd, or binding capacity in lean and obese VSMC preparations). In addition, insulin treatment (1 x 10(-7) M) resulted in no differences in AVP receptor-binding properties in either cell line. Therefore, exaggerated [Ca2+]i responses in obese VSMC are most likely due to a postreceptor abnormality. These abnormalities in VSMC [Ca2+]i metabolism preceed and may play a role in the development of hypertension in the Zucker obese rat. Although insulin resistance in Zucker obese rats has been demonstrated in several tissues, VSMC [Ca2+]i responses to AVP are, nonetheless, similarly attenuated by insulin in obese and lean VSMC preparations. PMID- 8404612 TI - Thyrotropin-releasing hormone stimulates c-jun and c-fos messenger ribonucleic acid levels: implications for calcium mobilization and protein kinase-C activation. AB - The hypothalamic neuropeptide TRH, through G-protein-coupled transmembrane pituitary receptors, rapidly stimulates intracellular signaling events that, in turn, stimulate gene transcription. Our previous studies in transfected pituitary tumor cells indicated that TRH stimulation of thyrotropin beta-subunit (TSH beta) gene expression involves both calcium mobilization and protein kinase-C activation. To characterize the gene-proximal elements of the intracellular signaling pathways involved, we examined the effects of TRH, ionomycin, and phorbol ester (TPA) on cellular protooncogenes (c-jun and c-fos) known to be responsive to calcium mobilization and protein kinase-C activation. TRH stimulated a 3-fold increase in both c-jun and c-fos mRNA levels within 1 h, followed by a rapid decline in steady state mRNA levels. A secondary response to the single administration was noted, culminating in a 5-fold stimulation at 20 h. The increase in c-jun and c-fos mRNA levels occurred before the increased steady state mRNA levels of both PRL and TSH beta chimera in transfected pituitary GH3 cells. Furthermore, we examined the role of calcium in these effects using the ionophore ionomycin to elevate and TMB-8 to decrease intracellular calcium. We used the phorbol ester TPA to investigate the effects of increased protein kinase C activity and H7 or pretreatment with TPA to monitor the decreased kinase activity. Our data indicate that calcium mobilization and protein kinase activation represent distinct components of the signaling events initiated by TRH resulting in increased c-jun and c-fos mRNA levels. Only c-fos mRNA is increased by all three factors, suggesting that c-fos may be a key element in mediating the intracellular processes reflecting TRH action. PMID- 8404613 TI - Regulation of pituitary corticotropin releasing hormone (CRH) receptors by CRH: interaction with vasopressin. AB - The physiological regulation of ACTH secretion is largely dependent on the interactive effects of CRH and vasopressin (VP) in the pituitary corticotroph. The importance of the magnocellular and parvicellular hypothalamic systems as a source of VP for pituitary regulation was studied by analyzing the effects of endogenous activation of these systems or VP infusion on the ability of CRH to regulate its receptor sites in the anterior pituitary. CRH receptors were measured by binding of [125I]Tyr-ovine CRH to membrane-rich fractions of anterior and neurointermediate pituitary lobes. Minipump infusion of 100 ng/min CRH for 48 h caused a 40% decrease in the anterior pituitary CRH receptor concentration. Simultaneous infusion of VP markedly potentiated the effect of CRH, but only at doses that elevated plasma VP to levels in the range of those in the pituitary portal circulation. Activation of the magnocellular vasopressinergic system by 60 h of water deprivation increased plasma VP levels from 0.5 +/- 0.1 to 11.8 +/- 0.6 pg/ml, but had no effect on the anterior pituitary CRH receptor concentration in control rats or animals receiving CRH infusion (100 ng/min for 48 h). The CRH receptor concentration was significantly increased in neurointermediate pituitary membranes from water-deprived rats. When the parvicellular vasopressinergic system was activated by 14 days of repeated restraint stress, there was a significant enhancement in the ability of CRH to decrease anterior pituitary CRH receptors (33% and 62% in control and stressed rats, respectively). The concomitant infusion of 200 ng/min of the VP antagonist [(mercapto cyclopentamethylene propionic acid)-[methyl-tyrosine]arginine VP] during the CRH infusion in chronically stressed rats significantly reduced the magnitude of the pituitary CRH receptor loss from a 62% to a 43% decrease (P < 0.01). In conclusion, exogenous VP modulates pituitary CRH receptor regulation by CRH only at doses sufficiently high to provide peripheral VP concentrations in the range of the circulating levels in hypophysial-portal blood. Furthermore, the demonstration that chronic endogenous activation of the parvicellular, but not the magnocellular, vasopressinergic system enhances the down-regulatory effect of CRH on anterior pituitary CRH receptors is in support of a critical role of parvicellular VP in the control of the corticotroph function. PMID- 8404614 TI - Pretranslational regulation of type I 5'-deiodinase by thyroid hormones and in fasted and diabetic rats. AB - In both humans and experimental animals, nutritional deprivation and systemic illness are associated with decreases in circulating T3 levels, T3 production, and type I 5'-deiodinase (5'DI) activity. In order to assess the regulation of 5'DI messenger RNA (mRNA) in these conditions, a solution hybridization assay was developed which utilized a complementary (cDNA) encompassing 92% of the 5'DI coding region. The administration of T3 to hypothyroid rats induced a 50-fold increase in 5'DI mRNA levels that preceded by 12 h a similar increase in 5'DI activity. Fasting for 48-72 h, but not 24 h, was associated with an approximate 50% decrease in 5'DI mRNA levels in liver, which preceded a decrease of similar magnitude in 5'DI activity. Fasting in hyperthyroid rats did not lower hepatic 5'DI mRNA levels or activity, nor did fasting attenuate the stimulatory effects of T3 administration on these parameters. In diabetic animals 5'DI mRNA levels and activity were markedly reduced. These results demonstrate that regulation of 5'DI by thyroid hormones and in fasting and diabetes mellitus is exerted principally at the pretranslational level. The relatively late occurrence of decreases in 5'DI mRNA levels and activity observed in nutritional deprivation suggests that these changes are secondary to, rather than causative of, the decreased serum and tissue T3 levels noted in these conditions. PMID- 8404615 TI - Retinoic acid potentiates phorbol ester-mediated induction of urokinase and plasminogen activator inhibitor type 2 in human myeloid leukemic cell lines. AB - We investigated the interactive regulation of the plasminogen activators (PAs) and their inhibitors (PAIs) by all-trans-retinoic acid (RA) in the presence and absence of the phorbol ester, phorbol myristate acetate (PMA), in four developmentally distinct human myeloid leukemic cell lines. Treatment of HL-60, K562, THP-1, and U937 cells with PMA resulted in an induction of urokinase-type PA (u-PA), the u-PA receptor (u-PAR), and PAI types 1 and 2 (PAI-1 and PAI-2). The addition of RA alone failed to alter gene expression or antigen production of PAI-1, PAI-2, or u-PAR. However, RA potentiated PMA-mediated induction of PAI-2 mRNA in HL-60 and U937 cells and PAI-2 antigen in all four cell lines. The effect of PMA on u-PA mRNA was also potentiated by RA in HL-60 and U937 cells. A similar, but transient, effect was seen on u-PA antigen levels. Run-on transcription analysis confirmed that these effects were due at least in part to changes in gene template activity. Furthermore, RA did not potentiate the effects of PMA on either u-PAR or PAI-1. In fact, in U937 cells, RA inhibited PMA-induced PAI-1 antigen secretion by approximately 60%. It would seem that interactive regulation of these genes allows for greater diversity of control, which may, in turn, be required for localized control of plasminogen-dependent extracellular proteolysis generated by monocytes/macrophage during cell migration and tissue remodeling. PMID- 8404616 TI - Transforming growth factor-alpha (TGF alpha) concentrations increase in regenerating rat liver: evidence for a delayed accumulation of mature TGF alpha. AB - Changes in the concentration of transforming growth factor-alpha (TGF alpha) protein were measured in regenerating liver. TGF alpha stimulates both DNA and protein synthesis in various liver-derived cells, and its mRNA levels increase in liver after partial hepatectomy (PH), suggesting that it may be an important autocrine regulator of liver regeneration. Using a sheep antiserum raised against mature rat TGF alpha, we developed a sensitive TGF alpha RIA. TGF alpha was extracted from livers in a detergent-containing buffer with protease inhibitors. Liver extracts, to a volume of 10 microliters/tube, produced a displacement curve of [125I]TGF alpha that was parallel to the pure standard. The TGF alpha content of normal liver was 57.04 +/- 26.25 pg/mg protein, 5.24 +/- 2.61 ng/mg DNA, and 10.33 +/- 4.47 ng/g liver (n = 5; mean +/- SD). Between 13-17 h after operation, TGF alpha concentrations in the livers of PH animals increased over those in sham operated (SH) controls (P < 0.05) and remained twice those in SH controls for more than 96 h, returning to control values by 8 days. In unoperated liver, gel chromatography showed all TGF alpha immunoactivity to be in fractions corresponding to known TGF alpha precursors (15-30 kilodaltons). Mature 5.6 kilodalton TGF alpha was not detected until 48 h after PH and was still present at 96 h. These data support a role for TGF alpha in the response to PH in the rat. However, the presence of TGF alpha precursors in normal liver, the short (< 4-h) interval between the increase in TGF alpha concentrations and the onset of hepatocyte DNA synthesis, the sustained elevation of TGF alpha levels after DNA synthesis has ceased, and the lack of detectable processing to the mature form until DNA synthesis has subsided all suggest that the membrane-anchored precursor and the mature forms of TGF alpha may have different functions, cellular sources, or target cells in regenerating liver. PMID- 8404617 TI - Cloning of four growth hormone/chorionic somatomammotropin-related complementary deoxyribonucleic acids differentially expressed during pregnancy in the rhesus monkey placenta. AB - The close homology of the chorionic somatomammotropin (CS) genes with pituitary GH is unique to primates. Southern blots of rhesus genomic DNA probed with a human CS cDNA demonstrated that the rhesus gene family consists of at least five EcoRI fragments between 2.3-9.5 kilobases. Rhesus pituitary and placental cDNA libraries were screened for hCS-hybridizing clones. A pituitary GH cDNA was isolated that was 95% and 96% identical to human GH at the mRNA and protein levels, respectively. Thirteen placental clones representing four different cDNAs were identified and sequenced. The relationships of the rhesus placental clones among themselves were similar to those among human placental mRNAs. Two cDNAs (mCS1 and mCS2) differed by two bases and coded for identical proteins. A third cDNA (mCS3) was 94% homologous to mCS1/2 in mRNA sequence as well as in deduced amino acid sequence. A fourth cDNA (mGH-V) was 84-86% homologous to the other placental cDNAs at the mRNA and protein levels, similar to the homology between human placental GH-V and CS mRNAs. To determine the relative expression of placental mRNAs, we developed a quantitative reverse transcription-polymerase chain reaction assay using diagnostic NlaIV restriction enzyme sites in cDNAs amplified from the placental mRNAs. All four mRNAs were expressed in the placenta throughout most of pregnancy, with the mCS1 and mGH-V mRNAs generally expressed at 2- to 10-fold higher levels than mCS2 and mCS3. Additionally, mCS3 and mGHV were expressed at relatively higher levels during the second and third trimesters than during the first trimester. PMID- 8404618 TI - Chronic intracerebroventricular neuropeptide-Y administration to normal rats mimics hormonal and metabolic changes of obesity. AB - Chronic intracerebroventricular (icv) administration of neuropeptide-Y (NPY; 10 micrograms/day) was performed in normal female rats to investigate its hormonal and metabolic consequences. Intracerebroventricular NPY produced hyperphagia, increased basal insulinemia, as well as liver and adipose tissue lipogenic activity. It also increased basal morning corticosteronemia. When NPY-induced hyperphagia was prevented by pair-feeding, the icv NPY treatment resulted in the same increases in basal insulinemia and corticosteronemia, and liver and white adipose tissue lipogenesis was still higher than that in respective controls. Under the ad libitum and pair-feeding conditions, icv NPY stimulated glucose uptake as well as total lipoprotein lipase activity in white adipose tissue; it resulted in an increase total activity of hepatic and white adipose tissue acetyl coenzyme-A-carboxylase. As all hormonal and metabolic changes elicited by icv NPY remained present (at the same or to a lesser extent depending upon the parameter considered) when hyperphagia was prevented by pair-feeding, it was, thus, shown that icv NPY per se induces peripheral hormonal and metabolic alterations via efferent routes, which remain to be determined. The effects of icv NPY reported in this study are similar to the defects observed in the early phase of genetic obesity in rodents, the hypothalamus of which has increased NPY levels. NPY could, thus, be of relevance in the occurrence of genetically induced obesity. PMID- 8404619 TI - Natriuretic hormone receptors and actions on bone endothelial cells. AB - [125I]Atrial natriuretic peptide (ANP) was used to identify ANP receptors on a clonal line of bovine bone endothelial (BBE) cells. Specific binding of [125I]ANP was saturable and of high affinity. Computer analysis of the equilibrium binding data indicated that the Scatchard plots are best fit by a straight line (Kd = 69.3 +/- 20.9 pM; binding capacity = 37.9 fmol/10(6) cells). The order of potency for competing with [125I]ANP binding was human ANP (hANP) > rat atriopeptin-1 (rAP-1) > porcine brain natriuretic peptide (pBNP) > porcine C-type natriuretic peptide. Affinity cross-linking studies indicated the presence of two major 130- and 70-kilodalton bands that specifically bound to hANP, rAP-1, pBNP, and porcine C-type natriuretic peptide. The binding of natriuretic peptides to BBE cells resulted in an increase in cGMP production and a significant decrease in Na+/K+/Cl- cotransport, without effects on cAMP intracellular accumulation. hANP, rAP-1, and pBNP at 100-nM concentrations, significantly inhibited PTH-induced cAMP production. Treatment with natriuretic hormones was also associated with an increase in 6-keto-prostaglandin F1 alpha levels in the culture medium of BBE cells and a higher cell growth rate. These studies demonstrate that bone endothelial cells bear receptors for natriuretic hormones associated with changes in PTH-induced cAMP production, prostaglandin production, and cell proliferation. PMID- 8404620 TI - Epidermal growth factor binding in the developing male reproductive duct and its regulation by testosterone. AB - It was shown previously that epidermal growth factor (EGF) plays a role in the testosterone-dependent fetal Wolffian duct differentiation. In this communication, the role of EGF was further investigated by determining whether EGF receptor plays a role in mediating the effect of EGF during Wolffian duct (male duct) differentiation, and experiments were designed to test this hypothesis. EGF-binding activity in cultured cells isolated from the reproductive ducts of 14- to 18-day-old fetal mouse was determined, and the results were evaluated by saturation/Scatchard analysis. Using this analysis, we demonstrate that the fetal mouse Wolffian duct contains EGF-saturable EGF-binding proteins with a binding affinity of 10(-10) M. The binding was temperature, time, and cell concentration dependent. Affinity cross-linking analysis of EGF binding of this tissue indicated the presence of an EGF receptor protein of 150 kilodaltons mol wt. The binding activity increased in parallel with the progression of Wolffian duct differentiation during the 14th to 18th day of gestation, which is the critical period of Wolffian duct differentiation. The binding activity was barely detectable at the onset of differentiation, i.e. on the 14th day of gestation. Binding activity in the male duct was higher than that in the female duct. Prenatal administration of testosterone during days 13-17 of gestation increased the EGF receptor concentration in the masculinized female fetuses, suggesting a role of fetal testicular testosterone in determining EGF-binding activity. Prenatal treatment of cyproterone acetate (an antiandrogen at the level of androgen receptor binding to androgen), however, produced no effect on EGF binding activity in male fetuses. Thus, a role for the EGF receptor is indicated during Wolffian duct differentiation. Testosterone appears to play a role in modulating EGF-binding activity. Further work is necessary to determine the mechanism by which testosterone modulates EGF-binding activity in this tissue. PMID- 8404621 TI - Rat testicular myoid cells respond to endothelin: characterization of binding and signal transduction pathway. AB - The presence of endothelin (ET), a vasoconstrictor peptide, in the testis suggests that it may regulate nonvascular target cells. We investigated binding ability, regulation of inositol phosphate metabolism, changes in cytosolic free Ca2+ concentrations ([Ca2+]i), and induction of morphological changes by ET-1 in rat primary testicular myoid cell cultures. ET-1 specifically bound to highly purified rat testicular myoid cells in a time- and temperature-dependent manner. Scatchard analysis of the binding studies indicated the presence of a single class of high affinity binding sites, with an apparent Kd of 3 x 10(-10) M and a maximal binding capacity of 10(5) sites/cell. ET-1 induced both rapid production of inositol triphosphate and mobilization of [Ca2+]i in a concentration-dependent fashion. By contrast, inositol lipid metabolism was slightly affected by ET-1 in the total peritubular cell population. Purified Sertoli cells failed to show either ET-1 binding or ET-1-induced phosphatidylinositol hydrolysis. Mobilization of [Ca2+]i mostly depended upon the release of Ca2+ from thapsigargin-sensitive intracellular Ca2+, whereas it was not affected by abolishment of the Ca2+ gradient through the plasma membrane or by inhibition of L-type voltage-sensitive Ca(2+)-channels by nifedipine. These findings together with the fact that Sertoli cells are unable to respond to and bind ET-1 indicate that ET is a specific agonist of myoid cells in the seminiferous tubule and suggest a role for ET-1 in the autocrine/paracrine regulation of testicular function. PMID- 8404622 TI - Streptavidin blotting: a sensitive technique to study cell surface proteins; application to investigate autophosphorylation and endocytosis of biotin-labeled insulin receptors. AB - Covalent attachment of biotin provides a useful method to label cell surface proteins. Subsequent to biotinylation, the protein can be purified by immunoprecipitation with a specific antibody, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. After transfer to a membrane by electroblotting, the biotinylated protein can be detected by probing with labeled streptavidin. This technique has been used to investigate recombinant human insulin receptors expressed on the surface of murine NIH-3T3 cells. Biotinylation of the extracellular domain with an impermeant reagent did not impair the ability of an antibody directed against an epitope in the intracellular domain to immunoprecipitate insulin receptors. In contrast, biotinylation reduced the avidity of a polyclonal antibody directed against the extracellular domain of the receptor. Nevertheless, by increasing the concentration of the antireceptor antibody, it was possible to successfully immunoprecipitate the biotinylated receptor. Furthermore, biotinylated receptors retained the ability to bind insulin and undergo insulin-stimulated autophosphorylation and internalization. The use of enzyme-labeled streptavidin enables the use of chemiluminescence techniques to detect the receptors, thus obviating the need to employ radioactivity. Just as the technique is useful to study cell surface insulin receptors, it can be adapted to investigate other cell surface receptors and proteins. PMID- 8404623 TI - Estrogen-regulated synthesis of neurotensin in neurosecretory cells of the hypothalamic arcuate nucleus in the female rat. AB - Neurotensin (NT) is implicated as a neurohormone in mammals, yet the peptide's neuroendocrine role remains to be determined. NT immunoreactivity has been observed in neurosecretory cells of the arcuate nucleus and paraventricular nucleus, and data suggest that NT release into hypophysial portal blood mediates a component of PRL secretion that is female specific and dependent on ovarian steroids. In the present study, in situ hybridization histochemistry and immunohistochemistry were used to investigate the regulation of NT gene expression in hypothalamic neurosecretory regions of adult rats. In ovariectomized females, estradiol induced expression of messenger RNA (mRNA) encoding NT and neuromedin N (NT/N mRNA) in the dorsomedial division of the arcuate nucleus. In contrast, estradiol did not appreciably alter NT/N mRNA expression in the ventrolateral division of the arcuate nucleus, where labeled cells were numerous, or in the paraventricular nucleus, where labeled cells were virtually absent. Estradiol also increased NT immunoreactivity in the external zone of the median eminence, confirming the neuroendocrine phenotype of NT cells in the dorsomedial division, as well as estrogen-regulated synthesis of NT in this system. In the dorsomedial division of cycling females, NT/N mRNA-expressing cells were significantly more numerous at proestrus than at diestrus, consistent with differences in plasma estradiol levels at these stages. In this same region, NT/N mRNA-expressing cells were significantly more numerous in proestrous females than in gonad-intact males. These results imply that estrogen-regulated release of NT at the median eminence subserves one or more sexually differentiated functions and are consistent with the involvement of tuberoinfundibular NT in estrogen-dependent secretion of PRL or GnRH on the afternoon of proestrus. PMID- 8404624 TI - Corticosteroid-binding globulin receptor of the rat hepatic membrane: solubilization, partial characterization, and the effect of steroids on binding. AB - The corticosteroid-binding globulin (CBG) receptor of rat hepatic membranes was solubilized using 1.5% Triton X-100. An assay for its activity was developed that was dependent upon the fact that the [125I] CBG-receptor complex adsorbs to hydroxylapatite, whereas [125I]CBG does not. Scatchard analysis of the soluble receptor at 37 C showed a single set of high affinity binding sites, with a Kd of 44 nM and a binding capacity of 7.3 pmol/mg protein. The association rate constant (k1) was 0.92 x 10(5) M-1 min-1 at 37 C, and the dissociation rate constant (k2) was 1.0 x 10(-3) min-1. Only unliganded CBG could bind to the receptor. Steroids that bound to CBG, e.g. corticosterone and cortisol, noncompetitively inhibited CBG's binding to the receptor. Steroids that did not bind to CBG, e.g, dexamethasone, were without effect on the interaction of CBG with its receptor. PMID- 8404625 TI - The phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate stimulates the dephosphorylation of mitochondrial ferredoxin in cultured chick kidney cells. AB - The tumor promoting phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA), presumably through activation of protein kinase C, decreases the production of 1 alpha, 25-dihydroxyvitamin D3 [1,25(OH)2D3] and increases that of 24R,25 dihydroxyvitamin D3 [24,25(OH)2D3] by primary cultures of chick kidney cells. We have previously shown that the regulation of the cellular output of 1,25(OH)2D3 and 24,25(OH)2D3 by PTH and 1,25(OH)2D3 can be quantitatively accounted for by altered hydroxylase activities within isolated mitochondria. In the present paper, we examined the effects of TPA and 1-oleoyl-2-acetyl-glycerol (OAG) on the state of mitochondrial protein phosphorylation and on 25-hydroxyvitamin D3 [25(OH)D3] metabolism. There was a good correlation between 25(OH)D3- 1 alpha- and 24-hydroxylase activities in mitochondria isolated from cells pretreated with either TPA or OAG and the pattern of 1- and 24-hydroxylation of 25(OH)D3. The most notable change in protein phosphorylation in the molecular mass range of 10 60 kilodaltons (kDa) was a dramatic decrease in the phosphorylation of a 12.5-kDa mitochondrial matrix protein after treatment of kidney cells with TPA or OAG. The amino acid composition of the 12.5-kDa protein was similar to bovine and human ferredoxins and it comigrated with bovine and human ferredoxins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The 12.5-kDa phosphoprotein was immunoprecipitated specifically by an antipeptide polyclonal antibody for chick ferredoxin. The dephosphorylation of ferredoxin in response to TPA was both rapid and transient, with the phosphate content of the 12.5-kDa protein reduced by 70% after a 5-min exposure and returning to control levels by 20 min. A similar transience was observed with regard to the rapid effects of TPA on 1 alpha hydroxylase activity, again showing maximal inhibition at 5 min. The results of our studies are consistent with the idea that ferredoxin phosphorylation plays a role in the regulation of steroid hydroxylation. PMID- 8404626 TI - Pregnancy lactogens in the rat conceptus and fetus: circulating levels, distribution of binding, and expression of receptor messenger ribonucleic acid. AB - To clarify the roles of the rat placental lactogens in embryogenesis and fetal development, we measured the concentrations of rat placental lactogen-II (rPL-II) in fetal rat serum and examined the distribution and expression of rPL-I- and rPL II-binding sites in rat uteroplacental and fetal tissues. The concentration of rPL-II in fetal rat serum on day 20 of gestation was 28.3 +/- 0.8 ng/ml (mean +/- SEM; n = 6), approximately 1/14th its concentration in maternal serum (398.3 +/- 45.3 ng/ml; n = 6). In the midgestational uterus and placenta, rat PL-I bound specifically to mesometrial decidua and to a capsular layer of stroma overlying the antimesometrial decidua. The binding of radiolabeled rPL-I to these tissues was inhibited by unlabeled rat PRL and human (h) GH, but not by rat GH, suggesting that the rPL-I-binding sites are lactogenic in nature. In the late gestational fetus, rat PL-II bound specifically to fetal adrenal, kidney, small intestine, liver, and pancreas; its binding, like that of rPL-I, was inhibited by rPRL, but not by rGH. rPL-II-binding sites in fetal adrenal were detected as early as day 16, whereas rPL-II-binding sites in fetal kidney and small intestine were not demonstrable until day 18. Lactogenic binding sites in fetal liver and pancreas did not appear until days 19-20. The relative amounts of specific binding of rPL-II to fetal tissues correlated positively with tissue levels of expression of the 4.2- and 1.8-kilobase PRL receptor mRNA transcripts. Radiolabeled hGH, which interacts with somatogenic receptors as well as lactogenic receptors, bound specifically to mesometrial decidua, fetal adrenal, kidney, small intestine, liver, and pancreas. In addition, radiolabeled hGH bound specifically, but with low intensity, to fetal brain. In mesometrial decidua and fetal adrenal, kidney, and small intestine, the binding of hGH was blocked by rPL II and rPRL, but not by rGH or ovine GH, suggesting the predominance of lactogenic receptors. In contrast, in fetal brain, the binding of hGH was inhibited by rGH, but not by rPL-II, suggesting that the fetal brain contains somatogenic receptors. The presence of rPL-I-binding sites in maternal decidua suggests a paracrine role for the hormone in decidual function at midgestation. The presence of rPL-II in fetal serum and the widespread distribution of rPL-II binding sites in fetal tissues indicate a role for rPL-II in fetal development. PMID- 8404627 TI - Role of growth hormone and insulin-like growth factor-I in mammary development. AB - Mammary glands from 3- to 4-week-old mice were incubated in whole organ culture to determine the effects of GH, PRL, and insulin-like growth factor-I (IGF-I) on lobulo-alveolar development and milk protein expression. Virgin mice were implanted with pellets of estrogen and progesterone (1:1000). After 9 days, abdominal no. 4 glands were removed and place on siliconized lens paper in Waymouths' medium supplemented with insulin (Ins), aldosterone, hydrocortisone, and epidermal growth factor. Concentrations of bovine GH, ovine GH, rat GH, or ovine PRL added to the medium varied from 0-1 micrograms/ml. IGF-I was added to replace either Ins or PRL up to 1 microgram/ml. When glands were incubated with Ins, aldosterone, hydrocortisone, and 250 ng/ml PRL, they exhibited lobulo alveolar development and expressed the milk protein beta-casein. When GH was substituted for PRL, little lobulo-alveolar development occurred, although beta casein mRNA was expressed at low levels. Either PRL or GH at 1 microgram/ml induced lobulo-alveolar development and beta-casein mRNA. Addition of epidermal growth factor to whole organ culture with GH or PRL (1 microgram/ml) was equally effective in stimulating lobulo-alveolar development. IGF-I did not substitute for PRL, GH, or insulin in tissue maintenance. It is clear that GH at high concentrations can act directly on mouse mammary tissue to induce both lobulo alveolar development and casein expression. PMID- 8404628 TI - Expression of vasopressin V1a and V2 receptor messenger ribonucleic acid in the liver and kidney of embryonic, developing, and adult rats. AB - The ontogenic expression of mRNAs encoding the V1a and V2 vasopressin receptors (V1aR and V2R) was visualized in liver and kidney of embryonic, developing, and adult rats using in situ hybridization histochemistry. Transcripts were detected at 16 and 19 days gestational age in kidney, with V1aR mRNA predominating in the developing cortex and V2R in the medulla. V1aR transcripts in 1-day-old kidneys were in vascular elements, in cells of developing medullary collecting ducts, and over mesangial cells of deep glomeruli, consistent with a role for the V1aR in regulating cellular growth. Expression of V1aR mRNA in the adult was found mainly in medullary vascular elements, arcuate and interlobular arteries, short segments of the cortical distal tubule, and transitional epithelium and smooth muscle of the pelvic wall and ureter. V2R mRNA, at 16 and 19 days gestational age, was in cells of developing medullary and cortical collecting ducts and, after birth, in cells of differentiating thick limbs of the loops of Henle, papillary surface epithelium, overlying macula densa, and short distal nephron segments. This distribution is in accord with the known role of V2 receptors in regulating water excretion. In contrast to kidney, liver did not express V2R mRNA and expressed V1aR transcripts only after birth. V1aR mRNA labeling was over cells bordering central veins on day 1 and surrounding central veins by day 5. A gradient was maximal on postnatal day 21, with V1aR mRNA most abundant in hepatocytes surrounding central veins and virtually absent in periportal hepatocytes. By day 60, most hepatocytes expressed V1aR transcripts, and the gradient was reduced. The ontogenic expression and receptor mRNA gradient are consistent with a role for hepatic V1a receptors in glucoregulation. These experiments confirm the presence of both V1a and V2 receptors in kidney and show that vasopressin receptor mRNA expression is developmentally regulated and tissue specific. PMID- 8404629 TI - Growth and cellular proliferation of ovine corpora lutea throughout the estrous cycle. AB - This study was conducted to determine the rates of growth and cellular proliferation of ovine corpora lutea (CL) throughout the estrous cycle. To determine the cellular labeling index (LI), ewes received an iv injection of bromodeoxyuridine (BrdU) 1 h before death on days 2, 4, 8, 12, or 15 (day 0 = estrus; n = 6-12 ewes/day). At death, CL were weighed, and samples of each were fixed in Carnoy's solution or frozen until analyzed for DNA, protein, and progesterone contents. Nuclear incorporation of BrdU was determined in paraffin embedded tissue sections by using a primary antibody against BrdU and a fluorescent (fluorescein isothiocyanate-labeled) secondary antibody, and sections were counterstained with propidium iodide (a nuclear stain). The labeling index (BrdU-labeled nuclei as a proportion of propidium iodide-labeled nuclei) of each CL was determined by using dual channel interactive laser cytometry and image analysis. Moreover, BrdU and 3 beta-hydroxysteroid dehydrogenase (a marker for steroidogenic cells) or BrdU and factor VIII (a marker for endothelial cells) were immunolocalized in tissue sections by using double immunohistochemical or dual immunofluorescent staining, respectively. Results demonstrated that cellular proliferation was greatest (LI, 34.1 +/- 2.1%) on day 2 and decreased (P < 0.01) through day 15 (LI, 0.7 +/- 0.1%) of the estrous cycle. The results of the immunohistochemical studies provide evidence that both parenchymal (steroidogenic) and nonparenchymal (e.g. endothelial, fibroblastic) luteal cells proliferated throughout the ovine estrous cycle. Conversely, from days 2-12 of the estrous cycle, fresh weight and DNA content of CL increased linearly (P < 0.01; 8- and 10-fold, respectively), then decreased (P < 0.02) from days 12-15. Ratios of protein/DNA on days 2, 4, and 8 were similar and were greater (P < 0.02) than those on days 12 and 15, which also were similar. These data demonstrate that growth of the ovine CL is extremely rapid, linear from days 2 12, and primarily due to hyperplasia. In addition, the high rate of cellular proliferation is associated primarily with nonsteroidogenic cells, a large proportion of which appear to be endothelial cells. Data such as these will enable us to determine the factors that are important in regulating luteal growth and development in normal and pathological conditions. PMID- 8404630 TI - Platelet-derived growth factor effects on purified testicular peritubular myoid cells: binding, cytosolic Ca2+ increase, mitogenic activity, and extracellular matrix production enhancement. AB - The response of purified rat testicular peritubular myoid cells (PMC) to platelet derived growth factor (PDGF) was studied. Freshly isolated PMC were devoid of measurable amounts of PDGF-binding sites. However, after 1 day in culture in serum-free conditions, specific high affinity receptors were detected. The estimated binding sites per cell revealed that PMC express more receptors for PDGF-BB, followed by PDGF-AB and PDGF-AA. PDGF treatment of cultured PMC increased the cytosolic Ca2+ concentration, showing a rank order of potencies with PDGF-BB > PDGF-AB > PDGF-AA. PMC proliferation, as measured by direct cell counting, was also stimulated by all three PDGF isoforms, with the same order of potencies observed for the increase in intracellular Ca2+. This effect was inhibited by antibodies to PDGF. Moreover, PDGF treatment increased the release of type IV collagen and fibronectin, and induced the release of type V collagen and laminin. These results demonstrate that testicular PMC are induced to express functionally active PDGF receptors in response to cell culturing. These data suggest that PMC may be a target for PDGF and that PDGF-mediated effects in vivo are dependent on factors regulating the expression of the receptors. The role that PDGF may play in normal and pathological testicular processes is discussed. PMID- 8404631 TI - Evidence that neuropeptide Y could represent a neuroendocrine inhibitor of sexual maturation in unfavorable metabolic conditions in the rat. AB - Neuropeptide Y (NPY) is known to be involved in the central regulation of appetite, sexual behavior and reproductive function. Whereas central administration of NPY strongly stimulates feeding, diet restriction produces overexpression of NPY in the arcuate and paraventricular nuclei that might reflect behavioral adaptations to shortage of food. The role of NPY for the regulation of sexual function is still controversial. Whereas NPY is stimulatory during proestrus in the rat, acute administration of NPY is inhibitory in castrated animals and we have shown that chronic administration of NPY inhibits both the gonadotropic and somatotropic axis in adult female rats. In order to further analyse the role of NPY during sexual maturation, a model of delayed sexual maturation imposed by food restriction and return to ad-libitum feeding was used. Young female rats were restricted to 7-8 g food daily starting at 24 days of life (d). This restriction completely prevented sexual maturation. At 50 d, ICV cannulas were placed and at 60 d, Alzet minipumps either delivering NPY (18 micrograms/day) or vehicle into the ICV cannula were implanted dorsally. At 61 d, rats were switched to ad-libitum feeding, a change that produced vaginal opening within 4 days in all vehicle-treated rats. In the rats receiving NPY, significantly increased food intake and weight gain were observed but only one out of the 9 rats studied experienced vaginal opening at 66 d, the other 8 animals remaining sexually immature at 67 d at sacrifice. Sexual immaturity of NPY-treated rats was further confirmed by decreased ovarian weight and reduced number of pituitary GnRH receptors. Plasma IGF-I levels were markedly reduced in NPY-treated rats. Since food restriction has been shown both to increase hypothalamic NPY and to reduce or inhibit sexual function, these data bring evidence for the first time that NPY could be involved in the inhibition of sexual maturation imposed by food restriction, since maintenance of elevated NPY levels in the hypothalamus did prolong this state of sexual immaturity despite restoration of normal food intake. PMID- 8404632 TI - Nitric oxide inhibits uterine contractility during pregnancy but not during delivery. AB - Nitric Oxide mediates various biological phenomena, including vascular smooth muscle relaxation. In the present study, we sought to determine if an L-arginine nitric oxide-relaxation system is present in the uterus and if it modulates contractility during pregnancy. The substrate and a donor of nitric oxide and nitric oxide gas caused substantial relaxation of the spontaneous contractility of tissues from the rat uterus in vitro during pregnancy. Inhibitors of nitric oxide synthase and soluble guanylate cyclase reversed the relaxation effects of L arginine. Nitric oxide was produced by the uterus in organ culture. Relaxation effects of L-arginine on the pregnant rat uterus were diminished at the time of spontaneous labor and postpartum. Nitric oxide production was also substantially reduced during labor. These results show that an L-arginine-nitric oxide relaxation system is present in the uterus and it inhibits contractility during pregnancy but not during labor. PMID- 8404633 TI - Localization of C-type natriuretic peptide mRNA in rat hypothalamus. AB - Central or peripheral administration of C-type natriuretic peptide (CNP) affects numerous neuroendocrine systems, including the hypothalamo-pituitary-gonadal, hypothalamo-pituitary-adrenocortical and hypothalamo-neurohypophysial axes. The present report characterizes the distribution of CNP mRNA in hypothalamus, providing the first definition of CNP-containing neuroendocrine circuits. In situ hybridization histochemical analysis revealed high expression of CNP mRNA in the anteroventral periventricular nucleus (AVPv) and in hypothalamic arcuate nucleus (ARC). Hybridization signals of significantly lower intensity were seen in the medial, median and periventricular preoptic area, the supraoptic, dorsomedial, ventral premammillary and lateral mammillary nuclei and in the posterior hypothalamic area. A few scattered CNP mRNA containing cells were visualized in the medial parvocellular paraventricular nucleus, posterior magnocellular paraventricular nucleus and lateral hypothalamic area. In the AVPv and ARC the pattern of CNP mRNA distribution paralleled that of ANP mRNA. The results indicate a distribution of CNP mRNA associated with key neuroendocrine systems, and underscores the potential importance of this novel natriuretic peptide in neuroendocrine regulation. PMID- 8404634 TI - Cloning and functional expression of the human glucagon-like peptide-1 (GLP-1) receptor. AB - Truncated forms of glucagon-like peptide-1 are the most potent endogenous stimuli of insulin secretion and have powerful antidiabetogenic effects. To determine the structure and coupling mechanisms of the human GLP-1 receptor we have isolated two pancreatic islet cDNAs, encoding the 463 amino acid receptor and differing mainly in their 3' untranslated regions. The deduced amino acid sequence is 90% homologous with the rat GLP-1 receptor. Northern blot analysis shows expression of a single 2.7 kb transcript in pancreatic tissue. When expressed in COS-7 cells the recombinant receptor conferred specific, high affinity GLP-1(7-37) binding. GLP-1(7-37) increased intracellular cAMP in a concentration dependent manner and caused an increase in the free cytosolic calcium ([Ca2+]i) from an intracellular pool, characteristic of phospholipase C (PLC) activation. Thus, like the structurally related glucagon and parathyroid hormone receptors, the human GLP-1 receptor can activate multiple intracellular signaling pathways including adenylyl cyclase and PLC. Knowledge of the GLP-1 receptor structure will facilitate the development of receptor agonists and elucidation of the important role of GLP-1 in normal physiology and disease states. PMID- 8404635 TI - Dynamic regulation of gonadotropin-releasing hormone receptor mRNA levels in the anterior pituitary gland during the rat estrous cycle. AB - The number of gonadotropin-releasing hormone (GnRH) receptors on pituitary gonadotropes varies substantially during the rat estrous cycle and may modulate pituitary responsiveness to GnRH. The present studies were undertaken to determine to what extent these changes in GnRH receptor number reflect a change in GnRH receptor mRNA expression in the anterior pituitary gland. Using quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), pituitary GnRH receptor mRNA levels were measured at various timepoints throughout the rat estrous cycle. There was a three-fold increase in GnRH receptor mRNA levels on the afternoon of proestrus (PRO) when compared to levels observed on the morning of metestrus (MET). This rise preceded the onset of the LH surge by 6h (1200h). GnRH receptor mRNA levels remained elevated through 2100h PRO, after which they dropped dramatically, and by 2400h PRO were not significantly different from levels observed at 0900h MET. A two-fold increase in GnRH receptor mRNA expression was also observed during the early stages of the estrous cycle (0900h to 1800h MET), and this increase was sustained until 1800h on diestrus, at which time mRNA levels decreased to levels observed at 0900h MET. These results demonstrate that pituitary GnRH receptor mRNAs are dynamically regulated during the rat estrous cycle, with receptor mRNA expression being greatest on the afternoon of PRO, the time of the estrous cycle at which gonadotropes are most sensitive to GnRH stimulation. PMID- 8404636 TI - Finasteride blocks progesterone synthesis in MA-10 Leydig tumor cells. AB - The 5 alpha-reductase inhibitor, finasteride, inhibits progesterone synthesis in the MA-10 Leydig tumor cells. Inhibition is dose-dependent with half maximal inhibition occurring at 10 ng/ml, a concentration significantly less than serum concentrations detected in finasteride-treated patients. Experiments to localize the site of inhibition by this compound revealed that the 3 beta-hydroxysteroid dehydrogenase delta 5-->delta 4 isomerase enzyme was not blocked by finasteride, but that cholesterol side-chain cleavage was inhibited. Thus, both dibutyryl-cAMP stimulated and 22-hydroxycholesterol-stimulated steroidogenesis were inhibited by finasteride. This effect of finasteride to block cholesterol side-chain cleavage may be species-specific. Inhibition is readily detected in the mouse-derived MA 10 cells; however, human granulosa cell steroidogenesis is finasteride insensitive while rat Leydig cell steroidogenesis is only minimally effected by finasteride. PMID- 8404637 TI - What do we know about the expression of proopiomelanocortin transcripts and related peptides in lymphoid tissue? PMID- 8404638 TI - Analysis of proopiomelanocortin (POMC) messenger ribonucleic acid and POMC derived peptides in human peripheral blood mononuclear cells: no evidence for a lymphocyte-derived POMC system. AB - A number of recent studies suggest that cells of the immune system, e.g. peripheral blood mononuclear cells (PBMC), can synthesize and process POMC and secrete POMC-derived peptides, such as ACTH and endorphins, upon immune and hormonal challenges. From this, it has been proposed that POMC-derived peptides originating from lymphoid cells can function as hormones, for instance in a lymphoid-adrenal axis. In view of the important physiological implications of this proposal, the present study was designed to investigate the expression of the POMC gene in human PBMC and the production by these cells of alpha-, beta-, and gamma-endorphins (alpha E, beta E, and gamma E) peptides that are established end products of the posttranslational processing of POMC. PBMC of individual donors were used uncultured (fresh cells) or cultured for 24 and 48 h in the presence and absence of Concanavalin-A (Con-A), bacterial lipopolysaccharide, phytohemagglutinin, or CRH, and vasopressin, conditions that reportedly stimulate POMC activity in those cells, to investigate the presence of POMC transcripts by analysis of total RNA with Northern blotting and the reverse transcriptase polymerase chain reaction (RT-PCR). Large scale preparations containing over 10(9) cells (fresh, cultured with and without Con-A) originating from several donors were examined for the presence of POMC transcripts by analysis of poly(A)+ RNA on Northern blots and for the presence of alpha E, beta E, and gamma E by gel filtration over Sephadex G-75 and reverse phase HPLC, followed by assay of the fractions in four endorphin RIA systems with different specificities. On the Northern blots of total RNA, no POMC transcripts were detectable. In poly(A)+ RNA preparations, no full-length POMC mRNA was found, and it was estimated that the concentration of POMC mRNA, if present, was below approximately 0.005 transcript/cell in Con-A-stimulated cells and still lower in unstimulated cells. In accord with literature data, an 800- to 900-nucleotide POMC transcript was detected in cultured PBMC, and the levels of this transcript were stimulated by Con-A. In all samples analyzed with RT-PCR, a transcript spanning most of exons 2 and 3 was detectable only on Southern blots of the RT-PCR product, but not on agarose gels stained with ethidium bromide. Chromatographic analysis of endorphin immunoreactivities in cell extracts revealed no qualitative differences between the immunoreactive profiles of fresh PBMC or PBMC cultured with or without Con A.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404639 TI - Protooncogene junB as a target for activin actions. AB - Activin, a member of the transforming growth factor-beta family of peptides, is implicated in the regulation of cell growth and differentiation in a variety of biological systems. We have sought to identify immediate early genes whose altered expression may provide a common nuclear event involved in activin regulated phenotypic changes in many cell types. In both human K562 myelogenous leukemia and rat PC12 pheochromocytoma cells, activin treatment caused transient transcription-dependent and protein synthesis-independent increases of junB messenger RNA (mRNA) within 1 h, whereas neither c-jun nor c-fos mRNA were inducible. In K562 cells, this selective junB mRNA induction was synergistically augmented by treatment with 12-O-tetradecanoyl phorbol-13-acetate but not affected by forskolin. Furthermore, in PC12 cells, the up-regulation of junB mRNA by activin was observable even after high-dose treatment with 12-O-tetradecanoyl phorbol-13-acetate for 48 h, indicating that junB mRNA expression by activin is independent of both A- and C-kinases. Our report suggests that induction of this ubiquitous gene product may be a critical event shared by a set of activin responsive tissues. PMID- 8404640 TI - Diurnal differences in basal and acute stress levels of type I and type II adrenal steroid receptor activation in neural and immune tissues. AB - To examine diurnal differences in the proportions of receptors that were occupied and activated by basal and stress levels of corticosterone, we measured available type I (mineralocorticoid) and type II (glucocorticoid) adrenal steroid receptor levels in brain, pituitary, and immune tissues of unstressed and acutely stressed rats at the times of day when basal corticosterone secretion was at its trough [morning (AM)] and peak [evening (PM)]. In general, the estimated adrenal steroid receptor activation was greater in brain than in pituitary or immune tissue, and within a particular tissue, there was a greater degree of estimated activation of the adrenal steroid high affinity type I receptor than of the type II receptor. There was a greater activation of brain type II receptors by basal corticosterone in the PM (30-35%) than the AM (5-15%). As acute stress produced similar levels of receptor activation at both times of day (45-50%), the net change in type II receptor activation in the brain after acute stress was much smaller in the PM than in the AM. This suggests that there may be diurnal differences in the role of type II receptors in corticosterone negative feedback on the hypothalamic pituitary-adrenal axis. In immune tissues, type II receptor activation by acute stress was especially heterogeneous, depending on both the immune compartment and the time of day, suggesting that these are important factors contributing to a differential impact of corticosterone on immune responses during acute stress. Taken together our results suggest that the tonic and phasic influences of corticosterone on target tissue responses very not only with the diurnal and stress secretion patterns of corticosterone, but also with target tissue factors, such as type I and type II receptor expression and hormone bioavailability. All of these factors contribute to considerable selectivity of action for the systemic hormone corticosterone. PMID- 8404641 TI - A study of the 3,5,3'-triiodothyronine sulfation activity in the adult and the fetal rat. AB - We have employed a new in vitro assay for study of the T3 sulfation activity in rat tissues. The assay measures by RIA the generation of T3 sulfate (T3S) during incubation of T3 with cytosol of rat tissues as the source of phenol sulfotransferase(s) and 3-phosphoadenosine-5'-phosphosulfate as the sulfate donor. The conversion of T3 to T3S proceeded rapidly for 30 min at 37 C, and the optimal pH of the reaction was 8.0. Heating the cytosol at 44 C for 15 min decreased T3S production to 63% of its value at 37 C. T3 sulfation activity was plentiful in rat liver, brain, and kidney, but little activity was demonstrable in other tissues. The Km and maximum velocity of the hepatic conversion of T3 to T3S were 114 microM and 159 pmol/mg protein.h, respectively. There was a marked inhibition of the conversion of T3 to T3S with salicylamide, 3' monoiodothyronine, thyronine, and rT3; the IC50 of these inhibitors approximated 15, less than 0.1, 9.5, and 43 microM, respectively. On day 17 of gestation, the T3 to T3S conversion activity was more abundant in fetal skin than in other fetal tissues. However, the activity decreased in fetal skin while it increased in fetal liver, kidney, and brain nearer to term on day 20. Placenta demonstrated lower T3 to T3S conversion activity than several fetal or maternal tissues. There was no effect of hypothyroidism or hyperthyroidism on T3 sulfation activity. We conclude that T3 sulfation activity in the rat is 1) most abundant in liver, kidney, and brain tissues of the adult; 2) inhibited more avidly by 3' monoiodothyronine than other thyronines; 3) very abundant in fetal skin early in gestation; and 4) little affected by the thyroidal status of the animal. PMID- 8404642 TI - The effect of cholecystokinin on intracellular Ca2+, membrane-associated protein kinase-C activity, and progesterone production in chicken granulosa cells. AB - Nerve fibers immunoreactive for cholecystokinin (CCK) have been observed in the rat ovary, but the function of this gut peptide in the ovary is not known. These studies were designed to investigate the effects of the CCK C-terminal octapeptide (CCK-8) on the intracellular calcium ion concentration ([Ca2+]i), protein kinase-C (PKC) activity, and progesterone secretion in granulosa cells obtained from the two largest preovulatory follicles (F1 and F2) of hens. [Ca2+]i was measured in cells loaded with the Ca(2+)-responsive fluorescent dye fura-2. The resting [Ca2+]i in these cells was 96 +/- 5 nM. There was a rapid (i.e. within 5-10 sec) 2- to 4-fold increase in [Ca2+]i in 70% of the cells examined after the addition of 10(-7) M CCK-8. The CCK-8-triggered [Ca2+]i transient was not affected by incubating the cells in Ca(2+)-free medium containing 2 mM EGTA or by pretreating the cells with a Ca2+ channel blocker, such as La3+ (1 mM) or D600 (100 microM). The CCK-8-triggered [Ca2+]i surge was abolished by pretreating the cells with the inhibitor of inositol phospholipid hydrolysis, neomycin (1.5 mM), the CCK antagonists proglumide (1 mM) and benzotript (1 mM), or pertussis toxin (50 ng/ml for 12 h). Incubating granulosa cells with CCK-8 (2 x 10(-7) M) for 10 min stimulated a 1.60 +/- 0.04-fold increase in membrane-associated PKC activity over control levels. In 3-h incubations, CCK-8 (10(-6)-10(-8) M) did not affect basal or LH (20 or 100 ng/ml-stimulated progesterone production. These studies demonstrate that CCK-8 causes a transient increase in chicken granulosa cell [Ca2+]i through the release of Ca2+ from intracellular stores and activates membrane-associated PKC activity, but does not affect progesterone production. These results suggest the presence of G-protein-coupled phospholipase-C activating CCK receptors on the surface of these cells. PMID- 8404643 TI - Glucocorticoids inhibit soluble phospholipase C activity and cytosolic guanine nucleotide regulatory protein-alpha i immunoreactivity in spleen. AB - Glucocorticoids have a wide range of effects in mammalian tissues. In this study we investigated the hypothesis that some of the long-term effects of glucocorticoids on immune cell function may occur by regulating phospholipase C (PLC) signal transduction systems that are known to play a role in immune cell activation. PLC activity was measured in vitro with [3H]phosphatidylinositol and [3H]phosphatidylinositol 4,5-bisphosphate as substrates. Although the guanine nucleotide regulatory protein, Gi, is a membrane-associated protein, our unpublished observations show significant Gi levels in the cytosol of several tissues, including the spleen, where the highest levels were detected. We measured cytosolic Gi alpha immunoreactivity after hormone treatment to establish the relationship between the regulation of cytosolic PLC activity and cytosolic Gi alpha levels. GTP and its nonhydrolyzable analogs have been shown in some instances to regulate soluble PLC activity. In vivo administration of dexamethasone (DEX; 5 mg/ml, sc) to rats for 24 h reduced soluble PLC activity from spleen by 25-50%. In the same tissue, cytosolic Gi alpha immunoreactivity was decreased by 60-70%. The time dependency and receptor specificity of the glucocorticoid effects observed in vivo were investigated further in isolated splenocytes. Treatment of intact splenocytes with DEX (10(-8) and 10(-7) M) for 48 h inhibited calcium-stimulated cytosolic PLC activity by 80-90%; cytosolic Gi alpha immunoreactivity was also reduced by about 90%. In a time course experiment with DEX (10(-7) M) in splenocytes, significant effects were apparent by 12 h after steroid treatment and were maximal by 48 h. When splenocytes were coincubated with DEX (10(-8) M) and the glucocorticoid receptor antagonist RU 486 (10(-7) M), the effects of DEX on soluble PLC activity and cytosolic Gi alpha immunoreactivity were both blocked, suggesting that the effects were mediated by DEX activation of the classical intracellular glucocorticoid receptor. The effects of glucocorticoids reported here may represent one way by which these hormones act to modulate immune cell function. PMID- 8404644 TI - Isolation and characterization of a complementary deoxyribonucleic acid insert encoding bovine aromatase cytochrome P450. AB - Aromatase, an enzyme complex comprised of aromatase cytochrome P450 (P450arom; the product of the CYP19 gene) and the flavoprotein NADPH-cytochrome P450 reductase, catalyzes the conversion of androgens to estrogens. Three cDNA inserts encoding P450arom were isolated from a bovine placental cDNA library. These inserts were sequenced and found to correspond closely to human P450arom sequence from the internal EcoRI restriction site (exon III) through the termination codon (exon X) into the 3'-untranslated region. The rapid amplification of cDNA ends technique was used to generate the rest of the cDNA 5' of the internal EcoRI site, using mRNA obtained from bovine granulosa cells as a template. This insert was sequenced, and when aligned with the other inserts, an open reading frame was found which was predicted to encode a protein of 503 amino acid residues. The deduced polypeptide shares 84% identity with human P450arom and 79%, 76%, 71%, and 57% identity with mouse, rat, chicken, and trout P450arom, respectively. A full-length open reading frame was generated using the polymerase chain reaction and mRNA obtained from bovine granulosa cells as template. After this insert was ligated into the pCMV5 expression vector, it was transfected into COS-1 monkey kidney tumor cells. We were able to demonstrate aromatase activity by assaying the incorporation of tritium into [3H] water from [1 beta-3H]androstenedione. Northern analysis revealed a single transcript of approximately 6 kilobases in poly(A)+ RNA obtained from bovine placental tissue and granulosa cells. This indicated for the first time a correspondence between the pattern of estrogen biosynthesis throughout the bovine ovarian cycle and the levels of transcripts encoding P450arom. In addition, weak hybridization was noted to transcripts of the expected size, namely 3.4 and 2.9 kilobases, in poly(A)+ RNA obtained from human placental tissue. The large size of the bovine transcript is due to a long 3'-untranslated region, because, based on the rapid amplification of cDNA ends technique, there appeared to be approximately 150 basepairs 5' of the start site of translation, and we were never able to find a polyadenylation site, even in one clone that went well past the corresponding polyadenylation site in human P450arom. PMID- 8404645 TI - Differential expression of gamma-aminobutyric acid receptors in immortalized luteinizing hormone-releasing hormone neurons. AB - gamma-Aminobutyric acid (GABA) has been shown both to stimulate and inhibit LH secretion in vivo. GABA apparently exerts these effects at the hypothalamic level by regulating the release of LHRH. In this study, we have investigated the effect of GABAergic agents on LHRH secretion from an immortalized hypothalamic neuronal cell line (GT1-7). LHRH secretion was stimulated in a dose-dependent manner with increasing concentrations of GABA. This effect was mimicked by the GABAA receptor agonist, muscimol, and was blocked by the selective antagonist, bicuculline. The stimulatory effect of muscimol on LHRH secretion was synergistic with low concentrations of [K+]. By comparison, neither activation of the GABAB receptors with baclofen nor blockade with phaclofen influenced basal LHRH secretion. Baclofen, however, did depress [K+]-induced LHRH release. Binding studies confirmed the presence of GABAA and GABAB receptors on GT1-7 cells. In addition, Northern blots with probes to the GABAA receptor alpha 1, beta 3, and gamma 2L subunits revealed that only the beta 3 messenger RNA (mRNA) was expressed in the GT1-7 cells. These data provide the first demonstration that immortalized LHRH neurons are directly responsive to GABAergic agents. To the extent that these immortalized neurons may resemble those in vivo, our results suggest that GABAergic agents may play a dual role in reproductive physiology by exerting both stimulatory and inhibitory control over LHRH release. PMID- 8404646 TI - Retinoic acid differentially regulates expression of surfactant-associated proteins in human fetal lung. AB - Retinoic acid is known to play an essential role in maintaining the differentiation of a wide variety of epithelial cell types. However, its effects on the differentiation of lung alveolar epithelium have not been described. In the present study, we examined the effects of retinoic acid on the differentiation of human fetal lung tissue maintained in vitro. Human fetal lung explants were cultured in serum-free medium for 6 days in the absence or presence of all-trans retinoic acid at concentrations from 0.3 nM to 3 microM. Explant content of the surfactant-associated protein SP-A was measured using a specific enzyme-linked immunosorbent assay. Retinoic acid reduced SP-A protein levels in a concentration-dependent manner [analysis of variance (ANOVA), P < 0.01]. To evaluate possible cytotoxic effects of retinoic acid, culture media were assayed for lactate dehydrogenase (LDH), a cytoplasmic enzyme. LDH levels in media from retinoic acid-treated explants were not significantly different than LDH levels in media from control explants, indicating that retinoic acid is not cytotoxic in human fetal lung explants. Changes in messenger RNA (mRNA) levels for surfactant associated proteins SP-A, SP-B, and SP-C were measured by Northern blot analysis. Retinoic acid reduced SP-A mRNA levels in a concentration-dependent manner (ANOVA, P < 0.02) and reduced SP-C mRNA levels at 3 microM. In contrast, retinoic acid increased SP-B mRNA levels in a concentration-dependent manner (ANOVA, P < 0.03). Morphometric analysis showed that retinoic acid decreased epithelial volume density in the explants by approximately 17% and increased connective tissue volume density by approximately 20% when compared to dimethyl sulfoxide vehicle controls. These data indicate that retinoic acid regulates type II cell surfactant protein gene expression in human fetal lung tissue. PMID- 8404647 TI - Cloning and increased expression with fructose feeding of rat jejunal GLUT5. AB - We have isolated a clone from the rat jejunum cDNA library using a fragment of human GLUT5 cDNA as a probe. The coding region of this clone shares 80% nucleotide and 81% amino acid identity with human GLUT5 and is thus termed rat GLUT5 cDNA. Rat GLUT5 mRNA exhibited a tissue distribution very similar to that of human GLUT5, with the highest levels in the jejunum, but was not detected in fat, muscle, or testis on Northern blot analysis. The antipeptide antibody raised against the C-terminal domain of rat GLUT5 protein specifically recognized a rat jejunal protein with an apparent mol wt of 60,000 on immunoblots. The amount of GLUT5 mRNA and protein in the jejuni of rats fed a fructose-enriched diet (50%, wt/wt) for 3 days were increased 2.5- and 6-fold, respectively, compared to those of rats fed standard rat chow, whereas those of rats fed a starch-enriched diet (50%, wt/wt) were not altered. Similarly, GLUT5 mRNA and protein in the jejunum were increased 5- and 8-fold, respectively, after 15 days of fructose feeding. Thus, an increase in fructose absorption up-regulates GLUT5 expression in the jejunum. These results are consistent with the notion that GLUT5 plays a major role in fructose absorption in the small intestine. PMID- 8404648 TI - Influence of hypercholesterolemia on adrenal steroid metabolism and electrolyte balance in spontaneously hypertensive rats. AB - Hypercholesterolemia and hypertension are frequently associated risk factors for cardiovascular diseases. The interactions between hypercholesterolemia and the regulatory mechanisms of blood pressure are poorly understood. In this study we investigated the effects of hypercholesterolemia on salt metabolism and its hormonal control mechanisms in spontaneously hypertensive rats (SHR). Six-week old SHR were randomly assigned to either a high (1%) cholesterol diet or a matched regular diet for 6 weeks, followed by a 2-week dietary washout. A group of normotensive Wistar-Kyoto rats received the high cholesterol diet and was used as a control. Plasma cholesterol increased significantly (P < 0.001) in both cholesterol-fed SHR and Wistar-Kyoto rats. Blood pressure was unaffected by 6 weeks of a high cholesterol diet. Hypercholesterolemia caused a significant increase in aldosterone (by analysis of variance: F = 8.40; P < 0.01) associated with a significant decrease in corticosterone (F = 4.64; P < 0.05) in the SHR, but not in the normotensive rats. In addition, in the cholesterol-fed SHR, urinary sodium excretion was reduced (P < 0.01), and the urinary potassium/sodium ratio was increased (P < 0.01) compared to those in the remaining groups of rats. The hormonal and urinary differences between the hypertensive subgroups were not detectable after withdrawal of cholesterol. These results demonstrate that diet induced hypercholesterolemia specifically promotes reversible mineralocorticoid accumulation and sodium retention in SHR. PMID- 8404649 TI - The dissociation of tumor-induced weight loss from hypoglycemia in a transplantable pluripotent rat islet tumor results in the segregation of stable alpha- and beta-cell tumor phenotypes. AB - We previously established pluripotent transformed rat islet cell lines, MSL cells, of which certain clones have been used to study processes of islet beta cell maturation, including the transcriptional activation of the insulin gene induced by in vivo passage. Thus, successive sc transplantation in NEDH rats resulted in stable hypoglycemic insulinoma tumor lines, such as MSL-G2-IN. Occasionally, hypoglycemia as well as severe weight loss were observed in the early tumor passages of MSL-G and the subclone, NHI-5B, which carry the transfected neomycin and human insulin genes as unique clonal markers. By selective transplantation, it was possible to segregate stable anorectic normoglycemic tumor lines, MSL-G-AN and NHI-5B-AN, from both clones. These tumors cause an abrupt onset of anorexia when they reach a size of 400-500 mg (< 0.3% of total body weight), and the observed weight loss parallels that of starved rats until death results from cachexia. After tumor resection, animals immediately resume normal feeding behavior. Comparative studies of hormone release and mRNA content in anorectic lines, MSL-G-AN and NHI-5B-AN, vs. those in the insulinoma line, MSL-G2-IN, revealed selective glucagon gene expression in both of the anorectic tumors, whereas insulin and islet amyloid polypeptide gene expression were confined to the insulinoma. Both tumor phenotypes produced cholecystokinin and gastrin in variable small amounts, making it unlikely that these hormones contribute to the anorectic phenotype. Tumor necrosis factor (cachectin) was not produced by any of the tumors. Proglucagon was processed as in the fetal islet to products representative of both pancreatic alpha-cell and intestinal L-cell phenotypes, with glucagon and Glp-1 (7-36)amide as the major extractable products. In contrast to the administration of cholecystokinin, neither glucagon, Glp-1 (7-36)amide, nor their combination, affected feeding behavior in fasted mice, suggesting the presence of a hitherto unidentified anorectic substance released from the glucagonoma. We conclude 1) that glucagonomas and insulinomas can be derived from a common clonal origin of pluripotent MSL cells, thus supporting the existence of a cell lineage relationship between islet alpha- and beta-cell during ontogeny; and 2) that our glucagonomas release an anorexigenic substance(s) of unknown nature that causes a severe weight loss comparable to that reported in animals carrying tumor necrosis factor-producing experimental tumors. PMID- 8404650 TI - Expression of insulin-like growth factor-I (IGF-I) in the rat fallopian tube: possible autocrine and paracrine action of fallopian tube-derived IGF-I on the fallopian tube and on the preimplantation embryo. AB - Recent studies have indicated that growth factors such as insulin-like growth factors (IGFs) increase the growth rate of cultured preimplantation embryos. We therefore hypothesized that the fallopian tube may produce IGFs which in turn participate in the regulation of preimplantation embryo development in vivo. In the present study we examined the expression of IGF-I in the fallopian tube. We demonstrated that IGF-I transcripts (7.0, 1.7, and 1.2-0.8 kilobases) were abundant in the fallopian tube. Immunoreactive IGF-I was most abundant in the epithelial cells in the fallopian tube, and IGF-I messenger RNA (mRNA) was detected in the luminal region of the fallopian tube. A solution hybridization assay was used to examine the regulation of IGF-I mRNA. The abundance of IGF-I transcripts changed markedly during the 4-day estrous cycle with the highest levels on the day of proestrus. The increase in IGF-I mRNA between the day of diestrus II and the day of proestrus was 4-fold (P < 0.01). The pattern of IGF-I mRNA expression in the fallopian tube resembled the pattern of ovarian estrogen production during the estrous cycle. The level of IGF-I mRNA decreased after hypophysectomy. The expression of IGF-I mRNA in the fallopian tube was dose dependently regulated by estradiol, and a single sc injection of estradiol [5 micrograms/100 g body wt (BW)] increased the IGF-I mRNA in a time-dependent manner with a significant increase after 3 h (P < 0.01). The lowest estradiol dose tested (0.1 microgram/100 g BW) increased the expression after 6 h, whereas progesterone (5 micrograms/100 g BW) was ineffective. The presence of embryos in the fallopian tube did not statistically significantly influence the abundance of IGF-I transcripts as measured with a solution hybridization assay on RNA extracted from whole fallopian tubes. In order to determine possible targets for fallopian tube-derived IGF-I we examined the expression of IGF-I receptor mRNA. Northern blot analysis revealed that an 11-kilobase IGF-I receptor transcript was expressed in the fallopian tube. Using a reverse transcriptase-polymerase chain reaction, IGF-I receptor mRNA was also detected in the eight-cell but not two cell preimplantation embryo. The present study demonstrates that IGF-I is produced in the fallopian tube and its expression is regulated by estradiol. Both the fallopian tube and the eight-cell preimplantation embryo express IGF-I receptors and are therefore potential target tissues. PMID- 8404651 TI - Evidence for a role for the cyclic adenosine 3',5'-monophosphate/protein kinase-A pathway in regulation of the gonadotropin subunit messenger ribonucleic acids. AB - cAMP regulation of gonadotropin secretion and subunit mRNA levels was studied in pituitary cells perifused with pulses of GnRH. Pituitary cells from 7-week-old male rats castrated at 5 weeks of age were stimulated hourly for 9-24 h with 1 min pulses of GnRH, the adenylate cyclase activator forskolin, the cell-permeable cAMP analog 8-bromo-cAMP (8Br-cAMP), or control medium. Cells were also treated with the nonsteroidal antiinflammatory drug flufenamic acid, which reduces pituitary cAMP levels. During perifusion, the effluent was collected in 10-min fractions for FSH and LH assay. At the completion of perifusion, total RNA was extracted, and gonadotropin subunit mRNA levels were quantitated by Northern analysis. Continuous administration of flufenamic acid gradually reduced the amplitude of GnRH-stimulated FSH and LH pulses to nadir values of 40 +/- 4.7% and 62 +/- 12% of the control value, respectively. Flufenamic acid decreased (P < 0.05) FSH beta and alpha-subunit mRNA levels and blocked the effect of GnRH to lengthen LH beta mRNA. Pulses of forskolin or 8Br-cAMP released LH and FSH, and continuous forskolin or 8Br-cAMP potentiated the gonadotropin stimulatory effect of GnRH. Forskolin or 8Br-cAMP increased (P < 0.05) FSH beta mRNA and alpha subunit mRNA levels when administered in pulses, but not when administered continuously, and lengthened LH beta mRNA. The Nal-Glu GnRH antagonist blocked the effects of GnRH pulses, but not the effects of 8Br-cAMP or forskolin. In conclusion, lowering intracellular cAMP levels with flufenamic acid attenuated GnRH-stimulated gonadotropin secretion, decreased alpha-subunit and FSH beta mRNA levels, and blocked the effect of GnRH to lengthen LH beta mRNA, whereas 8Br-cAMP or forskolin produced the opposite effect. These data extend previous results which suggested that cAMP modulates gonadotropin secretion and indicate that the cAMP/A-kinase pathway regulates each of the gonadotropin subunit mRNAs. PMID- 8404652 TI - Rapid down-regulation of c-jun protooncogene transcription by progesterone in the avian oviduct. AB - Previous work in this and other laboratories has shown that steroids rapidly regulate the expression of nuclear protooncogenes. In this present study, we have investigated the effect of progesterone (Pg) on the expression of c-jun in the avian oviduct system and its promoter activity in avian liver cells. Pg treatment of estrogen-withdrawn chickens brings about a decrease in the steady state mRNA level of the protooncogene c-jun within 30 min. This decrease is steroid dose dependent and gene specific. Using nuclear run-off transcription analyses, this rapid regulation was shown to occur at the level of gene transcription, as the rate of c-jun transcription decreases by more than 80% within 15 min after progesterone treatment. As expected, ovalbumin gene transcription is increased only after a lag period of 4 h following Pg treatment. In other studies, we have linked the c-jun promoter sequences between -1000 and +192 to chloramphenicol acetyltransferase reporter gene and cotransfected them into transformed avian liver cells along with the expression vector for the Pg receptor. Pg treatment of these cells causes a decrease in chloramphenicol acetyltransferase gene expression, albeit to a lesser extent than Pg inhibition of c-jun gene transcription. These results suggest that the 5'-domain of the chicken c-jun gene contains sequence elements that negatively regulate c-jun promoter activity in response to Pg. PMID- 8404653 TI - Regulation of prolactin, thyrotropin subunit, and gonadotropin subunit gene expression by pulsatile or continuous calcium signals. AB - We investigated the importance of calcium (Ca2+) influx in increasing the steady state concentrations of mRNAs coding for the pituitary peptides PRL, alpha, and TSH, LH, and FSH beta-subunits. Adult female rat pituitaries were dissociated, plated for 48 h, then inserted into perifusion chambers. Secretory responses were measured after 2 and 22 h of perifusion, and after 24 h, the cells were recovered, total RNA was extracted, and mRNAs were assayed by dot blot hybridization. The first experiment examined the effect of the Ca2+ channel blocker verapamil (100 microM) on the stimulatory action of pulsatile TRH (4 nM; 60-min interval) or GnRH (100 pM; 60-min interval) on pituitary mRNAs. TRH pulses induced a significant increase (49-56%) in PRL, alpha, and TSH beta mRNAs. Similarly, GnRH pulses stimulated a rise in alpha (64%) and FSH beta (50%) mRNAs, but not LH beta. The effects of pulsatile TRH or GnRH were eliminated when verapamil was added to the medium, suggesting that Ca2+ influx is critical to the stimulatory action of TRH or GnRH. The second experiment examined the effect of pulsatile vs. continuous increases in intracellular Ca2+ on pituitary mRNA expression. Pulsatile Ca2+ signals were produced by giving 60-min pulses of 50 mM KCl, Bay K 8644 (10 microM), or Bay K 8644 (10 microM; in the presence of 10 mM KCl in the injectate) and vehicle pulses to controls. Continuous increases in intracellular Ca2+ were induced by perifusion with medium containing the Ca2+ ionophore A23187 (20 microM), and these groups were compared to that receiving continuous verapamil. Pulsatile increases in Ca2+ influx (KCl or Bay K 8644) stimulated significant elevations in all mRNAs studied (36-74% increase vs. controls), with the exception of TSH beta. The magnitude of the mRNA responses to pulsatile Ca2+ (vs. controls) was similar to that observed after TRH and GnRH pulses. In contrast, only LH beta was increased by A23187 (42% increase vs. controls; P < 0.05). PRL and alpha mRNAs were selectively diminished by A23187 (57% and 83% decreases vs. controls, respectively; P < 0.05) and verapamil (67% and 60%; P < 0.05). The data show that expression of these pituitary genes is regulated by Ca2+ and that a pulsatile Ca2+ signal is required to stimulate PRL, alpha, and FSH beta (but not LH beta).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404654 TI - Recombinant human follicle-stimulating hormone is capable of exerting a biological effect in the adult hypophysectomized rat by reducing the numbers of degenerating germ cells. AB - There is considerable controversy as to whether FSH can, under normal circumstances, exert an effect to promote spermatogenesis in the adult rat. Recombinant human FSH (rhFSH) was used to answer a more limited question relating to whether FSH is capable of exerting a biological effect in promoting adult spermatogenesis. Can a pure preparation of FSH prevent the regressive changes seen after hypophysectomy (Hx) in a short term experiment? To answer this question, five groups of adult rats were used as follows: pituitary-intact animals, 3-day hypophysectomized (Hx), 3-day Hx given 3 mg testosterone propionate (T)/day for 7 days, 3-day Hx given 22 IU rhFSH for 7 days, and 3-day Hx given saline vehicle for 7 days. Testis weight, seminiferous tubule diameter, analysis of four degenerating germ cell types, the relative amount of lipid, and the levels of FSH receptors showed that FSH could, in a significant manner, prevent the regressive changes accompanying Hx. FSH was not as effective as T in doing so, because the FSH values were always intermediate between T-maintained animals and those after long term Hx. The Leydig cell was eliminated as a possible source of FSH-stimulated T promotion of spermatogenesis, given that morphometry and tissue T assays indicated that no additional production of T was elicited by rhFSH. The assay system used to enumerate degenerating germ cells proved a very sensitive indicator of the ability of hormones to maintain cell viability in short term experiments. The data not only show that FSH can exert a biological effect, but that this effect is qualitatively similar to that seen after the administration of T in terms of the maintenance of viability of specific germ cell types. A hypothesis is presented whereby FSH and T, although the former acting by a second messenger system and the latter by binding to nuclear receptors, can stimulate the genome to exert similar qualitative effects promoting the viability of germ cells. PMID- 8404655 TI - Failure of gonadotropin-releasing hormone (GnRH) pulses to increase luteinizing hormone beta messenger ribonucleic acid in GnRH-deficient female rats. AB - Gonadotropin subunit gene transcription and messenger RNA (mRNA) levels are differentially regulated by GnRH pulse frequency and amplitude in the male rat. The rapid changes of subunit mRNA levels and LH and FSH secretion during the estrous cycle, particularly the rapid rise in LH-beta subunit mRNA on proestrus afternoon, suggest that physiological changes in the pattern of GnRH action may also be important in female rats. However, in the absence of a GnRH-deficient female model the role of varying GnRH stimulation remains to be determined. We have characterized a GnRH-deficient model by administering the alpha-adrenergic antagonist phenoxybenzamine (PBZ) to ovariectomized (OVX) rats. Initial experiments showed that PBZ given 24 h earlier abolished the afternoon LH surge in OVX estradiol (E2) replaced rats whereas LH responses to exogenous GnRH were preserved. A PBZ regimen of 15 mg/kg ip at OVX followed by 10 mg/kg at 24 h and 5 mg/kg at 48 h prevented the increase in alpha, LH-beta, and FSH-beta mRNAs and LH and FSH secretion for 72 h post-OVX. LH and FSH responses to GnRH pulses were preserved suggesting that PBZ blocked the post-OVX increase in hypothalamic GnRH secretion. The suppressive effect of PBZ appeared to be specific to the hypothalamic-pituitary-ovarian axis as plasma PRL, TSH, and corticosterone were not decreased compared to controls. We have used this GnRH-deficient OVX female model to investigate the effects of exogenous GnRH pulses on subunit mRNA expression. GnRH pulses (5-250 ng/30 min for 12-24 h) were administered via an intraatrial catheter beginning 24 h after OVX and the first PBZ injection (OVX+PBZ+saline pulses to controls). Expression of alpha and FSH-beta mRNAs and LH and FSH secretion were increased by GnRH pulse doses of 5-25 ng to values similar to or greater than those in OVX controls though the higher doses of GnRH/pulse did not increase FSH-beta mRNA or plasma FSH. However, LH-beta mRNA levels were not increased by GnRH pulses. GnRH pulses were also given to rats replaced with proestrus concentrations of estradiol alone or in combination with progesterone (P). Again, no demonstrable increases in LH-beta mRNA expression were observed. alpha-mRNA concentrations were further increased in the presence of E2 alone, and P in combination with E2, produced an augmented response of FSH beta subunit mRNA. These data suggest that ovarian steroid hormones act directly on the gonadotrope to augment alpha and FSH-beta mRNA responses to GnRH.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404656 TI - Changes in pituitary gonadotropin-releasing hormone receptor messenger ribonucleic acid content during lactation and after pup removal. AB - Lactation is associated with a reduction in pituitary GnRH receptor content (GnRH R) when compared with diestrus of the estrous cycle. These studies examined whether the changes in GnRH-R during lactation involved alterations in GnRH-R gene expression. GnRH-R messenger RNA (mRNA) levels were assessed by in situ hybridization, using a 35S-labeled antisense riboprobe coding for the transmembrane region of the mouse GnRH-R gene. The area occupied by grains in 20 microns sections of the pituitary was assessed with an image analysis program. The specificity of the probe for GnRH-R mRNA was demonstrated by the presence of only background grains over the pituitary when a sense riboprobe to GnRH-R mRNA was used. Also, there was an absence of clusters of grains over the posterior pituitary, an area devoid of gonadotropes. In lactating rats suckling eight pups on day 10 postpartum, GnRH-R mRNA levels in the anterior pituitary were suppressed by 60% when compared with the levels observed during diestrus. Removal of the eight-pup suckling stimulus for 24 h reestablished the diestrous pattern of GnRH-R mRNA expression. These results demonstrate that the suckling stimulus results in a significant suppression of GnRH-R mRNA levels that is reversed by removal of the stimulus. These changes in GnRH-R mRNA most likely contribute to the changes in GnRH-R content observed in response to the suckling stimulus. PMID- 8404657 TI - Protein phosphatase-1 and -2a activities in cultured fetal chick neurons: differential regulation by insulin and insulin-like growth factor-I. AB - In this study, we examined the developmental expression and regulation by insulin and insulin-like growth factor-I (IGF-I) of protein phosphatase-1 (PP-1) and protein phosphatase-2A (PP-2A) in cultured fetal chick neurons. Protein phosphatase activities were measured using 32P-labeled phosphorylase-a or 32P labeled S6 kinase substrate peptide. In cell extracts from day 1-5 cultures, 40 45% of spontaneous protein phosphatase activity was due to PP-1. PP-2A accounted for the remaining 55-60% of enzyme activity. Spontaneous PP-1 activity increased by 100% in day 2 cultures and remained constant thereafter. PP-2A activity increased by 48% in day 2 cultures, with minimal increases in enzyme activity in later cultures. Under the assay conditions employed, at all times in culture a significant proportion (45-50%) of PP-1 was in an inactive form that could be reactivated by trypsin. PP-2A activity was not influenced by trypsin. Insulin stimulated neuronal PP-1 activity in day 4 and 5 cultures, but had no effect in earlier cultures. The activation of PP-1 by insulin was rapid, with a maximal effect (30-40% increase over basal levels) at 5 min with 10 ng/ml insulin. Insulin did not alter total (trypsin-released) PP-1 activity, the content of PP-1 catalytic subunit, or PP-2A activity at any time in culture. In contrast to insulin, IGF-I had no effect on PP-1 activity at any time in culture, but significantly increased PP-2A activity in day 5 cultures. Maximal stimulation of PP-2A activity by IGF-I was observed at 10 min, with an EC50 of 5 ng/ml. These results indicate that chick forebrain neurons contain both PP-1 and PP-2A activities and that neuronal PP-1 and PP-2A activities are differentially regulated by insulin and IGF-I. We conclude that although insulin and IGF-I share many steps in signal transduction, these growth factors have distinct actions on neuronal phosphatase activity that may impact on differences in their neurotropic actions. PMID- 8404658 TI - Hyperprolactinemia suppresses the luteinizing hormone responses to N-methyl-D aspartate, epinephrine, and neuropeptide-Y in male rats. AB - This study characterizes the responses of LHRH neurons to N-methyl-D-aspartate (NMDA), norepinephrine, epinephrine (E), and neuropeptide-Y (NPY), as evidenced indirectly by the measurement of circulating LH titers, and investigates whether neurons using these compounds as neurotransmitters might be involved in mediating hyperprolactinemic (HP) suppression of LH release. Male rats were orchidectomized, adrenalectomized, and implanted with a testosterone-containing Silastic capsule, a 50% corticosterone pellet, and third cerebroventricular and right atrial cannulae at time zero. Rats received sc injections of ovine PRL (2400 micrograms/250 microliters) in a polyvinylpyrrolidone depot or vehicle every 12 h for 48 h when experiments were performed. The mean maximal LH increments (delta LH) in response to two doses of LHRH (0.4 and 0.8 ng/100 g BW) were not altered in HP rats, indicating that ovine PRL did not cause a change in pituitary responsiveness. NMDA (20 mg/kg BW, iv)-induced LH release peaked 5 min after injection. The delta LH (0-5 min) in HP rats was suppressed by 53% compared with the control value. Epinephrine [5, 10, and 15 micrograms/2 microliters, intracerebroventricularly (icv)], but not norepinephrine (20 and 40 micrograms/2 microliters, icv), produced dose-dependent LH responses that peaked at 10 min. The delta LH (0-10 min) in HP rats in response to 10 micrograms/2 microliters E was suppressed by 68% compared with the control value. Two doses of NPY (2 and 10 micrograms/2 microliters, icv) produced dose-dependent LH increments that peaked at 10 min. In HP rats, the delta LH (0-10 min) in response to 10 micrograms/2 microliters NPY was suppressed 52% compared with the control value. The combined administration of E (10 or 16 micrograms) and NPY (5 or 10 micrograms) produced mean maximal LH responses that significantly exceeded the additive responses of these compounds individually. This synergistic effect may be mediated by separate adrenergic and NPYergic afferents to the LHRH neurons or may, in fact, reflect corelease of these two neurotransmitters from the same neurons. The LH responses to NMDA, E, and NPY were all inhibited in HP rats. This suggests that elevated PRL levels act on the LHRH neurons, either directly or indirectly through an inhibitory afferent neuronal system, to decrease their responsivity to all stimuli. PMID- 8404659 TI - A polyclonal antibody to a synthetic peptide derived from the rat follicle stimulating hormone receptor reveals the recombinant receptor as a 74-kilodalton protein. AB - We have prepared a polyclonal antibody (AntiF) against a synthetic peptide comprising residues 19-29 of the rat FSH receptor. The specificity of this antibody was documented using human embryonic kidney (293) cells and stable transfectants of 293 cells expressing the recombinant LH/CG and FSH receptors. The data presented show that AntiF inhibits the binding of FSH, but not that of LH/CG, to their cognate receptors. AntiF also recognizes a specific protein(s) representing the FSH receptor in Western blots or by immunoprecipitation of cells transfected with the FSH receptor. This protein(s) is not present in untransfected 293 cells or in 293 cells permanently transfected with the LH/CG receptor. Finally, addition of the 19-29 peptide prevents immunoprecipitation of the FSH receptor by AntiF, whereas an unrelated peptide corresponding to residues 637-647 has no effect. In Western blots of 293 cells transfected with the FSH receptor, AntiF reveals the recombinant receptor as a heterogenous glycoprotein with a molecular mass of 58,000-83,000 daltons (in the absence of thiol-reducing agents) or 69,000-81,000 (in the presence of thiol reducing agents). Pulse-chase experiments with metabolically labeled cells show that the mature FSH receptor is a 74-kilodalton protein derived from 67- and 72-kilodalton precursors. PMID- 8404660 TI - Role of protein kinase-C in the alpha 1-adrenoceptor-mediated responses of perfused rat liver. AB - The present work aimed to determine the role played by protein kinase-C (PKC) in the alpha 1-adrenoceptor-induced activation of hepatic metabolism. The following observations indicate that activation of PKC is a condition necessary for alpha 1 adrenoceptor activation of hepatic functions, but not sufficient to mimic the receptor-mediated effects in the absence of external physiological stimuli. 1) alpha 1-Adrenoceptor activation promoted the translocation of PKC from the cytosol to its active form in the plasma membrane. 2) Activation of PKC by the phorbol ester 12-myristate 13-acetate or exogenous diacylglycerols or by elevation of endogenous levels of diacylglycerols by inhibiting diacylglycerol kinase mimicked the alpha 1-adrenoceptor-mediated actions. However, the time course and magnitude of the nonreceptor responses differ from those mediated by alpha 1-adrenoceptor activation. In addition, nonreceptor-mediated activation of PKC decreased the alpha 1-adrenoceptor responsiveness. 3) Inhibition of PKC by either H-7 [1-(5-isoquinolinilsulfonyl)2-methylpiperazine] or staurosporine inhibited all of the alpha 1-adrenoceptor-induced responses, except gluconeogenesis. The vasopressin effects were not inhibited by H-7, indicating that PKC activation is a distinct feature of the hepatic alpha 1-adrenoceptor activation that is not shared by all the Ca(2+)-mobilizing agonists. The diacylglycerol-PKC branch of the alpha 1-adrenoceptor signaling pathway seems to control the sustained phase of stimulation of hepatic functions. In these studies we have also observed that phorbol 12-myristate 13-acetate produces a concentration-dependent inhibition of hepatic respiration. However, decreased energy availability does not seem to be the cause of its action to decrease alpha 1-adrenoceptor responsiveness. PMID- 8404661 TI - Thyroxine transport to the brain: role of protein synthesis by the choroid plexus. AB - A cell culture model for the blood-cerebrospinal fluid barrier in choroid plexus was developed. The relationship between synthesis and secretion of transthyretin across a layer of epithelial cells derived from rat choroid plexus and the transport of T4 was analyzed in a two-chamber system. Choroid plexus cells were dispersed and placed on a porous filter suspended in cell culture medium. A monolayer of polarized epithelial cells developed after 5 days in culture, separating fluid in the upper (apical) chamber from fluid in the lower (basal) chamber. Electrical resistance across the cell layer was 100 Ohm/cm2. Transthyretin was synthesized and secreted by these cells. Over 32 h, transthyretin accumulated in the fluid in the apical chamber to twice the concentration in the basal chamber. [125I]T4 added to the basal chamber permeated to the apical fluid and accumulated in the apical chamber to twice the concentration in the basal fluid. Upon inhibition of protein synthesis, T4 equilibrated to a similar concentration in the two chambers. Thus, the accumulation of T4 in the apical chamber required continuing protein synthesis. Competitive inhibition of T4 binding to transthyretin by EMD 21388 also prevented the accumulation of T4 to a higher concentration in the upper than in the lower chamber. These data suggest that T4 partitions through the choroid plexus and that transthyretin synthesis and secretion by the choroid plexus determines the concentration of T4 in the apical fluid. A model is proposed for the involvement of transthyretin secreted by the choroid plexus, in the in vivo distribution of T4 in the brain. PMID- 8404662 TI - Possible involvement of protein kinase C in gonadotropin-induced ovulation in the rat ovary. AB - Recent reports indicate that protein kinase C may play an important role in the process of gonadotropin-induced ovulation in the ovary. In the present study, we examined the effect of the protein kinase C inhibitor, 1-(5-isoquinolinesulfonyl) 2-methylpiperazine (H-7), on LH-stimulated tissue type plasminogen activator (tPA) activity in cultured rat granulosa cells. Granulosa cells were obtained from PMSG-treated rats and cultured for 48 h in the presence or absence of H-7 (0.1-60 microM) with ovine LH (30 ng/ml), phorbol 12-myristate 13-acetate (10(-8) M), phorbol 12,13-dibutyrate (10(-8) M), or (Bu)2cAMP (5 mM). After culture, tPA activity in the conditioned medium was assayed by fibrin autography technique after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. H-7 (1.0-60 microM) inhibited LH-, phorbol 12-myristate 13-acetate-, or phorbol 12,13 dibutyrate-stimulated tPA activity dose dependently, and each ID50 was approximately 8 microM. However, H-7 did not inhibit (Bu)2cAMP-stimulated tPA activity. To investigate the effect of H-7 on the ovulatory process in vivo, PMSG treated immature rats were injected with H-7 (10(-9)-10(-3) M) into the unilateral ovarian bursa just before human CG administration. After 24 h, the number of oocyte-cumulus complexes in the oviduct was counted. H-7 suppressed the number of oocytes released from treated ovaries dose-dependently. The light microscopical observation revealed that ovaries treated with H-7 contained a few corpora lutea and many large unruptured follicles. The results of the present study suggest that the suppressive effects of H-7 on human CG-induced ovulation might be partly due to the inhibition of tPA secretion by rat granulosa cells via protein kinase C inhibition. PMID- 8404663 TI - Differential expression of type I adrenal steroid receptors in immune tissues is associated with tissue-specific regulation of type II receptors by aldosterone. AB - We examined the influence of the type I adrenal steroid receptor agonist, aldosterone, on type II adrenal steroid receptor binding in the rat spleen and thymus after adrenalectomy. In the spleen, adrenalectomy was associated with a significant up-regulation of type II receptors, which was blocked by the concurrent administration of aldosterone (1 microgram/h) via sc osmotic minipumps. Neither adrenalectomy nor aldosterone treatment altered type II receptor binding in the thymus. Despite high doses of aldosterone (10 micrograms/h), which resulted in supra-physiological blood concentrations of this hormone, there was no evidence of type II receptor decreases in spleen or thymus below receptor levels found in sham-adrenalectomized rats. The effect of aldosterone on type II receptor binding appeared to be mediated by the type I receptor, since there was no aldosterone effect on the thymus, which did not exhibit detectable levels of type I receptor binding. Moreover, there was no evidence that aldosterone competed for the type II receptor in vivo or in vitro as determined by measurements of the type II receptor dissociation constant in the spleen of adrenalectomized, aldosterone-treated animals. Since selective activation of the type I receptor occurs in the spleen under physiological conditions, these results indicate that type I receptors may play a tonic inhibitory role in type II receptor expression in immune cells which express both receptor subtypes and reside in this tissue. Furthermore, the findings suggest that there may be different mechanisms involved in the up-regulation vs. the down regulation of type II adrenal steroid receptors, and effects mediated solely via the type I adrenal steroid receptor appear only to influence the former process. PMID- 8404664 TI - Receptor-mediated stimulatory effect of atrial natriuretic factor, brain natriuretic peptide, and C-type natriuretic peptide on testosterone production in purified mouse Leydig cells: activation of cholesterol side-chain cleavage enzyme. AB - We have investigated the mechanism by which different natriuretic peptides stimulate steroidogenesis in purified mouse Leydig cells. In addition to atrial natriuretic factor (ANF), we show that brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP) also stimulate testosterone production in these cells. Testosterone production was increased dramatically to 14-fold with ANF (EC50 = 0.3 nM) and 15-fold with BNP (EC50 = 0.2 nM); however, the CNP-stimulated level of testosterone production was only 2.5-fold compared with controls. ANF and BNP enhanced the stimulatory effect of LH on testosterone production. The C-ANF(4-23) (a truncated form of ANF) had no effect on testosterone production in these cells. ANF, BNP, and CNP stimulated the production of intermediate precursors of testosterone biosynthesis, which included progesterone, 17 alpha-hydroxy progesterone, androstenedione, pregnenolone, 17 alpha-hydroxy pregnenolone, and dehydroepiandrosterone sulfate. The conversion of pregnenolone and progesterone to testosterone was also significantly enhanced after treatment of Leydig cells with these peptides. All three natriuretic peptides (ANF, BNP, and CNP) stimulated the activity of particulate guanylate cyclase by 8.4-, 8.5-, and 4.8 fold and the accumulation of intracellular cGMP by 52-, 58-, and 19-fold, respectively. The cGMP inhibitor LY83583 attenuated both the generation of cGMP as well as testosterone in response to these natriuretic peptides, suggesting the involvement of cGMP as a second messenger. Leydig cells were found to contain high affinity and low capacity binding sites for ANF [dissociation constant (Kd), 2.0 x 10(-10) M; maximum binding capacity (Bmax). 20 fmol/1 x 10(5) cells], BNP (Kd, 2.2 x 10(-10) M; Bmax, 19 fmol/1 x 10(5) cells), and CNP (Kd, 3.1 x 10(-10) M; Bmax, 8.6 fmol/1 x 10(5) cells). The results presented here document that a family of different natriuretic peptides stimulates Leydig cell steroidogenesis in receptor-mediated fashion, beginning at the cholesterol side-chain cleavage enzyme. The data also show that these peptide hormones induce testosterone production in mouse Leydig cells by involving both delta 4- and delta 5-pathways of steroidogenesis. PMID- 8404665 TI - Pituitary folliculo-stellate-like cell line (TtT/GF) responds to novel hypophysiotropic peptide (pituitary adenylate cyclase-activating peptide), showing increased adenosine 3',5'-monophosphate and interleukin-6 secretion and cell proliferation. AB - Several studies have shown that folliculo-stellate cells (FS cells) in the anterior pituitary gland exhibit paracrine functions. Recently, we established a pituitary FS-like cell line, TtT/GF, which was derived from an isologously transplantable pituitary thyrotropic tumor line induced by radiothyroidectomy. In studies to examine the function of FS cells, we found that two forms of a novel hypophysiotropic peptide, pituitary adenylate cyclase-activating polypeptide (PACAP), were potent activators of TtT/GF cells. Both the 27- and 38-amino acid forms of PACAP (PACAP-27 and PACAP-38) and vasoactive intestinal peptide (VIP) increased the levels of cAMP in TtT/GF cells in a similar dose-dependent manner. PACAP-27 and PACAP-38 specifically stimulated the proliferation of TtT/GF cells dose dependently, whereas VIP was ineffective. The minimal effective concentration of the PACAPs inducing cell proliferation was between 10(-8)-10(-7) M. However, PACAP-27 was much less potent than PACAP-38 in stimulating cell proliferation and DNA synthesis. PACAP-38, PACAP-27, and VIP all stimulated the release of interleukin-6 (IL-6) from TtT/GF cells. PACAP 38 (10(-8) M) stimulated IL-6 production effectively within 1 h of incubation, and the level attained at 8 h of cultivation (620 pg/ml) was nearly 10-fold that in the absence of PACAP-38 (60 pg/ml). PACAP-38 and VIP stimulated IL-6 secretion significantly at 10(-10) 10(-9) M in a bell-shaped manner; the maximum values were 10(-7) and 10(-8) M, respectively. On the other hand, IL-6 secretion stimulated by PACAP-27 became saturated at 10(-8) M, and the maximum value (320 pg/ml) was about 25% of that stimulated by PACAP-38 (1280 pg/ml). These findings obtained using TtT/GF cells as a model of FS cells suggest that PACAP acts as a hypophysiotropic factor, which targets FS cells and stimulates their proliferation, adenylate cyclase activation, and IL-6 secretion. PMID- 8404666 TI - Dephosphorylation of standard prolactin produces a more biologically active molecule: evidence for antagonism between nonphosphorylated and phosphorylated prolactin in the stimulation of Nb2 cell proliferation. AB - Charge isomers of monomer PRL have been described for all species thus far examined. For the rat, cow, and chicken, at least a major proportion of the more acidic isomers have been shown to be phosphorylated. The physiological relevance of this posttranslational phosphorylation, however, remains unclear. In this study we have compared the growth-promoting activities of dephosphorylated and standard rat PRL (partially phosphorylated) in the widely used Nb2 bioassay. Dephosphorylated PRL was produced by treatment of standard rat PRL with acid phosphatase, resulting in the conversion of a 91.9% nonphosphorylated/8.1% phosphorylated preparation to a 98.6% nonphosphorylated preparation. These two preparations were added separately or in combination to stationary Nb2 cells. Cell number was assessed 3 days later using a colorimetric assay. Dephosphorylated PRL showed significantly higher growth-promoting activity than standard PRL at concentrations up to 10 ng/ml. In the 1-5 ng/ml concentration range, dephosphorylated PRL was twice as active. Given that the conversion of 6.7% phosphorylated PRL to nonphosphorylated PRL resulted in a doubling of activity, one can deduce not only that phospho-PRL acted as an antagonist to nonphosphorylated PRL in this assay, but also that it did not do so on an equimolar basis. Titration of the two PRL preparations produced data confirming this latter deduction. These data are of importance in our understanding of PRL promoted cell proliferation. PMID- 8404667 TI - Temporal and stage-specific changes in spermatogenesis of rat after gonadotropin deprivation by a potent gonadotropin-releasing hormone antagonist treatment. AB - GnRH antagonists (GnRH-As) rapidly and reversibly inhibit testicular functions in a variety of experimental models as well as man. Their potential for human male contraception is currently being tested in many centers, including our own. This study was undertaken to provide comprehensive quantitative information on the testes and to document the temporal and stage-specific changes in the kinetics of germ cell degeneration in rats treated daily with the Nal-Glu GnRH-A (1250 micrograms/kg body wt) for up to 4 weeks. Plasma levels of testosterone (T) and the concentrations of testicular T declined to 20.7% and 5.4% of control values, respectively, by 1 week and remained suppressed throughout the treatment period. Preleptotene and pachytene spermatocytes, and step-7 and step-19 spermatids at stage VII were the first germ cells to degenerate soon after (1 week) GnRH-A treatment. Germ cell counts at stage VII also revealed a significant (P < 0.05) reduction in number of preleptotene (25.6%), pachytene (35.4%), and step-7 spermatids (29.1%) in comparison with controls. The number of homogenization resistant advanced spermatids decreased by 70%. A further progressive loss of spermatogenic activity occurred with time. Treatment with GnRH-A for 4 weeks caused advanced spermatids to decline to nearly undetectable, Step-7 spermatids to decline to 17.7% of the normal level, and the P and PL to decline to 28.6% and 67.7%, respectively, of control values. The number of Sertoli cells and A1 spermatogonia remained unchanged throughout the experimental period. The effects of GnRH-A treatment on spermatogenesis were identical to that of hypophysectomy. These results suggest that: 1) early deprivation of gonadotropins and/or intratesticular T by GnRH-A treatment is followed by stage-specific degeneration of germ cells; 2) pituitary secretions other than LH and FSH have little primary influence on spermatogenesis during early regression; and 3) the GnRH-A-treated rat would be an excellent animal model for studying the targeted effects of LH, FSH, and T on the regulation of spermatogenesis. PMID- 8404668 TI - 9-cis-retinoic acid regulates the expression of the muscle determination gene Myf5. AB - Myf5 is a member of the MyoD family, a set of four helix-loop-helix transcription factors that controls myogenic differentiation. The Myf5 gene has both in vivo and in vitro expression patterns consistent with an involvement in the first events of myogenesis, such as acquisition and/or maintenance of myogenic "determined" phenotype. To date, very little is known about the mechanism underlying the tight regulation of Myf5 expression. We report here that retinoic acid (RA) reduces the level of Myf5 message in both mouse C2 and rat L6 cell lines, probably at the transcriptional level, because Myf5 mRNA stability is not affected by RA. This repression is dose dependent, starting at 0.1 microM of all trans RA, and is not abrogated by cycloheximide, suggesting a direct involvement of RA receptors in the control of Myf5 expression. Furthermore, we compared the efficiency of natural (all-trans RA and 9-cis RA) or synthetic (TTNPB) retinoids that differentially activate the two families of RA receptors, RA receptors and retinoid X-receptors (9-cis RA). As 9-cis RA is about 10 times more efficient than all-trans RA in repressing Myf5, whereas TTNPB, which preferentially activates RA receptors, is far less potent, our data provide evidence for an important role of ligand-bound retinoid X-receptors in the mediation of this inhibition. PMID- 8404669 TI - Glucuronidation of thyroid hormone in rat liver: effects of in vivo treatment with microsomal enzyme inducers and in vitro assay conditions. AB - We investigated the effects of in vivo treatment with different microsomal enzyme inducers, including clofibrate (CLOF), hexachlorobenzene (HCB), 3 methylcholanthrene (MC), 3,3',4,4'-tetrachlorobiphenyl (TCB), and 2,3,7,8 tetrachloro-p-dioxin, as well as of in vitro addition of the detergent Brij 56 on the glucuronidation of T4, T3, and rT3 by UDP-glucuronyltransferase (UGT) activities of rat liver microsomes. The results were compared with measurements of UGT activities for bilirubin, p-nitrophenol (PNP), and androsterone. In general, glucuronidation rates were 5-fold or more higher with rT3 than with T4 or T3 as substrate. In liver microsomes from untreated rats, T4 UGT activity was stimulated by Brij 56 to a maximum of about 2-fold at 0.025% detergent. Treatment of Wistar rats for 4 days with CLOF (200 mg/kg BW.day) resulted in significant increases in UGT activities for T4 (to 154%), rT3 (to 155%), and bilirubin (to 194%), in particular if assayed in the presence of 0.025% Brij 56, but had little effect on the UGT activities for T3, PNP, and androsterone. The CLOF-induced increases in T4 and rT3 UGT activities were not observed in Gunn rats, which have a complete lack of bilirubin UGT activity and greatly impaired PNP UGT activity. Treatment of Wistar rats with a single injection of MC (50 mg/kg BW), TCB (50 mg/kg BW), or 2,3,7,8-tetrachloro-p-dioxin (6.25 micrograms/kg BW) resulted, after 4 days, in 6.3- to 7.3-fold increases in T4 UGT activity and 15.1- to 16.7 fold increases in rT3 UGT activity if determined in the absence of Brij 56, whereas T4 UGT activity was only increased by 33-68% when assayed in the presence of Brij 56. T3 glucuronidation was not affected (with Brij 56) or was increased by only 33-68% (without Brij 56) after treatment with these MC-type inducers. PNP UGT activity was induced 3.6- to 4.3-fold, whereas bilirubin and androsterone UGT activities were changed little by these treatments. Similar findings regarding T4, rT3, PNP, and bilirubin UGT activities were obtained after chronic treatment of WAG rats with HCB, another MC-type inducer. However, WAG rats lack androsterone UGT and show low T3 UGT activity, which was increased about 2.3-fold by HCB treatment. On the basis of these and previous findings it is concluded that at least three UGT isoenzymes are involved in the glucuronidation of thyroid hormone.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8404670 TI - Glucocorticoid-induced activation of latent transforming growth factor-beta by normal human osteoblast-like cells. AB - Transforming growth factor-beta (TGF beta) is a multifunctional growth factor in bone that is secreted as a latent complex and must be activated in order to influence cellular activity. We have investigated the influence of dexamethasone (Dex; a potent glucocorticoid) on the secretion and activation of latent TGF beta by normal human osteoblast-like cells (hOBs). Dex had no significant effect on TGF beta mRNA or total protein production, but treatment with Dex resulted in a steroid dose-dependent activation of up to 90% of the TGF beta produced by the hOBs. Dex-treated hOBs activated multiple latent forms of TGF beta. Conditioned medium from Dex-treated hOBs retained the ability to activate latent TGF beta when incubated at 37 C in the absence of cells. Unlike other cell systems, Dex induced hOB-mediated TGF beta activation did not involve binding to the mannose-6 phosphate receptor. However, the activation was prevented by treatment of hOB cells with microtubual disrupting agents, by the addition of protease inhibitors, or by weak base treatment of the medium. Dex treatment of hOBs was shown to induce a dose-dependent increase in the mRNA levels of cathepsin-B and -D and in the levels of cathepsin-B protein secreted by the cells. Taken together, these data suggest that Dex treatment of hOBs induces the production and secretion of lysosomal proteases that, when secreted, activate latent TGF beta which is secreted by the hOB cells. There is evidence for an involvement of more than one type of protease in this activation process. This activation of TGF beta may, therefore, play a role in glucocorticoid regulation of bone cell functions. Furthermore, TGF beta is most likely involved in autocrine and paracrine effects on bone cells. PMID- 8404671 TI - Cyclic adenosine 3',5'-monophosphate induces prolactin expression in stromal cells isolated from human proliferative endometrium. AB - In vitro decidualization of stromal cells isolated from human proliferative endometrium and cultured in RPMI-1640 containing 2% charcoal-treated fetal bovine serum and 0.1 U/ml insulin was achieved by adding to the medium cAMP derivatives [(Bu)2cAMP (db-cAMP) and 8-bromo-cAMP] or forskolin. PRL production under these conditions was demonstrated by documenting the synthesis of PRL mRNA (approximately 1.1 kilobase), the output of immunoprecipitable [35S]methionine labeled PRL migrating as a single 23-kilodalton band during gel electrophoresis, and the time- and concentration-dependent secretion of PRL into the medium, measured by RIA (maximal on days 4-5 using 0.5 mM db-cAMP). Medroxyprogesterone acetate (1 microM) enhanced (1.7- to 2.5-fold) the effect of db-cAMP, 8-bromo cAMP, or forskolin on PRL production, as evaluated by Western blotting analysis. Further evidence for a participation of db-cAMP in the decidualization process was provided by its ability to induce immunocytochemically detectable heat shock protein-27, insulin-like growth factor-binding protein-1, desmin, and laminin, all compounds produced by human decidual cells, but not expressed by stromal cells. The induction of PRL by cAMP may be a key step in the process of differentiation of fibroblast-like stromal cells to the decidual phenotype, as it has been previously reported by this laboratory that, under similar culture conditions, PRL itself is capable of inducing the production of heat shock protein-27, insulin-like growth factor-binding protein-1, desmin, and laminin in stromal cells isolated from proliferative endometrium. PMID- 8404672 TI - Estrogens inhibit and androgens enhance ovarian granulosa cell apoptosis. AB - Apoptotic cell death has recently been suggested to be the underlying mechanism of ovarian follicle atresia. To study the regulation of follicle cell apoptosis by sex steroids, we have analyzed ovarian DNA fragmentation, the hallmark of apoptosis, in rats treated with estrogens and androgens. Immature rats were hypophysectomized and implanted with diethylstilbestrol (DES) capsules. Two days later, DES implants were removed in some animals, followed by treatment with estrogens with or without androgens. The extent of ovarian apoptotic DNA fragmentation was analyzed by autoradiography of size-fractionated DNA labeled at 3'-ends by [32P]dideoxy-ATP. After DES withdrawal, ovarian weight decreased and DNA fragmentation increased in a time-dependent manner. In granulosa cells, an increase in apoptotic DNA fragmentation was seen 12 h after withdrawal of DES implants, followed by a 25-fold increase at 48 h. In situ analysis of DNA fragmentation on histological sections of ovaries, using a nonisotopic labeling of DNA by digoxigenin-dideoxy-UTP, also demonstrated that apoptosis induced by DES withdrawal is confined to the granulosa cells in early antral and preantral follicles. No increase in DNA breakdown was detected in thecal cells and interstitial tissues or granulosa cells of primordial and primary follicles. In contrast, replacement with DES (0.5 mg twice daily) or estradiol benzoate (3 mg daily) completely prevented the observed ovarian weight loss and increases in granulosa cell apoptosis. Treatment with estradiol benzoate (0.003-3 mg/day) dose dependently suppressed the apoptosis seen 2 days after removal of DES implants. Furthermore, the antiatretogenic effect of estrogen was blocked by treatment with testosterone (0.5 mg twice daily), which increased ovarian apoptotic DNA fragmentation and decreased ovarian weight in DES-treated animals in a time dependent manner. Also, in situ examination showed that androgen treatment increased apoptosis in the granulosa cells in a subpopulation of early antral and preantral follicles. The specificity of testosterone action was further demonstrated by the lack of effect of progesterone and cortisol on ovarian apoptosis. These data suggest that sex steroids play an important role in the regulation of ovarian apoptotic cell death, with estrogens preventing apoptosis and androgens antagonizing the effect of estrogens. These data provide the basis for future studies on the role of sex steroid hormones in follicular atresia and the regulation of endonuclease activity by steroid hormones. PMID- 8404673 TI - Cytosolic free Ca2+ in human syncytiotrophoblast cells increased by gonadotropin releasing hormone. AB - Effects of GnRH on free cytosolic Ca2+ concentrations ([Ca2+]i) were examined in individual first trimester human cytotrophoblast and syncytiotrophoblast cells by fura-2 microspectrofluorimetry. GnRH (10(-6) M) did not affect [Ca2+]i in cytotrophoblasts on days 2-9 of culture, with 50 cells tested each day. GnRH (10( 6) M) did not affect [Ca2+]i on days 2-3, but increased [Ca2+]i in 15% of culture derived syncytiotrophoblasts on day 4 (8 of 52 cells) and in 48% on days 5-9 of culture (158 of 332 cells). Culture-derived syncytiotrophoblasts originated from four first trimester placentae. GnRH increased [Ca2+]i in a preliminary trial using syncytiotrophoblasts derived directly from a single first trimester placenta and cultured for 3 days (13 of 33 cells). Culture-derived first trimester syncytiotrophoblast cells that responded to 10(-6) M GnRH (28 of 63 cells) on day 6 also responded to GnRH at 10(-7) M (26 of 28 of the above cells), 10(-8) M (24 of 28 cells, 10(-9) M (22 of 28 cells), and 10(-10) M (3 of 28 cells). No cells responded to GnRH below a concentration of 10(-10) M. Desensitization of syncytiotrophoblasts by continuous GnRH perifusion (10(-6) M) and blockade of GnRH by competitive antagonism with Nal-Glu-GnRH (10(-6) M) suggested that effects of GnRH were receptor specific. The results provide direct evidence supporting the contention that the intracellular signaling resultant from GnRH receptor-ligand interactions in syncytiotrophoblasts may be at least partially mediated by transient increases in [Ca2+]i. PMID- 8404674 TI - Effects of steroid and lactogenic hormones on islets of Langerhans: a new hypothesis for the role of pregnancy steroids in the adaptation of islets to pregnancy. AB - Adaptive changes that occur in islets of Langerhans during pregnancy include enhanced insulin secretion, insulin synthesis, beta-cell proliferation, gap junctional coupling among beta-cells, and glucose oxidation. We have determined that elevated lactogenic activity is directly responsible for these changes in beta-cell function. Recently, we showed that two of the principal adaptive characteristics (insulin secretion and beta-cell proliferation) of rat pregnancy peaked on day 15 and returned to control levels by day 20. As placental lactogen remains elevated during late gestation, it was of interest to determine whether pregnancy steroids could reverse the effects of lactogen on islets. In this study, rat islets were cultured with progesterone, estradiol, rat PRL (rPRL), or combinations of these hormones (progesterone and rPRL, estradiol and rPRL, or progesterone and estradiol and rPRL). Insulin secretion was examined for 8 days, and beta-cell proliferation by 2-bromo-5'-deoxyuridine (BrdU) incorporation on days 4 and 8. rPRL treatment resulted in a time-dependent increase in insulin secretion that was 3-fold greater than that from control islets by day 8. Progesterone and estradiol had minimal effects on insulin secretion. Estradiol had no effect on the increased insulin secretion observed with rPRL during the first 6 days and a small inhibitory effect on days 7 and 8. Although progesterone treatment had no effect on the increased insulin secretion induced by rPRL during the first 3 days, it subsequently resulted in a decline in insulin secretion to that from control islets. The combination of progesterone and estradiol was more effective than either steroid by itself in reversing the effects of rPRL on insulin secretion. Similar results were obtained in the BrdU labeling experiments: 1) a 7-fold increase in the number of BrdU-labeled nuclei per islet was observed after culture in the presence of rPRL; and 2) estradiol had a small inhibitory effect on the increased BrdU labeling observed with rPRL; however, 3) progesterone completely reversed the effect of rPRL on islet beta-cell division. These results demonstrate that progesterone counterregulates the effects of PRL on insulin secretion and islet beta-cell division. The temporal changes observed in islets in vitro under the influence of PRL and progesterone mimic those seen in islets during pregnancy. We conclude that progesterone, which increases in the later stages of gestation, is the primary hormone responsible for counteracting the stimulatory effects of elevated lactogenic activity on islets during late pregnancy. PMID- 8404675 TI - Sodium restriction increases aldosterone biosynthesis by increasing late pathway, but not early pathway, messenger ribonucleic acid levels and enzyme activity in normotensive rats. AB - To determine whether changes in dietary sodium intake modify the early and/or late pathways of aldosterone biosynthesis, we studied in Sprague-Dawley rats the effect of sodium restriction on early (conversion of cholesterol to pregnenolone) and late (conversion of corticosterone to aldosterone) pathway activity and on the mRNA levels for the enzymes regulating these steps. Sodium restriction increased basal and angiotensin-II-stimulated aldosterone output from isolated zona glomerulosa cells by 5- to 9-fold. This increase in aldosterone output did not appear to be due to changes in the conversion of cholesterol to pregnenolone or in the mRNA levels of the early pathway enzyme, cholesterol side-chain cleavage cytochrome P-450. In contrast, sodium restriction increased the conversion of corticosterone to aldosterone 10-fold and increased by over 10-fold the mRNA levels of the late pathway enzyme aldosterone synthase. Sodium restriction had no effect on zona glomerulosa levels of 11 beta-hydroxylase mRNA. In two other normotensive rats, Dahl salt-resistant and Wistar Kyoto, sodium restriction again specifically increased aldosterone synthase mRNA without altering 11 beta-hydroxylase or cholesterol side-chain cleavage cytochrome P-450 mRNA levels. Thus, it appears that sodium restriction specifically increases late pathway aldosterone synthase mRNA levels, resulting in an increase in enzyme levels, followed by an increase in late pathway activity and an increase in aldosterone output. PMID- 8404676 TI - Decreased expression of protein kinase-C alpha, beta, and epsilon in soleus muscle of Zucker obese (fa/fa) rats. AB - Skeletal muscle is one of the first tissues to become insulin resistant in genetically obese rodents. The activation of protein kinase-C (PKC) in rat skeletal muscle is mediated by insulin stimulation of diacylglycerol (DAG) levels. Defects in the activation of PKC in the heart and liver of obese Zucker rats indicate that an abnormality in either stimulation of DAG or PKC occurs in obese tissues. DAG levels were significantly increased in soleus muscle from 15- to 19-week-old obese (fa/fa) Zucker rats. PKC activity was diminished in soleus muscle from fa/fa rats. Decreased levels of PKC alpha and -beta activity wer enoted after resolution of common PKC isozymes (Ca2+ and phospholipid dependent) by hydroxyapatite chromatography. Immunoreactivity of PKC-alpha, -beta, and epsilon also indicated that their levels are diminished in fa/fa soleus muscle by 70-90%. To determine at which level down-regulation occurs (i.e. gene expression or protein turnover), mRNA levels were examined by Northern blot analysis of total RNA. PKC alpha and -beta levels were diminished in Zucker obese soleus muscle compared to soleus from Zucker lean control (fa/-) animals, and PKC epsilon mRNA was not detected on the same blots. The transcript size for PKC beta mRNA in Zucker soleus muscle was unique. Both lean and obese Zucker muscle tissues expressed three transcripts that hybridized with the full-length PKC beta cDNA probes, with sizes ranging between 2.5-1.7 kilobases. Levels of all PKC beta transcripts were significantly decreased in obese Zucker tissues. Thus, levels of common PKC isozyme mRNA, protein, and enzyme activity in soleus muscle of the obese Zucker rat are decreased even though levels of the endogenous PKC activator DAG are elevated. The decreased levels of PKC may be related to the etiology of insulin resistance in skeletal muscle. PMID- 8404677 TI - Effects of N-methyl-D-aspartate receptor activation on cFos expression in luteinizing hormone-releasing hormone neurons in female rats. AB - N-Methyl-D,L-aspartic acid (NMA), an agonist of N-methyl-D-aspartate (NMDA) excitatory amino acid receptors, stimulates the secretion of LH by increasing the release of LHRH. During proestrus, LHRH neurons express cFos in association with the LH surge. To determine the involvement of NMDA receptors in the activation of LHRH neurons on proestrus, we treated animals with an NMDA receptor blocker, MK 801. Treatment with MK-801 (0.3 mg/kg, sc) at 1130 h blocked both the LH and PRL surges and cFos expression in LHRH neurons. These data suggest that NMDA receptors are involved in the regulation of LHRH neuronal activation during the LH surge. We then determined whether NMA treatment could restore LH secretion and cFos expression in LHRH neurons in animals whose endogenous proestrous LH surges were blocked with pentobarbital. In the pentobarbital-blocked rats, NMA failed to induce cFos expression in LHRH neurons and increase LH secretion, but it did result in an increase in PRL secretion. To determine if NMA treatment alone could induce cFos expression in LHRH neurons, diestrous rats were treated with NMA by either systemic (40 mg/kg BW; four injections, 10 min apart) or third ventricular (2 micrograms in 2 microliters; four injections, 10 min apart) injections. NMA administration (regardless of the route of administration) caused an increase in LH secretion and significant cFos expression in many regions of the brain, including sites where the LHRH perikarya are concentrated. However, neither systemic nor intraventricular administration of NMA induced cFos expression in LHRH neurons. Thus, even though NMA results in increased activity of LHRH neurons, as evidenced by increased LH secretion, NMDA receptor activation alone appears to be insufficient to induce cFos expression in the LHRH neurons. PMID- 8404678 TI - Local sympathetic denervation of white adipose tissue in rats induces preadipocyte proliferation without noticeable changes in metabolism. AB - The direct influence of the sympathetic nervous system on white adipose tissue was studied by performing a unilateral surgical denervation of the retroperitoneal fat pad in rats, the contralateral pad being used as a control. One week after surgery, the weight of the denervated pad was significantly higher than that of the intact pad. In vivo, glucose utilization was not altered by denervation. The expression of GLUT4 as well as the expression and activity of fatty acid synthase, lipoprotein lipase, and hormone-sensitive triglyceride lipase were similar in the two pads. Lipolysis in response to norepinephrine, determined in vitro, was not modified by denervation although the ratio between alpha 2- and beta-adrenergic receptors was changed. Denervation induced an increase in DNA content without change in the number of mature adipocytes. The expression of A2COL6/pOb24, a marker of the early step of adipocyte differentiation, was significantly enhanced in the denervated pad, suggesting an increased number of preadipocytes. This was confirmed by an increased cell number observed in the denervated fat pad 1 month after surgery. In conclusion, surgical denervation of the white fat pad does not alter the glucose and lipid metabolisms. By contrast, it accelerated adipocyte differentiation and led to the recruitment of new precursors. PMID- 8404679 TI - Up-regulation of beta 1-adrenoceptor messenger ribonucleic acid in the rat pineal gland: nocturnally, through a beta-adrenoceptor-linked mechanism, and in vitro, through a novel posttranscriptional mechanism activated by specific protein synthesis inhibitors. AB - The uniquely high concentration of beta 1-adrenoceptor mRNA (beta 1-mRNA) in the pineal gland provides a model for the regulation of GTP-binding protein (G protein)-linked receptor gene expression within a functioning endocrine gland. By Northern analysis it has been shown that a nocturnal up-regulation of beta 1-mRNA in the rat pineal results in a 2.6-fold increase in mRNA levels at the middark phase (2400 h) compared with those at the midlight phase (1200 h). This increase is blocked by administration of the beta-adrenoceptor antagonist propranolol before the onset of darkness. In vitro studies of beta 1-mRNA expression in organ cultured pineals has confirmed beta-adrenoceptor-linked up-regulation of beta 1 mRNA. Treatment of cultured pineals with the second messenger drugs forskolin and phorbol 12,13-dibutyrate produced mRNA responses that were again consistent with a primary role of beta-adrenoceptors in the up-regulation of beta 1-mRNA. Nuclear run-on analysis showed that the acute up-regulation of mRNA was mediated largely through a transcriptional mechanism. An additional novel mode of regulation of beta 1-mRNA was also identified; the protein synthesis inhibitors anisomycin and cycloheximide, but not puromycin and emetine, elicited an acute increase in beta 1-mRNA in cultured pineals that was not transcriptionally mediated. The mechanism underlying this mode of regulation is discussed with relation to control of the cellular immediate early gene c-fos. The facility to examine both physiological and in vitro changes in beta 1-mRNA expression in the pineal will provide further insight into the complexity that is apparent for the molecular regulation of G protein-coupled receptors. PMID- 8404680 TI - Tumor necrosis factor-alpha attenuation of luteinizing hormone-stimulated androstenedione production by ovarian theca-interstitial cells: inhibition at loci within the adenosine 3',5'-monophosphate-dependent signaling pathway. AB - Tumor necrosis factor-alpha (TNF) blocks LH-stimulated androstenedione production by immature rat theca-interstitial cells (TIC) in vitro. The mechanism for TNF inhibition of LH-induced androstenedione is unknown and was investigated. LH stimulation of androstenedione synthesis in TIC is mediated via a cAMP-dependent signaling pathway. LH-stimulated cAMP in TIC-conditioned medium was reduced in a biphasic manner by TNF at 1 and 48 h, but not at 4 and 24 h. To determine whether inhibition of cAMP resulted from TNF interference of LH binding, TIC were given TNF for 24 and 48 h, and LH binding was determined. TNF inhibited LH binding at 24 and 48 h. Scatchard analysis revealed a TNF-induced decrease in LH receptor number without altered affinity. TIC were given TNF and cAMP analogs [N6-benzoyl cAMP, 8-thiomethyl-cAMP, 8-(6-aminohexyl)amino-cAMP, and N6-2'-O-(Bu)2cAMP], which selectively activate cAMP-dependent protein kinase (PKA) type I and/or PKA type II, respectively. At 48 and 96 h, TNF blocked androstenedione production stimulated by all combinations of cAMP analogs; however, androstenedione synthesis recovered by 48 h after removal of TNF. Peak PKA activity in TIC was observed at 30 min in the presence of LH or cAMP analogs. LH- or cAMP analog directed PKA activity was inhibited after concomitant exposure to TNF; however, a 24-h pretreatment with TNF did not affect cAMP analog-stimulated PKA activity. The results indicate that in the modulation of steroidogenesis, TNF acts at multiple sites in the PKA pathway. First, TNF suppresses LH-stimulated cAMP production by TIC. Secondly, inhibition of cAMP may result from TNF attenuation of LH binding, and thirdly, TNF inhibits PKA activity of TIC and, thus, attenuates androstenedione production. PMID- 8404681 TI - Differential regulation of steroidogenic enzymes during differentiation optimizes testosterone production by adult rat Leydig cells. AB - The postnatal differentiation of rat Leydig cells may be subdivided into three stages based on morphology and steroid production. The purpose of this study was to clarify the developmental mechanisms underlying increased testosterone production by measuring steady state levels of the mRNAs for three steroidogenic enzymes in isolated Leydig cells at each stage of differentiation. These include Leydig cell progenitors on day 21, immature Leydig cells on day 35, and adult Leydig cells on day 90. The steroidogenic enzymes were 1) cholesterol side-chain cleavage enzyme (CSCC), 2) 17 alpha-hydroxylase (P450-17 alpha), and 3) 3 alpha hydroxysteroid dehydrogenase (3 alpha HSD). We report that levels of CSCC and P450-17 alpha mRNAs increase, whereas 3 alpha HSD mRNA levels decline during the course of Leydig cell differentiation. The levels of 3 alpha HSD mRNA were high in progenitor Leydig cells that appeared to contain little smooth endoplasmic reticulum and decreased in cells as smooth endoplasmic reticulum developed and other enzyme mRNAs increased. These observations suggest that the factors that regulate 3 alpha HSD mRNA levels are startlingly different from those that regulate the mRNA levels of CSCC and P450-17 alpha. We conclude that the progressive increase in the capacity of differentiating Leydig cells to produce testosterone can be explained in part by an increase in the activity of enzymes that synthesize testosterone (CSCC and P450-17 alpha) and a decrease in the activity of an enzyme that metabolizes testosterone and its precursors (3 alpha HSD). PMID- 8404682 TI - Estrogen and hydroxysteroid sulfotransferases in guinea pig adrenal cortex: cellular and subcellular distributions. AB - This report describes for the first time the cellular and subcellular localization of estrogen sulfotransferase (EST) as well as the subcellular localization of hydroxysteroid sulfotransferase (HST) in the mammalian adrenal cortex. A 34-kilodalton EST and two HSTs with 3 alpha- and 3 beta-hydroxysteroid substrate specificities (32 and 33 kilodaltons, respectively) were previously purified from guinea pig adrenal cortex and characterized. Western blots were used to establish that two antisera generated against EST were highly specific for EST, whereas three antisera generated against the HSTs were highly specific for the HSTs, but did not distinguish between the 3 alpha- and 3 beta HSTs. Light and electron microscopic immunoperoxidase labeling with these antisera revealed that the sulfotransferases were expressed only within the ACTH-responsive layers of the guinea pig adrenal cortex, with EST localized to zona fasciculata and zona reticularis cells, and the HSTs confined to the zona reticularis. No labeling was detected in the zona glomerulosa or adrenal medulla. EST was concentrated in cell nuclei; sparse labeling was distributed throughout the cytoplasm. HST labeling was intense in smooth endoplasmic reticulum of zona reticularis cells, but was absent from nuclei. Ovoid inclusions about 1-4 microns in diameter, with no limiting membrane, were observed in zona reticularis cells; these inclusions were strongly labeled for both EST and HSTs. No gender-specific differences in distribution or labeling intensity were apparent. The high concentration of EST immunoreactivity in nuclei suggests that EST may play a role in modulating the ability of active estrogens to regulate gene expression in ACTH-responsive cells. The distribution of HST labeling suggests that sulfonation of adrenocortical 3 hydroxysteroids takes place largely within smooth endoplasmic reticulum in the zona reticularis in adult guinea pigs. PMID- 8404683 TI - Stimulatory effect of follicle-stimulating hormone on basal and luteinizing hormone-stimulated testosterone secretions by the fetal rat testis in vitro. AB - The in vitro effect of FSH on testosterone secretion by the fetal rat testis was studied. Testes were cultured in the presence or absence of either commercial human (h) FSH (Metrodine; 200 mIU/ml) or recombinant hFSH (200 mIU/ml) for 3 days and with 100 ng/ml ovine LH during the last 4 h of culture. To avoid a stimulatory effect by the 0.4% LH that contaminates Metrodine, the cultures were performed in the presence of a monoclonal anti-hLH beta antibody and with a concentration of Metrodine that had no short term stimulatory effect on testosterone production by the fetal testes in vitro. Metrodine treatment had a positive long term effect on both basal and LH-stimulated testosterone secretion by fetal testes explanted on days 18.5, 20.5, and 22.5 postconception, which was abolished by the addition of a monoclonal anti-hFSH beta antibody. LH-free recombinant FSH also augmented basal and LH-stimulated testosterone secretion of testes explanted on days 13.5, 14.5, and 18.5 postconception. The positive effect of recombinant hFSH appeared during the second day of treatment with day 14.5 and 18.5 testes and on the third day of treatment with day 13.5 testes. As it is widely accepted that FSH receptors are exclusively localized on Sertoli cells, these results suggest that on or before day 15.5 of fetal life, 1) Sertoli cells are able to respond to FSH, 2) Sertoli cells can produce factors that are able to act on Leydig cell function, and 3) Leydig cells are sensitive to FSH-induced Sertoli cell factors. In conclusion, this study points out a potential paracrine control of fetal Leydig cell function and/or differentiation by fetal Sertoli cells as soon as fetal Leydig cells differentiate. PMID- 8404684 TI - Effect of cytochalasin-B on the metabolism of polyphosphoinositides in andrenocortical cells. AB - We have previously shown that microfilament-disrupting agents inhibit steroid secretion by frog adrenocortical cells. To determine the role of microfilaments in the process of corticosteroid production, we studied the effects of cytochalasin-B and chaetoglobosin-C on polyphosphoinositide metabolism in myo [3H]inositol-prelabeled frog interrenal (adrenal) slices. Immunocytochemical labeling of adrenocortical cells in primary culture with actin antiserum showed that cytochalasin-B (5 x 10(-5) M) induced a complete and reversible disruption of microfilaments, whereas chaetoglobosin-C, a cytochalasin analog that cannot interact with actin, did not modify the organization of the microfilament network. Cytochalasin-B caused a dramatic inhibition of corticosteroid release from perifused frog interrenal slices, whereas chaetoglobosin-C did not affect steroid secretion. Analysis of labeled inositol phosphates and phosphoinositides revealed that cytochalasin-B, but not chaetoglobosin-C, caused a significant increase in tritiated inositol content (+38%) and concurrently inhibited the formation of polyphosphoinositides (-48%). Cytochalasin-B reduced the production of phosphatidylinositol (-63%), phosphatidylinositol monophosphate (-46%), phosphatidylinositol bisphosphate (-46%), and lyso-phosphatidylinositol (-66%). Cytochalasin-B also blocked the stimulatory effect of angiotensin-II on the breakdown of phosphatidylinositol, phosphatidylinositol monophosphate, and phosphatidylinositol bisphosphate and the formation of lyso-phosphatidylinositol and inositol phosphates. The present results provide evidence of a role for microfilaments in polyphosphoinositide metabolism in adrenocortical cells. These data indicate that microfilaments are required for the incorporation of inositol into membrane phospholipids and are necessary for angiotensin-II-induced phospholipase activation. PMID- 8404685 TI - Expression and hormonal regulation of the CCAAT enhancer binding protein-alpha during differentiation of rat ovarian follicles. AB - Follicular development involves both proliferation and differentiation of thecal and granulosa cells. The process is regulated by gonadotropins and paracrine and autocrine factors, including steroid hormones, presumably by the induction of different genes at specific time points. In the present study, the expression and distribution of the CCAAT enhancer-binding protein-alpha (C/EBP alpha) were studied in immature ovaries and in ovaries in which follicular growth and development were initiated with PMSG, whereas ovulation and luteal formation were induced by the injection of hCG. Ovaries were collected before and at different time points after PMSG (0, 6, 24, and 48 h) and hCG (0.25, 1, 3, 10, and 24 h) treatment for analyses of the contents of C/EBP alpha mRNA and protein and the cell-specific immunohistochemical localization of the protein. C/EBP alpha mRNA increased to maximal levels 24 h after PMSG treatment. The effect was specific for the ovary, as C/EBP alpha mRNA in the uterus did not change. C/EBP alpha mRNA decreased 10 h after hCG treatment and increased again in newly formed corpora lutea. Immunohistochemistry and immunoblotting demonstrated a similar increase in C/EBP alpha during follicular development. To examine the involvement of specific hormones in the regulation of C/EBP alpha, hypophysectomized immature rats were injected sequentially with estradiol and FSH. This treatment resulted in a substantial increase in C/EBP alpha mRNA and protein. These results demonstrate that C/EBP alpha is hormonally regulated in the ovary and suggest a role for C/EBP alpha during differentiation of ovarian cells and follicular development. PMID- 8404686 TI - Murine macrophages express abundant insulin-like growth factor-I class I Ea and Eb transcripts. AB - Hemopoietic cells have been reported to synthesize insulin-like growth factor-I (IGF-I) messenger RNA (mRNA), but the relative contribution of specific cell lineages that express these transcripts remains unknown. Reverse transcription and amplification of complementary DNA (cDNA) by the polymerase chain reaction were used to characterize full-length IGF-I mRNA transcripts in murine hemopoietic cells. The identity of transcripts encoding the entire prepropeptide was confirmed by restriction endonuclease digestion, Southern blotting, cloning, and Sanger sequencing. Abundance of IGF-I mRNA transcripts was assessed both by Northern blotting and sensitive ribonuclease protection assays followed by quantification with Phosphor-Imager analysis. Whereas IGF-I cDNA transcripts could be detected in a variety of leukocytes after polymerase chain reaction amplification, IGF-I mRNA was negligible or nondetectable in T and B cell lines and in those tissues containing a predominance of these cell types (e.g. spleen and thymus) by Northern blotting and ribonuclease protection assays. In contrast, elicited peritoneal macrophages, a macrophage cell line, microglia, and bone marrow macrophages differentiated in vitro expressed abundant IGF-I mRNA transcripts, whereas neither a premyeloid cell line nor freshly isolated bone marrow cells expressed significant transcripts. The 5'-identity of macrophage IGF I transcripts was established using an exon 2-derived IGF-I cDNA probe. All protected transcripts were foreshortened, indicating transcript initiation exclusively within exon 1, characteristic of extra-hepatic IGF-I mRNA. However, at the 3'-end, both IGF-I Ea (lacking exon 5) and IGF-I Eb (containing exon 5) mRNA transcripts were evident, with the Eb product being detected at levels similar to those present in hepatic cellular RNA. A large molecular size (26 kilodaltons) prepro-IGF-I peptide was also detected in macrophage cell lysates by Western blotting. Collectively, our observations show that: 1) among hemopoietic cells, myeloid rather than lymphoid cells are the major source of IGF-I; 2) macrophage IGF-I mRNA consists of class I Ea and Eb transcripts; 3) these transcripts are translated into protein; and 4) expression of IGF-I is directly associated with differentiation of bone marrow macrophages. PMID- 8404687 TI - 5'-Heterogeneity of the mineralocorticoid receptor messenger ribonucleic acid: differential expression and regulation of splice variants within the rat hippocampus. AB - The mineralocorticoid receptor (MR) cDNA we previously isolated from the rat hippocampus differs from the clone isolated from the kidney at the 5' untranslated (5'UT) region. The kidney clone (alpha MR mRNA) and the hippocampal clone (beta MR mRNA) possess unique 5'UT sequences of 220 and 300 nucleotides, respectively, but share an invariant peptide-coding domain and appear to encode an identical MR protein. The two mRNA variants may represent tissue-specific forms of the MR or may be coexpressed in the rat hippocampus along with other 5'UT variants. Here, we report that three mRNA subtypes were found in the hippocampus; their relative abundance was as follows: alpha = beta >> gamma. The three mRNA variants were differentially distributed within the hippocampal subfields, with the alpha form being highly enriched in CA2, dentate, the fasciculum cinereum, and the indusium griseum, whereas beta and gamma forms were evenly distributed through CA1-4. Adrenalectomy selectively increased alpha MR mRNA content, but the changes were restricted to CA1, CA2, and CA3 regions. We conclude that multiple MR mRNAs are differentially expressed in the rat hippocampus. The expression of alpha MR mRNA is specifically increased during adrenalectomy, suggesting that the increase in total MR mRNA content documented previously arises from a substantial increase in a single MR variant that elevates the total MR mRNA content, with the apparent elevation reflecting the average of regulated and unregulated transcripts. It is suggested from our data that a complex mechanism involving transcription and translation regulates MR expression in the rat hippocampus. PMID- 8404688 TI - Molecular identification of binding sites for calcitonin gene-related peptide (CGRP) and islet amyloid polypeptide (IAPP) in mammalian lung: species variation and binding of truncated CGRP and IAPP. AB - We have investigated the binding of rat [125I]islet amyloid polypeptide (IAPP) and [125I]calcitonin gene-related peptide (CGRP) to lung membranes from the rat, rabbit, and bull and have characterized the mol wt (M(r)) of the binding site for each ligand by chemical crosslinking. Results imply the existence of three distinct types of binding site demonstrated by both [125I]CGRP and [125I]IAPP in each of the three species investigated. These were differentiated by the relative potencies of displacement by rat CGRP, human CGRP-(8-37), rat IAPP, and the rat IAPP fragments IAPP-(8-37), IAPP-(12-37), IAPP-(25-37), and IAPP-(28-37). High affinity binding sites were identified for [125I]CGRP [rat Ki, 0.119 +/- 0.027 nM (n = 6); rabbit Ki, 0.944 +/- 0.075 nM (n = 6); bull Ki, 0.20 +/- 0.031 nM (n = 6)], and CGRP-(8-37) was found to displace [125I]CGRP in all species [rat Ki, 6.63 +/- 0.91 nM (n = 6); rabbit Ki, 22.70 +/- 3.79 nM (n = 6); bovine Ki, 26.9 +/- 0.21 nM (n = 3)]. Compared to CGRP-(8-37), displacement by IAPP also showed varying affinities that were similar to that of CGRP-(8-37) (rat), lower than that of CGRP-(8-37) (rabbit), or higher than that of CGRP-(8-37) (bull). Truncation of IAPP caused large parallel decreases in its affinity for [125I]CGRP in the rabbit and bull by the loss of residues 1-8 (rabbit) and 1-12 (bull), but was not as pronounced in the rat. [125I]IAPP demonstrated high affinity binding in each species [rat Ki, 5.86 +/- 0.86 nM (n = 6); rabbit Ki, 18.72 +/- 2.90 nM (n = 6); bull Ki, 1.97 +/- 0.40 nM (n = 6)]. Truncation of IAPP caused a reduction of its affinity for [125I]IAPP in all species by the loss of residues 1 28. Chemical cross-linking analysis indicated binding of both ligands to sites of 64,000 M(r) in the rat and 50,500 and 51,000 M(r) in the rabbit and bull, respectively. In addition, [125I]IAPP bound to to a site of 100,000 M(r) in the rat. [125I]CGRP and [125I]IAPP binding were reduced in the presence of guanosine 5-o-(3-Thiotriphosphate) in all species, indicating an association with G proteins. This study implies the existence of CGRP/IAPP-binding sites in the lungs of these species that show varying and complex patterns of displacement by CGRP, IAPP, and their fragments. PMID- 8404689 TI - Differential expression of gap junction connexins in endocrine and exocrine glands. AB - We have investigated the expression of three gap junction proteins and their corresponding mRNAs by secretory cells of a variety of endocrine and exocrine rat glands. By immunostaining cryostat sections (indirect immunofluorescence) with antibodies against connexins (Cx) 26, 32, and 43 and by hybridizing total glandular RNA (Northern blot) with cRNAs for these proteins, we have found that several endocrine glands (pituitary, parathyroid, pancreatic islets, and adrenal) express Cx43, variable levels of Cx26, and no Cx32, whereas several exocrine glands (lacrimal gland, salivary glands, pancreas, prostate, and seminal vesicle) express high levels of Cx32 and variable levels of Cx26, but no Cx43. Thus, different sets of proteins comprise the gap junctions of endocrine and exocrine glands. Together with the findings that an endocrine gland (thyroid) that discharges secretory products extracellularly before releasing them in the vascular compartment expresses both Cx43 and Cx32 and that an exocrine gland (preputial gland) that has a pheromonal role expresses Cx43, these observations suggest that the differential expression of gap junction connexins may be required to specify the endocrine or exocrine differentiation of a secretory cell. PMID- 8404690 TI - A primary cell culture system of luteinizing hormone releasing hormone neurons derived from embryonic olfactory placode in the rhesus monkey. AB - The purpose of this study is to establish a primary LHRH cell culture system using embryonic olfactory placode and to examine whether LHRH cells derived from olfactory placode and the migratory pathway of LHRH neurons mature in vitro. Six monkey fetuses at the ages of E34-E36 were delivered surgically and the area including the olfactory placode (PL) and the areas that encompass the migratory pathway (MP) were dissected out. The tissues were cut into small pieces and plated on collagen- or poly-L-lysine-coated glass coverslips in medium M199. Cultures were maintained for up to 33 days and immunostained for LHRH, GnRH associated peptide, neurofilament protein, neuron-specific enolase, and glial fibrillary acidic protein. LHRH positive cells were also positive for neurofilament proteins neuron-specific enolase, and GnRH-associated peptides, but negative for glial fibrillary acidic protein. In the first week of culture, LHRH cells remained within the explants of PL, were rounded (average dimensions: 13.0 x 11.3 microns) and stained lightly. By the second week a number of LHRH cells (15.7 x 13.6 microns) with neurites started to migrate out from PL explants, whereas some still remained in the PL. By the third week a large number of LHRH cells (19.3 x 9.4 microns) had migrated out from the PL. They were fusiform in shape with clear nuclei and extended long varicose neurites up to 500 microns in length. A few "pioneer" LHRH cells appeared to lead the migration of 100-400 LHRH cells forming 1-3 major migratory paths. In contrast, LHRH cells from MP explants migrated out sooner than those from PL explants. LHRH cells from the ventral part of the MP, which is close to the PL, migrated out by 1-2 weeks and formed several migratory paths, whereas LHRH cells from the dorsal part of the MP, which is farther from the PL, were scattered widely around explants and their neurites were extended tortuously. Cultured LHRH cells released LHRH into the media and responded to challenge with high K+. The results indicate that 1) primary LHRH neurons can be obtained from the embryonic PL and their migratory pathway, 2) these neurons migrate and mature in culture and 3) they are accessible for cellular and molecular studies. PMID- 8404691 TI - Receptor-mediated stimulatory effect of IL-1 beta on hyaluronic acid and proteoglycan biosynthesis by cultured rat ovarian cells: role for heterologous cell-cell interactions. AB - An increasing body of information supports the possibility that intraovarian interleukin (IL)-1 may play an intermediary role in gonadotropin-triggered ovulation. To further evaluate this hypothesis, we have examined the effect of IL 1 beta on ovarian proteoglycan/glycosaminoglycan economy, an established corollary of the preovulatory cascade. Rat ovarian cells were metabolically labeled with [35S]sulfate and [3H]glucosamine for 48h in the absence or presence of IL-1 beta, with or without an IL-1 receptor antagonist (IL-1ra). At the conclusion of this treatment period, total 35S and 3H incorporation into cell associated and extracellular proteoglycan/glycosaminoglycan species was determined. Treatment of whole ovarian dispersates with IL-1 beta (10 ng/ml) produced substantial increments in the accumulation of extracellular macromolecular material [11.5-, 2.9- and 2.6-fold for hyaluronic acid (HA), heparan sulfate (HS) and dermatan sulfate (DS) proteoglycans, respectively]. In contrast, only modest increments (< or = 1.7-fold) were noted for IL-1 beta treated granulosa cells (GC), theca-interstitial cells (TC), or 4:1 co-cultures (GC/TC) thereof. Treatment of whole ovarian dispersates with IL-1 beta also resulted in significant (P < 0.001) increments in the cell-associated accumulation of both HA (6.0-fold increase) and DS proteoglycans (3.4-fold increase). However, the cell-associated accumulation of HS proteoglycan was not significantly affected by IL-1 beta regardless of the cellular preparation under study. The concurrent provision of IL-1ra (5 micrograms/ml) all but neutralized the IL-1 beta effect on HA biosynthesis thereby suggesting mediation by specific ovarian IL-1 receptor(s). Taken together, these observations suggest that treatment of ovarian cells with IL-1 beta results in an overall increase in macromolecular biosynthesis as well as in redistribution favoring extracellular HA and DS (but not HS) proteoglycans. Moreover, since whole ovarian dispersates proved more responsive to IL-1 than isolated cellular components thereof, the present observations suggest an obligatory requirement for heterologous cell-cell interaction without which optimal HA or proteoglycan biosynthesis may not be realized. These observations along with the demonstration of IL-1-mediated amplification of gonadotropin-triggered ovulation provide strong indirect support for the view that IL-1 may be the centerpiece of an intraovarian regulatory loop concerned with the promotion of key periovulatory events. PMID- 8404692 TI - Methimazole inhibits FRTL5 thyroid cell proliferation by inducing S-phase arrest of the cell cycle. AB - Previous studies, using tritiated thymidine uptake assays, had indicated a nil or stimulatory effect of methimazole (MMI) on thyroid cell proliferation. Whilst examining cell cycle kinetics of FRTL5 thyrocytes, we observed an inhibitory effect of MMI on thyroid cell proliferation. To further examine this observation, FRTL5 cells whilst in log phase proliferation were exposed to media containing either 6H or MMI in 6H. Cell number and cell cycle kinetics were examined using flow cytometric DNA analysis every 24 hrs for 96 hrs. We found that MMI inhibited cell proliferation (as assessed by cell number) throughout the experimental period. Cell cycle analysis revealed a persistent arrest of cells in S phase. Concomitantly, there was a fall in the proportion of cells in both G0G1 and G2M phases, in keeping with cell cycle arrest in S phase. Taken in isolation, the finding of a high proportion of cells in S phase would suggest stimulation of cell proliferation, consistent with the findings of previous studies which used tritiated thymidine uptake assays to assess cell proliferation. However, the absence of a concomitant increase in total cell number renders this argument invalid and argues for a specific effect of MMI on the cell cycle. This study demonstrates a hitherto unrecognised inhibitory action of MMI on FRTL5 thyroid cell proliferation which has implications in understanding the broader effects of MMI on thyroid cell physiology. Additionally, this study highlights the dangers of using tritiated thymidine uptake measures as the sole indicator of mitogenic activity. PMID- 8404693 TI - Increase in serum calcium after administration of PTH. PMID- 8404694 TI - Orthodontic closure and transplantation in the treatment of missing anterior teeth. An overview. AB - Orthodontic closure and autotransplantation of teeth are valid treatment alternatives for young individuals with missing anterior teeth. These alternatives have, however, different indications and the choice should be based upon a comprehensive evaluation and diagnosis of the individual. Problems and limitations should be emphasized before treatment in order for the patient to have realistic expectations. The treatment plan will ultimately result from an evaluation of short- and long-term biologic and esthetic considerations. Indications, treatment principles and key factors in the treatment procedure are outlined in this overview. If transplantation is to be part of the treatment plan, the donor tooth which has the best prognosis should be selected. PMID- 8404695 TI - Resin-retained bridges in the treatment of traumatized dentition. AB - Resin-retained bridges can be used as the definitive treatment, as well as for interim treatment. Compared with conventional fixed prostheses, there are some advantages which make this treatment especially useful in traumatized dentition. The ultra conservative approach makes the adhesive bridge a standard treatment option in cases of uncertain prognostic factors. However, in such specific situations as traumatized dentition some complicating factors may be present. These complicating factors must be recognized and eliminated to create situations in which this kind of restoration can be successful. Therapeutic and patient related factors are discussed using data from longitudinal clinical studies. Available clinical information indicates the anterior adhesive bridge to a 'permanent restoration'. However, a benefit-cost analysis is necessary to weigh the value of this restorations. PMID- 8404696 TI - Occlusal interferences in association with teeth left in the line of mandibular fractures. AB - In order to evaluate whether teeth left in the line of mandibular fracture can predispose to occlusal interferences, the contact relationships between the maxillary and mandibular teeth in 37 mandibular corpus fracture patients were examined on the average 41 months after the injury. Control group consisted of 27 healthy dental students. It was shown that occlusal interferences were encountered in 38% (16/42) of teeth in the line of mandibular fracture in 38% (16/42) of their anterior and in 32% (9/28) of their posterior ones. Similar examination in the control group revealed interferences in 28% (113/400) so the ones found in teeth in the line of mandibular fractures can be supposed to have been caused by the injury and/or its treatment. It is also concluded that whether or not the interferences found in these patients were caused by the injury or its treatment or were signs of individual range they should be corrected with occlusal adjustment to prevent further pathological changes in these teeth and in the function of the masticatory system. PMID- 8404697 TI - Traumatic injuries to permanent incisors. AB - The prevalence of traumatic injuries to permanent incisors and their distribution according to type and some clinical factors, were analysed in a total population of 2798 patients, aged 6-21 years, examined over a 5-year period in the Dental Clinic of the University of Verona, Italy. The material consisted of case histories and radiograms taken during the examination of injured teeth. The number of injured patients was 178 (131 males and 47 females), the number of injured incisors was 326. The prevalence of injuries was 7.3%. A very large number of dental injuries occurred to children aged between 6 and 13 years. The ratio of boys to girls was 2.7:1. Most frequent causes of injuries were falls and traffic accidents. Most injuries involved two teeth. About 80% of the teeth were maxillary central incisors. The most common type of injury was non complicated crown fracture. In 87 cases (48%) there was an associated injury to soft tissue. Alveolar fractures occurred in 22 cases (12%). Traumas involving periodontal ligament constituted nearly 40% of the total. This study noted the following points: 1) preventive education programs should be instituted in the region, directed at parents and school teachers, to inform them about the problems of dental trauma and to obtain cooperative attitude to treatment and good motivation in controls; 2) the necessity to unify the diagnostic and therapeutic protocol to provide reliable information in clinical investigations, to permit valid comparison with other studies and to improve the long-term prognosis of many cases. PMID- 8404698 TI - Effect of probe design on the suitability of laser Doppler flowmetry in vitality testing of human teeth. AB - The aim of this investigation was to study the influence of probe design on the signal output from the dental pulp in experiments with laser Doppler flowmetry (LDF). Eighteen patients 14 to 39 years of age were examined. Recordings were made from a maxillary and a mandibular central incisor and a maxillary canine with an infrared laser Doppler flowmeter. The radiographic appearance of the tested teeth was within normal limits and all teeth responded normally to electric pulp testing (EPT). Five configurations of probes were used. Each probe had 3 fibers arranged in a triangle. One fiber carried the laser light to the pulp tissue and 2 fibers carried the backscattered light to the detector giving the signal output. The diameter of each of the 3 fibers in the probe was 200 microns except in 1 probe where the diameter of the fibers was 125 microns. The distance between the 3 fibers in the triangular arrangement in each probe was 250, 500, 800, 1000, and 1500 microns. A special rubber base splint was used to hold the probe in place on the buccal surface of the tested teeth. The output signals from the LDF were fed into an analog printer and a lap top computer where all calculations were done. The probe with the largest separation of the fibers produced significantly higher output signals from the maxillary and mandibular incisors than the other probes. The same probe also produced significantly higher output signals than the smaller probes from the maxillary canine with the exception of the 200/800 probe.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404699 TI - Effects of systemic and topical nicotine on pulpal blood flow in dogs. AB - It has been suggested that nicotine exerts cardiovascular effects which are similar to stimulation of the sympathoadrenal system. If this observation is true, nicotine administration would decrease pulpal blood flow and possible alter the pulpal response to injury. The purpose of this study was to measure pulpal blood flow in dogs using the radiolabelled microsphere method following topical or systemic administration of nicotine for 28 days. Thirteen dogs were divided into three groups. Group one received topical nicotine (8 mg nicotine/kg/day) combined with orabase which was applied in two equal doses to the mandibular anterior gingiva. Group two received systemic nicotine (2.5 mg/kg/day) delivered by osmotic pumps implanted subcutaneously in the back of each animals' neck. Group three were controls, and these animals received either topical orabase twice daily applied to the mandibular anterior gingiva or saline via osmotic pumps. Results indicated pulpal blood flow increased from Day 0 to Day 28 in both nicotine treated groups. Group one (topical nicotine) exhibited a mean increase in blood flow of 21.8 ml/min/100 g, while group two exhibited a mean increase of 50.1 ml/min/100 g. Group three, the control animals, exhibited a mean decrease in pulpal blood flow of 22.1 ml/min/100 g over the 28-day interval. These changes were not statistically significant (p > 0.05). PMID- 8404700 TI - Influence of endodontic materials on the bonding of glass ionomer cement to dentin. AB - We assessed the bond strength of a glass ionomer cement to dentin that had been in contact with different materials. Flat dentin surfaces in freshly extracted human teeth were covered for 48 h with a 1 mm layer of a variety of materials that are used for temporary filling or root canal sealing. The products were mechanically removed and a glass ionomer cement cylindrical specimen bonded to the dentin surface. After 7-days immersion in 37 degrees C water the tensile bond strength was tested. The results were compared with those on dentin surfaces not in contact with any endodontic material. The statistical analysis showed that none of the materials used interfered with the bonding of the glass ionomer to dentin. PMID- 8404701 TI - Tactile sensitivity of three endodontic instrument handles. AB - Three currently available endodontic instrument handles, introduced over the past four decades, were investigated for tactility. Larger handles have been promoted to overcome sensitivity loss from wearing gloves. Smaller handles were preferred in this experiment. While the largest diameter handle appeared to give enhanced tactility, significance was not reached in any of the tests. PMID- 8404702 TI - Fracture of tooth by internal resorption: case report. AB - A case of extensive internal resorption of the left mandibular first premolar in a 24 year old man is reported. The case presented with slight discolouration and mobility of the tooth. On radiological examination extensive internal resorption and a fracture line at the cemento enamel region was observed, separating the crown and root. The case was treated endodontically after surgical removal of the fractured fragment of the crown. PMID- 8404703 TI - Internal resorption occurring after accidental extrusion of iodoform paste into the mandibular canal. AB - The precise etiology of internal resorption is unknown but it is generally believed that triggering factors are infection and persistent chronic pulpitis. A case is presented with internal resorption, which developed subsequent to extrusion of iodoform paste from the adjacent tooth into the mandibular canal. According to clinical and radiographic examinations made after 7 years, the tooth was clinically asymptomatic, vital and the internal resorption was arrested without treatment. PMID- 8404704 TI - Effect of midazolam on sphincter of Oddi motility. AB - Midazolam is a more recent benzodiazepine used for sedation during endoscopic procedures, including sphincter of Oddi (SO) manometry. However, the effect of this drug on the human SO has so far not been studied. In this paper we explored the effect of midazolam on human SO motility by means of endoscopic manometry. Twelve patients with suspected SO dysfunction were investigated. We found that in patients with normal manometry findings, midazolam had no effect on the sphincter motility. In contrast, in all patients with elevated SO pressure (SOP), as well as in one of the three patients with borderline SO tone, midazolam (2.5 mg i.v.) produced a relaxatory effect. Due to this effect the final readings were affected in three out of five patients, i.e. the abnormally elevated SOP decreased to a borderline level in two and in the remaining patient the borderline level in two and in the remaining patient the borderline SOP decreased to normal. In one other patient with a markedly elevated SOP the additional injection of 2.5 mg of midazolam caused a further decrease in SOP to a borderline level. We conclude that midazolam, due to its relaxatory effect on SO, appears to be a less suitable sedative for SO manometry. PMID- 8404705 TI - The effect of midazolam on motility of the sphincter of Oddi in human subjects. AB - Midazolam, a water-soluble benzodiazepine, has recently been introduced and found to be beneficial for sedation in upper GI endoscopy. Whereas it has been proven that diazepam does not have any effect on sphincter of Oddi (SO) motility in humans, no respective data exist on midazolam. On evaluation of the possible effects of midazolam on SO motility recording by means of endoscopic manometry, we found that midazolam had no effect on basal pressure (24.2 +/- 12.59 mm Hg before and 23.85 +/- 12.63 mm Hg after midazolam, p = 0.55), amplitude (39.75 +/- 22.62 versus 44.55 +/- 27.15, p = 0.097), duration (4.9 +/- 1.8 sec versus 5.05 +/- 1.7 sec, p = 0.614), and frequency of SO contractions (7.75 +/- 1.68 waves/min versus 7.15 +/- 1.92 waves/min, p = 0.083). These results suggest that in patients with normal manometric findings midazolam does not interfere with SO recording. PMID- 8404706 TI - The clinical need for sphincter of Oddi manometry in gastrointestinal endoscopy units. AB - The clinical need for sphincter of Oddi manometry (SOM) was investigated by retrospective analysis of 736 consecutive endoscopic retrograde cholangiopancreatography (ERCP) referrals (1985-89). During this period SOM was not performed in any unit in Greece including our own. Assuming a conservative or a more liberal policy in the utilization of SOM, a biliary and a pancreatic group of patients were established respectively, depending on the clinical presentation. ERCP was diagnostic in 168/194 (86.6%) of patients referred for post-cholecystectomy symptoms, but SOM was considered to be necessary to establish a diagnosis in the remaining 26 (13.4%) patients (biliary group). ERCP revealed pancreatic and/or biliary pathology in 46/69 (66.7%) patients, referred for symptoms attributed to pancreatitis, but 11/69 (15.9%) patients with pancreas divisum and 12/69 (17.4%) with acute recurrent idiopathic pancreatitis may have benefitted from SOM for planning endoscopic therapy (pancreatic group). Thus, only 5 patients from each group per 147 ERCP annual referrals were candidates for SOM. When taking into account that to run a SOM service the minimum annual number of investigations should be 50 and that the provision of ERCP in the UK is 50 per 100,000 of population per year, it is extrapolated that such a Gastrointestinal Endoscopy Unit should serve a population of 1.5 to 3 million. PMID- 8404707 TI - Endoscopic drainage of acute obstructive cholecystitis in patients with increased operative risk. AB - Emergency biliary surgery for acute obstructive cholecystitis in the elderly is associated with an increased hospital mortality. We therefore attempted to drain the obstructed gallbladder via the transpapillary route in 18 patients (mean age: 67 years) who had cystic duct obstruction on ERC and who were at an increased surgical risk. A cholecystonasal catheter was successfully introduced after a small EPT in sixteen of them (89%). This resulted in effective bile drainage, obviating the need for emergency surgery in all patients. No procedure-associated morbidity or mortality was found. Following clinical remission, elective treatment consisted of ESWL/direct stone dissolution (n = 10) or elective surgery (n = 3). Three patients received no further therapy. Our results show that endoscopic gallbladder drainage may be a valuable alternative to emergency surgery in high risk patients with acute obstructive cholecystitis. PMID- 8404708 TI - Carcinoid tumors of the rectum. PMID- 8404709 TI - Approach to the treatment of acute cholecystitis: open surgical, laparoscopic or endoscopic? PMID- 8404710 TI - How to sedate for endoscopic sphincter of Oddi manometry? PMID- 8404711 TI - Continuous 5-fluorouracil infusion causing acute gastric mucosal lesions. AB - A 59-year-old woman with advanced gastric cancer was treated with continuous 5 fluorouracil infusion for thirty-five days. Endoscopy of the upper gastrointestinal tract was performed due to continuing nausea seven days after the cessation of 5-fluorouracil infusion and revealed multiple erosions in the antrum which were absent before chemotherapy. Continuous 5-fluorouracil infusion was probably the cause of acute gastric mucosal lesions and should be included in the differential diagnosis of dyspepsia in 5-FU-treated patients. PMID- 8404712 TI - A carcinoid tumor of the rectum removed by strip biopsy. AB - In a 29-year-old male complaining of constipation, total colonoscopy revealed a 5 mm yellowish submucosal tumor at a distance of 6 cm from the anal ring. The histologic diagnosis of specimens obtained by biopsy was rectal carcinoid tumor and transanal ultrasonography revealed the tumor localization within the submucosal layer in the rectum. A strip biopsy was performed. Histopathological examination confirmed the diagnosis and ascertained complete excision. Rectal carcinoid tumors should be removed completely and strip biopsy may be a suitable procedure for removal of rectal submucosal tumors less than 10 mm in diameter. PMID- 8404713 TI - A post-sclerotherapy complication: intramural hematoma of the esophagus. PMID- 8404714 TI - Another severe complication of sclerotherapy for bleeding peptic ulcer. PMID- 8404715 TI - Endoscopic treatment of bleeding duodenal varices by bucrylate injection. PMID- 8404716 TI - Endoscopic treatment of a bleeding duodenal varix using N-butyl-2-cyanoacrylate. PMID- 8404717 TI - Are routine cleaning methods sufficient to remove Helicobacter pylori from endoscopic equipment? PMID- 8404718 TI - Transmission of Helicobacter pylori by endoscopy? PMID- 8404719 TI - Helicobacter pylori and intestinal metaplasia: possible implications in gastric carcinogenesis. PMID- 8404720 TI - Gastrointestinal metastases of an angiosarcoma. PMID- 8404721 TI - Endoscopic resection of two cases of gastric carcinoids. PMID- 8404722 TI - Small cell carcinoma of the esophagus in a cirrhotic patient. PMID- 8404723 TI - Duodenal ulcer perfusion during the healing process--an endoscopic laser-Doppler flowmetry study. PMID- 8404724 TI - The influence of the effectors of yeast pyruvate decarboxylase (PDC) on the conformation of the dimers and tetramers and their pH-dependent equilibrium. AB - The influence of effectors of yeast pyruvate decarboxylase, phosphate, pyruvamide, thiamin diphosphate and Mg++, on the pH-dependent equilibrium between dimers and tetramers was studied by synchrotron radiation X-ray solution scattering. Thiamin diphosphate and phosphate shift the equilibrium to higher pH values without altering the structure of the oligomers. Pyruvamide, a substrate analogue activator, induces a significant change in the structure of the tetramer. By eliminating radiation damage by addition of dithioerythrol to the buffers, the scattering curves could be measured accurately over a large angular range. They were expanded in terms of spherical harmonics to obtain the shapes of the dimers and tetramers with higher resolution than was hitherto possible. This also allowed us to position the dimers, which are centrosymmetric at low resolution, in the tetramers which have 222 symmetry. The results indicate that addition of pyruvamide results in a less compact tetramer owing to structural changes in the dimers and to their displacements. PMID- 8404725 TI - Induction of changes in the secondary structure of globular proteins by a hydrophobic surface. AB - Circular dichroism, ellipsometry and radiolabeling techniques were employed to study the induction of changes in the secondary structure of BSA, myoglobin and cytochrome C by a hydrophobic surface. The results showed that adsorbed protein molecules lose their ordered native structure in the initial stage of adsorption and the structure appears to be a random or disordered conformation. Protein molecules adsorbed in later stages adopt a more ordered secondary structure (alpha helix and beta structure). The changes of secondary structure of globular proteins induced by a hydrophobic surface can be explained by the steric interaction between adsorbed proteins as well as by hydrophobic interactions during the adsorption process. In addition, there is obviously an intermediate stage in which the protein molecules are mainly in the beta structure, indicating that for certain proteins, the beta structure may be a more stable secondary structure than alpha helix on the hydrophobic surface. PMID- 8404726 TI - Mathematical modelling of lipid transbilayer movement in the human erythrocyte plasma membrane. AB - A model is presented to simulate transverse lipid movement in the human erythrocyte membrane. The model is based on a system of differential equations describing the time-dependence of phospholipid redistribution and the steady state distribution between the inner and outer membrane monolayer. It takes into account several mechanisms of translocation: (i) ATP-dependent transport via the aminophospholipid translocase; (ii) protein-mediated facilitated and (iii) carrier independent transbilayer diffusion. A reasonable modelling of the known lipid asymmetry could only be achieved by introducing mechanism (iii). We have called this pathway the compensatory flux, which is proportional to the gradient of phospholipids between both membrane leaflets. Using realistic model parameters, the model allows the calculation of the transbilayer motion and distribution of endogenous phospholipids of the human erythrocyte membrane for several biologically relevant conditions. Moreover, the model can also be applied to experiments usually performed to assess phospholipid redistribution in biological membranes. Thus, it is possible to simulate transbilayer motion of exogenously added phospholipid analogues in erythrocyte membranes. Those experiments have been carried out here in parallel using spin labeled lipid analogues. The general application of this model to other membrane systems is outlined. PMID- 8404727 TI - 1992 William Lennox Lecture American Epilepsy Society annual meeting. The premise, the promise, and the problems with basic research in epilepsy. PMID- 8404728 TI - Burst suppression and impairment of neocortical ontogenesis: electroclinical and neuropathologic findings in two infants with early myoclonic encephalopathy. AB - We report the electroclinical and neuropathologic correlations in 2 children aged 2.5 months affected by early myoclonic encephalopathy characterized by epileptic seizures, erratic myoclonus, and an EEG pattern of burst suppression. Despite different etiologies, the neuropathologic findings showed similar abnormalities in both cases, with no substantial impairment of the myelination processes. Islands of matrix tissue scattered in the periventricular region and neurons aligned marginally in the bulbar olives were detected. The presence of numerous large spiny neurons dispersed in the white matter along the axons of the cortical gyri was the most striking finding. The neurons have been interpreted as abnormally persisting interstitial cells in 2.5-month-old children. These early generated neurons, normally present during neocortical histogenesis, are programmed to die near the end of gestation or soon after birth. The interstitial cells are regarded as a waiting compartment of afferent fibers during cortical development. Their persistence in our patients represents an anatomic condition for cortical disconnection providing a pathophysiologic basis to burst suppression phenomena. PMID- 8404729 TI - Epilepsy education in medical schools: report of the American Epilepsy Society Committee on Medical Student Education. AB - The education of medical students about epilepsy is often fragmentary and incomplete. Basic information regarding the classification of epileptic seizures, epilepsy syndromes, first aid for seizures, and practical use of antiepileptic drugs (AEDs) may not be covered in medical school curricula. In this report we describe reasons why the study of epilepsy provides a relatively unique opportunity to integrate concepts related to basic neuroscience and clinical neurology. Furthermore, we present a curricular framework that may prove useful for the education of medical students about epilepsy. PMID- 8404730 TI - Classification of epileptic seizures: a hospital-based study of 1,250 patients in a developing country. AB - The 1981 International Classification of Epileptic Seizures (ICES) was used to study the distribution of seizure types in 1,250 patients attending an Epilepsy Clinic in Sri Lanka. Based on seizure symptomatology 94.6% of the cases could be classified, and by adding the routine interictal EEG findings the percentage of classifiable seizures increased to 97%. Partial seizures (73.8% cases) were three times as common as generalized seizures (23.3% cases). Of the partial seizures, simple partial seizures (SPS) accounted for only 0.4% cases, and complex partial seizures (CPS) for 8.8%, whereas partial seizures secondarily generalized (PSGS) accounted for 64.6%. PSGS had simple onset in 12.5% and complex onset in 34.8% of cases. Myoclonic seizures were the commonest of the generalized seizures, accounting for 14.6% of all cases. Tonic-clonic seizures accounted for 7.4% of cases; absence seizures accounted for only 1.3%. The study showed the 1981 ICES to be relevant and applicable in a clinical setting with limited investigatory facilities. Difficulties encountered with regard to certain subcategories could be overcome with minor modifications which made the classification operative. Routine EEG confirmed the diagnosis in a significant number of cases but changed the diagnosis in only a few, confirming that a good standardized questionnaire is the key instrument for classifying epileptic seizures. PMID- 8404731 TI - Concordance of clinical forms of epilepsy in families with several affected members. Italian League Against Epilepsy Genetic Collaborative Group. AB - Evidence for genetic heterogeneity in epilepsy is strong. We evaluated the concordance of clinical forms in the same family in a series of families with several cases of idiopathic epilepsy, collected as part of the Study on the Genetics of Epilepsy of the Italian League against Epilepsy (LICE). The studied families had at least three members affected by an idiopathic form of epilepsy in one or more generations. Seventy-four families (with a total of 296 affected members) have been analyzed; two families had cases with benign neonatal familial convulsions (BNFC); in 25% of the remaining families all members were affected by the same clinical form, 13.9% had a prevalent clinical form with only one affected member with a different seizure type, 36.1% had two clinical forms, and 25% had three forms of epilepsy in the same family. There are no clinical differences in the form of epilepsy between the families concordant for one clinical form and families with two or three clinical forms of idiopathic epilepsies. The distribution of the clinical form in the affected relatives in our families showed the higher concordance with the proband in febrile convulsions (FC, 70.8%) and in epilepsy with generalized tonic-clonic seizures (EGTC, 63.0%). FC and EGTC were highly diffused in the affected relatives in the families with other forms of idiopathic epilepsy, above all in the more distantly related affected family members. In our families we observed a rare associated between childhood absence epilepsy (CAE) and juvenile myoclonic epilepsy (JME).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404732 TI - How common is catamenial epilepsy? AB - Forty women of childbearing age with refractory epilepsy were asked to record their seizures, the first and last days of their menstrual periods, and symptoms of premenstrual tension for 3 consecutive months. By defining catamenial epilepsy as the occurrence of at least 75% of seizures each month in the 10-day time frame, which included the 4 days preceding menstruation and the 6 days after its onset, only 5 women (12.5%) were identified who fulfilled the criterion. Nevertheless, after the study was completed, 31 (78%) of these patients claimed that most of their seizures occurred near the time of and were exacerbated by menstruation. The patients with catamenial epilepsy reported no more symptoms of premenstrual tension than did the rest of the group. Clustering outside the menstrual cycle was noted in 4 other patients. Catamenial epilepsy is an uncommon condition. Patient claims about frequency of seizures in relation to menstruation are not always accurate. A standard definition should be adopted because the diagnosis has implications for management. PMID- 8404733 TI - Seizures associated with propofol anesthesia. AB - Propofol is a new, fast-acting intravenous (i.v.) anesthetic. Involuntary movements or epileptic seizures have occurred during or after propofol-induced anesthesia in approximately 50 reported cases; a third of the patients have had epilepsy. We report 5 patients with seizures in association with propofol anesthesia. A female epileptic patient developed severe status epilepticus; the other patients with short-lasting seizures had no previous epilepsy. Although propofol has been used in treatment of patients of status epilepticus, the risk of precipitation of epileptic seizures warrants consideration especially when planning anesthesia for epileptic patients. PMID- 8404734 TI - Seizure-related injuries in multihandicapped patients with therapy-resistant epilepsy. AB - A prospective study on seizure-related injuries in Norway's two nursing homes for persons with epilepsy was conducted. Sixty-two multihandicapped patients with mostly difficult-to-treat epilepsy were assessed for 13 months: 6,889 seizures, 2,696 with ensuing falls, resulted in 80 injuries. The seizure-related injury risk was 1.2%. The most frequent injuries were mild soft tissue injuries with and without cuts. Six serious injuries were recorded: two leg fractures, one mandibular fracture, one neck of the femur fracture, one skull fracture, and one subdural hematoma. A 71-year-old woman with subdural hematoma died during operation for the hematoma. Seizure types most often causing injury were atonic and tonic-clonic seizures. Prophylactic measures can be taken. Because the seizure-induced injury risk was slight, we concluded that even persons with refractory epilepsy should be encouraged to lead active lives. PMID- 8404735 TI - Self-help epilepsy groups: an evaluation of effect on depression and schizophrenia. AB - The general prevalence of depression and schizophrenia was determined in a random outpatient population (n = 67) of refractory epileptic patients by Minnesota Multiphasic Personality inventory (MMPI) and Beck Depression Inventory (BDI). Of those patients 25% showed scores of moderate or severe depression. Self-help group intervention modifies significantly (p < 0.001) general mean depression rates, especially dystimia. Male and female differences emerged in our study for previous prevalence and pattern of recovery. PMID- 8404736 TI - Social adjustment in young adults with absence epilepsies. AB - A long-term follow-up of 58 young adults, aged 18-27 years, with persisting absence epilepsies since childhood or early adolescence, was performed to assess psychosocial outcome and the patients' own concept of their epilepsy. They were well adjusted in the areas of family status and employment, but had more unqualified jobs as compared with a reference group. They were also inclined to lead very regular lives in a way that led to social isolation. At least one of the following factors was considered by 74% of the group to have been affected by their epilepsy: schooling, occupation, routines of daily life, relations with friends, leisure time activities, and housing, this was independent of whether or not they had achieved seizure control. In treating absence epilepsies, it is important that one considers psychosocial aspects, even if a medically satisfying result with seizure control is obtained. PMID- 8404737 TI - Epilepsy and traffic safety. AB - We previously reported that drivers with epilepsy have somewhat higher age adjusted rates of traffic accidents and moving violations than do drivers without epilepsy. We attempted to identify medical and other factors contributing to this increase. Medical records of 241 drivers with a history of seizures, representing essentially all such persons from a contiguous seven ZIP postal code area served by the Marshfield Clinic were studied. This zip code refers to a defined geographic area around Marshfield where virtually the entire population receives its care at the Marshfield Clinic and for which we have accurate records. Information abstracted from medical charts was used to identify potential risk factors for traffic accidents and violations among these drivers. Careless driving violations, alcohol or drug violations, and accidents (especially injury accidents) occurred at higher rates and speeding violations occurred at lower rates for drivers with epilepsy. Young age, unmarried state, history of multiple seizures, and lack of antiepileptic drug (AED) treatment appear to be risk factors for accidents among drivers who had a history of seizures. Male sex, psychiatric disorders, alcohol abuse, and generalized seizures or complex partial seizures (CPS) were also suggestively associated with higher risk. For moving violations, young age, male sex, unmarried state, symptomatic etiology, and history of alcohol abuse contributed to increased risk. We conclude that drivers with epilepsy appear to have identifiable risk factors for traffic mishaps, especially accidents. PMID- 8404738 TI - Temporal lobe epilepsy in early childhood. AB - To explore the electroclinical features of temporal lobe epilepsy (TLE) in early childhood, we studied results of video-EEG and other tests of 14 children aged 16 months to 12 years selected by seizure-free outcome after temporal lobectomy. Four children had mesiotemporal sclerosis, 1 had cortical dysplasia, and 9 had low-grade temporal neoplasms. The children had complex partial seizures (CPS) with symptomatology similar to that of adults with TLE, including decreased responsiveness and automatisms. Automatisms tended to be simpler in the younger children, typically limited to lip smacking and fumbling hand gestures. Scalp/sphenoidal EEG showed anterior/inferior temporal interictal sharp waves and unilateral temporal seizure onset in the 4 children with mesiotemporal sclerosis and in the child with cortical dysplasia, but EEG findings in 9 children with low grade temporal tumors were complex, including multifocal interictal sharp waves or poorly localized or falsely lateralized EEG seizure onset. In children without tumors, video-EEG was critical to localization of the epileptogenic zone for resection, but in patients with tumors video-EEG was less localizing and its main value was to confirm that the reported behaviors were epileptic seizures with semiology typical of temporal lobe onset. PMID- 8404739 TI - Ictal 99mTc-HMPAO single photon emission computed tomography in children with temporal lobe epilepsy. AB - Seventeen ictal 99mTc-HMPAO single photon emission computed tomography (SPECT) studies were performed in 15 children with temporal lobe epilepsy (TLE) aged 7-14 years (mean 10.3 years). Ictal SPECT was informative in 16 of 17 (94%) studies in 14 of 15 (93%) children, showing unilateral temporal lobe hyperperfusion. In all 16 informative ictal SPECT studies, lateralization was concordant with ictal EEG, magnetic resonance imaging (MRI), and pathology. In 4 children, ictal SPECT provided additional localizing information that was not apparent from concurrent ictal EEG recording. Blinded interpretation of ictal SPECT studies by two independent investigators showed correct lateralization of the epileptic focus in every child. Results of visual analysis of ictal SPECT images were corroborated by quantitative analysis. Although interictal SPECT studies showed a degree of temporal lobe hypoperfusion in all children, in 9 of 15 hypoperfusion was either minimal, bilateral, contralateral, or associated with extratemporal hypoperfusion. In children with TLE, ictal SPECT provides reliable lateralizing information to corroborate or supplement that obtained from surface EEG and MRI. PMID- 8404740 TI - Temporal lobe epilepsy after prolonged febrile convulsions: excellent outcome after surgical treatment. AB - We studied 47 consecutive patients who underwent temporal resection for seizure control. Nineteen (40%) had febrile convulsions preceding onset of their habitual seizures. In 17 of 18 patients whose disease duration was known, the febrile convulsions were prolonged (mean 4 h). As compared with patients without preceding febrile convulsions, patients with antecedent febrile convulsions had a significantly higher prevalence of positive family history of febrile convulsions, an increased incidence of retrospectively identified gestational or perinatal complications, and no foreign tissue lesions. Pathologic studies showed gliosis and cell loss in mesiotemporal structures, usually moderate, in addition to usually mild gliosis in lateral temporal cortex. These patients had an excellent outcome after temporal resection: 84% were seizure-free, had residual auras only, or occasional convulsions with medication discontinuation. One patient (5%) had > 90% improvement. Two patients (11%) in whom the hippocampus was totally spared continued to have complex partial seizures: in both, seizures stopped after reoperation and hippocampal resection. Thus, 95% of these patients had an excellent result. Only 16% required invasive preoperative studies to confirm lateralization. These results were significantly better than those of the group without preceding febrile convulsions (p = 0.0013). PMID- 8404741 TI - Usefulness of unilateral interictal sharp waves of temporal lobe origin in prolonged video-EEG monitoring studies. AB - The value of EEG interictal epileptiform activity in predicting location of the seizure focus remains controversial. In 64 patients, scalp video-EEG monitoring studies showed one or two ipsilateral interictal foci in the temporal lobe. The site of these interictal foci correlated with location of the seizure focus recorded during prolonged video-electrocorticography (ECoG) with use of subdural grids placed under the mesiobasal temporal region and over the lateral temporal convexity. Our findings suggest that unilateral anterotemporal interictal foci can accurately predict location of seizure onset. This is also true in patients with two ipsilateral temporal interictal foci, provided that the dominant focus is localized in anterotemporal regions. We believe that in such patients invasive recordings are not warranted, but we caution against sole use of interictal epileptiform criteria for localization of the seizure focus. Correlation with clinical information, ictal EEG, neuropsychometric, and neuroimaging studies is required before performance of epilepsy surgery. PMID- 8404742 TI - New topographic mapping of temporal lobe seizures. AB - A unique topographic map has been developed based on EEG data of ictal events originating from the basal/mesiotemporal lobe regions. This technique involves a new mapping method of temporal lobe seizures as opposed to the interictal activity maps of most commercially available software. The map integrates data from sphenoidal electrodes as well as the standard 10-20 surface electrodes recorded with bipolar montages. A basal view is ideal for visualization of onset of temporal lobe ictal discharges recorded with chronic sphenoidal electrodes. We used the last 150 ictal events from 40 patients with basal/mediotemporal lobe epilepsy to develop this technique. Results indicate that a topographic view incorporating sphenoidal and scalp electrodes may provide a useful adjunct for interpretation of EEG recordings and a basis for comparison between and among patient groups for both ictal and interictal epileptic discharges. PMID- 8404743 TI - Effect of isoflurane (Forane) on intraoperative electrocorticogram. AB - Isoflurane, an inhalation agent often used for general anesthesia during craniotomy, has been reported to suppress spike activity in the intraoperative electrocorticogram (ECoG) during epilepsy surgery. We studied the effect of isoflurane concentrations of 0.25, 0.5, 0.75, 1, and 1.25% on the number of spike bursts per 5-min epochs in 15 patients undergoing ECoG during epilepsy surgery. N2O in O2 was maintained at 50% in 10 patients, at 60% in 2, and at 70% in 3. End tidal CO2 concentration was maintained in the hypocarbic range, and analgesia was maintained with the narcotic alfentanil in the range of 0.5-2 micrograms/min. The median number of spikes for each isoflurane concentration was 29 (range 3-107) at 0.25%, 27 (range 2-73) at 0.5%; 29 (range 5-90) at 0.75%, 33 (range 2-100) at 1%, and 40 (range 32-140) in 5 patients who tolerated 1.25% without occurrence of burst suppression pattern. No significant difference (Student's paired t test) was noted in the number of spikes for each isoflurane concentration. Therefore, if isoflurane concentrations are maintained between 0.25 and 1.25% or before burst suppression pattern occurs and N2O/O2 is maintained in the 50-70% range, isoflurane has no significant effect on spike activity. PMID- 8404744 TI - Partial seizures presenting as life-threatening apnea. AB - Apneic episodes, quite common in newborns, are considered rare after age 1 month, when gastroesophageal reflux, cardiac arrhythmias, idiopathic central apnea, and seizures become included in differential diagnosis. Determining the cause of apnea is important as treatment differs significantly and can be harmful; Caffeine given for presumed idiopathic central apnea is reported to have precipitated seizures in 2 patients with apneic seizures. Two cases of partial seizures presenting as apnea in infants were studied. Interictal EEG was normal in 1 and showed focal spikes in the other. Video EEG monitoring (16 channel) showed focal ictal discharge originating from temporal areas clearly preceding onset of apnea in both patients. Because therapeutic options are sometimes diametrically opposite and interictal EEGs are particularly unreliable for diagnosis, we recommend video-EEG monitoring if there is any doubt about the diagnosis before starting treatment in patients with apneic episodes. PMID- 8404745 TI - Corpus callosotomy for intractable epilepsy: seizure outcome and prognostic factors. AB - We reviewed the outcome of corpus callosal section in 64 adult and pediatric patients to identify factors associated with a good outcome: 48% of patients had a favorable outcome for overall seizure frequency. Improvement was noted in several seizure types and was most likely for drop attacks, particularly in the setting of a unilateral focal cerebral lesion or a true generalized epilepsy of Lennox-Gastaut type. Poor outcomes for drop attacks were more likely if there was associated severe intellectual handicap or bilateral independent spikes on interictal EEG. Complex partial seizures (CPS), most commonly of frontal lobe origin, also responded favorably. The complications of callosal section were usually mild and transient. New focal seizures occurred in only 2 patients and were not as frequent or disabling as preoperative seizures types. A worthwhile improvement in seizure outcome was achieved by completion of the callosotomy in 6 of 10 patients with unsatisfactory results from anterior callosotomy. PMID- 8404746 TI - Corpus callosotomy in treatment of medically resistant epilepsy: preliminary results in a pediatric population. AB - We report the results of 34 patients who underwent corpus callosotomy between 1986 and 1989 with 28-65 months of postoperative follow-up (mean 42 months). Thirty-two patients had mental retardation and 26 had significant behavioral problems. Thirteen patients had total section, 8 had subtotal section with preservation of the posterior half of the splenium, and 13 had section of the anterior two thirds of the callosum. Satisfactory seizure control was achieved in 25 patients (73.5%) Atonic seizures, followed by tonic seizures, generalized tonic-clonic seizures (GTCs), and atypical absence seizures were most improved. Myoclonic and complex partial seizures (CPS) did not improve significantly. No deterioration in seizure status was observed postoperatively. Two patients developed previously unobserved simple seizures and CPS postoperatively, but they were not as disabling as the preoperative seizures. Among the patients with behavioral problems, 81% had significant decrease in aggressiveness, hyperactivity, and/or attention deficit. Patients who underwent total section had interhemispheric disconnection symptoms that improved progressively and did not interfere with daily life. Decreased speech output, dysarthria, and gait dyspraxia occurred after total callosal section and persisted in 5 of the 13 patients. Patients who underwent anterior two thirds or subtotal sections did not have such symptoms. Early postoperative complications consisted of aseptic ventriculitis (5), subdural hematoma (1), and wound infection (4) and resolved without sequelae. PMID- 8404747 TI - Suppression of interictal spikes and seizures by stimulation of the vagus nerve. AB - The effects of electrical stimulation of the vagus nerve, a proposed treatment for patients with intractable epilepsy, on focal interictal spikes produced by penicillin and EEG secondarily generalized seizures induced by pentylenetetrazol were assessed in rats. Interictal spike frequency was reduced by 33% during 20 s of stimulation (p < 0.001) and remained low for < or = 3 min. Amplitude of residual spikes was also decreased. Cardiac and respiratory rates were suppressed. Cooling the nerve proximal to the point of stimulation abolished the EEG and respiratory effects. A similar reduction in spike frequency of 39% was obtained by heating the animals' tail (p < 0.01). Vagal stimulation at onset of seizures reduced mean seizure duration from 30.2 +/- 15.7 s without stimulation to 5.0 +/- 1.8 s (p < 0.01). Only the EEG equivalent of the clonic phase of the seizure was affected. These findings suggest that vagus nerve stimulation can be a potent but nonspecific method to reduce cortical epileptiform activity, probably through an indirect effect mediated by the reticular activating system. PMID- 8404748 TI - Motor and sensory impairments in children with intractable epilepsy. AB - During a 3-year period (1988-1991), 72 children with severe intractable epilepsy were studied. A standardized protocol for assessment of motor and sensory function was designed for school age children. Function was quantified on a 4 point scale on 47 items, including gross motor function, balance, coordination, strength, range of motion (ROM), velocity, fine motor function, sensation, perception, and neurologic tests. Classification of handicaps according to World Health Organization (WHO) definitions was performed. Videotape documentation completed the assessment. Evaluation of treatment services showed that provision of rehabilitation services had been insufficient and provided only for children with additional major movement disorders, mainly cerebral palsy (CP) cases. To minimize the handicap in children with severe epilepsy, it is essential to clarify the total sensorimotor impairment pattern, including balance, coordination, and perceptual capacity. Impairments in these functions are, as shown in this study, frequent and exist independent of major disabilities such as mental retardation or cerebral palsy. When several neuroimpairments were identified, a multiplicative rather than an additive effect on the total handicap was evident. PMID- 8404749 TI - Remission of seizures and predictors of intractability in long-term follow-up. AB - In an incidence cohort, remission and relapse rates and determinants were studied in 178 patients followed long-term. A comparative study of predictive factors was performed in 40 patients with histories of antiepileptic (AED)-drug-refractory epileptic seizures in the last 10 years of follow-up and compared with the other 138 cohort subjects. The two groups were cross-tabulated with 353 variables of family history, obstetric, developmental and seizure histories, and current medical and social status. Multivariate analyses were applied for control of confounding. Defined or probable remote symptomatic etiology of seizures, abnormal neurologic development/status, high initial seizure frequency, occurrence of status epilepticus, and poor short-term effects of AED therapy were significantly associated with long-term AED refractoriness. On logistic regression analyses, poor short-term outcome of AED therapy [odds ratio (OR) 3.6; 95% confidence interval (CI) 1.2-10.4], occurrence of status epilepticus (OR 11.4; 95% CI 3.2-41.0), high initial seizure frequency (OR 4.6; 95% CI 1.1-19.3), and remote symptomatic seizure etiology (OR 2.9; 95% CI 1.1-8.2) remained the only independent predictors of seizure intractability. These factors enable early assessment of need for epilepsy surgery. PMID- 8404750 TI - Adjuvant vigabatrin in refractory epilepsy: a ceiling to effective dosage in individual patients? AB - A double-blind, randomized, cross-over study of additional vigabatrin (gamma vinyl-GABA, VGB, 1.0 g twice daily for 6 weeks, followed by 1.5 g twice daily for 6 weeks) and matched placebo was undertaken in 24 patients with refractory epilepsy. Nineteen completed the trial satisfactorily. Fewer seizure days were reported during VGB treatment [placebo 41, VGB 23, p < 0.05, 95% confidence interval (CI) -1.5 to -14]. An overall reduction in median seizure numbers failed to reach statistical significance (n = 19; placebo 52, VGB 32, NS, 95% CI -18 to +24). Subgroup analysis, however, showed a significant reduction in partial seizures (n = 17) with 2 g VGB daily (placebo 22, VGB 13, p < 0.05, 95% CI -0.5 to -16.5), but not with higher dosage (placebo 28, VGB 22, NS, 95% CI -18 to +11). A deterioration in control of partial seizures as compared with the equivalent placebo phase was observed when patients were changed from 2 to 3 g/day VGB (2 g VGB 13, 3 g VGB 22, p = 0.05, 95% CI 0 to +20). Loss of efficacy was noted in 3 patients, and seizure control worsened slightly in 5 others. One previously resistant patient developed a therapeutic response, and 2 other patients reported an additional useful reduction in seizures. In the remaining 8 patients, seizure frequency did not change. VGB did not appear to benefit tonic clonic seizures. Serum VGB concentrations were higher during treatment with 3 g (15.5 +/- 8.9 mg/L) daily than with 2 g (13.5 +/- 11.2 mg/L). No important alterations were noted in the concentrations of concomitantly administered antiepileptic drugs (AEDs) throughout the trial. VGB is useful adjuvant therapy for treatment of partial seizures. There may be a ceiling to effective dosage. This demands individual dose titration for each patient. PMID- 8404751 TI - Increasing plasma concentration tolerability study of flunarizine in comedicated epileptic patients. AB - Twelve patients with intractable partial seizures [4 receiving carbamazepine (CBZ), 4 phenytoin (PHT), and 4 both] entered a study of the tolerability of flunarizine (FNR) at specified plasma concentrations. After an 8-week baseline period, a single-dose pharmacokinetic study was performed for each patient to calculate a loading dose and maintenance dosage necessary to achieve a target plasma FNR concentration of 30 ng/ml. The first 8 patients received the loading dose (as divided doses) during a 1-week hospitalization and the maintenance dosage for the ensuing 8 weeks. These patients proceeded to treatment periods with target concentrations of 60 and then 120 ng/ml, using doses based on an assumed linear relation between dose and plasma concentration. The last 4 patients were studied only at the 120- ng/ml target level. Results indicated that this procedure successfully approximated target levels of 30 and 60 ng/ml, but observed concentrations in the last period exceeded the 120-ng/ml target level and continued to increase with time, often necessitating a dosage reduction owing to intolerability. Calculated doses for a given target concentration varied by a factor of 12. The most frequently reported adverse experiences were sedation and increased fatigue; reports of dizziness, headache, and lethargy were also common. Based on this study, a target concentration of at least 60 but < 120 ng/ml is recommended for a controlled clinical trial of the antiepileptic efficacy of FNR. PMID- 8404752 TI - Periodontal condition of epileptic adults treated long-term with phenytoin or carbamazepine. AB - The periodontal condition of 40 adult epileptic subjects (mean age 51 years) receiving long-term therapy (mean 18 years) with phenytoin (PHT) or carbamazepine (CBZ) was studied. The subjects completed a questionnaire and underwent clinical and radiologic examination. Patients receiving PHT exhibited the same level of alveolar bone loss as those receiving CBZ. Patients receiving PHT exhibited more units with gingival overgrowth, reflected by the significantly higher number of gingival units with increased probing depth (p < 0.05). The results indicate that long-term PHT does not result in increased risk for alveolar bone loss as compared with CBZ. PMID- 8404753 TI - Carbamazepine-induced tics. AB - A variety of movement disorders are known to occur in association with carbamazepine (CBZ) therapy in adults and children, but development of tics has been described infrequently and only in patients with underlying Tourette's syndrome or other movement disorders. We report 3 children with epilepsy who developed facial motor tics after initiation of CBZ for complex partial seizures. All 3 had documented CBZ blood levels in the therapeutic range at the time, and none had other symptoms or signs of clinical intoxication. Neurologic examinations were normal in 2 and showed developmental delay of expressive language in the third. Brain imaging was normal in all. After development of the tics in 2, CBZ was continued at the same or higher dose, and the tics abated and then ceased spontaneously < or = 6 months. In the third child, the tics ceased after CBZ discontinuation. These cases demonstrate that CBZ can induce simple motor tics in children. These idiosyncratic reactions may be transient and do not always necessitate drug discontinuation. PMID- 8404754 TI - The balance of ecological destruction and repair. PMID- 8404755 TI - Gender-related behavior in women exposed prenatally to diethylstilbestrol. AB - Accumulating evidence in experimental animals over the past three decades suggests that mammalian brain development and differentiation of the central nervous system are influenced by perinatal exposure to sex hormones. Hence, changes in human behavioral patterns may be associated with prenatal exposure to estrogenic substances such as diethylstilbestrol (DES). This paper reviews relevant studies from a series of laboratories and finds that no clear-cut differences can be demonstrated to date between unexposed and DES-exposed women in gender-related behavior, although the physical and psychological impact of the problems associated with exposure to DES are well documented. If both prenatal and postnatal influences such as social, economic, and environmental factors are taken into consideration, individual variation is more apparent than differences in gender-related behavior between unexposed and DES-exposed women. In summary, gender-related behavior is determined by a complex array of interacting factors, and prenatal influences are only one of many developmental events. More studies are needed using larger populations with carefully controlled selection criteria to suggest a direct role of prenatal DES exposure on subsequent gender-related behavior. PMID- 8404756 TI - Orphan receptors. PMID- 8404757 TI - Toxicity tests in animals: historical perspectives and new opportunities. PMID- 8404758 TI - NAFTA prompts health concerns across the borders. PMID- 8404759 TI - The fungus among us: use of white rot fungi to biodegrade environmental pollutants. PMID- 8404760 TI - Neurogenic inflammation and sensitivity to environmental chemicals. AB - Neurogenic inflammation as a pathway distinct from antigen-driven, immune mediated inflammation may play a pivotal role in understanding a broad class of environmental health problems resulting from chemical exposures. Recent progress in understanding the mediators, triggers, and regulation of neurogenic inflammation is reviewed. Evidence for and speculations about a role for neurogenic inflammation in established disorders such as asthma, rhinitis, contact dermatitis, migraine headache, and rheumatoid arthritis are presented. The sick building syndrome and multiple chemical sensitivity syndrome have been defined as clinical entities in which exposure to chemical inhalants gives rise to disease. Current data on the existence of chemical irritant receptors in the airway and skin are discussed; neurogenic inflammation arising from stimulation of chemical irritant receptors is a possible model to explain many of the aspects of chemical sensitivities. PMID- 8404761 TI - Effectiveness of flushing on reducing lead and copper levels in school drinking water. AB - Samples from drinking water fountains in 50 schools in New Jersey were collected at specific times during a typical school day and analyzed for lead, copper, pH, alkalinity, and hardness. First-draw lead and copper levels (medians 0.010 mg/l and 0.26 mg/l, respectively) decreased significantly after 10 min of flushing in the morning (medians 0.005 mg/l lead and 0.068 mg/l copper), but levels increased significantly by lunchtime (medians 0.007 mg/l lead and 0.12 mg/l copper) after normal use of fountains in the morning by students. Corrosive water, as defined by the aggressive index, contained significantly higher levels of lead and copper (medians 0.012 mg/l and 0.605 mg/l, respectively) than noncorrosive water (medians 0.005 mg/l and 0.03 mg/l, respectively). PMID- 8404762 TI - Enhanced response to ozone exposure during the follicular phase of the menstrual cycle. AB - Exposure to ozone (O3), a toxic component of photochemical smog, results in significant airway inflammation, respiratory discomfort, and pulmonary function impairment. These effects can be reduced via pretreatment with anti-inflammatory agents. Progesterone, a gonadal steroid, is known to reduce general inflammation in the uterine endometrium. However, it is not known whether fluctuations in blood levels of progesterone, which are experienced during the normal female menstrual cycle, could alter O3 inflammatory-induced pulmonary responses. In this study, we tested the hypothesis that young, adult females are more responsive to O3 inhalation with respect to pulmonary function impairment during their follicular (F) menstrual phase when progesterone levels are lowest than during their mid-luteal (ML) phase when progesterone levels are highest. Nine subjects with normal ovarian function were exposed in random order for 1 hr each to filtered air and to 0.30 ppm O3 in their F and ML menstrual phases. Ozone responsiveness was measured by percent change in pulmonary function from pre- to postexposure. Significant gas concentration effects (filtered air versus O3) were observed for forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV1), and forced expiratory flow between 25 and 75% of FVC (FEF25-75; p < .05). More importantly, the pulmonary function flow rates, FEV1 and FEF25-75, showed a significant menstrual phase and gas concentration interaction effect, with larger decrements observed in the F menstrual phase when progesterone concentrations were significantly lower. We conclude that young, adult females appear to be more responsive to acute O3 exposure during the F phase than during the ML phase of their menstrual cycles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404763 TI - Molecular modeling studies suggest that zinc ions inhibit HIV-1 protease by binding at catalytic aspartates. AB - Human immunodeficiency virus type 1 protease is inhibited in vitro by zinc ions at neutral pH. The binding site of these ions is not known; however, experimental data suggest that binding may occur in the active site. To examine the possibility of zinc binding in the active site, molecular dynamics simulations in the presence and absence of zinc have been carried out to 200 psec. The results are compared with the 2.8-A crystallographic structures of a synthetic HIV-1 protease, and a zinc binding site at the catalytic aspartate residues (Asp-25, Asp-25') is proposed. Molecular dynamics simulations show that the zinc ion remains stably bound in this region, coordinating the carboxylate side chains of both aspartate residues. Interaction with zinc does not disrupt the dimeric structure of the protein or significantly alter the structure of the active site. These data are consistent with experimental studies of HIV-1 protease inhibition by zinc and give strong evidence that this is the binding site that leads to inactivation. PMID- 8404764 TI - Effect of intermittent exposure to sunlight on melanoma risk among indoor workers and sun-sensitive individuals. AB - Intermittent exposure to sunlight is considered to be an important risk factor for melanoma, but the associations reported in most case-control studies are surprisingly weak. The aim of this study was to evaluate whether the incorporation of a subject's background exposure to the sun and pigmentation characteristics (which are assumed to influence a person's susceptibility to sunlight exposure) could produce stronger associations between sunlight exposure and the risk for melanoma. A population-based case-control study was performed in the mid-eastern part of the Netherlands. The study group comprised 141 patients with a histologically verified melanoma and 183 controls with other malignancies who were registered by the same cancer registry. Patients with a lentigo maligna melanoma or an acrolentiginous melanoma were excluded. Information was collected by interviews and physical examination. We categorized subjects as indoor or outdoor workers on the basis of occupational exposure to the sun. Pigmentation characteristics, which are known to be risk indicators for cutaneous melanoma, were summarized as one sun sensitivity score. We used this score to distinguish between sun-sensitive and sun-resistant persons. The odds ratios associated with sunbathing, vacations spent in sunny countries, and sunburns were higher among the indoor workers than among the outdoor workers. After stratification by the sun sensitivity score, the effect of sunbathing, participating in water sports (swimming excluded), vacations to sunny countries, and a history of sunburn was largest for the sun-sensitive persons. The data show a general trend toward higher relative risks among indoor workers and sun-sensitive individuals. The results of this study support the intermittent sunlight hypothesis. PMID- 8404765 TI - Human experimental MIBK exposure: effects on heart rate, performance, and symptoms. AB - Heart rate, performance, and symptoms were studied in six female and six male volunteers, aged 19 to 47 years, during experimental 2-hr exposures to 10 and to 200 mg/m3 of methyl isobutyl ketone (MIBK). No effects from exposure on performance of a reaction time task or an arithmetic test could be demonstrated, and no consistent effects on heart rate were found. Subjects reported significantly more symptoms from the central nervous system, e.g., fatigue, due to the exposure. There was also an indication of an increase in ratings of irritation to the airways. A reduction of the threshold limit value (TLV) of 205 mg/m3 for MIBK exposure presently indicated by the American Conference of Governmental Industrial Hygienists, is therefore recommended. PMID- 8404767 TI - A cross-sectional survey of neurobehavioral effects of chronic solvent exposure on workers in a paint manufacturing plant. AB - Neurobehavioral impairments in 84 workers with long-term exposure to organic solvents in a paint manufacturing plant were examined cross sectionally. The World Health Organization (WHO) Neurobehavioral Core Test Battery, the NES-2 computerized battery, and four additional South African tests were used. Exposure to solvents was determined by using company industrial hygiene data as well as from an industrial hygiene survey of current total solvent levels in air. Indexes for cumulative exposure and average lifetime intensity of exposure were calculated for groups of homogeneously exposed workers in each department. Exposure levels were below the AGGIH threshold limit values. Multiple linear regression revealed that education level, age, and alcohol consumption were strong predictors for several neurobehavioral test scores. After adjusting for potential confounding from this source, average lifetime intensity of solvent exposure was the most significant predictor of the NES-2 Continuous performance test (measuring sustained visual attention) score of the WHO Digit span backward test score (measuring attention span and double tracking). Pursuit aiming (measuring fine visuomotor tracking speed) was significantly associated with the cumulative exposure index, possibly indicating early neurotoxic effects. PMID- 8404768 TI - Determination of evoked potentials in occupational and environmental medicine: a review. AB - The measurement of cerebral evoked and event-related potentials is a promising technique for assessment of subclinical neurotoxicity and has recently been introduced into occupational and environmental medicine. Evoked potentials consist of somatosensory, visual, and auditory evoked potentials, and event related potentials include the P300 potential. Measurement of these potentials can localize central nervous system impairments caused by exposure to a wide variety of hazardous factors in the workplace and the general environment. This paper is intended to provide an overview of research utilizing these potentials to evaluate the effects of work-related factors. The available data indicate that these potentials are sensitive and reliable methods that are easily standardized and practical to apply in the field setting. Researchers should note, however, that several covariates such as age, skin or body temperature, height, alcohol ingestion, and intelligence can influence assessment of these cerebral potentials in clinical and epidemiologic studies. PMID- 8404766 TI - The effects of ethanol exposure on radial arm maze learning and behavior of offspring rats. AB - The effects of maternal drinking on offspring have been studied epidemiologically, in human beings, and experimentally, in rats. The physical growth of offspring of female alcoholic rats, including histological growth of brain, lung, thymus gland, liver, and kidney, was previously reported by us. In the present study, we observed the effect of ethanol intake by the mother rat on learning ability and behavior of offspring rats using an eight radical arm maze. At the same time histological observations of the cerebrum were carried out. The mother rat was exposed to ethanol from a young age to delivery (P-DEL) or to weaning (P-NURS). After weaning, the offspring was exposed to ethanol until the tests began (P-WEAN). Experimental groups, classified by length of ethanol exposure, as mentioned above, disclosed the following: (1) Number of trials required for fulfilling learning criterion was significantly large in P-DEL and P NURS rat groups relative to the controls; that is, P-DEL and P-NURS rats were slow in learning. (2) Numbers of rats which did not fulfill the learning criterion were: Group P-DEL, one male of eight; Group P-NURS, three males of seven. The behavior of the rats in Group P-WEAN differed from those in other groups; while they were receiving acclimation training, they were, unlike ordinary rats, not watchful of the device, slow to find the feed, and indifferent. They seemed to lack carefulness and sometimes failed to eat the feed even though they succeeded in selecting correct arms. Their motion was abrupt and they ran at extraordinarily high speeds. (3) In the observations of correct choices in the first eight choices, groups P-DEL and P-NURS showed significantly low values. This suggested the lowering of their learning ability. (4) In the observations of continuous correct choices, Group P-DEL showed a significantly low value. This suggested the rats did not learn thoroughly enough to retain their acquisition long. (5) Body weight, learning ability, and hippocampal neurons were affected by ethanol exposure more severely in Group P-NURS than in Group P-DEL. An even more severe effect was observed in Group P-WEAN. PMID- 8404769 TI - Work stress in Japanese computer engineers: effects of computer work or bioeducational factors. AB - To examine whether computer work and bioeducational factors (age and school career) have significant effects on work stress in computer engineers in Japan, we administered a stress questionnaire to 764 male computer engineers and 211 male office workers in a computer-manufacturing factory in Tokyo. Four scales of perceived psychological stress at work examined were work overload, poor human relationships at work, unsuitable job, and competition-dismissal anxiety. The results of the three-way analysis of variance, in which age (20-29, 30-39, and 40 49 years), school career (high school and university graduates), and computer work (computer engineers and office workers) were three variation factors, indicated that: (1) there were no significant differences in all scores of work stress between computer engineers and office workers (P > 0.05); (2) scores for unsuitable job and poor human relationships at work were significantly higher in high school graduates than in university graduates (P < 0.05); and (3) there were significant age differences in scores for three scales of work stress (unsuitable job, competition-dismissal anxiety, and work overload: P < 0.001, P < 0.01, and P < 0.05, respectively). These findings suggest that computer work has no significant effect on perceived work stress in computer engineers; on the other hand, age and school career do have effects. PMID- 8404770 TI - Epidemiological and clinical features of Minamata disease. AB - Minamata disease is methyl mercury intoxication from fish contaminated by a chemical factory in Minamata city. Based on the results of our regional survey, cardinal clinical features of the disease were clarified by a multivariant analysis of all symptoms in inhabitants in the polluted area. The clinical features were found to be essentially the same as those of Hunter Russell syndrome; however, some additional symptoms were also found. Those symptoms are influenced by many factors, such as degree of exposure and duration of pollution. The disposition of each inhabitant also plays a role in clinical manifestation. This analysis contributes to a correct individual diagnosis and to the correct estimation of patients in polluted areas. Long-term studies also uncovered a few inhabitants who claimed to have begun to experience some neurological symptoms after pollution ceased. These symptoms were attributed mainly to aging. As many inhabitants with mild neurological complaints were not easily diagnosed, a questionable borderline group should be postulated for social settlement of Minamata disease. The characteristics of Minamata disease are discussed and compared to cases of methyl mercury poisoning in other parts of the world. PMID- 8404771 TI - Effect of nickel sulfide on induction of interleukin-1 and phagocytic activity. AB - Rat peritoneal macrophages were treated with crystalline nickel sulfide (NiS), washed, and then tested for interleukin-1 (IL-1) production and phagocytosis of IgG-opsonized sheep red blood cells (SRBC). Pretreatment of peritoneal macrophages with crystalline NiS did not affect their ability to produce IL-1. In contrast, pretreatment of peritoneal macrophages with crystalline NiS inhibited the phagocytic activity of the peritoneal macrophage. This inhibitory activity was due to an effect on the internalization step of the phagocytic process. The binding step of this process was not affected by exposure to crystalline NiS. The overall number of Ig-SRBC that interacted with each macrophage was not different in treated or untreated cultures. This interaction is mediated through Fc receptors on the macrophage membrane. This result suggests that crystalline NiS does not modulate the expression of Fc receptors by peritoneal macrophages. The results described in this study suggest that crystalline NiS has a potent immunomodulatory effect on macrophage phagocytic activity. PMID- 8404772 TI - Pulmonary function changes in children associated with fine particulate matter. AB - During winter months many neighborhoods in the Seattle metropolitan area are heavily affected by particulate matter from residential wood burning. A study was conducted to investigate the relationship between fine particulate matter and pulmonary function in young children. The subjects were 326 elementary school children, including 24 asthmatics, who lived in an area with high particulate concentrations predominantly from residential wood burning. FEV1 and FVC were measured before, during and after the 1988-1989 and 1989-1990 winter heating seasons. Fine particulate matter was assessed using a light-scattering instrument. Analysis of the relationship between light scattering and lung function indicated that an increase in particulate air pollution was associated with a decline in asthmatic children's pulmonary function. FEV1 and FVC in the asthmatic children dropped an average of 34 and 37 ml respectively for each 10( 4) m-1 increase in sigma sp. This sigma sp increase corresponds to an increase in PM2.5 of 20 micrograms/m3. It is concluded that fine particulate matter from wood burning is significantly associated with acute respiratory irritation in young asthmatic children. PMID- 8404773 TI - Environmental tobacco smoke exposure as determined by cotinine in black and white young adults: the CARDIA Study. AB - Biologic markers have provided a direct method for assessing exposure to environmental tobacco smoke (ETS) and yet few studies have used these techniques to document exposure in general samples of nonsmokers. Exposure to ETS was assessed by serum cotinine and self-report in 3300 nonsmoking participants in the CARDIA study. Nonsmoking status was validated by a cotinine level of < 14 ng/ml. Twenty-eight percent of the 18- to 30-year-olds were exposed to ETS as determined by a detectable serum cotinine level (2-13 ng/ml); prevalence of exposure was higher among blacks than whites (32% vs 24%, P < 0.001). Similarly, ETS exposure as defined by self-report (hours/week) was higher in blacks, particularly for exposure in the home and in other small areas. Multivariate predictors of cotinine-determined exposure included reported exposure, male gender, lower education, past smoking history, and spending time with smokers. Only among current users of marijuana, 20% of the sample, was the black race found to be an independent predictor of exposure. The prevalence of ETS exposure is higher in blacks than whites, as documented by self-report and confirmed by serum cotinine levels. Other correlates of exposure include demographic factors and factors which may be surrogate measures of exposure. PMID- 8404774 TI - An international perspective in neurobehavioral toxicology. PMID- 8404775 TI - Depressive states in workers using computers. AB - There have been few reports investigating the depressive states in workers using computers. We describe the depressive states observed in workers using computers and discuss the sources of their occupational stresses. The first subject is a 34 year-old male manager of a manufacturing company who had customarily worked until 9 PM. In 1985, it become necessary for him to work until midnight; symptoms of depression began to appear during this period, exacerbated after trouble with a computer. In 1986, he visited a psychiatrist and his condition was diagnosed as Major Depression according to DSM-III. The second subject is a 26-year-old male VDT (visual display terminal) operator in a general hospital. Before the onset, he had had to work until 8 PM and, at the end of each month, until midnight. Two months later, he became depressed and his condition was diagnosed as Major Depression according to DSM-III. The third subject is a 32-year-old male chief in the computer programming section of a bank. He had had to work until 8 PM, became depressed, and visited a psychiatrist who diagnosed his condition as Major Depression according to DSM-III. The authors discuss these cases from the standpoint of occupational stresses, as they are associated with work overload, and the important role these stresses played in the onset of the workers' depressive states. PMID- 8404776 TI - Effects of lead exposure on neurophysiological parameters. AB - To clarify the chronic effect of lead exposure on the central and peripheral nervous systems (CNS and PNS), we performed neurophysiological tests on 41 lead exposed male workers. Unexposed workers (controls, N = 39) were examined for auditory brain stem response (ABR), and their ABR parameters were compared with those of 15 lead-exposed workers age-matched to the controls. Neurophysiological tests included those of motor and sensory nerve conduction velocity of the radial nerve (MCV, SCVwa and SCVfw), electroretinograms, pattern reversal visual evoked potential (VEP), ABR, and short-latency somatosensory evoked potentials (SLSEP). Neurophysiological parameters were analyzed by regression analysis [independent parameters: age, exposure duration, and current and time-weighted average lead concentration in whole blood (PbB and TWA-PbB)]. ABR parameters were also tested by Student's t test. Significant negative correlations were found between radial MCV and TWA-PbB and SCVwa and PbB, while significant positive correlations were found between the latency of component N145 of VEP and exposure duration and between the latency of component N20 of SLSEP and PbB. The mean of interpeak latency between component III and V of ABR of 15 lead-exposed workers was significantly prolonged compared with that of the control group. These results suggested that lead exposure has a greater effect on the conduction function in the PNS than in that of the CNS in somatosensory and auditory pathways, and inversely in visual pathway. PMID- 8404777 TI - Acute neurobehavioral effects of co-inhalation of toluene and n-hexane on schedule-controlled behavior in rats. AB - Whether coexposure to toluene and n-hexane had any combined effects on the shock avoidance performance in rats was studied. Eighteen Wistar male rats with an avoidance rate of over 80% were selected and divided to three groups based on performance and body weight: (1) toluene, (2) n-hexane, and (3) toluene + n hexane. Each group was exposed alternately first to air and then to a particular organic solvent for 4 hr at various concentrations (50, 100, 200, 400, or 800 ppm, in ascending order). The effects of each organic solvent were evaluated by comparing the performance of rats during and after exposure with their own performance under the sham exposure to air by three-way ANOVA. The main results were that (1) 200, 400, or 800 ppm toluene exposures increased lever press rates, (2) 50 ppm n-hexane exposure decreased lever press and avoidance rates in a transitory manner and 800 ppm n-hexane exposure increased the lever press rate, (3) the 50 ppm mixture (25 ppm toluene + 25 ppm n-hexane) decreased lever press and avoidance rates persistently during and after the 4-hr exposure and the 800 ppm mixture (400 ppm toluene + 400 ppm n-hexane) decreased lever press and avoidance rates unpredictably when compared to the results of 400 or 800 ppm of toluene or n-hexane alone. In conclusion, n-hexane showed narcotic effects at 800 ppm and modified the acute neurobehavioral effects of toluene in rats at 400 ppm toward unpredictable results. PMID- 8404778 TI - Nerve-specific marker proteins as indicators of organic solvent neurotoxicity. AB - Effects of chronic exposure to n-hexane and toluene on some nerve-specific marker proteins in rat central nervous system (CNS) and peripheral nervous system (PNS) were assessed and compared. The rats were exposed to 2000 ppm n-hexane, 12 hr/day, 6 days/week, for 24 weeks, and to 1000 ppm toluene, 8 hr/day, 6 days/week, for 16 weeks. The level of neuron-specific enolase (NSE), creatine kinase-B (CK-B), and beta-S100 protein in cortex, cerebellum, spinal cord, and proximal and distal sciatic nerve was determined by enzyme immunoassay method. In n-hexane-exposed rats, the level of NSE, CK-B, and beta-S100 decreased significantly in the distal segment of the sciatic nerve, while the marker proteins in CNS and proximal sciatic nerve remained unchanged. In contrast, chronic exposure to toluene mostly affected these marker proteins in CNS tissues, displaying the increase of NSE, CK-B, beta-S100 in cerebellum, as well as the increase of beta-S100 in spinal cord. No quantitative changes of the three proteins in distal sciatic nerve were observed after exposure to toluene. n Hexane-induced peripheral distal neuropathy and toluene-induced brain gliosis appeared to be responsible for this different pattern of biochemical changes. The present study suggests the usefulness of using these nerve-specific marker proteins to assess the solvent-related CNS and PNS neurotoxicity. PMID- 8404779 TI - Workplace strategies for the control of work-related risks. AB - The prevention of work-related diseases and disorders, including those of neuropsychobehavioral nature, calls for integrated action to improve both job content and the working environment. Recent international labor standards have highlighted the importance of a good safety and health management system that ensures the assessment of risks and the appropriate control of these risks. Internationally, programs to cope with work-related risks place an increasing emphasis on participatory management of preventive measures, practical methods of risk assessment, and immediate improvements with the support of action-oriented training and information. Psychosocial factors, work performance, and job content need special attention. Workplace strategies must be developed by active participation of employers and workers, with their informed consent. These strategies and linked training activities should build on local practice and achievements rather than administrative models and be geared to the real local needs. In coping with the neurobehavioral effects, international sharing of positive experiences is useful especially with respect to: (i) providing information on potential neurobehavioral effects including labeling, safety datasheets, and precautions; (ii) providing practical advice about locally achieved improvements; (iii) providing action-oriented training; and (iv) creating opportunities for participatory workplace improvements. PMID- 8404780 TI - Review of air pollution and its health impact in Indonesia. AB - Air quality monitoring is part of the initial strategy in the pollution prevention program in Indonesia. Since 1978, the government of Indonesia has had a commitment to the World Health Organization (WHO) to provide air quality data for the Global Environmental Monitoring System (GEMS Programme)--The WHO/UNEP Project, in which certain cities from all over the world have been selected. Air quality as part of the WHO/UNEP project is monitored with respect to pollutants like SPM, SO2 and NOx. The result of the monitoring indicates that SPM and NOx are the predominant pollutants. Other pollutants such as O(x), H2S, NH3, and CO are also monitored in several big cities in Indonesia. The air pollution mainly comes from land transportation, industrial emissions, and a densely populated residential area where most people perform their activities. Review of the air pollution in Indonesia was based on the reports of the air quality monitoring in several large cities in Indonesia which covered air pollutants such as SPM, SO2, NOx, CO, O(x), and NH3 from 1978 until the latest available data in 1989. This review also discusses health impact investigations conducted in the community, especially from the exposure to SPM, CO, and lead from motor vehicle exhaust. PMID- 8404781 TI - A critical evaluation of the duplicate-portion technique in dietary surveys. PMID- 8404782 TI - Validity and reproducibility of a self-administered dietary questionnaire in obese and non-obese subjects. AB - The validity and reproducibility of a self-administered dietary questionnaire has been tested with specific attention to differences between obese and non-obese subjects. To test the validity, the dietary questionnaire was compared with 4-day food records, 24-h energy expenditure (24EE) and nitrogen excretion in 45 obese and 19 non-obese men and women. Energy intake was 2% higher (non-significantly) from questionnaire than from food records in the non-obese, but 35% higher (P < 0.001) in the obese. Comparing energy intake from the questionnaire with estimated 24EE, the questionnaire gave 4% higher values in both the non-obese and obese, differences which were not significant. The reproducibility in the obese sample that completed the questionnaire twice was comparable to that observed in normal populations. These data suggest that it is possible to obtain information on obese subjects' dietary intake that is at least as valid and reproducible as that from normal weight individuals. PMID- 8404783 TI - Alcohol consumption in relation to food intake and smoking habits in the Dutch National Food Consumption Survey. AB - The interrelationships between alcohol consumption, energy and food intake and smoking habits were studied in 1145 men and 1171 women, aged 22-49 years, in the Dutch National Food Consumption Survey, in which a 48-h dietary record method was used. The aim of the study was to investigate the effects of alcohol consumption on dietary habits and smoking. A strong relationship between alcohol consumption and energy intake was found. The energy derived from alcohol was not compensated for by lower intake of other nutrients. There was no increase in Quetelet's index with increasing alcohol consumption, except for non-smoking men who were heavy drinking on midweek days. Possible explanations for this apparent lack of an overall effect of alcohol calories are discussed. Alcohol consumption was much higher on weekend days than on midweek days. No differences in nutrient intake were found between non-drinkers, moderate drinkers and heavy drinkers on midweek days. On weekend days, however, there was a slightly higher total fat and saturated fat intake in moderately drinking men. For women cholesterol intake was found to be higher in moderate and heavy drinkers. Finally, a strong positive relationship between alcohol consumption and smoking was observed. It is concluded that the observations with respect to energy and nutrient intake and smoking habits are not indicative of a healthier lifestyle in moderate alcohol users between 22 and 49 years of age. Consequently, the more favourable prognosis of moderate drinkers cannot be ascribed to a more healthy lifestyle. PMID- 8404784 TI - Estimation of energy intake to feed subjects at energy balance as verified with doubly labelled water: a study in the elderly. AB - A study was intended to estimate energy intake (EI) for a nutrition intervention study. Subjects were 17 elderly men (age: 72 +/- 5 years) and 11 elderly women (age: 67 +/- 4 years). Two methods were used to measure EI: a 4-day dietary record (DR group; n = 12) or a dietary questionnaire (DQ group; n = 16). Subjects were fed for 3 weeks according to this intake during an intervention period, and energy expenditure (EE) was measured with doubly labelled water to verify the resulting figure for EI. Body weight and body composition were measured at the beginning of the second week and at the end of the third week, and metabolizable energy (ME) was calculated as gross energy intake (GEI) minus energy in faeces and urine. GEI, ME and EE did not differ between the DR and DQ groups (mean +/- SD for GEI: 10.09 +/- 1.21 and 9.29 +/- 1.36; ME: 9.09 +/- 1.28 and 8.34 +/- 1.31; EE: 10.13 +/- 1.57 and 9.25 +/- 0.35 MJ/d). In both groups ME was significantly lower than EE (DR group P < 0.05; DQ group P < 0.01). Body weight decreased significantly during the intervention period (mean +/- SD for DR group: -0.64 +/- 0.50 kg, P < 0.001; DQ group: -0.86 +/- 0.90 kg, P < 0.01). The change in body weight was significantly correlated with energy balance (ME - EE; P < 0.05). As shown from the results of the body composition measurements, body weight loss was a decrease of fat mass.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404785 TI - Comparative effects of prolonged intake of highly purified fish oils as ethyl ester or triglyceride on lipids, haemostasis and platelet function in normolipaemic men. AB - Conflicting results have been reported about the absorption of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) either as an ethyl ester (EE) or in a triglyceride (TG) formula. We decided to conduct a randomized double-blind study to compare the effects of EE and TG on plasma fatty acids, platelet function and haemostasis. Thirty-one healthy normolipaemic men were allocated to receive fish oil concentrate either as EE or TG with equal amounts of EPA (2.2 g and 2.2 g, respectively) and DHA (1.2 g and 1.4 g, respectively) or placebo daily for 7 weeks. Total cholesterol and the triglyceride level were not influenced differently by the two compounds. Repeated measurement ANOVA revealed a difference between TG and EE regarding incorporation of arachidonic acid (P = 0.034) and EPA (P = 0.007) into plasma cholesteryl esters. A discrimination not observed within plasma phospholipids. Both formulas had equal inhibitory effects on collagen-induced platelet aggregation and thromboxane B2 (TxB2) production in whole blood. Fibrinogen decreased 16% in EE (P = 0.034) and 12% in the placebo group (P = 0.11), but variance analysis of delta change during intervention did not indicate differences between groups. It is concluded that TG and EE fish oils are well incorporated into plasma lipids and have similarly beneficial influence one platelet function in men. PMID- 8404786 TI - Effects of pea and soybean fibre on postprandial lipaemia and lipoproteins in healthy adults. AB - To evaluate some possible mechanisms whereby total dietary fibre (TDF) may affect lipid metabolism in humans, six normolipidaemic males ingested on separate days a low-fibre test meal (2.8 g TDF) containing 70 g fat and 756 mg cholesterol, enriched with 10 g TDF in the form of either pea fibre or soybean fibre. Fasting and post-meal blood samples were obtained for 7 h and chylomicrons (CM) were isolated. Lipoproteins (VLDL+CM remnants, LDL, HDL) were isolated from the baseline samples and the samples of the 2-3 h triglyceride peaks. As compared to the postprandial response given by the control low-fibre test meal, adding fibre induced no change in serum glucose, insulin or Apo A1 and Apo B variations. The serum triglyceride response was not altered by adding fibres but the 2-3 h chylomicron triglyceride rise was increased (P < or = 0.05) by soybean fibre. VLDL+CM remnants, LDL and HDL triglyceride variations were unchanged with fibres. Cholesterolaemia decreased postprandially for 6 h, and was further lowered in the presence of pea fibre. This resulted from a marked decrease in serum esterified cholesterol. The chylomicron cholesterol and phospholipid rise was lowered in the presence of either fibre. The postprandial changes in the free cholesterol concentrations of the various lipoprotein classes were not altered by fibre whereas changes from baseline in esterified cholesterol concentrations were reduced by soybean fibre in LDL and amplified by soybean and pea fibres in HDL. The results obtained show that dietary fibre present in legumes may alter postprandial lipaemia and lipoproteins in humans to a variable extent. These effects could be related to some long-term metabolic effects. PMID- 8404787 TI - Non-starch polysaccharide/dietary fibre supplementation using small meals in long stay frail elderly patients. AB - Small meals of high non-starch polysaccharide (NSP)/dietary fibre cakes were offered to clinically stable continuing-care elderly patients, replacing the standard provision of low NSP/dietary fibre commercial cakes in an attempt to improve their well being. The study design was comprised of a 4 week control period, a 4 week intervention period, when modified cakes were offered, and finally a second 4 week control period. During intervention, an additional 33% NSP or 23% dietary fibre was offered, but only 7.7% and between 3.5% and 5.6% were consumed, respectively. Anthropometric measurements and bowel frequency did not change throughout the study but a decrease in aperient use was noted. From this study it was concluded that more comprehensive changes in food provision are required to achieve the current recommended consumption of NSP/dietary fibre in continuing-care elderly patients. PMID- 8404788 TI - Premature babies may achieve plasma ascorbic levels sufficiently high to inhibit antioxidant activity of plasma. PMID- 8404789 TI - Breakfast omission and cognitive performance of normal, wasted and stunted schoolchildren. AB - This study focuses on the effects of breakfast omission on cognitive performance. We studied 279 children from low socioeconomic level background ranging in age from 8 years 7 months to 10 years 11 months, categorized nutritionally as: normal, wasted or stunted. Evaluation comprised three cognitive tasks designed to be applied with a microcomputer. Assessment took place in their natural setting, after a mean of 14 h of overnight fasting, some having received a standard breakfast at random while the remaining children continued a fasting situation. We found no consistent association between study condition and performance in short-term visual memory, problem solving and attention tasks in any of the three nutritional groups. Stunted children showed significantly lower scores in the attention test irrespective of having received breakfast or not. These results suggest that given a motivating short-term task and maintaining routine conditions, missing breakfast does not affect the accuracy of the cognitive performance of children. Nutritionally affected children did not show a particular vulnerability to the fasting condition, but did show a specific cognitive deficit. PMID- 8404790 TI - Gastrointestinal effects of sugarbeet fiber and wheat bran in healthy men. AB - Gastrointestinal effects of feeding 20 g total dietary fiber as wheat bran (WB) and sugarbeet fiber (SBF) were compared in 17 healthy men in a randomized cross over design. The fibers were milled to similar particle sizes and consumed as supplements to the subjects' self-selected diets (SS). Transit times were similar among the diets. Fecal wet and dry weights were greater on the fiber diets compared to the SS diet (P < 0.05); wet weights were similar with WB and SBF; dry weights were greater with WB than SBF (P < 0.001); and moisture content was lower with WB than SBF (P < 0.01). Fecal pH was lower with WB than SBF (P < 0.025). WB and SBF provided similar fecal bulking effect; however, the contribution of fecal dry matter and fecal water content to increased bulk differed between fibers. PMID- 8404791 TI - The macronutrient composition of the Greek diet: estimates derived from six case control studies. AB - Nutritional data concerning the comparison (control) series of six case-control studies undertaken in the 1980s in Athens were used to estimate the percent energy intake from major macronutrient groups in the contemporary Greek diet. The data were generated by interviewer-administered semi-quantitative food frequency questionnaires in the context of studies probing the nutritional aetiology of several chronic diseases. The analyses were eventually based on 228 men and 610 women 40-79 years old. The main results were as follows: (i) age has very little effect on the proportion of energy intake from the macronutrients studied, although a declining trend is evident with respect to total energy intake; (ii) the proportion of energy intake from saturated fat and monounsaturated fat and, accordingly, total fat is higher among women than among men; (iii) in the Athenian (urban) Greek population in the 1980s, the proportion of energy derived from proteins is about 19%, from carbohydrates is about 37% and from fats is about 44%; and (iv) the P/S ratio is approximately 1:4 whereas the M/S ratio is approximately 1:1, although the latter ratio may be underestimated. It is concluded that low total fat intake is not a characteristic of the Greek diet and cannot explain the apparently health-promoting consequences of this diet. PMID- 8404792 TI - Converting Tanner-Whitehouse reference tricep and subscapular skinfold measurements to standard deviation scores. AB - The use of conventional standard deviation (SD) scores is inappropriate when data are not normally distributed, as is the case with the tricep and subscapular skinfolds. However, it is now possible to summarize centile data in three age specific parameters (called L, M and S) and by the appropriate use of these parameters, SD scores for skinfold measurements can be calculated that take into account skewness in the centile data. The L, M and S parameters for the tricep and subscapular skinfolds from age 1 to 19 years [J. M. Tanner & R. H. Whitehouse (1975) Arch. Dis. Childh. 50, 142-145] are presented and the equation for the calculation of SD scores described. PMID- 8404793 TI - Factors affecting iron status in non-pregnant women from urban south Finland. AB - Iron status was evaluated in 446 non-pregnant urban Finnish women, aged 17-50 years. The mean results (95% ranges) for blood haemoglobin concentrations, mean corpuscular volumes (MCV) and serum ferritin concentrations were 139 g/l (121 155), 89 fl (79-97) and 28 micrograms/l (7-120), respectively. Serum ferritin was < 12 and < 20 micrograms/l in 11% and 30% of the subjects, respectively. In analyses of covariance, high frequency of menstruation, prolonged menstrual bleeding time, blood donation and use of intrauterine devices had negative effects (P < 0.05) on iron status. Dietary habits or participation in very hard physical exercise did not affect iron status. Factors predicting low iron stores were evaluated in a logistic regression analysis. Blood donation (odds ratio and its 95% confidence interval: 2.53; 1.22-4.82) and very prolonged (> or = 8 days) menstrual bleeding (4.93; 1.63-14.9) increased the likelihood for low serum ferritin, whereas daily physical activity (0.02; 0.01-0.88) and use of multivitamin and mineral supplements (0.57; 0.33-0.98) were associated with higher concentration. In conclusion, blood losses increase the risk for iron depletion, while higher physical activity (presumably because of the positive effects on energy and iron intake) may prevent iron depletion. PMID- 8404794 TI - Resting metabolic rate and postprandial thermogenesis by level of aerobic fitness in young women. AB - Resting metabolic rate (RMR) and the thermic effect of a meal (TEM) were compared among women of three levels of aerobic fitness. Twenty-three euthyroid, eumenorrheic women (aged 18-35 years) were divided into three groups based on VO2 max standardized for fat-free weight (FFW), as determined from a graded exercise test: High Fit (HF): n = 7, VO2 max > 70 ml*kg FFW-1*min-1; Moderately Fit (MF): n = 8, VO2 max = 55-70 ml*kg FFW-1*min-1; and Low Fit (LF): n = 8, VO2 max < 55 ml*kg FFW-1*min-1). At 0700h RMR was measured for 1 h by indirect calorimetry with subjects in a fasted, preovulatory state, having refrained from exercise on the preceding day. The subject then consumed a liquid meal (12 kcal*kg FFW-1) and indirect calorimetry was continued for 3 h to determine the TEM. RMR adjusted for FFW using analysis of covariance was significantly higher (P < 0.05) in the HF group (mean +/- SEM = 1.08 +/- 0.03 kcal*min-1) compared to the MF (0.99 +/- 0.04) and LF (0.90 +/- 0.04) groups. Group differences in the thermic response did not reach statistical significance, although there was a trend for a high TEM in the HF group. There was a positive relationship between RMR and energy flux (average daily kcalorie intake + daily kcaloric expenditure in physical activity). These results suggest that women who exhibit high levels of exercise and aerobic fitness may be less energy efficient during the non-exercise portion of the day then their less active counterparts. PMID- 8404795 TI - Cultural and social acceptability of a healthy diet. AB - The extent to which the dietary practices recommended by nutrition science are compatible with an enjoyable lifestyle is a recurring theme in the debate on food and health in Denmark. The aim of this study was to see in practice what problems arise when ordinary people are confronted with a healthy diet. Fourteen of the participants in an 8 month dietary intervention study were interviewed about their opinions of, and experiences with, a diet composed in accordance with the Nordic nutrition recommendations. The interviews were qualitative, in depth and semistructured. The participants were interviewed twice, the first time towards the end of the intervention and again 3 months after the intervention ended. The interviews were transcribed verbatim and analysed thematically. For the participants, who were young students with a relatively high knowledge of nutrition, practical experience of a recommended diet was a series of surprises: the amount of food, its similarity to modern Danish food culture, its palatability, and the relatively small amount of dairy products in the diet were contrary to participants expectations. Participants found the recommended diet pleasant to live on, but expected certain economical and practical difficulties in applying it to everyday life outside the intervention. Hunger and satiety sensations changed and became more distinct. The results of the study indicate suggestions relevant for both industrial product development and nutrition information to the public. PMID- 8404796 TI - Prediction of twenty-four-hour energy expenditure in a respiration chamber in smokers and non-smokers. AB - Fourteen subjects (seven smokers, seven non-smokers) underwent at least one measurement of 24-h energy expenditure (EE) in a respiration chamber, smoking or not smoking as normal. Activity levels (AL) were calculated as multiples of basal metabolic rate (BMR) from records of time spent in specified activity categories and their average published energy costs. Predicted 24-h EE was estimated by multiplying AL by BMR measured on exit from the chamber. Smokers smoked an average of 18.6 cigarettes during the 24-h EE measurement (range 9-29). Measured 24-h EE was higher than predicted in the smokers (7.1%, P < 0.001) but not significantly different from predicted in the non-smokers (2.6% lower). We conclude that smoking increases 24-h EE and that factorial prediction of EE using average published energy costs of activities under-estimates 24-h EE in smokers but not in non-smokers. PMID- 8404797 TI - An evaluation of energy intakes in the 1983 Australian National Dietary Survey of Adults. PMID- 8404798 TI - Health psychology in the 21st century: acquired immunodeficiency syndrome as a harbinger of things to come. AB - An increasing focus on the human immunodeficiency virus (HIV) is important given the role of health psychology in preventing further spread of the epidemic and in maintaining quality of life in the estimated 1,500,000 Americans who are now infected. HIV presents health psychology with challenges reflecting 5 trends in medicine that have broad implications for the future, which extend beyond HIV: (a) the early identification of people who are at risk for disease, (b) the rising expectations for successful behavior change programs, (c) the growing populations of those who are coping with chronic disease, (d) the increasing shift to include community and public health perspectives, and (e) the emerging need to address health problems on a global scale. PMID- 8404799 TI - What underlies medical donor attitudes and behavior? AB - Donor attitudes, intentions, and behaviors have typically been conceptualized as organized along a bipolar continuum. This conceptualization is evident in I. G. Sarason et al.'s study of increasing participation in a bone-marrow registry in this issue. When the cumulative research on blood, bone-marrow, and organ donor behavior is considered, however, evidence suggests that a single, bipolar continuum may be insufficient and that a 2-dimensional (Positivity x Negativity) evaluative space may be minimally required to effectively represent and target the underlying substrates of donor behaviors. Negative beliefs and fears may constitute a particularly difficult obstacle to inducing donor behaviors and, thus, to promoting self-perceptions by people as donors. Understanding and changing these negative substrates, therefore, may be important if public health campaigns to increase donor behavior are to be cost-effective. PMID- 8404800 TI - Increasing participation of blood donors in a bone-marrow registry. AB - In an experiment to increase recruitment of unrelated bone-marrow donors, Ss were selected from a list of people who had donated blood within the past 24 months. They were randomly assigned to 3 groups. Members of the experimental group, 2 months before receiving a mailed brochure about a bone-marrow registry, were complimented on being blood donors and asked to complete a self-descriptive questionnaire. One control group received only the mailed brochure, and the other did not receive any mailing. The experimental group joined the registry at over 2 times the control-group rates. These results appear to be attributable to an attitude change associated with being recognized as a special group that contributed to the community's welfare. PMID- 8404801 TI - The influence of cancer patients' symptoms and functional states on patients' depression and family caregivers' reaction and depression. AB - In this article the paths among cancer patients' physical and mental health and the reactions and mental health of their family caregivers were examined. Data for these analyses came from a cross-sectional sample of cancer patients who were recruited through ambulatory outpatient chemotherapy units, and their family caregivers. Patients' depression was explained largely by their symptomatology and, to a lesser extent, by loss of mobility. Patients' physical limitations impacted caregivers' daily schedules but not their physical health. Patients' levels of depression were related to those of their caregivers. However, caregivers' optimism proved to be a significant predictor of their mental health and reactions to caregiving. PMID- 8404802 TI - Effects of aerobic exercise on cognitive and psychosocial functioning in patients with mild hypertension. AB - The effects of 16 weeks of physical exercise training on the psychological functioning of 90 patients with mild hypertension were examined. At baseline and after 16 weeks of training, patients completed a psychometric test battery that included objective measures of neuropsychological performance and standardized self-report measures of psychosocial functioning. Patients were randomly assigned to one of three groups: aerobic exercise, strength training and flexibility exercise, or a waiting list control group. After training, there were no group differences on any of the psychological measures, even though patients who engaged in exercise perceived themselves as functioning better in a number of psychological domains. PMID- 8404803 TI - Effects of differing intensities and formats of 12 months of exercise training on psychological outcomes in older adults. AB - The 12-month effects of exercise training on psychological outcomes in adults ages 50-65 years were evaluated. Ss (N = 357) were randomly assigned to assessment-only control or to higher intensity group, higher intensity home, or lower intensity home exercise training. Exercisers showed reductions in perceived stress and anxiety in relation to controls (p < .04). Reductions in stress were particularly notable in smokers. Regardless of program assignment, greater exercise participation was significantly related to less anxiety and fewer depressive symptoms, independent of changes in fitness or body weight (p < .05). It was concluded that neither a group format nor vigorous activity was essential in attaining psychological benefits from exercise training in healthy adults. PMID- 8404804 TI - Self- and spouse ratings of anger and hostility as predictors of coronary heart disease. AB - Self- and spouse ratings of anger and hostility were examined as predictors of coronary heart disease (CHD) in 185 cardiac patients. Patients completed the Multidimensional Anger Inventory (MAI) and the Marlowe-Crowne Social Desirability (MCSD) Scale; the MAI (rewritten to 3rd person) was completed by patients' spouses or by a peer. Thallium scans were used to measure CHD status. Results showed that patient-rated MAI scores were inversely correlated with MCSD. There were no gender differences for patient-rated MAI scores, but spouse ratings showed gender effects for Anger-Arousal and Hostile Outlook: Women rated their husband higher than men rated their wife. Patients with positive thallium scans were no different from those without CHD on patient-rated MAI scores; however, spouse ratings indicated that those with CHD had higher Hostile Outlook and Anger In scores. After accounting for the effects of traditional CHD risk factors, only spouse-rated hostility contributed significant incremental variance to the prediction of CHD status. PMID- 8404805 TI - Effects of prebehavioral cognitive work on adolescents' acceptance of condoms. AB - Two educational strategies designed to promote condom use for sexually transmitted disease protection were tested in a field experiment involving 291 female, adolescent family planning clinic clients. The 1st strategy was designed to enhance attitude-behavior correspondence by increasing direct experience with handling condoms. The 2nd, a contingency-planning exercise, induced clients to generate a mental representation of negotiating condom use with a sexual partner. Both strategies were compared with the standard education. The dependent measures were condom acceptance (operationalized by the number of condoms taken), attitudes, and knowledge. Clients in the contingency-planning condition accepted about 60% more condoms than did other clients. Condom attitudes followed the same pattern, and knowledge did not differ among conditions. PMID- 8404806 TI - Perceptions of risk for AIDS among women in drug treatment. AB - In this article factors associated with the self-perceived risk for acquired immunodeficiency syndrome (AIDS) were examined using data obtained from a cross sectional survey of 155 current and former drug-using women in methadone maintenance. Results suggest that drug-involved women are realistic in their self perceptions of AIDS risk with respect to intravenous (IV) drug-using behavior but underestimate their risk from sexual activity. Perceived risk was associated with current IV drug use, duration of sexual relationship, and partner's nonsupportiveness. Partner's serostatus or history of IV drug use was unrelated to risk perception, as were multiple partners, anal sex, prostitution, and the nonuse of condoms. Implications of these findings for designing interventions for drug-involved women are considered. PMID- 8404807 TI - Testing four competing theories of health-protective behavior. AB - Four competing theories of health-protective behavior are reviewed: the health belief model, the theory of reasoned action, protection motivation theory, and subjective expected utility theory. In spite of their commonalities, these models are seldom tested against one another. The review points out the similarities and differences among these theories and the data and analyses needed to compare them. In addition to describing the content of the models, their conceptualization of key variables, and the combinatorial rules used to make predictions, some general problems in theory development and testing for health behaviors are examined. The article's goal is to help investigators design studies that will clarify the strengths and weaknesses of these models, leading toward a better understanding of health behavior. PMID- 8404808 TI - Theoretical studies on the mobility-shift behavior of binary protein-DNA complexes. AB - The theory of mass transport coupled to reversible interactions under chemical kinetic control forms the basis for computer simulation of the electrophoretic mobility-shift behavior of binary protein-DNA complexes. Several systems have been modeled in terms of either (i) specific binding of a protein molecule to a single site on the DNA molecule; (ii) cooperative binding to two or three sites; (iii) noncooperative binding to two sites, both of which bind protein with equal affinity; (iv) statistical binding to multiple sites having identical intrinsic binding constants; or (v) protein-induced DNA loop formation. Both models (iii) and (v) embody the concept of reversible isomerization of protein-DNA complexes. The resulting simulations have provided fundamental information concerning (i) the factors governing the electrophoretic persistence and separation of protein DNA complexes; (ii) the shape of experimental mobility-shift patterns; (iii) the generation of the protein-DNA ladder upon titration, for example, of the 203-base pair operator with lac repressor; and (iv) the theoretical bases for quantitative interpretation of the patterns in terms of thermodynamic and kinetic parameters. The practical implications of these findings are discussed. PMID- 8404809 TI - A new concept for separating nucleic acids by electrophoresis in solution using hybrid synthetic end labelled-nucleic acid molecules. AB - Analogy between the symmetry breaking of the electrical driving force and the opposing friction force in gels using pulsed electric fields is made with the corresponding effect for polyelectrolyte coils in solution related to the molecular weight-independent charge density. The synthesis of hybrid molecules to break the symmetry of constant charge density is proposed, in which standard polypeptide end labels are attached to nucleic acid fragments. The combinatorial chemical library scheme of Brenner and Lerner (Proc. Natl. Acad. Sci. USA 1992, 89, 5381-5383), making use of hybrid molecules for another purpose, is mentioned, with the suggestion that for electrophoresis in solution a neutral end-labelled polypeptide chain would give the largest molecular weight and time-dependent change in the charge density for pulsed fields. PMID- 8404810 TI - Detection of DNA damage-recognition proteins using the band-shift assay and southwestern hybridization. AB - We describe electrophoresis and biochemical conditions that allow detection of damaged DNA-binding proteins in cell extracts. In addition, we present an overview of the damage-recognition DNA-binding proteins from eukaryotic cells and discuss their hypothetical role in DNA repair. PMID- 8404811 TI - Electrophoretic analysis of protein-induced DNA bending and twist changes. AB - DNA fragments with a varied phasing of two intrinsic bends at their ends and a tet operator in-between were constructed to determine Tet repressor-induced twist changes in the operator DNA. These distance variants show a sinusoidal dependence of their electrophoretic mobilities on the phasing of their bends in polyacrylamide gels. Complex formation with Tet repressor leads to a displacement of the first minimum, indicating an unwinding of the tet operator DNA. Model calculations were performed to assess the contribution of Tet repressor-induced DNA bending to this result. They revealed that the amplitude of the electrophoretic mobilities of the distance variants may be used as a parameter to separate the effects of Tet repressor-induced twist change and bending. The systematic analysis presented here may help to improve the quantitative interpretation of gel shifts and promote the use of this highly sensitive method to gain structural information about protein-DNA complexes. PMID- 8404812 TI - A simple physical model for the gel electrophoretic manifestations of sequence dependent DNA superstructures. AB - A simple physical model of the gel electrophoretic manifestations of sequence dependent DNA superstructures is illustrated. The model is based on the calculation of the curvature dispersion as evaluated by integrating the orientational parameters of the base pairs along the chain. Such a quantity is proportional to the straightening activation energy of the curved DNA in the direction of the average orientation of the helical axis and was found to be proportional to the logarithm of the gel electrophoretic retardation. The model is capable of consistently explaining the different electrophoretic manifestations of DNA superstructures, such as the retardation of a large ensemble of multimeric oligonucleotides with different sequences, periodicities, and lengths, and the permutation gel assays as well as the mobility changes consequent to extensive point mutations in a DNA tract. PMID- 8404813 TI - Effects of anomalous migration and DNA to protein ratios on resolution of equilibrium constants from gel mobility-shift assays. AB - Numerical resolution of binding constants from data generated by the gel mobility shift assay is dependent on the experimental resolution of the different ligation states of the DNA. Previously we showed that the populations of the intermediate ligation states at partial saturation with the protein ligand are extremely sensitive to cooperativity (Senear, D. F. and Brenowitz, M. J. Biol. Chem. 1991, 266, 13661-13671). This makes accurate gel mobility-shift data extremely useful to the demonstration of cooperativity. However, the accuracy with which the intermediate ligation state populations are resolved has been questioned. Thus, two additional and related questions are now considered. First, what information is available if the intermediate ligation state populations are not used in the analysis of binding constants. Second, is accurate information obtained from those states under conditions of high DNA concentration. These questions are addressed by using the interactions of the lambda cI repressor protein with the three site operator, OR, and the interaction of the E. coli GalR protein with the single site operator, OE. Both simulated and experimental data are analyzed. The results point to two conclusions. First, precise resolution of all macroscopic constants for binding of proteins to DNA is critically dependent on the intermediate ligation state populations; resolution is limited to at most two DNA sites if these states are not used in the analysis. Second, when the DNA and protein concentrations used in the titrations are comparable, the resolution of binding constants is extremely sensitive to experimental uncertainty in the macromolecule concentrations. PMID- 8404814 TI - Analysis of trp repressor-DNA interactions using gel electrophoresis. AB - Quantitative analysis of the DNA-binding equilibria of E. coli trp repressor by gel electrophoresis led to reevaluation of our understanding of this complex system. In this review, the data leading to controversy about the trp system are discussed, and our current understanding is presented. PMID- 8404815 TI - Characterization of the electrophoretic properties of nucleosome core particles by transverse polyacrylamide pore gradient gel electrophoresis. AB - Transverse pore gradient gel electrophoresis, previously applied to bent DNA, has extended the usefulness of the gel retardation assay in two ways: (i) by differentiating between different DNA conformations; (ii) by providing information regarding the physical properties of DNA. In the present study, similarly extended information is obtained with regard to a well-characterized DNA-protein complex, the chicken erythrocyte nucleosome core particle. (i) The winding of DNA around the protein core constrains the DNA which renders its Ferguson curve (migration distance vs. gel concentration) similar to that of kinetoplast DNA, i.e. it intersects sharply with the Ferguson curves of linear DNA standards. By contrast, the deproteinized nucleosome DNA exhibits a Ferguson curve similar to linear standards of the same length. (ii) Interpretation of the Ferguson curve based on a mathematical model shows that the nucleosome exhibits a linear Ferguson plot [log(mobility) vs. gel concentration]. This is similar to and characteristic of spherical proteins, contrasting with the concave plot typical for linear and bent DNA. (iii) The effective size of the nucleosome, evaluated in terms of an "equivalent sphere" (i.e. a hypothetical spherical particle with a radius, Res, having the same electrophoretic mobility as DNA for a particular set of experimental conditions), remains invariant across the gel concentration range of 3-9%T. This is similar to proteins and bacteriophages and contrasts with the progressive decline of Res with increasing gel concentration observed for linear DNA and the deproteinized nucleosomal DNA. PMID- 8404816 TI - Measurement of binding kinetics using the gel electrophoresis mobility shift assay. AB - The gel electrophoresis mobility shift assay is a technique for the qualitative and quantitative analysis of protein-DNA complexes. The ability to resolve reactants, reaction intermediates and products makes this method particularly well-suited for the measurement of the assembly and dissociation rates of protein nucleic acid complexes. Here we identify conditions that must be met and variations of the technique that are useful for the measurement of reaction rates. PMID- 8404817 TI - Studies of DNA bending and flexibility using gel electrophoresis. AB - Gel electrophoretic methods have become established as primary tools in the study and elucidation of sequence-directed curvature both in free DNA and in the operator DNA of several site-specific nucleoprotein complexes. Results using them have been generally consistent with physical methods sensitive to DNA structure and conformation in those instances where direct comparisons can be made, and in a number of cases, gel methods have provided unique information not presently available from other techniques. Two basic strategies have been used: one based upon anomalous gel mobility effects; and a second based upon cyclization properties of curved DNA. Within each of these categories, various approaches have been used, some of which can lead, in favorable cases, to quantitative estimation of bending angles. In this review, the various gel-based methods that have been used to date are critically discussed and the qualitative and quantitative information that can be obtained from them is evaluated. A number of possible structural models for DNA curvature are described and a distinction is drawn between static or fixed bending and bending due to anisotropic flexibility at specific sequence loci. The importance and roles of gel electrophoretic methods in providing experimental approaches to this question are discussed. It is suggested that both static curvature and anisotropic flexibility in operator DNA may provide much of the basis for indirect readout of sequence information by specific site-binding regulatory proteins. PMID- 8404818 TI - Identification of predominant T-cell receptor rearrangements by temperature gradient gel electrophoresis and automated DNA sequencing. AB - To assess the diversity of T-cell receptor (TCR) gene rearrangements in uncloned lymphocytes we used a three-stage strategy that allows the detection of a restricted TCR repertoire and the identification of the predominant, rearranged sequence(s). We have analyzed in parallel T cells obtained from a renal cell carcinoma infiltrate that specifically lyse the autologous tumor after in vitro culture and T cells from autologous peripheral blood. First, DNA amplification by the polymerase chain reaction (PCR) was performed with a number of oligonucleotide primers specific for several TCR V alpha gene families. All V alpha primers displayed specific amplification products in the peripheral blood, while a restricted TCR repertoire was present in the tumor-infiltrating lymphocytes. Subsequently, positively amplified PCR products were run in a temperature-gradient gel electrophoresis. A limited number of bands corresponding to predominant homo- and heteroduplexes was only found in the tumor-infiltrating lymphocytes. The presence of a low number of rearranged TCR sequences in these samples was confirmed by automated single-stranded DNA sequencing using a single fluorescent dye. These results support the broad application of this strategy for targeted sequencing of those PCR products carrying predominant DNA templates without previous DNA cloning. PMID- 8404819 TI - Analysis of plasmid-DNA and cell protein of recombinant Escherichia coli using capillary gel electrophoresis. AB - Plasmid DNA prepared from cultivation samples of recombinant Escherichia coli was analyzed by capillary gel electrophoresis. We used this method to control the genetic stability during a fed-batch culture. The plasmid DNA and a standard DNA mixture with molecular weights in the size range of 3000-22,000 base pairs (bp) were analyzed in capillaries that were filled with solutions of non-cross-linked polyacrylamide. With this method the plasmid DNA from recombinant E. coli cultivation samples was analyzed within 30 min. Separation parameters such as gel concentration, capillary length and current were optimized for that purpose. The method was modified to allow the analysis of proteins. Crude cell lysate samples could be screened for the protein pattern within 15 min in sodium dodecyl sulfate polyacrylamide filled capillaries. The method was also used to verify product purity after the down-stream process. PMID- 8404820 TI - A traveling-wave micropump for aqueous solutions: comparison of 1 g and microgram results. AB - The operation of a micro-chip-based fluid circulator based on the interaction of a high-frequency traveling wave with a thermally generated inhomogeneity in an aqueous medium is investigated. The profiles of the electric field, the thermal gradient and the driving force within the device are derived numerically, and the dependence upon applied voltage under different experimental conditions is derived. The importance of convection in the operation of the device is assessed by operating it in various orientations, as well as under microgravity, and also by investigating the reversal and turn-on characteristics. The pump showed the unexpected ability to trap microparticles present in the driven fluid. PMID- 8404821 TI - Lane distortions in gel electrophoresis patterns. AB - Distortions effects were studied as a function of gel environment, apparatus design and buffer type. Outward lanes distortions pronounced in low conductivity buffers for both continuous buffer systems and stacking gels, were field strength dependent. In discontinuous buffer systems, the moving boundary of the Laemmli buffer system deformed depending on the environment. In the Michov buffer system, the high conductivity resolving gel surrounded by a low conductivity electrode buffer, displayed straight pathways until a field strength of 15 V/cm, permitting to obtain information about lanes distortions depending on the acrylamide matrix structure. A decreased ammonium persulfate concentration, e.g. to 0.03%, induced phase segregation in gels at low temperature during the run: a critical endpoint near 11 degrees C for a gel in the Michov buffer system under conditions of electrophoresis was supposed. Gel collapse resulted in inward deviations in the lanes of the pattern and also faint bands in the zones where the gel was retracted. Besides this effect of catalyst concentration, the inwardly distorted pattern depended on temperature, field strength, solvent and hydrolysis due to gel aging. Amendments for overcoming lane distortions are suggested. PMID- 8404822 TI - Temperature-gradient gel electrophoresis for analysis and screening of thermostable proteases. AB - The thermal unfolding of microbial serine proteases was studied by temperature gradient gel electrophoresis (TGGE). Conditions for a native polyacrylamide gel electrophoresis were established, and the temperature gradient was applied perpendicularly to the direction of electrophoretic migration. Mobility changes of the protease molecules were indicative for thermally induced conformational changes. The transition temperature was determined with good accuracy. The native and active protease conformation was detected by an activity assay in the temperature-gradient gel. As a consequence of the typical protease autoproteolytic reaction at elevated temperatures, the unfolded protease conformation could not be detected for non-inhibited, active subtilisin. After inhibition by phenylmethylsulfonyl fluoride (PMSF) the complete structural transition could be followed by TGGE. This transition is "discontinuous", i.e. the thermal transition is either very slow, compared to the time of electrophoresis, or irreversible, as known for subtilisins from calorimetric data. Inhibition by the strong serine specific inhibitor diisopropyl fluorophosphate (DFP) led to two conformations at low temperature. One conformation is stabilized by 8 degrees C, the other by at least 20 degrees C as compared with PMSF inhibition. The influence of calcium ions on the subtilisin stability was investigated by a series of TGGE under different calcium concentrations. The strong calcium binding site is occupied even without added calcium, occupation of the weak binding site leads to a stabilization of 10 degrees C with a binding constant around 10(6) M-1. The subtilisin Carlsberg stability could also be investigated in unpurified bacterial culture supernatants. Thus, the method is suitable for screening of thermostable subtilisin mutants directly after expression in a bacterial host.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404823 TI - Electrotransfer of fixed phosphoproteins from pieces of dried polyacrylamide gel to small disks of nitrocellulose, nylon or polyvinylidene difluoride. AB - A simple method for the transfer of 32P-labeled proteins from dried polyacrylamide gels to small disks of nitrocellulose, nylon or polyvinylidene difluoride (PVDF) is described. Gel pieces containing the desired phosphoprotein are rehydrated in buffer containing sodium dodecyl sulfate (SDS) and sealed in agarose in a glass tube over a supporting gel of polyacrylamide. Protein is transferred upwards through a discontinuous density gradient of SDS-buffer and methanol to a disk of membrane sealed to the mouth of the tube with dialysis membrane. The method allows the concentration of a phosphoprotein present in several gel pieces to a single disk of immobilized membrane. Recovery of phosphoprotein was at least as good as obtained with conventional electroblotting. Application of the method to the analysis of the phosphoamino acid content of the astrocyte marker, glial fibrillary acidic protein, is described. PMID- 8404824 TI - Horizontal semi-dry electroblotting for the detection of the low density lipoprotein receptor in solubilized liver membranes. AB - A high efficiency transfer of the low density lipoprotein (LDL) receptor proteins from polyacrylamide slab gel onto immobilizing nitrocellulose membranes using the horizontal semi-dry electrophoretic system is described. The transfer of the LDL receptors from solubilized rat liver microsomes was performed between two graphite plate electrodes in a continuous buffer system containing methanol and sodium dodecyl sulfate. The protein transfer was achieved in only 150 min at a constant current of 0.8 mA/cm2 at room temperature with very low Joule heat development. The homogeneous electric field yield between the two electrode plates produced a satisfactory transfer of the LDL-receptor protein band in spite of its high molecular weight, and only few protein traces remained in the polyacrylamide gel after blotting. This improved method allows a rapid and quantitative transfer of the LDL receptors without protein denaturation, since the specific binding activity of the blotted receptor is retained as demonstrated by ligand-blotting and immunoblotting. PMID- 8404825 TI - Electrophoretic characterization of wheat grain allergens from different cultivars involved in bakers' asthma. AB - In order to identify and characterize the wheat grain allergens involved in bakers' asthma, proteins were sequentially extracted from whole-meal flour. The polypeptide composition of the individual solubility fractions (albumin/globulin, gliadin and glutenin) was analyzed by sodium dodecyl sulfate-gel electrophoresis (SDS-PAGE), and high-resolution two-dimensional gel electrophoresis with immobilized pH gradient 4-9 in the first dimension (IPG-Dalt). The resolved polypeptides were transferred onto an immobilizing polyvinylidene difluoride membrane and incubated with a pooled serum from four asthmatic bakers. Bound IgE was demonstrated by autoradiography using 125I-labeled anti-human IgE. Our study demonstrated that the serum of the bakers allergic to flour contained IgE antibodies which bound to numerous polypeptides of all three solubility fractions. The highest percentage of IgE binding was observed with certain albumin and/or globulin polypeptides, whereas the gliadins and glutenins exhibited considerably less allergenicity. SDS-PAGE revealed that the protein which bound the highest percentage of IgE from the sera of the allergic bakers is a 27 kDa albumin. More detailed investigations using IPG-Dalt demonstrated that this allergen is not a single polypeptide but consists of several polypeptide spots that differ in their isoelectric points. Quantitative studies using computer-assisted laser densitometry revealed that the amount of patients' IgE bound by these particular polypeptides differed considerably between the seven wheat cultivars examined, ranging from 13% to 53% of the total radioactive uptake. PMID- 8404826 TI - Purification of phosphoenolpyruvate carboxylase subunits and isoforms from Vicia faba L. by preparative gel electrophoresis and their detection by enzyme-linked immunosorbent assay. AB - A purification procedure which yields nearly homogenous subunits of stomatal phosphoenolpyruvate carboxylase from epidermal strips of Vicia faba L. is reported. Preparative gel electrophoresis was found to be the most suitable technique for subunit purification. Denatured subunits of the stomatal enzyme in the eluate were immunologically detected by enzyme-linked immunosorbent assay (ELISA) tests. The enzyme preparation meets all requirements for its use in antibody production. PMID- 8404827 TI - Differentiation of proteins in polyacrylamide gels by a modification of silver staining for the PhastSystem and a laser densitometer. AB - Nonspecific background staining of the gel matrix is the limiting factor in the differentiation of proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. It is caused by nonspecific binding of silver ions in the gel matrix, either to nonprotein compounds or through the chemistry of the polyacrylamide gel itself. The resulting stain of the gel produces a stain baseline, making differentiation of protein bands with a laser densitometer difficult. We have therefore developed a modified silver staining method for the PhastSystem to reduce such nonspecific background staining. Apart from slowing the development program by lowering the temperature to 15 degrees C and shortening the incubation time to 4 min, the essential step in the modification is the combined use of an EDTA and Tris-acetate buffer solution which stops the reduction of silver ions. The reduced background staining leads to an improved detection of protein bands and virtually identical zero lines for the laser densitograms and the stain baselines. The total staining time is 91 min. All steps in the program are fully automated and continuous, employing the PhastSystem staining unit. PMID- 8404828 TI - Capillary electrophoresis quantitation of l-L-folinic acid in the presence of its inactive d-L-form. AB - The diastereomers d-L and l-L-folinic acid were separated by capillary electrophoresis, using heptakis (2,3-di-O-methyl)-beta-cyclodextrin as a chiral active component in the background electrolyte. The method proved suitable for the quantitation of these compounds in one commercial pharmaceutical preparation. PMID- 8404829 TI - Simultaneous determination of native and subunit molecular weights of proteins by pore limit electrophoresis and restricted use of sodium dodecyl sulfate. AB - In the present communication, we describe a method for the determination of both the native and subunit molecular weight of a protein by a single pore limit electrophoretic run. When native proteins and sodium dodecyl sulfate (SDS)-heat denatured proteins were electrophoresed in a 4-28% gradient-polyacrylamide gel, in the presence of 0.02% SDS in running buffer, their respective molecular weights (native and subunit) could be determined by comparing the migration distances of unknown and standard proteins. The presence of SDS up to 0.02% in the running buffer did not dissociate the native proteins into their subunits. The linearity in distribution of native and SDS-polyacrylamide gel electrophoresis (PAGE) standard marker proteins used for the molecular weight determination further showed the efficacy of the present technique. PMID- 8404830 TI - Sleepiness in long distance truck driving: an ambulatory EEG study of night driving. AB - Eighteen truck drivers had their EEG recorded continuously during a night or evening drive between southern Sweden and Stockholm (500 km). They also carried out self ratings of sleepiness and performance every hour. The EEG was subjected to spectral analysis. The drivers were divided into two groups with a night group (n = 7) who drove between 20:30 to 97:20 and an evening group (n = 11) who drove between 18:20 and 04:00. The night group showed higher subjective sleepiness and lower subjective performance, and increased alpha and theta burst activity during the last three hours of the drive. The groups did not differ for the first 2-3 h of the drive. For the night group, a significant intraindividual correlation was found between subjective sleepiness and EEG alpha burst activity. End-of-the drive subjective sleepiness and alpha burst activity were significantly correlated with total work hours and arrival time but not with age, diurnal type, prior (rated) sleep length, total break time, drive time or prior time awake. A regression analysis showed that total work hours and total break time predicted 66% of the variance of alpha burst activity during the end of the drive. PMID- 8404831 TI - Human vigilance in railway and long-haul flight operation. AB - Human operators in transport operations are often confronted with monotony, boredom, and irregular work schedules. This situation has become increasingly more acute because of the growing automation of systems. This paper presents methodology and preliminary results for two field studies on the vigilance of train drivers and long-range aircrews. The aim of these studies was to identify factors that can modify vigilance and to elaborate several specific solutions for reactivation. The method is based on the collection of physiological data in the field and on task observation of the operators. The recorded physiological data (EEG, EOG, EKG) permit an evaluation of vigilance and mental workload. The rest activity cycles are estimated by actometry. The use of EEG and EOG are discussed in relation to monotony and sleep deprivation. For pilots, results show a high occurrence of decreased vigilance, particularly during phases of low workload (i.e., when cruising). Furthermore, it was shown that these periods of lowered vigilance can occur at the same time for two crew members. A great number of incidents of decreased vigilance were also observed for the train drivers. These incidents occurred even though the operators sometimes had high levels of activity. A direct relation was also noted between sleep duration and the onset of rest. These studies provide several means for maintaining vigilance during activities and improving the system of work schedule rotation. PMID- 8404832 TI - A psychophysiological approach to the assessment of work underload. AB - The findings of a laboratory investigation of the relationship between the subjective and physiological components of work underload are reported. The subjective component is described in terms of the subjective work underload checklist, mental effort, and cognitive arousal. The physiological component is defined in terms of heart rate and heart rate variability. Evidence for an increase in work underload with a decrease in heart rate is provided. The relevance of this research to the aerospace environment is discussed and the need to investigate the behavioural component of work underload emphasized. PMID- 8404833 TI - Heart rate and workload variations in actual and simulated flight. AB - The monitoring of heart rate in aviation research provides a global index of pilot workload. Heart rate variability is a promising measure but more complex to assess and therefore less often used, especially in dynamic task environments. The application of spectral analysis techniques of heart rate for workload assessment in aviation is illustrated and discussed. Ideally, workload assessment includes real-time or continuous measures to reveal variations in workload as pilots accomplish their tasks. Four studies are discussed that used heart rate measures for the assessment of: (1) training effectiveness; (2) stress of flight instruction; (3) simulator realism; and (4) flight task difficulty. Results indicate that cardiovascular measures are well suited to index different mental states of pilots as well as their dynamic responses to variations in workload. PMID- 8404834 TI - Heart rate as a psychophysiological measure for in-flight workload assessment. AB - The need to assess pilot workload during flight has become of increasing importance over the past decade--both in combat aircraft where workload can be excessive, and in civil transport aircraft where underload may occur. Subjective reporting in some form has been the long and well established method for assessing workload in the 'real world'. but because subjective opinions are vulnerable to bias and to preconceived notions, an additional measure can be of considerable value on occasions. Of the available psychophysiological variables, recording pilots' heart rates appears to be the most useful. This paper discusses the choice of this measure and presents examples from several studies in which workload was assessed in flight. PMID- 8404835 TI - The evaluation of the electrooculogram as a psychophysiological measuring instrument in the driver study of driver behaviour. AB - Certain disadvantages of the EOG (electrooculogram) in the measurement of gaze behaviour (for example lack of stability) can be overcome by using on-line computer identification of saccades and additional keyboard marking of relevant gazes by the experimenter. This reveals the advantages of EOG measurement for precise fixation durations of gazes on defined instruments, activation decrements and increments of drivers due to time on task or mental effort (via saccadic velocity), and blink behaviour as controlled interruptions of visual behaviour. Three studies of traffic relevant questions were performed: (a) to better define optimal characteristics of digital displays; (b) to evaluate possible misperceptions caused by reflexions in the outside rear view mirror images passing through double-gazed windows of the car; (c) and to compare a head-up display vs. conventional speedometer reading in real traffic situations. Saccades and blinks of the drivers in these situations were measured by the EOG-ON-LINE device, and its effectiveness assessed thus. PMID- 8404836 TI - Air-to-ground training missions: a psychophysiological workload analysis. AB - Psychophysiological measures are used to assess the workload of F4 Phantom aircraft pilots and weapon systems officers (WSOs) during air-to-ground training missions and during the performance of two levels of difficulty of a laboratory tracking task. The bombing range portion of the missions was associated with the highest pilot workload, while the WSO flying the aircraft was the highest workload segment for the WSOs. The pilots' data were found to have a wider range of values for the physiological measures than were found in the WSO data. The different levels of tracking task difficulty produced significant physiological effects but the range of values found for most of the flight segments were much greater. These data demonstrate that extrapolating laboratory data to the flight environment is risky at best. The various physiological measures were differentially sensitive to the different demands of the various flight segments. PMID- 8404837 TI - Biochemical and haemodynamic indicators of stress in truck drivers. AB - In order to investigate the extent of stress reaction during driving, this study was carried out on truck drivers engaged in long distance work. For each driver, epinephrine, norepinephrine, and cortisol were measured in urine, and dynamic ECG and blood pressure were monitored. The excretion rates of catecholamines showed great individual differences in the size of variations related to driving conditions. Epinephrine excretion rates were particularly high when weather and traffic conditions were more stressful. A relationship was found between epinephrine urinary levels and state-anxiety scores. Urinary excretion of norepinephrine was generally increased at the end of the working day and while driving in fog. The pattern of cortisol was not affected by the stress related to driving. The highest mean heart rates were monitored during difficult traffic and bad weather conditions. PMID- 8404838 TI - The use of psychophysiology to assess driver status. AB - Twenty subjects completed an on-the-road driving experiment, consisting of two different tests conducted on two separate days. A two-part test was administered while subjects were under the influence of alcohol (BAC < = 0.05%); a four-part test was administered without alcohol consisting of a 2.5 h driving test under vigilance conditions on a quiet highway. The order of the tests was balanced across subjects. Changes in relevant physiological parameters, such as ECG and EEG, reflected changes in driver status and predicted driving impairment. Impairment of driving performance was measured in a standard driving test (SD lateral position and SD steering wheel movements) and in a recently developed car following test (reaction to speed changes of a leading car). PMID- 8404839 TI - Evaluation of workload in air traffic controllers. AB - This study examined 20 air traffic controllers from the Rome Regional Air Control Centre for three successive work shifts: afternoon (13:00-20:00), morning (07:00 13:00) and night (20:00-07:00). The number of aircraft under control per hour was recorded as index of workload. Recordings involved subjective ratings (mood, physical fitness, fatigue) and objective measures (heart rate, vanillyl mandelic acid excretion, reaction times, critical flicker fusion, oral temperature). In addition, the subjects filled out questionnaires for personality traits (extroversion, neuroticis, anxiety) and behavioural characteristics (morningness, rigidity of sleeping habits, vigourness to overcome drowsiness). The volume of air traffic varied greatly with peaks during the day and low levels at night. Nevertheless, the heart rates of the group members showed quite constant levels in all the three shifts, irrespective of the workload. The same pattern appeared in the controllers' excretion of VMA, which remained high during both day and night shifts, regardless of the reduced workload. The subjective mood and physical fitness decreased similarly, while feelings of fatigue increased on all three shifts, particularly on the night shift. The circadian rhythm of the oral temperature showed a slight modification of the nocturnal depression during the night shift, caused by the state of awakeness and activity. However, the rhythm was not altered in its normal circadian phase, due to the fast shift rotation adopted. The psychophysiological responses were affected by personal characteristics, in particular morningness and ability to overcome drowsiness. PMID- 8404840 TI - Assessment of pilot performance and mental workload in rotary wing aircraft. AB - This research examined the processing demands imposed upon experienced pilots by two different communication formats, digital and verbal, in a high fidelity simulation of an advanced multi-function helicopter. The mental workload imposed by the type and magnitude of communications was assessed by a battery of subjective, performance, secondary, and physiological measures. The performance data indicated that the pilots had difficulty adhering to the Nap of the Earth altitude criterion with high communication demands, particularly with the digital communication system. This was presumably due to the requirement to spend more time scanning the multi-function displays with the digital than with the verbal communication system. On the other hand, the pilots were less prone to task shedding when they used the digital communication system possibly due to the provision of a permanent list of queries that was unavailable with the verbal system. Measures of heart rate variability and blink rate were larger with the verbal than with the digital system, presumably reflecting increased respiratory demands in the verbal condition as well as increased visual processing demands with the digital format. Finally, the probe evoked P300 component decreased in amplitude as a function of increases in the magnitude of communications. The results are discussed in terms of the structural and capacity demands of the communications systems that were proposed for the advanced multi-function helicopter. PMID- 8404841 TI - Comparing the concepts of mental load and stress. AB - This paper delineates mental load and stress as two related concepts that originate from different theoretical frameworks. A proper distinction between the two concepts is important, not only for theory building, but because it may lead also to different interpretations of experimental results, and, consequently, to different recommendations in applied situations. High workload is regarded as an important but not a critical factor in the development of stress symptoms. It is quite possible to work hard in difficult and complex tasks, even under unfavourable conditions, without cognitive strain, psychosomatic complaints, or adverse physiological effects. High task demands can be met by mobilizing extra energy through mental effort. This 'trying harder' reaction is a normal and healthy coping strategy to adapt to situational demands. In contrast, stress is regarded as a state in which the equilibrium between cognitive and energetical processes is disturbed by ineffective energy mobilization and negative emotions. Stress typically is characterized by inefficient behaviour, overreactivity, and the incapacity to recover from work. Stress is regarded as a state in which the physiological system is disorganized, which results in decreased well-being, sleeping problems, psychosomatic complaints, and increased health risks. PMID- 8404842 TI - Olive: a key gene required for chlorophyll biosynthesis in Antirrhinum majus. AB - Olive (oli) is a recessive nuclear mutation of Antirrhinum majus which reduces the level of chlorophyll pigmentation and affects the ultrastructure of chloroplasts. The oli-605 allele carries a Tam3 transposon insertion which has allowed the locus to be isolated. The oli gene encodes a large putative protein of 153 kDa which shows homology to the products of two bacterial genes necessary for tetrapyrrole-metal chelation during the synthesis of bacteriochlorophyll or cobyrinic acid. We therefore propose that the product of the oli gene is necessary for a key step of chlorophyll synthesis: the chelation of magnesium by protoporphyrin IX. Somatic reversion of the oli-605 allele produces chimeric plants which indicate that the oli gene functions cell-autonomously. Expression of oli is restricted to photosynthetic cells and repressed by light, suggesting that it may be involved in regulating the rate of chlorophyll synthesis in green tissues. PMID- 8404843 TI - Decoupling of photo- and proton cycle in the Asp85-->Glu mutant of bacteriorhodopsin. AB - Surface bound pH indicators were applied to study the proton transfer reactions in the mutant Asp85-->Glu of bacteriorhodopsin in the native membrane. The amino acid replacement induces a drastic acceleration of the overall rise of the M intermediate. Instead of following this acceleration, proton ejection to the extracellular membrane surface is not only two orders of magnitude slower than M formation, it is also delayed as compared with the wild-type. This demonstrates that Asp85 not only accepts the proton released by the Schiff's base but also regulates very efficiently proton transfer within the proton release chain. Furthermore, Asp85 might be the primary but is not the only proton acceptor/donor group in the release pathway. The Asp85-->Glu substitution also affects the proton reuptake reaction at the cytoplasmic side, although Asp85 is located in the proton release pathway. Proton uptake is slower in the mutant than in the wild-type and occurs during the lifetime of the O intermediate. This demonstrates a feed-back mechanism between Asp85 and the proton uptake pathway in bacteriorhodopsin. PMID- 8404844 TI - Residues within transmembrane segment M2 determine chloride conductance of glycine receptor homo- and hetero-oligomers. AB - We have expressed glycine receptor (GlyR) alpha and beta subunit cDNAs in HEK-293 cells to study the functional properties of homo- versus hetero-oligomeric GlyR channels. Dose-response curves of whole-cell currents in cells expressing alpha 1 subunits revealed an average Hill coefficient of h = 4.2. Co-expression with the beta subunit markedly increased glycine-gated whole-cell currents, which now exhibited a mean Hill coefficient of only h = 2.5. For alpha 1, alpha 2 and alpha 3 homo-oligomers, the main-state single-channel conductances were 86, 111 and 105 pS, respectively, recorded at symmetrical Cl- concentrations of 145 mM. The mutant alpha 1 G221A gave rise to a main-state of 107 pS. This indicates that the main-state of alpha homo-oligomers depends on residue 221 which is located within transmembrane segment M2. Importantly, the main-state conductances of alpha 1/beta, alpha 2/beta and alpha 3/beta hetero-oligomers were only 44, 54 and 48 pS, respectively. The latter values are similar to those found in spinal neurons, suggesting that native GlyRs are predominantly alpha/beta hetero-oligomers. Co expression of alpha 1 with mutant beta subunits revealed that residues within and close to segment M2 of the beta subunit determine the conductance differences between homo- and hetero-oligomers. PMID- 8404845 TI - Identification of Exo2 as the catalytic subunit of protein kinase A reveals a role for cyclic AMP in Ca(2+)-dependent exocytosis in chromaffin cells. AB - Digitonin-permeabilized chromaffin cells secrete catecholamines by exocytosis in response to micromolar Ca2+ concentrations, but lose the ability to secrete in response to Ca2+ as the cells lose soluble proteins through the plasma membrane pores. We have previously shown [Morgan and Burgoyne (1992) Nature, 355, 833-836] that cytosol can retard this loss of secretory competence and that two distinct stimulatory activities (Exo1 and Exo2) are present in cytosol. Here we report that Exo2 behaved as a single peak of activity through purification on hydroxyapatite, ammonium sulfate precipitation and gel filtration and the activity correlated with a single polypeptide of approximately 44 kDa on SDS gels. Protein sequencing of this band revealed it to be the catalytic subunit of cyclic AMP-dependent protein kinase (PKA). Both cyclic AMP and the commercially available catalytic subunit of PKA stimulated exocytosis in a dose-dependent manner which was absolutely dependent on the presence of micromolar Ca2+. These data show that PKA (Exo2) regulates Ca(2+)-dependent exocytosis in bovine adrenal chromaffin cells. PMID- 8404846 TI - SCAMP 37, a new marker within the general cell surface recycling system. AB - Secretory carrier membrane proteins (SCAMPs) are widely distributed as components of post-Golgi membranes that function as recycling carriers to the cell surface. In fibroblasts, SCAMPs are concentrated in compartments involved in the endocytosis and recycling of cell surface receptors while in neurons and other cell types having regulated transport pathways, SCAMPs are also components of regulated carriers (synaptic vesicles, secretion granules and transporter vesicles). Their presence in multiple pathways distinguishes them from proteins (e.g. recycling cell surface receptors and synaptic vesicle proteins) which are concentrated in selected pathways. The SCAMPs also do not appear to reside beyond the boundaries of these pathways. This distribution suggests that SCAMPs are general markers of membranes that function in cell surface recycling. The primary sequence of SCAMP 37 reveals a novel polypeptide containing a series of structural motifs, including a calcium binding domain, a leucine zipper and two zinc fingers. The very broad tissue distribution, subcellular localization and sequence analysis all predict that SCAMPs play a fundamental role in cell surface recycling. PMID- 8404847 TI - The heat shock cognate protein from Dictyostelium affects actin polymerization through interaction with the actin-binding protein cap32/34. AB - During isolation of the F-actin capping protein cap32/34 from Dictyostelium discoideum, a 70 kDa protein was copurified which by cloning and sequencing was identified as a heat shock cognate protein (hsc70). This protein exhibited a specific and MgATP-dependent interaction with the heterodimeric capping protein. To investigate the protein-protein interaction in vitro, we expressed all three polypeptides separately in Escherichia coli and performed reconstitution experiments of complete or truncated hsc70 with the 32 and 34 kDa subunits of the capping protein. Viscosity measurements and studies on the polymerization kinetics of pyrene-labeled actin showed that hsc70 increased the capping activity of cap32/34 up to 10-fold, whereas hsc70 alone had no effect on actin polymerization. In addition, hsc70 acted as a molecular chaperone by stimulating the refolding of the denatured 32 and 34 kDa subunits of the capping protein. To study the interaction of the two domains of hsc70 with cap32/34, the N-terminal 42 kDa ATPase region and the C-terminal 30 kDa tail of hsc70 were expressed separately in E. coli. The 32 and 34 kDa subunits were capable of associating with both domains of hsc70. The ATPase domain of hsc70, which is structurally related to actin, proved to be responsible for the increased capping activity of cap32/34, whereas the C-terminal tail of hsc70 was involved in folding of the subunits of cap32/34. Our data indicate a novel linkage between 70 kDa heat shock proteins and the actin cytoskeleton.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404848 TI - The metaphase II arrest in mouse oocytes is controlled through microtubule dependent destruction of cyclin B in the presence of CSF. AB - In unfertilized eggs from vertebrates, the cell cycle is arrested in metaphase of the second meiotic division (metaphase II) until fertilization or activation. Maintenance of the long-term meiotic metaphase arrest requires mechanisms preventing the destruction of the maturation promoting factor (MPF) and the migration of the chromosomes. In frog oocytes, arrest in metaphase II (M II) is achieved by cytostatic factor (CSF) that stabilizes MPF, a heterodimer formed of cdc2 kinase and cyclin. At the metaphase/anaphase transition, a rapid proteolysis of cyclin is associated with MPF inactivation. In Drosophila, oocytes are arrested in metaphase I (M I); however, only mechanical forces generated by the chiasmata seem to prevent chromosome separation. Thus, entirely different mechanisms may be involved in the meiotic arrests in various species. We report here that in mouse oocytes a CSF-like activity is involved in the M II arrest (as observed in hybrids composed of fragments of metaphase II-arrested oocytes and activated mitotic mouse oocytes) and that the high activity of MPF is maintained through a continuous equilibrium between cyclin B synthesis and degradation. In addition, the presence of an intact metaphase spindle is required for cyclin B degradation. Finally, MPF activity is preferentially associated with the spindle after bisection of the oocyte. Taken together, these observations suggest that the mechanism maintaining the metaphase arrest in mouse oocytes involves an equilibrium between cyclin synthesis and degradation, probably controlled by CSF, and which is also dependent upon the three-dimensional organization of the spindle. PMID- 8404850 TI - Eps8, a substrate for the epidermal growth factor receptor kinase, enhances EGF dependent mitogenic signals. AB - A method which allows direct cloning of intracellular substrates for receptor tyrosine kinases (RTKs) was developed. By applying this technique to the study of the epidermal growth factor receptor (EGFR) signaling pathway, we have isolated a cDNA, designated eps8, which predicts a approximately 92 kDa protein containing an SH3 domain. Eps8 also contains a putative nuclear targeting sequence. Antibodies specific to the eps8 gene product recognize a protein of M(r) 97 kDa and a minor 68 kDa component, which are closely related, as demonstrated by V8 proteolytic mapping. The product of the eps8 gene is tyrosine-phosphorylated in vivo following EGF stimulation of intact cells and associates with the EGFR, despite the lack of a functional SH2 domain. Several other RTKs are also able to phosphorylate p97eps8. Thus, the eps8 gene product represents a novel substrate for RTKs. Adoptive expression of the eps8 cDNA in fibroblastic or hematopoietic target cells expressing the EGFR resulted in increased mitogenic response to EGF, implicating the eps8 gene product in the control of mitogenic signals. PMID- 8404849 TI - Cognate gene clusters govern invasion of host epithelial cells by Salmonella typhimurium and Shigella flexneri. AB - The enteric pathogens Salmonella typhimurium and Shigella flexneri differ in most virulence attributes including infectivity, pathology and host range. We have identified a new assemblage of genes responsible for invasion properties of Salmonella which is remarkably similar in order, arrangement and sequence to the gene cluster controlling the presentation of surface antigens (spa) on the virulence plasmid of Shigella. In Salmonella, this chromosomally encoded complex consists of over 12 genes, mutations in which abolish bacterial entry into epithelial cells. Although these genera use distinct invasion antigens, a non invasive spa mutant of Salmonella could be rescued by the corresponding Shigella homolog. While spa promotes equivalent functions in Shigella and Salmonella, this constellation of genes has been acquired independently by each genus and displays motifs used by diverse antigen export systems including those required for flagellar assembly and protein secretion. PMID- 8404851 TI - Charged amino acids required for signal transduction by the m3 muscarinic acetylcholine receptor. AB - The five muscarinic acetylcholine receptor (mAChR) subtypes, termed m1-m5, transduce agonist signals across the plasma membrane by activating guanine nucleotide binding (G) proteins. The large cytoplasmic domain joining the fifth and sixth transmembrane segments of mAChRs plays a critical role in controlling the specificity of G protein coupling. In this study, we determined which sequences within this domain are required for activation of signaling by the m3 mAChR. By measuring the ability of normal and mutant m3 mAChRs to couple to the G protein pathway leading to activation of phospholipase C and Ca(2+)-dependent chloride currents in RNA-injected Xenopus oocytes, we found that two clusters of charged residues near the fifth and sixth transmembrane segments were required for normal signaling; furthermore, the position of these sequences was critical for their function. Finally, analysis of deletion mutant m3 mAChRs confirmed the importance of these sequences; receptors containing as few as 22 out of 239 amino acids of the cytoplasmic domain were fully active in signaling if they included the critical charged residues. Sequence comparisons suggest that similar charged sequences may be required for signal transduction by many G protein-coupled receptors. PMID- 8404852 TI - Phosphorylation of KSP motifs in the C-terminal region of titin in differentiating myoblasts. AB - Titin is a giant structural protein of striated muscle (M(r) approximately 3000 kDa) and single molecules span sarcomeres from the M- to Z-lines. We have cloned and sequenced the C-terminal region of the titin molecule, which is an integral part of M-lines and forms intimate contacts with the 165 and 190 kDa M-line proteins. In contrast to the regular motif patterns of the A-band portion of titin, the 5.7 kb of titin sequences from the M-line show a complex structure of immunoglobulin-C2 repeats, separated by unique interdomain insertion sequences. As a striking feature, one interdomain insertion comprises four KSP repeats analogous to the multi-phosphorylation repeats of neurofilament subunits H and M. In vitro phosphorylation assays with expressed titin KSP sequences detect high levels of titin KSP phosphorylating kinases in developing but not in differentiated muscle. Since this kinase activity can be depleted from myocyte extracts by antibodies against cdc2 kinase and p13suc1 beads, the titin KSP kinase is structurally related to cdc2 kinase. We suggest that titin C-terminal phosphorylation by SP-specific kinases is regulated during differentiation, and that this may control the assembly of M-line proteins into regular structures during myogenesis. PMID- 8404853 TI - Sox-4, an Sry-like HMG box protein, is a transcriptional activator in lymphocytes. AB - Previous studies in lymphocytes have described two DNA-binding HMG box proteins, TCF-1 and LEF-1, with affinity for the A/TA/TCAAAG motif found in several T cell specific enhancers. Evaluation of cotransfection experiments in non-T cells and the observed inactivity of an AACAAAG concatamer in the TCF-1/LEF-1-expressing T cell line BW5147, led us to conclude that these two proteins did not mediate the observed enhancer effect. We therefore searched for additional HMG box proteins. By a PCR-aided strategy, we cloned Sox-4, a gene with homology to the HMG box region of the sex determining gene SRY. Sox-4 was expressed in T and pre-B lymphocyte lines and in the murine thymus. Significantly, BW5147 T cells did not express Sox-4. Recombinant Sox-4 bound with high affinity (Kd 3 x 10(-11) M) to the minor groove of the AACAAAG motif, most likely contacting all seven base pairs. In contrast with observations on TCF-1 and LEF-1, cotransfection with Sox 4 unveiled a transactivating capacity, which mapped to its serine-rich C terminus. This region remained functional upon grafting onto a GAL4 DNA-binding domain. Sox-4 is thus the first 'classical' transcription factor in the Sox gene family with separable DNA-binding and transactivation domains. Our observations indicate that a detailed understanding of T cell-specific gene control must integrate the concerted activity of at least three tissue-specific HMG box genes. PMID- 8404854 TI - Deposition of chromosomal protein HMG-17 during replication affects the nucleosomal ladder and transcriptional potential of nascent chromatin. AB - A cell-free system from Xenopus eggs was used to study the role of chromosomal protein HMG-17 in the generation of the chromatin structure of transcriptionally active genes. Addition of HMG-17 protein to the extracts, which do not contain structural homologs of the HMG-14/-17 protein family, indicates the protein is incorporated into the nascent template during replication, prior to completion of chromatin assembly. The protein binds to and stabilizes the structure of the nucleosomal core thereby improving the apparent periodicity of the nucleosomal spacing of nascent chromatin. Assembly of HMG-17 into the nascent chromatin structure significantly increased the transcription potential of the 5S RNA gene and satellite I chromatin. Kinetic studies indicate that the increase in transcriptional potential is observed only when HMG-17 is incorporated into nucleosomes during chromatin assembly. PMID- 8404855 TI - Molecular mechanisms of pattern formation by the BRE enhancer of the Ubx gene. AB - The core activity of the Ubx gene enhancer BRE (bx region enhancer) is encoded within a 500 bp module. bx DNA outside this active module increases the level of expression, expands the expression into ventro-lateral ectoderm and partially stabilizes the late expression pattern. The products of the gap genes hb and tll and of the pair-rule gene ftz bind to the 500 bp BRE module and control directly its initial pattern of expression. ftz enhances expression in even-numbered parasegments within the correct spatial domain whose boundaries are set by hb and tll. In addition, en and twi products activate the enhancer, probably directly. en broadens the parasegmental stripe while twi cooperates with ftz to enhance expression in the mesoderm. Binding sites for the five regulators are closely clustered, often overlapping extensively with one another. In vitro, hb blocks the binding of ftz and can also displace ftz protein pre-bound to an overlapping site, suggesting that competitive binding and/or interference by hb sets the initial boundaries of the domain of expression. Our results also suggest that this interaction is short-range and the long distance interactions among different enhancers may depend on each enhancer's ability to complex with the promoter. PMID- 8404856 TI - Cross-coupling of the NF-kappa B p65 and Fos/Jun transcription factors produces potentiated biological function. AB - NF-kappa B and AP-1 represent distinct mammalian transcription factors that target unique DNA enhancer elements. The heterodimeric NF-kappa B complex is typically composed of two DNA binding subunits, NF-kappa B p50 and NF-kappa B p65, which share structural homology with the c-rel proto-oncogene product. Similarly, the AP-1 transcription factor complex is comprised of dimers of the c fos and c-jun proto-oncogene products or of closely related proteins. We now demonstrate that the bZIP regions of c-Fos and c-Jun are capable of physically interacting with NF-kappa B p65 through the Rel homology domain. This complex of NF-kappa B p65 and Jun or Fos exhibits enhanced DNA binding and biological function via both the kappa B and AP-1 response elements including synergistic activation of the 5' long terminal repeat of the human immunodeficiency virus type 1. These findings support a combinatorial mechanism of gene regulation involving the unexpected cross-coupling of two different classes of transcription factors to form novel protein complexes exhibiting potentiated biological activity. PMID- 8404857 TI - The oncoprotein Bcl-3 can facilitate NF-kappa B-mediated transactivation by removing inhibiting p50 homodimers from select kappa B sites. AB - Previously we have proposed a role for Bcl-3 in facilitating transactivation through kappa B sites by counteracting the inhibitory effects of bound, non transactivating homodimers of the p50 subunit of NF-kappa B. Such homodimers are abundant for example in nuclei of unstimulated primary T cells. Here we extend the model and provide new evidence which fulfills a number of predictions. (i) Bcl-3 preferentially targets p50 homodimers over NF-kappa B heterodimers since the homodimers are completely dissociated from kappa B sites at concentrations of Bcl-3 which do not affect NF-kappa B. (ii) Select kappa B sites associate very strongly and stably with p50 homodimers, completely preventing binding by NF kappa B. Such kappa B sites are likely candidates for regulation by p50 homodimers and Bcl-3. (iii) Bcl-3 and p50 can be co-localized in the nucleus, a requirement for active removal of homodimers from their binding sites in vivo. (iv) The ankyrin repeat domain of Bcl-3 is sufficient for the reversal of p50 homodimer-mediated inhibition, correlating with the ability of this domain alone to inhibit p50 binding to kappa B sites in vitro. Our data support the model that induction of nuclear Bcl-3 may be required during cellular stimulation to actively remove stably bound p50 homodimers from certain kappa B sites in order to allow transactivating NF-kappa B complexes to engage. This exact mechanism is demonstrated with in vitro experiments. PMID- 8404858 TI - Multiple and cooperative phosphorylation events regulate the CREM activator function. AB - Phosphorylation is one of the major mechanisms by which the activity of transcription factors can be regulated. We have investigated the role of phosphorylation in the regulation of the transcription factor CREM. We show that the CREM tau activator is phosphorylated on multiple serine and threonine residues in vivo. Stimulation of various signal transduction pathways by forskolin, TPA or Ca2+ ionophore leads to enhanced phosphorylation of serine 117, concomitant with an increase in the transactivation potential of CREM tau. We have identified multiple kinases that can also phosphorylate S117 in vitro. Moreover, we show that casein kinase I and II cooperatively phosphorylate CREM tau on multiple residues, eliciting enhanced DNA binding. Cooperative phosphorylation is also observed with other kinases. These results show that the activity of CREM tau is regulated by multiple phosphorylation events, suggesting that CREM could be considered as a nuclear effector where signalling pathways may converge and/or cross-talk. PMID- 8404859 TI - Regulatory elements in the immunoglobulin kappa locus induce c-myc activation and the promoter shift in Burkitt's lymphoma cells. AB - In Burkitt's lymphoma cells the proto-oncogene c-myc is constantly juxtaposed through chromosomal translocation to one of the immunoglobulin loci on chromosomes 14, 2 or 22. In the majority of cases the chromosomal breakpoint is localized 3' or 5' of the gene leaving the physiological c-myc transcription unit intact. As a consequence of the translocation the c-myc gene on the translocation chromosome becomes transcriptionally activated in such a manner that the c-myc promoter P1 is more active than promoter P2. In order to define elements involved in c-myc activation through t(2;8) translocation we have studied the expression of constructs consisting of c-myc and different parts of the immunoglobulin kappa locus after stable transfection into Burkitt's lymphoma cells. The c-myc gene under the control of the complete Ig kappa locus containing matrix attachment region, intron enhancer, constant kappa gene and 3' enhancer was strongly activated with predominant usage of promoter P1. Deletion analysis revealed that the intron or 3' enhancers alone activated c-myc to a much lesser extent and with normal promoter usage (P1 < P2). The cooperation of the same regulatory elements is required not only for transcriptional activation and induction of the promoter shift but also for down-regulation of promoter P1 of the translocated c-myc allele by sodium butyrate, another characteristic feature of Burkitt's lymphoma cells. This supports the notion that all elements involved in transcriptional activation and dysregulation of c-myc are contained within the myc-Ig specific minichromosome. PMID- 8404860 TI - A transcription factor with homology to the AP-1 family links RNA transcription and DNA replication in the lytic cycle of Epstein-Barr virus. AB - oriLyt, the lytic origin of DNA replication of Epstein-Barr virus (EBV), ensures viral DNA amplification during the productive or lytic phase of the virus' life cycle. To understand the contribution of cis- and transacting elements involved in DNA replication of oriLyt, a detailed mutational analysis was undertaken which defined BZLF1, a viral transcriptional activator, as an essential replication factor. The BZLF1 protein belongs to the extended fos/jun family of transcription factors and binds to specific BZLF1 binding motifs within oriLyt, as well as to consensus AP-1 sites. Recombinant, chimeric transcription factors identified the transcriptional activation domain of BZLF1 as being necessary to mediate DNA replication, a function which could not be substituted by any other transcription factor tested, including jun, E2, myc or VP16. PMID- 8404861 TI - A critical role for heat shock transcription factor in establishing a nucleosome free region over the TATA-initiation site of the yeast HSP82 heat shock gene. AB - Heat shock genes are poised for rapid transcriptional activation in response to environmental stress. A universal structural characteristic of such genes is the presence of a nucleosome-free, DNase I hypersensitive promoter region. Here we investigate the structural and functional effects of mutating HSE1, the preferred heat shock factor (HSF) binding site upstream of the yeast HSP82 gene. In situ deletion or substitution of this sequence reduces both basal and induced transcription by at least two orders of magnitude. Moreover, such mutations lead to a dramatic transition in chromatin structure: the DNase I hypersensitive region is replaced by two stable, sequence-positioned nucleosomes. One of these is centered over the mutated heat shock element, while the other--as revealed by DNase I genomic footprinting--is precisely positioned in a rotational sense over the TATA-initiation site. Overexpression of yeast HSF strongly suppresses the null phenotype of the induced hsp82-delta HSE1 gene and re-establishes DNase I hypersensitivity over its promoter. Such suppression is mediated through sequence disposed immediately upstream of HSE1 and containing two low affinity heat shock elements. These data imply a critical role for HSF in displacing stably positioned nucleosomes in Saccharomyces cerevisiae and suggest that HSF transcriptionally activates HSP82 at least partly through its ability to alleviate nucleosome repression of the core promoter. PMID- 8404862 TI - pH regulation is a major determinant in expression of a fungal penicillin biosynthetic gene. AB - Transcription of the ipnA gene encoding isopenicillin N synthetase, an enzyme of secondary metabolism, is under the control of the pH regulatory system in the fungus Aspergillus nidulans. External alkaline pH or mutations in pacC, the wide domain regulatory gene which mediates pH regulation, override carbon regulation of ipnA transcript levels, resulting in elevation of the levels of this message in sucrose broth. Strains carrying these mutations, which mimic growth at alkaline pH, produce higher levels of penicillins when grown in sucrose broth compared with the wild type strain grown under carbon derepressing conditions. ipnA transcription is regulated by carbon (C) source, but extreme mutations in creA (the regulatory gene mediating carbon catabolite repression) only slightly increase repressed transcript levels. Precise deletion of the only in vitro CreA binding site present in a region of the ipnA promoter involved in carbon regulation has no effect on ipnA expression. The levels of ipnA transcript in broths with acetate or glycerol as principal C sources are inconsistent with direct or indirect creA-mediated transcriptional control of the gene. We conclude that a second, creA-independent mechanism of carbon repression controls expression of this gene. All derepressing C sources tested result in alkalinization of the growth media. In contrast, all repressing C sources result in external acidification. Neither acidic external pH nor pal mutations, mimicking the effects of growth at acid pH, prevent carbon derepression, providing strong support for independent regulatory mechanisms, one mediating carbon regulation (via thus far unidentified genes) and another mediating pH regulation (through the pacC-encoded transcriptional regulator). External pH measurements suggest that these two independent forms of regulation normally act in concert. We propose that external alkalinity represents a physiological signal which triggers penicillin biosynthesis. PMID- 8404863 TI - Transcription of ftsZ oscillates during the cell cycle of Escherichia coli. AB - The FtsZ protein is a key element controlling cell division in Escherichia coli. A powerful transcription titration assay was used to quantify the ftsZ mRNA present in synchronously dividing cells. The ftsZ mRNA levels oscillate during the cell cycle reaching a maximum at about the time DNA replication initiates. This cell cycle dependency is specifically due to the two proximal ftsZ promoters. A strain was constructed in which expression of ftsZ could be modulated by an exogenous inducer. In this strain cell size and cell division frequency were sensitive to the cellular FtsZ contents, demonstrating the rate limiting role of this protein in cell division. Transcriptional activity of the ftsZ promoters was found to be independent of DnaA, indicating that DNA replication and cell division may be independently controlled at the time when new rounds of DNA replication are initiated. This suggests a parallelism between the prokaryotic cell cycle signals and the START point of eukaryotic cell cycles. PMID- 8404864 TI - Mammalian ribonucleotide reductase R1 mRNA stability under normal and phorbol ester stimulating conditions: involvement of a cis-trans interaction at the 3' untranslated region. AB - Ribonucleotide reductase R1 gene expression is elevated in BALB/c 3T3 fibroblasts treated with the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA). We show that TPA treatment increased the half-life of R1 mRNA by 5-fold, indicating that TPA regulates R1 gene expression by a post-transcriptional mechanism. We investigated the possibility that the 3' untranslated region (3'UTR) of R1 mRNA contains regulatory information for TPA-mediated message stability. Our studies demonstrated that a 49 nucleotide (nt) TPA-responsive region existed within the R1 mRNA 3'UTR. Deletion of the 49 nt region led to the abolishment of TPA-induced stability of R1 and hybrid CAT mRNAs. Further deletions of the 3'UTR did not significantly affect mRNA turnover rates. In addition, we detected by cross linking a novel 52-57 kDa R1 mRNA-binding protein (R1BP) that bound selectively to the 49 nt region of the R1 mRNA 3'UTR and did not bind to the 5'UTR, the coding region or other mRNA 3'UTRs. The R1BP-RNA binding activity observed in unstimulated cells was rapidly and markedly down-regulated after TPA treatment, suggesting a role for R1BP in mRNA metabolism, and in the mechanism of action of TPA-induced R1 message stabilization. These results support a novel model of R1 gene regulation in which a cis-element(s) within the 49 nt region of the R1 mRNA 3'UTR interacts with R1BP in a mechanism that regulates R1 message stability. We propose that this model accounts for the TPA-mediated stability alteration of R1 message, through down-regulation of R1BP-RNA binding activity linked to a reduction in the rate of R1 mRNA degradation. PMID- 8404865 TI - A Saccharomyces cerevisiae homologue of mammalian translation initiation factor 4B contributes to RNA helicase activity. AB - The TIF3 gene of Saccharomyces cerevisiae was cloned and sequenced. The deduced amino acid sequence shows 26% identity with the sequence of mammalian translation initiation factor eIF-4B. The TIF3 gene is not essential for growth; however, its disruption results in a slow growth and cold-sensitive phenotype. In vitro translation of total yeast RNA in an extract from a TIF3 gene-disrupted strain is reduced compared with a wild-type extract. The translational defect is more pronounced at lower temperatures and can be corrected by the addition of wild type extract or mammalian eIF-4B, but not by addition of mutant extract. In vivo translation of beta-galactosidase reporter mRNA with varying degree of RNA secondary structure in the 5' leader region in a TIF3 gene-disrupted strain shows preferential inhibition of translation of mRNA with more stable secondary structure. This indicates that Tif3 protein is an RNA helicase or contributes to RNA helicase activity in vivo. PMID- 8404866 TI - A new yeast translation initiation factor suppresses a mutation in the eIF-4A RNA helicase. AB - We have isolated a gene, STM1, which encodes a new translation initiation factor from Saccharomyces cerevisiae. The gene acts, if present on a multicopy plasmid, as a suppressor of a temperature-sensitive mutation in eIF-4A. The single copy STM1 gene is not essential, but disruption causes a slow growth phenotype. Analysis of polysomes from a strain carrying a disrupted stm1 allele shows a clear defect in translation initiation as shown by a strong reduction in polysomes and an increase in the monosomes. Sequence analysis revealed interesting features of the putative Stm1 protein. Comparison of the entire protein sequence with databanks showed some similarity with the human eIF-4B protein. The Stm1 protein has potential RNP1 and RNP2 motifs characteristic for RNA-binding proteins. The protein also contains six highly conserved direct repeats of 21-26 amino acids and one partial repeat. PMID- 8404867 TI - Mammalian polypeptide chain release factor and tryptophanyl-tRNA synthetase are distinct proteins. AB - A very high (approximately 90%) structural similarity exists between the bovine, human and murine tryptophanyl-tRNA synthetases (WRS), and quite unexpectedly the rabbit polypeptide chain release factor (eRF). This similarity may point to a very close resemblance or identity between these proteins involved in distinct steps of protein synthesis, or inadvertently to an incorrect assignment of the clone reported to encode eRF, since the structure of clones encoding WRS were confirmed by peptide sequencing. Using high resolution column chromatography and sucrose gradient centrifugation combined with assays for WRS and eRF activities, we show that functionally distinct WRS and eRF proteins can be completely separated from each other. Moreover, a putative anti-eRF monoclonal antibody appears incapable of immunoprecipitating the eRF activity or binding to protein(s) possessing eRF activity. This antibody binds to protein fractions which coincide in various separation procedures with rabbit WRS activity, and to pure bovine WRS. The protein expressed in Escherichia coli from the original cDNA clone initially reported to encode eRF, has WRS activity but not eRF activity. Resequencing of the fragment of the original rabbit cDNA demonstrates the presence of the previously overlooked HXGH motif typical of class I aminoacyl tRNA synthetases. Consequently, mammalian WRS and eRF are different proteins, and the cDNA clone formerly assigned as encoding eRF encodes rabbit WRS. PMID- 8404868 TI - Degradation of Mos by the N-terminal proline (Pro2)-dependent ubiquitin pathway on fertilization of Xenopus eggs: possible significance of natural selection for Pro2 in Mos. AB - The c-mos proto-oncogene product (Mos), an essential component of the cytostatic factor responsible for meiotic arrest in vertebrate eggs, undergoes specific proteolysis soon after fertilization or activation of Xenopus eggs. To determine the degradation pathway of Mos on egg activation, various Mos mutants were expressed in Xenopus eggs and their degradation on egg activation was examined. Mos degradation absolutely required its penultimate proline (Pro2) residue and dephosphorylation of the adjacent serine (Ser3) residue. These degradation signals were essentially the same as those of Mos in meiosis I of Xenopus oocyte maturation, where Mos has been shown to be degraded by the 'second-codon rule' based ubiquitin pathway. To test whether Mos degradation on egg activation is also mediated by the ubiquitin pathway, we attempted to identify and abrogate a specific ubiquitination site(s) in Mos. We show that the major ubiquitination site in Mos is a Lys34 residue and that replacement of this residue with a non ubiquitinatable Arg residue markedly enhances the stability of Mos on egg activation. These results indicate that the degradation of Mos on egg activation or fertilization is mediated primarily by the N-terminal Pro2-dependent ubiquitin pathway, as in meiosis I of oocyte maturation. The N-terminal Pro2 residue of Mos appears to be naturally selected primarily for its degradation on fertilization, rather than that in meiosis I. PMID- 8404869 TI - Alternating d(GA)n DNA sequences form antiparallel stranded homoduplexes stabilized by the formation of G.A base pairs. AB - Alternating d(GA)n DNA sequences form antiparallel stranded homoduplexes which are stabilized by the formation of G.A pairs. Three base pairings are known to occur between adenine and guanine: AH+ (anti).G(syn), A(anti).G(anti) and A(syn).G(anti). Protonation of the adenine residues is not involved in the stabilization of this structure, since it is observed at any pH value from 8.3 to 4.5; at pH < or = 4.0 antiparallel stranded d(GA.GA) DNA is destabilized. The results reported in this paper strongly suggest that antiparallel stranded d(GA.GA) homoduplexes are stabilized by the formation of alternating A(anti).G(anti) and G(anti).A(syn) pairs. In this structure, all guanine residues are in the anti conformation with their N7 position freely accessible to DMS methylation. On the other hand, adenines in one strand adopt the anti conformation, with their N7 position also free for reaction, while those of the opposite strand are in the syn conformation, with their N7 position hydrogen bonded to the guanine N1 group of the opposite strand. A regular right-handed helix can be generated using alternating G(anti).A(syn) and A(anti).G(anti) pairs. PMID- 8404870 TI - The td intron endonuclease I-TevI makes extensive sequence-tolerant contacts across the minor groove of its DNA target. PMID- 8404871 TI - Big Blue transgenic mouse lacI mutation analysis. PMID- 8404872 TI - Use of fluorescence in situ hybridization to detect chromosome-specific changes in exfoliated human bladder and oral mucosa cells. AB - Change in chromosome number, numerical aneuploidy, has been consistently linked with cancer development. Since 90% of cancers arise in epithelial tissues, techniques that measure aneuploidy in these tissues would be very useful. Here we describe methods of optimization and suggest use of fluorescent in situ hybridization (FISH) to detect aneuploidy in exfoliated epithelial cells collected from the mouth and bladder. A total of 10,383 urothelial cells and 4,691 buccal cells were scored in order to determine a baseline frequency of aneuploidy in human volunteers using a classical satellite probe for chromosome 9. Protein digestion with pepsin was found to be more efficient at removing the keratinized cell membrane and optimizing probe penetration than acid washes, detergent washes, or hypotonic treatments. A 20 min cellular digestion with 200 micrograms/ml and a 30 min digestion with 300 micrograms/ml of pepsin in 0.01 M HCl optimized probe penetration in urothelial and buccal cells, respectively. Average frequencies for 0, 1, 2, 3, and 4 hybridization regions were 10.3, 10.1, 78.4, 1.0, and 0.3% for urothelial cells and 8.8, 9.8, 79.4, 1.3, and 0.3% for buccal cells, respectively. These results are very similar to those previously described in lymphocytes. The urothelial cells of males had a lower frequency of diploid cells and a higher frequency of cells without hybridization regions than females (P < 0.02). No statistically significant variability was found between individuals or sex groups in buccal cells. Our data show that FISH is a useful tool to detect changes in frequency of aneuploidy in exfoliated epithelial cells and has good potential for monitoring human populations exposed to genotoxic agents. PMID- 8404873 TI - Spectrum of spontaneous HPRT- mutations in TK6 human lymphoblasts. AB - The occurrence of deletions, coding sequence alterations, and intronic changes leading to aberrant splicing has been characterized among 33 spontaneous HPRT- mutants in TK6 human lymphoblasts. Deletions detectable by multiplex PCR amplification accounted for 45% (15/33) of the mutant collection. Base substitutions represented 30% (10/33) of the total, and were predominated by changes at G:C base pairs. The remaining mutants were distributed among frameshifts (9%, 3/33), small deletions (6%, 2/33), and compound alterations (9%, 3/33). Five mutants (15%) demonstrated aberrant splicing of the hprt transcript. A cluster of 4 deletion/insertion events was identified in hprt exon 6. A nearly perfect 13 bp duplication differed from the original sequence only by an A:T to G:C transition, which was observed as a unique alteration in another HPRT- mutant. A model involving correction of a mismatch in a secondary structure formed by the duplicated sequence may account for these results. PMID- 8404874 TI - Induction of sister chromatid exchange in spleen and bone marrow cells of rats exposed by inhalation to different dose rates of ethylene oxide. AB - We investigated the effects of dose rate on the frequency of sister chromatid exchange (SCE) in bone marrow and spleen cells of rats exposed to ethylene oxide (EtO). Four groups (18/group) of male Fischer 344 rats were exposed to EtO by inhalation. The exposures consisted of 100 ppm for 6 hr/day, 300 ppm for 2 hr/day, 600 ppm for 1 hr/day, and clean air control. All EtO treated rats were given a total exposure dose of 600 ppm.hr daily, 5 days/week for 3, 6, or 9 months. Six rats per group were sacrificed at each time point, and SCEs were measured in cultured spleen and bone marrow cells. A statistically significant increase was found in SCEs in both bone marrow and spleen cells for all treated groups and at each time point when compared to the control, except at the 3-month exposure for the middle and high dose-rate groups in bone marrow cells. In the spleen, the increases in SCEs were similar among the three experimental groups. In bone marrow, the lowest dose rate (100 ppm) resulted in higher SCE frequencies than the medium and high dose-rate group after 3 and 6 month exposures. The overall frequencies of SCEs in the spleen cells were higher than in the bone marrow cells. The increase in SCE frequencies and decrease in the replicative index in spleen cells were also dependent on the duration of exposure. These results indicate that (1) EtO, by inhalation, can cause SCEs both in spleen and bone marrow cells of Fischer 344 rats, (2) spleen cells are more sensitive to EtO than bone marrow cells, and (3) in bone marrow cells the lowest dose-rate (longest) exposure causes more SCEs than the highest dose-rate (shortest) exposures. PMID- 8404875 TI - Modulation of the H2O2-induced SOS response in Escherichia coli PQ300 by amino acids, metal chelators, antioxidants, and scavengers of reactive oxygen species. AB - The SOS chromotest is a simple colorimetric genotoxicity assay that monitors DNA repair by measuring the induction of the gene sfiA in Escherichia coli K-12. E. coli PQ300, a diagnostic SOS tester strain for the detection of oxidative genotoxins, carries a mutation in a key gene for antioxidative defense, oxyR. This mutation renders PQ300 more sensitive to oxidative genotoxins, particularly to H2O2. We found that induction of the SOS response by H2O2 in E. coli PQ300 is dependent on the composition of the incubation medium; a substantially reduced response was obtained in minimal phosphate buffered saline (PBS) as opposed to complex Luria broth (LB) medium. Supplementation of PBS with histidine or cysteine stimulated H2O2-induced SOS induction to levels exceeding those found in LB medium. Low concentrations of glutathione (20-70 microM) also enhanced the H2O2-induced SOS response in E. coli PQ300, whereas higher concentrations (> 150 microM) were protective. Preincubation of tester cells with the chelators o phenanthroline, 2,2-dipyridyl, and ethylenediaminetetraacetic acid (EDTA) protected cells from the effects of H2O2, although EDTA was only partially effective. Pretreatment of PQ300 with the antioxidant ascorbic acid or the hydroxyl radical scavenger dimethyl sulfoxide also diminished the SOS response, whereas mannitol and glucose were ineffective. The results show that the net effect of H2O2-induced DNA damage is influenced by the balance of oxidative and antioxidative factors and, furthermore, can be modulated by constituents of the extracellular milieu. PMID- 8404876 TI - Antimutagenic potency of chlorophyllin in the Salmonella assay and its correlation with binding constants of mutagen-inhibitor complexes. AB - Chlorophyllin (CHL) is a water-soluble salt of chlorophyll that exhibits antimutagenic activity in short-term genotoxicity assays and inhibits carcinogen DNA binding in vivo. The antimutagenic potency of CHL was studied against several structurally related heterocyclic amines using the Salmonella assay. The mutagens included 2-amino-3-methylimidazo[4,5,-f]-quinoline (IQ) and seven related IQ-type compounds, and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) and three additional non-IQ-type compounds. No relationship was observed between mutagenic potency (revertants/ng mutagen) and antimutagenic potency when expressed in terms of the CHL dose/plate-inhibiting mutagenicity by 50 percent (I50). However, a correlation was observed between mutagenic potency and the mole ratio of CHL to mutagen giving 50% inhibition (MR50), with most mutagens requiring several hundredfold to several thousandfold molar excess of CHL for inhibition. In spectrophotometric studies, CHL formed noncovalent molecular complexes with the heterocyclic amines, with binding constants in the range 3-13 x 10(3) M-1. Binding constants were inversely correlated with I50 and MR50 values, i.e., with increasing strength of complex formation less CHL/plate and a lower mole ratio of CHL to mutagen was required to inhibit mutagenicity. The results support an inhibitory mechanism in which chlorophylls operate as "interceptor molecules," interacting with carcinogens and mutagens directly and limiting their bioavailability. PMID- 8404877 TI - Comparative studies on cytotoxic and genotoxic effects of two organic mercury compounds in lymphocytes and gastric mucosa cells of Sprague-Dawley rats. AB - Human lymphocytes (HL) as well as lymphocytes (RL), hepatocytes (RH), and gastric mucosa cells (GM) of Sprague-Dawley rats were treated in vitro for 1 h with methylmercury chloride (MMC, 0.5-4 micrograms/ml) and dimethylmercury (DMM, 5-40 micrograms/ml). The cytotoxicity of the two organic mercury compounds was assessed by dye exclusion, and the extent of induced DNA fragmentation was measured with a single-cell microgel electrophoresis assay. Both MMC and DMM induced DNA damage and cytotoxicity in a dose-related manner in HL, RL, and GM. MMC was more effective in causing a significant increase in median DNA migration than DMM at doses yielding approximately the same degree of cytotoxicity. In rat hepatocytes the MMC-induced DNA damage was, however, lower than in the other cells. An analysis of repair kinetics following exposure to 2 micrograms/ml MMC was carried out in human lymphocytes obtained from an adult male donor. The bulk of DNA repair occurred 90 min after in vitro exposure, and it was about complete by 120 min following cessation of exposure. Finally, in order to have a basis for extrapolating to the human situation, in vivo studies were performed with Sprague Dawley rats, also assessing the DNA damage and cytotoxicity in the lymphocytes and gastric mucosa cells. These in vivo results after oral exposure may be directly compared to the in vitro data obtained in the same cells. PMID- 8404878 TI - Test of chiral recognition in the Salmonella typhimurium (TA100) mutagenicity of mucochloric acid-cysteine adducts. AB - A difference in biological response to enantiomers is not an uncommon observation and is, therefore, to be expected in various manifestations of genotoxicity. The bacterial mutagen mucochloric acid (2,3-dichloro-5-hydroxy-2(5H)-furanone) has one chiral center, at C-5, but this mutagen exists in racemic form because of the facile stereoisomerization occurring by the mechanism of ring-chain tautomerism. Two readily synthesized enantiomeric analogs of mucochloric acid, as well as the racemic form of the two, were prepared from mucochloric acid and (R)-(+)-, (S)-( )-, and (R,S)-(+/-)-cysteine. Using Salmonella typhimurium (TA100), the enantiomeric compounds were assayed together in four dose/response assays along with mucochloric acid, the reference mutagen. In three of the same four assays, the racemic form was also assayed. Neither statistically significant differences in mutagenicity, as determined in slope responses, nor distinctions from the plotted curves were observed among the two enantiomers and their racemic form. Therefore, no enantiospecific interaction between enantiomers and chiral DNA or enzymes involved in repair or replication could be concluded. PMID- 8404879 TI - Screening of azo dyes for mutagenicity with Ames/Salmonella assay. AB - Azo dyes, the largest portion of manufactured dyestuffs, are primarily used as colouring substances in food, textiles, and the plastic industry. It has been estimated that 128 tonnes per annum of dyes are released into the environment worldwide [Anliker, 1977]. Certain azo compounds are known to be mutagenic in bacterial tests [Yahagi et al., 1975; Venitt and Bushell, 1976; Brown et al., 1978]. Watersoluble dyes are biotransformed by intestinal micro-organisms in the gastro intestinal tract, and the toxicity, mutagenicity, and carcinogenicity of these dyes in the gut or liver may be attributed to their metabolites. Since it is desirable to have a genotoxic evaluation of a chemical being released into the environment in order to check their indiscriminate use, a project has been initiated to determine the mutagenicity of the azo dyes being used commercially. The present report deals with the results of 13 dyes tested in Salmonella typhimurium with and without metabolic activation. PMID- 8404880 TI - Predicting chemical carcinogenesis in rodents: commentary on the results of a workshop. PMID- 8404881 TI - A six month prospective study on different aspects of abortion. AB - A six month prospective study on various aspects of abortion was conducted from April 1, 1991 to Sept. 30, 1991 in Sidamo Regional Hospital (Yirgalem). A total of 185 cases of abortion were seen. Of these, 64 (35%) were induced and 121 (65%) were spontaneous. There were 2 deaths in the illegally induced group abortion, 1 death in the other group. Induced abortion was higher in age group 20-24 (61%), single (65%), unemployed (70%), nulliparous (48%) and 7-12 grade educational level (67%). The pregnancy was unwanted in all cases of induced abortion and in 50 (41%) of the spontaneous cases. The common instruments used for inducing abortion were plastic catheters (58%) and metallic instruments (32%). The abortionists were mainly health workers (55%). The mean hospital stay was 6.3 days for illegally induced and 2.1 days for spontaneous abortions. The incidence of septic abortion was found to be statistically significantly higher in induced than in spontaneous abortion (p < 0.001) while haemorrhagic shock was not (p > 0.05). The type of anaesthesia and required procedure are also analyzed in this study. Eighty-eight percent of the study population did not use any type of contraception. The role of contraception in preventing unwanted pregnancy and therefore induced abortion is discussed. PMID- 8404882 TI - Hepatitis E virus infection in pregnancy in Ethiopia. AB - Thirty-two pregnant and 34 nonpregnant Ethiopian women between 15 and 45 years of age with sporadic acute viral hepatitis were studied consecutively. Demographic data including family size, monthly income and nutritional status as well as hepatitis virus markers were compared in the pregnant and nonpregnant groups. Only 3 nonpregnant women had hepatitis A infection. Hepatitis B infection was diagnosed in 4 pregnant and 9 nonpregnant women. Nineteen (59%) pregnant women had hepatitis E virus (HEV) infection as compared to 7 (22%) in the nonpregnant group (Relative risk = 2.88; 95% Confidence interval = 1.4-5.9). The remaining 9 pregnant and 15 nonpregnant women had non-A, non-B, non-E (NANBNE) hepatitis. Of a total of 10 maternal deaths, 9 occurred (7 during the third trimester) in the pregnant group, 8 in association with HEV infection. Two deaths, one from each group, were due to NANBNE hepatitis. In addition to 6 foetal losses as a result of maternal death, there were 2 foetal deaths and 7 premature deliveries as a direct result of acute viral hepatitis, all but 2 associated with HEV infection. Comparison of socioeconomic and nutritional status, clinical features, mean aminotransferase and bilirubin levels did not show differences in the two groups. Thus, pregnant women are more at risk to acquire HEV infection than nonpregnant women and HEV infection in this group of Ethiopian pregnant women is associated with high maternal mortality and neonatal complications.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404883 TI - The in vitro activity of Vernonia amygdalina on Leishmania aethiopica. AB - Anti-leishmanial activity of chloroform and methanol extracts of Vernonia amygdalina, a plant widely used in Ethiopia for the treatment of parasitic infections, has been assessed in vitro on Leishmania aethiopica. Amastigotes were more sensitive to V. amygdalina than promastigotes. The chloroform extract had a stronger parasiticidal activity, with median effective doses (ED50) of 18.5 micrograms/ml and 13.3 micrograms/ml for promastigotes and amastigotes, than the methanol extract with ED50 of 74.4 micrograms/ml and 45.8 micrograms/ml respectively. Cytotoxicity caused by V. amygdalina to host cells, the human leukaemia monocyte THP-1 cell line, as determined by the methyl tetrazolium assay, resulted in a median lethal dose (LD50) of 19.6 micrograms/ml for the chloroform extract and 243.4 micrograms/ml for the methanol extract. In comparison, the ED50 and LD50 of pentamidine, a standard anti-leishmanial drug, were 0.5 micrograms/ml and 1.4 micrograms/ml respectively. These results indicate that V. amygdalina displays potent anti-leishmanial activities and warrants further investigation. PMID- 8404884 TI - Longitudinal study of onchocerciasis in Bebeka-Ethiopia. AB - This paper describes the morbidity, intensity and prevalence of onchocerciasis in an endemic area of Ethiopia, using a cross-sectional study (n = 425) and a longitudinal cohort study (n = 98). During the study period (February-October 1987), the increase in intensity and in number of cases with dermal lesions was highly significant (p < 0.005) when compared with the increase in the microfilarial positivity rate (p < 0.1). Since the serial increase in dermatologic problems was not directly correlated with an increase in O. volvulus microfilarial prevalence or intensity, it is suggested that hyper-sensitivity responses to repeated Simulium bites might exacerbate onchocercal dermal lesions. The controversy concerning Simulium bites as a cause of lesions mimicking those due to O. volvulus parasites is discussed. PMID- 8404885 TI - Sero-epidemiological survey of Toxoplasma gondii infection in Ethiopia. AB - In Ethiopia the prevalence of IgG antibodies to Toxoplasma gondii has been determined by an enzyme-linked immunosorbent assay (ELISA). One thousand and sixteen sera collected in six different geographical regions were analyzed. Antibody titres > 15 IU/ml were detected in 74.4% of the specimens, titres exceeding 200 IU/ml in one third of the ELISA-positive sera. The highest antibody titres were found in children and 75% of young adults had sero-converted. As infection with the human immunodeficiency virus 1 (HIV-1) frequently leads to a resurgence of toxoplasmosis, the diagnosis of T. gondii encephalitis should be considered in the clinical management of the HIV-immunocompromised Ethiopian patients. PMID- 8404886 TI - Dracunculiasis (guinea worm disease) in the Bume (Nyangaton) people of South Omo, Ethiopia. AB - A village-to-village search for active dracunculiasis cases was carried out in an endemic area of the Bume (Nyangaton) tribe of South Omo Region, Ethiopia. A total of 21 cases, of which 6, 5, and 10 had pre-emergent, emergent and complicated Guinea worm disease, respectively, were identified. Twenty-two worms, ranging from 1-3 per patient, were removed mainly from the lower limbs; worm appearance seems to be associated more with the right limb. Adults between the ages of 20-30 years are highly affected and infection appears to be sex-related as 14/21 (66.7%) of the cases are females. Water procured from water-holes drug in dry river beds provides an ideal situation for the transmission of dracunculiasis amongst the tribesmen. The knowledge, attitudes and perceptions of the Bume people towards the disease and the public health significance of dracunculiasis are discussed in relation to the current goal of the national and global Guinea worm eradication programme. PMID- 8404887 TI - Intrauterine-neonatal torsion of the spermatic cord in an Ethiopian newborn. AB - The case of a neonate with scrotal swelling noted 6 hours after birth is presented. Scrotal exploration at 12 hours of age revealed a right extravaginal testicular torsion with ischemic necrosis of the testis. Intrauterine and neonatal torsion of the spermatic cord is an uncommon problem accounting for 5 to 6 percent of all cases of testicular torsion. Management controversies revolving around the questions "should the neonate with an obviously infarcted testis be explored?" and "what should be done to the testis whose viability is not reasonably certain?" have now been answered to the satisfaction of most people. This case report from Ethiopia and some aspects of the condition are discussed and attention is called to early diagnosis and appropriate intervention. PMID- 8404888 TI - Peptidylproline cis-trans-isomerases: immunophilins. AB - Two sequence-unrelated families of proteins possess peptidylproline cis-trans isomerase activities (PPIase). PPIases are highly sequence conserved and multifunctional proteins which are present in many types of cells with a considerably divergent phylogenetic distribution. On the cellular level, PPIases occur in every compartment, both as free species and anchored to membranes. Diverse posttranslational modifications such as glycosylation, N-terminal modifications and phosphorylation constitute the additional functional features of PPIases. Folding, assembly and trafficking of proteins in the cellular milieu are regulated by PPIases. These enzymes accelerate the rate of in-vitro protein folding and they have the ability to bind proteins and act as chaperones. Some PPIases are coregulatory subunits of molecular complexes including heat-shock proteins, glucocorticoid receptors and ion channels. Secreted forms of PPIases are inflammatory and chemotactic agents for monocytes, eosinophils and basophils. The potent and clinically useful immunosuppressants CsA, FK506 or rapamycin bind with high affinities to PPIases (immunophilins). The binding criterion allows us to sort the PPIases for the following two superfamilies of proteins: the cyclophilins (CsA-binding proteins) and the FKBP (FK506/rapamycin-binding proteins). Although none of PPIases appeared to be essential for the viability of haploid yeast cells some of the immunophilin/immunosuppressant complexes are toxic both for yeast and mammalian cells. At least seven unlinked genes of cyclophilins and four unlinked genes of FKBP exist in human genomic DNA. Selected immunophilins regulate two different signalling pathways in lymphoid cells, namely the secretion of growth factors by stimulated T-cells and interleukin-2 induced T-cell proliferation. Moreover, selected FKBP mediate the cytotoxic effects of rapamycin in non-lymphoid cells. Accounts of the discovery of PPIases (immunophilins) and their functions are given in this review. A larger spectrum of proteins is analysed in relation to various signal-transduction pathways in lymphoid cells which involve immunophilins or their complexes with the immunosuppressants CsA, FK506 or rapamycin. PMID- 8404889 TI - The L-lactate dehydrogenase gene of the hyperthermophilic bacterium Thermotoga maritima cloned by complementation in Escherichia coli. AB - The gene for a L(+)-lactate dehydrogenase from the hyperthermophilic bacterium Thermotoga maritima was cloned by complementation of an Escherichia coli pfl. Idh mutant. The gene is part of a 4.5 kb SauIIIA fragment obtained by partial digestion of the Thermotoga genome. The DNA fragment was physically mapped and the putative Shine-Dalgarno sequence within the non-coding region determined. The gene contains 960 bp, including the stop codon, corresponding to 319 amino acids/subunit of the homotetrameric enzyme. Part of the amino acid sequence was confirmed by Edman degradation of peptides obtained from nanomolar quantities of the purified enzyme by tryptic digestion. A comparison of the amino acid sequence with those of known prokaryotic L-lactate dehydrogenases reveals a high similarity, especially with the enzyme from thermophilic sources, where up to 48% identity is found. The gene was expressed as an active enzyme in a heterologous host. PMID- 8404890 TI - Rabbit procathepsin E and cathepsin E. Nucleotide sequence of cDNA, hydrolytic specificity for biologically active peptides and gene expression during development. AB - The structure of rabbit procathepsin E was determined by molecular cloning of its cDNA. The proenzyme consisted of 379 amino acids and had structural features common to human and guinea-pig procathepsin E species. The highly conserved tripeptide sequence at the active site of aspartic proteinases, Asp-Thr(Ser)-Gly, is, however, replaced by Asp-Thr-Val in rabbit procathepsin E. To our knowledge, this is the first case of such a variation in aspartic proteinases. The processed form, cathepsin E, hydrolyzed various biologically active peptides maximally at around pH5. Tachykinins, such as substance P and neurokinin A, were hydrolyzed most rapidly, with specific cleavage of sequences essential for their activity. The rates of hydrolysis were several hundred-fold higher than those of cathepsin D. Furthermore, cathepsin E was able to inactivate a functional-domain peptide of fibroblast growth factor, the sequence of which resembles those of tachykinins, and it was active in the generation of functional peptides, such as endothelin and angiotensin I, from their respective precursors. Procathepsin E was detected at high levels in various fetal tissues, such as the liver, stomach and blood cells. At the adult stage, the proenzyme was detectable only in specific tissues, such as the urinary bladder, duodenum and colon. Northern-blot analysis showed similar stage-specific and tissue-specific expression of the mRNA for procathepsin E. Since tachykinins and other suited peptide substrates of cathepsin E have been shown to have mitogenic activity, (pro)cathepsin E may regulate the growth and differentiation of embryonic and fetal tissues by degrading or processing these peptides. The enzyme may also regulate the physiological activities of adult tissues which are mediated by substance P and related tachykinins. PMID- 8404891 TI - Post-translational modifications of the Dictyostelium discoideum glycoprotein PsA. Glycosylphosphatidylinositol membrane anchor and composition of O-linked oligosaccharides. AB - Prespore-specific antigen (PsA) is a cell-surface glycoprotein isolated from Dictyostelium discoideum, which is post-translationally modified by addition of carbohydrate to threonine residues of the carboxy-terminal peptide domain, and a glycosylphosphatidylinositol (GPI) anchor which attaches the glycoprotein to the cell membrane. The GPI anchor was isolated by proteolytic cleavage of the protein, and the structure of the lipid and glycan portions of the anchor were determined. The lipid moiety of the anchor is an inositolphosphoceramide which contains C18:0 phytosphingosine as a long chain base, and a mixture of fatty acids with a C18:1 mono-unsaturated fatty acid as the major component. The purified GPI anchor was susceptible to digestion by a bacterial phosphatidylinositol-specific phospholipase-C enzyme. The glycan of the GPI anchor consisted of two molecular species present in the ratio 55:45, the structures of which were determined by exoglycosidase sequencing and found to be Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 and Man alpha 1-2Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2. The glucosamine in both structures is glycosidically linked to the inositol ring of the inositolphosphoceramide. The GPI glycan structures are consistent with the conserved core structure of all characterised GPI anchors, and the structure of the D. discoideum GPI moiety has features in common with structures from yeast, protozoa and higher eukaryotes. Compositional analysis of the carbohydrate attached to threonine residues in the carboxy-terminal peptide domain is also presented. The oligosaccharides bind to wheat germ agglutinin, and contain glucosamine and fucose as the major constituents. PMID- 8404892 TI - Purification and primary structure of snail metallothionein. Similarity of the N terminal sequence with histones H4 and H2A. AB - A cadmium-binding metallothionein has been purified from metal-exposed Roman snails (Helix pomatia) using gel-permeation, ion-exchange and reverse-phase high performance liquid chromatography. The S-methylated protein was digested with trypsin and the endoproteinases Asp-N, Glu-C and Arg-C. While most of the resulting peptides could be sequenced by Edman degradation, the intact protein, as well as the N-terminal peptide, proved to be blocked. Analysis by mass spectrometry showed that the N-terminal amino acid was an acetylated serine residue. Snail metallothionein, which is suggested to be involved in the detoxification of cadmium, contains 66 amino acid residues, 18 of which are cysteine residues arranged in seven Cys-Xaa-Cys motifs. The calculated molecular mass of the protein is 6.62 kDa. The primary structure of snail metallothionein reveals a clear relationship with molluscan and vertebrate metallothioneins, but lower similarity with metallothioneins of other invertebrate species. The N terminal region of the isolated protein proved to be unique among the metallothionein sequences determined so far, showing high degrees of similarity with the N-terminal sequences of histones H2A and H4 which may be important for regulatory functions. PMID- 8404893 TI - Does oligosaccharide-phosphatidylinositol (glycosyl-phosphatidylinositol) hydrolysis mediate prolactin signal transduction in granulosa cells? AB - Initial biosynthetic radiolabelling experiments with cultured granulosa cells revealed the presence of an oligosaccharide-phosphatidylinositol (glycosyl phosphatidylinositol; (Ose)nPtdIns) structurally related to (Ose)nPtdIns-lipids isolated from other cell types. Prolactin (PRL) stimulated [3H]glucosamine (Ose)nPtdIns turnover and the rapid generation of [3H]myristoyl-diacylglycerol in cultured follicle-stimulating hormone-(FSH)-primed granulosa cells endowed with PRL receptors. In parallel experiments performed with [3H]myo-inositol-labelled granulosa cells, treatment with PRL stimulated (Ose)nPtdIns hydrolysis in a similar manner, whereas no effect on phosphoinositide (PtdIns, PtdInsP and PtdInsP2) turnover could be observed. These results strongly suggest that the cleavage of (Ose)nPtdIns by phosphodiesterase followed by the subsequent generation of diacylglycerol and a soluble phosphoinositol-oligosaccharide (inositol-phosphoglycan; (Ose)nInsP) moiety could be part of the signal transduction mechanism linking PRL receptors to their biological effects in granulosa cells. To test this hypothesis, we examined the effect of PRL and purified (Ose)nInsP moiety (from rat liver membranes) on granulosa cell 3 beta hydroxysteroid dehydrogenase/delta 5-4 isomerase (3 beta-HSD) enzyme activity. Results presented show that, in FSH-primed granulosa cells, PRL (40 nM) and (Ose)nInsP (5 microM) prevented gonadotropin-stimulated 3 beta-HSD activity. Furthermore, in undifferentiated granulosa cells where PRL receptors are absent, no effect of the hormone on 3 beta-HSD activity could be observed, whereas (Ose)nInsP (1-10 microM) inhibited enzyme activity in a dose-dependent manner. PMID- 8404894 TI - A study of colchicine tubulin complex by donor quenching of fluorescence energy transfer. AB - The utility of collisional quenching of energy donors in fluorescence energy transfer is described. In multi-donor single acceptor systems, which contain different classes of donors (as distinguished by their accessibility towards a collisional quencher), donor quenching may be used to assess the fraction of energy transfer from each class of donor. The tubulin-colchicine complex was used as a donor-acceptor system to show that two inaccessible tryptophans are at or near the colchicine binding site. PMID- 8404895 TI - Probes of ligand-induced conformational change in aspartate aminotransferase. AB - Sodium borohydride and sodium cyanoborohydride were assessed as potential reagents for determining ligand-induced changes in accessibility to the active site of aspartate aminotransferase. Rates of reduction of the imine formed between Lys258 and pyridoxal phosphate were determined in the presence of increasing concentrations of the dicarboxylate substrate analogues glutarate and maleate. The rate of reduction decreased to a limiting value which was about 40 fold lower than the equivalent rate in the absence of dicarboxylate. Analysis of the reaction was complicated by the increasing protonation of the imine which accompanied binding of dicarboxylates. Allowing for this increase, the true decrease in accessibility to NaBH3CN was estimated to be approximately 400-fold. Arguments are presented in support of a proposal that the ratio of closed to open conformer of the dicarboxylate-liganded enzyme is approximately 150. The effects of increasing ligand concentration on the reactivity of Cys390 were found to take place in the same range as was observed for NaBH3CN reduction. Conversely, very much higher concentrations of the dicarboxylates were required to protect against proteolysis by trypsin. It is concluded that NaBH3CN reduction and reactivity of cysteine are good determinants of the conformational status of the enzyme but that resistance to tryptic digestion is due to an additional binding mode for the dicarboxylates. PMID- 8404896 TI - Studies on O-glycans of Plasmodium-falciparum-infected human erythrocytes. Evidence for O-GlcNAc and O-GlcNAc-transferase in malaria parasites. AB - O-Glycosylation is the major form of protein glycosylation in human erythrocytes infected with the asexual intraerythrocytic stage of the malaria parasite. Plasmodium falciparum. This study compares aspects of O-glycosylation in P. falciparum-infected and uninfected erythrocytes. Non-labeled and metabolically glucosamine-labeled O-glycans were obtained from the protein fraction of infected or uninfected erythrocytes by beta elimination. Additional label was introduced by reduction with sodium borohydride, or by the attachment of radioactive Gal to peripheral GlcNAc using galactosyltransferase. 2-4-times more labeled O-glycans were obtained from infected erythrocytes compared to the same number of uninfected ones, consistent with additional biosynthesis by the parasite. Our analysis of these O-glycans showed no significant qualitative divergence between the O-glycans of the infected and those of the uninfected red cell. According to preliminary alditol analyses, the O-glycans of P. falciparum-infected red cells do not contain GalNAc at their reducing terminus. Moreover, GalNAc was not synthesized by P. falciparum from either Glc, Gal, GlcN or GalN. At least one O glycan found in P. falciparum-infected erythrocytes contains GlcNAc at its reducing terminus. Gel-filtration results had suggested the presence of O-GlcNAc on proteins in the infected erythrocyte. Probing with a synthetic pentapeptide, we could show that P. falciparum expresses its own O-GlcNAc transferase during intraerythrocytic development. Using this peptide, the enzyme was characterized to some degree. The localization and function of O-GlcNAc in P. falciparum remains to be elucidated. PMID- 8404897 TI - Selective inhibition of N-acetylglucosamine and galactose-specific lectins including the 14-kDa vertebrate lectin by novel synthetic biantennary oligosaccharides. AB - A novel series of synthetic biantennary tri-, penta- and hepta-saccharides with terminal beta-GlcNAc, beta-LacNAc and alpha NeuAc(2,6)beta LacNAc residues, respectively, [LacNAc = Gal beta (1,4)Glc-NAc] connected to a core Gal residue were evaluated for their inhibitory potencies for specific plant and animal lectins. Six isomeric trisaccharides with two beta-GlcNAc residues at the 2,3-, 2,4-, 2,6-, 3,4-, 3,6-, or 4,6-positions of the core Gal were tested for their hemagglutination inhibition activities against two GlcNAc-specific lectins, Griffonia simplicifolia II (GS II) and wheat germ agglutinin (WGA). The 2,3-, 2,4 , 2,6- and 3,6-trisaccharides inhibited WGA 12-50 times more strongly than GlcNAc, whereas only weak or no inhibition was observed with GS II. The 3,4- and 4,6-trisaccharides did not inhibit either of the lectins. Six biantennary isomeric pentasaccharides containing two terminal beta-LacNAc residues with branching patterns similar to the trisaccharides showed selective hemagglutination inhibition of five Gal/GalNAc-specific plant lectins and the 14 kDa Gal-specific calf spleen lectin. The plant lectins include the soybean agglutinin (SBA), ricin agglutinin-I (RCA-I), and three Erythrina lectins with similar specificities: Erythrina indica (EIL), E. corallodendron (ECorL), and E. cristagalli (ECL). The 2,3-pentasaccharide inhibited only SBA and the 14-kDa lectin, and thus was a selective inhibitor among the plant lectins. The 2,6 pentasaccharide inhibited SBA, ECL and the 14-kDa lectin, but not RCA-I or the two other Erythrina lectins. The 4,6-pentasaccharide did not inhibit any of the plant lectins, but was a specific inhibitor of the 14-kDa calf spleen lectin. Synthetic heptasaccharides analogs with 2,4-, 2,6-, 3,6- and 4,6-branching patterns and terminal alpha(2,6)NeuAc residues all showed 25-fold stronger inhibition against the alpha(2,6)sialic-acid-specific elderberry (Sambucus nigra L.) bark lectin as compared to a monovalent disaccharide alpha NeuAc(2,6)beta GalOR. The lack of inhibition of alpha NeuAc(2,6)beta GalOR derivatives methylated at the C6 of the Gal residue and a sulfur-linked thiosialoside derivative demonstrates that the 2,6-anomeric linkage region is important for lectin recognition. Selective inhibition of the Gal/GalNAc-specific lectins was observed for two isomeric C6 methyl-substituted Gal derivatives of methyl beta LacNAc which possess different preferred rotamer orientations about the C5-C6 bond of the Gal residue. PMID- 8404898 TI - The structure of N-linked oligosaccharides of human pancreatic bile-salt dependent lipase. AB - This study describes the structure of the N-linked oligosaccharide chains of bile salt-dependent lipase isolated from the pancreatic juice of a normal donor. After hydrazinolysis, neutral sugar chains were separated from acidic chains by paper electrophoresis and were fractionated using serial column chromatography with immobilized lectins and Bio-Gel P-4 filtration. Structural analysis was performed by means of sequential glycosidase digestion and revealed that the neutral sugar chains are mainly of the biantennary complex type. Fucose residues were identified for some trimannosyl core structures and were alpha(1-6) or alpha(1-2) linked to the innermost GlcNAc residue and a terminal Gal residue, respectively. Sialyl residues were also involved in the oligosaccharide structures. Most of these structures have no linear N-acetyllactosamine repeats. Evidence from several approaches suggests that the sugar chains of the human pancreatic bile salt-dependent lipase possess a blood-group-related antigenic determinant. PMID- 8404899 TI - Variation of the glycosylation of human pancreatic bile-salt-dependent lipase. AB - Glycoproteins of human pancreatic juice were characterized by means of lectins after electrophoresis and electrotransfer to nitrocellulose membranes. For the detected glycoproteins, only a 100-kDa glycoprotein varied in the pancreatic juice from a normal patient (i.e. without any pancreatic disorder) compared to the pancreatic juice from a patient suffering from chronic pancreatitis. This protein, which is the only protein in human pancreatic juice which is O glycosylated and N-glycosylated, was identified as the bile-salt-dependent lipase. Among the glycosylated proteins present in human pancreatic juice, only the glycosylation of bile-salt-dependent lipase differs between individuals. The enzyme was isolated either from normal or pathological human pancreatic juices. The purified variants have an identical molecular mass and amino-acid composition. As suspected from lectin affinity studies, the oligosaccharide composition differs between the variants. The structure of the N-linked oligosaccharides of the variant from the pancreatic juice of a normal donor correlated with complete processing and maturation of a complex-type N-glycan. Alteration of the maturation process can be detected for a bile-salt-dependent lipase variant from a patient suffering with chronic pancreatitis, since the carbohydrate composition is compatible with the predominance of hybrid or high mannose-type structures. The amount of sugar involved in O-glycosylation associated with the peanut agglutinin reactivity suggests the presence of 12-14 minimal Gal beta 1-->3GalNac-->T/S O-glycan structures which are sialylated and fucosylated. The amount of sugar involved in the O-linked oligosaccharide structure appears to be unchanged in the variants isolated from the pathological pancreatic juice. PMID- 8404900 TI - GTPase activity of small GTP-binding proteins in HL-60 membranes is stimulated by arachidonic acid. AB - The GTPase activity of membranes isolated from differentiated HL-60 cells was investigated to obtain information about the possible involvement of membrane bound GTP-binding proteins in the regulation of the NADPH oxidase. A more than tenfold increase in the rate of hydrolysis of membrane-bound GTP was observed when cytosol and arachidonic acid were added simultaneously, i.e. under the same conditions where NADPH oxidase becomes activated. There were parallel changes in GTPase and NADPH oxidase activities when the concentration of arachidonic acid or the species of the fatty acid was varied or different detergents were applied. Separation of the GTP-binding proteins of the solubilized membrane by sucrose density gradient centrifugation, allowed us to ascribe the observed effect to the stimulation of the GTPase activity of small GTP-binding proteins by cytosolic component(s). Indirect evidence suggests that, in contrast to the effect upon recombinant ras and ras-GTPase-activating protein, in intact HL-60 membranes the interaction of rap1A with rap-GTPase-activating protein, is strongly enhanced by arachidonic acid. PMID- 8404901 TI - Erythropoietin induces the association of phosphatidylinositol 3'-kinase with a tyrosine-phosphorylated protein complex containing the erythropoietin receptor. AB - Stimulation of sensitive cells with erythropoietin results in rapid induction of protein tyrosine phosphorylation. Other than tyrosine phosphorylation of one chain of the erythropoietin receptor, the identities of the remaining tyrosine phosphorylated proteins are undefined. In this report, we demonstrate that the stimulation of the erythropoietin-sensitive human UT7 cells by erythropoietin rapidly resulted in the appearance of phosphatidylinositol 3-kinase activity in anti-phosphotyrosine immunoprecipitates. Erythropoietin action was rapid, detectable after as early as 1 min stimulation, transient, returning to control level after 30 min stimulation and was observed using the erythropoietin concentrations able to stimulate the cell proliferation. Anti (phosphatidylinositol 3-kinase) antibodies specifically immunoprecipitated 125I erythropoietin bound to its receptor, strongly suggesting that phosphatidylinositol 3-kinase associated with a protein complex containing the activated erythropoietin receptor. To confirm this result, phosphatidylinositol 3 kinase was immunoprecipitated from erythropoietin-stimulated cells using mild conditions followed by Western analysis using anti-phosphotyrosine antibodies. Five tyrosine phosphorylated proteins were revealed: the cloned chain of the erythropoietin receptor, the regulatory subunit of phosphatidylinositol 3-kinase and three unidentified proteins of 111, 97 and 64 kDa. None of these tyrosine phosphorylated proteins was detected in anti-(phosphatidylinositol 3-kinase) immunoprecipitates from unstimulated cells. Thus, our results show that phosphatidylinositol 3-kinase associates with a tyrosine-phosphorylated protein complex containing the activated erythropoietin receptor. PMID- 8404902 TI - Determinants for the enhanced thermostability of hybrid (1-3,1-4)-beta glucanases. AB - Hybrid (1-3,1-4)-beta-glucanases which contain an N-terminal region derived from the Bacillus amyloliquefaciens enzyme and a C-terminal region of the closely related B. macerans enzyme may exhibit a thermostability superior to both parental enzymes. A systematic series of hybrid enzymes were constructed in order to delineate the amino acid residues that affect protein stability. Hybrid enzymes with between one and four of the N-terminal residues for the mature B. amyloliquefaciens (1-3,1-4)-beta-glucanase exhibit no significant changes in biochemical characteristics as compared with the parental B. macerans enzyme. However, significantly enhanced thermostability was observed in the hybrid enzyme containing an N-terminal segment of eight amino acid residues derived from the B. amyloliquefaciens enzyme. Site-directed mutagenesis revealed that the combined effect of Gln1, Thr2, Ser5 and Phe7 confer enhanced stability on hybrid enzymes, probably by improving the hydrogen bonding that stabilizes the interactions between the N-terminal and the centre of the folded molecule, as well as between the two termini of the polypeptide chain. Furthermore, deletion of Tyr13 in the hybrid enzyme containing the 12 N-terminal amino acids from the B. amyloliquefaciens (1-3,1-4)-beta-glucanase results in a dramatic increase in stability at 70 degrees C with the half-life of 6 min increased to around 4 h. This is twofold higher than the hitherto most stable hybrid enzyme in which the N terminal domain consisted of 16 residues of the B. amyloliquefaciens enzyme. PMID- 8404903 TI - Biosynthesis of platelet-activating factor in cultured mast cells. Involvement of the CoA-independent transacylase demonstrated by analysis of the molecular species of platelet-activating factor. AB - We have recently demonstrated that arachidonate [20:4(5,8,11,14)] was primarily linked to the hexadecyl (16:0) and octadecenyl (18:1) species of alkylacyl derivatives of glycerolphosphocholine (GroPCho). Consistent with the involvement of arachidonate-specific CoA-independent transacylase in the synthesis of platelet-activating factor (PAF; 1-O-alkyl-2-acetyl-GroPCho), 16:0 and 18:1 PAF species were formed upon antigen stimulation [Joly, F., Breton, M., Wolf, C., Ninio, E. & Colard, O. (1992) Biochim. Biophys. Acta 1125, 305-312]. In the present work, addition of lyso-PAF to mast cells resulted in PAF production. We analyzed the PAF species formed in the presence of a defined lyso-PAF molecular species in order to differentiate between either direct acetylation or involvement of the membrane precursor. The 18:1 lyso-PAF was more effective than the 16:0 in producing PAF which was composed of 95% 18:1 PAF, the balance being 16:0, indicating that part of the acetylated lyso-PAF originated from the cellular pool of alkyl-arachidonyl-GroPCho in resting cells. Consistent with alkyl-arachidonyl-GroPCho species content and acetyltransferase specificity, similar amounts of 16:0 and 18:1 PAF species were formed when mast cells were stimulated with antigen. Supplemented with 16:0 or 18:1 lyso-PAF, antigen stimulated mast cells responded by 230% and 125% increase in PAF synthesis, respectively. As expected, the amount of the PAF species corresponding to the added lyso-PAF was increased. More interestingly, addition of 16:0 lyso-PAF almost doubled the amount of 18:1 PAF content as compared to antigen alone, thus indicating that the lyso-PAF formed via the CoA-independent transacylase was significantly used for PAF synthesis, despite a large excess of exogenous lyso PAF. The CoA-independent transacylase, measured using [3H]lyso-PAF as a substrate in sonicates from antigen-stimulated cells, was decreased concurrently with PAF formation. In conclusion, we show that when lyso-PAF is added to mast cells, a direct acetylation may occur. However, PAF is preferentially synthesized through a mechanism involving the CoA-independent transacylase reaction. PMID- 8404904 TI - Purification of trehalose synthase from baker's yeast. Its temperature-dependent activation by fructose 6-phosphate and inhibition by phosphate. AB - A trehalose synthase purified from baker's yeast contained 56-kDa, 102-kDa and 123-kDa polypeptides as its main components. The 102-kDa polypeptide was isolated and shown to be a specific trehalose-6-phosphatase. The trehalose-6-phosphate synthase (Tre6P synthase) activator described by Londesborough and Vuorio [(1991) J. Gen. Microbiol. 137, 323-330] was shown to be phosphoglucoisomerase and to function entirely by generating fructose 6-phosphate. Below 35 degrees C, fructose 6-phosphate is a powerful activator of the Tre6P synthase activity of intact trehalose synthase, especially at physiological phosphate concentration, but does not affect its trehalose-6-phosphatase activity nor the Tre6P synthase activity of truncated trehalose synthase containing truncated versions of the 123 kDa polypeptide. At 50 degrees C, activation by fructose 6-phosphate and inhibition by phosphate are greatly decreased, resulting in an unusually high temperature-dependence for the Tre6P synthase activity at a physiological phosphate concentration (2 mM). PMID- 8404905 TI - Cloning of two related genes encoding the 56-kDa and 123-kDa subunits of trehalose synthase from the yeast Saccharomyces cerevisiae. AB - Preparations of intact trehalose synthase contain three polypeptides with molecular masses of 56, 102 and 123 kDa. We have cloned the genes TSS1 and TSL1 coding for the 56- and 123-kDa subunits, respectively. These genes are located on chromosomes II (TSS1) and XIII (TSL1). The TSS1 gene was found to be identical with CIF1, a gene required for normal growth on glucose. The product of the entire TSS1 gene exhibits 37% identity with a 502-amino-acid stretch from the middle of the TSL1 product. Disruption of the TSS1 gene in yeast eliminates both trehalose 6-phosphate synthase (Tre6P synthase) and trehalose 6-phosphate phosphatase (Tre6Pase) activities, and reintroduction of this gene restores these activities. Transformation of Escherichia coli with TSS1 increases its Tre6P synthase activity. Specific proteolytic degradation of the 123-kDa polypeptide from the N-terminus greatly influences the Tre6P synthase activity, decreasing its inhibition by phosphate and activatability by fructose 6-phosphate but has little effect on the Tre6Pase activity. These results suggest that this N terminal part confers regulatory properties upon the Tre6P synthase activity. PMID- 8404906 TI - The amino acid sequence of Ole e I, the major allergen from olive tree (Olea europaea) pollen. AB - The complete primary structure of the major allergen from Olea europaea (olive tree) pollen, Ole e I (IUIS nomenclature), has been determined. The amino acid sequence was established by automated Edman degradation of the reduced and alkylated molecule as well as of selected fragments obtained by proteolytic digestions. Ole e I contains a single polypeptide chain of 145 amino acid residues with a calculated molecular mass of 16331 Da. No free sulfhydryl groups have been detected in the native protein. The molecule contains a putative glycosylation site. A high degree of microheterogeneity has been observed, mainly centered in the first 33% of the molecule. Comparison of Ole e I sequence with protein sequence databases showed no similarity with other known allergens. However, it has a 36% and 38% sequence identity with the putative polypeptide structures, deduced, respectively, from nucleotide sequences of genes isolated from tomato anthers and corn pollen, which have been suggested to be involved in the growing of the pollen tube. Therefore, the olive tree allergen may be a constitutive protein of the pollen involved in reproductive functions. PMID- 8404907 TI - Application of an ectopic expression system for the selection of protein-isoform specific antibodies. The monoclonal antibody K1C3 is specific for the RCK1 potassium channel. AB - Monoclonal antibodies were raised against a fusion protein consisting of a fragment of 141 amino acids of the C-terminal region of the rat brain voltage gated K(+)-channel protein (RCK1) and the lambda N protein (fusion protein I). Selection of K(+)-channel-specific hybridoma cell lines was performed by means of an ELISA employing a fusion protein consisting of the K(+)-channel-specific peptide sequence and glutathione S-transferase (fusion protein II). For final selection of RCK1 isoform-specific antibodies, a panel of Xenopus oocytes was employed, each injected with cRNA coding for a specific RCK isoform (RCK 1, 2, 4 or 5). Several days after injection, cryosections of embedded oocytes were obtained and were employed in immunohistochemical analysis of antibody binding. Of five hybridoma supernatants from stable growing hybridoma cell lines, selected by the fusion-protein ELISA, one monoclonal antibody (denoted K1C3) recognized exclusively the RCK1-protein isoform, with the other four exhibiting different levels of cross-reactivity with other K(+)-channel isoforms, or with unknown protein(s) of non-injected oocytes. The expression of the RCK1 protein in the postnatal brain was studied using, as far as we are aware, the first example of the application of such isoform-specific antibodies. PMID- 8404908 TI - Chemical structures of the core regions of Campylobacter jejuni serotypes O:1, O:4, O:23, and O:36 lipopolysaccharides. PMID- 8404909 TI - Controversies in the laboratory diagnosis of community-acquired urinary tract infection. AB - Urine samples constitute the largest single category of specimens examined in most medical microbiology laboratories. The everyday nature and apparent simplicity of urinary tract infection belies the intense debate and controversy regarding the optimal methods of collection, transport and processing of urine specimens and reporting of results. There is considerable variation in the interpretation of quantitative culture results between laboratories and the etiology of abacterial cystitis remains unclear. Microscopy to detect pyuria provides information on an important indicator of inflammation and it has been proposed that detection of urinary antibody may provide similar information. Neither of these indices of host response is suitable for use in a screening test for urinary infection however, although they may usefully contribute to the interpretation of significance of culture results. The development of screening tests and automated systems continues, but at present microscopy and culture remain the most important techniques for laboratory diagnosis. However, these techniques have so far failed to provide an etiological diagnosis for abacterial cystitis and this remains a major area for research. PMID- 8404910 TI - Re-emergence of meningococcal carriage on three-year follow-up of a kibbutz population after whole-community chemoprophylaxis. AB - A long-term study was conducted to determine the rate of re-emergence of throat carriage of meningococci in a semi-closed kibbutz community after the administration of chemoprophylaxis to all its members. Serotype B:4 was selected as marker organism since it was isolated from a fatal case and was the most frequently occurring strain (80%) among serogroup B isolates, which themselves comprised 54% of all meningococci. The carriage rate among Israeli residents (volunteer workers were analyzed separately) before treatment was 6.6% (49/748) overall, with 4.3% group B strains. Three weeks after treatment, in most cases with rifampicin (whereby three persistently positive persons were retreated with minocycline), no meningococci were recovered. Six months later, 1.9% of a population sample aged < or = 30 years were positive, while before treatment and one and three years later, 9.4%, 8.6% and 4.6% respectively were positive in this age group. Serotype B:4 comprised 81.3% of group B strains before prophylaxis, 5.3% after one year, and 28.6% after three years, thus possibly re-establishing itself as the single dominant serotype. The marked suppression of carriage after mass chemoprophylaxis appeared to last at least six months, with the meningococcal population being re-established within a year. PMID- 8404911 TI - Evaluation of agents for use in medium for selective isolation of Lyme disease and relapsing fever Borrelia species. AB - Barbour-Stoenner-Kelly (BSK) II medium containing fosfomycin, 5-fluorouracil, trimethoprim and sulfamethoxazole was evaluated for the selective isolation of the Borrelia species responsible for Lyme disease and relapsing fever. The maximum non-inhibitory concentrations of fosfomycin, 5-fluorouracil, trimethoprim and sulfamethoxazole for six strains of borreliae were 500 to > 1000 micrograms/ml, 250 to > 500 micrograms/ml, 125 to 500 micrograms/ml and 125 to 500 micrograms/ml, respectively. The combination of four agents (fosfomycin 400 micrograms/ml, 5-fluorouracil 100 micrograms/ml, trimethoprim 10 micrograms/ml, sulfamethoxazole 50 micrograms/ml) did not inhibit the growth of borreliae, allowing growth in cultures inoculated with a few organisms (theoretically a single organism). In contrast, the four-agent combination completely inhibited the growth of 12 of 13 other bacterial strains tested as possible contaminants. This combination also allowed the selective growth of borreliae in experimentally contaminated specimens. The four-agent combination in BSK II medium may be useful for selective isolation of Borrelia species responsible for Lyme disease and relapsing fever from clinical and environmental samples. PMID- 8404912 TI - Effect of pathological changes of pH, pO2 and pCO2 on the activity of antimicrobial agents in vitro. AB - Since standard susceptibility tests reflect the physiological rather than the pathological conditions prevailing within an infected abdomen, as recently documented, the effect of reduced pH and pO2 and increased pCO2 on the activity of antibiotics in vitro was studied. MICs were determined in vitro under standard culture conditions (MICstandard) and modified conditions (MICmodification) simulating the previously determined pathological values. Various classes of antibiotics were affected differently by the modified conditions. However, within an antibiotic class similar results were obtained for gram-negative and gram positive pathogens. Median MICmodification/MICstandard ratios were 4 for aminoglycosides, 2 for quinolones and clindamycin, 1 for cephalosporins, and 0.5 for penicillins and vancomycin. Anaerobic conditions and a pH of 6.4 further increased the ratio of aminoglycosides to 8. Ratios were similar within an antibiotic class at inocula of 10(5) or 10(7) cfu/ml. All MICs determined in tests with imipenem against gram-negative and gram-positive bacteria and with vancomycin against gram-positive organisms were below the susceptibility breakpoint, whatever conditions and inocula were employed. In contrast, the percentage of MICs in susceptibility range using high inocula and modified conditions decreased to 78% for penicillins, 73% for cephalosporins, 22% for aminoglycosides, 11% for quinolones and 0% for clindamycin. In conclusion, routine susceptibility testing may overestimate the activity of aminoglycosides and underestimate the activity of beta-lactams under the conditions prevailing during abdominal infection. PMID- 8404913 TI - Use of a DNA hybridization method to verify results of screening for methicillin resistance in staphylococci. AB - Tests were performed by the disk diffusion method, agar dilution method and the E test to determine the susceptibility to methicillin and oxacillin of clinical isolates and control strains of Staphylococcus aureus (n = 106) and coagulase negative species (n = 131). Results were compared with those of a dot blot DNA hybridization test, in which the mecA gene was detected using an oligonucleotide probe selected from the mecA gene. Among the Staphylococcus aureus strains the mecA gene was found in all but two strains inhibited by > or = 8 mg/l of methicillin and all but two strains inhibited by > or = 4 mg/l of oxacillin. A disk test using either 1 microgram oxacillin or 10 micrograms methicillin and a tentative resistance breakpoint of < or = 10 mm gave the best agreement with the hybridization test. For coagulase-negative staphylococci 34 of 35 strains inhibited by > or = 8 mg/l methicillin hybridized with the probe as well as 58 of 82 strains inhibited by 1-4 mg/l; 93 of 97 strains inhibited by > or = 0.5 mg/l oxacillin were also positive in the probe test. Using the 1 microgram oxacillin disk and a resistance breakpoint of < or = 10 mm good agreement was obtained between results of the disk diffusion and DNA hybridization tests. It is suggested that this genotypic method for detection of methicillin resistance is used as a reference method for routine methods. PMID- 8404914 TI - Effect of alkalinisation and increased fluid intake on bacterial phagocytosis and killing in urine. AB - The effect of alkalinisation and increased fluid intake on bacterial phagocytosis and killing in urine was studied. Phagocytosis of Escherichia coli and Staphylococcus saprophyticus by polymorphonuclear neutrophils (PMN) took place in only one of three first voided early morning urine samples from volunteers, and no bacterial killing occurred. This was attributed to the high osmolality (690 to 720 mOsm) and low pH of the early morning urine; two samples were pH 5.8 and the third, in which phagocytosis occurred, was pH 6.4. Afternoon urine samples from the same volunteers had lower osmolality and a higher pH, with a high rate of phagocytosis (> or = 88%) and 55-69% killing. PMN remained viable (mean 94%) after exposure for 1 h to both early morning and afternoon urine. Volunteers taking 4 g sodium citrate showed a mean rise of urinary pH of 1 unit. Water loading in patients with urinary tract infections produced a bacteria to neutrophil ratio conducive to bacterial killing, bacterial counts being reduced by a mean of 2.5 logs after 1-3 h without a corresponding reduction in neutrophils. Thus, raising the pH and reducing the osmolality of urine increases the ability of neutrophils to eliminate infecting organisms. PMID- 8404915 TI - Asymptomatic Neisseria subflava biovar perflava bacteriuria in a child with obstructive uropathy. AB - A case of asymptomatic urinary tract infection with Neisseria subflava biovar perflava in a 10-year-old male patient with congenital structural abnormalities of the urinary bladder is presented. The organism was recovered from three catheter urine specimens collected over a seven-month period. A brief review of the role of saprophytic Neisseria species in infectious processes is presented and the likely source of this unusual urinary tract isolate is discussed. PMID- 8404916 TI - Diagnostic value of determination of serum mannan concentrations in patients with candiduria. AB - The diagnostic value of determination of serum mannan levels was studied in 41 patients with candiduria and 20 control patients without candiduria. The patients with candiduria comprised 18 asymptomatic afebrile patients (group 1) and 23 febrile patients with suspected Candida pyelonephritis (group 2). Mannan antigenemia (> or equal to 0.5 ng/ml) was detected in 14 patients in group 2, this rate being significantly higher than the rate of four patients in group 1 (p < 0.05) and none in the control group (p < 0.01). Serum mannan levels decreased gradually and became undetectable in six patients in whom therapy with an anticandidal agent was effective. Determination of serum mannan levels may be a useful parameter in diagnosing Candida pyelonephritis and deciding on effective treatment. PMID- 8404917 TI - In vitro activity of vancomycin and teicoplanin against Staphylococcus aureus and Staphylococcus epidermidis colonizing catheters. AB - In a quantitative in vitro model the activity of vancomycin and teicoplanin in two concentrations (4 x MBC and 1 mg/l) against Staphylococcus aureus and a slime producing Staphylococcus epidermidis strain colonizing the internal surface of polyurethane and silicone catheters was studied. In comparison with vancomycin, teicoplanin achieved a significantly greater reduction (p < 0.05) in the counts of Staphylococcus aureus and Staphylococcus epidermidis adhering to both polyurethane and silicone catheters. PMID- 8404918 TI - Haemophilus test medium versus Mueller-Hinton broth with lysed horse blood for antimicrobial susceptibility testing of four bacterial species. AB - Studies were undertaken to determine whether broth microdilution susceptibility tests could be standardized by using a single medium for testing fastidious respiratory pathogens. Mueller-Hinton broth with lysed horse blood and the broth version of Haemophilus Test Medium (HTM) were directly compared. Ten orally administered agents were found to give essentially identical results in both media but minor differences were noted. Because the test are easier to read when HTM broth is used, that medium is to be preferred for routine testing of Haemophilus influenzae, Streptococcus pneumoniae, Streptococcus pyogenes and Moraxella catarrhalis isolates by the microdilution procedure. PMID- 8404919 TI - Value of measurement of neutrophil elastase-alpha 1 proteinase inhibitor levels in the early diagnosis of neonatal infection. AB - In a case-control study in 398 neonates the value of measuring the levels of neutrophil elastase-alpha 1 proteinase inhibitor (EPI) for early diagnosis of neonatal infection was studied in comparison to the commonly used parameters of leukocyte count, ratio of immature to total granulocytes and C-reactive protein levels. Investigations were performed on day 1 or day 6 of life. On the basis of the clinical findings patients were allocated to one of the three following groups: healthy neonates (group A), neonates with local infections such as pneumonia or skin infection (group B) or neonates with septicemia as demonstrated by a positive blood culture (group C). The median EPI levels (with range) measured on day 1 were: group A 40 (15-65) micrograms/l, group B 120 (80-260) micrograms/l, group C225 (140-355) micrograms/l. The levels on day 6 were: group A 27.5 (5-55) micrograms/l, group B 105 (65-370) micrograms/l, group C 182.5 (74 450) micrograms/l. EPI thus discriminated well between healthy neonates and neonates with infection, but not between neonates with infection and neonates with septicemia. PMID- 8404920 TI - Duration of specific immunoglobulin A antibody following acute toxoplasmosis as determined by enzyme immunoassay and immunosorbent agglutination assay. AB - Modifications of an enzyme immunoassay (EIA) and an immunosorbent agglutination assay (ISAGA) for measuring Toxoplasma gondii-specific IgM antibody were made to enable the measurement of Toxoplasma gondii-specific IgA antibody. It was shown that specific IgA could be measured by both assays but that the ISAGA was slightly more sensitive. IgA appears about two weeks after IgM and persists for 6 to 7 months. However, the IgA response varies considerably both in degree and duration, and demonstration of IgM antibody is at present the most suitable routine test for the diagnosis of recent Toxoplasma gondii infection. PMID- 8404921 TI - In vitro susceptibility of Helicobacter pylori to trospectomycin, pirlimycin (U 57930E), mirincamycin (U-24729A) and N-demethyl clindamycin (U-26767A). AB - The in vitro activity of trospectomycin, pirlimycin, mirincamycin and N-demethyl clindamycin was measured against 46 clinical isolates of Helicobacter pylori using an agar dilution technique. The MIC50 and MIC90 were 4 and 64 micrograms/ml for pirlimycin and N-demethyl clindamycin, and 32 and 128 micrograms/ml for mirincamycin, respectively. All 46 strains were sensitive to trospectomycin with an MIC50 of 8 micrograms/ml and an MIC90 of 16 micrograms/ml. Of seven strains with the highest trospectomycin MICs (8 or 16 micrograms/ml) 100% were found to be resistant to metronidazole. Among ten strains with low trospectomycin MICs (2 micrograms/ml or less) 100% were sensitive to metronidazole. Possible explantations for the apparent correlation between the MICs of the two drugs are discussed. Since all metronidazole resistant strains were sensitive to trospectomycin, this drug may be useful in treating infection with metronidazole resistant Helicobacter pylori. PMID- 8404922 TI - Clarithromycin-induced leukocytoclastic vasculitis. PMID- 8404923 TI - Nasal Mycobacterium kansasii infection in a patient with acquired immunodeficiency syndrome. PMID- 8404924 TI - Isolation of Campylobacter jejuni strains resistant to nalidixic acid and fluoroquinolones from children with diarrhea in Athens, Greece. PMID- 8404925 TI - Isolation of an unclassified non-fermentative gram-negative rod from a patient on continuous ambulatory peritoneal dialysis. PMID- 8404926 TI - Primary cytomegalovirus infection associated with the onset of ulcerative colitis. PMID- 8404927 TI - Prevalence of antibodies to hepatitis A virus in health care workers. PMID- 8404928 TI - Breakpoint determination: rufloxacin. PMID- 8404929 TI - Breakpoint determination: piperacillin/tazobactam. PMID- 8404930 TI - Dobutamine stress--Doppler echocardiography before and after coronary angioplasty. AB - To determine if dobutamine-induced myocardial ischaemia causes abnormalities in Doppler parameters of left ventricular ejection and filling and to assess early effects of successful coronary angioplasty (PTCA) on these parameters, dobutamine stress echocardiography and Doppler studies were performed once in 11 normal volunteers and twice in 17 patients (within 1 day pre- and post-PTCA). Dobutamine induced wall motion abnormalities, ST changes and angina in 11, five and five patients, respectively, before and three, two and one patients, respectively, after PTCA. Doppler indices of both systolic and diastolic function were comparable at rest, before and after PTCA. Dobutamine induced similar increases in peak aortic velocity and average acceleration in healthy individuals (39% and 53%) and in patients with one-vessel disease both before (38% and 39%) and after PTCA (39% and 40%). In the three patients with multivessel disease, peak aortic velocity showed a blunted response (-0.3%) before PTCA but increased by 17% after PTCA, while acceleration decreased both before (12%) and after PTCA (14%). There were significant differences (P < 0.0001) between healthy individuals and pre PTCA patients in the effect of dobutamine on peak early (E) filling velocity (+34% vs -19%), E-acceleration (+35% vs -26%), peak early to atrial filling velocity ratio (E/A) (-0.7% vs -37%) and diastolic time velocity integral (TVI) (+34% vs -22%). After PTCA, the response of Doppler diastolic indices improved during dobutamine, as shown by the increase in E and E-acceleration (+8%, +24%), respectively) and by the decline in the reduction of E/A and TVI (-17% and -10%, respectively). Thus, the response of Doppler diastolic parameters to dobutamine stress is a sensitive indicator of significant coronary disease and is superior to changes in ejection indices. Successful PTCA resulted in an improved diastolic filling response to dobutamine stress. PMID- 8404931 TI - The relationship between early plasma atrial natriuretic factor levels and exercise performance after myocardial infarction. AB - We tested the hypothesis that early plasma atrial natriuretic factor (ANF) values are related to subsequent functional capacity in patients with acute myocardial infarction (MI). Blood for ANF determination was sampled from 90 male patients (age 66.5 +/- 9.5 (mean +/- SD) years) day 3 post MI. Exercise testing on an upright bicycle ergometer to symptomatic end-points was performed 1 and 6 months after MI in 83 and 78 patients, respectively. A weak, but significant inverse relationship between day 3 plasma ANF levels and exercise duration after MI (1 month: r = -0.27, P = 0.012; 6 months: r = -0.36, P = 0.001) was observed. In the subgroup of patients without effort-associated ischaemia, the relationship was closer (1 month; n = 38, r = -0.57, P < 0.001; t months: n = 33, r = -0.65, P < 0.001). In multivariate analysis, with ANF, patient age and peak creatine kinase MB values as covariates, the relationship remained significant. These findings suggest that in male patients subacute plasma ANF measurements are predictive of exercise capacity following acute MI. The relationship appears to be especially prominent in patients without effort-related ischaemia during exercise. PMID- 8404932 TI - Lipid peroxidation and antioxidant status following thrombolytic therapy for acute myocardial infarction. AB - We have investigated the timescale of increased lipid peroxidation following successful early thrombolytic therapy for acute myocardial infarction and report for the first time reciprocal changes in plasma chain-breaking antioxidants. Sixty-seven patients were recruited following a first acute myocardial infarction within 6 h of the onset of symptoms and received 70 or 100 mg of recombinant tissue plasminogen activator (Actilyse) as two intravenous bolus injections 30 min apart. Serial blood samples were taken before administration of thrombolytic therapy and after 30 min, 60 min, 90 min, 6 h and 24 h. Coronary artery patency was assessed at 90 min by coronary angiography. Malondialdehyde (MDA), a marker of lipid peroxidation, and the chain-breaking antioxidants alpha-tocopherol, retinol and ascorbate were measured by high performance liquid chromatography. When the coronary artery was patent there was an early rise in plasma MDA (time 0 0.91 +/- 0.05 mumol.l-1) with levels peaking at 90 min (1.02 +/- 0.06, P < 0.05) and returning to baseline by 6 h (0.85 +/- 0.06), accompanied by reciprocal decreases in alpha-tocopherol (time 0 7.13 +/- 0.34 mumol.mmol-1 cholesterol, 90 min 6.64 +/- 0.33, P < 0.05) and retinol (time 0 1.99 +/- 0.10 mumol.l-1, 90 min 1.81 +/- 0.08, P < 0.05). Ascorbate levels did not change significantly until 24 h (time 0 29.5 +/- 4.9 mumol.l-1, 24 h 22.6 +/- 4.4, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404933 TI - Prognostic significance of the evolution of left ventricular ejection fraction in patients with acute myocardial infarction not treated with thrombolytic therapy. AB - Several controlled trials on the thrombolytic treatment of acute myocardial infarction (AMI) have failed to demonstrate that thrombolysis has a simultaneous positive effect on left ventricular function and survival. One explanation may be that spontaneous changes in left ventricular function occurred during the progression of AMI in control patients. The aim of this study was to evaluate the spontaneous evolution of left ventricular ejection fraction (LVEF) and its prognostic influence on early (1 month) and late (1 year) mortality in patients with AMI. We studied 216 patients admitted to our CCU within 24 h of the onset of symptoms. LVEF was determined by radionuclide ventriculography on admission (RNV1) and at the end of the necrotic phase (RNV2). Fourteen patients died before RNV2. On the basis of LVEF values at RNV1, the remaining 202 patients were divided into two groups: those with a normal LVEF (> or = 55%), and those with an abnormal LVEF (< 55%). Among patients with a normal LVEF at RNV1 (64 patients), a significant increase (> 12%) in LVEF at RNV2 was observed in 12.5%, a significant decrease (> 12%) in 12.5% and no change at all in 75%. All of these patients survived, regardless of the evolution of LVEF. In patients with an abnormal LVEF at RNV1 (138) a significant increase (> 5%) in LVEF at RNV2 was observed in 72.5%, a significant decrease (> 5%) in 6.5% and no change at all in 21%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404934 TI - Adverse effect of warfarin in acute myocardial infarction: increased left ventricular thrombus formation in patients not treated with high-dose heparin. AB - In a prospective non-randomized study, 229 patients with a verified first acute anterior myocardial infarction (AAMI) underwent echocardiography before discharge in order to study left ventricular (LV) thrombus formation. Antithrombotic therapy was given according to the routine of each centre. Patients receiving high-dose heparin had few LV thrombi, irrespective of warfarin therapy (6/32 vs 3/25, P ns). In patients not given heparin, however, a significantly higher prevalence of LV thrombi was found in a subgroup of patients treated with warfarin as compared to those who did not receive warfarin (8/13 vs 17/68, P 0.02). A similar, but non-significant difference was observed in patients given low-dose heparin (42% vs 27%, P ns). Within the non-heparin and low-dose heparin groups, age, infarct size, occurrence of Q-wave infarction, congestive heart failure and LV wall motion impairment did not differ between those treated or not treated with warfarin. In conclusion, high-dose heparin seems effective in the prevention of LV thrombosis irrespective of warfarin therapy after AAMI. The start of warfarin therapy in patients not receiving heparin was, however, associated with an increased prevalence of LV thrombosis. PMID- 8404935 TI - Late thrombolysis followed by antithrombotic treatment in acute myocardial infarction: effect of therapy evaluated with thallium-201 SPECT. AB - Perfusion abnormalities and infarct size evaluated by thallium-201 SPECT after late thrombolysis followed by anti-thrombotic treatment in acute myocardial infarction were studied in 111 patients. Patients were prospectively randomized to a treatment group (n = 54) receiving streptokinase initiated 12-49 h after onset of symptoms, followed by heparin for at least 5 days and warfarin and dipyridamole for at least 3 months, or to a control group (n = 57) treated routinely without any of these drugs. Thallium-201 scintigraphy was performed within 3 months of follow-up in 40 (74%) of the patients in the treatment group and 46 (81%) in the control group. During a symptom-limited bicycle test the isotope was injected at peak exercise. SPECT was acquired 10 min later (exercise SPECT) and after 3 h (redistribution SPECT). A computerized quantitative analysis was performed. Defects were defined as regions with activity below 40% of the maximum. In the exercise SPECT defects were significantly smaller in the treatment group than among control patients (9 +/- 12% vs 15 +/- 15% of the left ventricle; P < 0.05). The difference was more apparent in patients with Q-wave than in those with non-Q-wave infarctions and the entire difference was accounted for by patients without a previous history of infarct or angina pectoris. There was no statistically significant difference in defect size in the redistribution SPECT (5 +/- 10% vs 9 +/- 11%, ns).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404936 TI - Balloon mitral valvotomy with a single catheter: a comparison between bifoil/trefoil and the Inoue balloon. AB - Results of percutaneous mitral valvotomy were compared in two groups undergoing the procedure at our institution. Group I: 100 patients having had percutaneous valvotomy with the Schneider-Medintag bifoil (2 x 19 mm) or trefoil (3 x 15 mm or 3 x 12 mm) catheters, and group II; 150 patients in whom the procedure was performed with the Inoue balloon (24-30 mm). Baseline clinical (age, gender, NYHA class and echo score) and haemodynamic variables were similar in both groups. Haemodynamic improvement occurred in both groups. Although the reduction in left atrial pressure did not differ significantly between the two groups, the increase in mitral valve area was significantly (P < 0.001) higher for group I (0.8 +/- 0.2 to 1.9 +/- 0.7 and 0.8 +/- 0.3 to 1.6 +/- 0.3 cm2 respectively for mitral area, and 22 +/- 6 to 13 +/- 5 and 21 +/- 6 to 13 +/- 5 mmHg respectively for mean left atrial pressure). The increase in cardiac output was statistically significant in group I (3.2 +/- 0.7 to 4.0 +/- 0.9 l.min-1, P < 0.05) but not in group II (3.5 +/- 2.0 to 3.7 +/- 0.9 l.min-1, ns). Inter-atrial shunting immediately after valvotomy was recorded in 19% of group I patients compared with 6% in group II (P < 0.001). The overall incidence of significant mitral regurgitation (3+ or 4+) was similar in both groups (5% and 4% respectively). However, when the stepwise dilatation technique was employed in group II, the incidence had dropped to 2.1%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404937 TI - Myocardial washout of sonicated iopamidol does not reflect the transmural distribution of coronary blood flow. AB - It has been shown in previous studies that myocardial contrast echocardiography provides quantitative information on coronary blood flow. However, the ability of contrast echo to assess the transmural (endo/epicardial) distribution of blood flow is still debated. To test this hypothesis, the left circumflex coronary arteries of six anaesthetized open-chested dogs were cannulated and perfused with blood from the femoral artery. At different rates of coronary blood flow, during adenosine-induced coronary vasodilation, sonicated iopamidol and radionuclide labelled microspheres were injected into the coronary cannula, immediately proximal to a mixing chamber. Two-D echo images were digitized and myocardial time-intensity curves were obtained for the endocardial, mid- and epicardial layers. A good correlation existed between contrast washout of the entire ventricular wall and coronary flow (r = 0.85). However, the washout rate from the endo-, mid- and epicardial layers showed weak correlations with corresponding regional blood flows measured by microspheres (r = 0.56, 0.71 and 0.58, respectively). No significant relationship was found between the endo/epicardial washout ratio and the corresponding flow ratio by microspheres. Thus, measurement of the transmural distribution of coronary blood flow by myocardial contrast echocardiography remains an elusive goal. PMID- 8404938 TI - Diagnostic value of 123I-phenylpentadecanoic acid (IPPA) metabolic and thallium 201 perfusion imaging in stable coronary artery disease. AB - The diagnostic value of 123I-phenylpentadecanoic acid (IPPA) metabolic cardiac imaging was studied in a group (n = 29) of patients with angiographically confirmed CAD using single photon emission computed tomography (SPECT). A symptom limited exercise test was first done with IPPA, and 2 days later with thallium. Medications were not withheld during testing. Fourteen healthy control subjects participated in parallel IPPA and 15 in thallium tests. Data acquisition and output were comparable in the two imaging modalities. By testing various relatively simple criteria for abnormality we found that the semiquantitative interpretation was more accurate than the visual readings. The best compromise of accuracy with the scored criteria consisted of a sensitivity of 86% and a specificity of 86%, obtained with IPPA polar tomograms (mild exercise defect) and a sensitivity of 86% and a specificity of 80% obtained with thallium (regionally decreased washout). With visual interpretation alone, a sensitivity of 83% and a specificity of 71% was detected with IPPA (mild exercise defect) and 72% and 73%, respectively, with thallium (partial reversibility). The sensitivity of the exercise ECG alone was 62%. The results of this study imply that IPPA imaging could be a rational, uncomplicated clinical method for non-invasive diagnosis of CAD. The diagnostic ability of IPPA is at least as good as that of thallium, and it is possible to use them in succession. PMID- 8404939 TI - Hyperventilation-echocardiography test for the diagnosis of myocardial ischaemia at rest. AB - The purpose of this study was to assess the feasibility, safety, specificity and sensitivity of the hyperventilation test performed under echocardiographic monitoring for the provocation of vasospastic ischaemia. Hyperventilation (approximately 30 cycles.min-1 for 5 min) was performed in 104 hospitalized patients, referred for pain typical of angina at rest, under 2-D echocardiographic and 12-lead electrocardiographic monitoring. All the tests were completed and no significant side effect was observed. In-hospital documentation of spontaneous myocardial ischaemia and/or ergonovine-induced ischaemia was achieved in 38 patients (group I). A positive hyperventilation-echocardiography test (occurrence of new transient asynergies or worsening of basal ones) was obtained in 32/38 patients. Among the group I patients, only 23 had diagnostic ST T changes and only 16 experienced chest pain during the hyperventilation-echo test. Of the 66 patients without evidence of myocardial ischaemia at rest (negative ECG monitoring during hospitalization and/or negative ergonovine maleate-echo test)--Group II, none showed echocardiographic changes, seven presented ST-T changes and six complained of typical chest pain during the test. Thus, in relation to in-hospital documentation of myocardial ischaemia at rest, both spontaneous and/or ergonovine-induced episodes, the hyperventilation-echo test showed a specificity of 100%, a sensitivity of 84%, a positive predictive value of 100% and a negative predictive value of 92%. In conclusion, hyperventilation performed under echocardiographic monitoring is feasible and safe; it can be proposed as a screening test to unmask vasospastic myocardial ischaemia in patients with angina at rest, in whom documentation of spontaneous episodes is not available. PMID- 8404940 TI - The ability of QRST isointegral maps to detect myocardial infarction in the presence of simulated left bundle branch block. AB - The clinical value of QRST isointegral maps (I-maps) for the detection of myocardial infarction (MI) in the presence of left bundle branch block (LBBB) was investigated. We recorded I-maps during sinus rhythm and right ventricular (RV) pacing, which simulated LBBB, in 62 patients with MI (42 patients had at least one akinetic segment and the remaining 20 patients had only hypokinesis or normal contraction) and 26 patients without MI. An abnormal decrease in the QRST value of the I-map was assessed by the difference map (D-map), which indicated a '-2SD area', where the QRST integral value was less than the lower limit of the normal range (mean -2SD) calculated from 608 normal individuals. The I-maps recorded during the two activation sequences were similar to each other in patients with and without MI (r = 0.87 and 0.92, respectively). The '-2SD area' was located over the left anterior chest in patients with an anterior MI and over the lower torso in patients with an inferior MI during each activation sequence. We were able to diagnose MI during simulated LBBB with a sensitivity of 84%, a specificity of 81% and a diagnostic accuracy of 83% when we used the criterion that MI is present if the sum of QRST integral values below the normal range (sigma DM) exceeds 100 mV.ms. We were able to diagnose an akinesis with a sensitivity of 81%, a specificity of 85% and a diagnostic accuracy of 83% when we used the criterion that akinesis is present if sigma DM exceeds 500 mV.ms during simulated LBBB.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404941 TI - The value of the QRS scoring system in assessing regional and global left ventricular ejection fraction early after myocardial infarction. AB - In 71 patients with a myocardial infarction (MI) (anterior in 27, inferior in 44 patients) global (GEF) and regional (REF) left ventricular ejection fractions were determined by radionuclide ventriculography and estimated from a 12 lead electrocardiogram (ECG), using Selvester's QRS score, during the early phase of a MI (15 to 21 days following MI). Global ejection fractions determined by radionuclide ventriculography and from ECG using Palmeri's method were: for all MI 40.8 +/- 12.6% vs 39.6 +/- 11.4%; in the group of anterior MI 32.0 +/- 10.0% vs 30.0 +/- 9.7% and in the group of inferior MI 48.9 +/- 12.0% vs 45.1 +/- 8.2%. A good correlation was found between global ejection fractions determined by radionuclide ventriculography and ECG, as well as between radionuclide GEF and ECG score. A weaker correlation was found between radionuclide GEF and enzymes among all MIs and in the group of anterior MI, while in the group of inferior MI this correlation was insignificant. The analysis of REF determined by radionuclide ventriculography and ECG showed the greatest abnormalities in the infarct region, but in the group of anterior MI, dysfunction was present in the whole left ventricle. The comparison of infarct-related REF derived from radionuclide ventriculography, with the QRS score showed a significantly higher correlation than the comparison with enzymes. ECG estimation of REF from a modified Palmeri's equation showed a better correlation with radionuclide REF than did GEF derived from the standard Palmeri's equation: anterior MI; r = 0.90 vs r = 0.82, inferior MI; r = 0.84 vs r = 0.69, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404942 TI - Neurohormonal effects of early treatment with enalapril after acute myocardial infarction and the impact on left ventricular remodelling. AB - Plasma neurohormones were sequentially analysed in 98 patients with acute myocardial infarction randomized to treatment with enalapril or placebo for 4-6 months. Plasma angiotensin converting enzyme activity was rapidly suppressed by enalapril, but unaffected by placebo (P = 0.0001). No significant differences were found in the plasma levels of angiotensin II, aldosterone, atrial natriuretic peptide, noradrenaline, adrenaline or dopamine between the two treatment groups. Among patients with infarct size above median, plasma angiotensin II increased during head-up tilt at one month in the placebo group, but not in the enalapril group. Left ventricular end-diastolic volume (LVEDV) and left ventricular end-systolic volume (LVESV) were evaluated by echocardiography in 28 patients (placebo 15, enalapril 13) and changes in left ventricular volumes between baseline and 4-6 months were calculated. Only in the placebo group was a positive correlation found between plasma levels of noradrenaline at day 5-7 and the subsequent increase in LVEDV (r = 0.78, P = 0.005) and LVESV (r = 0.75, P = 0.008). The same trend was found for angiotensin II, adrenaline and dopamine levels at days 5-7 and the subsequent increase in left ventricular volumes. In the placebo group a negative correlation was found between plasma aldosterone at days 5-7 and the subsequent increase in left ventricular ejection fraction (r = 0.77, P = 0.006) during the study period. Although circulating neurohormones were not significantly influenced by enalapril treatment, it is concluded that enalapril may influence the relationship found between sustained neurohormonal activation and left ventricular remodelling after acute myocardial infarction. PMID- 8404943 TI - The effect of taprostene in patients with acute myocardial infarction treated with thrombolytic therapy: results of the START study. Saruplase Taprostene Acute Reocclusion Trial. AB - Taprostene is a prostacyclin analogue that inhibits platelet aggregation and thus might be a useful adjuvant to thrombolytic agents in acute myocardial infarction. In a placebo-controlled dose rising study, taprostene or placebo was intravenously infused in 80 patients treated with the thrombolytic agent saruplase (rscu-PA) for acute myocardial infarction. Three doses of taprostene were used: 6.25; 12.5; or 25.0 ng.kg-1 x min-1. Taprostene or placebo was infused for 48 h, followed by a 24 h tapering period. All 80 patients had short symptom to-treatment delay and marked ST segment elevation. Patency at 90 min was documented in 58/78 patients (two patients had no angiography). Success rate varied from 67-82% in the four treatment arms (P = 0.33). Patency after rescue PTCA was seen in 10 out of 13 patients. Of the 58 patients having a patent artery at 90 min, none of the 43 taprostene patients and one of the 15 placebo patients had a re-occluded artery at the second angiography at 32-48 h (5/58 patients had no recatheterization). Conversely, of nine patients who had successful rescue PTCA, three of four placebo patients had a re-occluded artery at the second angiography compared to one of five taprostene patients (one placebo patient had no recatheterization) (P = 0.33). Safety evaluation revealed no major difference between the placebo plus saruplase and the taprostene plus saruplase groups. Taprostene was well tolerated up to 25 ng.kg-1 x min-1. Although taprostene did not affect 90 min patency, there was a trend to better maintenance of patency after rescue PTCA. PMID- 8404944 TI - Comparison of flecainide and procainamide in cardioversion of atrial fibrillation. AB - In this prospective, controlled and randomized cross-over study we tried to establish the efficiency and safety of flecainide vs procainamide for the treatment of acute atrial fibrillation. Eighty patients (30 females, 50 males, mean age: 55 +/- 14 years) were included. Patients entered into the study if they had atrial fibrillation of recent onset (< 24 h) with a ventricular rate > 100 beats.min-1 at rest and were < 75 years of age. Exclusion criteria were any sign of heart failure, conduction disturbances, sick sinus syndrome or acute ischaemic events. Randomly 40 patients received flecainide and 40 procainamide as the first treatment. There were no significant clinical difference between the two groups. Procainamide ws given at a dose of 1 g infused over 30 min, and followed by an infusion of 2 mg.min-1 over 1 h. Flecainide was given at a dose of 1.5 mg.kg-1 over 15 min followed by an infusion of 1.5 mg.kg-1 over 1 h. Drug infusion was continued until maximal dose, intolerance or reversion to sinus rhythm. After 1 h of wash out, patients remaining in atrial fibrillation were started on the second drug. Left atrial size was measured by echo. Serum levels of drug and atrial size did not differ between patients who returned to sinus rhythm and those who remained in atrial fibrillation. Conversion to sinus rhythm was achieved in 37 (92%) of the 40 patients treated with flecainide and 25 (65%) of those treated with procainamide (P < 0.001). The time required for reversion to sinus rhythm was similar between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404945 TI - Induction and the possible mechanism of ventricular tachycardia after catheter ablation with direct current shocks in dogs. AB - Cathodal DC shocks (150-J) were administered via the His bundle to 20 closed chest dogs, and in a further three dogs 25-J cathodal shocks were given via the left ventricular endocardium. In 18 dogs, including three that underwent left ventricular ablation, Holter electrocardiograms were recorded from 1 to 7 days after ablation, and 4 weeks after ablation. There were frequent episodes of sustained ventricular tachycardia (VT) from the first few hours after ablation to 4 days after ablation in all dogs, but both the rate and the coupling interval of VT were variable. In five conscious dogs stimulated 1 day after ablation, it was difficult to induce and terminate VT repeatedly. There was a direct relationship between the paced cycle length and the interval of the last paced beat to the initiating VT beat in three out of four dogs. In the fourth dog there was an inverse relationship. There was no transient entrainment with ventricular burst pacing during VT in any of the four dogs tested. The effects of lidocaine (2-3 mg.kg-1), verapamil (0.2-0.4 mg.kg-1), and propranolol (0.2 mg.kg-1) on VT were tested within 2 days of ablation in 10 conscious dogs. In general, both lidocaine and verapamil terminated VT, and propranolol slowed VT. In conclusion, VT soon after ablation possibly results from triggered activity, although abnormal automaticity cannot be ruled out. PMID- 8404946 TI - Aortic valve replacement four years after cardiac transplantation. AB - Severe aortic insufficiency developed in a 56-year-old woman 4 years after cardiac transplantation. Aortic valve replacement was performed with insertion of a 23 mm Carbo Medics mechanical prosthesis. The patient recovered completely and is, 5 months after valvular replacement, in New York Heart Association functional class I. PMID- 8404947 TI - Malignant lymphoma in the donor heart after heart transplantation. AB - A case of post-transplant lymphoproliferative disease (PTLD) with donor heart involvement is reported. The 49-year-old patient presented with heart failure initially ascribed to acute graft rejection. The treatment with high doses of immunosuppressive agents was unsuccessful and the outcome rapidly fatal. This case suggests that cardiac failure occurring after high doses of immunosuppressive therapy could be a sign of early PTLD in heart transplant recipients. PMID- 8404948 TI - Growth rate of primary left atrial myxoma. PMID- 8404949 TI - Value of intravascular ultrasound in the diagnosis and characterization of patent ductus arteriosus in an adult patient. AB - We report the case of a 47-year-old patient with a patent ductus arteriosus who underwent angiography and percutaneous intravascular ultrasound examination. The case illustrates that intravascular ultrasound contributes to the characterization of patent ductus arteriosus when a surgical or trans-catheter closure is planned, as calcifications of the wall as well as aneurysmal dilatations can be ruled out. Furthermore, measurements of the diameter of the ductus can be made. PMID- 8404950 TI - Radiopharmaceuticals and the single European market. PMID- 8404951 TI - A method for stabilising technetium-99m exametazime prepared from a commercial kit. AB - Technetium-99m exametazime (99mTc-d,l-HMPAO), prepared by the reconstitution of the Ceretec kit, is widely used in the investigation of regional cerebral blood flow. The radiochemical purity specification of not less than 80% lipophilic complex requires that the kit is used within 30 min of reconstitution. In certain circumstances this imposes restrictions on its clinical availability. A number of approaches to extending the 30-min shelf life have been proposed and these are discussed. A method of stabilising the kit by the addition of 200 micrograms cobalt chloride hexahydrate (CoCl2 x 6H2O) in 2 ml of water is described. The addition of this solution can extend the shelf life of the reconstituted kit to at least 5 h post reconstitution. PMID- 8404952 TI - Technetium-99m labelled LDL as a tracer for quantitative LDL scintigraphy. I. Tracer purification, in vitro and in vivo long-term stability, in vitro validation and biodistribution. AB - The goal of the present study was to optimize technetium-99m labelling of low density lipoprotein (LDL) and to investigate the in vitro and in vivo properties of the tracer to determine whether its application for quantitative scintigraphy of hepatic LDL receptor activity is feasible. LDL labelled with iodine-125 by the iodine monochloride method was used as a reference tracer. Comparison of different assessments of radiochemical purity [trichloro-acetic acid precipitation (%ppTCA), paper chromatography, size-exclusion chromatography and chloroform-methanol extraction] exhibited %ppTCA to be superior as a parameter of tracer quality. In spite of a high radiochemical purity immediately after labelling, modifications of 99mTc labelling of LDL did not overcome the poor long term stability of the tracer. Subsequent dialysis in phosphate buffer over about 3 h sufficiently increased the long-term stability in vitro and in vivo. The competitive recognition of dialysed 99mTc-LDL and 125I-LDL with native LDL by high-affinity binding sites was demonstrated in human hepatoma cells (HepG2) and human fibroblasts. Biodistribution data of simultaneously injected 99mTc-LDL and 125I-LDL in New Zealand White rabbits showed a high uptake of both tracers in tissues with high LDL receptor activity, yet 99mTc-LDL uptake exceeded 125I-LDL uptake by two- to sevenfold. In contrast to 125I-LDL, 99mTc-LDL showed a higher unspecific uptake into the bone marrow and the spleen, suggesting an additional uptake mechanism probably via the scavenger pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404953 TI - Technetium-99m labelled LDL as a tracer for quantitative LDL scintigraphy. II. In vivo validation, LDL receptor-dependent and unspecific hepatic uptake and scintigraphic results. AB - The purpose of this study was to determine whether the hepatic uptake of dialysed technetium-99m labelled low-density lipoprotein (99mTc-LDL) reflects the hepatic LDL receptor activity and to what extent the non-LDL receptor-dependent 99mTc-LDL uptake by non-parenchymal cells relates to the diagnostic utility of quantitative 99mTc-LDL scintigraphy of the liver. New Zealand White rabbits and Watanabe Heritable Hyperlipidaemic rabbits, which were sacrificed 24 h after simultaneous injection of 99mTc-LDL and iodine-125 labelled LDL, were clearly discriminated by their hepatic 99mTc-LDL uptake according to their genetically different hepatic LDL receptor activity. Yet the hepatic 99mTc-LDL uptake exceeded the 125I-LDL uptake in all animals. The different hepatic uptake of the tracers was elucidated in the isolated perfused rat liver and was due to rapid intracellular degradation and the release of low molecular catabolites of 125I-LDL. In contrast, 99mTc activity was trapped in the liver. Analysis of biliary 99mTc activity provided evidence for the excretion of 99mTc-labelled apolipoprotein B. The amount of biliary excreted protein-bound 99mTc was linked to total hepatic 99mTc-LDL uptake and presumably reflected LDL receptor-mediated apolipoprotein excretion. Collagenase liver perfusion in Sprague-Dawley rats 90 min following simultaneous injection of 99mTc- and 125I-LDL and subsequent cell separation by gradient centrifugation revealed that 99mTc-LDL and 125I-LDL had a comparably low uptake into non-parenchymal cells; thus its contribution can be neglected for scintigraphic purposes. Planar scintigraphy was performed in New Zealand White and Watanabe Heritable Hyperlipidaemic rabbits.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404954 TI - Validation of quantitative brain dopamine D2 receptor imaging with a conventional single-head SPET camera. AB - Phantom measurements were performed with a conventional single-head single-photon emission tomography (SPET) camera in order to validate the relevance of the basal ganglia/frontal cortex iodine-123 iodobenzamide (IBZM) uptake ratios measured in patients. Inside a cylindrical phantom (diameter 22 cm), two cylinders with a diameter of 3.3 cm were inserted. The activity concentrations of the cylinders ranged from 6.0 to 22.6 kBq/ml and the cylinder/background activity ratios varied from 1.4 to 3.8. From reconstructed SPET images the cylinder/background activity ratios were calculated using three different regions of interest (ROIs). A linear relationship between the measured activity ratio and the true activity ratio was obtained. In patient studies, basal ganglia/frontal cortex IBZM uptake ratios determined from the reconstructed slices using attenuation correction prior to reconstruction were 1.30 +/- 0.03 in idiopathic Parkinson's disease (n = 9), 1.33 +/- 0.09 in infantile and juvenile neuronal ceroid lipofuscinosis (n = 7) and 1.34 +/- 0.05 in narcolepsy (n = 8). Patients with Huntington's disease had significantly lower ratios (1.09 +/- 0.04, n = 5). The corrected basal ganglia/frontal cortex ratios, determined using linear regression, were about 80% higher. The use of dural-window scatter correction increased the measured ratios by about 10%. Although comprehensive correction methods can further improve the resolution in SPET images, the resolution of the SPET system used by us (1.5-2 cm) will determine what is achievable in basal ganglia D2 receptor imaging. PMID- 8404955 TI - Quantification of technetium-99m hexamethylpropylene amine oxime brain uptake in routine clinical practice using calibrated point sources as an external standard: phantom and human studies. AB - Quantitative methods for calculation of regional cerebral blood flow with technetium-99m hexamethylpropylene amine oxime (99mTc-HMPAO) have been proposed. These methods are very labour intensive and therefore are not useful in routine clinical practice. We describe a simple alternative method, using calibrated point sources as a scaling factor, whereby the tomographic slices are displayed as regional 99mTc-HMPAO brain uptake per cm3 brain tissue in 10(-6) of the injected lipophilic dose. The method was validated on Jaszczak and Hoffman phantoms using a three-detector system with HR parallel and HR fan-beam collimators. Under the optimal conditions described in this paper, the measured to real activity ratio was 1.00 (SD = 0.06). The reproducibility of the cerebellar uptake in a group of ten normal volunteers and five patients was studied. Intra-individually a mean deviation of 12.6% was observed for the total group. For those persons with a heart rate difference of less than 5 units between the two studies, a mean deviation of 7.2% was obtained. Quantitative 99mTc-HMPAO brain uptake images can be useful for longitudinal studies, especially for follow-up, activation and pharmacological studies. PMID- 8404956 TI - Radioimmunoscintigraphy with technetium-99m labelled monoclonal antibody, 1A3, in colorectal cancer. AB - This study of radioimmunoscintigraphy (RIS) was primarily undertaken to determine how the imaging results related to surgical findings. Technetium-99m radiolabelled 1A3, a monoclonal antibody against a columnar cell surface antigen, was used. No adverse effects or thyroid uptake was observed in 127 studies. The 85 primary colorectal cancers were all image positive. In the assessment of recurrent tumour in the abdomen or pelvis, the accuracy was 33/35 (94%), including true-positive findings in some whose serum carcinoembryonic antigen was normal. There was a positive predictive value for abdominal or pelvic recurrence of 92% and a negative predictive value of 100%, at a prevalence of 66%. In those patients whose liver was able to be evaluated, the accuracy was 72/79 (91%). There was a positive predictive value for liver metastases of 88% and a negative predictive value of 93%, at a prevalence of 32%. The simple procedure for labelling antibody with 99mTc and its ready availability allow a completed report to be given within 24 h of the request. PMID- 8404957 TI - Hypertension in paediatrics: can pre- and post-captopril technetium-99m dimercaptosuccinic acid renal scans exclude renovascular disease? AB - In children over 1 year of age, renal disease is the commonest cause of hypertension. Arteriography is considered the reference method to establish the diagnosis of renovascular disease; however, it is an invasive technique with a high radiation burden for children. This was a retrospective study of pre- and post-capto-technetium-99m dimercaptosuccinic acid (DMSA) imaging compared with arteriography in 18 children between the ages of 3 and 17 years. Alone, the 99mTc DMSA scan is a sensitive indicator of renal parenchymal disease, although non specific in suggesting the pathology. The combination of pre- and post-captopril studies may increase the sensitivity and specificity in the diagnosis of renovascular disease in the presence of hypertension. This work suggests that a screening investigation with a low radiation burden can be carried out at most institutions; if the investigation is positive, there will be a high index of suspicion that renovascular disease is the cause of the hypertension. PMID- 8404958 TI - Bone single-photon emission tomography in recent meniscal tears: an assessment of diagnostic criteria. AB - Bone single-photon emission tomography (SPET) was performed in 40 patients within 6 months of acute knee injury where internal derangement of the knee was suspected, and the results related to the arthroscopy findings. Scan features with high sensitivity, specificity, and predictive accuracy for a meniscal tear could not be obtained on planar imaging. However, a half-crescent or more of increased tibial plateau activity on transaxial SPET gave a sensitivity of 89%, a specificity of 76%, a positive predictive accuracy of 77% and a negative predictive value of 89%. For longitudinal (bucket handle) tears alone the optimum scan pattern was a full crescent of increased tibial plateau activity with adjacent femoral activity and increased blood pool activity which gave corresponding values of 78%, 94%, 78% and 93%. It is concluded that the inclusion of tibial plateau activity of less than a full crescent and the presence of femoral condyle and blood pool activity in the diagnostic criteria improves the ability of bone SPET to detect meniscal tears. The value of bone SPET in the diagnosis of meniscal tears suggests that it could have a significant role to play in the management of knee injuries. PMID- 8404959 TI - Accuracy of a rapid 10-minute carbon-14 urea breath test for the diagnosis of Helicobacter pylori-associated peptic ulcer disease. AB - Urease in the human gastric mucosa is a marker for infection with Helicobacter pylori (HP), an organism which is associated with peptic ulcer disease. To detect gastric urease, we examined 184 patients (144 males, 40 females; mean age: 49.8 +/- 15.6 years) with suspected peptic ulcer disease. Fasting patients were given orally 5 microCi of carbon-14 labelled urea. From each patient only one breath sample was collected in hyamine at 10 min. The amount of 14C collected at 10 min was expressed as follows: [(DPM/mmol CO2 collected)/(DPM administered)] x 100 x body weight (kg). The presence of HP colonization was determined by examination of multiple endoscopic prepyloric antral biopsy specimens subjected to culture or a rapid urease test. For the purpose of this study, HP-positive patients were defined as those with characteristic bacteria as indicated by a positive result of either the culture or the rapid urease test; HP-negative patients were defined as those with negative findings on both the culture and the rapid urease test. Of the 184 cases, 99 (53.8%) were positive for HP infection, and 85 (46.2%), negative. The sensitivity and specificity of the rapid 10 min 14C-urea breath test for the diagnosis of HP-associated peptic ulcer disease were evaluated by a receiver operating characteristic (ROC) curve with a variable cut-off value from 1.5 to 4.5. When a cut-off value of 1.5 was selected, the sensitivity was 100% and the specificity, 83.5%; when a cut-off value of 4.5 was selected, the sensitivity was 54.5% and the specificity, 97.6%. PMID- 8404960 TI - EEC directives and guidelines applicable to radiopharmaceuticals--1993. Status quo with particular emphasis on the registration of radiopharmaceuticals which were on the market prior to January 1992. AB - The manufacture, scale and supply of radiopharmaceuticals in the EEC is regulated by directives that are incorporated into the national laws of the member states. The situation as of 1 January 1993 was not too optimistic, however, as the processing of licensing applications had been completely misjudged. Not one product had been registered as of 1 January. The costs involved are also high and since the European market for radiopharmaceuticals is relatively small, the market cannot afford this. It would appear that the EEC directives are inadequate and too non-specific, so revision is indicated. PMID- 8404962 TI - Effective dose from radiopharmaceuticals. PMID- 8404961 TI - Somatostatin receptor scintigraphy with [111In-DTPA-D-Phe1]- and [123I-Tyr3] octreotide: the Rotterdam experience with more than 1000 patients. AB - Various tumours, classically specified as either neuroendocrine or non neuroendocrine, contain high numbers of somatostatin receptors, which enable in vivo localization of the primary tumour and its metastases by scintigraphy with the radiolabelled somatostatin analogue octreotide. In addition granulomas and autoimmune processes can be visualized because of local accumulation of somatostatin receptor-positive activated mononuclear leucocytes. In many instances a positive scintigram predicts a favourable response to treatment with octreotide. It is tempting to speculate that octreotide labelled with an appropriate radionuclide might be used in cancer therapy. The successful application of radiolabelled octreotide in scintigraphy indicates the possible usefulness of other radiolabelled peptides, either native peptides or derivatives of these, in, for example, nuclear oncology. The small size of these peptides, e.g. bombesin and substance P, is of the utmost importance for a relatively fast blood clearance, thus leading to low background radioactivity. In this way peptides are powerful alternatives to (fragments of) monoclonal antibodies, the application of which to scintigraphic localization of specific cell surface antigen-bearing tumours is plagued by slow blood clearance and, hence, high background levels. PMID- 8404963 TI - The pioneers of pediatric medicine. Sir Archibald Garrod (1857-1936). PMID- 8404964 TI - Otoacoustic emissions: a new method to diagnose hearing impairment in children. AB - Otoacoustic emissions (OAEs) are epiphenomena of sensitive, amplifying processes during hearing which can be detected in persons with normal inner ear function. They originate from the cochlea and are interpreted as an energy leakage of cochlear processes, perhaps resulting from active outer hair cell movements. OAEs travel from the cochlea through the middle ear to the external auditory canal where they can be detected using sensitive miniature microphones. Transient evoked (TEOAE) tests allow to otoacoustic emissions non-invasively check the integrity of the cochlea. In the neonatal period, registration of OAEs can be accomplished during natural sleep. In infants and neonates TEOAEs can be used as screening test with a screening level at 30 dB HL in paediatric audiology. They are less time consuming and elaborate than auditory brainstem responses (ABR) and they are more sensitive than behavioral tests. TEOAEs are constant over long periods of time and they are reduced or absent due to various adverse influences in the inner ear. These latter characteristics may allow monitoring of the inner ear function over time e.g. during disease and/or during ototoxic therapeutic interventions. Limitations of this new method are due to the fact that TEOAEs are absent in patients with a more than 30 dB HL hearing loss. Thus a hearing threshold cannot be determined. Diseases of the inner ear which are common in early childhood (like otitis media) reduce the transfer of TEOAEs and may wrongly indicate a cochlear hearing disorder. New methods for evaluation and interpretation of TEOAE test results are currently developed which may allow to circumvent this problem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404965 TI - Glycaemic effect and satiating capacity of potato chips and milk chocolate bar as snacks in teenagers with diabetes. AB - In 14 adolescents with diabetes, the postprandial blood glucose, after ingestion of two popular snacks, milk chocolate bar and potato chips, was compared to a prescribed mid-afternoon snack that was isocaloric with the comparison snacks. The prescribed diabetes snack consisted of wholemeal wheat bread, margarine, cheese and apple (1533 kJ/365 kcal): 14% protein, 32% fat and 54% carbohydrate. The milk chocolate bar and the potato chips contained 4% protein, 55% fat and 41% carbohydrate. The mean blood glucose peak was 4.7 (+/- 0.8) mmol/l after the regular diabetes snack, after the milk chocolate bar 2.9 (+/- 0.6) mmol/l, and 3.2 (+/- 0.6) mmol/l after the potato chips (P > 0.05). The mean incremental area under the blood glucose curve was 450.3 (+/- 105.5) mmol/l x 180 min for the regular diabetes snack, 269.5 (+/- 96.7) mmol/l x 180 min for the milk chocolate bar and 191.7 (+/- 95.0) mmol/l x 180 min for the potato chips (P > 0.05). We conclude that an occasional exchange of a regular diabetes afternoon snack for an isocaloric amount of milk chocolate bar or potato chips has no negative impact on the postprandial blood glucose. PMID- 8404966 TI - Cow's milk protein intolerance in infants under 1 year of age: a prospective epidemiological study. AB - Incidence and clinical manifestation of cow's milk protein intolerance (CMPI) were studied in 1158 unselected newborn infants followed prospectively from birth to 1 year of age. No food changes were required in 914 infants who were used as healthy controls. When CMPI was suspected (211 infants), diagnostic dietary interventions according to a standard protocol were performed. After exclusion of lactose intolerance, two positive cow's milk elimination/challenge tests were considered diagnostic of CMPI. Two hundred and eleven symptomatic infants were examined for possible CMPI. A large group of 80 infants improved on a lactose reduced formula. In 87/211 infants CMPI was excluded (sick controls). Finally CMPI was proven in 26 infants. The calculated incidence rate for CMPI was 2.8%. The principal symptoms in infants with CMPI were gastrointestinal, dermatological and respiratory in 50%, 31% and 19% respectively. A positive family history for atopy (first or second degree relatives) was more frequent in either CMPI infants (65%), or sick controls (63%) when compared to either healthy controls (35%) or infants improving on a low lactose formula (51%). Differences between patients with CMPI and sick controls were only found for the presence of atopy in at least 2 first degree relatives [(5/26 in CMPI infants and 4/87 in sick controls (P < 0.05)] and for multiorgan involvement [10/26 infants with CMPI as opposed to 12/87 in the sick control group (P < 0.02)]. These statistical differences are too weak to be of clinical value. PMID- 8404967 TI - Chorea as a manifestation of rheumatic fever--a 30-year survey (1960-1990). AB - Sydenham chorea, a major manifestation of acute rheumatic fever, has been the most common form of acquired chorea during childhood. Despite the recent dramatic decline in both incidence and severity of rheumatic fever in our area, the frequency of carditis was unchanged. This study investigated retrospectively the incidence of chorea in the last three decades (1960-1990) in our area. During the 30 years of the survey, 28 patients with Sydenham chorea were treated in our centre of whom 10 were seen between 1960-1970, 17 between 1970-1980, and only one patient between 1980-1990. PMID- 8404968 TI - Increased levels of urinary interleukin-6 in Kawasaki disease. AB - Kawasaki disease (KD) often presents with abnormal urinary findings, such as aseptic pyuria, mild proteinuria and microscopic haematuria. In this study, we measured urinary interleukin-6 (IL-6) by a sensitive sandwich ELISA assay using mouse monoclonal antibodies against recombinant IL-6 to elucidate the role of IL 6 in the pathogenesis of renal lesions in KD. Serum IL-6 levels were increased in acute KD as well as in febrile controls. Importantly, urinary IL-6 levels were consistently elevated in patients with acute KD, but much lower in febrile controls. Urinary IL-6 levels returned steadily to normal during the convalescent phase. In addition to IL-6, urinary levels of N-acetyl-beta-D-glucosaminidase (NAG) and beta 2-microglobulin (beta 2-mg) were also elevated during the acute phase of this disease. Eosinophils and macrophages were identifiable in urinary sediments from these patients. The increased levels of urinary IL-6 in combination with increased NAG and beta 2-mg seemed to suggest the presence of certain renal parenchymal lesions with cellular infiltration during the acute phase of the disease. IL-6 may serve as clinically useful parameter for the detection and monitoring of the renal involvement in KD. PMID- 8404969 TI - Differential diagnosis of hyperphenylalaninaemia by a combined phenylalanine tetrahydrobiopterin loading test. AB - We describe a new fully reliable method for the differential diagnosis of tetrahydrobiopterin-dependent hyperphenylalaninaemia (HPA). The method comprises the combined phenylalanine (Phe) plus tetrahydrobiopterin (BH4) oral loading test and enables the selective screening of BH4 deficiency when pterin analysis is not available or when a clear diagnosis has not been previously made. It should be performed together with the measurement of dihydropteridine reductase (DHPR) activity in blood. The new combined loading test was performed in nine patients with primary HPA, three with classical phenylketonuria (PKU), three with DHPR deficiency, and three with 6-pyruvoyl tetrahydropterin synthase (PTPS) deficiency. Three hours after oral Phe loading (100 mg/kg body weight), synthetic BH4 was administered orally at doses of either 7.5 or 20 mg/kg body weight. Amino acid (Phe and tyrosine) and pterin (neopterin and biopterin) metabolism and kinetics were analysed. By exploiting the decrease in serum Phe 4 and 8 h after administration, a clear response was obtained with the higher BH4 dose (20 mg/kg body weight), allowing detection of all cases of BH4 deficiency, as well as differentiation of BH4 synthesis from regeneration defects. Since DHPR deficient patients who were previously shown to be non-responsive to the simple BH4 loading test gave a positive response, the combined Phe plus BH4 loading test can be used as a more reliable tool for the differential diagnosis of HPA in these patients. Moreover, it takes advantage of being performed while patients are on a Phe restricted diet. PMID- 8404970 TI - Molybdenum cofactor deficiency in two siblings: diagnostic difficulties. AB - Two siblings with molybdenum cofactor deficiency are presented. They showed clinical, biochemical and neuroradiological features very similar to those of the few previously described cases. Difficulties in diagnosis are emphasised. PMID- 8404971 TI - Ocular involvement in two symptomatic congenital erythropoietic porphyria. AB - Congenital erythropoietic porphyria (Gunther disease, CEP) is a rare autosomal recessive disorder of haeme biosynthesis. It is characterized by extreme photosensitivity and the excretion of large amounts of uroporphyrin I and coproporphyrin I in the urine and coproporphyrin I in the faeces. We have diagnosed two cases of congenital erythropoietic porphyria, who were first cousins once removed. They had recurrent skin bullae, scarring on the face and hands, hirsutism, discoloured fluorescent teeth, red urine, increased haemolysis and grossly increased excretion of porphyrin. Both children had blepharitis and their sclera gave pink fluorescence under long wave ultraviolet light, mainly in the interpalpebral fissures. All the features of our two patients, except the ocular lesions, conformed to cases of CEP reported in the literature. We have encountered no other reports on ocular lesions in CEP since first described by Chumbley in 1977. PMID- 8404972 TI - Effective renal plasma flow in patients with glycogen storage disease type I. AB - To evaluate effective renal plasma flow (ERPF) we performed renal scintigraphies with 99mTc-Mercaptoacetyl-triglycine (MAG3) in nine patients with glycogen storage disease I (GSD I) (age: 16 +/- 7 years). Two patients presented with proteinuria, none showed hyperaminoaciduria, disturbed tubular reabsorption of phosphate or hypertension. 99mTc-MAG3 clearance values were elevated in eight out of nine patients (865 +/- 233 ml/min/1.73 m2 body surface area) and exceeded the age-dependent mean values by 21%-145%. ERPF values in patients with poor metabolic control were higher than in patients with long-term good metabolic control (988 +/- 186 vs. 619 +/- 55 ml/min/1.73 m2; P < 0.05). We conclude that enhanced ERPF is a common finding in GSD I patients, which precedes clinically overt nephropathy. Renal scintigraphy with 99mTc MAG3 is a suitable method for the early detection and monitoring of kidney dysfunction in GSD I. PMID- 8404973 TI - Reduction in the risk of bronchopulmonary dysplasia from 1980-1990: results of a multivariate logistic regression analysis. AB - A retrospective analysis (1980-1990) of normally formed low birthweight (< 2500 g) infants surviving to at least 28 days following intermittent positive pressure ventilation (IPPV) for longer than 12 h was performed. Bronchopulmonary dysplasia (BPD) was defined as oxygen dependency at 28 days with characteristic radiographic findings. Logistic regression analysis of risk factors, before and after the initiation of IPPV was performed on 412 infants. Decreasing birth weight (BW) and gestational age (GA) were associated with an increased risk of BPD. When controlled for these variables, predictive factors prior to IPPV were gender, age at IPPV, respiratory diagnosis, and year of birth. Following IPPV, duration of peak inspiratory pressure > 25 cm H2O, duration of fraction of inspired oxygen (FiO2) > 0.60 (DO2), maximum peak inspiratory pressure (MPIP), maximum FiO2, patent ductus arteriosus, bacteraemia and either pneumothorax or pulmonary interstitial emphysema were associated with an increased risk of BPD. Adjusting for BW and GA, there was a significant reduction in BPD risk from 1980 1990 (relative odds of 0.88 for each year compared to the previous year). This trend could be largely accounted for by decreases in MPIP and DO2 during the study period. Surfactant treatment was not independently associated with a significant change in the risk of BPD. Based on this analysis, we developed a scoring system for predicting the risk of BPD in the neonatal period which we evaluated in a random sample of infants. This predicted infants at risk of BPD with a sensitivity of 65% and a specificity of 88%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404974 TI - Simultaneous recording of brainstem and cortical acoustic evoked potentials in children: methodical aspects and normative data. AB - Early and late acoustic evoked responses were simultaneously recorded from 42 children aged 3-17 years by means of a computer-based device using self-developed software. Filling the interstimulus intervals of late acoustic evoked potentials with clicks leads to an increase of latencies and a decrease of amplitudes of N1 and P2 components. Age-dependent normal values for various recording conditions are given. PMID- 8404975 TI - An assessment of the microsocial environment of children diagnosed as "sudden infant death" using the "process" inventory. AB - An assessment of the microsocial environment of infants dying unexpectedly was carried out using the Paediatric Review and Observation of Children's Environmental Support and Stimulation (PROCESS) inventory. We studied 48 families where the diagnosis of sudden infant death was confirmed by full necropsy and who died in St. petersburg from 1986 to 1991. A control group consisted of an equal number of age, sex and observer matched live controls from the same city. The infants presenting as unexpected deaths had significantly lower estimates of developmental stimulation, organisation, total parent questionnaire and clinical observation scores. The total PROCESS score values were also lower in the families with sudden infant death than in the controls. A defect in developmental stimulation may be a contributory factor in some babies presenting as unexpected infant deaths. PMID- 8404976 TI - Hepatic tumours during androgen therapy in Fanconi anaemia. AB - The occurrence of liver tumours in the course of Fanconi anaemia (FA) has been well documented. We present a case, review the literature and conclude that androgen therapy would increase the risk of developing tumours, most of which appear to be benign (adenomas or peliosis) and androgen-dependent, generally decreasing in size after cessation of treatment. Survival of patients is poor, mostly because of the rapid evolution of the tumour. In the absence of an allogenic bone marrow transplantation, administration of haematopoietic growth factors might be effective. As a preventive measure, other types of unsubstituted androgens may be used. PMID- 8404977 TI - Myocardial fibrosis--a rare complication in patients with cystic fibrosis. AB - We report a 10-month-old male infant who was admitted to our hospital with a history of failure to thrive and bulky stools. On examination, he was dystrophic and had a protruding abdomen, but he was well oxygenated and his lungs were clear on auscultation. A tachycardia of 145 beats per min and radiological evidence of cardiomegaly indicated involvement of the heart, but an ECG failed to show signs of myocarditis or cardiac hypertrophy. An elevated sweat chloride concentration of 141 mEq/l confirmed the diagnosis of cystic fibrosis (CF). Molecular analysis revealed heterozygosity for the common mutation delta F508. He died unexpectedly of a sudden cardiac arrest 2 days later. Autopsy revealed scattered myocardial necrosis and fibrosis. Some 50 documented cases of myocardial fibrosis in infants with CF have been reported. Suggested causes such as malnourishment and hypovitaminosis remain speculative as systematic studies have yet to be done. PMID- 8404978 TI - Diagnostic difficulties and positive therapeutic response in a patient with sinus histiocytosis with massive lymphadenopathy. PMID- 8404979 TI - Severe endophthalmitis after neonatal skin lesions with positive cultures of Aspergillus fumigatus. PMID- 8404980 TI - Vancomycin intoxication in a preterm neonate. PMID- 8404981 TI - Benign infantile familial convulsions. PMID- 8404982 TI - Food-induced pruritus ani: a variation of allergic target organ? PMID- 8404983 TI - Dipylidium caninum in an infant. PMID- 8404984 TI - Tri-country Meeting on Election Microscopy. Zurich, 5-11 September 1993. Abstracts. PMID- 8404985 TI - Effect of hyperbaric oxygen and surgery on experimental multimicrobial gas gangrene. AB - An experimental multimicrobial gas gangrene rat model was developed and the therapeutic effect of surgery was compared to the combined effect of surgery and hyperbaric oxygen (HBO). The infection was caused by an intramuscular injection of a mixture of bacteria including Clostridium perfringens, Bacteroides fragilis, Escherichia coli and Streptococcus faecalis. The mortality, morbidity and wound healing were assessed during a follow-up of 2 weeks. The mortality of the control rats was 60%, with rats treated with surgery alone 35% and the combined treatment group 20% (NS). In the combined treatment group, 84.4% of the survivors healed completely; the corresponding figure in the surgery group was 15.4% (p < 0.001). In experimental multimicrobial gas gangrene the addition of HBO to surgery reduces morbidity and improves wound healing but does not reduce mortality statistically significantly. PMID- 8404986 TI - Histomorphological evaluation of wound healing of rabbit oviduct after microsurgical reanastomosis with the use of autologous fibrin adhesive, human fibrin adhesive or poly-glycolic acid suture. AB - The morphology of the healing process of microsurgical reanastomosis of the rabbit oviduct with the use of fibrin adhesive, autologous and heterologous, and conventional sutures is described. Both oviducts in 48 rabbits were cut and reanastomosis were performed. The rabbits were killed at different intervals after the operations, ranging from 2 h to 28 days, and the anastomoses were evaluated by histomorphological examination. The autologous fibrin adhesive was absorbed after a week and an uncomplicated healing was observed. Heterologous fibrin adhesive caused a granulomatous inflammation interpreted as an immune reaction of the host to the foreign protein, and conventional suturing resulted in severe tissue damage with an intensive inflammatory reaction. PMID- 8404987 TI - Role of the hepatovasculature in free radical mediated reperfusion damage of the liver. AB - This study was undertaken in order to assess the role of purely circulation related effects upon free-radical-mediated reperfusion injury in the liver by comparing the respective effects of the oxygen free-radical scavenger superoxide dismutase (SOD) and the vasodilative action of papaverine in an ischemia/reperfusion model of the liver. Livers from male Wistar rats were rinsed blood free via the portal vein and stored ischemically (60 min at 37 degrees C in Krebs-Henseleit solution and 60 min at 4 degrees C in Euro-Collins solution). Reperfusion was carried out at a constant flow of 30 ml/min for 45 min at 37 degrees C in a nonrecirculating manner. Warm ischemic damage was evident in untreated livers compared to control livers, submitted solely to cold ischemia for 2 h at 4 degrees C, by increased vascular resistance upon reperfusion, enhanced enzyme leakage from the parenchyma (glutamate pyruvate transaminase, glutamate dehydrogenase) and from the endothelium (purine-nucleoside phosphorylase), reduced tissue content of ATP and enhanced lipid peroxidation. Preischemic treatment with SOD or papaverine (the latter also given during reperfusion) significantly reduced hepatic vascular resistance and parenchymal enzyme loss in a comparable manner. Both drugs resulted in a significant increase of hepatic tissue content of ATP at the end of reperfusion. SOD, but not papaverine, prevented the leakage of purine-nucleoside phosphorylase and significantly reduced the tissue levels of lipid peroxides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404988 TI - Orthotopic liver transplantation in the rat: comparison of models with and without rearterialization of the graft. AB - A total of 112 orthotopic liver transplants with (AOLT) and without (NOLT) rearterialization were performed in 2 series using male Lewis rats as donors and recipients. In the first series, the 2-week survival rates were 5/11 (45.5%) in the NOLT group and 22/31 (71%) in the AOLT group. The difference in survival was due to operative experience, as the NOLT transplants were performed before the AOLT transplants. In the second series, the NOLT (n = 30) and AOLT transplants (n = 40) were performed in random order. No differences in 2-week survival rates were found [NOLT, 29/30 (96.7%); AOLT, 37/40 (92.5%)]. Recovery of pre-operative body weight was, however, faster in the AOLT (7 days) than in the NOLT group (14 days). In the NOLT group, elevation of serum levels of GOT (p < 0.05 vs. AOLT at 3 days) and alkaline phosphatase (p < 0.01 vs. AOLT at 3 days and 1 week; p < 0.05 at 4 weeks) were found. We conclude from our results that in OLT without prolonged graft storage, survival is a sensitive marker of operative success only in the training phase. Once operative technique has been mastered, a success rate of greater than 90% should be achieved before inclusion of the animals in further scientific studies. As the AOLT animals recover body weight faster and have less evidence of liver injury up to 4 weeks post-transplantation, one is led to the conclusion that AOLT in the rat may be a better model for use in all liver transplantation research. PMID- 8404989 TI - Role of nonparenchymal liver cells in guinea pig to rat hepatic xenotransplantation. AB - This work was undertaken to investigate the role of nonparenchymal liver cells in a discordant model of hepatic xenografting. Three experimental groups were established: in group 1 guinea pig to Lew rat liver xenotransplantations were performed; in group 2 both donor and recipient were treated with dextran sulfate, a known inhibitor of the reticuloendothelial system phagocytic function; in group 3 both donor and recipient were injected with muramyl dipeptide, a synthetic immunomodulator stimulating the monocyte/macrophage axis. Survival time was assessed and xenoantibody titers 30 min before and after the intervention, Kupffer cell activity 30 min after transplantation, histology and immunoglobulin and complement deposits of the grafted liver were evaluated too. Survival time of the xenografted rats in group 1 was 6.4 +/- 0.31 h. Blockade of Kupffer cells by dextran sulfate administration significantly (p < 0.001) depressed the survival time (2.9 +/- 0.31 h) of the grafted rats, while a significant increase (p < 0.001) was observed in the muramyl dipeptide-treated group (9.3 +/- 0.52 h). A significant reduction of xenoantibody titers 30 min after intervention was observed in the muramyl dipeptide group while no reduction was observed in the dextran group. Thirty minutes after xenotransplantation sheep red blood cell 51Cr uptake was significantly depressed by dextran sulfate treatment while muramyl dipeptide administration restored the Kupffer cell activity. Histological changes worsened after dextran administration in comparison with the other groups. Immunoglobulins and complement deposits were diminished by dextran administration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8404990 TI - Increase in intraepithelial lymphocytes as an early marker of rejection in a fully allogeneic rat small bowel transplantation model. AB - In a model of fully allogeneic heterotopic rat small bowel transplantation, the changes in intraepithelial lymphocyte (IEL) number and subpopulations were analysed. During early phases of rejection (5th postoperative day) a 4-fold increase in the number of IEL was observed when compared with native small bowel (4.05 vs. 15.84 IEL/100 epithelial cells). When cyclosporine was given, counts were still as high as 11.4 and 12.58 on the 5th and 10th postoperative days, respectively. The percentage of CD8+ IEL, constituting a major population (84%) in the untreated small bowel, was significantly decreased (46.4%) during early phases of rejection. At that time, the majority of intraepithelial mononuclear cells were both CD8- and CD4-. In cyclosporine-treated animals, this was not observed until the 10th postoperative day. Some 23% of IEL in untreated animals expressed MHC class I antigens of the host; 17.2% (5th postoperative day) and 19.8% (10th postoperative day) did so in the cyclosporine-treated animals. Transmission electron microscopy revealed lymphocytes that bore cytoplasmic buds and pseudopods protruding between the enterocytes. There was no morphological difference between the IEL of rejected allografts and native small bowel. Due to the unspecific histologic changes associated with rejection, interpretation of histopathologic findings in mucosa biopsies of the allograft can be rather troublesome. An increase in the number of IEL is therefore a welcome additional marker of rejection. PMID- 8404991 TI - Regeneration of sympathetic activities in small bowel transplants. AB - In order to study the activity of noradrenergic nerve fibres along mesenteric arteries in small-intestinal grafts, the entire jejunoileum was transplanted heterotopically in an isogeneic rat model (group I, n = 12). To assess their influence on graft motility, 9 cm of jejunum were transplanted in an orthotopic position and three bipolar electrodes sutured to the seromuscular layer of the graft (group II, n = 10). Fasting motility was recorded up to postoperative day 42. Animals of group I were sacrificed from day 7 on at weekly intervals and mesenteries were analysed histochemically by fluorescence microscopy. After the 1st and 2nd week, grafts were found to be completely depleted of noradrenaline. At the end of the 3rd week, fluorescence became detectable along graft mesenteric arteries and showed normal intensity from the end of week 4. Migrating myoelectric complexes (MMCs) were present in all animals of group II. The variability of the MMC period (mean 12.6 min; SD 6.2 min) expressed as variation coefficient (median 36.5; 14.6-74) did not change during the observation period. From these findings it is concluded that there are no extrinsic noradrenergic activities in rat small-intestinal transplants for the first 3 postoperative weeks but they do recover there after. Their influence on graft function remains unclear: MMC periodicity, however, was no influenced. PMID- 8404992 TI - The reduced canine pancreas to study the effects of intra-operative radiotherapy. AB - A canine model is described to study the tolerance of the pancreas to intra operative radiotherapy (IORT). The canine pancreas is a horseshoe-shaped organ. To create a homogeneous delivery of IORT to the whole pancreas surgical manipulation is necessary which may induce pancreatitis. A resection of the left and right lobes of the pancreas facilitates the delivery of IORT, reduces the risk of pancreatitis and will demonstrate, eventually, minimal functional changes in the exocrine and endocrine pancreas at an earlier stage. Sixteen beagles were used. Investigations before and after the reduction procedure were intravenous glucose tolerance tests, serum insulin levels, faecal fat excretion, blood chemistry tests and body weight. Eight weeks after the pancreas reduction 15 dogs underwent an IORT procedure in which 25, 30 or 35 Gy IORT was delivered to the pancreatic remnant. We conclude that the pancreas reduction technique used to study the effects of IORT to the canine pancreas is feasible without mortality or morbidity. Endocrine and exocrine pancreatic function remained normal with a minimal follow-up of 3 months. PMID- 8404993 TI - PGI2 aerosol versus nitric oxide for selective pulmonary vasodilation in hypoxic pulmonary vasoconstriction. AB - Intravenous prostacyclin (PGI2) is a potent pulmonary vasodilator in pulmonary hypertension. However, dose-dependent systemic vasodilation, an increase in intrapulmonary shunt and hypoxemia limit its clinical application. Recently, inhaled nitric oxide (NO) has been reported to elicit selective pulmonary vasodilation, but its clinical use is restricted by its potential toxicity; furthermore, the feasibility of NO application in clinical practice seems difficult. Therefore, we investigated the effects of PGI2 aerosol on pulmonary and systemic circulation and compared the hemodynamic effects to those of inhaled NO. In 6 dogs, ventilation with a hypoxic gas mixture (FiO2 0.09-0.11) increased pulmonary vascular resistance (PVR) by 196% (HPV). Aerosolization of a PGI2 solution at a concentration of 430 ng/ml reduced hypoxia-induced increase of pulmonary artery pressure by 48% and PVR by 52% within 6-10 min without systemic vasodilation. The administered dose of PGI2 was 0.87 +/- 0.26 ng/kg/min. In 2 dogs, doubling the PGI2 concentration (860 ng/ml) did not enhance the vasodilatory effect. After termination of PGI2 inhalation, HPV was restored within 10-15 min. Inhaled NO (50 ppm) decreased the HPV-induced increase in PAP by 76% and in PVR by 73% within 5-10 min. Clinically relevant systemic vasodilation was not observed. It is concluded that inhalation of aerosolized PGI2 leads to selective pulmonary vasodilation in hypoxia-induced pulmonary hypertension. Aerosolized PGI2 at a concentration of 430 ng/ml was less potent than NO (50 ppm). However, due to the lack of known toxicity and its uncomplicated mode of application, inhaled PGI2 may be one alternative to inhaled NO in the treatment of acute pulmonary hypertension. PMID- 8404994 TI - Inhaled nitric oxide--from pollutant to patent. PMID- 8404995 TI - Dose-response effects of lactate infusions on gluconeogenesis from lactate in normal man. AB - Lactate is the predominant gluconeogenic precursor in man. To determine the dose response relationships between plasma lactate concentration and rates of lactate incorporation in plasma glucose (lactate gluconeogenesis, LGN), we infused 17 normal volunteers with sodium lactate for 180 min at rates ranging from 6 to 40 mumol kg-1 min-1 and measured [U-14C]lactate incorporation into plasma glucose, as well as rates of lactate and glucose appearance in plasma. With the highest lactate infusions, plasma lactate increased up to 7 mM (compared to 1.1 +/- 0.13 mM during control sodium bicarbonate infusions, n = 10) and LGN averaged 4.73 +/- 0.23 mumol kg-1 min-1 (compared to 1.57 +/- 0.26 mumol kg-1 min-1 in bicarbonate control experiments, P < 0.001). The data relating plasma lactate concentration to LGN best fit a sigmoidal curve which plateaued at plasma lactate concentrations of approximately 6 mM and yielded an ED50 of 2.04 +/- 0.20 (SD) mM and a Vmax (6.25 +/- 1.2) (SD) (mumol kg-1 min-1). The sum of the basal rate of lactate appearance and the rate of lactate infusion was not significantly different from the overall rates of lactate appearance during the lactate infusions (35.8 +/- 2.2 vs. 34.8 +/- 2.9 mumol kg-1 min-1, P = 0.23). Thus, our results support the view that infusion of exogenous lactate does not suppress endogenous lactate appearance in plasma. PMID- 8404996 TI - Affinity of anti-GP41 antibody in patients infected with human immunodeficiency virus type 1. AB - Anti-gp41 antibody affinity was investigated prospectively in 25 patients with asymptomatic and symptomatic HIV-1 infection for a period of 9-42 months. Major differences in the processes of immune response maturation towards gp41 were observed among individual subjects, however, antibody affinity increased with time in all examined persons including patients with AIDS. Anti-gp41 affinity values were found to reflect both the duration and the clinical stage of HIV infection. PMID- 8404997 TI - Decreased ability of high density lipoproteins to transfer cholesterol esters in non-insulin-dependent diabetes mellitus. AB - Esterified cholesterol transfer (ECT) from high density lipoproteins (HDL) to very low (VLDL) and low density lipoproteins (LDL) may be abnormal in situations at high risk for atherosclerosis. It has been shown to increase in insulin dependent diabetes and to decrease in non-insulin-dependent diabetes (NIDD). Since the net transfer of esterified cholesterol (EC) results from a bidirectional exchange between HDL and VLDL/LDL, we developed a transfer assay specifically designed to measure the unidirectional transfer of EC from HDL to lipid emulsions according to first-order kinetics. Our results show that in NIDD the rate constant of HDL-dependent ECT is decreased by 30% by comparison with control subjects. Analysis of HDL composition revealed that, in both groups, HDL dependent ECT was positively correlated with the free cholesterol/phospholipid ratio (r = 0.94; P < 0.001) and negatively correlated with the triglyceride/EC ratio (r = -0.85; P < 0.001). It is concluded that, besides the known defect of acceptor lipoproteins, the abnormality of ECT in NIDD is also caused by a decreased ability of HDL to act as an EC donor, presumably because of a change in composition. In addition, our work shows that the amount of EC lost by HDL during the reaction transfer is counterbalanced by a reciprocal equimolar transfer of triglycerides. PMID- 8404998 TI - The effects of the menopause on insulin sensitivity, secretion and elimination in non-obese, healthy women. AB - We have carried out intravenous glucose tolerance tests with measurement of plasma glucose, insulin and C-peptide concentrations on 66 premenopausal and 92 postmenopausal non-obese caucasian women. After adjustment for the effects of a number of possible confounding variables, including age and body mass index, there was little difference between pre and postmenopausal women in glucose and insulin concentrations either fasting or in response to intravenous glucose. Mathematical modelling analysis of the resultant plasma concentration profiles was used to obtain measures of insulin sensitivity, secretion and elimination, and non-insulin dependent glucose disposal. We found reciprocal differences in mean insulin sensitivity (increased by 50%) and non-insulin dependent glucose disposal (decreased by 30%). Plasma C-peptide response and pancreatic insulin secretion were markedly lower in the postmenopausal group (-35% and -51% respectively). However, the rate constant for insulin elimination was also lower in these women. As a result, intravenous glucose tolerance test plasma insulin concentrations were not significantly different between the two groups. We conclude that, despite the occurrence of little or no variation in plasma glucose and insulin concentrations, the menopause is associated with significant changes in insulin metabolism. PMID- 8404999 TI - Monocyte adherence to endothelial cells in patients with atherosclerosis: relationships with risk factors. AB - Monocyte adhesion to endothelium appearing determinant in atherosclerosis, the possibility that circulating monocytes have an increase of their adherence on endothelial cells were investigated in patients with atherosclerosis. The adherence of circulating monocytes on endothelial cell monolayers was determined in 26 patients with atherosclerosis (age 59 +/- 4), and 25 healthy individuals (age 55 +/- 4). No difference of monocyte adherence was observed between the two groups (18.8 +/- 13.8% vs 19.2 +/- 13.4%), or following atherosclerosis severity. However, monocyte adherence appeared positively correlated to smoking habits (r = 0.34, P < 0.02) and fibrinogen level (r = 0.31, P < 0.03), and negatively to the degree of plasmatic LDL oxidation (r = -0.28, P < 0.05). These results suggest that the adherence of monocytes to endothelial cells is not increased in atherosclerosis, but enhanced by risk factors. A weak plasmatic LDL oxidation could inhibit monocyte adhesion. PMID- 8405000 TI - Insulin induced hypoglycaemia and sugar transport across the brush border and basolateral membranes of rat jejunal enterocytes. AB - Acute hypoglycaemia enhances intestinal sugar uptake but the mechanisms involved are unknown. Results from the present study show increased galactose movement across the brush border and basolateral membranes of isolated upper, but not mid villus, jejunal enterocytes 45 min after intravenous administration of insulin to rats at a level which reduced by half plasma glucose concentration. Incubation of upper villus cells from uninjected animals with insulin (100 mU ml-1) for 40 min was without effect on brush border or basolateral sugar transport. Insulin treatment of rats did not affect glucose uptake by brush border vesicles prepared from upper villus cells when the process was driven by an inwardly directed 100 mM sodium thiocyanate gradient. In contrast, glucose uptake using a 100 mM inwardly directed sodium chloride gradient was reduced by 49% following hypoglycaemia. It is concluded that the enhanced sugar uptake following insulin hypoglycaemia involves both brush border and basolateral membranes of only the most mature villus cells at the villus tip. Upregulation of Na(+)-sugar cotransport at the brush border is best explained by an increased electrochemical driving force for Na(+)-sugar cotransport rather than increased numbers of transporters. The transport response is not due to a direct effect of insulin on the enterocyte and the possible systemic factors involved are discussed. PMID- 8405001 TI - Relationship of alcohol consumption to changes in HDL-subfractions. AB - Epidemiological studies have consistently shown an apparent protective effect of moderate alcohol consumption on coronary artery disease (CAD). This has been considered to be due to the rise in the high-density cholesterol lipoprotein (HDL cholesterol). Since the response of the HDL-subfractions to moderate or heavy dose of alcohol is less clear, we now compared the high-density lipoprotein cholesterol status between groups consuming different amounts of alcohol. In this population-based survey serum total high-density lipoprotein cholesterol and its HDL2 and HDL3 subfractions were blindly compared between 264 consecutive middle aged men (37 teetotallers, 137 moderate drinkers, 90 heavy drinkers) participating in a voluntary health screening and 104 male alcoholics. Alcohol consumption correlated significantly (P < 0.001) with total HDL-cholesterol, HDL2, HDL3 when all subjects (n = 368) were included but the correlation disappeared when alcoholics were excluded (n = 264). In comparison with teetotallers, alcoholics had significantly higher total HDL-cholesterol, HDL2 and HDL3 values (P < 0.001). Moderate or heavy intake of alcohol had no effect on HDL2 but increased the HDL3-fraction. If the protective effect of moderate alcohol consumption is mediated by high-density lipoprotein, it may not be accounted for by changes in the HDL2-fraction. The observed increases in the concentration of the HDL3-fraction, however, suggest that this subfraction may not be inert with respect to coronary disease and could possibly have a role in the protective effect. PMID- 8405002 TI - Disparate effects of a triglyceride lowering diet and of bezafibrate on the HDL system: a study in patients with hypertriglyceridaemia and low HDL-cholesterol levels. AB - The response of plasma triglyceride and of the high density lipoprotein (HDL) system to a triglyceride lowering diet and to a bezafibrate was compared in a group of 24 human subjects with mild to moderate hypertriglyceridaemia and low HDL-cholesterol levels. Post-heparin plasma lipoprotein (LPL) and hepatic (HL) lipase activities were determined before the initiation of the study and at the end of the diet (12 weeks) and bezafibrate (12 weeks) periods. HDL structure and composition were determined on a zonal centrifugation system at the end of the diet and bezafibrate periods. Diet caused a 9-20% reduction of plasma triglycerides but there was no change in LPL, HL or HDL levels. The individual responses between plasma triglycerides and HDL cholesterol levels however were highly correlated (r = 0.60 to 0.78). Combined diet plus bezafibrate therapy caused a further 38% decrease of plasma triglycerides with a 28% increase of HDL cholesterol and a 47% increase of LPL activity. HDL3 density decreased and the contribution of cholesteryl ester to the HDL mass was increased while the contribution of proteins and triglycerides decreased. In the majority of the subjects (61%) HDL cholesterol levels increased by 20% or more and in these subjects the change of HDL was highly correlated with the change of the LPL/HL ratio (r = 0.74) but not with the change of the plasma triglyceride levels. In the remaining subjects (39%), HDL cholesterol levels remained unchanged or increased by less than 20%. The change of HDL cholesterol levels in these subjects was unrelated to the decrease of plasma triglycerides or the change of post-heparin lipase activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405003 TI - Time-course and dose-response of nitric oxide inhalation for systemic oxygenation and pulmonary hypertension in patients with adult respiratory distress syndrome. AB - Inhalation of nitric oxide (NO), an endogenous vasodilator, was recently described to reduce pulmonary vascular resistance, and to improve arterial oxygenation by selective vasodilation of ventilated areas in patients with adult respiratory distress syndrome (ARDS). We describe the time-course and dose response of initial short-term NO inhalation in 12 patients with ARDS. Enhanced oxygenation was achieved within 1-2 min after starting NO inhalation; after inhalation, baseline conditions were re-achieved within 5-8 min. Effective doses for improvement of oxygenation [baseline: PaO2 = 10.2 +/- 2.5 KPa (76.4 +/- 18.7 mmHg)] were low: ED50 was about 100 ppb--a concentration similar to the atmosphere. NO doses of more than 10 ppm [10 ppm NO: PaO2 = 17.3 +/- 3.3 KPa (129.4 +/- 25.1 mmHg)] re-worsen the arterial oxygenation. The ED50 for reduction of mean pulmonary artery pressure was 2-3 ppm. This indicates that inhalation of NO for improvement of oxygenation in severe ARDS should be performed using lower doses, with lower risk of toxic side effects. PMID- 8405004 TI - Physiological modelling of renal drug clearance. AB - A physiological model of renal drug clearance is presented with the aim of establishing a basis for adjusting drug dosing regimens in renal insufficiency. In agreement with the morphology of blood supply to the nephron, the model assumes serial arrangement of the processes involved in drug excretion. Fractional extraction by filtration in the glomeruli is defined in terms of the product of the unbound fraction of the drug, the filtration fraction being responsible for the limited extraction efficiency of this process. For a description of the limitations of the tubular secretory process by plasma flow through peritubular capillaries, the parallel tube model is utilized. The assumption of direct proportionality between the transport maximum of the secretory process and filtrate flow in the tubules permits a quantitative comparison of the intrinsic tubular secretion clearance and the effectiveness of the filtration process. Provided that the secretory mechanism is highly effective, renal clearance becomes dependent only on kidney plasma flow and the fraction of drug not reabsorbed in the tubules. Tubular reabsorption results only in a proportional decrease in renal clearance. The model predicts proportionality of renal drug clearance to GFR, which as a rule is used for dosage adjustment of drugs in renal insufficiency, only for compounds exclusively excreted by filtration. Compounds also excreted by tubular secretion in general exhibit a curvilinear relationship. The curvature is less pronounced as an increasing fraction of the drug is protein bound in blood. Therefore, for dosage adjustment of drugs secreted in the tubules and highly bound in blood, proportionality between renal clearance and GFR can serve as a reasonable approximation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405005 TI - Quantitative analysis of drug handling by the kidney using a physiological model of renal drug clearance. AB - Published data on the renal clearance of creatinine, p-aminohippuric acid (PAH) and kanamycin in relation to glomerular filtration rate (GFR) in patients with various renal diseases were analysed by a physiological model of renal clearance. Fitting of the data by the general linear equation representing the model proposed by Levy [10] resulted in insignificant intercepts with the ordinate, indicating the unsuitability of the model for the detection of tubular secretory activity. Use of this model also did not lead to significant improvement in goodness of fit compared to simple proportionality of renal clearance and GFR. On the other hand, parameter estimates of the physiological model obtained from the data by nonlinear regression analysis revealed statistically significant tubular secretion both of PAH and creatinine. The much lower tubular secretory activity estimated from the kanamycin data did not reach statistical significance. For compounds exhibiting statistically significant tubular secretion, use of the physiologically based relationship between renal clearance and GFR significantly improved the goodness of fit to the data as compared to simple proportionality of both variables. It is concluded that analysis of the relationship between renal clearance of drugs and GFR using the physiological model of renal clearance can contribute to our knowledge of drug handling by the kidney, and may facilitate drug classification according to total extraction by this organ. PMID- 8405006 TI - The effect of midazolam on transient insomnia. AB - We have assessed the effect of midazolam on sleep in a model of transient insomnia in healthy adults using polysomnographic recordings. The subjects were randomly assigned to one of three treatment groups (placebo or midazolam 7.5 or 15 mg) and spent a single night in the sleep laboratory. Midazolam or placebo were given double-blind immediately before turning off the lights. Midazolam 15 mg was effective in inducing sleep, while 7.5 mg and 15 mg produced improvement in the maintenance of sleep. Subjectively, sleep latency and the number of awakenings were reduced dose-dependently. Midazolam did not impair psychomotor performance on the morning after administration. PMID- 8405007 TI - Memory and psychomotor effects of oxcarbazepine in healthy human volunteers. AB - Cognitive and psychomotor impairments can be unwanted adverse effects of antiepileptic drugs. The present double-blind, cross-over study with healthy volunteers was designed to assess the effects of two doses of oxcarbazepine (OXCZ) (150 mg b.i.d.; 300 mg b.i.d.) and a placebo, each given over a two week period. Twelve subjects completed a battery of tests before and 4 h after morning doses on days 1, 8 and 15. Results of objective tests indicated that OXCZ improved performance on a focussed attention task and increased manual writing speed. Subjective ratings showed OXCZ increased feelings of altertness, clear headedness and quickwittedness. OXCZ had no effect on the range of long-term memory processes assessed in this study. It is concluded that at the doses employed, OXCZ has a slightly stimulant effect on some aspects of psychomotor functioning. PMID- 8405008 TI - Combined therapy with probucol and pravastatin in hypercholesterolaemia. One year follow-up study. AB - The effect of co-administration of low doses of pravastatin to hypercholesterolaemic patients already receiving long-term probucol treatment (mean 500-1,000 mg/day for 350 days) were investigated. Pravastatin 5 mg/day (Group 1; 12 m, 13 f; mean age 59.1 y) or 10 mg/day (Group 2; 8 m, 11 f; mean age 60.8 y) was administered, and blood was taken after 0, 3, 6, and 12 months. Both groups showed a significant reduction in serum total cholesterol (TC), phospholipid (PL), low density lipoprotein-cholesterol (LDL-C), LDL-triglyceride (TG), LDL-PL, apolipoprotein (apo) B, and apo E after the combined therapy. These levels were reduced more in Group 2 than in Group 1 subjects. In Group 2, significant falls in serum TG and apo CII were also observed. The changes in TC, PL, LDL-C, apo B, apo CII and apo E were dependent upon the dose of pravastatin, as assessed by two-way analysis of variance. Serum high density lipoprotein (HDL)3-C, apo AI and apo AII were slightly but significantly increased in both groups after 12 months of combined therapy, but the increase was not sufficient to reverse the probucol-induced lowering of the HDL level. We conclude that combined therapy resulted in a significant reduction in atherogenic lipoproteins and apolipoproteins, and an increasing dose of pravastatin (5 mg to 10 mg daily) made the lipid lowering effect more prominent. The reduction in serum HDL-C due to long-term probucol administration was not reversed by the addition of pravastatin. PMID- 8405009 TI - Inhibitory effects of inhaled flunisolide on inflammatory functions of alveolar macrophages. AB - We have studied 15 patients with slight or moderate bronchial obstruction, all of whom were being treated by inhalation of the beta-mimetic fenoterol 4 x 400 micrograms/day, and 7 of whom were also receiving inhaled flunisolide 2 x 500 micrograms/day. The therapy had been given for longer than 1 month in each case. Bronchoscopy and bronchoalveolar lavage (BAL) was done for diagnosis or follow up of bronchial diseases. None of the patients showed signs of any interstitial lung disease. Conditioned culture supernatants were produced by cultivating alveolar macrophages (AM) for 24 h using standard conditions. To detect all the biological effects both of IL-1 alpha and IL-1 beta in the culture supernatants a modification of the standard mouse IL-1 thymocyte bioassay was used. The TNF concentration in culture supernatants was measured by ELISA. Free oxygen radical release by alveolar macrophages was determined by the detection of chemiluminescence. Both IL-1 and TNF production were significantly lower in patients receiving fenoterol plus flunisolide than in patients on fenoterol alone. In contrast, no difference could be observed in the release of free oxygen radicals from alveolar macrophages. Thus, for the first time an ex vivo study has revealed an interrelation between inhaled glucocorticoid therapy and inhibition of important mediators of inflammatory processes in the lower respiratory tract. PMID- 8405010 TI - Effect of ipratropium on nasal reactivity to histamine and eosinophil influx in perennial allergic rhinitis. AB - In a double-blind, placebo-controlled study nasal saline and histamine provocation tests were performed in patients with perennial allergic rhinitis in order to assess changes in eosinophil influx and non-specific nasal reactivity after 8 days of treatment with ipratropium bromide. A "nasal pool" method was used to trace changes in protein level and eosinophil influx into nasal secretions. Treatment with ipratropium 80 mg q.i.d. caused a significant decrease in the albumin and total protein level in saline washings and induced a five-fold increase in eosinophils as compared to the placebo treatment. The nasal mucosal response to histamine, assessed as the number of sneezes and protein level, was more responsive to ipratropium treatment than the mucosa from placebo-treated subjects. Since eosinophil numbers were correlated with an increase in the vascular and sneezing responses, it appears that ipratropium potentiates inflammatory mechanisms when used in subjects with an allergy in the nasal mucosa. PMID- 8405011 TI - Duration of caffeine abstention influences the acute blood pressure responses to caffeine in elderly normotensives. AB - The effect of an acute oral caffeine load (250 mg capsule) and matching placebo on blood pressure and pulse rate were studied after 48 h caffeine abstention in 8 elderly, normotensive regular caffeine users. The caffeine loading phase was repeated after only 12 h abstention. Following 48 h abstention, supine systolic and diastolic blood pressure were higher for the 120 min study period after the acute caffeine load than placebo (12.1 mm Hg, 95% C.I. 4.3-19.9 mm Hg; P = 0.008 and 7.4 mm Hg, 3.6-11.2 mm Hg; P < 0.001 respectively). Similar differences were seen in standing blood pressure, though pulse rate was unchanged throughout. The pressor response to the acute caffeine load was significantly greater after a 48 h than a 12 hour caffeine abstention period, for supine and standing systolic and diastolic blood pressure. The changes in plasma caffeine levels after acute loading were similar after the 2 different abstention periods. Caffeine ingestion after 48 h abstention has an acute pressor effect in normotensive elderly subjects which is abolished if the abstention period is reduced to 12 h. Acute caffeine ingestion is unlikely to have a significant pressor effect in elderly normotensive subjects who are regular caffeine users as the normal period of caffeine abstention (i.e. overnight) is too short to abolish tolerance. PMID- 8405012 TI - Plasma concentrations of bupivacaine after wound infiltration of an 0.5% solution after inguinal herniorrhaphy: a preliminary study. AB - After routine inguinal herniorrhaphy we gave 12 patients a wound infiltration regimen of bolus doses of 20 ml of 0.5% bupivacaine via a catheter within the wound and rectally administered indomethacin (100 mg). Peak venous plasma bupivacaine concentrations ranged from 0.07 mg.l-1 to 1.14 mg.l-1 (mean (SD) 0.47 (0.33) mg.l-1), and occurred at between 0.25 and 2 h after the first dose. Plasma concentrations were well below the toxic threshold of 4 mg.l-1 and there was no accumulation. The regimen provided satisfactory analgesia. There were no wound infections nor signs of toxicity. PMID- 8405013 TI - Development and preliminary application of a simple assay of S-mephenytoin 4 hydroxylase activity in human liver microsomes. AB - We have developed a simple HPLC assay to measure the activity of S-mephenytoin 4 hydroxylase in human liver microsomes, and have assessed its practical applicability by determining the kinetic parameters of the enzyme in 10 different human liver samples. The recovery of 4-hydroxymephenytoin and phenobarbital (the internal standard) after the precipitation of microsomal protein was almost complete, and the coefficients of variation for the intra- and interassay measurement of S-mephenytoin 4-hydroxylase activity were < 6.4 and 8.0%, respectively. Eadie-Hofstee plots for the formation of 4-hydroxymephenytoin gave a straight line for all of the 10 samples studied. There was large interindividual variability in the kinetic parameters estimated: 4.6- (36 to 166 microM), 11.8- (0.9 to 10.6 nmole/mg protein/h) and 30.1- times (0.10 to 3.01 microliters/mg protein/min) for Km, Vmax and Vmax/Km, respectively. The mean (+/- SD) Km, Vmax and Vmax/Km were 72.4 +/- 40.4 microM, 4.23 +/- 2.88 nmole/mg protein/h and 1.33 +/- 1.02 microliters/mg protein/min, respectively. Thus, the assay was sufficiently accurate and reproducible to permit estimation of the kinetic parameters of S-mephenytoin 4-hydroxylase in human liver microsomes, and it appears to be applicable to an in vitro study of the possible involvement of S mephenytoin-type oxidation polymorphism in drug metabolism. PMID- 8405014 TI - Pharmacokinetic evaluation of oral 17 beta-oestradiol and two different fat soluble analogues in ovariectomized women. AB - A randomised, single-blind comparative study was carried out in 9 ovariectomized women to evaluate the kinetics of single doses of three different steroid combinations: 0.150 mg desogestrel + 2.0 mg micronized 17 beta-oestradiol, 0.150 mg desogestrel + 0.500 mg 17 beta-oestradiol cyclo-octyl acetate and 0.150 mg desogestrel + 1.0 mg 17 beta-oestradiol decanoate. Serum levels of 17 beta oestradiol and oestrone were measured, as well as the excretion of 17 beta oestradiol and its metabolites (oestrone and oestriol) in urine. In relation to the doses given, higher peak serum concentrations of 17 beta-oestradiol were obtained after the two fat soluble analogues, while the AUCs were similar to that after micronised 17 beta-oestradiol. However, there was more extensive conversion of the micronised 17 beta-oestradiol preparation into oestrone compared to 17 beta-oestradiol cyclo-octyl acetate and 17 beta-oestradiol decanoate. The oestrone/17 beta-oestradiol serum concentration ratio was approximately 2.6 before tablet intake and remained essentially unchanged after intake of 17 beta oestradiol cyclo-octyl acetate and 17 beta-oestradiol decanoate. After micronized 17 beta-oestradiol however, there was a 2-3-fold increase in the ratio at Cmax and slower elimination of 17 beta-oestradiol from plasma, which may be due to the fact that high serum oestrone levels may serve as a reservoir, since both a metabolite and also a precursor of 17 beta-oestradiol. The urinary excretion of 17 beta-oestradiol, oestrone and oestriol was highest after oral administration of micronized 17 beta-oestradiol compared to 17 beta-oestradiol cyclo-octyl acetate and 17 beta-oestradiol decanoate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405015 TI - Pharmacokinetic comparison of leucovorin and levoleucovorin. AB - The pharmacokinetic values of d,l-leucovorin and l-leucovorin were compared in eight healthy volunteers following oral administration of 25 mg d,l-leucovorin and 12.5 mg l-leucovorin. Serum levels of l-5-formyltetrahydrofolate, l-5 methyltetrahydrofolate, and total reduced folates were measured by an established microbiological method. Pharmacokinetic data for both preparations were consistent with those previously reported for d,l-leucovorin, with essentially complete first pass metabolism to l-5-methyltetrahydrofolate, the active metabolite. No differences were found between the two preparations in serum concentrations of active folate fractions, AUC, or Cmax, or in clearance and volume of distribution estimates. These data suggest that after administration of 25 mg of d,l-leucovorin, the d-diastereoisomer has no significant effect on the standard pharmacokinetic measurements of the active l-folates. PMID- 8405016 TI - Pharmacokinetics of pantoprazole following single intravenous and oral administration to healthy male subjects. AB - The plasma pharmacokinetics of pantoprazole have been investigated following single intravenous infusion and single oral administration at a dose of 40 mg to 12 healthy male subjects in a randomised cross-over study. Both treatments were generally well tolerated and no relevant compound-related adverse events were noted. The plasma pharmacokinetics of pantoprazole following intravenous infusion in this group of subjects were characterised by a total plasma clearance of 0.13 l.h-1 x kg-1 and apparent terminal elimination half-life 1.9 h. The apparent volume of distribution estimated at steady state (0.17 l.kg-1) was compatible with the localization of a major fraction of the compound in extracellular water. Following oral administration as an enteric-coated tablet formulation, a variable onset of absorption was followed by rapid attainment of maximum plasma concentrations of pantoprazole. Pantoprazole was well absorbed following oral administration; the absolute systemic bioavailability of the compound was estimated as 77% (95% CI, 67 to 89%). PMID- 8405017 TI - Lack of a pharmacokinetic interaction between carvedilol and digitoxin or phenprocoumon. AB - The possibility of a pharmacokinetic interaction between carvedilol and digitoxin (Study I) or phenprocoumon (Study II) has been evaluated in groups of 12 healthy volunteers. The bioavailability (Cmax, tmax, AUC) of digitoxin and phenprocoumon were assessed after a single dose, given once alone and once on day 6 of treatment with carvedilol 25 mg o.d. Cmax, tmax, AUC and Ut of carvedilol and desmethylcarvedilol were also investigated after the fifth dose of carvedilol and after the sixth dose given concomitantly with digitoxin or phenprocoumon. In Study I, the 95% confidence intervals of the ratio test versus the reference findings were; digitoxin Cmax 0.80-1.20, tmax 0.56-1.14, AUC 0.97-1.33, and for carvedilol Cmax 0.81-1.22; tmax 0.66-1.23; AUC 0.91-1.17. Formation of the active metabolite desmethylcarvedilol and the urinary recovery of carvedilol and desmethylcarvedilol were not influenced by digitoxin. In Study II Cmax and AUC of phenprocoumon were not changed after carvedilol. Cmax of carvedilol was decreased after phenprocoumon. The kinetic parameters of phenprocoumon were Cmax 0.80-1.05, tmax 0.47-2.00, AUC 0.78-1.05, and for carvedilol Cmax 0.59-1.06, tmax 0.71-1.73; AUC 0.80-1.08, respectively. The plasma levels of desmethylcarvedilol and the urinary recovery of carvedilol and desmethylcarvedilol were not influenced by phenprocoumon. The blood pressure and heart rate after carvedilol alone were not affected by concomitant administration of digitoxin or phenprocoumon. PMID- 8405018 TI - The analgesic efficacy of diclofenac dispersible and ibuprofen in postoperative pain after dental extraction. AB - We have compared single oral doses of drinkable diclofenac dispersible (50 mg) with ibuprofen (400 mg) and placebo in a randomized, double-blind, parallel-group trial in 127 adults complaining of at least moderately severe pain after removal of an impacted third molar. Within 40 min both diclofenac and ibuprofen produced significant pain relief that persisted for 6 h. There were no differences between diclofenac and ibuprofen in analgesic efficacy. PMID- 8405019 TI - Metformin-associated lactic acidosis in Sweden 1977-1991. AB - Since the withdrawal of phenformin in 1978, the use of metformin has increased from 13,500 to 22,000 patient years/year. During the period 1977-91 a total of 18 cases of metformin-associated acidosis was reported, of which 16 had lactic acidosis. The incidence of reported acidosis and lactic acidosis decreased from 1.50 cases per 10,000 patient years in 1977-81 to 0.24 cases per 10,000 patient years 1987-91, probably due to lower doses doses and reduced usage in the very old. All the reports described patients with several other concomitant diseases, mainly cardiovascular and renal, when the acidosis was diagnosed. It is important continuously to re-evaluate metformin therapy and to stop treatment at the onset of impaired renal or cardiovascular function. PMID- 8405020 TI - Frequency of S-mephenytoin hydroxylation deficiency in 373 Spanish subjects compared to other Caucasian populations. AB - We have investigated the prevalence of poor metabolisers (PM) of S-mephenytoin in 373 unrelated, healthy Spanish Caucasian subjects, based on the enantiomeric S/R mephenytoin ratio in urine collected 0-8 h and 24-32 h after intake of the racemic drug. Five of the subjects were PM (1.34%, 95% confidence interval 0.18 2.59%), a prevalence lower than in 6 other Caucasian populations, but only significantly lower than in studies in France and Switzerland (P < 0.01). We suggest that this difference might be due to the use of different phenotyping procedures. PMID- 8405021 TI - Parkinson's disease study. PMID- 8405022 TI - Is it dexfenfluramine or weight loss that reduces blood pressure and noradrenergic activity in obese patients? PMID- 8405023 TI - Clinical and statistical issues in therapeutic equivalence trials. AB - Absolute proof of efficacy can only be given by placebo controlled trials. It is, however, important to classify a drug within the spectrum of existing therapeutic alternatives and, where effective treatment is available, it may be imperative due to ethical considerations to demonstrate that one drug is as effective as another. The issue of therapeutic equivalence trials is discussed along the lines of the important items which should be defined in the protocol: a) the target parameter, which is the primary endpoint of the trial, b) the reference drug, which should be selected with respect to efficacy (superior to others), and safety (largest amount of data), c) the acceptance range, which depends on the primary endpoint, and its implication for the clinical endpoints of morbidity and mortality (the conventional acceptance range for bioequivalence trials does not apply), and d) the statistical procedures, which must take into consideration the unsuitability of the conventional power approach for confirming equivalence. In an equivalence trial, compared to those that are placebo-controlled, the proof that one drug is as effective as another relies much more upon the quality of conduct of the study according to Good Clinical Practice. PMID- 8405024 TI - Motives and perception of healthy volunteers who participate in experiments. AB - Healthy volunteers who participated in an experiment were asked to complete a questionnaire about their experiences. Special attention was paid to their motives and the informed consent procedure. The motives of healthy volunteers for participating in a study differed according to age. Young volunteers mostly participated because of the financial compensation, while older volunteers mainly participated to get a medical check-up, which was part of the selection procedure, or because of the benefit that other people might obtain from the results of the experiment. In most experiments the volunteers were content with the information given about the experiment, which was provided orally and in a hand-out. The information procedure was crucial for the experiment, because optimum information was of the utmost importance in keeping the volunteers motivated during the experiment. Another conclusion was that the researcher should not be afraid that too much information was given. Adequate information was also important in recruiting new volunteers, because they mostly become interested because of information obtained from their predecessors. A personal approach towards the volunteers during the experiment was appreciated by them and was an important help in keeping the volunteers motivated during the experiment. PMID- 8405025 TI - Effect of chronic treatment with enalapril on glucose tolerance and serum insulin in non-insulin-resistant Japanese patients with essential hypertension. AB - The effect of enalapril, an angiotensin converting enzyme inhibitor, on glucose tolerance and serum insulin response to a glucose load has been evaluated in 8 non-obese patients (3 women and 5 men) with untreated essential hypertension (WHO Stage I or II) and without insulin resistance. Following a 2-month run-in control period, each patient received oral enalapril 10 mg once daily for 6 months, and an intravenous glucose tolerance test (IVGTT) was performed at the end of the run in control and active treatment periods. Treatment with enalapril significantly lowered both the systolic and diastolic blood pressures. The response of plasma glucose to the IVGTT, glucose disappearance rate (k-value) and area under the serum insulin concentration time curve were comparable between the two phases. The results suggest that long-term treatment with enalapril has no effect on glucose tolerance in non-obese, non-insulin-resistant patients with mild-to moderate essential hypertension. PMID- 8405026 TI - Effect of pinacidil on renal haemodynamics, tubular function and plasma levels of angiotensin II, aldosterone and atrial natriuretic peptide in healthy man. AB - The effects of pinacidil on renal haemodynamics, tubular function evaluated by the lithium clearance technique and the plasma levels of angiotensin II (Ang II), aldosterone (Aldo) and atrial natriuretic peptide (ANP) have been evaluated in 12 healthy volunteers given pinacidil 0.1 mg/kg IV in comparison with a placebo given to 13 different healthy volunteers. Pinacidil induced significant reductions in glomerular filtration rate (-5%), renal plasma flow (-12%), urine output (-35%), urinary sodium excretion (-20%), and the fractional excretion of sodium (-17%) and potassium (-29%). Lithium clearance and proximal and distal absolute and fractional reabsorption of sodium were not significantly changed. Ang II and Aldo were significantly increased (80% and 115%, respectively) and ANP was unchanged. The mean arterial blood pressure was not significantly changed by pinacidil, but the heart rate was increased (22%). It is concluded that bolus IV injection of pinacidil in healthy subjects reduced renal blood flow, urine volume and the urinary excretion of sodium and potassium, whereas segmental tubular function was unchanged. The increase in heart rate and activation of the renin angiotensin-aldosterone system are most likely to be secondary to stimulation of the sympathetic nervous system caused by the vasodilator effect of pinacidil. PMID- 8405027 TI - The extrapulmonary effects of inhaled hexoprenaline and salbutamol in healthy individuals. AB - We have investigated the cardiovascular and metabolic effects of multiple inhaled doses of salbutamol and hexoprenaline in 12 healthy volunteers. They inhaled 200 micrograms of salbutamol or hexoprenaline at 15 min intervals for 60 min from a metered dose inhaler (total dose 1000 micrograms). We measured heart rate, blood pressure, total electromechanical systole (as a measure of inotropic response), QTc interval on the ECG, and plasma potassium at baseline, 10 min after each inhalation, and 30 and 60 min after the last inhalation. There was no difference in the effects of the two drugs on blood pressure, total electromechanical systole, or QTc interval. Salbutamol significantly increased heart rate compared with hexoprenaline. Hexoprenaline caused a significantly greater fall in plasma potassium compared with salbutamol. PMID- 8405028 TI - Effect of concomitant administration of cimetidine and ranitidine on the pharmacokinetics and electrocardiographic effects of terfenadine. AB - Terfenadine is a widely prescribed non-sedating antihistamine which undergoes rapid and almost complete first pass biotransformation to an active carboxylic acid metabolite. It is unusual to find unmetabolised terfenadine in the plasma of patients taking the drug. Terfenadine in vitro is a potent blocker of the myocardial potassium channel. Overdose, hepatic compromise and the coadministration of ketoconazole and erythromycin result in the accumulation of terfenadine, which is thought to be responsible of QT prolongation and Torsades de Pointes ventricular arrhythmia in susceptible individuals. Cimetidine and ranitidine are two popular H2 antagonists which are often taken with terfenadine. The effects of cimetidine and ranitidine on terfenadine metabolism were studied in two cohorts of 6 normal volunteers given the recommended dose of terfenadine (60 mg every 12 h) for 1 week prior to initiation of cimetidine 600 mg every 12 h or ranitidine 150 mg every 12 h. Pharmacokinetic profiles and morning pre-dose electrocardiograms were obtained whilst the patients were on terfenadine alone and after the addition of cimetidine or rantidine. One of the subjects in each cohort had a detectable plasma level of parent compound after 1 week of terfenadine therapy alone; it did not accumulate further after addition of the H2 antagonist. The pharmacokinetics of the carboxylic acid metabolite of terfenadine (Cmax, tmax, AUC) were not significantly changed after co-administration of either H2 antagonist. None of the remaining 5 subjects in either cohort demonstrated accumulation of unmetabolised terfenadine after addition of the respective H2 antagonist and electrocardiographic QT intervals and T-U morphology in them was not changed during the course of the study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405029 TI - Factors related to QT interval prolongation during probucol treatment. AB - To clarify factors related to the QT interval prolongation produced by probucol, multivariate analysis was applied to clinical and laboratory data retrospectively obtained from 89 patients with hypercholesterolaemia, who had taken probucol for more than 3 months. The corrected QT interval (QTc) increased from 0.410s before treatment to 0.431 s after the administration of probucol; the total cholesterol level decreased from 267 mg.dl-1 to 212 mg.dl-1. None of the patients demonstrated new arrhythmia. The QTc after probucol was independently correlated with sex, serum albumin level and baseline QTc. Changes in QTc after probucol were independently correlated with the presence of ischaemic heart disease, baseline QTc, and a change in the total cholesterol level. The results suggest that a prolonged QTc is likely to appear in female patients, and in patients with a long baseline QTc or with a low serum albumin. It is also suggested that marked lengthening of the QTc is likely to occur in patients with ischaemic heart disease or with a short baseline QTc. Probucol can be used safely in patients with hypercholesterolaemia, but ECG monitoring may be necessary, especially in female patients, as well as in those with hypoalbuminaemia or with ischaemic heart disease. PMID- 8405030 TI - The effect of nitric oxide-donating vasodilators on monocyte chemotaxis and intracellular cGMP concentrations in vitro. AB - The effects of sodium nitroprusside (SNP) and 3-morpholino sydnonimine (SIN-1), isosorbide dinitrate (ISDN) and glyceryl trinitrate (GTN), and molsidomine (the inactive precursur of SIN-1) on monocyte chemotaxis and cyclic GMP (cGMP) concentration were studied. SNP and SIN-1 inhibited monocyte N-formyl-methionyl leucyl-phenylalanine-stimulated migration and increased cGMP concentrations in a dose-dependent (> 10(-5) mol.l-1) and time-dependent manner. Furthermore, 8-bromo cGMP inhibited monocyte chemotaxis in a dose-dependent fashion. In contrast, ISDN, GTN and molsidomine did not alter monocyte migration or cGMP concentration. These results support earlier observations that nitric oxide inhibits monocyte function in vitro via a cGMP-mediated mechanism. The differential effects of the spontaneous and thiol-dependent NO-donating nitrovasodilators on monocyte function suggests that monocytes, like platelets, are not able to directly metabolise ISDN and GTN. If similar observations can be made in vivo, it is possible that certain nitrovasodilators might be used therapeutically to inhibit monocyte function, for example during atherogenesis. PMID- 8405031 TI - The disposition and protein binding of batanopride and its metabolites in subjects with renal impairment. AB - We have studied the disposition of batanopride and its three major metabolites (the erythro-alcohol, threo-alcohol, and N-desethyl metabolites) in 27 subjects with various degrees of renal function after intravenous infusion of a single dose of 3.6.mg.kg-1 of batanopride over 15 min. The subjects were assigned to one of three treatment groups: group 1, normal renal function (creatinine clearance > or = 75 ml.min-1 x 1.73 m-2; n = 13); group 2, moderate renal impairment (creatine clearance 30-60 ml.min-1 x 1.73 m-2; n = 8); group 3, severe renal impairment (creatinine clearance < or = 30 ml.min-1 x 1.73 m-2; n = 6). The terminal half-life of batanopride was significantly prolonged from 2.7 h in group 1 to 9.9 h in group 3. The renal clearance of batanopride was significantly lower in group 3 (25 ml.min-1) compared with group 1 (132 ml.min-1). There were no differences in plasma protein binding or steady-state volume of distribution of batanopride among the groups. There were significantly lower renal clearances for all three metabolites in groups 2 and 3 compared with group 1. The half-lives of all three metabolites were significantly prolonged in group 3 compared with group 1. The dose of batanopride may need to be reduced in patients with creatinine clearances less than 30 ml.min-1 x 1.73 m-2 to prevent drug accumulation and avoid possible dose-related adverse effects. PMID- 8405032 TI - Similar effect of oxidation deficiency (debrisoquine polymorphism) and quinidine on the apparent volume of distribution of (+/-)-metoprolol. AB - The influence of phenotype (debrisoquine type of oxidation polymorphism) and quinidine on (+/-)-metoprolol distribution parameters was investigated in 7 young male volunteers (4 extensive and 3 poor metabolisers). (+/-)-Metoprolol tartrate 20 mg was administered as a 20 min infusion i) alone, ii) 12 h after an oral 50 mg quinidine sulphate capsule, and iii) on the last day of 3 days of treatment with 250 mg quinidine sulphate b.d. as a slow-release tablet. No stereoselectivity was apparent in either poor or extensive metabolizers. When (+/ )-metoprolol was administered alone the apparent volume of distribution at steady state (Vss) was higher in extensive than in poor metabolisers (4.84 vs 2.83 l.kg 1, respectively). Pre-treatment with low or multiple high doses of quinidine decreased Vss in extensive metabolisers to values comparable to those in poor metabolisers (3.50 and 3.18 l.kg-1, respectively), but had no significant effect in poor metabolisers (3.24 and 3.42 l.kg-1, respectively). Estimation of Vss by noncompartmental analysis or assuming elimination exclusively from the peripheral compartment yielded similar, although somewhat higher, estimates. Despite the small number of subjects, (+/-)-metoprolol distribution appeared to be different both in genetically and environmentally (quinidine)-determined poor metabolisers, and quinidine inhibition was a good, reversible in vivo model of the genetic deficiency in handling (+/-)-metoprolol. Differences both in first pass pulmonary elimination or in tissue binding are logically consistent with these observations, but the amplitude of the effect exceeds expectations from available biological evidence on selective pulmonary metabolic activity and on specific tissue binding sites. PMID- 8405033 TI - Pharmacokinetic study and effects on growth hormone secretion in healthy volunteers of the new somatostatin analogue BIM 23014. AB - We have studied the pharmacokinetics and the effects of BIM 23,014 (BIM), a new, long-acting octapeptide somatostatin analogue, on basal and stimulated GH secretion in normal men. BIM 250 micrograms sc significantly reduced a GHRH induced increase in plasma GH. The continuous sc administration of BIM for 24 h dramatically blunted spontaneous GH secretion; 2000 and 3000 micrograms daily reduced GH secretion to a greater extent than 1000 micrograms daily. During these experiments a significant negative correlation (r - 0.66) was found between plasma GH and BIM levels. Acute sc administration of 1000 micrograms BIM significantly reduced the rise in plasma GH observed in the second part of the oral glucose tolerance test. Plasma BIM levels peaked around 30 min, and the elimination half life was 90 min. Plasma BIM levels were below 1 ng/ml 6 h after the injection of 1000 micrograms BIM, and at that time GH started to rise again. We conclude that BIM 23,014 250 to 1000 micrograms sc is able to reduce the plasma GH response to GHRH or to the fall in glucose following an oral glucose tolerance test; a constant infusion of BIM, in doses 1000 micrograms daily, dramatically suppresses spontaneous GH secretion; 2000 micrograms/day by chronic subcutaneous infusion was the most effective dose of BIM in the suppression of GH secretion, and was associated only with minor adverse effects. PMID- 8405034 TI - Pharmacokinetics of oral tiopronin. AB - Ten healthy subjects were given 500 mg (3064 mumol) tiopronin, or 2 mercaptopropionylglycine (2-MPG) by mouth. Cmax was reached after 3-6 h, and after a shorter beta-phase a long terminal half-life of 53 h of total tiopronin was found. Tiopronin measured as unbound (non-protein-bound) drug disappeared more rapidly from plasma, with a calculated t1/2 of 1.8 h. Mean residence time was higher (58 h) when calculated as total tiopronin than as unbound tiopronin (6 h), and this was also the case for the volume of distribution (V lambda = 455 l vs V lambda,u = 41 l). The results indicate extensive protein binding in plasma and a deep pool of tissue bound tiopronin after the first absorption and distribution phases. Absolute bioavailability (f) was 63%, and bioavailability calculated from urinary excretion was 47%, which are well correlated with each other. Urinary excretion was mainly confined to the first 6 h (74%) and was almost complete (98%) within 12 h. We conclude that the maximal absorption of the tiopronin was late, protein and tissue binding of the drug were high and its bioavailability varied. The renal excretion of low molecular weight tiopronin occurred early, which implies that the drug should be given in divided doses, at least twice daily, for optimal efficiency in the treatment of cystinuria. PMID- 8405035 TI - Relaxant effects of mefloquine on vascular smooth muscle in vitro. AB - We have studied the effects of mefloquine on isolated rat aorta. Mefloquine (10( 8) to 1.6 x 10(-4) mol.l-1) relaxed aortic rings precontracted with both noradrenaline (10(-7) mol.l-1) and potassium chloride (60 mmol.l-1). The mefloquine-induced relaxation was somewhat attenuated by removal of the endothelium. Contractile responses to noradrenaline, potassium chloride, and calcium chloride were reduced by incubating the aortic rings in physiological salt solution containing mefloquine, suggesting that mefloquine reduces calcium influx, which is required for excitation-contraction coupling. The results show that mefloquine relaxes vascular smooth muscle via mechanisms which are partly endothelium-dependent and which is also associated with inhibition of Ca2+ influx from the extracellular medium. PMID- 8405036 TI - Pharmacokinetics of diphemanil methylsulphate in infants. AB - The pharmacokinetics of diphemanil methylsulphate was evaluated after oral administration of a single 3 mg.kg-1 dose to 5 infants being treated for symptomatic bradycardia. The mean pharmacokinetic parameters of oral diphemanil methylsulphate in infants were similar to those in adults. The mean half-life was 8.6 h. This would allow administration three times a day in infants instead of four to six times a day, as currently prescribed. The mean residence time decreases significantly with age (Spearman's r' = -1), and there is a trend for the half-life to decrease with age (r' = -0.9; NS), suggesting an influence of maturation on its elimination. PMID- 8405037 TI - Drug use in pregnancy: a comparative appraisal of data collecting methods. AB - We have compared the reliability of the information about drug therapy and pregnancy retrieved by interviewing patients with that distilled from pharmacy records. In the initial phase of each interview we used the internationally accepted open-ended technique, and extended this with an indication-oriented set of questions and then a set of specific drug-oriented questions. These data were then compared with those from pharmacy records on dispensing for the same patients during their pregnancy. The results suggest that if drug consumption during pregnancy is evaluated by interview, one should not restrict oneself to open-ended questions but should include indication-oriented and, when appropriate, drug-oriented questions. Such specific questions offer the opportunity of detecting the use of over-the-counter medication and of constructing drug use/complaint profiles. By contrast, pharmacy records will give better information in case of long recall periods and in patients with multiple and/or repeated drug use. Investigators should use the complementary elements of both techniques where appropriate. PMID- 8405038 TI - Expressions of cytokine genes during development of autoimmune sialadenitis in MRL/lpr mice. AB - Local cytokine gene expression in vivo was analyzed by direct analysis of RNA obtained from salivary gland tissues of MRL/lpr mice with autoimmune sialadenitis. The expression of cytokine genes were assessed by the reverse transcriptase polymerase chain reaction, and by immunohistochemical analysis. The expression of interleukin-1(IL-1)beta and tumor necrosis factor was detected before the onset of inflammatory lesions in the salivary glands of mice of 1 or 2 months of age, and IL-6 mRNA expression was clearly detected at the time of onset of typical autoimmune sialadenitis at 3 months of age in MRL/lpr mice, and was up regulated with advancing age. These results suggest that the overexpression of these inflammatory cytokine genes is involved in the development and progression of organ-localized autoimmunity in the salivary glands of MRL/lpr mice. PMID- 8405039 TI - Leukotriene B4 induces interleukin 5 generation from human T lymphocytes. AB - Leukotriene B4 (LTB4) has been shown to affect several interleukin (IL)-linked functions of human lymphocytes. In this study, we investigated whether LTB4 regulates IL-5 generation from human T cells and subsequently modulates eosinophil functions. Preincubation of T cells with very low concentrations (10( 12) to 10(-8) M) of LTB4 induced concentration-dependent IL-5 production, the event occurring after the first 24 h of cultivation. However, direct action of LTB4 to IL-5 generation is strictly dependent on a preincubation with appropriate concentration of LTB4. In contrast, the stereoisomer of LTB4, 5S,12S-dihydroxy 6,8,10,14-eicosatetraenoic acid showed no enhancement of IL-5 production. IL-5 released from LTB4-primed T cells elicited sustained viability of mature eosinophils and reduced the content of eosinophil cationic protein in their crystalloid matrix by degranulation. These data suggest that LTB4 induces bioactive IL-5 production from T cells and that the released IL-5 modulates eosinophil functions which might play a crucial role in eosinophil-linked allergic inflammatory process. PMID- 8405040 TI - Human CD38 is associated to distinct molecules which mediate transmembrane signaling in different lineages. AB - The CD38 antigen displays restricted functional associations with surface molecules involved in immune system and complement. Capping of the CD38 molecule in normal or neoplastic T cells is followed by rapid and specific co-modulation of the CD3-T cell receptor (TcR) complex. In normal and tumor cells of B lineage, CD38 was found to be also associated with surface Ig (sIg) and with the complement receptor 2 (CR2)/CD19 complex. The CD38 molecule expressed by purified natural killer cells displayed an association with the low affinity IgG Fc receptor (Fc gamma RIII) CD16. These observations suggest that specialized areas in the plasma membrane contain co-modulating structures, including different receptors involved in the transduction of extracellular signals. We propose a model whereby TcR, CR2 and CD16 are ligand binding structures in their respective lineages, while CD38 is a molecule involved in the intracellular transduction of the signals. PMID- 8405041 TI - Long-term thymic reconstitution by peripheral CD4 and CD8 single-positive lymphocytes. AB - Significant immigration of peripheral T cells into SCID thymus was observed following reconstitution with normal Peyer's patch, mesenteric lymph node or peripheral lymph node cells. Immunohistologic and flow cytometric analyses reveal that T cells from these tissues are found in the thymus for as long as 177 days and can account for up to 67% of intrathymic cells. The returning cells express the CD3/T cell receptor alpha/beta complex, indicative of mature cells, and are equally divided among helper (CD4+CD8-) and cytotoxic (CD4-/CD8+) phenotypes. The immigration of peripheral T cells is not accompanied by the appearance of immature, double-positive (CD4+CD8+) thymocytes as seen in similar reconstitutions using bone marrow. Taken together, these results suggest that peripheral T cells from a variety of lymphoid organs may regularly re-enter the thymus and, thus, possibly play a role in normal thymic development. PMID- 8405042 TI - Nonspecific augmentation of lymph node T cells and I-E-independent selective deletion of V beta 14+ T cells by Mtv-2 in the DDD mouse. AB - DDD/1 (DDD) mice were characterized by marked paucity of T cells in lymph nodes (LN). In DDD-Mtv-2/Mtv-2 (DDD-Mtv-2) congenics, T cells were 4- to 18-fold increased depending on ages but B cells doubled at the most. Thymus weight also increased. In DDDfDDD-Mtv-2, DDD neonatally infected with Mtv-2-derived exogenous MMTV (MMTV-2), neither LN cells nor thymus weight increased. The V beta 5+ and V beta 8+ T cell contents in LN were practically the same among three strains. The Mtv-2-induced expansion of LN T cells was polyclonal and appeared indigenous to DDD mice. Both Mtv-2 and MMTV-2 induced progressive age-dependent deletion of V beta 14+CD4+ LN cells. Mtv-2 but not MMTV-2 caused deletion of V beta 14+CD8+ LN cells and mature V beta 14+CD4+ thymocytes. Thus, Mtv-2- and MMTV-2-induced V beta 14+ T cell deletion may reflect intrathymic and peripheral elimination, respectively. The absence of I-E gene expression in DDD indicates that V beta 14+ T cell deletion advances independently of I-E molecules in this experimental system. PMID- 8405043 TI - Murine CD8+ T suppressors against mycobacterial 65-kDa antigen compete for IL-2 and show lack of major histocompatibility complex-imposed restriction specificity in antigen recognition. AB - The mechanism of antigen-specific suppression and reasons for aberrant major histocompatibility complex (MHC) class II restriction mediated by CD8+ T cells was investigated in a previously reported murine model of immunosuppression, generated by intraperitoneal priming with Mycobacterium vaccae. Both the CD4+ T helper cells (Th) and CD8+ T suppressor cell (Ts) of M. vaccae-primed mice recognized the 65-kDa antigen of the bacillus, presented by I-A and I-E, respectively. The CD8+ Ts could inhibit non-antigen-specific proliferation of primed CD4+ T cells induced by the exogenously added interleukin (IL)-2 (concanavalin A-stimulated culture supernatant). For inhibition, the Ts had to be activated by the 65-kDa antigen. The degree of inhibition was dependent upon the amount of added IL-2 and the relative numbers of primed CD8+ and CD4+ T cells. On incubation with antigen-presenting cells, and the 65-kDa antigen, the primed CD8+ T cells absorbed IL-2 as efficiently as primed CD4+ T cells. Based on this, it was concluded that the primed CD8+ T cells induced suppression by competition for IL-2. Employing the same model, the MHC restriction of recognition of the suppressor epitope of the 65-kDa antigen by the CD8+ Ts was investigated. The epitopes presented by diverse MHC class II molecules, such as self I-A, I-E and even allogeneic I-E were similar, because they were recognized by the same population of primed CD8+ Ts. Further, immunization of C57BL/6 mice with Ltk cells expressing H-2 DkKk alloantigens, stimulated CD8+ T cells capable of recognizing M.vaccae 65-kDa antigen. Based on these data, it was proposed that recognition of the suppressor epitope of the 65-kDa antigen by the primed CD8+ Ts exhibits lack of restriction specificity imposed by MHC diversity. PMID- 8405044 TI - Polymorphism of the functional immunoglobulin variable region genes in the chicken by exchange of sequence with donor pseudogenes. AB - We have isolated a number of new allelic variants of the unique functional genes encoding chicken immunoglobulin heavy and light chain variable regions (VH1 and VL1, respectively). The distribution and nature of nucleotide variation among these and previously identified VH1 and VL1 alleles demonstrates that random point mutations are likely not the predominant cause of allelic variation at these loci. Comparison of the variant nucleotides with sequences from the pseudo VH and pseudo-VL gene families, which lie 5' to VH1 and VL1, respectively, suggests that the great majority of allelic variants can be accounted for by segmental transfer of sequence from donor pseudogenes into the germ-line VH1 and VL1 genes. These results demonstrate that the chicken VH1 and VL1 genes are susceptible to sequence replacement at the germ-line level as well as somatically during antibody diversification. The limited repertoire of B cell specificities produced by gene rearrangement in the chicken has led to speculation that these specificities may play a critical role in the progression of chicken B cell development. The results presented here do not support this hypothesis since many of the allelic variant nucleotides described here encode non-conservative amino acid substitutions within the antigen-binding sites of the Ig molecule. PMID- 8405045 TI - CD45-mediated regulation of LFA1 function in human natural killer cells. Anti CD45 monoclonal antibodies inhibit the calcium mobilization induced via LFA1 molecules. AB - The TA218 and T205 monoclonal antibodies (mAb) were selected on the basis of their ability to inhibit the non-major histocompatibility complex-restricted lysis of the murine mastocytoma P815 cell line mediated by CD3-CD16+ natural killer (NK) cells. Both mAb were found to react with CD45 molecules, as demonstrated by immunoprecipitation after surface iodination and western blot analysis. A panel of tumor target cells susceptible to lysis by polyclonal or clonal CD3-CD16+ NK cells was used to study the mAb-mediated inhibitory effect. The inhibition of cytolysis mediated by TA218 and T205 mAb was found to consistently parallel the inhibition mediated (with the same tumor target cells) by the anti-LFA1 alpha mAb TS.1.22 or by the anti-LFA1 beta mAb TS.1.18. However, different from the anti-LFA1 mAb, T205 or TA218 mAb did not inhibit the binding of activated CD3-CD16+ effector NK cells to the same tumor target cells. This finding supported the concept that the anti-CD45 mAb-mediated inhibition could occur at a post-binding stage. In polyclonal or clonal CD3-CD16+ NK cells T205 or TA218 mAb were found to reduce by 50-70% the intracellular Ca++ ([Ca++]i) mobilization induced by anti-LFA1 alpha or anti-LFA1 beta mAb. On the other hand, TA218 and T205 mAb did not inhibit the Ca++ mobilization induced by anti-CD16 mAb or phytohemagglutinin, thus suggesting that, in NK cells, CD45 molecules may exert a selective inhibitory effect on the signal transduction mediated by LFA1 molecules. In line with this hypothesis, the cytolytic activity of human NK clones was triggered in the presence of the hybridoma cells secreting either anti CD16 or anti-LFA1 alpha mAb (as "triggering targets"). This effect of anti-LFA1 alpha, but not of anti-CD16 hybridoma was susceptible to inhibition by the anti CD45 mAb T205 or TA218. Further, experiments on cloned NK cells indicated that T205 or TA218 mAb induced a strong decrease in the constitutive phosphorylation of the LFA1 alpha chain (but not of HLA class I antigens). Taken together, these studies suggest that in human NK lymphocytes, CD45 molecule may regulate both the activation state and the function of the LFA1 molecule. PMID- 8405046 TI - Murine platelet endothelial cell adhesion molecule (PECAM-1)/CD31 modulates beta 2 integrins on lymphokine-activated killer cells. AB - Lymphokine-activated killer (LAK) cells are able to colonize sites of tumor lesions in mouse and man. The molecular mechanisms of homing in on tumors are largely unknown. However, before LAK cells can reach the tumor, they must adhere to the vascular endothelial within the lesion and then extravasate. We developed a novel mAb, EA-3, which recognizes the murine homologue of the human adhesion molecule CD31. It is present on a subpopulation of murine LAK cells and all endothelial cells. CD31 was also involved in the adhesion of LAK cells to endothelium. Since CD31 can initiate integrin activation by inside-out signaling after binding to its ligand, EA-3 was used to minimic this in adhesion assays. It induces modifications in the beta 2 integrin LFA-1, leading to increased binding capacities of the cells to endothelium. In contrast, beta 1 integrins and RGD binding integrins were not affected. These results suggest that expression of CD31 might confer adhesive advantages for LAK cells prone to tumor infiltration. PMID- 8405047 TI - Kinetic study of interleukin-2 binding on the reconstituted interleukin-2 receptor complexes including the human gamma chain. AB - To gain an insight into roles of the interleukin-2 (IL-2) receptor gamma chain in IL-2 binding mechanisms, we examined association and dissociation rate constants for IL-2 binding with a series of fibroblastoid L929 cell lines expressing IL-2 receptor complexes reconstituted by transfection with the alpha, beta and gamma genes. The association rate constant (k) with the alpha beta gamma complex on L cells was fourfold larger than that with the alpha beta complex on L cells, and the dissociation rate constant (k') was one fifth of that with the alpha beta complex. These results indicate that the gamma chain is involved in both mechanisms by which IL-2 associates with and dissociates from receptors, resulting in the generation of the high-affinity IL-2 receptor along with the alpha and beta chains. During the course of this study, we found that the IL-2 dissociation from alpha beta gamma complex on lymphoid cells was a lot slower than that from the alpha beta gamma complex on fibroblast cells. A similar difference was observed with the beta gamma complex. These observations may indicate functional and constitutional differences of the high- and intermediate affinity IL-2 receptor complexes between lymphoid and fibroblastoid cells. PMID- 8405048 TI - Interferon-gamma regulation of C4 gene expression in cultured human glomerular epithelial cells. AB - The fourth component of human complement is mainly produced in liver, but extrahepatic gene expression has been reported in human renal tissue and other tissues, and is thought to contribute to the inflammatory reaction in local tissue. To identify the cellular origin of C4 synthesis in human kidney, we studied C4 gene expression and regulation by interferon-gamma (IFN-gamma) and C4 protein biosynthesis in isolated glomerular epithelial cells. cDNA/polymerase chain reaction analysis showed that C4 transcripts are present in glomerular epithelial cells (GEC), and that gene expression is up-regulated by IFN-gamma. Metabolic labeling studies showed that GEC synthesize and secrete C4 as three polypeptide chains of approximately 90, 70, and 30 kDa, which correspond to the bands produced by hepatoma cells. These results suggest that the fourth component of complement is produced by GEC. The GEC has an important role in the maintenance of glomerular barrier function, which is lost in a number of complement-dependent conditions; glomerular epithelial synthesis of C4 could have a bearing on the possible physiological or pathological roles of complement under different circumstances. PMID- 8405049 TI - Evidence for programmed cell death of self-reactive gamma delta T cell receptor positive thymocytes. AB - The negative selection of T cells expressing the gamma delta T cell antigen receptor (gamma delta T cells) was studied using transgenic mice expressing a gamma delta receptor with specificity for an H-2T-linked class I major histocompatibility complex molecule from H-2b mice. The potentially self-reactive gamma delta thymocytes in H-2b/d transgenic mice are larger and have lower levels of gamma delta T cell receptor expression than gamma delta thymocytes from H-2d mice. H-2b/d gamma delta thymocytes do not respond to H-2b antigen-presenting cells, and thus are inactive compared to H-2d gamma delta thymocytes. However, the H-2b/d gamma delta thymocyte population, but not the H-2d gamma delta thymocyte population, undergoes a high rate of programmed cell death when placed in overnight culture. These observations constitute the first direct evidence that self-reactive gamma delta thymocytes undergo programmed cell death. This in vitro programmed cell death of self-reactive gamma delta thymocytes may reflect the clonal deletion process that results in a depletion of gamma delta T cells in the peripheral lymphoid organs of adult H-2b/d mice. We also present evidence that self-reactive gamma delta T cells, similarly to alpha beta T cells, undergo a lesser degree of clonal deletion in neonatal mice compared to adult mice. PMID- 8405050 TI - Association of the p56lck protein tyrosine kinase with the Fc gamma RIIIA/CD16 complex in human natural killer cells. AB - The multimeric Fc gamma RIIIA (CD16) complex is expressed on the surface of natural killer (NK) cells and is composed of a 50-70-kDa transmembrane glycoprotein Fc gamma receptor (CD16), the T cell receptor (TCR)-zeta chain, and the Fc epsilon RI gamma chain. Cross-linking Fc gamma RIIIA initiates the rapid tyrosine phosphorylation of multiple substrates including the zeta subunit and causes subsequent cell activation and antibody-dependent cellular cytotoxicity (ADCC). The subunits of the Fc gamma RIIIA complex lack intrinsic protein tyrosine kinase (PTK) activity, suggesting that receptor-induced tyrosine phosphorylation events are mediated by a nonreceptor PTK. We report here that the human Fc gamma RIIIA is complexed with p56lck, a src-family PTK previously found associated with the CD4 and CD8 receptors on T cells. Upon engagement of the CD16 receptor, p56lck is rapidly (within 30 s) and transiently phosphorylated on tyrosine residues. Several Fc gamma RIIIA-associated proteins are identified in immune complex kinase assays including the TCR-zeta subunit, a p70-90 zeta associated protein (ZAP), p50a (acidic) and p50b (basic), and p56lck. We demonstrate that the src-family protein tyrosine kinase inhibitor, herbimycin A, blocks increased intracellular calcium levels and ADCC caused by CD16 cross linking on NK3.3 cells. Likewise cross-linking CD16 with the protein tyrosine phosphatase CD45, abrogates CD16-induced calcium mobilization. These data suggest that p56lck is physically associated with Fc gamma RIIIA (CD16) and functions to mediate signaling events related to the control of NK cellular cytotoxicity. PMID- 8405051 TI - Signaling via CD28 costimulates lymphokine production, but does not reverse unresponsiveness to interleukin-2 in anti-CD3 triggered Th1 cells. AB - Previously, it has been described that the ability of murine Th1 cells to proliferate in response to exogenous interleukin (IL)-2 is blocked when these cells are exposed to immobilized anti-CD3 antibodies. In the present study we examined whether simultaneous triggering of the T cell antigen CD28 can prevent the induction of unresponsiveness to IL-2 in Th1 cells. We report that costimulation of Th1 cells with anti-CD28 monoclonal antibodies (mAb) did not overcome unresponsiveness to IL-2 induced by various amounts of immobilized anti CD3 antibodies. However, stimulation with anti-CD28 mAb strongly augmented IL-2 and interferon-gamma production in anti-CD3-exposed Th1 cells. Thus, despite the fact that anti-CD28 mAb is a potent costimulus for lymphokine production, signaling through CD28 does not seem to be sufficient to trigger proliferation in Th1 cells activated via the T cell receptor. These data suggest the existence of at least three signals to trigger Th1 cell activation. The first is mediated by ligation of the T cell receptor. One cosignal, delivered by the CD28 molecule, leads to IL-2 production. A third, still undefined, signal is required for proliferation in response to IL-2. PMID- 8405052 TI - Surface phenotype and functions of tumor-infiltrating dendritic cells: CD8 expression by a cell subpopulation. AB - Although the function and significance of tumor-infiltrating dendritic cells (TIDC) in the immune response to tumor have never been clearly demonstrated, their location suggests that they play a critical role in the presentation of tumor antigen to specific T cells. We studied the morphological and functional characteristics of interstitial dendritic cells (DC) located inside tumors obtained by injection of cancer cells into syngeneic rats. Single and double immunostaining of tumor sections revealed a dense network of cells which expressed class II major histocompatibility complex (MHC II) molecules. Cell morphology and surface markers were characteristic of DC populations in other tissues. These DC were in close contact with tumor cells and increased in number as the tumor grew larger. Unexpectedly, a subpopulation of morphologically characteristic TIDC expressed both CD8 and MHC II molecules. TIDC were purified from tumors by gradient centrifugation and immunobeads and characterized by morphology, ultrastructural study and surface markers studied by flow cytometry. TIDC were negative for the CD5 molecule (a pan T cell marker), and were not labeled with 3.2.3 monoclonal antibody (mAb) (an NK cell marker) or with Ki-M2R mAb (a macrophage marker). A subpopulation of TIDC expressed the CD8 molecule, confirming the in situ results. TIDC expressed high levels of class I and class II MHC molecules and the adhesion molecule ICAM-1. This expression is compatible with effective antigen presenting function. Purified TIDC triggered rapid and high levels of proliferation of tumor-immune T cells in vitro, demonstrating the potential of these cells to constitutively process and present tumor-associated antigens. PMID- 8405053 TI - In vivo administration of major histocompatibility complex class I-specific peptides from influenza virus induces specific cytotoxic T cell hyporesponsiveness. AB - We have been investigating the immunogenicity of two class I major histocompatibility complex-specific peptides with a sequence derived from influenza virus nucleoprotein specific for Kd and one for Db. Peptide-modified splenocytes are unable to immunize for a primary cytotoxic T (Tc) cell response in vivo, or secondary response in vitro. Peptide-modified stimulator cells can boost virus-primed splenocytes for a strong secondary response in vitro. Animals primed with syngeneic peptide-modified splenocytes upon challenge with virus in vivo do not generate strong secondary Tc cell responses on day 3 after challenge in contrast to virus primed animals. Day 6 responses of virus-challenged, peptide primed animals are reduced as compared to unprimed mice. This hyporesponsiveness is independent of CD8+ T cells in the priming population and can be elicited with tumor cell lines. The data are discussed in the framework of the two-signal model of immune induction. PMID- 8405054 TI - The G protein-coupled receptor BLR1 is involved in murine B cell differentiation and is also expressed in neuronal tissues. AB - The BLR1 gene, isolated initially from Burkitt's lymphoma cells (Eur. J. Immunol. 1992. 22: 2795), encodes a G protein-coupled receptor with significant relationship to receptors for chemokines (IL-8, MIP-1 alpha) and neuropeptides. The murine homologue of human BLR1 was cloned and used to investigate its expression in vivo. blr1-specific transcripts are observed in secondary lymphatic organs and to a lesser extent in brain of adult mice but not in other tissues. RNA in situ hybridization localizes blr1 transcription to primary follicles and to the mantle zone of secondary follicles. SCID mice in which mature B cell development is severely impaired exhibit a strongly reduced level of blr1 specific RNA in the spleen. The analysis of murine lymphoid tumor cell lines representing distinct stages of the B cell lineage reveals elevated expression of blr1 in B cell lymphomas but not in pre-B lymphomas or plasmacytomas. Induction of differentiation of resting B cells by cytokines or mitogens down-regulates expression of blr1. RNA in situ hybridization using brain sections of adult mice detects blr1 transcription in the granule and Purkinje cell layer of the cerebellum. Interestingly, the blr1 gene is also expressed during late embryogenesis in fetal liver and brain. In view of the remarkable expression pattern in the B cell lineage we suggest that murine BLR1 may represent a cytokine/neuropeptide receptor exerting regulatory functions on recirculating mature B lymphocytes. PMID- 8405055 TI - Migration patterns of thymus-derived gamma delta T cells during chicken development. AB - Cell transfer experiments in congenic chick strains, one of which expresses the ov antigen marker, indicate that intestinal gamma delta T cells are derived from gamma delta+ thymocytes in embryos and newly hatched birds, and this early intestinal colonization occurs in two discrete waves. Here, we extend these studies to show that splenic colonization by gamma delta T cells occurs in essentially the same way. Following the engraftment of ov+ thymic lobes in thymectomized ov- recipients, gamma delta T cells migrate both to the spleen and intestine. By 1 week after hatching, a third generation of thymus-derived gamma delta T cells begins to migrate to both peripheral lymphoid organs, and this thymus-dependent seeding process is sustained over the first weeks of life. The survival time for splenic gamma delta migrants is significantly less than for the intestinal migrants. Tissue section analysis indicates that gamma delta T cells enter the intestinal epithelium at all villus levels. A shift in the gamma delta intraepithelial lymphocyte distribution toward the villus tip in thymectomized birds suggests the comigration of enterocytes and gamma delta intraepithelial lymphocytes. However, survival kinetics of the donor gamma delta population and a relatively high division rate of intestinal gamma delta T cells indicate that founder thymic migrants produce relatively long-lived clones of intestinal gamma delta T cells. PMID- 8405056 TI - Dynamic response of murine gut intraepithelial T cells after infection by the coccidian parasite Eimeria. AB - The response of murine intraepithelial lymphocyte (IEL) populations to challenge by Eimeria vermiformis, a naturally occurring protozoan parasite of the gut epithelium, has been studied. The number of recoverable IEL increased within 3 days post infection, was depleted by day 7 post infection, but was significantly increased again by about day 14 post infection. Special attention was paid to gamma delta+ IEL T cells, because they are of unknown functions. These cells showed changes in numbers similar to the total IEL population. Moreover, by day 3, increased expression was detected among gamma delta+ IEL T cells, of T cell receptor genes not constitutively associated with the intestine. These results demonstrate that the IEL repertoire, and within that, the gamma delta+ T cell repertoire, can be extremely dynamic post infection with a naturally occurring epithelial-tropic pathogen. In considering the potential benefits of such IEL changes, we hypothesize that they may be relevant to the transient protection of the host's epithelium, both from parasitic re-infection, and from potentially damaging inflammation. PMID- 8405057 TI - Ligation of CD28 receptor by B7 induces formation of D-3 phosphoinositides in T lymphocytes independently of T cell receptor/CD3 activation. AB - The co-stimulatory role of B7/CD28 interactions is important in promoting T cell activation. Very little is known about the intracellular events that follow CD28 engagement although recent evidence has implicated coupling of CD28 to a protein tyrosine kinase signal transduction pathway. In this study we have investigated the putative role of D-3 phosphoinositides as mediators of CD28 receptor signaling, since phosphoinositide (PI) 3-kinase, the enzyme responsible for D-3 phosphoinositide formation, is a known substrate for protein tyrosine kinases associated with certain T cell surface receptors such as CD4 and interleukin-2 receptor. The lipid products of PI 3-kinase activity have been suggested to play a role in mitogenic signaling and growth regulation in other cells. Chinese hamster ovary cells (CHO) previously transfected with B7 cDNA, induced time dependent elevation above basal levels of phosphatidylinositol(3,4)-bisphosphate (PtdIns(3,4)P2) and PtdIns(3,4,5)P3, while parental CHO cells that did not express B7 had no effect on these lipids. Moreover, the elevation of these same lipids by CD3 ligation was potentiated in an additive manner by CHO-B7+ but not by CHO-B7- cells. CHO-B7+ and CHO-B7- cells did not activate phospholipase C as evidenced by their inability to modulate basal or CD3-induced changes in the levels of phosphatidic acid or D-4 and D-5 phosphoinositides. These data imply that PI 3-kinase but not phospholipase C, may be an important signal transduction molecule with respect to CD28-mediated co-stimulation and T cell activation following ligation by B7. PMID- 8405058 TI - T cell development in a major histocompatibility complex class II-deficient patient. AB - In this report we show that the major histocompatibility complex (MHC) class II negative thymus of a bare lymphocyte syndrome (BLS) patient contains a reduced CD4+ CD8- T cell population when compared to thymocytes derived from a MHC class II-expressing thymus. Of these CD4+ CD8- BLS thymocytes, approximately only one third co-expressed the CD3 antigen, moreover at a lower expression level when compared to control thymocytes. This suggests a partial maturation of the CD4+ CD8- T cells in the absence of MHC class II expression. Among the BLS thymocytes, CD4+ CD8+ thymocytes could easily be detected. Noteworthy, the number of CD4- CD8+ thymocytes was significantly increased. CD4+ CD8- T cells could also be found among the BLS peripheral blood mononuclear cells, albeit at reduced numbers. Despite the absence of peripheral MHC class II expression, the majority of these CD4+ CD8- T cells co-expressed the CD45RO marker. In the BLS patient, thymocytes as well as peripheral CD4+ CD8- T cells were not restricted in the use of the available T cell receptor (TcR) V gene family pool. However, the lack of detectable levels of thymic and peripheral MHC class II antigen expression in the BLS patient had altered the CD4-skewing patterns of TcR V gene families which were present in normal individuals. In conclusion, the lack of MHC class II expression in the BLS patient does not completely inhibit the CD4+ CD8- T cell development. PMID- 8405059 TI - Viral hemagglutinin augments peptide-specific cytotoxic T cell responses. AB - In attempt to increase the induction of peptide-specific cytolytic T cells (CTL) we investigated the effect of the Newcastle disease virus (NDV) hemagglutinin neuraminidase (HN) gene product on the activation of peptide-specific CTL. Spleen cells of CH3 mice immunized against the influenza nucleoprotein peptide 50-63 (NP 50-63) were restimulated in vitro (i) with peptide-pulsed syngeneic fibroblast cells (Ltk-) as antigen-presenting cells, which were in addition (ii) infected with NDV or (iii) stably transfected with the HN cDNA of NDV. A greater than sixfold increase in peptide-specific CTL responses was observed in cultures restimulated with peptide-pulsed Ltk- cells which co-expressed viral hemagglutinin due to either infection or transfection. A similar augmentation was seen in CTL responses against other types of antigen (major histocompatibility complex alloantigens, minor histocompatibility antigens or tumor antigens) when suboptimal cultures were stimulated with the respective antigen-presenting cells modified by NDV infection. These findings suggest that NDV or viral HN expressed on antigen-presenting cells or tumor cells can exert a T cell co-stimulatory function. PMID- 8405060 TI - An age-related gamma delta T cell suppressor activity correlates with the outcome of autoimmunity in experimental Trypanosoma cruzi infection. AB - In this work the suppressive activity of splenic T cells from young and aged BALB/c mice infected with Trypanosoma cruzi were compared and correlated with the development of autoimmune myocarditis. The T cells from young adult BALB/c mice with acute T. cruzi infection exhibit suppressor activity when added to full allogeneic or Mls-disparate mixed lymphocyte cultures. This suppression could not be reverted by exogenous interleukin (IL)-2 and was not directly dependent on the presence of IL-4, IL-10 or transforming growth factor-beta. Further characterization of the T cell lineage responsible for the suppressor activity by in vitro and/or in vivo depletion with monoclonal antibody to alpha beta or gamma delta T cell receptor revealed that splenic gamma delta T cells function as suppressor lymphocytes in young T. cruzi-infected mice. In addition, these young adult BALB/c mice do not develop autoimmune myocarditis and showed a low incidence of syngeneic heart graft rejection in the early chronic phase of the infection. In contrast, T cells from acutely infected aged BALB/c mice lacked demonstrable T suppressor activity. Furthermore, these mice developed a severe autoimmune myocarditis as early as 2 months after the onset of the infection, when the majority of them reject syngeneic heart grafts. These findings suggest that a gamma delta T cell-mediated suppressor mechanism may operate in the avoidance of the breaking of tissue-specific tolerance during the acute infection. Moreover, such a mechanism is likely related to the immune system chronobiology. PMID- 8405061 TI - Immunization against tumor and minor histocompatibility antigens by eluted cellular peptides loaded on antigen processing defective cells. AB - Material eluted from RMA lymphoma or B6 spleen cells under acid conditions was fractionated by reverse phase high-performance liquid chromatography, and tested for ability to restore the sensitivity to cytotoxic T lymphocytes of the processing/presentation defective mutant line RMA-S. This allowed identification of three fractions (termed M1, M2 and M3) carrying B6 antigens recognized by cytotoxic T lymphocytes (CTL) elicited across the minor histocompatibility barrier A.BY anti-B6 (both H-2b) and one fraction (termed T1) carrying a tumor antigen recognized by B6 anti-RMA CTL. By parallel runs of material from cell lysates over major histocompatibility complex class I affinity columns, the M2 and M3 antigens were defined as Kb restricted, and M1 and T1 as Db restricted. Isolated fractions loaded onto RMA-S cells could be used to prime anti-minor histocompatibility antigen and tumor CTL in vivo. They could also be used for in vitro restimulation of spleen cells from mice that had been primed either by antigen-loaded RMA-S, or by wild-type RMA tumor cells and B6 splenocytes. The CTL generated by these methods were specific for the loading antigen, and they also recognized the antigen on the "physiological" target, i.e. RMA or B6 lymphoblasts. This system based on RMA-S as an immunization and target antigen reporter cell may be used for dissection of complex CTL responses, e.g. in studies of clonal composition and epitope dominance, or for studies of tumors that are poor stimulators of immunity. PMID- 8405062 TI - In vitro negative selection of alpha beta T cell receptor transgenic thymocytes by conditionally immortalized thymic cortical epithelial cell lines and dendritic cells. AB - We have established conditionally immortalized thymic cortical epithelial cell lines from transgenic mice carrying a temperature-sensitive SV40 large T antigen. One of these cell lines expresses cortical markers and produces IL-1 alpha, IL-6, IL-7, and TGF-beta 1. These cells express class I major histocompatibility complex (MHC) constitutively and class II MHC upon induction with IFN-gamma. The cells appear to have a normal class I antigen presenting pathway since messages for both peptide transporter genes (TAP1, TAP2) were detected. The ability of these cortical epithelial cells to present peptide antigen was compared to that of thymic dendritic cells. In suspension culture with alpha beta T cell receptor (TcR) transgenic thymocytes, these epithelial cells and dendritic cells (pre pulsed with peptide cognate for the transgenic TcR) caused down-regulation of CD4, CD8, and TcR in an antigen dose-dependent and MHC-restricted manner. CD4dullCD8dull cells were taken as evidence for negative selection because these cells contained apoptotic DNA. Concentration of peptide required for negative selection of thymocytes was similar between dendritic cells and cortical epithelial cells. In contrast, alpha beta TcR transgenic spleen cells were activated only by dendritic cells but not by cortical epithelial cells. PMID- 8405063 TI - Early B cell development requires mu signaling. AB - In vitro studies with Abelson murine leukemia virus (AMuLV)-transformed murine pre-B cell lines demonstrated that wild-type mu but not mutant mu chains lacking the first constant domain (mu delta 1) can efficiently induce Ig light (L) chain gene rearrangement. Using antibodies against the cytoplasmic tail of the immunoglobulin co-receptor beta (Ig beta) chain we find mu, but not mu delta 1 chains associated with Ig beta. Since a heterodimer of surface-labeled proteins was co-precipitated with mu we conclude that only wild-type mu is associated with the Ig alpha/Ig beta co-receptor on the surface of pre-B cell lines. Mutant mu delta 1 chains achieve their surface expression by utilizing a glycophospholipid anchor. In vivo analysis of transgenic mice expressing either mu or mu delta 1 transgenes revealed the expected "normal" B cell development in the case of wild type mu transgenic lymphocytes, but a block in differentiation of mu delta 1 transgenic lymphocytes. The maturation block occurs at the developmental transition of pre-B lymphocytes from the CD43/S7+, CD45R/B220low stage to the CD43/S7-, B220low/high stage in which the majority of L chain gene rearrangements occur. These results, together with the observed inability of the mu delta 1 chains to signal activation of L chain gene joining and to associate Ig alpha/Ig beta in pre-B cell lines suggests that signals mediated by the protein complex composed to mu/Ig alpha/Ig beta are crucial during differentiation of pre-B lymphocytes. PMID- 8405064 TI - Molecular cloning of a ligand for the inducible T cell gene 4-1BB: a member of an emerging family of cytokines with homology to tumor necrosis factor. AB - 4-1BB is an inducible T cell antigen that shows sequence homology to members of an emerging family of cytokine receptors, including those for tumor necrosis factor and nerve growth factor. To aid in the analysis of the function of 4-1BB we have utilized a soluble form of the molecule as a probe to identify and clone the gene which encodes its ligand. The ligand for 4-1BB is a type II membrane glycoprotein that has homology to tumor necrosis factor, lymphotoxin, and the ligands for CD40 and CD27, all of which are themselves ligands to receptors in this superfamily. The gene for 4-1BB is on mouse chromosome 4 and maps close to the p80 form of the tumor necrosis factor receptor as well as the gene for CD30. The gene for 4-1BB ligand maps to mouse chromosome 17, but considerably distal to the tumor necrosis factor and lymphotoxin genes. Interaction of 4-1BB with its ligand induces the proliferation of activated thymocytes and splenic T cells, a response which is mimicked on similar cell populations stimulated with an antibody to 4-1BB. PMID- 8405065 TI - The effect of bone marrow and thymus chimerism between non-obese diabetic (NOD) and NOD-E transgenic mice, on the expression and prevention of diabetes. AB - The non-obese diabetic (NOD) mouse is an established animal model of the autoimmune disease, insulin-dependent diabetes mellitus (IDDM). The NOD-E mouse is a transgenic mouse which expresses the I-E molecule (absent in NOD mice). Expression of I-E protects these mice from both insulitis and IDDM. We have investigated the possible mechanisms of this protection by constructing bone marrow, and combined bone marrow and thymus chimeras between NOD and NOD-E mice. Our data suggest that thymic epithelium may play no direct role in either protection against, or promotion of, IDDM. Protection from diabetes is provided either by NOD-E donor bone marrow or NOD-E recipient non-thymic radioresistant cells. The means by which protection may be achieved in this system are discussed. PMID- 8405066 TI - Complementarity-determining region 2 is implicated in the binding of staphylococcal protein A to human immunoglobulin VHIII variable regions. AB - Staphylococcal protein A (SPA) has two distinct binding sites on human immunoglobulins. In addition to binding to the Fc region of most IgG molecules, an "alternative" binding site has been localized to the Fab region of human immunoglobulins encoded by heavy chain variable gene segments belonging to the VHIII family. Comparison of amino acid sequences of closely related SPA-binding and -non-binding proteins suggested that VHIII-specific residues in the second complementarity-determining region (CDR2) were likely responsible for SPA binding activity. Site-directed mutagenesis of a single amino acid residue in CDR2 converted an IgM rheumatoid factor which did not bind SPA to an SPA binder. These findings, therefore, locate a critical site involved in SPA binding to the CDR2 of human immunoglobulins encoded by VHIII family gene segments. PMID- 8405067 TI - Divergent effects of interleukin-10 on cytokine production by mononuclear phagocytes and endothelial cells. AB - Interleukin-10 (IL-10), a product of T helper type 2 (TH2) cells and monocytes, inhibits cytokine production in mononuclear phagocytes. Given the similarities and interrelationship between cells of the monocyte-macrophage lineage and endothelial cells, we examined the effects of IL-10 on vascular endothelium. Murine IL-10 induced low levels of IL-6 production and amplified induction of IL 6 by lipopolysaccharide (LPS) or IL-1 in the murine tEND.1 endothelioma line, used for these studies because it retains properties of normal endothelium. The effect was more evident after prolonged (48-72 h) exposure to IL-10. IL-10 had similar activity on other endothelioma lines, whereas it inhibited IL-6 production by peritoneal macrophages. Induction and amplification of cytokine production by IL-10 was associated with higher levels of mRNA, which were maintained longer (up to 48 h) than in controls. In addition to IL-6, murine IL 10 induced or amplified expression of the chemoattractant cytokines monocyte chemotactic protein-1 (MCP-1) and KC. Human IL-10 inhibited IL-6 release by LPS stimulated human peripheral blood mononuclear cells, whereas it did not interfere with cytokine production by LPS- or IL-1-stimulated human umbilical vein endothelial cells. The selective inhibitory action of IL-10 on mononuclear phagocytes versus endothelial cells may play a role in the pathophysiology of TH2 directed responses. PMID- 8405068 TI - Staphylococcal enterotoxin B up-regulates interleukin-2 receptor beta chain expression on tonsillar B cells. AB - The superantigen staphylococcal enterotoxin B (SEB) selectively up-regulates the interleukin-2 receptor (IL-2R) beta chain (p70) without up-regulating the IL-2R alpha chain (CD25) on a human tonsillar B cell population depleted of T cells. This action of SEB, probably mediated by binding to major histocompatibility complex class II, renders B cells sensitive to T cell-derived IL-2 and is sufficient for induction of vigorous DNA synthesis with low concentrations of IL 2. This explains one of the mechanisms by which bacterial superantigens activate large numbers of B cells and may reflect a similar mechanism operative in cognate helper T cell/B cell interactions. PMID- 8405069 TI - Interleukin-10 inhibits the induction of monocyte procoagulant activity by bacterial lipopolysaccharide. AB - Monocytes stimulated with bacterial lipopolysaccharide (LPS) generate a procoagulant activity (PCA) related to the induction of tissue factor (TF) expression at their surface. Since interleukin-10 (IL-10) was recently shown to inhibit LPS-induced cytokine production and is currently considered as a potential therapeutic agent in septic shock, we were interested to determine its effects on LPS-induced monocyte PCA. Peripheral blood mononuclear cells (PBMC) from healthy donors were incubated with 1 microgram/ml LPS in the presence of serial dilutions of recombinant human IL-10 and PCA was determined after 6 h in a one-stage clotting assay. IL-10 inhibited in a dose-dependent manner LPS-induced TF-dependent PCA: a significant effect was already observed with 30 pg/ml IL-10 while 64-97% inhibition was achieved with 120 pg/ml IL-10. In parallel flow cytometry experiments, IL-10 was shown to block LPS-induced TF expression at the surface of monocytes. In order to inhibit LPS-induced PCA, IL-10 had to be added to PBMC at least 6 h before LPS challenge. This inhibitory effect of IL-10 was already apparent at the TF mRNA level and was prevented by co-incubation with cycloheximide (20 micrograms/ml). These data suggest that IL-10 acts via the induction of protein(s) which might interfere with TF gene transcription or mRNA stability. We conclude that the protective effects of IL-10 in endotoxinemia might be related not only to cytokine synthesis blockade but also to inhibition of LPS-induced PCA. PMID- 8405070 TI - Regional sublocalization of the human CD69 gene to chromosome bands 12p12.3 p13.2, the predicted region of the human natural killer cell gene complex. AB - The early activation antigen CD69 is a member of a supergene family of type II integral membrane proteins with a C-type lectin domain. In recent reports the genes encoding the natural killer (NK) cell-related molecules of this supergene family, NKR-P1, NK1.1 and Ly-49, were shown to be clustered in a chromosomal region in mouse, termed the NK gene complex. The human homologue of this complex is likely to reside on chromosome 12 near the PRP locus (12p13.2). By analyzing T cell hybrids, the CD69 gene was previously mapped to human chromosome 12. Here we report the regional sublocalization of the human CD69 gene to chromosome bands 12p12.3-p13.2, suggesting that CD69 belongs to one linkage group together with different cell surface molecules on NK cells. PMID- 8405071 TI - Mouse intestinal epithelial cells express the self superantigen Mls1a. AB - In previous studies, we demonstrated that intestinal epithelial cells of the mouse small intestine could present exogenous antigen to specific CD4+ T cell hybridomas. We now report on the ability of normal enterocytes to present the self superantigen Mls1a. Enterocytes from Mls1a but not from Mls1b strains stimulated interleukin-2 production through a V beta 6+ T cell hybridoma specific for Mls1a determinants. Antibody inhibition experiments showed that enterocytes presented Mls determinants via a major histocompatibility complex class II dependent mechanism. Furthermore, the ability of enterocytes to activate V beta 6+ Mls1a-specific T cells was inhibited by monoclonal antibodies against the Orf protein encoded by an Mtv-7 provirus which is associated with Mls1a expression. These findings provide evidence for the first time that Mls determinants are expressed on normal enterocytes and support the theory of a possible role of these cells in extrathymic selection of T cell receptor V beta repertoire of intraepithelial T lymphocytes. PMID- 8405072 TI - Nicotinic receptor agonists exhibit anxiolytic-like effects on the elevated plus maze test. AB - The effects of nicotinic receptor agonists on the elevated plus-maze test of anxiety were investigated in CD1 mice after intraperitoneal injections. Nicotine and lobeline, but not cytisine, exhibited a significant increase in the time spent by the mice in the open arms, a measure of anxiolytic activity. Nicotine also increased the total number of arm entries, a measure of general activity, but this effect was secondary to its anxiolytic-like properties. Nicotinic receptor antagonists on their own did not modify the behavior of mice in the maze. The effect of nicotine was mediated by central nicotinic receptors as it was blocked by the centrally-acting nicotinic antagonists mecamylamine and chlorisondamine, but not by hexamethonium (a peripherally acting blocker). Cotinine, the major metabolite of nicotine, was evaluated at different times after systemic injections and had no effect in the plus-maze. The anxiolytic-like profile induced by nicotinic receptor stimulation was not associated with potentiation of alcohol effects, a liability associated with the benzodiazepine therapy. This study demonstrates the anxiolytic-like properties of nicotine and lobeline in mice, and suggests that central nicotinic receptors are involved in the expression of emotional behavior. PMID- 8405073 TI - Protective effects of calcitonin gene-related peptide in different experimental models of gastric ulcers. AB - Intravenous administration of calcitonin gene-related peptide (CGRP) prevented in a dose-dependent manner reserpine-induced gastric mucosal damage, but failed to affect the lesions produced by ethanol administration. In pylorus-ligated rats, CGRP significantly reduced gastric volume, total acid and peptic activity output as well as ulcer formation. These protective effects of CGRP were not present when rats were pretreated with cysteamine. Our data suggest that CGRP exerts its antisecretory and antiulcer activity, at least in part, by interfering with somatostatin transmission. PMID- 8405074 TI - Impairment of working memory induced by scopolamine in rats with noradrenergic DSP-4 lesions. AB - In a working memory task with a three-panel runway set-up, DSP-4 (N-(2 chloroethyl)-N-ethyl-2-bromobenzylamine), a noradrenergic neurotoxin, at 50 mg/kg i.p. given 14 days before testing, had no effect on the number of errors (attempts to pass through two incorrect panels of the three-panel gates at four choice points). Working memory errors were significantly increased by scopolamine, a muscarinic receptor antagonist, at 0.32 mg/kg i.p. given 20 min before testing, whereas errors were not affected by the 0.1 mg/kg dose. In rats with noradrenergic DSP-4 lesions, 0.1 mg/kg scopolamine significantly increased the number of working memory errors. However, DSP-4 at 50 mg/kg and scopolamine at 0.1 mg/kg whether they were administered alone or in combination had no effect on reference memory errors. These results suggest that noradrenergic deficits aggravate the working memory impairment induced by blockade of muscarinic receptors. PMID- 8405075 TI - Binding characteristics of remoxipride and its metabolites to dopamine D2 and D3 receptors. AB - The substituted benzamide, remoxipride, is a new atypical antipsychotic agent with good clinical efficacy and low extrapyramidal side-effect potential. In the present study, the in vitro receptor binding properties of remoxipride and several of its metabolites to rat striatal dopamine D2 and cloned human dopamine D2A and D3 receptors were investigated. Remoxipride bound to [3H]raclopride labelled dopamine D2 receptors in rat striatum with an affinity (Ki) of 113 nM. The significantly lower affinities of remoxipride reported when [3H]spiperone was used as a radioligand are suggested to be due to methodological problems associated with the use of very high-affinity radioligands. Some of the phenolic metabolites of remoxipride found mainly in rat exhibited considerably higher affinities to dopamine D2 and D3 receptors than remoxipride itself. The pyrrolidone metabolites found mainly in the human had very low dopamine D2 and D3 affinities. The present in vitro results suggest that the behavioural effects of remoxipride in rats may reflect the effect of remoxipride and some of its high affinity metabolites. PMID- 8405076 TI - Ceruletide suppresses rotational behavior in lesioned rats via CCKA receptors. AB - The effects and pharmacological mechanisms underlying the inhibiting effect of ceruletide, a cholecystokinin (CCK)-related peptide, on apomorphine-induced turning behavior in 6-hydroxydopamine lesioned rats were investigated. For this purpose, selective CCKA and CCKB receptor antagonists were used. Ceruletide (50 400 micrograms/kg, s.c.) dose dependently suppressed apomorphine-induced rotational behavior. The antiapomorphine effect of ceruletide was reversed by the selective CCKA receptor antagonist, devazepide, but not by the selective CCKB receptor antagonist, L-365,260. This suggests that the suppression ceruletide exerts on hyperactive nigrostriatal dopamine neurons is mediated by CCKA receptors. PMID- 8405077 TI - Nimodipine inhibits [3H]nitrobenzylthioinosine binding to the adenosine transporter in human brain. AB - The inhibition of [3H]nitrobenzylthioinosine ([3H]NBI) binding to human parietal cortex membranes by adenosine transport inhibitors, adenosine receptor agonists and antagonists and dihydropyridines was investigated. While the adenosine transport inhibitors inhibited [3H]NBI binding with Ki values in the low nanomolar range and the adenosine A1 receptor agonist, cyclopentyladenosine, with a Ki in the low micromolar range, no IC50 values could be obtained for the adenosine receptor antagonists at concentrations up to 100,000 nM. Among the dihydropyridines (+)-nimodipine was the most potent with a Ki of 201 +/- 55 nM. Inhibition of adenosine transport thus may contribute to the clinical effects of nimodipine in the central nervous system. PMID- 8405078 TI - Inhibition of hippocampal acetylcholine release by benzodiazepines: antagonism by flumazenil. AB - Diazepam (2.5-10 mg/kg i.p.) and midazolam (2.5-10 mg/kg i.p.) decreased acetylcholine release in the hippocampus of freely moving rats. This effect was antagonized by pretreatment with flumazenil (1 mg/kg i.p.). These results show that activation of benzodiazepine receptors reduces the in vivo release of acetylcholine in the hippocampus, suggesting that the septo-hippocampal cholinergic system, which has a major role in the regulation of cognitive functions, is under inhibitory control exerted by gamma-aminobutyrate (GABA) neurons. PMID- 8405079 TI - Characterisation of the peptido-leukotriene receptor PL2 on the ferret spleen strip. AB - The peptido-leukotriene receptor(s) (PL) on the ferret isolated spleen strip have been characterised by functional studies using the naturally occurring leukotrienes (LTs), a range of structurally distinct PL antagonists, and by ligand binding studies. LTB4 (0.01-10 microM) was inactive on ferret spleen whereas LTC4, LTD4 and LTE4 produced concentration-related contractions with maximal responses, relative to noradrenaline, of 57% (EC50 0.28 microM), 60% (EC50 0.5 microM) and 7% respectively. The leukotriene responses were unaltered by L-serine borate, L-cysteine, indomethacin, phentolamine, propranolol, mepyramine, methysergide or atropine, suggesting that the peptido-leukotrienes were acting through distinct PL receptors. The PL1 antagonists, FPL 55712 (0.01 10 microM), ICI 198615 (10 microM), SK&F 104353 (10 microM) and MK541 (10 microM) were all inactive against LTC4- or LTD4-induced contractile responses. LTE4 was a partial agonist with respect to LTC4 and LTD4 with pKB values of 5.8 and 5.5 respectively. Nifedipine (0.1 microM) produced a rightward shift of the concentration-response curves to both LTC4 and LTD4 and depressed their maximal responses. An unacceptably high level of non-specific binding of [3H]LTD4 to membrane preparations of ferret spleen prevented characterisation of this receptor by ligand binding. These results suggest that the ferret spleen has a homogeneous population of a PL receptor type which is insensitive to existing PL1 receptor antagonists. The functional characteristics of this PL receptor type are similar to those of the PL2 receptor on other tissues. The absence of PL1 receptors on this tissue makes it particularly useful in identifying new and selective drug tools for the PL2 receptor. PMID- 8405080 TI - Effects of (+)-S-12967 and (-)-S-12968, two enantiomers of a new slow-acting 1,4 dihydropyridine, on rat coronary resistance arteries. AB - The action of (+)-S-12967 and (-)-S-12968, two isomers of a new 1,4 dihydropyridine molecule (2-(-7-amino-2,5-dioxaheptyl)-3-ethoxycarbonyl-4-(2,3 dichlorop hen yl)-5-methoxycarbonyl-6-methyl 1,4-dihydropyridine), was studied on responses of rat isolated coronary resistance arteries (i.d. about 230 microns) to K+, Ca2+, and 5-hydroxytryptamine (5-HT). Both isomers slowly relaxed coronary arteries contracted with 125 mM K+, reaching a maximal effect in about 2 h. In contrast, the maximal relaxing effect of nifedipine was obtained within 20 min. The response to 125 mM K+ did not recover within the 2-h washout period in vessels exposed to the isomers but returned to pre-drug levels within 40 min in vessels exposed to nifedipine. Nifedipine was 4 times more potent than the (-) isomer which again was about 200 times more potent that the (+)-isomer. The IC50[M] values were approximately 1 nM, 4 nM and 0.8 microM, respectively. The relaxing effect of the isomers, which has a pKa of 8.6, was dependent on the extracellular pH being greater at high than low pH. Both isomers antagonized the vessel responses to K+ and Ca2+ and 5-HT. Higher concentrations of the isomers were required to antagonize responses to K+ and 5-HT than to Ca2+, probably due to the more depolarized state of the vascular smooth muscle in the latter experiments. In conclusion, the results demonstrate extracellular pH dependence as well as stereoselectivity regarding potency of (+)-S-12967 and (-)-S-12968 in rat coronary arteries.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405081 TI - Evaluation of truncated neuropeptide Y analogues with modifications of the tyrosine residue in position 1 on Y1, Y2 and Y3 receptor sub-types. AB - Substitutions of the tyrosine residue in position 1 of truncated neuropeptide Y (N-terminal fragment 1-4 linked to C-terminal fragment 18-36 by the epsilon aminocaproic acid) produced analogues that compete for specific [125I]polypeptide YY (PYY) binding in the frontoparietal cortex (Y1-enriched) with a profile best fitted to a two site-model with KD values in the low and high nM range, respectively. In the hippocampal membrane preparations (Y2-enriched), halogen substitutions on the aromatic ring generated analogues with competition profiles best fitted to a one-site model, revealing differences between the two binding assays and the interaction of these analogues with the Y1 and Y2 receptor sub types. In the rat vas deferens (Y2-enriched), all truncated analogues inhibited the twitch response with similar or slightly weaker potency than the native molecule. In contrast, these molecules were markedly less potent than neuropeptide Y (NPY) in the rabbit saphenous vein (Y1-enriched) and the rat distal colon (Y3-enriched). Some of the truncated analogues were inactive at up to microM concentrations in the rat distal colon, demonstrating the distinct structural requirement of the receptor sub-type present in this bioassay. These results revealed that amino acid residues between positions 5 and 17 are critical for the maintenance of optimal affinity for the NPY receptors present in the rabbit saphenous vein and the rat distal colon.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405082 TI - Role of endothelium-derived relaxing factor on coronary blood flow regulation in the dog. AB - The endothelium plays a key role in the regulation of vasoreactivity. To assess its importance on coronary flow regulation, we studied the participation of endothelium-derived relaxing factor-nitric oxide (EDRF-NO) on coronary reactive hyperemia and on the hyperemia that occurs secondary to an increase in myocardial oxygen consumption. In 15 dogs, the reactive hyperemic response decreased substantially after inhibition of EDRF-NO synthesis with N-omega-nitro-L-arginine (P < 0.01). In contrast, the hyperemia secondary to an increase in myocardial oxygen consumption, characterized by a linear correlation between myocardial oxygen consumption and coronary flow, did not change significantly after inhibition of EDRF-NO production (regression analysis, P > 0.1). Thus EDRF-NO synthesis by the endothelium is an important mechanism mediating the reactive hyperemic response but it does not seem to be essential for the metabolic regulation of coronary vascular resistance during hyperemia induced by an increased metabolic demand on the myocardium. PMID- 8405083 TI - The inhibitory effects of Ro 31-6930 and BRL 38227 on cholinergically-mediated bronchoconstriction in the guinea-pig. AB - This study compares the effects of two K(+0-channel openers, Ro 31-6930 and BRL 38227, on cholinergically-evoked contraction of guinea-pig airways to examine whether either compound acts through prejunctional inhibition of the release of acetylcholine. In the isolated trachea, Ro 31-6930 and BRL 38227 evoked concentration-dependent inhibition of tone generated by electrical field stimulation with pD2 values of 7.03 (6.77-7.29) and 6.26 (5.91-6.61) respectively and of that elicited by acetylcholine with pD2 values of 7.38 (6.52-8.24) and 6.65 (6.16-7.13). Neither compound was more potent against responses to electrical field stimulation than against acetylcholine. In the anaesthetised guinea-pig, Ro 31-6930 inhibited the bronchoconstriction evoked by bilateral vagus nerve stimulation and intravenous acetylcholine with ID50 values of 12.9 +/ 3.9 and 3.6 +/- 1.3 micrograms/kg i.v. respectively. The corresponding values for BRL 38227 were 356 +/- 157 and 37.9 +/- 13.4 micrograms/kg i.v. respectively. Thus, in vivo, both compounds were more potent against acetylcholine than against vagal stimulation. These results provide indirect evidence that K(+)-channel openers do not inhibit the release of acetylcholine from parasympathetic nerves in guinea-pig airway smooth muscle. PMID- 8405084 TI - Kinetic characterization of adenosine A2 receptor-mediated relaxation in isolated rabbit aorta. AB - Previous studies in our laboratory (Wiener et al., 1991, Soc. Neurosci. Abstr. 17, 989) have addressed aspects of the functional antagonism between the responses mediated by activated adenosine A2 receptors and alpha 1-adrenoceptors in adventitia- and endothelium-denuded rabbit thoracic aortic rings by steady state protocols which ignore the time course of response generation. In the present communication we describe aspects of the time-dependent kinetics of relaxation responses to adenosine A2 receptor agonists in tissues pre-contracted with the alpha 1-adrenoceptor agonist phenylephrine. The results were analyzed by application of the model originally developed by Keitz et al. (1990, J. Pharmacol. Exp. Ther. 255, 650) to describe the relaxation response, to a beta adrenoceptor agonist, as a first-order exponential decrease in tissue tension over time to estimate the apparent rate constant for relaxation (krel) and the magnitude of relaxation at equilibrium. The magnitude of the relaxation responses to adenosine, N6-cyclohexyladenosine, N6-methyladenosine, 5'-N ethylcarboxamidoadenosine, and R(-)-N6-(2-phenylisopropyl)adenosine were agonist concentration-dependent and saturable, as were the apparent rate constants for relaxation. In addition, the magnitude of the apparent rate constants for relaxation and the relaxation responses were inversely proportional to the fractional occupancy of the alpha 1-adrenoceptor. The hypothesis put forth by Keitz et al. that the maximal value of the apparent rate constant for relaxation may serve as the kinetic definition of agonist efficacy was also tested and found to be invalid for the adenosine A2 receptor. We propose that this pair of activated receptors and tissue preparation is a good model to study quantitative aspects of functional antagonism by kinetic paradigms. PMID- 8405085 TI - Smooth muscle relaxant effects of propofol and ketamine in isolated guinea-pig trachea. AB - The effects of anesthetics on airway smooth muscle tone are important in the management of patients with asthma. In the present study we evaluated the effect of propofol and ketamine on isolated guinea-pig tracheal preparations mounted for recording isometric contractile force. In a concentration-dependent way both drugs produced 100% relaxation irrespective of whether tracheal tone was spontaneous or induced by carbachol, histamine, prostaglandin F2 alpha, 30 mM K+ or 124 mM K+. The relaxant potency of propofol was dependent of the formulation of the drug used. Propofol showed an about 3 times higher potency when solubilized with hydroxypropyl-beta-cyclodextrin compared with an oil-in-water emulsion of the drug (Diprivan). Propofol had the greatest potency on tracheal preparations with spontaneous tone (EC50 = 4.0 +/- 0.9 microM). Ketamine preferentially relaxed contractions elicited by carbachol (EC50 = 120.8 +/- 5.2 microM) and had a lower potency than propofol when tone was spontaneous or induced by other tracheal spasmogens. Since propofol was a more effective tracheal relaxant in vitro than ketamine, the possibility that propofol, like ketamine, may inhibit bronchoconstriction during anesthesia should be studied further. PMID- 8405086 TI - Lisinopril increases the recovery during reoxygenation and resistance to oxidative damage in cardiomyocytes. AB - The action of the angiotensin-converting enzyme (ACE) inhibitor lisinopril on the consequences of myocardial reoxygenation and oxidative damage was assessed in cultured chick embryonic ventricular cardiomyocytes. Lisinopril, 10(-8) M to 10( 6) M, produced a significant (P < 0.05) dose-dependent enhancement of the restoration of contractile frequency occurring during myocardial reoxygenation but did not alter the depression in contractile frequency during hypoxia. Lisinopril significantly (P < 0.05) shifted the dose-response relationship of ammonium persulfate-induced reduction in cardiac contractile frequency. Lisinopril significantly (P < 0.05) reduced the effect of another oxidative agent, tertbutylhydroperoxide which produced a time-dependent reduction in cardiac contractile frequency. Lisinopril did not alter cardiac contractile frequency in the absence of hypoxia or ammonium persulfate or tertbutylhydroperoxide. The viability of cardiomyocytes, assessed by trypan blue exclusion, paralleled the changes in cardiac contractile frequency. Lisinopril significantly (P < 0.05) improved viability of cardiomyocytes exposed to either ammonium persulfate or tertbutylhydroperoxide. Lisinopril did not display any antioxidant properties against the free radical alpha,alpha-diphenyl-beta picrylhydrazyl. These data suggest that lisinopril accelerates the recovery of cardiomyocytes during reoxygenation and blunts the effects of oxidative agents through mechanisms involving the endogenous renin angiotensin system and/or a direct cellular action. PMID- 8405087 TI - Evidence of an exocytotic-like release of [3H]5-hydroxytryptamine induced by d fenfluramine in rat hippocampal synaptosomes. AB - The monoamine releasing activity of d-fenfluramine was investigated with an in vitro model consisting of synaptosomes preloaded with the 3H-neurotransmitter and extensively washed in a superfusion apparatus before a 3-min exposure to d fenfluramine. With this model, the drug-induced release is real and is not confused by inhibition of reuptake by the drug. d-Fenfluramine (0.5 microM) induced only [3H]5-hydroxytryptamine ([3H]5-HT) release from hippocampal synaptosomes whereas 10 microM also induced some overflow from hippocampal synaptosomes preloaded with [3H]noradrenaline or from striatal synaptosomes preloaded with [3H]dopamine, although the overflow was much lower than from 5 HTergic synaptosomes. We then focused on the [3H]5-HT release induced by 0.5 microM d-fenfluramine, which was previously shown to be Ca2+ dependent. The same finding was confirmed in the present study with other experimental protocols, indicating the requirement for extracellular Ca2+ ions. By measuring [3H]5-HT uptake into rat hippocampal synaptosomes we confirmed that Ca(2+)-ions are not required for the function of the 5-HT uptake carrier or for its interaction with d-fenfluramine. d-Fenfluramine-induced [3H]5-HT release was not altered by 1 microM nitrendipine (blocking the L-type Ca2+ channels) but was slightly decreased (20%) by 0.5 microM omega-conotoxin (blocking the N-type Ca2+ channels). It was also inhibited by 0.5 microM clonidine, interacting with alpha 2-adrenergic heteroreceptors, and by 10 nM tetanus toxin, known to affect the exocytosis of different neurotransmitters including 5-HT. These compounds had very similar effects on the Ca(2+)-dependent, exocytotic release of [3H]5-HT induced by depolarization, i.e. by 15 mM K+.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405088 TI - Gold ion induces contraction in frog skeletal muscle fibers. AB - In Ringer solution, gold ions (Au3+) at concentrations more than 50 microM produced a phasic and subsequent tonic contraction spontaneously in single toe muscle fiber of frog. When 1.8 mM Ca2+ in Ringer solution was replaced by 3 mM Mg2+, tonic contraction was no longer provoked in response to Au3+. Only phasic contraction was potentiated by 10 mM perchlorate (an L-type Ca2+ channel activator) irrespective of external Ca2+, and both phasic and tonic contractions were blocked by 10 microM nifedipine (an L-type Ca2+ channel blocker). Upon application of 5 mM dithiothreitol to the contracting fiber, the Au(3+)-induced tension disappeared rapidly. The fiber pretreated with 0.05% H2O2 for 10 min did not respond to Au3+ with visible contraction. Treatment of H2O2-paralyzed fibers with dithiothreitol (to reduce oxidized sulfhydryl groups) fully restored the Au(3+)-induced contraction. These results suggest that the phasic contraction induced by Au3+ probably is mediated through sulfhydryl groups in the L-type Ca2+ channel (dihydropyridine receptor) on the transverse tubular membrane. Sustained contraction was produced by Ca2+ application to Au(3+)-treated fibers in Mg(2+) Ringer solution, and Au3+ caused membrane depolarization in a dose-dependent manner. These effects of Au3+ may explain tonic contraction development. PMID- 8405089 TI - Protection from hypoxic and N-methyl-D-aspartate injury with azelastine, a leukotriene inhibitor. AB - The 5-lipoxygenase metabolites, leukotrienes, increase in concentration during cerebral ischemia. Azelastine is a new anti-allergic agent which inhibits leukotriene C4 synthesis and release. We examined the neuroprotective properties of azelastine using the hippocampal slice. Azelastine 15 microM significantly protected CA1 evoked responses from hypoxic injury, with CA1 population spike amplitude recovering to a mean 76 +/- 13% in azelastine treated slices, compared to 4 +/- 3% recovery in paired unmedicated slices. The EC50 for this azelastine hypoxic protection was 9.8 microM. Azelastine additionally protected against injury induced by N-methyl-D-aspartate (NMDA), but not non-NMDA glutamate receptor agonists. No hypoxic protection was afforded by diphenhydramine 50 microM, suggesting that azelastine protection did not occur through histamine H1 receptor blockade. The finding of protection with azelastine against hypoxic and NMDA-induced injury suggests that leukotriene production is a common pathway in these forms of neuronal injury, and that leukotriene inhibition may be a useful neuroprotective strategy. PMID- 8405090 TI - Multiple mechanisms of serotonin 5-HT2 receptor desensitization. AB - Desensitization of serotonin 5-HT2 receptor-mediated enhancement of the N-methyl D-aspartate (NMDA) depolarization was studied in rat cortical neurons. Serotonin and (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI) induced long term desensitization. Staurosporine, a nonspecific protein kinase C inhibitor, potentiated the serotonin and DOI facilitation, suggesting acute desensitization was operative. In the case of DOI, long term desensitization was prevented by staurosporine. Activators of protein kinase C abolished the serotonin facilitation, an action prevented by staurosporine. Concanavalin A potentiated the facilitation at 100 microM, but not 30 microM serotonin, suggesting these receptors undergo dose dependent internalization. Calmodulin antagonists prevent long term desensitization induced by serotonin. The depolarization induced by NMDA alone was not altered by staurosporine, protein kinase C activators, concanavalin A or calmodulin antagonists. Serotonin at 100 microM, but not 30 microM, induced heterologous desensitization of phenylephrine and carbachol induced facilitation of the NMDA depolarization. We conclude that serotonin 5-HT2 receptors both induce and undergo several forms of desensitization. PMID- 8405091 TI - Effects of sustained release formulation of thyrotropin-releasing hormone on learning impairments caused by scopolamine and AF64A in rodents. AB - The effects of a sustained-release formulation of thyrotropin-releasing hormone (TRH-SR) on learning impairments induced by scopolamine and a cholinergic neurotoxin, ethylcholine aziridinium ion (AF64A), were examined in rodents. Subcutaneous injection of TRH-SR (2.8 mg/kg as free TRH) produced a sustained increase in immunoreactive plasma TRH levels up to about 2 weeks after dosing in rats. TRH-SR (0.56 and 2.8 mg/kg) given subcutaneously 7 days before the acquisition trial markedly ameliorated scopolamine-induced amnesia in mice, as evaluated with a passive avoidance task. Repeated administration of TRH for 7 days at doses of 0.2-5 mg/kg s.c. elicited a dose-dependent recovery from amnesia induced by scopolamine, whereas only the group treated with 5 mg/kg/day showed a significant improvement. The rats with bilateral intracerebroventricular injection of AF64A (3.75 nmol/brain) showed a significant impairment in the water maze task 2 weeks after surgery. TRH-SR (0.56 and 2.8 mg/kg) also exhibited a dose-dependent ameliorating action on the deficit. These findings indicate that TRH-SR ameliorates learning impairments produced by scopolamine and AF64A, and suggest that continuous infusion of TRH may have a potent learning and memory improving action at low doses. PMID- 8405092 TI - Differences in anticonvulsant potency and adverse effects between dextromethorphan and dextrorphan in amygdala-kindled and non-kindled rats. AB - The anticonvulsant and adverse effects of dextromethorphan, a non-opioid antitussive, and its metabolite dextrorphan were examined in amygdala-kindled rats. Both drugs have repeatedly been proposed to be functional non-competitive N methyl-D-aspartate (NMDA) receptor antagonists, but they also exert effects distinct from antagonism at NMDA receptors, such as blockade of voltage-gated calcium channels and sigma-site mediated actions. Since recent data have demonstrated that kindled rats are more susceptible to the adverse effects of NMDA receptor antagonists than non-kindled rats, the time course, characteristics and severity of adverse effects of dextromethorphan and dextrorphan were also determined in non-kindled animals. Dextromethorphan dose dependently increased the focal seizure threshold (i.e. the threshold for induction of afterdischarges recorded from the amygdala) in fully kindled rats. This anticonvulsant effect was found at relatively low doses (7.5-15 mg/kg i.p.) which were almost free of any adverse effects. At higher doses, dextromethorphan induced motor impairment and seizures, but no phenyclidine (PCP)-like adverse effects, such as hyperlocomotion or stereotypies. In contrast, such adverse effects were seen after dextrorphan, although only infrequently. Dextrorphan was less potent in inducing anticonvulsant but more potent in inducing motor impairing effects than dextromethorphan in kindled rats. In non-kindled rats, the motor impairment induced by dextrorphan was significantly less severe than in kindled rats, whereas no marked differences between kindled and non-kindled rats were found for dextromethorphan. The data indicate that dextromethorphan and dextrorphan differ in their mechanisms of action. Only dextrorphan exerts effects which are characteristic for NMDA receptor antagonism, whereas the potent anticonvulsant effect of dextromethorphan in presumably unrelated to the NMDA receptor complex. PMID- 8405093 TI - The leukotriene D4 receptor blocker, SK&F 104353, inhibits volume regulation in isolated crypts from the rat distal colon. AB - The effect of SK&F 104353 (2-hydroxy-3-carboxyethylthio-3-[2-(8 phenyloctyl)phenyl] propanoic acid), a specific leukotriene D4 receptor blocker, on volume regulation in isolated rat colonic crypts was studied. SK&F 104353 (10( 7)-5 x 10(-6) mol.l-1) concentration dependently inhibited the regulatory decrease in volume following cell swelling induced either by uptake of a short chain fatty acid, butyrate, or by exposure to hypotonic medium. Whole-cell patch clamp experiments revealed that SK&F 104353 suppressed the depolarization of crypt cells during the regulatory decrease in volume. Also the effect of leukotriene D4 (5 x 10(-7) mol.l-1), which has an action on the membrane potential similar to that induced by cell swelling, was suppressed nearly completely by the receptor blocker. In contrast, the precursor of leukotriene D4, leukotriene C4, had no effect on the membrane potential. These results are in accordance with the hypothesis that leukotriene D4 acts as mediator for the activation of basolateral Cl- channels in the rat colonic epithelium during regulatory decrease in volume. PMID- 8405094 TI - Cardiorespiratory effects produced by blockade of excitatory amino acid receptors in cats. AB - The aim of our study was to determine the role of excitatory amino acids in controlling cardiorespiratory activity. For this purpose we administered an antagonist of both N-methyl-D-aspartate (NMDA) and non-NMDA receptors (kynurenic acid), and an antagonist of the NMDA receptor complex (dizocilpine, more commonly known as MK-801) i.v. to chloralose-anesthetized cats while monitoring tracheal air flow, tidal volume, respiratory rate, inspiratory and expiratory durations, end tidal CO2, arterial blood pressure and heart rate. Administration of kynurenic acid in doses of 350 and 500 mg/kg produced respiratory depression as reflected by decreases in respiratory minute volume and increases in end tidal CO2. Inspiratory duration was increased with both doses and apnea (occurring during expiration) was observed with the high dose. Apnea was preceded by an apneustic pattern of breathing. Both doses resulted in an increase in blood pressure and, with the high dose, a later decrease in blood pressure was noted. Dizocilpine in doses ranging from 0.03 to 1 mg/kg produced dose-related decreases in respiratory minute volume, and increases in end tidal CO2. In addition, dizocilpine produced increases in inspiratory duration, an apneustic pattern of breathing and apnea (occurring during inspiration). Effects on blood pressure were similar to those observed with kynurenic acid. It is concluded that blockade of excitatory amino acid receptors results in pronounced effects on cardiorespiratory activity. PMID- 8405095 TI - Effects of indirectly acting 5-HT receptor agonists on circulating melatonin levels in rats. AB - Because circulating melatonin levels are generally thought to be under the strict control of pineal N-acetyltransferase, little attention has been paid to the impact of an altered availability of serotonin (5-HT) on melatonin formation. In order to see whether melatonin synthesis is stimulated by an increased availability of free, cytosolic 5-HT, we studied the effects of 5-HT precursors, 5-HT releasers and reuptake inhibitors and of monoamine oxidase inhibitors, alone and in combination, on circulating melatonin levels in experimental animals. The administration of tryptophan and 5-HT-releasing drugs (fenfluramine, +/- 3,4 methylenedioxymethamphetamine) to rats caused a dose- and time-dependent elevation of circulating melatonin levels during the day and night. This increase in melatonin was further enhanced by inhibition of monoamine oxidase. The elevation of plasma melatonin caused by 5-HT-releasing drugs was prevented by prior administration of fluoxetine. Monoamine oxidase inhibitors and fluoxetine alone had no effect on circulating melatonin levels. These findings indicate that the administration of indirectly acting 5-HT receptor agonists which increase the free cytoplasmic pool of 5-HT may also elevate circulating melatonin levels. The results of this study suggest that the rate of pineal melatonin synthesis is dependent on the free cytoplasmic pool of 5-HT in pinealocytes and that the drug induced elevation of this pool stimulates melatonin formation and increases circulating melatonin levels. At least some of the effects of indirectly acting 5 HT receptor agonists, e.g. on sleep, mood, food intake, pain perception, and neuroendocrine secretion, may therefore be mediated by the elevation of circulating melatonin and the subsequent activation of central melatonin receptors. PMID- 8405096 TI - Stress activation of mesocorticolimbic dopamine neurons: effects of a glycine/NMDA receptor antagonist. AB - Restraint of brief duration causes a metabolic activation of mesocortical and some mesolimbic dopaminergic systems with little effect on the nigrostriatal system. We have examined the ability of an antagonist of the allosteric glycine site of the N-methyl-D-aspartate receptor complex to block the stress-induced response in dopamine utilization. Thirty minutes of restraint stress elevated dopamine metabolism, as measured by the ratio between 3,4-dihydroxyphenylacetic acid (DOPAC) and dopamine, in both the medial prefrontal cortex and nucleus accumbens. An antagonist for the glycine/N-methyl-D-aspartate receptor complex, 1 hydroxy-3-aminopyrrolidone-2 ((+)-HA-966), given systemically or injected into the ventral tegmental area, prevents the stress-induced increase in dopamine metabolism in the prefrontal cortex without altering the response in the nucleus accumbens. Similarly, systemic administration of the non-competitive antagonist for the N-methyl-D-aspartate receptor, dizocilpine ((+)-MK-801), blocked the stress-induced rise in dopamine metabolism in the medial prefrontal cortex but not the nucleus accumbens. The negative enantiomer of HA-966 did not produce a selective antagonism of the stress-induced dopamine metabolism in the medial prefrontal cortex. These results support previous work which suggest the mesocortical and mesoaccumbens dopamine neurons respond to excitatory input through different glutamate receptor mechanisms. Additionally, the specific blockade of the stress-induced change in dopamine metabolism in the medial prefrontal cortex by a glycine antagonist implies a role for such an antagonist in treatment of disease states which may involve disruptions of N-methyl-D aspartate receptor function. PMID- 8405097 TI - Brief cortisol exposure elevates adrenal phenylethanolamine N-methyltransferase after a necessary lag period. AB - The present study, using bovine adrenal medullary cells, characterized in detail the time course of regulation of phenylethanolamine N-methyltransferase activity following brief glucocorticoid exposure. Cortisol pulses (10(-4) and 10(-5) M), as short as 15 min, increased phenylethanolamine N-methyltransferase activity measured 2 days following cortisol exposure, with a required lag period of 18 h or more. Phenylethanolamine N-methyltransferase activity was increased 2 days following brief (2 h) exposure to cortisol in concentrations that reach the medulla in vivo (10(-6) to 10(-4) M). Phenylethanolamine N-methyltransferase activity following both continuous and 2 h pulses of 10(-5) M cortisol were reduced by the glucocorticoid receptor antagonist, RU 38486. A 2 h pulse of nicotine (10(-5) M) increased phenylethanolamine N-methyltransferase activity with a lag period of at least 18 h, while combination treatment of nicotine and cortisol (10(-4) M) produced significantly higher increases in phenylethanolamine N-methyltransferase compared to either treatment alone. Therefore, this study provides novel in vitro evidence for the regulation of adrenomedullary phenylethanolamine N-methyltransferase activity, following a necessary lag period, by acute changes in both cortisol and nicotine. PMID- 8405098 TI - DM-9384, a new cognition-enhancing agent, increases the turnover of components of the GABAergic system in the rat cerebral cortex. AB - DM-9384 (nefiracetam) (N-(2,6-dimethylphenyl)-2-(2-oxo-1-pyrrolidinyl)acetamide), a pyrrolidone derivative (or a cyclic derivative of gamma-aminobutyric acid (GABA)), is a newly developed nootropic (or cognition-enhancing) agent. In the present study, we examined the biochemical effect of DM-9384 on GABAergic neurons in adult rat brains. DM-9384, when administered orally at a daily dose of 10 mg/kg for 7 days, significantly increased GABA turnover and glutamic acid decarboxylase activity in the cortex and hippocampus, and stimulated Na(+) dependent high-affinity GABA uptake in cortical synaptosomes. In in vitro experiments, the K(+)-evoked release of [14C]GABA from cortical slices was markedly increased by low concentrations (10(-8), 10(-9) M) of DM-9384. The binding of GABAA and benzodiazepine to their receptors in the brain was not affected by DM-9384 (10(-10)-10(-3) M). The results suggest that DM-9384 increases the turnover of components of the GABAergic system by influencing presynaptic sites rather than postsynaptic sites. PMID- 8405099 TI - Capsazepine-sensitive release of calcitonin gene-related peptide from C-fibre afferents in the guinea-pig heart by low pH and lactic acid. AB - The present study aimed to evaluate the possible influence of the selective capsaicin antagonist, capsazepine, on the release of calcitonin gene-related peptide (CGRP)-like immunoreactivity from sensory nerves in the isolated perfused guinea-pig heart. Low-pH buffer (pH 7, 6, 5), capsaicin (10(-7) M), lactic acid (5, 20, 50 mM) and nicotine (10(-4) M) all evoked a clear-cut release of CGRP like immunoreactivity. Incubation with capsazepine (10(-6) to 10(-5) M) significantly reduced the CGRP-like immunoreactivity release evoked by low pH, capsaicin and lactic acid (5 mM) but not that evoked by nicotine. Furthermore, the capsaicin-evoked stimulation of heart rate was inhibited by incubation with capsazepine. The inorganic dye, ruthenium red, which has previously been shown to attenuate capsaicin-, but not nicotine-induced CGRP release from the heart, also reduced the release of CGRP caused by low pH and lactic acid (5 mM). It is concluded that the CGRP-like immunoreactivity release evoked from the heart by low pH and lactic acid shares several characteristic features with the release evoked by capsaicin. Since tissue pH is low in myocardial ischaemia and this is well known to cause pain, the use of capsazepine to inhibit the function of C fibre afferents may represent a novel principle to influence autonomic reflex reactions associated with cardiac pathological conditions. PMID- 8405100 TI - Loop and distal actions of a novel diuretic, M17055. AB - We investigated the mechanism of action of a novel 'high ceiling' diuretic, M17055, in in vivo clearance studies with anesthetized dogs during water diuresis and in vitro microperfusion studies of isolated rabbit renal tubules. In the clearance study, intravenous infusion of M17055 (1 mg/kg per h) decreased free water clearance and increased urinary excretion of Na+ and Cl- to a greater extent than did a maximum dose of furosemide (30 mg/kg per h). With the maximum dose of furosemide, an additional dose of M17055 or hydrochlorothiazide resulted in additional suppression of free water clearance. These results indicate that M17055 has some additional mechanisms of action in the distal nephron. In isolated rabbit cortical thick ascending limb of Henle's loop, M17055 applied to the lumen decreased the lumen positive transepithelial voltage at concentrations over 10(-6) M and suppressed the lumen-to-bath 36Cl- flux at 10(-5) M. In the connecting tubule, M17055 added to the lumen suppressed lumen negative transepithelial voltage in a concentration-dependent manner in a range from 10( 4) to 10(-3) M. The effect of M17055 on transepithelial voltage was also observed in the distal convoluted tubule and cortical collecting duct. Moreover, 10(-3) M of M17055 in the lumen significantly decreased the lumen-to-bath 22Na+ flux in the cortical collecting duct. From these observations, it appears that M17055 acts not only on the thick ascending limb of Henle's loop but also on the distal segments via inhibition of electrogenic Na+ transport. PMID- 8405101 TI - Defibrotide, a single-stranded polydeoxyribonucleotide acting as an adenosine receptor agonist. AB - The binding of single-stranded polydeoxyribonucleotides to adenosine A1 and A2 receptors was investigated. Defibrotide, a natural substance with established anti-thrombotic and anti-ischaemic effects, displaced [3H]CHA (N6-cyclohexyl adenosine) and [3H]NECA (5'-N-ethylcarboxamido-adenosine) concentration dependently, completely and competitively. Ki values of 371 +/- 68 and 688 +/- 115 micrograms/ml (mean +/- S.E.M. of 4-5 replications) were computed for adenosine A1 and A2 sites, respectively. Higher and lower molecular weight polydeoxyribonucleotides displayed comparable affinity, whereas a double-stranded polydeoxyribonucleotide and a polyanion with a negative charge comparable to that of defibrotide were inactive. Defibrotide did not affect the total number of binding sites in radioligand saturation experiments. Defibrotide relaxed the K(+) contracted guinea-pig trachealis muscle (IC50 = 4001 micrograms/ml) about one third as potently as the CHA-contracted preparation and as potently as the resting preparation. NECA, a mixed adenosine A1/A2 receptor agonist, behaved similarly. The effects were abolished by the adenosine A1/A2 receptor blocker 8 phenyltheophylline, but not by the selective A1 blocker, 1,3-dipropyl-8-(2-amino 4-chlorophenyl)-xanthine. These results demonstrate that defibrotide binds to adenosine receptors and triggers pharmacological responses comparable to those of a known agonist. PMID- 8405102 TI - Inhibition of 9 alpha,11 beta-prostaglandin F2-induced bronchial hyperresponsiveness by thromboxane A2 receptor antagonists in guinea pigs. AB - We studied the effect of intravenous administration of 9 alpha,11 beta prostaglandin F2 on bronchial responsiveness to histamine and airway wall thickening in guinea pigs. The infusion of 9 alpha,11 beta-prostaglandin F2 induced an increase of the relative thickness of the airway wall in peripheral bronchi demonstrable by histological examination. Analysis of airway function showed that the infusion of 9 alpha,11 beta-prostaglandin F2 induced airway hyperresponsiveness to histamine with airway wall thickening. The thromboxane A2 receptor antagonists, ONO-NT-126 (5(Z)-6-[(1R,2R,3R,4S)-3-(n-4 bromobenzenesulfonyl-aminomethyl) bicyclo[2.2.1]heptane-2-yl]hex-5-enoic acid) and ONO-8809 (n-decyl(Z)-6-[(1S,2S,3R,4R)-3-(4-bromobenzenesulfonylaminomethyl) bicyclo[2.2.1]hept-2-yl]-5-hexenoate), inhibited these effects of 9 alpha,11 beta prostaglandin F2 in a dose-dependent manner. PMID- 8405103 TI - Characterization of muscarinic receptors mediating vasodilation in rat perfused kidney. AB - The muscarinic receptor mediating vasodilation of resistance vessels in the rat isolated, constant-pressure perfused kidney (preconstriction by 10(-7) M cirazoline) was characterized by subtype-preferring agonists and selective antagonists. The agonists produced vasodilation with the following rank order of potency: arecaidine propargyl ester (APE) > 5-methylfurtrethonium = methacholine = oxotremorine > (S)-aceclidine > arecaidine 2-butyne-1,4-diyl bisester > 4-Cl McN-A-343 = (R)-nipecotic acid ethyl ester = N-ethyl-guvacine propargyl ester approximately (R)-aceclidine = (S)-nipecotic acid ethyl ester > McN-A-343. Agonist-induced vasodilation disappeared after destruction of the endothelium with detergent. Highly significant correlations of agonist potencies for vasodilation were found between rat kidney and guinea-pig ileum submucosal arterioles as well as agonist potencies at smooth muscle muscarinic M3 receptors of the guinea-pig ileum. The rank order of antagonist potencies (4 diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) > (R)-hexahydro-difenidol approximately hexahydro-sila-difenidol > pirenzepine approximately p-fluoro hexahydro-sila-difenidol approximately himbacine approximately AF-DX 384 approximately AQ-RA 741 > (S)-hexahydro-difenidol) to attenuate vasodilation to APE in rat kidney, correlated significantly with affinities at M3 receptors in submucosal arterioles and in smooth muscle of the guinea-pig ileum, but differed from those at M1 and M2 receptors in rabbit vas deferens. The agonist and antagonist potencies suggest that vasodilation elicited by muscarinic stimuli in endothelium-intact rat renal vasculature is mediated by functional muscarinic M3 receptors. PMID- 8405104 TI - Mechanisms of cisplatin- and m-chlorophenylbiguanide-induced emesis in ferrets. AB - We investigated the involvement of peripheral and central serotonin (5-HT)3 receptors in cisplatin- and 5-HT3 receptor agonist-induced emesis in ferrets. Cisplatin (10 mg/kg i.v.)-induced emesis was inhibited by intravenous YM060 (0.003-0.1 microgram/kg). A highly selective and potent 5-HT3 receptor agonist, m chlorophenylbiguanide (1-10 mg/kg i.p.), dose dependently elicited emesis an effect which was inhibited by YM060 (0.003-0.3 microgram/kg i.v.). Vagotomy markedly reduced this emesis, and the combination of abdominal vagotomy and greater splanchnicectomy abolished emesis. Lesion of greater splanchnic nerves alone did not markedly inhibit emesis. Intracerebroventricularly (4th ventricle) administered YM060 inhibited cisplatin- and m-chlorophenylbiguanide-induced emesis only at higher doses (0.01-0.1 and 0.01-0.03 microgram, respectively). Intracerebroventricularly (4th ventricle) administered m-chlorophenylbiguanide (30-100 micrograms) produced only a weak retching response. These results indicate that stimulation of abdominal vagal afferent nerves via peripheral 5-HT3 receptors is important for triggering cisplatin- and m-chlorophenylbiguanide induced emesis in ferrets. PMID- 8405105 TI - Effects of cocaine and amphetamine on acetylcholine release in the hippocampus and caudate nucleus. AB - The role of dopamine in the control of hippocampal acetylcholine release was evaluated by using in vivo microdialysis. The effects of the two psychostimulants, cocaine and d-amphetamine, were studied on acetylcholine release in the hippocampus and compared to effects observed in the caudate nucleus. Administration of cocaine (10 and 20 mg/kg i.p.) increased acetylcholine release by 130 and 190% in the hippocampus, whereas in the caudate nucleus the enhancement was 51 and 80% over basal values, respectively. After the injection of d-amphetamine (1 and 2 mg/kg i.p.) the enhancement of acetylcholine release was 110 and 210% in the hippocampus whereas it was 35 and 54%, respectively, in the caudate nucleus. As observed in the caudate nucleus, pretreatment with the dopamine D1 receptor antagonist, SCH 23390, antagonized the cocaine- and amphetamine-induced increase in hippocampal acetylcholine release. These results show that cocaine and d-amphetamine, by increasing dopaminergic transmission, enhance the extracellular concentrations of acetylcholine in both brain areas. The relative enhancement in the hippocampus was far greater than that in the caudate nucleus, suggesting that dopaminergic control of cholinergic function differs in these two brain areas. The results also suggest that endogenous dopamine, by facilitating the release of acetylcholine in the hippocampus, may participate in the regulation of hippocampal cognitive processes. PMID- 8405106 TI - Defective stimulation of cyclic AMP by prostaglandin E2 in colonic epithelial cells in colitis. AB - The present study investigated the effect of vasoactive intestinal peptide and prostaglandin E2 on cyclic adenosine monophosphate levels in isolated colonocytes during experimental colitis. Intra-rectal trinitrobenzenesulfonic acid induced a colitis-like inflammation in the rabbit distal colon. Basal levels of cyclic adenosine monophosphate were similar in control and colitic colonocytes. Levels were increased by prostaglandin E2 and vasoactive intestinal peptide in control cells. Colonocytes from colitic tissue responded to vasoactive intestinal peptide normally, but exhibited an attenuated response to prostaglandin E2. We conclude during colitis the epithelium exhibits a specific alteration in prostaglandin E2 receptor number, affinity or adenylate cyclase coupling. PMID- 8405107 TI - Dexamethasone inhibits the pyrogenic activity of prostaglandin F2 alpha, but not prostaglandin E2. AB - The effect of dexamethasone on prostaglandin (PG) E2- and PGF2 alpha-induced fever was studied in rats. Intracerebroventricular injection of PGE2 and PGF2 alpha (500 ng) induced increases in body temperature (maximal temperature rises of 0.97 +/- 0.13 degrees C and 0.78 +/- 0.18 degrees C, respectively, vs. vehicle 0.12 +/- 0.09 degrees C) of unrestrained rats maintained within the thermoneutral zone. PGE2-induced fever peaked earlier and the defervescence was faster when compared to the response induced by PGF2 alpha. Subcutaneous pre-administration of dexamethasone (0.5 mg/kg) did not affect PGE2-induced fever (maximal temperature rise of 1.00 +/- 0.08 degrees C), but completely prevented the pyrogenic activity of PGF2 alpha (maximal temperature rise of 0.16 +/- 0.16 degrees C). Neither PGE2- nor PGF2 alpha-induced fever was significantly altered (maximal temperature rises of 0.90 +/- 0.11 degrees C and 0.64 +/- 0.14 degrees C, respectively) by intraperitoneal administration of indomethacin (2 mg/kg). These results demonstrate for the first time that glucocorticoids, in addition to inhibiting endotoxin- and cytokine-induced fever, can also modulate the pyrogenic activity of some prostaglandins, possibly via suppression of the synthesis of corticotropin-releasing factor, indicating that multiple mechanisms may be involved in the antipyretic activity of these steroids. PMID- 8405108 TI - Inhibitory action of N omega-nitro-L-arginine methyl ester on in vivo long-term potentiation in the rat dentate gyrus. AB - Recent in vitro observations have led to the suggestion that nitric oxide (NO) plays a modulatory role in the expression of long-term potentiation (LTP). We investigated whether an NO synthesis inhibitor, N omega-nitro-L-arginine methyl ester (L-NAME), affects the generation of LTP in anesthetized rats in vivo. Administration of L-NAME (0.1 and 1 nmol, i.c.v.) suppressed the magnitude of LTP dose dependently in the dentate gyrus of anesthetized rats. Co-injection of L arginine, which interferes with the inhibitory action of L-NAME on NO synthesis, reversed these effects, whereas there was no reversal in rats that received co administration with D-arginine. These results provide further support for the hypothesis that NO plays a modulatory role in the expression of synaptic potentiation. PMID- 8405109 TI - The regeneration of d,l-fenfluramine-destroyed serotonergic nerve terminals. AB - The regeneration of serotonergic nerve terminals subsequent to their destruction by high-dose fenfluramine administration was examined. Treating rats with fenfluramine (80 mg/kg over 2 days) destroyed 80% of serotonergic nerve terminals, indicated by reduced maximal [3H]paroxetine binding to 5 hydroxytryptamine (5-HT) uptake sites on synaptic membranes (Bmax) and maximal [14C]5-HT uptake rate into synaptosomes (Vmax). 25 weeks later, these indices of serotonergic nerve terminals had returned to 72% of control. Maximal synaptosomal loading (alpha) with [14C]5-HT also recovered (to 79% of control), reflecting an increased number of serotonergic synaptosomes. This suggests that the rebound in 5-HT uptake site density found after fenfluramine illustrates the regeneration of 5-HT-containing nerve endings. PMID- 8405110 TI - Evidence for separate Na+, K+, Cl- and K+, Cl- co-transport systems in mouse pancreatic beta-cells. AB - The effects of H 25 and H 74 on ouabain-resistant 86Rb+ influx were tested in beta-cell-rich mouse pancreatic islets. Both H 25, which is considered to be a specific inhibitor of Na+, K+, Cl- co-transport, and H 74, a specific inhibitor of K+, Cl- co-transport, reduced the ouabain-resistant 86Rb+ influx. The specific inhibitory effects of H 25 and H 74 on 86Rb+ influx did not overlap. The data suggest that mouse pancreatic beta-cells are equipped with separate systems for Na+, K+, Cl- and K+, Cl- co-transport. PMID- 8405111 TI - The effects of dorsal raphe administration of eltoprazine, TFMPP and 8-OH-DPAT on resident intruder aggression in the rat. AB - To evaluate the role of somatodendritic 5-HT1A receptors in the mediation of aggressive behaviour, eltoprazine, TFMPP (1-(3-trifluoromethylphenyl)piperazine hydrochloride) and 8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) were administered locally into the dorsal raphe nucleus of rats. 8-OH-DPAT (1 and 10 micrograms) and eltoprazine (10 and 30 micrograms) reduced aggression, but concomitantly reduced social interest and increased inactivity. TFMPP (1 and 10 micrograms) did not reduce aggression. As 8-OH-DPAT and to a lesser extent eltoprazine affect 5-HT1A receptors, it is proposed that a general reduction of serotonergic neurotransmission by activation of somatodendritic serotonergic autoreceptor leads to a non-specific reduction of aggression. As TFMPP has a significantly lower affinity for 5-HT1A receptors than 8-OH-DPAT or eltoprazine, the lack of effect of TFMPP supports this view. PMID- 8405112 TI - Alpha-atrial natriuretic peptide attenuates ethanol withdrawal symptoms. AB - Mice were rendered tolerant to and dependent on ethanol with an ethanol-liquid diet for 14 days. Five hours after withdrawal from ethanol, withdrawal symptoms were analyzed by scoring handling-induced convulsions. Intracerebroventricular (i.c.v.) injection of alpha-atrial natriuretic peptide (alpha-ANP) attenuated, whereas that of an antiserum against alpha-ANP (anti-ANP) intensified the severity of handling-induced convulsions. PMID- 8405113 TI - Repeated cocaine administration does not affect 5-HT receptor subtypes (5-HT1A, 5 HT2) in several rat brain regions. AB - In order to examine whether cocaine-induced behavioral sensitization is modulated by changes in serotonin receptor subtypes, we measured the binding of [3H]8 hydroxy-2-(di-n-propylamino)tetralin ([3H]8-OH-DPAT) to 5-HT1A receptors and of [3H]-ketanserin to 5-HT2 receptors in various brain regions of cocaine-treated and saline-treated (control) rats. As previously reported, repeated administration of cocaine resulted in behavioral sensitization. Stereotypic scores with the cocaine challenge were significantly (P < 0.05) higher in cocaine pretreated animals than in the saline-pretreated group. Neither acute nor chronic cocaine administration significantly altered the number (Bmax) or the affinity (KD) of either [3H]8-OH-DPAT or [3H]ketanserin binding sites in any of the brain regions examined. These results suggest that the enhanced functional sensitivity of 5-HT1A or 5-HT2 receptor subtypes seen with cocaine may be associated with alterations in processes distal to receptors rather than changes in the number or the affinity of the receptors. PMID- 8405114 TI - Tetrandrine: a vasodilator of medicinal herb origin with a novel contractile effect on dog saphenous vein. AB - During our recent pharmacological characterization of the vasodilator effects of tetrandrine on endothelium-denuded rings of dog saphenous vein, we have unexpectedly observed a slowly developing transient contractile response upon washout of KCl following the relaxation by tetrandrine of KCl-induced contraction. This washout-induced contraction in the presence of tetrandrine was most prominent in dog saphenous vein, smaller in dog mesenteric vein and not observed in dog mesenteric artery, dog aorta and rat aorta. This transient contraction induced in the presence of tetrandrine was not affected by atropine, indomethacin or prazosin, but was substantially inhibited by phentolamine and rauwolscine. Addition of amiloride or readmission of KCl inhibited the transient contraction upon washout of high KCl in the presence of tetrandrine, whereas addition of ouabain turned this transient contraction into a sustained one. Our results suggest that tetrandrine, in addition to its well known vasodilator effect via the blockade of Ca2+ channels, elicited a novel contractile effect which is specifically associated with the activation of postjunctional alpha 2 adrenoceptors functionally characteristic in venous smooth muscle. PMID- 8405115 TI - Mechanism of inhibitory action of ethanol on endothelium-dependent relaxation in rat aorta. AB - Using isolated rat aortic strips, we investigated the inhibitory effect of ethanol on endothelium-dependent relaxation induced by acetylcholine, especially on that mediated by endothelium-derived relaxing factor. Ethanol depressed the relaxation induced by acetylcholine and inhibited the increase in the content of intravascular cyclic GMP induced by acetylcholine, but not that induced by sodium nitroprusside or calcimycin. Ethanol also inhibited the acetylcholine-induced relaxation resistant to nitro-L-arginine. These results suggest that ethanol can inhibit the cyclic GMP-dependent relaxation mediated by endothelium-derived relaxing factor. Furthermore, ethanol seems to depress the cyclic GMP-independent relaxation mechanism. PMID- 8405116 TI - (+)-HA 966, a partial agonist at the glycine site coupled to NMDA receptors, blocks formalin-induced pain in mice. AB - (+)-(1-Hydroxy-3-aminopyrrolidine-2-one) ((+)-HA 966), a partial agonist at the glycine site coupled to N-methyl-D-aspartic acid (NMDA) receptors, abolished the late phase of licking induced by injection of formalin into the hind-paw of mice; inhibitory dose50 (ID50) = 1.6 mg/kg, s.c. In contrast, it was weakly active against the first phase; ID50 = 33.3 mg/kg, s.c. Further, (+)-HA 966 was inactive in the rotarod test of ataxia. These data support a role of NMDA receptors in the transmission of prolonged noxious stimulation and suggest that partial glycine receptor agonists may exert antinociceptive properties against persistent pain. PMID- 8405117 TI - Effects of muscarinic receptor agonists and anticholinesterase drugs on high voltage spindles and slow waves. AB - The effects of muscarinic agonists (AF102B, pilocarpine, oxotremorine) and anticholinesterases (physostigmine, tetrahydroaminoacridine) were investigated on the incidence of thalamically generated rhythmic high voltage spindles and on scopolamine (0.2 mg/kg)-induced neocortical slow wave activity (i.e. increased sum amplitude value of the 1-20 Hz band in a quantitative electroencephalography (qEEG) analysis). AF102B and pilocarpine decreased high voltage spindles and scopolamine increased sum amplitude values at 3 and 9 mg/kg, but not at 1 mg/kg. Oxotremorine was less potent than AF102B or pilocarpine in suppressing high voltage spindles. Oxotremorine had no effect on the scopolamine-induced qEEG changes. Tetrahydroaminoacridine decreased high voltage spindles at 1, 3 and 9 mg/kg and slow waves at 9 mg/kg. Physostigmine decreased high voltage spindles and slow waves at 0.12 and 0.36 mg/kg. Based on the present results we propose that agonists possessing muscarinic M1 receptor activity are effective in decreasing high voltage spindles and scopolamine-induced slow wave activity, but agonists showing predominant muscarinic M2 receptor activity may be less effective in decreasing high voltage spindles and slow waves. Furthermore, tetrahydroaminoacridine decreased high voltage spindles at doses lower than those required to decrease scopolamine-induced slow waves. Physostigmine decreased high voltage spindles and slow waves over the same dose range. This result may indicate that non-cholinergic mechanisms are involved in the tetrahydroaminoacridine-induced decrease in high voltage spindles. PMID- 8405118 TI - A peptide muscarinic toxin from the Green Mamba venom shows agonist-like action in an inhibitory avoidance learning task. AB - A peptide, muscarinic toxin 2 (MTX2), isolated from Dendroaspis angusticeps venom was previously shown to displace the specific binding of [3H]pirenzepine, a muscarinic M1 receptor ligand, from rat brain synaptosomal membranes. We have tested MTX2 for muscarinic agonist or antagonist actions in an inhibitory avoidance task in rats. Infusion of the muscarinic receptor antagonist scopolamine into the hippocampus of rats immediately after the training period produced amnesia, whereas the muscarinic agonist oxotremorine increased retention. When MTX2 was injected into the hippocampus of rats after the inhibitory avoidance task, it caused memory facilitation, which could be suppressed by the concomitant infusion of scopolamine. Hence, in this test, MTX2 showed muscarinic receptor agonist-like actions, which are probably mediated by the M1 subtype of muscarinic acetylcholine receptors. PMID- 8405119 TI - The atypical antipsychotic drug amperozide enhances rat cortical and striatal dopamine efflux. AB - Previous behavioral, biochemical, and electrophysiological studies have indicated that amperozide, a putative atypical antipsychotic drug with potent 5-HT2 receptor antagonist potency, preferentially affects mesocorticolimbic, as compared to mesostriatal dopamine neurons. The present experiment utilized in vivo microdialysis to compare the effects of amperozide on dopamine efflux in medial prefrontal cortex versus caudate-putamen in the freely moving rat. The results demonstrated that amperozide produced a greater elevation of cortical dopamine. These results were similar to those observed with clozapine but not haloperidol. PMID- 8405120 TI - Cerebral cortex injury: effect of blockers of re-uptake of catecholamines. AB - In rats, surgical injury of the neocortex enhances the level of procholecystokinin-mRNA in the ipsilateral cortex. This increase in procholecystokinin gene expression was significantly reduced by the blockers of catecholamine re-uptake nomifensine (4 mg/kg), cocaine (5 mg/kg) and (+) oxaprotiline (1.5 mg/kg) given i.m. 30 min before the injury. The ganglionic blocking agent hexamethonium (3 mg/kg) prevented this effect of (+)-oxaprotiline and nomifensine. Also the alpha 1-adrenoceptor antagonists corynanthine (2 mg/kg) and prazosin (2 mg/kg) blocked this effect of (+/-)-oxaprotiline (1.5 mg/kg). It is concluded that catecholamines acting on alpha 1-adrenoceptors can reduce the increase in procholecystokinin-mRNA caused by cortex injury. The catecholamines may be released from the sympathetic nervous system. PMID- 8405121 TI - 4-Phenyltetrahydroisoquinoline, but not nomifensine or cocaine, inhibits methamphetamine-induced dopamine release. AB - The inhibitory effect of 4-phenyltetrahydroisoquinoline (4-PTIQ) on methamphetamine-induced dopamine release in the rat nucleus accumbens was investigated using a brain microdialysis method. Methamphetamine (10(-6) M) infusion through a microdialysis probe induced the release of dopamine. Although the uptake inhibitors, cocaine (3 x 10(-6) M) and nomifensine (10(-6) M), failed to block dopamine release, 4-PTIQ (10(-6 M) inhibited the dopamine-releasing effect of methamphetamine. 4-PTIQ did not affect the elevation of the extracellular dopamine level induced by high concentrations of nomifensine (10( 5) M) and cocaine (3 x 10(-5) M). 4-PTIQ was the weakest inhibitor of [3H]dopamine uptake by rat striatal synaptosomes. These results suggest that 4 PTIQ is a selective antagonist against the dopamine-releasing effect of methamphetamine in the nucleus accumbens. PMID- 8405122 TI - Ca2+(-)dependent and Ca2+(-)independent vasorelaxation induced by cardiotonic phosphodiesterase inhibitors. AB - Cardiotonic agents that belong to a group of phosphodiesterase inhibitors- vesnarinone, amrinone, enoximone, pimobendan, MS-857 and E-1020--inhibited the 35 mM KCl- and 10(-7) M norepinephrine-induced contractions of helical strips from rat thoracic aorta in a concentration-dependent manner. In the absence of extracellular Ca2+, 10(-7) M phorbol 12,13-dibutyrate caused a sustained contraction of the muscle strip without an increase in intracellular Ca2+ level ([Ca2+]i). The phorbol 12,13-dibutyrate-induced contractions in Ca2+(-)free buffer were also inhibited by the cardiotonic phosphodiesterase inhibitors. A cyclic GMP-inhibited cyclic nucleotide phosphodiesterase was partially purified from rat aorta and the activity of the phosphodiesterase was inhibited by the cardiotonic agents. The inhibitory effect of these agents on the KCl-, norepinephrine-and phorbol 12,13-dibutyrate-induced contractions showed good correlations to the concentrations of the agents producing 50% inhibition (IC50) of cyclic GMP-inhibited cyclic nucleotide phosphodiesterase. Vesnarinone inhibited the norepinephrine-induced contraction with a decrease in [Ca2+]i, but inhibited the phorbol 12,13-dibutyrate-induced contraction in Ca(2+)-free buffer without significant changes in [Ca2+]i. Dibutyryl cyclic AMP and NKH477 also inhibited the phorbol 12,13-dibutyrate-induced contraction in Ca(2+)-free buffer without significant changes in [Ca2+]i. The six cardiotonic phosphodiesterase inhibitors increased the cyclic AMP contents of the muscle strips. These results suggest that the inhibitory actions of these cardiotonic phosphodiesterase inhibitors on cyclic GMP-inhibited cyclic nucleotide phosphodiesterase may cause vasorelaxation through a decrease in [Ca2+]i and an inhibitory effect on a Ca(2+) independent contractile process (or a decrease in Ca(2+)-sensitivity of contractile elements). PMID- 8405123 TI - Involvement of delta-opioid receptors in physical dependence on butorphanol. AB - Butorphanol, a synthetic agonist/antagonist, has been shown to act on mu-, delta- and kappa-opioid receptors. However, the relative involvement of opioid receptor subtypes in mediating butorphanol dependence is not known. In the present study, naltrindole, a delta-selective non-peptide antagonist, was administered intracerebroventricularly (i.c.v.) to mask supraspinal delta-opioid receptors before and during the induction of butorphanol dependence. Treatment with naltrindole (0.1, 1, or 10 nmol/5 microliters per rat) significantly blocked naloxone-, a nonspecific antagonist, precipitated butorphanol withdrawal behaviors (escape behavior, teeth-chattering, wet shakes, forepaw tremors, ptosis, diarrhea, body weight loss, and hypothermia) at all doses tested, and decreased ejaculation at 0.1 nmol in butorphanol-infused rats. In contrast, naltrindole treatment had no effect on yawning, nor urination. These results indicate that central delta-opioid receptors are involved in mediating butorphanol dependence in rats. PMID- 8405124 TI - Human neuropeptide Y Y1 receptor antisense oligodeoxynucleotide specifically inhibits neuropeptide Y-evoked vasoconstriction. AB - This paper describes a new approach for the development of an inhibitor of the contractile responses of neuropeptide Y in human blood vessels by the use of an antisense oligodeoxynucleotide complementary to human neuropeptide Y Y1 receptor mRNA. One micromolar of an antisense 18-base oligodeoxynucleotide (hY1-AS), corresponding to the human Y1 receptor NH2-terminus, was incubated with segments of human subcutaneous arteries and veins for 48 h at 37 degrees C. Control vessels were incubated with the corresponding sense oligodeoxynucleotide (hY1-S) or a 3-base mismatched antisense oligodeoxynucleotide (hY1-MM) or no oligodeoxynucleotide. The contractile response to neuropeptide Y was markedly attenuated in both arteries and veins after treatment with hY1-AS, but was unaffected by hY1-S or hY1-MM. The pD2 values, i.e. the potency of neuropeptide Y, did not differ in hY1-AS treated vessels, suggesting a non-competitive receptor interaction as a result of down-regulation of Y1 receptors. Responses to noradrenaline or high K+ were unaffected by hY1-AS. This study may represent a new and highly specific approach to vascular pharmacology. PMID- 8405125 TI - Interleukin-8 is chemotactic for vascular smooth muscle cells. AB - The chemotactic activity of interleukin-8 for human aortic smooth muscle cells was investigated in Transwell cell culture chamber using a polycarbonate membrane with pores of 8 microns. Interleukin-8 stimulated the smooth muscle cell migration time and dose dependently in the range between 10(-11) and 10(-8) M. At 10 nM of interleukin-8, the number of migrated smooth muscle cells increased by 40-fold over the basal level, and this stimulation was almost completely abolished by anti-interleukin-8 antibody. Checkerboard analysis showed that the response of smooth muscle cell to interleukin-8 was chemotactic. These results suggest that interleukin-8 may play a role in the pathogenesis of arterial intimal thickening and atherosclerosis. PMID- 8405126 TI - Attenuation of hypercortisolemia in fawn-hooded rats by antidepressant drugs. AB - Long-term (21 days) treatment with imipramine, clomipramine (tricyclic antidepressants) and clorgyline (monoamine-oxidase type A inhibiting antidepressant) produced significant decreases in plasma corticosterone levels in fawn-hooded (FH) rats. In contrast, plasma adrenocorticotropic hormone (ACTH) levels were not altered by chronic imipramine or clorgyline treatment but were significantly higher in chronic clomipramine-treated FH rats. These findings demonstrate a differential effect of chronic antidepressant treatment on plasma ACTH and corticosterone concentrations in FH rats and, furthermore, support the results of earlier studies suggesting that the FH rat strain may represent a genetic model of depression. PMID- 8405127 TI - Verapamil suppresses d-amphetamine-induced place preference conditioning. AB - The effect of pretreatment with (+/-)-verapamil (5, 10 or 15 mg/kg, i.p.) on place preference induced with d-amphetamine (1 mg/kg, i.p. 40 min after verapamil) was studied in male rats. Place preference conditioning was performed using two-compartment shuttle boxes and 8 alternating stimulant/saline sessions. Verapamil dose-dependently suppressed amphetamine-induced place preference. No significant changes in place preference were observed following 8 alternating verapamil (no stimulant)/saline sessions, irrespective of whether verapamil injections were paired with the originally less or the originally more preferred compartment. It appears that verapamil effectively suppresses the reinforcing properties of d-amphetamine in the paradigm used. PMID- 8405128 TI - Role of 5-HT1-like receptors in the increase in external carotid blood flow induced by 5-hydroxytryptamine in the dog. AB - This study investigated the receptor involved in the 5-hydroxytryptamine (5-HT) induced increase in external carotid blood flow in pentobarbital-anaesthetized dogs. One-minute intracarotid (i.c.) infusions of 5-HT (0.3, 1, 3 and 10 micrograms) and 5-carboxamidotryptamine (5-CT; 0.01, 0.03, 0.1 and 0.3 micrograms) produced dose-dependent increases in external carotid blood flow without changes in mean arterial blood pressure or heart rate. After vagosympathectomy, the above vasodilator responses to 5-HT and 5-CT were abolished and remained so even after restoration of carotid vascular tone with noradrenaline. Furthermore, the 5-HT- and 5-CT-induced increases in external carotid blood flow were not modified by the 5-HT2 receptor antagonist, ritanserin (100 micrograms/kg i.v.), nor the 5-HT3 receptor antagonist, 1 alpha H,3 alpha, 5 alpha H-tropan-3yl-3,5-dichlorobenzoate (MDL 72222; 140 micrograms/kg i.v.), but were potently and dose dependently antagonized by the mixed 5-HT1-like and 5-HT2 receptor blocker, methiothepin (3, 10 and 30 micrograms/kg i.v.). Interestingly, the 5-HT1A and 5-HT1B receptor antagonist, cyanopindolol (100, 300 and 1000 micrograms/kg i.v.), blocked the effects of 5-HT, but the block was not elicited in a dose-dependent manner, with only the response induced by 0.3 microgram/min 5 CT being significantly antagonized by the highest dose of cyanopindolol; however, this blockade was not selective. Unlike 5-HT and 5-CT, 1 min i.c. infusions of either the 5-HT1C/5-HT2 receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl) aminopropane (DOI; 30-300 micrograms), or the 5-HT3 receptor agonist, 2-methyl-5 HT (10-300 micrograms), were devoid of effects on the canine external carotid blood flow. It is concluded that the 5-HT-induced increase in external carotid blood flow is mediated by 5-HT1-like receptors probably located on carotid sympathetic nerves. These receptors, however, do not seem to correspond to either the 5-HT1A, 5-HT1B or 5-HT1C receptor subtypes. PMID- 8405129 TI - D-myo-inositol 1,2,6-trisphosphate blocks neuropeptide Y-induced facilitation of noradrenaline-evoked vasoconstriction of the mesenteric bed. AB - Perfusion of the rat mesenteric bed with 0.1 or 10 nM neuropeptide Y potentiated the noradrenaline-induced increase in mesenteric pressure; the peptide did not modify basal perfusion pressure. While perfusion with 0.1 nM neuropeptide Y significantly increased the maximal noradrenaline-evoked vasoconstriction without modifying its EC50, 10 nM neuropeptide Y potentiated the maximal noradrenaline effect and significantly shifted its concentration-response curve to the left. Perfusion with 1-10 microM D-myo-inositol 1,2,6-trisphosphate (alpha-trinositol) reduced, in a concentration-dependent fashion, the neuropeptide Y-induced potentiation of the noradrenaline-evoked vasoconstriction without altering the potency or maximal response evoked by the catecholamine alone. Perfusion with 0.1 nM neuropeptide Y plus 1 microM alpha-trinositol completely abolished the neuropeptide Y-induced facilitation of the noradrenaline effect. alpha-Trinositol 1 microM in the presence of 10 nM neuropeptide Y caused a nonparallel rightward shift of the noradrenaline concentration-response curve as compared to that obtained in the presence of 10 nM neuropeptide Y alone. The alpha-trinositol blockade of the facilitatory action of neuropeptide Y was reversible. PMID- 8405130 TI - The role of the mu 2-opioid receptor in the antitussive effect of morphine in mu 1-opioid receptor-deficient CXBK mice. AB - The effect of morphine on the capsaicin-induced cough reflex was studied in mu 1 opioid receptor-deficient CXBK mice. There was no significant difference between the morphine-induced antitussive effect in CXBK mice and C57BL/6 mice, a progenitor strain. Furthermore, the antitussive effects of morphine in both the CXBK and C57BL/6 mice were antagonized by pretreatment with either naloxone or beta-funaltrexamine, a mu-opioid receptor antagonist, whereas pretreatment with naltrexonazine, a selective mu 1-opioid receptor antagonist, had no effect. Moreover, naltrindole, a selective delta-receptor antagonist, also had no significant effect on the antitussive effects of morphine in either CXBK or C57BL/6 mice. These results support our previous hypothesis that mu 2- rather than mu 1-opioid receptors are involved in morphine-induced antitussive effects. PMID- 8405131 TI - The hyoid bone position in adult individuals with open bite and normal occlusion. AB - The position of the hyoid bone was studied in 39 adult subjects with long face syndrome and an open bite of at least 2 mm (16 males and 23 females) and 43 adult subjects with normal Class I occlusion (25 males and 18 females). Several measurements were used to determine the horizontal, vertical, and axial orientation of the hyoid bone. Comparisons were based on paired t-test, which were performed separately for the male and the female groups. Most of the horizontal measurements showed no difference in the position of the hyoid bone in both male and female open bites when related to normals. As far as vertical measurements only the distance from hyoid bone to palatal plane was found significantly greater in the male open bite group due to the position of the palatal plane in the development of this dentoskeletal malocclusion. The hyoid axis formed significantly higher angles with the basion-nasion plane as well as with the palatal plane in the open bite groups, while there was no difference in the angle formed by hyoid axis and mandibular plane. The findings strongly suggest that hyoid bone moves in close conjunction with the pharynx, cervical spine, and mandibular plane in patients with entirely different skeletal patterns. PMID- 8405132 TI - Anterior open bite malocclusion: a follow-up study of orthodontic treatment effects. AB - Treatment response and stability of the anterior open bite malocclusion were evaluated in 20 open bite patients (17 females, 3 males), who were treated with the Edgewise appliance. The overbite, the number of teeth in occlusion, the functional occlusal contacts of the incisors and the cephalometric characteristics were studied before treatment (T1), at the end of retention (T2), and at least 1 year out of retention (T3). The apical root resorption of the upper incisors was analysed before and after treatment. The mean age at the follow-up control was 17 years 10 months. In 15 patients at least two occlusal contacts of the incisors were possible at the end of the retention period as well as at the follow-up control. The number of teeth in occlusion mesial to the second molars, expressed in percentage of the maximum possible, increased from 40 to 70 per cent during treatment with a positive tendency from T2 to T3. Apical root resorption of the upper incisors exceeded 10 per cent of the original root length in 4 out of 19 measurable cases. One of these patients had a history of trauma and the three others displayed atypical root form. Cases with an increased facial convexity, in which the uprighting of the incisors was possible, seemed to have a favourable treatment prognosis. PMID- 8405133 TI - The effect of routine steam autoclaving on orthodontic pliers. AB - Five commonly used types of orthodontic plier of similar design, but different manufacture were evaluated for wear and corrosion following a 6-month regime of routine use in a hospital department together with steam autoclaving in a centralized unit. Three different types of manufacture (metal finish) were evaluated: (1) a stainless steel plier; (2) a chrome plated plier; and (3) a cheaper chrome plated plier. Four identical kits for changing archwires were assembled for each of the three manufacture types. A sixth 'plier', always of chrome plated manufacture (group 2) was added to every kit and acted as an additional control. In all, 72 pliers were evaluated by three observers immediately before and after the trial. Visual analogue and rank scales were used to assess corrosion, damage, the efficiency of the plier, etc. Generally, all of the pliers stood up well to a combination of routine clinical use and steam autoclaving; however, the stainless steel pliers appeared to perform the best. It is likely that the most important factor, when setting up this type of sterilizing method for orthodontic instruments, is to establish a careful and meticulous routine for cleaning, lubricating, and steam autoclaving pliers. PMID- 8405134 TI - The effect of extraction of four second permanent molars on the incisor overbite. AB - Changes in incisor overbite were compared in 33 subjects who had four second molars extracted and 33 untreated subjects. There was an average increase in overbite of 0.7 mm in the extraction group, significantly different from an average decrease of -0.8 mm in the control group. The tendency to increase in overbite was greater in subjects whose presenting overbite was already reduced. The mechanism responsible for overbite reduction appeared to be a slight distal movement of the dentition, with retroclination of incisors and increase in inter incisal angle. PMID- 8405135 TI - Nasal airway, lip competence, and craniofacial morphology. AB - The aim of the present investigation was to demonstrate whether children selected by their open lip posture show differences in their nasal airway resistance and facial morphology compared to children with closed lip closure. Thirty-two children with poor lip competence were compared to a control group of 20 with secure mouth closure. Patients with poor lip competence showed a significantly higher ML-NSL angle and also ML-NL angle. Anterior rhinomanometry, as well as measurements of the pharyngeal space on lateral headfilms, displayed no significant differences between the two groups. The same material was also grouped cephalometrically into skeletal open and skeletal deep bite configurations. The mean value of the nasal airway passage was lower in the skeletal open group, but not at a significant level. PMID- 8405136 TI - A comparative assessment of cephalometric errors. AB - Measuring a cephalometric radiograph is an imprecise task, and the errors associated with this should be quantified and understood. The potential sources of error are outlined and the literature relating to the assessment of measurement errors is reviewed. Three techniques are examined: the error attributable to a single recording; its associated confidence limits; and the coefficient of reliability were the most commonly applied. Data from two error studies were pooled and used to compare the accuracy of 12 skeletal, dental, and soft tissue measurements, using various assessments of each of the three techniques. Within each group, the use of different mathematical criteria led to variation in the results; one method was totally unsuitable. As with all statistical procedures, a knowledge of the technique facilitates interpretation of the results. Some suggestions as to the most satisfactory approach to the estimation of measurement error in cephalometry are given. PMID- 8405137 TI - A cephalometric comparison of medieval skulls with a modern population. AB - Thirty-one medieval skulls, mostly dating from the Black Death in 1348, were compared with 32 modern, untreated adults. The medieval population was unusual because the skulls could be so closely dated. Lateral skull radiographs were used for the comparison, based on the method of Seddon (1984). A method error study indicated that the measurements could be treated with reasonable confidence, although possible difficulties with the material are discussed. The findings suggest that the modern group have the longer face and longer palate. The medieval group were found to have more retroclined upper incisors and more proclined lower incisors as well as reduced overbites. The results also suggest changes in mandibular shape over time together with cranial base changes: the modern mandible has a more obtuse gonial angle and is generally longer and thinner. It was not possible to reach a firm conclusion regarding chin prominence. The cranial base saddle angle has become more acute since the medieval period, and the length S-N is greater in the modern group. Depending on interpretation, the skull material could be said to have a more prominent maxilla. Possible reasons for the differences are discussed. PMID- 8405138 TI - Protective influence of spayed and androgen-treated females on oestrous cycle in food-restricted mice. AB - Diet restriction induced disruption of oestrous cycle in regularly cycling female mice. The irregularities in the oestrous cycle in females were directly related to the degree of diet restriction. The presence of spayed and androgen-treated females prevented these irregularities and helped to maintain regular oestrous cycles in underfed females. The ability of androgen-treated females to prevent the irregularities in the oestrous cycle in underfed females was inversely related to the degree of diet restriction. The results suggest that the production of the male-originating pheromone that helps to prevent the underfeeding-induced disruption of oestrous cycles in females is androgen dependent. PMID- 8405139 TI - Quality of life in patients with Addison's disease: effects of different cortisol replacement modes. AB - This study compares the impact of different modes of cortisol replacement therapy on the health perception and general well-being in patients with primary adrenocortical failure. 14 adults (8 female, 6 male) with Addison's disease on chronic cortisol replacement participated in the study. In a randomized double blind cross-over design, all patients were treated with 3 modes of cortisol replacement for one week each (mode I: 20 mg hydrocortisone (HC) at 0700 h and 10 mg HC at 1900 h; mode II: 30 mg HC at 0700 h and placebo at 1900 h; mode III: placebo at 0700 h and 30 mg HC at 1900 h). Following the third week, the replacement modes were repeated in a different random order. For quality-of-life assessment the patients completed three different questionnaires (Addison questionnaire, Basler Befindlichkeits-Skala, Beschwerde-Liste) and were interviewed about their general contentment at the last day of each treatment week. General well-being in terms of subjective contentment was best established during mode I (in 64% of patients) and less often stated with mode II (in 29%) and III (in 14%) (p < 0.05 mode I vs III). With the twice-daily replacement (mode I), sum scores of all questionnaires were changed towards improvement compared to both once-daily regimens (p < 0.05 vs mode II and III), but did not reach normal values of healthy subjects. Differences between mode II and III were insignificant. We conclude that quality of life in Addison patients is mainly influenced by the mode of cortisol replacement therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405140 TI - Salt-losing form of 21-hydroxylase deficiency accompanied by hypopituitarism in a boy. PMID- 8405141 TI - Molecular genetic analysis of NIDDM. Status and prospects. PMID- 8405142 TI - Functional properties of isolated stroma and epithelium from rat ventral prostate during androgen deprivation and estrogen treatment. AB - To identify the functional activities of prostatic stroma under different hormonal conditions, isolated stroma and epithelium from rat ventral prostate (RVP, intact or one week castrated or estrogen-treated), were studied in metabolic labeling experiments. Using a semiquantitative stereological procedure, the relative proportion of the epithelial and stromal compartment was determined in situ. In addition, the distribution of the androgen receptor was visualized by in situ hybridization and by immunocytochemistry. In castrated animals protein biosynthesis of the stroma and epithelium exceeded the control value by a factor 7 and 5, respectively. In estrogen-treated animals protein biosynthesis was reduced, reaching only between one tenth and one fifth of the control value. The amount of stroma obtained from these animals was very low. These results were confirmed by stereological findings and indicate a differential regulation of prostatic stroma and epithelium after estrogen challenge and androgen deprivation. Estrogen receptor was induced in epithelium and stroma in estrogenized animals whereas the androgen receptor was reduced in experimental specimens. During estrogenization the biosynthetic activity of both stroma and epithelium is depressed, while estrogen responsivity of the epithelium in terms of estrogen receptor expression is increased. Androgen withdrawal results in active transformation of the gland through increased stromal biosynthetic activity and epithelial regression. PMID- 8405143 TI - Stromal and epithelial cells from rat ventral prostate during androgen deprivation and estrogen treatment--regulation of transcription. AB - To identify the functional capacities of prostatic tissue, the expression of steroid hormone receptors, growth factors, oncogenes and particular enzymes was studied at the RNA level in isolated stromal and epithelial cells of rat ventral prostate (RVP) under different hormonal conditions (androgen deprivation, estrogen treatment). Slot blot and Northern blot analyses of isolated RNA resulted in characteristic changes: In the control prostate, androgen receptor (AR) mRNA was high in epithelium of intact prostate, but low in stroma. Its level was increased after castration in the epithelium during the initial 24 hours, whereas an only slight increase occurred in stroma after one week castration. The AR signal was not altered by estradiol treatment in epithelium and stroma. Conversely, the estrogen receptor (ER) mRNA, predominant in stroma and very low in epithelium, decreased after castration in stroma and epithelium within 24 hours and was absent one week later. After estrogen treatment the ER signal increased considerably in stroma. mRNA of both basic fibroblast growth factor (bFGF) and transforming growth factor beta (TGF-beta) were exclusively found in stroma. Both were low in controls and responded in a different way to castration and estrogen treatment within 24 hours. bFGF was increased in estrogen-treatment animals, while TGF-beta was induced by castration. Shortly after castration (2 hours) v-fos expression increased and reached a maximum after 6 hours, but was no more detectable after 12 hours in epithelium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405144 TI - Effects of selenium and iodine deficiency on type I, type II and type III iodothyronine deiodinases and circulating thyroid hormones in the rat. AB - The effects of nutritional selenium (Se) deficiency over a period of three generations and of a combined selenium and iodine deficiency on hepatic and cerebrocortical iodothyronine deiodinases and on circulating thyroid hormone levels were examined in the rat. Se deficiency strongly decreased hepatic type I iodothyronine 5'- and 5-deiodinase to 6-13% of that in controls. Iodine depletion had only a marginal decreasing effect on the type I activity. Cerebrocortical type II 5'-deiodinase was decreased in Se-deficient, iodine-replete rats. Its 5-6 fold elevation in iodine-deficient rats was not reversed by additional selenium deficiency. Cortex type III 5-deiodinase was modestly decreased in all groups with insufficient trace element supply. Long-term Se deficiency has only limited effects on serum T4 and T3 levels. Two months of iodine deficiency decreased serum T4 to less than 10% of that in controls, but did not significantly affect serum T3 levels. The strong decrease of hepatic outer- and inner-ring deiodination of T4 in Se deficiency obviously reflects the reduced tissue concentration of the type I deiodinase which was recently identified as a selenoenzyme. The maintenance of increased cerebrocortical type II deiodinase in iodine-depleted animals irrespective of adequate or deficient selenium supply suggests that the type II isoenzyme does not contain selenium in its catalytic site. Further studies are necessary to clarify whether the weak, but repeatedly confirmed decrease of cortex type III deiodinase is the direct effect of Se deficiency or the indirect consequence of the multilevel change in thyroid hormone metabolism.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405145 TI - The stimulation of rat Leydig cell steroidogenesis by human ovarian steroidogenesis-inducing protein (SIP) may not require endogenous cAMP. AB - Recently a protein from ovarian follicular fluid was isolated which stimulates steroid production in different cells (Khan et al., 1990). The present study was performed to further characterize the short term effects of this steroidogensis inducing protein (SIP) on steroid production in isolated rat Leydig cells and to compare the effects with LH/hCG and LHRH. SIP stimulated testosterone production in a dose-dependent manner. In Leydig cells isolated from adult rats the degree of stimulation was much higher than that obtained with hCG, dibutyryl cAMP (db cAMP) or LHRH. Moreover, the stimulated steroid production in the presence of hCG or db cAMP was further enhanced by SIP. The time courses of hCG and SIP action on testosterone production were comparable and maximal stimulation of steroid production was obtained within one hour. In contrast to hCG, SIP did not stimulate cAMP production. An antagonist of LHRH action was unable to block the effects of SIP on Leydig cells indicating that SIP does not act via LHRH receptors. Neither SIP nor LH could further stimulate the steroid production in the presence of 22-R-OH-cholesterol, illustrating that both stimulators control the availability of cholesterol as substrate. An inhibitor of mitochondrial cholesterol side chain cleavage (CSCC), aminoglutethemide, completely blocked the stimulatory effects of SIP and LH/hCG. Thus the effects of SIP on steroid production are not the result of conversion of contaminating steroids in the SIP preparation. SIP and LH/hCG actions were also blocked when the cells were incubated in the presence of cycloheximide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405146 TI - Verotoxin producing Escherichia coli O 157 infections associated with the consumption of yoghurt. AB - Sixteen cases of verotoxin producing Escherichia coli (VTEC) O 157:H7 Phage Type 49 infection were identified in the North West of England from 1 September to 1 November 1991, eight of whom lived in or around the same large town. Eleven of the cases were aged 10 years or less, and five of the affected children developed haemolytic uraemic syndrome. A case control study demonstrated a strong association between VTEC O 157:H7 PT 49 infection and the consumption of a locally produced live yoghurt. This is the first time that an outbreak of VTEC O 157 infection has been linked to the consumption of yoghurt and this vehicle of infection should be considered when investigating such outbreaks in future. PMID- 8405147 TI - A comparison of multiple drug resistance in salmonellas from humans and food animals in England and Wales, 1981 and 1990. AB - For Salmonella typhimurium from humans in England and Wales, the incidence of multiple resistance more than doubled over the 8-year period 1981-8 and, over the next 2 years, increased by a further 7%. From 1981 to 1988 both resistance and multiple resistance also increased significantly in S. virchow and although multiple resistance did not increase over the next 2 years, the overall incidence of resistance has continued to rise. In 1990 the majority of S. typhimurium from cattle were multiply-resistant and the occurrence of such resistance has quadrupled since 1981. Multiple resistance has also increased in S. typhimurium from pigs and, to a lesser extent, from poultry. In contrast, multiple resistance has remained uncommon in the poultry-associated serotype S. enteritidis. For S. virchow, multiple resistance was common in a phage type frequently associated with poultry meat imported from France. The continuing use of a range of different antimicrobials in calf husbandry has been an important factor in promoting the emergence of multiply-resistant strains of S. typhimurium in cattle. In contrast, multiple resistance has remained rare in those serotypes associated with poultry, where the use of such antimicrobials has been less intensive. It is hoped that recent recommendations discouraging, in veterinary medicine, the prophylactic use of antibiotics with cross resistance to those used in human medicine will result in a reduction in the occurrence of multiresistant strains in food animals and subsequently in humans. PMID- 8405148 TI - Variations in biochemical phenotypes and phage types of Salmonella enteritidis in Germany 1980-92. AB - The Phene Plate system for typing Salmonella serotypes (PhP-S) is a simple automated typing method based on biochemical fingerprinting. It gives a quantitative value of the metabolism of various substrates by measuring the speed and intensity of each reaction. The 'biochemical fingerprint' of each isolate is used to calculate similarities among the tested strains with a personal computer program. We used this system to examine a collection of 86 strains of Salmonella enteritidis isolated from human sporadic cases in Germany between 1980 and 1992. Twenty-three biochemical phenotypes (BPTs) consisting of 9 common (C) and 14 single (S) BPTs were identified. BPTs C2 and C4 containing 20 and 36 strains respectively accounted for 65% of the isolates. Strains of BPT C2 were found over a wide period of time whereas strains of BPT C4 were isolated during the period between 1988 and 1992. With phage typing, 11 discrete phage types (PTs) and 18 strains designated as non-specific type (NST) were identified. PTs 4 and 8 with 39 and 17 strains respectively were the dominant PTs. Strains of PT 8 were isolated over a wide period of time whereas all (except one) strains of PT 4 were isolated between 1988 and 1992. Combination of biochemical fingerprinting and phage typing divided the strains into 25 phenotypes (BPT:PTs). Whilst phenotype C2:8 was found over a number of different years, phenotype C4:4 was isolated only between 1988 and 1992. These findings indicate the presence of one persistent and one recently emerged phenotype among S. enteritidis strains in Germany.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405149 TI - Egg age and the growth of Salmonella enteritidis PT4 in egg contents. AB - The growth of Salmonella enteritidis PT4 in albumen around an intact yolk was governed by the age of the egg on inoculation. In the majority of eggs, held at 20 degrees C, the bacterium was unable to grow rapidly until eggs had been stored for approximately 3 weeks. The multiplication of S. enteritidis in stored eggs appeared to be associated with alterations to the yolk membrane which allowed the bacterium to either invade the yolk or obtain nutrients from it. The rate at which egg contents change to permit the growth of S. enteritidis would appear to be temperature related and took place more rapidly when eggs were stored under conditions where temperatures fluctuated and, on occasions, reached 30 degrees C. PMID- 8405150 TI - The prevalence of antimicrobial resistance in human faecal flora in South Africa. AB - Between January and March 1992, 361 faecal specimens were collected from the healthy black population in the Transvaal Province of South Africa. Each specimen was examined for the prevalence of antimicrobial resistance in commensal bacteria. Volunteers, from both rural and urban dwellings, were divided into four age groups. The overall carriage rate of resistance varied from 88.6% for ampicillin, 74.2% for trimethoprim, 52.6% for chloramphenicol, 10.2% for nalidixic acid to 7.5% for gentamicin. The carriage of resistance found to each individual antimicrobial agent was slightly higher in the rural population rather than the urban population but there was no correlation between the prevalence of antimicrobial resistance and the age group. PMID- 8405151 TI - Replicon typing characterization of plasmids encoding resistance to gentamicin and apramycin in Escherichia coli and Salmonella typhimurium isolated from human and animal sources in Belgium. AB - Escherichia coli and salmonella strains with plasmids conferring resistance to gentamicin and apramycin have been isolated with increasing frequency both from cattle and hospital patients in Belgium. The apramycin-gentamicin resistance plasmids were characterized in recipient strains by their profiles and molecular weights using agarose gel electrophoresis, by their antimicrobial resistance patterns and by replicon typing using a series of DNA probes specific for the genes controlling their systems of replication. Overall, most of the plasmids differed in their DNA electrophoretic patterns. Seventeen different antimicrobial resistance profiles were observed, and there were six different types of replicons. However, two replication genes predominated and had a preferential distribution in different bacterial species. The rep FIC.a plus rep Q multireplicon was found mainly in plasmids recovered from gentamicin- and apramycin-resistant E. coli while replicon of the type rep FIC.b largely prevailed in S. typhimurium. Identical replication genes were found in most animal and human strains, hence suggesting a high homology between apramycin gentamicin plasmids in these communities. Finally, our results indicate that the rapid spread of apramycin-gentamicin-resistance in several species of Enterobacteriaceae isolated from animals and from humans in Belgium is not due to a single plasmid, but rather that the gene encoding AAC(3)-IV is carried by various replicons. PMID- 8405152 TI - Epidemiological investigation of risk factors for campylobacter colonization in Norwegian broiler flocks. AB - An epidemiological investigation was conducted to identify risk factors related to hygiene and husbandry practices which determine the introduction of Campylobacter spp. into broiler chicken flocks. All 176 broiler farms in an area in southeastern Norway participated in the study. Each farm was represented by one flock selected at random during a one-year period. The flocks were examined for campylobacter colonization at slaughter, and the flock managers were subsequently interviewed about hygiene and husbandry practices. Campylobacter spp. were recovered from 32 (18%) of the flocks. The proportion of colonized flocks varied geographically and seasonally with a peak in the autumn. The following variables were found to be independently associated with an increased risk of campylobacter colonization using logistic regression analysis: (i) feeding the broilers undisinfected water (odds ratio (OR) = 3.42, P = 0.045), (ii) tending other poultry prior to entering the broiler house (OR = 6.43, P = 0.007), (iii) tending pigs before entering the house (OR = 4.86, P = 0.037), (iv) geographic region (Hedmark versus Ostfold county) (OR = 2.91, P = 0.023, (v) season (autumn versus other seasons) (OR = 3.43; P = 0.008). Presence of rats on the farm was associated with an increased risk, but this factor did not reach statistical significance (OR = 3.96, P = 0.083). Preventive measures should include disinfection of drinking water and strict hygienic routines when the farm workers enter the rearing room. The results indicate that disinfection of drinking water is the preventive measure most likely to have the greatest impact on the prevalence of campylobacter among broiler chicken flocks in the study area (population attributable fraction = 0.53). PMID- 8405153 TI - Characterization of human faecal flora by means of an improved fluoro morphometrical method. AB - The use of polyclonal antibodies in differentiating between samples of faecal microflora derived from ten healthy volunteers was assessed. The distribution of FITC-labelled antibodies (of the IgA, IgG and IgM isotype) over 144 morphologically distinct subsets of faecal bacteria ('morphotypes') was quantified by means of digital morphometry and quantitative immunofluorescence. Furthermore, a new dataprocessing algorithm was developed which makes objective quantitation of the antibody distributions over the faecal morphotypes possible. The results of this study imply that the antibody binding capacity of faecal morphotypes is a unique characteristic of faecal microflora. PMID- 8405154 TI - Healthy individuals possess circulating antibodies against their indigenous faecal microflora as well as against allogenous faecal microflora: an immunomorphometrical study. AB - Healthy persons were shown to possess circulating antibodies of both IgA, IgG and IgM isotype directed against the bacteria of their faecal microflora, assessed by immunomorphometry. After removal, by absorption, of the fraction of antibodies directed against the autochthonous faecal bacteria or cross-reacting with allogenous faecal bacteria, there were still antibodies left directed against allogenous faecal bacteria of both the IgA, IgG and IgM isotype. However, relatively more antibodies of the IgA isotype appeared to be directed against allogenous bacteria than against indigenous faecal bacteria. Persons who reacted with specific antibodies to many bacteria of their own flora also tended to react specifically to bacteria in the allogenous microflora of the other volunteers. The patterns of antibodies directed to faecal bacteria of different morphologies (morphotypes) were unique for each individual. PMID- 8405155 TI - A model for autumn outbreaks of Legionnaires' disease associated with cooling towers, linked to system operation and size. AB - Cooling towers have been demonstrated to be amplifiers and disseminators of legionella, the causative organism of Legionnaires' disease. Community outbreaks associated with cooling towers have been reported with several common factors. Small towers (< 300 kW) have predominantly been implicated in outbreaks. Cooling tower-associated outbreaks are most frequent in autumn, and frequently implicated systems have been operated after a period of shutdown. This paper reports field study data relating system operation to legionella colonization of systems. Operating systems have been shown to be more frequently colonized by legionella than shutdown systems. In some cases operation of systems after periods of shutdown raised legionella concentrations from below detection limits to between 50 and 950 c.f.u./ml within 10 min. These data and previously reported data relating to biofilm and sediment colonization of the systems, and community outbreaks of Legionnaires' disease, have been used to develop a model explaining the seasonal nature of outbreaks associated with irregularly operated, small cooling tower systems. PMID- 8405156 TI - The epidemiology and surveillance of visceral leishmaniasis in the Campania region of Italy. The value of zymodeme typing. AB - Although human visceral leishmaniasis (VL) is a notifiable disease in Italy, there is evidence that the actual number of cases is far higher than that notified. A programme for active surveillance of VL in the 14 Italian endemic regions was launched by the Istituto Superiore di Sanita. We report data collected during a 3-year period of active surveillance in Campania, a south Tyrrhenian region covering 4.5% of the Italian territory. Out of 120 clinically suspected cases referred to medical and diagnostic references centres, there were 52 confirmed VL cases (17.3/year), i.e. 10-fold more than previously notified. Most of the infection sites were in rural areas or peripheral districts of towns in hilly parts of Naples province. An epidemic cluster of 10 cases was identified in a microfocus of Caserta province. The biochemical analysis of 23 Leishmania stocks showed a zymodeme composition indicating Campania as an old and well established focus of VL. The data obtained emphasize that the present notification system for VL in Italy is inadequate. PMID- 8405157 TI - Mycoplasma pulmonis infection of the murine oropharynx protects against subsequent vaginal colonization. AB - Intranasal inoculation of 12 young adult mice (strain TO) with Mycoplasma pulmonis protected all of them against vaginal colonization when they were challenged intravaginally 60 days later with the same mycoplasmal strain. In contrast, all 15 mice without a respiratory infection became colonized vaginally (geometric mean titre [GMT] 4.6 x 10(6) colour-changing units [c.c.u.]) when challenged in the same way. The GMT of serum antibody, measured by a microimmunofluorescence technique, prior to challenge was 200 and 8 for the oropharyngeally infected and unexposed mice, respectively. The GMT of antibody in vaginal washings from the two groups was 6 and 3, respectively. All four nude BALB/c mice were susceptible to vaginal colonization (GMT 5.6 x 10(6) c.c.u.) after oropharyngeal infection (GMT 5.1 x 10(4) c.c.u.) resulting from intranasal inoculation, as were all six nude mice (vaginal GMT 1.4 x 10(7) c.c.u.) that had not been inoculated intranasally. In contrast, all ten of their immunocompetent counterparts were resistant to vaginal colonization after oropharyngeal infection (GMT 1.3 x 10(3) c.c.u.), whereas all nine such mice that had not been infected oropharyngeally were susceptible to vaginal colonization (GMT 7.6 x 10(6) c.c.u.). These results show the important role that a respiratory infection has in protecting the vagina against colonization and that protection is dependent on a functioning T-lymphocyte system. PMID- 8405158 TI - The influence of local antichlamydial antibody on the acquisition and persistence of human ocular chlamydial infection: IgG antibodies are not protective. AB - In order to study the effect of antichlamydial antibodies in ocular secretions on resistance to ocular chlamydial infection and clearance of this infection, we have performed linked longitudinal studies in a Gambian village in which trachoma is endemic. We have measured IgG and IgA antibody levels to a local serotype B isolate of Chlamydia trachomatis by amplified enzyme immunoassay, and chlamydial antigen levels in conjunctival swabs using a commercially available immunoassay which detects chlamydial glycolipid. Having previously demonstrated that sharing a bedroom with a case of active trachoma is a risk factor for acquisition of the disease, we have analyzed the effect of IgG and IgA antibody on the acquisition and persistence of clinical trachoma after controlling for age, sex, exposure to infection and for the presence of chlamydial antigen using a Poisson regression model. We have found that the presence of antichlamydial IgG in ocular secretions of disease-free subjects is associated with an increased incidence of trachoma. IgA antibody shows an opposite trend, but this is not statistically significant. One possible explanation of these findings is that antichlamydial IgG antibodies enhance the infectivity of C. trachomatis for the human eye; this could have major implications for the development of a chlamydial vaccine. PMID- 8405159 TI - Bacteriophage as models for virus removal from Pacific oysters (Crassostrea gigas) during re-laying. AB - A study was undertaken to examine the feasibility of using naturally-occurring bacteriophages to assess the impact of re-laying on levels of viral contamination in Crassostrea gigas, the Pacific oyster. Two phages were chosen. One, male specific (F+), was enumerated using Salmonella typhimurium. The other, a somatic phage, was detected using an, as yet, uncharacterized Escherichia coli. Investigations, using a variety of re-laying sites, demonstrated that numbers of F+ phage in oyster tissue declined more rapidly than those of somatic phage. For example, in oysters placed in commercially-used sea water ponds, F+ phage reached undetectable levels within 2-3 weeks, whereas somatic phage could still be detected 5 weeks after re-laying. The studies suggest that F+ phage may not be a suitable indicator for virus removal and that somatic phage may be better suited to this role. PMID- 8405160 TI - Impact of respiratory virus infection in patients with chronic chest disease. AB - This study investigated the morbidity associated with respiratory virus infections in patients with well-documented chest disease, and the risk of transmission between close contacts. Patients informed the study team if they were exposed to a family member or colleague with a cold. Patients and symptomatic index cases recorded respiratory symptoms during the study period. Acute nasopharyngeal swabs and paired sera were obtained for viral diagnosis. Twenty-five (43%) of 58 recorded exposures resulted in a symptomatic illness and 16 (28%) patients developed lower respiratory tract symptoms. Sixteen (64%) of the 25 symptomatic patients contacted their general practitioner, 14 (56%) received antibiotics and 4 (16%) were hospitalized. Mean duration of illness was 10.6 days in symptomatic patients and 5.7 days in their corresponding index cases (P < 0.005). Mean symptom scores were 100.6 in symptomatic patients and 62.2 in index cases (P < 0.01). Respiratory viruses were identified in 19 (33%) episodes. Rhinovirus, coronavirus and respiratory syncytial virus infections were all associated with lower respiratory tract exacerbations. Respiratory tract symptoms following exposure to a cold were comparatively severe in these patients with chronic chest disease. This group of patients might gain particular benefit from the introduction of effective vaccines or antiviral therapy. PMID- 8405161 TI - Influenza immunization; vaccine offer, request and uptake in high-risk patients during the 1991/2 season. AB - Current levels of influenza vaccine uptake in patients considered to be at high risk have been determined by means of a questionnaire survey. During March-April 1992, information was sought from 624 patients in Leicestershire, UK with either chronic cardiovascular or respiratory disease, or diabetes; questions related to current health status and the request, offer and receipt of influenza vaccine in the current and three previous seasons. Ninety-eight percent of all offers of immunization were made in the primary care setting, and vaccine was well tolerated as judged by the fact that 86% of vaccinees between 1988/9-1990/1 returned for immunization in the following year. However in the 1991/2 season the overall level of vaccine uptake was only about 41% which is at variance with the stated policies and practices of general practitioners. Opportunities were missed, in both hospitals and general practices, to publicise and offer immunization to individuals with vaccine indications. Future attempts to improve vaccine uptake should focus on increasing the role of hospital staff in influenza prevention, in addition to promoting better vaccine delivery through primary care. PMID- 8405162 TI - Laboratory surveillance of viral meningitis by examination of cerebrospinal fluid in Cape Town, 1981-9. AB - Nine years accumulated laboratory data derived from the culture of the cerebrospinal fluid of 11,360 aseptic meningitis cases were retrospectively reviewed to establish the epidemiology of viral meningitis in Cape Town. Virus was isolated from 3406 of the cases (91% enteroviruses and 9% mumps). Five major summer viral meningitis episodes were documented: two of echovirus 4 (706 and 445 cases), echovirus 9 (223), coxsackie A9 (104) and one of unidentified enterovirus (324 cases--probably echo 9). Although coxsackie B was endemic, clusters of one or other type were dominant at any one time. Mumps was endemic. Sixty-two percent of all viral cases were < 5 years old. The median ages of 4 and 5 years in echoviruses 9 and 4 (the epidemic strains) contrasted with that of 1 year in coxsackie B (with many cases < 3 months old). Mumps peaked at 3-4 years of age. Males dominated overall, particularly in mumps. PMID- 8405163 TI - Weather factors in the prediction of western equine encephalitis epidemics in Manitoba. AB - Cases of western equine encephalitis in horses in 1987 in western USA and Manitoba, Canada were examined by backward trajectory analysis of winds. Culex tarsalis mosquitoes infected with western equine encephalitis virus could have been carried on southerly winds from Texas and Oklahoma to northern USA and from there to Manitoba. The presence of the Polar front over North Dakota and Minnesota at the end of July would have led to the landing of Cx. tarsalis in Montana and Wisconsin and prevented further carriage into Manitoba. Temperatures in southern Texas during the winter months (average daily maximum temperatures 19.7 degrees C and higher) would have permitted continuous transmission of western equine encephalitis virus by Cx. tarsalis in this area. Weather factors involved in outbreaks from 1975-88 were analysed to see if epidemics in Manitoba (23 or more cases in horses) could be predicted. The conditions for epidemics could be defined as follows: (a) the number of cases in horses in USA was 98 or more, (b) winds were southerly with speeds 45 kmh-1 or higher, and (c) counts of Cx. tarsalis females/light trap per day were 3.2 or higher. There were 3 or fewer cases in Manitoba, when the number of cases in USA was 27 or less, even when Cx. tarsalis counts were higher than 3.2. With Cx. tarsalis counts below 3 and/or unsuitable winds, or the Polar front further south, the number of cases in Manitoba was between 0 and 17, even when the number of cases in USA was from 38 172. Without information on the extent of infection further south, the weather variables would probably be more useful in excluding the possibility of an epidemic in Manitoba than in predicting one. PMID- 8405164 TI - Distinct yearly change of serotype distribution of human rotavirus in Thailand as determined by ELISA and PCR. AB - A total of 241 group A rotavirus-positive stool samples collected from diarrhoeic patients in Thailand between July 1988 and June 1991 were characterized for their serotypes by enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies and by a polymerase chain reaction (PCR). In July 1988-June 1989, serotype 1 was the most prevalent (63.4%), followed by serotype 4 (11.0%) and serotype 2 (8.5%). In July 1989-June 1990, 59.8% were serotype 1, 24.3% were serotype 2, and 6.1% were serotype 3. In contrast, in July 1990-June 1991, serotype 3 was detected in the highest frequency (40.5%), 29.9% were serotype 1, and 27.3% were serotype 2. Thus, a distinct yearly change of serotype distribution of rotavirus in Thailand was observed in the three consecutive years. In particular, it was of note that the prevalence of serotype 3 greatly increased, in contrast to the previous studies in which almost no serotype 3 rotaviruses were detected in the years 1983-8 in Thailand. PMID- 8405165 TI - Infrared study of the structure and composition of rabbit lens membranes: a comparative analysis of the lipids of the nucleus, cortex and epithelium. AB - Infrared spectra of the phospholipids extracted from the nuclear, cortical and epithelial regions of the rabbit lens membrane bilayers are examined for the first time. Major structural, conformational and compositional differences in the lens membrane are correlated at physiological temperature. Vibrational data distinguish two classes of the phospholipids present in the rabbit lens membranes. Sphingolipids with a sphingosine backbone are largely concentrated in the nucleus whereas phospholipids with a glycerol backbone (glycero-lipids) such as phosphatidylcholine and phosphatidylethanolamine are abundant in the cortical region of the lens. Nuclear lipids are more saturated by a factor of four over the lipids of the cortical region. Nuclear lipids also exhibit increased headgroup and interface hydration and stronger hydrogen bonding over cortical lipids which provide them additional structural stability needed for the clarity of the lens. The lipid composition of the epithelial membranes is structurally similar to that of the cortical region but the hydrocarbon chains are more saturated. Epithelial membranes are largely shielded from bilayer water indicating hydrophilic lipid-water interactions are not important for the stability of these membranes. These membranes exhibit much stronger lipid-lipid interactions, vital for many physical properties like membrane fluidity and permeability. The fiber cell membranes are stabilized by hydrogen bonding of the carbonyls of the interface region and by headgroup-specific lipid-water interactions, while epithelial membranes are stabilized primarily through hydrophobic lipid-lipid interactions. Data also exhibit two phase transitions around 16 and 35 degrees C corresponding to the melting of the hydrocarbon chains from two pools of the phospholipids i.e. glycero- and sphingophospholipids, respectively. The first transition is caused by disruption of the carbonyl hydrogen bonding and disordering of the acyl chains of the glycero-lipids, while second transition is driven mainly from the hydrogen bonding effects of the carbonyls of the sphingolipids followed by acyl/sphingosine chain disordering. Upon transition to the liquid-crystalline phase, a sizable amount of the fiber cell membranes are disordered (approximately 33%) due to increased conformational motion of the acyl chains resulting from a loss of extended CH2 trans segments. These chains of the fully hydrated lipids pack in an hexagonal or triclinic unit cell in rabbit lens membranes. PMID- 8405166 TI - Corneal PAF acetylhydrolase increases in anterior segment ischemia in rabbits. AB - The involvement of platelet-activating factor (PAF) in the development of anterior segment necrosis after occlusion of the bilateral long posterior ciliary arteries was investigated in rabbits. With the progression of anterior segment necrosis, which was characterized by corneal edema and neovascularization, the contents of protein and total phospholipids increased in the cornea and aqueous humor, but not in the iris. PAF acetylhydrolase activity was significantly increased on the first postoperative day in the peripheral cornea and on the second day in the central cornea prior to the development of marked corneal edema and neovascularization, but it did not increase in the iris. The accumulation of newly synthesized PAF in the cornea reached a plateau level on the third postoperative day, which coincided with the progression of the corneal lesion. In the aqueous humor, PAF acetylhydrolase activity was increased by the induced ischemia, but no PAF was detected. The increased PAF acetylhydrolase activity may reflect adaptation to the participation of PAF in the progression of anterior segment necrosis. PMID- 8405167 TI - Light-activation of teleost rod photoreceptor elongation. AB - Rod photoreceptors in the retinas of teleost fish undergo changes in cell length in response to changing ambient light intensities. In the dark rods shorten and in the light rods elongate. These movements are mediated by actin-dependent processes which occur in the ellipsoid and myoid of the inner segment. As an approach to examining the underlying intracellular signaling pathways that link light absorption to actin-dependent motility in the inner segment, we have investigated the quantitative aspects of the light stimulus required to activate elongation in isolated rod inner/outer segments (RIS-ROS) of the green sunfish (Lepomis cyanellus). The intensity thresholds and strength-duration characteristics of the light stimulus required to activate teleost rod elongation were found to differ from those reported to activate vertebrate rod membrane hyperpolarization. In response to brief pulses of light, RIS-ROS elongated in a graded manner, both as a function of increasing light pulse intensity and light pulse duration. Half maximal activation of light-induced RIS-ROS elongation was produced by a stimulus of roughly 6 x 10(15) photons cm-2, which is calculated to bleach approximately 20% of the photopigment molecules in green sunfish rod outer segments. This degree of photopigment bleach is approximately 6-7 orders of magnitude greater than that required to elicit half maximal changes in membrane potential in other vertebrate rod preparations. Furthermore, the reciprocal relationship between light pulse intensity and duration in eliciting an equal elongation response held for relatively long light pulse durations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405168 TI - Interactions between forskolin, GS, and divalent cations on ciliary process adenylyl cyclase and intraocular pressure in the rabbit eye. AB - The major adenylyl cyclase activity (AC) of rabbit ocular ciliary processes was investigated by dose-response analysis with respect to interactions of its regulators [the stimulatory G-protein (alpha S) and forskolin] under both saturating and suboptimal divalent cation (M2+) conditions. alpha S was generated directly with fluoroaluminate or via receptors for vasoactive intestinal peptide or isoproterenol. Forskolin and alpha S when liganded simultaneously to AC at suboptimal M2+, potentiated maximal enzyme activity (approximately twice the sum of the separate activities), while the responses were additive (no potentiation) under saturating M2+ conditions. The results demonstrate that the three ligands (forskolin, M2+ and alpha S) mutually interact in a cooperative manner as follows: (1) The apparent k(act) of forskolin for AC was decreased by 1.7-1.9 log units when alpha S was liganded to AC. (2) Conversely, with forskolin liganded to AC the apparent k(act) of alpha S for AC also decreased by a factor of at least 3 4. (3) The nature of the divalent cation M2+ had no effect on the k(act) of alpha S for AC alone and for the AC:FSK complex, and also did not affect the apparent k(act) of forskolin for AC alone, or for the AC: alpha S complex. The nature of M2+ (Mg2+ or Mn2+) affected only the maximal catalytic rate (Emax). (4) Potentiation of adenylyl cyclase by alpha S + FSK was demonstrable in vivo by the increased ocular pressure response of the rabbit eye when a low topical dose of isoproterenol (125 ng per eye) and a subthreshold dose of forskolin (100 micrograms per eye) were given in combination. PMID- 8405169 TI - The reaction of bovine lens alpha A-crystallin with aspirin. AB - Acetylation of the lysines of bovine lens alpha A-crystallin has been examined after 0-48-hr incubations of whole alpha-crystallins in 100 mM aspirin. The alpha A-crystallins were isolated after the incubation, proteolytically digested into peptides which were separated by reversed phase HPLC and analysed by mass spectrometry. For the reaction conditions used in this investigation, acetylated lysyl residues were the principal products. The extent of acetylation was quantified from the intensities of the peaks in the fast atom bombardment mass spectra of the modified and unmodified peptides. The modified lysine containing peptides demonstrated that all seven lysyl residues of alpha A-crystallin reacted with aspirin; the extent of acetylation at each lysyl residue varied. Plots of the extent of acetylation vs. time were used to calculate rate constants for the reaction at each lysyl residue. The rate constant for the acetylation of Lys 166, the most reactive, was about seven times greater than for Lys 88, the least reactive. These rate constants were used to calculate the yield of predicted products for the reaction of alpha-crystallin with therapeutic concentrations of aspirin. Comparison of the yield of acetylated alpha-crystallin with the yield of carbamylated alpha-crystallin that might occur due to renal failure indicates that aspirin is not likely to be an effective inhibitor of cataract due to carbamylation of lysyl residues. PMID- 8405170 TI - Isolation, culture and characterization of bovine choriocapillary endothelial cells. AB - A long-term culture of choriocapillary endothelial cells (CCE) from bovine eyes has been established. We developed a micromanipulative technique with a dissecting microscope for isolation of choriocapillary sheets after a prolonged trypsinization of choroid tissues. We then used a cell sweeping technique to ensure a culture free of nonendothelial cell contamination. The cloned CCE possessed most morphological features and biochemical markers of capillary endothelial cells from other species and other origins. The CCE produced Factor VIII-related antigen and angiotensin-converting enzyme. The CCE phagocytized acetylated low density lipoprotein. The cell cycle time of CCE grown in a medium with sufficient soluble growth factors was dependent upon fibronectin (FN) coating concentrations. At a low concentration of FN (0.5 microgram cm-2) the cell doubling time was 42 hr, whereas at high FN (2 micrograms cm-2) the doubling time was 22 hr. The characterized choriocapillary endothelial cells produced by this method may be useful for studies of choriocapillary angiogenesis under physiological and pathological conditions. PMID- 8405171 TI - Nonenzymatic advanced glycation in the lens membranes. AB - Nonenzymatic advanced glycation was induced in bovine lens membranes. The resulting modified membranes display some changes which were also observed in the advanced glycated crystallins: an increased absorption in the near-UV region, appearance of a strong blue fluorescence (Ex/Em = 370/450 nm), and a subtle change in the near-UV circular dichroism (CD). These changes were also observed in diabetic human lens membranes, but with a less extent in nondiabetic samples. The results indicate that advanced glycation is important in diabetes mellitus, not only to crystallins but also to membrane proteins, both of which generate yellow pigment chromophores that result in lens coloration. PMID- 8405172 TI - In vitro aging of bovine and human retinal pigment epithelium: number and activity of the Na/K ATPase pump. AB - We have previously observed that the density of Na/K ATPase pumps is lower in RPE cells in the posterior pole of both bovine and human eyes. The posterior pole in human eyes includes the macula, a region which is predisposed to aging pathology. In this study we examined the effect of age on the sodium pump using cultures of bovine (bRPE) and human (hRPE) RPE cells that were aged in vitro by repeated passage. The cultures were assayed for cell number, protein, pump density (specific binding of [3H]ouabain) and pump activity (specific uptake of 86Rb) at confluency at each passage. In culture, bRPE had more pumps per cell (3.2 x 10(6)) than hRPE (1.2 x 10(6)), but the bRPE pumps were less active so the pumping capacity per cell was nearly equal. Bovine RPE aged more rapidly in vitro (survived fewer passages) than hRPE. With aging, RPE cells from both species showed declines in cell number at confluency. Pump number and pump activity per cell remained constant. Because cell number declined, the pumping capacity per unit area of confluent epithelium was diminished with culture aging. RPE cell number is known to decline with age in situ, especially in the macula. If Na/K ATPase pump number and activity per RPE cell remain constant with aging in vivo as shown here in vitro, the effective pumping capacity of the RPE per unit area of 'monolayer' would decline in aged eyes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405173 TI - Phosphorus-31 NMR study of the effects of UV on squirrel lenses. AB - In a series of 31P NMR experiments the effects of near-UV radiation on the phosphorus metabolites in pigmented lenses from grey squirrels were studied; UV radiation ranged from 300 to 400 nm, peaking at 365 nm. Squirrels were killed, intact lenses removed, exposed to UV, put in an NMR perfusion cell, and 31P NMR spectra were recorded. In vitro exposure of squirrel lenses for periods ranging from 4 to 22 hr were made. ATP levels, as measured by NMR, decreased as the radiation dose increased. For a 22-hr exposure (121 J cm-2) ATP levels decreased by 67% relative to control values. At lower dose levels there was a correspondingly smaller decrease in ATP levels. Histological examination showed UV-irradiated lenses had disrupted epithelium, loss of attachment to cortical fiber cells and fluid accumulation under the capsule. It is clear that lenticular exposure to near-UV, predominantly at 365 nm (UV-A), does cause metabolic deterioration through degradation of normal ATP energy levels. The grey squirrel pigmented lens strongly absorbs near-UV radiation which enhances the observed metabolic changes and observed opacities. The results demonstrate the utility of using 31P NMR to evaluate metabolic changes associated with near-UV irradiation. PMID- 8405174 TI - A comparison between the effect of topical and systemic carbonic anhydrase inhibitors on aqueous humor secretion. AB - We have assessed the onset and duration of decreased intraocular pressure and aqueous humor flow contrasting systemic and topical administration of carbonic anhydrase inhibitors. The relationship between physiological effects and fractional activity of carbonic anhydrase isoenzymes in the eye was also investigated. Experiments were performed in normotensive New Zealand white rabbits. Intraocular pressure was determined manometrically or tonometrically and aqueous humor flow by sulfacetamide clearance. We studied methazolamide (25 mg kg 1), ethoxzolamide (4 mg kg-1), and MK-927 (2% in 0.5% hydroxyethylcellulose, topical, pH 4.8). There is an immediate reduction in intraocular pressure (1.2 and 1.8 mmHg by 2 min) and aqueous flow (33% and 40% by 5 min) following intravenous dosing with either methazolamide or ethoxzolamide. This correlates with rapid appearance of drug in the anterior uvea and very low fractional activity of ocular carbonic anhydrase isoenzymes II (cytosolic) and IV (membrane bound). Peak intraocular pressure reduction averaged 4.2 +/- 0.68 mmHg and 4.5 +/ 0.8 mmHg for methazolamide and ethoxzolamide at 60 and 45 min, respectively. Peak flow reduction was 38% for methazolamide and 40% for ethoxzolamide, at 5 min. Aqueous flow and intraocular pressure returned to baseline at 7 and 4 hr following methazolamide and ethoxzolamide, respectively. This corresponds to decay of drug from ocular tissues and significant increases in fractional activity of carbonic anhydrase isoenzymes. Topical MK-927 resulted in a 1.2 mmHg decrease in pressure by 5 min. This correlated with the early appearance of drug in the anterior uvea prior to its appearance in aqueous humor and very low fractional activity of carbonic anhydrase isoenzymes. Intraocular pressure decreased 3.6 +/- 0.35 mmHg at 1 hr and returned to baseline by 6 hr. Aqueous flow was reduced 12% by 5 min and 35% at 1 hr. The appearance of MK-927 in the anterior uvea prior to detection in aqueous suggests a significant non-corneal route of absorption following topical administration. Topical MK-927 results in a more gradual reduction in intraocular pressure and flow, although peak effects are not statistically different from systemic carbonic anhydrase inhibitors. The time of pressure return to baseline is also comparable to systemic carbonic anhydrase inhibitors. Because the relations between carbonic anhydrase II and carbonic anhydrase IV in the ciliary process are not yet clear and since the drugs have different affinities for the isozymes, the precise degree of fractional inhibition necessary for pharmacological effect is not certain, but based on drug concentration in the anterior uvea, may take 98% inhibition for full intraocular pressure reduction. PMID- 8405175 TI - Recurrent anterior uveitis induced by multiple systemic injections of muramyl dipeptide. AB - The development of new therapeutic modalities in the treatment of uveitis has been greatly aided by the availability of suitable animal models. The endotoxin model excellently mimics many features of acute anterior uveitis. A new model that exhibits many features of recurrent anterior uveitis is described here. Muramyl dipeptide, MDP, is a synthetic monomer of Gram positive and Gram negative bacterial cell walls. The molecule is highly inflammatory, but unlike endotoxin does not elicit an antibody response. After a single injection (4 hr post injection), acute anterior segment changes include massive engorgement of peri limbal blood vessels, anterior chamber flare, fibrin deposits and heterophil (neutrophil) infiltration which subside within 24-48 hr. There is also break-down of the blood-aqueous barrier. With multiple injections the exacerbations and remissions continue to be observed after each injection but the disease progresses to a recurrent form including the development of synechiae and a mononuclear infiltrate into the anterior segment. The ocular changes are paralleled by changes in the bloodstream with recurrent heterophilia. Thus MDP does not elicit ocular or systemic tolerance. This new model of anterior uveitis should allow study of many of the events that occur in recurrent uveitis in man. PMID- 8405176 TI - Purinergic receptors in ocular ciliary epithelial cells. AB - Adenosine and ATP have been shown to activate separate cell surface purinergic receptors which have been designated P1 for adenosine and P2 for ATP. The pharmacological characterization of P1 and P2 purinergic receptor-mediated signal transduction has been performed in cultured cell lines of the ciliary epithelium. In ODM Clone-2, a cell line derived from human nonpigmented ciliary epithelium (NPE) and in a clone derived from bovine pigmented ciliary epithelium (PE), we observed that adenosine inhibits adenylate cyclase activity at high potency (nM) and stimulates adenylate cyclase activity at low potency (microM) suggesting the presence of P1 subtypes on these cell membranes. The selective agonist cyclopentyladenosine (CPA) was effective at inhibiting forskolin-stimulated adenylate cyclase in these cells. The IC50 for CPA in both NPE and PE was approximately 1 nM in the absence, and 11 nM in the presence of 3-isobutyl-1 methylxanthine (IBMX). In NPE, the selective agonist 2-[p-(2 carboxyethyl)phenethylamino]-5'-N-ethylcarboxamido adenosine (CGS 21680) stimulated adenylyl cyclase with an EC50 of 11 +/- 4 nM in the presence of 4-(3 butoxy-4-methyoxybenzyl)-2-imidazolidinone (RO-20-1724), a phosphodiesterase inhibitor devoid of adenosine receptor antagonism, and 61 +/- 8 microM in the presence of IBMX. In PE cells, EC50 value of RO-20-1724 was 19 +/- 5 nM (n = 3). The characterization of P2 receptors based upon the ability of ATP and its related analogues to stimulate inositol phosphate production reveal the presence of a putative P2u receptor in both cell types. PMID- 8405177 TI - In vitro evaluation of antiproliferative potential of topical cyclooxygenase inhibitors in human Tenon's fibroblasts. AB - Pharmacologic agents which inhibit fibroblast proliferation and scar formation may improve the success of glaucoma filtration surgery. To evaluate the potential use of nonsteroidal anti-inflammatory drugs as an antifibrotic treatment following glaucoma filtration surgery, the time and dose-related effects of four commonly used topical cyclo-oxygenase inhibitors--diclofenac, flurbiprofen, piroxicam, and suprofen--on human Tenon's fibroblast attachment and proliferation were studied. Fibroblasts were incubated with different concentrations of a drug and analysed for dose response 1, 3, and 8 days following drug addition. The cell density was quantified by Coulter counting, hexosaminidase assay, and [3H]thymidine uptake into the DNA. All four drugs inhibited fibroblast attachment at the highest concentrations. Diclofenac was the most effective agent in inhibiting fibroblast proliferation, with inhibition occurring at a range of concentrations above 10(-3)-10(-4) mmol l-1. Flurbiprofen, piroxicam, and suprofen were comparable to each other in their antiproliferative activities, with inhibition occurring at a range of concentrations above 10(-1)-10(-2) mmol l 1. These results suggest a potential role for the commonly used topical nonsteroidal anti-inflammatory drugs in preventing excessive fibroblast proliferation following GFS, but further clinical studies are indicated to assess their activity as anti-inflammatory and antiproliferative agents in preventing bleb failure. PMID- 8405178 TI - L-arginine-derived nitric oxide: a major determinant of uveal blood flow. AB - The effect of inhibition of the L-arginine-nitric oxide pathway on regional ocular blood flow was studied in anaesthetized Beagle dogs. Under control conditions (mean arterial blood pressure 105 +/- 6 mmHg) trapping of 11 and 15 microns-diameter tracer microspheres revealed the following regional blood flows (ml min-1 g-1): retina 0.065 +/- 0.016, choroid 5.72 +/- 0.32, ciliary body 0.77 +/- 0.11, iris 0.18 +/- 0.04. After i.v. infusion of nitro-L-arginine methylester (20 mg kg-1), a potent inhibitor of nitric oxide production from L-arginine, mean arterial blood pressure increased from 105 +/- 6 mmHg by 19% to a new steady state level of 125 +/- 7 mmHg. This increase of arterial blood pressure extended over 3 hr and was reversible after i.v. infusion of L-arginine (100 mg kg-1 over 10 min). Despite the increase in blood pressure following infusion of nitro-L arginine methylester blood flow of choroid, ciliary body, and iris were significantly decreased by 40, 40 and 48%, respectively. Retinal blood flow did not change significantly (-12%). These results suggest that the L-arginine-nitric oxide pathway is of major importance for the adjustment of uveal blood flow under resting physiological conditions. PMID- 8405179 TI - Identification of the alpha-subunits of rod and cone transducin in chicken photoreceptor cells. AB - The chicken retina has several types of cone photoreceptor cells, each of which contains a visual pigment, chicken red (iodopsin), green, blue or violet. Although biochemical and photochemical properties of these cone pigments have been well characterized, no information is available about the chicken photoreceptor G-protein, transducin, which couples with the visual pigment to convert a photon signal into a cellular response. To identify alpha-subunits of chicken rod and cone transducins (Tr alpha and Tc alpha, respectively), we produced two site-directed antibodies which discriminate between bovine Tr alpha and Tc alpha. Immunohistochemical studies on chicken retinas revealed that the antibody against bovine Tr alpha specifically stained the rod outer segments. On the other hand, the antibody against bovine Tc alpha uniformly stained the outer segments of the double cones and all types of single cones, while the single cones were immunohistochemically classified into three types by using a combination of antibodies against bovine rhodopsin and chicken iodopsin. Immuno blot analyses demonstrated that the antibody against Tc alpha recognized a single band of chicken photoreceptor protein, whose molecular weight (42,000) was in good agreement with that of bovine Tc alpha (41,000). The antibody against Tr alpha recognized a protein having the same molecular weight as that of bovine Tr alpha (39,000). These observations strongly suggested that all types of chicken cone cells have a single common Tc alpha (42 kDa) structurally related to bovine Tc alpha, though each cone cell type has a distinct visual pigment. PMID- 8405180 TI - Horizontal cells of the normal and dystrophic rat retina: a wholemount study using immunolabelling for the 28-kDa calcium-binding protein. AB - Immunocytochemistry with a monoclonal antibody against the 28-kDa calcium-binding protein (28-kDa CaBP) was used to study horizontal cells in both congenic control (RCS-rdy+) and dystrophic (RCS) 3-month-old rat retina. We found that horizontal cells in the RCS-rdy+ rat retinas were distributed evenly from the centre to the peripheral retina with a mean density of 783 cells mm-2. Cell size ranged between 6.5-12.5 microns in diameter with a mean of 8.87 microns. Individual cell processes could not be followed readily, however they formed a narrow plexus within the outer plexiform layer with the primary dendrites extending radially from each cell soma in an organized manner. In the RCS rat, by contrast, the horizontal cell processes were more disorganized than in the congenic controls. The degree of disorganization varied across the retina. In the posterior pole, where all outer layers had been lost, many processes were grossly swollen along their length and at the tips. The cell somas were also distributed more widely in the depth of the retina. In the peripheral retina, where a debris layer was still present, the processes were less swollen but they extended abnormally widely in depth. Despite the disorganized structure the mean and range of cell size (9.23 microns, 5.5-15.5 microns), and cell density (796 cells mm-2) were similar to that of the control RCS-rdy+ animals. These findings indicate that in dystrophic retinas beyond a certain stage of degeneration the horizontal cells become abnormal in their structure but are present at a normal density and the somas are not grossly swollen or shrunken.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405181 TI - The modification of proteins by 3-hydroxykynurenine. AB - The autoxidation and reactivity towards proteins of 3-hydroxykynurenine, a tryptophan metabolite found in the human lens, has been studied. At neutral pH, 3 hydroxykynurenine was readily oxidized using molecular oxygen with the formation of several coloured products. The autoxidation of both 3-hydroxykynurenine and the related aminophenol, 3-hydroxyanthranilic acid, was inhibited by the inclusion of sulphydryl compounds such as glutathione or cysteine. Covalent adducts involving the thiols were not observed with either aminophenol. 3 Hydroxykynurenine was found to react with proteins, including lens proteins, to produce brown-coloured polypeptides characterized by an indistinct long wavelength absorption. This protein tanning was inhibited by glutathione. Despite the presence of an amino group in the side chain of 3-hydroxykynurenine, this tanning of proteins was found to involve amino groups including those of lysine residues, as has been found for 3-hydroxyanthranilic acid. Although both aminophenols tanned polylysine, only 3-hydroxykynurenine induced precipitation of the polyamino acid. 3-Hydroxykynurenine tanned all of the purified crystallins but induced precipitation only in the case of alpha A-crystallin. The implications of these findings for senile cataract are discussed. PMID- 8405182 TI - The effect and recovery of long-term H2O2 exposure on lens morphology and biochemistry. AB - Oxidative stress has long been speculated to play an important role in cataractogenesis. In the H2O2-induced cataract model, rat lens showed extensive biochemical damage but very mild morphological changes after being exposed to H2O2 (0.5 mM) for 24 hr in culture. This damage included reduced glutathione (GSH) depletion, protein-GSH mixed disulfide (PSSG) elevation but not protein protein disulfide (PSSP) formation. In order to understand the role of protein thiol mixed disulfide formation in relation to the sequence of events during cataract induction, we conducted a long term H2O2 exposure study for up to 96 hr to monitor the dynamic changes in GSH and PSSG levels, the formation of PSSP aggregate, protein solubility, and the progression in lens opacity. Rat lenses were cultured in 0.5 mM H2O2 and harvested at intervals of 24, 48, 72 and 96 hr for the examination of morphological and biochemical changes. Contralateral lenses cultured in H2O2-free media were used as controls. It was found that the lenses had only patchy opacity at the equator after 24 hr, but became hydrated suddenly at 48 hr (31% heavier than the control), with an opacity which involved the entire outer cortical region. By 72 hr incubation, the nucleus was opacified. Lens GSH progressively decreased with time of H2O2 exposure, 40% was lost by 24 hr and over 95% by 48 hr. There was a concomitant elevation of PSSG, 16-fold over the controls by 24 hr and 45-fold by 48 hr followed by a decline to 34-fold after 72 hr. In addition, the level of protein-cysteine mixed disulfide (PSSC) was elevated after 48 hr incubation in H2O2. At this time point, PSSP aggregates began to appear both in water soluble (WS) and urea soluble (US) fractions along with a drastic reduction in protein solubility. Western blot analysis of the protein fractions identified beta and gamma, but not alpha-crystallin in the disulfide-containing aggregates. The lens clarity and biochemical changes partially recovered if the oxidant was removed within 24 hr, indicating a potential therapeutic role for antioxidants. The complete normalization of PSSG level under this recovery condition signifies that cells may have a natural defense system for controlling PSSG elevation. PMID- 8405183 TI - Immunopathology of reactivation of experimental ocular histoplasmosis. AB - We have established a non-human primate model of experimental ocular histoplasmosis. This model has been shown to result in chronic lesions that resemble typical 'histo spots' or choroidal scars but that contain infiltrates of lymphocytes for as long as 10 yr following intracarotid injection of live Histoplasma capsulatum. Using this model, we attempted to reactive these late choroidal lesions via intracarotid challenge with specific antigen (heat-killed H. capsulatum). No clinical changes suggestive of reactivation of these lesions were observed following this antigenic challenge. However, immunopathologic analysis of choroidal lesions at 1, 3 and 7 days after antigenic challenge revealed significant increases in both the numbers of inflammatory cells and the relative percentages of the helper/inducer lymphocyte and macrophage populations. Our results demonstrate that, following antigenic challenge, a cellular change, consistent with a type IV delayed hypersensitivity, can be observed in previously active, but clinically quiescent, histoplasmosis lesions. In light of the many parallels between our primate experimental model and human ocular histoplasmosis, our findings suggest that, in the human, significant immunopathologic activity may occur subclinically in the choroid of affected individuals. It is possible that repeated bouts of subclinical reactivation may induce or enhance chronic choroiditis and, over many years, ultimately produce slow progressive damage to the Bruch's membrane/retinal pigment epithelium complex, resulting in clinically 'active' macular disease and, in selected cases, subretinal neovascularization. PMID- 8405184 TI - Increased expression of chloramphenicol acetyltransferase by carotenoid and retinoid replacement in Drosophila opsin promoter fusion stocks. AB - Drosophila promoter fusion stocks containing a chloramphenicol acetyltransferase (CAT) reporter gene fused to a 2.8 kb DNA fragment from the Rh1 opsin promoter were carotenoid deprived from egg to adult, and then adults were replaced by feeding carrot juice. CAT activity, determined by radiometric assay, was low in deprived flies; it increased rapidly during the first 3 days of replacement and then declined back to the control level. Retinoic acid increased peak CAT activity as much as carrot juice and more than beta-carotene, all-trans retinol or all-trans retinal. These findings suggest that vitamin A serves not only as rhodopsin's chromophore but also influences Rh1 opsin gene transcription. Three stocks with various deletions in the Rh1 opsin promoter lacked the carrot juice dependent elevation of CAT activity. All three deletions include the region from 701 to -488, suggesting that this region may contain a vitamin A-responsive DNA sequence. PMID- 8405185 TI - Developmental appearance, species and tissue specificity of mouse 23-kDa, a retinal calcium-binding protein (recoverin). AB - A 23-kDa, soluble, calcium-binding photoreceptor-specific protein (23-kDa) has been shown to be identical to recoverin and the cancer-associated retinopathy protein. Recoverin has been reported to activate guanylate cyclase to increase the amount of cyclic GMP and thereby reopen cation channels within the photoreceptor cells. In this study, the 23-kDa protein was purified from bovine retinas and monospecific antibodies against it were generated in rabbits. Western analysis demonstrated 23-kDa in retinas from human, monkey, bovine, dog, rabbit, rat, mouse, frog, chameleon and iguana although it was not detected in chicken or fly retinas. No immunoreactivity was observed in any non-retinal tissues except the pineal gland. The 23-kDa protein was detected, by Western analysis, at postnatal day 5 in the mouse retina and it increased in amount in parallel with the differentiation of the photoreceptor cells in normal mice and it also decreased in parallel with their degeneration in the rd mouse. Immunocytochemical analysis of the adult mouse retina showed that 23-kDa is restricted primarily to the inner segments of the photoreceptor cells and, unlike arrestin, its localization did not shift in response to light/dark changes. PMID- 8405186 TI - Structural characterization of lipid membranes from clear and cataractous human lenses. AB - Lipid composition related structural changes in human cataractous lenses was explored by characterizing the hydrophobic hydrocarbon chains in lipid membranes corresponding to twelve Indian cataractous lenses and eight American clear lenses of similar age. The nuclear-lipid phase transitions corresponding to the clear lenses exhibited significantly higher average transition temperatures (nucleus 33 degrees C, cortex 26.3 degrees C) and cooperativities, 38.1, as compared to the value of 24.1 for the cortical-lipid phase transitions. At 36 degrees C, the phase transitions corresponding to cortical and nuclear lipids indicate a similar degree of disorder, 63%, in the hydrocarbon chains, i.e., similar relative amounts of gauche and trans rotomers. The twelve cataractous lenses investigated all had nuclear opacities, four were brunescent and four had cortical opacities. No significant differences were observed in the phase transition parameters (temperature, cooperativity, magnitude, enthalpy) evaluated for the nuclear-lipid membranes corresponding to the different types of cataracts. Furthermore, for the cataractous membranes, the phase transition parameters obtained for the nuclear lipids were comparable to those evaluated for the cortical lipid membranes. However, the cortical lipids exhibited the highest order in membranes from nuclear cataracts without cortical opacity. The cortical lipids from clear, non cataractous lenses had the lowest level of order. At 36 degrees C, the degree of order in the cortical lipid from clear lenses was comparable to that from nuclear cataractous lenses without cortical opacity. The transition temperature, and cooperativity were significantly higher for cortical lipids from cataractous lenses as compared to those from clear lenses. At 36 degrees C, the degree of order in the cortical lipid membranes was lower for all cataract types vs. clear lens fractions. Our results suggest the possibility that lipid-lipid interactions could be different in cataractous lens membranes. Lipid compositional and chemical differences must account for these altered lipid interactions. These studies will provide a basis for studying lipid-protein interactions and structure-function relationships in the lens membrane. PMID- 8405187 TI - The presence of serotonin (5-HT1) receptors negatively coupled to adenylate cyclase in rabbit and human iris-ciliary processes. AB - Serotonin (5-hydroxytryptamine, 5-HT) reduces forskolin-induced stimulation of cyclic AMP in rabbit iris-ciliary body (ICB) homogenates. The effect is dose dependent and can be mimicked by a number of 5-HT1 receptor agonists including 5 carboxamidotryptamine (5-CT) and RU 24969 [5-methoxy-3-(1,2,3,6, tetrahydro-4 pyridinyl)-1-indole]. The inhibitory effects of 5-CT and the 5-HT1A selective agent 8-hydroxy-2-(di-n-propyl-amino) tetralin (8-OH-DPAT) on forskolin stimulated adenylate cyclase activity are greater in isolated ciliary processes than in the iris musculature. Spiperone and propranolol significantly antagonize the action of 5-CT in the iris-ciliary body, while ketanserin (5-HT2 antagonist) and ICS 205930 (5-HT3/4 blocker) were without influence, indicating the presence of the 5-HT1A subtype of receptor. Studies carried out on human ICB homogenates also suggest the presence of 5-HT1A-like receptors, although these receptors are not identical to those in rabbit. Similarities include dose-dependent decreases in cAMP levels stimulated by forskolin elicited by 1-(3-chlorophenyl) piperazine (mCPP), 5-CT and 8-OH-DPAT. Moreover, the inhibitory effect of 5-CT can also be significantly reduced by the 5-HT1 receptor antagonist, propranolol. However, unlike the case of rabbit tissue, spiperone was ineffective in abolishing the 5 CT response in human ICB homogenates. PMID- 8405188 TI - Enzyme activities and crystallin profiles of clear and cataractous lenses of the RCS rat. AB - Royal College of Surgeons (RCS) rats have hereditary retinal degeneration in association with posterior subcapsular opacities. Cataract formation is thought to be correlated with an increase in lipid peroxidation products in the vitreous (Zigler and Hess, 1985). In order to examine the possibility that parallel changes in enzyme activity are occurring within the lens, we analysed the activity of four key enzymes and the crystallin protein profile. We compared RCS rat lenses at three different stages of cataract formation to clear lenses of the nonpigmented RCS rat, lenses from pigmented RCS rat and to normal (Fisher) rat. Our data shows that concomitant with the appearance of the RCS cataract, the ratio of the crystallins beta 1, beta H and gamma to the total lens protein was reduced. The crystallin profile of a clear RCS lens was similar to that of a normal (Fisher) lens. No significant difference in the activity of the enzymes hexokinase and glucose-6-phosphate dehydrogenase (G6PD) was found among the lenses, however the activity of glutathione reductase and aldolase was reduced in the cataractous lenses. PMID- 8405189 TI - Transport of 22:6n-3 in the plasma and uptake into retinal pigment epithelium and retina. AB - Vertebrate retina is highly enriched in 22-carbon polyunsaturated fatty acids (PUFAs), especially docosahexaenoic acid (DHA, 22:6n-3), which is supplied by the plasma. To study the transport and uptake of 22:6n-3 into the retina, young Long Evans rats were fasted overnight and given an oral dose of palm oil containing [4,5-3H] 22:6n-3 and [9,10-3H] 18:1n-9 (oleic acid). At 1, 6, 12, 24, and 48 hr postgavage, blood, liver, retinal pigment epithelium (RPE) and retina were collected, and plasma lipoprotein fractions were obtained by density-gradient ultracentrifugation. The distribution of radioactivity among the different lipoproteins (dpm%) showed that both fatty acids were mainly associated with VLDL 1 hr after gavage. For 22:6n-3, the majority of radioactivity was associated with LDL at 6 hr postgavage. By 12 hr post administration, the majority of radioactivity was in HDL2 which retained around 40% until the 48-hr time point. For 18:1n-9, a different profile was observed. By 6 hr postgavage, approximately 30% of the radioactivity was found in HDL2, but the percentage of radioactivity decreased to approximately 20% thereafter. The half-life of 22:6n-3 was significantly longer than for 18:1n-9 (P < 0.0005) in plasma, liver, and the lipoprotein fractions. The specific activities of 22:6n-3 and 18:1n-9 in the plasma continuously declined from 1 hr to 48 hr postgavage, while liver showed a peak of specific activity at 6 hr, then gradually declined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405190 TI - Aldose reductase in human retinal pigment epithelial cells. AB - Sugar alcohols have been reported to accumulate in retinal pigment epithelium (RPE) of diabetic animals. This finding has raised interest in the role of RPE in diabetes-associated retinal changes such as cystoid macular edema. To confirm the presence of aldose reductase in this tissue, the NADPH-dependent enzyme was purified to an apparent homogeneity from cultured human RPE cells, characterized, and its biochemical properties investigated. The induction of aldose reductase by hypertonic stress was also examined. The purification of aldose reductase was performed by a series of chromatographic steps which include gel filtration, affinity chromatography and chromatofocusing. Final purity achieved was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The kinetic properties and susceptibility to inhibition of the purified aldose reductase were essentially identical to aldose reductase purified from human placenta and kidney. In addition to aldose reductase, chromatofocusing demonstrated the presence of aldehyde reductase, another NADPH-dependent reductase. However, the amounts of aldehyde reductase present were much smaller than those of aldose reductase and the levels of aldehyde reductase appeared too small to contribute to the polyol production in the RPE cells. Culture of RPE cells in hypertonic medium containing 150 mM sodium chloride (600 mosmol total) increased both reductase activity, monitored with DL-glyceraldehyde as substrate, and immunoblot staining for aldose reductase. Chromatofocusing of RPE cells cultured in hypertonic media resulted in a prominent increase in the peak corresponding to aldose reductase compared to the peak height of cells grown in control medium. No increase in aldehyde reductase from RPE cells cultured in hypertonic medium was observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405191 TI - Low and high Km forms of dinitrophenylglutathione-stimulated ATPase in bovine lens. AB - Dinitrophenyl S-glutathione (Dnp-SG) ATPase which catalyses the hydrolysis of ATP in the presence of GSH-conjugates has been implicated previously in the transport of these conjugates. In the present studies we demonstrate that Dnp-SG ATPase is present in bovine lens epithelium and cortex. The specific activity per mg membrane protein was found to be 75-fold higher in the epithelium as compared to the cortex. No enzyme was detected in the nuclear region of the lens. Dnp-SG ATPase was purified from bovine lens epithelium and cortex using Dnp-SG-Sepharose 6MB affinity chromatography. The partially purified Dnp-SG ATPase had two distinct Km values, 120 microM and 1.0 mM. The antibodies raised against human erythrocyte Dnp-SG ATPase cross-reacted with the bovine lens epithelium Dnp-SG ATPase which was identified by Western blot as a band corresponding to an approximate M(r) value of 80,000 Da. PMID- 8405192 TI - Preparative scale isolation and partial purification of human rod outer segments. PMID- 8405193 TI - Isolation and characterization of a cDNA for mouse retinal phosducin. PMID- 8405194 TI - Diethylstilbestrol (DES) and breast cancer. PMID- 8405195 TI - Diet, body size, and breast cancer. PMID- 8405196 TI - Alcohol consumption and risk of breast cancer: a review of the epidemiologic evidence. PMID- 8405197 TI - Cigarette smoking and the risk of breast cancer. PMID- 8405198 TI - Radiation and other environmental exposures and breast cancer. PMID- 8405199 TI - Recent etiologic hypotheses concerning breast cancer. AB - A few studies have noted moderate elevation in the risk of breast cancer among women with residential exposure to electromagnetic fields, among women without a history of much strenuous physical activity, and among women with in utero exposures that may indicate high levels of maternal estrogen. The relative risk for each of these associations has generally been less than 2, with little adjustment for possible confounding factors. Also, several studies have not been able to confirm these relations. Currently there is scant or no evidence that silicone breast implants or psychological factors increase the risk of breast cancer. Despite the limited number of studies, the inconsistent results, and the difficulty of measuring several of these exposures, the plausible biologic mechanisms for each indicate that more research on these hypotheses is warranted. PMID- 8405200 TI - Multiple primary cancers involving the breast. PMID- 8405201 TI - Estrogens, progestogens, normal breast cell proliferation, and breast cancer risk. PMID- 8405202 TI - Benign breast diseases, carcinoma in situ, and breast cancer risk. PMID- 8405203 TI - Nipple aspirate fluid in epidemiologic studies of breast disease. PMID- 8405204 TI - Mammographic parenchymal patterns: a marker of breast cancer risk. AB - There is now a large amount of evidence showing that mammographic densities are an indicator of increased risk of breast cancer. There is as yet no generally agreed upon and recognized method of classifying these densities, although the available evidence shows that quantitative description of densities creates larger gradients of risk than Wolfe's classification and larger risk gradients than most other risk factors for breast cancer. It seems likely that improved methods of describing densities quantitatively, and possibly other methods of characterizing the tissue changes that are responsible for the densities, will allow greater discrimination. However, it is already clear that breast cancer develops in a large number of women who do not have radiologic changes indicating increased risk, and that it is unlikely that mammographic pattern, or any other risk factor for breast cancer identified to date, will be useful for the selection of women for mammographic screening. Although mammographic densities are associated with an increased risk of developing histologic changes that are risk factors for breast cancer, the histologic feature most consistently associated with mammographic densities is stromal fibrosis. We suggest that the relation between stromal fibrosis and risk of breast cancer can be explained by the known actions of a variety of growth factors that are thought to play a role in a number of aspects of breast development and carcinogenesis. The association between mammographic densities and several other risk factors for breast cancer suggests that these factors may also modulate the activity of growth factors in breast tissue, and that this may be the means by which they influence breast cancer risk. Further research is needed to determine whether differences in the activity of growth factors in breast tissue can be found in association with radiologic and other risk factors for breast cancer. The available evidence indicates, therefore, that mammographic parenchymal patterns do, at least in part, meet the criteria outlined in the introduction of this paper. Some mammographic appearances are associated with a substantial increase in the risk of breast cancer, and, as shown by observations on the effects of hormone use, are capable of change. Mammographic densities have also been found to be associated with biochemical characteristics of possible relevance to carcinogenesis. The appearances that are related to risk may, therefore, be most useful as a means of investigating the etiology of breast cancer and of testing hypotheses about potential preventive strategies. PMID- 8405205 TI - Hormone receptors and breast cancer. PMID- 8405206 TI - Breast cancer in men. AB - Epidemiologic studies of breast cancer in men have provided insights into the pathogenesis and etiology of breast cancer in both sexes. Individual carcinomas from both the male and female breast are histologically indistinguishable, but histologic types of ductal origin occur relatively more frequently in men than in women, and those of lobular origin are very uncommon in men, reflecting the absence of lobular structures in the normal male breast. The same variations in incidence and mortality rates of breast cancer among countries and racial and ethnic groups that have been observed in women also occur in men, clearly indicating that the causes of these variations are not primarily risk factors related to being female. Risk of breast cancer in men increases with age, with no change in the rate of increase at the usual age of menopause; this supports the assumption that the midlife change in the rate of increase with age in women is due to the reduction in ovarian hormone production at menopause. Incidence rates of breast cancer in men have remained stable over time, suggesting that the temporal increase in rates in women is a result of either enhanced detection due to screening or changes in risk factors that are sex-specific. In men, an increase in risk of breast cancer has been associated with testicular pathology and dysfunction, and a decrease in risk has been related to high fertility, a history of prostate cancer, and exogenous androgens. These observations suggest that risk may be enhanced by low levels, and reduced by high levels, of androgens. Conversely, high estrogen levels probably increase risk of breast cancer in men, since risk has been associated with several conditions that may result in hyperestrogenism. These conditions included obesity, rapid weight gain, elevated blood cholesterol, gallstones, non-insulin-dependent diabetes, and chronic liver diseases. Studies of the role of endogenous hormones in the etiology of breast cancer in women have tended to focus on estrogens (71). These observations on breast cancer in men suggest that the relative levels of androgens and estrogens may be of etiologic importance, and that additional studies in women should include measurements of androgens. A history of breast cancer in a first-degree relative is associated with about a doubling of the risk of breast cancer in both men and women.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405207 TI - Tamoxifen for the primary prevention of breast cancer: a review and critique of the concept and trial. AB - This paper has reviewed the rationale for and design of the NSABP Breast Cancer Prevention Trial and has provided a brief critique of the philosophy of chemoprevention of breast cancer and of certain practical aspects of the trial design. If the assumptions and estimates from the trial protocol are correct, the net benefit of the trial will be moderately large and positive (50 to 77 events prevented). However, over 500 women per year will be treated with tamoxifen unnecessarily. Recalculation of the net-benefit table using another set of reasonable assumptions regarding risks and the trial assumptions regarding benefits shows a negative effect to a small positive effect overall (-17 to 8 events). If the probability of adverse ocular events is included in the net benefit equations, it appears that more harm than good will result from the intervention. In the face of the uncertainty regarding the net benefit of the trial, ranges of these risks and benefits should be provided to both potential and enrolled participants. The lack of significant benefit to participants seen with the recalculations may raise the question of whether the trial should continue as designed. One option would be to limit trial participation to postmenopausal women only, since 1) breast cancer is more common in postmenopausal women, 2) tamoxifen is more effective in postmenopausal women, 3) cardiovascular disease is more common in postmenopausal women, and 4) reductions in cholesterol levels and preservation of bone mass have only been documented in postmenopausal women (11, 27). Even in this case, however, the fundamental philosophical question of whether large numbers of healthy women should be "treated" with a toxic drug for the primary prevention of a rare event remains. PMID- 8405208 TI - Screening for cancer of the breast. PMID- 8405209 TI - Breast cancer epidemiology: summary and future directions. PMID- 8405210 TI - Tribute to Alan S. Morrison. PMID- 8405211 TI - Reproductive factors and breast cancer. PMID- 8405212 TI - Endogenous hormones and breast cancer risk. AB - There is substantial evidence that high estrogen levels in postmenopausal women are associated with an increase in breast cancer risk, but such a relation has not yet been established in premenopausal women, despite biologic evidence that breast epithelial cell division rates are high during the luteal phase of the menstrual cycle when estradiol and progesterone levels are high. The lack of total consistency among studies that have assessed estrogen differences, whether in breast cancer patients versus controls or in subgroups of the population characterized by different risk profiles for breast cancer, is not unexpected given the extraordinarily complex methodological issues that must be addressed in these studies. There has been a clear evolution over time in the level of sophistication of these types of studies, further decreasing the likelihood of finding consistent patterns in the literature. Other hormones may play an important role in breast cancer development as well. Experimental data are particularly compelling for a role of progesterone and prolactin, but hormonal studies in women are not entirely convincing regarding the role of these two hormones, nor is the literature nearly as extensive as it is for the estrogens. Studies of various androgens are even less consistent. Moreover, such studies suffer from a lack of precise hypotheses regarding how these hormones might directly alter risk. PMID- 8405213 TI - Estrogen replacement therapy and breast cancer risk. AB - Assessment of the effects of menopausal estrogens on the risk of breast cancer is obviously complex, with proper evaluations requiring attention to the effects of selection, recall, and surveillance biases; confounding factors; detailed exposure relations; and subgroup variations. Although much discussion has ensued over the extent to which variations between studies might be due to a variety of methodological differences, it is likely that the limited statistical power of many studies may be largely responsible for the divergent effects observed, particularly with regard to assessing the effect of long-term usage. Thus, in case-control studies in which population controls have been used and for which there have been a sufficient number of women using estrogens for approximately 10 or more years, it appears as though such an exposure may be associated with some elevated risk, possibly on the order of 30-80 percent (16, 34, 36-38)--a magnitude of risk that is obviously extremely difficult to detect in most epidemiologic studies. Evaluation of risks associated with short-term use is even more difficult, which has led most investigators to conclude that usage of less than 5 years is associated with no detectable excess risk. Problems with statistical power also plague evaluations of more detailed measures of use, including dosage and type of preparations, although there is some indication that nonconjugated preparations may be associated with a higher risk than other types of estrogens (16, 20, 36, 44, 51). In addition, there is some evidence that recent use may be associated with a modest elevation in risk (32, 33, 53), although effects of surveillance bias are difficult to resolve. Interpretation of the effect is complicated by several studies which have found estrogens to be associated with an increased risk of breast cancer incidence but a decreased risk of death, with the mortality ratios ranging from approximately 0.5 to 0.8 (33, 92, 93). Although one of these studies hypothesized that the relation with mortality may reflect a tendency for women with clinically recognized breast cancers and those at high risk not to be prescribed estrogens for long periods of time, studies showing that estrogen users have more favorable tumor characteristics at diagnosis than nonusers (20, 36) suggest that screening biases may be involved. The numerous contradictions regarding effects of menopausal estrogens from observational studies have heightened interest in the results of several ongoing controlled clinical trials.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405214 TI - Breast cancer: magnitude of the problem and descriptive epidemiology. AB - Breast cancer is the most common cancer among women in the United States. Knowledge of the descriptive epidemiology of breast cancer is useful both in suggesting etiologic hypotheses and, if preventive measures can be identified, in delineating high-risk groups to be targeted for preventive efforts. Demographic risk factors include increasing age (in Western countries), being white for breast cancer diagnosed at age 45 years or more, being black for breast cancer diagnosed at less than 40 years of age, high socioeconomic status, having never married, being of the Jewish faith, urban residence, and residence in the northern (as compared with the southern) United States. Incidence rates are generally highest in North American and Northern European countries, intermediate in Southern and Eastern European and South American countries, and lowest in Asia and Africa. The most notable characteristic of the descriptive epidemiology of breast cancer in recent years is perhaps the rapidly increasing incidence rates in developing countries. Identification of specific reasons for these increasing rates would contribute substantially to our understanding of the epidemiology of breast cancer. PMID- 8405215 TI - Oral contraceptives in relation to breast cancer. PMID- 8405216 TI - Exogenous progestins and breast cancer. AB - More research on the effect of exogenous progestins on breast cancer risk is clearly needed. Biologic evidence that progestins may act synergistically with estrogen to enhance proliferation of breast epithelial cells emphasizes the importance of further exploration of this issue, particularly given the increasing prevalence of exposure to contraceptive and noncontraceptive progestins. No specific type or dose of progestin in monophasic combination oral contraceptives has been linked to breast cancer. Based on the few epidemiologic studies of progestin-only oral contraceptives, there also is no evidence that they increase risk of breast cancer. Two studies found that longer-term use of progestin-only pills was associated with a decreased risk of breast cancer. However, given the low prevalence of use of minipills, it is unlikely that this exposure substantially affects the incidence of breast cancer in the population as a whole. Use of the injectable contraceptive DMPA has been positively associated with risk of breast cancer in some subgroups of women, although no significant overall adverse effect has been observed in the two largest studies conducted to date. There is suggestive evidence that use at an early age or prior to a first term birth and recent use may increase risk of breast cancer. It remains unclear, however, whether or not surveillance bias may explain the positive association observed in recent users. Additional research on DMPA and breast cancer incidence is needed, since studies published to date have lacked sufficient power to evaluate risk in relation to long-term use. Future studies of breast cancer in relation to use of other long-acting progestational agents such as Norplant will also be important. There is concern about the relation between breast cancer incidence and use of combined estrogen-progestin replacement therapy, especially extended periods of use. At the present time, only one study (45) has estimated risk according to duration of use, and it remains uncertain whether the reported elevation in risk seen in long-term users of combined therapy is due to enhanced surveillance for breast cancer among exposed women. Further research will be required before any definitive conclusions can be reached about the potential effect of estrogen-progestin replacement regimens on breast cancer incidence. Future studies should attempt to determine the circumstances under which progestins may alter risk of breast cancer and whether such effects vary by duration of use, dosage, type of preparation, concomitant use of estrogens, regimen of exposure, and the timing of progestin use in relation to age and menstrual and reproductive events.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405217 TI - The stochastic blueness of leukemic stem cells in experimental antineoplastic therapy. PMID- 8405218 TI - The extent of anemia in Belgrade (b/b) rat. PMID- 8405219 TI - Diverse signaling pathways rendezvous at mitogen-activated protein kinase. PMID- 8405220 TI - Control of hematopoietic cell growth by somatomedins. PMID- 8405221 TI - T cell receptor genes: a glance at normal and malignant hematopoiesis. PMID- 8405222 TI - Cell cycle arrest of human hematopoietic progenitors induced by medroxyprogesterone acetate. AB - Clinical studies have shown that continuous or intermittent scheduled administration of medroxyprogesterone acetate (MPA) reduces the bone marrow toxicity induced by antitumor drugs. This MPA myeloprotection has been attributed to an arrest of hematopoietic progenitors in a quiescent phase, although no in vitro studies have demonstrated such an effect of MPA. Human bone marrow cells were preincubated for 3 days with MPA (100 ng/mL) and then exposed to sublethal doses of adriamycin; LD50 was significantly increased in MPA-preincubated cells (896 +/- 172 ng/mL) vs. control cells (162 +/- 37 ng/mL); this protective effect of MPA was shown to be more efficient against S-phase-specific drugs such as 5 fluorouracil (5-FU) than against non-phase-specific drugs such as cisplatin. MPA did not protect several human leukemic cell lines from the cytotoxic action of adriamycin. "Suicide" assays showed that the percentage of myeloid progenitor cells (granulocyte-macrophage colony-forming units [CFU-GM]) in S-phase was significantly reduced from 67 +/- 2.5% (control cells) to 38 +/- 5.5% (24-hour MPA-preincubated cells). These results demonstrate in vitro that MPA exerts a cell cycle arrest of hematopoietic precursors, protecting them from the toxicity of chemotherapy. PMID- 8405223 TI - Effect of bone marrow T lymphocytes treated with CAMPATH 1G on megakaryocyte colony formation. AB - Specific populations of lymphocytes play a significant role in the regulation of megakaryocyte (MK) development. CAMPATH 1G (C1G) and CAMPATH 1M (C1M) are T lymphocyte (TL)-specific monoclonal antibodies (MABs) that are routinely employed to reduce graft-vs.-host disease (GVHD) in allogeneic bone marrow transplantation (BMT). Following BMT, engraftment of the erythroid and myeloid lineages is prompt, but prolonged thrombocytopenia often prevails. We therefore studied the effect on MK colony formation of treating donor bone marrow (BM) with the CAMPATH MABs. MK colonies were grown in plasma clots using postirradiation aplastic canine serum (PICS-J) as MK colony-stimulating factor (MK-CSF). C1M, which causes TL destruction by complement-dependent lysis, had no effect on MK cloning efficiency. C1G is not lytic but causes the elimination of TL in the BMT recipients via antibody-dependent cell cytotoxicity (ADCC). Treatment of donor BM with C1G significantly enhanced the number of early burst-forming units (BFU-MK) and late colony-forming units (CFU-MK) and had no effect on granulocyte macrophage (CFU-GM) or erythroid (BFU-E) colonies. Enhancement of MK cloning efficiency was concentration-dependent between 0.03 and 3 micrograms MAB/10(6) BM mononuclear cells (MNC). Similar results were observed when C1G-treated TL or purified CD4+ TL were co-cultured with untreated autologous BMMNC or peripheral blood (PB) MNC. Conditioned medium (CM) from C1G-treated TL and CD4+ TL contained soluble factors that, when combined with suboptimal doses of PICS-J, potentiated MK growth. C1G in combination with PICS-J also stimulated TL proliferation in a dose-dependent manner. The T cell CM did not contain elevated levels of interleukin-3 (IL-3), IL-6, IL-1 beta, or GM-CSF. Our data provide additional evidence for the involvement of activated TL, and perhaps novel soluble T cell products, in the immunomodulation of megakaryocytopoiesis. PMID- 8405224 TI - Glycosylated insulin-like growth factor II promoted expansion of granulocyte macrophage colony-forming cells in serum-deprived liquid cultures of human peripheral blood cells. AB - This report presents the results of studies investigating the effect of a glycosylated form of insulin-like growth factor II with an apparent molecular weight of 15,000 (appM(r) = 15K IGF-II) and one with a molecular weight of 7500 (M(r) = 7.5K IGF-II) on the expansion of granulocyte-macrophage colony-forming cells (GM-CFC) in human peripheral blood cells. Blood cells were enriched for GM CFC and other CFCs, and liquid cultures of these cells were established in serum deprived medium supplemented with either interleukin-3 (IL-3) alone (no insulin or IGF added to the medium) or with IL-3 plus M(r) = 7.5K recombinant (r) IGF-II or appM(r) = 15K IGF-II. After incubation for 3 days or 1 week, the blood cells were subcultured in plasma clots, and the number of colonies detected at 7 days (D7) and at 14 days (D14) was used to calculate the number of D7 GM-CFC and D14 GM-CFC. The number of GM-CFC in liquid cultures of blood cells incubated for 3 days with IL-3 alone was similar to the number added at day 0. By 1 week, the number of D14 GM-CFC and D7 GM-CFC had increased to 3.5 +/- 0.9-fold (p = .03) and two- to 50-fold (p = .04) of the number at day 3, respectively. There were 1.5- to six-fold more D7 GM-CFC in cultures of blood cells incubated for 1 week with IL-3 plus either 100 ng/mL M(r) = 7.5K IGF-II or 200 ng/mL appM(r) = 15K IGF II than after incubation with IL-3 alone. appM(r) = 15K IGF-II also promoted a two-fold increase in the number of D14 GM-CFC. appM(r) = 15K IGF-II promoted a greater increase in D14 GM-CFC than incubation with IL-3 alone even for blood samples in which M(r) = 7.5K IGF-II did not promote such an increase. The results of these studies demonstrate that physiologic concentrations of appM(r) = 15K IGF II and M(r) = 7.5K IGF-II increased the number of GM-CFC more than IL-3 alone and suggest that appM(r) = 15K IGF-II was more potent than M(r) = 7.5K IGF-II in augmenting IL-3-induced expansion of GM-CFC in serum-deprived liquid cultures of peripheral blood cells. PMID- 8405225 TI - Peripheral blood stem cell acquisition by large-volume leukapheresis in growth factor-stimulated and unstimulated rhesus monkeys: development of an animal model. AB - Twelve (eight unstimulated [UNS], four growth factor-stimulated [ST]) anesthetized adult male rhesus monkeys underwent a single large-volume leukapheresis (> 3 blood volumes processed) in an attempt to define an animal model for use in future peripheral blood stem cell (PBSC) transplantation studies. The cell separator was primed with autologous blood and saline and set according to the manufacturer's mononuclear cell (MNC) protocol using the granulocyte separation and small volume collection chambers with additional modifications. Stimulated animals received 5 micrograms/kg recombinant human interleukin-3 (rhIL-3) on days -12 to -5, 5 micrograms/kg granulocyte-macrophage colony-stimulating factor (GM-CSF) on days -4 to -1, and large-volume leukapheresis on day 0. UNS animals did not receive growth factors. Vascular access was via a triple lumen intra-aortic catheter; blood pressure was monitored via the third lumen. Pre- and post-apheresis blood counts were determined and product hematocrit (Hct), MNC, colony-forming units-granulocyte/macrophage (CFU GM), CD34+, and lymphocyte subsets were studied. During the 100-minute large volume leukapheresis with mean flow rate 26.4 +/- 3.9 mL/min, the pre- and post Hct were 36.6 +/- 2.6 and 32.3 +/- 7.0%, platelets 447 +/- 305 and 154 +/- 77 x 10(9)/L, and MNC 2.7 +/- 0.9 and 1.9 +/- 1.4 x 10(9)/L (all p < .05) in UNS animals. In ST animals, the pre- and post-Hct were 39.0 +/- 5.6 and 34.9 +/- 3.7%, platelets 507 +/- 100 and 150 +/- 9 x 10(9)/L (p < .05), and MNC 4.9 +/- 1.6 and 2.8 +/- 0.7 x 10(9)/L (p < .05). The product contained 98.5 +/- 1.4% MNC, 4.1 +/- 4.1% Hct, 1.9 +/- 0.6 x 10(9) MNC, 9.2 +/- 7.3 x 10(4) CFU-GM, and 3.5 +/ 2.1 x 10(6) CD34+ cells in UNS animals. In ST monkeys, the product contained 49.5 +/- 32% MNC, 6.4 +/- 4.4% Hct, 3.6 +/- 1.8 x 10(9) MNC, 49.3 +/- 39 x 10(4) CFU-GM, and 9.1 +/- 5.5 x 10(6) CD34+ cells. Greater than 75% of the product MNC were CD2+ T cells in ST and UNS animals. Large-volume leukapheresis in rhesus monkeys was tolerated well. Herein we characterize an animal model for large volume leukapheresis in UNS monkeys that is similar to that of human PBSC leukapheresis. In ST animals there is more than a five-fold increase in CFU-GM collected and an increase in circulating CFU-GM/MNC.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405226 TI - Interleukin-3 and p210 BCR/ABL activate both unique and overlapping pathways of signal transduction in a factor-dependent myeloid cell line. AB - Chronic myelogenous leukemia (CML) is characterized by a specific chromosomal translocation occurring between the long arms of chromosomes 9 and 22 resulting in a fusion product, p210 BCR/ABL, which has elevated tyrosine kinase activity. Expression of p210 BCR/ABL in murine interleukin-3 (IL-3)--dependent cell lines typically converts these cell lines to factor-independence by a non-autocrine mechanism. The IL-3 receptor is believed to function in part by activating a receptor-associated tyrosine kinase, leading to the hypothesis that p210 BCR/ABL may induce factor-independence of myeloid cells by constitutively phosphorylating some common signal-transducing proteins that normally would be phosphorylated on tyrosine residues in response to IL-3. p210 BCR/ABL subclones were constructed from an IL-3-dependent murine myeloid cell line, 32Dcl3, by transfection of a plasmid containing a full-length p210 BCR/ABL cDNA. Following transfection, the cells became completely factor-independent within 3 weeks. We examined the effects of p210 BCR/ABL and IL-3 on the pattern of tyrosine phosphorylation of cellular proteins in 32Dcl3 cells using one- and two-dimensional antiphosphotyrosine immunoblotting. WEHI-3B conditioned media (WEHI-CM) was used as a source of IL-3. The introduction of p210 BCR/ABL results in constitutively increased levels of tyrosine phosphorylation of more than 20 new proteins, while WEHI-CM induced transient tyrosine phosphorylation of 6 to 10 new proteins. Using two-dimensional immunoblots to examine phosphoproteins, four categories could be identified: (1) proteins that are inducibly tyrosine phosphorylated in response to WEHI-CM in 32Dcl3 cells only, (2) proteins inducibly tyrosine phosphorylated by WEHI-CM only in p210 BCR/ABL+ cells, (3) proteins that are inducibly tyrosine phosphorylated in response to WEHI-CM in both 32Dcl3 cells and p210 BCR/ABL+ cells, and (4) proteins inducibly tyrosine phosphorylated in response to WEHI-CM and constitutively phosphorylated in the presence of p210 BCR/ABL. We have identified one of the proteins in category 4 as p42 mitogen-activated protein (MAP) kinase (ERK2). Overall, however, we found that the signal transduction pathways of IL-3 and BCR/ABL are strikingly different, suggesting that most of the immediate substrates of the IL-3 receptor-activated tyrosine kinase and p210 BCR/ABL kinase are different. Convergence of signaling pathways at p42 MAP kinase is of interest since activation of this kinase has been linked to mitogenesis in many systems. Identification of the overlapping proteins of both IL-3 signal transduction in 32Dcl3 cells and p210 BCR/ABL+ cells may help explain the growth promoting effects of this oncogene. PMID- 8405227 TI - The effects of treatment with PIXY321 (GM-CSF/IL-3 fusion protein) on human polymorphonuclear leukocyte function. AB - During a phase I trial of the genetically engineered hematopoietic growth factor PIXY321 (granulocyte-macrophage colony-stimulating factor/interleukin-3 [IL-3] fusion protein), we examined the effects of PIXY321 treatment on human polymorphonuclear leukocyte (PMN) locomotive, respiratory burst, and phagocytic responses. PIXY321 treatment was associated with transient suppression of both unstimulated locomotion and chemotaxis responses to multiple stimuli, as well as significant transient enhancement of formyl peptide-stimulated H2O2 production. No effects on opsonic phagocytosis of Staphylococcus aureus were observed. In vitro exposure of control PMN to PIXY321 resulted in suppression of unstimulated locomotion/chemotaxis and enhancement of formyl peptide-stimulated H2O2 production but had no effects on phagocytosis. When patient cells were exposed in vitro to PIXY321 during treatment, suppression of chemotaxis and enhancement of H2O2 production were observed before PIXY321 treatment, but these effects diminished during treatment. The in vivo and in vitro exposure effects of PIXY321 treatment on PMN function are similar to those of the parent molecule, granulocyte-macrophage colony-stimulating factor (GM-CSF). PMID- 8405228 TI - Effects of recombinant human erythropoietin and interleukin-3 on erythropoietic recovery from acute anemia. AB - The risks inherent in the use of homologous blood products have increased efforts toward identifying alternatives to transfusion. We have previously shown that the administration of recombinant human erythropoietin (rhEpo) enhances the erythropoietic response to acute blood loss. Recombinant human interleukin-3 (rh IL-3) is a hematopoietic growth factor that has been shown to act synergistically with rhEpo in accelerating erythropoiesis in vitro. The purpose of this study in a primate model was to determine if the administration of rhIL-3 in combination with rhEpo could augment the erythropoietic response to acute blood loss more than rhEpo therapy alone. Twenty-four adult male baboons were randomized into four groups. The induction of acute normovolemic anemia to a hematocrit of 20% was accomplished via exchange-transfusion with 6% hetastarch. The groups were then treated for 7 consecutive days with the following growth factors: group I (n = 7), no growth factors; group II (n = 5), rhIL-3 alone (100 micrograms/kg/d); group III (n = 6), rhEpo alone (1000 U/kg/d); group IV (n = 6), rhEpo (1000 U/kg/d) plus rhIL-3 (100 micrograms/kg/d). All animals received folate, vitamin B12, and intravenous iron-dextran immediately following the exchange-transfusion. Response to therapy was monitored for 35 days. There were no adverse reactions following growth factor administration. The analysis of erythropoietic rates between study days 1 through 11, as determined via linear regression analysis, revealed that hematocrits increased significantly faster in the groups receiving rhEpo compared to controls. The administration of rhIL-3, however, did not increase the rate of erythropoiesis when compared to controls, nor did it augment response when added to the rhEpo regimen. The results of this study demonstrate that the administration of rhIL-3 alone had no significant effect on erythropoiesis in this setting of acute blood loss. Further, despite promising in vitro data, rhIL-3 provided no additional stimulation of erythropoiesis in animals receiving rhEpo. Nevertheless, the study confirms that the pharmacologic acceleration of erythropoiesis by rhEpo alone remains an attractive alternative to homologous transfusion. PMID- 8405229 TI - Fc gamma RIIA-mediated phagocytosis and receptor phosphorylation in cells deficient in the protein tyrosine kinase Src. AB - In the absence of other Fc receptors, stimulation of Fc gamma RIIA induces receptor phosphorylation and phagocytosis of immunoglobulin G (IgG)-coated cells. In vitro, Fc gamma RIIA is phosphorylated by the Src-related tyrosine kinase (SRTK) Src. Therefore, we investigated whether fibroblasts transfected with Fc gamma RIIA mediate phagocytosis of IgG-coated cells and whether Src is required for Fc gamma RIIA phosphorylation and for phagocytosis in vivo. Activation of Fc gamma RIIA in a fibroblast cell line deficient in Src kinase resulted in phosphorylation of the receptor on tyrosine. In addition, Fc gamma RIIA-mediated phagocytosis was observed in these fibroblasts in both the presence and absence of Src. In the presence of Src, however, phagocytosis of IgG-coated cells was more efficient. The data indicate that the SRTK Src is not required for Fc gamma RIIA phosphorylation or for Fc gamma RIIA-mediated phagocytosis in these cells. In vitro kinase assays demonstrated that the SRTK Fyn also is able to phosphorylate Fc gamma RIIA. Thus, Fc gamma RIIA can be phosphorylated by more than one tyrosine kinase in vitro. The data suggest that there may be shared functions among some intracellular kinases in receptor phosphorylation. PMID- 8405230 TI - Transfection of interferon-gamma gene in a mouse bone marrow stromal preadipocyte cell line causes apoptotic cell death. AB - It has been reported that bone marrow and serum of patients with aplastic anemia or chronic myeloproliferative disorders contain an abnormal concentration of cytokines. In the present study, we tried to isolate mouse bone marrow stromal cell lines that were stably transformed with a variety of cytokine genes and that expressed them constitutively. From mouse bone marrow stromal cell lines MBA-1, MBA-13, and 14F1.1, we isolated clones secreting interleukin-3 (IL-3), IL-4, granulocyte-macrophage colony-stimulating factor (GM-CSF), or granulocyte (G) CSF. Interferon-gamma (IFN-gamma)-producing stable transformants could not be established from 14F1.1 cells in spite of repeated transfection trials. At early stages of transfection, 14F1.1 cells did secrete IFN-gamma; however, exogenously added mouse IFN-gamma could not inhibit 14F1.1 cell growth. We discovered that chromosomal DNA isolated from 14F1.1 after transfection with the mouse IFN-gamma gene was fragmented. This is characteristic of cells undergoing apoptotic cell death. DNA fragmentation was also observed in 14F1.1 cells transfected with the human IFN-gamma gene. These results indicate that intracellular IFN-gamma induces apoptotic cell death of 14F1.1 stromal cells. PMID- 8405231 TI - Hematopoietic survival factors. PMID- 8405232 TI - Minor histocompatibility antigens. PMID- 8405233 TI - Colony growth in cultures from bone marrow and peripheral blood after curative treatment for leukemia and severe aplastic anemia. AB - The recovery of colony-forming cell numbers after curative treatment for leukemia and severe aplastic anemia (SAA) was studied. We examined 191 patients (85 acute myeloid leukemia [AML], 48 acute lymphocytic leukemia [ALL], 32 chronic myeloid leukemia [CML], 17 SAA, and nine myelodysplastic syndrome [MDS]) who were in hematologic remission 6 months to 13 years after either curative chemotherapy (n = 69) or allogeneic bone marrow transplantation (BMT) (n = 122) by culturing their precursor cells from bone marrow (BM) (n = 548) and peripheral blood (PB) (n = 529) in methylcellulose. Thirty-six BM donors and 25 PB donors served as controls. BM colony-forming cell numbers were abnormally low in all patients (p < 0.002) irrespective of underlying disorder and type of treatment (chemotherapy or irradiation). These numbers did not normalize with time--colony-forming cells were still strongly reduced up to 10 years after therapy, whether or not the patient had received an allogeneic bone marrow graft (p < 0.002). We also compared patients who remained in stable hematologic remission with those who later relapsed (6 months to 2 years after treatment). BM colony-forming cell numbers were significantly lower in patients who subsequently relapsed (p = 0.004). In contrast to BM cultures, we found normal colony-forming capacity by PB precursors in all patients. We conclude that (1) after chemotherapy or BMT, colony-forming cell numbers of BM in culture are permanently reduced; (2) this defect is probably due to a dysfunction of the BM environment rather than to a numerical reduction of the precursor cell pool; and (3) very low colony-forming capacity may be related to relapse. PMID- 8405234 TI - Effects of CAMPATH-1 antibodies on the functional activity of monocytes and polymorphonuclear neutrophils. AB - The rat antihuman lymphocyte monoclonal antibodies CAMPATH-1M and CAMPATH-1G (IgM kappa and IgG2b kappa, respectively) recognize cell surface antigens (CDW52) present on normal T and B lymphocytes as well as on monocytes, with weak expression on neutrophils. In previous studies, when peripheral blood mononuclear cells were treated with CAMPATH-1M and human complement, more than 99% of lymphocytes were killed. The present study indicates that both CAMPATH-1M and -1G bind to peripheral blood neutrophils and monocytes and do not affect the activity of the former cells but decrease the functional activity of monocytes. Decreased functional activity includes suppressed superoxide production by monocytes in response to phorbol myristate acetate (PMA), reduced activation of the cells as indicated by decreased ability to reduce 3-(4.5-dimethylthiazol-2-yl)-2.5 diphenyl-tetrazolium bromide (MTT), reduced tumoricidal activity, and reduced capability to kill Candida albicans. The ability of CAMPATH-1 to suppress the functional activity of monocytes in vitro suggests that in vivo administration of CAMPATH-1G may also affect the function of monocytes. PMID- 8405235 TI - The cloning and expression of the gene for ovine interleukin-3 (multi-CSF) and a comparison of the in vitro hematopoietic activity of ovine IL-3 with ovine GM-CSF and human M-CSF. AB - The ovine interleukin-3 (IL-3) gene has been cloned. It encodes a protein 146 amino acids (aa) in length and is situated approximately 10 kb from the gene encoding granulocyte-macrophage colony-stimulating factor (GM-CSF). A comparison of the ovine gene sequence with that of the human IL-3 gene reveals that the protein coding regions share between 49 and 59% identity, while the introns and other noncoding regions share between 63 and 78% identity. The gene promoters also share approximately 72% identity. Recombinant ovine IL-3 (rovIL-3) was expressed in Chinese hamster ovary (CHO) cells, and its biologic activity was compared to that of rovGM-CSF and recombinant human macrophage colony-stimulating factor (rhM-CSF) on ovine bone marrow cells. rovIL-3 predominantly stimulated the growth and development of mast cells and macrophages in liquid cultures and colonies of mixed cell phenotype, megakaryocytes, erythroid burst-forming units (BFU-E), and basophilic granular cells in soft agar cultures of bone marrow cells. In common with rovGM-CSF, IL-3 also stimulated eosinophil and macrophage colonies, which were increased in size and number of cells in cultures containing both cytokines. Maximum macrophage colony numbers were achieved with the combination of rovIL-3, rovGM-CSF, and rhM-CSF. rovGM-CSF stimulated neutrophil colony formation, whereas rovIL-3 did not. PMID- 8405236 TI - 5-fluorouracil and mast cell precursors in mice. AB - In the mouse hematopoietic system, 5-fluorouracil (5-FU) reversibly inhibits the generation of multilineage colonies containing granulocyte, erythroid, megakaryocyte, and macrophage lineage. To determine the effect of 5-FU on mastopoiesis in vitro, bone marrow cells were obtained from mice, cultured, and treated with 5-FU for 14 days in an interleukin-3 (IL-3)-enriched medium. A dose related inhibitory effect of 5-FU on mastopoiesis was found. When an inhibitory dose (1 microgram/mL) of 5-FU was supplemented to the cultures for only 2, 4, or 8 days and the cells were then recultured without the drug, we observed inhibition of mastopoiesis directly related to the time of exposure of the cells to 5-FU. To determine the effect of 5-FU on mastopoiesis in vivo, bone marrow cells from mice that had received a single intravenous (i.v.) 5-FU injection (150 mg/kg) were cultured. A virtually total absence of mast cells was noted at days 1 and 2 following 5-FU administration. A gradual reappearance of mast cells was later observed. Whether mice were injected with the drug once or with four once daily (100 mg/kg 5-FU) injections, a similar pattern of delay of mast cell appearance was observed. The findings suggest (1) an irreversible, nonadditive, toxic effect of 5-FU on mast cell precursors and (2) that most or all of the mast cell precursors are nonquiescent cells, continuously activated or cycling. In addition, the use of 5-FU may serve as a unique model system for controlling and studying mastopoiesis in normal mice, rather than the mutated mice currently studied. PMID- 8405237 TI - Diminished DNA synthesis in T cells from B chronic lymphocytic leukemia after phytohemagglutinin, anti-CD3, and phorbol myristate acetate mitogenic signals. AB - T lymphocytes from patients with B cell chronic lymphocytic leukemia (B-CLL) exhibit defective proliferative response to plant lectins. The blastogenic response of purified T lymphocytes to signals that interact with membrane molecules (phytohemagglutinin [PHA], anti-CD3 monoclonal antibody [MAB]) and with the intracytoplasmic protein kinase C (PKC) was investigated in 22 B-CLL patients and 18 healthy controls. 3H-thymidine uptake in T lymphocytes from 14 of 22 B-CLL patients after PHA, anti-CD3, and phorbol myristate acetate (PMA) was found to be lower than in the healthy controls. This defective proliferative response was not corrected by the exogenous addition of interleukin-2 (IL-2) to the culture medium. In analyzing the cell cycle progression of these T lymphocytes from B-CLL patients, we found that the percentage of cells in S phase at 2 days of PHA stimulation was significantly decreased and that it was normalized after 5 days of culture. Defective response of T lymphocytes from B-CLL patients to polyclonal mitogens was observed in those patients with advanced disease (stages A, B, and C). However, this T cell proliferative response was normal in patients with "smoldering B-CLL." We conclude that the defective proliferative response to membrane and intracytoplasmatic mitogenic signals on T lymphocytes from a part of B-CLL patients can be ascribed to delayed activation and cell-cycle progression, and an association between the alterations in the T cell compartment of B-CLL patients and the progression of the disease may be suggested. PMID- 8405238 TI - Effect of donor and recipient gender disparities on fatal graft-vs.-host disease in a mouse model for major histocompatibility complex-matched unrelated-donor bone marrow transplantation. AB - Sex chromosome-linked minor histocompatibility determinants have been shown to affect the incidence and severity of graft-vs.-host disease (GVHD) in both humans and animals. On the basis of earlier studies done in mice and humans, it has often been assumed that this effect is due to a simple response of female donor cells recognizing recipient male HY antigens as foreign and reacting against them. However, the data of various clinical groups have not always supported this assumption. Moreover, since most of the earlier mouse studies focused only on the single transplant direction of female into male and/or were done under totally syngeneic conditions, the possibility of a GVHD response based on donor recognition of the recipient female HX antigen as foreign was never fully addressed. We have therefore reexamined the question in a more clinically relevant allogeneic transplantation setting, using a major histocompatibility complex (MHC)-matched, unrelated-donor mouse model. Five different donor/recipient sets were paired in all four possible gender combinations. The results indicated that, in addition to GVHD reaction against male HY, reaction against female HX was also possible. The results also showed that when the total level of GVHD due to autosomal chromosome minor histocompatibility disparities is extensive, it may masks the influence of gender-related factors on GVHD. Finally, the data also suggest the possibility that the sex chromosome-linked minor histocompatibility determinants may be polymorphic and thus capable of multiple allele expression. PMID- 8405239 TI - Effects of recombinant human granulocyte-macrophage colony-stimulating factor on platelet survival and activation using a nonhuman primate model. AB - In humans and nonhuman primates, the in vivo administration of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) consistently results in marked increase of megakaryocyte ploidy and size similar to that observed with interleukin-6 (IL-6). However, whereas the administration of IL-6 also results in an increase in circulating platelets, there is no predictable corresponding increase in peripheral blood platelets following treatment with rhGM-CSF. To determine whether the failure of rhGM-CSF to produce thrombocytosis is secondary to cytokine-related increased platelet activation and consumption in vivo, we quantified autologous platelet survival time and in vivo platelet activation before and during 5 days of administration of rhGM-CSF to two rhesus monkeys. Platelet survival was measured using autologous platelets labeled with 111Indium oxine. Platelet activation was assessed by flow cytometric determination of the expression of the major platelet membrane glycoprotein (GP) IIb/IIIa complex, and an activation-dependent epitope on GPIIb/IIIa (recognized by monoclonal antibodies [MABs] LJ-P4 and PAC1, respectively). Platelet activation was also assessed by dose-response aggregometry using adenosine diphosphate (ADP). While megakaryocyte ploidy increased during rhGM-CSF administration, peripheral platelet counts were 418 x 10(9)/L and 525 x 10(9)/L before and 402 x 10(9)/L and 508 x 10(9)/L during cytokine treatment in animals 1 and 2, respectively. No changes were observed in the mean platelet volume. 111Indium-labeled platelet recovery in circulation was similar before (94.7%, 91.8%) and during (92.9%, 92.8%) rhGM-CSF administration, which indicates that cytokine-related in vivo sequestration of platelets does not occur. Autologous platelet survival was 5.6 and 6.2 days before and 5.0 and 5.4 days during the rhGM-CSF treatment (p = 0.07), without significant change in the corresponding platelet turnover rate (derived from the platelet count and survival time). The flow cytometric analysis showed no increase in the binding of either LJ-P4 or PAC1 MABs to the platelet membrane during rhGM-CSF administration. The aggregometry studies demonstrated similar concentrations of ADP inducing half-maximal aggregation (ED50). Overall, the above data indicate that treatment with rhGM-CSF is not associated with in vivo activation, sequestration, or increased consumption of platelets. The data suggest that the failure of rhGM-CSF-stimulated megakaryocytes to increase peripheral platelet count is a manifestation of ineffective megakaryocytopoiesis resulting from inability to increase platelet delivery to the circulation. PMID- 8405240 TI - Induction of murine peritoneal macrophage colony-forming cells by peritoneal administration of macrophage inflammatory protein-1 alpha. AB - Peritoneal injection of thioglycollate medium (TM) to mice results in a dramatic increase in total number of peritoneal macrophages within 48 to 72 hours. Unlike resident macrophages, a fraction (10 to 20%) of these newly arrived young macrophages, designated as macrophage colony-forming cells (M-CFC), are highly proliferative and formed macrophage colonies in vitro in the presence of either macrophage or granulocyte-macrophage colony-stimulating factor (M-CSF or GM-CSF). Using a reverse transcriptase polymerase chain reaction (RT-PCR) technique, peritoneal exudate macrophages (PEM) obtained 2 to 5 days after a single TM injection actively expressed mRNA for recombinant murine macrophage inflammatory protein-1 alpha (rmMIP-1 alpha). Yet none or only a trace amount of mRNA for MIP 1 alpha was detected in normal resident macrophages or PEM obtained 7 days after TM treatment. The effect of rmMIP-1 alpha on the induction of exudate M-CFC was investigated. Multiple intraperitoneal (IP) administration of rmMIP-1 alpha caused a marked increase in the total number of peritoneal M-CFC and macrophages similar to but weaker than the increase in TM-injected mice. The total number of neutrophils, mast cells, and eosinophils also increased, but with different kinetics, following multiple injections of rmMIP-1 alpha. rmMIP-1 alpha alone did not stimulate the proliferation of M-CFC, nor did it potentiate their responsiveness to either rmGM-CSF or recombinant human (rh) M-CSF in vitro. Taken together, our results suggest that MIP-1 alpha released by exudate macrophages is a major chemoattractant responsible for the migration of M-CFC from the circulation to the peritoneal cavity during a TM-induced inflammatory response. PMID- 8405241 TI - Macrophage colony-stimulating factor stimulates growth progression of the G1 phase fraction and induces monocytic differentiation of the G2/M-phase fraction in human myeloid leukemia cells. AB - The human myeloid leukemia cell line (NKM-1) proliferates in response to exogenous macrophage colony-stimulating factor (M-CSF) in serum-free medium. This CSF alone, however, does not induce any feature of cell differentiation in NKM-1 cells. We showed that terminal monocytic differentiation is partially observed only when NKM-1 cells are sequentially treated with 1,25-dihydroxyvitamin D3 and M-CSF, as evaluated by cell morphology, analysis of surface antigens, and phagocytic function. To determine whether there might be an association between the actions of M-CSF and special phases of the cell cycle, we separated D3 treated cells into cycle-specific fractions by centrifugal elutriation. G1 enriched cells were mostly stimulated for growth progression with M-CSF. On the other hand, G2/M-enriched cells were significantly induced into monocytic differentiation. The cell percentage in the G2/M phase has clearly correlated with that of terminally differentiated cells by M-CSF. These results suggest that two distinct actions of M-CSF to stimulate cell proliferation and differentiation could work on different phases of the cell cycle: cell growth in the G1 phase and cell differentiation in the G2/M phase. PMID- 8405242 TI - Local cerebral glucose utilisation following acute and chronic bilateral carotid artery ligation in Wistar rats: relation to changes in local cerebral blood flow. AB - The effects on local cerebral blood flow (LCBF) and glucose utilisation (LCGU) of permanent, bilateral carotid artery ligation (BCAL) were studied in conscious Wistar rats. LCBF and LCGU were measured using quantitative autoradiographic 14C iodoantipyrine and the 14C-2-deoxyglucose (14C-DG) techniques in 24 anatomically discrete regions of the brain. LCBF in the cerebral hemispheres 2.5 h (acute) after BCAL significantly decreased to 25-87% of the sham control, with the exception of the mammillary body. After acute BCAL, there was a heterogeneous accumulation of 14C-DG in the caudate nucleus and cerebral cortices. Only in the lateral geniculate body did LCGU significantly decrease after BCAL. One week (chronic) later, LCBF was significantly decreased in 15 (containing the caudate nucleus and all the cerebral cortices) of 24 structures. LCGU in ten (containing the caudate nucleus and all the cerebral cortices) of 24 structures after chronic BCAL significantly decreased to 66-77% of the sham control, except for regions with neuronal damage in which there was a heterogeneous uptake of 14C-DG. The ratio of LCBF/LCGU in chronic BCAL was unchanged in comparison with values in the corresponding sham-operated group. This model of acute and chronic cerebral ischaemia, with impairment in cerebral circulation and/or glucose metabolism, is expected to become a pertinent tool for the neurophysiologist. PMID- 8405243 TI - Error accumulation and error correction in sequential pointing movements. AB - Human subjects pointed, without seeing their arm, at visual targets presented in repeated sequences in a frontal plane. Required movement direction could change within the sequence by 0, 45, 90, 135 or 180 degrees. Hand position was recorded contact-free in three dimensions (3D). From the recordings, the pointing errors towards each target were transformed into a Cartesian coordinate system with the x-axis representing the mean direction of all movements towards that target. We then investigated the relationship between successive errors by applying linear regression analysis separately to the three Cartesian error components. For the x component, we found that successive errors were positively correlated throughout the experiment, which confirms our previous finding that errors in sequential pointing tend to accumulate (Bock and Eckmiller 1986; Bock et al. 1990). Correlation dropped by nearly 50% following a direction change of 90 degrees or more, suggesting that accumulation is reduced but not abolished by large changes in movement direction. The slope of the regression line averaged 0.6, which indicates the existence of a complementary trend towards error correction, contributing about 40% towards motor performance. Changes of movement direction affected slope and correlation in a closely similar way, suggesting that reduced accumulation is paralleled by increased correction. For the y- and z-components, we found that successive errors were positively correlated as well, but were not reduced following even large direction changes. This apparent discrepancy can be resolved by assuming separate neural mechanisms for amplitude and for direction control, differing in their sensitivity to direction changes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405244 TI - Motor cortical activity preceding a memorized movement trajectory with an orthogonal bend. AB - Two monkeys were trained to make an arm movement with an orthogonal bend, first up and then to the left ([symbol: see text]), following a waiting period. They held a two-dimensional manipulandum over a spot of light at the center of a planar working surface. When this light went off, the animals were required to hold the manipulandum there for 600-700 ms and then move the handle up and to the left to receive a liquid reward. There were no external signals concerning the "go" time or the trajectory of the movement. It was hypothesized that during that period signs of directional processing relating to the upcoming movement would be identified in the motor cortex. Following 20 trials of the memorized movement trajectory, 40 trials of visually triggered movements in radially arranged directions were performed. The activity of 137 single cells in the motor cortex was recorded extracellularly during performance of the task. It was found that 62.8% of the cells changed activity during the memorized waiting period. During the waiting period, the population vector (Georgopoulos et al. 1983, 1984) began to grow approximately 130 ms after the center light was turned off; it pointed first in the direction of the second part of the memorized movement (<--) and then rotated clockwise towards the direction of the initial part of the movement (increases). These findings indicate processing of directional information during the waiting period preceding the memorized movement. This conclusion was supported by the results of experiments in ten human subjects, who performed the same memorized movement ([symbol: see text]). In 10% of the trials a visual stimulus was shown in radially arranged directions in which the subjects had to move; this stimulus was shown at 0, 200, and 400 ms from the time the center light was turned off. We found that as the interval increased the reaction time shortened for the visual stimulus that was in the same direction as the upward component of the memorized movement. PMID- 8405245 TI - Modulation of grip force with load force during point-to-point arm movements. AB - In this paper, we examine grip forces and load forces during point-to-point arm movements with objects grasped with a precision grip. We demonstrate that grip force is finely modulated with load force. Variations in load force arise from inertial forces related to movement; grip force rises as the load force increases and falls as load force decreases. The same finding is observed in vertical and horizontal movements performed at various rates. In vertical movements, maximum grip force coincides in time with maximum load force. The maxima occur early in upward and later in downward movements. In horizontal movements, where peaks in load force are observed during both the acceleratory and deceleratory phases, grip force rises at the beginning of the movement and remains high until the end. The results suggest that when moving an object with the hand the programming of grip force is an integral part of the planning process. PMID- 8405246 TI - The effects of head and trunk position on torsional vestibular and optokinetic eye movements in humans. AB - We measured torsional vestibular and optokinetic eye movements in human subjects with the head and trunk erect, with the head supine and the trunk erect, and with the head and trunk supine, in order to quantify the effects of otolithic and proprioceptive modulation. During active head movements, the torsional vestibulo ocular reflex (VOR) had significantly higher gain with the head upright than with the head supine, indicating that dynamic otolithic inputs can supplement the semicircular canal-ocular reflex. During passive earth-vertical axis rotation, torsional VOR gain was similar with the head and trunk supine and with the head supine and the trunk erect. This finding implies that static proprioceptive information from the neck and trunk has little effect upon the torsional VOR. VOR gain with the head supine was not increased by active, self-generated head movement compared with passive, whole body rotation, indicating that the torsional VOR is not augmented by dynamic proprioceptive inputs or by an efference copy of a command for head movement. Viewing earth-fixed surroundings enhanced the torsional VOR, while fixating a chair-fixed target suppressed the VOR, especially at low frequencies. Torsional optokinetic nystagmus (OKN) evoked by a full-field stimulus had a mean slow-phase gain of 0.22 for 10 degrees/s drum rotation, but gain fell to 0.06 for 80 degrees/s stimuli. Despite this fall in gain, mean OKN slow-phase velocities increased with drum speed, reaching maxima of 2.5 degrees/s-8.0 degrees/s in our subjects. Optokinetic after nystagmus (OKAN) was typically absent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405247 TI - Primate frontal cortex: neuronal activity following attentional versus intentional cues. AB - We examined neuronal activity in three parts of the primate frontal cortex: the dorsal (PMd) and ventral (PMv) premotor cortex and a ventrolateral part of the dorsolateral prefrontal (PF) cortex. Two monkeys fixated a 0.2 degrees white square in the center of a video display while depressing a switch located between two touch pads. On each trial, a spatial-attentional/mnemonic (SAM) cue was presented first. The SAM cue consisted of one 2 degrees x 2 degrees square, usually red or green, and its location indicated where a conditional motor instruction would appear after a delay period. The stimulus event containing the motor instruction, termed the motor instructional/conditional (MIC) cue, could be of two general types. It might consist of a single 2 degrees x 2 degrees square stimulus identical to one of the SAM cues presented at the same location as the SAM cue on that trial. When the MIC cue was a single square, it instructed the monkey to move its forelimb to one of the two touch pads according to the following conditional rule: a green MIC cue meant that contact with the right touch pad would be rewarded on that trial and a red MIC cue instructed a movement to the left touch pad. Alternatively, the MIC cue might consist of two 2 degrees x 2 degrees squares, only one of which was at the SAM-cue location: in those cases, one square was red and the other was green. The colored square at the SAM cue location for that trial was the instructing stimulus, and the other part of the MIC cue was irrelevant. When, after a variable delay period, the MIC cue disappeared, the monkey had to touch the appropriate target within 1 s to receive a reward and could break visual fixation. The experimental design allowed comparison of frontal cortical activity when one stimulus, identical in retinocentric, craniocentric, and allocentric spatial location as well as all other stimulus parameters, had two different meanings for the animal's behavior. When a stimulus was the SAM cue, it led to either a reorientation of spatial attention to its location, or the storage of its location in spatial memory. By contrast, when it was the MIC cue, the same stimulus instructed a motor act to be executed after a delay period. For the majority of PMd neurons (55%), post-MIC cue activity exceeded post-SAM cue activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405248 TI - Hemispheric control of unilateral and bilateral responses to lateralized light stimuli after callosotomy and in callosal agenesis. AB - Normally, simple digital or manual responses to a light stimulus in the right or left visual hemifields are performed faster with uncrossed hand-field combinations than with crossed hand-field combinations. Because of the organization of visual and motor pathways, the integration of uncrossed responses is assumed to occur within a single hemisphere, whereas a time-consuming interhemispheric transfer via the corpus callosum is considered to be necessary for the integration of crossed responses. However, callosal transfer may be dispensable for those crossed responses which can be controlled through ipsilaterally descending motor pathways by the hemisphere receiving the visual stimulus. We investigated crossed-uncrossed differences (CUDs) in speed of simple visuomotor responses to lateralized flashes in one subject with total section of the corpus callosum and two subjects with complete callosal agenesis. We recorded the reaction times as well as the premotor times, as indicated by the electromyographic latencies of the prime movers, of three types of responses: a distal response involving a thumb flexion, a proximal response chiefly involving a forearm flexion and an axial response involving a shoulder elevation. Further, the three types of responses to a single lateralised flash were performed both unilaterally and bilaterally. The three acallosal subjects showed CUDs greatly exceeding normal values on distal responses, either unilateral or bilateral, and on unilateral proximal responses. These abnormally long CUDs stood in sharp contrast to the insignificant CUDs exhibited by the same subjects on bilateral proximal responses and on unilateral and bilateral axial responses in agreement with correspondingly insignificant CUDs reported for normal subjects. These results confirm that a callosal contribution is important for the execution of fast distal and unilateral proximal responses to a visual stimulus directed to the hemisphere ipsilateral to the responding hand. By contrast, the other types of crossed responses appear to be efficiently coordinated across the midline without the aid of the corpus callosum. This is in keeping with the hypothesis that they are governed by a bilaterally distributed motor system which is preferentially activated for the execution of symmetrical bilateral movements, employing axial and proximal limb muscles. PMID- 8405249 TI - Role of the different frontal lobe areas in the control of the horizontal component of memory-guided saccades in man. AB - Two paradigms of memory-guided saccades were studied in 14 patients with focal vascular lesions affecting either the frontal eye field (FEF), or the supplementary eye field (SEF) or Brodmann's area 46 in the prefrontal cortex (PFC), and in 13 age-matched control subjects. In the first paradigm, the subject had to remember the position of a visual target with the body immobile and, in the second, the position towards which gaze was directed before a body rotation, i.e. with a vestibular input. In control subjects, the percentage of error in saccade accuracy (horizontal component) was greater in the second than in the first paradigm (37% and 14% on average, respectively). Compared with controls, amplitude error was significantly increased in the FEF group for the first paradigm only, in the SEF group for the second paradigm only, and in the PFC group for both paradigms. These results are consistent with (1) the PFC providing an improvement in the utilization by the saccade system of the visual and vestibular signals used in the two paradigms, and (2) the FEF and SEF providing an improvement in the utilization of visual signals in the first paradigm and vestibular signals in the second paradigm, respectively. Furthermore, from these findings and experimental data, it may be hypothesized (1) that the PFC is a part of the network contributing to short-term memorization of both visual and vestibular signals, and (2) that the FEF and SEF control two different types of memory-guided saccades, with separate calculation modes to determine their amplitude. PMID- 8405250 TI - Within-arm somatotopy in human motor areas determined by positron emission tomography imaging of cerebral blood flow. AB - Within-arm somatotopy was identified in multiple motor areas of six normal human subjects who performed a visuomotor tracking task during positron emission tomography (PET) measurement of relative cerebral blood flow (relCBF). A randomly moving target, presented on a computer monitor, was continuously followed with the index finger (movement at the metacarpophalangeal joint), thumb, fist (movement at the wrist), forearm (movement at the elbow), elbow (movement at the shoulder), and eyes alone (control task) during sequential imaging. Segmental limb movements were associated with relCBF responses in the contralateral motor, supplementary motor, cingulate, and parietal cortex, and in the ipsilateral cerebellum. Localization of responses after stereotaxic transformation into Talairach atlas space, as well as within-subject analysis without anatomic deformations, demonstrated an overlapping somatotopic distribution in the motor cortex, with thumb responses most ventrolateral and shoulder responses most dorsomedial. Proximal limb movements induced relCBF responses of greater magnitude than distal movements. Somatotopy was also identified in the supplementary motor area, with index finger responses dorsal and anterior to shoulder responses. An additional set of somatotopic responses were located in the cingulate cortex, also with finger responses anterior to shoulder responses. Somatotopy was not identified in the anterior cerebellum. The distribution of relCBF responses is concordant with electrophysiologic studies in nonhuman primates that demonstrate a fractured somatotopy on a fine scale and a general somatotopic scheme of the limb on a large scale in multiple discrete motor areas. PMID- 8405251 TI - Single neurons are differently involved in stimulus-specific oscillations in cat visual cortex. AB - Synchronised oscillatory population events (35-80 Hz; 60-300 ms) can be induced in the visual cortex of cats by specific visual stimulation. The oscillatory events are most prominent in local slow wave field potentials (LFP) and multiple unit spikes (MUA). We investigated how and when single cortical neurons are involved in such oscillatory population events. Simultaneous recordings of single cell spikes, LFP and MUA were made with up to seven microelectrodes. Three states of single cell participation in oscillations were distinguished in spike triggered averages of LFP or MUA from the same electrode: (1) Rhythmic states were characterised by the presence of rhythmicity in single cell spike patterns (35-80 Hz). These rhythms were correlated with LFP and MUA oscillations. (2) Lock in states lacked rhythmic components in single cell spike patterns, while spikes were phase-coupled with LFP or MUA oscillations. (3) During non-participation states LFP or MUA oscillations were present, but single cell spike trains were neither rhythmic nor phase coupled to these oscillations. Stimulus manipulations (from "optimal" to "suboptimal" for the generation of oscillations) often led to systematic transitions between these states (from rhythmic to lock-in to non participation). Single cell spike coupling was generally associated with negative peaks in LFP oscillations, irrespective of the cortical separation of single cell and population signals (0-6 mm). Our results suggest that oscillatory cortical population activities are not only supported by local and distant neurons with rhythmic spike patterns, but also by those with irregular patterns in which some spikes occur phase-locked to oscillatory events. PMID- 8405252 TI - Primate frontal cortex: effects of stimulus and movement. AB - We compared neuronal activity in the dorsal premotor cortex (PMd), ventral premotor cortex (PMv), and prefrontal (PF) cortex of two rhesus monkeys. The behavioral design was a variant of the instructed delay task which established that: (1) a given visual stimulus could, on different trials, instruct different limb movements and (2) several different visual stimuli could instruct the same movement. Neurons in all frontal areas displayed the often replicated activity patterns that occur during instructed delay tasks, including phasic increases after instruction stimuli (signal-related activity), tonic discharge during an instructed delay period (set-related activity), and phasic premovement discharge (movement-related activity). For signal-, set-, and movement-related activity, the majority of neurons in PMd (51-64%), but only a minority in PF (16-18%) and PMv (32-40%), showed activity levels that significantly depended on the action instructed by that stimulus rather than simply the characteristics of the stimulus per se. Thus, most PMd activity, including the aspects that most resembled a sensory response, reflected factors in addition to the signal. Taken together with the results of related studies, it seems most likely that these other factors are dominated by the motor instructional significance of the stimulus. In addition, many neurons (17-37%) in all examined areas showed activity that significantly depended on which of various stimuli guided the same movement. This finding shows that, in those frontal areas, neuronal activity can be affected by both the action to be taken and the events guiding that action. PMID- 8405253 TI - Primate basal ganglia activity in a precued reaching task: preparation for movement. AB - Single cell activity was recorded from the primate putamen, caudate nucleus, and globus pallidus during a precued reaching movement task. Two monkeys were trained to touch one of several target knobs mounted in front of them after an LED was lighted on the correct target. A precue was presented prior to this target "go cue" by a randomly varied delay interval, giving the animals partial or complete advance information about the target for the movement task. The purpose of this design was to examine neuronal activity in the major structures of the basal ganglia during the preparation phase of limb movements when varying amounts of advance information were provided to the animals. The reaction times were shortest with complete precues, intermediate with partial precues, and longest with precues containing no information, demonstrating that the animals used precue information to prepare partly or completely for the reaching movement before the target go cue was given. Changes in activity were seen in the basal ganglia during the preparatory period in 30% of neurons in putamen, 31% in caudate nucleus, and 27% in globus pallidus. Preparatory changes were stronger and more closely linked to the time of movement initiation in putamen than in caudate nucleus. Although the amount of information contained in the precues had no significant effect on preparatory activity preceding the target go cue, a directional selectivity during this period was observed for a subset of neurons with preparatory changes (15% in putamen, 11% in caudate nucleus, 14% in globus pallidus) when the precue contained information about the upcoming direction of movement. A smaller subset of neurons showed selectivity for the preparation of movement amplitude. A larger number of preparatory changes showed selectivity for the direction or amplitude of movement following the target go cue than in the delay period before the cue. The intensity of preparatory changes in activity in many cases depended on the length of the delay interval preceding the target go cue. Even following the target go cue, the intensity of the preparatory changes in activity continued to be significantly influenced by the length of the preceding delay interval for 11% of changes in putamen, 8% in caudate nucleus, and 18% in globus pallidus. This finding suggests that preparatory activity in the basal ganglia takes part in a process termed motor readiness. Behaviorally, this process was seen as a shortening of reaction time regardless of precue information for trials in which the delay interval was long and the animals showed an increased readiness to move.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405254 TI - Angular velocity detection by head movements orthogonal to the plane of rotation. AB - Sinusoidal oscillation of rhesus monkeys about a head-fixed, earth-horizontal axis while rotating at constant velocity about an earth-vertical axis generates a characteristic ocular nystagmus where the three-dimensional slow phase eye velocity is compensatory to the spatially and temporally changing head angular velocity vector. This includes the generation of a unidirectional nystagmus characterised by a "bias" slow phase velocity component, albeit of small gain (0.2-0.7), that persists for the duration of the combined two-axes stimulation and is compensatory to the constant velocity earth-vertical axis rotation. Specifically, there is a torsional bias velocity in supine position, a vertical bias velocity in ear down position and a horizontal bias velocity in upright position. Since the semicircular canals can not sense prolonged constant velocity rotation, the ocular bias velocity must be centrally constructed from canal afferent signals using head position information. Thus, optimal performance of the vestibular system as a three-dimensional rate sensor relies on afferent information from both the semicircular canals and the otolith organs. PMID- 8405255 TI - Origin of the central entrainment of respiration by locomotion facilitated by MK 801 in the decerebrate rabbit. AB - In order to establish the origin of the central coupling between locomotion and respiration which operates in freely moving mammals during galloping, we sought experimental conditions that readily lead to such a coupling in decerebrate and curarised rabbit preparations. In such preparations, stimulation of the mesencephalic locomotor region (MLR) evokes locomotor activities, recorded from hindlimb muscle nerves, that are rarely totally coordinated with phrenic inspiratory activity. However, low doses (0.2 mg/kg i.v.) of MK 801, a non competitive NMDA antagonist which has been shown to increase the activity of the spinal locomotion generators (Fenaux et al. 1991), dramatically enhanced this coupling during MLR stimulation in most experiments: 1/1 coupling was dominant but 2/1 and 3/1 couplings (i.e. two or three locomotor cycles per respiratory cycle) were also obtained. Compared with spontaneous respiratory activity, which was apneustic under these conditions, the respiratory period was drastically decreased during coupling. However, a further transection of the spinal cord at the C6 or C7 level, which isolated the spinal locomotion generators from the supraspinal levels, totally suppressed this reduction of the inspiratory period during MLR stimulation in the presence of MK 801. In experiments where locomotor activity was simultaneously recorded at forelimb and hindlimb levels, the 1/1 evoked locomotor-respiratory coupling remained after the lumbar cord had been isolated by L1 spinal transection. The present data do show that intact spinal mechanisms are required for entrainment to occur. They suggest either tha a common supraspinal drive cannot entrain locomotion and respiration when being depressed, or that respiration is entrained at the locomotor rate by the spinal locomotion generators.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405256 TI - Postischaemic changes in protein synthesis in the rat brain: effects of hypothermia. AB - Protein synthesis, measured as [14C]-leucine incorporation into proteins, was studied in the normothermic rat brain following 15 min of transient cerebral ischaemia and 1 h, 24 h and 48 h of recirculation, and in the hypothermic (33 degrees C) brain following 1 h and 48 h of recirculation. Ischaemia was induced by bilateral common carotid occlusion combined with hypotension. Following normothermic ischaemia, incorporation of [14C]-leucine was depressed by 40-80% at 1 h of recirculation in all brain regions studied. At 48 h postischaemia, incorporation returned to normal or above normal levels in the inner layers of neocortex, the CA3 region, the striatum and the dentate gyrus, while in the outer layers of neocortex and in the hippocampal CA1 region the incorporation was persistently decreased by 26% and 40% respectively. At 24 and 48 h postischaemia, protein synthesis in the CA1 region and the striatum could be attributed to proliferating microglia. Intra-ischaemic hypothermia ameliorated the persistent depression of protein synthesis in the CA1 region at 48 h postischaemia, and a two-fold increase compared to the normothermic group was observed both in the CA1 region and the striatum. In the cortex, eucaryotic initiation factor 2 activity transiently decreased at 30 min postischaemia. In animals subjected to intra ischaemic hypothermia, the eucaryotic initiation factor 2 activity was reduced by 50% of control at 30 min of recirculation compared with 77% in normothermic animals. We conclude that the postischaemic depression of protein synthesis is in part caused by a decrease in eucaryotic initiation factor 2 activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405257 TI - Basic fibroblast growth factor enhances the growth of postnatal neostriatal GABAergic neurons in vitro. AB - Basic fibroblast growth factor (bFGF) significantly enhances the short-term survival of embryonic striatal neurons in vitro but has little effect on the outgrowth of striatal cells compared to neurons from other brain regions. Studies in our laboratory have shown that bFGF protects postnatal striatal cells in vitro from NMDA receptor-induced neurotoxicity. We therefore examined the effects of bFGF on the outgrowth of GABA-containing cells taken from the postnatal (Day 1) caudate-putamen and cultured for up to 3 weeks. In control cultures GABAergic neurons formed three populations based on somatic size and developed the cytoarchitectural features characteristic of dendrites, spines, and axons. In the presence of bFGF (6 pM continuously from the day of plating), small- and medium sized GABAergic neurons showed significant increases compared to untreated controls in axon-like growth (axon length) at 6 days in culture and in both axon- and dendrite-like neurite growth (axon length and branch order, number of primary dendrites, dendrite length, and dendritic branch order) at 13 and 17 days in culture. Large GABAergic neurons were unaffected by treatment with bFGF. Striatal GABAergic neurons exposed to nerve growth factor (10 ng/ml) were not different from untreated controls. Neuron survival was also unaffected by bFGF treatment at all days in culture examined. Other observations suggested that the neurotrophic effects of bFGF were mediated by a direct action of the growth factor on striatal neurons and not glial cells. First, glial cells (identified by the immunohistochemical localization of glial fibrillary acidic protein) were unaffected by bFGF treatment at the low concentration (6 pM) used to enhance neurite growth, but did significantly proliferate at higher concentrations of bFGF (6 nM). Second, immunoreactive bFGF receptor protein was localized predominantly to the somata and processes of striatal neurons and not to glial cells in the cultures. Finally, when neurons from control cultures were briefly exposed (1 to 4 h) to bFGF at concentrations which were neurotrophic, a marked elevation in the immediate early gene protein c-fos was observed by immunohistochemistry in the nuclei of neurons, including GABAergic cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405258 TI - Enhancement of c-fos expression in neurons of the rat spinal cord after partial denervation: evidence for functional plasticity. AB - c-fos immunocytochemistry was used to test a functional correlate of neuroplasticity involving nociceptive primary afferent fibers in the partially denervated adult rat spinal cord. Unilateral dorsal root ganglionectomies were made at L1-L4 (chronic side). After 3 weeks, contralateral L1-L4 ganglionectomies (acute side) were made 1 week prior to sacrifice. Two hours prior to perfusion, the animals were anesthetized and their hindlimbs were immersed in a 52 degrees C water bath for 20 s. The spinal cords were then processed for c-fos immunocytochemistry and the numbers of c-fos-immunoreactive cells determined. Following bilateral noxious thermal stimulation of the hindlimbs, the numbers of c-fos-immunoreactive cells in laminae I-II from L3 to L5 were increased by nearly twofold on the chronically deafferented side of the spinal cord (P < 0.05). This finding suggests augmented functional plasticity of nociceptive primary afferent fibers on the chronically denervated side of spinal cord. These observations are discussed with relation to recent demonstrations of enhanced calcitonin gene related peptide immunoreactivity and possible primary afferent fiber sprouting in similar denervation models. PMID- 8405259 TI - Reversibility of nerve growth factor's enhancement of choline acetyltransferase activity in cultured embryonic rat septum. AB - Choline acetyltransferase (ChAT) activity and survival of acetylcholinesterase (AChE)-positive neurons were measured in low-density cultures of embryonic (Day 14-15) rat septum exposed to various sequences of nerve growth factor (NGF) exposure and deprivation for up to 7 weeks in vitro. Most septal cultures grown 4 5 weeks with no exogenous NGF (including exposure to monoclonal or polyclonal antibodies against NGF) retained both a basal ChAT activity and the ability to increase ChAT activity in response to subsequently added NGF. When cultures were exposed to NGF (7S, 0.75 nM) for 2-3 weeks and then deprived of NGF for 2 weeks, ChAT activity fell gradually, but the number of AChE-positive neurons remained unchanged, and in many cases ChAT activity could be restored by subsequent re exposure to NGF. Thus NGF's enhancement of ChAT activity in embryonic septal neurons in vitro is largely reversible and is not mediated by differential survival of cholinergic neurons. PMID- 8405260 TI - Observations on the survival of grafted embryonic motoneurons in the spinal cord of developing rats. AB - The sciatic nerve of newborn rats was injured unilaterally. Small solid grafts of ED-12 embryonic spinal cord prelabeled with 5-bromo-2'-deoxyuridine (BrDUr) were inserted into the host's hemicord on the side of the sciatic nerve injury on PD5 12. Each graft was connected to a neuromuscular implant, which in group 1 consisted of the soleus muscle and its nerve taken from the healthy leg of the same rat pup, whereas in group 2 the soleus neuromuscular implant was taken from an immunocompatible adult rat. Six to 12 weeks later the neurons extending axons into the nerve-muscle implants were retrogradely labeled with fast blue and diamidino yellow. The embryonic origin of these neurons was ascertained by visualizing BrDUr using an immunocytochemical method. The grafts survived well and contained many BrDUr-positive neurons. No retrogradely labeled neurons were found in group 1 animals. In group 2 (animals with adult implants) numerous retrogradely labeled neurons were present in the host's neuropil. Despite the access to the target there were no BrDUr-positive cells of embryonic origin that contained the retrograde labels. Indeed no motoneuron-like cells of embryonic origin were seen in the host neuropil in any of the animals which received a graft at the age of 5-12 days. This contrasts with previous studies where the grafts were introduced into the adult spinal cord and motoneuron-like cells of embryonic origin were present in the host neuropil. Thus, it appears that the environment of the developing spinal cord is unfavorable for survival of embryonic motoneurons. PMID- 8405261 TI - The ability of developing spinal neurons to reinnervate a muscle through a peripheral nerve conduit is enhanced by cografted embryonic spinal cord. AB - The ability of neurons in the spinal cord of rats aged 5-12 days to reinnervate a muscle via a peripheral nerve bridge was examined and the possible influence of the cografted ED-12 embryonic spinal cord was tested. The soleus muscle was transferred paravertebrally and connected to the contralateral L4-L5 hemicord by its nerve. In some experiments embryonic spinal cord was grafted at the same level. Six to 12 weeks later fast blue and diamidino yellow were injected into the muscle or applied on the cut nerve bridge. The animals were perfused after 3 4 days and their spinal cords were examined using fluorescent microscopy, but retrogradely labeled neurons were only rarely seen. The embryonic spinal cord grafts survived well but had no influence on the outcome of these experiments. However, when neuromuscular implants from adult immunocompatible rats were used instead of the immature autologous ones, a variety of neurons including motoneurons extended their axons into the implants. The numbers of retrogradely labeled neurons were significantly higher in the spinal cords with embryonic grafts. These retrogradely labeled neurons were in the host's grey matter and only exceptionally in the grafts. Thus, the developing neurons can extend their axons outside the spinal cord into the implants of adult soleus muscle and nerve, but immature nerve-muscle implants fail to attract and/or support axonal outgrowth. The reinnervation potential of the host's spinal neurons was enhanced by cografting of embryonic spinal cord. PMID- 8405262 TI - Differential effects of MK-801 on brain-derived neurotrophic factor mRNA levels in different regions of the rat brain. AB - We have studied the effects of MK-801, a noncompetitive antagonist of N-methyl-D aspartate-type glutamate receptors, on brain-derived neurotrophic factor (BDNF) mRNA levels in the rat brain. MK-801 decreased BDNF mRNA in the hippocampus and in the superficial layers of the cerebral cortex. However, in single cells of the middle layer of the cerebral cortex and the midline thalamic nuclei BDNF mRNA levels were markedly increased by MK-801. The highest density of these cells was found in the limbic cortex, especially in the retrosplenial and medial entorhinal cortex. Pentobarbital (an enhancer of gabaergic functions) and scopolamine (a muscarinic receptor antagonist) blocked the effects of MK-801 on BDNF mRNA levels in the retrosplenial cortex, but the nicotinic and dopaminergic receptor antagonists mecamylamine and haloperidol, respectively, were ineffective. Pilocarpine, a muscarinic cholinergic agonist increased BDNF mRNA in some, but not all, cortical areas, where MK-801 had elicited an increase in BDNF mRNA. Thus, the observations made with MK-801 demonstrate that depending on the neuronal connections and the transmitter systems involved, a given compound can elicit either a decrease or an increase in BDNF mRNA levels. This may open up pharmacological possibilities to a regionally more refined regulation of the neurotrophin synthesis. PMID- 8405263 TI - Morphological and behavioral drawbacks of fetal dopaminergic grafts, prelabeled with Phaseolus vulgaris leucoagglutinin. AB - The anterograde tracer Phaseolus vulgaris Leucoagglutinin (Pha-L) was tested as a fetal cell marker in short-term labeling of a fetal dopaminergic cell suspension and in long-term surviving grafts in vivo. As a model we used the grafting of fetal dopaminergic cells into the denervated caudate putamen of the rat. Short term labeling revealed that the viability of the fetal cells was not affected by the Pha-L incubation within the 4 h of the test period. Yet, a subtle difference was noticed in the morphological development of the fetal neurons. Whereas many dopaminergic cells in the control suspension developed from an initially round soma to a more triangular or bipolar one, Pha-L-incubated cells maintained their round appearance. Moreover, cells with developing neurites were commonly noted in the control suspension, but were absent after incubation with Pha-L. Long-term effects of Pha-L were studied in three groups of unilaterally 6-hydroxydopamine lesioned rats, which all received an injection of a fetal dopaminergic cell suspension in the denervated caudate putamen. The first group (T-Pha-L) received dopaminergic cells, prelabeled with Pha-L. The second group (T-saline) received cells incubated with vehicle (saline). The third group (T) received only dissociated cells. Eight weeks after the implantation the morphological analysis showed a minor Pha-L-immunoreactivity inside the labeled grafts. We detected Pha L-positive fiber particles as well as weakly Pha-L-positive spots, presumably cell bodies. Pha-L-labeled grafts were significantly decreased in graft volume and contained markedly less dopamine-immunoreactive (DAi) cells than the control grafts of groups T-saline and T. The ratio DAi cell type I (cell with < or = 3 processes)/DAi cell type II (cell with > or = 4 processes) was approximately 8 in the control groups and 3 in group T-Pha-L. This suggests primarily a toxic effect of Pha-L and DAi cell type I neurons. Our behavioral data revealed that the Pha-L labeled grafts did not cause a recovery from lesion-induced motor asymmetries.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405264 TI - In vivo electrochemical measurements of exogenous dopamine clearance in normal and neonatal 6-hydroxydopamine-treated rat striatum. AB - The regulation of extracellular dopamine (DA) levels was studied in rat striatum after neonatal dopamine lesions and enhanced serotonin (5-HT) fiber ingrowth, induced with 6-hydroxydopamine (6-OHDA). We used rapid in vivo chronoamperometry combined with local DA applications. DA was pressure ejected in doses of 5 to 100 pmol at a distance of approximately 300 microns from the recording electrode, using single- or multibarrel glass micropipettes. Almost twice as much DA had to be applied in control rat dorsal and ventral striatum to obtain signals comparable to those recorded in the neonatal 6-OHDA-treated animals. In addition, in the dorsal striatum, the later portions of the DA clearance signals were significantly prolonged in the 6-OHDA group. Some clearance decay times in ventral striatum were also significantly prolonged in the neonatal 6-OHDA-treated rats. Concomitant application of the DA uptake inhibitor, nomifensine, in conjunction with the DA ejections, produced signal characteristics in the control striatum that were similar to those seen in the neonatal 6-OHDA-treated striatum. Taken together, these data support the hypothesis that a major component of the clearance of extracellular DA is dependent on intact terminals with high-affinity DA uptake, and that loss of DA afferents from neonatal 6-OHDA treatment results in a slowed clearance time of extracellular DA which is not apparently compensated by the enhanced 5-HT fiber ingrowth. PMID- 8405265 TI - Neurons derived from a human teratocarcinoma cell line establish molecular and structural polarity following transplantation into the rodent brain. AB - Studies of neurons grafted into the brains of experimental animals have been limited by the lack of suitably homogeneous populations of neurons for transplantation. Here we describe the transplantation and survival of pure, postmitotic human neurons (NT2N cells) into the rat brain. NT2N cells were derived from a human teratocarcinoma line (NTera2/clone D1 or NT2 cells) in vitro by retinoic acid treatment. Approximately 5-10 x 10(4) NT2N cells (including previously frozen aliquots of NT2N cells) were injected into the neocortex, subjacent white matter, or hippocampus of adult (N = 51) or neonatal (N = 17) Sprague-Dawley rats. Cyclosporine (7-10 mg/kg) was administered daily to 13 adult rats for up to 12 weeks post-transplant prior to sacrifice. Untreated rats survived for up to 21 weeks post-transplant. Injection sites were serially sectioned and NT2N grafts were analyzed immunohistochemically using antibodies to diverse neuronal and glial proteins to assess the lineage of the grafted cells and their ability to establish molecular and structural polarity. NT2N cells transplanted into untreated adult and neonatal rat brains were committed exclusively to the neuronal phenotype and survived for as long as 8 weeks, although most were rejected after 4 weeks. However, cyclosporine prolonged survival of the NT2N grafts for up to 12 weeks. Further, grafted NT2N cells exhibited an asymmetric geometry (with long axons and simplified dendrites), as well as molecular polarity (with highly phosphorylated neurofilament proteins segregated in axons and microtubule associated protein 2 confined to perikarya and dendrites) by 4 weeks post-transplant. However, the grafted neurons did not become fully mature as evidenced by their failure to express the most highly phosphorylated heavy neurofilament proteins. Finally, previously frozen NT2N cells survived in the rat brain, and none of the grafts formed neoplasms. We conclude from these studies that transplanted NT2N cells represent a highly advantageous model system for studies of the developmental biology of neurons. PMID- 8405266 TI - Survival of isolated axonal segments in culture: morphological, ultrastructural, and physiological analysis. AB - Some transected distal axons survive for months in vivo, generate new neurites, and reconnect to proximal segments before degenerating. To determine the factors regulating these phenomena, we studied the behavior of transected axons in cultured metacerebral neurons (MCn) of Aplysia. The neurons were isolated from the ganglia and cultured at 18 degrees C. The morphology, ultrastructure, and electrophysiological properties of the transected axons, as well as their ability to synthesize protein, were examined at different times postaxotomy. Follow-up studies revealed that cultured isolated axonal segments can preserve their morphological integrity for up to 14 days, maintain their passive and active membrane properties for at least 10 days, and extend new neurites and form electrotonic junctions with their proximal segments and intact MCns. De novo protein synthesis is an unlikely mechanism to account for the survival of the isolated axons since they did not incorporate [35S]methionine. We conclude that the viability of transected axons in culture devoid of other cells depends on pools of proteins synthesized prior to the transection and energy stores sufficiently large to maintain neuronal homeostasis. PMID- 8405267 TI - Neonatal undernutrition and amygdaloid nuclear complex development: an experimental study in the rat. AB - This study describes the morphology of neurons from the basolateral (ABL), central (ACE), and medial (AM) nuclei of the amygdaloid complex in neonatally undernourished (U) and control (C) Wistar strain rats. The cells were impregnated with the Golgi-Cox technique and studied at the ages of 12, 20, and 40 days postnatally. In the U-pups the neurons of the three nuclei displayed a reduced somatic area compared to that of the C-group on Days 12 and 20. However, at 40 days, this difference diminished due to a reduction in the somatic area of the C group. The dendritic area also appeared reduced on Days 12 and 20 in the U-group, but on Day 40 it reached control values. The neurons from ABL and ACE suffered a significant decrease in the number of dendritic branches due to undernutrition, but the AM nucleus did not show this change. The data suggest different vulnerability of these amygdaloid nuclei to neonatal undernutrition. The findings also suggest that the abnormal emotional response characteristic of perinatal undernourished rats could have a morphological cause. PMID- 8405268 TI - The cerebrovascular effects of physostigmine are not mediated through the substantia innominata. AB - This study sought to determine whether the cortical cholinergic projections from Meynert's nucleus are actually the target of the cholinesterase inhibitor physostigmine, which presents the ability to increase cortical blood flow. To this aim, the multiregional cerebrovascular effects of physostigmine in rats with and without lesion of the substantia innominata (SI), the equivalent of Meynert's nucleus of primates, were investigated. Unilateral SI lesions were made using ibotenic acid in three groups of rats. Four to 11 days later, the cortical choline acetyltransferase (ChAT) activity was measured in one group to assess the efficacy of the lesion. In the two other groups, the regional cerebral blood flow was measured using the [14C]iodoantipyrine technique, under physostigmine (0.2 mg/kg/h iv) or control conditions. SI lesion induced 27-59% fall in cortical ChAT activity in the ipsilateral hemisphere with the frontal area most affected. Despite these large biochemical differences, the lesion had little cerebrovascular effects. Side-to-side blood flow differences did not exceed 11% and did not strictly overlap the ChAT depletion. Physostigmine increased flow (38 66%) in all cortical areas, with no frontal predominance. Despite these considerable vasodilations, there were no significant differences between the lesioned and the intact hemisphere, nor any significant interaction between physostigmine and SI lesion. Thus, physostigmine does not actually activate the SI neuron terminals. This result suggests that cholinesterase inhibitors cannot be used as presynaptic markers of the cholinergic activity of this nucleus and casts doubts on their specificity as enhancement therapeutic agents in Alzheimer's disease. PMID- 8405269 TI - Avid binding of beta A amyloid peptide to its own precursor. AB - The senile plaque and congophilic angiopathy in brains of patients with Alzheimer's disease contain abnormal extracellular depositions. These depositions have very complex molecular structure, some elements of which have been identified. The most abundant protein in these structures is beta A amyloid peptide, which is produced by proteolytic processing of a larger protein, the amyloid precursor protein. It is not known how plaques and angiopathy are formed. Binding of proteins with the beta A peptide may be critical in this process. Using synthetic beta A peptides we show that the amyloid precursor protein found in cerebrospinal fluid binds beta A peptide with high avidity. Analysis of recombinant amyloid precursor proteins containing deletions of various domains demonstrate that the beta A peptide binds to the amino terminus of its own precursor. A model of plaque assembly is presented. PMID- 8405270 TI - Physiological studies in deafferented visual cortex cells of cats following transplantation of fetal xenografts from the rat's cortex. AB - We have studied the physiological properties of cells (N = 822) in visual cortex area 17 of seven adult cats transplanted with visual cortex xenografts from fetal (E15-E17) rats. The transplants were assumed to induce recovery of adjacent neurons partially deafferented from visual input. The control group (eight cats, 564 cells) had just analogous sectioning in the cortex. The level of activity found, following visual stimulation, in the deafferented cortical region (medially to the graft) was 48.6% compared to the result (34.5%) obtained in the control cats, indicating the preservation of visual responsiveness. Furthermore, no disturbance could be found in the ocular dominance distribution or binocularity (64.4%) of the cells in the grafted region compared to those in the control cats, indicating preservation of the columnar organization. The deafferented cells in the grafted cortex thus demonstrated the absence of adverse immunological reaction there due to the presence of the xenogeneic tissue, indicating that the visual cortex is immunologically privileged. PMID- 8405271 TI - Extension and regeneration of corticospinal axons after early spinal injury and the maintenance of corticospinal topography. AB - It has been established that neonatal corticospinal (CS) axons are able to grow around lesions of the spinal cord early in neonacy (Bernstein and Stelzner, J. Comp. Neurol. 221:382-400; Firkins, Bates, and Stelzner, Exp. Neurol., 120:1-15). To determine if these corticospinal axons include regenerating as well as late developing axons a double-labeling paradigm is used in which CS neurons are retrogradely labeled from the cervical spinal cord by injections of fast blue (FB) on Postnatal Day (PND) 2, 4, or 10. Two days later, the FB is aspirated along with the left dorsal funiculus and the right hemicord (CHR). As adults, the animals receive an injection of diamidino yellow (DY) or rhodamine into the spinal cord caudal to the lesion site. Thus, FB neurons are those that originally projected to the spinal cord before the lesion and which survived axotomy, DY neurons are those whose axons reached the spinal cord after the lesion, and double-labeled neurons (FB/DY) are cells which projected to the spinal cord prior to the lesion and regenerated a spinal axon postlesion. Animals FB injected on PND 2 have a widespread distribution of FB-labeled neurons in cortex, including areas outside of sensorimotor cortex. These animals also had both DY- and FB/DY labeled cells within sensorimotor cortex, indicating that the population of axons growing caudal to neonatal spinal lesions consists of both late growing and regenerating axons. In animals FB injected on PND 10, the FB neurons were all located in sensorimotor cortex. Very few DY and no FB/DY neurons were present. We have also looked at the topography of the CS neurons which project caudal to early spinal lesions. Rat pups received a CHR on PND 0, 3, 6, or 12. As adults, horseradish peroxidase was injected into the cervical or lumbar enlargement of the spinal cord and the distribution of labeled cells in the cerebral cortex was plotted and compared to normal and lesioned adult controls. In all experimental animals, the distribution of retrogradely labeled cells was restricted to the area containing CS projection neurons in the normal animal. This is despite the fact that the number of CS projection neurons is greatly reduced from normal and the normal pathway for CS axonal outgrowth has been completely disrupted by the neonatal lesion. PMID- 8405272 TI - Spinal cord transplants support the regeneration of axotomized neurons after spinal cord lesions at birth: a quantitative double-labeling study. AB - After spinal cord lesions in newborn rats, transplants of fetal spinal cord tissue rescue immature axotomized neurons, support the growth of axons into and through the site of injury and prolong the critical period for developmental plasticity. Both late-developing (uninjured) and regenerating axons contribute to this transplant-induced anatomical plasticity. After lesions in the mature CNS, transplant-induced axonal elongation is spatially restricted. The current study was designed (1) to determine the magnitude of transplant-induced regeneration, (2) to test the hypothesis that the long distance growth beyond the site of injury is mediated by late-developing axonal pathways, whereas axonal elongation by regenerating pathways is spatially restricted as it is in the adult, and (3) to determine if particular nuclei have a greater inherent capacity for regeneration than others. We used temporally spaced retrograde tracing with the fluorescent dyes fast blue and diamidino yellow to address this issue. Fast blue was placed into the site of a spinal cord overhemisection in rat pups < 48 h old to label those neurons which were axotomized by a neonatal lesion. The tracer was removed and a transplant of Embryonic Day 14 fetal spinal cord tissue was placed into the lesion site. Three to six weeks later a second tracer (diamidino yellow) was injected bilaterally into the host spinal cord caudal to the transplant. We counted the number of double-labeled (regenerated), single diamidino yellow labeled (late-growing), and single fast blue-labeled (nonbridging) neurons in the cortex, red nucleus, raphe nuclei, and locus coeruleus. By systematically varying the distance of the diamidino yellow injection site caudal to the transplant, we were able to compare the distance which injured axons regenerate with the distance that late-growing axons extend. When the diamidino yellow injection was placed within 5 mm caudal to the transplant 28% of the axotomized neurons in the red nucleus, 32% of the axotomized neurons in the locus coeruleus, and 37% of the axotomized neurons in the raphe nuclei were double-labeled (regenerating). Although the percentage of double-labeled neurons decreased as the distance beyond the transplant increased, a substantial population of regenerating neurons was identified in each of the brain stem nuclei examined following diamidino yellow injections up to 15 mm caudal to the transplant. Thus, after spinal cord lesions and transplants at birth, both regenerating neurons and late-developing neurons extended axons long distances (up to 15 m) caudal to the lesion site. The capacity for regenerative growth was similar in each of the nuclei examined. PMID- 8405273 TI - Time course of dorsal root axon regeneration into transplants of fetal spinal cord: an electron microscopic study. AB - Intraspinal transplants of fetal CNS tissue permit or enhance the regeneration of cut central axons of adult dorsal root ganglion (DRG) neurons. Some of these regenerated axons establish synapses with transplant neurons. The aims of the present study were to determine when regenerated DRG axons begin to form synapses with transplanted embryonic spinal cord neurons and whether these synapses are permanent. We also examined the development of transplant neuropil in areas innervated by the regenerated axons. Whole pieces of Embryonic Day 14 spinal cord were introduced into hemisection cavities made at the level of the lumbar enlargement, and the cut L4 or L5 dorsal root was juxtaposed to the transplant. Regenerated DRG axons immunoreactive for calcitonin gene-related peptide (CGRP) were labeled by immunohistochemical methods and examined by electron microscopy from 1 week to 1 year after surgery. CGRP-immunoreactive axon terminals made synaptic contacts with dendrites and perikarya of transplant neurons by 1 week after axotomy. The morphology of the synapses was immature. Large growth cone like structures were also present at 1 week but not at 2 weeks or later. At 2 weeks, regenerated unmyelinated axons formed terminals similar to those found in animals surviving for 48 weeks. Axoaxonic synapses in which the pre- and postsynaptic elements were immunolabeled for CGRP and regenerated CGRP-labeled myelinated axons were observed at 4 weeks and later. The area of distribution of CGRP staining increased until 12 weeks and the synaptic density of regenerated CGRP-labeled terminals increased for 24 weeks. The results indicate that the synaptic terminals of regenerated primary afferent axons are permanently retained within fetal spinal cord transplants. Transplants may therefore contribute to the permanent restoration of interrupted neural circuits. PMID- 8405274 TI - Adult spinal motoneurons remain viable despite prolonged absence of functional synaptic contact with muscle. AB - Several rat medial gastrocnemius (MG) motor axons were allowed to regenerate into normally innervated muscle. Under these conditions, synapse formation is known to be prevented by the existence of the original innervation of the host muscle. A study was made of the ability of the implanted spinal motoneurons to acquire and retrogradely transport horseradish peroxidase (HRP) injected into the host muscle at various postoperative intervals. HRP-labeled MG motoneurons on the implanted side were observed at postoperative intervals as long as 290 days. A comparison of the number of labeled MG motoneurons on the implanted side versus the number on the unoperated, control side indicated no significant differences. At all investigated postoperative intervals except the earliest (7 DPO), a significant decrease in the mean MG motoneuron soma cross-sectional area was observed relative to the unoperated, control side. Analysis of labeled motoneuron size distributions showed that postoperative atrophy of larger, presumably alpha, motoneurons occurred at a significantly faster rate than in smaller, presumably gamma, motoneurons. These results demonstrate that axotomized adult spinal motoneurons survive and remain viable for prolonged periods when denied the opportunity to reinnervate muscle but do so in an atrophied state. The results indicate further that alpha and gamma motoneurons differ quantitatively in their responses to peripheral axotomy. PMID- 8405275 TI - Calcitonin gene-related peptide increases following axotomy of trochlear motoneurons. AB - Neuropeptides have long been considered to act as neurotransmitters or neuromodulators, but they may also contribute to a variety of regulatory and trophic neuronal functions. In the present study, we determined the effects of axotomy on the levels of the neuropeptide calcitonin gene-related peptide (CGRP) in trochlear motoneurons (TMNs) of adult cats. The number of neurons with detectable CGRP immunoreactivity, and the intensity of their CGRP immunoreactivity, increased dramatically 1 week postaxotomy, gradually returned toward normal levels, but remained significantly higher than normal 12 weeks postaxotomy--a time when axonal regeneration and functional reinnervation of the superior oblique muscle should be complete. Our observation that CGRP levels in TMNs increase after axotomy suggests a role for this peptide in the response of motoneurons to injury and in regeneration. In addition, since many axotomized TMNs die, we suggest that the maintenance of high CGRP levels even after regeneration is complete may reflect an increased load on those TMNs that survive. PMID- 8405276 TI - The recovery of postural reflexes and locomotion following low thoracic hemisection in adult cats involves compensation by undamaged primary afferent pathways. AB - Spinal hemisection in the adult cat results in motor impairments followed by substantial recovery of function (16, 20, 39, 53). The present study was undertaken to assess the contribution of undamaged ipsilateral segmental and contralateral descending systems to recovery of motor function. Quantitative behavioral methods were used to examine monopedal reflex and bipedal locomotor functions after thoracic hemisection. Different facets of motor behavior recover at different times. The recovery of monopedal postural reflexes precedes the recovery of more complex motor behavior. Since the reflexes tested are initiated by segmental afferent input and show recovery and normal motor patterns during locomotion, as defined by kinematic analysis show recovery, it is likely that dorsal root input compensates for the loss of descending input to one side of the spinal cord. Quantitative immunocytochemical methods for visualizing the central projections of dorsal root fibers (monoclonal antibody RAT-102; 49) and the descending serotoninergic pathway were used to examine the response of these pathways to hemisection. Hemisection results in a permanent decrease in the density of serotoninergic projections and a permanent increase in dorsal root projections in the spinal cord. The increased density of RAT-102 may represent an increase in the projection of dorsal root fibers and provide the increased input necessary to mediate enhanced reflex control. A transient increase in GAP-43 in the dorsal horn ipsilateral to the hemisection suggests that the increased density of RAT-102 immunoreactivity is associated with growth. Taken together, our results suggest that sprouting of primary afferents within the spinal cord is one mechanism underlying the recovery of function after hemisection. PMID- 8405277 TI - Recovery of function after spinal cord injury: mechanisms underlying transplant mediated recovery of function differ after spinal cord injury in newborn and adult rats. AB - Fetal spinal cord transplants placed into the site of spinal cord injury support axonal growth of host systems in both newborn and adult animals. The amount of axonal growth, however, is much more robust in the newborn animals. The current studies were designed to determine if the differences in the magnitude of the anatomical plasticity of host pathways in the presence of transplants is reflected in differences in recovery of function between the neonatal and adult operates. Newborn and adult rats received a midthoracic "overhemisection." Immediately following the hemisection embryonic (E14) spinal cord transplants were placed into the lesion site. All animals were trained and tested as adults, on a battery of qualitative and quantitative tests of motor function. Immunocytochemical methods were used to compare the extent of growth of descending (serotonergic and noradrenergic) and segmental (calcitonin gene related peptide containing dorsal root axons) pathways in both groups. The growth of descending pathways into the transplants was substantially greater in density and spatial extent after lesions at birth than at maturity. The distribution of segmental dorsal root axons, in contrast, was similar in both groups. Fetal spinal cord transplants promoted recovery of motor function in both newborn and adult operates. The particular aspects of locomotor function which recover differ between the neonatal and adult operates, suggesting that the mechanisms underlying recovery of function must differ between the two groups. PMID- 8405278 TI - Evidence for alterations of synaptic inputs to sacral spinal reflex circuits after spinal cord transection in the cat. AB - Quantitative electron microscopy was used to study potential alterations in the synaptic inputs to HRP-labeled preganglionic neurons (PGNs) in the sacral parasympathetic nucleus (SPN) and to motoneurons (MNs) in Onuf's nucleus (ON) after short (4 days)- or longterm (10-11 weeks) spinal transection as compared to normal controls. Transection resulted in an apparent reorganization and replacement of synaptic input to ON MNs whereas chronic denervation of PGNs in the SPN was observed. These synaptic alterations may play a role in the changes in the eliminative reflexes (e.g., bladder-sphincter dyssynergia, induction of cutaneously elicited elimination) that are observed after spinal cord lesions. PMID- 8405279 TI - Reduced retrograde labeling of diencephalic-projecting neurons in the gracile nucleus of the monkey following removal of dorsal column input. AB - As part of an anatomical investigation of neuronal responses to deafferentation of the dorsal column nuclei by transection of the dorsal spinal columns, the uptake and retrograde transport of HRP by thalamic projection cells in the dorsal column nuclei was studied. The ventrobasal thalamus of 13 macaque monkeys was injected bilaterally with HRP at periods ranging from 3 to 364 days following intended unilateral transection of fasciculus gracilis at a mid- to upper thoracic level. The density of labeled cells in the gracile nuclei ipsilateral to complete lesions of fasciculus gracilis was compared with the density of labeled cells in the contralateral gracile nuclei that were fully innervated or partially denervated by an incomplete lesion. Also, the density of labeled cells in the fully innervated cuneate nuclei was compared. In general, there was a reduction in density of labeled cells in the gracile nuclei ipsilateral to complete lesions, without a corresponding decrement in labeled cells in the cuneate nuclei on that side. This result confirms effects on spinal motoneurons and on thalamocortical projection cells in the lateral geniculate nucleus following deafferentation. However, attempts to define a time course for the reduction in transport by lemniscal projection cells revealed an effect that was dramatic in some animals, partial in others, and not demonstrable in the remainder, without a clear relationship to time after surgery. This result is related to a literature which describes a variety of morphological, biochemical, electrophysiological, and behavioral effects of deafferentation which appear to wax and wane with time after neuronal injury. PMID- 8405280 TI - Partial deafferentation of cat spinal neurons results in permanent changes in cell surface molecular expression and metabolic activity. AB - Partial denervation of spinal neurons often results in reactive reinnervation by spared systems and some recovery of function. Dorsal rhizotomy in the adult cat has been used for the examination of the molecular modifications that occur postsynaptically and which may underlie anatomical and behavioral plasticity. We examined two markers of postsynaptic neurons in the spinal cord. Immunolocalization of a specific marker of superficial dorsal horn and intermediolateral neurons, the limbic system-associated membrane protein (LAMP), revealed that removal of certain afferents results in a permanent decrease in LAMP expression. Cytochrome oxidase, a metabolic marker, is normally high in the neurons of Clarke's nucleus, but showed a dramatic decrease after deafferentation. Reactive reinnervation occurs in many regions of the spinal cord following partial deafferentation, including Clarke's nucleus and the dorsal horn, and these changes in presynaptic input may be manifest by permanent modifications in molecular expression and metabolic activity of spinal neurons. PMID- 8405281 TI - Removal of dorsal root afferents prevents retrograde death of axotomized Clarke's nucleus neurons in the cat. AB - We investigated the effect of axotomy, deafferentation, and deafferentation plus axotomy on cell survival and cell size in Clarke's nucleus of the cat spinal cord. Hemisection of the adult spinal cord at T9 leads to retrograde cell death of 40% of the neurons in Clarke's nucleus at L3, as well as to a reduction in the mean soma size of the survivors. In contrast, deafferentation of Clarke's nucleus neurons by L1-S2 dorsal rhizotomy produces no cell loss and no shrinkage of the somata. These results indicate that dorsal root afferent input is not required for Clarke's nucleus cell survival. To test whether afferents may be required by the 60% of neurons that survive axotomy, we deafferented Clarke's nucleus prior to axotomy. Surprisingly, removal of primary afferents to Clarke's nucleus neurons prior to axotomy prevented the death of all neurons that would normally have died from axotomy. These results suggest that dorsal root afferent input is not required for Clarke's nucleus neuron survival after axotomy and may in fact be toxic to these axotomized neurons. This afferent toxicity is likely to be mediated through the dorsal root afferent neurotransmitter glutamate. PMID- 8405282 TI - Role of astroglial extracellular matrix in the formation of rat olfactory bulb glomeruli. AB - We describe the development of the olfactory bulb (OB) glomeruli, focusing on the role of purported astroglial-associated boundary molecules, as potential guidance factors in glomeruli formation. Glomeruli are unusual structures within the OB formed by extremely tight bundles of afferent fibers that fail to grow deeply into the brain. Using olfactory marker protein antibodies, we found that olfactory axons of stage E17-E18 rat embryos were organized into long strands at the outer-most edge of the bulb. By E20-E21 the axons clustered and formed small spheres and by PD1 they gathered into distinct glomeruli. Prior to and during the transformation of olfactory afferents into glomeruli, there was an enhanced expression of cytotactin/tenascin (CT) and chondroitin sulfate-containing proteoglycan (CS-PG) associated with VIM/GFAP-positive astroglial cell processes at the base of the glomeruli. During later stages of development (PD3-PD9), the GFAP-positive astroglial processes beneath and within the base of the glomeruli showed intensified GFAP staining while maintaining expression of CT and CS-PG. Some astroglial processes had extended and others went even further peripherally into the olfactory nerve layer (ONL) to encapsulate and fully infiltrate the glomeruli. By PD9, CS-PG and CT levels increased inside and outside the glomeruli. However, by PD15 both molecules had decreased inside the glomeruli but remained highly concentrated around the glomeruli to form rings. Thus, at later stages, axon/glia interactions which result in astroglial rearrangement and matrix alterations in relation to the glomeruli occur. Western blots of stage E20 bulb proteins revealed the presence of three proteoglycan core proteins each containing chondroitin-6-sulfate moieties. In addition, a type of keratan sulfate (KS) glycosaminoglycan was present in a core protein not associated with chondroitin-6-sulfate. Our data suggest that astroglial-associated ECM molecules CT and CS-PG, and perhaps KS-PG, present in the core of the early developing OB, could form a molecular "wall" that helps confine ingrowing olfactory axons within the ONL at the outer edge of the astroglial territory. We suggest that the astroglia do not precisely regulate the specific shape of the glomeruli, but rather play a fundamental role in directing the gross position at which formation of the synaptic glomeruli will occur. PMID- 8405283 TI - Degeneration of long-term fetal neostriatal allografts in the rhesus monkey: an electron microscopic study. AB - Primate fetal neostriatal neurons were implanted into the ibotenic acid-lesioned primate striatum and the animals were allowed to survive for 8 months. Light microscopic examination of the transplanted tissue demonstrated that the grafts were between 1.0 and 1.5 mm in diameter. The transplants were highly gliotic, but contained both normal appearing and degenerating neurons. At the electron microscopic level, the transplanted neurons displayed ultrastructural features identical to those of medium spiny, medium aspiny, and large aspiny striatal neurons. However, the majority of the grafted neurons showed evidence of degeneration. The grafts' neuropil demonstrated numerous glial processes, as well as mature axodendritic and axospinous synapses. Although this study was limited to only two graft recipients, the degenerative changes seen in the long-term primate allografts suggest that extension of these techniques into the clinical setting may be premature at the present time. PMID- 8405284 TI - A laminin graft replaces neurorrhaphy in the restorative surgery of the rat sciatic nerve. AB - We investigated the role of laminin in functional recovery of a peripheral nerve injury using electrophysiological and behavioral approaches on the rat sciatic nerve in vivo. These studies were complemented by neurofilament protein immunocytochemistry on the sciatic nerve 20 days after an operation, in which an 8-mm piece of the nerve was removed and replaced by a graft of laminin, its neurite outgrowth-promoting peptide, a control peptide, collagen, or by resuturing of the removed piece of the nerve. Electrophysiological measurements of muscle strength 4 months after the sciatic nerve transection showed that a laminin graft was as effective as neurorrhaphy in supporting functional recovery of an injured peripheral nerve. A laminin graft also significantly reduced autotomy in the operated animals. Immunocytochemistry confirmed that both a laminin graft and resuturing supported growth of the 200-kDa neurofilament positive axons into the distal stump of the nerve within 20 days of operation. A graft with a neurite outgrowth-promoting peptide of the B2 chain of laminin supported similar axon growth, whereas another peptide graft also derived from laminin or a collagen graft did not support axon growth. All grafts allowed Schwann cell growth into the distal stumps of the nerves, but neurites accompanied them only in the regeneration-supporting grafts and in the resutured nerves. The Schwann cells of the regenerating nerves expressed high levels of the neurite outgrowth-promoting domain of the B2 chain of laminin, whereas the Schwann cells of the degenerating nerves failed to express this domain in the distal stumps of the degenerating nerves. These results provide the first in vivo evidence for the functional role of laminin in peripheral nerve regeneration. As the neurite outgrowth-promoting domain of the B2 chain of laminin is as efficient as laminin or resuturing in supporting a short-term recovery of an injured sciatic nerve, this area may be a regeneration-promoting domain of this glycoprotein. More importantly, as grafting significantly reduces post-traumatic pain behavior in the operated animals, the laminin graft surgery may provide a useful method for clinical restoration of the injured peripheral nerves. PMID- 8405285 TI - Dopaminergic neuronal sprouting and behavioral recovery in hemi-parkinsonian rats after implantation of amnion cells. AB - Cells obtained from human, monkey, or rat term amnion membrane produce an activity which, in vitro, increases process outgrowth from rat sympathetic neurons and from dopaminergic neurons of the rat ventral mesencephalon. To determine if these cells could induce sprouting of dopaminergic nerve fibers in vivo, the substantia nigra of rats was lesioned unilaterally with 6 hydroxydopamine and live-rat-term amnion cells, or killed-rat-term amnion cells were implanted into the denervated striata. A control group of rats received saline injections into the denervated striata. Rats implanted with live amnion cells had a significant decrease in turning in response to amphetamine. The lesioned and implanted striata of live-amnion-cell-implanted rats contained significantly greater areas of tyrosine hydroxylase-immunoreactive fibers than the lesioned and implanted striatum of rats in the killed-amnion-cell or saline groups. Differences in the area of tyrosine hydroxylase-immunoreactive fibers in the implanted striata or in amphetamine-induced rotation between killed amnion cell-implanted and saline-injected rats did not reach significance. Implants of live amnion cells into the striatum of a parkinsonian animal model can evoke the de novo appearance of dopaminergic fibers in the denervated striatum and behavioral recovery, most likely through a trophic mechanism. PMID- 8405286 TI - Neuronal vacuolization and necrosis induced by the noncompetitive N-methyl-D aspartate (NMDA) antagonist MK(+)801 (dizocilpine maleate): a light and electron microscopic evaluation of the rat retrosplenial cortex. AB - MK(+)801 (dizocilpine maleate) is a noncompetitive antagonist at the N-methyl-D aspartate (NMDA) receptor, the major glutamate receptor at excitatory synapses in the central nervous system. Since NMDA antagonists are neuroprotective, there is interest in their development for treatment of cerebral ischemia. Unfortunately, many of these compounds also induce vacuole formation in neurons of the rat retrosplenial cortex (Olney et al., Science 244: 1360-1362, 1989). Although vacuolization was initially reported to be reversible with MK(+)801, preliminary data later suggested that higher doses might produce neuronal necrosis. To explore this issue, young male Sprague-Dawley rats were given a single subcutaneous dose of vehicle or 1, 5, or 10 mg/kg MK(+)801. At 4 h and 1, 2, 3, 4, 7, and 14 days postdose (DPD), the retrosplenial cortex was examined by light and electron microscopy. At 4 h, vacuoles occurred in neurons of retrosplenial cortical layers 3 and 4 in all rats given MK(+)801. Mitochondria and endoplasmic reticulum contributed to vacuole formation. At 1 DPD, vacuoles or necrotic neurons were rarely observed. At all subsequent time points, necrotic neurons were readily evident in rats given 5 or 10 mg/kg MK(+)801, but only rarely evident in rats given 1 mg/kg. Necrotic neurons were associated with reactive microglial cells that contained electron-dense debris ultrastructurally. If similar dose-dependent neuronal necrosis proves to be a feature of other NMDA antagonists, such effects might raise concerns for the development and use of these compounds in human cerebrovascular diseases. PMID- 8405287 TI - MR imaging of slow axonal transport in vivo. AB - Three magnetopharmaceuticals based on a monocrystalline iron oxide nanocompound (MION) are evaluated as potential contrast agents for demonstrating axonal transport in vivo by magnetic resonance (MR) imaging. One agent has a strong positive charge, one has a strong negative charge, and the third is covalently linked to wheat germ agglutinin, a plant lectin with a high affinity for axon terminals. All three agents were tagged with rhodamine, and fluorescence microscopy was used to determine their fate after administration and to validate the imaging results. Following injection into or near various neural structures in the motor and visual systems of rats, MR images were obtained at multiple times up to 11 days later, and the imaged tissues were processed for subsequent histological examination. Similar results were obtained with all three agents. Axonal transport was not seen by MR imaging or fluorescence microscopy when the agents were injected into the calf muscles, the vitreous of the eye, or the superior colliculus. However, bidirectional axonal transport was shown unequivocally by both methods after injection directly into the site of a focal crush injury to the sciatic nerve. The nerve, which otherwise is isointense with surrounding tissues on MR images, appeared as a uniformly hypointense structure having a length approximately in proportion to the time from injection to imaging. By 11 days, the course of the nerve was traceable from its component roots in the cauda equina to its bifurcation into the tibial and common peroneal nerves in the leg. A transport rate of about 5 mm/day was calculated, which is consistent with the mechanism of slow transport. MION-based magnetopharmaceuticals thus can be used to demonstrate slow axonal transport, and thereby visualize peripheral nerves, in vivo by MR imaging. PMID- 8405288 TI - Tooth pulp deafferentation is not associated with changes in primary afferent depolarization of facial afferent endings in the brain stem. AB - Previous studies have demonstrated that tooth pulp deafferentation is associated with statistically significant alterations in the low-threshold facial mechanoreceptive field properties of brain stem neurons in trigeminal (V) subnucleus oralis. A loss of spinal afferent-induced presynaptic inhibition as a consequence of a decrease in primary afferent depolarization (PAD) following spinal nerve deafferentation has been invoked as a mechanism underlying deafferentation-induced somatosensory neuroplasticity. Therefore, this study was initiated to determine if these pulp deafferentation-induced neuroplastic changes could be accounted for by an alteration in PAD of low-threshold facial afferent endings in subnucleus oralis of anesthetized rats. In control (unoperated) rats (n = 7) and rats (n = 7) that had undergone mandibular pulp deafferentation 6-10 days previously, antidromic compound action potentials evoked by test stimulation in V subnucleus oralis were recorded in branches of the infraorbital (IO) and supraorbital (SO) nerves, and conditioning stimuli were applied to some of the same nerves. PAD of the afferent endings in oralis of these nerve branches was documented in all animals, and there was no significant difference between the two groups in the incidence or any of the other features of PAD. The features of the PAD were consistent with those described in several previous studies of normal animals. These findings indicate that the reported deafferentation-induced loss of spinal presynaptic regulatory mechanisms cannot be entrapolated to all forms of deafferentation injury and that the mechanoreceptive field changes that can occur in central V somatosensory neurons as a result of tooth pulp deafferentation may not reflect an alteration in PAD. PMID- 8405289 TI - Remote microglial activation in the quinolinic acid model of Huntington's disease. AB - Intrastriatal injection of quinolinic acid (QA) in the rat leads to several structural and biochemical events which resemble neuropathological changes seen in the striatum of Huntington's disease patients. In the present experiment the accompanying microglial response in striatal projection areas following QA injection was studied immunocytochemically using monoclonal macrophage/microglial markers. After injection of 240 nmol of QA a marked microglial reaction was observed in the entire striatum, whereas injection of the same amount of solvent resulted only in a local microglial reaction around the injection site. Activated microglia were also found in the globus pallidus (GP), the entopeduncular nucleus (EP), the substantia nigra (SN), and the ventroanterior/ventrolateral, the ventromedial, and, in some rats, the reticular thalamic nucleus. The remote microglial reaction started in the first-order projection areas at Day 1 (GP) or Day 3 (EP, SN) and was found in the second-order projection areas (thalamic nuclei) by Day 5. Areas projecting to the striatum such as the amygdala and intralaminar thalamic nuclei remained free of activated microglia. It is concluded that a microglial response in striatal projection areas accompanies excitotoxic striatal injury. Anterograde degeneration of striatal projection neurons can explain the microglial activation in first-order projection areas but other mechanisms such as neuronal hyperexcitation following removal of inhibitory striatal input must be responsible for the rapid transsynaptic microglial activation seen in the thalamus. PMID- 8405290 TI - GABA agonist "muscimol" is neuroprotective in repetitive transient forebrain ischemia in gerbils. AB - The damaging effects from transient forebrain ischemia may be a result of excessive excitability or loss of inhibitory influences. In the brain, GABA acts as the major inhibitory neurotransmitter and its loss may be an important factor leading to delayed neuronal damage in the substantia nigra reticulata (SNr). In this study, we looked at the protective effects of muscimol, a GABA A agonist in a gerbil model of repetitive forebrain ischemia. For cerebral ischemia, we used three episodes of 2 min with a reperfusion period of 1 h between the insults. Histological evaluations were done 7 days after the insult using silver degeneration staining. Muscimol was infused into the third ventricle continuously for 7 days beginning just prior to the insult. There were a total of 20 animals, 12 treated with muscimol and the other 8 serving as controls. At 7 days, there was significant protection in the cortex (P = 0.007), hippocampus [CA1 (P = 0.01), CA4 (P = 0.015)], substantia nigra reticulata (P = 0.007), striatum (P = 0.049), and thalamus (P = 0.012). All statistical comparisons were done using nonparametric tests (Mann-Whitney U test). Our study shows that potentiation of inhibitory mechanisms may be important mechanisms of neuronal protection from the effects of repetitive ischemia and the effects are not limited to the SNr. Further studies are needed to better understand their mechanism of action. PMID- 8405291 TI - Differential increases in catecholamine metabolizing enzymes in amyotrophic lateral sclerosis. AB - The activity of three catecholamine-metabolizing enzymes, monoamine oxidase type A and type B (MAO-A and MAO-B) as well as catechol-O-methyltransferase (COMT), were estimated in homogenates of human spinal cord using radiometric assays. The enzyme activities were determined in postmortem spinal cord tissue from controls and cases with amyotrophic lateral sclerosis (ALS). The activity of MAO-A was below the limit of detectability in both controls and ALS cases. The activities of MAO-B and COMT were evenly distributed at the various spinal levels. The MAO-B activity was substantially elevated in ALS spinal homogenates, whereas only a slight, but not statistically significant, increase in COMT activity was observed. A significant correlation between COMT and MAO-B activities was observed for controls. However, this covariation was not apparent for the ALS cases. These results suggest that the two enzyme proteins are regulated by more complex mechanisms in the spinal cord in amyotrophic lateral sclerosis than simple general increases caused by elevated astroglial cell numbers. In addition, the MAO-A, MAO-B, and COMT activities were estimated in spinal cords from rats treated with the selective MAO-B inhibitor L-deprenyl, a drug with putative neuroprotective effects in neurodegenerative disorders. After 3 weeks of L deprenyl treatment (0.25 mg/kg/day, sc), the spinal MAO-A and MAO-B activities were decreased by 50 and 80%, respectively. In contrast, the COMT activity was not altered by L-deprenyl administration. PMID- 8405292 TI - Pharmacological induction of nerve growth factor mRNA in adult rat brain. AB - Three structurally unrelated compounds, all of which induce nerve growth factor (NGF) in cell culture systems, were assessed for their ability to induce NGF mRNA in adult rat brain using a highly sensitive RNAse protection assay. Interleukin-1 beta (0.5-1 pmol) and 1,25-dihydroxyvitamin D3 (25-25,000 pmol) were extremely potent inducers of NGF mRNA, being respectively at least 50,000 and 4000 times more potent than 4-methylcatechol. These compounds elicited an approximate twofold increase in NGF mRNA in both the hippocampus and cortex, without altering beta-actin mRNA levels after a single intracerebroventricular injection. The duration of NGF induction was dependent on the compound administered. For example, the elevation of NGF mRNA elicited by interleukin-1 beta peaked at 8 h and lasted for at least 24 h. In contrast, the induction of NGF after 1,25 dihydroxyvitamin D3 and 4-methylcatechol administration peaked between 4 and 8 h and was not apparent 24 h after injection. These results demonstrate induction of NGF mRNA in vivo by administration of physiological or pharmacological agents and differentiate these agents by potency and duration of action. Further, these findings indicate that pharmacological induction of NGF may be a viable strategy for the treatment of neurodegenerative disorders such as Alzheimer's disease. PMID- 8405293 TI - Selective reinnervation of the rat serratus anterior muscle following denervation and partial target removal. AB - The rat serratus anterior (SA) muscle is reinnervated by cervical roots C6 and C7 with a topographic bias following transection or freezing of the long thoracic nerve. The study reported here was undertaken to determine the specificity of regenerating motoneurons when the size of the target SA muscle was reduced. After crush and removal of the caudal half of the muscle, a rostrocaudal map was reestablished that was no different from control or crush alone. Rostral (C6) axons dominated the innervation of muscles deprived of their caudal sectors. Input from caudal (C7) axons was significantly reduced. These results may be explained by preferential pathways or a competitive advantage held by rostral axons reinnervating rostral sectors to the detriment of caudal axons. PMID- 8405294 TI - The atmospheric environment--an introduction. AB - The atmosphere is part of the environment with which the human organism is permanently confronted. Epidemiological research investigates the occurrence of effects on morbidity and mortality due to heat, cold, air pollution and changes in the weather. Concentrating on aspects of the environment relevant for medical questions, three major complexes of effects can be discriminated: the complex conditions of heat exchange, the direct biological effects of solar radiation, and air pollution. Biometeorological knowledge can serve to assess the atmospheric environment, and can also be of help in the field of preventive planning, to conserve and develop the climate as a natural resource with regard to man's health, well-being and performance. PMID- 8405295 TI - Heat balance modelling. AB - The only way to describe the effects of the thermal environment on the human body completely is to do it by means of an energy balance equation. In such an equation all relevant meteorological parameters, behavioral characteristics (activity and clothing) and body measurements can be considered. Using Fanger's comfort equation and the models MEMI and IMEM as examples, the problems of energy balancing and ways of solving them are described. The value of energy balance models is documented by examples from the field of application. PMID- 8405296 TI - Biological effectiveness of solar UV radiation in humans. AB - Solar UVB radiation is prejudicial to the health of humans in a number of ways. Erythema and photodermatoses are acute reactions of the skin; keratitis and conjunctivitis are acute reactions of the eye. Various types of skin cancer, accelerated aging of the skin, and cataract formation in the crystalline lens are reactions that appear with great latency. UV radiation can also cause damage to the immune system and DNA. For the period 1981-1991, an increase in erythemal effective UVB radiation of +(7 +/- 4)% per decade was measured in a non-polluted high mountain area (Jungfraujoch, 3576 m a.s.l., Switzerland). This increase is related to a decrease in stratospheric ozone. The effects on human health are discussed. A 10% ozone reduction increases non-melanoma skin cancer by 26% and cataract by 6 to 8%. PMID- 8405297 TI - Effects of atmospheric pollution on human health. AB - Most air pollutants do not lead to specific diseases. Depending on the pollutant, the concentration and the duration of exposure, some organs are more affected than others. The most frequent disorders are those caused by irritant gases and particulates on the mucous membranes and respiratory organs. The consequences are eye, nose and throat inflammations, diminished lung function, increased susceptibility to respiratory infection and a higher incidence of chronic bronchitis. These disorders and diseases are, of course, influenced by other factors as well, such as immune deficiency, allergies, occupational exposure to pollutants, and particularly smoking. The effects of air pollutants are, therefore, multifactorially conditioned and nonspecific disorders are placed in the foreground. Evidence for an association of air pollution with adverse effects on human health is drawn from three sources: animal experiments, experimental human exposures, and epidemiologic studies of exposed human populations. The burden of atmospheric pollution must be reduced to protect human health by an adequate safety margin. In particular, the increased sensitivity of sick and aged people as well as children should be taken into account. In defining the maximum emission levels, preventive aspects should have priority so as to keep the risk of damage to health and the harmful influences on the environment to a minimum. PMID- 8405298 TI - Meteorotropy and medical-meteorological forecasts. AB - The meteorotropic reaction of the human organism is a function of different factors, such as the type and intensity of the effects of the physical environment as well as individual conditions like adaptive capacity and state of health. Many medical-meteorological studies show causal correlations between conditions in the lower atmosphere and reactions of the human organism, but also combined or synergistic effects of different weather situations, which can only be proved stochastically. These effects are described and the methodology of the investigations, as well as the results, critically discussed. Furthermore, application of the results in the areas of medical-meteorological consultation, with the goal of improving living conditions, is considered. PMID- 8405299 TI - Possible biological effects of electric and magnetic parameters in the environment. AB - The features and intensities of electric and magnetic fields in the environment are described. Natural as well as technical constant and variable fields are considered in the light of their possible biological effects. The upper limits of the various fields are discussed. Results of laboratory measurements and also of epidemiological investigations are presented. The question is raised whether atmospheric small ions can cause a biological effect. PMID- 8405300 TI - Indoor climate. AB - In industrialized countries most of the time is spent indoors. The basic ambient parameters for a thermally comfortable indoor climate are air temperature, air velocity, humidity and radiation pattern. Besides the thermal component, the concentrations of air pollutants in the indoor air are also of importance for wellbeing and health. Their levels are influenced both by the outdoor concentrations and the indoor emissions. The increasing use of air conditioning systems in many cases has not resulted in improving the indoor climate but causes a wide range of irritations and health problems summarized as 'sick building syndrome'. PMID- 8405301 TI - The role of histones and their modifications in the informative content of chromatin. AB - It is traditionally accepted that the DNA sequence cannot by itself explain all the mechanisms necessary for the development of living beings, especially in eukaryotes. Indeed part of the information used in these processes is stored in other ways, generally called 'epigenetic', whose molecular mechanisms are mostly unknown. The ultimate explanation for them might reside in the non-DNA moiety of chromatin which may play an active role in heredity ('chromatin information'). Histones are the universal structural component of chromatin. However, recent studies strongly suggest that histones, and their modifications--especially the reversible acetylation of lysines--may act as a recognition signal for regulatory proteins and they may participate, for this reason, in gene regulation. This type of information could be maintained through its replication and, ultimately, it could form the molecular basis of certain processes related to the development of the eukaryotic organisms. PMID- 8405302 TI - Carbonic anhydrase type II in regenerating retinal pigment epithelium. A histochemical study in the rabbit. AB - Type II carbonic anhydrase (CAII) in the cytoplasm of the retinal pigment epithelium (RPE) may contribute to the transport of water and solutes across the RPE. The activity of this enzyme in RPE during its response to damage, e.g., during regeneration, is therefore of interest in understanding retinal disease. Immuno-histochemistry was used to compare CAII activity of normal RPE and RPE experimentally induced to regenerate. In normal rabbits, the RPE stained intensely with a peroxidase-linked antibody specific for human CAII. Regenerating RPE stained less intensely. Within the regenerating epithelium, staining appeared more intense in mature cells than in immature ones, suggesting that CAII activity gradually returns during RPE regeneration. PMID- 8405303 TI - Involvement of thiols in the induction of inward current induced by silver in frog skeletal muscle membrane. AB - Exposure of voltage-clamped frog skeletal muscle fibres to silver caused a maintained inward current which could be carried by Ca2+, Mg2+ or Na+. Inorganic Ca2+ channel blockers and dithiothreitol (SH reducing agent) diminished this current, but a Na+ channel blocker did not. Thus, silver activates the Ca2+ channel by acting on SH groups in a Ca2+ channel protein. PMID- 8405304 TI - Hepatic glucose signals vagally modulate the cyclicity of gastric motility in rats. AB - The cyclicity and intensity of gastric motility were examined following glucose injection into the portal vein with an intragastric balloon in anesthetized rats. Enhanced gastric motility caused by insulin administration was influenced by 4 mM glucose (25 microliters) injected into the portal vein; glucose provoked a shift in the cyclicity power spectrum without any change in intensity. The peak power spectrum shifted from 4.0-5.0 cpm to 2.0-3.0 cpm. Hepatic branch vagotomy abolished the response. The results suggest that glucose signals in the hepatic vagal branch modulate the cyclicity of gastric motility. PMID- 8405305 TI - Similarity between the effects of suprachiasmatic nuclei lesions and of pinealectomy on gonadotropin release in ovariectomized, sulpiride-treated and melatonin-replaced rats. AB - The aim of this study was to compare the effects of pineal indole treatments on LH and FSH release in pinealectomized and suprachiasmatic lesioned and ovariectomized rats rendered hyperprolactinemic by acute sulpiride treatment. Pinealectomy or suprachiasmatic nuclei lesions in female rats both decreased plasma LH and FHS at 10, but not at 20 d after surgery, whereas the daily afternoon administration of melatonin effectively restored levels of both gonadotropins to control values. In ovariectomized rats, pinealectomy or suprachiasmatic nuclei lesions were ineffective in counteracting the high plasma levels of LH and FSH. However, sulpiride treatment in both pinealectomized and suprachiasmatic nuclei lesioned and castrated female rats significantly decreased the levels of LH and FSH, an effect which was counteracted by daily afternoon melatonin administration. Other pineal indoles tested, i.e., 5-hydroxy- and 5 methoxytryptophol, were ineffective in regulating gonadotropin levels. The results suggest that the pineal gland, through its hormone melatonin, can modulate gonadotropin secretion by acting on a dopamine mechanism independent of hypothalamic suprachiasmatic areas. PMID- 8405306 TI - Non-viraemic transmission of tick-borne encephalitis virus: a mechanism for arbovirus survival in nature. AB - The vectors of arthropod-borne viruses (arboviruses) become infected by feeding on the viraemic blood of an infected animal. This theory is based on transmission studies involving artificial infection of vertebrate hosts by syringe inoculation. To reproduce natural conditions of virus transmission, infected and uninfected vectors (ticks) of tick-borne encephalitis virus, the most important arbovirus in Europe, were allowed to feed together on uninfected wild vertebrate hosts. The greatest numbers of infected ticks were obtained from susceptible host species that had undetectable or very low levels of viraemia. The results suggest that 'nonviremic transmission' is an important mechanism for the survival of certain arboviruses in nature. PMID- 8405307 TI - The structure of scytonemin, an ultraviolet sunscreen pigment from the sheaths of cyanobacteria. AB - Despite knowledge of the existence of the pigment called scytonemin for over 100 years, its structure has remained unsolved until now. This pigment, the first shown to be an effective, photo-stable ultraviolet shield in prokaryotes, is a novel dimeric molecule (molec. wt. 544) of indolic and phenolic subunits and is known only from the sheaths enclosing the cells of cyanobacteria. It is probable that scytonemin is formed from a condensation of tryptophan- and phenylpropanoid derived subunits. The linkage between these units is unique among natural products and this novel ring structure is here termed the 'scytoneman skeleton'. Scytonemin absorbs strongly and broadly in the spectral region 325-425 nm (UV-A violet-blue, with an in vivo maximum at 370 nm). However, there is also major absorption in the UV-C (lambda max = 250 nm) and UV-B (280-320 nm). The pigment has been recently shown to provide significant protection to cyanobacteria against damage by ultraviolet radiation. The pigment occurs in all phylogenetic lines of sheathed cyanobacteria and possibly represents a UV screening strategy far more ancient than that of plant flavonoids and animal melanins. How diverse organisms deal with UV radiation is considered of vital importance to global ecology. PMID- 8405308 TI - Concurrent antigenic analysis of recent epidemic influenza A and B viruses and quantitation of antibodies in population serosurveys in Italy. AB - Laboratory investigations of virus isolation and serum antibodies in a Mediterranean country (Italy) demonstrated that influenza A and B viruses, and often both, circulated every winter in Italy. The winter 1987/88 was characterized by a low level of influenza activity, as shown by the limited number (47) of influenza virus isolates, the majority of which (61%) belonged to the influenza B type. In contrast, the 1988/89 influenza season was exclusively associated with the circulation of influenza type A viruses. The A(H1H1) subtype was largely predominant (97%), as compared to the low incidence of the A(H3N2) subtype (3%). During the 1989/90 winter a co-circulation of A and B influenza viruses was observed, A(H3N2) strains being responsible for 96% of the virologically confirmed cases. Antigenic analysis of the virus isolates showed some antigenic variation in influenza A viruses of both H1N1 and H3N2 subtypes, whilst antigenic stability was found among the influenza B virus isolates. Overall, the above virological findings correlate with the data concerning the pattern of influenza virus circulation in Northern Europe and the UK during the three years surveyed. The results of serum antibody surveys conducted in each post-epidemic period are also reported. PMID- 8405309 TI - Investigation of an outbreak of Salmonella typhi in a public school in Madrid. AB - A typhoid fever outbreak affecting 54 school students occurred in a Public School of Mostoles, Madrid. The date of onset was 11 June 1991 and the last detected case was 8 July 1991. Salmonella typhi was cultured from blood and/or stool samples corresponding to 54 patients and one food-handler. There were no secondary cases detected. Epidemiological investigation suggested a salad or a custard as the common source. Patients and the food-handler were treated with ampicillin/amoxicillin for up to three weeks. There were seven relapses that were also treated with the same antibiotics with success. None were found to be excreting the organisms when tested after four months. All the Salmonella typhi isolated strains were phagetype 34, biotype Xylose +, Tetrationate Reductase + and harboured a similar 22 Mdal plasmid, they were also susceptible to the antibiotics tested. PMID- 8405310 TI - A longitudinal study on the incidence and transmission patterns of HIV, HBV and HCV infection among drug users in Amsterdam. AB - In the present study data on the incidence of HBV and HCV were used to indicate the prevalence of and trends in risk behavior, assuming that drug users (DUs) who become infected with HBV or HCV are also at risk for infection with HIV. In addition, we determined to that extent the transmission patterns of HIV, HBV and HCV differed. DUs were selected from a cohort study in Amsterdam, had at least one follow-up visit between December 1985 and September 1989 and reported never to have had homosexual contacts. Among 305 DUs, of whom 70% injected recently, the prevalence of HIV, HBV and HCV were 31%, 68% and 65% respectively. These prevalences were strongly interrelated and the same risk factors were found. The cumulative incidence of either HIV or HBV or HCV was 30% among prevalent HIV negatives. Despite a previously reported reduction in risk behavior, only the HIV incidence tended to decrease initially, and after 1986 the incidences of HIV, HBV and HCV remained disturbingly high and stable (mean: 4, 9 and 10 per 100 person years, respectively). As at present HBV appears to be transmitted more heterosexually than HIV in our study group and the HIV-epidemic may follow the HBV-epidemic in its transmission patterns, preventive activities targeted at both injecting and sexual behavior should be expanded. PMID- 8405311 TI - Validity of the cross-sectional study for the ascertainment of nosocomial infection risk factors. AB - This study was carried out in order to assess the validity of the pure cross sectional study in the ascertainment of nosocomial infection risk-factors. The results yielded by two designs (cross-sectional and case-control) are compared. A cross-sectional design was performed in a tertiary hospital. 592 patients were studied, 38 of whom were nosocomially infected. The clinical information on all the patients included in this design was reviewed after hospital discharge. A matched case-control study was nested in the population cross-sectionally surveyed. 66 cases (28 additional patients developed a hospital infection) and 132 controls were selected. Odds ratios (ORs) for the risk factors analyzed by both designs were compared. There were no significant differences between the estimates yielded by both designs; however, a trend of lower OR estimates for the cross-sectional study was seen, which may be important for risk factors not strongly related to (low relative risk) nosocomial infection. Several factors which might account for the results observed (random error, bias introduced by matching) are discussed. It is suggested that pure cross-sectional designs for the study of risk factors of nosocomial infection may introduce a negative (toward-the-null) bias. PMID- 8405312 TI - Comparison of western blot and microimmunofluorescence as tools for Lyme disease seroepidemiology. AB - Sera from a population of 212 farmers of the Puy de Dome (Midlands of France) who are in close contact with Ixodes ricinus, the vector of Lyme disease in Europe, and sera from 100 urban blood donors from the nearly city of Clermont Ferrand were examined by microimmunofluorescence (MIF) for antibodies reacting with Borrelia burgdorferi. The results showed a higher seroprevalence of IgG > 1/100 in farmers (25%) than in blood donors (10%). Using western blot with antibody at a 1/200 dilution, and regarding sera as being positive if they contained IgG reactive with more than 5 bands, 93/212 farmers (44%) and 20% of blood donors were positive. Reactions with specific protein bands (94, 73, 30 and 21 KDa) by western blot were demonstrated in 83/212 sera from farmers (39%) and 16% of blood donors. Both methods showed a higher seroprevalence in the farmer population. Western blot is a sensitive and specific test for seroepidemiology but, in highly endemic areas, it is not diagnostic for active Lyme borrelliosis. PMID- 8405313 TI - The incidence of acquired immune deficiency syndrome (AIDS) and tuberculosis (TB) in the Hospital "La Fe" of Valencia, Spain (1985-1989). AB - We studied the incidence of TB and TB-AIDS in the area served by "La Fe" hospital in Valencia. We also studied the different evolution of the incidence of TB and TB-AIDS during the 1985-1989 period. We noticed the progressive increase of the incidence of TB in AIDS patients, while the incidence of TB without AIDS remained the same. PMID- 8405314 TI - Cardiovascular risk and all-cause mortality; a 12 year follow-up study in The Netherlands. AB - To assess the contribution of cardiovascular risk indicators to all-cause mortality, we used data from a follow-up study conducted in the Netherlands since 1975. Of 6,057 participants aged 20 years or over at the start of the study, 9.5% died during the 9 to 12 year follow-up period. Risk indicators independently related to all-cause mortality were age and diabetes mellitus in both sexes; pulse rate, smoking habits, antihypertensive drug use and a history of myocardial infarction most clearly in men; and body mass index and systolic blood pressure in women. A larger body mass index was associated with a gradual decrease in mortality probability. The risk of death for women in the highest quartile of body mass index (> 26.4 kg/m2) relative to those in the lowest quartile (< 21.9 kg/m2) was 0.56 (95% confidence limits 0.36 and 0.87). Serum cholesterol level showed no association with overall mortality. Risk functions were calculated to predict an individual's probability of dying within 11.5 years as a function of the level of cardiovascular risk indicators. Our findings suggest that the major cardiovascular risk indicators, apart from affecting cardiovascular morbidity and mortality, also influence all-cause mortality. Consequently, favourable changes in these characteristics might lead to an increase in life expectancy. The maximum individual benefit to be expected from these changes may be estimated using the risk functions derived from our data. PMID- 8405315 TI - Relations of cardiovascular risk factors to aortic pulse wave velocity in asymptomatic middle-aged women. AB - Cross-sectional associations between aortic elasticity assessed by carotid to femoral pulse wave velocity (PWV) and cardiovascular risk factors were examined in 429 apparently healthy middle-aged women. PWV was strongly and positively related to blood pressure and hypertension. Weak but significant positive associations were also found between PWV and age, heart rate, some lipids and lipoproteins, blood glucose (either as a continuous or dichotomous variable), body mass index, haematocrit, leucocyte count and family history of diabetes. No associations were observed between PWV and high density lipoprotein cholesterol, apolipoprotein A1, fibrinogen, cigarette smoking, menopausal status and a family history of hypertension or myocardial infarction. After adjustment for systolic blood pressure, PWV remained significantly related to heart rate, leucocyte count, blood glucose (as a dichotomous variable) and a family history of diabetes. Multiple regression analysis showed that systolic blood pressure and, to a lesser extent, heart rate, leucocyte count and a family history of diabetes were all independent determinants of PWV. This pattern of associations suggests that arterial stiffness measured by PWV reflects the sclerotic rather than the atherotic component of atherosclerosis. The potential influence of a family history of diabetes on the elastic properties of the aorta needs to be ascertained in further studies. PMID- 8405316 TI - Hepatitis B mass immunization of adolescents: a pilot study in a community. AB - The National Type Specific Hepatitis Surveillance System (SEIEVA) and seroepidemiological studies have shown that in addition to newborns from mothers who are carriers for hepatitis B surface antigen (HBsAg), adolescents are at high risk of acquiring type B hepatitis virus because of increasing importance of the heterosexual transmission of this virus. In order to evaluate logistic problems and acceptance rate of adolescents to mass vaccination against hepatitis B, a pilot study was carried out among all 7th grade children registered in the 9 schools of an hepatitis B endemic area located in the suburbs of Naples. After meetings held by the local health department with school teachers and parents, 1219 out of 1250 (97.5%) invited children received the first dose of hepatitis B vaccine; 1215 and 1209 received, respectively, the second and third doses. Anti hepatitis B surface antigen (anti-HBs) values were studied in 406 subjects one month after the third dose, and 21 out of 406 (5.2%) had anti-HBs values less than 10 IU/L. We consider 95% of subjects with anti-HBs values greater than 9 IU/L a good achievement for a field vaccination program. The high acceptance rate of vaccination found in our study outlines the importance of active offer of vaccination combined with school involvement. PMID- 8405317 TI - Duration of the immune response to MMR vaccine in children of two age-different groups. AB - A combined vaccine against measles, mumps and rubella (MMR) was administered to both a group of children aged 10-12 months simultaneously with booster doses of compulsory diphtheria-tetanus toxoids and oral poliovirus vaccine and a group of children aged 15-24 months who had previously received booster doses of the compulsory vaccines. Apart from one subject belonging to the second group who was non responder and one from the same group who did not seroconvert against the mumps virus alone, 5 to 6 weeks after MMR vaccine administration we found protective levels of antibodies against measles, mumps and rubella viruses in all children. The follow up of both groups at 3 years did not reveal difference between the two groups. Protective levels of serum antibodies against measles and mumps were found in the two groups, although a significant decline of rubella antibodies was shown (p < 0.05). Since the immunogenicity of the vaccines in the two groups did not differ, we recommend that the scientific community reconsider the vaccination schedule until now recommended. In our opinion the MMR vaccine should be administered simultaneously with booster doses of diphtheria-tetanus toxoids and oral poliovirus vaccine at 10-12 months of age because this policy improves parents' compliance, markedly reduces community costs and simplifies routine immunization schedule. PMID- 8405318 TI - Comparisons between degree of histological gastritis and DNA fingerprints, cytotoxicity and adhesivity of Helicobacter pylori from different gastric sites. AB - Thirty-six isolates of H. pylori from up to three gastric biopsy sites (antrum, corpus and fundus) from 13 patients in Italy with different degrees of histological gastritis were investigated. All strains were tested for motility, cytotoxicity and degree of adhesion, and were typed by analysis of ribosomal RNA gene patterns (ribopatterns). Seventeen different DNA types (ribotypes) were identified, with each patient possessing H. pylori of one or more unique types. Only two patients had identical H. pylori at three sites. Most patients had H. pylori with different ribotypes or subtypes, but nine strains were not typable. Five patients had the same strain colonizing two of the three sites and atypical strains were mostly from the antrum. A complex pattern of H. pylori colonization in the stomach of some individuals was evident and suggested multiple sources of infection. No consistent associations were detected between degree of gastritis and adherence, cytotoxicity and motility but a 2.56Kb rRNA gene fragment that had a higher frequency in strains associated with severe gastritis than mild gastritis, may provide a useful molecular marker for future pathogenicity studies. PMID- 8405319 TI - Seroepidemiology, morbidity and vaccination strategies against rubella infection. Eight years experience in Oltrepo Pavese. AB - Selective rubella vaccination of schoolgirls in Italy started 14 years ago following the United Kingdom strategy that was adopted in 1970. The aims of this program were to eliminate the risk of rubella among women of childbearing age, encourage the acquisition of immunity by natural infection during early childhood and allow the vaccine-induced antibody production by the circulating virus. On the basis of this program, between 1982 to 1990, a prospective serosurvey for rubella antibody in the province of Pavia was performed. The results showed a decline in the overall seropositivity rate for rubella antibodies from 57.7% in 1982 to 41.9% in 1984 followed by a remarkable increase in 1985 (53.3%) and in 1987 (56.5%). This trend was confirmed by the number of cases reported to the local Public Health Service. The results of this study provide further evidence of the need to change the current selective immunization policy in order to obtain a significant reduction of risk of the infection in the population. PMID- 8405321 TI - Urinary tract infections in the city of Florence: epidemiological considerations over a twenty-year period. AB - Our study of significant bacteriurias indicated that the worldwide shift in the etiology of infections also holds true for the Florence area. In a twenty-year period (1970-1990), we noted a decreased frequency of Gram-negative bacilli, particularly of the family Enterobacteriaceae, and a significant increase of Gram positive cocci in urinary patients. This finding was observed both in hospital and in community-acquired cases in the male sex and only in nosocomial bacteriurias in the female sex. There was a reduced isolation of "classic" urinary pathogens such as Proteus mirabilis: its prevalence in hospital-acquired urinary tract infection (UTI) decreased from 16% in 1970 to 5% in 1990 both in males and in females. On the other hand, we noted an increase of "difficult" microorganisms such as enterococci and methicillin-resistant staphylococci, particularly in the male sex; in 1970 enterococci were occasionally isolated in males both from hospital and from community-acquired UTIs (3% and 5%, respectively), whereas in 1990, on the contrary, they were encountered much more frequently (19% in both cases). PMID- 8405320 TI - Improvement of Clostridium difficile isolation by heat-shock and typing of the isolated strains by SDS-PAGE. AB - Clostridium difficile plays an essential role in causing pseudomembranous colitis. We looked for the presence of these bacteria in the stools of 169 hospitalized patients and 38 nurses from wards with cases of diarrhea (207 subjects). The study was divided into three parts. In the first part, we compared three methods for isolating Clostridium difficile from stool samples: pre selection with heat-shock, direct plating on Cycloserine-Cefotaxime-Fructose Agar (CCFA) and culturing in a selective broth medium. Final identification of Clostridium difficile was achieved by gas-chromatography and ApiZym. From the 207 consecutively obtained stool specimens, Clostridium difficile was isolated in 108 (52%) when pre-treated by heat-shock compared to only 26 (13%) when plated on modified CCFA and 23 (11%) when cultured in selective broth medium. Pre-selection significantly increases the isolation rate for Clostridium difficile and should be used in further epidemiological research. In the second part of our study, a retrospective review of subjects' records showed that the heat-shock method detected Clostridium difficile in all age groups at a higher rate than the other methods. In the third part of our study, we typed the 157 isolates of Clostridium difficile strains by protein patterns using SDS-PAGE, and 16 distinct groups were identified. In 19 cases different Clostridium difficile strains were found in the same subject by SDS-PAGE. Finally, the isolated strains were compared with strains from Brussels and Freiburg. Matching patterns were noted in only three cases. PMID- 8405322 TI - Seroprevalence to some TORCH agents in a Sicilian female population of fertile age. AB - The seroprevalence to Toxoplasma gondii (41.1%), rubella virus (88.2%), cytomegalovirus (86.0%), and herpes simplex virus (80.0%) has been evaluated in fertile women living in Catania (Sicily). The population group studied was divided into four age groups to quantify the risk of primary infection in each age group. PMID- 8405323 TI - Antibodies to Rickettsia conorii in dogs: seasonal differences. AB - Eight dogs, having showed positivity to Rickettsia conorii in serum samples obtained during the spring and summer, were studied again by means of a second determination during the next winter, 4-10 months later. Serum titer became negative in six dogs, persisted high in one, and fell from 1:640 to 1:40 in another dog. This seasonal difference suggests a short persistence of antibodies in dogs after contact with R. conorii in the Mediterranean area. PMID- 8405324 TI - Seroepidemiological survey of HTLV-I infection in French Polynesia, Cook Islands and Fiji. AB - Different population groups of French Polynesia, Cook Islands and Fiji were screened for Human T-Lymphotropic Virus type I (HTLV-I) antibodies. Among 1487 individuals sampled in French Polynesia, twelve were considered Western Blot (WB) indeterminate and one was considered WB-positive for HTLV-I infection. This positive subject originated from France and was a blood donor. Out of 196 Polynesians of the Cook Islands, one was WB-indeterminate. Among populations sampled in Fiji, one of 222 Melanesians was found WB-indeterminate and one of 211 Indians was WB-indeterminate. PMID- 8405325 TI - Prevalence of HTLV-II in HIV-1-infected drug addicts in Marseille. PMID- 8405326 TI - Apparent polycythaemia: diagnosis, pathogenesis and management. AB - The term "apparent polycythaemia" is applied to a group of patients who have a raised PCV (> 0.51 in males, > 0.48 in females) but a normal red cell mass (less than 25% above their predicted mean normal value). Some have additionally a marked reduction in plasma volume and can be defined as a subgroup: relative polycythaemia. Smoking, hypertension and to a lesser extent obesity, excessive alcohol, low-dose diuretic therapy and hypoxaemia have all been associated with apparent polycythaemia but the mechanism is both uncertain and likely to be complex. This group of patients is unlikely to be uniform in pathogenesis and may well include some normal individuals. Investigation requires exclusion of factors associated with other types of polycythaemia. The possibility of an increased vascular occlusive risk is uncertain in these patients except at the higher PCV values. Reduction of PCV by venesection is sensible at PCV > 0.54 or where there is perceived to be an increased risk of vascular occlusion. The remaining patients should be managed by regular observation to detect further rise in PCV or evolution to absolute polycythaemia (raised red cell mass). In some, the PCV returns to normal. PMID- 8405327 TI - Epidemiological and clinical study of sickle cell disease in France, French Guiana and Algeria. AB - The main clinical and haematological features of sickle cell patients were compared in 618 French, 50 Guianese and 87 Algerian patients. In homozygous sickle cell patients, the proportion of icteric subjects rises with age in all centres; the prevalence of splenomegaly reaches a peak in children from 1 to 5 years and then decreases; jaundice and splenomegaly are more often noted in Algerian and Guianese than French patients. The prevalence of painful crisis is comparable in the 3 centres. In 465 French SS children, having a mean age of 7.3 +/- 5.9 years, the prevalence of a past history of meningitis is 7.3%, of septicaemia 4.1% of osteomyelitis 8.8%. These percentages do not differ significantly between countries. Prevalence of a past history of cerebrovascular accident is 3.2% in French SS patients; 1.2% in SC, 3.8% in S beta thalassaemia. A past history of acute splenic sequestration was noted significantly more often in SS (11.75%) and S beta thalassaemia (14.3%) than SC (3.6%) in French children (p < 0.05). Proportions of subjects transfused at least once do not differ between countries; SS children are more transfused (64%) than SC (15.6%) and S beta thalassaemic (66%) (p < 10-4). Haemoglobin and reticulocyte counts do not differ significantly between countries. In conclusion, no major differences were detected between French, Guianese and Algerian homozygous sickle cell patients: this may be due to the fact that France is in itself a mosaic of ethnic origins. PMID- 8405328 TI - Measurement of spleen size using gamma camera scintigraphy in essential thrombocythaemia. AB - By using gamma camera imaging the spleen size was determined in 33 consecutive patients with essential thrombocythaemia (ET) and in 33 consecutive patients with reactive thrombocytosis (RT). All ET patients were newly diagnosed and had not received myelosuppressive treatment prior to study; they all fulfilled the criteria for ET as established by the Polycythemia Vera Study Group. In both posterior and lateral projections, the spleen area in the group of ET patients was significantly larger than in the RT patients. The present study has shown that 39% of ET patients at diagnosis have splenic enlargement. Evaluation of spleen size is therefore a useful diagnostic test in patients presenting with unexplained thrombocytosis. PMID- 8405329 TI - Functional capacity of transfused platelets estimated by the Thrombostat 4000/2. AB - Post-transfusion platelet increment is a useful test for the evaluation of transfusion efficacy. It does not, however, give information about the in vivo platelet function. We have evaluated the in vitro bleeding time (IVBT) using the Thrombostat 4000/2 with ADP as activating agent in 60 platelet transfusions given to 17 chemotherapy-induced severely thrombocytopenic patients. Determinations were performed before and after transfusion of platelet concentrates (PC) prepared from buffy coat. Both fresh and stored platelets resulted in significant reductions of the IVBT already 10 minutes after completed platelet transfusion. In the majority of patients there was a correspondence between IVBT and platelet increment. However, in 30% of the cases there was no improvement of the IVBT despite an increase of the platelet count. Fresh PCs were more effective than stored. The IVBT seems to represent a clear-cut improvement in the possibilities for evaluating and monitoring the effect of platelet transfusion. PMID- 8405330 TI - Improved survival from fungaemia in patients with haematological malignancies: analysis of risk factors for death and usefulness of early antifungal therapy. AB - Fourty-three episodes of fungaemia encountered from 1978 to 1991 in 43 patients with haematological malignancies are reviewed here to analyse the risk factors for death and to evaluate the efficacy of early antifungal therapy. Low serum cholinesterase and elevated serum blood urea nitrogen were significantly associated with fungaemic death, defined as death occurring within 2 weeks after documentation of fungaemia. Overall death rate from fungaemia was 62.8%. Before the introduction of early antifungal therapy in 1986, however, fungaemic mortality was 85.7%; it was reduced to 51.7% thereafter (p = 0.01). Determination of plasma (1-->3)-beta-D-glucan was helpful in detecting deep fungal infections and initiating antifungal therapy early. PMID- 8405331 TI - Neutrophils from patients with secondary haemosiderosis contain excessive amounts of autotoxic iron. AB - Secondary haemosiderosis may be accompanied by a decrease in the phagocytic function of neutrophils (PMNs). This dysfunction has been attributed to an exaggerated generation of oxidants induced by intracellular iron. However, an accumulation of iron has so far not been reliably demonstrated in neutrophils harvested from iron-overloaded patients. Six polytransfused haemodialysed patients, with a serum ferritin level higher than 1000 micrograms/l, and 10 healthy controls were investigated. The iron status of PMNs was evaluated by iron determination using atomic absorption spectrometry and by ferritin measurement using radioimmunoassay. The phagocytic performance was measured by cytofluorometry. The results confirm that PMNs from the haemosiderosis patients have a decreased phagocytosis. Moreover, they demonstrate for the first time that these PMNs have an increased cellular iron and ferritin content. Both latter concentrations were 4 to 5 times more elevated in secondary haemosiderosis than in healthy controls. This iron accumulation may be toxic for the PMNs and may, at least partially, explain the three-fold higher risk of bacteraemia which has been reported in those patients. PMID- 8405332 TI - Prolonged hematological remission by mitoxantrone and interferon--alpha-2b in a child with resistant Philadelphia chromosome-positive chronic myelogenous leukemia. PMID- 8405333 TI - Association between haemoglobin and neopterin concentrations in haemodialysis patients treated with erythropoietin. PMID- 8405334 TI - Three years' continuous low-dose interferon-alpha treatment of hairy-cell leukemia: evaluation of response and maintenance dose. PMID- 8405335 TI - Interaction between cyclosporin and chlorambucil. PMID- 8405336 TI - Chemotherapy of acute myeloblastic leukemia in an HIV carrier. PMID- 8405337 TI - Adult respiratory distress syndrome complicating hematological diseases. PMID- 8405338 TI - Iron deficiency in sickle cell anemia. PMID- 8405339 TI - The ileocecal syndrome in acute leukemia--is thrombosis an important part of pathogenesis? PMID- 8405340 TI - Educational outcomes in teenage pregnancy and parenting programs: results from a demonstration. AB - A comparison of five in-school educational and service approaches offered at seven sites in Arizona to 789 pregnant and parenting teenagers shows that except for those who enroll in a program in their third trimester, pregnant and parenting teenagers who attend a comprehensive, school-based, community-linked program are significantly more likely to continue in school than are those who have no access to a special program. The comprehensive program's impact is greatest among Hispanic students, younger students, those in grades 9-10, those who are living with their partner and those who enter the program in the first trimester. Two of the program components--strong outreach efforts and case management-are believed to have an especially favorable impact on continuation in school. PMID- 8405341 TI - The socioeconomic consequences of teenage childbearing: findings from a natural experiment. AB - A study based on census data from 1970 and 1980 examines the socioeconomic effects of unplanned teenage childbearing by comparing teenage mothers whose first birth was to twins with those whose first birth was to a single infant. Among black women, an unplanned teenage birth--represented by the secondborn twin -results in significantly lower rates of high school graduation and labor-force participation and significantly higher rates of poverty and welfare recipiency. Ten years after giving birth, black women who have an unplanned child are also significantly less likely than women who have not to be currently married, but are not less likely to have ever been married. Like black women, white women who have an unplanned teenage birth have significantly higher rates of poverty and welfare recipiency; they also have significantly lower family earnings and household income. PMID- 8405342 TI - Adolescent males' abortion attitudes: data from a national survey. AB - Roughly 13% of a nationally representative sample of 1,880 15-19-year-old males approve of abortion in each of eight circumstances presented to them, while about 4% disapprove in every instance. The proportions agreeing that abortion is acceptable range as high as 85-90% if the pregnancy endangers the woman's health or results from rape. Any type of religious affiliation, especially religious fundamentalism, is related to weaker support for abortion; an even stronger correlate of abortion attitudes is the importance of religion to the respondent. Abortion attitudes vary little by race after other social background factors are controlled. Those with more liberal attitudes toward premarital sex and those who perceive that they would be upset if they became a father in the immediate future are particularly likely to express acceptance of abortion. Roughly 61% of adolescent males do not feel that it would be all right for a woman to have an abortion if her partner objects, indicating a possible "gender conflict of interest" over the abortion issue. PMID- 8405343 TI - Multiple heterosexual partners and condom use among Hispanics and non-Hispanic whites. AB - A telephone survey of 1,592 Hispanic and 629 non-Hispanic white men and women aged 18-49, randomly selected from nine states in the northeastern and southwestern United States, found that married Hispanic men are more likely to have had two or more heterosexual partners in the previous 12 months than are married non-Hispanic men (18% and 9%, respectively). A large proportion of unmarried men (60% of Hispanics and 54% of non-Hispanic whites) report having had more than one partner in the past 12 months. After adjusting for other variables, the odds of having multiple partners are 2.5 times higher among Hispanic men aged 18-24 than among those aged 41-49 and 1.8 times higher among those who live in the Northeast than among those in the Southwest. Highly acculturated Hispanic men are less likely to have multiple partners than are less acculturated men. Among Hispanic women, those who are moderately or highly acculturated are more likely to have multiple partners (odds ratios of 4.9 and 8.4, respectively) than are less acculturated women. About half of men and women with multiple partners report always using condoms with secondary heterosexual partners. PMID- 8405344 TI - Women's contraceptive attitudes and use in 1992. AB - Women aged 15-44 rate the pill, the condom, vasectomy and female sterilization most highly, according to 1992 data from an annual survey by Ortho Pharmaceutical Corporation of contraceptive attitudes and method use. The 6,955 survey respondents underrepresent women who are black or who have household annual incomes greater than $50,000, but they are similar to all American women in age, marital status and region of the country. About 74-84% of women giving an opinion view these methods favorably and 64% rate the hormonal implant favorably. The proportion of unmarried women who had had intercourse increased from 76% in 1987 to 86% in 1992. As a result, proportions of women at risk of unintended pregnancy rose from 72% to 77%. Contraceptive use also rose, from 92% to 94%. The most commonly used method is the pill (39%), followed by the condom (25%), female sterilization (19%) and vasectomy (12%). Married women exposed to the risk of unintended pregnancy are more likely to use sterilization (48%), while unmarried women are more likely to use the pill (52%) and the condom (33%). Pill use has increased since 1987, especially among married women, and condom use has increased among all women. Among unmarried women at risk of unintended pregnancy, condom use rose from 18% in 1987 to 33% in 1992. Among condom users, 40% of unmarried users and 13% of married users also use another method. PMID- 8405345 TI - Project offers counseling and condoms to Philadelphia students. PMID- 8405346 TI - For speedy Medicaid enrollment, Indianapolis sends a 'Babygram'. PMID- 8405347 TI - Estimating the prevalence of homosexual behavior. PMID- 8405348 TI - Revisiting the notion of hierarchy. AB - The traditional definition of hierarchy in terms of power has rendered the image of the hierarchical therapist a fit subject for constructionist critiques. This essay uses an alternative definition of hierarchy, cast in temporal and developmental terms, as the basis for a revised understanding of the role of hierarchy in the therapeutic process. It is argued that when therapy is constructed as a "framing-developing hierarchy," rather than as a "power hierarchy," then constructionist aspirations to create a therapy that is nonjudgmental, nonpathologizing, and noncontrolling are realized. PMID- 8405349 TI - Power and hierarchy: let's talk about it! PMID- 8405350 TI - The therapist as authority figure. PMID- 8405351 TI - Hierarchy: the imbalance of risk. PMID- 8405352 TI - Disordered communication and grieving in deaf member families. AB - When a deaf child is born to hearing parents, a grieving process is initiated in the parents. Unresolved grieving over their child's deafness often makes it difficult for hearing parents to accept the importance of signing, thus increasing the child's problems--a further source of grief for these parents. Clinical illustrations are provided of (1) the reciprocal relationship between disruption of the mourning process and disturbance of communication between family members, and (2) the transmission of the dysfunctional relationship between hearing parents and deaf children to the subsequent relationship between the deaf children, when they reach adulthood, and their hearing children. PMID- 8405353 TI - Ecosystemic training: conjoining supervision and organizational development. AB - Training in systems therapy is itself a systems process that is influenced by the organizational context in which it occurs. Systems trainers need to apply their skills to the interface between them and the agency, otherwise they face the danger that their impact will be eroded by organizational difficulties in "internalizing" their approach. Furthermore, there is much to be gained by working in this domain, especially if the training stimulates and becomes intertwined with organizational development. This coevolutionary process then facilitates integration of the systems model by both the trainees and the agency. Three variables, important for working at this interface, are analyzed: the approach of the supervisor, the composition and dynamics of the training group, and the treatment philosophy and work patterns of the institution. The implementation of this approach is demonstrated through a 4-year project of training and program development with Israel's Youth Protection Authority. PMID- 8405354 TI - Competence in children at risk for psychopathology predicted from confirmatory and disconfirmatory family communication. AB - The relationship between confirmation/disconfirmation in parental and family communication and offspring social competence was examined in 59 families in which at least one of the parents had been hospitalized for a functional psychiatric disorder. Communication samples were obtained using the Consensus Rorschach procedure both with parental couples and with parent-child family units. The communication was analyzed using the Confirmation-Disconfirmation Coding System (CONDIS). The competence at school of 7-and 10-year-old boys was rated by both peers and teachers. Competence at home was rated by the parents. The results indicated that the more competent the high-risk children were, both at school and at home, the more their family communicated in confirmatory ways and the less they communicated in disconfirmatory ways. Furthermore, although the parental couple CONDIS score and the family CONDIS score were modestly correlated, each contributed separately to the prediction of offspring competence. These communication data were not significantly related to parental psychopathology, neither severity of parental impairment nor the diagnosis of the patient-parent. PMID- 8405355 TI - FACES III and the Kvebaek Family Sculpture Technique as measures of cohesion and closeness. AB - In a longitudinal study of psychosocial factors in children with recent onset of rheumatic disease (N = 72), it was hypothesized that FACES III cohesion sum scores would be correlated with family closeness as measured by the Kvebaek Family Sculpture Technique (KFST). A second aim was to examine how these instruments were associated with semi-structured interview assessments of child and family psychosocial function, and whether these associations were linear or curvilinear. FACES III cohesion sum scores and KFST family mean interpersonal distance were not correlated, whereas the cohesion sum scores were related to the mother-father distance on the KFST. Where associations with child and family psychosocial function were found, they were linear. The results support the usefulness of a distinction between cohesion as a family or group characteristic and dyadic closeness, especially between the parent figures. The need for further conceptual clarification and more thorough empirical validation procedures in this area is emphasized. PMID- 8405356 TI - Conceptualizing social support in families of children with special health needs. AB - The concept of social support is used increasingly to understand families and their functioning. While conceptualization of the support process is regrettably absent for much research on families, earlier models developed for examining social support of individuals can enlighten research on families. The history of the social support concept is presented along with an overview of current typologies of social support and models of how it impacts physical and mental health. Research on the social support of families with children with special needs is reviewed relative to these issues. Greater recognition of a comprehensive model of support is advocated. Recommendations are made for longitudinal research on temporal patterns of utilization and satisfaction with support, and for consideration of cultural contexts in interpreting social supports. PMID- 8405357 TI - DSM-IV and describing problems in family therapy. PMID- 8405358 TI - Relational diagnosis: an idea whose time has come? AB - Many marital and family therapists in the U.S., as well as the major organizations to which they belong, recognize that the DSMs and ICDs have become the accepted manuals for classification of individual mental and emotional dysfunctions. Because we/they have come to believe that many disorders emanate from interpersonal problems rather than from individual intrapsychic distress, a strong movement has mushroomed to develop a diagnostic system that describes relational disorders and to press for the inclusion of these in future versions of DSM and ICD. This commentary chronicles the evolution and accomplishments to date of those involved in this effort. PMID- 8405359 TI - cDNA cloning and expression of cysteine synthase B localized in chloroplasts of Spinacia oleracea. AB - The cDNA clones for cysteine synthase B, which is localized in chloroplasts of Spinacia oleracea L., were isolated by screening a library with synthetic oligonucleotides encoding a partial peptide sequence of the purified protein. Nucleotide sequence analysis revealed an open reading frame encoding a polypeptide of 383 amino acids containing a putative transit peptide of 52 amino acids. A bacterial expression vector of the cDNA clone could genetically complement an Escherichia coli auxotroph lacking cysteine synthase and could produce the functionally active and immuno-reactive cysteine synthase in E. coli. RNA blot hybridization suggested that the transcripts were primarily accumulated in leaves of spinach. PMID- 8405360 TI - Expression in E. coli and purification of a chimeric p22-NS3 recombinant antigen of hepatitis C virus (HCV). AB - A recombinant antigen (p22-NS3), possessing putative HCV nucleocapsid protein (p22) and non-structural protein 3 (NS3) epitopes, was heavily expressed in E. coli and purified. The p22-NS3 purified recombinant antigen strongly reacts with sera containing human antibodies directed against p22 and NS3 providing a starting point for the design of an HCV single all-encompassing antigen for a blood screening assay. PMID- 8405361 TI - The role of N-glycosylation in the targeting and stability of GLUT1 glucose transporter. AB - The cDNAs encoding the GLUT1 glucose transporter protein were altered by site directed mutagenesis at consensus sites for the addition of N-linked glycosylation. These cDNAs were transfected into CHO cells with an expression vector and the subcellular distribution and stability of the expressed glycosylation-defective GLUT1 protein were analyzed. Immunohistochemical analysis with a specific antibody demonstrated that a significant portion of glycosylation defective GLUT1 protein remained in the intracellular compartment. By contrast, most of the wild-type GLUT1 protein expressed with the same procedures resided in the plasma membranes. Metabolic labeling studies revealed that the half-life of the glycosylation-defective GLUT1 protein was significantly shorter than that of wild-type GLUT1 protein. These results indicate that N-glycosylation of the glucose transporter affects its intracellular targeting and protein stability. PMID- 8405362 TI - Reduced glutamate decarboxylase activity in rat islet beta cells which survived streptozotocin-induced cytotoxicity. AB - Rat pancreatic beta cells exhibit a 16-fold higher glutamate decarboxylase (GAD) activity than islet non-beta cells, but a similar glutamate dehydrogenase (GDH) activity. beta Cells which survive exposure to 2 mM streptozotocin only contain 10 percent of the GAD activity of control cells, but their GDH activity remains unaltered. Culture of streptozotocin-treated beta cell preparations with 2 mM nicotinamide reduces the number of dead cells and prevents in part the decline in GAD activity of surviving beta cells. These data indicate that loss in activity of the beta cell specific enzyme GAD can serve as marker for beta cells which survived a destructive process. It is furthermore demonstrated that nicotinamide increases the percent surviving cells and decreases their loss in GAD activity. PMID- 8405363 TI - Beta-crystallins insolubilized by calpain II in vitro contain cleavage sites similar to beta-crystallins insolubilized during cataract. AB - Incubation of soluble proteins from rat lens with the protease calpain II caused the precipitation of beta-crystallin polypeptides. Two-dimensional electrophoresis and sequence analysis identified beta-crystallin polypeptides both before and after their precipitation by calpain II. beta-crystallin polypeptides precipitated by calpain were cleaved at their NH2-terminal extensions. These cleavage sites were similar to cleavage sites occurring in beta crystallin polypeptides precipitated during formation of experimental cataract induced by an overdose of selenite. These data suggested that calpain II caused beta-crystallin insolubilization during cataract formation, and indicated that the process can be mimicked in vitro. PMID- 8405364 TI - On the origin of a sustained increase in cytosolic Ca2+ concentration after a toxic glutamate treatment of the nerve cell culture. AB - A sustained increase of cytosolic Ca2+ concentration, [Ca2+]i, (Ca2+ plateau) was induced by a 15-min treatment with 50 microM glutamate of cultured cerebellar granule cells and hippocampal neurons in a Mg(2+)-free solution. Plateau proved to be insensitive to inhibition of Na+o/Ca2+i exchange caused by removal external Na+ in the post-glutamate period. A approximately 10(5)-fold reduction of [Ca2+]o (from 1.5 mM to 20 nM) in the post-glutamate period caused in most cells only a slow and small decrease in [Ca2+]i, although the same low-Ca2+ trial before glutamate treatment caused in hippocampal cells very quick blockade of spontaneous [Ca2+]i oscillation and a decrease in the basal [Ca2+]i. The results suggest that the Ca2+ plateau is due to a suppression of the Ca2+ extrusion from the cell (in particular via Na+/Ca2+ exchange) rather than from a persistent increase in Ca2+ permeability of neuronal membrane. PMID- 8405365 TI - On the role of intracellular concentration of Ca2+ and H+ in thymocyte death after irradiation. AB - The role of intracellular Ca2+ and H+ concentrations in radiation-induced interphase death of rat thymocytes has been studied. In response to concanavalin A treatment in the Ca(2+)-containing medium, or to the CaCl2 treatment in the Ca(2+)-free medium, the [Ca2+]i rise in irradiated cells was as in the non treated cells. No changes in the level of [Ca2+]i and pHi were found within 1 h after irradiation of thymocytes with a dose of 6 Gy. 15 microM 5-(N-ethyl-N isopropyl)-amiloride, an inhibitor of Na+/H+ exchange, did not affect the DNA fragmentation. The fragmentation was prevented by 2-4 microM (1-[bis(4 chlorophenyl)methyl]-3-[2-(2,4-dichlorophenyl)]-2-[(2,4- dichlorophenyl)-methoxy] ethyl)-1-H-imidazolium chloride, an inhibitor of calmodulin. The above data indicate that triggering of interphase death in irradiated thymocytes is not mediated by changes in either [Ca2+]i or pHi. Such changes seem to be involved in intermediate steps of the interphase death process. PMID- 8405366 TI - Contraction of smooth muscle by activation of endothelin receptors on autonomic neurons. AB - Endothelin receptors, predominantly of the ETB type, were localized to cell bodies, processes, and varicosities of cholinergic and adrenergic intramural autonomic neurons that were present in primary cultures of guinea pig tracheal smooth muscle. Stimulation of the neuronal ETB receptor produced a tetrodotoxin sensitive increase in the intracellular calcium concentration in neurons which was followed by contraction of the neighboring smooth muscle cells. These observations suggest that endothelins can induce smooth muscle contraction by means of a neuronally mediated mechanism, in addition to their direct actions on the smooth muscle. PMID- 8405367 TI - Requirement of the calcium channel beta subunit for functional conformation. AB - The cardiac dihydropyridine-sensitive L-type calcium channel was stably expressed in Chinese hamster ovary cells by transfecting the rabbit cardiac calcium channel alpha 1 subunit cDNA with or without coexpression of the beta subunit of skeletal muscle calcium channel. Whereas coexpression of the beta subunit significantly increased DHP binding activity and calcium channel activity, it did not affect the amount of the alpha 1 subunit expressed, as judged by RNA blot hybridization analysis and immunoblotting analysis. The results suggest that association with the beta subunit is necessary for the alpha 1 subunit protein to take a proper conformation suitable for a functional calcium channel. PMID- 8405368 TI - An interaction of beta-amyloid with aluminium in vitro. AB - We have used circular dichroism spectroscopy to confirm that, in a membrane mimicking solvent, A beta P(1-40) adopts a partially helical conformation and we have demonstrated the loss of this structure in the presence of physiologically relevant concentrations of aluminium. This is the first evidence of a direct biochemical interaction between aluminium and beta-amyloid and may have important implications for the pathogenesis of Alzheimer's disease. PMID- 8405369 TI - The catalytic domain of human hepatitis delta virus RNA. A proton nuclear magnetic resonance study. AB - We have obtained and analyzed the 600 MHz proton NMR spectra of a 74-mer RNA derived from the catalytic domain of hepatitis delta virus genomic RNA (HDV RNA) to determine its secondary structure. Deconvolution of the NMR spectrum obtained at 32 degrees C indicates that part of the 74-mer RNA molecule may exist in multiple conformations in equilibrium. The major conformer contains two A-U base pairs and 14 +/- 2 G-C base pairs. It appears to contain no standard G-U base pairs. Our NMR melting study suggests that this conformer has at least two stem loop regions. One of the regions has been identified to be a tetra-loop. We have assigned five imino proton resonances of the tetra-loop stem. Our data is consistent with the pseudoknot model of Perrotta and Been. PMID- 8405370 TI - Interaction of mutagenic tryptophan pyrolysate with DNA. CD spectral study on the binding specificity. AB - The interactions of DNA duplexes with 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), a potent mutacarcinogen isolated from tryptophan pyrolysate, have been studied using CD spectroscopy. The results are that (a) the spectral change of B form DNA caused by the interaction with Trp-P-1 is biphasic, i.e. the enlargement of CD bands characteristic to the B-DNA conformation in the range of r ([Trp-P 1]/ DNA]) = 0-2.5, followed by the rapid transition to the non-B conformation at r > 2.5; (b) this transition degree of B to non-B conformation of DNA is not necessarily dependent on the G-C content; and (c) the salt-induced Z-DNA is transformed to B-DNA (0 < r < 0.1) and then to non-B-DNA (r > 5), depending on the concentration of Trp-P-1 added. These data indicate that the non-covalent interaction of Trp-P-1 with DNA is mainly dependent on the B-DNA conformation. PMID- 8405371 TI - Two-dimensional 1H NMR spectra of ferricytochrome c551 from Pseudomonas aeruginosa. AB - The full assignment of 1H NMR signals of heme proton resonances of ferricytochrome c551 from Pseudomonas aeruginosa has been performed by means of 2D NMR experiments. This technique allows the complete and unequivocal assignment of all heme resonances, including methylene resonances of the propionic groups, directly implicated in the pH dependence of the redox properties of cytochrome c551. PMID- 8405372 TI - Sunlight-induced mutagenicity of a common sunscreen ingredient. AB - We have tested the mutagenicity of a UV-B sunscreen ingredient called Padimate-O or octyl dimethyl PABA, which, chemically speaking, is identical to an industrial chemical that generates free radicals when illuminated. It is harmless in the dark but mutagenic in sunlight, attacking DNA directly. A commercial sunscreen containing Padimate-O behaves in the same way. UV-A in sunlight also excites Padimate-O, although less than UV-B. Some related compounds, including a known carcinogen, behave similarly. As mutagens may be carcinogenic, our results suggest that some sunscreens could, while preventing sunburn, contribute to sunlight-related cancers. PMID- 8405373 TI - Identification of a cross-linked double-peptide from the catalytic site of the Ca(2+)-ATPase of sarcoplasmic reticulum formed by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate. AB - The Ca(2+)-ATPase from sarcoplasmic reticulum can be inhibited by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate. The chemically and monofunctionally activated inhibitor introduces an intramolecular cross-link between two neighbouring peptides of the active site. This can be followed by the reduced mobility of the ATPase upon SDS-PAGE analysis which becomes even more pronounced after limited trypsinolysis. After cleavage of the cross-linked ATPase molecule by cyanogen bromide and separation of the peptides a double-peptide can be detected which upon sequencing can be identified as part of the phosphorylation and the nucleotide binding site, respectively. PMID- 8405374 TI - Evidence for three binding sites for C3 (hemolytically inactive), C3b and C3d on a CR2-positive Burkitt lymphoma-derived cell line (Raji). AB - The present investigation shows that C3 (hemolytic inactive) as well as C3b and C3d bind Raji, a CR2-positive Burkitt lymphoma-derived cell line. Pretreatment of the cells with OKB-7 inhibited the binding of C3, whereas pretreatment with HB-5 inhibited the binding of C3b. Furthermore, the cells coated either with OKB-7 or HB-5 bound high amounts of C3d. TPA-treated cells showed binding for C3b and weak binding for C3 and C3d. Taken together, the data suggest that Raji cells may express three binding sites for C3, C3b and C3d which can be differently modulated by anti-CR2 MoAbs and TPA. PMID- 8405375 TI - Participation of cathepsin B in processing of antigen presentation to MHC class II. AB - Cellular and humoral immune responses to vaccines of hepatitis B type and rabies were inhibited by specific inhibitors of cathepsin B, specific synthetic substrates of cathepsin B and anti-cathepsin B antibody. Therefore the lysosomal cathepsin B of antigen presenting cells plays an essential role in processing of these antigens for presentation to MHC class II. One of the active sites of cathepsin B, VN217-222 shares highly homologous sequences with a part of the desetope, a binding domain of antigenic peptides, VN57-62 of MHC class II, beta chain. This evidence suggests that the peptides processed by the substrate specificity of cathepsin B exhibit a common affinity to bind with the desetope of MHC class II, beta-chain. PMID- 8405376 TI - Amidating processing enzyme complex for bioactive peptides (PAM) shows differences in specific activity and form in secretory granules isolated from the proximal and distal parts of the hypothalamo-neurohypophyseal tract in rats. AB - In rats the PAM specific activity in hypothalamic and neurohypophyseal extracts was 0.58 +/- 0.8, respectively 1.78 +/- 0.6 nmol.mg prot.-1 x h-1 (n = 5). PHM specific activity in the soluble part of the granules was higher in the neurohypophyseal than in the hypothalamic granules, and the fraction of total PHM and PAL present in the soluble part increased with the distance from the hypothalamus from some 45% to approx. 85%. Western blots of membrane and soluble granule fractions showed prevalence of higher mol. wt. forms in hypothalamic granules. It would appear that higher mol. wt. forms of PAM are processed by proteolytic enzymes during transport in the neuron and that non-neural cells in the neurohypophysis have a considerable PAM activity. PMID- 8405377 TI - Characterization of NCAM diversity in cultured neurons. AB - A single transcript of the NCAM gene undergoes differential processing resulting in a multiplicity of mRNAs and their translation products. In this study, the diversity of NCAM in rat primary neuronal cultures was investigated utilizing immuno- and Northern blot analyses. NCAM polypeptides of 190 kDa (NCAM-A) and 135 kDa (NCAM-B) were shown to be associated with the neuronal phenotype. These data were confirmed by Northern blotting, which in both neocortical neurons and cerebellar granule neurons revealed mRNA classes of 7.4 kb and 6.7 kb encoding for NCAM-A and -B, respectively. However, oligonucleotide probes, specific for selected exons or exon combinations, revealed special features of cerebellar granule neurons as compared to neocortical neurons: expression of 4.3 kb NCAM mRNA, a relatively low amount of VASE-containing variants, and an apparent lack of mRNA species containing exons alpha and an AAG insert between exons 12 and 13. Distinct patterns of NCAM mRNA may putatively be related to the regional origin and functional specificity of the investigated neurons. PMID- 8405378 TI - Detection of a 10 kDa breakdown product containing the C-terminus of the D1 protein in photoinhibited wheat leaves suggests an acceptor side mechanism. AB - Photoinhibition of intact leaves of wheat generates a 10 kDa breakdown product which is clearly observed both at 4 degrees C and 25 degrees C. Selective immunoblotting has shown that the 10 kDa fragment contains the C-terminus of the D1-protein and, under the conditions employed, supports an acceptor side mechanism for photoinhibition in vivo. Although a corresponding 23 kDa N-terminal D1-fragment was not detected our results are consistent with the argument that the primary cleavage site is in the loop joining putative transmembrane segments IV and V. PMID- 8405379 TI - Mos is degraded by the 26S proteasome in a ubiquitin-dependent fashion. AB - Mos, the c-mos proto-oncogene product, is a key regulator of cell cycle progression. Recently, rapid turnover of Mos in an early stage of meiotic maturation of Xenopus oocytes was found to be mediated by the ubiquitin pathway, but the protease responsible for its breakdown was not identified. In the present study, we found that 35S-labeled Mos synthesized in an in vitro transcription/translation system was degraded ATP- and time-dependently by the 26S proteasome, but not by the 20S proteasome, in the presence of a ubiquitin ligation system. The 26S proteasome did not degrade a mutant Mos in which Ser3 was replaced by Asp3 that is metabolically stable in oocytes, indicating a similarity in the proteolytic events in vivo to those observed in vitro in the present work. This is the first demonstration that the proteasome catalyzes the ATP-dependent degradation of a naturally occurring, short-lived oncoprotein by the ubiquitin pathway. This finding suggests that the proteasome may regulate the intracellular stability of various oncoproteins. PMID- 8405380 TI - Receptor-binding capability of pancreatic phospholipase A2 is separable from its enzymatic activity. AB - Mammalian pancreatic phospholipase A2 (PLA2-I) has its specific receptor through which PLA2-I induces a variety of biological responses. In this study, a fundamental relationship between the enzymatic and the receptor-binding activities of PLA2-I was investigated. The specific binding of PLA2-I to the receptor was found to be independent of Ca2+ which is requisite for the PLA2 activity. On the basis of this observation, we designed and produced mutant PLA2 Is without Ca(2+)-binding abilities in order to demonstrate that the structural requirement for the enzymatic activity of PLA2-I is not identical with that for its receptor-binding reaction. These mutant PLA2-Is lost almost all enzymatic activity through a disturbance at the Ca(2+)-binding site, as expected, but still retained a substantial affinity to the receptor, allowing us to conclude that the receptor-binding reaction of PLA2-I is separable from its catalytic action. PMID- 8405381 TI - Production of monoclonal antibodies that inhibit ADP-ribosylation of small GTP binding proteins catalyzed by Clostridium botulinum ADP-ribosyltransferase C3. AB - Four monoclonal antibodies that inhibited ADP-ribosylation of 23 kDa protein(s) of ascidian eggs catalyzed by Clostridium botulinum ADP-ribosyltransferase C3 were produced. They also inhibited C3-catalyzed ADP-ribosylation of the 24 kDa protein of rat liver cytosol. By the immunoprecipitation technique, it was found that they recognized small GTP-binding proteins of ascidian eggs and mammalian brains, but did not interact with the rat brain activator of the ADP ribosyltransferase reaction. The antibody can also immunoprecipitate recombinant Rho A irrespective as to whether the Rho A is the GDP-bound form or the GTPrS bound form. Thus the antibodies are novel and useful tools in analyzing the physiological roles of the Rho family of GTP-binding proteins. PMID- 8405382 TI - Glucuronidation of thyroid hormone by human bilirubin and phenol UDP glucuronyltransferase isoenzymes. AB - The glucuronidation of thyroid hormone by UDP-glucuronyltransferases (UGTs) stably transfected in Chinese hamster V79 lung fibroblasts was investigated. Human bilirubin UGT (HP3) and phenol UGT (HP4) both catalysed the glucuronidation of T4 and rT3, whereas glucuronidation of T3 was not significant, rT3 was the preferred substrate for both isoenzymes, glucuronidation rates being 1.6- and 6.4 times higher than conjugation of T4 by HP3 and HP4 clones, respectively. This is the first identification of thyroid hormone as potential alternative endogenous substrate for bilirubin UGT. PMID- 8405383 TI - The KH domain occurs in a diverse set of RNA-binding proteins that include the antiterminator NusA and is probably involved in binding to nucleic acid. AB - New findings are presented for the approximately 50 residue KH motif, a domain recently discovered in RNA-binding proteins. The conserved sequence is approximately 10 residues larger than previously reported. Profile searches have revealed new members of this family, including two, E. coli NusA and human GAP associated p62 phosphoprotein, for which RNA-binding data exists. A nusA homolog was detected in the RNA polymerase gene complex of six archaebacterial species and may encode an antiterminator. All KH-containing proteins are linked with RNA and the KH motif most probably functions as a nucleic acid binding domain. PMID- 8405384 TI - Identification of putative internalization signals in prion proteins. AB - Prion proteins were found to contain regions in their cytoplasmic domains that have significant structural homologies to molecular trafficking signals for internalization of membrane proteins. These regions may facilitate the endocytosis of prion proteins, which appears to be a first step in their conversion to the abnormal, amyloidogenic form. PMID- 8405385 TI - Penicillin binding protein 2x as a major contributor to intrinsic beta-lactam resistance of Streptococcus pneumoniae. AB - The production and purification to protein homogeneity of a soluble form of PBP2x from a cefotaxime-resistant Streptococcus pneumoniae strain is reported. It was obtained by a site-directed deletion of the membrane anchor in the corresponding gene, a method similar to that successfully utilized for the production of PBP2x from a cefotaxime-sensitive wild type strain. The kinetic parameters characterizing the interactions of both cefotaxime-resistant and -sensitive proteins have been determined and compared. The results are in agreement with the identification of PBP2x as the primary target for cefotaxime in the sensitive strain and as probably one of several targets in the resistant strain. PMID- 8405386 TI - Biosynthesis of vitamin B12. Discovery of the enzymes for oxidative ring contraction and insertion of the fourth methyl group. AB - In the vitamin B12 biosynthetic pathway the enzymes responsible for the conversion of precorrin-3 to precorrin-4 have been identified as the gene products of cobG and cobJ from Pseudomonas denitrificans. CobG catalyzes the oxidation of precorrin-3 to precorrin-3x (a hydroxy lactone) whereas CobJ is a SAM-dependent C-17 methyl transferase and is necessary for ring contraction. A mechanism for ring contraction is proposed. PMID- 8405387 TI - Essential role of the Arg112 residue of cytochrome P450cam for electron transfer from reduced putidaredoxin. AB - Cytochrome P450cam (CYP101) of Pseudomonas putida PpG1 in which Arg112 is substituted by Cys was isolated by in vitro random mutagenesis of the camC gene DNA coding for P450cam. The absorption spectra of the purified mutant enzyme were similar to those of the wild type enzyme, but its substrate-dependent NADH oxidation activity in the presence of putidaredoxin (Pd) and putidaredoxin reductase (PdR) was extremely low. The rate constant of electron transfer from reduced Pd to the heme of the mutant P450cam, measured on an anaerobic stopped flow apparatus, was 1/400 of that of the wild type enzyme and the dissociation constant of the mutant P450cam for oxidized Pd was several fold higher than that of the wild type enzyme. A considerable decrease in mid-point potential of the mutant enzyme was also noted. We conclude that Arg112, which is located on the surface of the P450cam molecule and hydrogen-bonded to one of the heme propionate chains, plays an essential role in the electron transfer from Pd. PMID- 8405388 TI - Bovine inositol monophosphatase. Studies on the binding interactions with magnesium, lithium and phosphate ions. AB - Rapid equilibrium dialysis has been used to show that recombinant bovine brain inositol monophosphatase binds one equivalent of Pi per subunit of enzyme. Pi is only bound in the presence of Mg2+ ions. The dissociation constant for the equilibrium is approximately 50 microM. This value of Kd is independent of the concentration of the Mg2+ ions and of the presence or absence of Li+ ions. Lithium ions which inhibit the enzyme uncompetitively are not able to support the binding of the Pi to the enzyme. The observation that Pi only binds in the presence of Mg2+ ions supports similar conclusions made in experiments which studied the protection of the enzyme from proteolytic degradation and chemical modification. PMID- 8405389 TI - Chronic ethanol administration enhances retinoic acid and triiodothyronine receptor expression in mouse liver. AB - Chronic alcoholism induces perturbations of storage and metabolization of retinol and related compounds. After 6 months of ethanol consumption we have observed in mouse liver an increased expression of Tri-iodothyronine receptors (TR) while the expression of retinoic acid (RA) receptors (RAR) was unaffected. After 10 months of alcoholization the TR expression was strongly increased and the RAR expression was also increased. At this time the activity of aldehyde dehydrogenase and that of alcohol dehydrogenase, two enzymes involved in biosynthesis of RA from retinol, were similar in the liver of alcoholized and pair-fed mice. Thus it can be hypothesized that (i) the change of RAR expression was, at least in part, the result of a change of TR expression (result in agreement with previous data), (ii) the increased expression of RAR could induce apoptosis and subsequently liver necrosis. PMID- 8405390 TI - The crystal structure of triacylglycerol lipase from Pseudomonas glumae reveals a partially redundant catalytic aspartate. AB - The family of lipases (triacylglycerol-acyl-hydrolases EC 3.1.1.3) constitutes an interesting class of enzymes because of their ability to interact with lipid water interfaces, their wide range of substrate specificities, and their potential industrial applications. Here we report the first crystal structure of a bacterial lipase, from Pseudomonas glumae. The structure is formed from three domains, the largest of which contains a subset of the alpha/beta hydrolase fold and a calcium site. Asp263, the acidic residue in the catalytic triad, has previously been mutated into an alanine with only a modest reduction in activity. PMID- 8405391 TI - Astacins, serralysins, snake venom and matrix metalloproteinases exhibit identical zinc-binding environments (HEXXHXXGXXH and Met-turn) and topologies and should be grouped into a common family, the 'metzincins'. AB - The X-ray crystal structures of two zinc endopeptidases, astacin from crayfish, and adamalysin II from snake venom, reveal a strong overall topological equivalence and virtually identical extended HEXXHXXGXXH zinc-binding segments, but in addition a methionine-containing turn of similar conformation (the 'Met turn'), which forms a hydrophobic basis for the zinc ion and the three liganding histidine residues. These two features are also present in a similar arrangement in the matrix metalloproteinases (matrixins) and in the large bacterial Serratia proteinase-like peptidases (serralysins). We suggest that these four proteinases represent members of distinct subfamilies which can be grouped together in a family, for which we propose the designation, metzincins. PMID- 8405392 TI - Repression of histone H5 gene expression in chicken mature erythrocytes is correlated with reduced DNA-binding activities of transcription factors Sp1 and GATA-1. AB - During the final stages of erythroid maturation, the expression of the chicken histone H5 gene ceases. The histone H5 promoter has binding sites for Sp1 and UPE binding protein. The 3' histone H5 enhancer has binding sites for Sp1, GATA-1 and NF1. Here, we show that the DNA-binding activities of transcription factors Sp1 and GATA-1 is reduced 5- to 10-fold in mature cells, while the activities of UPE binding protein and NF1 remain the same in mature and immature erythrocytes. The reduced activities of Sp1 and GATA-1 may contribute to the inactivation of the histone H5 gene in mature erythrocytes. PMID- 8405393 TI - Similarity between serine hydroxymethyltransferase and other pyridoxal phosphate dependent enzymes. AB - A structural homology of the pyridoxal-5'-phosphate (PLP)-dependent enzyme serine hydroxymethyltransferase (SHMT) with aspartate aminotransferase (AAT) is proposed. Although the two sequences are very dissimilar, a reasonable alignment was obtained using the profile analysis method. Sequences of AAT and dialkylglycine decarboxylase (DGD), for which crystal structure data are available, have been aligned on the basis of their structure superposition. A profile was then calculated and SHMT sequence aligned to it. Three of the four residues conserved in all aminotransferases (including the PLP-binding lysine) are matched. A profile search with DGD-AAT-SHMT profile is more selective and sensitive than individual sequence profiles for PLP-dependent enzyme detection. Potential homologies with the eryC1 gene product involved in erythromycin biosynthesis and with amino acid decarboxylases were observed. Homology with AAT will be used as a guideline for planning site-directed mutagenesis experiments on SHMT. PMID- 8405394 TI - Calcitonin gene expression in normal human liver. AB - Immunoreactive calcitonin (CT) is present in liver. This could represent hormone synthesized by liver cells, degraded or bound to specific receptors reported in this organ. We report here that the calcitonin gene is expressed in liver. We proved this by demonstrating, by PCR amplification using specific primers, the presence of calcitonin messenger in human liver and in primary cultures of human hepatocytes and detected by radioimmunoassay CT in hepatic tissues and cells. The synthesis of hormone by liver that also possesses specific receptors for CT favors the presence of an autocrine or paracrine system involving calcitonin in this organ. PMID- 8405395 TI - ras oncogene-induced transformation of a rat seminal vesicle epithelial cell line produces a marked increase of adenylate cyclase and protein kinase C activities. AB - Cells transformed by Kirsten murine sarcoma virus (Ki-MSV) have basal adenylate cyclase activity (AC) higher than control cells and comparable level of forskolin stimulated AC activity. Moreover, a higher protein kinase C (PKC) activity was found to be present in the transformed cells. The molecular mechanism underlying the increase of AC activity was investigated. Our findings strongly suggest that this biochemical event is due to a marked decrease of the alpha i negative control of the enzyme, even though the alpha i of transformed cells appears to possess fully functional domains interacting with both the effector enzyme and the agonist-activated receptor. PMID- 8405396 TI - Different target-site specificities of the hairpin ribozyme in cis and trans cleavages. AB - The hairpin ribozyme cleaves a phosphodiester bond at the 5' side of a 5'GUC3' sequence of an RNA with high efficiency. An RNA having a 5'GUA3' sequence instead of the GUC sequence is a poor substrate for this ribozyme. Here, we show that this is indeed so in a trans-acting ribozyme system, but in a cis-acting ribozyme system this ribozyme cleaves the 5' side of a GUA sequence as efficiently as the wild-type cleaves the GUC sequence. One base substitution in the ribozyme also affected the target-site specificity in the cis-acting system. PMID- 8405397 TI - Assembly of eukaryotic class III (N-out, C-in) membrane proteins into the Escherichia coli cytoplasmic membrane. AB - Class III membrane proteins lack cleavable signal peptides but adopt an N-out, C in topology with respect to their native membranes. We have analysed the fate of two eukaryotic class III plasma membrane proteins, human erythrocyte glycophorin C and influenza A virus M2 protein, in Escherichia coli. The N-terminal domains of both proteins were efficiently localised to the extracytoplasmic side of the bacterial cytoplasmic membrane. When beta-lactamase was fused to the C-terminus of glycophorin C it was localised to the cytoplasm, and protease treatment of spheroplasts caused a reduction in size of the fusion protein consistent with glycophorin C adopting its native topology in E. coli. PMID- 8405398 TI - Vitamin B6 deficiency causes activation of RNA polymerase and general enhancement of gene expression in rat liver. AB - The effect of vitamin B6 deficiency on the activity of RNA polymerase and expression of several mRNAs in rat liver was investigated. The activities of RNA polymerase I and II in the liver of vitamin B6-deficient rats were found to be higher than the control rats by 30%. The expression of several mRNAs, including mRNAs for beta-actin and glyceraldehyde-3-phosphate dehydrogenase, and the content of poly(A)+ RNA were also increased in vitamin deficiency. These observations suggest that vitamin B6 influences gene expression in the liver, at least in part, by modulating the activity of RNA polymerase. PMID- 8405399 TI - Identification of the barstar binding site of barnase by NMR spectroscopy and hydrogen-deuterium exchange. AB - The extracellular ribonuclease from Bacillus amyloliquifaciens, barnase, forms a tightly-bound one-to-one complex with its intracellular inhibitor barstar. The barstar binding site on barnase was characterized by comparing the differences in the chemical shift and hydrogen-deuterium exchange rates between free and bound barnase. Chemical shift assignments of barnase in the complex with barstar were determined from 3D NOESY-HMQC and TOCSY-HMQC spectra of a complex that had been prepared with uniformly 15N-labelled barnase and unlabelled barstar. Hydrogen exchange rates were obtained from an analysis of a series of [15N]HMQC spectra of a sample prepared in the same manner exchanged into D2O. The largest changes in either chemical shift or hydrogen-deuterium exchange rate are observed for residues located in the active-site and substrate binding loops indicating that barstar inhibits barnase activity by sterically blocking the active site. PMID- 8405400 TI - Selective inhibition of Colorado potato beetle cathepsin H by oryzacystatins I and II. AB - The use of oryzacystatins I and II, two cysteine proteinase inhibitors naturally produced in rice grains, represents an attractive way for the control of Coleoptera insect pests. The present study was done to analyze the inhibitory effect of recombinant oryzacystatins produced in Escherichia coli as fusion proteins against digestive proteinases of the major pest Colorado potato beetle (Leptinotarsa decemlineata Say). Both inhibitors had a significant effect on total proteolytic activity, but maximal inhibitions ranged from 20 to 80% for pHs varying from 5.0 to 7.0, respectively. This pH-dependent efficiency of plant cystatins was due to the selective inactivation of potato beetle cathepsin H, as demonstrated by the use of inhibitors with different specificities against cathepsins B and H. These results demonstrate the importance of having an adequate knowledge of insect proteinases specifically recognized by the inhibitors to be used in pest control strategies. PMID- 8405401 TI - Identification of 1,4-dihydropyridine binding domains within the primary structure of the alpha 1 subunit of the skeletal muscle L-type calcium channel. AB - Calcium channel blockers are drugs that bind to the alpha 1 subunit of L-type calcium channels and selectively inhibit ion movements through these channels. Determination of the mechanism of channel blockade requires localization of drug binding sites within the primary structure of the receptor. In this study the 1,4 dihydropyridine-binding site of the membrane bound receptor has been identified. The covalently labeled receptor was purified and digested with trypsin. The labeled peptide fragments were immunoprecipitated with sequence-directed antibodies. The data indicate the existence of at least three distinct dihydropyridine-binding domains within the primary structure of the alpha 1 subunit. PMID- 8405402 TI - Cloning, characterization and overexpression of a Streptococcus pyogenes gene encoding a new type of mitogenic factor. AB - A new type of mitogenic factor, termed MF, has been found in the culture supernatant of Streptococcus pyogenes and its N-terminal amino acid sequence has been determined. On the basis of this sequence, an S. pyogenes gene encoding MF was cloned and its nucleotide sequence was determined. The MF gene includes a long, open reading frame with 813 nucleotides capable of encoding the MF precursor protein with 271 amino acids. Removal of the putative 43 residues as a signal peptide results in the mature MF protein with 228 amino acids. The molecular mass of the mature MF is calculated as 25,363 which is consistent with the previously determined value of 25,370 for MF secreted from S. pyogenes. Neither nucleotide nor amino acid sequence homology was found between the mature MF and other streptococcal pyrogenic exotoxins, such as SPE A, SPE B and SPE C. The mature MF was recombinantly overexpressed as a fusion protein with glutathione S-transferase in Escherichia coli. The recombinant protein showed mitogenic activity in rabbit peripheral blood lymphocytes and immunoreactivity with the rabbit antiserum raised against the secreted MF from S. pyogenes. These data indicate that a unique gene encoding MF was cloned from S. pyogenes. PMID- 8405403 TI - Nucleotide and negatively charged lipid-dependent vesicle aggregation caused by SecA. Evidence that SecA contains two lipid-binding sites. AB - SecA which is an overall acidic protein was found to induce an increase in the turbidity of a solution of vesicles consisting of negatively charged phospholipids. This increase was found to be due to an aggregation of the vesicles mediated by SecA. The SecA-mediated vesicle aggregation was not found for zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphocholine and showed a large dependence on both temperature and ionic strength. Furthermore it was shown that ATP and to a lesser extent ADP+Pi were able to reduce the SecA-mediated vesicle aggregation, while no effect could be seen for a non-hydrolysable ATP analog AMP PNP. Using the steady state fluorescence anisotropy of 1,6-diphenyl-1,3,5 hexatriene present in 1,2-dioleoyl-sn-glycero-3-phosphoglycerol vesicles we could show that SecA inserts in the bilayer. Monolayer studies confirmed that SecA is able to cause close contact between two membranes and gave a direct insight into the different types of lipid-protein interactions involved. From our results we propose that the SecA monomer possesses two lipid-binding sites which in the functional dimer conformation are responsible for the SecA-mediated vesicle aggregation. PMID- 8405404 TI - PCR-based assignment of two myosin heavy chain cDNA clones to biochemically and histochemically defined single type IIB and IID fibers of rabbit muscle. AB - The present study assigns two as yet unidentified fast myosin cDNA clones to specific myosin heavy chain (MHC) isoforms and their mRNAs in different fiber types of rabbit skeletal muscle. Specific oligonucleotide primers were used for reverse transcription and polymerase chain reaction (PCR) to yield products of defined lengths. The method was sensitive enough to detect specific mRNA sequences in total RNA extracts from microdissected, freeze-dried, single-fiber fragments down to 16 ng dry weight. The fibers were typed histochemically and biochemically by their electrophoretically assessed MHC complement. The following results were obtained: clone pMHC20-40 was assigned to type IIB fibers and clone pMHC24-79 to type IID fibers. PMID- 8405405 TI - pH dependence of light-induced proton release by bacteriorhodopsin. AB - We have measured the current generated by light-activated proton release from bacteriorhodopsin into solution as a function of both pH and ionic strength. We find that proton release into solution decreases with increasing pH with an intrinsic pKa of 8.2 +/- 0.2. This pH dependence indicates that the deprotonation of a certain group inhibits or abolishes proton release. Under physiological conditions, this group either releases a proton directly into solution or interacts with the site of proton release. The most immediate candidates for this protonatable species are tyrosine-57, tyrosine-185, arginine-82, and water; acting individually or cooperatively. The salt dependence of the apparent pKa of this group also allows us to calculate the surface charge density of about -5 charges per bacteriorhodopsin, compatible with previous estimates. PMID- 8405406 TI - Novel oxaloacetate effect on mitochondrial Ca2+ movement. AB - Mitochondrial Ca2+ movement was investigated in the presence of oxaloacetate, which is widely known as a 'Ca(2+)-releasing' agent [1978, Proc. Natl. Acad. Sci. USA 75, 1690-1694]. It is demonstrated that rat liver mitochondrial are capable of net Ca2+ accumulation from the oxaloacetate supplemented assay mixture. Both the membrane energization and the cation uniport at the expense of oxaloacetate are shown to be specifically blocked by either arsenite or ammonium chloride. With respiratory inhibitors present, ADP is shown to be a prerequisite for a high Ca2+ capacity, which can be alternatively enlarged with a concomitant loss of the arsenite effect by an addition of an NADP(+)-specific reductant (isocitrate). Arsenite-sensitive production of NADPH is observed, thus suggesting coupling between pyridine nucleotide transhydrogenation and the cation uniport in mitochondria. The role of such a coupling mechanism in the uniporter-mediated Ca2+ fluxes in mitochondria is discussed. PMID- 8405407 TI - A G-protein-coupled 130 kDa phospholipase C isozyme, PLC-beta 4, from the particulate fraction of bovine cerebellum. AB - A 130 kDa PLC isozyme was purified from the particulate fraction of bovine cerebellum. This PLC was recognized by a polyclonal antiserum generated against the purified 97 kDa PLC-beta 4. Reconstitution of the purified 130 kDa PLC with the membranes of C6 Bu-1 cells in the presence of GTP gamma S or AlF4- resulted in PLC activation as well as the association of PLC with the membranes. Both the association and activation were revoked when the membrane was washed with 2 M KCl. The 97 kDa PLC-beta 4 did not associate with membranes. These data suggest that the 130 kDa PLC is the intact form of PLC-beta 4 the activity of which is likely to be regulated by a G-protein on the membrane. PMID- 8405408 TI - Widespread distribution of free D-aspartate in rat periphery. AB - We have identified and quantified free D-aspartate in adult rat peripheral organs using gas chromatographic-mass spectrometric and high-performance liquid chromatographic techniques. The level of free D-aspartate was highest in the adrenal, testis, spleen and pituitary, followed by the thymus, lung, ovary, placenta, pancreas and heart, and below the detection limit in the kidney, liver, brain, muscle and serum. These data provide the first evidence that a high level of free D-aspartate widely occurs in the adult rat periphery and suggest that the D-amino acid may be an endogenous substrate for D-aspartate oxidase. PMID- 8405409 TI - Replacement of His23 by Cys in a zinc finger of HIV-1 NCp7 led to a change in 1H NMR-derived 3D structure and to a loss of biological activity. AB - The nucleocapsid protein NCp7 of human immunodeficiency virus type 1 (HIV-1), which is necessary for the formation of infectious virions, contains two zinc fingers of the Cys-X2-Cys-X4-His-X4-Cys form. To elucidate the importance of this particular motif, well conserved in retroviruses and retroelements, we substituted the histidine residue by a cysteine in the first zinc binding domain 13VKCFNCGKEGHTARNCRA30. The structures of the mutated and native zinc complexed peptides were studied by two-dimensional 600 MHz 1H nuclear magnetic resonance (NMR) in aqueous solution. The nuclear Overhauser effects were used as constraints to determine the solution structures using DIANA software followed by AMBER energy refinement. The results show that native and mutant peptides fold into non-identical three-dimensional structures, probably accounting for the loss of retrovirus infectivity following the His-Cys point mutation. PMID- 8405410 TI - Expression, purification and crystallization of fully active, glycosylated human interleukin-5. AB - Recombinant human interleukin-5 (hIL-5) has been expressed at high levels and produced in large quantities in baculovirus infected Sf9 insect cells. The glycosylated protein was purified using immuno-affinity chromatography and gel filtration. Purified hIL-5 has been crystallized using standard vapour diffusion techniques with PEG as a coprecipitant. The crystals belong to the C2 space group and diffract to 2 A. PMID- 8405411 TI - Distribution and second messenger coupling of four somatostatin receptor subtypes expressed in brain. AB - The mRNA distribution in the brain and the coupling to cellular effector systems of four somatostatin receptors (SSTR1-4) was studied. All four SRIF receptor subtypes were expressed in cortex and hippocampus. In addition, SSTR1 mRNA was relatively abundant in the spinal cord whereas SSTR2 mRNA was also present in the striatum. The SSTR3 gene was predominantly expressed in the olfactory bulb and in the cerebellum. Conflicting results about the effector coupling of SSTR1-3 have been published previously. We have stably expressed human SSTR1-4 in HEK 293 human embryonal kidney cells. Agonist binding to the receptor subtypes, including the recently cloned SSTR4, inhibited the formation of forskolin-induced cAMP. Is is concluded that, in an appropriate cellular environment, all four receptor subtypes can functionally couple to the inhibition of adenylyl cyclase. PMID- 8405412 TI - cDNA deduced procionin. Structure and expression in protochordates resemble that of procholecystokinin in mammals. AB - Using an improved 3' RACE (PCR) amplification system containing oligonucleotide primer with an inosine at ambiguous codon positions and inverse PCR to amplify the 5' ends, we have isolated and characterized cDNA clones which encode cionin, a protochordean homologue of the mammalian hormones, cholecystokinin (CCK) and gastrin. The full-length cloned cDNA of 510 bp encoded a 128 amino acid preprocionin. Reverse transcription-PCR and subsequent cDNA cloning revealed that cionin mRNA is expressed in both the neuronal ganglion and the gut of the protochordate Ciona intestinalis. The primary structure of procionin resembles that of proCCK more than that of progastrin. Sequence-specific immunochemical analysis showed that the cionin gene is expressed also at peptide level in both the gut and the neural ganglion. The neuronal processing of procionin is, however, more complete both with respect to carboxyamidation and tyrosine O sulfation. Hence, the tissue-specific expression of the cionin gene in Ciona intestinalis resembles that of the CCK gene in mammals. PMID- 8405413 TI - Induction of stathmin expression during liver regeneration. AB - Stathmin is a 19 kDa cytoplasmic phosphoprotein proposed to act as a relay for signals activating diverse intracellular regulatory pathways. After two-thirds partial hepatectomy, the concentration of stathmin reached a peak between 48 and 72 hours, comparable to the levels observed in neonatal liver, at about 10 times the basal adult level. Stathmin then decreased to basal levels within 7 days, more rapidly than during postnatal tissue development (7 weeks), with no detectable change in its phosphorylation state. Interestingly, the mRNA for stathmin reached a peak much earlier than the protein, at 24 hours posthepatectomy, and decreased to a still detectable level until 96 hours after hepatectomy. Altogether, the present results further support the generatility of the implication of stathmin in regulatory pathways of cell proliferation and differentation during normal tissue development and posttraumatic regeneration. PMID- 8405414 TI - Selective proteolysis of the wheat Em polypeptide. Identification of an endopeptidase activity in germinating wheat embryos. AB - The 'Em' polypeptide is the most abundant cytosolic polypeptide in mature wheat embryos. It is selectively and completely degraded within the first 24 h of germination. Extracts from germinated embryos contain endopeptidase activities which degrade the Em polypeptide. These are separable into a major and minor component by ion-exchange chromatography and the use of inhibitors shows the major component to be a cysteine proteinase. This activity shows a strong preference for the Em polypeptide as a substrate, being inactive against polypeptides which are not developmentally regulated and showing only low activity towards developmentally related, but otherwise nonhomologous 'dehydrin' polypeptides. PMID- 8405415 TI - Purification of an endoproteinase that digests the wheat 'Em' protein in vitro, and determination of its cleavage sites. AB - Germinating wheat embryos contain two endoproteolytic activities which digest the prominent 'Em' polypeptide. These are easily assayed in clarified embryonic homogenates and are distinguishable by the pattern of their peptide products and by their different pH optima. One activity has a pH optimum of 4.0; the second activity is a cysteine endoproteinase with a preference for the 'Em' protein as its substrate. It is maximally active between pH 5.5 and 6 at 25 degrees C. Analysis of the early cleavage products of the cysteine proteinase indicates scissile bonds between residues Glu32-Ala33 and Asn36-Leu37 in the 'Em' polypeptide. This endoproteinase has been purified and identified as a single polypeptide species of ca. 38,000 kDa. PMID- 8405416 TI - Physical map of the extremely thermophilic bacterium Thermus thermophilus HB27 chromosome. AB - The physical map of the chromosome of Thermus thermophilus HB27 was constructed using three restriction enzymes; EcoRI, SspI and MunI by applying pulsed-field gel electrophoresis techniques. Although the genome size of 1.82 Mb was almost the same as that (1.74 Mb) reported for T. thermophilus HB8 [Borges, K.M., and P.L. Bergquist. (1993) J. Bacteriol. 175, 103-110], the MunI cleavage maps were different. A 240 kb plasmid was detected in HB27, and its physical map was also constructed. In addition, several genes were located on the chromosomal physical map. PMID- 8405417 TI - Cloning and sequence of the cDNA coding for rat type II Fc gamma receptor of mast cells. AB - A clone encoding the rat type II Fc gamma receptor was isolated from a cDNA library of the rat mucosal mast cell line RBL-2H3 and its sequence determined. The predicted amino acid sequence is highly homologous with the mouse type II as well as rat type III Fc gamma receptors. The site of alternative splicing which generates the mouse Fc gamma Rb2 isoform is completely conserved. Hence we consider the new sequence to encode a rat Fc gamma RII b2 isoform. In further analogy to the mouse receptor, no consensus motif known to be involved in accessory chain association was observed in the transmembranal domain. The importance of the identification of this receptor for investigation of the immunological stimulation of mast cells is discussed. PMID- 8405418 TI - Isolation and amino acid sequence of the 30S ribosomal protein S19 from Mycobacterium bovis BCG. AB - The 30S ribosomal proteins from Mycobacterium bovis BCG were separated by reverse phase-high performance liquid chromatography (RP-HPLC). The isolated proteins were analyzed by SDS-PAGE, blotted on PVDF-membranes and subjected to sequence analyses using a gas-phase sequencer to correlate them to those of the well studied Escherichia coli and Bacillus stearothermophilus ribosomes. Moreover, the internal amino acid sequence of one ribosomal protein, MboS19, which is homologous to E. coli ribosomal protein S19 (EcoS19) and B. stearothermophilus ribosomal protein S19 (BstS19), was further analyzed by sequencing its internal peptides and two segments from the N- and C-termini of the protein were selected to deduce the sequence of two oligonucleotide primers which were used in a polymerase chain reaction. Using the amplified DNA fragment thus obtained as a hybridization probe, the gene encoding protein S19 was identified and cloned. Sequence analysis of the DNA fragment, together with peptide sequence analysis could determine the complete amino acid sequence of MboS19. This sequence proved to be 64% and 71% identical to those of the corresponding S19 proteins from the eubacteria E. coli, and B. stearothermophilus, respectively; 33% of the residues of MboS19 were identical to those in the archaebacterial ribosomal protein HmaS19. PMID- 8405419 TI - Carboxy-terminal processing of the large subunit of [NiFe] hydrogenases. AB - Two electrophoretic forms of the large subunit of the soluble periplasmic [NiFe] hydrogenase from Desulfovibrio gigas have been detected by Western analysis. The faster moving form co-migrates with the large subunit from purified, active enzyme. Amino acid sequence and composition of the C-terminal tryptic peptide of the large subunit from purified hydrogenase revealed that it is 15 amino acids shorter than that predicted by the nucleotide sequence. Processing of the nascent large subunit occurs by C-terminal cleavage between His536 and Val537, residues which are highly conserved among [NiFe] hydrogenases. Mutagenesis of the analogous residues, His582 and Val583, in the E. coli hydrogenase-1 (HYD1) large subunit resulted in significant decrease in processing and HYD1 activity. PMID- 8405420 TI - Adenosine stimulates 5'-nucleotidase activity in rat mesangial cells via A2 receptors. AB - Because A2 adenosine receptor activation stimulates adenylate cyclase and cyclic AMP induces 5'-nucleotidase expression in rat mesangial cells, we examined the effect of adenosine and its analogs on 5'-nucleotidase activity in these cells. A2 adenosine receptors were characterized using [3H]5'-N ethylcarboxamidoadenosine (NECA) as a tracer. There was a single group of receptor sites with a KD value of 0.53 microM and a number of sites of 1,317 fmol/mg. [3H]NECA binding was inhibited preferentially by A2 adenosine analogs and antagonists. Similarly, the order of potency for cAMP stimulation was in favour of A2 adenosine analogs. Rat mesangial cells expressed surface 5' nucleotidase activity. Exposure of cells for 48 h to adenosine analogs showed that at low concentrations A2 analogs stimulated 5'-nucleotidase activity. These results indicate that adenosine upregulates activity of 5'-nucleotidase, the enzyme responsible for its local formation, via A2 receptor stimulation and increase in cAMP production. PMID- 8405421 TI - Molecular evolution of P450 superfamily and P450-containing monooxygenase systems. AB - This paper reviews the classification of the P450 superfamily which is mainly based on sequence homology. The widely accepted classification by Nebert et al. [(1991) DNA Cell Biol. 10, 1-14] as well as the results of a 'two-step' multiple sequence alignment technique show that the molecular evolution of P450s, in contrast to that of many protein families, does not follow phylogeny. The data suggest that during the evolution of P450s, gene duplications and gene fusions, horizontal gene transfer and intron loss events have occurred. 'Weak' and 'strong' hierarchies in the clustering of P450 sequences were revealed. A novel evolutionary tree of the P450 superfamily has been constructed using a multiple alignment of consensus sequences. The simple classification of known P450 containing monooxygenase systems into three-, two- and one-component systems is further discussed. Particularly, the multidomain enzyme, nitric oxide synthase (NOS), should be classified as an example of a eukaryotic one-component P450 system since its N-terminal (haem) domain exhibits similarity with microsomal P450s. PMID- 8405422 TI - Endoproteolysis of non-CAAX-containing isoprenylated peptides. AB - A microsomal endoprotease specifically cleaves isoprenylated peptides of the CAAX motif, such as N-acetyl-S-all-trans-farnesyl-L-cysteine (AFC-VIM), at the isoprenylated cysteine residue. It is shown here that endoproteolysis will also occur with peptides which are not of the CAAX type. Peptide substrates modeled after the Delta virus large antigen carboxyl-terminus (CRPQ) are endoproteolytically hydrolyzed by liver microsomes. AFC-RPQ is hydrolyzed with a KM = 12.4 microM and a Vmax = 0.27 nmol/min/mg, and AGGC-RPQ is hydrolyzed with a KM = 7.9 microM and a Vmax = 0.042 nmol/min/mg. Moreover, a series of potent inhibitors of the endoproteolysis of AFC-AAX-containing peptides are ineffective at inhibiting the hydrolysis of AFC-RPQ and AGGC-RPQ, suggesting the existence of isoforms of the endoprotease. PMID- 8405423 TI - Distinct expression of two Drosophila homologs of fibroblast growth factor receptors in imaginal discs. AB - The expression of two Drosophila homologs of FGF receptors (DRF1 and DRF2) in imaginal discs was studied. DFR1 mRNA was observed in several imaginal discs, whereas DFR2 mRNA was not detected. DFR1 expression in the wing and leg discs took place in probable myoblasts in a pattern similar to that of twist, a mesodermal gene. The mRNA was also detected in the morphogenetic furrow and its posterior region of the eye disc and around the proliferation center of the brain. These results suggest that DFR1 is involved in the development of mesodermal and neuronal cells constituting the adult body. PMID- 8405424 TI - 27-Hydroxycholesterol modulation of low density lipoprotein metabolism in cultured human hepatic and extrahepatic cells. AB - 27-Hydroxycholesterol**, 25-hydroxycholesterol and cholesterol suppressed LDL uptake and degradation in human extrahepatic and hepatic cell lines in a concentration-dependent manner. Cholesterol was the least potent, and the inhibitory effect of oxysterols was more pronounced in skin fibroblasts and in endothelial cell line EAhy 926 than in hepatoma HepG2 cells. Shorter incubations were required for oxysterols to achieve 50% inhibition of LDL uptake and degradation in all cultured cells. The inhibition of LDL catabolism in extrahepatic cells by 27-hydroxycholesterol occurred at concentrations close to those observed in human plasma (0.2-0.6 microM). The results support a possible role of 27-hydroxycholesterol, a physiological oxysterol, in the regulation of cellular cholesterol homeostasis in non-hepatic tissues. PMID- 8405425 TI - A change in soybean agglutinin binding patterns of bovine milk fat globule membrane glycoproteins during early lactation. AB - Milk fat globule membrane (MFGM) glycoproteins were prepared from bovine milk at different stages of early lactation. Western blot analyses using several lectins revealed that reactivity of MFGM glycoproteins, especially 47K and 80K bands, to soybean agglutinin (SBA) remarkably increased during the lactation, while no change was observed for Ricinus communis agglutinin-I (RCA-I) binding. Sialidase treatment of MFGM glycoproteins revealed that the number of SBA-positive bands and the amount of SBA-positive oligosaccharides in these bands are increased during the lactation. Since SBA binds N-acetylgalactosamine terminated oligosaccharides, the results indicated that N-acetylgalactosaminylation of bovine MFGM glycoproteins is stimulated during the lactation. PMID- 8405426 TI - Autoimmune Addison's disease. Analysis of autoantibody binding sites on human steroid 21-hydroxylase. AB - Human steroid 21-hydroxylase (21-OH) expressed in an in vitro translation system was found to react specifically with adrenal autoantibodies from patients with Addison's disease. The epitopes on 21-OH which reacted with autoantibodies were studied by incorporating a series of terminal and internal deletions into the 21 OH gene and analysing the expressed proteins by Western blotting. N-Terminal deletions up to amino acid 280 had no effect on autoantibody binding whereas a series of C-terminal deletions and truncations (amino acids 281-494) showed marked effects. Our results indicate that a central segment (281-379) and a C terminal segment (380-494) of 21-OH interact to form at least one major autoantibody binding site. PMID- 8405427 TI - Assay conditions for the mitochondrial NADH:coenzyme Q oxidoreductase. AB - The assay of Complex I activity requires the use of artificial acceptors, such as short-chain coenzyme Q homologs and analogs, because the physiological quinones, such as CoQ10, are too insoluble in water to be added as substrates to the assay media. The medical interest raised in the last years on the pathological changes of Complex I activity has focussed on the requirement of easy reliable assays for its analysis. We have undertaken a systematic examination of the assay conditions of Complex I in mitochondrial membranes, using a series of quinones as electron acceptors, particularly the coenzyme Q homologs CoQ0, CoQ1 and CoQ2, and the analogs duroquinone and decylubiquinone. Our findings have pointed out that the most suitable electron acceptor for the NADH:CoQ reductase assay is the homolog CoQ1. The analog DB, commercially available, although yielding a high activity, nevertheless causes some problems for the standardization of the assay conditions. PMID- 8405428 TI - Characterization of the 97 and 103 kDa forms of starch branching enzyme from potato tubers. AB - N-Terminal analysis, peptide mapping and partial peptide sequencing of the 97 and 103 kDa forms of starch branching enzyme from potato tubers showed that the two forms are highly related. A comparison with sequence data in the literature showed that these forms belong to the starch branching enzyme isoform I family. An internal cDNA fragment was obtained using PCR technology on potato tuber RNA with two oligonucleotide primers constructed from the peptide sequence data. Southern blot analysis using the PCR fragment as probe showed that there is only one gene locus encoding this isoform of the enzyme in Solanum tuberosum as well as in Solanum commersonii. PMID- 8405429 TI - Residues 1 to 80 of the N-terminal domain of the beta subunit confer neuronal bungarotoxin sensitivity and agonist selectivity on neuronal nicotinic receptors. AB - Standard two electrode voltage clamp techniques were used to investigate the response of neuronal nicotinic acetylcholine receptors, expressed in Xenopus oocytes, to various agonists and neuronal bungarotoxin (NBT). The beta subunit is an important determinant of the receptor's pharmacological profile. Co-expression of alpha 4 and beta 2 subunits produced a receptor that was relatively insensitive to cytisine and nicotine and inhibited by NBT, whilst the alpha 4 beta 4 combination produced a receptor that was highly sensitive to cytisine and nicotine but resistant to toxin. The first 80 amino acids of the N-terminal domain of the beta subunit are implicated in these characteristics, since the combination of alpha 4 with a hybrid beta subunit comprising amino acids 1-->80 of beta 2 and 81-->416 of beta 4 became relatively insensitive to nicotine and cytisine and resistant to inhibition by neuronal bungarotoxin. PMID- 8405430 TI - Differential expression of AMPA glutamate receptor mRNAs in the rat adrenal gland. AB - Localisation of mRNA encoding the four AMPA glutamate receptor subunits in the rat adrenal gland was investigated using in situ hybridisation. GluR1 is found in the zona glomerulosa of the cortex, GluR3 in the remaining parts of the cortex, GluR2 in adrenal medullary cells and GluR4 at a very low level in the zona glomerulosa. All four receptor mRNAs are found in medullary ganglion cells. The flip form of GluR2 and GluR3 dominate and the GluR2 mRNA is present in the arginine encoding form. Different cell populations of the adrenal gland may express homomeric forms of different receptor subtypes. PMID- 8405431 TI - Topological characterization and modeling of the 3D structure of lipase from Pseudomonas aeruginosa. AB - Lipase from Pseudomonas aeruginosa is a M(r) 29 kDa protein with a single functional disulfide bond as shown by a shift in electrophoretic mobility after treatment with dithiothreitol and iodoacetamide. Limited proteolysis of lipase with Staphylococcus aureus protease V8 resulted in cleavage after amino acid residues Asp38 and Glu46. Comparison of the lipase amino acid sequence with those of other hydrolases with known 3D structures indicated that the folding pattern might be compatible with the alpha/beta hydrolase fold, thereby allowing us to construct a 3D model which fitted the biochemical properties. The model predicts a catalytic triad consisting of Ser82, Asp229 and His251, and contains a disulfide bond connecting residues Cys183 and Cys235. Residues Asp38 and Glu46 are located at the surface of the enzyme, whereas the disulfide bond is rather inaccessible, which is in agreement with the finding that the protein needed to be partly unfolded before a reduction of the disulfide bond could take place. A striking prediction from the model was the lack of a lid-like alpha-helical loop structure covering the active site which confers to other well-characterized lipases a unique property known as interfacial activation. Experimental determination of lipase activity under conditions where the substrate existed either as monomeric solutions or aggregates confirmed the absence of interfacial activation. PMID- 8405432 TI - Solvent oxygen is not incorporated into N10-formyltetrahydrofolate in the reaction catalyzed by N10-formyltetrahydrofolate synthetase. AB - The mechanism of the reaction catalyzed by N10-formyltetrahydrofolate synthetase involves the formation of formyl phosphate as an intermediate which then formylates tetrahydrofolate at the N-10 position. Previous studies demonstrated that the non-enzymic formylation of tetrahydrofolate by formyl phosphate occurs exclusively at the more nucleophilic 5-nitrogen in the reduced pyrazine ring. The experiments described in this report were designed to determine whether N5 formyltetrahydrofolate might be the first product to be formed on the enzyme, followed by formyl transfer to the 10-nitrogen via the cyclic intermediate N5,10 methenyltetrahydrofolate. If this were the case, oxygen from solvent H2O would be incorporated into the formyl group of the N10-derivative. By conducting the reaction in a 1:1 mixture of [16O]H2O and [18O]H2O and using 13C NMR spectroscopy we show that no 18O is incorporated into the product and conclude that the reaction proceeds via a direct formylation of the N-10 position by formyl phosphate. PMID- 8405433 TI - Characterization of the P4 promoter region of the human insulin-like growth factor II gene. AB - The human insulin-like growth factor II (IGF-II) gene contains four promoters (P1, P2, P3 and P4). In order to determine the mechanism by which the P4 promoter is controlled, the human IGF-II P4 promoter was analyzed in cell lines. DNA sequence analysis of the human IGF-II P4 promoter gene showed that the P4 promoter region contains a TATA-like sequence and several G+C rich regions which are essential for transcription. Analysis of the transcription initiation site by S1 nuclease mapping revealed two transcription start sites; both are located immediately behind TATA-like sequence. To determine the location of sites that may be important for the function of the human IGF-II P4 promoter, we constructed chimeric genes of the human IGF-II P4 promoter fused to the coding region for chloramphenicol acetyltransferase (CAT). These constructs were transfected into HepG2, PLC/PRF/5, G401 and A549 cells, and were examined for CAT activity. All transfected cells showed a similar profile of CAT activity. Sequences responsible for putative enhancer and silencer regions were identified and the 5' flanking sequences of the human IGF-II P4 promoter contain negative regulatory regions ( 213 to -174). The 53-base pair fragment located between 111 and 59 base pairs upstream of the start site contains positive regulatory activity. Gel mobility shift assay showed that Sp1 and another proteins might be involved in positive regulation of the human IGF-II P4 promoter. PMID- 8405434 TI - Carcinogenic antioxidants. Diethylstilboestrol, hexoestrol and 17 alpha ethynyloestradiol. AB - The synthetic oestrogens diethylstilboestrol, hexoestrol and 17 alpha ethynyloestradiol are known to be carcinogenic, yet they all exert antioxidant properties in vitro in that they are good inhibitors of iron ion-dependent lipid peroxidation. In rat liver microsomes incubated with Fe(III)-ascorbate or Fe(III) ADP/NADPH and also in ox-brain phospholipid liposomes incubated with Fe(III) ascorbate; the overall order of effectiveness of the compounds tested as inhibitors of lipid peroxidation was diethylstilboestrol > hexoestrol > 17 alpha ethynyloestradiol > 4-hydroxytamoxifen > 17 beta-oestradiol > tamoxifen. Compounds acting as antioxidants towards lipids may also exert pro-oxidant effects towards other molecules such as DNA and thus must never be assumed to be safe for human use. PMID- 8405435 TI - Induction of phosphatidic acid by fibroblast growth factor in cultured baby hamster kidney fibroblasts. AB - Basic fibroblast growth factor (bFGF/FGF-2) is a strong mitogenic inducer of cultured baby hamster kidney (BHK) cells. When cultured BHK cells were stimulated with FGF-2, phosphatidic acid (PA) was induced within 2 min, peaked at 5 min and gradually decreased. Phospholipase D (PLD) was also mitogenic for cultured BHK cells and this effect was mediated via PA. The possibility that PA induction by FGF-2 is an essential signaling step for BHK cell proliferation is discussed. PMID- 8405436 TI - Protein kinase C alpha, delta, epsilon and zeta in C6 glioma cells. TPA induces translocation and down-regulation of conventional and new PKC isoforms but not atypical PKC zeta. AB - Both the cytosol and membrane in C6 glioma cells express abundance of PKC alpha, delta, zeta and trace amount of PKC epsilon by Western blot analysis with isozyme specific antibodies. These characteristics make this cell line a good model to study the properties of different classes of PKC isoforms in one cell type. Exposure of the cells to 100 nM TPA for 10 min resulted in the translocation of conventional PKC alpha (cPKC alpha) and new PKC delta (nPKC delta) and -epsilon from the cytosolic to the membrane fraction, while left atypical PKC zeta (aPKC zeta) unaffected. The extent of translocation of cPKC alpha induced by TPA was more prominent than that of nPKC delta and nPKC epsilon. alpha-TPA, the inactive phorbol ester, did not induce translocation of these isozymes. After treatment of the cells with 1 microM TPA for 17 h, cPKC alpha, nPKC delta and nPKC epsilon were almost completely down-regulated, whereas aPKC zeta was still unaffected. The natural activators of this cell line, endothelin-1 and ATP also translocated cPKC alpha and nPKC delta. However, the extent of translocation induced by these two agonists was much less than that of TPA. PMID- 8405437 TI - Differential shedding of the two subunits of the interleukin-6 receptor. AB - cDNAs coding for the two receptor subunits of the interleukin-6 receptor have been stably expressed in Madine Darby canine kidney (MDCK) cells. The fate of the IL-6 binding protein (IL-6R) and of the signal transducing protein gp130 was studied independently. Both proteins were proteolytically cleaved from cells metabolically labeled with [35S]methionine/cysteine leading to the release of soluble receptor proteins of 55 kDa and 100 kDa, respectively. In contrast to the shedding of the IL-6R gp130 was inefficiently released from the cells and the process was not significantly stimulated by the phorbolester PMA. In addition we show that the soluble forms of the IL-6R and gp130 released by transfected cells can form a ternary complex with interleukin-6 indicating that such complexes also may occur in vivo. PMID- 8405438 TI - Improved gene transfer by direct plasmid injection associated with regeneration in mouse skeletal muscle. AB - Gene transfer into skeletal muscle via simple plasmid injection in vivo has many potential uses but these are severely constrained by the low efficiency of this technique. Muscle regeneration, induced by the myotoxic local anaesthetic bupivacaine, significantly increased gene expression following plasmid injection 3-7 days after bupivacaine treatment. Much of this effect can be attributed to uptake and expression of the plasmid by a greater number of muscle fibres, up to 9% of the mouse tibialis anterior muscle. A similar significant increase in expression was observed in the naturally regenerating muscle of the dystrophic mdx mouse when compared to the control C57Bl/10 strain. PMID- 8405439 TI - Characterisation of biotinylated liposomes for in vivo targeting applications. AB - Liposomes containing monosialoganglioside (GM1) or polyethylene glycol (PEG) lipid derivatives have prolonged circulation in the blood. This favours liposome extravasation to tumour sites. In this report it is shown that inclusion of GM1, PEG550-DPPE or PEG2000-DPPE in liposomes containing biotin-DPPE significantly diminished the ability of vesicles to bind to streptavidin in vitro. Steric inhibition due to the bulky head group of these lipids was least for biotin-DPPE liposomes containing GM1. Biodistribution studies in C26 tumour-bearing mice showed that GM1-liposomes containing small amounts of biotin-DPPE have long circulation life-times in the blood. Using fluorescent microscopic techniques, liposomes containing both GM1 and biotin-DPPE were detected within extra-vascular spaces in tumours. In addition it was shown that biotin-DPPE in GM1-liposomes bound streptavidin in situ. These results suggest that GM1-liposomes containing biotin-DPPE have potential use as diagnostic or therapeutic reagents in pre targeting applications dependent on the high-affinity interaction of biotin with streptavidin. PMID- 8405440 TI - Triple helix formation at A8XA8.T8YT8. AB - We have examined the formation of DNA triple helices between the oligonucleotides T8XT8 (X = A,C,G,T) and DNA fragments containing the target sequences A8XA8.T8YT8 (X = T,C,G; Y = A,G,C), by DNase I footprinting. We find that A8GA8.T8CT8 yields a footprint with T8CT8 and shows a weaker interaction with T17 and T8GT8. A8CA8.T8GT8 yields a footprint with T17, and shows weaker interaction with T8CT8. A8TA8.T8AT8 yields a footprint with T8GT8 and shows weaker interaction with T17. Each of the successful complexes is characterised by enhanced DNase I cleavage at the 3' end of the purine strand of the target, as well as protection at the 5' end. We have been unable to from triplexes with third strands of the type A8XA8. PMID- 8405441 TI - Anisodamine causes acyl chain interdigitation in phosphatidylglycerol. AB - The effect of anisodamine on the structure of the gel phase and the properties of the acyl chain disordering transition of dipalmitoylphosphatidylglycerol (DPPG) has been studied through high-sensitivity differential scanning calorimetry (DSC) and fluorescence polarization measurements of 16-(9-anthroyloxyl)-palmitic acid (16AP) and 3-(9-anthroyloxyl)-stearic acid (3AS), labeling, respectively, the ends and the third carbon of the acyl chains. The non-interdigitated DPPG multilamellar vesicles formed in HEPES buffer show clear fluidity gradient in their acyl chains, whereas the fluidity gradients are completely abolished in the presence of anisodamine. The DSC results showed that the phase transition temperature (Tm) of DPPG is decreased and the enthalpy (delta H) is increased by anisodamine, while the pre-transition vanishes. At 3 mM anisodamine, the delta H of DPPG reaches 9.6 kcal/mol. It can be concluded that DPPG forms an interdigitated gel phase in the presence of anisodamine. A molecular scheme for the interaction of anisodamine with DPPG is proposed. PMID- 8405442 TI - Chemical structure of lipid A from Porphyromonas (Bacteroides) gingivalis lipopolysaccharide. AB - The novel chemical structure of Porphyromonas (Bacteroides) gingivalis strain 381 lipid A was determined to be a glucosamine beta-(1-6) disaccharide 1 monophosphate acylated by 3-hydroxy-15-methylhexadecanoic acid and 3 hexadecanoyloxy-15-methylhexadecanoic acid at the 2- and 2'-positions, respectively. The absence of ester-linked phosphate at the 4'-position and fatty acids at the 3- and 3'-positions, and the presence of fatty acids possessing 16 17 carbon atoms are unique features, differentiating the lipid A from enterobacterial and other lipid As. These structural features may be related to its low endotoxic activity. Furthermore, P. gingivalis lipid A as well as its LPS stimulated the splenocytes from C3H/HeN and C3H/HeJ mice. PMID- 8405443 TI - Peroxisome proliferators and T3 operate by way of distinct receptors. AB - Peroxisome proliferators and thyroid hormones have a number of common metabolic effects. The possibility that the signal transduction pathways of both groups of effectors converge at the receptor level was investigated. It was shown that T3, specifically bound to the rat thyroid beta-receptor, was not displaced to a significant extent by ciprofibrate or bezafibrate. No specific binding of T3 to the mouse peroxisome proliferator activated receptor could be demonstrated. In transactivation experiments peroxisome proliferators were unable to activate the thyroid receptor and T3 did not activate a chimeric receptor containing the ligand binding domain of the peroxisome proliferator activated receptor. It is concluded that peroxisome proliferators and thyroid hormone do not cross-react at the level of their nuclear receptors. PMID- 8405444 TI - Increased production of endothelin-1 in the hypertrophied rat heart due to pressure overload. AB - Endothelin-1 (ET-1) has been demonstrated to induce hypertrophy in cultured cardiac myocytes. We investigated the production of ET-1 in the heart of aorta banded rats in vivo. Seven days after the banding of the abdominal aorta, rats developed a significant left ventricular hypertrophy. The tissue content of mature ET-1 and the level of expression of prepro ET-1 mRNA were higher in the left ventricle of aorta-banded rats than in those of sham-operated rats. The expression of prepro ET-1 mRNA in the right ventricle was not different between the two groups. These findings indicate that the production of ET-1 increased in the hypertrophied left ventricle, thereby suggesting the possible involvement of endogenous ET-1 in the development of cardiac hypertrophy due to pressure overload. PMID- 8405445 TI - Kinetic analysis on the substrate specificity of 3-isopropylmalate dehydrogenase. AB - Substrate specificity of 3-isopropylmalate dehydrogenase is analyzed using a series of synthetic (2R,3S)-3-alkylmalates. Each analog with hydrogen, methyl, ethyl, isopropyl, isobutyl, tert-butyl, and isoamyl group on C-3 functions as a substrate, implying a broad substrate specificity of the enzyme toward alkylmalates. The incremental binding energy of the isopropyl group of 3 isopropylmalate to the enzyme is estimated to be 3.55 kcal/mol, the rather small value supporting the broad specificity. Although the enzyme shows a broad specificity toward the alkylmalates, it does not show activity with isocitrate which has a negatively charged carboxymethyl group instead of the alkyl groups. PMID- 8405446 TI - Tyr-139 in Thermus thermophilus 3-isopropylmalate dehydrogenase is involved in catalytic function. AB - The role of Tyr-139, which is thought to be located at the active site of Thermus thermophilus HB8 3-isopropylmalate dehydrogenase, has been investigated by site specific replacement with phenylalanine. The replacement scarcely affected the Michaelis constant (Km) for 3-isopropylmalate, but caused a 13-fold decrease of that for NAD. The catalytic constant (kcat) showed a 14-fold decrease. Accordingly, the catalytic efficiency (kcat/Km) decreased for 3-isopropylmalate but not for NAD. The results suggest that Tyr-139 is involved in the catalytic function through interaction with 3-isopropylmalate. PMID- 8405447 TI - The DsbA-DsbB system affects the formation of disulfide bonds in periplasmic but not in intramembraneous protein domains. AB - The DsbA and DsbB proteins of Escherichia coli are involved in facilitating the formation of disulfide bonds in periplasmic proteins. Here, we show that the rate of formation of a disulfide bond in the periplasmic domain of the inner membrane protein leader peptidase is reduced in dsbA and dsbB strains, whereas the rate of formation of a disulfide bond engineered into the membrane embedded domain of the same protein is completely unaffected by these mutations. We conclude that the Dsb proteins do not facilitate the formation of intramembraneous disulfides. PMID- 8405448 TI - cDNA cloning of rat proteasome subunit RC7-I, a homologue of yeast PRE1 essential for chymotrypsin-like activity. AB - The nucleotide sequence of a cDNA that encodes a new subunit, named RC7-I, of the 20 S proteasome of rat hepatoma cells has been determined. The polypeptide predicted from the open reading frame consists of 201 amino acid residues with a calculated molecular weight of 22,912 and isoelectric point of 7.25. Approximately 80% of the partial amino acid sequences of several fragments of RC7 I, determined by protein chemical analyses, were found to be in excellent accordance with those deduced from the cDNA sequence. Computer analysis showed that RC7-I belongs to the beta-type subgroup of proteasomes with similarity to the beta-subunit of the archaebacterial proteasome, differing clearing from alpha type subunits of the proteasome gene family. The overall structure of RC7-I was found to be homologous to that of yeast PRE1, which is necessary for chymotryptic activity. PMID- 8405449 TI - Identification of a new b-type cytochrome from the whitefish Coregonidae eggs. AB - A new b-type cytochrome, termed cytochrome b560 according to the wavelength maximum of its alpha-band in the reduced minus oxidized difference spectrum, was isolated from eggs of whitefish and partially purified by fractionation of the water-soluble moiety of yolk. The absolute absorption spectrum of reduced cytochrome b560 has maxima at 426, 529 and 560 nm and that of the oxidized form at 416 nm. The reduced minus oxidized difference spectrum has maxima at 428, 529 and 560 nm. The midpoint potential of this cytochrome is +193 mV. Based on the MCD spectra of reduced cytochrome and the optical absorption spectra in the visible region of the oxidized cytochrome, it is suggested that the heme iron in cytochrome b560 has two histidine imidazoles as the 5th and 6th axial ligands. PMID- 8405450 TI - Calorimetric characterization of the stable complex of myosin subfragment 1 with ADP and beryllium fluoride. AB - The thermal unfolding of the myosin subfragment 1 (S1) in its stable complex with ADP and beryllium fluoride (S1.ADP.BeF3-) was studied by differential scanning calorimetry. It has been shown that the structure of the S1 molecule in the S1.ADP.BeF3- complex is similar to that of S1 in its complex with ADP and orthovanadate (S1.ADP.Vi) but differs radically from that of nucleotide-free S1 and S1 in the S1.ADP complex. It is concluded that the S1.ADP.BeF3- complex can be considered, like the S1.ADP.Vi complex, a stable structural analogue of the myosin head.ADP.Pi transition state of the myosin-catalyzed ATP hydrolysis. PMID- 8405451 TI - pH dependence of the formation of an M-type intermediate in the photocycle of 13 cis-bacteriorhodopsin. AB - An M-type intermediate is formed in the 13-cis-bR photocycle in purple membranes at high pH. This is presumably due to deprotonation of the same group whose deprotonation causes a large increase in rate of M formation in the trans-bR photocycle (the 'alkaline transition'). For Triton X-100-solubilized bR, the alkaline transition is shifted to a lower pH value by more than 2 pH units. The alkaline transition in Triton-solubilized preparations changes the efficiency of the M intermediate formation in the 13-cis-sbR photocycle. The M intermediate formation in 13-cis-sbR, as in the case of trans-sbR, is completely inhibited when the blue 'acidic' bR is formed at low pH. The protonation state of the group affecting formation of the M intermediate in 13-cis-bR at high pH and the group which is responsible for the transition to the blue acidic form influence in a similar way the equilibrium between bR isomers in the dark-adapted form as well as the rate of dark adaptation. PMID- 8405452 TI - Yeast coatomer contains a subunit homologous to mammalian beta'-COP. AB - The homologue of the mammalian coatomer complex was isolated from yeast cytosol and separated on a modified urea-containing SDS-polyacrylamide gel system. An additional band in the 100 kDa molecular weight range appeared when compared to the protein pattern obtained in conventional Laemmli gels, exactly as observed for mammalian coatomer. Cross-reactivity with an anti-peptide antibody raised against the C-terminus of beta'-COP from bovine, and N-terminal sequence analysis, revealed that this protein from yeast is related to beta'-COP from mammals. PMID- 8405453 TI - Cloning, sequence and expression of the gene encoding the malolactic enzyme from Lactococcus lactis. AB - Many lactic acid bacteria can carry out malolactic fermentation. This secondary fermentation is mediated by the NAD- and Mn(2+)-dependent malolactic enzyme, which catalyses the decarboxylation of L-malate to L-lactate. The gene we call mleS, coding for malolactic enzyme, was isolated from Lactococcus lactis. The mleS gene consists of one open reading frame capable of coding for a protein with a calculated molecular mass of 59 kDa. The amino acid sequence of the predicted MleS gene product is homologous to the sequences of different malic enzymes. Bacterial and yeast cells expressing the malolactic gene convert L-malate to L lactate. PMID- 8405454 TI - Assignment of the backbone 1H and 15N NMR resonances and secondary structure characterization of barstar. AB - Barstar, a polypeptide inhibitor of ribonucleases, has been studied by 2D and 3D NMR techniques using uniformly 15N-labeled protein. Backbone (15NH-C alpha H-C beta H) resonances were assigned for all but 5 of the 89 residues. Dihedral angle and deuterium exchange studies were used in conjunction with medium range inter proton NOEs to characterize the secondary structure of barstar. The protein is composed of four alpha-helices and three short stretches of extended strand. By further analysis of the NOE data three of the helices were found to be parallel to each other with the single disulphide bond linking the second and fourth helices at their C-terminal ends. PMID- 8405455 TI - Differential modulation of pancreatic beta-cell bursting by intracellular pH in the presence and absence of a K-ATP channel blocker. AB - The study of the influence of intracellular pH (pHi) changes on the mechanism underlying pancreatic beta-cell bursting has been hampered by concomitant effects on the activity of background ATP-dependent K+ (K-ATP) channels. beta-cells were made to burst in the absence of active K-ATP channels by raising external Ca2+ in the presence of 11 mM glucose and tolbutamide. An alkalinizing pHi shift (exposure to 20 mM NH4Cl) increased the burst active phase duration. Conversely, an acidifying shift (NH4Cl withdrawal) suppressed the electrical activity. This is the mirror image of the effects recorded in the absence of tolbutamide. Glibenclamide and quinine suppressed the alkalinization-evoked hyperpolarization. This study emphasizes the differential sensitivity of different beta-cell ion channels to pHi and the prevalent role of K-ATP channels as electrical transducers of cytoplasmic pH changes under regular physiological conditions. PMID- 8405456 TI - Prevention of the induced synthesis and secretion of group II phospholipase A2 by brefeldin A. AB - Brefeldin A (BFA) has previously been shown to block protein secretion and to cause dismantling of the Golgi cisternae in many cultured cell lines. BFA was found to prevent the induced synthesis and secretion of 14 kDa group II phospholipase A2 (PLA2) in rat mesangial cells. Furthermore, BFA inhibited total protein synthesis although PLA2 appeared to be more sensitive to the effect of this compound than total protein synthesis assessed by amino acid incorporation. BFA was unable to block protein synthesis or PLA2 activity in the cell completely but secretion of enzymatic activity and PLA2 protein into the cell culture media was totally inhibited. PMID- 8405457 TI - Redox control of transcription: sensors, response regulators, activators and repressors. AB - In a growing number of cases, transcription of specific genes is known to be governed by oxidation or reduction of electron carriers with which the gene products interact. The biological function of such control is to activate synthesis of appropriate redox proteins, and to repress synthesis of inappropriate ones, in response to altered availability of specific electron sources and sinks. In prokaryotic systems this control appears to operate by two general classes of mechanism: by two-component regulation involving protein phosphorylation on histidine and aspartate; and by direct oxidation-reduction of gene repressors or activators. For the first class, termed 'two-component redox regulation', the term 'redox sensor' is proposed for any electron carrier that becomes phosphorylated upon oxidation or reduction and thereby controls phosphorylation of specific response regulators, while the term 'redox response regulator' is proposed for the corresponding sequence-specific DNA-binding protein that controls transcription as a result of its phosphorylation by one or more redox sensors. For the second class of redox regulatory mechanism, the terms 'redox activator protein' and 'redox repressor protein' are proposed for single proteins containing both electron transfer and sequence-specific DNA-binding domains. The structure, function and biological distribution of these components are discussed. PMID- 8405458 TI - 2-Oxo-histidine as a novel biological marker for oxidatively modified proteins. AB - We report a promising finding that oxidative modification of proteins by free radicals could be monitored by the formation of oxidized histidine that is detectable by reverse-phase HPLC with electrochemical detection (HPLC-ECD). When the N-protected histidine derivative (N-benzoylhistidine) was exposed to a free radical-generating system (copper/ascorbate), a number of products were detected by HPLC-ECD and the main product among them was found to be identical to N benzoyl-2-oxo-histidine. The acid hydrolysis of N-benzoyl-2-oxo-histidine provided a single product (2-oxo-histidine) that was detected sensitively by HPLC ECD. Thus 2-oxo-histidine was indeed generated as the main product in the oxidatively modified proteins by free radicals. Taken together, 2-oxo-histidine may be a useful biological marker for assessing protein modifications under oxidative stress. PMID- 8405459 TI - Prenylated protein methyltransferase of rat cerebellum is developmentally co expressed with its substrates. AB - High levels of prenylated protein methyltransferase are expressed in the developing rat cerebellum and are responsible for methylation of endogenous G proteins and 50-52 kDa synaptosomal proteins. Enzyme activity in cerebellar synaptosomes of 3 week postnatal rats is 2-fold higher than that found in adult rat cerebellum. A 10-fold rise in activity occurs at the end of the second and during the third postnatal weeks, followed by a subsequent decline. Expression of the enzymes' substrates follows the same pattern. The high methyltransferase activity in 3-week-old cerebellum coincides with the period of granule cell migration and synaptogenesis, suggesting a regulatory role for the enzyme and its substrates in cerebellar ontogenesis. PMID- 8405460 TI - Fragmentation of DNA in P388D1 macrophages exposed to oxidised low-density lipoprotein. AB - Exposing a macrophage-like murine cell line to copper-oxidised low-density lipoprotein led to DNA fragmentation which was inhibited by the putative Ca2+/Mg2+ endonuclease inhibitor, zinc sulphate. DNA fragmentation preceded loss of membrane impermeability. These results suggest that apoptosis may be a mechanism of macrophage foam cell death in atherosclerotic lesions in the arterial wall. PMID- 8405461 TI - Mass spectrometric evidence for a disulfide bond in aequorin regeneration. AB - Tryptic digests of purified recombinant apoaequorin were analyzed, before and after reduction with DTT, by fast atom bombardment mass spectrometry. The results showed that apoaequorin contains a disulfide bond between Cys145 and Cys152 and that the reduction of this bond is involved in the regeneration of aequorin. PMID- 8405462 TI - Characterisation of ATP binding inhibition to the sarcoplasmic reticulum Ca(2+) ATPase by thapsigargin. AB - The inhibition of Ca(2+)-ATPase of sarcoplasmic reticulum by thapsigargin has been reported to be associated with a suppression of calcium binding to the high affinity transport sites. We report here that thapsigargin also acts as an inhibitor of ATP binding by reducing its apparent affinity by about two orders of magnitude. This inhibition is non-competitive indicating that thapsigargin does not bind to the ATP binding site. This is confirmed by the fact that thapsigargin binding to the Ca(2+)-ATPase does not affect the binding of 2',3'-O-(2,4,6 trinitrocyclohexadienylidene)-ATP (TNP-ATP). PMID- 8405463 TI - Aspirin induces non-enzymatic formation of platelet-activating factor from lyso platelet-activating factor. AB - Substantial amounts of platelet-activating factor (PAF) were formed when lysoPAF was mixed with aspirin (e.g. 0.04% of added lysoPAF (200 nmol) was converted to PAF when mixed with aspirin (2 mumol) for 1 h). Non-enzymatic formation of PAF from aspirin and lysoPAF also occurs in the aqueous solution or in organic solvents in time-dependent and dose-dependent manners. Possible meanings of the non-enzymatic formation of PAF are discussed. PMID- 8405464 TI - Enhancement of Ca2+ release channel activity by phosphorylation of the skeletal muscle ryanodine receptor. AB - The Ca2+ release channel of rabbit skeletal muscle sarcoplasmic reticulum (SR) can be phosphorylated by membrane associated protein kinase(s) utilizing endogenously synthesized or exogenously added ATP. The channel protein has been enriched in non-phosphorylated and phosphorylated form from heavy SR following solubilization with CHAPS (3-[(3-cholamidopropyl)dimethylammonio-1-propane sulfonate) and ultracentrifugation on a linear sucrose/CHAPS gradient. Reconstitution of the isolated channels into planar bilayers shows that phosphorylation enhances the open probability by increasing the sensitivity towards micromolar Ca2+ and ATP. The phosphorylation induced enhancement of the channel activity can be reversed by purified protein phosphatase 2A. PMID- 8405465 TI - Purification and antipathogenic activity of lipid transfer proteins (LTPs) from the leaves of Arabidopsis and spinach. AB - Two homogeneous proteins active in vitro against the bacterial pathogen Clavibacter michiganensis subsp. sepedonicus were obtained from a crude cell-wall preparation from the leaves of Columbia wild-type Arabidopsis. The N-terminal amino acid sequences of these proteins allowed their identification as lipid transfer proteins (LTP-a1, LTP-a2); the LTP1-a1 sequence was identical to that deduced from a previously described cDNA (EMBL M80566) and LTP-a2 was quite divergent (44% identical positions). These proteins were not detected in the cytoplasmic fraction by Western-blot analysis. Proteins LTP-s1 and LTP-s2 were similarly obtained from spinach leaves; LTP-s1 was 91% identical to a previously purified spinach LTP (Swiss Prot P10976), and LTP-s2 was moderately divergent (71% identical positions). About 1/3 of the total LTPs were detected in the cytoplasmic fraction from spinach by Western-blot analysis. Concentrations of these proteins causing 50% inhibition (EC-50) were in the 0.1-1 microM range for the bacterial pathogens C. michiganensis and Pseudomonas solanacearum and close to 10 microM for the fungal pathogen Fusarium solani. PMID- 8405466 TI - Role of isoprenylation in intracellular pH regulation of granulocytes. AB - Lovastatin was used to study the role of isoprenylated GTP-binding proteins in the regulation of intracellular pH in granulocytic HL-60 cells. The cytosolic acidification that accompanies stimulation by chemoattractants was completely eliminated by lovastatin. The subsequent activation of Na+/H+ exchange was partially inhibited by the isoprenylation antagonist. In contrast, the osmotic activation of the Na+/H+ antiport was unaffected. The data indicate that the osmotically- and receptor-induced activation of the antiport utilize divergent signalling pathways. The results also provide evidence supporting the notion that cytosolic acidification by chemoattractants results from accumulation of metabolic acid generated during the respiratory burst. PMID- 8405467 TI - In vitro activation of pro-cathepsin B by three serine proteinases: leucocyte elastase, cathepsin G, and the urokinase-type plasminogen activator. AB - In vitro activation of pro-cathepsin B purified from ascitic fluid of ovarian carcinomas by serine proteinases was studied. Both elastase and cathepsin G from human leucocytes were found to be activators, on the basis of generation of cathepsin B activity and processing of the precursor. These results represent a new cooperative pathway between cancer cells and host cells. The urokinase-type plasminogen activator activated pro-cathepsin B faster than leucocyte proteinases. A new relationship is emerging between the cysteine proteinases and the plasmin-activation system. Both pathways suggest an important role of cathepsin B in the proteolytic cascade associated with tumour invasion. PMID- 8405468 TI - The effect of sugars on the morphology of the bacterial flagellum. AB - Using dark-field microscopy, we have found that certain sugars cause the normal to-curly helical transition of bacterial flagella. Titration of flagella isolated from Salmonella typhimurium with 16 different carbohydrates showed that: (i) only certain sugars cause the transition. There is no obvious relationship between the simple physico-chemical properties of the sugar and whether the sugar causes the transition or not; (ii) the efficacies of sugars that do cause the transition differ markedly. For these sugars there is a relationship between efficacy and molecular size. These results suggest that the specific, though weak, binding of sugars to sites on flagella cause the morphological transition. PMID- 8405469 TI - Selective interaction of ferricyanide with cluster I of Clostridium pasteurianum 2[Fe4S4] ferredoxin. AB - Treatment of Clostridium pasteurianum ferredoxin (CpFd) with stoichiometric amounts of potassium ferricyanide results in the specific conversion of cluster I into a Fe3S4+ species while leaving cluster II unaltered. Ferricyanide-treated CpFd derivative has been purified and characterized through biochemical and spectroscopical techniques. The cluster conversion process is reversible and reconstitution of native CpFd has been afforded under appropriate conditions. PMID- 8405470 TI - A constitutive form of heat-shock protein 70 is located in the outer membranes of mitochondria from rat liver. AB - HSP73, the constitutive form of heat-shock protein 70, has been implicated in the translocation of preproteins across the mitochondrial membranes, being required for maintaining mitochondrial preproteins in an import competent conformation. Here we report that highly purified mitochondrial outer membranes contain a protein indistinguishable from HSP73 as a tightly associated peripheral component of the membrane. This membrane form of HSP73 was photolabelled with [alpha 32P]ATP and could be released from the outer membrane with sodium carbonate, but not after incubation of the membranes with salt or with ATP. A sensitive immunoassay with an anti-HSP73 monoclonal antibody, revealed the association of HSP73 with mitochondrial outer membrane vesicles at a level similar to that of preprotein import receptors. PMID- 8405471 TI - Molecular structure of ras-related small GTP-binding protein genes of rice plants and GTPase activities of gene products in Escherichia coli. AB - We isolated two rice cDNA clones (ric1 and ric2) encoding proteins homologous to the ras-related small GTP-binding protein. The amino acid sequences of ric1 and ric2 are conserved in four regions involved in GTP binding and hydrolysis which are characteristic in the ras and ras-related small GTP-binding protein genes. In addition, two consecutive cysteine residues near the carboxyl-terminal end required for membrane anchoring are also present in ric1 and ric2. The ric1 and ric2 proteins synthesized in Escherichia coli possessed GTPase activity (i.e. hydrolysis of GTP to GDP). PMID- 8405472 TI - The reaction of ascorbic acid with different heme iron redox states of myoglobin. Antioxidant and prooxidant aspects. AB - The interaction of ascorbate with different heme iron redox states of myoglobin (ferrylmyoglobin, FeIV = O; metmyoglobin, FeIII; and oxymyoglobin, FeIIO2) was examined by e.s.r. and absorption spectroscopy. The reaction of ascorbate with ferryl- or met-myoglobin resulted in ascorbyl radical production. The interaction of ascorbate with oxymyoglobin proceeded with formation of ascorbyl radical, hydrogen peroxide, and an overall oxidation of oxymyoglobin to metmyoglobin. The latter reaction proceeded via an oxoferryl complex intermediate-corresponding to ferrylmyoglobin and identified by treatment of the reaction mixture with Na2S. These observations are consistent with a concerted electron transfer mechanism, whereby the two electrons required for the reduction of oxygen to hydrogen peroxide are donated by ascorbic acid and the heme iron. The antioxidant and prooxidant aspects of these redox transitions are discussed in terms of their kinetic properties. PMID- 8405473 TI - Influence of illumination on the electronic interaction between 77Se and nickel in active F420-non-reducing hydrogenase from Methanococcus voltae. AB - The selenium-containing F420-non-reducing hydrogenase from Methanococcus voltae was anaerobically purified. The enzyme as isolated showed an EPR spectrum with gx,y,z = 2.21, 2.15 and 2.01. Upon illumination this spectrum disappeared and a new signal with the lowest g value at 2.05 arose. EPR studies were carried out either with the enzyme containing natural selenium or enriched in the nuclear isotope 77Se. The hyperfine splitting caused by 77Se in the 'dark' signal is shown to be highly anisotropic. In contrast the splitting is nearly isotropic after illumination. A new model for the nickel site is proposed to explain these observations. PMID- 8405474 TI - Postoperative irradiation for thyroid cancer. AB - The place of external irradiation for thyroid cancers remains controversial. Between 1974 and 1989, 94 patients with a differentiated thyroid cancer were seen in our department for additional treatment (radioiodine with or without external radiation) after a surgical resection. 56 patients were treated with radioiodine alone and 38 patients had an additional course of external radiation. Survival was identical for the two groups but the local relapse rates were 21% and 3% respectively. So, in this retrospective study, the addition of external radiation appears to improve the local control especially in the case of an incomplete surgical resection or extracapsular tumor extension. This benefit was achieved without severe radiation-induced late effects. PMID- 8405475 TI - Cancer in pregnancy. AB - OBJECTIVE: To estimate the incidence of cancer arising in pregnancy and to report its recurrence in those women with a past history of the disease. DESIGN: Retrospective study over 11 years. SETTING: John Radcliffe Maternity Hospital. PATIENTS: 25,568 Oxford District Health Authority residents who delivered at the John Radcliffe Maternity Hospital, and 6775 residents who had a termination of pregnancy, between 1 January 1981 and 31 December 1985. INTERVENTIONS: Retrospective analysis of case records to identify pregnancies complicated by cancer and follow-up through patients' general practitioners. MAIN OUTCOME MEASURES: Maternal mortality and disease recurrence. RESULTS: The study identified 32 pregnancies complicated by cancer in 28 women and four terminations of pregnancy performed for cancer as the main or secondary indication. By the end of 1991, three women had died, one woman had been treated for disease recurrence, 17 women were in good health and seven women had been lost to follow-up. There were six cases of cancer arising de novo in pregnancy, i.e. an incidence of 2.35 per 10,000 deliveries (95% confidence interval 0.47 to 4.22). Only one pregnancy was complicated by disease recurrence. CONCLUSION: The incidence of cancer arising de novo in pregnancy is lower than the most quoted figure of 9.92 per 10,000 pregnancies. PMID- 8405476 TI - Biopsy of mammographically-detected breast lesions in a district hospital. AB - Seventy-nine consecutive needle localization biopsies were performed for non palpable breast lesions discovered on screening mammography over a two-and-a-half year period in a district hospital by 15 surgeons. Malignant disease was found in 24 biopsies (30%). Eight were in situ and 16 were infiltrative carcinomas. Half the cancers met the criteria of 'minimal' carcinoma as described by Gallagher and Martin. Forty percent of women 50 years and older (20 of 50) but only 13% of women less than 50 years (4 of 29) had a carcinoma. Of thirty-two mammograms read as 'probably' benign, two (6%) were malignant pathologically. Of forty-seven mammograms considered 'probably' malignant, 22 (47%) were malignant pathologically. Abnormal calcification (38%), density (34%) or both (28%) were suspicious mammographic features. All 'minimal' invasive carcinomas were associated with calcification. Complete removal of the lesion was not achieved in three cases (3.7%) of which none showed invasive malignancy. Of the seven complications, two required treatment. Localization biopsy of non-palpable breast lesions, detected by screening mammography in a district hospital, performed by general surgeons, has produced a yield of malignancies similar to that seen in more specialized units. PMID- 8405477 TI - A review of clinicopathologic features of pheochromocytomas in Hong Kong Chinese. AB - Twenty-two Chinese patients with pheochromocytomas including 18 surgical and four autopsy cases were reported. The incidence at autopsy was 0.048%. The tumours were most common in the sixth and the fourth decades in males and females, respectively. There was no sex predilection. The incidence of bilaterality was 4.5% and the tumours were more common on the right side. 9.1% of the tumours were malignant and they were larger than their benign counterparts. Solid and diffuse pattern and mixed pattern were the most common histologic patterns. Minor histological features included hyaline globules (64%), nuclear pseudo-inclusions (55%), lipid degeneration (4.5%) and peri-adrenal brown fat (18%). All of them were strongly positive for the three pan-neuroendocrine markers (neuron-specific enolase, synaptophysin and chromogranin) confirming the usefulness of these markers in diagnosing pheochromocytoma. PMID- 8405478 TI - Magnetic resonance imaging of retroperitoneal lymph node metastases of non seminomatous germ cell tumours of the testis. AB - The value of Magnetic Resonance Imaging (MRI) in the diagnosis, staging and treatment evaluation of retroperitoneal lymph node metastases of non-seminomatous germ cell tumours of the testis (NSGCT) was prospectively studied in 41 consecutive patients before, during and after cisplatin-based chemotherapy. MRI is a non-invasive method, with results equivalent to laparotomy in determining the anatomical localization and size of retroperitoneal lymph node metastases of non-seminomatous germ cell tumours of the testis and residual mass after chemotherapy. Components responding differently can be observed with MRI within the retroperitoneal lymph node metastases on the basis of the tumour signal intensity or homogeneity in the T1- and T2-weighted images; MRI cannot warrant any conclusion about viability of the residual mass or other effects of chemotherapy. PMID- 8405479 TI - A comparative analysis of the serum levels of NCC-ST-439, CEA and CA19-9 in patients with colorectal carcinoma. AB - To investigate the usefulness of NCC-ST-439 (ST439) as a new tumor marker in colorectal cancer, we compared its serum level with the serum levels of CEA and CA19-9. The serum levels of ST439, CEA and CA19-9 from 168 patients were evaluated, of which 124 were primary cases and 44 were recurrent. ST439 was elevated in 31.5% in primary cases and 52.3% in newly-discovered carcinoma patients with confirmed recurrent cases. Following resection of the tumor, ST439 levels decreased rapidly. Similar trends in serial CEA and CA19-9 levels were found. While CEA is more sensitive for the detection of colorectal carcinomas than the other two markers, ST439 is sensitive in the detection of localized carcinomas and recurrent cases with localized lesions or liver metastasis. The results indicate that ST439 is a useful tumor marker not only for diagnosing primary carcinomas, but also for the detection of recurrent cases. PMID- 8405480 TI - Repeated hepatic resections for colorectal metastases. AB - Repeated hepatic resection (RHR) for recurrent colorectal metastases remains uncommon and controversial. We report our experience in order to assess the feasibility and the potential oncologic benefit of such an aggressive management. From 1981 to 1991, 13 patients underwent a RHR. The first hepatic resection had been an anatomic hepatectomy removing between two and six segments in 10 patients and a wedge resection in three. The RHR was performed after a mean delay of 16 +/ 10 months (5-35) from the first liver procedure. The RHR was an anatomic hepatectomy in eight patients (including a right hepatectomy in three) and a minor resection (tumorectomy or segmentectomy) in five. Three patients underwent a third liver resection for recurrence. There was no post-operative mortality. Eleven patients died from recurrence after a mean survival of 17 +/- 13 months from the second hepatic procedure (range: 6-47). One patient died from unrelated disease after 12 months and one was alive free of recurrence 22, 53 and 84 months after third, second and first hepatectomy respectively. The median survivals from the second and first hepatic resections were 17 and 31 months, respectively. It is concluded that in the well-trained team, RHR is feasible and safe even after major primary hepatectomy. However the oncologic benefit remains questionable. PMID- 8405481 TI - The role of endoscopic ultrasonography in diagnosis of benign lesions of the upper GI tract. AB - The contribution of endoscopic ultrasonography (EUS) in the diagnosis of benign lesions of the upper gastrointestinal (UGI) tract was studied in 46 patients with polypoid lesions which were not thought to have invaded adjacent tissues or organs. Precise visualization of the five layers of the UGI tract was possible with EUS. The site of origin of the tumour was also determined. The diameter of the hypochoic lesions ranged between 1-4 cm, with well-defined margins, and were demarcated precisely from adjacent tissues. Lesions included: (1) leiomyoma of the esophagus, stomach, duodenum (n = 7); (2) ectopic pancreatic tissues (n = 3); (3) polyp of the esophagus, stomach or duodenum (n = 12); (4) hypertrophic gastric folds (n = 19); (5) extra gastric compression (n = 5). Patients from groups 1, 2 and 3 had the diagnosis confirmed and staged surgically. It was possible to differentiate between polypoid lesions of the UGI tract and extra gastric compression. The sensitivity of EUS in the diagnosis of benign tumours was 92%. PMID- 8405482 TI - Bioreduction of RSU 1069 and mitomycin C after dearterialization. AB - RSU 1069 (1-(2-nitro-1-imidazolyl)-3-(1-aziridinyl)-2-propanol) and Mitomycin C (MMC) have both been shown to be directly cytotoxic to hypoxic cells. Repeat and transient dearterializations of a liver tumour would cause its cells to become intermittently hypoxic. In this experiment the therapeutic gain of RSU 1069 and MMC was evaluated when combined with either a single or repeat dearterializations of a transplanted liver tumour. The tumour growth during the first 6 days was significantly delayed when the administration of RSU 1069 (40 mg kg-1, i.p.) was followed 15 minutes later by a single dearterialization for 2 hours compared with sham operation (P = 0.0369) or either treatment alone (P = 0.0142 vs RSU 1069 alone and P = 0.0031 vs a single dearterialization for 2 hours alone). Furthermore, RSU 169 administered 15 min prior to a single dearterialization for 2 hours is more effective to retard tumour growth than MMC either alone (P = 0.0198) or with the same period of dearterialization (P = 0.0326). Repeat dearterializations (2 hours/day) during 5 days effectively retarded the growth of the tumour by itself (P = 0.015 vs sham operation). A larger growth delay was obtained when RSU 1069 was administered 15 min before the first dearterialization (P = 0.009 vs sham operation), though the growth delay between those two groups was not significant (P = 0.07). Survival time was significantly prolonged when repeat dearterializations were combined with RSU 1069 (P = 0.007 vs sham operation; P = 0.0009 vs RSU 1069 and P = 0.003 vs repeat dearterialization alone).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405483 TI - Development of gastric cancer with special reference to growth and DNA content. Slow and rapid growing types. AB - We treated two patients with gastric cancer for whom surgery was delayed and the natural course of the tumor could be observed pre and postoperatively. A 70-year old man with a well differentiated adenocarcinoma in the upper portion of the stomach underwent resection 6 years after the clinical diagnosis. Despite extension upwards towards the lower esophagus during a 6-year period, the tumor could be resected. In a poorly differentiated adenocarcinoma in the lower third of the stomach in a 40-year-old woman, the primary lesion was not distinct on the upper GI series done 6 months before, while at the time of laparotomy, disseminating metastases and pancreas invasion made it impossible to resect the stomach together with the tumor. The tumor in the man had a low DNA ploidy, while the lesion in the woman was characterized by a high ploidy. These findings show the distinct difference in the biological behavior of the tumor in these two patients. The possible correlation between DNA content with speed of growth warrants further attention. PMID- 8405484 TI - Perianal extramammary Paget's disease. Report of two cases. AB - Perianal (extramammary) Paget's disease is rare and corresponds to an intraepithelial adenocarcinoma arising from dermal apocrine sweat glands. Biopsy reveals the diagnosis. The condition is often associated with an underlying malignancy (carcinoma of the apocrine or eccrine glands, rectal carcinoma, anal carcinoma). Wide local excision is recommended when invasive growth is absent. For invasive cancer or when associated with a synchronous malignancy abdomino perineal resection is the treatment of choice. Two cases of perianal Paget's disease are presented. PMID- 8405485 TI - Abdominal ultrasound can be helpful in diagnosis of obscure small bowel haemorrhage. AB - A case of severe gastrointestinal bleeding occurring from a rare, pedunculated and ulcerated small bowel leiomyoma is reported. The possibilities of preoperative diagnosis in an intermittent gastrointestinal bleeding case, including abdominal ultrasound and small bowel meal are discussed. PMID- 8405486 TI - Malignant fibrous histiocytoma arising in a previous site of fracture and osteomyelitis. AB - Malignant fibrous histiocytomas (MFH) are known to arise in benign pre-existing conditions. We present here a case where MFH occurred more than 40 years after a fracture and infection. The diagnosis was established only after repeated clinical and histopathologic examinations. DNA flow cytometric and chromosomal analyses were also done. PMID- 8405487 TI - Isolated splenic metastases from colon carcinoma. AB - It is generally accepted that splenic metastases from colorectal carcinoma occur concurrently with widespread dissemination of the tumor. The case reviewed is notable for its long disease-free interval, isolated splenic metastasis and favorable-outcome postsplenectomy. It represents the fourth case report in the English literature of surgical resection for an isolated splenic metastasis from colorectal carcinoma. PMID- 8405488 TI - Correlation of long-term results of extra-anatomic bypass and flow waveform analysis. AB - To investigate the correlation between the long-term results of an extra-anatomic bypass and operative flow waveform analysis, we retrospectively reviewed 32 axillofemoral and 21 femorofemoral bypasses for between 1 and 5 years after surgical repair. For the grafts with a type O or I flow waveform pattern, the patency at 3 years (83%) and at 5 years (83%) was superior to grafts with a type II flow waveform (69 and 60%). For the axillofemoral bypass grafts with a type O or I waveform, the patency rate was 92% at 5 years. In contrast with the type II flow, the patency rate was 70% at 5 years. For the femorofemoral bypass with a type O or I and a II flow waveform, the patency at 4 years was 67 and 46%, respectively. In cases with concomitant superficial femoral artery occlusion, the flow waveform was type O or I in 23% and type II in 77%. The patency rate at 5 years was 85% for cases with a patent superficial femoral artery, and 51% for an occluded superficial femoral artery (p < 0.05). These results suggest that the long-term outcome of extra-anatomic bypass correlates with the operative flow wave analysis. In addition, good long-term results and an accurate flow wave pattern depend on the distal run-off, particularly the presence of a patent superficial femoral artery. PMID- 8405489 TI - Percutaneous angioplasty of the profunda femoris artery: a safe and effective endovascular technique. AB - The limb with an occluded superficial femoral artery (SFA) relies on the profunda collaterals for adequate perfusion. Frequently the profunda is also diseased exacerbating the limb ischaemia. We have used percutaneous transluminal angioplasty (PTA) of the profunda increasingly in recent years to treat such patients. In 28 limbs there was one technical failure, no major complications and six minor wound haematomas. A combination of SFA and profunda PTA was used in 11 patients. Ten of these became asymptomatic and one improved. In 16 patients a long SFA occlusion was unsuitable for PTA. An iliac and profunda PTA was performed in six of these, with resolution of (three) or significant improvement in (three) symptoms. Profunda PTA alone was used in the remaining 10 patients who constituted a high risk elderly group all with limb threatening or disabling ischaemia. Symptomatic improvement in seven of these enabled surgery to be avoided. A bypass procedure was performed in the three patients who failed to improve. Poor run-off (< one healthy calf vessel) was the major factor limiting the effectiveness of profunda PTA in these patients. Profunda PTA is a safe and effective procedure of particular value in high risk patients with a long SFA occlusion and at least one healthy calf vessel run-off. PMID- 8405490 TI - Effects of intermittent pneumatic compression of the foot on the microcirculatory function in arterial disease. AB - OBJECTIVE: The venous pump of the foot assists blood returning to the heart. The aim of this study was to evaluate the effect of mechanical activation of the foot pump on the microcirculation of the skin in patients with peripheral occlusive arterial disease. DESIGN: Single parallel group comparing patients with arterial disease to normal control subjects. SETTING: Department of Surgery, the University College and Middlesex Hospital, London, U.K. SUBJECTS AND MATERIALS: 15 patients with peripheral occlusive arterial disease and 15 control subjects. A pneumatic impulse foot pump was applied to the foot. OUTCOME MEASURES: The Laser Doppler flux (LDF) and transcutaneous oxygen tension (tcPO2) were measured on the big toe with the subject supine, before, during and after a 10 min period of foot pumping. The study was repeated with the subject sitting. RESULTS: On sitting there is a fall in LDF and rise in tcPO2. Application of intermittent pneumatic compression of the foot in the sitting position resulted in an increase in LDF. In patients, the median percentage increase was 57% and the median difference was 82 arbitrary units (AU) (95% CI 60-130, p < 0.001). In controls, the median percentage increase was 66% and the median difference was 124 AU (95% CI 73-275 p < 0.001). There was a corresponding "further" increase in tcPO2 in both groups of subjects. In patients, the median percentage increase was 8%, in controls the median percentage increase was 10% p < 0.01). CONCLUSION: We conclude that intermittent pneumatic compression of the foot in the dependent position increases LDF and tcPO2. PMID- 8405491 TI - Experimental assessment of thrombogenicity in vascular prostheses before and during prostaglandin E1 treatment. AB - The effect of prostaglandin E1 (PGE1; Prostavasin), a powerful platelet blocking agent, was assessed on various new synthetic or biological prostheses in a 6-min in vivo extra corporal arterio-venous (AV) shunt. In eight anaesthetised and heparinised minipigs (weight 25.1 +/- 1.9 kg) the following materials were tested before and during PGE1 infusion (alprostadil-alpha-CD; 40 micrograms/50 ml NaCl/50 min or 0.8 microgram/min): PTFE, (Gore-tex, TW, 4 mm ID); Xenograft, Biologic (Solcograft, 5 mm ID), (non-porous) and Dacron (Atrium, 4 mm ID); polyurethane 1 (Braun-Melsungen, 4 mm ID); polyurethane 2 (S. Gogolewski, 4 mm ID), (porous). Technetium-labelled platelet deposition and blood flow were measured; morphology was assessed by scanning electron microscopy (SEM) and histology. Compared to control levels, PGE1 infusion had a significant systemic effect on mean arterial pressure required by the protocol (MAP 82.6 vs. 66.6 mm Hg; p < 0.001) and flow (173.6 vs. 134.8 mm/min; p < 0.001). Standardised platelet counts per area showed a marked overall decrease from 197 to 130 counts/min/mm2; NS). The morphological assessment by SEM showed a slight increase of surface cellular deposition (score: 6.7 vs. 8.3), the histology score being unchanged (3.9 vs. 3.7). Looking at deposition of platelets for each prosthesis, the porous materials showed a net improvement after PGE1 treatment as compared to non-porous materials. We conclude that PGE1 may be of benefit by reducing platelet deposition in synthetic porous vascular prostheses in the early phase. PMID- 8405492 TI - Low ankle-brachial pressure index in 68-year-old men: prevalence, risk factors and prognosis. Results from prospective population study "Men born in 1914", Malmo, Sweden. AB - The objectives were to study the distribution of ankle-brachial pressure indices (ABPI) in elderly men in relation to arteriosclerotic risk factors, cardiovascular morbidity and total mortality. The data are taken from the prospective cohort study "Men born in 1914" in Malmo, a city in southern Sweden with 220,000 inhabitants and a single referral hospital. Prevalence of low ABPI (< 0.90) at 68 years of age, total mortality, mortality from ischaemic heart disease (IHD) International Classification of Diseases (ICD) code 410.0-412.9) and cardiac event rate [fatal and non-fatal myocardial infarction (MI) and mortality from chronic ischaemic heart disease] during 8 years of follow-up was measured. Sixty-seven of 477 randomly selected men (14.0%) had an ABPI < 0.90 in one or both legs, 18/477 (3.8%) had intermittent claudication according to the Rose questionnaire, which had a sensitivity of 14.9% and a specificity of 98% when using ABPI as a reference method. Ninety-two per cent of the men with an ABPI < 0.90 were or had been smokers, compared with 80% of the men with an ABPI > or = 0.90. Mean systolic blood pressure and median plasma triglyceride levels were significantly higher in the group with low ABPI. Thirty (45%) of the men with low ABPI and 87 (21%) of the men with pressure indices above 0.90 died during follow-up (p < 0.001). Cardiac event rate was 25% (17/67) in the group with low ABPI and 10% (41/410) in the other group (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405493 TI - Intraoperative catheter thrombolysis as an adjunct to surgical revascularisation for infrainguinal limb-threatening ischaemia. AB - The objective of this study was to assess the benefits of intraoperative thrombolysis (IOL) on patients with acute leg ischaemia. This study was conducted in the Department of Cardiovascular Surgery, Inselspital, Berne, Switzerland. IOL was prospectively assessed in 25 patients with infrainguinal limb-threatening ischaemia due to acute thrombosis of atherosclerotic lesions and aneurysms (44%), occluded grafts (32%), arterial injuries (12%), delayed embolism (8%) and trash foot (8%). Three hundred and seventy-five thousand units of urokinase were delivered over 30 min with inflow occlusion to the profunda femoral artery in 8%, to the calf arteries via exposed trifurcation in 88% and to the pedal arch via exposed posterior tibial artery at the ankle in 8% of the patients. This was followed by graft thrombectomy in 24%, femoropopliteal bypass in 60%, intraoperative percutaneous transluminal angioplasty in 12% and vein patch angioplasty in 16%. Chief outcome measures were: postoperative morbidity; mortality; patency and limb salvage up to a maximum of 2 years. Postoperative bleeding complications occurred in two patients (8%) and consisted of two wound haematomas. Four patients died within 30 days after IOL, but no death could be attributed to IOL. All remaining patients were followed with a mean follow-up time of 10.9 months. The patency and limb salvage rate remained stable at 71 and 86% after 6 and 2 months, respectively. Conclusions were that IOL followed by surgical inflow restoration is a straightforward procedure for limb-threatening ischaemia with rewarding results regarding side effects, patency and limb salvage. PMID- 8405494 TI - Duplex ultrasound in patients with suspected aorto-iliac occlusive disease. AB - Duplex ultrasound scanning was used to localise and classify aorto-iliac occlusive disease. The study included 76 consecutive examinations of 73 patients with signs suggestive of proximal occlusive disease either by history or from traditional non-invasive laboratory investigation. Duplex ultrasound scanning indicated the presence of significant proximal occlusive disease in 70/101 limbs with suspected aorto-iliac disease. In total, 383/393 proximal arterial segments were assessed. A complete evaluation of the aorto-iliac region was possible in 91% of the patients. Duplex scanning was superior to oscillometric amplitude measurements and to CW Doppler examination, especially in patients with concomitant disease of the proximal superficial and deep femoral arteries. Duplex classification of stenoses correlated well with angiographic results obtained in 60 limbs with exact agreement in 194/211 (92%) arterial segments. Three of the patients with disparity between ultrasonography and angiography were investigated with intraarterial pressure measurements demonstrating that these lesions were underestimated by angiography. We conclude that Duplex ultrasound is feasible and accurate in detecting and grading lesions in the aorto-iliac region. This method provides important clinically useful haemodynamic information non-invasively in patients with suspected aorto-iliac occlusive disease. PMID- 8405495 TI - The effect of surgical preparation and in-vitro distension on the intrinsic fibrinolytic activity of human saphenous vein. AB - The objective of this study was to assess the effect of distension on the intrinsic fibrinolytic activity of human saphenous vein during its preparation for use as a bypass conduit prior to coronary artery surgery. Fibrinolytic activity was studied using fibrin plate techniques and Chromogenic assays for extractable tPA and uPA. Samples were obtained from patients undergoing routine coronary surgery. Fibrinolytic activity was compared in control vein (untouched) vein that had been prepared for use as a bypass conduit (surgical dissection, ligation of side branches and careful but uncontrolled manual distension) and segments of dissected vein distended in vitro to pressures of 230 or 120 mmHg. Uncontrolled distension and distension to 230 mmHg impaired fibrinolytic activity as determined by areas of lysis on fibrin plates (p < 0.05), (p < 0.005), tPA activity (p < 0.005) (p < 0.005) and uPA activity (p < 0.05), (p < 0.005). Distension to 120 mmHg had no effect on the fibrinolytic activity of human saphenous vein. Impaired fibrinolytic activity caused by uncontrolled distension of saphenous vein prior to its use as a vascular conduit may contribute to early vein graft thrombosis and can be avoided using controlled distension to < 120 mmHg. PMID- 8405496 TI - Prevalence of intermittent claudication and risk factors for its development in patients on renal replacement therapy. AB - The prevalence of symptomatic intermittent claudication (IC) was assessed using a standard cardiovascular questionnaire in a cohort of 325 patients on renal replacement therapy (RRT). IC was found in 19% of patients, 77% of whom were smokers and 22% diabetic. It was more common in men than women and in smokers than non-smokers (p < 0.001). Those with IC were significantly older (61 years vs. 50 years p < 0.001), smoked more (23 pack years vs. 12 pack years p = 0.002), had higher median systolic blood pressures (143 mmHg vs. 140 mmHg p = 0.041) and median triglyceride levels (2.07 mmol/l vs. 1.60 mmol/l p = 0.023) than those renal patients without IC. A case control study matching for age, sex and treatment revealed patients with IC to have higher median systolic blood pressure (147 mmHg vs. 140 mmHg p = 0.031), cholesterol (6.70 mmol/l vs. 5.90 mmol/l p = 0.029), LDL cholesterol (4.64 mmol/l vs. 3.86 mmol/l p = 0.011), and contained a greater proportion of smokers (78% vs. 50% p < 0.001). IC is common in patients on RRT. Whilst smoking was prevalent among those with IC it was much less frequent than in the general population with IC. Other factors such as hypertension, lipid abnormalities or the uraemic state itself may also be important in the development of IC in these patients. PMID- 8405497 TI - ePTFE grafts for femoro-crural bypass--improved results with combined adjuvant venous cuff and arteriovenous fistula? AB - Patency rates for long prosthetic bypass grafts with standard anastomoses to single tibial or peroneal arteries are very poor. Adjuvant techniques employed with the aim of improving patency rates include arteriovenous fistula (AVF) at the distal anastomosis to accelerate blood flow above thrombotic threshold velocity (TTV) and a venous cuff (VC) or patch which may reduce or modify anastomotic myointimal hyperplasia within the recipient artery. In a consecutive series of 43 femoro-crural bypasses with ePTFE grafts, adjuvant AVF and VC procedures have been applied in combination. The results are compared with those of an antecedent series of 76 similar grafts with AVF alone and a contemporaneous series of 179 autologous vein grafts. All operations were undertaken for critical limb ischaemia with anastomosis to a single calf or pedal artery. The three groups were well matched for age, sex, diabetes, smoking history, previous surgery and the proportion with rest pain and tissue necrosis. The cumulative patency rate at 2 years for ePTFE grafts with combined AVF and VC was 62% compared to 28% for those with AVF alone and 68% for autologous vein grafts. The patency rate for prosthetic grafts with AVF and VC was significantly higher than AVF alone (p < 0.01) and did not differ significantly from vein grafts. Cumulative limb salvage rates for ePTFE grafts with AVF and VC were 68% at 1 year and 55% at 2 years compared to 38 and 35% for AVF alone and 78 and 69% for vein grafts.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405498 TI - Endotoxaemia, the generation of the cytokines and their relationship to intramucosal acidosis of the sigmoid colon in elective abdominal aortic aneurysm repair. AB - Ischaemia of the large bowel occasionally occurs following abdominal aortic aneurysm repair and may lead to multiple system organ failure (MSOF). Intramucosal acidosis of the sigmoid colon is a good indicator of sigmoid colonic ischaemia. Intramucosal pH of the sigmoid colon was measured using the silicone tonometer in 21 patients undergoing abdominal aortic aneurysmectomy. Samples were taken for plasma endotoxin, tumour necrosis factor (TNF) and interleukin-6 (IL-6) measurements preoperatively, half-hourly during the operation, 2-hourly for the next 12 h, 4-hourly for a further 48 h and 8-hourly thereafter until the fifth day. The intramucosal pH of the sigmoid colon fell to less than 7.00 peri operatively in 10 patients, four of whom developed diarrhoea; in comparison, this did not occur in any of the 11 whose pH remained greater than 7.00 (p = 0.036). Higher peak concentrations of endotoxin, TNF and IL-6 were found in those patients whose intramucosal pH fell to less than 7.00 compared to those whose pH remained greater than 7.00 (mean +/- S.E.M. pg/ml, endotoxin = 112 +/- 24 vs. 58 +/- 6, p < 0.05; TNF = 26 +/- 8 vs. 7 +/- 2, p < 0.05; IL-6 = 213 +/- 59 vs. 87 +/- 12, p = 0.09). In the two patients who died, both from the group with pH level less than 7.00, concentrations of IL-6 were considerably higher than that in most of the other patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405499 TI - Lipid peroxidation as a cause of lower limb swelling following femoro-popliteal bypass grafting. AB - We examined the role of free radical induced lipid peroxidation in lower limb swelling in patients following femoro-popliteal bypass grafting. In 20 patients undergoing this operation blood samples were taken from the femoral vein via a cannula before the femoral artery clamp was applied, just prior to and immediately after clamp release and at 10 min intervals thereafter for 1 h for measurements of malondialdehyde (MDA) and vitamin E. The concentration of MDA was significantly elevated at 40 min after reperfusion (mean +/- S.E.M., 573 +/- 83 pmol/ml) compared to just before clamp release (359 +/- 41 pmol/ml; p < 0.01). This was associated with a corresponding fall in the concentration of vitamin E at the time of peak MDA rise (5.68 +/- 0.28 to 5.29 +/- 0.28 mumol/mM cholesterol, p < 0.05) suggesting its utilisation as an antioxidant. The degree of oedema was related to the changes in MDA and vitamin E. Thus, in the 15 patients with greater than 10% increase in lower limb volume the rise in the concentration of MDA was 364 +/- 44 to 693 +/- 76 pmol/ml (p = 0.0001) while that in the five, whose swelling was less than 10%, was 344 +/- 40 to 559 +/- 243 pmol/ml (p = 0.25). A significant fall in vitamin E was found only in the group with greater than 10% lower limb oedema (5.90 +/- 0.33 to 5.40 +/- 0.34 mumol/mM cholesterol, p < 0.01), in comparison to those with less than 10% swelling (5.01 +/- 0.35 to 5.04 +/- 0.50 mumol/mM cholesterol).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405500 TI - Content and turnover of extracellular matrix protein in human "nonspecific" and inflammatory abdominal aortic aneurysms. AB - Inflammatory aneurysms (IAs) have peculiar macroscopic and histological aspects which make them very different from nonspecific aneurysms (NSAs). These morphological differences seem to be determined by significant modifications of the extracellular matrix. Extracellular matrix protein component concentrations were determined biochemically in infrarenal aortic biopsies from 10 NSAs, five IAs and five non-aneurysmal aortic controls. The concentration of each wall component was expressed in % w/w (relative concentration) and in mg/wall longitudinal cm (absolute concentration) with reference to total protein recovered after hydrolysis and amino acid analysis. The biochemical results were correlated with the histological and ultrastructural features of the specimens. A significant increase in total collagen was observed in the two groups of aneurysms, with respect to the controls (NSA = 285%, IA = 382%). In contrast the 80-90% decrease in the relative concentration of elastin observed in both types of aneurysm was less marked (NSA = 55%, IA = 39%). This fall was not significant when expressed in mg/cm, although elastin derived peptide (EDP) levels in the plasma of these patients was significantly higher than in age-matched controls. The concentration of the soluble collagen fraction appeared significantly higher (Mann-Whitney, p < 0.05) in the IAs with respect to the NSAs, whilst no differences were observed between the two groups regarding the concentration of insoluble elastin and of wall and plasma EDPs. As well as providing evidence of increased elastin turnover, this study emphasises the conspicuous modifications of collagen deposition in the wall of abdominal aortic aneurysms which appeared more marked in the inflammatory group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405501 TI - A randomised controlled trial of a low-molecular-weight heparin (Enoxaparin) to prevent deep-vein thrombosis in patients undergoing vascular surgery. AB - The incidence of postoperative deep vein thrombosis (PDVT) after aortic surgery and lower limb revascularisation has not been assessed by a large prospective study. In a prospective randomised trial the effect of a low-molecular-weight heparin fragment, Enoxaparin (ENX) 4200 anti factor Xa IU once daily was compared to that of unfractionated heparin (UFH) 7500 IU twice daily. Two hundred and thirty-three consecutive patients were classified into three groups, aortic or aortoiliac and aneurysmectomy (n = 75), aorto-femoral bypass for atherosclerotic disease (n = 71), and femoropopliteal or femorodistal bypass (n = 87). Patients were analysed for development of deep vein thrombosis by Duplex scanning and, if positive, by venography between the seventh and tenth postoperative day. PDVT was present in 10 patients in the ENX group and in four patients in the UFH group (8.2 and 3.6% respectively, NS). The incidence of PDVT was 8% after aortic or aortoiliac aneurysmectomy, 7% after aortofemoral revascularisation, and 3.4% after femoropopliteal or femorodistal bypass. The overall incidence of PDVT after aortic surgery was 7.5% (95% CI 5.4-9.7). There was no pulmonary embolism. Intra operative blood loss and postoperative bleeding events did not differ significantly between the ENX and UFH groups. After 1 month follow-up, no clinical event or death could be related to PDVT or pulmonary embolism. In conclusion, in vascular surgery ENX is as safe and effective in the prevention of PDVT as is UFH. PMID- 8405502 TI - Abdominal aortic aneurysm: ten years' hospital population study in the city of Glasgow. AB - The study comprised 2936 cases of aortic aneurysm admitted to Glasgow City hospitals between January 1980 and December 1989. Information was first obtained from Scottish Morbidity Records 1 (SMR1), Scottish Hospital Inpatients Statistics, the accuracy of which was assessed by detailed inspection of 500 case notes, evenly distributed throughout the study period. The data from 489 of the 500 case notes examined matched the SMR1 data which was therefore accurate in 97.8% of cases. Of the 2936 cases, 852 (29%) had more than one admission with the same diagnosis in the same year and of the remaining 2084 cases 169 (8.1%) were thoracic aneurysms. After correction for these findings, 1915 abdominal aortic aneurysms (AAA) were left for the study. Of these, 618 (32.3%) were females giving a male to female approximate ratio of 2:1 which remained constant for each year of the study. The mean age of the study population increased from 70.3 years in 1980 to 72.3 years in 1989 with an overall mean of 71.2 years. There was a significant increase in the percentage of patients over the age of 75 years (from 17.4% in 1980 to 28.8% in 1989). Twenty-seven per cent of the cases (519 of 1915) were ruptured aneurysms. This was again stable for the 10 years of the study. Analysis of hospital mortality suggested better outcomes, both for ruptured and non-ruptured aneurysms, in the purely vascular unit of Glasgow as opposed to other surgical units in the city. The possible significance of this finding is discussed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405503 TI - Infragenicular in situ vein bypass graft occlusion: a multivariate risk factor analysis. AB - Early postoperative thrombosis and the later development of graft stenoses are the two major causes of vein bypass graft failure. The risk factors for both these outcomes were analysed in a multivariate analysis of 82 consecutive infragenicular in situ vein grafts. Twenty-four grafts failed within 30 days but eight were successfully revised. Technical errors accounted for six of the failures. A multivariate analysis revealed graft resistance > 1.4 peripheral resistance units (odds ratio 5.8, 95% C.I. 1.6-20) as the only independent risk factor for early graft failure. Eighteen grafts (27%) developed a stenosis most commonly in the distal third of the graft (46%). Poor quality, small diameter vein was the only independent risk factor for graft stenosis (odds ratio 7, 95% C.I. 1.5-34). Composite vein grafts, where narrowed and thickened vein had been replaced, had a significantly lower stenosis rate (difference in proportions 0.41, 95% C.I. 0.1-0.8, Mann-Whitney U test). PMID- 8405504 TI - The influence of smoking and lipids on restenosis after carotid endarterectomy. AB - Factors associated with restenosis were investigated in 107 patients undergoing carotid endarterectomy for symptomatic disease. The patients, 71 men and 36 women with mean age 68 +/- 8 years, were followed up for 1 year by serial Duplex scanning. Carotid restenosis of > or = 50% developed in 18 patients (17%), 11 men and seven women. Restenosis was not influenced by age, sex, diabetes or hypertension. Continuing smokers, serum cotinine > 200 nmol/l, had a significantly higher incidence of > or = 50% restenosis after 1 year (39%), compared with only 16% of non-smokers, p = 0.023. Restenosis > or = 50% also was associated significantly with below median body mass index (p = 0.027). Women undergoing carotid endarterectomy had higher levels of cholesterol (median 7.4 mmol/l) and apolipoprotein B (median 0.81 g/l) than men (median cholesterol 6.4 mmol/l, median apolipoprotein B 0.69 g/l), p < 0.01. For men only, restenosis of > or = 50% was associated with low levels of serum cholesterol (median 5.7 mmol/l), p = 0.002. For women cholesterol levels were higher (median 8.1 mmol/l) in those with > or = 50% restenosis. Smoking adversely influences early restenosis (1 year) after carotid endarterectomy. Hyperlipidaemia is not a risk factor for restenosis in men, but may be associated with restenosis in women. PMID- 8405505 TI - The surgical management of high-output cardiac failure due to arteriovenous haemodialysis fistulae. PMID- 8405506 TI - Septic arthritis of the knee: complication of intra-arterial thrombolytic therapy. AB - A case of septic arthritis of the knee complicating intra-arterial thrombolysis is reported. Septic complications following the use of intravascular catheters are usually relatively minor. This serious case and the discussion that arises highlights the need for shortening the duration of intravascular thrombolysis regimes. PMID- 8405507 TI - Acute abdominal aorta embolism caused by primary cardiac echinococcus cyst. AB - Embolism of the abdominal aorta by an echinococcus cyst is extremely rare and is due to rupture of an intracardiac hydatid cyst. We report a case of abdominal aortic embolism by a primary intracardiac echinococcus cyst which was treated successfully with bilateral femoral embolectomy followed by direct aortotomy. We found only 16 previous cases reported and only one caused by a primary cyst. PMID- 8405508 TI - A case of ruptured abdominal aortic aneurysm in association with congenital kyphoscoliosis. AB - We present the case of a 36-year-old woman with severe kyphoscoliosis in whom the diagnosis of ruptured abdominal aortic aneurysm was not made preoperatively and who died in the operating theatre. We think the presence of an abdominal aortic aneurysm in this relatively young patient was resultant upon irregular flow patterns in the grossly deformed aorta. PMID- 8405509 TI - Mycotic aneurysms of the carotid artery: ligation vs. reconstruction--case report and review of the literature. AB - Mycotic aneurysms of the extracranial carotid artery are very uncommon, with only twenty-nine cases reported in the medical literature. This is a report of a mycotic aneurysm of the common carotid artery caused by septic cervical lymphadenopathy. A review of the literature is presented in an attempt to elucidate which surgical approach is superior, ligation or repair of the carotid artery. PMID- 8405510 TI - Salpingitis isthmica nodosa: a review of the literature, discussion of clinical significance, and consideration of patient management. AB - OBJECTIVES: To examine and discuss the pathology, diagnosis, incidence, and patient profile of salpingitis isthmica nodosa and to question its natural history, propose management strategies, and identify areas of promising research. DESIGN: Over 50 studies were reviewed, evaluated, and compared to offer the clinician a foundation on which to generate treatment plans. RESULTS: Salpingitis isthmica nodosa is diagnosed by the pathological presence of isthmic diverticula and may be suggested by characteristic changes on hysterosalpingogram. Its incidence in healthy, fertile women ranges from 0.6% to 11%, but it is significantly more common in the setting of ectopic pregnancy and infertility. There are no studies, retrospective or prospective, that clearly dictate appropriate therapy. CONCLUSION: Given its progressive nature and probable deleterious effects on fertility, we propose that microtubal surgery be the definitive treatment for qualified women who have salpingitis isthmica nodosa. PMID- 8405511 TI - Menopausal health is scientifically and clinically important. PMID- 8405512 TI - Sequential regimen of the antiprogesterone RU486 and synthetic progestin for contraception. AB - OBJECTIVE: To examine the effect of sequential use of the antiprogesterone RU486 and synthetic progestin on ovarian function of healthy women. DESIGN: Healthy women were given a sequential antiprogesterone-progestin treatment. Blood samples were taken twice a week during one control cycle and one to three treatment cycles; prospective analysis. SETTING: The outpatient clinic of the Helsinki City Maternity Hospital, Helsinki, Finland, and Steroid Research Laboratory, Department of Medical Chemistry, University of Helsinki, Helsinki, Finland. PATIENTS: Eleven healthy women, volunteers, 20 to 34 years of age. INTERVENTIONS: A dose of 25 mg/d of RU486 was given during cycle days 1 to 21, and synthetic progestin (5 mg of norethisterone to six and 5 mg of medroxyprogesterone acetate to five women) during cycle days 22 to 31. MAIN OUTCOME MEASURES: Serum P, E2, FSH, and LH were measured from serum samples. RESULTS: In 20 of the 24 treatment cycles analyzed the serum concentrations of P were anovulatory. In the remaining 4 cycles, P levels rose above 3 ng/mL, suggestive of ovulation. Folliculogenesis was not completely inhibited, but serum E2 profiles were subnormal and delayed. Bleeding control was satisfactory. CONCLUSIONS: Antiprogesterone RU486 hampers or delays follicular development, suggesting a possible use as an estrogen-free oral contraceptive. However, the synthetic progestins used in this regimen induced serum P rises in some cycles. The synthetic progestin provides the cycle control, but its possible effect on the reliability of the method remains to be evaluated. PMID- 8405513 TI - Ovulation inhibition with a combined oral contraceptive containing 1 mg micronized 17 beta-estradiol. AB - OBJECTIVE: To evaluate ovarian function by ultrasonography and endocrine measurements. DESIGN: Prospective, open study. SETTING: Outpatient clinic of the First Department of Gynaecology and Obstetrics, University of Vienna, Austria. PARTICIPANTS: Twenty healthy women with regular cycles and established ovulation by ultrasonography. INTERVENTION: Treatment with a combination of 1 mg micronized E2 with 150 micrograms desogestrel daily for 21 days, followed by 7 pill-free days. MAIN OUTCOME MEASURES: Transvaginal ultrasonography and estimation of E2 and P at least twice a week for two consecutive cycles, followed by one after treatment cycle. RESULTS: Ovulation inhibition was apparent in all cases and no functional ovarian cysts were observed during treatment. On a few occasions a persistent follicle was noted, but in the majority of cases there was total absence of follicular activity. The bleeding pattern showed a tendency toward prolonged and more heavy bleeding when compared with the before treatment situation. Return of ovulation was prompt in all women but one. CONCLUSIONS: Ultrasonographic observations, accompanied by P and E2 measurements, allow us to conclude that the combination of 1 mg E2 with 150 micrograms desogestrel provides complete ovulation inhibition. However, the bleeding pattern does not show an acceptable profile. PMID- 8405514 TI - The relationship between follicle-stimulating hormone dose and level and its relevance for ovulation induction with adjuvant gonadotropin-releasing hormone agonist treatment. AB - OBJECTIVE: To investigate the effect of a GnRH agonist (GnRH-a) on the FSH threshold level and the relationship between the FSH dose and the FSH level of patients suffering from polycystic ovarian syndrome (PCOS). DESIGN: The stimulation with low-dose FSH in PCOS (group 1) was compared with the subsequently performed stimulation with low-dose FSH combined with GnRH-a in another group of patients suffering from the same syndrome (group 2). SETTING: Specialist Reproductive Endocrine Unit. PATIENTS: Suffering from clomiphene citrate-resistant PCOS. MAIN OUTCOME MEASURES: The FSH threshold level for ongoing follicular growth and the relationship between dose and level of FSH. RESULTS: In 15 patients in group 1 and in 13 patients in group 2, respectively, 39 and 32 stimulation cycles were performed. Below and above threshold values of FSH of group 1 and 2 did not differ significantly. For the equation stable level of FSH (Y mIU/mL) = A X infusion rate of FSH (X IU/24 h) + basal level of FSH (B mIU/mL), the median A of group 1 was 0.027 and A of group 2 was 0.055 (significant difference). CONCLUSIONS: In PCOS, a change of the FSH threshold level for ongoing follicular growth induced by the GnRH-a could neither be proven nor ruled out. The use of a GnRH-a resulted in larger FSH level increases per IU/24 h of FSH administered and might therefore interfere with the effect of low dose FSH treatment. PMID- 8405515 TI - The influence of luteinizing hormone and insulin on sex steroids and sex hormone binding globulin in the polycystic ovarian syndrome. AB - OBJECTIVE: To examine the relationship between hyperinsulinemia, sex hormone binding globulin (SHBG), and body mass index (BMI) on LH-induced hyperandrogenemia in patients with polycystic ovarian syndrome (PCOS). DESIGN: Insulin responses during an oral glucose tolerance test (OGTT) were assessed in 25 consecutive women with PCOS and 20 control women matched for BMI. Insulin responses and sensitivity (SI) were also determined using a frequently sampled intravenous glucose tolerance test (IVGTT). SETTING: The clinical research center at a university medical center. MAIN OUTCOME MEASURES: Serum LH, SI, and basal, peak, and area under the curve (AUC-insulin responses) were determined and correlated with SHBG, androstenedione (A), T, and free T concentrations. RESULTS: Compared with controls, the AUC-insulin response during OGTT was greater in PCOS, with an average increase of 44%. During IVGTT, AUC-insulin response was also significantly higher in PCOS versus controls, with an average increase of 53%. In addition, SI was reduced in PCOS versus controls with an average decrease of 53%. The average differences in oral- and intravenous-glucose-induced hyperinsulinemia and in insulin sensitivity between PCOS and controls were relatively constant across the entire physiological range of BMI. In PCOS, baseline LH showed strong positive correlations with baseline A and T. However, there were no significant correlations between either basal, peak, or AUC-insulin response during OGTT and IVGTT with basal T or A concentrations or between insulin and androgen levels measured at 30-minute intervals throughout the OGTT. However, basal, peak, and AUC-insulin responses during OGTT were strongly correlated with fasting SHBG binding capacity. CONCLUSIONS: These data are consistent with the hypothesis that hyperinsulinemia in PCOS influences the biologically active component of T by lowering SHBG concentrations while having little apparent impact on LH-induced secretion of androgens in vivo. PMID- 8405516 TI - Insulin resistance improves in hyperandrogenic women treated with Lupron. AB - OBJECTIVE: To examine if changes in insulin sensitivity and glucose effectiveness in women with polycystic ovarian disease (PCOD) occurred after ovarian androgen suppression with a GnRH agonist, leuprolide acetate (LA, Lupron; TAP Pharmaceuticals, Deerfield, IL) using the minimal model method. DESIGN: Twelve patients with PCOD were tested in the untreated state (baseline) and after 6 weeks of LA treatment. Subjects were divided into two groups based on the degree of impairment of their baseline insulin sensitivity index (SI; (min-1) (microU/mL 1): mild insulin resistance (SI > 1) or severe insulin resistance (SI < 1). RESULTS: In all patients, serum T was significantly decreased from elevated baseline levels to normal female concentrations after 6 weeks of LA therapy. Insulin sensitivity in PCOD patients with mild insulin resistance significantly improved from baseline after 6 weeks of LA therapy, whereas no change in SI on LA therapy was seen in PCOD women with severe insulin resistance. Glucose utilization independent of increased insulin secretion did not change as a function of LA treatment in either group. CONCLUSION: These findings indicate a significant improvement in SI in mildly insulin-resistant women with PCOD after suppression of ovarian function with LA treatment. PMID- 8405517 TI - Rapid regression of uterine leiomyomas in response to daily administration of gonadotropin-releasing hormone antagonist. AB - OBJECTIVE: The efficacy of acute and sustained pituitary gonadotropin down regulation by the Nal-Glu GnRH antagonist (Nal-Glu) was evaluated in the treatment of uterine leiomyomas. DESIGN: Prospective, open clinical trial. PATIENTS: Seven normally cycling women with symptomatic leiomyomas. INTERVENTIONS: Nal-Glu (50 micrograms/kg per day) was administered subcutaneously for 3 months. MAIN OUTCOME MEASURES: Baseline ultrasound examinations were obtained and repeated monthly throughout treatment. Each leiomyoma was mapped and measured in three dimensions. Blood samples were drawn daily for 7 days, weekly for 4 weeks, and monthly for the remaining 2 months. RESULTS: Mean leiomyoma size decreased 52.8 +/- 7.3% (means +/- SD) after 1 month of therapy and remained unchanged for the remainder of the study. Serum levels of E2 (35.9 +/- 11.8 to 9.3 +/- 0.8 pg/mL, 131.7 +/- 43.3 to 34.0 +/- 1.4 pmol/L), estrone (37.3 +/- 7.5 to 13.0 +/- 2.5 pg/mL, 138.1 +/- 27.7 to 48.1 +/- 9.1 pmol/L), and P (1.6 +/- 1.1 to 0.3 +/- 0.01 ng/mL, 5.0 +/- 3.6 to 0.9 +/- 0.04 nmol/L) declined rapidly (within 48 hours) and remained suppressed throughout treatment. Serum LH, FSH, androstenedione, T, and DHEA levels did not change significantly. In two subjects who did not have surgical removal, leiomyomas grew to original size within the 1st month off drug. Six patients remained amenorrheic and the other subject spotted during the last 2 months of therapy. CONCLUSIONS: Continuous treatment with Nal-Glu induces immediate and sustained pituitary-gonadal down-regulation that results in regression in leiomyoma size. By circumventing GnRH agonist induced pituitary-ovarian up-regulation, GnRH antagonists may prove to be superior tools in the medical management of leiomyomas. PMID- 8405518 TI - Endometrial histology after electrocoagulation using different power settings. AB - OBJECTIVE: To study endometrial histology after electrocoagulation in an in vitro model using 50 watts (W) and 100 W of coagulation current and determine the depth of endometrial destruction and survival, if any, of glands beneath this zone. DESIGN: Twenty fresh uteri of similar weights and dimensions were obtained from patients undergoing hysterectomy for benign disease. Specimens were bivalved into anterior and posterior walls and each wall divided in half. Endometrial electrocoagulation was carried out with a 5-mm probe at 50 W and 100 W applied to anterior and posterior quarters of the specimen, respectively. The adjacent untreated endometrial surfaces served as controls. Specimens were formalin-fixed, embedded in paraffin, and sections stained with hematoxylin and eosin. MAIN OUTCOME MEASURES: The number and morphology of the endometrial glands were counted and classified manually for each section and compared between each power setting and controls. RESULTS: Histologic examination revealed morphologically normal glands in all specimens beneath the zone of destruction regardless of power setting. Both power settings produced significant focal and diffuse glandular and stromal destruction when compared with controls. Significant differences were noted in the number of normal glands after treatment with 50 W (71.33 glands +/- 76.44 [mean +/- SD]), 100 W (21.11 +/- 35.71) and untreated controls (240.16 +/- 110.81). Tissue destruction increased with increasing power, and there were significant differences in the percentage of morphologically normal, surviving glands between 50 W (11.7% +/- 11.4% [mean +/- SD]) and 100 W (4.9% +/- 10.9%). CONCLUSION: These data suggest that electrocoagulation may result in a variable degree of endometrial destruction dependent on power. Viable glands and stroma may survive beneath the zone of destruction regardless of power. Such variations in endometrial insult in an in vitro model may explain, in part, the variable clinical results of endometrial electrocoagulation. The survival of glands beneath the zone of destruction in this model raises the theoretical concern for occult malignant changes and leaves open to question the exact role and mode of hormonal therapy during the menopause after endometrial ablation. PMID- 8405519 TI - Evaluation of the impact of intraobserver variability on endometrial dating and the diagnosis of luteal phase defects. AB - OBJECTIVE: To determine the magnitude of intraobserver variation in dating endometrial biopsies and its impact on clinical management. DESIGN: Blinded histopathologic interpretation of endometrial biopsy specimens 1 year apart by five pathologists. SETTING: Large military tertiary care center. PATIENTS: Endometrial biopsy specimens from 51 patients undergoing evaluation for potential luteal phase defects. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Calculation of the magnitude of the individual and overall intraobserver variation in endometrial dating for the five pathologists and estimation of its potential impact on clinical management. RESULTS: The intraobserver variation was 0.69 +/- 0.05 days (means +/- SE). There was no significant difference in the magnitude of the variation for 1-day or 2-day dating ranges. The theoretical probability of altering clinical management by having the same pathologist redate a given specimen ranged from 15% to 28%. CONCLUSION: Histologic dating of endometrial biopsies is subject to a small but highly clinically significant intraobserver variability that may have a major impact on clinical management. PMID- 8405520 TI - Immunoglobulin (Ig) G, IgA, and IgA subclass antibodies against fertilization antigen-1 in cervical secretions and sera of women of infertile couples. AB - OBJECTIVES: To assess the occurrence of immunoglobulin (Ig) G, IgA, and IgA subclass antibodies against human sperm fertilization antigen-1 (FA-1) in cervical mucus (CM) and serum of women of infertile couples. DESIGN: Enzyme linked immunosorbent assay methodology was used to detect anti-FA-1 antibodies. Antisperm antibodies were detected by agglutinating, immobilizing, and indirect immunobead (IB) methods. Control samples for the ELISA were from 10 women negative in the antisperm antibody assays. PARTICIPANTS: Samples were from women of 32 infertile couples undergoing antisperm antibody analysis. RESULTS: One of 10 control CM samples was slightly positive for IgG anti-FA-1 and none for IgA. Of the 22 CM samples from antisperm antibody-positive women, 9 were positive for IgG antibodies, 9 for IgA, 7 for IgA1, and 6 for IgA2. Cervical mucus samples from eight women were positive for both IgA and IgG antibodies. Assay of 19 serum samples, including 8 controls, by ELISA, indicated 9 of 11 from antisperm antibody-positive women and none from controls were positive for IgA and IgG (7 of 9 identical women). In addition, of the nine IgA-positive sera, seven were of the A1 subclass and five were of the A2 subclass. Positive IB assays occurred more frequently in CM and serum samples positive for anti-FA-1 antibodies than in negative samples. CONCLUSION: The results suggest that cervical secretions and sera of antisperm antibody-positive women contain IgA and IgG antibodies against sperm antigen FA-1 that may be involved in antifertility effects. PMID- 8405521 TI - A possible bimodal effect of estrogen on insulin sensitivity in postmenopausal women and the attenuating effect of added progestin. AB - OBJECTIVE: To determine the effects of estrogen and of added progestin on carbohydrate tolerance in postmenopausal women. DESIGN: An insulin tolerance test (ITT) was used to assess insulin resistance in healthy post-menopausal women and to determine the effects of oral estrogen with and without added progestin on insulin sensitivity. SETTING: A menopause research clinic at a University Medical Center. PATIENTS: Fifteen healthy postmenopausal and nine premenopausal women were studied after having not received any hormone preparations for > or = 4 weeks. INTERVENTIONS: All subjects received a baseline ITT and postmenopausal women were then randomized to receive either 0.625 mg conjugated equine estrogen, 0.625 mg conjugated equine estrogen/10 mg progestin, or 1.25 mg conjugated equine estrogen for 2 months at which time a second ITT was performed. In the former two groups the women were treated for an additional 4 months to assess the long-term effects of treatment and had a third ITT performed at the end of 6 months. MAIN OUTCOME MEASURES: Fasting serum insulin and glucose were measured and K(itt) values were obtained at each visit in each group. RESULTS: Forty-four percent of nonobese healthy postmenopausal women were found to have insulin resistance. The three groups differed significantly in their K(itt) responses. Estrogen replacement improved insulin sensitivity (K(itt) increased by 25%). However, 1.25 mg of conjugated equine estrogen caused a 24.7% decrease in K(itt) values and progestins attenuated the beneficial effects of 0.625 mg conjugated equine estrogen from baseline values (K(itt) decreased by 17.0%). Two- and 6-month values did not differ. CONCLUSIONS: Insulin resistance is prevalent in healthy postmenopausal women. A moderate dose of estrogen appears to increase insulin sensitivity but higher doses may attenuate this benefit and progestins may cause a decrease in insulin sensitivity. PMID- 8405522 TI - Determination of the steroidogenic capacity in premature ovarian failure. AB - OBJECTIVE: To determine the secretion of precursors, intermediate and final products of androgen biosynthesis in women with premature ovarian failure (POF). PATIENTS: Seven patients 20 to 34 years of age with idiopathic POF and a control group of six women 27 to 29 years of age with normal ovarian function studied during the early follicular phase were included. DESIGN, INTERVENTIONS: In all patients an adrenal stimulation test was performed as follows: a short dexamethasone (DEX) inhibition was carried out the night before the corticotropin (ACTH, 0.25 mg, Cortrosyn; Organon, Orangeburg, NY) stimulation test, obtaining blood samples in basal conditions, post-DEX inhibition as well as at 60, 120, and 180 minutes after the ACTH bolus. MAIN OUTCOME MEASURES: Using specific RIA serum concentrations of delta 5 precursors (pregnenolone, 17-hydroxypregnenolone, DHEA), delta 4 intermediates (P, 17-hydroxyprogesterone, androstenedione) and the final products T and cortisol (F) were measured. RESULTS: Adrenal inhibition and stimulation responses in both groups of patients showed no differences for delta 5 precursors and F. On the other hand, delta 4 intermediates and androgens exhibited significant differences at the level of response to ACTH stimulation. Patients with POF had significantly lower values than those of control group. CONCLUSION: An important decrement in the steroidogenesis was noticed in POF, particularly in androgen synthesis, revealing the selective participation of the adrenal gland in steroid production. PMID- 8405523 TI - Long-term gonadotropin-releasing hormone agonist with standard postmenopausal estrogen replacement failed to prevent vertebral bone loss in premenopausal women. AB - If indeed younger women do not receive bone protection from standard postmenopausal hormone replacement, these women may also be at increased risk for heart disease, urogenital atrophy, and other effects of long-term hypoestrogenism with 0.625 mg of CE add-back. We recommend that until long-term studies using BMD of the lumbar spine are available, 1.25 mg of CE or the equivalent dosage of other Es be prescribed when planning long-term GnRH-a ovarian suppression. PMID- 8405524 TI - Premature luteinization in controlled ovarian hyperstimulation has no adverse effect on oocyte and embryo quality. AB - OBJECTIVE: To determine if premature luteinization has an adverse effect on oocyte and, hence, embryo quality. DESIGN: Retrospective evaluation of anonymous ovum donors/oocyte recipients. SETTING: A large oocyte donation program. PATIENTS, PARTICIPANTS: Sixty-eight women undergoing controlled ovarian hyperstimulation (COH) as ovum donors were matched to 68 women with ovarian failure as ovum recipients who had endometrial maturation exogenously controlled by an identical hormone replacement protocol. INTERVENTIONS: Serum was collected for E2 and P in donors and recipients. MAIN OUTCOME MEASURES: The incidence of premature luteinization was determined in donors. Cycle characteristics were compared between donors with and without premature luteinization, with emphasis on oocyte and embryo quality. Implantation rates per embryo and delivery rates per transfer were measured in recipients. RESULTS: Twenty-one (31%) of the donors demonstrated premature luteinization. Serum P was higher on day before hCG, day of hCG, and day after hCG in women demonstrating premature luteinization. However, there were no differences between donor cycles with or without premature luteinization as determined by donor age, ampules of gonadotropins used, day of hCG administration, peak E2, total number of oocytes, and number of mature oocytes retrieved. Ovum recipients were of similar age and had similar E2 exposure (area under the E2 curve) before P administration. Similar fertilization rates, incidence of polyspermia, number of embryos transferred of similar embryo grade, and similar implantation rates and deliveries per transfer were observed in women receiving oocytes from donors with and without premature luteinization, respectively. CONCLUSIONS: Similar oocyte quality, fertilization, and polyspermia rates, embryo quality, implantation, and delivery rates suggest that any negative impact of premature luteinization on pregnancy rates in COH cycles from young women is not due to an adverse effect of PL on oocyte and hence embryo quality, but rather on the endometrial environment. PMID- 8405525 TI - Single-dose administration of the gonadotropin-releasing hormone antagonist, Nal Lys (antide) to healthy men. AB - OBJECTIVE: To evaluate the ability the Nal-Lys GnRH antagonist ([N-Ac-Nal (2)1, 4ClDPhe2, D3Pal3, Lys (Nic)5, D-Lys(Nic)6, Lys (iPr)8, D-Ala10] to suppress gonadotropins and T in humans and to assess its duration of action and its local effects. DESIGN: Placebo-controlled clinical study. SETTING: A university community. SUBJECTS: Seven normal male volunteers. INTERVENTIONS: We administered single injections of Nal-Lys (0, 10, 25, and 50 micrograms/kg body weight). Blood samples were collected before and at frequent time intervals after injection. RESULTS: Nal-Lys caused only minor local effects. At the higher doses (25 and 50 micrograms/kg), serum LH and T levels were suppressed to 50% to 70% of baseline; serum FSH levels were suppressed to 70% to 80% of baseline, and levels of all three hormones returned to basal values within 24 hours after injection. CONCLUSIONS: In humans, Nal-Lys has similar potency and duration of action to other antagonists and produces fewer local side effects. However, the utility of Nal-Lys is limited by formulation difficulties; current efforts are directed at improving the formulation in order to explore the potential clinical uses of this peptide. PMID- 8405526 TI - Overtraining affects male reproductive status. AB - OBJECTIVE: To substantiate the hypothesis that strenuous exercise disrupts the hypothalamic-pituitary-gonadal axis in men. DESIGN: Longitudinal study. SETTING: Normal human volunteers in an academic research environment. PATIENTS: Five endurance-trained men (maximum oxygen consumption 65.4 +/- 3.6 mL/kg per minute [means +/- SEM]) with normal spermatogenic and hormonal profiles. INTERVENTIONS: Semen and blood samples were collected bimonthly before, immediately after, and 3 months after overtraining, which was defined as twice the previous average weekly training volume with unchanged intensity. MAIN OUTCOME MEASURE: Testosterone, cortisol, and sperm concentration. RESULTS: Basal T levels decreased to 5.37 +/- 67 ng/mL from 8.68 +/- 93 ng/mL (conversion factor to SI unit, 3.47) immediately after overtraining and basal cortisol levels increased to 215.3 +/- 31 ng/mL from 145.7 +/- 27 ng/mL (conversion factor to SI unit, 2.76). This inverse relationship was highly correlated (r = -0.92). Both cortisol and T levels returned to pretraining values 3 months after resumption of previous training volume. Sperm count (91 +/- 23.3 x 10(6)) decreased significantly by 43% immediately after overtraining (52 +/- 6.8 x 10(6)) and by 52% 3 months after overtraining (44.5 +/- 20 x 10(6)). However, all values remained within normal range and would not be expected to affect fertility. CONCLUSIONS: Our results indicate that overtraining reduces T levels, which is highly correlated with an increase in levels of cortisol and possibly a subsequent decrease in sperm concentration 74 days later. PMID- 8405527 TI - Ultrasonographic diagnosis of varicoceles. AB - OBJECTIVE: To assess the ability of color duplex scrotal ultrasonography to detect subclinical varicoceles and confirm the diagnosis of clinical varicoceles. DESIGN: Physical examination, color duplex scrotal ultrasonography and internal spermatic venography was performed on 64 testicular units in 33 men. SETTING: Male fertility center. PATIENTS: Two hundred sixty-two consecutive men being evaluated for male factor infertility of whom 33 agreed to undergo venography. MAIN OUTCOME MEASURES: Ultrasonographic measurement of scrotal vein diameter of patients in the supine and upright position, before and during valsalva maneuver, and scrotal vein blood flow reversal with valsalva maneuver was compared with the findings of varicocele by physical examination and venography. RESULTS: The best predictor of a varicocele was internal spermatic vein diameter, and the best overall performance of ultrasonography was achieved with the patient at rest in the supine position. The best cutoff point for venous diameter for a clinical varicocele was 3.6 mm and 2.7 mm for a subclinical varicocele, but the overall accuracy was only 63%. CONCLUSIONS: Confirmatory studies are needed to support the ultrasonographic diagnosis of varicoceles before considering surgical repair. PMID- 8405528 TI - Folinic acid in the treatment of human male infertility. AB - OBJECTIVE: To investigate the clinical and histologic effects of folinic acid in the treatment of round cell idiopathic syndrome. DESIGN: Sixty-five males of infertile couples have been treated with folinic acid alone (15 mg one time per day) for a 3-month period. Various sperm parameters (spermatozoa number and motility, round cell number) have been evaluated before and after the therapy. Moreover, to investigate the histologic patterns of the testis induced by the administration of folinic acid, an experimental animal model has been designed on rats previously treated with chemotherapy drugs. RESULTS: All the investigated human seminal parameters presented significant variations after the treatment. An improvement in spermatozoa number and motility and a decrease in round cell number have constantly been noticed; the significance of the changes was particularly impressive in those men whose partners became pregnant later on. A consistent improvement in the histologic structure of the rat tubular epithelium was also detected. CONCLUSION: Folinic acid appears to be a valuable approach for the treatment of round cell idiopathic syndrome. PMID- 8405529 TI - Quality assurance for sperm concentration using latex beads. AB - OBJECTIVE: To provide a simple, universally applicable method of quality assurance for sperm counting, thereby reducing intercounting chamber variation. DESIGN: By using a known concentration of latex beads, the sperm:bead ratio can be used to calculate the actual sperm count. MAIN OUTCOME MEASURES: The mean sperm and bead counts were determined in both a Spot-lite hemocytometer (Baxter Diagnostics, McGaw Park, IL) and a Makler chamber (Polymedco Inc., Yorktown, NY) from 21 different ejaculates mixed with a known concentration of beads. The hemocytometer chamber was used as the standard counting chamber because it consistently yielded a low variation in sperm count. The adjusted sperm concentration of the Makler chamber was calculated using the following formula [hemocytometer beads]/[Makler beads] x [Makler sperm]. RESULTS: Observed mean +/- SD sperm counts were significantly different between the hemocytometer chamber (110.6 +/- 66.2 x 10(6)/mL) and Makler chamber (173.3 +/- 103.5 x 10(6)/mL). However, calculated Makler chamber sperm counts (118.1 +/- 76.1 x 10(6)/mL) was not statistically different from observed hemocytometer sperm counts. CONCLUSION: This novel approach to sperm counting using a known concentration of latex beads as a reference material can be used to reduce variation in sperm counting between observers, counting chambers, and possibly computerized sperm analyzers. PMID- 8405530 TI - Improvement of post-thaw sperm motility in poor quality human semen. AB - OBJECTIVE: To find alternative cryopreservation methods to improve the post-thaw fertilizing capacity of poor quality human sperm. DESIGN: Controlled clinical study. SETTING: Fertility clinic of a teaching hospital. PATIENTS: Men with poor quality semen samples, i.e., asthenozoospermia (< 40% motile sperm) and/or oligozoospermia (< 20 x 10(6) sperm/mL). Fertile sperm donors were used for comparison. INTERVENTIONS: Semen samples were divided into four aliquots and slowly diluted 1:1 with: [1] n-tris (hydroxymethyl) methyl-2-amino ethane sulfonic acid (TES) and tris (hydroxymethyl) aminomethane (Tris)-citric acid-egg yolk buffer with 12% glycerol (TEST), [2] TEST+CryoSeeds (Cell Systems, Ltd., Cambridge, UK), [3] TEST + 10 mM dithiothreitol (DTT), or [4] TEST+CryoSeeds + 10 mM DTT. Cryovials were frozen using slow staged cooling and static vapor freeze and stored at -196 degrees C. MAIN OUTCOME MEASURE: The frozen aliquots were randomly thawed and, after 15 minutes at 37 degrees C, motion analysis was performed. RESULTS: The percent motility after freeze-thaw in TEST was significantly decreased to 42 +/- 5% of prefreeze motility (P < 0.001). Addition of CryoSeeds with holding at -5 degrees C for 10 minutes resulted in 47 +/- 6% of prefreeze motility, which was not different than TEST alone. Addition of DTT to TEST significantly improved post-thaw motility over TEST alone to 71 +/- 7% of initial motility (P < 0.01). The combination of CryoSeeds and DTT further improved post-thaw motility to 80 +/- 10% of initial motility, which was not different than the neat semen. CONCLUSION: The present results suggest that DTT, a reducing agent that prevents oxidation of sulfhydryl groups, protects poor quality spermatozoa from excessive cryodamage. Thus, DTT along with seeding may be a useful addition when long-term storage of poor quality semen is crucial for maintaining reproductive potential. PMID- 8405531 TI - Platelet activating factor and pentoxifylline as human sperm cryoprotectants. AB - OBJECTIVE: To explore whether the addition of platelet-activating factor (PAF) or pentoxifylline before cryopreservation improves the recovery of motile viable sperm and what role cyclic adenosine monophosphate (cAMP) plays in this recovery. DESIGN: Washed sperm was cryopreserved in the absence of and in the presence of PAF and pentoxifylline. After 2 weeks these samples were quick-thawed and evaluated before and after washing for sperm motility and other motion characteristics. Sperm viability and cAMP concentration were determined to compare the effects of these cryoprotectants. RESULTS: When sperm samples were cryopreserved in the presence of PAF or pentoxifylline, an improvement in the recovery of motile sperm in unwashed and washed post-thaw samples was observed. There were 38% more motile sperm recovered with PAF and 15% more with pentoxifylline when compared with untreated samples. In comparison with the unwashed samples, sperm motility in post-thaw samples was lowered by the washing procedure. When PAF was used as a cryoprotectant, a significant improvement in the linearity and straight line velocity of the post-thaw sperm was observed. When pentoxifylline was used as a cryoprotectant, lateral head displacement was significantly improved in the post-thaw samples than in the control group. Both PAF-and pentoxifylline-treated samples contained a greater number of viable sperm than the control. The cAMP concentrations in post-thaw samples were 12-fold higher in pentoxifylline-treated samples and 4-fold higher in PAF-treated samples when compared with the untreated control. A 4-fold decrease in cAMP concentration was observed in post-thaw control samples compared with fresh-washed sperm. CONCLUSIONS: The results of this study suggest that both PAF and pentoxifylline are useful cryoprotectants for the increased recovery of motile, viable sperm. Although increased recovery of motile sperm in pentoxifylline-treated samples is related to higher cAMP levels, the cryoprotective effect of PAF does not appear to be due to increased cAMP. PMID- 8405532 TI - A successful assisted reproductive technology satellite program. AB - A satellite program was implemented to provide IVF to interested infertile couples living in a five-state area. Thirty-six gynecologists were established as satellite sites after appropriate screening and training. No attempt was made to interchange reference serum specimens between laboratories. Patients were seen at Mayo Clinic for an initial consultation and did not return until the ovulatory dose of hCG was administered. There was no difference in cancellation, clinical pregnancy, and delivery rates between satellite and central unit monitored patients. Satellite monitoring decreases patient inconvenience and time away from home and the workplace without compromising cycle outcome. PMID- 8405533 TI - Clinical outcomes of pregnancies achieved by microinsemination by sperm transfer. AB - Micromanipulative techniques such as microinsemination by sperm transfer are evolving as treatment for couples suffering from some forms of male factor infertility where previously donor insemination was the only option. Being a relatively new technology, the numbers of microinsemination by sperm transfer pregnancies that have progressed to term are still small. However, at this time, it would seem that outcome in terms of antenatal complications, gestation length, and birth weight is no different from that seen with conventional IVF. Continuing review of the rate of congenital malformation in microinsemination by sperm transfer progeny seems warranted. PMID- 8405534 TI - Recombinant human follicle-stimulating hormone treatment leads to normal follicular growth, estradiol secretion, and pregnancy in a World Health Organization group II anovulatory woman. AB - A first case of successful induction of ovulation with recombinant human FSH administered subcutaneously to a WHO group II anovulatory patient is reported. Using a chronic low-dose protocol, recombinant human FSH, a preparation with no LH activity, led to a clear and even supraphysiological increase of inhibin and E2 and the development of four follicles among which one was dominant. The treatment cycle resulted in an ongoing pregnancy. PMID- 8405535 TI - Results of a survey of carrier women for the galactosemia gene. AB - A survey of 108 heterozygote women for the classic galactosemia gene, GALT, did not reveal that the carrier state was associated with premature ovarian failure or ovarian cancer. This survey did not support previous epidemiologic studies suggesting an increased risk for ovarian dysfunction in women with deficiency of the GALT enzyme. PMID- 8405536 TI - Brachial plexus neuropathies after advanced laparoscopic surgery. AB - A retrospective review of 3,200 advanced laparoscopic procedures demonstrated five brachial plexus injuries during a 5-month period in 1986 (0.16% incidence rate). Brachial plexus injury can occur during laparoscopic surgery using steep Trendelenburg's position with shoulder braces and the patient's arm extended at 90 degrees. Position modification can reduce the risk for upper extremity neuropathies. PMID- 8405537 TI - Fertility and the use of assisted reproductive techniques in the adult male exstrophy/epispadias patient. AB - These three cases exemplify, in increasing order of complexity, how the common sense application of already successful techniques might be applied to augment the chance of pregnancy in this group of patients. We believe this to be the first report specifically outlining measures that may assist this group in their goal of pregnancy achievement. To our knowledge, vasal sperm aspiration has never been used for this indication. PMID- 8405538 TI - Kallmann's syndrome: pregnancy achieved through gamete intrafallopian transfer. AB - This case report describes the use of GIFT to achieve pregnancy for a man with Kallmann's syndrome who obtained only marginal sperm counts with both the pulsatile GnRH infusion pump and gonadotropin injections. Failure of this man to achieve a pregnancy with hormonal therapy alone and in combination with IUI suggests that assisted reproductive technologies should be considered in male patients with Kallmann's syndrome when suboptimal sperm concentrations are achieved despite exogenous hormonal stimulation. PMID- 8405539 TI - [Effect of a new pharmaceutical form of fenigidin on the coronary circulation and left ventricular contractility in dogs with acute myocardial infarction]. AB - The experiments on dogs with acute myocardial infarction have revealed that fenigidine, a new pharmaceutical form of calcium antagonist, does not provoke a deterioration of the left ventricular function, but normalizes it. The coronary blood flow greatly increases under the influence of remedy (more than 3 hours). PMID- 8405540 TI - [Histoautoradiographic and morphometric analysis of myocardial tissue in optimal regimen of hyperbaric oxygenation]. AB - The experiment on 90 rabbits and 30 rats has been conducted to compare the effect of different regimes of hyperbaric oxygenation (HBO) (pressure of oxygen: 3 ata, 2 ata, oxygen-air mixture: 3 ata and 1 ata, respectively) on the morphofunctional state of intact and ischemic myocardium with experimental infarction. The histoautoradiographic method using 3H-thymidine and 3H-uridine, the precursors of nucleic acids' synthesis, proved a good criterion of viability and activity of the regenerative processes in cardiomyocytes of perinfarction and necrosis distant zones of the heart. The best effect has been obtained when using HBO regime with oxygen pressure 2 ata, the highest damaging effect--oxygen (3 ata) air (1 ata) mixture. The use of HBO has prevented the development of necrosis foci and dystrophic changes in the auricles. PMID- 8405541 TI - [Activation of free-radical processes as a factor of ionizing radiation-induced changes in contractile activity of a vascular wall]. AB - The influence of ionizing irradiation (1, 2 and 4 Gy 137Cs) on both the activity of free-radical processes in plasma, formed elements and aorta wall as well as on the character of contractile vascular reactions of isolated rings of thoracic aorta and carotid artery in rabbits has been studied. The experiments were carried out on the 7th day after the whole-body irradiation. The results indicate that simultaneously with the weakening of antioxidant mechanisms both endothelium dependent and endothelium-independent vascular wall relaxation slightly decreases after 1 Gy exposure. Noradrenaline and KCI-induced contraction is shown to increase. However, these changes are not statistically significant. Irradiation in dose of 2 and 4 Gy considerably decreases endothelium-dependent relaxation. Nitroglycerin-induced relaxation greatly diminishes, KCI- and noradrenaline induced constriction considerably increases in these conditions. The level of activation of free-radical processes considerably increases too. Thus, already on the 7th day after irradiation significant changes in reactivity of vascular wall are developed. Radiation injures both endothelium and vascular smooth muscle cells. The free-radical processes seem to be the main cause of radiation vascular damage, so there is a pronounced correlation between the changes of vascular contractile properties and the degree of activation of these processes. PMID- 8405542 TI - [Effect of calcium antagonists on the hemodynamic and pancreatic secretory parameters in diabetes mellitus]. AB - Data are presented concerning the basal levels of parathormone and calcitonin and the effect of dihydropyridine derivatives on the characteristics of haemodynamics as well as pancreatic secretory activity in patients with diabetes mellitus. It is shown that in diabetic patients with hypertension a single administration of the drugs induces vasodilatation which causes lowering of systolic and diastolic blood pressure. In patients with diabetes mellitus in combination with hypertension, heart coronary disease and obvious peripheral angiopathy (similar procedure) no peripheral vasodilatation was observed. No negative effects were observed on the compensatory processes (and obviously on pancreatic secretory activity in patients with both insulin-dependent and insulin-independent diabetes mellitus even during long-lasting administration of drugs. A conclusion is made that calcium channel antagonists should be recommended to patients with diabetes mellitus taking into account the state of their peripheral vessels. PMID- 8405543 TI - [Effect of lateral septal nucleus damage on the photoperiodic changes in the reproductive system of male rats]. AB - The experiments on the juvenile male Wistar rats (4-5 weeks old) were carried out to show the effects of damage of the lateral septal nucleus (LSN) on the gonads of rats caged under different photoperiodic regimes: natural change of light and dark phases of the day, constant light and darkness for 24 hours. A short (week) exposition of rats to the permanent light or darkness stimulates or inhibits gonads, respectively. Lesion of LSN in rats caged under permanent light or darkness inhibits the morpho-functional state of the reproductive system and hormone-producing function of testes. The changes are much more pronounced in rats caged under permanent darkness. The possible mechanisms of LSN influence on the photoperiodic changes of the reproductive system of male rats are under discussion. PMID- 8405544 TI - [Aging of the respiratory system in various groups of the population]. AB - Two groups of practically healthy elderly and old people were investigated. The first group consisted of 265 aborigens of three Abkhazian villages examined under field conditions, and the second group was composed of 227 inhabitants of Ukraine, the patients of clinic at Institute of Gerontology in Kiev. It was established that changes of the lung ventilation parameters typical of ageing were less pronounced in the Abkhazian inhabitants than in the Ukrainian ones. Ageing dynamics and values of the studied parameters were the same in Abkhazians: relatives of the long-living people and Abkhazians without hereditary predisposition to longevity. This fact permits a conclusion that the peculiarities of the respiratory system ageing in the Abkhazian population are mainly due to the environmental factors. PMID- 8405545 TI - [Role of thymocytes in the correlation between the thymus and hypophysis]. AB - The influence of the thymus on the growth hormone (GH) level in the mouse hypophysis has been studied. It has been found, that syngeneic transfer of thymocytes of young mice into thymectomized mice causes on increase of the growth hormone level in the hypophysis. Administration of hydrocortisone simultaneously with transfer of thymocytes accelerated the GH level increase. The administration of only hydrocortisone into thymectomized mice does not lead to changes in the GH level. The increased GH level was observed when thymocytes were replaced by some structures shedding from thymocytes after their stimulation with short peptide thymic hormone. It is supposed that thymocytes can stimulate GH accumulation in the hypophysis. PMID- 8405546 TI - [Nonspecific immunosuppressive spleen factor, induced in mice by bacterial immunization]. AB - Frozen spleen of mice immunized with vaccine from S. sonnei has been homogenized in the isotonic saline solution and centrifuged. That supernatant has been shown to inhibit antibody immune response to thymus-dependent antigens (sheep and rabbit erythrocytes) and hypersensitivity of the delayed type to splenocytes of guinea pig. The supernatant did not inhibit the antibody formation to thymus independent antigen (bacterial lipopolysaccharide). Immunosuppressive activity of the supernatant disappeared after treatment with anti-immunoglobulin (but not anti-O-antigen) immunosorbent. The spleen of immunized mice is concluded to contain a factor which inhibits T-cell link of the immune system and is of the immunoglobulin nature. PMID- 8405547 TI - [Characteristics of functional and morphologic liver changes in toxic hepatitis with differing resistance to hypoxia]. AB - The functional and structural changes in the liver of white rats both low- and high-resistant to hypoxia, have been studied using the biochemical and morphological methods. It has been found that intensity and degree of the liver injury in experimental toxic hepatitis depend on the systemic resistance to hypoxia. The functional and morphological changes in the injured liver have been found to be less pronounced in the high-resistant rats as compared with those in the low-resistant animals. PMID- 8405548 TI - [Ultrastructural changes of the lungs and kidneys in endotoxin shock]. AB - Ultrastructural changes in the lungs and kidneys in the dynamics of endotoxin shock have been studied in experiments on rats, rabbits and dogs. It has been shown that in the initial period of shock in the epithelial cells of alveoli and convoluted tubules dystrophic processes take place and simultaneously the vascular permeability increases. In the intermediate period of the endotoxin shock the mesenchymal dystrophy is additionally registered in the lungs, while necrosis of epithelium of some tubules--in the kidneys. The interstitial fibrosis and mitosis of pneumocytes of type II, substituting the respiratory epithelium are revealed in the lungs at the stage of late endotoxemia. The adaptive changes, arising as a result of improving renal hemodynamics are observed in the kidneys. PMID- 8405549 TI - [ACTH-producing function of the adenohypophysis and adrenal weight in the offspring of female rats with chronic liver injury]. AB - ACTH-producing function of adenohypophysis in posterity of female rats with chronic liver injury in different periods of early postnatal ontogenesis has been examined as well as the functional state and reserve potentialities of adenohypophysis in case of immobilized stress. The obtained findings indicate the decreased reserve potentialities of the hypophyso-adrenal system in the process of adaptation to stress loads. PMID- 8405550 TI - [Effect of regulatory renal polypeptides on hemocoagulation and lipid peroxidation in fluoride intoxication]. AB - Polypeptide cytomedine, isolated from renal tissues has been studied for its regulatory effect on hemocoagulation and lipid peroxidation with fluoric intoxication (FI). FI was caused by inoculation of laboratory animals (guinea pigs) with sodium fluoride (100 mg/kg) for 14 days. Following it polypeptide (0.1 mg/kg) was introduced intramuscularly for 7 days. The development of FI was expressed by hypercoagulation delay of fibrinolysis with para-coagulation products appearing in blood decrease of antiaggregation activity of the renal tissue. These phenomena were estimated as manifestations of the first phase of disseminated intravascular blood coagulability (DIC-syndrome). The above reactions proceeded simultaneously with lipid peroxidation activation decrease of the antioxidant protection. The necrotic-dystrophic processes developed in renal and hepatic parenchyma. The renal peptide-cytomedine induced the normalization of lipid peroxidation in blood and renal tissues and fibrinolysis, the decrease in the concentration of para-coagulation products. The pathological changes decreased both in the renal and hepatic tissues. It is possibly, a result of the normalization of secretion and reabsorption in the kidneys. Thus, cytomedine of the kidney exerts a pronounced regulatory and protective effect in the case of acute renal pathology. These results correspond to the conception of the peptidergic organism regulation. PMID- 8405551 TI - [Effect of limbic structure and striatum damage caused by kainic acid on seizure reactions in animals after intracranial injury]. AB - It has been established that hippocampus, enthorhinal cortex, amygdala and substantia nigra (pars reticulata) lesions before head injury lead to a decrease of kainic acid-induced behavioral and electrographic seizure expressions. It can be concluded that after head injury the activation of limbic structures excitability due to excitation of "inputs" to these formations takes place. The obtained data indicate the significant role of nucleus caudatus in activation of posttraumatic brain excitatory mechanisms. PMID- 8405552 TI - [Prostaglandins content in the blood of newborns with various types of hypoxia]. AB - The content of prostaglandins (Pg) E, F2, 6-keto-F1 alpha, thromboxane B2 has been examined in funic blood serum and peripheral blood of 172 newborns. 38 neonates endured chronic hypoxia in the antenatal period, 42 had acute hypoxia during labour and in 39 acute hypoxia during labour developed against a background of chronic intrauterine hypoxia. The control group consisted of 53 infants that were not subjected to hypoxia during antenatal and intranatal periods. In control group in the dynamics of early neonatal period a decrease of PgE, PgE2 alpha concentration and an increase of 6-keto PgF1 alpha, TxB2 were observed. The PgE level was determined to be higher in infants with chronic hypoxia and acute hypoxia than that in the control group. In acute hypoxia the PgE concentration did not differ from that in the control group. PgF2 content was decreased in infants with chronic hypoxia and increased with acute one. The PgF alpha 1 level was noticed to decrease in chronic hypoxia and acute hypoxia against the chronic background. Thromboxane B2 concentration was increased in all the types of hypoxia. The 6 keto PgF1 alpha/TxB2 ratio decreasing in infants with all the types of hypoxia underwent the most pronounced changes. The largest deviations were marked in infants with associative hypoxia. The revealed dynamics of prostaglandin changes in the postnatal period in the control group is similar to the dynamics that reflects the course of stress-reaction under the influence of other stimuli. It permits affirming that there are common mechanisms of the prostaglandin system response to stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405553 TI - [Correlation of chemiluminescent parameters of blood in non-irradiated animals with their survival after irradiation]. AB - Blood chemiluminescence is a reflection of the free radical oxidation process in every organism. The changes of its intensity, as a consequence of changes in the rate of this process, are connected, in its turn, with the changes, in the functional state of the organism. It is supposed that the differences in the radiosensitivity depend on the conditions in which free radical oxidation processes take place. Therefore, the indices of blood chemiluminescence of intact rats have been studied. The aim of the investigation is to find the correlations between these indices and the survival of animals irradiated with gamma-quantas in doses of 3.5, 7.0 and 9.0 Gr. It is established that the coefficients of the reaction rate (k1, k2 and k3) most reliably correlate with survival, whereas no correlation is observed between the other indices of chemiluminescence and survival. PMID- 8405554 TI - [Content of selected prostanoids in the rabbit myocardium as effected by izadrin]. AB - The content of stable metabolites of prostacyclin and thromboxane A2 in the rabbit myocardium in normal conditions and after single izadrin injection (comparison of survived and dead animals) has been comparatively analyzed. It has been shown that in all the cases izadrin injection induces the synthesis of these prostaglandins. But the cases with lethal outcome are characterized by predominance of the thromboxane-synthetase activity. The revealed peculiarities of prostaglandins synthesis in myocardium need experimental check on the isolated heart. PMID- 8405555 TI - [Changes of energy metabolism and concentration of cyclic nucleotides in the kidneys in acute fluoride intoxication and hyperbaric oxygenation]. AB - The energy metabolism and indices of the adenylate-cyclase system in kidneys of rats with acute fluoric intoxication have been studied. It has been determined that the use of hyperbaric oxygenation prevents deep disturbances of energy metabolism in the renal tissue and restricts the increase of adenosine monophosphate concentration. Moreover the survival rate of animals has increased by 30% for the first 24 hours. PMID- 8405556 TI - [Comparison of the effectiveness of oil and water solutions of alpha-tocopherol as radiation protection agent]. AB - The change in alpha-tocopherol content (as an index of the antioxidative organism system mobilization) in the brain, liver, muscles and heart of rats prior to and after total gamma-irradiation with lethal dose has been studied. Prior to the irradiation alpha-tocopheryl-acetate (oil solution), alpha-tocopheryl-phosphate dipotassium salt (water solution), gammaphos (prepared WR 2721, water solution) were introduced into rats. It is shown that water-soluble form of vitamin E is more effective than gammaphos and much more efficient than oil form of alpha tocopherol. PMID- 8405557 TI - [Effectiveness of various pharmaceutical forms of alfa-tocopherol]. AB - The content of alpha-tocopherol in brain, liver, heart and muscle of Wistar rats has been studied. The water-soluble form of alpha-tocopherol: alpha-tocopheryl phosphate dipotassium is more effective than the oil-soluble form: alpha tocopheryl-acetate. PMID- 8405558 TI - [Mechanisms of therapeutic and preventive effects of adaptation to hypoxia in arterial hypertension]. AB - Data on the effect of adaptation to hypoxia on the cardio-vascular system responses and arterial pressure regulation have been considered. The role of sympathoadrenal system rearrangement in regulation of cardiac output and vascular tension has been analyzed. The data on the possible role of water-salt metabolism changes in a decrease of arterial pressure after long-term effect of hypoxia are presented. It is concluded that hypoxic training is a promising trend of prevention and treatment of arterial hypertension. PMID- 8405559 TI - A special message to our future colleagues. PMID- 8405560 TI - The legal regulation of nursing practice. PMID- 8405562 TI - The nurse practitioner in college health. PMID- 8405561 TI - Culture shock in nursing school. PMID- 8405563 TI - Advanced clinical practice in the emergency department. PMID- 8405564 TI - The importance of change. PMID- 8405565 TI - Looking beyond the apparent. PMID- 8405566 TI - Mastering the practice of the art of nursing. PMID- 8405567 TI - Distribution of dividing thymocytes in rat thymic cortex during daily cycle. AB - In the present study, frequency of thymocyte divisions was analyzed in the rat thymic cortex, as related to (1) distance from thymic capsule and (2) the daily cycle. Frequency of thymocyte divisions was estimated in successive 10 micron(s) thick layers of the cortex. An evident daily cycle was detected in the thymic cortex with increase in the number of cell divisions around 8 pm. PMID- 8405568 TI - Effect of long-term exposure to lead on testis and epididymis in rats. AB - The relation was studied between the morphology of tests and epididymides on the one hand, and the lead content in these organs on the other. The testes of rats, which for the time of 5 spermatogeneses (9 months) were drinking 1% lead acetate(II), displayed all generations and layers of spermatogenic cells at respective stages of the seminiferous epithelium cycle. The lead content in testes of the animals did not differ significantly from the value of this element in gonads of control rats. The epididymal cells also failed to show morphological changes however, in all epididymal zones there were fewer spermatozoa than in the corresponding zones in control rats. Many spermatozoa revealed abnormal reactions of oxidoreductive enzymes in the midpiece. There were also local defects in tetrazolium salt reduction and segmental or total lack of formazan deposits in the mitochondrial sheath. The lead content in epididymes of these animals was significantly higher than in epididymides of control rats. The obtained results of the studies highlight the possibility for lead to accumulate in epididymis, and for lead compounds to damage spermatozoa in this organ. PMID- 8405569 TI - Immunogenic and nondividing mafosfamide-treated L 1210 cells--comparison of lines resistant and nonresistant to cyclophosphamide. AB - Cyclophosphamide/mafosfamide-resistant L 1210 cell line [L 1210(Cy)R] was established from a sensitive parental line. The L 1210(Cy)R line was resistant to cyclophosphamide at the dose of 100 mg/kg. Cells of L 1210(Cy)R line were more immunogenic for semisyngeneic CD2F1, mice as compared with parental line. They grew slower in immunocompetent mice compared to immunosuppressed mice. It has been shown that L 1210(Cy)R cells treated with mafosfamide at high concentration not only retained immunogenicity but were even more immunogenic than parental L 1210 cells. In conclusion, it was possible to produce immunogenic, nondividing leukemia cells even when cells were resistant to the cytostatic used for cell modification. PMID- 8405570 TI - Reversible inhibition of growth of B16 melanoma cells by decreased amino acid nutrition. AB - The effect of limited amino acid nutrition upon the growth of B16 melanoma cells was investigated. It was observed that the cells in Basal Medium Eagle do not proliferate and are inhibited predominantly in the G1 phase of cell cycle. The cells which were inhibited in growth for 7 or 12 days in Basal Medium Eagle were able to commence proliferation when transferred to Eagle's Minimal Essential Medium containing higher concentration of amino acids. The proliferating cells were found to be more evenly distributed throughout all phases of the cell cycle. The reversible inhibition of growth of B16 melanoma cells was caused by decreased concentrations of amino acids and not by their depletion in the culture medium. The B16 melanoma cells which are known to be capable of highly autonomous growth in the absence of extracellular hormones and growth factors have appeared to be sensitive to nutritional inadequacy. PMID- 8405571 TI - Inactivation and denaturation of some proteins by enzyme system: myeloperoxidase, chloride and hydrogen peroxide. AB - Changes in biological properties of serum albumin, egg white lysozyme, human serum alpha-1 antiproteinase and human leukocyte ribonuclease in effect of interaction with the enzyme system composed of myeloperoxidase from human neutrophilic polymorphonuclear leukocytes, Cl- and H2O2 were investigated. All the studied proteins lost their biological functions and were denaturated, but the amounts of hydrogen peroxide necessary to produce these effects differed remarkably for each individual protein. The alpha-1 antiproteinase ability of binding to trypsin was abolished upon employing 1.2 mols of H2O2 per mol of alpha 1 antiproteinase. The lysozyme enzymatic activity was abolished when 1.4 mols of H2O2 per mol of lysozyme were employed. Albumin decreased its binding to specific antialbumin antibodies and entirely lost the binding properties when 2 mols and about 10 mols of H2O2 per mol of albumin were employed, respectively. On the other hand 18 mols of H2O2 per mol of human leukocyte ribonuclease were necessary to inactivate this enzyme. All the mentioned proteins were protected from losing their biological functions by excess of specific amino acids with affinity to hypochlorite: Alpha-1 antiproteinase by excess of N-acetylmethionine, lysozyme by N-acetylmethionine and N-acetyl glycyltryptophane, albumin by N-acetyl derivatives of methionine, cysteine, tryptophane and lysine, whereas ribonuclease was protected from denaturation by all above mentioned amino acid derivatives. None of the studied proteins was protected from denaturation by N-acetyl tyrosine, or phenylalanine. PMID- 8405572 TI - Cell proliferation in colonic crypts of germ-free and conventional mice- preliminary report. AB - The influence of normal microbial flora on cell proliferation in mouse colonic crypts was assessed by a comparison of conventional and germ-free animals. The following characteristics of the crypts were measured: length, diameter in the middle, number of columns, number of cell per one column, percentage of goblet cells, mitotic and labelling index distribution and average cell cycle time for the proliferative compartment. It has been found that the crypts in conventional mice are longer than in their germ-free counterparts, while their diameter and percentage of goblet cells were similar in both animal groups. The mitotic index was lower in germ-free mice, as well as the labelling index. The average cell cycle time was about 1.7-fold shorter in conventional animals than in germ-free animals. This result was verified using computer simulation model; it turned out that the change in average cell cycle time could be solely responsible for all the observed differences between germ-free and conventional animals. It seems therefore that normal microbial flora stimulates cell proliferation in mouse colon, most probably due to shortening of the average cell cycle. PMID- 8405573 TI - Correlation between the occurrence of Ki-67 antigen and clinical parameters in human breast carcinoma. AB - Cryostat sections of ductal breast carcinomas obtained during surgery from 48 patients were subjected to an indirect immunostaining with an antibody against Ki 67 antigen (Dako). The primary reaction was visualized by the APAAP technique with new fuchsin as chromogen. The percentage of positively stained nuclei was correlated with tumor size, number of metastatic lymph nodes, presence or absence of metastases in distant organs as well as with patient's age. The best correlations were achieved when the data were considered separately within the groups with low, medium and high Ki-67 values. A possible significance of Ki-67 evaluation for the prognosis and therapy is discussed. PMID- 8405574 TI - Structure of ovaries in two collembolans, Allacma fusca and Arrhopalites coecus (Hexapoda, Entognatha). AB - The ovaries of both investigated species are of the polytrophic-meroistic type. They are sac-shaped and not divided into individual ovarioles. Two compartments: the germarium and vitellarium can be discerned within each ovarial sac. The germaria are localized laterally and contain relatively numerous germ cells. In the vitellaria, synchronously developing oocyte-nurse cell complexes occur. In each complex, 3 morphologically, and presumably functionally distinct categories of nurse cells can be distinguished. Follicular cells do not form a continuous follicular epithelium; they are filled with synthetically active organelles and secretory vacuoles. Morphology of the egg envelopes of A. coecus is also described. PMID- 8405575 TI - The role of vacuolar digestive system in preventing cadmium poisoning in Acanthamoeba castellanii. AB - Constituents showing cadmium affinity were found in digestive residues of the vacuolar digestive system in Acanthamoeba. Cadmium was detected by means of X-ray analysis in complexes appearing both in residues included within the digestive vacuoles and in those egested outside the cell. The digestive vacuolar system in Acanthamoeba castellanii is characterized by a particularly high turnover rate (4). Confronted with this fact, results of the present study suggest that the system is able to egest rapidly the ligand-bound cadmium. In this way protects the amoeba against serious toxic effects of the Cd-polluted environment. PMID- 8405576 TI - Fusarium mycotoxins nivalenol and 4-acetyl-nivalenol (fusarenon-X) in mouldy maize harvested from farms in Jos district, Nigeria. AB - Twelve random samples of mouldy maize collected during the 1991 harvest season from farms at different locations in Jos district, Nigeria, were screened primarily for contamination by the Fusarium mycotoxins nivalenol, fusarenon-X and HT-2 toxin. All the three mycotoxins were detected: nivalenol in three samples, (0.8-1.0 mg), fusarenon-X in four samples (3.0-15.0 mg) and HT-2 in only one sample (3.0 mg/kg). Deoxynivalenol (three samples) and T-2 toxin (one sample) were also detected. Deoxynivalenol and nivalenol co-contaminated the same samples, with nivalenol being present at much lower levels than the former. The maize samples were not destined for human consumption. PMID- 8405577 TI - Analysis of Canadian and imported beers for Fusarium mycotoxins by gas chromatography-mass spectrometry. AB - A sensitive method was developed for the determination of deoxynivalenol (DON), nivalenol (NIV), alpha-zearalenol (alpha-ZEL), beta-zearalenol (beta-ZEL) and zearalenone (ZEN) in beer by capillary gas chromatography-mass spectrometry (GC MS) of their heptafluorobutyrate (HFB) derivatives. Recoveries averaging 90-103% were obtained from beers spiked with each mycotoxin in the 5-20 ng/ml concentration range. Limits of detection were 0.1-1.5 ng DON/ml, 0.01-0.3 ng NIV/ml, 2.5-3 ng alpha- and beta-ZEL/ml, and 1.5-2 ng ZEN/ml. Twenty-nine of 50 samples of Canadian and imported beer surveyed were found to contain DON; of these nine contained greater than 5 ng/ml (up to 50 ng/ml). The identity of DON was confirmed by response ratios at m/z 670 and m/z 884 for the HFB derivative and m/z 497 and m/z 512 for the trimethylsilyl (TMS) derivative. NIV was also detected in three beer samples (0.1-0.84 ng/ml) but no alpha-ZEL, beta-ZEL or ZEN was found. PMID- 8405578 TI - Toxicological studies of the false morel (Gyromitra esculenta): embryotoxicity of monomethylhydrazine in the rat. AB - The embryotoxic and teratogenic potential of monomethylhydrazine (MMH), a toxic component of the widely consumed false morel (Gyromitra esculenta), was studied in rat. Groups of pregnant Sprague-Dawley rats received MMH as a constant i.v. infusion via implanted osmotic minipumps (1.2, 3.0, 4.2, 6.0, 9.0 or 13.2 mg MMH/kg bw/day) on days 6-13 of pregnancy, or as a single intragastric bolus (1 mg MMH/kg/bw or 5 mg MMH/kg/bw) on day 6 of pregnancy. Controls received corresponding amounts of saline. The average maternal serum concentrations, measured during the infusion treatment with a sensitive HPLC method, ranged from 0.072 micrograms MMH/ml (lowest dose) to 0.60 microgram MMH/ml (highest dose). The average serum levels measured 45 min after the intragastric application (peak levels) were 0.28 microgram MMH/ml and 1.6 microgram MMH/ml, respectively. Serum concentrations of MMH corresponding to those measured in the lower dose groups in this study were seen in pilot studies after a single mushroom meal in human volunteers. A dose-dependent, statistically significant increase in the number of resorptions was seen in all but the lowest dose group after the infusion of MMH. In addition, except for the two lowest doses, there was a dramatic, dose dependent decrease in the pregnancy rate as compared to controls, with no pregnancies occurring at the two highest dose level groups. The decreased pregnancy rate was probably due to preimplantation loss which was shown to occur after a single intragastric bolus dose of MMH (5 mg/kg bw).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405579 TI - Levels of benzo[a]pyrene and other polycyclic aromatic hydrocarbons in liquid smoke flavour and some smoked foods. AB - In order to investigate the levels of the polycyclic aromatic hydrocarbons, mainly benzo[a]pyrene because of its carcinogenicity, 55 samples of smoke flavour and smoked foods were analysed. The samples tested included 11 samples of liquid smoke flavour and 44 samples of smoked foods like bacon, loin, turkey, sausage, ox rib, etc. from different brands. A liquid chromatographic method was developed using a fluorescence detector. Benzo[a]pyrene was found in 73% of the liquid smoke flavour samples analysed. The levels varied from 0.1 to 336.6 micrograms/kg. Three liquid smoke flavour samples showed levels of benzo[a]pyrene above the maximum level recommended by FAO/WHO (10 micrograms/kg). From the total of 44 smoked food samples analysed, benzo(a)pyrene was detected in 23 samples (52%). The levels varied from 0.1 to 5.9 micrograms/kg. Anthracene and fluoranthene, non-carcinogenic polycyclic aromatic hydrocarbons, were found in almost all the samples analysed. Benzo[ghi]perylene, 3,4-benzofluoranthene and 1,2,3,4-dibenzopyrene were not found in any of the 55 samples analysed. PMID- 8405580 TI - Organochlorine contaminants in Swedish human milk: studies conducted at the National Food Administration 1981-1990. AB - The concentrations of selected organochlorine contaminants in Swedish human milk have been determined in a number of studies conducted at the National Food Administration during the 1980s. It was shown that the time of sampling post partum (subsequent to childbirth) had little effect on the levels of the DDT complex, beta-HCH, HCB and PCBs in breast milk fat. However, the levels of these compounds decreased with increasing parity (number of children born). For this reason it is recommended that primiparae (women nursing their first child) be sampled when comparing levels over time and in milk from mothers from different geographical areas. The levels of organochlorine compounds in the fat of breast milk from primiparae living in different parts of Sweden were similar. Finally, a study on time trends in concentrations of these compounds in milk from mothers living in Uppsala showed that a marked decline in levels has taken place during the 1980s. PMID- 8405581 TI - Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in human milk samples collected across Canada in 1986-87. AB - Monitoring of human milk samples for the presence of chemical contaminants has been carried out in Canada for several decades. In 1986-87 over 400 donations of human milk were collected nationwide and 100 of these, selected by province according to population, were analysed for the PCDDs/PCDFs. The results show the presence of about eleven different analytes all with 2,3,7,8-substitution with the PCDDs at higher and more variable concentrations than the PCDFs. On a 2,3,7,8 TCDD toxic equivalents (TEQ) basis, the mean value for the country is about 15 ng per kg milk fat. This TEQ value is similar to that from many other industrialized nations and about 50% lower than certain European states. Little or no variation was apparent in the milk content among the various Canadian provinces probably reflecting the uniformity of the general food supply across the country. Limited data over a period of five years suggest the average level of these compounds in human milk may be decreasing. PMID- 8405582 TI - Levels of chlorinated hydrocarbon residues in Canadian human breast milk and their relationship to some characteristics of the donors. AB - A total of 412 breast milk samples from women in all provinces of Canada were analysed for polychlorinated biphenyls, eight chlorinated benzenes, 2,3 dichloronaphthalene, Mirex, alpha, beta, gamma and delta hexachlorocyclohexane, alpha and gamma chlordane, oxychlordane, transnonachlor, p,p'-DDT and some analogues, heptachlor epoxide, dieldrin and octachlorostyrene. No delta hexachlorocyclohexane, heptachlor or aldrin were found in any of the samples, while median levels of the 1,2,4- and 1,3,5-trichlorobenzene, 1,2,3,4- and 1,2,3,5-tetrachlorobenzenes, gamma chlordane, o,p'-DDT and octachlorostyrene were all less than the minimum detectable level (MDL). All other compounds were present at median levels ranging from < 0.1 to 7.2 ng/g whole milk, but did not occur in all samples. Di to tetrachlorobenzenes, except 1,2,4-trichlorobenzene, 2,3-dichloronaphthalene and alpha-chlordane were observed in < 50% of the samples. From 1967 to 1986 a steady decline of hexachlorobenzene, gamma hexachlorocyclohexane, DDTs, heptachlor epoxide and dieldrin was observed in Canadian breast milk. In addition, a decrease in some chlorinated benzenes, gamma hexachlorocyclohexane and PCBs was also observed between 1982 and 1986. Levels of oxychlordane and trans-nonachlor remained constant. Canadian breast milk contamination appears to be one of the lowest among the industrialized nations. Regional and provincial differences in residue levels appeared minimal, although more often samples from Quebec and British Columbia had higher levels of some contaminants, such as oxychlordane, than samples from other provinces. No relationship was found between maternal age and residue levels, but some contaminants, such as PCBs, were significantly higher in the milk of mothers who breastfed their first child as compared to multiparous mothers. Breast milk residue levels were not related to fish consumption. PMID- 8405583 TI - Effects of electron beam irradiation on the migration of antioxidants and their degradation products from commercial polypropylene into food simulating liquids. AB - The influence of electron beam irradiation on migrational behaviour of additives present in food packaging material was studied. The migration experiments were carried out on irradiated and non-irradiated polypropylene pouches containing aqueous food simulating liquids (FSL) for 10 days at 40 degrees C. The controls were irradiated and non-irradiated pouches without FSL contact. After the contact period, the polypropylene and the FSL were analysed. A comparison between the results obtained by the two analyses showed the migration of three products of antioxidant degradation from the polypropylene into the FSL, and a partial decomposition of these migrants in the FSL. PMID- 8405584 TI - Plasticizers from printing inks in a selection of food packagings and their migration to food. AB - A survey of plasticizers in printing inks present in a selection of food packaging including confectionery, snacks, crisps, potatoes, chocolate bars and biscuits, has been carried out both in England and Spain. The identification of the polymer used as packaging showed that almost all of them were oriented polypropylene. Printing inks that are always used on the outer surface contain phthalates as major plasticizers, N-ethyl- and N-methyl-toluenesulphonamides and tris(2-ethylhexyl)trimellitate were found in some of the samples studied. Several food samples were also analysed to check the level of plasticizers migrating from printing inks. PMID- 8405585 TI - Sulphonamide and dapsone residues in bovine milk following intramammary infusion. AB - The elimination into bovine milk of sulphonamides (sulphadimidine and sulphamethoxypyridazine) and dapsone following intramammary infusion was studied. Determination of sulphonamides and dapsone in milk was performed by a high performance liquid chromatographic method with UV detection. The limit of quantification was 0.01 microgram/ml. Withdrawal times were established considering the maximum residue limits fixed by the European Community (100 micrograms/kg for sulphonamides and 25 micrograms/kg for dapsone). The diffusion of residues into milk from a quarter infused by the intramammary route to the untreated quarters was also studied. PMID- 8405586 TI - Calculated dietary intakes of nitrate and nitrite by young Finns. AB - Dietary intakes of nitrate and nitrite of 1212 Finns aged 9, 12, 15, 18, 21, and 24 years were calculated using food consumption data obtained by the 48-hour recall method in 1986, in connection with the Study on Cardiovascular Risk in Young Finns. Files on nitrate and nitrite content of foods and water were compiled for this study. The mean daily intakes of nitrate (NO3-) and nitrite (NO2-) from food were 54.0 mg and 1.4 mg, respectively. Vegetables including potatoes contributed 86% of nitrate intake and meat products 69% of the nitrite intake. If two litres of water within the mode concentration class of nitrate and nitrite were used, the intake of nitrate from water would have been between 1.0 and 2.0 mg and the intake of nitrite between 0.0 and 0.2 mg. PMID- 8405587 TI - Ochrobactrum anthropi bacteremia in a child. PMID- 8405588 TI - Radiograph of the month. Ewing's sarcoma. PMID- 8405589 TI - More on patient records. PMID- 8405590 TI - Why stop smoking? PMID- 8405591 TI - National Practitioner Data Bank. PMID- 8405592 TI - Update of school health talks program, 1993. PMID- 8405593 TI - Resting sites of Anopheles stephensi Liston in Calcutta. PMID- 8405594 TI - Trends in malaria morbidity and mortality in Sri Lanka. AB - Trends since 1930 in malaria morbidity and mortality in Sri Lanka were analysed. The Malaria Control Programme, which began in 1945 with DDT spraying, was associated with a 100-fold reduction in morbidity and mortality over the following ten years, and gave way to the Malaria Eradication Programme in 1958. DDT spraying ceased in 1964 and a vivax malaria epidemic in 1968 returned to the island to 1952 morbidity levels, though with little mortality. After the discovery of DDT resistance in 1969, malathion spraying took over in 1973, and USAID-assisted control programme, involving case-detection and treatment, started in 1977. However, morbidity levels comparable to 1952 levels were observed in 1975 and 1986 when falciparum malaria morbidity levels were especially high. Mortality rates since 1960 have however remained lower than at any other previous time. PMID- 8405595 TI - Control of mosquito breeding through Gambusia affinis in rice fields. AB - Studies on mosquito breeding and its control through Gambusia affinis in nursery and paddy fields after transplantation of seedlings were carried out during June to October 1991 in about 10 ha rice field area. Six anopheline species, viz. An. culicifacies, An. annularis, An. subpictus, An. nigerrimus, An. barbirostris and An. aconitus, and four culicine species, viz. Cx. tritaeniorhynchus, Cx. bitaeniorhynchus, Cx. quinquefasciatus, and Aedes sp. could be identified. These were found breeding in rice fields with fluctuations in their percentage composition, exhibiting species succession in different months. G. affinis survived well in submerged rice fields and provided 87.8% mosquito larval control. In rice fields which exhibited intermittent drying up leading to formation of pools, puddles etc., moderate larval control was achieved. However, in nursery rice fields, this method was not applicable. Mosquito larval control through larvivorous fish in rice fields can be achieved but the method has limitations. PMID- 8405596 TI - In vitro activity of fluoroquinolones against chloroquine-sensitive and chloroquine-resistant Plasmodium falciparum. AB - The in vitro activity of three fluoroquinolones--ciprofloxacin, norfloxacin and ofloxacin--was studied on four laboratory-adapted strains (one chloroquine resistant) and one fresh isolate of P. falciparum from Delhi by the schizont maturation inhibition microtest. The IC50 concentrations (mean +/- SD) were found to be as: ciprofloxacin 6.38 +/- 1.34 micrograms/ml, norfloxacin 11.24 +/- 1.27 micrograms/ml, and ofloxacin 22.3 +/- 3.11 micrograms/ml, while the MIC values were 32 micrograms/ml, 64 micrograms/ml and 128 micrograms/ml for the three drugs in the same order. The IC50 and MIC values for chloroquine-resistant strain were not significantly different from those for the chloroquine-sensitive strains. We conclude that there is little interstrain variability in the in vitro susceptibility of P. falciparum to fluoroquinolones, and that there is no cross resistance between them and chloroquine. The reported variability in clinical response of falciparum malaria to fluoroquinolones is not likely to be due to variation in parasite sensitivity. PMID- 8405597 TI - Insecticide susceptibility status of Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti in Panaji, Goa. AB - Adult and larval insecticidal tests conducted in Panaji, Goa, revealed that adults of An. stephensi were resistant to DDT (4.0%), dieldrin (0.4%) and malathion (5.0%). The larvae of An. stephensi were also resistant to DDT (2.5 mg/l) and malathion (3.125 mg/l). Cx. quinquefasciatus adults were resistant to DDT, dieldrin, malathion and fenitrothion (1.0%), and larvae were highly resistant to DDT but showed low resistance to malathion and fenitrothion (0.125 mg/I). Adults of Ae. aegypti were resistant to DDT and dieldrin and the larvae showed resistance to DDT but good susceptibility to fenitrothion and complete susceptibility to malathion. The triple resistance of An. stephensi to DDT, dieldrin and malathion is intriguing as malathion has never been used in the health sector in Goa. A very limited use of malathion was done in the agricultural sector in Chandor area, which is 50 km away from the study area. PMID- 8405598 TI - Laboratory and field evaluation of Spherix, a formulation of Bacillus sphaericus (B-101), to control breeding of Anopheles stephensi and Culex quinquefasciatus. AB - Spherix, a powder formulation of Bacillus sphaericus strain B-101, serotype H5a 5b, was evaluated against larvae of Anopheles stephensi and Culex quinquefasciatus in both the laboratory and field. In laboratory tests the formulation @ 0.1 g/sq m produced 100% mortality against larvae of both mosquito species at room temperature (28-32 degrees C). The larvicidal activity of Spherix against An. stephensi @ 0.5 g/sq m persisted for over 12 weeks under laboratory conditions. Field evaluation of Spherix @ 0.25-2.0 g/sq m produced 95-100% reduction in the larval density of both An. stephensi and Culex quinquefasciatus within 48 h in different habitats, and the larvicidal activity persisted for 2-4 weeks in water habitats. PMID- 8405599 TI - Interspecific associations among anophelines in different breeding habitats of Kheda district Gujarat: Part II--Non-canal area. AB - A total of 25,858 anophelines comprising 15 species emerged from 1104 samples of immatures from 9 breeding habitats surveyed for one year in six villages representing the non-canal area of Kheda district. Species diversity and homogeneity were maximum in seepage drains. An. annularis, An. barbirostris, An. culicifacies, An. stephensi and An. subpictus were ubiquitous. Interspecific associations occurred more commonly in habitats representing the stable ecosystem. Community groups of anophelines in different habitats were identified. PMID- 8405600 TI - Sexual activity among never-married men in northern Thailand. AB - We use data collected in 1991 to investigate sexual activity among never-married men in Thailand, with a focus on age at first intercourse, characteristics of sexual partners, and conditions under which men visit prostitutes. We sampled men from a broad spectrum of northern Thai society, including university undergraduates, soldiers, and semi-skilled/unskilled workers. We found that except for the students, the majority of each subsample is sexually experienced; prostitutes are the most common type of sexual partner for all groups. Alcohol consumption is associated with several measures of sexual activity. Condom use with prostitutes varies among the subsamples. Among men who have both prostitute and nonprostitute partners, the majority of those who do not use condoms with prostitutes also do not use condoms with their nonprostitute partners. We consider the implications of these results for the AIDS epidemic in Thailand. PMID- 8405601 TI - Gender preference and birth spacing in Matlab, Bangladesh. AB - Gender preference, particularly son preference, is believed to sustain high fertility in many Asian countries, but previous research shows unclear effects. We examine and compare gender-preference effects on fertility in two otherwise comparable populations in Bangladesh that differ markedly in their access to and use of contraception. We expect, and find, stronger effects of gender preference in the population that has more access to contraception and higher levels of contraceptive use. Thus gender preference may emerge as a significant barrier to further national family planning efforts in Bangladesh. We find that if a woman has at least one daughter, the risk of a subsequent birth is related negatively to the number of sons. Women with no daughters also experience a higher risk of having a subsequent birth; this finding suggests that there is also some preference for daughters. Son preference is strong in both the early and later stages of family formation, but women also want to have at least one daughter after having several sons. PMID- 8405602 TI - Nutrition, lactation, and birth spacing in Filipino women. AB - The Cebu Longitudinal Health and Nutrition Survey is used to examine the roles of women's nutrition and infant feeding in determining time from birth to menses and time from menses to conception. The analysis sample includes 2,648 Filipino women followed for 24 months postpartum. Recently devised statistical estimation techniques to control for unobserved heterogeneity and endogeneity are employed in estimating a two-state hazard model. Low body mass index and lower dietary fat intake are associated with increased duration of postpartum amenorrhea. Contraceptive use, high dietary fat consumption, higher parity, and absence of spouse predict a longer waiting time to conception once menses have returned. Simulation of the hazard model is used to examine the effects of the key nutrition and lactation factors. PMID- 8405603 TI - The economic costs of martial disruption for young women over the past two decades. AB - This paper examines the economic costs of separation and divorce for young women in the United States from the late 1960s through the late 1980s. Broadened opportunities for women outside marriage may have alleviated some of the severe economic costs of marital disruption for women. This paper contrasts the experiences of two cohorts of young women: those who married and separated or divorced in the late 1960s through the mid-1970s and those who experienced these events in the 1980s. Based on panel data from the National Longitudinal Surveys of Youth 1979-1988. Young Women 1968-1978, and Young Men 1966-1978, the results show stability in the costs of disruption. A multivariate analysis shows that young women in the more recent cohort have more labor force experience before disruption than those in the earlier cohort, but prior work history does not protect women from the severe costs of marital disruption. PMID- 8405604 TI - Catholicism and marriage in the United States. AB - This study examines the effects of a Catholic background on age at first marriage, the odds of never marrying, and the odds of ever divorcing. Estimates using Catholic upbringing are compared with estimates using Catholic at the time of the survey. A case is made that if the latter measure of Catholicism is used, serious selection bias problems occur in some cases because this measure excludes defectors and includes converts. Further, it is shown that a Catholic upbringing generally has no effect on men's age at first marriage and has a positive effect on the age when women marry. It is also shown that older Baptist men are substantially more likely than Catholic men to experience a divorce. Older Catholic women are somewhat less likely to experience a divorce than non-Baptist Protestant women. There is no Catholic effect on the odds that younger men and women will divorce. PMID- 8405605 TI - Religion as a determinant of marital stability. AB - Using data from the 1987-1988 National Survey of Families and Households, this paper studies the role of the religious composition of unions as a determinant of marital stability. With the exceptions of Mormons and individuals with no religious identification, stability is found to be remarkably similar across the various types of homogamous unions. Consistent with the notion that religion is a complementary marital trait, interfaith unions have generally higher rates of dissolution than intrafaith unions. The destabilizing effect of out-marriage varies inversely with the similarity in beliefs and practices of the two religions as well as with the mutual tolerance embodied in their respective doctrines. The results also suggest that religious compatibility between spouses at the time of marriage has a large influence on marital stability, rivaling in magnitude that of age at marriage and, at least for Protestants and Catholics, dominating any adverse effects of differences in religious background. PMID- 8405606 TI - The dynamics of smallpox epidemics in Britain, 1550-1800. AB - Time-series analysis, a valuable tool in studying population dynamics, has been used to determine the periodicity of smallpox epidemics during the seventeenth and eighteenth centuries in two contrasting representative situations: 1) London, a large city where smallpox was endemic, and 2) Penrith, a small rural town. The interepidemic period was found to be two years in London and five years in Penrith. Equations governing the dynamics of epidemics predict 1) a two-year periodicity and 2) that oscillatory epidemics die out quickly. It is suggested that epidemics were maintained by a periodic variation in susceptibility linked either to a five-year cycle of malnutrition or to an annual cycle. Computer modeling shows how the very different patterns of epidemics are related to population size and to the magnitude of the oscillation in susceptibility. PMID- 8405607 TI - Measuring the effect of changing legislation on the frequency of divorce: The Netherlands, 1830-1990. AB - This article discusses different procedures for measuring the effects of judicial changes on divorce rates. It presents an alternative model and applies it to a historical time series for the Netherlands. In this model, intervention variables were added to a statistical time-series (ARIMA) model. The conclusion of our analysis was that the effects of three judicial changes were only temporary. PMID- 8405608 TI - The determinants of early fertility decline in Texas. AB - This study examines the determinants of fertility control in a frontier population made up largely of German-Americans during the years from 1850 to 1910. The analysis employs a complex register of population constructed from census enumerations, civil and ecclesiastical vital registration, and tax assessment rolls. The article begins with a series of bivariate analyses with cohort of mother's birth, religion, ethnicity, and husband's occupation determining marital fertility. The second half of the paper presents a multivariate model of the determinants of fertility using these and other demographic characteristics as independent variables. The conclusions emphasize the importance of the overall trend toward fertility decline in the United States, as well as the role of religion and of occupational differences, in determining changes in fertility behavior in the population of Gillespie County, Texas. PMID- 8405609 TI - Using survey data to assess neonatal tetanus mortality levels and trends in developing countries. AB - Demographic and health surveys are a useful source of information on the levels and trends of neonatal mortality in developing countries. Such surveys provide data on mortality occurring at 4-14 days of life, which is a sensitive indicator of neonatal tetanus mortality. We analyze birth history data from 37 national surveys in developing countries to assess the quality of neonatal mortality data and to estimate levels and trends in mortality occurring at 4-14 days. It is shown that mortality at 4-14 days has declined considerably during the last decade in most developing countries, concomitant with development and expansion of programs to reduce neonatal tetanus. These declines show that reductions in neonatal tetanus mortality probably have been an important contributor to the decline of neonatal and infant mortality during the 1980s. PMID- 8405610 TI - Siblings' neonatal mortality risks and birth spacing in Bangladesh. AB - This paper studies the familial association of neonatal mortality in Matlab, Bangladesh and its relationship to birth-spacing effects on mortality. Findings show that familial association is strongest for siblings of adjacent birth orders. Moreover, birth-spacing effects on neonatal mortality are stronger when the preceding child has survived the neonatal period than when it has died. Transitional (Markov), random-effects, and marginal models for correlated data are introduced, and are contrasted in interpretation and technique. Familial association of neonatal mortality can be approximately well by a first-order Markov model using generalized estimating equations (GEE) to allow for higher order correlation. PMID- 8405611 TI - Race, intervening variables, and two components of low birth weight. AB - The low birth weight (LBW) gap between blacks and whites has remained largely unexplained in past research. Most previous research on the topic has focused on LBW as a single entity, and without using a causal framework for analysis. The present study examines the determinants of race differences in the two main components of LBW--preterm birth (PRETERM) and intrauterine growth retardation (IUGR)-within a partially causal framework that includes social and proximate explanatory variables. The data come from the 1988 National Survey of Family Growth. This study asks, first, through what intervening variables do race and other exogenous sociodemographic variables operate to raise the risk of these adverse pregnancy outcomes? Second, what are the determinants of the two components of LBW when all explanatory variables are included in the model? The findings reveal differences in how race (and other exogenous variables) act through downstream variables to affect PRETERM and IUGR, as well as differences in the net determinants of these pregnancy outcomes. The models are better able to account statistically for race differences in IUGR (which is explained by intervening sociodemographic, attitude, and behavior variables) than in PRETERM (which is explained partly by intervening health variables). PMID- 8405612 TI - Radiovisiography: an update. AB - Comparatively little has been published regarding the recently developed (and rapidly developing) digital imaging dental radiographic system known as radiovisiography (RVG). This paper reviews the existing literature, describes various modifications and updates which have been made and discusses the clinical implications of RVG. PMID- 8405613 TI - PTFE tape: a versatile material in restorative dentistry. AB - PTFE is widely used in industry and commercial products, but is not widely used in dentistry. This paper describes some clinical applications of thin non-rigid unsintered PTFE tape which is inert, sterilizable by heat, non-stick, an effective barrier to moisture and chemicals, and is opaque. PMID- 8405614 TI - Wegener's granulomatosis: a case study in general dental practice. AB - Wegener's granulomatosis is a disease involving renal, lung and vascular tissue, but patients also present to the general dental practitioner with intra-oral manifestations. This case study describes the early onset, progression and treatment of the presenting intra-oral manifestations and the periodontal and endodontic implications of Wegener's granulomatosis. PMID- 8405615 TI - Sandblasting and tin-plating-surface treatments to improve bonding with resin cements. AB - The superior cementation strengths of the adhesive resin cements can now be used in the dental surgery for posts, crowns and bridges and for intra-oral repairs to fractured porcelain fused to metal crowns or bridges, thanks to the availability of miniature sandblasters and portable tin-platers. The author describes the techniques involved. PMID- 8405616 TI - Minor oral surgery for children: 1. Medical problems influencing management. AB - This is the first paper in a series which will deal with exodontia and minor oral surgery for child patients. This first article is concerned with medical conditions that influence the management of children; subsequent papers will consider methods of pain control and surgical techniques for young patients. The final article will give an account of our experience in the provision of exodontia and minor oral surgery for children under local anaesthesia and relative analgesia. Suggestions for further reading will be provided with the final paper. PMID- 8405617 TI - Porcelain laminate veneers: a clinical success? AB - The rapid increase in the use of porcelain veneers is a result of the demand for a minimally invasive, colour and contour stable restoration. However, such restorations are not equally successful in all patients. This article describes the careful treatment planning and patient selection necessary to avoid problems and to ensure that the best result is achieved with porcelain veneers. PMID- 8405618 TI - Toothwear histories: a sensitive issue. AB - Wear of the surface of the teeth is a natural consequence of ageing, but various conditions can render it pathological. In this article the authors describe two strategies for obtaining sensitive, personal information from patients who may be reluctant to disclose such details to a stranger. PMID- 8405619 TI - [Regional prevalence of diabetes treated with oral hypoglycemic agents]. AB - The regional prevalence of diabetes, in France, has been studied using data from a nationwide household survey conducted by INSEE and CREDOC, two national demographic statistics institutes, in a representative sample of 7,323 households (21,007 subjects). Each subject reported, on a form, all the drugs bought during the study period. Subjects reporting, at least once, an antidiabetic oral drug were classified as diabetic. Two hundred and sixty six subjects reported having bought one or more oral antidiabetic drugs, leading to an overall prevalence rate of 1.27%. Prevalence rates were calculated by region (range: 0.86% Pays de Loire- 2.77% Franche-Comte) and by large administrative areas (ZEAT) (range: 0.90% Nord Pas-de-Calais--1.92% Est). In both cases, a North-west, South-east gradient appeared, with the lowest rates in the North and West of France. This trend was maintained after standardization according to the age distribution in the regions studied. The rates are similar to other data in the literature, in particular those derived from national drug sales data, but there is no explanation for this inter regional variability, which merits specific surveys. PMID- 8405620 TI - Heterogeneity of type 1 diabetes at onset in children: results from the French Incidence Study. AB - OBJECTIVE: The aim of this survey was to identify specific relationships between clinical and biological features and immunogenetic markers at presentation in a large cohort of newly diagnosed Type 1 diabetic children recruited from the French incidence registry, in which country the incidence rate of the disease is low. RESEARCH DESIGN AND METHODS: A prospective study of the incidence of Type 1 diabetes has been set up in four regions of France since 1988 (2.3 million inhabitants under 20 years of age) where the epidemiological characteristics of the children have been studied as well as presentation at diagnosis. RESULTS: Five hundred and twenty one cases of newly diagnosed Type 1 diabetes were identified over the 3 years of the study in subjects aged 0-19 yr. The mean incidence rate was 7.6/10(5) per year. Eighty-two percent of the children had a symptomatic period shorter than or equal to 2 months. The mean weight loss was 9.2 +/- 7% of body weight. Forty-eight percent of the children demonstrated plasma total CO2 values < or = 18 mmol/l. Islet cell antibodies and insulin auto antibodies were found in 86% and 41% of the children, respectively. Insulin auto antibody positivity was significantly more frequent in the younger age group (0-4 yr: 78%, p = 0.0001) but not so for islet cell antibodies. HLA-DR3/DR4 phenotype was found in 32% of the children, and 11% carried none of these antigens, irrespectively of age. DR3 phenotype was significantly associated with a lesser frequency of islet cell antibodies (p < 0.001), but no other immunogenetic marker was associated to any clinical feature studied at presentation. CONCLUSIONS: France is a country with a low incidence rate of Type 1 diabetes within Europe. The heterogeneity of the risk for Type 1 diabetes across this continent has recently been confirmed. The variability of the risk according to age has also been shown in many parts of the world. Young age is an important factor for severity at presentation, but immunogenetics do not seem to play a major role in the heterogeneity of the picture at diagnosis. PMID- 8405621 TI - Regular physical activity and reduced occurrence of microalbuminuria in type 2 diabetic patients. AB - In order to evaluate the influence of regular non-strenuous physical exercise on the appearance of microalbuminuria in Type 2 (non-insulin-dependent) diabetic patients, we have studied a cohort of 372 Type 2 diabetic patients (152 males and 220 females, mean age: 63.59 +/- 0.70 years, evolution time: 10.31 +/- 0.4 years, M +/- SEM). One hundred and ninety seven (52.9%) presented normo-albuminuria, 124 (33.3%) microalbuminuria and 51 (13.7%) proteinuria. These three groups were different with regard to evolution time, weight, BMI, waist-hip ratio, Hb1Ac value, prevalence of hypertension and physical activity level. 132 (35.4%) patients had a regular exercise-induced caloric expenditure under 500 kcal/wk whereas 122 (32.7%) were between 500 and 1.000 kcal/wk and 118 (31.7%) over 1.000 kcal/wk. Prevalence of normo-albuminuric patients was 40.1%, 52.4% and 67.7% respectively (p < 0.01). Prevalence of normo-albuminuric patients remained significatively higher in the patient with the greater physical activity level when adjusted to systolic blood pressure, diastolic blood pressure, evolution time and HbA1c value. Our results stress the fact that regular non-strenuous physical activity may have a protective effect on the appearance of microalbuminuria in Type 2 (non-insulin-dependent) diabetic patients. Whether the cardiovascular protective influence of exercise in these patients depends on such an effect remains unknown. On the basis of this cross-sectional evidence, a longitudinal study is now under way. PMID- 8405622 TI - [The triad obesity, hypertension and glycoregulation complaints in a suburban population of Tunisia's Sahel]. AB - Obesity, hypertension and glucose intolerance are often related, the relationship between hypertension and glycemia is well documented and may be explained through hyperinsulinemia. Most of the studies were conducted in western societies. The simultaneous study of this triad in a random sample (n = 555) of a suburban adult community of Tunisian's Sahel, showed an association between hypertension and level of overweight which was also related to diabetes prevalence after matching for age and sex. However serum glucose was related to both systolic and diastolic blood pressure only for non obese subjects, when levels of overweight are stratified. The association between diabetes and hypertension therefore involves different mechanisms for obese and non obese subjects. PMID- 8405623 TI - Expression of certain antiproliferative and growth-related genes in isolated mouse pancreatic islets: analysis by polymerase chain reaction. AB - With the aim at understanding the mechanism responsible for the low proliferative capacity of the adult pancreatic B-cell, expression of the "antiproliferative" gene retinoblastoma and that coding for p53 as well as certain genes coding for cyclin B1, cyclin D1 and p34cdc2 and p33cdk2 kinases, all of which are important for the cell cycle and mitosis, was assessed by the polymerase chain reaction in a semiquantitative assay. Islet expression of the p53 and retinoblastoma genes was higher than that of the liver, but lower or the same as that of the spleen, making expression of these genes an unlikely explanation for the low replicatory capacity of the B-cell. Similarly, islet expression of the genes coding for p33cdk2 kinase and cyclin D1 was not very different from that of the spleen. The levels of cyclin B1 mRNA and the mRNA coding for the p34cdc2 kinase were however both low in islets, compared with those of the spleen and the insulin-producing RINm5F cell line. Stimulation of B-cell proliferation did not change the expression of any of the genes studied. It is concluded that the low expression of cyclin B1 and p34cdc2 kinase and failure to induce these by stimulation of B cell replication may play a role in maintaining adult B-cells in a state of low proliferative capacity. PMID- 8405624 TI - [Standardization of fructosamine assay]. PMID- 8405625 TI - Genetic counselling in prenatal diagnosis. PMID- 8405626 TI - Tramadol in labour pain in primiparous patients. A prospective comparative clinical trial. AB - The analgesic efficacy and safety of tramadol 50 mg, 100 mg and pethidine 75 mg, administered intramuscularly were compared in a randomized, double-blind clinical trial in 90 pregnant women with labour pain. Pain relief was measured by a 4 point verbal rating scale 10, 20, 30, 45 and 60 min after the administration of study drugs. The average total pain relief score within the first hour was 0.9 with tramadol 50 mg, 1.7 with tramadol 100 mg and 1.7 with pethidine 75 mg. In comparison to both tramadol doses the administration of pethidine was associated with a significantly higher frequency of adverse events and a significantly lower respiratory rate in the neonates. The results indicate that tramadol 100 mg is as effective as pethidine 75 mg but has a superior safety profile. PMID- 8405627 TI - Bone mineralisation, body fat and anthropometric measurement in mothers delivering preterm. AB - Twenty mothers of preterm babies who had survived to 1 year old, were matched for age and parity of the mother and time of birth of the baby, with 20 mothers delivering fullterm. Bone mineral, body composition and anthropometric measurements were obtained for each mother and analysed using paired t-tests. The only significant difference (P < 0.01) between the groups was a lower fat-free mass in the preterm mothers calculated from skinfold thickness measurements. PMID- 8405628 TI - Update on epidural analgesia during labor and delivery. AB - Properly administered epidural analgesia provides adequate pain relief during labor and delivery, shortens the first stage of labor, avoids adverse effects of narcotics, hypnotics, or inhalation drugs and it could be used as anesthesia in case a cesarean section is required. Epidural analgesia should be provided to all patients who need and ask for it with an exception of contraindications such as coagulation disorders, suspected infection or gross anatomic abnormality. The technique must be carried out with care if serious life-threatening complications, such as intravenous or intrathecal injection of local anesthetic, are to be avoided. The aim of many recent investigations has been to reduce the total dose of local anesthetic used. Supplementation of an opioid (mainly fentanyl) and introduction of the patient controlled epidural pump may not only serve this goal, but also reduce the demands on the time of obstetric anesthetists. We conclude that properly and skillfully administered epidural is the best form of pain relief during labor and delivery and we hope that more mothers could enjoy its benefits. PMID- 8405629 TI - Doppler velocimetry and umbilical cord blood gas assessment of twins. AB - In 25 pairs of twins delivered by cesarean section, the flow velocity waveform in the umbilical artery was measured and the resistance index calculated. Umbilical artery and vein blood gases were measured at delivery. Results were tabulated by taking the difference in resistance indices between each fetus and plotting these values against the pO2-differences. The data revealed that index differences of more than 10 were of little value in detecting a pO2-difference of > 1 kPa, despite a sensitivity of 100%. The predictive value was only 41%, and specificity 44%. It was obvious, however, that a pO2-difference > 1 kPa occurred only when the resistance index was 76% or more in one fetus and below 76% in the other. The sensitivity of this index parameter was 100%, predictive value 64%, and specificity 78%. In conclusion, the pO2-level in a fetus seems to be unaffected by the impedance in umbilical circulation as long as the resistance index is below 76%. Above this value, there is a significant risk for fetal hypoxaemia. PMID- 8405630 TI - A high uterine artery pulsatility index reflects a defective development of placental bed spiral arteries in pregnancies complicated by hypertension and fetal growth retardation. AB - INTRODUCTION: The development of PIH is associated with a defective trophoblast invasion and conversion of spiral arteries into low-resistance uteroplacental arteries. Hypertension may then be a compensatory response to a defective uteroplacental perfusion. Similar mechanisms may operate in IUGR. AIM: To compare uterine artery Doppler blood flow measurements with placental bed histology. The hypothesis was that placental bed vessel pathology plays a role for a raised flow resistance. MATERIALS AND METHODS: After blood flow measurements, a placental bed biopsy was taken at CS in 26 complicated (study group) and 29 uncomplicated pregnancies (control group). RESULTS: The uterine artery PI was significantly more often abnormally high in the study group compared with the control group, and also in hypertensive pregnancies compared with normotensive IUGR pregnancies. Physiological vessel changes were found in all controls but were absent in 76% of study cases. Physiological changes were significantly more often absent in SGA than in AGA newborns. Absence of physiological changes were significantly more often found in cases with an abnormally high PI. DISCUSSION: The results link together circulatory and structural pathophysiological changes of the uteroplacental unit. A defective physiological conversion of the spiral arteries was associated with an increased uterine flow resistance. CONCLUSION: This study gave further support for the existence of a triad of defective placental bed vessel maturation, increased uteroplacental flow resistance, and hypertension. PMID- 8405631 TI - Effect of progestogens on estrogen-induced lipoprotein changes. AB - A study was performed to evaluate the role of progestogens, on estrogen-induced changes in lipoprotein levels. Sixty postmenopausal symptomatic women, aged 36 59, were included in the study. They were prospectively randomized to a sequential schedule (n = 20), 17 beta-estradiol transdermally 0.05 mg/day on days 1-24 and medroxyprogesterone acetate 10 mg/day orally on days 15-24 or a continuous schedule (n = 20), 17 beta-estradiol transdermally 0.05 mg/day and medroxyprogesterone acetate 2.5 mg/day orally on days 1-24. Patients who had total abdominal hysterectomy+bilateral salphingooopherectomy (n = 20) received only 17 beta-estradiol 0.05 mg/day continuously. Serum total cholesterol (TC), high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol and triglyceride (TG) levels were determined prior to and at the 3rd, 6th and 9th month of therapy in all groups. Mean TC, TG and LDL cholesterol levels did not change significantly during therapy (P > 0.05). Only the mean HDL cholesterol levels showed significant increases in all groups; from 42.30 +/- 9.97 mg/dl to 64.10 +/- 6.81 mg/dl in group I (P < 0.001), from 41.85 +/- 9.09 mg/dl to 60.65 +/- 7.41 mg/dl in group II (P < 0.001) and from 40.70 +/- 11.26 mg/dl to 58.80 +/- 7.74 mg/dl in group III (P < 0.001). It is concluded that medroxyprogesterone acetate, whether used continuously or sequentially, does not oppose the beneficial effects of transdermal 17 beta-estradiol on the lipoprotein profile. PMID- 8405632 TI - Ultrasonographically guided fine needle aspiration cytology and core-needle biopsy in the diagnosis of breast tumors. AB - The present study was designed to assess the performance of fine needle aspiration cytology (FNAC) and core-needle biopsy (CNB) of breast lesions when these procedures are performed under sonographic guidance. The results obtained in 1142 FNAC procedures and 180 CNB procedures were analysed. The study took place in a University Hospital and a private practice clinic. The patients eligible for this study were a series of women, in whom at least one hypoechoic, limited mass was found at breast ultrasonography. Cystic masses were excluded from the study. Each individual mass was investigated using either FNAC or CNB under sonographic guidance. Accuracy characteristics to suspect or diagnose malignant and pre-malignant breast lesions, such as sensitivity and specificity, were calculated. The cytological results were classified into four categories according to standard criteria: benign; atypical and/or suspicious for cancer (hyperplasia with atypia); malignant; and unsatisfactory for diagnosis specimen. Tissue specimens were classified according to the W.H.O. The 1142 lesions submitted to FNAC included 66 invasive carcinomas, 4 intraductal carcinomas, and 4 atypical hyperplasias. FNAC led to 6 false-negative examinations, equally distributed between small (volume < or = 1 cm3), and larger lesions, and 1 false positive examination. There were 12.9% (147/1142) inadequate smears. Only 3 inadequate samples were obtained in the presence of a discrete pathologic lesion (3/363, or 0.1%). All 3 corresponded to invasive carcinomas. The majority of inadequate samples (144/147, or 98.0%) were obtained in the normal/dystrophic group. Overall, the sensitivity is 92.1%, and the specificity is 84.8%. The 180 lesions submitted to CNB included 31 invasive carcinoma, 5 intraductal carcinomas, and 17 atypical hyperplasias. CNB, in this series, had an accuracy rate of 100%. In conclusion, US guidance increases the accuracy of breast tissue sampling procedures. This is of particular importance as the number of suspicious images to be investigated steadily increases, as the result of mass screening. PMID- 8405633 TI - Female urinary stress incontinence in terms of connective tissue biochemistry. AB - The role of connective tissue in the aetiology of female stress incontinence has been investigated. Collagen content and extractability as well as estrogen receptor concentration in vesico-vaginal fascia were measured after small tissue biopsies had been obtained during vaginal repair surgery in cases of urinary incontinence. The mean concentration of estrogen receptor in vesico-vaginal fascia among incontinent women was 49.4 +/- 14.8 fmol/mg of protein as compared to 29.6 +/- 13.1 in continent control group (P < 0.03; t-test). The mean hydroxyproline concentration in vesico-vaginal fascia of incontinent women was 13.8 +/- 2.6 micrograms/mg wet weight, whereas in the control group it was significantly higher 20.6 +/- 2.4 (P < 0.001). The role of connective tissue components in the aetiology of female stress incontinence is discussed. PMID- 8405634 TI - Angiosarcoma of the uterus following radiotherapy for squamous cell carcinoma of the cervix. AB - A case report of a 61-year-old woman who developed a uterine angiosarcoma 6.5 years after primary radiotherapy for a moderately well differentiated squamous cell carcinoma of the uterine cervix stage IB is presented. The diagnosis was confirmed by immunohistochemical studies showing positive staining for Ulex europaeus agglutinin 1 and supported by ultrastructural studies. Though various post-irradiation angiosarcomas are reported in the literature, this is, to our knowledge, the first report of an angiosarcoma of the uterus after radiotherapy. PMID- 8405635 TI - Abnormal cerebral hemodynamics in pregnancy-related hypertensive encephalopathy. AB - A patient with an imminent pregnancy-related hypertensive encephalopathy is presented in whom transcranial Doppler ultrasound studies revealed increased middle cerebral artery blood flow velocities before and up to 7 days after cesarean section. These increased velocities are normally caused by cerebral vasospasm and subsequent hypoperfusion in pregnancy-related hypertensive encephalopathy. Additional duplex volume flow studies of the common carotid artery were in favor of cerebral hyperperfusion as a cause of the increased blood flow velocities. The clinical implications of these findings are discussed. PMID- 8405636 TI - Postpartum hemolytic uremic syndrome following placental abruption. AB - Hemolytic uremic syndrome associated with pregnancy is a rare condition. Authors report a patient treated with corticosteroids for bronchial asthma who was afflicted by placental abruption at 24 weeks' gestation. The abruption was preceded by developing herpes zoster and by deteriorating respiratory symptoms. The induced labor was followed by anuria, acute renal failure, microangiopathic hemolytic anemia, thrombocytopenia then fever and hypertension. The patient was treated early with plasma infusion, transfusion and hemodialysis. She recovered completely after 7 weeks. This case seems to be unique inasmuch as the hemolytic uremic syndrome was preceded by prodromal illness during pregnancy and was associated with placental abruption. PMID- 8405637 TI - Intravenous sulprostone and uterine scarring based upon 22 cases of therapeutic abortion during 2nd and 3rd trimesters of pregnancy. AB - A preliminary study in 22 patients with uterine scarring was undertaken using sulprostone by intravenous infusion when therapeutic abortion was deemed necessary during the 2nd and 3rd trimesters of pregnancy. The dosage used was 500 micrograms by slow infusion lasting 10 h. There were no cases of ruptured uterus. Adverse reactions were absent. Results were satisfactory. Mean induction expulsion duration: 11 h. Expulsion rate in 24 h: 63%. With strict monitoring and in a specialized center, this technique may be suggested when a late therapeutic abortion with a scarred uterus is indicated. PMID- 8405638 TI - Labetalol therapy in pregnancy induced hypertension: the effects on fetoplacental circulation and fetal outcome. AB - We prospectively studied the effects of oral labetalol therapy in patients with moderate to severe pregnancy induced hypertension (PIH). The outcome variables were blood pressure control, effect on umbilical artery flow velocity waveforms (UAFVW) and fetal outcome. Forty-two patients were recruited, all had moderate to severe PIH. The mean systolic blood pressure (SBP) and diastolic blood pressure (DBP) on entry were 154 +/- 7 mmHg and 104 +/- 5 mmHg, respectively. All had significant proteinuria. After 1 week on labetalol therapy, 85% of patients had their blood pressure controlled. The reduction in both SBP and DBP was statistically significant. There were no significant changes in UAFVW, Resistance Index (RI), uric acid or platelets. The mean fetal age on entry was 246 +/- 10 days while gestation at delivery was 258 +/- 17 days. The mean birth weight was 2712 +/- 609 g. No perinatal mortality occurred in this study. Labetalol is an effective drug in controlling blood pressure and does not adversely affect the UAFVW. No neonatal problems were attributed directly to the drug. Fetal outcome was satisfactory despite the 12 fetuses that were growth-retarded. Labetalol allows safe prolongation of pregnancies complicated by PIH. PMID- 8405639 TI - Maternal characteristics and expected birth weight. AB - Fetal growth charts currently used aggregate birth weights of infants with various natural histories from 1931 until 1967. In order to modernize these charts, avoiding deviation from the natural history of fetal development, we report data from infants born after spontaneous onset of labour in 'normal' pregnancy from a gestational age of 267 to 295 days between 1972 and 1982 (n = 14,113). The relationship between birth weight and gestational age in days was studied by multiple regression analysis, containing dummy variables for parity and gender. The estimated proportion of the variance in the model, attributed to these characteristics, was 15%. This could be improved to 22% by supplementing the model with maternal characteristics such as age, height, mid-pregnancy weight and ethnic origin. According to this extended model, in the Dutch section of the population 511 (4.6%) babies had a birth weight below the 5th percentile, whereas 412 (3.7%) babies would be labeled as such according to the conventional birth weight tables. Moreover, 93 babies would be wrongly considered too small, corresponding with a sensitivity of 62.4%, and 192 babies would be wrongly considered normal, corresponding with a specificity of 99.3%. Integration of the four currently used tables into one, and adjustment for easily available maternal characteristics, could substantially improve classification methods. PMID- 8405640 TI - Ovulation induction in polycystic ovary syndrome: a review of conservative and new treatment modalities. AB - Polycystic ovary syndrome (PCOS) is a complex disorder with heterogeneous clinical and endocrine features. Chronic anovulation and infertility are common and affect about 75% of the patients. Ovulation induction in PCOS patients is a challenge for the physicians who treat these patients. Several different treatment modalities have been proposed to induce ovulation in PCOS, each of which deals with a different clinical or endocrine disorder which is present in these patients. This review presents traditional and new methods for ovulation induction in PCOS patients, the theoretical background, the pros and cons for each treatment and success rate. PMID- 8405641 TI - Surgical support and suspension of genital prolapse, including preservation of the uterus, using the Gore-Tex soft tissue patch (a preliminary report). AB - Abdominal-retroperitoneal sacral genito-colpopexy using the expanded polytetrafluoroethylene (ePTFE) soft tissue patch has been found to be highly effective for repair of genito-vaginal prolapse. We treated 61 patients in this way, including patients who had failed multiple previous attempts at repair. At a mean of 32 months of follow-up, more than 95% of patients were still classified as successfully treated. To preserve the uterus in cases of complete genito vaginal prolapse, we have developed a new surgical technique, which we describe in this paper. PMID- 8405642 TI - A prospective randomized study comparing amoxycillin/clavulanate with cefuroxime plus metronidazole for perioperative prophylaxis in gynaecological surgery. AB - A prospective, randomized, open study was performed in 199 patients at the Leyenburg Hospital comparing amoxycillin/clavulanate (AMX/CL) with cefuroxime plus metronidazole (CR/MN) in the prophylaxis of infection following gynaecological surgery. AMX/CL was given as a single dose of 2200 mg i.v. at the start of the operation. CR/MN, 750/500 mg i.v. was administered 3 times within 24 h, beginning at the start of the operation. The study group consisted of patients undergoing either a vaginal hysterectomy, a vaginal hysterectomy with cysto/rectocele repair or a secondary caesarean section. There were no statistically significant differences in demographic characteristics, duration of surgery or anaesthetic method between the two groups. Postoperatively, 10.6% of patients developed a urinary tract infection, and febrile temperatures were found in 9.0% of patients. There were no statistically significant differences between the two treatment groups. Other complications were found in less than 1% of the study population, equally distributed between the two regimens. In this study there was a low overall percentage of infection after gynaecological surgery. AMX/CL was as effective as CR/MN as a perioperative prophylactic treatment and has the dual advantage of a single dose and lower cost. PMID- 8405643 TI - Dutch general practitioners' attitudes towards the climacteric and its treatment. AB - In order to obtain insight into general practitioners' (GPs) point of view regarding climacteric problems and possible treatment, a survey was conducted among 195 GPs in the Limburg Province in the Netherlands. Ultimately, 159 GPs participated (response-rate 82%). The GPs were either members of the Society for the Improvement of the Expertise of GPs, or filled out the questionnaire previous to participation in a medical training course. The climacteric and its typically related complaints were generally well recognized by the surveyed GPs. Ninety five percent of the GPs were of the opinion that women with climacteric complaints need some kind of medical help, and 62% were found to prescribe medication, mainly hormone replacement therapy (HRT). Remarkably, 75% of all GPs were of the opinion that prescribing HRT for climacteric complaints is a form of medicalization. Twelve percent of the GPs stated they never prescribe HRT for solely preventive purposes. In case of preventive treatment merely 32% of the GPs prescribed HRT for a period longer than 5 years. PMID- 8405644 TI - A comparison of pefloxacin/metronidazole and doxycycline/metronidazole in the treatment of laparoscopically confirmed acute pelvic inflammatory disease. AB - A double-blind, randomised study was conducted to compare the efficacy and safety of a combination of pefloxacin and metronidazole versus doxycycline and metronidazole in patients with pelvic inflammatory disease (PID). The clinical diagnosis had to be confirmed by laparoscopy before patients were included. Of the 74 patients who fulfilled the clinical criteria for PID, laparoscopy confirmed the diagnosis in only 40 patients (54%). The microorganism most frequently found as causative pathogen was Chlamydia trachomatis. Both treatment groups showed a good response to the study-medication. At discharge 9 patients in the pefloxacin group (45%) were cured and 10 patients (50%) had improved. In the doxycycline group 7 patients (35%) were cured and 10 patients (50%) had improved. Obviously pefloxacin/metronidazole and doxycycline/metronidazole are equally effective in the treatment of PID. PMID- 8405645 TI - Diagnosis of Fitz-Hugh-Curtis syndrome by ultrasound. AB - Chlamydiae are often responsible for the development of perihepatitis and/or diffuse peritonitis after salpingitis in sexually active women, the so-called Fitz-Hugh-Curtis syndrome (FHC). The definitive diagnosis can be made by direct culture, serological evidence and laparoscopy/laparotomy. Violin-string adhesions between the liver surface and the abdominal wall can be seen. Two patients are described in whom the diagnosis was made by ultrasound: the violin strings and ascites were clearly seen. Serological and operative evidence verified the diagnosis. CONCLUSION: ultrasound may become an extra tool to diagnose the FHC syndrome in an early and non-invasive way. PMID- 8405646 TI - Epidemiological and birth weight characteristics of triplets: a study from the Dutch twin register. AB - From 112 triplet sets, born in The Netherlands from the end of 1986 to the beginning of 1991 and registered in the Dutch Twin Register, several details such as birth weight, gestational age, zygosity, and etiology were assessed by questionnaire, which was filled out by the mother. For 33 triplet sets, zygosity was also assessed by blood typing. Maternal smoking during pregnancy was also noted. Results show a very strong increase in number of triplets caused by artificial fertility enhancing techniques and consequently a shift in the relative contribution of zygosity types to the total number of triplets. Birth weight is predominantly influenced by gestational age. Other effects on birth weight are controlled for possible confounding with gestational age. First born triplets weigh more than later born triplets; boys weigh more than girls; nearly 25% of all individual triplets weigh less than 1500 g, i.e. belong to the category very low birth weight (VLBW); regular maternal smoking produces a 14% birth weight reduction; ovulation induction seems to decrease the sex ratio, i.e. hormonal treatment with ovulation inducing substances increases the probability of female offspring. PMID- 8405647 TI - Birth weight in opposite sex twins as compared to same sex dizygotic twins. AB - The question addressed in the present report is whether the large birth weight differences in dizygotic twin pairs of opposite sex (DZos), especially in 'male first' couples--observed by Blickstein and Weissman (Blickstein I, Weissman A. Birth weight discordancy in male-first and female-first pairs of unlike-sexed twins. Am J Obstet Gynecol 1990;162:661-663) and replicated in the present study- can be explained by two general influences on birth weight, viz. sex and birth order, or whether some specific effect on fetal growth has to be assumed that is present only in twin pairs of differing sex. The associated enhanced health risk would hit the female twin (from a male first-female second couple) in the first place. If the hypothesis is correct, then one may expect that birth weight of twins is somehow dependent on the sex of the co-twin. This was studied in 3069 twin pairs born in The Netherlands since the end of 1986. Results show that among DZ twins, birth weight is not affected by the sex of the co-twin. Therefore, birth weight differences in DZos pairs have to be ascribed to the general effects of sex and birth order. There is no effect that is specific to DZos pairs only. PMID- 8405648 TI - Therapeutic abortion during the 2nd and 3rd trimesters of pregnancy using a synthetic derivative of prostaglandin E2, sulprostone, administrated intravenously. Based upon 182 cases. AB - OBJECTIVE: The purpose of the study is to assess the efficacy of and adverse events linked to the use of intravenous sulprostone during the 2nd or 3rd trimesters of pregnancy for therapeutic abortion. STUDY DESIGN: One hundred eighty-two patients (70 nulliparous, 112 multiparous) were hospitalized for therapeutic abortion. The route of administration was invariably intravenous and one single dose of sulprostone was used: 1000 micrograms of sulprostone diluted in 1 l of isotonic saline solution given as a 10-h infusion. RESULT: Expulsion within the first 24 h was obtained in 70% of cases with a mean induction expulsion interval of 14 h. In three cases, laparotomy was required for hemorrhagic syndromes. CONCLUSION: Intravenous sulprostone enable evacuation of uterine contents with minimal adverse reaction. Attention should nevertheless be drawn to the existence of hemorrhagic syndromes. PMID- 8405649 TI - Mechanism of insulin action, role of ions and the cytoskeleton. PMID- 8405650 TI - Does carnosine possess direct antioxidant activity? AB - 1. A brief review of development of ideas of the antioxidant activity of carnosine and related compounds is presented. 2. An analysis of the behaviour of carnosine in different models of free radical chain reactions shows that carnosine is a potent hydrophilic antioxidant of a direct non-enzymatic action. 3. It is characteristic of the higher activity of interaction with active hydroxyl radical. 4. However the known biological effects of carnosine cannot be explained only by its anti-oxidant properties. PMID- 8405651 TI - Modulation of the ATP induced [Ca2+]c increase in AS-30D hepatoma cells. AB - 1. The regulation of the increase in the cytosolic calcium concentration ([Ca2+]c) induced by extracellular ATP in AS-30D hepatoma cells was studied. 2. Homologous desensitization involving the refilling of intracellular calcium pools and the participation of protein kinase C was found. 3. Isoproterenol, forskolin and dibutyryl-cyclic AMP also induced an increase in [Ca2+]c. 4. Interestingly, synergism was found for isoproterenol or forskolin and ATP. 5. The results suggest that there are two pathways for mobilizing [Ca2+]c in AS-30D hepatoma cells; one is activated by ATP receptors and the other by cyclic AMP. PMID- 8405652 TI - Inhibition kinetics of brain butyrylcholinesterase by Cd2+ and Zn2+, Ca2+ or Mg2+ reactivates the inhibited enzyme. AB - 1. The inhibition kinetics of sheep brain butyrylcholinesterase (BChE) (acylcholine acylhydrolase, EC 3.1.1.8) by Cd2+ and Zn2+ has been studied. 2. KS has been determined as 0.14 mM. Cd2+ and Zn2+ were the hyperbolic mixed-type inhibitors of BChE. Ca2+ and Mg2+ had no effect on the enzyme activity in the experimental conditions. 3. But when the enzyme was inhibited by 0.1 mM Cd2+ or Zn2+, Ca2+ and Mg2+ reactivated the inhibited form of BChE. PMID- 8405653 TI - Distribution of beta-adrenergic binding in fractionated porcine adipocytes. AB - 1. The subcellular distribution of the porcine adipocyte beta-adrenergic receptor was studied in fractionated adipocytes. 2. The 30,000 g pellet obtained from hypotonically lysed cells contained membrane vesicles and mitochondria; it yielded approx 200-300 fmol dihydroalprenolol-bound receptors/mg protein. 3. Activity was increased to about 1000 fmol/mg protein after isolation of a plasma membrane fraction on a Percoll gradient. 4. The 5'-nucleotidase, succinate dehydrogenase and lactate dehydrogenase activities were usually enriched in compartments different from the ligand-binding activity. 5. Activity of porcine adipocyte 5'-nucleotidase, a purported plasma membrane marker enzyme, was not distributed in the same manner as the beta-adrenergic receptor. PMID- 8405654 TI - Regulation by progesterone and pregnenolone of dimeric aldehyde dehydrogenase from rat testis cytoplasm. AB - 1. Aldehyde dehydrogenase from rat testis cytosol has been purified to electrophoretic homogeneity. With an isoelectric point of 9.5, the enzyme appears a dimer with a subunit molecular weight of 52,500. 2. The influence of pregnenolone and progesterone on the kinetic behaviour has been investigated using valeraldehyde as substrate. 3. The kinetic data were fitted to a modified version of the Monod-Wyman-Changeux model and the fitting procedure resulted in a good correspondence between theoretical and experimental reaction rates over a wide range of valeraldehyde concentrations. 4. According to the model, the dimeric enzyme is in equilibrium between two conformational states R and T. The R state displays higher affinity for valeraldehyde, but lower catalytic power. In the absence of substrates and effectors the [T]/[R] ratio is near to 1. 5. Pregnenolone and progesterone activate the enzyme by stabilizing the more active state T and by increasing the catalytic power of the R state. The increase of activity is counteracted by the inhibition exerted by both steroids on the T state. PMID- 8405655 TI - Metabolic capacity, fibre type area and capillarization of rat plantaris muscle. Effects of age, overload and training and relationship with fatigue resistance. AB - 1. The influences of age (5, 13 and 25-month-old rats), overload as obtained by denervation of synergists, and training on the metabolic capacity, relative muscle cross-sectional area occupied by each fibre type, capillarization and fatigue resistance of the rat m. plantaris were investigated. 2. Creatine kinase, phosphorylase and citrate synthase activities were lower in muscles of 25 than in those of 13-month-old rats (P < 0.001). 3. Overload resulted in an increased relative area of type I and IIa fibres at all ages (P = 0.001). 4. Capillary density decreased with overload and increasing age (P < 0.001). 5. Fatigue resistance was higher in muscles of 13 than in those of 5-month-old rats (P < 0.05), and increased with overload (P < 0.05) at all ages. 6. Fatigue resistance of the whole muscle was not closely related to its oxidative capacity in contrast to what is generally found for single fibres or motor units. PMID- 8405656 TI - Further studies on the mechanism of action of gossypol on mitochondrial membrane. AB - 1. In order to explore the mechanism of inhibition of hydroxylases involved in steroidogenesis, by gossypol, we studied the effect of this drug on adrenal cortex mitochondria, and compared it with those on kidney and heart. 2. The uncoupler effect of gossypol (collapse of delta psi and Ca2+ efflux) was found to be lower in adrenal cortex mitochondria than in kidney and heart mitochondria. 3. Gossypol produced more extensive changes on the membrane lipidic matrix (increase in the order parameter for 5-doxylstearic acid) in adrenal cortex mitochondria than in the other mitochondria studied. 4. The results described above indicate that the mechanism of inhibition of gossypol of steroidogenic adrenal enzymes could be attributed to an alteration of the lipidic matrix which, in turn, modifies protein function. PMID- 8405657 TI - A peptide inhibitor for S-adenosyl-L-methionine-dependent transmethylation reactions. Purification and characterization. AB - 1. A. proteinaceous inhibitor for S-adenosyl-L-methionine (AdoMet)-dependent transmethylation reactions has been purified to apparent homogeneity from rat liver cytosolic fraction. 2. The peptide was made up of 29 amino acid residues with a molecular weight of 2,584. Glycine accounted for 52% of the total amino acids. 3. Employing AdoMet: protein-carboxyl O-methyltransferase (Protein methylase II) and bovine serum gamma-globulin as in vitro substrate, the mode of inhibition was found to be non-competitive with Ki value of 1.9 x 10(-8) M. 4. When the inhibitor was present in the reaction mixture together with S-adenosyl-L homocysteine (AdoHcy), which is a competitive inhibitor for AdoMet, the extent of inhibition exceeded that exerted by each individual inhibitor alone, suggesting that the sites of the inhibitors on the enzyme molecule are different. 5. Almost a stoichiometric relationship exists between the enzyme and the inhibitor molecule, the ratio being approx one. PMID- 8405658 TI - Conversion of a NADPH-dependent aldehyde reducing enzyme into aldose reductase. AB - 1. Aldose reductase, aldehyde reductase and high-Km aldose reductase were purified from the inner medulla of dog kidney. 2. Compared with aldose reductase, high-Km aldose reductase had a lower isoelectric point, a lower activity for aldo sugars and a lower sensitivity for aldose reductase inhibitors, and it was not activated by sulfate ions. Both reductases had the same molecular weight (38,500) and immunochemical properties. 3. High-Km aldose reductase was easily converted into an aldose reductase-like enzyme, namely a generated reductase upon incubation in neutral buffer solution. 4. The generated reductase was identical with aldose reductase with respect to the isoelectric point, substrate specificity, activation by sulfate ions and IC50 values for aldose reductase inhibitors. The generated reductase revealed immunochemical identity with aldose reductase as well as high-Km aldose reductase. PMID- 8405659 TI - Oxidation of 6-hydroxydopamine catalyzed by tyrosinase. AB - 1. The oxidation of 3,4-dihydroxyphenylethylamine (dopamine) by O2 catalyzed by tyrosinase yields 4-(2-aminoethyl)-1,2-benzoquinone, with its amino group protonated (o-dopaminequinone-H+), which evolves non-enzymatically through two branches or sequences of reactions, whose respective operations are determined by the pH of the medium. 2. The cyclization branch of o-dopaminequinone-H+ takes place in the entire range of pH and is the only significant branch at pH > or = 6. 3. The hydroxylation branch of o-dopaminequinone-H+ only operates significantly at pH < 6, and involves the accumulation of 2,4,5 trihydroxyphenylethylamine (6-hydroxydopamine), identified by high performance liquid chromatography (HPLC). 4. 6-hydroxydopamine is also a substrate of tyrosinase. The identification and evolution of the oxidation products of 6 hydroxydopamine has been carried out by spectrophotometry and HPLC assays. PMID- 8405660 TI - Phosphatidylethanolamine: ceramide-ethanolaminephosphotransferase activity in synaptic plasma membrane vesicles. Influence of some cations and phospholipid environment on transferase activity. Further proof of its location. AB - 1. Synaptic plasma membrane vesicles (SPMV) from rat brain synthesized ceramide phosphoethanolamine (SpE), an analogue of sphingomyelin (SpC) from phosphatidylethanolamine (PE) and ceramide. 2. This reaction was catalyzed by PE: ceramide-phosphotransferase. 3. The presence of PC did not modify the SpE synthesis and PI and PS at twice PE concentration seemed to be activators; only PG was an inhibitor at all concentrations. 4. Some cations (Mg2+, Mn2+) were without effect, while Ca2+ increased transferase activity, so was interesting to study. 5. Transferase was compared with sialidase (external enzyme). 6. Kinetics other than those already performed by us were undertaken in order to confirm its location. PMID- 8405661 TI - Oxidation of methionine residues in equine growth hormone by Chloramine-T. AB - 1. Reactivity of methionine residues towards Chloramine-T was studied in the equine growth hormone. 2. With a 20.0-fold molar excess of reagent over methionine, full oxidation of the four residues of the protein is achieved. 3. Methionine 4 is the most reactive group, followed by methionines 72 and 178- methionine 123 being the less reactive residue. 4. As judged by circular dichroism spectra and binding assays, protein conformation and binding capacity to specific receptors remains unchanged even after full oxidation of all four methionine residues. 5. Results agree with data previously obtained with bovine growth hormone. PMID- 8405662 TI - Identification of the major annexins in Ehrlich ascites tumor cells. AB - 1. Three calcium-binding proteins have been purified from Ehrlich ascites tumor cells. 2. They were identified by amino acid sequence analysis on selected fragments obtained by tryptic digestion. 3. The proteins belong to the annexin family and were identified as annexins II, III and V. 4. Antibodies raised against the proteins were used to examine for their presence in a number of murine tissues. 5. The occurrence was found to be in reasonable accordance with earlier reports. PMID- 8405663 TI - Effects of adenine nucleotides on low Km 5' nucleotidase from human seminal plasma. AB - 1. Low Km 5' nucleotidase purified from human seminal plasma has been used in this study to investigate the response of the enzyme ot adenine nucleoside di- and triphosphates in the presence of AMP and IMP as substrates. 2. In the presence of AMP, the addition of 0.5 mM ATP to the enzyme Mg-free results into the highest Vmax/Km ratio value and other experimental combinations of effectors tested cause variation of the kinetic parameters of the enzyme, indicating a control of AMP dephosphorylation by adenine nucleotides. 3. In the presence of IMP, ATP and ADP activate the enzyme but the response to various experimental combinations of effectors shows no significant difference in the kinetic properties of the enzyme, indicating a different control of the dephosphorylation of IMP. PMID- 8405664 TI - Treatment of lactating rats with PCBs induces CYP1A1 and enhances the formation of BP 7,8-dihydrodiol, the proximate carcinogen of benzo(a)pyrene. AB - 1. Treatment of pregnant and lactating rats with a single i.p. dose of 250 mg/kg body weight produced 5-fold and 2-fold increases, respectively, in hepatic P-450 concentrations using microsomes isolated from pregnant, neonatal, lactating and foetal rats. 2. Concomitantly, 26-fold, 20-fold and 14-fold increases in neonatal, maternal and foetal ethoxyresorufin-O-deethylation (EROD), respectively were found, but only 2.5-fold increases could be determined using microsomes isolated from lactating rats. 3. The metabolism of [3H]benzo(a)pyrene was increased 9-fold and 2-fold in pregnant and foetal rats, respectively, but only 2 fold increases were measured for lactating rats. 4. Western blot analysis of microsomal proteins obtained from lactating rats showed significant CYP1A1 and CYP1A2 induction and for the same hepatic tissue 62-fold and 44-fold increases in cDNA hybridized CYP1A1 and CYP1A2 mRNA, respectively, were found. 5. Treatment of lactating rats with PCBs resulted in enhanced formation of all BP-metabolites, but the ratio of diol to total BP-metabolites was more than 3-fold greater. 6. The formation of the proximate carcinogen BP-7,8-dihydrodiol was 5-fold increased and a similar 3-fold increase in epoxide hydrolase activity was estimated for lactating rats. 7. The results of the present study indicates that lactation protects, in part, against the inductive effect of PCBs, possibly by enhanced clearance of these chemicals via lactation. PMID- 8405665 TI - The homeotic transformation of tails into limbs in Rana temporaria by retinoids. AB - The most remarkable of all the effects of retinoids on embryonic systems is the homeotic transformation of tails into legs which was recently reported using an Indian species of frog. Since then several attempts have been made to repeat these results on other species, notably Xenopus, with no success. Here I report the successful repetition of this homeotic transformation using Rana temporaria tadpoles treated with retinyl palmitate. The phenomenon is concentration dependent, time-dependent, and stage-dependent. There is some difference in effect according to the tail amputation level. The limbs induced are always hindlimbs and there can be between 1 and 9 of them. There is a tendency to induce limbs in pairs so that even numbered groups are produced in considerable excess over odd numbered groups. As assessed by cartilage staining the majority of the limbs are normal in the proximodistal and anteroposterior axes. The other types of outgrowths induced are double-posterior limbs, posterior half-limbs (usually defective in the proximodistal axis), and spikes. As assessed by the anatomy of the muscle patterns some of these apparently normal limbs are duplicated in the dorsoventral axis. The stage and species dependencies of this phenomenon suggest that it may involve thyroid hormone receptors whose levels rise leading up to metamorphosis and whose interactions with retinoic acid receptors have recently been described. PMID- 8405666 TI - Midkine, a retinoic acid-inducible growth/differentiation factor: immunochemical evidence for the function and distribution. AB - Midkine (MK) is a heparin-binding growth factor specified by a retinoic acid responsive gene. A rabbit was immunized against recombinant MK produced in L cells, and the resulting antibody was affinity purified using MK-glutathione S transferase fusion protein as a ligand. The MK-specific antibody was used to investigate the function and distribution of MK. MK of the same size as the recombinant MK produced in L cells (13 kDa) was strongly detected in a 13-day rat embryo. Weak expression was observed in the brains of a 19-day embryo, neonates, and adults. In the 13-day mouse embryos, high levels of MK were detected on the surfaces of brain cells, as well as in basement membranes and in the epithelial cells of the intestine, the jaw, and the rib. Nerve cells from the brains of 13 day or 19-day rat embryos extended neurites about twofold more efficiently on MK coated dishes than on poly-L-lysine-coated dishes. Furthermore, anti-MK antibody inhibited neurite extension not only on MK-coated dishes, but also on poly-L lysine-coated dishes. These results suggest that MK is an endogenous neurite outgrowth factor involved in the development of the central nervous system. Anti MK antibody was also found to inhibit growth of Wilms' tumor cells, which secreted MK into culture medium. Thus, overproduction of MK is involved in enhanced growth of Wilms' tumor cells. PMID- 8405667 TI - Preferential expression of alternatively spliced mRNAs encoding type II procollagen with a cysteine-rich amino-propeptide in differentiating cartilage and nonchondrogenic tissues during early mouse development. AB - Type II procollagen mRNAs are alternatively spliced: type IIA mRNA contains an exon encoding a cysteine-rich domain in the amino-propeptide and type IIB mRNA lacks this exon. In mouse embryos between 9.5 and 13.5 days, type IIA mRNA was the major form of Col2a-1 transcript expressed in both prechondrogenic and nonchondrogenic tissues and type IIB mRNAs were present in small amounts. After 12.5 days, type IIB mRNA levels increased rapidly and finally exceeded type IIA mRNAs. Type IIB mRNAs became the major Col2a-1 transcript by 14.5 days, predominantly expressed in maturing chondrocytes. By 17.5 days type IIB mRNAs account for 80% of the Col2a-1 transcripts. Expression of type IIA mRNAs follows the change in the growth pattern of the cartilaginous model of the axial and appendicular skeleton and of the otic capsule and nasal septum. In nonchondrogenic tissues, type IIA mRNAs are more commonly expressed in epithelial structures of ectodermal and endodermal origin than in nonepithelial tissues. The switching of expression from type IIA to type IIB mRNA as major Col2a-1 transcript may be associated with the commitment of precursor cells to the chondrocyte lineage and sites of type IIA mRNA expression may mark regions of potential cartilage growth. The differential expression pattern of type IIA mRNAs therefore points to an association of type IIA procollagen with chondrocyte differentiation during cartilage growth and some function early in embryogenesis in the epithelial organization of nonchondrogenic tissues. PMID- 8405668 TI - The DVR-1 (Vg1) transcript of zebrafish is maternally supplied and distributed throughout the embryo. AB - It is not known how region- or tissue-specific differences are generated in the zebrafish embryo. To look at the potential role of maternal transcripts in generating cell diversity, we have isolated and characterized the zebrafish homologue of Xenopus DVR-1 (Vg1), a maternally supplied RNA that encodes a member of the transforming growth factor-beta superfamily. The zebrafish DVR-1 RNA is maternally supplied and its protein product shares a high degree of sequence identity with Xenopus DVR-1. These conserved features indicate that DVR-1 is likely to have an essential function in early embryogenesis. However, unlike the frog transcript, which is restricted to vegetal cells, DVR-1 RNA is distributed equally among all zebrafish blastomeres. We suggest that the ubiquitous distribution of DVR-1 RNA reflects a significant aspect of the developmental strategy of the zebrafish in which each blastomere retains an equivalent developmental potential throughout the cleavage period. PMID- 8405669 TI - Calcium regulation of neural fold formation: visualization of the actin cytoskeleton in living chick embryos. AB - The involvement of calcium ions during chick neurulation was studied by treating neural plate stage embryos with agonists and antagonists of calcium transport and with inhibitors of calmodulin activity. Organotypic shape changes were examined by light and scanning electron microscopy. Changes in size of cell apices were quantitated by computer image analysis of actin filaments labeled with fluorescent phallicidin in time lapse recordings of living embryos. Both ionomycin and A23187 caused precocious fold elevation around the median hinge point and convergence by bending at the lateral furrows only when Ca2+ was in the external medium. As judged by decreased perimeters of 100 fluorescent apical polygons, cell apices constricted medial to the lateral furrows but did not change significantly within the median hinge point. Pretreatment with dihydrocytochalasin B or cytochalasin D prevented precocious folding and apical constriction. Papaverine and verapamil prevented folding but could be reversed by subsequent ionophore treatment. Calmidazolium and trifluoperazine irreversibly blocked folding. The demonstration in living embryos of constriction of lateral cell apices in a calcium-dependent manner is consistent with a contractile process operating during the formation of neural folds. PMID- 8405670 TI - Ontogenesis of chick iris intrinsic muscles: evidence for a smooth-to-striated muscle transition. AB - The iris sphincter muscle consists of striated muscle fibers in the adult chicken. The ontogenetic development of this muscle has been studied by immunocytochemistry, from Embryonic Day 7 to the time of hatching. The time course of expression of specific markers of either smooth or striated muscle, i.e., smooth and skeletal muscle myosin heavy chains along with the inositol 1,4,5-trisphosphate (IP3) receptor, the intracellular Ca2+ release channel gated by the second messenger IP3, was investigated. We observed the parallel and transient expression, from about Embryonic Day 8 until hatching, of smooth muscle myosin heavy chain and IP3 receptor in early differentiating cells at the pupillary margin, as well as in migrating cells at subsequent stages; the sequential expression of sarcomeric myosin heavy chain around the end of the second embryonic week; and its spreading throughout the iris sphincter by the time of hatching. The present findings are consistent with a smooth muscle differentiation stage as an intermediate stage in the ontogenic development of the iris sphincter muscle of the chicken. PMID- 8405671 TI - Retinoic acid stimulates mouse lung development by a mechanism involving epithelial-mesenchymal interaction and regulation of epidermal growth factor receptors. AB - Retinoic acid (RA) stimulated proliferation of both epithelial and mesenchymal cells in cocultures isolated from developing mouse lungs. There was a corresponding increase in epithelial branching activity in organ culture of embryonic lungs exposed to similar doses of RA. Stimulation was maximal with concentrations of 1 microM and progressively decreased with either lower or higher concentrations. However, when lung cell monocultures of isolated epithelial and mesenchymal cells were exposed to RA, the mitogenic effect was observed only in the mesenchymal population. This suggests that RA may not have a direct mitogenic effect on epithelial cells but rather functions indirectly through the mesenchyme. The cellular response to RA was correlated with an increase in the expression of epidermal growth factor receptor (EGFR). Epidermal growth factor (EGF) also stimulated terminal branch formation in the developing lung. Unlike RA, EGF stimulated proliferation in both epithelial cells and mesenchymal cells in monoculture. In comparison, transforming growth factor alpha, which also binds to the EGFR, elicited no response. We conclude that RA stimulates cell proliferation and branching activity in the developing mouse lung by a mechanism involving epithelial-mesenchymal interactions. The effect is, in part, produced by stimulation of EGFR expression, with the resulting amplification of the cellular response to EGF or other EGFR ligands. In this process the mesenchyme provides a paracrine support to the epithelium, otherwise unresponsive to RA. Further studies identified the mesenchyme as a major source of EGF in the embryonic lung, suggesting that mesenchymal EGF may represent a paracrine factor involved in the epithelial response to RA. PMID- 8405672 TI - Evidence for the involvement of internal calcium stores during serotonin-induced meiosis reinitation in oocytes of the bivalve mollusc Ruditapes philippinarum. AB - In contrast to the situation found in the bivalves Barnea candida and Spisula solidissima, prophase-arrested oocytes of Ruditapes philippinarum cannot be fertilized when removed from the ovary. They must first undergo germinal vesicle breakdown under the influence of the neurohormone serotonin (5-HT), which drives them to a second block occurring in metaphase of the first maturation division. In the studies described in this paper, we investigate the possibility that calcium is involved as a second messenger in controlling this first step in the reinitiation of meiosis. Our data show that, in addition to 5-HT, ionophore, thapsigargin, and the weak bases ammonia and procaine can also induce prophase arrested oocytes of Ruditapes to resume meiosis. 5-HT, thapsigargin, and ammonia all trigger a surge of intracellular Ca2+ and are effective even in the absence of external Ca2+. That such Ca2+ transients, which are enhanced in the presence of external Ca2+, actually play a key role in the process of meiosis reinitation is shown by the fact that loading the oocytes with BAPTA/AM or treating them with D-600 blocks maturation. In contrast, excess KCl, which has been shown to trigger meiosis reinitiation of prophase-arrested oocytes of Barnea and Spisula and to activate metaphase I-arrested oocytes of Ruditapes, does not produce any significant intracellular Ca2+ transient nor does it reinitiate meiosis, when added to Ruditapes prophase-arrested oocytes. These data suggest that such voltage-operated Ca2+ channels may only appear during the course of maturation and that both intracellular and extracellular Ca2+ are involved in triggering 5 HT-dependent release from the prophase block in this species. PMID- 8405673 TI - Expression of the helix-loop-helix factor Id during mouse embryonic development. AB - Expression and regulation of the helix-loop-helix (HLH) protein Id in cardiac cells and its negative effect on cardiac gene expression indicate that Id may play a role in cardiac development. To investigate this issue further, we have performed in situ analysis of Id mRNA in developing mouse embryos from Embryonic Days 7 to 15.5 and in neonatal heart. Consistent with its postulated role as an inhibitor of differentiation, Id mRNA was expressed in specific populations of proliferating mesenchymal and epithelial tissues, with its expression declining as differentiation progressed. In addition, patterns of Id mRNA hybridization overlapped considerably with those of two other developmentally important genes, twist and hox 7/7.1, of the HLH and homeobox family, respectively. Id mRNA hybridization was not observed in early epithelial somites or in the myotomal compartment of the somite, suggesting that Id is unlikely to play an inhibitory role in these early myogenic precursors. Id mRNA was expressed in specific regions of the developing nervous system. In the heart, Id mRNA was expressed at high levels in developing cardiac cushions and ridges of the outflow tract and continued to be expressed at high levels in valvular tissue of the neonatal heart. PMID- 8405674 TI - Developmental regulation of the Drosophila tropomyosin II gene in different muscles is controlled by muscle-type-specific intron enhancer elements and distal and proximal promoter control elements. AB - Transcriptional control of the Drosophila tropomyosin II gene muscle promoter has been investigated by expressing TmII promoter lacZ reporter gene constructs in P element-mediated transformed flies. A TmII/lacZ reporter gene containing 243 bp of upstream sequence, the first exon, the first intron, and 72 bp of the second exon was expressed in all muscles of embryos, larvae, and adults. Deletion of upstream sequences between -243 and -22 bp only reduced the levels of transgene expression in muscle while deletion of the intron eliminated expression. Analysis of deletions within the first intron indicated that a 454-bp muscle enhancer region, from +167 to +621, was required for high levels of transgene expression in all larval and adult muscles. When this region was deleted low levels of expression still occurred in larval and adult somatic and visceral muscles; however, there was no detectable expression in adult indirect flight and jump muscles. The 454-bp muscle enhancer region was also able to drive muscle-specific expression when placed upstream of a heterologous hsp70 promoter; however, three subfragments of the 454-bp region were unable to drive expression of the hsp70 promoter, suggesting that this region may contain multiple interacting cis-acting elements. Interestingly, the 454-bp region was inactive when placed upstream of a TmII promoter construct containing upstream DNA and most of the first exon but was active when additional exon and intron DNA was included, indicating that additional promoter elements are located in this region. Thus TmII transcription is controlled by multiple muscle type-specific cis-acting control elements and upstream and downstream promoter control elements. PMID- 8405675 TI - Coordinate regulation of Drosophila tropomyosin gene expression is controlled by multiple muscle-type-specific positive and negative enhancer elements. AB - Muscle development involves the coordinated regulation of transcription of muscle type-specific genes and their encoded proteins during myogenesis. We show here that transcriptional regulation of the Drosophila tropomyosin I (TmI) gene during myogenesis is under the control of at least two muscle enhancer regions located within the first intron of the gene. Together these enhancer regions contain multiple muscle-type-specific positive and negative cis-acting elements which together contribute toward full expression of the gene. One of these enhancers is contained within a 355-bp fragment that is sufficient to direct high levels of temporally regulated expression from a heterologous promoter in all muscles of transgenic flies. Dissection of this enhancer region into smaller fragments has allowed us to identify a 91-bp enhancer fragment sufficient for directing expression in all somatic and visceral muscles of the larva and adult but not in the indirect flight muscles and tergal depressor of the trochanter or jump muscles of the adult. We also show that this somatic/visceral muscle element(s) can be repressed through an adjacent negative control region, suggesting that the regulation of expression in these muscles is under dual control during both phases of myogenesis. We propose a model in which transcriptional regulation of the Drosophila TmI gene is controlled by the cooperative interaction of multiple positive and negative cis-acting regulatory elements that control the temporal and muscle-type pattern of expression. The distribution of enhancer elements and their control of TmI gene expression are similar to those regulating transcription of the muscle promoter of the TmII gene and provide a framework for the coordinate expression of the two genes. PMID- 8405676 TI - Type X collagen degradation in long-term serum-free culture of the embryonic chick tibia following production of active collagenase and gelatinase. AB - Type X collagen has a very limited distribution during skeletal development in regions of hypertrophic cartilage destined for degradation. In solution assay, type X collagen is degraded to a 32-kDa cleavage product which is resistant to further degradation, suggesting this product may have a function in skeletal development. In this study, we have identified the 32-kDa cleavage product of type X collagen present in the conditioned media (CM) during incubation of isolated 12-day chick tibiae in the absence of serum. In this culture system, chondrocytes throughout the tibial cartilages hypertrophied and deposited type X collagen within their matrix. During culture, the cartilage matrix was degraded in two stages. First proteoglycan was lost followed by degradation of the collagenous components. Collagen degradation was accompanied by the release of active interstitial collagenase and gelatinase into the CM. Purified type X collagen incubated in this CM was cleaved to form a 32-kDa product which was resistant to further degradation. This cleavage product has the same electrophoretic mobility as the 32-kDa chain produced by purified human collagenase. PMID- 8405677 TI - Modulation of HGF-induced tubulogenesis and branching by multiple phosphorylation mechanisms. AB - MDCK cells cultured in Type I collagen gels can be induced to develop branching tubular structures with demonstrable lumens in the presence of hepatocyte growth factor (HGF). As we have now shown by immunofluorescent localization of specific marker proteins, these tubules retain apical-basolateral polarity. However, the secondary signaling events which lead to these characteristic morphogenetic changes induced by HGF remain largely unelucidated. In order to examine these signaling events, particularly the role of protein phosphorylation in the formation of branching tubular structures, Madin-Darby canine kidney (MDCK) cells in collagen gels were treated with HGF plus well-characterized agents that affect protein phosphorylation. We quantified the formation of branching processes, an early step in the development of tubular structures in this model. Protein kinase C (PKC) inhibition resulted in more complex branching processes in the presence of HGF, when compared with HGF alone. In contrast, treatment with activators of protein kinase A (PKA), as well as calmodulin antagonists, caused a marked decline in process formation. Consistent with an important role for protein phosphorylation in HGF-induced morphogenesis, protein phosphatase inhibition by okadaic acid or calyculin A was found to markedly inhibit process formation. Tyrosine kinase (TK) inhibition also decreased the percentage of processes. This is consistent with data indicating that one of the HGF receptors is identical to the c-met protooncogene product, which is known to possess TK activity. Our results suggest that the HGF-mediated induction of branching processes in MDCK cells, an early step in the development of branching tubular structures, can be modulated by multiple phosphorylation mechanisms including those mediated by PKC, PKA, and Ca2+/calmodulin-dependent kinase(s). We discuss how these phosphorylation events may play crucial roles in determining the degree of tubule formation and their length, as well as the extent of their arborization during the early development of epithelial tissues. PMID- 8405678 TI - Descriptive and experimental analysis of the epithelial remodellings that control semicircular canal formation in the developing mouse inner ear. AB - The inner ear of the mouse develops from a roughly spherical epithelial vesicle, the otocyst, which undergoes a series of complex shape changes to produce the functionally important parts of an adult inner ear; in particular, a coiled cochlea--which houses the auditory apparatus, a saccule and utricle containing sensors of gravity and linear acceleration, and three precisely shaped and oriented semicircular canals, with which angular acceleration is detected. This paper follows the development of the shape of the mouse inner ear from simple otocyst until a stage when the vesicle has become a rather squat miniature model of its adult self. We have been able to visualize clearly these complex shape changes by injecting an opaque marker into the lumina of a series of fixed ears. We have further concentrated on the mechanism of formation of the semicircular canals using light-, electron-microscopic, and dye-marking techniques. Classic embryological texts describe the canals developing from outpocketings of the epithelial ear rudiment, whose opposite walls meet, fuse, and "disappear" in the central canal plate region to leave a tube of epithelium encircling their margin. To trace the fate of these "disappearing" epithelial cells we have dye-marked all of the otic epithelial cells at a stage prior to canal formation in mouse embryos which we then grow in roller culture until their canals have formed. From these marking experiments we show that the disappearing cells of the canal plate neither die nor become transformed into mesenchymal cells, but rather, most are retracted back into the canal tube epithelium at either side of the site of fusion. We speculate that this mechanism of epithelial resorption may be a common way of "losing" epithelial cells during embryonic morphogenetic remodellings. PMID- 8405679 TI - Inhibition of migration of neural crest-derived cells by the abnormal mesenchyme of the presumptive aganglionic bowel of ls/ls mice: analysis with aggregation and interspecies chimeras. AB - The terminal bowel is congenitally aganglionic in ls/ls mice. The condition has been associated with an overabundance of laminin and other matrix molecules. Aggregation ls/ls<==>C3H chimeric mice and interspecies mouse<==>quail chimeras were constructed to test the hypothesis that the aganglionosis arises because the ls/ls gut and not the neural crest is abnormal. Demonstration of beta glucuronidase activity permitted genotypically ls/ls and C3H cells to be distinguished in the ls/ls<==>C3H chimeras. Aganglionosis did not occur in the ls/ls<==>C3H mice and ls/ls neurons were observed in the terminal bowel. Following bactransplantation of control segments of mouse gut into quail host embryos, mouse cells migrated to host targets normally colonized by cells from the neural crest; moreover, quail crest-derived cells entered the mouse gut. In contrast, cells did not migrate to these targets from presumptive aganglionic ls/ls bowel and quail crest-derived cells neither entered the ls/ls gut nor migrated through it. Laminin immunoreactivity was present in the backgrafts of murine colon and was far more abundant and widespread in those from ls/ls than in those from control animals. These data suggest that the presumptive aganglionic ls/ls bowel does not contain crest-derived cells because these cells, which are normal in ls/ls mice, do not enter it. This failure of colonization may be related to the premature formation of neurons outside the abnormal gut, a response that may be promoted by the excessive secretion of laminin by the ls/ls enteric mesenchyme. PMID- 8405680 TI - Expression of activin transcripts in follicle cells and oocytes of Xenopus laevis. AB - Activin is a potent inducer of axial mesoderm in vitro and may have a similar role in vivo. Xenopus laevis eggs contain significant amounts of activin or activin-like factors, but maternal activin transcripts have not been detected in Xenopus eggs. The maternal activin protein might be translated from activin beta A or beta B mRNAs that are transiently expressed during oogenesis, or activin polypeptides might be transferred from follicle cells to oocytes. To assess these possibilities we studied activin mRNAs in follicle cells and oocytes by reverse transcriptase-polymerase chain reaction (RT-PCR) and RNA blotting. Activin beta A, beta B1, and beta B2 transcripts occur in follicle cells; among them, beta A mRNA is by far the most abundant. Activin beta A and beta B1 mRNAs were not detected by RT-PCR in the corresponding stage IV oocytes, but activin beta B2 transcripts were found at low levels. These observations are consistent with synthesis of activin beta A and possibly beta B polypeptides in follicle cells followed by their secretion and uptake by oocytes, although synthesis of activin beta B2 in the oocytes could make a contribution to the maternal activin pool. PMID- 8405681 TI - Sea urchin egg 100-kDa dynamin-related protein: identification of and localization to intracellular vesicles. AB - We have initiated a study to identify possible tubulin dimer binding proteins from unfertilized sea urchin eggs utilizing tubulin dimer coupled to Sepharose 4B. A 100-kDa polypeptide has been isolated and its interaction with tubulin/microtubules examined. Solution assembly studies indicated the 100-kDa polypeptide potentiated microtubule assembly. The 100-kDa protein also bundled and cosedimented with polymerized microtubules. Rabbit polyclonal anti-egg 100 kDa antibodies cross-reacted with a 100-kDa polypeptide in the eggs of several different sea urchin species and with mammalian brain dynamin, a 100-kDa microtubule binding protein. These biochemical characterizations and immunological results suggest that the sea urchin 100-kDa microtubule binding protein appears to be related to the dynamin protein family. To gain insight into the functional role of sea urchin egg 100-kDa dynamin-related protein, we immunolocalized egg 100-kDa protein in cryosections and isolated cortices of unfertilized eggs. Egg 100-kDa protein was localized to the monolayer of cortical granules that underlie the plasma membrane and to the inner cytoplasm. Immunolocalizations of egg 100-kDa protein in cortices isolated from fertilized eggs detailed the redistribution of the egg 100-kDa onto cortically positioned vesicles. Egg 100-kDa dynamin-related protein was expressed throughout early development with approximately equivalent amounts present in the unfertilized egg and embryo through early gastrulation, followed by an abrupt decrease in the level of expression by the pluteus larval stage. PMID- 8405682 TI - Glial cells of the O-2A lineage bind preferentially to N-cadherin and develop distinct morphologies. AB - Environmental cues guiding glial cell behavior during development or regeneration of the nervous system are provided by both soluble and nondiffusible factors. We examined the influences of purified extracellular matrix molecules and cell adhesion molecules on the development and proliferation of glial cells from neonatal rat optic nerves. Dissociated optic nerve glia were plated on fibronectin, laminin, collagen type IV, L1, N-cadherin, and N-CAM. Cultures were grown in chemically defined medium to promote formation of oligodendrocytes. Other cultures were grown in 10% serum to support type-1 astrocytes and the differentiation of O-2A progenitor cells to type-2 astrocytes. In short-term adhesion assays to measure cell affinity for the different substrates, cells in the O-2A lineage bound preferentially to N-cadherin while type-1 astrocytes preferentially bound to extracellular matrix components. The cells in the O-2A lineage also developed distinctive morphologies on different substrates after incubation for 4 days. Type-2 astrocytes and oligodendrocytes produced very large membranous expansions on N-cadherin. Measurements of BrdU incorporation indicated that the substrates did not significantly influence cell proliferation rates. Our results showed that O-2A progenitor cells, oligodendrocytes, type-1 astrocytes, and type-2 astrocytes possess different complements of receptors for the adhesion molecules in their environment and that their morphological differentiation can be dramatically altered by these extracellular signals. PMID- 8405683 TI - Nuclear and cytoplasmic pH increase at fertilization in Urechis caupo. AB - Intracellular pH (pHi) was measured in Urechis caupo (Echiura) eggs during fertilization using the pH-sensitive dye BCECF [bis(carboxyethyl)carboxyfluorescein] and fluorescence microscopy. When eggs were inseminated at pH 8, pHi began to rise 22-36 sec (n = 7) after sperm contact and reached a plateau by 3 min (2.8 +/- 1 SD; n = 14). The net increased was 0.25-0.3 pH units and the alkalinization persisted through 1 hr after insemination (after second polar body formation). Separate measurements of germinal vesicles and cytoplasm revealed that pH rose dramatically within the nuclei well before germinal vesicle breakdown (GVBD), as well as in cytoplasm. When eggs were fertilized at pH 6.9 (previously shown to inhibit proton release and egg activation despite sperm entry; see Paul, 1970, 1975; Holland et al., 1984), there was no net pH increase in either cytoplasm or germinal vesicles. When fertilization was at pH 7.45, the average pHi increase in whole eggs was 58% of that at pH 8, and 61% of the eggs activated partially or completely. These results show that the pHi rise is correlated with GVBD and egg activation. PMID- 8405684 TI - Glial cell mitogens bFGF and PDGF differentially regulate development of O4+GalC- oligodendrocyte progenitors. AB - The oligodendrocyte lineage in cerebrum is characterized by the expression of immunologically identified surface antigens resulting in the sequential appearance of three distinct phenotypes, A2B5+O4-, O4+GalC-, and O4+GalC+. In the present study we have placed O4+GalC- progenitors immunopanned from premyelinating rat cerebrum into a basal, defined medium that by itself does not support well either their proliferation or survival. The response of these progenitor cells to platelet-derived growth factor (PDGF) and basic fibroblast growth factor (bFGF) was then examined. The results demonstrate that both PDGF and bFGF stimulated proliferation and short-term survival of newly cultured cells, but that their effect on the course of O4+GalC- differentiation was strikingly different. PDGF delayed postmitotic development by transiently reverting (ED50 = 3 ng/ml) O4+GalC- progenitors to A2B5+O4- preprogenitor-like cells that subsequently differentiated even in the continued presence of PDGF. bFGF restored mitogenic activity of the O4+GalC- progenitors to a saturable level at low doses (ED50 = 1 ng/ml); doses of bFGF > or = 10 ng/ml impaired differentiation of the progenitors into GalC+ cells and were also mitogenic for newly differentiated GalC+ oligodendrocytes. These data imply that bFGF supplants PDGF as a mitogen during lineage progression from A2B5+O4- to O4+GalC- progenitors. Lineage reversion of O4+GalC- cells in response to PDGF is suggested as a mechanism for facilitating remyelination by triggering the proliferative expansion of O4+GalC- progenitor-like cells persisting into adulthood. PMID- 8405685 TI - Hybrid sterility-6: a mouse t complex locus controlling sperm flagellar assembly and movement. AB - Mouse t haplotypes alter sperm differentiation, resulting in abnormal sperm movement and sterility. In previous studies, a locus responsible for hybrid sterility in the genus Mus,Hst-4, was mapped to the distal inversion of the t complex on chromosome 17. Here we report the identification and characterization of two additional hybrid sterility loci, Hst-5 and Hst-6, that map to the same inversion. We further show that an abnormality in sperm flagellar curvature deriving from interactions between t haplotypes and the M. spretus allele of Hst 6 is indistinguishable from one exhibited by sperm from mice carrying two t haplotypes. Additionally, we demonstrate that this latter phenotype maps to the distal inversion of t haplotypes. Morphological and functional studies of Hst-6 mutant sperm also imply that the product(s) of Hst-6 is a spermatogenic-specific protein, important for assembly and function of the sperm axoneme. Thus, Hst-6 provides direct access to the molecular basis of t haplotype-specific alterations in sperm function that emanate from the t complex distal inversion. PMID- 8405686 TI - Biochemical differentiation and intercellular interactions of migratory gonadotropin-releasing hormone (GnRH) cells in the mouse. AB - GnRH cells are first detected in the olfactory placode of the mouse on Gestational Day 11.5 (E11.5). Between E12.5 and E15.5 they migrate across the nasal septum and by E16.5 attain their adult distribution within the forebrain. In the present study, we used immunocytochemistry at the light and electron microscopic level to study the biochemical and morphological differentiation of gonadotropin-releasing hormone (GnRH) neurons and the cellular associations that they make during this migratory process. On E12.5, when the majority of GnRH cells are still in the nasal septum, only 15% of the population can process the pro-GnRH precursor to the amidated decapeptide. Two days later (E14.5), when most of the cells have advanced into the forebrain, 79% contain mature GnRH. In keeping with these light microscopic observations, ultrastructural analysis reveals that on E12.5 GnRH immunologic reaction product is confined to the outer nuclear envelope and rough endoplasmic reticulum. By E14.5 these migratory cells in the nasal septum have more reaction product in the rough endoplasmic reticulum and some of the Golgi cisternae are also immunopositive. Neurosecretory granules, some of which are immunoreactive, also appear at this stage. We had anticipated that the expression of GAP-43 would coincide with axonal elongation and pathfinding in GnRH neurons. Instead, GAP-43 was expressed at the early migratory stage of GnRH cells and its expression declined rapidly after these neurons had entered the forebrain and commenced axonal outgrowth. Hence, on E12.5, 62.3% of GnRH cells in the nasal septum are immunopositive for GAP-43, while only 12.6% of the forebrain population at the same embryonic stages express the protein. Similarly, on E14.5 GAP-43 is expressed in 50.7% of the nasal septum GnRH cells but only 11.6% of cells in the E14.5 forebrain express this protein. While in the nasal septum, GnRH neurons migrate only within the confines of the olfactory and vomeronasal axonal fascicles. During this part of the migratory route, the cells maintain close membrane apposition with each other and with the axons and ensheathing glia of the nerve fascicles. Once in the forebrain, GnRH neurons no longer maintain these associations, nor do they follow any defined anatomical structure. These findings indicate that although GnRH cells express their unique neuropeptide early in their ontogeny, their differentiation continues and is coordinated with their migration.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8405687 TI - Three populations of migrating amphibian embryonic cells utilize different guidance cues. AB - Previous investigations designed to identify the molecule(s) governing the directed migration of the amphibian pronephric duct (PND) revealed a requirement for a glycosyl phosphatidylinositol (GPI)-linked cell surface molecule, possibly the ectoenzyme alkaline phosphatase (AP). Cranial neural crest cells (CNC) grafted to the flank migrate along the same pathways as the PND, suggesting that PND and CNC guidance systems might have a common molecular basis. Both PND and CNC migration pathways display AP. The present experiments demonstrate, however, that GPI-linked molecules on these pathways are not required for migration of either cell type. Because PND cells themselves express AP prominently but CNC cells do not, we asked whether the GPI-linked molecule required for PND migration resides on the PND cells themselves. Treatment of PND cells with phosphatidylinositol-specific phospholipase C, an enzyme that removes GPI-linked proteins, prevents their migration. Transplantation experiments show that although CNC cells are capable of following PND migration pathways, the converse is not the case. Extension of the transplantation experiments to include trunk neural crest (TNC) cells indicates that although CNC cells can follow PND guidance information on the flank, PND, CNC, and TNC cells all normally utilize different molecular cues to guide their migrations in situ. PMID- 8405688 TI - Complete mouse egg activation in the absence of sperm by stimulation of an exogenous G protein-coupled receptor. AB - Sperm-induced egg activation has several features in common with signal transduction pathways mediated by ligand-receptor-effector systems. G proteins are postulated to act as signal transducers regulating the plethora of cellular responses constituting egg activation that occur in response to sperm. We report that acetylcholine treatment of mouse eggs expressing the human m1 muscarinic receptor, a G protein-coupled receptor, results in the full complement of events of egg activation, including ZP2 to ZP2f conversion, recruitment of maternal mRNAs, pronuclear formation with subsequent DNA synthesis, cleavage to the two cell stage, and activation of the embryonic genome. The induction of a full complement of sperm-induced egg activation events by ACh in the absence of sperm strongly suggests that sperm-induced egg activation is mediated by egg G proteins. PMID- 8405689 TI - Evidence suggesting involvement of a unique human sperm steroid receptor/Cl- channel complex in the progesterone-initiated acrosome reaction. AB - Progesterone (4-pregnen-3-20-dione) initiates the acrosome reaction (AR) of human sperm in vitro. Anesthetic progestins (e.g., progesterone) potentiate gamma aminobutyric acid (GABA) activation of neuronal GABAA receptor/Cl- channel complexes (GBRCs), and some can activate neuronal GBRCs in the absence of GABA. Here, we study whether human sperm acrosome reaction initiation by progesterone and several other progestins involves steroid interaction with a sperm GBRC. Anesthetically active 3 alpha-OH isomers of two progesterone metabolites, as well as the synthetic 3 alpha-OH anesthetic steroid Alphaxalone, their 3 beta-OH isomers (inactive as anesthetics), or progesterone, were added to capacitated human sperm. The 3 alpha-OH isomers initiated the AR, and the 3 beta-OH isomers were without effect. Moreover, the 3 alpha-OH isomers were less effective than progesterone, a progestin that does not activate neuronal GBRCs in the absence of GABA. Also, alphaxalone, a very potent activator of neuronal GBRCs, was the least active of the progestins tested. GABA was not detected in human sperm. Preincubation of capacitated sperm with GBRC Cl- channel blockers dramatically reduced the progesterone-initiated AR, and sperm were unable to undergo the progesterone-initiated AR in Cl(-)-deficient medium. AR initiation by the calcium ionophore ionomycin was not inhibited in Cl(-)-deficient medium or by GBRC Cl- channel blockers. Indirect immunofluorescence, using a monoclonal antibody to the bovine cerebral cortex GABAA receptor alpha-subunit, localized immunoreactivity in live and fixed sperm as a fluorescent band in the sperm plasma membrane overlying or near the narrow equatorial segment region of the acrosome. Immunoblotting using this antibody detected two major bands with apparent molecular weights of 50 kDa (as reported in other cell types) and 75 kDa (not reported in other cells). Our data suggest that (1) progesterone exerts its effect on capacitated human sperm at least partially by interaction with a unique sperm steroid receptor/Cl- channel complex resembling a neuronal GBRC but apparently possessing a different sensitivity to progestins; and that (2) this interaction results in an increased Cl- flux essential to AR initiation. PMID- 8405690 TI - Primitive-streak origin of the cardiovascular system in avian embryos. AB - The origin of the avian cardiovascular system from the primitive streak has been mapped by the construction of quail/chick transplantation chimeras, the use of QH 1 (an antiquail endothelial/endocardial cell marker), and injections of a vital fluorescent dye (DiI) into the primitive streak. Our studies reveal that the prospective heart, including its endocardial and myocardial layers and the adjacent parietal pericardium, occupies much of the rostral half of the primitive streak at early primitive-streak stages of gastrulation. The heart originates from the primitive streak in roughly rostrocaudal sequence (i.e., the prospective bulbus cordis arises more rostrally in the streak than does the prospective ventricle, which in turn arises more rostrally than does the prospective sinus venosus), and all layers of the heart at each of its rostrocaudal subdivisions originate in concert from the same level of the primitive streak. Moreover, all rostrocaudal levels of the primitive streak at gastrula and neurula stages contain prospective endothelial cells. Again, these cells are rostrocaudally ordered within the streak, such that head blood vessels (both arteries and veins) arise at more rostral streak levels than do trunk blood vessels. Ingression of cardiogenic cells is complete by midprimitive-streak stages, and the position formerly occupied by prospective cardiogenic cells within the streak becomes occupied by ingressing prospective somitic cells. Additional studies on the state of commitment of prospective cardiogenic and endothelial cells during their ingression through the primitive streak are underway. PMID- 8405691 TI - Multiple controls over the efficiency of translation of the mRNAs encoding transition proteins, protamines, and the mitochondrial capsule selenoprotein in late spermatids in mice. AB - The mRNAs encoding protamines 1 and 2, transition proteins 1 and 2, and the mitochondrial capsule selenoprotein are translationally repressed with long poly(A) tracts in early spermatids and translationally active with heterogenous shortened poly(A) tracts in late spermatids (Kleene, 1989). In the present study, the spacing of ribosomes on the translationally active forms of each mRNA was calculated from the length of the coding region and the polysome size determined by sucrose gradient and Northern blot analysis. In addition, the rate of initiation of these five mRNAs was compared in the reticulocyte cell-free translation lysate. Our results reveal at least four additional forms of translational control over these mRNAs: (1) The vast majority of the active forms of the transition protein 1 mRNA and both protamine mRNAs sediment with polysomes in which the ribosomes are spaced closer than is typical of mammalian mRNAs (31 38 vs 80-100 bases apart). This implies that the rate of initiation is unusually fast and/or that the rate of elongation is unusually slow. (2) The mitochondrial capsule selenoprotein mRNA also initiates efficiently in vivo and in vitro, but sediments with polysomes in which the ribosomes are spaced wider than on the protamine mRNAs. The small size of these polysomes can be explained by inefficient insertion of selenocysteine residues at UGA codons. (3) The transition protein 2 mRNA is translated on small polysomes and a relatively large fraction sediments as free mRNPs in vivo. In addition, the transition protein 2 mRNA initiates translation inefficiently at high mRNA concentration and efficiently at low mRNA concentration in vitro. These observations suggest that the transition protein 2 mRNA may be translated inefficiently because it is a weak competitor for a limiting initiation factor. (4) Since low levels of cycloheximide fail to increase the polysome loading of transition protein 2 mRNA in culture, active single ribosomes may also be limited in late spermatids. PMID- 8405692 TI - Diacylglycerol content of Chaetopterus oocytes during maturation and fertilization. AB - The resumption of meiotic maturation in Chaetopterus oocytes is believed to be initiated by activation of protein kinase C (pkC). Since the physiological activator of pkC is diacylglycerol (DG), we examined the DG content of the oocytes before and during germinal vesicle breakdown (GVBD). Unstimulated oocytes contained 366 +/- 34 fmol DG per cell. This value increased to approximately 430 500 fmol 3-5 min after normal induction of GVBD and declined thereafter to 147 +/ 7 fmol per cell. Fertilization increased the DG levels slightly, 3-5 min after insemination. These results strongly support the hypothesis that pkC is a physiological activator of GVBD in this species. PMID- 8405693 TI - Lessons from the Diabetes Control and Complications Trial. PMID- 8405694 TI - Implications of the Diabetes Control and Complications Trial. American Diabetes Association. PMID- 8405695 TI - Metabolic consequences of sustained suppression of free fatty acids by acipimox in patients with NIDDM. AB - To examine whether overnight suppression of free fatty acid levels reduces hepatic glucose production, 20 NIDDM patients were given a slow-release formulation of the antilipolytic agent acipimox, in a double-blind crossover manner at bedtime for 4 wk. During acipimox treatment, serum free fatty acid concentrations were suppressed between 2400 and 0600 by 64% (P < 0.001), but no reduction in hepatic glucose production was observed (2.16 +/- 0.16 vs. 2.23 +/- 0.16 mg.kg-1 x min-1, acipimox vs. placebo). In contrast, from 0800 to 2000 a sustained 50% rise occurred in serum free fatty acids (P < 0.001). As a consequence, the 24-h area under the free fatty acid curve was similar during both treatment periods. In the morning, the rise in free fatty acid concentration occurred despite identical serum acipimox concentrations as those measured at midnight, when free fatty acid levels were suppressed. Although energy expenditure was higher (P < 0.05) during periods of elevated free fatty acid levels, the sums of energy expenditure measured in the morning and in the evening were similar during the acipimox and placebo periods. To exclude that the free fatty acid rise was caused by administration of acipimox only once at bedtime, additional experiments were performed administering acipimox every 2 h for 4 days. Despite similar acipimox concentration on day 1 and day 4 of this frequent dosing regimen, the free fatty acid concentrations were significantly higher on day 4 compared with day 1 (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405696 TI - Postprandial free fatty acid kinetics are abnormal in upper body obesity. AB - Excess free fatty acid release (rate of appearance) is seen in overnight postabsorptive, upper body obese women and, if present postprandially, could contribute to glucose intolerance. These studies examine the antilipolytic effect of a mixed meal in upper body obese, lower body obese, and nonobese women and the contribution of meal triglyceride fatty acids to circulating free fatty acids. Eight upper body obese, 8 lower body obese, and 8 nonobese age-matched, premenopausal women were studied. Systemic oleate Ra ([3H]oleate) was measured before and after an evening meal that contained triolein labeled with [14C]triolein as the only source of fat. Premeal oleate Ra was greater in both upper body obese and lower body obese women than nonobese women. The nadir of total oleate Ra occurred 90-240 min postprandially and was less (P < 0.01) in nonobese and lower body obese women (63 +/- 10 and 87 +/- 17 mumol/min) than in upper body obese women (140 +/- 21 mumol/min). Meal oleate Ra contributed substantially to total oleate Ra. The nadir for endogenous oleate Ra in nonobese and lower body obese women was less (P < 0.01) than that observed in upper body obese women. We conclude that the antilipolytic effect of a mixed meal is reduced in upper body obese women and that meal triglyceride fatty acids contribute significantly to postprandial free fatty acid flux. PMID- 8405697 TI - Complement lytic activity has no role in the pathogenesis of autoimmune diabetes in NOD mice. AB - The NOD mouse is widely used as a model of organ-specific autoimmunity because it develops specific autoimmune destruction of pancreatic beta-cells. Although it is clear that T-cells and monocytes are necessary for beta-cell destruction, humoral factors, such as antibodies and complement, may also contribute to tissue damage. Attempts to cure diabetes in experimental models by immunoisolation of transplanted islets has raised the need to protect the islets from the relatively small components of the complement cascade. In this study, we report that NOD mice have no complement lytic activity and that the exclusion of complement is unnecessary in this model. Sera from young NOD mice were unable to lyse sheep red blood cells coated with rabbit antibody. Lytic activity of NOD sera was reconstituted by mixing with C4-deficient CBA sera, but not C5-deficient DBA/2 sera, indicating the presence of C4, but the absence of C5 activity in NOD sera. Lytic activity of NOD sera could be reconstituted with human C5 electrofocused in polyacrylamide gel. The polymerase chain reaction was used to amplify fragments from genomic DNA corresponding to the region of Hc (the gene encoding C5) in DBA/2 mice, which carries a 2-base pair deletion responsible for the lack of C5 protein expression in these mice. DBA/2 and NOD mice from several colonies produced a fragment 2 bases shorter than that generated from the wild-type allele in BALB/c mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405698 TI - Dietary carbohydrate enhances intestinal sugar transport in diabetic mice. AB - The rate of intestinal absorption of sugars and their site of absorption determine postprandial plasma glucose concentrations. Does chronic consumption of high-carbohydrate, high-fiber, low-fat diets of the type recommended by many diabetes associations induce adaptive changes in transport and metabolism of sugars in the small intestine? Control and STZ-induced diabetic (> 60 days diabetic) mice were fed high-carbohydrate or no-carbohydrate rations for 7 days. Brush-border glucose and fructose uptake per milligram increased 2 times with dietary carbohydrate in both diabetic and control mice; uptake, however, did not differ between diabetic and control mice. Compared with the distal small intestine, glucose uptake per milligram was 2 to 6 times higher in the proximal and middle regions, and enhancement of uptake by diet was limited to these regions. Changes in site density of intestinal glucose transporters as determined by specific phlorizin binding were tightly correlated with changes in brush border glucose uptake per milligram. There were neither diabetes- nor diet induced changes in the Kd of specific phlorizin binding, in the amount of glucose absorbed per transporting site, or in passive glucose permeability. Intestinal weights, wt/cm, intestinal length, and mucosal mass increased significantly with diabetes, and sugar transport per centimeter and per small intestine was up to 60% greater in diabetic mice. Dietary carbohydrate stimulated specific sucrase activity in the proximal small intestine of both diabetic and control mice. Chronic diabetes enhances sugar transport by nonspecific increases in intestinal mass.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405699 TI - Effects of insulin on fatty acid reesterification in healthy subjects. AB - The effects of insulin on triacylglycerol/fatty acid cycling (fatty acid reesterification) were studied in 12 normal subjects during euglycemic hyperinsulinemia with the use of stable isotope dilution analysis ([2H5]glycerol and [1-13C]palmitate) in combination with indirect calorimetry. During basal conditions, 5.6 +/- 0.6 mumol.kg-1 x min-1 of fatty acid were released of which approximately 3.3 mumol.kg-1 x min-1 were oxidized and approximately 2.2 mumol.kg 1 x min-1 were reesterified. A minority of the recycled fatty acid, (0.8 +/- 0.4 mumol.kg-1 x min-1) never left the intracellular space before being reesterified (intracellular triacylglycerol/fatty acid cycling), whereas the majority (1.2 +/- 0.4 mumol.kg-1 x min-1) were first released into the extracellular space and then reesterified in various organs (extracellular triacylglycerol/fatty acid cycling). In response to insulin, fatty acid release declined by 71% (from 5.6 +/ 0.6 to 1.6 +/- 0.2 mumol.kg-1 x min-1). Fatty acid oxidation (measured by indirect calorimetry) declined by 55% (from 3.3 +/- 0.3 to 1.5 +/- 0.3 mumol.kg-1 x min-1) and total triacylglycerol/fatty acid cycling was completely suppressed (from 2.2 to 0.0 mumol.kg-1 x min-1). Fatty acid release, oxidation, total and extracellular triacylglycerol/fatty acid cycling all correlated positively with plasma fatty acid concentrations. These data showed that insulin profoundly suppressed fatty acid release, oxidation as well as reesterification of those fatty acids that had entered the extracellular compartment. They suggested that physiological concentrations of insulin suppressed extracellular fatty acid reesterification primarily by inhibiting lipolysis. PMID- 8405700 TI - Effects of high glucose concentrations and epalrestat on sorbitol and myo inositol metabolism in cultured rabbit aortic smooth muscle cells. AB - To clarify the relationship between abnormality of sorbitol and/or myo-inositol metabolism caused by hyperglycemia and diabetic macroangiopathy, we investigated the effects of high glucose concentrations and epalrestat, an aldose reductase inhibitor, on the metabolism of sorbitol and myo-inositol in cultured rabbit aortic smooth muscle cells. In cells incubated in the presence of 30 mM glucose for 72 h, the sorbitol content increased approximately 4.5-fold, and the myo inositol level decreased by 55% compared with control values. Kinetic analysis of high-affinity myo-inositol uptake suggested that smooth muscle cells exposed to high glucose concentrations exhibited a noncompetitive type of inhibition characterized by ouabain-sensitive, energy-dependent active transport. Epalrestat blocked glucose-induced changes in sorbitol and myo-inositol metabolism, suggesting that these changes were caused by the accumulation of sorbitol in the cells. These metabolic changes may impair function of smooth muscle cells, contributing to the pathology of diabetic atherosclerosis, especially Monckeberg's calcific medial sclerosis. The use of an aldose reductase inhibitor may prevent these glucose-induced changes. PMID- 8405701 TI - Tolerance to IDDM induced by CD4 antibodies in nonobese diabetic mice is reversed by cyclophosphamide. AB - IDDM can be induced in nonobese diabetic (NOD) mice in several ways, including high doses of cyclophosphamide and transfer of diabetic spleen cells to sublethally irradiated recipients. It has previously been established that transferred diabetes can be prevented by treatment with a nondepleting CD4 monoclonal antibody; however, we report herein that cyclophosphamide-induced diabetes also can be prevented using this antibody. The protection induced by CD4 monoclonal antibody to transferred diabetes is maintained for a long period after cessation of antibody treatment. However, cyclophosphamide can abrogate this induced tolerance and we report that this abrogation does not require new T cells. During the course of the experimental work described, we observed that the thymus had a suppressive effect on the expression of transferred disease. Mice that were depleted of their peripheral T-cells showed a doubling of the time for disease expression if they were euthymic, compared with thymectomized mice. PMID- 8405702 TI - Functional activity of plasma fibronectin in patients with diabetes mellitus. AB - Decreased wound healing and increased infection are major problems in patients with diabetes mellitus. Fibronectin plays a fundamental role in wound healing and acts as an opsonin for the phagocytosis of foreign antigens. The aim of this study was to ascertain the functional activity of plasma fibronectin from patients with diabetes mellitus. Initially, a modified Boyden chamber technique was used to measure cell migration on fibronectin purified from patient's plasma and an enzyme-linked immunosorbent assay was used to measure the binding of gelatin. A sandwich assay was developed that enabled the capture of fibronectin directly from patient's plasma without prior purification. With the use of a 96 well format, the binding of two different monoclonal antibodies could be compared simultaneously with the binding of gelatin and cell adhesion. In this way, differences in the function of particular domains of fibronectin from diabetic patients and control subjects could be measured. Results showed no difference between fibronectin from diabetic patients and control subjects with respect to the monoclonal antibodies binding in 1) the cell adhesion domain and 2) the heparin-binding domain. Furthermore, no detectable differences were noted with respect to cell adhesion, cell migration, or gelatin binding. These results suggest that diabetic patients receiving insulin treatment show no modulation of plasma fibronectin function, despite raised levels of circulating glucose. PMID- 8405703 TI - The role of glucose-6-phosphatase in the action of insulin on hepatic glucose production in the rat. AB - It has been suggested that regulation of glucose-6-phosphatase by insulin plays a role in the suppression of hepatic glucose production during feeding. We used hepatic glucose production (measured with the D-[3-3H]glucose infusion method) as an indicator of substrate flux through glucose-6-phosphatase in vivo. Compared with saline controls, insulin (7 mU.min-1 x kg-1, euglycemic clamp) suppressed hepatic glucose production virtually completely in both fasted (32.4 +/- 2.4 vs. 6.1 +/- 14 mumol.min-1 x kg-1) and fed (64.6 +/- 6.4 vs. 5.5 +/- 5.2 mumol.min-1 x kg-1) rats. Whereas hepatic glucose production was totally suppressed, [glucose 6-phosphate] in liver cytosol declined by only 27 and 35% in fasted and fed rats, respectively. Addition of hyperglycemia (10 mM) to the insulin infusion likewise fully suppressed hepatic glucose production (26.9 +/- 1.4 vs. -9 +/- 10 mumol.min 1 x kg-1 and 80.8 +/- 10.1 vs. -3.6 +/- 12.6 mumol.min-1 x kg-1 in fasted and fed rats, respectively), but [glucose-6-phosphate] again declined only modestly (21 and 27% in fasted and fed rats, respectively). This disproportionate suppression of hepatic glucose production could not be explained by cooperative substrate effects inasmuch as microsomal glucose-6-phosphatase isolated from saline- and insulin-treated rats followed Michaelis-Menten kinetics (Hill coefficient approximated 1). Acute insulin treatment of fasted rats in vivo did not reproducibly inhibit glucose-6-phosphatase activity assayed subsequently in isolated microsomes incubated in the absence of insulin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405704 TI - Hypoxia-induced sympathetic inhibition of the fetal plasma insulin response to hyperglycemia. AB - Large-for-delivery date babies, considered characteristic of diabetic pregnancy, are believed to result from fetal hyperinsulinemia. Paradoxically, infant birth weights tend to be low-for-delivery date in mothers with more severe diabetes. We tested the hypothesis that hypoxemia in such fetuses leads to sympathoadrenal stimulation and inhibition of insulin secretion; and, thus, produces a net reduction in the growth-promoting effects. Fetal sheep were prepared with chronic peripheral and adrenal cannulas. Fetal blood gases, lactate, norepinephrine, and epinephrine secretion rates; and plasma norepinephrine, glucose, and immunoreactive insulin concentrations were determined at 30-min intervals during a 2-h baseline period and a 4-h period of hyperglycemia divided into 2-h segments of hypoxemia (with and without alpha-blockade) and hyperoxia. Hypoxemia-hyperoxia sequences were varied randomly. Well-oxygenated fetuses responded to a threefold increase in glucose with a sixfold increase in plasma immunoreactive insulin. With hypoxemia, norepinephrine and epinephrine secretion were elevated and the insulin response was blocked. With hypoxemia and phentolamine blockade, the insulin response was enhanced with a 10-fold increase above baseline. In severe maternal diabetes with vascular disease or with poor control and very high glucose levels, the fetus is likely to be relatively hypoxemic. Our experiments suggest that in this situation, the fetal insulin response to hyperglycemia will be attenuated; this effect is mediated, at least partly, through sympathoadrenal stimulation. PMID- 8405705 TI - Alcohol intake impairs glucose counterregulation during acute insulin-induced hypoglycemia in IDDM patients. Evidence for a critical role of free fatty acids. AB - In this study, we assessed the effects of alcohol intake on glucose counterregulation in response to acute insulin-induced hypoglycemia in IDDM patients and in normal control subjects. Nine euglycemic IDDM patients and 9 normal control subjects were studied. After a baseline period, insulin (0.15 U/kg) was administered subcutaneously to induce hypoglycemia. Each IDDM patient was studied 3 times. In the first study, alcohol was orally administered as wine. In the second (control) study, water was administered instead of wine. In the third study, wine was given; however, a continuous infusion of heparin plus intralipid was administered to prevent the fall in plasma free fatty acid. Normal control subjects underwent only the alcohol and the control studies. In IDDM patients alcohol intake impairs, whereas in normal subjects it supports glucose counterregulation. Alcohol intake is associated with normal catecholamine responses in both IDDM diabetic patients and normal subjects. In both IDDM patients and normal subjects, hepatic glucose production in the recovery phase of the alcohol study was normal. Plasma glucose rate of disappearance was significantly increased by alcohol intake in IDDM (13.72 +/- 0.82 vs. 11.84 +/- 0.53 mumol.kg-1 x min-1; P < 0.05). Alcohol intake in both normal subjects and IDDM patients decreased plasma free fatty acid (267 +/- 22 vs. 156 +/- 20 microM; P < 0.01 and 356 +/- 29 vs. 96 +/- 12 microM; P < 0.01). We hypothesized that in IDDM patients, deficient glucose recovery during alcohol intake is the result of the ability of alcohol to depress lipolysis. PMID- 8405706 TI - Reduced sampling protocols in estimation of insulin sensitivity and glucose effectiveness using the minimal model in NIDDM. AB - Recent work in healthy subjects, the aged, and subjects with gestational diabetes or drug-induced insulin resistance using minimal model analysis of the tolbutamide-modified frequently sampled intravenous glucose tolerance test suggested that a reduced sampling regimen of 12 time points produced unbiased and generally acceptable estimates of insulin sensitivity (SI) and glucose effectiveness (SG) compared with a full sampling schedule of 30 time points. We have used data from 26 insulin-modified frequently sampled intravenous glucose tolerance tests in 21 subjects with NIDDM to derive and compare estimates of SI and SG from the full sampling schedule (SI(30), SG(30)) with those estimated from the suggested 12 time points (SI(12), SG(12)) and those estimated with the addition of a 25-min time point (SI(13), SG(13)). Percentage relative errors were calculated relative to the corresponding 30 time-point values. A statistically significant bias of 15% (97% confidence interval from 7.4 to 25.6%, interquartile range 25%) was introduced by the estimation of SI(12) but not SI(13) (1%, 97% confidence interval from -9.4 to 9.3%, interquartile range 21%). Results for SG(12) (-12%, 97% confidence interval from -46.7 to 1.2%, interquartile range 49%) and SG(13) (-5%, 97% confidence interval from -27.8 to 6.8%, interquartile range 37%) were statistically equivocal. The precision of estimation of SI(12), SG(12), and SG(13) measured by the interquartile range of the percentage relative errors was poor. The precision of determination measured by the median minimal model coefficient of variation was 18, 29, and 27% for SI(30), SI(12), and SI(13) and 9, 11, and 11% for SG(30), SG(12), and SG(13), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405707 TI - Cytoplasmic islet cell antibodies recognize distinct islet antigens in IDDM but not in stiff man syndrome. AB - Cytoplasmic islet cell antibodies are well-established predictive markers of IDDM. Although target molecules of ICA have been suggested to be gangliosides, human monoclonal ICA of the immunoglobulin G class (MICA 1-6) produced from a patient with newly diagnosed IDDM recognized glutamate decarboxylase as a target antigen. Here we analyzed the possible heterogeneity of target antigens of ICA by subtracting the GAD-specific ICA staining from total ICA staining of sera. This was achieved 1) by preabsorption of ICA+ sera with recombinant GAD65 and/or GAD67 expressed in a baculovirus system and 2) by ICA analysis of sera on mouse pancreas, as GAD antibodies do not stain mouse islets in the immunofluorescence test. We show that 24 of 25 sera from newly diagnosed patients with IDDM recognize islet antigens besides GAD. In contrast, GAD was the only islet antigen recognized by ICA from 7 sera from patients with stiff man syndrome. Two of these sera, however, recognized antigens besides GAD in Purkinje cells. In patients with IDDM, non-GAD ICA were diverse. One group, found in 64% of the sera, stained human and mouse islets, whereas the other group of non-GAD ICA was human specific. Therefore, mouse islets distinguish two groups of non-GAD ICA and lack additional target epitopes of ICA besides GAD. Longitudinal analysis of 6 sera from nondiabetic ICA+ individuals revealed that mouse-reactive ICA may appear closer to clinical onset of IDDM in some individuals. Mouse-reactive ICAs, however, remained absent in 36% of the patients at diagnosis of IDDM. PMID- 8405708 TI - The pharmacodynamics and activity of intranasally administered insulin in healthy male volunteers. AB - The objective of the study was to assess the bioavailability and absorption dynamics of intranasal insulin (with di-decanoyl-alpha-phosphatidylcholine, DDPC, as absorption enhancer) in two potencies (U200 and U500). Toward this aim, the euglycemic clamp technique combined with somatostatin (100 micrograms/h) was used. Insulin was administered to 12 healthy males: 5 IU intravenously (20-min infusion); 10 IU subcutaneously; 50 IU (U200) and 50 IU, 100 IU, and 150 IU (U500) intranasally. Peak insulin levels (mean +/- SD) were reached at 17.9 +/- 2.6, 77.9 +/- 38.3, 23.3 +/- 5.4, 25.4 +/- 8.4, 26.2 +/- 8.3, and 27.5 +/- 5.8 min, respectively. For the 50 IU dose, peak glucose requirements during the clamp and time to peak were not significantly different for U200 and U500: 548.8 +/- 279.5 vs. 452.4 +/- 232.9 mg/min and 41.3 +/- 16.2 vs. 51.5 +/- 29.9 min, respectively. Compared with intravenous insulin, the bioavailability calculated from the total area under the insulin curve was 13.2% (95% confidence interval 7.9, 21.9) and 8.8% (95% confidence interval 5.6, 13.8), and compared with subcutaneous insulin, the bioavailability was 14.8% (95% confidence interval 8.7, 25.2) and 9.9% (95% confidence interval 6.4, 15.4) for the U200 and U500 preparations, respectively. An apparent nonlinear dose-dependent relation was found for the U500 potency. The within-subject variability of the areas under the curves of plasma insulin after the administration of 100 IU was 43.6% (range 20.7 85.7). In conclusion, this nasal insulin preparation has promising absorption and action profiles in both potencies, which makes it suitable for further exploration of clinical applications. PMID- 8405709 TI - Endothelial cell-binding properties of lymphocytes infiltrated into human diabetic pancreas. Implications for pathogenesis of IDDM. AB - In IDDM, mononuclear cells accumulate in the islets of Langerhans and destroy insulin-producing beta-cells. To study the mechanisms that control extravasation of circulating mononuclear cells into the pancreas, we examined the phenotype of vascular endothelium of the pancreas, propagated a T-cell line from pancreatic islets at the onset of the disease and compared endothelial binding of this cell line in vitro to vascular endothelium in different body regions. The adhesion molecules expressed on the resulting T-cell line and the functional binding capacity of these cells to the endothelium of the normal and diabetic pancreas, mucosa-associated lymphatic tissues, and regional and peripheral lymph nodes were studied. We present evidence of pancreatic endothelial activation in diabetes, leading to endothelial morphology typical for HEVs and accompanying local increase in extravasation of mononuclear cells into the pancreas. Endothelial cell binding experiments with the T-cell line showed strong adherence of the cells to the endothelium of diabetic pancreas and mucosal lymphoid tissue. The cell line was uniformly CD4-positive, TCR V beta 5.1-positive, LFA-1-positive (CD 11a/CD18), VLA-4 alpha-positive (CD 49d), and CD 44-positive but negative for L selectin (peripheral lymph node homing receptor). The pancreatic or control cell lines showed no binding to vessels of normal pancreas, and the binding of the pancreatic cell line to the endothelium of peripheral lymph node was weak. Our results suggest that lymphocyte-endothelial cell interactions are important for the accumulation of inflammatory mononuclear cells into the pancreas and imply that lymphocytes derived from the mucosal lymphoid tissue may be involved in the pathogenesis of IDDM. PMID- 8405710 TI - Quantification of the relationship between insulin sensitivity and beta-cell function in human subjects. Evidence for a hyperbolic function. AB - To determine the relationship between insulin sensitivity and beta-cell function, we quantified the insulin sensitivity index using the minimal model in 93 relatively young, apparently healthy human subjects of varying degrees of obesity (55 male, 38 female; 18-44 yr of age; body mass index 19.5-52.2 kg/m2) and with fasting glucose levels < 6.4 mM. SI was compared with measures of body adiposity and beta-cell function. Although lean individuals showed a wide range of SI, body mass index and SI were related in a curvilinear manner (P < 0.0001) so that on average, an increase in body mass index was associated generally with a lower value for SI. The relationship between the SI and the beta-cell measures was more clearly curvilinear and reciprocal for fasting insulin (P < 0.0001), first-phase insulin response (AIRglucose; P < 0.0001), glucose potentiation slope (n = 56; P < 0.005), and beta-cell secretory capacity (AIRmax; n = 43; P < 0.0001). The curvilinear relationship between SI and the beta-cell measures could not be distinguished from a hyperbola, i.e., SI x beta-cell function = constant. This hyperbolic relationship described the data significantly better than a linear function (P < 0.05). The nature of this relationship is consistent with a regulated feedback loop control system such that for any difference in SI, a proportionate reciprocal difference occurs in insulin levels and responses in subjects with similar carbohydrate tolerance. We conclude that in human subjects with normal glucose tolerance and varying degrees of obesity, beta-cell function varies quantitatively with differences in insulin sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405711 TI - Transforming growth factor-beta 1 enhances glomerular collagen synthesis in diabetic rats. AB - In vitro glomerular collagen synthesis and its response to various concentrations of transforming growth factor-beta 1 were studied in normal and diabetic rats. TGF-beta 1 increased collagen synthesis in normal glomeruli in a dose-dependent manner up to 5 ng/ml. Concentrations > 5 ng/ml showed a gradual decline in collagen synthesis. Basal collagen synthesis was increased in diabetic glomeruli, but addition of TGF-beta 1 had no effect. Antibody to TGF-beta 1 prevented this increase in collagen synthesis. Both circulating TGF-beta 1 concentration and its glomerular expression of mRNA were higher in diabetic than in normal rats. Although addition of TGF-beta 1 did not increase synthesis in vitro, the absence of an effect is consistent with downregulation of its receptors attributable to the high circulating levels. This study clearly indicates a regulatory role of TGF-beta 1 in renal glomerular collagen synthesis in the normal rat, and suggests a possible causal role for enhanced collagen synthesis in the diabetic rat. PMID- 8405712 TI - Cloning and functional expression of the human islet GLP-1 receptor. Demonstration that exendin-4 is an agonist and exendin-(9-39) an antagonist of the receptor. AB - A complementary DNA for a glucagon-like peptide-1 receptor was isolated from a human pancreatic islet cDNA library. The isolated clone encoded a protein with 90% identity to the rat receptor. In stably transfected fibroblasts, the receptor bound [125I]GLP-1 with high affinity (Kd = 0.5 nM) and was coupled to adenylate cyclase as detected by a GLP-1-dependent increase in cAMP production (EC50 = 93 pM). Two peptides from the venom of the lizard Heloderma suspectum, exendin-4 and exendin-(9-39), displayed similar ligand binding affinities to the human GLP-1 receptor. Whereas exendin-4 acted as an agonist of the receptor, inducing cAMP formation, exendin-(9-39) was an antagonist of the receptor, inhibiting GLP-1 induced cAMP production. Because GLP-1 has been proposed as a potential agent for treatment of NIDDM, our present data will contribute to the characterization of the receptor binding site and the development of new agonists of this receptor. PMID- 8405713 TI - Meticulous prevention of hypoglycemia normalizes the glycemic thresholds and magnitude of most of neuroendocrine responses to, symptoms of, and cognitive function during hypoglycemia in intensively treated patients with short-term IDDM. AB - To test the hypothesis that hypoglycemia unawareness is largely secondary to recurrent therapeutic hypoglycemia in IDDM, we assessed neuroendocrine and symptom responses and cognitive function in 8 patients with short-term IDDM (7 yr) and hypoglycemia unawareness. Patients were assessed during a stepped hypoglycemic clamp, before and after 2 wk and 3 mo of meticulous prevention of hypoglycemia, which resulted in a decreased frequency of hypoglycemia (0.49 +/- 0.05 to 0.045 +/- 0.03 episodes/patient-day) and an increase in HbA1c (5.8 +/- 0.3 to 6.9 +/- 0.2%) (P < 0.05). We also studied 12 nondiabetic volunteer subjects. At baseline, lower than normal symptom and neuroendocrine responses occurred at lower than normal plasma glucose, and cognitive function deteriorated only marginally during hypoglycemia. After 2 wk of hypoglycemia prevention, the magnitude of symptom and neuroendocrine responses (with the exception of glucagon and norepinephrine) nearly normalized, and cognitive function deteriorated at the same glycemic threshold and to the same extent as in nondiabetic volunteer subjects. At 3 mo, the glycemic thresholds of symptom and neuroendocrine responses normalized, and surprisingly, some of the responses of glucagon recovered. We concluded that hypoglycemia unawareness in IDDM is largely reversible and that intensive insulin therapy and a program of intensive education may substantially prevent hypoglycemia and at the same time maintain the glycemic targets of intensive insulin therapy, at least in patients with IDDM of short duration. PMID- 8405714 TI - Sombre reading. PMID- 8405715 TI - Non-accidental head injury, with particular reference to whiplash shaking injury and medico-legal aspects. PMID- 8405716 TI - Influence of different sitting positions and abduction orthoses on leg muscle activity in children with cerebral palsy. AB - A surface electromyogram (EMG) was recorded from four leg muscles to measure the effects of various seat inclinations and an abduction orthosis on eight children with cerebral palsy and controls during performance of an upper-extremity task and while listening to a story. EMG responses were lowest in the forward-leaning and horizontal positions with the abduction orthosis, and highest in reclined and horizontal positions without the orthosis for both groups of children during the performance of an upper-extremity task. While listening to a story, there was no median EMG activity in any muscles of the controls, but there were wide variations in those of the children with cerebral palsy. The results indicate that the use of an abduction orthosis and horizontal and forward-leaning seats decrease lower-extremity muscle activity, and so it is possible that it might also improve upper-extremity function. PMID- 8405717 TI - Children with deficits in attention, motor control and perception (DAMP) almost grown up: general health at 16 years. AB - One hundred and one children (56 with and 45 without deficits in attention, motor control and perception (DAMP)), originally diagnosed at six or seven years of age and belonging to a representative cohort of children from the general population were followed up at 16 and 17 years of age. There was a significant excess of substance abuse, fractures and other accidents in the DAMP group than among controls, in addition to more motor co-ordination problems, clumsiness, and height and weight problems. Mean complex visual reaction time was significantly longer in the DAMP group, accounted for by the number of boys in the group. It appears that 10-year outcome for children who had attention problems and clumsiness during the preschool period is considerably poorer than for those who did not have such problems. PMID- 8405718 TI - Anorectal function of children with neurological problems. I: Spina bifida. AB - Anorectal function was assessed with anorectal manometry in 45 children with spina bifida (21 girls and 24 boys, mean age 11 1/2 years). 24 enuretic children served as controls. The pressure in the first and second centimeters of the anal canal was lower among index children than controls and also lower among those with high spinal lesions compared with those with low lesions. Rectal activity (rectal sensation and trace appearance) during rectal distension appeared to be reduced among index children; sensation was particularly poor among those with high spinal lesions. Manometry may be useful in children with spina bifida as it provides a clearer understanding of sphincter function and leads to a more rational approach to the management of bowel problems. PMID- 8405719 TI - Anorectal function of children with neurological problems. II: cerebral palsy. AB - In response to the frequent complaint of difficulties with defecation experienced by children with cerebral palsy, 34 children (13 girls and 21 boys, mean age 10 years) with cerebral palsy were investigated by questionnaire and anorectal manometry. 24 enuretic children served as controls for the anorectal manometry. Constipation affected 26 of 29, defecation distress eight and faecal incontinence 16 of the index children, but incontinence was mild in most cases. Index children had a low resting pressure in the first centimetre of the anal canal, slow anal rhythmical activity and a pressure increase in the first centimetre during maximum rectal distension. These findings suggest anal sphincter and/or pelvic floor muscle incoordination, but no evidence of abnormal rectal function. The authors conclude that surgical intervention was not indicated for the index children, but that medical treatment could be improved. PMID- 8405720 TI - Motor function of infants with athetoid cerebral palsy. AB - The motor function of 35 children with athetoid cerebral palsy was examined retrospectively using videotape recordings made at five to eight months of age. Many infants showed asymmetric tonic neck, Moro and Galant reflexes. Movements shown to be difficult included: keeping a symmetric supine posture, isolated movements of the hips and knees, forward extension of the upper extremity, extension of neck and trunk in the prone position and in ventral suspension, flexion of the neck in the traction response, and weight support by the upper extremities. Asymmetric or excessive opening of the mouth was present in all infants. The grade of difficulty for each posture and movement might reflect subsequent motor disability at three years of age. PMID- 8405721 TI - Congenital livedo reticularis and recurrent stroke-like episodes. AB - Three children with pronounced livedo reticularis present since birth (cutis marmorata-telangiectasia congenita) have been followed to the ages of eight, 17 and 21 years. During childhood they developed frequent recurrent transient stroke like hemipareses, affecting either side of the body, associated with ipsilateral pain, headache, visual symptoms, dysphasia, fits and confusion. Intellectual failure and, in one, progressive spasticity have followed. Attacks were more frequent in winter. Other problems have included abnormal peripheral vascular responses to temperature change, gastro-intestinal bleeding, glaucoma, local tissue hypertrophy and, in the two older patients, renal involvement with hypertension. Their condition represents a form of congenital vasculopathy. Anticonvulsants, anti-migraine agents, anti-platelet drugs and flunarizine have been ineffective. Nifedipine prevented further attacks in one patient and reduced attacks in another, but has not helped the third child. Adequate clothing and warmth may also be important. PMID- 8405722 TI - Ictal cortical blindness: a case report and review of the literature. AB - Acute cortical blindness as an ictal phenomenon has seldom been reported in the literature. The authors describe a seven-year-old boy who experienced several episodes of complete visual loss, accompanied by gastro-intestinal symptoms and a sensation of fright, but with preservation of consciousness. These episodes ended abruptly with visual recovery and no postictal phenomena. CT brain scan was normal and interictal EEG showed bi-occipital epileptiform activity. This case fits the definition of true ictal blindness. A literature review of seizure related blindness is presented. PMID- 8405723 TI - Club foot. PMID- 8405724 TI - Videofluoroscopy in the assessment of feeding disorders. PMID- 8405725 TI - Postictal plasma ACTH in fifth-day fits of the newborn infant. PMID- 8405726 TI - Overlooked? PMID- 8405727 TI - Mismatch negativity in the neurophysiologic/behavioral evaluation of auditory processing deficits: a case study. AB - The subject of this case report is an 18-year-old woman with grossly abnormal auditory brain stem response (ABR), normal peripheral hearing, and specific behavioral auditory processing deficits. Auditory middle latency responses (MLRs) and cortical potentials N1, P2, and P300 were intact. The mismatch negativity (MMN) was normal in response to certain synthesized speech stimuli and impaired to others--consistent with her behavioral discrimination of these stimuli. Behavioral tests of auditory processing were consistent with auditory brain stem dysfunction. A neuropsychological evaluation revealed normal intellectual and academic performance. The subject was in her first year of college at the time of the evaluation. This case study is important because: (1) Although there have been several reports of absent/abnormal ABR with preserved peripheral hearing and deficits in auditory processing, little is known about the specific nature of the auditory deficits experienced by these individuals. Such information may be valuable to the clinical management of patients with this constellation of findings. (2) Of interest is the information that the mismatch negativity (MMN) cortical event-related potential can bring to the evaluation of patients with auditory processing deficits. The MMN reflects central auditory processing of small acoustic differences and may provide an objective measure of auditory discrimination. (3) From a theorectical standpoint, a patient with neural deficits affecting specific components of the auditory pathway provides insight into the relationship between evoked potentials and physiological mechanisms of auditory processing. How do various components of the auditory pathway contribute to speech discrimination? How might evoked potentials reflect the processes underlying the neural coding of specific features of speech stimuli such as timing and spectral cues? PMID- 8405728 TI - The influence of personality-related factors upon consultation for two different "marginal" organic pathologies with and without reports of auditory symptomatology. AB - Obscure Auditory Dysfunction (OAD) is explained by a combination of hearing related deficits and personality factors Saunders & Haggard (1992). In this study, we determine which factors are associated specifically with OAD and which are associated with the seeking of medical attention in general. We achieved this by obtaining a second patient group with a parallel syndrome to OAD, called "chronic pelvic pain without obvious organic pathology" (CPPWOOP). CPPWOOP patients complain of lower abdominal pain that is not explainable by conventional medical tests. Fifteen CPPWOOPs underwent the OAD test battery. For the analyses they were retrospectively matched to 15 of the original OADs and their matched controls. The three groups were compared by analysis of variance and Kruskall Wallis analyses. The CPPWOOPs and controls performed significantly better than OADs on hearing-related variables, but did not differ from each other, whereas the OADs and CPPWOOPs were significantly more anxious than the controls, but did not differ from each other. We conclude that anxiety-related traits are associated with the seeking of medical attention in general, whereas the hearing related deficits we measured are associated specifically with OAD. Anxiety related traits should, therefore, be considered when dealing with marginal pathologies, but in depth investigation may also reveal an organic basis; therefore, patients should not be dismissed as simply neurotic. PMID- 8405729 TI - The hamster's auditory brain stem response as a function of stimulus intensity, tone burst frequency, and hearing loss. AB - Young adult hamsters were evaluated for normal variations in the auditory brain stem response (ABR) and for the effects of hearing loss. Normal variations in the ABR's latencies and amplitudes were defined for both click and tone burst stimuli over a broad range of intensities. As stimulus intensity decreased from 100 to 15 dB pe SPL, the ABR latencies were prolonged, the P4-P1 interpeak latencies varied in a complex manner, and amplitudes decreased. As tone burst frequency decreased from 8000 to 4000 and then to 2000 Hz, ABR latencies were prolonged, amplitudes decreased, and thresholds increased. The P4-P1 interpeak latencies were also influenced by tone burst frequency, with the longest interpeak latencies occurring in response to the 2000 Hz tone bursts. Data from two hamsters with hearing loss were compared to the normal data to illustrate how the ABR can be used to describe experimentally induced hearing impairment. Hearing impairment was induced by rearing animals in ambient noise. These animals showed elevated ABR thresholds at all three frequencies and upward displacements in the ABR's latency-intensity profiles. PMID- 8405730 TI - Distortion-product and click-evoked otoacoustic emissions of preterm and full term infants. AB - Full-term and preterm infants were evaluated with click-evoked and distortion product otoacoustic emissions (CEOEs and DPOEs). The CEOEs and DPOEs recorded from each individual ear were analyzed by calculating the root-mean-square levels within half-octave bands. The fail criterion of the OE tests was that the half octave RMS DPOE or CEOE levels of an ear under test were below the 10th percentile of full-term newborns in two or more bands. The DPOE data were collected from 118 ears of 61 premature babies; 80 (68%) ears passed the DPOE test, 30 (25%) ears without middle ear effusions failed the test, and 8 (7%) ears with effusions also failed. The CEOE data were collected from 128 ears of 65 premature babies; 102 (80%) ears passed the CEOE test, 18 (14%) ears without middle ear effusions failed the test, and 8 (6%) ears with effusions also failed. In 23 of 80 ears (29%) that passed the DPOE test and in 23 of 102 ears (23%) that passed the CEOE test, RMS OE levels of preterm infants were above the 90th percentile of full-term newborns. The analyses of the combined DPOE and CEOE data obtained from a group of 25 ears of full-term newborns and from a group of 72 ears of preterm babies showed statistically significant correlations between the DPOE and CEOE root-mean-square levels in each of the half-octave bands in the 1.4 to 4 kHz region. For 42 preterm infants tested with auditory brain stem response (ABR), specificity was 86% for CEOE and 74% for DPOE. All infants who failed the ABR also failed OE tests. To the best of our knowledge, this study is the first using combined DPOEs, CEOEs, and ABRs for preterm babies. It showed the feasibility of DPOEs and CEOEs for this population. PMID- 8405731 TI - Prediction of benefit from linear hearing aids in nonreverberant listening environments. AB - Hearing aid benefit, defined in terms of improved speech intelligibility, was measured for 16 elderly hearing-impaired subjects. Twelve conditions were tested, simulating a range of daily situations from typical home environments to moderate sized social gatherings, and assuming a small talker-listener distance (thus maintaining essentially nonreverberant listening conditions). Each subject was fitted with the same type of programmable hearing aid. The goals were to develop a model for the prediction of benefit based on hearing loss, listening environment, and amplification variables, and to assess the potential accuracy of the model. Two models were developed using multiple linear regression analyses. The prefitting model used data that would be available before a hearing aid fitting, that is, audiogram and listening environment data. This model, although potentially useful as a counseling tool, was relatively inaccurate. Six of the 16 subjects yielded benefit data that were consistently different from the model's predictions. The postfitting model used information that could be obtained during a hearing aid fitting about audibility changes resulting from amplification. This model produced more accurate, but still imperfect predictions of benefit. Benefit obtained by three subjects deviated substantially from the predictions of the postfitting model. It was concluded that a model producing fairly accurate benefit predictions must encompass additional variables beyond those considered here. Nevertheless, these models may be useful for prediction of typical benefit for potential hearing aid wearers. PMID- 8405732 TI - The influence of hearing aid cost on perceived benefit in older adults. AB - Self-report hearing handicap scales are gaining widespread acceptance among clinicians as a technique for quantifying hearing aid benefit. Both auditory and nonauditory (e.g., personality, health, motivation) factors contribute to an individual's response to hearing loss and ultimately to perceived hearing aid benefit. Among the recognized extra-audiological variables, financial concerns may influence a hearing aid user's expectations regarding hearing aid benefit. The present study compared self-perceived benefit in 26 insured and 26 uninsured new hearing aid wearers using the Hearing Handicap Inventory for the Elderly (HHIE). HHIE scores at 3 weeks and 6 mo postfitting were equivalent between groups. The findings suggest that the HHIE may be used as an index of hearing aid benefit without concern that the source of funds required to purchase amplification may act as an uncontrolled variable. PMID- 8405733 TI - Loudness balance between acoustic and electric stimulation by a patient with a multichannel cochlear implant. AB - Estimates of loudness balance were obtained for acoustically and electrically presented 250 Hz sine signals from a patient who uses the Ineraid multichannel cochlear implant. Acoustic and electric loudness matching was possible because the patient evidenced a 25 dB HL threshold at 250 Hz in his nonimplanted ear. The level of the electrical stimulus in microamperes required for a balance of loudness grew linearly with equal increments in decibels for the acoustic stimulus. These data, in concert with the very limited data from previous studies, provide a rationale for using a logarithmic transformation of acoustic to electric intensity in signal processors for cochlear implants. PMID- 8405734 TI - Within-subject versus between-subject variability comparison. PMID- 8405735 TI - Diabetes mellitus due to viruses--some recent developments. AB - Many different viruses belonging to several genera have the potential to damage beta cells. The mechanisms they employ are varied, and infection may result in either a direct destruction of islets and rapid insulin deficiency, or in a more gradual loss of functioning islets with the onset of diabetes many years later. Several case histories involving extensive cytolysis of beta cells can be directly linked to viral infection, whilst an example of diabetes occurring many years after viral infection is found in individuals who had a congenital infection with rubella virus. Here, the virus induces an autoimmune reaction against beta cells. Autoimmune phenomena have also been observed in islets following infections with viruses other than rubella, and thus activation of autoimmune mechanisms leading to beta-cell destruction may be a relatively frequent occurrence. Recent evidence shows that picornaviruses are not exclusively lytic, and can induce more subtle, long-term changes in beta cells, which may be important in the aetiology of diabetes. The exact mechanisms involved are not known, but it is clear that several viruses can directly inhibit insulin synthesis and induce the expression of other proteins such as interferons, and the HLA antigens. Strain differences in viruses are important since not all variants are tropic for the beta cells. Several laboratories are in the process of identifying the genetic determinants of tropism and diabetogenicity, especially amongst the Coxsackie B (CB) virus group. The sequence of one such diabetogenic CB4 strain virus has been determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405736 TI - Glucose transporter gene expression in rat conceptus during high glucose culture. AB - We investigated the expression of glucose transporter genes and protein in embryo and yolk sac during organogenesis and the regulation of glucose transporters during culture in hyperglycaemic media. Erythrocyte-type glucose transporter (GLUT 1) and brain-type glucose transporter (GLUT 3) mRNA were expressed in embryo and yolk sac. The expression of GLUT-1 and GLUT-3 mRNA was abundant on day 9-11 and day 9-10 in the embryo, respectively, and day 9-14 and day 10-11 in the yolk sac, respectively. The levels of GLUT-1 protein in the embryo increased in parallel with the expression of GLUT-1 mRNA during the corresponding period. Immunohistochemical staining of GLUT-1 protein was found principally in the neuroepithelial cells surrounding the neural tube in the embryo on day 10 and appeared in the microvessels surrounding the neural tube after day 12. To test whether the expression of glucose transporter genes and protein was suppressed during hyperglycaemia, conceptuses were cultured in high glucose medium. The abundant expression of GLUT-1 protein was not decreased during culture in high glucose media for 24 h (day 9-10) and was only down-regulated by prolonged exposure to this media for 48 h (day 9-11). We have demonstrated the predominant expression of the high affinity glucose transporter (GLUT 1 and GLUT 3) genes and (GLUT 1) protein in embryo during the early period of organogenesis. The persistently abundant expression of glucose transporter during the critical period of neural tube formation (day 9-10) even in the presence of hyperglycaemia may explain one of the mechanism of increased glucose flux into the neuroepithelium, which may lead to neural tube defects. PMID- 8405737 TI - Alloxan cytotoxicity is highly potentiated by plasma membrane- and lysosomal associated iron--a study on a model system of cultured J-774 cells. AB - Pancreatic islet beta cells, and some other cell types, are sensitive to the damaging effects of alloxan. The mechanisms behind the cytotoxicity have not been fully elucidated, although they are considered to be mediated by the formation and effects of reactive oxygen metabolites. In the present study, the cytotoxic effects of alloxan/cysteine at high and low concentrations were investigated on a model system of cultured J-774 cells. Viability was estimated by the trypan blue dye exclusion test, plasma membrane permeability by a modified microfluorometric fluorescein diacetate technique and lysosomal membrane stability by a microfluorometric acridine orange method. The results showed: (a) hydrogen peroxide, readily diffusing through cellular membranes and produced extracellularly in large amounts by alloxan/cysteine at high concentrations, enters the secondary lysosomes if not previously degraded by cellular anti oxidant systems. Intralysosomal Fenton reactions, with the formation of hydroxyl radicals, may be induced provided catalytically active lysosomal iron is present. This would result in lysosomal membrane damage followed by leakage of lysosomal contents to the cell sap and cell degeneration. (b) Alloxan/cysteine at low concentrations induced production of superoxide and hydrogen peroxide in low amounts which caused almost no lysosomal damage and appeared to be non-toxic unless there was some plasma membrane-associated iron. Consequently, cells initially allowed to endocytose iron during culture, or briefly exposed to iron just before exposure to alloxan and cysteine, showed greatly enhanced sensitivity. In this case iron, in combination with superoxide and hydrogen peroxide, is believed to give rise to plasma membrane-associated hydroxyl radical production (Fenton reaction) with resultant loss of membrane integrity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405738 TI - Suppression of diabetes mellitus in the non-obese diabetic (NOD) mouse by an autoreactive (anti-I-Ag7) islet-derived CD4+ T-cell line. AB - The non-obese diabetic (NOD) mouse is a spontaneous model of human insulin dependent diabetes mellitus. Both CD4+ and CD8+ T cells infiltrate the pancreatic islets of NOD mice prior to beta-cell destruction. T-cell lines isolated from the islets of NOD mice are tools for studying the pathogenesis of insulin-dependent diabetes mellitus. During attempts to generate such lines we isolated an autoreactive CD4+ T-cell line, designated C2, from the 'insulitis' lesion of a 20 week-old female non-diabetic NOD/WEHI mouse. Islet T cells were propagated by the addition of interleukin-2 and reexposure every 2 weeks to whole NOD islets and irradiated NOD spleen cells as antigen presenting cells. C2 cells proliferated up to 100-fold upon exposure to NOD antigen presenting cells but did not respond to whole NOD islets or antigen presenting cells from allogeneic mouse strains. Proliferation of C2 cells to NOD antigen presenting cells was blocked by a monoclonal antibody against the unique class II MHC molecule of NOD, I-Ag7. In response to NOD antigen presenting cells, C2 cells secreted interferon-gamma, tumour necrosis factor-alpha and interleukin-6 but no detectable interleukin-2, interleukin-4 or interleukin-10, a pattern of cytokine secretion more characteristic of Th1 CD4 cells. C2 cells displayed significant cytotoxicity in a redirected lysis assay. To explore a possible role for autoreactive T cells in the pathogenesis of autoimmune diabetes, C2 cells were injected i.v. into female NOD mice that had received cyclophosphamide to accelerate development of diabetes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405739 TI - Deficient activity of FAD-linked glycerophosphate dehydrogenase in islets of GK rats. AB - In pancreatic islet extracts of rats with hereditary non-insulin-dependent diabetes mellitus (GK rats), the activity of the mitochondrial FAD-linked glycerophosphate dehydrogenase, as measured by either a radioisotopic or colorimetric procedure, only represented 30 to 40% of that found in control rats. This decrease in enzymic activity was not attributable to any sizeable change in either islet DNA content or the relative contribution of insulin-producing beta cells to total islet mass. It contrasted with a normal activity of other mitochondrial dehydrogenases and hexokinase isoenzymes. It coincided, however, with an increased activity of glutamate-pyruvate transaminase, as already observed in adult rats injected with streptozotocin during the neonatal period. The decreased activity of islet FAD-linked glycerophosphate dehydrogenase also contrasted with an increased activity of the same enzyme in the liver of GK, as compared to control rats. In the light of these findings and recent metabolic data collected in intact islets of GK rats, it is proposed that a deficiency of beta-cell FAD-linked glycerophosphate dehydrogenase, the key enzyme of the glycerol phosphate shuttle, may represent a cause of inherited non-insulin dependent diabetes. PMID- 8405740 TI - Impact of initial treatment on renal function in newly-diagnosed type 2 (non insulin-dependent) diabetes mellitus. AB - The impact of improved glycaemic control on renal function in newly-presenting Type 2 (non-insulin-dependent) diabetic patients has not been adequately researched. Consequently, glomerular filtration rate and effective renal plasma flow and urinary albumin excretion rates were determined in 76 subjects (age (mean (SD)): 54 (9.5) years; 50 male) of an original cohort of 110 newly presenting normotensive non-proteinuric Type 2 diabetic patients following 6 months treatment with diet alone (n = 42) or with oral hypoglycaemic agents (n = 34). Significant reductions were observed in (presentation vs 6 months): body mass index (p < 0.001); fasting plasma glucose (p < 0.001); glycated haemoglobin (HbA1) (p < 0.001); systolic blood pressure (p < 0.01); and diastolic blood pressure (p < 0.001). Glomerular filtration rate declined from 117 (22) to 112 (21) ml.min-1 (p < 0.01), with unchanged effective renal plasma flow (534 (123) vs 523 (113) ml.min-1) and filtration fraction (22.4 (3.0) vs 21.8 (3.4)%). Albumin excretion rate (median (range)) declined from 1.1 (0.1-34.7) to 0.5 (0.1 29.9) micrograms.min-1 (p < 0.01). Changes in glomerular filtration rate (delta values) were inversely correlated with presentation values (p < 0.001), and positive relationships were observed with delta effective renal plasma flow (p < 0.01), and delta glycated haemoglobin (p < 0.05). Type 2 diabetic patients with glomerular filtration rate values at presentation over 120 ml.min-1 demonstrated significant reduction in glomerular filtration rate (n = 31; p < 0.001), whilst those with original values less than 120 ml.min-1 remained unchanged (n = 45).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405741 TI - Normalization of fasting hyperglycaemia by exogenous glucagon-like peptide 1 (7 36 amide) in type 2 (non-insulin-dependent) diabetic patients. AB - Glucagon-like peptide 1 (GLP-1) (7-36 amide) is a physiological incretin hormone that is released after nutrient intake from the lower gut and stimulates insulin secretion at elevated plasma glucose concentrations. Previous work has shown that even in Type 2 (non-insulin-dependent) diabetic patients GLP-1 (7-36 amide) retains much of its insulinotropic action. However, it is not known whether the magnitude of this response is sufficient to normalize plasma glucose in Type 2 diabetic patients with poor metabolic control. Therefore, in 10 Type 2 diabetic patients with unsatisfactory metabolic control (HbA1c 11.6 +/- 1.7%) on diet and sulphonylurea therapy (in some patients supplemented by metformin or acarbose), 1.2 pmol x kg-1 x min-1 GLP-1 (7-36 amide) or placebo was infused intravenously in the fasting state (plasma glucose 13.1 +/- 0.6 mmol/l). In all patients, insulin (by 17.4 +/- 4.7 nmol x 1-1 x min; p = 0.0157) and C-peptide (by 228.0 +/ 39.1 nmol x 1-1 x min; p = 0.0019) increased significantly over basal levels, glucagon was reduced (by -1418 +/- 308 pmol x 1-1 x min) and plasma glucose reached normal fasting concentrations (4.9 +/- 0.3 mmol/l) within 4 h of GLP-1 (7 36 amide) administration, but not with placebo. When normal fasting plasma glucose concentrations were reached insulin returned towards basal levels and plasma glucose concentrations remained stable despite the ongoing infusion of GLP 1 (7-36 amide). Therefore, exogenous GLP-1 (7-36 amide) is an effective means of normalizing fasting plasma glucose concentrations in poorly-controlled Type 2 diabetic patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405742 TI - Diabetes mellitus and male sexual function: a controlled study. AB - There is an extensive clinical literature on the erectile disorders of diabetic men but a paucity of controlled studies that have taken into account the effects of age, concurrent illnesses and medication on sexual function. This investigation was carried out on 40 diabetic men free from other illness or drugs that could affect sexual capacity and 40 age-matched healthy control subjects. Each subject and his female partner underwent semistructured interviews and the men had comprehensive medical evaluations and polygraphic assessment of sleep and nocturnal penile tumescence in the sleep laboratory during three nights. In comparison to control subjects, diabetic patients reported significant decreases in sexual desire, subjective arousal, erectile capacity, coital frequency and sexual satisfaction. The diabetic group also had significant decrements in duration of rapid eye movement sleep and in frequency, duration and degree of nocturnal penile tumescent episodes. There were no differences between Type 1 (insulin-dependent) and Type 2 (non-insulin dependent) diabetic patients in prevalence of sexual problems or in nocturnal tumescent measures. Significant relations were observed between lack of metabolic control, diabetic complications and impaired nocturnal tumescence. Sexually non-dysfunctional diabetic men had significant nocturnal penile tumescence abnormalities. Diabetic men without coital failures may have a subclinical impairment in erectile function which, although of not significant magnitude to interfere with penetration, is reflected in nocturnal penile tumescent measures. This result raises a note of caution in the interpretation of the nocturnal penile tumescence test for the differential diagnosis of diabetic erectile impotence. PMID- 8405743 TI - Impact of injection sites for soluble insulin on glycaemic control in type 1 (insulin-dependent) diabetic patients treated with a multiple insulin injection regimen. AB - The absorption rate of rapid acting (soluble) insulin is slow from the subcutaneous tissue of the thigh compared to intramuscular injection into the thigh and s.c. injection into the abdominal wall. The aim of the study was to evaluate the impact of soluble insulin injected either intramuscularly into the thigh (IMT), s.c. into the abdominal wall (SCA) or s.c. into the thigh (SCT) on glycaemic control in Type 1 (insulin-dependent) diabetic outpatients treated with the basal bolus insulin delivery regimen. Fifty-five, C-peptide negative Type 1 diabetic outpatients were included in a randomised 3-month intervention study. The insulin doses were adjusted frequently by blinded observers based on the patients' self-monitored blood glucose values and reported hypoglycaemic episodes. The serum fructosamine value was within normal limits in three patients in the IMT group, in six patients in the SCA group and in none of the patients in the SCT group following the intervention period (p < 0.01). However, the difference in mean serum fructosamine values did not reach statistical significance (IMT: 1.24 mmol/l (95% confidence interval; 1.17 to 1.31), SCA: 1.25 mmol/l (1.18 to 1.32), SCT: 1.34 mmol/l (1.26 to 1.41), (p = 0.09). Blood glucose excursions were larger in the SCT group than in the SCA and IMT group from post lunch to pre-dinner measurements and from pre- to post-dinner measurements. A higher number of measured low nocturnal blood glucose values (less than 4 mmol/l) was observed in the SCT group (34 of 85) than in the IMT (14 of 64) and SCA (21 of 81) group (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405744 TI - Differential interleukin-1 receptor antagonism on pancreatic beta and alpha cells. Studies in rodent and human islets and in normal rats. AB - The monokines interleukin-1 alpha and -beta have been implicated as effector molecules in the immune-mediated pancreatic beta-cell destruction leading to insulin-dependent diabetes mellitus. Here we investigated the effects of interleukin-1 receptor antagonism on insulin and glucagon release of rat, mouse and human islets exposed to recombinant human interleukin-1 beta, and on interleukin-1 beta induced changes in blood glucose, serum insulin and serum glucagon levels in Wistar Kyoto rats. The interleukin-1 receptor antagonist reduced the co-mitogenic effect of interleukin-1 beta on mouse and rat thymocytes with a 50% inhibitory concentration of 10- and 100-fold molar excess, respectively. Complete inhibition was obtained with a 100-1,000-fold molar excess. However, at a 100-fold molar excess the interleukin-1 receptor antagonist did not antagonise the potentiating effect of interleukin-1 beta on rat islet insulin accumulation during 3 and 6 h of exposure or of interleukin-1 beta induced inhibition of insulin release after 24 h. In contrast, interleukin-1 beta stimulated islet glucagon release was completely antagonised by a 100-fold molar excess of interleukin-1 receptor antagonist. A 10,000-fold molar excess of interleukin-1 receptor antagonist was needed to antagonise interleukin-1 beta stimulatory and inhibitory effects on rat beta-cell function in vitro. A 100-fold excess of interleukin-1 receptor antagonist could not counteract interleukin-1 beta effects on mouse and human beta cells, excluding species difference in the efficacy of the human interleukin-1 receptor antagonist.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405745 TI - Intracellular and extracellular magnesium depletion in type 2 (non-insulin dependent) diabetes mellitus. AB - To investigate alterations of magnesium metabolism in Type 2 (non-insulin dependent) diabetes mellitus, we utilized a new magnesium-specific selective ion electrode apparatus to measure serum ionized magnesium (Mg-io) in fasting subjects with and without Type 2 diabetes, and compared these values to levels of serum total magnesium, and of intracellular free magnesium (Mgi) analysed by 31P NMR spectroscopy. Both Mg-io (0.630 +/- 0.008 vs 0.552 +/- 0.008 mmol/l, p < 0.001) and Mgi (223.3 +/- 8.3 vs 184 +/- 13.7 mmol/l, p < 0.001), but not serum total magnesium, were significantly reduced in Type 2 diabetes compared with non diabetic control subjects. Furthermore, a close relationship was observed between serum Mg-io and Mgi (r = 0.728, p < 0.001). We suggest that magnesium deficiency, both extracellular and intracellular, is a characteristic of chronic stable mild Type 2 diabetes, and as such, may predispose to the excess cardiovascular morbidity of the diabetic state. Furthermore, by more adequately reflecting cellular magnesium metabolism than total serum magnesium levels, Mg-io measurements may provide a more readily available tool than has heretofore been available to analyse magnesium metabolism in a variety of diseases. PMID- 8405746 TI - Partitioning the symptoms of hypoglycaemia using multi-sample confirmatory factor analysis. AB - The allocation of hypoglycaemic symptoms to autonomic or neuroglycopenic groups tends to occur on an a priori basis. In view of the practical need for clear symptom markers of hypoglycaemia more scientific approaches must be pursued. Substantial evidence is presented from two large scale studies we performed which support a three factor model of hypoglycaemic symptomatology, based on the statistical associations discovered among symptoms reported by diabetic patients. Study 1 involved 295 insulin-treated out-patients and found that 11 key hypoglycaemic symptoms segregated into three clear factors: autonomic (sweating, palpitation, shaking and hunger) neuroglycopenic (confusion, drowsiness, odd behaviour, speech difficulty and incoordination), and malaise (nausea and headache). The three factors were validated on a separate group of 303 insulin treated diabetic out-patients. Confirmatory factor analyses showed that the three factor model was the optimal model for explaining symptom covariance in each group. A multi-sample confirmatory factor analysis tested the rigorous assumptions that the relative loadings of symptoms on factors across groups were equal, and that the residual variance for each symptom was identical across groups. These assumptions were successful, indicating that the three factor model was replicated in detail across these two large samples. It is suggested that the results indicate valid groupings of symptoms that may be used in future research and in clinical practice. PMID- 8405747 TI - Quantification of human cytoplasmic islet-cell antibodies which cross-react with mouse pancreas: a follow-up study in type 1 (insulin-dependent) diabetic patients and in first-degree relatives. AB - We studied the heterogeneity of cytoplasmic islet-cell antibodies for cross reaction with mouse pancreas in 31 recent-onset Type 1 (insulin-dependent) diabetic patients and 31 first-degree relatives with islet-cell autoantibodies detected on human pancreas. Only six Type 1 diabetic patients displayed islet cell antibodies binding to human pancreas but not to mouse pancreas. Among 15 first-degree relatives displaying such antibodies which did not react with mouse pancreas, including one identical twin and one subject with polyglandular autoimmunity, none developed diabetes or even lost acute insulin response to intravenous glucose after 5 years of follow-up. By contrast, 14 of 20 (70%) of the Type 1 diabetic patients with islet-cell antibodies detected on human pancreas, and five first-degree relatives who progressed to a loss of acute insulin response to glucose and then to either Type 1 diabetes or glucose intolerance, also displayed antibodies reactive with mouse islets. Surprisingly, islet-cell antibodies were detectable on mouse pancreas but not on human pancreas in four Type 1 diabetic patients and in one relative who progressed to diabetes. In the five relatives who progressed to metabolic abnormalities, islet-cell antibody titres on mouse pancreas, quantified by the fluorescence intensity per islet at each serum dilution, progressively increased concomitantly with the loss of acute insulin response to glucose, whereas islet-cell antibody titres on human pancreas remained stable. The usefulness of such quantification was also validated by the fact that antibody titres on mouse pancreas were decreased after 3 months (p < 0.01) in recent-onset Type 1 diabetic patients, while titres on human pancreas were not.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405748 TI - Different effects of glucose and glyburide on insulin secretion in rat pancreatic islets pre-exposed to interleukin-1 beta. Possible involvement of K+ and Ca2+ channels. AB - In vitro islet exposure to interleukin 1 beta inhibits the beta-cell response to glucose. We have studied whether a similar inhibition also occurs in response to the sulphonylurea glyburide. Rat pancreatic islets were cultured for 24 h in the presence or absence of 50 U/ml interleukin 1 beta and then stimulated with either glucose or glyburide for 1 h at 37 degrees C. In control islets basal insulin secretion was 117 +/- 32 pg.islet-1.h-1 (mean +/- SEM, n = 7) and greatly increased in response to 16.7 mmol/l glucose (2140 +/- 293) or 10 mumol/l glyburide (1464 +/- 234). When islets were pre-exposed to interleukin 1 beta, insulin release was significantly reduced in response to glucose (323 +/- 80, p < 0.001) but not in response to glyburide (1316 +/- 185). Since both glucose and glyburide influence beta-cell K+ and Ca2+ efflux, to further investigate this different response in islets exposed to interleukin 1 beta we measured both Rb+ efflux (as index of the ATP-sensitive K+ channel activity) and Ca2+ uptake. In control islets, the increased insulin secretion in response to 16.7 mmol/l glucose or 10 mumol/l glyburide was associated with a reduction of 86Rb efflux (decrement of -50 +/- 1.2% and -49 +/- 2.3%, respectively, mean +/- SEM, n = 5). In contrast, in interleukin 1 beta pre-exposed islets both glucose and glyburide stimulation only slightly modified 86Rb efflux (decrement of -19 +/- 1.9% and 5.3 +/- 3.1%, respectively, n = 5, p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405749 TI - D-lysine reduces the non-enzymatic glycation of proteins in experimental diabetes mellitus in rats. AB - D-Lysine, the non-physiological isomer of L-lysine, can competitively reduce protein non-enzymatic glycation in vitro. To study the effect of D-lysine in vivo, 6-8-week old Sprague-Dawley rats with streptozotocin-induced diabetes mellitus were treated from diagnosis for 45 days with two daily subcutaneous injections of D-lysine (0.5 g.ml-1.day-1). Another group of diabetic rats was only injected with equal volumes of physiological saline (0.9% NaCl). Glycated haemoglobin was measured by ion exchange chromatography, and glycated serum and lens proteins by boronate affinity gel chromatography. Serum and urinary creatinine concentrations were evaluated by the alkaline-picrate reaction. Urinary lysine concentrations at mid- and end-study were evaluated by cation exchange chromatography. Blood glucose concentrations, serum creatinine levels and creatinine clearances, measured at the end of the study, were similar in both diabetic groups (> 22.0 mmol/l, < or = 106 mumol/l and approximately 0.02 ml/s, respectively). Urinary lysine concentration in D-lysine-treated diabetic animals was more than 50-fold higher than in placebo-treated diabetic rats. In D-lysine treated vs placebo-treated diabetic animals, a statistically significant reduction was found in the levels of glycated haemoglobin (stable HbA1; mean +/- SD = 3.00 +/- 0.74% vs 4.02 +/- 0.46%, p < 0.05; labile HbA1 = 3.92 +/- 0.89% vs 5.84 +/- 0.61%, p < 0.005), glycated serum proteins (1.40 +/- 0.47% vs 2.52 +/- 1.15%, p < 0.05) and glycated lens proteins (4.90 +/- 0.96% vs 5.98 +/- 0.65%, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405750 TI - Inhibition of matrix-induced bone differentiation by advanced glycation end products in rats. AB - Glycation of long-lived proteins is an inevitable consequence of aging that is accelerated in patients with diabetes mellitus. Treatment of demineralized bone matrix particles from 35-week-old normal Long-Evans rats with glycoaldehyde, a precursor of advanced glycation end-products, was used to assess the effects of bone-matrix glycation on the process of bone differentiation. Matrix was incubated in phosphate buffered saline alone, phosphate buffered saline containing glycolaldehyde, glycolaldehyde plus the advanced glycation product inhibitor aminoguanidine, or glycolaldehyde plus the advanced glycation product inhibitor sodium cyanoborohydride. Glycolaldehyde increased the matrix advanced glycation product content as measured by specific fluorescence more than two fold, while inhibiting bone differentiation more than 90% as measured by in vivo 45CaCl2 uptake, alkaline phosphatase levels, and histology. In contrast, simultaneous incubation with the advanced glycation product-inhibitor aminoguanidine or sodium cyanoborohydride not only reduced fluorescence to normal, but also restored bone differentiation. Furthermore, the inhibition of bone differentiation by glycolaldehyde was not reversed by subsequent application of recombinant bone morphogenetic protein-2. These observations suggest that formation of advanced glycation products on bone matrix alters its ability to induce bone formation, and probably involves alterations of binding sites for extractable proteins with direct bone inductive properties such as bone morphogenetic protein-2. Decreased bone formation associated with aging and diabetes may result, in part, from advanced glycation product formation on matrix proteins. PMID- 8405751 TI - Regulation of glucose transporter (GLUT 3) and aldose reductase mRNA inbovine retinal endothelial cells and retinal pericytes in high glucose and high galactose culture. AB - The regulation of GLUT-3 and aldose reductase mRNA in retinal endothelial cells and retinal pericytes was studied in response to variations in the extracellular concentration of hexoses. In physiological concentrations of glucose (5 mmol/l), an increase in the level of GLUT-3 mRNA was observed in cultured cells compared to the level of mRNA found in the absence of glucose. In contrast, there was little change in the level of GLUT-3 mRNA when the cells were cultured in the presence of 5 mmol/l galactose. In high concentrations of glucose, there was a decline in GLUT-3 mRNA indicating that the GLUT-3 mRNA is regulated by the extracellular concentration of glucose. In contrast, at both 5 mmol/l and 25 mmol/l glucose, the level of aldose reductase mRNA was increased. Furthermore, there were differences in the magnitude of the increase of aldose reductase mRNA between bovine retinal pericytes and bovine retinal endothelial cells with a greater increase being observed in the pericytes. We propose that this demonstration of a facilitative glucose transporter system within retinal cells, and in particular the specific response to different hexoses and the known distinct kinetic parameters of the transporter system in specific cell types, highlights the heterogeneity of hexose transport mechanisms in retinal cells. Thus, hypergalactosaemia as a model system for the study of diabetic retinopathy should be used with caution. PMID- 8405752 TI - Can insulin administration cause an acute metabolic acidosis in vivo? An experimental study in dogs. AB - Insulin is the cornerstone of therapy for diabetic ketoacidosis because it causes the rate of ketoacid production to fall; this action takes several hours to occur. Insulin also causes H+ to be transported from the intracellular fluid to the extracellular fluid in vitro. The purpose of this study was to determine if insulin led to the acute export of H+ from the intracellular fluid in vivo. If so, we wished to determine if this also occurred during chronic metabolic acidosis, to quantitate the magnitude of the H+ shift, and to evaluate the mechanisms involved. The administration of low- or high-dose insulin to normal dogs and high-dose insulin to dogs with chronic metabolic acidosis caused the concentration of bicarbonate in plasma to decline by close to 3 mmol/l. The PCO2 fell by close to 15% in all three groups of dogs, so one component of the fall was due to hyperventilation. As the pH of blood did not change, a primary metabolic acidosis also occurred. The fall in bicarbonataemia was not due to net accumulation of organic acids or to a loss of bicarbonate or organic anions in the urine. Taken together, insulin, when given at doses used to treat diabetic ketoacidosis, might induce a significantly greater degree of acidaemia in the extracellular fluid acutely after it is given. PMID- 8405753 TI - Reversible impairment of glucose tolerance in patients with endemic fluorosis. Fluoride Collaborative Study Group. AB - Endemic fluorosis is a condition resulting from prolonged ingestion of drinking water which contains excess fluoride. Studies on rats have suggested that fluoride toxicity may produce glucose intolerance and abnormalities in insulin secretion. We studied glucose and insulin profiles following an oral glucose load in patients with endemic fluorosis. Twenty-five young adults (age range, 15-30 years) with endemic fluorosis, and an equal number of matched healthy control subjects with normal fluoride intake were studied. Impaired glucose tolerance was demonstrated in 10 of 25 (40%) patients with endemic fluorosis. Patients with impaired glucose tolerance had significantly higher fasting serum immunoreactive insulin (p < 0.05), higher fasting serum fluoride (p < 0.001), and a significantly lower fasting glucose to insulin ratio than that in patients with normal glucose tolerance (p < 0.001) or control subjects (p < 0.05). The fasting serum fluoride levels correlated positively with the area under the glucose curve (r = 0.80, p < 0.01) in patients with impaired glucose tolerance. Interestingly these abnormalities could be reversed when the village was provided drinking water with fluoride levels within acceptable limits. The present study shows that chronic fluoride toxicity in humans could result in significant abnormalities in glucose tolerance which are reversible upon removal of the excess fluoride. PMID- 8405754 TI - Effect of protein intake on glycaemic control and renal function in type 2 (non insulin-dependent) diabetes mellitus. AB - Recent clinical investigations have suggested that dietary protein intake may modulate the progression of diabetic nephropathy and influence glycaemic control in Type 2 (non-insulin-dependent) diabetes mellitus. Twelve normotensive Type 2 diabetic patients with microalbuminuria took part in a randomized cross-over trial of a 3-week high protein diet (2.0 g/kg.desirable weight per day) and a 3 week moderate protein diet (0.8 g/kg desirable weight per day) to test the simultaneous effect of protein intake modulation on glycaemic control and renal function. Both diets were isoenergetic and the moderate protein diet was supplemented with calcium and phosphate. Renal function and glycaemic control were evaluated at the beginning and at the end of each diet. The moderate protein diet reduced the urinary albumin excretion rate, glomerular filtration rate, creatinine clearance, and proteinuria without adversely affecting glycaemic control; fasting glycaemia and the ratio of fructosamine to proteins were significantly reduced. The high protein diet induced similar improvements in glycaemic control but small changes in renal function. PMID- 8405755 TI - UK Prospective Diabetes Study (UKPDS). IX: Relationships of urinary albumin and N acetylglucosaminidase to glycaemia and hypertension at diagnosis of type 2 (non insulin-dependent) diabetes mellitus and after 3 months diet therapy. AB - In 672 newly-diagnosed, Type 2 (non-insulin-dependent) diabetic patients without urinary infection, aged 51 (9) years, mean (1 SD), 28% of patients had raised albuminuria, defined as albumin excretion greater than 25 mg/l and 66% raised urinary N-acetylglucosaminidase excretion defined as greater than 300 mumol.h-1.l 1 (both urinary analytes corrected by linear regression on urinary creatinine to 10 mmol/1). In a univariate analysis, urinary N-acetylglucosaminidase and albumin excretion correlated with each other (rs = 0.42, p < 0.001), and with fasting plasma glucose (rs = 0.46 and rs = 0.27, p < 0.001, respectively). The association of urinary albumin and N-acetylglucosaminidase remained significant after taking the fasting plasma glucose levels into account, partial rs = 0.34, p < 0.001. After 3 months of dietary therapy BMI decreased from 29.7 (5.9) kg/m2 to 28.8 (5.8) kg/m2, fasting plasma glucose levels from 12.2 (3.8) mmol/l to 9.8 (3.8) mmol/l, and systolic blood pressure from 143 (21.8) mmHg to 131 (20.3) mmHg, p < 0.001 for each variable. There were concomitant decreases in urinary N acetylglucosaminidase, geometric mean (1 SD interval), 397 mumol.h-1.l-1 (216 to 728) to 291 mumol.h-1.l-1 (160 to 528), p < 0.001 and in albumin excretion 16 mg/l (5 to 51) to 13 mg/l (4 to 40), p < 0.001. The decrease in urinary N acetylglucosaminidase, but not the decrease in urinary albumin excretion, was associated with the initial degree of glycaemia and the decrease in glycaemia in response to diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405756 TI - Inhibition of insulin secretion, but normal peripheral insulin sensitivity, in a patient with a malignant endocrine pancreatic tumour producing high amounts of an islet amyloid polypeptide-like molecule. AB - Islet amyloid polypeptide or amylin is a polypeptide secreted mainly from the pancreatic beta cells together with insulin upon stimulation. High levels of islet amyloid polypeptide have also been shown to increase the peripheral insulin resistance and consequently a role for islet amyloid polypeptide in the glucose homeostasis has been suggested. We have studied the glucose homeostasis in a patient with a malignant endocrine pancreatic tumour producing large amounts of an islet amyloid polypeptide-like molecule (about 400 times the upper reference level for islet amyloid polypeptide). This patient developed insulin-requiring diabetes mellitus shortly after the tumour diagnosis. Both intravenous and oral glucose tolerance tests revealed inhibited early responses in insulin and C peptide release, but the insulin and C-peptide response to glucagon stimulation was less affected. Aneuglycaemic insulin clamp showed normal insulin-mediated glucose disposal. In vitro experiments, where isolated rat pancreatic islets were cultured with serum from the patient, showed a moderately decreased islet glucose oxidation rate and glucose-stimulated insulin release compared to islets cultured with serum from healthy subjects. However, culture of rat islets with normal human serum supplemented with synthetic rat islet amyloid polypeptide did not affect the glucose-stimulated insulin release. In conclusion, the observed effects show that the diabetic state in this patient was associated with an impaired glucose-stimulated insulin release but not with an increased peripheral insulin resistance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405757 TI - Cell membrane fatty acid composition in type 1 (insulin-dependent) diabetic patients: relationship with sodium transport abnormalities and metabolic control. AB - We have studied the fatty acid composition of erythrocyte membrane phospholipids in nine Type 1 (insulin-dependent) diabetic patients and nine healthy control subjects. Cell membranes from the diabetic patients showed a marked decrease in the total amount of polyunsaturated fatty acids (19.0% +/- 2.2 vs 24.6% +/- 1.4, p < 0.0001) mainly at the expense of docosahexaenoic acid C22:6(n3) (2.9% +/- 1.1 vs 5.3% +/- 1.3, p < 0.001), and arachidonic acid C20:4n6 (12.0% +/- 1.6 vs 15.1% +/- 0.6, p < 0.0005). Conversely, the total amount of saturated fatty acids was significantly increased (p < 0.05) and the polyunsaturated/saturated ratio was decreased in the Type 1 diabetic patients (p < 0.00 005). Neither the time from diagnosis, nor C-peptide levels, correlated with parameters indicating a poor metabolic control of Type 1 diabetes. However, C22:6(n-3) and total n-3 content significantly correlated with HbA1c (r = -0.79 and r = -0.88, respectively, p < 0.01), fructosamine (r = -0.71 and r = -0.74, respectively, p < 0.05), and Na+-K+ ATPase activity (maximal rate/Km quotient) (r = 0.78 and r = 0.71, respectively, p < 0.05). In conclusion we have found marked alterations of cell membrane lipid composition in Type 1 diabetic patients. These cell membrane abnormalities in lipid content were related to sodium transport systems and to poor metabolic control. Either diet, or the diabetic state, might be responsible for the observed cell membrane abnormalities. A dietary intervention study might differentiate the role of diet and diabetes in the reported cell membrane alterations. PMID- 8405758 TI - Relationship between oral glucose tolerance and gastric emptying in normal healthy subjects. AB - The relationships between gastric emptying and intragastric distribution of glucose and oral glucose tolerance were evaluated in 16 healthy volunteers. While sitting in front of a gamma camera the subjects drank 350 ml water containing 75 g glucose and 20 MBq 99mTc-sulphur colloid. Venous blood samples for measurement of plasma glucose, insulin and gastric inhibitory polypeptide were obtained at- 2, 2,5,10,15,30,45,60,75,90,105,120 and 150 min. Gastric emptying approximated a linear pattern after a short lag phase (3.3 +/- 0.8 min). The 50% emptying time was inversely related to the proximal stomach 50% emptying time (r = -0.55, p < 0.05) and directly related to the retention in the distal stomach at 120 min (r = 0.72, p < 0.01). Peak plasma glucose was related to the amount emptied at 5 min (r = 0.58, p < 0.05) and the area under the blood glucose curve between 0 and 30 min was related to the amount emptied at 30 min (r = 0.58, p < 0.05). In contrast, plasma glucose at 120 min was inversely related to gastric emptying (r = -0.56, p < 0.05) and plasma insulin at 30 min (r = -0.53, p < 0.05). Plasma insulin at 120 min was inversely related (r = -0.65, p < 0.01) to gastric emptying. The increase in plasma gastric inhibitory polypeptide at 5 min was related directly to gastric emptying (r = 0.53, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405759 TI - The association of physical activity with obesity, fat distribution and glucose intolerance in Pima Indians. AB - The relationships between physical activity, obesity, fat distribution and glucose tolerance were examined in the Pima Indians who have the highest documented incidence of non-insulin-dependent diabetes. Fasting and 2-h post-load plasma glucose concentrations, body mass index, and waist-to-thigh circumference ratios were determined in 1054 subjects aged 15-59 years. Current (during the most recent calendar year) and historical (over a lifetime) leisure and occupational physical activity were determined by questionnaire. Current physical activity was inversely correlated with fasting and 2-h plasma glucose concentrations, body mass index and waist-to-thigh ratios for most sex-age groups even when diabetic subjects were excluded. Controlled for age, obesity and fat distribution, activity remained significantly associated with 2-h plasma glucose concentrations in males. In subjects aged 37-59 years, individuals with diabetes compared to those without reported significantly less leisure physical activity during the teenage years (median hours per week of activity, 9.1 vs 13.2 for men; 1.0 vs 2.2 for women). Controlled for body mass index, sex, age and waist-to thigh ratio, subjects who reported low levels of historical leisure physical activity had a higher rate of diabetes than those who were more active. In conclusion, current physical activity was inversely related to glucose intolerance, obesity and central distribution of fat, particularly in males. Subjects with diabetes were currently less active and reported less historical physical activity than non-diabetic subjects. These findings suggest that activity may protect against the development of non-insulin-dependent diabetes both directly and through an influence on obesity and fat distribution. PMID- 8405760 TI - A nationwide population-based study of the familial aggregation of type 1 (insulin-dependent) diabetes mellitus in Denmark. Danish Study Group of Diabetes in Childhood. AB - The objective of the present study was to assess the prevalence of familial aggregation of Type 1 (insulin-dependent) diabetes mellitus among Danish families with a diabetic child aged 20 years or less and to compare epidemiological data for familial and sporadic cases. We attempted to identify all patients with Type 1 diabetes aged 0-19 years in Denmark treated at paediatric departments or at departments of internal medicine. This comprises more than 98% of all patients with Type 1 diabetes in this age group. Patients were identified through the local diabetic out-patient registry and asked to complete a questionnaire regarding data on diabetes onset and family history. Of 1574 probands 1419 agreed to participate (90.2%). Additional cases of Type 1 diabetes were found in 171 families (12.8%). Of these 115 were parent-offspring affected families, and in 56 families at least two siblings had Type 1 diabetes and healthy parents. Significant correlation in age at onset of Type 1 diabetes in concordant siblings was observed (r = 0.5, p = 0.0004). Significantly more probands had an affected father with Type 1 diabetes than a mother affected (p < 0.0001). Heterogeneity in epidemiological characteristics was observed between familial and sporadic cases, i.e. familial index cases were younger at onset of the disease, their parents were younger at birth of the index case, and there was no difference in gender of familial cases in contrast to sporadic cases where significantly more males were found. Over a 4-year period (1986-1989) an increasing trend in incidence was observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405761 TI - Insulin resistance syndrome: possible key role of blood flow in resting muscle. PMID- 8405762 TI - Insulin resistance and mechanical dysfunction in hearts of Wistar rats with streptozotocin-induced non-insulin-dependent diabetes mellitus. PMID- 8405763 TI - Plasma lipoprotein (a) concentration in diabetes mellitus. PMID- 8405764 TI - Two distinct adhesion systems are responsible for EDTA-sensitive adhesion in Dictyostelium discoideum. AB - Early in their developmental program, Dictyostelium discoideum exhibit EDTA sensitive and EDTA-resistant adhesion. The molecules which mediate the adhesions have been called contact sites, with contact sites A mediating EDTA-resistant adhesion and contact sites B mediating EDTA-sensitive adhesion. The studies described here have revealed that prior to aggregation, a second EDTA-sensitive adhesion system emerges. In keeping with previously established nomenclature, the molecules mediating the newly discovered adhesion system have been called contact sites C. Unlike contact sites B, contact sites C are unaffected by a contact sites B-blocking peptide. Contact sites C-mediated adhesion is also distinct from contact sites B-mediated adhesion in that contact sites C-mediated adhesion is EGTA-resistant and in the presence of EDTA it can be rescued by the addition of Mg2+. Thus Mg2+ may be the cation present under physiological conditions that is essential for contact sites C activity. Unlike contact sites B-mediated adhesion, contact sites C-mediated adhesion is not observed in growing amoebae. Contact sites C-mediated adhesion first becomes apparent within hours after the initiation of development and its strength appears to increase throughout the first 10 h of the developmental program. A mutant lacking the EDTA-resistant contact sites A exhibits normal contact sites B- and C-mediated adhesion, demonstrating that both EDTA-sensitive adhesion systems are independent of contact sites A. Thus aggregating D. discoideum amoebae possess three distinct adhesion systems, one of them is EDTA-resistant and the other two are EDTA sensitive. PMID- 8405765 TI - Estrogen- and temperature-induced medullary cord regression during gonadal differentiation in a turtle. AB - Gonadal differentiation associated with estrogen-induced female sex determination was examined in a turtle with temperature-dependent sex determination, and was compared to ovarian differentiation at a female-producing temperature. Freshly ladi eggs of the red-eared slider, Trachemys scripta, were incubated at a male producing temperature (26 degrees C) and were experimentally manipulated at one of three embryonic stages: stage 15, 17, or 20 (i.e. early, midway, or late in the temperature-sensitive and estrogen-sensitive periods). At those developmental stages, groups of eggs were either: (1) treated with a control solution (95% ethanol) and placed back at the male-producing temperature, (2) treated with 10 micrograms of estradiol-17 beta and placed back at the male-producing temperature, or (3) shifted to a female-producing temperature (31 degrees C). Additionally, a control group of freshly laid eggs was continually incubated at 31 degrees C throughout embryonic development. To examine morphological events occurring after the treatments, a subset of embryos from each group was examined at the time of the treatment and at 1-2 stage intervals following the treatments. The results indicate that estradiol-17 beta as well as female-producing temperature may ensure female sex determination by facilitating medullary cord regression. Further, the results reveal a chronology of differentiation in which medullary cord regression temporally precedes cortical proliferation. PMID- 8405766 TI - Activity of protein kinase C during the differentiation of chick limb bud mesenchymal cells. AB - To investigate the relationship between protein kinase C (PKC) and chondrogenesis, PKC activity was assayed in cultures of stage 23/24 chick limb bud mesenchymal cells under various conditions. PKC activities of cytosolic and particulate fractions were low in 1 day cultured cells. As chondrogenesis proceeds, cytosolic PKC activity increased more than twofold, while that of the particulate fraction increased only slightly. Three days' treatment of cultures with phorbol-12-myristate-13-acetate (PMA, 5 x 10(-8) M) inhibited chondrogenesis judged by the accumulation of Alcian blue bound to the extracellular matrix and depressed PKC activity in cytosolic fraction. When cells were grown for 3 days in control medium after 3 days' treatment with PMA, chondrogenesis resumed and PKC activity recovered to normal values. PKC activity in cultures plated at low density (5 x 10(6) cells/ml) where chondrogenesis is reduced was as low as that in 1 day cultured cells plated at high density (2 x 10(7) cells/ml) or that in PMA treated cells. On the other hand, staurosporine promoted chondrogenesis without affecting PKC activity. Furthermore, reversal of PMA's inhibitory effect on chondrogenesis by staurosporine was not accompanied by recovery of PKC activity. These data indicate that increases in PKC activity is closely related to chondrogenesis and that PMA inhibits chondrogenesis by depressing PKC. However, staurosporine's enhancing effect on chondrogenesis is not related to PKC activity. PMID- 8405767 TI - Pancreatic morphogenesis and extracellular matrix organization during rat development. AB - We investigated the rat pancreatic morphology at various developmental stages ranging from 12 days of gestation to the neonatal stage, with special emphasis on alterations in extracellular matrix organization in vivo. The rat pancreatic development in utero could be divided into four representative stages as follows: (1) initial epithelial buds (12 days of gestation), (2) elongated and branching epithelium (13-14 days), (3) tubular structure (15-16 days), and (4) acinar structure (17 days or more). Ultrastructurally, the fetal and neonatal pancreata were almost constantly encompassed by continuous basal lamina, except for the earliest stage, in which minute disruptions of basal lamina were observed. Through the disruption, the direct epithelial-mesenchymal contact was formed between an endocrine cell and an adjacent mesenchymal cell, which implied epithelial-mesenchymal interactions in processes of endocrine cell differentiation. Collagen fibrils were frequently accumulated at the cleft (branchpoint) of the branching epithelium during the second and third stages mentioned above. Immunohistochemically, fibronectin and collagen type-I were localized particularly beside the neck (narrow part) or cleft of the pancreatic epithelium at these stages, although continuous linear localization of these matrices was noted around the initial pancreatic bud. This was in contrast to invariable linear localization of laminin and collagen type-IV at the epithelial/mesenchymal interface throughout the pancreatic development. Diffuse fibrillar localization of fibronectin and collagen type-I in the mesenchyme was pronounced at the later stages and after birth. Collagen type-III was only focally detectable around the pancreatic epithelium from the second stage, and its distinct localization was noted in the interlobular connective tissue after birth. Thus, chronological changes in extracellular matrix organization seemed to be closely related to morphogenetic processes of the rat pancreas, especially in the branching epithelial morphogenesis, and the major alterations appeared prior to distinct acinar cell differentiation. PMID- 8405768 TI - Differences in the effects of treatment of uncompacted and compacted mouse embryos with phorbol esters on pre- and postimplantation development. AB - Differences are described in the effects of treatment of preimplantation mouse embryos with low levels (0.01-1 nM) of phorbol myristate acetate (PMA), during three different periods of a 48-h culture from the 2-cell stage, on pre- and postimplantation development. Treatment of embryos with PMA for 48 h (first group) or 24 h (second group) from the 2-cell stage caused premature cavitation (prior to the 16-cell stage) and it also reduced the size and alkaline phosphatase (ALPase) activity of inner cell masses (ICMs), as well as the numbers of cells in blastocysts, in a dose-dependent manner. Treatment of early morulae with PMA for 24 h (third group) did not have the above mentioned effects on embryos but inhibited the formation and subsequent enlargement of the blastocoel. The blastocysts that were allowed to develop in the three treatment groups were examined for postimplantation development. Implantation was unaffected in all groups. The survival rate after implantation was low in the first and second groups but relatively high in the third group. The results indicate that an embryo exposed to PMA for 24 h from the 2-cell stage forms a premature blastocoel, and, in such an embryo, quantitative and qualitative differentiation into the ICM is blocked but qualitative differentiation into trophectoderm is uninhibited. Consequently, the embryo can implant but does not survive for a long time. When embryos were exposed to PMA for 24 h from the early morula stage, the formation and enlargement of the blastocoel were inhibited even though the treatment had a minimal effect on other developmental events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405769 TI - Kinetics of in vitro mineralization by an osteogenic clonal cell line (C1) derived from mouse teratocarcinoma. AB - We have previously reported the isolation of an osteogenic clonal cell line (C1) derived from mouse teratocarcinoma and immortalized by the SV 40 oncogenes. In this report we describe the kinetics of osteogenic differentiation of aggregated C1 cells by following the matrix deposition and mineralization and the expression of alkaline phosphatase. We show that after addition of beta-glycerophosphate and ascorbic acid, more than 95% of C1 aggregates synthesize a bone matrix which is deposited as early as 2 days and increases progressively with time in culture. Matrix calcification is evidenced by von Kossa staining and tetracycline incorporation into the mineral whereas no calcification appears in control cultures. Calcium is detectable in mineralizing aggregates at 2 days and calcium content increases linearly with time in culture, being 125-fold higher in mineralizing nodules than in control aggregates at 30 days. Aggregated C1 cells are characterized by a high activity of the bone type isoenzyme of alkaline phosphatase, a marker of osteoblast phenotype. Upon addition of inducers, alkaline phosphatase activity decreases by five-fold after the onset of mineralization and remains stable thereafter. The down-regulation of alkaline phosphatase activity is confirmed at the cellular level by histochemical staining. The mRNA levels for alkaline phosphatase decline during osteogenesis, following a pattern similar to the decrease in protein activity. Analysis of DNA synthesis by (3H)-thymidine incorporation and quantification of labelled nuclei on autoradiographs shows that C1 cells proliferation is not down-regulated during the time course of differentiation and that proliferating C1 cells still express alkaline phosphatase activity during osteogenic differentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405770 TI - Tissue-specific and differentiation-appropriate expression of the human involucrin gene in transgenic mice: an abnormal epidermal phenotype. AB - Involucrin is a precursor of the keratinocyte cornified envelope that is specifically expressed in the suprabasal layers of the epidermis and other stratifying squamous epithelia. To study involucrin gene expression and the function of involucrin, we expressed a 6 kb DNA fragment of the human involucrin gene, containing approximately 2.5 kb of upstream sequence and 0.5 kb of downstream sequence, in transgenic mice. The transgene produces a 68 kDa protein that is detected by a human involucrin-specific antibody, and is expressed in a tissue-specific and differentiation-appropriate manner (i.e., expression is confined to the suprabasal layers of the epidermis, extocervix, trachea, esophagus and conjunctiva). Soluble involucrin levels are two to four times higher in transgenic epidermal keratinocytes compared to human foreskin keratinocytes. Newborn heterozygous animals have a normal birth weight and a normal appearing epidermis and hair growth begins at 4 to 5 days of age (i.e., the same time as hair growth in non-transgenic animals). In a subpopulation of the newborn homozygous animals birth weight is reduced, the epidermis is scaly and hair growth begins late, at around 9 to 10 days of age. In addition, the hair tends to stand erect on both heterozygous and homozygous adult animals giving the appearance of diffuse alopecia. Immunofluorescent and electron microscopy localize involucrin in the hair follicle and cornified envelope, respectively. These results suggest that overexpression of involucrin may cause abnormalities in hair follicle structure/function and cornified envelope structure. These animals provide a new model for the study of cornified envelope structure and function. PMID- 8405771 TI - Matrix metalloproteases of the developing sea urchin embryo. AB - A distinct group of metalloproteases has been identified in the developing sea urchin embryo by gelatin substrate gel zymography, a highly sensitive protease detection assay. The developing Arbacia embryo exhibited four prominent bands of gelatinase activity with apparent molecular masses of 55, 50, 42 and 38 kDa. The activity of the 55, 42 and 38 kDa tissue gelatinases increased and that of the 50 kDa tissue gelatinase decreased during embryonic development. All four enzymes were EDTA- and 1,10-phenanthroline sensitive and phenyl methyl sulphonyl fluoride (PMSF) insensitive. None of the enzymes had detectable caseinolytic activity in casein substrate gels. Although the Arbacia enzymes possessed a number of properties that are characteristic of the mammalian matrix metalloprotease family, they did not appear to be converted to lower molecular weight forms by organomercurial treatment and are distinct in this aspect. The Arbacia metalloproteases are candidate enzymes for the tissue and matrix remodeling that occurs during sea urchin embryo development. PMID- 8405772 TI - Expression patterns of loricrin in various species and tissues. AB - In this study we analyzed the expression patterns of loricrin in various species and tissues using immunohistochemistry, immunoblotting and Northern blots. Loricrin is a glycine-, serine- and cysteine-rich protein expressed very late in epidermal differentiation in the granular layers of normal mouse and human epidermis. Later on in differentiation, loricrin becomes crosslinked as a major component into the cornified cell envelope by the formation of N epsilon-(gamma glutamyl)lysine isopeptide bonds. This process either occurs directly or by the intermediate accumulation in L-keratohyaline granules of mouse epidermis and human acrosyringia. Loricrin was identified in all mammalian species analyzed by virtue of its highly conserved carboxy-terminal sequences revealing an electric mobility of approximately 60 kDa in rodents, rabbit and cow and of approximately 35 kDa in lamb and human on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Loricrin is expressed in the granular layer of all mammalian orthokeratinizing epithelia tested including oral, esophageal and fore-stomach mucosa of rodents, tracheal squamous metaplasia of vitamin A deficient hamster and estrogen induced squamous vaginal epithelium of ovary ectomized rats. Loricrin is also expressed in a few parakeratinizing epithelia such as BBN [N butyl-N-(4-hydroxybutyl)nitrosamine]-induced murine bladder carcinoma and a restricted subset of oral and single vaginal epithelial cells in higher mammals. Our results provide further evidence that the program of squamous differentiation in internal epithelia of the upper alimentary tract in rodents and higher mammals differ remarkably. In addition, we also have noted the distinct distribution patterns of human loricrin and involucrin, another major precursor protein of the cornified cell envelope. PMID- 8405773 TI - Proliferation and differentiation of human fetal myoblasts is regulated by PDGF BB. AB - A myoblast clone, G6, was obtained from thigh muscle of an 11 week old human fetus, and used to examine the effect of platelet-derived growth factor (PDGF) on cell multiplication and differentiation. G6 myoblasts showed extensive fusion, and expressed creatine phosphokinase activity and muscle specific gene mRNA (myosin heavy chain, alpha-actin) when switched to a differentiation medium. The cells expressed PDGF beta-receptor mRNA, and bound 125I-PDGF-BB specifically. Expression of PDGF beta-receptors declined during in vitro differentiation. Relative levels of transcripts for the myogenic regulatory factors Myf4 (myogenin), Myf5, and Myf6 (MRF4) increased during the differentiation process, whereas Myf3 (MyoD1) was preferentially expressed in undifferentiated myoblasts. Treatment of the myoblasts with PDGF-BB increased DNA synthesis and cell density. Myogenic differentiation, analyzed as number of nuclei present in myotubes and expression of creatine phosphokinase and myosin heavy chain, was partly inhibited by the presence of PDGF-BB in the differentiation medium. PDGF-BB may, therefore, have the potential of regulating human muscle development and muscle regeneration. PMID- 8405774 TI - Subtypes of non-transformed human mammary epithelial cells cultured in vitro: histo-blood group antigen H type 2 defines basal cell-derived cells. AB - Normal (non-transformed) human mammary epithelial cell lines derived from reduction mammoplasties were analyzed by immunocytochemistry with more than 80 monoclonal antibodies (mAbs) and other specific reagents to tissue-specific and developmentally regulated antigens at different passage levels. A subpopulation of poorly differentiated, proliferating epithelial cells, corresponding to the 'selected' cell type of late passages, is shown to be characterized by a new marker, the histo-blood group antigen H type 2, probably carried on a membrane bound glycolipid. These cells also express a number of other onco-developmental carbohydrate antigens [Le(y), Le(x), sialosyl-Le(a), precursor of Thomsen Friedenreich antigen (Tn), but not Thomsen-Friedenreich antigen and sialosyl-Tn]. Their cytokeratin (CK) phenotype, as assessed by reactivity with monospecific mAbs and two-dimensional gel electrophoresis, is CK 5, 6, 14 and 17, with CK 19 being consistently absent, and varying minor amounts of CK 7, 8 and 18, as well as 15 and 16. The reactivity of these cells with a panel of 11 mAbs specific for CK 18 varies considerably even after cloning, indicating heterogeneity of epitope expression or accessibility. Our data strongly suggest that the H type 2+ cells develop from the basal cell layer of the mammary gland. PMID- 8405775 TI - Thirteen-week, repeated inhalation exposure of F344/N rats and B6C3F1 mice to ferrocene. AB - Ferrocene (dicyclopentadienyl iron; CAS No. 102-54-5) is a relatively volatile compound used as a chemical intermediate, a catalyst, and an antiknock additive in gasoline. This organometallic chemical is of particular interest because of its structural similarities to other metallocenes, some of which are carcinogenic. F344/N rats and B6C3F1 mice were exposed to 0, 3.0, 10, and 30 mg ferrocene vapor/m3, 6 hr/day, 5 days/week, for 13 weeks. During these exposures, no rats or mice died, nor were any clinical signs of ferrocene-related toxicity observed. At the end of the exposures, male rats exposed to the lowest and highest level of ferrocene had decreased body weight gains compared to filtered air-exposed control male rats, while body weight gains for all groups of both ferrocene- and filtered-air-exposed female rats were similar. Male mice exposed to ferrocene had no differences in body weight gains, compared to controls, but female mice had decreases in body weight gains at the 10 and 30 mg/m3 exposure levels. There were exposure concentration- and exposure-time-related increases in lung burdens of iron. The mean iron lung burden in rats exposed to 30 mg ferrocene vapor/m3 for 90 days was four times greater than the burden in control rats. No exposure-related changes in respiratory function, lung biochemistry, bronchoalveolar lavage cytology, total lung collagen, clinical chemistry, and hematology parameters were observed. This suggests that the accumulations of iron in lung did not cause an inflammatory response nor any functional impairment of the lung. There were no indications of developing pulmonary fibrosis nor of any hematologic toxicity. No exposure-related gross lesions were seen in any of the rats or mice at necropsy. Exposure-related histopathologic alterations, primarily pigment accumulations, were observed in the nose, larynx, trachea, lung, and liver of both species, and in the kidneys of mice. Lesions were most severe in the nasal olfactory epithelium where pigment accumulation, necrotizing inflammation, metaplasia, and epithelial regeneration occurred. Nasal lesions were observed in all ferrocene-exposed animals and differed only in severity, which was dependent on the exposure concentration. Histochemical stains of these target tissues showed the presence of iron ions. The results suggest that the mechanism of ferrocene toxicity may be the intracellular release of ferrous ion through ferrocene metabolism, followed by either iron-catalyzed lipid peroxidation of cellular membranes or the iron-catalyzed Fenton reaction to form hydroxyl radicals that directly react with other key cellular components, such as protein or DNA. PMID- 8405776 TI - Effects of 1,1-dichloroethene and of some of its metabolites on the functional viability of mouse hepatocytes. AB - 1,1-Dichloroethene (DCE) is hepatotoxic in rodents, and the expression of its toxicity involves probably its metabolism. In this study the role of DCE metabolites in the generation of the hepatotoxic lesion was investigated. Hepatocytes from male BALB/c mice in suspension were used as the experimental model. Cells were incubated with DCE for up to 5 hr and cellular viability was assessed by measurement of the release of lactate dehydrogenase into the medium and by alterations in the reduction of the dye 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide. After incubation for 3 hr DCE at 0.5 mM caused maximal toxicity, whereas at 0.1 mM DCE was only marginally toxic. Cytotoxicity was exacerbated by pretreatment of mice with buthionine sulfoximine (1.6 g/kg), an inhibitor of glutathione biosynthesis, given 4 hr prior to hepatocyte isolation. Inclusion of N-acetylcysteine (10 mM) into the incubate protected cells against DCE-induced cytotoxicity. Coincubation with octylamine (0.5 mM), an inhibitor of cytochrome P450, abolished the cytotoxic potential of 0.5 mM DCE during incubation for 3 hr. DCE toxicity was increased in hepatocytes from mice which had received ethanol or acetone in their drinking water, both of which induce levels of the hepatic cytochrome P450 isozyme P450 2E1. Incubation of cells with the P450 2E1 inhibitors N,N-dimethylformamide (10 mM) or diethyldithiocarbamate (100 microM) protected liver cells against the detrimental effect of DCE. Pretreatment of animals with phenobarbital, which induces the P450 2B subfamily, or 3-methylcholanthrene, which induces P450 1A1, did not affect the degree of hepatocytotoxicity elicited DCE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405777 TI - Hepatocyte spheroids: prolonged hepatocyte viability for in vitro modeling of nongenotoxic carcinogenesis. AB - To explore peroxisome proliferator-perturbed hepatocyte growth regulation, robust in vitro models of liver are required. This has always posed a problem since isolated hepatocytes show a rapid loss of viability and differentiation status and cease to be useful after 3-4 days in culture. We now describe a model system in which rat hepatocytes are maintained as three-dimensional spheroids. The maintenance of hepatocyte viability and morphology in these cultures is considerably prolonged over that seen in monolayer culture and is comparable to that obtained by the use of collagen gels or dimethyl sulfoxide. The spheroid system is, however, free of any additives that may lead to artifact and free of excessive exogenous protein that may compromise subsequent analyses. Ultrastructural examination reveals extensive interhepatocyte junctional complexes and interdigitation of adjacent membranes together with the presence of bile cannalicular structures. Furthermore, hepatocytes maintained as spheroids retain expression of liver markers such as albumin and also retain their ability to respond to peroxisome proliferators: even after 12 days in culture, treatment with the peroxisome proliferator nafenopin causes a 4.5-fold increase in cytoplasmic volume fraction of peroxisomes. There is a concomitant induction of peroxisomal bifunctional enzyme and cytochrome P4504A, the enzyme markers associated with peroxisome proliferation. The spheroids also maintain expression of the peroxisome proliferator-activated receptor and preliminary data indicate that they are able to undergo replicative DNA synthesis in response to nafenopin. Hepatocyte spheroids will provide us with a model system for studying the early changes in rodent liver nongenotoxic carcinogenesis. PMID- 8405778 TI - Reliability of using fewer rabbits to evaluate dermal irritation potential of industrial chemicals. AB - To investigate the feasibility of using fewer than six animals for dermal irritation testing, a retrospective evaluation was conducted on data from 224 studies performed within the last 10 years using standard methodology. For each study, an irritation classification was made from primary dermal irritation indices calculated using irritancy data available for five, four, or three rabbits randomly selected from the original six-rabbit pool. The original six rabbit analyses resulted in classification of 49 non-irritants, 75 negligible irritants, 66 mild irritants, 24 moderate irritants, and 10 severe irritants. Agreement with the six-rabbit classification was obtained for five-, four-, and three-rabbit groups, respectively, as follows: for negligible irritants, 95, 85, and 69%; mild irritants, 85, 80, and 76%; moderate irritants, 88, 88, and 71%; severe irritants, 100, 90, and 90%. Differences in the dermal irritation indices between groups of six and five, four, and three rabbits were small, with an average difference no larger than 0.3. When disagreements in classification occurred, the test material was most often classified less severe. Using 90% or greater agreement as acceptable criterion for reducing the number of test animals, our results suggest that the use of less than six rabbits would not be suitable for evaluating dermal irritation based solely on a qualitative classification. However, the use of as few as three rabbits would be adequate to evaluate dermal irritancy based on quantitative primary dermal irritancy values. PMID- 8405779 TI - Tapetal effect of an azalide antibiotic following oral administration in beagle dogs. AB - An azalide antibiotic (CP-62,993) was administered at 100 mg/kg by oral gavage once daily for 35 consecutive days to 3 normal Beagle dogs (tapetal) and 3 Beagle dogs lacking a clinically apparent ocular tapetum (atapetal). The total dose delivered was approximately 100-fold the recommended clinical dose. Bilateral ophthalmoscopic changes were observed in the treated tapetal dogs on Day 36, consisting of mild to moderate tapetal decoloration with loss of the normal color change at the junction with the nontapetal fundus and muting of reflectivity of the normally highly reflective tapetum; treated atapetal and all control tapetal and atapetal dogs had no ophthalmoscopic changes. Microscopic examination of ocular tissue revealed rudimentary tapetal cell layers in the correct location in untreated, clinically atapetal eyes. Tapetal cells from treated tapetal and atapetal dogs were swollen and vacuolated, and contained intracytoplasmic, electron-dense debris but no recognizable tapetal rodlets. Lysosomal lamellar bodies were observed in the retinal ganglion cells of both treated groups and were neither enhanced nor reduced by the presence of a functional tapetum. Necrosis and inflammation were not observed in any ocular tissue. The altered ophthalmoscopic appearance of treated tapetal dogs was not influenced by the retinal changes because any effect on retinal transparency would have been seen in treated atapetal dogs. The decoloration and muting of reflectivity observed clinically in the tapetal fundus of dogs following prolonged exposure to high levels of CP-62,993 result from unique changes within the ocular tapetum itself and cannot be interpreted to be of consequence to nontapetal species including humans. PMID- 8405780 TI - Comparative carcinogenicity of 5,5-diphenylhydantoin with or without perinatal exposure in rats and mice. AB - Chronic toxicity and carcinogenicity studies of 5,5-diphenylhydantoin (DPH), were conducted in F344/N rats and B6C3F1 mice of each sex. The major objective of the study was to determine if incorporating exposure to DPH during the perinatal period, in addition to conventional exposure of animals for 2 years, enhances the sensitivity of the bioassay to identify the carcinogenic potential of chemical. The studies were designed to determine the toxic and carcinogenic effects of dietary DPH in rats and mice receiving; (1) the perinatal administration including exposure of maternal animals prior to breeding, through gestation, lactation, weaning, and continued dietary exposure of offspring to the age of 8 weeks followed by control diet for 2 years, (2) exposure for 2 years beginning at the age of 8 weeks, and (3) of combined perinatal/adult exposure to DPH (perinatal exposure to 8 weeks of age followed by the adult exposure for 2 years). During the perinatal period, rats were exposed to DPH at dose levels ranging from 63 to 630 ppm and adult exposure concentrations ranged from 240 to 2400 ppm in diet. In the mice, the perinatal exposure ranged from 21 to 210 ppm in both males and females. In the adult exposure portion of the mouse studies, the dietary levels ranged from 30 to 300 ppm in males and 60 to 600 ppm in females. A total of eight dose groups (including controls) were used with 60 animals in each group. The only effect of perinatal exposure alone on tumor rate was a marginal increase in the incidence of hepatocellular neoplasms in female mice. The adult exposure to DPH significantly increased the incidence of hepatocellular neoplasms in female mice. There were also marginal increases in the incidence of liver tumors in male rats exposed to high DPH dietary concentrations during the adult-only regimen. Combined perinatal and adult dietary exposure to 5,5-diphenylhydantoin confirmed the findings for the increased incidences of hepatocellular neoplasms in male rats and female mice, although combined exposure did not enhance these effects. However, in male mice, perinatal and adult exposure resulted in an increase in the incidence of hepatocellular neoplasms that was not seen when dietary exposure was limited to the adult period only. PMID- 8405781 TI - Lead and behavioral development in young herring gulls: effects of timing of exposure on individual recognition. AB - Lead exposure early in life affects behavioral, physiological, and intellectual development in humans and other animals. Recognition of parents or other caregivers and eventual bonding are essential aspects of behavioral development. In this paper young herring gulls, Larus argentatus, were used to examine the effect of timing of lead exposure on individual recognition behavior and development. Each of 60 1-day-old herring gull chicks was randomly assigned to a control group or to one of three treatment groups that received a single dose of lead nitrate solution (100 mg/kg) at Day 2 or 6 of age or the same total dose divided in thirds on Day 2, 4, and 6. Matched controls were injected with isotonic saline on the same schedules. Variations in individual recognition of human attendants were largely explained by age and status (lead versus control), and some variation was explained by day of injection (exposure regime). Using a feeding paradigm, the percentage responding to their caretaker compared to another person was higher (70% versus 38%), and occurred earlier in controls compared to lead-injected birds. Lead-injected birds required longer to respond initially, took longer to choose, moved less distance per time, and took longer to eventually reach the food. These results were all significant by GLM models and Kruskal-Wallis tests. Among lead-injected birds there was a disjunction of effect related to dosing schedule: birds injected on Day 6 chose more correctly but were slower to respond, indicating that these behavioral traits were differentially affected by timing of exposure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405782 TI - A modified spectrophotometric method appropriate for measuring cholinesterase activity in tissue from carbaryl-treated animals. AB - Inhibited cholinesterase in tissues of animals exposed to carbamate pesticides is known to reactivate readily, presenting considerable problems in the accurate assessment of cholinesterase activity in these tissues. Decarbamylation of cholinesterase is favored when the tissue samples are diluted and/or are incubated for an extended time. The present study was performed to identify modifications of the commonly used spectrophotometric assay for cholinesterase activity that would minimize spontaneous reactivation of enzyme activity. Those modifications included preincubation of concentrated tissue with concentrated chromogen (i.e., DTNB), dilution to final reaction volume immediately before measurement, and measurement of cholinesterase over a short period of time (5-10 min). The Ellman assay with and without modifications was performed using a microtiter plate reader on tissues from carbaryl-treated rats:undiluted plasma, diluted erythrocytes (1:25), minimally diluted erythrocytes (1:2), diluted brain (1:100), or minimally diluted brain (1:2). The results were compared to cholinesterase activities obtained using a radiometric method which employs minimally diluted tissue and short incubation times. The degree of cholinesterase inhibition for undiluted or minimally diluted tissue assayed by the modified method agreed with those obtained using the radiometric method. Even if the tissues were diluted immediately before assay, however, significant reactivation occurred by the time the first measurements were made by the conventional method. Furthermore, significant spontaneous reactivation may still occur using the modified method if the assay is run for more than 10 min. Use of this modified Ellman method will enable more accurate estimation of in vivo cholinesterase activity in animals treated with carbamates. PMID- 8405783 TI - Effects of 2-ethylhexanoic acid on reproduction and postnatal development in Wistar rats. AB - Reproductive toxicity of 2-ethylhexanoic acid (2-EHA) was studied in Wistar rats. The animals (24 animals per sex per group) were given 2-EHA as a sodium salt in drinking water at daily doses of 100, 300, or 600 mg/kg. Control animals received plain water. Male rats were exposed to 2-EHA for 10 weeks and females for 2 weeks prior to mating, both sexes during the mating period and females during the entire gestation and lactation period. 2-EHA caused a slight but dose-dependent decrease in fertility; time to mating increased at 300 and 600 mg/kg and even total infertility ensued. 2-EHA slightly decreased sperm quality in males. The spermatozoa were significantly less motile at 100 and 600 mg/kg and abnormal sperm occurred more frequently at the two highest dose levels. The average litter size was reduced by 16% in the dose group receiving 600 mg/kg. The birth weights of the pups were unaffected but the body weight gain was transiently slower during lactation at 600 mg/kg. Several pups appeared abnormal (kinky tail, lethargic, slightly paralyzed legs) and the physical development assessed by several landmarks (opening of eyes, eruption of teeth, hair growth) and reflexes (grip reflex, cliff avoidance) was delayed at 300 and 600 mg/kg. In another experiment, a single dose of 600 mg/kg 2-EHA was given to pregnant females by gavage on Gestational Day 4, 5, 6, or 7 and the number of implantations were counted on Gestational Day 10. Administration on Day 6 decreased the number of implantations and caused resorptions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405784 TI - Percutaneous penetration of N-nitrosodiethanolamine through human skin (in vitro): comparison of finite and infinite dose applications from cosmetic vehicles. AB - N-Nitroso compounds (nitrosamines) have been detected at the parts per billion level in a wide variety of matrices including industrial chemicals, pharmaceuticals, and food. Although N-nitrosodiethanolamine (NDELA) may be detected as an impurity in some cosmetic products, studies on NDELA absorption through human skin have been limited. A study to determine the extent of NDELA absorption following topical application was therefore undertaken to assist in the proper assessment of risk following unintended exposure. NDELA absorption was measured in vitro through human cadaver skin using isopropyl myristate (IPM) and generic prototype personal-care formulations (sunscreen and shampoo) spiked with [14C]NDELA. When applied as a finite dose at a concentration of 0.06% or lower, NDELA absorption was found to be a linear function of concentration. Total absorption at 48 hr ranged from approximately 35 to 65% of the dose and was formulation dependent (IPM > shampoo > or = sunscreen). Absorption occurred relatively rapidly from all formulations and peak rates of absorption were seen within the first 5 hr from the IPM and shampoo formulations. When applied as an infinite dose, total NDELA absorption followed a different rank order (shampoo > or = IPM > sunscreen) and evidence of barrier damage was seen with the shampoo formulation. PMID- 8405785 TI - Plasma concentrations in methyl isobutyl ketone-potentiated experimental cholestasis after inhalation or oral administration. AB - In studies of methyl isobutyl ketone (MiBK)-potentiated cholestasis induced by taurolithocholic acid (TLC) or manganese-bilirubin (Mn-BR) combinations, MiBK is usually given by gavage whereas industrial exposure to MiBK normally occurs by inhalation. The present study was conducted to verify if the route of administration could influence the potentiation. Male Sprague-Dawley rats were treated with MiBK for 3 days orally or by inhalation (4 hr/day). The minimal effective doses (MED) for potentiating both models of intrahepatic induced cholestasis were estimated to be 3 mmol/kg or 400 ppm for the oral or inhalation route, respectively. Groups of rats were treated with 0.5, 1, or 2 times the MED. Half of each group was sacrificed after the last MiBK administration to determine plasma concentrations of MiBK and its metabolites by gas-liquid chromatography. The other half was challenged 18 hr later with TLC (30 mumol/kg) or a combination of manganese (4.5 mg/kg) and bilirubin (15 mg/kg). Bile flow was measured from 15 to 135 min after the cholestatic challenge. Rats exposed to MiBK orally or by inhalation exhibited an enhanced diminution in bile flow that was dose-dependent. With dosages of 3 mmol/kg po or 400 ppm by inhalation or more, diminution in bile flow was significantly different from control values. Comparisons between maximal bile flow decrease and MiBK plasma concentration showed that the severity of the hepatotoxic response was dependent on the plasma MiBK concentration, irrespective of the route of administration. PMID- 8405786 TI - Coronary artery toxicity in beagle dogs given indolidan, a type III phosphodiesterase inhibitor. AB - The hemodynamic changes and cardiovascular lesions produced by a single iv dose of 2.0 mg/kg indolidan were evaluated in four beagle (two male, two female) dogs. Four additional dogs (two male, two female) served as vehicle controls. This was followed by a multidose study in which the same dose of indolidan or vehicle was given iv to dogs (4/group) on 4 consecutive days. Both studies were followed for 4 days postdose before termination. Clinical signs, mean arterial blood pressure, and heart rate were evaluated. Seven sections of the heart, approximately 25 sections of the coronary arteries, and 3 sections of the carotid, subclavian, spermatic or ovarian, renal, and femoral arteries were examined. Mean blood pressure was decreased 20 to 25% over a 24-hr period and heart rates were increased 40 to 50% after treatment and remained elevated for at least a 24-hr period in both single and multidose treated groups. The earliest lesions occurred in three of four treated dogs after a single intravenous dose of 2.0 mg/kg. The main lesion was sporadic in distribution and seen in the outer one-half of the smooth muscle media of the coronary arteries. Smooth muscle degeneration and necrosis were present, with little secondary inflammation. No lesions were observed in the peripheral arteries or the myocardium. The coronary arterial lesions observed after the multidose study were more extensive and severe. The lesions were seen in large extramural and large intramural coronary arteries. These were characterized by marked smooth muscle necrosis involving most of the media, destruction of the internal elastic membrane, and marked adventitial fibroplasia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405787 TI - Toxicokinetics of nickel in mice studied with the gamma-emitting isotope 57Ni. AB - The gamma-emitting isotope 57Ni was generated in a cyclotron to allow whole-body counting of laboratory animals dosed with nickel. 57NiCl2 was administered either orally by gastric intubation or by intraperitoneal injection to groups of mice in doses equivalent to the average human daily dietary nickel intake per mass unit. When given orally, the whole-body retention (WBR) was 0.02-0.36% of the administered dose at 45-75 hr. When given intraperitoneally, the WBR was 1-6% at 20-50 hr. After adjustment for the rapid excretion of systemic nickel, the intestinal absorption could be estimated to be 1.7-10%. The relative WBR did not vary with the magnitude of the dose within 0.05-5 mumol Ni/kg given orally or 0.005-0.5 mumol/kg given intraperitoneally. At 8 hr, the tissue concentration was highest in the kidneys, followed by the carcass, lungs, testicles, liver, and the spleen. After 20 hr, the highest concentrations were still found in the kidneys followed by the lungs, the liver, and the carcass. At 20 hr after oral administration, 50-70% of 57Ni retained in the body was within the carcass. The second highest nickel content was found in the kidneys, followed by the liver and lungs. Whereas nickel in the kidneys was rapidly excreted, the elimination from the lungs and liver was relatively slow, thereby, after 40 hr, resulting in a higher nickel content in the liver than that in the kidneys.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405788 TI - Leucovorin protection against repeated daily dose toxicity of trimetrexate in rats. AB - Repeated high doses of trimetrexate (TMX), a potent non-classical antifolate, have been administered as an experimental treatment for life-threatening Pneumocystis carinii infections in man. This therapy includes the coadministration of leucovorin, a reduced folate cofactor, to prevent antifolate toxicity in the host. The purpose of this investigation was to assess possible toxicologic sequelae of this combination regimen in an animal model. TMX at daily oral doses of 25, 35, and 45 mg/kg produced dose-related myelosuppression, thymic lymphoid depletion, seminiferous tubular atrophy, and degenerative lesions of the gastrointestinal tract. Mortality observed with TMX alone occurred earlier at higher doses and was specifically associated with severe degenerative enteropathy of the cecum. Oral leucovorin doses of 1, 5, 20, or 50 mg/kg administered twice daily, at the time of TMX administration and 6 hr later, protected against TMX lethality and target organ toxicity in a dose-related manner. Leucovorin was only partially protective against TMX-induced macrocytic anemia and the degree of protection was not dose-related. Leucovorin protection against cecal enteropathy was associated with increased DNA synthetic rates and higher mitotic activity of cecal epithelium than those in rats administered TMX alone. Importantly, the combination of daily administration of high dose TMX for 4 weeks with protective coadministration of leucovorin did not result in target organ toxicities that differed from TMX alone. PMID- 8405789 TI - Investigation of the effects of benomyl on rat nasal mucosa. AB - Benomyl [methyl 1-(butylcarbamoyl)-2-benzimidazolecarbamate, CAS Registry No. 17804-35-2] is a widely used agricultural fungicide. Previously, olfactory epithelial lesions were produced following a 45-day inhalation exposure to 50 and 200 mg/m3 benomyl. The present study, part of a range-finding study for a two generation reproduction study, was conducted to determine if the previously reported effects on the nasal mucosa are the result of systemic toxicity or attributable to the inhalation route of exposure. Groups of 10 7-week-old male Crl:CD BR rats were fed diets containing 0, 5000, 10000, or 15000 ppm benomyl for 32 days. Individual body weights and food consumption were determined weekly and on the last day of the study. After 32 days on test, rats were euthanatized by pentobarbital anesthesia and exsanguination and were examined for gross alterations. The nasal cavity was processed for pathological examination. Mean body weight gain was statistically significantly decreased during the first week of treatment and the overall test period (Days 0-32) at the two highest dose levels. A significant decrease in food consumption also was seen during test interval Days 0-7 for the two highest dose groups. In addition, statistically significant decreases in food consumption were observed at the Day 7-14 interval for the 15,000 ppm dose group and at the 21-28 and 28-32 intervals for the two highest dose groups compared with controls. No histopathological lesions were noted in the nasal epithelium of any of the control or benomyl-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405790 TI - A clarification on the Hot Springs Health Program. PMID- 8405791 TI - Cautionary notes on scientific misconduct. PMID- 8405792 TI - Pap smear process and quality assessment. PMID- 8405794 TI - Understanding authorship. PMID- 8405793 TI - Empathy in medical education. PMID- 8405795 TI - The Robert Wood Johnson Generalist Physician Faculty Scholar initiative. PMID- 8405796 TI - Does the author qualify for authorship? PMID- 8405797 TI - Entry of US medical school graduates into family practice residencies: 1992-1993 and three-year summary. AB - This is the 12th report prepared by the American Academy of Family Physicians (AAFP) on the percentage of each medical school's graduates entering family practice residency programs. Approximately 10.8% of the 15,466 graduates of LCME accredited US medical schools between July 1991 and June 1992 were first-year residents in family practice in October 1992. This compares to 10.3% the previous year. This is the first increase since 1989. Medical school graduates from publicly funded medical schools were more than twice as likely to be first-year residents in family practice in October 1992 than were residents from privately funded schools, 13.8% compared to 6.4%. The Mountain region reported the highest percentage of medical school graduates who were first-year residents in family practice programs in October 1992 at 18.3%; the Middle Atlantic and New England regions continued with the lowest percentages, 5.6% and 7.8%, respectively. Approximately one in two medical school graduates entering a family practice residency program as a first-year resident in October 1992 entered a program in the same state as that of his or her medical school of graduation. The percentages for each medical school have varied substantially from year to year since the AAFP has reported this information. The average percentage for each medical school for the last three years is reported. In addition, the number and percentage of graduates from colleges of osteopathic medicine who entered ACGME accredited family practice residency programs are reported. PMID- 8405798 TI - Results of the 1993 National Resident Matching Program. AB - The 1993 NRMP results reveal a 19% increase in positions filled in family practice residencies compared to 1992 (2,002 versus 1,658) and a 17% increase in positions filled by US seniors (1,636 versus 1,398). Similarly, 11% more positions were filled on July 1, 1993, than 1992 (2,798 versus 2,530). This confirms a trend begun in 1992 of increased interest in careers in family practice, reversing four years of declining interest from 1988 to 1991. Two hundred one fewer US seniors matched in internal medicine, while 36 more chose pediatrics. Of students entering residency training in the generalist disciplines, it is expected that 27% of the class of 1993 (LCME-accredited medical schools) will practice as generalists. An increase of 9% of first-year positions offered, and the development of 15% new ACGME-accredited family practice residency programs, could accommodate 20% of the nation's graduates of LCME- and AOA-accredited medical schools. With increasing interest in careers in family practice, increased support for the nation's family practice residency programs is critical. PMID- 8405799 TI - A review of constructivism: understanding and using a relatively new theory. AB - The purpose of this review paper is to familiarize family medicine educators with a relatively new educational theory, "constructivism." This theory is derived from the philosophical proposition that reality is constructed by the individual. According to the more traditional theory of "objectivism," knowledge exists in the world external to personal experience. Constructivist theory postulates that personal experience cannot be separated from knowledge. In analyzing the literature, the author found that constructivism can be viewed at the cognitive (individual) and social (community) levels. Cognitive constructivism maintains that individuals develop their own models of reality using personal experience and research-based data. Two key elements of cognitive constructivism with implications for family medicine educators are promoting student independence and active learning. Social constructivism maintains that individuals use their membership in a community to continually refine and shape their models of reality. By communicating with each other (for physicians, in the "conversation of medicine"), we test our constructs. Two key elements of social constructivism with implications for application by family medicine educators are promoting collaboration and peer teaching. PMID- 8405800 TI - Keeping the family in family practice: another way to attract medical students. PMID- 8405801 TI - Breast-feeding counseling practices of family practice residents. AB - BACKGROUND: Despite breast-feeding being the best newborn feeding method, the percentage of women in the United States who breast-feed has declined. Family physicians are in a unique position to counsel women about breast-feeding because their emphasis on continuity of care provides both prenatal and postnatal contact. Whether family physicians are trained sufficiently to offer such counseling is unknown. METHODS: A pretested questionnaire was distributed to all residents in 11 of 14 residency programs in North Carolina and Georgia. Data were analyzed with descriptive chi-square statistics and backward logistic regression to assess variables independently predictive of residents' counseling behaviors. RESULTS: One hundred fifty-five residents filled out usable surveys (response rate: 69%). The residents' primary teachers of breast-feeding were family physicians (34%) and nurses (33%). Most residents (93%) felt it was their personal responsibility to counsel women about breast-feeding; 67% believed their training was inadequate, and 48% stated they lacked necessary skills. Forty-one percent of residents counseled such women less than 50% of the time. Personal or spousal breast-feeding experiences gave residents more confidence in breast feeding anticipatory guidance (P < .001), comfort in teaching techniques (P < .001), and effectiveness in counseling (P < .009) but did not lead to improved counseling rates. Multivariate analysis showed that female residents, those with high confidence in their breast-feeding counseling skills, and residents that had adequate training in breast-feeding counseling instructed women more often about breast-feeding. CONCLUSIONS: Breast-feeding counseling skills are an important but neglected aspect of family practice clinical training. Personal breast feeding experience is an important indicator of breast-feeding counseling confidence and effectiveness, although it doesn't ensure that proper counseling skills are obtained. Family medicine training programs should incorporate breast feeding educational programs into their residency curricula. PMID- 8405802 TI - The effect of patient health status on physician practice style. AB - BACKGROUND: The recent development of health status measures now facilitates the study of how patient health influences physician practice style. METHODS: The health status of 150 new patients at a university primary care center was assessed using the Medical Outcomes Study Short-Form General Health Survey. Videotapes of the physician-patient encounter were analyzed with the Davis Observation Code to explore the effect of patient health status on physician practice style. RESULTS: Regression analyses demonstrated that better health was predictive of a greater portion of the visit being spent on physical examination and chatting and a smaller portion of the visit on history taking. Counseling was predicted by diminished patient mental health scores. Preliminary evidence was found for different practice styles based on patient characteristics such as sex, age, education, and income. CONCLUSIONS: Results suggest that the physician patient encounter is strongly influenced by health status. It will be crucial for future studies of physician-patient interaction to include an assessment of the patient's health status. PMID- 8405803 TI - Patient acceptance of two health status measures: the Medical Outcomes Study Short-form General Health Survey and the Duke Health Profile. AB - BACKGROUND: The Medical Outcomes Study Short-Form General Health Survey (MOS-SF) and the Duke Health Profile (DUKE) are brief, reliable, valid, and practical health status measures with potential applications in clinical research. We compared patient acceptance, ease of completion, and adequacy of capture (of the patient's self-perception of health) of these instruments in a primary care setting. METHODS: The MOS-SF, DUKE, and assessment questionnaires were administered to 79 patients in a university-based family practice. Patient acceptance of each instrument was assessed with Likert scale questions. Patients then compared the two instruments for relative ease of completion, preference, and completeness of capture. RESULTS: Both forms assessed each item well, but the DUKE scored statistically significantly better than the MOS-SF on four of six patient acceptance questions and both ease of completion items. There were no significant differences for capture items. CONCLUSIONS: Although both instruments are well accepted, investigators may favor the DUKE over the MOS-SF for situations in which patient acceptance or ease of completion is a key issue. PMID- 8405804 TI - Jurassic Park and the "gay gene": the new genetics seen through the distorting lens of the media. PMID- 8405805 TI - The revolution in communication for biologists--the internet. PMID- 8405806 TI - Extracellular matrix 4: the elastic fiber. AB - The elastic properties of many tissues such as the lung, dermis, and large blood vessels are due to the presence of elastic fibers in the extracellular space. These fibers have been shown by biochemical and ultrastructural analysis to be composed of two distinct components, a more abundant amorphous component and a 10 12 nm microfibrillar component, which is located primarily around the periphery of the amorphous component. The protein elastin makes up the highly insoluble amorphous component and is responsible for the elastic properties. Elastin is found throughout the vertebrate kingdom and possesses an unusual chemical composition rich in glycine, proline, and hydrophobic amino acids, consonant with its characteristic physical properties. The 72-kDa biosynthetic precursor, tropoelastin, is secreted into the extracellular space where it becomes highly cross-linked into a rubber-like network through the activity of the copper requiring enzyme lysyl oxidase. Analysis of the elastin gene has demonstrated that hydrophobic and cross-linking domains are encoded in separate exons and that there is significant alternative splicing, resulting in multiple isoforms of tropoelastin. The elastin gene promoter contains many potential binding sites for various modulating factors indicative of a complex pattern of transcriptional regulation. The microfibrils contain several proteins, including fibrillin, and probably act as an organizing scaffold in the formation of the elastin network. There appears to be a fibrillin gene family in which each protein contains multiple repeats of a motif previously found in epidermal growth factor and a second motif observed in transforming growth factor beta 1-binding protein. Mutations in the fibrillin gene located on human chromosome 15 have been strongly implicated as the cause of the Marfan syndrome. PMID- 8405807 TI - Biogenesis: number mysticism in protein thinking. AB - Historically, great minds have been tantalized by the idea that integers contain hidden, subtle meanings that could give us deep insights into natural (and supernatural) phenomena. Numerological analysis has been used in religion, mythology, and the sciences. In the field of proteins, integers played a stimulating role during early struggles to unravel structure, but they ultimately proved constrictive and misleading. In contrast, the introduction of imaginary (or complex) numbers into the algebra and numerical analysis of ligand-protein affinities can open new perspectives into such interactions. PMID- 8405808 TI - Astrocytes in infectious and immune-mediated diseases of the central nervous system. AB - The central nervous system (CNS) can be invaded and damaged by a variety of microbes. The host response to such injury involves CNS cells, and in many cases hematogenous cells also. Recent experiments indicate that astrocytes, macroglial resident cells of the CNS, play key roles in this process. The astroglial production of trophic factors and elimination of neurotoxins are likely to fulfill important protective and reparative functions during CNS infection. In addition, astrocytes could, in concert with microglial cells, regulate the recruitment and activity of infiltrating hematogenous cells through their expression of cytokines, proteases, protease inhibitors, adhesion molecules, and extracellular matrix components. Although previous experiments suggested that astrocytes might initiate inflammatory demyelinating disease by presenting CNS antigens to autoreactive immune cells, current evidence points against such a detrimental activity. In view of the generally beneficial role of astrocytes, impairments of astroglial function by microbes or host-derived factors have the potential to contribute to neurologic disease. Diseases in which this pathogenetic process may be relevant include HIV-1-associated cognitive/motor complex and spongiform encephalopathies. PMID- 8405809 TI - Ischemia: from acidosis to oxidation. AB - Organ damage can occur quickly when blood flow is compromised. Lactic acidosis has long been associated with such ischemia, and many physicians assume that organ damage is caused by this acidosis. However, reviewing the literature related to hypoxia and ischemia reveals little data to support the concept of acidosis as damaging to tissue. In contrast, recent studies indicate that the acidosis is actually protective, even during reperfusion when cellular damage may occur. Reperfusion is accompanied by generation of free radicals and other reactive species that can damage proteins, membranes, and nucleic acids, supporting an emerging view that implicates these reactive species in the actual tissue damage. The critical targets of the damaging species are not known, but reaction with key enzymes and structural proteins could certainly disrupt organ function. Cellular proteins are oxidatively modified during reperfusion, in part by metal-catalyzed oxidation in which cellular iron plays a key role. Metal catalyzed oxidation of proteins may be important in the pathogenesis of other disorders, including the potentially blinding disease, retinopathy of the premature. PMID- 8405810 TI - Aspartylglycosaminuria: protein chemistry and molecular biology of the most common lysosomal storage disorder of glycoprotein degradation. AB - Aspartylglycosaminuria (AGU) (McKusick 20840) is the most common disorder of glycoprotein degradation caused by the failure of lysosomes to digest the protein to-carbohydrate linkage of Asn-linked glycoproteins. During the past few years there has been significant progress in our understanding of both the protein chemistry and molecular biology of glycosylasparaginase (EC 3.5.1.26) as well as the molecular changes underlying the storage disease AGU that results from deficiency of this lysosomal hydrolase. Modern clinical assays have been developed for the diagnosis and carrier detection of this disease. Detailed structure, substrate specificity, mechanism of action, and a part of the active site of glycosylasparaginase have been defined. Molecular biology of glycosylasparaginase has progressed rapidly and already some mutations in the glycosylasparaginase gene resulting in AGU have been identified. Evolutionary aspects based on sequence data indicate a mechanistic relationship between mammalian glycosylasparaginases and bacterial/plant asparaginases. PMID- 8405811 TI - Trans-sialidase: a unique enzyme activity discovered in the protozoan Trypanosoma cruzi. AB - Trypanosoma cruzi, the agent of Chagas' disease, an ailment characterized by a progressive chronic fibrotic myocarditis and degeneration of tissues that are innervated by the autonomic nervous system, is a voracious sialic acid eater from glycoconjugates of the surrounding medium. This is accomplished through an active trans-sialidase residing on the surface membrane of the trypomastigote stage, which is the parasite form that invades vertebrate cells. The existence of the enzyme was proposed and established only 7 years ago and yet a flood of information on the subject is already available. Trans-sialidase is able to reversibly transfer sialic acid alpha(2-->3)-linked to an external Gal beta from the host cell surface sialoglycoconjugates to a terminal Gal beta of an appropriate acceptor on the parasite surface. In the absence of an acceptor, the enzyme acts as a hydrolase transferring sialic acid to water. Trans-sialidase belongs to a highly heterogeneous gene family of surface molecules sharing with each other and with bacterial neuraminidases variable degrees of nucleotide sequence homology and common motifs. It has been proposed that sialylation of the parasite surface catalyzed by trans-sialidase is necessary for successful invasion of the host cell, but the evidence available is still indirect. Another function could be a protection from lysis by the alternative pathway of complement while the parasite is circulating in the acute phase of the disease. PMID- 8405812 TI - Human immune response to polydimethylsiloxane (silicone): screening studies in a breast implant population. AB - Although initially it was thought that polydimethylsiloxane (silicone) was biologically inert, recent published studies have demonstrated varying levels of IgG antibody reactive with this structure in humans. The objective of our study was to determine whether silicone implanted in humans results in a measurable immune response directed against a 3700 mol wt hydroxyl terminated silicone molecule and whether that response could be correlated with the level of presumed silicone exposure as inferred by clinical history. In a blind study, sera from 111 patients, with and without breast implants, were sent to a laboratory using an enzyme-linked immunosorbent assay to determine specific anti-silicone IgG antibody levels. Test results showed that patients with implants demonstrated statistically significant elevation in anti-silicone antibodies compared with the unimplanted control groups. The highest anti-silicone antibody levels were measured in implanted women with either frank implant ruptures or leakage of their silicone gel implants. PMID- 8405813 TI - Vitamin A status and immunoglobulin G subclasses in rats immunized with tetanus toxoid. AB - Vitamin A deficiency severely compromises the magnitude of the primary and secondary antibody response to tetanus toxoid (TT) but does not impair the development of immunologic memory. To further characterize this immunodeficiency in antibody production, we have quantified the immunoglobulin G (IgG) subclasses (IgG1, IgG2a, IgG2b, and IgG2c) during the primary and secondary response to TT in normal, vitamin A-deficient, and retinol-repleted rats. In the primary response in normal rats, anti-TT IgG1 and IgG2b predominated. In vitamin A deficient rats the production of anti-TT IgG2b was severely impaired, with little change in either IgG1 or IgG2a. In the secondary response vitamin A-deficient rats produced low levels of all anti-TT IgG subclasses. However, when vitamin A deficient rats were repleted with retinol 2 days before reimmunization their secondary anti-TT IgG response was normal both in magnitude and IgG subclass distribution. This result implies that although vitamin A deficiency during the primary antibody response impaired anti-TT IgG2b production, it did not inhibit Ig heavy chain recombination or the differentiation of lymphocytes that formed memory B cells for each subclass; furthermore, these cells were activated in the secondary response after vitamin A status was improved. Thus, these experiments further support the concept that memory cell formation remains normal during vitamin A deficiency despite low levels of antibody production. PMID- 8405814 TI - Use of receptor antibodies to demonstrate membrane glucocorticoid receptor in cells from human leukemic patients. AB - Anti-peptide antibody to the human glucocorticoid receptor (GR) was produced and used to demonstrate that a subset of the GR population resides in the plasma membrane of human leukemic cells. Characterization of the antibody with intracellular GR (iGR) showed its ability to shift [3H]triamcinolone acetonide labeled GR (4S protein) from two human leukemic cell lines to a higher density in sucrose gradients; Western and autoradiographic analysis of affinity-labeled ([3H]dexamethasone 21-mesylate) receptor revealed an immunoreactive and competitively labeled band of 94 kDa. CCRF-CEM cell membrane GR (mGR) resolved as a > 7S protein on density gradients and immunoselected cell surface protein labeled by whole cell biotinylation or affinity-labeling with [3H]dexamethasone 21-mesylate was approximately 145 kDa, demonstrating that mGR was larger in size than iGR, as has been shown previously for the mGR of mouse lymphoma cells. Analysis of mGR in lymphocytes of leukemic patients and the CCRF-CEM cell line indicated differences in levels of expression as shown by FACS and immunocytochemical analyses. We are currently using this system to study the correlation between the quantity of membrane-resident GRs and the glucocorticoid induced lytic response, a relationship previously shown in the murine (S-49 cell) system. PMID- 8405815 TI - Nitric oxide synthesis inhibition induces leukocyte adhesion via superoxide and mast cells. AB - Recent work has demonstrated that inhibition of nitric oxide production with various nitric oxide synthesis inhibitors (L-NAME, L-NMMA) initiate leukocyte adhesion to postcapillary venules. The objective of this study was to elucidate the mechanism (or mechanisms) that promote the L-NAME-induced leukocyte response. Intravital microscopy was used to examine 25-40 microns venules in the rat mesentery. Nitric oxide synthesis was inhibited with L-NAME and leukocyte adhesion was observed over the first 60 min. The fourfold increase in leukocyte adhesion was independent of alterations in venular red blood cell velocity. The adhesion was superoxide-mediated inasmuch as superoxide dismutase (SOD) abolished the rise in leukocyte adhesion associated with nitric oxide synthesis inhibition. Ketotifen, a mast cell stabilizer, also abolished the rise in leukocyte adhesion induced by L-NAME. Histology revealed that mast cell degranulation occurred only in animals treated with L-NAME but not in animals pretreated with SOD or ketotifen. This observation suggests that mast cells become activated in the absence of nitric oxide production and superoxide contributes to the mast cell activation. The L-NAME-induced leukocyte adhesion could be reproduced by infusing hypoxanthine/xanthine oxidase (a superoxide generating system) or compound 48/80 (an activator of mast cells) and both responses were attenuated by ketotifen. These data suggest that inhibition of nitric oxide synthesis results in a superoxide and mast cell-dependent leukocyte adhesion. PMID- 8405816 TI - Biochemical teamwork: revealing the niacin-tryptophan relationship. PMID- 8405817 TI - Out-of-hospital symptomatic supraventricular arrhythmias. Epidemiological aspects derived from 10 years experience of the Florence Mobile Coronary Care Unit. AB - The epidemiologic features and the relative incidence of symptomatic supraventricular tachycardias in out-of-hospital settings are unknown. Rhythm disturbances account for 20% of the interventions performed by the Florence Mobile Coronary Care Unit (MCCU). Between November 1979 and December 1989, the MCCU rescued 1239 patients with recent onset (less than 24 hours) symptomatic supraventricular arrhythmias. 809 had atrial fibrillation, 376 paroxysmal supraventricular tachycardia (PSVT), 36 atrial flutter and 18 different atrial dysrhythmias. Women showed an overall predominance, more evident in patients with PSVT, and the incidence of the arrhythmias increased with age. Preexisting heart disease was more frequent in atrial fibrillation (41.1%) and atrial flutter (33.4%) in comparison to PSVT (27.6%). Similarly, a higher incidence of associated cardiovascular events (AMI, acute coronary insufficiency, pulmonary edema) was found in patients with atrial fibrillation and atrial flutter. Palpitations were the main complaint in each group, however, in atrial fibrillation and atrial flutter they were frequently associated with chest pain or dyspnea. PMID- 8405818 TI - [Nodal reentry tachycardia: short- and long-term effectiveness and safety of a selective ablation technique of the slow pathway]. AB - BACKGROUND: Various ablation methods have been proposed in the last few years in order to find a radical solution for atrioventricular nodal reentrant tachycardia. The first techniques were surgical, followed by modulation of the fast pathway, causing a prolongation of the P-R interval, the latter involving a 2 to 10% atrioventricular block risk. To reduce this risk, slow pathway ablation was then suggested, with the objective of abolishing atrioventricular reentry. The aim of our study was to evaluate how frequently the recording of peculiar slow potentials was possible in patients with atrioventricular nodal reentrant tachycardia, and to assess short-and long-term efficacy of an ablation technique involving the use of these potentials as electrophysiologic markers. METHODS: One hundred and eighty-eight patients with typical atrioventricular nodal reentrant tachycardia were studied (mean age 47 +/- 18 years). Radiofrequency ablation was guided by peculiar slow potential recordings; when this was not possible, fast pathway ablation, or slow pathway ablation guided only by anatomic markers, were performed. RESULTS: Potentials with peculiar electrophysiologic characteristics were found during sinus rhythm in the median posterior region of the septum, anteriorly to the coronary-sinus ostium, in 92% of patients. These characteristics included: low amplitude; the fact that they occupy the first part of the interval between the atrial and ventricular electrogram; their amplitude diminishes and disappears with increased frequency of atrial stimulation and/or with atrial extrastimulus. Typical atrioventricular nodal reentry tachycardia was no longer inducible in any patient at the end of the procedure with a median of 2 radiofrequencies application per patient. No II or III degree atrioventricular block was caused when ablation was guided by slow potential recordings. During an attempt at fast pathway ablation a complete atrioventricular block was caused in 1 patient. One hundred and eighty-four patients remained asymptomatic during a follow-up of 2 to 24 months; no one showed either a modification of atrioventricular conduction if compared to that found at hospital discharge or proarrhythmic effects. Four patients had one atrioventricular nodal reentrant tachycardia recurrence and a second successful ablation was performed in 2 of these 4 patients. CONCLUSIONS: Peculiar slow potentials, that can be used as electrophysiologic markers for slow pathway ablation, were recorded in the medio posterior region of the septum in the majority of patients. The fact that this technique, using slow potential as an electrophysiologic marker, was successful in all patients, with very few recurrences and with no serious complications (no II or III degree atrioventricular block) makes it trustworthy and safe. PMID- 8405819 TI - [Cardiac and vascular hypertrophy in juvenile borderline hypertension: echocardiographic and ultrasonographic study]. AB - BACKGROUND: High resolution ultrasonography is a noninvasive technique that allows us to investigate the cardiovascular system, in particular the wall thickness and the lumen diameter of the arteries, with accuracy and reproducibility. METHODS: We measured the intima-media thickness of the common carotid artery (CCA) and of its bifurcation (BIF) in 20 borderline hypertensive (age 24 +/- 4 years) and in 20 normotensive subjects (age 24 +/- 4 years), as a control group. Both carotid axes were scanned from different views (anterior, lateral, posterior) on a transversal and longitudinal section using a high resolution steerable linear array of 5 MHz. Carotid diameter and thickness were measured in the longitudinal section. CCA parameters were assessed 20 mm caudally to the flow divider. RESULTS: In borderline patients blood pressure (147.8 +/- 10.5/90.7 +/- 6.6 mmHg) and left ventricular mass index (102.5 +/- 15.3 g/m2) were significantly higher than in normotensive subjects (blood pressure 120.5 +/- 11.5/78.0 +/- 5.4 mm Hg; left ventricular mass 90.5 +/- 14.3 g/m2). The intima media thickness of both the CCA and BIF was significantly higher in borderlines than in normotensives (CCA 0.6 +/- 0.08 vs 0.4 +/- 0.05 mm, p < 0.001; BIF 0.7 +/ 0.08 vs 0.5 +/- 0.08, p < 0.001). In the whole population there was a statistically significant correlation between the carotid wall thickness and the left ventricular mass. CONCLUSIONS: Our data show that ultrasonography provides direct evidence that in young borderline hypertensives the increased left ventricular mass is associated with vascular hypertrophy. PMID- 8405820 TI - [Silent myocardial infarction caused by acute carbon monoxide poisoning]. AB - This report describes a 63 year-old man who suffered an acute myocardial infarction after carbon monoxide exposure. When evaluated in the Emergency Room the patient was completely conscious and did not experience any chest pain. The electrocardiogram showed non-specific T wave abnormalities in inferolateral leads. The only sign consistent with a possible myocardial involvement was a slight increase in serum CPK. The electrocardiogram taken 12 hours later revealed a Q-wave inferior myocardial infarction, and CPK levels showed a typical elevation in the following 24 hours. The coronary angiography, performed about two weeks after admission, documented multiple obstructions in the main coronary branches. In the presence of a reduced coronary reserve, the onset of a prolonged myocardial ischemia might have been secondary to a decreased oxygen transport capacity of the blood, which lead to a decreased amount of oxygen available to the tissues. According to their experience, the authors emphasize the importance of a careful electrocardiographic and enzymatic monitoring of all patients in the first hours after CO exposure, because the typical chest pain may be absent. PMID- 8405821 TI - [Acute myocardial infarction in a young adult with multiple coronary aneurysms resulting form Kawasaki disease]. AB - A young male patient with no risk factors for atherosclerotic disease suffered from an acute myocardial infarction at the age of 22 years, and was subsequently found to have multiple coronary artery aneurysms by coronary angiography. The transthoracic echocardiography was unable to identify coronary anomalies, whereas the transesophageal approach did show aneurysmatic lesions of the left anterior descending artery. These could have been caused by a previous episode of Kawasaki disease, a pathological finding that should be considered in any young adult presenting with proximal discrete coronary artery aneurysms. PMID- 8405822 TI - [Endothelins: a new family of peptides with biologic effects. Which is their physiopathologic role?]. PMID- 8405823 TI - [Coronary angioplasty in multivessel disease]. PMID- 8405824 TI - ["Physiologic" aging of the cardiovascular system]. PMID- 8405825 TI - [Reevaluation of physiopathologic and clinical aspects of syncope in sick sinus syndrome]. PMID- 8405826 TI - [Provocative tests of myocardial ischemia: the current scenario]. PMID- 8405827 TI - [Nuclear methods in cardiology]. PMID- 8405828 TI - [An unusual coronary variant]. PMID- 8405829 TI - [The patient's consent]. PMID- 8405830 TI - [The end of cardiomyopathies]. PMID- 8405831 TI - The impact of meteorological factors on the onset of myocardial infarction in the coastal region of middle Dalmatia. AB - The influence of mediterranean climate on the onset of acute myocardial infarction (AMI) has been investigated in a study group comprised of 1306 AMI patients from the coastal part of middle Dalmatia (Croatia) admitted to the two Split hospitals in the 1981-1987 period. AMI incidence during the southern wind (0.62 per day) was significantly greater than during the northern wind (0.50 per day; p < 0.05), as well as during all non-southern-wind days (0.49 per day; p < 0.01). The coefficients of partial correlation showed a significant association of AMI incidence with increased air temperature (p < 0.05) and relative humidity (p < 0.05). Cross-correlation showed a positive link of AMI incidence to increased air temperature four days before, and on the day of the incident (p < 0.05). Analysis of the synoptic situations revealed characteristics of unstable weather in the days with 4 or more AMI patients. The results suggest that during the southern wind or the passing atmospheric front, coronary patients should stay at home, avoid both physical and psychological stress, and take antianginal drugs more frequently. PMID- 8405832 TI - [Late ventricular potentials in the post-infarct patient. A follow-up at 4 years]. AB - We performed a prospective study to evaluate the prognostic significance and the natural history of late ventricular potentials (LPs) in 209 patients (165 males and 44 females; mean age 59.8 +/- 10 years) who survived acute myocardial infarction. Signal-averaged electrocardiograms (SA-ECGs) were performed before hospital discharge (16 +/- 5 days) and after four years (mean follow-up 42 +/- 7 months). SA-ECGs were processed using a 40 Hz high-pass bidirectional filter. Duration of "filtered" QRS (normal value < 120 msec), duration of the low amplitude signals (n.v. < 39 msec) and last 40 msec voltage of the QRS complex (n.v. > 20 microV) were measured. LPs were defined as the presence of two or more abnormal values. In addition, 24-hour Holter monitoring was performed in all patients, and left ventricular ejection fraction (LVEF) was determined by scintigraphy in 120 (57.4%). Sixty patients (28.7%) had LPs before hospital discharge (group 1), and 149 (71.3%) had normal SA-ECGs (group 2). During the follow-up period there were 10 arrhythmic events, 7 of which were sudden deaths, and three cases of sustained ventricular tachycardia. SA-ECG was repeated in 141 patients (68%). The mean values of SA-ECG's parameters did not change significantly between the two controls, and the correlation was good for all of them. Despite this, spontaneous normalization of SA-ECGs occurred in 21 patients (60%) and the subsequent appearance of LPs was seen in 13 (12%); in these latter, the SA-ECG's parameters measured before hospital discharge were "borderline" and significantly different from those who did not change. The sensitivity of SA-ECG as a predictor of arrhythmic events was 80% and the specificity 74%. Patients with arrhythmic events had a longer filtered QRS (126 +/- 33 vs 103 +/- 12 msec; p < 0.001), longer duration of the low-amplitude signals (57 +/- 23 vs 32 +/- 11 msec; p < 0.001), lower voltages (17 +/- 8 vs 36 +/- 24 microV; p < 0.001), and, moreover, higher peak CK values, lower LVEF and higher value of Lown modified class. In conclusion, SA-ECG confirms its value in identifying patients at risk of arrhythmic events after myocardial infarction. SA-ECG recordings taken before the discharge can be used to predict serial changes during follow-up. PMID- 8405833 TI - [The effect of physical exercise on the response to exertion in the elderly]. AB - BACKGROUND: A decrease in adaptation to exertion has been observed as age progresses. Although this decline may also be affected by factors such as health conditions and age, physical inactivity related to sedentary behaviour plays a dominant role. METHODS: In order to evaluate the influence of physical activity on cardiovascular response to exertion in the elderly, 4 groups of 22 subjects each were submitted to maximal electrocardiographic exercise test on a cycloergometer (multistage program with 30 Watts x 3 min. steps). All subjects were male. The composition of the groups was as follows: 1) veteran long distance runners (mean age: 71 +/- 5.4); 2) sedentary veterans (mean age: 69.8 +/- 3.9); 3) young long distance runners (mean age: 25.4 +/- 4.3); 4) sedentary young adults (mean age: 25.8 +/- 3.9). The endurance athletes, well fitted to competition, had been practicing sport activity for at least 3 years. RESULTS: Heart rate, arterial systolic and diastolic blood pressure were recorded; mean blood pressure and double product were calculated at baseline and at the climax of the stress test; furthermore, total and maximal watts were recorded. For each of the parameters, Student's t test for non-paired observations were used to evaluate statistical differences amongst the four groups. The most interesting result arises in the comparison between veteran long distance runners and sedentary young adults: between the two groups no statistically significant differences in workload, expressed as total watts (1649.55 +/- 296.32 vs 1650.00 +/- 446.32; p = NS) and maximal watts (175.91 +/- 19.19 vs 173.18 +/- 24.38; p = N.S.), were observed. On the contrary, highly significant differences in both total (p < 0.01) and maximal (p < 0.01) watts were noticed by comparing long distance runners and sedentary subjects of the same age. CONCLUSIONS: These data support the hypothesis that the progressive reduction in physical activity, which is usually observed in aging, is the major determinant of exercise deconditioning in the elderly. PMID- 8405834 TI - [Acute rejection: the diagnostic and therapeutic problems]. AB - Acute allograft rejection is still a leading cause of both early and late mortality and morbidity after cardiac transplantation. Besides cell-mediated acute rejection, a "humoral" form exists which is more frequent in younger patients in the early postoperative period, and less likely to respond to the usual therapy. Cyclosporine therapy has deeply affected the clinical, laboratory and histological aspects of acute rejection. Though endomyocardial biopsy remains the most reliable diagnostic tool, other noninvasive procedures are of great value for a final diagnosis; among these are electrocardiography and two dimensional echocardiography, which are the two elective techniques for post transplantation follow-up in children. The laboratory and immunological assays, on the contrary, share a very low specificity. In conclusion, the diagnosis of acute rejection is still a clinical one, though supported by laboratory and histological evidence. In our experience, the natural evolution of many mild and mild-moderate rejection episodes toward regression does not support an excessive prophylaxis or an early treatment of symptom-free acute rejection. In addition, treatment of rejection must be personal and specific to each patient. PMID- 8405835 TI - ["Incomplete closure" of the mitral valve: the relationships to valvular regurgitation and to the morphofunctional characteristics of the left ventricle]. AB - OBJECTIVES: The present study was designed in order to evaluate the prevalence of mitral regurgitation in patients with the "incomplete mitral leaflet closure" echocardiographic pattern, to verify whether the amount of "incomplete mitral leaflet closure" is related to the severity of mitral regurgitation and, last, to verify the relation between the "incomplete mitral leaflet closure" and left ventricular morphology and function. METHODS: We studied 80 patients (14 patients with dilatative cardiomyopathy, 26 patients with coronary artery disease, and 40 patients with hypertensive heart disease or aortic valve disease) showing the "incomplete mitral leaflet closure" pattern, retrospectively selected from a population composed of 1700 consecutive patients routinely examined in our echocardiographic laboratory. In all patients we evaluated the presence and the severity of mitral regurgitation, the morphological and functional parameters of the left ventricle, the systolic diameter of the mitral annulus, the distance between the point of mitral leaflet coaptation and the annular plane, and the incomplete mitral closure area, assuming the last two parameters as indexes of the severity of incomplete closure of the mitral valve. RESULTS: We observed the presence of mitral regurgitation in 51 out of 80 patients (64%). The valvular insufficiency was considered mild in 78% of the patients. We observed no significant difference between patients with mitral regurgitation and without, as regards the diameter of the mitral annulus, the distance between the point of mitral leaflet coaptation and the annular plane, and the incomplete mitral closure area in different types of heart diseases. The incomplete mitral closure area and the diameter of the mitral annulus showed a significant, although not elevated, correlation with the severity of the mitral regurgitation (r = 0.36 and r = 0.32, respectively). The severity of mitral regurgitation showed significant correlations with all of the left ventricular morphological and functional parameters evaluated. Finally, we observed significant correlations between the incomplete mitral closure area and all of the morphological and functional parameters of the left ventricle. CONCLUSIONS: On the basis of the results obtained we can conclude that: 1) the "incomplete mitral leaflet closure" pattern does not appear to be a highly specific marker of mitral regurgitation, 2) this pattern appears to be related to the morphology and function of the left ventricle, and 3) the severity of the incomplete mitral valve closure is more easily evaluated by a parameter that takes into account the numerous factors acting on the mitral apparatus, that is the incomplete mitral closure area. PMID- 8405836 TI - [The many faces of parasystolic rhythm]. AB - BACKGROUND: Parasystole is the expression of a pacemaker that is protected from, and thus independent of, the dominant rhythm. The arrhythmia is not always associated with the 3 classic signs: variable coupling, mathematically related interectopic intervals, and fusion beats. A large amount of experimental and clinical data have pointed out several atypical phenomena that make the recognition of parasystole difficult. This especially occurs in the presence of influence exerted from sinus impulses upon the parasystolic rhythm. METHODS: A pattern of ventricular parasystole was evident throughout a 24-hour Holter recording obtained from a 55-year-old female. The following data were analyzed: a) distribution of ectopic complexes; b) parasystolic cycle duration; c) regularity of parasystole, as assessed by means of the variation index. In some sections of the tracing in which an electrotonic influence (modulation) exerted by the sinus rhythm on the parasystolic rhythm appeared as very likely, phase response curves were constructed in order to express the time-dependent modulation effected by sinus impulses. RESULTS: Parasystole occurred in two separate periods, lasting 4 hours and 90 minutes, respectively. On both occasions, the arrhythmia was apparently precipitated by a brief episode of atrial tachycardia. Phases of regular parasystole, as well as periods of irregular modulated parasystole, were observed. The distribution of ectopic complexes was at times typical for concealed bigeminy (intervening beats always in odd numbers), but on other occasions reflected the even variant of concealed bigeminy where the intervening beats conform to the formula 1 + n (n being zero or an odd number). Some couplets of identical ectopic complexes were also observed. CONCLUSIONS: The study shows that several different expressions of parasystolic rhythm may be present within the same tracing. A minimal or absent modulation results in the classical picture of parasystole; when a mild modulating influence is present, the typical pattern of modulated parasystole ensues, whereas a strong modulation leads to disappearance of the typical features of parasystole and manifestation of concealed bigeminy. Finally, supernormal modulation is responsible for the occurrence of couplets. PMID- 8405837 TI - [Myocardial infarct and sudden death with angiographically undamaged coronary arteries]. AB - Clinicopathologic findings in two young adults, who complained of ischemic cardiac arrest by ventricular fibrillation and of myocardial infarction complicated by cardiogenic shock respectively, are described. At coronary angiography, coronary arteries appeared normal. In both cases, detailed pathologic examination of the coronary arteries disclosed a focal eccentric atherosclerotic plaque in the proximal descending coronary artery, where selective coronary angiography had failed to reveal filling defects. These observations suggest a more critical attitude in evaluating angiographically normal coronary arteries in patients with myocardial infarction or aborted sudden death, and underline the possibility of arterial lumen stenosis underestimation, especially in the presence of eccentric plaque, with likely compensatory ectasia of the plaque-free wall segment. PMID- 8405838 TI - [Prolonged angina after the administration of a synthetic PGE2 derivative]. AB - We describe the case of a 39-year-old woman, heavy smoker, who received 500 micrograms i.m. of Sulprostone, a synthetic PGE2-derivative, to induce pregnancy termination. Sulprostone is usually administered either to cause abortion in preparation of an instrumental operation or to induce delivery after the intrauterine death of the fetus. This drug has a dilating effect on the cervix uteri and stimulates the uterus muscles. After about fifteen minutes the patient experienced a constrictive chest pain which progressively worsened and spread to the upper limbs. The pain disappeared for a short period and then recurred with greater intensity, accompanied by bradycardia and hypotension. The ECG showed sinus bradycardia, second- and third-degree atrioventricular block, S-T segment elevation in the inferior leads and reciprocal depression in the anterior leads. Intravenous nitroglycerin therapy induced a rapid reduction of the clinical symptoms and changes in the ECG. There was no increase in cardiac enzymes. The exercise test, the cold pressor test and the ECO-dipyridamole test were negative. The patient refused to undergo the ergonovine test and coronary angiography. We hypothesize that the Sulprostone either had a dipyridamole-like effect or that it induced a paradoxal coronary spasm. PMID- 8405839 TI - [The classification of cardiomyopathies: is a revision opportune?]. PMID- 8405840 TI - The role of Doppler echocardiography in the diagnosis of constrictive pericarditis. PMID- 8405841 TI - [The "anthology syndrome" and ethics viewed by heretics]. PMID- 8405842 TI - [The vital myocardium and the echo-nitrate test]. PMID- 8405843 TI - [Reviewers: arbiters or consultants?]. PMID- 8405844 TI - Myenteric plexus destruction alters morphology of rat intestine. AB - BACKGROUND: It has been shown previously that myenteric plexus destruction by benzalkonium chloride (BAC) increased villus height, crypt depth, and muscle thickness, suggesting that these neurons influence intestinal morphology. A nonspecific trophic effect of BAC, intraluminal stasis, and inflammation resulting from the chemical treatment could also be causes for these changes. Our goals were to (1) show that the morphological sequelae of BAC treatment are caused by myenteric plexus removal and not the factors listed above, and (2) determine whether segmental myenteric plexus removal alters morphology elsewhere in the small intestine. METHODS: Six groups of rats were studied: control, chemical denervation (3 mmol/L BAC), surgical denervation, intraluminal stasis produced by partial obstruction, chemical inflammation (5% acetic acid), and surgical inflammation (serosa removal only). Tissue for histological study was taken from the treated segment, 15-20 cm proximal to the treated segment, and 5 10 cm distal to the treated segment 28 days after treatment. RESULTS: Chemical and surgical denervation reduced the number of myenteric neurons by 94% and 98%, respectively. Denervation had a direct effect on morphology; it increased villus height, crypt depth, and muscle thickness in the treated and proximal segments, but only muscle thickness was increased in the distal segment. The other treatments had minimal morphological sequelae. CONCLUSIONS: Segmental myenteric plexus removal alters the mucosa in the treated and proximal segments but influences muscle thickness throughout the intestine. PMID- 8405845 TI - Modified Doppler flowmetry in the splanchnic circulation. AB - BACKGROUND: Absolute measurements of blood flow by Doppler ultrasonography are subject to substantial error. A simplified method of Doppler flowmetry using conventional duplex Doppler equipment was studied to establish its validity in determining short-term changes in flow in the splanchnic circulation in portal hypertension. METHODS: Two simple indices were measured: peak velocity averaged over two cardiac cycles and vessel diameter. The relationship between peak and mean velocity was studied using a Doppler phantom. The feasibility of studying relevant vessels and the constancy of regional flow was determined by performing Doppler flowmetry in a basal state twice within 2 hours. RESULTS: Constancy of relationship between peak and mean velocities was confirmed (r > 0.98). Examination was successful in the majority of 28 consecutive patients with portal hypertension for superior mesenteric, main portal, right portal, and paraumbilical (when patent) veins, but was successful in only 54% for the hepatic artery. Constancy of basal flow was found in 11 selected patients when group data were considered, but in individuals, substantial and unpredictable variance was found. CONCLUSIONS: Modified Doppler flowmetry appears valid in the assessment of short-term changes in splanchnic blood flow of patients with portal hypertension. Individual variance in baseline flow is identified as a potential problem and is further reason for caution in the interpretation of measurements of Doppler flow. PMID- 8405846 TI - Mechanisms of HCl-induced lowering of intracellular pH in rabbit esophageal epithelial cells. AB - BACKGROUND: For acid injury to occur in esophageal epithelium, extracellular HCl must lower intracellular pH. Therefore, we sought to define the mechanisms for translation of low extracellular pH (pHo) into low intracellular pH (pHi). METHODS: To define the mechanisms, primary cultures of rabbit esophageal epithelial cells were loaded with the pH-sensitive fluorescent dye, 2'7'' bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF), which enabled pHi to be recorded by microfluorimetry. RESULTS: Lowering pHo to 6.0 with HCl caused pHi to decline at 0.07-0.08 units/min and produced an average cellular H+ load of 3-6 mmol/L at 1 minute and of 11 mmol/L at 5 minutes. This degree of acidification was primarily attributable to increased H+ entry via a 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS)-sensitive Clo-dependent mechanism because DIDS or exposure to Clo-free solution blocked cell acidification. Furthermore a contribution to low pHi at low pHo of abolition of acid extrusion via Na+/H+ and Na(+)-dependent Cl-/HCO3- exchangers was sought but not established because exposure at low pHo to either 5-(N-ethyl-N isopropyl)amiloride (EIPA) or Na(+)-free solution + EIPA increased the degree of acid loading over untreated cells. CONCLUSIONS: Low pHo results in low pHi in esophageal cells primarily because of increased H+ entry via a DIDS-sensitive, Clo-dependent mechanism consistent with the known acid-loading Na(+)-independent Cl-/HCO3- exchanger. PMID- 8405847 TI - Effects of intrasphincteric botulinum toxin on the lower esophageal sphincter in piglets. AB - BACKGROUND: The toxin of Clostridium botulinum (BoTx) inhibits the release of acetylcholine from nerve terminals and causes paralysis of skeletal muscle. The present study examined the hypothesis that BoTx may have a similar effect on gastrointestinal smooth muscle. METHODS: Baseline lower esophageal sphincter (LES) pressures were obtained in five piglets, and normal saline was injected endoscopically into the LES. One week later, LES pressure was measured again, followed by injection of BoTx into the LES. After another week, LES pressure was measured again. RESULTS: Compared with a baseline LES pressure of 8.2 +/- 1.5 mm Hg, LES pressure decreased to 3.2 +/- 1.0 mm Hg after BoTx injection, a reduction of about 60% (P < 0.01). By contrast, LES pressure did not change significantly after normal saline injection. The animals showed no evidence of toxicity. Data from other experiments showed that after injection with toxin, the LES responds normally to bethanechol and pentagastrin but displays a paradoxical response to edrophonium and cholecystokinin. CONCLUSIONS: BoTx is a potent inhibitor of resting LES tone. Its relatively specific anticholinergic effect may help clarify the role of cholinergic and noncholinergic pathways in the regulation of gastrointestinal sphincters. PMID- 8405848 TI - GLUT2 is the transporter for fructose across the rat intestinal basolateral membrane. AB - BACKGROUND: The exact roles of disaccharidases and GLUT5 in the brush border membrane and GLUT2 in the basolateral membrane in the absorption of fructose across the intestine have not been fully determined. This paper describes characterization of fructose transport across the jejunal basolateral membrane using isolated membrane vesicles. METHODS: Transport of fructose was measured using rapid filtration of vesicles. Luminal perfusion in vivo with glucose and fructose before vesicle preparation was used to assess modulation of GLUT2 activity. Western blotting measured the abundance of GLUT2 in the membrane. RESULTS: The maximal rate of transport for fructose was 1100 pmol/mg protein/s and the Michaelis constant was 16 mmol/L. Fructose and glucose could completely inhibit the transport of each other. Perfusion of the intestinal lumen with fructose or glucose saline for 4 hours produced a fourfold increase in maximal fructose transport. CONCLUSIONS: These data indicate that the one transport protein, GLUT2, is responsible for moving both fructose and glucose out of the enterocyte across the basolateral membrane under basal conditions. The activity of this, or a closely related carrier, is rapidly upregulated by the presence of hexoses in the intestinal lumen, explaining the potentiation of fructose absorption by luminal glucose and obviating any need to involve apical disaccharidases. PMID- 8405849 TI - Oral 5-aminosalicylic acid for inflammatory bowel disease in pregnancy: safety and clinical course. AB - BACKGROUND: Oral 5-aminosalicylic acid (5-ASA) has proven an effective maintenance therapy of ulcerative colitis and may also be useful in Crohn's disease, but its safety in pregnancy has not been established. The present study therefore examined the course and outcome of pregnancies in patients with inflammatory bowel disease who continued to take oral 5-ASA. METHODS: Ten patients with ulcerative colitis and 7 patients with Crohn's disease with a total of 19 pregnancies were studied while they were receiving 5-ASA. All patients were previously in remission on 5-ASA, at a mean dose of 1.7 g/day (range, 0.8-2.4 g/day). They continued taking the drug without a change in dose and were followed up throughout their pregnancies and postpartum. RESULTS: Eighteen pregnancies resulted in full-term delivery. No fetal abnormalities were found at delivery, and there were no clinical or biochemical abnormalities in the neonatal period. Four patients had a relapse. One patient required a colectomy but carried on to a full-term pregnancy. One patient had a miscarriage, but she had miscarried on four previous occasions before taking 5-ASA. She subsequently had a successful pregnancy on 5-ASA. CONCLUSIONS: Oral 5-ASA appears to be safe for the management of inflammatory bowel disease during pregnancy. PMID- 8405850 TI - Evaluation of liquid yeast-derived sucrase enzyme replacement in patients with sucrase-isomaltase deficiency. AB - BACKGROUND: No enzyme replacement therapy exists for patients with congenital sucrase-isomaltase deficiency (CSID). A by-product of the manufacture of baker's yeast is a liquid preparation containing high sucrase activity. The aim of the present study was to investigate the activity and stability of this preparation and its effect on breath hydrogen excretion and gastrointestinal symptoms after sucrose ingestion in 14 patients with CSID. METHODS: The homogeneity of yeast sucrase was studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and its activity was measured. Stability at various temperatures and pH ranges and in the presence of gastric aspirate, pepsin, and bovine serum albumin was assessed. Fourteen patients with CSID underwent double-blind placebo-controlled breath tests with yeast sucrase. They then completed an 8-week dose response study that used different enzyme concentrations while consuming a sucrose containing diet. RESULTS: Liquid yeast sucrase is highly glycosylated, contains no lactase activity, and is stable at 4 degrees C and over a wide range of pH. Pepsin digestion of the enzyme in vitro can be blunted by bovine serum albumin and by increasing the pH. Yeast sucrase reduces breath hydrogen excretion in patients with CSID who are given a sucrose load (P < 0.001) and allows most patients to consume a sucrose-containing diet. CONCLUSIONS: Liquid yeast sucrase offers effective enzyme replacement therapy for patients with CSID. PMID- 8405851 TI - Intracerebroventricular neuropeptide Y increases gastric and pancreatic secretion in the dog. AB - BACKGROUND: Neuropeptide Y (NPY), a centrally located neurotransmitter, is known to increase appetite in fasted and satiated animals. In addition to evaluating NPY's effect on eating behavior, this study was intended to determine whether intracerebroventricular (ICV) NPY would have an effect on canine gastric and pancreatic secretion. METHODS: Four dogs were prepared with cerebroventricular guides and gastric and pancreatic fistulas. ICV and intravenous NPY was administered during intragastric titration of a glucose and peptone meal. During this study, gastric and pancreatic secretion was measured, as well as insulin levels and pancreatic polypeptide (PP). An additional set of four dogs were prepared with esophageal fistulas and cerebroventricular guides, and the effect of ICV NPY on sham feeding was studied. RESULTS: ICV NPY significantly increased sham feeding, meal-stimulated gastric and pancreatic secretion, basal gastric acid, pancreatic bicarbonate, insulin levels, and PP. Vagotomy blocked the effect of ICV NPY on gastric acid secretion in a urethane-anesthetized rat model with acute gastric fistula. CONCLUSIONS: ICV NPY increased sham feeding, gastric and pancreatic secretion, insulin levels, and PP in the dogs. NPY's effect on gastric secretion was blocked by vagotomy in a rat model. NPY should be considered a candidate mediator of cephalic phase secretion. PMID- 8405852 TI - Variability in the risk of major gastrointestinal complications from nonaspirin nonsteroidal anti-inflammatory drugs. AB - BACKGROUND: We have assessed the extent to which the risk of serious gastrointestinal complications from nonaspirin nonsteroidal anti-inflammatory drugs (NANSAIDs) varies with the age and sex of recipients, use of aspirin or alcohol, administration by the oral or rectal route, and dose and choice of drug. METHODS: A case-control study was performed with prospective recruitment of cases of gastrointestinal bleeding or ulcer perforation and age- and sex-matched controls. Information on preadmission drug use obtained by structured interview. RESULTS: Six hundred forty-four patients and 1268 controls were recruited. The odds ratio for upper gastrointestinal complications in users compared with nonusers of NANSAIDs increased with age: < or = 59 years, odds ratio 2.0; 60-79 years, odds ratio 3.0; > or = 80 years, odds ratio 4.2; and was higher in women (5.4) than in men (1.9). There was a linear dose-response curve that was steeper in women than in men. Combined exposure suggested additive risks: NANSAIDs and aspirin, odds ratio 6.7; NANSAIDs and alcohol, odds ratio 6.0 NANSAIDs by the oral route were associated with an odds ratio of 2.3, compared with 11.4 with rectal administration. Piroxicam was associated with the highest risk, odds ratio 4.8; and ibuprofen the lowest risk, odds ratio 0.7. CONCLUSIONS: A number of factors can alter the risk of major gastrointestinal complications with NANSAIDs and need to be considered when individual prescribing decisions are made. PMID- 8405853 TI - A novel mechanism for disposing of effete epithelial cells in the small intestine of guinea pigs. AB - BACKGROUND: We previously showed that at the villus tips in the small intestine of guinea pigs effete enterocytes are not simply exfoliated into the lumen but phagocytosed by subepithelial macrophages, leaving only a thin apical cell portion in the epithelial lining. The aim of the present study is to investigate the fate of these apical pieces of enterocytes. METHODS: The ileum of guinea pigs was perfusion-fixed and processed for transmission and scanning electron microscopic observation. RESULTS: The apical cytoplasmic plates were found to be pushed by neighboring enterocytes and protruded from the epithelial surface, finally being pinched off into the lumen. In this process observed at the villus tips, the junctional complexes between the apical cytoplasmic plate and the adjacent enterocytes were preserved until the pinching-off of the plate. Luminal cell elements revealed a rich existence of cup-shaped or spherical cell fragments covered with microvilli; nuclei were never observed in the luminal fragments. CONCLUSIONS: The findings in the small intestine of the guinea pig are the first to account for the mechanism of the epithelial barriers being preserved while apoptotic enterocytes drop out at the tips of the villi. PMID- 8405854 TI - Helicobacter pylori infection: independent risk indicator of gastric adenocarcinoma. AB - BACKGROUND: Helicobacter pylori has been implicated as a possible etiologic factor in gastric cancer. This case control study was performed to determine the association between H. pylori and gastric cancer, taking into account the possibility of confounding by other background factors. METHODS: Sera were collected from 112 incident case patients with gastric cancer and 103 control patients with nongastroenterological diseases, who were frequency-matched with respect to age and sex. Immunoglobulin G antibodies to H. pylori were identified using the HM-CAP immunoassay (Enteric Products Inc., Wesbury, NY). RESULTS: The prevalence of H. pylori seropositivity was significantly higher (P = 0.002) among case patients than control patients. The odds ratio (OR) was 2.60 (95% confidence interval, 1.35-5.02). The increased OR associated with H. pylori infection was confined to tumors with a noncardia location (OR, 3.06) and men (OR, 4.27). OR increased with decreasing age at cancer diagnosis to reach 9.33 in patients < 60 years of age. Multivariate logistic regression analysis was used as control for potential confounding, but the elevated OR associated with H. pylori infection remained significantly increased. CONCLUSIONS: The results support the hypothesis of H. pylori infection as an independent risk indicator of gastric cancer. PMID- 8405855 TI - Increased expression of muscular neural cell adhesion molecule in congenital aganglionosis. AB - BACKGROUND: Neural cell adhesion molecule (NCAM) is down regulated during morphogenesis and innervation of cardiac and skeletal muscle. In mature muscle, its reexpression over the entire sarcolemma occurs in response to denervation or paralysis of muscle and in some myopathies. No information is available regarding NCAM expression in human enteric muscle either in health or in disease. Our aim was to test whether NCAM is present in nerves and muscle of normal infant bowel and to determine how its expression is altered in congenital aganglionosis. METHODS: Using immunocytochemistry for light microscopy, we compared the pattern of distribution of NCAM in congenitally aganglionic colon with that in colon from age-matched controls. RESULTS: In normal colon, NCAM immunoreactivity was seen in ganglion cells and nerve fibers throughout the gut wall and, more weakly, on the inner border of the circular muscle. In aganglionic bowel, there was a marked increase in NCAM expression in muscle, particularly that of the muscularis mucosac and characteristic hypertrophied nerve bundles of the intermuscular zone and submucosa displayed immunoreactivity for NCAM. CONCLUSIONS: Abnormal expression of NCAM is, thus, a feature of congenital aganglionosis and is likely to be associated with neuromuscular dysfunction within the affected colon. PMID- 8405856 TI - Spasmolytic polypeptide is a major antral peptide: distribution of the trefoil peptides human spasmolytic polypeptide and pS2 in the stomach. AB - BACKGROUND: The regional differences in the distribution of the trefoil peptides pS2 and human spasmolytic polypeptide (hSP) within the stomach are not known. The aim of this study was to gain insight into these functionally obscure molecules by characterizing their distribution. METHODS: Tissue from gastrectomy specimens removed for peptic ulceration was examined to chart the distribution of hSP and pS2, using immunohistochemistry and in situ hybridization to chart their messenger RNAs (mRNAs). RESULTS: Colocalization of pS2 and hSP was noted in the gastric foveolar and surface epithelium throughout the stomach. In the gastric antrum and pylorus, an extremely strong hSP mRNA signal was present within pyloric-type glands; strong hSP immunostaining was also seen at this site and in mucous-neck cells. Neither pS2 mRNA nor pS2 peptide were shown within the deep portions of the pyloric glands. Within areas of intestinal metaplasia, a few goblet cells immunostained for pS2 and putative ulcer associated cell lineage was seen with a pattern of trefoil peptide localization similar to the ileum. CONCLUSION: The detailed function of these trefoil peptides is unknown, but their distribution suggests involvement in repair-enhancing mechanisms. hSP may be an important antral peptide and both of these peptides may play a specific reparative role. PMID- 8405857 TI - Effect of D-glucose on intestinal permeability and its passive absorption in human small intestine in vivo. AB - BACKGROUND: Based on studies in animals, it has been proposed that carrier mediated D-glucose absorption markedly enhances passive permeability of the jejunal mucosa, allowing the majority of D-glucose absorption to proceed passively. In this study, we evaluated this hypothesis in the human jejunum in vivo. METHODS: Using the constant perfusion, nonabsorbable marker technique, permeability of jejunal mucosa was assessed by measuring the ratio of diffusion rates of urea/L-xylose and mannitol/L-xylose. Passive D-glucose absorption was quantitated using L-glucose and mannitol as probes for D-glucose. RESULTS: Addition of D-glucose to perfused solutions did not change the diffusion ratios, indicating that D-glucose has no effect on the size of channels for passive diffusion across the jejunal mucosa. The fraction of total D-glucose absorption that could be attributed to a passive mechanism averaged 5%. In the human ileum in vivo, we detected no evidence of passive D-glucose absorption. CONCLUSIONS: Carrier-mediated D-glucose absorption does not increase passive permeability of human jejunal mucosa to solutes with molecular radii between 2.6 and 4.0 A. The amount of D-glucose absorbed passively from the human jejunum is trivial compared with the overwhelmingly dominant mechanism, carrier-mediated transport. Our results do not support the concept that sodium-dependent nutrient transport increases tight junction permeability. PMID- 8405858 TI - DNA mapping of colorectal neoplasms: a flow cytometric study of DNA abnormalities and proliferation. AB - BACKGROUND: There is some evidence that neoplastic development and progression evolve through a multistep process associated with hyperproliferation and genetic alterations. Therefore, changes of proliferation and of cellular DNA content within the adenoma-carcinoma sequence were studied. METHODS: Using a "mapping" procedure, 12 adenomas and 18 carcinomas were analyzed flow cytometrically and histologically. In addition, normal mucosa adjacent to and distant from the tumors was assessed in the same way. RESULTS: Of 59 adenomatous fractions, 35.6% (n = 21) were aneuploid, whereas the incidence of aneuploidy was 63.5% (54/85) in the carcinomatous sites. Additional tetraploidies were identified in 5 (8.5%) and 13 (15.3%) adenomatous and carcinomatous samples, respectively. Cell proliferation, as determined by the percentage of S-phase cells, was significantly (P < 0.001) higher in the carcinomatous specimens (14.8% +/- 0.8%; mean +/- SEM) than in the adenomatous ones (8.1% +/- 0.7%). It decreased to normal mucosa adjacent to (5.1% +/- 0.5%) and distant (5.3% +/- 0.6%) from the neoplasms. DNA mapping of the tumors revealed both distinct regions and extended areas of aneuploidy and tetraploidy. There is evidence from the mapping data that aneuploid populations arise at a single focus of the adenoma and expand over large areas before a subpopulation of cells acquires the capacity of invasion. CONCLUSIONS: These data showing consecutive DNA content abnormalities within the colorectal adenoma-carcinoma sequence provide support for genomic instability and clonal evolution as important events of tumorigenesis and progression. PMID- 8405859 TI - Hepatic mitochondrial energy production in rats with chronic iron overload. AB - BACKGROUND: Iron overload results in impaired hepatic mitochondrial oxidative metabolism. The current experiments evaluated the effects of iron overload on enzyme activities in the mitochondrial electron transport chain, on hepatic adenine nucleotide levels, and on hepatocellular oxygen consumption. METHODS: Hepatic iron overload was produced in rats using dietary carbonyl iron. Hepatic adenine nucleotides were assessed after freeze-clamping, mitochondrial enzyme activities and oxygen consumption were measured in isolated mitochondria, and oxygen consumption in isolated hepatocytes was determined. RESULTS: At a mean hepatic iron concentration of 4630 micrograms/g, there were no changes in reduced nicotinamide adenine dinucleotide (NADH)-cytochrome c reductase activity (complex I-III), but there was a 35% reduction in succinate-cytochrome c reductase activity (complex II-III), and a 70% decrease in cytochrome c oxidase activity (complex IV). With mild iron loading (2060 micrograms/g), there was a 28% decrease in hepatic adenosine 5'-triphosphate (ATP) levels with no change in adenosine 5'-diphosphate (ADP) or adenosine 5'-monophosphate (AMP) levels, whereas, at a higher hepatic iron concentration (3170 micrograms/g), there was a 40% reduction in ATP levels, a 22% decrease in ADP levels, with no change in AMP levels. There was a 48% reduction in oxygen consumption in isolated iron-loaded hepatocytes. CONCLUSIONS: Chronic iron overload decreases hepatic mitochondrial cytochrome c oxidase activity, hepatocellular oxygen consumption, and hepatic ATP levels. PMID- 8405860 TI - Serology of acute exacerbation in chronic hepatitis B virus infection. AB - BACKGROUND: Liver injury in many patients with chronic hepatitis B is sporadic and is often characterized by acute exacerbations alternating with relatively normal liver function. The aim of this study was to perform detailed serological and biochemical analysis during periods of active liver disease to better understand the mechanisms responsible for the cyclic nature of liver injury. METHODS: A series of serum samples from 19 hepatitis B e antigen (HBeAg)-positive patients were analyzed for alterations in serum hepatitis B virus (HBV) DNA, HBeAg, anti-HBe production, and HBeAg-specific immune complex formation before, during, and after spontaneous acute exacerbations of liver injury. RESULTS: This analysis revealed significant correlations between increasing levels of serum HBV DNA, HBeAg, HBeAg-specific immune complexes, and liver injury. These results suggest that increases in viral replication and accumulation of viral proteins in the serum and intracellularly and the subsequent immune response play an important role in initiating acute exacerbations of liver injury in chronic hepatitis B infection. CONCLUSIONS: We propose that production of the secreted and cellular forms of the nucleoprotein reaches a threshold level and elicits specific immune responses, which mediate liver injury. PMID- 8405861 TI - Manometry of esophageal varices: comparison of an endoscopic balloon technique with needle puncture. AB - BACKGROUND: A noninvasive technique of pressure measurement in esophageal varices using an endoscopic balloon has been shown to be reliable in vitro. In the present study, this method was tested in vivo. METHODS: Thirty-seven pressure measurements in esophageal varices were performed in 34 patients by two independent operators (A and B) using an endoscopic balloon and compared with measurements performed by needle puncture by a third operator (C). RESULTS: Three measurements performed with the endoscopic balloon were rejected because they were noninterpretable. Measurements performed by A and B correlated well (correlation coefficient, 0.90); interobserver variability (r) was 0.88. Of 37 punctures performed for pressure measurements, 4 resulted in bleeding and 8 measurements were rejected as uninterpretable. Regression analysis showed a good correlation between the needle puncture and balloon techniques for pressure measurements performed by both operators (y = 5.3 + 1.0x, r = 0.8; y = 6.2 + 0.9x, r = 0.8), and analysis of variability showed a measurement bias of -5.3 +/- 4.1 and -4.1 +/- 3.5 cm H2O. No significant difference in variceal size measured with the endoscopic balloon or endoscopic forceps was found. CONCLUSIONS: The endoscopic balloon allows measurement of pressure in esophageal varices without hazard to the patient; in addition, it may be used to assess the varix size. PMID- 8405862 TI - Impaired reactivity of the peripheral vasculature to pressor agents in alcoholic cirrhosis. AB - BACKGROUND: Studies of the in vivo vascular reactivity of the peripheral circulation to pressor agents in cirrhosis have produced conflicting results, possibly because of changes in mean arterial pressure that make it difficult to clearly separate peripheral and central effects. The aim of the present study was to assess the reactivity of the forearm circulation to pressor agents in vivo without activating central control systems. METHODS: Forearm blood flow was measured by venous occlusion strain gauge plethysmography in the basal state and during the infusion of subpressor doses of norepinephrine and angiotensin II into the brachial artery in 10 male patients with well-compensated alcoholic cirrhosis and 10 male age-matched controls. Plasma renin activity and aldosterone and angiotensin II concentrations were assayed. Forearm and systemic sympathetic nervous system activity was estimated using a norepinephrine spillover technique. RESULTS: Basal forearm blood flow, renin angiotensin aldosterone system activity, and forearm sympathetic nervous system activity were similar in both the control and cirrhotic groups. The cirrhotic patients showed an impaired response to both norepinephrine and angiotensin II. CONCLUSIONS: There is impaired reactivity of the peripheral vasculature to pressor agents in cirrhosis, indicating that the control of vascular tone is disturbed even in well-compensated cirrhosis. PMID- 8405863 TI - Nucleotide sequence analysis of the precore region in patients with fulminant hepatitis B in the United States. AB - BACKGROUND: A precore defective hepatitis B virus (HBV) mutant unable to produce hepatitis B e antigen (HBeAg) has been associated with fulminant hepatitis B. We have studied the etiologic contribution of precore mutants among North American patients with this disorder. METHODS: We studied 39 patients with fulminant hepatitis B. The precore and proximal core regions of HBV from 37 of 39 patients were sequenced. RESULTS: Four patients (10.8%) harbored nonsense mutants likely to produce an HBeAg negative HBV infection; two such mutants had a G to A substitution at position 1896, one lost the precore initiation codon, and one harbored a stop codon immediately downstream of the precore initiation codon. Recovered sequences from seven additional patients displayed silent or missense mutations in these regions. All delta coinfected patients harbored known wild type strains of HBV. A significantly poorer survival was associated with antibody to HBe positivity and presence of nucleotide substitutions in the precore/core region. CONCLUSIONS: The prevalence of precore mutations in 37 patients from the United States was lower than reported elsewhere; only two patients were found to have the G to A transition mutation in the precore region at position 1896. We conclude that HBeAg negative HBV mutants do not play a predominant etiologic role among North American patients with fulminant hepatitis B. PMID- 8405864 TI - An evaluation of human recombinant alpha interferon in patients with metastatic gastrinoma. AB - BACKGROUND: Metastatic gastrinoma is becoming increasingly recognized in patients with Zollinger-Ellison Syndrome. The mean 5-year survival of these patients is < 20%. Chemotherapeutic regimens are of limited benefit. The aim of this study was to evaluate the use of interferon in these patients because a preliminary report suggested it might be effective. METHODS: The efficacy and toxicity of interferon was assessed in 13 consecutive Zollinger-Ellison syndrome patients with liver metastases. Patients were treated with human recombinant alpha interferon (5 million IU, subcutaneously [SC]) daily and followed up at 3-month intervals with multiple imaging studies. At each follow-up, toxicity of therapy was assessed and fasting serum gastrin concentrations were obtained. RESULTS: No patient showed a reduction in tumor size at any follow-up. One patient died after 2 months. At 6 months, six patients (46%) had stable tumor size in the liver, although new bone metastases developed in one patient. Three patients showed stable disease for up to 21 months. Changes in serum gastrin correlated with tumor response at 6 months. All patients developed some side effects of therapy. Thirty-one percent required dose reduction, and one patient (8%) had to have interferon therapy interrupted briefly. CONCLUSIONS: These results fail to define a therapeutic role for interferon in the treatment of metastatic gastrinoma. PMID- 8405865 TI - Cisapride improves gallbladder contractility and bile lipid composition in an animal model of gallstone disease. AB - BACKGROUND: The hepatic secretion of supersaturated bile and gallbladder stasis are key events in cholesterol gallstone formation. The therapeutic value of cisapride, a prokinetic agent, was assessed in ground squirrels on a 1% cholesterol diet. METHODS: Biliary lipid secretion was measured directly and bile salt pool size assessed by isotope dilution ([14C]cholic acid). Gallbladder contraction was measured in vitro in response to cholecystokinin (CCK). RESULTS: Cholesterol-fed animals had a combined hepatic secretory defect (a 53% decrease in bile salt secretion and also a 31% increase in cholesterol secretion). Adding cisapride restored bile salt secretion to control levels but did not affect cholesterol secretion. In cholesterol-fed animals, the cholesterol saturation index of gallbladder bile more than doubled and cholesterol crystals developed; cisapride markedly reduced cholesterol saturation, thus preventing crystal formation. Gallbladder contractility, measured in vitro in response to CCK, decreased 23% in animals on the 1% cholesterol diet; cisapride restored the CCK dose-response curve to normal. The bile salt pool as assessed by isotope dilution was similar in all groups. CONCLUSIONS: Thus, lithogenic bile develops in this model because of reduced bile salt secretion and increased cholesterol secretion. Cisapride renders biliary lipid composition towards normal by enhancing gallbladder (and possibly intestinal) motility and cycling of the bile salt pool, thereby increasing bile salt secretion. PMID- 8405866 TI - Different feedback regulation of hepatic cholesterol and bile acid synthesis by glycodeoxycholic acid in rabbits. AB - BACKGROUND: To explore the sexual difference in the feedback regulation of hepatic bile acid synthesis, glycodeoxycholic acid (GDCA) was administered to 15 male and 14 female rabbits. METHODS: After bile diversion, GDCA equivalent to the hepatic bile acid influx was infused intraduodenally. Biliary cholic acid output represented bile acid synthesis. Hepatic 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase and cholesterol 7 alpha-hydroxylase activities and steady state messenger RNA (mRNA) levels were determined. RESULTS: GDCA inhibited bile acid synthesis less in female than in male rabbits. Hepatic HMG-CoA reductase activity decreased 39% in males, but increased 48% in females. Hepatic cholesterol 7 alpha-hydroxylase activity decreased similarly in males and females, and mRNA levels decreased 86% in males but were unchanged in females. CONCLUSIONS: (1) Total bile diversion stimulated both hepatic cholesterol and bile acid synthesis by activating the rate-controlling enzymes and increasing mRNA levels. (2) GDCA decreased mRNA levels of HMG-CoA reductase and cholesterol 7 alpha-hydroxylase in males, but mRNA levels did not decrease in females. (3) Bile acid synthesis was sustained in females because continued biosynthesis of cholesterol provided a substrate for cholesterol 7 alpha-hydroxylase and stimulus for enzyme formation. PMID- 8405867 TI - Gallbladder mucin, arachidonic acid, and bile lipids in patients who develop gallstones during weight reduction. AB - BACKGROUND: Arachidonic acid (AA) and hydrophobic bile salts (BS) stimulate gallbladder mucin (GBM) secretion, which is thought to be an essential step in gallstone pathogenesis. The present study was performed to evaluate the relationship between AA, BS, and GBM in patients who develop gallstones following weight reduction. METHODS: Eleven patients who underwent gastric bypass, developed symptomatic gallstones, and then underwent cholecystectomy were evaluated. Gallbladder bile was obtained for analysis during each procedure. Matched patients who did not develop gallstones following gastric bypass served as controls. RESULTS: GBM increased in every patient who developed stones (mean increase: 5000%). The largest increase was observed soon after gastric bypass, and this declined curvilinearly with time. Gallbladder bile cholesterol was initially elevated but then rapidly declined before increasing back to pregastric bypass levels after weight loss was complete. No significant changes in phosphatidylcholine molecular species (including AA) or BS composition were observed following weight reduction. Concentrations of cholesterol, phospholipids, and changes in [AA] over time were each a linear function of [BS]. No relationship between GBM and any of these bile constituents was apparent. CONCLUSIONS: These observations strongly suggest that increases in GBM, which occur with gallstone formation in humans, are not the result of alterations in biliary AA or BS composition. PMID- 8405868 TI - Biliary papillomatosis with the point mutation of K-ras gene arising in congenital choledochal cyst. AB - Biliary papillomatosis is a rare entity. A case of biliary papillomatosis associated with congenital choledochal cyst and intrahepatic gallstones is reported here. Percutaneous transhepatic cholangioscopy revealed multiple papillary lesions of the right intrahepatic duct and the common bile duct. Microscopically, the papillary mucosal lesion showed papillary proliferations of bile duct epithelial cells with mild atypia. Furthermore, a point mutation at codon 12 of the K-ras oncogene was found in the papillary lesion. To the best of our knowledge, this is the first case of biliary papillomatosis arising in congenital choledochal cyst. Although the pathogenesis of biliary papillomatosis in our case was unclear, biliary irritation associated with choledochal cyst may be related to biliary papillomatosis with point mutation at codon 12 of K-ras gene. PMID- 8405869 TI - Endoscopic stent placement for internal and external pancreatic fistulas. AB - Stents have been effectively used for various pancreatic conditions. Pancreatic fistulas, however, have traditionally been considered a surgical disease, and if the fistula does not respond to conservative measures, an operation is usually performed. Stents were placed endoscopically in five consecutive patients who presented with pancreatic fistulas that did not respond to conservative management. Fistulas resolved in all patients after endoscopic stent placement, and after 14-30 months of follow-up, none has recurred. The cases comprise two patients with pancreaticocutaneous fistula and one each with pancreaticopleural, pancreaticoperitoneal, and pancreaticocholedochal fistula. The need for an operation can be obviated in many patients with internal and external pancreatic fistulas. PMID- 8405870 TI - Diarrhea in patients with the acquired immunodeficiency syndrome. PMID- 8405871 TI - Reduce, reuse, and recycle: shedding light on shedding cells. PMID- 8405872 TI - Showdown at the tight junction. PMID- 8405873 TI - Immunization with naked DNA: a sexy technique. PMID- 8405874 TI - Varicella virus in achalasia. PMID- 8405875 TI - Ornithine decarboxylase: an oncogene? PMID- 8405876 TI - Is the long-term outcome after posttransfusion hepatitis benign? PMID- 8405877 TI - Acinar cell loss following duct ligation in the rat pancreas: necrosis or atrophy? PMID- 8405878 TI - Hyperbaric oxygen and perineal Crohn's disease: a follow-up. PMID- 8405879 TI - Gastrin, the enterochromaffinlike cell, and gastric tumors. PMID- 8405880 TI - Prostaglandins as mediators of lower esophageal sphincter dysfunction in esophagitis. PMID- 8405881 TI - Platelet-activating factor antagonists and infected cirrhosis. PMID- 8405882 TI - Systemic lupus erythematosus and pancreatitis. PMID- 8405883 TI - Ontogeny of Ia messenger RNA in the mouse small intestinal epithelium is modulated by age of weaning and diet. AB - BACKGROUND: Exogenous antigenic peptides are presented to T cells by class II major histocompatibility complex (Ia) molecules on the surface of antigen presenting cells. Class II-associated invariant chain (Ii) is also required for effective antigen presentation. Because messenger RNAs (mRNAs) for Ii chain and for class II I-A beta chain appear in the mouse intestinal epithelium after weaning, experiments were conducted to test the effect of age of weaning and diet on the appearance of Ia and Ii mRNA. METHODS: Four litters were split at day 17; one half was weaned and the other remained with the mother until day 24. On day 23, 25, 27, and 29, enterocytes were isolated from full-length small intestine by vascular perfusion with 30 mmol/L ethylenediaminetetraacetic acid, and the RNA was extracted. RESULTS: Appearance of Ii and I-A beta was significantly delayed by late weaning, as judged by RNA hybridization blots (Ii chain) and complementary DNA amplification (I-A beta chain). In mice on elemental diets, the appearance of Ii and I-A beta chain was delayed compared with littermates reared on standard chow. Ii mRNA failed to appear in mice maintained on the elemental diet by day 40, despite normal growth. CONCLUSIONS: Appearance of mRNA for both Ia and Ii depends on the introduction of a complex diet and not the "stress" of weaning or elimination of breast milk. Introduction of foreign dietary antigens or development of an altered intestinal flora may contribute to this process. PMID- 8405884 TI - Fluid loading of the human colon: effects on segmental transit and stool composition. AB - BACKGROUND: Because segmental storage by the colon is relevant to diarrheal states, we quantified colonic transit in 13 healthy volunteers. METHODS: We infused radiolabelled liquids and solids into the cecum. Gamma camera images were obtained for 48 hours and counts measured in ascending (AC), transverse (TC), descending (DC), and rectosigmoid (RS) colons. Bowel movements were scored for consistency and stool outputs of isotopes were quantified. RESULTS: No volunteers developed diarrhea. Times for isotopes to transverse AC and TC were shorter with rapid infusions (P < 0.001). Counts remaining in AC and TC were also correlated with infusion volumes for 9 hours but not thereafter. Solids were initially delayed in AC but solids and liquids were stored equally in TC. Transit through DC was rapid, but RS stored contents for many hours. First appearance of counts in stools and total counts excreted after 48 hours were not dependent on the infusion rate. Fecal consistency and water content correlated significantly with colonic transit times. CONCLUSIONS: Fluids moved ahead of solids in the AC, but liquids and solids were stored equally in the TC. The DC acted mainly as a conduit during fluid overload but the RS stored both isotopes extensively. Stool consistency was a valid reflection of total colonic transit. PMID- 8405885 TI - The cardiovascular and renal effects of hydrins and arginine vasotocin in frogs. AB - Cardiovascular and renal actions of hydrins and arginine vasotocin (AVT) in frogs (Rana tigrina and Rana catesbeiana) were examined in view of their relationships as precursor/prohormone-hormones in the anuran amphibian. In vitro hydrins and AVT respectively vasorelaxed and vasoconstricted KCl-preconstricted femoral artery in R. tigrina. AVT also produced dose-dependent contractions on untreated preparations; hydrins had no effect on these preparations. Both the hydrins and AVT were positively inotropic and chronotropic, with AVT being more potent. In vivo hydrins and AVT were anti-diuretic except at high doses, when AVT became diuretic. The hydrins and AVT were vasodepressor and pressor, respectively, in R. catesbeiana. These data suggest that the addition of one to three amino acids onto the AVT molecule reverses the contractile response in the vascular smooth muscle. With this dissociation of the vascular effect from that of the renal, the hydrins might play a physiological role in water balance of frogs. PMID- 8405886 TI - At low dose, arginine vasotocin has vasopressor rather than vasodepressor effect in chickens. AB - It is widely accepted that both arginine vasopressin (AVP), and its avian analogue, arginine vasotocin (AVT) are vasopressor (VP) in mammals, but vasodepressor (VDP) in birds. The rat VP and chicken VDP assays were used to characterize neurohypophysial peptides and their analogues. However, these assays use pharmacological doses. Like AVP in mammals, AVT is secreted in birds during dehydration, when peripheral vasoconstriction rather than vasodilatation is needed. Mature White Leghorn cockerels with permanent cannulas in brachial artery and vein were put restrained in a sling. Mean arterial pressure (MAP); heart rate; and rectal, shank, skin, and comb temperatures were continuously recorded. The i.v. infusion regimen was: 1 hr of 0.05 ml/kg*min of saline followed by an hour of tested drug and another hour of saline. The drugs were: AVT, V1-agonist ([Phe2 Orn8]VT), and V2-agonist (DDAVP), in dosage of 50 pmol/kg*min each. AVT and to a lesser extent the V1-agonist increased MAP, caused bradycardia, and reduced thermal conductivity between core and skin or comb. The V2-agonist had no significant effect. Thus, at a low dosage AVT has a VP rather than a VDP effect. Bolus i.v. injection of 2 nmol AVT/kg caused an immediate and large drop in MAP accompanied by tachycardia which subsided within 20-30 sec. This was followed by a small rise in MAP and significant bradycardia. Bolus injection of 2 nmol/kg of either agonist had no hypotensive effect and only the V1-agonist caused the second-phase bradycardia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405887 TI - Cloning and analysis of the gene encoding hummingbird proinsulin. AB - Because hummingbirds exhibit the highest mass-specific metabolic rates seen among vertebrates and rely on sugars as their main energy source, we have investigated the structure of hummingbird insulin (Selaphorus rufus) to determine whether it possesses structural adaptations that increase its receptor binding affinity (potency). We report here the nucleotide sequence of hummingbird proinsulin determined from hummingbird genomic DNA. The predicted amino acid sequence of the A-chain of hummingbird insulin is identical to that of chickens and the B-chain differs by only one amino acid at a noncritical position, B2 (Val in hummingbird and Ala in chicken). These findings suggest that alterations in secretory and metabolic dynamics of insulin are of greater importance than changes in binding affinity in the adaptation to states of high carbohydrate flux in these very energetic organisms. PMID- 8405888 TI - The presence of atrial natriuretic peptide (ANP) in spermatozoa of Bufo arenarum, and effects of GnRH. AB - Atrial natriuretic peptide (ANP)-like immunoreactivity has been demonstrated in the testes of adult Bufo arenarum. Reaction was limited to the head of mature spermatozoa attached to Sertoli cells, interstitial tissue, and seminiferous tubules walls. An immunohistochemical study of isolated spermatozoa showed the reaction to be limited to the basal area of spermatozoon head. After spermiation induced by GnRH, testicular spermatozoa were not immunoreactive. Neither isolated spermatozoa from homogenized testes nor isolated spermatozoa from urine immunostained. The demonstration of immunoreactive ANP in B. arenarum testes, its specific location in specific regions of spermatozoa, and the absence of immunostaining after GnRH treatment, suggest that the "cardiac" peptide may be involved in testicular function. PMID- 8405889 TI - Acceleration of anuran amphibian metamorphosis by corticotropin-releasing hormone like peptides. AB - Despite substantial information on the role of the pituitary-thyroid and pituitary-interrenal axes in controlling amphibian metamorphosis, the hypothalamic hormones responsible for controlling the activity of these axes have not been identified. The mammalian thyrotropin-releasing hormone (TRH) does not regulate the thyroid axis of tadpoles; however, corticotropin releasing hormone (CRH) stimulates the release of thyrotropin from bullfrog tadpole pituitary glands in vitro and may thus function as a central regulator of the thyroid axis during metamorphosis. I tested the possibility that a CRH-like peptide is involved in controlling amphibian development by treating tadpoles of two anuran species, the western spadefoot toad Scaphiopus hammondii, and the North American bullfrog, Rana catesbeiana, with neuropeptides and monitoring their effects on metamorphosis. Injection of spadefoot toad tadpoles with ovine (o) CRH (2 micrograms/animal every other day for 3 weeks) or the amphibian CRH-like peptide sauvagine (SV) significantly decreased their time from hatching to metamorphic climax (Gosner stage 42; frontlimb emergence) and their body weight and body length at climax compared with vehicle-injected controls; whereas, TRH had no effect and arginine vasotocin produced a small but significant lengthening of the larval period but did not alter body size at climax. In an acute response experiment, S. hammondii tadpoles (in Gosner stages 36-38--late prometamorphosis) treated with oCRH or SV (2 micrograms/animal) exhibited significantly elevated whole-body thyroxine (T4) content at 2 and 6 hr after injection; whereas, treatment with TRH (2 micrograms/animal) did not significantly alter whole-body T4. R. catesbeiana tadpoles treated with oCRH or SV (surgical implantation of ELVAX pellets impregnated with 100 micrograms peptide and injections of peptides at 5 micrograms/animal once every 3 days) exhibited accelerated spontaneous and triiodothyronine (T3)-induced metamorphosis as assessed by changes in tail height, hind limb development, and body weight; TRH had no effect. Injections of a pool of antisera generated against CRH-like peptides (rat/human CRH, oCRH, SV) slowed T3-induced metamorphosis when compared with normal serum-injected controls. These results support the hypothesis that a CRH-like peptide(s) is involved in the central control of metamorphosis of anuran amphibians, and may act, at least in part, through stimulation of the thyroid axis. PMID- 8405890 TI - Increase in ovarian alpha-inhibin gene expression and plasma immunoreactive inhibin level is correlated with a decrease in ovulation rate in the domestic hen. AB - This study was designed to examine the relationship among ovulation rate, inhibin alpha-subunit gene expression, and plasma immunoreactive inhibin levels between hens laying at a high ovulation rate and hens laying at a low ovulation rate. A porcine inhibin alpha-subunit cDNA was used to examine the expression of inhibin alpha-subunit in the granulosa cell layers of preovulatory chicken follicles. Total RNA was extracted from granulosa layers of hens laying at a high and low rate and used in slot blot analyses. A dose-related increase in signal intensity with increasing doses of RNA for hens laying at a high and low rate was observed. The expression of the alpha-subunit of inhibin (per microgram of RNA) in granulosa layers of hens laying at a low rate was found to be greater than that in hens laying at a high rate (P < 0.001). Different groups of hens laying at the same rates as those used in the RNA determinations were selected for determination of plasma inhibin levels. Plasma samples from those hens laying at a high and low rate were collected from 1000 to 1800 hr at 2-hr intervals. Plasma inhibin, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were determined by RIA. Plasma inhibin level was higher in hens laying at a low rate than that in those laying at a high rate (P < 0.05), while plasma LH level in hens laying at a low rate was lower than that in hens with a high ovulation rate (P < 0.01). There was no difference in plasma FSH level associated with rate of lay. Results in this study suggest that the ovulation rate-related changes in inhibin alpha-subunit gene expression and plasma immunoreactive inhibin level do not directly affect ovulation rate through a modulation of plasma FSH level. PMID- 8405891 TI - Effects of natural, synthetic, aromatizable, and nonaromatizable androgens in inducing male sex differentiation in genotypic female chinook salmon (Oncorhynchus tshawytscha). AB - The relative potency of several androgens to induce the male phenotype in sexually undifferentiated genotypic female chinook salmon were compared in two separate experiments. The aromatizable and nonaromatizable androgens testosterone (T) and 11-ketotestosterone (11-KT), and the synthetic aromatizable and nonaromatizable androgens 17 alpha-methyltestosterone (MT) and 17 alpha methyldihydrotestosterone (MDHT) were administered to newly hatched alevins in a single 2-hr immersion treatment at doses ranging from 3.2 micrograms/liter to 10 mg/liter. The influence of these treatments on sex differentiation was evaluated by the histological examination of the resulting gonads 6 and 11 months later. In the control group, which was not exposed to exogenous steroids, no males or intersex fish were observed. In contrast, essentially 100% masculinization occurred in groups exposed to MDHT at dosages of 400 micrograms/liter and higher. Treatment with the aromatizable androgen MT resulted in a dose-dependent masculinization, with the production of 100% males at 400 micrograms/liter. However, higher doses resulted in fewer males. 11-KT and T were less potent than the synthetic androgens. The number of males produced after treatment with 11-KT followed a dose-dependent pattern while T showed virtually no masculinizing effect in inducing male phenotype in these studies. The resultant AD50 dosage levels (dosage at which 50% of the genotypic females were sex-reversed into phenotypic males) after a single 2-hr immersion treatment were: 30, 60, and 500 micrograms/liter for MDHT, MT, and 11-KT, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405892 TI - Thyroxine-induced changes in metabolic rate and cytochrome oxidase activity in Thamnophis sirtalis: effects of nutritional status. AB - The effects of nutritional status on thyroxine-induced changes in standard metabolic rate (SMR), and the activity of hepatic cytochrome oxidase were examined in the garter snake Thamnophis sirtalis. Twelve snakes were fed ad libitum, and 12 more were fed a maintenance diet, which was half as many fish per gram of body weight as that eaten by ad libitum snakes. Snakes in the first group gained weight during the 3-week treatment, while individual snakes in the second group either maintained their original weight or showed a slight loss (less than 10%). Within each diet treatment, half of the snakes received a 5-mg thyroxine (T4) pellet implant, and half received placebo implants. Plasma [T4] was unchanged by treatment. Plasma [T3] was elevated in T4-supplemented snakes fed ad libitum, but the difference was not statistically significant (P = 0.09). Standard metabolic rate and cytochrome oxidase activity at 25 degrees were increased significantly (34 and 24%, respectively) only in the T4-supplemented snakes on the ad libitum diet. Thus, T. sirtalis must be in a positive energy balance for thyroid hormones to have an effect on SMR or hepatic cytochrome oxidase activity. PMID- 8405893 TI - Does salmon brain produce insulin? AB - To address the question whether fish brain can produce insulin, pink salmon (Oncorhynchus gorbusha) brains were extracted and processed according to the procedure developed for purification of pancreatic insulin (Rusakov and Bondareva, 1979). Biological and immunological activity of the resulting material was evaluated respectively by a cartilage sulfation assay and by radioimmunoassay homologous for salmon insulin. Preparations from salmon brain stimulated the [35S]sulfate uptake into salmon branchial cartilage with a potency comparable to pure mammalian or salmon insulins but lower than that of mammalian insulin-like growth factor (IGF-I). In contrast, only trace amounts of radioimmunoreactive insulin could be detected by homologous radioimmunoassay. To determine whether insulin mRNA was present in salmon brain, primers specific for salmon proinsulin and salmon prepro-IGF-I were designed to amplify corresponding cDNA regions by reverse transcriptase-PCR. Insulin mRNA was found only in the endocrine pancreas (Brockmann body) while IGF-I mRNA was detected in the brain, liver, and the Brockmann body. Our results suggest that in fish pancreatic-type insulin is most likely produced only in the endocrine pancreas and then transported to the brain through blood/cerebrospinal fluid system. However, it does not exclude a possibility that some yet unknown insulin-like substances may be expressed in the neural system of ectotherm vertebrates. PMID- 8405894 TI - Regulation of testicular steroid production in vitro by gonadotropins (GTH I and GTH II) and cyclic AMP in coho salmon (Oncorhynchus kisutch). AB - The ability of coho salmon gonadotropins GTH I and GTH II to stimulate testicular production of 11-ketotestosterone (11-KT) and 17 alpha,20 beta-dihydroxy-4- pregnen-3-one (17,20 beta-P) in vitro, as well as the involvement of cyclic AMP (cAMP) as a mediator of the actions of GTH I and GTH II, were investigated in maturing male coho salmon. Testicular tissue was incubated in the presence or absence of the test substances at 15 degrees. Both GTH I and GTH II stimulated the in vitro production of 11-KT and 17,20 beta-P in a concentration- and time dependent manner. In testicular tissue at stage IV of spermatogenesis, GTH I and GTH II stimulated the in vitro production of steroids with similar potency. The steroidogenic activity of GTH I and GTH II appeared to be mediated, at least in part, by cAMP as indicated by the ability of dibutyryl cAMP and forskolin to mimic the steroidogenic actions of GTHs, and the similar ability of GTH I and GTH II to increase intracellular cAMP levels in the testicular tissue. The calcium ionophore A23187 stimulated basal but not GTH-stimulated production of steroids in vitro. In addition, differential effects of a protein kinase C inhibitor, H-7, on 11-KT and 17,20 beta-P production were found. Basal and GTH-stimulated production of 11-KT were inhibited by H-7; whereas basal and GTH-stimulated production of 17,20 beta-P were stimulated by H-7. These results suggest that protein kinase C may also be involved in the steroidogenic actions of GTH I and GTH II. PMID- 8405895 TI - Immunolocalization of androgen receptor in the small, preovulatory, and postovulatory follicles of laying hens. AB - Previous studies have indicated that androgens may act directly on the ovarian follicles to regulate their functions. The aim of this study was to localize androgen receptor (AR) in the small, preovulatory and postovulatory follicles of laying hens by an immunocytochemical method and Western blot analysis. Small follicles embedded in the stroma (SF), small white follicles protruding from the surface of ovary (SWF), the third largest (F3) and largest follicles (F1), and the most recent postovulatory follicle (POF) were obtained from hens approximately 4 or 10 hr before the expected time of ovulation. Frozen sections of these follicles were immunostained by using anti-human AR antibody. Furthermore, the granulosa cells and theca tissue of preovulatory follicles (F1 and F2) approximately 4 hr before the expected time of ovulation were processed for western blot analysis for AR. The majority of granulosa cell of SF showed negligible AR immunoreaction, whereas all of the granulosa cells of SWF, F3, F1, and POF exhibited a strong AR immunoreaction. Thecal interstitial cells in SWF, F3, and F1 stained positive for AR, and those in POF showed only a weak immunoreaction. The thecal fibroblasts of SWF, F3, and F1 also showed a positive AR immunoreaction, whereas those of POF stained weakly. No significant difference in the AR localization in the ovary was observed between 4 and 10 hr before the expected time of ovulation. Western blot analysis indicated that the granulosa cells and thecal tissue contained AR protein of molecular weight of approximately 120,000.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8405896 TI - The primary structure of sturgeon prolactin: phylogenetic implication. AB - The complete amino acid sequence of prolactin (PRL) from a chondrostean species, the sturgeon (Acipenser gueldenstaedti), has been determined. Sturgeon PRL was isolated from the pituitary glands by gel filtration on a Sephadex G-25 column and high-performance liquid chromatography on a reverse-phase column following acid-acetone extraction. Sturgeon PRL was identified by immunoblot reactivity using antisera against salmon and ovine PRL. It consists of 204 amino acid residues, which is the largest among known PRLs, and contains three disulfide bonds corresponding to those of tetrapod PRLs. Sequence comparison with PRLs from other vertebrates revealed that sturgeon PRL has slightly higher sequence identities (35-46%) with teleost PRLs than with tetrapod PRLs (30-40%). These structural characteristics imply that an ancestor of the ray-finned fishes had PRL with three disulfide bonds and at some point after divergence of Chondrostei, the disulfide bond in the amino-terminus of PRL was lost. PMID- 8405897 TI - Primary structures of glucagon and glucagon-like peptide isolated from the intestine of the parasitic phase lamprey Petromyzon marinus. AB - Immunohistochemical studies have indicated that glucagon-containing cells are present in the intestinal mucosa of the agnathan Petromyzon marinus (sea lamprey) but are absent from the pancreas. Glucagon was isolated from an extract of intestinal tissue taken from specimens of sea lamprey during their parasitic phase. The primary structure of the peptide was established as: His-Ser-Glu-Gly Thr5-Phe-Thr-Ser-Asp-Tyr10-Ser-Lys-Tyr-Leu- Glu15-Asn-Lys-Gln-Ala-Lys20-Asp-Phe Val-Arg-Trp25-Leu- Met-Asn-Ala. This amino acid sequence shows 8 substitutions compared with that of mammalian glucagon but, with the exception of the COOH terminal alanine residue, lamprey gut glucagon contains no structural features that have not been previously seen in glucagons isolated from the pancreata of gnathostomes. The amino acid sequence of lamprey glucagon-like peptide (GLP) demonstrates that the primary structure of this peptide has been less well conserved than that of glucagon. The sequence His-Ala-Asp-Gly-Thr5-Phe-Thr-Asn Asp-Met10-Thr-Ser-Tyr- Leu-Asp15-Ala-Lys-Ala-Ala-Arg20-Asp-Phe-Val-Ser-Trp25- Leu Ala-Arg-Ser-Asp30- Lys-Ser shows 16 amino acid substitutions compared with the corresponding region of mammalian GLP-1 and 15 substitutions compared with that of salmon GLP. PMID- 8405898 TI - Isolation of gonadotropin subunits and evidence for two distinct gonadotropins in Atlantic croaker (Micropogonias undulatus). AB - Gonadotropic hormones (GTHs) were purified from Atlantic croaker (Micropogonias undulatus) pituitary glands collected in December, when the gonads were regressed. GTH was identified during the purification by an in vitro ovarian steroidogenesis bioassay and by radioimmunoassay using an Atlantic croaker maturational GTH antiserum. Two gonadotropic fractions were separated by anion exchange chromatography of gel-filtered pituitary extracts. Reverse-phase HPLC followed by n-terminal amino acid sequencing of the GTH subunits revealed that a common alpha-subunit was present in both fractions. However, the two gonadotropic fractions contained different forms of the beta-subunit which differed both in their n-terminal amino acid sequences and in their amino acid compositions. The differences in the amino acid compositions between the two croaker GTH beta subunits were similar to those previously reported between salmon GTH I and II beta-subunits (Kawauchi et al., 1989, Fish Physiol. Biochem. 7, 29-38). The n terminal sequence of one of the croaker beta-subunits (beta-II) resembled that of the salmon GTH II beta-subunit. The other subunit (beta-I) had only a low degree of homology in its n-terminal sequence with the salmon GTH I beta-subunit but had a similar amino acid composition. In addition, the n-terminal end of beta-I had 40% sequence identity with the GTH I beta-subunit of bonito (Kawauchi et al., 1991, In "Proceedings of the Fourth International Symposium on Reproductive Physiology of Fish (A. P. Scott, J. P. Sumpter, D. E. Kime, and M. S. Rolfe, Eds.), pp. 19-21. Univ. of East Anglia, Norwich, UK). Moreover, the beta-I subunits from all three species had cysteine residues at positions 4 and 8. These results indicate that a dual gonadotropin system exists in the Atlantic croaker as in the salmon and bonito and support the concept that such a system is ubiquitous in teleosts. PMID- 8405899 TI - Population declines in the snowshoe hare and the role of stress. AB - Every 10 years snowshoe hare populations across the boreal forest of North America go through a population cycle, culminating in a decline lasting 4 or more years. We tested the hypothesis that snowshoe hares during the decline are in poor condition and less able to respond to challenges in their environment by examining the stress response of male hares. Three groups from February and May, 1991 (the second year of the hare decline in the Yukon), were compared: baseline hares were collected to obtain resting hormone levels; control hares were wild animals caught at randomly placed sites; and fed hares were wild animals caught on supplementary fed areas. The latter two groups were sequentially bled to examine their response to dexamethasone (DEX) followed by adrenocorticotropic hormone (ACTH). Trapping and handling were stressful to the experimental hares as the initial blood levels of total and free cortisol levels were higher (especially in controls), testosterone levels were lower, and glucose levels were higher in experimental hares than in baseline hares. Control and fed hares showed similar total and free cortisol responses, falling to low levels after the DEX injection and increasing rapidly in response to the ACTH injection. However, control hares were in worse condition than fed hares as indicated by the higher free cortisol levels and lower maximum corticosteroid-binding capacity (MCBC) in control hares. In addition, though testosterone levels fell in both groups in response to DEX, only the fed hares showed a large, transitory increase 30 min after the ACTH injection. An unexpected finding was a dramatic increase in MCBC levels 30 min after the ACTH injection in both experimental groups, but it was more pronounced in the fed group. We conclude that the pituitary-adrenocortical feedback system in hares from declining populations is operating normally and that they should be able to cope with acute, short-term stressors, but that they are in poor condition and are exposed to higher levels of free cortisol than fed hares in good condition. PMID- 8405900 TI - Sex change and steroid profiles in the protandrous anemonefish Amphiprion melanopus (Pomacentridae, Teleostei). AB - Plasma profiles of several gonadal steroids and cortisol were examined in a field population of the protandrous, sex-changing anemonefish Amphiprion melanopus to elucidate potential roles of these hormones in gonadal sex change. Sex change was experimentally induced in males by removal of their dominant female pair mates. These sex-changing males were captured and sampled at 5, 10, or 20 days after female removal. Unmanipulated males and females were also sampled. Males had higher plasma levels of 11-ketotestosterone (11-KT) than did females, but had lower levels of androstenedione (Ad), testosterone (T), and estradiol-17 beta (E2). The three androgens showed decreases from male levels at 10 days, then an increasing trend at 20 days after female removal. E2 levels exhibited no changes from male levels until 20 days, when a significant increase over male levels was observed. Mature females had higher levels of Ad, T, and E2 than the 20-day treatment group, indicating that these steroids continue to rise after Day 20. The results support hypothesized roles for androgens in male function and E2 in female function in A. melanopus. However, E2 increases lagged behind oogonial proliferation, arguing against an influence of this steroid in the initiation of female function. Cortisol levels did not differ between males and females, but exhibited an increase during sex change, peaking at 20 days. PMID- 8405901 TI - Calbindin D28K regulation in precociously matured chick egg shell gland in vitro. AB - Egg shell calcification in the hen uterus (egg shell gland, ESG) depends primarily on intestinal absorption of dietary Ca2+ as well as ESG Ca2+ transport into the shell. Intestinal Ca2+ absorption is linked to vitamin D-induced calbindin D28K (D28K) concentration. The ESG also contains D28K, and Ca2+ transport into the shell appears to be linked to D28K gene expression, but until this report, there was no direct proof that ESG D28K was or was not vitamin D dependent. To address this issue, highly developed ESG from estradiol (E2) injected, severely vitamin D-depleted chicks were cultured in serum-free medium with excellent viability. Addition of the vitamin D-hormone, 1,25(OH)2 vitamin D3 (1,25), to the culture medium increased ESG D28K levels as much as 70%. E2 alone had no effect, but E2 plus 1,25 further increased ESG D28K levels up to 160%. By contrast, progesterone (P4) prevented the 1,25-stimulated increase in D28K, while having no effect on basal D28K level. Of considerable interest, thapsigargin (THAPS), which increases intracellular Ca2+ concentration ([Ca2+]i) in many cell types, stimulated D28K synthesis in a concentration-dependent manner in the complete absence of 1,25 and independent of the [Ca2+] of the medium. These results are the first direct evidence that ESG D28K is under direct control of 1,25 and that both gonadal steroid hormones, E2 and P4, may be coregulators. Further, the effects of THAPS suggest that [Ca2+]i itself may also regulate D28K. This new in vitro model clearly represents a unique opportunity to study the regulation of the ESG calcium transport mechanism under stringently defined conditions. PMID- 8405902 TI - Immunocytochemical localization of serotonin and gonadotropin-releasing hormone in the brain and pituitary gland of the Atlantic croaker Micropogonias undulatus. AB - The distribution of serotonin-immunoreactive (5-HT-ir) and gonadotropin-releasing hormone-immunoreactive (GnRH-ir) elements in the brain and pituitary gland of the Atlantic croaker (Micropogonias undulatus) was studied using immunoperoxidase and immunofluorescence techniques. 5-HT- and GnRH-ir perikarya were observed in the ventrolateral region of the olfactory bulbs and in the preoptic-anterior hypothalamic area bordering the third ventricle as well as in regions distal to the ependyma of the third ventricle. The immunoreactive elements of the two systems appear to overlap in these regions. Cerebrospinal fluid-contacting 5-HT ir neurons were found in the periventricular hypothalamus, particularly in the preoptic area and in the nucleus posterior periventricularis, nucleus ventromedialis thalami, nucleus recessus lateralis, and nucleus recessus posterioris. Immunoreactivity to 5-HT in the pituitary was strongest in the proximal pars distalis (PPD) and in a few scattered cells bordering the pars intermedia. Dendritic processes of 5-HT neurons could be seen contacting the adjoining hypophysial cells in the PPD. GnRH-ir fibers were seen entering the pituitary stalk and the immunoreaction was concentrated in the ventral PPD. Several large 5-HT-ir perikarya were located in the raphe region forming a column in the mediosagittal plane. The results suggest that the general organization of the 5-HT system in the brain of the Atlantic croaker is similar to that described in other vertebrates. The demonstration that 5-HT and GnRH systems are in close proximity, particularly in the olfactory bulbs and in the preoptic-anterior hypothalamic area of the croaker brain, suggests that the two systems may interact in these regions of the teleostean brain to control several reproductive functions including spawning behavior and gonadotropin release. PMID- 8405903 TI - Pregnancy-associated factors affecting organic phosphate levels and oxygen affinity of garter snake red cells. AB - Pregnancy in Thamnophis elegans is associated with an increase in the nucleoside triphosphate (NTP) concentration and a concomitant decrease in the oxygen affinity of the adult red cell. Red cell NTP levels rise at about the time of ovulation and peak during mid-gestation. Since plasma progesterone levels appear to have a similar profile, we examined the influence of progesterone on this phenomena. Surgical removal of the corpora lutea (CL), primary site of progesterone release, plus fetuses abolished the pregnancy-associated effect on red cell NTP levels. Removal of the CL alone resulted in NTP levels intermediate to those of the red cells of the nonpregnant and the pregnant groups. Progesterone implants in nonpregnant females, as well as in males, caused red cell NTP concentrations to rise. These data support the hypothesis that progesterone, secreted by the CL, in the presence of the fetus, is largely responsible for the pregnancy-associated increase in the red cell NTP concentration. PMID- 8405904 TI - Inhibitory effects of n-alkanols on the hormonal induction of maturation in follicle-enclosed Xenopus oocytes: implications for gap junctional transport of maturation-inducing steroid. AB - In fishes and amphibians, the induction of oocyte maturation by gonadotropin is mediated by follicle cell production of maturation-inducing steroid. This steroid appears to be progesterone in the frog Xenopus laevis. It is also known that full grown oocytes of X. laevis are cytoplasmically linked to follicle (granulosa) cells via gap junctions, and that these gap junctions are enhanced by whole animal injections of gonadotropin (human chorionic gonadotropin, hCG). However, the specific role of this heterocellular gap junctional communication is unclear, especially in regard to its possible involvement in the regulation of oocyte maturation. Therefore, the objective of this study is to examine the role of follicle cell-oocyte gap junctional coupling during maturation of the ovarian follicle of X. laevis. For this purpose, we determined the in vitro effects on oocyte maturation of the known gap junction uncouplers 1-heptanol and 1-octanol, alone or in combination with maturational concentrations of hCG or progesterone. The n-alkanols alone did not have any effect on maturation. However, both heptanol and octanol at 1 mM blocked the hCG and progesterone induction of maturation in follicle-enclosed Xenopus oocytes. At least in the case of heptanol, the inhibition of hormone-induced maturation was not accompanied by changes in the steroidogenic response of intact follicles to hCG or in the responsiveness of defolliculated oocytes to exogenous progesterone. These latter observations seem to rule out non-specific effects of the alcohols on follicle cell or oocyte function. Therefore, our results suggest that gap junctional coupling between follicle cells and the oocyte is required for maturational steroid signaling in amphibian ovaries. PMID- 8405905 TI - In vitro steroid secretion during early development of mono-sex rainbow trout: sex differences, onset of pituitary control, and effects of dietary steroid treatment. AB - Sex differentiation in many teleost species can be controlled by treatment with steroids. To investigate the development of steroidogenesis during both natural and controlled sex differentiation, the production of androstenedione, testosterone, and estradiol were determined in tissues from populations of all female and all-male rainbow trout (Oncorhynchus mykiss). At various times from hatching through gonadal sex differentiation, explants of steroidogenic tissues were incubated in vitro alone or in the presence of partly purified salmon gonadotropin and the resulting media were assayed for steroids. Androstenedione and testosterone were produced at higher levels in media from testes than from ovaries within 2 weeks of the onset of feeding (before any dramatic gonadal differentiation). Gonadal estradiol secretion was nondetectable until about 1 month after the onset of feeding when females produced more than males. Gonadotropin stimulated gonadal steroid production only after differentiation, but stimulated anterior kidney (interrenal) production of androstenedione much earlier in development. Dietary treatment of rainbow trout with either estradiol or 17 alpha-methyltestosterone (MT) inhibited in vitro gonadal steroid production and this effect persisted in MT-fed fish even after withdrawal of dietary steroids. PMID- 8405906 TI - Breeding cycles and fecal gonadal steroids in the brown dipper Cinclus pallasii. AB - Breeding cycles and fecal gonadal steroids were investigated in free-living brown dippers, Cinclus pallasii. The brown dippers were marked with colored leg bands, and their behavior was observed by a field scope. The breeding season was divided into six stages according to the behavior observed; winter territory-defending, nest-building, copulation, incubation, nestling, and fledgling stages. Fecal pellets on the stones in their territories were collected and the levels of immunoreactive estradiol-17 beta (E2) and testosterone (T) in 10 mg feces were measured by radioimmunoassay. Levels of fecal E2 were high only during the copulation stage, which lasted for a few weeks, in all females of the three pairs observed. In contrast, T levels in the feces of males increased in late winter when intensive territory-defending behavior was observed, and remained high until the end of the breeding season. T levels during the nest-building stage and the copulation stage were similar to those during other nesting stages. When the birds breed only once a year, the duration of high levels of T was shorter than that of the pair with the second clutch. T levels in the feces of females showed a similar pattern to E2 levels. PMID- 8405907 TI - Calcium-permeable channels in HeLa cells. AB - Calcium-permeable channels with a slope conductance of 9 pS were revealed in excised inside-out patches of cultured HeLa cells. Over a potential range from 80 to -10 mV, unitary inward currents were recorded with 110 Ca2+ in the pipette in artificial "intracellular" solutions containing impermeant anions. Channel activity was not considerably affected by varying free calcium concentration between 0.01 and 10 mumol/l in cytosol-like solution. In experiments with low spontaneous activity of calcium-permeable channels in the inside-out patch, it could be increased by the application of 5-10 mumol/l inositol-trisphosphate to the inner membrane surface. PMID- 8405908 TI - Effect of adenosine 5'-triphosphate on secretagogue-stimulated (14C)-aminopyrine accumulation by rabbit isolated gastric glands. AB - The effect of adenosine 5'-triphosphate (ATP) on gastric acid secretion stimulated by histamine, carbachol and dibutyryl-cAMP (db-cAMP) was studied using glands isolated from rabbit gastric mucosa. The (14C)-aminopyrine (AP) accumulation method was used as an index of acid production by the gastric glands. Histamine-stimulated AP accumulation was significantly inhibited by ATP (10 mumol/l-1 mmol/l). The inhibitory action of ATP appeared to be specific, inasmuch as this nucleotide had no significant effect on basal secretion or secretion stimulated by carbachol or db-cAMP. The antisecretory effect of ATP on histamine-stimulated glands was not affected by the P1-purinoceptor antagonist, theophylline. Pretreatment of glands with indomethacin, a well known prostaglandin synthesis inhibitor, led to a significant reduction of the inhibitory responses to ATP. These results show that ATP inhibits the histamine stimulated AP accumulation by rabbit isolated gastric glands and suggest that this effect is not due to an ectoenzymatic conversion of ATP into adenosine but to a direct effect of ATP which may be mediated via a P2-purinoceptor subtype linked to prostaglandin production. PMID- 8405909 TI - Kinetic analysis of the L-type calcium current in enzymatically dissociated ferret ventricular myocytes. AB - The L-type calcium current (ICa-L) was studied in single ferret ventricular myocytes using whole-cell recording with single patch pipettes. Voltage-clamp experiments were performed at room temperature with internal and external Na(+)- and K(+)-free Tyrode solutions in order to isolate ICa-L. For depolarizing steps eliciting small ICa-L the decay of the current is best described by one exponential. For depolarizing steps eliciting large ICa-L (i.e. between -10 and +30 mV), the decay of the current is best described by the sum of two exponentials with a calcium-dependent fast (Tf) time constant and a voltage dependent slow (Ts) time constant. Experiments conducted with different external concentration of Ca2+ and Ba2+ suggested that the inactivation and the time course of reactivation of the current after a depolarizing pulse are dependent on calcium ions. This confirms previous observations in heart muscle and reveals the existence of a calcium-dependent regulation process of the L-type calcium current in enzymatically dissociated ventricular myocytes from ferret heart. PMID- 8405910 TI - Irreversible structural changes in thylakoid membranes at high temperatures. Detection by luminescence and EPR. AB - The character of structural changes in thylakoid membranes caused by temperature variation was investigated. Experiments were performed on maize leaf segments in vivo and in a closed temperature cycle 24-50-24 degrees C. Two biophysical methods were used for detection: luminescence and EPR. Arrhenius plots of delayed fluorescence (DF) versus reciprocal temperature revealed two break points, T1 and T2. The MeFASL (10.3) spin probe monitoring properties close to membrane surface detected only T2 transition temperature. The results were interpreted in terms of a fluidity change which starts in a membrane centre at T1 and gradually displaces toward the surface at T2. The T1 and T2 transition temperatures are sensitive to pretreatment history of plants indicating that high temperature and drought induced membrane alterations are irreversible. Activation energies E1, E2 and E3 were determined for temperature regions below T1 between T1 and T2, and above T2, respectively. The E1 and E3 activation energies showed greater sensitivity to stress than did E2. There are some indications that the DF method could be used to screen temperature sensitive and temperature resistant genotypes. PMID- 8405911 TI - An ESR study of manganese binding in plant tissue. AB - Two different fractions of manganese were found in the maize plant root apoplasm (intercellular space containing cell walls) after soaking the roots in MnCl2 solutions (concentration range 0.01-10 mmol.l-1): (a) an Mn2+ fraction in the water free space (WFS) which gave a characteristic six-line spectrum, and (b) an immobilized fraction that gave no detectable ESR spectrum. Both fractions affect proton NMR relaxation (T1) of the tissue water through water exchange across cell membranes. ESR spectra of free and total manganese of the root tissue treated with MnCl2 also revealed different time courses for saturation of WFS and DFS with Mn2+. Binding of manganese in the extracellular space of the tissue seems to be the rate limiting step in permeation of Mn2+ across the root cell membranes. PMID- 8405912 TI - Effect of lipid autoperoxidation on the activity of the sarcoplasmic reticulum (Ca2+-Mg2+)ATPase reconstituted into egg yolk phosphatidylcholine bilayers. AB - The activity of (Ca(2+)-Mg2+)ATPase reconstituted into egg yolk phosphatidylcholine liposomes is reduced when the lipid has been oxidized before the reconstitution procedure. It is suggested that the reversible decrease in activity is caused by a decrease of the lipid bilayer thickness due to the shortening of lipid acyl chains during autoperoxidation; the irreversible decrease in the activity is caused by chemical reactions of the enzyme SH groups with the lipid autoperoxidation products. PMID- 8405913 TI - On the origin of "pure" chirality of amino acids and saccharides at the prebiotical stage. AB - Possible explanation is offered for the origin of pure chirality of amino acids and saccharides of racemic compounds during the prebiotical stage of evolution. It is based on the theory of "noise induced phase transitions", a mechanism approached in previous years by synergetic methods. It follows from this approach that a sufficiently intensive noise in the environment of the formation of racemic compounds of L- and D-chiralic macromolecules could produce homochiralic state, i.e. a state with either pure L- or D-chirality. PMID- 8405914 TI - Influence of polar polymers on the apoprotein region of human serum lipoproteins: an electron paramagnetic resonance (EPR) study. AB - Electron spin resonance spectroscopy was used for measurements of the surface potential and apoprotein structure of LDL and HDL in the presence of Ca2+ and dextran sulfate, heparin and chondroitin sulfate. A decrease in the absolute values of surface potential of LDL and HDL was observed after addition of Ca2+. In the presence of the negatively charged macromolecules the measured surface potential was less reduced. The spectral properties of a maleimide spin label covalently attached to the apoprotein were changed under conditions of aggregation of LDL induced by dextran sulfate, chondroitin sulfate or heparin in the presence of Ca2+. In the HDL system this effect was only observed for dextran sulfate. The influence of PEG on the spectral parameters of the spin label is dependent on the molecular weight of the polymer. PEG 400 decreased the mobility of the spin-labelled apoprotein region of LDL, whereas PEGs with higher molecular weight only slightly increased the maleimide mobility. On the other hand, the maleimide-labelled apoprotein region of HDL showed a higher sensibility to all PEGs used. Addition of PEG leads to immobilization of apoprotein A. PMID- 8405915 TI - Some peculiarities of the Na(+)-Ca2+ exchange reaction in choline, tetramethylenammonium and Tris media. PMID- 8405916 TI - Leu-arylamidase activity levels during the estrous cycle and pregnancy in several extrahypothalamic areas of the rat brain. AB - A wide variety of behavioral changes in the female rat have been associated with the estrous cycle, pregnancy, and the postpartum period and their accompanying hormonal fluctuations. Since aminopeptidase activity, that presumably controls the activity of several neuroactive peptides, has been implicated in the control of these hormonal changes, the present study examined the tissue levels of Leu arylamidase activity (Leu-ArA) in the cortices (frontal, parietal and occipital), striatum, hippocampus, amygdala, pineal gland and medulla oblongata during the estrous cycle, different stages of pregnancy (2nd, 7th, 14th and 20th postinsemination day) and the postpartum day. In the estrous cycle, Leu-ArA was significantly increased during the afternoon of the proestrous in the occipital cortex, amygdala and medulla oblongata. In general, higher enzyme levels were found during the middle stages of pregnancy. There is a significant rise after parturition in the occipital cortex, and clear but insignificant increases in the striatum and medulla. It is suggested that several extrahypothalamic areas may mediate several effects on Gn-RH secretion from the hypothalamus, and that Leu ArA could be implicated. PMID- 8405917 TI - Effects of pertussis toxin on intraocular pressure and adenylate cyclase activity of ciliary processes in rabbits. AB - The effects of pertussis toxin on the intraocular pressure (IOP) lowering effect of clonidine and isoproterenol as well as on the inhibitory effects of clonidine and neuropeptide Y on adenylate cyclase activity of ciliary processes were studied in albino rabbits. I.v. administered pertussis toxin elicited transient changes in IOP which, however, returned to control values during 2-3 days. In the following days the IOP lowering effect of the alpha 2-adrenergic agonist clonidine was abolished and that of the nonselective beta-adrenergic agonist isoproterenol was attenuated. At the same time, the inhibitory effects of clonidine and neuropeptide Y on basal as well as stimulated adenylate cyclase activities in homogenates of ciliary processes were grossly diminished. The effects of pertussis toxin on the IOP lowering action of adrenergic agonists and on the inhibitory action of clonidine and neuropeptide Y on adenylate cyclase activity were ascribed to an impairment of the function of a G protein in ciliary processes, probably G(i) protein. It is suggested that the decrease of IOP induced by clonidine is due to inhibition of adenylate cyclase. PMID- 8405918 TI - Effect of oxidative stress on (3H)N-methylscopolamine binding and production of thiobarbituric acid reactive substances in rat cerebral cortex membranes. AB - We investigated the effect of lipid peroxidation, in vitro induced by H2O2 or FeSO4 and ascorbic acid, on binding properties of muscarinic receptors in rat cerebral cortex membranes. Simultaneously the concentrations of thiobarbituric acid reactive substances (TBARS) were measured to assess the extent of lipid peroxidation. In conditions of increased TBARS levels the density of (3H)N methylscopolamine [(3H)NMS] binding sites in rat cerebral cortex membranes was not affected. Decreased numbers of (3H)NMS binding sites observed in the presence of high concentrations of H2O2 (100 and 1000 mmol.l-1) accompanied by a decrease of TBARS levels might be related to a nonspecific effect of H2O2 on cellular proteins. PMID- 8405919 TI - Interaction of bovine erythrocyte spectrin with aminophospholipid liposomes. AB - Interaction of bovine erythrocyte spectrin with aminophospholipid (phosphatidylethanolamine, phosphatidylserine and their mixture) vesicles was studied by means of intrinsic fluorescence quenching and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene. Similarly as human and pig erythrocyte spectrin, bovine red blood cell spectrin interacts with vesicles prepared from these phospholipids. In model membranes, spectrin induced an increase of order parameter while in natural, red blood cell membranes spectrin binding was rather connected with a decrease in this parameter. The interaction of spectrin with the PE/PS vesicles was not affected by high concentrations of urea. These vesicles also did not protect spectrin from being denatured by urea. PMID- 8405920 TI - High-conductance chloride channels in BC3H1 myoblasts. AB - The existence of a high-conductance voltage sensitive chloride channel in BC3H1 myoblasts is documented. The conductance of the channel in symmetrical 150 mmol/l sodium chloride is around 400 pS. The probability of the channel being in the open state decreases with increasing of the imposed voltage from holding potential (0 mV) in both the depolarizing and the hyperpolarizing direction respectively. The bell-shaped open probability plot is asymmetrical, and can be fitted by two Boltzmann equations with different Vh and kn constants; the fitted values were -53 mV and -8.4 respectively for the negative side and +41 mV and +10.2 for the right side. When the unit Cl currents to rectangular pulses are summated the resulting total ionic Cl current shows relaxation, which increases with the amplitude of the pulse. The activation as well as the shape of the current can be significantly influenced by varying the amplitude and the direction of prepulses or holding potential. The high-conductance Cl channel shows several substrates (at least four with amplitudes of around 100 pS); frequently accompanied by flickerings. PMID- 8405921 TI - The effect of Pb2+ ions on calcium currents and contractility in single muscle fibres of the crayfish. AB - Muscle fibres of crustaceans represent an useful model for studying the mode of action of substances influencing calcium channels as the membrane generates the active responses on a pure calcium principle and the excitation-contraction link is dependent on external calcium. We followed the effect of Pb2+ ions (1-300 mumol/l) on contractile responses in single muscle fibres of the crayfish evoked by massive or intracellular electrical stimulation, by potassium depolarization and caffeine application, as well as on action potential in single intact muscle fibres and on calcium currents in voltage clamp conditions (vaseline gap) in internally perfused muscle fibre segments. All types of contractile responses, single twitches, tetanus, potassium and caffeine contractures were blocked. The strontium action potential was blocked very effectively by Pb2+ ions. The total calcium currents which can be split by means of Hodgkin-Huxley equations into two components (fast and slow respectively) differing in the rate of activation and inactivation were suppressed after addition of Pb(NO3)2 (50-300 mumol/l). The effect of Pb2+ was concentration and time dependent. At lower concentrations (100 mumol/l) the blocking effect was more pronounced on the fast inactivating Ca current component. The Pb2+ ions prolonged the time constant of inactivation tau h of the slow channel, while leaving that of the fast channel intact. PMID- 8405922 TI - Isoprenoid biosynthesis in bacteria: two different pathways? AB - The biosynthesis of isopentenylpyrophosphate, a central intermediate of isoprenoid formation, was investigated in six different bacterial organisms. Cell free extracts of Myxococcus fulvus, Staphylococcus carnosus, Lactobacillus plantarum and Halobacterium cutirubrum converted [14C]acetyl-CoA or [14C]hydroxymethylglutaryl-CoA to [14C]mevalonic acid. Furthermore, [14C]mevalonic acid, [14C]mevalonate-5-phosphate and [14C]mevalonate-5 pyrophosphate were metabolized to [14C]isopentenylpyrophosphate. These data demonstrated the in vitro operation of acetoacetate pathway for the formation of isopentenylpyrophosphate in bacteria. In contrast, no intermediates of this reaction sequence could be detected using cell-free extracts of Zymomonas mobilis and Escherichia coli. These results indicate that at least two different pathways for the biosynthesis of isopentenylpyrophosphate are present in bacteria. PMID- 8405923 TI - Characterization of three different lytic transglycosylases in Escherichia coli. AB - Two lytic transglycosylases, releasing 1,6-anhydromuropeptides from murein sacculi are present in a mutant deleted for the soluble lytic transglycosylase 70 (Slt70). Thus, there are three different lytic transglycosylases in Escherichia coli. One of the remaining enzymes is soluble and one is a membrane protein that can be solubilized by 2% Triton X-100 in 0.5 M NaCl. Both enzymes are exo muramidases. Only the membrane enzyme, but not the soluble ones, hydrolyses isolated murein glycan strands (poly-GlcNAc-MurNAc). While the soluble enzymes are inhibited by the muropeptide TetraTriLysArg(dianhydro), the membrane enzyme is not. The antibiotic bulgecin that inhibits Slt70 does not inhibit the lytic transglycosylases present in the slt70 deletion mutant. PMID- 8405924 TI - Characterisation of a highly repeated DNA sequence from Mycobacterium bovis. AB - We report characterisation of a novel repeat sequence from a Mycobacterium bovis genomic library. The highly repeated sequence belongs to a family consisting of a 24 base pair (bp) direct repeat (DR), that appears to be organized into clusters on the chromosome. We classify the 24-bp DR into the group of prokaryotic DNA repeats known as the interspersed repetitive sequence elements. The 24-bp DR will be of potential use as a DNA fingerprinting tool in epidemiological studies of M. bovis. PMID- 8405925 TI - The existence of a dispensable fibrillar layer on the wall surface of mycelial but not yeast cells of Aureobasidium pullulans. AB - Wall surface ultrastructure of Aureobasidium pullulans was studied by freeze etching. Yeast cells had a smooth wall surface as in typical yeast species. Mycelial cells and chlamydospores had an extra layer on the wall surface made mostly of fibrils. The fibrils were 20 nm in diameter, and thicker than typical major fungal wall skeletal fibrils of beta-glucan and chitin. This layer was apparently easily detached from the wall proper, presumably as a result of enzymic activity or by physical means, suggesting that it is a physiologically dispensable wall component. PMID- 8405926 TI - Retention of a co-translational translocated mutant protein of carboxypeptidase Y of Saccharomyces cerevisiae in endoplasmic reticulum. AB - Co-translational translocation of Saccharomyces cerevisiae vacuolar glycoprotein carboxypeptidase Y (CpY) was highly efficient when studied with an in vivo and in vitro homologous system, comparison of limited proteolytic cleavage of immunoprecipitated translational products of CpY and subcellular localisation of a mutant CpY. The efficient segregation of CpY mRNA in highly purified fractions of rough microsomes was characterised. CpY1 mutant showed retention of core glycosylated material (proCpY1) in the rough and smooth endoplasmic reticulum fractions. It is suggested that the presence of structures that are incompatible with intercompartmental transport of vacuolar protein leads to retention of the mutated CpY by the endoplasmic reticulum. PMID- 8405927 TI - Effect of ethanol on the sterols of the fission yeast Schizosaccharomyces pombe. AB - Ergosterol, lanosterol and two further unidentified sterols were detected and quantified in Schizosaccharomyces pombe cell extracts. In cells grown under anaerobic conditions, the levels of these sterols were dramatically reduced with a concomitant increase of their squalene precursor as compared with cells growing under aerobic conditions. Presence of ethanol resulted in a decrease in the sterol content under aerobic conditions. On the contrary, under anaerobic conditions presence of ethanol resulted in a three-fold increase of total sterols. Lanosterol was the main constituent of this elevation. It is suggested that lanosterol in parallel with unsaturated fatty acids is responsible for maintaining membrane integrity of S. pombe cells growing in the presence of ethanol. PMID- 8405928 TI - Transient repression of the synthesis of OmpF and aspartate transcarbamoylase in Escherichia coli K12 as a response to pollutant stress. AB - The synthesis of total cellular proteins in Escherichia coli K12 was studied in batch culture following exposure of cells to low concentrations of monochlorophenol, pentachlorophenol and cadmium chloride. Changes in protein patterns were identified after pulse-chase labelling of proteins with [35S]methionine and subsequent two-dimensional gel electrophoresis (2D-PAGE). We demonstrated that besides the induction of some stress proteins, also a transient decrease in the rate of synthesis of other proteins occurred. Two of these proteins were identified as OmpF and aspartate transcarbamoylase (ATCase). Their transient repression appeared to be a general response to stress elicited by different pollutants and may therefore be used as a general and sensitive early warning system for pollutant stress. PMID- 8405929 TI - Amino-terminal methionine processing of the protein HPr in Streptococcus salivarius grown in continuous culture. AB - HPr is a protein of the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Streptococci possess two forms of HPr which differ by the presence or the absence of the N-terminal methionine (Met). These forms are called HPr-1 (without Met) and HPr-2 (with Met). In order to determine whether the ratio of these two forms varies with growth conditions, we measured the amount of HPr-1 and HPr-2 present in Streptococcus salivarius grown in continuous culture at pH 7.5. The results indicated that the HPr-1/HPr-2 ratio: 1) was not related to the cellular amount of total HPr; 2) was highest (10.2 +/- 3.5) under glucose (a PTS sugar) limitation (10 mM) and low dilution rate (D = 0.1 h-1; g = 6.9 h); 3) was decreased 2.4- to 5.7-fold when the amount of glucose and/or D was increased; 4) was not influenced by D when cells were cultured on galactose (a non-PTS sugar) but was two-fold higher under conditions of galactose excess (200 mM). We suggest that the cleavage of the N-terminal HPr Met is not a stochastic phenomenon but is dictated by growth conditions. PMID- 8405930 TI - Very efficient extracellular production of cholera toxin B subunit using Bacillus brevis. AB - We have constructed a very efficient synthesis and secretion system for cholera toxin B subunit (CTB) of Vibrio cholerae 569B using Bacillus-brevis. The constructed expression-secretion vector has the multiple promoters and the signal peptide coding region of the mwp gene, a structural gene for one of the major cell wall proteins of B. brevis strain 47, directly followed by the gene encoding the mature CTB. A large amount of mature CTB (1.4 g per liter of culture) was secreted into the medium. It had the same amino terminal amino acid sequence as that of authentic CTB and was fully active in GM1 ganglioside binding assay. PMID- 8405931 TI - Chromosomal DNA from both flagellate and non-flagellate Bordetella species contains sequences homologous to the Salmonella H1 flagellin gene. AB - The genus Bordetella contains four species: two are non-motile, the human pathogens B. pertussis and B. parapertussis; and two are motile, the broad host range mammalian pathogen B. bronchiseptica, and the avian pathogen B. avium. The motility of the latter two species is due to peritrichous flagella. Here we show that strains of all four species contain DNA sequences homologous to flagellin genes. Two types of gene probe were hybridised to Bordetella chromosomal DNa in Southern blots: the structural gene for H1 flagellin of Salmonella typhimurium and an oligonucleotide derived from the conserved N-terminal amino acid sequences of various flagellin proteins. ClaI-digested DNa from all four Bordetella species hybridised with both probes in Southern blots, although each species gave a characteristic pattern of hybridisation. This indicates that the non-motile B. pertussis and B. parapertussis species contain non-expressed flagellin genes. PMID- 8405932 TI - Heterologous gene expression in Campylobacter coli: the use of bacterial luciferase in a promoter probe vector. AB - A novel promoter probe plasmid (pSP73) was constructed to allow the analysis of environmentally regulated gene expression in Campylobacter. The vector utilizes the luxAB genes from Xenorhabdus luminescens, which encode a thermostable luciferase, as reporters of gene expression. The utility of this reporter system was demonstrated by placing the expression of luxAB under the transcriptional control of the flaA gene promoter. PMID- 8405933 TI - An inexpensive infrared growth sensor array for detection of bacterial antibiotic susceptibility. AB - An inexpensive infrared sensor was constructed and used for the rapid testing of bacterial antibiotic susceptibility by detection of changes in absorbance at 950 nm. By comparing cultures of clinical isolates together with control strains (Escherichia coli NCTC 10418, Staphylococcus aureus NCTC 6571 or Pseudomonas aeruginosa NCTC 10662) after addition of an antibiotic, results on susceptibility were obtained within 3-5 h from the original plate culture. Representative strains of E. coli, P. aeruginosa, and S. aureus were tested successfully against ampicillin, penicillin, gentamicin or ciprofloxacin. PMID- 8405934 TI - Effects on toxicity of eliminating a cleavage site in a predicted interhelical loop in Bacillus thuringiensis CryIVB delta-endotoxin. AB - When activated by treatment with mosquito (Aedes aegypti) gut extract, the Bacillus thuringiensis CryIVB delta-endotoxin lysed A. aegypti cells in vitro. SDS-PAGE and N-terminal sequence determination showed that in addition to removal of the C-terminal half of the molecule, the activated toxin had undergone proteolytic cleavage at two internal regions producing 47-48-kDa and 16-18-kDa polypeptides. Aligning the CryIVB protein sequence with the crystallographic structure of the CryIIIA toxin suggested that one set of cleavages occurred in a region before the start of the N-terminal helical bundle and the second cleavage site occurred in a predicted loop between helices 5 and 6 in the bundle at arginine-203. To investigate the suggestion by Li et al. that interhelical proteolysis is important in the cytolytic mechanism of these toxins, arginine-203 was substituted by alanine. The mutated toxin now resisted proteolysis at this position and showed a marked decrease in cytolysis in vitro but an increase in larvicidal activity. PMID- 8405935 TI - The glutamine cyclotransferase reaction of Streptococcus bovis: a novel mechanism of deriving energy from non-oxidative and non-reductive deamination. AB - Streptococcus bovis deaminated glutamine by a mechanism that did not involve glutaminase. Since pyroglutamate and ammonia were the only end-products, it appeared that glutamine deamination was catalyzed by a cyclotransferase reaction. Stationary S. bovis cells had essentially no intracellular ATP or membrane potential (delta psi), however, when they were provided with glutamine, intracellular ATP and delta psi increased to 0.52 mM and 158 mV, respectively. When glutamine-energized cells were treated with N,N-dicyclohexylcarbodiimide (DCCD, 150 microM), there was an even greater increase in intracellular ATP (> 5 fold) and the delta psi was dissipated. Because toluene-treated cells produced ATP from ADP and Pi, it did not appear that the cell membrane was directly involved in glutamine-dependent ATP generation. The rate of ammonia production was directly proportional to the glutamine concentration, but the stoichiometry of ATP to ammonia was always 1 to 1. Based on these results, it appeared that glutamine was deaminated by glutamine cyclotransferase which was coupled to ATP formation. The membrane bound ATPase then used the ATP to create a delta psi. PMID- 8405936 TI - Isolation and characterization of Escherichia coli strains producing microcins of B and C types. AB - A screening of 11956 enterobacteria isolates resulted in selection of seven active microcin-producing strains. The microcins were shown to be peptides or their derivatives with a rather broad spectrum of activity, mainly against Gram negative bacteria. According to cross-immunity criteria, the microcins studied belonged to two of the previously suggested types, B (five strains) and C (two strains). Those of type B could be further classified into two subtypes on the account of differences in the spectrum of antibacterial activity. In five cases out of seven the microcin-producing ability has been attributed to plasmids that the strains harboured. The effect of microcins on sensitive cells was shown to depend on ompR and ompF gene products. PMID- 8405937 TI - Expression of an iron-regulated hemolysin by Edwardsiella tarda. AB - The ability of Edwardsiella tarda to hemolyse red blood cells was investigated. Most E. tarda strains (> 80%) produced a hemolysin when assayed by either an agar overlay or contact-dependent hemolysis technique. This activity was cell associated (CAH) and not released into the culture supernatant under routine conditions. When quantified, E. tarda strains significantly produced 30-40-fold higher levels of hemolytic activity against guinea pig, sheep, or rabbit erythrocytes than either E. hoshinae or E. ictaluri. When grown under iron restricted-conditions in the presence of ethylenediamine di(o-hydroxyphenylacetic acid), hemoglobin, hematin and hemin were found to stimulate growth in both liquid and agar bioassays. Hemolysin activity could be released from selected E. tarda strains when grown in L broth supplemented with EDDA; hemolytic activity was 3- to > 40-fold under these conditions when compared to L broth alone. Preliminary characterization of the hemolysin of strain ET-13 indicates that it is a heat-labile protein with active sulphydryl and thiol groups. These results indicate that, in addition to its invasive capabilities, E. tarda produces a hemolysin which is at least partially regulated by the relative availability of iron and may play a role in human disease. PMID- 8405938 TI - Membrane location of the Ti plasmid VirB proteins involved in the biosynthesis of a pilin-like conjugative structure on Agrobacterium tumefaciens. AB - The virB operon of the Agrobacterium tumefaciens Ti plasmid encodes 11 proteins. Specific antisera to VirB2, VirB3 and VirB9 were used to locate these virulence proteins in the A. tumefaciens cell. Immunoblot analysis located VirB2 protein to the inner and outer membranes; VirB3 and VirB9 were likewise associated with both membranes, but mainly in the outer membrane. VirB2 is processed from a 12.3-kDa protein into a 7.2-kDa polypeptide. Such sized protein results from cleavage at residue Ala47, upstream of which two additional alanine residues Ala45-Ala46 are contained and bearing resemblance to a signal peptide peptidase-I cleavage sequence. VirB2 and VirB3 sequences are strikingly similar to the pilin biosynthetic proteins TraA and TraL encoded by the tra operon of F and R1-19 plasmids. Since traA encodes a propilin that is cleaved into a 7.2-kDa conjugative pilin product and since this cleavage site is present in both TraA and VirB2, we propose that virB2 encodes a pilin-like protein which together with VirB3 and VirB9 as well as other VirB proteins may be used for interkingdom T-DNA transfer between bacteria and plants. PMID- 8405939 TI - Investigation of the Pseudomonas aeruginosa ampR gene and its role at the chromosomal ampC beta-lactamase promoter. AB - The complete sequence of the Pseudomonas aeruginosa beta-lactamase regulator gene ampR has been determined. The amino acid sequence encoded by this gene is compared with other members of the LysR family of transcription factors. The P. aeruginosa ampC promoter was subcloned and gel mobility shift assays were used to demonstrate specific binding of AmpR to the ampC promoter region. PMID- 8405940 TI - Studies on the pathophysiological mechanism of Campylobacter jejuni-induced fluid secretion in rat ileum. AB - Calcium has been reported to play an important role in regulating the intestinal electrolyte transport via Ca2+/calmodulin (CaM) and/or protein kinase C (PKC) systems. The role of Ca2+, CaM and PKC in the pathogenesis of Campylobacter jejuni-induced fluid accumulation was studied in vivo in ligated rat ileal loops. Calcium ionophore A23187 (5 microM) and PKC activator, phorbol-12-myristate-13 acetate (PMA, 100 micrograms kg-1) when injected alone induced fluid accumulation in the control loops. However, these modulators did not enhance further C. jejuni induced fluid accumulation when injected along with C. jejuni live culture in the experimental loops. Both 1-verapamil (100 microM) and PKC antagonist, H-7 (15 micrograms/ml-1) significantly reduced C. jejuni-induced fluid accumulation (P < 0.001). The effect of CaM antagonist W-7 (60 microM) on C. jejuni-induced fluid secretion was not significant (P > 0.05). Our findings suggest that both Ca2+ and PKC appear to be the important second messengers involved in the stimulation of intestinal fluid accumulation in C. jejuni infection. PMID- 8405941 TI - Analysis of a cloned Francisella tularensis outer membrane protein gene and expression in attenuated Salmonella typhimurium. AB - We have determined the nucleotide sequence of fopA from Francisella tularensis. Using the polymerase chain reaction fopA was detected in high and low virulence biotypes of F. tularensis. fopA was stably maintained in pBluescript in attenuated Salmonella typhimurium where FopA was expressed and located in the outer membrane. This recombinant will be suitable for studies on the role of FopA in immunity against tularaemia. PMID- 8405942 TI - Inhibition of mitogen-induced proliferation of spleen lymphocytes is correlated with the induction of cell-mediated immunity in Salmonella infection in mice. AB - The proliferation of murine spleen cells stimulated by a T-cell mitogen such as phytohemagglutinin (PHA) or concanavalin A (ConA) was significantly suppressed when the mice were immunized with either the viable cells or the sonicate of Salmonella typhimurium but not of Escherichia coli. The suppression of T-cell proliferation caused by the sonicate of S. typhimurium was completely restored by addition of phorbol 12-myristate-13-acetate (PMA), an activator of protein kinase C (PKC). Western blots using anti-phosphotyrosine antibodies showed that the mitogen-induced tyrosine phosphorylation of 120-, 106-, 94-, 76-, 68- and 57-kDa proteins in murine splenic T-cells was inhibited in the mice immunized with the viable cells but not the sonicate of S. typhimurium. These results suggest that the inhibition caused by the sonicate involves suppression of PKC activity, whilst that produced by viable cells involves down-regulation of tyrosine phosphorylation, and both inhibitions correlate with the induction of cell mediated immunity in mice, as evidenced by the induction of delayed-type hypersensitivity reactions. PMID- 8405943 TI - Expression of Aspergillus oryzae alpha-amylase gene in Saccharomyces cerevisiae. AB - A fragment containing the full length cDNA from Aspergillus oryzae alpha-amylase has been amplified by PCR using specific synthetic oligonucleotides. The amplified cDNA was designed to favour its expression in yeast by modifying its upstream untranslated region. It was subcloned in the expression vector pYEX alpha 1, placed under the control of the yeast CYC1-GAL10 promoter and used to transform Saccharomyces cerevisiae. Cells were then able to express and secrete active alpha-amylase to the medium in a regulated fashion. The recombinant enzyme had similar electrophoretic mobility and catalytic properties to the original A. oryzae alpha-amylase. PMID- 8405944 TI - Investigation of the role of the cydD gene product in production of a functional cytochrome d oxidase in Escherichia coli. AB - The cydD gene is needed for the formation of a functional cytochrome d oxidase in the aerobic respiratory chain of Escherichia coli. In this paper we demonstrate that transcription from a cydA-lacZ gene fusion is not significantly affected in a cydD mutant. This, together with the finding that subunit I of cytochrome d is present in cytoplasmic membranes of a cydD mutant, rules out a role for CydD in the regulation of cytochrome d expression or the assembly of its polypeptides into the membrane. The activity of the haem d-containing catalase HP II was found to be similar in a cydD mutant and the wild-type. Therefore, if CydD has a role in haem d biosynthesis it must be in a unique step only associated with the synthesis of the haem d prosthetic group of cytochrome d. PMID- 8405945 TI - Genetic evidence of a new flocculation suppressor gene in Saccharomyces cerevisiae. AB - The flocculation character in strain IM1-8b of Saccharomyces cerevisiae is controlled by a single and dominant gene shown to be allelic to FLO1. Such a gene has been both mitotically and meiotically mapped on the right arm of chromosome I at 4.7 cM from PHO11. The phenotype was suppressed by a single gene of wide distribution among non-flocculent strains (proposed as fsu3) that, however, was unable to suppress other FLO1 genes in other flocculent strains. PMID- 8405946 TI - Presence of carnitine acetyltransferase in peroxisomes and in mitochondria of oleic acid-grown Saccharomyces cerevisiae. AB - Activity of carnitine acetyltransferase was detected in glucose- and oleic acid grown Saccharomyces cerevisiae. Oleic acid-grown cells showed a ten-fold higher activity than glucose-grown cells. Subcellular fractionation of oleic acid-grown cells showed that carnitine acetyltransferase was present in peroxisomes, mitochondria, and cytosol. The results suggested the plausible presence of an 'acetylcarnitine shuttle' in this yeast, as in the case of an n-alkane assimilating yeast, Candida tropicalis. PMID- 8405947 TI - Purification and molecular properties of an alpha-galactosidase synthesized and secreted by Aspergillus nidulans. AB - alpha-Galactosidases from mycelial extract and culture filtrate of Aspergillus nidulans have been purified to homogeneity and utilised to obtain polyclonal antibodies anti-alpha-galactosidase. The enzymatic characteristics and the cross reactivity of the antibodies suggest that alpha-galactosidases isolated from the two sources were the same enzyme. Thus, A. nidulans synthesized and secreted only one enzymatic form of alpha-galactosidase which is a multimeric enzyme of 370 kDa composed of four monomers of 87 kDa and a pI of 6.3. The optimum temperature of activity was 50 degrees C and the optimum pH 4-5. The enzyme was stable over a wide range of pH but quite unstable to temperature. alpha-Galactosidase of A. nidulans is a very specific enzyme, it is active only on p-nitrophenyl-alpha-D galactoside (PNPG), melibiose and raffinose. When PNPG was utilised as substrate melibiose, raffinose, galactose and glucose were competitive inhibitors of the activity. PMID- 8405948 TI - Purification and characterization of EpiA, the peptide substrate for post translational modifications involved in epidermin biosynthesis. AB - For the investigation of enzymes involved in epidermin biosynthesis it is necessary to produce sufficient amounts of preepidermin (EpiA) as a substrate and to design EpiA detection systems. Therefore, EpiA was expressed in Escherichia coli using a malE-epiA fusion. The identity of purified EpiA was confirmed by ion spray mass spectrometry and amino acid sequencing. For EpiA detection, anti-EpiA antisera were raised. Upon prolonged incubation, factor Xa not only cleaved EpiA from the fusion protein, but also less efficiently cleaved EpiA internally between R-1 and I+1. The internal factor Xa cleavage site of EpiA was masked by altering the sequence -A(-4)-E-P-R(-1)- to -A(-4)-E-P-Q(-1)- by site-directed mutagenesis. PMID- 8405949 TI - Rapid isolation of genes from bacterial lambda libraries by direct polymerase chain reaction screening. AB - A method for the direct screening of bacterial lambda libraries by polymerase chain reaction technology has been developed. This technique permits the identification and isolation of specific DNA sequences without the need for any filter hybridisation or radioactive probing. This strategy has been used to isolate a gene encoding lactate dehydrogenase from a Lactococcus lactis lambda library. PMID- 8405950 TI - Purification and characterization of membrane-bound CO-reactive hemoprotein from Tetrahymena pyriformis mitochondria. AB - A CO-reactive hemoprotein was purified from the mitochondrial membrane fraction of Tetrahymena pyriformis. It showed absorption peaks at 615 and 455 nm in the reduced form and an alpha peak at 565 nm in the pyridine ferrohemochrome spectrum. Although the spectral properties were apparently similar to those of 'cytochrome a620' which was previously proposed as a mitochondrial terminal oxidase in T. pyriformis, it did not contain any molecules of heme a or copper atoms. Further, it showed neither cytochrome c oxidase nor cytochrome c peroxidase activity. These results suggest that 'cytochrome a620' may not be the terminal oxidase in the mitochondrial respiratory chain of T. pyriformis. PMID- 8405951 TI - Detection of Norwalk virus in the UK by the polymerase chain reaction. AB - We have developed a polymerase chain reaction for the detection of Norwalk virus using the published sequence of the virus RNA dependent RNA polymerase gene and have used this to clone and sequence this region of a virus from a UK outbreak. We have applied this method to a panel of UK Norwalk-like viruses using both Tet z and Taq DNA polymerases and found that amplification produces a multiplicity of bands from stool samples. However, in combination with Southern blotting, Taq polymerase amplification detected virus in 13 of a panel of 30 clinical samples known to contain these viruses and also detected astroviruses in a mixed infection. Amplification using Tet-z DNA polymerase was less efficient (6/30) and detected predominantly viruses typed as UK type 2 by solid phase immune electron microscopy. PMID- 8405952 TI - Influence of pulsing and postpulsing media tonicity on electrotransformation of intact yeast cells. AB - The maximal transformation yield of intact yeast cells in hypotonic medium was obtained by application of nine pulses with a duration of 990 microseconds at 2.5 kV cm-1. Pulsation at the same electrical parameters in isotonic solution did not lead to any transformation and the electropermeability decreased by 50%. The transfer of cells, 1 min after pulsation in hypotonic medium, into media with different tonicity led to an increase of the number of transformed cells, depending on the sorbitol concentration of up to 250 mM. Further augmentation of the tonicity of postpulse medium in the range 330-1000 mM provoked strong decrease of transformation. This effect was present even when cells were resuspended in isotonic medium 30 min after pulsation. PMID- 8405953 TI - Construction and overexpression in Escherichia coli of genetically engineered derivatives of penicillin-binding protein 2' of Staphylococcus epidermidis. AB - Removal of the putative amino-terminal membrane spanning region of penicillin binding protein 2' (PBP-2') of Staphylococcus epidermidis WT55 was carried out by truncating the amino terminus-coding end of the mecA gene. PCR and site directed mutagenesis were used to introduce unique restriction sites at position 68 (HindIII) and at position 80 (NcoI) of the mecA gene, respectively. The coupling of the shortened coding regions to the trc promoter and gene fusion to the lacZ gene, aimed to facilitate subsequent protein purifications, resulted in strong expression in the cytoplasm of Escherichia coli and partial sequestration into insoluble protein granules. The truncated PBP-2' retained its penicillin-binding ability and also bound the monoclonal antibody directed against PBP-2' of Staphylococcus aureus. PMID- 8405954 TI - Bacillus subtilis mutant deficient in the major autolytic amidase and glucosaminidase is impaired in motility. AB - The purified autolytic endo-beta-N-acetylglucosaminidase of Bacillus subtilis AC327 was cleaved with cyanogen bromide, and the N-terminal amino acid sequence of one of the peptide fragments was determined. Then, a DNA fragment containing a part of the glucosaminidase gene was cloned into Escherichia coli JM109 using synthetic oligonucleotides as probes whose sequences had been deduced from the N terminal amino acid sequence. Zymographic analysis showed that the resultant glucosaminidase-deficient strain lacked a 35-kDa lytic band in addition to a 90 kDa lytic one corresponding to the glucosaminidase. A double mutant strain deficient in the major two autolysins (amidase and glucosaminidase) exhibited greatly impaired motility on a swarm plate whereas the single mutant strains were motile. PMID- 8405955 TI - Brucella group 3 outer membrane proteins contain a heat-modifiable protein. AB - Brucella melitensis and B. ovis outer membrane blebs contained a protein displaying a temperature-dependent molecular mass upshift from 25 kDa to 30 kDa. A fraction of the protein tightly bound to LPS did not show the molecular mass upshift which was also blocked by exposure of the protein to Zwittergent 314. The B. melitensis heat-modifiable protein and Escherichia coli OmpA shared antigenic determinants. These data indicate that the Brucella group 3 outer membrane proteins belonged to the OmpA family of proteins. PMID- 8405956 TI - Induction of autolysis in starved Escherichia coli cells by seminalplasmin, an antimicrobial protein from bovine seminal plasma. AB - A pair of relA+ and relA E. coli strains, otherwise isogenic, were studied with regard to the susceptibility of starved cells to lysis induced by the natural peptide seminalplasmin. Starved relA cells were more sensitive to seminalplasmin induced lysis when compared to starved relA+ cells. Nevertheless, pronounced lysis of starved relA+ cells was observed with increase in the concentration of seminalplasmin. In contrast, ampicillin could not lyse starved relA+ cells even at very high concentrations. Further, seminalplasmin could cause loss of viability and degradation of peptidoglycan in starved relA+ cells. These observations suggest that, unlike many other antibiotics, seminalplasmin can induce autolysis under the conditions of a stringent response. PMID- 8405957 TI - Transformation of 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8- dihydroxyquinoline by quinoline-grown resting cells of Pseudomonas putida 86. AB - Resting cells of Pseudomonas putida strain 86 were grown on quinoline transformed 2-chloroquinoline to 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline which was not converted further. 7,8-Dioxygenating activity was present when the enzymes of quinoline catabolism were induced. Quinoline-grown cells of strain 86 treated simultaneously with 2-chloroquinoline and D-(-)-threo-chloramphenicol to prevent protein biosynthesis also formed the cis-7,8-dihydrodiol of 2-chloroquinoline. Succinate-grown resting cells did not oxidize 2-chloroquinoline. Acid-catalyzed decomposition of 2-chloro-cis-7,8-dihydro-7,8-dihydroxyquinoline predominantly yielded 2-chloro-8-hydroxyquinoline. By analogy, accumulation of the putative dead-end metabolite 1H-8-hydroxy-2-oxoquinoline during growth of P. putida 86 on quinoline is suggested to likewise result from dehydration of the 7,8-dihydrodiol of 1H-2-oxoquinoline. PMID- 8405958 TI - Characterization of the transferrin-iron uptake system in Neisseria meningitidis. AB - The transferrin-iron uptake system of six Neisseria meningitidis strains was characterized using 125I-transferrin in receptor assays and 55Fe-loaded transferrin in uptake assays. Receptors for transferrin varied among the strains both in number (from 700 to 4700 receptors per cell) and in their affinity constants for the protein (Ka ranged from 0.7 x 10(7) to 4.0 x 10(7) l mol-1). Neither receptor numbers nor affinity constants were significantly different in carrier and invasive strains, although the Ka seem to be somewhat higher in the latter. Iron uptake from transferrin was also variable among the strains, but showed the same lack of correlation with their origin. PMID- 8405959 TI - The antibacterial activity of haemin compared with cobalt, zinc and magnesium protoporphyrin and its effect on potassium loss and ultrastructure of Staphylococcus aureus. AB - The unique antibacterial properties of Fe-protoporphyrin (haemin) on Staphylococcus aureus, compared to Co-protoporphyrin (Co-PP), Mg-protoporphyrin (Mg-PP) and Zn-protoporphyrin (Zn-PP) are described. Only haemin (20 microM) exhibits a strong light-independent antibacterial effect on S. aureus; the other metalloporphyrins, Co-PP, Mg-PP or Zn-PP, have no antibacterial effect in the dark. Only light photosensitization of Mg-PP-treated cells resulted in the inhibition of the bacterial growth, while Co-PP or Zn-PP were photodynamically inactive. A notable effect of haemin on inactivation of S. aureus was the induction of immediate ion fluxes as determined by X-ray microanalysis (XRMA) of fast-frozen cells. A marked efflux of K (96%) and Cl (94%) was expressed immediately as determined by X-ray microanalysis of S. aureus cells treated with haemin for 5 min. Only 48% loss of Na was detected in the cells under these treatment conditions, while P content was increased by 150%. Electron microscopy analysis revealed the appearance of a mesosome-like structure connected to the new septa, filamentous chromosome and arrays of aggregated ribosomes in the cytoplasm. We propose that haemin has multiple cellular targets for its oxidative effect in S. aureus. PMID- 8405960 TI - The bifunctional NadR regulator of Salmonella typhimurium: location of regions involved with DNA binding, nucleotide transport and intramolecular communication. AB - NadR is the repressor protein that controls the expression of genes for NAD synthesis. It is also believed to be involved in nucleotide transport. Point mutations conferring different phenotypes were localized to six different regions within the nadR gene. That mutations affecting repression and transport all mapped within nadR confirms the bifunctional model for NadR action. The clustering of these mutations and 2 fusions revealed that those affecting repression lie in the amino terminal while those affecting transport occur in the carboxy-terminal. Mutations resulting in superrepression occurred within a central region of NadR that probably senses NAD concentrations. This region is predicted to direct the transition between NadR transport and repressor conformations. PMID- 8405961 TI - Molecular cloning and nucleotide sequence analysis of the maltose-inducible porin gene of Aeromonas salmonicida. AB - The gene for the Aeromonas salmonicida maltose-inducible porin (maltoporin) was cloned into phagemid pTZ18R in two restriction fragments, 0.6-kb PstI/KpnI and 1.7-kb SphI, of genomic DNA and their nucleotide sequences were determined. Open reading frames of 1329 and 1335 bp translated into sequences of 443 and 445 amino acids, with a 23 or 25 amino acid signal sequence and a 420 amino acid mature protein of molecular mass 46424 Da. Putative ribosome binding sites, AGGA and GGGAA, occurred 9 bp upstream of two possible ATG initiation codons. The A. salmonicida gene product showed a high degree of similarity with Escherichia coli LamB, and codon usage was very similar to that of another A. salmonicida outer membrane protein but markedly different from those of extracellular proteins. PMID- 8405962 TI - Outer membrane complex proteins of Chlamydia pneumoniae. AB - The protein composition of the outer membrane complex (OMC) of Chlamydia pneumoniae strain AR-39 was analyzed by metabolic labeling with [35S]methionine and [35S]cysteine. Cysteine-rich proteins with molecular masses of 98, 60 doublet, 39.5 (MOMP) and 15.5 kDa were found in the OMC of C. pneumoniae. The cysteine-rich proteins of the OMCs of the three Chlamydia species showed specific reaction patterns by immunoassay and autoradiography to rabbit or turkey immune sera. Recognition of the MOMP and 60-kDa proteins of the three species was cross reactive. However, the C. pneumoniae 98-kDa protein was recognized by anti-C. pneumoniae (AR-39) and anti-C. psittaci (TT3) immune sera. None of the immune sera recognized the 12-kDa cysteine-rich complex. PMID- 8405963 TI - Molecular cloning and sequencing of infC, the gene encoding translation initiation factor IF3, from four enterobacterial species. AB - Translation initiation factor IF3 plays a crucial role in initiation of protein synthesis in bacteria. In order to elucidate the IF3 structural elements required for these functions, the evolutionary conservation of IF3 and its gene, infC, was investigated. Homologous infC sequences from Salmonella typhimurium, Klebsiella pneumoniae, Serratia marcescens and Proteus vulgaris were amplified by the polymerase chain reaction and sequenced. Analysis of these sequences, as well as that from Bacillus stearothermophilus, revealed several regions (e.g. residues 62 73 and 173-177) of absolute sequence conservation, suggesting an important role for these regions in IF3 function. PMID- 8405964 TI - Molecular subtyping within a single Salmonella typhimurium phage type, DT204c, with a PCR-generated probe for IS200. AB - We report primers and conditions for the generation by PCR of a probe for the DNA insertion element IS200. This probe was shown to be suitable for genotypic subtyping within a single phage type of Salmonella typhimurium. A collection of isolates of DT204c, a phage type implicated in the spread of multiple drug resistance in bovine animals and to man, was analyzed. Three IS200 profiles, corresponding to related chromosomal genotypes were characterized in DT204c. Molecular discrimination within a single phage type of S. typhimurium has general significance for genotypic typing, and for the definition of epidemiological clonality in Salmonella. PMID- 8405965 TI - Transcript mapping of the rubredoxin gene from Clostridium pasteurianum. AB - The transcripts of the rubredoxin gene from C. pasteurianum have been shown to have a size of approx. 230 bases by Northern blotting techniques. The transcription start has been located within 41 bases upstream of the initiator codon. The data demonstrate that the rubredoxin gene is monocistronic. Two of the three open reading frames occurring upstream of the rubredoxin gene are adjacent and appear to encode a thioredoxin (or glutaredoxin) reductase and a thioredoxin (or glutaredoxin). PMID- 8405966 TI - The Alcaligenes eutrophus ldh structural gene encodes a novel type of lactate dehydrogenase. AB - The lactate dehydrogenase gene, ldh, of Alcaligenes eutrophus H16 was identified on a 14-kbp EcoRI restriction fragment of a genomic library in the cosmid pHC79 by hybridization with a 50-mer synthetic oligonucleotide which was derived from the N-terminal amino acid sequence of the purified enzyme. Recombinant strains of Escherichia coli JM83, which harboured a 2.0-kbp PstI subfragment in pUC9-1, expressed LDH at a high level, if ldh was downstream from and colinear to the E. coli lac promoter. The nucleotide sequence of a region of 4245 bp revealed several open reading frames which might represent coding regions. One represented the ldh gene. The amino acid sequence deduced from ldh exhibited 29% and 36% identity to the L-malate dehydrogenase of Methanothermus fervidus and to the putative translation product of an E. coli sequence of unknown function, respectively. The ldh was separated by short intergenic regions from two other open reading frames: ORF5 was located downstream of and colinear to ldh, and its putative translational product revealed 38 to 56% amino acid identity to penicillin-binding proteins. ORF3 was located upstream of and colinear to ldh, and its putative gene translational product represented a hydrophobic protein. A sequence, which resembled the A. eutrophus alcohol dehydrogenase promoter, was detected upstream of ORF3, which most probably represents the first transcribed gene of an operon consisting of ORF3, ldh and ORF5. PMID- 8405967 TI - Relationships with parents and friends: cross-sectional and retrospective accounts. AB - We investigated Turkish high school and university students' perceptions of their relationships with their parents and friends in two studies using cross-sectional and retrospective methods. Subjects in both studies rated 31 items describing their relationships with one significant other from two perspectives: their own point of view and that of their relationship partner (mother, father, or best same-sex friend). Results indicated age-related trends for perceptions of development of relationships with significant others. Retrospective reports revealed greater degrees of perceived equity of self-favoring inequity than did cross-sectional reports. PMID- 8405968 TI - Adaptive versus maladaptive emotional tension. AB - The problem of emotional tension is usually discussed only with regard to its negative (maladaptive) aspects. The positive (adaptive) function of emotional tension, however, is as important as its negative function. In this article, we have examined psychophysiological outcomes of adaptive versus maladaptive emotional tension with respect to opposite forms of behavior (search activity and renunciation of search) that have an opposite outcome on body resistance and performance. Some traditional psychophysiological problems, for example, the law of initial values (Wilder, 1931), the difference between orienting and defensive reaction, or between anxiety and panic behavior, are revised on the basis of the assumption of adaptive and maladaptive emotional tension. PMID- 8405969 TI - Contagion: a theoretical and empirical review and reconceptualization. AB - We have reviewed theories and research in the area of contagion with an emphasis on definitions of contagion. The review shows that a great deal of the confusion surrounding the term is due to the fact that the phenomena involved in contagion are extremely heterogeneous, yet they typically have been placed under homogeneous rubrics. Accordingly, we propose herein that contagion should be conceptualized as a general type, social contagion, and three subtypes: disinhibitory, echo, and hysterical. In this article, we have distinguished social contagion and its subtypes from other types of social influence phenomena and reclassified theoretical and research articles under the newly proposed definitions. The proposed reconceptualization shows how contradictions in the literature can be resolved by distinguishing the type of contagion in question and provides the foundation for a more comprehensive and useful psychology of contagion. PMID- 8405970 TI - [Theoretical analysis of some features of compensatory substitution in stem regions of transport RNA]. AB - Some families of isoacceptor tRNAs were studies using VOSTORG software package for phylogenetic analysis. While analysing the evolutionary trees in was shown that the fixation of double substitutions AU<-->GC in spiral double-stranded regions passes two stages with intermediate station, such as non-canonical base pairs. In all the families studied double substitutions can be explained not only by passages through slightly deleterious GU pairs. Comparative analysis of B2 like repeats shown the absence of compensatory substitutions in the parts which are homologous to RNA stem regions. The data obtained let us assume that compensatory substitutions in the RNA helical regions represent an expressive example of non-directed adaptive molecular coevolution. PMID- 8405971 TI - [Nucleotide sequence and properties of an inverted repeating element of a Bordetella pertussis chromosome]. AB - The repeated sequence from Bordetella pertussis chromosome was cloned using the method described by Ohtsubo. The sequences of the characterized B. pertussis chromosome are homologous to the previously described sequences and analogous in the structure to the already known IS elements. The parental recombinant plasmids containing the RS were unstable and segregated to the plasmids of different structure. The segregants' structure is characterized in this paper. It is shown in our study that the RS element is able to stimulate intragenomic rearrangements, such as deletions. It is shown that at least one deletion begins precisely after 3' end of RS and terminates with the sequence which is completely homologous to ten terminal nucleotides of this one. Probably, RSs stimulate at high frequency the formation of deletions, which appear to be the result of recA independent site-specific recombination between short direct repeats. PMID- 8405972 TI - [The effect of certain Escherichia coli genes on the appearance of the TCS phenotype, conferred by plasmid RP4 with an integrated genome of the D3112 Pseudomonas aeruginosa phage]. AB - The possibility of the SOS system activation caused by introduction of a hybrid plasmid RP4::D3112 (where D3112 is a genome of the transposable phage of Pseudomonas aeruginosa) into Escherichia coli was examined. It has been shown previously that the presence of this plasmid confers to E. coli a so called TCS phenotype: E. coli (RP4::D3112) forms normal colonies and grows at 42 degrees C but does not divide and becomes filamentous at 30 degrees C, probably because of E. coli DNA damages generated in the course of D3112 replication-transposition. It was shown that the level of prophage lambda induction is not elevated during the growth of E. coli (lambda) (RP4::D3112) cells at 30 degrees C. The character of the TCS phenotype was not affected by lex A3 or lex A51 mutations (which cause correspondingly non-inducibility or constitutive expression of the SOS regulon). It was concluded therefore that the TCS phenotype is not related to induction of the SOS response. It was found also that the mutation to heat shock gene dnaJ (dnaJ259) is a cause for significant decrease in phage D3112 production in E. coli dnaJ259 (RP4::D3112) cells. The DNA hybridization data of labelled RP4 and D3112 DNA with total E. coli dnaJ259 (RP4::D3112) DNA suggest that the dnaJ259 probably affected the late D3112 functions. PMID- 8405973 TI - [The effect of inhibitors on induced mutagenesis]. AB - The mutagenic effect induced by mitomycin C (MC) 20 mg/ml in early, middle and late G1-phase of Crepis capillaris seeds was modified by DNA synthesis inhibitors -caffeine 5 mM, hydroxyurea (HU) 5 x 10(-3) M, 1-beta-D-arabinofuranosylcytosine (araC) 1 x 10(-3) M and aphidicolin (APC) 0.5 mg/ml in the DNA post-synthesis phase (G2). It was shown that under the action of MC in early G1 and the inhibitors in the G2 the total number of aberrations for fixations is lower in all the cases than under the action of MC only. The action of MC in the middle phase results in the differential effect of the inhibitors. HU and APC were shown to potentiate the effect induced by MC and ara-C to reduce it. In both cases, caffeine at the dose 5 mM had small effect on DNA damages induced by MC in early and middle G1. Under the action of caffeine and HU in the G2-phase the crosslinks induced by MC in the late G1 result in aberrations of chromosomal type, illegal for the S-dependent mutagenesis. On the whole, the process of mutation formation turned out to be sensibilized. The specificity of mutagenesis under the action of MC suggests the appearance of mutations of the chromosomal type after the DNA synthesis in G2 phase. PMID- 8405974 TI - [Analysis of chimeric mice bearing the fused mutation]. AB - Interaction of cells with different genotypes and expression of Fused (Fu) gene in chimaeric mice were studied. Genetic analysis of 20 chimaeras showed that gonads of chimaeras in our experiments consisted of white component cells. The analysis of Fu gene expression in chimaeric mice supported the conclusions of the previous papers that mutations of Fu gene appeared not in the result of the hypofunction of normal allele. Two hypotheses were proposed: 1) the product of Fu gene migrates out of the mutant cells and changes the phenotype of the cells with normal allele; 2) suppressors from C57BL/6 mice influence on the penetrance of Fu gene only on the intracellular level. PMID- 8405975 TI - [Radiation-induced mitotic crossing-over in the pronucleus of Drosophila males]. AB - Exchanges in nondisjoined homologous chromosomes of irradiated mature spermatozoa were obtained. Exchanges absent in the control experiment where one of those chromosomes was irradiated in mature sperm and the other (non-irradiated) comes from female. Those results coupled with Abeleva et. al. data show that the exchanges observed were formed in male pronucleus. PMID- 8405976 TI - [Genetic analysis of the interaction of mutations in two genes, controlling expression of MDG4]. AB - Transcription of the retrovirus-like mobile element mdg4 (gypsy) in controlled by the protein product of the suppressor of Hairy wing which interacts with the mdg4 enhancer, and by the protein product of the mdg4 modifier which seems to interact with the su(Hw) protein. In the present work, we studied interaction of mod(mdg4) and su(Hw) mutations in phenotypic expression of mutations induced by mdg4 insertion. Different types of inhibitory effect of mdg4 enhancer on transcription of the target gene were detected. Most frequently, the inhibitory depends on the presence of the mod(mdg4) protein. On of the two acidic domains of su(Hw) protein may be removed without changing the effect. Additional removal of the leucine zipper domain strongly reduces the effect. The second type of inhibition depends of or decrease in the concentration of the mod(mdg4) protein. The su(Hw) protein should have both acidic domains in this case. Sometimes, the su(Hw) protein lacking the C-end acidic domain may activate transcription of a target gene, and this effect does not depend on the presence or absence of the mod(mdg4) protein. The effect may be increased by removal of leucine zipper. Binding of the mod(mdg4) protein to the su(Hw) protein does not depend of the C-end acidic domain and leucine zipper. PMID- 8405977 TI - [Population-genetic characteristic of the rural population of southern Tajikistan]. AB - The results of genetic and demographic study of the inhabitants in south Tadjikistan are presented. It had been shown that the population studied is a typical rural population of Middle Asia and is characterized by the extended reproductive capacity, homogeneity of national composition and a high proportion of consanguineous marriages. Genetic variation in blood groups (AB0, MN, Rh-D, P1), serum proteins and red cell enzyme systems (Tf, Gc, Hp, PGM1, PGD, EsD, AcP1) was examined. A comparative analysis of gene frequencies in the population studied and some other Middle Asia populations has been carried out. PMID- 8405978 TI - [Polymorphism of mitochondrial DNA in the Russian population of Russia]. AB - Restriction enzyme fragment patterns in the D-loop and deletion-insertion polymorphism in the V non-coding region of human mitochondrial DNA (mtDNA) were analysed in a Russian population using the polymerase chain reaction. Polymorphisms were detected and mtDNA classified into 26 types using eight enzymes--AvaII, BamHI, EcoRV, KpnI, HaeIII, RsaI, Cfr13I, Sau3AI and Asian specific deletion and insertion. The allele and mitotype frequencies were determined. The data obtained for Russian population and the literature data are comparatively studied. PMID- 8405979 TI - [Formalizing the correlative selection of pedigrees]. AB - A method for ascertainment formalization is proposed when the trait subjected to a segregation analysis is a correlate to the trait on the basis of which the ascertainment of pedigrees is carried out. The method is a modified AAF method proposed by Ewens and Shute. PMID- 8405980 TI - [Planning selection for informativity of linkage analysis of separate pedigrees]. AB - Comparison of Kullback's information on nuclear pedigrees with different numbers of offsprings was performed in a case when a marker gene was codominant and the recessive or dominant control with complete penetrance was supposed for the localized gene. This comparison resulted in some conclusions about optimization of the sample collection. These conclusions are found to be invariant with respect to the recombination fraction value determined by the simple alternative hypothesis, to the population frequencies of the genes considered and, within certain limits, to the disequilibrium parameter values. PMID- 8405981 TI - [Dependence of the load of hereditary diseases on the level of endogamy of the Russian population]. AB - Relationship between the index of endogamy and a random component of inbreeding in Russian populations was studied. It was shown that significant correlation (r = 0.74 +/- 0.24) exists between these parameters. Correlation coefficient between the index of endogamy and the load of autosome recessive pathology makes up 0.98 + 0.11. It is suggested that the index of endogamy could be used as a quantitative measure of genetic structure of the population where it is not possible to receive the inbreeding value using the routine methods. PMID- 8405982 TI - 1992 Genetics Society of Canada Award of Excellence Lecture: Genes, science, and society. PMID- 8405983 TI - The rbcL gene sequence from chestnut indicates a slow rate of evolution in the Fagaceae. AB - The nucleotide sequence was obtained for the chloroplast gene coding for the large subunit of the ribulose 1,5-bisphosphate carboxylase (rbcL) of chestnut (Castanea sativa Mill.), a member of the woody family Fagaceae. Amplification primers downstream and upstream the rbcL open reading frame are also described. By comparing with other angiosperm sequences, we show that the rate of evolution of rbcL in the family Fagaceae is much slower than that observed for the families of annuals analyzed. PMID- 8405984 TI - A developmental and molecular analysis of Cdc2 mutations in Drosophila melanogaster. AB - In fission yeast, the product of the cdc2 gene is required both for entry into S phase and mitosis. Homologs of cdc2 have been isolated from a number of metazoans, but in general they have not been amenable to genetic analysis. Here we describe P element transposon tagging of Cdc2 in Drosophila melanogaster and the analysis of 10 Cdc2 mutants. The recessive lethality of Cdc2P is associated with a P element located in the 5' untranslated region of the gene. Seven other alleles have unique single base pair substitutions in the coding region of Cdc2. One allele, Cdc2B47, is mutated in the splice donor site of exon 1. Most mutations in Cdc2, including the presumptive null allele Cdc2B47, die at the pupal stage, suggesting that the maternally supplied Cdc2 gene product drives earlier cell divisions. The phenotypes of our mutants are consistent with a role for Cdc2 in cell proliferation; however, we did not observe any perturbation of the endoreduplication cycle associated with the acquisition of polyteny. PMID- 8405985 TI - Genetic diversity and phylogeny analysis of Azolla based on DNA amplification by arbitrary primers. AB - The polymerase chain reaction was used to amplify random sequences of DNA from 25 accessions of Azolla to evaluate the usefulness of this technique for identification and phylogenetic analysis of this aquatic fern. Accessions were selected to represent all known species within the genus Azolla and to encompass the world-wide distribution of the fern. Primers of 10 nucleotides with 70% G + C content were used to generate randomly amplified polymorphic DNA from the symbiotic Azolla-Anabaena complex. Twenty-two primers were used and each primer gave 4-10 bands of different molecular weights for each accession. Bands were scored as present or absent for each accession and variation among accessions was quantified using Nei's genetic distances. A dendrogram summarizing phenetic relationships among the 25 accessions was generated using the unweighted pair group method with arithmetic mean. Principal component analysis was also used to evaluate genetic similarities. Three distinct groups were identified: group 1 contains five species, group 2 contains the pinnata species, and group 3 contains the nilotica species. The analysis demonstrates that the major groups of Azolla species can be easily distinguished from one an other and, in addition, that closely related accessions within species can be identified. We further found that using 10 primers, a phylogeny that is essentially the same as that derived from 22 primers can be constructed. Our results suggest that total DNA extracted from the Azolla-Anabaena symbionts is useful for classification and phylogenetic studies of Azolla. PMID- 8405986 TI - Transcriptional and translational study of the Drosophila subobscura hsp83 gene in normal and heat-shock conditions. AB - In this paper we report a transcriptional and translational study of the hsp83 gene of Drosophila subobscura. This gene is located at the 18C region of the J chromosome. A monoclonal antibody raised against hsp83 was used for the immunological detection of this protein by Western blotting throughout the development of D. subobscura in control and heat-shock conditions. Our results indicate that puff 18C is not only heat-shock inducible but is also expressed during normal development and its level of expression increases at the end of the prepupa period. We detected hsp83 at normal temperatures, in particular developmental stages with the exception of the larval and the beginning of prepupa formation. Hsp83 was induced by heat shock in all stages studied with the exception of 1st instar larvae. We found that temperatures in excess of 26 degrees C were sufficient to induce hsp83. In addition, at temperatures from 26 to 34 degrees C, the increase in hsp83 synthesis was accompanied by increased transcription of the hsp83 gene; this positive correlation was not observed at 37 degrees C. PMID- 8405987 TI - An optimized fluorescence in situ hybridization procedure for detecting rye chromosomes in wheat. AB - In situ hybridization with an interspersed repeat clone from rye, pSc119, was shown to be useful for detecting rye chromosomes introduced into wheat. However, since pSc119 also shows strong hybridization to a few sites in certain wheat chromosomes, small rye chromosome segments added to wheat may be difficult to detect. In this study, detection of rye chromosomes present in triticale and triticale X wheat hybrids was accomplished with the use of a subfragment from pSc119 (pSc119.1) whose sequence is dispersed throughout the rye chromosomes and only weakly cross-hybridizes to a few telomeric and centromeric regions of wheat. The in situ hybridization conditions were optimized to readily distinguish rye chromosomes from wheat chromosomes without the need for intensive analysis of hybridization patterns. Rye chromosomes were readily detected using fluorescence in situ hybridization. Fluorescence detection provided increased sensitivity over enzymatic detection and allowed signals to be amplified with repeated use of biotinylated anti-avidin antibody and avidin-FITC. Detection of rye chromatin was further optimized by doubling the probe concentration. Finally, double exposure photography of the same cell with two different filters provided another means to further increase the contrast between rye and wheat chromosomes. PMID- 8405988 TI - The genetics of the Luria-Latarjet effect in bacteriophage T4: evidence for the involvement of multiple DNA repair pathways. AB - The Luria-Latarjet effect is an increase in resistance of a virus to DNA damage during infection of a host. It has often been assumed to involve recombinational repair, but this has never been demonstrated experimentally. Using nine bacteriophage (phage) T4 mutants, I present evidence indicating that, for phage T4, the Luria-Latarjet effect is due to three repair pathways-excision repair, post-replication-recombinational-repair (PRRR) and multiplicity reactivation (MR) (a second form of recombinational repair). The results also show that the Luria Latarjet effect develops in two stages. The first stage starts soon after infection. Damage which occurs during the first stage can be repaired by excision repair or PRRR. The second stage appears to start after the first round of DNA replication is complete. DNA damage which occurs during this stage can apparently be repaired by MR as well as the other two repair pathways. The results of this study support the hypothesis that recombinational repair has been selected to ensure that the progeny phage genomes which are packaged have minimum DNA damage. Since other viruses which infect bacterial, animal and plant cells show a Luria Latarjet effect similar to that in phage T4, the conclusions from this study may have wide applicability. PMID- 8405989 TI - Dosage dependent modifiers of white alleles in Drosophila melanogaster. AB - As part of a study to identify dosage-sensitive modifiers of the white eye colour locus and the retrotransposon, copia, a segmental aneuploid screen was conducted. It surveys the autosomal complement of the genome for dosage dependent modifiers of white, including ones effective upon retrotransposon insertion-induced alleles. Several regions were found which, when present as a segmental trisomy, affected one or more of the alleles tested in a strong and consistent fashion. Two of these regions have been identified as containing previously described modifiers, Darkener-of-apricot and Inverse regulator-a. The remainder identify new white allele modifiers. Selected segmental monosomics were also tested where possible for regions exhibiting a trisomic effect. At least three regions were found to have a dosage-dependent effect in one, two and three copies. PMID- 8405990 TI - Identification of ornithine decarboxylase as a trait gene for growth in replicated mouse lines divergently selected for lean body mass. AB - Studies of lines of mice selected for body mass have shown that there is a significant genetic component affecting this trait although the nature of the genes involved remains to be elucidated. Using replicate lines of mice, our studies have shown that two different variants of the mouse ornithine decarboxylase (ODCase) gene have been selected in replicate lines of mice selected for high and low lean body mass respectively. One variant is associated with an increased peak of ODCase activity in embryos (10-13 days of gestation) in all high mass lines and with a restriction fragment length polymorphism of the expressed gene. The increased ODCase activity coincides with increased ODCase mRNA levels in the high mass selected lines. These results provide evidence implicating ornithine decarboxylase as a major factor in cell growth, and as a candidate 'trait gene'. PMID- 8405991 TI - Parental origin of mutant allele does not explain absence of gene dose in X linked Hyp mice. AB - The expectation for a gene dose effect in an X-linked phenotype is that the corresponding metrical trait in heterozygous females will lie between values for affected hemizygous males and unaffected males and females. We made sequential measurements (at 30, 60, 90, 120 and 150 days) of serum phosphate concentration and tail length in mice with X-linked hypophosphatemia (genotypes: Hyp/Y, Hyp/+ and Hyp/Hyp) and in their normal litter-mates (genotypes: +/Y, +/+). We also measured renal mitochondrial 25-hydroxyvitamin D3-24-hydroxylase (24-hydroxylase) activity in 5 to 7-month-old mice fed control and low phosphate diets and representing all five genotypes. The animals were obtained by controlled breeding under uniform environmental conditions. The mutant animals all had uniformly and significantly lower serum phosphate levels, shorter tail length and higher 24 hydroxylase activity relative to unaffected litter-mates. There was no evidence of a gene dose effect because values were not significantly different among the three mutant genotypes. We also studied the influence of gamete of origin on serum phosphate, tail length and renal mitochondrial 24-hydroxylase activity in the Hyp/+ offspring of affected males (Hyp/Y) or affected females (Hyp/+ or Hyp/Hyp). We found no effect on the distribution of trait values. We conclude that parental origin of mutant allele does not explain the absence of a gene dose effect in Hyp mice. PMID- 8405992 TI - Heritabilities of growth curve parameters and age-specific expression of genetic variation under two different feeding regimes in Japanese quail (Coturnix coturnix japonica). AB - This study investigated genetic variation in growth and final size in relationship to differences in heritabilities under good and poor feeding conditions. Heritabilities of growth and final size were estimated for several traits under ad libitum and restricted feeding conditions. A 30% feed restriction from hatching to 44 days of age in Japanese quail chicks decreased body weight and tarsus length at 44 days of age and the length of the third primary convert feather at 24 days of age relative to controls fed ad libitum. Wing length at 44 days of age was not significantly different for ad libitum fed and restricted quail. Genetic variances for body weight and tarsus length were very large throughout growth which resulted in heritability estimates close to one for these traits. The genetic correlations among feeding treatments were low, indicating that different genes were affecting growth under the two treatments. Growth was described by the components: asymptote, growth period, and shape of the growth curve following the modified Richards growth curve model (Brisbin et al. 1986). Tarsus length, which had high heritability of the parameter 'growth period' of the model, tended to display a higher heritability under the restriction than under ad libitum feeding. Body weight and feather length, which had either no heritable or low heritable 'growth periods' estimates, tended to be more heritable under ad libitum feeding. The shape parameter of the growth curve was not heritable for any trait, except tarsus length under restricted feeding. PMID- 8405993 TI - Temperature related fertility selection on body size and the sex-ratio gene arrangement in Drosophila pseudoobscura. AB - We measured temperature-dependent fertility selection on body size in Drosophila pseudoobscura in the laboratory. One hundred single females of each of the three karyotypes involving the 'sex-ratio' (SR) and the standard (ST) gene arrangement on the sex chromosome laid eggs at either 18 or 24 degrees C. The experiment addressed the following hypotheses: (a) Fertility selection on body size is weaker at the higher temperature, explaining in part why genetically smaller flies appear to evolve in populations at warmer localities. (b) Homokaryotypic SR females are less fecund than homokaryotypic ST females, possibly mediated by the effect of body size on fertility, explaining the low frequencies of SR despite its strong advantage due to meiotic drive. The data were also expected to shed light on a mechanism for the evolution of plasticity of body size through fertility selection in environments with an unpredictable temperature regime. Hypothesis (a) was clearly refuted because phenotypically larger ST females had an even larger fertility surplus at the higher temperature and, more importantly, the genetic correlation between fertility and body size disappeared at the lower temperature. As to (b), we found that temperature affects fertility directly and indirectly through body size such that ST and SR females were about equally fecund at both temperatures, although different in size and size-adjusted fertility. We observed heterosis for both size and fertility, which might stabilize the polymorphism in nature. The reaction norms of body size to the temperature difference were steeper for ST females than for SR females, implying that fertility selection could change phenotypic plasticity of body size in a population. Selection on body size depended not only on the temperature, but also on the karyotypes, suggesting that models of phenotype evolution using purely phenotypic fitness functions may often be inadequate. PMID- 8405994 TI - Cooperative DNA binding of p53 with TFIID (TBP): a possible mechanism for transcriptional activation. AB - The p53 tumor-suppressor gene product, a sequence-specific DNA-binding protein, has been shown to act both as a transcriptional activator and repressor in vivo and in vitro. Consistent with its roles in regulating transcription are recent observations that p53 binds directly to the TATA box-binding protein (TBP) subunit of the basal transcription factor TFIID. Here, we show that p53 cooperates with either recombinant TBP or partially purified TFIID in binding to a DNA fragment containing both a specific p53-binding site (RGC) and a TATA box (RGC-TATA). Surprisingly, both TBP and TFIID also stimulate p53 binding to DNA containing a specific p53-binding site but lacking a TATA box. These data are supported by the observation that p53 and Drosophila TBP combinatorily activate transcription in vivo. Our results suggest that p53 activates transcription through the formation of a more stable p53-TFIID-promoter complex. We also examined whether p53 might affect the ability of TBP or TFIID to interact with DNA containing a TATA box but lacking a p53-binding site. Although p53 strongly inhibited the interaction of TBP with such DNA, it had virtually no effect on TFIID binding. Thus, transcriptional repression by p53 may require additional functions other than inhibiting TBP binding. PMID- 8405995 TI - Heterodimerization of the transcription factors E2F-1 and DP-1 leads to cooperative trans-activation. AB - The E2F transcription factor has been implicated in the regulation of genes whose products are involved in cell proliferation. Two proteins have recently been identified with E2F-like properties. One of these proteins, E2F-1, has been shown to mediate E2F-dependent trans-activation and to bind the hypophosphorylated form of the retinoblastoma protein (pRB). The other protein, murine DP-1, was purified from an E2F DNA-affinity column, and it was subsequently shown to bind the consensus E2F DNA-binding site. To study a possible interaction between E2F-1 and DP-1, we have now isolated a cDNA for the human homolog of DP-1. Human DP-1 and E2F-1 associate both in vivo and in vitro, and this interaction leads to enhanced binding to E2F DNA-binding sites. The association of E2F-1 and DP-1 leads to cooperative activation of an E2F-responsive promoter. Finally, we demonstrate that E2F-1 and DP-1 association is required for stable interaction with pRB in vivo and that trans-activation by E2F-1/DP-1 heterodimers is inhibited by pRB. We suggest that "E2F" is the activity that is formed when an E2F-1-related protein and a DP-1-related protein dimerize. PMID- 8405996 TI - Circadian transcription of the cholesterol 7 alpha hydroxylase gene may involve the liver-enriched bZIP protein DBP. AB - The liver-enriched transcription factor DBP is expressed with a stringent circadian rhythm. We present evidence that DBP is a regulator of the circadian expression of the rat gene encoding cholesterol 7 alpha hydroxylase (C7 alpha H), the rate-limiting enzyme in the conversion of cholesterol to bile acids. As with DBP, C7 alpha H mRNA reaches peak levels in the evening, and its cycling is independent of daily food and light cues. As predicted for a DBP target gene, the primary level of C7 alpha H circadian expression is at the transcriptional level. DBP can activate the C7 alpha H promoter in cotransfection assays through a cognate DNA site centered around -225. In nuclear extracts prepared by a novel method that, in contrast to conventional techniques, yields near-quantitative recovery of DBP and other non-histone proteins, the DNA site required for DBP activation is the predominant site of occupancy by nuclear factors on the C7 alpha H promoter. At this site, the predominant binding activity is an evening specific complex of which DBP is a component. These data suggest that DBP may play an important role in cholesterol homeostasis through circadian transcriptional regulation of cholesterol 7 alpha hydroxylase. PMID- 8405997 TI - Multiple cis-acting signals for export of pre-U1 snRNA from the nucleus. AB - We have identified cis-acting sequences that promote nuclear export of pre-U1 RNA injected into Xenopus oocyte nuclei. At least three elements, the 5' m7G cap, the 3'-terminal stem-loop structure, and sequences in the 5'-terminal 124 nucleotides, contribute to efficient export of this RNA. Both the 5' and 3' export signals can function separately and do so independently of the cap structure. Experiments using hybrid RNAs indicate that the 5' and 3' export sequences of U1 RNA are sufficient to direct export of the heterologous, otherwise nonexportable, U6 RNA. The absence of comparable export signals in U6 RNA appears to be responsible for its retention in the nucleus. Stability of the pre-snRNAs in the nucleus depends on the presence of both a 5' cap structure and a 3' base-paired stem. The 5' m7G cap is neither sufficient nor necessary for nuclear export. The m7G cap by itself did not promote export of U6 RNA or nonspecific small RNAs. Moreover, substitution of this cap with either an AppG cap or gamma-mppG cap did not eliminate export of either full-length or a "minimal" U1 RNA (lacking most of the internal U1 RNA sequences), but it reduced the rate of export by about two to threefold. However, in the absence of the 3' stem-loop, substitution of the m7G cap led to a greater decrease in export rate, underscoring the cooperative action of the three different export elements of pre U1 RNA. The m7G cap analog, m7GpppG, selectively destabilized pre-U1 RNA within the nucleus. Thus, nuclear components that recognize the 5' m7G cap may be important for both the stability and the export of pre-U1 RNA. PMID- 8405998 TI - Four yeast spliceosomal proteins (PRP5, PRP9, PRP11, and PRP21) interact to promote U2 snRNP binding to pre-mRNA. AB - We have analyzed the functions of several pre-mRNA processing (PRP) proteins in yeast spliceosome formation. Here, we show that PRP5 (a DEAD box helicase-like protein), PRP9, and PRP11 are each required for the U2 snRNP to bind to the pre spliceosome during spliceosome assembly in vitro. Genetic analyses of their functions suggest that they and another protein, PRP21, act concertedly and/or interact physically with each other and with the stem-loop IIa of U2 snRNA to bind U2 snRNP to the pre-mRNA. Biochemical complementation experiments also indicate that the PRP9 and PRP11 proteins interact. The PRP9 and PRP11 proteins may be functioning similarly in yeast and mammalian cells. The requirement for ATP and the helicase-like PRP5 protein suggests that these factors might promote a conformational change (involving either the U1 or U2 snRNP) that is required for the association of U2 snRNP with the pre-mRNA. PMID- 8405999 TI - Mutation of an RSV intronic element abolishes both U11/U12 snRNP binding and negative regulation of splicing. AB - A cis-acting negative regulator of splicing (NRS) within the gag gene of RSV is involved in control of the relative levels of spliced and unspliced viral mRNAs. Insertion of the NRS into the intron of an adenovirus pre-mRNA resulted in inhibition of splicing in vitro before the first cleavage step. Analyses of spliceosome assembly with this substrate showed that it formed large RNP complexes that did not migrate like mature spliceosomes on native gels. Affinity selection of the RNP complexes formed on NRS-containing pre-mRNAs showed an association with U11 and U12 snRNPs, as well as with the spliceosomal snRNPs. Immunoprecipitation with antisera specific for U1 and U2 snRNPS showed binding of both snRNPs to NRS RNA. A 7-nucleotide missense mutation in the NRS that prevented binding of U11 and U12 snRNPs impaired NRS activity in vivo, suggesting a functional role for U11 and U12 snRNPs in the inhibition of splicing mediated by the RSV NRS RNA. PMID- 8406000 TI - Establishment of ventral cell fates in the Drosophila embryonic ectoderm requires DER, the EGF receptor homolog. AB - The embryonic ectoderm in Drosophila displays a highly organized arrangement of specific structures along the dorsal-ventral axis. To establish this characteristic design, cells must receive instructive cues regarding their position. We present evidence that during stages 8-9 of embryonic development, the Drosophila EGF receptor homolog (DER) is essential for determining the identity of cells within the ventral ectoderm. In the absence of DER activity at this phase, alterations in cell fate are observed: Ventral cells acquire more dorsal fates, as visualized by the expression profile of specific markers. The ventralizing effect of DER appears to function later than that of the dorsalizing dpp pathway, and the spatial overlap between them is minimal. A model for the determination of cell fates along the dorsal-ventral axis involving the two pathways is presented. Some aspects of the mutant ectodermal and CNS phenotypes of the DER locus (faint little ball, flb) resemble the phenotype of mutations from the spitz group. Synergistic interactions between flb and spitz or Star mutations suggest that these genes participate in a common signaling pathway. PMID- 8406001 TI - Antineurogenic phenotypes induced by truncated Notch proteins indicate a role in signal transduction and may point to a novel function for Notch in nuclei. AB - Loss of any one of several neurogenic genes of Drosophila results in overproduction of embryonic neuroblasts at the expense of epidermoblasts. In this paper a variety of altered Notch proteins are expressed in transgenic flies. Dominant lethal, antineurogenic phenotypes were produced by expression of three classes of mutant proteins: (1) a protein comprised of the cytoplasmic domain of Notch and devoid of sequences permitting membrane association; (2) a transmembrane protein lacking the extracellular, lin12/Notch repeats; and (3) transmembrane proteins carrying amino acid substitutions replacing one or both extracellular cysteines thought to be involved in Notch dimerization. These Notch proteins not only suppress the neural hypertrophy observed in Notch- embryos, but also generate a phenotype in which elements of the embryonic nervous system are underproduced. Action of the intracellular cdc10 repeats appears to be essential for wild-type Notch function or for the antineurogenic activity of these proteins. The activities of the dominant, gain-of-function proteins indicate that Notch functions as a signal transducing receptor during ectoderm development. Production of antineurogenic Notch proteins in embryos deficient for the other neurogenic genes allowed functional dependencies to be established. Delta, mastermind, bigbrain, and neuralized appear to function in elaboration of a signal upstream of Notch. Genes of the Enhancer of split complex act after Notch. The cytoplasmic domain of Notch contains nuclear localization sequences that function in cultured cells, and one of the Notch antineurogenic proteins, the cytoplasmic domain, accumulates in nuclei in vivo. PMID- 8406002 TI - Spatial and temporal phosphorylation of a transcriptional activator regulates pole-specific gene expression in Caulobacter. AB - Polar localization of proteins in the Caulobacter predivisional cell results in the formation of two distinct progeny cells, a motile swarmer cell and a sessile stalked cell. The transcription of several flagellar promoters is localized to the swarmer pole of the predivisional cell. We present evidence that the product of the flbD gene is the transcriptional activator of these promoters. We show that FlbD is distributed in all cell types and in both poles of the predivisional cell. We also demonstrate that FlbD can be phosphorylated, and that a FlbD kinase activity is under cell cycle control. Cells expressing a FlbD mutant that should activate transcription in the absence of phosphorylation, exhibited an alteration in the temporal pattern of flagellin transcription. Furthermore, predivisional cells expressing the mutant FlbD failed to polarly localize flagellin synthesis. We propose that the phosphorylation of FlbD is restricted to the swarmer compartment of the predivisional cell, and serves as the control point for regulating the spatial transcription of flagellar promoters. PMID- 8406003 TI - The activity of the Escherichia coli transcription factor FNR is regulated by a change in oligomeric state. AB - The transcription factor FNR globally regulates gene expression in response to oxygen deprivation in Escherichia coli. To understand how oxygen deprivation activates FNR, a constitutively active FNR* mutant protein, DA154, was studied to determine how this mutant bypassed the normal regulation pathway. When purified from aerobically grown cells, the DA154 protein had a larger apparent native molecular mass and an increased affinity for a consensus FNR target site as compared with wild-type FNR prepared under identical conditions. These results suggested that FNR* DA154 may bypass the normal regulation pathway by converting FNR from an inactive monomer to an active dimer under aerobic conditions. To determine whether wild-type FNR is active as a dimer under anaerobic conditions, FNR mutants were isolated that inhibit the activity of wild-type FNR by forming mixed dimers (i.e., dominant-negative mutants). These dominant-negative FNR mutants were shown to have substitutions in the putative DNA-binding domain and to be defective in binding to a consensus FNR DNA target site in vitro. One representative dominant-negative mutant, EK209, was also shown to be unable to form mixed oligomers in vivo under aerobic conditions, suggesting that FNR may be monomeric in the inactive state. Taken together, these data have led us to propose that under anaerobic conditions FNR is a dimer that is active for DNA binding, and under aerobic conditions, FNR is inactivated by conversion to a monomer. PMID- 8406004 TI - Ets-related protein PU.1 regulates expression of the immunoglobulin J-chain gene through a novel Ets-binding element. AB - In a primary immune response, a signal from interleukin-2 (IL-2) activates transcription of the gene encoding the pentamer IgM joining component, the J chain. Recently, a bifunctional control element (JB) in the J-chain promoter has been identified. This finding was pursued in the present study by purifying and characterizing the nuclear protein (NF-JB) that mediates the positive regulatory activity of the JB element. The analyses revealed that NF-JB is identical to the Ets-related B-cell- and macrophage-specific transcriptional factor, PU.1, despite the fact that the JB site lacks the GGA core reported to be essential for binding by members of the Ets oncoprotein family. The two factors were found to be indistinguishable with respect to their DNA-binding characteristics, size, and peptide structure. Moreover, in transient transfection assays, PU.1 alone activated reporter constructs containing the JB cis-element, and the activation was shown to be dependent on a glutamine-rich sequence in the amino-terminal portion of PU.1. Finally, a dominant negative mutant of PU.1 was capable of suppressing the transcriptional activity of a 1.2-kb J-chain promoter sequence. These results establish an important role for PU.1 in the regulation of immunoglobulin J-chain gene expression and provide new insights into the function(s) of the Ets transcription factors in lymphoid cells. PMID- 8406005 TI - The immunoglobulin mu enhancer core establishes local factor access in nuclear chromatin independent of transcriptional stimulation. AB - Factor access in chromatin has been proposed to be facilitated by transcriptional enhancers. With the aim of uncoupling factor access from transcriptional stimulation by protein-protein contacts, we analyzed the potential of enhancer fragments to confer accessibility upon a linked promoter for prokaryotic T7 RNA polymerase. Access to the T7 promoter in pre-B cells from transgenic mice was examined by transcribing chromatin of isolated nuclei with T7 RNA polymerase. A 95-bp immunoglobulin mu enhancer core element was necessary and sufficient to confer accessibility upon the T7 promoter independent of its chromosomal position. This enhancer-dependent factor access could be uncoupled from an active transcriptional state of the transgene and was not accompanied by the formation of pronounced DNase I hypersensitive sites. Additional mu enhancer sequences comprising previously identified matrix attachment regions and a cryptic promoter were required to induce DNase I hypersensitivity. Together, these data provide evidence that the 95-bp mu enhancer core can establish localized factor access in nuclear chromatin independent of detectable transcription by endogenous polymerases and suggest that multiple steps are involved in the alteration of chromatin structure. PMID- 8406006 TI - Replication-coupled chromatin assembly is required for the repression of basal transcription in vivo. AB - The chromatin assembly process coupled to DNA synthesis in the Xenopus oocyte nucleus is significantly more repressive toward basal transcription than chromatin assembly on duplex DNA. We show that chromatin assembly concurrent with DNA synthesis over the promoter region itself is causal for repression. However, the trans-activator Gal4-VP16 both relieves repression and activates transcription regardless of the chromatin assembly pathway. This activation is independent of whether Gal4-VP16 addition occurs before or after chromatin assembly. We propose that replication-coupled chromatin assembly represents a general mechanism to direct the efficient repression of basal transcription. However transcription induction by a specific activator, Gal4-VP16, occurs independent of this chromatin-mediated repression. PMID- 8406007 TI - DNA sequence recognition by Pax proteins: bipartite structure of the paired domain and its binding site. AB - Previous DNA-binding studies indicated that an intact paired domain is required for interaction of the transcription factor BSAP (Pax-5) with DNA. We have now identified a subset of BSAP recognition sequences that also bind to a truncated BSAP peptide lacking 36 carboxy-terminal amino acids of the paired domain. Sequence comparison of this class of BSAP-binding sites made it possible to unequivocally align all known BSAP-binding sites and to deduce a consensus sequence consisting of two distinct half sites. We propose here a model for the paired domain--DNA interaction in which the paired domain is composed of two subdomains that bind to the two half-sites in adjacent major grooves on the same side of the DNA helix. The existence of these half sites and of the two paired domain subregions was directly demonstrated by methylation interference analysis and by in vitro mutagenesis of both the paired domain and its recognition sequence. Both half-sites contribute to the overall affinity of a given BSAP binding site according to their match with the consensus sequence. However, none of the naturally occurring BSAP-binding sites completely conform to the consensus sequence. Instead, they contain compensatory base changes in their half-sites that explain the versatile and seemingly degenerate DNA sequence recognition of Pax proteins. Domain swap experiments between BSAP and Pax-1 demonstrated that the sequence specificity of the BSAP paired domain is determined by both its amino- and carboxy-terminal subdomains. Moreover, mutations affecting only one of the two subdomains restricted the sequence specificity of the paired domain. Such mutations have been shown previously to be the cause of mouse developmental mutants (undulated, Splotch, and Small eye) and human syndromes (Waardenburg's syndrome and aniridia) and may thus differentially affect the regulation of target genes by the mutated Pax protein. PMID- 8406008 TI - Binding of tissue-specific forms of alpha A-CRYBP1 to their regulatory sequence in the mouse alpha A-crystallin-encoding gene: double-label immunoblotting of UV crosslinked complexes. AB - The alpha A-CRYBP1 regulatory sequence (alpha A-CRYBP1RS), at nucleotides -66 to 57 of the mouse alpha A-crystallin-encoding gene (alpha A-CRY) promoter, is an important control element involved in the regulation of mouse alpha A-CRY expression. The gene encoding a protein (alpha A-CRYBP1) that specifically binds to the alpha A-CRYBP1RS sequence has been cloned from a cultured mouse lens cell line. In the present study, we have used an antibody (specific to the alpha A CRYBP1 protein and made against a synthetic peptide) to directly identify UV crosslinked protein-DNA complexes via a double-label immunoblotting technique. Multiple alpha A-CRYB1 antigenically related proteins interacted with alpha A CRYBP1RS in nuclear extracts from both a cloned mouse lens cell line (alpha TN4 1) that expresses alpha A-CRY and a mouse fibroblast line (L929) that does not express the gene. Two sizes (50 kDa and 90 kDa) of proteins reacting with the alpha A-CRYBP1-specific Ab were detected in both cell lines and, in addition, a > 200-kDa protein reacting with the Ab was unique to the fibroblast line. Thus, alpha A-CRYBP1 antigenically related proteins interact with alpha A-CRYBP1RS regardless of alpha A-CRY expression. Moreover, differential processing of the alpha A-CRYBP1 protein and/or alternative splicing of the alpha A-CRY transcript may affect expression of alpha A-CRY. PMID- 8406009 TI - A Caenorhabditis elegans act-4::lacZ fusion: use as a transformation marker and analysis of tissue-specific expression. AB - A plasmid vector that serves as a dominant marker for isolating transformed animals in Caenorhabditis elegans has been constructed as a translational fusion of the C. elegans act-4 gene (encoding actin) and the Escherichia coli lacZ gene. This gene fusion can be used as a marker in transformation rescue experiments in any fertile strain of C. elegans. Progeny of animals injected with the act 4::lacZ fusion vector are stained histochemically with XGal, and transformants turn blue. The internal eggs of stained animals remain viable, allowing recovery of the transformed strain. When the act-4::lacZ vector is co-injected with an unselected plasmid with which it shares some sequence homology, most transformants that are recovered by screening for expression of the act-4::lacZ fusion contain both plasmids. Production of active beta Gal in animals transformed with the act-4::lacZ gene fusions appears to be limited to certain tissues. A chimeric gene that contains the 5' and 3' regions of act-4 is expressed strongly in the body-wall muscles, vulval muscles, and spermathecae. Addition of the internal portion of act-4, including the protein-coding region and introns, to this chimeric gene leads to additional lacZ expression in the pharynx. PMID- 8406010 TI - Differential expression of transcripts from syb, a Drosophila melanogaster gene encoding VAMP (synaptobrevin) that is abundant in non-neuronal cells. AB - VAMP (synaptobrevin) is a highly conserved membrane protein originally described as a component of brain synaptic vesicles. The Drosophila melanogaster VAMP encoding gene (syb) comprises five exons. Splicing exons 1,2,3,4,5 (syb-b) results in a protein with a C-terminal hydrophobic domain and a negligible intraluminal domain. Splicing exons 1,2,3,5 (syb-a) predicts a protein with a 20 amino-acid luminal domain at the C terminus. The ratio of syb-a to syb-b transcripts is highly regulated during development. The syb transcripts show no enrichment in the nervous system and are present in very early embryos, well before neurogenesis. The greatest concentration of syb transcripts was found in cells of the gut and malpighian tubules. Thus, syb may have a general role in membrane trafficking and, perhaps, a role in the secretion of digestive enzymes. PMID- 8406011 TI - Presence of particular transcription regulatory elements in the 5'-intergenic region shared by the chicken H2A-III and H2B-V pair. AB - The two chicken histone gene families, H2A and H2B, contain nine and eight members, respectively, within two major histone gene clusters. Six genes each from families H2A and H2B have been found to be closely associated in inverted directions as H2A/H2B gene pairs. Two previously sequenced H2A members (H2A-I and H2A-II) encode the same amino acid (aa) sequence (class I), whereas seven sequenced H2B genes encode three different variants (classes I, II and III). In this study, we first sequenced H2A-III, a member of the H2A family, which is located in inverted orientation and 350 bp upstream from H2B-V, encoding the class-III H2B protein. The protein encoded by H2A-III differs from the class-I H2A protein in a single aa (Ala70-->Pro; class II). As a step toward elucidation of the transcriptional regulation of the H2A and H2B families, we fused this 5' intergenic region to the cat gene in inverted orientations to generate two chimeric plasmids, pH2A-III-350 and pH2B-V-350. Transient CAT assays using these constructs indicated that the promoter of H2B-V is more active than that of H2A III. CAT assays with 5'-deletion mutants of H2A-III and H2B-V showed that they each possess particular transcriptional motifs which are located relatively close to, or apart from, their own coding regions. These findings, together with those reported previously on the H2A-V/H2B-II pair, suggest distinct manners of transcription regulation of different members of the chicken histone gene families, H2A and H2B. PMID- 8406012 TI - Transcriptional analyses of the gene region that encodes human histidyl-tRNA synthetase: identification of a novel bidirectional regulatory element. AB - A recombinant phage clone containing the 5' end of the gene HRS encoding human histidyl-tRNA synthetase (HRS) has been isolated. Primer extension analyses indicated that there are two types of HRS transcripts. The longer transcripts were initiated from a single transcription start point (tsp) located approximately 455 bp upstream and the shorter transcripts were initiated from multiple tsp located approximately 38 to 82 bp upstream from the HRS ATG start codon. Functionally, we have identified two regions (+1 to -122; -185 to -502), each of which when placed 5' of a promoterless cat construct can initiate transcription in both orientations after transfection into HeLa cells. A pair of imperfect inverted repeats (IIR) was located within the region +1 to -122. Using mobility shift assays, we have identified a nuclear factor that binds specifically to each half of the IIR. However, this pair of IIR (-73 to -110) was not sufficient for bidirectional transcription activity. At least one copy of a 27-bp oligodeoxyribonucleotide (oligo), which spans -94 to -120, was required in order to facilitate bidirectional transcription activity. From mobility shift assays using HeLa cell nuclear extracts and this 27-bp oligo, we have identified two DNA-protein complexes, both of which are presumably required to initiate bidirectional transcription. PMID- 8406013 TI - Identification of novel sequences in the repertoire of hypervariable TRE17 genes from immortalized nonmalignant and malignant human keratinocytes. AB - The TRE17 oncogene, originally cloned from transfected DNA of Ewing's sarcoma cells, maps to chromosome 17q and is expressed in a wide variety of human cancer cells. We recently detected the variants of this gene by using the polymerase chain reaction (PCR) and sequencing from the first exon to the third intron. Based on sequence homology scores, the variants could be grouped into three families, denoted alpha, beta, and gamma. Here, we used human keratinocytes from healthy skin which had been spontaneously immortalized and then rendered malignant by serum privation in vitro. Both immortalized and malignant cells expressed TRE17 sequences to the same extent, and, according to the restriction site analysis of cloned PCR products, both contained common and rare TRE17 variants in similar proportions. These variants, one of each from both cell types, were then sequenced and compared with those from the previous study. In the phylogenetic tree, they clustered with alpha and gamma at the most distant tree positions. The overall fraction of conserved sites in the whole TRE17 repertoire was 80%. An unexpected feature of the observed variability was that intronic sites were significantly better conserved than exonic sites. Members of TRE17 gamma detected in immortalized and malignant keratinocytes differed one from another, and both differed from the TRE17 gamma already identified in Ewing's sarcoma. No TRE17 gamma has been found so far in healthy tissues, thus leaving open the possibility of its origin from TRE17 beta by somatic changes during tumor progression. PMID- 8406014 TI - Zea mI, the maize homolog of the allergen-encoding Lol pI gene of rye grass. AB - Sequence analysis of a pollen-specific cDNA from maize has identified a homolog (Zea mI) of the gene (Lol pI) encoding the major allergen of rye-grass pollen. The protein encoded by the partial cDNA sequence is 59.3% identical and 72.7% similar to the comparable region of the reported amino acid sequence of Lol pIA. Southern analysis indicates that this cDNA represents a member of a small multigene family in maize. Northern analysis shows expression only in pollen, not in vegetative or female floral tissues. The timing of expression is developmentally regulated, occurring at a low level prior to the first pollen mitosis and at a high level after this postmeiotic division. Western analysis detects a protein in maize pollen lysates using polyclonal antiserum and monoclonal antibodies directed against purified Lolium perenne allergen. PMID- 8406015 TI - Specific interaction between adenoviral 55-kDa E1B protein and in vivo produced p53 fusion proteins. AB - Several protein fusion systems have been used in recent years to study protein protein and DNA-protein interactions. Most of them use bacterially produced proteins which have several inherent disadvantages, notably, the absence of correct post-translational modifications and the frequent insolubility of recombinant proteins. We sought to develop a system to study proteins interacting with the nuclear phosphoprotein p53, which is believed to be a tumor suppressor. To prepare fusions of p53, we developed a convenient system that permits both in vivo and in vitro production and easy affinity purification of peptides and protein fragments as glutathione-transferase fusions. We placed the coding sequence of the Schistosoma japonica glutathione S-transferase (GST) under the control of the strong CMV/T7 promoter and SV40 splice and polyadenylation signals. An extensive polylinker (MCS) at the 3' end of the GST gene is preceded by the sequence encoding the cleavage site of the site-specific protease. We cloned the complete coding sequences of human wild-type p53, as well as p53 mutants representing all four mutational hotspots (codons 141, 175, 248, and 273), into our expression vector. In vitro transcription using the upstream T7 promoter and translation in reticulocyte lysates form an easy way to produce hybrid proteins; affinity purification on a glutathione-agarose column removes proteins that are present in reticulocyte lysates. We have also studied specific in vivo interactions of human p53 with the adenoviral 55-kDa E1B protein by transfecting expression constructs of GST-p53 fusions into human Ad5-transformed 293 cells. PMID- 8406016 TI - A repetitive element in the genome of Atlantic salmon, Salmo salar. AB - When Atlantic salmon (Salmo salar) genomic DNA is digested with the restriction endonuclease BglI and the fragments separated by agarose-gel electrophoresis, bands corresponding to approximately 430 and 923 bp are visualized after EtdBr staining. The 923-bp band was excised from a preparative gel and used to screen a salmon genomic library for recombinant phage (re-phage) containing the repeat. The BglI repeat element is tandemly arrayed, and an array from one re-phage has been sequenced. The BglI repeats comprise 2.3% of the S. salar genome and have been found in the vicinity of rDNA genes (encoding ribosomal RNA). Southern blot hybridization detects a homologue of the Atlantic salmon BglI repeat in the brown trout (Salmo trutta) genome, but not in other salmonids. However, a DNA fragment with sequence homology to part of the BglI repeat has recently been isolated from Arctic charr (Salvelinus alpinus; S.E. Hartley and W.S.D., unpublished data). In addition, the BglI repeat detects RFLPs in Atlantic salmon. PMID- 8406017 TI - Isolation and characterization of the gene encoding the rat alpha 1B adrenergic receptor. AB - Using a rat alpha 1B adrenergic receptor(AR)-encoding cDNA probe, we isolated two genomic clones from a rat liver genomic DNA library. Southern blot analysis and nucleotide sequencing indicate that the rat alpha 1B AR gene has two exons and a single large intron of at least 16 kb. Analysis of the sequence of the 5' flanking region suggests that this gene has the features of a housekeeping gene: it has neither a TATA box nor a CAAT box, but has multiple transcription start points (tsp), multiple Sp1-binding sites, and a high G + C content. The 5' flanking region also contains consensus sequences for AP1- and AP2-binding sites, and putative sites for cyclic AMP (CRE), glucocorticoid (GRE), and thyroid responses (TRE). The 3'-flanking region contains a putative polyadenylation signal (ATTAAA) 492 bp downstream from the stop codon. A comparison of the rat alpha 1B AR sequence with that of the recently cloned human alpha 1B AR gene showed that the overall structure of this gene is highly conserved, with some notable differences in the 5'-flanking region. PMID- 8406018 TI - Characterization of the genes encoding carbonic anhydrase I of chimpanzee and gorilla: comparative analysis of 5' flanking erythroid-specific promoter sequences. AB - The genes encoding carbonic anhydrase I (CA I) have been characterized for chimpanzee (Pan troglodytes) and gorilla (Gorilla gorilla). In addition, 44 nucleotides (nt) at the 5' end of the noncoding first exon (exon 1a), which is unique to the erythroid CA I mRNA, together with 188 nt of the adjacent 5' flanking regions, were sequenced for the corresponding positions of the CA I of orangutan, pigtail macaque, and squirrel monkey. When these 5' flanking regions are compared, along with those published for human and mouse CA I, they were found to contain several conserved sequences that may bind factors involved in the erythroid-specific expression of CA I. Comparisons of the human, chimpanzee, and gorilla coding and noncoding CA I sequences do not significantly deviate from a pattern of trichotomy for the evolutionary origins of these three hominoid species. PMID- 8406019 TI - A phage T7 class-III promoter functions as a polymerase II promoter in mammalian cells. AB - A phage T7 class-III promoter (pT7), which is highly specific for T7 RNA polymerase in bacteria, was tested in mammalian cells for its specificity. After having shown that T7 RNA polymerase can transcribe from pT7 in the nucleus of stably transformed cells [Lieber et al., Nucleic Acids Res. 17 (1989) 8485-8493], we describe here that pT7 could also direct efficient intracellular gene expression in the absence of T7 RNA polymerase. Using the genomic human growth hormone-encoding gene and the firefly luciferase-encoding gene as reporters, we found expression levels comparable with those obtained with the Rous sarcoma viral promoter. Inhibition of expression with alpha-amanitin suggests that transcription is by RNA polymerase II. Binding studies with HeLa cell extracts clearly show that synthetic pT7 sequences are specifically bound (gel retardation) and that the promoter region is protected from DNase degradation. The experimental data, as well as the nucleotide sequence, suggest that pT7 has properties of an initiator element. Indeed, the activity of pT7 can be stimulated by the presence of an upstream element or an enhancer. These results have practical implications for the use of pT7 in mammalian expression vectors. Commercial pT7 plasmids can be used for both prokaryotic and eukaryotic expression systems. PMID- 8406020 TI - Differential secretion and glycosylation of recombinant human chorionic gonadotropin (beta hCG) synthesized using different promoters in the baculovirus expression vector system. AB - Recombinant baculoviruses vAc beta hCGCOR and vAc beta hCGPOL, carrying the gene (beta hCG) encoding the beta-subunit of human chorionic gonadotropin under the transcriptional control of the late AcNPV core protein gene promoter (PCOR) and the very late polyhedrin gene promoter (PPOL), respectively, were constructed and used to infect lepidopteran cells. Western blot analysis of intra- and extracellular recombinant beta hCG (re-beta hCG) revealed that the secretion of beta hCGCOR was relatively higher. Enzymatic and chemical analysis of carbohydrates showed that beta hCGCOR was more glycosylated than beta hCGPOL. However, the insect-derived beta hCG, with a high-mannose type of sugar, was glycosylated differently and to a lesser extent when compared with the native, urinary beta hCG, and consequently, beta hCGCOR was more bioactive on a unit-mass basis than beta hCGPOL. This temporal gene expression strategy, besides being able to circumvent the 'secretory load' encountered during the synthesis of extensively glycosylated proteins in the baculovirus system, also offers a model to study the role of carbohydrates, both in qualitative and quantitative terms, in protein structure and function. PMID- 8406021 TI - Characterisation of two identical independent non-homologous integration sites in mouse embryonic stem cells. AB - On analysis of 46 Geneticin-resistant (GtR) cell lines, derived by electroporation of mouse embryonic stem (ES) cells with a promoterless neo vector, we observed that in two independently derived cell lines, the vector had integrated into the same locus. The sequence flanking the vector integration site in both cell lines was cloned and sequenced. The vector had integrated into a 3 to 6-bp region in both cell lines. No homology is observed between the integration site sequence and the vector sequence. PMID- 8406022 TI - Deletion of the beta-turn/alpha-helix motif at the exon 2/3 boundary of human c Myc leads to the loss of its immortalizing function. AB - The protein product (c-Myc) of the human c-myc proto-oncogene carries a beta turn/alpha-helix motif at the exon2/exon3 boundary. The amino acid (aa) sequence and secondary structure of this motif are highly conserved among several nuclearly localized oncogene products, c-Myc, N-Myc, c-Fos, SV40 large T and adenovirus (Ad) Ela. Removal of this region from Ad E1a results in the loss of the transforming properties of the virus without destroying its known transregulatory functions. In order to analyse whether deletion of the above mentioned region from c-Myc has a similar effect on its transformation activity, we constructed a deletion mutant (c-myc delta) lacking the respective aa at the exon2/exon3 boundary. In contrast to the c-myc wild-type gene product, constitutive expression of c-myc delta does not lead to the immortalization of primary mouse embryo fibroblast cells (MEF cells). This result indicates that c Myc and Ad El a share a common domain which is involved in the transformation process by both oncogenes. PMID- 8406023 TI - The human pancreatic lipase-encoding gene: structure and conservation of an Alu sequence in the lipase gene family. AB - The isolation and characterization of the human gene (hPL) encoding pancreatic lipase is reported. The gene has 13 exons dispersed in about 20 kb of genomic DNA. A pseudogene of hPL was also partially characterized. An Alu sequence is conserved in the homologous introns of hPL and the lipoprotein lipase-encoding gene. PMID- 8406024 TI - Cloning and expression in Escherichia coli of the cDNA encoding human cardiac troponin I. AB - We have cloned and sequenced the human cardiac troponin I (cTnI)-encoding cDNA with the aim of expressing the cDNA in Escherichia coli. The cDNA was successfully expressed as a fusion product with beta-galactosidase and as an unfused protein. Both polypeptides were recognised by an anti-human cTnI antibody. PMID- 8406025 TI - Cloning and sequence analysis of cDNAs encoding human hippocampus N-methyl-D aspartate receptor subunits: evidence for alternative RNA splicing. AB - Several cDNA clones encoding human N-methyl-D-aspartate receptor (hNR1) subunit polypeptides were isolated from a human hippocampus library. Degenerate oligodeoxyribonucleotide (oligo) primers based on the published rat NR1 (rNR1) amino acid (aa) sequence [K. Moriyoshi et al. Nature 354 (1991) 31-37] amplified a 0.7-kb fragment from a human hippocampus cDNA library, via the polymerase chain reaction (PCR). This fragment was used as a probe for subsequent hybridization screening. DNA sequence analysis of 28 plaque-purified clones indicated three distinct classes, designated hNR1-1, hNR1-2 and hNR1-3, presumably generated by alternative RNA splicing. One of these clones, hNR1-1(5A), was isolated as a full length cDNA. The hNR1-2 and hNR1-3 cDNAs represented 66.8 and 98.9%, respectively, of the total aa coding information predicted for the polypeptides. The hNR1 cDNAs demonstrated an 84-90.8% nucleotide (nt) identity with the corresponding rodent cDNAs. The nt sequences of hNR1-1, hNR1-2 and hNR1-3 would encode 885-, 901- and 938-aa proteins, respectively, that have 99.1-99.8% identity with the corresponding rodent NR1 (roNR1) subunits. The changes between the predicted aa sequences of hNR1 and the corresponding roNR1 subunits are confined to the extracellular N-terminal regions. We have also identified two possible allelic variations of the hNR1-3 cDNA that result in aa substitutions in the extracellular N- and C-terminal regions. One of these naturally occurring aa variations is situated within a potential glutamate-binding site. PMID- 8406026 TI - Sequence of a cDNA encoding wheat eukaryotic protein synthesis initiation factor 4A. AB - A cDNA encoding wheat translation initiation factor 4A (eIF-4A) was isolated from a wheat cDNA library and sequenced. The deduced wheat eIF-4A amino acid sequence from the cDNA is compared to eIF-4A from Arabidopsis thaliana, tobacco, mouse and Saccharomyces cerevisiae. Putative RNA helicase motifs, and putative ATP-binding and hydrolysis sites are identified. PMID- 8406027 TI - Sequence of manganese superoxide dismutase-encoding cDNAs from multiple mouse organs. AB - Manganese superoxide dismutase (MnSOD)-encoding cDNAs from multiple mouse organs, including liver, kidney, brain, spleen and heart, show three sequence differences compared with the previously published mouse placenta MnSOD cDNA. The differences cause substitutions of Val(-7)-->Gly (GTG-->GGT) and Met(114)-->Val (ATG-->GTG) and a G746 deletion in the 3' untranslated region. The analysis, by reverse transcriptase-polymerase chain reaction-direct sequencing, of BALB/c mouse placenta RNA revealed the same sequence in the placenta MnSOD as that from multiple mouse organs. The data presented will be useful for wild-type sequence verification and possible point mutation detection. PMID- 8406028 TI - Cloning and sequencing of a cDNA encoding rat brain mitogen-activated protein (MAP) kinase activator. AB - The complete cDNA sequence for mitogen-activated protein kinase activator from rat brain was cloned. It encodes a protein kinase of 393 amino acids with a calculated M(r) of 43,465. PMID- 8406029 TI - Cloning and characterization of human TNF alpha promoter region. AB - We report the sequence of a 1.2-kb human tumor necrosis factor alpha (TNF alpha) promoter region, which was cloned using PCR. The sequence has several variations from two previous reports and exhibits many potential DNA-binding sites specific to mammalian gene regulatory proteins inducible by lipopolysaccharides. PMID- 8406030 TI - Promoter resurrection by activators--a minireview. AB - Frequently, in nature, defective promoters can be resurrected by activator proteins in response to cellular demands. The activators bind to nearby DNA sites for action. Various protein-protein and DNA-protein contacts involving activators, RNA polymerase, and different segments of DNA in and around a defective promoter form a DNA-multiprotein complex (cage) which enhances transcription. PMID- 8406031 TI - Genes required for extracellular secretion of enterotoxin are clustered in Vibrio cholerae. AB - Pleiotropic transposon insertion mutants of Vibrio cholerae that are unable to secrete enterotoxin, HA/protease and chitinase through the outer membrane have been isolated. The gene, epsM, responsible for complementation of two of the Tn5 insertion mutations was sequenced. It encodes a putative cytoplasmic membrane protein of 18.5 kDa that exhibits similarity to proteins required for extracellular secretion of pullulanase, pectate lyase or elastase in other Gram bacteria. It is present on a 15-kb DNA fragment from the V. cholerae genome, containing the epsE gene that was previously shown to be required for secretion of cholera toxin [Sandkvist et al., Gene 123 (1993) 81-86]. Partial reading frames flanking epsM also demonstrated similarity to genes required for extracellular secretion of pullulanase in Klebsiella oxytoca. PMID- 8406032 TI - Complete sequence of omp1, the structural gene encoding the 40-kDa outer membrane protein of Fusobacterium nucleatum strain Fev1. AB - The sequence of the omp1 gene coding for the 40-kDa outer membrane protein (OMP) of the Gram- oral bacterium, Fusobacterium nucleatum strain Fev1, has been determined. Degenerate oligodeoxyribonucleotide primers were used to prime the amplification of a 120-mer sequence of the gene. This sequence was successively used for constructing new primers applied in asymmetrical, symmetrical, and inverse polymerase chain reaction using as template genomic DNA, self-ligated DNA fragments, or fragments ligated into either pGEM-7Zf+ or pACYC184. The codon usage of the gene was unusual in that A or T was used as the third base in the codon triplets in all cases, except for those amino acids (aa) which have only one or two possible codon choices. Only 35 of the 61 sense codons were used. The aa sequence of the protein was deduced; it consisted of 348 aa (M(r) 39,954), which is in good agreement with the 40-kDa size estimated from electrophoretic analyses. The mature protein was preceded by a 20-aa signal peptide. PMID- 8406033 TI - Isolation and expression in Escherichia coli of a Xanthomonas oryzae recA-like gene. AB - The recA gene from the bacterium Xanthomonas oryzae pv. oryzae (Xoo), a rice pathogen, was cloned based on its ability to complement DNA repair defects of Escherichia coli recA- mutants. The Xoo recA was localized to a 1.3-kb Sau3AI XhoI fragment and, when cloned into pBR322, specifies increased methylmethanesulfonate and mitomycin C resistance to E. coli recA mutants and allows lambda red- gam- to plaque on an E. coli recA- host. An E. coli recA- strain harboring a plasmid containing the Xoo recA-like gene was shown to produce a 40-kDa protein which cross-reacted with an anti-E. coli RecA antibody. A similar molecular mass protein to RecA has been detected in several Xanthomonas pathovars using an anti-E. coli RecA antibody. Furthermore, the cloned Xoo recA was shown to hybridize to genomic DNA from various Xanthomonas pathovars, but not to genomic DNA from other bacteria species under high-stringency hybridization conditions. These results indicate the isolation of the Xoo recA gene. PMID- 8406034 TI - MchAI and MchAII, two class-II restriction endonucleases from Mycobacterium chelonei. AB - We have purified and characterized two class-II restriction endonucleases from the saprophyte Mycobacterium chelonei AMB 82. MchAI was an isoschizomer of NotI recognizing and cleaving at 5'-GC/GGCCGC. MchAI did not require Triton X-100 for activity and was fully active at concentrations over 10-200 mM KCl and MgCl2. MchAII was an isoschizomer of HaeIII recognizing and cleaving at 5'-GG/CC. MchAII was active in 10 mM Tris.HCl pH 8.0 containing MgCl2 at 10-200 mM, was strongly inhibited by NaCl at concentrations higher than 10 mM and was inactive at KCl concentrations lower than 250 mM. PMID- 8406035 TI - Genetic manipulation of Campylobacter: evaluation of natural transformation and electro-transformation. AB - Two methods, natural transformation and electro-transformation, for the introduction of DNA into nine strains of Campylobacter jejuni were compared. Both methods were successful with a limited number of strains. Natural transformation was efficient only for the introduction of C. jejuni chromosomal DNA, while electro-transformation was also applicable for the introduction of Escherichia coli-derived vector DNA into at least one C. jejuni strain. The efficiency of DNA recombination after entry was determined using C. jejuni chromosomal DNA containing disrupted flagellin genes of C. jejuni or suicide vectors containing a portion of these genes. In the latter case, DNA recombination occurred with as little as 200-bp homology present, indicating that only short homologous DNA segments are required. PMID- 8406036 TI - Diversity in the Chlamydia trachomatis histone homologue Hc2. AB - Chlamydia trachomatis elementary bodies contain two developmentally expressed histone H1 homologues. An 18-kDa histone homologue, Hc1, is conserved among C. trachomatis serovars and C. psittaci. The other histone homologue, Hc2 (encoded by hctB), varies in size between C. trachomatis serovars but is present in reduced amounts or absent from C. psittaci. The variation in Hc2 size among C. trachomatis serovars was found to be due to internal deletions from a region of the hctB gene encoding lysine- and alanine-rich pentameric repeats. PMID- 8406037 TI - The glutamate dehydrogenase-encoding gene of the hyperthermophilic archaeon Pyrococcus furiosus: sequence, transcription and analysis of the deduced amino acid sequence. AB - Glutamate dehydrogenase (GDH) from the hyperthermophilic archaeon, Pyrococcus woesei, has been isolated, characterized and found to be very similar if not identical to the recently purified GDH from P. furiosus. Using a polymerase chain reaction, based on the N-terminal amino acid sequences of GDH, the P. furiosus gdh gene was identified, cloned into Escherichia coli and sequenced. The transcription start point of gdh has been mapped 1 nucleotide upstream from the ribosome-binding site. Using antiserum raised against purified GDH, expression of gdh was observed in E. coli. The deduced primary sequence of the P. furiosus GDH has been compared to various bacterial, archaeal and eukaryal GDHs and showed a high degree of similarity (32-52%). PMID- 8406038 TI - Diphthamide synthesis in Saccharomyces cerevisiae: structure of the DPH2 gene. AB - A gene involved in diphthamide biosynthesis, DPH2, was cloned from Saccharomyces cerevisiae by complementation of a diphthamide mutant. DPH2 exists as a single copy gene in the yeast genome and is located on the left arm of chromosome XI. Sequence analysis of the DPH2 locus predicts that the DPH2 gene product is a 534 amino acid (aa) protein, with a calculated M(r) of 59,772. This conclusion was supported by Northern blot analysis of the DPH2 transcript and gel analysis of the DPH2 protein overproduced in Escherichia coli. Gene disruption studies indicate that the DPH2 gene is not essential for viability of yeast. The role of DPH2 in diphthamide biosynthesis is discussed. PMID- 8406039 TI - Natural variation of the NgoII restriction-modification system of Neisseria gonorrhoeae. AB - The NgoII restriction-modification (R-M) system of Neisseria gonorrhoeae recognizes the sequence 5'-GGCC-3'. This system is encoded by two separate genes, dcmB for the methyltransferase (MTase) and dcrB for the restriction endonuclease (ENase). Three strains that vary in their NgoII phenotype were examined. Strain Pgh3-2 produced detectable levels of both enzymes, strain F62 lacked detectable levels of the dcrB gene product, and strain WR302 failed to produce either gene product. Strains that lacked either enzyme activity still possessed the genes that encode them. Transcriptional fusions of dcrB in strains F62 and Pgh3-2 indicate that this gene is transcribed at nearly identical levels in each strain. The DNA encoding the NgoII R-M system was cloned from the three strains, and the nucleotide sequence was determined. The dcrB genes of WR302 and F62 possess the same frameshift mutation (base position 1435) which would result in a truncated protein. The WR302 dcmB was found to have a point mutation that changed Arg288 (a residue that is conserved in all prokaryotic and phage cytosine MTases sequenced to date) to Trp. PMID- 8406040 TI - High-level heterologous gene expression in Saccharomyces cerevisiae from a stable 2 microns plasmid system. AB - The best candidate for a high-copy-number and mitotic stability expression system in yeast is the endogenous 2 microns plasmid. Nevertheless, derivatives of the 2 microns plasmid typically exhibit lower copy numbers and require selection for adequate maintenance within cells. We report the construction and utilization of an efficient heterologous gene expression system containing a 4.5-kb inducible expression cassette inserted into the 2 microns plasmid and selected in cells utilizing a carrier plasmid which is subsequently lost via FRT/Flp recombination. The non-selectable 2 micron plasmid, containing the cassette, was found to be stably maintained in cells, without selection, at high copy number. The dynamics of resolution and partitioning of this plasmid were analyzed during the course of 50 generations of growth under non-selective conditions. The heterologous lacZ reporter gene coding for beta-galactosidase (beta Gal) is driven by the hybrid, galactose-inducible promoter GAL10::pMF alpha 1. Upon induction, beta Gal was secreted into the periplasm and culture supernatant at levels which could be detected directly from Coomassie blue-stained SDS-PAGE. Furthermore, plasmid containing cells could be maintained directly on rich YPD medium and identified either by utilizing XGal or by observing inhibition of colony growth on YPGal solid medium. The cassette was designed for direct, high-level, inducible expression of cloned genes downstream from the MF alpha 1 signal sequence, with or without a C-terminal lacZ fusion. This vector represents the first demonstration of a non-selectable, mitotically stable, episomal plasmid system capable of expressing recombinant proteins at high levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406041 TI - Protein farnesyltransferase: production in Escherichia coli and immunoaffinity purification of the heterodimer from Saccharomyces cerevisiae. AB - Protein farnesylation in Saccharomyces cerevisiae is mediated by a heterodimeric enzyme, protein farnesyltransferase (PFTase), encoded by the genes RAM1 and RAM2. A series of plasmids for the expression of RAM1 and RAM2 in Escherichia coli was prepared and evaluated. Maximal production of functional PFTase was seen in strains containing a multicopy plasmid with a synthetic operon in which the RAM1 and RAM2 structural genes were translationally coupled by overlapping TAATG stop start codons and by locating a ribosome-binding site near the 3' end of the upstream gene. This was accomplished by an insertional mutation at the 3'-end of RAM1 that embedded an AGGAGGAG sequence within codons for the tetrapeptide, QEEF, added to the end of the Ram1 protein. The QEEF C-terminal motif in the Ram1 subunit of PFTase facilitated purification of the enzyme by immunoaffinity chromatography on an anti-alpha-tubulin column prepared using monoclonal antibodies that recognized a tripeptide EEF epitope. Heterodimeric recombinant yeast PFTase::QEEF (re-PFTase::QEEF) constituted approximately 4% of total soluble protein in induced cells and was readily purified 25-fold in two steps by ion exchange and immunoaffinity chromatography in an overall 25% yield. Michaelis constants for farnesyl diphosphate (FPP) and Hras protein (modified to contain a yeast a-mating factor PACVIA sequence at the C terminus) were 5.5 and 15 microM, respectively; the kcat was 0.7 s-1. PMID- 8406042 TI - A Saccharomyces cerevisiae upstream activating sequence mediates induction of peroxisome proliferation by fatty acids. AB - Peroxisome proliferation in Saccharomyces cerevisiae is induced by fatty acids via as yet unknown mechanisms. We have initiated a study of these mechanisms by identifying control sequences sufficient for fatty acid control of the CTA1 gene (encoding the peroxisomal catalase A). Promoter regions previously shown to be necessary for control were tested for their potential to mediate induction by oleic acid to a CYC1::lacZ fusion gene. A region previously demonstrated to control CTA1 via the ADR1 transcription activator (bp -156 to -184) does not mediate induction by oleic acid. In contrast, an adjacent sequence (-184 to -198) is sufficient for oleic acid induction, and a neighbouring element (-197 to -215) has marginal inducing activity. These two elements are characterized by a consensus sequence, 5'-CGGNNNTNA ('peroxisome box'), which is found in a number of S. cerevisiae peroxisomal protein-encoding genes. Mutation of either the CGG or the TNA block in the box has a dramatic down-regulating effect on the gene expression in oleic acid medium. PMID- 8406043 TI - Isolation and characterization of SSE1 and SSE2, new members of the yeast HSP70 multigene family. AB - Two new members of the Saccharomyces cerevisiae heat-shock protein 70 multigene (HSP70) family were isolated from a yeast expression library using antisera made against a yeast calmodulin-binding fraction. They are designated as SSE1 and SSE2, because their predicted amino acid (aa) sequences are highly homologous to each other (76% identical), and share homology with known members of the yeast HSP70 multigene family, but their homologies (13 to 28% identity) are not high enough to place them in known subfamilies. SSE1 and SSE2 are thought to encode polypeptides of 693 aa with calculated M(r)'s of 77,408 and 77,619, respectively. The SSE1 mRNAs were moderately abundant during steady-state growth at 23 degrees C, and increased a few-fold upon upshift to 37 degrees C. SSE2 mRNAs were present at low level during steady-state growth at 23 degrees C, and greatly increased upon upshift to 37 degrees C. Disruption of SSE1 results in slow-growing cells at any temperature. No phenotypic effects of the mutation in SSE2 were detected, and the growth property of the sse1sse2 double mutant was the same as that of the sse1 single mutant. PMID- 8406044 TI - Autoregulation of the gene encoding the replication terminator protein of Bacillus subtilis. AB - One of two putative sigma A promoters identified previously in the region immediately upstream from the rtp gene (encoding the replication terminator protein) [Smith and Wake, J. Bacteriol. 170 (1988) 4083-4090] has been shown by transcription start point (tsp) mapping to be the functional rtp promoter. In these tsp mapping experiments, it was observed that the level of mRNA from this promoter, Prtp, was increased by a factor of 30 in the absence of the replication terminator protein (RTP), consistent with the autoregulation of rtp at the level of transcription. In vitro transcription from Prtp by sigma A RNA polymerase has been shown to be specifically repressed by RTP. A Prtp-spoVG-lacZ fusion was inserted into the chromosome of a strain in which RTP production was inducible by IPTG. Addition of IPTG to cultures of the new strain lowered beta Gal production by a factor of at least four. It is concluded that rtp is autoregulated in vivo at the level of transcription. PMID- 8406045 TI - Isolation of a mitotic-like cyclin homologue from the protozoan Trypanosoma brucei. AB - Activation of the p34cdc2 protein kinase (PK) at different stages of the eukaryotic cell cycle is controlled by interaction with regulatory proteins known as cyclins (CYCs). Using a probe obtained by PCR amplification, we have isolated from the protozoan, Trypanosoma brucei, a cDNA clone encoding a CYC homologue. The amino acid sequence deduced for this gene (CYC1) shares structural homology with A- and B-type CYCs of other organisms, including a motif, the destruction box, which has been related to the rapid turnover of these CYC proteins in mitosis. When expressed in fission yeast, CYC1 is able to rescue the defect of a temperature-sensitive cdc13 mutant, demonstrating that it is functional as a cell cycle regulator. In trypanosome cells, CYC1 associates with a 34-kDa protein that cross-reacts with a monoclonal antibody against the conserved 'PSTAIR' epitope of p34cdc2, and the complex displays histone H1 PK activity. Furthermore, when trypanosome cells are synchronized by hydroxyurea treatment, CYC1 accumulates as cells progress towards mitosis. These observations, taken together, suggest that CYC1 is a component of the active PK complex required for the control of trypanosome mitosis. PMID- 8406046 TI - Improved bacterial hosts for regulated expression of genes from lambda pL plasmid vectors. AB - The construction and use of a set of Escherichia coli strains with defective lambda prophages that facilitate expression of genes cloned in lambda pL-plasmid vectors is described. These bacteria allow high and regulated expression of such genes, whereas a kanamycin-resistance marker (KmR) on the prophage allows easy identification and genetic transfer from strain to strain. Optimal conditions for examining gene expression with the pL-vector systems using these strains are discussed. PMID- 8406047 TI - Isolation of enterohemolysin (Ehly2)-associated sequences encoded on temperate phages of Escherichia coli. AB - We have cloned and sequenced the enterohemolysin (ehl)-associated region from a temperate bacteriophage isolated from an Escherichia coli O26:H11 strain. Phage phi C3208 was isolated together with other temperate bacteriophages which transduce the enterohemolytic phenotype to non-hemolytic E. coli O26 strains. The nucleotide sequence of the 1245-bp phi C3208 DNA insert in plasmid pEO39, which mediates Ehly2 production in E. coli K-12, was determined and was found to be partially homologous to DNA of bacteriophage lambda but is completely unrelated to DNA sequences encoding the synthesis of Ehly1 [Stroeher et al. Gene 132 (1993), 89-94]. It was shown that part of this region can be used as an Hly2 associated specific DNA probe. PMID- 8406048 TI - Tough business, this dying. Patients nearing death challenge physicians to provide appropriate care. PMID- 8406049 TI - Osteoporosis: patient workup shows not all cases are postmenopausal. PMID- 8406050 TI - Diabetes and heart disease: a new strategy for managing lipid disorders. AB - Dyslipidemias represent an underdiagnosed and undertreated clinical problem in the management of diabetic patients. Glycemic control by itself is not sufficient to correct elevated triglycerides and low HDL levels, which greatly increase the risk of cardiovascular disease. Careful monitoring and aggressive intervention can dramatically reduce the risk that these dyslipidemias pose in diabetic and prediabetic patients. Weight loss by obese patients, low-fat diets, and gradually increased aerobic exercise should be tried for 6 months. If lipid levels are still outside the acceptable range, consider adding lipid-lowering drug therapy. Age should be no barrier to intervention, as coronary risk factors continue to contribute to the incidence of events into advanced age. PMID- 8406051 TI - Attack of the scabies: what to do when an outbreak occurs. AB - Scabies remains a significant source of morbidity in nursing home residents because of its highly contagious nature. It is characterized by severe pruritus and papules, pustules, burrows, nodules, and occasionally urticarial lesions. Lesions are commonly found on the wrists, finger webs, antecubital fossae, axillae, areolae, periumbilical region, lower abdomen, genitals, and buttocks. Diagnosis is based on the history, physical examination, and demonstration of mites, eggs, or scybala on microscopic examination. Several topical scabicides are available, but permethrin cream appears to be less toxic and more effective in cases that are resistant to other agents. Successful management requires evaluation of individuals with close patient contact. PMID- 8406052 TI - How to make sure your older patients are getting enough zinc. AB - Older patients with inadequate dietary habits tend to be at risk for mild to moderate zinc deficiency. Symptoms that can be of particular concern include slow wound healing, increased risk of infection, and a loss of acuity in taste and smell. The diagnosis of zinc deficiency is based on a review of the patient's eating habits; laboratory testing is not generally useful in the clinical setting. Most older adults can achieve an adequate zinc intake by eating a variety of foods each day, including meat, fish, and poultry. Supplementation is appropriate in cases of known or suspected frank zinc deficiency. PMID- 8406053 TI - Aches and pains: tips on how to make their misery go away. PMID- 8406054 TI - Effect of age on the ability of rat liver tissues to transform chemical promutagens to mutagens. AB - Age-related changes in drug metabolism of the liver from male Lou rats were determined by measuring changes in the production of mutagens. Activation of aflatoxin B1 (AFB1), 2-aminoanthracene (2AA) and benzo[a]pyrene (B[a]P) to mutagenic derivatives was assayed using the Ames salmonella test system. The promutagens were incubated with tissue fractions isolated from rats ranging in age from 3 weeks to 18 months. Hepatic activation of AFB1 and 2AA changes with age and is maximal from 4 to 10 months. In contrast, no significant modification with age was observed for the activation of B[a]P. It is concluded that the composition of different cytochrome P-450 fractions is modified with age thereby altering the specific ability to convert different promutagens to mutagens. PMID- 8406055 TI - Ischemia-induced changes in brain monoamine metabolism in aged spontaneously hypertensive rats. AB - Effects of aging on monoamine metabolism in transient cerebral ischemia were studied using adult and aged female spontaneously hypertensive rats (SHRs). Tissue monoamine contents in discrete brain areas were quantified after 20 min of cerebral ischemia with or without 30-min recirculation. Cerebral blood flow (CBF) was determined with a hydrogen clearance method in separate experiments. Dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) contents in the striatum, nucleus accumbens and septum decreased in ischemic aged SHRs, compared with those in ischemic adult SHRs. After 30-min recirculation, DA contents were actually unchanged with inconsistently increased DOPAC levels. Ischemic CBF decreased to < 20% of the resting CBF in the striatum and cortex, which was not different between the two age groups of SHRs. These results indicate that aging is primarily responsible for more severely impaired DA metabolism during cerebral ischemia in aged SHRs. PMID- 8406056 TI - Elevated serum copper is associated with reduced immune response in aging mice. AB - Young mice were found to have serum copper concentrations ranging from a low of 0.291 to a high of 0.584 ppm. Old mice had serum copper concentrations ranging from 0.223 to 1.715 with 30.9% of the old animals having values greater than 0.6 ppm. The mitogen response of isolated lymphocytes from the spleens of aging mice was greatly reduced when these cells were taken from animals with naturally occurring serum copper levels in excess of 0.6 ng of copper/mg wet weight serum. The lymphocytes taken from young mice with higher serum copper concentrations, on the other hand, had increased response to mitogens. Addition of the copper protein, ceruloplasmin, to lymphocyte cultures in vitro reduced the mitogen response of purified splenic lymphocytes with the reduction being greater for cells from old animals. We suggest that excess serum copper and ceruloplasmin may be immunosuppressive, especially in older organisms. PMID- 8406057 TI - Changes in parietal and mucous cell mass in the gastric mucosa of normal subjects with age: a morphometric study. AB - Whether or not the gastric mucosa undergoes significant changes in normal aging subjects is still open to debate. In 51 subjects undergoing endoscopy and lacking any significant endoscopic or histologic modification we evaluated mucosal thickness, gland number, numbers of parietal, chief and mucous cells at the fundus and of mucopeptic cells at the antrum, with a morphometric method, subgrouping the patients according to their age class. Our findings demonstrate that the number of parietal cells tends to increase with age and, on the other hand, the number of mucous cells is reduced in elderly subjects (p < 0.05). When considering the parietal-to-mucous cell ratio, this is significantly increased (p = 0.0005) with age. Acid secretion being an offensive factor and mucus a fundamental component of the gastric mucosal barrier, these findings suggest an increased susceptibility of the gastric mucosa to damage in the elderly. PMID- 8406058 TI - Loss of calcium-homeostatic mechanisms in polymorphonuclear leukocytes of demented and nondemented elderly subjects. AB - The calcium homeostatic mechanisms were explored in 31 aged individuals some of whom suffered from dementia (11 Alzheimer's disease, 5 other dementias, 15 cognitively normal individuals). Their mean age was 81.2 +/- 9.2 years, varying from 60 to 96 years. Polymorphonuclear leukocytes (PMNs) were isolated from freshly drawn citrated blood and loaded with Fura-2. Resting intracellular free calcium values, [Ca2+]o, were recorded as well as the calcium transients produced by formyl-methionyl-leucyl-phenylalanine 10(-8) M:delta [Ca2+]i. The return to prestimulation values was also recorded. Average [Ca2+]o was 64.6 +/- 16.0 nM and the average delta [Ca2+]i 77.6 +/- 52.7 nM. No correlation was found with age within the range explored nor was there any difference in calcium transients between demented and nondemented patients. The most surprising finding was the inability of about 30% of all examined patients' PMNs to return the elevated [Ca2+]i values to prestimulation values. For some patients, elevated [Ca2+]i remained at the higher value, for others [Ca2+]i continued to rise. Those who could down-regulate poststimulation values did it only very slowly. Previous results obtained with leukocytes from younger individuals (under 45 years) showed a rapid return to prestimulation values. It appears that this loss of calcium homeostatic mechanisms in old individuals is a progressive process which involves a relatively high proportion of the investigated subjects. The rate of decrease of intracellular calcium (measured by the slope P) did not correlate with diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406059 TI - Guillain-Barre syndrome in the elderly. A retrospective comparative study. AB - There have been no reported studies specifically devoted to the Guillain-Barre syndrome in elderly patients. The survey reported in this paper investigated the manifestations of the disease in older patients and compared them with those seen in younger patients. All adult patients with an established diagnosis of Guillain Barre syndrome admitted over 9 years to a Regional Referral Centre providing services for a population of 543,213 were identified and studied. Fifteen patients were over the age of 60 years and 21 between 19 and 60 years of age. Antecedent illnesses and, in particular, gastro-intestinal symptoms were reported less frequently in older patients. Where there was an antecedent illness, this was of shorter duration in elderly subjects. Neurological symptoms related to peripheral nerve damage, such as paraesthesiae, myalgia, backache and an inability to stand, occurred with a similar frequency in both age groups. Double vision and facial weakness, however, occurred less often in older patients, and cerebellar features were not encountered in the older age group. The cerebrospinal fluid immunoglobulin abnormality was more marked in the elderly. The interval between onset and peak severity was shorter in elderly patients. The duration of hospital stay was significantly greater in the elderly, though the overall prognosis was similar in both age groups. There was only one death in each group. The incidence of the Guillain-Barre syndrome in the elderly was higher than in subjects aged 19-60 years, but this difference was not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406060 TI - The effects of nicotine on the blood flow of the ophthalmic artery and the finger circulation. AB - Cigarette smoking is associated with glaucoma. Nicotine from cigarette smoking is known to produce hemodynamic changes. We studied the effect of nicotine on the ophthalmic artery flow velocity using transcranial Doppler ultrasound to measure noninvasively the Peak (systolic) flow velocity (VP), mean-envelope flow velocity (VM) and end-diastolic flow velocity, as well as using a laser Doppler flowmeter on the finger blood flow of the finger prior to and after administration of 2-4 mg nicotine in a gum base. In a pilot project a group of 18 glaucoma patients and 8 normal subjects were tested with nicotine, and another group of 11 glaucoma patients were tested with placebo. VP, VM, and systolic blood pressure were significantly increased while finger blood flow was significantly decreased when comparing the nicotine-tested glaucoma group with the placebo-tested group (P = 0.02074, 0.01479, 0.02185, and 0.04209, respectively). The nicotine-tested group of normals also showed significant changes in VM, systolic blood pressure, and finger blood flow compared to a placebo group. The responses to the small doses of nicotine used in this study (approximately one-third that of cigarette smoking) were not significantly different between the glaucoma and the normal groups. PMID- 8406061 TI - The response of blood flow velocity in the ophthalmic artery and blood flow of the finger to warm and cold stimuli in glaucomatous patients. AB - Vasospasm appears to be a probable risk factor in the development of glaucoma. Transcranial Doppler (TCD) can detect vasospasm in cerebral vessels. It gives reproducible readings for ophthalmic artery blood velocity measurements. We used the TCD to measure peak flow velocity, mean flow velocity, and end-diastolic flow velocity in the ophthalmic artery and a laser Doppler flow meter to measure blood flow in the fingers of 17 patients with chronic open-angle glaucoma (COAG), 13 patients with normal tension glaucoma (NTG), and 8 nonglaucomatous subjects with normal eyes (normals). The opposite hand to that in which the measurements were made was challenged with warm (40 degrees) and cold (4 degrees) water. Five measurements of the ophthalmic artery velocity and finger flow were recorded: the first at baseline, the second during warming of the hand, the third after warming of the hand, the fourth when the hand was immersed in iced water, and the fifth after the hand was removed from the cold water. There was no detectable change in the ophthalmic artery blood velocities but significant decreases in the finger flow measurements when the opposite hand was immersed in cold water. These changes were present in all three diagnostic groups (P < 0.001 for COAG; P < 0.005 for NTG; P < 0.05 for normals). The findings suggest a reflex vasospasm to cold in the finger circulation of most persons but no measurable changes in the ophthalmic artery. PMID- 8406062 TI - Transscleral and indirect ophthalmoscope diode laser retinal photocoagulation: experimental quantification of the therapeutic range for their application in the treatment of retinopathy of prematurity. AB - Laser indirect ophthalmoscope (LIO) photocoagulation and transscleral photocoagulation through the conjunctiva and subconjunctiva were performed in the fundus of chinchilla gray rabbits using various exposure times and powers, and the thresholds for retinal blanching and choroidal hemorrhage were determined. The therapeutic range was described for both applications as the ratio between energy values to produce grayish white lesions and hemorrhage at 50% probability. The therapeutic range appeared to remain almost constant with different exposure times. The mean ratio with LIO was 3.2 +/- 0.28, similar to that with slit-lamp delivery reported in our previous study. The mean ratio with transscleral photocoagulation through the conjunctiva and subconjunctiva were 2.48 +/- 0.28 and 2.38 +/- 0.26, respectively. The variability of LIO appeared to be a little lower than with transscleral photocoagulation. There was no significant difference between the variability with transscleral photocoagulation through the conjunctiva and subconjunctiva. PMID- 8406063 TI - Two topical carbonic anhydrase inhibitors sezolamide and dorzolamide in Gelrite vehicle: a multiple-dose efficacy study. AB - The ocular hypotensive activities of the two potent topical carbonic anhydrase inhibitors sezolamide (previously known as MK-417) and dorzolamide (previously known as MK-507 and L-671,152) were compared formulated in Gelrite vehicle, a novel ophthalmic drug delivery system. This was a four-center, double-blind, randomized, placebo-controlled, parallel study in 73 patients with a diagnosis of bilateral primary open-angle glaucoma or ocular hypertension and a morning intraocular pressure (IOP) of greater than 23 mmHg in both eyes following washout of ocular hypotensive medications. Parallel 12-h modified diurnal curves were performed prestudy and on day 6, with a 4-h IOP curve on day 1. On day 6 the peak mean percentage decrease in IOP from baseline occurred 4 h after the dose of dorzolamide (22.1%) and 6 h after the dose of sezolamide (21.3%). There were no significant differences between 2% dorzolamide and 1.8% sezolamide at any time point, although the decrease in IOP for sezolamide tended to be slightly greater than that for dorzolamide. Duration of action of both compounds was, at most, slightly prolonged by the use of Gelrite vehicle when compared with former studies on sezolamide and dorzolamide. PMID- 8406064 TI - Late traction detachment in retinopathy of prematurity or ROP-like cases. AB - Five cases of traction retinal detachment occurring later in life as a sequel of cicatricial retinopathy of prematurity or showing the clinical picture of retinopathy of prematurity are reported. They presented with taut membranes in vitreous cavity, causing traction retinal detachment, and often showed preretinal membranes. These membranes were collagen-rich and contained cells with glial characteristics. They seemed to be continuously produced on the surface of the retina from which they detached sometimes in multiple generations. It is likely that chronic exudation from vascular abnormalities is a stimulus for this proliferation. These cases are very similar to other vitreoretinal proliferations in association with vascular abnormalities (Coats' and von Hippel disease, exudative vitreoretinopathy). PMID- 8406065 TI - Proton magnetic resonance spectroscopy of vitreal changes in experimental streptococcal endophthalmitis. AB - We used magnetic resonance spectroscopy to examine endophthalmitis in rabbits inoculated with a virulent strain of Streptococcus pneumoniae. On different days after infection, the animals were sacrificed and the vitreous isolated and examined with water-suppressed proton magnetic resonance spectroscopy. A broad resonance corresponding to the methyl envelope of lipoprotein lipids appeared 2 days after infection and persisted until the eyes developed phthisis (around 10 days postinfection). This resonance was absent in the control eye and the bacterial culture; it could be used as the marker of breakdown of blood-vitreous barrier and onset of endophthalmitis-induced changes. PMID- 8406066 TI - In vivo micropuncture of retinal vessels. AB - Micropuncture has proven to be a valuable tool for the local study of vascular parameters in many organ systems; however, it has not been applied to the study of the circulation of the retina. We report here our extension of micropuncture techniques [4] to use in the intact retina of the anesthetized cat. We use extremely sharp micropipettes with tip sizes much smaller than the diameter of erythrocytes to avoid hemorrhage. The micropipette is held by a microdrive which in turn is mounted on a precision goniometric micromanipulator. We micropuncture retinal arteries and veins with diameters ranging from 20 to 130 microns with no apparent damage to the vessel wall and no observed hemorrhage. During micropuncture we routinely inject nanoliter quantities of dyed saline, which we observe flowing in a plume from the micropipette tip within the lumen of the vessel. Micropuncture techniques may be used in the laboratory to study retinal autoregulatory mechanisms by microinfusion of vasoactive substances and by measuring blood pressure in retinal microvessels. In the clinic micropuncture may be useful for treating disorders such as retinal vascular occlusion. PMID- 8406067 TI - The effect of the ArF excimer laser on Candida albicans in vitro. AB - Candida albicans colonies in Sabouraud agar plates were irradiated with the excimer laser as follows: (a) at 10 Hz, power densities of 115-300 mJ/cm2 and 200 3000 pulses, and (b) power density of 115 mJ/cm2 and 200 pulses at 10-50 Hz. Additional colonies were irradiated with a power density of 200 mJ/cm2 at 10 Hz and 500 pulses. Each colony was cultured and the visible colony forming units were counted after 24 h. The cultures remained sterile at: 115 mJ/cm2, 1500 or 3000 pulses; 200 mJ/cm2, 400 or 500 pulses; and 300 mJ/cm2, 300 or 400 pulses. They decreased significantly in other groups. Photoablation with a power density of 115 mJ/cm2 and 200 pulses significantly decreased the number of yeast colonies in the culture plates at 30 Hz (p < 0.029) and 50 Hz (p < 0.02). Photoablation did not affect the counts in colonies located 1 or 2 mm from the treated colonies. Various energies of the excimer laser may significantly reduce or eliminate yeast in vitro. PMID- 8406068 TI - Inhibition of cytochrome oxidase and blue-light damage in rat retina. AB - The activity of cytochrome oxidase, outer nuclear layer thickness, and edema were quantitatively evaluated in the blue-light exposed rat retina. Dark-adapted or cyclic-light reared rats were exposed to blue light with a retinal dose of 380 kJ/m2. Immediately, 1, 2, and 3 day(s) after exposure, the retinas of six rats from each adaptation group were examined. There was no difference between the dark-adapted and cyclic-light reared rats. Immediately after light exposure, cytochrome oxidase activity decreased. The activity in the inner segments remained low at day 1, while severe edema was observed in the inner and outer segments. The outer nuclear layer thickness decreased 1-3 days after exposure. The blue-light exposure inhibited cytochrome oxidase activity and caused retinal injury. Similarity of the injury process in the dark-adapted and cyclic-light reared retinas suggests that rhodopsin was not involved. The inhibition of cytochrome oxidase could be a cause of retinal damage. PMID- 8406069 TI - Lens epithelial cell response to isoforms of platelet-derived growth factor. AB - It has been reported previously that platelet-derived growth factor (PDGF) may play an important role in the regulation of lens growth and differentiation. To evaluate PDGF-induced effects at the cellular level, we investigated the response of cultured bovine lens epithelial cells (BLEC) to PDGF-AB, -AA, and -BB isoforms at the cellular level. Stimulation of BLEC with PDGF isoforms showed no increase in cell proliferation under the culture conditions of this study. In contrast, measurement of cytosolic free calcium concentration ([Ca2+]i), which has been shown to be an important second messenger for controlling multiple cellular processes in the lens, revealed a dose-dependent rise in [Ca2+]i upon stimulation with PDGF-AB and -BB isoforms. PDGF-AA used in similar concentrations was not effective. Our data suggest that PDGF-AB and -BB may play a role in the regulation of cellular functions in BLEC via modulation of intracellular calcium homeostasis. PMID- 8406070 TI - Adjunctive hyperbaric oxygen therapy for actinomycotic lacrimal canaliculitis. PMID- 8406071 TI - [Air pollution related morbidity in Zaporozhe]. PMID- 8406072 TI - [Sanitary and microbiological control of ocean water in biological pollution]. AB - Problems of coastal waters sanitary and bacteriologic inspection are discussed. Lactose positive E. coli and Enterococcus indexes as key, and Pseudomonas aeruginosa index as a supplementary one are suggested. Epidemiologically safe levels of these indexes with regard to bathing are offered. PMID- 8406073 TI - [Experimental study of the toxicity of dry activated hydrolysed sludge]. PMID- 8406074 TI - [Hygienic standardization of some biological pollutants in the water]. AB - Paper is devoted to problem of sanitary protection of water reservoirs from pollution by bioinsecticides and biosynthesis biologic waste. Experimental data on influence of biological pollutants on organoleptic water properties, water sanitary and toxicologic status are presented. Standards for open water bodies are recommended: dendrobacillin--6 x 10(4) org/l, trichodermin--2 x 10(4) org/l, grain moth pollen--0.05 mg/l, fodder yeast--0.08 mg/l, turingin--0.048 mg/l. PMID- 8406075 TI - [Asbestos in drinking water (review)]. PMID- 8406076 TI - [Organic pollution of water and soil in the Khramtsovsky's state farm of the Sverdlovsk region]. AB - Sanitary and chemical examination of soil and water was carried out in State farm "Khramtsovskiy" (Beloyarsk region, Sverdlovsk province), where intoxication cases during outcropping were detected by chromatomass analysis of organic pollution. Local pesticide levels exceeding the MACs as a potential health hazard were noted. PMID- 8406077 TI - [Genotoxic effects of the complex interaction of industrial factors]. AB - Results of genoepidemiological study of reproduction function of workers in plastic goods production are presented. Incidence of spontaneous abortions (SA) in workers' families was analyzed. Frequency of early and late SA in women working in hazard production and wives of men working in harmful production is discussed. Differences in SA incidence in hazardous risk groups and controls were discovered. PMID- 8406078 TI - [Hygienic evaluation of working conditions of acrylonitrile fiber processing at textile factories]. AB - Main unfavourable factors of working environment in acrylonitrilic fibres processing are high concentrations of dust, high noise levels, considerable share of hard manual operations. Complex of unfavourable factors leads to reduction of muscular endurance to static effort, prolongation of motor reaction to light irritation, decrease of cellular and humoral immunity of working women. Suggested effective measures reduced dust content in working zone air. PMID- 8406079 TI - [Content analysis of gas emission forming in the foam of the initial composite for manufacturing polymeric concrete]. AB - Gas emission in foaming of initial composition for manufacture of polymeric concrete was investigated. Emitted gas contained (mg/kg): acetone 6790-7264, chlorobenzol 18.0-19.4, diethylene glycol 0.16, triethanolamine 0.05, 4,4' diphenylmethane diizocyanate 0.09, n-hydrocarbons C2-C4, ethylene- and acetaldehyde oxides--little amounts. Acetone was a priority substance of the isolates. PMID- 8406080 TI - [Effect of vitamin supplementation on the metabolism of iron, copper and manganese in athletes]. AB - The intake of vitamins C, P or vitamin complex (ascorutine, thiamine, riboflavine, pyridoxine, cyan cobalamin, folic acid) influenced direct and indirect indexes of iron, copper, and manganese metabolism increasing excretion of these biotics through intestinal tract and kidneys of sportsmen skiers. Addition of vitamin complex with trace elements and macroelements to the ration improved iron and copper balance in the body and functional status of athletes better than vitamins alone. PMID- 8406081 TI - [Hygienic evaluation of the nutritional value of children's food in a pre-school facility and principles of its correction in the area affected by Chernobyl nuclear plant accident]. PMID- 8406082 TI - [Evaluation of the difficulty level of the subjects taught in high schools and colleges]. PMID- 8406083 TI - [Effectiveness of comprehensive medico-prophylactic measures in pre-school facilities]. AB - The study of health state of children visiting preschool institutions of the town of Tashkent showed that 80% them belonged to 2nd and 3rd health groups. Breathing exercises, point treating massage, therapeutic electrophoresis, galvanization, d'arsonvalization and others measures reduced disease incidence in children by half. PMID- 8406084 TI - [Evaluation of average effective doses of internal irradiation outside the regions polluted by the Chernobyl nuclear plant accident]. AB - Results of strontium-90 and cesium-137 control in food products in Russia after the Cernobyl power plant accident in 1986-1988 are summed up. Annual intake of radionuclides with individual food products and with ration on the whole was calculated. Maximal year intake of Sr-90 and Cs-137 with ration were accordingly 1% and 0.7% of the threshold annual intake for category B. Assessment of the mean effective equivalent doze (EED) of internal irradiation for Russia in 1986-1988 years was made; maximal addition to annual EED of internal irradiation realized in 1986 made up 2% from the mean annual EED of natural background. PMID- 8406085 TI - [Comparative evaluation of cancerogenic risk of radiation and air pollution by coal ashes and benzo(a)pyrene]. AB - Experiments were made with 800 white outbred mice inhaling benzo [a] pyrene (BP) and volatile coal ash for a long time. Experiments showed that BP is 10-1000-fold more carcinogenic than volatile coal ash. BP inhalation at a sanitary standard level (0.1 microgram/100 m3 of air) corresponds to equivalent risk of total gamma emission about 2 Sv. Coal ash inhalation in concentration 0.05 mg/m3 corresponds to the same equivalent risk as for the dose 0.05 Sv. PMID- 8406086 TI - [Effect of decis combined with radiation on skin tropism]. AB - Combined influence of detsius, sodium nitrate, total x-irradiation, and radon inhalations on rat skin was investigated. Preliminary irradiation of rats in a dose 100 R and 4-hour radon inhalation in concentration 8.1 x 10(7) Bq/m3 did not amplify detsius and sodium nitrate effects in LD50 doses. Detsius intake in 10- and 50-fold doses 5 times a week induced focal alopecia in a week. A conclusion is made on the necessity of prohibiting detsius use in practical agriculture. PMID- 8406087 TI - [Automated system of radiation control]. PMID- 8406088 TI - [Toxic action of resin - regulator of sugar beet growth]. PMID- 8406089 TI - [Rationale for minimal requirements in household chemicals]. PMID- 8406090 TI - [Hygienic assessment of substance transformation in indoor air ozonization]. PMID- 8406091 TI - [Cytogenetic activity of 2,4-D dimethylamine salt and ammonium nitrate in isolated and combined exposure of plants in experiment]. PMID- 8406092 TI - [Effect of gamma and electronic emission sterilization on sanitary and chemical characteristics of polypropylene ]. PMID- 8406093 TI - [Ecologic and hygienic problems of medical waste utilization abroad (review)]. PMID- 8406094 TI - [Medical and social aspects of developing a healthy life style in the rural population]. PMID- 8406095 TI - [Criteria significance of the accumulation count in hygienic analysis of xenobiotics mixtures]. PMID- 8406096 TI - [Prediction of pollution spreading in the near-earth atmospheric layer]. PMID- 8406097 TI - [Gas chromatographic determination of methyl ether of chloro carbonic acid in atmospheric air]. PMID- 8406098 TI - [Identification of zones with various levels of industrial air pollution based on space photography data]. AB - A methodological approaches and use of aerospace information for atmospheric air purity control are described. Image brightness was used as a major parameter for distinction of zones with different contamination levels. An optic contrast level of tested town zones was found changing in accordance with index of atmospheric pollution determined by ground means. PMID- 8406099 TI - [Investigation of the oil extraction process from various types of soil]. PMID- 8406100 TI - [Determination of some maleimides using a kinetic method]. PMID- 8406101 TI - [Determination of riboflavin in reservoir water using a fluorescent method]. PMID- 8406102 TI - [A filter burning device]. PMID- 8406103 TI - [Contribution of environmental factors regarding ontogenetic processes]. PMID- 8406104 TI - [The use of biological testing in hygienic assessment of water quality]. PMID- 8406105 TI - [Development of sanitary codex of 1939]. PMID- 8406106 TI - [The functional ovarian reserve in assisted reproduction. A clinical dilemma]. AB - The response to supraphysiological ovarian stimulation in Assisted Reproduction (particularly FIV-TE and GIFT) may be very heterogeneous, despite the fact that selected patients have clinical and paraclinical characteristics very homogeneous. Highly significant differences, were found, in two groups of patients with similar clinical characteristics, who were stimulated with the same scheme. Group I (Adequate response = 22 patients) and Group II (Inadequate response = 13 patients), regarding to basal seric levels of FSH (P = 0.007) and to quality of follicular response (P = 0.000). These results suggest that quality of response to supraphysiological ovarian stimulation, may reflect a "Functional ovarian reserve" and that this can be predicted (partially) with basal seric levels of FSH; but, may be, the inadequate response, paradoxical to certain point, would not represent an ovarian failure or a resistant ovarian syndrome, incipient or transitory during the reproductive age? PMID- 8406107 TI - [Homage to Dr. Martin Gimenez Miranda]. PMID- 8406108 TI - [Ethics and research on human reproduction]. PMID- 8406109 TI - [Ligation of the hypogastric arteries. Analysis of 4000 cases]. AB - Experience of 10 years is presented on hypogastric artery ligation as a secondary resource for the hemorrhage control on a selected group of 400 patients; those who have had a previous multiple management to the procedure. The pathology type is shown, risk groups and the problems that conditioned the indication. Surgical technique is analyzed, its complications and secueles; with a follow up of 2 years to 63.5%. Depurated mortality was of 1%. It is concluded that the procedure efficacy depends on it's prompt realization, adequate indication and good technique. PMID- 8406110 TI - [Gestational disorders]. AB - We present the results of an interview of 375 pregnant women which explores their gestational risk diagnoses, the types and frequencies of complaints. Five groups of 75 patients each were formed from this random sample corresponding to 30, 32, 34, 36 and 38 weeks of pregnancy respectively. We too analyze the patients' opinions about their own morbidity, the repercussion on activities of daily life and self-medication. The interview included the actions taken on their own account to deal with complaints during pregnancy, as well as to establish the relationships between these actions and medical attention they received. A total of 1534 complaints were recorded from the entire group, the mean per patient were four with a range from one to six. The reported from the entire group, the mean per patient were four with a range from one to six. The reported complaints did not caused medical consultation and were totally different with the respective risk diagnoses. Our results show that patients almost always accept their symptoms as a normal part of pregnancy, even when arise from pathology. Almost 70% of the patients view their symptoms as normal despite the fact that they produce difficulties of daily life in 41%, and that they temporarily block these activities in 19%. Around 10% of the patients admit self-medication. Over 50 types of actions were self-initiated, 1371 for the entire group as a whole. Around 95% of the complaints considered abnormal received no medical attention. We presented a series of thoughts on the effectiveness of popular practices and the possibility of integrating some of them into medical practice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406111 TI - [Therapeutic efficacy of vaginal administration of bromocryptin in patients with hyperprolactinemia]. AB - Oral bromoergocriptine (BEC) is currently the treatment of choice in women with hyperprolactinemia secondary to a prolactinoma. However, undesirable side effects (of variable type and intensity) are frequently present in these women due to both local irritation and to a direct effect upon the central nervous system. The present work was undertaken as a pilot study to assess the therapeutic effectiveness of vaginally administered BEC and to corroborate if the side effects are less frequent and of minor intensity when compared to oral BEC. Initially, 16 women were included, but two of them did not accept to continue in the study; thus, a total of 14 women with hyperprolactinemia (> or = 40 ng/ml) were divided in two groups: Group A encompassed five women, aged 27 to 36 years old, two with normal menstrual cycles and three with oligomenorrhea; all had primary or secondary sterility during 3 to 12 years and galactorrhea from 6 months to 3 years; in only one patient a brain computerized axial tomography (CAT) was performed which showed the existence of a macroprolactinoma. All received oral BEC (2.5-5 mg/day, except one patient with 10 mg/day). Group B included nine women, aged 26 to 36 years old, four had normal menstrual cycles and five had oligomenorrhea; all had primary or secondary sterility during the last 2 to 7 years and eight out of nine, also had galactorrhea during 1 to 8 years; in four of them a CAT was performed showing a pituitary microadenoma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406112 TI - [Efficacy and safety of azithromycin in the treatment of female genital Chlamydia trachomatis infections]. AB - Genital tract infections by Chlamydia trachomatis associated to sterility and infertility problems as well as perinatal complications have become increasingly frequent. Azithromycin is a new macrolide with a lower activity spectrum than erythromycin and a longer half life as well as less secondary effects. The objective of the study was to evaluate the safety and efficiency of Azithromycin on genital tract infection by C. trachomatis. MATERIAL AND METHODOLOGY. A total of 30 nonpregnant women between the ages of 19 and 35 were studied; 70% had only one sexual partner. In order to insure the presence of C. trachomatis as unique pathogen, cervicovaginal sampling, clinical evaluation and gynecologic exploration were undertaken. One dose of 1 g orally of Azithromycin was administered evaluating microbiologic and clinical remission at days 7-10, 12-16 and 33-37 after treatment. RESULTS. Two patients abandoned the study; global criteria of the evaluation were good to excellent in 17 cases; moderate to sufficient in six and poor in five. None of the cases reported secondary reactions. Results showed that Azithromycin treatment of cervicitis by C. trachomatis is useful with the advantage of unique dose administration. PMID- 8406113 TI - [Effect of a combination of gestoden-ethylene estradiol on the endometrium]. AB - It has been long established that contraceptive steroid mixtures are highly effective due to the fact that they act simultaneously upon different levels of the pituitary-ovarian-genital tract axis. We hypothesized that the mixture of 30 mcg of ethynylestradiol plus 75 mcg of gestodene might be effective partly through the changes it induces in the histology and histochemistry of the endometrium. To test this hypothesis, 32 healthy patients willing to participate, aged 23 to 34 years, were treated with the above mentioned combination of steroids from day 1 through day 21 of the cycle for six consecutive cycles. They were previously proven to be fertile and ovulatory. In ten of them, selected at random, an endometrial biopsy was taken on the cycle previous to therapy and upon completion of the steroid treatment, for comparison. Tissues were processed for histologic evaluation with hematoxylin-eosin and with acridine orange for the histochemical demonstration of glycogen and ribonucleic acid under fluorescence microscopy. All pre-treatment endometria showed normal findings for ovulatory cycles, while the post-treatment tissues were typical of the steroid-treated endometria, i.e., they had a disrupted balance between glandular and stromal maturation, plus a diminished and abnormally distributed glycogen and ribonucleic acid. We conclude that the established working hypothesis was confirmed. PMID- 8406114 TI - [Human papillomavirus infection in women with and without abnormal cervical cytology]. AB - This study sought to define the prevalence rates of human papillomavirus (HPV) infection and cytologic abnormalities in 3,257 sexually active females 13 to 45 years of age, undergoing routine cervical cytologic screening in the outpatient clinic of an urban hospital. One hundred and fifty patients (4.6%) showed cytologic evidence of cervical human papillomavirus infection (abnormal Pap). We selected a control group (n = 150) with negative cervical cytologic smears. Cells collected were analysed for HPV-DNA by PCR amplification method with probes for HPV types 6.11, 16.18 and 33. The HPV-DNA was detected in 21/300 (7.0%). The prevalence of cervical HPV-DNA types among women with negative cytology was 5.3% (8/150) and 8.6% (13/150) among women with abnormal Pap. The risk of HPV infection seems to be related with age at first intercourse, younger age and number of sexual partners. We did not find relation with the use of oral contraceptives smoking and history of prior sexually transmitted disease. PMID- 8406115 TI - [An orthotopic pregnancy after 2 ectopic ones. A report of 2 cases and review of the literature]. AB - The prognosis as to reproduction of patients with two surgical procedures for the tubes for managing ectopic pregnancy, is very bad. Two cases of live intrauterine pregnancy, after two tubal pregnancies surgically managed, are presented. Salpingectomy and salpingostomy were carried out in one patient, and salpingostomy and salpingectomy in the other one. In world literature there are only 23 reported cases, since 1947. PMID- 8406116 TI - [Uterine contraction induced by carbachol is inhibited by melatonin]. AB - It has been suggested that the pineal gland has a specific role in the regulation of reproductive functions. Melatonin, secreted by pineal gland, is involved in the control of mammalian reproduction. Previous investigations have show that melatonin reduced the smooth muscle contraction. The purpose of the present study was to investigate the effect of melatonin on the uterine contraction provoked by carbachol. This effect was studied in isolated uterus were taken from Wistar rats pretreated with diethylstilboestrol. Here, we describe the effects of various concentrations of melatonin was found to inhibit the carbachol-induced uterine contraction. Our results show that CE50 of carbachol increment in present to 10( 9) and 10(-6) M/ml of melatonin. The degree of the inhibitory effect of melatonin to concentration of 10(-9) M/ml is most evident that 10(-6) M/ml. Thus, it is concluded that melatonin has an pharmacological inhibition effect on the contraction uterine provoked by carbachol, act as physiological antagonist. PMID- 8406117 TI - [Immunity, the trophoblast and decidua]. AB - It is as if the embryo has learnt to foil the maternal inflammatory/immune system. Acting through trophoblastic protein-1, the conceptus inhibits prostaglandin F-2a synthesis and thereby prolongs corpus luteum survival. In addition, that protein (an interferon variant), suppresses T cell proliferation. Other interferons (b and g), prostaglandin E-2 and progesterone, synthesized by trophoblast and decidua, act in concert to help the embryo to elude the aggression of the immune system. Failure of these mechanisms leads to reactivation of maternal killer T cells, mononuclear infiltration in the conceptus and its demise. PMID- 8406118 TI - [The treatment of cervical human papillomavirus (HPV) infection with trichloroacetic acid]. AB - Ever since the alterations caused by the HPV on the cervix were discovered, countless treatments have been employed but the ideal method still remains unknown. The objective of the experiment was to assess the efficiency of handling the cervical infection caused by the HPV, by means of trichloroacetic acid, 85% rate. During the period comprised between April 1989 and March 1990, 60 patients were subject to close analysis. The general diagnosis drawn cytology, colposcopy, and histopathology was cervical condyloma, lacking any collateral evidence of intraepithelial cervical neoplasia. The treatment consisted of 3 weekly applications of trichloroacetic acid at 85% rate directly upon the cervix. They were all assessed through cytology and colposcopy every 3 months, during a year's period. After the first 3 months, a 73.4% healing rate was observed, which decreased to 68% after 6 months and to a 65% after 9 months; this last healing rate remained unchanged after 12 months. Pregnant patients showed higher healing rates. We have drawn the conclusions that the trichloroacetic acid is an efficient agent to treat the referred injuries, and it offers as well the advantages of low costs, no secondary effects and an easy application and handling. PMID- 8406119 TI - [Antibiotic use in pregnancy. II]. AB - Tetracyclines are antimicrobial agents that must be avoided during pregnancy. Metronidazole has showed to be carcinogenic in animals like mouse, but in humans this effect has not been proved. In developing countries the tuberculosis has a high prevalence, so tuberculosis in pregnant women isn't a rare event, all of this patient must receive antituberculosis agents. Of the extensive group of antiviral agents only few has proved clinical efficiency, of this acyclovir can be used during pregnancy with caution. At his moment there aren't sufficient information about the utility of zidovudine in pregnant women. PMID- 8406120 TI - [The endocrine capacity of the mammalian blastocyst. II. Biosynthesis and estrogen metabolism]. AB - Among the studies realized in preimplanted embryos, draw attention to its endocrine capacity to synthesize steroid hormones: progesterone, progestins and several estrogens, which may influence locally the properties of the oviduct and endometrium creating and appropriate environment for its nutrition, migration and further development in order to let its implantation in the maternal uterus. Among the steroids secreted by the conceptus, estrogens are of special interest because of their potential importance in the biochemical events associated with the process of implantation. The purpose of this review is to contribute to the knowledge of the biosynthesis and metabolism of estrogens realized by the early embryo. PMID- 8406121 TI - [The intensity of fetal movements vs. amnioscopy and cardiotocography in assessing fetal well-being]. AB - In 1989 at the Hospital Civil of Guadalajara we make a study with 100 patients with a term pregnancy, initial labor and alive fetus. At all women we practiced interrogatory about subjective intensity of fetal movements, amnioscopy and cardiotocographic registry and we compare results with Apgar score of the newborns at minute and five minutes. The fetal movements were normal in 91% of which 95.7% had Apgar score > or = 7 at minute and 98.9% Apgar score > or = 8 at five minutes; clear amniotic fluid in 75% of which 94.6% had Apgar score > or = 7 at minute and 100% had Apgar > or = 8 at five minutes and 94% with normal cardiotocographic registry of which 95.7% had Apgar score > or = 7 at minute and 100% had Apgar score > or = 8 at five minutes. In the distress fetal prediction, the best method was the cardiotocography and the worse was the meconial amniotic fluid. We conclude that the subjective intensity of fetal movements are similar in the well-being fetal prediction to the amnioscopy and cardiotocography and the physicians must use it in the obstetric care. PMID- 8406122 TI - [Serum prolactin during oral metoclopramide in normal nulliparous women]. AB - As a first step in an extensive project planned to determine serum PRL levels in response to oral metoclopramide in women with a diverse gyneco-obstetric history, it was decided to study 51 clinically healthy nulliparous women, aged 15.8 to 48.2 years, with history of regular menses at least one year before the study (except the three postmenopausal women), with no regular drug ingestion during the last six months. Women were studied on days 18 to 22 of menstrual period, after a 30 minute rest on basal conditions (3 samples) at 60, 90, and 120 minutes after a single 10 mg. oral dose of metoclopramide. Duplicate PRL determinations were performed in all samples and progesterone(P) only in a pool of the three basal samples by radioimmunoanalysis. All women had serum P levels > or = 4.0 ng/ml. A significant linear positive correlation (r > or = 0.6795, p < 0.001) was observed between chronologic age (CA) and serum PRL levels, regardless the way they were expressed. Considering the individual responses it was decided to divide the group according to CA and it was observed that serum PRL levels- expressed in any form were always significantly greater in women aged > 25 years (Group 2) in contrast with women aged < or = 25 years (Group 1). Since differences were evident, percentiles 3, 50 and 97 for serum PRL levels were calculated during each test time for both groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406123 TI - Fe(2+)-induced lipid peroxidation kinetics in liposomes: the role of surface Fe2+ concentration in switching the reaction from acceleration to decay. AB - Kinetics of malonyldialdehyde (MDA) accumulation, Fe2+ oxidation, and chemiluminescence (CL) at different initial iron ([Fe2+]) and liposome ([L]) concentrations were measured in liposome suspension. Above certain critical Fe2+ concentrations ([Fe2+]*) the latent period (LP) of LPO development was observed. The method of [Fe2+]* estimation by the dependence of LP value (tau) on [Fe2+] was elaborated. The increase of [L] resulted in decrease of tau and increase of delta MDA as well as SF CL amplitude. [Fe2+]* value changed from 10 to 50 microM with change of [L] from 1 to 4 mg/ml, so that the ratio [Fe2+]*/[L] was kept constant. This may be explained under the assumption that the major part of Fe2+ is bound by the membranes. At concentrations of Fe2+ higher than the critical one, iron chelators (desferrioxamine, o-phenanthroline, and EDTA) and cations (Eu3+, Ca2+, and Fe3+) decreased tau without any essential influence on the CL "slow flash" amplitude (h). Apparently, the only result of iron complexones and cations on LPO is the decrease of Fe2+ ion concentration on the membrane surface. Thus, [Fe2+]* value and surface concentration of Fe2+ are the main parameters determining both kinetics and efficiency of Fe(2+)-induced LPO in membrane systems. PMID- 8406124 TI - Effects of the xanthine oxidase system on cardiac function in anaesthetised rats. AB - This investigation aimed to determine whether contractile dysfunction of the myocardium could be produced upon generation of free radicals in the anaesthetised rat. The enzyme xanthine oxidase, combined with its substrate purine and an iron source, was used to generate free radicals in the venous circulation. The suspended form of xanthine oxidase, with substrate, produced a transient, significant depression in the contractile indices dP dt-1 max and dP dt-1 P-1 and arterial blood pressure, 1146 +/- 87 mm Hg s-1, 9 +/- 1 s-1, and 18 +/- 1 mm Hg, respectively. This could not be attenuated by the enzymatic free radical scavengers superoxide dismutase and catalase. Furthermore, the suspended xanthine oxidase alone or its vehicle were able to produce a similar effect to that of the complete free-radical-generating system. The maximum soluble dose of the crystalline form of the enzyme when employed in the generating system had no effect upon administration despite its production of superoxide radicals in vitro. These results suggest that the haemodynamic effects of the free-radical generating system containing the suspended form of xanthine oxidase were due to the effects of its vehicle and that the free-radical-generating system containing the crystalline form of the enzyme did not produce sufficient free radicals in vivo to modify myocardial contractility. PMID- 8406125 TI - Chemiluminescence from activated heme compounds detected in the reaction of various xenobiotics with oxyhemoglobin: comparison with several heme/hydrogen peroxide systems. AB - Chemiluminescence was detected in the reaction of oxyhemoglobin with various hydroxylamines and phenols, which have previously been shown to produce free radicals. The emitted light intensity correlated roughly with the methemoglobin formation rate, indicating the involvement of a photoemissive species as a reaction intermediate. In our previous work, we postulated the involvement of a catalase-insensitive, heme-bound hydrogen peroxide species in the methemoglobin formation reaction. In a series of experiments, we showed that intensive chemiluminescence occurred when hydrogen peroxide was mixed with either methemoglobin or metmyoglobin but not with hematin, which lacks the globin moiety. This suggests the involvement of the globin moiety in the light-emitting reaction sequence. The detection of paramagnetic globin species exhibiting similar kinetics as the corresponding light-emitting compound demonstrated that the assumed H2O2-heme compound has strong oxidizing properties. Accordingly, addition of bovine serum albumin to the hematin-hydrogen peroxide system also resulted in a strong chemiluminescence due to the formation of a paramagnetic transient species which could be detected by electron spin resonance (ESR). Several other heme compounds, such as cytochrome c or cytochrome c oxidase which have no vacant ligand site, did not show any light emission under similar conditions. This means that hydrogen peroxide must have access to a free-binding position on the heme. Chemiluminescence most probably stems from the transition of the initially formed heme-H2O2 adduct to the compound II type species. Due to their oxidizing nature, these species might be responsible for deleterious toxic effects such as lipid peroxidation and protein degradation. PMID- 8406126 TI - Oxyradical-mediated chromosome damage in patients with familial Mediterranean fever. AB - Increased chromosome breakage is observed in patients with familial mediterranean fever (FMF). Their plasma contains clastogenic material inducing chromosome damage in cells from healthy persons. It is proposed that increased oxyradical generation by activated polymorphonuclear cells in blood and serosal fluids of these patients leads to the formation of a clastogenic factor (CF), as it is observed in other chronic inflammatory diseases. Also similar to these diseases, the clastogenic effects are prevented by superoxide dismutase and partially by inhibitors of arachidonic acid metabolism. PMID- 8406127 TI - Oxidation resistance, oxidation rate, and extent of oxidation of human low density lipoprotein depend on the ratio of oleic acid content to linoleic acid content: studies in vitamin E deficient subjects. AB - The purpose of this study was to understand better the factors providing oxidation resistance to human low-density lipoprotein (LDL). Therefore, the susceptibility to copper-induced in vitro oxidation of LDL from vitamin E deficient patients and normal healthy subjects was studied. Surprisingly, the LDL of vitamin E deficient patients appeared less susceptible to oxidation than control LDL. Both oxidation rate and extent of oxidation, measured as diene production, were reduced when compared to control LDL. The lag time, a measure of resistance to oxidation, was not different from the lag time of LDL from healthy subjects. No relation was found between vitamin E content and resistance against oxidation. LDL from vitamin E deficient patients contained lower amounts of vitamin E, less cholesteryl esters, and increased amounts of triglycerides. Furthermore, its oleic acid content was increased and its linoleic acid content decreased. Linear regression analyses revealed that the ratio of oleic acid content to linoleic acid content was strongly correlated with the lag time, and inversely correlated with oxidation rate and extent of oxidation. Thus, LDL rich in oleic acid and poor in linoleic acid was less easily oxidized. It is concluded that the susceptibility of LDL to oxidation is determined not only by its antioxidant content, but also by other compositional factors, and more specifically by the ratio of oleic acid content to linoleic acid content. PMID- 8406128 TI - Measurement of n-alkanals and hydroxyalkenals in biological samples. AB - A modified method was developed to measure nM levels of a range of n-alkanals and hydroxyalkenals in biological samples such as blood plasma and tissue homogenates and also in Folch lipid extracts of these samples. Butylated hydroxytoluene (BHT) and desferrioxamine (Desferal) were added to samples to prevent artifactual peroxidation. Aldehydes were reacted with 1,3-cyclohexanedione (CHD), cleaned up by solid-phase extraction on a Sep-Pak C18 cartridge and the fluorescent decahydroacridine derivatives resolved by reverse-phase high-performance liquid chromatography (HPLC) with gradient elution. A wider range of aldehydes was detected in lipid extracts of plasma and liver homogenate compared to whole (unextracted) samples. Human plasma contained nM levels of acetaldehyde, propanal, butanal, pentanal, hexanal, and heptanal. 4-Hydroxynonenal (0.93 nmol/g) and alkanals with two to six carbons (up to 7.36 nmol/g) were detected in rat liver. Recovery of aldehydes added to whole plasma or to lipid extracts of plasma was dependent on carbon chain length, varying from 95% for acetaldehyde to 8% for decanal. Recovery from biological samples was significantly less than that of standards taken through the Sep-Pak clean-up procedure, suggesting that aldehydes can bind to plasma protein and lipid components. PMID- 8406129 TI - Effect of long-term exposure to cold on the antioxidant defense system in the rat. AB - Catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), and glutathione-S-transferase (GST) activities as well as glutathione (GSH), ascorbic acid (AsA), and vitamin E concentrations were analyzed in the blood, liver, brain, interscapular brown adipose tissue (IBAT), and small intestine of rats exposed to low environmental temperature (4 degrees C; 35, 75, and 105 d of exposure) and in controls of the same age exposed to an environmental temperature of 22 +/- 2 degrees C. Prolonged cold exposure resulted in an increase in GSH-Px in IBAT and in small intestine after 35, 75, and 105 d of exposure. Catalase activity in cold-exposed animals was higher in IBAT after 75 and 105 d of cold exposure. Glutathione reductase activity was greater in brain after 35 d, in liver after 75 d, and in IBAT after 105 d of exposure to low temperatures as compared to the controls. In contrast, GST activity was lower in liver and IBAT after 35 and 75 d of cold exposure. AsA and GSH (determined only 105 d after cold exposure) were markedly higher in IBAT, whereas plasma GSH was lower and plasma AsA was higher in cold-exposed animals. The observed changes in analysed components of the antioxidant defense system under conditions of prolonged exposure to low temperature suggest that a reorganization the activity of this system at the molecular level occurred. Although other studies indicate that a 21 d cold exposure is sufficient for adaptation of thermogenesis, the present study shows that in general, longer periods are required for the registration of the changes in the antioxidant defense system. PMID- 8406130 TI - Two pathways of iron-catalyzed oxidation of bilirubin: effect of desferrioxamine and trolox, and comparison with microsomal oxidation. AB - The bilirubin-degrading activity of liver microsomes from rats induced with 3 methylcholanthrene has been shown to be markedly stimulated by addition of 3,3',4,4',5,5'-hexabromobiphenyl, a polyhalogenated chemical which resembles in size and shape the most effective inducers of cytochrome P450IA1, but lacks the structural features necessary for it to be metabolised. The degradation of bilirubin by this microsomal system has been compared to oxidation by a chemical model system involving H2O2 and Fe-EDTA (ethylenediaminetetraacetic acid). In both systems bilirubin disappearance was accompanied by bleaching. However, when either desferrioxamine or Trolox were present in the chemical model system, the rate of bilirubin oxidation was greatly enhanced and, at the same time, bilirubin was largely or entirely converted to biliverdin, a pathway of oxidation which proceeds by dehydrogenation. In the presence of desferrioxamine, biliverdin was also further oxidised to an unidentified red pigment. PMID- 8406131 TI - Molecular pharmacology of vitamin E: structural aspects of antioxidant activity. AB - In this review, the involvement of vitamin E in free radical physiology and antioxidant mechanisms is discussed. Moreover, structure-activity relationship (SAR) studies on vitamin E analogues are presented. A molecular explanation for the antioxidant activity often is based on molecular parameters, such as Hammett sigma and Brown sigma +. These parameters correlate with the activity. Using semiempirical calculations, we have found other molecular parameters related to electron distribution and structure (such as the difference in heat of formation between the compound and its radical or the energy of the highest occupied molecular orbital, HOMO) which correlate with the antioxidant action of vitamin E and its derivatives. PMID- 8406132 TI - Alkylation and cleavage of DNA by carbon-centered radical metabolites. AB - Although carbon-centered radicals are formed during the metabolism of several genotoxic compounds, they have received little attention as DNA damaging agents. Carbon-centered radicals, however, can both cleave the DNA backbone and alkylate DNA bases, as has been demonstrated to occur in chemical and biochemical systems. Also, in vivo DNA alkylation by methyl radicals has been evidenced by isolation of C8-methylguanine in hydrolysates of DNA from rats administered 1,2 dimethylhydrazine. While most of the studies related to DNA damage by free radicals have been focused on oxyradicals, further studies on DNA alterations promoted by carbon-centered radicals may be necessary to elucidate the mechanisms of action of chemical mutagens and carcinogens. PMID- 8406133 TI - Spin trapping of nitrogen dioxide radical from photolytic decomposition of nitramines. AB - The photochemical (lambda < 400 nm) decomposition of some monocyclic and polycyclic nitramines produces .NO2, which can be detected in the respective nitramine crystals at 77 K by EPR (electron paramagnetic resonance). In solutions of perdeutero-dimethylsulfoxide (DMSO-d6) the .NO2 produced by photolytic decomposition of dissolved nitramines can be spintrapped by the solvent to give a radical having the structure CD3-(SO2)-(NO.)-CD3. In this article, we examine this reaction for two nitramines: cyclotrimethylenetrinitramine (RDX) and hexanitrohexaazaisowurzitane (HNIW), which are energetic materials. The decay of the spin-adduct radical (I) follows first-order kinetics for both nitramines studied, having a rate constant (k) of congruent to 7.1 x 10(-4) s-1. The net growth in spin concentration of (1) measured from EPR spectra is fitted by a first-order rate equation taking into account the simultaneous competitive decay rate of spin adduct (I). Using the rate data and EPR spin concentration data, the ratio of free .NO2 produced per parent nitramine molecule is estimated as 1:1 for RDX and 4:1 for HNIW. Biological implications of trapping of .NO2 by dimethyl sulfoxide are discussed. PMID- 8406134 TI - Evidence for free radical formation during human kidney transplantation. AB - Fourteen patients undergoing kidney transplantation were studied for evidence of the production of free radicals as assessed by the measurement of vitamin E (an index of lipid peroxidation) and of myeloperoxidase (a marker of neutrophil activation) in the systemic blood. Early (2 min) and late revascularization (30 min) of the kidney were respectively associated with a significant decrease of 35.5 and 40% of the initial level of plasma vitamin E. This consumption paralleled to the decrease of the vitamin E/total lipids ratio, a better indicator of vitamin E status. Heparin administration preceding renal artery clamping resulted in a twofold significant increase of baseline plasma myeloperoxidase (MPO) level (523 +/- 214 ng/ml). At kidney reperfusion, MPO concentration rose again and reached a maximum value of 1,653 +/- 882 ng/ml, indicating the presence of considerable neutrophil activation. A return to the baseline value was observed after 30 min of reperfusion. A short discussion about the possible origin of this MPO increase is given. Taken together, these data strongly suggest that free radical production, leading to lipid peroxidation phenomena, can occur within the early phase of kidney revascularization. Preliminary data using electron spin resonance with the spin-trapping technique strengthen this hypothesis. PMID- 8406135 TI - Memory and aging: components and processes. AB - This article provides a critical review of memory and aging. The focus is on the more accepted ternary scheme of memory, i.e. procedural, semantic and episodic, and on processing resources. A review of the literature in these areas, considering the more relevant studies or those with a greater number of subjects, reveals a gradual decrease of performance with age. No single hypothesis, either psychological or physiological, seems to be capable of explaining this decline. However, the hypothesis of a cognitive slowing during aging has an appealing simplicity and offers the chance to integrate the myriad of task-specific explanations that have proliferated in the literature. PMID- 8406136 TI - Dihydroergocryptine protects from acute experimental allergic encephalomyelitis in the rat. AB - The effect of dihydroergocryptine, a natural alkaloid derivative which exhibits D2 dopaminomimetic properties, has been studied in Lewis female rats with experimentally induced allergic encephalomyelitis. A chronic treatment with dihydroergocryptine started two days before immunization, induced a dramatic reduction of prolactin levels accompanied by a marked amelioration of neurological signs. In addition, the proliferative activity of splenic lymphocytes induced by the mitogen Concanavalin-A (Con-A) was reduced in dihydroergocryptine-treated animals. It is suggested that this effect is related to the ability of dihydroergocryptine to lower prolactin concentrations or also, partially, to a neuroprotective action of this drug. PMID- 8406137 TI - Fatty acids as markers of the blood-cerebrospinal fluid barrier function in man. AB - The blood-cerebrospinal fluid barrier (BCB) function in man is derived from the CSF:serum ratio of albumin (QAlb). In order to study other than BCB protein transport we compared CSF:serum ratios of fatty acids in patients revealing normal and increased QAlb values. Ratios of unsaturated fatty acids, which are involved in inflammation, increased significantly in patients with increased QAlb (p < 0.005). In contrast, CSF:serum ratios of saturated fatty acids increased only slightly. We conclude that characterisation of BCB lipid properties may be of particular interest regarding drug penetration into the CSF and lipid metabolism in inflammation. PMID- 8406138 TI - Cervical radiculopathy following botulinum toxin therapy for cervical dystonia. AB - The development of transient right-sided lower cervical root impairment is reported in a 49-year-old woman affected by cervical dystonia, who had previously received botulinum toxin injections into the left sternocleidomastoid and the right trapezius. The possibility of a causal relationship was discussed with consideration of: 1) the absence of intercurrent illness or trauma; 2) the positive correlation between sensory-motor disturbances and the botulinum toxin induced remission of cervical dystonia; 3) the reversibility of cervical radiculopathy (there was no recurrence in the two years following the interruption of the treatment). Magnetic resonance imaging investigation disclosed an abnormal cervical spine geometry together with moderate spondylosis. We feel that mechanical postural changes following the successful botulinum toxin treatment for cervical dystonia might have played some role in the development of cervical radiculopathy. Thus, skeletal abnormalities should be checked when attempting to correct muscle overactivity by botulinum toxin. PMID- 8406139 TI - Morphine inhibits the thalamic component of the subcortical somatosensory evoked potentials in rats. AB - Somatosensory evoked potentials (SEPs) were obtained by electrical stimulation of the volar surface of the rat forepaw. Subcortical evoked potentials were recorded from the skull overlying the contralateral somatosensory area of the cortex. Additionally, spinal evoked potentials were simultaneously recorded from the dura overlying the dorsal surface of the spinal cord at C3 level. The subcortical SEPs consisted of three peaks I, II, and III which arise from the spinal cord, the brain stem and the thalamus, respectively. The spinal evoked potentials consisted of a negative-positive wave. The latency of the negative peak of the spinal evoked potentials matched the peak latency of component I of the subcortical SEPs. Six doses of intravenous (i.v.) morphine were used: 0.5, 1.0, 2.5, 5.0, 10.0 and 20.0 mg/kg. The spinal evoked potentials and components I and II of the subcortical SEPs were not significantly affected by morphine whereas the amplitude of component III was inhibited (up to 50%) in a dose dependent manner. Optimal effect was achieved with the dose of 2.5 mg/kg. Microinjection of the optimal i.v. dose of morphine (25 micrograms/kg) into the thalamus showed a similar effect on wave III of subcortical SEPs. The effect of morphine was completely reversed by i.v. naloxone (0.25 mg/kg). In conclusion, these results indicate that the primary locus of morphine action on the subcortical SEPs could be at the thalamic level. PMID- 8406140 TI - Stimulatory effects on lactotrophs and crop-sac of interleukin-1 and interleukin 2 in pigeons (Columba livia). AB - The ultrastructural effects of human-recombinant interleukin-1 beta (IL-1 beta) and human-recombinant interleukin-2 (IL-2) on the crop-sac (the target organ for prolactin secretion in birds) and the anterior pituitary lactotrophs, were studied in pigeons (Columba livia). The intraventricular microinfusion of the two interleukins produced maximal crop-sac stimulation with milk-like secretion, as demonstrated by the observation of ultrastructural changes in the lactiferous areas through scanning and transmission electron microscopy of crop-sac mucosa. A marked activation of the anterior pituitary lactotrophs was also observed. Crop sac and pituitary lactotrophs stimulatory effects were prevented by a previous intraventricular treatment with monoclonal antibodies for IL-1 beta and IL-2 receptors, but not by an intraperitoneal administration of naloxone. The present results show that interleukins possess in pigeons marked stimulatory effects on prolactin secretion and that these are mediated by specific receptors. PMID- 8406141 TI - beta-Amyloid neurotoxicity. AB - beta-amyloid (beta A, 39-43 amino acids) deposition in brain parenchyma and vessel walls is a major pathological feature of Alzheimer's disease (AD). This is associated with degenerative changes of neuronal cell bodies and processes, and neuronal death. beta A, a portion of a larger transmembrane glycoprotein, has been reported to be toxic in several tissue culture models. The neurotoxic activity of beta A synthetic peptides is associated with their fibrillogenic capacity. However, in vivo studies on beta A neurotoxic activity have not proved conclusive and further investigations are necessary to establish the pathogenetic role of beta A in AD. PMID- 8406142 TI - The British Society of Gastroenterology. Annual meeting. 15-17 September 1993. Abstracts. PMID- 8406143 TI - Transplantation of cultured small bowel enterocytes. PMID- 8406144 TI - Treatment of hepatic metastases by cryotherapy and regional cytotoxic perfusion. PMID- 8406145 TI - Audit of the role of oesophageal manometry in clinical practice. AB - This oesophageal laboratory serves a population of 1.5 million. The study aimed to review referral patterns and assess the cost effectiveness of oesophageal manometry in clinical practice. All 276 consecutive manometry studies performed between 1988 and 1991 were reviewed. Reasons for referral in the 268 first referrals were: dysphagia 50.4%, non-cardiac chest pain 23.1%, gastro-oesophageal reflux disease 14.2%, connective tissue disease 11.2%, and 'other' 1.1%. Manometry was normal in 49.3%, showed achalasia in 17.9%, diffuse oesophageal spasm in 13.4%, connective tissue disease in 7.8%, hypertensive lower oesophageal sphincter in 4.5%, nutcracker oesophagus in 2.6%, and 'other' in 4.5%. A positive diagnosis was significantly more common if dysphagia was the reason for referral (65.9% v 35.3%, p < 0.01). A positive diagnosis was established in 60% of patients referred with connective tissue disease, 30.6% with non-cardiac chest pain, and 21.1% with gastro-oesophageal reflux disease. A positive diagnosis was significantly more common in connective tissue disease when symptoms were present (85% v 10%, p < 0.05). Management was changed in 48.9% of all patients because of manometry findings. The cost of each oesophageal manometry study was calculated to be 63.00 pounds: every change in patient management cost 129.00 pounds. In conclusion, oesophageal manometry changed management in over 20% of patients with non-cardiac chest pain or gastro-oesophageal reflux disease and in over 60% of those with dysphagia. It is, therefore, a useful and cost effective test in patients with these symptoms. PMID- 8406146 TI - Duodenal histology, ulceration, and Helicobacter pylori in the presence or absence of non-steroidal anti-inflammatory drugs. AB - Duodenitis and gastric metaplasia, which is often colonised by Helicobacter pylori (H pylori), are increasingly recognised for their importance in the pathogenesis of duodenal ulcers. The situation is not clear in patients receiving non-steroidal anti-inflammatory drugs (NSAIDs), who have a higher risk of peptic ulceration. The aim of this study was to identify the duodenal histological abnormalities in the presence or absence of NSAIDs, H pylori, and duodenal ulceration. Endoscopic duodenal biopsy specimens were taken from healthy looking mucosa of 172 patients (74 took NSAIDs, and 98 did not). Duodenitis was graded according to the degree of neutrophilic and plasma cell infiltration, villus height, Brunner's gland prolapse, and gastric metaplasia. The activity of duodenitis was dependent on the neutrophilic infiltration. A global score covering all the above factors was constructed, and H pylori in both the stomach and duodenum, was also assessed. The results showed that duodenitis with varying degrees of neutrophilic infiltration and gastric metaplasia was found in 20 patients (27%) taking NSAIDs, compared with 56 patients (57%) not taking NSAIDs (chi 2 = 16.24, p < 0.001). This degree of duodenitis was also found in 20 of 25 patients (80%) with duodenal ulcers, regardless of NSAID intake (chi 2 = 15.38, p < 0.001). Gastric metaplasia was identified in 20 patients (27%) receiving NSAIDs and 38 (39%) not receiving NSAIDs. Duodenal H pylori was only seen in patients with gastric metaplasia 10 (50%) receiving NSAIDs, and 34 (89%) not receiving NSAIDs. H pylori positive gastritis, and the combination of active duodenitis and gastric metaplasia were independent predictors of duodenal ulceration. It is concluded that active duodenitis is less common in patients taking NSAIDs but is strongly associated with gastric metaplasia, H pylori positive gastritis, and duodenal ulceration. These findings are relevant to the pathogenesis and treatment of duodenal ulcers in patients taking NSAIDs. PMID- 8406147 TI - Amoxicillin plus omeprazole versus triple therapy for eradication of Helicobacter pylori in duodenal ulcer disease: a prospective, randomized, and controlled study. AB - Treatment with amoxicillin and omeprazole resulted in encouraging Helicobacter pylori eradication rates in pilot studies that included medium term follow up. These results were evaluated in a prospective, randomised and controlled study. Forty patients with active duodenal ulcer disease and H pylori colonisation of the gastric mucosa were randomly assigned to receive either omeprazole (20 mg twice daily) and amoxicillin suspension (500 mg four times daily) for two weeks (group I) or bismuth subsalicylate (600 mg three times daily), metronidazole (400 mg three times daily), tetracycline (500 mg three times daily), and ranitidine (300 mg in the evening) for two weeks (group II). Study medication was followed in both groups by a four week treatment course with 300 mg ranitidine up to the final examination. One patient from each group was lost to follow up. H pylori was eradicated in 78.9% of group I and 84.2% of group II (p = 1.00). All ulcers in patients on omeprazole plus amoxicillin healed but in the triple treatment group four patients had residual peptic lesions after six weeks (ulcer healing rate: 78.9%, p = 0.11). Complete pain relief occurred after a median duration of 1 day in group I and of 6 days in group II (p = 0.03). There were no major complications in either group but minor side effects were more frequently recorded in patients on triple therapy (63.2% v 15.8%, p < 0.01). In conclusion, two weeks of treatment with omeprazole plus amoxicillin is as good as triple therapy plus ranitidine in eradicating H pylori but seems better with regard to safety, pain relief, and ulcer healing. Thus, amoxicillin plus omeprazole should be recommended as the treatment of choice in eradicating H pylori in patients with duodenal ulcer disease. PMID- 8406148 TI - Disturbed gastric emptying in the short bowel syndrome. Evidence for a 'colonic brake'. AB - Gastric emptying of liquid (orange juice containing technetium-99m (99mTc) labelled antimony sulphide colloid) and solid (570 kcal pancake containing 0.5 mm resin microspheres labelled with Indium-111 (111-In)) was measured in seven patients with jejunum and no colon (jejunal lengths 30-160 cm), six patients with jejunum in continuity with the colon (jejunal length 25-75 cm), and in 12 normal subjects. In patients with no colon early emptying of liquid was rapid (median 25% emptying: 7 v 25 min, no colon v normal, p < 0.05); early gastric emptying of solid was rapid in two (each with less than 100 cm jejunum) and normal in the other five. Gastric emptying of liquid and solid for patients with jejunum in continuity with the colon was normal for the first three hours. There was increased liquid and solid retained in the stomach at six hours in both groups of patients (p < 0.01). Small bowel transit time was faster than in normal subjects for liquid in both groups of patients (p < 0.05) and for solid in those with no colon (p < 0.05). Rapid gastric emptying of liquid may contribute to the large stomal output in patients with a high jejunostomy. Preservation of the colon after a major small intestinal resection exerts a braking effect on the rate of early gastric emptying of liquid. PMID- 8406149 TI - The effect of liquid fibre on gastric emptying in the rat and humans and the distribution of small intestinal contents in the rat. AB - A combination of the polysaccharide ethyl-hydroxyethyl-cellulose (EHEC) and the surfactant sodium-dodecylsulphate (SDS) has the extraordinary physical property of being liquid at room temperature but gelling firmly at 37 degrees C. It has been named 'liquid fibre' and its effect on gastric emptying has been tested in rats and humans, as well as its effect on intestinal distribution in rats. Rats were gavaged with 5 ml of radiolabelled liquid fibre, SDS in water, or water control. Subgroups were killed after 25, 50, 100, 200, and 300 minutes, the gut removed, and the distribution of radioactivity measured scintigraphically. Liquid fibre gelled in the stomach and spread exponentially down the small intestine before 25 minutes. This distribution was maintained for 200 minutes after which the stomach began to empty again. In the human study, 10 healthy men drank 250 ml liquid fibre and placebo labelled with 1.85 MBq technetium tin colloid on separate occasions. Gastric emptying was measured by gamma-camera. Half emptying time significantly increased from 17.7 to 55.8 minutes (means, p < 0.05). The time for 10% to empty (which includes any lag time) increased from 7.0 to 19.4 minutes (p < 0.05). Average emptying rate decreased from 4.49%/min for placebo to 1.60%/min for liquid fibre (p < 0.01). The dramatic delay in gastric emptying suggests liquid fibre may have clinical applications while its liquid formulation should improve acceptability. PMID- 8406150 TI - Is sclerosant injection mandatory after an epinephrine injection for arrest of peptic ulcer haemorrhage? A prospective, randomised, comparative study. AB - A prospective, randomised, comparative study was performed to assess the need for a pure alcohol injection after an epinephrine injection in the arrest of active peptic ulcer bleeding. Sixty four patients with active ulcer bleeding were enrolled in the study. The two groups (epinephrine and epinephrine plus pure alcohol) were matched for sex, age, site of bleed, endoscopic findings, shock, haemoglobin, and concomitant illness at randomisation. The volume of injected epinephrine in the epinephrine and the epinephrine plus pure alcohol groups mean (SD) was 6.0 (3.0) ml and 5.5 (3.0) ml respectively (p > 0.05). The volume of injected pure alcohol in the epinephrine plus pure alcohol group was 1.9 (1.1) ml. Bleeding was initially controlled in 31 (97%) of the epinephrine group and all of the epinephrine plus pure alcohol group. Rebleeding occurred in 11 (36%) of the epinephrine group and in five (16%) of the epinephrine plus pure alcohol group (p > 0.05). Rebleeding was successfully controlled in some patients with treatment by a second injection. Other patients had heat probe thermocoagulation or surgery. Ultimate haemostatic rates were 69% (22/32) and 88% (28/32) for the epinephrine and the epinephrine plus pure alcohol groups respectively (p > 0.05). The epinephrine plus pure alcohol group achieved a better haemostatic effect for spurting haemorrhage (9/10 v 5/11, p < 0.05). The need for emergency operations and blood transfusions were comparable in both groups. The stay in hospital were less in the epinephrine plus pure alcohol group (mean 4.3 v 7.1, p < 0.05). It is concluded that pure alcohol injection after an epinephrine injection can improve the haemostatic rate in patients with spurting haemorrhage and shorten the hospital stay for patients with active bleeding. PMID- 8406151 TI - Inhibition of omeprazole induced hypergastrinaemia by SMS 201-995, a long acting somatostatin analogue in man. AB - Whether the long acting somatostatin analogue SMS 201-995 (octreotide, Sandostatin) could inhibit the basal and meal stimulated hypergastrinaemia and hyperpepsinogenaemia induced by omeprazole was investigated. Eight healthy subjects were randomised to receive five day courses of SMS 201-995 (25 micrograms subcutaneously three times daily), omeprazole (40 mg once a day), a combination of both drugs, or placebo. Basal and meal stimulated serum gastrin and basal serum pepsinogen A and C values were measured the day before treatment, on day five of treatment, and the day after each course of treatment. Omeprazole caused significant increases in basal and meal stimulated peak and integrated serum gastrin values and pepsinogen A and C levels, which were still significantly raised the day after stopping omeprazole treatment. Giving SMS 201 995 with omeprazole significantly reduced any omeprazole induced increases in basal and meal stimulated peak and integrated serum gastrin levels; serum pepsinogen A and C values were significantly inhibited too. Serum gastrin values during combined therapy were not significantly different from those during placebo treatment, whereas pepsinogen A and C levels were still significantly raised. On the day after stopping combined therapy, basal and meal stimulated peak and integrated serum gastrin and serum pepsinogen C (but not pepsinogen A) levels were not significantly different from values obtained on the day after stopping omeprazole alone. SMS 201-995 without omeprazole significantly inhibited basal and meal stimulated peak and integrated serum gastrin levels. Pepsinogen A was also significantly inhibited by SMS 210-995, but the reduction in pepsinogen C failed to reach statistical significance. In conclusion, SMS 201-995 prevents basal and meal stimulated increases in serum gastrin during omeprazole therapy. This finding may have clinical importance in the few patients who have pronounced hypergastrinaemia because of profound long acting acid inhibition. PMID- 8406152 TI - Increased production of luminol enhanced chemiluminescence by the inflamed colonic mucosa in patients with ulcerative colitis. AB - Reactive oxygen species have been implicated as mediators of inflammation in ulcerative colitis. Chemiluminescence is a reliable means of estimating reactive oxygen species in biological media. Increased reactive oxygen species values in the inflamed colonic mucosa in rats were seen by chemiluminescence. The aims of the study were to find out if chemiluminescence is raised in the colonic mucosa of patients with ulcerative colitis and correlates with disease activity, and to elucidate the sources of the chemiluminescence. It was found that reactive oxygen species, as measured by the chemiluminescence technique, are raised in inflamed colonic mucosa and correlates with symptom score, sigmoidoscopic score, disease activity, and activity of the neutrophil enzyme myeloperoxidase. Chemiluminescence was inhibited by a myeloperoxidase inhibitor (azide) and an H2O2 scavenger (catalase) but not by allopurinol, an inhibitor of the enzyme xanthine oxidase. Chemiluminescence was also inhibited by indomethacin, but this did not seem to be related to inhibition of cyclo-oxygenase. These findings suggest that a likely cellular source of reactive oxygen species in the inflamed colon of patients with ulcerative colitis is the neutrophil and that myeloperoxidase conversion of H2O2 to hypochlorous acid, contributes to the chemiluminescence signal and possibly, to the tissue injury. Neither cyclo oxygenase nor lipoxygenase seem to play a part as sources for the chemiluminescence. PMID- 8406153 TI - Initial response and subsequent course of Crohn's disease treated with elemental diet or prednisolone. AB - Elemental diet is as effective as corticosteroids in the treatment of previously untreated Crohn's disease. It is unclear whether a poor nutritional state is a prerequisite for efficacy of elemental diet, whether previously treated patients respond as well, or how duration of remission using elemental diet compares with corticosteroid induced remission. Forty two patients with active Crohn's disease were stratified for nutritional state and randomised to receive Vivonex TEN 2.1 l/day for four weeks, or 0.75 mg prednisolone/kg/day for two weeks and subsequent reducing doses. Nine of 22 (41%) patients assigned to nutritional treatment were intolerant of the diet. Thirty patients completed four weeks treatment. Disease activity decreased on elemental diet from mean (SEM) 4.8 (0.9) to 1.7 (0.6), p < 0.05, and on prednisolone from 5.3 (0.5) to 1.9 (0.6), p < 0.05. For each treatment, nourished and malnourished patients responded similarly. Patients with longstanding disease responded as well as newly diagnosed patients. The probability of maintaining remission at six months was 0.67 after prednisolone, 0.28 after elemental diet, and at one year was 0.35 after prednisolone and 0.09 after elemental diet, p < 0.05. When tolerated, elemental diet is as effective in the short term as prednisolone in newly and previously diagnosed Crohn's disease, and its benefit is independent of nutritional state. The subsequent relapse rate after elemental diet induced remission, however, is greater than after treatment with prednisolone. PMID- 8406154 TI - Neutrophil mucosal involvement is accompanied by enhanced local production of interleukin-8 in ulcerative colitis. AB - The concentration of myeloperoxidase, a neutrophil granule constituent, was measured in the perfusion fluid from sigmoid and rectal segments in patients with ulcerative colitis. The concentrations of myeloperoxidase were increased severalfold in the patients with ulcerative colitis compared with healthy controls pointing to an enhanced neutrophil activity. The release of myeloperoxidase correlated to an enhanced local release of the neutrophil activating peptide interleukin-8 (IL-8). Increased values of tumour necrosis factor (TNF-alpha) were also found during intestinal perfusion of the patients and correlated with those of IL-8. The results obtained are compatible with the hypothesis that local mucosal recruitment/activation of neutrophils in ulcerative colitis is mediated by an enhanced IL-8 synthesis. TNF-alpha may be one relevant factor as a stimulus to IL-8 synthesis. PMID- 8406155 TI - Epidemiology of inflammatory bowel disease in the Province of Granada, Spain: a retrospective study from 1979 to 1988. AB - An epidemiological study of inflammatory bowel disease in the Province of Granada, Spain, was conducted between 1979 and 1988. Altogether, 257 cases were identified: 167 ulcerative colitis, 79 Crohn's disease, and 11 indeterminate colitis. The mean incidence of ulcerative colitis in the 10 year period was 2/10(5) and 0.9/10(5) for Crohn's disease. This is the first epidemiological study in Spain of the incidence of ulcerative colitis and corroborates the results of an earlier population based study on the incidence of Crohn's disease in Spain. PMID- 8406156 TI - In vivo rectal inflammatory mediator changes with radiotherapy to the pelvis. AB - In vivo changes in the rectal values of eicosanoid inflammatory mediators induced by pelvic radiotherapy were measured to study the pathophysiology of the early radiation bowel reaction. Ten patients having pelvic radiotherapy, aged 57 to 78, had rectal dialysis. Values of the eicosanoids leukotriene B4 (LTB4), thromboxane B2 (TXB2), and prostaglandin E2 (PGE2) were measured before radiotherapy, at the end of radiotherapy, and at least four weeks after radiotherapy. Values of LTB4 rose with radiotherapy from 0.21 ng.ml-1 (median) to 1.14 ng.ml-1 (p = 0.012); PGE2 rose from 0.60 ng.ml-1 to 1.58 ng.ml-1 (p = 0.038), and TXB2 rose from 0.365 ng.ml-1 to 1.6 ng.ml-1 (p = 0.005). The rise in eicosanoid inflammatory mediators may have an important role in the pathophysiology of the early radiation bowel reaction. PMID- 8406157 TI - Colonic dysfunction in acute diarrhoea: the role of luminal short chain fatty acids. AB - Faecal concentrations and output of short chain fatty acids (SCFA) were assessed on successive days by gas-liquid chromatography in 24 patients with acute watery diarrhoea. Absorption of water and sodium from the rectum was also measured by a dialysis technique in 17 of these patients and in nine normal subjects in the presence and absence of luminal SCFA. Faecal SCFA concentrations were low on the first day of diarrhoea (mean (SEM) 9.9 (5.8) mmol/kg) and increased to 94.8 (16.4) mmol/kg by the fifth day. Faecal output of SCFA corresponded to these figures. Net water absorption, in the absence of luminal SCFA, was stopped in patients with acute diarrhoea (-59 (81) nl/cm2/min) compared with healthy controls (+322 (63) nl/cm2/min) (p < 0.01). Luminal SCFA restored net water absorption to +184 (67) nl/cm2/min in patients with acute diarrhoea (p < 0.01). Net absorption of sodium decreased in patients with acute diarrhoea in the absence of luminal SCFA, but returned to normal with luminal SCFA. Net secretion of potassium increased in acute diarrhoea, and did not change in the presence of SCFA. Defective absorption from the rectum in acute diarrhoea is reversed by luminal SCFA. The reduction of luminal SCFA in acute diarrhoea treated conventionally may be a factor contributing to colonic dysfunction. PMID- 8406158 TI - Deoxycholate is an important releaser of peptide YY and enteroglucagon from the human colon. AB - Peptide YY (PYY) and enteroglucagon are hormonal peptides found in endocrine cells of the distal intestinal mucosa. Although it is known that plasma concentrations of both peptides increase in response to feeding, the mechanism by which ingested food causes release of colonic hormones is not understood. The release of PYY and enteroglucagon was measured in response to intraluminal stimuli in 176 patients having investigative colonoscopy. Introduction of air, saline (isotonic and hypertonic), glucose (isotonic and hypertonic), oleic acid (without bile salts), and casein hydrolysate all failed to release PYY but glucose caused a small but significant increase in enteroglucagon concentrations. In contrast with the lack of effect of nutrients, infusion of deoxycholic acid produced a rapid and marked dose responsive increase in plasma PYY concentrations when introduced into the sigmoid colon. PYY release was statistically significant at doses between 3.3 mM to 30 mM; for example 10 mM deoxycholate caused a sixfold increase in plasma PYY concentrations. Infusion of 10 mM deoxycholate into the transverse colon or caecum produced an increase of PYY that was similar to the responses in the sigmoid colon. There was also a significant release of enteroglucagon in response to infusion of this bile salt into the sigmoid colon at doses between 3.3 mM and 30 mM. The enteroglucagon response to 10 mM deoxycholate was similar in all three colonic regions. When oleic acid was added to deoxycholate as an emulsion, the release of PYY and enteroglucagon was similar to that seen with the bile salt alone. These findings suggest that bile salts may play an important part in the control of colonic endocrine function and may explain the increased circulating concentrations of colonic regulatory peptides that are seen in malabsorption states and after small bowel resection in humans. PMID- 8406159 TI - Morphometric analysis of intestinal mucosa. V. Quantitative histological and immunocytochemical studies of rectal mucosae in gluten sensitivity. AB - To study changes in rectal mucosa that might be attributable to the effects of gluten, rectal biopsy specimens from untreated and treated gluten sensitised subjects were analysed morphometrically and by immunohistochemical techniques and were compared with a series of disease control mucosae. Although morphometry showed increased populations of plasma cells, lymphocytes, and mast cells in the mucosae of untreated patients, which were reduced (except for mast cells) by dietary gluten restriction, immunohistochemical techniques were far more sensitive in defining these changes. There were highly significant increases in CD3+ and gamma delta+ lymphocytes within both the lamina propria and the epithelium while neutrophils (CD15+ cells) were not at all prominent. Activated (CD25+) lymphocytes expressing interleukin (IL)-2 receptors were increased in lamina propria, usually subjacent to basal lamina, although a few IL-2R+ intraepithelial lymphocytes were found: other IL-2R+ cells were deemed to be macrophages (CD68+). These results clearly indicate that in untreated, gluten sensitised subjects the rectal mucosa shows a lymphoplasmacytoid reaction that is responsive to gluten restriction. The absence of neutrophilia suggests that this lesion is not a conventional inflammatory type proctitis, but rather one presumed to be induced by gluten antigen(s) present in the faecal stream--that is, a cell mediated form of response. PMID- 8406160 TI - Selective affective biasing in recognition memory in the irritable bowel syndrome. AB - The cognitive model of depression assigns a central role to negatively biased information processing in the pathogenesis of the emotional disorder. The relationship between depression and irritable bowel syndrome (IBS) was explored from a cognitive perspective. A word recognition memory task was constructed: subjects had to memorize and subsequently recognise a set of emotionally loaded stimulus words with either positive, neutral, or negative connotations. Four age matched groups participated--30 IBS patients, 28 depressed patients, 28 patients with organic gastrointestinal disease, and 30 healthy volunteers. The depressed patients, as would be expected, showed a significant bias in favour of emotionally negative words (p < 0.05): the IBS patients showed the same negative bias. In addition the IBS patients made significantly more false-positive type errors in recognising emotionally negative words than either the depressed patients (p < 0.05) or the healthy volunteers (p < 0.01). This suggests that the IBS patients have a peculiar confirmatory bias for negative material. This may have clinical relevance in terms of the IBS patients' evaluation of their own abdominal sensory experience. PMID- 8406161 TI - Xenobiotic metabolising enzyme expression in colonic neoplasia. AB - The cytochrome P450, epoxide hydrolase, and glutathione S-transferase enzyme families play an important part in the metabolism of many carcinogens and anti cancer drugs. The expression of two forms of cytochrome P450 (P450 1A and P450 3A), epoxide hydrolase and of the alpha, mu, and pi forms of glutathione S transferase in normal colon, colonic adenomas, and adenocarcinoma of the colon were studied by immunohistochemistry. This allowed the precise cellular site and distribution of each enzyme to be determined. Expression of all the xenobiotic metabolising enzymes studied was almost wholly confined to the epithelial cells, whether in normal, adenoma or carcinoma samples, except that cytochrome P450 3A was also identified in mast cells and glutathione S-transferase pi was also present in chronic inflammatory cells. Cytochrome P450 was present in only a small proportion of normal colon samples, whereas epoxide hydrolase and glutathione S-transferase mu were identified in about half, and glutathione S transferase alpha and pi in most normal samples. By contrast all the enzyme forms studied were expressed in virtually all adenomas and in over half the carcinomas. These results suggest that cytochrome P450 1A and cytochrome P450 3A are more specific markers of colonic neoplasia than epoxide hydrolase or glutathione S transferases alpha, mu, and pi. PMID- 8406162 TI - Nuclear DNA content of isolated crypts of background colonic mucosa from patients with familial adenomatous polyposis and sporadic colorectal cancer. AB - The DNA content of the upper one third of the crypt epithelium was compared with that of the lower two thirds in the background colorectal mucosa of eight cases of familial adenomatous polyposis (FAP) and eight control cases of sporadic colorectal cancer (SCRC). Intact crypts were isolated by incubating fresh lesion free colorectal mucosa in calcium and magnesium free Hanks' balanced salt solution (CMFH) containing 30 mM EDTA for 30 minutes at 37 degrees C and then agitating in CMFH. The crypts were then separated from the lamina propria, fixed in 70% ethanol and under a dissecting microscope divided manually into upper and lower portions. Each portion was digested with pepsin to obtain a suspension of single nuclei, and smears of the nuclei were stained with 4',6,-diamidino-2 phenylindole dihydrochloride (DAPI). Nuclear DNA was determined using a cytophotometric microscope. Results showed that the DNA content of the epithelium of the upper one third of crypts was diploid in both FAP and SCRC cases, and that proliferative fractions with diploid peaks were present in the lower two thirds of the crypts in both groups. These results support our previous finding that the proliferative compartment of background crypts is confined to the lower two thirds and does not extend to the upper parts of the crypts. PMID- 8406163 TI - Bile duct injury after laparoscopic cholecystectomy: the value of endoscopic retrograde cholangiopancreatography. AB - This study describes the value of endoscopic retrograde cholangiopancreatography (ERCP) in patients with bile duct injury after laparoscopic cholecystectomy. Twelve consecutive patients were studied over a one year period. In all patients the biliary tree was visualised during ERCP. Four patients had complete bile duct obstruction, seven patients had a stricture (two with concomitant leakage), and one patient had leakage from a hepatic branch. Three patients with complete obstruction, presented with a relatively prolonged symptom free, 'silent' period before diagnosis. In all four patients with complete transection, a proximal hepaticojejunostomy was performed. In one patient with a tough fibrous stricture, secondary to incorrect clip placement, passage of the guidewire was impossible, leaving surgical reconstruction as the only therapeutic option. All remaining seven patients with leakage or strictures, or both were successfully treated by endoscopic sphincterotomy only (n = 1) or sphincterotomy and subsequent stent placement (n = 6). When patients do not recover uneventfully after laparoscopic cholecystectomy even without cholestasis or jaundice, early ERCP is recommended as a safe and valuable method to detect bile duct injury and to suggest treatment. Subsequently, more than half of such patients can be treated endoscopically. Extended follow up is needed to evaluate the longterm results. PMID- 8406164 TI - Pancreatitis in Finland between 1970 and 1989. AB - The incidence and mortality from pancreatitis in Finland between 1970 and 1989 were studied and compared with the alcohol consumption in the country and with the incidence of liver cirrhosis and gall stone disease. Hospital discharge data were obtained from the Finnish National Agency for Welfare and Health, the causes of deaths from the Finnish State Statistics, and annual alcohol consumption from the Finnish State Alcohol Company. There were 56,353 hospital treatment periods because of pancreatitis. The incidence of pancreatitis discharges increased from 46.6 to 73.4/100,000/year. In men it increased from 59.1 to 113.4, but in women it remained unchanged (mean 35.0). The incidence of pancreatitis discharges correlated with the alcohol consumption in Finland (r = 0.78, p = 0.0001). The incidence of pancreatitis discharges correlated in men, but not in women, with the incidence of liver cirrhosis (r = 0.81, p = 0.0001). In women, but not in men, the incidence of pancreatitis discharges correlated with the incidence of gall stone disease discharges (r = 0.77, p = 0.0001). The incidence of discharges due to haemorrhagic pancreatitis and pancreatic abscess doubled in men and remained unchanged in women. Pancreatitis death rate decreased from 5.9% (men 4.8%, women 7.0%) to 2.6% (men 2.4%, women 2.7%). PMID- 8406165 TI - Reversible bilateral ureteric obstruction due to a pancreatic pseudocyst. AB - An unusual case of bilateral ureteric obstruction and hydronephrosis due to pancreatic pseudocyst formation, after an episode of acute pancreatitis is reported. All abnormalities resolved with conservative management. Possible reasons for such ureteric obstruction include periureteric fat necrosis by pancreatic enzymes and compression by the inflammatory mass. PMID- 8406166 TI - Iatrogenic pancreatitis in familial adenomatous polyposis. AB - The first case of a patient with familial adenomatous polyposis (FAP) who developed pancreatitis after routine screening and biopsy of the ampulla of Vater is described. PMID- 8406167 TI - Thrombosis of splenic artery pseudoaneurysm complicating pancreatitis. AB - The natural history of pseudoaneurysms complicating pancreatitis is unknown. A patient with chronic pancreatitis is described in whom thrombosis of a splenic artery pseudoaneurysm occurred. Early diagnosis and radical treatment of a bleeding pseudoaneurysm are mandatory. When elective treatment is considered, however, contrast enhanced computed tomography may be useful just before surgery as thrombosis may occur. PMID- 8406168 TI - Carcinoid tumour of the gall bladder: two case reports and a review of published works. AB - This paper reports on two patients with carcinoid tumours of the gall bladder who presented to the Aberdeen hospitals during the period 1970 to 1990 and reviews all previously reported cases in published works. PMID- 8406169 TI - Gall stone recurrence and its prevention: the British/Belgian Gall Stone Study Group's post-dissolution trial. AB - The British/Belgian Gall Stone Study Group (BBGSG) post-dissolution trial was a prospective, multicentre, randomised, double blind trial of: (i) low dose ursodeoxycholic acid, (ii) placebo, and (iii) a high fibre, low refined carbohydrate diet in the prevention of gall stone recurrence in patients with complete gall stone dissolution. Further aims included establishing the timing and frequency of recurrence and its association with biliary symptoms, a comparison of the sensitivity of ultrasonography v oral cholecystectography in detecting recurrent stones, and a search for risk factors predicting recurrence. Ninety three patients entered the study, and 82 were followed up for up to five years (mean (SEM) 28 (1.5) months) with six monthly ultrasonography and yearly oral cholecystectography. There were 21 recurrences (26 by oral cholecystectography or ultrasonography, or both), only two of which were symptomatic, which were detected between 12 and 42 months after trial entry. This corresponded to an actuarial recurrence rate of 33.9 (7.0%) by lifetable analysis at 42 months and subsequently. There were four recurrences in the ursodeoxycholic acid, six in the placebo, and 11 in the diet groups, corresponding to 21.9 (9.9)%, 27.4 (10.1)%, and 45.8 (12.4)% respectively at 42 months by lifetable analysis (NS). Variables including age, obesity, menopausal state, pregnancy, and oestrogen containing drugs were not shown to affect recurrence rate. Men had more frequent recurrence than women (NS). Patients who had had multiple stones experienced more recurrences than did those with single stones (NS). Recurrence did not occur in patients who took non-steroidal anti-inflammatory drugs (NSAIDs) (p < 0.02). The stone free interval between stone dissolution and trial entry proved to be important--those stone free > nine months had a recurrence rate of only 12.7 (6.0)% at 42 months compared with 55.4 (12.5)% in those stone free < nine months (p < 0.01). There was imbalance between the ursodeoxycholic acid and placebo groups for this factor, and after applying a statistical correction, the adjusted recurrence rate in the ursodeoxycholic acid group was 15% compared with 30% in both placebo and diet groups (NS). These data suggest that after medical dissolution, the risk of gall stone recurrence is not reduced by a high fibre, low refined carbohydrate diet: it may be lowered, but not abolished, by low dose ursodeoxycholic acid. PMID- 8406170 TI - Radioisotope determination of regional colonic transit in severe constipation. PMID- 8406171 TI - Polyunsaturated fatty acid pattern and fish oil treatment in inflammatory bowel disease. PMID- 8406172 TI - Antibodies to Mycobacterium paratuberculosis. PMID- 8406173 TI - Extracorporeal shock wave lithotripsy and gall bladder stones. PMID- 8406174 TI - Screening and management of familial adenomatous polyposis. PMID- 8406175 TI - Searching for the old. PMID- 8406176 TI - Interventions for sleep pattern disturbances. PMID- 8406177 TI - Caring for the confused geriatric surgical patient. AB - Postoperative confusion in elderly patients is generally a temporary and reversible condition. The causes and contributing factors are numerous. The number of elderly surgical patients will continue to increase as this age group continues to grow. Our role as nurses is to provide the care that will not only focus on early recognition, assessment, and treatment of postoperative confusion, but will also recognize factors that preoperatively place a patient at increased risk and provide care that will prevent or minimize postoperative confusion. PMID- 8406178 TI - Effects of age on cardiovascular functioning. With an understanding of the changes that occur in the cardiovascular system, and of the risk factors to adequate functioning, nurses will be more able to plan interventions. PMID- 8406179 TI - Distal bypasses in patients over age 75. With a thorough medical evaluation, patients over 75 in need of distal bypass may not be at greater risk than younger patients. PMID- 8406180 TI - Nursing assessment of mental status in the elderly. Formal mental status testing is a tool gerontological nurses should put to use. PMID- 8406181 TI - Factors influencing retention of directors of nursing at rural long-term care facilities. What difficulties do rural LTC facilities face in attracting and retaining qualified NA/DONs? PMID- 8406182 TI - The efficient recording of diabetic care. The nurse in an extended care facility needs an "at-a-glance" recording system to assess diabetic clients' blood glucose levels and other telltale symptoms of hypoglycemia. PMID- 8406183 TI - Understanding Huntington's disease: an overview of symptomatology and nursing care. Nurses must balance safety, quality-of-life issues, the need for personal choice, and the meaning of human dignity when caring for patients with Huntington's disease. AB - Patients and families who have been affected by HD deserve high-quality and dignified nursing care. The complexity and extent of losses and uncertainties imposed by this illness challenge nursing. We have briefly reviewed the current state of knowledge regarding the illness and the nursing interventions appropriate for working with people with HD. What is missing is an empirically validated body of interventions to guide nursing practice. Nursing research is needed to validate existing care practices, generate new interventions, and to begin to build theoretical explanations that assist nurses in providing care to people with Huntington's disease. PMID- 8406184 TI - A celebration of cultures in a long-term care facility. A day of cultural awareness may help staff and residents forge stronger relationships through greater understanding. AB - Cultural interaction is older than the spice route. Groups of people require little encouragement to share "how we do things back home," as well as favorite foods, customs, and beliefs. This interchange is important in decreasing myths and stereotypes that occur between cultures. Identification of differences is an essential as the discovery of how people are alike. The acknowledgement of sameness promotes unity, but recognition of differences promotes cooperative effort. There is enormous strength in cultures working together, supplementing the limitations and talents of each other. This celebration of cultures was our attempt to encourage staff-resident interactions and strengthen relationships among staff, residents, and all departments. Although several years have passed, staff still speak of Cultural Awareness Day as having been a positive force in promoting good relationships. The day provided entertainment and valuable lessons for all. PMID- 8406185 TI - Making your nutrition education efforts worthwhile. PMID- 8406186 TI - Sex steroid receptors in endometrial carcinoma. PMID- 8406187 TI - Steroid receptor concentrations in endometrial carcinoma: effect on survival in surgically staged patients. AB - Estrogen and progesterone receptor concentrations were measured in the primary tumors of 137 surgically staged women with clinical stages I and II endometrial carcinoma. For each steroid, increasing receptor concentrations were associated with a decrease in hazard (increase in survival) and the effect was linear for each receptor. When expressed dichotomously, steroid receptor status was also significantly associated with a number of known risk factors, and the significance of the association was influenced by the receptor concentration used as the criterion for receptor positivity. In a multivariate analysis, only progesterone receptor concentration affected survival independently, but the effect disappeared when the analysis was restricted to women with disease confined to the uterus. We conclude that the estrogen and progesterone receptor status of the primary tumor is of limited prognostic significance in endometrial carcinoma unless extrauterine disease is present. PMID- 8406188 TI - Consolidation intraperitoneal chemotherapy in epithelial ovarian cancer patients following negative second-look laparotomy. AB - Fifty-six patients with Stage III ovarian carcinoma who had a negative second look laparotomy following optimum cytoreduction, cisplatin, and cyclophosphamide chemotherapy were enrolled in a pilot study to receive consolidation intraperitoneal (IP) chemotherapy. Forty-one patients received three courses of IP cisplatin (80 mg/m2). Fifteen patients with cisplatin-induced toxicity received three courses of IP mitoxantrone (10 mg/m2). Toxicity was minimal, with only 2 patients suffering peritonitis and 1 patient developing an enterotomy at the time of catheter removal. Median follow-up for the IP cisplatin and IP mitoxantrone group has been 24 and 30 months, respectively. Ten of the 41 patients (24%) who received IP cisplatin and 4/15 (26%) of the IP mitoxantrone patients have recurred (P = ns). Median time to recurrence was 18 months for both groups. This study indicates that consolidation intraperitoneal chemotherapy following negative second-look laparotomy is feasible with either cisplatin or mitoxantrone. However, many of these patients still recurred and prospective trials will be required to determine which (if any) consolidation intraperitoneal treatment is best following a negative second-look laparotomy. PMID- 8406189 TI - The Groshong catheter as an intraperitoneal access device in the treatment of ovarian cancer patients. AB - With the development of new intraperitoneal treatments in ovarian cancer, safe and convenient access to the peritoneal cavity is now required. This report reviews the University of Alabama at Birmingham's experience with the Groshong catheter as an intraperitoneal access device. The Groshong was easily inserted intraperitoneally in 20 ovarian cancer patients and used to deliver 81 courses of intraperitoneal therapy over 2310 patient-days. There were no catheter-related complications during treatment and only one exit site infection after catheter removal. Further investigation of the Groshong catheter as a novel intraperitoneal access device appears warranted. PMID- 8406190 TI - Importance of serum sialic acid and lactate dehydrogenase in diagnosis and treatment monitoring of cervical cancer patients. AB - In an attempt to establish a blood-based biochemical index for diagnosis of cervical cancer and treatment monitoring of patients suffering from the disease, serum levels of total sialic acid (TSA), lipid-bound sialic acid (LSA), and lactate dehydrogenase were estimated by highly specific spectrophotometric methods. Serum concentrations of the markers in 108 untreated cervical cancer patients were compared with the levels of the biomarkers in 125 healthy, age matched female individuals (controls). The alterations in serum levels of the markers after radiotherapy in cervical cancer patients were also observed. The levels of all markers were significantly higher (P < 0.001) in untreated cervical cancer patients compared to the controls. TSA was found to be the most sensitive (90.74%) marker for diagnosis of cervical cancer. Combined use of the markers revealed maximum (100%) sensitivity. In comparison between early (stage I+II) and advanced (stage III+IV) malignant disease, the markers showed insignificant changes. TSA and LSA values in patients who did not respond to radiotherapy were significantly higher (P < 0.05 and P < 0.001, respectively) than those of the responders. The results suggest that combined evaluation of the markers is helpful for diagnosis as well as for treatment monitoring of cervical carcinoma patients. PMID- 8406191 TI - Plasma methotrexate levels in patients receiving etoposide, methotrexate, actinomycin, cyclophosphamide, and vincristine (EMA-CO) chemotherapy for trophoblastic neoplasia. AB - Methotrexate serum levels were measured in eight patients receiving etoposide, methotrexate, actinomycin, cyclophosphamide, and oncovin (EMA-CO) chemotherapy for high-risk and actinomycin-resistant gestational trophoblastic neoplasia. During 41 courses of EMA there were only 3 instances where the level of methotrexate was greater than 9 x 10(-7) M/liter at 24 hr postinfusion. A level higher than this may be associated with subsequent toxicity. Use of postmethotrexate leucovorin rescue should continue, but routine measurement of methotrexate with EMA-CO is unnecessary unless there is compromise of renal or hepatic function. PMID- 8406192 TI - Flow cytometry as a prognostic indicator in women with borderline epithelial ovarian tumors. AB - Flow cytometry has been used to rapidly and reliably measure DNA content in malignant tumor cells. Although several studies have suggested that DNA ploidy is a powerful predictor of survival in women with epithelial ovarian cancer, few have determined the usefulness of this procedure in women with borderline tumors. Using data from a population-based tumor registry covering all of western Washington State, women who died prior to 1992 as a consequence of developing a borderline ovarian tumor between 1975 and 1986 were compared to an age, histology, and histologic stage-matched sample of women with the same diagnosis still living after at least 5 years of follow-up. Flow cytometry analysis was conducted using sections of tumor from the original paraffin blocks. Overall, 25% of the women who died and 24% of those still alive had aneuploid DNA tumors (odds ratio = 1.2, 95% CI = 0.3-4.9). This lack of association stands in contrast to the strong relationship of aneuploid status to mortality in an earlier, similarly designed, study of borderline ovarian tumors. We believe that additional studies are required prior to concluding that the clinical course of women with borderline tumors can be predicted by the ploidy status of their tumor's DNA. PMID- 8406193 TI - Adenosquamous carcinoma of the cervix: prognosis in early stage disease treated by radical hysterectomy. AB - The prognosis of adenosquamous carcinoma of the cervix compared to the pure cell types of this disease is a controversial issue. Survival rates vary widely among published series, with some authors finding the prognosis to be much worse and others finding it to be equal. We have studied a group of 290 patients, all of whom had the diagnosis of stage IB or IIA cervical cancer, and all of whom were treated by radical hysterectomy and bilateral pelvic and paraaortic lymphadenectomy. The pathology specimens were reviewed for every case. Median follow-up for all living patients was 73 months. Forty-five patients (15.5%) had adenosquamous histology, 220 (75.9%) had squamous cell, and 25 (8.6%) had adenocarcinoma. By X2, there was no significant difference among the three groups with regard to race, economic status, number of grade 3 lesions, number with positive pelvic lymph nodes, number with positive margins, stage distribution (IB vs IIA), or number of recurrences. Using Student's t test, there was no significant difference between any combination of two groups with regard to mean patient age, mean depth of invasion, or mean tumor size. Estimated disease-free survival and overall survival were not different among the three cell types. We conclude that for early stage cervical cancer treated by radical hysterectomy, the adenosquamous cell type does not carry a worse prognosis than either of the pure cell types. PMID- 8406194 TI - Epithelial ovarian tumors of borderline malignancy: long-term follow-up. AB - Thirty-nine patients underwent primary surgery for epithelial ovarian tumors of low malignant potential at the Massachusetts General Hospital between 1970 and 1980. Eighty-five percent of patients were found to have Stage I disease and 15% were found to have Stage III disease. Fifty-four percent of patients had a tumor with serous histology, 39% had a tumor with mucinous histology, and the remainder of patients had tumors with an endometrioid or mixed-cell type. Second malignancies and benign ovarian tumors were frequently found concomitantly with the borderline tumors or in follow-up. Gastrointestinal and endometrial adenocarcinomas were the most common second malignancies and were frequently found associated with a borderline tumor of serous histology. Follow-up was available in all 39 patients (100%). Mean time of follow-up was 11.8 years. Sixty nine percent of patients are clinically without evidence of disease with a mean follow-up of 14.7 years, 23% died of other causes, 5% died of disease, and 3% died with disease and sepsis. All patients dying with disease did so within 7.3 years of their primary surgery. Seven patients underwent conservative surgery, defined as preservation of some ovarian tissue. Six of 7 patients are clinically free of disease with a mean follow-up of 14.6 years; 1 patient died of other causes. No patients treated conservatively had a recurrence of their disease. PMID- 8406195 TI - The significance of histology and morphometry in predicting lymph node metastases in patients with squamous cell carcinoma of the vulva. AB - The material consists of a series of 73 patients with squamous cell carcinoma of the vulva. The site and the size of the primary tumor and the histological status of the lymph nodes of the groin were known. Two pathologists evaluated nuclear hyperchromatism, nuclear polymorphism, histological differentiation, number of mitoses, inflammatory response, and vascular invasion and graded these parameters from one to three. The reliability of the histopathological grades evaluated by the kappa coefficient showed considerable interobserver variation. Despite this a model which included the subjective parameter nuclear hyperchromatism could predict patients without lymph node metastases. The model consisted of patients with tumors which were not situated on the clitoris, were less than 40 mm in diameter, and exhibited only slight hyperchromatism. The model fitted 19 (26%) and 14 (19%) of the patients with two different pathologists evaluating the nuclear hyperchromatism and none of these patients had lymph node metastases. The quantitative parameter--mean nuclear volume--determined by morphometry was of no diagnostic value for the prediction of patients without groin node metastases at the time of operation. PMID- 8406196 TI - Pelvic and parametrial lymph nodes in the quality control of the surgical treatment of cervical cancer. AB - Between 1971 and 1989 a total of 420 patients underwent radical abdominal hysterectomy with pelvic lymphadenectomy at our hospital for stage Ib, IIa, or IIb cervical cancer. The entire lymph node material was processed in serial sections and stained with hematoxylin and eosin. Lymph nodes were counted and the sizes of metastases measured. The entire surgical specimen was fixed as a whole with the parametria spread out. The size of the tumor was measured by morphometry. Cases treated between 1971 and 1979 were compared with those treated between 1980 and 1989. The median number of pelvic lymph nodes removed per patient was 24 between 1971 and 1979 and 35 between 1980 and 1989 (P = 0.0001). Significantly more nodes were removed at each node group (P = 0.01). Between 1971 and 1979 no common iliac nodes were obtained in 56 patients and no left common iliac nodes in 74 patients, compared to only 1 and 6 patients, respectively, between 1980 and 1989. The rate of patients with positive lymph nodes was 33% (63/195) between 1971 and 1979 and 55% (101/225) between 1981 and 1989 (P = 0.008). In the first study period the median number of parametrial lymph nodes was 2 compared to 3 in the second period. The rate of patients with positive parametrial lymph nodes increased from 15 to 24% (P = 0.027). The results of this review indicate that exacting morphologic processing of the entire lymphatic tissue obtained at surgery permits accurate postoperative staging and assessment of risk factors for decisions on adjuvant treatment. Histologic evaluation objectifies the radicality of the procedure and is useful in the quality control of the surgical treatment of cervical cancer. PMID- 8406197 TI - Histologic correlates of progression-free interval and survival in ovarian clear cell adenocarcinoma. AB - Unlike other ovarian epithelial cancers, histologic measures of differentiation of ovarian clear cell adenocarcinoma have not been found to be reliable predictors of progression-free interval and survival. Forty-two consecutively treated patients with pure ovarian clear cell adenocarcinoma were identified and all histologic materials were reviewed. The following histologic features were assessed: architectural pattern (tubular/tubulocystic, papillary, solid, and mixed), average and maximal mitotic activity per 10 high-power fields (hpf's), percentage of cells with clear cytoplasm, and nuclear grading (both Fuhrman et al.'s (Am. J. Surg. Pathol. 6, 655-663, 1982) and Christopherson et al.'s (Cancer 49, 1511-1523, 1982) methods). Surgical stage was most predictive of survival (P < 0.01) and subsequent comparisons using Cox proportional hazards modeling were adjusted for Stages III and IV. Greater than six mitotic figures per 10 hpf's predicted a poorer survival (P = 0.05) but was not predictive of progression-free interval (P = 0.28). Survival and progression-free interval in patients with tumors of mixed architectural pattern tended to be shorter than those for patients where one type did predominate, but this was not statistically significant (P = 0.06). The tubulocystic pattern was not predictive of patient outcome nor was either method of nuclear grading. Stage and average mitotic activity seem to be the best predictors of survival for patients with ovarian clear cell adnocarcinoma, while architectural pattern and nuclear grading are not as reliable. PMID- 8406198 TI - Expression of ras p21 oncoprotein in normal and neoplastic human endometrium. AB - The ras-encoded p21 protein expression was investigated in 18 normal endometrial tissues and in 37 human primary endometrial carcinomas by Western blotting analysis. Scattered p21 levels were found in normal specimens (mean = 1.29 OD; median = 1.10 OD; range = 0.33-2.65). The p21 levels were significantly higher in secretory (mean = 1.99 OD; median = 2.16 OD; range = 0.71-2.65) than in proliferative (mean = 0.97 OD; median = 1.07 OD; range 0.38-1.73) endometrium (P = 0.009) and higher in primary endometrial carcinomas (mean = 2.05 OD; median = 2.04 OD; range 0.21-4.36) than in normal proliferative tissues (P = 0.004). Immunohistochemical analysis showed that most of the tumor cells expressed p21 oncoprotein while the stromal component was unreactive. No correlation between p21 expression and histopathological characteristics of the tumors was observed. Moreover, estrogen receptor (ER)-positive tumors expressed higher p21 levels than did ER-negative tumors (77% vs 33%; P = 0.009). A similar trend, although not statistically significant, was found between p21 values and progesterone receptor expression (74% vs 44%; P = 0.060). On the other side, p21 levels were unrelated to epidermal growth factor receptor levels. Further studies should verify the possible significance of p21 expression in the prognostic characterization of patients with endometrial cancer. PMID- 8406199 TI - Extraovarian peritoneal serous papillary carcinoma: a case-control retrospective comparison to papillary adenocarcinoma of the ovary. AB - Since the establishment of extraovarian peritoneal serous papillary carcinoma (EPSPC) as a clinical entity in 1959, less than 250 cases have been described and its clinicopathologic features remain obscure. The present series is a retrospective, case-controlled study comparing the response and survival to cytoreductive surgery followed by cisplatin-based multiagent chemotherapy of 33 women with confirmed EPSPC versus 33 cases with papillary serous ovarian cancer (PSOC). Each EPSPC case was matched to a PSOC control for extent and distribution of disease prior to and following cytoreductive surgery, tumor grade, patient age, and treatment. Additionally, the new Gynecologic Oncology Group criteria for the diagnosis for EPSPC are discussed. There were no significant differences in tumor response to therapy, disease-free interval, and actuarial survival between cases and controls. These data suggest that EPSPC is clinically similar to PSOC and support the need for a prospective clinical trial to compare these two entities further. PMID- 8406200 TI - Advanced ovarian carcinoma diagnosed during pregnancy in a patient with human immunodeficiency virus infection. AB - Many malignancies appear to occur with increased frequency and aggressive patterns of spread in patients seropositive for human immunodeficiency virus (HIV). The relationship between HIV infection and cervical neoplasia suggests that these patients present with more advanced disease and demonstrate poor response to therapy. To date, there have been no reported cases of ovarian cancer with concomitant HIV infection. We describe a young, gravid woman with an advanced ovarian carcinoma diagnosed at the time of delivery. Following poor response to cytoreductive surgery and initial chemotherapy, she was found to be HIV-seropositive. She received multiple chemotherapeutic regimens and experienced significant complications associated with her treatment and HIV infection. She progressively deteriorated and died within 13 months of diagnosis. Based on these findings and experience with other HIV-associated malignancies, it is apparent that the conventional approach to therapy is inadequate to treat the advanced and more aggressive form of disease seen in women infected with HIV. PMID- 8406201 TI - Uterine angiomyolipoma associated with pregnancy. AB - Uterine angiomyolipomas are rare lesions composed of mature adipose tissue, smooth muscle, fibrous connective tissue, and blood vessels in varying proportions. We reported the first case of angiomyolipoma associated with a normal pregnancy. Initially, the tumor developed intramurally and could have been confused with a partial molar gestation. After delivery, tumor development was extensive and subserosal, making differential diagnosis from a sarcoma difficult. The question of histological diagnosis, as well as that of immunocytochemical analysis which seems to be helpful in such cases, is discussed here. PMID- 8406202 TI - Serous papillary carcinoma of the cervix. AB - Two cases of serous papillary carcinoma of the cervix are reported. Both patients had clinical stage IB disease. One patient underwent radical hysterectomy and bilateral pelvic lymphadenectomy. The second patient was treated with pelvic radiation therapy after exploration demonstrated uterosacral ligament metastasis. Both patients are alive at 35 months after treatment with local therapy alone. In contrast to ovarian, tubal, and endometrial serous carcinomas, local therapy appears to be effective treatment for serous carcinomas of the cervix. PMID- 8406203 TI - Ureteroarterial fistulae in exenteration patients with indwelling ureteral stents. AB - Ureteroarterial fistulae (UAF) are a rare complication of prolonged ureteral stenting. To date only three patients have been reported who have developed UAF after pelvic exenteration. This report presents two additional patients with UAF following exenteration and prolonged ureteral stenting. Rapid diagnosis with pelvic arteriography and retrograde ureteral angioplasty balloon catheter placement is discussed, and successful management with femoral artery embolization and bypass surgery is reviewed. PMID- 8406204 TI - Peritoneal papillary serous carcinoma in a woman with a history of utero DES exposure. AB - Antenatal exposure to diethylstilbestrol (DES) has been implicated in the development of clear-cell adenocarcinoma of the vagina and cervix. In addition, there are a handful of case reports of gram cell tumors and other benign ovarian tumors in the offspring of women treated with DES during pregnancy. This the first report of a papillary serous carcinoma of the peritoneum following in utero exposure to DES. As the population of women with a history of in utero exposure to DES is reaching into the fourth decade, these patients need to be closely monitored for the development of gynecologic malignancies, especially epithelial and germ cell ovarian tumors. We also encourage the reporting of these tumors of the DES registry to document the exact incidence of these malignancies. PMID- 8406205 TI - Management of advanced ovarian epithelial cancer in the renal transplant patient. AB - Stage IIIC, grade 1 papillary serous adenocarcinoma of the ovary was diagnosed in a 28-year-old renal transplant recipient. She had been treated with the immunosuppressive agents azathioprine and methylprednisolone for 7 years prior to the discovery of the ovarian cancer. Surgical excision of the tumor was suboptimal due to involvement of the allograft; however, the patient achieved a complete clinical response after eight courses of cisplatin and cyclophosphamide. Since multiagent immunosuppressant therapy may have contributed to the development of the ovarian carcinoma, the intensity of immunosuppression was decreased by discontinuing the azathioprine as soon as the diagnosis of ovarian cancer was made. The methylprednisolone, however, was continued to decrease the possibility of organ rejection. After completion of chemotherapy, the patient was started on a daily regimen of low-dose oral cyclophosphamide as an immunosuppressant. Four months following the completion of cytotoxic therapy, she developed clinically evident disease in the pelvis. Subsequent salvage therapy with carboplatin failed. The patient died from progressive disease 26 months after initial diagnosis. She never developed evidence of renal rejection. Combined modality cancer therapy, preservation of allograft function, and modification of immunosuppressant therapy are important goals in the renal transplant patient with advanced ovarian carcinoma. PMID- 8406206 TI - Primary breast cancer of the vulva. AB - Supernumerary breasts on the vulva are a rare occurrence. There have been 26 cases previously reported and only 4 case reports of primary mammary carcinoma of the vulva. This is the fifth case report of mammary carcinoma of the vulva and the second report using tamoxifen for adjuvant treatment. PMID- 8406207 TI - Spontaneous perforation of pyometra presenting as generalized peritonitis in a patient with cervical cancer. AB - Pyometra is a relatively common event. However, it is extremely rare for this condition to perforate spontaneously into the peritoneum, and to date only 14 cases have been reported in English literature. We recently experienced such a case in a patient with cervical cancer. The clinical features, pathologic findings, diagnosis, therapy, and outcome of these 15 cases were reviewed. Only 5 cases were associated with malignant diseases, consisting of 2 cervical cancers, 2 colon cancers, and 1 endometrial cancer. All of the patients developed generalized peritonitis, and 4 patients died of this. Correct preoperative diagnosis was difficult. Therefore, immediate and appropriate therapy is important. PMID- 8406208 TI - Non-Hodgkin's lymphoma presenting as a urethral caruncle. AB - Primary malignant lymphoma of the female urethra is a rare entity. Five cases of non-Hodgkin's lymphoma of the female urethra have been previously reported. We report another case presenting with a urethral caruncle. A review of the literature is included. PMID- 8406209 TI - [Vulvar consultation]. PMID- 8406210 TI - [Current nomenclature of vulvar diseases]. PMID- 8406211 TI - [Vulvar changes in childhood and adolescence]. PMID- 8406212 TI - [Symptom specific differential diagnosis of vulvar diseases]. PMID- 8406213 TI - [Current diagnosis and therapy of inflammatory vulvar diseases]. PMID- 8406214 TI - [The significance of viral disease of the vulva]. PMID- 8406215 TI - [Premalignant vulvar lesions]. PMID- 8406216 TI - [Skin diseases of the vulva]. PMID- 8406217 TI - [Psychosomatic aspects of vulvar manifestations]. PMID- 8406218 TI - [Principles of surgical treatment of vulvar cancer and precancerous lesions]. PMID- 8406219 TI - [Bloody urine noted in the course of hysterectomy]. PMID- 8406220 TI - Media messages, empathy, and social work. PMID- 8406221 TI - Reaction of Vietnam veterans to the Persian Gulf War. AB - The notion that veterans' war experiences can be reawakened on exposure to subsequent wars has not received the attention it merits by mental health professionals. A current or recent war can significantly affect veterans; in particular, some Vietnam veterans have had intense reactions to the Persian Gulf War. This article reviews the evolution of the concept of combat-related posttraumatic stress disorder and analyzes reports of Vietnam veterans' reactions to the Persian Gulf War. A case study of a Vietnam veteran whose traumatic memories were reawakened with the onset of the Persian Gulf War is presented, and implications for social work practice are discussed. PMID- 8406222 TI - Stress management consultation to Israeli social workers during the Gulf War. AB - Social workers operating under emergency conditions may feel overwhelmed and incapacitated by their own stress reactions. During the Gulf War, social work faculty in Israel acted as consultants to help hospital social workers manage stress and regain professional efficacy. The Stress Management Consultation (SMC) was a short-term group intervention designed to enable social workers to work through their stress reactions and to model a method the workers could use with their own target populations. Qualitative feedback from both participants and administrators indicated that the SMC model was effective and could be applicable to a large range of settings and treatment populations. PMID- 8406223 TI - Psychological traumas and depression in a sample of Vietnamese people in the United States. AB - This article investigates the relationships among premigration stresses, nightmares, acculturation stresses, personal efficacy, and depression in a sample of 147 adult Vietnamese Americans. The analysis revealed that premigration stresses, nightmares, and acculturation stresses had significant indirect effects on depression. Acculturation stresses diminish personal efficacy, and a weakness of personal efficacy leads to higher depression. Age, gender, marital status, and English language ability also exert differential effects on premigration stresses, nightmares, acculturation stresses, personal efficacy, and depression. Health care professionals, social workers, health care organizations, and social work researchers who work with Vietnamese clients should be more culturally sensitive when planning and implementing services and developing research instruments. PMID- 8406224 TI - Support groups as open systems: a model for practice and research. AB - Despite the increasing of use of support groups to respond to the needs of people dealing with crises, life transitions, and chronic conditions, there is no conceptual framework to guide practice and research. On the basis of a review of the literature, this article distinguishes support groups from self-help and treatment groups. It then proposes an open systems model for understanding the factors that affect support groups, for guiding interventions, and for evaluating their outcomes. Four conceptual dimensions are described: (1) environmental conditions (resources and constraints), (2) participant group characteristics (size, composition, and technology), (3) group conditions (goals, structural form, and development), and (4) outcomes (positive and negative effects for members, leaders, and organizations; group problems; and ethical and legal issues). Findings from a pilot study of support group practice based on this framework are presented and discussed. PMID- 8406225 TI - Residential status and the physical health of a mentally ill population. AB - Current literature suggests that severely mentally ill individuals are at high risk for increased physical morbidity and mortality. This study considers the relationship between residential arrangements and the health status of this population. It compares the health status of 234 severely mentally ill individuals living throughout California in sheltered-care facilities, institutions, or the general community. Sheltered-care residence was found to predict positive physical health status when traditional risk factors, as well as risk factors peculiar to this population, were controlled for. The results underscore the value of sheltered-care residence for severely mentally ill individuals who need this type of care. Implications of the results are discussed. PMID- 8406226 TI - Community bioethics: the health decisions community council. AB - Health care decision making in modern society can pose a variety of complex issues for consideration by individuals, families, and providers. The Health Decisions Community Council (HDCC), a community-based bioethics committee, was established to offer a noninstitutional forum for discussion of difficult health care dilemmas. Social work skills and values for autonomy and self-determination are essential to the formation of the HDCC as a model for community bioethics discussions. PMID- 8406227 TI - Enhancing rehabilitation through mutual aid: outreach to people with recent amputations. AB - Social workers in a rehabilitation hospital established an outreach service through which volunteers with previous amputations visited patients with recent amputations, acting as role models and offering emotional support and information. Volunteers received in-depth training and continued with their own system of support. Patients who were visited had strong positive responses to the program. Critical components of the program were developing a collaborative partnership between volunteers and social workers; demonstrating credibility to health care professionals; linking emotional support, information, and affirmation; providing benefits to both patient and volunteer; and strengthening linkages among patient, community, and institution. PMID- 8406228 TI - Health care reform: our greatest opportunity ... ever! PMID- 8406229 TI - "Children, don't forget me": a resource and support group for deployed parents during Operations Desert Shield and Desert Storm. PMID- 8406230 TI - [Physiological functions of endoplasmic and sarcoplasmic reticulum Ca pump and pharmacology of inhibitors of the pump]. AB - This review is derived from the symposium held at the 66th Annual Meeting of the Japanese Pharmacological Society (March, 1993). The symposium consisted of six invited papers whose general theme was the application of recently found ATPase inhibitors selective to SR- and ER-Ca(2+)-ATPase to the analyses of the physiological and pharmacological roles of endoplasmic and sarcoplasmic reticulum Ca stores. Inhibitors used were: thapsigargin, cyclopiazonic acid, 2,5-di-(t butyl)-1,4-benzohydroquinone and 3',3",5',5"-tetraiodosulfophthalein. Gingerol was found to facilitate the action of the ATPase. In either smooth, cardiac or skeletal muscle, sympathetic neurons or several cell lines these inhibitors affected a variety of cell functions and conditions such as contraction, ionic conductance and excitability of the plasma membrane, regulation of intracellular free Ca2+ concentration, transport of viral glycoprotein to the cell surface. Many of these studies utilized either single or cultured cell preparations or skinned muscle. These inhibitors were shown to be useful tools for investigating the SR and ER functioning as Ca sources or Ca sequestrating pumps, and further for estimating the contribution of ER or SR to regulating the flux of Ca2+ and other ions through the plasma membrane. Results of analyses using these inhibitors are discussed. PMID- 8406231 TI - [New trends in studying the regulatory mechanism of smooth muscle contraction]. AB - In this paper, we briefly review current topics about smooth muscle with regards to Ca2+ release, Ca2+ sensitization, and Ca2+ regulation of contraction. Inositol 1,4,5-trisphosphate releases Ca2+ from the sarcoplasmic reticulum, where Ca(2+) dependent immediate feedback control may work. However, the involvement of this feedback control in the Ca(2+)-induced Ca2+ release mechanism remains to be elucidated. Either agonist or GTP gamma S is known to increase the Ca2+ sensitivity of myofilaments. The agonist-induced Ca2+ sensitization could be explained by the up-regulation due to myosin light chain kinase or by the down regulation due to myosin light chain phosphatase. The GTP gamma S-induced Ca2+ sensitization seems to be mediated by rho A p21, a small G protein. Thus, myosin phosphorylation is not the obligatory way to regulate the actin-myosin interaction. We propose that cross-linking between actin and myosin may work as an alternative way to regulate the interaction from biochemical studies. The candidates for the cross-linkers are caldesmon, calponin and myosin light chain kinase. The inhibitory effect of Ca2+ on the interaction, which is observed under the specific conditions for measuring smooth muscle contraction, may hold the key to finding the physiological significance of the cross-linking activity. PMID- 8406232 TI - [New animal models for drug discovery research: focus on cardiovascular diseases]. AB - Development of novel drugs relies on research to discover new drugs. Testing and evaluating new drugs on human subjects are usually impossible because of ethical concerns. Therefore, for drug discovery research, it is essential to establish animal models of human diseases. Numerous animal models have been developed and used in drug discovery and evaluation studies. Such animal models have had important roles in developing new drugs as well as understanding the etiologies of diseases. In the field of cardiovascular drugs, several interesting animal models are currently in use. These include, transgenic mice carrying both human renin and human angiotensinogen genes, Watanabe's heritable hyperlipidemic rabbits, rats with pulmonary hypertension induced by monocrotaline, myocardial infarction model, cardial hypertrophy model and photochemical thrombosis models. It is envisaged that for drug discovery and development, the search for more physiological animal models will continue in the future. PMID- 8406233 TI - [New evolvement of antiarrhythmic drugs]. AB - The Cardiac Arrhythmia Suppression Trial (CAST) casted serious doubts on the usefulness of Class I antiarrhythmic drugs, causing us to turn our attention from drugs impairing conduction of excitation in the myocardium by blocking sodium channels toward those producing increased refractoriness of myocardial cells by blocking potassium channels. This change in the direction of thinking from the "Na+ channel paradigm" to "K+ channel paradigm" resulted in the generation of newly synthetized Class III drugs that have the common electrophysiological property of suppressing outward K+ currents (IK, IK1, Ito) without affecting inward currents (INa, ICa). However, their reversed use-dependence of action potential duration prolonging effect contributes to their untoward action of proarrhythmias. It is still controversial if ion channel blockers acting solely on a certain kind of ion channels are more beneficial than drugs having compound actions such as amiodarone or sotalol. Molecular biology, if combined with arrhythmology, is expected to provide new chemical and pharmacological bases for creating novel antiarrhythmic drugs. PMID- 8406234 TI - [The state of the art of gastric antisecretory drugs]. AB - This review article describes the effect and mechanism of various antisecretory drugs. Both histamine H2-receptor antagonists and gastric proton pump inhibitors are now world-widely used as the first choice for the treatment of acid-related diseases. Because of their potential effectiveness in inhibiting gastric acid secretion, new H2-antagonists and pump inhibitors are under development with the aim of mitigating of the side effects. The local antisecretory effects of FPL 52694, NC-1300-O-3 and ME 3407, which were demonstrated in Heidenhain pouch dogs, will provide clues for the development of new types of antisecretory drugs. Antigastrin drugs are also promising. PMID- 8406235 TI - [Role of drug metabolism studies in the development of new drugs]. AB - The efficacy and toxicity of drugs are closely related to the kinetics and metabolism of drugs. The metabolism of drugs is variable depending on individual, age, sex and species differences. Therefore, in this paper, the following strategies of drug metabolism studies on each phase of development of new drugs is discussed: 1. Early selection of drug candidates for the development of new drugs by a preliminary drug metabolism study. 2. Selection of a drug candidate for the repeated administration of drug toxicity study. 3. Drug metabolism studies on sex- and species-related differences in experimental animals. 4. Drug toxicity and metabolic activation of drug candidates. 5. In vitro and in vivo drug metabolism studies in humans. 6. Arrangement of drug metabolism and pharmacokinetic profile of drug candidate for the phase I study. 7. Role of pharmacokinetics and drug metabolism studies in the phase I study. 8. Pharmacokinetic and metabolic studies for the development of chiral drugs. 9. Approach to drug design from the viewpoint of drug metabolism studies. In conclusion, performance of drug metabolism studies starting from the early stage of drug development is a powerful and useful tool to increase the efficiency and to minimize the period and cost for the development of new drugs. PMID- 8406236 TI - Metabolism and pharmacokinetics of deuterium-labelled di-2-(ethylhexyl) adipate (DEHA) in humans. AB - The metabolism and pharmacokinetics of [2H10]di-2-(ethylhexyl) adipate (DEHA) labelled on the ethyl side-chains was determined in six male volunteers. The dose administered was 46 mg [2H10]DEHA given orally. No parent molecule was found in plasma; however, the metabolite [2H5]2-ethylhexanoic acid (EHA) was detected (mean rate of formation: 1.63 +/- 1.19/hr; mean rate of elimination: 0.42 +/- 0.1/hr). Further oxidative metabolism products were detected in urine; the dominant metabolite identified was EHA present as a conjugate and accounted for an average of 8.6% of the administered dose. [2H5]2-Ethyl-5-hydroxyhexanoic acid, [2H5]2-ethylhexanedioic acid, [2H5]2-ethyl-5-keto-hexanoic acid and [2H5]2 ethylhexanol together accounted for a further 3.5% of the dose. The fate of the remainder was not determined. PMID- 8406237 TI - The disposition of acrylic acid in the male Sprague-Dawley rat following oral or topical administration. AB - The disposition of [1-14C] acrylic acid (AA) was characterized in the male Sprague-Dawley rat following oral administration, by gavage in water, at 400 mg/kg and topical application, in acetone, at 501 micrograms/cm2. The oral dose was well absorbed, rapidly and extensively metabolized, and excreted primarily (approx. 80%) as 14CO2 within 24 hr of administration. The rate and extent of 14CO2 evolution from [14C]AA was greater for [1-14CAA] while a significantly lower proportion of the dosed radioactivity remained in the tissue of animals than that reported for [2,3-14C]AA (Winter et al., Drug Metabolism and Disposition 1992, 20, 665). This is consistent with incorporation of AA into a minor beta-oxidation pathway of mitochondrial propionate metabolism by which AA may be metabolized to CO2 or incorporated into cellular constituents. Approximately 5% of the dosed radioactivity was excreted in the urine. The disposition of [1-14C]AA following dermal application was studied using charcoal containing traps attached to the back of the rats to trap volatilized AA from the dosing sites. Following application of 100 microliters AA [4% (v/v) in acetone] to an area of 8.4 cm2 of the skin of a rat (501 micrograms/cm2), the majority (about 73%) of the dose volatilized and was recovered in the charcoal trap. Percutaneous absorption of AA that did not volatilize was rapid and appeared to have the same metabolic fate as AA administered orally with about 75% of the absorbed dose excreted as 14CO2 within 24 hr. PMID- 8406238 TI - Peri- and postnatal developmental toxicity of beta-myrcene in the rat. AB - beta-Myrcene (MYR) and essential oils containing this monoterpene have been widely used as scenting agents in cosmetics, detergents, soaps, and as flavouring additives in food and beverages. Recently, MYR was reported to be an analgesic substance and the active principle of lemongrass tea. Despite the importance of human exposure to MYR, its toxicological profile has not been comprehensively studied. The aim of this study was to provide data on the peri- and postnatal developmental toxicity of this terpene. MYR (0.25, 0.5, 1.0 and 1.5 g/kg) in corn oil was given by gavage to female Wistar rats from day 15 of pregnancy, parturition and throughout the period of lactation up to weaning (postnatal day 21). The progeny were examined at birth and subsequently to weaning. Mortality, weight gain and physical signs of postnatal development (ear unfolding, incisor eruption, fur development and eye opening) were evaluated. When the exposed offspring reached maturity (120 days) their reproductive capacity was assessed. No adverse effects on the offspring were seen with the lowest dose tested, but 0.5 g/kg and higher doses decreased birth weight, increased perinatal mortality and delayed the day of appearance of landmarks of postnatal development. Moreover, fertility was impaired in female offspring exposed to the two highest doses of MYR. From the data presented in this paper the no-observed-adverse effect level for peri- and postnatal developmental toxicity could be set at 0.25 g beta-myrcene/kg body weight. PMID- 8406239 TI - Altered foci of hepatocytes in rats initiated with diethylnitrosamine after prolonged fasting. AB - The influence of fasting on the potential of diethylnitrosamine (DEN) to initiate liver carcinogenesis was tested in a medium-term assay using the development of putative preneoplastic altered foci of hepatocytes (AFH) as the endpoint. Male Wistar rats fasted for 48 hr were given a single ip injection of DEN (200 mg/kg body weight). Partial hepatectomies were carried out at wk 3 and the rats were killed at wk 8. Fasted rats exhibited a small increase in the numbers of AFH with glutathione S-transferase in the placental form and eosinophilic AFH when compared with non-fasted animals. However, after a 6-wk exposure to 0.05% sodium phenobarbital in the diet, there were no differences in the numbers of AFH between fasted and non-fasted animals. Fasting also increased DEN-dependent centrilobular cell necrosis and specifically drug metabolism as indicated in vivo by a decreased time of paralysis of the lower limbs induced by zoxazolamine (40 mg/kg body weight, ip) and by an unaltered sleeping time induced by sodium pentobarbital (40 mg/kg body weight, ip). The results indicate that although fasting during the initiation stage of carcinogenesis increases DEN hepatoxicity, it does not interfere quantitatively with the development of liver preneoplastic lesions. PMID- 8406240 TI - Effects of the naturally occurring alkenylbenzenes eugenol and trans-anethole on drug-metabolizing enzymes in the rat liver. AB - In order to study the effects of trans-anethole and eugenol on drug-metabolizing enzyme activities in vivo, male Wistar rats were treated by gavage with trans anethole (125 or 250 mg/kg body weight) or eugenol (250, 500 or 1000 mg/kg body weight) daily for 10 days. In liver microsomes and cytosol various phase-I and phase-II biotransformation enzyme activities were determined. No effect on total cytochrome P-450 content in liver microsomes from rats treated with eugenol or trans-anethole was observed. Administration of 1000 mg eugenol/kg body weight, but not the lower doses, significantly increased cytochrome P-450-dependent 7 ethoxy-resorufin O-deethylation (EROD) and 7-pentoxyresorufin O-depentylation (PROD); administration of trans-anethole (125 or 250 mg/kg body weight) did not alter EROD and PROD activities. In rat liver cytosol, UDP-glucuronyl transferase (GT) activity towards the substrate 4-chlorophenol was significantly increased in all treated rats, and activity towards 4-hydroxybiphenyl as substrate was significantly increased in rats treated with 250 mg trans-anethole/kg or with 500 or 1000 mg eugenol/kg. DT-diaphorase (DTD) activity was only significantly enhanced in the liver cytosol of rats treated with trans-anethole at 250 mg/kg body weight. Enhancement of cytosolic glutathione S-transferase (GST) activity towards 1-chloro-2,4-dinitrobenzene was found for all eugenol- and trans-anethole treated rats. In addition, significantly increased levels of GST subunit 2 were measured by HPLC in the liver cytosol of rats treated with eugenol (500 or 1000 mg/kg body eight) or trans-anethole (250 mg/kg body weight). It is concluded that both eugenol and trans-anethole preferentially induced phase II biotransformation enzymes in rat liver in vivo. PMID- 8406241 TI - Propylene glycol-induced skeletal muscle excitation. AB - The effects of propylene glycol (PG) on frog nerve-muscle preparations were examined to determine whether it has an effect on neuromuscular transmission and, if so, to elucidate the mode of the action. PG (5%, v/v) increased the twitch tension to over twice the control value. PG at concentrations above 0.2% significantly increased the amplitude of the endplate potential. PG (1%) raised the frequency of the miniature endplate potentials and increased their amplitude. These results show that PG both facilitates transmitter release from the nerve terminals and raises the acetylcholine sensitivity of the muscle endplate. PMID- 8406242 TI - Changes of hepatic lipid and fatty acid profiles in rats administered iron deficient diet and ethanol. AB - This investigation was undertaken to elucidate the effects of iron deficiency and/or ethanol ingestion on lipid metabolism of female rat liver. 40 Wistar rats (about 40 g weight) were fed a normal diet (Fe: 40 ppm) or an iron-deficient diet (Fe: 5 ppm) for 8 wk. Half of the rats in each group were given 10% ethanol in the drinking water for the last 4 wk. In rats fed the iron-deficient diet (D), the content of total hepatic lipid was higher than that in the rats given the normal diet (N). From the gas chromatography analyses of the fatty acids in total lipids, only the proportion of 18:2n-6 was increased, whereas those of 16:1n-7 and 20:4n-6 decreased. In rats given ethanol and an iron-sufficient diet (NE), the contents of all lipids with the exception of phospholipid were significantly higher than those in the N group. The percentages of fatty acids 12:0, 14:0, 16:0, 18:1 and 20:1 in the total hepatic lipid were increased, whereas those fatty acids of 18:0, 20:4, 22:6 and 24:0 were decreased. In the rats given ethanol and an iron-deficient diet (DE), the contents of all hepatic lipid components did not differ significantly from those in the D group. The percentages of fatty acids 12:0, 14:0 and 16:1 in the total hepatic lipid were increased. These data suggest that ethanol ingestion by iron-deficient rats does not induce the same changes in their hepatic lipid components and fatty acid patterns as those seen in fatty degeneration of the liver. PMID- 8406243 TI - Cooking procedures and food mutagens: a literature review. AB - Commonly eaten meat products prepared from beef, pork, mutton and chicken show some level of mutagenic activity following normal frying. Food preparation methods have a significant influence on the formation of the mutagenic activity. The main food mutagens found in cooked meat products are heterocyclic amines. Several of them have been tested in long-term animal studies and shown to be carcinogenic in rodents. From a health point of view, it is desirable to reduce or prevent the formation of food mutagens. Therefore, a deeper understanding of the precursors and reaction conditions for mutagen formation during normal domestic cooking is very important. Modelling experiments are useful tools for studying the influence of different physical parameters and various precursors on the mutagenic activity. The identification of several thermic mutagens from the modelling experiments support the theory that creatine or creatinine, amino acids and sugars are precursors in the formation of thermic mutagens. Creatine is generally accepted to be a precursor of the mutagens and, interestingly, the conversion of creatine to creatinine has been shown to be blocked by an excess of sugars, which also caused the mutagenic activity to decrease. The mutagenic activity differed for different amino acids used in the model systems, and various thermic mutagens were produced from the amino acids. The incorporation of carbon atoms originating from glucose into food mutagen molecules has shown glucose to be a precursor. Sugar has also been shown to either enhance or inhibit the yield of mutagenic activity, depending on its molar ratio versus the other reactants, which suggests that the Maillard reaction may be used to control the formation of mutagens. PMID- 8406244 TI - Aluminium and Alzheimer's disease--an update. PMID- 8406245 TI - Second metatarsophalangeal joint instability in the athlete. AB - In a group of athletically active patients, second metatarsophalangeal joint instability was diagnosed in nine patients (11 toes). A positive drawer sign was pathognomonic of early second metatarsophalangeal joint instability. A soft tissue realignment procedure was used to stabilize the second metatarsophalangeal joint in seven toes. In five of seven cases (71%), good to excellent results were noted at an average follow-up of 20.4 months. PMID- 8406246 TI - Comparative study of therapies for fibular ligament rupture of the lateral ankle joint in competitive basketball players. AB - This retrospective study compares the results of different therapies for fibular ligament rupture in a homogenous group of professional athletes. The endpoint "competitive sports" was an outcome consideration. Subjects were examined by means of a standardized questionnaire and a structured interview. One hundred and seventy-nine of the questionnaires were completed and returned for evaluation. All of the basketball players with severe ankle sprain (supination trauma with swelling, pain, and inability to bear stress) were included. Those players with fractures of the foot, pronation trauma, or additional distal fibula or tibia fractures were excluded from this study. Of the 179 basketball players 160 (89%) had suffered severe ankle sprain. The treatment was divided into three groups: primary surgery (N = 35), plaster cast (N = 39), and functional treatment (N = 89). While simple ligament injuries (Grade I and II) were mostly treated functionally, complex ligament injuries (Grade III) were usually operated on. A total of 119 (74%) of the players reported no further pain. For pain reduction surgical and functional treatments showed advantages over plaster treatment. In the surgical group 63% of the players judged their regained stability to be equivalent to that of their healthy leg. Only 50% of the players in the plaster and functional groups believed their ankle joints to have regained the same stability as before their injuries. Despite the achievement of good results through surgery, there were clear differences in the players' assessments of their performance in competitive sports. Most subjects (92%) did not have any problems in everyday life regardless of which kind of therapy had been chosen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406247 TI - Surgical treatment for mild deformities of the rheumatoid forefoot by partial phalangectomy and syndactylization. AB - Thirteen patients (14 feet) were treated for mild rheumatoid forefoot deformities with lesser toe partial proximal phalangectomies and partial syndactylizations. Eleven patients (85%) were reviewed at an average of 8 years postoperatively. The results were completely satisfactory in four patients, satisfactory with minor reservations in three patients, satisfactory with major reservations in one patient, and unsatisfactory in three patients. The major cause of reservations and lack of satisfaction was metatarsalgia. Seven patients (64%) reported that their activities were limited by intermittent metatarsalgia. Four patients (36%) considered the cosmetic appearance of the forefoot to be unsatisfactory. All but one patient required some form of shoewear modification. Based on this study, we believe the indications for this procedure are limited. These include rheumatoid patients with mild forefoot deformities without significant metatarsalgia or ongoing disease who have failed nonoperative treatment. Relative contraindications to this operation appear to include the recent onset of rheumatoid arthritis, active disease, significant metatarsalgia, and strong cosmetic concerns regarding outcome. In borderline clinical decisions that involve whether or not to leave or excise the lesser metatarsal heads, they probably should be excised to decrease late metatarsalgia. PMID- 8406248 TI - Lateral ankle instability as a cause of superior peroneal retinacular laxity: an anatomic and biomechanical study of cadaveric feet. AB - The role of the competent superior peroneal retinaculum (SPR) as a primary restraint to peroneal tendon subluxation and mechanical attritional wear is clear. Injury to the SPR has classically been described as a dorsiflexion eversion movement of the ankle coupled with a forceful peroneal tendon reflex contraction. This mechanism, however, does not cause injury to the lateral collateral ligaments of the ankle and does not explain the coexistent findings of lateral ankle instability, laxity of the SPR, and concurrent peroneal tendon pathology. Anatomic studies reveal a parallel alignment of the calcaneal band of the SPR and the calcaneofibular ligament. A cadaveric model of ankle instability created by serial sectioning of the lateral collateral ligaments revealed increasing visual strain on the SPR with increasing degrees of ankle instability. These findings suggest the SPR serves as a secondary restraint to ankle inversion stress and that the force or forces that result in chronic ankle instability can also injure and attenuate the superior peroneal retinaculum. PMID- 8406249 TI - SPLATT combined with tendo achilles lengthening for spastic equinovarus in adults: results and predictors of surgical outcome. AB - During a 4-year period, split anterior tibial tendon transfer (SPLATT) was performed on 42 adults with cerebrospastic equinovarus deformity. Twenty-one patients (24 feet) had a minimum 1-year follow-up, which included detailed documentation of foot appearance position and function as well as ambulatory status. Thirteen patients were male and 8 were female. Average age of the patients was 41 years. Seventeen patients were independent ambulators with orthoses, one was a maximally assisted ambulator. Three patients with spastic quadriparesis were nonambulatory. All patients had uniform surgical technique and postoperative management. This paper presents the results of SPLATT and identifies risk factors for poor surgical outcomes. After an average follow-up of 39 months, 83% of the feet were rated as having good or excellent results. All ambulatory patients had improved gait and 35% of them were able to discontinue their orthoses. Poor surgical outcomes were associated with nonambulatory status in brain injured patients (P = .018). Salvage of failed SPLATT is discussed. PMID- 8406250 TI - Extensor hallucis longus coaptation to tibialis anterior: a treatment for paralytic drop foot. AB - Between June 1982 and April 1983, a procedure to coapt the extensor hallucis longus (EHL) to the tibialis anterior was performed in eight post-polio patients to correct drop foot and to enable the EHL to be a more efficient dorsiflexor of ankle. Although at early follow-up, every patient was able to actively dorsiflex the ankle against gravity, at final review, (mean follow-up 7.8 years), only two patients could still do so. Three patients developed a cock-up toe deformity or dorsiflexion deformity of great toe. We have attributed the poor final results to stretching of the coaptation. Use of splints or orthosis for a longer period postoperatively and a more carefully designed physical therapy may have yielded better results. Alternatively, if the EHL is anchored to navicular bone better results may be obtained. PMID- 8406251 TI - Protective sensation of the plantar aspect of the foot. AB - The scientific literature suggests that barefoot activity may be beneficial. There is a current trend in recreational barefoot activity in children and adults, and barefoot running among athletes. Although the type of skin over most of the body (hairy skin) seems to be easily injured by painful abrading loads, little is known about protection provided by plantar sensory feedback against damage from excessive wear during barefoot locomotion. To evaluate this, we administered a volley of 35 painful abrading loads to glabrous and hairy skin sites over a 5-min period, and examined its effects for signs of cutaneous injury in a sample of 12 normally shod healthy male subjects. Compared with hairy skin of the thigh, plantar skin required approximately 600% greater abrading loads to reach pain threshold. Furthermore, painful stimulation produced visible redness and hypersensitivity in all subjects at the hairy skin site 24 hr after stimulation, whereas only 8.3% reported hypersensitivity and none showed erythema at the plantar area 1 day later. We found that plantar skin possesses a higher pain threshold to abrading stimuli than hairy skin. In fact, loading of the plantar area was limited to innocuous levels due to intolerable pain. We conclude that plantar skin is well protected through sensory feedback from abrasive injuries when barefoot. This information combined with previous reports suggests that risk of injury when normally shod individuals perform barefoot locomotion should be low. PMID- 8406252 TI - Biomechanical evaluation of longitudinal arch stability. AB - In spite of the common occurrence of pes planus and multiple operations that have been reported to relieve the associated symptoms, there is little published on the relative contribution of various structures to stabilization of the arch of the foot. Twelve fresh-frozen human cadaveric feet were loaded along the tibial axis with compressive loads of 230, 460, and 690 newtons with the specimens intact and after sequential sectioning of plantar fascia, plantar ligaments, and spring ligament. Structures were sectioned in six different sequences and changes in vertical and horizontal dimensions of the medial arch were measured. The highest relative contribution to arch stability was provided by the plantar fascia, followed by plantar ligaments and spring ligament. Plantar fascia was a major factor in maintenance of the medial longitudinal arch. Its division in the cadaveric feet decreased arch stiffness by 25%. PMID- 8406253 TI - Jones' fractures and related fractures of the proximal fifth metatarsal. AB - At least three fracture types occur in the proximal fifth metatarsal: the Jones' fracture, the proximal diaphysial stress fracture, and the tuberosity avulsion fracture. Each has distinct characteristics. The diaphysial stress fracture is commonly confused with the Jones' fracture, thereby obscuring vital differences in prognosis and treatment. Anatomical and biomechanical characteristics, as well as vascular studies of the proximal portion of the fifth metatarsal, are discussed in an attempt to better understand their diverse healing potentials. Guidelines for treatment are controversial, and must frequently be individualized. Although surgical intervention for certain proximal fifth metatarsal fracture types may speed recovery time, most fractures heal with immobilization. Treatment of displaced, intra-articular fractures, delayed unions, and nonunions usually requires operative methods. PMID- 8406254 TI - Hypertrophy of the abductor hallicus muscle: an unusual congenital foot mass. AB - A rare case of an isolated congenital muscle hypertrophy involving the abductor hallicus muscle is presented. Although the entity has not been well described previously, it should be entertained in the different diagnoses of medial childhood foot masses. Presently, the diagnoses of such masses should be made more easily with the widespread availability of magnetic resonance imaging. Treatment is generally straightforward, consisting of observation alone, versus partial or total excision, as dictated by the operative findings and previous functional impairment. PMID- 8406255 TI - Treatment of infected pilon nonunions with small pin fixators. AB - Six consecutive patients with infected intra-articular fractures of the distal tibia were studied. They were treated with resection of all infected or necrotic bone, systemic antibiotics, and instrumentation with a small pin fixator (Ilizarov external fixator). The fixator was used to perform an ankle arthrodesis and to fill the defect created by bone resection with distraction osteogenesis. In all cases, the infections were eradicated, and a solid arthrodesis was attained. The patients required from zero to two revision procedures (average 1.3), and their time in the fixator varied from 3 to 13 months (average 8 months). All of the patients experienced at least minor complications during treatment (superficial pin tract infections). At final follow-up, no patient demonstrated shortening of more than 1.5 cm. One patient has an internal rotation deformity of 15 degrees; a second has a varus deformity of 10 degrees and occasionally uses lateral support (a cane) secondary to unsteadiness on uneven ground; and one patient uses aspirin occasionally for subtalar pain. All are pleased with their results and would undergo the same procedure again without reservation. PMID- 8406256 TI - Comminuted fracture of the calcaneus associated with subluxation of the talus. AB - Two cases of complex fracture dislocation of the calcaneus having an unusual pattern of injury are described. The cases exhibit the following special characteristics: (1) fracture dislocation of the calcaneus where the primary fracture line separates the calcaneus into an anteromedial fragment that maintains its normal relationship to the talus and a posterolateral fragment that is dislocated from the subtalar joint. This posterolateral fragment moves laterally and lies adjacent to the fibula; (2) a secondary fracture line separating the lateral portion of the posterior facet from the tuberosity of the calcaneus. Both fragments are dislocated from their normal anatomical position; (3) talar tilt as shown on AP view of the ankle caused by inversion of the talus due to rupture of the lateral collateral ligament. Also, the posterolateral fragments impinging on the fibula pushes the heel downward and contributes to the talar tilt; (4) involvement of the calcaneocuboid joint; (5) dislocation of the peroneal tendons. This fracture pattern is unusual and has not been described before. Recognition of this unusual injury with subsequent and proper management may prevent major disability to the patient. Conservative treatment by casting or early range of motion is contraindicated. Closed reduction should be attempted immediately, and if not successful, a lateral approach with open reduction and internal fixation is the treatment of choice for this complex injury. PMID- 8406257 TI - Flexor tendon transfer for metatarsophalangeal instability of the second toe. AB - Flexor to extensor transfer was used to treat painful second metatarsophalangeal joint instability in thirteen feet in eleven patients. All patients had their pain reproduced with vertical stress motion of 50% to 100% at the metatarsophalangeal joint. Seven feet had concomitant hallux valgus correction, two feet had no hallux valgus, and four feet underwent no correction for asymptomatic hallux valgus. Results at an average of 33.4 months followup showed that all patients had substantial pain relief, with eight patients becoming pain free, and five patients experiencing mild pain. All but one were satisfied with their result. Stiffness appeared to be the source of the mild residual pain. All toes, including six toes with preoperative medial crossover toe deformity, were corrected into valgus alignment with adjacent toes. All toes operated on for the first time were able to touch the ground with grasp postoperatively. Flexor to extensor transfer is successful in reducing the second toe and relieving pain caused by instability of the second metatarsophalangeal joint, but may require rapid postoperative mobilization to ensure passive dorsiflexion equal to that of the adjacent toes to reduce postoperative uncomfortable stiffness. PMID- 8406258 TI - Fatty acid composition of normal and atrophied heel fat pad. AB - Capillary gas-liquid chromatography was used to analyze the fatty acid composition of normal heel fat pads from subjects without systemic disease (N = 8) and atrophied heels from patients with diabetic peripheral neuropathy (N = 4), rheumatoid arthritis (N = 1), peripheral vascular disease (N = 1), and hereditary sensory neuropathy (N = 1). In the normal subjects, the fatty acid composition of subcutaneous abdominal fat was also obtained for comparison. Three saturated fatty acids (myristate, palmitate, and stearate) and four unsaturated fatty acids (palmitoleate, oleate, vaccenate, and linoleate) comprised over 90% of the total fatty acid composition. Higher percentages of unsaturated fatty acids and lower percentages of saturated fatty acids were found in the normal heel fat pads when compared to subcutaneous abdominal fat. The increase in the ratio of unsaturated fatty acids to saturated fatty acids (4.4 versus 2.5, P < .01) may decrease triglyceride viscosity and enhance the biomechanical efficiency of the heel fat pad. Though the number of patients is small, no statistically significant compositional differences were noted between the heel fat from normal subjects and from subjects with peripheral neuropathies, rheumatoid arthritis, or peripheral vascular disease. However, the heel fatty acid composition of the one subject with a hereditary sensory neuropathy was less unsaturated and more saturated than normal with a ratio of unsaturates to saturates similar to that of the abdomen (2.8). PMID- 8406259 TI - Arthrodesis of the first metatarsophalangeal joint: a biomechanical study of internal fixation techniques. AB - This study compares the strength and rigidity of four methods of internal fixation for arthrodesis of the first metatarsophalangeal joint. Ten matched pairs of cadaveric first rays were harvested and arthrodesis performed by one of four techniques: (1) planar excision of joint surfaces and fixation with crossed Kirschner wires, (2) planar excision of joint surfaces and internal fixation with a dorsal plate and screws, (3) planar excision of joint surfaces and internal fixation with an interfragmentary screw, or (4) excision of the joint surfaces using powered conical reamers and fixation with an interfragmentary lag screw. Biomechanical testing with a Bionix 858 materials testing machine was carried out, applying a plantar force utilizing principles of cantilever loading. Force applied and displacement of the arthrodesis were recorded. Of the four methods tested, bony preparation with power conical reamers and supplementary interfragmentary screw fixation was the most stable. PMID- 8406260 TI - Achilles tendon injuries: a comparison of surgical repair versus no repair in a rat model. AB - Controversy exists regarding the treatment of Achilles tendon ruptures. The aim of this study was to determine whether surgical repair of the rat Achilles tendon offered any biomechanical, functional, or morphological advantage over no repair. Thirty-two male Sprague-Dawley rats were randomly allocated into four groups: (1) sham operated (skin incision only), (2) no repair (complete division of the Achilles tendon and plantaris tendon without repair), (3) internal splint (plantaris left intact), and (4) Achilles repair (with a modified Kessler-type suture). Functional performance was determined from the measurements of hindpaw prints utilizing the Achilles Functional Index. On day 15, the animals were killed, and biochemical and histological evaluations were performed on both the injured and uninjured Achilles tendon constructs. All groups subjected to Achilles tendon division had a significant initial functional impairment that gradually improved so that by day 15 there were no functional or failure load impairments in any group. The injured tendons in all three groups subjected to Achilles tendon division had a 13-fold increase in the cross-sectional area and were less stiff and more deformable than uninjured and sham-operated tendons on day 15 (P < .001). The magnitude of the biomechanical and morphological changes at postoperative day 15 and the initial impairment and rate of functional recovery were similar for no repair, internal splint, and Achilles repair groups. In summary, this study demonstrates that surgical repair of the Achilles tendon in the rat does not offer any advantage over nonoperative management. PMID- 8406261 TI - Tarsal tunnel syndrome secondary to cosmetic silicone injections. AB - Silicone has been implicated as a cause of inflammatory disorders in the body including synovitis and lymphangitis. Silicone particulate matter has been shown to cause a fairly severe chronic foreign body reaction with the use of silicone prosthesis in the foot. This reaction was often overlooked for years because of the limited number of subjective complaints. There are case reports of granulomatous inguinal lymphadenopathy following first metatarsophalangeal joint silicone arthroplasty. Similar findings have been noted in the axilla with hand implants and breast implants. Of greater significance are two reports of malignant lymphoma found with silicate lymphadenopathy. PMID- 8406262 TI - Calcaneal fracture-dislocation with entrapment of the medial neurovascular bundle: a case report. AB - The calcaneus is the most commonly fractured of all the tarsal bones. Soft-tissue interposition, usually involving the peroneal tendons, has been reported to block reduction of calcaneal fractures and subtalar dislocations. To our knowledge, no case of entrapment of the neurovascular bundle has been reported. This is the subject of our report. PMID- 8406263 TI - Spontaneous atraumatic rupture of the flexor hallucis longus tendon under the sustentaculum tali: case report, review of the literature, and treatment options. AB - A case of spontaneous rupture of the flexor hallucis longus tendon within the hind part of the foot is reported in a middle-aged woman who had no trauma or systemic disease. Repair was effected by tenodesis to the flexor digitorum longus tendon above and below the fibro-osseous tunnel. Hyperextension of the interphalangeal joint which most troubled the patient was corrected postoperatively. Active range of motion was equal in extension. Flexion at the IP joint was present, but significantly less than the unaffected side. PMID- 8406264 TI - The Rochester bone trephine for small joint arthrodesis in the foot. AB - The Rochester bone trephine is a useful instrument for performing small joint arthrodesis in the foot. The technique involves harvesting a dowel graft from the iliac crest and then inserting the graft into a recipient bed across the small joints in the foot. Minimal surgical trauma and patient discomfort result from this technique. Three cases are presented to illustrate the usefulness and diversity of this system. PMID- 8406265 TI - Speech motor control in Parkinson's disease: a comparison between a clinical assessment protocol and a quantitative analysis of mandibular movements. AB - Nine individuals with Parkinson's disease (PD) and nine normal control subjects, matched for sex and age, were compared on a standardized assessment protocol for dysarthria. Simple measures of hand and gait motility were also obtained. A kinematic analysis of vertical jaw movements during speech was performed by using an optoelectronic technique. The PD subjects were examined during both induced off L-dopa and on L-dopa states, and their performance was compared between the two conditions. Significant group differences were found for all kinematic measures during a syllable repetition task, as well as for the total dysarthria test scores and certain individual test items. Implications and clinical usefulness of the kinematic approach to the study of oral motor functions are discussed. PMID- 8406266 TI - Kinematics of the prominentia laryngea at phonation onset. AB - The present paper deals with the problem of the temporal relations between the beginning of the glottal opening at phonation onset and the thyroid cartilage kinematics at the prominentia laryngea. Furthermore, some differences in comparison with a stationary phonation event are discussed here. PMID- 8406267 TI - Acoustic pattern recognition of fricative-vowel coarticulation by the self organizing map. AB - Word-initial samples of fricative [s] preceding vowels [a:], [ae:], [e:], [i:], [u:], [o:], and [y:] in Finnish words were studied with the self-organizing map. An acoustic map was first calculated from speech samples of women without speech disorders, and then the [s] samples were measured on this map. In all 10 subjects the [s] samples preceding the rounded vowels [u:] and [o:] clearly differed from the samples in front of unrounded [a:], [ae:], [e:], and [i:]. The coarticulatory phenomena observed on the map were due to changes in the composition and stability of the fricative spectrum. PMID- 8406268 TI - [Evaluation of laryngeal sound generation with FFT analysis of glottic impedance in patients with recurrent nerve paralysis]. AB - The electroglottogram (EEG) is known to be related to the motion of vocal folds. The major hypothesis is that the EGG is related to the area of contact of the vocal folds. EGG is thought to be incapable of measuring or monitoring non contact type vibratory events in the larynx. Recently it was demonstrated by fast Fourier transformation (FFT) analysis of EGG curves that the resulting EGG can be interpreted as a sum of a 'laryngeal fundamental with overtones'. This necessitates a new interpretation of normal EGG waveshape. This study was undertaken to prove the hypothesis that (pathological) EGG curves contain information even about non-contact type vibratory events in the larynx during phonation when analyzed by FFT. Patients with vocal fold paralysis were asked to phonate sustained vowels (/a/, /e/, /i/, /o/, /u/). Voice sounds and EGG signals were recorded simultaneously, routed to a frequency analyzer and subjected to real-time FFT analysis. In all cases original EGG signals would have been classified as pathological. However, in all recordings the laryngeal fundamental frequency and--in some patients--overtones with their intensities could be extracted and compared to harmonics extracted from voice sound. This is the first report of FFT analysis of EGG signals in patients with vocal fold paralysis. It is proposed that vocal folds vibratory events may be of non-contact type and that they can be monitored by analysis of glottic impedance. PMID- 8406269 TI - Emotions in the sight of long-time averaged spectrum and three-dimensional analysis of periodicity. AB - The authors used long-time averaged spectrum and three-dimensional analysis of periodicity (3D-PAN) for analyzing emotional expressions in utterances. They confirmed the movement of the skewness of formant region, that is getting smaller at anger and at joy. It can reach positive values. The skewness is higher at sadness. These findings correspond to the physiological phenomena that we can follow in the whole musculature. The 3D-PAN has not brought any new finding as far. PMID- 8406270 TI - [Therapy of iron deficiency. Diagnostic prerequisites--therapeutic procedures]. AB - Iron is an essential trace element, and iron deficiency is common. Underlying causes must be carefully investigated (occult bleeding, carcinoma). Mild forms of iron deficiency may be due to rapid growth, an inappropriate diet, menstrual bleeding or pregnancy. As a rule, they can be corrected by dietary means. Pharmacological doses of iron may irritate the intestinal mucosa, limiting the size of the individual dose and impairing compliance. Overdosing can be fatal. The duration of medication depends upon the rapidity with which deficient hemoglobin iron and iron stores can be replenished. Failure of treatment may be due to a wrong diagnosis, persistent bleeding or disturbed absorption. Parenteral administration of iron should be reserved for patients in whom oral iron replacement fails. PMID- 8406271 TI - [Irrational fear of cortisone. A study of attitude and information status of mothers]. AB - METHOD: 200 mothers-with-child sent to a health resort were asked to complete a questionnaire and submitted to an interview to determine their attitudes toward the use of corticosteroids. RESULTS: A total of 178 proved to be fairly well informed. Of these, 123 had reservations. However, only a small number refused to countenance such treatment at all. Seventy-two mothers and 45 of their children had had personal experience with corticosteroids. Obvious deficiencies in the quality of information were detected, in particular with respect to the differing risks of short- and long-term treatment. A desire to receive adequate information from the care-providing physician was expressed, while package inserts were generally viewed with critical skepticism. PMID- 8406272 TI - [Dyspepsia and acid]. PMID- 8406273 TI - [Weight reduction with alternative diets. Principles, philosophy, side effects, uses. Part 4: Alternative diets (H)]. PMID- 8406274 TI - [Mofebutazone--evaluation of possible interaction with glibenclamide]. AB - METHODS: In an open clinical trial involving 20 patients, the question was investigated as to whether the administration of 3 x 300 mg mofebutazone daily resulted in any clinically relevant changes in blood sugar levels in diabetics treated with glibenclamide. RESULTS: Fluctuations observed, both in mean and individual values, were within the range of those regularly seen in diabetics. This was shown by both weekly measurements and a comparison of daily profiles. A comparison of the daily blood sugar profile prior to and after three weeks of mofebutazone, revealed the following mean figures (+/- STD): prior to mofebutazone 12.30 hours 155.5 (+/- 23.2) mg/dl, 16.30 hours 147.2 (+/- 22.0) mg/dl; after 3 weeks of mofebutazone: 12.30 hours 162.7 (+/- 20.0) mg/dl, and 16.30 hours 151.7 (+/- 17.7) mg/dl. No side effects occurred, and no patient left the study prematurely. The laboratory parameters also studied showed no remarkable changes, either in mean values or in individual figures. PMID- 8406275 TI - [HIV infection transmissible also by leeches]. PMID- 8406276 TI - [Alternative procedures: not only of medical history interest. Constitutional therapy of Berhard Ascher]. PMID- 8406277 TI - [Current status of cementless hip endoprostheses ]. AB - In young patients, the results of cemented hip joint replacement are less than satisfactory on account of the high rate of aseptic loosening involving in particular the acetabular component. With the aid of suitable uncemented prostheses and a meticulous surgical technique, the results obtained in this group can be improved. On an interim basis, an uncemented acetabular component may be combined with a cemented femoral component. The problem of wear and foreign body-induced bone resorption remains unsolved. New materials will possibly rectify this problem. Customized prostheses have opened up new possibilities, in particular for the severely deformed hip joint, but whether they will improve the results of standard replacements has yet to be shown. Before introducing a new prosthesis, thorough use should be made of the testing possibilities available. In the elderly patient with a reduced life expectancy, the cemented prostheses continue to be the method of first choice, since it permits immediate pain-free weight-bearing. PMID- 8406278 TI - [Special endoprostheses of the knee joint in bone tumors]. AB - Since the introduction of more effective adjuvant therapies in the area of primary and secondary bone tumors, limb salvage procedures have become more and more important. At the Orthopedic Department of the Technical University, Munich, 52 special knee prostheses have been implanted in patients with malignant tumors of the distal femur and proximal tibia. On the basis of the ISOLS criteria, the functional results achieved were excellent in 14%, good in 47%, satisfactory in 35%, and poor in only 5% of patients after an average follow-up period of 43 months (range: 3-156 months). The perioperative complication rate was 27%, and late complications were material fatigue and wear. In the case of primary tumors, a customized prostheses, and in the case of secondary tumors a modular system is recommended. Individual treatment planning is essential. PMID- 8406279 TI - [Incidence of Lyme borreliosis in a rheumatologic patient sample. Study of 153 patients of an internal medicine-rheumatologic ambulatory clinic]. AB - PROBLEM: Arthritis of the large joints is considered a typical manifestation of Lyme disease. With an estimated incidence in the population of about 2%, Lyme disease arthritis would be expected to be a fairly common diagnosis in a rheumatology unit. METHODS: In 153 successive patients seen in a rheumatology clinic of the University Hospital Frankfurt am Main, who had historical or clinical evidence of arthritis, the IgG antibody titers against Borrelia burgdorferi in the serum were compared with the clinical presentation. RESULTS: Twelve patients had positive or borderline positive antibodies titers. In three of the 153 patients, the diagnosis of Lyme disease-associated arthritis appears possible, although not probable. CONCLUSIONS: At least in the greater area of Frankfurt, Lyme disease as a differential diagnosis of arthritis is apparently of no great significance. PMID- 8406280 TI - [Weight reduction with alternative diets. 5: Alternative diets (I)]. PMID- 8406281 TI - [Is early combined AIDS treatment an advantage? Nucleoside analogs--study will bring enlightenment]. PMID- 8406282 TI - [RU 486: safe and wrongfully maligned]. PMID- 8406283 TI - [Perception of and psychological coping with medical technology. How does the patient perceive percutaneous transluminal coronary angioplasty?]. AB - BASIC REMARKS: Since, as a result of technological progress, ever more interventions are being carried out in the conscious patient without anesthesia or sedation, psychological coping is an important aspect. METHODS: Over a period of one year, all patients undergoing coronary artery dilatation at the University Hospital, Frankfurt, were admitted to the study in which the patients and their spouses, as well as the physicians involved were questioned (individual questioning, questionnaires, interviews). In addition, a catamnestic follow-up examination was performed. RESULTS: In addition to numerous other results, the following major points can be made: most patients coped with the stressful situation by adopting an optimistic attitude, which was combined with trust in the physician and the procedure employed. A very small group of patients can be considered to suffer from appreciable depressivity/anxiety and requires psychotherapeutic support. Days and weeks after the procedure there is a delayed phase of "late coping". In the patients with no major persisting symptoms, there is an increasing tendency in this period to play down the situation, with the result that there may be a lack of readiness to admit the need to change possibly dangerous habits and adopt a different life style. PMID- 8406284 TI - [Conservative therapy of backache. Part 4: Manual therapy]. AB - The literature on the clinical effectiveness of manipulation in low back pain is extensive, although scientific studies that are free from flaws are much rarer. Most of the studies so far done show--with some reservations--a positive effect for manipulation. Given the proper indication, and with care being taken to observe contraindications, this modality can thus be recommended if performed by an experienced manipulator. However, this should not blind us to the fact that further investigations are necessary, and that the technique has no unequivocal advantages over exercise. PMID- 8406285 TI - [Weight reduction with alternative diets. Principles, philosophy, side effects and uses. Part 6: Alternative diets (K through L)]. PMID- 8406286 TI - [Long-term treatment of hyperlipoproteinemia with etofibrate]. AB - In the treatment of hyperlipoproteinemia, longterm results are of particular interest. Fifty-five patients with mixed hyperlipidemia, who had been treated with Etofibrat in a controlled clinical trial, underwent a follow-up examination one year after conclusion of the study, and data of current lipid treatment and lipid status were recorded. Also investigated were the compliance and motivation of the patients. Within the course of the clinical study, patients with combined hyperlipidemia showed a marked decrease in cholesterol levels of 21%, and of triglycerides of 38%. At the end of the trial, 20 patients continued to receive Etofibrat. Under this treatment, the lowered cholesterol and triglyceride levels were maintained, while in patients receiving other lipid-lowering agents, the triglycerides rose again, and patients not receiving any further medication showed an increase in both triglyceride and cholesterol levels. PMID- 8406287 TI - [Prevention of arteriosclerosis from the very beginning. Therapy with specific synergistic vitamins against hyperhomocysteinemia]. PMID- 8406288 TI - A courier experience. PMID- 8406289 TI - FNS plans for birthing center. PMID- 8406290 TI - Epithelial factors: modulation of the airway smooth muscle tone. AB - The airway epithelium is composed of a heterogeneous population of cells. This epithelial layer is not only a physical barrier but also a target responding to a variety of inflammatory mediators. These cells can respond by releasing contracting and relaxing factors to modulate airway responsiveness. They can also metabolize some of the inflammatory mediators. Epithelial damage is a consistent feature of some respiratory conditions, but whether or not such damage contributes to airway disease is for the moment unknown. This review summarizes the literature on the known and proposed roles of the epithelium in the modulation of the airway smooth muscle tone. PMID- 8406291 TI - A study of alpha 1- and alpha 2-adrenoceptor responsiveness in diabetic insipidus dogs. AB - The present study was performed to investigate the participation of circulating vasopressin in alpha-adrenoceptor responsiveness. Thus, we compared the pressor responses induced by selective alpha 1-or alpha 2-adrenoceptor stimulation in two groups of conscious dogs: a) normal animals and b) animals with surgically induced diabetes insipidus. In addition, platelet alpha 2-adrenoceptors labelled with (3H)RX821002 were compared in the two groups. The pressor response to alpha 1-adrenoceptor stimulation [ie successive doses of noradrenaline (0.5, 1, 2, 4 micrograms/kg i.v.) after propranolol (1 mg/kg i.v.) plus yohimbine (0.5 mg/kg i.v.)] was significantly (P < 0.05) less pronounced in diabetic insipidus than in normal dogs. In contrast, the magnitude of the pressor effects of alpha 2 adrenoceptor stimulation [ie noradrenaline after propranolol plus prazosin (1 mg/kg i.v.)] was the same in the two groups of animals. Bmax and Kd values for (3H)RX821002 binding on platelets were similar in diabetic insipidus and normal dogs. This study shows that alpha 1- (but not alpha 2-) adrenoceptor responsiveness is decreased in diabetic insipidus suggesting the involvement of vasopressin in the mechanisms of the vascular alpha 1-adrenoceptor pressor response. PMID- 8406292 TI - Bronchopulmonary responses to endothelin-1 in sensitized and challenged guinea pigs: role of cyclooxygenase metabolites and platelet-activating factor. AB - The effect of phosphoramidon on the increase in pulmonary inflation pressure (PIP) induced by endothelin-1 (ET-1) administered by aerosol in ovalbumin (OA) sensitized and challenged guinea-pigs was investigated after pretreatment or not of the animals with the neutral endopeptidase inhibitor, phosphoramidon, the cyclooxygenase inhibitor, indomethacin or the platelet activating factor (PAF) antagonist, BN 50730. When guinea-pigs were pretreated by phosphoramidon (0.1 mM, aerosol for 15 min), a significant enhancement of PIP was observed after administration of ET-1 (1 or 3 micrograms ml-1, aerosol for 2 min), whereas these doses of the peptide were only slightly active in control animals. In sensitized and unchallenged guinea-pigs, ET-1 (1 or 3 micrograms.ml-1, aerosol for 2 min) induced, as in controls, a moderate increase in PIP. In contrast, aerosol exposure of OA in sensitized guinea-pigs developed an increased PIP following ET 1 (1 and 3 micrograms.ml-1, aerosol for 2 min) administration, that was non significantly affected by pretreatment of the animals with phosphoramidon after the dose of 3 micrograms ml-1 ET-1. Guinea-pigs exposed to phorphoramidon and treated with indomethacin (10 mg kg-1, i.v.) or BN 50730 (25 mg kg-1, per os) significantly reduced the increase in PIP upon administration of ET-1 (3 micrograms.ml-1, aerosol for 2 min). No inhibitory effect of indomethacin was noted when ET-1 (3 micrograms.ml-1, aerosol for 2 min) was administered to sensitized and OA-exposed guinea-pigs, pretreated or not with phosphoramidon. In contrast, BN 50730 significantly reduced the increase in PIP induced by ET-1 observed in sensitized and OA-exposed guinea-pigs. Moreover, this drug was moderately active in reducing the increase in PIP induced by ET-1, when the animals were pretreated by phosphoramidon. These results suggest that a phosphoramidon-sensitive endopeptidase-like enzyme, present in the airway tissue modulates the effect of ET-1. Furthermore, the increase in PIP to ET-1 observed in aerosol-sensitized and antigen-exposed guinea-pigs appears to be mediated by PAF rather than cyclooxygenase metabolites, even though the participation of other mediators in this process is open. PMID- 8406293 TI - Physiological roles of dopamine and neuropeptides in the retina. AB - The retina is a highly complex nervous tissue that converts light into patterns of electrical action potentials in order to process visual information. To carry out its function as a transducer and processor of visual information, the retina must be structurally and biochemically organized to send a coherent signal to the visual areas of the brain. In recent years, a number of biologically active substances have been demonstrated to be located within neurons in the retina. Most of them are thought to be involved in the modulation of the signal and its transmission to the brain through the optic nerve. The present paper attempts to summarize the immunocytochemical distribution and physiology of some neuronally localized substances in the mammalian retina, namely dopamine and neuropeptides. PMID- 8406294 TI - Effect of zolpidem during sleep on ventilation and cardiovascular variables in normal subjects. AB - A double-blind study comparing the effect of zolpidem 10 mg, and placebo, on sleep architecture, nocturnal ventilation, cardiovascular parameters (heart rate, systolic, diastolic and mean blood pressures) was carried out. Ten healthy middle aged males took part in the study. No significant differences were found between zolpidem and placebo in relation to sleep architecture. Mean respiratory disturbance index (RDI) and SaO2 values (mean SaO2, time spent with SaO2 < 90%) were similar under both conditions. The diastolic and mean blood pressure readings taken from REM periods which occurred during the first third of the night were significantly higher with zolpidem. No changes in systolic blood pressure or heart rate were found with zolpidem in comparison to placebo. PMID- 8406295 TI - Lack of effect of phenobarbital on ex vivo leukocyte oxidative metabolism in healthy volunteers. AB - Oxidative metabolism in activated human polymorphonuclears catabolizes leukotriene B4 by a cytochrome P450 omega-hydroxylase and procainamide by a myeloperoxidase. The percentage of leukotriene B4 metabolized by activated human polymorphonuclears and the apparent enzymatic parameters of procainamide metabolism were studied ex vivo in six healthy volunteers before and after phenobarbital intake (100 mg/day) for 10 days and in six healthy control volunteers. No differences were found between groups for the difference in percentage of leukotriene B4 metabolized between day 11 and day 1. The apparent enzymatic parameters, Km and Vm, of procainamide oxidation did not differ significantly between the groups both on day 1 and day 11. These results do not show any evidence of inducibility of leukotriene B4 and procainamide metabolism in human polymorphonuclears. However, a positive correlation between 6 beta OH cortisol excretion and percentage of leukotriene B4 metabolized was observed on day 11. This study suggests that human polymorphonuclears cytochrome P450 leukotriene B4 omega-hydroxylase and procainamide metabolism is not a useful method to study cytochrome P450 induction in clinical pharmacology. PMID- 8406296 TI - Chlordiazepoxide potentiates R-phenylisopropyladenosine-induced inhibition of insulin secretion. PMID- 8406297 TI - Prediction from plasma pools in pharmacokinetic studies: a comment. PMID- 8406298 TI - High-risk groups and screening strategies for early detection of hepatocellular carcinoma in patients with chronic liver disease. AB - Characteristics of high-risk groups for hepatocellular carcinoma (HCC) in Japan were studied to establish screening strategies for early detection of the tumor. Some 93% of patients with HCC were associated with chronic liver disease. On the other hand, 67% of patients with liver cirrhosis had HCC at autopsy. Most were related to current hepatitis virus infection. An analysis of risk factors among 120 patients with chronic hepatitis revealed that age and histological findings were independent risk factors, while HBsAg, anti-HCV, sex, history of heavy drinking, history of blood transfusion were not independent risk factors. Multivariate analysis of 239 patients with liver cirrhosis demonstrated that age, positivity for HBsAg and/or anti-HCV, family history of liver disease, hepatic reserve, and a history of radical resection were independent factors related to the development of HCC. A screening schedule for cirrhotic patients was established in accordance with these results; ultrasonography was done every three months, and tumor markers measured every two months. The screening strategy proved to be effective for the early detection of HCC and improvement of the prognosis. PMID- 8406299 TI - Chemoembolotherapy for recurrent hepatocellular carcinoma in the residual liver after hepatectomy. AB - The efficacy of transcatheter arterial chemoembolization using Lipiodol (TACE) to treat recurrent hepatocellular carcinoma (r-HCC) in the residual liver after radical hepatic resection was evaluated. During the last 8 years, TACE was performed in 68 patients with r-HCC for an aggregate total of 150 times. Of the 68 patients, 4 had a massive type r-HCC with tumor thrombus in the main portal vein (PVTT) at the time of the first TACE. Among the remaining 64 patients without PVTT, multiple r-HCCs were revealed in 46, and a single r-HCC in 18 by angiography and/or follow-up CT scans after the initial TACE. In 26 of the 68 patients (38.2%), at least one or more r-HCCs were fed not only by the hepatic arteries, but also by the extrahepatic collateral arteries, such as branches of the right inferior phrenic artery. The cumulative survival rates of these patients after hepatectomy and after the initial TACE for r-HCC were 98.6% and 87.1% for one year, 89.7% and 62.9% for 2 years, 74.0% and 34.3% for 3 years, 53.1% and 20.0% for 4 years and 40.3% and 0% for 5 years (mean survival duration: 1,647 days and 947 days), respectively. These results indicate that repeat TACE against r-HCC can help obtain long-term survival in patients with r-HCC. However, during TACE, we must give consideration to the newly developed collateral feeding artery to the r-HCC. PMID- 8406300 TI - Percutaneous ethanol injection therapy for recurrent hepatocellular carcinoma. AB - Recently, small hepatocellular carcinoma (HCC) has been detected by following up patients with chronic liver diseases using ultrasonography (US). Most of the HCC patients have liver cirrhosis, and multinodular lesions are often found, even in cases of small HCC. Thus, surgical resection for small HCC is of limited efficacy. Our follow-up study indicated that a recurrent lesion was observed in 96.1% of small HCC cases after successful percutaneous ethanol injection therapy (PEIT). It is therefore very important to follow-up patients initially treated with PEIT with US for the detection of newly developed tumor nodules, because of the multicentric nature of HCC. A recurrence is subsequently again treated with PEIT. The 5-year survival rate of our 250 small HCC cases treated by PEIT is 46.7%. However, cases not followed by regular US after the first PEIT show significantly poorer 5-year survival rates. In future, it would appear to be of importance to search for means to inhibit or prevent recurrence. PMID- 8406301 TI - Long-term results after resection of hepatocellular carcinoma: experience of 480 cases. AB - The long-term outcome in 480 patients with primary hepatocellular carcinoma (HCC) who underwent hepatic resection between 1980 and 1990 was investigated. Overall 5 and 10-year survival rates were 44.1% and 17.8%, respectively, with a hospital mortality rate of 3.1%. The survival of patients who underwent curative resection was better than that of patients treated with noncurative resection. Tumor invasiveness, defined by the presence of vascular invasion and/or intrahepatic metastases, was a major prognostic factor for early recurrence in the patients treated with curative resection. The effect of tumor size and number on prognosis was attributable to a strong correlation with tumor invasiveness. One-third of patients with multiple lesions were considered to have multicentric disease, and their prognosis was better than that of patients with invasive lesions. The width of the resection margin did not affect the prognosis. An unfavorable effect of associated liver disease, especially cirrhosis, was prominent in the later period. A beneficial effect of anatomically systematic resection was apparent in non-cirrhotic patients with non-invasive HCC. PMID- 8406302 TI - Surgical treatment of recurrent hepatocellular carcinoma. AB - During the last 16 years, radical resection was performed in 392 patients with hepatocellular carcinoma (HCC). Eight patients died within 30 days after resection. The other 384 patients were discharged from hospital and closely followed for 6 to 195 months. By December 1991, 185 patients had developed a recurrent tumor, the 1-, 3-, and 5-year recurrence rates being 15.0%, 45.4%, and 55.3%, respectively. Ninety of the 185 patients underwent reoperation, including second hepatic resection (65 cases), cryosurgery (8 cases), resection of lung metastasis (6 cases), hepatic artery ligation and infusion chemotherapy (2 cases), intratumor ethanol injection (3 cases), microwave coagulation plus intratumor ethanol injection (2 cases), and exploration (4 cases). The survival rate of these 90 patients was significantly better than that of 95 patients who were treated by other palliative methods, the 5-year survival rate being 63.4% as compared with 28.6% after the first resection (P < 0.01), and 40.8% vs. 2.2% after recurrence (P < 0.01), respectively. These results suggest that reoperation for recurrent HCC might be an important approach to prolonging survival further after hepatic resection. PMID- 8406303 TI - Repeat hepatic resection for recurrent hepatocellular carcinoma. AB - During the last 12 years, hepatic resection was carried out on 268 patients with a diagnosis of hepatocellular carcinoma. Of these, 78% developed tumor recurrence in the remaining liver within 5 years. The indications for repeat hepatectomy were evaluated in the same way as for the first operation. Eighteen second hepatic resections and six third hepatic resections were performed 4 to 63 months after the first hepatectomy, with no operative deaths. The cumulative survival rate of these 24 patients was 87.1% at 3 years, 87.1% at 5 years and 72.6% at 7 years after the first hepatic resection, respectively. The present results show that repeat hepatectomy can be safely indicated for recurrent tumors in patients whose liver functional status has been relatively stable since the first hepatic resection. PMID- 8406304 TI - The significance of post-hepatectomy changes in polymorphonuclear elastase and endotoxin levels. AB - We studied the association between polymorphonuclear elastase and endotoxin levels and complications following hepatectomy. The blood concentrations of polymorphonuclear elastase, endotoxin, fibrinogen and C-reactive protein were examined with the aim of clarifying their involvement in postoperative complications in twenty-five patients who underwent hepatectomy. Polymorphonuclear elastase increased significantly (p < 0.01) on the second postoperative day compared with preoperative levels, and decreased on the seventh postoperative day. The difference in the polymorphonuclear elastase level with and without liver cirrhosis was significant (p < 0.05) on the second postoperative day. Endotoxin changed in a manner similar to polymorphonuclear elastase, but no positive correlation was found between endotoxin and polymorphonuclear elastase. Neither parameter showed any significant positive correlation with the volume of hepatic resection. We were unable to find any relationship between the degree of elevation of endotoxin and complication after hepatectomy; further study will be needed. PMID- 8406305 TI - Duodenum-preserving pancreatoduodenostomy. A new technique for complete excision of the head of the pancreas with preservation of biliary and alimentary integrity. AB - A new technique for complete excision of the head of the pancreas with preservation of the biliary and digestive tract is described. A series of 11 patients with chronic pancreatitis, pancreatic divisum, or a low-grade malignancy (mucus-producing pancreatic cancer) who underwent the procedure is reported. Previously described techniques for "duodenum-preserving pancreatic resection" differ from ours in two ways. First, other techniques accomplished only a "partial resection", retaining a rim of pancreas along the duodenum, and second, the pancreatoenteric anastomosis utilized a Roux-en-Y jejunal loop. Our procedure permits complete resection of the pancreatic head, which in turn permits a pancreatoduodenal anastomosis. This preserves the normal physiology of pancreatic digestive function. To assure an adequate blood supply to the duodenum, retroperitoneal vessels must be preserved by avoiding Kocher's maneuver, and the posterior superior pancreatoduodenal artery must also be preserved. This operation was successfully performed in 8 consecutive patients when these provisos were adhered to. PMID- 8406306 TI - Analysis of hepatic encephalopathy after distal splenorenal shunt--PTP image and pancreatic hormone kinetics. AB - The Warren-Zeppa selective distal spleno-renal shunt is aimed at decompressing esophageal varices while at the same time ligating left gastric, gastroepiploic and umbilical veins. But in the long term follow-up loss of shunt selectivity was observed in a number of cases (7/76), with hepatic encephalopathy resulting in 9.3%. In this study, we carried out percutaneous transhepatic portography and investigated pancreatic hormone kinetics in portal blood after arginine load, mainly in cases of hepatic encephalopathy after distal splenorenal shunt compared with non-encephalopathy cases of LC + Varix. These data suggested that as a cause of encephalopathy after distal splenorenal shunt, (1) a marked decrease in portal blood flow due to portal systemic shunt via the pancreatic vein was considered from PTP image, (2) a marked decrease in I/G ratio was considered from pancreatic hormone kinetics in portal blood. PMID- 8406307 TI - Clinical and histological aspects of chronic HCV infection and cirrhosis. AB - We studied 608 consecutive cases of anti-HCV-positive chronic liver disease. In 358 patients the diagnosis was established by needle liver biopsy. In 250 patients with liver cirrhosis the diagnosis was made on the basis of the unequivocal clinical signs and the results of imaging procedures. Chronic HCV infection is usually observed in adults or elderly patients; the age of the patients steadily increases with the progression of the illness to the more severe stages. Jaundice was infrequent in patients with chronic hepatitis or early cirrhosis; clinical symptoms and laboratory tests are of little value in differentiating CPH from CAH or in detecting early cirrhosis. Serum aminotransferases were usually only slightly elevated in all stages of the disease. Despite the mildness of the hepatic cytolysis, the progressive reduction in serum cholinesterase and albumin concentrations and the progressive increase in the serum alkaline phosphatase activity indicate progressive failure in the hepatic function in the course of the illness. The histological study showed that steatosis, follicular portal inflammation and eosinophilic changes in the hepatocytes were prominent features of chronic HCV infection. In contrast, severe piecemeal necrosis without bridging was rarely observed. PMID- 8406308 TI - Bile duct pressure, hormonal influence and recurrent bile duct stones. AB - An obstacle to the outflow from the bile duct not only increases bile duct pressure but also facilitates the formation of primary bile duct stones. The bile duct pressure, an indicator of the balance between bile inflow and outflow, was studied postoperatively under similar conditions in 123 patients, who had been operated on for bile duct stones. Secondary bile duct stones had been present in 86 patients (group CC) and primary stones in 26 (group C), while 11 without sphincter of Oddi function were used as a control group. The basal bile duct pressure was similar in groups CC and C, but significantly lower in the controls. Intraductal injection of saline solution caused a similar increase in bile duct pressure in groups CC and C, but not in the controls. Intravenous administration of secretin and somatostatin increased the bile duct pressure in groups CC and C, while a "normal" response to cholecystokinin, a decrease in bile duct pressure, was observed only in group CC. The "abnormal" response to cholecystokinin found in group C indicates a motor dysfunction of the sphincter of Oddi, which may have been responsible for, or at least contributed to, the formation of recurrent bile duct stones in this group. PMID- 8406309 TI - Tumor DNA content in gallbladder carcinoma. AB - An abnormal DNA content has been associated with an unfavorable prognosis in a variety of cancers. In this study, tumor DNA content was measured in patients with gallbladder carcinoma in order to determine whether DNA ploidy pattern was a prognostic indicator. Thirty-six patients who had had a gallbladder carcinoma resected with curative intent were analyzed. Aneuploid tumor (20 cases, 56 per cent) was significantly associated with poorly differentiated adenocarcinoma (p < 0.05), invasion beyond the muscularis propria (p < 0.01), and a high mitotic index (p < 0.0001). A significant advantage in terms of five-year survival was demonstrated in patients with diploid tumors as compared with those with aneuploid tumors (80 per cent versus 24 per cent, respectively, p < 0.005). Aneuploid tumors invading the subserosal layer had a significantly poorer prognosis than diploid tumors with similar depth of invasion (p < 0.05). However, when tumor invasion had extended beyond the serosa, no significant advantage in survival was found between patients with aneuploid and those with diploid tumors. It is concluded that DNA ploidy pattern is a valuable addition to a staging protocol for gallbladder carcinoma. PMID- 8406310 TI - Cholecystokinin does not delay gastric emptying of regular meals in healthy humans. AB - Exogenous CCK can delay gastric emptying of meals in the experimental animal and in humans. Studies using exogenous CCK, however, do not prove that the effects of exogenous peptide are of physiological importance. For this reason, the present study employed the specific CCK-antagonist loxiglumide (Rotta) to investigate the question as to whether CCK plays a physiological role in the regulation of gastric emptying. (1) Sonographic studies: Fourteen healthy subjects received a mixed solid-liquid 1,000 kcal meal made up of regular German food. Gastric emptying was calculated from antral volumes measured at 10-minute intervals. All subjects were studied twice on separate days, either with or without i.v. infusion of 1.0, 5.0 or 10.0 mg/kg h loxiglumide, each different dose being given to 4 or 5 subjects. (2) Scintigraphic studies: Eight subjects received an 800 kcal mixed solid-liquid meal consisting of regular food. Solid and liquid components were labeled with 99m-Tc and 113m-In, respectively. Gastric emptying was monitored using a computerized gamma-camera. All subjects were studied twice on separate days either with or without i.v. infusion of 5 mg/kg h loxiglumide. Both meals significantly increased plasma activity of CCK by 4-8 pM/ml (bioassay). However, loxiglumide did not significantly alter gastric emptying at any postprandial time when compared with the NaCl control. The failure of the CCK antagonist to affect gastric emptying was seen with both type of meals and both the sonographic and scintigraphic techniques. In the scintigraphic studies, gastric emptying of both liquid and solid components were virtually identical in experiments with or without loxiglumide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406311 TI - Recurrence of exocrine pancreatic cancer--local or hepatic? AB - The results of radical surgery for pancreatic cancer depend, like all other cancer surgery, on the technique used, and the biology of the disease. We have analysed the site and time of recurrence after pancreatectomy for exocrine pancreatic cancer in 74 patients who died more than six months postoperatively. All patients had recurrent disease, 64 patients had local recurrence in the pancreatic bed and 68 had liver metastases. Local recurrence without liver metastases was found in six patients, and ten had liver metastases, but no local recurrence. Both the time from operation to clinically evident recurrence and the postoperative survival time were significantly longer for patients with local recurrence only. Although not statistically significant, there was a tendency (5 out of 8) for smaller, well-differentiated tumors without spread outside the pancreas to be associated with local recurrences without liver metastases. We conclude that, in retrospect, the surgical procedures used were inappropriate and inadequate. To cure these patients, a more radical operation and/or effective adjuvant treatment is needed. PMID- 8406312 TI - Biliary extracorporeal shockwave lithotripsy: short-term and long-term observations in an animal model. AB - The short- and long-term effects of biliary extracorporeal shockwave lithotripsy (ESWL) using an electromagnetic lithotriptor were investigated in 26 pigs. After implantation of single human gallstones into their gallbladders, all but 4 control pigs were subjected to 4,000 or 8,000 shock waves and killed one day (n = 9), one week (n = 7), or one year (n = 6) thereafter. Post-ESWL, no abnormalities of chest radiographs or laboratory tests were detected. Apart from focal injury of the gallbladder and liver, in 4 out of 9 pigs subpleural pulmonary hemorrhages were found one day post-ESWL. However, tissue damage was largely reversed within one week and after one year only small hepatic scars persisted as permanent damage. Stone fragmentation occurred in 19 (86%) out of 22 pigs, and was adequate (fragments < or = 5 mm) in 9 (41%) pigs. Tissue damage and stone fragmentation after 4,000 as compared with 8,000 shock waves were not significantly different. These data warrant further evaluation of this lithotriptor in human studies. PMID- 8406313 TI - Endoscopic sphincterotomy in acute gallstone pancreatitis and cholangitis: a Saudi hospital experience. AB - Thirty-five patients with acute gallstone pancreatitis and/or cholangitis underwent endoscopic sphincterotomy. In 15 out of 18 patients with acute gallstone pancreatitis, common bile duct (CBD) stones, 5 of which were impacted, were removed. This resulted in prompt improvement in 12 of these patients. Eleven patients had acute gallstone cholangitis in ten of whom--including one patient who had cholangiocarcinoma associated with Clonorchis sinensis--CBD stones were extracted. Six other patients had both acute pancreatitis and cholangitis, one of whom had a choledochal cyst and had surgery; another patient with a post-surgical CBD fistula and retained stone, improved following stone extraction and nasobiliary tube insertion. The duration of hospitalization ranged between 5 and 19 days in the 26 patients treated endoscopically, and between 25 and 90 days in the 9 surgically treated patients. PMID- 8406314 TI - Spontaneous liver hematomas induced by anti-coagulation therapy. A case report and review of the literature. AB - We report on a case of massive bleeding into the liver parenchyma during treatment with a combination of warfarin sodium and trimethoprim sulfamethoxazole. A fifty-five-year-old woman was put on long-term anticoagulant therapy with warfarin sodium. Two years later a course of trimethoprim sulfamethoxazole was given to treat bronchitis. Following a bout of severe epigastric pain, ultrasonography and computed tomography (CT) then showed an enlarged liver containing several large hematomas. Subsequent CT scans, after tentative treatment only, showed regression of the liver hematomas, with almost complete disappearance after eight months. Bleeding complications and drug interactions related to this case are discussed, together with a review of the only six previous reports in the world literature of liver hematomas following anticoagulant therapy. Also mentioned are five patients in whom thrombolytic therapy gave rise to the same adverse reaction. PMID- 8406315 TI - Right anterior hepatic artery arising from the superior mesenteric artery: a case report. AB - An unusual case of right anterior hepatic artery arising from the superior mesenteric artery is presented in a patient with hepatocellular carcinoma. This variation was demonstrated by angiography and computed tomography after transcatheter arterial embolization (TAE). It has not previously been identified in the medical literature. PMID- 8406316 TI - Growth hormone and bone. AB - The significance of growth hormone in the psychology of bone remodeling is now strongly recognized. It influences osteoblast function and growth in vitro as well as in vivo. Although in vivo bone resorption is also stimulated, short administration of growth hormone induces a sustained stimulation of osteoblastic function, whereas the influence on resorption probably diminishes earlier. Growth hormone concentrations in plasma decline with age. Growth hormone and growth hormone response to growth hormone releasing factor are influenced by sex hormones, thereby changing around the menopause. In several aspects features of aging resemble those of a state of growth hormone deficiency. This holds true for body composition, the immune system as well as bone density. Therefore, it has been argued that declining growth hormone function, along with other factors, might be a causal factor in osteoporosis. Consequently, growth hormone substitution in the elderly might be applicable to maintain and possibly improve bone density and structure. The first clinical trials using recombinant growth hormone give hopeful results. PMID- 8406317 TI - Induction of islet cell surface and insulin antibodies in Balb/c mice by application of porcine insulin and Freund's adjuvant is not associated with insulitis. AB - To investigate whether insulin antibody (IAB) formation is associated with the appearance of islet cell cytoplasmic antibodies (ICA), islet cell surface antibodies (ICSA) and insulitis Balb/c Bln mice were immunized with porcine insulin in combination with or without Freund's adjuvant. The animals received 8 i.p. injections and were followed up to 150 days for the development of antibodies and insulitis. Mice immunized with insulin in CFA developed IAB as well as ICSA. Mice only receiving Freund's adjuvant emulsified in saline also developed ICSA. ICA were not detectable. Inflammatory infiltrates were found in the exocrine pancreatic parenchyma but not in islets. The results show that nonspecific stimulation of the immune system and the application of insulin as antigen leads to both the formation of ICSA and IAB, while insulitis was not detectable. PMID- 8406318 TI - Effects of chronic glipizide treatment on the NIDD heart. AB - Extrapancreatic activity of the sulfonylurea, glipizide, was evaluated in the neonatal streptozotocin-induced rat model of noninsulin-dependent diabetes. Two day old Wistar rats were given a bolus of streptozotocin (90 mg/kg i.p.) to cause noninsulin-dependent diabetes; these animals became severely glucose intolerant and eventually developed a cardiomyopathy characterized by reduced heart rate, contractility and cardiac output. Male littermates injected with citrate buffer served as nondiabetic controls. At four weeks of age, the nondiabetic and NIDD rats were administered by gavage either glipizide (2.5 mg/kg) or the methyl cellulose vehicle. Throughout the treatment protocol, no difference in the degree of glucose intolerance was observed between the glipizide-treated and vehicle treated animals. Glipizide therapy also was ineffective in improving plasma insulin levels, which were significantly depressed in the diabetic group. Yet, animals treated with glipizide for one year exhibited improved myocardial contractile function relative to the vehicle-fed or ad lib fed diabetic animals. Heart rate was significantly elevated and there was a tendency for both the rate of relaxation and contractility to be elevated in sulfonylurea-treated group. Glipizide also reduced the degree of insulin resistance in the heart. Since these changes occur in the absence of changes in glucose tolerance or insulin levels, the heart appears to be very sensitive to the direct effects of the sulfonylureas. PMID- 8406319 TI - Anabolic steroid (nandrolone) treatment during adolescence decreases the number of glucocorticoid and estrogen receptors in adult female rats. AB - The density of thymic glucocorticoid receptors and the density of uterus estrogen receptors (I) decreased significantly in adult female Wistar rats following nandrolone treatment at the age of six and seven weeks. There was no significant alteration of thymic glucocorticoid receptor density in males. Affinity of glucocorticoid and estrogen receptors did not differ from the controls significantly. The results demonstrate that the receptors of immature cells can be imprinted in adolescence similarly to those in the perinatal period and chemical substances capable of binding to the immature receptors can result in prolonged alteration of their binding character. This phenomenon should be taken into consideration especially in the case of nandrolone administration which can be used as a doping substance. PMID- 8406320 TI - Assessment of bioactivity of WHO/NIH research standard for inhibin (code 86/690) using two separate pituitary cell-culture systems. AB - The Research Standard for inhibin, porcine (No. 86/690) distributed by the National Institute for Biological Standards and Control, U.K. for the bioassay of inhibin was tested for its bioactivity in vitro in two rat pituitary cell culture systems. The system A was obtained from pituitaries of 12 day old immature rats while system B was obtained from pituitaries of mature male rats. The inhibin preparation failed to inhibit basal secretion of FSH in the system A. Instead it stimulated the release of both LH and FSH. 1 ng LHRH induced release of LH was inhibited by 32, 80 and 43% by 0.1, 1 and 10 IU inhibin, respectively. 10 ng LHRH induced release of FSH was inhibited in a none-dose related manner and the maximum inhibition was by 10 IU inhibin (25%). The same dose of 10 IU inhibin stimulated LHRH-induced release of LH by 35%. In system B, 1, 10, 50, 100 and 200 IU inhibin suppressed basal secretion of FSH by 0, 44, 72, 70 and 48%, respectively, while LH was suppressed by 13, 24, 46, 22 and 21% respectively. The pattern of inhibition of 10 ng LHRH induced release of LH and FSH by inhibin was similar to its effect on basal secretion. A specific dose-related inhibition of FSH was not observed. Our data question the utility of the inhibin standard (86/690). PMID- 8406321 TI - Insulin antibodies: reduction in various anti-insulin IgG subclasses with human insulin therapy. AB - The insulin-induced antibodies decrease on substitution of bovine-porcine with human insulin. In order to test whether there is a preferential reduction of certain anti-insulin IgG subclass, we investigated the composition of insulin antibodies. In serum samples collected from 23 bovine-porcine insulin-treated patients with clinical manifestations of insulin-induced immune problems (12 insulin resistant cases, 11 with lipodystrophy and/or local insulin allergy), the amounts of 125I-bovine insulin bound with IgG1, IgG2, IgG3 and IgG4 subclasses were 27.0 +/- 4.7, 7.8 +/- 2.3, 39.0 +/- 11.6 and 55.1 +/- 10.9 ng/ml respectively. Thus, there was an abundance of insulin antibodies of IgG4 subclass, and a relative paucity of IgG2. When the bovine-porcine insulin was changed to rDNA human insulin in 15 patients, the total insulin-binding IgG antibodies decreased by 58.6% with concomitant reductions in IgG1 (from 26.7 +/- 6.6 to 16 +/- 5.3), IgG3 (from 5.9 +/- 3.1 to 4.3 +/- 2.1), and IgG4 (from 46.4 +/- 15.6 to 18.4 +/- 7.9 ng of insulin bound/ml) subclasses. These observations suggest that in a selected subset of patients with insulin-induced immune problems, the bovine-porcine insulin induces antibodies of all four IgG subclasses but the IgG4 response predominates. With the substitution to human insulin, a reduction in the immune stimulus occurs followed with decreases in the IgG1, IgG3 and IgG4 subclasses. PMID- 8406322 TI - Omental and epigastric adipose tissue lipolytic activity in human obesity. Effect of abdominal fat distribution and relationship with hyperinsulinemia. AB - It has been shown that visceral obesity is associated with an increased incidence of hyperinsulinemia. In such a condition, hyperinsulinemia could be due to an increased lipolytic activity of omental adipose tissue (AT), through an enhanced portal flux of FFA. The purpose of our study was to evaluate the lipolytic activity of omental and epigastric AT obtained from morbid obese patients either with prevalently visceral or subcutaneous abdominal fat accumulation, evaluated by computerized tomography. The relationship between plasma insulin values and in vitro lipolytic activity in both tissues was studied. Thirteen visceral (VO) and 13 subcutaneous (SO) obese patients, matched for sex and body mass index, undergoing vertical banded gastroplasty, were studied. Before surgery, in each patient an OGTT was performed. During surgery, samples of epigastric subcutaneous and omental AT were obtained for evaluation of fat cell weight (FCW) and basal, noradrenaline 10(-5)M and isoprenaline 10(-5) M induced lipolytic activities. No significant differences in basal lipolysis were found between the two types of obesity, both in omental and in epigastric AT. In omental AT, a higher noradrenaline and isoprenaline induced lipolysis was observed in VO than in SO. Isoprenaline induced lipolysis of omental AT (expressed per cell surface area) correlated directly with FCW. VO patients showed plasma insulin values after OGTT significantly higher than SO patients. In the whole group of patients, independently from fat distribution, significant correlations were found between the incremental areas of the plasma insulin curve during OGTT and the noradrenaline an isoprenaline induced lipolytic activities both in omental and epigastric adipose tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406323 TI - Bezafibrate affects lipid, lipo- and apolipoprotein pattern in non-insulin dependent diabetic patients. AB - To assess bezafibrate efficacy in a diabetic population a single-blind randomized study was performed in 32 diet-resistant type IIb hyperlipidaemic non-insulin dependent (NID) diabetic patients in good metabolic control (HbA1c < 8%) compared to a placebo group. In our diabetic patients one month treatment of 400 mg/day bezafibrate lowered plasma C (-14%) and TG (-37%) and globally reduced the VLDL particles and VLDL lipids (-37% for C, -56% for TG and -25% for PL), raising VLDL C/TG ratio (+46%), redistributing TG from VLDL to LDL (+10%) and mainly in HDL (+49%), lowered LDL-C and Apo B levels and increased HDL-C together with Apo A1 (+19% and +13%) and Apo A1/Apo B (+72%). PL were raised by bezafibrate treatment and were redistributed from VLDL (-25%) to LDL (+25%) and HDL (+18%), while PL/C ratio increased in VLDL and in LDL (+18% and +50% respectively). Bezafibrate use was safe and improved the lipid pattern and the apolipoprotein and lipid distribution in the lipoproteins, producing a less atherogenic pattern in our NID diabetics. PMID- 8406324 TI - Continuous positive airway pressure treatment. Effects on growth hormone, insulin and glucose profiles in obstructive sleep apnea patients. AB - The principal nocturnal GH peak normally coincides with the first episode of slow wave sleep (SWS). Obstructive sleep apnea (OSA) patients have low nocturnal GH levels which may be explained by their poor quality fragmented sleep but other factors are possibly involved. Obesity is frequently associated with OSA, and obese patients also manifest reduced GH secretion. The mechanisms reducing GH levels in obese subjects are not understood, but hyperinsulinaemia is a suggested factor. In this study nocturnal plasma and secretory GH profiles of OSA patients were examined in relation to the quality and quantity of sleep, together with plasma glucose and insulin levels. Eight OSA patients, (BMI 32.7 +/- 2.3 kg/m2), underwent 2 night studies. For one night no treatment was given and for the other continuous positive airway pressure (CPAP) treatment was administered for the first time. Blood was collected continuously throughout each night and plasma GH, insulin and glucose profiles established in 10 min interval samples. From the plasma data a deconvolution model was used to calculate GH secretion rates. Sleep was recorded during the studies. For the non-treatment night GH levels were low and increased significantly with treatment, p = 0.008 for plasma levels and p = 0.02 for secretion rates. Treatment significantly decreased the cumulative apnea duration and increased the quantity of SWS and Rapid Eye Movement (REM) sleep (p = 0.008), but the mean insulin and glucose profiles did not differ between the two nights. Individual GH plasma and secretion rates, on treatment, showed a tendency to correlate with the amount of SWS (p = 0.09).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406325 TI - Correlation between glucose disposal and amino acids levels in hyperthyroidism. AB - Insulin is known to decrease plasma levels of both glucose and amino acids. We have designed a study to examine whether in hyperthyroidism, where insulin sensitivity is changed, correlation between glucose disposal and amino acids levels is maintained. We studied 5 normal (N) and 5 hyperthyroid (HTD) subjects, and measured glucose disposal rate and serum levels of plasma amino acids before and during insulin infusion utilizing the euglycemic hyperinsulinemic clamp technique. Insulin was effective in decreasing the levels of all amino acids in the plasma of all subjects. The mean amino acid decrease in response to insulin infusion rate of 10 mu/kg/min was 35 +/- 3% in N and 50 +/- 3% in HTD. The half maximally effective dose of insulin on glucose disposal was 43 +/- 3 and 69 +/- 9 uU/ml in N and HTD respectively (p < 0.05). The half maximally effective dose of insulin on decreasing plasma amino acids was 37 +/- 7 and 75 +/- 5 uU/ml in N and HTD respectively (p < 0.01). The concentration of insulin giving a half maximal effect on serum amino acids correlated with the half-maximally effective dose on glucose disposal rate (r = 0.72, p < 0.01). The maximal effect on total amino acid levels correlated with maximal glucose disposal (r = 0.76, p < 0.001). Similar results were observed with branched chain amino acids. We conclude that in hyperthyroidism glucose disposal and decrease in amino acids levels are closely correlated as in healthy subjects. This also confirms the association between glucose disposal and amino acids levels in a variety of insulin sensitive and resistant states. PMID- 8406326 TI - Effect of short-term hyperthyroxinemia on vitamin D metabolism in congenital hypothyroidism. AB - The circulating concentrations of vitamin D metabolites were measured in nine children (four to ten years of age) with congenital hypothyroidism on L-thyroxine therapy, before and after a short term increase (33%) in dosage. The concentrations of 25-hydroxyvitamin D and 24,25-dihydroxyvitamin D were not altered, but the concentration of 1,25 dihydroxyvitamin D was significantly higher in the serum of the children after three weeks of hyperthyroxinemia. This was associated with an increase in urinary calcium excretion. The increases in serum concentration of 1,25 dihydroxyvitamin D cannot be explained by differences in serum levels of calcium, phosphorus or parathyroid hormone. These findings differ from data obtained in adults. PMID- 8406327 TI - Diminished calcitonin secretion after ovariectomy without apparent reduction in calcitonin content in the rat. PMID- 8406328 TI - Nigericin inhibits adrenocorticotropic hormone- and dibutyryl-cAMP-stimulated steroidogenesis of cultured mouse adrenocortical tumor (Y1) cells. PMID- 8406329 TI - "Paradoxical" GH suppression by secretagogues in acromegaly? PMID- 8406330 TI - Increased level of basic fibroblast growth factor (bFGF) in sera of patients with malignant tumors. PMID- 8406331 TI - Serum 25-hydroxyvitamin D concentrations in colonic neoplasia. PMID- 8406332 TI - Influence of coronary artery bypass graft operation on plasma atrial natriuretic peptide concentrations. PMID- 8406333 TI - Maternal-fetal T4 transfer does not suffice to prevent the effects of in utero hypothyroidism. AB - It has been suggested recently that in congenitally hypothyroid infants with organification defect there is a maternal-fetal transfer of thyroxine (T4). The present study was conducted to evaluate how effective the maternal-fetal transfer is and whether the maternal T4 can prevent intrauterine hypothyroidism. The clinical, laboratory and radiological data on 271 full-term infants with persistent primary congenital hypothyroidism, detected by the national screening program, were used to assess the degree of in utero hypothyroidism. For 6 out of 50 athyroid infants, two pretreatment blood samples spotted on filter paper were available for calculating the T4 disappearance rate. Most infants with agenesis of the thyroid had very low T4 and very high levels of thyroid-stimulating hormone compared to infants with ectopic thyroid. In the athyroid infants the initial T4 declined to low and undetectable levels. Bone maturation was significantly delayed while the clinical symptomatology was more prominent in the athyroid congenital hypothyroid infants, as compared with the ectopic thyroid infants. In conclusion, there is some maternal-fetal transfer of T4. However, this transfer is insufficient to suppress the fetal levels of thyroid-stimulating hormone and prevent intrauterine hypothyroidism. PMID- 8406334 TI - Usefulness of urinary growth hormone (GH) measurement for evaluating endogenous GH secretion in acromegaly. AB - We investigated the relationship between urinary growth hormone (u-GH) and spontaneous 24-hour plasma GH secretion in 15 acromegalic patients. To measure u GH, we have developed a method based on concentrating the sample by centrifugal ultrafiltration and then performing an immunoradiometric assay using commercially available reagents. u-GH correlated well with the integrated concentration of plasma GH (r = 0.66, p < 0.02). Additionally, u-GH excretion in acromegalic patients was significantly higher than in the control group (190 +/- 100 vs. 3.89 +/- 0.56 pg/min, mean +/- SEM, p < 0.001). Immunoreactive u-GH showed the same elution pattern in Sephadex G-75 as standard or labeled hGH, proving that the substance measured in urine is authentic GH. In conclusion, u-GH appears to be a simple, noninvasive and inexpensive test for evaluating GH secretion in active acromegaly. PMID- 8406335 TI - Urinary GH secretion correlates with plasma GH levels during sleep and GHRH stimulation tests but not during the L-dopa stimulation test in prepubertal children. AB - In order to assess the value of urinary growth hormone (GH) as a reflection of central GH release, 42 prepubertal children with short stature and without organic disease were studied. A nocturnal GH profile and L-dopa and GH-releasing hormone (GHRH) tests were performed. Urinary GH was measured by means of a direct immunoradiometric method we have developed, using two monoclonal antibodies. Nocturnal urinary GH values correlated positively with plasma GH values expressed as the area under the curve (r = 0.76; p = 0.0001) or mean peak amplitude (r = 0.73; p = 0.0001). Also, urinary GH values correlated positively with peak plasma GH levels during the GHRH test (r = 0.64; p = 0.001). In contrast, no correlation was observed between peak plasma GH and urinary GH during the L-dopa test (r = 0.29; p = 0.11). This suggests a specific but as yet undetermined effect of L dopa on urinary GH secretion. PMID- 8406336 TI - Gonadotropin releasing hormone analogue and growth hormone therapy in precocious and premature puberty following cranial irradiation for acute lymphoblastic leukaemia. AB - Ten girls with early puberty secondary to cranial irradiation as a part of the treatment for acute lymphoblastic leukaemia (ALL) were treated with either gonadotropin-releasing hormone analogue (GnRHa) and human growth hormone (GH) (8 girls) or with GnRHa alone (2 girls). After 4 years of treatment, height SDS for bone age was improved in the group who received combined treatment (from -0.97 to +0.07, p < 0.001), in contrast to the 2 patients who received GnRHa alone in whom height standard deviation scores for bone age decreased (from -1.10 to -1.33). Sitting height in all patients was relatively shorter than leg length, and there was no significant alteration during the 4 years of treatment. PMID- 8406337 TI - Glucose metabolism in Ullrich Turner syndrome: long-term effects of therapy with human growth hormone. German Lilly UTS Study Group. AB - The effects of GH therapy on glucose metabolism in 72 Turner patients treated with human GH (HGH) 2, 3 or 4 IU/m2/day for 2 years are reported. OGTTs were performed at 0, 3, 12 and 24 months. The overall frequency of glucose intolerance was 9.7% before therapy and did not change under HGH. No change in HbA1c and fasting glucose values occurred. Integrated blood sugar values in the OGTT (area under the curre) did not change with 2 and 3 IU but were significantly elevated over control after 2 years with 4 IU. Insulin secretion was not significantly affected over time with 2 IU, whereas 3 and 4 IU produced significant increases which persisted after 2 years. Results indicate that glucose homeostasis is maintained under GH therapy at the expense of a compensatory increase in insulin secretion which persists at higher GH dosages. PMID- 8406338 TI - Changes in growth hormone-binding protein in girls with central precocious puberty treated with a depot preparation of luteinizing hormone-releasing hormone analogue. AB - Growth hormone-binding protein (GHBP) was studied in 11 girls with true precocious puberty, aged 7.3 +/- 0.2 years (mean +/- SE), before and after the first 6 months of treatment with luteinizing hormone-releasing hormone analogue D Trp6-LHRH. The 125I-human GH was incubated with 150 microliters of serum, bound and free GH were separated by gel filtration. The levels of GHBP increased significantly from 24.2 +/- 1.3 to 28.1 +/- 1.9% (p < 0.002, paired t test), more than expected for the normal age-dependent increase. The efficiency of LHRH-A therapy was confirmed by a decrease in growth rate and normalization of clinical and biological parameters. Our data agree with the hypothesis that the pubertal spurt is mediated by a sex-steroid-induced rise in GH concentration, and they suggest that the levels of GHBP may be related to the GH secretion and its variation with treatment. PMID- 8406339 TI - Idiopathic hypothalamus-pituitary dysfunction: review of five cases. AB - We describe 5 children, 4 girls, aged 4-14 years with evolving hypothalamic pituitary dysfunction. They had presenting features, isolated or combined, of polyuria and polydipsia (n = 3), weight gain and hyperphagia (n = 3), and growth failure (n = 1). During periods of 1-5 years per child, the following abnormalities developed: diabetes insipidus (n = 5), osmoreceptor dysfunction (hypernatraemia with absent thirst) (n = 3), hyperprolactinaemia (n = 3), growth hormone (GH) deficiency (n = 4, of whom 3 had normal linear growth), ACTH deficiency (n = 2), TSH deficiency (n = 2) and precocious puberty (n = 1, female). In 2 patients, high-resolution CT scans and MRI showed structural lesions of the hypothalamus 1.5 and 3.5 years after presentation. These were inaccessible and not biopsied. Scans in the remainder were normal. In conclusion, weight gain, impaired thirst, and hyperprolactinaemia were early features of evolving hypothalamic-pituitary dysfunction, and occurred with diabetes insipidus, accompanied by progressive anterior pituitary deficiencies. Pituitary hormone replacement with clinical and neuroradiological surveillance is important in any child with symptoms suggestive of an evolving hypothalamic lesion. PMID- 8406340 TI - Growth hormone (GH) response to combined pyridostigmine and GH-releasing hormone administration in patients with Prader-Labhard-Willi syndrome. AB - We evaluated the GH response to combined administration of pyridostigmine (PD), a cholinergic agonist, and GH-releasing hormone (GHRH) (60 mg PD given orally 60 min before the GHRH bolus) as well as baseline IGF-I concentrations in 10 patients (5 males and 5 females, age 6.0-24 years) with Prader-Labhard-Willi (PLW) syndrome, 8 prepubertal obese children (4 males and 4 females, age 5.6-12.0 years) and 9 prepubertal short normal children (7 males and 2 females, age 8.0 12.8 years). Mean GH responses to PD+GHRH were significantly lower (p < 0.0001) in the PLW patients (13.8 +/- 3.3 micrograms/l) than in the short normal children (52.2 +/- 9.0 micrograms/l) and similar to those of the obese children (14.3 +/- 3.2 micrograms/l). Mean serum IGF-I levels were significantly lower (p < 0.05) in the PLW patients (117.5 +/- 26.4 micrograms/l) than in the obese (329.3 +/- 88.0 micrograms/l) and the short normal children (214.3 +/- 38.3 micrograms/l). Two of the PLW patients had absent GH responses to PD+GHRH associated with subnormal IGF I concentrations, indicating pituitary GH deficiency. When these 2 cases were excluded from the statistical calculation, mean peak GH responses to PD+GHRH remained significantly lower (p < 0.0001) in the PLW patients (17.1 +/- 3.0 micrograms/l), while their mean serum IGF-I concentrations (143.4 +/- 71.5 micrograms/l) were not significantly different from those of the other two groups. These results indicate that patients with the PLW syndrome have a reduced or absent GH secretory reserve associated in some cases with low levels of IGF I.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406341 TI - Calcium metabolism after hemithyroidectomy. AB - It is not known whether thyroid surgery evokes changes in calcium metabolism. We therefore studied 6 patients operated upon with hemithyroidectomy for benign thyroid diseases, preoperatively and at 3 months and 1 year postoperatively. We measured changes in serum levels of intact parathyroid hormone (PTH), vitamin D metabolites, ionized calcium, phosphate, osteocalcin, thyroid hormones and bone density. Further, the dynamic function of the parathyroid glands was investigated by an oral calcium load test pre- and at 3 months postoperatively. At follow-up, all patients were euthyroid. During the oral calcium load, serum levels of intact PTH were reduced by the same degree before and after hemithyroidectomy, showing normal parathyroid function. Serum levels of 1,25-dihydroxyvitamin D declined from 86 +/- 16 to 57 +/- 4 pmol/l (p < 0.01) at 1 year after hemithyroidectomy, and serum levels of phosphate increased slightly from 0.96 +/- 0.08 to 1.06 +/- 0.08 mmol/l (p < 0.05), whereas serum levels of ionized calcium, intact PTH, osteocalcin and bone density did not change. Our results demonstrate that thyroid operation evokes changes in vitamin D metabolism in spite of normal parathyroid function. PMID- 8406342 TI - Subclinical hypothyroidism: a determinant of polycystic ovary syndrome. AB - The present study was an endeavor to explore whether and how hypothyroidism plays a role in the etiology of polycystic ovarian syndrome (PCOS). A composite picture of the hormone profile was assessed in different groups of subjects (control women and hypothyroid women with or without PCOS). Comparative analysis of the results suggests that hypothyroidism is invariably followed by a lowering of sex hormone binding globulin and an increment in the free testosterone level, but further metabolism of testosterone (T) may or may not be directed towards an overproduction of estriol (E3). The factors that dictate the route of T metabolism, and the way by which E3 acts to rescue the ovaries from the development of PCOS under the hypothyroid state are discussed. PMID- 8406343 TI - DNA replication in pancreatic islets and adrenals in diabetic Chinese hamsters. AB - The diabetic syndrome in Chinese hamsters is progressive and there is a decrease in the size of the pancreatic islets. To what extent this is due to impaired DNA replication in their islet cells is unknown. Therefore 3H-thymidine incorporation into islet cells was studied in genetically diabetic (subline L) male hamsters aged 17-20 days, 6 weeks, 3, 6 and 12 months. Because adrenal weight was decreased at almost all ages, studies of adrenal morphology and function were also performed. Age- and sex-matched normal (subline M) hamsters served as controls. Autoradiography showed progressively falling labeling with increasing age in both normal and diabetic animals in both islets and adrenals. At the earliest age studied, the diabetic subline animals were normoglycemic but showed hyperinsulinemia. DNA replication was reduced both in the islet cells and the adrenal cortex. From 6 weeks of age and onwards, the diabetic hamsters were hyperglycemic and usually hyperinsulinemic but DNA replication in both islets and adrenals was not different from that in normals. Histomorphometry showed reduced volume in both the adrenal medulla and cortex in 3-month-old diabetic hamsters. In 12-month-old male and female diabetic hamsters plasma cortisol and aldosterone and 24-hour urinary epinephrine were normal, but urinary norepinephrine and dopamine levels were excessive. Thus, in contrast to what is found in the obese hyperglycemic mouse, there was no increased islet cell DNA replication in response to the increased functional demand in the diabetic Chinese hamster.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406344 TI - Effects of immunoneutralization of endogenous opioid peptides on the hypothalamic pituitary-thyroid axis in rats. AB - Effects of opioid peptide antisera treatment on the secretion of thyrotropin (TSH) and thyrotropin-releasing hormone (TRH) in rats were studied. Anti-beta endorphin antiserum, anti-methionine-enkephalin antiserum, or antidynorphin antiserum was injected intraperitoneally and the rats were serially decapitated. TRH levels in the hypothalamus along with plasma TRH, TSH and thyroid hormone levels were measured by individual radioimmunoassay. TRH contents in the hypothalamus decreased significantly after opioid peptide antisera treatment, while its plasma levels tended to decrease, but not significantly. Plasma TSH levels increased significantly after opioid peptide antisera injection. Plasma TRH and TSH level responses to cold as well as plasma TSH level response to TRH were enhanced with treatment of antisera to these peptides. Plasma 3,3',5 triiodothyronine levels increased significantly after treatment of antisera to these peptides. From these findings it is concluded that the treatment of opioid peptide antisera stimulates TRH and TSH secretion in rats. PMID- 8406345 TI - Growth and metabolic data following growth hormone treatment of children with intrauterine growth retardation. AB - Twenty-five short children (6F, 19M) with intrauterine growth retardation were treated with daily subcutaneous injections of biosynthetic human growth hormone for 4 years. The treatment was commenced at a chronological age of 6.3 years (range 2.1-9.8). Eighteen of the patients had major dysmorphic signs of Russell Silver syndrome. During the first year they were randomised into two groups treated with either 15 or 30 IU of growth hormone/m2/week. The higher dose was administered to all the children after the first year of the trial. After the initial increase in growth velocity SDS (greater in those treated with the higher dose regimen), there was a progressive decrease although with values significantly higher than pretreatment levels (p < 0.02) in both groups. However there was no improvement in height SDS for bone age after 4 years. The triceps and skinfold thickness showed a decrease during the first years followed by a gradual increase. The body mass index improved during the 4 years although showing no difference between the two treatment groups. There was no alteration in thyroid function or metabolic indices (glucose or lipid homeostasis) during the study. PMID- 8406346 TI - Toward diagnostic criteria for autoimmune hepatitis. PMID- 8406347 TI - What should we clinicians do for healthy, asymptomatic, HBsAg-positive carriers? PMID- 8406348 TI - Five-year follow-up of a prospective randomized trial of hepatitis B recombinant DNA yeast vaccine vs. plasma-derived vaccine in children: immunogenicity and anamnestic responses. AB - In a prospective randomized trial, 318 children aged between 3 mo and 11 yr who were negative for all hepatitis B markers were randomized to receive two 5 micrograms doses of hepatitis B recombinant DNA yeast vaccine at 0 and 1 mo (group 1), three 5-micrograms doses of hepatitis B recombinant DNA yeast vaccine at 0, 1 and 6 mo (group 2) or three 10-micrograms doses of plasma-derived hepatitis B vaccine (group 3). The HBs antibody response rate at 8 mo was between 93% and 99%; it was still 75% to 87% at 5 yr in all three groups. Geometric mean titers at 1 yr were 83, 1,085 and 858 mIU/ml in groups 1, 2 and 3, respectively. These values had decreased after 5 yr to 47, 131 and 250 mIU/ml. Subjects in group 1 showed a significantly less proportional drop in geometric mean titer at the fifth year than did subjects in group 2 (p = 0.05) or group 3 (p = 0.015). None of the children developed HBc antibody, even after 5 yr of follow-up. We noted 42 episodes of significantly increased HBs antibody titers, probably due to anamnestic response, even when the titers had dropped to low levels. The mean age at which anamnestic response occurred was 8.7 yr.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406349 TI - High carrier rate after hepatitis B virus infection in the elderly. AB - An outbreak of hepatitis B virus infection occurred in a nursing facility; it involved 31 patients with sequelae of cerebral vascular accidents (15 men and 16 women; mean age, 77.4 +/- 9.3 yr). HBsAg disappeared within 6 mo in 9 patients and persisted during an observation period of more than 6 mo in 13; the remaining 9 patients were lost to follow-up while they carried HBsAg. Thus 13 of 22 patients followed (59%) became HBsAg carriers. We amplified a part of the S gene (436 nucleotides) with polymerase chain reaction on hepatitis B virus DNA from 12 randomly selected patients. The sequences of nine patients were the same as that of a nursing assistant who was an HBsAg carrier and suspected as the source of infection; it differed by only 1 or 2 (< 0.5%) nucleotides from those of the remaining three patients. Between the group of nine patients with transient HBV infection and the 13 patients with persistent HBV infection, we found no differences in age or sex or in parameters of nutrition or immunocompetence. These results indicate a high incidence of HBV carrier state in the elderly. PMID- 8406350 TI - Pilot study of recombinant human granulocyte-macrophage colony-stimulating factor in the treatment of chronic hepatitis B. AB - Recombinant human granulocyte-macrophage colony-stimulating factor is being used to improve the immunological function of patients with various diseases and to ameliorate hematological disorders. We investigated the tolerance and possible antiviral effect of the administration of daily doses of recombinant human granulocyte-macrophage colony-stimulating factor (3, 1 or 0.5 micrograms/kg body wt) to nine patients with chronic hepatitis B, alone or in combination with 5 MU interferon-alpha 2b. Recombinant human GM-CSF reduced significantly (p < 0.02) hepatitis B virus DNA levels. The three doses used were equally effective. Of the eight patients who completed the study, four became negative for HBV DNA and HBeAg; two of them seroconverted to HBe antibody. These four patients showed improvement in the histological activity of their liver disease. Ultimately, two patients regained normal ALT values. 2',5'-Oligoadenylate synthetase activity increased significantly (p < 0.01) in cell lysates of mononuclear cells cultured in vitro, coinciding with the reductions in hepatitis B virus DNA levels. Recombinant human granulocyte-macrophage colony-stimulating factor was well tolerated but produced a dose-dependent increase in white blood cell counts. It became intolerable at doses of 3 micrograms (and was reduced to 1.5 microgram); this effect was reversible after cessation of recombinant human granulocyte macrophage colony-stimulating factor treatment. No remarkable variations occurred in other parameters. In conclusion, recombinant human granulocyte-macrophage colony-stimulating factor administration is safe and tolerable at doses of 0.5 to 1 microgram/kg body wt and may exert an antiviral effect in chronic hepatitis B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406351 TI - Hepatitis B virus replication in diverse cell types during chronic hepatitis B virus infection. AB - Hepatitis B virus-specific nucleic acid sequences and proteins have been detected in extrahepatic tissues of acutely and chronically infected patients. However, apart from peripheral blood mononuclear cells and bone marrow cells, little is known about the specific cell types that permit viral replication. In this study, we assessed the extrahepatic tissues of four patients who died with chronic hepatitis B virus infection and two uninfected controls by means of in situ hybridization and immunohistochemical study. Three of these patients had diffuse extrahepatic distribution of the virus. Hepatitis B virus nucleic acid sequences and proteins were detected in the lymph nodes, spleen, bone marrow, kidney, skin, colon, stomach, testes and periadrenal ganglia. The following cell types were found to be positive for hepatitis B virus: endothelial cells, macrophages/monocytes, hematopoietic precursors, basal keratinocytes, mucosal epithelial cells, stromal fibroblasts and sustentacular and neuronal cells. It is probable that these cells could support viral replication because hepatitis B virus DNA replicative intermediates, viral transcripts and HBsAg and HBcAg proteins were detected in most. These findings may be relevant to the initiation of extrahepatic syndromes associated with chronic hepatitis B virus infection such as vasculitis, glomerulonephropathy, neuropathy and dermatitis. PMID- 8406352 TI - Mortality follow-up of the 1942 epidemic of hepatitis B in the U.S. Army. AB - The hypothesis that adult infection with the hepatitis B virus in the United States leads to a carrier state with a high risk of primary liver cancer was tested in two ways: (a) a cohort mortality study of U.S. Army veterans given yellow fever vaccine contaminated with hepatitis B virus in 1942 and controls and (b) a case-control study comparing veterans with hepatocellular carcinoma in Veterans Affairs hospitals with matched controls with respect to receipt of contaminated vaccine in 1942. Three groups totaling 69,988 men were the subjects of the cohort study: group 1 comprised men hospitalized with hepatitis in 1942, group 2 comprised men subclinically infected in 1942 and group 3 comprised controls who entered service after the contaminated vaccine was discontinued. Hepatocellular carcinoma cases (n = 24) and control subjects (n = 63) derived from Veterans Affairs hospital discharge files were the subjects of the case control study. Group comparisons of death rates from liver cancer were refined by expert review of records to select hepatocellular carcinoma from among all causes of death so diagnosed in the cohort study. Slightly excess mortality was found for hepatocellular carcinoma in group 2 (subclinical hepatitis B) but not for group 1 (overt hepatitis B) compared with group 3 (controls) (p = 0.08). Mortality from nonalcoholic chronic liver disease was less in group 2 than in group 3. In the case-control study, the relative risk for hepatocellular carcinoma conferred by receipt of contaminated vaccine was estimated as 3.3 (p = 0.06). We conclude from the cohort study that immunocompetent adult males rarely become carriers after hepatitis B virus infection, probably far less often than the frequently assumed rate of 5% to 10%. The small excess liver cancer mortality seen in the cohort study and the results of the case-control study are consistent, nevertheless, with the now well-established etiological role of hepatitis B virus infection in liver cancer. PMID- 8406353 TI - Extrahepatic peribiliary glands express alpha-amylase isozymes, trypsin and pancreatic lipase: an immunohistochemical analysis. AB - We examined expression of alpha-amylase isozymes (pancreatic and salivary), trypsin and pancreatic lipase on the epithelium of extrahepatic peribiliary glands immunohistochemically using 53 autopsied normal extrahepatic bile ducts. Three parts of the extrahepatic bile duct (common bile duct, intrapancreatic bile duct and bile duct at the ampulla of Vater) were examined in each case. Histologically, the extrahepatic bile duct harbored branched tubular glands (extrahepatic peribiliary glands). Extrahepatic peribiliary glands were few in the common bile duct and intrapancreatic bile duct and numerous in the bile duct at the ampulla of Vater. Immunohistochemically, pancreatic alpha-amylase was expressed in the epithelium of extrahepatic peribiliary glands in 42 cases (79%). Salivary alpha-amylase was expressed in the epithelium of the glands in 38 cases (72%). Trypsin was expressed in the epithelium of the glands in 32 cases (60%). Pancreatic lipase was expressed in the epithelium of the glands in 45 cases (85%). The immunoreactivity of these enzymes was granular and located in the supranuclear cytoplasm (corresponding to the Golgi apparatus) of the epithelium of the glands. We confirmed the specificity of the immunoreactivity of these enzymes with various methods. These results suggest that extrahepatic peribiliary glands produce alpha-amylase isozymes, trypsin and pancreatic lipase and secrete these enzymes into lumens of the extrahepatic bile duct. The secreted enzymes may play an important role in the physiology of the extrahepatic bile duct and bile. PMID- 8406354 TI - Genetic predispositions for the immunological features of chronic active hepatitis. AB - To assess the frequency and genetic predispositions of concurrent immunological diseases and immunoserological markers in autoimmune hepatitis and chronic viral hepatitis, we assessed 185 patients prospectively, including 122 patients with autoimmune hepatitis and 63 patients with viral disease. Human leukocyte antigens were determined in all patients. Sixty patients (32%) had concurrent immunological diseases, and the majority of the diseases (68%) had known human leukocyte antigen associations. Although patients with autoimmune hepatitis had concurrent immunological diseases more commonly than those with viral disease (38% vs. 22%; p = 0.04), the nature of the diseases was similar in both groups, as were the frequencies of human leukocyte antigen-DR4 (42% vs. 39%; p = 0.7). The presence of human leukocyte antigen-DR4 was associated with the concurrence of immunological diseases in both autoimmune (62% vs. 33%; p = 0.01) and viral hepatitis (75% vs. 29%; p = 0.009). In autoimmune hepatitis, human leukocyte antigen-DR4 was also associated with the expression of smooth muscle antibodies and high-titer antinuclear antibodies. We conclude that concurrent immunological diseases and immunoserological markers are common in autoimmune and chronic viral hepatitis. Both conditions have a common genetic predisposition for concurrent immunological disease associated with human leukocyte antigen-DR4. The expression of smooth muscle antibodies and high-titer antinuclear antibodies is associated with human leukocyte antigen-DR4 in autoimmune hepatitis only, suggesting that this response is associated with triggering antigens and immune recognition systems that are different from those in viral disease. PMID- 8406355 TI - Clearance by the liver in cirrhosis. II. Characterization of propranolol uptake with the multiple-indicator dilution technique. AB - We studied the steady-state hepatic extraction and single-pass hepatic uptake of propranolol in isolated perfused livers from normal rats and compared these values with those of rats with carbon tetrachloride-induced cirrhosis, rats treated with chlorpromazine (an inhibitor of propranolol metabolism) and rats with acute liver injury. The kinetics of propranolol transport in the liver were characterized by means of the multiple-indicator dilution technique, and estimates of cellular influx, efflux and sequestration rate constants were obtained with a computer fit to the model of Goresky. The outflow pattern of propranolol in the hepatic veins was then resolved into throughput material, which had swept past the hepatocytes along with albumin, and returning material, which had entered the cells but returned in the outflow after escaping metabolic sequestration. The steady-state extraction of propranolol was significantly decreased in the three experimental groups compared with that in controls, but the outflow profile differed within each group. In cirrhotic animals, influx was markedly decreased and the sequestration rate constant remained unchanged; most of the propranolol in the outflow consisted of throughput material. In rats treated with chlorpromazine, the sequestration rate constant was decreased, and propranolol in the outflow was mainly returning material. In rats with acute liver injury, both influx and sequestration rate constants were decreased. Indicator dilution curves for nonsequestered tracers showed a decreased transit time for red blood cells and abnormal diffusion of albumin and sucrose into the space of Disse in cirrhotic rats compared with the other groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406356 TI - The activity of the metabolic form of hepatic phosphatidate phosphohydrolase correlates with the severity of alcoholic fatty liver in human beings. AB - Increased esterification of fatty acids to triglyceride is common to most of the mechanisms proposed to explain the causation of alcoholic fatty liver. However, it is unclear whether this is caused by increased substrate supply or whether direct stimulation of the enzymes of the esterification pathway occurs after excessive alcohol intake. The rate-limiting step in triglyceride synthesis is catalyzed by the enzyme phosphatidate phosphohydrolase, which is present in the cytosol and microsomes and is sensitive to inhibition by N-ethylmaleimide. This enzyme is physically distinct from a second form of phosphatidate phosphohydrolase that is located predominantly in the plasma membrane, is insensitive to N-ethylmaleimide inhibition and has a putative role in cell signaling. We have investigated whether the activity of the N-ethylmaleimide sensitive ("metabolic") form of phosphatidate phosphohydrolase is increased in patients with alcoholic liver disease and whether any increased activity correlates with the severity of steatosis. N-ethylmaleimide-sensitive and insensitive phosphatidate phosphohydrolase activities were measured in needle liver biopsy specimens from 42 alcoholic patients and 6 patients with primary biliary cirrhosis and in wedge biopsy specimens from 6 normal patients undergoing routine cholecystectomy. Steatosis was "scored" on coded slides from 0 to 3. N ethylmaleimide-sensitive activity was higher in alcoholic biopsy specimens scoring 3 (3.25 +/- 0.4 units/mg protein, n = 10) than in those scoring either 0 (1.21 +/- 0.2, n = 14) or 1 to 2 (1.58 +/- 0.2, n = 18), and it was also higher than in biopsy specimens from normal and primary biliary cirrhosis patients (1.65 +/- 0.3, n = 12; p < 0.0001, analysis of variance).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406357 TI - Fatal neonatal liver failure and mitochondrial cytopathy (oxidative phosphorylation deficiency): a light and electron microscopic study of the liver. AB - Mitochondrial cytopathies are multisystemic disorders of extremely variable expression due to a deficiency in oxidative phosphorylation. Cases have recently been reported in which fatal liver failure with neonatal onset was the major clinical and biochemical syndrome. In this series we reviewed the liver histology of 10 such patients who died in the first weeks of life (from 3 days to 6 mo). In six cases the diagnosis was confirmed by study of the mitochondrial respiratory chain in the muscle, liver or both; in the other four, appropriate tests were not available for diagnosis but symptoms were identical and all other diagnoses were ruled out. In all 10 cases we noted significant steatosis, mostly microvesicular; widespread hepatocytic, canalicular and bile duct cholestasis with bile thrombi and cholangiolar proliferation; and different degrees of hepatosiderosis and glycogen depletion. Fibrosis took varying forms: perisinusoidal, periportal with the formation of septa, even precirrhosis. In the two cases of infants who died, one at 5 and one 6 mo, micronodular cirrhosis was also present. Mitochondria, either densely or loosely packed, were abnormal-pleiomorphic with few or no cristae and a granular fluffy matrix. Dense, large granules were seen in two cases. The association of neonatal liver failure and hyperlactacidemia should lead to immediate examination of the respiratory chain. The expression of this mitochondrial cytopathy can be lethal, associated with severe liver damage due to the deficiency in oxidative phosphorylation. PMID- 8406358 TI - Multiple-dose pharmacokinetics of rufloxacin in patients with cirrhosis. AB - The multiple-dose pharmacokinetics of rufloxacin were investigated in 13 patients with biopsy-proven cirrhosis and in 5 healthy controls. Rufloxacin was administered once a day for 5 consecutive days, starting with a loading dose of 400 mg on day 1 and 200 mg on the subsequent days. Plasma and urinary drug concentrations were determined by high-performance liquid chromatography and a microbiological assay. A one-compartment model applied to the high-performance liquid chromatography data was used to calculate the pharmacokinetic parameters of rufloxacin. In the controls rufloxacin had a low plasma clearance (41 +/- 4 ml/min, mean +/- S.E.M.), a long half-life (30.1 +/- 3.9 hr), a large area under the plasma concentration vs. time curve (171 +/- 18 micrograms.hr/ml) and a low renal clearance (18 +/- 2 ml/min). No appreciable differences were observed in the pharmacokinetic parameters between patients with various degrees of liver function impairment (modified Child-Pugh score ranging from 5 to 13). In these patients plasma clearance was slightly reduced (-32%), but this decrease was caused by a marked reduction in renal clearance (-65%) rather than nonrenal clearance, which remained unchanged (22 ml/min in cirrhotic patients vs. 23 ml/min in controls). A significant (p < 0.01) correlation was found between creatinine clearance and both rufloxacin renal clearance (r = 0.769) and rufloxacin plasma clearance (r = 0.681). The elimination half-life and the area under the plasma concentration vs. time curve were moderately increased in cirrhotic patients (+33% and +26%, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406359 TI - Disordered hemostasis in extrahepatic portal hypertension. AB - To assess the contribution of naturally occurring portal-systemic shunts to the coagulopathy of patients with liver disease, we studied laboratory parameters of hemostasis in 20 adult patients with extrahepatic portal hypertension, secondary to portal vein thrombosis, that had resulted in variceal bleeding. All extrahepatic portal hypertension patients had normal liver function and histological appearance. None had any evidence of preexisting coagulation disorders, and none had bled or undergone sclerotherapy in the 6 mo before study. Age- and gender-matched groups of 20 healthy individuals and 20 stable patients with cirrhosis and portal hypertension who had a history of variceal bleeding served as controls. Both patient groups had thrombocytopenia consistent with hypersplenism and portal hypertension. Prothrombin international normalized ratio (extrahepatic portal hypertension, 1.3 +/- 0.12; cirrhosis, 1.7 +/- 0.2; control, 1.02 +/- 0.06; p < 0.05) and partial thromboplastin time ratios (extrahepatic portal hypertension, 1.12 +/- 0.1; cirrhosis, 1.26 +/- 0.2; controls, 1.01 +/- 0.03; p < 0.05) were significantly prolonged in both patient groups. Extrahepatic portal hypertension and cirrhotic patient groups had significantly increased levels of serum total fibrin(ogen)-related antigen (extrahepatic portal hypertension, 818 +/- 150 ng/ml; cirrhosis, 454 +/- 52 ng/ml; controls, 124 +/- 7.3 ng/ml; p < 0.05), fibrin monomer (extrahepatic portal hypertension, 168.8 +/- 16.9 ng/ml; cirrhosis, 115.6 +/- 11.1 ng/ml; controls, 19.7 +/- 0.4 ng/ml; p < 0.05) and D-dimer (extrahepatic portal hypertension, 118 +/- 9.6 ng/ml; cirrhosis, 129 +/- 10 ng/ml; controls, 53.2 +/- 1.6 ng/ml; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406360 TI - Halothane hepatitis patients have serum antibodies that react with protein disulfide isomerase. AB - Clinical and laboratory evidence suggests that the fulminant liver failure sometimes associated with the inhalation anesthetic halothane may be an immune mediated toxicity. Most importantly, the vast majority of patients with a clinical diagnosis of halothane hepatitis have serum antibodies, which react with one or more specific liver microsomal proteins that have been covalently altered by the trifluoroacetyl chloride metabolite of halothane. The serum antibodies are specific to halothane hepatitis patients and are not seen in sera of patients with other types of liver pathology. In this study, a 57-kD trifluoroacetylated liver microsomal neoantigen associated with halothane hepatitis and native 57-kD protein were purified from liver microsomes of halothane-treated and -untreated rats, respectively. When the purified trifluoroacetylated 57-kD and native 57-kD proteins were used as test antigens in an enzyme-linked immunosorbent assay, serum antibodies from halothane hepatitis patients (n = 40) reacted with both of these proteins to a significantly greater extent than did serum antibodies from control patients (n = 32). On the basis of its apparent monomeric molecular mass, isoelectric point and NH2-terminal amino acid and tryptic peptide sequences, the 57-kD protein has been identified as rat liver protein disulfide isomerase. Antibodies raised against rat liver protein disulfide isomerase also reacted with a protein of approximately 58-kD in human liver microsomes. The results of this investigation suggest that trifluoroacetylated protein disulfide isomerase is one of the immunogens associated with halothane hepatitis. In certain patients it might lead either to specific antibodies or, possibly, to specific T cells, which could be responsible for halothane hepatitis. PMID- 8406361 TI - Zonal distribution of protein-acetaldehyde adducts in the liver of rats fed alcohol for long periods. AB - Acetaldehyde, a highly reactive intermediate of alcohol metabolism, has been shown to form adducts with liver proteins in rats fed alcohol for long periods. In this report, the zonal distribution of liver protein-acetaldehyde adducts that formed in vivo was studied by means of histoimmunostaining. Rats were pair-fed alcohol-containing and alcohol-free AIN'76 liquid diets for 2 or 11 wk before they were killed and subjected to whole body perfusion with paraformaldehyde. Each liver was cut into 60-microns-thick slices. Slices were first treated with 10% hydrogen peroxide to eliminate endogenous peroxidase activity. They were then incubated sequentially with rabbit antihemocyanin-acetaldehyde adduct, goat antirabbit serum IgG and rabbit peroxidase-antiperoxidase complex. The liver slices were stained with diaminobenzidine and counterstained with methylgreen. In the livers of rats fed alcohol for 2 wk, peroxidase activity was evident in the perivenous zone but not the periportal zone. No staining was obtained when the primary antibody had been preabsorbed with immobilized hemocyanin-acetaldehyde adduct or if the liver slices were incubated with the unimmunized rabbit IgG. Slight staining of the perivenous zone was seen in the livers of control rats, presumably because of minimal protein-acetaldehyde adduct formation emanating from endogenous acetaldehyde. When rats were fed alcohol for longer periods (e.g., 11 wk), protein-acetaldehyde adducts were still seen predominantly in the perivenous zone, but the distribution pattern was more diffuse than that observed in the livers of rats fed alcohol for only 2 wk. More liver cells produced protein-acetaldehyde adducts when rats were fed the alcohol-containing diet supplemented with cyanamide.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406362 TI - Stimulation of mono-ADP ribosylation in rat liver plasma membranes after long term alcohol intake. AB - ADP ribosylation is considered one of the important covalent modifications of cellular proteins catalyzed by ADP ribosyltransferase, which transfers ADP ribose moiety of NAD to an acceptor protein. Because a growing body of evidence has suggested significant biological roles for mono-ADP ribosylations in transmembrane signal transduction and other cell metabolism, how alcohol intake alters them is of interest. Cholera toxin and pertussis toxin have been widely used as probes to investigate the roles of GTP-binding proteins (G-proteins) in the transduction of hormonal and sensory signals. We first tested effects of long term alcohol intake on these toxin-catalyzed ADP ribosylations of G-proteins in rat liver plasma membranes. Treatment of rat liver plasma membrane with [32P]NAD and thiol-preactivated cholera toxin resulted in the labeling of a 44-kD band, most likely an alpha-subunit of the stimulatory GTP-binding protein, the extent of which was much greater in alcohol-fed rats than in pair-fed controls. Analogous experiments with pertussis toxin also demonstrated enhancement of toxin catalyzed ADP ribosylation of the inhibitory GTP-binding protein after long-term alcohol intake. More interesting was that long-term alcohol intake remarkably stimulated endogenous mono-ADP ribosylation of a 58-kD protein in a GTP-dependent manner. In vitro, ethanol (50 mmol/L) or a single load of ethanol (3 gm/kg) did not stimulate the reaction. Thus long-term alcohol intake stimulated both toxin catalyzed and endogenous mono-ADP ribosylations of proteins in rat liver plasma membranes. Pursuit of alcohol interaction with mono-ADP ribosylation may provide an interesting approach to the study of alcohol's effects on the liver. PMID- 8406363 TI - Regulation of iron metabolism in HepG2 cells: a possible role for cytokines in the hepatic deposition of iron. AB - In chronic inflammation it is reported that serum iron is depleted and hepatic iron is increased because of reticuloendothelial system iron blockade. However, recent studies indicate that hepatic parenchymal cells increase the uptake of transferrin-bound iron after in vivo stimulation with bacterial lipopolysaccharide, suggesting that endotoxemia itself or lipopolysaccharide induced production of inflammation-related cytokines may also be responsible for this phenomenon. In this study the actions of inflammation-related cytokines on the synthesis of iron-binding proteins (transferrin and ferritin) and transferrin receptor and the uptake of transferrin-bound iron were investigated in a human hepatoblastoma cell line, HepG2, which is the most commonly used cell line for examining the regulation of hepatic protein synthesis by cytokines. The cells were exposed to interleukin-1 beta, interleukin-6 or tumor necrosis factor-alpha separately for 24 hr. In each cytokine treatment group, the level of transferrin, which is secreted into the conditioned medium, was found to be decreased compared with that of untreated cells. On the other hand, the biosynthesis of ferritin was markedly elevated after the same treatment. This increase in ferritin by cytokine treatment was diminished when deferoxamine was used concomitantly to deplete intracellular chelatable iron. After stimulation with interleukin-1 beta, interleukin-6 or tumor necrosis factor-alpha, 59Fe-labeled transferrin uptake into the cells was increased by 36%, 48%, or 18%, respectively, and this uptake was inhibited by the addition of excess unlabeled transferrin. A binding study with 125I-labeled diferric transferrin revealed that the three cytokines increased the number of transferrin receptors on the cell surface by 1.15-fold to 1.35-fold.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406364 TI - Oxidative stress produced by suprahepatic occlusion and reperfusion. AB - In this article the spontaneous chemiluminescence and the steady-state concentration of hydrogen peroxide were determined in rat liver as indicators of oxidative stress in the tissue. Hydroperoxide-initiated chemiluminescence and the activity of antioxidant enzymes (catalase, superoxide dismutase and glutathione peroxidase) were also measured to evaluate antioxidant defenses and serum activity of lactate dehydrogenase and aspartate aminotransferase. Mitochondrial morphology and mitochondrial respiratory control ratio were measured as indicators of cell and mitochondrial damage. Xanthine dehydrogenase and xanthine oxidase activities were determined as a possible source of oxyradicals. No significant changes were observed after 10 or 30 min of vena cava occlusion in any of the measured parameters. In contrast, 10 min of occlusion followed by 10 min of reperfusion increased chemiluminescence (from 18 +/- 3 to 32 +/- 5 cps/cm2), hydrogen peroxide (from 0.10 +/- 0.01 to 0.17 +/- 0.01 mumol/L), lactate dehydrogenase (from 80 +/- 2 to 330 +/- 30 U/L), and aspartate aminotransferase (from 42 +/- 2 to 100 +/- 10 U/L). Liver reperfusion was also associated with mitochondrial swelling and decreased mitochondrial respiratory control (from 5.6 +/- 0.3 to 2.6 +/- 0.1). The activity of the antioxidant enzymes and xanthine oxidase was instead without change. After 30 min of vena cava occlusion and 10 min of reperfusion a more marked increase in chemiluminescence (37 +/- 5 cps/cm2), hydrogen peroxide (0.30 +/- 0.01 mumol/L), lactate dehydrogenase (730 +/- 10 U/L) and aspartate aminotransferase (140 +/- 10 U/L) was observed. No further changes were found in either mitochondrial morphology or respiratory control (2.4 +/- 0.1) in isolated mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406365 TI - Metabolic adaptation of the kidney to hyperammonemia during chronic liver insufficiency in the rat. AB - The aim of this study was to evaluate the role of renal ammonia and glutamine metabolism in the metabolic adaptation to chronic liver insufficiency-induced hyperammonemia in the rat. To this purpose, urinary excretion, renal net exchange and tissue concentrations of ammonia and amino acids were measured in anesthetized, normal control rats that did not undergo surgery, in control rats that underwent sham surgery, in rats that underwent portacaval shunting and in rats that underwent both portacaval shunting and bile duct ligation. Rats that underwent sham surgery and portacaval shunting were pair-fed with rats that underwent portacaval shunting and biliary obstruction, to correct for anorexia in that group, and all rats that were operated on were studied 7 and 14 days after surgery. Arterial ammonia and glutamine levels were elevated in groups that underwent portacaval shunting and portacaval shunting plus biliary obstruction at all time points. At days 7 and 14, total renal ammonia production decreased in rats that underwent portacaval shunting and in rats that underwent portacaval shunting plus biliary obstruction, associated with a 50% decrease in net renal glutamine uptake and strongly diminished net ammonia release into the renal vein, which was most prominent in the group that underwent portacaval shunting plus biliary obstruction. Urinary ammonia excretion was similar in rats that underwent portacaval shunting and in those that underwent sham surgery but was increased more than 200% at days 7 and 14 in rats that underwent portacaval shunting plus biliary obstruction. In this group, in contrast to portacaval-shunted rats, the kidney appeared to be an organ of net ammonia disposal from the body. In separate experiments in unanesthetized, unrestrained rats, similar changes in urinary ammonia excretion were observed without changes in arterial pH, excluding an effect of anesthesia or pH on the obtained results. These results indicate that the kidney plays an important role in the metabolic adaptation to hyperammonemia during chronic liver insufficiency in the rat. PMID- 8406366 TI - Microtubule-dependent transport of bile salts through hepatocytes: cholic vs. taurocholic acid. AB - Studies with taurine-conjugated bile salts have demonstrated two pathways for hepatocellular delivery of bile salts to bile: a cytosolic, microtubule independent pathway and a membrane-based, microtubule-dependent pathway. However, a significant portion of circulating bile salts may be unconjugated. To determine whether free bile salts utilize similar pathways, we examined the effect of colchicine on the biliary excretion of intravenously administered cholic acid and taurocholate in intact rats. Basal rats were pretreated with low-dose colchicine or its inactive isomer, lumicolchicine, 1 hr before placement of intravenous and biliary cannulas and 2.75 hr before intravenous injection of [14C]cholic acid and [3H]taurocholate. Superfused rats were prepared as above but with intravenous infusion of taurocholate at 200 nmol/min.100 gm beginning 0.75 hr before [14C]cholic acid/[3H]taurocholate injection. Depleted/reinfused rats were subjected to biliary diversion for 20 hr before colchicine or lumicolchicine pretreatment, infusion of taurocholate and [14C]cholic acid/[3H]taurocholate injection. In each group, biliary excretion of [14C]taurocholate and [3H]taurocholate was inhibited equally by colchicine; for peak excretion rates the respective inhibition values were 33% and 35% in basal rats, 63% and 65% in superfused rats, and 74% and 76% in depleted/reinfused rats. Biliary excretion of [14C]taurocholate occurred consistently later than excretion of [3H]taurocholate, and maximal rates of excretion were reduced. In contrast, plasma uptake rates of [14C]cholic acid and [3H]taurocholate were essentially the same in depleted/reinfused rats. Deconvolution analysis of [14C]taurocholate vs. [3H]taurocholate biliary excretion curves revealed no significant differences among experimental groups. We conclude that conversion of [14C]cholic acid to [14C]taurocholate slightly retards its biliary excretion and diminishes its peak excretion rate compared with exogenous [3H]taurocholate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406367 TI - Relationship between genomic DNA ploidy and parameters of liver damage during necrosis and regeneration induced by thioacetamide. AB - Thioacetamide proved to be a potent necrogenic agent when a single dose of 6.6 mmol/kg was administered intraperitoneally to rats. Its necrogenic ability was assessed on the basis of morphological and biochemical changes. The injury of centrilobular hepatocytes showed a peak of cell death 24 hr after thioacetamide administration; it was followed immediately by the regenerative response. Parallel increases of serum aminotransferases, isocitrate dehydrogenase and gamma glutamyl transferase activities were observed. Severe liver damage was also evident at 24 hr on the basis of glutathione depletion (29% of control), malondialdehyde production (169%), cytochrome P-450 level decrease (26%) and increased activity of glutathione S-transferase (160%). We checked the regenerative response by determining nuclear DNA content in isolated hepatocytes 0, 6, 12, 18, 24, 36, 48 and 72 hr after thioacetamide administration. Changes in DNA cell distribution between G0-G1, S and G2 + M phases of the cell cycle were observed. The sharp decrease in the percentage of the tetraploid cell population (G2 + M phases) and the abrupt increase of the S-phase cells at 36 and 48 hr suggest transition from adult to fetal in hepatocyte populations obtained 24 and 36 hr after thioacetamide treatment. At 72 hr of treatment, hepatocyte populations showed recovery to adult state. In the shift from the adult to fetal, registered at 24, 36 and 48 hr after thioacetamide administration, mitosis seemed to precede the synthesis of DNA. PMID- 8406368 TI - Origin and differentiation of hepatic natural killer cells (pit cells). AB - Liver sinusoids contain a population of large granular lymphocytes or natural killer cells, originally termed pit cells. After isolation and purification, these cells were separated into a low-density and a high-density fraction. The liver low-density fraction differs significantly in morphology and function from cells of the blood, whereas the liver high-density fraction shows intermediate properties. In this study we demonstrate that this morphological and functional heterogeneity is based on subsequent steps of differentiation of the large granular lymphocytes within the liver. When cell proliferation was suppressed by sublethal total body irradiation, the life span of the hepatic large granular lymphocytes could be determined: high-density and low-density populations were totally depleted within 1 and 2 wk after irradiation, respectively. By using intravenous asialo-GM1 anti-serum to deplete animals of asialo-GM1-positive cells, we found that the depletion of the asialo-GM1-positive cells preceded the depletion of asialo-GM1-negative hepatic low-density large granular lymphocytes by approximately 1 wk. Direct evidence that the asialo-GM1-positive high-density large granular lymphocytes are precursors of the low-density large granular lymphocytes was given by adoptive transfer experiments with fluorescent-labeled high-density cells. Three days after their injection, labeled large granular lymphocytes were found in the hepatic low-density fraction of the recipient rat, and these cells had developed morphological characteristics of low-density large granular lymphocytes. It is concluded therefore that marginating blood large granular lymphocytes differentiate through high-density large granular lymphocytes into the typical liver specific low-density large granular lymphocytes or pit cells. PMID- 8406369 TI - Quantitative detection of hemoglobin saturation in the liver with near-infrared spectroscopy. AB - To quantify the changes in oxygen saturation of hemoglobin in the liver in hypoxia and liver transplantation, we applied a novel method using near-infrared spectroscopy. Instead of the conventional two-wave-length method, we obtained near-infrared data from a wide spectral range of 700 nm to 1,000 nm with continuous-wave spectroscopy. To correct the flattened spectral shape caused by photon scattering in living tissue, we then applied an equation taking into account the relationship between absorber concentration and actual absorption in scattering materials as assessed with time-resolved spectroscopy. Hepatic hemoglobin oxygen saturation was calculated with multicomponent analysis. In room air, hepatic hemoglobin oxygen saturation of rabbit was calculated as 57.4% +/- 2.5% (mean +/- S.E.M., n = 7). Arterial, portal and hepatic venous hemoglobin oxygen saturation were simultaneously measured as 97.5% +/- 0.7%, 77.1% +/- 3.4% and 55.5% +/- 4.6%, respectively. The changes in hepatic hemoglobin oxygen saturation seen in graded hypoxia were also close to those in hepatic venous hemoglobin oxygen saturation, suggesting that the average oxygenation state of sinusoidal blood approximates that in the central vein. We tested the clinical applicability of this method in a case of liver transplantation. It was determined that the hepatic hemoglobin oxygen saturation of the graft liver was heterogeneously distributed and that the initially low level of hepatic hemoglobin oxygen saturation was increased by the ligation of portal-systemic shunts. PMID- 8406370 TI - Contribution of sinusoidal endothelial liver cells to liver fibrosis: expression of transforming growth factor-beta 1 receptors and modulation of plasmin generating enzymes by transforming growth factor-beta 1. AB - Transforming growth factor-beta 1 is an important cytokine in the pathophysiology of liver fibrosis, stimulating the production of extracellular matrix. Whether this cytokine can also control the degradation of matrix proteins in liver cells has not been investigated. Because plasmin is an important protease for the degradation of matrix glycoproteins, we investigated whether sinusoidal endothelial liver cells could contribute to fibrosing liver disease through the modulation of plasmin-generating enzymes in response to transforming growth factor-beta 1. Sinusoidal endothelial cells from guinea pig liver were investigated in pure monolayer culture. Using 125I-labelled transforming growth factor-beta, we demonstrated high-affinity binding sites on sinusoidal endothelial cells at a density of 9.3 x 10(2) per cell, and a dissociation constant of about 5.5 x 10(-11) mol/L. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the known three classes of membrane receptors for transforming growth factor-beta. Using biosynthetic labeling of proteins with 35S methionine, immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, we showed that sinusoidal endothelial cells produce and secrete plasminogen activator inhibitor type 1 from the beginning of culture. Treatment of confluent cell cultures for 24 hr with transforming growth factor-beta 1 increased synthesis and release of plasminogen activator inhibitor type 1. The response was almost maximal at a concentration of 1 ng transforming growth factor beta/ml and paralleled the increased synthesis of fibronectin. On reverse fibrin autography we proved that transforming growth factor-beta 1 stimulated the release of functionally active plasminogen activator inhibitor type 1.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406371 TI - Centrilobular distribution of acetaldehyde and collagen in the ethanol-fed micropig. AB - We established a new animal model of alcoholic liver disease in the micropig, a species that consumes ethanol voluntarily in the diet. Ten micropigs were pair fed diets containing 40% of calories as ethanol or cornstarch with identical amounts of fat, protein and micronutrients for 12 mo. Liver histopathology in the ethanol-fed pigs included steatonecrosis in all five and interstitial and perivenous fibrosis in three. Electron microscopy showed Ito-cell transformation with perisinusoidal collagen accumulation. Acetaldehyde adducts were found by immunofluorescence in the centrilobular region and were focused in perivenous zone 3 of all ethanol-fed animals. Protein and triglyceride levels were increased, whereas vitamin A and iron levels were decreased in liver homogenates from ethanol-fed animals. Thus, in this new animal model of alcoholism, ethanol feeding produced the features of alcoholic liver disease concurrent with hepatic deficiency of selected nutrients. Histological and immunofluorescent studies provide in vivo evidence that perivenous collagen deposition is linked to ethanol metabolism and acetaldehyde production. PMID- 8406372 TI - Endothelins 1 and 3: potent cholestatic agents secreted and excreted by the liver that interact with cyclosporine. AB - Autoradiographic studies have shown that the liver accumulates endothelin. High affinity binding sites for endothelin have been identified on rat liver plasma membranes. We investigated the role of endothelin isopeptides as mediators of cholestasis with isolated rat liver perfused by a recirculating solution of buffer and blood. These studies demonstrated that endothelin-1, as measured by means of radioimmunoassay, was cleared from the perfusate by the liver and that the liver concentrated both endothelin-1 and endothelin-3 in bile. Addition of endothelin-1 to the liver perfusate solution increased the concentration of endothelin-3 measured in the perfusate, suggesting that endothelin-1 caused release or secretion of endothelin-3. Both endothelin-1 and endothelin-3 at 5 nmol/L caused almost complete cessation of bile flow, but this effect was more prolonged after endothelin-1 than after endothelin-3 administration. Because it has been reported that cyclosporine increases endothelin levels, we studied the interaction of these two compounds. Cyclosporine (100 mumol/L) also produced cholestasis. Endothelin-3 secretion in bile, however, was decreased in livers perfused with cyclosporine compared with secretion in controls. Simultaneous addition of endothelin-1 and cyclosporine that on their own were not significantly cholestatic produced cholestasis. In conclusion, endothelin is a potent cholestatic agent secreted and excreted by the liver. It may potentiate the cholestatic action of cyclosporine. PMID- 8406373 TI - Hepatic blood volume responses and compliance in cats with long-term bile duct ligation. AB - Hepatic capacitance responses were compared in sham-operated and 14-day bile duct ligated cats under pentobarbital anesthesia. Both groups were subjected to splenectomy and had the anterior hepatic nerve plexus sectioned to allow stimulation; the posterior plexus was intact. Blood volume compensation for hemorrhage was reduced in the bile duct ligation group compared with the control group: The liver compensated for 20.1% and 10.6% of blood loss, respectively. Portal hypertension did not exist and hepatic compliance was unaltered despite the presence of severe biliary hyperplasia, portal tract distortion and fibrosis. The capacitance responses to hemorrhage in the bile duct ligation group were accounted for entirely by a passive compliant response to reduced portal pressure, whereas the sham surgery group showed an additional active component. Responses of liver volume and venous resistance to norepinephrine were normal, but responses to nerve stimulation were reduced. The nerve dysfunction was not universal; reflex arterial blood pressure response to carotid occlusion was normal. The data suggest that hepatic blood volume responses to hemorrhage is mediated by passive compliant responses to reduced portal pressure (stressed volume) and active responses (unstressed volume) to nerve stimulation but that long-term bile duct ligation produces selective hepatic neuropathy resulting in loss of the active component. PMID- 8406374 TI - Des-gamma-carboxy (abnormal) prothrombin and hepatocellular carcinoma: a critical review. AB - Des-gamma-carboxyprothrombin (DCP) appears to be a useful tumor marker for the evaluation of patients with HCC. DCP is produced by the malignant hepatocyte and appears to result from an acquired posttranslational defect in the vitamin K dependent carboxylase system. DCP production is independent of vitamin K deficiency, although pharmacological doses of vitamin K can transiently suppress DCP production in some tumors. DCP levels greater than 0.1 AU/ml (100 ng/ml) on ELISA are highly suggestive of HCC or tumor recurrence. Normalization of DCP levels correlates well with successful tumor resection and appears to be an excellent marker of tumor activity. Plasma DCP does not correlate with AFP levels. However, when used together, DCP and AFP assays increase the sensitivity to HCC in more than 85% of patients. The specificity of the DCP assay appears to be superior to that of AFP; fewer than 5% of patients with nonmalignant liver disorders have DCP levels in excess of 100 ng/ml. In patients with medium to large HCC, DCP levels do correlate with tumor size. In tumors of less than 3 cm, DCP levels are increased in only 20% of patients. However, the diagnostic threshold for the DCP assay may be improved by newer assays that can detect partially carboxylated DCP species not measured by the monoclonal antibody-based ELISA. PMID- 8406375 TI - Meeting report: International Autoimmune Hepatitis Group. PMID- 8406376 TI - Is anything better than MOPP? PMID- 8406377 TI - Subcutaneous low dose arabinosyl-cytosine and oral idarubicin in high risk adult acute myelogenous leukemia. AB - In order to further explore low dose chemotherapy for high risk acute myelogenous leukemia (AML), low dose Ara-C and oral idarubicin (LAI) were given to 33 patients of 24-84 (median 66) years with AML after myelodysplastic syndrome (MDS) (12 patients), refractory AML (13 patients), and AML with contraindications to intensive chemotherapy (8 patients). Patients received 1 to 4 cycles of Ara-C 10 mg/m2 q 12 h s.c. inject. on days 1-14 and idarubicin 20 mg/m2/d orally days 3, 4, 5. Three Three patients attained complete remission, four patients partial remission and one patient minor response, whereas 11 patients succumbed to early mortality from hemorrhage (two patients) and/or infections (10 patients). Three of 13 patients with heavily pretreated refractory AML went into remission compared to 3/12 with AML after MDS and 1/8 with AML and contraindications against intensive treatment. Median duration of CR is 102 (70-488 +) days. Thirty two of 33 patients developed grade 4 hematological toxicity requiring platelet transfusions. The non-hematologic toxicity was acceptable. LAI provides a standardized therapeutic option especially for heavily pretreated patients with AML. PMID- 8406378 TI - Identification of some marker chromosomes in acute leukemias by fluorescence in situ hybridization. AB - Four patients with acute leukemia are presented in whom conventional karyotypic analysis had revealed complex numerical and structural changes including unidentifiable (marker) chromosomes. It is demonstrated that the application of fluorescence in situ hybridization (FISH) may contribute to the identification of such anomalies. PMID- 8406379 TI - Pharmacokinetics of high dose methylprednisolone and use in hematological malignancies. AB - The pharmacokinetics of oral and intravenous high dose methylprednisolone (Solu medrone, Upjohn) were compared in patients with hematological malignancies. The aim of the study was to determine the oral bioavailability of high dose methylprednisolone and to establish whether this is a feasible and more convenient route of administration. The plasma pharmacokinetics were described by a one-compartment open model with peak plasma levels of 6.9 +/- 2.5 micrograms/ml. Total area under the plasma concentration versus time curve was similar by either route. Mean relative oral bioavailability was generally high (91 +/- 27 per cent). Retrospective analysis of 34 patients with chronic lymphocytic leukemia (CLL), non-Hodgkin's and Hodgkin's lymphoma treated with high dose methylprednisolone showed 11 responses including two complete remissions among nine patients with CLL. There was significant improvement in platelet counts in thrombocytopenic patients and treatment was well tolerated and toxicity was relatively low. High dose methylprednisolone may therefore be a useful palliative treatment for hematological malignancies, particularly where marrow suppression is a problem. PMID- 8406380 TI - Evidence of colony suppressor activity and deficiency of hematopoietic growth factors in hairy cell leukemia. AB - The cause of myelosuppression in hairy cell leukemia (HCL) has been ascribed to a reduction of the circulating progenitor cell (CPC) compartment and to suppression of hematopoiesis by TNF-alpha. The present study was performed to evaluate the inhibitory effect of hairy cells (HCs) and a possible lack of hematopoietic growth factors on the number of autologous CPCs in vitro. In initial experiments the number of circulating BFU-E, CFU-GM and CFU-mix in HCL patients was found decreased. Monocytopenia but not the number of circulating HCs correlated to the degree of colony reduction in our patients. This pointed to a lack of colony stimulating factors (CSFs) in HCL. Actually, the growth of BFU-E, CFU-GM, and CFU mix improved upon the addition of IL-3 and GM-CSF in HCL patients but not in healthy donors. To test the suppressive role of HCs in our assay system, cultures were performed after removal of autologous HCs. The results showed that in HC depleted cultures the numbers of BFU-E, CFU-GM, and CFU-mix were significantly higher. This inhibitory effect of HCs could partially be neutralized by the addition of monoclonal antibodies against TNF-alpha. When the assays were performed with the removal of HCs and the addition of CSFs normal progenitor cell counts were detected in most patients. We conclude that HCs mediate the inhibition of colony growth in part by TNF-alpha. Monocytopenia is related with a deficiency of CSFs in this disease. The reduced colony growth in HCL, therefore, is due to both the inhibitory effects of HCs and the deficiency of CSFs. We suppose that the CPC-compartment is actually preserved in this disease. PMID- 8406381 TI - Primary neoplasia of the female peritoneum. AB - Primary neoplasms of the female peritoneum may be mesothelial or Mullerian in nature. Primary mesotheliomas may be cystic, benign papillary or diffuse malignant. Primary Mullerian tumours of the peritoneum are thought to arise within the secondary Mullerian system: the overwhelming majority are serous in nature and probably develop from pre-existing endosalpingiosis. Primary peritoneal serous tumours of borderline malignancy are identical histologically to the peritoneal 'implants' found in association with ovarian serous tumours of borderline malignancy, whereas primary peritoneal serous adenocarcinomas are histologically identical to ovarian serous adenocarcinomas. The recognition of a primary peritoneal serous neoplasm of borderline malignancy rests on the presence of either normal ovaries, ovaries containing only a fully benign neoplasm or ovaries showing only minimal surface involvement. The diagnosis of a primary peritoneal serous adenocarcinoma is made if the ovaries are of normal size with either no tumour or only minimal surface involvement: in some cases normal ovaries will have previously been prophylactically removed because of a family history of ovarian cancer. The differential diagnosis between a primary serous adenocarcinoma and a diffuse malignant mesothelioma can be difficult and is often not resolved by either electronmicroscopy or immunocytochemistry. PMID- 8406382 TI - Granulomatous hidradenitis suppurativa and cutaneous Crohn's disease. AB - Three patients with concurrent hidradenitis suppurativa and Crohn's disease are presented. The notable histological feature in each hidradenitis resection was the presence of numerous discrete epithelioid granulomas in areas of non-inflamed dermis. The purpose of the study was to determine the incidence of epithelioid granulomas in 101 hidradenitis patients and their significance in relation to systemic granulomatous disease. Discrete epithelioid granulomas were identified in 8% of the resections (10 patients). One patient had Crohn's disease and one other pulmonary sarcoidosis. Seven patients with granulomatous hidradenitis neither had nor developed any other disease during the 3-year follow-up period. Clinical review identified a further two patients with Crohn's disease but associated with non-granulomatous changes in the skin resections. Foreign body type granulomas were identified in 25% of the resections adjacent to ruptured hair follicles, sinus tracts or nearby degenerate sweat glands. The study shows that, although foreign body type granulomas are a common finding in hidradenitis, the presence of discrete epithelioid granulomas in the dermis away from the site of active inflammation is unusual and should alert the pathologist to the possibility of systemic granulomatous disease such as Crohn's disease or sarcoidosis. PMID- 8406383 TI - Cutaneous immunocytomas: a clinicopathologic study of 26 cases. AB - The clinical and histological features of 16 patients with a primary cutaneous immunocytoma and 10 patients with a secondary cutaneous immunocytoma are reported. In all cases the diagnosis was based on the presence of monotypic plasma cells or lymphoplasmacytoid cells. Our data show that primary cutaneous immunocytomas are a distinct type of cutaneous lymphoma, characterized by (a) the presence of solitary or localized skin lesions (13 of 16 cases); (b) preferential localization on arms and legs (15 of 16 cases); (c) excellent response to local treatment (15 of 16 cases) and (d) a favourable prognosis. Histologically, these primary cutaneous immunocytomas are characterized by the presence of nodular or diffuse infiltrates with monotypic lymphoplasmacytoid/plasma cells located at the periphery of the infiltrates. Important clinical and histological differences were noted between primary and secondary immunocytomas. In the latter group more widespread skin disease was seen, often in the presence of paraproteins and/or autoimmune diseases. In contrast with the peripheral localization of the monotypic cells in primary cutaneous immunocytomas the monotypic lymphoplasmacytoid/plasma cells in secondary immunocytomas formed diffuse infiltrates or these cells were found dispersed throughout the infiltrate. There were no differences in clinical presentation or course between the different subtypes of cutaneous immunocytomas (lymphoplasmacytic, lymphoplasmacytoid and polymorphic immunocytomas). The differential diagnosis between primary cutaneous immunocytomas and cutaneous plasmacytomas, primary follicular centre cell lymphomas and cutaneous 'pseudolymphomas' is discussed. PMID- 8406384 TI - Differences in clinical behaviour and immunophenotype between primary cutaneous and primary nodal anaplastic large cell lymphoma of T-cell or null cell phenotype. AB - The histological, immunophenotypic and clinical features of 19 primary cutaneous anaplastic large cell lymphomas (cutaneous ALCL) were compared with those of 18 primary nodal anaplastic large cell lymphomas (nodal ALCL) of T-cell or null cell type. Although cutaneous ALCL and nodal ALCL had identical morphological features, differences in surface marker expression and clinical behaviour were found. Immunophenotypical differences concerned the expression of epithelial membrane antigen (82% of the nodal ALCL were positive v. none of the cutaneous ALCL) and the cutaneous lymphocyte antigen (HECA-452), a possible skin-homing receptor on cutaneous T-lymphocytes (most tumour cells in 44% of cutaneous ALCL cases were positive, whereas nodal ALCL showed expression of HECA-452 on only few tumour cells (< 25%) in 18% of cases tested). Loss of T-cell markers was more pronounced for nodal ALCL. Patients with cutaneous ALCL were generally older (median 61 years) than patients with nodal ALCL (median 24 years) and, in contrast to the latter group, did not show bimodal age distribution. Survival after 4 years, using lymphoma-related death as an end-point, differed significantly between cutaneous ALCL and nodal ALCL; 92% for cutaneous ALCL and 65% for nodal ALCL (P = 0.04). The better survival of cutaneous ALCL patients could not be ascribed to differences in age, stage or initial mode of treatment. These data indicate that differences in immunophenotype and clinical behaviour exist between morphologically identical primary cutaneous and primary node-based ALCL. They indicate that the primary site is an important prognostic factor in predicting the clinical outcome of ALCL. PMID- 8406385 TI - The nature of atypical multinucleated stromal cells: a study of 37 cases from different sites. AB - The morphology, immunophenotype and ultrastructure of atypical multinucleated stromal cells in 37 specimens from different anatomical sites were compared. Overall, cellular morphology was similar between sites but nuclear features, immunophenotype and ultrastructure varied and reflected that of adjacent normal mononuclear stromal cells. It is concluded that the atypical multinucleated stromal cells represent a reactive change in the indigenous stromal cells. In all the cases mast cells were seen as a prominent component of the inflammatory infiltrate and many of them were intimately related to the atypical stromal cells. It is suggested that reactive stromal cell changes may relate to an interaction with mast cells. PMID- 8406386 TI - Lymphomas of mucosa-associated lymphoid tissue arising in the urinary bladder. AB - The clinical, histological and immunohistochemical findings in five primary lymphomas of the urinary bladder are reported. One patient had both lymphoma and transitional cell carcinoma of the bladder. All of the lymphomas showed histological features of mucosa-associated lymphoid tissue (MALT) lymphomas with centrocyte-like cells in all cases. One patient with pre-existing cystitis glandularis showed lymphoepithelial lesions. Biopsies from four patients contained reactive germinal centres and, in two of these, there was follicular colonization by tumour cells. In three patients, repeat biopsies, over several years, showing the changes of MALT lymphoma, were diagnosed as cystitis. We suggest that a large proportion of primary lymphomas of the bladder are lymphomas of MALT and that the characteristic morphological and immunohistochemical features of these tumours should be sought in biopsies containing large numbers of lymphoid cells. PMID- 8406387 TI - Lectin binding and expression of blood group-related antigens in carcinoma-in situ and invasive carcinoma of urinary bladder. AB - To determine whether histochemical reactivities of carcinoma-in-situ of the urinary bladder differ from those of invasive transitional cell carcinoma, we tested a profile of eight different lectins and three antibodies directed against blood group-related antigens for 15 cases of carcinoma-in-situ and 26 cases of non-papillary (6 superficially and 20 deeply) invasive transitional cell carcinoma that had been diagnosed according to the histopathological criteria of the International Union against Cancer. For biotin-labelled lectins and monoclonal antibodies to mouse blood group-related antigens, the avidin-biotin peroxidase complex method was applied. Positive histochemical reactions of peanut agglutinin without neuraminidase treatment--PNA N(-)--in the 20 deeply invasive tumour cases were significantly higher than those in the 15 carcinoma-in-situ cases (P < 0.05). In contrast, the reactions of blood group-related antigens in the 20 deeply invasive tumour cases were significantly lower than those in the 15 carcinoma-in-situ cases or the 11 normal controls (P < 0.05). The results confirm previously reported studies of the staining of PNA N(-) and blood group-related antigens on carcinoma-in-situ and invasive tumours of urothelial organs. The application of lectins and blood group-related antigens to the histopathology of urinary bladder cancer may be helpful in the differential diagnosis of carcinoma in-situ from invasive cancer, but neither PNA N(-) nor blood group-related antigens can be solely reliable in this. PMID- 8406388 TI - An immunophenotypic survey of malignant melanomas. AB - An immunohistochemical study of 106 malignant melanoma specimens from 59 patients, using formalin-fixed, paraffin-embedded material, is reported. Negativity for HMB-45 was seen in 11% of specimens. The rate of positivity with CAM 5.2 was 7%. One specimen showed alpha-smooth muscle actin (alpha SMA) positivity. For 11 of the 12 cases in which anomalous immunophenotypes were seen, multiple specimens were available; nine of these showed evidence of an alteration in the immunophenotype between specimens. Comparing the primary tumours with local recurrences and metastases, there was, variously, loss of HMB-45, S-100 protein and NKI/C3 positivity, and acquisition of CAM 5.2 and alpha SMA positivity. In some cases, the change of immunophenotype appeared to occur in a single step. However, one case with six consecutive specimens showed evidence of progressive loss of HMB-45, S-100 protein and NKI/C3 with concomitant gain of CAM 5.2 staining. The implications for the use of immunophenotyping in diagnostic practice are discussed. PMID- 8406389 TI - A comparison of the ultrastructure of pigment granules in melanosis ilei and pulmonary lymph nodes. AB - Two cases of melanosis ilei were studied, in which grossly visible blackish pigmentation of the ileal mucosa was incidentally discovered at autopsy. Light microscopy showed that the pigment granules lay within macrophages in atrophic Peyer's patches. Ultrastructural studies showed that the pigment granules were heterolysosomes containing crystalline material, particles, granules and, occasionally, lipid droplets. The morphological similarity between these pigment granules and granules in pulmonary macrophages was established through ultrastructural studies of pulmonary lymph nodes obtained during routine autopsies. These data, plus results of past electron-probe X-ray analytic studies by us and others, leads us to conclude that the pigment granules in melanosis ilei contain exogenous material derived from inspired and ingested materials. PMID- 8406390 TI - Expression of CD30 and nerve growth factor-receptor in neoplastic and reactive vascular lesions: an immunohistochemical study. AB - A total of 118 biopsies from 20 different types of benign, malignant and reactive vascular proliferations were examined for the expression of the CD30 antigen using the monoclonal antibodies BerH2 and HRS4 on paraffin-embedded sections. The results were compared with those obtained using an antibody directed against the partially homologous nerve growth factor-receptor. The vascular character of the lesions was assessed by means of endothelial markers and the expression of intermediate filaments was verified immunohistochemically. CD30 was expressed in the endothelial component of more than the half of all tumours, with the exceptions of Kaposi's sarcoma and teleangiectatic granuloma. There was a slightly higher rate of Ki-1 positivity in malignant lesions. Nerve growth factor receptor could be demonstrated in a similar percentage in both endothelium and pericytes, but no correlation with CD30 could be established. We conclude, therefore, that these antigens are not adequate for the differential diagnosis of vascular lesions, nor do they help distinguish between benign and malignant lesions. Their expression seems to reflect different states of cellular function or activation. It does not necessarily occur simultaneously or in the same cell type. PMID- 8406391 TI - KP1 (CD68) expression in benign neural tumours. Further evidence of its low specificity as a histiocytic/myeloid marker. PMID- 8406392 TI - Hepatocellular carcinoma with osteoclast-like giant cells. PMID- 8406393 TI - High incidence of Epstein-Barr virus detection in Hodgkin's disease and absence of detection in anaplastic large-cell lymphoma in children. AB - By in situ hybridization with EBER oligonucleotides and immunohistochemistry with anti-latent membrane protein 1 (LMP1) antibody, we compared the detection rate of Epstein-Barr virus (EBV) in Hodgkin's disease and anaplastic large-cell lymphomas in children. Among the 13 cases of Hodgkin's disease tested, 7 (54%) were found to be EBV associated (EBER transcripts +, LMP1 +). None of the 11 cases of ALC lymphomas was found to contain EBV genomes or gene products. This may indicate that EBV is not a pathogenic agent in anaplastic large-cell lymphomas in children in comparison to Hodgkin's disease. PMID- 8406394 TI - Tufted angioma of late onset. PMID- 8406395 TI - Estimates of nuclear volume in endometrial adenocarcinoma: the effect of good quality sections. PMID- 8406396 TI - Save the mucocoele. PMID- 8406397 TI - Demonstration of Mycobacterium tuberculosis in colonoscopic biopsies. PMID- 8406398 TI - Genograms: a psychosocial assessment tool for hospice. AB - Genograms are a valuable and non-threatening evaluation tool for hospice patients and families. The genogram provides basic information about the family including the role of each member and the family dynamics. As the diagram is drawn, family life cycle issues and relationships between family members become evident. The genogram may go beyond the household to include supportive neighbors, friends, and community resources. Religious and spiritual support is also noted. The information is used to assess family needs and to provide interventions both before the death and during bereavement. PMID- 8406399 TI - The evaluation of hospice home care volunteers. AB - A national sample of 139 hospice volunteer coordinators rated a series of statements about volunteer behaviors in terms of their importance to volunteer performance. Each statement had to be rated at least 8 on a scale of 10 by 75% of the respondents in order to be included in the final instrument. This procedure yielded a scale of 27 descriptors of appropriate volunteer behavior. The scale is intended for use by hospice volunteer coordinators who want a tool with which to offer concrete, behavior based, constructive feedback to volunteers about their performance. PMID- 8406400 TI - Achieving a healthy death: the dying person's attitudinal contributions. AB - This study addressed the question of whether there are attitudes that may be psychologically beneficial to the dying, their families, and their caregivers. The Omega Attitudes Inventory was distributed to 467 systematically selected hospice coordinators nationwide. The responses of 327 (70%) indicated high concordance with patient attitudes contributing to a "healthy" death. The identified attitudes were qualitatively enhanced through anecdote and the literature. The study concluded with implications for clinical practice and further research. PMID- 8406401 TI - Teaching Rwandan families to care for people with AIDS at home. AB - AIDS causes disabling symptoms during its chronic and terminal phases. Families throughout the world, whether related to the patient by blood or affection, provide most of the personal care for him or her at home during these phases. Whether the family has access to advanced medical care or not, they can be taught simple comfort measures and nursing care skills that will improve the well-being of the patient. In Rwanda, a small country in east-central Africa, Red Cross volunteers were trained to teach these skills. The volunteers then returned to their villages to help local families. Six months later, 24 of these families were interviewed about the impact of the volunteers' visits. Families indicated they had benefited from being taught the caregiving skills. They also appreciated the emotional support attendant to the volunteers' visits. In summary, the training course enabled volunteers both to enhance family nursing care skills and to provide emotional support to families caring for people with AIDS at home. This training can also be used by volunteers to assist families caring for members ill with other endemic chronic infectious diseases. PMID- 8406402 TI - Teaching hospice medicine to medical students, house staff, and other caregivers in the United Kingdom. AB - This paper outlines the development of a medical education programme within the existing programme of the Macmillan Education Centre of the Dorothy House Foundation, Bath, England, between 1989 and 1992. A review of some of the significant contributions to the literature in this area is followed by a description of the initial research that was instrumental in designing these courses. A review of activities of the Centre is reported. A more detailed description of a course specifically designed for family doctors is made. A natural development from that course was one for all disciplines involved in caring for the terminally ill--this is described--along with some of the difficulties of such a method of education. Reference is made to evaluative work undertaken with reporting of initial data and work still in progress. Conclusions are drawn about the nature of such workshops and comment made on the role of facilitation. PMID- 8406403 TI - Medical education for hospice care: a selected bibliography with brief annotations. AB - This is a briefly annotated bibliography of useful materials for the education of health professionals, principally physicians. It encompasses teaching goals, methods, and settings, as well as model courses, course evaluation, communication skills, general resources on death education, and miscellaneous background pieces. PMID- 8406404 TI - Comparative morphine pharmacokinetics following sublingual, intramuscular, and oral administration in patients with cancer. AB - Previous literature reports have suggested that sublingually administered morphine sulfate results in an improved bioavailability of the drug when compared to orally administered morphine. To investigate this possibility further, we studied six cancer patients all of whom received 10 mg doses of morphine sulfate by intramuscular, oral and sublingual routes. Pharmacokinetic analyses failed to suggest an advantage of sublingual administration when compared with oral dosing. Bioavailability of morphine following intramuscular administration appeared superior to both oral and sublingual routes. PMID- 8406405 TI - Chaos, population biology, and epidemiology: some research implications. AB - In this article I aim to provide some feeling of the new paradigm of disease causation (chaos) as it applies to the field of population biology and epidemiology. A secondary objective is to show, with the aid of qualitative methods, how one can approach chaos in time-series data. The multifactorial stochastic paradigm of causation is contrasted with the new deterministic approach. This approach is embedded in the theory of nonlinear system dynamics. Chaos implies that randomness is intrinsic to a nonlinear deterministic system; this is true despite the extent of knowledge of the intervening causes and, ultimately, despite determinism. Three research avenues are discussed in depth from the standpoint of chaos theory. First, the topic of sporadic epidemics is dealt with. I argue that the space-time clustering of cases from a starting epidemic is due to a sudden and high increase of the contact rate beyond a threshold. Interaction rather than main effects and nonlinear rather than linear dynamics are involved. Second, the incubation period of disease is studied. I advocate that an individual-level deterministic process underlies Sartwell's model of the incubation period. This accounts for the robustness of the model vis a-vis confounding variables. Third, monozygotic twinning is analyzed. Assumed by some to be a random process, monozygotic twinning proves to be dynamically different from dizygotic or single-maternity processes; its dynamics can actually be chaotic. Throughout the provided examples, the point is made that chancelike phenomena are primarily concerned with chaos theory. For biological problems showing recurrent inconsistencies by stochastic modeling, dynamic modeling should be envisaged. Inconsistencies can suggest that the relevant factors are out of the model and that they are related deterministically. Finally, spectral analysis and attractors in the phase space are presented; these tools can aid the population biologist in tracing out chaos from time-series data sets. Several time-series data sets are simulated according to a simple nonlinear difference equation that bears some relationship to the basics of the dynamics of infections in the population. I show how the series can be analyzed and interpreted. Much research remains to be carried out until the nonlinear effects of risk factors can be validated. The undertaking is worth the effort, as a new paradigm of causation is at stake. PMID- 8406406 TI - Multivariate fluctuating asymmetry in Israeli adults. AB - Fluctuating asymmetry serves as an indicator of developmental homeostasis; any genetic or environmental factor that destroys homeostasis disturbs bilateral symmetry. Many studies have attempted to correlate increased fluctuating asymmetry with measures of developmental homeostasis or of adaptation, but collective results have been equivocal. We still have much to learn about fluctuating asymmetry itself. We develop a multivariate treatment of bilateral asymmetry that allows us to measure and adjust for the effects of both directional (right versus left) asymmetry and antisymmetry (handedness) and for the effects of size and sex. We examine 29 morphometric traits (13 size and shape, 8 pairs of bilateral traits) on 400 Israeli adults. We show that, even if metric traits are logarithmically transformed at the outset so that nonlinear allometric relations among them are properly accounted for and if the resulting measures are adjusted for sex by subtracting the means and dividing by the standard deviations, the correlation matrices among traits are strongly heterogeneous across sexes. Although the bilateral traits as a set are modestly correlated with the size and shape traits, neither size nor shape is correlated with asymmetry. Moreover, asymmetry traits are virtually uncorrelated inter se. We also examine the often touted connection between asymmetry and departures from morphometric modality and show that asymmetry is not correlated with deviations from either average size or average shape. If properly measured, fluctuating asymmetry becomes a measure of the noise in development; in our study it correlates with nothing. We discuss these results in light of empirical results and in terms of developmental theory suggesting that fluctuating asymmetry should be correlated with measures of maladaptation. Some of the effects reported in the literature need to be reexamined with more refined morphometric and statistical techniques. PMID- 8406407 TI - Selection on maternal and neonate size at birth. AB - Neonatal size is an important factor in determining the survivorship of infants within the first month of life. Because maternal size has an influence on neonatal size, selection should operate on those characters correlated with birth weight and gestational age, including maternal prepregnancy weight, height, and age. In the present study we use a path-analysis approach to examine the operation of selection on both neonatal and maternal size. We found that neonatal survivorship depends not only on the size of the infant at birth but also on a negative allometric relationship between the size of the neonate and the size of the mother. Thus, although the size of the mother has no direct effect on neonatal mortality, the mothers of surviving infants tend to be smaller relative to the size of their neonate. This may provide a mechanism whereby selection maintains a balance between maternal size and neonate size. PMID- 8406408 TI - Age and infertility in a micronesian atoll population. AB - The relationship between female age and infertility is examined using a single island Micronesian population case. Demographic data, derived primarily from reproductive history interviews, show that a significant age-associated decline in marital reproductive performance is absent before women reach their late thirties in this population but a substantial decline is present once women reach their forties. Ethnographic data support the demographic inference that couples are maintaining relatively high levels of conjugal coital activity with both advancing female age and increasing marital duration. Thus coital activity levels appear to be an important factor in the maintenance of fertility in this group before the mid-thirties but decreases in fecundability after this age are due primarily to reductions in fecundity, not to declines in coital activity. The description of the Butaritari case lends support to Underwood's (1990) suggestion that a "Micronesian pattern" of reproductive performance may exist for the region's atoll-based populations and underscores the promise of further investigations of these special cases in the fields of demography and reproductive ecology. PMID- 8406409 TI - Use of growth models to describe patterns of length, weight, and head circumference among breast-fed and formula-fed infants: the DARLING Study. AB - Nine mathematical models were applied to 24-month length, weight, and head circumference growth curves of 39 breast-fed and 31 formula-fed infants from the DARLING study. For both breast-fed and formula-fed infants, the Karlberg infancy childhood-puberty (ICP) model best describes the relationship between recumbent length and age and the Jenss model best describes the relationship between head circumference and age. For formula-fed infants several models appropriately describe the relationship between weight and age, and we recommend the Jenss model. However, none of the models suitably describes the shape of the weight curve for breast-fed infants. PMID- 8406410 TI - Correlates of menstrual cycle length in south Indian women: a prospective study. AB - There has been a lack of agreement on the variation in and the correlates of menstrual cycle length in the literature. A total of 2566 women from rural and urban areas in one district of Tamil Nadu in southern India were studied prospectively to identify correlates of menstrual cycle length. Rural women had higher odds for short and for long cycle lengths. Younger women, those with higher age at marriage, and those with lower educational status had significantly longer cycle lengths. PMID- 8406411 TI - Extraction of DNA from frozen red blood cells. PMID- 8406412 TI - Hemopexin: a unique genetic polymorphism in populations of African ancestry. AB - Using isoelectric focusing and immunoblotting techniques, we have screened 937 plasma or serum samples from Nigerian blacks (N = 380), Papua New Guineans (N = 110), Aleuts (N = 62), Mayans (N = 139), Dogrib Indians (N = 45), and Eskimos from Kodiak and St. Lawrence islands (N = 201) for the hemopexin (HPX) polymorphism. We compared these data with our previously published data for US whites (N = 267) and US blacks (N = 194). Except for Nigerian blacks and US blacks, HPX was found to be monomorphic for the common HPX*1 allele in all populations tested. In addition to the commonly occurring HPX*1 allele, two other less common alleles, HPX*2 and HPX*3, were observed with respective frequencies of 1.8% and 4.6% in US blacks and 1.7% and 9.0% in Nigerian blacks. These data strongly suggest that the HPX*2 and HPX*3 alleles are unique alleles restricted to the black gene pool and are of potential significance in microevolutionary studies and in defining African admixture in hybrid populations. In addition to their importance in anthropogenetic studies, these unique HPX mutations also have potential biological significance in hemolytic disorders. PMID- 8406413 TI - Cytokine expression in malignant lymphomas: a complicated puzzle with many pieces to assemble. PMID- 8406414 TI - Cytokines in malignant lymphomas: review and prospective evaluation. AB - Cytokines play important roles in the pathogenesis of lymphomas. Cytokines either can be produced or exert effects on neoplastic or reactive cells. The secretion of cytokines can provide growth advantages for tumor cells in either an autocrine or a paracrine fashion. An elevated serum or tissue level of cytokines can contribute to the clinical and histopathologic alterations associated with malignant lymphomas. The effects of cytokines on the histopathologic changes are most noticeable in Hodgkin's disease (HD). The malignant (Hodgkin's-Reed Sternberg) cells in HD have been shown to secrete interleukin-1 (IL-1), IL-5, IL 6, IL-9, tumor necrosis factor-alpha, macrophage colony-stimulating factor, transforming growth factor-beta, and, less frequently, IL-4 and granulocyte colony-stimulating factor. These cytokines may be responsible for the increased cellular reaction and fibrosis observed in tissues involved by HD and for the immunosuppression in patients with HD. In contrast to Hodgkin's-Reed-Sternberg cells, most non-HD lymphoma cells do not produce cytokines in excess amounts. Exceptions include T-cell-rich B-cell lymphoma (IL-4), angioimmunoblastic lymphadenopathy-like T-cell lymphoma with plasmacytosis and hypergammaglobulinemia (IL-6), anaplastic large-cell lymphoma (IL-9), polymorphic immunocytoma (IL-6), and immunoblastic lymphoma (IBL) (IL-6). Some cytokines are involved in the unique cellular reactions in each of these types of lymphoma. For example, IL-4 is responsible for the T-cell reaction in T-cell-rich B-cell lymphoma, while IL-6 is accountable for the plasma cell reaction in angioimmunoblastic lymphadenopathy-type T-cell lymphoma. Others may be directly involved in the tumor cell growth or differentiation. For instance, IL-9 may be important for the autocrine proliferation of anaplastic large cell lymphoma, whereas IL-6 is essential for plasmacytoid differentiation in polymorphic immunocytoma. Further studies of the roles of cytokines in lymphomas may lead to major advances in the understanding of the molecular processes involved in the histopathogenesis of malignant lymphomas. Elucidation of the autocrine or paracrine function of cytokines also may lead to new approaches to a rational intervention in these disease processes. PMID- 8406415 TI - In situ localization of parathyroid hormone-like protein and mRNA in intraepithelial neoplasia and invasive carcinoma of the uterine cervix. AB - Parathyroid-like protein (PLP), or parathyroid hormone-related peptide, is a well recognized mediator of paraneoplastic hypercalcemia (humoral hypercalcemia of malignancy syndrome). In this study we examined the expression of PLP by 40 invasive squamous cell carcinomas (SCCs) of the cervix and selected carcinomas of nonsquamous histology. Using a polyclonal antibody to human PLP, 93% of SCCs, including two tumors from patients with humoral hypercalcemia of malignancy syndrome, showed moderate to strong cytoplasmic immunoperoxidase staining for PLP. The strongest staining often was observed in areas of invasion associated with stromal desmoplasia. The small number of weak or negatively stained SCCs were all poorly differentiated tumors. Although native uninvolved squamous epithelium showed weak to moderate staining of the superficial layers, there was variable or full-thickness immunostaining in areas of dysplasia. Normal endocervical glands and stroma as well as cervical adenocarcinomas and neuroendocrine carcinomas were negative. In situ hybridization studies showed abundant PLP mRNA within SCC in patients with hypercalcemia. However, PLP mRNA was of relatively low abundance in tumors of normocalcemic patients. Ultrastructural studies showed cytoplasmic, membrane-bound, granular inclusions in tumor cells from the hypercalcemic patients. Our data suggest that increased PLP gene transcription contributes to the increased production of PLP and the pathogenesis of humoral hypercalcemia of malignancy syndrome. PMID- 8406416 TI - Graft vascular disease in the great vessels and vasa vasorum. AB - To determine whether the great vessels were subject to transplantation-associated arteriosclerosis (graft vascular disease) we studied sections of aorta and pulmonary arteries from 19 cardiac explant cases. Sections of the same vessels from five autopsied hearts from patients who had received liver and/or kidney transplants with immunotherapy as well as from eight recipient hearts served as controls. In addition, eight donor hearts (three aortae and eight pulmonary arteries) that were not used for heart transplantation were examined. Intimal proliferation was noted in the aorta but affected the pulmonary arteries only slightly, whereas the vasa vasorum of both vessels were involved and were occluded or stenosed by thickened intima containing recanalized capillaries. On morphometric study the ratio of intima to media in the aorta was significantly higher in the heart transplant group than in the control group (0.14 +/- 0.14 [n = 19] v 0.02 +/- 0.02 [n = 16]; P < .01). This intimal thickening was not correlated with the use of cyclosporine or with other clinicopathologic factors. The cytomegalovirus infection rate was higher in the heart transplant group (85%) than in the control group (40%). PMID- 8406417 TI - The histopathology of uterine leiomyomas following treatment with gonadotropin releasing hormone analogues. AB - Gonadotropin-releasing hormone analogues (GnRH agonists) cause pituitary desensitization by downregulation of GnRH receptors, decrease gonadal steroid production, and reduce uterine volume in women with leiomyomas. The purpose of this study was to examine the morphologic changes in uterine leiomyomas associated with GnRH agonist treatment. The study group consisted of 33 patients (mean age, 36.9 years) who presented with infertility, dysmenorrhea, and/or menorrhagia, and who were treated with a GnRH agonist prior to surgery. A control group consisted of 44 premenopausal patients (mean age, 41.5 years) with similar symptomatology who underwent resection of leiomyomas only. In neither group was there any history of recent pregnancy, uterine surgery, or hormone replacement therapy. Microscopic review of all cases was performed without knowledge of the therapeutic history. No differences with respect to mitotic activity, fibrosis, edema, or vascular changes were detected. There is a suggestion that leiomyomas subjected to preoperative GnRH agonist treatment showed increased cellularity (P = .04); necrosis (P < .001) was associated with preoperative GnRH agonist treatment. The reduction of leiomyoma size during GnRH agonist therapy may be due to both ischemic injury and cellular atrophy. Although necrosis of leiomyomas is associated with GnRH agonist treatment, the lack of significant pleomorphism or mitotic activity distinguishes these altered leiomyomas from leiomyosarcomas. PMID- 8406418 TI - Telepathology with an integrated services digital network--a new tool for image transfer in surgical pathology: a preliminary report. AB - We describe a low-cost telepathology system working via a commercial integrated services digital network (ISDN) and consisting of modular software and hardware elements. The main elements are Apple Macintosh workstations; a software program for the simultaneous transfer of pictures, voice, and data; and procedures for image processing and general administration of all the information generated. Additionally, the system allows remote control of any peripheral instruments by a "picture-instrument manager." The transfer rate is currently 64 kbit/s; it will be extended to 128 kbit/s (ISDN basic rate) in the near future and to 2 Mbit/s (ISDN primary rate) in the next 2 years. The system was tested by the regional hospital in Samedan, Switzerland, and the Department of Pathology, University of Basel, Basel, Switzerland, a distance of 250 km, by offering a remote frozen section service to the regional hospital in 16 cases. Fifty-four to 58 seconds were needed for the transfer of a diagnostic 8-bit grey-level image containing 341 (median value) +/- 26.1 (standard error) kbytes (n = 13) or a diagnostic 24 bit color image containing 165 (median value) +/- 16.9 (standard error) kbytes (n = 3). The time required for a diagnostic session was between 25 and 35 minutes. PMID- 8406419 TI - The presence of epithelioid granulomas in hepatitis C virus-related cirrhosis. AB - We examined surgical liver specimens from 52 patients with hepatitis C virus related cirrhosis. All patients underwent orthotopic liver transplantation at Paul Brousse Hospital. They were found to be seropositive for antibodies to hepatitis C virus by second-generation testing (RIBA 2, Ortho Diagnostic Systems Inc, Westwood, MA). We detected multiple granulomas in five (10%) of the cirrhotic livers. These granulomas were composed of epithelioid cells, sometimes associated with multinucleated giant cells, and were surrounded by small lymphocytes and fibrosis. The epithelioid granulomas were located within the cirrhotic nodules. They were not present within the portal tracts or within the fibrosis. These granulomas were diffusely distributed in the liver. None of the patients with diffuse hepatic epithelioid granulomas had evidence of tuberculosis or brucellosis before transplantation or during the follow-up period (range, 3 to 20 months). They had no detectable cause of granulomatous hepatitis. The role of hepatitis C virus as a cause of epithelioid granulomas is discussed. PMID- 8406420 TI - Gastric adenocarcinoma with ciliated tumor cells. AB - Ciliated metaplasia (CM) of gastric mucosa is common in Japanese individuals but rare in whites. We studied 36 consecutive gastrectomies from Chinese individuals for the presence of CM. The gastrectomies were performed for adenocarcinoma (n = 22), benign ulcer (n = 13), and malignant lymphoma (n = 1). Ciliated metaplasia was found in 16 (44.4%) of the stomachs, with equal distribution between male and female patients. Ciliated metaplasia was present in 11 cases (50%) of adenocarcinoma and in five cases (38.5%) of benign ulcer, the difference being statistically insignificant. Contrary to previous studies, there was no increased frequency of CM in gastric mucosa harboring an intestinal-type adenocarcinoma compared with those with non-intestinal-type tumors. Ciliated metaplasia always co-existed with intestinal metaplasia. Colonization by Helicobacter pylori was not associated with an increased incidence of CM. In five cases (22.7%) of adenocarcinoma ciliated tumor cells were found, and all of these cases showed CM in the gastric mucosa. In one of these cases the whole tumor was composed of glands formed by ciliated cells. In the other cases there was a morphologically different nonciliated component. The cilia in the tumor cells showed light microscopic and ultrastructural changes identical to those of the cilia seen in CM affecting the nonneoplastic gastric mucosa. This study documents for the first time the co-existence of CM and ciliated adenocarcinoma in the same stomach. It is concluded that in the Hong Kong Chinese population CM is commonly present in gastric mucosa harboring an adenocarcinoma or benign ulcer. Adenocarcinomas with ciliated cells are strongly associated with CM. PMID- 8406421 TI - Immunohistologic studies of Kikuchi's disease. AB - An immunohistologic study of lymph nodes from 21 patients with Kikuchi's disease (histiocytic necrotizing lymphadenitis) was performed. The cell components of the affected areas were mainly CD4-positive cells, CD8-positive T cells, alpha/beta T cell gene receptor-positive T cells, and lysozyme-staining cells. CD3-positive or alpha/beta T-cell gene receptor-positive T cells were composed mainly of CD8 positive and CD11b-negative cytotoxic T cells. Double staining demonstrated that CD4-positive cells usually were positive for Ki-M1p, a marker of plasmacytoid monocytes, but negative for T-cell markers. Although some lysozyme and CD4 double positive cells were recognized, most CD4-positive cells were negative for lysozyme. The results indicate that CD4-positive cells in the affected foci of Kikuchi's disease were mainly composed of plasmacytoid monocytes. PMID- 8406422 TI - Granulosa cell tumor of the adult testis: a clinicopathologic study of seven cases and a review of the literature. AB - We report a study of seven men, aged 16 to 76 years (average age, 47.4 years) with granulosa cell tumor (GCT) of the testis. Three patients presented with testicular enlargement of several years' duration and a fourth presented with a testicular enlargement of unknown duration. The tumors in three patients were detected during routine physical examination. None of the patients had endocrine related symptoms. All tumors were well circumscribed and showed the solid, cystic, microfollicular, gyriform, insular, and trabecular patterns typical of GCT of the ovary. Call-Exner bodies were present in three tumors and two tumors had a focal spindle-cell component. In one case the surrounding testicular parenchyma showed Leydig's cell hyperplasia and a Sertoli cell nodule. The tumor cells revealed strong immunoreactivity for vimentin but showed no expression for keratin or epithelial membrane antigen. One patient developed liver and retroperitoneal lymph node metastases 121 months after initial diagnosis and died 13 months later. Another patient initially presented with retroperitoneal lymph node metastasis and developed metastasis to the inguinal lymph nodes 12 months later. Three patients are alive at 1, 4, and 37 months with no evidence of disease. Another patient died of an unrelated condition. Follow-up information was not available for the seventh patient. Twelve cases of GCT of the adult testis have been reported in the literature, with metastases occurring in two: one of these two patients had a tumor for 8 years and died of disease 5 months after diagnosis with multiple metastases and the other had metastasis at the time of diagnosis, but was free of disease for 14 years. Our findings and a review of the literature indicate that GCT of the adult testis is a rare and slow-growing neoplasm with the potential to form distant metastases. Because recurrence or distant metastasis may occur late in the clinical course, long-term follow-up of these patients is recommended. PMID- 8406423 TI - A case of fatal infectious mononucleosis presenting with fulminant hepatic failure associated with an extensive CD8-positive lymphocyte infiltration in the liver. AB - We describe a fatal case of infectious mononucleosis presenting with fulminant hepatic failure associated with extensive CD8-positive lymphocyte infiltration and diffuse karyorrhexis in the liver. Immunohistochemical analysis of mononuclear cells showed that Leu-2a (CD8)-positive lymphocytes were heavily distributed in the portal areas and the sinusoidal spaces, but Leu-3a (CD4)-, Leu 14 (CD22)-, or My 4 (CD14)-positive cells were undetectable in sections of the liver. Southern blot hybridization studies disclosed the presence of Epstein-Barr virus DNA fragments in the liver tissue. The unusual pathologic and immunologic responses observed in this case could not simply be explained by severe Epstein Barr virus infection. Some superimposed factors should be considered. PMID- 8406424 TI - Endocarditis caused by Rochalimaea henselae. AB - A case of community-acquired, culture-negative, infective endocarditis was diagnosed in a 57-year-old construction worker. Small, pleomorphic gram-negative rods were seen in Brown-Hopps tissue gram stains and Warthin-Starry silver stains. The organism was identified as Rochalimaea henselae by polymerase chain reaction amplification and sequencing of the 16S rDNA gene sequence. This is the first report of infective endocarditis caused by R henselae. PMID- 8406425 TI - Cystic dysplasia of the testis. AB - Cystic dysplasia of the testis (CDT) is a rare congenital defect that results in the formation of numerous irregular cystic spaces within the mediastinum testis. We describe a 4-year-old boy with right testicular swelling who underwent orchiectomy for CDT after a preoperative ultrasound examination revealed a multicystic, anechoic lesion. Grossly, the lesion was 2 cm in size and was composed of multiple, irregularly shaped cystic spaces lined by flattened cuboidal epithelium. Immunohistochemical studies revealed that the epithelial cells expressed keratin, vimentin, and epithelial membrane antigen. Both the histologic appearance of the cyst lining and the immunohistochemical profile resembled the epithelium of rete testis. In our review of the literature 10 cases (including this lesion) of CDT have been described. A defect in the connection between the efferent ductules, derived from the mesonephric epithelium, and the rete testis and seminiferous tubules, derived from the gonadal blastema, is currently thought to be the most likely explanation of the pathogenesis of CDT. PMID- 8406426 TI - Disseminated adenoviral infection presenting as acute pancreatitis. AB - Adenoviruses are gradually being recognized as a significant source of morbidity and mortality in the immunocompromised patient population. We report a bone marrow transplant patient who developed severe abdominal pain accompanied by marked elevations in serum pancreatic and hepatic enzyme levels. She died shortly thereafter. Autopsy revealed hemorrhagic pancreatitis and fulminant hepatic necrosis. Both the pancreas and liver contained intranuclear inclusions consistent with adenovirus; electron microscopy confirmed that there were, indeed, adenoviral particles. This report of adenoviral pancreatitis emphasizes the diversity of manifestations seen with adenoviral infection. PMID- 8406427 TI - Bacterial infections and gastrointestinal lymphomas. PMID- 8406428 TI - The diagnosis of Hirschsprung's disease. PMID- 8406429 TI - Colonic mucosa in diversion colitis. PMID- 8406430 TI - Diagnosis of human genetic disease using recombinant DNA. Fourth edition. AB - Recombinant DNA methodology has greatly increased our knowledge of the molecular pathology of the human genome at the same time as providing the means of diagnosing inherited disease at the DNA level. Direct detection and analysis of a wide range of genetic lesions are now possible using cloned gene or oligonucleotide probes or by direct sequencing of the disease gene(s). In addition, the use of restriction fragment length polymorphisms (RFLPs) within and around these genes as indirect genetic markers has potentiated the tracking of disease alleles in affected pedigrees in cases where direct analysis is not yet feasible. RFLPs associated with linked anonymous DNA segments may also be used not only to diagnose hitherto undetectable disease states, but also for the chromosomal localization of the loci responsible. We present here an update to our previous list of reports describing the direct and indirect analysis/diagnosis of human inherited disease. This compilation is intended to serve as a guide to current molecular genetic approaches in diagnostic medicine. PMID- 8406431 TI - Duplication detection in Japanese Duchenne muscular dystrophy patients and identification of carriers with partial gene deletions using pulsed-field gel electrophoresis. AB - DNA samples from 21 unrelated Japanese patients with Duchenne muscular dystrophy (DMD) with nondeletion-type abnormality in the dystrophin gene and three samples from possible deletion carriers were analyzed using pulsed-field gel electrophoresis (PFGE). Among the 21 patients, 7 were found to carry partial duplications of the dystrophin gene spanning 50-400 kb. Of these 7 patients, 4 carried duplications corresponding to the major hot-spot regions for deletions (7.5-8.5 kb from the 5' end of cDNA), whereas two cases contained duplications in a region about 10 kb from the 5' end of cDNA, where causative mutations are reported to be rare. Only 1 case was found to contain a duplication of a region about 1 kb from the 5' end of cDNA, which is the reported duplication prone region. A combination of Southern blot analyses of conventional agarose gel electrophoresis and PFGE was confirmed to be useful, not only for detecting duplications and deletions, per se, but also for identifying carriers in the affected family. PMID- 8406432 TI - Loss of heterozygosity and K-ras gene mutations in gastric cancer. AB - In order to identify relevant genetic lesions in gastric carcinoma, we searched for tumor suppressor gene inactivation and K-ras gene mutations by analyzing tumor and control DNAs from 34 patients. These were from an epidemiologically defined area of Italy characterized by one of the world's highest incidences of stomach cancer. Allele losses were investigated by the Southern blotting procedure at 16 polymorphic loci on 11 different chromosomes. Our data demonstrate that chromosomal regions 5q, 11p, 17p and 18q are frequently deleted, and that 7q and 13q chromosome arms are also involved, although at a lower frequency. Loss of heterozygosity (LOH) at region 11p was not found during other surveys carried out on patients of different geographic origins. No specific combination of allelic losses could be recognized in the samples analyzed, the only exception being that tumors with 17p allelic loss also showed LOH on the 18q region. When matching frequent LOH events and the stage of progression of the tumors, we observed a trend of association between advanced stages and allelic losses on 17p and 18q chromosome arms. The analysis of K-ras, carried out by the polymerase chain reaction and denaturing gradient gel electrophoresis, demonstrated transforming mutations in only 3 out of 32 cases. Colorectal tumorigenesis proceeds by the accumulation of genetic alterations, including K ras mutations and inactivation of tumor suppressor genes on the 5q, 17p and 18q regions. Our data indicate that, although gastric and colorectal neoplasias share common genetic alterations, they probably progress through different pathways. PMID- 8406433 TI - Human chromosome 11 complements ataxia-telangiectasia cells but does not complement the defect in AT-like Chinese hamster cell mutants. AB - It has been shown that the X-ray-sensitive Chinese hamster V79 mutants (V-E5, V C4 and V-G8) are similar to ataxia-telangiectasia (A-T) cells. To determine whether the AT-like rodent cell mutants are defective in the gene homologous to A T (group A, C or D), human chromosome 11 was introduced to the V-E5 and V-G8 mutant cells by microcell-mediated chromosome transfer. Forty independent hybrid clones were obtained in which the presence of chromosome 11 was determined by in situ hybridization. The presence of the region of chromosome 11q22-23 was shown by molecular analysis using polymorphic DNA markers specific for the ATA, ATC and ATD loci. Seventeen of the obtained monochromosomal Chinese hamster hybrids contained a cytogenetically normal human chromosome 11, but only twelve hybrid cell lines were shown to contain an intact 11q22-23 region. Despite the complementation of the X-ray sensitivity by a normal chromosome 11 introduced to A-T cells (complementation group D), these twelve Chinese hamster hybrid clones showed lack of complementation of X-ray and streptonigrin hypersensitivity. The observed lack of complementation does not seem to be attributable to hypermethylation of the human chromosome 11 in the rodent cell background, since 5-azacytidine treatment had no effect on the streptonigrin hypersensitivity of the hybrid cell lines. These results indicate that the gene defective in the AT like rodent cell mutants is not homologous to the ATA, ATC or ATD genes and that the human gene complementing the defect in the AT-like mutants seems not to be located on human chromosome 11. PMID- 8406434 TI - Retrospective molecular detection of Transthyretin Met 111 mutation in a Danish kindred with familial amyloid cardiomyopathy, using DNA from formalin-fixed and paraffin-embedded tissues. AB - Severe familial amyloid cardiomyopathy (FAC) in a Danish kindred is associated with a specific mutation (Met for Leu 111) in the transthyretin (TTR) gene. The mutation causes the loss of a DdeI restriction site in the gene, allowing molecular diagnostic studies. We studied formalin-fixed, paraffin-embedded tissues, up to 39 years old, from 29 family members of this kindred. DNA was partially purified from deparaffinized tissue sections and a DNA sequence of the TTR gene flanking the mutation site was amplified by the polymerase chain reaction (PCR), followed by restriction enzyme analysis. Amplified DNA was obtained from tissues representing 23 of the 29 persons. Ten out of the 23 family members were found to carry the TTR Met 111 mutation, whereas 13 were not affected. The results were consistent with known clinical data and with corresponding serum TTR examinations. This retrospective study shows that archival tissues can be used to confirm the diagnosis and disease pattern in members of families affected by hereditary diseases. PMID- 8406435 TI - Founder effect in a Belgian-Dutch fragile X population. AB - For many years, the high prevalence of the fragile X syndrome was thought to be caused by a high mutation frequency. The recent isolation of the FMR1 gene and identification of the most prevalent mutation enable a more precise study of the fragile X mutation. As the vast majority of fragile X patients show amplification of an unstable trinucleotide repeat, DNA studies can now trace back the origin of the fragile X mutation. To date, de novo mutations leading to amplification of the CGG repeat have not yet been detected. Recently, linkage disequilibrium was found in the Australian and US populations between the fragile X mutation and adjacent polymorphic markers, suggesting a founder effect of the fragile X mutation. We present here a molecular study of Belgian and Dutch fragile X families. No de novo mutations could be found in 54 of these families. Moreover, we found significant (P < 0.0001) linkage disequilibrium in 68 unrelated fragile X patients between the fragile X mutation and an adjacent polymorphic microsatellite at DXS548. This suggests that a founder effect of the fragile X mutation also exists in the Belgian and Dutch populations. Both the absence of new mutations and the presence of linkage disequilibrium suggest that a few ancestral mutations are responsible for most of the patients with fragile X syndrome. PMID- 8406436 TI - Long range restriction map of the von Hippel-Lindau gene region on human chromosome 3p. AB - Von Hippel-Lindau disease is a heritable tumour syndrome caused by the loss of the function of a tumour suppressor gene on the short arm of human chromosome 3. The interval RAF1-D3S18 (3p25-3p26) has been identified by genetic linkage studies to harbour the von Hippel-Lindau gene. We have constructed a long range restriction map of this region and have succeeded in demonstrating the physical linkage of loci D3S726 (DNA probe LIB31-38), D3S18 (c-LIB-1, L162E5), D3S601 (LIB19-63) and D3S587 (LIB12-48). Since multipoint analysis has located D3S601 proximal to D3S726, the physical map should be oriented with D3S726 towards the telomere. The order and distances of probes within the von Hippel-Lindau gene region is as follows: telomere--LIB31-38--(< 280 kb)--c-LIB-1--(overlapping)- L162E5--(900-1600 kb)--(LIB19-63, LIB12-48)--centromere. In tissues that included blood, semen and Epstein-Barr-virus-transformed lymphocytes, we detected a putative CpG island flanking D3S18. PMID- 8406437 TI - Genetic mapping of the erythropoietin receptor gene. AB - We describe a novel, highly informative (polymorphism information content, PIC, = 0.86) simple sequence repeat polymorphism at the 5' end of the gene encoding the human erythropoietin receptor (EPOR) previously assigned to 19p13.2 by in situ hybridization. Fourteen different allelic size variants were identified in 12 families of the CEPH (Centre d'Etude du Polymorphisme Humain) family panel of 40 families. In pairwise linkage 16 of the 65 chromosome 19 markers reported to the CEPH database gave a lod score exceeding 3.0 when tested against EPOR. The most likely location of EPOR within a framework of 10 markers including orientation and information on reported physical assignments was pter-[INSR-D19S177-D19S176] D19S24-LDLR-++ +EPOR-cen-D19S7-D19S49-D19S75-D19S47-AP OC2-qter, placing EPOR as the most proximal of the tested loci on the short arm. On an 11-point map the position and order for all other loci except INSR were supported by the data with odds exceeding 1,000:1. The polymorphism at the 5' end of EPOR should provide a useful landmark marker for future mapping studies of this region. PMID- 8406438 TI - Localization of X chromosome short arm markers relative to synovial sarcoma- and renal adenocarcinoma-associated translocation breakpoints. AB - A series of thirteen different DNA markers was mapped relative to papillary renal cell carcinoma- and synovial sarcoma-associated translocation breakpoints in Xp11.2 using a panel of tumor-derived somatic cell hybrids in conjunction with Southern blot analysis. Our results indicate that the two translocation breakpoints differ from each other and that the chromosomal break in t(X;1) positive papillary renal cell carcinoma is located between the markers PFC-TIMP OATL1-SYP-TFE3 and DXS226-DXS146-DXS255-OATL2-DXS14. In addition, our current breakpoint analysis has resulted in a revision of the regional localization of the proximal Xp marker DXS226. PMID- 8406439 TI - Fetal cells in the maternal circulation during first trimester in pregnancies. AB - To investigate the presence of fetal cells in the maternal circulation during early pregnancy, the polymerase chain reaction was used to test the presence of human Y chromosome-specific ZFY and SRY gene DNA sequences in maternal peripheral blood specimens from 19 women carrying male fetuses and 12 women carrying female fetuses. The presence of fetal cells was suggested as early as 6 weeks gestation in 1 of the 19 women bearing male fetuses. Fetal cells were present in the maternal circulation of 15 of the 19 women by 9 weeks gestation, and in only 1 of the 19 were fetal cells not detected until the 12th week after conception. These results suggest that identification of fetal cells in the maternal circulation is possible with a properly designed and executed polymerase chain reaction. However, there was considerable variation with respect to when these fetal cells first became detectable during pregnancy. These fetal cells are potentially a valuable source of material for biochemical and genetic studies of the fetuses. PMID- 8406440 TI - A radiation hybrid map of human chromosome 11q22-q23 containing the ataxia telangiectasia disease locus. AB - We describe a high-resolution radiation hybrid map of human chromosome 11q22-q23 containing the ataxia-telangiectasia (AT) disease gene loci. The order and intermarker distances of 32 chromosome 11q22-q23 markers were determined by a multipoint maximum likelihood method of analysis of the cosegregation of markers in 100 radiation hybrids. The radiation hybrid map of polymorphic loci was consistent with genetic linkage maps of common markers. Several genes, including alpha B-crystallin, adrenal ferrodoxin, CBL2, collagenase, dopamine receptor type 2, neural cell adhesion molecule, progesterone receptor, and stromelysins 1 and 2, were placed in relation to previously ordered, genetically mapped polymorphic loci. Five new markers (alpha B-crystallin, adrenal ferrodoxin, CJ52.114, CJ52.3, and D11S535) were ordered within the current published flanking markers for the AT group A and group C disease loci. A candidate AT group D gene (ATDC) identified by Kapp et al. (1992, Am. J. Hum. Genet. 51: 45-54) was mapped telomeric to THY1, outside the flanking markers identified by multipoint linkage analysis for the major AT locus. PMID- 8406441 TI - Comparative mapping of the Grpr locus on the X chromosomes of man and mouse. AB - The gastrin-releasing peptide receptor has been previously cloned from both humans and mice. We have mapped the mouse gastrin-releasing peptide receptor (Grpr) locus using a polymorphic CAn repeat located in the 5' untranslated region of the gene and a Mus spretus/Mus musculus interspecific backcross. The Grpr locus mapped between the Pdha-1 and Amg loci on the mouse X chromosome. Studies in man indicate that GRPR maps to the Xp21.2-p22.3 region of the human X chromosome and not to the Xp11-q11 interval as previously reported. The assignment of the GRPR locus to the distal Xp region is supported by the comparative map position in the mouse. PMID- 8406442 TI - High-resolution comparative mapping of mouse chromosome 17. AB - We have constructed a high-resolution genetic linkage map spanning the proximal 32 cM of mouse chromosome 17 including the t complex. Simple sequence repeats at D17Tu1 and D17Mit6 were employed to identify 121 recombinants among 374 offspring of a (C57BL/6 x CAST/Ei) x C57BL/6 backcross. In contrast to previously reported interspecific Mus domesticus x Mus spretus backcrosses, we did not observe inversion polymorphisms with our cross. This and the relatively high frequency of DNA polymorphisms between C57BL/6 and CAST/Ei allowed us to put 32 RFLV markers and 32 PCR markers on a single map. We present the localization of four new DNA markers and determined map positions for 10 other loci, which previously had been assigned to intervals of the t complex only through the study of partial t haplotype chromosomes. PMID- 8406443 TI - A tetranucleotide repeat mouse minisatellite displaying substantial somatic instability during early preimplantation development. AB - The highly variable mouse minisatellite Hm-2 is located on chromosome 9 and consists of GGCA tetranucleotide repeats with alleles containing up to 5000 repeat units. This locus is unstable with a germline mutation rate to new length alleles of at least 3.6% per gamete. Hm-2 also shows substantial somatic instability, producing mutational mosaicism detectable in 20% of adult mice. Analysis of allele dosage in mice carrying somatic mutations, plus studies of mosaicism in mouse embryos and extraembryonic tissues, suggests that somatic mutant alleles preferentially arise during preimplantation development and particularly during the first two cell divisions after fertilization. PMID- 8406444 TI - The genes for oncostatin M (OSM) and leukemia inhibitory factor (LIF) are tightly linked on human chromosome 22. AB - Oncostatin M (OSM) and leukemia inhibitory factor (LIF) are members of a family of cytokines that regulate the proliferation and differentiation of a variety of cell types. In this report, cDNA probes specific for OSM and LIF were hybridized to DNA from somatic cell hybrids containing defined regions of human chromosome 22, and the gene for human OSM was found to segregate with that of LIF. Southern analysis of high-molecular-weight DNA that had been digested with rare-cutting restriction enzymes and analyzed by pulsed-field gel electrophoresis showed identical hybridization patterns with both probes. The probes also identified common cosmid clones on high-density cosmid filters prepared from chromosome 22 specific flow-sorted cosmid libraries. Restriction and Southern analyses of six cosmid clones established a contig of approximately 100 kb surrounding the genes for OSM and LIF. The OSM and LIF genes are tandemly arranged in the same transcriptional orientation separated by approximately 10 kb. The direction of gene transcription is telomeric to centromeric, with the OSM gene located upstream of the LIF gene. Our studies define a new gene cluster on chromosome 22 and provide strong evidence that OSM and LIF have resulted from duplication of a common ancestral gene. PMID- 8406445 TI - Molecular analysis of phenylketonuria in Denmark: 99% of the mutations detected by denaturing gradient gel electrophoresis. AB - We present the results of a comprehensive study on the molecular basis of phenylketonuria (PKU) in Denmark. A strategy relying on PCR in combination with denaturing gradient gel electrophoresis for analyzing the coding sequence and splice site junctions of the phenylalanine hydroxylase gene allowed us to detect a molecular defect on 99% of 308 Danish PKU chromosomes. The mutational spectrum consists of 35 different mutations, including 23 missense mutations, 5 splice mutations, 4 nonsense mutations, and 3 deletions. Seventeen of these mutations have not been reported previously. The mutation detection assay presented in this report offers a simple and reliable methodological entity that can be applied to rapid diagnosis and carrier detection of phenylketonuria in any population, irrespective of the frequency and distribution of mutations. PMID- 8406446 TI - Physical mapping of DNA markers in the q13-q22 region of the human X chromosome. AB - DNA probe screening of somatic cell hybrids derived from females with X;autosome translocations has enabled us to define eight new breakpoints within the Xq13-q22 region. Together with other X-chromosome rearrangements that have been described earlier, these breakpoints subdivide the Xq21-q22 region into 20 intervals. Our panel refines the physical assignment of 40 probes in the Xq21-q22 segment. Thus, these X-chromosome rearrangements are useful tools for ordering X-linked markers and genes on the proximal long arm of the human X chromosome. PMID- 8406447 TI - High-resolution mapping of probes near the X-linked lymphoproliferative disease (XLP) locus. AB - Fluorescence in situ hybridization (FISH) was employed in high-resolution mapping of probes near the X-linked lymphoproliferative disease (XLP) locus. The map includes the DXS42, DXS12, DXS6, DXS982, DXS739, DXS75, DXS100, DXS10, and DXS177 loci. Metaphase analysis showed that DXS12 and DXS42 mapped to proximal Xq25, while DXS10 and DXS177 mapped to proximal Xq26.1. DXS6, DXS982, DXS739, DXS75, and DXS100 were in Xq25. The order of probes deduced from interphase FISH was: Xq24-(DXS12, DXS42)-DXS6-DXS982-DXS739-DXS75-DXS100+ ++-DXS10-DXS177-Xq26.2. We estimate that the entire region between DXS12 and DXS177 is about 7 Mb. Our previous study indicated that all three XLP deletions (63-3, 66-1, and 43-4) lacked DXS739. We now report that DXS75 and DXS982 are also missing in these deletions. Using interphase FISH measurements, we estimate that 2 Mb are absent in 63-3, and 4 Mb are absent in 66-1 and 43-4. This FISH map confines the XLP candidate gene region to a 2-Mb interval between DXS6 and DXS100 and places DXS100 distal to the XLP locus. This study also demonstrates that small probes (0.6 to 3.6 kb) can be utilized in FISH. PMID- 8406448 TI - C4BPAL1, a member of the human regulator of complement activation (RCA) gene cluster that resulted from the duplication of the gene coding for the alpha-chain of C4b-binding protein. AB - The regulator of complement activation (RCA) gene cluster evolved by multiple gene duplications to produce a family of genes coding for proteins that collectively control the activation of the complement system. We report here the characterization of C4BPAL1, a member of the human RCA gene cluster that arose from the duplication of the C4BPA gene after the separation of the rodent and primate lineages. C4BPAL1 maps 20 kb downstream of the C4BPA gene and is the same 5' to 3' orientation found for all RCA genes characterized thus far. It includes nine exon-like regions homologous to exons 2-8, 11, and 12 of the C4BPA gene. Analysis of the C4BPAL1 sequence suggests that it is currently a pseudogene in humans. However, comparisons between C4BPAL1 and the human and murine C4BPA genes show sequence conservation, which strongly suggests that, for a long period of time, C4BPAL1 has been a functional gene coding for a protein with structural requirements similar to those of the alpha-chain of C4b-binding protein. PMID- 8406449 TI - Nucleotide sequence, expression, and chromosomal mapping of Mrp and mapping of five related sequences. AB - We isolated and characterized a genomic clone for the mouse MARCKS-related protein, or MRP, also known as F52 and MacMARCKS. A 3699-bp plasmid contained 407 bp of 5'-flanking region, 186 bp of 5'-untranslated region, 600 bp of protein coding region, 784 bp of a single intron, 746 bp of 3'-untranslated region, and 976 bp of 3'-flanking region. The position of the single intron was identical to the intron position in all known MARCKS mRNAs. When the plasmid containing the genomic sequences was transfected into fibroblasts lacking endogenous Mrp expression, the 407 bp of promoter conferred high-level expression of the full length, spliced mRNA. The putative promoter was therefore functional; however, despite tissue-specific regulation and transcriptional induction in some cells in a manner similar to that seen with MARCKS expression, the promoters were highly dissimilar at the level of primary sequence (37% identity over 407 bp). Mrp mapped to a position on mouse chromosome 4 that was closely linked to the Lck locus. Numerous additional species that hybridized to the MRP cDNA were noted on Southern blotting of mouse genomic DNA. Five related loci were labeled Mrp-rs1 through Mrp-rs5 (for Mrp-related sequences) and were mapped to mouse chromosomes 10, 17, 15, 13, and X, respectively. Three of these related sequences have been cloned, and all appear to represent pseudogenes. PMID- 8406450 TI - Multipoint mapping of the central core disease locus. AB - A linkage analysis with 12 DNA markers from proximal 19q was performed in eight families with central core disease (CCO). Two-point analysis gave a peak lod score of Z = 4.95 at theta = 0.00 for the anonymous marker D19S190 and of Z = 2.53 at theta = 0.00 for the ryanodine receptor (RYR1) candidate gene. Multipoint linkage data place the CCO locus at 19q13.1, flanked proximally by D19S191/D19S28 and distally by D19S47. This map location includes the RYR1 gene. The results of the linkage study present no evidence for genetic heterogeneity of CCO. PMID- 8406451 TI - Isolation and characterization of three microsatellite markers in the proximal long arm of the human X chromosome. AB - Three microsatellites have been identified in cosmids from the human X chromosome. The cosmids have been assigned locus numbers DXS554, DXS559, and DXS566 and have been localized to Xq12-q13 (DXS554 and DXS559) and Xq13 (DXS566). In addition, they have been genetically mapped in relation to the androgen receptor (AR), phosphoglycerate kinase 1, pseudogene 1 (PGK1P1), and phosphoglycerate kinase (PGK1) loci in the proximal long arm. Genetically, the localization of microsatellites at DXS554 and DXS566 is indistinguishable from PGK1, whereas that at DXS559 maps between AR and PGK1, close to PGK1P1. DXS566 is identical to the independently identified DXS441 marker. These markers should be useful for physical and genetic mapping in this region. PMID- 8406452 TI - Fifty sequenced-tagged sites on human chromosome 11. AB - Fifty novel sequenced-tagged sites (STSs) were identified from cosmid clones mapped to human chromosome 11. DNA sequences were determined for one or both cloning ends of 69 cosmid markers that had each been localized to 1 of 24 subchromosomal regions by means of hybridization to somatic cell hybrid panels. Proper primer sequences and appropriate conditions for a polymerase chain reaction (PCR) were determined for each marker. Twenty-one of the cosmids were not suitable for generating STSs, mainly because both of their ends contained repetitive elements such as Alu and L1 sequences; however, some were inappropriate because the sizes of their PCR products from human DNA, used as template, were same as those from yeast DNA. Finally, 50 STSs were established from 48 clones: 20 were derived from markers localized on the short arm and 30 from the long arm. These STSs can serve as new reagents for investigating human DNA in somatic cell hybrids and for isolating yeast artificial chromosomes to anchor large DNA contigs and fine-scale physical maps of chromosome 11. PMID- 8406453 TI - A homoallelic Gly317-->Asp mutation in ALPL causes the perinatal (lethal) form of hypophosphatasia in Canadian mennonites. AB - We have discovered a single homoallelic nucleotide substitution as the putative cause of the perinatal (lethal) form of hypophosphatasia in Canadian Mennonites. Previous linkage and haplotype analysis in this population suggested that a single mutational event was responsible for this autosomal recessive form of hypophosphatasia. The mutation is a guanosine-to-adenosine substitution at nucleotide position 1177 in exon 10 of the tissue nonspecific (liver/bone/kidney) alkaline phosphatase gene. This Gly317-->Asp mutation segregates exclusively with the heterozygote phenotype we previously assigned by biochemical testing (maximum combined lod score of 18.24 at theta = 0.00). This putative disease-causing mutation has not been described in controls nor in other non-Mennonite probands with both lethal and nonlethal forms of hypophosphatasia studied to date. This Gly317-->Asp mutation changes a polar glycine to an acidic aspartate at amino acid position 317 within the highly conserved active site region of the 507-amino acid polypeptide. Carrier screening for this lethal mutation in our high-risk population is now feasible. PMID- 8406454 TI - A fourth example suggests that premature termination codons in the COL2A1 gene are a common cause of the Stickler syndrome: analysis of the COL2A1 gene by denaturing gradient gel electrophoresis. AB - A series of oligonucleotide primers was designed to generate polymerase chain reaction products that contained exons 6 to 49 of the human gene for type II procollagen (COL2A1) and that could be used to detect sequence variations by denaturing gradient gel electrophoresis. To improve the sensitivity of the analysis, GC clamps were introduced into one primer of each pair. The procedure successfully detected 10 neutral single-base variations in the gene. In addition, the procedure detected a single-base deletion in exon 43 that introduced a premature termination codon in exon 44 and caused the Stickler syndrome (arthro ophthalmopathy) in one family. The mutation is the fourth mutation in the COL2A1 gene shown to cause the Stickler syndrome. The mutation is similar to the first three mutations causing the disease in that they also introduced premature termination signals. Since only one mutation introducing a premature termination codon was found in the course of defining 120 or more mutations in type I and III procollagens, the results suggest that such mutations may have a special relationship to the Stickler syndrome. PMID- 8406455 TI - Assignment of the nuclear mitotic apparatus protein NuMA gene to human chromosome 11q13. AB - A monoclonal antibody that was specific for a nuclear matrix protein was obtained and used to screen a human lambda gt11 expression library. Several partial cDNA clones were isolated and sequenced. The sequence for this protein was shown to be identical to that of NuMA, a 236-kDa nuclear mitotic spindle apparatus protein. NuMA has been recently characterized by two independent studies, and is thought to be part of a family of proteins that is required for the completion of mitosis. In this report, the chromosomal localization and copy number of the NuMA gene are analyzed using cDNA clones. High-resolution in situ hybridization reveals a single pair of signals on sister chromatids of human chromosome 11 at band q13. Stringent Southern analysis of human genomic DNA resulted in simple restriction patterns. These results together indicate that the NuMA gene is present as a single copy on human chromosome 11q13. PMID- 8406456 TI - Sequence and characterization of the complete human thrombospondin 2 cDNA: potential regulatory role for the 3' untranslated region. AB - We report the remainder of the cDNA sequence for human thrombospondin 2 (hTSP2) including the 5' and 3' untranslated regions. The homology between the two human thrombospondins is highest at the carboxyl-terminal end. The 5' untranslated region of hTSP2 is a GC-rich, 239-bp segment that encodes an in-frame methionine residue embedded in a Kozak consensus sequence. The 3' untranslated region is AT rich and contains nucleotide sequences implicated in the regulation of mRNA stability. It also contains an 89-bp AT-rich segment with 92% of its nucleotides identical to the same region of the mouse TSP2 3' untranslated region, which suggests an important functional or structural property associated with this region. PMID- 8406457 TI - Molecular analysis and genetic mapping of the rhodopsin gene in families with autosomal dominant retinitis pigmentosa. AB - Eighty-eight patients/families with autosomal dominant retinitis pigmentosa (RP) were screened for rhodopsin mutations. Direct sequencing revealed 13 different mutations in a total of 14 (i.e., 16%) unrelated patients. Five of these mutations (T4K, Q28H, R135G, F220C, and C222R) have not been reported so far. In addition, multipoint linkage analysis was performed on two large families with autosomal dominant RP due to rhodopsin mutations by using five DNA probes from 3q21-q24. No tight linkage was found between the rhodopsin locus (RHO) and D3S47 (theta max = 0.08). By six-point analysis, RHO was localized in the region between D3S21 and D3S47, with a maximum lod score of 13.447 directly at D3S20. PMID- 8406458 TI - A high-resolution cytogenetic map of human chromosome 5: localization of 206 new cosmid markers by direct R-banding fluorescence in situ hybridization. AB - We have constructed a high-resolution cytogenetic map of human chromosome 5 with 206 new cosmid clones by direct R-banding fluorescence in situ hybridization. The fluorescent signals of 206 clones were evenly distributed throughout chromosome 5, although they were sublocalized preferentially to R-positive bands. This high resolution cytogenetic map with an average distance of nearly 1 Mb will serve as important resource for construction of a genetic linkage map, which is essential for positional cloning of responsible genes. Moreover, these mapping data provide many useful landmarks for the construction of contig maps of chromosome 5, as required in the Human Genome Project. PMID- 8406459 TI - Mapping of the human NMDA receptor subunit (NMDAR1) and the proposed NMDA receptor glutamate-binding subunit (NMDARA1) to chromosomes 9q34.3 and chromosome 8, respectively. AB - A role for the N-methyl-D-aspartate (NMDA) receptor in the molecular pathology underlying Huntington disease (HD) has been proposed on the basis of neurochemical studies in HD and the ability of the NMDA receptor to mediate neuronal cell death. The molecular cloning of the human NMDA receptor subunit (NMDAR1) and a proposed glutamate-binding subunit of the NMDA receptor (NMDARA1) have provided an opportunity to test the hypothesis that either of these genes might be directly involved in the causation of HD. We have mapped NMDAR1 to 9q34.3 using in situ hybridization studies and NMDARA1 to human chromosome 8 using a somatic cell hybrid panel. Because the gene causing HD has been localized to chromosome 4p16.3, the chromosome assignments reported here are inconsistent with either of these genes playing a causative role in the molecular pathology of HD. However, it is noteworthy that the gene for torsion dystonia has also been localized by genetic studies to 9q34.3, the same regional map location as NMDAR1. PMID- 8406460 TI - Mapping of the human CENP-B gene to chromosome 20 and the CENP-C gene to chromosome 12 by a rapid cycle DNA amplification procedure. AB - By optimizing the primer-annealing temperature in a rapid air cycling procedure, two human DNA sequences encoding centromere proteins B and C (CENP-B and CENP-C) were specifically amplified without any detectable amplification of highly homologous rodent DNA sequences. Using a panel of rodent/human hybrid DNA, the genes for human CENP-B and CENP-C were conveniently mapped to chromosomes 20 and 12, respectively. PMID- 8406461 TI - Molecular characterization of an intragenic minisatellite (VNTR) polymorphism in the human parathyroid hormone-related peptide gene in chromosome region 12p12.1 p11.2. AB - The human parathyroid hormone-related peptide (hPTHrP) gene in chromosome region 12p12.1-p11.2 plays an important role in mammalian development and specifically in skeletogenesis. We have characterized a VNTR polymorphism in the hPTHrP gene that is located in an intron 100-bp downstream of exon VI that encodes a 3' untranslated region. By PCR analysis eight different alleles were identified in a group of 112 unrelated individuals. All eight alleles were sequenced and the repeat unit was identified as the general sequence [G(TA)nC]N, where n = 4 to 11 and N = 3 to 17. This polymorphic sequence-tagged site will be useful for mapping chromosome 12p and will aid in testing for linkage of genetic diseases to the hPTHrP gene. PMID- 8406462 TI - A continuous linkage map of 22 short tandem repeat polymorphisms on human chromosome 12. AB - A continuous linkage map consisting of 22 short tandem repeat polymorphisms has been constructed for human chromosome 12 using 23 non-CEPH pedigrees. The markers were distributed at an average distance of 9.35 cM (3.1-33.9 cM). Eighteen of the markers could be positioned uniquely with a likelihood of > 1000:1. The physical locations of some of the markers suggest that the map covers 85-95% of the chromosome. This framework map of 18 markers has a female length of 213 cM and a male length of 131 cM. Female recombination frequencies were greater than male recombination frequencies except in the distal portion of the short arm. The map provides confirmatory evidence for orders established previously on CEPH pedigrees and uniquely positions 4 additional markers (CD4, ATPSB, D12S56, PLA2). PMID- 8406463 TI - Physical analysis of the terminal 240 kb of DNA from human chromosome 7q. AB - DNA from a 240-kb human telomeric yeast artificial chromosome (HTY) clone was analyzed using physical mapping methods. Cosmid subclones of the YAC were fingerprinted using restriction enzyme digestion and human repeat sequence hybridization and then assembled into two contigs that together span 93% of the human insert. Data from restriction mapping and Bal31 exonuclease experiments indicate that, except for the truncation of distal genomic (T2AG3)n sequences, the molecular clone HTY 146 contains a contiguous, 230-kb telomere-terminal fragment from 7qter. Markers derived from this clone will allow telomeric closure of the physical and genetic linkage maps of human chromosome 7q. PMID- 8406464 TI - Assignment of the chromogranin A (Chga) locus to homologous regions on mouse chromosome 12 and rat chromosome 6. AB - Chromogranin A is an acidic protein, stored and released with catecholamines, and is overexpressed in genetic hypertension. In the human genome, its locus has previously been positioned on the long arm of Chromosome 14 in the 14q32 region. As a first step toward evaluating its potential linkage with hereditary hypertension, we determined its chromosomal position in mouse and rat. Chromogranin A was present as a single-copy gene in both mouse and rat. Analysis of the allele distribution in an interspecific mouse backcross by single-strand conformation polymorphism positioned the chromogranin A locus on Chromosome 12, between Igh-C and D12Pas1. Evaluation of a rat/mouse somatic cell hybrid panel indicated that chromogranin A is on rat Chromosome 6. In each case (mouse, rat, and human), chromogranin A is in a conserved region with nearby markers including the immunoglobulin heavy chain locus. PMID- 8406465 TI - D21S418E identifies a cAMP-regulated gene located on chromosome 21q22.3 that is expressed in placental syncytiotrophoblast and choriocarcinoma cells. AB - A partial cDNA (D21S418E) whose nucleotide sequence has no significant homologies with known mammalian DNA sequences was isolated from a human placental library. The cDNA hybridized with a 10-kb transcript present in term placenta. Messages of 10 and 7.5 kb were induced in BeWo and JEG-3 choriocarcinoma cells by treatment with 8-Br-cAMP. The mRNA was not detected in human brain, liver, lung, kidney, pancreas, heart, skeletal muscle, or myometrium. The D21S418E locus was assigned to a 3.5-Mb region of chromosome 21q22.3. PMID- 8406466 TI - Demonstration of physical linkage between the promoter region and the polymorphic kringle IV domain in the Apo(a) gene by pulsed-field gel electrophoresis. PMID- 8406467 TI - Localization of the human indoleamine 2,3-dioxygenase (IDO) gene to the pericentromeric region of human chromosome 8. PMID- 8406468 TI - Localization of the estrogen receptor locus (ESR) to chromosome 6q25.1 by FISH and a simple post-FISH banding technique. PMID- 8406469 TI - Leukocyte common antigen-related phosphatase (LRP) gene structure: conservation of the genomic organization of transmembrane protein tyrosine phosphatases. AB - The leukocyte common antigen-related protein tyrosine phosphatase (LRP) is a widely expressed transmembrane glycoprotein thought to be involved in cell growth and differentiation. Similar to most other transmembrane protein tyrosine phosphatases, LRP contains two tandem cytoplasmic phosphatase domains. To understand further the regulation and evolution of LRP, we have isolated and characterized mouse lambda genomic clones. Thirteen genomic clones could be divided into two non-overlapping clusters. The first cluster contained the transcription initiation site and the exon encoding most of the 5' untranslated region. The second cluster contained the remaining exons encoding the protein and the 3' untranslated region. The gene consists of 22 exons spanning over 75 kb. The distance between exon 1 and exon 2 is at least 25 kb. Characterization of the 5' ends of LRP mRNA by S1 nuclease protection identifies putative initiation start sites within a G/C-rich region. The upstream region does not contain a TATA box. Comparison of the LRP gene structure to the mammalian protein tyrosine phosphatase gene, CD45, shows striking similarities in size and genomic organization. PMID- 8406470 TI - Sequence-tagged NotI sites of human chromosome 21: sequence analysis and mapping. AB - We isolated and analyzed 19 NotI site-containing clones specific for human chromosome 21. The overall process consisted of selective isolation of NotI site containing clones from flow-sorted chromosome 21 libraries, selection of independent clones by their restriction patterns and nucleotide sequences, and assignment of the clones to chromosome 21. Sequence analysis showed that the regions around the NotI sites had features typical of CpG islands, such as extremely high GC content, high-frequency appearance of CpG dinucleotide sequences, and lack of methylation of these CpGs. PCR conditions for these extremely G + C-rich templates were optimized to establish these NotI sites as sequence-tagged sites. PMID- 8406471 TI - YAC clone contigs surrounding the Zfx and Pola loci on the mouse X chromosome. AB - Pulsed-field mapping of a number of DNA markers in the Pola-Zfx region of the mouse X chromosome has established a genomic restriction map extending over 1.4 Mb. A number of YAC clones from the Pola-Zfx region have been isolated from three mouse YAC libraries--first, a mouse C57BL/10 partial R1 YAC library constructed in a yeast strain carrying a rad52 mutation (Chartier et al. (1992) Nature Genetics 1: 132-136); second, a mouse C3H partial R1 library (Larin et al. (1991) Proc. Natl. Acad. Sci. USA 88: 4123-4127); and third, a mouse C57BL/6 partial R1 library (Burke et al. (1991) Mamm. Genome 1:65). Six YAC clones encompass the Zfx Pola region, confirming the linkage of the Pola and Zfx loci and establishing a physical map order in this region of cen-Pola-DXCrc140-DXCrc57-Zfx-tel. The close linkage of Pola and Zfx in the mouse genome suggests that the POLA and ZFX loci must also be closely linked on the human X chromosome. PMID- 8406472 TI - Characterization and sequence analysis of rat serine:pyruvate/alanine:glyoxylate aminotransferase gene. AB - We have reported the isolation of genomic clones encoding serine:pyruvate aminotransferase (SPT; also named alanine:glyoxylate aminotransferase, AGT) (T. Oda, T. Funai, and A. Ichiyama, 1990, J. Biol. Chem. 265: 7513-7519). These clones contained the entire SPT/AGT gene of 10 kb. In this work, we characterized this gene. The SPT/AGT gene consists of 11 exons, and the exon-intron boundaries have typical splice donor and acceptor sequences. Determination of the nucleotide sequence up to -1.25 kb from the transcription initiation site revealed the presence of many putative cis elements, some of which may explain the transcriptional regulation of the SPT/AGT gene by glucagon and glucocorticoid. The nucleotide sequence around the 5' flanking region of the rat SPT/AGT gene and the whole gene organization were compared with those of the human SPT/AGT gene. No obvious similarities were observed in the 5' flanking region up to -1.25 kb from the initiation site of the gene, but exons 2 to 10 of the rat and human genes have identical sizes and show high similarities. PMID- 8406473 TI - Molecular cloning and characterization of a human carboxylesterase gene. AB - A cDNA encoding human liver carboxylesterase and its gene were isolated. Nucleotide sequence analyses of the cDNA revealed that the predicted enzyme protein consists of 567 amino acids, including 18 amino acids of a putative signal peptide. Comparison of the deduced amino acid sequences of this enzyme with those of seven other carboxylesterases in various mammalian species, together with experimental data from several other laboratories, showed that these enzymes can be classified into three groups depending on the sequences at their carboxyl terminals and the presence or absence of one exon. A human carboxylesterase gene was found to span approximately 30 kb and to have 14 small exons. Alignments of this gene with those of human cholinesterase and rat cholesterol esterase indicated insertional sites at some introns and homologous amino acid sequences around them, although these genes have different numbers of exons. Thus the results supported the conclusion that these esterases evolved from a common ancestral gene. PMID- 8406474 TI - Molecular basis and consequences of a deletion in the amelogenin gene, analyzed by capture PCR. AB - A mutation that disrupts the gene for one of the major proteins in tooth enamel has been investigated. The mutation is located in the amelogenin gene and causes X-linked amelogenesis imperfecta, characterized by defective mineralization of tooth enamel. We have isolated the breakpoints of a 5-kb deletion in the amelogenin gene on the basis of nucleotide sequence information located upstream of the lesion, using a technique termed capture PCR. The deletion removes five of the seven exons, spanning from the second intron to the last exon. Only the first two codons for the mature protein remain, consistent with the relatively severe phenotype of affected individuals in the present family. The mutation appears to have arisen as an illegitimate recombination event since of 11 nucleotide positions immediately surrounding the two breakpoints, 9 are identical. PMID- 8406475 TI - Mapping of glutamic acid decarboxylase (GAD) genes. AB - Glutamic acid decarboxylase (GAD) catalyzes the synthesis of gamma-aminobutyric acid (GABA), which is known as a major inhibitory neurotransmitter in the central nervous system (CNS), but is also present outside the CNS. Recent studies showed that GAD is the major target of autoantibodies associated with the development of insulin-dependent diabetes mellitus and of the rare stiff man syndrome. Studies of GAD expression have demonstrated multiple transcripts, suggesting several isoforms of GAD. In this study, three different genes were mapped by in situ hybridization to both human and mouse chromosomes. The GAD1 gene was mapped to human chromosome 2q31 and to mouse chromosome 2D in a known region of conservation between human and mouse. GAD2, previously mapped to human chromosome 10p11.2-p12, was mapped to mouse chromosome 2A2-B, which identifies a new region of conservation between human and mouse chromosomes. A potential GAD3 transcript was mapped to human chromosome 22q13 and to mouse chromosome 15E in a known region of conservation between human and mouse. It is concluded that the GAD genes may form a family with as many as three related members. PMID- 8406476 TI - A fluorescence in situ hybridization map of human chromosome 21 consisting of 30 genetic and physical markers on the chromosome: localization of 137 additional YAC and cosmid clones with respect to this map. AB - A fluorescence in situ hybridization (FISH) map of human chromosome 21 was compiled using yeast artificial chromosome (YAC) DNA probes that encode 28 markers physically and/or genetically mapped on the chromosome. Probes that recognize the centromere and rDNA repeat sequences in the p arm were also placed as reference markers on the FISH map. For each probe, the location of the fluorescence hybridization signal was measured on metaphase chromosomes with respect to fractional chromosome length (FL) from p-ter. The location of the markers was established with a standard error of +/- 1.9 Mb using from 9 to 63 FL measurements for each probe. The relative order and separation of the markers as determined by FISH are shown to correspond well to those of other maps of the chromosome. Fifty-one additional YAC and 86 cosmid clones were also localized by FISH with respect to the 30 markers on the chromosome. The cosmids, chosen at random from a flow-sorter chromosome 21 cosmid library, show some biases in chromosome distribution. PMID- 8406477 TI - Extrachromosomal maintenance and amplification of yeast artificial chromosome DNA in mouse cells. AB - Linear and circular forms of a 660-kb yeast artificial chromosome (YAC) containing the human hypoxanthine phosphoribosyltransferase gene were introduced into mouse L A-9 cells by fusion with yeast spheroplasts, and in situ hybridization was used to determine the location and fate of the yeast- and YAC derived DNA in 25 fusion cell lines. Human and yeast DNAs were observed as extrachromosomal DNA molecules, present at least 27 cell divisions after fusion, in half the cell lines. The extrachromosomal molecules replicate extrachromosomally but segregate poorly like plasmids that contain an autonomously replicating sequence element in yeast. This system may allow analysis of DNA requirements for replication and segregation in mammalian cells. The extrachromosomal elements resemble double minutes (DMs), and in five cell lines human and yeast DNAs were present as very large integrated blocks of DNA resembling homogeneously staining regions (HSRs). Thus, the fusion cell lines contain the two characteristic forms of amplified DNA observed in cancer cell lines, DMs and HSRs, indicating that these can be formed from fragments of DNA introduced by cell fusion. PMID- 8406478 TI - Genetic mapping and evaluation of candidate genes for spasmodic, a neurological mouse mutation with abnormal startle response. AB - Spasmodic (spd) is a recessive mouse mutation characterized by a prolonged righting reflex, fine motor tremor, leg clasping, and stiffness. Using an intersubspecific backcross that segregates spd, we placed spd on Chr 11 with the following gene order: Adra-1-3.8 +/- 2.1 cM-Pad-1-6.3 +/- 2.7-(spd, Anx-6, Csfgm, Glr-1, Il-3, Il-4, Il-5, Sparc)-9.1 +/- 2.4-D11 Mit5-2.2 +/- 1.5-Asgr-1. This localization eliminated the alpha 1-adrenergic receptor (Adra-1) and the alpha 1 and gamma 2 subunits of the GABAA receptor as candidate genes. Two other promising candidate genes, annexin VI (Anx-6) and a glutamate receptor (Glr-1), were mapped to within 2.1 cM of the spd locus. Although no recombination was observed between spd and Anx-6 or Glr-1, no evidence was obtained for a lesion in either gene. The presence of normal Anx-6 and Glr-1 mRNA transcripts was confirmed by Northern blot analysis, in situ hybridization, and DNA sequence analysis. The localization of Anx-6 and Glr-1 extends the known synteny homology between human chromosome 5q21-q31 and mouse Chr 11 and reveals the probable chromosomal location of the human counterpart to spd. Synteny homology and phenotypic similarities suggest that spasmodic mice may be a genetic model for the inherited human startle disease, hyperekplexia (STHE). PMID- 8406479 TI - Cloning of a portion of the chromosomal gene and cDNA for human beta-fodrin, the nonerythroid form of beta-spectrin. AB - A 96-bp synthetic oligonucleotide corresponding to an amino acid sequence near the N-terminus of erythroid beta-spectrin was used to screen a human genomic library, and two overlapping recombinants were isolated. DNA sequence analysis established that the genomic fragment encoded beta-fodrin, the nonerythroid form of beta-spectrin, by correlation to a known amino acid sequence of human brain beta-fodrin. The genomic DNA contained regions that cross-hybridized with an erythroid beta-spectrin cDNA probe, and the DNA sequence of these regions revealed a high degree of identity with that of erythroid beta-spectrin and a similar exon/intron organization. A single-copy DNA fragment of the beta-fodrin genomic clone was used to screen a lymphoid cell cDNA library and two recombinants were isolated. The composite DNA sequence of these various genomic and cDNA clones encoded almost all of the first twelve 106 amino acid repeat segments of beta-fodrin that shared 58% identity and 75.5% similarity with the amino acid sequence of beta-spectrin and 66% identity with the nucleotide sequence of beta-spectrin cDNA. The chromosomal localization of the gene was determined to be chromosome 2 by hybridization of a single-copy probe derived from the cloned genomic DNA to DNA of a panel of somatic hybrid cell lines, and in situ hybridization localized the gene to band 2p21. beta-Fodrin was assigned the gene symbol SPTBN1. PMID- 8406480 TI - Assignment of the human pulmonary surfactant protein D gene (SFTP4) to 10q22-q23 close to the surfactant protein A gene cluster. AB - Pulmonary surfactant consists of a complex mixture of phospholipids and several proteins essential to normal respiratory function. Two of the surfactant proteins, SP-A and SP-D, appear to have lectin-like activity relevant to the local phagocytic defense. Using polymerase chain reaction (PCR)-based somatic cell hybrid mapping, the human SP-D gene (SFTP4) was assigned to chromosome 10. A regional mapping panel was assembled and characterized using sequence tagged sites for five loci previously mapped to 10q. SFTP4, the SP-A gene (SFTP1), and the microsatellite D10S109 were placed in the interval 10q22-q23. Low-stringency PCR using the SFTP1 primer pair suggested the presence of at least two additional SP-A-related genes in the same region. With the locus for mannose-binding lectin (MBL) at 10q21, this may be indicative of this region's central role in the evolutionary history of carbohydrate-binding proteins containing collagen-like regions. PMID- 8406481 TI - Human Jk recombination signal binding protein gene (IGKJRB): comparison with its mouse homologue. AB - The mouse Igkjrb protein specifically binds to the immunoglobulin Jk recombination signal sequence. The IGKJRB gene is highly conserved among many species such as human, Xenopus, and Drosophila. Using cDNA fragments of the mouse Igkjrb gene, we isolated its human counterpart, IGKJRB. The human genome contains one functional IGKJRB gene and two types of processed pseudogenes. In situ chromosome hybridization analysis demonstrated that the functional gene is localized at chromosome 3q25, and the pseudogenes (IGKJRBP1 and IGKJRBP2, respectively) are located at chromosomes 9p13 and 9q13. The functional gene is composed of 13 exons spanning at least 67 kb. Three types of cDNA with different 5' sequences were isolated by rapid amplification of cDNA ends, suggesting, the presence of three proteins. The aPCR-1 protein, which possessed the exon 1 sequence, was the counterpart of the mouse RBP-2 type protein. The aPCR-2 and 3 proteins may be specific to human cells because the mouse counterparts were not detected. The amino acid sequences of the human and mouse IGKJRB genes were 98% homologous in exons 2-11, whereas the homology of the human and mouse exon 1 sequences was 75%. PMID- 8406482 TI - Organization of the multiple polymorphic sites of the D19S11 locus within a 650 kb cosmid contig. AB - The D19S11 locus has been previously described as consisting of a complex set of six nonallelic polymorphic sites detected with a combination of four restriction enzymes and three probes that were subcloned from a single cosmid. These probes also hybridized to additional nonvariant fragments on Southern blots of human genomic DNA. In the course of establishing a contig map of human chromosome 19, a set of cosmids that were positive for at least one of the probes defining this locus was identified. These cosmids, along with additional cosmids, were assembled using a combination of strategies, including fluorescence in situ hybridization studies using G1 interphase nuclei and sperm pronuclei as chromatin targets, into a single overlapping set of cosmids that spans approximately 650 kb. Cosmids that are positive for the MEL gene probe are localized at the centromeric end of the spanning path, with some cosmids being positive for both the MEL gene probe and one of the D19S11 probes. The EcoRI fragments with homology to the various probes have been identified; some cosmids have homology to all three D19S11 probes. The positions for five of the six polymorphic sites were localized within a 40-kb region, with four sites within 15 kb. PMID- 8406483 TI - A cosmid and yeast artificial chromosome contig containing the complete ryanodine receptor (RYR1) gene. AB - The ryanodine receptor (RYR1) gene is responsible for some forms of malignant hyperthermia and has been localized to 19q13.1. Central core disease is a genetic myopathy that is genetically linked to RYR1. We have identified an overlapping set of cosmid and YAC clones that spans more than 800 kb and includes the RYR1 gene (approximately 205 kb). Cosmids from this region were identified by screening three chromosome 19 cosmid libraries (11-fold coverage) with six subclones representing the entire RYR1 cDNA. Genomic sequences from positive cosmids were then used as probes to identify additional cosmids. A minimally overlapping set of 23 cosmids was assembled into two contigs on the basis of restriction fragment analysis and hybridization data. Three YAC clones were isolated by screening a human YAC library with selected cosmid inserts. Overlaps among these YACs and the cosmid contigs were determined by hybridizing YAC Alu PCR products to cosmid DNAs. The YACs bridged the gap between the cosmid contigs and extended the contig on both sides. Fluorescence in situ hybridization experiments positioned the RYR1 contig between GPI, MAG, and D19S191 on the proximal side and D19S190, CYP2A, CYP2F, SNRPA, BCKDHA, and other markers on the distal side. The 800-kb contig of cloned reagents will facilitate the detailed characterization of the RYR1 gene and other loci that may be closely related to central core disease. PMID- 8406484 TI - Determination of the specificity of aphidicolin-induced breakage of the human 3p14.2 fragile site. AB - The constitutive 3p14.2 fragile site is the most highly inducible fragile site in the human genome. This locus may predispose chromosome 3 to specific losses due to deletions and translocations that have been associated with several malignancies, including hereditary renal cell carcinoma. Using aphidicolin concentrations of 0.4 and 4.0 microM, we have generated and isolated 23 and 22 respective somatic cell hybrids that contain chromosome 3 short-arm breaks. The breakpoints in these hybrids have been localized with numerous chromosome 3 markers. We have observed that at the low aphidicolin dose, chromosome-3 breaks cluster within the 3p14.2 region, whereas at the high aphidicolin dose, two new loci, one within 3p14.1 and the other near the centromere, become predominantly affected. Our studies have failed to differentiate any of the 3p14.2 breakpoints from each other or from the t(3;8)(p14.2;q24.13) familial renal cell carcinoma translocation breakpoint, suggesting that the fragile site may have played a role in the generation of this balanced translocation. The resulting somatic cell hybrids generated from this work have refined the marker localizations within 3p13-p21.1 and should facilitate the physical characterization of this interesting region. PMID- 8406485 TI - The isolation and characterization of the bovine cytochrome b5 gene, and a transcribed pseudogene. AB - This is the first isolation and characterization of a cytochrome b5(b5) gene. The bovine b5 gene is quite large, spanning about 28 kb and contains six exons. One of these exons appears to code for a reticulocyte-specific sequence similar to that described for human and rabbit b5. All of the splicing junctions conform to the GT-AG consensus rule. The 5' flanking sequence has no obvious TATA box, two CAAT boxes, and contains several G:C-rich SpI motifs indicative of a house keeping gene. In reticulocyte mRNA we found evidence for a transcribed b5 pseudogene, but could not detect sequences coding for the soluble form of b5. We conclude that the soluble form of b5 is derived from the membrane-bound b5 by a post-translational mechanism. PMID- 8406486 TI - Analysis of the Pax-3 gene in the mouse mutant splotch. AB - In a linkage analysis of Pax-3 and splotch no recombinations were found in 117 backcross mice. Molecular analysis of Pax-3 in three alleles of splotch shows a number of significant alterations to the Pax-3 gene. In Sp/Sp embryos, cDNA PCR analysis reveals a shortened transcript in which exon 4 of Pax-3 is deleted due to mutation of the splice acceptor site of intron 3. In the Sp4H allele, the Pax 3 gene is deleted and in Spd embryos, Pax-3 expression is significantly lower than that in normal littermate embryos. The linkage analysis, shortened Pax-3 transcript in Sp, and deletion of Pax-3 in Sp4H described here, together with the previous report of an intragenic deletion in Pax-3 in Sp2H mice and the deletion of Pax-3 in Spr mice, provide strong evidence for the allelic identity of Pax-3 and Sp. PMID- 8406487 TI - The splotch-delayed (Spd) mouse mutant carries a point mutation within the paired box of the Pax-3 gene. AB - The splotch (Sp) mouse mutant displays defects in neural crest cell migration and neural tube closure and serves as a model for the study of spina bifida, exencephaly, and Waardenburg syndrome type I in humans. Recently, we have described alterations in the Pax-3 gene for the radiation-induced Spr and Sp2H alleles and for the original, spontaneously arising Sp allele. Another allele that arose spontaneously at the Sp locus, termed splotch-delayed (Spd), shows a less severe phenotype than the other Sp alleles, including the delayed death of homozygous embryos. To determine the molecular basis underlying this unique phenotype, we have analyzed the integrity of the Pax-3 gene in Spd mutant embryos. Nucleotide sequence analysis of cDNA and genomic clones revealed a G to C transversion at nucleotide 421 of the Pax-3 mRNA transcript, which results in a Gly to Arg substitution at position 42 of the Pax-3 protein (position 9 of the paired domain). The location of the mutated residue, its conservation in all other paired domain-containing proteins described to date, and the nonconservative nature of the substitution suggest that this mutation is responsible for the phenotype observed in the Spd mouse mutant. PMID- 8406488 TI - Localization of a gene (CMT2A) for autosomal dominant Charcot-Marie-Tooth disease type 2 to chromosome 1p and evidence of genetic heterogeneity. AB - Charcot-Marie-Tooth (CMT) disease type 2 (CMT2) is an inherited peripheral neuropathy characterized by variable age of onset and normal or slightly diminished nerve conduction velocity. CMT2 is pathologically and genetically distinct from CMT type 1 (CMT1). While CMT1 has been shown to be genetically heterogeneous, no chromosomal localization has been established for CMT2. We have performed pedigree linkage analysis in six large autosomal dominant CMT2 families and have demonstrated linkage and heterogeneity to a series of microsatellites (D1S160, D1S170, D1S244, D1S228 and D1S199) in the distal region of the short arm of chromosome 1. Significant evidence for heterogeneity was found using admixture analysis and the two-point lod scores. Admixture analyses using the multipoint results for the markers D1S244, D1S228, and D1S199 supported the two-point findings. Three families, DUK662, DUK1241, and 1523 gave posterior probabilities of 1.0, 0.98, and 0.88 of being of the linked type. Multipoint analysis examining the "linked" families showed that the most favored location for the CMT2A gene is within the interval flanked by D1S244 and D1S228 (odds approximately 70:1 of lying within versus outside that interval). These findings suggest that the CMT2 phenotype is secondary to at least two different genes and demonstrate further heterogeneity in the CMT phenotype. PMID- 8406489 TI - The structure of the human dihydrolipoamide dehydrogenase gene (DLD) and its upstream elements. AB - The structural organization of the gene for the E3 subunit of the human alpha ketoacid dehydrogenase complexes, dihydrolipoamide dehydrogenase (DLD), and its upstream elements have been determined by restriction endonuclease mapping and DNA sequence analysis of overlapping genomic clones. The gene is approximately 20 kb long. It contains 14 exons ranging in size from 69 to 780 bp and 13 introns ranging in size from 93 bp to 7.0 kb. All splice donor and acceptor sites conform to the GT/AG rule. The 5' ends of mRNA transcripts upstream from the translation initiation codon were determined by primer extension assay. A "CAAT box"-like sequence is present at 39 bp upstream of the presumptive cap site and the 5' flanking region has been sequenced up to 2.0 kb upstream. There are several sequences compatible with presumptive promoter elements, including an Sp1 binding site, a nuclear respiratory factor 1 site, two cyclic AMP response element binding sites, and a possible negative response element present in the insulin promoter. A 313-bp segment from -2076 to -1763 is 89% homologous to a recently described pTR5 repetitive element found in the human genome. PMID- 8406490 TI - High-resolution mapping of D16led-1, Gart, Gas-4, Cbr, Pcp-4, and Erg on distal mouse chromosome 16. AB - More than 500 backcross progeny from four intersubspecific backcrosses were typed for six markers on distal mouse chromosome 16. Five of these represented genes that mapped within the Sod-1 to Ets-2 interval, which was shown previously to contain the weaver (wv) gene. The map order, including previously mapped reference markers, is (cen)-D16H21S16-D16Led-1-App-Sod-1-Gart-Gas-4-Cbr++ +-wv Pcp-4-Erg-Ets-2. This gene order recapitulates the order of the genes on human chromosome 21 where known. Two of these markers further define the region containing the weaver gene to a 3.9-cM segment between Cbr and Pcp-4. In addition, Pcp-4 was localized to human chromosome 21 by the presence of a human specific restriction fragment in WAV-17, a mouse-human somatic cell hybrid with human chromosome 21 as the only human contribution. PMID- 8406491 TI - Assignment of the human FAU gene to a subregion of chromosome 11q13. AB - The FAU gene is the cellular homologue of the fox sequence in the Finkel-Biskis Reilly murine sarcoma virus (FBR-MuSV). FAU (for FBR-MuSV associated ubiquitously expressed gene) encodes the ribosomal protein S30 fused to a ubiquitin-like protein. A cosmid clone containing the human FAU gene was used for fluorescence in situ hybridization to metaphase chromosomes. The obtained localization to 11q13 was confirmed by hybridization against a panel of somatic cell hybrids containing different parts of chromosome 11 on a hamster background. FAU was then further mapped, both on a panel of radiation-reduced somatic cell hybrids designed to carry different parts of the 11q13 region and by pulsed-field gel electrophoresis. This fine mapping assigned FAU close to the skeletal muscle glycogen phosphorylase gene (PYGM), in a region that contains several oncogenes as well as the putative tumor suppressor genes MEN1 and ST3. PMID- 8406492 TI - Molecular cytogenetic characterization of the DiGeorge syndrome region using fluorescence in situ hybridization. AB - DiGeorge syndrome (DGS) is a developmental defect characterized by cardiac defects, facial dysmorphism, and mental retardation. Several studies have described a critical region for DGS at 22q11, within which the majority of DGS patients have deletions. We have isolated nine cosmid and three YAC clones using previously described and newly isolated probes that have been shown to be deleted in many DGS patients. Using fluorescence in situ hybridization and digital imaging, we have mapped and ordered these clones relative to the breakpoints of two balanced translocations at 22q11 (one in a DGS patient and one in the unaffected parent of a DGS child). Our data indicate that the breakpoint in the unaffected individual distally limits the DGS critical region (defined as the smallest region of overlap), while proximally the region is limited by repeat rich DNA. The critical region includes the balanced translocation breakpoint of the DGS patient that presumably disrupts the gene causing this syndrome. PMID- 8406493 TI - The human bone sialoprotein gene (IBSP): genomic localization and characterization. AB - We have isolated and partially sequenced the human bone sialoprotein gene (IBSP). IBSP has been sublocalized by in situ hybridization to chromosome 4q28-q31 and is composed of six small exons (51 to 159 bp) and 1 large exon (approximately 2.6 kb). The intron/exon junctions defined by sequence analysis are of class O, retaining an intact coding triplet. Sequence analysis of the 5' upstream region revealed a TATAA (nucleotides -30 to -25 from the transcriptional start point) and a CCAAT (nucleotides -56 to -52) box, both in the reverse orientation. Intron 1 contains interesting structural elements composed of polypyrimidine repeats followed by a poly(AC)n tract. Both types of structural elements have been detected in promoter regions of other genes and have been implicated in transcriptional regulation. Several differences between the previously published cDNA sequence (L. W. Fisher et al., 1990, J. Biol. Chem. 265, 2347-2351) and our sequence have been identified, most of which are contained within the untranslated exon 1. Three base revisions in the coding region include a G to T (Gly to Val, amino acid 195), T to C (Val to Ala, amino acid 268), and T to A (Glu to Asp, amino acid 270). In conclusion, the genomic organization and potential regulatory elements of human IBSP have been elucidated. PMID- 8406494 TI - Allelic variation of reporter gene activation by the HRAS1 minisatellite. AB - We have recently shown that constitutively expressed members of the rel/NF-kappa B family of transcription factors bind the HRAS1 minisatellite, VTRHRAS1. We now report that, like other NF-kappa B binding sites, VTRHRAS1 displays pleiotropic transcriptional regulatory activity that is promoter- and cell-type-specific. Both enhancement and suppression are restricted to the human bladder carcinoma cell line EJ, in which we have previously defined a unique form of NF-kappa B p50. We also observe allelic variation in functional activity: the rare a2.1 allele, one member of a class of HRAS1 alleles overrepresented in the genomes of cancer patients, possesses twofold greater enhancer activity than the low-risk alleles, a0.1, a1, and a2. Finally, VTRHRAS1 enhancer activity is upregulated by the adenovirus E1A 13S gene product, demonstrating the potential of the minisatellite for influencing gene expression through several distinct interactions with the transcriptional apparatus. PMID- 8406495 TI - Characterization of the gene encoding human cone transducin alpha-subunit (GNAT2). AB - The human cone transducin alpha-subunit (GNAT2) gene has been completely characterized. The human GNAT2 transcription unit is 9967-bp in length and consists of eight exons with seven introns. The eight exons are identical to the reported cDNA sequence (Lerea et al., 1989). Northern blot analysis of RNA from human retinas and a retinoblastoma cell line, WERI-RB1, reveals a 1.7-kb transcript for GNAT2. Multiple transcription initiation sites were mapped for human retina and WERI-RB1 RNA by primer extension and S1 nuclease protection assays. This gene has seven initiation sites spanning 31 bp. The sequence upstream of the GNAT2 gene shows a TATA box consensus sequence at -29, a CCAAT box consensus sequence at -58 (reverse orientation), and a sequence (CCATAT) similar to the CCAAT box consensus at -76. The GNAT2 upstream sequence shows no significant identity with the upstream region of the human rod transducin alpha subunit gene (GNAT1) or with the upstream regions of the color visual pigment genes, indicating that the expression of GNAT2 may be regulated differently than these other rod- and cone-specific proteins. PMID- 8406496 TI - Isochores and CpG islands in YAC contigs in human Xq26.1-qter. AB - GC levels were assessed at 37 loci across 30 Mb of Xq26.1-qter, a region physically mapped in overlapping yeast artificial chromosome clones. In 8 Mb of R band Xq26, GC is relatively high (up to 44%) in the proximal 4 Mb and relatively low (40-41%) in the distal 4 Mb. Consistently low GC values (38-41%) are observed in G band Xq27. In contrast, further toward the telomere in Xq28, the GC level rises progressively to reach 52% at 2 to 4 Mb from the end of the chromosome; this region is delimited by low GC loci. Across these regions of Xq, the content of rare-cutter restriction enzyme sites containing CpG, including "CpG islands" in the most completely mapped Xq26-27.1 region, is correlated with GC level. Isochore mapping can thus provide one index of putative gene content across mapped regions. PMID- 8406497 TI - Four novel FBN1 mutations: significance for mutant transcript level and EGF-like domain calcium binding in the pathogenesis of Marfan syndrome. AB - Defects of fibrillin (FBN1), a glycoprotein component of the extracellular microfibril, cause Marfan syndrome. This disorder is characterized by marked inter- and intrafamilial variation in phenotypic severity. To understand the molecular basis for this clinical observation, we have screened the fibrillin gene (FBN1) on chromosome 15, including the newly cloned 5' coding sequence, for disease-producing alterations in a panel of patients with a wide range of manifestations and clinical severity. All the missense mutations identified to date, including two novel mutations discussed here, are associated with classic and moderate to severe disease and occur at residues with putative significance for calcium binding to epidermal growth factor (EGF)-like domains. In contrast, two new mutations that create premature signals for termination of translation of mRNA and are associated with reduction in the amount of mutant allele transcript produce a range of phenotypic severity. The patient with the lowest amount of mutant transcript has the mildest disease. These data support a role for altered calcium binding to EGF-like domains in the pathogenesis of Marfan syndrome and suggest a dominant negative mechanism for the pathogenesis of this disorder. PMID- 8406498 TI - Identification of four novel mutations in the COL4A5 gene of patients with Alport syndrome. AB - The type IV collagen alpha 5 chain (COL4A5) genes of patients with Alport syndrome were tested for major gene rearrangements by Southern blot analysis, using COL4A5 cDNA clones as probes. In addition, individual exons were screened for small mutations by single-strand conformation polymorphism (SSCP) analysis. Four new COL4A5 mutations were detected. A duplication of the nine most 3' located nucleotides of exon 49 and the first nucleotide of intron 49 was identified in the COL4A5 gene of one patient. Two patients displayed single base substitutions leading to, respectively, a proline to threonine and an arginine to glutamine substitution in the C-terminal end. Both substitutions involve amino acids conserved through evolution. In COL4A5 intron 41 a mutation changing the splice acceptor site from AG to AA was identified. All mutations cosegregate with the clinical phenotype of Alport syndrome in affected family members. In a control population of 50 individuals tested by PCR-SSCP these mutations were never identified. Together with two mutations reported previously, a total of six mutations were found in 26 patients with Alport syndrome (23%) after systematic screening of about 30% of the COL4A5 coding region. The clinical features of these six patients are described in detail. PMID- 8406499 TI - A variant family of mouse minor satellite located on the centromeric region of chromosome 2. AB - A variant sequence of minor satellite that exhibits a polymorphism in strains and a subspecies of the mouse by pulsed-field gel analysis has been cloned. A segregation study using intrasubspecific backcross mice reveals that the variant maps to the centromeric region of chromosome 2. A long-range physical analysis of the region shows that the variant family is probably located between the proximal telomere and the major satellite. The sequence probe provides an important marker of the chromosome 2-specific centromere. PMID- 8406500 TI - Localization of the human tripeptidyl peptidase II gene (TPP2) to 13q32-q33 by nonradioactive in situ hybridization and somatic cell hybrids. AB - We have assigned the human tripeptidyl peptidase II (TPP2) gene to chromosome region 13q32-q33 using two different methods. First, a full-length TPP2 cDNA was used as a probe on Southern blots of DNA from a panel of human/rodent somatic cell hybrids. The TPP2 sequences were found to segregate with the human chromosome 13. Second, fluorescence in situ hybridization analysis was performed with the same probe. This analysis supported the chromosome 13 localization and further refined it to region 13q32-q33. PMID- 8406501 TI - Actin-binding protein (ABP-280) filamin gene (FLN) maps telomeric to the color vision locus (R/GCP) and centromeric to G6PD in Xq28. AB - Actin-binding protein-280 (ABP-280) is a dimeric actin filament crosslinking protein that promotes orthogonal branching of actin filaments and links actin filaments to membrane glycoproteins. We have mapped the ABP-280 filamin gene (FLN) to Xq28 by Southern blot analysis of somatic cell hybrid lines, by fluorescence in situ hybridization, and through identification of portions of the FLN gene within cosmids and YACs mapped to Xq28. The FLN gene is found within a 200-kb region centromeric to the G6PD locus and telomeric to DSX52 and the color vision locus. PMID- 8406502 TI - Sequence of the WT1 upstream region including the Wit-1 gene. AB - The Wilms tumor gene WT1 encodes a Cys2His2-type zinc finger protein that can bind DNA and function as a transcriptional regulator. The pathological spectrum of tumorigenesis and various developmental defects produced by different WT1 alterations suggests that WT1 controls a number of subsequent effector genes. To define the role of WT1 in these developmental processes it will be important to elucidate mechanisms that govern expression of WT1 itself. To facilitate mapping of the WT1 promoter region and 5' control elements we have determined the sequence upstream of the WT1 transcription unit. This includes the Wit-1 gene that is transcribed in the opposite direction. PMID- 8406503 TI - Pulsed-field map of Xq13 in the region of the human X inactivation center. AB - We have used human/mouse hybrid cell lines to derive a pulsed-field map of the Xq13 region of the human X chromosome, in the vicinity of the X inactivation center (XIC). We have mapped nine loci within two separate clusters (I and II). Cluster I contains three loci (DXS227, XIST, and DXS128) linked within 1700 kb. This cluster also includes the breakpoint of a translocated X;14 chromosome used to define the proximal border of the XIC region. Cluster II covers an additional 1800 kb and physically links six loci (DXS56, DXS171, DXS325, DXS347, DXS356, and DXS441) located between the XIC and the genes for Menkes disease (MNK) and PGK1. Maps of cluster I loci derived from active (Xa) or inactive (Xi) X chromosomes differed, presumably due to methylation differences between the Xa and Xi. This map provides a basis for examining the organization of the Xq13.2-q13.3 region, in and around the XIC, and will assist in the further cloning of expressed sequences from this region. PMID- 8406504 TI - Chromosome mapping of five human cardiac and skeletal muscle sarcoplasmic reticulum protein genes. AB - Fluorescence in situ hybridization (FISH) experiments were performed using genomic and complementary DNA probes in order to determine the location on human chromosomes for five genes expressed in cardiac and skeletal muscle sarcoplasmic reticulum. The chromosome location of each gene was determined in terms of both cytogenetic bands and fractional chromosome length. The ATP2A2 gene, expressing the SERCA2 isoform of the Ca2+ pump, maps to bands 12q23-q24.1, the phospholamban gene (PLN) to 6q22.1, the human skeletal muscle calsequestrin gene (CASQ1) to band 1q21, the cardiac calsequestrin gene (CASQ2) to bands 1p11-p13.3, and the cardiac calcium release channel gene (RYR2) to the interval between band 1q42.1 (distal) and band 1q43 (proximal). PMID- 8406505 TI - Isolation and characterization of an ornithine aminotransferase-related sequence (OATL3) mapping to 10q26. AB - We used a near full-length human ornithine delta-amino-transferase cDNA, huOAT6, as a probe under low stringency hybridization conditions to identify a new autosomal ornithine delta-aminotransferase-related sequence (OATL3). Cloning and characterization of this sequence reveal it to be a partial nonprocessed pseudogene corresponding to exon 3 and flanking intronic sequences of the ornithine delta-aminotransferase structural gene. Using somatic cell hybrids and fluorescence in situ hybridization, we mapped OATL3 to 10q26, adjacent to the ornithine delta-aminotransferase structural gene locus. PMID- 8406506 TI - Midkine gene (MDK), a gene for prenatal differentiation and neuroregulation, maps to band 11p11.2 by fluorescence in situ hybridization. AB - Midkine (MDK) is a retinoic acid-responsive gene concerned with prenatal development and neurite growth. We mapped the gene to band p11.2 of chromosome 11 through fluorescence in situ hybridization analysis and using a 4.5-kb fragment of its human genomic DNA. PMID- 8406507 TI - Characterization of the chicken and quail homologues of the human gene responsible for the X-linked Kallmann syndrome. AB - The human KAL gene, responsible for the X-linked Kallmann syndrome, was isolated previously. Southern blot analysis using human cDNA probes detected cross hybridization with DNA from several organisms, including chicken and quail. The entire coding sequences of chicken and quail KAL cDNAs were determined. A comparison of these cDNAs with the human KAL cDNA reveals an overall identity of 73 and 72%, respectively. This results in 76 and 75% identity at the protein level. The highest conservation was found in the WAP four-disulfide core motif and in two of the four fibronectin type III repeats reported in the human protein. These results further support the hypothesis that the KAL protein is an extracellular matrix component with anti-protease and adhesion functions. PMID- 8406508 TI - Chromosomal localization and genomic cloning of the mouse alpha-tropomyosin gene Tpm-1. AB - In vertebrates, the alpha-tropomyosin gene, Tpm-1, codes for at least 9 tropomyosin isoforms that are expressed by tissue-specific alternative splicing. Using interspecies backcrosses, we have localized Tpm-1 on mouse chromosome 9, cen-Cyp1a2-Tpm-1-Mod-1-Mylc-Scn5a, near the d-se region. The restriction fragment length variant used for chromosomal assignment, as well as other restriction fragments hybridizing to a 3'-specific alpha-tropomyosin cDNA probe in genomic Southern blots, was investigated by cloning 17.5-kb of Tpm-1. The same restriction patterns were observed, proving the identity of the mapped and the cloned gene. The identity was supported by sequencing the 3' end of the gene. PMID- 8406509 TI - Localization of a second NM23 gene, NME2, to chromosome 17q21-q22. AB - NM23 is a candidate tumor suppressor protein and has recently been identified as an NDP kinase. The expression of NM23 is inversely related to the metastatic potential of tumor cells. Two NM23 genes, NME1 and NME2, that code for the A and B chains of the kinase, respectively, have been cloned. To determine the human chromosomal location of the NME2 gene, we have analyzed DNA from rodent-human cell lines and hybrid cell lines containing portions of chromosome 17 by a combination of PCR amplification and Southern hybridization. The NME2 gene was mapped to the chromosome region 17q21-q22, the same region in which the NME1 gene has been localized. This region is linked to the early onset breast/ovarian locus (BRCA1) and allelic deletions of NME1 have been associated with metastatic potential of colorectal carcinomas. PMID- 8406510 TI - Galactosemia caused by a point mutation that activates cryptic donor splice site in the galactose-1-phosphate uridyltransferase gene. PMID- 8406511 TI - Physical and genetic localization of the gamma subunit of the cyclic GMP phosphodiesterase on the long arm of chromosome 17 (17q25). PMID- 8406512 TI - Assignment of the human deoxycytidine kinase (DCK) gene to chromosome 4 band q13.3-q21.1. PMID- 8406513 TI - Strategies in cDNA programs. PMID- 8406514 TI - Positioning of the autochthonous Aran Valley population among Basque and Pyrenean people by means of ABO, Rh (D) and Duffy blood group determinations. AB - A total of 205 unrelated autochthonous individuals of both sexes from the population of the Aran Valley in the central Pyrenees were tested for ABO, Rh and duffy blood groups. This population is of interest because of its relative geographical and historical isolation and its specific peculiarities, such as its own language. The results show a good correlation between the principal component analysis graphic and the geographic positions of the Basque and Pyrenean populations with which the Aranese population is compared. PMID- 8406515 TI - Orosomucoid polymorphism in Finns, Swedes and Swedish Saamis. AB - Genetic polymorphism of orosomucoid (ORM) was studied by isoelectric focusing and immunoblotting in Finns, Swedes and Swedish Saamis. The ORM2 locus was found to be monomorphic in all three ethnic groups. In the Swedish sample the frequency of the ORM1*2 allele (0.414) was within the range found in other European populations, whereas Finns (0.282) and Saamis (0.210) showed significantly lower ORM1*2 frequencies. The extremely low ORM1*2 frequency in the Saamis further underlines the genetic uniqueness of this population. The ORM1*2 frequency in Saamis resembles those in Asiatic Mongoloid populations, but this is unlikely to reflect an Asiatic influence, since the accumulated knowledge on genetic markers in the Saamis show no unequivocal evidence for an Asiatic influence in this population. PMID- 8406516 TI - Power of affected sibling method tests for linkage. AB - When the mode of inheritance of a disease susceptibility (DS) gene is unknown, the affected sibling method can be applied to study whether a DS gene and a marker gene are linked. This method considers how k affected siblings (k > or = 2) share marker alleles identically by descent from their parents. Several nonparametric and likelihood ratio test statistics have been proposed for the use with affected sibling marker gene data. We compare the power of these proposed tests for linkage under different genetic models via simulation. We find that, in general, the likelihood ratio tests are slightly more powerful, but the gain in power may not warrant the additional computation burden imposed. PMID- 8406517 TI - Familial amyloidotic polyneuropathy in Sweden: geographical distribution, age of onset, and prevalence. AB - Familial amyloidotic polyneuropathy (FAP) in Swedish patients is associated with the same transthyretin mutation (TTRMet30) as in Portuguese, Japanese, Brazilian and Majorcan patients. Yet, the age of onset of FAP is much later in Sweden than in other populations. We have studied 239 cases of FAP from northern Sweden, their geographical distribution, differences in age of onset, and estimated prevalence and incidence rates. Cases and families concentrate mainly in two areas, around the towns of Skelleftea and Pitea. Mean age of onset was found to be later in the Pitea (58.8 +/- 10.8) than in the Skelleftea area (54.4 +/- 13.5). Unusually high figures were found for prevalence rates (91 x 10(-5) and 104 x 10(-5), respectively) in 1985. Mean yearly incidences were 3.1 x 10(-5) and 4.4 x 10(-5), respectively, over the period 1985-1989. PMID- 8406518 TI - Identification of three novel cystic fibrosis mutations in a sample of Italian cystic fibrosis patients. AB - An analysis of 274 non-delta F508 Italian cystic fibrosis chromosomes was carried out to determine their molecular defect. In a first step, the delta F508 and 59 other mutations were detected by polyacrylamide gel electrophoresis, restriction digestion, and the amplification refractory mutation system (ARMS) technique. The molecular defects of the other chromosomes were screened for by denaturing gradient gel electrophoresis analysis of exons 3, 4, 7, 11, 12, 13, 14a, 17b, 19 and 20. Direct sequencing was carried out if necessary. This approach allowed us to identify 3 novel mutations, namely M348K, D614G and F693L. PMID- 8406519 TI - ABO, Rh, MNSs, sex and typhoid fever. AB - A case-control study on the association between genetic markers (ABO, Rh, MNSs and sex) and typhoid fever (TF) was performed. We found that (a) the B allele conferred protection to females for TF and a mild susceptibility to males; (b) the CDe haplotype (or the RH3 phenotype, mostly CDe/CDe) was associated with protection against Salmonella in both sexes, while cDE (or RH7, mostly cDE/cDE, and RH8, mostly cDE/cde) was associated with susceptibility to TF, and (c) heterozygotes Ss (MNSs system) had a high susceptibility, while SS and particularly ss homozygotes seemed to be protected for the disease. There were very significant interactions among the blood systems and sex. PMID- 8406520 TI - Study of HLA antigens in patients with osteosarcoma. AB - HLA typing was carried out in a group of twenty-four Spanish unrelated patients diagnosed with conventional high-grade osteosarcoma. All of them were typed for HLA-A and HLA-B antigens. A significant increase in the HLA-A11 (33.3 vs. 12%, chi 2 = 9.752, corrected p value = 0.016) and HLA-B7 (33.3 vs. 11.3%, chi 2 = 10.919, corrected p value = 0.011) was found in osteosarcoma patients compared with the control group. A trend towards an increased frequency in HLA-A24 and HLA A28 antigens was also found. PMID- 8406521 TI - Human dopamine transporter gene not linked to schizophrenia in multigenerational pedigrees. AB - A large body of data suggests that perturbations in brain dopaminergic transmission play a role in the pathogenesis of schizophrenia. Recently, the gene for the human dopamine transporter has been cloned and polymorphisms have been identified. Because mutations of the dopamine transporter gene might underlie the cause of schizophrenia in a subset of families, we undertook a linkage analysis between schizophrenia in 9 families and a dopamine transporter gene polymorphism. Evidence of linkage was not found in most families assuming autosomal dominant or recessive inheritance. PMID- 8406522 TI - Haptoglobin (HP), transferrin (TF) and group-specific component (GC) subtype distribution in Bantu-speaking people from Malawi. AB - A sample from Malawi was studied for the genetic markers haptoglobin (HP), group specific component (GC) and transferrin (TF). The following allele frequencies were found. For HP: 1F = 0.355, 1S = 0.204, 2FS = 0.396, 2SS = 0.044; the allele 2FF was not observed. For GC: 1F = 0.814, 1S = 0.057, 2 = 0.079, 1A1 = 0.047, 2A1 = 0.0025. For TF: C1 = 0.894, C2 = 0.075, C3 = 0.0026, D1 = 0.029. The HP subtype distribution is among the first to be reported for African blacks. PMID- 8406523 TI - Distribution of the HLA-DQ alpha alleles and genotypes in a sample of a population from Barcelona (Spain). AB - Allele and genotype frequencies at the HLA-DQ alpha locus were determined by the use of polymerase chain reaction (PCR) and the reverse dot blot format. 6 HLA-DQ alpha alleles and 21 genotypes were detected among the 178 unrelated individuals from Barcelona. A fair agreement was found between observed and expected numbers assuming a Hardy-Weinberg equilibrium. The Barcelona population is statistically similar to the Finnish population, but differs significantly from the Japanese, Black and Caucasoid series. PMID- 8406524 TI - When your patient is a nurse. AB - While stereotypes exist about patients who are nurses, it is very important that nurses who are patients be treated with respect, concern, and in need of professional nursing care, just as any patient should be. Just because a patient happens to be a nurse does not mean care should be given any differently. PMID- 8406525 TI - Humor ... the best medicine, or is it? PMID- 8406526 TI - A childhood dream realized. PMID- 8406527 TI - Dianna Saenz. Interview by Amanda Sue Niskar. PMID- 8406528 TI - Camp nursing as a clinical elective. PMID- 8406529 TI - "No code"--the meaning behind the words. PMID- 8406530 TI - A beginning, not an end.... PMID- 8406531 TI - Getting out of the fat lane. PMID- 8406532 TI - Overview: the International Council of Nurses: the past, present and future. PMID- 8406533 TI - ANA, a vital connection to international nursing. PMID- 8406534 TI - Cultural immersion: student experiences in Mexico. PMID- 8406535 TI - Reflections of a chemistry tutor. PMID- 8406536 TI - Louise's lesson. PMID- 8406537 TI - American student participation in ICN and international affairs. PMID- 8406538 TI - Nursing education in the United Kingdom. PMID- 8406539 TI - Looking north: a view of Canada's health care system. PMID- 8406540 TI - Retrospective: Fuld Fellows in Vienna. PMID- 8406541 TI - International nursing: a student outlook. AB - World health depends upon our field of vision. If we selectively view the world from a narrow perspective, we will be unable to function effectively as nurses. The sobering reality of health conditions throughout the world should awaken our consciousness and sharpen our focus on priorities for the future. We can agree with Lindquist, that "nursing must be viewed from a global perspective if it is to influence the quality of health care provided in the future." Although not every student has the means and availability to travel overseas in a volunteer capacity, students may begin to examine the possibilities and start their own correspondence with an international agency. Regardless of our realm of service, the opportunity to provide care for individuals of all cultures, whether abroad or at home, remains the highest privilege of our profession. PMID- 8406542 TI - Lipid molecular shapes and membrane architecture. AB - A simple biomembrane like erythrocyte contains well over hundred lipid species with diverse molecular shapes differing in the number of acyl chains, chain length, unsaturation and head group composition. A delicate balance between these molecular shapes is necessary in order to have a functional membrane. It is well established that the activities of a number of membrane-bound enzymes and other properties such as aggregation, spontaneous vesiculation, pathophysiological properties and lipid-protein interactions of lipids depend on the acyl chain length, unsaturation and head group composition. In fact, the molecular shape of a phospholipid molecule, as modulated by changes in chain length, unsaturation and head group composition, is probably what is affecting the above mentioned properties. The molecular shape of a lipid depends on a dimensionless packing parameter, S, the value of which influences the size and shape of aggregate formed upon hydration. In fact, the additivity of S values of lipid mixtures explains a number of experimental observations. The molecular shape concept, although very simple, explains many membrane phenomena like complementarity of molecular shapes of non-bilayer lipids to form stable bilayers. Membrane permeability is controlled to a large extent by lipid packing which depends upon molecular shapes. In fact, membranes maintain their lamellar structure by delicately balancing the composition of bilayer-forming and non-bilayer-forming lipids indicating that complementarity of molecular shapes is essential to maintain the permeability barrier. Based on this, the complementary molecular shape model of cell membrane is proposed. PMID- 8406543 TI - Isolation of a salt-inducible gene from rice and its molecular cloning. AB - The genomic sequences of a salt inducible gene of rice have been isolated from both Indica and Japonica varieties of rice. The cDNA of this gene, SalT, characterized earlier shows sequence identity with both the genomic sequences. Approximately 750 base pairs of intervening sequence is present in the 1.2kb genomic sequence, obtained by PCR. There is a high degree of homology between the Indica and Japonica SalT genes except for the presence of an extra HindIII site in the former. A genomic library of Indica rice was screened and a phage clone of the SalT gene was partially characterized. Different species of monocot and dicot plant DNA showed sequence homology with SalT gene. PMID- 8406544 TI - Molecular cloning and restriction endonuclease analysis of 0.4 kb Hin dIII'O' fragment of bovine herpesvirus 1 DNA. AB - Bovine herpesvirus 1 DNA has been isolated by SDS lysis of the virus purified from potassium tartrate (10-50%) density gradient centrifugation. The quality and quantity of viral DNA was checked by UV spectrophotometry and ethidium bromide stained agarose gel electrophoresis. The 0.4 kb Hin dIII'O' fragment of BHV-1 DNA was selectively cloned into Hin dIII cut pUC9 plasmid DNA (2.665 kb). Recombinants were screened by white/blue colonies as well as Hin dIII restriction enzyme analysis. On restriction endonuclease analysis of recombinant plasmid DNA (p-BH-0) with several restriction enzymes, viz., Sau 3A, Hin fI, Rsa I, Sal I, Dra I, Bgl I, Bgl II, Sma I, Hpa I, Stu I, Mlu I, Xho I, Kpn I, Hae III, Eco RI, Bam HI, Pst I, Pal I, revealed insert viral DNA having sites for Hin fI, Hae III, Rsa I, Sma I, only. Further, the partial restriction map of the recombinant plasmid DNA was constructed using above enzymes. PMID- 8406545 TI - An invertase with unusual properties secreted by sucrose-grown cells of Corynebacterium murisepticum. AB - The mode of sucrose utilisation by Corynebacterium murisepticum cells growing on M9 minimal medium supplemented with 0.4% sucrose as the carbon source was studied. It was observed that during growth of this organism, sucrose in the medium is hydrolysed to glucose and fructose, suggesting the formation of an extracellular invertase. Unlike in other microorganisms (e.g. Saccharomyces cerevisiae) the invertase formation is not repressed by the presence of glucose in the medium. The invertase was found to be the only predominant extracellular protein in the culture broth and could be purified in a single step by precipitation at 90% ammonium sulphate saturation. The purified protein had a molecular mass of 70,000 daltons. It not only showed invertase activity, but also a fructosyltransferase activity as it could convert sucrose to beta-1,2 difructose, as well as to glucose and fructose. PMID- 8406546 TI - Liposomes as a carrier for mannophosphoinositide antigens of mycobacteria. AB - Phosphatidylcholine liposomes have been used as carriers of mannophosphoinositides (PIMs) of mycobacteria to examine their immunological properties. PIMs incorporated in egg phosphatidylcholine (EPC) liposomes elicited both humoral and cell-mediated immune responses in mice. Addition of cholesterol at 43 mole% to PC enhanced the immune responses while the reverse was observed with EPC liposomes bearing negative or positive charge. Liposomes made of dioleoylphosphatidylcholine (DOPC) with cholesterol (43 mole%) proved to be a better immunoadjuvant with this antigen as compared to those made of EPC or dipalmitoylphosphatidylcholine (DPPC). Our results suggest that DOPC liposomes containing cholesterol are better carriers for PIMs-antigen as compared to Freund's incomplete adjuvant. PMID- 8406547 TI - Human plasma low density lipoprotein: a fluorescence study. AB - Hydrophobic nature of human plasma low density lipoprotein surface has been studied by fluorescence spectroscopic method. Enhancement in 8-anilino-1 naphthalene sulphonate (ANS) fluorescence quantum yield from 0.004 to 0.114 at 470 nm has suggested that the ANS binding sites are fairly low in polarity. LDL has been found to have 77 homogeneous binding sites for ANS (Ka = 2.5 x 10(5) M 1). The binding of an ANS molecule does not affect the successive binding sites. Variation in temperature from 15 degrees to 45 degrees C did neither alter the number of binding sites nor association constant. Quenching of protein fluorescence (lambda exc 286 nm, lambda ems 336 nm) indicated the occurrence of energy transfer in LDL-ANS complex arising from conformational changes capable of bringing charge acceptor segments near to other ANS site. About 30-fold increase in ANS quantum yield and large shift in the emission maximum are characteristic features of large hydrophobic environment on the surface of LDL particle. PMID- 8406548 TI - Modulation of monkey small intestinal brush border membrane D-glucose transport by nonesterified fatty acids. AB - Brush border membranes isolated from monkey intestinal mucosa was found to contain considerable amount of nonesterified fatty acids. Incubation of brush border membranes with fatty acid free albumin selectively removed the free fatty acids more than 80% without altering the level of phospholipids or cholesterol. The sodium dependent D-glucose transport was stimulated by the albumin treatment. Kinetic study showed that albumin treatment did not alter the apparent affinity (Km) of the transporter for glucose whereas the maximal velocity (Vmax) was increased significantly. The sodium dependent D-glucose transport was inhibited by the exogenously added unsaturated fatty acids. Saturated fatty acids and methyl esters of unsaturated fatty acids showed no inhibition. Based on these results, it may be concluded that free fatty acids inhibit the sodium dependent intestinal D-glucose transport either by directly interacting with the transport protein or by abolishing the sodium gradient. PMID- 8406549 TI - Alterations in rat intestinal sucrase and alkaline phosphatase activities in alloxan induced experimental diabetes. AB - Rat intestines revealed a significant loss of proteins after seven days of alloxan induced diabetes. The data suggested the presence of two forms of alkaline phosphatase (ALP) in normal rat intestines. Along with the loss of proteins from the intestines during diabetes, a form of ALP which appears to be loosely bound to the intestine is also flushed out. Total brush border membrane (BBM) proteins are relatively preserved from such leaching effect of alloxan induced diabetes. Thus, sucrase and another form of ALP which appears to be strongly bound to the BBM are flushed out at a slower rate as compared to the other intestinal proteins and loosely bound soluble ALP. BBM preparations from diabetic rat intestines showed lower ratios for BBM/intestinal homogenate sucrase or ALP activity/mg proteins as compared to the normal control rats. Such ratios, therefore, misdepict the purity as low for the BBM from diabetic rats which is merely because of the decreased contents of proteins in the intestinal homogenate during alloxan-induced acute experimental diabetes. PMID- 8406550 TI - Quantitative structure-activity relationship studies on benzodiazepine receptor binding: investigation of interaction model. AB - With a view to providing perfection to the benzodiazepine receptor model proposed earlier [S P Gupta, R N Saha & V Mulchandani (1992) J. Mol. Recog, 5, 75-80] a few more QSAR studies on a series of 9-benzylpurines and tetracyclic 1,4 benzodiazepine derivatives have been made. The models showing the interaction of these compounds with the receptor are proposed. It is found that the receptor model, unlike the one proposed earlier, requires the presence of a polar site along with all the other essential sites. PMID- 8406551 TI - Effect of some pesticides/weedicides on cathepsin B activity and lysosomal membrane. AB - The in vitro inhibitory effects of various weedicides and pesticides on goat brain cathepsin B and their labilizing potency on the lysosomal membrane were quantitated. Endosulfan an organochlorine insecticide inhibited the enzymic activity to approximately 50% at 7 mM concentration followed by methyl parathion, aldrin, melathion and benzene hexachloride (BHC) in that order. Among the weedicides, butachlor was found to be most inhibitory (approximately 50% activity was lost at 6 mM) followed by isoproturone (28%) and anilophos (19%). When the labilizing/stabilizing potency of all these drugs was observed on lysosomal membrane it was found that none of these was capable of stabilizing the membrane. At 40 degrees C and 1 mM drug concentration, aldrin, endosulfan, melathion and anilophos were found to be strong labilizers of the lysosomal membrane. Others like isoproturone, BHC and methyl parathion had moderate labilizing effect. The labilization potency of the drugs was temperature dependent and was less pronounced at 25 degrees C as compared to 40 degrees C. PMID- 8406552 TI - Cross-binding activity and protective capacity of monoclonal antibodies to lipid A. AB - Six hybridoma clones (1M, 4M, 9M, 11M, 18M and 31G), secreting monoclonal antibodies (mAbs) against lipid A were obtained after fusion between cells of mouse myeloma line X63-Ag8.653 and spleen cells from BALB/c mice immunized with acid treated Salmonella minnesota bacteria coated with additional free lipid A. The specificity and cross-binding activity of the mAbs were characterized in ELISA by using synthetic lipid A analogs as well as different lipid A and lipopolysaccharides (LPS) extracted from R- and S-form bacteria. It was found that the antibodies recognize epitopes in which phosphate groups, especially those at the C4' position of the glucosamine backbone of lipid A, were present. These epitopes were accessible also for the antibodies in purified intact LPS. By using a set of core glycolipids with increasing completion of the core region of the molecule and S-LPSs it was shown that the mAbs cross-reacted with a variety of R- and S-form LPS. The binding activity decreased with increasing length of the polysaccharide chain. The mAb did not prevent ultimate lethality of mice challenged with Klebsiella pneumoniae B and Salmonella typhimurium C5. However a delay of mortality rate of mice pretreated with antibodies 18M and 31G and infected with K. pneumoniae was seen. PMID- 8406553 TI - An anti-digoxin monoclonal antibody seems to express more than one functional paratope. AB - An anti-digoxin monoclonal antibody (mAb 4G3) has been produced and characterized with respect to its fine specificity and affinity. In an independent series of experiments anti-idiotypic monoclonal antibody (mAb 7G9) was selected which reacted with the antigen-binding center of an anti-human chorionic gonadotropin monoclonal antibody (anti-hCG mAb 1B10). In detailed studies on its binding characteristics it has been shown that mAb 4G3 binds to an anti-idiotypic monoclonal antibody mAb 7G9 in solution. Western blotting experiments showed that mAb 4G3 reacted against antiidiotypic antibody under non-reducing conditions, only. Moreover, mAb 4G3 has been shown to express self-binding properties. Absorption with saturating amounts of its specific hapten, i.e. digoxin, did not change the binding of mAb 4G3 to anti-idiotypic antibody and its self-binding ability. It is speculated on the basis of these data that mAb 4G3 possesses more than one functional paratope. PMID- 8406554 TI - Spontaneous apoptosis in NS-1 myeloma cultures: effects of cell density, conditioned medium and acid pH. AB - Apoptosis is a form of cell death which plays an important role in many biological processes including the regulation of B and T lymphocyte functions. We report here the spontaneous development of extensive apoptosis in cultures of the NS-1 mouse myeloma cell line following overgrowth. The apoptosis was identified by both its ultrastructural features and its DNA fragmentation pattern. High cell density and conditioned medium, but not acid pH, were found to be major inducers of apoptosis in this experimental system. PMID- 8406555 TI - In vivo distribution of particulate antigens and liposomes in murine spleen. A possible role in the humoral immune response. AB - Several particulate antigens and liposomes were intravenously injected in mice in order to study their localization patterns in spleen and liver. Liposomes have been proposed as promising carriers for haptens and antigens. It was studied whether the phospholipid composition, cholesterol content and charge of the liposomes played a role in their distribution within the spleen. Different thymus independent type 1 and type 2 and thymus-dependent particulate antigens as well as liposomes were labeled with the lipophilic fluorochrome Di-I. After labeling they were intravenously injected and spleens and livers were removed at different time intervals and prepared for light- and fluorescence-microscopy. We have observed that all particulate antigens and liposomes administered to the mice localized according to the same distribution pattern in the spleen. After 2 and 4 h particles were located in macrophages of the marginal zone and after 24 h white pulp macrophages had also ingested particulate antigens and liposomes. So we conclude that the distribution of the particulate antigens and liposomes in the spleen is independent of the immunological nature of the particles. Results are discussed with respect to the question whether or not the distribution of particulate antigens and liposome associated antigens or haptens, may be a crucial factor in determining the type of immune response to be elicited. PMID- 8406556 TI - Masking of HLA class I molecules expressed on K-562 target cells can restore their susceptibility to NK cell cytolysis. AB - The aim of our study was to follow whether class I HLA antigens play a role in the susceptibility of K-562 target cells to NK cell lysis as well as in the interaction between NK and target cells. After K-562 cells were treated with rIFN gamma, HLA class I antigens appeared on their surface. Those HLA class I+, in comparison to HLA class I-, K562 target cells became about 40-50% less susceptible to NK cell cytolysis. This influence of HLA class I antigens was abrogated when class I+ K-562 cells were incubated with anti-HLA class I monoclonal antibodies Bra 23/9. The restoration of susceptibility of these targets to NK cell cytolysis was not caused by ADCC mechanism as determined by F(ab')2 fragments or by mAb CD16. Further, the percentage of conjugates between effector and target cells was not significantly altered neither when HLA class I antigens appeared on the surface of K-562 target cells nor when HLA class I+ K 562 cells were incubated with anti-HLA class I monoclonal antibodies. To study this question, we used a different approach because both expression of HLA class I molecules and their masking with appropriate monoclonal antibody were done by means of the same cell line of target cells. PMID- 8406557 TI - MHC-specific graft-protective and delayed-type hypersensitivity (DTH) suppressive activity of a CD4+ CD8+, alpha beta T cell receptor (TCR) positive lymphoma isolated from a tolerant mouse. AB - Two lymphomas were found in, and isolated from A (H-2a) mice in which permanent transplantation tolerance was induced to CBA (H-2k) histocompatibility antigens by the neonatal injection of (CBAxA)F1 spleen cells. They proved to be of recipient origin and were transferable to syngeneic A mice, growing as disseminated lymphomas (L33 and L46) and killing the recipients rapidly. Analysis of the cell surface antigens disclosed that both lymphomas had an immature T cell phenotype [Thy-1+, CD5+, CD3low, TCR alpha beta low, CD4low, CD8high, heat-stable antigen (HSA) positive, and CD44-, MHC class II-, CD45R-, sIg-, Gr-1-, CD11b-]. Intraperitoneal (i.p.) injection of syngeneic A mice with viable L33 lymphoma cells resulted in a dose-dependent, significant prolongation of the mean survival times of "specific" CBA and MHC-identical B10.BR skin allografts as compared to the survival of appropriate grafts in non-lymphoma-bearing controls. The survival times of third party MHC-incompatible B10 (H-2b) and B10.D2 (H-2d) allografts were only slightly prolonged in A mice inoculated with L33 cells. The graft protective effect was not abrogated if the proliferative capacity of the L33 cells was blocked by in vitro mitomycin C (MMC) pretreatment. Furthermore, the inoculation of L33 lymphoma into A mice significantly inhibited their DTH response to the sensitizing CBA histocompatibility antigens. In contrast, the L46 lymphoma had no effect on the survival of CBA allografts and the DTH reactivity. These data suggest that the CD4+CD8+TCR alpha beta + L33 T cell lymphoma originating from a neonatally tolerant mouse has a specific immunosuppressive effect on the in vivo reactivity of syngeneic mice to the tolerance-inducing (MHC class I) alloantigens. PMID- 8406558 TI - In vitro effect of recombinant interferon gamma in combination with LPS on amoebicidal activity of murine Kupffer cells. AB - The present study examines the role of liver macrophages (Kupffer cells), of C57BL/6 mice, as effector cells responsible for the killing of Entamoeba histolytica trophozoites in vitro. It was shown that unstimulated Kupffer cells were inefficient in the killing of E. histolytica trophozoites in vitro. Interferon gamma (IFN-gamma) alone was not able to activate Kupffer cells to amoebicidal state. However, Interferon gamma and lipopolysaccharide (LPS) acted synergistically in this phenomenon. It seems that the acquisition of amoebicidal activity is associated with the involvement of hydrogen peroxide, because the addition of catalase partially decreases the killing of this parasite by Kupffer cells. In addition, it appears that the amoebicidal activity of IFN-gamma-treated Kupffer cells is contact-dependent. Our results indicate that the immunologic production of IFN-gamma is important in the activation of Kupffer cells for controlling this parasite and that Kupffer cells are strong effector cells against the amoebae. PMID- 8406559 TI - Induction of mouse IgA B cell differentiation by phorbol ester in the absence of proliferation. AB - Previously, we have demonstrated that protein kinase C-activating phorbol esters selectively induce IgA synthesis by mouse B cells whether stimulated or not by lipopolysaccharide (LPS). In this study, we investigated in detail whether this maturation into IgA-secreting cells is preceded by cell proliferation. T-depleted spleen B cell suspensions were fractionated by Percoll gradient into a dense fraction that contained a majority of small resting B cells, and a hypodense fraction relatively enriched in large B cells. Phorbol 12-myristate, 13-acetate (PMA) induced IgA synthesis in both types of cells, though to a greater extent in large B cells. The phorbol ester accelerated and increased DNA synthesis in LPS stimulated B cells but did not induce any DNA synthesis in small or large B cells, while at the same time decreasing the percentage of cells in the S, G2/M phases of the cell cycle. Inhibition of proliferation by colcemid, a cell cycle blocker, did not prevent PMA-induced IgA synthesis. The number of IgA secreting cells determined by the enzyme-linked immunospot reached a maximum at 24 h, by which time more than 50% of IgA B cells showed morphological features characteristic of plasma cells. No increase in percentage of large or blastic IgA cells was observed. Collectively the data indicate that PMA induces the terminal differentiation of mIgA+ B lymphocytes into IgA plasma cells without any DNA synthesis and cell proliferation. PMID- 8406560 TI - Effect of L-alanine and some other amino acids on thymocyte proliferation in vivo. AB - L-alanine was shown earlier to play a significant role for the proliferation of lymphocytes in vitro. In the present work the effect of L-alanine and some other amino acids on thymocyte proliferation was studied in vivo by local administration into one thymus lobe of guinea pigs. Proliferating cells were pulse labelled with bromodeoxyuridine (BrdUrd). The labelling index of the treated lobe significantly exceeded that of the contralateral, control lobe at 48 h after treatment with 10 or 100 micrograms L-alanine, indicating stimulated proliferation. The higher dose, which was also tested after other time intervals, stimulated also at 24 h. The difference in proliferative activity between the lobes was verified by mitotic studies. The effect of L-alanine was mainly on the large, low density, highly proliferating precursor cells. No other amino acids tested (D-alanine, cysteine, hydroxyproline, serine, tryptophan), or pyruvate produced significant differences between the treated and control lobes. PMID- 8406561 TI - Surface expression of Forssman glycosphingolipid antigen on murine bone marrow derived macrophages is subject to both temporal and population-specific regulation and is modulated by IL-4 and IL-6. AB - The Forssman glycolipid antigen (Fo) has been shown to be a differentiation marker for mouse macrophages both in vivo and in vitro. In order to determine whether or not there is a relationship between stage of differentiation and Fo expression, we have analyzed the kinetics of Fo expression during the growth of cultured mouse bone marrow-derived macrophages (BMDM). BMDM were grown in serum free medium to avoid the possible influence of undefined serum factors. In this medium they could be maintained over a period of up to 20 days with cell yields comparable to those obtained with serum-supplemented media. Fo antigen was assayed with a specific antibody using both a whole cell ELISA and immunocytochemical staining of cells grown on slides. With increasing age in culture, BMDM showed a gradual quantitative increase in Fo expression and parallel increase in the Fo+ BMDM fraction from about 10% Fo+ cells on the 10th day of culture to a maximum of 50%-60% Fo+ cells between the 17th and 19th days. The temporal control over the development of the Fo+ cell fraction was intrinsic to BMDM maturation but was specific for Fo. During the same time period expression of MHC class II (Ia) remained consistently low, whereas expression of both Mac-1 (C3bR) and the macrophage-specific marker ER-BMDM-1 was always high. The interleukins IL-4 and especially IL-6 induced a premature expression of Fo at earlier stages of BMDM culture, but neither could promote further Fo expression once the intrinsically occurring maximum had been reached. No evidence in support of an autocrine regulation of Fo expression by IL-6 could be obtained, nor could a connection between cell cycle status and Fo expression be established. These data provide further evidence that Fo is a temporally regulated differentiation marker for a mouse macrophage subpopulation and for modulation of its expression by lymphokines. PMID- 8406562 TI - Analyses of origin of synovial cells and repairing mechanisms of arthritis by allogeneic bone marrow transplantation. AB - MRL/lpr mice spontaneously develop rheumatoid arthritis (RA)-like disease. Recently we have observed that bone marrow transplantation (BMT) with bone graft to recruit donor stromal cells can be used to treat autoimmune diseases (including RA-like lesions) in MRL/lpr mice. In this paper, we characterize the origin of synovial cells with the use of radiation chimeras and elucidate the repairing mechanism of RA by BMT. Type A synoviocytes have been thought to play an important role in the initiation of inflamed synovia, since a large number of Type A synoviocytes have been seen in inflamed synovia of both RA patients and MRL/lpr mice. Using [C57BL/6JJic-bg-->MRL/lpr] chimeric mice, we found Type A synoviocytes to be derived from donor bone marrow cells. They appeared in the inflamed synovia 4 weeks after BMT. However, at this time, the repairing process was not prominent. Serial biopsy studies revealed that newly developed T cells with normal functions play a more crucial role in the treatment of RA in MRL/lpr mice than do Type A synoviocytes. PMID- 8406563 TI - Identification and quantification of complement regulator CD46 on normal human tissues. AB - CD46 is a cell-surface regulatory molecule that prevents lysis of autologous human cells by activated complement. It has been well characterized on leucocytes, reproductive cells and various cultured cell lines and is considered to be ubiquitously expressed. We now extend these analyses and describe CD46 in a variety of different human tissues. Strong expression was observed by immunohistology on epithelial cells lining exocrine ducts and glands, such as salivary gland and pancreas and on kidney tubules and glomerular epithelium. Quantitative tissue expression was measured by radioimmunoassay and confirmed histological observations. Thus, CD46 is highly expressed on cells in contact with extracellular fluids thought not to contain large quantities of complement but which may still be subjected to complement attack thereby necessitating the presence of complement regulators to prevent non-specific destruction of cells. PMID- 8406564 TI - Complement C3 gene expression and regulation in human glomerular epithelial cells. AB - Extra-hepatic synthesis of complement is thought to mediate local tissue inflammatory injury. To investigate this phenomenon in the glomerular epithelial cell (GEC), we examined the biosynthesis and regulation of gene expression of the third component of complement in isolated human GEC derived from normal tissue. Metabolic labelling and immunoprecipitation studies demonstrated that C3 protein was synthesized, processed and secreted by GEC under basal conditions. The secreted C3 alpha and beta polypeptide chains had identical electrophoretic mobilities with those of hepatic C3. Examination of cellular RNA using semi quantitative polymerase chain reaction (PCR) showed that C3 gene expression was present in unstimulated GEC and was increased by stimulation with interferon gamma (IFN-gamma) in a time- and dose-dependent manner. Tumour necrosis factor alpha (TNF-alpha), while mediating an increase in monocyte U937 C3 expression, revealed no evidence of regulation of GEC C3 gene expression. These results indicate that human GEC spontaneously express the C3 gene and that increased gene expression is regulated by IFN-gamma. These observations may reflect part of a wider mechanism of protection against or mediation of local, immune-mediated tissue injury. PMID- 8406565 TI - Cross-reactive idiotopes among anti-foot and mouth disease virus neutralizing antibodies. AB - Foot and mouth disease virus (FMDV) viral protein 1 is the only one of the four viral proteins (VP) that induces neutralizing antibodies as an isolated protein. A 32 amino acid (AA) residue (32dimer) of FMDV subtype A12 Lp ab VP1 (AA 137-168) was immunogenic against the A12 subtype. Three antibody populations each recognizing different epitopes on 32dimer were isolated by affinity chromatography (AFC) from the serum of a steer which had been immunized with the 32dimer. The 32dimer contains an AA sequence that is recognized by a protective paratope carried on a murine monoclonal antibody (mAb) (7SF-3.H3.1). Polyclonal anti-7SF-3 idiotype antibodies specifically inhibited the binding activity of one of these anti-32dimer antibody populations suggesting the existence of cross reactive paratopic-related idiotopes between mAb 7SF-3 and antibodies elicited by the 32dimer. These anti-idiotypic antibodies were used in AFC to purify antibodies from the anti-32dimer serum. The purified antibody population has characteristics that resemble those of the mAb 7SF-3, i.e. its reactivity with FMDV A subtypes in ELISA, radioimmunoassay (RIA), mouse neutralization and its lack of reactivity with a mAb 7SF-3 neutralizing escape virus variant. Furthermore, these antibodies were specifically inhibited by either anti-mAb 7SF 3 idiotypic antibodies or peptides containing the mAb 7SF-3 epitope. Using the same experimental approach, mAb 7SF-3 idiotope-bearing antibodies were shown to be present in serum from bovine and swine convalescent from FMDV A12 Lp ab infection. Thus, the highly immunogenic area between residues 137 and 168 of FMDV VP1 elicited a cross-reactive neutralizing idiotope response conserved amongst several animal species. PMID- 8406566 TI - Specific and natural antibody production during Salmonella typhimurium infection in genetically susceptible and resistant mice. AB - Genetically susceptible (C57BL/6) and resistant (CBA) mice were infected with an avirulent strain of Salmonella typhimurium and studied over a 35-day period for the production of antibodies directed against bacterial antigens including lipopolysaccharide (LPS) (specific antibodies) and antibodies directed against self antigens [natural antibodies (NAb)]. Antibodies directed against LPS and self antigens were detected by enzyme immunoassay (EIA) and those directed against other bacterial antigens by immunoblotting. We found that serum natural antibody titres in C57BL/6 and CBA mice were similar and correlated with the bacterial load in the spleen and liver. In C57BL/6 mice, anti-LPS antibodies remained polyreactive and of the IgM isotype. In contrast, CBA mice, after an early increase in polyreactive IgM anti-LPS antibodies, mounted a specific anti LPS IgG antibody response. The immunoblotting results demonstrated that the IgM polyreactive antibodies in the resistant and susceptible mice recognized bacterial antigens of different molecular weights and that CBA, but not C57BL/6 mice, were able to produce IgG antibodies recognizing bacterial components. Our results suggest that the synthesis of antibodies directed against bacterial antigens and natural antibodies follow, at least partially, distinct pathways, but they do not allow us to determine whether these two antibody populations are produced by the same or distinct B-cell subpopulations. PMID- 8406567 TI - Population movement and fate of autoreactive V beta 6+ T cells in Mls-1a mice. AB - In order to demonstrate the precise fail-safe mechanisms involved in the prevention of autoreactive T-cell functions, we analysed the movement of the population of self-reactive V beta 6+ cells in Mls-1a mice. T cells bearing V beta 6 T-cell receptor (TcR) could be detected in the thymus at birth. They increased in number during the next few days, then decreased and disappeared by 1 week after birth. These cells are autoreactive and capable of eliciting a syngeneic graft-versus-host reaction (GVHR). The autoreactive V beta 6+ cells in the thymus on day 3 were abolished by a previous injection of Mls-expressing syngeneic adult spleen cells, showing that the tolerance-inducing antigens had probably not yet developed in newborn mice. These autoreactive V beta 6+ cells escaping clonal deletion may leave the thymus and become appreciable as their percentages rise in the periphery in mice thymectomized 3 days after birth (d3 ThX). However, the 'autoreactive' T cells seemed to be neither cell cycling nor proliferating even after exogenous antigenic stimulation. The proportion of these peripheralized V beta 6+ cells in an 'anergy' state decreased gradually to a half life of about 50 days in adults, in contrast to the complete deletion in a few days of V beta 6hi cells in the developing thymus. On the other hand, in weanlings the percentage of V beta 6+ T cells was reduced to a half-life of less than 20 days, probably because of the diluting out of these cells by the physiological expansion of the irrelevant T-cell population and probably by an increase of body fluid by a factor of 10. In contrast, V beta 8+ T cells, Mls-1a unrelated, maintained a constant proportion, as in non-thymectomized mice. Thus, T-cell repertoire shaping may not always be achieved in the thymus, but may be completed after the cells leave the thymus a few days after birth in a developmentally programmed process. PMID- 8406568 TI - Analysis of human T-cell receptor V beta gene usage following immunization to tetanus toxoid in vivo. AB - The human T-cell antigen receptor (TcR) V beta repertoire was investigated following in vivo reimmunization with tetanus toxoid (TT). Four healthy subjects were immunized subcutaneously with TT, and 24 samples of peripheral blood T cells were taken at intervals over several weeks and used to generate TcR-C beta chain specific first-strand cDNA. A semi-quantitative assay utilizing the polymerase chain reaction (PCR) was used to measure the amount of 22 different TcR-V beta gene transcripts in the cDNA. A peak increase in the amount of V beta 2, 4, 6, 13.1 and 14 occurred 14 days post-immunization, with each V beta increased in at least two of the four subjects. No obvious changes in the other 17 V beta genes were found. A secondary antibody response to TT occurred in all subjects by day 14. These results show that it is now possible to characterize the in vivo kinetics of the human TcR repertoire following stimulation with a conventional antigen. PMID- 8406569 TI - Contribution of N-acetyl-beta-D-galactosamine-specific lectin to Fc receptor mediated phagocytosis by mouse peritoneal macrophages. AB - The contribution of lectin-like receptors on the cell surface of mouse peritoneal macrophages to the process of phagocytosis of IgG-coated sheep red blood cells (SRBC) through Fc receptors has been investigated. Phagocytosis was activated by conditioned medium containing modified vitamin D3-binding protein (DBP) prepared by the incubation of foetal calf serum (FCS) with lysophosphatidyl-choline treated splenic non-adherent cells. Fc receptor-mediated phagocytosis of opsonized SRBC was specifically inhibited by the addition of N-acetyl-beta-D galactosamine. The binding of modified Gc globulin, human DBP, to peritoneal macrophage was only inhibited by the addition of N-acetyl-beta-D-galactosamine, and was dependent on N-acetyl-beta-D-galactosamine concentration. In the presence of cycloheximide, activated phagocytosis was reduced to control levels. By Scatchard plot analysis of binding studies, the number of Fc receptors of macrophages which were activated by conditioned medium increased 3.6-fold in comparison to that of control macrophages. These findings suggest that lectin like receptors having a specificity to N-acetyl-beta-D-galactosamine are involved in activating the process of Fc receptor-mediated phagocytosis of opsonized SRBC by macrophages, and that modified DBP promotes the synthesis of Fc receptors through the N-acetyl-beta-D-galactosamine-specific lectin on macrophage surface. PMID- 8406570 TI - CD44 is not directly involved in the binding of lymphocytes to cultured high endothelial cells from peripheral lymph nodes. AB - The Ager assay was adapted to a porcine lymphocyte-rat high endothelial cell (HEC) system. Using this in vitro assay, the role of porcine CD44 in lymphocyte binding to HEC was examined. The results show that the presence of soluble CD44 molecules did not inhibit the binding of porcine lymphocytes to the cultured rat HEC. Treatment of lymphocytes with anti-CD44 monoclonal antibodies (mAb), or with papain, which removes a 45,000 MW peptide from the intact CD44 molecule, did not inhibit the binding. Binding to the rat HEC did not induce modulation of CD44 molecules on the cell surface. Furthermore, modulation of the CD44 molecule by biotinylated anti-CD44 antibody followed by streptavidin-phycoerythrin, which had caused the molecule to cap on the cell surface, did not prevent the cells binding to the HEC. Similarly, cells denuded of CD44 by anti-CD44 antibody retained the capacity to bind to HEC. Moreover, the binding cells were mainly those which had been stripped of CD44 by the antigenic modulation. It is concluded that CD44 is not directly involved in the binding of lymphocytes to the cultured HEC from peripheral lymph nodes (PLN). PMID- 8406571 TI - Very late antigen integrins are involved in the adhesive interaction of lymphoid cells to human gingival fibroblasts. AB - To date, it is still unclear how the trafficking and retention of activated lymphocytes in periodontal lesions are regulated. In this study, we investigated the molecular basis for the adhesive interactions between lymphocytes and human gingival fibroblasts (HGF). Peripheral blood T lymphocytes (PBT) exhibited binding ability, but only when the calls were activated with phorbol 12-myristate 13-acetate (PMA). Among several human cell lines tested, PMA-stimulated Molt-4, a human T-cell leukaemia line, also displayed significant binding ability to HGF. In order to clarify the molecule(s) involved in this cell-cell interaction, a panel of monoclonal antibodies (mAb) was prepared to PMA-activated Molt-4 and one clone, 4-145, was selected on the basis of its ability to block the binding of PMA-activated Molt-4 to HGF. Moreover, 4-145 inhibited the binding of not only activated Molt-4 but also activated PBT and other cell types to HGF. Biochemical and flow cytometric analyses revealed that 4-145 probably recognizes the beta 1 chain of very late antigen (VLA) integrins. Blocking experiments using mAb specific for the alpha-chain of VLA integrins demonstrated the involvement of alpha 4 (VLA-4) and, to a lesser extent, alpha 5 (VLA-5) chains in the adhesive interactions between T cells and HGF. Despite the significant involvement of VLA integrins in the adhesive interaction between PBT and HGF, the binding of PBT to human dermal fibroblasts (HDF) was not abrogated by 4-145, suggesting that HGF and HDF differ in their requirement of VLA integrins for adhesion to activated PBT. Furthermore, the fact that vascular cell adhesion molecule-1 (VCAM-1), one of the ligands of VLA-4, was not detected on HGF by flow cytometry and anti fibronectin (FN) Ab did not block the adhesive interaction to HGF suggests that not-yet-identified ligand(s) for VLA-4 might be present on HGF. PMID- 8406572 TI - Interleukin-5 is necessary for eosinophilia induced by cyclophosphamide in immunized mice. AB - Interleukin-5 (IL-5) has an important role in the induction of eosinophilia, which is associated with parasitic infestations and with allergic conditions, and which can be induced in a number of experimental systems. One of these model systems involves the administration of cyclophosphamide (CY) to immunized animals. In order to assess the role of IL-5 in this model, eosinophilia was induced in vivo and cell suspensions of spleens or lymph nodes were stimulated in vitro. IL-5 protein secretion was detected by bioassay using an IL-5-dependent cell line (T88-m), and mRNA was assessed by reverse transcription and polymerase chain reaction (RT-PCR). The production of IL-5 protein and mRNA were greatly enhanced in the cells from mice given CY with ovalbumin (OVA), compared with mice given either agent alone. IL-5 protein and mRNA were increased both in spleen and in lymph node cells, and in response either to OVA or to polyclonal stimuli. Further evidence for the importance of IL-5 in this model of eosinophilia was provided by experiments with monoclonal antibodies (mAb) in vivo. A single injection of an IL-5-specific mAb at the time of immunization completely abolished the eosinophilia. By contrast, a monoclonal antibody to IL-4 had no effect. These experiments indicate that IL-5 is required for the eosinophilia induced by CY in immunized mice. PMID- 8406573 TI - Transforming growth factor-beta 1 enhances the generation of allospecific cytotoxic T lymphocytes. AB - We investigated the effects of transforming growth factor-beta 1 (TGF-beta 1) on the proliferation and generation of murine T lymphocytes in vitro. TGF-beta 1 suppressed T- and B-lymphocyte proliferation, mixed lymphocyte reaction (MLR), and the generation of natural killer (NK) cells and lymphokine-activated killer (LAK) cells. On the other hand, TGF-beta 1 significantly enhanced the generation of allospecific cytotoxic T lymphocytes (CTL) at low concentrations (0.01-1 ng/ml) in a dose-dependent manner and restored it to the control level at higher concentrations (10-40 ng/ml). Allospecific CTL generation by TGF-beta 1 was maximally enhanced when added at the beginning of culture. Less enhancement occurred when the addition was delayed. Anti-TGF-beta 1 antibody completely abolished the enhancing effects of TGF-beta 1. Furthermore, platelet-derived TGF beta (pTGF-beta) as well as recombinant TGF-beta 1 similarly enhanced the generation of allospecific CTL. These data demonstrate that TGF-beta has not only immunosuppressive effects but also immuno-enhancing effects in vitro. PMID- 8406574 TI - Characterization of the ovine interleukin-2 receptor-alpha chain: differential induction on precultured alpha beta and gamma delta T cells. AB - A combination of concanavalin A (Con A)-stimulated ovine lymph node (LN) cells and Chinese hamster ovarian (CHO) cells stably transfected with the ovine interleukin-2 receptor-alpha (IL-2R alpha) chain cDNA (CHO IL-2R cells) were used in a differential immunization strategy to generate several monoclonal antibodies (mAb) against the ovine IL-2R alpha chain. The specificity of one of the mAb, designated mAb 9-14, for the ovine IL-2R alpha chain was demonstrated by its reactivity with Con A-stimulated LN cells and CHO IL-2R cells, immunoprecipitation of a 47,000 MW protein from CHO IL-2R cells and inhibition of IL-2-dependent proliferation of Con A-stimulated ovine LN cells. Examination of IL-2R alpha chain expression on resting lamb peripheral blood lymphocyte populations showed a high frequency of IL-2R alpha chain expression on CD4 T cells but not on CD8 T cells, CD45RA+ cells or gamma delta T cells, which comprise up to 60% of lamb peripheral blood T cells. The kinetics of IL-2R alpha chain induction on Con A-stimulated peripheral blood alpha beta and gamma delta T cells was compared. A rapid induction of IL-2R alpha chain expression on precultured gamma delta T cells but not alpha beta T cells was observed within 6 hr of Con A stimulation. A preculturing period was required to 'prime' gamma delta T cells for rapid responsiveness to Con A. Using appropriate inhibitors, we demonstrated that both transcription and translation events were required for rapid IL-2R expression on precultured gamma delta T cells and therefore the 'priming' of gamma delta T cells by in vitro culture did not involve an accumulation of IL-2R alpha chain mRNA or preformed receptors within these cells. PMID- 8406575 TI - The release of transforming growth factor-beta following haemorrhage: its role as a mediator of host immunosuppression. AB - Haemorrhage in the absence of trauma is reported to induce a profound depression in cell-mediated immunity. Recent studies have drawn attention to the cytokine transforming growth factor-beta (TGF-beta) that, while important in wound healing, also has marked immunosuppressive effects. The aim of this study was to determine whether: (1) haemorrhage induces an increase in circulating TGF-beta and if this is associated with the loss of host immunoresponsiveness; and (2) administration of monoclonal antibody (mAb) to TGF-beta following haemorrhage ablates these changes. To determine this, C3H/HeN mice were bled to and maintained at a mean arterial pressure of 35 mmHg for 1 hr. This required removing approximately 50% of the circulating blood volume. Following this period of hypotension, the mice were adequately resuscitated. Blood samples obtained at 24 and 72 hr, but not at 2 hr, following haemorrhage showed a significant elevation in plasma TGF-beta levels when compared to shams. At 24 hr, the increase of TGF-beta in the plasma was associated with decreases in both concanavalin A (Con A)-induced splenocyte proliferation and splenic macrophage antigen presentation. Treating animals with neutralizing antibody (animals received 200 micrograms mAb against bovine TGF-beta 1,2,3/mouse intraarterially) not only reduced the levels of TGF-beta in the blood at 24 hr, but also restored splenocyte functions, such as Con A-induced proliferation, interleukin-2 (IL-2) release, and the capacity of splenic macrophages to present antigen. However, elevated levels of prostaglandin E2 (PGE2) seen in plasma during haemorrhage were only partially depressed by the antibody treatment. These results indicate that the release of TGF-beta contributes to the protracted (> or = 24 hr) suppression of cell-mediated immunity following haemorrhage. PMID- 8406576 TI - Low-affinity receptors for tumour necrosis factor-alpha, interferon-gamma and granulocyte-macrophage colony-stimulating factor are expressed on human placental syncytiotrophoblast. AB - Scatchard binding analysis has been employed to characterize expression of low affinity receptors for tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and granulocyte-macrophage colony-stimulating factor (GM-CSF) by isolated human placental syncytiotrophoblast microvillous plasma membrane (StMPM) vesicles. Trophoblastic receptors for the c-kit ligand (stem cell factor) could not be identified using the same methods. No high-affinity receptors could be detected for GM-CSF or IFN-gamma, but a minority of high-affinity TNF-alpha receptors were identified. Cross-inhibition studies indicated the low-affinity receptors to be specific for each cytokine rather than to be non-specific cytokine-binding factors. Only relatively high-affinity receptors for TNF-alpha and IFN-gamma could be detected on the BeWo human choriocarcinoma cytotrophoblast cell line, whereas receptor affinity for GM-CSF was similar to that on syncytiotrophoblast. Immunohistochemical staining has confirmed expression of IFN gamma receptor by syncytiotrophoblast: in contrast, staining for the established TNF-R1 and TNF-R2 receptors was associated mainly with placental vascular endothelium. These low-affinity cytokine receptors could reflect unique biological responses of foetal syncytiotrophoblast in the presence of local concentrations of maternal cytokine. PMID- 8406577 TI - Glycyrrhizin as a promoter of the late signal transduction for interleukin-2 production by splenic lymphocytes. AB - Cellular and molecular mechanisms of the immunomodulatory action of glycyrrhizin (GL) were studied. We demonstrated that GL displays a unique action to prolong the duration of the T-cell receptor-mediated in vitro splenic T-lymphocyte growth response to anti-CD3 monoclonal antibody (mAb) or concanavalin A (Con A) through enhancement of interleukin-2 (IL-2) secretion and IL-2 receptor (IL-2R) expression. The augmentation by GL of IL-2 production was also found in spleen cells stimulated with A23187 plus phorbol 12-myristate 13-acetate (PMA), suggesting that GL primarily affects some post-receptor stage of the signal transduction. We also found that the time of GL action for promoting IL-2 production and growth response was 2 hr or more after receptor activation. Correspondingly, GL did not augment the anti-CD3 mAb or Con A-mediated protein tyrosine phosphorylation and c-fos transcription. We concluded from these results that GL acts as a promoter of the late signal transduction of T lymphocytes for IL-2 production. PMID- 8406578 TI - Phospholipase C activation in the cytotoxic response of human natural killer cells requires protein-tyrosine kinase activity. AB - Treatment of highly purified natural killer (NK) cells with the protein-tyrosine kinase (PTK) inhibitors, genistein and herbimycin A, diminished their ability to lyse K562 target cells by as much as 100%. The ability of NK cells to bind to K562 cells was not affected by PTK inhibition. However, activation of phospholipase C (PLC) in response to K562 cell binding (as measured by inositol phosphate turnover) was decreased by as much as 75% when PTK activity was inhibited. Furthermore, there was an increase in tyrosine phosphorylation of NK cell PLC gamma 2 after exposure to K562 target cells. These data indicate that a PTK is involved in the activation of NK PLC by tumour target cells in the cytotoxic response. PMID- 8406579 TI - Anti-leucocyte function-associated antigen-1 antibodies inhibit T-cell activation following low-avidity and adhesion-independent interactions. AB - Anti-leucocyte function-associated antigen-1 (LFA-1) antibodies can provide either stimulatory or inhibitory signals to T cells, depending on the epitope they recognize, type and stage of activation of the T cells, and nature of the activation stimulus. Because of the low affinity of interaction between the T cell receptor (TcR) and the antigen/major histocompatibility complex (MHC), it was proposed that the LFA-1 molecule strengthens the adhesion between the interacting cells, thus contributing in an additive manner to TcR-specific interactions. To check if high-avidity, TcR-specific interactions still require the accessory function of the adhesion molecule, we studied the effect of anti LFA-1 antibodies on T-cell triggering mediated through chimeric receptors composed of an Fv of an antibody and a constant region of the TcR. Such chimeric TcR (cTcR) confer on T cells antibody-type specificity and affinity. We made use of transfected T-cell hybridomas expressing various amounts of either one cTcR chain (composed of VH linked to C beta) or double-chain cTcR (VHC beta + VLC alpha). When such transfectants were stimulated with hapten-modified cells, anti LFA-1 antibodies inhibited activation predominantly mediated through cTcR composed of a single chimeric chain and did not inhibit stimulation of the double chain transfectants. Moreover, these anti-LFA-1 antibodies blocked antigen specific T-cell activation regardless of whether the stimulus was adhesion dependent or not, such as in the case of stimulation by immobilized hapten protein conjugates. These studies show that the 'off-signal' provided by anti-LFA 1 antibodies is adhesion independent and affects mainly low-avidity TcR-antigen interactions. PMID- 8406580 TI - Loss of Th1-associated function in peripheral T cells but not thymocytes in tolerance to major histocompatibility complex alloantigen. AB - Mice of the strains A.TH and A.TL were rendered neonatally tolerant to class II major histocompatibility complex (MHC) by the injection of (A.TH x A.TL)F1 spleen and bone marrow cells within 24 hr of birth. Spleen and thymus cells from adult tolerant mice bearing long-term surviving skin grafts were compared with those from normal mice for their in vitro reactivity towards the tolerogen. In a primary mixed lymphocyte reaction (MLR), spleen cells from normal mice proliferated in response to 'tolerogen', generated cytotoxic cells and produced interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) but no IL-4 or IL-5. In contrast, although spleen cells from tolerant mice proliferated and produced IL 2, they failed to generate cytotoxic cells or produce IFN-gamma but produced large amounts of IL-4 and IL-5. The loss of the ability of tolerant cells to generate cytotoxicity or IFN-gamma was profound in that no activity was detected in a secondary MLR and mRNA for IFN-gamma could not be detected by reverse transcription polymerase chain reaction (RT-PCR). To see whether the alteration in function occurred centrally or peripherally, thymus cells from normal and tolerant mice were tested for function. Normal thymocytes produced IFN-gamma, IL 4 and IL-5 in a primary MLR and generated cytotoxic cells in a secondary MLR. In contrast to spleen cells, thymus cells from tolerant mice retained their ability to generate IFN-gamma or cytotoxic cells in response to tolerogen. Overall the results point to a profound switch in peripheral tolerogen-specific responses from a Th 1-biased response in normal mice to a Th2-biased response in tolerant mice and suggest that the alteration in function is post thymic. PMID- 8406581 TI - Generation of rat Th2-like cells in vitro is interleukin-4-dependent and inhibited by interferon-gamma. AB - Differentiation of naive T cells into effector cells producing T helper type 1 (Th1) and Th2 cytokines is regulated by the presence of specific cytokines in the T-cell microenvironment. The effect of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) on Th1- and Th2-like cell development was investigated in cultures of mixed rat spleen cells. These cells were cultured for 4 days in medium containing concanavalin A (Con A) with or without additional IL-2, IFN gamma or IL-4. The cells were then washed and their capacity to produce IL-4, IL 5 and IFN-gamma determined following stimulation with phorbol 12-myristate 13 acetate (PMA) and ionomycin. Freshly isolated cells stimulated with PMA and ionomycin expressed detectable levels of IL-4 and IL-5 mRNA as measured by a quantitative polymerase chain reaction (PCR) procedure and much higher levels of IFN-gamma mRNA. Cells cultured with Con A for 4 days, washed, and restimulated with PMA + ionomycin were unable to express detectable levels of IL-4 and IL-5 mRNA, but produced high levels of IFN-gamma mRNA. Addition of IL-4, or anti-IFN gamma antibody, to Con A-driven splenocyte cultures restored the ability of restimulated cells to express IL-4 and IL-5. CD4+ T cells isolated from these cultures also showed an increased capacity to secrete IL-4 and IL-5 when anti-IFN gamma and IL-4 were present in the culture medium. When cultured for 4 days with Con-A, IL-4 and anti-IFN-gamma splenocytes showed an increased capacity to proliferate in response to recombinant IL-2 and proliferated in response to IL-4 alone. IL-2 had no effect on cytokine production by cultured splenocytes. These results indicate that: (1) IL-4 is essential for the generation of Th2-like cells; (2) IFN-gamma inhibits IL-4 production by mixed spleen cells and suppresses generation of IL-4 responsive T cells; (3) in mixed spleen cell cultures mitogenic stimulation favours differentiation of naive rat T cells into effector cells expressing a Th1, and not Th2, cytokine profile. PMID- 8406582 TI - Differentiation of membrane IgE+ rat B cells into IgE-secreting cells. AB - Rat spleen cells were stimulated with pokeweed mitogen (PWM) and the IgM and IgE responses were assessed. An enrichment of the cell suspension with IgE-bearing cells before stimulation resulted in an increase in the number of IgE-secreting cells. A decrease of the number of IgE-secreting cells was found after depletion of IgE- or IgM-bearing cells, but not those bearing IgD molecules on their membranes, before stimulation. Moreover, the stimulation of membrane IgE on B cells with anti-IgE antibodies was shown to increase the number of IgE-secreting cells after PWM-induced differentiation in vitro. In vivo, it was also observed that a single injection of anti-IgE antibodies can induce the differentiation of IgE-secreting cells. These results demonstrate the presence of IgE(+)-IgM (+)-IgD B cells in the rat that are responsive to PWM-induced differentiation into IgE secreting cells. They indicate a pre-commitment of these cells at a stage where they still express IgM on their surface. IgE molecules on the cell membranes play a role in their differentiation. PMID- 8406583 TI - Brequinar sodium inhibits interleukin-6-induced differentiation of a human B-cell line into IgM-secreting plasma cells. AB - Brequinar sodium (BQR) has been shown recently to be a potent immunosuppressive agent. This property has been attributed to the capacity of BQR to inhibit de novo pyrimidine nucleoside biosynthesis and consequently, to blockade the synthesis both of DNA and RNA. The influence of this new immunosuppressant on lymphocyte function has not been fully characterized. To determine the potential efficacy of BQR for the control of antibody-mediated graft rejection, which is of particular significance in the context of xenotransplantation, we have examined the influence of the drug on interleukin-6-dependent IgM production by the human B-cell line, SKW 6.4. At concentrations up to 10 micrograms/ml, BQR did not affect concanavalin A (Con A)-induced human peripheral blood lymphocyte proliferation or IL-6 production by blood mononuclear leucocytes. In contrast, the drug was very effective in inhibiting IL-6-stimulated IgM production by SKW 6.4 cells, with an optimal inhibitory concentration of 0.3 microgram/ml. As expected, addition of exogenous uridine (0.1 mM), the precursor of uridine triphosphate (UTP), reversed the inhibitory effect of BQR on antibody production, while cytidine (0.1 mM) potentiated the inhibitory activity of the drug. It was further demonstrated that the inhibition of IgM production was unrelated to DNA synthesis, indicating that BQR may affect IL-6 signal transduction and IgM production in SKW 6.4 cells independent of any effect on cell proliferation. PMID- 8406584 TI - Inhibition of interferon-gamma by an interferon-gamma receptor immunoadhesin. AB - Interferon-gamma (IFN-gamma) is an important cytokine which regulates inflammatory and immune response mechanisms. IFN-gamma enhances the presentation and recognition of antigens by inducing the expression of major histocompatibility complex (MHC) proteins, by activating effector T cells and mononuclear phagocytes, and by modulating immunoglobulin production and class selection in B cells. Inappropriate production of IFN-gamma has been implicated in the pathogenesis of several autoimmune and inflammatory diseases and in graft rejection. Here, we describe a recombinant inhibitor of IFN-gamma, termed murine IFN-gamma receptor immunoadhesin (mIFN-gamma R-IgG). We constructed this immunoadhesin by linking the extracellular portion of the mouse IFN-gamma R to the hinge and Fc region of an IgG1 heavy chain. Murine IFN-gamma R-IgG is secreted by transfected cells as a disulphide-bonded homodimer which binds IFN gamma bivalently, with high affinity and in a species-specific manner. In vitro, mIFN-gamma R-IgG can block mIFN-gamma-induced antiviral activity and expression of the class I MHC antigen H-2Kk in cultured cells. In vivo, mIFN-gamma R-IgG can block the function of endogenous mIFN-gamma in mouse models of infection with Listeria monocytogenes and of contact sensitivity. These results show that mIFN gamma R-IgG is an effective and specific inhibitor of mIFN-gamma both in vitro and in vivo. Thus, in general, IFN-gamma receptor immunoadhesins may be useful for investigating the biological functions of IFN-gamma as well as for preventing deleterious effects of IFN-gamma in human disease. PMID- 8406585 TI - Interleukin-8 primes human neutrophils for enhanced superoxide anion production. AB - Interleukin-8 (IL-8), a novel chemotactic cytokine, has been shown to play an important role in inflammation. In this study, we investigated the effect of recombinant human (rh) IL-8 on superoxide (O2-) production by neutrophils. We found that rhIL-8 (1-10 ng/ml) did not stimulate neutrophil O2- production on its own, but primed neutrophils for an enhanced response to other stimuli, such as N formyl-methionyl-leucyl-phenylalanine (FMLP), phorbol 12-myristate 13-acetate (PMA) and platelet-activating factor (PAF). The priming effect of rhIL-8 was dose dependent, rapid and long lasting. Recombinant human IL-8 increased both the maximal rate and the total O2- production, but did not prolong the response to FMLP. Stimulation of neutrophils with rhIL-8 increased intracellular-free calcium concentration ([Ca2+]i) by mobilizing calcium from internal stores and by increasing calcium influx. The increase in [Ca2+]i was dose dependent and occurred in the same range of rhIL-8 concentrations that primed neutrophils for O2- production. In addition, rhIL-8 enhanced the FMLP-stimulated increase in [Ca2+]i. These observations suggest that calcium may play an important role in priming phenomenon. PMID- 8406586 TI - B7/BB-1 is a leucocyte differentiation antigen on human dendritic cells induced by activation. AB - Activation of a primary T-lymphocyte response requires additional signals apart from interaction of the T-cell receptor (TcR)/CD3 complex with major histocompatibility complex (MHC) antigens on the antigen-presenting cell. The CD28 antigen on T lymphocytes provides an important co-stimulatory signal to T lymphocytes and we therefore searched for the presence of its ligand, the B7/BB-1 antigen, on blood and tonsil dendritic cells (DC). Blood DC, prepared from peripheral blood mononuclear cells with a minimal period of in vitro culture, did not stain with the monoclonal antibody BB-1 using flow cytometry analysis. In contrast, tonsil DC stained weakly for B7/BB-1 compared to positive control cell lines. Polymerase chain reaction (PCR) was used to amplify a 605 base pair (bp) fragment from human B7/BB-1 mRNA and demonstrated significant amounts of B7/BB-1 mRNA in tonsil DC but no specific product was obtained from blood DC, confirming the surface-staining results. Weak expression of B7/BB-1 antigen was detected by immunofluorescence analysis following culture of blood DC with either interferon gamma (IFN-gamma) or granulocyte-macrophage colony-stimulating factor (GM-CSF). These data support the concept that blood DC give rise to tissue and/or lymphoid DC, which acquire co-stimulatory ligands as a result of activation and/or differentiation. PMID- 8406587 TI - Protein kinase C transduces the signal for Langerhans' cell migration from the epidermis. AB - Langerhans' cells (LC) take up antigen in the epidermis and then migrate to the local lymph nodes where they present the antigen to T lymphocytes, initiating cutaneous immune responses. However the intracellular mechanisms which mediate LC migration from the epidermis are unknown. We have demonstrated that activation of protein kinase C (PKC) induces this LC migration. An analogue of diacylglycerol (DAG), the physiological activator of PKC, L-alpha-dioctanoyl glycerol (oDAG), applied topically onto the skin of mice caused a significant depletion in the density of Ia+ and J11d+ epidermal LC. oDAG decreased the density of LC in both BALB/c and C57BL mice 24 hr following application, over a dose range of 0.5-24 microM; 200 or 0.05 microM being ineffective. LC density remained depressed for up to 7 days and oDAG increased the number of fluorescein isothiocyanate (FITC) positive cells in the local lymph nodes of mice treated topically with FITC, indicating that oDAG induced LC migration from the epidermis. Additionally, LC migration from the epidermis induced by the contact sensitizer 2,4,6 trinitrochlorobenzene (TNCB) was blocked when PKC was inhibited by palmitoyl-DL carnitine chloride (PCC) or D-Sphingosine (Sph), indicating that LC cannot migrate from the epidermis when PKC is inhibited. However, PCC did not inhibit the induction of contact sensitivity. Thus PKC transduces the signal which leads to LC migration from the epidermis, and disruptions in this secondary messenger may interfere with the induction of immune responses in the skin by disturbing LC migration from the epidermis to the local lymph nodes. PMID- 8406588 TI - Requirements for Langerhans' cell depletion following in vitro exposure of murine skin to ultraviolet-B. AB - Langerhans' cells found within the skin and mucous membranes are critical regulators of antimicrobial and allergic responses. Therefore, the depletion of these cells following exposure of skin to solar ultraviolet radiation (UV) has direct functional consequences on immunity within this tissue. In order to understand how Langerhans' cell depletion is regulated following exposure of skin to medium-wave UV (UVB), the role of second messengers in these responses was investigated using a novel in vitro system. This was accomplished by analysing the expression of a specific marker associated with Langerhans' cells (ATPase) among the epidermal portion of cultured sections of mouse skin following treatment with inhibitors specific for second messenger components and subsequent exposure to UVB. In this study, inhibitors of guanosine triphosphate (GTP) binding proteins, H-8, pertussis toxin and cholera toxin as well as inhibitors of RNA and protein synthesis were all capable of blocking Langerhans' cell depletion in response to UVB treatment. In contrast, an inhibitor of protein kinase C (H-7) was incapable of specifically blocking depletion following treatment with this physical agent. These results suggest that Langerhans' cell depletion mediated by UVB may involve a pertussis and cholera toxin-sensitive G protein as well as de novo protein synthesis. PMID- 8406589 TI - Characterization of C3a anaphylatoxin receptor on guinea-pig macrophages. AB - We have characterized a C3a receptor on guinea-pig macrophages by 125I-C3a binding and functional responses. Scatchard analysis applied to the 125I-C3a binding to guinea-pig macrophages revealed the existence of two receptor classes; a high-affinity class with approximately 0.63 x 10(5) binding sites/cell with a Kd = 2.7 nM, and a relatively low-affinity class with approximately 1.2 x 10(5) binding sites/cell with a Kd = 51 nM. The binding of C3a to macrophages was totally blocked when there was an excess of C3a. C3a triggered a transient intracellular Ca2+ ([Ca2+]i) mobilization in macrophages, which was accompanied by homologous desensitization. C3a was also capable of generating O2- from macrophages. The C3a-induced Ca2+ response and O2- generation were not detected in the pertussis toxin-treated macrophages, suggesting that G proteins are coupled with the C3a receptors of macrophages. Although the C3a-induced O2- generation was inhibited by staurosporine, it was more resistant to staurosporine than phorbol 12-myristate-13-acetate (PMA)-induced O2- generation, suggesting that a protein kinase distinct from protein kinase C may be associated with the C3a receptor. PMID- 8406590 TI - Herpes simplex virus glycoprotein C: molecular mimicry of complement regulatory proteins by a viral protein. AB - Herpes simplex virus (HSV) encodes a protein, glycoprotein C (gC), which binds to the third complement component, the central mediator of complement activation. In this study the structural and functional relationships of gC from HSV type 1 (HSV 1) and known human complement regulatory proteins factor H, properdin, factor B, complement receptor 1 (CR1) and 2 (CR2) were investigated. The interaction of gC with C3b was studied using purified complement components, synthetic peptides, antisera against different C3 fragments and anti-C3 monoclonal antibodies (mAb) with known inhibitory effects on C3-ligand interactions. All the mAb that inhibited gC/C3b interactions, in a differential manner, also prevented binding of C3 fragments to factors H, B, CR1 or CR2. No blocking was observed with synthetic peptides representing different C3 regions or with factor B and C3d, whereas C3b, C3c and factor H were inhibitory, as well as purified gC. There was no binding of gC to cobra venom factor (CVF), a C3c-like fragment derived from cobra gland. Purified gC bound to iC3, iC3b and C3c, but failed to bind to C3d. Glycoprotein C bound only weakly to iC3 derived from bovine and porcine plasma, thus indicating a preference of the viral protein for the appropriate host. Binding of gC was also observed to proteolytic C3 fragments, especially to the beta-chain, thus suggesting the importance of the C3 region as a binding site. Purified gC from HSV-1, but not HSV-2, inhibited the binding of factor H and properdin but not of CR1 to C3b. The binding of iC3b to CR2, a molecule involved in B-cell activation and binding of the Epstein-Barr virus, was also inhibited by the HSV-1 protein. As factor H and properdin, the binding of which was inhibited by gC, are important regulators of the alternative complement pathway, these data further support a role of gC in the evasion of HSV from a major first-line host defence mechanism, i.e. the complement system. In addition, the inhibition of the C3/CR2 interaction may suggest a possible immunoregulatory role of HSV glycoprotein C. PMID- 8406591 TI - Activation of the classical pathway of complement by binding of bovine lactoferrin to unencapsulated Streptococcus agalactiae. AB - The ability of lactoferrin (Lf) bound to Streptococcus agalactiae to interfere with the deposition of complement components on the bacterial surface was investigated by enzyme-linked immunosorbent assay (ELISA). By using a strain of S. agalactiae which activates the alternative pathway of complement in the absence of antibodies, it was found that pretreatment of bacteria with Lf shortened the lag phase preceding the deposition of C3 on bacteria. The kinetics of C3 deposition was comparable to that obtained by adding antibodies against S. agalactiae to agammaglobulinaemic precolostral calf serum (PCS) heated at 56 degrees for 3 min to inactivate the alternative pathway. Accelerated C3 deposition did not occur in the absence of Ca2+ ions. Deposition of C4 on bacteria occurred only when either antibodies or Lf were added to PCS. These results demonstrate that the interaction of lactoferrin with bacteria activated the classical pathway of complement in the absence of antibodies. The binding of purified C1q to bacteria was promoted in a dose-dependent manner by Lf, suggesting that recruitment of classical pathway of complement resulted from the interaction of C1q with Lf adsorbed to the bacterial surface. Phagocytosis of bacteria opsonized with heated PCS (at 56 degrees for 3 min) and Lf was comparable to that occurring in the presence of heated PCS and antibodies. In conclusion, Lf was able to substitute for antibodies in order to activate the classical pathway of complement and to opsonize unencapsulated S. agalactiae efficiently. PMID- 8406592 TI - Anti-phospholipid antibodies in patients with Plasmodium falciparum malaria. AB - Plasma levels of antibodies against phosphatidylinositol (PI), phosphatidylcholine (PC) and cardiolipin (CL) were measured by enzyme-linked immunosorbent assay (ELISA) in patients from malaria endemic area of Sudan and The Gambia. Some Sudanese adults produced IgM antibodies against all three types of phospholipids (PL) during an acute Plasmodium falciparum infection. The anti PL antibody titre returned to preinfection levels in most of the donors 30 days after the disease episode. IgG titres against PI, PC and CL were low. In Gambian children with malaria, IgM antibody titres against PI and PC were significantly higher in those with severe malaria than in those with mild malaria. These results show that a proportion of malaria patients produce anti-PL antibodies during infection and that titres of these antibodies are associated with the severity of disease. PMID- 8406593 TI - Modulation of cellular and humoral immunity, and disease manifestation during onset of patency in Brugia pahangi-infected dogs. AB - Recently, it has become possible to obtain as predicted disease manifestation in selectively bred dogs infected with the naturally occurring lymphatic nematode, Brugia pahangi. In this study, an attempt was made to correlate limb oedema with dynamic changes in immune cell responses occurring in the lymph node at the site of infection during onset of patency. Three litters of dogs were selectively bred; one for the expression of clinical disease, one for the lack of expression of clinical disease and one was of non-specific phenotype. Lymph node cells from 10 of 11 dogs showed a parasite-specific proliferative response at 4-6 weeks post infection (p.i.), before the onset of patency. In six of 11 dogs, a loss of proliferative response to BpA in the infected node cells was detected around the time of onset of patency. In contrast, there was no reduction in the proliferative response to the mitogen phytohaemagglutinin (PHA). The proliferative response to BpA by unresponsive node cells could be restored by addition of substimulatory amounts of murine or human recombinant interleukin-2 (IL-2) to the culture medium. However, there was no correlation between the proliferative response of lymph node cells from infected limbs and the expression of clinical disease. Similarly, when in vitro parasite-specific antibody production by infected lymph node cells was examined, antibody production manifested by all dogs at 5 weeks p.i. was markedly changed at 8 weeks p.i., but these changes did not correlate with clinical disease. This lack of correlation indicates that the immune response to lymphatic filarial infection, as measured in this study, does not necessarily result directly in disease manifestation, and that other genetically determined factors may influence both the parasite specific immune response and the clinical outcome of infection. PMID- 8406594 TI - Characterization of a monoclonal antibody to cis-urocanic acid: detection of cis urocanic acid in the serum of irradiated mice by immunoassay. AB - Cis-urocanic acid (cis-UCA), which is formed from the naturally occurring trans isomer on ultraviolet (UV) irradiation, has been suggested as a photoreceptor for and mediator of the suppressive effects of UV irradiation on systemic immune responses. Trans-UCA is located predominantly in the stratum corneum, and the extent of isomerization to cis-UCA may be analysed by high-performance liquid chromatography (HPLC) of skin extracts. Such an analysis is not suitable for other tissues. In this study a murine monoclonal antibody to cis-UCA was prepared and tested by ELISA using UCA isomers conjugated to protein as antigens. The interaction of the antibody with structural analogues of UCA was assessed by competitive inhibition ELISA which indicated that the antibody had a high specificity for cis-UCA. Screening of sera at various times after UVB irradiation of mice by competitive inhibition ELISA using the monoclonal antibody showed that cis-UCA was present, probably in an unbound form, for at least 2 days after the exposure. Thus, cis-UCA produced in the epidermis following UVB irradiation reaches the serum a few hours later. The implications of this finding for the generation of suppressed immune responses are discussed. PMID- 8406595 TI - Anti-idiotypic immunization provides protection against lethal endotoxaemia in BALB/c mice. AB - Against lipid A (the conserved moiety of lipopolysaccharides from Gram-negative bacteria) neutralizing IgM monoclonal antibodies (mAb) 8-2 and 26-20 anti idiotypic (Ab2) mAb were produced: Ab2 mAb KM-04 (IgG1) against mAb 8-2, and Ab2 mAb PW-1 (IgG2a) and PW-2 (IgG1) against mAb 26-20. The binding of Ab2 mAb KM-04 to 8-2 (Ab1) was strongly inhibited by a lipopolysaccharide (LPS) extract from either Salmonella minnesota R595 (Re LPS) or Escherichia coli J5 (Rc LPS), whereas the binding of Ab2 mAb PW-1 and PW-2 to 26-20 (Ab1) was only marginally inhibited by both Re LPS and Rc LPS. The results indicated that Ab2 mAb KM-04 recognizes a lipid A-binding site related idiotope on mAb 8-2 and therefore KM-04 might bear the internal image of a neutralization determining epitope of lipid A. Consequently Ab2 KM-04 might induce antibodies to lipid A. Indeed anti-idiotypic immunization of syngeneic BALB/c mice with Ab2 mAb KM-04 resulted in development of lipid A-binding anti-anti-idiotypic (Ab3) antibodies in serum. Similar immunizations with Ab2 mAb PW-1 and PW-2 were unsuccessful. However, induction of lipid A-binding Ab3 by mAb KM-04 proved to be genetically restricted to BALB/c mice. DBA/2 mice, Swiss mice and rabbits did not develop lipid A-binding antibodies upon immunization with mAb KM-04. In protection experiments, it was shown that BALB/c mice vaccinated with mAb KM-04 showed significantly enhanced survival from challenge with either rough (Re) LPS from Salmonella minnesota or smooth LPS from E. coli 0111:B4 when compared to BALB/c mice immunized with a non relevant Ab2 mAb. The results suggest that mAb KM-04 constitutes a non-internal image vaccine to the lethal effect of lipid A in BALB/c mice. Furthermore an Ab3 mAb was prepared against Ab2 mAb KM-04 that showed reactivity with Re LPS. This Ab3 mAb, designated LE-21 (IgG2a) protected mice against an otherwise lethal challenge of Re LPS. PMID- 8406596 TI - Polyclonal anti-idiotypes induce antibody responses protective against ricin cytotoxicity. AB - Protein G-purified goat anti-ricin IgG, previously demonstrated to protect against ricin toxicity in vitro and in vivo, was used to raise BALB/c mouse and New Zealand White rabbit polyclonal anti-idiotypic antibodies. The generated anti idiotypic sera were repeatedly absorbed over agarose conjugated to normal goat immunoglobulins, and purified by protein A-agarose affinity chromatography. Immunization of BALB/c mice with BALB/c anti-idiotypes did not result in a significant anti-ricin antibody response. However, injection of BALB/c mice with BALB/c anti-idiotypes conjugated to keyhole limpet haemocyanin (KLH) or with unconjugated rabbit anti-idiotypes resulted in specific and anti-ricin immune responses. The anti-idiotype-induced anti-ricin antibody responses protected against the in vitro cytotoxicity of ricin, a potent plant-derived protein synthesis inhibitor, as assessed by the murine EL-4 leukaemia cell assays. Thus, anti-idiotype-based vaccines may represent an alternative, safe and effective means of inducing protective immunity against toxins such as ricin, whose extreme in vivo toxicity render them unsafe as immunogens. PMID- 8406597 TI - Lipid composition of intestinal brush border membrane in alloxan induced acute experimental diabetes. AB - Several alterations were observed in the rat intestinal brush border membrane (BBM) lipid composition after 7 days old alloxane-induced diabetes as compared to the control animals. There was no change in the total protein contents but a significant increase in the total lipid contents was observed. Glycolipids constituting the major lipid components showed a two-fold increase. No significant difference was observed in the total phospholipid contents. A significant decline in the free cholesterol (CH) level, free fatty acids, triglycerides and sialic acid contents was observed in membranes from diabetic rats. Esterified CH, monoglycerides+diglycerides, phosphatidyl serine+phosphatidyl inositol and phosphatidyl choline levels remained unaffected. A significant increase in sphingomyelin with a parallel decrease in phosphatidyl ethanolamine was observed in BBM preparations from diabetic rats. The observed changes in intestinal BBM might be responsible for altered functions of the diabetic intestines. PMID- 8406598 TI - Sodium chloride stress induced morphological and ultrastructural changes in Aspergillus repens. AB - Halotolerant fungus, A. repens, showed a considerable difference in its growth rate, morphology, ultrastructural and molecular composition under NaCl stress as compared to control i.e. non-stressed condition. Light microscopic observations revealed significant differences in their mycelial thickness, their branching and septa. Transmission electron microscopic observations of both the conditions depicted significant differences in the qualitative and quantitative changes in mitochondria. Frequent pinocytotic vesiculation (vacuoles) of plasma membrane was observed in fungus under stress but no such vesiculation in control. The multivesiculate structures observed under stress with their origin from the cell membranes and subsequent release into vacuoles have not been reported in fungi under normal physiological conditions. The observations on pinocytosis are discussed in relation to ion compartmentation and salt tolerance in A. repens. PMID- 8406599 TI - Cellular distribution of hexa and trivalent chromium in Pseudomonas aeruginosa. AB - Binding of Cr to P. aeruginosa whole cells increased with an increase in positive charge of the chromium complexes used. Distribution of Cr bound from aqueous solutions of potassium dichromate--a Cr(VI) salt and basic chromium sulphate (BCS)--a Cr(III) salt [used for tanning which has 25% of Cr2O3 on weight basis and is of 33% basicity (one hydroxyl/Cr atom)] was more in soluble fraction (protein and nucleic acids) than in insoluble fraction (lipids, polyphosphates, polysaccharides, lipopolysaccharides and mucopeptides). Electron spin resonance spectrum of the cells exposed to hexavalent chromium revealed the presence of Cr(III) ion demonstrating a reduction of the same by the Pseudomoas. The infrared spectra of chromium exposed cell envelopes support that protein carboxylic sites and proteins are involved in binding Cr(III) and predominantly lipids in Cr(VI). Bound chromium could be removed chemically and the cells could then be reused as a biosorbent. PMID- 8406600 TI - Effect of pH and sodium ions on intestinal uptake of lysine in rats. AB - Intestinal uptake of lysine in rats progressively decreased with an increase in pH from 5.2 to 8.5, both in the presence and absence of Na+ ions. At pH 5.2 lysine uptake was 30-35% more than that at neutral pH. Na+ activated lysine uptake by 40-50% at pH 5.2 and it was increased to 110-120% at neutral pH. The observed increase in lysine uptake in response to Na+ and H+ gradients was due to enhanced maximal velocity (Vmax), with little change in affinity constant (Kt). Arrhenius analysis revealed a biphasic curve for lysine uptake with transition temperature (Tc) around 20 degrees C (24 degrees C at pH 5.2 in presence of Na+). The energy of activation (Ea) below (16.1-23.4 Kcal/mole) and above (6.7-8.6 Kcal/mole) the Tc was similar at pH 5.2 and 7.0 both in the presence and absence of Na+ ions. The sensitivity of lysine uptake to various inhibitors was also dependent upon pH and Na+ ions. PMID- 8406601 TI - Anti-platelet activating factor property of Rubia cordifolia Linn. AB - Rubia cordifolia is clinically used for the purification of blood by the physicians of the Indian System of Medicine. For the first time, the effect of the partially purified fraction of this whole plant has been studied on rabbit platelets. It inhibits the platelet aggregation induced by PAF (platelet activating factor) but not thrombin. It also inhibits the binding of 3H-PAF to the platelets in the dose-dependent manner. Thus it appears that R. cordifolia inhibits action of PAF at its receptor level either by it's blocking or by desensitization. PMID- 8406602 TI - Effect of high-fat diet on mice intestinal brush border membrane composition. AB - Effect of feeding high-fat (26% fat) diet to mice for 21 days on intestinal brush border membrane composition was evaluated by comparing with controls fed 10% fat diet. 125I-labelled lectin binding and chemical analysis of fucose, sialic acid, hexoses and hexosamines revealed essentially similar results in control and test groups. Membrane phospholipids, expressed on dry membrane basis, were significantly reduced while total cholesterol was enhanced in experimental group compared to controls. Triglyceride content was not altered under these conditions. [14C]-acetate incorporation studies showed that decrease in phospholipid content was due to reduced synthesis of phospholipid constituents, in particular, phosphatidylcholine and phosphatidylethanolamine, as a result of feeding high fat diet. The results suggest that high amount of fat in the diet of adult mice does not alter sugar content in brush border membrane but affects membrane lipid composition. PMID- 8406603 TI - Levels of estrogen and progesterone in milk of animals induced into lactation by reproductive steroids and thyrotropic releasing hormone. PMID- 8406604 TI - In vitro encystation and excystation of Entamoeba histolytica trophozoites. PMID- 8406605 TI - Effect of oral administration of fungicide-mancozeb on thyroid gland of rat. PMID- 8406606 TI - Role of mushroom (Pleurotus florida) as hypocholesterolemic/hypolipidemic agent. PMID- 8406607 TI - Dehydrogenases of pentose phosphate pathway and cervical carcinogenesis. PMID- 8406608 TI - Radiomodifying effect of camphor as evaluated by micronuclei frequency and sister chromatid exchanges. PMID- 8406609 TI - Non-specific effects of Setaria cervi antigen, adjuvants and vaccines against Litomosoides carinii infection in cotton rats, Sigmodon hispidus. PMID- 8406610 TI - Effects of alloxan induced diabetes on lipid profiles in renal cortex and medulla of mature albino rats. PMID- 8406611 TI - Physical location of the human immunoglobulin lambda-like genes, 14.1, 16.1, and 16.2. AB - The human immunoglobulin lambda-like (IGLL) genes, which are homologous to the human immunoglobulin lambda (IGL) light chain genes, are expressed only in pre-B cells and are involved in B cell development. Three IGLL genes, 14.1, 16.1, and 16.2 are present in humans as opposed to one, lambda 5 (Igll), found in the mouse. To precisely map the location of the human IGLL genes in relation to each other and to the human IGL gene locus, at 22q11.1-2, a somatic cell hybrid panel and pulsed field gel electrophoresis (PFGE) were used. Hybridization with a lambda-like gene-specific DNA probe to somatic cell hybrids revealed that these genes reside on 22q11.2 between the breakpoint cluster region (BCR) and the Ewing sarcoma breakpoint at 22q12 and that gene 16.1 was located distal to genes 14.1 and 16.2. Gene 14.1 was found by PFGE to be proximal to 16.2 by at least 30 kilobases (kb). A 210 kb Not I fragment containing genes 14.1 and 16.2 is adjacent to a 400 kb Not I fragment containing the BCR locus, which is just distal to the IGL-C (IGL constant region) genes. We have determined that the IGLL genes 14.1 and 16.2 are approximately 670 kb and 690 to 830 kb distal, respectively, to the 3'-most IGL-C gene in the IGL gene locus, IGL-C7. We thus show the first physical linkage of the IGL and the IGLL genes, 14.1 and 16.2. We discuss the relevance of methylation patterns and CpG islands to expression, and the evolutionary significance of the IGLL gene duplications. Consistent with the GenBank nomenclature, these human IGLL genes will be referred to as IGLL1 (14.1), IGLL2 (16.2), and IGLL3 (16.1), reflecting their position on chromosome 22, as established by this report. PMID- 8406612 TI - Genomic organization and tissue expression of the mouse proteasome gene Lmp-7. AB - LMP7 is one of the two proteasome subunits encoded in the major histocompatibility complex and is speculated to play a role in the generation of endogenous peptides for presentation by class I molecules to cytotoxic T cells. Here we report the genomic organization of the mouse Lmp-7 gene and the tissue distribution of its messenger RNA. In contrast to human LMP7 which is composed of seven exons and six introns, the mouse Lmp-7 gene is organized in six exons and five introns. Interestingly, the region corresponding to the first exon of human LMP7 is highly modified by numerous insertions and deletions and contains two in frame stop codons. Consequently, the mouse Lmp-7 gene does not allow the alternative exon usage described in humans and most likely encodes for only one LMP7 protein. Thus, the Tap-1 3' end gene region and the Lmp-7 initial translation codon are separated by an 1182 nucleotide region which contains a TATA-box, a cAMP regulatory element, two SP1 sites, and two G-C-rich regions. Expression of the Lmp-7 messenger RNA was analyzed on different tissues from unstimulated mice. Lmp-7 messenger RNA is expressed in spleen, thymus, lung, liver, heart, and, at a very low level, in kidney but not in brain and testis. The possible role of Lmp genes in antigen processing is discussed. PMID- 8406613 TI - Zebrafish Mhc class II alpha chain-encoding genes: polymorphism, expression, and function. AB - Its small size and short generation time renders the zebrafish (Brachydanio rerio) an ideal vertebrate for immunological research involving large populations. A prerequisite for this is the identification of the molecules critical for an immune response in this species. In earlier studies, we cloned the zebrafish genes coding for the beta chains of the class I and class II major histocompatibility complex (Mhc) molecules. Here, we describe the cloning of the zebrafish alpha chain-encoding class II gene, which represents the first identification of a class II A gene in teleost fishes. The gene, which is less than 3 kilobases (kb) distant from one of the beta chain-encoding genes, is approximately 1.2 kb long and consists of four exons interrupted by very short (< 200 base pairs) introns. Its organization is similar to that of the mammalian class II A genes, but its sequence differs greatly from the sequence of the latter (36% sequence similarity). Among the most conserved parts is the promoter region, which contains X, Y, and TATA boxes with high sequence similarity to the corresponding mammalian boxes. The observed striking conservation of the promoter region suggests that the regulatory system of the class II genes was established more than 400 million years ago and has, principally, remained the same ever since. Like the DMA, but unlike all other mammalian class II A genes, the zebrafish gene codes for two cysteine residues which might potentially be involved in the formation of a disulfide bond in the alpha 1 domain. The primary transcript of the gene is 1196 nucleotides long and contains 708 nucleotides of coding sequence. The gene is expressed in tissues with a high content of lymphoid/myeloid cells (spleen, pronephros, hepatopancreas, and intestine). The analyzed genomic and cDNA sequences are probably derived from different loci (their overall sequence similarity in the coding region is 73% and their 3' untranslated regions are highly divergent from each other). The genes are apparently functional. Comparison of genes from different zebrafish populations reveals high exon 2 variability concentrated in positions coding for the putative peptide-binding region. Phylogenetic analysis suggests that the zebrafish class II A genes stem from a different ancestor than the mammalian class II A genes and the recently cloned shark class II A gene. PMID- 8406614 TI - Polymorphism of the DQA1 promoter region (QAP) and DRB1, QAP, DQA1, DQB1 haplotypes in systemic lupus erythematosus. SLE Study Group members. AB - We have investigated the DNA polymorphism for the DQA1 promoter region (QAP) and HLA-class II DRB1, DQA1, and DQB1 genes in 178 central European patients with Systemic lupus erythematosus (SLE) using polymerase chain reaction and Dig-ddUTP labeled oligonucleotides. Increased frequencies of DRB1*02 and *03 are confirmed by DNA typing. In addition, the frequencies of DQA1*0501, *0102 and DQB1*0201, *0602 alleles are increased in the patients as compared to controls. The strongest association to SLE is found with DRB1*03 and DOB1*0201 alleles (p < 10( 7), p corr. < 10(-5) and p < 10(-6), p corr. < 10(-4), respectively). By investigating the DQA1 promoter region in the SLE patients we have detected nine different QAP variants. Increased frequencies of QAP1.2 and QAP4.1 are observed in patients as compared to controls (p < 0.05, p corr. = n.s.). Analysis of linkage disequilibria demonstrates a very strong association between QAP variants and DQA1, DRB1 alleles. Certain QAP variants are completely associated with DQA1 and DRB1 alleles, whereas others can combine with different DQA1 and DRB1 alleles. All DRB1*02-positive patients and controls carry QAP1.2, and all DRB1*03 positive patients and controls carry QAP4.1. Conversely, the QAP1.2 variant appears only in DRB1*02 haplotypes, while the QAP4.1 variant can be observed in DRB1*03, *11, and *1303 haplotypes. Based on the strong linkage disequilibria between DRB1-DQA1-DQB1 genes and between DRB1-QAP-DQA1, we have deduced the four point haplotypes for DRB1-QAP-DQA1-DQB1 in patients and controls. Two haplotypes DRB1*02-QAP1.2-DQA1*0102-DQB1*0602 and DRB1*03-QAP4.1-DQA1*0501-DQB1*0201 are significantly increased in patients as compared to controls (p < 0.01, p corr. = n.s., RR = 1.8 and p < 10(-7), p corr. < 10(-5), RR = 3.1, respectively). The analysis of relative risks attributed to the various alleles of QAP, DQA1, and DQB1 as well as the investigation of the deduced DRB1-QAP-DQA1-DQB1 haplotypes leads to the conclusion that QAP4.1 and DQA1*0501 on the DR3 haplotypes are probably not involved in SLE susceptibility.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8406615 TI - Molecular structures of cattle T-cell receptor gamma and delta chains predominantly expressed on peripheral blood lymphocytes. PMID- 8406616 TI - Lack of association with T-cell receptor TCRBV6S1*2 allele in HLA-DQA1*0101 positive Norwegian juvenile chronic arthritis patients. PMID- 8406617 TI - Characterization of HLA-DMB polymorphism. PMID- 8406618 TI - How the ratio of nonsynonymous to synonymous pseudogene substitutions can be less than one. PMID- 8406619 TI - Human CD4 produced in lymphoid cells of transgenic mice binds HIV gp120 and modifies the subsets of mouse T-cell populations. PMID- 8406620 TI - A study of DR2-LUM haplotype generation and the DRB6*0202 linkage to DRB1*1601. PMID- 8406621 TI - Sequence analysis of DPB1-like genes in cynomolgus monkeys (Macaca fascicularis). PMID- 8406622 TI - A novel HLA-DRB1 allele (DRB1*0417) in South American Indians. PMID- 8406623 TI - Isolation of a clone for pig beta 2-microglobulin cDNA. PMID- 8406624 TI - Murine hypersensitivity pneumonitis: evidences for the role of eicosanoids and platelet activating factor. AB - Eicosanoids and platelet activating factor (PAF) are involved in numerous lung diseases. However, few studies have looked for their role in hypersensitivity pneumonitis (HP). HP was induced in mice by repeated instillations of Saccharapolyspora rectivirgula (Micropolyspora faeni) during 3 weeks. Bronchoalveolar lavages (BAL) were performed each week. BAL macrophage, lymphocyte and neutrophil counts increased at week 1, for a maximal increase after 3 weeks. Production of LTB4, LTC4, PGE2 and TXB2 by alveolar macrophages stimulated in vitro with calcium ionophore was maximal after 3 weeks, the appearance of PGE2 and TXB2 preceding that of LTB4 and LTC4. Eicosanoid production by AM correlated with BAL cell counts. To look for the importance of PAF in the development of experimental HP, mice were treated with BN52021 and BN50730, two PAF antagonists, and BAL were performed after 3 weeks of treatment. The compounds BN52021 and BN50730 reduced total BAL cell counts (x 10(5) cells/ml) in animals treated with S. rectivirgula from 8.4 +/- 0.8 to 4.4 +/- 1.1 and 3.7 +/- 0.2 respectively. BAL cell numbers in control animals were 0.4 +/- 0.1. In conclusion, eicosanoids are increased in experimental HP, and, as PAF, could play a role in the accumulation of alveolar cells. PMID- 8406625 TI - Chicken IgY-Fc antibody elicited in a rabbit as first coat on frosted glass beads in immunoassays. AB - The observation of Esteves and Binaghi that the Fc moieties of mammalian IgG are antigenically similar but antigenically unrelated to the Fc fraction of chicken IgG (IgY) was usefully applied in enzyme linked immuno-sorbent assay (ELISA). In this communication its use in ELISA of two plant viruses viz. tobacco mosaic virus (common strain) and grape vine A virus is illustrated. Frosted glass beads were used as reagent carriers and the reactions were conducted in the wells of multi-titre plates. PMID- 8406626 TI - Synovial mononuclear phagocytes in rheumatoid arthritis and osteoarthritis: quantitative and functional aspects. AB - Macrophages are normal constituents of synovial tissue, and in inflammatory synovitis the number of synovial macrophages increases. Synovial macrophages and their secretory products are important in initiating, propagating, and maintaining the synovial inflammation in rheumatoid arthritis (RA). The purpose of this study was to determine the absolute numbers of macrophages in synovia resected from patients with RA and osteoarthritis (OA) and to determine their abilities to produce and/or functionally express tumor necrosis factor (TNF), interleukin-1 (IL-1), and tissue factor (thromboplastin). Results demonstrate that synovial tissue from RA patients (as compared to that from OA patients) weighed more, contained more cells, more macrophages, and more multinucleated giant cells (macrophage polykaryons). Also, isolated cells from both OA and RA patients had tissue factor activity and could produce TNF and IL-1 with in vitro culture, but these parameters were not different in cells from OA and RA patients. RA patients receiving glucocorticoid treatment for their arthritis had fewer total synovial cells than did patients not on glucocorticoids, but treatment with nonsteroidal anti-inflammatory agents did not alter cell numbers. Patient treatment with glucocorticoids or non-steroidal anti-inflammatory drugs did not influence the ability of their isolated cells to produce TNF or IL-1. PMID- 8406627 TI - Modulation of transforming growth factor-beta 1 effects by cytokines. AB - The purpose of the present study was to determine the effects of human recombinant transforming growth factor-beta 1 (TGF-beta 1) on the proliferation of normal cell and cancer cell lines and to evaluate the mechanism of TGF-beta induced immunosuppression. Murine H238 fibrosarcoma and human UC-11 glioblastoma cells showed no proliferative change in the presence of TGF-beta, whereas the growth of human LS174T colon adenocarcinoma cells was significantly enhanced at the lower concentrations of TGF-beta. In contrast, Mono/Mac-6, a human monocyte cell line, human peripheral blood mononuclear (PBMN) cells, and BALB/c mouse spleen cells were significantly suppressed by 2.5 to 250 ng/ml of TGF-beta. In order to investigate the mode of action, TGF-beta and other cytokines were added 0, 1, and 2 days after initiation of the culture. Mono/Mac-6 cells showed that 2 days are needed for TGF-beta-induced suppression. Simultaneous addition of TGF beta and tumor necrosis-alpha (TNF-alpha; 600 units/ml) to Mono/Mac-6 cells resulted in nearly complete suppression by day 3. IL-2, and to a lesser extent IL 4, was able to counteract the suppressive effects of TGF-beta on mitogen stimulated spleen cells. However, our results indicate that IL-2 is not as effective in restoring responsiveness once T cell activation is well underway. IL 1 and interferon-gamma had no effects on TGF-beta-mediated immunosuppression. Since TGF-beta depressed normal cell growth and since IL-2 could effectively counteract the suppression, we assayed for IL-2 production. When normal spleen cells were treated with 2.5 ng of TGF-beta/ml, a 3.4-fold decrease in IL-2 production was observed. This is a potential mechanism for TGF-beta-mediated immunosuppression. PMID- 8406628 TI - Epidermal dendritic cell populations in the flaky skin mutant mouse. AB - Flaky skin (gene symbol: fsn) is an autosomal recessive mouse mutation that causes pathologic changes in the skin yielding a papulosquamous disease resembling human psoriasis. Preliminary studies of epidermal sheets from foot pads of fsn/fsn mice stained for Ia+ Langerhans cells (LC) or Thy-1+ dendritic epidermal cells (Thy-1+ DEC) indicated a rise in LC numbers at the time of weaning, when the skin lesion becomes clinically evident. To further investigate this observation, epidermal sheets were obtained from the ear, dorsal skin, and foot pads from replicates of 6 female mice (both mutants and normal littermates) on weekly intervals from birth to 8 weeks of age. Dorsal skin epidermal thickness was quantitated by computer assisted image analysis and found to be significantly thickened from one week onward in the mutant mice. Using immunofluorescence microscopy, epidermal dendritic cell numbers were determined following staining with antibodies for the following markers: Ia, NLDC-145, and S-100 (for LC) or Thy 1.2 and asialo-GM1 (for Thy-1+ DEC). Use of all 5 markers to evaluate skin from 3 different locations yielded a subtle but significant increase in LC and Thy-1+ DEC in flaky skin mice. Of the three sites evaluated, the dorsal skin and ear epidermal sheets were most informative, which corresponded to the degree of pathological involvement. Mice doubly homozygous for fsn and for the severe combined immunodeficiency (scid) mutation developed the psoriasiform dermatitis. Bone marrow grafts from fsn/fsn homozygotes to homozygous scid/scid mice reproduce the skin lesion. These studies suggest that the psoriasiform dermatitis in the flaky skin mouse mutation is associated with abnormalities at the level of hematopoietic progenitor cells. PMID- 8406629 TI - Prevalence of initial drug resistance in tuberculosis patients attending a chest hospital. AB - Sputum samples from pulmonary tuberculosis patients attending a hospital for chest diseases and tuberculosis at Jaipur, India were directly subjected to sensitivity tests to detect drug resistance to streptomycin (S), isoniazid (I), rifampicin (R) and ethambutol (Emb) by slide culture technique. Drug resistance was observed to one or more drug in 19.9 per cent of the patients. I resistant organisms were present in 10.1 per cent of patients, S resistance in 7.6 per cent, R resistance in 3.0 per cent and Emb resistance in 2.6 per cent. Resistance was limited to a single drug in 16.7 per cent patients. Drug resistance was unrelated to age and sex of the patients. PMID- 8406630 TI - Value of a single Widal test in the diagnosis of typhoid fever. AB - Results of a single Widal test in patients with bacteriologically confirmed typhoid fever (116), clinically suggestive but culture negative fever (170) and non-typhoidal febrile illness (98) and in normal control children (54) were analysed. Positive Widal test (antibody titre against S. typhi O antigen of 1:160) was recorded in 61.2 per cent of patients with bacteriologically confirmed typhoid fever and in 58.8 per cent with culture negative but clinically suggestive typhoid fever. In contrast, the same titre was observed in 10.2 per cent patients with other febrile illnesses of known etiology and in 1.8 per cent of normal children. Differences in the positivity of Widal test in patients with bacteriologically confirmed typhoid fever and clinically suggestive but culture negative fever were highly significant (P < 0.000001) when compared to that of patients with non-typhoidal febrile illnesses and normal controls. High specificity and positive predictive value in 1:160 dilution makes the Widal test acceptable as a diagnostic tool. PMID- 8406631 TI - Shortcomings of some currently available DNA probes for malaria detection. AB - A non-radioactive DNA probe based-method for detecting malaria will greatly aid epidemiological studies. Using putative Plasmodium falciparum and P. vivax specific 18S ribosomal RNA directed oligonucleotides, different enzymatic and chemiluminescent detection methods were attempted without success. The sensitivity of the corresponding 32P-labelled probes was found to be inadequate. A published procedure based on chemiluminescent detection of repetitive DNA sequences of P. falciparum was found to be adequately sensitive but lacking in specificity. PMID- 8406632 TI - Prevalence of filariasis in rural areas of Shahjahanpur district (Uttar Pradesh). AB - Night blood survey was carried out during May, 1990 to December, 1991 in 18 villages of 5 Primary Health Centres of district Shahjahanpur (UP), to find out the prevalence of filariasis in the area. Out of 2141 individuals surveyed randomly, 217 were found positive for microfilariae of Wuchereria bancrofti. The microfilaria rate, filarial disease rate and filarial endemicity rates were 10.1, 11.4 and 18.8 per cent respectively. An entomological survey revealed Culex quinquefasciatus as the vector. The average man hour density was 25.8. It is clear from the results that filariasis is more endemic in rural areas than urban area of Shahjahanpur as observed by local filariasis control unit. PMID- 8406633 TI - Detection of sialic acid in trophozoites of Entamoeba histolytica (NIH:200). AB - In two successive experiments detection of sialic acid was achieved from trophozoites of axenically grown E. histolytica (NIH:200) using silicic acid column chromatography for the separation of sialosylated lipids and asialosylated lipids from total lipids. Sialic acid of the sialosylated lipids was detected through thiobarbituric acid assay followed by acid hydrolysis. These findings indicate that presence of sialic acid is not only a character of the cystic stage but also that of trophozoites of E. histolytica. Detection of sialic acid confirmed the electronegative charge of the surface membrane (glycocalyx) of E. histolytica trophozoites. PMID- 8406634 TI - Evaluation of serological methods for determination of specific IgE in diagnosis of aspergillus lung disease. AB - Forty six patients clinically suspected to have aspergillus lung disease [bronchial asthma 17, allergic broncho-pulmonary aspergillosis (ABPA) 22, aspergilloma 7] were evaluated by Phadebas RAST, Phadezym RAST (ELISA) and avidin biotin ELISA for determination of specific IgE. Smear, culture, skin test against aspergillin, precipitin test and Enzygnost (total IgE) confirmed the diagnosis in 7 patients of extrinsic asthma due to conidia of Aspergillus spp., 17 for ABPA and 7 for aspergilloma among the clinically suspected patients. Specific IgE against Aspergillus fumigatus antigen was detected in 100 per cent patients of extrinsic asthma and 64.7 per cent of ABPA patients. No specific IgE could be detected in patients with aspergilloma. In patients other than those with, aspergilloma the detection rate of specific IgE was 50.0 per cent by radio allergosorbent test (RAST), 62.5 per cent by ELISA, 66.7 per cent by avidin biotin ELISA. Thus ELISA or avidin-biotin ELISA could replace the radioimmunoassay procedure for detection of specific IgE. PMID- 8406635 TI - Vapour toxicity & repellence of some essential oils & terpenoids to adults of Aedes aegypti (L) (Diptera: Culicidae). AB - Bioefficacy of commercially used synthetic insecticides/repellents and potential of selected essential oils and terpenoids were assessed against mosquitoes. Essential oils and terpenoids, were vapourised in commercially manufactured mosquito repellent electronic assemblies and effects of such vapours were tested on 6-7 days old adult female Aedes aegypti. Commercially available 'mats' (coir rectangles) impregnated with allethrin were used as standards for comparison of Kt50 and Kt90 values. Fastest knock-down was seen in case of allethrin, followed by terpeneol (anhydrous) and (-) carvone. Maximum knock-down time was observed for beta citronellol. All compounds exhibited a repellent effect also, terpeneol (anhydrous) being the best, followed by (-) carvone and citronellal. In repellent tests, no mortality was caused by terpenoids, but allethrin caused > 80 per cent knock-down. PMID- 8406636 TI - Persistence of leech repellents on cloth. AB - Trials on persistence of repellent properties of N, N-diethyl phenyl acetamide (DEPA), N,N-diethyl-m-toluamide (DEET), 3acetyl2(2-6-dimethyl-5 heptenyl)oxazolidine(Citronyl) , dimethyl phthalate (DMP) and N-benzoyl piperidine (NBP) on cloth were conducted against land leeches in evergreen rain and deciduous forests of Assam. Results obtained were compared with volatile oil of Zanthoxylum armatum DC. syn. Z. alatum Roxb (Timur) to evaluate its efficacy as leech repellent. DEPA and DEET were found to be the best. Timur oil was at par with Citronyl and exhibited better results than DMP and NBP. PMID- 8406637 TI - Dengue in Gujarat state, India during 1988 & 1989. AB - Following the reports of epidemics of febrile illness from several rural and urban areas of Gujarat state (India) in 1988, epidemiological investigations were carried out and dengue (DEN) virus activity was demonstrated in large cities such as Surat and Rajkot as well as several villages in Sabarkantha district. Two strains of dengue type-2 each were isolated from human sera from Surat city and a village in Sabarkantha district. Six strains of dengue virus were isolated from Aedes aegypti mosquitoes collected at Chotasan village, two of which were confirmed as DEN type-2. Of the 560 patients' sera tested from different areas (including villages and townships), 122 showed evidence of dengue infection and another 236 showed a broader reaction with flaviviruses. Entomological investigations showed a widespread distribution of Ae. aegypti both in urban and rural areas. In the household conditions this mosquito was found to breed predominantly in containers with non-potable water. Amongst these, cement containers manufactured in towns and distributed to the villages seem to play an important role in the spread of this species. In non-residential areas prolific breeding of Ae. aegypti was observed in automobile tyre dumps, and varied types of scrap, in towns and villages. Distribution and relative prevalence of the species were studied in 46 towns and villages, covering the spectrum of rural urban-continuum. These studies provide an indication regarding the mechanism of the spread of DEN virus through peoples' movement, transport, the process of urbanisation etc. PMID- 8406638 TI - Bactericidal action of pulsed exposure to rifampicin, ethambutol, isoniazid & pyrazinamide on Mycobacterium tuberculosis in vitro. AB - The bactericidal action of pulsed exposure to rifampicin (R), ethambutol (Emb), isoniazid (I) and pyrazinamide (Z) together on alternate days (REmbIZ) and as REmb and IZ separately on alternate days (REmb/IZ) on M.tuberculosis H37Rv, two isolates of M.tuberculosis sensitive to these drugs, as well as four isolates resistant to one or more drugs, was studied using an in vitro method. The experimental duration was 6 days. REmbIZ and REmb/IZ appeared to have equally good bactericidal action on M.tuberculosis strains in the in vitro system. The results suggest that splitting REmbIZ into REmb and IZ on alternate days in short course chemotherapy regimens for tuberculosis may not affect the bactericidal action of the regimens. PMID- 8406639 TI - Serum immunoglobulin E response in sputum positive patients with pulmonary tuberculosis. AB - A marked elevation of serum IgE level (6939 +/- 5651 IU/ml) was observed in 20 untreated and drug sensitive patients with sputum AFB positive pulmonary tuberculosis. Of them 18 were freshly diagnosed and were totally untreated. Their mean serum IgE level was 6715 +/- 5545 IU/ml. The remaining two patients presented with a relapse of the disease but were sensitive to all anti tuberculosis drugs. Their serum IgE levels before retreatment were 2828 and 15079 IU/ml respectively. In the 13 chronic drug resistant patients who never had sputum conversion, the serum IgE level (1116 +/- 1897 IU/ml) was significantly lower (P < 0.01). It is suggested that the high serum IgE response in freshly diagnosed untreated and drug sensitive patients with illness of short duration may be related to strong early tuberculin hypersensitivity reaction and protective immunity, whereas low serum IgE response in chronic drug resistant cases with high bacterial load for prolonged duration may be related to the generation of suppressor CD8 lymphocytes eventually leading to suppression of B cells committed to the synthesis of protective serum IgE. PMID- 8406640 TI - Current status of drug resistance & phage types of Salmonella typhi in India. AB - A total of 11391 strains of S. typhi were received at the National Salmonella Phage Typing Centre at New Delhi during January 1990- August 1992, from various regions of India. Of these, 39.7 per cent were from north India, 37.5 per cent from central India and 22.8 per cent from south India. 64.5 per cent of S. typhi were found to be multidrug resistant (MDR), maximum resistance being observed in 1991 (71.6%) while it was least in 1990 (50.05%). There was a slight decline in the percentage of MDR strains in 1992. Region-wise analysis of the resistant strains showed maximum number in central India (71.32%), whereas it was least in the south (55.2%); 62.34 per cent of strains received from north India were MDR. Irrespective of the year or region, the predominant resistance pattern remained AMCSXTTE. The commonest phage type was E1, followed by 0 and A. This pattern was also seen in the MDR S. typhi. Certain degraded Vi strains, untypable Vi strains and Vi negative strains were also multidrug-resistant. An important change observed in this study was that a small number of strains belonging to phage types C1, K1, 28, 40, 41 and 42 which were always sensitive earlier, had developed multidrug resistance. Small outbreaks due to one phage type (e.g., 28 and 51) were short lived and subsided spontaneously. More diversity of phage types was observed in north India as compared to central and south India. PMID- 8406641 TI - Transferable colicinogeny in Salmonella bareilly. AB - Twenty eight (5.6%) of the 500 isolates of S. bareilly studied were found to be colicin producers. Of these 27 (96.4%) were Col V producers. None of these produced aerobactin and 18 (64.3%) were multidrug resistant. Among 23 (85.2%) strains both drug resistance as well as colicinogeny could be transferred by conjugation. This is the first time that the transferable colicinogeny has been demonstrated in S. bareilly. PMID- 8406642 TI - Numerical taxonomy of vibrio & allied organisms. AB - Forty four strains of Vibrio, Pasteurella, Yersinia, Aeromonas, Escherichia, Klebsiella and Enterobacter were tested for 44 characters and the overall results scored by the procedure of numerical taxonomy. The strains were clustered by the weighted pair group method with the arithmatic averaging. The V. cholerae 01 and the non-01 V. cholerae being interlinked at a very high level (86% to 97.5%) within a single cluster were considered under the same species. V.cholerae having Classic, E1 Tor and Gindha (Pfeiffer 1896, as per priority) biotypes respectively. The biotypes of V.parahaemolyticus remained clearly delineated from Vibrio and Aeromonas, and, belonged to a new separate genus. The V. parahaemolyticus biotypes were located more adjacent to Yersinia and Pasteurella than the V. cholerae cluster. V.fluvialis 1 appears as a biotype of V.parahaemolyticus, and, not a new species. PMID- 8406643 TI - An outbreak of acute diarrhoeal disease amongst tribal population in Tripura. AB - A massive outbreak of acute diarrhoeal diseases occurred during March-April, 1992 in the north district of Tripura. Investigation of the outbreak revealed Vibrio cholerae 01 biotype ElT or as the main etiologic agent in 50 per cent of patients. The outbreak which started amongst the tribal population might have spread due to prevailing illiteracy, poverty, low personal and domestic hygiene and vulnerable water sources (chhara water). PMID- 8406644 TI - Evaluation of amphotericin B as a first line drug in comparison to sodium stibogluconate in the treatment of fresh cases of kala-azar. AB - A total of 150 patients of kala-azar matched for age and sex and parasitologically proved were randomly allocated to two equal treatment groups. Patients in one group received amphotericin B(AMB) in a dose of 1 mg/kg body weight (BW) on alternate days starting with 0.05 mg/kg/bw on first day with daily increments, till a total dose of 20 mg/kg/bw was given; the patients in the second group received sodium stibogluconate (SAG) in the dose of 20 mg/kg/bw, im daily for 30 days. The efficacy, safety and cost-effectiveness of the two drugs were compared. Apparent cure (afebrile at the end of therapy) in 75 (100%) and 69 (92%) patients and ultimate cure (no relapse in six months of follow up) in 75 (100%) and 60 (80%) patients occurred in the AMB and SAG groups respectively. The difference between the ultimate cure in the two groups was significant (P < 0.001). Six (8%) and 9(12%) patients of SAG group showed primary (with no response to SAG during treatment) and secondary unresponsiveness (with no response to SAG after relapse) respectively and they were cured with amphotericin B.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406645 TI - Curative efficacy of norfloxacin in falciparum malaria. AB - Fifteen patients of uncomplicated falciparum malaria from Delhi were treated with norfloxacin (10 with 400 mg, 5 with 800 mg, both twice daily) for 3 days and the response was measured according to the WHO extended in vivo test criteria. The lower dose produced S response in two, RII response in five and RIII response in three patients, while the higher dose produced S response in four and RI response in one patient. In patients with S or RI response, the parasite clearance time was 68.6 +/- 9.1 h the defervescence time being 48 h. Thus, norfloxacin did reveal in vivo activity in falciparum malaria, but a dose of 400 mg twice daily proved to be curative only in a small percentage of cases and not consistently. Nausea and bitter taste were the only side effects noted in two patients. PMID- 8406646 TI - Isoenzyme pattern of clones of Entamoeba histolytica. AB - The isoenzyme pattern of clones of E. histolytica isolated from symptomatic and asymptomatic patients of amoebiasis were determined by starch gel electrophoresis. Ten parent cultures (uncloned), 5 from symptomatic and 5 from asymptomatic patients were isolated and established in Robinsons medium as xenic cultures. A total of 100 clones were isolated from parent cultures but only 65 could be established. Starch gel electrophoresis was carried out on the uncloned and cloned cultures. The analysis showed that the 5 uncloned culture from symptomatic patients had 4 isolates with pathogenic zymodeme and 1 isolate with non-pathogenic zymodeme pattern. The presence of beta band and absence of alpha band for phosphoglucomutase (PGM) and a fast moving Hexokinase (HK) band were taken as markers of pathogenicity. The 5 cloned cultures from asymptomatic patients had three isolates with pathogenic and 2 with non-pathogenic pattern. The cloned isolates from symptomatic patients (35) had 20 pathogenic zymodemes and asymptomatic patients (30) had 15 pathogenic zymodemes. Non-pathogenic zymodeme obtained in the present study were I, IV, V, X and XIII and the pathogenic zymodemes obtained were II and XIV. The data of the study indicated that (i) asymptomatic patients also harbour pathogenic zymodemes; and (ii) zymodeme II also coexsist with zymodeme XIV in the Indian population. PMID- 8406647 TI - Development of indigenous ELISA for rotavirus diagnosis & its comparison with commercial kit. AB - Preparation of reagents for the diagnosis of rotavirus by enzyme linked immunosorbent assay (ELISA) was carried out at the National Institute of Virology (NIV), Pune. This is a double antibody sandwich ELISA test. The coating antibody was raised in guinea pig against SA-11 (Simian rotavirus), and is used at 1:20,000 dilution. The indicator antibody was also raised against SA-11 in rabbits. The rabbit preimmune serum is used as negative control serum. Both, pre- and post-immune rabbit antisera are used at 1:10,000 dilution in the test. Goat IgG-HRP conjugate against rabbit IgG was procured commercially (Sigma, USA). Results of ELISA test can be read visually. A total of 63 pretested faecal specimens and 38 specimens of rotavirus in different concentrations were compared simultaneously by the NIV ELISA and Dakopatts ELISA. Of the 63 specimens, 36 were positive in NIV ELISA and 33 positive by Dakopatts ELISA. Human and animal rotavirus strains also showed higher titers in NIV ELISA than Dakopatts ELISA. From our data we conclude that NIV ELISA is 100 per cent specific and more sensitive than Dakopatts ELISA test. It is much more economical than any other commercial kit available for the diagnosis of rotavirus. Since the reagents are in lyophilized form, they are suitable for use in developing countries. PMID- 8406648 TI - Multiple drug resistant typhoid fever outbreak in Kashmir Valley. AB - We report the first ever report of multiple drug resistant salmonella typhi epidemic from Asia. The outbreak started in April 1988 and virtually ended by June 1988. The outbreak occurred in Baramullah town of Kashmir valley. In an epidemiological survey 230 cases were diagnosed as enteric fever. 46 cases (20%) were culture positive. A very rare multiple drug resistant strain of S. typhi was identified as phage type biotype II untypeable (UVS2). The strain was sensitive to Norfloxacin. This outbreak of S. typhi has many similarities with Mexico epidemic. Ingestion of contaminated water seems to be the possible cause for the outbreak. PMID- 8406649 TI - Cotard's syndrome in migraine (a case report). PMID- 8406650 TI - Iron deficiency and its implications. PMID- 8406651 TI - In vivo hypothalamic release and synthesis of catecholamines in spontaneously hypertensive rats. AB - Juvenile spontaneously hypertensive rats (SHR) have higher plasma levels of catechols and markedly larger catechol responses to yohimbine than do normotensive Wistar-Kyoto rats, indicating increased sympathoadrenal outflow and increased alpha 2-adrenergic receptor-mediated restraint of peripheral catecholamine release during hypertension development in SHR. Yohimbine-induced catecholamine release and metabolism in the posterolateral hypothalamus of the brain were assessed in juvenile (6 to 7 weeks) and adult (15 to 16 weeks) SHR and Wistar-Kyoto rats. In vivo microdialysis was used to obtain samples for measurements of norepinephrine, dihydroxyphenylglycol, methoxyhydroxyphenylglycol, and dihydroxyphenylacetic acid in conscious animals before and after yohimbine injection (1 mg/kg IV) beginning 24 hours after probe implantation. Catecholamine synthesis was examined from elevations of 3,4 dihydroxyphenylalanine levels after probe perfusion with NSD-1015, an inhibitor of L-aromatic acid decarboxylase. In adults, SHR had higher dialysate norepinephrine (277 +/- 38 versus 181 +/- 35 pg/mL), dihydroxyphenylglycol (3260 +/- 509 versus 2231 +/- 201 pg/mL), methoxyhydroxyphenylglycol (2659 +/- 369 versus 1890 +/- 144 pg/mL), and dihydroxyphenylacetic acid (46,312 +/- 5512 versus 13,187 +/- 1963 pg/mL) levels and markedly larger increases in 3,4 dihydroxyphenylalanine levels after NSD-1015 than Wistar-Kyoto rats. In juveniles, SHR had larger proportionate increments in microdialysate norepinephrine levels after yohimbine than Wistar-Kyoto rats (85% versus 25%). Although juvenile SHR and Wistar-Kyoto rats had similar NSD-1015-elicited increments in 3,4-dihydroxyphenylalanine levels, systemic yohimbine enhanced the NSD-1015-elicited 3,4-dihydroxyphenylalanine elevations in juvenile SHR but not in Wistar-Kyoto rats. These findings suggest augmented norepinephrine release and catecholamine synthesis in the posterolateral hypothalamus of adult SHR and augmented alpha 2-adrenergic receptor restraint of both norepinephrine release and catecholamine synthesis in juvenile SHR. PMID- 8406652 TI - Cardiovascular reactivity to stress predicts future blood pressure status. AB - Cardiovascular reactivity to stress may have a pathophysiological role in neurogenic hypertension. We studied the value of measuring blood pressure change during standardized mental and physical challenges to prediction of resting blood pressure status 6.5 years later among 206 middle-aged adults and their 164 children, with the latter group originally being tested while enrolled in elementary through high school. After adjustment for age, resting blood pressure, and body mass index at study entry, as well as length of follow-up, larger systolic and diastolic blood pressure responses to a combination of mental and physical challenges were associated with higher subsequent resting diastolic blood pressure 6.5 years later among adults. Among boys, but not among girls, larger systolic and diastolic blood pressure responses to challenge were associated with higher subsequent resting blood pressure. These data suggest that people who are at high risk for elevated blood pressure might have an exaggerated stress-induced cardiovascular response at a younger age. PMID- 8406653 TI - In vitro perfusion studies of resistance artery function in genetic hypertension. AB - To examine the function of resistance-sized arteries in hypertension under in vitro conditions that approximate in vivo conditions as much as possible, we mounted segments of second-order mesenteric resistance arteries from spontaneously hypertensive rats (SHR) and Wistar-Kyoto normotensive control rats aged 12 to 13 weeks in a perfusion myograph and exposed them to conditions of constant flow and pressure. The endothelial integrity was validated both functionally and histologically. Vascular sensitivity to norepinephrine was examined when the hormone was applied either intraluminally or extraluminally and before and after removal of the endothelium. Both endothelium-dependent and independent dilatation was assessed by the intraluminal application of acetylcholine and sodium nitroprusside, respectively. Sodium nitroprusside was applied to arteries after endothelium removal. Arterial responses were measured by changes in intraluminal diameter recorded with a video camera and imaging system. Vessels from SHR demonstrated depressed endothelium-dependent relaxation but similar endothelium-independent relaxation and greater sensitivity to norepinephrine with both intraluminal and extraluminal application. Removal of the endothelium abolished the differences in sensitivity to norepinephrine between the two strains. The results demonstrate that resistance arteries from SHR when examined under in vitro perfusion display enhanced sensitivity to norepinephrine due to depressed endothelium-dependent dilatation, and the data suggest that functional modifications in the endothelium may play an important role in hypertensive vascular disease. PMID- 8406654 TI - Nuclear angiotensin receptors induce transcription of renin and angiotensinogen mRNA. AB - The observation that nuclei from hepatic tissue exhibit specific angiotensin II (Ang II) binding led us to explore whether Ang II modulates mRNA in general, mRNA specific for renin system components, or both. Nuclei from hepatic tissue exhibited a single high-affinity (Kd = 0.4 nmol/L) Ang II-specific binding site, which was associated with increased RNA transcription. Whereas total RNA extracted from nuclei increased 1.5-fold in response to Ang II (10(-9) mol/L), specific mRNA for renin and angiotensinogen increased 7.8- and 2.5-fold, respectively. Ang II binding and induced transcription showed parallel Ang II dose responses that were both inhibited by 10(-5) mol/L DuP 753 or saralasin. Maximum Ang II binding and RNA transcription occurred at the same Ang II concentration (10(-9) mol/L). Higher doses of Ang II resulted in a progressive decrease in RNA transcription. Together, these results demonstrate that hepatic nuclei have functional Ang II-specific receptors. It is concluded that Ang II may elicit responses at nuclear receptors, which heretofore were associated only with Ang II receptors located on plasma membranes. However, the individual contribution of plasma and nuclear membrane Ang II receptors to the overall cellular Ang II transcriptional response and their possible interactions remain to be determined. PMID- 8406655 TI - Feasibility and efficacy of sodium reduction in the Trials of Hypertension Prevention, phase I. Trials of Hypertension Prevention Collaborative Research Group. AB - Phase I of the Trials of Hypertension Prevention was a multicenter, randomized trial of the feasibility and efficacy of seven nonpharmacologic interventions, including sodium reduction, in lowering blood pressure in 30- to 54-year-old individuals with a diastolic blood pressure of 80 to 89 mm Hg. Six centers tested an intervention designed to reduce dietary sodium to 80 mmol (1800 mg)/24 h with a total of 327 active intervention and 417 control subjects. The intervention consisted of eight group and two one-to-one meetings during the first 3 months, followed by less-intensive counseling and support for the duration of the study. The mean net decrease in sodium excretion was 43.9 mmol/24 h at 18 months. Women had lower sodium intake at baseline and were therefore more likely to decrease to less than 80 mmol/24 h. Black subjects were less likely to decrease to less than 80 mmol/d, independent of sex or baseline sodium excretion. The mean (95% confidence interval) net decrease associated with treatment was -2.1 (-3.3, -0.8) mm Hg for systolic blood pressure and -1.2 (-2.0, -0.3) mm Hg for diastolic blood pressure at 18 months (both P < .01). Multivariate analyses indicated a larger systolic blood pressure effect in women (-4.44 versus -1.23 mm Hg in men), adjusted for age, race, baseline blood pressure, and baseline 24-hour urinary sodium excretion (P = .02). Dose-response analyses indicated an adjusted decrease of -1.4 mm Hg for systolic blood pressure and -0.9 mm Hg for diastolic blood pressure for a decrease of 100 mmol/24 h in 18-month sodium excretion. These results support the utility of sodium reduction as a population strategy for hypertension prevention and raise questions about possible differences in dose response associated with gender and initial level of sodium intake. PMID- 8406656 TI - Nephrectomy, converting enzyme inhibition, and angiotensin peptides. AB - To determine the contribution of kidney-derived renin and angiotensin converting enzyme to circulating and tissue levels of angiotensin peptides, we measured angiotensin (Ang)-(1-7), Ang II, Ang-(1-9), and Ang I in plasma, kidney, lung, heart, aorta, brown adipose tissue, adrenal, pituitary, and brain of five groups of male Sprague-Dawley rats: control rats, rats given the converting enzyme inhibitor ramipril (10 mg/kg), rats nephrectomized 24 hours, rats nephrectomized 48 hours, and rats nephrectomized 48 hours and given ramipril. Plasma and tissues, apart from adrenal, showed a 63% to 98% reduction in Ang II, the ratio of Ang II to Ang I, or both after ramipril administration, indicating a major role for converting enzyme in Ang II formation. Nephrectomy caused a more than 95% decrease in plasma renin levels and a fourfold to eightfold increase in plasma angiotensinogen levels. Apart from plasma and brain, tissues showed a 59% to 78% decrease in Ang II levels after nephrectomy, indicating a major role for kidney-derived renin in Ang II formation. The persistence of Ang II in plasma and tissues of anephric rats indicates that Ang II may be formed by a process independent of kidney-derived renin; this process may be amplified by the increased plasma angiotensinogen levels that accompany nephrectomy. For lung, adrenal, and aorta, Ang II levels showed a further decrease when nephrectomized rats were given ramipril. However, for plasma and the other tissues, ramipril produced little or no decrease in Ang II levels of anephric rats, suggesting that Ang II may be formed by a pathway independent of converting enzyme. Such a pathway may involve the direct formation of Ang II from angiotensinogen by a non renin-like enzyme. PMID- 8406657 TI - Histology of subcutaneous small arteries from patients with essential hypertension. AB - The purpose of the present study was to determine the cellular basis for the increased ratio of media thickness to lumen diameter (media-lumen ratio) consistently found in the peripheral resistance arteries from patients with essential hypertension using an unbiased stereological principle (the "disector"). Segments of subcutaneous resistance arteries (approximately 200 microns internal diameter) were isolated from gluteal biopsies of skin and subcutaneous fat taken from 16 untreated patients with essential hypertension and 16 age- and sex-matched normotensive control subjects. Measured under standardized conditions (ie, relaxed and under controlled mechanical conditions) on an isometric myograph, vessels from hypertensive patients had a significant (P < .05) reduction in lumen diameter and an increase in media-lumen ratio (P < .05) compared with vessels from normotensive control subjects. These changes were not associated with alterations in the estimated media volume per segment length. After these measurements had been made, the arteries were fixed, serial sectioned, and stained. The volume fraction of smooth muscle cells within the media was estimated by point counting on photomicrographs of the vessels. Using the disector principle, we determined the numerical density (number per unit volume) of smooth muscle cells within the media of each vessel and calculated the average smooth muscle cell volume (1775 +/- 122 [mean +/- SEM] and 1532 +/- 112 microns 3, hypertensive and normotensive, respectively, P > .05) on the basis of these measurements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406658 TI - Renal responses to intra-arterial administration of nitric oxide donor in dogs. AB - Inhibition of nitric oxide synthesis by intra-arterial administration of nitro-L arginine (NLA) leads to attenuation of the slope of the relation between renal arterial pressure (RAP) and sodium excretion without an alteration in renal autoregulatory efficiency. In the present study, we examined whether only the presence of nitric oxide or, alternatively, changes in nitric oxide production during changes in RAP are required for pressure natriuresis to occur. Anesthetized sodium-replete dogs (n = 8) were treated with NLA (50 micrograms.kg 1 x min-1) to inhibit endogenous nitric oxide formation, and S-nitroso-n acetylpenicillamine (SNAP) was infused intra-arterially at a constant rate (2 micrograms.kg-1 x min-1) to replenish intrarenal nitric oxide levels. Renal responses to reductions in RAP within the autoregulatory range were assessed before and during NLA infusion followed by SNAP+NLA infusion. As reported previously, NLA infusion alone increased renal vascular resistance and decreased renal blood flow, urine flow, sodium excretion, and fractional excretion of sodium, with no change in glomerular filtration rate. Autoregulatory efficiency remained intact, whereas the pressure-induced natriuretic responses were attenuated. During SNAP+NLA infusion, renal blood flow increased from 2.8 +/- 0.3 to 3.5 +/- 0.3 mL.min-1 x g-1 (P < .001), without significant changes in glomerular filtration rate (0.75 +/- 0.07 to 0.81 +/- 0.05 mL.min-1 x g-1); the autoregulatory efficiency of renal blood flow and glomerular filtration rate remained intact. SNAP increased urine flow (4.8 +/- 1.8 to 10.0 +/- 2.5 microL.min-1 x g-1), sodium excretion (0.63 +/- 0.26 to 1.70 +/- 0.37 mumol.min-1 x g-1), and fractional excretion of sodium (0.55 +/- 0.20% to 1.38 +/- 0.27%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406659 TI - Developmental sensitivity to high dietary sodium chloride in borderline hypertensive rats. AB - The present study compared the postweaning blood pressures and body weights of borderline hypertensive rats exposed to a high (8%) sodium chloride maternal diet either from conception to weaning or only during the weaning period with borderline hypertensive rats consistently exposed to a normal (1%) sodium chloride maternal diet. Because the effects of early sodium chloride exposure may be most evident during a subsequent challenge, rats from each group were assigned to receive either an 8% sodium chloride or a 1% sodium chloride diet from 8 to 17 weeks of age. Exposure to an 8% sodium chloride diet from conception through weaning increased the adult blood pressure of borderline hypertensive rats compared with that of controls exposed to a 1% sodium chloride diet; exposure to an 8% sodium chloride diet only during weaning did not increase blood pressure. An 8% sodium chloride diet beginning at 8 weeks of age increased systolic blood pressure. The effects of perinatal and adult exposure to high dietary sodium chloride were additive. Behavioral observations and urinary electrolyte measures confirmed that pups exposed to an 8% sodium chloride diet during weaning ingested the high-sodium chloride diet. The blood pressure and heart rate response to autonomic nervous system ganglionic blockade were assessed at 17 weeks of age. Borderline hypertensive rats exposed to an 8% sodium chloride diet from conception through weaning showed an increased bradycardic response, but no difference in depressor response, to ganglionic blockade. These data suggest that the window of developmental sensitivity for modulation of blood pressure regulation by high dietary sodium chloride occurs during prenatal and early postnatal development. PMID- 8406660 TI - Blood pressure change and survival after age 75. AB - Higher diastolic pressure predicted better survival in men 75 years or older in two prior analyses in the Rancho Bernardo population. Diastolic change was implicated as a possible explanation. We studied this by assessing survival according to blood pressure change in 795 men and women aged 75 years and older at the time of a second measurement taken an average of 11 years after the first, who were then followed for 5 years. Sex-specific analyses compared participants with a diastolic decrease of 5 mm Hg or greater and participants with a systolic decrease of 10 mm Hg or greater with those whose blood pressure levels did not change or increased. In men, after adjustment for baseline pressure, a decrease in diastolic pressure of 5 mm Hg or greater was associated with higher all-cause mortality (relative risk, 2.33; 95% confidence interval, 1.39 to 3.91) and cardiovascular mortality (3.13, 1.47 to 6.66). The mortality risk was strongest in men who took antihypertensive medication and had a fall in diastolic pressure (12.33, 2.73 to 55.72) compared with treated men whose pressures did not decrease. Among men with isolated systolic hypertension, those treated whose diastolic pressure remained stable had the best survival. A systolic fall in men and a decrease in either diastolic or systolic in women was not associated with poorer survival after adjustment for baseline pressure. We conclude that a fall in diastolic pressure of 5 mm Hg was associated with poor survival in men after age 75. This risk was strongest in men who took antihypertensive medication. PMID- 8406661 TI - Sodium-lithium countertransport genotype and the probability of hypertension in adults. AB - The objective of the present study was to determine whether information about a biometrically inferred single gene with large effects on erythrocyte sodium lithium countertransport is useful in predicting the probability of having hypertension. We used multivariate logistic regression to model the relationship between the probability of having hypertension and predictor traits in a sample of 382 unrelated adult women and 347 unrelated adult men from Rochester, Minn. First, we identified a set of demographic, biochemical, and physiological predictors. Second, we analyzed whether the relationship between the probability of having hypertension and the identified predictor traits was heterogeneous between the biometrically inferred single locus genotypes with large effects on sodium-lithium countertransport level. Third, if there was no heterogeneity, we assessed whether sodium-lithium countertransport genotypes made an additional contribution to predicting the probability of having hypertension after other predictors were considered. In women, the predictors of the probability of having hypertension were age, plasma apolipoprotein CIII, body mass index, and an interaction term involving age and body mass index. The relationship between the probability of having hypertension and the identified predictors was not heterogeneous between sodium-lithium countertransport genotypes, and genotype did not contribute to the prediction of the probability of having hypertension after the identified predictors were considered. In men, predictors of the probability of having hypertension were age, plasma levels of high-density lipoprotein cholesterol, apolipoproteins AI and CII, sodium-lithium countertransport level, and sodium-lithium countertransport genotype. The relationship between the probability of having hypertension and sodium-lithium countertransport level and age were heterogeneous between biometrically inferred sodium-lithium countertransport genotypes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406662 TI - Effects of calcium channel blockade on the aortic intima in spontaneously hypertensive rats. AB - Hypertension is associated with an intimal dysfunction characterized by endothelium-dependent constriction to serotonin, decreased endothelium-dependent relaxation to acetylcholine, and a subendothelial infiltration of monocyte macrophages. The goal of our study was to evaluate the effect of long-term calcium channel blockade with Ro 40-5967, a new long-acting calcium channel blocker, on these alterations in aortas of spontaneously hypertensive rats (SHR). Arterial blood pressure was decreased by Ro 40-5967. In aortas from Ro 40-5967 treated SHR, the serotonin ratio (maximal contraction to serotonin on rings with endothelium over maximal contraction on paired rings without endothelium) was reduced (1.14 +/- 0.10) compared with control SHR (1.72 +/- 0.12, P < .01) because of inhibition of maximal contraction in rings with endothelium. This effect of Ro 40-5967 was partially reversed by an inhibitor of nitric oxide (NO) synthase, NG-nitro-L-arginine-methyl ester, and partially inhibited in the presence of the thromboxane/prostaglandin H2 receptor antagonist AH 23848. Maximal relaxation to acetylcholine in rings with endothelium was increased by Ro 40-5967. In rings without endothelium, Ro 40-5967 treatment enhanced the sensitivity to sodium nitroprusside-induced relaxation. Cyclic GMP content, an indicator of NO release, was not increased in aortas from Ro 40-5967-treated SHR. Thus, improvement of endothelial function was probably achieved by facilitating the action of NO at the level of the smooth muscle cells and by reducing prostaglandin H2-induced constriction. Finally, the number of monocyte macrophages in the subendothelium was decreased by Ro 40-5967. 40-5967.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406663 TI - Purinergic endothelium-dependent and -independent contractions in rat aorta. AB - The role of endothelium-derived contracting factor or factors in modulating relaxations and contractions to adenine nucleotides was examined in aortas from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) and Wistar rats. During contractions to phenylephrine, the relaxations to ATP were impaired significantly in SHR compared with WKY aortas with endothelium. In rings treated with NG-nitro-L-arginine (to inhibit nitric oxide synthase), the endothelium significantly augmented contractions evoked by ATP; this enhancement was greater in SHR compared with WKY aortas. Indomethacin (inhibitor of cyclooxygenase) and SQ 29,458 (antagonist of thromboxane/prostaglandin endoperoxide receptors) but not dazoxiben (inhibitor of thromboxane synthase) significantly augmented the maximal relaxation in WKY rats, abolished the impairment of the relaxation in SHR, and prevented the potentiation by the endothelium of the contractions evoked by ATP. In older animals (10 to 12 months old), the endothelium-dependent concentration-relaxation curves to ATP in SHR and WKY aortas treated with indomethacin were superimposable, as were the concentration-contraction curves (with NG-nitro-L-arginine present). Endothelium dependent concentration-relaxation and -contraction curves to ADP obtained in these preparations overlapped also. In Wistar rats, the magnitude of the endothelium-dependent relaxations to either ATP or ADP were significantly smaller compared with the other strains, and the endothelium-dependent contractions were even smaller. Results show that adenine nucleotides stimulate the production of both endothelium-derived relaxing and contracting factors. Although there is no obvious age-related alteration in the capacity of aortas to release endothelium derived relaxing factor, aging enhances endothelium-derived contracting factor activity in WKY rats. PMID- 8406664 TI - Insulin does not reduce forearm alpha-vasoreactivity in obese hypertensive or lean normotensive men. AB - Evidence supports the hypothesis that an impaired capacity of insulin to antagonize norepinephrine-induced vasoconstriction increases alpha-adrenergic tone in overweight young men with insulin resistance and mild hypertension. Therefore, the effects of regionally infused insulin at 100 microU/mL on forearm blood flow (milliliters per deciliter per minute) and responses to norepinephrine were measured in seven obese hypertensive and eight lean normotensive men younger than 45 years old. The obese hypertensive men were hyperinsulinemic and insulin resistant compared with the normotensive men, as evidenced by abnormal values for fasting insulin (15.5 +/- 1.6 versus 7.2 +/- 0.8 microU/mL, P < .001), the insulin area under the curve in response to a 2-hour oral glucose tolerance test (12.0 +/- 1.5 versus 6.7 +/- 1.1 mU x min/dL, P < .01), and the disappearance rate of glucose during a 15-minute insulin tolerance test (2.7 +/- 0.3 versus 4.1 +/- 0.3 mg%/min, P < .05). The logarithm of the norepinephrine EC50 was not significantly different in obese hypertensive men (mean, 95% confidence interval: -8.15, -8.42 to -7.87) versus lean normotensive men (-7.91, -8.23 to -7.59). The 2-hour regional insulin infusion at 100 microU/mL did not significantly alter the EC50 for norepinephrine in either group. Insulin at this concentration induced significant and similar increases of forearm blood flow in the hypertensive and normotensive groups (1.7 +/- 0.4 versus 1.7 +/- 0.6 mL/100 mL per minute, P = NS). At approximately 100 microU/mL, insulin does not antagonize norepinephrine induced vasoconstriction in the forearm circulation of either obese hypertensive or lean normotensive men.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406665 TI - Bias and variability in blood pressure measurement with ambulatory recorders. AB - This study sought to determine whether patient characteristics such as age, sex, blood pressure, and pulse pressure differently affect the accuracy of an oscillometric (SpaceLabs 90207) and a microphonic (TM2420 version 7) blood pressure monitor. Blood pressure recorded by two oscillometric and two microphonic ambulatory monitors was compared with simultaneous readings by two pairs of trained, blinded observers using random-zero sphygmomanometry. One hundred and eighteen subjects (53 men and 65 women, aged 17 to 94 years; systolic pressure, 89 to 211 mm Hg; diastolic, 44 to 116 mm Hg) were studied. There were no significant differences within each observer pair or between the two observer pairs as well as no correlation between interobserver differences and patient characteristics. The differences between the monitor and trained observers' readings were 2.8 +/- 9.9 mm Hg systolic and 3.9 +/- 6.8 mm Hg diastolic for the SpaceLabs and 5.0 +/- 5.2 mm Hg systolic and 3.4 +/- 6.1 mm Hg diastolic for the TM2420. Patient characteristics that predicted measurement error were defined by multiple regression. For oscillometry, systolic measurement error was highly correlated with systolic pressure, pulse pressure, and subject age. The diastolic error was significantly correlated with pulse pressure, diastolic pressure, and subject sex. For the oscillometric monitor, patient characteristics accounted for 36.6% of the variation of the systolic error and 34.7% of the variation of the diastolic error. For the microphonic monitor, only age correlated with diastolic error, and no significant correlations were seen with systolic error. Patient characteristics accounted for only 1.2% of the systolic and 8.9% of the diastolic error.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406666 TI - Major approaches for generating and analyzing transgenic mice. An overview. AB - Over the past decade, the development of gene-transfer technology in whole animals has afforded unprecedented opportunities for investigators to probe complex regulatory systems in vivo. Important advances in our understanding of the mechanisms of gene expression and regulation and the development of animal models of human diseases are but two examples of how this technology has affected medical science. Transgenic animals are defined as animals in which a segment of DNA has been physically integrated into the genome of all cells, including the germ line, so that it can be transmitted to offspring as a simple Mendelian trait. The DNA segment generally consists of a whole cloned gene, cDNA, or a novel gene modified by recombinant DNA methodologies. Whole genomic clones of genes are often used to study tissue- and cell-specific expression and regulation or can be used to overexpress a gene product. Alternatively, the coding region of one gene can be fused to the transcriptional regulatory region of another gene, causing it to be expressed in a new spectrum of tissues and cell types. A number of methods can be used to introduce the DNA segment, including direct microinjection of one-cell fertilized embryos, retroviral-mediated transfer, or gene transfer in embryonic stem cells. The technique most often used to generate transgenic animals and perform "gene addition" experiments is direct microinjection. Alternatively, gene deletions or "knockouts" are performed by gene transfer in embryonic stem cells by specifically targeting the site of integration in the genome.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406667 TI - Targeting gene expression to specific cardiovascular cell types in transgenic mice. AB - Transgenic techniques, which allow the introduction of exogenous genes into the genome of experimental animals, promise to bridge the gap between the in vitro observations made by molecular and cellular biologists on cardiac and vascular cells in tissue culture and the physiology and pathology of the whole organ system. One such application of these techniques is tissue targeting: by genetic manipulation to direct expression of a protein--such as a signaling peptide, a growth factor receptor, or an oncogene involved in cell growth--to a tissue where it normally would not be expressed (or where expression is tightly controlled) by fusing it to the transcriptional control sequences of another gene normally expressed in that tissue. In the cardiovascular system, regulatory sequences for cardiomyocyte-specific proteins, vascular endothelium-specific proteins, and smooth muscle-specific proteins can be used to target heterologous genes to their respective tissues in transgenic animals. The effects that such perturbations have on organ physiology and intracellular and intercellular communication can be observed by applying established physiological and molecular approaches. In this review, we highlight some tissue-specific genes from cardiac and vascular cell types whose regulatory sequences may be used to target heterologous proteins; we discuss neutral "reporter" proteins and signal transduction components as paradigms for the application of this technique; and we briefly touch on the potentials and pitfalls of transgenic approaches to molecular physiology. PMID- 8406668 TI - Embryonic stem cell model systems for vascular morphogenesis and cardiac disorders. AB - To better understand the formation of the cardiovascular system and its disease states, models amenable to manipulation must be developed. In this article we present two models. One is a small animal model for an inflammatory disorder that can lead to heart failure. Production of this model is based on the ability of blastocyst-derived embryonic stem cells, which can be genetically altered in vitro by a technique called gene targeting, to reconstitute an entire animal when reintroduced into a blastocyst and allowed to colonize the germ line of the resulting chimeric embryo. The other model is based on the capacity of embryonic stem cells to differentiate in culture into embryo-like structures called embryoid bodies. Embryoid bodies contain angioblasts, or prevascular endothelial cells, which can be induced to undergo aspects of vascular development by manipulation of culture conditions. PMID- 8406669 TI - Transgenesis in nonmurine species. AB - Although the mouse remains the species of choice for most transgenic experimentation, it may be preferable or even necessary to use alternative species for certain applications. We review the strategies by which transgenic technology has been applied to other animals, specifically, the rat, rabbit, pig, sheep, goat, and cow. Additionally, we outline the potential applications of alternative transgenic species with reference to the field of hypertension and cardiovascular research. PMID- 8406670 TI - Atrial natriuretic factor and transgenic mice. AB - Atrial natriuretic factor (ANF) is a peptide hormone that induces potent but transient hypotensive and natriuretic responses on short-term administration. The role of the hormone in long-term cardiovascular regulation has remained elusive in part because of the temporal limitations of long-term infusion models and the extremely short half-life of the molecule in vivo. To circumvent these temporal limitations, a transgenic mouse model was developed that exhibits lifelong elevated plasma ANF levels. These mice are chronically hypotensive, with arterial pressures averaging 20 to 30 mm Hg less than those observed in nontransgenic siblings. In contrast, no obvious natriuretic or diuretic phenotype was observed in transgenic animals housed in metabolic cages. Thus, the mice adequately compensate for the renal effects but not the hemodynamic effects of the hormone. The ANF transgenic mice provide a tractable model system with which to study the consequences of long-term alterations of ANF expression in vivo. PMID- 8406671 TI - Transgenic mouse models of vasopressin expression. AB - Arginine vasopressin is a nine-amino acid neuropeptide hormone important in the regulation of water metabolism. It also may have a role in other physiological functions, such as blood pressure regulation and the response to stress. Whole animal studies have provided a good understanding of vasopressin physiology and regulation of the normal vasopressin gene, and in vitro cell culture studies have demonstrated important features of the intracellular regulation of vasopressin gene expression. Transgenic mice provide useful models for the study of the in vivo regulation of gene expression. Previously reported mouse lines transgenic with vasopressin gene constructs have not expressed the transgene in a tissue distribution similar to that detected for the endogenous mouse vasopressin gene. An 8.2-kb genomic construct of the rat vasopressin gene, including 3 kb each of 5' and 3' flanking sequences, has been used to develop a line of transgenic mice. These animals express the transgene in a tissue-specific manner, demonstrate appropriate osmotic regulation of transgenic vasopressin mRNA, and have normal water metabolism. Animals homozygous for the 8.2-kb transgene have increased basal plasma levels of vasopressin peptide but have no apparent change in basal water metabolism. The findings with this and other previously reported mouse lines transgenic for vasopressin constructs provide a basis for developing future transgenic lines to study the in vivo regulation of the vasopressin gene. PMID- 8406672 TI - Double knockouts. Production of mutant cell lines in cardiovascular research. AB - Double knockouts by homologous recombination is a method for producing cell lines with an inactivating mutation in any desired gene. The biochemical analysis of genetically altered cell lines has been important in determining the function of specific proteins. Until recently, mutant cell lines have been produced by random mutagenesis and then selection for a particular phenotypic change. Recent technological advances in gene targeting by homologous recombination now enable the production of mutants in any desired gene. Diploid cells contain two copies or alleles of each gene encoded on an autosome (nonsex) chromosome. In most cases, both alleles must be inactivated to produce a phenotypic change in a mutant cell line, hence the term "double knockout." We and others have described the production of mutationally altered cell lines by inactivating both alleles by the production of two targeting vectors, two separate homologous recombination events, and selection. A simpler procedure, involving considerably less effort and time, has been used to inactivate several alpha-subunits of G proteins and other genes. This method facilitates the inactivation of more than one gene in a single cell line. PMID- 8406673 TI - Monoamine oxidase-B in astrocytes. AB - In the present report we describe the astrocytic localization and content of monoamine oxidase-B (MAO-B) by means of a 3H-L-deprenyl emulsion autoradiography in primary cultures of rat astrocytes, in cryosectioned astrocytoma surgical specimen, and in cryosections of human spinal cords from patients dying in amyotrophic lateral sclerosis (ALS) and controls. The occurrence of MAO-B enzyme protein depends on the degree of cellular differentiation as demonstrated by studies on astrocytes in primary cultures analyzed at two different stages of maturation. Highly differentiated cells exhibited high relative enzyme concentration whereas glioblasts lacked or showed very low contents of MAO-B enzyme. This was further substantiated by studies performed on human astrocytoma tissue using 3H-L-deprenyl emulsion autoradiography in combination with immunohistochemical detection of glial fibrillary acidic protein (GFAP). Regional increases of MAO-B concentration were found in ALS lumbar sections with quantitative 3H-L-deprenyl autoradiography. On the basis of results obtained from double staining for GFAP and MAO-B, the increase in MAO-B seemed to be due to an increased number of astrocytes as well as an increased content of MAO-B in reactive species of astrocytes. A cell culture model has been used that produces cells with morphology and GFAP-content similar to reactive cells. These astrocytes exhibited high relative content of the MAO-B enzyme protein. In the light of the presented data, taking into account the finding that a subpopulation of reactive cells contained low levels of MAO-B, a heterogeneity among reactive astrocytes was observed. PMID- 8406674 TI - Differential expression of carbonic anhydrase isozymes in microglial cell types. AB - Microglia cells have been shown to express carbonic anhydrase. Using carbonic anhydrase histochemistry and immunohistochemistry, different types of central nervous system microglial cells were detected, which expressed two main carbonic anhydrase (CA) isozymes during the early postnatal stage of development and after peripheral nerve injury in the spinal cord of adult rats. Amoeboid and reactive microglial cells were heavily immunostained for CA-II and CA-III and showed colocalization with complement receptor type 3 and Griffonia Simplicifolia B4 isolectin. Resting microglial cells in the brain and spinal cord showed faint CA III staining and were negative for CA-II. These results show that not only CA-II, but also CA-III isozyme is represented in the central nervous system and carbonic anhydrase activity may correlate with metabolic and immunological changes of microglial cells. These data also further strengthen the idea of the mesodermal origin of central nervous system macrophages. PMID- 8406675 TI - Extracellular matrix modulates the proliferation of rat astrocytes in serum-free culture. AB - The mechanism of glial proliferation in the developing nervous system, as well as in response to injury, inflammation, and tumor invasion, is unknown. Several growth factors and extracellular matrices have been shown to stimulate the proliferation of cultured cells of various origin, including astrocytes. We investigated the effect of extracellular matrix components, including fibronectin (FN), laminin (LN), and collagen types I and IV, on the growth of astrocytes during stimulation by various growth factors. When astrocytes were grown on FN- and LN-coated wells in a serum-free, chemically defined medium, their increase in number significantly exceeded that of cells grown on plastic wells. The addition of platelet-derived or basic fibroblast growth factor to cells cultured on FN- or LN-coated wells significantly potentiated astrocyte proliferation. The collagen preparations had no such effect. These observations indicate that FN and LN have a fundamental part in converting the quiescent astrocyte into the proliferating phenotype, which may be required for remodelling damaged brain tissues in vivo. PMID- 8406676 TI - Effects of interleukin-1 beta and tumor necrosis factor-alpha on the expression of glial fibrillary acidic protein and transferrin in cultured astrocytes. AB - Recent evidence suggests that interleukin (IL)-1 and tumor necrosis factor (TNF) may play a role in astrogliosis following injury to the CNS. The short-term biochemical effects of these immune-related cytokines were determined on cultured rat polygonal and process-bearing astrocytes. Both IL-1 and TNF stimulated the rate of thymidine incorporation in polygonal astrocytes up to 137% and 215%, respectively, over the level observed in untreated controls. By contrast, thymidine incorporation was relatively unaffected by these cytokines in process bearing astrocytes. The cytokines did not significantly affect the level of glial fibrillary acidic protein (GFAP) within polygonal astrocytes, even though they appeared to downregulate the expression of GFAP mRNA by as much as 62%. Both cytokines increased the intracellular expression of transferrin (Tf) within some polygonal astrocytes. In untreated control cultures, fewer than than 2% of polygonal astrocytes were immunoreactive for Tf. By contrast, approximately 30% of polygonal astrocytes treated with IL-1 or TNF-alpha became strongly immunoreactive for Tf. Neither IL-2 nor a number of other known growth factors appeared to alter the level of immunoreactive Tf in these cells. Process-bearing astrocytes were negative for Tf, regardless of the treatment used. Northern blot analysis demonstrated that the level of Tf mRNA in cultures of polygonal astrocytes increased 148% above the level observed in untreated controls following treatment with either IL-1 or TNF, whereas no change was observed following treatment with IL-2. These results suggest that increased levels of particular cytokines known to be present in injured CNS can produce pronounced biochemical alterations within a subtype of cultured astrocytes. PMID- 8406677 TI - Characterization of stretch-activated ion channels in cultured astrocytes. AB - The presence of a stretch-activated channel in rat cerebellar astrocytes in culture is described. This stretch-sensitive channel is K(+)-selective and its open probability increases with suction following a Boltzmann-like distribution with half activation at 45 mm Hg. Kinetic analysis of the single-channel data indicated that there are two open and two closed states and that the shortest time constants of both open and closed states are the most sensitive to suction. A symmetrical two-barrier-one-site permeation model can quantitatively describe the inward rectification of the single-channel current to voltage relations. It is suggested that this stretch-activated channel plays a role in the regulatory volume response of astrocytes to hyposmotic conditions. PMID- 8406678 TI - Changes in ultrastructure and voltage-dependent currents at the glia limitans of the frog optic nerve following retinal ablation. AB - The surface of the frog optic nerve consists of astrocytic processes separated by narrow extracellular clefts underlying a pial sheath of loose connective tissue. Macroscopic voltage dependent currents can be recorded from this surface using the loose patch-clamp technique. In this study the changes in ultrastructure and voltage dependent Na currents have been studied for up to 1 year following removal of the retina. During the first 1-4 weeks, many of the myelinated and unmyelinated axons of the retinal ganglion cells degenerate, and the debris is phagocytosed by macrophages and glial cells. However, some morphologically intact axons remain even 12 weeks after surgery. Finally, after 16 weeks all the axons have disappeared, leaving a nerve consisting only of glial cells, some of which contain phagosomes. At 40-52 weeks after enucleation, the nerve persists, at 20 40% of the normal diameter, consisting mostly of normal looking astrocytes. The amplitude of the voltage dependent Na currents recorded from nerves during the first 1-4 weeks after enucleation, with the pial sheath intact, decreases by about 50%. After 8 weeks, the Na current recorded from the surface is about 30% of control. At 16-52 weeks after removal of the retina, when there are no intact axons, the Na current is reduced by 90%. If, however, the pial sheath is stripped away, the Na currents recorded from the glial surface are 40-50% of control during this same 16- to 52-week period, suggesting that in the all-glia nerve, the currents are shunted by the relatively thicker pial sheath.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406679 TI - Myelination by mature ovine oligodendrocytes in vivo and in vitro: evidence that different steps in the myelination process are independently controlled. AB - The ability of isolated mature post-myelination ovine oligodendrocytes to myelinate was investigated in tissue culture and in vivo. In culture, although the cells adhered preferentially to rat dorsal root ganglia (DRG) axons, sent out processes that encircled and wrapped them, proliferated, and synthesised myelin proteins (MBP), no myelination was found. This failure to find myelination occurred despite the fact that the oligodendrocytes both in the present experiments and in previous studies elaborated membranous structures that have been shown chemically and structurally to be similar to normal central nervous system myelin. These findings contrasted with those seen when neonatal rodent glial cells were added to similar DRG neuron cultures, in which myelination readily occurred. When the same adult ovine oligodendrocytes were transplanted into the brains of Shiverer mice, normal compact myelin was formed, proving that the cells were capable of myelination and suggesting that cross-species incompatibility was probably not a major factor in the lack of myelination in vitro. It is possible that the failure of ovine oligodendrocytes to myelinate DRG axons is due either to the relatively low number of supporting glial cells, such as astrocytes or microglia which may be necessary for satisfactory myelination, or that some other factor in the microenvironment is lacking; in any event, these results point to the complexity of oligodendrocyte-axon interactions. It is clear that each of the events, from adherence to proliferation to wrapping and the myelin compaction may be under the control of a different signal and may operate through a distinct mechanism, even though each process is dependent on the other. The results also point to the potential usefulness of this model system for deciphering such signals and mechanisms. PMID- 8406680 TI - Insulin-like growth factor I: a mitogen for rat Schwann cells in the presence of elevated levels of cyclic AMP. AB - To develop effective procedures for improving the regeneration of peripheral nerves and for preventing the formation of neurofibromas, it is necessary to identify the different mitogens that stimulate the proliferation of Schwann cells. Insulin-like growth factor I (IGF-I), which is a potent autocrine growth factor in many tissues, is synthesized by proliferating Schwann cells. However, the role of IGF-I in stimulating their division is still uncertain. Here we show that nanomolar concentrations of IGF-I stimulate the growth of Schwann cells in primary culture. IGF-I alone was uneffective, but in the presence of forskolin (5 microM) or dibutyryl cyclic AMP (dbcAMP, 10 microM), it became a potent mitogen. Neither IGF-II nor epidermal growth factor (EGF) were effective even in the presence of forskolin. Insulin also stimulated Schwann cell proliferation in the presence of forskolin, but only at micromolar concentration. Receptors for IGF-I were visualized on the Schwann cell surface by indirect immunofluorescence staining using anti-human IGF-I receptor antibodies. Their presence was also assessed by binding assays using [125I]-IGF-I as a ligand. Scatchard analysis showed a single class of high-affinity receptors (Kd = 1.5 nM). Competitions studies with unlabeled IGF-I or insulin indicated a half-maximal displacement of [125I]-IGF-I by IGF-I at about 5 nM, while insulin was about 500-fold less effective. The number of binding sites for IGF-I was increased by exposing cells for 3 days to forskolin (-forskolin: about 5,100; + forskolin: about 12,200 binding sites/cell).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406681 TI - Intracellular pH regulation in single cultured astrocytes from rat forebrain. AB - We used the fluorescent pH-sensitive dye 2',7'-bis(carboxyethyl)-5,6 carboxyfluorescein (BCECF) to monitor intracellular pH (pHi) in single astrocytes cultured from the forebrain of neonatal rats. When exposed to a nominally CO2/HCO3(-)-free medium buffered to pH 7.40 with HEPES at 37 degrees C, the cells had a mean pHi of 6.89. Switching to a medium buffered to pH 7.40 with 5% CO2 and 25 mM HCO3(-) caused the steady-state pHi to increase by an average of 0.35, suggesting the presence of a HCO3(-) -dependent acid-extrusion mechanism. The sustained alkalinization was sometimes preceded by a small transient acidification. In experiments in which astrocytes were exposed to nominally HCO3( )-free (HEPES-buffered) solutions, the application and withdrawal of 20 mM extracellular NH4+ caused pHi to fall to a value substantially below the initial one. pHi spontaneously recovered from this acid load, stabilizing at a value approximately 0.1 higher than the one prevailing before the application of NH4+. In other experiments conducted on cell bathed in HEPES-buffered solutions, removing extracellular Na+ caused pHi to decrease rapidly by 0.5. Returning the Na+ caused pHi to increase rapidly, indicating the presence of an Na(+) dependent/HCO3(-)-dependent acid-extrusion mechanism; the final pHi after returning Na+ was approximately 0.08 higher than the initial value. This pHi recovery elicited by returning Na+ was not substantially affected by 50 microM ethylisopropylamiloride (EIPA), but was speeded up by 50 microM 4,4' diisothiocyanostilbene-2,2'-disulfonate (DIDS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406682 TI - Receptor-mediated phospholipase D activity in primary astroglial cultures. AB - Phospholipase D, an enzyme involved in signal transduction cascades, catalyses the formation of phosphatidic acid and, when ethanol is present, the formation of phosphatidylethanol. In the present study we demonstrate that stimulation of muscarinic acetylcholine receptors as well as P2-purinergic receptors induces activation of phospholipase D in primary cultures of astroglial cells. Both the hydrolysis and the transphosphatidylation reactions were stimulated by receptor agonists. Carbachol and ATP induced a rapid increase in the amount of [3H]phosphatidic acid in astroglial cells prelabelled with [3H]oleic acid. When ethanol (150 mM) was present, phosphatidylethanol was formed. Furthermore, the receptor-mediated increase in the concentration of phosphatidic acid was inhibited by ethanol, indicating that the phosphatidic acid production was indeed mediated by phospholipase D. The formation of phosphatidylethanol was concentration dependent, with a half-maximal effective concentration of 5 x 10( 5) M for carbachol and 10(-5) M for ATP. The carbachol-induced response was almost completely inhibited by atropine, indicating activation of phospholipase D via muscarinic receptors. The purinergic response is most probably mediated via P2-receptors since ADP was almost as efficient as ATP in inducing phosphatidylethanol formation, whereas AMP was significantly less potent. We conclude that astroglial cells in primary culture display muscarinic and purinergic receptors coupled to phospholipase D. The relationship to cell function needs to be further investigated. PMID- 8406683 TI - Axons modulate the expression of transforming growth factor-betas in Schwann cells. AB - We have investigated the expression of transforming growth factor (TGF)-beta 1, beta 2, and -beta 3 in developing, degenerating, and regenerating rat peripheral nerve by immunohistochemistry and Northern blot analysis. In normal adult sciatic nerve, TGF-beta 1, -beta 2, and -beta 3 are detected in the cytoplasm of Schwann cells, and the levels of TGF-beta 1 and -beta 3 mRNAs are constant during post natal development. When sciatic nerves are transected to cause axonal degeneration and prevent axonal regeneration, the level of TGF-beta 1 mRNA in the distal nerve-stump increases markedly and remains elevated, whereas the level of TGF-beta 3 mRNA falls modestly and remains depressed. When sciatic nerves are crushed to cause axonal degeneration and allow axonal regeneration, the level of TGF-beta 1 mRNA initially increases as axons degenerate, and then falls as axons regenerate. TGF-beta 2 mRNA was not detected in developing or lesioned sciatic nerves at any time. Cultured Schwann cells have high levels of TGF-beta 1 mRNA, the amount of which is reduced by forskolin, which mimics the effect of axonal contact. These data demonstrate that Schwann cells express TGF-beta 1, -beta 2, and -beta 3, and that TGF-beta 1 and -beta 3 mRNA predominate over TGF-beta 2 mRNA in peripheral nerve. Axonal contact and forskolin decrease the expression of TGF-beta 1 in Schwann cells. PMID- 8406684 TI - Differential activation of microglia and astrocytes in aniso- and isomorphic gliotic tissue. AB - Reactive astrocytes and microglial cells are both involved in the formation of gliotic tissue. Using immunohistochemical markers, we have compared the response of both these cell types after two different kinds of damage in the brain: traumatic injury (anisomorphic gliosis) and neurotoxic induced lesion (isomorphic gliosis), in two distinct regions of the brain, the cortex and the hippocampus. We show that the time course and the relative contribution of astrocytes and microglial cells differ greatly in the two kinds of lesions. While in anisomorphic gliosis there is little activation of endogenous microglial cells independently of the brain region damaged, these cells contribute in large measure and for prolonged periods of time to the formation of isomorphic gliotic tissue. Astrocytes are quickly activated at the border of anisomorphic lesions, and after 3 days they already occupy an extensive portion of the brain parenchyma. However, after 1 month, they are found restricted to a thin strip at the lesion boundary. In contrast, after an isomorphic lesion, astrocytes become reactive around the site of neuronal cell loss but not at the site of the lesion itself. Only after 2 weeks do they totally invade the damaged region, persisting for at least 1 month. Such differences are observed independently of the brain region damaged. These results suggest that the cellular, and therefore the molecular, composition of gliotic tissue depends on the type of insult the CNS has suffered. PMID- 8406685 TI - Differential effects of reactive oxygen species on native synovial fluid and purified human umbilical cord hyaluronate. AB - The ability of reactive oxygen species produced by triggered neutrophilic leukocytes, hypoxanthine/xanthine oxidase (HX/XAO), hydrogen peroxide, and hypochlorous acid/myeloperoxidase (HOCl/MPO) systems to degrade hyaluronate (HA) in human synovial fluid (SF) and purified umbilical cord HA was compared by measuring the molecular weight distribution of HA using high-performance liquid chromatography with a size-exclusion column. The exposure of noninflammatory SF to phorbol myristic acetate (PMA)-activated neutrophils or to hydrogen peroxide (H2O2) caused depolymerization of SF HA to the degree corresponding to that found in rheumatoid SFs. When HX/XAO was used as radical generator, the molecular weight of SF HA decreased from 3.42 x 10(6) to 1.40 x 10(4) daltons with concomitant decrease of SF viscosity to 36% from the original value. The HOCl/MPO system caused no depolymerization of SF HA, even at very high unphysiological HOCl concentrations that induced the precipitation of SF HA together with SF proteins. This effect was found to be comparable to conventional mucin clot formation in SF. However, purified human umbilical cord HA was easily depolymerized with HOCl/MPO or with H2O2, but these effects were sensitive to the hydroxyl radical scavenger mannitol and iron chelator desferrioxamine, indicating that the formation of reactive hydroxyl radical (OH.) is likely to participate in these reactions. Thus we conclude that in inflammatory SF HA is mainly depolymerized by OH. produced by decomposition of H2O2 catalyzed by iron, free or locally bound to HA itself. In contrast to what has been reported earlier, HOCl/MPO only depolymerizes purified umbilical cord HA (in a hydroxyl radical dependent manner) but does not depolymerize HA in SF. As a matter of fact, HOCl/MPO has a scavenging action on SF HA by consuming H2O2 and thus preventing the formation of reactive hydroxyl radicals. PMID- 8406686 TI - Comparison of in vitro effects of flunixin and tolfenamic acid on human leukocyte and platelet functions. AB - A study was made to compare the effects of two nonsteroidal antiinflammatory drugs (NSAIDs), flunixin and tolfenamic acid, on the leukotriene B4 (LTB4) production and migration of human polymorphonuclear leukocytes (PMNs) as well as on platelet aggregation and thromboxane B2 (TxB2) production during blood clotting. Tolfenamic acid inhibited LTB4 production in PMNs as well as FMLP- and LTB4-induced PMN migration (IC50 values 23 +/- 3, 39 +/- 11, and 68 +/- 13 microM, respectively), whereas flunixin inhibited these cell functions only with the highest concentration tested (100 microM). On the other hand, flunixin was clearly a more potent inhibitor of TxB2 production and adrenaline-induced platelet aggregation than tolfenamic acid, the IC50 values in TxB2 production being 0.28 +/- 0.02 microM and 2.6 +/- 0.3 microM for flunixin and tolfenamic acid, respectively. We suggest that inhibition of PMN functions may be an additional mechanism in the antiinflammatory action of tolfenamic acid. At least in human PMNs and platelets, flunixin seems to be only an inhibitor of cyclooxygenase. PMID- 8406687 TI - Functions of polymorphonuclear leukocytes in early rheumatoid arthritis. AB - We carried out a prospective study on clinical variables and functions of polymorphonuclear leukocytes (PMNs) of 20 patients with early rheumatoid arthritis (RA) and compared the results with the presence of erosions before treatment and at a one-year follow-up. Migration of PMNs determined by agarose and filter assays and respiratory burst of PMNs determined by luminol-enhanced chemiluminescence (CL) test were studied both before starting RA-modifying treatment and 6-12 (mean 7.3) months later. PMNs of the patients without erosions at one year, as compared to the patients with erosions, showed significantly depressed migration into filter and significantly depressed CL responses to N formyl-methionyl-leucyl-phenylalanine, both before starting the treatment and at 7.3 months. Although causality remains uncertain, the results suggest that depressed functional capacity of PMNs is associated with low risk of joint destruction in early RA. PMID- 8406688 TI - The magnitude of macrophage inflammatory response does not directly depend on ability of bone marrow cells to respond to interleukin-3 in mice of different strains. AB - The multicolony stimulating factor Interleukin-3 (IL-3) has a role in regulating the proliferation, differentiation, and survival of myeloid stem cells and committed progenitor cells within each of the myeloid lineages. It has been referred to as an emergency factor appearing following triggering of an inflammatory response. The ability of bone marrow (BM) stem cells to respond to a stimulus such as IL-3 in vitro may reflect the in vivo capacity of BM stem cells to generate newly BM-derived macrophages being recruited to an inflammatory site. Both parameters, namely the BM cell response to in vitro IL-3 treatment and the magnitude of the macrophage inflammatory response vary among inbred mouse strains. Mice of the A/J strain are known to have weak macrophage inflammatory response to a phlogistic agent and their BM cells are hyporesponsive to IL-3 exposure. In contrast, mice of the C57BL/6 strain mount a high macrophage inflammatory reaction to a stimulus, and their BM cells strongly proliferate in response to the presence of IL-3. Thus, we examined whether or not the type of BM cell response to IL-3 (i.e., A/J- or C57BL/6-like) determines the magnitude of the macrophage inflammatory response using the A x B/B x A recombinant inbred (RI) mouse strain system. The two traits were found not to cosegregate, suggesting that they are not linked. The continuous strain distribution pattern of the magnitude of the macrophage inflammatory response obtained in mice of the A x B/B x A RI strains implies that this trait is under the control of several genes. PMID- 8406689 TI - Distribution of pancreatic (group I) and synovial-type (group II) phospholipases A2 in human tissues. AB - The catalytic activity of phospholipase A2 (cat-PLA2) and the concentration of pancreatic (group I) phospholipase A2 (pan-PLA2) and synovial-type (group II) phospholipase A2 (syn-PLA2) were studied in 19 human tissues in order to find potential sources of circulating phospholipase A2. Five specimens of each tissue were collected at autopsies or from normal deliveries (placentas and amnionic membranes). The concentrations of pan-PLA2 and syn-PLA2 were measured by specific time-resolved fluoroimmunoassays. The cat-PLA2 was measured by radioactive dipalmitoyl phosphatidylcholine as substrate. The concentration of pan-PLA2 was negligible in all tissue homogenates except the pancreas. Immunoreactive syn-PLA2 was found in the homogenates of the digestive tract, cartilage, and prostatic and parotid glands. By immunohistochemistry, syn-PLA2 was localized in Paneth cell secretory granules, chondrocytes, cartilage matrix, and glandular cells of prostate. PMID- 8406690 TI - Cytokines increase proliferation of human intestinal smooth muscle cells: possible role in inflammation-induced stricture formation. AB - Crohn's disease is an idiopathic, chronic inflammation of the gastrointestinal tract that causes narrowing and stricturing of primarily the small and large intestine. Although the mechanism(s) by which chronic inflammation promotes stricture formation remain to be defined, it does appear to be associated histologically with a hyperplasia of smooth muscle cells and an increased deposition of collagen within the bowel wall. The objective of this study was to assess the effect of two proinflammatory cytokines, tumor necrosis factor and interleukin-1, on the proliferation of human intestinal smooth muscle cells in vitro. Human intestinal smooth muscle cells were seeded at subconfluent densities into 24-well plates in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. Human recombinant tumor necrosis factor (0.1-100 ng/ml), interleukin-1 (0.1-500 ng/ml), or control medium (without cytokines) was then added to the cells and incubation continued for 48 or 72 h. Proliferation was determined by the incorporation of tritiated thymidine, added during the final 18 h, into the cellular DNA of the smooth muscle cells. Both cytokines caused a significant dose-dependent increase in intestinal smooth muscle cell proliferation relative to control. These results suggest that the interleukin-1 and tumor necrosis factor produced during chronic inflammation in vivo may enhance the proliferation of smooth muscle cells within the intestinal bowel wall and hence potentially contribute to the narrowing and stricturing of the intestine that is observed in Crohn's disease. PMID- 8406691 TI - Acetylsalicylic acid and misoprostol combination in adjuvant arthritis of rats. AB - The combined effect of acetylsalicylic acid (ASA) and misoprostol (MISO) on adjuvant arthritis was investigated on rats. Alteration by various doses of MISO and fixed dose of ASA was studied. Drugs were given by the nasogastric route each day beginning from the day of adjuvant injection (day 0) and continued until the 16th day. Paw swelling was measured on days 4, 17, and 29, and secondary lesions were assessed on days 17 and 29. Pathological examination of tibiodorsal junction was also evaluated on the 29th day. The results clearly showed that the combination of MISO with ASA did not inhibit the antiinflammatory effect of ASA. Unexpectedly, MISO increased the antiinflammatory effect of ASA at some dosage regimens. PMID- 8406692 TI - Presence of serum modulates expression of complement receptor type 1 (CR1) on human granulocytes after quartz exposure. AB - We have investigated the interaction between granulocytes and quartz with respect to the expression of complement receptor type 1 (CR1) and the presence of normal human serum (NHS). Quartz down-regulates selectively CR1 on activated granulocytes. This down-regulation is abolished in the presence of both NHS and heat-inactivated NHS (NHS56) but not human albumin. When quartz was preincubated with NHS (quartz-NHS) before exposure to activated granulocytes, a down regulating effect was observed in contrast to preincubation with NHS56, which did not induce a down-regulation. Preincubation with cytochalasin B reduced the down regulation of quartz-NHS, indicating a cytoskeleton-dependent internalization of the receptor. The serine protease inhibitor PMSF partly reduced this down regulation. Our results indicate that the presence of NHS in the alveolar space influences the interaction between quartz and recruited granulocytes with respect to CR1 expression. Since CR1 is an important opsonin receptor and soluble CR1 can modulate the inflammatory response, this may be of importance in the inflammation and fibrosing process induced by quartz in the alveolar space and lung interstitium. PMID- 8406693 TI - Phorbol myristate acetate induction of chemotactic migration of human polymorphonuclear neutrophils. AB - The in vitro migration of human polymorphonuclear neutrophils (PMNs) was studied employing an enzymatic assay of cell migration with phorbol myristate acetate (PMA) as the test stimulant. Our data clearly show that PMA in concentrations between 1 and 100 ng/ml in the lower wells of blind-well chambers induced chemotactic migration. Chemokinesis (increased migration) was not induced when PMA was present in both the upper and lower chambers (i.e., in a nongradient mode). Clearly our data indicate that PMA is chemotactic for human PMNs and, coupled with published studies of the effect of PMA on PMNs, suggest activation of an intracellular gradient of membrane-associated protein kinase C as a possible new mechanism for the induction of oriented migration of PMNs. Such a mechanism may be generalized to include membrane-soluble materials (e.g., inflammatory mediators, microbial products), which establish internal gradients of activated PKC rather than via the "classic" agonist-surface receptor mechanism, providing an alternative pathway for the induction of leukocyte chemotaxis. PMID- 8406694 TI - Liquid acute epidural hematoma. PMID- 8406695 TI - Duplicate perineal anus. A rare anorectal malformation. PMID- 8406696 TI - Hydrocephalus presenting as precocious puberty. PMID- 8406697 TI - HIV serosurveillance in multi-transfused thalassemic children. PMID- 8406698 TI - Fixed drug eruption in infancy. PMID- 8406699 TI - Training of final year MBBS students in neonatal resuscitation. AB - Sixty final year MBBS students were trained in the art of neonatal resuscitation based on the 'NALS' course material. The students scored 11.6 +/- 3.02 in the pretest and 17.85 +/- 4.42 (out of 20) in the post-test. The trainees felt that the programme was extremely useful. Majority of them said that the course content was optimum with appropriate use of teaching-learning media. It is recommended that this training may be imparted to all the undergraduate students at the entry of final year MBBS course before their labor room posting so that they can have effective reinforcement of the training. PMID- 8406700 TI - Oxygen therapy in pediatric practice. PMID- 8406701 TI - Comparative outcome of low birth weight babies. AB - One hundred and fifty six babies with birth weight between 1500-2000 g and 103 full term-appropriate for gestational age (FT-AGA) babies delivered at University Hospital, District Hospital and village homes were included for a comparative study of mortality, morbidity and growth pattern. The low birth weight (LBW) babies from the three centres had similar birth weight and gestational age. Neonatal mortality rates for the LBW babies were similar at the three centres. The main cause of death were infections and aspiration with rates again being similar. Diarrhea and respiratory tract infections were common causes of morbidity. The mortality rates for the LBW babies were significantly higher as compared to FT-AGA babies irrespective of the place of delivery. The incidence of morbidities like diarrhea and respiratory infections were also higher in LBW babies. However, the differences were statistically significant mostly in the preterm group. The weight gain of all LBW babies was similar up to 3 months of age. The findings of an identical outcome for the LBW babies at village level to those managed at hospitals is an encouraging trend to increasing domiciliary care for LBW babies. PMID- 8406702 TI - Foot tape measure for identification of low birth weight newborns. AB - The majority of births in rural India take place at home, Logistic constraints make early and reliable identification of low birth weight babies difficult. Using neonatal foot length as a proxy measure for birth weight, we devised a tri colored foot tape intended for use at home by the neonatal caretaker or birth attendant. The tape was field tested in a rural community in the Pune district. Results showed a sensitivity of 68.2% and a predictive value of 45.5% for identifying low birth weight. For very low birth weights (< 1500 g) the sensitivity was 100%, specificity 95.2% and the positive predictive value 60%. Interobserver reliability comparing a trained medical social worker and the household member was high (kappa score of 0.82). If implemented on a larger scale this simple, low cost technology has the potential to significantly enhance the yield of identification of low birth weight babies born at home. PMID- 8406703 TI - Ontogeny of stool passage in low birth weight infants < or = 1500 grams. AB - The onset of stool passage, timing of transition to yellow stools and the pattern of stooling frequency over the first 4 weeks were studied in infants < 1500 g at birth. The time of passage of the first stool (median, 19) correlated with birth weight and gestational age but not with presence or severity of respiratory distress; fourteen percent passed stool after 1st 48 hours. Transition to yellow occurred at 17.6 +/- 6.4 days and was related to the onset of feeding and birth weight. Stooling frequency was similar in Wk 2 as Wk 1, increased in Wk 3 and plateaued on Wk 4. Volume of feeding/day increased each week over that of preceding week but stooling frequency was not related to the increased volume or any of the other variables. PMID- 8406704 TI - Comparison of rectal and axillary temperatures in neonates admitted to a neonatal unit. AB - To determine whether axillary temperature could be used as an alternative to rectal temperature, 45 neonates were subjected to simultaneous rectal temperature and axillary temperature recordings. Each baby had an average of 10 recordings over the first 72 hours. The mean of these readings was taken and subjected to regression analysis and the 't' paired test. The overall mean difference between the rectal and axillary temperature was 0.3 degrees F. The mean difference was least in preterm small for date (SFD) babies (0.2 degrees F) and most in preterm appropriate for date (AFD) babies (0.5 degrees F). The mean difference was less in incubator babies (0.2 degrees F) compared to those in warmer cradles (0.4 degrees F). The regression analysis showed that the correlation was good in all groups except the term small for date babies. We conclude that axillary temperature could be used as an alternative to rectal temperature in neonates. PMID- 8406705 TI - Percentile growth charts for head circumference in Punjabi infants. AB - Percentile growth charts for head circumference of Punjabi infants (male 86 and female 68) measured serially at monthly intervals during the first year of life are presented. Besides, constancy of sample size and strict adherence to time tolerance limit of +/- 3 days, on the day of monthly measurements other factors considered responsible for the natural smooth course of the percentile grids are highlighted. The presented data would enable easy detection of infants with abnormal course of head growth. PMID- 8406706 TI - Clinical profile and outcome in enteric fever. AB - Sixty five blood culture positive cases of S. typhi were studied for clinical profile. A total of 64.6% were multidrug resistant and 35.4% were chloramphenicol sensitive. In patients with multidrug resistant S. typhi the age was higher (p < 0.01), and incidence of complications such as shock (35.7%), encephalopathy (42.9%), myocarditis (14.3%) and gastric hemorrhage (4.7%) were more frequent, compared to chloramphenicol sensitive group. Cases with multidrug resistant S. typhi (MDRST) were treated with oral ciprofloxacin; the period of defervescence of fever was significantly less (p < 0.05) compared to the chloramphenicol group. Our study suggests the use of ciprofloxacin in the treatment of MDRST without any side effects. PMID- 8406707 TI - Meconium aspiration syndrome: recent concepts. PMID- 8406708 TI - Prevalence of rheumatic heart disease in school children. AB - A total number of 8449 school children, in the 5-15 years old, group were examined clinically for evidence of valvular lesions and confirmed by roentgenographic electrocardiographic and echocardiographic findings. Twelve children (0.14%) were suffering from rheumatic heart disease (RHD). The disease was significantly more (p < 0.05) prevalent in the higher age group of 11-15 years as compared to younger children. Boys (0.18%) were affected more than girls (0.09%). The disease was more frequent among children belonging to economically backward group (0.34%) and those living in large families (0.28%). Children living in the slums had significantly (p < 0.05) higher prevalence (0.41%) than those residing in urban areas (0.06%). Only 3 out of 12 (25%) were previously diagnosed cases and remaining were diagnosed for the first time. Past history of rheumatic activity was seen in 41.67% cases. Mitral valve involvement was most frequent, being detected in 10 (83.33%) cases. Majority of cases (58.33%) had the development of symptomatic RHD within 2 years of having suffered from RF. Our study helped to detect the hitherto undiagnosed cases of RHD and showed that its prevalence was higher in older children and those from urban slums. PMID- 8406709 TI - Pattern of childhood trauma. Indian perspective. AB - Although infectious diseases are still the chief cause of death in children in a developing country like ours, but a definite increase in incident and related mortality due to trauma has been noted in the last decade. The problems relating to pediatric trauma are peculiar to our setup and differ considerably to the severe multiple organ trauma met with in the high velocity vehicular accidents, seen in developed Western countries. The present study identifies patterns of childhood trauma from our region. It comprises 2100 patients admitted over a 3 years period to Pediatric Surgical Unit. Cranial injuries were the most commonly encountered injuries followed by the abdominal and skeletal injuries. Fall from the house roofs is the commonest mode of injury, although road traffic accidents are also recognized to be on the increase. The overall mortality was 7.7%; its chief determinant being the presence of severe head injury. A few important epidemiological factors involved were identified. PMID- 8406710 TI - Proptosis: management of 22 patients. AB - The clinical features and management of 22 cases with proptosis is highlighted. Proptosis was unilateral in fifteen and bilateral in seven cases. The common etiologic factors were neoplasms, infections and bleeding diatheses. Of the ten cases with neoplasms as many as 7 expired, whereas of nine cases due to infections only one expired and one developed phthisis bulbi. All three patients with proptosis due to bleeds recovered completely without sequelae. Orbital CT scan was done in fourteen and ultrasound in eight cases, while in five cases both investigations were done. As neoplasms are a frequent cause of proptosis in children and there is a high mortality in this group, a thorough systemic examination and hematological profile is essential to arrive at a speedy diagnosis. CT and USG are useful imaging modalities of the orbit, the latter being particularly useful for follow up of the lesion. PMID- 8406711 TI - Ceftriaxone: use in multidrug resistant typhoid fever. PMID- 8406712 TI - Antiphospholipid antibodies (APA) and cerebral stroke. PMID- 8406713 TI - Carbamazepine therapy for infantile tremor syndrome. PMID- 8406714 TI - Amphotericin B in visceral leishmaniasis. PMID- 8406715 TI - Phenytoin toxic encephalopathy. PMID- 8406716 TI - Acute digoxin toxicity in a neonate. PMID- 8406717 TI - Nifedipine in urticaria. PMID- 8406718 TI - Sublingual terbutalin in bronchial asthma. PMID- 8406719 TI - Maternal knowledge, attitudes and practice in diarrhea. PMID- 8406720 TI - De Sanctis Cacchione syndrome. PMID- 8406721 TI - Superficial fungal infections in newborns. PMID- 8406722 TI - Management of meconium stained amniotic fluid: a team approach. AB - Passage of meconium in utero is a serious neonatal disorder carrying high morbidity and mortality. Role of planned team approach with aggressive intrapartum suctioning and intensive neonatal management was studied to evaluate its impact on neonatal morbidity and mortality. Meconium Stained Amniotic Fluid (MSAF) was found amongst 7.4% of all deliveries and among these 238 (10.5%) babies developed meconium aspiration syndrome (MAS). Ninety five per cent babies with MSAF were born at > 36 weeks of gestation and 76% were more than 2.5 kg. Passage of thick and thin meconium was seen in 44 and 56% respectively. Passage of thick meconium was significantly associated with severe asphyxia and carried a bad prognosis with increased risk of development of meconium aspiration syndrome, hypoxic schemic encephalopathy, seizures and pulmonary air leak syndrome. Aggressive team approach was responsible for lowering the mortality to 7.7%. PMID- 8406723 TI - Freeman Sheldon syndrome with bilateral simian crease and malpositioned second toes. PMID- 8406724 TI - Askin Rosai tumor. PMID- 8406725 TI - Childhood appendicitis. A clinical profile. AB - A study to assess the reliability of clinical symptoms and signs in 50 patients with a presumptive diagnosis of acute appendicitis is presented. The male to female ratio was 3: 2, with age ranging from 2 to 15 years. Abdominal pain was present in 42; tenderness was localized in 35, generalized in 11 and diffuse in 4 patients. Total leucocyte count was above 11,000/cu mm in 31, below 11,000/cu mm in 17 and above 18,000/cu mm in 2. Of the 48 operated patients, 8 had normal appendices and the diagnosis in them was Meckel's diverticulitis 3, ruptured ovarian follicle 2, mesenteric adenitis 2, and salpingo-oophoritis 1. Abdominal pain and right iliac fossa tenderness with contributory investigations are the most reliable indicators of acute appendicitis with a false positive rate of 16.66% only. PMID- 8406726 TI - Approach to the adult patient with acute diarrhea. AB - Acute diarrhea in the adult is caused primarily by acute infectious organisms, mainly viruses but also bacteria and parasites. The majority of cases are self limited and resolve without sequelae or specific intervention. Diagnostic evaluation should be restricted to those with severe symptoms such as significant volume depletion or dysentery or who are altered hosts due to immunosuppression or chronic illness. Diagnostic testing in less ill patients with probable viral illnesses is unrevealing and wasteful of patient resources. Therapy consists simply of correcting the major harm caused by organisms: volume depletion. Specific therapy with antibiotics should be restricted to situations in which proven efficacy has been demonstrated. The approach to the patient with acute infectious diarrhea is strictly a clinical one, with a careful and thoughful ordering of diagnostic tests in cases in which the information is likely to change management and outcome for the patient. PMID- 8406727 TI - Approach to the pediatric patient with diarrhea. AB - Numerous viral, bacterial, and parasitic pathogens are known to cause diarrheal illnesses with increased frequency in children. Oral rehydration can be used to treat and prevent dehydration, the major sequela of diarrhea in children. The impact of diarrhea on nutrition may also be reduced through the rapid restoration of a normal, age-appropriate diet. Most diarrheal illnesses are acute and self limited; however, increased knowledge of persistent diarrheal syndromes in children may lead to prompt recognition and diagnosis in children with diarrhea lasting more than 2 weeks. PMID- 8406728 TI - Approach to acute diarrhea in the elderly. AB - Diarrhea is a common problem among the elderly that can have catastrophic results. Atherosclerosis predisposes older adults to morbid sequelae from dehydration resulting from diarrhea. Deaths related to diarrheal illnesses are recognized among older adults living in the community as well as among those confined to nursing homes. Outbreaks have most often been associated with excess deaths from diarrhea among nursing-home patients. Although most cases of dehydration from diarrhea result from gastrointestinal infections, noninfectious causes of diarrhea related to prescription of laxatives, side effects of medications, and use of enteral feedings are common. Clostridium difficile infection is particularly common among older adults in hospitals and nursing homes, and relapsing disease in these groups may be more frequent than among younger adults. The approach to an elderly patient with diarrhea is to ensure proper hydration using available oral rehydration solutions, proceed with diagnostic tests likely to yield a positive result, avoid the use of harmful antiperistaltic drugs, and provide adequate follow-up of the nutritional state. PMID- 8406729 TI - Infectious diarrheas in patients with AIDS. AB - In conclusion, diarrheal illnesses remain a major clinical problem for patients infected with HIV-1, particularly those with AIDS. Opportunistic enteric pathogens for which there is no effective treatment, the emergence of new opportunistic agents, and the enlarging patterns of drug resistance continue to challenge us. However, better understanding of HIV-1-induced mucosal immunosuppression, sound clinical judgement, careful diagnostic evaluation, development of newer antimicrobial agents, and judicious patient management should help us meet this challenge. PMID- 8406730 TI - Traveler's diarrhea. AB - A variety of infectious enteric pathogens (bacterial, viral, and protozoal) can lead to a systemic diarrheal illness in international travelers traveling from industrialized countries to developing areas of the world. Many of the agents that lead to this syndrome have been identified, and their mode of transmission has been defined. Prophylactic measures are advisable, and effective treatment options are available. This article also discusses issues important in the management of patients who develop a chronic diarrheal illness after travel. PMID- 8406731 TI - Diarrhea acquired in the hospital. AB - Nosocomial diarrhea may be the second or third most common nosocomial infection, depending on the type of institution and the presence of outbreaks. Age is a major determinant of the type of cause of the diarrhea and, as a consequence, the choice of treatment. Control of nosocomial outbreaks is usually linked to prompt diagnosis and treatment of individual cases, interruption of the route(s) of transmission, and an increase in the awareness of infection-control procedures. Unfortunately, the complete elimination of nosocomial diarrhea is not feasible, and these infections will remain a challenge to us all in the future. PMID- 8406732 TI - Food poisoning syndromes. AB - Despite our society's advances in sanitation, food preservation, and hygiene, the prevalence of foodborne disease remains high (12.6 million cases per year in the United States). Although there is a constant need for education of food handlers and consumers, there is also a need for continued vigilant monitoring of coastal waters, meat packing facilities, and imported foods. As long as antibiotics are used in poultry and cattle feeds, one can expect the incidence of antibiotic resistant foodborne pathogens to rise. There are several promising areas of research in the field of foodborne illnesses. Molecular biologists are actively characterizing the genes that enable invasive enteric pathogens such as Salmonella and Yersinia to enter tissues, and the bacterial toxins associated with secretory diarrheas continue to be the subject of intense scrutiny. Epidemiologists are implementing new techniques such as DNA fingerprinting and multilocus enzyme electrophoresis for tracing pathogens in disease outbreaks. Similarly, the use of computers in the food industry facilitates the tracing of contaminated foods. Although the rates of foodborne illness may not decrease significantly during the next decade, we can expect more rapid identification and tracing of outbreaks as well as an improved understanding of the pathogenesis of the foodborne diseases. PMID- 8406733 TI - Escherichia coli diarrhea. AB - E. coli diarrheal disease is becoming ever more complicated as more and more pathogenic mechanisms are identified. E. coli strains remain the major causes of infectious diarrhea worldwide. Presumptive diagnosis based on clinical and laboratory criteria is practical for strains known to be important in the United States. Specific diagnosis is not currently feasible outside of research centers. Therapy, when indicated, shortens the duration of illness. Research is proceeding rapidly at the molecular level and may lead to new diagnostic and therapeutic approaches in the near future. PMID- 8406734 TI - Clostridium difficile colitis and diarrhea. AB - Clostridium difficile is now regarded as the most prevalent nosocomial pathogen, infecting as many as a quarter of hospitalized patients. The pathophysiology of infection with this unusual enteric pathogen involves alteration of the normal enteric flora by antibiotics, ingestion of spores, and colonization by C. difficile. The organism then releases potent exotoxins that produce an inflammatory colitis and diarrhea. A spectrum of host responses occurs, ranging from the asymptomatic carrier state to life-threatening pseudomembranous colitis. Effective therapy with vancomycin or metronidazole is available, but relapses occur in 15% to 20% of patients and may necessitate multiple courses of therapy. PMID- 8406735 TI - Modern diagnosis (with molecular tests) of acute infectious diarrhea. AB - There have been three significant technical developments in the molecular genetic diagnosis of infectious diarrhea. The first was the replacement of polynucleotide probes with more specific synthetic oligonucleotide probes. The second was the replacement of radiolabeled markers with nonradiolabeled markers, and the third was PCR amplification. In the PCR procedure, it is possible to increase the quantity of target nucleotide sequences to quantities easily detectable with nonradioactive oligonucleotide probes. It is now possible to amplify nucleotide sequences of more than one enteric pathogen with different primers simultaneously and to detect these amplified nucleotide sequences with nonradiolabeled oligonucleotide probes. With a scanning laser system, the results of enteric PCRs will be used to identify enteric pathogens on a routine basis in clinical and public health laboratories. Computers need to be used to analyze these results and transfer this information rapidly to clinicians and public health officials. PMID- 8406736 TI - Nonantibiotic therapy and pharmacotherapy of acute infectious diarrhea. AB - The most important aspect of nonantibiotic treatment of diarrhea is to prevent or repair dehydration and thus prevent significant morbidity or death. Although intravenous fluid therapy is effective, it is expensive and requires personnel and equipment. ORT has saved millions of lives in emerging nations and should be more commonly used in developed countries. Although the simple concept of glucose stimulated Na+ and fluid absorption remains the basic tenet of such ORT, grain based solutions appear to have added efficacy and nutritional value. Mild, nondysenteric diarrheas can be safely and reasonably effectively treated either with loperamide or with bismuth subsalicylate compounds. Effective adjunctive therapy for severe secretory diarrheas has thus far escaped discovery. Somatostatin analogues such as octreotide have approximately 50% efficacy in reduction of the moderate secretory diarrheas such as those present in AIDS. There are pharmacologic leads that should be explored to develop more gut specific antisecretory forms of alpha 2-adrenergic agents, somatostatin, or phenothiazines. In addition, Cl- channel blockers still hold great theoretic promise. Effective antisecretory therapy has been sought for the past 20 years. It is disappointing that no effective new drugs have yet emerged. Undoubtedly, this speaks of our inexact understanding of the mechanisms that result in secretory diarrheas. Because of the continued loss of life from diarrhea in emerging nations, better understandings of these mechanisms and effective therapies remain appropriate goals for medical science. PMID- 8406737 TI - Rehabilitation after trauma. PMID- 8406738 TI - Multiple trauma in elderly patients. Factors influencing outcome: importance of aggressive care. AB - From 1978 to 1991, 126 multiply-injured patients of 65 years and over were admitted to the Department of Traumatology and Emergency Surgery of the University Hospitals of Leuven. The seriousness of the injury was evaluated using the Injury Severity Score (ISS) and the Glasgow Coma Scale (GCS). Traffic accident (57 per cent) and a simple fall at home (30 per cent) were the main causes of injury. The overall mortality rate within 6 months was 17 per cent. Multiple system organ failure (MSOF) was responsible for the fatal outcome in 48 per cent of the cases and in 71 per cent of the deaths more than 7 days after trauma. Of the survivors still living at home before injury, 78 per cent were able to go back to their normal surroundings. Survivors were compared with non survivors. There was no significant difference in age or in ISS, nor in pre existing diseases. On the other hand, the GCS was of important prognostic value, both for survival and functional recovery (P < 0.001). Also, the need for early intubation and continued ventilation were predictive of survival (P < 0.001). Nevertheless, this need for respiratory assistance was not an indication for withdrawing support as 9 per cent of the survivors also required endotracheal intubation for 5 days or longer. In our opinion, aggressive trauma care for the elderly is justified. PMID- 8406739 TI - Assessment and communication of conscious level: an audit of neurosurgical referrals. AB - Telephone referrals of 100 head-injured patients to a neurosurgical service were assessed to determine if altered consciousness was adequately described by the referring doctor. Of the patients, 60 per cent were resuscitated, assessed and referred by SHOs. Only 30 per cent of doctors were fully conversant with the Glasgow Coma Scale and 18 per cent were unable to describe altered consciousness. Assessment of motor response was often described inaccurately and this was frequently due to the use of an inappropriate stimulus. The use of a pictorial guide to motor responses and a 'Head Injury Proforma' are suggested as means of improving referral and enhancing audit. The majority of doctors had not received formal training in the assessment of conscious level during their undergraduate or postgraduate training. Training video tapes from neurosurgical units showing standard examination methods and typical responses would allow self-assessment and training. PMID- 8406740 TI - Early cytokine response to multiple injury. AB - Serial measurements of the inflammatory cytokines interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF alpha) were made in four patients with multiple injuries to characterize secretion patterns and to relate them to severity of injury and degree of haemorrhage. IL-6 and TNF alpha were assayed using a sandwich ELISA. Injury scores varied from 9 to 50, and transfusion requirements varied from 7 to 14 units. All patients showed an increase in IL-6 but only one a rise in TNF alpha. There was no consistent pattern in the elevation of IL-6 and there was no relation between cytokine levels, severity of injury, transfusion requirement or ultimate outcome. From this preliminary study it would seem that measurement of IL-6 and TNF alpha is not of value in directing therapeutic decisions or as a predictor of outcome. PMID- 8406741 TI - Treatment of combined brachial plexus and subclavian artery trauma. AB - Five cases of combined brachial plexus root avulsion and subclavian artery trauma are presented. A policy of preoperative myelography and minimal vascular reconstruction in the presence of root disruption is discussed. PMID- 8406742 TI - Influence of magnetic resonance imaging on a knee arthroscopy waiting list. AB - Magnetic resonance imaging (MRI) scans were performed on 155 patients who had been placed on a diagnostic knee arthroscopy waiting list 4 to 18 months previously. In all, 32 per cent were removed from the waiting list following this investigation. A total of 24 patients on the waiting list improved spontaneously, including 13 per cent of those with MRI-demonstrable meniscal tears. MRI had an accuracy of 93 per cent, a sensitivity of 100 per cent and a specificity of 67 per cent using arthroscopy as a standard; the false-positive rate was 7 per cent and the false-negative rate 1 per cent. It is concluded that the use of MRI can reduce the requirements for diagnostic arthroscopy, but that the ideal time for MRI is not clear. PMID- 8406743 TI - Knee radiographs: a substitute for proper clinical examination within the accident and emergency department? AB - Over a 2-month period in an accident and emergency department, the clinical benefits and cost-effectiveness of radiology of the knee after trauma was audited. In all, 178 patients had radiographs as part of their initial assessment, the majority of which were subsequently reported by the radiologist as entirely normal (90.4 per cent of films after a twisting injury and 80.6 per cent after a direct blow). Early and appropriate clinical review by a senior doctor is proposed as a viable alternative to perhaps unneeded radiographs where the physical signs of injury are minimal. The clinical and financial benefits of such a policy are discussed. PMID- 8406744 TI - Long-term results of anterior cruciate reconstruction with the patellar tendon. AB - Thirty patients who had an anterior cruciate reconstruction with a patellar tendon autograph were reviewed at an average of 7.5 years (range 5-11 years) after operation. In all, 27 patients maintained their good/excellent results based on the Lysholm and Gillquist scoring system compared with the review at 2 years; two patients remained fair and one reconstruction failed at an early stage. There was no apparent deterioration of knee function at 7.5 years when compared with those at 2 years. None of the patients had symptoms related to the patellofemoral joint. PMID- 8406745 TI - Acute injuries of the upper dorsal spine. AB - In an effort to define better diagnostic and treatment criteria, we reviewed the cases of 50 patients with injuries of the upper dorsal spine from T2-T9, seen between January 1983 and July 1988. The majority of injuries were caused by road traffic accidents, followed by falls from a height, and train accidents. More than 50 per cent had associated injuries, of which four were missed; 50 per cent had neurological deficits, and the majority of these cases had burst fractures and fracture-dislocations. Treatment was non-operative in the majority of cases, with only four cases undergoing anterior spinal decompression and fusion, and one had posterior instrumentation and fusion. At follow-up (1-3 years), only one patient, a child, had significant deformity. All patients with partial neurological deficit improved, whereas none of those with complete neurological lesions recovered. Five patients had significant back pain requiring analgesics but not significant enough to interfere with their activities. We concluded that most of these injuries are stable and should be treated non-operatively. Surgery is only indicated in cases with partial neurological deficit and significant spinal canal compromise or significant instability. Children should be followed up until maturity to look for late deformity. PMID- 8406746 TI - Fracture of the femoral neck: identification of the optimal screw position by migration ratio. AB - A consecutive series of fractures of the femoral neck treated with sliding screw and plate osteosynthesis were evaluated radiographically for the optimal position of the screw by use of migration indices. A naturally occurring optimal position of the screw was discovered. It was characterized by four positional indices in the head-neck region which were significantly correlated with minimal migration (or best stability). This optimal screw position gave a better prognostication of reduced risk of early fracture failure compared with the usual sectional division of the femoral head as indicator of screw position. PMID- 8406747 TI - Titanium staple and screw fixation for recurrent dislocation of the shoulder. AB - A small titanium staple with a locking screw was designed in 1979 to hold the anterior capsule of the shoulder firmly to the front of the scapula just medial to the glenoid. The posterior or inferior capsule can also be stabilized. Two indentations on the staple diminish capsular compression, while a central hole for a 4.5-mm titanium alloy cortical screw allows firm fixation of the staple. A small deltopectoral or axillary approach is used and the subscapularis and capsule are incised on the line of the muscle fibres. The operation is relatively simple and combines the advantages of a Bankart, bone block and capsular plication in a single small operative procedure. Full movement, including external rotation, can be started 3 days postoperatively, and most patients can return to manual labour or contact sport in 4 to 6 weeks. A total of 47 anterior dislocations and two posterior dislocations have been treated since 1981, including six failed Putti-Platt operations and one Helfet operation. Full shoulder movement is usually achieved within 3 weeks of operation. PMID- 8406748 TI - Locked intramedullary nailing of humeral fractures. AB - Of 46 patients, 30 with fresh fractures of the humerus, nine with non-unions and seven with pathological fractures were treated with a new locked intramedullary nail. Of 30 patients with a fresh humeral fracture, three were lost to follow-up. All fresh fractures healed within 4 months. Functional results of the fresh fracture group were excellent in eighteen patients and satisfactory in three patients. Two patients with Neer type 6 fractures had unsatisfactory shoulder function; in the four other patients poor shoulder function resulted from a pre existing condition. Out of nine non-unions, six united within 6 months. The three other patients with atrophic non-union required bone-grafting later, after which consolidation was obtained. The long functional recovery period of the non-union group was related to the pre-existing limited shoulder function. The seven patients with a pathological fracture died within 8 months of operation. While alive they were free from pain and could be nursed well. PMID- 8406749 TI - Tibial fractures treated with the AO unreamed tibial nail. AB - During the last 2 years 20 fractures of the tibial shaft have been treated with the new, 'unreamed' solid AO tibial nail (UTN). There were 13 open fractures, while four of the seven closed fractures had severe soft tissue injuries. There were nine fractures classified as 'complex' or type C according to the AO classification. All UTNs were introduced without reaming but with proximal and distal interlocking. In all, 12 fractures (five of them open) were stabilized primarily by 'unreamed' nailing, whereas the first eight (open) fractures of the series were fixed initially by an external fixator and then nailed after about 14 days. In our series no intraoperative complications occurred, and no major soft tissue problems or infections were observed. The functional results were generally good; however, fracture healing appeared to be delayed in six cases. One patient had a further operation because of non-union. Intramedullary nailing, without reaming, seems to be a reliable and safe treatment for closed and open fractures with severe soft tissue injuries as an alternative to external fixation. The use of a thin, solid nail combined with interlocking allows the stabilization even of complex fractures. Contrary to initial assumptions, the UTN can be used as a definitive implant with no need for further stabilization in the majority of cases. PMID- 8406750 TI - Supracondylar fracture of the humerus: malrotation versus cubitus varus deformity. AB - A study correlating the degree of medial rotational deformity of the distal humerus and the degree of cubitus varus deformity secondary to supracondylar fracture was performed in 23 patients who underwent corrective supracondylar osteotomy. The mean age of the patients at the time of operation was 10.9 years (range 5-14 years). The time interval from injury to operation averaged 3.2 years (range 1-6 years). A medial rotational deformity occurred in 20 cases. The degree of medial rotational deformity (MRD) averaged 16.2 degrees (range 0-34 degrees). Mean carrying angles (CA) of the deformed and normal sides were -19.6 degrees and 6.5 degrees, respectively. Mean humero-elbow-wrist (HEW) angles, measured from radiographs, of the deformed and normal sides, were -18.8 degrees and 7.7 degrees, respectively. There was no correlation between the degree of MRD and the degree of varus deformity, using as a comparison either the CA or the HEW angle of the deformed elbow or their differences from the normal side. PMID- 8406751 TI - Fracture of the occipital condyle. PMID- 8406752 TI - Lymphocele of the thigh: a complication following tourniquet application in arthroscopy. PMID- 8406753 TI - Thyroid storm following chest trauma. PMID- 8406754 TI - Dislocation of the atlas and occiput associated with a widened superior mediastinum. PMID- 8406755 TI - Superficial radial nerve entrapment within a radial fracture in a child. PMID- 8406756 TI - Non-union of scapula spine fracture treated by bone graft and plate fixation. PMID- 8406757 TI - Regional audit of pedestrian accident care. AB - In all, 160 serious pedestrian accidents (ISS > 15 or death), were recorded during a 12-month prospective study of all trauma in a population of 3.2 million. Of these, 35 died at scene, 125 arrived at hospital alive and 68 (54 per cent) subsequently died. There were 35 (22 per cent) children, and 62 per cent (39) were more than 60 years of age. Prehospital care significantly delayed transfer to hospital. In the accident and emergency department, only 38 per cent of those unconscious had a cervical collar applied, and only 67 per cent were intubated. Of those transferred for neurosurgical care, 34 per cent were not intubated. The Median Injury Severity Score for each outcome group was similar between age groups. The Revised Trauma Score and APACHE II score showed significant differences between those who lived and died. TRISS analysis revealed that 32 per cent of deaths and 12 per cent of survivors were unexpected. ATLS treatment protocols should be instituted for prehospital care and in all accident and emergency departments (A&E). PMID- 8406758 TI - Morbidity and timing of colostomy closure in trauma patients. AB - The reported morbidity of colostomy closure in trauma patients varies from 5 to 27 per cent. Low morbidity rates are cited as a factor favouring colostomy creation and against expanded indications for primary repair in the treatment of colonic injuries. In order to assess the morbidity of colostomy closure, we reviewed all colonic injuries from 1979 to 1991 at our institutions. In all, 86 trauma patients who underwent colostomy creation and closure were identified. There were 82 men and four women with an age range of 16 to 74 years (mean 28.1 years). Of these, 95 per cent (N = 82) resulted from penetrating trauma. Of the patients, 63 per cent (N = 54) received end colostomies and 81 per cent (N = 70) of the patients had associated injuries. Of the patients, 38 per cent (N = 33) had a complication with their initial operation. There were no deaths after colostomy closure, but a total morbidity of 24.4 per cent (N = 21) was noted. There were 11 anastomotic complications (two of which required repeat laparotomy) and nine wound infections. The average length of stay was 10.4 days. Morbidity was concentrated in the group who had complications at their initial hospitalization. This was especially true if these patients underwent closure earlier than 3 months after injury. Conversely, if the first operation was uncomplicated, waiting longer than 3 months to perform colostomy closure did not improve results further. PMID- 8406759 TI - Should women attending fracture clinics be counselled about osteoporosis? AB - Osteoporotic fractures cause much morbidity and mortality and are a great burden on health care resources. It is well established that hormone replacement therapy (HRT) reduces the risk of osteoporotic fracture; it is also known to be worth initiating therapy even when one osteoporotic fracture has occurred in order to reduce the incidence of further fractures. In this study, detailed questionnaires assessing risk factors for osteoporosis were given to all women aged 25-60 years attending a daily fracture clinic over a 10-week period, and their radiographs were studied for signs of early trabecular bone loss, in order to determine whether at-risk patients could be identified easily. The findings were that 50 per cent of the premenopausal women had lifestyle-related risk factors about which they could be counselled. Using current guidelines, 90 per cent of the peri and post-menopausal group should be considered for HRT. Overall, only 7 per cent of patients who should be on HRT were receiving it. It was concluded that most of those at risk can be identified very simply in the fracture clinic. This aspect of fracture management is currently being neglected. PMID- 8406760 TI - Injury patterns in motorcycle road racers: experience on the Isle of Man 1989 1991. AB - We report the first prospective study on the incidence of motorcycle accidents of the world's foremost motorcycle road-racing venue, the Isle of Man. Between 1989 and 1991 there were six meetings during which over 2500 riders took part. Racing on essentially normal country and town roads, the motorcycles can reach speeds in excess of 306 km/h and the average lap record is now 198 km/h. During the study, 175 motorcyclists were injured and 16 were killed (9.1 per cent). The most common injury patterns were soft tissue injury (114 riders), head injury (52) and fractures/dislocations (188), of which one-third were open. The injuries sustained were more frequent and of a greater severity than recreational motorcycling. The casualty rate was 15 times, and the fatality rate 87 times the national figures. Of the 16 fatalities only seven reached hospital alive, despite the use of helicopter air ambulance, and 14 had serious head injuries. No fatality was thought to have been avoidable on review. There were no cervical spine injuries and only two major abdominal injuries. Of the injured riders, 93 (56 per cent) were admitted and there were 170 surgical procedures undertaken on 54 riders. PMID- 8406761 TI - Helicopter transfer of trauma patients: the Isle of Man experience. AB - A 10-year study of the helicopter transfer of motorcyclists injured during the International Motorcycle Tourist Trophy races (TT) and the amateur Manx Grand Prix (MGP) is reported. A total of 266 riders was transferred from the scene of the accident to hospital by air. The overall mortality rate for those riders airlifted was 3.38 per cent. The figures are compared with similar emergency helicopter services. PMID- 8406762 TI - Helicopter mountain rescue of patients with head injury and/or multiple injuries in southern Switzerland 1980-1990. AB - The aim of prehospital treatment of head injuries and severe multiple injuries is to prevent additional cerebral damage. When accidents occur in remote mountain areas, time is lost covering the distance to the nearest cabin or village where the rescue team can be called by telephone. Rapid transport of a trained physician to the patient can save precious time and allows prompt control of respiration and circulation at the scene of the accident. In a series of 57 rescue operations between 1980 and 1990, we used a helicopter staffed by an emergency physician and equipped with a winch. The hospital mortality rate was 12 percent (7 out of 57 patients). At 6 months after the accident, a normal neurological state was registered in 44 of the 55 survivors (88 per cent), whereas mild neurological deficiencies were noted in six patients (12 per cent). No persistent coma or vegetative state was seen. This experience suggests that fast rescue by a helicopter equipped with a winch and with an experienced emergency physician on board in an effective way of preventing secondary cerebral damage after accidents in remote mountain areas. PMID- 8406763 TI - Cervical collars: a potential risk to the head-injured patient. AB - Interface pressures beneath six types of collar in volunteers were measured. Under certain types of collar mean interface pressures of more than 10 mmHg were obtained, with several individual readings around 30 mmHg. By causing jugular venous obstruction a similar rise in intracranial pressure may be produced inadvertently. The mechanisms of action for this are discussed. Those collars with the least tendency to produce jugular venous obstruction were of the moulded variety which extend support to the occiput, mandible and shoulder girdle. The authors therefore recommend this type of cervical splintage device where injury of the cervical spine is suspected. This paper emphasizes the potential danger to head-injured patients when the other devices are applied too tightly, producing a 'venous tourniquet' effect. PMID- 8406764 TI - Application of dexamethasone in the treatment of acute spinal cord injury. AB - This communication evaluates the clinical efficacy of dexamethasone in acute spinal cord injury. The results of treatment in 290 patients given dexamethasone were compared with those of the control group of 330 patients not treated with corticosteroids. Patients with complete injuries and those with incomplete injuries showed greater improvement both quantitatively and qualitatively after treatment with dexamethasone than those without corticosteroids. The slightly increased risk of complications such as gastrointestinal bleeding and delayed wound healing was noted. It is recommended that corticosteroids should be used within the first hours after spinal cord injury. PMID- 8406765 TI - Midazolam sedation for the reduction of Colles' fractures. AB - The treatment of Colles' fractures in the elderly comprises a heavy workload for both accident and orthopaedic departments. The initial management has important clinical and financial implications for patient and hospital. The demand is variable and the ability to respond must also be flexible. The choice of anaesthetic technique is therefore most important. In our experience, intravenous sedation with midazolam (a water soluble benzodiazepine) has proven to be safe and effective in providing good conditions for anatomical reduction of Colles' fractures on an outpatient basis. PMID- 8406766 TI - Length of hospital stay and outcome after femoral neck fracture: a prospective study comparing the performance of two hospitals. AB - Length of hospital stay and outcome after femoral neck fracture were compared in a prospective study between two adjacent hospitals. In matched populations, mean length of stay was 30.8 days at Hospital X and 15.7 days at Hospital Y. Need for rehousing, age over 80 years and new nursing home placement prolonged length of stay at Hospital X, but not at Hospital Y. Hospital X had an orthopaedic rehabilitation ward and returned 88 per cent of patients to their own home, placing 9 per cent admitted from home in nursing homes. Hospital Y returned 76 per cent of patients to their own home and 19 per cent to nursing homes. The rapid discharge policy of Hospital Y saved significant resources within the acute hospital at the expense of returning significantly fewer patients to their own homes. PMID- 8406767 TI - Negative scintigraphy despite spinal fractures in the multiply injured. AB - Skeletal injuries may be overlooked in multiply-injured patients. Skeletal scintigraphy, a very sensitive indicator of skeletal injuries, may give complementary information. We therefore performed scintigraphic screening in 20 patients with multiple injuries. In all, 38 fractures were diagnosed by plain skeletal radiography, and were also visualized scintigraphically. Furthermore, scintigraphy revealed a fracture of the pelvis in one patient and a tibial plateau fracture in another. Scintigraphy was, however, negative in three patients; two with fractures of the transverse processes in the lumbar spine and one with a cervical spine fracture of the articular processes with dislocation. The use of scintigraphy as a screening method in patients with multiple injuries may be helpful in the diagnosis of skeletal injuries. However, fractures of the vertebral column were not always visualized, possibly because of technically inadequate examinations in injured patients unable to co-operate. PMID- 8406768 TI - Biomechanical factors in patient selection for radiography after head injury. AB - In order to assess the predictive value of certain biomechanical parameters for skull fracture after head injury, a prospective analysis was undertaken of a series of 5416 head-injured patients. In each case an assessment was made at presentation as to the velocity of impact and the physical properties of the impacting agent. The incidence of skull vault fracture was then calculated for injuries sustained at different velocities and for different types of contact. The incidence of fracture was also calculated with respect to the presence of post-traumatic amnesia (PTA). The incidences (95 per cent confidence intervals) of skull fracture at low, medium and high speeds were 0.17 (0.0427-0.433), 1.99 (1.47-2.63) and 10.2 (7.41-12.6) per cent respectively. Fractures tended to occur with greater frequency after impacts against broad hard surfaces or small objects rather than against broad soft objects. The percentage incidence of fracture in adults suffering more than 5 min of PTA was 17.93 (12.4-23.5) compared with 0.674 (0.372-1.1) in those without amnesia. Patient selection for skull radiography after an apparently minor head injury can be guided by an assessment of post traumatic amnesia and the biomechanics of the injury. PMID- 8406769 TI - External fixation in comminuted upper femoral fractures. AB - External fixation of comminuted upper femoral fractures has not been studied widely. The minority of such fractures which cannot be fixed internally due to clinical and anatomical problems are generally treated in traction. Early external fixators were not sufficiently robust to hold these fractures and pin site problems are more common in femoral fixation than in the tibia. A study was undertaken including all patients with comminuted upper femoral fractures who were too unwell or otherwise unsuitable for internal fixation. The long-term results were comparable, if not superior to traction, and patient comfort and mobilization were much improved. PMID- 8406770 TI - Early mobilization after operative repair of ruptured Achilles tendon. AB - The results of 19 patients in a pilot study in which early mobilization after operative repairs of ruptured Achilles tendons are presented. Serial ultrasound scanning of the repaired tendon showed satisfactory healing. Muscle function was analysed isokinetically and gait symmetry was examined using kinetic analysis. Normal strength of plantarflexion and dorsiflexion was achieved with almost normal range of ankle movement. All patients had normal gait kinetic parameters and symmetry. Normal activities were regained rapidly, and 15 out of 19 patients returned to their pre-injury status. There were no reruptures. PMID- 8406771 TI - Ladder injuries. AB - A series of 66 patients injured in falls from ladders was studied prospectively to assess the mode of fall, whether or not the fall was preventable, and the range of injuries sustained. Of the patients, 47 (71 per cent) fell because their ladder fell, and up to 90 per cent of these injuries were preventable. A wide spectrum of injuries was noted, and the importance of a thorough examination of such patients is emphasized. The need for a public education programme to prevent such injuries has also been highlighted. PMID- 8406772 TI - Single artery replantation of totally avulsed scalp. AB - The treatment of choice for the avulsed scalp is microvascular replantation. After replantation there are numerous arterial anastomoses, with venous congestion. Usually an effort to establish inflow through two arterial anastomoses is made. We elected to anastomose one artery and multiple veins in each of our patients. We present three consecutive successful replantations of total scalp avulsion, using only one arterial anastomosis, and multiple veins. Surprisingly, two of the patients regained their ability to elevate their eyebrows actively. PMID- 8406773 TI - We could not see the wood for the tree. PMID- 8406774 TI - Retained glass foreign bodies: confirmation and localization by computerized tomography and ultrasonography. PMID- 8406775 TI - Pure rotational displacement of the distal tibial epiphysis: a case report and a review of the literature. PMID- 8406776 TI - Dislocation of scaphoid and fractured capitate in a child. PMID- 8406777 TI - 'Stress' fracture of the scapula. PMID- 8406778 TI - Use of a metal detector and aluminium Sellotape to improve the accuracy of the windowing of casts. PMID- 8406779 TI - Early evidence from the Medicare fee schedule: the sky isn't falling. PMID- 8406780 TI - And the mother of all political battles has not even begun.... PMID- 8406781 TI - The future of Medicare Volume Performance Standards. AB - A policy of Medicare Volume Performance Standards (MVPS) was established to control the rate of growth in Medicare physician spending. The standards control spending growth by lowering fee updates when spending exceeds a preset standard, or increasing updates when spending is below the standard. Separate standards are established for surgical and nonsurgical services. This paper examines the policy's weaknesses and proposes refinements that would enhance the equity of MVPS rewards and penalties, and therefore, the policy's long-term viability. Specifically, we suggest that MVPS policy permit differentials in standards across areas when utilization rates vary geographically and for some types of services when volume growth differs significantly. PMID- 8406782 TI - Growth in Medicare inpatient physician charges per admission, 1986-1989. AB - Recently, there has been increasing interest in alternative Volume Performance Standards centered around the hospital medical staff. Using a national sample of Medicare claims, this paper analyzes growth in Medicare inpatient physician charges per admission between 1986 and 1989, by specialty, type of service, and type of hospital. Although growing more slowly than outpatient services, inpatient physician service volume and intensity growth is substantial. Inpatient physician services grew at an annual rate of 8.3%-31% of which was attributable to prices, 27% to case mix, and 41% to volume and intensity. Services provided by cardiologists, gastroenterologists, and other medical specialists demonstrate annual volume and intensity increases exceeding 13% annually. Consultations, hospital visits, and specialist services account for a third of all services provided during the admission and also demonstrate rapid annual rates of growth. PMID- 8406783 TI - Refining Medicare Volume Performance Standards. AB - This commentary focuses on the current workings of Medicare's Volume Performance Standards and possible alternatives that would make the program more effective in containing costs. The author presents his own perspectives as well as examining the options put forward in the two related papers by Holahan and Auckerman and Miller and Welch published in this issue of Inquiry. PMID- 8406784 TI - Assessing the validity of the geographic practice cost indexes. AB - Data on physician practice inputs were used to test the degree to which the geographic practice cost indexes (GPCIs) of the Medicare physician payment schedule reflect geographic variation in input prices. For purposes of this study, input quantity information was collected through the American Medical Association's Socioeconomic Monitoring System survey in 1990 and 1991. These data, along with practice expense information, were used to construct unit input prices. The GPCIs were correlated with input prices; however, "real" or GPCI adjusted prices varied significantly across locations. We conclude that the GPCIs are useful, but imperfect measures of geographic differences in physician practice input prices. PMID- 8406785 TI - Medicaid physician fees and use of physician and hospital services. AB - This paper examines how physician fees affect use of physician and hospital services under the Medicaid program. Using data from the 1987 National Medical Expenditure Survey (NMES), it examines how Medicaid physician fee levels affect beneficiaries' probability of using ambulatory physician services, the site at which ambulatory physician care is usually received, and how that site affects level of use and probability of hospital admission. The results indicate that low Medicaid fees hamper access to office-based physicians and encourage use of hospital outpatient departments and emergency rooms. They also indicate that having an office-based doctor as a usual source of ambulatory physician care is associated with a higher frequency of visits and a lower probability of having an inpatient hospitalization. PMID- 8406786 TI - Sources of health insurance for the self employed: does differential taxation make a difference? AB - Health insurance obtained through employment has enabled many workers to take advantage of the favorable tax treatment of employer contributions to these benefits. However, the full deductibility of such employer payments as a business expense and their exemption from income taxation are not available to self employed owners of unincorporated businesses. We identify the sources of health insurance for the self employed and examine how this disparity in tax treatment is associated with health insurance status. Analyses of data from the 1987 National Medical Expenditure Survey reveal that the differential taxation of the unincorporated self employed is associated with lower rates of employment-related coverage for themselves and their workers. PMID- 8406787 TI - The association of ownership and system affiliation with the financial performance of inpatient psychiatric hospitals. AB - Using a pooled, cross-sectional time-series analysis, this study examines the relationship of financial performance of short-term, inpatient psychiatric hospitals to ownership and system affiliation. After controlling for market variables, case mix, the number of services, and bed size, the results indicate that for-profit psychiatric hospitals had higher revenues per adjusted discharge, higher expenses per adjusted discharge, and higher profitability compared to not for-profit hospitals. System affiliated psychiatric facilities had lower revenues per adjusted discharge, lower expenses per adjusted discharge, and lower profitability than freestanding facilities. PMID- 8406788 TI - Payment mechanisms, nonprice incentives, and organizational innovation in health care. AB - Health care organizational structures support alternative combinations of payment and nonprice incentives for physicians, hospitals, and other providers. Changing perceptions among purchasers of the relative effectiveness of particular mechanisms lead to changes in the market shares held by particular organizational structures. This paper uses principal-agent and transactions cost economics to develop a conceptual matrix for understanding organizational innovation in the health care system. It contrasts the differing incentives in retrospective versus prospective payment mechanisms, arms-length review versus peer review mechanisms for controlling inappropriate utilization, and "free choice of physician" versus selective contracting mechanisms for structuring the basic insurer-provider relationship. PMID- 8406789 TI - Antibody-mediated enhancement of Legionella pneumophila-induced interleukin 1 activity. AB - The ability of antibody specific for Legionella pneumophila to enhance the induction of interleukin 1 (IL-1) production by murine peritoneal, splenic, and pulmonary macrophages in response to the bacterium was examined. Two preparations of L. pneumophila were utilized, a formalin-killed whole-cell preparation and viable bacteria. We measured both secreted (sIL-1) and cell membrane-associated (mIL-1) activities after incubation of the macrophages with the bacterial preparations in the presence or absence of the antibody. Both bacterial preparations induced sIL-1 and mIL-1 activities in each of the macrophage populations tested. These activities were generally enhanced by pretreating the bacteria with antibody, with the greatest enhancing activity observed for the formalin-killed preparations at lower doses of bacteria. PMID- 8406790 TI - Expression of the Haemophilus influenzae transferrin receptor is repressible by hemin but not elemental iron alone. AB - The absolute requirement for elemental iron and the porphyrin nucleus for growth of Haemophilus influenzae led us to investigate the role of iron and hemin in regulation of expression of the H. influenzae transferrin receptor. H. influenzae type b strain H1689 was grown in brain heart infusion broth supplemented with beta-NAD and either 10 or 0.1 microgram of hemin ml-1. Transferrin-binding ability was determined with a dot blot assay using human transferrin-horseradish peroxidase conjugate. Cells grown in media with 0.1 microgram of hemin ml-1 bound transferrin, but organisms grown in media with 10 micrograms ml-1 did not. In hemin-restricted media, transferrin binding occurred despite addition of up to 10 mM ferric nitrate, ferric citrate, or ferric PPi, whereas addition of 10 micrograms of hemoglobin ml-1 repressed expression. The breadth of species distribution of this mode of regulation was determined with strains previously characterized by multilocus enzyme electrophoresis. When grown in hemin restricted media, 24 of 28 type b strains and 52 of 57 serologically nontypeable strains exhibited transferrin binding, although none did so in hemin- and iron sufficient media. Strain H1689 and serologically nontypeable strain HI1423 grown in heat-inactivated pooled normal human serum, human cerebrospinal fluid, or human breast milk exhibited transferrin binding. Growth in these fluids with 10 micrograms of added hemin ml-1 abolished transferrin binding, whereas addition of 10 mM ferric nitrate did not. These data suggest that the transferrin receptor of H. influenzae is regulated by levels of hemin but not elemental iron alone and that this property is widely distributed among several major cloned families in the species. PMID- 8406791 TI - Cytokine mRNA in the central nervous system of SCID mice infected with Toxoplasma gondii: importance of T-cell-independent regulation of resistance to T. gondii. AB - Levels of cytokine mRNA were studied in the central nervous system (CNS) of SCID mice infected with Toxoplasma gondii. This infection led to 100% mortality by day 23 postinfection. Inflammation was observed in the lungs on day 7 and in the heart, liver, and kidneys on days 14 and 18 of infection. In the CNS, necrotic, acellular lesions that contained numerous parasites, accompanied by a localized astrocyte activation, were evident on day 14. Polymerase chain reaction-assisted amplification of RNA revealed that, although transcripts for interleukin-1 alpha (IL-1 alpha) and IL-1 beta were present in the brains of uninfected mice, increased levels of these transcripts were detected on day 7 of infection. Transcripts for macrophage inflammatory protein 1 and transforming growth factor beta were also detected in brains of infected mice at this time point. On days 14 and 18, levels of these transcripts had increased and transcripts for IL-6, IL 10, gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and granulocyte-macrophage colony-stimulating factor (GM-CSF) were also detected. Transcripts for IL-2 or IL-4 were not detected at any of the time points. Detection of locally produced cytokine transcripts may reflect involvement of the cytokines in the immunopathogenesis of this infection or involvement in mediating antitoxoplasma activity. To assess the possible role of endogenous IFN-gamma, TNF alpha, IL-10, IL-6, and GM-CSF, cytokine-neutralizing monoclonal antibodies were administered to infected SCID mice. Neutralization of IFN-gamma or TNF-alpha led to earlier mortality than that in controls. In contrast, treatment with antibody to IL-10 and IL-6 increased survival time. Treatment with anti-GM-CSF did not alter the time to death. These results indicate that TNF-alpha and IFN-gamma are both involved in T-cell-independent mechanisms of resistance to T. gondii in SCID mice and that IL-10 and IL-6 may downregulate the immune response to this pathogen. PMID- 8406792 TI - Adherence of Helicobacter pylori to primary human gastrointestinal cells. AB - Helicobacter pylori adheres only to gastric cells in vivo. However, the organism adheres to a wide variety of nongastric cells in vitro. In this study, we have used flow cytometry to assess the adherence of H. pylori to primary epithelial cells isolated from gastric, duodenal, and colonic biopsy specimens by collagenase digestion. After incubation of bacteria and cells together and subsequent staining with a two-stage fluorescein isothiocyanate-labelled H. pylori antibody method, cells with adherent bacteria could be easily distinguished from cells without bacteria. Binding to Kato III cells (a gastric adenocarcinoma cell line) was saturable when bacteria and cells were mixed at a ratio of 250:1. Adherence to cells isolated from gastric biopsy specimens was significantly better than adherence to cells isolated from duodenal or colonic biopsy specimens. Almost 70% of gastric cells had bacteria bound, in contrast to 30% of duodenal cells and 32% of colonic cells (P < 0.0001). There was no correlation between expression of hemagglutinins by the bacteria and ability to bind to either Kato III cells or primary epithelial cells isolated from gastric biopsy specimens. In view of the strict tropism that the organism exhibits in vivo for gastric cells, the results of this study indicate that primary cells are ideal for assessing the factors that might play a role in the pathogenesis of disease caused by the organism. PMID- 8406793 TI - Bordetella pertussis induces apoptosis in macrophages: role of adenylate cyclase hemolysin. AB - Bordetella pertussis, the causative agent of whooping cough, has been shown recently to enter and survive in epithelial cells and macrophages in vitro. In the present study, we show that B. pertussis is cytotoxic for J774A.1 cells, a monocyte-macrophage cell line, and for murine alveolar macrophages. We demonstrate that cell cytotoxicity mediated by B. pertussis occurred through apoptosis, as shown by changes in nuclear morphology and by host cell DNA fragmentation. Parental strains and a mutant deficient in pertussis toxin expression are able to induce apoptosis, whereas avirulent mutant or adenylate cyclase-hemolysin-deficient mutants are not cytotoxic. Both adenylate cyclase and hemolytic activities are required for programmed cell death. These results show that induction of apoptosis is dependent on the expression of adenylate cyclase hemolysin. The infection of murine alveolar macrophages in primary culture with B. pertussis leads to apoptosis, suggesting that this process might be relevant in vivo. The ability of B. pertussis to promote cell death may be important for the initiation of infection, bacterial survival, and escape of the host immune response. PMID- 8406794 TI - Virulence of Bordetella bronchiseptica: role of adenylate cyclase-hemolysin. AB - Bordetella bronchiseptica is a pathogen of laboratory, domestic, and wild animals and sometimes of humans. In the present study some characteristics of the virulence of B. bronchiseptica isolates of different origin were studied. All isolates had similar phenotypes, similar bacteriological characters, and synthesized adenylate cyclase-hemolysin, filamentous hemagglutinin and pertactin but not pertussis toxin. These isolates, however, differed in their ability to express dermonecrotic toxin and to cause a lethal infection, but no correlation was found with the human or animal origin of the isolates. The fact that the most virulent isolate did not express dermonecrotic toxin suggests that this toxin does not play an important role in the virulence of the bacteria in the murine model. After infection with virulent B. bronchiseptica a very early synthesis and a persistence of anti-adenylate cyclase-hemolysin and anti-filamentous hemagglutinin antibodies were observed in the sera of infected mice, suggesting a persistence of the bacteria or of its antigens. B. bronchiseptica adenylate cyclase-hemolysin was purified and was shown to be a major protective antigen against B. bronchiseptica infection. Furthermore, we showed that its immunological and protective properties were different from that of B. pertussis adenylate cyclase-hemolysin, confirming that Bordetella species are immunologically different. PMID- 8406795 TI - Anti-Cryptosporidium parvum antibodies inhibit infectivity in vitro and in vivo. AB - Cryptosporidium parvum causes acute diarrhea in immunocompetent individuals and a severe life-threatening disease in immunocompromised individuals, including AIDs patients. No efficacious therapy for cryptosporidiosis has yet been reported. However, treatment of some patients with cryptosporidiosis with hyperimmune bovine colostrum has ameliorated or eliminated clinical symptoms. Consequently, it is important to identify and characterize C. parvum antigens which are the targets of protective antibodies to facilitate the development of more efficacious therapy. We report that hyperimmune bovine colostral immunoglobulin inhibits C. parvum infectivity in a reproducible in vitro assay, and we correlate this inhibition with the protective capacity of the bovine colostrum in vivo. We have also identified the major C. parvum sporozoite antigens recognized on Western blots (immunoblots) by this colostral immunoglobulin preparation. Antibodies that recognize some surface molecules of other Apicomplexan parasites are protective in vivo. Consequently, we radioiodinated membrane proteins of sporozoites and immunoprecipitated 19 molecules which are the target of immunoglobulin that is protective in vivo and in vitro. PMID- 8406796 TI - Expression and characterization of the eaeA gene product of Escherichia coli serotype O157:H7. AB - In enteropathogenic Escherichia coli, the eaeA gene produces a 94-kDa outer membrane protein called intimin which has been shown to be necessary but not sufficient to produce the attaching-and-effacing lesion. The purpose of this study was to characterize the intimin specified by the eaeA allele of the enterohemorrhagic E. coli (EHEC) serotype O157:H7 strain CL8 and to determine its role in adherence. The carboxyl-terminal 266 amino acids of the CL8 intimin were expressed as a protein fusion with glutathione S-transferase, which was used to raise antiserum in rabbits. The antiserum reacted in Western immunoblots with a 97-kDa outer membrane protein of EHEC strains of serogroups O5, O26, O111, and O157 and enteropathogenic E. coli strains of serogroups O55 and O127. Surface labelling of CL8 with 125I showed that intimin was surface exposed. An eaeA insertional inactivation mutant of CL8 was produced and was designated CL8-KO1. Total adherence of CL8-KO1 to HEp-2 cells was not significantly different from that of CL8, but CL8-KO1 gave a negative result in the fluorescent actin staining test. The eaeA gene expressed alone in E. coli HB101 also gave a negative fluorescent actin staining test result. The eaeA gene of CL8 was able to complement the eaeA deletion mutation in CVD206. We conclude that the product of the EHEC eaeA gene is a 97-kDa surface-exposed protein and propose that it be designated intiminO157. Sherman et al. described a 94-kDa outer membrane protein which played an important role in adherence of E. coli O157:H7 (Infect. Immun. 59:890-899, 1991). Western immunoblotting and indirect fluorescent antibody studies showed that the protein described by Sherman et al. is not intimin. PMID- 8406797 TI - Expression of the major outer membrane protein of Chlamydia trachomatis in Escherichia coli. AB - The major outer membrane protein (MOMP) of Chlamydia trachomatis was expressed in Escherichia coli. To assess whether it assembled into a conformationally correct structure at the cell surface, we characterized the recombinant MOMP (rMOMP) by Western immunoblot analysis, indirect immunofluorescence, and immunoprecipitation with monoclonal antibodies (MAbs) that recognize contiguous and conformational MOMP epitopes. Western blot analysis showed that most of the rMOMP comigrated with authentic monomer MOMP, indicating that its signal peptide was recognized and cleaved by E. coli. The rMOMP could not be detected on the cell surface of viable or formalin-killed E. coli organisms by indirect immunofluorescence staining with a MAb specific for a MOMP contiguous epitope. In contrast, the same MAb readily stained rMOMP-expressing E. coli cells that had been permeabilized by methanol fixation. A MAb that recognizes a conformational MOMP epitope and reacted strongly with formalin- or methanol-fixed elementary bodies failed to stain formalin- or methanol-fixed E. coli expressing rMOMP. Moreover, this MAb did not immunoprecipitate rMOMP from expressing E. coli cells even though it precipitated the authentic protein from lysates of C. trachomatis elementary bodies. Therefore we concluded that rMOMP was not localized to the E. coli cell surface and was not recognizable by a conformation-dependent antibody. These results indicate that rMOMP expressed by E. coli is unlikely to serve as an accurate model of MOMP structure and function. They also question the utility of rMOMP as a source of immunogen for eliciting neutralizing antibodies against conformational antigenic sites of the protein. PMID- 8406798 TI - Isolation and characterization of a secreted metalloprotease of Aspergillus fumigatus. AB - A metalloprotease (MEP) secreted by Aspergillus fumigatus was isolated from an alkaline protease-deficient mutant after the fungus was cultivated in the presence of collagen as the sole nitrogen and carbon source. The enzyme was purified 50-fold from the culture supernatant after adsorption to hydroxylapatite and carboxy-methyl-Sephadex and after gel filtration. The molecular mass was determined to be 40 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point was estimated at pH 5.5 by isoelectric focusing. Reducing agents and divalent cations strongly inhibited enzyme activity, whereas nonionic detergents had no effect. A. fumigatus MEP was totally inhibited by EDTA, 1,10-phenanthroline, and phosphoramidon but not by inhibitors specific for serine, aspartate, and cysteine proteases. MEP is not able to cleave elastin and is thermosensitive. Sera from patients suffering from aspergilloma reacted with MEP in Western blotting (immunoblotting) analyses, suggesting that MEP promotes an antigenic response in these patients. PMID- 8406799 TI - Cytokine gene expression in human peripheral blood mononuclear cells stimulated by mannoprotein constituents from Candida albicans. AB - The expression of cytokine genes in cultures of human peripheral blood mononuclear cells (PBMC) stimulated with mannoprotein constituents (MP) of Candida albicans has been studied by means of S1 nuclease mapping analysis, polymerase chain reaction, and enzyme-linked immunosorbent assay. MP induced early, consistent, and long-lasting production of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha, and IL-6 mRNAs. Similar results were obtained when the same PBMC cultures were stimulated with the purified protein derivative (PPD) from Mycobacterium tuberculosis or with IL-2, although lower levels of IL-6 mRNA were detected in IL-2-stimulated cells than in MP- or PPD-stimulated cells. MP, PPD, and IL-2 induced appreciable levels of granulocyte-macrophage colony stimulating factor and gamma interferon, but only MP and PPD were able to induce IL-2 mRNA. MP were unable to stimulate a consistent expression of the genes encoding for IL-4, IL-5, and IL-10, while low, sometimes barely detectable levels of these cytokine mRNAs were observed in PPD- or IL-2-stimulated PBMC cultures. When protein synthesis of MP-stimulated PBMC was inhibited by cycloheximide, a superinduction of mRNAs for IL-4 and IL-10 and, more markedly, gamma interferon was observed. Overall, these results highlight the powerful, selective induction of cytokine gene expression by MP constituents of C. albicans in human PBMC cultures, thus providing some functional clues to explain the efficient state of the anticandidal response in normal human subjects. PMID- 8406800 TI - Collagen mediates adhesion of Streptococcus mutans to human dentin. AB - Some strains of Streptococcus mutans were found to recognize and bind collagen type I. Binding of 125I-labeled collagen type I was specific in that collagen types I and II, but not unrelated proteins, were able to inhibit binding of the labeled ligand to bacteria. Collagen binding to S. mutans was partially reversible and involved a limited number of bacterial binding sites per cell. S. mutans UA 140 cells bound collagen type I with high affinity (Kd = 8 x 10(-8) M). The number of binding sites per cell was 4 x 10(4). Collagen-binding strains of S. mutans were found to adhere to collagen-coated surfaces as well as to pulverized root tissue. S. mutans strains that did not bind the soluble ligand were unable to adhere to these substrata. Adherence to collagen-coated surfaces could be inhibited with collagen or clostridial collagenase-derived collagen peptides. Adherence of S. mutans to dentin was enhanced by collagen types I and II but inhibited by collagen peptides. S. mutans UA 140 bound significantly less 125I-collagen type I following treatment with peptidoglycan-degrading enzymes. These enzymes released a collagen-binding protein (collagen receptor) with a relative molecular size of 16 kDa. The results of this study suggest that collagen mediates adhesion of S. mutans to dentin. This interaction may target collagen-binding strains of S. mutans to dentin in the oral cavity and may play a role in the pathogenesis of root surface caries. PMID- 8406801 TI - Infection of murine macrophages with Toxoplasma gondii is associated with release of transforming growth factor beta and downregulation of expression of tumor necrosis factor receptors. AB - Toxoplasma gondii is capable of invading and multiplying within murine peritoneal macrophages. Previous studies have shown that treatment of macrophage monolayers with recombinant gamma interferon but not tumor necrosis factor (TNF) is associated with intracellular killing of T. gondii by macrophages. Furthermore, infection of macrophages with T. gondii prevents their stimulation for mycobactericidal activity by TNF. Since transforming growth factor beta (TGF beta) is known to suppress a number of functions in macrophages, we investigated the influence of infection with T. gondii on macrophage TNF receptors and on production of TGF-beta. Infection with T. gondii was associated with increased production of TGF-beta and downregulation of TNF receptors. This effect was observed early after infection and was partially inhibited by anti-TGF-beta 1 antibody. PMID- 8406802 TI - Growth in and breakdown of purified rabbit small intestinal mucin by Yersinia enterocolitica. AB - The mucus lining of the gastrointestinal tract serves as a protective barrier over the epithelial surface that must be crossed by invading bacteria seeking entry into the mucosa. The gel-forming component of mucus is mucin, a large polymeric glycoprotein. The present study examined the growth of Yersinia enterocolitica (with and without its virulence plasmid) in purified rabbit small intestinal mucin and the ability of bacteria to degrade mucin. Both virulent and nonvirulent organisms showed enhanced growth in mucin-supplemented media compared with unsupplemented media, but only at 37 degrees C and not at 25 degrees C. The effects of mucin were not specific because medium supplemented with bovine serum albumin also enhanced bacterial growth at 37 degrees C. Purified mucin was broken down into lower-molecular-weight components (assessed by monitoring its elution profile on a Sepharose CL-2B column) by plasmid-bearing Y. enterocolitica but not by plasmid-cured organisms. Culturing virulent Y. enterocolitica at 25 degrees C completely suppressed its capacity to degrade mucin, suggesting that this activity depends on plasmid expression. These results were confirmed in similar studies with purified rabbit colonic mucin. Mucin-degrading activity could be demonstrated in spent culture media from virulent Y. enterocolitica incubated at 37 degrees C but not in bacterial membrane preparations. Changes in the elution profiles of small intestinal and colonic mucins exposed to plasmid-bearing Y. enterocolitica at 37 degrees C were consistent with proteolytic depolymerization. The ability to grow well in mucin may help Y. enterocolitica to colonize the intestine, while the production of a mucin-degrading enzyme(s) by plasmid-bearing organisms may assist pathogenic strains to solubilize and penetrate the mucus gel layer. PMID- 8406803 TI - Characterization of the tpr gene product and isolation of a specific protease deficient mutant of Porphyromonas gingivalis W83. AB - The previously described protease gene (tpr) of Porphyromonas gingivalis W83 was shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the recombinant protein and in vitro translation to encode a 50-kDa protein whose active form migrates with an apparent molecular mass of 90 kDa. The 50-kDa protein was expressed at high levels by using a T7 RNA polymerase/promoter system. The NH2-terminal sequence of the protein was identical to the amino acid sequence deduced from the DNA sequence of the protease gene. Affinity-purified antibody to the 90-kDa recombinant protease reacted with an 80-kDa P. gingivalis protein. A specific protease (Tpr)-deficient isogenic mutant of P. gingivalis was generated by homologous recombination between P. gingivalis chromosomal DNA and a suicide plasmid carrying the cloned gene disrupted by insertion of an erythromycin resistance gene. Gelatin substrate zymography showed that cell extracts of the mutant lacked a protease band that migrated with an apparent molecular mass of 80 kDa. Western immunoblots of the cell extracts indicated the loss of an antigen with a similar mass. PMID- 8406804 TI - Transfer of antibody against Borrelia duttonii from mother to young in ddY mice. AB - The route of transfer of anti-Borrelia duttonii antibody subclasses from mother to young and their role in protection against borrelial challenge infection in ddY mice were investigated. Offspring from infected and noninfected mice were segregated and nursed by noninfected or infected mothers. Enzyme-linked immunosorbent assay analysis of antibodies of the cross-suckled offspring revealed that anti-B. duttonii immunoglobulin G1 (IgG1) is transferred exclusively in milk and that IgG2a is transferred mainly in milk but also slightly through the yolk sac route. On the other hand, IgG3 is transferred mainly through the yolk sac route but also slightly in milk, whereas IgG2b is transferred through both routes but to a lesser extent. Anti-borrelial IgM was not detected in any offspring. The protective role of transferred IgG subclasses was examined by challenge infection with B. duttonii. Offspring from noninfected mice fed by infected mothers had IgG1, IgG2a, and IgG3 at challenge and were completely protected against the challenge infection. On the other hand, offspring from infected mice fed by noninfected mothers had only IgG3, and 8 of 10 were completely protected from challenge infection whereas the other 2 contracted slight and transient spirochetemia. These findings suggested that anti borrelial IgG3 alone has considerable protective activity and that IgG1, IgG2a, or both, either by themselves or together with IgG3, have a complete protective activity against borrelial infection. PMID- 8406805 TI - Induction of release of tumor necrosis factor from human monocytes by staphylococci and staphylococcal peptidoglycans. AB - The role of cytokines in gram-positive infections is still relatively poorly defined. The purpose of this study was to establish whether or not intact staphylococci and purified peptidoglycans and peptidoglycan components derived from staphylococci are capable of stimulating the release of tumor necrosis factor (TNF) by human monocytes. We show here that intact staphylococci and purified peptidoglycans, isolated from three Staphylococcus epidermidis and three S. aureus strains, were indeed able to induce secretion of TNF by human monocytes in a concentration-dependent fashion. TNF release was detected by both enzyme immunoassay and the L929 fibroblast bioassay. In the enzyme immunoassay, a minimal concentration of peptidoglycan of 1 micrograms/ml was required to detect TNF release by monocytes, whereas in the bioassay a peptidoglycan concentration of 10 micrograms/ml was needed to detect a similar amount of TNF release. Peptidoglycan components such as the stem peptide, tetra- and pentaglycine, and muramyl dipeptide were unable to induce TNF release from human monocytes. PMID- 8406806 TI - Regulation of murine macrophage effector functions by lipoarabinomannan from mycobacterial strains with different degrees of virulence. AB - Lipoarabinomannan (LAM) is the major arabinose- and mannose-containing phosphorylated lipopolysaccharide (LPS) in mycobacterial cell walls. LAM preparations from a virulent strain (Erdman) (LAM(Erdman)) and an attenuated strain (H37Ra) (LAMH37Ra) of Mycobacterium tuberculosis, as well as from M. leprae (a virulent mycobacterium), were analyzed for their effects on various macrophage (M phi) effector functions. LAMH37Ra, like gram-negative LPS, exhibited a dose-dependent ability to induce tumor necrosis factor alpha (TNF alpha) production in normal M phi, and gamma interferon (IFN-gamma) priming of the M phi greatly augmented the levels of TNF-alpha. However, the effects of LAMH37Ra were unaffected by polymyxin B, which totally abrogated the effects of LPS. LAM(Erdman) and LAM from M. leprae, on the other hand, induced virtually no TNF-alpha production. Analysis of M phi mRNA by reverse transcription-polymerase chain reaction revealed that the levels of production. Analysis of M phi mRNA by reverse transcription-polymerase chain reaction revealed that the levels of TNF alpha mRNA induced by the various preparations correlated with the levels of TNF alpha protein detected. Interestingly, both LAMH37Ra and LAM(Erdman) could block subsequent IFN-gamma- and LPS-induced M phi activation, a previously reported measure of the potent ability of LAM to down-regulate M phi effector functions. Two lines of evidence suggested, however, that M phi cyclooxygenase products did not play a role in this down-regulation. LAMH37Ra and LPS could induce the production of NO2- in both normal and IFN-gamma-primed M phi, whereas LAM(Erdman) could stimulate NO2- production only in primed M phi. Both LAMH37Ra and LAM(Erdman) could substitute for LPS as a triggering signal for IFN-gamma-primed M phi in a toxoplasma killing assay. The triggering ability of LAM(Erdman), however, was abrogated by an anti-TNF-alpha antibody, suggesting that sufficient TNF-alpha production was stimulated by LAM(Erdman) to drive a M phi function relevant in host resistance. Thus, mycobacterial LAM is a potent regulator of M phi functions, a fact that may have important consequences in mycobacterial disease. PMID- 8406807 TI - Coxiella burnetii penetration into the reproductive system of male mice, promoting sexual transmission of infection. AB - Coxiella burnetii bacteria penetrate different host organs via the bloodstream. In infected mice, bacteremia was observed during the first week of infection: later, bacteria were cultured from the spleens, livers, testes, epididymes, prostate, and semen; bacteriuria developed beginning from the second week of infection. On day 21 of infection, degenerative changes with aggregated macrophages containing bacteria were observed in capillary blood vessels and the surrounding tissues of the adipose envelope of the epididymis. C. burnetii was shed to semen from the urogenital tract, probably from an abscess in the epididymis. There they were bound to the surface of spermatozoal cells, mainly to their heads, suggesting specific adhesion. Bacteria shed to semen were transmitted to female mice by sexual contact; this was demonstrated by the detection of antibodies against C. burnetii antigens in the blood of females and later by the cultivation of bacteria from the spleens, livers, and amniotic fluids of female mice. PMID- 8406808 TI - Characterization of a cDNA clone encoding the carboxy-terminal domain of a 90 kilodalton surface antigen of Trypanosoma cruzi metacyclic trypomastigotes. AB - We have cloned and sequenced a cDNA for a metacyclic trypomastigote-specific glycoprotein with a molecular mass of 90 kDa, termed MTS-gp90. By immunoblotting, antibodies to the MTS-gp90 recombinant protein reacted exclusively with a 90-kDa antigen of metacyclic trypomastigotes. The insert of the MTS-gp90 cDNA clone strongly hybridized with a single 3.0-kb mRNA of metacyclic forms, whereas the hybridization signal with epimastigote mRNA was weak and those with RNAs from other developmental stages were negative, indicating that transcription of the MTS-gp90 gene is developmentally regulated. A series of experiments showed that the MTS-gp90 gene is present in multiple copies in the Trypanosoma cruzi genome, arranged in a nontandem manner, and that there are at least 40 copies of the gene per haploid genome. Sequence analysis of recombinant MTS-gp90 revealed 40 to 60% identity at the amino acid level with members of a family of mammalian stage specific, 85-kDa surface antigens of T. cruzi. However, there are considerable differences in the amino acid compositions outside the homology region. PMID- 8406809 TI - Candida-specific Th1-type responsiveness in mice with experimental vaginal candidiasis. AB - The role of systemic cell-mediated immunity (CMI) as a host defense mechanism in the vagina is poorly understood. Using a murine pseudoestrus model of experimental vaginal candidiasis, we previously found that animals given a vaginal inoculum of viable Candida albicans blastoconidia acquired a persistent vaginal infection and developed Candida-specific delayed-type hypersensitivity (DTH) responses. The present study was designed to characterize the peripheral CMI reactivity generated from the vaginal infection in mice and to determine whether pseudoestrus is a prerequisite for the induction of peripheral CMI reactivity. Mice treated or not treated with estrogen and given a vaginal inoculum of C. albicans blastoconidia were examined for 4 weeks for their vaginal Candida burden and peripheral CMI reactivity, including DTH responsiveness and in vitro Th1 (interleukin-2 [IL-2], gamma interferon [IFN-gamma]/Th2 (IL-4, IL-10) type lymphokine production in response to Candida antigens. Results showed that although mice not treated with estrogen before being given a vaginal inoculum of C. albicans blastoconidia developed only a short-lived vaginal infection and harbored significantly fewer Candida CFU in the vagina compared with those given estrogen and then infected; DTH reactivity was equivalent in both groups. In vitro measurement of CMI reactivity further showed that lymph node cells from both estrogen- and non-estrogen-treated infected mice produced elevated levels of IL-2 and IFN-gamma in response to Candida antigens during the 4 weeks after vaginal inoculation. In contrast, lymph node cells from the same vaginally infected mice showed no IL-10 production and only small elevations of IL-4 during week 4 of infection. These results suggest that mice with experimental vaginal candidiasis develop predominantly Th1-type Candida-specific peripheral CMI reactivity and that similar patterns of Th1-type reactivity occur in mice regardless of the persistence of infection and the estrogen status of the infected mice. PMID- 8406810 TI - Modulation of adjuvant arthritis in Lewis rats by recombinant vaccinia virus expressing the human 60-kilodalton heat shock protein. AB - The immune response to the mycobacterial 65-kDa heat shock protein (hsp65) is considered an important event in the induction of adjuvant arthritis (AA) in rats; this induction probably occurs through a molecular mimicry mechanism involving cross-reactivity against the rat homolog hsp60. To analyze the role of mammalian molecule hsp60 in arthritis, we generated a recombinant vaccinia virus (hsp60-VV) carrying the human hsp60 gene inserted into the thymidine kinase locus under the control of the 7.5k vaccinia virus promoter. Human hsp60 is almost identical to its rat homolog (97.4% linear amino acid homology) and shares about 50% of amino acid positions with Mycobacterium tuberculosis hsp65. The latter supposedly carries a critical epitope for AA induction that is not present in human hsp60. Infections with hsp60-VV of monkey cell cultures led to the expression of the human hsp60 molecule, as evidenced by immunoblotting analysis with specific monoclonal antibodies. Also, Lewis rats infected with hsp60-VV produced specific antibodies, demonstrating the in vivo expression of human hsp60 in the infected animals. Therefore, we used hsp60-VV to analyze whether the delivery of hsp60 could affect the induction of AA in Lewis rats. hsp60-VV clearly reduced and retarded arthritic symptoms when administered to rats at day 7 after AA induction. In contrast, inoculation of rats with a control recombinant vaccinia virus did not affect the course of the disease. The improvement in AA with hsp60-VV administration was associated with a specific immune response, as determined by the presence of antibodies to hsp60 in the sera and the proliferation induced by hsp60 of T cells from popliteal lymph nodes. These results support a critical role for immunity to heat shock proteins in AA. Since the protective construct is virtually identical to rat homolog hsp60, we conclude that immunity directed to conserved areas of this family of proteins is directly involved in the pathogenesis of AA. PMID- 8406811 TI - Acid phosphatase activity in Coxiella burnetii: a possible virulence factor. AB - High-speed supernatant fluids derived from sonicated Coxiella burnetii contained considerable acid phosphatase activity when assayed by using 4 methylumbelliferylphosphate; they also contained a factor that blocked superoxide anion production by human neutrophils stimulated with formyl-Met-Leu-Phe. The pH optimum of the enzyme was approximately 5.0. The level of phosphatase activity detected in several isolates of C. burnetii implicated in acute (Nine Mile) and chronic (S Q217, PRS Q177, K Q154) Q fever was 25 to 60 times greater than that reported in other microorganisms, including Leishmania and Legionella spp. The enzyme was found in rickettsiae grown in different hosts (L929 cells and embryonated eggs) and, in the case of L929 cells, for both short periods (less than a month) and the long term (years). Cytochemical techniques coupled with electron microscopy localized the phosphatase activity to the periplasmic gap in the parasite. Ion-exchange chromatography revealed a major species of the enzyme and showed that the enzyme of the parasite was distinct from that of the host cell (L929 fibroblasts); its apparent molecular weight was 74,000. Phosphatase inhibitors (i.e., molybdate heteropolyanions) had differential effects on the phosphatases of the parasite and host cell. C. burnetii supernatant fluid inhibited superoxide anion production by formyl-Met-Leu-Phe-stimulated human neutrophils; molybdate inhibitors reversed the inhibition. Treatment of C. burnetii-infected L929 cells with one of the molybdate compounds (complex B') significantly reduced the level of infection and did not affect the viability or growth of the host cell. These data suggest that the acid phosphatase of the parasite may be a major virulence determinant, allowing the agent to avoid being killed during uptake by phagocytes and subsequently in the phagolysosome. PMID- 8406812 TI - Pertussis toxin-induced alterations of murine hepatic drug metabolism following administration of diphtheria and tetanus toxoids and pertussis vaccine adsorbed. AB - Administration of pertussis toxin (PT) in combination with diphtheria and tetanus toxoids adsorbed (DT vaccine) or with acellular pertussis vaccine adsorbed and diphtheria and tetanus toxoids (APDT) elicits dose- and time-dependent alterations in hepatic drug metabolism in mice. Cytochrome P-450 (P-450) levels were inhibited more than 50% at 7 days following a single injection of PT mixed with either vaccine. When combined with DT vaccine, 125 ng of PT was required to produce this effect, while as little as 16 ng of PT combined with APDT vaccine inhibited P-450 levels. The inhibition of P-450 levels is similar to that observed after a single injection of diphtheria and tetanus toxoids and pertussis vaccine adsorbed (DTP). Alterations of P-450 levels were accompanied by increased activities of quinone reductase but not with changes in plasma interleukin-6 or tumor necrosis factor levels. Other Bordetella pertussis virulence factors, such as filamentous hemagglutinin, fimbriae and pertactin, were also tested but had no significant effect on hepatic drug metabolism. Endotoxin or preparations containing endotoxin caused alterations in hepatic drug metabolism within 24 h, concomitant with increased interleukin-6 and tumor necrosis factor levels, but these effects had resolved by 1 week. DTP vaccine and preparations containing PT caused a marked induction of gamma interferon coincident with the maximal inhibition of P-450 levels. This effect was not present with DT or APDT vaccine alone, nor with endotoxin or any combination of factors that did not contain PT. These results demonstrate that PT is a necessary component for the sustained effects of DTP vaccine on hepatic drug metabolism and suggest a role for gamma interferon in this process. PMID- 8406813 TI - Role of gamma interferon in late stages of murine salmonellosis. AB - Gamma interferon (IFN-gamma) is an important mediator of natural resistance of mice to Salmonella species during the first week of infection, when it restricts the rate of intracellular growth of the bacteria but does not lead to their killing (A. Muotiala and P. H. Makela, Microb. Pathog. 8:135-141, 1990). We used the experimental mouse salmonellosis model to investigate the role of IFN-gamma in the later stages of a sublethal infection and the ensuing specific immunity. When anti-IFN-gamma was administered starting 6 days after challenge, it did not prevent the cessation of intracellular bacterial growth and the formation of the plateau stage in the second week of infection. In addition, anti-IFN-gamma given 14 and 16 days after challenge did not alter the elimination of the bacteria in the clearance stage in the third week of infection. When mice immunized 2 months previously with live vaccine were infected with virulent salmonellae, depletion of IFN-gamma enhanced the early growth of the bacteria in the same manner as that seen in naive mice. However, when the immunized mice were infected with attenuated aroA bacteria, their clearance started immediately and was unaffected by IFN-gamma depletion, demonstrating that IFN-gamma is not required for the clearance. We conclude that IFN-gamma restricts the rate of intracellular bacterial multiplication in the first week of Salmonella infection in both naive and immune mice but is not a mediator of bacterial clearance in either naive or immunized mice. PMID- 8406814 TI - Characterization of novel type C staphylococcal enterotoxins: biological and evolutionary implications. AB - The type C staphylococcal enterotoxins (SEC) are a group of highly conserved proteins with significant immunological cross-reactivity. Although three antigenically distinct SEC subtypes (SEC1, SEC2, and SEC3) have been reported in the literature, we observed that the isoelectric points of SEC from several Staphylococcus aureus isolates are different from those of any of these three subtypes. This observation led us to propose that additional SEC molecular variants exist. For assessment of this possibility, the sec genes from representative human, animal, and food isolates were cloned and sequenced. The toxins encoded by the 18 isolates used in this study included five unique SEC proteins in addition to SEC1, SEC2, and SEC3. Six of the SEC proteins (including SEC1, SEC2, and SEC3) were produced by human and food isolates. Analysis of seven bovine and ovine isolates showed that isolates from each animal species produced a unique host-specific SEC. All of the SEC caused lymphocyte proliferation, although some of the toxins differed in their ability to stimulate cells from several animal species. An explanation for these results, which is supported by our phenotypic analysis of Sec+ staphylococcal isolates, is that toxin heterogeneity is due to selection for modified SEC sequences that facilitate the survival of S. aureus isolates in their respective hosts. PMID- 8406815 TI - Glycolytic enzymes of Candida albicans are nonubiquitous immunogens during candidiasis. AB - A cDNA library was made with mRNA from Candida albicans grown under conditions favoring the hyphal form. The library was screened for sequences that encode immunogenic proteins by using pooled sera from five patients with oral candidiasis and five uninfected patients. Most of these patients were human immunodeficiency virus positive. From 40,000 cDNA clones screened, 83 positive clones were identified. Of these, 10 clones were chosen at random for further analysis. None of these 10 cDNAs were derived from a multigene family. The 5' and 3' ends of all 10 clones were analyzed by DNA sequencing. Two cDNAs were separate isolates of a sequence with strong homology to pyruvate kinase genes from other fungi (59 to 73%) and humans (60%). A third cDNA had strong sequence homology to the Saccharomyces cerevisiae and Kluyveromyces lactis alcohol dehydrogenase genes (68 to 73%). A fourth cDNA was homologous (81%) to an S. cerevisiae protein of unknown function. The functions of the remaining six C. albicans cDNAs are not known. A more detailed analysis of the clones encoding glycolytic enzymes revealed that sera from few patients recognized them as antigens. Therefore, although glycolytic enzymes constitute a group of C. albicans proteins that are immunogenic during oral and esophageal infections, their detection cannot be exploited as an accurate marker of infection. PMID- 8406816 TI - Optimizing oral vaccines: induction of systemic and mucosal B-cell and antibody responses to tetanus toxoid by use of cholera toxin as an adjuvant. AB - Cholera toxin (CT) is an effective mucosal antigen and acts as an adjuvant when given orally with various antigens; however, few studies have compared the levels of antibody responses to CT and coadministered protein in systemic and mucosal tissues. In this study, we used tetanus toxoid (TT) for assessment of immune responses. Time course and dose-response studies established that 250 micrograms of TT given orally with 10 micrograms of CT three times at weekly intervals induced high serum and gastrointestinal tract anti-TT and anti-CT antibody responses. Oral immunization with TT alone induced no detectable mucosal immunoglobulin A (IgA) antibodies in fecal extracts and only weak serum IgG anti TT responses. The coadministration of CT and TT induced peak serum IgG anti-TT responses following two oral doses that remained constant after the third oral immunization, while optimal mucosal IgA responses were seen after the third oral immunization. The serum anti-TT response obtained with CT and TT proved protective against TT challenge (100 minimum lethal doses), whereas mice orally given CT or TT alone died. Antigen-specific B-cell responses were assessed with an isotype-specific Elispot assay of isolated lymphoid cells from the spleen, Peyer's patches, and the small intestinal lamina propria. Interestingly, approximately fourfold-higher numbers of IgA anti-CT than of anti-TT antibody producing (spot-forming) cells occurred in lymphocytes from the lamina propria of mice orally immunized with both TT and CT. The adjuvant CT did not induce polyclonal B-cell responses in mice given CT by the oral route, since no significant differences in total numbers of B cells producing IgA, IgG, or IgM were found compared with the numbers in mice given TT alone. The results clearly indicate that serum and mucosal antibody responses develop with different kinetics and that protective TT-specific antibody responses are generated in the systemic compartment when TT is administered with CT via the oral route. PMID- 8406817 TI - Reproduction of porcine proliferative enteropathy with pure cultures of ileal symbiont intracellularis. AB - Porcine proliferative enteropathy is consistently associated with the presence of intracellular curved bacteria in epithelial cells in affected portions of intestine. Two strains of these intracellular bacteria were cultured in a cell culture system with rat enterocytes (IEC-18) and passaged several times and used as oral inocula for 14 gnotobiotic and 8 conventional pigs. DNA and immunological studies had identified these bacteria as belonging to a new taxon, Ileal symbiont (IS) intracellularis. Conventional pigs dosed with approximately 3.7 x 10(6) of these organisms passaged six times in cell culture developed severe lesions of proliferative enteropathy in the ileum. Other conventional pigs dosed with a lower titer or with organisms passaged 13 times developed moderate and minor lesions, respectively. All gnotobiotic pigs dosed with organisms failed to develop lesions. Control pigs, eight conventional and two gnotobiotic, dosed with diluent, uninfected cell material or left undosed failed to develop lesions also. Reisolation of IS intracellularis and demonstration of the organism in mucosal and fecal samples only occurred in conventional pigs dosed with organisms. Gnotobiotic pigs lacking a normal intestinal flora have not been shown to be colonized by the organism. Seroconversion to IS intracellularis or mucosal infiltration by inflammatory cells was not observed in experimentally affected pigs, confirming the weak immune response characteristic of the natural disease. These results support the identification of IS intracellularis as an etiological agent of proliferative enteropathy in pigs. PMID- 8406818 TI - Activation of the complement system in baboons challenged with live Escherichia coli: correlation with mortality and evidence for a biphasic activation pattern. AB - Activation of the complement system was studied in baboons that were challenged with live Escherichia coli. In the group challenged with a lethal dose (n = 4), the complement activation parameters C3b/c, C4b/c, and C5b-9 increased 13, 5, and 12 times the baseline value, respectively, during the first 6 h after the E. coli infusion, whereas in the group challenged with a sublethal dose (n = 10), they increased only moderately, by 2 to 3 times the baseline value. However, in this latter group, a more pronounced activation occurred at 24 h. Subsequent experiments showed that this second phase in complement activation started at 6 h after the challenge, at which time infused microorganisms had been cleared from the circulation. The simultaneous increase in C-reactive protein with this second phase suggested an endogenous activation mechanism involving this acute-phase protein. Levels of inactivated (modified) C1 inhibitor also increased in both groups, with peak levels of 2.5 times the baseline value at 24 h in the sublethal group and of 4 times at 6 h after the challenge in the lethal group. Thus, activation of complement in this animal model for sepsis occurs in a biphasic pattern, the initial phase mediated by the bacteria and the later phase mediated by an endogenous mechanism possibly involving C-reactive protein. The differences in complement activation between animals with lethal or sublethal sepsis support the hypothesis that complement activation contributes to the lethal complications of sepsis. PMID- 8406819 TI - Arcanobacterium haemolyticum phospholipase D is genetically and functionally similar to Corynebacterium pseudotuberculosis phospholipase D. AB - Arcanobacterium haemolyticum, a pathogen of the human upper respiratory tract and other systems, has been reported to produce soluble toxins, including a phospholipase D (PLD). We confirmed production of PLD by this organism and cloned and sequenced pld. Arcanobacterial PLD (PLD-A) was found to be a protein of approximately 31.5 kDa with a pI of approximately 9.4. Cosmid cloning, followed by subcloning into phagemid pBluescriptIISK+, yielded Escherichia coli(pAh140), a recombinant with a gene product corresponding to PLD-A. Evidence of PLD activity was found by three assays in supernatant fluid of cultures of E. coli(pAh140) and A. haemolyticum, but not in E. coli(pBluescriptIISK+). Experiments to determine if this protein was secreted were not conducted, but previous work with PLD from Corynebacterium pseudotuberculosis suggested that the presence of the enzyme in culture supernatant fluids was due to lysis of E. coli rather than to active transport. Antibodies in polyclonal sera from goats immunized with native or recombinant PLD-A neutralized native and recombinant PLD-A activity, and antibodies against native or recombinant PLD from C. pseudotuberculosis (PLD-P) partially neutralized native and recombinant PLD-A. Antibodies prepared against recombinant PLD-A labelled both recombinant and native PLD-A in Western blots (immunoblots) and dot blots, but antibodies against PLD-P did not. Sequencing of the insert in pAh140 revealed an open reading frame of 930 bp coding for 309 amino acids, including a putative signal sequence of 26 amino acids (3.2 kDa, determined on the basis of homology with the 24-amino-acid signal sequence of pld from C. pseudotuberculosis bv. ovis) and the mature PLD protein (31.5 kDa). Sequence comparisons of coding regions revealed 65% DNA homology with pld genes from C. pseudotuberculosis and Corynebacterium ulcerans. Comparison of amino acid sequences revealed 64% homology of PLD-A both with PLD-P and with PLD produced by C. ulcerans. PMID- 8406820 TI - Cation flux studies of the lesion induced in human erythrocyte membranes by the thermostable direct hemolysin of Vibrio parahaemolyticus. AB - Vibrio parahaemolyticus, an important agent of seafood-borne gastroenteritis, expresses several putative virulence factors that could account for the disease symptoms of infected humans, namely, diarrhea, nausea, and abdominal cramps. The pathogenicity of V. parahaemolyticus correlates well with the Kanagawa phenomenon (the hemolytic ability of strains grown on Wagatsuma blood agar), implicating the thermostable direct hemolysin (TDH) as the predominant toxin responsible for pathogenicity. TDH-induced hemolysis could be inhibited by the addition of the osmolyte sorbitol to the extracellular solution, supporting the hypothesis that hemolysis occurs through colloid osmosis secondary to an increase in the cation permeability of the membrane. The effect of TDH on cation permeability was investigated by measuring K+ (congener, 86Rb+) influx into human erythrocytes in which the endogenous cation transporters had been blocked (by use of ouabain, bumetanide, and nitrendipine). TDH increased K+ influx into these cells; this increase was rapid in onset and constant in magnitude, suggesting a direct action by TDH on the membrane. The kinetics of leak generation were examined; the relationship between counts accumulated and hematocrit indicated that the TDH induced lesion is multihit in nature. TDH-induced K+ influx was sensitive to Zn2+. Time courses of hemolysis in isosmotic solutions of monovalent cation chlorides were used to obtain the selectivity series for the TDH-induced leak: Cs+ > Li+ > K+ > Rb+ > Na+. Both the Zn2+ sensitivity and this selectivity series were obtained for crude culture supernatants, suggesting that TDH is the predominant leak-inducing agent. Thus, we have identified several features of the TDH-induced leak likely to be important in the diarrhetic action of V. parahaemolyticus in the human intestine. PMID- 8406821 TI - Tumor necrosis factor alpha, interleukin-1 alpha, interleukin-6, and prostaglandin E2 production in murine peritoneal macrophages infected with Ehrlichia risticii. AB - Ehrlichia risticii is a gram-negative obligate intracellular bacterium which primarily infects macrophages and crypt epithelial cells in the intestinal wall and is the etiologic agent of Potomac horse fever. To understand the pathogenesis of the disease, we tested whether E. risticii induces inflammation-associated products in thioglycolate-induced mouse peritoneal macrophages. Mouse peritoneal macrophages produced larger amounts of interleukin-1 alpha (IL-1 alpha) but lower levels of tumor necrosis factor alpha (TNF-alpha), IL-6, and prostaglandin E2 (PGE2) when exposed to live or killed E. risticii than when exposed to Escherichia coli lipopolysaccharide (LPS). Preincubation of macrophages with live or killed E. risticii suppressed TNF-alpha, IL-6, and PGE2 generation but not IL 1 alpha production in response to LPS. Murine gamma interferon treatment of macrophages did not influence TNF-alpha, IL-1 alpha, IL-6, or PGE2 production regardless of exposure to E. risticii. Intracellular cyclic AMP was significantly greater in E. risticii-infected macrophages than in uninfected macrophages. These results suggest that increased levels of IL-1 alpha but not TNF-alpha or PGE2 production by macrophages may be primarily involved in the pathogenesis of the disease caused by E. risticii. Increased intracellular concentration of cyclic AMP in infected macrophages may be chiefly responsible for the high level of IL-1 alpha and inhibition of TNF-alpha production in response to LPS. PMID- 8406822 TI - Inhibition of adherence of mucoid Pseudomonas aeruginosa by alginase, specific monoclonal antibodies, and antibiotics. AB - The adherence of pseudomonal species was investigated by using a newly developed radiometric dacron fiber microcolumn assay. Pseudomonas aeruginosa, P. stutzeri, and Xanthomonas maltophilia were more adherent (approximately 20%) than P. pseudomallei, P. fluorescens, and P. cepacia (approximately 10%). Mucoid strains of P. aeruginosa were consistently more adherent than nonmucoid strains (30% versus 20%). Alginase was shown to inhibit the adherence of mucoid but not nonmucoid P. aeruginosa. Monoclonal antibodies to alginate were also shown to inhibit the adherence of mucoid but not nonmucoid P. aeruginosa. In addition, antibiotics active against P. aeruginosa were shown to inhibit the adherence of both mucoid and nonmucoid strains. Furthermore, synergism between dyadic combinations of monoclonal antibodies and antibiotic (ciprofloxacin), as well as alginase and antibiotic, was also observed. These results indicate that bacterial alginate has an intrinsic role in the adherence of mucoid P. aeruginosa and may have evolved not only for protection against dehydration in the water and soil ecosystem of this bacterium, but also as a means of attaching to soil substrates in the same ecosystem to enhance survival. They also suggest that synergistic combinations of antibiotics with alginase or monoclonal antibodies to alginate may be of value in the therapy of some pseudomonal infections. PMID- 8406823 TI - Saliva-binding region of Streptococcus mutans surface protein antigen. AB - A 190-kDa surface protein antigen (PAc) of Streptococcus mutans binds to human salivary components. For detection of specific binding of the PAc protein to human salivary components, a simple sandwich assay was used. Microtiter plates precoated with recombinant PAc (rPAc), PAc fragments, or S. mutans whole cells were allowed to react with human whole saliva and then were incubated with biotinylated rPAc. The biotinylated rPAc bound to salivary components was detected by use of alkaline phosphatase-conjugated streptavidin and p nitrophenylphosphate. In this assay, the binding of whole cells of S. mutans and purified rPAc to salivary components was confirmed. For determination of a saliva binding region of the PAc molecule, 14 truncated PAc fragments were constructed by use of the polymerase chain reaction and an expression vector, pAX4a+. The binding of these truncated PAc fragments to human salivary components was determined by the sandwich assay. Among the truncated PAc fragments, fragments corresponding to residues 39 to 864 and residues 39 to 1000 of PAc showed a high ability to bind to salivary components. Shorter recombinant fragments corresponding to residues 39 to 217, residues 200 to 481, residues 470 to 749, and residues 688 to 864 did not exhibit any binding ability. The fragment that corresponds to a proline-rich repeating region (residues 828 to 1000) bound directly to the PAc protein. These results suggest that residues 39 864 of the PAc molecule are important in the binding of the surface protein to human salivary components, and the proline-rich repeating region of the PAc protein may contribute to spontaneous self-aggregation of the PAc protein. PMID- 8406824 TI - Chemical properties of lipopolysaccharides from spotted fever group rickettsiae and their common antigenicity with lipopolysaccharides from Proteus species. AB - The lipopolysaccharides (LPS) isolated from spotted fever group (SFG) rickettsia strains Thai tick typhus TT-118 and Katayama were characterized by chemical analyses, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, enzyme linked immunosorbent assay (ELISA), and immunoblotting. These LPS did not contain heptose, but they contained 3-deoxy-D-manno-octulosonic acid (KDO), glucosamine, quinovosamine, phosphate, ribose, an unknown neutral sugar, and palmitic acid. Resolution of the apparent molecular masses of these LPS by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and staining with silver showed ladder like bands. In an ELISA, convalescent-phase sera from 10 patients with Japanese spotted fever reacted with LPS from the Katayama strain, and 90% (9 of 10) of these sera also reacted with LPS isolated from Proteus vulgaris OX2. Immunoblotting revealed that the sera reacted with the high-molecular-mass bands of LPS from SFG rickettsiae, in addition to those of OX2 LPS. In an ELISA, immunoglobulin M antibodies from these sera reacted with the O-polysaccharide and lipid A portions of LPS from P. vulgaris OX2. The epitopes common to LPS of SFG rickettsiae and P. vulgaris OX2 may be in the O-polysaccharide and lipid A portions. PMID- 8406825 TI - Differential regulation of cytokine production in lipopolysaccharide tolerance in mice. AB - We investigated the pattern of down-regulation of cytokine production in endotoxin (lipopolysaccharide [LPS]) tolerance. A 4-day treatment with LPS (35 micrograms per mouse) was followed by a challenge on day 6 with one more injection of LPS. Circulating tumor necrosis factor (TNF) and interleukin-6 (IL 6) could not be induced (> 99% inhibition) by LPS in LPS-tolerant mice; colony stimulating factor (CSF) was also down-regulated by more than 95%, whereas interferon (IFN) and IL-1 syntheses were only partially inhibited. To study the mechanism of cytokine down-regulation in tolerance, we attempted to reverse the tolerant state by pretreatment with phorbol 12-myristate 13-acetate (PMA) (4 micrograms per mouse) 10 min before the LPS challenge. PMA completely restored IL 6 production and partially that of CSF. PMA had no effect on IFN production and inhibited the induction of IL-1. TNF production was also not restored by PMA. To investigate the role of endogenously produced cytokines in the development of LPS tolerance, we administered IL-6, TNF, or IL-1 alpha, using the same treatment schedule as that for LPS. Whereas IL-6 had no effect, IL-1 alpha or TNF induced partial tolerance to LPS in terms of inhibition of LPS-stimulated TNF and IL-6 production. However, a full LPS-tolerant state could not be induced by administration of recombinant cytokines, suggesting the existence of additional mechanisms, such as a loss of LPS receptors or changes in release of soluble binding proteins. PMID- 8406826 TI - Accelerated decay of C3b to iC3b when C3b is bound to the Cryptococcus neoformans capsule. AB - Incubation of encapsulated and nonencapsulated Cryptococcus neoformans in normal human serum (NHS) leads to activation and binding of potentially opsonic fragments of complement component C3 to the yeast cells. Analysis of the molecular forms of C3 after incubation of encapsulated cryptococci in NHS showed that the percentage of bound C3 occurring as iC3b approached 100% after 8 min. The percentage of bound C3 occurring as iC3b on nonencapsulated cryptococci never exceeded 70%, even after 60 min of incubation in NHS. Conversion of C3b to iC3b was assessed further by incubating C3b-coated cryptococci for various times with a mixture of complement factors H and I at 40% of their respective physiological concentrations. Most, if not all, of the C3b on encapsulated cryptococci was converted to iC3b at a single fast rate. Conversion of C3b to iC3b on nonencapsulated cryptococci did not follow a single rate constant and appeared to have a fast and a slow component. Studies of the requirements for factors H and I in cleavage of C3b to iC3b showed steep dose-response curves for both factors in the case of encapsulated cryptococci and shallow curves with C3b bound to nonencapsulated cryptococci. Taken together, our results indicate that C3b molecules bound to encapsulated cryptococci have a uniformly high susceptibility to conversion to iC3b by factors H and I. In contrast, a significant portion of the C3b bound to nonencapsulated cryptococci is very resistant to conversion to iC3b by factors H and I. PMID- 8406827 TI - Seroprotective groups among isolates of Borrelia burgdorferi. AB - We demonstrated that different seroprotective groups exist among isolates of Borrelia burgdorferi and Borrelia garinii. The major group was composed of isolates 297, B31, S-1-10, MMTI, IPT, and ATCC 35211 and 21 isolates obtained from California, Illinois, New York, Texas, and Wisconsin. A second group was composed of European isolates PBi and G25. A third group was composed of a single isolate, C-1-11. These groupings were supported by Western immunoblot findings. In addition, the seroprotective groups were confirmed by passive transfer of immune sera and challenge of recipient hamsters with the homologous isolate or other isolates of B. burgdorferi or B. garinii. These studies demonstrate that a monovalent vaccine will not provide complete protection against infection with all isolates of B. burgdorferi. PMID- 8406828 TI - Expression and secretion of an Arthrobacter dextranase in the oral bacterium Streptococcus gordonii. AB - We have constructed a plasmid to express and secrete dextranase in the oral bacterium Streptococcus gordonii. The dextranase gene from Arthrobacter sp. strain CB-8 was linked to a promoter and a DNA sequence encoding the signal peptide of Streptococcus downei glucosyltransferase I (gtfI) followed by the Escherichia coli rrnBt1t2 terminator and inserted in the shuttle vector pVA838. S. gordonii transformed with this plasmid (pMNK-4) expressed and secreted mature Arthrobacter dextranase. The transformant was found to repress the firm adherence of water-insoluble glucan in a coculture experiment with cariogenic bacteria, Streptococcus sobrinus, in the presence of sucrose. Such genetically engineered oral bacteria could provide a therapy to prevent dental caries. PMID- 8406829 TI - Molecular epidemiology of penicillin-resistant pneumococci isolated in Nairobi, Kenya. AB - A total of 26% of the pneumococci isolated from an outpatient clinic in Nairobi, Kenya, during 1991 to 1992 had intermediate levels of penicillin resistance. Gene fingerprinting and DNA sequencing were used to distinguish the penicillin-binding protein (PBP) 1A, 2B, and 2X genes in 23 resistant isolates. Isolates were grouped into those that had identical forms of each of the three PBP genes (fingerprint groups) and those that had identical rRNA gene restriction patterns (ribotypes). Both methods divided the isolates into 11 groups. In a few cases, horizontal gene transfer appeared to have distributed an identical altered PBP gene into different pneumococcal lineages. Eight isolates were indistinguishable by ribotyping or multilocus enzyme electrophoresis and contained identical PBP 1A genes. Although these isolates were therefore members of the same clone, they were divided into two fingerprint groups which contained different PBP 2X and 2B genes. Presumably, members of this clone have acquired different altered PBP 2X and 2B genes on two separate occasions. One of these fingerprint groups contained isolates of serotype 14, whereas the other contained isolates of both serotypes 14 and 7. The identification of isolates in the latter group that are identical by all criteria, except serotype, implies the occurrence of a change in serotype. The predominant serotypes of the penicillin-resistant pneumococci from Nairobi were serotypes 14 and 19. In both cases, isolates of the same serotype which required the same MIC of penicillin were not members of a single clone, indicating that identity of serotype and MIC are not sufficient criteria for defining clones of resistant pneumococci even when the bacteria are isolated from a single clinic. PMID- 8406830 TI - Identification of hydrogen peroxide as a Streptococcus pneumoniae toxin for rat alveolar epithelial cells. AB - Streptococcus pneumoniae infections of the lung are associated with significant damage to the alveolar epithelium. Host phagocytes and pneumolysin, a cytolytic toxin of S. pneumoniae, are believed to contribute to this cellular damage, yet experiments in which these elements are absent demonstrate the presence of an additional soluble S. pneumoniae factor that is toxic to alveolar epithelium. We examined the effects of S. pneumoniae-associated alveolar epithelial cell injury by factors other than S. pneumoniae-derived pneumolysin or phagocyte products by exposing cultured rat type II alveolar epithelial cells (RAEC) to S. pneumoniae mutants that lacked pneumolysin activity. We found that mutant pneumolysin deficient strains of S. pneumoniae produced injury to RAEC similar to that produced by the parent strains. A toxin of type 14 S. pneumoniae was distinguished from pneumolysin by physiochemical (i.e., molecular mass and heat stability) and functional (i.e., hemolytic activity and cytotoxic activity) properties and was identified as hydrogen peroxide. All S. pneumoniae strains tested produced hydrogen peroxide, and in many strains hydrogen peroxide production was comparable to that of activated neutrophils. We conclude that S. pneumoniae produces hydrogen peroxide in concentrations that are cytotoxic to RAEC in vitro and that alveolar epithelial damage due to hydrogen peroxide may be involved in the pathogenesis of host cellular injury in pneumococcal pneumonia. PMID- 8406831 TI - Isolation and preliminary characterization of the 14- to 18-kilodalton Candida albicans antigen as a phospholipomannan containing beta-1,2-linked oligomannosides. AB - Western blot (immunoblot) analysis of Candida albicans germ tube extracts has demonstrated the probable presence of beta-1,2-linked oligomannosides acting as epitopes distributed over a 14- to 18-kDa antigen unreactive to concanavalin A. These conclusions about the existence of these non-mannan-associated oligomannoside species were reinforced in the present study by the demonstration of reactivity of factor serum 5 (Iatron Laboratories) with the same antigen. A monoclonal antibody which reacted in an enzyme immunoassay with beta-1,2-linked oligomannosides converted into neoglycolipids and in Western blotting with the 14 to 18-kDa antigen from yeast and germ tubes, through metaperiodate-sensitive epitopes, was used for further characterization of the molecule. Reducing agents and strong protease digestion, which have deleterious effects on C. albicans proteins and mannoproteins, affected neither the antigenicity nor the relative molecular weight of the molecule. Western blots performed after migration of protease-treated extracts in polyacrylamide gels without sodium dodecyl sulfate (SDS) showed that the 14- to 18-kDa antigen could be negatively charged, whereas metabolic radiolabeling demonstrated that these charges could originate, at least in part, from the presence of phosphorus within the molecule. Chloroform-methanol water extraction of protease-resistant material led to purification of the 14- to 18-kDa antigen, as determined by SDS-polyacrylamide gel electrophoresis and Western blotting. Metabolic radiolabeling with mannose confirmed the presence of these sugar residues within the purified 14- to 18-kDa antigen (despite its nonreactivity to concanavalin A), whereas radiolabeling with palmitic acid demonstrated its lipopolysaccharidic nature. Together, these results led to the conclusion that the 14- to 18-kDa antigen is a phospholipomannan. PMID- 8406832 TI - A role for Bacteroides fragilis neuraminidase in bacterial growth in two model systems. AB - Two Bacteroides fragilis neuraminidase-deficient mutants were used to study the role of neuraminidase activity in growth of B. fragilis in tissue culture monolayers (CHO cells) and in the in vivo rat granuloma pouch. The nanH structural gene for neuraminidase was cloned from B. fragilis TM4000 and was used to create two isogenic strains with chromosomal disruptions at the nanH gene. B. fragilis VRC404 contains an insertion flanked by disrupted copies of the nanH gene, and B. fragilis VRC426 contains a deletion of a significant portion of nanH coding sequences. The insertion mutant VRC404 is capable of reverting to nanH+. It grew as well as the wild type in CHO monolayers. However, between 48 and 72 h after infection, the bacterial population was enriched with nanH+ bacterial cells (10 to 20%). In the rat pouch 48 h after infection, more than 90% of the population sampled had become nanH+. The deletion mutant VRC426 showed a severe growth defect in the rat pouch model. In addition, VRC426 was efficiently outgrown by the wild type in competition experiments, even when the mutant was present at 10 times the number of wild-type cells at the time of infection. A common characteristic of both model systems is a drastic decrease in the free glucose concentration 16 to 24 h postinfection. We suggest that neuraminidase activity may be required for B. fragilis to grow to maximal levels in the tissue culture and rat pouch systems by making other carbon sources available after glucose levels are reduced. PMID- 8406833 TI - Shigella flexneri invasion of HeLa cells induces NF-kappa B DNA-binding activity. AB - Although information about the genetic basis and mechanisms of Shigella flexneri cellular invasion is accumulating, little is known about changes in cell signaling and their consequences following bacterium-host cell interactions. A general result of signal transduction is alterations in the levels and/or activities of transcription factors. Alterations in transcription factor binding activities were observed following challenge with S. flexneri. Changes in the DNA binding activities of cellular transcription factors to AP1, AP2, cyclic AMP response element, CTF1/NF1, NF-kappa B/Rel, OCT1, and SP1 DNA-binding sites were investigated by electrophoretic mobility shift assays. NF-kappa B/Rel DNA-binding activity was enhanced more than 11-fold by cellular invasion; noninvasive S. flexneri strains induced low levels of kappa B DNA binding. Both subunits of the NF-kappa B transcription factor, p50 and p65, but not c-Rel (p85), are components of the kappa B DNA-binding activity. These data suggest that changes in cellular transcription factor binding activity are a consequence of S. flexneri invasion, and these changes could play a role in the initial host response or in the pathogenesis of the disease. PMID- 8406834 TI - PRB3 null mutations result in absence of the proline-rich glycoprotein Gl and abolish Fusobacterium nucleatum interactions with saliva in vitro. AB - The glycosylated proline-rich glycoprotein (Gl or PRG), a product of the PRB3 gene, is a major constituent of human parotid saliva. Important functions proposed for Gl include acting as a bacterial receptor. The Gl proteins of several subjects were typed by two polyacrylamide gel electrophoresis (PAGE) systems: acid-lactate PAGE followed by staining with the periodic acid-Schiff reagent and sodium dodecyl sulfate-PAGE followed by electrophoretic transfer and staining with amido black or concanavalin A. The results showed one subject who apparently lacked Gl. The four exons, including splice junctions, for both PRB3 alleles of this subject were completely sequenced. Unexpressed (null) mutations were detected with an identical C nucleotide insertion in the same coding region of exon 3 of both alleles. This C nucleotide insertion leads to a frameshift with a premature termination codon that probably results in markedly reduced or absent PRB3 gene expression. We then used a nitrocellulose blot overlay assay to assay the bacterial receptor activity of parotid saliva from the PRB3null subject. No interactions with Fusobacterium nucleatum, shown previously to interact selectively with Gl, were detected. Together, these results suggest that this subject does not express the PRB3 gene and that one of the consequences is an altered ability to interact with a bacterium known to colonize the oral cavity. PMID- 8406835 TI - Specific serum and local antibody responses against Cryptosporidium parvum during medication of calves with halofuginone lactate. AB - Fecal and serum anti-Cryptosporidium parvum immunoglobulin A (IgA), IgM, and IgG were monitored by an enzyme immunoassay in C. parvum-infected calves after medication with halofuginone lactate. In a first experiment, four groups of five 1-day-old colostrum-fed calves were inoculated with 10(6) oocysts of C. parvum. They were medicated with 0, 30, 60, or 120 micrograms of halofuginone lactate per kg from days 2 to 8 postinfection (p.i.). Unmedicated calves passed large numbers of oocysts between 3 and 14 days p.i. Treatment with 30 micrograms/kg did not completely inhibit oocyst output during medication, whereas 60 and 120 micrograms/kg did. The latter groups passed only a reduced number of oocysts when the drug was withdrawn. In a second experiment, 3- to 6-day-old colostrum-fed calves were divided into three groups of 16 or 17 animals each. All animals had acquired C. parvum infection before arrival at the fattening unit. They were medicated with 0, 60, or 120 micrograms/kg for 7 days beginning on the day of arrival. Unmedicated calves passed large numbers of oocysts from 0 to 21 days. Medication stopped oocyst output at day 7, but some of the calves again passed low numbers of oocysts 7 days after withdrawal of the drug. Experimental infection of unmedicated calves was followed by a rise in local anti-C. parvum IgA and IgM titers. Rising coproantibody levels coincided with falling oocyst output. In halofuginone-medicated and experimentally infected calves, only specific anti-C. parvum IgM levels rose during the first 5 days p.i. Specific IgA levels increased in association with oocyst output after withdrawal of the drug in the 60- and 120-micrograms/kg groups. In naturally infected calves, on the other hand, both specific IgA and IgM levels rose further during medication. Although titers were lower than in unmedicated controls, no significant differences were observed. Both medicated and unmedicated calves were equally protected from a challenge with 10(7) oocysts 16 weeks after the first contact with the parasite. PMID- 8406836 TI - The gene encoding phosphoribosylaminoimidazole carboxylase (ADE2) is essential for growth of Cryptococcus neoformans in cerebrospinal fluid. AB - A cryptococcal meningitis model in corticosteroid-treated rabbits was used to assess the requirement for the phosphoribosylaminoimidazole gene (ADE2) for virulence of Cryptococcus neoformans. A wild-type strain (H99), an ade2 auxotroph of H99 (M001), and a randomly selected prototrophic transformant of M001 (M001.1c) which had received the cloned ADE2 cDNA copy were inoculated intrathecally into immunosuppressed rabbits. While M001 was avirulent in the central nervous system model, virulence was completely restored to wild-type pathogenicity in the prototrophic transformant. This study identifies the pathogenic importance of an endogenous adenine pathway in this yeast and confirms that purine biosynthesis is a potential target for antifungal therapy. It also demonstrates that the virulence of C. neoformans can be molecularly changed and detected within a clinically relevant animal model. PMID- 8406837 TI - CVD110, an attenuated Vibrio cholerae O1 El Tor live oral vaccine strain. AB - The recent expansion of the seventh cholera pandemic into South America emphasizes the need for a safe, long-lasting, protective, and nonreactogenic vaccine for this disease. Since the predominant Vibrio cholerae O1 strains in the world today are of the El Tor biotype, a bivalent vaccine containing both classical and El Tor biotypes may be desirable. We have constructed a new oral vaccine candidate, V. cholerae CVD110 El Tor, Ogawa, from which all toxin genes so far identified in V. cholerae have been deleted. Three of these genes, those encoding cholera toxin (ctx), zonula occludens toxin (zot), and accessory cholera enterotoxin (ace), are located on a 4.5-kb virulence cassette flanked by repetitive sequences (RS1 elements). Homologous recombination between these RS1 elements resulted in the deletion of this virulence cassette to yield V. cholerae CVD109. Insertion of genes encoding mercury resistance (mer) and the cholera toxin B subunit (ctxB) into the hemolysin locus (hlyA) produced CVD110. This insertion serves three purpose. (i) It genetically tags the vaccine strain so as to distinguish it from wild-type V. cholerae O1. (ii) It produces cholera toxin B subunit in order to elicit antitoxic immunity. (iii) It inactivates the hemolysin gene, rendering the strain nonhemolytic on sheep erythrocyte plates. Supernatants from V. cholerae CVD110 cultures are nonreactogenic when assayed in Ussing chambers. PMID- 8406838 TI - The RESA-2 gene of Plasmodium falciparum is transcribed in several independent isolates. AB - The relevance of the ring-infected erythrocyte surface antigen (RESA) of Plasmodium falciparum as a malaria vaccine candidate has been questioned of late because RESA-deficient parasites have been found to multiply normally in culture or in monkeys. The RESA-2 gene was recently described as a pseudogene highly homologous to RESA. In this report, we demonstrate that RESA-2 is not a pseudogene, because we were able to detect RESA-2 transcripts in asexual blood stages of multiple isolates by using polymerase chain reaction on reverse transcribed mRNA. Transcription of RESA-2 was observed whether or not the isolates studied expressed the RESA protein. PMID- 8406839 TI - Regulation of slime production in Staphylococcus epidermidis by iron limitation. AB - Slime production by most strains of Staphylococcus epidermidis was enhanced by conditions of iron limitation produced by the addition of ethylenediamine-di-o hydroxyphenol acetic acid to the growth medium. The density of the biofilm which formed on the base of microtiter plates was dependent on the degree of iron limitation, the stage of the growth cycle, and the nutritional state of the initial inoculum. One repeatedly slime-negative S. epidermidis strain, passaged in tryptic soya broth containing ethylenediamine-di-o-hydroxyphenol acetic acid, expressed high levels of slime after two passages. These observations suggest that iron limitation is one factor that regulates slime production by S. epidermidis. These findings could explain inconsistencies between the in vivo observation that biofilms invariably form on implanted catheters and the in vitro finding that some isolates from catheter-associated infection fail to produce slime. PMID- 8406840 TI - Three-dimensional reconstruction of Coxiella burnetii-infected L929 cells by high voltage electron microscopy. AB - Previous examination of thin sections of L929 cells heavily infected with the Q fever Priscilla isolate by conventional transmission electron microscopy indicated that the rickettsiae resided within multiple vacuoles. The present study using high-voltage electron microscopy and three-dimensional reconstruction revealed that, in heavily infected cells, the rickettsiae, in fact, reside in one multilobed vacuole. As a result of asymmetric cell division, the multilobed vacuole containing the rickettsiae apparently segregates into one daughter cell, while the companion daughter cell emerges parasite free. This likely explains the appearance of naive uninfected cells in long-term-infected (i.e., ca. 2 years) cell populations that had not been supplemented with uninfected L929 host cells. PMID- 8406841 TI - Role of acid tolerance response genes in Salmonella typhimurium virulence. AB - The atp and fur genes are involved in the acid tolerance response of Salmonella typhimurium. An atp::Tn10 mutant was avirulent in the mouse typhoid model when assayed by oral and intraperitoneal routes. However, a fur mutant was completely virulent by the intraperitoneal route. No relevant differences in intracellular survival or invasion rates were observed for the two mutants in macrophages and epithelial cells. These data indicate that separate acid tolerance response genes may have different roles in S. typhimurium virulence. PMID- 8406842 TI - Variant responses of mice to Borrelia burgdorferi depending on the site of intradermal inoculation. AB - C3H/He mice inoculated intradermally at one of two sites with Borrelia burgdorferi responded differently to infection. Shoulder-inoculated mice developed spirochetemia, B. burgdorferi-specific antibody, and arthritis earlier than foot-inoculated mice. Lymphocyte populations derived from spleen tissue were elevated in the shoulder- but not the foot-inoculated mice, and those from lymph nodes were increased in both groups. Lymphocytes derived from blood and spleen tissue showed impaired proliferative responses to all mitogens for shoulder inoculated mice only, whereas proliferation of lymph node cells was not affected, regardless of route. These results demonstrate that the site of initial B. burgdorferi inoculation is an important determinant in the pathogenesis of B. burgdorferi infection. PMID- 8406843 TI - Monocyte antimycobacterial activity before and after Mycobacterium bovis BCG vaccination in Chingleput, India, and London, United Kingdom. AB - Monocytes from purified protein derivative S Mantoux-negative children and young adults inhibited intracellular growth of Mycobacterium microti more in Chingleput than in London. Mycobacterium bovis BCG vaccination did not enhance bacteriostasis with the Indians but did so with the Londoners. No evidence was found for involvement of cytokines such as macrophage-activating factor and granulocyte macrophage colony-stimulating factor in the differences. PMID- 8406844 TI - Identification of the Yersinia enterocolitica urease beta subunit as a target antigen for human synovial T lymphocytes in reactive arthritis. AB - The local T-cell response to bacterial antigens is involved in the pathogenesis of reactive arthritis (ReA). Here, we have identified a 19-kDa antigen of Yersinia enterocolitica O:9 recognized by Yersinia-specific synovial fluid CD4+ T cells in two patients with Yersinia-induced ReA. N-terminal amino acid sequencing of this protein revealed that it was identical to the 19-kDa urease beta subunit of Y. enterocolitica O:9. This protein has previously been shown to be arthritogenic in preimmunized rats after intra-articular injection. Analysis of the T-cell response to this protein showed that it contains several T-cell epitopes, one of which cross-reacts with other enterobacteria not able to induce ReA. This indicates that the arthritogenicity of the 19-kDa antigen is not a property of the 19-kDa protein alone but is dependent on its expression in bacteria able to induce ReA. PMID- 8406845 TI - Mitomycin-induced synthesis of a Shiga-like toxin from enteropathogenic Escherichia coli H.I.8. AB - Escherichia coli H.I.8, an O128 infant diarrhea isolate, produces low titers of a unique Shiga-like toxin (SLT), called SLT-IIva, which is a variant of SLT-II. We investigated induction of toxin synthesis and the putative association of a bacteriophage with toxin synthesis. Induction of broth cultures of strain H.I.8 with mitomycin yielded a 3,000-fold increase in SLT-IIva, production of a colicin, and appearance of a bacteriophage. Southern hybridization demonstrated that the genes for SLT-IIva were not carried by the bacteriophage. PMID- 8406846 TI - An artificial glycoconjugate containing the bisphosphorylated glucosamine disaccharide backbone of lipid A binds lipid A monoclonal antibodies. AB - Monoclonal antibodies (MAbs) against lipid A, the endotoxic component of lipopolysaccharide (LPS) of gram-negative bacteria, are presently discussed as therapeutic agents against lethal gram-negative infections; however, their binding specificities are controversial. We have isolated from the LPS of Escherichia coli J-5 the 1,4'-bisphosphorylated beta 1-->6-linked glucosamine disaccharide backbone of its lipid A moiety, which was covalently linked to bovine serum albumin. It was shown by solid-phase enzyme immunoassay that one antibody (MAb A6) bound equally well to the glycoconjugate and synthetic E. coli type lipid A over a broad range of antigen concentrations whereas two other MAbs (IC3 and S1-15) bound better to the conjugate at low antigen concentrations and better to the lipid A when high concentrations of antigen were used. This proves in a direct way that there exist lipid A MAbs with different specificities which bind to epitopes in the hydrophilic backbone of lipid A and which do not require the presence of fatty acids. PMID- 8406847 TI - The 39-kilodalton protein of Borrelia burgdorferi: a target for bactericidal human monoclonal antibodies. AB - Three human monoclonal immunoglobulin M antibodies against Borrelia burgdorferi, obtained from in vitro-stimulated peripheral blood lymphocytes, reacted in Western blots (immunoblots) with a prominent 39-kDa peptide and a faint band of approximately 66 kDa. Two of these antibodies showed bactericidal activity without addition of complement. All three antibodies were reactive in an enzyme immunoassay with cloned P39 (W.J. Simpson, M.E. Schrumpf, and T.G. Schwan, J. Clin. Microbiol. 28:1329-1337, 1990), suggesting that the target molecule of these antibodies is identical to the P39 protein. In addition, the majority of supernatants from human lymphocytes stimulated in vitro with crude B. burgdorferi antigen reacted in this assay, demonstrating that P39, although a minor component of B. burgdorferi, is an immunodominant antigen in these spirochetes. A fourth monoclonal antibody, reacting with OspA, also exhibited bactericidal activity. PMID- 8406848 TI - R plasmid in Escherichia coli O103 coding for colonization of the rabbit intestinal tract. PMID- 8406849 TI - Phlogistic properties of peptidoglycan-polysaccharide polymers from cell walls of pathogenic and normal-flora bacteria which colonize humans. AB - PG-PS polymers which can induce experimental chronic inflammation in joints and other tissues can be isolated from the cell walls of human pathogens, such as group A streptococci, as well as from certain indigenous bacterial species which colonize the human intestinal tract. The structural and biological properties that are required for cell wall fragments to express this remarkable activity are still not well defined, but polymer size, resistance to tissue enzymes, and capacity to sustain activation of complement, macrophages, neutrophils, and T cells are properties associated with the most active preparations. There is increasing evidence that PG-PS structures with arthropathogenic activity occur in the human intestinal lumen and that these polymers can be translocated systemically. These observations support the concept that PG-PS, derived from a variety of bacterial species, can be part of the etiology of rheumatoid arthritis and other chronic inflammatory diseases. Since the PG component provides a common element to which all individuals are exposed, it follows that susceptibility is related to efficiency of disposal of bacterial cell wall debris, as well as to cytokine networks and immune cell function (51). PMID- 8406850 TI - Organization of two invariant surface glycoproteins in the surface coat of Trypanosoma brucei. AB - The surface coat of Trypanosoma brucei, formed by about 10(7) molecules of the membrane-form variant surface glycoprotein (mfVSG) per cell, is generally considered to constitute a barrier against the access of antibodies directed to invariant surface proteins. The recent characterization of two invariant surface glycoproteins (ISGs) with apparent molecular masses of 65 and 75 kDa (ISG65 and ISG75; 70,000 and 50,000 molecules per cell, respectively), which are both predicted to be composed of large extracellular domains, single transmembrane alpha-helices, and small intracellular domains, enabled a critical test of this hypothesis. Although ISG65 is distributed over the entire surface of the parasites, it is not accessible to antibodies or to the proteinase trypsin in live cells provided the mfVSG is also proteinase resistant. ISG75 is similarly distributed; its accessibility to antibodies depends on the expressed mfVSG, and it is sensitive to trypsin in a variant clone in which the mfVSG is proteinase resistant. Vaccination experiments using recombinant proteins to a mixture of the native ISGs were unsuccessful. ISG65 but not ISG75 elicited an antibody response in chronically infected mice. The results strengthen the view of the protective properties of the variant surface glycoprotein coat by steric hindrance and suggest that additional factors such as low abundance or low immunogenicity of invariant surface proteins may prevent a control of the disease by the humoral immune response. PMID- 8406851 TI - High levels of interleukin-8 in the blood and alveolar spaces of patients with pneumonia and adult respiratory distress syndrome. AB - There is ample experimental evidence that polymorphonuclear neutrophils (PMN) play a critical role in the pathogenesis of the adult respiratory distress syndrome (ARDS). Since interleukin-8 (IL-8) is a strong chemotactic factor for PMN, we measured IL-8 levels in plasma and bronchoalveolar lavage (BAL) fluid of 18 patients, 12 with ARDS and 6 with severe pneumonia uncomplicated by ARDS, all of whom had an increased number of PMN in BAL fluid. Seven healthy subjects served as controls. We found elevated levels of IL-8 in the alveolar spaces of all patients tested. Elevated BAL IL-8 levels were related to a fatal outcome and the presence of shock and correlated with a general clinical severity index (simplified acute physiological score). BAL fluid levels of IL-8 were significantly higher in patients with ARDS than in patients with pneumonia. In plasma, IL-8 levels were increased similarly in all patients and did not correlate with survival or the presence of shock. The BAL fluid-to-plasma ratio of IL-8 was significantly greater than that of tumor necrosis factor alpha, indicating higher local production of IL-8. Moreover, the presence of a primed subpopulation of blood PMN with respect to H2O2 production indicates that IL-8 may contribute to the neutrophil-mediated process in the pathogenesis of ARDS and pneumonia. PMID- 8406852 TI - Mechanisms of adherence of Candida albicans to cultured human epidermal keratinocytes. AB - We established an in vitro adherence model with primarily cultured human keratinocytes as target cells which allows for the investigation of the molecular mechanisms that are responsible for Candida albicans host cell attachment in the initiation of cutaneous candidosis. The extent of C. albicans binding to cultured human keratinocytes was dependent on the yeast inoculum size and the incubation temperature. Heat and paraform-aldehyde treatment of yeasts completely abolished the binding activity of C. albicans. Of the different Candida species tested, C. albicans was by far the most adhesive species. C. albicans adherence was blocked by the acid protease inhibitor pepstatin A and the metabolic inhibitor sodium azide. The latter, however, was much less effective when yeasts were preincubated, suggesting that sodium azide was mainly acting on the keratinocytes. The extracellular matrix protein fibronectin was slightly inhibitory, whereas the fibronectin-derived peptides RGD and RGDS were not able to prevent attachment. PepTite-2000, another RGD-containing synthetic peptide, reduced C. albicans adherence by a margin of 25% (P < 0.005). CDPGYIGSR-NH2, which is a synthetic adhesive peptide derived from the laminin B chain, was much more efficient in its inhibitory activity than the RGD peptides and reduced C. albicans adherence to cultured human keratinocytes up to 76% (P < 0.001). Laminin itself and the synthetic pentapeptide YIGSR were less active. A dose-dependent reduction in adherence was also observed with collagen type III. Additionally, saccharides were tested for their potential to inhibit C. albicans attachment to keratinocytes. The most potent competitive saccharide inhibitors of C. albicans adherence to human keratinocytes were the amino sugars D-(+)-glucosamine and D (+)-galactosamine with one isolate of C. albicans (4918) and D-(+)-glucosamine and alpha-D-(+)-fucose with another C. albicans isolate (Sp-1). Collectively, our data suggest the existence of multiple molecular mechanisms such as protein protein, lectin-carbohydrate, and yeast-yeast coaggregational interactions that are responsible for optimal C. albicans attachment to cultured human keratinocytes. PMID- 8406853 TI - Epithelial cells secrete the chemokine interleukin-8 in response to bacterial entry. AB - Bacterial invasion of mucosal surfaces results in a rapid influx of polymorphonuclear leukocytes. The chemotactic stimulus responsible for this response is not known. Since epithelial cells are among the first cells entered by many enteric pathogens, we investigated the ability of epithelial cells to provide an early signal for the mucosal inflammatory response through the release of chemotactic cytokines. As shown herein, the chemokine interleukin-8 (IL-8), a potent chemoattractant and activator of polymorphonuclear leukocytes, was secreted by intestinal and cervical epithelial cells in response to bacterial entry. Moreover, a variety of different bacteria, including those that remain inside phagosomal vacuoles, e.g., Salmonella spp., and those that enter the cytoplasm, e.g., Listeria monocytogenes, stimulated this response. Increased IL-8 mRNA levels could be detected within 90 min after infection. Neither bacterial lipopolysaccharide nor noninvasive bacteria, including Escherichia coli and Enterococcus faecium, induced an IL-8 response. Moreover, tumor necrosis factor alpha, which is known to be expressed by some epithelial cells, was not detected in the culture supernatants after bacterial entry, and addition of anti-tumor necrosis factor alpha antibodies had no effect on the IL-8 response following bacterial entry. These data suggest the novel concept that epithelial cells serve as an early signaling system to host immune and inflammatory cells in the underlying mucosa following bacterial entry. PMID- 8406854 TI - Antigenic variation of immunoglobulin A1 proteases among sequential isolates of Haemophilus influenzae from healthy children and patients with chronic obstructive pulmonary disease. AB - Considerable antigenic heterogeneity has been identified among Haemophilus influenzae immunoglobulin A1 (IgA1) proteases, and this study increases the number of antigenic types to more than 30. To address the role played in vivo by this polymorphism, sequential H. influenzae isolates from three healthy children and three patients with chronic obstructive pulmonary disease (COPD) were examined. Healthy children showed a frequent clonal exchange, with each replacing clone expressing an antigenic type of IgA1 protease not previously encountered. In contrast, COPD patients were colonized by a single clone for a significantly longer period. In one COPD clone, a change occurred in IgA1 protease cleavage specificity and antigenic properties. In conclusion, frequent exchange of clones expressing antigenically different IgA1 proteases seems to be the principal mechanism by which H. influenzae evades the immune response of healthy children against IgA1 protease. The results support the view that IgA1 protease activity is important for successful colonization of H. influenzae on mucosal membranes. PMID- 8406855 TI - Novel antigens expressed by Aeromonas salmonicida grown in vivo. AB - Virulent and avirulent Aeromonas salmonicida strains grown inside intraperitoneal implants in Rainbow trout (Oncorhynchus mykiss) were examined for unique antigen expression. Western blots (immunoblots), performed with immune rabbit serum raised against in vivo-grown cells, revealed several unique antigens. With the exception of lipopolysaccharide (LPS), these novel antigens were destroyed after proteinase K treatment. The majority of these antigens were not induced in vitro in response to either iron limitation or anaerobiosis. In addition, electron microscopy demonstrated the presence of a putative capsule on in vivo-grown cells. Purification and fractionation of this carbohydrate material from cells grown in carbon-rich synthetic media resulted in the isolation and separation of an antigenically distinct LPS not seen with cells grown in standard media. Antiserum raised against in vivo-grown cells recognized both this LPS and the typical LPS of A. salmonicida apparent in in vitro-grown cells. Antiserum raised against in vitro-grown cells recognized only the LPS expressed in vitro. Antiserum directed against in vivo-grown cells was approximately 10 times more sensitive than serum directed against in vitro-grown cells in detecting A. salmonicida in infected fish kidney tissue. PMID- 8406856 TI - Identification and cloning of a fur homolog from Neisseria gonorrhoeae. AB - The promoter region of the major iron-regulated protein of Neisseria gonorrhoeae, Fbp, has two regions that exhibit homology with the Escherichia coli consensus Fur-binding sequences. Gel retardation assays suggested that purified E. coli Fur bound to two sites within the Fbp promoter. The presence of a gonococcal Fur homolog was suggested by Southern hybridization under conditions of low stringency, which revealed a DNA locus that exhibited homology to the E. coli fur gene. Oligonucleotides derived from the conserved regions of fur genes of extremely diverse bacteria were used to amplify a 140-bp fragment of a putative gonococcal fur gene. This fragment was used to identify clones containing the entire gonococcal fur gene. After sequencing the gonococcal fur gene and its promoter region, we found that gonococcal Fur exhibited 50% identity with E. coli Fur at the amino acid level; however, it complemented two E. coli Fur- mutants. The presence of a Fur homolog in N. gonorrhoeae suggests that Fur-regulated genes are widely distributed among extremely diverse bacteria. PMID- 8406857 TI - Local immunity in lung-associated lymph nodes in a murine model of pulmonary histoplasmosis. AB - Local immunity against acute pulmonary histoplasmosis was studied in the lung associated lymph nodes of normal nonimmune mice infected intratracheally with live Histoplasma capsulatum yeasts. The phenotypes and distribution of cells in lung-associated lymph nodes and spleens were determined by flow cytometry. In addition, the immune responsiveness of these cells was evaluated by in vitro blastogenesis. Anti-H. capsulatum antibodies in serum and H. capsulatum antigen in tissue were measured by immunoassays. Cellular immune responses were greater in the lymph nodes than in the spleens. In lymph nodes 7 days after infection, a marked increase in the number of B lymphocytes caused the percentage to rise to 43%, compared with 26% in controls, and it remained elevated throughout the course of infection. A CD3+ cell that did not express CD4 or CD8 increased in number until it constituted 21% of lymph node cells, compared with 5% in controls, by day 14. The numbers of CD4+ and CD8+ T lymphocytes were modestly increased from days 7 to 35, but their percentages dropped because of the greater numbers of B lymphocytes and CD3+4-8- cells. Macrophages consistently constituted 2 to 3% of lymph node cells during the study. In spleens 7 days after infection, the percentage of macrophages in infected mice rose to 21%, compared with 9% in controls, but the total spleen cell number did not increase until day 14, when all cell subsets were nearly double in number. The in vitro blastogenic response of lymph node cells to H. capsulatum peaked at day 7, but spleen cell response was minimal during the course of infection. Histoplasma-specific serum immunoglobulin G antibodies reached peak levels by day 21 and remained high to the end of the study. In contrast, levels of antigen-specific immunoglobulin M antibodies were very low. These data suggest that antigen-specific immune responses occur in lung-associated lymph nodes and that this draining lymph node response may be an important component in host defense against Histoplasma lung infection. PMID- 8406858 TI - Genetic control of resistance to Mycoplasma pulmonis infection in mice. AB - The differences in susceptibility of various inbred strains of mice to a highly pathogenic strain of Mycoplasma pulmonis CT (T2) has been known for some time. We assessed the genetic control of resistance to T2 infection. Tracheolung lavage samples and lungs of mice were assessed for T2 organisms after intratracheal injection of T2. We found that H-2b (C57BL/6 (B6) and H-2k B10.BR mice were resistant, whereas H-2b A.By, H-2k C3H/Bi, H-2k C3H/HeJ (C3H), and H-2b BALB.B mice were susceptible. We also typed individual B6C3F2 mice for H-2 and for resistance to T2 and observed that resistance to T2 infections is controlled by a single dominant gene not linked to H-2. Histologic examination revealed severe lung lesions typical of M. pulmonis infections in susceptible C3H mice, in contrast to minimal lung lesions in resistant B6 mice. No significant titers of local or systemic antimycoplasma antibodies were detected in either resistant or susceptible mice at 5 days postinfection. Macrophages taken from uninfected B6 or C3H mice failed to inhibit growth of T2 in vitro. However, macrophages from B6 mice did inhibit growth of T2 much better than C3H macrophages when harvested on day 5 of infection. Thus, there is an association between activation of macrophage bactericidal function and genetic resistance to growth of T2 organisms. PMID- 8406859 TI - Expression of monocyte chemoattractant protein 1 in human inflamed gingival tissues. AB - Gingival inflammation is initiated by bacterial colonization on the tooth surface. It is characterized by infiltration of mononuclear cells, a common feature of many forms of chronic inflammation. Monocyte chemoattractant protein 1 (MCP-1) is the predominant monocyte chemoattractant secreted by a variety of different cells in vitro. For this report, we examined MCP-1 expression in bacterially induced gingival inflammation by immunohistochemistry and in situ hybridization. The cell types expressing MCP-1 are identified as vascular endothelial cells and monocytes/macrophages. Correlation analysis shows that the number of cells expressing MCP-1 is related to the degree of inflammation. Our finding that MCP-1 is expressed in inflamed gingival tissue suggests that MCP-1 plays an important role in the recruitment of monocytes and amplification of inflammatory signals in bacterially induced inflammation. PMID- 8406860 TI - Development of mucosal protection against the heat-stable enterotoxin (ST) of Escherichia coli by oral immunization with a genetic fusion delivered by a bacterial vector. AB - An LT-B-ST (LT-B/ST) fusion peptide was constructed by genetically joining the 5' terminus of a synthetic gene coding for the heat-stable enterotoxin (ST) of Escherichia coli to the 3' terminus of the gene coding for the binding subunit of the heat-labile enterotoxin (LT-B) of E. coli. An eight-amino-acid, proline containing linker was included between the LT-B and ST moieties. An aroA mutant of Salmonella dublin transformed with a plasmid carrying this genetic construct was shown to express a fusion peptide with antigenic determinants of both LT-B and ST. Mice were immunized orally with this strain or with a control strain expressing just LT-B from the same plasmid. Sera and mucosal secretions were obtained and analyzed for the presence of serum immunoglobulin G and mucosal immunoglobulin A that were able to recognize LT-B and ST by enzyme-linked immunosorbent assay (ELISA) and, more importantly, were able to neutralize native ST in the suckling mouse assay. Sera and mucosal secretions from animals immunized with the strain expressing the LT-B/ST fusion exhibited detectable ELISA reactivity against LT-B but not against native ST. However, even in the absence of detectable ELISA reactivity, both sera and mucosal secretions from these animals were able to neutralize the biological activity of native ST in the suckling mouse assay. These findings are important because they demonstrate the development of mucosal protection against ST by oral immunization with a genetic fusion delivered by a bacterial vector. PMID- 8406861 TI - Enhancing effect of cholera toxin on interleukin-6 secretion by IEC-6 intestinal epithelial cells: mode of action and augmenting effect of inflammatory cytokines. AB - Oral administration of cholera toxin (CT) induces a strong mucosal immune response to CT as well as having a potent adjuvant effect. Since one of the first cell types to encounter CT during cholera infection or after oral administration is the epithelial cell, we studied the effect of CT on interleukin-6 (IL-6) secretion by the rat intestinal epithelial cell line IEC-6. CT was found to rapidly enhance IL-6 secretion and IL-6 gene expression by these cells. The addition of dibutyryl cyclic AMP (cAMP) to cultures of IEC-6 cells had little effect on IL-6 secretion, yet mRNA levels were elevated, suggesting that the response may have been regulated by cAMP. Purified B subunit of CT did not significantly enhance IL-6 secretion or mRNA expression. CT and transforming growth factor beta 1 synergistically enhanced IL-6 secretion in IEC-6 cells. The addition of CT with either IL-1 beta or tumor necrosis factor alpha gave even greater synergistic enhancement of IL-6 secretion, and dibutyryl cAMP could mimic CT's synergy with IL-1 beta. These results indicate that the intestinal epithelial cell is capable of secreting high levels of IL-6 after encountering CT, especially in the presence of inflammatory cytokines. This high level of IL-6 secretion could be a very important component of the mucosal immune response to CT and may also account for a portion of the adjuvant effect of CT. PMID- 8406862 TI - Bacterial cell wall polymers (peptidoglycan-polysaccharide) cause reactivation of arthritis. AB - Intraperitoneal (i.p.) injection of peptidoglycan-polysaccharide derived from group A streptococci (PG-APS) causes chronic arthritis with spontaneous remissions and exacerbations. We hypothesized that, following i.p. injection, PG APS released from hepatic stores mediated spontaneous recurrences of arthritis. We tested whether transplanted livers with large amounts of PG-APS were able to reactivate quiescent arthritis. Saline-loaded (group 1) or PG-APS-loaded (group 2) livers were transplanted into rats which had been injected intra-articularly 10 days earlier with PG-APS in one joint and saline in the other. A comparison was made with the arthritis that occurred in rats injected i.p. with PG-APS which did not receive transplants (group 3). Arthritis was monitored by serial measurement of joint diameters. Transplantation of saline-loaded livers (group 1) caused no reactivation of arthritis. However, transplantation of PG-APS-loaded livers (group 2) reactivated arthritis (P < 0.0001). Injection of PG-APS i.p. (group 3) induced the most-severe arthritis. PG-APS levels in plasma decreased with time, and PG-APS accumulated in the spleen in groups 2 and 3. Plasma and hepatic levels of PG-APS in rats injected i.p. with PG-APS were greater than levels in rats transplanted with PG-APS-loaded livers, which in turn were greater than levels in rats with saline-loaded livers. Plasma tumor necrosis factor did not correlate with recurrence of arthritis. Transplantation with PG-APS-loaded livers induced reactivation of arthritis in preinjured joints. The extent of arthritis was proportional to hepatic PG-APS content. Reactivation of arthritis may be mediated by slow release of liver-sequestered PG-APS or cytokines (not tumor necrosis factor) released by the liver. PMID- 8406863 TI - The eae gene of Citrobacter freundii biotype 4280 is necessary for colonization in transmissible murine colonic hyperplasia. AB - Transmissible murine colonic hyperplasia is characterized by proliferation of anchored stem cells in the mucosa of the descending colon of laboratory mice and is caused by Citrobacter freundii biotype 4280. This bacterium produces attaching and effacing lesions in the descending colon prior to the onset of gross hyperplasia. By mutational analysis, the chromosomal eae gene of C. freundii biotype 4280 was shown to be necessary for colonic colonization. Conversely, bacteria cured of a 65-kb plasmid, which was identified in C. freundii biotype 4280, were not attenuated for colonic colonization or for the induction of colonic hyperplasia. PMID- 8406864 TI - Increased sensitivity of gonococcal pilA mutants to bactericidal activity of normal human serum. AB - PilA is a pleiotropic transcriptional regulator in Neisseria gonorrhoeae, encoded by an essential gene, pilA. It regulates pilin gene expression and stress response and it is implicated in gonococcal adaptation to external signals. All these phenomena may participate in gonococcal virulence. In this report, I tested the role of PilA in another aspect of gonococcal virulence, resistance to the bactericidal effect of normal human serum. Gonococcal mutants with impaired PilA function were more susceptible to the bactericidal effect of normal human serum than the isogenic wild-type strain. However, the major outer membrane protein and the lipooligosaccharide, targets for complement-mediated killing by the serum, were unchanged in the mutants. I discuss the role of PilA in modulating gonococcal sensitivity and resistance to normal human serum. PMID- 8406865 TI - Characterization of neuraminidases produced by various serotypes of Pasteurella haemolytica. AB - Neuraminidases produced by 16 strains of Pasteurella haemolytica (serotypes 1 to 16) were characterized by molecular weight, antigenic identity, and substrate specificity. After growth in a chemically defined medium, stage I (lyophilized) culture supernatants were assayed for activity with N-acetylneuramin lactose, human alpha-1-acid glycoprotein, fetuin, colominic acid, and bovine submaxillary mucin. Neuraminidase produced by P. haemolytica serotype A1 (Ph A1) was purified by a combination of salt fractionation, ion-exchange chromatography on DEAE Sephacel, and gel filtration on Sephadex G-200. Purified Ph A1 neuraminidase was used to immunize rabbits, and the resultant antiserum reduced the activity of Ph A1 neuraminidase by 46%. This antiserum also reduced the activity of neuraminidase produced by the other serotypes by between 15 and 66%. Molecular weight estimates of the neuraminidases produced by the various serotypes were obtained by gel filtration chromatography on Sephadex G-200. Fifteen of the 16 serotypes examined produced a neuraminidase with a molecular weight of approximately 150,000 to 200,000. One serotype (serotype 11) produced no material with neuraminidase activity. In addition, all 15 high-molecular-weight neuraminidases showed similar substrate specificities. That is, they were all most active against N-acetylneuramin lactose and least active against bovine submaxillary mucin. On the basis of these results, it appears that the high molecular-weight neuraminidases produced by the different P. haemolytica serotypes are quite similar. PMID- 8406866 TI - Three contiguous lipoprotein genes in Pasteurella haemolytica A1 which are homologous to a lipoprotein gene in Haemophilus influenzae type b. AB - An Escherichia coli clone carrying the recombinant plasmid pPH24 has been found to express highly immunoreactive antigens of Pasteurella haemolytica A1. Two or three antigens of approximately 30 kDa were located to both the inner and outer membranes of the E. coli clone and in P. haemolytica A1. From the insert DNA of 8.2 kbp on pPH24, a fragment of 4.6 kbp was found to code for these antigens. Nucleotide sequence analysis of the 4.6-kbp DNA identified three genes (designated plpA, -B, and -C) arranged in tandem which code for three proteins, Plp1, -2, and -3, with predicted molecular masses of 30.1, 30.3, and 29.0 kDa, respectively. Comparison of the nucleic acid sequence of plpA, -B, and -C with GenBank sequences showed extensive homology with a Haemophilus influenzae 28-kDa lipoprotein gene. [14C]palmitate labelling coupled with glybomycin inhibition experiments showed that Plp1, -2, and -3 are also lipoproteins. In addition, plpA, -B, and -C were found to be present only in A biotypes of P. haemolytica by Southern blot analysis. Since Plp1, -2, and -3 were found to be antigenic components in a culture supernatant vaccine, they could be candidates for further investigation as vaccine components. PMID- 8406867 TI - Inhibition of glucosyltransferase activities of Streptococcus mutans by a monoclonal antibody to a subsequence peptide. AB - Preliminary analysis indicated that a 19-amino-acid peptide sequence (435 to 453 of GtfC) within a highly conserved region of the glucosyltransferases of the cariogenic streptococci might be functionally important (J.-S. Chia, S.-W. Lin, T.-Y. Hsu, J.-Y. Chen, H.-W. Kwan, and C.-S. Yang, Infect. Immun. 61:1563-1566, 1993). To obtain antipeptide monoclonal antibodies (MAbs), the 19-amino-acid peptide was conjugated to bovine serum albumin and used as an antigen in BALB/c mice. Six immunoglobulin G-secreting hybridoma clones, CJSm18-S1 to -S6, specifically reacted with this peptide and with purified GtfC and GtfD but not with bovine serum albumin in an enzyme-linked immunosorbent assay. The concentrated hybridoma supernatant of all six MAbs inhibited GtfC enzymatic activity but failed to inhibit GtfD, although GtfD contains the same peptide sequence. Further analysis of a purified immunoglobulin G2b MAb from one of the clones, CJSm18-S3, confirmed that this MAb specifically inhibited GtfC enzymatic activity for insoluble-glucan synthesis in a dose-dependent manner. CJSm18-S3, even at high concentrations, had no effect on GtfD, which synthesizes water soluble glucan exclusively. Furthermore, the in vitro sucrose-dependent adherence of Streptococcus mutans was also inhibited by CJSm18-S3 in a dose-dependent manner. Our results indicate that the peptide containing the N-terminal conserved region of glucosyltransferases is functionally important for both enzymatic activity and bacterial adherence. PMID- 8406868 TI - Passive immunizing activity of sera from mice infected with Borrelia burgdorferi. AB - A single injection of serum from C3H mice at 90 days after intradermal inoculation with 10(4) Borrelia burgdorferi spirochetes protected naive mice when administered subcutaneously at -18 h relative to intradermal challenge inoculation with 10(4) B. burgdorferi spirochetes. When immune serum was given at intervals (-18, 0, +24, +48, and +96 h) relative to intradermal challenge with 10(4) B. burgdorferi spirochetes, it was protective if given before or at the time of challenge but not at later times. Protection with 90-day serum given at 18 h was effective at dilutions up to 1:32, but not 1:64, on the basis of culture or disease at either 5 or 15 days after challenge. Passive immunizing activity was also present in sera from mice at 21 days after intradermal challenge with 10(4), 10(2), or 10(1) spirochetes, indicating that the immunizing component was not dose dependent and probably not related to antibody to outer surface protein A. Passive immunizing titers of sera from mice at days 1, 15, 30, 90, 180, and 360 after intradermal B. burgdorferi inoculation appeared as early as day 15, were highest on day 30, and then declined progressively on days 90, 180, and 360. Immunizing titers of sera from mice at 360 days after intradermal B. burgdorferi inoculation were identical in passively immunized mice challenged with the original inoculum or with B. burgdorferi isolated at 360 days after inoculation, suggesting that there was no antigenic discrimination between the original inoculum and late isolates. These results suggest that protective antibody is produced early in the course of B. burgdorferi infection and is unrelated to antibody to outer surface protein A. In addition, the decline of protective serum titers over time despite persistent infection suggests that the antigens eliciting the protective response are either modified or suppressed, but antigenic modification could not be demonstrated. PMID- 8406869 TI - Proliferative and cytokine responses to a major surface glycoprotein of Pneumocystis carinii. AB - Naturally derived T-cell responses by rats to a 120-kDa major surface glycoprotein (MSG) of rat-derived Pneumocystis carinii were analyzed in vitro. Specific cytokines elicited by the T-cell response to the MSG were also identified. MSG was purified from rat-derived P. carinii by three different techniques: lectin affinity chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by electroelution, and size-exclusion high performance liquid chromatography. The cell-mediated immunity of spleen cells isolated from Lewis rats with and without natural exposure to P. carinii to the purified MSG was studied. Exposure to P. carinii was monitored by the presence or absence of serum antibodies to P. carinii antigens by Western blotting (immunoblotting). A T-cell proliferative response to the MSG was identified only with spleen cells isolated from rats exposed to P. carinii and peaked at 4 days. Flow cytometric analysis revealed that the percentage of CD4 cells was significantly increased during the proliferative response to MSG. MSG also elicited secretion of tumor necrosis factor alpha, interleukin-1, and interleukin 2, with peak activity of these cytokines occurring after 12, 24, and 48 h, respectively, of culture. These findings suggest that MSG is important in host T cell recognition of and immune response to P. carinii by recruitment of inflammatory cells and cytokine production. PMID- 8406870 TI - Cloning, molecular characterization, and functional activity of Schistosoma japonicum glyceraldehyde-3-phosphate dehydrogenase, a putative vaccine candidate against schistosomiasis japonica. AB - We report the cloning, molecular characterization, and purification of functionally active recombinant glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the human bloodfluke Schistosoma japonicum. The GAPDH homolog from the related species Schistosoma mansoni has shown correlation of antibody titer to resistance to reinfection. A 1,164-bp cDNA (C1) was isolated from an S. japonicum lambda ZapII cDNA expression library immunoscreened with hyperimmune rabbit serum raised against soluble adult S. japonicum proteins. The open reading frame of C1 encodes a protein of 338 amino acids exhibiting 90% identity to the amino acid sequence of S. mansoni GAPDH. The inferred molecular mass of the protein is 36,589 daltons, and in vitro translation of the cDNA with [35S]methionine produced a radiolabelled band of the predicted size. Antibodies to C1 selected from hyperimmune rabbit serum by affinity purification recognized an S. japonicum protein doublet of 37 kDa but did not cross-react with a corresponding protein in S. mansoni extracts. The S. japonicum GADPH appears to be translated from a single mRNA encoded by a single-copy gene. After subcloning in the QIAexpress vector pQE-10 and subsequent expression, the recombinant protein was purified under nondenaturing conditions and shown to exhibit functional GAPDH enzymatic activity. PMID- 8406871 TI - Molecular characterization of the 98-kilodalton iron-regulated outer membrane protein of Neisseria meningitidis. AB - When grown under iron limitation, Neisseria meningitidis expresses several additional outer membrane proteins (OMPs), which were studied to assess their vaccine potential. Two monoclonal antibodies were obtained against a 98-kDa OMP of strain 2996 (B:2b:P1.2). Cross-reactivity studies revealed that the two antibodies reacted with 44 and 42 of 74 meningococcal strains, respectively. The antibodies did not block the binding of transferrin or lactoferrin to intact cells. The structural gene for the protein, tentatively designated iroA, was isolated and sequenced. Computer analysis revealed homology to the ferric siderophore receptors in the outer membrane of Escherichia coli and to gonococcal transferrin-binding protein 1 (TbpA). The high degree of cross-reactivity and the results of Southern blot analyses, which showed that the iroA gene is also present in strains that did not react with the monoclonal antibodies, suggest that the 98-kDa OMP is well conserved among meningococci and that it is a suitable vaccine candidate. However, the antibodies were not bactericidal in an in vitro assay with human complement. PMID- 8406872 TI - Humoral immune response to the class 3 outer membrane protein during the course of meningococcal disease. AB - We have determined the amounts of specific anti-class 3 outer membrane protein antibodies of immunoglobulin G (IgG), IgM, and IgA isotypes in patient sera during the course of meningococcal disease by using purified class 3 protein as the sensitizing antigen in an enzyme-linked immunosorbent assay. The class 3 protein was obtained from a variant of strain 44/76 (B:15:P1.7,16) lacking class 1 and class 4 outer membrane proteins. Serum samples from 25 patients with systemic meningococcal disease caused by organisms of various serotypes were collected during the course of disease. Seven of these patients had been immunized with a meningococcal outer membrane vesicle vaccine made from strain 44/76 prior to disease. An increase in specific anti-class 3 (type 15) outer membrane protein IgG antibodies was demonstrated in 22 of 25 patients (88%), regardless of the serotype of the infecting strain. This indicates that the specific anti-class 3 antibodies were reacting in part with epitopes not determined by the monoclonal antibodies used for serotyping. A considerable heterogeneity in antibody levels and IgG subclass response was seen. Most patients had low levels of anti-class 3 antibodies during the acute illness, with antibodies peaking during the second week of disease and returning to near baseline in sera collected 6 to 12 months after the onset of the disease. The majority of the specific anti-class 3 IgG antibodies were shown to bind to surface-exposed epitopes on the whole bacteria and to belong to IgG1 and IgG3. The highest anti-class 3 IgG peak levels were seen in patients infected with strains of the homologous serotype after vaccination with the meningococcal outer membrane vesicle vaccine, suggesting an anamnestic response. However, these patients were not protected from meningococcal disease after immunization. PMID- 8406873 TI - Pathology of congenital syphilis in rabbits. AB - We have developed a model for congenital syphilis in the rabbit. This report provides additional information on newborn tissue pathology in animals that were infected in utero. A total of 35 pregnancies were evaluated, each containing 6 to 12 newborns. In the infected group, the mortality was approximately 50%; of the live newborns, half appeared normal and half were hyperreflexic, weak, and runty. Gross pathology in the sickly newborns was quite prevalent and involved enlarged spleens with isolated spots of necrosis; enlarged livers that were overtly congested and hemorrhagic and had numerous granular, white spots; and brains with hemorrhage in the occipital area. Histopathology was apparent in different tissues. Lymphocytes, plasma cells, and vacuolated macrophages were prominent in livers, spleens, brains, and bones. A few actively motile treponemes were visualized by dark-field microscopy within extracts of spleen and within cerebrospinal fluid. Low numbers of treponemes were also demonstrated in sections of brain and liver by using the Warthin-Starry silver stain technique. Blood hematocrits were decreased, and extramedullary hematopoiesis was prominent within spleens and livers; this is consistent with anemia. This rabbit model exhibits many of the same pathologic features commonly found in human congenital syphilis. PMID- 8406874 TI - Cytokine profiles associated with induction of the anticryptococcal cell-mediated immune response. AB - Previous studies with a murine model have shown that immunization with cryptococcal culture filtrate antigen (CneF) emulsified in complete Freund adjuvant (CFA) induces two populations of anticryptococcal reactive CD4+ T cells. One population (TDH cells) transfers anticryptococcal delayed-type hypersensitivity (DTH), and the other population (Tamp cells) amplifies the anticryptococcal DTH response of given to recipient mice at the time of immunization of the recipient. Treatment of mice with cyclosporin A (CsA) ablates the induction of Tamp cells but not TDH cells. The present study focused on assessing the cytokines produced by spleen cells taken from CsA-treated and control (solvent-treated) mice at days 1, 2, 4, and 6 after immunization. Supernatants from the spleen cells cultured in vitro for 24 or 48 h in medium alone or with CneF, concanavalin A, or phorbol 12-myristate 13-acetate plus calcium ionophore were assessed for the presence of interleukin-2 (IL-2), gamma interferon (IFN-gamma), IL-4, IL-5, and tumor necrosis factor. Spleen cells from CneF-CFA-treated mice produced IL-2 and IFN-gamma, but not IL-4 or IL-5, constitutively and in response to CneF, indicating that CneF-CFA induces a Th1 response. Tumor necrosis factor was not produced. Anticryptococcal TDH cells developed in spleens in which there were low levels of IFN-gamma and IL-2 (CsA treated, immunized mice), whereas anticryptococcal Tamp cells along with TDH cells matured in spleens in which production of IFN-gamma and IL-2 was high (solvent-treated, immunized mice). The data also suggest that IL-2 and IFN-gamma produced by Tamp cells early after adoptive transfer are influential in the development of the amplified anticryptococcal DTH response that has been observed in Tamp cell-recipient mice. PMID- 8406875 TI - Stimulation of protective antibodies against type Ia and Ib group B streptococci by a type Ia polysaccharide-tetanus toxoid conjugate vaccine. AB - Antisera elicited by type Ia group B streptococci (GBS) contain antibodies that react with both type Ia and type Ib strains. Previous studies suggested that antibodies elicited by type Ia organisms recognized a carbohydrate antigen or epitope common to Ia and Ib strains. We now report the synthesis and immunogenicity testing of a type Ia polysaccharide-tetanus toxoid (Ia-TT) conjugate vaccine. Ia-TT elicited type Ia polysaccharide-specific immunoglobulin G antibodies in all three of the rabbits inoculated. In competitive enzyme-linked immunosorbent assay, these antibodies reacted with high affinity to type Ia polysaccharide and with lower affinity to the structurally related GBS type Ib polysaccharide. Despite the lower binding affinity of the Ia-TT-induced antibodies for the type Ib polysaccharide, Ia-TT antiserum opsonized not only type Ia GBS but also type Ib GBS for killing by human blood leukocytes. Ia-TT antiserum was also evaluated in a mouse model designed to test the efficacy of maternal antibodies in protecting neonates against GBS infection. Pups born to dams that had received Ia-TT antiserum were protected against lethal challenge with either type Ia or Ib GBS. These studies using a polysaccharide-protein conjugate as an immunogen support the view that the carbohydrate immunodeterminant recognized on Ib strains by Ia antisera is a common epitope contained within the structurally related Ia and Ib capsular polysaccharides. Although antibodies elicited by Ia-TT had protective activity against both Ia and Ib strains, these antibodies reacted with lower affinity to Ib than to Ia polysaccharide. PMID- 8406876 TI - An 88-kilodalton antigen secreted by Aspergillus fumigatus. AB - An 88-kDa component secreted in vitro by Aspergillus fumigatus has been purified by sequential chromatographic procedures. The molecule is a glycoprotein with an N-linked sugar moiety composed of mannose glucose, and galactose (16:10:1). It is recognized by antibodies from patients with aspergilloma and has potential for the immunodiagnosis of aspergilloma. The antigenicity is associated with the polypeptide part of the molecule (79 kDa). PMID- 8406877 TI - Plasmodium falciparum varies in its ability to induce tumor necrosis factor. AB - Tumor necrosis factor (TNF) has a variety of protective and pathological actions in human malaria. We report that different laboratory lines of Plasmodium falciparum which were derived from a single wild isolate (IT 4/25/5) varied widely in their ability to stimulate TNF production by human mononuclear cells. In the cloned line R29 we observed that subcultures selected for high rosetting frequency gave significantly higher levels of TNF stimulation than subcultures with low rosetting frequency, indicating that TNF induction can vary within populations that have originated from a single genotype. These results raise the possibility that the clinical severity of malaria is partly determined by the TNF inducing activity of the infecting strain of parasite. PMID- 8406878 TI - A guinea pig model for Lyme disease. AB - We report that outbred Hartley guinea pigs are susceptible to Borrelia burgdorferi. We recovered spirochetes from 57 of 60 (95%) guinea pigs inoculated when < or = 3 months of age. In contrast, animals inoculated when > or = 6 months of age were resistant to infection as defined by recovery of organisms at > or = 4 weeks postinoculation. Infection was widely disseminated: B. burgdorferi was recovered from 83% of bladders, 64% of knee joints, 57% of hearts, 48% of spleens, and 38% of spinal cords examined within 4 weeks of inoculation. Histopathologic changes were common in the heart (88%) (preferential involvement of perineural tissues near the annulus fibrosus) and bladder (76%) and were also noted in a minority of spinal cords (13%) and knee joints (9%). Western immunoblots demonstrated an immunoglobulin G response to B. burgdorferi, particularly to the 24-, 31- (OspA), 39-, and 41-kDa (flagellin) antigens. Infection was cleared from most tissues with the passage of time; spirochetes were recovered from 63% of tissues removed from guinea pigs at < or = 4 weeks after inoculation but from only 32% at > or = 8 weeks postinoculation (P < 0.001). An exception was the failure to clear spirochetes from infected knees, 90% of which were culture positive even when evaluated at > or = 8 weeks postinoculation. The guinea pig provides a new model useful for studying host spirochete interactions in Lyme disease. PMID- 8406879 TI - Virulence of capsulated and noncapsulated isolates of Pasteurella multocida and their adherence to porcine respiratory tract cells and mucus. AB - The virulence and the adherence to porcine respiratory tract cells and mucus of three toxigenic, capsular type D Pasteurella multocida isolates and their noncapsulated variants were evaluated in the present study. Loss of capsule by P. multocida, verified by transmission electron microscopy after polycationic ferritin labeling, was associated with a massive reduction in virulence of the organisms in mice. Specific-pathogen-free piglets inoculated intranasally with one of the capsulated isolates or its noncapsulated variant developed turbinate lesions characterized by bone resorption and by an inflammation of the mucosa associated with hyperplasia and squamous metaplasia of the epithelium. Infection with the capsulated isolate led to more severe lesions and atrophy of turbinates. The interactions of these P. multocida isolates with porcine respiratory tract cells and mucus were studied in vitro. The presence of capsule resulted in a decrease in binding of respiratory tract mucus were studied in vitro. The presence of capsule resulted in a decrease in binding of respiratory tract mucus to P. multocida isolates as determined by a dot blot assay. The presence of capsule also resulted in a significant decrease in adherence to porcine tracheal rings maintained in culture. The capsule seemed to mask outer membrane components which are involved in adherence. One of these components might be lipopolysaccharide since purified lipopolysaccharide bound respiratory tract mucus and blocked adherence of this microorganism to porcine tracheal rings. Our data indicate that capsular material does not seem to be involved in adherence of P. multocida to respiratory tract cells and mucus, but capsulated isolates are more virulent in mice and also in piglets. PMID- 8406880 TI - Regulation of mycobacterial growth by the hypothalamus-pituitary-adrenal axis: differential responses of Mycobacterium bovis BCG-resistant and -susceptible mice. AB - The role of the hypothalamus-pituitary-adrenal (HPA) axis in regulating the growth of Mycobacterium avium in Mycobacterium bovis BCG-resistant and susceptible congenic mice was evaluated. Restraint was used to activate the HPA axis, which resulted in an increase in the level of corticosterone in the plasma. Activation of the HPA axis increased the susceptibility of BALB/c.Bcgs mice to the growth of M. avium. In contrast, the growth of M. avium was not altered in BALB/c.Bcgr mice as a result of HPA activation. Adrenalectomy abolished the effect of HPA activation on mycobacterial growth, as did treatment of the mice with a glucocorticoid receptor antagonist, RU 486. Activation of the HPA axis also resulted in the increased susceptibility of splenic macrophages from Bcgs mice but not from Bcgr mice to M. avium growth in vitro. The production of tumor necrosis factor alpha and of reactive nitrogen intermediates by splenic macrophages from both strains of mice was suppressed as a result of HPA activation. The implications of these findings for resistance controlled by Bcg and for susceptibility to mycobacterial growth are discussed. PMID- 8406881 TI - Genetic stability and diversity of Pneumocystis carinii infecting rat colonies. AB - There is increasing molecular and antigenic evidence that Pneumocystis carinii organisms isolated from humans, ferrets, and rats are different species. In contrast, little is known about the extent of genetic diversity among P. carinii strains found within a single mammalian species. In the present study, electrophoretic karyotypes were obtained from P. carinii prepared from 10 chronically immunosuppressed rat colonies to investigate diversity at the chromosomal level. Most organism preparations produced patterns with 13 to 15 bands, but as many as 24 bands were observed in a few preparations. All bands separated between 700 and 300 kbp. Four distinct karyotype forms emerged from among the 13- to 15-band karyotypes of the 10 colonies sampled. Form 1 was shared by five rat strains from two vendors; form 2 was shared by two rat strains from the same vendor; and forms 3 and 4 were unique to their vendor colonies. Within a given rat colony, most rats harbored the same P. carinii karyotype. A survey of selected rat colonies showed that the karyotype within a vendor colony could remain stable over a period of 2 to 3 years. Hybridization of the blotted karyotypes with a repetitive DNA element isolated from rat-derived P. carinii and with single-copy gene probes showed that every chromosome in the karyotypes contained some repetitive DNA, and there was a general size concordance among the chromosomes carrying the unique gene loci. Differences in gene sequences, electrophoretic karyotypes, and hybridization profiles suggested that the immunosuppressed rats were infected by genetically distinct P. carinii strains. A provisional system of nomenclature for P. carinii that will permit differentiation of P. carinii organisms from the same mammalian host is discussed. These data show that all rats were not infected by a single type of P. carinii, that pulsed-field gradient electrophoresis can detect sufficient genetic diversity among the organism preparations to allow for characterization of the organisms, and that the genome of the organism within the rat host is relatively stable over time. PMID- 8406882 TI - Antibodies against active-site peptides common to glucosyltransferases of mutans streptococci. AB - Polyclonal antibodies were raised against peptides derived from an active-site sequence common to the family of mutans streptococcal glucosyltransferases (GTFs). The sequence contains an aspartic acid residue that functions in formation of the enzyme transition state in catalysis. Two GTFs were targeted with similar but not identical sequences in this region: one that synthesizes an alpha-1,3-linked water-insoluble glucan and a homologous GTF that synthesizes an alpha-1,6-linked water-soluble glucan. For each enzyme, an 8-mer and 22-mer peptide were prepared. The two peptide lengths were chosen in order to increase the likelihood of the peptides folding in a conformation similar to that of the native enzyme. Each peptide immunogen produced high titers of antibody in rabbits, and all antisera cross-reacted with all peptides, albeit to various degrees. Native enzyme showed weak interaction with antisera, which, on the basis of enzyme denaturation experiments, likely reflects binding to a small but finite population of denatured enzyme in the sample. GTF was assayed for inhibition in the presence of protein A-purified immunoglobulin G from each antiserum. Given the mass of the antibody and catalytic importance of the peptide, any enzyme antibody complex formation would result in enzyme inhibition. No significant inhibition was observed, which demonstrates that either polyclonal antibodies raised against each of the four peptides cannot access this active-site region, or antibodies do not recognize the native enzyme conformation. The advantages and challenges of generating antibodies against enzyme active-site peptides are discussed in the context of the crystal structure of Aspergillus oryzae alpha amylase, which has a homologous peptide segment which serves the same catalytic function. PMID- 8406883 TI - Expression of the murine interleukin-4 gene in an attenuated aroA strain of Salmonella typhimurium: persistence and immune response in BALB/c mice and susceptibility to macrophage killing. AB - Cytokines are potentially useful in vaccination as adjuvants or modulators of the type of response induced. The work below describes the expression of a cloned cytokine gene for murine interleukin-4 (mIL-4) by a live vaccine vector, an attenuated aroA strain (SL7207) of Salmonella typhimurium, in a murine model system. SL7207 was used as a carrier for two different high-level expression vectors. Both resulting strains, designated SL7207(pOmpAmIL-4) and SL7207(pKKmIL 4), expressed the cloned gene product as monitored by both immunological and biological assays. However, SL7207(pOmpAmIL-4) produced mIL-4 at higher levels and was more stable in vitro than SL7207(pKKmIL-4). When SL7207(pOmpAmIL-4) was used as a live vaccine in BALB/c mice, this strain grew and survived at higher levels than the parental attenuated strain or empty plasmid-carrying strain in spleens, livers, and intestines. This difference in growth and survival did not appear to be caused by alterations in specific lymphocyte-mediated anti Salmonella immune responses such as delayed-type hypersensitivity or serum antibody as measured by enzyme-linked immunosorbent assay; such alterations have been induced by IL-4 administration in other in vivo systems, and the lack of effect here may reflect the fact that IL-4 is not secreted from the bacteria in large quantities, most of the cytokine being in the cytoplasmic-membrane-bound fraction. Conversely, the ability of mouse macrophages to kill the bacteria in vitro was inhibited by bacterial production of mIL-4. This reduction in macrophage killing activity suggests that bacterial production of mIL-4 may be detrimental to host defense against Salmonella infection and may explain the enhanced bacterial growth and survival in vivo. PMID- 8406884 TI - Mechanism of interaction of the 85B secreted protein of Mycobacterium bovis with fibronectin. AB - The 85B protein of Mycobacterium bovis is a member of the secreted antigen 85 complex, which has been identified in a number of pathogenic mycobacteria. The 85 complex contains three components with molecular masses of 30 to 32 kDa which share the property of binding to fibronectin, a large glycoprotein present in plasma. To investigate this activity we have expressed the M. bovis 85B antigen as a recombinant protein and studied its interaction with human fibronectin. Fibronectin bound to the immobilized 85B protein in a solid-phase enzyme-linked immunosorbent assay (ELISA) and in the fluid phase in a radioimmunoassay using 125I-labelled 85B protein. In addition, fibronectin reacted with immobilized 85B in immunoblots and vice versa. Fibronectin also bound to three fragments of a cyanogen bromide digest of 85B which were subsequently identified by N-terminal sequencing. These fragments contained fibronectin-reactive peptides identified in ELISAs utilizing a set of 28 overlapping 20-mer peptides encompassing the 85B sequence. Further studies showed that the 85B protein reacted with a 32-kDa polypeptide from a limited tryptic digest of fibronectin which was identified as the collagen-binding domain. This region was confirmed as the 85B binding site by the fact that gelatin but not heparin inhibited the binding of fibronectin to 85B. These data indicate that the 85B-fibronectin interaction involves the binding of multiple regions of the 85B protein to the collagen-binding domain of fibronectin. PMID- 8406885 TI - Effect of type III group B streptococcal capsular polysaccharide on invasion of respiratory epithelial cells. AB - Group B streptococcal (GBS) capsular polysaccharide is an important virulence factor, and its role in invasion of cultured respiratory epithelial cells was investigated. A type III GBS clinical isolate, COH1, and asialo and unencapsulated isogenic transposon capsule mutants of it were compared in an in vitro invasion assay. The results demonstrated that capsule attenuated the invasion process. Invasion was not affected when the A549 epithelial cells were preincubated with purified type III GBS capsular polysaccharide. Polyclonal type III GBS capsule antibody inhibited invasion by COH1 but did not affect invasion by the capsule mutants. Serotypes Ia, Ib, Ia/c, II, and III all invaded respiratory epithelial cells but demonstrated some strain variation in magnitude of invasion. These observations led us to conclude that type III capsular polysaccharide was not essential for invasion of respiratory epithelial cells by GBS and that bacterial factors other than capsule were responsible for respiratory epithelial cell invasion. PMID- 8406886 TI - Characterization of Candida albicans cell wall antigens with monoclonal antibodies. AB - The antigenic composition of Candida albicans is very complex. In order to study the antigenic relationship between blastoconidia and germ tubes of C. albicans, we produced several monoclonal antibodies and analyzed their reactivity against cell wall antigens either in intact cells or in cells treated with dithiothreitol. Overall, four types of reactivity were found. Monoclonal antibodies 3D9 and 15C9 stained the germ tubes only when tested by indirect immunofluorescence. However, they showed a different reactivity by immunoblotting. Monoclonal antibody 3D9 reacted with antigens with molecular masses of > 200 and 180 kDa specifically expressed in the germ tube. Monoclonal antibody 15C9 reacted with antigens of 87, 50, and 34 kDa present in the germ tube extract and with antigens of 92, 50, 34, and 32 kDa present in the blastoconidium extract. The reactivity of blastoconidia treated for different times with dithiothreitol with these monoclonal antibodies was also studied by enzyme-linked immunosorbent assay. The reactivity of monoclonal antibody 3D9 did not significantly change during the cell wall extraction. However, the reactivity of monoclonal antibody 15C9 was increased for blastoconidia extracted for 60 min and decreased markedly for blastocondia extracted for 120 min. Monoclonal antibody G3B was nonreactive by indirect immunofluoresence but reacted with antigens of 47 and 38 kDa present in the germ tube extract and with an antigen of 47 kDa present in the blastoconidium extract. Monoclonal antibody B9E stained both morphological phases by indirect immunofluorescence. By immunoblotting, it reacted with antigens of > 70 kDa present in the germ tube extract and with antigens of > 63, 56, 47, and 38 kDa present in the blastoconidium extract. Based on the results presented in this study, four types of antigens are described. Type I antigens are expressed on the outermost layers of the germ tube cell wall only. Type II antigens are expressed both on the germ tube cell wall surface and within the blastoconidium cell wall. Type III antigens are found within the cell wall of both blastoconidia and germ tubes. Type IV antigens are expressed on both the blastoconidium and germ tube surface. Two types more can be hypothesized for antigens expressed on the blastoconidium cell surface and within the germ tube cell wall (type V) and for those expressed on the blastoconidium surface only (type VI). PMID- 8406887 TI - Phase variation of slime production in Staphylococcus aureus: implications in colonization and virulence. AB - Two methods commonly used for slime detection in coagulase-negative staphylococci (tube biofilm formation and colony morphology in Congo red agar) were used to study 144 ruminant mastitis Staphylococcus aureus strains. Slime production was detected in 21 strains. A majority of cells (85%) in slime-producing (SP) strains and a minority of cells (5%) in non-slime-producing (NSP) strains showed a condensed exopolysaccharide matrix (slime) surrounding the bacterial cell wall, as revealed by electron microscopy and immunofluorescence. In vivo slime production was also detected immunohistochemically after experimental infection of the mammary gland in sheep. Upon repeated subcultures in Congo red agar, NSP variants were obtained from four ovine and four bovine SP strains at a frequency ranging from 0.5 x 10(-4) to 10(-4). Because SP variants could not be obtained from NSP strains within this range or at a higher frequency, they were obtained by the tube biofilm formation (requiring repeated subculturing of NSP strains in tryptic soy broth containing 2% glucose for subsequent recovery of colonies adherent to the walls of the culture tubes). In experimental challenge, the SP variant showed a significantly higher colonization capacity than did the NSP variant of the same strain used (P < 0.001). However, the NSP variant had a higher virulence than did the SP variant (P < 0.001). These results may help to explain the different roles of S. aureus slime production cell types (SP and NSP) coexisting in disease. PMID- 8406888 TI - Association of Actinobacillus actinomycetemcomitans leukotoxin with nucleic acids on the bacterial cell surface. AB - Actinobacillus actinomycetemcomitans, a periodontopathic gram-negative bacterium, produces a leukotoxin that is a member of the RTX cytotoxin family. Although genes may function in toxin secretion, the leukotoxin is not secreted extracellularly but remains associated with the bacterial cell surface. We report here that this toxin-cell surface association is mediated by nucleic acids and directly demonstrate that the extracellular secretion of toxin occurs in growing cultures with increased ionic strength of medium. All examinations were performed with freshly harvested A. actinomycetemcomitans 301-b from anaerobic fructose limited chemostat cultures. The occurrence of cell surface-localized DNA was shown by directly digesting whole cells with the restriction endonuclease EcoRI or HindIII, which yielded many DNA fragments. The cell surface DNA constituted about 20% of the total cellular DNA. The leukotoxin was released from the whole cells by digestion with DNase I as well as restriction endonucleases. Because the leukotoxin binds ionically to DNA, it is dependent on the ionic strength of buffers or media. Accordingly, the toxin was released from cells suspended in saline at pH 7.5 in the presence of increasing amounts of MgCl2 (0 to 10 mM) or NaCl (0 to 50 mM). Moreover, a considerable quantity of leukotoxin was detected in the culture supernatant of fructose-limited chemostat cultures when sodium succinate solution was pumped into the steady state as an additional salt (30 and then 50 mM). This toxin-DNA association was also found in well-characterized strains including not only the leukotoxin-producing ATCC 29522 but also the toxin production-variable ATCC 29523 and the non-leukotoxin-producing ATCC 33384 when these strains were grown in the chemostat culture. PMID- 8406889 TI - Roles of leukotriene B4, prostaglandin E2, and cyclic AMP in Campylobacter jejuni induced intestinal fluid secretion. AB - Infection of rabbit ileal loops with inflammatory Campylobacter jejuni strains caused elevation of cyclic AMP, prostaglandin E2, and leukotriene B4 levels in tissue and fluids. Incubation of cultured Caco-2 cells with loop fluids caused elevated cellular cyclic AMP levels, an effect which was inhibited by antiserum against prostaglandin E2. PMID- 8406890 TI - Microtubule inhibitors block Cryptosporidium parvum infection of a human enterocyte cell line. AB - No effective therapy exists for Cryptosporidium parvum, a coccidial protozoan parasite that causes severe diarrhea in patients with AIDS. The role of microtubules in parasite invasion of host cells was investigated by incubating 10(7) oocysts with a HT 29.74 cell line for 24 h in the presence of microtubule disrupting drugs. The number of parasites per 1,000 cells was reduced by 77% (P < 0.001, n = 4) from 182 +/- 3 in untreated cells to 42 +/- 4 in cells treated with 10(-4) M colchicine. Inhibition of C. parvum infection was concentration dependent. Similar results were seen with a second microtubular depolymerization agent, vinblastine. These data suggest that microtubules are important in host cell invasion by C. parvum and may represent targets for development of new therapeutic drugs for treatment of cryptosporidiosis. PMID- 8406891 TI - Effect of mannose-binding protein on binding of Cryptococcus neoformans to human phagocytes. AB - The capacity of human peripheral blood monocytes, neutrophils, and monocyte derived macrophages to bind the yeast Cryptococcus neoformans was increased when the phagocytes were cultured on surfaces containing recombinant human mannose binding protein. In contrast, soluble mannose-binding protein had no effect on cryptococcal binding by phagocytes. PMID- 8406892 TI - Virulence determinants in nontoxinogenic Escherichia coli O157 strains that cause infantile diarrhea. AB - Ten sorbitol-fermenting Escherichia coli O157 strains that cause infantile diarrhea and are positive in the fluorescence actin staining test were determined to be negative for Shiga-like toxin (SLT) genes. We amplified their complete eae genes, contrasting them with those of SLT-producing E. coli O157 by restriction fragment length polymorphism analysis and nucleotide sequence analysis of a 400 bp stretch of the 3' end of eae. The data substantiated the presence of two eae genotypes within serogroup O157, one resembling eae of enteropathogenic E. coli (EPEC) strain E2348/69, found in nontoxinogenic E. coli O157 strains, and the other resembling eae of EHEC strain EDL 933, found in toxinogenic E. coli O157 strains. Another EPEC-specific virulence determinant was also shown to be large plasmids harboring EPEC adherence factor sequences. The SLT-negative E. coli O157 strains described here fall under the heading of EPEC, which serves as an explanation for their virulence in infants, and represent a third pathogroup within serogroup O157. PMID- 8406893 TI - Endogenous gamma interferon mediates resistance to Brucella abortus infection. AB - Depletion of endogenous gamma interferon (IFN-gamma) with anti-IFN-gamma monoclonal antibody resulted in increased numbers of Brucella abortus in the spleen and liver of infected CBA mice. This increase was accompanied by a decrease in splenomegaly and a lower proportion of macrophages in the spleen. Furthermore, treatment of recipient mice with anti-IFN-gamma antibody blocked the adoptive transfer of resistance with immune T cells. Together, the results indicated that endogenous IFN-gamma plays an important role in mediating resistance to primary and secondary Brucella infection. PMID- 8406894 TI - Effect of orally administered monoclonal antibody on persistent Cryptosporidium parvum infection in scid mice. AB - Scid mice, persistently infected after exposure to 10(7) Cryptosporidium parvum oocysts, were treated daily for 14 to 17 days with 0.4 mg of monoclonal antibody (mAb) 17.41 administered by the oral route. Mice receiving mAb 17.41 shed significantly fewer (P < 0.005) C. parvum oocysts than scid mice receiving isotype control mAb. Intestinal (but not gastric) infectivity scores were also reduced for scid mice treated with mAb 17.41 (P < 0.01). PMID- 8406895 TI - Isolation and nucleotide sequence of the gene encoding cytotoxic necrotizing factor 1 of Escherichia coli. AB - Cytotoxic necrotizing factors (CNFs) are dermonecrotic protein toxins produced by human and animal clinical isolates of Escherichia coli. In this study, the CNF1 determinant was isolated and sequenced, showing that expression of biologically active toxin is governed by a unique open reading frame encoding a protein of 1,014 amino acids with a predicted molecular mass of 113.7 kDa. Nucleotide and protein data base searches showed significant homology between CNF1 and the dermonecrotic toxin of Pasteurella multocida. In particular, the two toxins were found to share a hydrophobic region of about 220 amino acids which is a potential membrane-spanning domain. PMID- 8406896 TI - The 39-kilodalton outer membrane protein of Proteus mirabilis is an OmpA protein and mitogen for murine B lymphocytes. AB - Partial amino acid sequence analysis of a major outer membrane protein of Proteus mirabilis (39-kDa protein) indicates that it is an OmpA protein. The mitogenic activities of the 39-kDa protein for murine lymphocytes were also investigated with T lymphocytes isolated by passing spleen cells over columns of nylon wool fiber and B lymphocytes obtained by treating spleen cells with monoclonal antibodies to Thy1 plus complement. The 39-kDa protein showed little activity in stimulating T cells to proliferate but was strongly mitogenic for B cells. PMID- 8406897 TI - Effect of cholera toxin on vaccine-induced immunity and infection in murine schistosomiasis mansoni. AB - Intradermal vaccination of mice with soluble adult worm antigen (SWAP) in combination with Mycobacterium bovis BCG (Swedish strain) induced significant protection against subsequent infection with Schistosoma mansoni cercariae. When cholera toxin (CT) was used as an adjuvant in combination with SWAP or fraction A, no significant protection was observed. However, intradermal vaccination in combination with CT triggered a strong anti-SWAP antibody response and induced a strong delayed-type hypersensitivity response to schistosome antigens (SWAP or fraction A), one significantly higher than that in the SWAP-BCG group. In addition, vaccinating mice intranasally with SWAP or cercarial antigen together with CT as adjuvant failed to induce any significant protection. Surprisingly, mice given CT alone intranasally revealed a significantly enhanced worm burden. These findings suggest that mucosal application of CT may modulate the host parasite relationship in favor of parasite survival. PMID- 8406898 TI - An Fc gamma RII-, Fc gamma RIII-specific monoclonal antibody (2.4G2) decreases acute Trypanosoma cruzi infection in mice. AB - In order to study the role of Fc gamma Rs in Trypanosoma cruzi infection in mice, the 2.4G2 monoclonal antibody (MAb), specific to the extracellular domains of Fc gamma RII and Fc gamma RIII, was injected intraperitoneally into mice. Flow cytometry studies of uninfected mice showed that 2.4G2 MAb bound to peritoneal and lymph node cells, respectively, on days 2 and 6 after injection. Repeating 2.4G2 injections every 3 to 4 days decreased the availability of Fc gamma Rs on peritoneal, lymph node, and spleen cells. Injections of 2.4G2 MAb into T. cruzi infected mice, at days -1, 3, 7, 11, 16, 20, and 24 relative to infection, reduced mortality in comparison with that in infected animals injected with an unrelated MAb (50 versus 93.3% mortality; P < 0.01). Parasitemia in 2.4G2-treated mice was significantly (three times) lower than in control animals on days 21 and 24 postinfection (P < 0.05), before parasite-specific antibodies were detectable at significant levels. Immunoglobulin and T. cruzi-specific antibody levels were similar in all groups of mice. These results indicate that repeated injections of 2.4G2 MAb administered to acutely infected mice reduce the in vivo infection level, suggesting that Fc gamma Rs play a role in the early host invasion by T. cruzi parasites. PMID- 8406899 TI - Evidence that extracellular components function in adherence of Actinobacillus actinomycetemcomitans to epithelial cells. AB - Extracellular microvesicles and a highly proteinaceous polymer associated with a leukotoxin-producing strain, Actinobacillus actinomycetemcomitans SUNY 75, were shown to increase adherence of other weakly adherent A. actinomycetemcomitans strains to KB epithelial cells. PMID- 8406900 TI - Interleukin 10 protects mice against staphylococcal enterotoxin B-induced lethal shock. AB - We investigated the ability of interleukin 10 (IL-10) to protect mice against lethal shock induced by staphylococcal enterotoxin B (SEB). Treatment of mice with IL-10 prevented the death of mice injected with SEB in a dose-dependent manner. IL-10-mediated protection was apparent when administered either prior to or concurrent with SEB but was less effective when administered following SEB injection. This observation indicates that IL-10 is capable of regulating T-cell activation in vivo. PMID- 8406901 TI - Biological monitoring of acrylonitrile exposure through a new analytical approach to hemoglobin and plasma protein adducts and urinary metabolites in rats and humans. AB - A new, simple and fast procedure of measuring acrylonitrile (ACN) in ACN derived mercapturic acids such as S-(2-cyanoethyl)-L-cysteine(CyEC), and in hemoglobin (Hb) and plasma protein adducts and urinary metabolites in rats and humans exposed to ACN was developed. ACN in mercapturic acids or proteins was analyzed by capillary gas chromatography (GC) by liberating ACN at a high-temperature in the injector port of GC with or without oxidizing sulfur atoms of the ACN-bound cysteines into sulfoxide form by hydrogen peroxide in vitro. At 350 degrees C, more than 90% of ACN in authentic CyEC was recovered by this method. Increasing a single ip dose of ACN from 5 to 50 mg/kg produced proportional increases in ACN bound to Hb 24 hr after the treatment. The alkylation of plasma protein with ACN was about 1/10 as low as that of Hb. After repeated daily ip doses of 1-10 mg/kg, ACN in Hb decreased with a half-life of about 9 days. ACN was also detected in the blood of workers exposed to ACN for 1 to 10 years at a Siberian synthetic rubber factory. PMID- 8406902 TI - Biological monitoring of workers exposed to styrene and acetone. AB - Twenty-two workers exposed to styrene and acetone in two fiberglass industries were monitored on Monday and Thursday for 8 hours using passive dosimeters. Urine samples were collected at the end of the workshift and before the start of the work on the next morning (Tuesday and Friday). The charcoal disks of the passive dosimeters were analysed by gas-chromatography. Mandelic acid (MA) and phenylglyoxylic acid (PGA) were measured using a HPLC method; values were expressed in mg/g of creatinine. The 8-h TWA exposure values for styrene and acetone ranged respectively from 22 to 522 mg/m3 and 40-1581 mg/m3 on Monday; 25 423 mg/m3 and 55-579 mg/m3 on Thursday. Styrene TWA exposure values significantly correlate with the sum of metabolites at the end of workday (r = 0.70 on Monday and r = 0.95 on Thursday) and also at the next morning (r = 0.86 on Tuesday and r = 0.85 on Friday). A styrene exposure level of 213 mg/m3 (ACGIH-TLV) was associated with an excretion of metabolites (MA+PGA) higher on Thursday (803 mg/g creat) than on Monday (570 mg/g creat). The same result was found on Friday (459 mg/g creat) compared with Tuesday (305 mg/g creat). Moreover our data show that the simultaneous exposure to acetone does not modify the excretion of MA. In conclusion the TLV of styrene is associated with different values of metabolites at the beginning and at the end of the work-week. PMID- 8406903 TI - Practical approaches to the assessment of work-related risks. AB - The control of work-related risks calls for practical improvements in job content and the working environment. For assessment of risks, it is essential to resort to the most practical methods in the local context. Important common methods are examination of the process, equipment and organisation of work, walk-through surveys, evaluation of risk factors in the working environment, inquiries and questionnaires as well as monitoring of various health indicators. Biological tests and other health indicators can thus only be regarded as components of overall occupational health-risk assessment strategy, with their advantages, disadvantages and limitations. Important are the validity of biological tests and the ethical aspects related to their use, such as confidentiality of data and the individual worker's informed consent, preference being given to noninvasive methods. Occupational health services should thus play a positive role by providing practical advice, training and information for participatory risk assessment keeping in view such an overall strategy that facilitates immediate preventive action. Biological monitoring should be applied when the additional guidance it provides on preventive action is essential. Proposals to use biological monitoring must emphasise this justification aspect. PMID- 8406904 TI - Biological monitoring of hexachloroethane. AB - A small group (n = 12) of military white smoke munition workers provided blood plasma during a production break (S I) and after five weeks' production (S II) of a hexachloroethane (HCE)/titanium dioxide formula. Plasma was also obtained from a sex and age matched control group (n = 12) and a group (n = 13) of previously HCE-exposed workers, respectively. HCE in plasma (P-HCE) was determined with gas chromatography and electron capture detection. No HCE was found in the plasma samples from the two control groups. In the HCE exposed group the mean (+/- SD) P HCE level increased almost two orders of magnitude from S I (0.08 +/- 0.14 microgram/l) to S II (7.30 +/- 6.04 micrograms/l) despite efforts to minimize the internal dose. Biological monitoring of HCE could be useful in occupational hygiene. PMID- 8406905 TI - Biological monitoring for occupational exposures to o-toluidine and aniline. AB - Epidemiological evidence that occupational exposure to o-toluidine and aniline is associated with an increased risk of bladder cancer led to efforts to identify biomarkers of workplace exposures to these aromatic amines. For the determination of o-toluidine and aniline in worker urine specimens, a method using high performance liquid chromatography (HPLC) followed by electrochemical detection was developed. The limits of detection were 0.6 microgram/l and 1.4 micrograms/l for o-toluidine and aniline, respectively. Recovery of o-toluidine and aniline from spiked urine averaged 86% and 93%, respectively, over a range of 4-100 micrograms/l. Reproducibility in the range 2-100 micrograms/l for analyses of split field samples was 13% (average RSD) for o-toluidine and 16% (average RSD) for aniline. Application of this method to pre- and post-shift samples collected from potentially exposed and unexposed workers indicated elevated concentrations of o-toluidine and aniline in urine from exposed workers. To develop methods for biomarkers of internal dose, o-toluidine binding to the blood proteins hemoglobin and albumin was investigated utilizing in-vivo (rodent) and in-vitro (hemoglobin and albumin) studies. Base-hydrolyzable protein adducts were analyzed by HPLC (fluorescence) and/or GC/electron capture (EC). The methods were compared for sample preparation requirements, selectivity and sensitivity. While the GC/EC method was more sensitive than HPLC, the presence of interfering peaks limited the utility of this approach. Results from these studies suggested that the HPLC method could be useful for determination of o-toluidine exposures in individuals acutely or chronically exposed to high levels. PMID- 8406906 TI - Assessment of exposure to chloramphenicol and azathioprine among workers in a South African pharmaceutical plant. AB - There have been very few published studies that have evaluated exposure to myelotoxic drugs among production workers in pharmaceutical plants. Previous studies have focussed mainly on nurses and evaluated exposure to cytotoxic drugs using urine mutagenicity as a marker of exposure. The aim of this study was to evaluate the exposure of workers involved in the production of chloramphenicol and azathioprine. Exposure was evaluated utilising biological monitoring, biological effect monitoring and environmental monitoring. Biological monitoring included plasma chloramphenicol levels, plasma 6-mercaptopurine and urine 6 thiouric acid levels. These were analysed using high performance liquid chromatography. Myelotoxic effect was assessed by measuring the haematological indices of bone marrow function. The exposed 17 workers were compared to matched controls of equal numbers. Neither substance could be detected in serum nor urine by the analytical methods employed. However, haematological indices demonstrated a significantly decreased mean reticulocyte and neutrophil count in the azathioprine exposed group. Industrial hygiene measurements demonstrated contamination of the air inside the airhood of exposed workers. In conclusion, it is evident that workers involved in the production of both these drugs are at risk of developing adverse health effects. Furthermore, more sensitive analytical methods need to be developed to evaluate absorption of myelotoxic chemicals among occupationally exposed workers. PMID- 8406907 TI - Comparative evaluation of blood and urine analysis as a tool for biological monitoring of n-hexane and toluene. AB - Blood and urine samples were collected from 57 male Japanese solvent workers [exposed to n-hexane (Hex-A), ethyl acetate, and toluene (Tol-A) at 1.5, 2.3, and 2.3 ppm as GM-TWA, respectively] and also from 20 male nonexposed workers at the end of a 8-h shift, and analyzed for n-hexane (Hex-B) and toluene (Tol-B) in blood, and n-hexane (Hex-U), toluene (Tol-U), 2,5-hexanedione [both with (HD U/cHYD) and without hydrolysis (HD-U/sHYD)] and hippuric acid (HA-U) in urine. Regression analysis showed that both Hex-B and Tol-B correlated significantly with corresponding exposure to the solvents. Solvents in urine (Hex-U and Tol-U) also correlated with solvents in air but with smaller correlation coefficients than the solvents in blood. Both HD-U/cHYD and HD-U/sHYD showed significant correlation with Hex-A, but HA-U failed to do so with Tol-A. Based on the correlation among biological exposure indicators and solvent concentration in air, sensitivity as an exposure indicator was compared between the solvent in blood and the metabolite in urine in terms of the lowest solvent concentration at which the exposed can be separated (with statistical significance) from the nonexposed (the lowest separation concentration; LSC). The LSC was 3.9 ppm for Hex-B, 1 to 2 ppm for HD-U/sHYD and 10 to 30 ppm for HD-U/cHYD, suggesting that HD-U/sHYD is superior even to Hex-B in detecting low n-hexane exposure; this high sensitivity of HD-U/sHYD is due to the absence of HD-U/sHYD in the urine from the nonexposed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406908 TI - Immunochemical assessment of the influence of nutritional, physiological and environmental factors on the metabolism of toluene. AB - Factors influencing the metabolism of toluene were investigated in rats using monoclonal antibody (MAb) to cytochrome P450 (P450). At low toluene concentrations, P450 IIE1 was primarily involved in the metabolism of toluene, whereas P450 IIC11/6 was involved at high concentrations. A low-carbohydrate diet induced P450 IIE1 and resulted in an increase in toluene metabolism. The intake of fat did not influence the metabolism. A lowered protein intake decreased not only the total content of P450 but also the P450 IIC11/6. Fasting and ethanol consumption also enhanced toluene metabolism via the induction of P450 IIE1. The metabolic rate of toluene in adult male rats was 4-fold higher than in immature males and adult females at a high substrate concentration because of the high level of P450 IIC11/6 in adult males, whereas no difference was noted between adult and immature females. Although development did not influence toluene metabolism in males at a low substrate concentration, the metabolic rate in adult female rats was significantly lower than that of immature females and males; this may be due to the decrease in P450 IE1 with development. Diabetic status influenced toluene metabolism in rats by affecting several kinds of P450 isozymes. Toluene exposure also affected its own metabolism by increasing P450 IIE1 and P450 IIB 1/2, and decreasing P450 IIC11/6. A significant difference in toluene metabolism was observed among rat, mouse and human liver microsomes. Thus, when considering the factors affecting toluene metabolism, it is important to elucidate the change in specific P450 isozyme composition related to the modifications, and their affinities to toluene. PMID- 8406909 TI - Biological monitoring of toluene and DNA polymorphism in humans. AB - The effects of genetic polymorphism in aldehyde dehydrogenase 2 (ALDH2) on alcohol intakes and toluene metabolism were studied in workers who handle toluene. The genotypes of ALDH2 were divided into three groups, which were the homozygous genotype of normal ALDH2 (NN), the homozygous genotype of an inactive one (DD) and the heterozygous genotype of normal and inactive ones (ND), using the polymerase chain reaction (PCR) method in a solution containing ddGTP. The alcohol intake of the DD group was very small. On the other hand, the alcohol intake of NN group was higher than that of DD group. That of ND group was about half way between those of NN group and DD group. The alteration of toluene metabolism by the polymorphism of ALDH2 was not clear because the exposure levels of the workers to toluene were low. PMID- 8406910 TI - A descriptive and mechanistic study of the interaction between toluene and xylene in humans. AB - This study was undertaken to characterize the mechanism of toxicokinetic interaction between toluene (TOL) and m-xylene (XYL) in the rat using physiologically-based toxicokinetic (PBTK) modeling approach. First, the metabolic rate constants were determined by conducting closed-chamber inhalation exposures with individual solvents (Vmax: TOL = 4.8, XYL = 8.4 mg/hr/kg; Km: TOL = 0.55, XYL = 0.2 mg/l). Then, using the same experimental set-up, rats were exposed to different binary mixtures of TOL and XYL. PBTK analysis of the data showed competitive inhibition as the plausible mechanism of TOL/XYL interaction. This mechanistic modeling study suggests that the interaction between TOL and XYL is likely to be observed when the exposure concentration exceeds 50 ppm of each solvent. PMID- 8406911 TI - Evaluation of stable isotope-labeled probes in the study of solvent pharmacokinetics in human subjects. AB - The relationship between biomarkers of exposure (such as concentrations of toxicants in blood or breath, or metabolites in urine) and toxicant dose for individuals is influenced by many person- and episode-specific factors which contribute to overall variability in biomarker level for a given dose. This variability results in imprecise biological marker-based estimates of dose for individuals. We hypothesize that pharmacokinetic data from stable-isotope (deuterated) analogs can be used with a pharmacokinetic model to account for individual-related sources of variation, leading to more precise methods of dose estimation for individuals. To establish the degree of similarity in the pharmacokinetics of unlabeled (d0-) and fully deuterated (D8-) toluene, 21 men (ages 20-45) inhaled an equal molar mixture for 2h. Washout kinetics for both compounds were followed for 4 d in alveolar air and blood. Both compounds exhibited three-phase elimination kinetics in both fluids. The third phase was not always definable for d0-toluene because of concurrent uncontrolled environmental exposures. Considering data from only the first two phases, concentrations of d0- and d8-toluene in alveolar air and blood were well correlated for all subjects, even though pharmacokinetic parameters varied among individuals by 5-9 folds. Further experiments are needed to discern whether correlations between d0- and d8-toluene for the third phase are influenced by an isotope effect; present data support use of d8-toluene as a suitable probe for d0 kinetics. PMID- 8406912 TI - Correlation between PAH airborne concentration and PAH-DNA adducts levels in coke oven workers. AB - In order to evaluate the correlation between peripheral blood leukocyte DNA adducts as an indicator of exposure to polycyclic aromatic hydrocarbons (PAH) and the airborne contamination of PAH at the work places, a survey of a cohort of coke-oven workers has been carried out. In each workplace, total and specific PAH airborne concentrations were measured. Among the job title groups, the highest proportion of subjects with levels of adducts above the detection limit and the highest mean value were observed in the door-operators. The correlation between median values of environmental benzo(a)pyrene concentration and mean values of adducts concentration support the fact that the maintenance workers group has a higher relative risk (RR) to have detectable level of PAH-adducts, which is estimated to be 1.84 for an increase of 1 micrograms/m3 of benzo(a)pyrene. PMID- 8406913 TI - Development of an immunoassay to detect hemoglobin adducts formed by benzene exposure. AB - Polyclonal murine antibodies that recognize the adducts formed by benzene metabolites in hemoglobin (Hb) were prepared and used to develop immunoassays. In competitive inhibition assays, the concentration of competitor needed to reduce the signal by 50% (IC50) was less than 10 pmoles for hydroquinone-hemoglobin (HQHb) adducts and less than 1 pmole for 1,2,4 trihydroxybenzene-hemoglobin (TriOH Hb). Hemoglobin (Hb) incubated with either phenol or catechol (CAT) did not elicit antibodies suitable for quantitative immunoassays. The metabolite directed immunoassays were tested using hemoglobin from mice previously administered [C14] benzene for two to four weeks. The most sensitive assay for hydroquinone measured 0.49 pmoles adduct/40 pmoles Hb (191 pmoles adduct/mg Hb) in mice treated with 200 mg/kg benzene (P < 0.05, Student's t test). TriOH Hb adducts were not detected. PMID- 8406914 TI - Japanese experience in biological monitoring. AB - In Japan, industrial health control is accomplished by control of the working environment, work practice management and health care. Periodical biomonitoring of workers exposed to lead and 8 popular organic solvents, including mixed solvents, became mandatory as of October 1, 1989, with an ordinance issued by the Ministry of Labor. This ordinance states that the results of measurements of lead and the 8 solvents in each work place must be classified into 3 categories, named distributions 1, 2 and 3 based on the biological level of the determinant, and then must be reported to the Labor Standards Inspection Office. Distribution 3 encompasses workers having concentration levels above the BEIs of ACGIH except for the concentration of lead in the blood. The National Federation of Industrial Health Organizations surveyed the state of biological monitoring in Japan by a questionnaire to the laboratories. The total number of cases examined in the fiscal year 1990 was about 110,000 for the biomonitoring of lead, and about 520,000 for the monitoring of urinary metabolites of the 8 organic solvents. The ratio of the number of workers in distribution 3 to that of the total workers examined was 0.4% for urinary delta aminolevulinic acid and 0.2-3.5% for the urinary metabolites of each organic solvent. Similar results were obtained from 7 major laboratories. To achieve large-scale biological monitoring in future, a method is necessary for evaluating the exposure or health risks of workers exposed to mixed organic solvents by estimating their urinary metabolites. PMID- 8406915 TI - Determination of butoxyacetic acid and N-butoxyacetyl-glutamine in urine of lacquerers exposed to 2-butoxyethanol. AB - To determine the fraction of butoxyacetic acid (BAA) which is excreted as the amino acid conjugate N-butoxyacetylglutamine (BAA-GLN), urine samples of six lacquerers exposed to 2-butoxyethanol (BE) were collected before and after work and analysed using an HPLC-method which allows the simultaneous quantification of both BAA species. Whereas the pre-shift samples contained only little or no butoxyethanol-related material, concentrations of BAA and BAA-GLN amounted collectively to up to 7 mmol/l in the samples obtained at the end of work. The ratio BAA-GLN vs. total BAA ranged from 0.16 to 0.64 (mean value 0.48) indicating that a substantial fraction of BAA was eliminated as the amino acid conjugate. The results demonstrate that BAA-GLN is an important metabolite of BE in man. Procedures employed for the biological monitoring of exposure to BE should therefore include the quantification of BAA-GLN, otherwise exposure levels would be underestimated. PMID- 8406916 TI - Biomarkers of individual susceptibility to carcinogens: application for biological monitoring. AB - In order to develop new markers of individual susceptibility to various human carcinogens, we studied some parameters of formation and metabolism of carcinogens, as well as DNA adducts formation and DNA repair in animals and humans. Following an i.p. administration of benzo(a)pyrene (BP) to the rats, levels of urinary excretion of BP-7,8-diol correlated with tumour latency. A high correlation was found between excretion of this metabolite and BP-DNA adducts level in the liver. Healthy smokers excreted higher quantities of BP-7,8-diol, than smoking lung cancer patients, thus confirming the suggestion on existence of cancer-prone phenotype. N-nitroso compounds formed most efficiently in stomach juice of children with superficial gastritis who therefore could be at high risk of stomach cancer. N-ethyl-N'-nitro-N-nitrosoguanidine induced stomach cancer earlier in monkeys with a low level of DNA repair enzyme, O6-alkylguanine-DNA alkyltransferase (AGT) in gastric mucosa. Overall, these markers can be helpful in predicting individual susceptibility to carcinogens. PMID- 8406917 TI - Evaluation of biological monitoring parameters for occupational exposure to toluene. AB - A survey was conducted in a rotogravure printing plant with inhalatory and percutaneous exposure to toluene. Workers (n = 9) were followed for 2 consecutive days and the frequency and duration of skin contact with toluene were monitored. In order to assess percutaneous absorption an airstream helmet was worn during one day. Urine and exhaled air samples were collected simultaneously 5 times each day for toluene (urine and breath) and hippuric acid (urine). The mean (personal air sampling) exposure concentration was between 30 mg/m3 and 600 mg/m3. The best biological monitoring parameter of external exposure (without a helmet) was the concentration toluene in exhaled air 8 h after work (r = 0.99). While wearing the airstream helmet the relationship between external exposure (measured in the helmet) and concentrations in exhaled air and urine deviated from the preceding relations. This was likely the result of the high body burden and not of skin contact with toluene. Skin contact with toluene (usually by cleaning of the hands) was limited to 0-30 minutes a day, with an average of about 5 minutes. During experimental exposure (n = 6) in which the hands were washed with toluene for 5 minutes the toluene in exhaled air (max after 1040 min) clearly demonstrated skin absorption of toluene. The next morning 0.1 mg/m3 toluene was still detectable; this was less than the concentration measured the next morning in exhaled air of workers: between 0.5 and 10 mg/m3. PMID- 8406918 TI - Assessment of exposure to trichloroethylene and tetrachloroethylene in the population of Zagreb, Croatia. AB - Human exposure to trichloroethylene (TRI) and tetrachloroethylene (perchloroethylene, PER) through air, food and drinking water was assessed by biological monitoring. By stratified sampling 39 subjects with no known solvent exposure were selected from the residents of the city of Zagreb (the capital of the Republic of Croatia). TRI and PER were determined in blood and their metabolites trichloroethanol (TCE) and trichloroacetic acid (TCA) in plasma and urine. Drinking water samples were also taken for TRI and PER determination. TRI was present in measurable concentrations in 22 subjects (range: 0.026-0.090 micrograms/l) and PER in 31 subjects (range: 0.031-2.540 micrograms/l). TCE was not detected in any of the samples, while TCA was determined in all plasma (range: 13.54-160.42 micrograms/l) and urine samples (range: 1.64-291.76 micrograms/24h). In drinking water samples TRI and PER values were also in a very wide range: 0.69-35.90 micrograms/l and 0.36-7.80 micrograms/l, respectively. The variation of all results presented is most probably a reflection of the different environmental contamination with TRI and PER in the different city areas. PMID- 8406919 TI - Urinary metabolite levels and symptoms in Filipino workers using organic solvents. AB - To compare symptoms with urinary metabolite levels, 900 workers from 7 organic solvent-using industries were studied. Urinary metabolites were determined using a high performance liquid chromatograph. Urinary hippuric acid concentrations exceeding the reference value (2.5 g/g creatinine) were found in 78 (8.7%) workers. However, only 3 (0.3%) and 1 (0.1%) of the participants exceeded the reference value for mandelic (0.8 g/g creatinine) and total methylhippuric acid (1.5 g/g creatinine), respectively. The sum of the values of the ratio of measured urinary metabolite concentration to the corresponding ACGIH's biological exposure indices (BEI) [(HA/BEI of HA + MHA/BEI of MHA + MA/BEI of MA)] exceeded 1.0 in 166 (18.4%) workers. Majority of them were from the footwear manufacturing industry (63/129 or 49.2%). Questionnaire interviews were also administered to determine the prevalence of symptoms while at work (acute symptoms) or within the past 6 months (chronic symptoms). Urinary metabolite levels of individual and mixed solvents were compared with the symptoms of all workers. Analysis using Spearman's rank correlation showed in workers whose urinary hippuric acid exceeded 3.75 g/g creatine (1.5 x BEI), significant correlation between their hippuric acid levels and subjective complaints. Workers whose sum of the values of the ratio of measured urinary metabolite concentration to corresponding BEI exceeded 1.5 were selected and comparing this level with their symptoms, significant correlation was also noted in some complaints. PMID- 8406920 TI - On the need of a sampling strategy in biological monitoring: the example of hexane exposure. AB - Ambient and biological monitoring of hexane exposure were repeatedly carried out in 14 female shoe makers. Airborne hexane (Ci-H) was measured in 4-h samples collected by a diffusive method. Urinary spot samples were collected before, during (at noon), and at the end of a work shift. 2,5-Hexanedione (2,5HD) in urine collected at noon was poorly related to morning Ci-H. End-of-shift 2,5HD were also poorly related to afternoon air samples. The correlation was still relatively low when end-of-shift 2,5HD was related to 8-h TWA Ci-H (r = 0.44; P < 0.01 on a linear scale, and r = 0.58, P < 0.01 on a log-log scale). End-of-shift 2,5HD levels estimated on the basis of pre-shift values using a mathematical model were much higher (2.3 times on average) than those experimentally measured during the study period. Owing to its relatively long half-time, 2,5HD seems to be influenced not only by current exposure, but also by hexane absorbed during the day(s) preceding sampling. The lack of a sampling strategy may account not only for inconsistencies between environmental and biological data, but also for a possible misuse of biological monitoring when utilized for risk assessment. Despite sometimes poor correlations with Ci-H, 2,5HD may still be preferred to other indicators as a marker of effective internal dose. A sampling strategy should ensure that measured values are representative of the individual risk for adverse effects. PMID- 8406921 TI - Biological monitoring of workers occupationally exposed to carbon disulphide in a rayon plant in Brazil: validity of 2-thiothiazolidine-4-carboxylic acid (TTCA) in urine samples taken in different times, during and after the real exposure period. AB - Fifteen workers from a rayon plant in Brazil were monitored. Air samples were taken during a mean period of 5.8 hours out of an 8 hour workshift, in three different adsorbing tubes. Five urine samples were taken at 4 hour interval from the beginning of the shift, and at the beginning of the next shift in which 2 thiothiazolidine-4-carboxylic acid (TTCA) concentration was analysed. Data from seventeen "individuals" were statistically studied, with the aim of establishing the best and the most practical sampling strategy of biological monitoring for workers occupationally exposed to CS2. For those chronically exposed to CS2, TWA air concentration lower than 30 and higher than 10 mg/m3, it was found that the higher the exposure levels, the lower was TTCA concentration in the end of shift urine samples. The results may be explained by dietary habits and/or by the small number of examined population or even eventual casualty. On the other hand they raise other hypothesis involving the chronicity of exposure which may lead to important changes in metabolism influencing the excretion rate of TTCA. PMID- 8406922 TI - The effect of respiratory protection with biological monitoring on the health management of lead workers in a storage battery industry. AB - Respiratory protection with maintenance free respirator and regular biological monitoring with the measurement of zinc protoporphyrin for one year period not only made significant decreases of biological indices indicative of lead exposure (blood lead, delta-aminolevulinic acid in urine) in a storage battery workers, but also reduced the prevalence of lead related symptoms. PMID- 8406923 TI - Biomonitoring of workers exposed to pesticides. AB - The potential clastogenic effects of pesticides was investigated in 71 floriculturists exposed to complex chemical mixtures. Exposed and referent subjects were selected from the same geographical area located in north western Italy. A significant association between micronuclei frequency and occupational exposure to pesticides was found (RR = 1.25). A positive dose-response gradient was observed with years of employment (used as an index of cumulative exposure) as a floriculturist. Individuals working exclusively in greenhouses (confined spaces) showed higher micronuclei levels than subjects working in open fields. The study supports the hypothesis that human exposure to pesticides causes a clastogenic damage. PMID- 8406924 TI - Biological monitoring for pesticide exposure--the role of human volunteer studies. AB - Predictions of human pesticide metabolism which are needed for the interpretation of biological monitoring data are frequently made from animal studies. Consequently, assumptions have to be made about the relationship between absorbed dose and metabolite excretion. The results from two human volunteer studies highlight the problems associated with extrapolating from animal studies in this way. The pyrethroid insecticide cypermethrin shows markedly different metabolite patterns when administered orally or dermally in man. Following dermal dosing the ratio of trans/cis cyclopropane acids is approximately 1:1, compared to 2:1 after oral administration. The ratio of total cyclopropane acids to phenoxybenzoic acids also differs depending on the route (dermal 1:4, oral 1:0.8). A knowledge of human metabolism by these two routes enables a much more meaningful interpretation of biological monitoring measurements. The herbicide molinate forms a mercapturate conjugate as a major urinary metabolite in the rat (35%). In volunteers at low dose levels this metabolite is present at insignificant levels (< 1%) and 4-hydroxymolinate is a much more abundant metabolite (39%). This shows that extrapolation between species can be very misleading. It is concluded that the benefits of using human volunteers for metabolism studies at low doses far outweigh the minimal risks involved. As a basis for biological monitoring such studies can lead to a greatly improved risk assessment for pesticides in use. PMID- 8406925 TI - Biomonitoring of nickel and chromium in human pulmonary tissue. AB - Nickel (Ni) and chromium (Cr) and some of its compounds may be able to induce cancer in the lungs as well as in the nose and paranasal sinuses after occupational exposure. Latency periods amount to 20 years and more. Therefore objective exposure data are not available in the most cases and expert evaluation of the causal connection is often difficult. Recent investigations have shown, that Ni and Cr can cumulate in human lung tissue after occupational exposure. For the evaluation of "normal" Ni- and Cr-values a total of 495 human lung tissue samples of 30 occupationally non-exposed persons were analysed by AAS including ZEEMAN-compensation after wet oxidative digestion. Additional samples of 10 deceased persons who have been occupationally exposed to nickel in previous times by nickel-refining and welding, especially flame spraying have been investigated. The median Ni- and Cr- concentrations in the lungs of the non-exposed persons ranged between 20-40 resp. 133-277 ng/g (wet weight). In nickel refinery workers Ni- concentrations were found which exceeded the normal range about 1,000. In welders, especially flame sprayers, also values more than 100 times higher could be analysed for Ni and Cr. Partially these concentrations were found years after the end of the inhalative exposure. PMID- 8406926 TI - Blood lead and cadmium in a general population in Jinan City, China. AB - In 1991, blood samples were obtained from 150 adult Jinan citizens (74 men and 76 women at the ages of 20 to 57 years) who had no known occupational exposure to heavy metals. Age, sex, two social habits of smoking and drinking (in terms of daily consumption) and negative occupational history were examined in a medical interview. The samples were analyzed for lead (Pb-B) and cadmium (Cd-B) with a flame atomic absorption spectrometer. The geometric mean (GM) Pb-B and Cd-B were 92.3 and 0.94 micrograms/l, respectively, among 39 nonsmoking men, whereas the counterpart values were 123.4 micrograms/l and 2.61 micrograms/l among 35 smoking men (mean consumption; > 15 cigarettes/day); the difference was significant both for Pb-B and Cd-B. Comparison between 39 male and 76 female nonsmokers showed that Pb-B was significantly higher in men (92.3 micrograms/l) than in women (71.6 micrograms/l, whereas the difference in Cd-B (0.94 micrograms/l) for men versus 0.83 micrograms/l for women) was insignificant. When the women were classified by decade of age and Cd-B were compared, there was a trend of age-dependent increase in Cd-B from 0.60 micrograms/l in 20s to 1.24 micrograms/l in 40s, followed by no further increase at higher ages. Age-dependent changes were not remarkable in Pb B in women, or Cd-B and Pb-B in men. No significant time-dependent changes were observed when the present results were compared with the results from two similar studies conducted in 1983 and 1985, respectively. PMID- 8406927 TI - Reduction in cadmium in blood and dietary intake among general populations in Japan. AB - Blood samples and 24-h duplicates of food were collected in the years around 1980 and then in 1990's from inhabitants of nonpolluted regions in Japan. The 1990 study is still in progress with 286 blood samples and 125 food duplicates already collected from 7 regions. A preliminary analysis is made in the present study with the data obtained from the 7 study regions where the survey was conducted in 1980 and then in 1990, to examine possible changes in dietary cadmium intake (Cd D) and consequently cadmium levels in blood (Cd-B) during this 10 year period. The food duplicate analysis showed that Cd-D in 1980 was 47.1 micrograms/day (1.74):65 [GM (GSD):N] for men and 27.7 micrograms/day (1.75):65 for women. The counterpart values in 1990 were 29.5 micrograms/day (1.66):40 for men and 23.8 micrograms/day (1.73):85 for women; the difference between the two values for the same sex is significant (P < 0.05) both in men and in women, suggesting that there occurred about 37% and 14% reduction in dietary cadmium intake during this 10 year period in men and in women, respectively. Comparison in Cd-B showed that Cd-B for men and women in 1990 were 1.79 micrograms/l (2.01):121 and 1.84 micrograms/l (1.67):165, respectively, whereas the counterpart values in 1980 were 3.84 micrograms/l (1.64):123 and 3.57 micrograms/l (1.42):141, respectively, indicating significant (P < 0.01) reduction in Cd-B in both sexes in parallel to the reduction in Cd-D.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406928 TI - Biological markers as indicators of exposure and pneumoconiotic risk: prospective study. AB - This research is designed to evaluate a number of biological markers to estimate harmful exposure on coal miners from different mining regions in France and to relate the outcome to differences in prevalence of coal worker pneumoconiosis (CWP) between these regions. Eight epidemiological groups of active and ex-miners (smokers and non-smokers) have been selected in the French collieries (North, Lorraine and Provence) according to their occupational and pneumoconiotic status. The following biomarkers have been evaluated: cellularity of sputum, elementary analysis of particles in TEM/EDAX, plasma neutral metalloendo-peptidase elastase type (NMEP), leucocyte elastase (HLE), fibronectin (FN) and elastin peptides. Pulmonary alveolitis, expressed by sputum cellularity, is different between active workers groups but not related to the general background of pneumoconiosis prevalence in the French collieries. In the plasma parameters, fibronectin, HLE and NMEP significantly increased in all groups of coal mine workers as compared to the control group, except for fibronectin parameter in Lorraine collierie. The degree of increase of these parameters allow us to discriminate the different groups and suggest that plasma FN, HLE and NMEP may be considered as biological markers of chronic inhalation of coal mine dust particles. The decrease of elastin peptides level in the Lorraine group alone suggests a specific alteration of elastin metabolism. These parameters were not related to the development of pneumoconiosis and its degree of severity. PMID- 8406929 TI - Quantitative analysis of 8-hydroxyguanine in peripheral blood cells: an application for asbestosis patients. AB - The quantitative analysis of 8-hydroxy-2'-deoxyguanosine (oh8dG) in human peripheral blood cells was carried out to find integrated biomarkers for estimating cancer risk. The change of the oh8dG levels over time in two healthy volunteers was measured to evaluate a intraindividual variance and each individual value was confirmed to be almost constant when they maintained usual life style. We applied this measurement to asbestosis patients who had worked in dockyard for 19-42 years. The oh8dG were detected in all samples and ranged from 0.77 to 1.28/10(5) deoxyguanosine (dG)8. No significant differences was observed in mean values of oh8dG between patients (1.00 +/- 0.17/10(5)dG) and hospital control group without asbestos exposure (1.03 +/- 0.20/10(5)dG) No association was found with the status of cigarette smoking. The oh8dG level in peripheral blood cells is therefore not a sensitive biomarker for past asbestos exposure at low levels. PMID- 8406930 TI - Biological exposure indices of organic solvents for Korean workers. AB - Biological exposure indices (BEIs) of toluene, perchloroethylene (PCE) and methyl ethyl ketone (MEK) for Korean workers were studied respectively. Exposure in workplace to organic solvents were measured by personal sampling. Blood toluene, blood PCE, urinary trichloroacetic acid and urinary MEK were determined by headspace GC. Urinary hippuric acid was determined by HPLC and corrected for creatinine. BEIs for Korean workers were calculated with the levels of the determinants corresponding to permissible exposure limits in Korea which were the same with TLV of ACGIH. Blood toluene level of 2.2 mg/l and urinary hippuric acid level of 1.7 g/g creatinine corresponded to exposure of 100 ppm toluene. Blood PCE concentration of 1.6 mg/l and urinary trichloroacetic acid concentration of 2.9 mg/l corresponded to exposure to 50 ppm PCE. Urinary MEK concentration of 1.4 mg/l corresponded to exposure to 200 ppm of MEK. In conclusion, BEIs for Korean workers determined in this study were different to ACGIH's BEI as urinary determinants were lower and blood determinants were higher than ACGIH's BEI. PMID- 8406931 TI - In vivo differences between Asian, black and white in the stratum corneum barrier function. AB - The stratum corneum barrier function of Blacks, Caucasians and Asians were compared in vivo. A noninvasive technic, laser doppler velocimetry (LDV), was used to evaluate the cutaneous penetration of nicotinates by the determination of the lag time before vasodilatation induced by the application of those local vasodilator drugs. The study was performed on untreated skin and after removal of the stratum corneum by 12 strips. The influence of molecular weight and solubility of different nicotinates (methyl, ethyl, hexyl and vitamin E) were also studied on Japanese skin. Vasodilatation lag times assessed by LDV, with methyl nicotinate (MN), showed that skin permeability was more important in Asians (P < 0.01) and in Caucasians (P < 0.05) than in Blacks. Moreover Asian skin was significantly more sensitive to stripping (P < 0.05) than Black skin. A significant shorter lag time was obtained with small and hydrophilic nicotinates (methyl and ethyl) (P < 0.01) compared to a lipophilic one (hexyl). The alteration of the stratum corneum barrier function by stripping showed a more important modification with MN (P < 0.05) than with hexyl nicotinate. Consequently, this noninvasive method can evaluate the modifications of the stratum corneum barrier function and racial origin has to be taken into account in the determination of skin absorption. PMID- 8406932 TI - Modification of metabolism and neurotoxicity of hexane by co-exposure of toluene. AB - The effects of co-exposure of hexane and toluene were investigated in field surveys and animal experiments. One field survey suggested that increase of hexane content in adhesives might have caused an outbreak of polyneuropathy in a vinyl sandal manufacture in Japan. The animal experiments proved that co-exposure of hexane and toluene decrease hexane neurotoxicity and urinary excretion of hexane metabolites in rats. The results also suggested that toluene might inhibit metabolism of hexane. Another recent field survey indicated that the ratio of urinary 2,5-hexanedione to hexane exposure in the workers co-exposed to hexane and toluene decreased in parallel with in more crease of toluene concentration. The results indicated that urinary excretion of 2,5-hexanedione could be depressed by co-exposure of toluene even in the workers exposed to relatively low concentrations. These above-mentioned results suggest that co-exposure of hexane and toluene could inhibit hexane metabolism and decrease hexane neurotoxicity in both experimental animals and workers. Although metabolism of hexane could be easily modified by toluene or other solvents and might not be a good indicator for hexane exposure in mixed exposure, urinary 2,5-hexanedione might be a good indicator for neurotoxicity of hexane even in mixed exposure. PMID- 8406933 TI - Reference values for the study of low doses of metals. AB - The role of biological monitoring (BM) in the prevention of exposure to toxic metals has currently acquired considerable importance. However, in the near future the usefulness of BM in occupational medicine could be reduced by the fact that exposure to toxic substances will progressively decrease and consequently problems of comprehension will occur. BM can maintain its importance in the prevention field if sensitivity and specificity of the biological indicators (BI) are increased and if this activity is specifically directed towards groups of subjects. To achieve this last goal, one of the problems which must be solved is the identification of "reference values" (RV) which define the BI levels in the general population not occupationally or environmentally exposed to the toxic substance under study. It is necessary to "refer" to these values in order to compare the data obtained in a population which is presumed to be exposed. In this presentation, the example of blood cadmium is used in order to discuss the criteria required to identify RV. Herein the data of a personal case list are reported, particularly the variables which must be considered when selecting subjects to form "reference groups". Conversely, the possibility of obtaining RV through meta-analysis studies, pooling the results published in international literature, are evaluated. PMID- 8406934 TI - Quality assurance in the biological monitoring of lead exposure in China. AB - Based on the statistical data of the nationwide occupational diseases reporting in 1986-1988, lead poisoning stood the first place of chronic occupational poisonings in China. Biological monitoring of lead exposure is of paramount importance in both occupational and environmental health. In order to ensure lead testing data valid enough for taking actions of prevention and control, a quality assurance scheme including selection of reliable analytical techniques, establishing reference laboratory, application of standard reference materials, and conducting internal and external quality control program was implemented in China. Since the rigid assurance procedure was conducted during studies on "background levels of lead in non-exposure population" and "revision of the diagnostic criteria for lead", compatible and reliable results were obtained. PMID- 8406935 TI - Biological considerations in assessing exposures to genotoxic and carcinogenic agents. AB - This analysis of the relationship between long-term exposure to carcinogens and the risk of cancer relies upon a model which depicts exposure, burden and genetic damage as time series. It is shown that the dose of the carcinogen and the resulting damage to susceptible tissues can be related to cumulative exposure provided that linear kinetics are maintained and the exposure series remains stationary. Since saturable processes can lead to nonlinear behavior, the role of metabolism is investigated. It is argued that by maintaining the mean burden below 1/8 of the value of KM the contribution of nonlinear kinetics to the dose should be minimal, at least in occupational settings where saturable metabolism is most likely. Under this conjecture an expression is derived for the maximum mean air concentration, designated Xmax, which should maintain linear kinetics. By comparing values of Xmax, estimated for five genotoxic and/or carcinogenic substances (benzene, styrene, tetrachloroethylene, trichloroethylene, and vinyl chloride) with the corresponding U.S. occupational exposure limits, it is shown that saturable metabolism is unlikely to occur in some situations but is likely in others. This suggests that biological monitoring can play an important role in defining dose-response relationships for some, but not all, carcinogenic substances. PMID- 8406936 TI - Chromosome alterations in peripheral lymphocytes as indices of lifestyle and genotoxicity. AB - Short-term cultures of human lymphocytes were used to investigate the in vitro metabolism of benzene and its genotoxicity, and to monitor genetic health effects of lifestyles. Metabolic (S9) activation of benzene and its metabolites, catechol, hydroquinone, and phenol, caused an increase in sister-chromatid exchanges (SCEs) with different optimal concentrations of S9 mix for converting each compound into further reactive forms. The data indicate that catechol and hydroquinone can be optimally metabolized to produce reactive species, presumably benzo(semi)quinones, under conditions of lower metabolic activity than those necessary for phenol and benzene. We have further investigated the correlations between chromosome alterations (SCEs, structural aberrations and micronuclei) in peripheral lymphocytes and individual lifestyles. Healthy lifestyles, or "good health practices" examined were 1) not smoking, 2) not drinking too much alcohol, 3) doing physical exercise regularly, 4) sleeping more than 6 h per night, 5) keeping nutritional balance in meals, 6) not snacking, 7) having breakfast everyday, and 8) not having too much perceived stress. The persons were categorized into 3 groups having good, moderate and poor lifestyles by the number of good health practices they do. Mean frequencies of chromosome alterations in lymphocytes from men with poor lifestyles have been shown to be significantly higher than those in cells from men having good lifestyles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406937 TI - Biological monitoring of cadmium exposure in itai-itai disease epidemiology. AB - Cadmium (Cd) concentrations in blood and urine were significantly higher in itai itai disease patients, suspected patients and inhabitants in Cd-polluted areas in Japan. Urinary Cd concentrations were mainly related to the body burden of Cd and did not decrease for several years following cessation of Cd exposure. There was a close association between urinary Cd excretion and the occurrence of beta 2 microglobulinuria (beta 2-mg-uria). Probit regression analysis between urinary Cd excretion and, beta 2-mg-uria indicated that the threshold value may be set around 4 micrograms/g creatinine (cr.). Studies on inhabitants in Cd-polluted areas showed that urinary, beta 2-mg levels of more than 1000 micrograms/l or 1000 micrograms/g cr. were irreversible, while increased urinary excretion of beta 2-mg was associated with increased mortality. At present, beta 2-mg in urine is not a suitable indicator for monitoring renal tubular dysfunction in the general population since the significance of slightly increased excretion of urinary beta 2-mg has yet to be clarified. PMID- 8406938 TI - Applications of biological monitoring in occupational health practice: practical application of urinary 2-ethoxyacetic acid to assess exposure to 2-ethoxyethyl acetate in large format silk-screening operations. AB - A practical application of urinary 2-ethoxyacetic acid (EAA) to assess occupational exposure to 2-ethoxyethyl acetate (EGEE-Ac) during a large format silk-screening operation is described. Industrial hygiene air monitoring of employees of a silk-screen shop producing large aircraft interior panel coverings revealed a broad range of exposures to EGEE-Ac. Time weighted exposures averaged 12 ppm (range 2.9-34 ppm) in press operators during production press runs, exceeding the 5 ppm Washington State permissible exposure limit. Employees were instructed to use organic vapor respirators until engineering controls could be developed. Urinary monitoring of EAA was conducted on 30 employees by the company medical department to aid in exposure risk assessment and to assess compliance. Results obtained ranged from 1.1-27 mg EAA/g creatinine which compares favorably with the proposed Biological Exposure Index (BEI) of 100 mg EAA/g creatinine. Results of representative air and biological monitoring, and observations of work practices for different exposure groups indicated that inhalation exposure was the predominant route of exposure. Follow-up testing to assess the efficacy of a newly installed ventilation upgrade is planned. PMID- 8406939 TI - Pharmacokinetic modeling as a tool for biological monitoring. AB - The relationships between biological indicators and exposure or tissue burdens are determined by the pharmacokinetic behaviour of the chemical. They can be studied by pharmacokinetic models of various types. Simple pharmacokinetic models are used here to describe general relationships valid for large groups of chemicals or situations. Important parameters to consider are the half-life of the biological indicator, the individual variability and the exposure variability. Biological sampling strategies are presented for monitoring of groups of workers, or individual workers. For specific chemicals, mainly solvents, more elaborate models can be developed, i.e., physiologically-based pharmacokinetic models including physiological, metabolic and physicochemical parameters. Such models are useful to describe the influence of confounding factors. Physiologically-based pharmacokinetic models can also be developed for metals and metalloids. Antimony is presented here as an example. In conclusion, pharmacokinetic modeling brings much information on sampling time, sample size, limit values, effect of physical workload and of individual physiological parameters. PMID- 8406940 TI - Confounding factors in biological monitoring of exposure to organic solvents. AB - Some environmental and physiological factors that affect the toxicokinetic behaviors of organic solvents were examined using a physiologically based pharmacokinetic model developed for trichloroethylene (TRI). The conclusions are as follows: 1) Body fat content substantially affects the kinetic behavior of TRI: the TRI concentration in blood and the urinary excretion rate of its metabolites are higher in slim men than in obese men during exposure, but these parameters eventually become higher in obese men. 2) There is a sex difference in the pharmacokinetic profiles of TRI. Although retention of TRI in the body is greater in men than in women, the blood concentration of TRI in women is higher than in men 16 hours after exposure. 3) Because of increased pulmonary ventilation and cardiac output, physical activity (workload) during exposure greatly increases the blood concentration of TRI and the urinary excretion of its metabolites, whereas the activity after exposure has only a marginal influence. 4) Ethanol-induced inhibition of TRI metabolism causes a marked change in the kinetic behavior when the TRI exposure level is low, whereas enzyme induction by ethanol significantly affects the kinetics only when the exposure concentration is high. The effect of enzyme induction differs from compound to compound. Whether the compound concerned is a perfusion-limited substrate (TRI, for example) or a capacity-limited substrate (1,1,1-trichloroethane, for example) determines the extent of the effect. 5) Metabolic interaction between solvent vapors inhaled simultaneously may not become apparent until the exposure level increases to a degree that will overload the enzyme capacity. PMID- 8406941 TI - Biological monitoring of occupational pesticides exposure. AB - Two kinds of measurement: (1) enzyme activities in blood, and (2) unchanged pesticides and their metabolites in urine or blood have been used in biological monitoring for assessing exposure to pesticides. The assays of acetylcholinesterase (AChE) activity in whole blood and erythrocytes are mainly applied to estimate inhibition by organophosphates (OPs) and carbamates. A level at 70% of an individual's baseline or of a mean population AChE activity has been recommended as a reference value for exposure control. The measurement of lymphocyte "neuropathy target esterase (NTE)" activity in subjects handling axonopathic OPs is mainly for research application. Analytical methods are available for detecting alkylphosphates, carbamates, pyrethroids, chlorinated hydrocarbons, some herbicides and fungicides, chlordimeform, chlorobenzilate, dichloropropene, dinitrocresol and pentochlorophenol or their metabolites in urine or blood. However, due to lack of significant dose-response or dose-effect relationship, the majority of these determinants can only be used as biological exposure indicators to confirm exposure or to estimate internal dose. Further research in developing adequate indicators and methods for biological monitoring of occupational pesticides exposure is needed. Pre-exposure value and/or reference value of relevant indicators are necessary for assessing the degree of exposure and absorption. PMID- 8406942 TI - Use of biologic markers for toxic end-points in assessment of risks from exposure to chemicals. AB - Biologic markers are familiar tools for monitoring human absorption of, and reaction to, potentially toxic chemicals. The concept of applying biologic markers to the risk assessment process is a natural, but more recent, development and the principles remain to be fully elaborated. Biologic markers may be measurements of exposure, of effects, of genetic or induced sensitivity or of disease. The ideal biologic marker for risk assessment purposes is a quantitative measurement of a chemical, biochemical, functional or morphological change in the system that is initiated by a chemical and which results in pathologic change and overt toxicity. It follows that some understanding of the mechanism of toxicity and of dose-response relationships are pre-requisite for selection of suitable biologic markers for use in risk assessment. Where biologic markers for toxicity are common between mammalian species, extrapolation of data for quantitative risk assessment purposes becomes more reasonable. In the field of carcinogenesis, some DNA and protein adducts have been proposed as biologic markers for assessment of risk associated with exposure to genotoxic carcinogens. However, less progress is evident in relation to other toxic end-points including those for pulmonary, reproductive, immuno- and neuro-toxicity, despite intensive efforts. PMID- 8406943 TI - Quality assurance of biological monitoring in view of risk management. AB - Since recommended occupational exposure limits both for workplace air concentration and biological monitoring results generally have very narrow margin of protection, the probability of the false decision due to measurement error should be perceived and carefully controlled. Japan Federation of Occupational Health Organization has conducted large scale quality control programs since 1987 and reported the results in detail. The meta-analysis of the report of 1992 made it clear that the qualities of the analysis of the leading laboratories as expressed by the coefficient of variation of measurement error are mostly about 5% or less. It was suggested that the warning level should be set at (biological exposure limits minus 2 SD of the measurement error). However, it will not be needed when the SD is less than 2.5% as was found for the analyses of blood lead, urinary hippuric acid, methyl hippuric acid, and mandelic acid by a few best performing laboratories. PMID- 8406944 TI - Metals in hair as biological indices for exposure. AB - Hair samples from three groups of occupationally exposed subjects were analyzed for their lead (Pb), manganese (Mn) and mercury (Hg) contents. For lead (number of subjects, n = 209), the hair Pb ranged from 0.93 to 3527 micrograms/g (geometric mean, GM = 641) and blood Pb from 33.3 to 774 micrograms/l (GM = 341); for manganese (n = 38), the hair Mn ranged from 0.20 to 52.97 micrograms/g (GM = 2.66) and urine Mn ranged from 1.70 to 17.9 micrograms/l (GM = 5.56); and for mercury (n = 85), the hair Hg from 1.79 to 12.8 micrograms/g (GM = 5.09) and the blood Hg from 0.63 to 57.3 micrograms/l (GM = 10.9). The hair Pb was significantly (P < 0.0001) correlated to blood Pb (r = 0.85); the hair Mn to urinary Mn (r = 0.45); and the hair Hg to blood Hg (r = 0.53). The average metal content at the distal end was not significantly (P > 0.05) different from that of proximal end. The GM levels for the distal end were 223 micrograms/g (95% CI 152 347) and 2.26 (95% CI 0.97-5.29); and those for the proximal end were 186 (95% CI 97-261) and 1.18 (95% CI 0.54-2.58) for Pb and Mn respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406945 TI - Development and utilization of a new simple assay for DNA-protein crosslinks as a biomarker of exposure to welding fumes. AB - A new method for DNA-protein crosslinks involving selective precipitation of DNA containing crosslinked proteins by K+ sodium dodecyl sulfate was utilized in the peripheral white blood cells of 21 male metal arc welders and in 26 male controls of similar age and racial characteristics who were not exposed to welding fumes. DNA was quantitated by Hoescht fluorescence. Although the concentration of nickel and chromium in the peripheral blood was low and did not differ between subjects in the two groups, one-fourth of the welders had levels of DNA-protein crosslinks that were above the upper limit of the controls. Mean crosslink values were 1.85% (+/- 1.14) among the welders and 1.17% (+/- 0.46) among the controls, a 58%, statistically significant difference (P = 0.01). Thus, many welders appeared to be burdened with an excess of DNA-protein crosslinks suggesting exposure to crosslinking agents and, possibly, a detectable biologic effect of potential genotoxic consequences. PMID- 8406946 TI - The correlation between blood and urine level of cadmium and nasal and paranasal sinuses disorders. AB - Over exposure to cadmium may produce numerous adverse health effects, including changes in the nasal mucosa. Cadmium levels were measured in blood and urine samples taken from 106 workers who were exposed to high levels of cadmium and nickel in a cadmium nickel battery plant. Pathological results were observed in 46% and 51% of the blood and urine samples respectively. A positive association was found between clinical nasal complaints and pathological levels of cadmium in blood (P = 0.006) and urine (P = 0.045) samples. All the workers underwent sinus X-rays and 63% of them were abnormal. No correlation was found between pathological sinus X-rays and pathological cadmium level in blood and urine samples. A positive correlation was found between clinical nasal complaints and abnormal sinus X-rays. The prevalence of sinus X-ray abnormalities in an asymptomatic general population is 33% as compared to 63% in our study. This difference is highly significant statistically (P < 0.0001). Our result shows a possible harmful effect of cadmium on the paranasal sinuses. PMID- 8406947 TI - Urine, serum and hair monitoring of hydrofluoric acid workers. AB - To define the relationship between fluoride (F) concentration in the serum, urine and hair of workers and the concentration of hydrofluoric acid (HF) in the work environment, pre- and postshift serum and urine samples of 142 HF exposed workers and 237 unexposed workers were examined. Hair specimens were also collected for the determination of F. To determine whether external contamination influences hair analysis, the control hair samples were kept in the work environment for one week. The pre-exposure levels in serum and urinary F in HF workers were higher (P < 0.01) than the control values. This suggests that F excretion from the body continues for at least 12 hours. The postshift serum and urinary F concentrations of these workers were significantly higher (P < 0.01) than the preshift concentrations. The levels of F in the hair of HF workers were also higher than in the control subjects. The concentrations of F in postshift serum and urine, and hair were in good correlation to each other. There was a linear relationship between mean serum and urinary F concentrations and HF concentration in the workplace. A mean F concentration of 82.3 micrograms/l in serum and 4 mg/l in urine with a lower fiducial limit (95%, P = 0.05) of 57.9 micrograms/l in serum and 2 mg in urine were estimated to correspond to an atmospheric HF concentration of 3 ppm, which is the maximum allowable concentration recommended by Japan Association of Industrial Health and also the threshold limit value suggested by American Conference of Governmental Industrial Hygienists.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406948 TI - 32P-postlabeling analysis of dibenz[a,j]acridine DNA adducts in mice: preliminary determination of initial genotoxic metabolites and their effect on biomarker levels. AB - N-Heterocyclic aromatics (NHA) are widely occurring environmental pollutants formed during the pyrolysis of nitrogen-containing organic chemicals. NHA are found in significant amounts in tobacco condensates, synthetic fuels, gasoline engine exhaust, and effluents from the heating of coal. Dibenz[a,j]acridine (DBA) is an example of NHA. The potency of many carcinogenic compounds is related, at least in part, to the efficiency of their biological activation. We undertook studies to determine which initial metabolites of DBA lead to the formation of high levels of carcinogen-DNA adducts in vivo. DBA and its metabolites, trans-DBA 1,2-dihydrodiol (DBA-1,2-DHD), trans-DBA-3,4-dihydrodiol (DBA-3,4-DHD), and trans DBA-5,6-dihydrodiol (DBA-5,6-DHD), were applied to the skin of mice. DNA was isolated using enzyme-solvent extraction method. DNA was 32P-postlabeled under conditions of limiting [32P]ATP. In skin, DBA produced two distinct adducts. The same two adducts were seen when DBA-3,4-DHD was applied. In addition the total adduct level elicited by DBA-3,4-DHD was higher than that of parent compound. Two adducts were seen when DBA-5,6DHD was applied, but these were very different from adducts seen with DBA. These results suggested that activation of DBA to DNA binding compounds in skin includes initial formation of DBA-3,4-DHD. The data support development of biomarkers for the exposure and effect of this compound, and also suggest that specific metabolic susceptibility markers might be able to predict populations at increased risk. PMID- 8406949 TI - The dental health of adults in an integrated urban development in Addis Ababa, Ethiopia. AB - Two hundred and forty-three Ethiopian adults (18 years-old and over) were examined for caries, periodontal disease, malocclusions and enamel opacities. These adults were the parents of children cared for by an independent charitable organisation, the Ethiopian Gemini Trust. The prevalence of dental caries was generally low with a mean DMFT for the sample of 2.7 (+/- 0.2) and a mean DMFS of 6.7 (+/- 0.6), although one adult had a DMFS of 62. A high proportion of the adults (83.5 per cent) had calculus, but only 2 per cent had deep pocketing. Twenty-three per cent of the adults had a malocclusion and for 6 per cent of these this was moderate to severe. The most prevalent enamel defects were hypoplasias and diffuse opacities with 22 per cent of adults having one or more index teeth affected. Access to dental services was virtually non-existent as judged by the clinical status of these adults. PMID- 8406950 TI - Various reasons for permanent tooth extractions in a Caribbean population- Antigua. AB - A study was undertaken in September 1990 to investigate the reasons for permanent tooth extractions in Antigua. Eight out of 10 dentists on the island were requested to record permanent tooth extractions, including tooth type and reasons for extraction, during a 6 week study period. All 8 dentists provided data which indicated that 471 teeth were Extractions for orthodontic and impaction reasons comprised 4.3 per cent of tooth extrac-irregularly attending dental services. The principle reason for extractions was caries (61.6 per cent). Periodontitis (29.9 per cent) was the next most frequent reason for extractions and it became the predominant indication for tooth loss after 40 years of age. Extractions for orthodontic and impaction reasons comprised 4.3 per cent of tooth extraction, but these causes disappeared after 30 years of age. Restorable teeth were not commonly removed. The tooth types most frequently extracted due to caries were lower first molars and upper molars. Most commonly extracted, periodontally involved teeth were lower central incisors and upper third molars. PMID- 8406951 TI - Systemic diseases in elderly dental patients. AB - Systemic diseases, blood pressure and pulse rate, were investigated in 1012 elderly patients who consulted the gerodontic clinic of Tokyo Medical and Dental University Hospital. One or more systemic diseases was found in 64.2 per cent of the subjects. Hypertension was the most frequent systemic disease, occurring in 30.9 per cent of the subjects, and was followed by angina pectoris, diabetes, arrhythmias, digestive diseases, cerebrovascular diseases and myocardial infarction, respectively. Nearly 17 per cent of the patients had two or more diseases. Among these diseases, hypertension was frequently combined with angina pectoris, myocardial infarction, arrhythmia, and cerebrovascular diseases. Patients with systemic diseases are at a greater risk during dental treatment. Psychosomatic stress and vasoconstrictors in local anaesthesia possibly induce the exacerbation of systemic diseases or unexpected cardiovascular reactions. Isolated systolic hypertension (systolic blood pressure of 160 mm Hg or more) was noted in 14.3 per cent of the patients. Among these patients, 6.3 per cent of patients were hypertensive patients who had both above 160 mm Hg systolic pressure and 95 mm Hg diastolic pressure. Elderly dental patients have various systemic diseases. Accordingly, it is very important for the dentist to identify systemic diseases of patients and to understand systemic conditions before treatment commences. PMID- 8406952 TI - The Cambodian National Oral Health Plan 1992-2000. AB - The reconstruction of dental services and training in Cambodia following the devastation of the country during the reign of the Khmer Rouge has been limited by minimal help from other countries for political reasons. Following the recent signing of a peace treaty, a National Conference on Oral Health has been held, and a National Oral Health Plan formulated, outlining goals for the year 2000, and strategies to improve the oral health status in the country. PMID- 8406953 TI - New ideas and advancing technology in prevention and non-surgical treatment of periodontal disease. AB - Analytical, oral epidemiological studies in adult 'toothbrushing' populations show that the highest prevalence of missing teeth is for molars and maxillary premolars ('key-risk' teeth). The highest prevalence of subgingival microflora, gingivitis (CPITN-1), plaque retentive factors, such as subgingival calculus and restoration overhangs (CPITN-2), and diseased pockets > 3 mm (CPITN-3-4) is found on the approximal surfaces of the same teeth ('key-risk' surfaces). Primary and secondary prevention, as well as treatment of periodontal disease should be focused on these 'key-risk' surfaces. One single, well-executed subgingival scaling and root-planning procedure in deep, diseased periodontal pockets, followed by oral hygiene training and professional mechanical tooth-cleaning (PMTC) at need-related intervals, will prevent further loss of periodontal attachment. However, if the root cementum is removed during instrumentation, and the post-treatment plaque control programme fails, microorganisms will recolonize and invade the rough exposed root dentine, resulting in recurrence of periodontitis and possibly, root caries and pulpitis. Some of these problems may be overcome by the application of new instruments and methods for self-care, PMTC, removal of overhangs, scaling and root-planing without removing 'non diseased' cementum. PMID- 8406954 TI - Paediatric dentistry: Fauchard and before. AB - Although conventional wisdom suggests that paediatric dentistry was not practised until the early twentieth century it was in fact recorded in much earlier times. The authors detail many authors and books from previous centuries concerned with weaning, infant feeding, remedies for teething pains and eruption. Further details include the early advice on the avoidance of caries and the beginnings of the specialty of orthodontics. PMID- 8406955 TI - Titanium and titanium alloys as dental materials. AB - Because of light weight, high strength to weight ratio, low modulus of elasticity, and excellent corrosion resistance, titanium and some of its alloys have been important materials for the aerospace industry since the 1950s. Now, with the additional advantages of excellent biocompatibility, good local spot weldability, and easy shaping and finishing by a number of mechanical and electrochemical processes, these materials are finding uses in dental applications, such as implants and restorative castings. Although more research is still needed in areas such as development of optimal casting investments, porcelain veneering systems, device designs, and controlled biological responses, the present and future uses of titanium appear bright for dentistry. PMID- 8406956 TI - On statistics of success and loss for dental implants. AB - Assessments of implantation success have mainly been based on incorrect statistical methods: the binomial approach of percentage of implants not failed to-date over inserted implants. The survival or life-table approach, instead, adjusts individually for duration of exposure (from insertion until explantation or last recall examination). If published failure percentages are adjusted for mean time of follow-up, which is reported in at least some papers, and a lower 95 per cent-confidence bound on the failure rate is calculated, considerably lower probabilities of one, two or more years of survival will be obtained in many cases. An analysis of 2,292 implants yielded a pre-prosthetic 6-months survival probability between 87 and 91 per cent. The post-prosthetic 2-years survival for pillar-shaped (n = 314 with single tooth replacements) and screw implants (n = 150 with edentulous jaws and one implant, randomly chosen, per patient) may be between 79 and 89 per cent and 79 to 96 per cent, respectively (95 per cent confidence intervals). In pillar-shaped implants, there is preliminary evidence that ceramic implants and male patients carry a larger risk of failure than metal implants and female patients. With screw implants, only high patient age at implantation may be a risk factor. PMID- 8406957 TI - Review of behavioural research in dentistry 1987-1992: dental anxiety, dentist patient relationship, compliance and dental attendance. AB - A review is presented of the behavioural research in dentistry published in or after 1987 on dental anxiety, dentist-patient relationship, compliance and dental attendance. It is concluded that little progress has been made in the field of behavioural dentistry during those years and recommendations are made to improve future research efforts. PMID- 8406958 TI - Social and psychological theories and their use for dental practice. AB - This paper discusses four major theoretical constructs that have helped broaden our understanding of oral hygiene behaviours and dental health risks, including: social class, life stress, self-efficacy and locus of control. The potential of these constructs to enhance the treatment of dental diseases is illustrated using data from a study based on a biopsychosocial model of caries risk. In two cities in Connecticut, 481 children aged 3 years were recruited from the Head Start programme. Two dentists examined the children with a mirror and explorer and collected data on dmfs. A parent or guardian of 369 of these children completed an interview with a research assistant which provided data on each child's brushing habits, dietary intake of sugar, social class indicators, stressful life events, dental health locus of control and perceived dental self efficacy. The effects of psychosocial variables on two clinical measures were assessed, the levels of S. mutans and the prevalence of decay. Analysis indicated that S. mutans was the most important predictor of caries risk. Mothers who had more external locus of control beliefs, lower income, more knowledge about tooth decay and higher stress levels had children with greater risk of having caries. Further, use of a baby bottle at night was a significant factor in caries risk. Analyses of S. mutans risk showed that socioeconomic status and ethnicity significantly differentiated those with no colonies from those with one or more colonies while attitudinal and behaviour factors predicted who would be in the group with the highest number of S. mutans colonies. PMID- 8406959 TI - Dental fears in general practice: new guidelines for assessment and treatment. AB - Dental fear is a world-wide problem and universal barrier to oral health care services. Fears acquired in childhood through direct experience with painful treatment or vicariously through parents, friends or siblings, may persist into adulthood. Symptomatic treatment and lack of trust and control exacerbate fears. Coexisting psychiatric conditions further complicate treatment. Treatment steps recommended in this paper include careful assessment of fears, application of dental and psychological management techniques to enhance trust, feelings of control, and the development of coping skills, and appropriate and targeted use of pharmacological agents. Prevention of fear development through the use of effective behavioural child management techniques combined with preventive dentistry should be a fundamental part of general dental practice throughout the world. PMID- 8406960 TI - Factors affecting the compliance of patients with preventive dental regimens. AB - The common dental problems of caries and periodontal disease can be controlled by patients who are prepared to adopt the appropriate dietary and oral hygiene behaviours. However compliance with dental surgery based health education programmes is often disappointing. The failure of these interventions is due to the fact that the majority of the dental profession are naive and have a rather insular approach to patient education. There are two key influences on the patient. First there is the 'macro environment' whereby behaviour is shaped by the norms on lifestyle which exist in a patient's social environment. Behaviour changes which are a variance with these norms will not be successful, therefore health education programmes must take cognisance of these norms. It may be necessary to change certain oral health behaviours slowly over an extended period of time. Second there is the 'micro environment' in the dental office. All too often patient education is unplanned, haphazard, not relevant to a particular patient and difficult to understand. Dental professionals should give prevention the same status as clinical care so that it is well planned and carefully evaluated. PMID- 8406961 TI - Acceptance of full dentures. AB - The wearing of even technically perfect full dentures is associated with a significant deterioration of most, if not all, oral functions. The fact that between 85 and 90 per cent of patients receiving new and technically well made dentures and between 65 and 75 per cent of patients with older dentures are generally satisfied with the treatment result is therefore more of a tribute to human adaptation than to prosthodontic skill. Research has shown that variables claimed to play an important role in patient acceptance of full dentures, such as the quality of the dentures, the oral condition, the patient-dentist relationship, the patients' attitude toward dentures, the patients' personality, socio-economic factors, demographic variables, previous denture experience and oral stereognosis, have in fact no or negligible effects in this respect. Nor has the degree of prediction been appreciably improved by combining the effects of such variables. Variables of theoretical importance for acceptance of dentures such as the patients' motory ability and the quality and quantity of saliva have still to be studied. At the present time research has yet failed to furnish the clinician with a practical method whereby patients who are unlikely to accept full dentures can be identified with any degree of certainty. The clinician has therefore still no option but to base his prediction on an individual estimate of his patient's adaptive ability. PMID- 8406962 TI - Morphology of the root canal wall and arrangement of underlying dentinal tubules. AB - The morphology of the root canal wall and the arrangement of the dentinal tubules underlying it were investigated in this study. Fifty clinically sound single rooted human teeth were cut for longitudinal and transverse sections. After dissolving the organic components and removing the smear layer, the specimens were prepared for scanning electron microscopy. In the upper root level, semiglobular calcospherites were present on the canal wall and a convergent arrangement of the dentinal tubules was found on the transverse sections. At the mid-root level, an irregular area of calcification comprising of compressed calcospherites began to appear on the proximal wall and increased towards the apex. The transverse sections showed the crowding and convoluted arrangement of the dentinal tubules near the canal wall, in the proximal area. A correlation was noticed between the irregular calcification on the wall and a convoluted tubular arrangement below the wall. These features were common in most of the specimens. This irregular secondary dentine does not appear to be pathological but physiological: associated with the normal ageing of the root. PMID- 8406963 TI - Comparison of two sonic handpieces during the preparation of simulated root canals. AB - The objective of this study was to assess and compare two sonic handpieces during the preparation of simulated root canals. A total of 60 simulated canals in clear resin blocks of various angles and positions of curvature were prepared using Shaper files activated either by an MM1500 Sonic Air or an MM1400 Mecasonic handpiece. Each handpiece was used to prepare 30 canals using an identical preparation procedure which involved a linear filing motion and an anticurvature technique in curved canals. The efficacy of the handpieces was determined for straight and curved canals separately and included an assessment of loss of canal length, weight loss from the blocks, smoothness of canal walls, transportation and overall shape of the prepared canals. Canals were prepared rapidly and effectively by both handpieces. Overall, the MM1500 handpiece was associated with significantly less distance loss (P < 0.05) and significantly more loss of weight (P < 0.01). Only four prepared canals (8%) had hour-glass shapes displaying zips and elbows, each handpiece created two. Only two canals (4%) had danger zones, both were created by the MM1400 handpiece. There was little difference between the handpieces in terms of the smoothness of canal walls, the direction and amount of transportation or the overall shape of canals. Under the conditions of this study, the new MM1400 Mecasonic handpiece performed as well as the established MM1500 Sonic Air. PMID- 8406964 TI - Residue of gutta-percha and a glass ionomer cement sealer following root canal retreatment. AB - A previous report indicates that root canals obturated with a new glass ionomer cement endodontic sealer (Ketac-Endo, Espe, Germany) and laterally condensed gutta-percha can be retreated by ultrasonic instrumentation. To address the possibility of Ketac-Endo being used without condensation of gutta-percha, the purpose of this study was to evaluate the efficacy of ultrasonic retreatment in canals obturated with single-cone gutta-percha and Ketac-Endo. Thirty root canals were prepared in a standardized way to ISO size 40 and obturated with gutta percha and Ketac-Endo. Either a size 40 gutta-percha cone was used with and without lateral condensation, or a size 25 single cone, without condensation. After 14 days, the canals were retreated using chloroform and ultrasonic instrumentation. The roots were split vertically, and the amount of residual debris on the canal walls was assessed by three examiners using a dissecting microscope. Debris was recorded in the apical, middle and coronal canal levels according to a preset evaluation scale. The mean scores for each group were compared by ANOVA and Mann-Whitney U-test, with a 5% level of significance. In the roots obturated with lateral condensation the amounts of residual debris in the coronal and middle canal levels was lower than in the other two groups, whereas in the apical level it was higher. Statistically, only the differences in the apical level were significant (P < 0.03). It was concluded that ultrasonic retreatment may be performed effectively in root canals obturated with single cone gutta-percha and Ketac-Endo. PMID- 8406965 TI - A assessment of the plastic Thermafil obturation technique. Part 1. Radiographic evaluation of adaptation and placement. AB - Adaptation and placement of alpha-phase gutta-percha delivered with a plastic core-carrier, Thermafil, was compared to the lateral condensation of gutta-percha in a specific tooth model. Fifty-one mandibular molar roots with separate canals, patent canal orifices and curvatures greater than 15 degrees were cleaned and shaped with K-files and 2.5% sodium hypochlorite to a size 30 at the apex, and flared with Hedstrom files to create a continuously tapering funnel preparation. Canals were randomly obturated with Sealapex root canal sealer and either alpha phase gutta-percha on a plastic Thermafil carrier, or standard beta-phase gutta percha with lateral condensation. Roots were radiographed from the proximal and evaluated by three examiners, based on established criteria for overall material adaptation, apical adaptation, and filling material extrusion. Thermafil provided a statistically significant better overall canal obturation (P < 0.001), while, in the apical third, both techniques were not significantly different (P > 0.05). When the apical orifice was patent there was a significant propensity for the extrusion of filling materials beyond the apex (P < 0.001) with the Thermafil technique. PMID- 8406966 TI - An assessment of the plastic Thermafil obturation technique. Part 2. Material adaptation and sealability. AB - The short- and long-term apical seal of root canals obturated with softened alpha phase gutta-percha on plastic core-carriers, Thermafil, was compared to that of laterally condensed, cold gutta-percha. Fifty-one roots from mandibular molars with separate canals, patent canal orifices and curvatures greater than 15 degrees were cleaned and shaped with K-files and 2.5% sodium hypochlorite to a size 30 at the apex, and flared with Hedstrom files to create a continuously tapering funnel preparation. Canals were randomly obturated with Sealapex root canal sealer and either alpha-phase gutta-percha on a plastic Thermafil carrier, or standard beta-phase gutta-percha with lateral condensation. Teeth were separated into three groups of 17 each and placed in black India ink for 24 h, 7 days or after 5 months storage in water. The teeth were demineralized, rendered transparent, and apical microleakage determined by the linear measurement of dye penetration. Significant differences in microleakage were noted between the 24 h and 5-month Thermafil groups (P < 0.05), 24-h and 7-day lateral condensation groups (P < 0.05), and 24-h and 5-month lateral condensation groups (P < 0.05). There were no significant differences between the two techniques at each time interval. It was concluded that both techniques demonstrated a significant increase in apical microleakage over a 5-month period. PMID- 8406967 TI - An assessment of the plastic Thermafil obturation technique. Part 3. The effect of post space preparation on the apical seal. AB - The effect of post space preparation on the apical seal of teeth roots treated with the plastic-cored Thermafil was studied in vitro. Single rooted teeth with mature apices were prepared chemomechanically and obturated with alpha-phase gutta-percha, Thermafil, on plastic core-carriers and sealer, Sealapex. The teeth were divided randomly into four groups of 18 teeth each. In two of the groups post space was prepared by hand using a Parapost drill (diam. 1.25 mm) after gutta-percha removal with Peeso reamers in a slow-speed handpiece, one group immediately after canal obturation and the other after storage in 100% relative humidity at 37 degrees C for 1 week; 5 mm of root filling was left intact. The time taken to remove the gutta-percha was recorded. Post space was not prepared in the control groups. Apical leakage was determined using black India ink. The teeth were demineralized, rendered transparent, and linear measurement of dye penetration was recorded. Results showed that there were no significant differences in leakage between any of the groups (P > 0.05). There was no statistically significant difference in the time required to remove the gutta percha from the root canals in both groups (P < 0.05). It is concluded that under the conditions of this study post space preparation immediately after obturation, or after a delay of 1 week, does not affect the apical seal of Thermafil with plastic core-carriers. PMID- 8406968 TI - The influence of different engine-driven, sound ultrasound systems and the Canal Master on root canal preparation: an in vitro study. AB - Different methods and automated devices have been developed to accomplish the enlarging and debriding of root canals. The occurrence of undesired effects on the root canal walls was compared in vitro between a theoretically ideally shaped canal and the Canal Leader, Canal Finder, 3000 Endo Sonic Air, Cavi Endo 25, Enac System, Piezon Master 400 and the Canal Master. Clear casting resin models were used for this purpose. A photographic double exposure was used to superimpose the images of the original canals on the enlarged canals, thus enabling measurement of the material removed. Measurements were made at six different points. Significance was defined at the 5% level (P = 0.05). All instruments and/or devices, with the exception of the Cavi Endo 25, enlarged the convex side of the canals significantly greater at level 1 in comparison to the ideal. On the concave side, significantly smaller differences were found between the ideal and the results of all the enlarging devices, with the exception of the Canal Master. On the convex side at level 2, the Canal Leader, Canal Finder, Cavi Endo, Enac System and Endo Sonic Air showed no statistical differences between ideal and effective material removal. The Piezon Master showed significantly lower material removal in comparison to the ideal. The Canal Master demonstrated statistically higher differences when compared with the ideal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8406969 TI - Constructing success and failure: age differences in perceptions and explanations of success and failure. AB - Age differences in attributions for self-reported successes and failures in both important and daily situations were examined. Sixty-one young adults (M age = 19.2), twenty-one middle-aged adults (M age = 45), and fifteen older adults (M age = 71.4) gave attributions and affects for self-chosen situations, which were classified as social or nonsocial. The attributions and affects were coded according to Blank's scheme with attributions dichotomized into internal and external, and stable and unstable. Middle-age and older adults were more likely than young adults to attribute failure to external causes and to describe more social than nonsocial situations. Consistent age differences in attributional stability were not found nor were there age differences in attributional internality for success outcomes. PMID- 8406970 TI - Aging couples and disability management. AB - The purpose of this work is to identify, in an elderly population, couple patterns and to relate this typology to the management of disabilities. Resulting from questions arising from research in gerontology, it utilizes the theoretical contribution of the sociology of the family so as to understand the relationships between the functioning of the couple and the management of disability. Four types of couples have been identified according to the presence or absence of a tacit contract between husband and wife and according to the character of their relationships. In each type, we have studied the conduct of daily life and its negotiation as well as the management of disabilities. The analysis shows that this management is very different according to the type of couple and thus confirms the importance of taking the functioning of the couple into account so as to fully understand the situation of disabled married elderly persons. PMID- 8406971 TI - Open-ended attributions for the performance of the elderly. AB - The present study explored open-ended attributions for the success and failure of relatively younger and older men in social and academic situations using a between-subjects design. Attributions were collected from 109 college students and were coded using the Elig and Frieze scheme. Results showed that respondents were more likely to make attributions that combined age with other attributional categories (age-related attributions) than attributions solely to target's age. And, they made more ability-task interaction than ability attributions. Moreover, both attributions were as likely to be made for the elderly adult's success as his failure. Finally, differences also emerged in results from the stability dimension and those for age-related and ability-task attributions indicating that these parameters should be assessed independently in future research. PMID- 8406972 TI - Cross-generational attributions concerning locus of control beliefs. AB - Age differences and attributions of age differences in locus of control orientation were examined for college students and older adults. Self-ratings and ratings of the other group by sixty college students and ninety-seven older adults were measured using Rotter's locus of control items in a Likert format. Findings showed that both age groups misattributed levels of control orientation to the other group when compared to that group's self-ratings. Older adults endorsed more internal beliefs than did college students. College students viewed older adults as more external than older adults viewed themselves, while older adults viewed college students as more internal than students' self-ratings indicated. The common perception in gerontological literature that older adults are particularly external in their locus of control beliefs may represent an erroneous attribution rather than self-reported beliefs. PMID- 8406973 TI - "The play's the thing": Samuel Beckett's midlife transition and the Theatre of the absurd. AB - This article considers the question of shifts in artistic and literary style from a lifespan developmental perspective, focussing on changes during midlife. Using Samuel Beckett as an example, we demonstrate the relation between his abrupt shift to playwriting (and specifically to the Theatre of the Absurd) and his entrance into midlife. Abandoning the "counting the wrinkles" approach to mid adulthood, we consider midlife change in terms of growth. We base our contention on theories of lifespan personality development that suggest parallels between the technical demands of dramatic art and the psychological needs of midlife. PMID- 8406974 TI - Age and forgetfulness: young perceivers' impressions of young and older neighbors. AB - Previous person perception research has indicated that people use an age-based double standard when judging targets who experience single instances of memory failure. The two experiments reported here extend the investigation by assessing whether perceivers evidence a similar bias in judging both the memory capability and personal traits of targets who vary in age and degree of forgetfulness. In the first experiment 179 young adults rated how likely they would be to choose a certain type of neighbor, described in a two-page vignette, to perform memory tasks. The neighbor's (i.e., target person's) age and degree of forgetfulness were manipulated. Participants gave higher choice ratings to nonforgetful than to forgetful targets. Also, they gave higher choice ratings to old over young targets. In the second experiment ninety young adults rated the degree to which they considered targets (described in the same vignettes used in the first experiment) to possess specific traits (e.g., responsible) that were both desirable and relevant to performing memory tasks. Nonforgetful targets received higher ratings than forgetful targets and older targets received higher ratings than young targets. The perception that older targets possess such traits to a greater degree than young targets may explain the initial finding that respondents were more likely to choose old over young targets to perform memory tasks. PMID- 8406975 TI - Solar heat load on man. Review of different methods of estimation. AB - Different methods have been compared for the estimation of solar heat load on man. The comparison comprised several methods based on the calculation of absorbed solar radiation and one method for calculation of mean radiant temperature (Mrt). Regression analysis was carried out for predicted values and values calculated for a vertical cylinder, assumed as an analog model of a standing man. Regression of mean skin temperature, measured in 10 subjects exposed to solar radiation under a variety of climatic conditions, on predicted radiant heat load was also analysed. Mean skin temperature correlated best with Mrt, accounting for more than 50% of the variance. The results indicated that three methods provide a realistic estimation of the radiation heat load, whereas some methods show deviations of several hundred per cent. PMID- 8406976 TI - Electromagnetic wave emitting products and "Kikoh" potentiate human leukocyte functions. AB - Tourmaline (electric stone, a type of granite stone), common granite stone, ceramic disks, hot spring water and human palmar energy (called "Kikoh" in Japan and China), all which emit electromagnetic radiation in the far infrared region (wavelength 4-14 microns). These materials were thus examined for effects on human leukocyte activity and on lipid peroxidation of unsaturated fatty acids. It was revealed that these materials significantly increased intracellular calcium ion concentration, phagocytosis, and generation of reactive oxygen species in neutrophils, and the blastogenetic response of lymphocytes to mitogens. Chemotactic activity by neutrophils was also enhanced by exposure to tourmaline and the palm of "Kikohshi" i.e., a person who heals professionally by the laying on of hands. Despite the increase in reactive oxygen species generated by neutrophils, lipid peroxidation from unsaturated fatty acid was markedly inhibited by these four materials. The results suggest that materials emitting electromagnetic radiation in the far infrared range, which are widely used in Japan for cosmetic, therapeutic, and preservative purposes, appear capable of potentiating leukocyte functions without promoting oxidative injury. PMID- 8406977 TI - Low twinning rate and seasonal effects on twinning in a fertile population, the Hutterites. AB - This paper analyzes from the mid 18th century to 1987 the birth records of the "Dariusleut," one of the three subgroups of the Hutterite population. The aim of this study is to describe several aspects of the twinning rate in a fertile population. The overall rate of twinning was 0.90%: 103 twins among all 11,492 maternities. The rate peaked at the 7th birth order and at the maternal age of 40 years and over. Until the mid 19th century when the Hutterites lived in Russia, the twinning rate was higher (1.5%), and it decreased during the migration period in the second half of the 19th century (0.7%). After the group had settled in the USA and Canada, the population maintained a twinning rate of 1.0% until 1965. After 1965 the rate decreased to 0.7%, partly due to a decline in fertility among women aged 30 years and over. There was a significant seasonal variation: the twinning rate decreased to 0.5% in May-July compared to 1.2% for the other three seasons during the years up to 1965 (P < 0.01), while more recent mothers did not show such a seasonal variation. The incidence of twin births in this population seems to have been influenced by environmental factors, which would change their effect seasonally and secularly. PMID- 8406978 TI - Tobacco use and nasopharyngeal carcinoma in a cohort of US veterans. AB - The risk of nasopharyngeal carcinoma (NPC), a relatively rare neoplasm in the United States, was examined in relation to tobacco use in a cohort of nearly 250,000 US veterans whose mortality experience was followed for 26 years. A total of 48 NPC deaths were identified during the follow-up period. Current smokers had a nearly 4-fold increase in risk of NPC compared with non-users of any tobacco, with risk increasing to 6.4 among those smoking more than 2 packs daily. After adjustment for age and number of cigarettes smoked, risks were inversely associated with age at starting to smoke, with the highest risk observed among those who started smoking before age 15, although no clear trend associated with duration of smoking was observed. Former cigarette smokers had a small excess risk of NPC, but no association was detected for cigar/pipe users. This study adds strong evidence to the increasing literature linking cigarette smoking to NPC. PMID- 8406979 TI - Liposarcoma: a population-based epidemiologic and prognostic study of features of 43 patients, including tumor DNA content. AB - Different conceptions exist regarding the epidemiology and prognosis of liposarcoma, and several classification systems are in use. We analyzed a population-based, 25-year series of 43 patients with liposarcoma of the extremity or trunk wall. Follow-up was complete. The annual incidence was 0.12/10(5). The thigh was the most common location. One of 6 tumors was subcutaneous. Deep-seated tumors were larger than s.c. tumors. Among the 42 surgically treated patients, grade II (4-grade scale) was the most common malignancy grade. Four tumors were well-differentiated, 24 were predominantly myxoid, 4 predominantly round-cell, and 10 were predominantly of pleomorphic type. The 5-year metastasis-free survival rate was 69%. By univariate analysis increasing malignancy grade, tumor necrosis, vascular invasion, mitotic count, subtype other than well differentiated, and high cellularity were prognostic for metastatic disease. However, in the multivariate analysis only tumor necrosis was an independent risk factor. Tumor necrosis should be considered when prognosis of liposarcoma of the extremity and trunk wall is evaluated. PMID- 8406980 TI - The value of histoquantitative measurements in prognostic assessment of renal adenocarcinoma. AB - In a series of 135 patients with renal-cell carcinomas (followed up for a mean of 9.5 years), a variety of clinical and histological factors were analyzed in relation to morphometric measurements of the nuclear parameters in the primary tumours to establish their value as prognostic factors. Clinical, histological and morphometric factors were significantly interrelated in that the metastatic high-grade tumours had larger nuclei, larger variation in nuclear size and shape, and were also rapidly proliferating. In a univariate analysis, the most important clinical predictor of recurrence-free survival (RFS) was T category, followed by combined nuclear grade, N category, nuclear grade, tumour size, sex and M category. The most important quantitative predictor of RFS was the mean area of the 10 largest nuclei (NA10), the mean of the longest nuclear axis (Dmax), SD of nuclear area (SDNA), the volume-corrected mitotic index (M/V index), inflammatory cell reaction, SD of nuclear perimetry (SDPE), and the mean of nuclear area (NA). M category, T category, combined nuclear grade, nuclear grade and N category were significantly related to patient survival. Of the quantitative variables, M/V index, Dmax and NA were significant predictors of survival in a univariate analysis. Females had longer RFS than men, and density of tumour-infiltrating lymphocytes (TIL) referred to an increased risk of recurrent tumour in both sexes. In a multivariate analysis, the RFS was independently predicted by the clinical stage, female sex and mitotic frequency/mm2, while nuclear parameters or nuclear grading had no independent prognostic value. The extent of the primary tumour was the single most important determinant of survival, followed by the proliferation rate of the tumour. In local T1-2NOMO tumours, mitotic frequency/mm2 was the only independent prognostic factor for RFS. The clinical stage, mitotic frequency/mm2, nuclear grade and density of TIL were independent predictors in Cox's analysis. In these local tumours, mitotic frequency/mm2 of neoplastic epithelium was the only independent prognostic factor. The results indicate that although an accurate prognostic evaluation of renal-cell carcinomas can be based on subjective nuclear grading and histoquantitative measurements of nuclear parameters, the simple assessment of mitotic frequency alone supplies most of the prognostic data, particularly in local tumours. PMID- 8406981 TI - Cutaneous malignant melanoma: association with height, weight and body-surface area. a prospective study in Norway. AB - To investigate whether anthropometric indices as body height, body-mass index (BMI) and body-surface area (BSA) have any influence on the risk of cutaneous malignant melanoma (CMM), we analyzed height and weight data from a Norwegian screening survey running between 1963 and 1975 and comprising 1.3 million individuals followed for 14 to 26 years. Among females, CMM was diagnosed in 2814 incident cases, while the number among males was 2144 during 22,988,345 person years of follow-up. With adjustment for age, birth cohort, geographic region and BMI, the risk for CMM increased significantly with increasing quintiles of height in both sexes. The tallest persons were at greater relative risk than the shortest ones: 1.60 and 1.59 in males and females respectively. This positive association was also found for CMM localized on face and trunk in both sexes, and in the lower limbs only in females. Males in the highest quintile of BMI had a relative risk for CMM of 1.26 compared with the lowest quintile, while a non significant negative association were found in obese females. This negative association increased and was significant for CMM located on the trunk and lower limbs in females. Among males we observed a consistent association between BSA and risk of CMM similar to that for height. These relationships between anthropometric indices and CMM are discussed according to factors which promote growth, among which nutrition, social class and hormones are the most important. PMID- 8406982 TI - Tumor-cell proliferation and prognosis in renal-cell carcinoma. AB - In the present study the prognostic value of tumor S-phase fraction was evaluated in 69 patients with renal-cell carcinoma. The proportion of S-phase cells was calculated from DNA histograms by flow cytometry in multiple samples from each tumor. The mean tumor S-phase percentage varied between 1.0 and 17.0%, mean 7.5%, with a significant difference between diploid and non-diploid tumors. Stage-I tumors showed significantly lower S-phase values than tumors of stages II/III and IV. Histopathological grade correlated with fraction of cells in S-phase. Nineteen tumors were homogeneously diploid, I was tetraploid and 49 were aneuploid. Heterogeneity concerning S-phase values was found in 46 of the 69 tumors and concerning DNA ploidy in 34 tumors. Survival time was longer for patients with diploid tumors than for those with aneuploid tumors. Patients with S-phase values < 7.5% had a longer survival time than patients having tumors with S-phase values > 7.5%. Within the group of aneuploid tumors, patients with S phase values < 7.5% had a significantly better prognosis. In multivariate analysis, only tumor stage and S-phase gave significant independent prognostic information. The S-phase fraction seems to be an additional prognostic parameter for patients with renal-cell carcinoma. PMID- 8406983 TI - The value of hepatitis B x antigen as a prognostic marker in the development of hepatocellular carcinoma. AB - Chronic infection with hepatitis-B virus (HBV) is associated with high risk for the development of hepatocellular carcinoma (HCC). Several studies have implicated that the X gene product(s) of HBV are important to the pathogenesis of HCC. This study tests the hypothesis that immunohistochemical detection of hepatitis B x antigen (HBxAg) is closely associated with HCC. The patterns of HBxAg were determined by staining in tumor and non-tumor liver sections from 30 Chinese patients with HBV-associated HCC, and the results were compared with other markers of infection. HBxAg was the most prevalent marker of HBV infection both in tumor and in non-tumor tissues of HCC patients, as compared with the hepatitis-B surface and core antigens. This pattern was observed among carriers as well as several patients who were HBsAG- in serum. The HBxAg staining results were validated by Southern blotting with an X-region probe and by Western blotting with anti-HBx. These results suggest that the persistence of HBxAg is important to the pathogenesis of early HCC and that HBxAg expression in the liver during chronic HBV infection may be an important prognostic marker for the development of HCC. PMID- 8406984 TI - Correlation of epithelial proliferation and squamous esophageal histology in 1185 biopsies from Linxian, China. AB - Epithelial proliferation is an active area of research in gastrointestinal cancer, but only a few studies have examined the relationship of esophageal epithelial proliferation and squamous histologic findings in populations with high rates of squamous esophageal cancer. In order to study this correlation, tritiated thymidine labeling was performed on 1185 esophageal biopsies from 745 residents of Linxian, China, a county with some of the highest esophageal-cancer rates in the world. Total labeling index (TLI = total labeled cells/total cells counted) was used to measure the amount of proliferation, and the proportion of labeled cells found in cell layers 4 to 10 (labeled cell fraction 4 plus, LF4+ = labeled cells in layers 4-10/total labeled cells) was used to measure the vertical distribution of proliferation. Of the biopsies, 979 were histologically normal, 51 showed acanthosis, 35 showed esophagitis, 116 showed squamous dysplasia, and 6 showed invasive squamous cancer. The mean values of both proliferation variables, stratified by histologic diagnosis, showed the following relationships: normal = acanthosis < esophagitis = dysplasia < cancer. The ranges of proliferation values overlapped extensively in all biopsy categories, so that measuring proliferation could not substitute for histologic diagnosis. It remains to be seen whether proliferation values, histologic diagnoses, or some combination of these methods is most predictive of subsequent esophageal cancer. PMID- 8406985 TI - E-cadherin expression in head and neck squamous-cell carcinoma is associated with clinical outcome. AB - The cell-cell adhesion molecule E-cadherin has been shown to suppress invasive growth of epithelial cells in vitro, and loss of its expression is thought to be important in invasion and metastatic potential of epithelial tumors in vivo. We retrospectively studied the level of E-cadherin expression in 50 primary head and neck squamous-cell carcinomas (HNSCC) by immunohistochemical methods, on frozen sections, using anti-E-cadherin monoclonal antibody (MAb) 6F9. It concerned patients with different stages of carcinoma of larynx or oral cavity who had been treated with curative intention 30 months or more before. Percentages of membranous stained tumor cells were scored in 1 of 5 categories. Scores were generally low, as in 11/50 lesions < or = 5% cells were stained, and in 19/50 lesions only 6-25% cells showed membranous staining. In 9 lymph-node metastases evaluated, E-cadherin expression was in the same range as in the primary tumors. There was a significant correlation between the level of membranous E-cadherin expression in the primary tumor and the degree of differentiation. No relation was found with tumor size (pT) or regional lymph-node classification (pN). Nevertheless, 29 patients surviving > or = 30 months without evidence of disease had significantly higher levels of membranous E-cadherin expression in their primary tumors than 10 patients with unfavorable clinical course clearly related to recurrent and/or metastatic HNSCC. Moreover, this could only partially be explained by distinctions in differentiation grade between both groups. Our results suggest that membranous E-cadherin expression has prognostic importance in patients with HNSCC. PMID- 8406986 TI - Possible regulation of soluble ICAM-1 levels by interleukin-1 in a sub-set of breast cysts. AB - There are 2 main groups of breast cysts characterized by their intracystic sodium to-potassium ratios. Women who have intracystic Na/K < 3 may have a higher risk of developing breast cancer than those who have intracystic Na/K > 3. The cell adhesion molecule, ICAM-I, has been shown to be inducible by interleukin-I (IL I). Furthermore, ICAM-I may be involved in the progression of metastasis in certain malignancies. The aim of this study was to measure intracystic soluble ICAM-I (sICAM-I) and IL-I alpha/beta levels and to assess the relationships between these substances. Wide-ranging concentrations of sICAM-I, IL-I alpha and IL-I beta were found in breast-cyst fluid. The high-electrolyte-ratio group had intracystic levels of sICAM-I and IL-I beta which were significantly higher than those of the low-electrolyte-ratio group. No significant difference was found between IL-I alpha concentrations in the 2 groups of breast cysts. Strong positive correlations were found between sICAM-I and IL-I alpha, sICAM-I and IL-I beta, and IL-I alpha and IL-I beta. The significance of the higher concentrations of sICAM-I in the high-electrolyte-ratio group remains to be determined, but the results of this study suggest that sICAM-I levels in the high-electrolyte-ratio group may be regulated in part by IL-I alpha and IL-I beta. PMID- 8406987 TI - Abnormalities affecting the APC and MCC tumour suppressor gene loci on chromosome 5q occur frequently in gastric cancer but not in pancreatic cancer. AB - Abnormalities affecting tumour suppressor genes on chromosome 5q21 are increasingly recognised as important in the pathogenesis of a variety of human cancers, particularly of the gastrointestinal tract. We have examined a series of gastric and pancreatic cancers from European patients for loss of heterozygosity (LOH) of markers within and around the APC and MCC genes on chromosome 5q21 using restriction fragment length polymorphism and polymerase chain reaction techniques. We find that LOH of the APC and MCC genes is particularly frequent in gastric cancers of diffuse type, but very infrequent in pancreatic cancers. We have also used single-strand conformational polymorphism to screen for abnormalities of the sequence of the APC and MCC genes in a panel of pancreatic cancer cell lines. Our results suggest that there are distinct differences in the molecular pathogenesis of gastric and pancreatic cancer and that abnormalities of APC and MCC may be involved particularly in the diffuse type of gastric cancer. PMID- 8406988 TI - PCR detection and typing of genital papillomavirus in a New Brunswick population. AB - We have used a broad range of primers for HPV detection, using the polymerase chain reaction (PCR) so as to compare PCR typing of HPV with the results of cytological diagnosis in a New Brunswick population referred to the out-patient clinic of the Saint John Regional Hospital. The primers selected were found to be capable of amplification with high efficiency, therefore we did not perform further hybridization analysis for specific identification of HPV types. Amplification of selected fragments for detection of HPV 6, 11, 16, 18, 31 and 33 was obtained from cervical swabs collected from 154 patients. Microscopic examination was performed in duplicate samples and the results compared with the DNA-typing analysis. HPV of any of the above types was detected in 43 out of 154 patients. Among these, 32 patients showed single or multiple infections with "high-risk" HPV strains 16, 18, 31 or 33. Cytologically normal or atypical samples with any of the HPV types tested amounted to 17%, but increased to 56% in patients with CIN I, and to 100% in patients with CIN II or III. Prevalence of "high-risk" types alone increased from 15% and 10%, for normal and atypical cases respectively, to 48% for CIN I, 75% for CIN II and 100% for CIN III. Our results indicate that HPV detection and typing by this simple procedure can be a valuable indicator of cancer progression and thus can help to identify individuals at high risk in pre-malignant stages of the disease. PMID- 8406989 TI - Expression of Epstein-Barr-virus-related nuclear antigens and B-cell markers in lymphomas of SIV-immunosuppressed monkeys. AB - Simian-immunodeficiency-virus(SIV)-infected cynomolgus monkeys develop B-cell lymphomas in approximately one third of the cases. We have now studied the expression of cynomolgus-Epstein-Barr-virus(cyno-EBV) nuclear antigens in 13 cyno EBV-carrying SIVsm-associated monkey lymphomas and established cell lines from 3 of these tumors. Immunoblots of cell lysates were probed with polyspecific and monospecific reagents directed against human EB-virus EBNAI-6, and against the membrane protein LMPI. An EBNA2-cross-reacting protein was demonstrated in 8 lymphoma tissues (8/13) and in the 3 cell lines derived from the tumors. All tumors expressed a polypeptide with 50 to 55 kDa molecular weight, which cross reacted with some antibodies to EBNAI. Absorption experiments with normal monkey tissue showed that this polypeptide was specific for the cyno-EBV-carrying lymphoma cells. Equivalents of EBNA3-6 and LMPI could not be detected. Immunophenotypical characterization showed that the monkey lymphomas were similar to human HIV-associated B-cell lymphomas. Malignant B-cell lymphomas in experimentally SIVsm-infected cynomolgus monkeys can be a model for EBV associated lymphomagenesis in immunodeficiency states. PMID- 8406990 TI - Cis-diamminedichloroplatinum(II) inhibits p34cdc2 protein kinase in human lung cancer cells. AB - cis-Diamminedichloroplatinum(II) (CDDP) induced G2-phase arrest in PC-9 human cancer cells. To elucidate how CDDP acts on cell-cycle regulation, we analyzed the effect of CDDP on cell-cycle regulators such as p34cdc2 protein kinase. p34cdc2 protein kinase activity was maximum in G2 phase and decreased after G2/M transition in synchronized PC-9 human lung cancer cells. Evidence for a phosphorylated p34cdc2 protein kinase complexed with cyclin B was obtained from cells in G2 phase and the p34cdc2 protein kinase appeared to be dephosphorylated at M phase. After exposure to CDDP in G1 phase, PC-9 cells were arrested in G2 phase. The activation of p34cdc2 protein kinase was inhibited by CDDP. Cyclin A and wee-I kinase were not affected by the exposure to CDDP. Cyclin B was degraded in M phase in PC-9 cells. Exposure to CDDP did not affect the degradation of cyclin B. Our data suggest that the effect of CDDP on cell-cycle phase might be regulated by the dephosphorylation of p34cdc2 protein kinase. To determine whether the p34cdc2 protein kinase is a primary target for CDDP, we examined the direct effect of CDDP on tyrosine dephosphorylation of p34cdc2 protein kinase in cellular extracts. Cell lysates from synchronized PC-9 in G2 phase were immunoprecipitated with p13-Sepharose beads. In vitro dephosphorylation of phosphotyrosine of p34cdc2 protein kinase was observed after exposure to okadaic acid in a concentration-dependent manner. The dephosphorylation of p34cdc2 protein kinase by okadaic acid was inhibited by CDDP. We hypothesize that inhibition of p34cdc2 dephorphorylation by CDDP is important for its growth inhibiting properties. PMID- 8406991 TI - Moloney virus induction of T-cell lymphomas in a plasmacytomagenic strain of E mu v-abl transgenic mice. AB - Although the v-abl gene can provoke several types of lymphoid neoplasm, mice of a transgenic strain (E mu-v-abl 40) in which lymphocytes are targeted for expression of v-abl by a linked immunoglobulin enhancer (E mu) spontaneously develop only plasmacytomas. To determine whether other lymphocytes of this strain were susceptible to transformation, and to identify genes that can collaborate with v-abl in tumorigenesis, E mu-v-abl 40 mice were subjected to insertional mutagenesis by neonatal infection with Moloney murine leukemia virus. Tumorigenesis was accelerated moderately, but nearly all the tumors were T lymphomas. The altered tumor type may reflect both the T-cell tropism of Moloney virus and the higher level of E mu-v-abl 40 expression found in T lymphocytes than in B lymphocytes. Insertion near the c-myc, N-myc or pim-I gene was observed in 42% of the induced tumors, indicating that each of these genes may collaborate with v-abl in lymphomagenesis. Most of the accelerated tumors had a surprisingly low level of transgene expression. Thus, high expression of v-abl may not be required for Moloney-induced T lymphomagenesis. PMID- 8406992 TI - Flunarizine as a modulator of doxorubicin resistance in human colon adenocarcinoma cells. AB - The potential of the calcium-entry blocker flunarizine in modulating the cytotoxicity of doxorubicin was investigated in human colon-adenocarcinoma cell lines sensitive to (LoVo) or with experimentally induced resistance (LoVo/DX) to doxorubicin. Exposure to 1 to 2 micrograms/ml flunarizine for intervals of up to 24 hr did not affect cell survival in either line. Simultaneous exposure to flunarizine and doxorubicin for 1 hr selectively enhanced doxorubicin activity in the resistant cell line and not in the sensitive cell line. In particular, the doxorubicin concentration able to reduce cell survival by 50% dropped to one third. Moreover, simultaneous exposure to flunarizine significantly increased intracellular doxorubicin accumulation, as evaluated by fluorescence spectrophotometry. Again, flow-cytometric analysis showed hyperpolarization of the membrane in resistant cells, starting from 15 min of exposure to 2 micrograms/ml flunarizine. Finally, in LoVo/DX cells, which normally express gp170, a 24-hr treatment with flunarizine markedly reduced the immunoreactivity of cells with 2 monoclonal antibodies (MAb57 and MRK16) directed against different external epitopes of the glycoprotein. The results from our study indicate the ability of flunarizine to positively modulate doxorubicin-resistance in human colon-adenocarcinoma cells expressing the multidrug-resistance phenotype. PMID- 8406993 TI - Importance of mevalonate-derived products in the control of HMG-CoA reductase activity and growth of human lung adenocarcinoma cell line A549. AB - HMG-CoA reductase catalyzes the synthesis of mevalonate, a crucial intermediate in the biosynthesis of cholesterol and non-sterol isoprenoid compounds essential for cell growth. The HMG-CoA reductase activity of the A549 tumor cell line is higher than that of normal human fibroblasts. This deregulation in mevalonate needs was not due to an alteration in the activated state of the enzyme by short term regulation. We show that the HMG-CoA reductase in A549 cell line was subject to a multivalent feedback control. A high fraction (40%) of the reductase activity was devoted to non-sterol products. In contrast, normal fibroblasts had only 15-20% of the reductase activity that generated non-sterol products. We also show that cholesterol and at least one of the non-sterol products are necessary for optimal cell growth of A549 cells. Our data strongly suggest that A549 cells produce more non-sterol substances which may be related to increased requirements of mevalonate for upregulated cell growth. PMID- 8406994 TI - Induction and release of manganese superoxide dismutase from mitochondria of human umbilical vein endothelial cells by tumor necrosis factor-alpha and interleukin-1 alpha. AB - The effects of tumor necrosis factor-alpha (TNF alpha) on cultured human umbilical vein endothelial cells (EC) and 2 cancer cell lines, A549 and ME180, were compared. The effects of interleukin-1 alpha (IL-1 alpha) on EC were also examined. While A549 cells were fairly resistant to the cytolytic effects of TNF alpha and IL-1 alpha, ME180 cells were sensitive. EC were also less sensitive to TNF alpha than ME180 cells, as judged by viability of individual cells and by the release of lactate dehydrogenase (LDH) into the medium. Both manganese superoxide dismutase (Mn-SOD) and its mRNA were markedly induced by these cytokines in EC and in A549 cells but not in ME180 cells. The levels of Mn-SOD in the conditioned medium of EC were markedly increased after stimulation with cytokines, whereas those in ME180 and A549 cells were relatively low. The amount of Mn-SOD released appears to be comparable to that from cells lysed due to the cytocidal effect of cytokines, as assessed by measuring intra- and extra-cellular LDH activity. These data suggest that, in vivo, the TNF alpha and IL-1 alpha produced by cancer cells and other cells may induce Mn-SOD in vascular endothelial cells as well as other host tissues, resulting in release of a relatively large amount of this protein into the serum. PMID- 8406995 TI - Production of monoclonal antibodies to human estrogen-receptor protein (ER) using recombinant ER (RER). AB - Two monoclonal antibodies (MAbs), IC5 and ID5, were produced using spleen cells from BALB/c mice immunized with recombinant estrogen-receptor protein (RER). On immunoblotting, both MAbs reacted with the 67-kDa polypeptide chain obtained by transformation of E. coli and transfection of COS cells with plasmid vectors expressing ER. The epitopes of both MAbs were in the N-terminal domain (A/B region) of the receptor. In normal human tissues, IC5 and ID5 reacted with cells known to contain large amount of ER, such as cells of the mammary gland and the uterus. Staining was localized predominantly in nuclei with little or no cytoplasmic reactivity. IC5 and ID5 were unreactive with tissues usually considered to be negative for ER. The reactions of these 2 MAbs were further tested on different tumor types, using immunohistochemical (IHC) method on frozen sections. In breast cancer, a good correlation was found between the results obtained on frozen sections and those using the conventional radioligand dextran coated charcoal (DCC) assay. Immunostaining with IC5 and ID5 MAbs was also assessed on routinely processed paraffin sections using the antigen-retrieval method. Staining was comparable to that obtained on frozen sections in virtually all the breast carcinomas. Negative reactions were consistently obtained with both antibodies on human neoplasms derived from other non-estrogen-dependent organs. IC5 and ID5 MAbs may thus be of value in routine diagnostic histopathology for assessment of the estrogen-receptor content in human carcinomas. PMID- 8406996 TI - A sub-set of immediate early mRNAs induced by tumor necrosis factor-alpha during cellular cytotoxic and non-cytotoxic responses. AB - TNF-alpha is a multifunctional cytokine which is cytotoxic for some cell lines. In order to characterize the early genomic response to TNF-alpha, we have analyzed the induction of a sub-set of serum-inducible immediate early genes in WEHI-S and L929 fibrosarcoma cell lines, which are sensitive to TNF-alpha, and in the 3T3-LI pre-adipocytic cell line, which is resistant to TNF-alpha cytotoxicity. Among 77 immediate early mRNAs screened by dot blot and/or Northern blot analyses, the expression of 23 mRNAs was found to be induced by TNF-alpha. Ten of these mRNAs encode proteins known to function as pro-inflammatory cytokines or transcription factors, while 13 others have as yet uncharacterized activities. The magnitude of c-fos induction by TNF-alpha inversely correlated with cell-type-specific cytotoxicity. Rapid and transient mRNA responses were observed in the TNF-alpha-resistant cells, whereas a slower and more persistent response was characteristic for TNF-alpha-sensitive cells. The prolonged induction of immediate early mRNAs may contribute to TNF-alpha-induced cellular cytotoxic responses. PMID- 8406997 TI - Role of cell cholesterol in modulating vincristine uptake and resistance. AB - The relationship between cell-membrane permeability to vincristine and cholesterol/phospholipid levels was studied in L5178Y murine leukemic lymphoblasts and in 2 multidrug-resistant cell sublines, VCR/P60 and VCR/P200, which expressed increasing levels of vincristine resistance. The uptake of 3H vincristine was measured in all cell lines and in cholesterol-depleted and reloaded L5178Y and VCR/P200 cells. The initial rate of drug entry in resistant cells was lower than that measured in the parental cell line and it decreased as the relative resistance increased. An increment of cholesterol content, characterized in resistant cells, was directly proportional to the relative resistance to vincristine. Cholesterol depletion in both sensitive and resistant cells resulted in an increase in the rate of vincristine uptake, which reverted to the respective basal levels when each cell line was cholesterol-reloaded. The rate of drug uptake was inversely correlated with the molar ratio of cholesterol to phospholipids. Although both VCR/P cell sublines, but not the sensitive parental cells, expressed the P-glycoprotein in their plasma membrane, there were no differences in drug efflux and retention between resistant and parental cells. These results indicate that cholesterol modulates the permeation of vincristine through the plasma membrane and strongly suggest that increased levels of cholesterol/phospholipid account for the lower drug accumulation and greater resistance in these multidrug-resistant cells. PMID- 8406998 TI - Parvovirus H-1 inhibits growth of short-term tumor-derived but not normal mammary tissue cultures. AB - Infection with parvovirus H-1 strongly interfered with the proliferation of non established tissue cultures derived from human breast tumors, but had little effect on the growth of corresponding normal human mammary cells. Even though tumor cells were always more sensitive to the virus than normal tissue from the same patient, appreciable quantitative differences were observed among tumor specimens. With time and sub-cultures, the killing effect of the virus on tumor cells became amplified. The impaired growth of infected tumor cells was due both to cytotoxic and to cytostatic action of H-1 virus and was associated with their greater capacity for virus-DNA amplification as compared with normal cells. PMID- 8406999 TI - Increased accumulation of p53 protein in cisplatin-resistant ovarian cell lines. AB - We have examined p53 protein levels in cell lines selected for resistance to the chemotherapeutic drug cis-diamminedichloroplatinum (II), cisplatin. The majority of the independent cisplatin-resistant clones isolated by a single selection with cisplatin from the ovarian tumour cell line A2780 showed increased levels of p53 protein compared to the parental cell line. Elevated p53 protein levels were also observed in cisplatin-resistant ovarian human tumour lines isolated after multiple exposures to cisplatin (A2780/cp70 and OVIP/DDP). Direct PCR sequencing of p53 cDNAs showed that both the A2780/cp70 and the parental A2780 cell lines had a wild-type p53 gene sequence. The OVIP and OVIP/DDP lines both had a heterozygous mutation at codon 126. Cell-cycle analysis after gamma-irradiation or cisplatin treatment showed evidence of a G1/S and G2/M cell-cycle checkpoint in both A2780/cp70 and the sensitive parental cell lines. However, the resistant cell line A2780/cp70 showed less inhibition of DNA synthesis after gamma irradiation than the sensitive cell line. Transfection of a mutant p53 gene construct (containing a mutation at codon 143, val to ala) into the A2780/cp70 resistant cells conferred a significantly increased sensitivity to cisplatin, suggesting that p53 is a direct determinant of cisplatin resistance in these cells. However, expression of this mutant p53 in the A2780 cells did not affect sensitivity. PMID- 8407000 TI - Mitotic activity, apoptosis and TRPM-2 mRNA expression in DMBA-induced rat mammary carcinoma treated with anti-estrogen toremifene. AB - Anti-estrogen toremifene inhibits growth of the 7,12 dimethylbenz(a)anthracene(DMBA)-induced rat mammary carcinoma. The changes in proliferation and cell death were studied in detail. Proliferation was measured by counting mitotic figures in histologic sections, expressed by volume-corrected mitotic index (M/V INDEX). Respective volume corrected apoptotic index (A/V INDEX) was measured by counting apoptotic nuclei in the same sections. The presence of apoptotic cells was confirmed by transmission electron microscopy. In the untreated tumors, both mitosis and apoptosis were frequent. In the toremifene treated tumors, the mean M/V INDEX was about half of the mean M/V INDEX in the untreated control tumors. The mean A/V INDEX in the toremifene-treated tumors was about 3/4 of the mean A/V INDEX in the controls. In toremifene-treated tumors, A/V INDEX was strongly correlated with TRPM-2-gene expression, which was also enhanced when compared with the controls. TRPM-2 gene has been associated with programmed cell death induced by hormonal ablation in prostate- and breast-cancer cells. No such correlation was seen in control tumors. These findings suggest that, in the DMBA-tumor model, toremifene affects the cell turnover by inhibiting mitotic activity and modifying abundant spontaneous apoptosis. In this model, the inhibition of tumor growth results mostly from reduction of mitotic activity. PMID- 8407001 TI - Nerve growth factor effects on human and mouse melanoma cell invasion and heparanase production. AB - The role of growth factor networks in regulating the progression of human melanocytes towards tumorigenicity and ultimately the malignant phenotype is poorly understood. In particular, the autocrine and paracrine influences that modulate cellular invasion and extracellular matrix degradative enzymes of melanoma cells remain undefined at the molecular level. We report here that nerve growth factor (NGF) can modify some metastasis-associated cellular properties of human and mouse melanoma cells. Treatment of early-passage human metastatic melanoma cells (MeWo) or their variants (3S5, 70W) with biologically active 2.5S NGF resulted in (a) delayed density-dependent inhibition of melanoma cell growth; (b) increased in vitro invasion through a reconstituted basement membrane; and (c) time- and dose-dependent induction of heparanase, a heparan-sulfate-specific endo-beta-D-glucuronidase associated with human melanoma metastasis. These effects of NGF were most marked in the 70W brain-colonizing cells (70W > MeWo > 3S5). The NGF enhancement of heparanase secretion was not species-specific, since it was also observed in murine B16 melanoma cells; the highest NGF stimulation of heparanase was found in brain-colonizing murine B16-B15b variant (B16-B15b > B16 BL6, B16-F10, B16-F1). NGF also increased the invasive capacity of the human 70W and murine B16-B15b sublines in a chemoinvasion assay performed with filters coated with purified heparan sulfate proteoglycan (HSPG). The enhancement of chemotactic response and heparanase production was detected at NGF concentrations sufficient to fully saturate both low- and high-affinity NGF receptors (NGFR), the neurotrophin receptor (p75) and the trkA gene product, respectively. The results suggest that, in addition to the effects of NGF on cellular development and differentiation within the peripheral and central nervous systems, NGF can exert changes in the invasive properties of neuroectoderm-derived melanoma cells. PMID- 8407002 TI - Immunohistochemical analysis of ras p21 expression in normal, pre-malignant and malignant oral mucosa. PMID- 8407003 TI - Dietary electrolytes and urinary natriuretic factors. AB - We examined the relationship between the excretion of electrolytes (sodium, potassium and calcium), dopamine and digoxin-like immunoreactive substance in 41 young healthy female subjects (age 18-23 years) in order to study the interaction of electrolyte intake on dopamine and digoxin-like immunoreactive substance- factors which have been postulated to have a pathogenic role in hypertension. Sodium excretion was significantly correlated with dopamine excretion (r = 0.545, P < 0.0005) and digoxin-like immunoreactive substance (r = 0.359, P < 0.02). There was also a significant correlation between calcium and digoxin-like immunoreactive substance (r = 0.345, P < 0.03). Stepwise multiple regression analysis further confirmed that sodium is the only contributor to dopamine excretion and calcium is the only contributor to digoxin-like immunoreactive substance (r2 = 0.114). We conclude that in young healthy subjects dopamine excretion is determined partly by sodium intake and that the excretion of digoxin like immunoreactive substance is independent of sodium intake. PMID- 8407004 TI - Prevalence of coronary heart disease in Turkish adults. AB - The prevalence of coronary heart disease was determined by a conducted survey in a random sample of 3689 subjects 20 years of age or older in 59 communities representing the Turkish adult population. Interview with a questionnaire, physical examination of the cardiovascular system and recording of a 12-lead ECG were performed. The latter was coded according to the Minnesota code. Expressed in age-adjusted rates (for 35-64 years), prevalence rates per 100 men were as follows: typical angina 3.7, atypical angina 0.9, electrocardiographic evidence of myocardial infarction and/or ischemia 3.7, any of the stated findings suggesting coronary heart disease 8. Women had a substantially higher rate of atypical angina, positive ECG findings and of any of the stated manifestations for coronary heart disease, whereas they had a significantly lower rate of Q/QS patterns as well as of a history of myocardial infarction. Based on a probability related point score, age-adjusted clinical coronary heart disease was estimated to prevail in 5.8% of men and 5% of women (P > 0.4) in the sample of the Turkish population. The respective rates in urban residents was 6% and in rural resident 4.8%. Among participants diagnosed coronary heart disease, 63% presented the form of angina without infarction, 27% had evidence of myocardial infarction, 7% 'silent myocardial ischemia' and 3% cardiac failure alone. PMID- 8407005 TI - Prognosis after a first Q-wave myocardial infarction in ethnic Chinese people: a prospective study. AB - The prognosis after a first Q-wave myocardial infarction was investigated in 206 Chinese patients of 65 years or younger who had a predischarge cardiac catheterization and coronary angiography. Three patients studied were lost to follow-up. In the remaining 203 patients with ages of between 28 and 65 years, 101 (49.8%) had 0- or 1-vessel disease, 56 (27.6%) had 2-, and 38 (18.7%) had 3 vessel disease. Significant left main coronary artery stenosis was noted in 8 (3.9%). During a mean follow-up of 33 months, 33 (16.3%) patients had 36 episodes of cardiac events, and 16 (7.9%) died of cardiac causes. Stepwise logistic regression analysis revealed that the left ventricular ejection fraction and left main coronary artery disease were predictors of cardiac mortality, while age and the extent of coronary artery disease were predictors of total cardiac events. There was no variable that could predict recurrence of myocardial infarction. PMID- 8407006 TI - Cardioprotective effect of intracoronary nifedipine during percutaneous transluminal coronary angioplasty. A French double-blind cross-over multicentre study. AB - The aim of this double-blind, placebo-controlled, cross-over study was to assess the cardioprotective effect of intracoronary nifedipine during percutaneous transluminal coronary angioplasty balloon occlusion. A balloon inflation without drug injection was initially made to ascertain that a shift of the ST segment (> or = 2 mm, 0.08 s after the J point) appeared (inclusion criterion). Two other balloon inflations were preceded by intracoronary injection of either 0.2 mg nifedipine or placebo, distal to the stenosis through the balloon catheter. The evaluation criteria were (1) time to ST segment shift, and (2) maximal amplitude of ST segment shift caused by balloon occlusion. Comparison of the data used an analysis of variance. Sixty-seven patients (mean age 54 +/- 8 years; 54 male, 13 female) were studied; 50 patients had 1-, 16 patients 2- and 1 patient 3-vessel disease. The dilated vessel was the left anterior descending coronary artery (n = 51), the right coronary artery (n = 12) and the left circumflex coronary artery (n = 4). Balloon inflation time was 100 +/- 31 s in the nifedipine group and 93 +/- 29 s in the placebo group. Five patients were excluded (procedure stopped after the first inflation in 1 and ST segment shift < 2 mm during the first inflation in 4). The time to 2-mm ST segment shift was longer in the nifedipine group than in the placebo group (62 +/- 40 s versus 51 +/- 40 s, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407007 TI - Somatic and social prognosis of patients with angina pectoris and normal coronary arteriography: a follow-up study. AB - A follow-up over a 7-year period demonstrated that 8.6% of all patients subjected to coronary arteriography because of angina pectoris had normal coronary arteries (NCA). The somatic and social prognosis of these patients were evaluated and these were compared to that of an age- and sex-matched group of patients with arteriographically verified coronary atherosclerosis (CAD). On average 44 months after coronary arteriography, 2.4% with NCA had died versus 20.5% with CAD (P < 0.001). Myocardial infarction occurred in 0% (NCA) versus 12.8% (CAD) among survivors (P < 0.001). Coronary revascularization was carried out in 0% (NCA) versus 76.9% (CAD). Chest pain was unchanged or had worsened in 58.2% (NCA) versus 21.1% (CAD) (P < 0.001) and this in the NCA patients was correlated to the occurrence of minimal lesions of the coronary arteries. Of the NCA patients, 33.3% had ischaemia during exercise-ECG. Normalization was seen in 12 patients and newly developed ischaemia in seven patients. Three patients developed ischaemia during hyperventilation test. Eighty percent (NCA) versus 63.9% (CAD) gave up work due to chest pain (P < 0.001). Further, 55.7% (NCA) versus 34.6% (CAD) had reduced daily activities (P < 0.001); similarly, the frequency of divorce was higher in the NCA group (10.2%) than in the CAD group (1.3%) (P < 0.05). PMID- 8407008 TI - Importance of non-stenosed donor coronary arteries for collateral flow reserve. AB - To elucidate whether the presence of significant stenoses in donor coronary arteries compromises collateral flow reserve, coronary arteriography was performed in 22 patients with stable effort angina who had a complete obstruction in one of the major coronary arteries. The patients were divided into two groups according to the absence (group I: n = 11) or presence (group II: n = 11) of significant stenoses (> or = 75% stenosis of the luminal diameter) in donor arteries. After conventional cardiac catheterization, coronary arteriography was repeated before and during rapid atrial pacing. Coronary collateral circulation was evaluated by means of angiographic contrast appearance time (CAT) which was defined as the time (s) until the epicardial segments distal to the site of complete occlusion were opacified after the injection of contrast medium into the donor artery. The pressure gradient across the collateral circulation (mean aortic pressure minus left ventricular end-diastolic pressure) remained unchanged before and during rapid atrial pacing (group I: 95 +/- 14 vs. 99 +/- 15 mmHg; group II: 91 +/- 18 vs. 94 +/- 16 mmHg). Rapid atrial pacing decreased the contrast appearance time in group I from 1.73 +/- 0.34 to 1.47 +/- 0.37 s (P < 0.01), but was not changed in group II from 1.68 +/- 0.36 to 1.73 +/- 0.51 s (P = n.s.). It was concluded that coronary collateral circulation is further augmented in response to an increase in myocardial oxygen demand in patients with non stenosed donor coronary arteries and significant stenoses in donor arteries compromise an increase in collateral blood flow. PMID- 8407009 TI - Prevalence of coronary heart disease in the rural population of Thiruvananthapuram district, Kerala, India. AB - To establish the prevalence, with 95% confidence limits, of some of the indicators of coronary heart disease in the rural population of Thiruvananthapuram district, Kerala state, India, we did a field survey on a cluster sample with probability proportionate to size (PPS sample) of 500 households from five villages. Altogether the sample consisted of 1253 individuals who were more than 25 years of age, of which 1130 responded (90%). The survey instruments included the Malayalam translation of the Rose questionnaire, a standard 12-lead electrocardiogram with a battery operated portable electrocardiograph machine, blood pressure measurements using a mercury sphygmomanometer, and routine anthropometric measurements. The prevalence rates estimated were: (a) ECG changes suggestive of coronary heart disease, 36/1000 (95% C.L., 18, 55), (b) Rose questionnaire angina, 48/1000 (95% C.L. 35, 62), (c) definitive evidence of coronary heart disease, 14/1000 (95% C.L., 7, 21), (d) possible evidence of coronary heart disease, 74/1000 (95% C.L., 55, 93). Prevalence of major risk factors were, (a) hypertension by the WHO criteria, 179/1000 (95% C.L., 137, 221), (b) smoking, 219/1000 (95% C.L., 151, 287), (c) diabetes, 40/1000 (95% C.L., 17, 63), (d) obesity, 55/1000 (95% C.L., 6, 104). We have found that objective criteria indicate a lower prevalence of coronary heart disease in rural Thiruvananthapuram district when compared to studies from urban centres in India, but the prevalence of angina by Rose questionnaire is greater. PMID- 8407010 TI - Prognostic significance of exercise thallium-201 myocardial perfusion imaging compared to stress echocardiography and clinical variables in patients with unstable angina who respond to medical treatment. AB - The prognostic value of thallium-201 imaging in patients with unstable angina is not well established. Forty consecutive patients with unstable angina who had responded to medical therapy underwent predischarge symptom-limited exercise testing and 39 of them underwent exercise thallium-201 imaging, on average 3 days after the exercise test. Exercise echocardiography was performed in 36 of these patients in conjunction with the predischarge exercise test. Patients with previous myocardial infarction, coronary revascularization, left bundle branch block and dilated cardiomyopathy were not included in the study. An echocardiographic wall-motion score index was derived by analyzing left ventricular regional wall motion. During a follow-up period of 30 +/- 6.4 months, 3 patients had a non-fatal myocardial infarction and 20 required revascularization because of a recurrence of severe medically refractory angina. Univariate predictors of cardiac events (non-fatal myocardial infarction or a need for revascularization) during follow-up included ST-depression during exercise, positive exercise echocardiography, a low exercise wall-motion score index, the presence of thallium-201 redistribution and the number of myocardial segments with thallium-201 redistribution. However, stepwise logistic regression analysis revealed that the presence of thallium-201 redistribution was the only significant non-invasive predictor (P < 0.005) of a cardiac event among patients who underwent predischarge exercise testing and exercise thallium-201 imaging. Among patients undergoing exercise echocardiography and exercise thallium-201 imaging, the number of segments with thallium-201 redistribution was the only significant predictor (P < 0.0005) of future cardiac events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407012 TI - A case report of double-orifice tricuspid valve. AB - A heart specimen with double-orifice tricuspid valve is described. The tricuspid valve was divided into the anterior and posterior orifices by a bridge of leaflet tissue. The valve was stenotic because of the arcade deformity of the anterior papillary muscle to which the bridging leaflet tissue had short chordal insertions. The anterior orifice was regurgitant as a result of a deficient septal leaflet. Clinical correlation was obtained by magnetic resonance imaging of the specimen. The functional significance of the malformed valve was assessed by three-dimensional reconstruction of the two-dimensional magnetic resonance images. PMID- 8407011 TI - Non-specific aortoarteritis: long-term follow-up on immunosuppressive therapy. AB - Thirteen patients with non-specific aortoarteritis and endomyocardial biopsy evidence of myocarditis were followed-up on immunosuppressive therapy comprising of prednisolone and cyclophosphamide in addition to conventional treatment for hypertension and/or congestive heart failure. Serial determinations of erythrocyte sedimentation rate, chest roentgenogram, radionuclide ventriculogram and hemodynamic study including endomyocardial biopsy were carried out at 12, 24 and 52 weeks of therapy. Arterial lesions were also assessed by digital subtraction angiography at 0 and 52 weeks of immunosuppressive therapy. At the end of a year of treatment all patients with congestive heart failure (10/13) showed symptomatic improvement by at least one New York Heart Association (NYHA) class. There was a significant fall in erythrocyte sedimentation rate (48 +/- 12 mm/1st h to 31 +/- 12 mm/1st h, P < 0.05), pulmonary artery pressure (32 +/- 14 mmHg to 20 +/- 9 mmHg, P < 0.05), left ventricular filling pressure (20 +/- 11 mmHg to 11 +/- 7 mmHg, P < 0.05) and increase in left ventricle ejection fraction (39 +/- 16% to 51 +/- 14%, P < 0.05) associated with resolution of morphological changes on endomyocardial biopsy. Arterial lesions remained static with neither progression nor appearance of new lesions. No significant complications of therapy were noticed in any patient. Our uncontrolled observations suggest that immunosuppressive therapy is safe and results in clinical, hemodynamic and myocardial morphological improvement in a subset of patients with non-specific aortoarteritis and associated myocarditis. PMID- 8407013 TI - 1:1 atrioventricular conduction in atrial flutter with digoxin and flecainide. AB - A 65-year-old man with paroxysmal atrial flutter was treated with digoxin and flecainide. The atrial flutter rate was slowed to 190 beats per minute but at times 1:1 AV conduction occurred. PMID- 8407014 TI - Expanded indication: diagnosis of pulmonary embolism by transesophageal echocardiography. AB - Transesophageal echocardiography is a useful diagnostic tool not only in examination of the heart, but also in examination of the great arteries. Its role in the diagnosis of aortic aneurysm and dissection has been well-established. The pulmonary artery, especially the right pulmonary artery, also can be visualized clearly by this technique. In this report, we described a patient with massive pulmonary embolism which was diagnosed by transesophageal echocardiography, and was confirmed by pulmonary angiography. PMID- 8407015 TI - A question of filling in the gaps: a master class commentary. PMID- 8407016 TI - Use of hypnosis following training in a psychiatry residency and psychology internship program: a brief communication. AB - Despite growing numbers of internships and residencies offering training in hypnotherapy, no systematic attempt has been made to assess hypnotherapy beliefs and use among former trainees in these settings. This study investigated posttraining hypnotherapy use and effectiveness beliefs in a sample of 77 former psychiatry residents and psychology interns. Over 50% of the study sample had sought additional hypnotherapy training beyond the standard lectures and seminars, and almost 30% had attended external hypnotherapy workshops or presentations. Beliefs in hypnotherapy effectiveness were high, but use of hypnotherapy in clinical practice was very low. Former residents and interns who had received supervised training with patients, who had attended hypnosis workshops, and who had a colleague using hypnotherapy were more likely to use hypnotherapy following training. PMID- 8407017 TI - An analysis of hypnotherapist-client sexual intimacy. AB - While sexual interaction between psychologists, physicians, and other health therapists of all kinds and their clients is typically condemned by professional bodies as unethical, the controversy regarding the potential for hypnosis to produce compliant behavior in unwilling or nonconsenting subjects suggests that hypnotherapist-client sex may warrant special attention. Because the experiments required to clarify the potential for hypnosis to potentiate nontrivial compliance are themselves unethical and/or inconclusive, experimental methods cannot be adequately used to clarify this issue. Instead, the matter can be addressed by reference to other forms of evidence, such as the responses of therapists and clients to anonymous surveys and the analysis of cases, that have reached the courts. Consideration of this qualitatively deficient evidence suggests that even if the use of hypnotic suggestion can lead to compliance to sexual demands, overt coercion is seldom used in practice. Social psychological and situational factors are particularly salient in understanding therapist client sex. The question of whether there are special properties of the dynamics of the hypnotic experience, other than specific coercive suggestion and beyond those typically found in other forms of therapy, is considered. Comparisons are drawn with other examples of socially condemned sex, such as teacher-student sex, sexual harassment in the workplace, incest, and extramarital sex. PMID- 8407018 TI - The effects of cue level, hypnotizability, and state instruction on responses to leading questions. AB - Two sessions were conducted in which independent groups of 86 high- and 85 low susceptible subjects, responding individually under waking or hypnotic instruction, answered high- and low-cued leading questions about a video event that depicted shooting at an airport. The two sessions were separated by 1 week, and the same questions were asked in both sessions. It was predicted that highly susceptible subjects responding under hypnotic instruction would show the most evidence of accepting false information via strongly cued leading questions. Results showed general effects for leading questions and level of susceptibility but no firm support for the involvement of hypnosis. Data are discussed in terms of both the linguistic and social factors that appear to have operated on subjects in the study, results overall highlight the strong influence of level of susceptibility on subjects' acceptance of false information. PMID- 8407019 TI - The effects of direct versus indirect hypnotic suggestion on pain in a cold pressor task. AB - Past studies have investigated the usefulness of hypnosis in pain reduction. Although hypnotic analgesia has been found to be effective, it is generally only those subjects who are highly susceptible to hypnosis who benefit. Some experimenters have found that even low-susceptible subjects can use hypnotic analgesia, if the hypnotic induction uses indirect rather than direct hypnotic suggestions. In the present study, high- and low-susceptible subjects were tested for analgesia using either direct or indirect hypnotic suggestion on pain in a cold pressor task. Findings suggest that high susceptibles experience greater pain reduction than do low susceptibles. However, no significant differences were found between the pain reduction in the direct versus the indirect hypnotic suggestion groups. Possible explanations for this lack of differences are discussed. PMID- 8407020 TI - Volatile substance misuse in children and youth: a consideration of theories. AB - A paucity of research exists in the social sciences regarding the theories explaining volatile substance misuse behavior in children and adolescent. Volatile substances include solvents and inhalants such as modeling glue, aerosol sprays, and gasoline. The explanations found in the literature neither sufficiently nor adequately explain possible motivation and causation. Cultural ideas about life and alcohol, as perceived by adult community members, are significant contributing and motivating variables in children's choices to misuse volatile substances. The social forces within cultural frameworks must be acknowledged to better understand youthful choices to misuse substances. PMID- 8407021 TI - A note on waiting lists and demand estimation. AB - Much debate has taken place in the public policy arena concerning the issue of treatment on demand. This article provides a brief discussion of the potential uses of waiting list information in estimating the level of capacity required to address "expressed" demand. A model developed for this purpose is amenable to integration with planning and budgeting processes. Limitations of the model are examined, and possible directions for refinement are suggested. PMID- 8407022 TI - Drinking in the injury event: a comparison of emergency room populations in the United States, Mexico, and Spain. AB - Variables related to drinking in the injury event were compared among probability samples of emergency room patients in Contra Costa County, California (N = 1,001), Mexico City (N = 1,688) and Barcelona, Spain (N = 1,684). Drinking companions and places of drinking prior to injury, place of injury associated with drinking, amount of alcohol consumed, proximity of drinking with the injury event, perceived drunkenness at the time, and causal attribution of drinking with the event were all found to vary among the samples. The data suggest that the context in which alcohol is involved in the injury event is affected by the context in which alcohol is typically consumed in a culture and is important in analyzing alcohol's role in injury occurrence and situations which may be considered high-risk for alcohol-related injuries. PMID- 8407023 TI - Community attitudes toward drug use: the need to assess community norms. AB - Four neighborhoods at high-risk for alcohol and drug problems were surveyed concerning residents' perceptions of the extent and nature of these problems in their neighborhood. Residents were found to have relatively mild perceptions of the extent of misuse but relatively significant experiences with problems caused by drug abuse. They lacked knowledge of treatment and prevention programs. Significant differences were noted among the neighborhoods. Results are discussed in terms of the role of evaluation in community-based programs and the need to survey community norms to plan such programs. PMID- 8407024 TI - Young women and alcohol: psychosocial factors associated with their own drinking, their fathers' drinking, and both. AB - Psychosocial factors associated with young women's own alcohol consumption, and that of their fathers, were assessed in a sample of 231 females (mean age: 18.0 years) from the Hunter Region, NSW, Australia. Consistent with prior research, we found some evidence of poorer adjustment in girls who drank heavily and among daughters of high drinkers. In addition, we found that the "psychosocial profiles" associated with girls' and fathers' drinking were quite different, which argues against a simple additive model which might predict poorer adjustment in high drinking daughters of high drinking men. One interesting finding was that discordance between girls' and fathers' drinking patterns was independently related to some psychosocial problems. PMID- 8407025 TI - Psychosocial predictors of needle sharing among intravenous drug users. AB - This study examined the relationship between health beliefs and needle sharing in a sample of 226 heroin detoxification clients. Zero-order correlations revealed that needle sharing was positively associated with perceived susceptibility and AIDS anxiety, and negatively associated with self-efficacy and communication skill in negotiating safe needle practices. Logistic regression analysis showed that demographic and situational factors also play a role in needle sharing. White race and injection of drugs in shooting galleries or other public places increase the odds of sharing a "dirty" needle, while that probability decreases sharply as a function of drug users' self-efficacy. This last finding suggests that preventive clinical interventions such as counseling or psychoeducational groups maybe needed to help drug users develop confidence in resisting needle sharing. PMID- 8407026 TI - Methadone maintenance and needle/syringe sharing. AB - Drug users who inject drugs while in treatment share needles/syringes less often than users not in treatment. This relationship may reflect treatment processes, such as cognitive or normative change, by which treatment clients are influenced to lower their HIV infection risk. However, reduced needle/syringe sharing among treatment clients may instead be simply a collateral result of reduced injection frequency. In this sample of injection drug users, those who continued to inject while in methadone maintenance treatment reported less sharing than users not in methadone maintenance. This relationship persisted after injection frequency and drug-user background characteristics were controlled. Efforts to identify explanatory treatment processes were, however, not successful. PMID- 8407027 TI - A preliminary ethnographic decision tree model of injection drug users' (IDUs) needle sharing. AB - This article presents a preliminary ethnographic decision tree model of the needle sharing and HIV risk decision-making processes common to injection drug users (IDUs) in Houston, Texas. Ethnographic tree decision modeling is a rigorous qualitative method used to understand and predict how and why people in certain groups do things the way they do. The model was developed from data collected from focus group and individual interviews and naturalistic observations. The research participants were White, African-American and Latino male and female IDUs. The model presents IDUs' needle-sharing routines or scripts based on several prioritized criteria. The model supports other researchers' findings that social roles play an important part in shaping IDUs' needle sharing. Yet, the model attempts to specify the different kinds of roles and relationships. The model suggests that social roles IDUs play and the status hierarchy between roles are the translation mechanisms that organize IDUs' social relationships into drug and HIV risky activities. PMID- 8407028 TI - A longitudinal study of the predictors of the adverse effects of alcohol and marijuana/hashish use among a cohort of high risk youths. AB - Data gathered from a longitudinal study of juvenile detainees in a Southeastern state are used to examine the demographic and life experience factors relating to their reported adverse effects of the use of alcohol and other drugs. The results indicate: 1) marijuana/hashish use, (2) their use of these substances tends to be a vehicle for the expression of personal difficulties in the areas of self-esteem and emotional/psychological functioning, and 3) their use of these two substances needs to be seen in holistic terms. Research and policy implications of the results are drawn. PMID- 8407029 TI - An alternative method for sample size determination in substance misuse prevention research. AB - There is considerable evidence that social science researchers often fail to adequately address statistical power and related sample size issues. This tendency has been particularly salient in substance misuse prevention research. Although failure to carefully address statistical power and sample size issues most frequently results in studies lacking statistical power, other problems can also occur. For example, sample size determination in controlled studies which focus on rates of substance initiation and similar measures require close consideration of baseline rates indicated by these measures. Otherwise, when conventional procedures for sample size determination are utilized, samples in these studies can exceed the size necessary for predesignated levels of power. This article presents a simple alternative to conventional procedures for sample size determination which can be applied to controlled study of substance initiation and similar outcomes. PMID- 8407030 TI - Ethnic identification and alcohol use among Canadian-born and foreign-born high school students in Toronto. AB - Unlike other demographic and sociopsychological correlates of student alcohol and other drug use, the variable of ethnic identification and its possible effects on student substance use have not received adequate attention from researchers. This paper seeks to examine the relationship between ethnic identification and alcohol use on the basis of data collected from a survey of 667 high school students in Toronto. Results show that when other variables (sociodemographic, school, family, availability, and informal social control) are controlled for, the relationship between ethnic identification and alcohol use remains significant in Canadian-born students but not in foreign-born students. Also, foreign-born students are found to exhibit lower levels of alcohol use than their Canadian born counterparts, regardless of cultural origin. Implications of the findings for future studies of ethnicity and alcohol/drug use and for prevention efforts are discussed. PMID- 8407031 TI - Childhood experiences and complicated grief: a study of adult children of alcoholics. AB - For the most part, grief research concentrates on type of loss (e.g., loss of spouse, parent, or child) and/or type of death (e.g., expected or sudden). In contrast, the present paper focuses on a category of persons generally assumed to have had troubled childhoods, adult children of alcoholics (ACOAs). Because of assumed problematic histories, the grief process of ACOAs should be expected to differ from the grief process of non-ACOAs. Using both quantitative and qualitative techniques, 27 ACOAs and 20 non-ACOAs, recruited by newspaper, radio, and word-of-mouth, are compared across characteristics generally associated with ACOAs and/or unresolved grief. Implications for counseling are presented. PMID- 8407032 TI - Perceived client and program moderators of successful therapeutic community treatment for drug addiction. AB - Research examining moderators of therapeutic community treatment outcomes has tended to emphasize the importance of length of stay in programs. To this extent the research literature implies a causal role for treatment in the determination of treatment outcomes. Using the Delphi technique, a panel of residents, staff members, and other drug treatment agency staff involved with the Odyssey House therapeutic community in Melbourne were surveyed regarding their perceptions of factors moderating successful treatment in this program. Examination of the perceptions of those involved with the delivery of therapeutic community treatment suggested a strong respect for the importance of client motivational factors in the determination of treatment outcomes. It is suggested that future research in this area could profitably focus on attempts to quantify the separate contribution of treatment and nontreatment factors in explaining treatment outcomes. PMID- 8407033 TI - Treatment outcome for substance misuse patients with personality disorder. AB - Treatment outcome was compared for three groups of patients in a chemical dependency unit--14 patients with personality disorder, 16 patients with traits of personality disorder, and 34 patients with no personality disorder. Patients with personality disorder were as likely as other patients to complete the 4 month aftercare program and to maintain abstinence while in the aftercare program. PMID- 8407034 TI - Knee function after anterior cruciate ligament ruptures treated conservatively. AB - Thirty-nine patients with ruptures of the anterior cruciate ligament (ACL) were treated conservatively and were subsequently examined at an average of 5.7 years after injury, the uninjured leg acting as a control. The Tegner activity score was significantly lower than the desired activity level. No patients were free of symptoms and only two could take part in sport which involved pivoting. The isokinetic knee extensor and flexor torques, as well as the one-leg-hop and instrumented knee joint laxity tests, were significantly impaired at follow-up. Functional impairment was not related to tests of knee joint laxity. Few patients were pleased with their subjective knee function after an ACL rupture despite thorough initial rehabilitation. PMID- 8407035 TI - In vitro study of contact area and pressure distribution in the human knee after partial and total meniscectomy. AB - Many investigators have attempted to find the cause of the osteoarthritic changes after meniscectomy. Alteration of the mechanical factors resulting in stress concentration, is now thought to be one of the most important causes but few experimental studies have reported the differences in contact area and pressure distribution after partial or total meniscectomy. By using pressure sensitive film, we have calculated the contact area and the pattern of weight distribution in three different situations; intact meniscus, partial and total meniscectomy. The experimental materials were obtained from 5 above knee amputation specimens. The knee joint was fixed in full extension to an Instron machine using an aluminium box and mounting resin. Load was transmitted to the tibiofemoral joint containing the special film, within a physiological range. Analysis of the contact area for each situation (intact meniscus, partial and total meniscectomy) was made by reviewing the film. By measuring the contact area after meniscectomy, we showed that the meniscus performs a load transmitting function in the knee joint. The medial contact area of the tibiofemoral joint with an intact meniscus is always larger than the lateral compartment (1.36:1), but in partial and total meniscectomy the difference between them gradually decreased. There was a minor decrease in contact area after partial meniscectomy and a much greater decrease after total meniscectomy. The degree of stress concentration in the contact area was increased when part or all the meniscus was excised. There was little change of contact area in the opposite, intact side of the joint after partial meniscectomy, but marked change after total meniscectomy. PMID- 8407036 TI - Results of conservative treatment of partial tears of the anterior cruciate ligament. AB - The diagnosis of an acute partial tear of the anterior cruciate ligament was made in 56 patients who did not undergo surgical repair. After a period of up to 5 years, 39 returned for follow up evaluation. They were divided into three groups according to clinical analysis of the knee and the Lysholm-Gillquist score. Our aim was to determine their long term functional limitations. 56% had progressed to anterior cruciate ligament deficiency at the time of follow up. This came about not only after resuming sporting activities, but also occurred in those who were not so active. Our results suggests that a partial tear leaves an irreversible defect which may progress to a complete tear, especially in young athletes engaged in active sport. PMID- 8407037 TI - The MacIntosh lateral substitution reconstruction for anterior cruciate deficiency. AB - Twenty-five patients who had anterior cruciate deficient knees treated by the MacIntosh reconstruction operation were reviewed after 5 years, with particular reference to any changes occurring over time. Subjectively, the knees were improved and the results were even better after 5 years than after one year. This was considered to be due to a decrease in the restriction of flexion, and to the psychological and functional adaptation to the injured knee. In the knees operated on, the anterolateral instability increased significantly during the follow up from one year (mean 1.9 mm) to 5 years (3.0 mm), but the numerical value was still as good as in the intact knees (3.5 mm). PMID- 8407038 TI - Results of arthroscopic acromioplasty related to rotator cuff lesions. AB - Seventy-nine patients with a history of the subacromial impingement syndrome were treated by arthroscopic acromioplasty. The results were evaluated with a new scoring system designed to assess subjective symptoms before and after operation. The outcome was related to the pathological lesions of the rotator cuff seen at arthroscopy and graded by our modification of the Neer classification. The overall results were satisfactory in 67%, with men having better results than women. Those with a history of injury did better than those without, and the results in those with partial or total cuff tears were better than those with intact cuffs. Since the state of the cuff affects the result, arthroscopy should precede acromioplasty. PMID- 8407039 TI - Musculocutaneous nerve entrapment in the upper arm. AB - Wasting and weakness of the biceps and brachialis muscles can occur when the musculocutaneous nerve is compressed as it passes through the coracobrachialis muscle; there may also be impairment of sensation on the lateral aspect of the forearm. In our patient, symptoms appeared after strenuous exercise which included more than 500 press-ups each day. Electromyography and telethermic examination confirmed the diagnosis. He was advised to stop his strenuous exercises and within 3 months muscle strength and sensation had returned. PMID- 8407040 TI - Congenital absence of the radius. AB - Between 1971 and 1987, we treated 26 patients with congenital absence of the radius which had produced a club hand; 37 hands were affected. Treatment before the age of 8 months was by gentle manipulation and serial plasters. If a functional position was not obtained, and in patients over 8 months, surgical reduction was carried out. A modified Riordan procedure was used until 1983, and thereafter a double Z-plasty. The best results were achieved in patients under 4 years of age. PMID- 8407041 TI - Hemovac drainage after hip arthroplasty. AB - The authors have reviewed the pattern of drainage of Hemovac devices in 107 arthroplasties of the hip. Drainage ceased in 55 hips (51%) within 18 hours and 93 hips (89%) within 24 hours. The mean drainage volume was 303 cc in femoral head replacement, 498 cc in simple total hip replacement, and 663 cc in extensive total hip replacement. Ninety nine per cent of the total drainage had been achieved within 24 hours, indicating that drainage devices can be removed then rather than between 24 and 72 hours as had been previously recommended. This early removal will help reduce the possibility of retrograde infection, and promote more comfortable rehabilitation after operation. PMID- 8407042 TI - Magnetic resonance imaging in the evaluation of the lumbar herniated intervertebral disc. AB - Two hundred and eleven patients with lumbar disc herniation at 242 levels were divided into 5 groups by their appearances on magnetic resonance imaging (MRI), and the findings at operation were compared to assess the accuracy of the MRI classification. There were no negative explorations. There was 92% sensitivity, 91% specificity and 92% accuracy for MRI in distinguishing protruded discs from other forms of lumbar disc herniation. For sequestrated discs there was 92% sensitivity, 99% specificity and 97% accuracy. In the extruded subligamentous type there was 71% sensitivity, 82% specificity and 79% accuracy, and 52% sensitivity, 92% specificity and 81% accuracy in the extruded transligamentous type. The overall accuracy of MRI predicting the types of herniated lumbar intervertebral disc was 85%. High resolution MRI is sensitive in detecting disc disease and specific in characterizing various subgroups of disc herniation, especially those which are sequestrated. PMID- 8407043 TI - Magnetic resonance imaging (MRI) of a benign cystic teratoma in the gluteus region. AB - Magnetic resonance imaging (MRI) was used to help in the diagnosis of a teratoma in the gluteus maximus region. As far as we know no case of this kind of tumour has been investigated with MRI examination. PMID- 8407044 TI - Irregularity of the apophysis of the ischial tuberosity. AB - We report five cases of irregularity of the apophysis of the ischial tuberosity (Kremser's disease). In four, plain radiographs showed that the density of the radiolucent area increased within several months of a minor injury. Serial computerised tomography demonstrated new bone formation across the widened apophyseal plate, although there was no change in the shape or density of the apophysis itself. The separation repaired and remodelled spontaneously within a year. In one case, CT showed that the apophyseal ossification centre was smaller than that on the opposite side. Our findings showed that in four cases the irregularity of the apophysis of the ischial tuberosity may have been caused by an avulsion fracture, but the appearance in the fifth case was a normal variant of the ossification centre. PMID- 8407045 TI - Arthrodiastasis for stiff hips in young patients. AB - Joint distraction (arthrodiastasis) with a unilateral fixator was used to treat 9 patients with stiffness of the hip which had followed Perthes' disease (3), epiphysiolysis (2), congenital dysplasia (2), tuberculosis (1) and idiopathic chondrolysis (1). Their average age was 14 years, and they all had pain, limp and shortening of the leg. Distraction of 0.5 to 1 cm was maintained for an average of 94 days. The average range of movement subsequently was 65 degrees compared with 20 degrees before. The articular space was widened by an average of 2.8 mm, and only 3 patients had pain on follow up. PMID- 8407046 TI - Bone reaction to uncemented threaded polyethylene acetabular components. AB - One hundred and eighty-one cementless total hip arthroplasties with threaded polyethylene acetabular components, carried out between 1980 and 1981, have been reviewed retrospectively. Discrepancy between the complaints of the patients with loose components and destruction of acetabular bone led to the development of a four stage radiological grading system which would enable loosening to be anticipated in routine cases. There was a high percentage of loosening (33%) in our series and direct bone-polyethylene contact gave rise to severe granulomatous reactions around the implants. Strict radiological follow up and early revision surgery is indicated in this type of arthroplasty. PMID- 8407047 TI - Trabecular patterns of the calcaneum as an index of osteoporosis. A comparison using gamma-absorptiometry. AB - Measurements of bone mineral content in the calcaneum were made by gamma absorptiometry in 77 patients with ankle fractures treated by operation, and compared with the calcaneal osteoporosis index. The index was calculated from plain lateral radiographs of the calcaneum. The bone mineral measurements showed a wide range, but there was only a narrow range of the index with exclusively high values. There was a weak correlation between the calcaneal index and the bone mineral content in the injured ankles, but no or only a very poor correlation in the uninjured ankles. We also found no correlation between the decline in index and loss of mineral content in the injured ankles. PMID- 8407048 TI - Renal osteodystrophy in children: correlation between aetiology of the renal disease and the frequency of bone and articular lesions. AB - In a retrospective study of 70 patients with chronic renal failure, 66% were found to have manifestations of renal osteodystrophy. Children with either hereditary or congenital renal diseases were more prone to develop bone lesions than patients with acquired renal diseases. PMID- 8407049 TI - Impaired cytosolic free calcium response in splenic T-cells from mice fed with ethanol-containing diet. AB - Calcium-dependent signal transduction pathways of T-cell proliferation have been extensively studied in the past years. However, little is known about effects of ethanol on the calcium-dependent signal transduction pathway in T-cell proliferation. Thus, a murine model was used to determine effects of ethanol in vivo on T-cell proliferation and the intracellular free calcium concentration [Ca2+]i in response to Concanavalin A (Con A) and recombinant IL2 (rIL2) in T cells. Splenic cells from young C57BL/6 mice, that had been fed on 3 different diets (ethanol-, maltose substitute- and standard liquid-diet) for 7-8 weeks were tested for their proliferative responses to Con A and rIL2. Concurrently, measurement was also made of [Ca2+]i in the nylon-wool-enriched resting T-cells induced by Con A and in Con-A-activated blast T-cells induced by rIL2. Our results showed that [Ca2+]i increases were seen in the splenic T-cells from three different groups of mice following Con A, but not rIL2 stimulation. However, this increase was much smaller in the splenic T-cells from ethanol-fed mice as compared to mice on maltose- or standard-diet. Furthermore, we also demonstrated that the impaired [Ca2+]i increase was seen in the T-cells of the same ethanol fed mice having decreased the proliferative response to Con A. This reduced proliferation did not result from the presence of excessive suppressor T-cell activity. Finally, we also demonstrated that both the number of IL2 binding sites/cell and the Kd values of the low- and high-affinity IL2R on the T-cells from ethanol-fed mice were unaltered. Because evidence indicates that (1) a normal level of [Ca2+]i increase is a prerequisite for the production of IL2 by mitogen-stimulated T-cells, and (2) T-cells from ethanol-fed mice have normal capacities to produce IL2 that is the crucial growth factor controlling T-cells to progress through the cell cycle, these lines of evidence taken together with the results of this study suggest that the impairment in [Ca2+]i increases in T cells from ethanol-fed mice may not be the primary factor contributing to the diminished T-cell proliferation in the same mice. PMID- 8407050 TI - Non-specific resistance induced by a low-toxic lipid A analogue, DT-5461, in murine salmonellosis. AB - The ability of DT-5461, a chemically synthetic low-toxic lipid A analogue, to activate anti-Salmonella activity in C3H/HeN mice was examined. Previous intraperitoneal (i.p.) injection of DT-5461 (100 micrograms or more/mouse) significantly hindered the bacterial growth in the peritoneal cavities of the mice after the i.p. infection with Salmonella typhimurium LT2 strain. The effect was the maximum when DT-5461 was given 6 h before the challenge. The injection of DT-5461 6 h in advance could also confer protection against the infection. Bactericidal activity enhancement was also seen in mice that had been injected with a small amount of recombinant murine IFN-gamma (10(3) U per mouse) and non effective dose (10 micrograms) of DT-5461 together 6 h before the challenge. Bactericidal effect enhancement was seen in mice that had been injected with IFN gamma at 6 h and DT-5461 at 3 h before the challenge, while it could be hardly seen in mice injected with them in a reversed order. The i.p. injection of DT 5461 recruits the exudate cells into the peritoneal cavities, and the phagocytic and bactericidal abilities of the macrophages in the exudate cells are apparently elevated. The mechanisms of non-specific resistance enhancement induced by DT 5461 were discussed. PMID- 8407051 TI - Delta-9-tetrahydrocannabinol-(THC)-mediated inhibition of macrophage macromolecular metabolism is antagonized by human serum proteins and by cell surface proteins. AB - Our previous study demonstrated THC-inhibited DNA synthesis and the phagocytic activity of P388D1 cells [Tang, Lancz, Specter & Bullock (1992) Int. J. Immunopharmac., 14, 253-262]. The ability of proteins in human and bovine sera and of constitutive cellular proteins to modulate the biologic activity of THC was investigated. Both human and fetal bovine sera antagonized a THC-mediated inhibition of P388D1 cell DNA synthesis in a dose-dependent manner. This antagonism was proportional to the protein concentration present in the medium. Both albumin and gamma-globulins influenced THC's inhibitory effects, although they were less potent alpha/beta serum lipoproteins. Exclusion of fatty acid moieties from the albumin did not diminish its ability to antagonize THC. Tritium labeled THC was acid precipitable only after incubation with bovine or human serum albumin but not DNA, suggesting a physical interaction between the cannabinoid and the protein. Further studies showed that pre-treating cells with trypsin to remove surface proteins significantly enhanced the inhibitory activity of sub-toxic concentrations of THC. Thus, the data indicate that the magnitude of THC's biological effects is determined by the presence and concentration of soluble proteins in the microenvironment and by constitutive proteins present on the cell surface. PMID- 8407052 TI - Sustained zidovudine treatment on hematopoiesis in immunodeficient mice. AB - Zidovudine (AZT) has been the drug of choice in the treatment of human AIDS; however, associated with the use of zidovudine has been the development of hematopoietic toxicity, the mechanism of which is not clearly defined. We report here studies designed to evaluate dose-escalation of zidovudine, i.e. 0.1 and 1.0 mg/ml placed in the drinking water on hematopoiesis in C57BL/6 normal and LP-BM5 immunodeficiency virus-infected mice. Over a 6-week evaluation period, compared to normal, non-virus-infected controls, murine immunodeficiency (MAIDS) infection was associated with reduced hematopoietic progenitors, i.e. CFU-E, BFU-E, CFU-GM, and CFU-Meg from bone marrow and spleen. Following zidovudine treatment, further suppression of marrow-derived progenitors was observed, while increased numbers of progenitors were obtained from the spleen. Spleen-derived erythroid progenitors, i.e. CFU-E, were increased by 950% (P < 0.001) from MAIDS-infected animals receiving 1.0 mg/ml of drug following 4-weeks exposure compared to non drug-treated MAIDS control animals. Splenic BFU-E were increased 654% following 6 weeks exposure compared to non-drug-treated MAIDS-infected mice. This study suggests that the bone marrow is particularly sensitive to zidovudine toxicity which, at least early in exposure, appears to be compensated by splenic-derived hematopoiesis, in particular, erythropoiesis. Overt toxicity develops when, at least in this immunodeficiency model, the spleen is unable to provide progenitors in response to continued zidovudine exposure in vivo. PMID- 8407053 TI - The pectic polysaccharide from Bupleurum falcatum L. enhances immune-complexes binding to peritoneal macrophages through Fc receptor expression. AB - Binding of glucose oxidase-anti-glucose oxidase complexes (GAG), a model of immune complexes, to macrophages was enhanced by treatment with an acidic pectic polysaccharide, bupleuran 2IIb, from Bupleurum falcatum L. GAG binding to macrophages by bupleuran 2IIb increased in a dose-dependent fashion, and was abolished when the Pronase-treated macrophages were incubated with bupleuran 2IIb. The GAG binding enhancing activity of bupleuran 2IIb was reduced by periodate oxidation but not Pronase digestion of bupleuran 2IIb. When bupleuran 2IIb was digested with endo-polygalacturonase, the resulting enzyme resistant carbohydrate portion showed potent activity. Scatchard analysis indicated enhanced expression of the Fc receptor (FcR) on the surface by the action of bupleuran 2IIb. The enhancement of GAG binding by bupleuran 2IIb was inhibited by the presence of actinomycin D or cycloheximide. Bupleuran-2IIb-stimulated cells showed enhanced expression of both FcRI and FcRII mRNA, which were measured as PCR products. These results suggested that the endo-polygalacturonase resistant carbohydrate portion of bupleuran 2IIb is important for the expression of the activity, and that the activity of bupleuran 2IIb on GAG binding was mediated by receptors for polysaccharide on the cells. The up-regulation of the Fc receptor by bupleuran 2IIb was also suggested to mediate by de novo synthesis of the receptor protein. PMID- 8407054 TI - Macrophage functions in drugs of abuse-treated mice. AB - The in vivo effect of morphine, methadone and cocaine on murine peritoneal macrophage-mediated cytostasis and macrophage supernatant mediated cytostasis was investigated. In addition, the activity of drugs of abuse was studied on IL-1 alpha and TNF production by activated murine macrophages. A highly depressive effect on macrophage-mediated cytostatic activity and a decrease of IL-1 alpha and TNF levels were found in the supernatants of activated macrophages observed in morphine- and cocaine-treated mice. Conversely, a significant impairment of macrophage functions was not observed in methadone-treated mice. Our results suggest that the inhibition of macrophage defensive functions caused by drugs of abuse may be an important cofactor in the pathogenesis of infectious diseases. PMID- 8407055 TI - Cocaine reduces macrophage killing by inhibiting reactive nitrogen intermediates. AB - The present study describes the inhibition of macrophage-mediated cytotoxicity (MMC) by cocaine and suggests a possible mechanism. Mice (C57BL/6) were injected i.p. with cocaine. At various intervals after exposure to cocaine, peritoneal macrophages (M phi) were removed, cultured in the presence of interferon gamma and LPS, then incubated with 51Cr labeled target cells. A single injection of > or = 10 mg/kg cocaine was sufficient to inhibit cytotoxicity to P815 cells. This inhibition was evident 3 h after exposure to cocaine and could still be demonstrated 24 h later. Since reactive nitrogen intermediates (RNI) have been reported to be one of the major mechanisms by which M phi kill, the amounts of NO2- produced by M phi from cocaine-injected animals were compared with that produced by equivalent controls. Cocaine reduced the level of NO2- in a dose dependent manner which correlated with MMC. There was a significant reduction in NO2- produced by activated M phi, 3 h after i.p. injection of cocaine but not at 24 h, using > or = 5 mg/kg. At 12 h there were differences between M phi from control animals and animals receiving > or = 10 mg/kg cocaine. By 24 h there were no differences between control and cocaine-injected animals even at the highest dose employed (25 mg/kg). These results suggest that cocaine reduces the killing ability of murine M phi through a temporary reduction of RNI. PMID- 8407056 TI - Enhancement of T-cell proliferation by PSK. AB - Protein-bound polysaccharide, PSK, is a biological response modifier and influences various immunological functions in vivo, including those of T-cells. However, PSK has not been proved to stimulate proliferation of T-cells in vitro in contrast to various lines of evidence that indicate the proliferation of B cells in vitro. PSK enhanced the proliferation of spleen cells in C3H/He and nude mice in vitro. In this study such proliferating cells were detected by monoclonal antibody to bromodeoxyuridine (BrdU) incorporated into DNA, and their subsets were determined by flowcytometry with monoclonal antibody to Thy1 as a cell surface marker of T-cells. When spleen cells from C3H/He mice were cultured with PSK for 3 days, the percentage of BrdU positive cells increased, and about 67% of BrdU positive cells were Thy1.2 positive. In addition, PSK also had the activity to enhance the proliferation in a T-cell-enriched fraction from spleen cells by nylon fiber columns as well as that in original spleen cells. These results suggest that PSK enhances the proliferation of T-cells as well as B-cells. PMID- 8407057 TI - Restorative effect of short term administration of thymulin on thymus-dependent antibody production in restraint-stressed mice. AB - Thymulin is a well defined nonapeptide produced by thymic epithelial cells, and plays an important role in thymocyte differentiation. We investigated the restorative effect of thymulin on the stress-induced reduction in the production of antibodies against SRBC in mice. Antibody production in stressed mice was reduced to about 50% of that in normal mice. A 3-day period of administration of thymulin (1 ng/kg) restored antibody production to close to the normal level, whereas thymulin (1 ng-1 microgram/kg) did not affect antibody production in normal mice. These results indicated that thymulin normalizes the altered thymus dependent immune responses. PMID- 8407058 TI - Tissue specificity of translation inhibition in Sprague-Dawley rats following in vivo cyclosporin A. AB - We have shown that following the in vivo administration of cyclosporin A (CsA) to either male or female Sprague-Dawley rats, there is a time- and dose-dependent inhibition of translation elongation in renal microsomes. Experiments reported in this paper explore the tissue specificity of translation inhibition. Following 6 days of oral 50 mg/kg/day CsA or control vehicle, animals were sacrificed and renal, hepatic and cardiac microsomal and cytoplasmic fractions prepared for "run off" translation assays. These assays demonstrated that in vivo CsA resulted in a reduction in renal microsomal 3H-L-leucine incorporation to 25% of control values, a reduction of cardiac microsomal incorporation to 60% of controls and a stimulation of hepatic microsomal incorporation to 140% of controls. Cross-over experiments involving the addition of renal cytoplasmic fractions from CsA treated animals to renal microsomal fractions from control-vehicle-treated animals depressed 3H-L-leucine incorporation to approximately 50% of control values. When the renal cytoplasmic fraction from CsA-treated animals was added to renal, hepatic or cardiac microsomal fractions from control-vehicle-treated animals, 3H-L-leucine incorporation values were consistently reduced to approximately 50% of controls. We have reported that translation elongation is inhibited by renal cytoplasm from CsA-treated rats in the presence of control renal cytoplasm. These data suggest that inhibition arises from the presence of an inhibitor rather than from a deficiency in elongation factors. The data reported in this paper demonstrate that microsomal fractions from various tissues have equal sensitivity to CsA-mediated renal cytoplasmic translation inhibition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407059 TI - Investigation of circadian rhythms in peak power and mean power of female physical education students. AB - The aim of this investigation was to establish if there were characteristic circadian rhythms in peak power and mean power. Thirteen female physical education undergraduate students, aged from 18-21 years, were measured for peak power and mean power (Wingate Anaerobic Test) at four time points: 0300, 0900, 1500, and 2100 hours. Peak power was defined as the highest mechanical power that is elicited during a 5-sec period and mean power was defined as the average power that is sustained throughout a 30-s period. A significant circadian rhythm was found for the peak power (F = 3.22, p = 0.030). The mean peak power at 1500 h was about 7% higher (p < .05) than at 0300 h. Also a tendency to circadian rhythm was observed in mean power (F = 3.99, p = 0.01) being about 16% and 15% higher respectively (p < 0.05) at 1500 h and 2100 h than at 0300 h. Based upon the particular protocol used, it was concluded that there are characteristic circadian rhythms in peak power and mean power. PMID- 8407060 TI - In vivo examination of the dynamic properties of the human heel pad. AB - The shock-absorbing characteristics of the heel pad in vivo were examined in adults (N = 16) and 7-year-old children (N = 5) using a drop-impact tester (wt = 5 kg). Impact velocities were 0.72 m/s and 0.93 m/s. It was found that in adults the average peak deceleration was 11.6 G at an impact velocity of 0.93 m/s. The maximum deformation of the heel pad was 11.3 mm, and the computed energy absorption during impact amounted to 79% (range = 75%-89%). These mechanical characteristics remained nearly the same even after 6 min of repeated impacts by the impact tester and even after a 10 km run. The children had larger values of peak deceleration and maximum deformation and smaller energy loss than the adults. It was concluded that the heel pad was a fairly effective shock attenuator and high energy absorber, and that these characteristics remain nearly unchanged even after a relatively long period of repeated impacts. It was also concluded that the mechanical properties of the children's heel pads were different from those of the adults. PMID- 8407061 TI - Effect of opiate and beta-adrenergic blockers on the gut transit response to mild exercise. AB - Acute mild exercise accelerates transit of material through the gastrointestinal tract via mechanisms that remain entirely unknown. Because exercise increases peripheral sympathetic drive, and can increase circulating opioids, in this study we examined the role that naloxone and propranolol-dependent pathways play in accelerating orocecal transit in exercise. Transit was determined in 11 subjects by H2 detection after ingestion of 0.36 g/kg lactulose in a 240 kcal, 250 ml liquid meal; the exercise was mild walking at 5.6 km/hr. Under naloxone treatment (0.4 mg intravenous bolus followed by infusion at 40 micrograms/kg/hr) transit acceleration in mild exercise was apparent: transit time (mean +/- SE) was 87 +/- 7 min at rest, 63 +/- 5 min in exercise (p < 0.05), a result identical to that seen in previous studies not involving drugs. In contrast, propranolol treatment (160 mg/day) accelerated the resting transit rate to equal the rate during exercise (65 +/- 6 vs. 60 +/- 4 min; p = NS). Under both conditions, exercise elevated oxygen consumption and heart rate, although propranolol predictably produced relative bradycardia at rest and exercise. These results suggest that mild exercise accelerates upper gut transit via an opiate-independent effect that, for gut propulsive motility, yields results similar to beta-adrenergic blockade. PMID- 8407062 TI - Effects of anabolic steroids and endurance exercise on cardiac performance. AB - This study examined the effects of anabolic steroids and exhaustive endurance exercise, and the combination of both treatments on in vitro cardiac contractile function. Fifty-two male Sprague-Dawley rats were randomly assigned, in groups of 13, to one of the four groups: sedentary control (C), steroid-treated (ST), exercise-trained (E), and exercise plus steroid treated (E+S). Nandrolone decanoate was administered to the steroid-treated groups every 7-9 days during the 10 weeks study, while the C and E groups received glycerol injection on these occasions. The exercised rats ran on a treadmill wearing a collar weight (2-3% of body weight) for 50 min, 5 days a week. In vitro ventricular performance was assessed in isolated Langendorff perfused hearts in response to increasing left ventricular balloon volumes. Left ventricular +dP/dt, dP/dt at 60 mmHg developed pressure, tension time index, and Emax (slope of the pressure-volume curves) were significantly (p < 0.05) lowered in the E+S group, as compared to the C, E, or ST group. Meanwhile, the E group exhibited a higher (p < 0.05) left ventricular systolic pressure (LVSP) than the C and E+S groups. No significant differences in dP/dt/P, heart weight (wt), and heart wt to body wt ratio were detected among the four groups. Rats subjected to endurance exercise or anabolic steroid treatment alone exhibited no change in LV function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407063 TI - Serotonergic agonists and antagonists affect endurance performance in the rat. AB - The purpose of this experiment was to examine the effects of administration of serotonergic (5-HT) agonist and antagonist drugs on run-time to exhaustion (RUN EXH) in male and female rats. RUN-EXH was reduced (p < 0.05) in a dose related manner by increasing dosages of quipazine dimaleate (QD: general 5-HT agonist) (0 5 mg.kg-1 i.p.) administered immediately prior to exercise (treadmill running at 20 m.min-1 and 5% grade). Conversely, RUN-EXH was increased (p < 0.05) by the greatest dosage of LY 53,857 (LY: 5-HT1C and 5-HT2 antagonist) (1.5 mg.kg-1 i.p.). Drug effects were similar in male and female rats. The negative effects of QD administration on RUN-EXH were not attenuated by administration of the peripherally restricted antagonist, xylamidine tosylate (up to 200 ug.kg-1 i.p.). The results of this investigation indicated that fatigue during prolonged exercise can be influenced by direct pharmacological administration of a serotonergic agonist and antagonist and that the mechanisms underlying these effects are likely to be central (brain) in nature. PMID- 8407064 TI - Aerobic and anaerobic power characteristics of competitive cyclists in the United States Cycling Federation. AB - The purpose of this study was to characterize the aerobic and anaerobic capabilities of United States Cycling Federation cyclists in different categories. To determine aerobic and anaerobic power, 38 competitive road cyclists (32 males, 6 females) performed a VO2max test and a Wingate anaerobic test, respectively. Male cyclists in category II had the highest VO2max, both in absolute and relative terms. Their VO2max was 6% and 10% higher than category III and IV cyclists, respectively (4.98 +/- 0.14 vs 4.72 +/- 0.15 vs 4.54 +/- 0.12 l/min). A significant difference existed between category II and IV male cyclists (p < 0.05). VO2max for female cyclists (3.37 +/- 0.13 l/min) was significantly (p < 0.05) lower than those for males. The Wingate anaerobic test revealed that male cyclists in category II also had the highest anaerobic power output. The peak power output in category II, III and IV was 13.86 +/- 0.23, 13.55 +/- 0.25, and 12.80 +/- 0.41 W/kg, respectively. The mean power output in category II, III, and IV was 11.22 +/- 0.18, 11.06 +/- 0.15, and 10.40 +/- 0.30 W/kg, respectively. The difference in the mean power output between category II and IV was significant (p < 0.05). Female cyclists recorded significantly less peak and mean power output than their male counterparts (p < 0.05). However, when expressed relative to lean body mass, anaerobic power was similar for both sexes. No inter-correlation was found in any measurement between the aerobic and anaerobic power values. On the whole, category II male cyclists were characterized by higher aerobic and anaerobic power outputs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407065 TI - The use of heart rates to monitor exercise intensity in relation to metabolic variables. AB - Exercise intensity during training and competition was assessed in women runners in relation to two metabolic markers determined in the laboratory, ventilatory threshold (VT) and 4 mM of blood lactic acid (OBLA). Heart rates (HRs) were then obtained during 6 days of training and during an 8 km race. These HRs were used as references for quantifying the amount of hard (> HR-OBLA), moderate (< HR OBLA, > HR-VT) and easy (< HR-VT) intensity training. Mean maximal heart rate (HRmax) was 183 bpm, HR-OBLA was 94.8% of HRmax, and HR-VT was 82.2% of HRmax. Average weekly training time was 301.2 min, of which 45.8% was in the easy intensity range, 45.7% in the moderate intensity range, and 8.9% in the hard intensity range. Self-reports of how training time was spent differed from actual training as revealed by the monitored heart rates. Subjects reported that they completed 3 sessions per week of easy intensity training, less than 1 session of moderate intensity training, and 1.5 sessions of hard intensity training. More than 70% of the 8 km race was performed at an intensity higher than HR-OBLA (173 bpm). Although 8 km race intensity closely corresponded with that at OBLA, very little training time was spent at that running intensity. HR monitoring of training intensity provided more accurate information than self-reports of training intensity. PMID- 8407066 TI - A rare complication of a central venous catheter system (Port-a-Cath). A case report of a catheter embolization after catheter fracture during power training. AB - A 36 year old patient received an implantable central venous catheter system for a bone marrow transplantation. One year after the successful transplantation, embolization of the catheter was discovered by a routine x-ray three weeks after beginning of an exercise program with a spring expander including arm exercises. The catheter was removed without further complication via the vena femoralis. We assume that the cause for this incident was material fatigue due to pressure between clavicula and first rib possibly caused by strength training. We suggest as a consequence that patients with an implanted catheter system should before starting exercise consult a sports medicine specialist who would in turn cooperate with the specialist responsible for the catheter, so that an adequate and safe training program can be selected. PMID- 8407067 TI - Exercise-induced stress injuries to the femur. AB - Seventy-one athletes with 74 stress injuries to the femur were studied using a case-controlled design. Forty-three were females (26.6 yrs) and 28 were males (31.2 yrs). Each patient had exercise-induced pain in the hip, groin or thigh and a Tec-99m-MDP bone scan showing focal uptake of radionuclide in the femur. Running was the most common activity at the time of injury (89.2%) followed by triathlon (4.6%) and aerobic dance (4.6%). Thirty per cent of the runners had increased their training duration immediately prior to their first symptom. Anterior thigh pain was the most frequent site of exercise-induced pain (45.9%) followed by hip pain (27%) and groin pain (8.1%). During the clinical examination, when asked to hop on the affected limb, 70.3% of the patients had pain reproduced in the hip, groin or anterior thigh. There were 39 cases (53%) involving focal uptake of radionuclide in the femoral shaft, 15 (20%) in the lesser trochanter, 11 (15%) in the intertrochanteric region between the femoral neck and the greater trochanter, 8 (11%) in the femoral neck and 1 (1%) in the greater trochanter. Two patients suffered displaced fractures, one at the femoral neck and the other in the shaft of the femur. Neither patient had previously sought medical attention for their leg pain. Of 46 plain radiographs taken, only 11 (24%) were abnormal. The mean time to diagnosis and recovery were 6.6 and 10.4 weeks respectively. Substitution of cycling and water exercise for running were the most common therapeutic interventions. PMID- 8407068 TI - Contact dermatitis. PMID- 8407069 TI - Inherited epidermolysis bullosa. PMID- 8407070 TI - 50% resorcinol peel. PMID- 8407071 TI - Atopic dermatitis: nomenclature. PMID- 8407072 TI - Observations: the dermatologist and Goya. PMID- 8407073 TI - Epidermolysis bullosa in the eastern province of Saudi Arabia. AB - BACKGROUND: Epidermolysis bullosa is recognized to be rare, but its prevalence in the Eastern Province of Saudi Arabia had not been previously established. METHODS: We reviewed 49,902 dermatology cases seen in our clinic over a 7-year period (1984 through 1990) and carried out a therapeutic trial of oral phenytoin in three severe cases of epidermolysis bullosa dystrophica (Recessive dystrophic type). RESULTS: Sixteen cases of epidermolysis bullosa were found in this series, ten (62.5%) of which were of the dermolytic type (epidermolysis bullosa dystrophica), and four (25%) of the epidermolytic type (epidermolysis bullosa simplex). Parental consanguinity was established in fourteen (87.5%) of these cases. All ten cases of epidermolysis bullosa dystrophica developed complications. Three severe cases were treated with oral phenytoin and managed with meticulous nursing and nutrition with blended foods and protein and vitamin supplements and responded satisfactorily. CONCLUSIONS: This study confirmed the rarity of epidermolysis bullosa in this province (population 3,000,000), and demonstrated the usefulness of oral phenytoin therapy, meticulous nursing, and good nutrition in the management of epidermolysis bullosa dystrophica. PMID- 8407074 TI - Update on nutrition and psoriasis. AB - BACKGROUND: It has been established that severe psoriasis may lead to nutrient depletion, especially of protein, folate, and iron. Nutrient loss occurs due to the accelerated loss of nutrients from the hyperproliferation and desquamation of the epidermal layer of skin in psoriasis. We have proposed that nutritional support as a secondary form of therapy may be beneficial in aiding some psoriasis patients return to a state of remission. METHODS: To determine how frequently nutritional abnormalities occur in hospitalized psoriasis patients, a retrospective analysis was done of the nutritional status of 50 patients admitted for the treatment of psoriasis. Chart records were analyzed and laboratory data were interpreted in consideration of concurrent medical problems. Protein status and anemia were the primary nutritional indicators studied. RESULTS: We found that of those parameters that may be used as indicators of nutritional status, 18% of the patients had a decreased total protein, 16% of the patients had a decreased serum albumin, 38% had an elevated mean corpuscular volume, and 39% had a decreased hematocrit. CONCLUSIONS: Our findings support the concept that wide spread psoriasis places the patient at risk to develop minor nutritional abnormalities in protein and folate status even when accounting for confounding medications and coexistent diseases. PMID- 8407075 TI - Psychosocial consequences of rejection and stigma feelings in psoriasis patients. AB - BACKGROUND: Many studies as well as clinical experience indicate that patients often feel stigmatized by their psoriasis and may experience rejection due to their skin disease. METHODS: In this study of connections between feelings about having psoriasis, rejection by others, and the impact on work experience, alcohol use, and psychiatric help-seeking, 100 adults with moderate to severe psoriasis responded to a specially designed questionnaire. Multiple regression analysis is used to determine whether, and how, rejection experience and stigma feelings are associated with the three psychosocial outcomes after controlling for psoriasis severity. RESULTS: Nineteen percent of the subjects experienced 50 episodes of gross rejection as a result of their psoriasis, most often from a gym, pool, hairdresser, or job. Rejection experience leads to feeling stigmatized, which is then associated with help seeking and, to some degree, with interference with work. Rejection can lead to feeling stigmatized and to increased alcohol consumption, but the data show that patients may drink without conscious awareness of their feelings. CONCLUSIONS: When patients experience psoriasis related rejection, they may feel stigmatized and suffer further adverse effects on their emotional and occupational life. It is urgent that ways of helping people deal with the impact of psoriasis, its stigmatizing potential, and the consequences of psoriasis-induced rejection be emphasized. PMID- 8407076 TI - Epidermolysis bullosa simplex associated with spinal muscle atrophy. PMID- 8407077 TI - An uncommon presentation of multiple symmetric lipomatosis. PMID- 8407078 TI - Psoriasis contralateral to hemiparesis following cerebrovascular accident. PMID- 8407079 TI - Acral persistent papular mucinosis. PMID- 8407080 TI - Mal de Meleda in a Laotian family. PMID- 8407081 TI - Yellow nail syndrome resolution following treatment of pulmonary tuberculosis. PMID- 8407082 TI - Cholesterol emboli syndrome. PMID- 8407083 TI - Intralesional treatment of cutaneous leishmaniasis with sodium stibogluconate antimony. AB - BACKGROUND: Cutaneous leishmaniasis represents a difficult disease to manage in endemic areas. Systemic treatment is hampered by both expense and compliance. Side effects may play a major role in this aspect as well. METHODS: The effectiveness of intralesional treatment of leishmaniasis was investigated. Seven hundred and ten patients were treated with injections of sodium stibogluconate intralesionally. The clinical diagnosis was confirmed by demonstrating the parasite in the smears obtained from the lesion. Fine insulin needle was used to infiltrate the lesion with sodium stibogluconate (0.5 to 1.0 mL). RESULTS: Generally eight injections were sufficient, but some of the complicated lesions needed up to 24 injections. Sixty-two percent of patients were men. The majority of the study population (64%) were children below 15 years of age. The results showed that 72% of lesions healed completely, 23.9% showed some improvement, while 4.1% showed some deterioration. Lesions of the lips, cheeks, chin, and neck healed faster than lesions in other parts of the body. Side effects were mild and limited to pain at the site of the injection and hyperpigmentation in those who were treated by folk medicine. CONCLUSIONS: Intralesional treatment is as effective as the standard systemic antimonials. It offers a less expensive alternative and a low side effects profile. Our findings confirmed the findings of earlier workers. It is recommended for treatment of cutaneous leishmaniasis in endemic areas. PMID- 8407084 TI - History of dermatology and venereology in Singapore. PMID- 8407085 TI - Anticardiolipin antibodies and lower leg plaques and ulcers of 45 years duration. PMID- 8407086 TI - Cutaneous hypersensitivity reactions in patients with AIDS. PMID- 8407087 TI - Atopic dermatitis: clinical criteria. PMID- 8407088 TI - Treatment of dermatophytoses II: Newer topical antifungal drugs. PMID- 8407089 TI - Tropical dermatology. PMID- 8407090 TI - Classification of lupus erythematosus. PMID- 8407091 TI - Preliminary, dermatologic first step criteria for lupus erythematosus and second step criteria for systemic lupus erythematosus. AB - BACKGROUND: Comparisons of cases of systemic lupus erythematosus (SLE) with cases of rheumatoid arthritis and other rheumatologic disorders affords the basis of the 1982 revised criteria of the American Rheumatism Association (ARA) for classifying SLE cases. We address three questions: Do comparisons of LE cases with non-LE cases that have suggestive skin lesions yield criteria for use in dermatology clinics for primary classification of cases with photo distributions of skin lesions? Do comparisons of SLE with cutaneous LE cases yield the same or similar criteria to the revised ARA criteria for SLE? How should subacute cutaneous LE cases be evaluated for signs of significant systemic involvement? METHODS: Discriminant analyses on 168 cases with skin lesions suggestive of LE were performed using data based on the ARA criteria for SLE and study factors for cutaneous LE suggested by the European Academy of Dermatology and Venereology. RESULTS: These yielded two sets of criteria: (1) The 11 preliminary, dermatologic first step criteria (10 plus 1 for discoid lesions and histology) serve to classify cases as LE or non-LE. (2) The 11 preliminary, dermatologic second step criteria classify LE cases as cutaneous LE or systemic LE. Interestingly, 5 of 11 of these second step criteria differ from the 11 ARA criteria for systemic LE. These second step criteria afford a useful means of distinguishing between subacute cutaneous LE cases with or without significant systemic involvement. CONCLUSIONS: The study factors included in both the first and the second step criteria fall into three groups, notably clinical criteria, laboratory criteria, and "added study factors." The latter factors distinguish between the groups compared (LE vs. non-LE and cutaneous vs. systemic LE) but not as well as the study factors included as "criteria." PMID- 8407092 TI - Prevalence of Raynaud's phenomenon and specific clinical signs related to progressive systemic sclerosis in the general population of Japan. AB - BACKGROUND: In order to evaluate the prevalence rates of Raynaud's phenomenon (RP) and specific clinical signs related to progressive systemic sclerosis (PSS) in the general population of Japan, inquiries were made concerning RP in the hands and dermatologic examinations were also conducted. METHODS: One thousand and sixty-three subjects (332 men and 731 women) over 30 years of age who underwent inhabitants' health examinations in 1990 were considered for this study. RESULTS: The prevalence of RP was 3.0% in men and 3.4% in women. In 8 men and 17 women with RP who received the blood tests, the positive rates of antinuclear antibody (ANA) were 12.5% and 35.3% in men and women, respectively. The prevalence rates of all five specific clinical signs related to PSS, sclerodactyly, pitting scars of the fingers, brown pigmentation of the body, shortened frenulum of the tongue, and flexion contracture of fingers, were under 2% in men and 3% in women. In women with RP the prevalence rates of sclerodactyly, pitting scars of the fingertips, brown pigmentation of the body, and shortened frenulum of the tongue were 16.0, 4.0, 4.0, and 16.0%, respectively. These values were significantly higher than those of persons without RP. CONCLUSIONS: Because some persons with primary RP may become typical cases of PSS within several years, a followup study, particularly for women who have positive titers of ANA with RP, should be carried out to find out whether the persons suffer from PSS or not. PMID- 8407093 TI - Role of emotional factors in adults with atopic dermatitis. AB - BACKGROUND: The role of anger in the onset or perpetuation of episodes of atopic dermatitis in adults has long been considered an important factor. The objective was to investigate whether atopic patients feel ineffective in dealing with anger and assertiveness when compared with psoriasis patients and control patients. METHODS: Thirty-four adult patients with atopic dermatitis were compared to 28 patients with psoriasis and 32 controls, dental patients without major skin disease. Standard measures of anxiety, anger, assertion, depression, and locus of control as well as a measure of anger effectiveness, designed for this study, were used. RESULTS: There were significant differences between atopic dermatitis patients and controls in that atopics felt angry more readily but were less likely to express it, were more anxious and less assertive, and felt less effective in expressing anger. The only difference between psoriasis patients and controls was less ability to express anger. Atopic patients were more chronically anxious than those with psoriasis. CONCLUSIONS: Adult atopic dermatitis patients are often chronically anxious and feel ineffective in handling anger which suggests that psychological interventions may prove helpful. PMID- 8407094 TI - Study of the partners of women with human papillomavirus infection. AB - BACKGROUND: Genital human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases, and it has been identified as a significant risk factor for the development of dysplasia and cancer of the uterine cervix. The possible influence of male HPV lesions on female cervix oncogenesis has not been elucidated so far. In the present study we evaluate the male partners of women with clinical or subclinical HPV infection with particular interest in the clinical features of this infection in both partners. METHODS: We examined 81 male partners of women affected with human papillomavirus infections. Condylomata acuminata were searched for by visual inspection. Subclinical lesions were searched by 5 power optical magnification lens after application of 5% acetic acid. RESULTS: In men we observed the following percentage of infection: 67% of the partners of women affected with condylomata acuminata, 46% of the partners of women affected with subclinical lesions (acetic acid positive), and 40% of the partners of women with association of HPV and cervical intraepithelial neoplasia. CONCLUSIONS: Our data stress that very often the partners of women with HPV subclinical infection, especially when associated with CIN, do not present lesions, and consequently primary prevention may be very difficult. PMID- 8407095 TI - Concurrent HPV-16 infection of the nipple and perianal area in an HIV-1+ patient. Military Medical Consortium for the Advancement of Retroviral Research (MMCARR) AB - BACKGROUND: In patients with HIV-1 disease there has been an increasing association with human papilloma virus (HPV) infections in multiple locations as well as an increase in associated tumors. In addition, there has been increased recovery of HPV in individuals with decreasing T4 cell counts. CASE REPORT: Recently we have seen an HIV-1+ patient with a cutaneous lesion on the nipple, as well as multiple perianal lesions in which HPV-16 was demonstrated by in-situ hybridization. Although these lesions contained the same subtype of HPV virus, they had very different clinical and histopathologic morphologies, and this represents the first reported association of HPV-16 in a nipple lesion. DISCUSSION: Our patient illustrates that in HIV-1 disease, HPV infections may present in more diffuse and atypical locations. In addition, the diffuse staining with the in-situ probe for HIV-16 within the lesions, tends to support the findings of others, that viral recovery increases with the immune suppression induced by HIV-1. PMID- 8407096 TI - Overheating in bed as an important factor in many common dermatoses. AB - BACKGROUND: Extensive questioning of patients with a wide variety of skin disorders led to the impression that nocturnal overheating was probably an important factor in the initiation and the perpetuation of many skin disorders. METHODS: In order to test the hypothesis, 12 "clean-skinned" subjects (6M/6F) aged 18 to 45 years were monitored electronically every 30 seconds during an 8 hour sleep period (2300 to 0700 hours), sleeping under a standard 10 tog duvet. RESULTS: All the subjects were too hot by 3 to 4 degrees C. All showed changes in their EEG patterns with reduced REM sleep, increased awakenings, and all showed changes in their sleep stage patterns. In addition, they all showed evidence of increased sweating in the "heat-sink" area. CONCLUSIONS: The mechanisms where by such changes could be implicated in the precipitation and perpetuation of skin disease are discussed. "Lifestyle" modification as a very effective, noninvasive, therapeutic regime is recommended. Further research along these lines would probably be very valuable and instructive. PMID- 8407097 TI - Janeway lesions: differential diagnosis with Osler's nodes. PMID- 8407098 TI - Pemphigus erythematosus induced by ceftazidime. PMID- 8407099 TI - Cutaneous squamous cell carcinoma in black patients with chronic discoid lupus erythematosus. PMID- 8407100 TI - Verrucous lesions of herpes simplex in HIV-1+ patients. Military Medical Consortium for the Advancement of Retroviral Research. PMID- 8407101 TI - Kaposi's sarcoma in an 18-year-old man. PMID- 8407102 TI - Treatment of alopecia areata with khellin and UVA. PMID- 8407103 TI - Podophyllin and pregnancy. PMID- 8407104 TI - Seasonality of presentation of keratoacanthoma. PMID- 8407105 TI - New method for the synthesis of N-methyl amino acids containing peptides by reductive methylation of amino groups on the solid phase. AB - Primary amino groups on the model peptide Xaa-Ala-Pro-Lys(ClZ)-Tyr(2BrZ), synthesized on a p-methylbenzhydryl amine resin with conventional Boc/benzyl protective group strategy, were reacted with 4,4'-dimethoxydityl chloride in dichloromethane, resulting in the introduction of the dimethoxydityl group, which is an acid-labile N-alkyl type of protective group. The secondary amino groups thereby formed can be methylated by treating the peptide-resin with formaldehyde and sodium cyanoborohydride in N,N-dimethylformamide. After the removal of the dimethoxydityl group with trifluoroacetic acid, the resulting N-methylated amino acid residues with a free secondary amino groups are accessible for acylation with the next activated Boc amino acid. With this method majority of the 20 common amino acids can be monomethylated directly on the resin and, in most cases, with very low levels of the side reactions. In the cases where the complete methylation is difficult to achieve, the remaining primary amino groups can be selectively acylated in the presence of secondary amino groups with trimethylacetic acid 1-hydroxybenzotriazole ester. The method provides a convenient general route to synthesize N-methylated derivatives of most of the occurring and synthetic amino acids. PMID- 8407106 TI - Vibrational spectroscopic studies of L,D-alternating phenylalanine peptides. AB - Infrared and Raman spectra of Boc-(D-Phe-L-Phe)n-OMe, with n = 4 and 5, and Boc-L Phe-(D-Phe-L-Phe)7-OMe, have been obtained in crystalline and solution states. Based on the earlier spectroscopic studies on Boc-(L-Val-D-Val)n-OMe, with n = 4, 6 and 8, and the predictions from the normal-mode calculations of double-stranded beta-helix conformations, structures have been assigned for the three oligopeptides. In the crystalline state, all three oligopeptides have a double stranded increases decreases beta 5.6 structure. Spectra from chloroform solution indicate that the solid-state conformation is retained in this solvent for all three oligopeptides. PMID- 8407107 TI - Studies on the carboxyl terminal peptides of human seminal plasma inhibin (HSPI). Chemical synthesis and in vivo biological activity of the disulfide loop peptide 67-94 of HSPI. AB - Observation of contradictory results with the in vitro assays for inhibin-like activity of the carboxyl terminal 28 amino acid peptide 67-94 with a disulfide loop, of human seminal plasma inhibin (HSPI), prompted us to synthesize both the linear and the cyclic peptides and test their ability to suppress the circulating levels of follicle stimulating hormone (FSH) in vivo in adult male rats. The linear peptide [Cys(Acm)73,87] 67-94 of HSPI was synthesized by solid-phase peptide synthesis using fluorenylmethyloxycarbonyl (Fmoc) chemistry and a continuous-flow technology. The peptide was cyclized by direct iodine oxidation of the S-diacetamidomethyl peptide in dilute solution. In the in vivo assay the linear peptide did not affect the levels of FSH, whereas the cyclic peptide suppressed the levels of FSH significantly. Thus, the carboxyl terminal region of HSPI does have inhibin-like activity and perhaps has the active core of the protein. PMID- 8407108 TI - Solid-phase synthesis of protected peptides using new cobalt(III) ammine linkers. AB - Cobalt(III) ammine complexes of the type cis-[CoL4(4-AMB)O-AA-Boc](CF3SO3)2, where L4 = bisethylenediamine (en)2 or tetraammine (NH3)4, and 4-AMB = 4 (aminomethyl)benzoic acid, have been synthesized and used as linkers to polystyrene resins for solid-phase synthesis of protected peptides. Boc/t-Bu protected [Leu5]enkephalin was assembled on the two different Co(III) resins, and then cleaved from the resins by reduction of the Co(III) center in 93-96% yield. HPLC-purified protected [Leu5]enkephalin was obtained in 67-69% overall yield and characterized by amino acid analysis and 1H NMR. Stepwise synthesis on the Co(en)2-resin was also used in the assembly of Boc-Asp(OcHex)-Arg(Mts)-Gly Asp(OcHex)-Ala-Pro-Lys(2Cl-Z)-Gl y-OH, a sequence from collagen alpha 1 Type 1. The protected peptide was cleaved from the Co(III) resin in 74% yield, and the HPLC-purified nonapeptide was characterized by amino acid analysis, 1H NMR and liquid secondary-ion mass spectrometry (LSIMS). New routes are described for the synthesis of isomerically pure Co(III) anchor complexes. The Co(III) resins were found to be compatible with both the tert-butyloxycarbonyl (Boc) and the 9 fluorenylmethoxycarbonyl (Fmoc) N alpha-protecting group strategies used in solid phase peptide synthesis. PMID- 8407109 TI - Synthesis and stability of 3-nitro-2-pyridinesulfenyl chloride (NpysCl). AB - 3-Nitro-2-pyridinesulfenyl chloride (NpysCl) is the starting material for the synthesis of N-, O- and S-Npys-protected amino acids. Two efficient, novel synthetic routes to NpysCl are described. The stability of NpysCl was determined in a variety of solvents, with and without base, to determine the most suitable solvent and base for the synthesis of N-Npys amino acids. The syntheses of Npys Ala and Boc-Lys(Npys) tert-butylammonium salt are also described. PMID- 8407110 TI - Solid-phase synthesis of O-mannosylated peptides: two strategies compared. AB - A comparison of the O-glycosylation of resin-bound assembled peptides with the incorporation of glycosylated amino acids using established chemistry is presented. Fmoc/tert-butyl-based protecting groups were used for the peptidic moieties in conjunction with acetyl sugar protection. Koenigs-Knorr glycosylations were carried out using protected bromomannose derivatives, the acceptor being threonine or serine, either in solution or within a resin-bound peptide. The characterisation of microgram quantities of glycopeptides by the use of glycosidases in combination with mass spectrometry is also described. PMID- 8407111 TI - Conformation-function relationships in LHRH analogs. I. Conformations of LHRH peptide backbone. AB - A systematic conformational build-up procedure was performed for the LHRH molecule, pGlu1-His2-Trp3-Ser4-Tyr5-Gly6-Leu7-Arg8-Pro9-Gly10-NH 2. The results showed a very high flexibility of the LHRH backbone, with 300 conformers being regarded as having low energy. At the same time, the conformational flexibility of LHRH differs among the fragments of the molecule. The most rigid fragments of LHRH are the Ser4-Tyr5-Gly6-Leu7 and Tyr5-Gly6-Leu7-Arg8 central tetrapeptides, the latter possessing only eight different types of low-energy backbone conformers. These eight conformer types belong to different kinds of chain reversals which are stabilized by different systems of intramolecular hydrogen bonds. Some of them resemble the beta-II' turn, which was derived as the LHRH structure from energy calculations by others. The results obtained are in good agreement with the experimental data on LHRH flexibility in solution. PMID- 8407112 TI - Conformation-function relationships in LHRH analogs. II. Conformations of LHRH peptide agonists and antagonists. AB - Systematic energy calculations were performed for a series of LHRH analogs including five agonists with substitutions of D- or N-Me-amino acid residues in positions 4, 6 and 7, and five antagonists with substitutions of D-, N-Me- or alpha-Me-amino acid residues in positions 1, 2, 3, 6, 7 and 10, as well as a bicyclic LHRH antagonist. The geometrical shapes of the calculated low-energy backbone structures for each compound were compared to those of LHRH itself. It appeared that the beta-II' turn at the Tyr5-Gly6-Leu7-Arg8 central tetrapeptide is the common structure for all LHRH agonists considered. LHRH antagonists also possess a common chain reversal in the central tetrapeptide, but it is different from that for LHRH agonists. The LHRH agonists share a similar low-energy conformer at the level of the entire peptide backbone. A characteristic feature of this conformer is a 'surface' formed by a 'polygon' with hydrophobic moieties of pGlu1, Trp3, Tyr5, Leu7 and Pro9 in the corners and with the side chain of the His2 residue in the middle, the latter being crucial for a manifestation of LHRH agonistic activity. Since the N-terminal tripeptide of LHRH presumably participates in a direct interaction with specific receptors, it is legitimate to suggest that the beta-II' turn in the central tetrapeptide maintains the proper spatial arrangement of the N-terminal tripeptide. On the other hand, LHRH antagonists considered in this study were shown to possess low-energy structures, with the spatial arrangement of the residues in the N-terminal tripeptide similar to that of agonists. This would suggest a new approach to the design of LHRH antagonists, namely by stabilizing this specific arrangement, rather than the beta-II' turn in the central tetrapeptide. PMID- 8407113 TI - Synthesis and solution conformation of c(D-Trp-D-Cys(SO3-Na+)-Pro-D-Val-Leu), a potent endothelin-A receptor antagonist. AB - The endothelin family of polypeptides are known to exert potent physiological effects which include cardiovascular regulation. The solution conformation and dynamics of c(D-Trp-D-Cys(SO3-Na+)-Pro-D-Val-Leu), a potent endothelin-A receptor selective antagonist, were characterized in aqueous solution by NMR spectroscopy and molecular modeling. NMR-derived conformational constraints were combined with computer-assisted molecular modeling using distance geometry calculations and energy minimization. The pentapeptide backbone is shown to adopt a single conformation in solution comprising a type II beta-turn and an inverse gamma turn, with each residue in the trans conformation. Molecular dynamics were explored using relaxation measurements and low-temperature studies, and indicate that the peptide backbone is highly constrained with little conformational mobility present. PMID- 8407114 TI - Multistep deprotection for peptide chemistry. AB - This review article focuses on some concepts which make use of special classes of protecting groups (protected protecting groups) in peptide synthesis. Terms such as gradative deprotection, dual deprotection, two-step deprotection, safety-catch protection and multidetachable protecting systems are discussed, and the general mechanistic considerations for the deprotection process are described. PMID- 8407115 TI - Detection of HIV in human vitreous. AB - Assay of human vitreous specimens obtained postmortem for HIV antibodies, or HIV p24 antigen, is reported to be a reliable technique to demonstrate HIV infection in possible cornea donors from whom serum could not be obtained. We tested three vitreous samples obtained during vitrectomy from two HIV-positive patients. One patient exhibited the clinical AIDS syndrome. HIV antigen and antibody tests were negative in all specimens. HIV proviral DNA was detected by PCR only in the vitreous of the patient with AIDS. Therefore, testing only vitreous samples is insufficient to exclude HIV infection in potential cornea donors. PMID- 8407116 TI - Surgical management of retinal detachment associated with optic nerve pit. AB - A patient with serous macular detachment associated with congenital optic nerve pit was successfully treated with pars plana vitrectomy, air-fluid exchange with internal submacular fluid drainage, endolaser, and sulfur hexafluoride (SF6) gas injection. PMID- 8407117 TI - Topical timolol maleate might adversely affect serum lipoproteins. AB - Conflicting observations have been reported about the effects of topically administered timolol maleate on serum lipoproteins. We therefore considered this issue in a series of eight glaucoma patients receiving timolol maleate. Cholesterol and triglycerides were measured in plasma and in low-density lipoproteins (LDL), and high-density lipoproteins (HDL), both before and following three months of treatment. Following the treatment, the mean atherogenic index was increased from 2.72 to 3.38 (p = 0.012). This suggests that the atherogenic index should be determined before and during timolol maleate treatment in high-risk cardiovascular patients. PMID- 8407118 TI - A combined anterior and posterior approach to cataract surgery in patients with chronic uveitis. AB - Although several recent papers have focused on the results of cataract surgery in patients with uveitis, little has been published on specific surgical techniques that are most appropriate to such cases. We have found that a combined anterior and posterior approach using extracapsular techniques (usually phacoemulsification) and pars plana vitrectomy, with or without intraocular lens placement, is best suited for selected uveitis patients who have cataract, vitreous opacities, and cystoid macula edema. This paper discusses the surgical techniques that we have found to be best for the management of these cases. PMID- 8407119 TI - Classification of 750 consecutive uveitis patients in the Rotterdam Eye Hospital. AB - The data of 750 consecutive uveitis patients who visited the uveitis department of the Rotterdam Eye Hospital were analysed by computer. Anterior uveitis was diagnosed most frequently (52%), mostly the acute form. We found a high percentage of Fuchs' heterochromic iridocyclitis and HLA-B27+ iridocyclitis. In the posterior uveitis group appeared the more recent clinical entities such as acute retinal necrosis, Birdshot retinochoroidopathy, CMV-retinitis, AMPPE and ocular candidiasis. Ocular Toxoplasmosis was still the leading cause in this group. These data will be compared with other studies of similar populations. Factors influencing frequency of subtypes of uveitis and bias of this study will be discussed. PMID- 8407120 TI - Use of vancomycin in vitrectomy infusion solution and evaluation of retinal toxicity. AB - The retinal toxicity of vancomycin in infusion solution used in vitrectomy and lensectomy was investigated in rabbit eyes by means of electroretinography and histologic study (light microscopy). Concentrations of 8 micrograms/ml, 16 micrograms/ml, and 32 micrograms/ml of vancomycin in balanced salt solution caused no abnormal ERG or histologic changes. However, ERG amplitude depression and abnormal histologic changes occurred when the concentration of 100 micrograms/ml of vancomycin was used. PMID- 8407121 TI - Effect of contact diode laser on the cornea with and without absorbing dye. AB - The semiconductor diode laser is a near-infrared laser; its 810-nm wavelength is maximally absorbed by melanin and has substantial transmissibility through cornea and sclera. Indocyanine green is the best photosensitive dye for the diode laser. The level of corneal damage produced by 810-nm diode laser, with and without absorbing dye (indocyanine green), and photoablative capabilities of this wavelength were studied using albino rabbits. We concluded that the contact application of this wavelength to the cornea in the presence of energy-absorbing dye causes both stromal and endothelial thermal damage. Therefore, 810-nm near infrared semiconductor diode lasers are not suitable for photorefractive keratectomy or photoablative reprofiling. PMID- 8407123 TI - Convergences and divergences in contemporary psychoanalytic technique. AB - A broad survey of the psychoanalytic field reveals both convergences and divergences in technique. The major convergences include earlier interpretation of the transference, increased focus on transference analysis, as well as growing attention to countertransference analysis and increasing concern with the risks of 'indoctrinating' patients. Greater emphasis is found on character defences and the unconscious meanings of the 'here-and-now'. Also noted are trends toward translating unconscious conflicts into object-relations terminology, as well as toward considering a multiplicity of royal roads to the unconscious. Regarding divergences, significant controversies continue about the importance of the 'real' relationship, and the therapeutic versus the resistance aspect of regression. Divergences also continue regarding reconstruction and recovery of preverbal experience, drawing the lines between psychoanalytic psychotherapy and psychoanalysis, the role of empathy, and the relation of historical to narrative truth. PMID- 8407122 TI - The use of tissue plasminogen activator in postvitrectomy cases. AB - The study concerns 17 eyes which, following vitrectomy, were given an injection of 25 micrograms of tissue plasminogen activator (tPA). Of these 17 cases, ten showed a severe fibrin formation in the anterior chamber, 3 cases showed vitreous hemorrhage (one of them with hyphema), 2 had fibrin formation and cellular proliferation, while in one case tPA was injected at the end of the vitrectomy because of perisilicone proliferation and in one case because of fibrin depositions on the intraocular lens. The tPA was injected into the anterior chamber (10 eyes) or into the vitreous cavity (7 eyes). The follow-up period ranged from 4 to 15 months (mean period 9 1/2 months). Fibrinolysis was noted in the 10 cases with fibrin formation in the anterior chamber. Fibrin dissolution was achieved within 3-4 hours. None of these cases presented a recurrence throughout the follow-up period. Positive results were observed also in the case with perisilicone proliferation. On the contrary in 3 cases with postvitrectomy hemorrhage the hemorrhage persisted unchanged. Also in 2 cases with fibrin formation and cellular proliferation on the anterior and posterior surface of the iris the tPA injection proved ineffective. Both cases developed traction retinal detachment (TRD) due to anterior proliferative vitreoretinopathy (PVR). In the case with fibrin depositions on the intraocular lens the situation remained unchanged. Any complications observed in our case proved to be mild and transitory. PMID- 8407124 TI - Disruptions of the therapeutic relationship in psychoanalysis: a view from self psychology. AB - The progression of the therapeutic process in clinical psychoanalysis is, characteristically, punctuated by a series of breaches in the otherwise collaborative analytic relationship of analyst and analysand. In this paper, these disruptions are conceptualised from the point of view of self psychology. A focus on the subjective aspects of the analytic relationship, i.e. the selfobject experiences in the transference, illuminates the vicissitudes of the emergence, maintenance and transformation of the self. The analytic process proceeds against an unconscious conflict between the fears of repeated injury and the longing for needed selfobject experiences. Disruptions occur when analysands perceive the analyst to be less attuned to their issues and more involved in the analyst's own agenda. Acknowledgement of the analysand's subjective view as a valid experience must precede the interpretation of the relational dynamics before it can effectively lead to a restoration of the process. PMID- 8407125 TI - Deficiency and envy: some factors impacting the analytic mind from listening to interpretation. AB - This paper examines some aspects of an unconscious process developing within the mind of the analyst; a process affecting both the way in which the analyst listens to the patient's material and the interpretations subsequently formulated. The paper highlights the problem of the analyst's 'mis-use' of theory as a protection against unbearable feelings of psychic pain, evoked by what is perhaps the most essential tool of psychoanalysis: the analyst's emotional contact with the patient's early experiences in infancy. Both the 'deficiency theory' and the concept of 'primary envy' are called into question during this inquiry. Some of the most recent work of the 'post-Kleinians (from Bion through to the present-day workers in England as well as in the Americas) is reviewed. Clinical examples, both from the author's own experience and from previously published material, are considered as illustrative of some factors which may impact upon the mind of the psychoanalyst from listening and interpretation. PMID- 8407126 TI - The cracked container, the containing crack: chronic illness--its effect on the therapist and the therapeutic process. AB - In this paper, we have tried to examine the complex dynamics of the therapist's physical chronic illness. Little attention has been paid to physical non functioning, especially during chronic illness, and its psychotherapeutic corollaries. A crucial theme is the extent of damage to the capacity to contain. It is our claim that, when sufficiently worked through, the 'cracked therapeutic container' may serve as a facilitator of better understanding and enhance empathy. The impact of chronic illness on the therapeutic setting, contract, process and language is discussed as well as some salient features of chronicity. PMID- 8407127 TI - Psychoanalysis and maternal work--some parallels. AB - This paper makes a comparison between a certain type of dilemma facing the analyst in his daily decisions about how to intervene and what to interpret, and the dilemmas faced by parents concerning what would be most useful, at a particular moment, in facilitating the growth of their children. Both analytic and parental strategies move back and forth between emphatic closeness and support, on the one hand, and more objective understanding--involving a 'vision of the future'--and facilitating individuation, on the other. The manner in which these strategies are used is inherently dialectic, and, one hopes, suited to the child's or patient's needs, but also depends on the style of the analyst. This similarity between the work of analysts and that of parents (assuming, of course, the basic differences between these two vocations!) is congruent with the growing prevalence, in almost all contemporary psychoanalytic thought, of object relations theory. PMID- 8407128 TI - Beneath the analytic surface: the analysand's theory of mind. AB - A growing body of research on the way children change their forms of thinking and views of the world from around the ages of 2 to 5 strongly suggests that something called a 'theory of mind' is acquired during this time which significantly affects the course of language acquisition, moral development, interactive problem-solving and other cognitive attributes. As we learn more about the adult varieties of this theory of mind we begin to realise their implications for psychoanalysis. This paper considers such issues as: what kind of theory of mind is assumed by our reliance on the Basic Rule? What kind of theory is required to understand the usual range of standard interventions? How can deficiencies in the analysand's theory of mind interfere with standard interpretations? As research on theory of mind makes us increasingly sensitive to the importance of different forms of thinking about thinking, we can apply many of these insights to common problems of psychoanalytic technique. PMID- 8407129 TI - An interpretation of transference. AB - This paper offers an interpretation of the concept of transference. It begins with a re-examination of Socrates's therapeutic intent and Plato's revisions of psychological theory in the light of Socrates's failure. In particular, Plato devised an account of a structured psyche, a theory of fantasy, and a complex 'object-relations' theory, to explain dynamic interactions between psyche and social world. It is argued that these revisions embody a recognition of the phenomenon of transference, i.e. the psyche's characteristic activity of creating a meaningful world in which to live. Freud's early conception of transference--as a transfer of psychological content across space and time--is criticised on the ground that it assumes that the world is already given, independent of any psychic activity. Freud's later conception of transference as a repetition is then explored in the light of the psyche's ability to create artefacts. It is argued that neurotic transference is the unconscious attempt to create an idosyncratic polis, an idiopolis, in which to live. The resolution of the transference occurs because the analyst is made a citizen of the idiopolis, has learned the idiolect, and can speak in that idiolect of the fundamental conflicts within the idiopolis and their dynamic basis. A neurotic world cannot survive this internal recognition. PMID- 8407130 TI - Transference, countertransference and acting out in psychoanalysis. AB - The development of the transference concept is described and studied from structural, genetic, dynamic and adaptive points of view. A sketch is given of the development of a transference neurosis during treatment, and of the difference between transference phenomena and transference neurosis. A number of examples from practice are given, also in connection with the occurring repetition compulsion. In the discussion of the countertransference concept a distinction is made between the so-called classic approach and the totalistic approach to this concept. The author studies the possible interference of the countertransference during treatment. If the analyst can withdraw from his trial identification with his patient no interference will occur. Sudden countertransference phenomena may be eliminated by self-analysis. Chronic countertransference phenomena can only be solved by re-analysis. Examples from practice are given and the fact is demonstrated that analyst and patient are not only transference-countertransference objects for each other, but also real objects. Attention is paid to the old and the new concept of acting out. Examples from practice are given of acting out during and outside the analytic sessions. PMID- 8407131 TI - 'Through privation to knowledge': unknown documents from Freud's university years. AB - The paper is centred on three hitherto unknown grants, two of them donated by different Jewish Foundations to the medical student Freud shortly before the end of his studies. It furthermore turned out that Freud's salary as a demonstrator in Brucke's Laboratory had been a grant too, donated by the University of Vienna. Aspects of the contemporary background by means of the private foundations and their donors as well as of the aspiring tendencies at the University of Vienna are described. Documentary evidence demonstrates that Brucke promoted Sigmund Freud continuously, more than hitherto known in the biography. The significance of Brucke and Charcot as identification-figures, on the one hand, and for Freud's development as a scientist, on the other, is elaborated. Contemporary quotations shed light on Freud's poverty in his youth as well as the arising anti-Semitism at the University of Vienna. PMID- 8407132 TI - Splitting of the ego, as the central phenomenon in neurosis. Ritske le Coultre. 1966. AB - The phenomenon of a split in the ego is not confined to fetishism, but is central to all neuroses. The author demonstrates this with regard to several specific neurotic types. He then goes on to show that the attempt to establish an absolute distinction between ego and id postulates an artificial split between--in fact- mutually interdependent parts. Traditional distinctions between primary and secondary process are critically examined. The id is an abstraction, while the essence and strength of the ego reside not in its defensive functions, but in the extent to which it accepts the drives; whereas the repressing ego weakens or even eliminates itself. The child who cannot resolve psychosexual conflict uses a makeshift solution, whereby that portion of the ego engaged in the conflict is split off and ceases to participate in further development. This part, emotionally rich, may again blossom and mature in the transference. PMID- 8407133 TI - Curative factors in the psychoanalyses of Holocaust survivors' offspring before and during the Gulf War. AB - In this paper, I have attempted to explore the curative effect of insight and relational factors in the analyses of Holocaust survivors' offspring before and during the Gulf War. A particular characteristic of children of survivors is their tendency to recreate their parents' experiences in their own life through concretization. An important analytic goal is to help these patients become aware of the unconscious meaning embedded in their acting out through increased insight, so they will be able to extricate themselves from the need to concretize and verbalize instead. The impact of the Gulf War on the children of Holocaust survivors was particularly strong. These patients reacted to the existential threat with feelings of impotence and terror, perceiving it as a repetition of the past. Thus, strengthening the ego forces became the focus of treatment during the war period, and this was facilitated by relational factors. Only near the end of the war was it possible to begin working through the regressive transferences evoked by the traumatic situation through increased insight, or to attempt to disentangle the present from the past through interpretation. PMID- 8407134 TI - Some dream mechanisms in Finnegans Wake. AB - This paper does not purport to offer a psychoanalytic reading of Finnegans Wake, but rather to demonstrate how, in recreating the mechanisms of the dream, Joyce's masterpiece offers to psychoanalysis a uniquely rich opportunity to explore the shadowy play of the dream in the permanence of a work of art. In particular, the manner in which Finnegans Wake tells its story through the distortions of dream narrative while bodying forth its protagonists in the substitutions of dream identity are explored, with the help of certain Freudian and Lacanian concepts of unconscious structure. PMID- 8407135 TI - John Bowlby (1907-1990). PMID- 8407136 TI - Angel Garma (1904-1993). PMID- 8407137 TI - Freud's oratorical marathon: the shrinking of another myth. PMID- 8407138 TI - Psychotic behavior associated with cerebellar pathology. AB - This report concerns a 27-year-old man in whom psychotic behavior emerged in association with a cerebellar tumor. This presentation supports previous clinical observations which postulate an intimate relationship between cerebellar pathology and the development of schizophrenia. PMID- 8407139 TI - The cognitive-mnestic performance profile of a patient with bilateral asymmetrical thalamic infarction. AB - The case of a 46-year old patient with a bilateral medial thalamic infarct, affecting mainly the region of the internal medullary lamina, but not of the mammillothalamic tract, is documented neuropsychologically and neuroradiologically. Testing was done after acute remission. The patient was of average intelligence but manifested a number of verbal memory problems. These are most likely attributable to the selective diencephalic damage and are interpreted as a disconnection syndrome. PMID- 8407140 TI - Amphetamine-induced rotational behavior in rats: relationship to hypothalamic and striatal degeneration. AB - When lesions are placed unilaterally in the nigrostriatal system of experimental animals, rotational behavior occurs in response to peripheral administration of dopamine (DA) agonists. In spite of considerable evidence to the contrary, it is assumed that in order for this rotation to occur, an almost complete depletion of striatal DA must be achieved. To test this hypothesis further, 20 male Sprague Dawley rats were injected unilaterally with 2 microL of 8 micrograms/microL of 6 hydroxydopamine (6-OHDA) via acute injection needles or chronically indwelling cannulae. Acute injection of 6-OHDA resulted in a rotation rate of 7.2 to 18.9 revolutions per minute in response to peripheral amphetamine injection (5 mg/Kg) while injection of 6-OHDA through chronically indwelling cannulae produced rotation ranging from 1.4 to 9.9 rotations per minute. Under the conditions of either method of injection, the animals displaying the most severe rotation still showed partial denervation of striatal DA as revealed by catecholamine fluorescence histochemistry. Conversely, numerous animals demonstrating very low rates of amphetamine-induced rotation often displayed a complete loss of striatal, accumbens, and olfactory tubercle catecholamine fluorescence. Moreover, large quantities of lateral hypothalamic amine accumulation were observed in rotating rats indicating that this neurochemical change may be of functional significance for rotational responses. The present results, when taken into consideration with previous work, indicate that the routine selection of rotating animals for pharmacological testing for potential antiParkinsonian medication or intracerebral grafting purely on the basis of their rotational behavior does not necessarily imply that complete striatal denervation has occurred. Moreover, these findings demonstrate that amine accumulation in the lateral hypothalamus of rotating animals with DA depleting lesions is an important phenomenon implicated in the expression of rotational behavior in animals and possibly in the pathophysiology of Parkinson's disease. PMID- 8407141 TI - Multiple sclerosis: relationship between seasonal variations of relapse and age of onset. AB - The incidence of multiple sclerosis (MS) is age-dependent being rare prior to age 10, unusual prior to age 15, with a peak in the mid 20s. The manifestation of MS, therefore, appears to be dependent upon having passed through the pubertal period suggesting an endocrine influence on the timing of onset of the disease. Since pineal melatonin secretion progressively declines from childhood to puberty and as melatonin exerts an immunomodulating influence, we have proposed that the dramatic decline in melatonin secretion just prior to the onset of the clinical manifestations of puberty may lead to disruption of immune responses resulting in either reactivation of the infective agent or in an increased susceptibility to pubertal or post-pubertal infection. Melatonin secretion undergoes annual rhythms with a zenith in winter and declines to a nadir in the spring. Thus, the fall in melatonin secretion in the spring may account for epidemiological findings revealing a high incidence of relapse of MS in the spring. If the manifestations of MS are related to the fall in melatonin secretion in the post-pubertal period, then one would expect patients with a pubertal onset of the disease to have a higher incidence of relapses in spring than in winter. To test this hypothesis, we investigated in 51 patients the relationship between the seasonal occurrence of the last MS relapse with the age of onset of first manifestation of MS. While 9 of 22 patients (40.9%) who relapsed in spring (March-May) had the onset of MS prior to age 18, only 2 of 29 patients (6.9%) who relapsed in winter (November February) experienced the onset of first symptoms prior to the age of 18 years (p < .005). These findings thus support the hypothesis implicating the pineal gland and melatonin secretion in the timing of onset of MS. Moreover, the findings may have clinical implications with respect to the prophylaxis of MS relapse in patients who experience seasonally-dependent exacerbation of symptoms. PMID- 8407142 TI - Electrophysiological comparison between catechol- and urea-induced myoclonus models in the rat. AB - Catechol- and urea-induced myoclonus models in the rat were electrophysiologically compared to clarify pathophysiological differences. Catechol-induced myoclonus had various similarities with cortical reflex myoclonus in that there were electroencephalogram (EEG) discharges prior to myoclonic discharges, a spread of myoclonic discharges from the rostral to the caudal site, and a high amplitude somatosensory evoked potential (SEP). In urea induced myoclonus, there were no EEG discharges related to myoclonic discharges and no enlarged SEP components as in reticular reflex myoclonus. Catechol-induced myoclonus had two evoked EMG responses of the biceps femoris at mean onsets of 8.0(C1) and 13.4 (C2) ms, and urea-induced myoclonus had a response (U1) at the mean onset of 10.2 ms. A study of the effects of various lesions in the central nervous system on these evoked EMG responses suggests that C1 is a monosynaptic spinal reflex. C2 which disappeared when the bilateral sensorimotor cortex for the hind limb had been resected and the lesion cooled is generated by the deep cerebral structures, such as the thalamus or basal ganglia, and U1 originates in the brain stem reticular formation. These results imply definitive differences of the pathophysiological mechanisms between catechol- and urea-induced myoclonus. PMID- 8407143 TI - Nocturnal melatonin secretion in suicidal patients with multiple sclerosis. AB - Multiple sclerosis (MS) is characterised by the occurrence of patchy CNS demyelinating lesions, leading to various degrees of motor, sensory, affective, and cognitive deficits. MS is associated also with an increased risk of suicide accounting for a substantial rate of death among these patients. Post-mortem studies in suicide victims with various psychiatric disorders demonstrate a decreased concentration of serotonin (5-HT) and its metabolites in the brain. Since 5-HT is a precursor in the synthesis of melatonin and as pineal melatonin content was found to be low in suicide victims, we predicted lower melatonin secretion in suicidal versus non-suicidal MS patients during an acute exacerbation of symptoms. To test this hypothesis, we investigated nocturnal plasma melatonin levels in a cohort of 28 relapsing patients who were admitted consecutively to an inpatient Neurology service, 6 of whom had a history of suicide attempts and were having suicidal ideation at the time of admission. While both cohorts of patients were not distinguishable on any of the demographic data including use of psychotrophic drugs on the day of admission to hospital, the mean melatonin level in the suicidal group was significantly lower than in the control group (19.0 pg/ml +/- 11.9 versus 45.5 pg/ml +/- 27.1; p < .05). These findings support the prediction of the study implicating the pineal gland in the pathogenesis of suicidality in MS and reinforce the concept that a biological rather than a reactive etiology underlies the development of psychiatric symptoms in MS. PMID- 8407144 TI - Postpolio syndrome--response to deprenyl (selegiline). AB - Two patients with postpolio syndrome are presented. The first case developed mild Parkinson's syndrome, for which she was treated with a levodopa/carbidopa combination followed by the institution of deprenyl. An unexpected improvement in the symptoms of postpolio syndrome was noted. The second patient who was unaffected by Parkinson's syndrome was started on deprenyl alone and reported a similar improvement in symptomatology. PMID- 8407145 TI - Inverse relationship between nonverbal intelligence and the parameters of pattern reversal visual evoked potentials in left-handed male subjects: importance of right brain and testosterone. AB - The relationships between latencies of visual evoked potentials (VEPs) and nonverbal intelligence test scores (IQs from Cattell's Culture Fair Intelligence Test) and correlations between serum testosterone level and VEP latencies were studied to examine the neural speed hypothesis of intelligence and its hormonal mechanisms in left-handed male subjects (Geschwind Scores). In accordance with the speed theory of intelligence, N1 and P1 latencies (and amplitudes) were found to be inversely related to IQ. However, this was true only for the right brain; the left brain did not contribute to this relationship. There was an inverse relationship between serum testosterone level and P1 latency; left minus right N1 latency, depending on N1 latency from right brain, linearly increased with testosterone. It was concluded that nonverbal intelligence largely depends on speed of information processing only by the right brain, not by the left brain in left-handed male subjects. This may have been created by testosterone in these subjects. Inconsistencies in the IQ literature concerning the speed hypothesis of intelligence may be explained by differences in cerebral lateralization. PMID- 8407146 TI - Information processing and interhemispheric transfer in left- and right-handed adults. AB - Two experiments are reported using simple reaction time (SRT) and visual evoked potentials (VEP) to measure information processing and interhemispheric transfer time (IHTT) in left- and right-handed subjects. Study 1 used an SRT procedure to estimate IHTT in 30 left-handed and 30 right-handed subjects. No differences in IHTT were indicated, but left- and right-handers were found to have opposite visual field advantages on the SRT task, indicating possible hemispheric differences in visual information processing. Study 2 was designed to test hypotheses generated in Study 1, using a VEP procedure to provide additional measures of visual information processing and IHTT. VEPs were collected during an SRT procedure in 20 left-handers and 20 right-handers, and SRT response hand and visual field advantages were replicated among right-handers only. Left-handers in Study 2 demonstrated no significant response hand or visual field effects on SRT. Analysis of the VEP N160 latency data indicated no significant laterality, hemisphere, or visual field effects, except for a robust hemisphere by visual field interaction, which reflects IHTT. The significant SRT left visual field advantage in right-handers was not reflected by hemispheric asymmetries in VEP N160 latency. It is concluded that left-handers and right-handers do not differ in IHTT in two functional regions of the corpus callosum, nor do they vary in simple visual receptive processing. Results are more consistent with variability in SRT resulting from differences in complex heteromodal processes beyond the striate and extrastriate visual cortex. PMID- 8407147 TI - Relationships among nonverbal intelligence, hand speed, and serum testosterone level in left-handed male subjects. AB - The relationships among nonverbal intelligence, hand speed, and serum testosterone level were studied in male left-handers ranging in age from 17 to 19 years. Hand speed was measured by a peg moving task. To assess the differences between nonverbal IQs. Cattell's Culture Fair Intelligence Test was used. There was a direct correlation between IQ and testosterone. IQ increased linearly with right-hand speed, which was directly related to testosterone. There was no significant correlation between IQ and left-hand speed, which was not significantly correlated with testosterone. IQ decreased with left- minus right hand speed, which also decreased with testosterone. It was suggested that nonverbal spatial reasoning ability may be directly associated with the efficiency of left brain, which is favored by testosterone in male left-handers. It was also concluded that the left to right asymmetry in hand speed may depend on efficiency of the right brain in left-handed males. PMID- 8407148 TI - Rhythmic variations of P100 component of pattern-reversal visual evoked potentials. AB - Pattern-reversal visual evoked potentials (EPs) were recorded in seven adult subjects to assess the existence of rhythmic variations in processing visual information at a primary stage. During a session of two hours, sixty EPs were recorded on the left and right cerebral hemispheres. The amplitude of N75-P100 component was measured. Remarkable variations were found with a periodicity from 15 to 60 min in six out of seven subjects. No hemispheric differences were found in the rhythmic variations. The results are in agreement with the behavioral data which show ultradian variations in visual performance depending on a general activation oscillator. PMID- 8407149 TI - Immunomodulation by conditioning: recent developments. PMID- 8407150 TI - A multiple modality approach combining the effect of conditioning with adoptive chemoimmunotherapy. AB - It has been demonstrated that significant protection against YC8 lymphoma can be induced in mice preimmunized with normal DBA/2 spleen cells. The DBA/2 spleen cells used as an alloantigen share minor histocompatibility determinants with the YC8 tumor. Our observations showed that once tumor was present in vivo, the use of a potent tumor specific vaccine that can confer 100% protection to preimmunized animals, can help in increasing survival but can no longer produce high incidence of regression and cure. We have used this model to show that adoptive chemoimmunotherapy (ACIT) can be used to regress tumors in mice with large body burden of tumor and that combination of conditioning with ACIT appears to enhance the effectiveness of treatment. The nature of the immunity conferred by conditioned resistance might be due to cytotoxic T-lymphocytes. PMID- 8407151 TI - Short and medium-term influence of physical activity on immune parameters. AB - Physical stress induces different changes in immune parameters depending on effort schedule and/or customary physical training. The mechanisms whereby they take place and the occurrence of possible tolerance after repeated effort have not been conclusively elucidated. We studied short and medium-term exercise induced changes in immune parameters after a standard physical effort (24' of cycle ergometer up to the 80% of maximal heart rate, daily for 5 days) in a group of healthy untrained controls. White Blood Cells, lymphocyte subsets, plasmatic catecholamine and cortisol levels, IgG and IL2receptor (IL2R) levels were determined. Most of the observed changes were strictly acute effort-related and disappeared within 3 hours (except for shifts in CD4+ CD45RA+ and CD4+ CD45RA- lymphocytes): they were concomitant to a transient sympathetic activation proved by heart rate (HR) and Norepinephrine (NE) increase. The medium-term effects of repeated daily effort included only a questionable rise in CD19+ and CD3+ CD4- CD8- cells. As far as possible tolerance mechanisms are concerned, we did not detect any change in either the direction or the entity of effort-induced changes in our controls after repeated effort. Study of strictly standardized exercise protocols is mandatory before clinical applications of physical activity in the approach to the treatment of disimmune diseases. PMID- 8407152 TI - The Lyapunov exponent of heart rate dynamics as a sensitive marker of central autonomic organization: an exemplary study of early multiple sclerosis. AB - In a study of central autonomic dynamics in early multiple sclerosis, we measured the temporal oscillations of the momentary heart rate (heart rate dynamics). 11 young patients suffering from relapsing remitting definite multiple sclerosis in relapse-free and early stages of illness and 11 healthy controls were examined under vagotonic and sympathicotonic conditions. The temporal structure of the heart rate dynamics was operationalized phase-space analytically through the estimation of the largest Lyapunov exponent. Positive Lyapunov exponents were found in all participants under all conditions indicating deterministically chaotic heart rate oscillations. The variance analysis of these exponents detected no significant effect of sympathetic or vagal activity (experimental condition) but a significant group difference (p < .02). The multiple sclerosis patients were characterized by significantly lower Lyapunov exponents than the healthy controls. This finding suggests a more stable and thus less adaptive central-autonomic organization in early multiple sclerosis. PMID- 8407153 TI - Lateralized lexical decisions and the effects of hemifield masks: a study of interhemispheric inhibition and release. AB - A lateralized tachistoscope experiment using 40 normal, right-handed men tested the predictions of three different theories of interhemispheric interactions. Lexical decision for function words was measured for each visual field under several conditions of simultaneous stimulation in the contralateral hemifield. When a nonsense word was presented simultaneously to the contralateral hemifield, performance decreased, especially for words in the weaker field, an effect which is inconsistent with the predictions of the direct access model. When the contralateral nonsense word was immediately masked, there was a significant improvement in performance over the nonmasked condition. Analysis of the masking effect showed that the data are inconsistent with a callosal relay model, but are consistent with a model of transcallosal inhibition. PMID- 8407154 TI - Dichotic listening and the cerebral organization of the phonetic and semantic components of language. AB - Two words were presented dichoticly and the subject was required to point to visually presented words that either rhymed with or were semantically related to those words. Task demands were varied in terms of the number and type of stimuli which the auditory and visual words actually matched. Performance was superior when either type of stimulus was presented to the Left Hemisphere via the Right Ear, resulting in the standard Right Ear Advantage (REA) for verbal material. The strength of the REA, however, depended both on the type of stimuli and the specific demands of the task. It was concluded that, since language has many components, dichotic listening can be used to investigate the relative contributions of the two cerebral hemispheres to those various components. PMID- 8407155 TI - Time-of-day and attentional-order influences on dichotic processing of digits in learning disabled and normal achieving children. AB - A heterogeneous group of 26 learning disabled (LD) and 30 normal achieving (NA) children, responded to a dichotic listening task using digits in morning and afternoon settings. Attentional order (i.e., right ear first versus left ear first) interacted with (1) Time-of-Day and (2) Group and Ear Attended. The first interaction revealed, as predicted, higher morning performance for subjects directed to attend right first. Subjects directed left first showed higher afternoon performance. These results are consistent with enhanced left hemisphere involvement after left hemisphere priming in the morning, and after right hemisphere priming in the afternoon. The second interaction indicated the LD had more difficulty than controls switching attention to the right ear when instructed to attend left first. The LD may activate the right hemisphere (via left hemispace attending) and have difficulty with subsequent right hemisphere inhibition, or left hemisphere activation, when shifting to right ear attending. Nonparametric tests revealed a greater incidence of lateralized responders in the morning for normal achievers attending left first. Findings are seen to augment previous research. PMID- 8407156 TI - Cerebral dominance, sleep and dream phenomena. AB - Evidence from human and animal studies is discussed in relation to the role played by the cerebral hemispheres in the genesis of sleep and particularly dream phenomena. The inconclusiveness of such research is commented upon and suggestions for further research are outlined. PMID- 8407157 TI - Modification of activation and evaluation properties of narratives by weak complex magnetic field patterns that simulate limbic burst firing. AB - In two separate experiments a total of 71 volunteers were asked to generate spontaneous narratives that were scored automatically by the Whissell Dictionary of Affect. During the narratives, weak (1 microT; 10 mG) magnetic fields were applied briefly through the temporal planes. In Experiment I, subjects who were exposed to simple sine wave or pulsed fields generated more scorable words that indicated lower activation and evaluation than sham-field conditions. In Experiment II subjects exposed to a computer-generated wave form, designed to simulate neuronal burst firing, generated narratives dominated by more pleasantness and less activation than a reference group. The possibility that this approach could be utilized to study the affective dimension of language selection was indicated. PMID- 8407158 TI - Choroid plexus calcification as a possible marker of hallucinations in schizophrenia. AB - Several studies suggest that disturbances of serotonin (5-HT) functions may be involved in the pathophysiology of hallucinations in schizophrenia. It is now well established that the choroid plexus (CP) is innervated by serotonin (5-HT) neurons, which may regulate its activity, and it is possible that decreased 5-HT functions may facilitate the process of its calcification. It is thus conceivable that calcification of the CP may be associated with hallucinations in schizophrenia. I studied in 18 chronic schizophrenic patients the association of CP calcification (CPC) size as ascertained from CT scan, to severity of hallucinations and, for comparison, to four other positive symptoms as well as global psychopathology score. Analysis of variance indicated that CPC size was specifically associated with hallucinations (p < .001) and none of the other psychopathology measures. These findings reveal a relationship between CPC and hallucinations in schizophrenia and suggest that the former may be a neuroradiological marker of hallucinations in the disease. PMID- 8407159 TI - Late-onset schizophrenia: relationship to awareness of abnormal involuntary movements and tobacco addiction. AB - Although schizophrenia usually emerges at mid to late-adolescence, it has been estimated that almost 20% of schizophrenic patients develop their first symptoms in mid to late life (late-onset schizophrenia). The biological characteristics that distinguish patients with early onset from those with late-onset schizophrenia have not been well delineated. A subgroup of neuroleptic-treated schizophrenic patients develops tardive dyskinesia (TD) and the majority of these patients are unaware of their movements. To investigate whether early and late onset schizophrenic patients with TD could be differentiated on the basis of awareness of involuntary movements, we compared the prevalence of awareness of these abnormal movements in patients with early (N = 40) and late-onset (N = 15) schizophrenia. We found a significantly higher prevalence of awareness of involuntary movements in patients with late-onset schizophrenia as compared to those with an earlier age of onset (86.6% vs. 25.0%, p < .0001). In a second study, we investigated whether early and late-onset schizophrenia could be differentiated on the basis of dopamine functions in the mesolimbic system. Since tobacco addiction is mediated via limbic dopaminergic functions, we investigated the prevalence of tobacco addiction in patients with early (N = 51) and late onset (N = 13) schizophrenia. We found a significantly lower prevalence of tobacco addiction in patients with late-onset schizophrenia as compared to those with an earlier age of onset (15.4% vs. 54.9%; X2 = 6.49; p < .01). Our findings support the notion that distinct pathophysiological mechanisms underlie the development of early and late-onset schizophrenia. PMID- 8407160 TI - Vitamin B12 and its relationship to age of onset of multiple sclerosis. AB - Attention has been focused recently on the association between vitamin B12 metabolism and the pathogenesis of multiple sclerosis (MS). Several recent reports have documented vitamin B12 deficiency in patients with MS. The etiology of this deficiency in MS is unknown. The majority of these patients do not have pernicious anemia and serum levels of the vitamin are unrelated to the course or chronicity of the disease. Moreover, vitamin B12 does not reverse the associated macrocytic anemia nor are the neurological deficits of MS improved following supplementation with vitamin B12. It has been suggested that vitamin B12 deficiency may render the patient more vulnerable to the putative viral and/or immunologic mechanisms widely suspected in MS. In the present communication, we report that serum vitamin B12 levels in MS patients are related to the age of onset of the disease. Specifically, we found in 45 MS patients that vitamin B12 levels were significantly lower in those who experienced the onset of first neurological symptoms prior to age 18 years (N = 10) compared to patients in whom the disease first manifested after age 18 (N = 35). In contrast, serum folate levels were unrelated to age of onset of the disease. As vitamin B12 levels were statistically unrelated to chronicity of illness, these findings suggest a specific association between the timing of onset of first neurological symptoms of MS and vitamin B12 metabolism. In addition, since vitamin B12 is required for the formation of myelin and for immune mechanisms, we propose that its deficiency in MS is of critical pathogenetic significance. PMID- 8407161 TI - The bioavailability of supplemental biotin in cattle. AB - A trial using 12 yearling heifers was carried out to test whether biotin metabolism and bioavailability are influenced by continuous dietary supplementation with biotin. Six of these heifers received no biotin supplementation (controls), while six received a daily dietary supplement of 20 mg biotin over the whole experimental period of four months. During each of three test periods (on days 14 and 21, 56 and 63, and 118 and 124), single test dosages of 40 mg (oral) and 5 mg (intravenous) biotin were given to each animal in a crossover test design. Blood samples were collected up to 72 h after each of these single doses, and at approximately two-weekly intervals for the assessment of baseline values. Serum biotin levels were determined by an ELSA test. Areas under the curves (AUC) were calculated as the target parameter for the assessment of the bioavailability of orally administered biotin. Serum biotin baseline levels were 300-800 ng/l in the controls and 3000-8000 ng/l in the supplemented animals. In both groups, AUC values in the first test period (days 14 and 21) were significantly higher than in subsequent periods. However, the biotin supplementation showed no significant effect. There was no significant difference in elimination half-lives between groups with and without biotin supplementation. The range was 5-18 h. Furthermore, there was no significant difference in the bioavailability of biotin between the test periods or between the biotin supplemented and unsupplemented animals. Overall bioavailability was 48%. PMID- 8407162 TI - Biochemical consequences of biotin deficiency in osteogenic disorder shionogi rats. AB - The biological consequences of biotin deficiency in rats were investigated using osteogenic disorder Shionogi rats which have a hereditary defect in ascorbic acid synthesizing ability. Decrease of liver ascorbic acid content and fasting plasma glucose and an increase of plasma non-esterified fatty acid (NEFA) appeared in biotin deficient rats fed a diet containing 200 mg ascorbic acid per 100 g diet, compared with the pair fed control. On the other hand, in the case of rats fed a diet containing 500 mg ascorbic acid, although the clinical features of biotin deficiency developed, the ascorbic acid contents of liver and adrenal gland increased in comparison with those of AsA 200 mg groups, and the alterations of plasma levels of glucose and NEFA were improved partially in glucose and greatly in NEFA, respectively. This suggests that ascorbic acid may be consumed in the improvement of the metabolic impairments induced by biotin deficiency. PMID- 8407163 TI - Changes in serum levels of vitamin B12, feed intake, liveweight and hematological parameters in cobalt deficient small east African goats. AB - Thirty short-horned East African goats were divided into two groups (treatments 1 and 2) consisting of fifteen goats each. All the goats were fed on a cobalt deficient diet containing 0.02 mg Co/kg of dry matter for a period of twenty three weeks. Goats in treatment 1 were supplemented with an oral cobalt chloride drench. Cobalt deficiency developed in goats in treatment 2 after ten weeks when their serum vitamin B12 concentration fell below 200-250 pg/ml. Between the third and the twenty-third weeks of the experiment, the mean serum concentration of vitamin B12 was significantly (P < 0.01) higher in cobalt supplemented (289.6 +/- 40.76 pg/ml) than deficient (142.8 +/- 28.27 pg/ml) goats. Total dry matter intake, intake per metabolic body weight and liveweight changes were not significantly (P > 0.05) different between goats in the two treatments. Although packed cell volume, hemoglobin concentration, and erythrocyte count were significantly (P < 0.05) higher in cobalt adequate than deficient goats, these blood parameters were within the normal range for goats. It is suggested that although serum vitamin B12 standard for sheep is applied for goats, the latter species is likely to be more resistant to low dietary cobalt intake than sheep. PMID- 8407164 TI - Tissue concentrations of water-soluble vitamins in normal and diabetic rats. AB - Changes in circulating and tissue concentrations of several vitamins have been reported in diabetic animals and human subjects. In this study, the effect of short-term (2 weeks) streptozotocin diabetes on folate, B6, B12, thiamin, nicotinate, pantothenate, riboflavin and biotin in liver, kidney, pancreas, heart, brain and skeletal muscle of rats was investigated. The tissue distribution of vitamins varied widely in normal rats. Diabetes significantly lowered folate in kidney, heart, brain, and muscle; B6 in brain; B12 in heart; thiamin in liver and heart; nicotinate in liver, kidney, heart and brain; pantothenate in all tissues; riboflavin in liver, kidney, heart, and muscle. These results indicate that experimental diabetes causes a depression of several water-soluble vitamins in various tissues of rats. PMID- 8407165 TI - Simultaneous determination of nicotinic acid and its two metabolites in human plasma using solid-phase extraction in combination with high performance liquid chromatography. AB - A high performance liquid chromatographic method for the simultaneous quantitative determination of nicotinic acid, nicotinamide and nicotinuric acid in human plasma is described. The method is based on solid phase extraction in combination with ion-paired reversed phase high performance liquid chromatography. Recoveries of nicotinic acid, nicotinamide and nicotinuric acid are 92.9, 95.8 and 87.8%, respectively. The procedure shown was applied to the determination of the plasma time-concentration profile of nicotinic acid, nicotinamide and nictotinuric acid after nicotinic acid ingestion in humans. PMID- 8407166 TI - Effect of diets high in sodium and potassium on the magnitude of theophylline induced hypercalciuria in the rat. AB - The effects of dietary sodium and potassium, given as salts of chloride or bicarbonate, on theophylline-induced hypercalciuria in the rat were investigated. In experiment 1, rats fed diets containing 0.3 mmol/g diet NaCl, KCl, NaHCO3, or KHCO3 exhibited a calciuria of NaCl > KCl > controls = NaHCO3 = KHCO3. This study indicated that cation effects were dependent on the accompanying anion, however the dose of added salt was inadequate to produce a consistent, significant salt effect. A second experiment was run in which animals were given salts at 0.4 mmol/g diet; the salt-induced pattern of calciuria was similar. After one week of salt pretreatment, rats were given diets containing theophylline (7.72 mumol/g diet), which induced calciuria all groups regardless of salt pretreatment. Urinary volume and urinary phosphate changes were parallel to each other and not to the changes in urinary calcium induced by salt or theophylline. Changes in urinary excretion of prostaglandin E2 were directly proportional to changes in urinary calcium. In the rat, pretreatment with sodium bicarbonate or sodium chloride reduces the extent of theophylline-induced diuresis, but does not reduce theophylline-induced hypercalciuria. Further, potassium chloride increases and potassium bicarbonate does not reduce urinary calcium excretion. Theophylline induced calciuria is correlated with prostaglandin E2 excretion rates. PMID- 8407167 TI - Effects of alpha-tocopherol on antioxidant enzyme activity in human fibroblast cultures. AB - The activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (Cat) were determined in human fibroblast cultures at four concentrations of exogenous alpha-tocopherol: 0.2, 2.5, 10 and 50 micrograms/ml of culture medium, or without alpha-tocopherol. Relationships between alpha tocopherol levels and the activities of SOD and GPx were identified. The cellular alpha-tocopherol level correlated with GPx activity (p < or = 0.01) and inversely correlated with SOD activity (p < or = 0.003), but only when alpha-tocopherol was added to the culture medium. The variations in the cellular GPx/SOD ratio depended on the level of cellular alpha-tocopherol (p < or = 0.001). Furthermore, there was a strong inverse correlation between SOD and GPx activity (p < or = 0.0001). Cat activity did not correlate either with cellular alpha-tocopherol concentration, or with SOD or GPx activity. These results underline the complex interplay between alpha-tocopherol and other antioxidant systems in human fibroblast cultures. PMID- 8407168 TI - Bovine plasma vitamin A responses to selected nutrients, monensin, and endophytic fescue. AB - Previous research at this station adapted a maximal dose response (MDR) method of evaluating vitamin A status and utilization for use in beef cattle. This method was used in two experiments. In this first experiment, forty-eight crossbred steers (average weight, 284 kg) were fed diets supplemented with salt, monensin or both, and injected with vitamin E, zinc or selenium. Steers receiving monensin had higher (38.5 micrograms/dl, monensin; 31.0 micrograms/dl, controls) initial plasma concentrations of vitamin A (P = .14). However, monensin did not affect post-dosing (MDR) vitamin A concentrations. None of the other dietary treatments or injections affected either pre- or post-dosing concentrations of vitamin A in the plasma. In a second experiment 23 lactating multiparous beef cows (average weight, 500 kg) grazing either fungal endophyte-infected or endophyte-free tall fescue were used to assess possible influences of infected fescue upon vitamin A metabolism. Fungal endophyte infection did not affect either pre- (44.9 micrograms/dl, end-noninf.; 47.7 micrograms/dl, end-inf.) or post-dosing (57.2 micrograms/dl, end-noninf.; 59.3 micrograms/dl, end-inf.) vitamin A concentrations. PMID- 8407169 TI - Rapid determination of blood serum retinol by reverse phase open column chromatography. AB - An aqueous, reverse phase, open column procedure (RP C18) was developed for the rapid determination of retinol in serum. The samples were extracted with hexane and separated on the column with methanol and water as the mobile phase. Two methods were developed depending on the size of the sample. For 2 ml of serum a spectrophotometer was used to quantify the sample and for 0.1 ml of serum a fluorometer was used. Phytofluene was retained on the top of the column and the column was reusable. The agreement between HPLC and the RP C18 open column was good. The lower limit of detection for micromethod was about 1.25 micrograms of retinol per dl of serum and the recovery of added retinol was quantitative. PMID- 8407170 TI - Plasma retinol and alpha-tocopherol concentrations in insulin-dependent diabetes mellitus: their relationship to microvascular complications. AB - Plasma alpha-tocopherol and retinol, both assayed by an HPLC method, have been evaluated in a group of 60 patients affected by insulin-dependent (type 1) diabetes mellitus, stratified according to the presence of retinopathy and nephropathy diagnosed by an urinary albumin excretion rate ranging between 20 and 200 micrograms/min (microalbuminaria) or > 200 micrograms/min (macroalbuminuria), all of whom were compared with 26 healthy controls strictly matched for age and sex. Plasma lipids and age were positively correlated with plasma retinol and alpha-tocopherol in both diabetic and control subjects. Either plasma retinol or its ratio to cholesterol were significantly and independently reduced in the younger subset of diabetics, as compared to controls, independently from other confounding variables, while plasma alpha-tocopherol was unchanged in diabetic subjects and in healthy controls. Retinopathy was not associated with altered levels of both plasma alpha-tocopherol or retinol. The presence of increased urinary albumin excretion was associated with higher plasma levels of alpha tocopherol and, only for macroalbuminuria, of retinol. However, after processing the data by a multivariate model, nephropathy was characterized by an increase only in plasma alpha-tocopherol. In conclusion, according to our findings, plasma retinol is significantly decreased in younger insulin-dependent diabetic patients while alpha-tocopherol is significantly altered in diabetic patients with nephropathy. PMID- 8407171 TI - Anticarcinogenic effect of common carotenoids. AB - Of the common carotenoids present in food, beta carotene, alpha carotene, lycopene, lutein, zeaxanthin as well as canthaxanthin can be considered potential prophylactic agents against carcinogenesis. They are absorbed by the human organism in reasonable amounts, and they have antioxidant properties, immunomodulating effects and may possibly influence gene expression enhancing gap junction communication. Recent suggestions that beta carotene may be metabolized directly to retinoic acid in retinoic acid target tissue and the discovery of retinoic acid nuclear receptors open up new perspectives for research. The best established chain of evidence for a protective effect of carotenoids against cancer development is available for beta carotene. Positive effects were observed in cell culture and experimental animal studies as well as in dietary and blood level studies in humans. More conclusive evidence will be provided by double blind intervention trials in humans that are in progress. Beta carotene appears to be active in the promotion phase of carcinogenesis stabilizing initiated cells. Canthaxanthin, which has often been included in animal experiments for comparative purposes having little or no provitamin A activity, also exhibits strong protective effects. Of the other carotenoids only limited data are available. Depending on the experimental model used, lycopene, lutein or alpha carotene was particularly active. In preliminary human blood level studies, lycopene was inversely associated with cancers of the pancreas and cervix. Much work remains to be done. Of particular interest is the question of organ specificity of individual carotenoids. PMID- 8407172 TI - Basal cell carcinoma of the eyelids. PMID- 8407173 TI - Cryotherapy for retinoblastoma. PMID- 8407174 TI - Plaque radiotherapy for retinoblastoma. PMID- 8407175 TI - Merkel cell carcinoma of the eyelids. PMID- 8407176 TI - Subclinical metastasis of uveal melanoma. PMID- 8407177 TI - Plaque radiotherapy for uveal melanoma. PMID- 8407178 TI - Local resection of posterior uveal tumors. PMID- 8407179 TI - Approach to counseling patients with posterior uveal melanoma. PMID- 8407180 TI - Tumors of the optic nerve head. PMID- 8407181 TI - Metastatic tumors to the uvea. PMID- 8407182 TI - Magnetic resonance imaging of orbital tumors. PMID- 8407183 TI - Biopsy techniques for orbital tumors. PMID- 8407184 TI - Lacrimal gland tumors. PMID- 8407185 TI - Metastatic tumors to the orbit. PMID- 8407186 TI - Squamous cell carcinoma of the conjunctiva. PMID- 8407187 TI - Rhabdomyosarcoma of the orbit. PMID- 8407188 TI - Immunohistochemical diagnosis of intraocular lesions. PMID- 8407189 TI - Malignant melanoma of the conjunctiva. PMID- 8407190 TI - Tumors of the caruncle. PMID- 8407191 TI - Magnetic resonance imaging of intraocular tumors. PMID- 8407192 TI - Genetic aspects of uveal melanoma. PMID- 8407193 TI - Sebaceous gland carcinoma of the eyelids. PMID- 8407194 TI - Clinical genetics of retinoblastoma. PMID- 8407195 TI - Fine-needle aspiration biopsy of suspected intraocular tumors. PMID- 8407196 TI - Hydroxyapatite orbital implant after enucleation for intraocular tumors. PMID- 8407197 TI - Photocoagulation of retinoblastoma. PMID- 8407198 TI - [The pathogenesis of rheumatic diseases. Role of infections]. PMID- 8407199 TI - [Imaging procedures in rheumatology]. PMID- 8407200 TI - [Therapy of rheumatoid arthritis at the turn of the century. Current status and trends of the '90's]. PMID- 8407201 TI - [New therapeutic possibilities. Future aspects]. PMID- 8407202 TI - [The role of surgical treatment of rheumatic diseases]. PMID- 8407203 TI - [29-year-old Senegal patient with acute thoracic discomfort and severe shoulder pain]. PMID- 8407204 TI - [63-year-old patient with cachexia in metastatic carcinoid]. PMID- 8407205 TI - [Characteristics of low-dose methotrexate therapy in rheumatoid diseases]. PMID- 8407206 TI - [Which courses of chronic replicating hepatitis C should be treated with interferon?]. PMID- 8407207 TI - [Infectious endocarditis]. PMID- 8407208 TI - Laminin receptors and laminin-binding proteins during tumor invasion and metastasis. AB - Interactions between cancer cells and laminin, a major component of basement membranes, play a critical role at several steps of the complex process of tumor invasion and metastasis. These interactions are mediated through a large variety of cell surface proteins designated as laminin receptors and/or laminin-binding proteins. The growing number of identified laminin binding proteins and domains of this glycoprotein that are biologically active illustrate the complexity of cellular interactions with laminin. The 67-kD laminin receptor (67 LR) was the first protein identified in 1983 as a high-affinity laminin receptor, and its expression is dramatically increased in a large variety of cancer cells. The 67 LR is the subject of several controversies and confusion generated mainly by two events. First, the identification of several new laminin-binding proteins has raised the difficult task of attributing specific functions to specific receptors in contrast to initial beliefs that all cellular laminin-driven biological activities were mediated through the 67 LR. The second source of controversy is the large molecular-weight discrepancy between the 37-kD polypeptide encoded by the 67 LR cDNA clone and the mature 67 LR. In this manuscript, a critical and extensive review of the data accumulated on the 67 LR is presented regarding both its molecular structure and its role during tumor invasion and metastasis. A hypothetical model of the 67 LR is also proposed. Since the first identification of the 67 LR, at least 14 other cell surface molecules have been reported to be potential laminin receptors or laminin-binding proteins. These include members of the beta-galactoside-binding lectin family, seven members of the integrin family, the galactosyltransferase and some not yet fully characterized cell surface molecules. These laminin receptors and laminin-binding proteins are also described and their functions are also discussed with a particular emphasis on their participation in the constitution of the invasive phenotype. PMID- 8407209 TI - Detection and localization of mRNAs encoding matrix metalloproteinases and their tissue inhibitor in human breast pathology. AB - Matrix metalloproteinases (MMPs) are a group of enzymes thought to be responsible for both normal connective-tissue-matrix remodelling and the accelerated breakdown associated with tumor development. These MMPs and tissue inhibitor of MMPs (TIMP1) could be expressed by either the cancer or the stromal cells. Expression of mRNAs encoding interstitial collagenase (MMP1), 72-kD type IV collagenase (MMP2) and stromelysin (MMP3), which are probably involved in tumor invasion and metastasis, and of TIMP1 were studied in human mammary pathology by in situ hybridization and Northern blot analysis. Out of 6 benign lesions, 2 expressed MMP2 mRNAs. mRNAs encoding MMP1 and MMP3 were detectable in occasional stromal and tumor cells in 2 out of 17 carcinomas. Thirteen out of 17 cancers expressed MMP2 mRNA throughout the tumor in stromal cells close to noninvasive tumor clusters and well-differentiated invasive cancer cells. TIMP1 mRNA expression was detected in noninvasive and well-differentiated invasive tumor cells. These data suggest that there is a cooperation between tumor and stromal cells, in particular for the production of 72-kD type IV collagenase, involved in the disruption of basement membranes. A lack of TIMP1 expression from invasive cancer cells would also contribute to matrix destruction. PMID- 8407210 TI - Characteristics of a metastatic variant to the liver of human rectal adenocarcinoma cell line RCM-1. AB - The interaction of human rectal adenocarcinoma cell line RCM-1 cells with extracellular matrix components, was studied to elucidate the key steps in the liver metastasis of colorectal carcinomas. Highly metastatic variant L-10 cells selected from the metastatic foci of the liver after intrasplenic implantation in nude mice and its parental L-0 cells were used. L-10 cells showed a greater ability to adhere to laminin, fibronectin, and type I and type IV collagens than did L-0 cells but less haptotactic activity than that of L-0 cells to type I or type IV collagen, possibly due to the formation of cellular aggregates. In vitro invasion activities of both cell lines to basement membrane components (Matrigel) or type I collagen were minimal but enhanced by the addition of 12-O tetradecanoylphorbol-13-acetate (TPA). L-10 cells showed greater ability to invade Matrigel than did L-0 cells, while L-0 cells exhibited higher activity in the invasion of type I collagen than did L-10 cells. TPA did not increase the production of metalloproteinases by both cells when analyzed by gelatin zymography. Based on the differences between the two cell lines, we postulated the following: (1) the high metastatic potential of L-10 cells was due to a greater capacity to attach to and cross the basement membrane; (2) TPA directly enhanced tumor cell invasiveness, not via the increased secretion of metalloproteinases; and (3) haptotactic migration had no significant correlation with the increased metastatic potential of L-10 cells. PMID- 8407211 TI - A modified neural network model of tumor cell interactions and subpopulation dynamics. AB - Tumors consist of phenotypically heterogeneous subpopulations of cells which are frequently affected by both autocrine and paracrine factors. Applying concepts from neural network theory, we have developed a computer model of chemical communication among hypothetical tumor cells, which simulates some of the complex epigenetic behavior of real tumors. Deletion of subpopulations often destabilized the whole population. The impact of deletion of specific subpopulations was affected by (a) which subpopulation was deleted, and (b) the timing of the deletion during tumor progression. PMID- 8407212 TI - S-cone function in patients with retinitis pigmentosa. AB - PURPOSE: To determine whether short-wavelength-sensitive (S-) cones are more severely damaged in patients with retinitis pigmentosa than long-wavelength sensitive (L-) and middle-wavelength-sensitive (M-) cones. To determine whether there are differences in the amount of S-cone damage in patients with dominant versus nondominant inheritance patterns. To accomplish these goals with methods that provide information not furnished by previous studies with two-color increment thresholds. METHODS: Acuity mediated by the S-cones was measured in 56 patients with retinitis pigmentosa, and the electroretinogram (ERG) generated by the S-cones was measured in 11 of these patients. Mixed L- and M-cone acuity, mixed L- and M-cone ERGs, and clinical full-field rod and cone ERGs were obtained for all patients. Data for both dominant and nondominant patient groups were compared with data from age-matched normal subjects. RESULTS: Only the nondominant group had reduced S-cone acuity, and 43% of patients in this group had selective reduction of S-cone acuity. In this particular sample the dominant and nondominant groups were comparable in clinical full-field ERG parameters and mixed L- and M-cone acuity, so the difference in S-cone acuities is not due to the dominant group having less advanced retinal degeneration. All 11 patients tested had reduced S-cone ERGs, 6 with significantly greater loss in the S-cone ERG than in the mixed L- and M-cone ERG. CONCLUSIONS: These data provide evidence that retinitis pigmentosa can produce greater loss of S-cones than L- and M cones, and that this selective loss is primarily seen in patients with nondominant forms of retinitis pigmentosa. PMID- 8407213 TI - Structural changes of the interphotoreceptor matrix in an inherited retinal degeneration: a lectin cytochemical study of progressive rod-cone degeneration. AB - PURPOSE: In the retinal disorder progressive rod-cone degeneration (prcd) in miniature poodle dogs, the photoreceptor layer degenerates slowly in the course of 5 to 7 years. Components of the interphotoreceptor matrix form a continuous extracellular lattice around photoreceptors. The purpose was to study the photoreceptor cell-matrix interactions during the disease and degeneration phases. Because degeneration rate was slower in cones, the authors also wanted to investigate whether there was a link between the degeneration and the photoreceptor-specific interphotoreceptor matrix domains. METHODS: Rod- and cone specific interphotoreceptor matrix domains were examined during two periods: before morphological signs of disease had appeared and during the degenerative stages. Two lectin probes were used; wheat germ agglutinin and peanut agglutinin. By their affinity for terminal carbohydrates, the lectins visually separated the two photoreceptor-specific domains and allowed follow-up of the fate of the rod and cone matrices separately. RESULTS: Before and during the course of disease, the lectin distribution in rod and cone domains remained normal, however, in the degenerative phase of the disease, there were structural changes in the matrix domains. The matrix connections between the individual domains was disrupted and single domains were formed. Cone domains and, to a lesser degree rod domains, were thickened around the inner and outer segments. CONCLUSIONS: The changes occurring in the photoreceptor-specific domains were indicative of structural adaptation to cell death and to degenerative conditions. There was no evidence of an active involvement of the interphotoreceptor matrix components studied in the disease process. PMID- 8407214 TI - Retinal pigment epithelial transplants and retinal function in RCS rats. AB - PURPOSE: To determine if retinal pigment epithelium (RPE) transplantation maintains visual function in Royal College of Surgeons (RCS) strain of rats. METHODS: Twelve RCS rats received RPE transplants at 16 to 20 days after birth. The retinas were studied electrophysiologically and histologically from 3 to 10 months after transplantation and compared with 11 RCS controls and 11 normal rats of comparable ages. A microelectrode was guided to the transplant site visible by its pigmentation in the albinotic RCS retina to detect responses. RESULTS: Spontaneous ganglion cell activity was present in all retinas. Ganglion cell responses to light were detected in 9 of the 12 transplant eyes but not in any of the 11 controls. 96, 44, 140 units were encountered and 30%, 0%, 97% were driven by light respectively in transplant, control, and normal retinas. In transplants 36%, 29%, and 28% were driven at 3 to 4, 6 to 7, and 10 months after transplantation, respectively. Intraretinal ERGs with both a- and b-waves were recorded in 5 of the 8 transplants studied. None of the RCS controls studied had an IERG. The average IERG was 2.5 microV (SD = 1.9) in transplants and 59 microV (SD = 19) in normal retinas. The electrode track was traced to the transplant site in six of the seven retinas that were responsive to light and examined histologically. CONCLUSION: RPE transplants to RCS rats maintain retinal function in the transplant site for long periods of time. PMID- 8407215 TI - Effect of deferoxamine on retinal lipid peroxidation in experimental uveitis. AB - PURPOSE: To examine the effect of deferoxamine, an effective iron chelator, on experimental autoimmune uveitis. Because deferoxamine has been shown to reduce iron-catalyzed hydroxyl radical generation, the in vivo effect was sought in the experimental autoimmune uveitis-mediated retinal lipid peroxidation, which is presumably induced by the inflammatory cell-derived oxygen radicals including hydroxyl radicals. METHODS: The experimental uveitis was induced in Lewis rats by retinal S-antigen. Deferoxamine infusion by osmotic pumps was started 2 days before the onset of the disease and was continued for 7 days. The extent of retinal lipid peroxidation was measured by the production of conjugated dienes, ketodienes, and thiobarbituric acid active substances. The inflammation associated free radical activity was measured by the luminol-amplified chemiluminescence. RESULTS: Levels of conjugated dienes, ketodienes, and thiobarbituric acid reactive substances were significantly decreased in the deferoxamine-treated animals. With Student's t test, the P values are < 0.025 for conjugated dienes between deferoxamine- and sham-treated animals; < 0.025 for ketodienes between deferoxamine- and sham-treated animals; and < 0.01 for thiobarbituric acid reactive substances between deferoxamine- and sham-treated animals. With in vitro addition of 10 mM deferoxamine, the free radical generation of inflamed retina was suppressed by nearly 40%. CONCLUSIONS: The administration of deferoxamine resulted in reduction of retinal lipid peroxidation. Because photoreceptors contain a high proportion of polyunsaturated fatty acids, deferoxamine, in turn, will act to ameliorate the experimental autoimmune uveitis-mediated retinal degeneration. PMID- 8407216 TI - Levodopa/carbidopa for childhood amblyopia. AB - PURPOSE: To evaluate the efficacy and tolerance of two low doses of levodopa/carbidopa (25/6.25 mg, 50/12.5 mg) and placebo (Tums) in 20 children with amblyopia between the ages of 4 and 14 years. METHODS: A double-masked placebo-controlled randomized 8-hour study was performed during which subjects received one of two doses of levodopa/carbidopa or placebo, combined with occlusion of the dominant eye. Visual acuity was measured at baseline and at 1 and 5 hours after capsule ingestion. Tolerance was assessed by questionnaire and physical examination. RESULTS: Visual acuity significantly improved by one line, from an overall average of 20/121 to 20/96, in the amblyopic eyes of both groups that received levodopa/carbidopa. Visual acuity did not significantly change in the placebo group. Tolerance was similar among all three groups. CONCLUSION: Average dose levels of 0.95/0.24 mg/kg and 1.94/0.49 mg/kg of levodopa/carbidopa were found to be well tolerated and efficacious at temporarily improving visual acuity in amblyopic eyes of children. PMID- 8407217 TI - Temporal modulation perimetry: the effects of aging and eccentricity on sensitivity in normals. AB - PURPOSE: Temporal modulation perimetry (TMP) is a new test procedure designed to measure sensitivity to sinusoidal flickering stimuli throughout the central 27 degrees visual field. The purpose of this investigation was to determine the effect of age and visual field eccentricity on temporal modulation sensitivity. METHOD: In its present form, TMP is used to determine modulation sensitivity for three temporal frequencies (2, 8, and 16 Hz) at 45 visual field locations. Both eyes of 43 normal observers between 20 and 75 years of age were examined. RESULTS: Our results indicate that sensitivity to all temporal frequencies tested showed a decline with age, particularly in the peripheral visual field. Furthermore, the age-related sensitivity loss was more pronounced for 16 Hz than for 2 or 8 Hz. CONCLUSIONS: These data demonstrate an age-related loss in temporal modulation sensitivity in the peripheral visual field. They also provide a baseline comparison standard for evaluation of clinical patient populations. PMID- 8407218 TI - Confocal imaging of the alpha 6 and beta 4 integrin subunits in the human cornea with aging. AB - PURPOSE: The purpose of this study was to examine changes in the distribution of integrin subunits, alpha 6 and alpha 4, in the normal human cornea with age. METHODS: Thirty normal corneas were examined and divided into three groups; corneas from children younger than 2 years, corneas from adults 29 to 70 years, and corneas from adults older than 70 years. The corneas were frozen and the sections were cut, double-stained with monoclonal antibodies to the integrin subunits, and visualized with Texas Red or fluorescein using confocal laser scanning microscopy. Computer imaging was conducted to determine differences. RESULTS: The alpha 6 subunit was generally localized along the basal and lateral surfaces of basal epithelial cells and projected into Bowman's membrane. The beta 4 subunit was only present along the basal surface. Overall, the major age related difference was the loss of continuous alpha 6 and beta 4 subunits along the basal surface of basal epithelial cells. When reconstructed images from corneas of individuals older than 70 years were optically sectioned en face, the alpha 6 subunit appeared discontinuous. If the same optical images were viewed from corneas of younger individuals, the staining was continuous. The number and distribution of hemidesmosomes along the basal lamina did not change with age in the corneas examined. CONCLUSIONS: Using computer imaging associated with confocal laser scanning microscopy, we have demonstrated that there is an age related change in the localization of the alpha 6 and beta 4 subunits. PMID- 8407219 TI - Visual attention problems as a predictor of vehicle crashes in older drivers. AB - PURPOSE: To identify visual factors that are significantly associated with increased vehicle crashes in older drivers. METHODS: Several aspects of vision and visual information processing were assessed in 294 drivers aged 55 to 90 years. The sample was stratified with respect to age and crash frequency during the 5-year period before the test date. Variables assessed included eye health status, visual sensory function, the size of the useful field of view, and cognitive status. Crash data were obtained from state records. RESULTS: The size of the useful field of view, a test of visual attention, had high sensitivity (89%) and specificity (81%) in predicting which older drivers had a history of crash problems. This level of predictability is unprecedented in research on crash risk in older drivers. Older adults with substantial shrinkage in the useful field of view were six times more likely to have incurred one or more crashes in the previous 5-year period. Eye health status, visual sensory function, cognitive status, and chronological age were significantly correlated with crashes, but were relatively poor at discriminating between crash-involved versus crash-free drivers. CONCLUSIONS: This study suggests that policies that restrict driving privileges based solely on age or on common stereotypes of age related declines in vision and cognition are scientifically unfounded. With the identification of a visual attention measure highly predictive of crash problems in the elderly, this study points to a way in which the suitability of licensure in the older adult population could be based on objective, performance-based criteria. PMID- 8407220 TI - A comparison of the laser flare cell meter and fluorophotometry in assessment of the blood-aqueous barrier. AB - PURPOSE: To evaluate and compare the use of the Kowa laser flare cell meter and intravenous anterior chamber fluorophotometry in assessment of the blood-aqueous barrier after cataract surgery. METHOD: Laser flare and cell measurements and fluorophotometry were performed at 1 and 3 months after surgery in 48 eyes of 44 patients admitted for routine cataract surgery. The fellow pseudophakic eyes of these patients were used as controls. RESULTS: The two techniques measure different parameters, but both methods are able to document the integrity or breakdown of the blood-aqueous barrier. However, the laser flare cell meter is more sensitive in quantifying subtle changes in barrier function to large molecules (proteins). Various methods of assessing anterior chamber fluorophotometry data were also compared. Measurement of a diffusion coefficient (requiring the measurement of plasma fluorescence) was not found to be more sensitive than other methods and did not alter the clinical significance of data obtained from the measurement of anterior chamber fluorescence alone. CONCLUSIONS: Both the laser flare cell meter and fluorophotometry provide a method for the assessment of the postoperative blood-aqueous barrier. However, the laser flare cell meter is rapid, noninvasive, and relatively easier to use. Therefore, for clinical use, it has great practical advantages over fluorophotometry. PMID- 8407221 TI - Aqueous humor flow and flare in patients with myotonic dystrophy. AB - PURPOSE: Myotonic dystrophy is an autosomal dominant form of muscular dystrophy associated with a mutation that affects a gene on chromosome 19. Extremely low intraocular pressure is one of a constellation of clinical signs that sometimes accompany this disorder. This study was performed to determine if the ocular hypotony can be explained by aqueous humor hyposecretion. METHODS: Seventeen persons with myotonic dystrophy and seventeen age-matched controls were studied. Intraocular pressure, light scattering (flare), fluorescein clearance at various times of day, and the response of the eye to the topical timolol were measured. RESULTS: Intraocular pressure was 40% lower in the persons with myotonic dystrophy (8.4 mm Hg vs 14.0 mm Hg) and flare was 50% higher (55.8 mg/dl albumin equivalent vs 37.1 mg/dl albumin equivalent). Fluorescein clearance in persons with myotonic dystrophy was indistinguishable from normal at all times of day. Myotonic eyes responded normally to topical timolol. The physiological data of this study are most consistent with the conclusion that the rate of aqueous humor flow is reduced approximately 9%, and the rate of inward leakage of light scattering proteins is increased approximately 37% in well-established myotonic dystrophy with ocular hypotony. CONCLUSIONS: The reduced aqueous humor flow alone is insufficient to explain the ocular hypotony. We hypothesize that the hypotony is due primarily to atrophy of the ciliary muscle that increases fluid exchange between the anterior chamber and the anterior uvea with consequent enhancement of uveoscleral outflow. PMID- 8407222 TI - Effects of experimentally induced ischemia on dopamine metabolism in rabbit retina. AB - PURPOSE: To determine the role of dopamine in experimentally induced retinal ischemia. Experiment 1 was designed to study the effects of ischemic levels on dopamine (DA) metabolism. Experiment 2 evaluated the effects of ischemic duration on DA metabolism. The effects of recirculation time after ischemia on DA metabolism were investigated in experiment 3. METHODS: Ischemia was produced by raising intraocular pressure (IOP) in rabbits. Three levels of ischemia were used -level A, level B, and level C--representing 50%, 75%, and 100%, respectively, of the IOP necessary to produce total ischemia. Retinal levels of DA and its main metabolites, homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC), were measured by high-performance liquid chromatography with electrochemical detection. RESULTS: Experiment 1 showed that at ischemic level B, DA contents were significantly reduced, but neither DOPAC nor HVA concentrations were altered. The reductions of retinal DA, DOPAC, and HVA concentrations were seen at level C without altering the ratio index. Level A ischemia did not alter DA metabolism. In experiment 2, significant reductions in DA, DOPAC, and HVA concentrations were found at both 30 and 60 minutes in the ischemic group, whereas the ratio DOPAC/DA and (HVA + DOPAC)/DA were significantly decreased only in the 60-minute ischemic group. Five-minute total ischemia did not alter DA metabolism. In experiment 3, concentrations of DA were still significantly decreased at 30 minutes of recirculation after ischemia, but DOPAC and HVA levels were back to normal. However, the ratios of DOPAC/DA, HVA/DA, and (HVA + DOPAC)/DA were significantly elevated. After 90 and 240 minutes of recirculation, retinal DA contents had returned to normal values, but DOPAC and HVA concentrations as well as all ratio indices of DA metabolism were still enhanced. CONCLUSION: Dopamine metabolism is altered during and after retinal ischemia. Dopamine may play a contributing role in ischemia. PMID- 8407223 TI - Mast cells in human optic nerve. AB - PURPOSE: Mast cells are classically found in ocular tissues within the conjunctiva, choroid, and iris. The aim of this study was to examine their distribution in the optic nerve and its meninges. METHODS: Sixty-six human optic nerves were studied from normal subjects at autopsy, fetuses aborted for chromosomal abnormalities, and from enucleation specimens of patients with a variety of inflammatory, traumatic, neoplastic, and vascular disorders. Mast cells were identified using a stain for the enzyme chloroacetate esterase, and confirmed using toluidine blue, revealing metachromatic cytoplasmic granules. RESULTS: Mast cells were found scattered in the meninges of almost all optic nerves examined, frequently in perivascular locations, with densities up to 2325 mast cells/mm3 (mean 269.7 +/- 64.1 cells/mm3 in normal nerves). Mast cells were found in the optic nerve parenchyma in nerves from four eyes that had severe abnormality, often associated with neovascularization. Normal nerves, as well as nerves from fetuses aborted for congenital defects, had significantly fewer meningeal mast cells than those from eyes with inflammatory or vascular diseases. Degranulation of mast cells was observed more often in eyes with recent severe trauma. CONCLUSIONS: Based on their work and the work of others suggesting an association between mast cells and nervous system autoimmune disorders, the authors hypothesize a role for optic nerve mast cells in certain ocular inflammatory conditions. PMID- 8407224 TI - Expression of glial fibrillary acidic protein in rabbit Muller cells after lensectomy-vitrectomy. AB - PURPOSE: To assess retinal response after lensectomy-vitrectomy, the authors analyzed glial fibrillary acidic protein (GFAP), which retinal glial cells express in several pathologic conditions. METHODS: GFAP expression was examined by means of immunohistochemistry, Western blot analysis, and enzyme-linked immunosorbent assay (ELISA) in rabbit retina after lensectomy-vitrectomy. RESULTS: Muller cells had detectable GFAP immunoreactivity in peripheral retina 3 days after surgery. GFAP immunoreactivity was present in both peripheral and posterior regions 7 days, 14 days, and 6 months after surgery. The authors confirmed that anti-GFAP antibody recognized the 51 kDa protein specifically by Western blot analysis. Time-dependent increase of GFAP in peripheral retina was also obtained by ELISA: CONCLUSIONS: Such progressive GFAP accumulation in Muller cells, which are sensitive to microenvironmental changes, may reflect some underlying retinal response after lensectomy-vitrectomy. PMID- 8407225 TI - Iron-induced accumulation of lipofuscin-like fluorescent pigment in the retinal pigment epithelium. AB - PURPOSE: One of the most prominent changes that occurs in the retinal pigment epithelium during senescence is the progressive accumulation of the autofluorescent pigment lipofuscin. Experiments were conducted to evaluate the role of nonenzymatic oxidation of photoreceptor outer segments in retinal pigment epithelium lipofuscin formation. METHODS: Albino Fischer rats were given intravitreal injections of ferrous sulfate, a catalyst that promotes nonenzymatic lipid oxidation. At 2 hours, 24 hours, and 7 days after ferrous sulfate administration, the retinas were examined with fluorescence microscopy to assess the formation of fluorescent products. At these same time intervals, organic solvent extracts of the retinas and retinal pigment epithelium-choroid complexes were prepared. The extracts were analyzed with thin layer chromatography to assay for the presence of soluble fluorophores. The ultrastructural appearances of the retinas were examined at the same time points. RESULTS: At both 2 hours and 24 hours after the ferrous sulfate treatment, the photoreceptor outer segments displayed a yellow-green fluorescence emission that was not present in untreated eyes. Associated with this in situ fluorescence were a number of blue-green emitting fluorophores in organic solvent extracts that did not correspond to any of the fluorophores extracted from the retinal pigment epithelium of old animals. One week after the ferrous sulfate treatment, the photoreceptor cells had degenerated and the retinal pigment epithelium contained large amounts of an autofluorescent pigment with a golden-yellow emission typical of lipofuscin. The iron-induced fluorophores could not be extracted from this pigment into either chloroform or dichloromethane. CONCLUSIONS: The initial fluorophores that were formed as a result of nonenzymatic oxidation of outer segment components did not appear to be the same as those responsible for retinal pigment epithelium lipofuscin fluorescence. However, after the oxidized outer segments were phagocytosed by the retinal pigment epithelium, the latter cells became filled with a yellow-emitting fluorescent pigment that was similar in its fluorescence properties to lipofuscin. These observations suggest that lipofuscin fluorophores are not direct products of nonenzymatic lipid oxidation. However, some of these oxidation products may be modified after uptake by the retinal pigment epithelium to form insoluble lipofuscin fluorophores. PMID- 8407226 TI - Aldose reductase the major protein associated with naphthalene dihydrodiol dehydrogenase activity in rat lens. AB - PURPOSE: Recent studies have indicated that certain aldose reductase inhibitors prevent the formation of cataracts in naphthalene-fed rats. The study was designed to investigate whether aldose reductase itself is involved in the metabolism of naphthalene in the lens and the mechanism of this cataract formation. METHODS: Aldose reductase was purified from whole rat lens using a series of chromatographic steps that include gel filtration, affinity chromatography, and chromatofocusing. The dehydrogenase activity of the purified enzyme was evaluated with 1,2-dihydro-1,2-dihydroxynaphthalene (naphthalene dihydrodiol) as substrate. The same dehydrogenase activity was also examined with the recombinant protein obtained from rat lens aldose reductase clone. RESULTS: Throughout the purification steps, dehydrogenase activity with naphthalene dihydrodiol as substrate coeluted with aldose reductase activity assayed with DL glyceraldehyde as substrate. The purified aldose reductase, which appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, displayed dehydrogenase activity with naphthalene dihydrodiol as a substrate similar to that observed with the crude extract from whole rat lens. Recombinant protein from rat lens aldose reductase clone also displayed dehydrogenase activity similar to that observed with purified rat lens aldose reductase. Both the reductase and dehydrogenase activities of purified aldose reductase were inhibited by aldose reductase inhibitors. However, inhibition of dehydrogenase activity was less than reductase activity. Aldehyde reductase, an another nicotinamide adenine dinucleotide phosphate-dependent reductase, also displayed dihydrodiol dehydrogenase activity with naphthalene dihydrodiol and this activity was also inhibited by aldose reductase inhibitors. CONCLUSIONS: Aldose reductase displays dehydrogenase activity in addition to reductase activity. In rat lens aldose reductase is a major protein associated with naphthalene dihydrodiol dehydrogenase activity. This suggests that aldose reductase is linked to 1,2 dihydroxynaphthalene formation in rat lens and the subsequent formation of cataracts in naphthalene-fed rats. PMID- 8407227 TI - Cytokines cause cultured retinal pigment epithelial cells to secrete metalloproteinases and to contract collagen gels. AB - PURPOSE: Because retinal pigment epithelial cells in epiretinal membranes remodel and contract their surrounding extracellular matrix, investigations were performed to determine if these cells can produce matrix metalloproteinases and contract collagen gels in vitro in the presence of serum or cytokines. METHODS: Cells were grown on collagen gels and their production of metalloproteinases was measured using zymography. RESULTS: Cells grown in a three-dimensional collagen gel culture system produce several latent metalloproteinases that are secreted into the gel and the surrounding medium. These include molecules of 49, 56, 66, and 100 kD. In addition, an enzyme that is probably the active form of the 66 kD enzyme is present. When interleukin 1 beta is added to the cultures, latent 49 kD and 100 kD gelatinase production is greatly stimulated and an active form of both enzymes is also observed in the medium. In contrast, transforming growth factor beta has no stimulatory effect. The cells contract the collagen gel but this is small without cytokines; however, contraction is greatly enhanced in the presence of serum or interleukin 1 beta plus transforming growth factor beta. Contraction is unlikely to be the result of metalloproteinase action on the underlying extracellular matrix because complete inhibition of these enzymes has little effect. CONCLUSIONS: These results show that cytokines can cause cultured retinal pigment epithelial cells to produce metalloproteinases that can, when activated, degrade most collagens and other structural molecules in extracellular matrix. In addition, they can stimulate the contraction of extracellular matrix constituents but there is not a simple causal relationship between matrix remodeling and contraction. PMID- 8407228 TI - Lipids of human retina, retinal pigment epithelium, and Bruch's membrane/choroid: comparison of macular and peripheral regions. AB - PURPOSE: To understand the difference between macular and peripheral regions, tissue samples of human retina, retinal pigment epithelium, and Bruch's membrane/choroid were dissected and analyzed for lipid composition. METHOD: To facilitate dissections and enhance the recovery of tissues, eyecups were prefixed for 1 hour in 10% formalin (pH 7). Lipids were extracted from 4-mm trephined punches of tissues and analyzed by high-performance liquid chromatography. After separation of neutral lipids and phospholipids, total fatty acids in both lipid classes were quantitated. RESULTS: The major phospholipid classes in retina, retinal pigment epithelium, and Bruch's membrane/choroid were phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl inositol, and phosphatidyl serine; the major fatty acids were palmitic (16:0), stearic (18:0), and oleic (18:1). Although the three tissues had similar total fatty acid and phospholipid components, their relative compositions were different. Neutral lipid/phospholipid ratios in retinal pigment epithelium and Bruch's membrane/choroid were almost three times higher than in the retina. CONCLUSIONS: This study provides information about the lipid composition of macular and peripheral regions of the human retina, retinal pigment epithelium, and Bruch's membrane/choroid. The methodology employed enabled study of lipids in small amounts of tissue, which will be of value in investigating the biochemical aspects of age-related macular degeneration. PMID- 8407229 TI - Nonocular Chlamydia infection and risk of ocular reinfection after mass treatment in a trachoma hyperendemic area. AB - PURPOSE: The presence of nasal discharge on a child's face increases the risk of active trachoma, suggesting that Chlamydia trachomatis in nasal secretions may be a possible source of ocular reinfection. The prevalence of chlamydia in nasal secretions and the risk of reinfection after mass treatment was investigated in a hyperendemic area of Tanzania. METHODS: In one village a total of 232 children aged 1 to 7 years were followed before and after mass treatment. Clinical trachoma, and microbiologic evidence of chlamydia, were assessed at baseline, 2 and 4 weeks into mass treatment, and 4 weeks after treatment stopped. The presence of chlamydia in ocular and nasal secretions was determined by polymerase chain reaction-enzyme immunoassay techniques. RESULTS: Of the 232 children, 59% had clinical trachoma and 27% had nasal specimens positive for chlamydia. Children with positive ocular chlamydia specimens and/or clinical trachoma were significantly more likely to have positive nasal specimens. At the end of mass treatment, only 4% of children had positive ocular specimens. However, 1 month after treatment stopped, the incidence of new infection was 21%. The rate of new ocular infections in those who had negative ocular specimens after treatment was similar between those who had positive and those who had negative nasal specimens at baseline. Positive ocular specimens at baseline was not a predictor of risk of new infection after treatment (odds ratio = 1.18, 95% confidence interval = 0.58, 2.40), suggesting these new infections were not the result of latent or persistent organism. CONCLUSIONS: These data do not support a role for nasal secretions in causing reinfection after treatment. One mass topical treatment alone is unlikely to be effective in trachoma hyperendemic areas as shown by the rapid re-emergence of infection. PMID- 8407230 TI - Videographic Hirschberg measurement of simulated strabismic deviations. AB - PURPOSE: To demonstrate the potential use of subpixel image processing methods to perform automated Hirschberg measurements of strabismic deviations using relatively inexpensive personal computer hardware; to determine if the method might allow screening for strabismus using full-face video images obtained from a distance of 1 meter. METHODS: Strabismic deviations (< 25 prism diopters) were simulated by means of induced asymmetric fixation. A ring of coaxial infrared light-emitting diodes (LED) were used to generate first Purkinje reflexes. Computerized image analysis with subpixel processing was used to measure the locations of the first Purkinje reflexes and pupil centers of video images of 10 normal subjects, following the technique of the clinical Hirschberg test. The apparent strabismic deviation was calculated from the relative asymmetry of the center of the corneal reflex ring to the pupil center in each eye. RESULTS: In 10 normal subjects, there was a statistically significant linear correlation of Hirschberg horizontal reflex deviation with asymmetric fixation pseudo-esotropia (0.85 > or = r2 > or = 0.99, P < 0.05). CONCLUSIONS: The Hirschberg test is used manually to detect strabismus in infants and children but requires a highly skilled examiner. The image processing method described here requires no operator interpretation and may make the test more applicable. The results suggest that this technology may be appropriate for a screening instrument. PMID- 8407231 TI - Health hucksters continue preying on Iowans. PMID- 8407232 TI - Vulnerability leads patients to non-proven treatments. PMID- 8407233 TI - Unusual case. PMID- 8407234 TI - Developments in radiology: techniques new, refined and revisited. PMID- 8407235 TI - Remediation through personalized CME (Part II). PMID- 8407236 TI - Possible role of human foamy virus in Graves' disease. AB - Human foamy virus (HFV) is a member of the retroviral family of Spumaretrovirinae. In addition to the three retroviral structural genes, gag, pol and env, HFV also contains regulatory sequences, called bel. Foamy viruses have been previously associated with human thyroid disease, notably DeQuervain's thyroiditis. In indirect immunofluorescence tests we have demonstrated the reactivity of the thyroid glands of 7/7 patients with Graves' disease and antibodies to HFV gag proteins. No reactivity was observed with antibodies to pol, env and bel proteins. Nine thyroids of patients with struma parenchymatosa, 4 with follicular carcinoma and 2 normal thyroids were negative throughout. From the thyroids of 5 patients with Hashimoto's disease, 4 were negative and 1 showed a single small focus of anti-gag antibody reactivity. The uniform immunofluorescent staining was restricted to the basal and lateral intercellular areas between the thyroid epithelial cells. Extension of these studies to the retrobulbar tissue of 1 Graves' disease patient with malignant exophthalmus revealed positive staining with anti-gag antibodies of fibroblasts and fat cells but not eye muscles. Furthermore, we were successful in establishing several T cell lines derived from the retrobulbar tissue of this patient. They were CD8+ and proliferated, in contrast to peripheral blood cells, upon cocultivation with autologous retroorbital fibroblasts. It remains to be determined whether these observations are of relevance in the pathogenesis of Graves' disease. PMID- 8407237 TI - Viruses and autoimmune liver disease. AB - Autoimmune hepatitis is characterized by hypergammaglobulinemia, female predominance, autoantibodies and a good response to immunosuppression, and is based on specific autoantibodies and clinical characteristics. Several subgroups may be distinguished. As in most autoimmune diseases, the etiology is unknown. The association of autoimmune hepatitis with a viral etiology is most prominent in autoimmune hepatitis type 2 which is characterized by liver/kidney microsomal (LKM-1) autoantibodies against cytochrome P-450 II D6. Depending on the geographical origin of the patients, a specific proportion of patients with autoimmune hepatitis type 2 is associated with hepatitis C virus infection. These HCV RNA-positive patients are older, female predominance is not profound, and response to immunosuppression is generally low compared to the HCV-negative patients with autoimmune hepatitis type 2. The genetic background is unclear. HCV sequence analysis revealed that HCV genotype II is prominent in HCV-positive autoimmune hepatitis type 2. HCV mutants with deletions in the HCV envelope region were identified. The relevance of these HCV mutants for the induction of autoimmunity needs to be characterized further. The HCV-negative population of patients with autoimmune hepatitis type 2 seems to have a relation with herpes simplex virus (HSV-1) infection since the B-cell epitope of cytochrome P-450 II D6, the major LKM-1 antigen, shares sequence homology with the IE-175 protein of HSV-1. In the HCV-negative population of autoimmune hepatitis type 2, HLA-DR3 and C4-AQ0 alleles are significantly increased. PMID- 8407238 TI - Mechanism and consequence of viral persistence in cells of the immune system and neurons. AB - Viral persistence depends on a virus having a non-lytic strategy of replication and the ability to escape immune surveillance. Cells of the immune system (lymphocytes/monocytes/macrophages) and central nervous system (neurons) are most often infected by DNA and RNA viruses that persist. Cytotoxic T lymphocytes (CTL) are the primary host defense that aborts or prevents viral persistence. Viral interaction with these specialized cells and of such infected cells with CTL is explored in this paper. PMID- 8407239 TI - Virus- and immune-mediated liver damage in hepatitis. AB - It has been assumed for many years that liver damage associated with hepatitis B virus (HBV) infection is due to cytolytic immune reactions directed at viral antigens expressed on infected liver cells. Recent studies, however, suggest that in some clinical settings, changes in the HBV genome, possibly selected by immune pressure, can interfere with the export of viral proteins from hepatocytes and lead to direct virus-induced liver damage. PMID- 8407240 TI - Hepatitis A: hepatotropism and influence on myelopoiesis. AB - Immunopathologic mechanisms leading to liver tissue injury in hepatitis caused by hepatitis A virus (HAV) were studied in an autologous in vitro model. Data show virus-specific killing by liver-infiltrating T lymphocytes in man and support the hypothesis that hepatocellular damage as well as efficient elimination of virus infected hepatocytes is mediated by HLA-restricted, HAV-specific CD8+ T lymphocytes. Furthermore, experimental results demonstrate that human interferon gamma produced by HAV-specific T cells may act as a key factor in T-cell-promoted clearance of HAV-infected hepatocytes. Besides the well-known hepatotropism, the myelotropic properties of HAV have some important clinical implications. Perturbations of hematopoietic regulation, ranging from transient granulocytopenia to rare cases of bone marrow failure, are associated with HAV infection. In an attempt to elucidate the pathogenetic mechanisms, we could show a direct suppressive effect of HAV on human bone marrow progenitors and a significant progressive decline in these cells in HAV-infected long-term bone marrow cultures. PMID- 8407241 TI - Molecular pathogenesis of enterovirus-induced myocarditis: virus persistence and chronic inflammation. AB - In situ hybridization studies have proved that myocardial enterovirus infections are detectable in all stages of acute and chronic enterovirus-induced myocarditis as well as in some patients with end-stage dilated cardiomyopathy, suggesting the possibility of myocardial enterovirus persistence. Possible enterovirus persistence in the human heart is supported by the discovery of enterovirus persistence in different murine models of chronic myocarditis, demonstrating that coxsackievirus B3, typically a cytolytic enterovirus, is capable of evading immunological surveillance in a host-dependent fashion. Progress is currently being made in unraveling the molecular mechanisms of enterovirus persistence, the diversity of host and virus genetics and their impact on the nature and severity of the disease. Apart from providing an etiologic diagnosis, there are therapeutic implications from the in situ demonstration of myocardial enterovirus infection. Evaluation of specific antiviral agents, for example interferons, may lead to the development of new therapeutic strategies capable of providing protection against myocardial enterovirus infection. PMID- 8407242 TI - HSV-1 neuroinvasiveness. AB - Neuroinvasiveness, the capacity of virus to enter and progress through the nervous system, may be accomplished by hematogenous or neural routes. We have been interested in defining HSV-1 genes specifically concerned with the neural pathway. In one system (involving HSV strain Ang), we have found that a single amino acid change in glycoprotein D, a viral membrane protein, confers invasiveness upon a noninvasive agent. Preliminary studies of another noninvasive agent, KOS, suggests that at least 2 genes are related to the phenotype. Experiments which establish specificity for the noninvasive phenotype and a discussion of the potential mechanisms involved in the glycoprotein D localization are also discussed. PMID- 8407243 TI - Influenza--a model of an emerging virus disease. AB - Influenza A viruses continue to emerge from the aquatic avian reservoir and cause pandemics. Phylogenetic analysis of the nucleotide sequence of all eight influenza A virus RNA segments indicate that all of the influenza viruses in mammalian hosts originate from the avian gene pool. In contrast to the rapid progressive changes in both the nucleotide and amino acid sequences of mammalian virus gene lineages, avian virus genes show far less variation and, in most cases, appear to be in evolutionary stasis. There are periodic exchanges of influenza virus genes or whole viruses between species giving rise to pandemics of diseases in humans, lower animals and birds. The periodic emergence of influenza viruses in mammalian species has been illustrated by the appearance of a new influenza virus in horses in northern China in 1989. Phylogenetic analysis of classical H1N1, avian-like H1N1 and human H3N2 viruses circulating in Italian pigs reveals that genetic reassortment is taking place between avian- and human like viruses in the European pig population. These studies provide evidence supporting the possibility that pigs serve as a mixing vessel for reassortment between influenza viruses in mammalian and avian hosts and raise the question of whether the next pandemic of influenza will emerge in Europe! PMID- 8407244 TI - Role of the PrP gene in transmissible spongiform encephalopathies. AB - The transmissible agent that causes spongiform encephalopathies such as scrapie, the prion, is believed to be devoid of nucleic acid and identical with PrPSc, a modified form of PrPC. PrPC is a normal host protein encoded by a single copy gene (Prn-p) and is found predominantly on the surface of neurons. PrPSc, in contrast to PrPC, is resistant to protease and accumulates intracellularly. Prusiner proposed that PrPSc, when introduced into a normal host, causes the conversion of PrPC or its precursor into PrPSc ('protein only' hypothesis). If indeed PrP is an essential component of the prion, then an animal devoid of the PrP protein should be resistant to scrapie. We generated homozygous PrP 'knockout' mice. These Prn-p0/0 mice showed no gross abnormalities, and microscopic examination of brain sections of normal and Prn-p0/0 mice revealed no differences. Prn-p0/0, Prn-p+/+ and Prn-p0/+ mice were inoculated with scrapie agent; the clinical response as well as the prion titer at different time points are being determined. PMID- 8407245 TI - Autoimmune diseases in humans, e.g. autoimmune rheumatic diseases. AB - In spite of increasing evidence that viruses and especially retroviruses could act as etiologic factors in autoimmune and especially autoimmune rheumatic diseases, clear-cut evidence for an involvement of these agents is still missing. Findings, which, for example, indirectly support the hypothesis that retroviruses might play a part, are the demonstration of antibodies to the gp24 in SLE and Sjogren's patients as well as the description of retroviral antigens in the inflamed synovium of rheumatoid arthritis patients. Furthermore, evidence comes from animal models that viruses, such as the Visna or Caprine arthritis encephalitis virus, induced chronic inflammatory diseases in sheep and goats. More recently, a mouse model for rheumatoid arthritis and Sjogren's syndrome was reported in mice transgenic for HTLV-1tax. It is hoped that, especially from the experimental animal models, the possible role of retroviruses as etiological factors in autoimmune rheumatic diseases can be clarified. PMID- 8407246 TI - Foamy viruses. AB - Foamy viruses share complex genome organization with lentiviruses and certain oncoviruses. The open reading frame 3' of env encodes a transcriptional transactivator. Distinct responsive sequences were identified in the long terminal repeats (LTRs) of simian (SFV-1 and SFV-3) and human foamy viruses (HFV). Transactivation of heterologous LTRs was described including those of simian and human immunodeficiency viruses. Foamy viruses persist for the whole lifetime in infected hosts (primates, cats, hamsters, cattle, and probably other mammals). The virus may be orally shed and transmitted, while being latent in various internal organs. Selective viral gene expression in the brains of mice transgenic for HFV has suggested a particular relationship to neural tissue. In latently SFV-3-infected cultured cells, methylation of proviral DNA is apparently involved in the control of latency. Demethylation as well as transfection with the transactivator were shown to be instrumental in viral reactivation. Natural infections with foamy viruses are common, elicit strong immune responses, and seem to be asymptomatic in nonhuman primates. Detection of such infections, however, may not be a triviality in man. While accidental transmission of foamy viruses to man is well documented, reported seroprevalence in human populations and the association of HFV with specific pathology (e.g. thyroiditis de Quervain, amyotrophic lateral sclerosis, and Graves' disease) are controversial and remain to be proven. PMID- 8407247 TI - Bovine spongiform encephalopathy: an appraisal of the current epidemic in the United Kingdom. AB - Bovine spongiform encephalopathy (BSE) is a food-borne infection of cattle caused by the use of contaminated meat and bone meal in concentrated feeds. The UK epidemic was initiated by a sudden exposure to infection in 1981-1982, which was associated with a dramatic reduction in the use of organic solvents in the manufacture of meat and bone meal. This change almost certainly removed two partial disinfection steps and allowed enough contamination with a scrapie-like agent to infect cattle. Although it is assumed that the epidemic originated with scrapie infection crossing the species barrier, cattle-to-cattle recycling of infection, via feed, amplified the epidemic very considerably. There would have been a strong tendency for recycling to select a single cattle-adapted strain of agent, and this strain of BSE could well be different from scrapie. There is evidence to support both predictions. Because the median incubation period of BSE is 4-6 years, clinical cases did not appear until 1985-1986, by which time the recycling of infection in cattle was probably well established. However, the average food-borne exposure to infection has remained low resulting in a mainly sporadic occurrence of BSE. Signs of an imminent decline in the epidemic were unmistakable early in 1993, which is over 4 years after the feeding of ruminant derived protein was banned to prevent new infections of cattle. PMID- 8407248 TI - Epstein-Barr virus and its interaction with the host. AB - Epstein-Barr virus (EBV) as a member of the herpesvirus family persists lifelong in the human body and causes diseases associated with virus replication (infectious mononucleosis, oral hairy leukoplakia) as well as neoplastic conditions such as nasopharyngeal carcinoma, B-cell lymphoma, Hodgkin's disease associated with viral latency. This complex biology relates to a highly regulated control of the persisting virus. Still, EBV is lytically produced in certain compartments of the human body. Epithelial cells were found to be of key importance for this. Various routes (cell fusion, IgA receptor-mediated uptake) were described for EBV to enter epithelial cells in the absence of CR2 receptor. Viral entry into cells, however, via CR2 receptor fusion or IgA mediated was not found to be sufficient for viral production. The molecular mechanisms for the lack of viral production in most target cells are primarily the presence of silencer activities and the early elimination of cells entering the lytic cycle. Only terminally differentiated epithelial cells are capable of supporting an efficient lytic cycle of EBV replication. EBV-mediated suppression of apoptosis as well as down-regulation of cellular and viral gene products, such as HLA molecules, which mediate recognition by the immune system, are important contributing factors to the development of these neoplasias where viral genes, possibly via interaction with anti-oncogenes, such as p53, in context with genetic and environmental factors play a key role. Novel diagnostic tools and a vaccine have been developed which could help to control EBV-related diseases. PMID- 8407249 TI - Hepatitis B virus: significance of naturally occurring mutants. AB - Mutations of viral genomes are normal biological events and result in the coexistence of viral genotypes in infected individuals ('quasispecies'). Hepatitis B virus (HBV) and the related animal hepadnaviruses have a mutation rate which is intermediate between DNA and RNA viruses because they replicate asymmetrically via reverse transcription of an RNA intermediate. HBV mutants affecting all known reading frames of the viral genome have been demonstrated in patients with acute fulminant or chronic HBV infection. Some of the mutations identified to date suggest a contribution to viral latency, low level HBV infection, the severity of liver disease and vaccine escape. Since most viral genomes carry more than one mutation and most individuals are infected by more than one variant, the demonstration of a causal relationship between a single mutation and a biological or pathobiological effect requires the in vitro and in vivo analysis of genetically defined mutants. Such analyses should allow a molecular understanding of the genetic contribution of HBV to the variable natural course of HBV infection, ranging from an asymptomatic healthy carrier state to acute or even fulminant hepatitis, chronic liver disease, liver cirrhosis and hepatocellular carcinoma. PMID- 8407250 TI - Experimental and spontaneous mucosal disease of cattle: a validation of Koch's postulates in the definition of pathogenesis. AB - The complex etiology of mucosal disease of cattle has been unravelled by experimental reproduction of the fatal syndrome, thereby validating Koch's postulates. The causal virus, bovine virus diarrhea virus (BVDV), has two biotypes, noncytopathogenic and cytopathogenic; the former can infect the early bovine fetus, induce immunotolerance and a lifelong persistent viremia (pv), whereas the latter biotype can superinfect the pv cattle and induce mucosal disease. The cytopathogenic biotype appears able to originate de novo from the persisting virus by molecular rearrangement (e.g. template switching) and has been shown to arise in a case of spontaneous mucosal disease. The animal had been maintained in isolation for an extended period. PMID- 8407251 TI - Diagnostics of persistent viruses: human cytomegalovirus as an example. AB - Infections with persistent viruses such as herpesviruses have become of significant clinical importance with the increasing number of immunocompromised patients at risk to suffer from severe disease. As antiviral chemotherapy is available for herpesvirus infections, the diagnostic methods for rapid and sensitive detection of symptomatic infection have been developed and recently refined. In human cytomegalovirus (HCMV), the use of recombinant viral antigens provides a rationale to improve serological assays. This may be of use for the discrimination of primary versus secondary infection. Early diagnosis of symptomatic HCMV infection in immunosuppressed patients can be most effectively achieved by the detection of a viral tegument protein, pp65, in peripheral blood leukocytes. This early diagnosis has been shown to be of major importance for the effective treatment of these patients. HCMV infection in solid organs can be demonstrated by immunohistochemistry using monoclonal antibodies against viral proteins. HCMV involvement in diseases of the central nervous system in AIDS patients can be verified by the detection of very small amounts of HCMV DNA in cerebrospinal fluid by polymerase chain reaction. This method may prove useful for monitoring HCMV encephalitis and neuropathy. PMID- 8407252 TI - Autoantibodies in viral hepatitis-related hepatocellular carcinoma. AB - Patients with hepatocellular carcinoma (HCC) develop autoantibodies to nuclear and nucleolar antigens (ANAs) which can be readily detected by immunofluorescence on cell substrates. The frequency of ANAs in HCC is 31% (57/184). The identity of three autoantigens was established as: NOR-90, nucleolus organizer region (doublet) polypeptides involved in RNA polymerase I transcription; fibrillarin, a component of nucleolar U3 RNP involved in pre-ribosomal RNA processing, and nucleophosmin/protein B23, a nucleolar protein involved in ribosome maturation and cell proliferation. Changes in ANAs were observed in some patients during transition from chronic liver disease to HCC and were manifested as seroconversion from ANA-negative to ANA-positive status by an increase in titers and changes in ANA specificities. Serum from a patient during this transition period was used to isolate a cDNA clone encoding a novel nuclear protein with structural motifs characteristic of a family of splicing factors. These observations support the notion that ANA responses in HCC might be driven by intracellular events related to transformation from the stage of chronic injury to the stage of malignancy. Changes in ANA profiles which were observed to precede clinically diagnosed HCC in some patients might be early markers of transformation. PMID- 8407253 TI - Measles virus-induced autoimmune reactions against brain antigen. AB - Recovery from viral infection is a result of very complex interactions between specific and nonspecific immune reactions and the infectious agent. A variety of immune mechanisms are undoubtedly important factors in this event and operate together in overcoming the infectious process. However, despite much available information about these viral defense mechanisms, it has proved remarkably difficult to assign a determinative role in vivo to any single antiviral immunological mechanism in recovery from a single viral disease, particularly since the immune response to the virus itself may frequently contribute to the pathology of the disease. Furthermore, if virus-induced immune responses are also directed against normal host components, this may set the stage for an autoimmune disease. In this context acute measles encephalomyelitis is of interest since in this disease autoimmune reactions against brain antigens have been observed and considered of pathogenetic importance. In this short review, virological and immunological findings of measles virus infections in a rat model in relation to autoimmune reactions will be presented and the mechanisms by which measles virus may alter host reactivity against self-antigens discussed. PMID- 8407254 TI - Yesterday, today and tomorrow in virology. PMID- 8407255 TI - T-cell autoimmunity in the central nervous system. AB - Studies of myelin basic protein (MBP)-specific T lymphocytes have been extraordinarily informative for several distinct aspects of (neuro) immunology. They indicated for the first time that myelin directed central nervous system (CNS) inflammation is mediated by specific autoaggressive T cells. This provided a conceptual basis for understanding the pathogenesis of anti-myelin autoimmunity in general. We assume at present that human multiple sclerosis (MS) is also caused by myelin-specific autoaggressive T cells, and that these T cells create the inflammatory changes typical for the generation of a mature demyelinating plaque. Large scale demyelination seems, however, not to be caused by purely cellular mechanisms. Second, studies using MBP-specific T-cell lines have critically contributed to elucidating the immune status of the CNS. They showed that the CNS is not completely secluded from the circulating immune cells. Activated, though not resting lymphocytes can pass through the endothelial blood brain barrier and migrate through the brain tissues. There, in the CNS, some glia components, microglia and astrocytes can be induced by T-cell cytokines to act as efficient antigen-presenting cells. Thus, the CNS is subject to immune surveillance, though to a specialized, adapted variant thereof. Finally, studies using MBP-specific T cells established for the first time the presence of potentially autoaggressive T cells within the normal immune repertoire. This implies counterregulatory regulation required to keep the autoreactive T cells in a safe state of rest. Most recently, the analysis of the intrathymic anti-MBP T cell repertoire may change our views on the generation of myelin-specific self tolerance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407256 TI - Cholecystectomy and oesophageal pathology: is there a link? AB - Symptoms persist in a significant proportion of patients following cholecystectomy, some of which may have an oesophageal aetiology. The oesophagus has not previously been studied in this patient group. In this study all patients who had undergone cholecystectomy over a four year period were invited for review and symptoms were documented. Oesophageal function was examined and compared with normal controls. Patients were subdivided into symptomatic and asymptomatic subgroups and their findings compared. Symptoms were present in 53 percent of the postcholecystectomy group. The mean (sem) DeMeester acid score was higher in the post-cholecystectomy group -20.6 (3.6) than in controls -6.7 (0.9) (p = 0.01). The incidence of oesophagitis and gastritis were also increased in this group. There was a trend towards increased reflux and oesophagitis in the symptomatic compared with the asymptomatic subgroup. Other findings confined to the post cholecystectomy group included nutcracker oesophagus in 4 and irritable bowel syndrome in 3. It is suggested that cholecystectomy may be associated with changes in oesophageal function which, in turn, may be associated with persistent symptoms. PMID- 8407257 TI - Renal replacement therapy in multiple myeloma and systemic amyloidosis. AB - Renal failure frequently complicates both multiple myeloma and systemic amyloidosis. Renal replacement therapy (RRT) may be poorly tolerated and its role in such patients is not clearly defined. Of fifty patients (26 males and 24 females) referred to a single centre because of renal failure associated with multiple myeloma or systemic amyloidosis 37 progressed to end-stage renal failure and 30 of these patients received RRT. Nine patients have been treated by CAPD, 13 by haemodialysis, and 8 patients have required both forms of dialysis. Overall one year and two year survival rates were 66% and 57% respectively. The median duration on RRT was 7.5 months (range 1-96 months) with a 51% one year, and a 46% two year survival rate. Of 7 patients with amyloidosis who underwent renal transplantation, 3 died within 6 months of transplantation. Undiagnosed cardiac involvement contributed to this early mortality. We conclude that renal replacement therapy is appropriate for some patients with multiple myeloma and systemic amyloidosis who develop endstage renal failure. Careful assessment and selection of patients is necessary prior to renal transplantation. PMID- 8407258 TI - Current alternatives to open cholecystectomy in the management of gallstones. PMID- 8407259 TI - Accuracy of computed tomography during arterial portography in the segmental localization of liver tumours. AB - The ability of computed tomography during arterial portography (CTAP) to identify and segmentally locate 20 hepatic tumours was prospectively assessed in 7 patients undergoing hepatic resection. All eight lesions greater than 2.5 cm in diameter were detected, but only two out of four and two out of eight of those measuring 1-2.4 cm and < 1 cm respectively. The overall segmental localization accuracy for detected lesions was 91%. Our results highlight the insensitivity of CTAP to hepatic lesions smaller than 1 cm in size, but have shown the accuracy of segmental localization for detected lesions. PMID- 8407260 TI - Atrial myxoma: national incidence, diagnosis and surgical management. AB - Despite being the most common benign intracardiac tumour with an excellent prognosis after surgical excision the incidence of atrial myxoma (except at autopsy) is unknown. We reviewed all patients admitted to the National Cardiac Surgery Unit (n = 26) with an atrial myxoma over a fifteen year period (1977 1991) to compile national incidence data and assess pre-operative diagnosis, management, surgical technique, and outcome. Preoperative symptoms were: congestive cardiac failure (12 patients), embolism (8 patients), constitutional (3 patients), asymptomatic (2 patients) and tachyarrhythmia (1 patient). The diagnosis was confirmed by 2D echocardiography alone in thirteen patients and by a combination of echocardiography and angiography in thirteen patients. At operation the site of the tumour was left atrial in 24 patients and bi-atrial in two patients. All cases were confirmed by histology. All patients made a good post-operative recovery, although one patient survived a pulmonary embolus and one patient developed a deep venous thrombosis. There has been one late death (five months after surgery) from a cerebrovascular accident. Serial echocardiography has revealed one recurrence to date (8 years after surgery). The surgical incidence of these tumours in the Republic of Ireland over the study period was 0.5 atrial myxomas/million population/year. Although rare atrial myxomas are the most important cardiac tumours to diagnose as the results from surgery are excellent. PMID- 8407261 TI - Clinical teaching: past--present--future. PMID- 8407262 TI - Prehospital resuscitation by Irish GP's: a preliminary report. AB - Two hundred and eleven Irish GP's responded to a questionnaire survey of their experiences of prehospital resuscitation. Seventy per cent had attempted CPR in a prehospital setting and a success rate of 21% was reported. A defibrillator had been used in the prehospital setting by 10% of the respondents with a reported success rate of 11%. Over 80% considered that they would benefit from further training in CPR and the use of a defibrillator. The implications of these results for the development of an Irish cardiac "chain of survival" are discussed. PMID- 8407263 TI - Six year follow up of forty five pregnant opiate addicts. AB - Forty five pregnant addicts had attended the National Drug Treatment Centre between 1984-1986. At that time they received intensive counselling, low dose Methadone maintenance and both ante natal and post natal care. Our aim was to follow these women six years later focusing on their drug use and outcome of their children. The women were followed up by chart review, individual interviews and liaison with the social and probation services. Results indicate that a high proportion of the women abused chaotically (50%). There is a worrying high incidence of HIV positive patients (53.4%) and a mortality figure of 15.5% (7). However only 13 women (28.6%) have had further children and 22 women (49%) are currently using some form of contraception. Only 23 women (51.1%) have had further contact with probation services. Five children (11.3%) are under formal care order and 4 children have become HIV positive in their own right. In conclusion, while these women have benefitted in certain areas e.g. family planning, contact with probation services, in other areas they have remained chaotic e.g. continued drug abuse or HIV risk taking behaviour. Thus the authors believe that future programmes should concentrate more directly on detoxification and rehabilitation after pregnancy. We also believe that because of the chaotic nature of these women some review of an "at risk" register for the children should be carried out. PMID- 8407264 TI - Eosinophilic fasciitis: a good response with conservative treatment. AB - A 52 year old man developed progressive painful swelling of both calves and difficulty walking. Physical examination showed asymmetrical localised swelling with induration and tenderness on palpation. Peripheral blood eosinophilia was noted. Biopsy of deep fascia and muscle showed typical features of eosinophilic fasciitis. He was treated with non-steroidal anti-inflammatory drugs and intensive physiotherapy. The clinical features had completely resolved six months later. PMID- 8407265 TI - A quantitative study of the perikaryon and the basal dendritic tree in rat hippocampus (CA1) pyramids following different experimental procedures. AB - A quantitative study of perikaryon and basal dendritic branching of hippocampal (CA1) neurons in male Wistar rats, in three experimental groups is reported. The groups of animals were exposed to treatment with either a tricyclic antidepressant, or a saline control injection or no handling. The hippocampal tissue was studied following Golgi-Kopsch impregnation and comparative measurements made using semi-automatic image analysis and light microscopy. Measurements of the extent of basal dendritic branching showed a significant difference (p < 0.01) between the saline-injected and unhandled control groups. The drug-injected and saline-injected groups had similar values although the drug injected group did not prove to be significantly different from the unhandled controls. No statistical difference was noted in the perikaryon size of the three experimental groups. These results are discussed with reference to the effects of sensory stimulation on neuronal plasticity. PMID- 8407266 TI - Multifactorial hypokalaemia in a beer drinker. PMID- 8407267 TI - Putrescine, polyamines, and N1-acetylpolyamine levels in retina, visual cortex and cerebellum of free-running mice kept under continuous light or darkness. AB - Putrescine, spermidine and spermine levels were detected in the retina, visual cortex, cerebellum and parietal cortex of CD1 mice exposed to 36h continuous light or darkness. Retinal putrescine and polyamine concentrations were found to be highest in dark-adapted mice, and the stimulation of dark-adapted retina with flicker illumination was also accompanied by a significant decrease in putrescine, spermidine and spermine levels. In visual cortex as well as in cerebellum spermidine and spermine contents were higher in dark-adapted mice in comparison to light-exposed animals, while in parietal cortex no significant change was found neither in spermidine nor spermine levels. In the brain areas studied flicker illumination produced no significant decreases in putrescine and polyamine contents. The total polyamines expressed as putrescine equivalents were noticeably decreased in retina, visual cortex and cerebellum of light-adapted mice. In the retina spermine/spermidine molar ratio was significantly higher than in dark-adapted mice. The administration of N1, N2-bis-(2,3-butadienyl)-1,4 butanediamine (MDL 72527) produced a strong decrease of retinal putrescine and spermidine concentrations in both dark-adapted and light-exposed mice, and in the retina of mice exposed to continuous light a significant decrease in the spermine level was also observed. According to the influence on polyamine reutilization, after the irreversible inhibition of polyamine oxidase by MDL 72527, in the retina N1-acetylspermidine and N1-acetylspermine accumulation was highest in light-adapted mice. On the contrary in visual cortex, cerebellum and parietal cortex the MDL 72527 administration produced a more marked decrease of putrescine and spermidine contents in mice kept in continuous darkness. PMID- 8407268 TI - A study on retinal photoisomerization catalyzed by a protein from the honeybee visual system. AB - The photoisomerization of all-trans retinal catalyzed by an isomerase purified from the honeybee retina is stereospecifically directed toward the formation of 11-cis retinal, the isomer required for rhodopsin regeneration. A model of the enzymatic reactions was developed, which fits the experimental data. In a particular experiment where irradiation was performed with a LASER light, the rate constants of the photoisomerization of all-trans retinal were dependent on the light intensity and were therefore greatly enhanced. PMID- 8407269 TI - Evaluation of a monoclonal antibody-based enzyme immunoassay for early detection of herpes simplex virus genital infection. AB - A monoclonal antibody-based enzyme immunoassay test for detection of herpes simplex virus (HSV) type-common antigen was evaluated in 40 women with vulvar lesion suspicious for genital HSV infection. The assay gave interpretable readings in 90% of the cases, with sensitivity and specificity of 100% and 92% respectively, a negative predictive value of 100%, and a positive predictive value of 75%. With a few limitations, the assay may be used for early diagnosis of genital HSV infection. PMID- 8407270 TI - Vaccination against hepatitis B in a general hospital in Israel: antibody level before vaccination and immunogenicity of vaccine. AB - We offered hepatitis B vaccine (Heptavax B) to 809 of the health care personnel of a 650-bed regional hospital; 290 accepted the offer. Anti-HBs measurement was done by enzyme immunoassay (AUSAB EIA, Abbott, UK) and expressed in mIU/ml. Seroconversion was determined at a level of 2.1 mIU/ml. Of 290 employees 58 (20%) were found positive for hepatitis B antibodies before vaccination. Of the laboratory technicians, 40.9% were found positive for antibodies before vaccination, as were 26.5% of nurses and 10.9% of physicians. Among vaccine recipients 35.8% responded after the first dose, 86.6% after the second and 92.7% after the third. Seventeen workers (7.3%) were nonresponders, of whom 14 received the whole vaccine series. There was no difference in immune responses to the vaccine between men and women. The present study confirms the relatively high prevalence of HBV infection in health care workers. Furthermore, vaccination of employees has been highly effective and well tolerated. The present data, therefore, support the introduction of active vaccination against HBV in health care workers in Israel. PMID- 8407271 TI - Intraperitoneal injection of platelet secretory products into mice increases macrophage uptake of oxidized low density lipoprotein. AB - Oxidized low density lipoprotein (LDL) (Ox-LDL) is taken up by macrophages at an enhanced rate and contributes to macrophage cholesterol accumulation and foam cell formation. Platelet secretory products have been shown to modulate the uptake of Ox-LDL by mouse peritoneal macrophages. This study is unique since mouse peritoneal macrophages were interacted with platelet conditioned medium (PCM, the supernatant that was obtained from collagen-treated washed human platelets) in the peritoneal cavity of the mice rather than in plastic dishes. Macrophages obtained from the peritoneal cavity of mice, 20 h after the injection of PCM (up to 30 micrograms of cholesterol/ml), demonstrated a substantial increment in the uptake of Ox-LDL. The effect of PCM demonstrated a dose- and time-dependent pattern. The cellular uptake of the lipoprotein, measured as the cellular Ox-LDL degradation and cholesterol esterification rates, was increased by up to 60% and 30% respectively in macrophages collected from PCM-injected mice in comparison to control mice. These effects were the result of PCM-induced increased affinity of Ox-LDL towards its receptor, and increased number of macrophage binding sites for Ox-LDL. Upon delipidation of PCM, only the protein fraction possessed the ability to increase the cellular uptake of Ox-LDL. Dialyzed PCM, which is deprived of low molecular weight substances, still expressed the stimulatory effect of PCM. Our results thus suggest that a protein like factor that is secreted from activated platelets can increase in vivo the ability of macrophages to take up Ox-LDL, as was also previously shown in in vitro studies. PMID- 8407272 TI - Heparin-induced thrombocytopenia and paradoxical venous thromboses. PMID- 8407273 TI - Does the gas mask jeopardize the fetus? PMID- 8407274 TI - Lyme disease acquired in Israel: report of a case and studies of serological cross reactivity in relapsing fever. PMID- 8407275 TI - Hepatitis B vaccination in hospital personnel: to B or not to B. PMID- 8407276 TI - Heparin-induced thrombocytopenia--pathogenesis and treatment. PMID- 8407277 TI - Unemployment and health. PMID- 8407278 TI - Ovarian cancer of epithelial origin--an update. PMID- 8407279 TI - Otto Loewy--discoverer of the mechanism of nerve conduction. PMID- 8407280 TI - Effect of aristolochic acid on arachidonic acid cascade and in vivo models of inflammation. AB - Recently, it has been reported that aristolochic acid inhibits phospholipase A2 (PLA2) in vitro and also decrease either oedema induced by snake venom and human synovial fluid PLA2. The aim of this research was to study the antiinflammatory activity of aristolochic acid and to investigate the effect of the alkaloid on the enzymes involved in the release of eicosanoids. Our results demonstrate that aristolochic acid is able to inhibit inflammation induced by immunological, immune complexes, and nonimmunological agents such as carrageenan or croton oil. We suggest that one mechanism of the antiinflammatory activity of alkaloid may be by directly blocking PLA2 catalyzed release of arachidonic acid. Moreover, this study demonstrates that aristolochic acid could also inhibit other steps involved in eicosanoids release such as cyclooxygenase and lipooxygenase pathways. PMID- 8407281 TI - Effect of mycophenolate mofetil (RS-61443) on cytokine production: inhibition of superantigen-induced cytokines. AB - The effects of the immunosuppressant mycophenolate mofetil (MPAM, RS-61443) on cytokine production at the single cell level were assessed using in vitro activated human mononuclear cells. Cytokine production was studied with UV microscopy of fixed and permeabilized cells stained with cytokine specific monoclonal antibodies (mAbs). The cytokines evaluated included interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10 interferon gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), TNF-beta, and granulocyte macrophage-colony stimulating factor (GM-CSF). MPAM exhibited a marked antiproliferative effect without cytotoxicity in all mononuclear cell cultures. Six to 24 hours after stimulation with the superantigen Staphylococcus aureus enterotoxin A (SEA), most cytokine production was unaffected by MPAM at therapeutic concentrations (10(-6) M), with the exception of GM-CSF. In contrast, by 48 h after antigen activation, MPAM significantly inhibited all studied cytokine production (p < 0.05). Cyclosporin A (CsA), used as a control at a concentration of 100 ng/ml, inhibited production of all studied cytokines, at all time points. Monokine production after lipopolysaccharide (LPS) stimulation was unaffected by MPAM. Similarly, the production of most of the cytokines studied after mitogen stimulation with phorbol ester (PMA) plus calcium ionophore (ionomycin) was not affected by MPAM, in comparison to CsA which demonstrated significant inhibition of all cytokines tested under these conditions. However, a late inhibitory effect on IL-3 production was seen by MPAM at 48 h after mitogenic stimulation. Further observations are required to explain the divergent results on cytokine production by MPAM in superantigen-activated and mitogen activated human mononuclear cells. PMID- 8407282 TI - Muscarinic cholinergic receptors on murine lymphocyte subpopulations. Selective interactions with second messenger response system upon pharmacological stimulation. AB - In this study the muscarinic cholinergic receptor capacity and affinity in murine lymphocyte sets and subsets and the biochemical responses obtained by the pharmacological stimulation of cholinergic receptors in these cells was determined by means of binding studies. Saturation assays with the specific radioligand for muscarinic cholinergic receptors ([3H]QNB) showed that B lymphocytes lack high affinity muscarinic cholinergic receptors, while the binding on T cells was a specific and saturable process. Lyt 2.2+ cells have a significantly higher number of receptors (Bmax) than L3T4+ cells; but the equilibrium dissociation constant (Kd) values obtained in both subpopulations did not differ significantly from one another, and resembled the Kd value of T lymphocyte populations. The specific receptor stimulation by carbachol caused a different intracellular signal transduction according the tested cell subtypes. The muscarinic cholinergic stimulation result in a significant inhibition of isoproterenol-stimulated adenylate cyclase system in T, L3T4+ and Lyt 2.2+ cells. On the contrary, Lyt 2.2+ cells were only able to respond to carbachol stimulus increasing cGMP levels and inositol phosphate formation while L3T4+ cells were unable to do it. Results show differences in the expression and in the ability of cholinergic receptors in sets and subsets of murine lymphocytes to trigger intracellular second-messenger systems. The differential receptor expression and the second-messenger response systems could be important to study the modulation of cellular immune response by cholinergic stimulation. PMID- 8407283 TI - Immunosuppression in adult female B6C3F1 mice by chronic exposure to ethanol in a liquid diet. AB - The overall objective of these studies was to characterize the effects of ethanol on the immunocompetence of adult female B6C3F1 mice. To obtain a significant suppression in the antibody response to SRBC, splenocytes from untreated mice had to be directly exposed to concentrations of ethanol from 0.3% to 3.0%, or to acetaldehyde at concentrations greater than 0.03%. We do not believe that these results are consistent with a role by a direct effect by either ethanol or its primary metabolite because these concentrations are higher than what could be obtained as reasonable blood levels. For in vivo exposure, we employed a pair feeding regimen which was based on a liquid diet containing 5% ethanol (v/v) that provided 36% of the caloric intake as ethanol. Our results indicated that there was a definite temporal relationship to the consequent suppression of the antibody response to SRBC in that no effect was observed after 14 days exposure, and that the magnitude of the suppression increased from 18% after 21 days to 70% after 42 days. We also monitored the liver for histopathology and observed that the ethanol-induced liver damage was restricted to steatosis (fatty liver), which was also manifested with time and which was most pronounced after 42 days exposure. In contrast to our results with the in vivo antibody response, we saw no effect on mitogen-induced proliferation by splenocytes from ethanol-treated mice. These results prompted us to measure in vitro antibody responses by splenocytes from ethanol-treated mice. We saw no suppression of the in vitro antibody responses to SRBC, DNP-Ficoll or LPS after any length of exposure to ethanol, and speculated that the basis for the suppression of the in vivo antibody response was an indirect consequence of exposure. We subsequently determined that when normal splenocytes were cultured in 5% serum from ethanol exposed mice (42-day group), there was a > 80% suppression relative to the serum from the pair-fed controls. As important controls for these studies, we have demonstrated that there was no difference between the responses of normal lymphocytes cultured in 5% normal mouse serum and in 5% serum taken from the pair fed restricted controls. A determination of the ethanol content in the serum from ethanol-exposed mice (42-day group) indicated that the amount of ethanol present in these cultures was < 0.003%.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8407284 TI - Comparison of effect of a panel of membrane channel blockers on the proliferative, cytotoxic and cytoadherence abilities of human peripheral blood lymphocytes. AB - The ability of human peripheral blood lymphocytes (PBMNC) to incorporate tritiated thymidine upon stimulation with concanavalin A, to perform natural cytotoxic (NK) activity and to form two types of spontaneous "rosettes" with sheep red blood cells (SRBC) upon in vitro treatment with membrane channel blockers of different specificity was estimated. All five channel blockers used, i.e., verapamil, nifedipine, diltiazem, saxitoxin and tetraethylammonium exerted a biphasic, dose-dependent effect on the Con A-induced proliferation of PBMNC. Verapamil, saxitoxin and nifedipine significantly increased the percentage of PBMNC forming spontaneously "active" rosettes with the SRBC. All channel modifiers tested for this ability decreased the percentage of PBMNC forming "late" rosettes. Natural killer activity was significantly decreased in the presence of micromolar concentrations of calcium antagonists as well as when the cells were treated with tetraethylammonium. The results of all four parameters tested were also compared with these obtained in a group of patients treated with 300 mg per day verapamil for two weeks preceding the tests. In this case we have found that treatment in vivo significantly decreased the response of PBMNC to mitogen stimulation, but it was without effect on either NK cytotoxicity or rosette formation. Cells of verapamil-treated patients did not show increased response to Con A in the presence of low concentration of the blocker in vitro. Also, they were not able to form more "active" rosettes when treated with 1-100 nM verapamil. We believe that apparent bi-directional pattern of action of calcium, sodium and potassium channel blockers on the proliferation and rosette formation of human PBMNC can be explained by two sets of events producing opposite results. First, we suggest that at respectively low concentrations of the blockers all of them generate uniform modulation of transmembrane electrical potential (depolarization) of lymphocytes. Depolarization promotes cell adhesion and is probably beneficial for early stages of the response to mitogen. At higher concentrations, this effect is overshadowed by other pharmacological actions of the blockers, of mostly inhibitory character. PMID- 8407285 TI - Increased production of atrial natriuretic peptide in the rat thymus after irradiation. AB - The rat thymus involutes when exposed to dexamethasone, and this phenomenon is accompanied by a striking increase of ANP localized in thymic macrophages. This study was designed to investigate whether a stimulus, that is X-rays, well known to also cause involution of the thymus, is followed by an elevation of ANP expression. Rats were irradiated with 4, 6 and 8 Gy, and after 4 days we found a dose-dependent increase (3-, 30- and 40-fold) of mRNA coding for ANP. The corresponding ANP precursor (ANP1-126) has been found elevated 4-, 10-, and 13 fold, respectively. Immunohistochemical analysis of irradiated (6 Gy) tissue revealed heavily IR-ANP positive cells in the cortex, which, due to their morphological and enzyme-histochemical characteristics, were identified as highly phagocytising macrophages. We suggest, thus, that regardless of the stimulus applied (DEX or X-rays) causing the vast cell death in the thymus, the increased ANP-expression is linked to the highly activated phagocytic system of the thymus under these conditions. PMID- 8407286 TI - Effect of immunological stimulation on the production of platelet-activating factor by rat peritoneal cells: its relevance to anaphylactic reactions. AB - The production of platelet-activating factor (PAF) by rat peritoneal cells was studied using as stimuli either monoclonal IgE, IgG1 or IgG2b anti-DNP (2,4 dinitrophenyl), and DNP-BSA. Peritoneal cells sensitized in vitro with any of these antibodies at concentrations higher than 10 nM and challenged with 1 microM DNP-BSA produced PAF. PAF production was also elicited by preformed IgE/ and IgG2b/DNP-BSA immune complexes, preferentially at a large antigen/antibody ratio. The production of PAF was unrelated to the activation of mast cells, since it occurred in populations depleted of mast cells by adherence to plastic dishes. Moreover, the release of [3H]serotonin from IgE-sensitized mast cells showed a time-course more rapid than PAF production and occurred in cells sensitized with IgE at concentrations lower than those required for PAF formation. In contrast, peritoneal cells sensitized with IgG1 and IgG2b failed to release [3H]serotonin. Rat peritoneal cells showed a significant ability to catabolize PAF by intracellular PAF-acetylhydrolase in view of both the amounts of enzyme activity assayed in cellular homogenates, and the 15-fold increase on controls of PAF quantities detected in peritoneal cells treated with phenylmethylsulfonyl fluoride (PMSF), a known inhibitor of PAF-acetylhydrolase. The PAF activity produced upon PMSF addition showed a retention time on reverse-phase HPLC which suggests structural identity to PAF produced by either immunological challenge or ionophore A23187. These data suggest that PAF formed during rat passive anaphylaxis reactions depends on the activation of mononuclear phagocytes. This production may be triggered by two types of low affinity receptors: Fc epsilon RII/CD23 and Fc gamma R. The ability of peritoneal cells to catabolize PAF by intracellular acetylhydrolase seems unaffected by immunological stimulation. PMID- 8407287 TI - Thymosin beta 4 synergizes with human granulocyte-macrophage colony-stimulating factor in maintaining bone marrow proliferation. AB - Recent evidence supports a role for thymosin beta 4 (T beta 4) in the inhibition of murine hematopoietic stem cell proliferation. This supposition results from studies in which the N-terminal tetrapeptide derived from native T beta 4 was administered to mice and appeared to prevent CFU-S recruitment into DNA synthesis. The importance of this observation was the concomitant ability of the tetrapeptide to prevent cytosine arabinoside (ara-C) toxicity in mice given LD50 doses of this drug. In the present study, we have extended these observations by demonstrating that whole synthetic T beta 4 is more effective than the N-terminal tetrapeptide in protecting mice from the toxicity of ara-C. This observation supports the hypothesis that T beta 4 is the biologically important parent molecule for this activity. To determine if inhibition of cell cycle progression also occurs in committed human bone marrow progenitors treated with T beta 4, we have investigated the effects of synthetic T beta 4 on proliferating and unstimulated enriched human bone marrow. In short-term liquid cultures studied sequentially over 1-7 days, T beta 4 failed to inhibit cell proliferation, but maintained the proliferative effect of granulocyte-macrophage colony stimulating factor (GM-CSF) on days following maximum stimulation (days 5-7). No effect was noted before the fifth day in culture, nor did T beta 4 exert any demonstrable effect in the absence of added GM-CSF. Any observable effect of T beta 4 required that it be present in the cultures on or before day 3 of GM-CSF stimulation. These results suggest that an additional effect of T beta 4 is the stimulation of a subpopulation of committed human bone marrow precursor cells to become more sensitive to the growth-promoting activity of GM-CSF, thereby enhancing myelopoiesis. It is of interest that the N-terminal peptide of T beta 4 is a shared sequence with tumor necrosis factor alpha, which is also known to have a similar stimulatory capacity. We, therefore, postulate that the growth enhancement noted in short-term cultures is mediated by the region containing these shared sequences. PMID- 8407288 TI - Parental alcohol and other drug (AOD) use and health beliefs about parent involvement in AOD prevention. AB - Because the family plays an important role in predicting alcohol and other drug (AOD) use among youth, prevention efforts must begin in early childhood and involve parents. The purpose of this study was to determine differences between parent AOD users and nonusers in relation to health beliefs about parent involvement in AOD prevention with preschool children. A convenience sample of 200 Head Start parents in East Central Indiana completed self-report instruments measuring Health Belief Model constructs. Almost half (45%) reported alcohol use, over half (54%) smoked cigarettes, and 11% admitted using illicit drugs. Compared to nonusers, illicit drug users were more likely to perceive their children as susceptible to future AOD use, but less likely to view AOD use by their children as serious. Compared with nonsmokers, tobacco users were more likely to perceive their children as susceptible to future AOD use and had less interest in maintaining health through early prevention activities. PMID- 8407289 TI - Childhood sexual abuse: sources of trauma. AB - Many American women who were sexually abused as children seek mental health services to help them heal from their abuse. An appreciation of the varied sources of trauma that may stem from a sexual abuse experience may guide clinicians in facilitating a meaningful discussion with survivors of the ways in which their childhood development and their current lives have been influenced by their sexual abuse. Therefore, the goal of this study was to provide a beginning delineation of possible sources of trauma in the abuse situation, based on the retrospective reflections of women who have survived abuse. One hundred and eighty-six survivors were asked to identify the most traumatic aspects of their abuse experience. A content analysis was performed on their written responses, and the following eight categories, reflecting different sources of trauma, were identified: abandonment, powerlessness, violence, betrayal, guilt and shame, loss of self, loss of childhood, and impact on sexual adjustment. Possible treatment implications and suggestions for future research are discussed. PMID- 8407290 TI - Relationship between psychiatric diagnosis and patient aggression. AB - Patient aggression is a common problem on psychiatric units. Recent research has focused on the relationship between aggressive behavior and various patient characteristics, including psychopathology. The purpose of this study was to examine the relationship between diagnostic groups and particular types of patient aggression in a state psychiatric hospital. A secondary analysis was performed on existing data. There was no significant difference noted among DSM III diagnoses in total patient aggression. However, there was a significant difference between individuals with organic disorders and individuals with paranoid schizophrenia in the category of physical aggression against other people. Improved ability to identify a particular diagnostic group with the potential for aggression will strengthen efforts to decrease the risk of patient aggression in psychiatric facilities. PMID- 8407291 TI - Chinese nurse stress in Taiwan, Republic of China. AB - An exploratory study was conducted to understand the Chinese nurses' perceptions of their work stress in Taiwan, Republic of China (ROC). Data were based on the written answers to two open-ended questions from the randomly selected Chinese nurses working at 3 of the 10 first-ranked teaching hospitals that had 900-2,000 hospital beds. The findings showed that stressors in work situations for Chinese nurses were similar to those of their Western colleagues in four categories: nursing care related to patient condition, interpersonal relationships, workload, and opportunity for promotion. In addition, the Chinese respondents especially identified pressure in the role of a unit educator as stressful. Any troubled interpersonal relationship of the Chinese nurse may have been experienced as a greater source of pressure than nurses in other cultures. It is suggested that there might be some similar stressors in the nurse work situation when comparing the Chinese nurse working in the modern hospital in Taiwan to the nurses in large hospitals in the Western culture. Yet the difference between these nurses was the greater emphasis the Chinese nurses placed on the value of advanced study and interpersonal harmony. PMID- 8407292 TI - Dis-ease between nursing and feminism: nurses caring for one another within a feminist framework. AB - Mental health professionals have the experience and skills necessary to provide care and support to colleagues and peers in these troubled and stressful times. Yet, traditionally nurses have not been supportive of one another. Not receiving much interest from the feminist movement and not willingly to embrace feminist social activism, the nursing profession has suffered as it continues to embrace racist, classist, sexist, and homophobic outdated theories and research practices. Without a collective voice which unites all nurses in a common struggle, issues related to mental health nursing will continue to be viewed in a fragmented way. Until feminist research, theory, and practice become central to the profession's ideology, nurses will continue to be divided among themselves, unable to provide the necessary caring for one another. PMID- 8407293 TI - Current morale issues that impede the caregiving process of substance abuse/addictions nurses. AB - A telephone survey was conducted with 25 nurse managers of substance abuse/addictions programs in a metropolitan area to ascertain what morale issues impede the caregiving processes of their nursing staff. Five issues were repeatedly identified by all 25 nurse managers. Those issues were: (a) lack of credibility and respect for the nurse in the eyes of other members of the multidisciplinary team, (b) lack of understanding of the role of nurses in the substance abuse/addictions program on an inpatient unit; (c) lack of adequate preparation for the role, (d) lack of acceptance of the nursing role as defined by nursing, and (e) the care of patients with concurrent diagnoses. This article analyzes these issues from the perspective of reasons for existence and possible solutions. PMID- 8407294 TI - Duty, truth, and whole human beings. PMID- 8407295 TI - Culture, healing, and professional obligations. PMID- 8407296 TI - The impending collapse of the whole-brain definition of death. PMID- 8407297 TI - Terra es animata on having a life. PMID- 8407298 TI - Pursuing a peaceful death. PMID- 8407299 TI - The fears of the dying. PMID- 8407300 TI - The fears of the dying. PMID- 8407301 TI - Cannon to right of them. PMID- 8407302 TI - Cannon to right of them. PMID- 8407303 TI - Cannon to right of them. PMID- 8407304 TI - Cannon to right of them. PMID- 8407305 TI - Gridlock on the Oregon trail. PMID- 8407306 TI - Medicine and human rights. A proposal for international action. PMID- 8407307 TI - Clinical utility of intraoperative frozen section diagnosis in head and neck surgery: a quality assurance perspective. AB - The technical and professional issues that affect the clinical utility of intraoperative frozen section diagnosis in head and neck surgery are addressed from a quality assurance perspective. The most common reasons for sampling errors and interpretive errors are discussed. We offer several recommendations for head and neck surgeons and pathologists which will optimize the clinical utility of intraoperative frozen section consultation. PMID- 8407308 TI - Therapeutic lymphadenectomy in melanomas of the head and neck. AB - This retrospective study describes patients with loco-regionally metastasized melanoma of the head and neck. All patients underwent a therapeutic lymph node dissection. The 3-year survival rate was 35%. Duration of disease-free interval, number of lymph nodes involved, and extent of neck dissection proved of no influence on survival rates. Locoregional recurrence occurred in 10 patients and always proved to be a sign of systemic dissemination. Ultimately, 19 patients developed systemic disease. More than 40% developed cerebral metastases and the cerebrum was the second most involved site. As cerebral involvement occurs often in head and neck melanoma, a computed tomography or magnetic resonance imaging scan of the brain is recommended in the routine work-up before lymph node dissection. Furthermore, because the extent of the surgical procedure had no influence on local recurrence rate and overall survival, a selective approach, preserving functional structures, is advocated. PMID- 8407309 TI - Multivariate analysis results of radiotherapy for laryngeal cancer. AB - We assess the impact of radiotherapy in the treatment of laryngeal cancer and evaluate the value of the standard dose (linear quadratic plus time model) and other variables to predict tumor control and survival. Between 1972 and 1989, 80 patients with laryngeal cancer received comprehensive radiotherapy. Patients with stage I laryngeal glottic cancer (T1-N0-M0) were excluded from this study. Mean follow-up was 15 months (range 4 to 181). The mean age was 64.8 years (range 40 to 92). Standard dose varied from 32.65 to 81.81 Gy (mean 66.78). The 5-year overall survival and tumor-specific survival rates were 44.9 +/- 5.8% and 51.4 +/ 5.9%, respectively. Five-year local control and locoregional control rates were 66.4 +/- 5.7% and 61.9 +/- 5.8%, respectively. Multivariate analysis showed that local control was significantly predicted by T stage (p = .032), but not by standard dose (p = .906). Independently significant factors predicting tumor specific survival included stage (p = .006), site (p = .019), and age (p = .001). Local control and survival were significantly predicted by the TNM-staging classification. The standard dose did not predict local recurrence or survival. PMID- 8407310 TI - Secondary microvascular tongue reconstruction: functional results. AB - Between 1978 and 1991, 56 microvascular composite flaps were used for oromandibular reconstructions: 15 for primary total and subtotal tongue reconstruction and five for secondary major tongue reconstruction. The delayed reconstructions were performed to improve the oral and pharyngeal phases of swallowing. Using a floor of the mouth composite bone grafting technique to reposition the tongue and obliterate the oral dead space intraoral food transport was improved (three of five patients), but aspiration persisted (three of four patients). When compared with 10 patients evaluated for primary total and subtotal tongue reconstruction the primary reconstruction group showed superior swallowing (eight dynamic oral transport, no aspiration) and speech results. The poor results of secondary reconstruction are attributed to scarring and irreversible damage to remaining functional muscles involved in protecting the laryngeal aditus. PMID- 8407311 TI - Laryngeal cancer in Colorado. AB - Previous studies of laryngeal cancer have shown a decrease in the male-to-female ratio and that the sex ratio in glottic tumors is higher than that for other laryngeal sites. The purpose of this study was to characterize and identify changes in the demographics, laryngeal site predilection, geographic distribution, trends in tumor stage at diagnosis, and surgical caseload distribution in Colorado. The Colorado Central Cancer Registry for the years 1979 to 1990 was reviewed for cases of laryngeal cancer. The data were analyzed using chi-square and gradient-in-proportions methods. One-thousand two-hundred sixty five (1265) cases were identified with a male-to-female ratio of 4.3:1. There was a significant trend of an increasing proportion of cases attributable to women. Glottic carcinoma accounted for 50.1% of cases, with a male-to-female ratio of 7.6:1. The Denver metropolitan area accounted for 57% of all cases, which was not disproportionate to the population. From 1981 to 1990 there was a significant decrease in the proportion of stage I cases and an increase in stage II cases. The teaching hospitals associated with the University of Colorado were responsible for performing 44.5% of all surgery for laryngeal cancer. The results indicate that laryngeal cancer in Colorado shares similar epidemiologic characteristics to those reported in other studies. Of most concern is the increasing proportion of women with laryngeal cancer and the decrease in cases diagnosed at stage I. PMID- 8407312 TI - Squamous cell carcinoma of the hypopharynx--analysis of treatment results. AB - The results of surgical treatment and causes of failure in 109 patients with squamous cell carcinoma of the hypopharynx were analyzed. The 5-year survival was significantly related to the stage of the disease (stage I, 74%; stage II, 63%; stage III, 32%; and stage IV, 14%) and the preoperative nodal staging (N0, 57%; N1, 28%; N2, 6%; and N3, 0%), but not related to the extent of tumor resection. The local control rate was 86% and the majority of the local recurrences occurred at the upper resection margin. There was no significant difference in the local recurrence rate between those patients who had pharyngolaryngoesophagectomy and those who had pharyngolaryngectomy. However, the resection-associated complications (bleeding, cardiac arrhythmia, pulmonary complications) were more frequent when total esophagectomy was performed. Reconstruction-associated complications such as wound infection and anastomotic leakage occurred less often after gastric pull-up reconstruction as compared with the use of myocutaneous flap, but was associated with more serious outcome after pharyngo-gastric anastomosis. Thirty-seven patients (34%) had neck node recurrences and these were significantly related to the preoperative nodal staging (N0, 20%; N1, 37%; N2, 48%; and N3, 83%). Additional primary cancers occurred in 17% of the patients and was a significant cause of death in those patients who survived more than 2 years. PMID- 8407313 TI - Primary site management following induction chemotherapy with cis-platinum, 5 fluorouracil, and leucovorin. AB - A phase II clinical trial was initiated in 1987 to evaluate a new induction regimen of cis-platinum, 5-fluorouracil, and leucovorin (PFL) for patients with stages III-IV squamous cell carcinoma of the head and neck. Ninety patients were treated and followed for a median duration of 18 months. The median age was 55 and 87% of the patients had stage IV disease. The rates of complete and overall clinical response following three cycles of PFL were 57% and 80%, respectively; the rate of complete response at the primary site was 72%. Eighty-four percent of patients were treated to the primary site with radiation alone (median dose 68 Gy in daily 1.8-Gy fractions) irrespective of the location of the primary site or initial T-stage. The acute tolerance to full-course radiation following PFL was acceptable. The actuarial rate of primary site control for patients treated with radiation was 67% at 36 months. An important prognostic indicator for primary site control was a complete clinical response to induction PFL. For patients who achieved a complete response, radiation or surgery followed by radiation controlled primary site disease equally well at 70%. Patients with a partial response did less well. For these patients, surgery and radiation appeared slightly better than radiation alone. PMID- 8407314 TI - Effects of maneuvers on swallowing function in a dysphagic oral cancer patient. AB - This study examined the effects of three swallow maneuvers: (1) the supraglottic swallow (voluntary closure of the vocal folds prior to the swallow); (2) the super-supraglottic swallow (airway closure at the level of the arytenoid to base of epiglottis); and (3) the Mendelsohn maneuver (voluntary prolongation of laryngeal elevation and cricopharyngeal opening during swallow) on swallow functioning in a 47-year-old patient who underwent right composite resection for a squamous cell carcinoma of the right retromolar trigone. All maneuvers were employed during the same videofluorographic (VFG) swallow study conducted 6 months after the patient's surgery. Biomechanical analysis of 3-mL swallows defined the extent and timing of tongue base retraction to the posterior pharyngeal wall, laryngeal elevation, laryngeal closure and cricopharyngeal opening during swallows with and without maneuvers. Airway closure duration was prolonged during supraglottic and super-supraglottic swallows, but aspiration was not eliminated. Use of the Mendelsohn maneuver improved coordination and timing of pharyngeal swallow events, including timing of posterior movement of the tongue base to the pharyngeal wall in relation to airway closure and cricopharyngeal opening, with elimination of aspiration. The Mendelsohn maneuver compensated for anatomic and physiologic changes in the oropharyngeal swallow and enabled reinstatement of safe oral intake in this surgically treated head and neck cancer patient who was previously unable to take nutrition orally. PMID- 8407315 TI - Sedimentation rate and serum thymidine kinase activity: prognostic factors in squamous cell head and neck cancer. AB - Identification of prognostic factors in squamous cell head and neck cancers involves analysis of highly diverse clinical and biological parameters. This study analyzed the prognostic value of clinical variables (age, sex, tumor site, stage) and biologic parameters (squamous cell carcinoma antigen [SCC], serum thymidine kinase activity [TK], fibrin, sedimentation rate [SR]) at the time of diagnosis of squamous cell carcinoma of the head and neck (oral cavity, oropharynx, hypopharynx) in 189 patients. Among the clinical variables investigated, UICC stage III-IV disease (p < .0002), a hypopharyngeal site (p < .02), and age over 60 years (p < .01) were all associated with a poor prognosis. Similarly, analysis of biological blood variables allowed definition of cut-off values above which the prognosis was poor: SCC 2.5 ng/mL (p < .01), fibrin 3.5 g/L (p < .01), TK 7 IU/L (p < .0005), and SR 15 mm per first hour (p < .0000). Cox regression analysis of overall survival identified the UICC stage (p < .000), the SR (p < .001), and serum TK (p < .02) as the main independent prognostic factors. A separate study on a small number of head and neck cancer patients revealed higher TK levels in malignant squamous cell carcinoma tissue than in adjacent healthy tissue. PMID- 8407316 TI - Platysma myocutaneous flap for head and neck reconstruction in cats. AB - Squamous cell carcinoma is the fourth most common neoplasm in cats. As in man, local resective surgery of stage III and IV carcinoma often results in recurrence related to compromised margins. Radical resective procedures may be performed when reconstructive techniques are available to restore cosmesis and function. A platysma myocutaneous flap that was based on a cutaneous branch of the caudal auricular artery and vein was developed to fulfill this requirement. Control flaps, which included ligation and division of the caudal auricular artery and vein, were similarly developed on the contralateral aspect of the neck. Mean survival of all platysma myocutaneous flaps (86.7%), compared with control flaps (62.9%), was significantly different (p < .05). Flaps grouped in lengths of 6, 9, and 12 cm had mean survival lengths of 93.8%, 81.9%, and 84.4%, respectively. The mean survival length of flaps measuring 12 cm in length was significantly different (p < .05) compared with flaps measuring 6 and 9 cm. On the basis of the results of this study, the platysma myocutaneous flap based on a cutaneous branch of the caudal auricular artery and vein may be a source of tissue for reconstructive procedures of the head and neck in cats. PMID- 8407317 TI - Patterns of spread of squamous cell carcinoma to the ramus of the mandible. AB - Patterns of spread of squamous cell carcinoma to the ramus of the mandible have been studied. In nonirradiated mandibles, tumor in the ramus was found to have spread in continuity from tumor within the body. In 22 of 27 specimens (81.5%) with established invasion of the molar region of the body, tumor spread was either confirmed to the body or to the body and the anterior part of the ramus. The remaining five cases showed extensive spread within the ramus. It was concluded that a more conservative approach to resection of the ramus may be safe on pathologic grounds in the nonirradiated mandible. In postirradiation mandibles, direct invasion through cortical bone was seen in addition to direct spread from the body. Tumor spread to the ramus was seen in nine of 12 specimens (75%), and in each case was both diffuse and extensive. It was concluded that a conservative approach to resection of the ramus is not safe on pathologic grounds after radiotherapy. PMID- 8407318 TI - Surgical management of chronic parotitis. AB - A series of 17 consecutive parotidectomies for chronic sialoadenitis is presented. This comprises 10% of all parotidectomies performed by one surgeon over the 5-year period between 1987 and 1991. In 16 patients, symptoms were relieved by surgery. The extent of surgery was guided by the clinical findings. In this series, near total and superficial parotidectomy were equally efficacious and no patient suffered permanent facial nerve dysfunction. Surgery is a safe and effective treatment for parotitis. PMID- 8407319 TI - Vocal cord paralysis from prostatic carcinoma metastasizing to the larynx. AB - According to 1992 Cancer Statistics, prostate carcinoma is once again predicted to be the most common cancer in men, exceeding the incidence of lung cancer. In American men, this cancer is estimated to be the second most frequent cause of cancer deaths by site. Although metastases have been reported in practically every organ in the body, prostatic cancer most often metastasizes to bones, regional lymph nodes, and viscera. Although secondary involvement of the larynx by malignant neoplasms arising in contiguous structures is well known, metastatic cancer to the larynx from a prostate carcinoma is rare. This report discusses a unique case that presented with hoarseness resulting from vocal cord paralysis. The diagnosis of the tumor was confirmed by immunoperoxidase stains for prostate specific antigen. PMID- 8407320 TI - Mesenchymal chondrosarcoma of the jaw--report of a case and review of 41 cases in the literature. AB - A case of mesenchymal chondrosarcoma in the mandible of a 35-year-old woman is presented. In a thorough survey of the world literature, 41 cases of mesenchymal chondrosarcoma occurring the jaw are reviewed. PMID- 8407321 TI - Carcinoma of the tonsil. AB - Although the experts acknowledge that there is no conclusive evidence linking secondhand smoke to head and neck cancer, a recent report by the Environmental Protection Agency classifies secondhand smoke as a group A carcinogen. There is strong evidence linking it to carcinoma of the lung. Whereas you may not be able to tell your patient that the same cause and effect is present for head and neck cancer, it is the editor's belief that this will one day be proven. Three experts agreed to treat this patient with surgery followed by full-course radiotherapy, although the surgical approaches differed. They included a marginal mandibulectomy, radical neck dissection, and plating of the remaining mandible (Dr. Strome) and a composite resection (Drs. Ward and Johnson). For reconstruction, options included a modified FAMM flap or a split-thickness skin graft (Dr. Strome), tongue flap or pectoralis major myocutaneous flap (Dr. Ward), or a split-thickness skin graft (Dr. Johnson). One consultant suggested resecting the neck mass and treating the primary tumor and neck with radiotherapy. A dental consultation is in order prior to radiotherapy (Dr. Goepfert). With regard to this woman's mental status, all the experts called for counseling. The husband should be included in the discussions (Dr. Strome and Ward) and consideration should be given to the Women's Right Advocacy Group (Dr. Johnson). PMID- 8407322 TI - [Migraine and allergy]. PMID- 8407323 TI - [Acquired benign myofibroma of the skin (adult myofibroma)]. AB - Adult myofibroma is an acquired skin tumour that is histologically identical to infantile myofibroma. Histologically, four tumour variants can be distinguished: leiomyoma type, cellular spindle cell type, haemangiopericytoma or glomus type, and biphasic type. Practical help in diagnosis is supplied by the hamartomatous composition of the leiomyoma-type, hyalinized spindle cell whorls, endothelium lined haemangiopericytoma-like vascular channels, and desmin-negative, muscle actin-positive myofibroblasts, which may show spindle cell or undifferentiated mesenchymal cell differentiation. The tumours clinically present as firm, occasionally bluish cutaneous or subcutaneous nodules. Adult myofibroma should be recognized as a distinct benign neoplasm, most probably of vascular origin. Clinicians and dermatopathologists must be aware that lesions identical to those of infantile myofibroma can occur in adults. PMID- 8407324 TI - [Follow-up of atopic dermatitis after early childhood]. AB - A total of 106 patients, 57 women and 49 men, had had atopic dermatitis even in infancy and were monitored by means of questionnaires at a mean age of 23.5 years. Items of interest were the course of the dermatosis, influences on its course and the frequency of respiratory allergic manifestations. The persistence rate of atopic dermatitis was 60.4%. It was possible to identify ten different courses of atopic dermatitis. Only 11.3% of these patients had suffered from the eczema exclusively during infancy. In 27.4% of cases the patients were free of manifestations for an average of 9 years, after which they had a new attack of the illness. Almost one third of the patients showed a continuous type of development of their atopic dermatitis, which had persisted since infancy. The most common localizations were the antecubal fossa (48.4%), the fingers (45.3%) and the face (37.5%). The course of atopic dermatitis was influenced by the seasons in 71% of these patients, and 59.4% observed a connection between psychic stress and the exacerbation of skin conditions or a new attack of the dermatosis. As adults, 59.4% of all patients suffered from one or more respiratory allergic manifestation: 41.5% had pollinosis, 24.5% allergic rhinitis, and 25.5% bronchial asthma. Respiratory allergies disappeared in 24 cases (19.8%), bronchial asthma at a mean age of 13 years, and pollinosis at 15.3 years and perennial rhinitis at 16 years on average. PMID- 8407325 TI - [Stigmas, symptoms and diseases of the skin in musicians]. AB - There are numerous characteristic skin symptoms that are common in musicians and are usually harmless marks of their profession. There are, however, many less frequent and more serious skin diseases that are due either to mechanical irritation and other physical noxae or to allergic and non-allergic contact reactions. Professional musicians can be affected by other skin diseases that cause serious problems and ultimately put an end to their career. This applies especially to lip diseases in woodwind and brass players. PMID- 8407326 TI - [Cutaneous microcirculation in pretibial necrobiosis lipoidica. Comparative laser Doppler flowmetry and oxygen partial pressure determinations in patients and healthy probands]. AB - Necrobiosis lipoidica (NL) is a chronic, skin disease usually localized pretibially and often associated with diabetes mellitus. Primary vascular disturbances are considered pathogenetic for NL. In order to determine microcirculatory alterations in "idiopathic" NL linked with neither arterial hypertension nor diabetes mellitus, we performed simultaneous measurements of laser-Doppler flux (LDF) and oxygen tension (pcuO2) in 10 non-diabetic patients with NL and in 10 age- and sex-matched healthy volunteers. We examined the centre of the pretibial NL plaque, its border and the non-affected skin of the proximal lower leg at probe temperatures of 36 degrees C (flux) and 37 degrees C (pcuO2). Corresponding sites and temperatures were chosen for the controls. In addition, the degrees of hyperaemia caused by arterial occlusion (3 min) and during local heating (42 degrees C) were continuously measured. The initial pcuO2 values were found to be lowered, in contrast to increased fluxes within and around the NL lesions. In addition, responses of both flux and pcuO2 to the hyperaemic stimuli were weaker than in the corresponding skin of the healthy controls. Only minor differences from controls or none at all were found in clinically unaffected lower leg skin of the patients. Our results indicate a local alteration of microcirculation in patients with "idiopathic" NL that occurs independently of diabetes. PMID- 8407327 TI - [CHILD syndrome. Case report of a rare genetic dermatosis]. AB - We report on a 2-month-old girl who developed unilateral ichthyosiform erythrodermia on the right side of her trunk and the right leg 1 week after birth. In addition, there was hexadactyly of her right hand. Apart from that, close physical examinations did not reveal any other defects. We diagnosed this condition as CHILD syndrome (congenital hemidysplasia with ichthyosiform nevus and limb defects). PMID- 8407328 TI - [Pemphigus herpetiformis]. AB - Pemphigus herpetiformis is an unusual variant of pemphigus. Clinically it can resemble Duhring's disease, while histopathological examination and, in particular immunofluorescence yield findings diagnostic for pemphigus. As pemphigus herpetiformis responds quite well to sulfones, recognition of this rare disorder is of practical relevance. In the present report, a case of pemphigus herpetiformis with an unusually young age of onset is presented. In addition, clinical features, results of histopathological examination and immunofluorescence, differential diagnosis and treatment are discussed. PMID- 8407329 TI - [Pseudoacanthosis nigricans simulating papillomatosis confluens et reticularis (Gougerot and Carteaud)]. PMID- 8407330 TI - [Amalgam and effluvium in women]. PMID- 8407331 TI - [External treatment of atopic eczema with gamma-linolenic acid?]. PMID- 8407332 TI - [2nd Workshop of the Study group of the ADF: Flow cytometry in dermatology. 16 September 1992 in Bonn]. PMID- 8407333 TI - [Meeting report. Report on the 6th meeting of the Epidemiology, Biostatistics and Informatics Study Group with the Dermatohistology Study Group, 22 and 23 January 1993 at the Tubingen University Dermatology Clinic]. PMID- 8407334 TI - [Bee and wasp venom allergy]. PMID- 8407335 TI - "Does inappropriate use explain small-area variations in the use of health care services?" A critique. PMID- 8407336 TI - Calculating the probability of rare events: why settle for an approximation? AB - OBJECTIVE: Health services researchers often need to compute the probability of observing a certain number of events when only a few such events are expected. Our objective is to show that the standard approaches (Poisson, binomial, and normal approximations) are inappropriate in such instances, and to suggest an alternative. DATA SOURCES: Patients undergoing cholecystectomy (34,234) in 465 California hospitals in 1983 are used to demonstrate the biases arising from various methods of calculating the probability of observing a given number of deaths in each hospital. Similar data from other procedures and diagnoses with lower and higher mortality rates are also used for illustration. STUDY DESIGN: The computational methods to derive probabilities using the Poisson, normal, simulation, and exact probabilities are discussed. Using a previously developed risk factor model, the probability of observing the actual number of deaths (or more) is calculated given the expectation of death for each patient in each hospital. Results for the four methods are compared, showing the types of random and systematic errors in the Poisson, normal, and simulation approaches. DATA COLLECTION: Routinely collected hospital discharge abstract data were provided by the California Office of Statewide Planning and Development. PRINCIPAL FINDINGS: The Poisson and normal approximations are often biased substantially in calculating upper-tail p-values, especially when the expected number of adverse outcomes is less than five. Simulations allow unbiased calculations, and the degree of random error can be made arbitrarily small given enough trials. Exact calculations using a simple recursive algorithm can be done very efficiently on either a mainframe or personal computer. For example, the whole set of cholecystectomy patients can be assessed in less than 90 seconds on a Macintosh. CONCLUSIONS: Calculating the probability of observing a small number of events using standard approaches may result in substantial errors. The availability of a simple and inexpensive method of calculating these probabilities exactly can avoid these errors. PMID- 8407337 TI - Could distance be a proxy for severity-of-illness? A comparison of hospital costs in distant and local patients. AB - OBJECTIVE: We test the hypothesis that hospital costs, after adjusting for DRG mix, are higher in distant patients than in local patients. DATA SOURCES AND STUDY SETTING: Data were obtained from the Washington State Commission Hospital Abstract Reporting System (CHARS) and included all patients discharged from 15 metropolitan hospitals in the state of Washington during fiscal year 1987 (N = 181,072). STUDY DESIGN: Distant patients were initially defined as those patients residing outside a 15-mile radius of the hospital from which they were discharged; all other patients were considered local. Distance was determined using the patient's residence zip code. Hospital charge, calculated for all patients regardless of payer, served as a proxy for cost and was adjusted using the DRG weight. PRINCIPAL FINDINGS: Average charge (adjusted for DRG weight) was higher for distant patients in all but two hospitals. Overall adjusted charge for distant patients was 15 percent higher (p < .001). This finding persisted when different distances were used to dichotomize distant and local patients. When the 20 most common DRGs were examined individually, little charge difference was found in surgical DRGs that require tertiary center services (tertiary DRGs) and in those DRGs with both moderate and predictable resource use (routine DRGs); the charge difference seemed most prominent in those DRGs with a wide array of possible resource use (heterogeneous DRGs). CONCLUSIONS: Results suggest that patients traveling long distances use more resources and incur higher hospital charges than local patients. This is not accounted for in prospective payment. We postulate that distance might serve in part as a proxy for severity-of-illness. PMID- 8407338 TI - Interorganizational exchanges as performance markers in a community cancer network. AB - OBJECTIVE: This study examines how "strategic partnerships" between community based consortia of oncologists and hospitals (CCOPs) and clinical cooperative groups emerge, develop, and influence patient accruals (i.e., the number of patients enrolled in clinical trials) over time. DATA SOURCES AND STUDY SETTING: Study analyses are based on 65 pairwise relationships that 38 CCOPs established with eight clinical cooperative groups in September 1983 and maintained through February 1989. Data are drawn from grantee applications and progress reports. STUDY DESIGN: The study examines how different types of CCOP-cooperative group exchange relate to one another and to CCOP patient accruals over six time points. Key independent variables include resource dependence, information exchange (i.e., meeting attendance and committee membership), and protocol exchange (i.e., the number of different protocols used). DATA COLLECTION METHODS: Data extracted from secondary sources were entered in a data base. PRINCIPAL FINDINGS: The number of CCOP physicians and support staff who attend cooperative group meetings during the first two years of a clinical research partnership has a significant influence on meeting attendance and protocol use in later years. Two-thirds or more of the variance in patient accruals at each time point can be explained by the number of different protocols used and the number of CCOP representatives serving on cooperative group committees (or attending cooperative group meetings). CONCLUSIONS: The findings highlight the importance of historical relationships and anticipated resource dependence in shaping initial exchange patterns. They also suggest that strategic partnerships need to emphasize structures and processes that encourage early involvement in collaborative activities and that reward participants for maintaining high levels of interaction. PMID- 8407339 TI - Effects of the relative fee structure on the use of surgical operations. AB - OBJECTIVE: The goal is to develop a theoretical and empirical framework for investigating how the demand for an operation may be affected by the fee for the operation (the own-price) and by fees for other services provided by surgeons in the same specialty (the cross-price). The theory suggests an empirical test of whether surgeons create demand for surgery. DATA SOURCES AND STUDY SETTING: The study examines the use of 11 frequently performed surgical operations by elderly Medicare enrollees in a cross-section of 316 U.S. metropolitan areas. Medicare physician claims and enrollment files for 1986 are the principal sources of data. STUDY DESIGN: Using econometric methods, a structural demand equation modified to include the own-price and the cross-price is estimated for each study operation. PRINCIPAL FINDINGS: The theory suggests that the utilization response to changes in fees may differ among operations depending on whether demand creation occurs and on the interplay of distinct own-price and cross-price effects. However, the results of the empirical analyses are inconclusive regarding the most appropriate economic model of surgical utilization. Both neoclassical behavior and demand creation are observed, but technical limitations of the analyses, including the cross-sectional design of the study, preclude definitive inferences. CONCLUSIONS: Despite the lack of definitive empirical results, the study has several implications for future research regarding the effect of changes in fees on surgical utilization. In particular, future studies should consider the roles of distinct own-price and cross-price effects, examine the importance of the supply demand balance in physician services markets, and assess whether typologies of operations that are based on the strictness of their clinical indications predict the appropriate economic model of utilization. PMID- 8407340 TI - Multiple-site physician practices and their effect on service distribution. AB - OBJECTIVE: This study explores the impact of multiple-site practices on the distribution of physician services within a medical service region. DATA SOURCES AND STUDY SETTING: A questionnaire was mailed to all urologists (100 percent response rate) practicing in north central Connecticut (the Hartford medical service area) and adjacent communities in September 1990. Data on community characteristics were obtained from the 1990 U.S. census and state government documents. STUDY DESIGN: Descriptive statistics and maps were used to summarize the attributes of single- and multiple-site practices and the communities where they were located. Key practice and community variables were analyzed. DATA COLLECTION/EXTRACTION METHODS: The questionnaires were coded and entered into a digital database with the tabulated community data. Responses of individual physicians were grouped by practice. PRINCIPAL FINDINGS: Multiple-site practices were common. Second-order sites accounted for 23 percent of total appointment capacity and were located in communities with higher than average elderly populations and incomes and lower than average minority populations. CONCLUSIONS: Analysis of multiple-site practices is important for the accurate assessment of medical service availability. Further research is needed to document the functioning of multiple-site practices across other specialties and geographic areas. PMID- 8407341 TI - Uncompensated care provided by private practice physicians in Florida. PMID- 8407342 TI - New evidence disproving fears that intramuscular vitamin K might be linked to childhood cancer. PMID- 8407343 TI - Protect free speech and the right to hear. PMID- 8407344 TI - PREP. Almost there. PMID- 8407345 TI - Review teams threaten local care. AB - The six specialty review teams set up to look at the provision of services across London have reinforced the move to close major London hospitals outlined by the Tomlinson review and in the DH report Making London better. The teams suggest major specialties are concentrated on a reduced number of sites. Christina Potrykus examines the implications. PMID- 8407346 TI - Public health. Crossing the borders. AB - Can public health survive the shift in community health service delivery to general practice? Who will pay for it? And who will do it? These were some of the questions debated by delegates at the Public Health Alliance conference 'Making New Friends: the public health challenge in primary care', held in Birmingham on 6 July. Cath Jackson reports from two key workshops. PMID- 8407347 TI - Discipline. What can a parent do? AB - If we agree that a smack is not the answer, what can a parent do to discipline a child? Christine Bidmead describes the Veritas parenting programme, which health visitors can use to help parents towards a non-punitive method of discipline. PMID- 8407349 TI - Reflections on first parent visiting. PMID- 8407348 TI - Group child health assessments. AB - 'Health for all children' emphasises health promotion and parental involvement in child health surveillance checks. Heavy caseloads and the need to incorporate these principles in their practice prompted an Oxfordshire team of health visitors to introduce group health assessments. Here they describe the new approach and the response from parents. PMID- 8407351 TI - School health. Fax of life. PMID- 8407350 TI - Tackling language delay: a groupwork approach. AB - Communication, Articulation and Terrific fun adds up to Speech. Jacqueline Cox and Sylvia Hill describe CATS, a speech and language group for language-delayed children run by health visitors. PMID- 8407352 TI - The health, safety and welfare of travellers. PMID- 8407353 TI - School health. Practical solutions to bullying. PMID- 8407354 TI - Care in the community. PMID- 8407355 TI - Study of a contact between two chromosomally monomorphic races of Sorex araneus L. (common shrew). AB - One hundred and twelve specimens of the common shrew from 21 populations in NE Poland were studied from an area of contact between chromosomal race II (metacentric chromosomes: hi, ko, gm, np) and race VII (ki, hn, gr, mp). No direct contact was found between the races in the study area; there were no mixed or hybrid populations. The shortest distance between populations of the two races was 1.2-1.6 km. Possible explanations for this pattern of the races' disjunction are discussed. PMID- 8407356 TI - The adult component of selection in Drosophila melanogaster: some aspects of early-remating activity of females. AB - As an important factor of the adult component of selection, mating behaviour was studied in Drosophila melanogaster, with emphasis on non-virgin females. We found that 30-50 per cent of the females in a laboratory population will remate within 6 h of first mating under no-choice conditions. This high percentage of early rematings was not due to the continuous confinement of the females with males but indicated a rapid return of receptivity of a significant proportion of the females. Remating behaviour was significantly influenced by both the genotype of the female and the genotype of her two successive partners. Age of females was only important insofar as it concerned young, 1 or 2-day old, females. These females showed less remating than older females. Willingness to remate was also affected by the number of sperm stored. Females that had been inseminated by less fertile males, i.e. males that had already mated two or three times, showed higher remating percentages than females inseminated by more fertile males. Notwithstanding this sperm effect, females were estimated to remate approximately every second day. It is suggested that a high frequency of remating and the resulting sperm competition are significant components of Drosophila life history. PMID- 8407357 TI - Diapause in Drosophila melanogaster females: a genetic analysis. AB - Female Drosophila melanogaster exhibit ovarian diapause at low temperatures and short day lengths. We found that D. melanogaster isofemale lines from Windsor (Ontario, Canada) had a significantly higher percentage of females in diapause than did those from Cartersville (Georgia, U.S.A.). To investigate the heredity of this trait, we performed a 16-reciprocal cross analysis using two extreme isofemale lines called W and C. We found that diapause in D. melanogaster is inherited as a simple autosomal recessive trait with the C response (less flies in diapause) completely dominant to the W one. Maternal and cytoplasmic factors did not affect differences in diapause in these lines. The result of our genetic analysis of diapause in D. melanogaster opens may avenues for the genetic dissection of this ecologically relevant trait. PMID- 8407358 TI - Integrins: cell adhesives and modulators of cell function. AB - Integrins encompass a family of cell-surface molecules which play a crucial role in cell-cell and cell-extracellular matrix interaction. Of these heterodimeric transmembrane glycoproteins (consisting of an alpha and beta chain) as yet at least 20 different types have been described, all with a different pattern of reactivity with extracellular matrix components. In this review the cell and tissue distribution of the integrins is discussed, with special emphasis on immunohistochemical localization of the beta 1 integrins and the alpha 6 beta 4 integrin. The beta 1 integrins comprise a subfamily in which eight alpha chains combine with one beta (the beta 1) chain. The alpha 2 beta 1, alpha 3 beta 1 and alpha 6 beta 1 and the alpha 6 beta 4 integrins are expressed on a wide variety of epithelia on the basolateral surface or exclusively on the basal surface facing the basement membrane (e.g. alpha 6 beta 1 and alpha 6 beta 4). Leucocyte integrins, which share a common beta 2 chain, occur almost exclusively on white blood cells and their precursors. The vitronectin receptors, which share a common alpha v chain, occur in a wide variety of cell types. Integrins play a major role in the interaction of the cell with the extracellular matrix in order to create and maintain tissue architecture. It has become clear, however, that through integrin-ligand interaction cell function is also modulated. Furthermore, in pathological conditions integrins play a role of some significance. Integrins mediate leucocyte traffic in developing inflammatory processes and function in neoplastic growth when it comes to invasion and metastasis. PMID- 8407359 TI - Superoxide formation preceding flight muscle histolysis in Solenopsis: fine structural cytochemistry and biochemistry. AB - In Solenopsis spp., muscle histolysis or breakdown is a normal process in females and is initiated in the flight muscles only immediately after a mating flight. Information regarding the presence of the oxyradical scavenging enzyme superoxide dismutase (SOD) and the formation of the radical oxygen intermediate superoxide (SO) during the early stages of flight muscle histolysis in this insect was investigated. In normal fibrillar flight muscles from control animals, SOD was immunolocalized to vesicular and tubular components of the sarcotubular system. Lanthanum tracer studies indicated that some of these SOD-positive structures might be tubulovesicles continuous with the extracellular space. Following the injection of virgin alates with experimental haemolymph obtained from artificially inseminated females, the membrane delimited elements of the sarcotubular system became increasingly swollen and dilated with time (from 60 to 120 minutes postinjection) with a concomitant decrease in SOD activity and an increase in oxyradical formation. Many similar vesicles were lanthanum-positive. SO was not seen in the sarcoplasmic vesicles and tubules of control insects. The biochemical quantification of SO release over a 2-hour period showed a marked increase in oxyradical formation following treatment with the experimental haemolymph in comparison to control insects. Also, the addition of superoxide dismutase depressed SO formation under these conditions. Despite the histochemical and biochemical changes seen in the muscles of experimental insects, by 2 hours post-treatment there was no evidence of muscle necrosis. From these studies on flight muscle histolysis/necrosis in Solenopsis it appears that the formation of oxyradicals might represent an early event in myopathogenesis and subsequent tissue involution. The generation of SO is more than likely to be associated with alterations in the normal structure, biochemistry and permeability of the biomembranes which delimit the sarcotubular system. PMID- 8407360 TI - Biochemical verification of quantitative histochemical analysis of succinate dehydrogenase activity in skeletal muscle fibres. AB - The purpose of this investigation was to examine critically the validity of a computerized quantitative microphotometric histochemical technique for the determination of succinate dehydrogenase (SDH) activity in skeletal muscle fibres. Sections from the anterior costal diaphragm were removed from Fischer-344 rats (n = 12) and assayed histochemically to determine SDH activity. The SDH activity in individual muscle fibres was computed using a computerized microphotometric histochemical technique which involves measurement of the optical density of deposited diformazan derived from nitroblue tetrazolium within the fibres. To validate the histochemical technique, whole muscle SDH activities were calculated from the histochemical procedure and were compared to SDH activities determined from whole muscle homogenates via a standard quantitative biochemical assay. The mean within-day variability of the computerized microphotometric histochemical technique of determining SDH activity was 6% (range = 0.5-10.9%) for an area containing approximately 50 fibres and 6.1% (range = 1.05-14.9%) for an individual muscle fibre. Similarly, the mean between day variability of the microphotometric histochemical technique of determining SDH activity was 5.9% (range = 2.6-13.9%) for an area containing approximately 50 fibres and 6.6% (range = 2.2-13.9%) for an individual muscle fibre. The inter class correlation coefficient between biochemically determined SDH activity and histochemically determined SDH activity was r = 0.83 (p < 0.05). Collectively, these data demonstrate that the quantitative histochemical technique of Blanco et al. (1988) is both valid and reliable in the determination of SDH activity in skeletal muscle fibres. PMID- 8407361 TI - Potassium-dependent p-nitrophenyl phosphatase and carbonic anhydrase reactivities suggest that lymphoid follicles in the large intestine of lambs are lined with a uniform type of epithelial cell distinct from the absorptive epithelium. AB - The epithelium covering the large intestinal lymphoid follicles in fetal and postnatal lambs was examined for potassium-dependent p-nitrophenyl-phosphatase (K(+)-NPPase), carbonic anhydrase, magnesium-dependent adenosine triphosphatase (Mg(2+)-ATPase) and acid phosphatase. Reactivities for these enzymes indicated a homogenous population of cells in the follicle-associated epithelium (FAE), distinct from the absorptive epithelium. There were essentially no differences in the enzyme reactivities of the large intestinal FAE between fetuses in late gestation and postnatal lambs. The FAE showed a weak reaction for K(+)-NPPase and a variable staining for Mg(2+)-ATPase and acid phosphatase. In contrast, the adjacent absorptive epithelium demonstrated strong reactions for these enzymes. Carbonic anhydrase gave a strong reaction at the luminal and apparent basolateral cell borders of the large intestinal FAE. This distribution of reactivity for carbonic anhydrase resembled that found in the ileal FAE. In absorptive epithelial cells, only the luminal cell border reacted strongly for carbonic anhydrase. Serial sections of large intestinal tissue showed a variation in the basolateral staining of FAE from one section to the next, a finding which suggested that the reaction may be associated with transcytosis. The lymphoid follicles and domes of the large intestine showed a variable granular pattern of carbonic anhydrase staining, which also suggested a dependence on epithelial transcytosis. PMID- 8407363 TI - Advantages of histochemistry for the study of cell biology. AB - Bridging between the two bodies of cell science, histochemistry has brought together knowledge of morphological structures and biochemical constituents and provided insight into the function of one or the other. Where it has localized an enzyme, hormone or other entity of known biological activity to a cell type, histochemistry has contributed insight into the cell's function. By detecting heterogeneity in the content of an enzyme or glycoconjugate within a presumed uniform population of cells, the histochemical approach has differentiated among these cells subtypes with demonstrated or presumed differences in function. On the other hand, in instances where it has located an isozyme, antigen, glycoconjugate or other entity of uncertain significance in a cell or organelle of known function, histochemistry has suggested a possible role for the constituent related to that of the structure. Histochemical examination provides the observer with a different view of body tissues and their composition from that obtained by strictly morphological or chemical techniques. Information acquired through this advantage is cited wherein an immunohistochemical method disclosed previously undiscovered neural organs and carbohydrate histochemistry detected previously unrecognized glycoconjugates. PMID- 8407362 TI - An immunohistochemical study examining the role of collagen type VI in the rodent periodontal ligament. AB - The distribution of collagen types I and VI has been examined in the periodontal ligaments of rat incisor and molar teeth using cryosections and immunohistochemical staining procedures. The stain for collagen type I was uniform in all the ligaments examined. Uniform staining for collagen type VI was evident only in the ligament of the fully erupted molars, and the stain was absent from the mid-zone of the ligaments of the erupting molars and incisors. The staining pattern of the collagen type VI antibodies is consistent with previous reports which have suggested that the removal of collagen type VI precedes the degradation of major banded collagen fibrils in remodelling connective tissues. The removal of collagen type VI from the mid-zone of the incisor ligament may precede the remodelling events which facilitate tooth eruption. PMID- 8407364 TI - Expression of argyrophilic nucleolar organizer regions (AgNORs) can be used to assess cellular proliferation as shown in rat thymic sections. AB - Silver-stained nucleolar organizer regions (AgNORs) were studied in thymic sections from 4- and 30-day-old rats. By direct examination under the light microscope cells with a low or high content of AgNORs (type I and type II cells, respectively) were identified and their relative numbers calculated. The mean area of AgNORs per cell was calculated for each type of cell and age group. Additionally, the proportion of cells labelled with bromodeoxyuridine was calculated in sections from the same animals. Visual identification of type I and type II cells was confirmed by a significant (p < 0.01) difference in the mean AgNOR area in both types of cell. Both the proportion of cells with a high expression of AgNORs (type II cells) and that of bromodeoxyuridine-labelled cells were significantly greater (p < 0.01) in 4-day-old rats than in 30-day-old rats. A significant correlation was found between both variables (R2 = 0.45; p = 0.002), the relation being best between both variables and age (R2 = 0.91; p = < 0.001). These data offer support for an easy interpretation of the AgNOR reaction in which the proportion of cells with a high expression of AgNORs can be used as an index of proliferative activity in a tissue sample. PMID- 8407365 TI - Dystrophin-related protein, utrophin, in normal and dystrophic human fetal skeletal muscle. AB - Dystrophin is the product of the Duchenne muscular dystrophy (DMD) gene. Dystrophin-related protein (utrophin), an autosomal homologue of dystrophin, was studied in skeletal muscle from normal fetuses aged 9-26 weeks and one stillbirth of 41 weeks' gestation, and compared with low- and high-risk DMD fetuses aged 9 20 weeks. Utrophin was present at the sarcolemma from before 9 weeks' gestation, although there was variability in intensity both within and between myotubes. Sarcolemmal immunolabelling became more uniform, and levels of utrophin increased to a maximum at approximately 17-18 weeks. Levels then declined, until by 26 weeks sarcolemmal labelling was negligible and levels were similar to adult control muscle. By 41 weeks there was virtually no sarcolemmal labelling, although immunolabelling of capillaries was bright. Similar results were obtained with normal and DMD fetal muscle. Utrophin is therefore expressed in the presence and absence of dystrophin and down-regulated before birth in normal fetal muscle fibres. Samples were not available to determine whether or when, utrophin levels decline in DMD fetal muscle. On Western blots, utrophin was shown to have a smaller relative molecular mass than adult dystrophin, but similar to the fetal isoform. Blood vessels were brightly immunolabelled at all ages, although utrophin immunolabelling of peripheral nerves increased with gestational age. PMID- 8407366 TI - Presence of lysosomal enzymes in the normal glomerular basement membrane matrix. AB - The question posed in the present study was: are there hydrolytic enzymes, including proteases, present in the extracellular matrix of the glomerular basement membrane? If these enzymes are present they may play a role in the catabolism of the glomerular basement membrane (GBM) and removal of macromolecular debris resulting from ultrafiltration. Enzymes, acid phosphatase- the marker for lysosomal enzymes--beta-galactosidase, beta-glucuronidase and acid protease (using albumin as substrate) were biochemically assayed in purified basement membrane preparations. It was found that all enzymes were present in significant amounts in the basement membrane. Compared to other enzymes, acid protease activity was present in much higher amounts. The pH optima of these enzymes were variable but all had significant activity at neutral pH. A method was developed to localize the marker enzyme, acid phosphatase, ultrastructurally in the basement membrane in order to substantiate the biochemical findings. Activity was shown by the presence of dense deposits of lead phosphate. Staining for acid phosphatase could also be shown on isolated, purified basement membrane. The demonstration of acid hydrolases in the GBM matrix argues for their role in (i) the extracellular turnover of basement membrane macromolecules, and (ii) clearance of debris of ultrafiltration which tend to clog the membrane pores. PMID- 8407367 TI - Reflection contrast microscopy of ultrathin sections in immunocytochemical localization studies: a versatile technique bridging electron microscopy with light microscopy. AB - Reflection contrast microscopy (RCM) of ultrathin sections was recently introduced as a sensitive technique for visualization with enhanced definition in immunogold histochemistry. Experience of using RCM as a major tool in immunocytochemical research in different fields is summarized, e.g. oncology, nephrology and embryology. The sensitive visualization of immunocytochemical labels, gold particles or peroxidase-diaminobenzidine deposits in or on ultrathin sections, by RCM instead of electron microscopy is demonstrated. RCM of ultrathin sections is an adequate light microscopical alternative for immunoelectron microscopy, since an overview of both label and tissue is obtained with a high image definition and high contrast of label. In the studies presented, RCM is shown to provide a better gradation in staining intensity and staining pattern than other light microscopical methods. Moreover, a precise localization of multiple labels is obtained with this method. Besides the applications shown, ultrathin section visualization by RCM is very useful for correlative light- and electron microscopical studies of fine structures. Commercially available fluorescence microscopes can be adapted for proper RCM functioning; an adaptation scheme and list of microscopes tested is provided. PMID- 8407368 TI - Differential expression of drug metabolizing enzymes in primary and secondary liver neoplasm: immunohistochemical characterization of cytochrome P4503A and glutathione-S-transferase. AB - The question whether expression of drug metabolizing enzymes in human liver is altered by liver neoplasm remains controversial; however, the ability or unability of tumour cells to metabolize certain drugs may be important for developing therapeutic strategies. We therefore investigated the abundance and localization of two classes of drug metabolizing enzymes [cytochrome P4503A (CYP3A) and pi-type glutathione-S-transferase] by means of immunohistochemistry (standard ABC technique) in patients with hepatocellular carcinoma (HCC, n = 16) and with liver metastasis from adenocarcinoma (n = 53) in comparison to normal controls (n = 5). The distribution of CYP3A in normal liver samples showed a characteristic pattern of four to five layers of stained hepatocytes surrounding the central vein. Eleven out of 16 cases of HCC showed expression of CYP3A; staining was less intense than in normal liver and zonation was completely lost. In contrast, only 5 out of 53 samples of metastasis stained positively for CYP3A. The difference between primary and secondary neoplasm was statistically significant (chi-square, P < 0.0001). Pi-type glutathione-S-transferase (GST) stained positively in 9 out of 16 HCC and in 48 out of 53 cases of liver metastasis (chi-square, P < 0.01) indicating a higher percentage of immunostaining in liver metastasis. In summary, we observed differences in the abundance and distribution pattern of CYP3A and GST between primary and secondary neoplasma of human liver and in comparison to normal controls. In combination with established methods these data may contribute to the establishment of reliable test systems for distinguishing primary from secondary liver tumours.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407369 TI - Digestion of centromeric DNA from each human metaphase chromosome by the 6 bp restriction enzyme StuI. AB - Although 6 bp-restriction endonucleases infrequently cut DNA, we describe in this paper the banding pattern induced by one of them, StuI (AGGCCT), on fixed human chromosomes. This enzyme is the first 6 bp-restriction endonuclease demonstrated to be able to digest human heterochromatin. It causes the extraction of two families of repeated DNA, the alpha satellite DNA and the 3.4 kb HaeIII family (DYZ1). On the other hand, digestions with StuI and with another two enzymes, HinfI and RsaI, have established the distribution of sequences within the heterochromatic block of chromosome 3. PMID- 8407370 TI - A new approach to the immunocytochemistry of cAMP. Initial characterization of antibodies against acrolein-fixed cAMP. AB - A method is described to couple cyclic adenosine 3',5' monophosphate (cAMP) to a carrier protein by means of acrolein. Antibodies against this conjugate were raised in mice. These antibodies proved to be highly specific for acrolein-fixed cAMP in a gelatin model system. Slices (300 microns in thickness) from rat cerebral cortex were incubated in vitro and the dopaminergic control of adenylate cyclase activity was drug-manipulated. This manipulation was visualized by application of the cAMP-antisera on cryostat sections of the acrolein fixed slices. PMID- 8407371 TI - Immunocytochemical localization of delta 3, delta 2-enoyl-CoA isomerase and NADPH dependent-2,4-dienoyl-CoA reductase in rat kidney. AB - Localization of delta 3, delta 2-enoyl-CoA isomerase (ECI) and NADPH-dependent 2,4-dienoyl-CoA reductase (DCR) in the rat kidney was investigated by immunocytochemical techniques. The kidneys were perfusion-fixed and embedded in Epon or LR White. For light microscopy, semi-thin sections of Epon-embedded materials were stained by the immunoenzyme technique after the epoxy resin was removed by treatment with sodium ethoxide. For electron microscopy, ultra-thin sections of LR White-embedded materials were stained by the protein A-gold technique. By light microscopy, the S1 segment of the proximal tubule was most heavily stained for ECI and DCR whilst S2 and S3 segments showed intermediate staining. A weak staining reaction was observed in the distal tubule and the medullary collecting tubule. In the cortical collecting tubule, heavily stained cells were present between weakly stained cells. By electron microscopy, gold particles showing the antigenic sites for ECI were confined mainly to the mitochondria, but few particles were observed in the peroxisomes. Gold labeling for DCR was localized both in the mitochondria and the peroxisomes. The labeling intensity of the peroxisomes was much higher than that of the mitochondria. The results suggest that metabolism of unsaturated fatty acids occurs mainly in the mitochondria and the peroxisomes of the proximal tubule in the kidney. PMID- 8407372 TI - A new enzyme-linked immunosorbent assay (ELISA) for studying immunocytochemical procedures using an antiserum produced against spermidine as a model. AB - Antiserum was produced in rabbits against the polyamine spermidine (Spd) conjugated to bovine serum albumin (BSA). The reactivity of the serum to Spd and a variety of structurally related compounds was quantified by a new immunocytochemical model system incorporating an enzyme-linked immunosorbent assay (ELISA) binding test. This is based on the principle of coupling these compounds to the wells of microtiter plate activated with poly-L-lysine and glutaraldehyde and incubating the wells by the indirect immunoperoxidase method. The antiserum showed a 25% cross reaction with spermine (Spm), putrescine (Put), and cadaverine (Cad), and a 1% cross reaction with 1,3-diaminopropane (Dap), but no cross reaction with monoacetyl polyamines and amino acids. The antibody binding was inhibited most effectively by absorption of the antiserum with N1 acetylspermidine and Spd in the ELISA inhibition test. Also, immunoblot analysis of the antiserum with nitrocellulose paper gave completely identical results to the ELISA binding tests. Spd-like immunoreactivities in human melanoma BD and neuroblastoma IMR 32 cell lines are presented as examples of the staining pattern obtained with the antiserum. Absorption of the serum with N1-acetylspermidine and Spd was demonstrated to abolish the immunostaining reaction. The immunohistochemical model is simple: amines and amino acids are bound in the same way as in aldehyde-fixed tissues and, in comparison to immunoblot analysis, the immunoreactivity can be more easily and accurately quantified by assay with the antibody. The model should prove useful in assessing the specificity of other antisera. PMID- 8407373 TI - Immunocytochemical localization of prostaglandin endoperoxide synthase in the bovine intestine. AB - The localization of prostaglandin (PG) endoperoxide synthase in bovine intestine was examined immunocytochemically with polyclonal antibody raised against PG endoperoxide synthase purified from bovine seminal glands. The most intense positive staining reaction for the enzyme was present in mast cells. Mast cells were found to be widely distributed in the intestinal wall, and were particularly numerous in the lamina propria. Most of the mast cells in the lamina propria of the intestinal villi were elongated and oriented with their long axis parallel to the plane of the absorptive epithelium. In whole mount preparations of jejunal villi, mast cells were seen to form a two-dimensional network in the lamina propria. In addition to mast cells, smooth muscle cells of the inner circular muscle layer and muscularis mucosae, nerve cells and fibers, endothelial cells of arterioles, and serosal epithelial cells also showed faint to moderate staining for the enzyme. These results suggested that mast cells are the major source of PGs in the bovine intestinal wall. The characteristic arrangement of mast cells in the intestinal villi may be related to their functions in this portion of the bovine intestine. PMID- 8407374 TI - [Basic principles of lithotripsy]. PMID- 8407375 TI - [German ENT Congress in Munster. Diagnosis and therapy in the anterior cranial base]. PMID- 8407376 TI - [Middle ear surgery]. PMID- 8407377 TI - [Laser surgery]. PMID- 8407378 TI - [Indications for psychotherapeutic treatment of functional dysphonias]. AB - Functional voice disorders require adequate psychiatric screening and treatment due to the complex etiologies of the psychic elements involved. While the diagnosis of dysphonia increasingly takes into account various emotional factors, considerations of indications for psychotherapy and for differing forms of psychotherapy are missing in the literature. To fill this gap, the aims and principles of a number of psychoanalytically based therapeutic methods are presented. Criteria for general indications for treatment and with respect to patients with functional voice disorders are outlined. Case reports of patients with functional voice disorders illustrate individual methods for diagnosis and treatment. Finally, the necessity, for the development of general criteria for differential indications for psychotherapy in functional voice disorders is pointed out. PMID- 8407379 TI - [Functional results and survival probability of tumor patients after reconstruction of the mouth cavity and oropharynx using a microvascular radial forearm flap]. AB - In tumor surgery of the oral cavity and oropharynx function and quality of life should be maintained and large resection margins to avoid recurrences. For these purposes the radial forearm flap has proved to be a versatile tissue transfer since it is thin and pliable and has a long vascular pedicle. From 1987 to 1991 we used the radial forearm flap in performing 70 reconstructions of the oral cavity and oropharynx after resection of squamous cell carcinomas. Forty-six patients had carcinomas of the oropharynx, while 24 patients had carcinomas of the oral cavity. The indications for these reconstructions were tumors of the oral cavity and oropharynx greater than T2 which after resection were not suitable for primary closure of the defect. Thirty-seven patients died during the follow-up period, with 36% dying within the first 2 years after operation. Fifty five percent of these patients died of recurrences, 17% of metastases and 11% of intercurrent diseases. In 17% of cases the cause of death was unknown. The 2-year survival probability was 52% (Kaplan Meier). Our results show that reconstructions with the radial forearm flap do not improve survival rates when compared to the general survival rate in these cases despite a possibly larger resection margin allowing a more radical tumor resection. Thirty-one of the 33 patients still alive underwent following examinations. Forty-six percent of the patients with tumors of the oropharynx and 57% of the patients with tumors of the oral cavity had severe difficulties in swallowing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407380 TI - [How badly does the "normal-hearing" young man of 1992 hear in the high frequency range?]. AB - In the course of a clinical study we examined young men believed to have normal hearing. Volunteers with a positive history for noise deafness were excluded from further study. Normal hearing was verified by pure-tone audiometry if the hearing loss did not exceed 15 dB for each test tone between 250 Hz and 8000 Hz. As this limit is far below normative threshold values (DIN ISO 7029) we expected to find a large number of otological normal subjects. Using pure-tone audiometry, ABR and noise exposure, we found that 65% of the young male volunteers (mean age 25 years) did not meet the criteria for audiologic normality. The pure-tone thresholds for these subjects were comparable to the expected levels of their fathers' generation. PMID- 8407381 TI - [Electrohydraulic intracorporeal lithotripsy of salivary calculi. In vitro and animal experiment studies]. AB - BACKGROUND: Extracorporeal lithotripsy is now used routinely for the treatment of salivary duct stones. The question arose whether electrohydraulic intracorporeal lithotripsy, which is applied in urology and gastroenterology, might also be useful in the treatment of this disease. Before its possible clinical application the influence of electrohydraulic intracorporeal shock waves on salivary stones in vitro and any influence on the tissue in the head and neck region (in vivo) had to be investigated. MATERIALS AND METHODS: In vitro experiments. Fifty-eight salivary stones and 11 extirpated human submandibular glands were treated by three different electrohydraulic devices. Animal experiments. Electrohydraulic shock waves were applied to the dilated Stensen's duct and other tissues (muscle, parotid gland, facial nerve) of six rabbits. RESULTS: Of 58 salivary stones, 53 (91%) were fragmented, 39 (67%) with a remaining size of less than 1.5 mm and 14 (24%) more than 1.5 mm. In 5 cases (9%) no effects were seen at all. Fragmentation occurred independent of the mineralogical components of the stone and independent of the different lithotriptors used. The smaller the probe diameter and the larger the stone, the more shock waves were needed to achieve fragmentation. With the smaller probes the stones could not be fragmented completely. In human submandibular glands, extensive tissue lesions could be evidenced macroscopically and histologically after application of electrohydraulic shock waves in vitro. Application of electrohydraulic shock waves to the dilated parotid gland duct of rabbits led to perforations of the duct after 1-5 single pulses. Lesions of nerves and blood vessels could also be observed within the duct environment. This occurred with all of the different electrohydraulic devices, probe diameters and intensities used. In our opinion the damage produced is probably the result of both the direct effect of the plasma as well as the resultant stress wave. CONCLUSION: In view of the severe damage caused to different tissues, use of the described method for the treatment of stones in the narrow human salivary ducts should be greatly restricted. PMID- 8407382 TI - [Myoepithelial carcinoma of the nasopharynx. Case report and review of the literature]. AB - Minor salivary gland neoplasms are extremely rare in the nasopharynx. We herein report a case of a 68-year-old man with a rapidly expanding myoepithelial carcinoma of the nasopharynx. The classification of this carcinoma as well as the difficulty in effective treatment are discussed A review of the literature is given. PMID- 8407383 TI - [Otorhinolaryngologic aspects of diving sports]. AB - ENT disorders are the most common of all medical problems of diving. This review summarizes the specific conditions and ENT diseases in Scuba diving. During compression failure to equalize the pressure of air-filled cavities surrounded by bone deprives the middle ear or sinuses of aeration. Middle ear barotrauma is the most common barotrauma encountered in divers while sinus barotrauma and especially inner ear barotrauma (with rupture of the round or oval window) are less common. Decompression sickness in primarily the result of inert gas bubbles; deafness and vertigo may result if the inner ear is involved. The ENT examination necessary for assessment of diving fitness focuses on the middle and inner ear as well as the nose, sinuses and larynx. A list of ENT contra-indications is presented that mandate temporary or permanent disqualification from diving. PMID- 8407384 TI - Readers offer suggestions for reforms in osteopathic medicine. PMID- 8407386 TI - Stroke requires treatment--stat! PMID- 8407385 TI - Readers offer suggestions for reforms in osteopathic medicine. PMID- 8407387 TI - Comparison of thoracic manipulation with incentive spirometry in preventing postoperative atelectasis. AB - Atelectasis is a preventable complication that often occurs after upper abdominal surgery. In our 1-year randomized, researcher-blinded trial, low-risk cholecystectomy patients were subjected to either the thoracic lymphatic pump (n = 21) or incentive spirometry (n = 21) to prevent atelectasis. The treatment groups were equal with respect to risk factors for atelectasis and deviation of preoperative respiratory parameters (forced vital capacity [FVC] and forced expiratory volume in one second [FEV1]) from the predicted values. Atelectasis occurred in 2 (5%) of 21 patients regardless of whether incentive spirometry or thoracic lymphatic pump treatment was used. Study patients treated with the thoracic lymphatic pump technique had an earlier recovery and quicker return toward preoperative values for FVC and FEV1 than patients treated with incentive spirometry. PMID- 8407388 TI - Standards for Pediatric Immunization Practices: an osteopathic physician's commentary and 'reality check'. AB - The "Standards for Pediatric Immunization Practices," recommended by the National Vaccine Advisory Committee and approved by the US Public Health Service, provides guidance to clinicians as to what the National Vaccine Advisory Committee considers optimal practice. Each of the 18 recommendations reflects the ideal; unfortunately, life in the trenches is less than ideal. Providers must reflect on each point and consider how they might modify their own practices to help us to realize the goal of Healthy People 2000, that is, 90% immunization coverage by the child's second birthday. The Standards are clear. They are reprinted herein from MMWR (April 23, 1993;42[5]). Following each is a response, sort of a reality check, from the author, a practicing clinician and practice administrator. PMID- 8407389 TI - The HIV-infected house officer: residency training issues. AB - Efforts to fully integrate physicians infected with the human immunodeficiency virus (HIV) into residency programs have been complicated by concerns of the potential risks of viral transmission from physician to patient. Despite numerous studies, this potential risk has not been quantified. This article addresses the issue of HIV-infected interns and residents, classifying HIV as a potential disability. The suggested recommendations emphasize routine monitoring and evaluation of professional competence and compliance with proper infection control procedures, as delineated by the Centers for Disease Control and Prevention. The need for hospitalwide guidelines and adequate health and disability insurance are also discussed. PMID- 8407390 TI - Exuberant keloidal formation. AB - Keloids are disfiguring deformities that occur after trauma or wounding of the skin, most commonly among blacks. The case presented here is unusual because of the number of keloids and the massive size of one. The patient underwent full surgical excision together with intraoperative and postoperative steroid injections with good cosmetic results. Various causes of keloids are reviewed as are the histopathologic differences between hypertrophic scars and keloids. Alternative surgical and nonsurgical treatment techniques are examined. PMID- 8407391 TI - 20-year experience in childhood craniopharyngioma. AB - PURPOSE: The management of craniopharyngioma is controversial, and surgery alone is frequently advocated. The purpose of this study was to assess the long-term impact of various treatments in childhood craniopharyngioma. METHODS AND MATERIALS: Sixty-one children < or = 21 years of age at diagnosis were treated for craniopharyngioma at Children's Hospital and the Joint Center for Radiation Therapy in Boston from 1970 to 1990. The median age was 7.5 years (range 10 months-21 years). There were 33 females and 28 males. The median follow-up was 10 years (range 2-20.5 years). Neuroimaging was available for detailed review in 53. Nine children were treated with radiotherapy alone, 15 were treated with surgery alone, and 37 were treated with both surgery and radiotherapy. All patients in the radiotherapy and surgery plus radiotherapy groups were treated with megavoltage radiation with a median dose of 5464 cGy. RESULTS: All nine of the children treated with radiation therapy alone are alive; none have recurred. Nine of the 15 children treated with surgery alone have recurred (p = 0.007 Fisher exact test). Two are alive with disease, and seven are alive without disease after treatment at relapse with radiation therapy, surgery, or both. Seven of the 37 patients treated with surgery plus radiotherapy have recurred. Three of the seven patients are dead of disease, three patients are alive with disease, and one patient is alive without disease after further treatment. The 10-year actuarial overall survival was 91% for all patients. The 10-year actuarial freedom from progression for the surgery group was 31% compared with 100% for patients treated with radiation therapy only (log rank p = 0.01), and 86% for patients treated with surgery plus radiotherapy at diagnosis (p = 0.001). There were two treatment related deaths, both in the surgery plus radiotherapy group. A higher incidence of visual loss and diabetes insipidus was associated with the use of aggressive surgery. The size of the tumor at presentation correlated with an increased risk of recurrence; 5 of 6 patients with tumors > or = 5 cm experienced recurrences while only 6 of 30 recurred when the tumor was < 5 cm. CONCLUSIONS: Overall survival in childhood craniopharyngioma is excellent. However, patients treated with surgery alone have a significantly worse freedom from progression when compared to patients treated with surgery and radiation therapy or radiation therapy alone. PMID- 8407392 TI - Final results of a study of escalating doses of hyperfractionated radiotherapy in brain stem tumors in children: a Pediatric Oncology Group study. AB - PURPOSE: In September 1984, the Pediatric Oncology Group began accrual to a Phase I/II study designed to assess the efficacy and toxicity of sequentially escalated doses of hyperfractionated (twice daily) radiotherapy in children with poor prognosis brain stem tumors. Pediatric Oncology Group Study #8495 closed in June 1990 with a total of 136 patients on study. We report here the outcome of patients treated at the third and final dose level (75.6 Gy), and compare the results to those obtained at the 66 and 70.2 Gy dose levels. METHODS AND MATERIALS: Patients eligible for study were those between 3 and 21 years of age with previously untreated tumors arising in the midbrain, pons or medulla. Histological confirmation of diagnosis was not mandatory provided that the clinical and radiological findings were typical for brain stem glioma. Treatment consisted of radiotherapy delivered to local fields. At the third dose level, fraction sizes of 1.26 Gy were given twice daily, with a minimum interfraction interval of 6 hr to a dose of 75.6 Gy in 60 fractions over 6 weeks. Between 5/89 and 6/90, 41 patients were accrued to the study. Two were excluded from analysis leaving 39 evaluable patients, 21 male and 19 female, whose ages ranged from 3 to 15 years (median 7.5 years). RESULTS: Following treatment, neurological improvement was reported in 30/39 (77%) of the patients. On central review of imaging studies in 29 patients, one patient was found to have had a complete response to radiotherapy, five a partial (> 50% response), and only three had non responding or progressive disease. The median time to disease progression was 7 months; median survival time was 10 months; survival at 1 year was 39.9% (SE 8.3%) and at 2 years, 7% (SE 4.8%). The pattern of failure was local in all patients; in addition six had evidence of leptomeningeal seeding. Morbidity of treatment included an enhanced skin reaction (21%), otitis media and/or externa (26%), and steroid use > 3 months (62%). Intralesional necrosis was a frequent finding (45%) on imaging studies performed at a median time of 6 weeks post treatment. CONCLUSION: The results of treatment in terms of progression-free survival and overall survival are not significantly different (at p = .55 and p = .46, respectively) from those obtained at the two previous dose levels. There is no evidence that higher doses of hyperfractionated radiotherapy given as in this study improve the outlook of patients with poor-risk brain stem gliomas. PMID- 8407393 TI - Iododeoxyuridine (IUdR) combined with radiation in the treatment of malignant glioma: a comparison of short versus long intravenous dose schedules (RTOG 86 12). AB - PURPOSE: To evaluate the toxicity and tumor efficacy of the halopyrimidine IUdR (NSC #39661, IND 22475) as a chemical modifier of radiation response when used in a high dose short time infusion versus the acceptable 4 day infusion. METHODS AND MATERIALS: In August 1987 we initiated a prospective study in patients with newly diagnosed anaplastic astrocytoma and glioblastoma. The study was designed to have a fixed dose of radiation (60.16 Gy = 1.88 Gy in 32 fractions in 6.5 weeks) but varying the dose schedule of IUdR, keeping the total dose between 21 and 24 g/m2. IUdR was delivered in a 96, 48, or 24 hr continuous intravenous infusion per week for 6.5 weeks during radiation treatment. RESULTS: The study was closed for patient accrual on October 1, 1991. Twenty-two patients were treated on the 96 hrs, 32 on the 48 hr and 25 on the 24 hr schedules. The incidence of glioblastoma ranged between 68 and 75% in the three arms. Seventy percent of the patients had a Karnofsky of 80-90% at the onset of treatment. Over 50% of the patient population were under age 55. Drug tolerance was related to the duration of the IUdR infusion. Toxicities were most pronounced in the 96 hr IUdR infusion schedule where 27.4% of the patients reported a grade 3 drug toxicity. No fatal or grade 4 toxicities were observed. More patients on the 24 and 48 hr schedule received at least 80% of the IUdR dose specified per protocol. We did not observe a trend in acute normal tissue radiation reactions in any of the three arms. The median survivals calculated from the Kaplan-Meier plot are 13.4, 10.5, and 11 months, respectively, for the 96, 48, and 24 hr infusions. The Cox Proportional Hazards model showed that any difference in survival can be attributed to histological grade, type of previous surgery and, to some extent, age of the patient. Dose schedule was not a significant predictor of survival, although statistically nonsignificant trend toward longer survival is seen in those patients with glioblastoma treated in the "long" 4 day schedule. CONCLUSION: Overall, our treatment combination, particularly for patients with glioblastoma, has not shown convincing evidence of an improvement in survival. Of interest, however, it is the 2 year survival rate of 68% for patients with anaplastic astrocytoma. In our experience, the administration of IUdR is laborious, time consuming and with bothersome acute gastrointestinal and hematological toxicities. PMID- 8407394 TI - Tolerance of cranial nerves of the cavernous sinus to radiosurgery. AB - PURPOSE: Stereotactic radiosurgery is becoming a more accepted treatment option for benign, deep seated intracranial lesions. However, little is known about the effects of large single fractions of radiation on cranial nerves. This study was undertaken to assess the effect of radiosurgery on the cranial nerves of the cavernous sinus. METHODS AND MATERIALS: We examined the tolerance of cranial nerves (II-VI) following radiosurgery for 62 patients (42/62 with meningiomas) treated for lesions within or near the cavernous sinus. Twenty-nine patients were treated with a modified 6 MV linear accelerator (Joint Center for Radiation Therapy) and 33 were treated with the Gamma Knife (University of Pittsburgh). Three-dimensional treatment plans were retrospectively reviewed and maximum doses were calculated for the cavernous sinus and the optic nerve and chiasm. RESULTS: Median follow-up was 19 months (range 3-49). New cranial neuropathies developed in 12 patients from 3-41 months following radiosurgery. Four of these complications involved injury to the optic system and 8 (3/8 transient) were the result of injury to the sensory or motor nerves of the cavernous sinus. There was no clear relationship between the maximum dose to the cavernous sinus and the development of complications for cranial nerves III-VI over the dose range used (1000-4000 cGy). For the optic apparatus, there was a significantly increased incidence of complications with dose. Four of 17 patients (24%) receiving greater than 800 cGy to any part of the optic apparatus developed visual complications compared with 0/35 who received less than 800 cGy (p = 0.009). CONCLUSION: Radiosurgery using tumor-controlling doses of up to 4000 cGy appears to be a relatively safe technique in treating lesions within or near the sensory and motor nerves (III-VI) of the cavernous sinus. The dose to the optic apparatus should be limited to under 800 cGy. PMID- 8407395 TI - Spinal cord ependymomas: a retrospective analysis of 59 cases. AB - PURPOSE: To determine the outcome of patients with primary spinal ependymoma treated with postoperative radiotherapy and to identify clinical and treatment variables predictive of outcome. METHODS AND MATERIALS: A retrospective chart review was undertaken of 59 spinal ependymoma patients referred to the Princess Margaret Hospital between 1958 and 1987. All patients were treated with radiation therapy to either the site of the primary tumor or the craniospinal axis. There were 23 female and 36 male patients with a median age of 37 years (range: 8-66 years). Median follow-up was 130 months (range: 1-371 months). Clinical variables including age, sex, length of symptoms, functional status, tumor location, and grade as well as treatment variables including extent of surgery, radiation dose and treatment volume were analyzed for influence on outcome. RESULTS: Treatment was well-tolerated with no cases of radiation myelopathy identified. Overall actuarial survival at 5 and 10 years was 83% and 75%, respectively. Eleven patients had recurrent tumor with the 9/11 having a component of their recurrence within the treatment field. Median time to recurrence was 2 years with 9 of the 11 recurrences within the first 3 years. Tumor grade was the only independent variable identified as predictive of outcome. Patients with well differentiated tumors had a 5-year cause-specific survival of 97% compared to 71% for those with intermediate or poorly differentiated tumors (p = 0.005). CONCLUSION: We conclude that postoperative irradiation for patients with spinal ependymomas is associated with a favorable outcome and that tumor grade is a major prognostic factor. Based on results of this series and a review of the literature, specific management recommendations are made for this rare tumor. PMID- 8407396 TI - The management of metastatic spinal cord compression: a radiotherapeutic success ceiling. AB - PURPOSE: In assessing the effectiveness of the management of metastatic spinal cord or cauda equina compression, we performed a retrospective analysis of 70 patients with this complication whom we treated from 1985 to 1989. METHODS AND MATERIALS: The most frequent primary diagnoses in our series were carcinomas of unknown origin and of the breast, lymphoproliferative disease, lung cancer, and prostatic carcinoma. We used the Findlay classification to group all patients according to their pre-therapeutic functional motor status as Grade I (24 patients or 34%), Grade II (27, or 39%) or Grade III (19 or 27%). Treatment consisted of 30-45 Gy of irradiation (using two different schedules) together with high-dose dexamethasone; in only five cases was there surgical intervention. RESULTS: We found that a powerful predictor of response to radiotherapy was the patient's neurologic status (Findlay grade) at the time of diagnosis: 66% of previously ambulatory patients remained so, whereas 30% of non-ambulatory patients and only 16% of paraplegic patients regained the ability to walk. Another important predictor of response was primary tumor histology, with the most favorable responses to radiation therapy having been observed in lymphoproliferative diseases and in breast cancer, but with some response in other radiosensitive malignancies as well. CONCLUSION: The similarity of our results to those of other centers leads us to conclude that a radiotherapeutic success ceiling of 80% may have been reached for Findlay Grade I patients with metastatic spinal cord compression. In view of this, we suggest that future therapeutic endeavour would be best directed toward early diagnosis of the condition. PMID- 8407397 TI - Neutron versus photon irradiation for unresectable salivary gland tumors: final report of an RTOG-MRC randomized clinical trial. Radiation Therapy Oncology Group. Medical Research Council. AB - PURPOSE: To compare the efficacy of fast neutron radiotherapy versus conventional photon and/or electron radiotherapy for unresectable, malignant salivary gland tumors a randomized clinical trial comparing was sponsored by the Radiation Therapy Oncology Group in the United States and the Medical Research Council in Great Britain. METHODS AND MATERIALS: Eligibility criteria included either inoperable primary or recurrent major or minor salivary gland tumors. Patients were stratified by surgical status (primary vs. recurrent), tumor size (less than or greater than 5 cm), and histology (squamous or malignant mixed versus other). After a total of 32 patients were entered onto this study, it appeared that the group receiving fast neutron radiotherapy had a significantly improved local/regional control rate and also a borderline improvement in survival and the study was stopped earlier than planned for ethical reasons. Twenty-five patients were study-eligible and analyzable. RESULTS: Ten-year follow-up data for this study is presented. On an actuarial basis, there continues to be a statistically significant p = 0.009) but there is no improvement in overall survival (15% vs. 25%, p = n.s.). Patterns of failure are analyzed and it is shown that distant metastases account for the majority of failures on the neutron arm and local/regional failures account for the majority of failures on the photon arm. Long-term, treatment-related morbidity is analyzed and while the incidence of morbidity graded "severe" was greater on the neutron arm, there was no significant difference in "life-threatening" complications. This work is placed in the context of other series of malignant salivary gland tumors treated with definitive radiotherapy. CONCLUSIONS: Fast neutron radiotherapy appears to be the treatment-of-choice for patients with inoperable primary of recurrent malignant salivary gland tumors. PMID- 8407398 TI - Place of Iridium 192 implantation in definitive irradiation of faucial arch squamous cell carcinomas. AB - PURPOSE: We have reviewed the results of 165 T1 and T2 squamous cell carcinomas of the faucial arch treated by definitive irradiation including or not Iridium 192 brachytherapy to ascertain whether a significant relationship existed between Iridium implantation, local control, complications, and survival. METHODS AND MATERIALS: From March 1971 to November 1990, 58 T1 and 107 T2 (NO: 107/165; N1: 30/165; N2: 9/165; N3: 19/165) biopsy proven squamous cell carcinomas of the tonsillar region (104/165) and the soft palate and uvula (61/165) were treated in Henri Mondor Hospital by definitive irradiation with curative intent. From 1971 to 1981 (period 1), only guide gutter technique was available, so that implants were reserved for small tumors: patients were either managed by definitive telecobaltherapy to tumor site and neck node areas (Group 1; n = 48; mean dose: 70 Gy; confidence interval: +/- 5.5; 5 fractions of 1.8 Gy per week) or by exclusive Iridium implant (Group 2; n = 11; all T1NO; 64 Gy +/- 4.8) or by a combination of external beam radiation therapy to tumor site and neck nodes areas and Iridium implant (Group 3; n = 40). In 1981 (Period 2), a new plastic tube technique, which enables implantation of larger areas, was introduced in the department and all patients (Group 4; n = 66) were then managed by external radiation therapy (Group 3 + 4: 47 Gy +/- 4.3) followed by an Iridium implant (31 Gy +/- 10.5). Clinically positive neck nodes either received additional external dose with electrons or were excised. RESULTS: Overall 5-year survival (Kaplan Meier) was 21%, 50.5%, and 60% in groups 1, 2, and 3 + 4, respectively (p < 0.001, log rank). Five-year local control was 58%, 100%, and 91%, respectively (p < 0.001). Five-year necrosis rate was 4.5%, 20.5% and 18%, respectively (N.S.). Comparison of results between the two periods of the study (Group 1 + 2 + 3 vs. group 4) show that these two groups are statistically comparable according to site and size of tumor and N status and that both local control (77% vs. 94% at 5 years; p < 0.01) and disease-free survival (56% vs. 71%; p = 0.03) were improved after 1980, while there was a trend to an increase in overall survival (42% vs. 53% at 5 years; p = 0.08); nodal control (86% vs. 95% at 5 years), and necrosis rate (11% vs. 20% at 5 years) were not modified. Multivariate analysis showed that both local control (p < 0.0001) and overall survival (p < 0.0001) were improved when tumor was implanted. CONCLUSION: We recommend then to treat T1 and T2 squamous cell carcinomas of the faucial arch by external radiation therapy to tumor site and neck areas (45 Gy/25 fractions/5 weeks) followed by a 30 Gy Iridium implant and, for patients with clinically positive nodes, either a further 25-30 Gy electron beam irradiation to the nodes or neck node dissection. PMID- 8407399 TI - Long-term results of a prospective randomized trial of adjuvant brachytherapy in the management of completely resected soft tissue sarcomas of the extremity and superficial trunk. AB - PURPOSE: A prospective randomized trial evaluating adjuvant brachytherapy for soft tissue sarcomas was conducted between July 1982 and July 1987. METHODS AND MATERIALS: One hundred twenty-six patients with soft tissue sarcoma of the extremity or superficial trunk (STS) underwent grossly complete resection with limb-sparing surgery. Intra-operatively, patients were randomized to receive either adjuvant brachytherapy (BRT) or no further therapy (no BRT). BRT consisted of an Iridium-192 implant which delivered 4200-4500 cGy over 4-6 days. Total hospital stay for combined surgery and BRT was 10-14 days. Patients in each group were well matched with respect to age, sex, site, tumor size, depth, histologic type, and grade. Median follow-up is 66.5 months for all living patients. RESULTS: At 5 years, local control was 82% in the BRT group vs. 67% in the no BRT group (p = .049). When analyzed by histologic grade, high grade tumors had local control of 90% with BRT vs. 65% with no BRT (p = .013). There was no difference in local control in the low grade patients in either arm. At 5 years, the proportion free of distant metastases was approximately 76% in both arms, with no difference between BRT and no BRT. When analyzed by grade, high grade patients had a similar proportion free of distant metastasis in the BRT vs. no BRT arms despite improved local control in those receiving BRT. Similarly, the disease specific survival for all patients at 5 years was 81% for BRT vs. 80% for no BRT (p = NS). When analyzed by grade, and treatment (BRT vs. no BRT), 5-year disease specific survival for high grade was the same in both groups. CONCLUSIONS: Adjuvant BRT significantly improves local control in completely resected STS. This improvement is confined to patients with high grade histology. BRT provides excellent local control in this group, and significantly shortens the treatment time when compared to adjuvant external beam radiation. However, the improved local control does not translate into either decreased distant metastasis or increased disease-specific survival. PMID- 8407400 TI - Radiation therapy for squamous cell carcinoma of the nasal vestibule. AB - PURPOSE: A retrospective review of carcinomas of the nasal vestibule seen at the Queensland Radium Institute over a 30-year period was undertaken. METHODS AND MATERIALS: Twenty-nine patients with squamous cell carcinoma of the nasal vestibule who were treated with curative intent between 1960 and 1989 were analyzed. The 5-year actuarial survival and disease-free survival were 64% and 61%, respectively. RESULTS: Seven patients were treated with combined modality treatment using surgery and post-operative radiotherapy and 22 were treated with radiation alone. The ultimate local control for those treated with combined modality treatment was 66% and for those treated with radiation alone was 68%. The 5-year actuarial survival was 57% and 67%, respectively. CONCLUSION: In view of the similar local control rates and 5-year survival in the two groups, we suggest that combined modality treatment may not be warranted in the initial treatment of patients with these tumors. Radiation treatment alone with salvage surgery would appear to be the treatment of choice. PMID- 8407401 TI - Results of high-dose thoracic irradiation incorporating beam's eye view display in non-small cell lung cancer: a retrospective multivariate analysis. AB - PURPOSE: To review the University of Michigan clinical experience in nonsmall cell lung cancer using high-dose thoracic irradiation (> or = 60 Gy) so that a starting dose for our prospective dose-escalation study could be determined. METHODS AND MATERIALS: Eighty-eight consecutive patients diagnosed with medically inoperable or locally advanced, unresectable nonsmall cell lung cancer were identified who were treated with thoracic irradiation alone to a minimum total dose of 60 Gy (uncorrected for lung density). All patients except four (95%) underwent computed tomography scanning for treatment planning that included beam's eye view display for tumor and critical structure localization. All patients were treated with standard fractionation in a continuous course to uncorrected total doses ranging from 60 to 74 Gy (median, 67.6 Gy). RESULTS: The median follow-up exceeds 24 months for all surviving patients (range, 12 to 78 months). The median survival time was 15 months, and the 2- and 3-year overall actuarial survival rates were 37% and 15%, respectively. Survival was significantly different between stage of disease (p = .004) and N-stage (p = .002) by univariate analysis. In a multivariate analysis, stage becomes the only characteristic significantly associated with outcome. The median time to local progression for 86 evaluable patients was 29 months. Stage (p = .0003), T-stage (p = .0095) and N-stage (p = .027) were significantly different with respect to local progression-free survival by univariate analysis. However, only stage was prognostic for local progression-free survival by multivariate analysis. There was no difference between large volume treatment (inclusion of the contralateral hilar and supraclavicular lymph nodes) and small volume treatment (exclusion of these elective nodal sites) with respect to local progression-free survival (p = .507) or survival (p = .520). With regard to dose, there was no significant difference between patients who received > 67.6 Gy and patients who received < or = 67.6 Gy with respect to local progression-free survival (p = .094) or survival (p = .142). Within the Stage III subgroup, local progression-free survival (p = .018) and survival (p = .061) were longer favoring the high-dose group of patients. Despite these doses, disease progression within the irradiated field was the predominant first site of treatment failure. CONCLUSION: This retrospective study has shown that it is feasible to deliver uncorrected tumor doses as high as 70 Gy using standard fractionation in NSCLC with acceptable morbidity. Local control remains a significant problem. These data indicate justification for a starting dose in a prospective radiation dose-escalation study. PMID- 8407402 TI - Therapeutic factors influencing the cosmetic outcome and late complications in the conservative management of early breast cancer. AB - PURPOSE: To investigate the impact of clinical and physical parameters on the cosmetic result and complications, in cancer breast treated with conservative surgery and definitive irradiation. METHODS AND MATERIALS: Between October 1980 and December 1990, 289 patients were treated with lumpectomy and/or axillary dissection, radical irradiation with or without adjuvant therapy. The radiation therapy technique consisted of external beam therapy followed by an Ir-192 implant or electron beam boost. Univariate and multivariate analysis were used to evaluate the correlation between multiple factors affecting the cosmetic outcome and the complications. RESULTS: Of 289 patients, cosmesis was evaluable in 285 and recorded as excellent to good in 226 (79%), fair in 35 (12%), and poor in 24 (9%). On multivariate analysis of these results, a high dose per fraction (p = .0018), and use of electrons as boost to the tumor bed (p = .001) were found to be significant. When patients boosted with electrons were excluded, a high boost dose (p = .0433) was also found significant. Fifteen (5%) patients developed severe late radiation sequelae and 14 (5%) moderate or severe arm edema. On multivariate analysis only dose per fraction of 2.5 Gy (p < .0001) and higher boost doses (p = .017) were significant. CONCLUSION: In 289 patients of early breast cancer treated with conservative surgery and radical irradiation, multivariate analysis suggests that higher dose per fraction with teletherapy and higher brachytherapy boost dose adversely affect cosmesis and contribute to the late complications. PMID- 8407403 TI - Poor survival of black patients in carcinoma of the endometrium. AB - PURPOSE: To compare the prognostic factors and survivals of black and white patients with endometrial carcinoma. METHODS AND MATERIALS: A retrospective study was undertaken of a total of 290 patients with endometrial carcinoma who were treated similarly at the Health Science Center at Brooklyn and Kings County Hospital Center from 1975 and 1990. One hundred and thirty-six of 290 (47.2%) were black and 135/290 (46.9%) were white. Well-known prognostic factors affecting endometrial carcinoma were studied in black and white group of patients. Their overall survival and comparison of survival in each prognostic group were also estimated using multi-variate analysis. RESULTS: Fifty-four percent of white patients had Stage I disease, compared to 45.9% in black patients. In Stage II, 51.6% were white and 48.4% were black, and in Stage III, 88.89% were black and 11.1% were white patients (p = 0.034). Fifty six percent Grade 1 patients were white and 44% were black. In Grade 2, 53.3% were white and 46.7% were black and in Grade 3 disease, 70.5% were black and 29.5% were white (p = 0.008). Up to the inner third of myometrial invasion had occurred in 60.6% of white patients and 39.4% in black patients. The middle third of the myometrium was invaded in 60.7% of white patients, and 39.3% of black patients. Thirty-seven percent of outer third of myometrial invasion was found in white patients and 63% in black patients (p = 0.038). Seventy-two percent of positive lymph nodes were found in black patients and 28.0% in white patients (p = 0.01). Sixty-one percent of patients with positive peritoneal cytology were black as compared to 38.7% in white patients (p = 0.017). The overall ten-year corrected survival for white and black patients was 72% and 40%, respectively (p = 0.0003). Survivals comparisons, when stratified by race and each prognostic group, showed statistically significant overall survival differences in favor of white patients. CONCLUSION: Black patients with endometrial carcinoma have poor survival. Low socio-economic status (SES) would not explain these findings. More research is required to determine the cause of poor survival in black patients with endometrial carcinoma. PMID- 8407404 TI - Intratumoral heterogeneity of malignant gliomas measured in vitro. AB - PURPOSE: To evaluate the extent of intratumoral heterogeneity of radiation sensitivity in malignant gliomas, by comparing the intrinsic radiation sensitivity of different glioma sublines derived from the same tumor. METHODS AND MATERIALS: The study was performed on five early established malignant gliomas (passage 3-10). Each specimen was quickly cut into three equal pieces (except for one specimen, where only two pieces were obtained). Each piece was processed independently, disintegrated into single cell suspension using a cocktail of enzymes. Survival curve assays, using colony formation as an end-point, were performed for each subline. Comparison between the intrinsic radiation sensitivity of sublines was calculated using the surviving fraction at 2 Gy and the mean inactivation dose as the measured parameters. The DNA content of the cell lines as well as their cell cycle analysis was determined using flow cytometry. RESULTS: The mean calculated surviving fraction at 2 Gy of all the sublines was 0.37 +/- 0.14, the mean mean inactivation dose was 1.98 +/- 0.63. The intertumoral coefficient of variation for the calculated surviving fraction at 2 Gy of all cell lines was 38%, while that for intratumoral heterogeneity was 25%. Three of the 5 tumors showed a statistically significant difference in the surviving fraction at 2 Gy and mean inactivation dose values of their sublines (p < 0.05). This difference in radiation sensitivity between sublines of the same tumor was not attributed to a difference either in the ploidy status or in the distribution of cells in the cell cycle. CONCLUSION: There is a significant intratumoral heterogeneity of radiation sensitivity in some malignant gliomas. This heterogeneity may limit the predictive power of surviving fraction at 2 Gy or mean inactivation dose, especially when their values are based upon a single measurement/single biopsy. In the meantime, this heterogeneity may be a factor in the discrepancy between unexpectedly sensitive tumor cell lines in vitro and their high clinical radiation resistance. PMID- 8407405 TI - The effect of fractionated doses of radiation on mouse spinal cord. AB - PURPOSE: To determine: (a) the dose-response relationship and latent time to paralysis following fractionated doses of radiation in mice, (b) the values of parameters for isoeffect curves, and (c) whether these parameters depend on the size of dose per fraction and the severity of injury. METHODS AND MATERIALS: The spinal cords (T9-L5) of 608 C3Hf/Sed/Kam mice were irradiated with fractionated doses of x-radiation. Three levels of neurological damage were used to grade the spinal cord response. Animals which did not develop paralysis were observed for at least 18 months after irradiation. The fractionated schedules consisted of either 2, 3, 4, 6, 10, or 20 fractions in addition to single doses. For the fractionated regimes the daily fraction size ranged from 2 Gy to 24 Gy, and for single doses the range was 12 Gy to 52 Gy. Both the latent time to paralysis and the incidence of paralysis were considered as endpoints. For analysis of the sparing associated with fractionation, the dose points were divided into two groups: a "low damage" group consisting of doses of near or less than the ED50 at 450 days and a "high damage" group consisting of doses much larger than the ED50 at 450 days in which there was 100% incidence of paralysis. RESULTS: The latent time depended on the radiation dose; for each fixed fraction number the latent period became progressively shorter with higher total doses. Differences in histology in fractionation sensitivity are observed between the two groups. The low damage data in each fractionation treatment are the important data in the analysis of long-term incidence of paralysis. On the other hand, the high damage data were emphasized for the analysis of latency. Three statistical methods (mixture model, Cox model, and Fe-plot) were used to fit the linear-quadratic dose response model and the "Nominal Standard Dose" (NSD) model. The values of the parameters of these two models depended on the effect evaluated; the latent interval from the high damage region being not very fractionation-dependent, whereas, the incidence of paralysis from the low damage fractionation regimens was strongly dependent on dose per fraction. Specifically, the alpha/beta ratios for latency were large (e.g., 17 to 57 Gy) when fractionation schemes in the high damage region were emphasized. If data from the fractionation schemes in the lower damage region with fraction size less than 15 Gy were emphasized, the alpha/beta ratios for incidence of paralysis were 3.3 (1.8, 6.0, 95% C.I.), 4.1 (2.8, 5.5), and 4.4 Gy derived by the mixture, Cox, and Fe-plot models, respectively. These "low damage" alpha/beta ratios were similar for all levels of injury from mild to complete paralysis, and are those which are more relevant to clinical radiotherapy. The coefficients for the "Nominal Standard Dose" formula in the present study were 0.33 +/- 0.01 (s.e.) (by the Strandqvist-type plot), 0.38 (the Cox model), or 0.40 (the mixture model) for level 2 injury at 450 days. CONCLUSION: The values of parameters in the isoeffect models were different when the data analyzed were derived from regimens using fractionated low or high damage doses. PMID- 8407406 TI - Cytotoxicity of restriction enzyme-induced DNA strand breaks in radiosensitive and radioresistant human tumor cell lines. AB - PURPOSE: To examine the role of sensitivity to specific types of DNA double strand breaks in human tumor cell response. METHODS AND MATERIALS: The X ray sensitive human squamous carcinoma cell line SCC-61 and the X ray-resistant line SQ-20B were exposed to the restriction enzymes HaeIII, HinfI, PvuII, BamHI by electroporation. Cytotoxicity of these restriction endonucleases was measured by a colony formation assay. RESULTS: Cell killing by each enzyme occurred in a concentration-dependent manner. The radiosensitive cell line was more sensitive to all four restriction enzymes than the radioresistant line, paralleling the response to ionizing radiation. However, the magnitude of the difference was smaller than for radiation. The 5-base sticky ended cutter HinfI and 6-base blunt ended cutter PvuII were much more effective in killing cells from both lines than BamHI, a 6-base sticky ended cutter, whereas the 4-base blunt ended cutter HaeIII was intermediate in its effectiveness. Thus, enzyme sensitivity could not be related to the type of cutter or the distance between cutting sites. PMID- 8407407 TI - Radiobiological modeling of combined targeted 131I therapy and total body irradiation for treatment of disseminated tumors of differing radiosensitivity. AB - PURPOSE: A model is presented for calculating combinations of targeted 131I and total body irradiation, followed by bone marrow rescue, in the treatment of tumors of different radiosensitivity. The model is used to evaluate the role of the total body irradiation component in the optimal combination regime as a function of the radiosensitivity of the tumor cells. METHODS AND MATERIALS: A microdosimetric model was used to calculate absorbed dose in small tumors and micrometastases when uniformly targeted by the radionuclide 131I. Cell kill was calculated from absorbed dose using an extended version of the linear quadratic model. The addition of varying total doses of total body irradiation, assuming 2 Gy fractions, was also calculated using the linear quadratic model. The net cell kill from combined modality (targeted 131I and total body irradiation) was computed for varying proportions of the two components, for a range of tumor sizes, restricting the total radiation dose to within tolerance for a full-course TBI regime (approximately 14 Gy total) in all cases. The calculations were repeated for a range of presumed tumor uptakes of the targeting agent and for a range of tumor radiosensitivities, typical of those reported for tumor cells of differing type in culture. Optimal regimes were identified as those predicted to yield a high probable tumor cure rate (evaluated using a Poisson statistical model) for all tumor sizes. RESULTS: The analysis supports earlier model studies which predicted that systemic combination treatment with targeted 131I and total body irradiation would be superior to either component used alone. The intrinsic tumor radiosensitivity is found to be a factor which influences the optimal combination of the 131I and external beam total body irradiation components. The total body irradiation component is greater in optimal regimes treating radio resistant than radiosensitive tumors. However, an obligatory total body irradiation component is also predicted for more radiosensitive tumors; the analysis suggests that the total body irradiation component should in no circumstances be less than 2 x 2 Gy, whilst practical arguments exist in favor of higher doses. CONCLUSION: Total body irradiation is an obligatory component for effective systemic treatment of disseminated malignant tumors to which 131I can be selectively targeted. Clinical studies applying this strategy to the treatment of neuroblastoma by 131I targeted by meta-iodo-benguanidine (mIBG), total body irradiation and bone marrow rescue are now in progress. PMID- 8407408 TI - Temperature differentials between treatment and pretreatment temperatures correlate with local control following radiotherapy and hyperthermia. AB - PURPOSE: To evaluate the influence of pretreatment tumor temperatures and the temperature differential between treatment and pretreatment temperatures on local tumor control in patients who underwent combined radiation therapy and hyperthermia. METHODS AND MATERIALS: Mapped intratumoral temperatures were measured immediately prior to and during hyperthermia in 138 hyperthermia fields among 59 patients with nodular (60 fields) or diffuse (78 fields) superficially located tumors. In the nodular subgroup there were 40 fields with adenocarcinomas (31 breast, two prostate, seven other primary sites), six melanomas, nine squamous cell carcinomas, and five other histologies. The fields with diffuse tumor involvement consisted of 77 adenocarcinomas (67 breast, 10 other) and one melanoma. The maximum, minimum, and average temperatures were determined for both the pretreatment (pTmax, pTmin, pTave) and treatment (Tmax, Tmin, Tave) distributions and the differences, Dm = Tmin-pTmax, and Da = Tmin-pTave, computed. These quantities were averaged over treatments to produce the corresponding mean quantities for each hyperthermia field. Univariate and multivariate analyses were performed to determine treatment and pretreatment parameters which best correlated with the duration of local control. RESULTS: Pretreatment tumor temperatures were significantly lower than the oral temperatures with mean pTmax, mean pTmin, and mean pTave of 36.2 degrees C, 34.2 degrees C, and 35.4 degrees C, respectively. For the adenocarcinomas with diffuse involvement within the hyperthermia field, the covariates best correlating with local control duration on univariate analysis were concurrent radiation dose (p = 0.0026), Dm (p = 0.009), pTmax (p = 0.012) and Da (p = 0.036). Lower pTmax and larger Dm and Da were predictive for longer local control. In multivariate analyses, all thermal parameters lost power, however, the best model included Dm which was significant at the p = 0.040 level. For the nodular subgroup, nonthermal parameters and dichotomized thermal parameters were of prognostic significance for local control. CONCLUSION: For fields diffusely involved with adenocarcinoma significant correlations with duration of local control have been demonstrated both for a) low pretreatment temperatures and b) large differentials between treatment and pretreatment intratumoral temperatures. These correlations were also found in a dichotomized description for fields with nodular tumors. The results support the concept that pretreatment hypothermic conditions can lead to an increase in thermal sensitization and may help explain the excellent clinical results noted in the treatment of superficial tumors with radiation and hyperthermia. Further exploitation of this approach by planned cooling of superficially-located recurrent tumors prior to hyperthermia treatment warrants investigation. PMID- 8407409 TI - Effect of set-up error on the dose across the junction of matching cranial-spinal fields in the treatment of medulloblastoma. AB - PURPOSE: The effect of systematic and stochastic setup error on the dose delivered to the gap region for the three field radiation treatment of medulloblastoma is studied. The consequences of such setup error is discussed. METHODS AND MATERIALS: The treatment of medulloblastoma is typically a 3 field technique, in which two lateral cranial fields are matched with a spine field. The x-ray dose delivered to the region between the matched fields depends upon the gap size. The choice of the gap width between the cranial and spinal fields is controversial. It is currently a compromise between minimizing the risk of dose hot spots to the spine, and the associated clinical complications, as well as the magnitude of cold spots (underdosing) across the gap, with the associated risk of disease recurrence. In this paper, we examine the effect of gap width with a moving junction, referred to as "field feathering", on the dose across the field junction for a 6MV photon beam. In addition, we have studied 129 portal films and 40 simulation films to assess the accuracy and precision of patient setup during treatment with a plan involving feathered fields. Selected landmarks observable on both portal and simulation films were identified and the variation in the distances to the field edges measured. The distribution of patient setup error was convoluted with the beam profiles for a 6MV linac. These convoluted field edges were used obtain dose profiles across the gap region as a function of gap separation. The consequences for therapy are discussed. In addition, analysis of patient setup error on an alternative treatment involving beam modifiers to broaden the beam penumbra is discussed. RESULTS: The magnitude of the spatial stochastic and systematic setup error was determined to be approximately three and two millimeters respectively. The dosimetric consequences of patient setup error lead to over and under dosing in the spinal gap region for the three field technique. The degree of under or over dose depends on the nature and magnitude of the patient setup error. CONCLUSIONS: The effect of patient setup error can lead to significant dosimetric errors in the dose to the gap region depending on the magnitude of the setup errors. The effective over and under dose can be compensated by the use beams modifiers such as a beam spoiler or vibrating jaws. PMID- 8407410 TI - A theoretical analysis of hemodynamic and biomechanical alterations in intracranial AVMs after radiosurgery. AB - PURPOSE: Stereotactic radiosurgery is being increasingly used to treat intracranial arteriovenous malformations (AVMs). However, successful radiosurgery may involve latent periods of 1-2 years prior to AVM obliteration. This latent period include states of altered flow patterns that may or may not influence hemorrhage probabilities. The probability of hemorrhage is likely to be related to the degree of biomechanical stress across the AVM shunt walls. This paper describes a theoretical analysis of the altered hemodynamics and biomechanical stresses within AVM shunts post-radiosurgery. METHODS AND MATERIALS: The mathematical model is comprised of linked flow compartments that represent the AVM and adjacent normal vasculature. As obliteration of the irradiated shunts occur, changes in flow rates and pressure gradients are calculated based on first order fluid dynamics. Stress on the AVM shunt walls is calculated based on tangential forces due to intramural pressure. Two basic models are presented: a distribution of shunts with fixed thin walls subject to step-function obliteration (Model I), and a distribution of shunts subject to luminal obliteration from slowly thickening walls (Model II). Variations on these models are analyzed, including sequential, selective and random shunt obliteration, and uniform or Poisson distributions of shunt radii. RESULTS: Model I reveals that the range of pressure alterations in the radiosurgically-treated AVM include the possibility of transient increases in the total biomechanical stress within the shunt walls prior to obliteration. Model II demonstrates that uniform luminal narrowing via thickened walls should lead to reduced transmural stresses. The precise temporal pattern of AVM flow decrease and biomechanical stress reduction depends on the selection of shunts that are obliterated. CONCLUSION: (a) The hemodynamic and biomechanical changes appear to be relatively independent of the shunt distribution but highly dependent on the temporal pattern of the obliterative process, (b) uniformly thickened shunt walls should uniformly decrease biomechanical stresses in the latent period prior to complete obliteration, but if uniform obliteration is not achieved, (c) transient alterations in pressure versus stress relationships may lead to temporarily increased biomechanical stress prior to complete obliteration, and (d) reduction in stress may not reach significant levels until the AVM is almost completely obliterated. PMID- 8407411 TI - Dose intensification in curative head and neck cancer radiotherapy--linear quadratic analysis and preliminary assessment of clinical results. AB - PURPOSE: The feasibility of reducing overall treatment time by 2 weeks in the curative radiotherapeutic management of head and neck cancer patients is reported in a pilot trial of Hyperfractionated, Accelerated Radiotherapy with Dose Escalation (HARDE). This regimen prescribes 76 Gy in 5 weeks to definitive head and neck cancer patients, and 65 Gy in 5 weeks to high-risk postoperative patients. The linear quadratic model is used to compare predicted tumor cell kill with HARDE versus that expected with conventional fractionation (CF). MATERIALS AND METHODS: Between January 1991 and March 1992, 40 head and neck cancer patients were treated with HARDE at the University of Wisconsin Comprehensive Cancer Center. Case-matched controls treated with CF were identified from patients treated at the same institution between 1980-1990, based on tumor site, stage, and extent of prior surgery. Individual patient treatment data (total dose, fraction size, overall time) rather than idealized schedule data from each group were analyzed using the linear quadratic model. RESULTS: Seventy-nine case matched controls were identified for comparison with HARDE patients. The predicted increase in log cell kill for HARDE patients over case-matched controls was 1.5 and 1.3 logs, respectively, in the definitive and postoperative settings. This difference in log cell kill projects an improvement in locoregional tumor control for HARDE patients of between 10-25%. HARDE patients experience very brisk acute mucosal reactions and moderately prolonged mucosal healing, however, 91% have completed therapy without a treatment break. CONCLUSION: A 2-week reduction in overall treatment time for curative head and neck cancer patients is feasible while maintaining doses > 70 Gy. Based on radiobiologic predictions, such treatment intensification may significantly improve rates of locoregional tumor control. However, intensified acute mucosal reactions accompany such accelerated therapy. PMID- 8407412 TI - Synchronous radiotherapy and chemotherapy in the treatment of nasopharyngeal carcinoma. AB - PURPOSE: Because of the high rates of local tumor control obtained by combining moderate doses of external beam radiotherapy and synchronous 5 fluorouracil/mitomycin C chemotherapy in the treatment of squamous and basiloid cancers of the anal canal, we chose to investigate this regimen for nasopharyngeal cancer which shows significant local and distant failure rates after treatment with radiotherapy alone. METHODS AND MATERIALS: Between 1983 and 1990, 43 patients with previously untreated squamous cell and undifferentiated nasopharyngeal cancer, without evidence of distant metastases at diagnosis were treated with radical radiotherapy and concurrent chemotherapy using mitomycin C (10 mg/m2 i.v. day 1 of radiotherapy) and 5-fluorouracil (1000 mg/m2 continuous i.v. infusion days 1-4 of radiotherapy and repeated at least 28 days later). Ninety-one percent of cases had Stage IV tumors and 93% had clinically involved regional lymph nodes. RESULTS: Actuarial rates of survival, local control, regional nodal control and distant metastases at 5 years were 37%, 71%, 94%, and 53%. Grade 3 or 4 skin and mucosal reactions occurred in 30% and 34% of patients, respectively. Only one patient developed greater than Grade 2 myelosuppression and he died of overwhelming sepsis. A second patient died of malnutrition 4 months after treatment giving a 5% incidence of treatment-related mortality. Nine percent of patients developed significant late complications of treatment. CONCLUSION: Despite the morbidity observed, the treatment outcome is not obviously superior to that reported for radiotherapy as a single modality of treatment. PMID- 8407413 TI - Carcinoma in situ of the glottic larynx: the role of radiotherapy. AB - PURPOSE: To examine the University of Florida experience with radiotherapy of carcinoma in situ of the true vocal cord and compare our results with those reported in other series using various treatment modalities. METHODS AND MATERIALS: Nineteen patients with carcinoma in situ of the true vocal cord were treated with curative intent with radiation therapy at the University of Florida between October 1964 and September 1990. All patients had a minimum 2-year follow up. Before the radiotherapy, four patients had undergone a biopsy only, 11 patients had undergone one stripping procedure, and four patients had undergone 2 to 5 strippings. Radiation doses were between 5625 cGy and 6300 cGY (median, 5625 cGy). RESULTS: The 5-year rates of local control, ultimate local control, and local control with voice preservation were 93%, 100%, and 93%, respectively. A literature review revealed that local control rates after primary treatment with radiotherapy, laser resection, and vocal cord stripping were 84%, 68%, and 66%, respectively. CONCLUSION: Primary treatment with radiotherapy should be strongly considered for patients with carcinoma in situ of the true-vocal cord who have a recurrence after vocal cord stripping or who cannot have close follow-up after treatment. PMID- 8407414 TI - Radiotherapy of cutaneous B cell lymphomas: our experience in 31 cases. AB - PURPOSE: Since cutaneous B cell lymphomas belong mostly to low or intermediate histologic grade of malignancy and have low tendency to spreading, a local treatment such as radiotherapy appears to be a suitable choice in the management of the disease. The authors have reviewed their cases to verify this statement. METHODS AND MATERIALS: 31 patients affected by cutaneous B cell lymphomas classified as IE stage according to Ann Arbor received radiotherapy with orthovoltage techniques with total doses per field ranging from 10 to 40 Gy (median dose 30 Gy). All the patients had a minimum follow-up of 2 years. RESULTS: All the target skin lesions underwent complete remission. In 10 patients (32.2%) the clinical remission is still lasting. In 21 cases (67.8%) a disease relapse was observed: only at skin in other sites than those previously treated in 17 (81%), at skin and lymph nodes in two cases (9.5%), at skin, bone, and lymph node in one case (4.7%), at skin and bowel in one case (4.7%). The extracutaneous involvement occurred in cases with lesions of intermediate grade malignancy. After a new course of radiotherapy for skin lesions only, and chemotherapy, surgery or megavoltage radiotherapy for the other involvements, on the whole 21 patients (67.8%) got a complete remission. CONCLUSION: On the basis of their results and of a review of the literature, the authors propose radiotherapy as the choice treatment of primary cutaneous B cell lymphomas. PMID- 8407415 TI - Total skin electron therapy: a technique which can be implemented on a conventional electron linear accelerator. AB - PURPOSE: A technique for treating whole-body skin with an electron linear accelerator with nominal energies in the range 4-8 MeV is presented. Stationary fields at an extended source-skin distance are used with the patient treated in a reclined position. METHODS AND MATERIALS: The relative beam data, absolute dosimetry measurements and the patient setup parameters are presented. The calculations required to correct for patient size are discussed. RESULTS: The technique described uses a six field circumferential cycle, with longitudinally matched fields along the length of the patient. Treatment times are reasonable using the standard dose rate of the machine. The uniformity of the skin dose measured on three patients was found to be comparable to that of other total skin treatment techniques. CONCLUSION: A technique for treating conditions like mycosis fungoides is presented requiring relatively simple supporting dosimetry. No modifications to the unit are required and no sophisticated treatment apparatus is necessary, which makes the technique attractive to smaller Institutions, especially in developing countries, where technical support may be limited. PMID- 8407416 TI - Potential human error in setting stereotactic coordinates for radiosurgery: implications for quality assurance. AB - PURPOSE: The error frequency in setting stereotactic coordinates for gamma knife radiosurgery was investigated to determine what quality assurance safeguards are necessary. METHODS AND MATERIALS: A prospective study of 200 consecutive isocenter settings for gamma knife radiosurgery was analyzed to identify the frequency of spontaneous errors in setting and checking stereotactic coordinates (corrected prior to treatment). An additional 25 coordinate errors were introduced at random among the next 200 consecutive isocenter settings to provide additional data on identification of errors. RESULTS: Stereotactic coordinates required resetting in 12% (24/200) of the isocenters treated due to errors of 0.25-0.50 mm (8%) and 1-20 mm (4%). This comprised 2.2% (26/1200) of the individual coordinate settings. The frequency of these errors was significantly related to the specific directional coordinate set (p = 0.0004) and experience (p = 0.016). Errors were identified by 83.5% (91/109) of the observers checking the settings (60.0% of 0.25 mm errors, 94.6% of errors > or = 0.5 mm, p = 0.0000). Verification of stereotactic coordinates by two observers reduces the probability of an undetected error > or = 0.25 mm to 1/1,392 and to 1/154,712 for errors > or = 1 mm. CONCLUSION: Errors in setting stereotactic coordinates are common (12% prior to checking) but are corrected with a high degree of confidence by a quality assurance policy requiring coordinate verification by a minimum of two observers. PMID- 8407417 TI - Doses to radiation sensitive organs and structures located outside the radiotherapeutic target volume for four treatment situations. AB - This study documents dosage to radiation sensitive organs/structures located outside the radiotherapeutic target volume for four treatment situations: (a) head and neck, (b) brain (pituitary and temporal lobe), (c) breast and (d) pelvis. Clinically relevant treatment fields were simulated on a tissue equivalent anthropomorphic phantom and subsequently irradiated with Cobalt-60 gamma rays, 6- and 18-MV x-ray beams. Thermoluminescent dosimeters and diodes were used to measure absorbed dose. The head and neck treatment resulted in significant doses of radiation to the lens and thyroid gland. The total treatment lens dose (300-400 cGy) could be cataractogenic while measured thyroid doses (1000-8000 cGy) have the potential of causing chemical hypothyroidism, thyroid neoplasms, Graves' disease and hyperparathyroidism. Total treatment retinal (400 700cGy) and pituitary (460-1000 cGy) doses are below that considered capable of producing chronic disease. The pituitary treatment studied consisted of various size parallel opposed lateral and vertex fields (4 x 4 through 8 x 8 cm). The lens dose (40-200 cGy) with all field sizes is below those of clinical concern. Parotid doses (130-1200 cGy) and thyroid doses (350-600 cGy) are in a range where temporary xerostomia (parotid) and thyroid neoplasia development are a reasonable possibility. The retinal dose (4000 cGy) from the largest field size (8 x 8 cm2) is in the range where retinopathy has been reported. The left temporal lobe treatment also used parallel opposed lateral and vertex fields (7 x 7 and 10 x 10 cm). Doses to the pituitary gland (5200-6200 cGy), both parotids (200-6900 cGy), left lens (200-300 cGy) and left retina (1700-4500 cGy) are capable of causing significant future clinical problems. Right-sided structures received insignificant doses. Secondary malignancies could result from measured total treatment thyroid doses (670-980 cGy). Analysis of three breast/chest wall and regional nodal irradiation techniques demonstrated a 25-50% decrease in secondary lung dose with use of independent collimation compared to use of custom alloy blocking material. However, it is unlikely that a reduction in secondary dose of this magnitude would reduce the risk of treatment sequellae. In four-field "box" pelvic irradiation, secondary testes dose may result in temporary (clamshell shield) or permanent azoospermia, but is unlikely to impair androgen production. PMID- 8407418 TI - The influence of air cavities on interface doses for photon beams. AB - PURPOSE: As the quantification of dose in homogeneous media is now better understood, it is necessary to further quantify effects from heterogeneous media. The most extreme case is related to air cavities. Although dose corrections at large distances beyond a cavity are accountable by attenuation differences, perturbations at air-tissue interfaces are complex to measure or calculate. These measurements helps understand the physical processes that govern these perturbations. METHODS AND MATERIALS: A thin window parallel-plate chamber and a special diode were used for measurements with various air cavity geometries (layer, channel, cubic cavity, triangle) in x-ray beams of 4 and 15 MV. RESULTS: Underdosing effects occur at both the distal and proximal air cavity interfaces. The magnitude depends on geometry, energy, and field sizes. As the cavity thickness increases, the central axis dose at the distal interface decreases. Increasing field size remedied the underdosing, as did the introduction of lateral walls. Following a 2.0 cm wide air channel for a 4 MV, 4 x 4 cm2 field there was an 11% underdose at the distal interface, while a 2.0 cm cubic cavity yielded only a 3% loss. Measurements at the proximal interface showed losses of 5% to 8%. For a 4 MV parallel opposed beam irradiation the losses at the interfaces were 10% for a channel cavity (in comparison with the homogeneous case) and 1% for a cube. The losses were slightly larger for the 15 MV beam. Underdosage at the lateral interface was 4% and 8% for the 4 MV and 15 MV beams, respectively. CONCLUSION: Although reports suggest better clinical results using lower photon energies with the presence of air cavities, there is no reliable dose calculation algorithm to predict interface doses accurately. The measurements reported here can be used to guide the development of new calculation models under nonequilibrium conditions. This situation is of clinical concern when lesions such as larynx carcinoma beyond air cavities are irradiated. PMID- 8407419 TI - Entrance and exit dose regions for a Clinac-2100C. AB - PURPOSE: While significant advances have taken place in confirming dose in homogeneous media and accounting for changes due to distant heterogeneities, interface dosimetry is still a dose assessment problem. The entrance and exit surfaces of the patient are a prime example where dose assessment is questionable, but important, in many clinical situations. METHODS AND MATERIALS: Data were taken to examine the effect of such parameters as field size, source-to skin distance, blocking tray, compensation materials (lead, aluminum and brass) and various patient support materials (Mylar, graphite, thermal plastic, and foam) on the surface doses from a dual energy (6 MV, 18 MV) linear accelerator. Measurements were conducted with a thin window parallel-plate chamber. RESULTS: Relative surface dose was found to have a proportional dependence on field size for both energies with surface doses ranging from 6% to 53%. A surface depth dose of 22.6% for a 15 cm field was measured for the low energy beam while a surface dose of 22.3% was observed for the 18 MV beam. The surface dose increased significantly for short source to skin distances and with the presence of a blocking tray. Compensating filter materials had little influence on surface dose. However, patient support devices such as Alpha Cradle and the graphite of the table can increase the surface dose to as much as 92%. CONCLUSION: We found there was a loss in dose at the exit surface (in comparison with percent depth dose tables, which assume infinite depth) on the order of 15% and 11% for the 6 MV and 18 MV beams, respectively. However, this loss is quickly compensated for with the introduction of most inherent backscattering media, [which in fact can increase the dose] for example, graphite in the patient support assembly tabletop. PMID- 8407420 TI - The effect of setup uncertainties on the treatment of nasopharynx cancer. AB - PURPOSE: As radiation treatment techniques become more complicated, the need to understand the effect of uncertainties on dose distributions increases. This study investigates the effect of positional uncertainities for patients with nasopharynx carcinoma treated with a multiple field conformal boost technique. Three dimensional setup errors were measured for six patients and the effect on patient dose was evaluated using dose volume histograms. METHODS AND MATERIALS: A method is presented for determining 3-dimensional translational and rotational setup errors by identifying anatomical landmarks on two treatment field images and their corresponding simulation images. Measurements were made on a daily basis for six patients undergoing conformal treatment. RESULTS: The average magnitude of the translational errors was between 1.5 and 3 mm while the average distance between simulation and treatment isocenters was 5 mm. Both systematic and random setup errors were observed. Dose volume histograms incorporating these uncertainties for standard parallel opposed and conformal techniques were generated for patients experiencing random and systematic setup errors. CONCLUSION: The data imply that positional uncertainties effect the daily dose distributions for target and critical structures differently and that the effect may be treatment technique dependent. These results demonstrate the need to measure setup uncertainties for all sites and to develop techniques for incorporating dose uncertainties in treatment plans. PMID- 8407421 TI - Verification of lung attenuator positioning before total body irradiation using an electronic portal imaging device. AB - PURPOSE: We report the first clinical experience with an electronic portal imaging device for lung attenuator positioning before delivery of total body irradiation. We demonstrate a technique for lung attenuator placement which reduces the dose to the patient during setup, reduces the patient setup time, and increases the accuracy of lung attenuator positioning. METHODS AND MATERIALS: Patients are treated with total body irradiation using a dedicated dual source irradiation facility prior to receiving bone marrow transplantation. The dose rate to the patient's midline is limited to 0.10 Gy/min, and partial transmission lung blocks are used to minimize radiation induced pneumonitis while delivering adequate dose to the regions under the blocks. Lung blocks are placed on the patient's back and chest wall, and portal images are used to verify proper block placement before the remaining treatment dose is delivered. RESULTS: We report the use of a liquid ionization chamber matrix electronic portal imaging device for imaging total body irradiation patient setups. CONCLUSION: The dose to the patient using the EPID for portal imaging is a factor of 7.5 lower than that needed for film. Image quality is superior to that of film due to digital processing. Since less time and dose are needed for imaging, it is demonstrated that better and more efficient final placement of the lung blocks can be achieved. PMID- 8407422 TI - Positron emission tomography with fluorodeoxyglucose to evaluate tumor response and control after radiation therapy. AB - PURPOSE: Following radiation therapy, evaluation of viable tumor can often be difficult with anatomic imaging criteria (tumor size alone). In this study, the utility of biochemical imaging with the glucose analog 2-[F-18]fluoro-2-deoxy-D glucose and positron emission tomography was investigated in patients treated with radiation therapy. METHODS AND MATERIALS: Between 1990 and 1992, 19 patients were studied, including 15 patients with head and neck cancer, (4 oropharynx, 4 sinus, 3 larynx, 2 hypopharynx, 2 oral cavity [one patient], 1 nasopharynx), and 4 patients with breast cancer. Post-radiation positron emission tomography with 2 [F-18]fluoro-2-deoxy-D-glucose studies were done in all patients, with 9 head and neck patients receiving pre-radiation positron emission tomography with 2-[F 18]fluoro-2-deoxy-D-glucose scans as well. Results were correlated with other imaging techniques and pathology. RESULTS: Positron emission tomography with 2-[F 18]fluoro-2-deoxy-D-glucose detected head and neck primary tumors and lymph node metastases in all nine pre-radiation scans, while magnetic resonance imaging failed to detect two primary tumors. Serial positron emission tomography with 2 [F-18]fluoro-2-deoxy-D-glucose showed a significant decrease in tumor activity after radiation therapy, compared to pre-radiation levels, (p < 0.05), except for two patients with increased uptake at the primary site. Biopsies of these two patients showed persistent/recurrent disease after radiation therapy, which was not detected by magnetic resonance imaging. Six additional head and neck patients, with suspicious examination and inconclusive magnetic resonance imaging, were imaged with positron emission tomography after radiation therapy only. Five patients had increased positron emission tomography activity, with corresponding biopsies positive in four patients, and negative in one patient with clinically worsening symptoms. The remaining sixth patient had minimal and stable positron emission tomography uptake, and is improving clinically. Four patients had mammogram findings suspicious for recurrence after conservation treatment for breast cancer. Positron emission tomography with 2-[F-18]fluoro-2 deoxy-D-glucose showed no focal activity in the breast in two patients, and increased activity in the area suspicious for recurrence in the other two patients. Biopsies correlated with positron emission tomography results. CONCLUSION: Changes and presence of positron emission tomography with 2-[F 18]fluoro-2-deoxy-D-glucose activity correlated with pathologic findings in head and neck and breast cancer patients in this series. In patients with elevated or rising positron emission tomography activity after radiation therapy, persistent or recurrent disease was found in 89% of patients, (8/9). Magnetic resonance imaging did not detect the head and neck recurrences, and mammography was suspicious in patients with both benign and malignant breast changes after radiation therapy. In addition, our data indicate that in head and neck patients with pre-radiation positron emission tomography scans, a significant decrease in activity should occur after radiation therapy, if local control is to be expected. PMID- 8407423 TI - Cisplatin and radiotherapy in the management of locally advanced head and neck cancer. AB - PURPOSE: This manuscript traces the results of combined modality head and neck cancer trials, with reference to survival and rates of local-regional control and distant metastases, that have led to current Phase III trials testing concomitant cisplatin and radiotherapy. METHODS AND MATERIALS: The toxicity, local control rates, and survival results of pilot trials and small randomized trials using concomitant cisplatin based chemotherapy and radiotherapy are reviewed. RESULTS: Enhanced mucosal toxicity and myelosuppression occur which affect patient compliance and tolerance of planned chemotherapy, but do not appear to affect delivery of standard fractionation radiotherapy. The encouraging results of single agent cisplatin and radiotherapy trials and those using cisplatin-5-FU and split course radiotherapy have led to the activation of three randomized Head and Neck Intergroup trials. These trials are for Stage III & IV carcinoma of the nasopharynx, Stage III & IV resectable carcinoma of the larynx (organ preservation) and Stage III & IV unresectable carcinoma of the oral cavity, oropharynx, hypopharynx, and larynx. CONCLUSION: The three Intergroup study designs should allow for a definitive evaluation of concomitant treatment. PMID- 8407424 TI - Craniopharyngioma revisited. PMID- 8407425 TI - Cavernous sinus cranial neuropathies: is there a dose-response relationship following radiosurgery? PMID- 8407426 TI - The radiobiological basis of concomitant cisplatin and radiotherapy. PMID- 8407427 TI - Regarding Million's editorial on the "Relative clinical influence of tumor dose versus dose per fraction ... following larynx radiotherapy". PMID- 8407428 TI - Response to editorial by Dr. Earle. PMID- 8407429 TI - Global Field Power: a 'time-honoured' index for EEG/EP map analysis. PMID- 8407430 TI - Respiratory influences on the cardiac defense response. AB - An investigation to examine the relationships between breathing activity and the cardiac defense response (CDR) to intense auditory stimulation is reported. 42 subjects (20 men and 22 women) underwent a physiological reaction test consisting of three trials of a distorted 400 Hz noise of 100 dB, 0.5-s duration and instantaneous risetime presented either during inspiration or expiration. The respiratory response was characterized by a specific increase in breathing amplitude both in the respiratory cycle in which the stimulus was presented and in the 80 s following stimulus onset. Significant habituation effects were observed. Manipulation of the respiratory phase did not produce any major effect on the respiratory or cardiac response. The only significant finding was observed in the inspiratory period of the cycle in which the stimulus was presented, which was longer when the stimulus was presented in expiration and shorter when it was presented in inspiration. The evocation of the cardiac defense response was dependent on the observed increase in breathing amplitude. Gender differences were also observed in the respiratory response, differences which were further increased when the CDR was elicited. These results are discussed in the context of centrally versus peripherally-mediated mechanisms and startle versus defense reflexes. PMID- 8407431 TI - The effect of biologically-active light on the noo- and thymopsyche and on psychophysiological variables in healthy volunteers. AB - The study was carried out to show how bright light affects intellectual mnestic performance alertness, mood and psychophysiological variables in order to understand the mechanism of bright light which is still unclear. 15 healthy normal volunteers (8 males, 7 females) aged 22-34 years (mean 27) participated in the study. Exposure to bright light (2500 lux) or dim light (500 lux) was intermittent for 4 h between 9 a.m. and 5 p.m. Psychometric investigations for assessment of noopsychic variables, such as attention and concentration on the one hand, and thymopsychic variables, such as drive, on the other hand, as well as flexibility and psychophysiological measurements were carried out before, as well as 1, 2, 4, 6 and 8 h after baseline. As compared to the dim-light condition, subjects who were exposed to biologically-active light showed an improvement of noopsyche in the 6th and 8th hour. Also, the subjective thymopsyche revealed an improvement lasting for the whole investigation period. Psychophysiological measurements reflected an improvement of central and autonomous activation, which was parallel to the improvement of cognition and of well-being. However, biologically-active light did not affect flexibility. PMID- 8407432 TI - Habituation of phasic and tonic components of the orienting reflex. AB - This study examined phasic and tonic measures of electrodermal activity in a simple habituation paradigm with innocuous visual stimuli. Prestimulus skin conductance levels were taken as measures of the arousal level existing at each stimulus presentation, and the subsequent electrodermal responses were taken as indices of the phasic orienting reflex (OR) elicited by each stimulus. The arousal level was considered as a tonic OR measure. Both the tonic and phasic ORs showed effects of trials. Arousal levels also showed an initial sensitization, as suggested by dual-process theory. After within-subject effects of arousal changes over trials were removed by simple regression analyses, novelty still remained as the major determinant of phasic OR magnitude. This finding is discussed in relation to the OR mechanism. PMID- 8407433 TI - Electromyographic activation and proportion of fast versus slow twitch muscle fibers: a genetic disposition for psychogenic muscle tension? AB - 17 right-handed males volunteered for an experiment that compared task-related patterns of electromyographic (EMG) activation with data from muscle biopsy on proportion of slow-twitch ((ST) aerobic) to fast-twitch ((FT) anaerobic) muscle fibers. The biopsy was taken from the right-leg gastrocnemius muscle after EMG measurement from that area of the leg muscle. EMG was also recorded from the left forearm flexor carpi radialis area. Recordings were obtained from pre- and post task resting periods and during 150 s of video-task performance when the right hand operated a joy-stick. The results showed a highly significant tonic EMG activation in the leg muscle of subjects with predominance of ST fibers, and this relationship generalized to the EMG from the 'passive' forearm. The proportion of ST to FT fibers is genetically defined and not altered by exercise. Therefore, our results lend support to a genetic differentiation between individuals with high vs. low probability of unintended build-up of muscle tension during perceptual-motor task performance. PMID- 8407434 TI - Olfactory event-related potentials in psychosis-prone subjects. AB - Previous studies in schizophrenic patients have suggested that there are changes in olfactory sensitivity. In order to externally validate a psychometrical assessment of the psychosis-risk indicated by schizotypic factors, this study was carried out to determine whether changes in olfactory perception could be determined even for persons merely at risk of developing schizophrenia. These 'psychosis-prone' subjects consistently scored high in either the scale for 'physical anhedonia' (PA) or the scale for 'perceptual aberration' (PAB). Thus, three groups were investigated (control, n = 11; PA, n = 12; PAB, n = 12). Each subject participated in one testing session where the two odorants, vanillin (pleasant) and hydrogen sulphide (unpleasant), were applied by means of a specially designed delivery apparatus. Subjects rated both the intensity and the hedonic quality of the stimuli. In addition, olfactory event-related potentials (OERP) were recorded after dichotomous stimulation. In general, there were only few significant differences between the three groups investigated. Contrary to expectations, ratings for pleasantness of vanillin were highest in PA subjects compared to PAB subjects and controls (p < 0.05). Correspondingly, OERP amplitudes in response to vanillin were largest within the PA group (p < 0.05). For hydrogen sulphide, PAB subjects showed the smallest OERP amplitudes (p < 0.05). In addition, it was observed that female subjects had significantly larger OERP amplitudes when compared to male subjects (p < 0.05), which possibly indicates gender differences in olfactory sensitivity. PMID- 8407435 TI - EEG changes related to one-dimensional hand-tracking. AB - Central and occipital EEGs were recorded during one-dimensional eye-hand tracking in humans. A negative slow potential shift followed the commitment of tracking errors at electrode O1 and their correction both at O1 and C3. Error commitments were also followed by a relative decrease in alpha and beta-band power at O1, whereas their corrections followed a decrease in beta-band power at O2 and C3. PMID- 8407436 TI - Auditory-evoked responses influenced by presence or absence of EEG alpha activity and actual cognitive state. AB - The presence or absence of alpha activity in occipital EEG was detected automatically. Auditory-evoked responses (AERs) related to these two brain states and at the same time to different perceptual-cognitive situations (passive listening, discrimination between two tones and pressing the button accordingly similar discrimination without motor reaction) were analyzed and compared statistically. The main result was that during the alpha state (and not during the non-alpha state, when EEG was desynchronized), the amplitudes of N1, P1 waves and peak-to-peak amplitudes N1-P2 were higher during passive listening to stimuli in comparison with the active perceptual-cognitive states mentioned. No differences in latencies of AERs were found. PMID- 8407437 TI - Defensiveness, anxiety and the amplitude/intensity function of auditory-evoked potentials. AB - This study measured relationships between defensiveness, anxiety, and auditory evoked potentials to tones of varied intensity. Subjects were designated as defensive if they scored > or = 7 on the L-scale of the Eysenck Personality Questionnaire, and high-anxious if they scored > or = 11 on the N-scale. Four groups resulted: 'high anxious', 'defensive high anxious', 'repressors' (i.e., defensive low anxious) and 'low anxious'. Evoked potentials were recorded from FZ, CZ, PZ, C3, C4, T3 and T4, referenced to linked ears in response to 74, 84, 94 and 104 dB SPL tones. High-defensive subjects showed lower P2 amplitudes to the 94 and 104 dB tones and lower amplitude/intensity slopes at FZ, CZ, C3 and C4. High-anxious subjects showed lower P2 amplitudes to all four stimulus intensities at FZ, CZ and PZ. The results are consistent with the hypothesis that defensiveness is associated with desensitization to intense or painful stimulation. PMID- 8407438 TI - Eye movements, verbal material recall and negative symptoms in chronic schizophrenia. AB - Eye movements in response to verbal stimuli (Japanese cursive syllabary from the Standard Language test of Aphasia) were examined in 20 chronic schizophrenics diagnosed by DSM III and 15 normal controls. In chronic schizophrenics, the number of correct recalls in the test were lower than those in the normal controls, both the deviation indices and maximum deviation indices (the degree of fixation-time deviation) of chronic schizophrenics were higher than those of controls. Thus in chronic schizophrenics the fixation time for each character had greater deviation than in controls, and fixation on a single character was longer than in controls. In chronic schizophrenics there were significant negative correlations between syllables correctly recalled and both the deviation indices and the maximum deviation indices, while there were no significant correlations in controls. There were significant positive correlations between the four fixation time parameters; average fixation time on one character, deviation indices, maximum and minimum deviation indices and the severity of schizophrenia, especially that of negative symptoms. The results showed that chronic schizophrenics had distinct fixation time differences when presented with verbal stimuli, suggesting a mnemonic strategy deficit. The degree of fixation time deviation depended on the severity of negative symptoms. PMID- 8407439 TI - Heritability estimations of osteochondrosis in the tibiotarsal joint and of bony fragments in the palmar/plantar portion of the metacarpo- and metatarsophalangeal joints of horses. AB - Radiography of the tibiotarsal and metacarpo- and metatarsophalangeal joints was performed on 753 Standardbred trotters (6 to 21 months old) born in 1988. The surveyed population was drawn at random from all parts of Norway and represented about 60% of Standardbred trotters born the same year. Osteochondrosis in the tibiotarsal joint was diagnosed in 108 (14.3%) horses, and the prevalence of disease in progeny groups > 10 ranged from 0 to 69%. Bony fragments in the palmar/plantar portion of the metacarpo- and metatarsophalangeal joints were diagnosed in 89 (11.8%) horses, and the prevalence of disease in progeny groups > 10 ranged from 0 to 41%. Heritability analysis was restricted to 644 horses, comprising groups of 5 or more progeny, from 39 stallions. The heritabilities of osteochondrosis in the tibiotarsal joint and of bony fragments in the palmar/plantar portion of the metacarpo- and metatarsophalangeal joints were estimated to be 0.52 and 0.21, respectively, using a nonlinear model. PMID- 8407440 TI - Hemodialysis for treatment of oxytetracycline-induced acute renal failure in a neonatal foal. AB - Acute renal failure in a 4-day-old foal secondary to oxytetracycline toxicosis was treated by hemodialysis. Oxytetracycline had been administrered as treatment for forelimb flexor tendon contracture. Conservtive treatment with fluids, furosemide, and dopamine partially alleviated serum electrolyte concentration imbalances, but was ineffective in promoting diuresis or decreasing azotemia. Three hemodialysis treatments over 4 days were administered, after which the clinical appearance of the foal improved, and biochemical and electrolyte values returned to within reference ranges. The nephrotoxic potential of oxytetracycline should be considered prior to its administration. Hemodialysis may be a treatment option in management of acute renal failure in foals that are nonresponsive to conservative medical treatment. PMID- 8407441 TI - Surgical treatment of cervical stenotic myelopathy in horses: 73 cases (1983 1992). AB - Seventy-three horses with cervical stenotic myelopathy underwent cervical vertebral interbody fusion (n = 63) or dorsal laminectomy (n = 10). Neurologic function improved in 77% of horses, and 46% of horses achieved athletic function (racing, race training, or pleasure riding) after cervical vertebral interbody fusion for static and dynamic spinal cord compressive lesions. Neurologic status improved in 4 of 10 horses after dorsal decompression for static compressive spinal cord lesions. The duration of clinical signs prior to surgical intervention was shorter for horses that achieved athletic function or improved by at least 2 neurologic grades than for horses that did not improve in neurologic status or improved 1 neurologic grade after cervical vertebral interbody fusion. The number of cervical spinal cord compressive lesions and age of horses did not affect the long-term surgical outcome of cervical vertebral interbody fusion. Seroma formation, implant failure, right laryngeal hemiplegia, and colitis were nonfatal complications associated with cervical vertebral interbody fusion. Dorsal laminectomy and cervical vertebral interbody fusion of static compressive lesions of the caudal cervical vertebral column were associated with fatal postoperative complications, including vertebral body fracture, spinal cord edema, and implant failure. PMID- 8407442 TI - Effects of administration of recombinant bovine somatotropin on the responses of lactating and nonlactating cows to heat stress. AB - An experiment was performed to determine whether bovine somatotropin (BST) increases the severity of responses to heat stress and whether this effect depends on an increase in milk yield. Lactating and nonlactating cows received daily SC injections of BST (25 mg) or sodium phosphate solution for 12 days. Rectal temperatures and respiration patterns were not affected by BST when cows were maintained in a thermoneutral environment on day 9 of treatment. On day 12, cows were heat stressed by exposure to direct solar radiation. Rectal temperature, respiration rate, and the frequency of open-mouth panting increased as a result of heat stress. For lactating and nonlactating cows, those treated with BST had higher rectal temperatures and rate of open-mouth panting than cows treated with sodium phosphate solution. Administration of BST can increase the severity of responses of cows to heat stress without changing milk yield. PMID- 8407443 TI - Progress after one year of a pseudorabies eradication program for large swine herds. AB - Six large farrow-to-finish swine herds quarantined for pseudorabies in Illinois participated in the USDA-initiated Large Herd Cleanup Study. These herds were monitored for antibodies to pseudorabies virus (PRV) for 1 year after the initiation of an intensive eradication program. Herd size ranged between 425 and 1,500 females of breeding age. Gene-deleted modified-live virus vaccines were used on all farms, with 3 of the 6 herds receiving a vaccine with a deletion of the gene for glycoprotein-I and the other 3 herds receiving a vaccine with a deletion of the gene for glycoprotein-X. The breeding herd and growing pigs were vaccinated on each farm. Each herd produced its own replacement gilts. In addition, management changes emphasizing all-in, all-out pig flow were initiated. One year after initiation of the vaccination program, sera for the measurement of PRV antibodies were obtained from sows and heavy finishing pigs (> 70 kg) from each of the farms. Prevalence of PRV antibodies attributable to wild-type virus infection ranged from 7 to 63% (median, 33%) for sows and from 0 to 42% (median, 4%) for finishers, as determined by the appropriate vaccine differential test. For each sow herd, there was a large decrease in the PRV seroprevalence rate after 1 year of the program (range, -21 to -68%; median, -42%). Examination of PRV prevalence rates by parity indicated decreased seroprevalences in the lower parities (< 2) in 3 of the herds, suggesting that vaccination reduced the spread of PRV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407444 TI - Fish and seafood inspection a must. PMID- 8407445 TI - Development of hydrometra in a ewe flock after ultrasonography for determination of pregnancy. AB - Hydrometra was diagnosed in 41 of 1,411 (2.9%) Rambouillet and crossbred Rambouillet-Booroola Merino range ewes that were being examined with ultrasonography for pregnancy. Most diagnoses of hydrometra (40 of 41) were made at a second examination of 581 of the 1,411 ewes 40 days after the first examination (6.9%). Some of the ewes had membranes or small placentomes evident in the uterine fluid, suggesting that there had been embryonic resorption. Of the 15 ewes with hydrometra that were examined ultrasonographically a third time after an additional 42 days, 14 had resolved the condition. The increased prevalence of hydrometra at the second examination suggested that the stress of the first examination may have been a factor. Ovine fetal loss has been documented via ultrasonography, but hydrometra has not been commonly observed. Hydrometra may have developed because of unique circumstances of this flock, or it may be a condition that goes largely undiagnosed because most ewes are not examined for pregnancy twice within a short period. PMID- 8407446 TI - Prevalence of shigellosis and other enteric pathogens in a zoologic collection of primates. AB - An epidemiologic study of shigellosis was the preliminary step in the formulation of a plan for the control of devastating infectious diseases in nonhuman primates at the National Zoological Park. Data were collected from primate groups with enzootic shigellosis and included the following species: white-cheeked and siamong gibbons (Hylobates concolor and H syndactylies); lion-tailed, celebes, and Barbary macaques (Macaca silenus, M nigera, and M sylvanus); black and white colobus monkeys (Colobus guerzea); grey-cheeked mangabeys (Cerecocebus albigena); spider monkeys (Ateles susciceps robusuts); ruffed lemurs (Lemur varrigatus); lowland gorillas (Gorilla gorilla); and orangutans (pongo pygmaeus). Data included results of physical examination, proctoscopy with biopsy, fecal parasitologic and cytologic examinations, and bacteriologic culturing of swabbed specimens of rectum and gingiva. Repetitive fecal examinations were subsequently performed and included bacteriologic culturing of fecal specimens for enteropathogenic bacteria and parasites and cytologic examination of feces. Data were collected for a 1-year period from 82 primates, and 14 gibbons were studied intensively. White-cheeked and siamang gibbons shed Shigella flexneri sporadically, but persistently. All gibbons were affected with a mean point prevalence of 30.7% (range 0 to 71%). Shigella flexneri also was isolated from feces of lion-tailed macaques. Shigella sonnei was isolated from feces of grey cheeked mangabeys, celebes macaques, and spider monkeys.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407447 TI - Clearance of Shigella flexneri carriers in a zoologic collection of primates. AB - In 1990, a high prevalence of Shigella flexneri was documented in the gibbon population at the National Zoological Park. The enzootic strain had been determined to be resistant to most antibiotics and had been responsible for animal deaths. Prevalence of Shigella spp was high in the entire primate population. To address the clinical problem, eradication was attempted by use of enrofloxacin. Facilities disinfection comprised a large part of Shigella clearance efforts. After the clearance protocol was completed gibbons, vs other primate species, were closely monitored for 16 months by subjecting their feces to bacteriologic culturing. The program was successful, with eradication of the problematic strain of S flexneri. However, 10 to 12 months after completion of treatments, S sonnei was isolated from feces of 3 animals living in the same housing facility. PMID- 8407448 TI - When an itch means an allergy. PMID- 8407449 TI - Over-the-counter sale of rabies vaccine. PMID- 8407450 TI - Sustainable agriculture. PMID- 8407451 TI - Animal research. PMID- 8407452 TI - Recording vaccination sites. PMID- 8407453 TI - Dental prophylaxis not routine. PMID- 8407454 TI - United States Air Force Veterinary Corps: 1949 to 1980. PMID- 8407455 TI - A fulfilling career in academia. PMID- 8407456 TI - Anesthetic and medical management of acute hemorrhage during surgery. PMID- 8407457 TI - What is your diagnosis? Congenital fusion of vertebrae C1 and C2. PMID- 8407458 TI - What is your neurologic diagnosis? Infarction of spinal cord caused by fibrocartilaginous embolus. PMID- 8407459 TI - ECG of the month. PMID- 8407460 TI - Amitraz toxicosis associated with ingestion of an acaricide collar in a dog. PMID- 8407461 TI - Case of the month. Maternal behavior toward an unrelated kitten and lactation in an ovariohysterectomized cat. PMID- 8407462 TI - Injury by animals: a veterinarian's liability. PMID- 8407463 TI - AVMA voices profession's concerns over lay purchase/use of rabies vaccines. PMID- 8407464 TI - Radiographic anatomy and technique for arthrography of the cubital joint in clinically normal dogs. AB - A technique for arthrography of the cubital joint in clinically normal large breed dogs was developed with the objective of improving visualization of the articular margin of the medial coronoid process. A lateral approach to the cubital joint for injection of contrast medium was selected. Arthrography of 24 cubital joints was performed by using 14 dogs. Twelve combinations of iodinated contrast medium, consisting of various concentrations (3) and volumes (4), were used. Two sets of arthrograms for each of the 12 combinations of contrast medium were obtained. Five radiographic views were used for each set. All arthrograms were examined by 3 evaluators, and each articular surface received a numerical rating for how well it could be seen in each view. Results of the evaluation indicated that low volumes of contrast medium were preferable to high volumes, with 2 ml providing the best visualization. Concentration of iodine seemed less important than did volume. The numerical ratings also indicated that the articular margin of the coronoid process was clearly observed a maximum of only 24% of the time on a slightly supinated mediolateral projection. The articular margins of the head of the radius, trochlea humeri, and trochlear notch were well visualized > 90% of the time. Arthrography of the cubital joint was technically easy to perform, and complications were not encountered, but arthrographic anatomy of the cubital joint is complex. Potential uses for arthrography of the cubital joint include diagnosis of osteochondrosis, intraarticular fragments, and joint capsule ruptures. PMID- 8407465 TI - Comparison of radiologic imaging techniques for diagnosis of fragmented medial coronoid process of the cubital joint in dogs. AB - Thirty cubital joints from 16 dogs suspected of having a fragmented medial coronoid process were examined. Four breeds accounted for 87.5% of the cases: German Shepherd Dog (25%), Labrador Retriever (25%), Rottweiler (18.75%), and Golden Retriever (18.75%). Seventy-five percent of the dogs were male. Mean age of affected dogs was 13.6 months. Plain-film radiography, xeroradiography, linear tomography, arthrography, and computed tomography were performed on each cubital joint prior to surgical exploration of the joint. Three reviewers evaluated each diagnostic study and independently determined whether a fragment from the medial coronoid process could be seen. The consensus opinion was compared with the finding at surgery. Abnormalities of the medial coronoid process were detected in 25 of 30 joints at surgery. Fragmented coronoid process was found in 17 of 30 joints, and wear lesions were observed in 8 of 30 joints. Computed tomography had the highest accuracy (86.7%), sensitivity (88.2%), and negative-predictive value (84.6%) of the 5 imaging modalities evaluated (P < 0.05). Specificity and positive-predictive value of all imaging techniques were high. There was no significant difference between the diagnostic ability of plain-film radiography, xeroradiography, or linear tomography of the cubital joint. The combination of plain-film radiography and linear tomography provided an improvement in accuracy, approaching that of computed tomography. PMID- 8407466 TI - Comparison of the diagnostic accuracy of positive-contrast arthrography and arthrotomy in evaluation of osteochondrosis lesions in the scapulohumeral joint in dogs. AB - During a 6-year-period (1985-1991), 120 positive-contrast arthrograms were made in 70 dogs to evaluate osteochondrosis lesions in the scapulohumeral joint. In 76 shoulders, the arthrographic findings were compared with surgical or histopathologic findings to evaluate the accuracy of the arthrographic technique. Positive-contrast arthrography was found to be accurate in evaluation of the status of the articular cartilage. In revealing discontinuity of the articular cartilage, the results of arthrography were false-negative in 4 shoulders (58/66; accuracy, 88%). In detecting thick cartilage covering a subchondral defect, the results of positive-contrast arthrography were false-positive in 4 joints (6/11; accuracy, 55%). In detecting fragmentation of the cartilage, arthrography provided false-negative results in 4 joints (7/11; accuracy, 64%). This technique enabled detection of joint mice, especially within the bicipital tendon sheath (no false-positive results). Arthrography failed to reveal joint mice within the caudal pouch in 2 joints (15/17; accuracy, 88%). Joint effusion also could be evaluated and was correlated with synovitis, degenerative joint disease, and loose or detached cartilage flaps. Postarthrography complications were not observed with this technique. PMID- 8407467 TI - Choroidal melanoma in a dog. AB - B-scan ultrasonography of the right eye of a 6-year-old sexually intact female mixed-breed dog revealed complete retinal detachment and a semiconical solid mass of tissue that projected from the area of the optic disc into the vitreal space subretinally. Microscopic examination of an ultrasound guided fine-needle aspirate of the mass revealed a moderate number of mildly pleomorphic melanocytes. Histologic examination of the excised globe revealed a large, heavily pigmented intraocular neoplastic mass in the choroid around the optic nerve, bulging into the subretinal space. The histologic diagnosis was benign choroidal melanoma, with invasion into the intraocular portion of the optic nerve. In dogs, tumors of melanocytic origin are the most common primary ocular neoplasm, but primary choroidal melanocytic neoplasms are the rarest of this type. PMID- 8407468 TI - Urolithiasis in Dalmations: 275 cases (1981-1990). AB - From July 1, 1981 to December 31, 1990 the Urinary Stone Analysis Laboratory, School of Veterinary Medicine, University of California received 292 urinary calculi from 275 Dalmations (14 females, 14 episodes; 261 males, 278 episodes). The mean age of dogs at the time of the first episode of calculus formation was 4.5 years for males and 5.5 years for females. Bacteria were isolated from 22 of 49 calculi (45%). The bacteria were gram-positive with 2 exceptions; coagulase positive staphylococci accounted for 62.5% of the isolates. Polarized light microscopy was used to determine the mineral content of the calculi. One hundred ninety three calculi were 100% urate; 78 calculi contained > or = 50% urate in 1 or more layers and were classified as mixed urate calculi; 7 calculi contained < 50% urate in all layers; and 14 calculi contained no urate. The secondary minerals most commonly encountered in mixed urate calculi were struvite (77%, 60 of 78 calculi), and oxalate (17%, 13 of 78 calculi). There were 65 struvite containing calculi, 16 oxalate-containing calculi, 7 apatite-containing calculi, 5 silica-containing calculi, and 1 calculus specimen was composed of 100% cystine. The risk of forming urate-containing calculi was high in Dalmations, whereas the risk of forming calculi containing other minerals was consistently lower in Dalmations than in other breeds. X-ray diffraction and high pressure liquid chromatography were performed on all calculi from dogs known to have been given allopurinol (n = 19).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407469 TI - Autogenous biologics guidelines issued by AVMA. American Veterinary Medical Association. PMID- 8407470 TI - Conservation and wildlife preservation. PMID- 8407471 TI - Concerns about bovine somatotropin. PMID- 8407472 TI - Concerns about bovine somatotropin. PMID- 8407473 TI - "Rigorous standards of scholarship and creativity" needed. PMID- 8407474 TI - Animal research. PMID- 8407475 TI - Safety of foods of animal origin. PMID- 8407476 TI - What is an adequate education for the food animal practitioner? PMID- 8407477 TI - Antibacterial use in lactating dairy cattle. PMID- 8407478 TI - What is your diagnosis? Abdominal mass. PMID- 8407479 TI - Difficult dermatologic diagnosis. PMID- 8407480 TI - Malignant catarrhal fever in a gaur (Bos gaurus) cow. PMID- 8407481 TI - Veterinary-practice client volume. PMID- 8407482 TI - Causes and costs of calf mortality in Colorado beef herds participating in the National Animal Health Monitoring System. AB - Seventy-three Colorado cow/calf operations were monitored for calf mortality from birth to weaning as part of their participation in the National Animal Health Monitoring System. Producer-observed causes of calf mortality, and the costs associated with these deaths were obtained. The overall calf mortality during the study was 4.5%, with a total associated cost of $237,478. The mean cost per calf death was $216, of which $208 was attributed to the potential value of the calf and an additional $8 was for veterinary, drug, producer's labor, and carcass disposal expenses. The most commonly reported causes of calf mortality were dystocia (17.5%), stillbirth (12.4%), hypothermia (12.2%), diarrhea (11.5%), and respiratory infections (7.6%). These 5 disease conditions accounted for > 60% of all calf deaths. A cause was not determined for 19.7% of the calf deaths. Beef producers and veterinarians have the potential to decrease calf mortality and increase profits in cow/calf operations by implementing management strategies and herd health programs designed to decrease the number of calf deaths caused by these disease conditions. PMID- 8407483 TI - Bovine immunization guidelines. Council on Biologic and Therapeutic Agents, American Veterinary Medical Association. PMID- 8407484 TI - Effectiveness of buspirone on urine spraying and inappropriate urination in cats. AB - The most frequent type of behavior problem in cats for which veterinary consultation is sought is problem urination. Urine spraying and urine marking have been treated by use of long-acting progestins and diazepam, a benzodiazepine antianxiety drug. Effectiveness of the nonbenzodiazepine antianxiety drug, buspirone, in suppressing urine spraying and marking in 47 male and 15 female cats was evaluated. The effect of the drug in correcting inappropriate urination in 9 cats also was evaluated. Buspirone resulted in a favorable response (> 75% reduction) in 55% of cats treated for urine spraying or marking. There was no sex difference in effectiveness of the treatment, but cats from single-cat households responded favorably significantly (P < 0.001) less frequently than those from multiple-cat households. The 55% response rate was within the range of treatment effectiveness that has been reported for diazepam, and was greater than that reported for progestin. In contrast to diazepam, with which over 90% of treated cats resumed spraying or marking when the drug was gradually discontinued, only half of the cats treated with buspirone resumed spraying when the drug was discontinued after 2 months of treatment (P < 0.001). This difference between diazepam and buspirone in resumption of urine spraying was attributed to diazepam's induction of physiologic and behavioral dependency, not found with buspirone. Cats that resumed spraying were placed on long-term treatment ranging from 6 to 18 months. Buspirone also did not cause the adverse effects of sedation and ataxia, which commonly are seen with diazepam treatment. In cats treated for inappropriate urination, 56% returned to normal litter box usage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407485 TI - Diagnosis of food allergy in dogs. AB - The diagnosis of food allergy was confirmed in 51 dogs while the responsiveness to a 60-day home-cooked restricted dietary trial (elimination-diet trial) was evaluated. The primary clinical sign of allergy detected and evaluated in all dogs was persistent and nonseasonally pruritic skin disease. The duration of time between starting the elimination-diet trial and remission of clinical signs was recorded. Dogs were then reexposed to diets that had been fed before testing, and the duration of time before pruritus recurred was recorded. The elapsed time during which dogs were being fed an elimination diet before remission of clinical signs was 1 to 3 weeks in 13 dogs, 4 to 6 weeks in 25 dogs, 7 to 8 weeks in 10 dogs, and 9 to 10 weeks in 3 dogs. Findings indicated that the recommendation of a 3-week elimination-diet trial for diagnosis of food allergy was adequate for only 25% of the dogs. It is recommended that test diets be fed for at least 10 weeks before a food allergy is ruled out. PMID- 8407486 TI - Subcutaneous Taenia crassiceps cysticercosis in a dog. AB - Taenia crassiceps larvae, typically found in rodents, were recovered from fluctuant subcutaneous cystic lesions in a dog. Differential diagnosis included Taenia crassiceps cysticercosis, other metacestode infections, and pseudotumoral skin diseases. PMID- 8407487 TI - Supernumerary tooth associated with facial swelling in a dog. AB - Facial swelling in a dog was found to be associated with a supernumerary premolar. Extraction of the tooth alleviated the swelling. Although supernumerary teeth are common, they are not known to be associated with facial swelling. PMID- 8407488 TI - Unilateral adrenalectomy as a treatment for adrenocortical tumors in ferrets: five cases (1990-1992). AB - Adrenocortical tumors were diagnosed in 5 adult spayed ferrets. Four ferrets had bilaterally symmetrical alopecia of the caudal femoral region, abdomen, and tail, and 1 had alopecia of the distal limbs and feet. All 5 ferrets had vulvar swelling. During abdominal ultrasonography, irregular masses, believed to involve the adrenal glands, were seen in all 5 ferrets. Unilateral adrenalectomy was performed successfully in each ferret by use of ventral midline celiotomy. On histologic examination of biopsy samples, 4 ferrets were found to have adrenocortical adenomas, and 1 ferret was found to have an adrenocortical adenocarcinoma. All clinical signs resolved after adrenalectomy, suggesting that the adrenocortical tumors had been secreting adrenocortical hormones. PMID- 8407489 TI - Hyperadrenocorticism associated with adrenocortical tumor or nodular hyperplasia of the adrenal gland in ferrets: 50 cases (1987-1991). AB - Adrenocortical adenoma, nodular hyperplasia, or carcinoma was diagnosed in 50 ferrets. Thirty-five (70%) ferrets were female and 15 (30%) were male. The mean age at which clinical signs were first noticed was 3.4 years (range, 1 to 7 years). Clinical signs included large vulva (n = 31; 89% of females), alopecia (n = 43; 86%), pruritus (n = 20; 40%), and increased consumption of water and increased urine output (n = 4; 8%). A mass was palpated at the cranial pole of the kidney during physical examination of 17 (34%) ferrets. Ultrasonography, performed on 39 of 50 ferrets, revealed a unilateral adrenal gland mass in 19 (49%). Four ferrets were anemic, and 2 ferrets were thrombocytopenic. Baseline plasma concentrations of cortisol and corticosterone were within or below the reference range in all 17 ferrets tested, whereas baseline plasma estradiol concentrations were high in 4 of the 11 ferrets (36%) tested. AFter adrenocorticotropic hormone (ACTH) administration, only 1 ferret had a slightly exaggerated response on the basis of plasma cortisol concentrations, and all 17 had normal responses on the basis of plasma corticosterone concentrations. There was little or no increase in plasma estradiol concentrations after ACTH administration. Of the 50 ferrets, 39 were treated by adrenalectomy. Unilateral adrenalectomy was performed in 34 ferrets in which 1 adrenal gland was large, whereas subtotal bilateral adrenalectomy was performed in 5 ferrets with bilateral adrenal disease. Five ferrets died in the immediate postoperative period, and follow-up information was available for the remaining 34, 1 to 34 months after surgery.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407490 TI - Hematuria and leptospiruria in a foal. AB - A Thoroughbred foal that was born after 305 days of gestation was referred 6 hours after birth. On initial examination, the foal was alert, but weak and unable to stand. Heart rate was 150 beats/min, respiratory rate was 48 breaths/min, and rectal temperature was 33 C. Leukocytosis was detected (26,000 WBC/microliters). Dysuria and passage of red urine was observed approximately 30 hours after admission. Urine collected by free catch had a 3+ reaction for blood on dipstick analysis. Examination of Warthin Starry-stained sections of allantochorion revealed organisms morphologically characteristic of leptospira spp along the chorion. Results of direct fluorescent antibody tests on section of placenta were positive for Leptospira spp. Direct fluorescent antibody testing identified leptospires in the urine of the foal. Sera from mare and foal were tested for antibodies against Leptospira spp by use of the microscopic agglutination test. High titers were identified in serum from the mare and foal. PMID- 8407491 TI - Standing laparoscopic laser technique for ovariectomy in five mares. AB - Laparoscopic laser instrumentation and endoscopic stapling techniques were used to perform 10 unilateral ovariectomy procedures on 5 standing Thoroughbred mares. In all instances, mares had a clinically normal reproductive tract and estrous cycle, and ovariectomy was performed to eliminate regular estrous behavior. All procedures were accompanied by minimal trauma and uncomplicated postoperative recovery. Mares were able to resume normal activity 2 weeks after surgery. Regular estrous behavior was eliminated in all 5 mares, although 1 mare continued to have signs of estrous behavior occasionally after the bilateral ovariectomy. Advantages of standing laparoscopic laser ovariectomy over open laparotomy or colpotomy techniques included minimal surgical morbidity, decreased postoperative discomfort, and rapid, uncomplicated healing. Disadvantages of laparoscopic laser ovariectomy included cost of specialized equipment, need for advanced training in laser and laparoscopic surgery, and increased operative time. PMID- 8407492 TI - Peritonitis in horses: 67 cases (1985-1990). AB - Peritonitis was diagnosed in 67 horses between 1985 and 1990: 14 horses developed septic peritonitis after intestinal rupture, 25 horses developed peritonitis after abdominal surgery, and 28 horses had peritonitis not associated with intestinal rupture or abdominal surgery. Forty of 67 horses (59.7%) did not survive. Nonsurvivors had higher heart rates (P = 0.01), RBC count (P = 0.039), serum creatinine concentration (P = 0.036), PCV (P = 0.007), and anion gap (P = 0.005); lower venous blood pH (P = 0.002); and a greater number of bacterial species cultured from peritoneal fluid samples (P = 0.054), compared with those from survivors. Nonsurvivors were more likely to have signs of abdominal pain (P < 0.000), circulatory shock (P = 0.009), and bacteria in peritoneal fluid samples (P = 0.042). Physical examination and peritoneal fluid analysis were the most valuable diagnostic aids for intestinal rupture. Peritonitis after abdominal surgery resulted in high mortality (56%); peritonitis not associated with intestinal rupture or abdominal surgery had lower mortality (42.9%). Clinical and laboratory indices can be of value in determining the prognosis for horses with peritonitis. PMID- 8407493 TI - Transmission of brucellosis from reindeer to cattle. AB - Sixteen reindeer (Rangifer tarandus tarandus) naturally infected with Brucella suis biovar 4 were penned with 6 male and 2 female cattle for 30 days, then removed and euthanatized. During this period, 5 reindeer had fawns, and 2 reindeer aborted. Brucella suis biovar 4 was recovered from all adult reindeer at necropsy. Nine reindeer had B suis biovar 4 in uterus, udder, and/or milk. The cattle were euthanatized 2 months after the reindeer were removed. Clinical or pathologic signs of disease were not observed in the cattle. Brucella suis biovar 4 was isolated from 2 male and from both female cattle at necropsy. The female cattle had positive reactions on the buffered plate antigen test, brucellosis card test, tube agglutination test, complement fixation test, and indirect enzyme immunoassay for most of the experiment, but the males had inconsistent reactions on these tests. The indirect enzyme immunoassay was the only test to detect all cattle from which bacteria were cultured. This study revealed that caution is warranted before moving reindeer or caribou into areas of traditional agriculture. PMID- 8407494 TI - Effect of management practices on the Streptococcus suis carrier rate in nursery swine. AB - Management practices on swine farms were analyzed to determine factor(s) associated with high prevalence of pigs that were carriers of Streptococcus suis. Samples were obtained for bacteriologic culture via direct swabbing of palatine tonsils of healthy nursery pigs on 35 farms throughout the United States. Overall, 36.7% of the pigs were determined to be carriers. Isolates of S suis were serotyped, and antimicrobial susceptibility tests were performed by use of Kirby-Bauer techniques. Streptococcus suis types 1 and 2 were most commonly isolated. All isolates were susceptible to enrofloxacin, 97% of the isolates were susceptible to ceftiofur, and 94% were susceptible to ampicillin. However, only 80% of the isolates were susceptible to penicillin, and only 18% were susceptible to tetracycline. Environmental, managerial, nutritional, and health factors were measured on each farm. Excessive temperature fluctuation, high relative humidity, crowding, and an age spread of > 2 weeks between pigs in the same room were the 4 most commonly encountered problems on farms with higher-than-average percentages of carrier pigs. Continuous flow facilities were found on 50% of these farms, and various disease problems, vitamin E/selenium deficiency, inadequate vaccination programs (attributable to the presence of atypical serotypes), and penicillin resistant strains were found on 6 to 28% of these farms. Overall, 83% (15/18) of farms with higher-than-average percentages of carrier pigs also had a history of clinical S suis disease. PMID- 8407495 TI - Endothelium and inflammation. PMID- 8407496 TI - Keep abreast of changes in Pasteurized Milk Ordinance. PMID- 8407497 TI - Socialism to market economy. US eases growing pains of Latvian veterinary profession. PMID- 8407498 TI - Ophthalmologists need to focus on extra-label drug use initiative. PMID- 8407499 TI - Mandatory sterilization of dogs and cats. PMID- 8407500 TI - Unlicensed practice. PMID- 8407501 TI - The use of animals in testing: a focal point for controversy. PMID- 8407502 TI - Integrated resource management. PMID- 8407503 TI - Our community and us. PMID- 8407504 TI - What is your diagnosis? Hepatomegaly, abdominal mass and multiple gas emboli. PMID- 8407505 TI - Diagnosis and treatment of closed-cervix pyometra in a bitch. PMID- 8407506 TI - Veterinary practice financial loans. PMID- 8407507 TI - Incidence and economics of tuberculosis in swine slaughtered from 1976 to 1988. AB - The Swine Tuberculosis Regulations, revised in 1972, stipulated that all swine carcasses with mycobacterial lesions in more than 2 primary sites should be passed for cooking (PFC). Economic loss from a condemned carcass is 100%, whereas loss from a PFC carcass is 66%. Increased condemned and PFC swine carcass rates in 1972, 1973, and 1974, and the economic losses from them were attributed to changes in the regulations. An industrial organization estimated increased economic losses from swine tuberculosis, but detected decreased rates of condemned and PFC swine carcasses in 1975 and 1986. The federal meat inspection data for 1976 to 1988 indicated that the yearly condemned carcass rate remained < 8.0/100,000 swine slaughtered, whereas the PFC carcass rate decreased by 74.1%, from 52.4 to 13.6/100,000 swine slaughtered. The incidence (condemned + PFC) per 100,000 swine slaughtered decreased by 67.7%, from 58.03 in 1976 to 18.72 in 1988. The Agricultural Statistics indicated that a yearly loss from tuberculosis of $2.3 million in 1976 decreased by 73% to $0.97 million in 1988. A yearly loss of $41,580/$100 million of animal value decreased by 70% to $12,880/$100 million in 1988. The decreased incidence of swine tuberculosis and the economic losses with this disease indicate that the swine industry in the United States was not adversely affected by the change in the Swine Tuberculosis Regulations. PMID- 8407508 TI - Comparison of the efficacy of cefadroxil and generic and proprietary cephalexin in the treatment of pyoderma in dogs. AB - We compared efficacy of cefadroxil and generic and proprietary cephalexin in treatment of pyoderma in dogs. Forty-four dogs were randomly assigned to receive 1 of the 3 preparations at 22 to 35 mg/kg body weight, every 12 hours. Dogs were examined at the conclusion of treatment period and assessed as to degree of improvement. All 3 cephalosporins were effective and safe antibiotics for the treatment of pyoderma in dogs. PMID- 8407509 TI - Assessment of corticosteroid-induced alkaline phosphatase isoenzyme as a screening test for hyperadrenocorticism in dogs. AB - Quantitative determination of the corticosteroid-induced isoenzyme of alkaline phosphatase (CAP) was evaluated as a screening test for hyperadrenocorticism (HAC) in dogs. A series of 40 dogs with HAC (CAP range, 96 to 14,872 U/L), 30 clinically normal dogs (CAP range, 0 to 38 U/L), and 80 dogs with various diseases (non-HAC) and without history of exogenous glucocorticoid exposure for a minimum of 60 days (CAP range, 0 to 1163 U/L) were used to evaluate the test. Sensitivity and specificity of CAP was calculated at various cutoff points for absolute CAP activity and for CAP activity expressed as a percentage of total alkaline phosphatase activity. A cutoff point of 90 U/L was selected as optimal for use of this assay as a screening test for HAC. A prevalence survey then was done of all canine serum samples submitted to our diagnostic laboratory over a 3 month period, to calculate the predictive values of a positive and a negative test result in a clinical population and to determine the relative frequency and magnitude of CAP activity in dogs that had received glucocorticoids. The predictive values of a positive and a negative test result at the 90 U/L cutoff value were 21.43% (95% confidence limits, 8.3 to 40.95%) and 100% (95% confidence limit > 96%), respectively. It was concluded that CAP isoenzyme activity, determined by routine biochemical analysis by an automated levamisole-inhibition assay, could function as a screening test for HAC; however, the predictive value of a positive test result was too low to recommend the assay as a diagnostic test.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407510 TI - Spinal ganglioneuroma in a dog. AB - A 10-year-old mixed-breed dog was examined because of lameness and mutilation of the left hind limb. Neurologic examination revealed ataxia, conscious proprioceptive deficit, muscle atrophy, lack of withdrawal reflex, and hyperreflexia of the patella of the left hind limb. Epidurography revealed right sided deviation of the dye column at the sixth to seventh lumbar vertebrae. Laminectomy with cytoreduction was performed, and histologic examination revealed ganglioneuroma. Twelve months after surgery, clinical signs recurred and surgery was repeated. The dog's clinical signs were again palliated. The specific type of ganglioneuroma identified in this dog is amenable to surgical resection, unlike many types of spinal cord neoplasia. PMID- 8407511 TI - Adverse drug reaction to metoclopramide hydrochloride in a macaw with proventricular dilatation syndrome. AB - A 4-year-old female blue and gold macaw (Ara ararauna) with a history of chronic vomiting was treated with metoclopramide hydrochloride. After the second treatment, ataxia, torticollis, and opisthotonos became evident. These signs resolved with the administration of diphenhydramine hydrochloride. Despite supportive care, the bird died several days later. Histologic lesions were suggestive of proventricular dilatation syndrome. PMID- 8407512 TI - Uveal cysts in dogs: 28 cases (1989-1991). AB - In a 3-year retrospective study, 28 dogs were determined to have uveal cysts arising from either the ciliary body or the iris or free-floating in the anterior chamber. Golden Retrievers, Labrador Retrievers, and Boston Terriers were represented more than other breeds. Mean age of cyst development was 7.0 years in Golden Retrievers, 9.1 years in Labrador Retrievers, and 6.8 years in Boston Terriers. There was no sex predilection in any breed. Evidence of associated ocular lesions, other than visual impairment in 1 Boston Terrier, was not noticed. PMID- 8407513 TI - Endoparasitism in dogs: 21,583 cases (1981-1990). AB - Prevalence of endoparasites (particularly ascarids) in dogs examined at the Oklahoma State University veterinary medical teaching hospital during 1981 to 1990 was determined by fecal and blood examinations. Approximately 1,250 fecal and 900 blood specimens were examined each year. In 1981, 55% of dogs harbored 1 or more parasites, compared with 36% in 1990. Percentages of all endoparasitic species decreased, except Giardia spp, which increased. The greatest decrease in prevalence was for Ancylostoma spp, which changed from 39% in 1981 to 15% in 1990. Prevalence of Toxocara spp and Trichuris spp also significantly decreased from 1981 (8% and 12%, respectively) through 1990 (4% and 9%, respectively). Although there appeared to be a downward trend in these ascarid infections, considerable environmental contamination probably existed and continues to exist because of high fecundity and long survival of eggs in the environment. Prevalence of Dipetalonema spp had a downward trend from a high of 4% in 1981 to < 1% by 1991. Prevalence of Dirofilaria spp decreased from 12% in 1982 to 6% in 1985, but by 1990, it was 11%. PMID- 8407514 TI - Disseminated Halicephalobus deletrix infection in a horse. AB - A 13-year-old Quarter Horse gelding was referred for evaluation of a draining tract and fracture of the right hemimandible of 4 weeks' duration. Two days prior to admission, the horse had developed pigmenturia. Radiography of the mandible revealed a fracture of the vertical ramus of the right hemimandible, loss of the right lower second premolar, and osteomyelitis involving an extensive portion of the hemimandible. Ultrasonography of the left kidney revealed loss of normal renal architecture. Histologic examination of tissue obtained from the right hemimandible revealed granulomatous osteomyelitis and multiple metazoan parasites identified as Halicephalobus deletrix. The horse was treated with antimicrobial and deworming agents, and the mandible was surgically debrided. The horse became atactic 8 days after surgery and was euthanatized. Necropsy identified parasitic migration in the right hemimandible, kidneys, and CNS. PMID- 8407515 TI - Removal of a fracture fragment from the palmar aspect of the intermediate carpal bone in a horse. AB - A 10-year-old Dutch Warmblood gelding sustained a fracture of the palmar tuberosity of the intermediate carpal bone 2 weeks prior to admission. The dorsolateral-palmaromedial oblique and flexed lateromedial radiographic views revealed the fracture line. The palmar fracture fragment from the intermediate carpal bone was removed by arthrotomy of the palmar intercarpal joint, which was approached through the carpal canal. Twenty months after surgery, the horse was show jumping without evidence of lameness. PMID- 8407516 TI - Seminal vesiculitis as a cause of signs of colic in a stallion. AB - A 5-year-old stallion was referred because of signs of abdominal pain. During the initial examination, signs of pain were elicited when the right seminal vesicle was palpated per rectum. Signs of pain were also elicited during sexual arousal and attempts at semen collection. The right seminal vesicle was subsequently determined to be abnormal by ultrasonographic and endoscopic examination. The stallion was treated with trimethoprim/sulfamethoxazole for 6 weeks. Five months later, there had been no recurrence of the condition. PMID- 8407517 TI - Spaying/neutering comes of age. PMID- 8407518 TI - Of mites and man. PMID- 8407519 TI - Nonveterinarian ownership of practice facilities. PMID- 8407520 TI - Diagnosing canine borreliosis. PMID- 8407521 TI - Care of research animals. PMID- 8407522 TI - Care of research animals. PMID- 8407523 TI - Research on Borrelia burgdorferi. PMID- 8407524 TI - Ecotoxicology and ecosystem health: roles for veterinarians; goals of the Envirovet program. PMID- 8407525 TI - Potlach: a philosophy worth living. PMID- 8407526 TI - Ostrich chick survival presents challenge. PMID- 8407527 TI - What is your diagnosis? Interruption of the aortic arch and subaortic stenosis in a dog. PMID- 8407528 TI - What is your neurologic diagnosis? Degenerative myelopathy in a dog, based on negative results of all diagnostic tests. PMID- 8407529 TI - ECG of the month. PMID- 8407530 TI - Animal behavior case of the month. PMID- 8407531 TI - Veterinarians and minors. PMID- 8407532 TI - Nonsurgical removal of urocystoliths in dogs and cats by voiding urohydropropulsion. AB - A technique called voiding urohydropropulsion has been developed that facilitates nonsurgical removal of urocystoliths. Voiding urohydropropulsion was performed in 11 dogs and 10 cats with urocystoliths. Urocystoliths were completely removed from 15 of 21 animals (5 female dogs, 3 male dogs, 5 female cats, and 1 male cat). The number of uroliths removed from any animal varied between 1 and 983. The mean time required to complete voiding urohydropropulsion in the 15 animals from which all uroliths were completely removed was 22 minutes. In 6 animals (2 female dogs, 3 female cats, and 1 male cat), not all urocystoliths were removed. Visible hematuria was induced in all animals as a consequence of voiding urohydropropulsion. In dogs, visible hematuria resolved within 4 hours. Dysuria was not induced by this technique in dogs. In many cats, visible hematuria and dysuria persisted for 1 to 2 days. One male cat developed urethral obstruction after we failed to remove a urolith from the bladder. The urolith was returned to the urinary bladder, and subsequently removed by cystotomy. Voiding urohydropropulsion is a simple and effective method that should be considered for removal of small urocystoliths from dogs and cats before cystotomy is performed. PMID- 8407533 TI - Comparison of pentobarbital alone and pentobarbital in combination with lidocaine for euthanasia of dogs. AB - Pentobarbital alone, pentobarbital plus 1% lidocaine solution, pentobarbital plus 2% lidocaine solution, and pentobarbital plus 3% lidocaine solution were each used to euthanatize 6 dogs. For each dog, time between the beginning of injection of the euthanasia solution and each of the following events was recorded: collapse, onset of apnea, flat-line electrocardiogram, flat-line electroencephalogram, loss of palpable heartbeat, and loss of palpable pulse. Any signs of pain or discomfort were also recorded. There were no significant differences among groups except for time to flat-line electrocardiogram. Dogs euthanatized with pentobarbital alone had significantly longer times than did dogs euthanatized with pentobarbital in combination with any of the lidocaine concentrations. We concluded that pentobarbital in combination with lidocaine was a reasonable alternative to pentobarbital alone when euthanatizing dogs. PMID- 8407534 TI - Ovarian abscesses and pyometra in a domestic rabbit. AB - Infection with Pasteurella multocida can induce pyoendometritis, pyosalpingitis, and ovarian abscesses in rabbits. The likelihood of rabbits developing clinical pasteurellosis is predisposed by factors such as buildup of ammonia fumes, ambient temperature changes and drafts, reproduction, older age, existence of carriers, and poor sanitation. Traditionally, prevention of pasteurellosis in rabbits has focused on identifying suspected carriers and culling those animals from the rabbitry. PMID- 8407535 TI - Lymphoplasmacytic keratitis in a ferret with lymphoma. AB - Unilateral lymphoplasmacytic keratitis was believed to be associated with multicentric lymphoma in a ferret. Plasmacytosis (Aleutian disease) should be considered in any ferret with lymphoplasmacytic infiltration of any organ. Ferrets with antibodies to the parvovirus causing plasmacytosis are believed to be more susceptible to concurrent disease because of immunosuppression. PMID- 8407536 TI - Risk factors associated with acute pancreatitis in dogs: 101 cases (1985-1990). AB - The medical records of 101 dogs with acute pancreatitis, diagnosed on the basis of medical histories of acute vomiting, with serum lipase or amylase activity greater than the reference range, or with gross signs of pancreatitis at surgery or histopathologic evidence at necropsy, were evaluated to identify potential risk factors for the development of acute pancreatitis. Age, sex, and breed of dogs with acute pancreatitis were compared with those from a reference population of 100 dogs admitted for other medical emergencies during the same period. Analysis of multiple regression models indicated that dogs > 7 years old were at increased risk for acute pancreatitis. Spayed dogs and castrated male dogs had an increased risk, compared with that of sexually intact males. Similarly, terrier and nonsporting breeds appeared to be at higher risk of developing acute pancreatitis than were other breed types. Most dogs in this study (63/101) had intercurrent diseases, including diabetes mellitus (n = 14), hyperadrenocorticism (n = 12), chronic renal failure (n = 8), neoplasia (n = 17), congestive heart failure (n = 6), and autoimmune disorders (n = 5). Fourteen dogs had undergone anesthesia or surgery in the week before admission; only 3 had undergone abdominal procedures. Recent medication use was listed in 52 of 101 cases. Antibiotics (n = 18) and corticosteroids (n = 18) were most frequently described. Anticancer chemotherapeutic agents (n = 5) and organophosphate insecticides (n = 5) also were listed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407537 TI - Granulomatous hepatitis in dogs: nine cases (1987-1990). AB - Granulomatous hepatitis (GH) is an uncommon histopathologic diagnosis in dogs. On the basis of clinical reports, fungal infections appear to be the most common cause of GH in dogs, but many other potential causes have been identified. The medical records and histopathologic findings for 9 dogs with GH were reviewed to identify additional specific causes of GH in dogs. Diseases associated with GH included intestinal lymphangiectasia (n = 2), lymphosarcoma (n = 1), histiocytosis (n = 1), dirofilariasis (n = 1), and histoplasmosis (n = 1). In 1 dog, no other disease process was identified. Of the remaining 2 dogs, 1 had concurrent granulomatous pneumonitis of unknown cause, and the other had periportal hepatitis and temporal muscle wasting. All 9 dogs with GH had clinical evidence of liver disease, such as hepatomegaly, icterus, and ascites, or had high serum alkaline phosphatase and alanine aminotransferase activity. Because of the wide variety of potential causes of GH in dogs, an accurate diagnosis should be sought so that appropriate treatment can be chosen and an accurate prognosis given. PMID- 8407538 TI - Analysis of hemostasis in horses with colic. AB - Eight tests of hemostasis were measured in 233 horses with colic. Blood samples were obtained at admission and for 4 consecutive days of hospitalization. Data were analyzed retrospectively by outcome, by broad-category diagnosis group, by small intestinal disorder, and by smaller categories for comparing specific diseases. Nonsurviving horses and horses with the most severe forms of intestinal ischemia had changes interpreted as hypercoagulative, the intensity of which was increased on the first and second mornings (sample times 2 and 3) after admission, when most significant differences for results of specific tests were detected. Nonsurvivors had decreased antithrombin III activity and prolonged prothrombin and activated partial thromboplastin times; those with strangulating obstructions also had decreased protein C and plasminogen activities. During hospitalization and with survival, these changes tended to reverse. In most horses, regardless of diagnosis or outcome, concentration of fibrin degradation products and fibrinogen, and alpha 2-antiplasmin activity increased over time. Whether these changes reflected specific effects of colic or of the acute-phase response was not determined. In comparisons of small intestinal disorders (proximal enteritis, strangulations, and impactions), diagnostically distinguishing features were not found. Likewise, in comparisons of specific diseases (small vs large intestinal impaction, proximal enteritis vs colitis, small vs large intestinal obstruction), diagnostically distinguishing features were not found. PMID- 8407539 TI - Diffuse cerebral encephalopathy associated with hydrocephalus and cholesterinic granulomas in a horse. AB - Cholesterinic granulomas, commonly found at necropsy in aged horses, may cause neurologic signs by obstructing the interventricular foramina, resulting in hydrocephalus. Diffuse cerebral disease in horses may not always result in rapid progression of clinical signs. Intermittency of neurologic signs may be associated with intermittent increases in CSF pressure. PMID- 8407540 TI - Articular fracture of the dorsoproximolateral aspect of the third metatarsal bone in five standardbred racehorses. AB - Articular fracture of the dorsoproximolateral aspect of the third metatarsal bone (MT3) caused an acute onset of lameness in 5 horses; however, 3 of the horses had historical and radiographic or scintigraphic evidence of chronic tarsal and metatarsal disease before fracture. The pathogenesis of dorsoproximolateral fracture of MT3 remains unclear, but the fracture may occur as a result of the formation of abnormal bone in the proximal aspect of MT3. The prognosis for racing in horses with this fracture appears to be guarded. PMID- 8407541 TI - Use of orbital implants after enucleation in dogs, horses, and cats: 161 cases (1980-1990). AB - Eye enucleations performed on 109 dogs, 29 horses, and 23 cats involved placement of 136 silicone orbital implants and 7 mesh implants. Mean follow-up times were 2.4 years (range, 3 weeks to 9 years) in dogs, 3.4 years (range, 10 days to 10.5 years) in horses, and 1.5 years (range, 3 weeks to 7.5 years) in cats. Implants failed in 1 of 96 dogs (1.04%), 3 of 29 horses (10.3%), and 3 of 18 cats (16.7%). Implant failure was attributable to various causes in all species; however, cats appeared to be more prone to late extrusion that were dogs and horses. Implantation of an orbital prosthesis was a safe and inexpensive method for improving cosmetic appearance after enucleation in dogs, horses, and cats. PMID- 8407542 TI - Undercoat-constitutive proteins of cell-cell adherens junctions. PMID- 8407543 TI - Perspectives on cancer in Japan and the United States. PMID- 8407544 TI - HTLV-1-associated myelopathy/tropical spastic paraparesis-like rats by intravenous injection of HTLV-1-producing rabbit or human T-cell line into adult WKA rats. AB - We intravenously injected Ra-1 cells or MT-2 cells into female adult WKA rats. Spastic paraparesis mainly in the hind-limbs was observed in 1 out of 2 Ra-1 cell injected WKA rats and in 3 out of 8 MT-2 cell-injected WKA rats 20-27 months after injection. The main neuropathological finding was symmetrical white matter degeneration with mononuclear cell infiltration of the spinal cord, similar to that of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients, and degeneration of nerve roots and peripheral nerves. Antibodies against HTLV-1 antigens were detected in plasma and cerebrospinal fluid from these HAM/TSP-like rats. HTLV-1 provirus was detected from the peripheral blood mononuclear cells of one of these rats 20 months after injection. Interestingly, spastic paraparesis was not observed in F344 rats. PMID- 8407545 TI - Chemiluminescence immunoassay to detect anti-HTLV-I antibodies. AB - A fully automated chemiluminescence immunoassay was developed for the detection of antibodies to HTLV-I. We used partially purified viral antigens coated on small polystyrene beads together with acridinium ester-labeled anti-human immunoglobulin G mouse immunoglobulin G in this method. A good agreement was observed between our proposed method, the indirect immunofluorescence assay, the particle-agglutination test and the enzyme immunoassay. This new method, which is simple, sensitive, specific and rapid, should be useful for mass screening of anti-HTLV-I antibodies. PMID- 8407546 TI - Chromosomal mapping of genetic locus associated with thymus-size enlargement in BUF/Mna rats. AB - The thymoma-prone rat of the BUF/Mna strain is a useful model for human thymoma. In this strain thymoma development is regulated by a single autosomal susceptible gene, Tsr-1. At pre-thymoma age, BUF/Mna rats have extremely large thymuses, when compared to those of other strains of rats. Genetic studies in crosses between BUF/Mna rats with large thymuses and WKY/NCrj rats with small thymuses suggested the presence of a major autosomal gene, Ten-1, which contributes to thymus enlargement in a backcross population. Linkage studies between Ten-1 and microsatellite markers in backcross rats of (WKY/NCrj x BUF/Mna)F1 x BUF/Mna have led to the localization of Ten-1 in chromosome 1. This result may provide an approach to clone Tsr-1, which could be allelic to Ten-1. PMID- 8407547 TI - Suppression of rabbit VX-2 subcutaneous tumor growth by gadolinium neutron capture therapy. AB - VX-2 tumors growing in hind legs of New Zealand White rabbits (n = 4) were exposed to thermal neutrons for 40 min (2.1 x 10(12) neutrons cm-2) while one of two hind leg tumors of each rabbit was infused continuously with meglumine gadopentetate through a branch of the left femoral artery. The contralateral (uninfused) tumors served as controls. Although no differential distribution of gadolinium was achieved between the tumor and its adjacent normal tissue, the gadolinium concentration in the infused tumor was approximately 5-6 fold higher than that in the contralateral tumor. Growth of gadolinium-infused tumors was significantly inhibited compared to that of control tumors (P < 0.05) between the 16th and 23rd days after treatment. PMID- 8407548 TI - Does high gastric cancer risk associated with low serum ferritin level reflect achlorhydria? An examination via cross-sectional study. AB - With respect to the inverse association of serum ferritin level (SFL) with the risk of gastric cancer (GC) observed in some recent epidemiologic studies, possible mediation by achlorhydria as well as atrophic gastritis (AG), both of which are strongly associated with GC risk at not only the individual but also the population level, was examined in a cross-sectional study of 634 men aged 40 to 49 years randomly selected from 5 populations in Okinawa, Iwate, Nagano, Akita and Tokyo. AG and achlorhydria were serologically diagnosed based on the criteria of pepsinogen (Pep) I level < 70 ng/ml and Pep I/Pep II ratio < 3.0, as described previously, and a serum gastrin level of over 140 pg/ml, respectively. In the results, while the mean SFL for all the subjects differed significantly by area, similar areal differences in SFL were also found even when only the non-AG cases were considered. However, both of the above differences were eliminated with the exception of those between Okinawa and each of the other 4 areas, when adjustments were made for medical histories of diabetes mellitus, ulcers and liver disease, body mass index and gamma-glutamyltranspeptidase level. Therefore, no correlation among the 5 areas was observed between the adjusted areal mean SFLs and GC mortality in either case.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407549 TI - The rat urinary bladder as a new target of heterocyclic amine carcinogenicity: tumor induction by 3-amino-1-methyl-5H-pyrido[4,3-b]indole acetate. AB - In order to examine the carcinogenicity of 3-amino-1-methyl-5H-pyrido[4,3 b]indole acetate (Trp-P-2), 30 male and 30 female F344 rats were maintained on diet containing 0, 30, or 100 ppm Trp-P-2 for 112 weeks. The overall mean chemical intakes in the 100 ppm and 30 ppm groups were 3.84 and 1.14 mg/kg/day in males, and 4.57 and 1.34 mg/kg/day in females, respectively. Females of the 100 ppm group showed increased mortality in the late period of the study. In the 100 ppm group, significant increases in the incidences of neoplastic lesions were found in the liver, urinary bladder and mammary gland in males, and in the mammary gland, hematopoietic system and clitoral gland in females. Histologically, tumors induced by Trp-P-2 were hepatocellular adenomas, transitional cell tumors (papillomas and carcinomas) of the urinary bladder, fibroadenomas/fibromas of the mammary gland, malignant lymphomas and clitoral gland tumors (adenomas and adenocarcinomas). These results indicate multi-target carcinogenicity of Trp-P-2 in F344 rats and provide evidence that the urinary bladder is also a target for heterocyclic amine action. PMID- 8407550 TI - Effects of chronic administration of low doses of 2-amino-3,8-dimethylimidazo [4,5-f]quinoxaline on glutathione S-transferase placental form-positive foci development in the livers of rats fed a choline-deficient diet. AB - Effects of chronic administration of 2-amino-3,8-dimethylimidazo[4,5 f]quinoxaline (MeIQx) at the very low doses of 0.4 and 4 ppm, respectively 1000- and 100-fold less than the dose shown to be carcinogenic (400 ppm), on the liver of rats fed a choline-deficient (CD) diet were examined in terms of glutathione S transferase placental form (GST-P)-positive foci. Male F344 rats were given CD diet containing 0, 0.4 or 4 ppm MeIQx for 20 or 40 weeks. As controls, rats received choline-supplemented (CS) diet in the same manner. MeIQx at 4 ppm in the CD diet significantly increased both the number and area of GST-P-positive foci, the values being 2.3- and 2.1-fold at 20 weeks and 2.0- and 3.3-fold at 40 weeks, respectively, compared with those observed for CD diet alone. MeIQx at 0.4 ppm in CD diet did not affect the development of GST-P-positive foci. No influence of the heterocyclic amine was found in the CS groups, where only very small numbers of minute lesions were observed. The level of MeIQx-DNA adducts in rats given the CD diet containing 4 ppm MeIQx was 2- to 3-fold lower than that in rats given the CS diet containing 4 ppm MeIQx at 20 and 40 weeks. This result indicates that DNA adduct formation and cell proliferation are both required for the increase of GST P-positive foci in rats fed 4 ppm MeIQx in a CD diet. The above findings strongly suggest that MeIQx could be carcinogenic even at 4 ppm under CD conditions, where liver cell regeneration is continuously occurring. PMID- 8407551 TI - Expression of amphiregulin, a novel gene of the epidermal growth factor family, in human gastric carcinomas. AB - The expression of mRNA for amphiregulin (AR), a novel gene of the epidermal growth factor family, was examined in 8 human gastric carcinoma cell lines and 32 gastric carcinoma tissues as well as corresponding normal mucosa. Of the 8 gastric carcinoma cell lines, 7 expressed 1.4 kb AR mRNA at various levels. The expression of AR mRNA by TMK-1 and MKN-28 cells was increased by treatment with epidermal growth factor or transforming growth factor a. In surgical cases, all the gastric carcinoma tissues and their adjacent normal mucosa expressed AR mRNA. Interestingly, 20 (62.5%) out of 32 tumors expressed AR mRNA at higher levels than their corresponding normal mucosas (tumor/normal > or = 1.2). No obvious correlation was observed between the AR mRNA levels and the histological types or tumor staging of gastric carcinoma. Immunohistochemically, AR protein was localized to the cytoplasm and/or nucleus in tumor cells. These results suggest that AR produced by tumor cells may be involved in the pathogenesis and/or progression of human gastric carcinoma. PMID- 8407552 TI - c-Myc selectively regulates the latent period and erythroid-specific genes in murine erythroleukemia cell differentiation. AB - During the latent period of murine erythroleukemia (MEL) cell differentiation, c myc levels showed a significant change and the overexpression of the transferred c-myc gene inhibited the commitment and differentiation of MEL cells, suggesting that c-Myc may be a key molecule for the commitment. Since c-Myc may function as a DNA binding transcription factor, we examined whether c-Myc regulates the latent period genes (hsp and hsc70, MER5, Id and Spi-1 genes) and the erythroid specific genes [beta-globin, glycophorin, delta-aminolevulinic acid synthase (ALAS-E), GATA-1 and erythropoietin receptor (EpoR)] in the MEL cell transformant having transferred c-myc gene. The overexpression of c-myc gene affected the latent period genes in different ways: hsc and hsp 70 genes and Id gene were positively regulated, while expression of MER5 gene was repressed. While c-myc is thought to be involved in DNA replication, its overexpression showed no effect on the expression of proliferating cell specific nuclear antigen or DNA polymerase a. The overexpression of c-myc repressed the expression of glycophorin, ALAS-E and beta-globin genes, of the five erythroid-specific genes, but had no effect on expression of GATA-1 or EpoR gene. These results suggest that c-Myc differentially regulates the expression of the latent period and erythroid specific genes. PMID- 8407553 TI - Genetic changes and histopathological grades in human hepatocellular carcinomas. AB - Loss of heterozygosity (LOH) on chromosomes 1p, 4q, 5q, 8p, 13q, 16q, 17p, and 22q, and mutation of the p53 gene were simultaneously analyzed in 63 hepatocellular carcinomas (HCCs) with distinct histopathological grades, 80% of the tumors being from patients who had been exposed to hepatitis B virus (HBV) or hepatitis C virus (HCV). The frequencies of LOH on 8 chromosomes were 0-25% in 10 well differentiated HCCs, LOH being observed on 4q, 5q and 17p, 21-53% in 26 moderately differentiated HCCs, LOH on 8p and 17p being high, and 29-75% in 27 poorly differentiated HCCs, LOH on 17p, 4q and 8p being the most frequent. p53 gene mutation was detected in moderately and poorly differentiated HCCs at 15% and 52%, respectively, but not at all in well differentiated HCCs. Of the mutations detected, 42% were transition mutation and only 5% were CpG transition, in contrast to the high frequencies of these types of mutations in colon tumors (78% and 54%, respectively). LOH on every chromosome and p53 mutation were more frequent in more advanced tumors, and accumulation of genetic changes increased with increase of the histopathological grade. Frequency of genetic changes in HCCs from HBV-positive patients was comparable to that from HCV-positive patients. The present results suggest that accumulation of genetic changes in multiple tumor suppressor genes, especially LOH on 17p, 4q and 8p, and mutation in p53 gene, are involved in the progression of liver cancer, and LOH on 17p and 4q precedes other genetic changes. Differences in the direction of p53 mutation between HCC and colon carcinoma suggest that liver carcinogens are distinct from colon carcinogens. Furthermore, mechanisms affecting the frequency of LOH in HCCs in HBV-infected patients may be similar to those in HCV-infected patients. PMID- 8407554 TI - Infiltrating lymphocytes and accessory cells in nasopharyngeal carcinoma. AB - The infiltrating lymphocytes (LCs) and accessory cells (ACs) including dendritic cells (DCs) and monocytes/macrophages in nasopharyngeal biopsies taken from 4 groups of nasopharyngeal carcinoma (NPC) patients were observed by using an immunostaining technique and the correlation of the results to the clinical manifestations and follow-up data was examined. The findings were as follows. (1) NPCs without lymph node metastasis always had marked infiltrating LCs and DCs as compared with those with lymph node(s) metastasis. (2) Advanced NPCs with lymph node(s) involvement (T1-4N1-3M0) and a rapid development of distant metastasis followed by death within 1 year after radiotherapy always showed fewer infiltrating LCs and DCs as compared with those with lymph node(s) metastasis (T1 4N1-3M0) and having longer than 5-year survival after radiotherapy. The amount of both LCs and ACs, especially DCs, infiltrating in NPC tissues appears to be an indicator of the activity of host immune defence mechanisms against cancer and influences the progression of the neoplasm as well as the prognosis. PMID- 8407555 TI - Brefeldin A blocks the cytotoxicity of T cell receptor alpha/beta and gamma/delta cytotoxic T lymphocyte clones reacting against human autologous cancer cells. AB - We studied the effector mechanism of T cell receptor (TCR) alpha/beta- and gamma/delta-type cytotoxic T lymphocyte (CTL) clones that react with human autologous tumor cells. Treatment of tumor cells with a fungal antibacterial reagent, brefeldin A (BFA), resulted in the inhibition of cytotoxicity of an autologous tumor (HST-2)-specific CD8+ TCR alpha/beta-type CTL, TcHST-2. Other anti-metabolites such as chloroquine, cycloheximide and colchicine did not affect the cytotoxicity. The cell-surface antigen expression, including MHC class I molecules, was not influenced by BFA treatment. Furthermore, BFA did not influence the cytotoxicity of lymphokine-activated killer cells and natural killer cells. Since BFA blocks the transport of peptides from endoplasmic reticulum to the Golgi apparatus, the above data suggest that BFA could affect washing out of the peptide fragments from the MHC class I groove. Consequently, target tumor cells were protected from killing by CTL. Moreover, we obtained a CD4-, 8-, TCR gamma/delta-type (V delta 1+) CTL clone, TcHOT, that reacts against an autologous ovarial carcinoma, HOT. BFA could also inhibit this cytotoxicity, and it is likely that different presenting molecules other than MHC class I proteins participate in the cytotoxicity of this TCR gamma/delta-type CTL. These studies suggest that both TCR alpha/beta- and gamma/delta-type CTL may require antigenic peptides that are most likely derived from the BFA-sensitive, intracellular endogenous target proteins. PMID- 8407556 TI - Enhanced tumor localization of radiolabeled Fab fragments of monoclonal antibody A7 in nude mice bearing human pancreatic carcinoma xenografts. AB - Much recent research has been directed toward the use of monoclonal antibodies (MAb) for the immunodetection of solid tumors. In pancreatic cancer, the results of conventional immunoscintigraphy using intact MAb remain disappointing. Clear immunoscintigraphy with radiolabeled MAb requires a high tumor tissue/blood ratio of radioactivity and a low normal tissue/blood ratio of radioactivity. In this study, 125I-labeled Fab fragments produced by papain digestion of MAb A7 were injected intravenously into nude mice bearing a human pancreatic cancer (HPC-YS) xenograft previously shown to react specifically with MAb A7. The radioactivity of tumors and normal organs was subsequently measured. The tumor tissue/blood ratio of 125I-labeled Fab fragments of MAb A7 was 1.00 +/- 0.24 and 9.68 +/- 2.54 at 2 and 24 h after injection, respectively. The tumor tissue/blood ratio of radioactivity was significantly higher than those of normal organs at 24 h after injection. Moreover, the tumor tissue/blood ratio of 125I-labeled Fab fragments of MAb A7 was greater than that of intact MAb A7, although the 125I-labeled Fab accumulation level was much less than that of 125I-labeled intact MAb A7 in the tumor. When mice bearing tumors which did not react with MAb A7 were studied, 125I-labeled Fab fragments did not specifically localize to the tumors. These results suggest that Fab fragments of MAb A7 may be suitable carriers of radionuclides for the immunodetection of human pancreatic cancer. PMID- 8407557 TI - Abnormal biantennary sugar chains in human choriocarcinoma. PMID- 8407558 TI - Guidelines for analysis and reporting of clinical trials in oncology. AB - When analyzing and reporting the results of clinical trials, investigators should follow a simple approach. The purpose of a trial is to estimate an effect or treatment difference, which if present would have clinical utility when treating new patients. Procedures or methods that do not facilitate precisely and impartially estimating and reporting the treatment effect are likely to mislead investigators. Most often in clinical trials, investigators are interested in estimates of risk ratios (specifically odds or hazard ratios) between the treatment groups or levels of a prognostic factor. These simple ideas suggest that the most useful results from clinical trials will be estimated risk ratios and their confidence limits. Especially in cancer, where disease progression, recurrence, and death are common events following treatment, estimates of risk difference are very relevant. Hypothesis tests and associated P-values, although often (or exclusively) reported, are of lesser utility because they do not fully summarize the data. These recommendations may be seen by some investigators to be contrary to accepted practice. It is true that they are somewhat contrary to common practice but their general acceptance is evident in many journals and presentations by clinical trial methodologists. Despite some disagreement among statisticians regarding the need for adjustment of analyses for imbalanced prognostic factors, it is helpful to see if treatment effects change after accounting for imbalances. When this occurs, it may be of clinical interest. Although we discourage analyses that exclude any patients who meet the eligibility criteria, some circumstances will require that this be done (e.g., when a patient refuses to participate after randomization). Investigators should report, and emphasize as primary, those analyses that include all eligible patients. It is our hope and belief that analysis and reporting of trial results along the guidelines suggested here will result in impartial and useful information for journal readers. PMID- 8407559 TI - Benign breast disease as a breast cancer risk in Japanese women. AB - A hospital-based retrospective cohort study of benign breast disease (BBD) as a risk factor of future breast cancer (BC) development was conducted. Four hundred and twenty-eight patients with biopsied BBD were followed-up for a median period of 8 years, together with age-matched women with normal breasts (normal control) and BC patients (cancer control), at the ratio of 1:2:2. Twenty-one breast cancers developed, 7 in the cases, 4 in the normal controls, and 10 in the BC controls, showing the relative risk (RR) with 95% confidence intervals (CI) to be 3.5 (1.03-11.9) in the cases with respect to the normal controls. The RR of the cases is not lower than that of contralateral breast cancer incidence. There were no significant differences in the risks of cancers in other organs among the groups. Pathological examination revealed that only atypical hyperplasia increased the RR of BC, as compared with the normal control breast group, or with non-proliferative disease. These results suggest that in a low-risk country, Japan, BBD is a definite risk factor for BC development as in high-risk countries. PMID- 8407560 TI - Enhancing effects of calcium-deficient diet on gastric carcinogenesis by N-methyl N'-nitro-N-nitrosoguanidine in Wistar rats. AB - The effects of ad libitum feeding of calcium-deficient diet on the incidence, number and histological types of gastric cancers induced by N-methyl-N'-nitro-N nitrosoguanidine (MNNG) were investigated in male Wistar rats. Rats were fed standard pellet diet containing 0.5% (normal-calcium diet) or 0.01% calcium (calcium-deficient diet) after oral treatment with MNNG for 25 weeks. Oral administration of the calcium-deficient diet resulted in a significant increase in the incidence, but not the number, of gastric cancers in experimental Week 52. However, it did not affect the histological types of cancer. The calcium deficient diet also caused a significant increase in tissue norepinephrine concentration of the antral portion of the gastric wall and in the labeling index of the antral epithelial cells. These findings indicate that the calcium deficient diet enhanced gastric carcinogenesis and suggest that its effect may be related to increase in norepinephrine in the gastric wall and consequent stimulation of proliferation of antral epithelial cells. PMID- 8407561 TI - Enhancing effects of estrogens on endometrial carcinogenesis initiated by N methyl-N-nitrosourea in ICR mice. AB - The present study was undertaken to examine the effects of estrogens, such as estrone (E1), 17 beta-estradiol (E2) and estriol (E3), on endometrial carcinogenesis initiated by N-methyl-N-nitrosourea (MNU) in mice. A total of 120 female ICR mice received MNU solution (1 mg/100 g body wt.) and normal saline at 10 weeks of age into their left and right uterine corpora, respectively. One week later, they were divided into four groups and treated as follows: Group 1 (30 mice) was given 25 ppm E1-containing diet; and Group 2 (30 mice) was fed 5 ppm E2 containing diet; Group 3 (30 mice) was given 25 ppm E3-containing diet; and Group 4 (30 mice) was fed the basal diet alone. At the termination of the experiment (Week 30), all surviving animals were autopsied and histopathological examinations revealed that endometrial adenocarcinomas had developed in all groups. The incidence of adenocarcinomas in the MNU-treated uterine corpus in Group 1 (25 ppm E1-feeding, 9/23, 39%) was significantly higher than that in Group 4 (basal diet, 3/26, 12%, P < 0.05). Also, the incidences of adenocarcinomas in the MNU-treated uterine corpus in Groups 2 (5 ppm E2-feeding, 8/24, 33%) and 3 (25 ppm E3-feeding, 7/26, 28%) were higher than in Group 4, but the difference was not statistically significant. Feeding of diet containing E1, E2 and E3 increased the incidences of the preneoplastic endometrial lesions (atypical, adenomatous or cystic glandular hyperplasia). In the uterine cervix, small numbers of squamous cell carcinomas, dysplasias or hyperplasias were occasionally found in all groups. These results indicate enhancing effects of the above three types of estrogens on the endometrial carcinogenesis induced by MNU in ICR mice. PMID- 8407562 TI - Comparison of K-ras oncogene activation in pancreatic duct carcinomas and cholangiocarcinomas induced in hamsters by N-nitrosobis(2-hydroxypropyl)amine. AB - The presence of K-ras point mutations in pancreatic duct carcinomas and cholangiocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine (BHP) in Syrian hamsters was investigated by single-strand conformation polymorphism analysis of polymerase chain reaction products from frozen fresh materials in order to clarify the K-ras mutation rates in those two carcinomas induced simultaneously by one carcinogen, BHP. In the examined pancreatic duct carcinomas, 10 out of 16 were positive for a mutation in codon 12 while 3 out of 12 cholangiocarcinomas demonstrated mutation of K-ras gene. G-to-A transition was detected in the second position of codon 12 in both pancreatic carcinomas and cholangiocarcinomas. These results suggest that the role of genetic alteration in carcinogenesis may differ with the target organ, even when initiation is with the same carcinogen. PMID- 8407563 TI - Identification of K-ras oncogene mutations in the pure pancreatic juice of patients with ductal pancreatic cancers. AB - Pancreatic cancer is detected on the basis of morphological changes delineated by means of various image-diagnostic methods. However, differentiation between chronic pancreatitis and pancreatic cancer, especially at the early stage, is not always simple when based upon the morphological changes alone. Therefore, we attempted to elucidate K-ras mutations in the sediment of pure pancreatic juice (PPJ) containing exfoliated ductal pancreatic cancer cells. PPJ was collected endoscopically from 20 patients with pancreatic cancer (PC) and 18 patients with chronic pancreatitis (CP). Polymerase chain reaction and allele specific oligonucleotide dot blot hybridization for K-ras mutations were performed with the DNA extracted from these samples. A K-ras mutation at codon 12 was identified in the PPJ of 11/20 (55%) of the patients with PC. On the other hand, the same mutation was not identified in the PPJ of any patient with CP. Moreover, K-ras mutations at codons 13 and 61 were not recognized in the PPJ of any patient with either PC or CP. These findings suggested that the presence of a K-ras mutation at codon 12 in PPJ would be useful in confirming the diagnosis of PC. PMID- 8407564 TI - High sensitivity of LEC rats with chronic hepatitis to hepatocarcinogenesis: decreases in unscheduled and replicative DNA synthesis of the hepatocytes. AB - We carried out the following three experiments to clarify the mechanism of hepatocarcinogenesis in Long-Evans Cinnamon (LEC) rats. (1) Sensitivity to diethylnitrosamine (DEN): LEC rats (8 and 25 weeks old) without and with hepatitis and age-matched F344 rats were administered an intraperitoneal injection of a low dose of DEN. Eight weeks after the injection, the numbers of glutathione-S-transferase placental-form (GST-P)-positive foci in the 33-week-old LEC rat liver were significantly higher than those in the livers of the other three groups of rats. (2) Potential for unscheduled DNA synthesis (UDS): Isolated hepatocytes of 25-week-old LEC rats with chronic hepatitis showed about one-third the level of UDS induced by UV irradiation, as compared to that of age-matched F344 rats, while no significant difference was found between the UDS of isolated hepatocytes of 8-week-old LEC rats and age-matched F344 rats. (3) Potential for proliferation: Isolated hepatocytes from 8-week-old LEC rats responded well to epidermal growth factor (EGF) in culture, to almost the same degree as F344 rat hepatocytes, while a remarkable decrease in the responsiveness of hepatocytes isolated from 25-week-old LEC rats to EGF was found. These results suggested that LEC rat hepatocellular carcinoma could be naturally initiated after the onset of hepatitis by carcinogens contaminating food and the environment, probably due to the reduction of DNA repair activity, after which initiated hepatocytes selectively proliferate in response to growth stimuli endogenously produced as a result of continuous loss of hepatocytes (chronic hepatitis), because of a decrease in growth activity of non-initiated hepatocytes. PMID- 8407565 TI - Immunohistochemical analysis of cathepsin B expression in human lung adenocarcinoma: the role in cancer progression. AB - Production of cathepsin B by tumor cells has been linked to metastatic potential in several experimental models. Sections of 95 primary lung adenocarcinomas were examined for expression of cathepsin B using a standard avidin-biotin immunohistochemical technique. Staining for cathepsin B was observed in 22.1% of all cases and 28.0% of those of the Clara cell type. In Clara cell adenocarcinomas, cathepsin B expression correlated with positive lymph node status, presence of distant metastases, and poor prognosis (P < 0.05). However, no correlation with clinical outcome was observed in other cell types. Our data suggest that cathepsin B may be involved in invasion and metastasis in Clara cell lung adenocarcinoma. PMID- 8407566 TI - Follow-up study of patients treated with monoclonal antibody-drug conjugate: report of 77 cases with colorectal cancer. AB - A total of 77 patients with advanced colorectal cancer, including postoperative patients with liver, lung and peritoneal metastases, were treated with single or multiple injections of monoclonal antibody A7-neocarzinostatin (A7-NCS). A follow up study of the patients treated with A7-NCS was done and the clinical outcome was compared with that of patients given other chemotherapies. In the postoperative patients with liver metastasis, the A7-NCS treatment prolonged survival time when compared with systemic administration of anticancer drugs, while it showed a similar survival time to chemoembolization using multiple anticancer agents suspended in a lipid contrast medium. Among the patients who underwent surgical resection of primary cancer, with or without liver metastasis, there was no difference in overall 5-year survival rate between the group treated with A7-NCS and the group treated with the other chemotherapies. However, the survival time of the patients treated with A7-NCS was longer than that of the patients treated with the other chemotherapies. In addition, the patients given a higher dose of A7-NCS had a longer survival time than the patients given a lower dose of A7-NCS. Human anti-mouse antibody was detected in all the A7-NCS-treated patients examined. There were no serious side effects in any of the patients given A7-NCS. Thus, this study indicates that the A7-NCS treatment is safe and useful for colorectal cancer patients, though some problems remain, such as optimization of injection dose, route, interval, etc., and overcoming human anti mouse antibody development. PMID- 8407567 TI - Detailed characterization of a high-molecular-weight glycoprotein secreted by lung cancer cells. AB - A cancer-associated, high-molecular-weight glycoprotein antigen (6B3.Ag) recognized by monoclonal antibody 6B3 was purified from culture medium of human large cell lung carcinoma cell line (HLC-2) and characterized biochemically and immunochemically. The 6B3.Ag was purified more than 1,200-fold with a yield of 30% by salting out, precipitation by acidification at pH 4.5, and chromatographies on Sepharose 4B and concanavalin A-Sepharose. The molecular weight of 6B3.Ag is approximately 1,000,000 and the molecule is a homodecamer of 94,000 subunits. The 6B3.Ag is a glycoprotein containing 22.9% sugars, consisting of both N- and O-glycoside chains. The N-terminal 19 amino acids were determined and only 4 out of 19 amino acid residues were different from those of an antigen, L3, secreted by lung carcinoma cell line Calu-1. The serum level of 6B3.Ag was determined in normal adults as well as patients with various diseases by enzyme linked immunosorbent assay. The mean serum level of 6B3.Ag was 3.1 micrograms/ml, ranging from 1.6 to 6.2 micrograms/ml in 131 healthy adults. When the cut-off value was set at 6.2 micrograms/ml, the incidence of positive values in the sera was elevated not only in malignant diseases such as hepatoma (73%) and leukemia (62%), but also in benign diseases such as chronic hepatitis (42%) and liver cirrhosis (63%). While the incidence of positive values was elevated in advanced liver diseases, namely, chronic hepatitis, liver cirrhosis and hepatoma, the cancer specificity of 6B3.Ag did not appear to be high. PMID- 8407568 TI - Aberrant alpha 1-->2fucosyltransferases found in human colorectal carcinoma involved in the accumulation of Leb and Y antigens in colorectal tumors. AB - Evidence indicates that the presence of aberrant alpha 1-->2fucosylation pathways is responsible for the accumulation of large quantities of Le(b) and Y antigens in human colorectal carcinoma. Significantly higher activities of alpha 1-->2 as well as alpha 1-->3 and alpha 1-->4fucosyltransferases were found in most of the tissues from carcinoma than in the adjacent normal tissues and in healthy subjects. alpha 1-->2Fucosyltransferases associated with the synthesis of Le(b) (Fuc alpha 1-->2Gal beta 1-->3[Fuc alpha 1-->4]GlcNAc beta) and Y (Fuc alpha 1- >2Gal beta 1-->4[Fuc alpha 1-->3]GlcNAc beta) structures from Le(a) (Gal beta 1- >3[Fuc alpha 1-->4]GlcNAc beta) and X (Gal beta 1-->4[Fuc alpha 1-->3]GlcNAc beta) ones, respectively, were demonstrated in colorectal carcinomas and in colorectal carcinoma cell lines (COLO201, LS174T and SW1116). The activation of alpha 1-->2fucosyltransferase with such new substrate specificities in colorectal carcinoma might result in the preferential synthesis of Le(b) and Y structures from Le(a) and X rather than from H type 1 and H type 2 structures. PMID- 8407569 TI - Immunohistochemical localization of insulin-like growth factor I (IGF-I) in the rat epididymis. AB - The distribution of insulin-like growth factor I (IGF-I) was studied by immunohistochemistry during postnatal development of the rat epididymis. At 2 weeks the immunoreactivity was mainly located along the cytoplasmic apical border in both the caput and the cauda epididymidis. A slight immunolabeling was present in the myofibroblastic cells. Afterward, the epithelial immunoreactivity was minimal at 4 weeks and increased progressively after the 6th week, especially in the apical and subapical cytoplasmic compartments of the caput epididymidis. The labeling of the epithelial cells of the cauda epididymidis was restricted to the apical cytoplasmic area. Immunolabeling was also found in the myofibroblastic cells and was more intense after 6 weeks. The variations of the pattern of distribution support the hypothesis of a physiological role for IGF-I in the regulation of epididymal functions. PMID- 8407570 TI - Recovery of pituitary-gonadal function in male rats after long-term suppression induced by a single injection of microcapsules of LH-RH antagonist cetrorelix (SB 75). AB - The clinical utility of luteinizing hormone-releasing hormone (LH-RH) analogs can be greatly enhanced by a sustained delivery system, which could maintain elevated peptide levels in the blood for prolonged periods of time, up to several weeks. Recently, we developed long-acting microcapsules and microgranules of the LH-RH antagonist SB-75. In this study, we examined the suppressive effects of a single injection of microcapsules of antagonist SB-75 on gonadotropin and testosterone secretion, as well as on fertility, in male rats and the reversibility of those effects. Serum SB-75 levels were measured by RIA. A dose of 20 mg of microcapsules/rat containing 3.58 mg of antagonist in poly(D,L-lactide-co glycolide), administered intramuscularly produced SB-75 levels higher than 20 ng/ml for approximately 24 days, and a significant elevation was maintained until day 90. Serum testosterone was decreased to castration values for 164 days and LH levels were suppressed below the detection limit of the RIA for a period of 102 days. Serum FSH was suppressed by more than 90%, as compared to control animals, for a period of 58 days and remained significantly decreased until day 164 after the injection. This treatment also caused a significant decrease in the weights of the testes, seminal vesicles, and ventral prostate 30 days after peptide administration. The histology of the testes from the treated rats showed that spermatogenesis was totally depressed. No mature elongated or round spermatids were found in the seminiferous tubules, spermatocytes being the most advanced germ cell form in 99.5% of the testicular tubules.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407571 TI - Pituitary adenylate cyclase activating polypeptide (PACAP) can cross the vascular component of the blood-testis barrier in the mouse. AB - Pituitary adenylate cyclase activating polypeptide (PACAP), present in highest concentrations in the hypothalamus and testes, affects the release of LH, FSH, and prolactin, as well as Sertoli cell function. We examined the ability of the 38-amino acid form of PACAP labeled with 125I (I-P38) to cross the vascular component of the blood-testis barrier. The unidirectional influx constant (Ki) was 4.23 x 10(-3) ml/g-minute, which is about 5 times faster than the entry of LH and about 17 times faster than that of serum albumin. Entry occurred in part by a saturable transport system, with 20 micrograms/mouse of unlabeled P38 inhibiting transport by 40%. An analog of peptide T, which like PACAP is related to vasoactive intestinal polypeptide and has been found to have its own saturable transport system into the brain, did not alter the uptake of I-P38 by the testes. A dose of 10 micrograms/mouse, but not of 20 micrograms/mouse, was associated with a contraction of the vascular space of the testes. HPLC confirmed that a small but persistent percentage of the radioactivity recovered from the testes represented intact I-P38. These results suggest that circulating P38 may contribute to the testicular pool of PACAP, which may play an active role in the function of the testes. PMID- 8407572 TI - The effects of chronic administration of pyrimethamine on spermatogenesis and fertility in male rats. AB - The present study examines whether the antifertility effects of pyrimethamine (PYR), an inhibitor of dihydrofolate reductase, are mediated by a reduction in intratesticular testosterone (T) concentrations or whether PYR exerts its effect by a cytotoxic insult to spermatogenic cells that is independent of intratesticular testosterone. Adult male rats were treated daily with 100 mg/kg (n = 16) or 400 mg/kg (n = 16) of PYR in honey for 8 weeks. Control rats (n = 16) received honey without PYR. Eight weeks after treatment, five rats from each PYR treated group and five control rats were mated with normal cycling female rats, and fertility was assessed. These rats were euthanized after the fertility trial; testis weight, testicular sperm, and epididymal sperm counts were determined, and serum levels of T, LH, FSH, and seminiferous tubule fluid T (STF-T) concentrations were measured by RIA. Testes from three rats per group were perfusion-fixed for histological evaluation. PYR was discontinued in the remaining rats for 8 weeks and similar parameters were evaluated after 8 weeks of recovery. PYR (100 mg/kg/day) treatment for 8 weeks did not have any effects on organ weights, testicular and epididymal sperm counts, and hormone levels when compared to controls. In contrast, PYR (400 mg/kg/day) treatment significantly reduced testis and epididymis weights, testicular and epididymal sperm counts, and fertility. Despite these effects, serum T, LH, FSH, and STF-T concentrations were not altered.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407573 TI - Testicular intravascular volume and microvessel mitotic activity: effect of experimental varicocele. AB - Testicular blood flow increases bilaterally in animals with experimental left varicocele (ELV). The present experiments were undertaken to determine if increases in testicular intravascular volume and capillary growth were potential contributors to this increase in flow. Testicular intravascular volume (microliters/g) was estimated by determining testicular vascular 125I-albumin space. Microvessel mitotic activity was estimated by examining 3H-thymidine incorporation into endothelial nuclei, as assayed by silver grain localization over microvessel endothelial nuclei after exposure of testes to 3H-thymidine in vivo. Testicular intravascular volume was 25-30 microliters/g testis in control testes and in ELV animals was increased an average of 9% and 28% in right and left testes, respectively. This increase was statistically significant (P < 0.05) on the left side only. Approximately 1.5-2.0% of testicular microvessel endothelial nuclei exhibited silver grain localization, and ELV was not associated with a change in these values in either right or left testis. Ancillarily, there was a modest but significant association between microvessel mitotic activity and mitotic activity in adjacent seminiferous tubules. It was concluded that bilateral increases in testicular blood flow after ELV are not uniquely the result of bilateral expansion of intravascular volume and growth of the testicular capillaries. It is speculated, however, that mitogenic factors from the seminiferous tubules potentially affect mitotic activity of the adjacent microvasculature. PMID- 8407574 TI - Soluble Fc gamma RIII (CD16) and immunoglobulin G levels in seminal plasma of men with immunological infertility. AB - Receptors for the Fc region of the immunoglobulin G (IgG) (Fc gamma R) have been recognized as a link between humoral and cellular immune responses. A soluble form of Fc gamma RIII (CD16) has been found in seminal plasma (SP), which may modulate immunosuppression of antisperm immune responses in the male and female reproductive tracts. SP from some individuals apparently have lower levels of Fc gamma RIII, but it is not known whether the diminished activities are due to low receptor concentration or steric interference from IgG. To test the hypothesis that different levels are due to steric interference, relative levels of Fc gamma RIII were measured in SP using monoclonal antibody 3G8 in an amplified enzyme linked immunosorbent assay (ELISA) system. Men who were positive for antisperm antibodies (ASA) by Sperm Mar and direct immunobead assay (N = 26) and negative for ASA (N = 26) were tested. Individuals who were ASA positive had lower detectable levels than those who were ASA negative (t = 1.99, P = 0.05). Therefore, variation in Fc gamma RIII levels may be due to steric interference from IgG. IgG subclass concentrations in SP of both groups were determined using an ELISA method and compared to Fc gamma RIII levels. Slight correlations were seen for IgG1 (r2 = 0.237, P < 0.001), IgG2 (r2 = 0.204, P < 0.001), and total IgG (r2 = 0.299, P < 0.001) in relation to Fc gamma RIII levels in ASA-negative SP specimens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407575 TI - Motile sperm in human testis biopsy specimens. AB - We prospectively studied 62 consecutive infertile men who underwent 100 intraoperative wet prep cytological examinations of testis biopsy material obtained simultaneously with permanently fixed specimens. Wet preps were performed by placing a small sample of fresh testicular tissue on a slide, adding a drop of Ringer's lactate, and compressing the specimen under a glass coverslip. Among these 100 wet preps, complete sperm with tails were identified in 62 specimens, of which 44 contained nonmotile sperm and 18 contained motile sperm. Reproductive tract obstruction was documented in 65 testes (65%) on subsequent reconstructive surgery and/or inferred from histological evaluation, including mean mature spermatid counts on the permanent sections fixed in Bouin's solution. Obstruction was absent in the remaining testes (35%). All 18 testes with motile sperm found on wet prep were obstructed. These testes were also found to have complete spermatogenesis, a category selected to include normal spermatogenesis and slight hypospermatogenesis, determined by examination of the permanently fixed sections. The finding of motile vs. nonmotile sperm on a wet prep has positive predictive values of 100% vs. 81% for the presence of reproductive tract obstruction and 94% vs. 86% for complete spermatogenesis, respectively. The presence of motile sperm in human testis biopsy specimens is a novel finding. When any complete sperm with tail is found in a testis biopsy wet prep, obstruction is likely. When motile sperm are present, obstruction is almost certain, and immediate exploration and reconstructive surgery can be justified.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407576 TI - Evidence that membrane stress contributes more than lipid peroxidation to sublethal cryodamage in cryopreserved human sperm: glycerol and other polyols as sole cryoprotectant. AB - One effect of cryopreservation on human sperm is sublethal cryodamage, in which cell viability post-thaw is lost more rapidly at later times than in fresh cells. We hypothesized two modes of sublethal cryodamage: one is peroxidation-related involving plasma membrane damage due to lipid peroxidation; the other is membrane stress-related involving membrane embrittlement during phase transitions occurring during freeze-thaw. If the peroxidation-related mode contributed substantially to sublethal cryodamage, the hypothesis predicts that lipid peroxidation inhibitors should reduce this damage. To test this prediction, we examined the effect of the lipid peroxidation inhibitors, hypotaurine, bovine serum albumin (BSA), and alpha-tocopherol (Vit. E) on the time to loss of motility (TLM), taken as a measure of cell viability over time, for sperm samples cryopreserved in glycerol plus egg yolk medium. These agents had no effect on TLM of these samples, indicating that this mode contributes little to sublethal cryodamage. If the membrane stress-related mode contributed, the hypothesis predicts rapid recovery of motility in the presence of egg yolk plus glycerol, but slow recovery in the presence of glycerol alone. It also predicts that an appropriate polyol may be both necessary and sufficient for cryopreservation. In the presence of egg yolk plus glycerol, motility recovery was complete within 5 minutes, but the percent motile cells then decreased linearly with time. With glycerol alone in the range 3-12%, at 5 minutes post-thaw the percent motile cells was 5-10%, but by 40 minutes post-thaw had risen to 60-80%, approaching that in the fresh sample, and was maintained up to 4 hours. In the absence of glycerol, the percentage of motile cells post-thaw was nil and remained nil up to 4 hours. The polyols, erythritol, ribitol, and sorbitol had similar effects to that of glycerol, but the recovery of motility was not as complete. These results indicate that the membrane stress-related mode contributes substantially to sublethal cryodamage. They also indicate that glycerol and other polyols can function alone as cryoprotectants, but that recovery of motility is slow in these systems. PMID- 8407577 TI - Sampling factors influencing accuracy of sperm kinematic analysis. AB - Sampling conditions that influence the accuracy of experimental measurement of sperm head kinematics were studied by computer simulation methods. Several archetypal sperm trajectories were studied. First, mathematical models of typical flagellar beats were input to hydrodynamic equations of sperm motion. The instantaneous swimming velocities of such sperm were computed over sequences of flagellar beat cycles, from which the resulting trajectories were determined. In a second, idealized approach, direct mathematical models of trajectories were utilized, based upon similarities to the previous hydrodynamic constructs. In general, it was found that analyses of sampling factors produced similar results for the hydrodynamic and idealized trajectories. A number of experimental sampling factors were studied, including the number of sperm head positions measured per flagellar beat, and the time interval over which these measurements are taken. It was found that when one flagellar beat is sampled, values of amplitude of lateral head displacement (ALH) and linearity (LIN) approached their actual values when five or more sample points per beat were taken. Mean angular displacement (MAD) values, however, remained sensitive to sampling rate even when large sampling rates were used. Values of MAD were also much more sensitive to the initial starting point of the sampling procedure than were ALH or LIN. On the basis of these analyses of measurement accuracy for individual sperm, simulations were then performed of cumulative effects when studying entire populations of motile cells. It was found that substantial (double digit) errors occurred in the mean values of curvilinear velocity (VCL), LIN, and MAD under the conditions of 30 video frames per second and 0.5 seconds of analysis time. Increasing the analysis interval to 1 second did not appreciably improve the results. However, increasing the analysis rate to 60 frames per second significantly reduced the errors. These findings thus suggest that computer-aided sperm analysis (CASA) application at 60 frames per second will significantly improve the accuracy of kinematic analysis in most applications to human and other mammalian sperm. PMID- 8407578 TI - Performance and comparison of CASA systems equipped with different phase-contrast optics. AB - Most computer-aided sperm analysis (CASA) systems for human spermatozoa use positive phase-contrast optics. In this study the performance of one of these (HTM) and two other systems using negative phase optics (IVOS and SM) were assessed by scrutinizing the video-playback of the analyzed images for errors in the recognition of motile and immotile spermatozoa and in the tracking of motile cells. Whereas both the HTM and IVOS provided accurate measurements of motile sperm concentrations, errors in total sperm concentrations and motility percentages were greater in the IVOS than the HTM. However, IVOS appeared to be more accurate in tracking motile spermatozoa. Individual sperm track data suggested that velocity measurements may be biased by the elimination of incomplete tracks from the analysis. With the SM system, discrimination of immotile spermatozoa from non-sperm particles in semen was poor despite an additional algorithm for tail detection. The use of negative phase optics in CASA may be superior to that of positive phase in the analysis of motile spermatozoa but inferior in the recognition of immotile spermatozoa. PMID- 8407579 TI - Structure-activity relationships of vancomycin-type glycopeptide antibiotics. PMID- 8407580 TI - AS-183, a novel inhibitor of acyl-CoA: cholesterol acyltransferase produced by Scedosporium sp. SPC-15549. AB - A novel compound, AS-183, which inhibits acyl-CoA: cholesterol acyltransferase (ACAT), was isolated from the culture broth of a fungus, Scedosporium sp. SPC 15549. AS-183 inhibited ACAT activity in an enzyme assay system using rabbit liver microsomes with an IC50 value of 0.94 microM. AS-183 also inhibited cholesterol ester formation in HepG2, CaCo2, and THP-1 cells with IC50 values of 18.1, 25.5, and 34.5 microM, respectively. PMID- 8407581 TI - Mureidomycins E and F, minor components of mureidomycins. AB - Mureidomycins (MRDs) E and F were isolated from a culture filtrate of Streptomyces flavidovirens SANK 60486 which produces MRDs A approximately D. They possessed the same molecular formulae, C39H48N8O12S and very similar UV, IR and NMR spectra, but differed clearly from each other in HPLC profile. From the hydrolysates of MRDs E and F, 8-hydroxy-1,2,3,4-tetrahydro-3-isoquinoline carboxylic acid and 6-hydroxy-1,2,3,4-tetrahydro-3-isoquinoline carboxylic acid were detected, respectively, which were not detected from those of MRDs A approximately D. They showed strong anti-pseudomonal activity but less active than MRD A. MRDs E and F were synthesized from MRD A and formaldehyde through Pictet-Spengler reaction. PMID- 8407582 TI - Phthoxazolin A, a specific inhibitor of cellulose biosynthesis from microbial origin. I. Discovery, taxonomy of producing microorganism, fermentation, and biological activity. AB - A new inhibitor of cellulose biosynthesis named phthoxazolin A was discovered as a metabolite of Streptomyces sp. OM-5714. A newly established screening method, which utilized a cellulose-containing fungus, Phytophthora sp. as a test organism, was successfully employed for discovery of the compound. Phthoxazolin A, C16H22N2O3 (MW of 290), is a lipophilic triene compound with an oxazole moiety. It has moderate antifungal activity against Phytophthora spp. only and has potent herbicidal activity. Phosphate-limited fermentation conditions favored production of the active compound. PMID- 8407583 TI - Phthoxazolin A, a specific inhibitor of cellulose biosynthesis from microbial origin. II. Isolation, physico-chemical properties, and structural elucidation. AB - Phthoxazolin A is a new inhibitor of cellulose biosynthesis produces by Streptomyces sp. OM-5714. The active compound was isolated, and the structure was elucidated by spectrometric analyses. PMID- 8407584 TI - Microbial glycosidation of some anthracycline antibiotics by an antibiotic negative mutant of aclarubicin producer. AB - Microbial conversion of anthracyclinone monosaccharides using aclarubicin negative mutant of Streptomyces galilaeus was found to produce anthracyclinone disaccharides which had either rhodinose or 2-deoxyfucose as an additional sugar. By this conversion we obtained twelve new anthracyclines from seven anthracyclines which had rhodosamine, N-monomethyldaunosamine or daunosamine at C 7 as a glycosidic sugar. All products had a reduced cytotoxic activity in comparison with those of parent compounds. However, some of them showed a therapeutically improved antitumor effects against L1210 leukemia in vivo. PMID- 8407585 TI - Studies on cell growth stimulating substances of low molecular weight. Part 2. Exfoliazone and lavanducyanin, potent growth promoting substances of rat liver cell line, RLN-8, produced by Streptomyces exfoliatus and Streptomyces aeriouvifer. AB - Exfoliazone and lavanducyanin isolated from Streptomyces exfoliatus BT-38 and Streptomyces aeriouvifer CL-190, respectively, showed strong growth promoting activities to liver cell RLN-8 established from normal Donryu rat. When RLN-8 cells were cultured in Eagle's minimal essential medium containing 1% fetal bovine serum, exfoliazone significantly stimulated the growth of RLN-8 cells. However, no effect was observed under serum-free conditions. Effective dose of exfoliazone was at the range of 0.004-0.1 microgram/ml. Cell proliferation was confirmed by MTT assay and by the increases of cell number and DNA synthesis. Lavanducyanin also stimulated the growth of RLN-8 cells in the same medium. It showed growth promoting activity at lower concentrations than exfoliazone and the effective dose was at the range of 0.0001-0.06 microgram/ml. Analogous compounds of exfoliazone and lavanducyanin also promoted the growth of RLN-8 cells. In addition, exfoliazone and lavanducyanin enhanced the growth of NIH 3T3 and T601 cells. These results indicate that exfoliazone, lavanducyanin and their related compounds seem to be a new type of growth promoting substances with low molecular weight produced by microorganisms, and that they can partially substitute for functions of serum. Since 12-O-tetradecanoylphorbol-13-acetate (TPA) did not show the growth promoting activities under the same conditions, the action mechanism(s) of exfoliazone and lavanducyanin are different from that of TPA. PMID- 8407586 TI - 9a,11-cyclic carbamates of 15-membered azalides. AB - The novel 9a,11-cyclic carbamates (13-15) of 9-deoxo-9a-aza-9a-homoerythromycin A (4) have been prepared and characterized by 1H-1H and 1H-13C 2D NMR spectroscopy. When compared to azithromycin (1) or its 6-O-methyl derivative (2), the new bicyclic 15-membered azalides exhibited substantially decreased antibacterial activities in vitro. PMID- 8407587 TI - Antifungal activities of pradimicin derivatives modified at C4'-amino group. AB - In order to explore potent derivatives of pradimicins (PRMs), modification of their C4'-amino group was carried out. 4'-N-Cyano (1,2), 4'-deamino-4' nitroguanidino (4), 4'-deamino-4'-ureido (7-9) and 4'-deamino-4'-thioureido (10) derivatives were synthesized by trimethylsilylation of PRMs A and C, followed by condensation with appropriate reagents. 4'-Deamino-4'-guanidino (5) and 4' deamino-4'-amidino (6) derivatives were synthesized by catalytic hydrogenation of 4 and 2, respectively. 4'-N-Nitroso derivative 3 was prepared by treatment of PRM A with nitrous acid. Among these compounds, the 4'-N-cyano derivative of PRM C (2) exhibited in vitro and in vivo antifungal activities comparable to the parent compounds together with good water-solubility. PMID- 8407588 TI - Synthesis and anthelmintic activity of 13-alkoxymilbemycin derivatives. AB - The synthesis of milbemycin derivatives having alkyloxy groups at the C-13 position was studied and a series of these analogs of milbemycin A4 and A3 was prepared by the reaction of 13-iodomilbemycin with a variety of alcohols. 13 beta Phenethyloxy derivatives were found to possess excellent anthelmintic activity. Especially, the activities of derivatives with N-substituted 4-aminophenethyloxy groups were comparable to or superior to ivermectin against Nippostrongylus brasiliensis in rats. PMID- 8407589 TI - Synthesis of 3-deoxy-3,4-didehydro derivatives of 5-O-mycaminosyltylonolide, 5-O (4-deoxymycaminosyl) tylonolide, and desmycosin. AB - The 3-deoxy-3,4-didehydro derivatives of 5-O-mycaminosyltylonolide, 5-O-(4 deoxymycaminosyl)tylonolide, and desmycosin have been prepared by treatment of the corresponding 3-O-sulfonyl derivatives with NaI in 2-butanone as the key step. The mechanistic difference in the formation of the 2,3- and 3,4-unsaturated derivatives from the same 3-O-sulfonyl derivative is discussed. PMID- 8407590 TI - Synthesis and biological activity of 3-(N-substituted pyridinium-4-thiomethyl)-7 alpha-formamido cephalosporins. AB - The synthesis and antibacterial activity of a series of 3-(1-substituted pyridinium-4-thiomethyl)-7 alpha-formamido cephalosporins is described. All the derivatives showed good potency and stability to bacterial beta-lactamases. The antibacterial efficacy seen with the N-alkyl pyridinium substituents was enhanced by the introduction of a catecholic side chain at C-7 and by preparation of N (substituted amino)pyridinium derivatives. PMID- 8407591 TI - Synthesis and biological properties of some 3-[(N-substituted-amino)pyridinium-4 thiomethyl]-7-[2-(2-amino-thiazol- 4-yl)-2-(Z)-(methoxyimino)acetamido]ceph-3-em 4-carboxylates. AB - The synthesis and antibacterial activity of a series of beta-lactamase stable, broad spectrum 7-[2-(2-amino-thiazol-4-yl)-2-(Z)-(methoxyimino)acetamido]-cephalo sporins, characterised by a C-3-[N-(substituted-amino)pyridinium-4-thiomethyl] group, is described. Gram-positive and Gram-negative bacteria including extended spectrum beta-lactamase-producing strains were most susceptible to the N-amino- and N-methylamino derivatives (3a) and (3b); with the exception of Pseudomonas aeruginosa, (3b) was more active in vitro and in vivo than cefpirome or ceftazidime. PMID- 8407592 TI - Anisomycin and new congeners active against human tumor cell lines. PMID- 8407593 TI - Inhibition of acyl-CoA: cholesterol acyltransferase by helminthosporol and its related compounds. PMID- 8407594 TI - Angelmicins, new inhibitors of oncogenic src signal transduction. PMID- 8407595 TI - Bequinostatins C and D, new inhibitors of glutathione S-transferase, produced by Streptomyces sp. MI384-DF12. PMID- 8407596 TI - UCA1064-B, a new antitumor antibiotic isolated from Wallemia sebi: production, isolation and structural determination. PMID- 8407597 TI - Cyclophostin, acetylcholinesterase inhibitor from Streptomyces lavendulae. PMID- 8407598 TI - Studies on cell growth stimulating substances of low molecular weight. Part 3. Resorcinin, a mammalian cell growth stimulating substance produced by Cytophaga johnsonae. PMID- 8407599 TI - Studies on cell growth stimulating substances of low molecular weight. Part 4. Misakimycin, a mammalian cell growth stimulating substance produced by Streptomyces misakiensis. PMID- 8407600 TI - A speculation about the parallel ear asymmetries and sex differences in hearing sensitivity and otoacoustic emissions. AB - Hearing sensitivity and the prevalence of spontaneous otoacoustic emissions show parallel ear and sex differences in humans. Here it is proposed that these lateral asymmetries and sex differences may all result from differences in the 'strength' of the efferent inhibition delivered to individual cochleas. Specifically, it is proposed that the amount of efferent inhibition is relatively less in right ears and in females than in left ears and males. While it is unclear why or how ear asymmetries or sex differences would develop in the efferent system, by assuming that they do, one can begin to explain a number of basic facts of hearing. It is even possible that the lateral asymmetry in the efferent system may be related to the well-known cortical asymmetries that are believed to underlie speech perception, speech production, and other human abilities. PMID- 8407601 TI - Effects of K(+)-channel blockers on cochlear potentials in the guinea pig. AB - The effects of different K+ channel blockers, 4-aminopyridine (4-AP), tetraethylammonium (TEA) and quinine, on the various cochlear potentials were observed by the means of perilymph infusion. Each of the three blockers depressed the compound action potential. However, they exerted quite different effects on other cochlear potentials, especially comparing 4-AP, a fast K(+)-channel blocker, with two other blockers. 4-AP induced a significant increase in the magnitude of summating potential, while TEA and quinine decreased it; 4-AP showed no effect on the general endocochlear potential (G-EP, the EP value recorded directly from the scala media, SM) and the negative EP component (N-EP), while TEA and Quinine increased G-EP and decreased the absolute value of N-EP. They also exerted different effects on the EP changes induced by exposure to intense noise. The results indicate the different roles of different K(+)-channels in the generation of cochlear potentials. The relationship of the two components of EP (positive and negative) and the G-EP was discussed. PMID- 8407602 TI - A model for the mechanics of the stereociliar bundle on acousticolateral hair cells. AB - The stereociliar bundle on acousticolateral hair cells was modelled as a series of stiff rods (stereocilia), and springs (stereociliary links and rootlets). Predictions were made for the coupling of stimulus-induced deflections between the stereocilia on the hair bundle, and for the stretches of the different classes of link. Comparison of the results with the measured mechanical properties of hair bundles suggests that in the bullfrog sacculus the stiffness of a side link and a tip link are related to the rootlet's contribution to the stiffness of a stereocilium to deflection in approximately the ratio > or = 400:100:1. The results show that stretch of the tip links is closely related to the deflection of the hair bundle over a wide range of model parameters, while the stretch of the side links is more variable, and in some types of bundle the mean stretch of the side links may be zero or negative. The results are in accordance with the view that the tip links are in an appropriate position to detect the deflections, while the main role for the side links may be to couple the deflections between the stereocilia. The mechanical consequences of bundles of different configurations, as seen in different hair cell types, are investigated. PMID- 8407603 TI - Immunohistochemical localization of gamma-aminobutyric acid- and aspartate containing neurons in the guinea pig superior olivary complex. AB - The immunohistochemical localization of gamma-aminobutyric acid (GABA)- and aspartate-containing neurons was demonstrated in the guinea pig superior olivary complex, using purified antisera to GABA and aspartate, respectively. Medium sized oval GABA-containing neurons were found in the lateral superior olive, and bipolar medium-sized GABA-containing neurons were observed in the dorsal hilus of the lateral superior olive. Medium-sized to large round GABA-containing neurons were observed in the ventral nucleus of the trapezoid body. GABA-containing terminals were found throughout the superior olivary complex with the highest density in the ventral nucleus of the trapezoid body. Aspartate-like immunoreactivity was observed in medium-sized round or oval neurons in the lateral superior olive, small fusiform neurons in the ventral nucleus of the trapezoid body, fusiform medium-sized neurons in the medial superior olive and oval medium-sized neurons in the superior paraolivary nucleus and round medium sized neurons in the medial nucleus of the trapezoid body. Double staining method demonstrated that aspartate-containing neurons in the medial nucleus of the trapezoid body were surrounded by GABA-containing terminals. The present results suggested the possible origin of GABAergic and aspartatergic olivocochlear bundles. PMID- 8407604 TI - Immunohistochemical localization of S-100 protein in the saccule of the rainbow trout (Salmo gairdnerii R.). AB - The distribution of S-100-like immunoreactivity in the trout saccule (a presumed organ of hearing in fish) has been determined by means of immunohistochemistry. Within the sensory epithelium of the saccular macula, hair cells and myelinated saccular nerve fibers were found to be immunoreactive. Hair-cell immunoreactivity was relatively uniform throughout the macula except at the extreme periphery (rostral, caudal, ventral and dorsal), where staining was either decreased or absent. The immunoreactivity associated with myelinated nerve fibers was greatest at the peripheral edges of the nerve processes, a position corresponding to the location of Schwann cells. However, the nerve processes themselves (within and subjacent to the sensory epithelium), as well as cell bodies within the saccular nerve, were also immunoreactive. Thus, the immunoreactivity of the saccular nerve observed above the basal lamina can be attributed to the saccular nerve processes as well as to nerve-associated Schwann cells. Overall, the immunoreactivity displayed by hair cells was less intense than that associated with myelinated saccular nerve, as evidenced by a disappearance of signal in hair cells first, upon serial dilution of antibody. No S-100-like immunoreactivity was observed in supporting cells within the sensory epithelium or in epithelial cells in non sensory regions. A concentration of S-100-like immunoreactivity in hair cells and saccular nerve is suggestive of the presence of S-100 calcium-binding protein mediated activities in these cell types. PMID- 8407605 TI - Response of neurons in the lateral superior olive and medial nucleus of the trapezoid body to repetitive stimulation: intracellular and extracellular recordings from mouse brain slice. AB - The responses of neurons in the lateral superior olive (LSO) and medial nucleus of the trapezoid body (MNTB) to repeated electrical stimulation of the trapezoid body were investigated in a brain slice preparation of the mouse superior olivary complex. Brain slices, 400-500 microns thick, were cut in the frontal plane and were maintained for physiological recording in a bath of warm, oxygenated saline. Both intracellular and extracellular recordings were made with glass micropipettes filled with 4 M potassium acetate. Bipolar stimulating electrodes were placed on the trapezoid body ipsilateral and contralateral to the superior olive. Current levels were set so that an ipsilateral pulse elicited a single action potential in LSO and a contralateral pulse elicited a single action potential in MNTB. Trapezoid body fibers were then stimulated with trains consisting of 10 current pulses. Repeated stimulation at low rates resulted in a number of spikes equal to the number of current pulses. Pulse rate was then increased to determine the limits of response to repeated stimulation. In the MNTB, neurons were capable of following very high rates of stimulation without reduction in response probability. On the average, MNTB neurons responded with a probability of 0.9 at 667 Hz. In contrast, most LSO neurons were incapable of following high rates of stimulation. LSO neurons responded with a probability of 0.9 at 125 Hz. For some LSO neurons, application of strychnine (0.5 microM) to the bath increased the probability of firing at high rates. MNTB neurons were unaffected by strychnine. PMID- 8407606 TI - Excitation effects on LSO unit sustained responses: point process characterization. AB - LSO units recover from a spike discharge in a characteristic way, modeled by an intrinsic recovery function that is stimulus invariant up to a scaling factor and a shifting constant. Data analysis shows that the effect of increasing excitatory stimulus level can be described by amplifying the intrinsic recovery function and by shifting it toward shorter intervals. The shifting process secondarily interacts with the absolute deadtime to produce the response characteristics of the three LSO unit types. Decreased excitation is clearly distinguished from inhibition, which affects the scaling, but not the time origin, of the recovery. We conclude that both excitatory and inhibitory stimulus levels are encoded in the timing of LSO unit discharges. PMID- 8407607 TI - Neuronal degeneration of primary cochlear and vestibular innervations after local injection of sisomicin in the guinea pig. AB - This paper reports on a dynamic study of the morphological changes within the cochlear and vestibular ganglia of the guinea pig after local application of Sisomicin in the inner ear. The treatment leads to a rapid, complete and irreversible destruction of the sensory cells in the cochlear and vestibular neuroepithelia. A progressive degeneration of the type I and type II afferent neurons, presenting a decreasing gradient from the base towards the apex of the cochlea, is rapidly observed and becomes almost complete as early as 15 days after the peripheral injury. Five months after the treatment the spiral ganglion cells have almost completely disappeared. At this time the vestibular ganglion cell density appears normal but the neurons exhibit important signs of alteration. Such damage to the cochlear and vestibular afferent neurons may result from either retrograde neuronal degeneration and/or direct neurotoxic effect of the drug. Thus the combination of the two mechanisms could lead to neuronal losses in spiral and Scarpa's ganglia after the local aminoglycoside intoxication of the inner ear. The difference in the time course of degeneration for these two afferent ganglia could be due to their specific susceptibilities or to their different anatomical locations. PMID- 8407608 TI - Spontaneous otoacoustic emissions: measurement and data. AB - Sounds from the ear canal were measured and then analyzed off-line. A peak picking algorithm located spectral maxima which might be designated as spontaneous otoacoustic emissions (SOAEs). The output from a 0.5-cc syringe, used to stimulate the volume of the ear canal, was also measured, analyzed and used to approximate the false-alarm rate of the measurement system. SOAE prevalence estimates depended on the false-alarm rate, just as the hit rate in a yes-no task does. With a false-alarm rate of zero, trends that have been found to be significant in the pooled results of other surveys were replicated: more SOAEs in the right ear, more SOAEs in females, and increased probability that the contralateral ear has an SOAE if the ipsilateral ear has an SOAE. In addition, many SOAEs failed to be detected in consecutive spectral analyses because they fluctuated in level. PMID- 8407609 TI - Sensory adaptation in frog vestibular organs. AB - Adaptation, i.e., the decrease with time in sensory units' afferent discharge to a constant stimulus, appears to be a common feature of the receptors belonging to acoustico-lateralis system: However, the mechanisms underlying this process are still a matter of debate. The present experiments demonstrate that sensory adaptation to both mechanical and electrical stimuli can be nearly suppressed after perilymphatic ouabain administration. This clearly indicates that the K+ homeostatic mechanisms [Valli et al., (1990) J. Physiol. (London) 430, 585-594] which control the K+ concentration gradient at both ends of vestibular hair cells play a predominant role in this process. The possible importance of different K+ dependent mechanisms in hair cell adaptation is discussed. PMID- 8407610 TI - Hair-bundle stiffness dominates the elastic reactance to otolithic-membrane shear. AB - Efficient transduction by acousticolateralis organs requires that a stimulus force principally deflect hair bundles, rather than flex other structural elements. Hair bundles might therefore be expected to provide a large fraction of the impedence to shear motions of otolithic membranes and other accessory structures. We measured the stiffness for shear motions of the bullfrog's saccular otolithic membrane, and determined the stiffness due to a single hair bundle and its associated extracellular filaments; this component is termed the elemental stiffness. Stiffness measurements were made by displacing the base of a flexible probe whose tip was coupled to the otolithic membrane, and simultaneously measuring the flexion of the probe and the displacement of the membrane. The average elemental stiffness, about 1350 microN.m-1, only modestly exceeded the stiffness of individual hair bundles. The hair bundles therefore provide the dominant component of stiffness in the bullfrog's sacculus, and thus account for a significant component of impedance to otolithic-membrane shear. As a corollary, stiffness changes or active movements in hair bundles should influence the mechanical responses of this and other receptor organs. PMID- 8407612 TI - Technology 1, scholarship 0. PMID- 8407611 TI - A realizable cochlear model using feedback from motile outer hair cells. AB - A physically realizable form of a recent cochlear model using feedback forces from motile outer hair cells [Geisler (1991) Hear. Res. 54, 105-117] has been developed. The model was computer-simulated in the frequency domain (necessarily linear). Its responses to pure tones are very realistic in terms of sharpness (Q10s of 3-5) and in terms of tip-to-tail ratios (50-60 dB). These large tips are due to the feedback forces, which act as negative resistances (energy-supplying elements) over restricted spatial ranges. Nyquist-criterion analysis indicates that the model is stable. The spatial patterns of the model's output also bear qualitative resemblances to several other phenomena observed in cochleas, both living and excised. PMID- 8407613 TI - Circumcision and health among rural women of southern Somalia as part of a family life survey. AB - A study of 859 rural women in 16 semipastoralist and semiagricultural villages in Southern Somalia reaffirmed the special significance of female circumcision as a source of full womanhood and an instrument for the control of female sexuality in Somalia. Although condemned in the West, this strongly embedded tradition enjoys unrestrained privilege in one of Africa's predominantly Islamic pastoral societies. The author reviews and analyzes the age-old tradition of circumcision, paying attention to the accompanying health problems as part of the Family Life Center's program. The Family Life program aims at improving the health conditions of rural women in Somalia and increasing their access to educational and economic opportunities. PMID- 8407614 TI - Female circumcision/genital mutilation: culturally sensitive care. AB - The ancient ritual of female circumcision/genital mutilation is still a prevalent custom in some African countries. Because of increasing immigration of African families to Western nations, the practice has become an issue for health care providers, who may not be aware of the practice and its sequelae. Using a questionnaire administered by our Somalian research associate, we surveyed women in schools of nursing and education in Somalia, identifying the complications of circumcision, acceptable interventions for these complications, and caregivers considered appropriate by the respondents. By becoming aware of the practice and of which interventions are acceptable and unacceptable, health care providers can offer more comprehensive and culturally sensitive care. PMID- 8407615 TI - Nigerian urban married women's perceptions of exposure to secondary tobacco smoke. AB - We investigated urban, married Nigerian women's knowledge of the health risks associated with secondary tobacco smoke, their attitudes toward this problem, and the preventive efforts they took when they or their children were exposed involuntarily to secondary tobacco smoke. Two hundred forty-nine women were surveyed with a structured, written questionnaire. The results indicated that these predominantly well-educated and professionally employed Nigerian women had only a fair knowledge of the adverse health effects of exposure to secondary tobacco smoke. Although their attitudes toward exposure to secondary tobacco smoke were generally quite good, they were reluctant to take preventive measures in public. The most prominent factors relating to knowledge and attitudes were age, educational level, and smoking status. Attitudes proved to be a stronger predictor of preventive efforts than knowledge. PMID- 8407616 TI - Sex differences in preferences for ideal female body shape. AB - The investigator aimed to replicate on an African sample Fallon and Rozin's (1985); Huon, Morris, and Brown's (1990); and Tiggemann and Pennington's (1990) findings of sex differences in female body preferences. Males and females were asked to identify their ideal female body shape and the one they believed men and women in general would prefer. A simple analysis of variance and t test demonstrated a discrepancy between women's actual and preferred body sizes, while the males knew the females' preferred size. These findings support the previous studies. The female undergraduates' and adult women's consistently negative evaluations of their own body compared with their ideal or preferred body are explored and found to result from being subjected to implicit pressures toward thinness not suffered by their male counterparts. Recommendations for future research are made. PMID- 8407617 TI - The ethnographic context of illness among single-women-headed households in rural Peru. AB - Peru is undergoing many deleterious economic and social changes, and the health consequences for families headed by single women is of special concern. However, not all single mothers' families may be at similar risk of morbidity. My purpose in the present study was to determine whether variation in illness prevalence was associated with ostensibly small differences in socioeconomic resources and physical living conditions in areas like Peru. I used both qualitative and quantitative methods in this study. The sample included single-women-headed households from the rural Nunoa District, located in the southern Peruvian Andes (N = 22 families with 90 individuals). Quantitative cross-sectional survey results suggested that the mother's formal education, the availability of a latrine, drinking water contamination, the gender ratio of the household, and the quality of the social support network were key risk factors. They statistically predicted 35-91% of the morbidity variance between families (p < or = .03-.0001). I illustrate these findings in three ethnographic case studies. The case studies show how the degree of illness among single-women-headed families was affected by small differences in their social and physical living conditions. PMID- 8407618 TI - Birth weight and smoking practices during pregnancy among Mexican-American women. AB - The smoking practices of a national sample of Mexican-American mothers and the resulting effects of those practices on birth weight were examined. Data were from the Hispanic Health and Nutrition Examination Survey conducted by the U.S. National Center for Health Statistics during 1983-1984. We found that 24% of the mothers had smoked during their pregnancy, with a mean of 11 cigarettes per day. Infants of these women weighed 101 g less at birth than did infants of nonsmoking mothers and had a low birth weight rate of 8.0% compared with the 5.1% low birth weight rate for the sample as a whole. Multiple regression results indicate a 7.4 g decrease in birth weight for each cigarette smoked per day during pregnancy. Cultural factors that promote a low birth weight rate for Mexican-Americans that is comparable to that of non-Hispanic whites despite increased rates of poverty and inadequate health care do not protect against the insidious effects of cigarette smoking. PMID- 8407619 TI - Swedish students' attitudes toward abortion. AB - The Swedish abortion legislation of 1975 gave women the right to make a decision about abortion before the end of the 18th week of pregnancy. The number of abortions is rising in Sweden as a chosen method of birth control. The attitudes of students toward abortion were studied in 1986-1987. A questionnaire containing items on how sex education is taught, the anatomy and physiology of reproduction, contraceptives, sexually transmitted diseases, and legal abortion was answered by 421 high school students. Results pertaining to the students' attitudes toward abortion are reported. Two thirds of the students believed that the decision about an abortion should be made by the man and woman together. Nearly all respondents believed that abortion should not be considered a method of birth control. These results may be considered a guide for interventions to prevent the need for abortion. One fourth of all pregnancies in Sweden terminate in abortion. The students in the present study thought of abortion as a solution. Authors studying samples with different cultural backgrounds have reported similar attitudes. PMID- 8407620 TI - Incidence of sexually transmitted diseases and Pap smear results in female homeless clients from the Chicago Health Outreach Project. AB - Homeless persons have difficulty gaining access to health care. In 1985 the Chicago Health Outreach Project was created to improve their access to health care. Staff and client reviews indicated that female homeless clients required increased outreach efforts. Consequently, a mobile women's health unit was developed in 1990. Review of 128 records of 104 female homeless clients indicated that 30% of Pap smears done were abnormal, with atypia (14%) and inflammation (10%) the most common findings. The incidence of chlamydia, gonorrhea, and trichomoniasis was 3%, 6%, and 26%, respectively. There was a significant association between the presence of trichomoniasis and an abnormal Pap smear. These results emphasize the importance of providing regular gynecological care to homeless women. Research is needed on the implications of abnormal Pap smears and sexually transmitted diseases for this population of female homeless clients. PMID- 8407621 TI - Rural women's knowledge of and attitudes toward acquired immune deficiency syndrome. AB - We surveyed the knowledge of and attitudes about acquired immune deficiency syndrome (AIDS) of 236 women living in a rural community. The design of the study was descriptive, using the AIDS Knowledge and Belief Survey. There was a significant positive correlation between attitude scores and knowledge scores. Women who had read about AIDS had significantly higher knowledge scores than women who had not read about AIDS, but there were no significant differences between the attitude scores of women who had and had not read about AIDS. PMID- 8407622 TI - The ethics of politically correct behavior. PMID- 8407623 TI - From gynecology to GYN-ecology: developing a practice-research agenda for women's health. AB - Women's health as an area of specialization has moved away from a focus on gynecology to a focus on GYN-ecology, meaning a concern for the fit between the woman and her environment. To be concerned about the fit between person and environment demands an understanding of the extent to which individual behavior is constrained or facilitated by contextual factors: Do larger sociopolitical conditions exist in which a woman can perform the activities of daily living in a way that maximizes her sense of well-being? Seven factors are explored to illustrate the many ways in which women's health has been traditionally constrained by contextual factors. Suggestions for a comprehensive practice research agenda for women's health are then presented. PMID- 8407624 TI - Women, health, and knowledge: travels through and beyond foreign parts. AB - The meanings of health for women as a social group are discussed. It is argued that women's perspectives on health give central importance to the concepts of self-determination, coping, power, and control. Health for women is bound up with their experiences in everyday life. Women are also primary health care providers. Medicine, however, dominates women's bodies in ways that conflict with women's own perspectives on their health: In "developed" societies, medical intervention now poses a major threat to women's health. By concentrating on quantitative methodology, most researchers studying women's health have ignored these important meanings of health to women. PMID- 8407625 TI - Care--beyond virtue and command. AB - The academic turn in the field of health care has placed the study of health within the confines of a logocentric or scientistic framework. This development has led to the devaluation of many issues important to professional practice. One of these is the issue of care. In this article, different ways to enhance caring qualities are discussed. It is shown that the key to care lies not within abstract theories and principles, but rather in reflecting over our own life world experience. PMID- 8407626 TI - Women, health, and technology. AB - Some factors that determine the development and diffusion of medical technologies are discussed. The use of technology in pregnancy and labor cannot be seen as a response to user needs or the result of societal planning alone. Often technologies are developed and diffused without any evaluation of their risks and benefits. The relations between medical research and industrial interests are focused on to explain the development, and against this background user interests and women's strategies as users and providers of health care are discussed. PMID- 8407627 TI - Strategies for empowering women's voices in the medical culture. AB - Power and knowledge are closely connected, and this is no less true for the medical profession than it is for any other sphere of life. Knowledge is constructed by voice. Unfortunately, women's voices are often silent in the factory where medical knowledge is produced. Medicalization and ignoring are symptoms of the medical oppression of women's voices. Empowerment of women's voices at various levels within the medical culture is essential for influence and social reconstruction. In this regard, a research project on alternative approaches to physician-patient communication as well as research methods is presented. Knowledge constructed from women's voices will be neglected by medical power unless it can hold up to systematic investigative procedures. Sensitive and sensible research from women's voices requires methodology that preserves women's messages and transforms them into medical knowledge. Such efforts may ultimately lead toward construction of a feminine medical epistemology--a medical knowledge that reflects women's reality. PMID- 8407628 TI - Women's health status in Africa--environmental perspectives from rural communities. AB - Health research used to be the exclusive domain of clinicians and medical specialists, who focused attention on the biomedical causes of disease. Socioeconomic and environmental considerations that have important bearing on the ill health of rural African women were rarely integrated with the methodology constructed to investigate disease patterns. However, it is becoming increasingly clear that physical environmental factors and malnutrition have important effects on women's health in rural Africa. I validated this assumption in an empirical study of 441 people (n = 294 women) in 15 different rural localities in Ghana. Apart from women-specific problems relating to biological health needs during pregnancy, childbirth, and lactation, sexually transmitted diseases, abortion, and mental health, environmental factors had a great impact on women's well-being in the study area. Sixty-two percent of the women reported that the endemic disease malaria is the most prevalent disease as far as they were concerned. Other community and household health hazards were found; for example, cooking over an open fuel wood stove resulted in an almost 50% greater chance of stillbirth among pregnant women. PMID- 8407629 TI - Feminist critique. PMID- 8407630 TI - The new woman: health-promoting and health-damaging behaviors. AB - Compared with their mothers, contemporary women are better educated, live alone more of their adult lives, and participate in the labor force more consistently throughout their lives. Most studies of the New Woman have focused on the effects of the configuration of her roles on her health and health-related behavior. Few have considered both the influence of women's roles and the broader context of their lives on their health. Moreover, investigators have devoted little attention to the health behavior patterns of the New Woman, despite the demonstrable impact of health-related behaviors on mortality and morbidity. The present study was designed to test models relating women's roles, gender role norms, social demands and resources, and health-promoting and -damaging behaviors. Women who resided in middle-income and racially mixed neighborhoods (N = 659) were interviewed in their homes. The women were a mean age of 32.7 years and had an educational level of 14.2 years. Approximately 57% were married or partnered, and 76% reported some level of employment outside the home. Women who had fewer life stressors and were better educated performed more health-promoting behaviors. Women who experienced more stressors, were less well educated, had more contemporary attitudes toward women, were not partnered, or experienced depressed mood engaged in more health-damaging behaviors. These results underscore the importance of considering women's social context in health promotion efforts and as a deterrent to health-damaging behaviors. PMID- 8407631 TI - Institutionalizing women's oppression: the inherent risk in health policy that fosters community participation. AB - Current Canadian health policy is based on the implicit assumption that women are available to provide care in the home to the dependent, the ill, the elderly, and the physically and mentally disabled. Women are socialized from birth to accept caring roles within a traditional family structure, and current societal expectations and social policy reinforce this value system. Women's health can only be understood within the context of their lived experience of social inequity, medicalization, and family caregiving. Health care professionals are complicit in sustaining women's oppression by reinforcing these institutions of social control. For health policy to be responsive to women's needs, it must be based on research that considers the social complexity of ordinary women's lives. PMID- 8407632 TI - The experiences of midlife daughters who are caregivers for their mothers. AB - A study was conducted of the experiences of midlife daughters who are caregivers for their aging mothers. I recruited participants through a daughters-of-aging parents program at the Women's Resource Centre in Vancouver, British Columbia. The phenomenological research method was used to elicit midlife daughters' descriptions of their caregiving experiences and the emotions generated by these experiences. The findings elucidated a development framework--the Continuum of Care--along with daughters vacillated during their caregiving experiences. Health care practitioners can use the Continuum of Care framework to identify triggers to assist daughters to develop healthy caregiving relationships with their mothers. PMID- 8407633 TI - Midlife Baby Boom women compared with their older counterparts in midlife. AB - A stratified random sample of midlife Baby Boom women, ages 35-42, was compared with a group of older midlife women, ages 43-55. The sample consisted of 992 women who had graduated between the years of 1955 and 1975 from a large university in southern California. Measures of anxiety and life satisfaction were administered to each year group. This served to control for the effect of age and was helpful in explaining whether the differences between the groups were age related or the result of socialization. The Baby Boom cohort of women, born after January 1, 1946, have only recently entered the ranks of midlife. They have been recognized as unique and less traditional than their older counterparts with regard to sexual freedom, career choices, and educational opportunities. Similarities and differences were found between the two groups of women. However, the results clearly revealed that the effects of socialization, rather than age, yielded the major significant variables influencing a feeling of well-being in both cohorts of midlife women. PMID- 8407634 TI - Interrupted work histories: retired women telling their stories. AB - The issue of women and work is a contemporary one which affects the private as well as the public sphere of women's lives. The social conditions which affect the present cohort of retired older women, have their roots in their work lives. In this article, we analyzed the oral histories of 7 retired women using a modified grounded theory approach to explore the long-term effects of having interrupted their work histories in order to care for family members. The core variable that emerged was the "valuing process" that occurred when these older women relieved their past experiences. We defined this as the relative merit that each participant attributed to her lifetime work achievements. PMID- 8407635 TI - An ecological model of group well-being: implications for health promotion with older women. AB - One promising context for health promotion with older individuals is the continuing small group, meeting on a regular basis for participation in health and wellness activities. As the facilitator of such a group, I have been conducting research using ethnographic and constant comparative methods on the dimensions of group well-being. Three core themes are described: (a) group as ritual, (b) group as celebration, and (c) group as community. Findings extend my previous report (Ruffing-Rahal, 1989) of constructs of well-being among older persons by delineating the small-group properties of well-being. PMID- 8407636 TI - Factors related to older women's recovery from hip fractures: physical ability, locus of control, and social support. AB - Our purpose in conducting this study was to identify the nature of care provided by the informal support network available to older women with hip fractures and to examine the relationships among prior functional ability, locus of control, and reliance on social supports in predicting recovery from hip fractures. One hundred one women who had recently sustained a hip fracture participated in a structured interview process. Overall, the women received more frequent help from their spouse, children, and friends at the time of the interview than during the month before their hip fractures. Women who had higher activities of daily living capabilities prior to the fracture, had a greater internal orientation, and used fewer formal services reported higher levels of current physical functioning. PMID- 8407637 TI - Perinatal loss: a framework for practice. AB - Perinatal death is a crisis for women and their families. By using a conceptual framework to guide care, caregivers can facilitate grief and help the healing process. The framework described in this article is centered on the key concept of meaning or significance of the loss to the woman, her partner, family, friends, community, and society. The loss may encompass many meanings, and the meanings may differ from person to person. Sociocultural, spiritual, psychological, developmental, and physiological variables are presented. Goals of intervention include promotion of personal choice; support for decisions made; and promotion of dignity and respect for the woman, baby, and family. Intervention modes are supportive, informational, and facilitative. PMID- 8407638 TI - Edwin Bret Hart, 1874-1952: a brief biography. PMID- 8407639 TI - Effects of supplementing growing steers with high levels of partially hydrogenated tallow on feed intake, digestibility, live weight gain, and carcass characteristics. AB - Effects of supplementing growing steers consuming forage-based diets with high levels of partially hydrogenated tallow on feed intake, digestibility, live weight gain, and carcass characteristics were determined in this study. In Exp. 1, Holstein steer calves were fed bermudagrass hay alone (Control) and with a ground corn-based concentrate at .95% BW (DM basis) plus partially hydrogenated tallow at 0 (Basal), .33 (low fat; LF), or .67% BW (high fat; HF) in a Latin square. Total DMI was increased by concentrate supplementation and was lower for HF than for LF (P < .05; 3.18, 3.83, 4.25, and 3.17 kg/d for Control, Basal, LF, and HF, respectively). In Exp. 2, grazing Angus x Hereford steers (270 kg +/- .4 initial shrunk BW) were fed the same supplements as in Exp. 1 for 84 or 98 d and slaughtered. Live weight gain was increased (P < .05) by concentrate supplementation (1.01, 1.34, 1.41, and 1.30 kg/d for Control, Basal, Low, and High, respectively). The concentration of total lipids in longissimus muscle was 2.51, 2.53, 3.05, and 3.03% of wet tissue for Control, Basal, LF, and HF, respectively (Basal > the mean of LF and HF; P < .07); the proportion of palmitic acid in total fatty acids was similar among treatments. Fat supplementation did not markedly affect sensory, taste, or tenderness characteristics of longissimus muscle. In conclusion, supplementing grazing beef steers with high levels of partially hydrogenated tallow, with slaughter at approximately 400-kg shrunk BW, tended to increase fat in longissimus muscle without altering the palmitic acid level in fatty acids, although sensory, taste, and tenderness characteristics were not modified. PMID- 8407640 TI - Effects of ivermectin on reproductive functions in ewes. AB - In response to producers' concerns about possible detrimental effects of ivermectin on ewes during the breeding season, an evaluation of its effects on endocrinological, physiological, and behavioral measures of reproductive performance was made. Twenty cycling ewes were randomly assigned on d 10 (d 0 = estrus) to receive a single recommended oral dose of 200 micrograms/kg of BW of ivermectin or a control volume of water. Twelve hours after treatment, ewes received a luteolytic injection of 5 mg of PGF2 alpha and were introduced to fertile rams. On d 10 after mating, laparoscopies were performed to assess ovulation rate, and on d 18 conceptuses were surgically recovered. Blood samples were collected during the 92-h interval beginning immediately before ivermectin and control treatments and were assayed for LH. It was determined that 1) interval from PGF2 alpha-induced luteal regression to a) onset of estrus (36.1 +/ 1.4 vs 36.3 +/- 1.4 h) and b) onset of the preovulatory surge of LH (38.2 +/- 2.6 vs 44.2 +/- 2.6 h), 2) magnitude of the surge of LH (275.6 +/- 38.9 vs 199.8 +/- 38.9 ng/mL), 3) duration of the surge of LH (9.4 +/- .4 vs 9.0 +/- .4 h), 4) area under the curve of the surge of LH (54,321 +/- 7,419 vs 38,138 +/- 7,419 arbitrary units), 5) ovulation rate (2.1 +/- .4 vs 2.0 +/- .3 ovulations/ewe), 6) pregnancy rate (8/10 vs 5/10 ewes pregnant), and 7) conceptuses per ewe (1.75 +/- .37 vs 1.60 +/- .33) did not differ (all P > .1) between ivermectin- and control treated ewes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407641 TI - Comparative feeding value of wood sugar concentrate and cane molasses for feedlot cattle. AB - Two feedlot cattle growth performance trials and a metabolism trial were conducted to evaluate the feeding value of cane molasses (CM) compared with wood sugar concentrate (WSC). The CM contained (as-fed basis) 73% DM, 8.6% ash, 48% total sugars, and 79.5 degrees Brix. The WSC contained (as-fed basis) 61% DM, 12.2% ash, 38.9% total sugars, and 29% lignosulfonate. When supplemented at 4% of diet DM, treatment effects on ruminal and total tract OM digestion and feedlot performance were similar. When supplemented at 8% of diet DM, ADG was similar across treatments, whereas feed efficiency tended (P < .10) to favor the CM diet. The NEm value of the diet was 2% higher (P < .01) with CM supplementation. It is concluded that WSC may be a useful alternative to molasses in diets for cattle. The feeding value of WSC seems to exceed that based on its hydrolyzable sugars content. Given that the NEm and NEg values (DM basis) for CM are 1.70 and 1.08 Mcal/kg, respectively, the corresponding values for WSC are 1.25 and .68 Mcal/kg, respectively. PMID- 8407642 TI - Influence of processing on the feeding value of oats for feedlot cattle. AB - Ninety-six crossbred steers (312 kg) were used in a 118-d feedlot growth performance trial to determine the relative effects of degree of steam processing on the feeding value of oats in a 90% concentrate finishing diet. Treatments were 1) steam-flaked corn (SFC, density = .28 kg/L); 2) dry-rolled oats (DRO, density = .36 kg/L); 3) steam-rolled oats, coarse flake (SRO-C, density = .33 kg/L); and 4) steam-rolled oats, thin flake (SRO-T density = .17 kg/L). Feed intake was similar (P > .10) across oats treatments. However, ADG was lower (13.2%, P < .01) and feed/gain was higher (11.1%, P < .05) for SRO-T than for SRO-C diets. Diet NEm was 7.2% lower (P < .10) for SRO-T than for SRO-C diets. Daily weight gain was greater (9.3%, P < .01) and feed intake and feed/gain were lower (13.1 and 20.8%, respectively; P < .01) for SFC than for oats treatments. Diet NEm and NEg were higher (20.0 and 26.5%, respectively; P < .01) for SFC than for oats treatments. Dressing percentage was 2.5% lower (P < .01) in steers fed the oats diets than in those fed the SFC diet. Four Holstein steers (232 kg) with cannulas in the rumen and proximal duodenum were used in a 4 x 4 Latin square experiment to evaluate treatment effects on characteristics of digestion. Ruminal digestibility of OM, starch, and ADF were similar (P > .10) for oats treatments, averaging 58.2, 93.5, and 9.6%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407643 TI - Effect of source of supplemental nitrogen on the utilization of citrus pulp-based diets by sheep. AB - Two trials were conducted to examine the influence of N enrichment of dried citrus pulp (DCP) through ammoniation vs urea supplementation on digestion and feeding value of a DCP-based diet for sheep. Treatments were 1) ammoniation of DCP with urea (CP-U), 2) ammoniation with ammonium hydroxide (CP-A), 3) supplementation with urea, and 4) supplementation with horse bean (HB). Diets were isonitrogenous (1.8% N). In Trial 1, treatment effects on characteristics of digestion were evaluated in a 4 x 4 Latin square experiment using four rams. Source of supplemental nonprotein N (NPN) did not influence (P > .10) ruminal OM digestion. Net microbial N synthesis, N absorption, N retention, and microbial efficiency were greater (P < .05) for the CP-U than for the CP-A or urea diets. Apparent N digestibility of CP-A was 14 to 25% lower (P < .05) than that of the other diets supplemented with NPN. In Trial 2, treatment effects on lamb growth performance were evaluated in a 105-d growing-finishing trial involving 32 Sardi lambs (19 kg). Intake was similar (P > .10) across treatments (1.13 kg of DM/d). Daily gain of the urea-fed lambs (182 g) was higher (P < .05) than that of CP-U- or CP-A-fed lambs (138 g). Feed efficiency and dietary NE values were similar (P > .10) among diets containing NPN. Growth performance seemed to be more directly affected by intake, rather than N economy. Enrichment of DCP through ammoniation with urea was a superior method of NPN supplementation with respect to efficiency of N utilization. However, the ultimate response to the potential benefits of such treatment may be seen only if N is truly limiting performance. PMID- 8407644 TI - Reproductive and maternal performance of rotational three-breed, and inter se crossbred cows in Florida. AB - Reproductive, calf growth, and cow weight data were collected during a 16-yr period in southern Florida. Data included 1,767 calves at weaning from purebred, F1, backcross, F2, and three-breed crossbred cows of the Angus (A), Brahman (B), and Charolais (C) breeds. The purebred cows primarily produced purebred calves, the F1 and F2 Cows produced inter se crossbred calves, and the backcross cows produced 3/8:5/8 calves. The 1/2 C:1/4 A:1/4 B three-breed crossbred cows were mated to A series, whereas three-breed crossbred dams with breed compositions of 1/2 B:1/4 A:1/4 C, and 1/2 A:1/4 B:1/4 C were mated to C and B sires, respectively. For analysis of additive breed and heterosis effects, pregnancy rate and cow weight were considered to be traits of the dam alone. The A additive breed effect increased (P < .05) pregnancy rate but reduced (P < .001) cow weight. The effects of AB, AC, and BC heterozygosity all increased pregnancy rate, but the advantage was greater for the crosses that involved B. Both AB and BC heterozygosity increased (P < .001) cow weight, whereas AC did not. The additive direct effect of B and C increased birth weight (BWT) and weaning weight (WWT). The additive maternal effect of B reduced BWT. The direct effect of AB heterozygosity increased BWT and WWT, and that of BC increased only WWT (P < .001). A comparable pattern was observed for maternal heterosis on weight traits. A much smaller effect of AC maternal heterosis on WWT was found. Pregnancy rates of F1, backcross, and three-breed dams were similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407645 TI - Correlations for genetic expression for growth of calves of Hereford and Angus dams using a multivariate animal model. AB - The purpose of this study was to estimate the correlation between the expression of genes from sires in purebred and crossbred progeny (rPC) and in Hereford and Angus F1 calves (rHA). Performance traits were weights at birth, 200 d, and 365 d. Progeny from Hereford, Polled Hereford, and Angus bulls mated to Hereford or Angus cows were used to estimate rPC. Progeny from Charolais, Shorthorn, Simmental, Limousin, Maine-Anjou, Chianina, Gelbvieh, Tarentaise, and Salers bulls mated to Hereford or Angus cows were used to estimate rHA. Performances in purebreds (P) and crosses (C) or in Hereford (H) and Angus (A) F1 calves were treated as separate traits. A multivariate animal model with birth year-cow age sex subclasses, random correlated direct and maternal additive genetic effects, and maternal permanent environmental effects was used. Separate analyses were done by breed of sire. A derivative-free algorithm was used to obtain REML estimates of (co)variance components. Weighted averages across breeds of estimates of heritability for P, C, H, and A were, respectively, .61, .51, .47, and .40 for birth weight, .41, .46, .37, and .34 for weaning weight, and .50, .49, .42, and .46 for yearling weight. Estimates of rPC ranged from .88 to .97, .55 to .94, and .68 to .86 for weights at birth, 200 d, and 365 d, respectively. Estimates of rHA ranged from .43 to .99, .56 to .95, and .50 to .98 for weights at birth, 200 d, and 365 d, respectively. Weighted averages of estimates of rPC and rHA across sire breeds were, respectively, .93 and .85 for birth weight, .77 and .73 for weaning weight, and .76 and .86 for yearling weight. These results indicate that ranking of sires producing purebreds or crosses, or crossbred calves from different breeds of dams, is approximately the same for birth and yearling weights, but some reranking might occur for weaning weight. PMID- 8407646 TI - Considerations on genetic connectedness between management units under an animal model. AB - Connectedness among management units (e.g., herds or regions) is of concern in genetic evaluation. When genetic evaluation is under an animal model, connections occur through A, the numerator relationship matrix. It is argued that the most appropriate measure of connectedness is the average prediction error variance (PEV) of differences in EBV between animals in different management units. It is shown that PEV of differences is influenced by average genetic relationship between and within management units, which in turn affects the variances of estimates of differences between management unit effects. When PEV of differences cannot be computed, use of one of three alternative measures is proposed; the gene-flow method that measures the exchange of genes between management units, measurement of genetic drift variance based on average relationships between and within management units, and measurement of the variance of estimated differences between management units effects. These were correlated with PEV of differences in a test simulation. The gene-flow method, which is simplest to compute, had the lowest correlation (.671). The drift variance and variance of management unit effects methods were highly correlated with PEV of differences (.924 and .995, respectively). PMID- 8407647 TI - The influence of maternal effects on accuracy of evaluation of litter size in swine. AB - The influence of maternal genetic effects on response to selection was examined by stochastic simulation. The selection process of a closed herd of 120 sows, with 24 boars entering the breeding herd each year, was simulated over 10 yr. Effects of different magnitudes of maternal heritability (0, .025, .05), genetic correlations between maternal and direct effects (0, -.5, -.9) and evaluation models (with or without maternal effects, referred to as complete or incomplete model) on response to selection, accuracy of evaluation, prediction error variance (PEV), bias, and mean squared error (MSE) were analyzed for litter size with a direct heritability of .10. Directional selection of replacement animals was on EBV of direct effects for first-parity litter size under an animal model. Using a complete animal model with maternal effects, response in direct genetic effects increased with magnitude of maternal heritability (0 to .05) from 2.22 to 2.32 pigs after 10 yr, when there was no correlation between direct and maternal effects. Additionally, a positive maternal response was achieved (with maternal heritability > 0), although no selection was on maternal EBV. Reduction in direct response due to negative genetic correlations between direct and maternal effects was up to 18% after 10 yr of selection. More important was the negative maternal response, which was up to -1.27 pigs after 10 yr for a genetic correlation of .9. Consequently, the overall genetic merit (maternal plus direct) was reduced up to 77% compared with when maternal and direct effects were genetically independent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407648 TI - Genetic parameters for carcass traits estimated from Angus field records. AB - The American Angus Association has sponsored a carcass evaluation since 1974. The carcass data collected as a part of this program are used by the association to conduct a biannual sire evaluation for carcass merit. This paper presents age adjustment factors and genetic parameter estimates for carcass traits to be used in the Angus carcass genetic evaluation program. Because of the large range in slaughter ages, age classes were defined as all those animals slaughtered at an age of < or = 480 d and those with a slaughter age > 480 d. Linear and quadratic partial regressions on slaughter age for hot carcass weight (HCW), USDA marbling score (MS), 12th rib longissimus muscle area (LMA), and 12th-rib fat thickness (FT) were estimated within sex and age class. Quadratic age regressions were not significant, nor was the linear age regression coefficient for FT in steers in the > 480-d age class. Heritability estimates for age-constant HCW, MS, LMA, and FT were .31, .26, .32, and .26, respectively. The estimated genetic correlation (rg) between HCW and LMA was .47. The estimated rg between HCW and FT was .38 and between MS and FT was -.13. The linear genetic trends for CWT and LMA were significantly positive at .414 kg/yr and .075 cm2/yr, respectively. The genetic trends for FT and MS were very small but significantly negative at -.004 cm/yr and -.003 units/yr, respectively. PMID- 8407649 TI - Relationship of beef sire expected progeny difference to maternal performance of crossbred daughters. AB - Cumulative milk production of crossbred daughters of sires of several breeds was estimated using weigh-suckle-weigh procedures. Pooled-across-breed analyses were conducted to determine, in retrospect, relationships of sire EPD values for milk and total maternal value to daughter milk yield and daughter's offspring weaning weight. The pooled coefficient for regression of daughter 214-d milk yield on sire milk EPD was 13.4 kg/kg (residual correlation was .14). The overall mean estimated milk yield was 1,262 kg, suggesting that a difference in sire milk EPD of 1 kg corresponded to a difference of approximately 1% in cumulative daughter milk yield. The pooled coefficient for regression of daughter's offspring 214-d weight on sire total maternal EPD was 1.18 kg/kg (residual correlation was .17). All regression and correlation coefficient stated above were significantly greater than zero. Breeders who use sire milk and total maternal EPD values as selection tools should expect such selection to be effective, on average, but should also expect that a substantial proportion of individuals or small groups may not rank as predicted. PMID- 8407650 TI - Performance, carcass composition, and blood hormones and metabolites of finishing pigs treated with porcine somatotropin in hot and cold environments. AB - Two experiments were conducted to assess the ability for recombinant porcine somatotropin (rpST)-treated pigs to perform and cope with the demands of hot and cold environments. In the first experiment, finishing pigs were exposed to either a thermoneutral (TN; 18 to 21 degrees C) or a hot environment (H; 27 to 35 degrees C) for 35 d. In the second experiment, pigs were exposed to a TN or cold environment (C; 5 to 15 degrees C). The rpST delivered by a 6-wk prolonged release system had no effect on ADG, whereas both H and C reduced ADG by 29.4 and 11.8%, respectively. In the first experiment, rpST-treated pigs consumed 17.6% less feed than control pigs, whereas rpST-treated pigs in H consumed 24.4% less feed than rpST-treated pigs in TN. Overall feed/gain ratios through the first 4 wk of both studies were improved by 21.8 and 14%, respectively, by rpST (P < .05) and were 24.3% poorer in C (P < .05) than in H. The changes in blood concentrations of pST, IGF-I, and IGF-II associated with rpST were not influenced by the different environments. Total body composition of rpST-treated pigs had increased amounts of protein (P < .05) and decreased amounts of fat (P < .05); H further reduced fat (P < .05). The C resulted in reduced protein content (P < .05). No evidence of thermal imbalance due to rpST was found as assessed by rectal temperature, respiration rate, and heat production estimated by indirect calorimetry and chemical analysis. PMID- 8407652 TI - Body weight gain is correlated with serum cholesterol at 8 weeks of age in pigs selected for four generations for low or high serum cholesterol. AB - We determined the relationship of BW at birth, weaning (4 wk of age), and 8 wk of age to serum total cholesterol (C), high-density lipoprotein-cholesterol (HDL-C), and triglycerides (TG) at 8 wk of age in pigs, from the fourth generation that had been selected for low (10 litters, 75 pigs, LC) or high (10 litters, 63 pigs, HC) C at 8 wk of age. Mean C concentration at 8 wk of age was 81 +/- 30 mg/dL for LC groups and 136 +/- 19 mg/dL for HC groups. Serum C, HDL-C, and TG concentrations were not correlated with birth weight, suggesting that the physiological factors that may cause reduced weight gain in older animals are not operative in newborn pigs. All three constituents were correlated (P < .05) with BW at weaning and at 8 wk. However, only 4% of the variation in weight at weaning and 7% at 8 wk could be explained by a relationship with serum TG. There was a positive correlation between C and BW at 8 wk (r = .46, P < .05), which was apparent within the subgroups of LC and HC females and LC males (r = .46, .48, .68, respectively); the correlation was low (r = .26) in HC males. PMID- 8407651 TI - Effects of growth hormone-releasing factor and(or) thyrotropin-releasing hormone on growth, feed efficiency, carcass characteristics, and blood hormones and metabolites in beef heifers. AB - The objective of this study was to determine the effect of long-term administration of a growth hormone (GH)-releasing factor analog (GRFa) and(or) thyrotropin-releasing hormone (TRH) on growth, feed efficiency, carcass characteristics, and blood hormones and metabolites in beef heifers. Crossbred heifers (n = 48; 345.9 +/- 2.8 kg) were divided into four equal groups: control (vehicle), 1 microgram of GRFa (human GRF 1-29 analog).kg BW-1.d-1, 1 microgram of TRH.kg BW-1.d-1, or GRFa + TRH. Daily s.c. injections continued for 86 d. Blood samples were collected from half of the heifers after injection on d 1, 36, and 78. On d 89, all heifers were slaughtered. Treatments did not affect (P > .05) ADG but GRFa + TRH decreased (P < .05) ADFI relative to all other treatments. Feed conversion efficiency tended (P < .10) to be improved in the groups given GRFa alone or TRH alone. Treatment with GRFa and(or) TRH did not affect carcass weight, dressing percentage, conformation score, backfat thickness, or weights of liver, kidneys, pituitary, and ovaries. The GRFa + TRH treatment reduced (P < .05) fat score and increased (P < .05) longissimus muscle area relative to other treatments. The GRFa treatments reduced (P < .05) the weight and fat percentage of the mammary gland and increased (P < .05) heart weight. Treatment with TRH alone failed to stimulate GH on d 1, 36, and 78. Treatment with GRFa alone increased (P < .05) GH above controls on d 36, whereas GRFa + TRH increased (P < .05) GH on d 1, 36, and 78. Treatment with GRFa alone increased (P < .05) IGF-I only on d 1, whereas GRFa + TRH was without effect on all days. Across sampling days, treatments had little effect on blood concentrations of insulin, triiodothyronine, nonesterified fatty acids, urea nitrogen, and glucose. The GRFa alone and GRFa + TRH decreased (P < .05) and TRH alone increased (P < .05) thyroxine concentrations. In conclusion, with the dose and administration regimen used, GRFa and(or) TRH yielded small but positive improvements in animal performance. PMID- 8407653 TI - Tenderness of beef loin steaks as influenced by marbling level, removal of subcutaneous fat, and cooking method. AB - Loins from 12 carcasses of Small minus marbling and loins from 12 carcasses of Slight minus marbling were selected. Steaks from these loins were either completely trimmed of external fat or trimmed to .6 cm of external fat. Steaks were cooked to 67 degrees C either by a rapid, high-temperature broiler-grill or by a slower, lower temperature electric broiler. Muscle on the dorsal side of steaks (where fat trimming was performed) was used for shear force measurements (five separate core locations), and the ventral portion was used for sensory evaluation of tenderness. Sensory scores for tenderness were affected (P < .05) by various first-order interactions, although differences were minimal. Steaks of Slight minus marbling cooked by broiler-grilling had reduced tenderness scores. Only when steaks were cooked by broiler-grilling did complete fat removal produce cores that appeared more well-done. Broiler-grilling produced more well-done cores than did electrical broiling. Correlations between shear force expressed as peak load and tenderness scores were highest in cores that were more similar in degree of doneness to sensory evaluation samples. Although broiler-grilling produced considerable within-steak variation in degree of doneness, some of the highest correlations between shear core degree of doneness and peak load and between peak load and sensory tenderness were obtained with this broiling procedure. This would imply that rapid broiling is important in creating uniform tenderness in loin steaks of reduced fat, but further modifications in rapid broiling cookery are necessary to improve uniformity of cooked meat color. PMID- 8407654 TI - Fermentation of raw poultry byproducts for animal nutrition. AB - In this study, the fermentation of raw, inedible poultry byproducts mixed with sugarbeet pulp and dextrose and inoculated with Lactobacillus plantarum and(or) Enteroccocus faecium resulted in a drop of pH in the byproducts to approximately 4.0 to 4.5 within 48 h. To keep the fermented product stable for a period of 21 d, the addition of > or = 3% (wt/wt) of a fermentable carbohydrate was necessary. With a high inoculation level of approximately 10(8) to 10(9) L. plantarum per gram, or with acidification of the initial mixture with .4% lactic acid, the number of Enterobacteriaceae decreased faster than with inoculation at 10(5) L. plantarum per gram, or without initial acidification. After 21 d of fermentation, a high level of enzymatic breakdown of proteins and amino acids was observed: the nonprotein N level increased from 5% to between 15 and 40% of total N and the volatile N level increased from 1% to between 3 and 11% of total N. An increase in histamine, cadaverine, and putrescine was also observed. Despite the technological measures taken, such as the application of a high inoculum of starter culture and initial acidification with .4% lactic acid, this amino acid breakdown could not be reduced to an acceptable level. These results suggest that, because of biochemical deterioration, fermentation alone is not a useful method of preservation of raw poultry byproducts. PMID- 8407655 TI - The feeding of raw, fermented poultry byproducts: using mink as a model. AB - In this study, the safety of fermentation as a method of preservation of raw animal byproducts used for animal nutrition was tested. Two feeding trials with mink, as a model for nonruminant animals, were carried out. In the first trial mink were given a fermented diet composed of raw poultry and fish byproducts supplemented with cereals, glucose, lactic acid, premix, and starter culture (Lactobacillus plantarum and Enterococcus faecium). These mink failed to deliver kits, and 7 of the 30 females in the test group died. At autopsy no specific cause of death could be diagnosed, although all the dead mink showed symptoms of cachexia. In a second trial, a group of mink kits, during the growth period, was given a diet composed of fermented poultry byproducts, just before feeding mixed with raw fish. The weight gain of the mink in the test group decreased statistically compared with that of the control group, mainly for the male members of the group. From the end of October until the beginning of November, during pelt priming, some mink showed symptoms of severe weight loss. It is suggested that the measured increase of amino acid breakdown, and(or) the acidic pH of the fermented diet, caused these unfavorable results. To examine the effect of the fermented diet on the gut flora, fecal samples were analyzed. The fermented diet changed the composition of the gut flora significantly. In the group that received the fermented diet the number of lactobacilli and the mesophilic aerobic count increased and the number of Enterobacteriaceae and enterococci decreased compared with the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407656 TI - Proteolysis and amino acid breakdown of heated and irradiated poultry byproducts and muscle tissue. AB - As a result of intensification and centralization of poultry slaughtering, the amount of slaughter byproducts produced at a single location is increasing. These byproducts are rich in protein, fat, and vitamins and, therefore, constitute a potentially useful raw material for use as animal feed. To maintain the nutritive value of these byproducts they should be processed to minimize or eliminate degenerative changes that reduce the feed value of the product. In this paper amino acid breakdown in slaughter-fresh poultry viscera, heads, and breast meat is studied as a model. Initial amino acid breakdown in viscera was observed (also when bacterial growth was excluded by gamma-irradiation), which resulted in high levels of total volatile N and cadaverine. Putrescine was produced only in viscera after bacterial proliferation. In heads and breast meat, no production of metabolites of amino acid degradation was observed as a result of initial enzymatic activity. It is concluded that during preservation of poultry byproducts not only bacterial proliferation, but also enzymatic breakdown of amino acids, must be prevented. PMID- 8407657 TI - Interrelationships between dietary protein level, energy intake, and nitrogen retention in pregnant gilts. AB - Two experiments were conducted to study the effects of dietary protein levels and DE intake on N retention in pregnant gilts. Thirty-two gilts were used in Exp. 1 to investigate the response to eight levels of dietary CP ranging from 50 to 235 g/kg (3.3 to 14.5 lysine/kg). Gilts were given 1,400 g of feed daily throughout pregnancy; diets contained similar balances of amino acids and similar amounts of DE (3.60 to 3.63 Mcal/kg). Thirty gilts in Exp. 2 were allocated during pregnancy to six levels of feeding ranging from 1.1 to 3.1 kg/d. The common diet given to gilts contained 3.49 Mcal of DE/kg, 155 g of CP/kg, and 10.7 g of lysine/kg and was considered adequate in protein. Nitrogen balance trials were conducted during early, mid-, and late pregnancy and collection periods of 5 d duration commenced on d 30, 58, and 86 in Exp. 1 and d 30, 58, and 93 in Exp. 2. The average live weights of pigs on all treatments within each collection period were similar and were 112.5, 123.3, and 136.6 kg and 120.7, 136.3, and 158.3 kg in Exp. 1 and 2, respectively. At each stage of pregnancy increments of dietary protein increased N retention up to an inflection point, after which N retention remained at a constant level. The maximum rates of N retention, 10.0, 12.1, and 16.5 g/d during early, mid-, and late pregnancy, occurred at 142, 133, and 162 g of CP/kg, respectively; the corresponding dietary lysine:DE values were 2.4, 2.3, and 2.7 g of lysine/Mcal of DE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407658 TI - The response of first-litter sows to dietary protein level during lactation. AB - Thirty-six sows were used to study responses of milk production, milk composition, and N balance to six concentrations of dietary CP ranging from 63 to 238 g/kg (4.4 to 15.1 g of lysine/kg) during the first lactation. Sows, on average, were 149.3 kg live weight and had 26.1 mm of back fat at P1 (which is 45 mm from the midline at the level of the last rib) immediately after parturition. During lactation, all sows suckled nine pigs each and were offered up to 4,000 g of feed daily; diets contained similar balances of amino acids and similar amounts of DE (3.56 to 3.63 Mcal/kg). Nitrogen balance trials were conducted during early and late lactation and 5-d collection periods commenced on d 10 and 24 of lactation, respectively. During both periods of lactation, there were significant positive linear relationships between the level of dietary protein and milk yield and contents of fat and total solids in milk. Milk yield increased from 7.79 to 9.91 kg/d and from 7.02 to 8.90 kg/d, whereas total solids in milk increased from 199 to 225 g/kg and from 202 to 228 g/kg during early and late lactation, respectively, in response to increasing level of dietary protein from 63 to 238 g of CP/kg. A two-phase linear regression model used to describe the relationship between N balance and dietary CP level established that sows required a diet containing > or = 202 g of CP/kg or 12.8 g of lysine/kg to maximize N balances during both stages of lactation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407659 TI - Effects of vitamin E and selenium on immune responses of peripheral blood, colostrum, and milk leukocytes of sows. AB - This study was designed to assess how dietary vitamin E (E) and (or) selenium (Se) concentrations affect immune responses of gestating and peripartum sows. Multiparous sows (24), assigned to one of four groups at breeding, were fed ensiled, shelled corn-soybean meal-based diets without supplemental E or Se (-E Se), with .3 mg of Se/kg (-E+Se), with 60 IU of E/kg (+E-Se), or with both supplemental E and Se (+E+Se) during gestation and to d 4 of lactation. Blood was obtained on 0, 30, 60, and 90 d of gestation and at parturition for serum E and Se assays. Lymphocytes and polymorphonuclear cells (PMN) were isolated from the blood, colostrum, and 4-d milk samples for immune studies. Compared with the control (+E+Se) diet, the -E-Se diet reduced (P < .05) the serum tocopherol and Se concentrations, the mitogenic responses of lymphocytes of peripheral blood (PBL) and colostrum (CL), the phagocytic activity of blood and colostral PMN, and the microbicidal activity of blood, colostral, and milk PMN. The -E+Se diet reduced (P < .05) the serum tocopherol concentrations, the mitogenic responses of PBL and CL, and the phagocytic activity of PBL. The +E-Se diet reduced (P < .05) serum Se concentrations and the phagocytic activity of PMN. The data indicated that E restriction depressed PBL and PMN immune functions, whereas Se restriction depressed mainly PMN function. PMID- 8407660 TI - Absorption balances and kinetics of nutrients and bacterial metabolites in conscious pigs after intake of maltose- or maltitol-rich diets. AB - Because sorbitol is poorly absorbed in the small intestine, it may be the origin of large amounts of residues reaching the large intestine and may be substrate for microbial activity. An experiment was conducted to study the quantitative appearance in the portal blood of nutrients and metabolites derived from enzymatic hydrolysis and microbial fermentation in the fore- and hindgut. Five Large White, castrated male pigs (mean BW of 61.2 +/- 1.7 kg) were fitted under anesthesia with an electromagnetic flow probe around the portal vein and with permanent cannulas in the portal vein and the carotid artery. From 10 d before surgery, they were accustomed to one of the two semisynthetic, well-balanced diets, containing a high level (53% of DM) of either a maltose-rich glucose syrup (SNat) or of a maltitol-rich hydrogenated glucose syrup (SHyd). Eight days after surgery and after an 18-h fast, each animal was given a last meal (800 g) of the diet to which it was formerly accustomed. For 12 h after this meal, blood samples were taken at 30- to 60-min intervals for glucose, sorbitol, amino N, VFA, D- and L-lactic acids, insulin, and glucagon determinations, and portal blood flow was continuously recorded. The absorption coefficients (amounts appearing for 12 h in the portal blood: amounts ingested, percentage) of glucose and of amino N were not significantly different between the two diets. The amount of sorbitol that appeared within 12 h in the portal blood after SHyd intake was 44 g (25% intake). The amount of VFA that appeared in the portal blood within 12 h was 2.7 times larger (P < .05) after intake of the maltitol-rich diet (SHyd:808 mmol) than after intake of the maltose-rich diet (300 mmol). This difference was due to an increase in absorbed amounts of propionate (SHyd 402 vs SNat 56 mmol, P < .05), butyrate (SHyd 63 vs SNat 17 mmol, P < .01), isovalerate (SHyd 17 vs SNat 5 mmol, P < .05), and acetate (SHyd 298 vs SNat 219 mmol, P < .13). There were no significant changes in insulin and glucagon production. Intake of the maltitol rich diet resulted in less available energy (82.0%) than did intake of the maltose-rich diet (92.6%). PMID- 8407661 TI - Characterization of toxicosis in sheep dosed with blossoms of sacahuiste (Nolina microcarpa). AB - To characterize more fully sacahuiste (Nolina microcarpa Watson) toxicosis in sheep and to evaluate benefits of supplemental Zn, sheep were dosed intraruminally with sacahuiste blossoms. In Trial 1, eight fine-wool sheep (47 +/ 2 kg BW) were fed alfalfa hay at 1% of BW daily and dosed intraruminally with inflorescences amounting to 1% of BW daily, in three portions per day, for 10 d. Four sheep were dosed intraruminally with aqueous ZnSO4 (30 mg of Zn/kg BW) daily for 3 d before initial sacahuiste dosing and on alternate days thereafter, and four sheep were untreated with Zn. Toxicosis was evident within 24 h after initial sacahuiste dosage, involving inappetence, depression, hypokalemia, hypophosphatemia, hyperbilirubinemia, and elevated serum enzymes (alkaline phosphatase, creatine kinase, lactate dehydrogenase, aspartate aminotransferase, and gamma-glutamyl transpeptidase). One sheep (untreated with Zn) died on d 3. Aqueous ZnSO4 alleviated inappetence and suppressed elevation of serum urea N and creatinine but did not suppress other changes in serum clinical profiles. In Trial 2, sacahuiste inflorescences were ruminally dosed into 12 fine-wool wethers (29 +/- 2 kg BW) in amounts equalling 0, .25, .50, .75% of BW per day, and chopped alfalfa hay was provided at 1.75% of BW per day for 14 d. Sacahuiste inflorescenses dosed at .75% of BW elicited severe toxicosis within 24 h, and dosage at .50 or .25% of BW per day increased (P = .12) serum bilirubin. Ruminal fluid pH, mean particle retention time, and particulate passage rate were not affected (P > .10) by sacahuiste, but ruminal fluid passage rate increased 1.6 fold (P < .10) and ruminal fluid volume decreased by 60% (P < .10) in sheep given inflorescenses amounting to .50% of BW daily. Sacahuiste inflorescenses dosed intraruminally at .75% of BW per day elicited ruminal impaction with severe hepatotoxicosis, and dosages amounting to .50% or .25% of BW per day caused similar trends. PMID- 8407662 TI - Cholecystokinin octapeptide immunization: effect on growth of barrows and gilts. AB - A study was conducted to validate the previously reported growth response to cholecystokinin octapeptide (CCK-8) immunization in barrows and was extended to include gilts. Group-penned barrows and gilts were used to represent conditions in the swine industry. Thirty-two animals, 19 barrows and 13 gilts, were randomly assigned by sex to four pens and two treatments. The control groups were immunized with human serum globulin (hSG). The treated groups (CCK) were immunized with the C-terminal octapeptide of cholecystokinin conjugated to human serum globulin. Specific binding of CCK-8 was confirmed at 29 d after the primary inoculation. Antisera titers were highly variable throughout. The mean titer reached a peak on d 57 and then declined. Body weight gains during the last 49 d, the period during which titers were expressed, were compared by ANOVA. The treatment effect on gain was significant (P = .018); the sex effect approached significance (P = .071); the treatment x sex interaction effect was not significant (P = .82). Least squares mean gain of the CCK group was 8.4% greater than of the hSG group, 41.4 vs 38.2 kg, respectively. A significant linear regression coefficient for gain vs antisera titer was obtained for barrows (P = .03; r2 = .44) but not for gilts. Several carcass variables showed trends similar to that of BW gain, but the treatment effects were less robust (P < .05 to .10). These results generally confirm the findings of the previous study; CCK-8 immunization stimulated growth of barrows by 7.5% in the present and by 10.8% in the previous study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407663 TI - Effects of cumulus cells on culture of bovine embryos derived from oocytes matured and fertilized in vitro. AB - Bovine zygotes derived from oocytes matured and fertilized in vitro were cultured in vitro. In Exp. 1, zygotes were vortexed to remove cumulus cells and then cultured in oviduct epithelial cell-conditioned TCM-199 medium with or without added cumulus cells. Embryos cultured without cumulus cells developed to blastocysts significantly less often than those cultured with added cumulus cells, 3 vs 38% of cleaved ova (P < .05). Co-culture of embryos with cumulus cells during the first 48 h only, or for the duration of the culture period, resulted in development rates similar to those of controls that were not vortexed. In Exp. 2, denuded zygotes were centrifuged (15,850 x g, 3 min) at room temperature to facilitate visualization of pronuclei. Centrifuged zygotes were either placed directly into conditioned medium for culture (with or without added cumulus cells) or DNA was first microinjected into their cytoplasm or a pronucleus. All microinjected embryos were co-cultured with added cumulus cells. Centrifugation and microinjection did not reduce viability significantly when embryos were co-cultured with cumulus cells. Development was reduced significantly when embryos were cultured without cumulus cells. Thus, when oviduct epithelial cell-conditioned medium was used, cumulus cell co-culture was beneficial during the early cleavage stages of in vitro-derived zygotes, including those that received DNA microinjection. PMID- 8407664 TI - Effect of synchronization of energy and nitrogen supply on ruminal characteristics and microbial growth. AB - The effect of energy and N synchronization in the rumen on microbial growth was investigated. The same daily amount of readily available energy and N sources (19 g of ruminally degradable N/kg of fermentable OM) was supplied intraruminally to sheep, according to different patterns, namely both energy and N as 12-hourly pulse-doses (fast synchronized supply), energy as 12-hourly pulse-doses and N as a continuous infusion (unsynchronized supply), energy as a continuous infusion and N as 12-hourly pulse doses (unsynchronized supply), and both energy and N as continuous infusions (slow synchronized supply). The study was done near maintenance (Exp. 1) and at a higher level of nutrition (Exp. 2). The degree of energy and N synchronization affected neither microbial flow nor efficiency of growth (P > .2 for energy x N interaction). Continuous infusion of energy resulted in a 17% (P < .05) and 14% (P = .18) higher efficiency of microbial growth than did pulse dosing in Exp. 1 and 2, respectively. This coincided with lower (P < .01) ruminal pH and higher (P < .05) ruminal lactic acid concentration for energy pulse-dose treatments. The results suggest that merely improving the degree of synchronization between energy and N release rates in the rumen does not increase microbial yield. Dietary manipulation, rather, should be aimed at first obtaining the most even ruminal energy supply pattern, and then at providing the appropriate amount of ruminally available N. Thereafter some further advantage may be gained in also ensuring a more even N supply pattern, particularly avoiding too rapid a ruminal N release. PMID- 8407665 TI - Net flux of metabolites across the ruminal wall of sheep fed twice a day with orchardgrass hay. AB - Four Texel wethers (68 +/- 2.5 kg BW) fitted with catheters in the ruminal veins and a mesenteric artery, blood flow probes around ruminal arteries, and a ruminal cannula were used to determine meal-related variations and daily significance of net flux across the ruminal wall of urea and ammonia (NH3), VFA, D-beta hydroxybutyrate (D beta HOB), lactate, and glucose. Sheep were fed every 12 h with orchardgrass hay (430 g of DM/meal; 611 g of digestible OM/kg of DM and 23.6 g of N/kg of DM). Apart from lactate and glucose, the fluxes of studied metabolites were significantly affected by time after morning feeding. Maximum absorption of VFA and NH3 were observed at the end of the meal; however, 5 h after the meal VFA absorption was still high, whereas NH3 absorption had decreased to the prefeeding level. Net release of D beta HOB was greater during the 2 h after the meal than during the rest of the time. Urea net transfer decreased during the meal, and thereafter it increased to the 5th h after feeding, at which time it was twofold higher than at prefeeding. The difference in net flux across the ruminal wall of urea and NH3 was linearly correlated with NH3 concentration in the ruminal fluid. Daily urea and NH3 net transfer were 2.10 and 3.76 g of N/d, respectively. The VFA net appearance in the ruminal veins was 1.167, .226, and .014 mol/d for acetate, propionate, and butyrate, respectively. Daily net release of D beta HOB, lactate, and glucose by the rumen wall was .153, .093, and -.012 mol/d, respectively. PMID- 8407666 TI - Stress induction affects copper and zinc balance in calves fed organic and inorganic copper and zinc sources. AB - This study determined whether Cu and Zn balance was affected by feeding either Zn methionine (ZnMet) + Cu lysine (CuLys) or Zn sulfate (ZnSO4) + Cu sulfate (CuSO4) before and after stressing calves. Eight Charolais crossbred steer calves weighting 167 +/- 5 kg were randomly assigned to two treatments in a crossover experimental design. The millet hay and soybean meal diet when supplemented with the inorganic salts provided 9.2 ppm of Cu and 36.6 ppm of Zn or when fortified with the metal complexes contained 10.5 ppm of Cu and 36.6 ppm of Zn. Gentled calves were fed their respective diets for 28 d before an 18-d mineral balance trial was conducted. Collection consisted of five periods: 1) a 5-d baseline period, 2) 3 d of no Cu and Zn supplement, 3) 3 d of stress consisting of feed and water restriction and ACTH (80 IU) injections i.m. every 8 h, 4) 3 d of refeeding with no Cu and Zn supplement, and 5) 4 d of Cu and Zn repletion. Calves fed CuLys had 53% greater apparent Cu absorption and increased Cu retention (P < .05) during repletion compared with calves fed CuSO4. The 18-d mean retention of Cu from CuLys was greater (P < .05) than that from CuSO4. No differences (P > .05) in apparent absorption or retention of Zn were found between Zn sources, although during the 18-d trial mean retention was 58% higher when ZnMet was fed. Urinary Cu and Zn excretion decreased (P < .01) during stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407667 TI - Influence of supplementation on behavior of grazing cattle. AB - Dormant grasses are usually deficient in nutrients needed to meet cattle requirements. Research results indicate that grazing activities are sensitive to some environmental variables and vegetative (sward) characteristics; however, minimal information is available on the influence of supplementation regimens on cattle grazing activities. Supplementation studies reviewed had forage N levels that ranged from .9 to 1.2% and forage intakes that ranged from 12.3 to 31.0 g of OM/kg of BW. Protein supplementation affected (P < .05) time spent grazing: unsupplemented (NS) cattle grazed approximately 1.5 h/d more than did supplemented (S) cattle. Type of supplemental protein and time of daily feeding did not affect (P > .10) this response. Different types and timing of starch based supplements produced variable results; however, increasing the level of supplemental starch decreased (P < .05) daily grazing time. Protein supplementation increased (P < .05) harvesting efficiency (HE; grams of forage intake.kilogram of BW-1.minute spent grazing-1); however, high-starch supplements either did not alter (P > .10) or decreased (P < .05) HE compared with NS. Progressive defoliation can influence grazing behavior in both sheep and cattle grazing actively growing forage; however, evaluation of vegetative characteristics of dormant forage and the corresponding effect on grazing behavior are not known. The effects of various grazing management strategies on cattle behavior are inconclusive and deserve additional attention. Methods and (or) management practices that modify behaviors to control feed intake, improve efficiency, or reduce stress could be major contributions to the livestock industry. PMID- 8407668 TI - Naturally occurring toxins in feedstuffs: Center for Veterinary Medicine Perspective. AB - The objectives of this review are to provide 1) information on the FDA Feed Contaminants Program, 2) the legal history of aflatoxins and their current action levels, 3) a report on the levels of aflatoxins, fumonisins, vomitoxin, ochratoxin A, and zearalenone in domestic and import surveillance samples of feed during fiscal years 1989 through 1992, and 4) information on naturally occurring toxins encountered recently by the Center for Veterinary Medicine. Ten of 644 (1.6%) domestic corn samples and 7 of 106 (6.6%) domestic cottonseed samples contained aflatoxins at levels > 300 ppb. The mean fumonisin level in the 1990 survey of 85 corn screening samples was 12.1 ppm, and the values ranged from 2.6 to 32 ppm. The mean vomitoxin levels in the 1991 survey of 207 winter wheat samples and 206 spring wheat samples was 2.4 and .9 ppm, respectively. Ochratoxin A was not detected in 168 samples. Zearalenone was detected at levels > .15 ppm in only 1 of 161 samples. Cottonseed containing 13,000 ppm gossypol was recently implicated in the deaths of dairy cows. Crambe meal and canola meal are sanctioned for use in feed with certain restrictions, including the levels of glucosinolates. The FDA is continuing its surveillance and will strive to provide guidance on the increasing number of naturally occurring toxins. PMID- 8407669 TI - Analysis of naturally occurring mycotoxins in feedstuffs and food. AB - Aflatoxins, zearalenone, deoxynivalenol, fumonisins, and their respective metabolites require specific procedures for their determination because of their diverse chemistry and occurrence in complex matrices of feedstuffs and foods. Major sources of error in the analysis of these mycotoxins arise from inadequate sampling and inefficient extraction and cleanup procedures. The determinative step in the assay for each of these toxins is sensitive to levels below those that are considered detrimental to humans and animals. Aflatoxins can be determined in grains and animal fluids and tissues by TLC, HPLC, gas chromatography-mass spectrometry (GC-MS), and ELISA procedures. Zearalenone, an estrogenic mycotoxin, can readily be determined in cereal grains and foods by HPLC (50 ng/g) and by TLC (300 ng/g). No incurred levels of zearalenone or its metabolites have been detected in animal tissues destined for human consumption. Deoxynivalenol can be determined in wheat and corn at 300 ng/g by a rapid TLC procedure and at 325 ng/g by a GC method. Although not tested collaboratively, an HPLC procedure and an ELISA screening procedure are capable of detecting deoxynivalenol at low (nanograms/gram) levels in feedstuffs and foods. The recently characterized fumonisins can be detected by TLC, HPLC, and GC-MS at levels below those now considered harmful. Thin-layer chromatography and HPLC (with fluorescence detection of derivatives) procedures can detect fumonisins at approximately 100 ng/g; GC-MS is required for detection at lower levels. PMID- 8407670 TI - Effects of low concentrations of ampicillin in feed on the intestinal Escherichia coli of chicks. AB - Ampicillin in low concentrations (1.7 and 5 g t-1) was incorporated in the feed given to 1-d-old chicks for 2 weeks. This was sufficient to select, in the intestinal contents, resistant Escherichia coli strains for which the minimum inhibitory concentration of ampicillin was > 100 micrograms ml-1. Different clones of E. coli were identified by their biotype, pattern of resistance to antibacterial agents and plasmid profile (designated P-P types). The experiment was repeated twice and the average proportion of ampicillin-resistant P-P types among 72 isolates of E. coli from chicks given feed containing 0, 1.7 and 5 g ampicillin t-1 were 10%, 31% and 46% respectively. PMID- 8407671 TI - Antimicrobial activity of lactic acid bacteria isolated from sour doughs: purification and characterization of bavaricin A, a bacteriocin produced by Lactobacillus bavaricus MI401. AB - Three hundred and thirty-five lactic acid bacteria were isolated from sour doughs and screened for antagonistic activity. Of these 145 showed activity against one or several of the indicator strains used in the screening. The antimicrobial activity of 18 isolates were due to a proteinaceous compound. These 18 isolates belonged to three different Lactobacillus species: Lactobacillus bavaricus, Lactobacillus curvatus and Lactobacillus plantarum. The spectrum of antimicrobial activity for the three species suggested that the inhibitory components were different. The inhibitory compound from Lact. bavaricus MI401 was chosen for further study. The proteinaceous nature, antimicrobial activity against closely related species, heat resistance and sensitivity to alkaline treatment strongly indicated that this substance was a bacteriocin, which we designated bavaricin A. The bacteriocin was purified to homogeneity by ammonium sulphate precipitation, ion exchange, hydrophobic interaction and reverse-phase chromatography. The purification resulted in 193,000-fold increase in specific activity. SDS-PAGE of bavaricin A showed a molecular weight of 3500-4000 Da. By amino acid sequencing 41 amino acids were determined. Bavaricin A had a bactericidal mode of action and inhibited nine out of 10 Listeria monocytogenes. Lactobacillus bavaricus MI401 produced bavaricin A at temperatures from 4 degrees C to 30 degrees C. The production of active bavaracin A was inhibited at increasing sodium chloride concentration. In the presence of 3% sodium chloride at 4 degrees C no active bavaricin A could be detected. Nitrite (100 ppm) did not affect the production of active bavaricin A. PMID- 8407672 TI - Effect of sub-inhibitory concentrations of antibiotics on the virulence of Staphylococcus aureus. AB - The production of virulence factors by various bacteria can be influenced by sub inhibitory concentrations of antibiotics. The effect of six antibiotics on the production of representative extracellular enzymes and toxins produced by Staphylococcus aureus was investigated. The production of the virulence determinants coagulase, protein A, alpha and delta haemolysin was monitored in the presence of ciprofloxacin, enoxacin, chloramphenicol, gentamicin, tetracycline and methicillin. The protein synthesis inhibitors reduced the production of coagulase and protein A, and almost completely inhibited the production of the haemolysins. Haemolysin production was also reduced by ciprofloxacin and enoxacin, but these antibiotics had little effect on the production of coagulase and protein A. Methicillin stimulated the production of alpha and delta haemolysins but had no effect on the production of coagulase and protein A. PMID- 8407673 TI - The effect of tetronasin and monensin on fermentation, microbial numbers and the development of ionophore-resistant bacteria in the rumen. AB - The Gram-negative rumen bacteria Fibrobacter succinogenes S85, Prevotella ruminicola M384 and Veillonella parvula L59 were grown in media containing successively increasing concentrations of the ionophores, monensin and tetronasin. All three species became more resistant to the ionophore with which they were grown. Increased resistance to one ionophore caused increased resistance to the other, and cross-resistance to another ionophore--lasalocid- and an antibiotic--avoparcin. Recovery of tetronasin-resistant bacteria from the rumen of monensin-fed sheep increased and vice versa, indicating that similar cross-resistance occurred in vivo. PMID- 8407674 TI - Bacteriocinogenic activity of lactobacilli from fermented sausages. AB - During the screening of the inhibitory activity of 254 strains of lactobacilli isolated from fermented sausages at different times of ripening, 22% of the strains showed inhibition that was not related to acid or hydrogen peroxide, towards one or more indicator strains. Not all the strains were capable of secreting the inhibitory compound in the supernatant fluid. The characterization of the inhibitory compound from three strains showed that they were bacteriocins with a bactericidal mode of action and a molecular weight exceeding 10,000 Da. Lactobacillus plantarum CTC 305, CTC 306 and Lact. sake CTC 372 inhibited Listeria monocytogenes. Lactobacillus sake CTC 372 was cured of two plasmids of 84.8 kbp and 41.3 kbp, losing the production and the immunity of a bacteriocin as well as the ability to ferment lactose. PMID- 8407675 TI - A note on the primary structure and expression of an Erwinia carotovora polygalacturonase-encoding gene (peh1) in Escherichia coli and Saccharomyces cerevisiae. AB - A 1209-base pair (bp) DNA fragment containing the endopolygalacturonase-encoding gene (peh1) from Erwinia carotovora subsp. carotovora was amplified by the polymerase chain reaction (PCR) technique and expressed in Escherichia coli. The nucleotide sequence of the PCR product was determined and found to be highly homologous to the primary structures of other polygalacturonase-encoding genes. The peh1 DNA fragment encoding the mature polygalacturonase was inserted between two different yeast expression-secretion cassettes and a yeast gene terminator, generating recombinant yeast-integrating shuttle plasmids pAMS10 and pAMS11. These YIp5-derived plasmids were transformed and stably integrated into the genome of a laboratory strain of Saccharomyces cerevisiae. Transcription initiation signals present in these expression-secretion cassettes were derived from the yeast alcohol dehydrogenase (ADC1P) or mating pheromone alpha-factor (MF alpha 1P) gene promoters. The transcription termination signals were derived from the yeast tryptophan synthase gene terminator (TRP5T). Secretion of polygalacturonase was directed by the signal sequence of the yeast mating pheromone alpha-factor (MF alpha 1S). Northern blot analysis revealed the presence of peh1 mRNA in the yeast transformants and a polypectate agarose test was used to monitor polygalacturonase production. PMID- 8407676 TI - A comparison of three salmonella antigen-capture ELISAs and culture for veterinary diagnostic specimens. AB - Three sandwich-ELISAs, two of which are commercially available (Tecra and Locate), and one developed at the Veterinary Sciences Division, Stormont and a 3 step culture protocol, were compared for the detection of salmonella in 1000 animal specimens. Eight hundred and fifty of these were new submissions and the remainder were frozen portions from specimens previously shown to contain salmonellas by culture. The incidence of ELISA false-negative and false-positive results was highest for the Stormont and Locate kits respectively although the differences in sensitivity and specificity between the three ELISAs was not statistically significant. On 16 occasions all ELISA methods indicated the presence of salmonellas when none were isolated by initial culture, eight of these specimens contained salmonellas when reinvestigated by culture. PMID- 8407677 TI - Isolation and characterization of carboxylesterase E3 from Salmonella enterica. AB - Three esterases (Est-) hydrolysing alpha-naphthyl acetate: Est-E1, Est-E3 and Est E4 produced by Salmonella enterica serovar Typhimurium, strain LT2 were separated by DEAE chromatography and gel filtration. Est-E3, the major component of this set of enzymes, clearly differed from the two other esterases by its apparent molecular weight, titration curve, substrate specificity and inactivation. Immunoglobulins raised against Est-E3 completely neutralized the activity of Est E3 but did not react with Est-E1 or Est-E4; it showed no cross reaction with carboxylesterase B of Escherichia coli or with carboxylesterases from other enterobacteria. Est-E3 showed electrophoretic variants which were biochemically and immunologically detected in the seven subspecies of the genus Salmonella. These findings suggest that variants of Est-E3 are the products of very closely related loci originating from a common ancestral gene. The esterase could be a phylogenetic marker of the genus and a suitable molecular tool for taxonomy and epidemiology. PMID- 8407678 TI - Lactic acid bacteria and human clinical infection. PMID- 8407679 TI - Effects of invented spelling and direct instruction on spelling performance of second-grade boys. AB - Four second-grade boys, 2 rated by their classroom teacher as below average and 2 as above average in basic language skills, participated in a 16-week spelling investigation. The participants alternately received, in counterbalanced order, 5 weeks of an invented spelling approach that incorporated 15-min creative writing periods and 5 weeks of direct instruction that involved 15-min periods of guided practice on spelling word lists. At the end of 10 weeks, each condition was replicated for 3 additional weeks. Although direct instruction resulted in more targeted words spelled correctly, invented spelling resulted in more nontargeted words spelled correctly, higher preference ratings by children, and higher teacher ratings of the quality of 3 of the children's writing samples. PMID- 8407680 TI - Descriptive and experimental analyses of variables maintaining self-injurious behavior. AB - Independent descriptive (correlational) and functional (experimental) analyses were conducted to determine the extent to which the two methods would yield data supporting similar conclusions about variables maintaining the self-injurious behavior (SIB) of 6 subjects. For the descriptive analyses, subjects were observed in their residences and at training sites at various times each day while observers recorded naturally occurring sequences of specified subject and staff behaviors. The subjects also participated in a day program for the assessment and treatment of SIB, in which they were exposed to functional analyses that manipulated potential maintaining variables in multielement designs. Both sets of data were analyzed via conditional probabilities to identify relevant antecedent and consequent events for subjects' SIB. Using outcomes of the experimental analysis as the standard for comparison, results indicated that the descriptive analysis was useful in identifying the extent to which SIB was related to social versus nonsocial contingencies, but was limited in its ability to distinguish between positive and negative reinforcement (i.e., attention versus escape). PMID- 8407681 TI - Effects of setting events on the problem behavior of students with severe disabilities. AB - Two studies analyzed the effects of preceding setting events on the problem behavior of students with severe disabilities. Using ABAB withdrawal designs, the occurrence versus nonoccurrence of preceding setting events was analyzed in relation to the frequency of problem behavior. Data were collected throughout a student's school day, with interventions focusing upon the elimination of setting events that occurred before school. The results indicate that (a) the occurrence of preceding setting events was related to higher frequencies of problem behavior and (b) interventions designed to eliminate preceding setting events were consistently associated with low frequencies of problem behavior. PMID- 8407682 TI - Differential effects of self-monitoring attention, accuracy, and productivity. AB - Effects of self-monitoring on-task behavior, academic productivity, and academic accuracy were assessed with 6 elementary-school students with learning disabilities in their general education classroom using a mathematics task. Following baseline, the three self-monitoring conditions were introduced using a multiple schedule design during independent practice sessions. Although all three interventions yielded improvements in either arithmetic productivity, accuracy, or on-task behavior, self-monitoring academic productivity or accuracy was generally superior. Differential results were obtained across age groups: fourth graders' mathematics performance improved most when self-monitoring productivity, whereas sixth graders' performance improved most when self-monitoring accuracy. PMID- 8407683 TI - Extinction of self-injurious escape behavior with and without instructional fading. AB - Three individuals with developmental disabilities participated in a study of the treatment of self-injurious behavior (SIB) maintained by negative reinforcement (escape from educational tasks). Treatment was implemented in a multiple baseline design across subjects, in which two treatments were compared in a multielement format. Both treatment conditions included an escape-extinction component in which SIB no longer produced escape. One of the conditions also included a fading component in which the frequency of instructions was initially reduced to zero and then was gradually faded back in across sessions until the instructional rate matched that of the original baseline. Results indicated that extinction alone reduced SIB to the end-of-treatment criterion in fewer sessions than did extinction plus fading for all 3 subjects. For 2 of the 3 subjects, however, there was an initial increase in the frequency of SIB at the outset of treatment with extinction (an extinction burst) that was not observed when extinction was combined with the fading component. PMID- 8407684 TI - The failure of feedback on alcohol impairment to reduce impaired driving. AB - We examined the effects of rules to govern drinking, individual feedback on blood alcohol concentration (BAC), and public posting of group data on impaired driving on the incidence of impaired driving. Level of impairment was determined from breath samples taken from tavern patrons. Following baseline, an intervention package consisting of (a) cards to guide patrons in pacing their drinking to stay under the legal limit, (b) individual feedback on BAC, and (c) posted group feedback on the percentage of patrons driving while impaired the preceding week was introduced in two taverns. Results indicated that the intervention package did not reduce the percentage of impaired drivers departing either tavern. The addition of a brief intensive police enforcement program directed at impaired driving produced a short-term reduction in impaired driving. PMID- 8407685 TI - Treatment of social behavior in autism through the modification of pivotal social skills. AB - We examined acquisition of individual social communicative behaviors and generalization across other social behaviors in 2 children with autism. The results of a multiple baseline design showed that the children's treated social behaviors improved rapidly and that there were generalized changes in untreated social behaviors. These improvements were accompanied by increases in subjective ratings of the overall appropriateness of the children's social interactions. The results suggest the possibility of identifying pivotal response classes of social communicative behavior that may facilitate the understanding of social behavior in autism as well as improve peer interactions, social integration, and social development. PMID- 8407686 TI - Improving dietary practices of elderly individuals: the power of prompting feedback, and social reinforcement. AB - Three intervention packages consisting of (a) enhanced prompts, feedback, and social reinforcement; (b) a lottery; and (c) serving as a confederate were added and removed in sequence as adjacent conditions in an extended withdrawal design to assess their effects on the dietary choices of elderly persons. Participants were 3 elderly residents of an independent living facility who were identified as making consistently poor dietary choices and who had medical conditions that necessitated changes in their eating habits. All 3 participants demonstrated a marked increase in healthy choices of food items in response to the package of enhanced prompts, feedback, and social reinforcement. No additional increase occurred with the introduction of the lottery and serving as a confederate. Food choice data indicated that most of these improvements could be attributed to healthier entree and dessert choices. Group data for all residents suggested small improvements in dietary practices during the three intervention conditions, with the largest proportion of the group's healthy choices occurring when the lottery was added to enhanced prompts, feedback, and social reinforcement. Food choice data indicated that most of these improvements could be attributed to healthier dessert choices alone. PMID- 8407687 TI - An evaluation of two methods for increasing self-initiated verbalizations in autistic children. AB - Three children with autism and mental retardation were treated for deficits in self-initiated speech. A novel treatment package employing visual cue fading was compared with a graduated time-delay procedure previously shown to be effective for increasing self-initiated language. Both treatments included training multiple self-initiated verbalizations using multiple therapists and settings. Both treatments were effective, with no differences in measures of acquisition of target phrases, maintenance of behavioral gains, acquisition with additional therapists and settings, and social validity. PMID- 8407688 TI - Effects of community and center-based settings on the alertness of persons with profound mental retardation. AB - We studied effects of different settings on the behavior of persons with profound mental retardation. Adaptive (alert) and nonadaptive behaviors were observed in two community settings and a center setting. Results of the descriptive assessment showed that participants engaged in a higher percentage of adaptive behaviors and a lower percentage of nonadaptive behaviors in the "high stimulation" community setting. The results are discussed in light of environmental setting events on persons with profound mental retardation. PMID- 8407689 TI - Efficacy and maintenance of an education program for a consumer cooperative. AB - We examined the effects of contingency management on participation in and maintenance of an education program by new members of a student housing cooperative. With credit and fine contingencies in place, the percentage of participants completing study guides was five times higher than without the contingencies. Members continued to implement the program for 9 years without researcher involvement. PMID- 8407690 TI - Effects of a multifaceted training procedure on the social behaviors of hearing impaired children with severe language disabilities: a replication. AB - The effectiveness of a training package on the social behaviors of 20 hearing impaired children with severe language disabilities was assessed. The package consisted of initial instruction in role-play situations, reinforcement of appropriate instances of the behaviors, and a correction procedure following inappropriate instances of the behaviors. Results showed that the training package was effective in increasing greeting, turn waiting, initiating interaction, and giving help and these effects were maintained over several weeks. PMID- 8407691 TI - Prostatitis--an increasing clinical problem for diagnosis and management. AB - Prostatitis remains a challenging condition. The clinical features are often nonspecific while the aetiology and pathogenesis can be diverse and includes inflammatory, obstructive, and/or chemical causes and may also be related to calculi. Four categories are recognized: acute bacterial prostatitis, chronic bacterial prostatitis, non-bacterial prostatitis and prostatodynia. The diagnosis of prostatitis was advanced substantially by the introduction of sequential sampling of urine aliquots following prostatic massage. Bacterial prostatitis is largely associated with the Enterobacteriaceae although Pseudomonas spp., enterococci and Staphylococcus aureus may also be isolated. In chronic bacterial prostatitis a variety of streptococci and anaerobic bacteria may be isolated. Treatment is difficult largely owing to the limited range of agents able to achieve therapeutic concentrations within prostatic fluid, which has a pH lower than that of plasma. Trimethroprim, co-trimoxazole and the tetracyclines have been widely used. The quinolones have recently been shown to diffuse readily into the prostate; ofloxacin and temafloxacin have produced the highest concentrations in prostatic fluid. Antibiotic treatment requires prolonged high dosage and careful monitoring to ensure that bacterial eradication has occurred. Other forms of management have included the judicious use of anti-inflammatory agents and analgesics. In some patients zinc sulphate has proved to be of symptomatic benefit. PMID- 8407692 TI - Treatment of chronic viral hepatitis. AB - A substantial number of anti-viral compounds have been evaluated for the treatment of patients with chronic viral hepatitis. A few of these compounds have now achieved clinical applicability. alpha-Interferon is the most widely studied and remains the main treatment for chronic hepatitis B and C. Unfortunately in both these conditions only a minority of patients respond to interferon therapy, although the response can be complete in some patients. Some parameters have been identified which assist in the selection of patients for treatment. Several other cytokines, including thymosin, have been evaluated for the treatment of chronic hepatitis B. There are a number of promising new nucleosides which may inhibit hepatitis B virus and their action is being studied. Relapse rates are unknown however with these compounds. Ribavirin, a guanosine analogue, is also efficacious in treating a proportion of patients with chronic hepatitis C and the drug may be useful in treating patients with cirrhosis or patients who have an auto-immune diathesis. PMID- 8407693 TI - The domiciliary management of genital tract infection. AB - Present treatment schedules for lower genital tract infections are far from ideal. Members of the 4-quinolone antibiotic class are highly active against Neisseria gonorrhoeae, and some of the newer agents are active against Chlamydia trachomatis. Clinical success has been reported with, for example, a single daily dose of ofloxacin for one week. New macrolides are active against C. trachomatis and N. gonorrhoeae. Azithromycin has been shown to be effective in a single oral dose. The role of newer agents in the therapy of bacterial vaginosis is largely unproven, but there are encouraging signs. The importance of attempting to make a firm diagnosis is emphasized. PMID- 8407694 TI - Antivirals for the treatment of herpesvirus infections. AB - Agents available to treat herpesvirus infections include idoxuridine, trifluridine, vidarabine and acyclovir for the topical treatment of herpetic eye infections; vidarabine and acyclovir for the systemic (intravenous) treatment of herpes encephalitis; acyclovir for the topical and systemic (oral) treatment of genital herpes; acyclovir for the systemic (intravenous, oral) treatment of HSV or varicella-zoster (VZV) infections in immunosuppressed patients; brivudin for the systemic (oral) treatment of HSV-1 or VZV infections in immunosuppressed patients; and ganciclovir and foscarnet for the systemic (intravenous) treatment of cytomegalovirus (CMV) retinitis in AIDS patients. Brivudin is also effective in the treatment of herpetic eye infections that no longer respond to idoxuridine, trifluridine, vidarabine or acyclovir; and foscarnet is effective in the treatment of infections with acyclovir-resistant, thymidine kinase-deficient (TK-) HSV or VZV mutants. Other antiviral agents considered for use in herpesvirus infections include brovavir, penciclovir (and its prodrug famciclovir), desciclovir (a prodrug of acyclovir), bishydroxymethylcyclobutylguanine (BHCG) and, in particular, 1-(3-hydroxy-2 phosphonylmethoxypropyl)cytosine (HPMPC). The latter is more active than either acyclovir or ganciclovir in the chemotherapy and prophylaxis of various HSV-1, HSV-2, TK- HSV, VZV or CMV infections in animal models. PMID- 8407695 TI - Current use of anti-HIV drugs in AIDS. AB - Since 1987 major advances have been made in our understanding of the pathogenesis of infection and the possible inhibition of the HIV virus. Various drugs targeted to the different steps of viral replication have been selected, but drugs such as soluble CD4 or dextran derivatives aimed to inhibit or interfere with the GP120 CD4 attachment step have shown little or no clinical benefit. Protease inhibitors or interferons acting at the post-transcriptional level are currently under phase I to II investigation. The only group of compounds clinically active belong to the nucleosides analogues that act as DNA chain terminators and by inhibiting viral reverse transcriptase. Zidovudine (AZT), didanosine (ddI) and dideoxycytidine (ddC) have been extensively studied, and used on a large clinical scale. Stavudine (D4T), deoxyfluorothymidine (FLT) and 3'thiacytidine (3TC) are entering phase I-II studies. Being the first nucleoside analogue discovered and used since early 1985, zidovudine remains the gold standard of anti-HIV therapy. Zidovudine is indicated at a dosage varying between 500 and 1000 mg in patients with AIDS and ARC, in asymptomatic patients with CD4 < 200/mm3 and in asymptomatic patients with CD4 between 200 and 500 cells/mm3 with a rapid decrease of CD4 cell count or a positive P24 circulating antigen. There is as yet no consensus concerning patients with more than 500 cell/mm3. ddI and ddC are currently indicated for patients intolerant to AZT or in those who have not responded (clinically or biologically) to AZT. Emergence of resistance to AZT has been reported in 30 to 80% of patients at various stages of the disease and after six to nine months of therapy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407696 TI - Aetiology and treatment of community-acquired pneumonia in adults: an historical perspective. AB - Community-acquired pneumonia is common. Most disease is mild but mortality among hospitalized patients is 5-20%. The most common aetiological pathogens are Streptococcus pneumoniae, Haemophilus influenzae, and the 'atypical' organisms, Mycoplasma pneumoniae, Legionella pneumophila and Chlamydia pneumoniae. Less common pathogens account for 10-30% of cases and the aetiology cannot be determined in one-third to one-half of cases. Classification by aetiology and initiation of specific antimicrobial therapy are difficult and treatment is often initiated empirically. Ampicillin (or amoxycillin) or erythromycin are inexpensive and effective for most patients, but their use in combination, the addition of a beta-lactamase inhibitor (e.g. amoxycillin/clavulanate) or the substitution of an expanded spectrum cephalosporin (e.g. cefuroxime) should be considered for patients with more serious illnesses or pathogens likely to be drug-resistant. Fluoroquinolones such as ciprofloxacin or ofloxacin would be acceptable if adequacy for treating pneumococcal infections were likely. New macrolides, such as azithromycin and clarithromycin, and new fluoroquinolones, such as temafloxacin and sparfloxacin, have theoretical advantages over previously available drugs, but superior efficacy has not yet been demonstrated satisfactorily. Pneumococcal resistance in various parts of the world is modifying traditional treatment. Currently, there is no drug of choice for the empirical treatment of community-acquired pneumonia. PMID- 8407697 TI - Diagnostic problems in lower respiratory tract infections. AB - The main problems of diagnosis in lower respiratory tract infection are the differentiation of infection from colonization or contamination, and the isolation of a reliable and true pathogen. The clinical findings and differentiation of patients into those with pneumonia or infective exacerbations of chronic bronchitis should provide a definitive early diagnosis. Expectorated sputum may be unreliable in pneumonia, because of contamination by oropharyngeal flora. Although blood cultures may be negative, they provide a precise diagnosis and should be obtained in all pneumonias admitted to hospital. Other more invasive procedures are transtracheal needle aspiration, fibrebronchoscopic techniques including protected specimen brush and bronchoalveolar lavage with quantitative culturing and cytological analysis, transthoracic needle aspiration, thoracoscopy--guided biopsy and open lung biopsy. Any invasive procedure in a severely ill patient should be carefully directed weighing the risks as well as the benefits, whilst taking the underlying disease and expected survival into consideration. PMID- 8407698 TI - Management of bacterial meningitis. AB - In developed countries the mortality from bacterial meningitis acquired outside the neonatal period is relatively low. In contrast, in developing countries it is often higher (20%-40%). In developed countries despite (and perhaps because of) the introduction of increasingly potent antimicrobials, the morbidity of bacterial meningitis has remained high. For example, up to 25% of patients with Haemophilus influenzae meningitis have some form of neurological deficit. Neisseria meningitidis is the major cause of bacterial meningitis in many areas of the world. A clone of Group A meningococcus has spread from China to cause the most recent major epidemic in Sub-Saharan Africa. Group B meningococcal infections causing sporadic meningitis are increasing in parts of Europe and South America. The mortality from meningococcal disease is greatest when there is a septicaemic component to the infection. Although antimicrobial chemotherapy is of major importance some adjuncts to therapy are beneficial. High dose corticosteroid therapy has been shown to decrease mortality in pneumococcal meningitis in an uncontrolled study and to speed recovery and decrease neurological sequelae in H. influenzae meningitis. Nevertheless to prevent infection would be of greater benefit. Prevention can be achieved by either chemoprophylaxis or immunoprophylaxis. Although safe and effective vaccines are available to prevent pneumococcal, H. influenzae (Hib) and Groups A and C meningococcal meningitis; apart from the protein conjugate Hib vaccine they are less effective in children under two years of age. There is no effective vaccine to protect against group B meningococcal meningitis. PMID- 8407699 TI - Helicobacter pylori infection. AB - The discovery of Helicobacter pylori is arguably the most significant advance made in gastroduodenal pathology this century. It is the most important cause of chronic gastritis, and almost certainly the major aetiological factor responsible for duodenal ulcer and probably for gastric ulcer as well. Evidence is accumulating which suggests that it may play an important role in the pathogenesis of gastric cancer. H. pylori is thought to be transmitted by the faecal-oral route or possibly oral-oral route, with iatrogenic transmission also reported. The prevalence of H. pylori infection increases with age, is commonest in developing countries, in certain ethnic minorities and those in lower socio economic and educational groups. The organism can be eradicated using combinations of antibiotics; when treatment is successful inflammatory changes resolve, duodenal ulcers heal and do not subsequently recur. PMID- 8407700 TI - Foreign body associated infection. AB - One of the major complications associated with the use of medical devices and implants is foreign body associated infection. Its significance in modern medicine and its pathogenesis are discussed. Adherence to and colonization of a foreign body material and the interference of microbial products with the host defence are critical steps leading to foreign body infections. Therapy is difficult and hence the immediate administration of antibiotics and the removal of an infected device or implant is still preferred. New strategies for the prevention of foreign body infections are briefly introduced. PMID- 8407701 TI - Diagnosis and management of the diabetic foot ulcer. AB - Infected leg ulcers in patients with diabetes mellitus are a common and potentially serious problem. Neuropathy and vascular disease associated with diabetes mellitus allow the possibility of significant microbial invasion. Infections in diabetic patients are usually polymicrobial reflecting the normal flora of the foot skin. Curettage of the base of foot ulcers and deep tissue cultures are the most reliable methods for identifying the true pathogens, which are aerobic Gram-negative bacilli, anaerobes, and Staphylococcus aureus. Empirical antibiotic therapy should be directed against these pathogens. Once culture and sensitivity results are available, therapy should be targeted specifically for the pathogens present to prevent long-term use of broad-spectrum antibiotics. Preventive care of the foot in patients with diabetes mellitus is extremely important and may reduce complications associated with infections of the foot. PMID- 8407702 TI - The role of human autoantibodies against gamma-interferon. AB - Natural antibodies to gamma-interferon (IFN-gamma) were detected in the serum of viral infected patients and also in the serum of healthy subjects at a lower titre. The increased titres of antibodies to IFN-gamma in sera from viral infected patients, and their decrease with clinical resolution, indicates that these antibodies are related to the viral infection and probably reflect IFN gamma production as a result of antigenic stimulation in vivo. These antibodies were affinity-purified and studied for their capability to interfere with the multiple activities of IFN-gamma in vitro. Data obtained show that human anti-IFN gamma antibodies interfere with the immunomodulating activity but not with the antiviral and antiproliferative activity of lymphokine. Autoantibodies to IFN gamma may have a role in the immunoregulatory process serving to limit the intensity and or duration of the immune response. Being able to interfere with the immunomodulating activities of IFN-gamma, these antibodies might open up new therapeutic possibilities for those diseases which carry evidence of activated cell-mediated immunity. PMID- 8407703 TI - Panel analysis of the moderating effects of commitment on job satisfaction, intent to quit, and health following organizational change. AB - The authors examined the moderating effects of organizational commitment on the relationship of stress with job satisfaction, intent to quit, and health during organizational turmoil. Panel data were provided by hospital employees surveyed before and after a major divisional consolidation. Findings indicated that commitment buffered the relationship between stress and job displeasure (a canonically derived variate combining residualized job dissatisfaction, intent to quit, and irritation). Stress increased job displeasure only when commitment was low. PMID- 8407704 TI - Enhancement of children's self-esteem through social support training for youth sport coaches. AB - The authors examined the impact of coaching behaviors on players' self enhancement processes. Eight baseball coaches attended a preseason workshop designed to increase their supportiveness and instructional effectiveness. Behavioral guidelines were presented and modeled. A no-treatment control group had 10 coaches. Boys (N = 152) in both groups were interviewed pre- and postseason. Trained coaches differed from controls in player-perceived behaviors in accordance with the guidelines. They were evaluated more positively by their players, their players had more fun, and their teams exhibited a higher level of attraction among players, despite the fact that their teams did not differ from controls in won-lost records. Consistent with a self-esteem enhancement model, findings showed that boys with low self-esteem who played for the trained coaches exhibited significant increases in general self-esteem; low self-esteem youngsters in the control group did not. PMID- 8407705 TI - Generality and specificity in health behavior: application to warning-label and social influence expectancies. AB - The authors outlined a meta-theoretical and an analytic framework for construing the predictive effects of health-behavior expectancies, or beliefs, in terms of both general and specific processes. This framework was applied empirically to the investigation of the predictive effects of outcome expectancies related to the recently mandated alcohol-warning label as well as to expectancies reflecting social influence processes. Results showed that general and specific predictive effects of expectancies on alcohol-use behaviors operated simultaneously, demonstrating the potential value of the framework. The authors summarized implications for continued theoretical development as well as for applications in prevention of alcohol abuse through warning-label and social influence interventions. PMID- 8407706 TI - Job demands and worker health: three-dimensional reexamination of the relationship between person-environment fit and strain. AB - The most influential study of the person-environment (P-E) fit approach to stress was conducted by J. R. P. French, R. D. Caplan, and R. V. Harrison (1982). Unfortunately, this study operationalized fit using various transformations of difference scores, thereby introducing numerous substantive and methodological problems. In the present study, the authors reanalyze data from French et al., using a procedure described by J. R. Edwards (in press) that avoids problems with difference scores and captures the underlying three-dimensional relationship between E, P, and strain. Results resolve ambiguities in the French et al. findings and identify relationships between E, P, and strain that, although consistent with P-E fit theory, cannot be adequately represented by fit measures such as those used by French et al. Implications for P-E fit research are discussed. PMID- 8407707 TI - Heritability of interests: a twin study. AB - The authors administered inventories of vocational and recreational interests and talents to 924 pairs of twins who had been reared together and to 92 pairs separated in infancy and reared apart. Factor analysis of all 291 items yielded 39 identifiable factors and 11 superfactors. The data indicated that about 50% of interests variance (about two thirds of the stable variance) was associated with genetic variation. The authors show that heritability can be conservatively estimated from the within-pair correlations of adult monozygotic twins reared together. Evidence for nonadditive genetic effects on interests may explain why heritability estimates based on family studies are so much lower. The authors propose a model in which precursor traits of aptitude and personality, in part genetically determined, guide the development of interests through the mechanisms of gene-environment correlation and interaction. PMID- 8407708 TI - Neonatal pig pancreatic islets for transplantation. PMID- 8407709 TI - Effect of hemin on growth and DNA synthesis of HL-60 cells. PMID- 8407710 TI - B16 melanoma cells resistant to a novel sialic acid-specific lectin. PMID- 8407711 TI - Framework for validation and implementation of in vitro toxicity tests. AB - The development and application of in vitro alternatives designed to reduce or replace the use of animals, or to lessen the distress and discomfort of laboratory animals, is a rapidly developing trend in toxicology. However, at present there is no formal administrative process to organize, coordinate, or evaluate validation activities. A framework capable of fostering the validation of new methods is essential for the effective transfer of new technologic developments from the research laboratory into practical use. This committee has identified four essential validation resources: chemical bank(s), cell and tissue banks, a data bank, and reference laboratories. The creation of a Scientific Advisory Board composed of experts in the various aspects and endpoints of toxicity testing, and representing the academic, industrial, and regulatory communities, is recommended. Test validation acceptance is contingent on broad buy-in by disparate groups in the scientific community--academics, industry, and government. This is best achieved by early and frequent communication among parties and agreement on common goals. It is hoped that the creation of a validation infrastructure composed of the elements described in this report will facilitate scientific acceptance and utilization of alternative methodologies and speed implementation of replacement, reduction, and refinement alternatives in toxicity testing. PMID- 8407712 TI - Sterol metabolism and oral epithelial cell growth. AB - Previous studies have demonstrated that as the density of cultured oral epithelial cells increases, there is a concomitant increase in phospholipids and cholesterol ester synthesis and a decrease in that of cholesterol and sterol precursors. Other studies have suggested that the effects of exogenous cholesterol sulfate may be similar to growth responses and influence metabolic steps related to cell density. To further examine this possibility, in the present study lipid synthesis was monitored in hamster cheek pouch epithelial cells in cultures established at different cells densities and in the presence of varying amounts of exogenous cholesterol sulfate. Cell [14C]acetate incorporation into lipids was measured in cultures established at four densities ranging from very subconfluent to very dense (postconfluent) in two media, Dulbecco's modified Eagle's medium (DMEM) with 5% fetal bovine serum and KSFM, a non-serum containing keratinocyte medium. Results indicated that the relative proportion of radiolabel incorporated into different lipid classes changed with cell density. In DMEM, the percentage of radiolabel incorporated into total phospholipids and fatty acids increased significantly with increasing cell density whereas percent incorporation into cholesterol, sterol precursors, and cholesterol esters significantly decreased. In KSFM cultures, proportionate phospholipids labeling was significantly increased in more dense cultures whereas cholesterol and cholesterol esters labeling was significantly decreased. In subconfluent and confluent cultures exposed to 10 or 25 microM cholesterol sulfate, the relative proportions of phospholipid labeling also increased significantly compared to dimethyl sulfoxide (solvent) controls, whereas sterol precursors, fatty acids, and cholesterol esters labeling was significantly decreased. These results indicate that cholesterol sulfate can affect cellular lipid synthesis in a manner similar to that which occurs with increasing cell density, and strengthen the hypothesis that cholesterol sulfate may regulate lipid metabolic pathways related to growth and differentiation. PMID- 8407713 TI - Human bone marrow stromal cells express an osteoblastic phenotype in culture. AB - This study reports the selection and characterization of osteogenic precursors from human bone marrow which were isolated by two "clonings" and successive subculturing. These cell lines express alkaline phosphatase activity. Gel electrophoresis of [3H]-proline labeled cultures showed that the cloned cells produce only type I collagen. They synthetize osteocalcin and osteonectin. They respond to 1,25 dihydroxy vitamin D3 by increasing osteocalcin synthesis and secretion, and to parathyroid hormone by increasing cyclic AMP synthesis. After the third subculture in the absence of beta-glycerophosphate, these cell lines formed lots of clusters which exhibit high alkaline phosphatase activity and positive von Kossa staining. X-ray energy spectrum shows that these cells are surrounded by "budding" structures containing calcium and phosphorus with a ratio Ca:P identical to those of pure hydroxyapatite. This process was associated with 45Ca uptake into the cells. All these data support the selection of osteogenic cells which may be of considerable clinical importance. PMID- 8407714 TI - Influence of cell density and receptor number on the binding and distribution of cell surface epidermal growth factor receptors. AB - Previous studies have shown that cell density influences the expression of receptors for at least four growth factors. The data presented in this report demonstrate that epidermal growth factor receptors are regulated differently on cells expressing over a million receptors as opposed to cells expressing approximately fivefold fewer receptors. Specifically, we show that BT-20, MDA 468, and A-431-R1 cells, which exhibit a large number of epidermal growth factor receptors, preferentially down-regulate the high affinity class of these receptors as cell density increases. In addition, we show that these cells express cell surface epidermal growth factor receptors that are localized predominantly to the periphery of the cells. In contrast, A-549 and BSC-1 cells, which exhibit fewer cell surface epidermal growth factor receptors and which reduce all affinity classes of epidermal growth factor receptors as cell density increases, exhibit a diffuse cell surface distribution of these receptors at both low and high densities. PMID- 8407715 TI - Redifferentiation of proliferated rat hepatocytes cultured in L15 medium supplemented with EGF and DMSO. AB - Primary adult rat hepatocytes were cultured in serum-free L15 medium supplemented with 20 mM NaHCO3 and 10 ng/ml epidermal growth factor in a 5% CO2:95% air incubator. The number of cells increased and reached about 180% of the initial value by Day 4, and after 2% dimethyl sulfoxide (DMSO) was added to the culture medium at Day 4, the cells continued to proliferate until Day 6. The number of cells reached about 210% at Day 6 and they were well maintained until Day 18. The cell number gradually decreased with time in culture, but many cells remained for more than 2 mo. On the other hand, without 2% DMSO, the cells proliferated until Day 5, but thereafter they rapidly decreased. After DMSO addition, albumin and transferrin were secreted into the medium and the production of both proteins continued for more than 2 mo. Immunocytochemically both proteins were strongly stained in the cells treated with 2% DMSO. Although the expression of G6Pase in the cells disappeared at Day 6 without DMSO, the cells treated with 2% DMSO recovered G6Pase activity at Day 16. In addition, induction of peroxisomes by 2 mM sodium clofibric acid was clearly shown in the hepatocytes at Day 14 and Day 25 using enzyme-cytochemistry. Ultrastructurally, DMSO-treated hepatocytes had many mitochondria and large peroxisomes with a crystalline nucleoid, and both gap junctions and desmosomes were well developed between the cells even at Day 40. Thus, the number of cells doubled, some differentiated functions of the primary hepatocytes were well restored by the use of 2% DMSO, and these functions were maintained for more than 2 mo. PMID- 8407716 TI - Pluripotent mesenchymal stem cells reside within avian connective tissue matrices. AB - Recent studies have noted the presence of putative stem cells derived from the connective tissues associated with skeletal muscle, heart, and dermis. Long-term continuous cultures of these cells from each tissue demonstrated five distinct phenotypes of mesodermal origin, i.e. muscle, fat, cartilage, bone, and connective tissue. Clonal analysis was performed to determine whether these morphologies were the result of a mixed population of lineage-committed stem cells or the differentiation of pluripotent stem cells or both. Putative stem cells from four tissues (skeletal muscle, dermis, atria, and ventricle) were isolated and cloned. Combined, 1158 clones were generated from the initial cloning and two subsequent subclonings. Plating efficiency approximated 5.8%. Approximately 70% of the 1158 clones displayed a pure stellate morphology, while the remaining clones contained a mixture of stellate, chondrogenic- or osteogenic like morphologies or both. When cultured in the presence of dexamethasone, cells from all clones differentiated in a time- and concentration-dependent manner into muscle, fat, cartilage, and bone. These results suggest that pluripotent mesenchymal stem cells are present within the connective tissues of skeletal muscle, dermis, and heart and may prove useful for studies concerning the regulation of stem cell differentiation, wound healing, and tissue restoration, replacement and repair. PMID- 8407717 TI - Luminol and diazoluminomelanin as indicators of HL-60 cell differentiation. AB - This paper describes use of a novel substituted melanin which is useful in detection of differentiating leukemia cells and their membranes. Comparisons of luminol-(5-amino-2,3-dihydro-1,4-phthalazinedione) and diazoluminomelanin (DALM) mediated chemiluminescence (CL) were made with various types of differentiated and undifferentiated HL-60 whole cells, cell lysates, and membrane fractions. Luminol had a greater CL response than DALM with HL-60 promyelocytic stem cells and differentiated macrophage-like or neutrophil-like whole cell and cell lysate preparations. However, DALM showed markedly greater CL than luminol for membrane fractions derived from each cell type. The greatest luminol-dependent CL was observed for cell types high in myeloperoxidase (MPO). The greatest DALM-mediated CL was seen with cell types that are high in MPO or strong producers of superoxide (O2-) anions. In some cases, significant differences in CL could also be distinguished on the basis of inducing agent used [i.e. dimethylsulfoxide, all trans retinoic acid or 12-o-tetradecanoylphorbol-13-acetate]. Both luminol- and DALM-dependent CL were strongly inhibited by preincubation of cellular preparations with 3-amino-L-tyrosine (a component of DALM). Taken together, these data suggest that the reaction mechanism of luminol favors interaction with cytoplasmic MPO whereas that of DALM favors membrane interactions. Thus, both reagents may be of use in assays to detect differentiating leukocytes or their cellular components. PMID- 8407718 TI - Growth advantage ("clonal dominance") of metastatically competent tumor cell variants expressed under selective two- or three-dimensional tissue culture conditions. AB - In previous experiments it was shown that injection into syngeneic CBA/J mice of cell mixtures containing an excess of non-metastatic SP1 mouse mammary carcinoma cells with a ras transfected metastatic variant of SP1 called C1, always resulted in the eventual dominance of the C1 subpopulation at the site of inoculation. This occurred despite the growth rates of the two cell populations being identical in vivo when grown separately. The means by which the C1 subpopulation achieved "clonal dominance" is thought to involve its responsiveness to stimulatory paracrine growth factors liberated by the non-metastatic SP1 population. The clonal dominance process, however, could not be recapitulated in conventional monolayer tissue culture conditions in which SP1 and C1 cells were grown together in high concentrations of serum, i.e. under non-limiting culture conditions. We now show that clonal dominance of C1 cells can be observed when the cell mixture is maintained in tissue culture for extended periods, or when the cells are grown under selective, limiting conditions, some of which may mimic growth conditions in vivo more accurately. These conditions were a) growth in low (limiting) serum concentrations; and b) growth as three-dimensional multicellular aggregates, i.e. as "tumor spheroids". Under all of these conditions dominance of the C1 subpopulation always took place, but with an efficiency 6- to 40-fold less than generally observed in vivo. C1 cells were also able to form more stable (compact) spheroids compared to SP1 cells. Entrapment of the latter in mixed C1/SP1 spheroids increased the recovery of the SP1 cells suggesting some kind of "rescue" mechanism in which cells are protected from physical forces by three dimensional structure. The relevance of these in vitro interactions for clonal dominance in primary tumors and metastasis in vivo are discussed. PMID- 8407719 TI - Characterization of a cell line derived from zebrafish (Brachydanio rerio) embryos. AB - During the last decade, zebrafish (Brachydanio rerio) have emerged as a novel and attractive system to study embryogenesis and organogenesis in vertebrates. The main reason is that both extensive genetic studies and detailed embryologic analysis are possible using this small tropical fresh water teleost. However, in vitro analysis using cell culture or molecular genetics are still far less advanced than in other vertebrate systems. Here we report the generation and characterization of a fibroblast like cell line, ZF4, derived from 1-day-old zebrafish embryos. The hyperploid cell line has been stable in multiple passages for more than 2 yr now and is the first zebrafish cell line that can be maintained in conventional medium containing mammalian serum. Using a series of plasmids for expression of a marker gene, we evaluate in ZF4 cells the relative strength of expression from several different viral, fish, and mammalian promoters. Stable integration can be obtained by using G418 selection. We hope that our cell line will be a useful tool for the analysis of gene regulation in zebrafish. PMID- 8407720 TI - Brief myocardial ischemia affects free radical generating and scavenging systems in dogs. AB - This study examined whether brief repeated myocardial ischemia altered free radical generating and scavenging activity in a dog model. In dogs preconditioned with four 5-min left anterior descending coronary artery (LAD) occlusions and reperfusions, we examined transcardiac changes in both the function of neutrophils, cells which are major free radical generators, and in myocardial antioxidant enzyme activity, as an indication of free radical scavenging. Neutrophil function was assessed by determining luminol-enhanced whole blood chemiluminescence (CL) induced by zymosan. Blood was taken simultaneously from the carotid artery and the cardiac vein running along the occluded LAD. Preconditioning with sublethal ischemia significantly reduced whole blood CL in the cardiac vein compared with the carotid artery after the first and fourth 5 min reperfusions, while there was no difference in neutrophil count between these sampling sites. Immediately after brief repeated ischemia and reperfusion, manganese-superoxide dismutase (SOD) activity was significantly enhanced, and glutathione reductase activity was markedly reduced in the ischemic, compared with the non-ischemic, myocardium. There were no differences in the myocardial activities of copper, zinc-SOD, glutathione peroxidase, and glutathione S transferase between the ischemic and non-ischemic regions. Also, no difference was observed between the reduced myocardial glutathione levels in these regions, although the oxidized glutathione level was significantly higher in the ischemic regions of the subepicardial and subendocardial areas. We demonstrated that brief repeated ischemia affects free radical generating and scavenging systems in the ischemic myocardium. PMID- 8407721 TI - Involvement of cyclo-oxygenase-generated vasodilating eicosanoid(s) in addition to nitric oxide in endothelin-1-induced endothelium-dependent vasorelaxation in guinea pig aorta. AB - This study investigates the vasodilatory effects of endothelin-1 (ET-1) in isolated guinea pig aortic rings in vitro. Cumulative dose-response curves to ET 1 were constructed and ET-1 actions on prostaglandin F2 alpha (PGF2 alpha) precontraction were studied in both endothelium-intact and endothelium-denuded preparations, in the presence or absence of a cyclooxygenase inhibitor (indomethacin) and/or nitric oxide inhibitors (NG-nitro-L-arginine methyl ester and hemoglobin). In endothelium-intact preparations, pretreatment with indomethacin (10(-5) M, 30 min), alone or in combination with NG-nitro-L-arginine methyl ester (L-NAME, 10(-4) M), significantly augmented the constrictive responses to ET-1, whereas indomethacin, L-NAME, and hemoglobin (10(-5) M) had no significant effects in the endothelium-denuded preparations. Furthermore, in PGF2 alpha-precontracted, endothelium-intact preparations, ET-1, at a dose of 10(-9) M, induced initial relaxation followed by subsequent contraction, while it only contracted the endothelium-denuded preparations. The initial relaxation was abolished by indomethacin, but not by L-NAME or hemoglobin. In addition, this relaxation was not inhibited by a specific ETA receptor antagonist, BQ-123 (6 x 10(-6) M). In addition to the involvement of nitric oxide, these results show the involvement of cyclo-oxygenase-generated vasodilating eicosanoid(s) derived from endothelium in ET-1-induced vasorelaxation in guinea pig aorta in vitro. The results also indicate that this vasorelaxation is mediated by ETB receptor activation. PMID- 8407722 TI - Effects of pentobarbital on inotropic state of isolated canine left ventricle. AB - Although pentobarbital has been found to depress myocardial function, the magnitude of its direct effects on ventricular contraction at anesthetic concentrations has not been well quantified. The direct effects of pentobarbital on left ventricular function were measured by employing an isolated canine heart preparation with a blood oxygenator. Seven hearts were perfused with blood, dextran, and perfluorochemical artificial blood. Ventricular function was evaluated using the slope of the end-systolic pressure-volume relationship (Ees) and the maximal rate of pressure development (dP/dtmax) in ventricles contracting isovolumically in control, after a low dose (13 micrograms/ml), and after a high dose (48 micrograms/ml) of pentobarbital. These concentrations represent one-half and two times the typical value (25 micrograms/ml) found to produce anesthesia in canines (assessed by tail clamp or blink reflex). The low dose of pentobarbital did not produce clear-cut depression in contractile function. The high dose of pentobarbital produced significant reductions of Ees, and dP/dtmax: Ees decreased 29%, from a control of 4.30 +/- 0.84 to 3.05 +/- 0.49 mmHg/ml and dP/dtmax decreased 24%, from a control of 909 +/- 148 to 695 +/- 173 mmHg/s. Thus, the threshold for the direct depressant effect of pentobarbital on ventricular function falls within the range of half to double the typically-reported anesthetic concentrations. PMID- 8407723 TI - Characteristics of single isovolumic left-ventricular pressure waves of dog hearts in situ. AB - By fitting isovolumic phases of an ejecting beat with a model-wave function, one can predict source pressure of the ejecting beat (Sunagawa et al. Trans Biomed Eng 1980; 27:299-305), this being a major determinant of systolic performance. Prior applications of this principle have involved two assumptions: (1) that the isovolumic pressure wave is shaped like an inverted cosine wave, and (2) that duration of an isovolumic beat is the same as that of an ejecting beat. The first assumption might cause overestimation of source pressure, since an isovolumic pressure wave begins declining before the midpoint of the wave. The second assumption might cause underestimation of source pressure, since an ejecting beat is always shorter than an adjacent isovolumic beat at the ejecting beat's end diastolic volume. Although the two errors tend to cancel, it would be more rational and accurate to use a realistic model wave shape and a realistic isovolumic beat duration. To acquire the information necessary for this, pressure and volume time courses were measured during ejecting beats and adjacent isovolumic beats in dogs under the following steady-state conditions: basal, atrial pacing at various rates, infusion of dobutamine, infusion of verapamil, coronary ligation(s), and ventricular pacing at various sites. These conditions affected the amplitude and duration of isovolumic pressure waves substantially but did not affect the shape of the waves significantly. The duration of each isovolumic beat exceeded that of the previous ejecting beat to a degree which corresponded approximately to the ejecting beat's normalized pressure reserve (source pressure minus peak ejection pressure)/(source pressure). A more accurate source-pressure prediction should be possible by use of a realistic isovolumic pressure-wave shape and by taking account of the effect of pressure reserve on contraction duration. PMID- 8407724 TI - Morning increase in hemodynamic response to exercise in patients with angina pectoris. AB - The present study was conducted to determine whether or not there is diurnal variation in the hemodynamic responses to stimuli that increase myocardial oxygen demand, and the effects of such variation on electrocardiograms (ECG). Fifteen patients with angina pectoris, 17 patients with old myocardial infarction, and 8 healthy controls were examined in this study. Graded exercise stress testing was conducted in the supine position, once in the morning and once in the afternoon, using a bicycle ergometer. A standard 12-lead ECG was recorded before, immediately after, and 3, 5, and 10 min after the end of the exercise. The exercise ECG and blood pressure changes were compared among the groups and, within each group, the results after morning and afternoon exercise were compared. Hemodynamic responses, including heart rate, blood pressure, and the pressure-rate product, showed greater increases in the morning than in the afternoon in angina patients and controls, in association with greater depression of the electrocardiographic ST-segment. In contrast, patients with old myocardial infarction exhibited no difference in hemodynamic responses or the ST-pattern from morning to afternoon. The results suggest that diurnal variation of hemodynamic responses to increased oxygen demand may explain, at least partly, why myocardial ischemia of effort angina is more severe in the morning than in the afternoon. PMID- 8407725 TI - Cardiomyopathy of glycogen storage disease type III. AB - To identify the severity of cardiac involvement in glycogen storage disease type III (GSDIII), and its relation to skeletal muscle involvement and age, 23 patients were studied. The median age was 10 years. Echocardiography, electrocardiography, and creatine phosphokinase (CK) levels were used to assess cardiac and skeletal muscle involvement. Septal and left ventricular posterior wall measurements were compared with normal data. Shortening fraction was derived from left ventricular cavity dimensions. In some patients the echocardiogram resembled that of hypertrophic cardiomyopathy. Thirteen of 20 electrocardiograms (ECG) were abnormal. Eleven patients had septal and/or posterior wall thickness > 95% confidence limits (CL). Despite this, cardiac symptoms were uncommon. The CK levels were not directly associated with cardiac abnormalities. Older patients (> 20 years) had more abnormal measurements of posterior wall thickness than did younger ones (< 20 years). This finding, albeit in a cross-sectional series, suggests progressive myocardial involvement with age despite the absence of symptoms. PMID- 8407726 TI - Immunohistochemical localization of plasminogen activator inhibitor-1 in human coronary atherosclerotic lesions involved in acute myocardial infarction. AB - Immunohistochemical localization of plasminogen activator inhibitor-1 (PAI-1) was studied using the streptavidin-biotin method in human atherosclerotic coronary arteries. The patients (one male and two females), whose ages ranged from 61 to 78 years, died of anteroseptal acute myocardial infarction without having received any thrombolytic therapy. PAI-1 immunoreactivities (IRs) were mainly detected in the endothelial cells, smooth muscle cells, and collagen fibers of the coronary arterial intima and not in thrombi. Remarkable immunohistochemical staining was seen in intimal collagen fibers. In one patient, intimal PAI-1 IRs partly bordered a thrombus and surrounded a large atheroma rich in cholesterol crystals. Our results suggest that PAI-1 is present in both cellular and extracellular components of human coronary atherosclerotic lesions. PMID- 8407727 TI - An autopsy case of intracranial hemorrhage during tissue plasminogen activator infusion. AB - We describe an autopsy case of severe intracranial hemorrhage which occurred during the infusion of tissue plasminogen activator (t-PA) for acute myocardial infarction. A 75-year-old man was admitted with substernal chest pain of 3-h duration and electrocardiographic changes consistent with an acute inferior myocardial infarction. Physical examination was unremarkable, except for an initial blood pressure reading of 160/96 mmHg. The patient received 3,000 IU intravenous heparin followed by a 2.4 x 10(6) IU bolus dose of tissue plasminogen activator (t-PA) (Alteplase). This was followed by a drip infusion of 21.6 x 10(6) IU of t-PA over 1 h (total dose 41 mg). Thirty minutes after the infusion of t-PA was initiated, the patient suddenly lost consciousness and began to have violent convulsions, followed by cardiac arrest. Autopsy revealed massive hemorrhage in the bilateral cerebrum and brain stem. To our knowledge, this is the first case of sudden death during t-PA infusion therapy. PMID- 8407729 TI - New approaches. PMID- 8407728 TI - Rapidly growing mural thrombus in an abdominal aortic aneurysm. AB - While mural thrombus accompanied by an abdominal aortic aneurysm (AAA) is not rare, the growth rate of such a thrombus has not yet been adequately documented. We present here a very rare case of a 62-year-old female patient with an AAA in whom the mural thrombus in the aneurysm grew very rapidly over a short period. We could follow the growth of the mural thrombus in the AAA by two-dimensional (2-D) abdominal echography. Patients with an AAA must be closely monitored by this technique which is able to detect the presence of the thrombus and allow evaluation of its growth. PMID- 8407730 TI - Latex: a new occupational hazard for physicians. AB - Identification of latex sensitive individuals can be life saving. The American College of Allergy and Immunology has put forth the following recommendations: 1) Just as patients are routinely asked preoperatively about allergy to medications prior to treatment, a careful history should be obtained from every patient prior to any procedure involving contact with latex. Any patient who has a history of rash, itching, hives, rhinitis, swelling, or eye irritation or asthmatic symptoms after touching a balloon, rubber glove or any latex containing object is at risk for anaphylaxis. Previous medical history, of unexplained allergic or anaphylactic reactions during a medical procedure, may indicate sensitization. 2) Health care providers or other workers who give a history of only mild latex glove eczema rarely have anaphylactic events. However, a history of work-related conjunctivitis, rhinitis, asthma, or urticaria may indicate allergic sensitization and increased risk for more severe reactions in the future. 3) The utility of screening tests for predicting anaphylaxis remains controversial. Suitable reagents for skin prick tests for latex are not commercially available at this time. Inasmuch as prick testing appears to carry a substantial risk of inducing anaphylaxis, this test must be considered experimental and it should only be done by experienced physicians with resuscitative equipment and personnel immediately available. Serum tests for latex-specific IgE, currently performed on a research basis at several laboratories, may confirm a suspected diagnosis in many cases but presently lack sensitivity to identify all patients with true latex allergy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407731 TI - Loose body of the elbow mimicking tumor. AB - Distinguishing between calcific soft-tissue masses and intra-articular loose bodies about the elbow can be difficult. We present the case of a 59-year-old woman with a slowly enlarging soft-tissue mass about the elbow. History, physical examination, and diagnostic studies did not establish a definite diagnosis, nor did they rule out the possibility of a neoplastic process. Excisional biopsy revealed a loose body contained in a cystic mass with elbow joint communication. PMID- 8407732 TI - Radiological case of the month. Normal epicardial fat pad. PMID- 8407733 TI - Asthma and allergy: some answers and more questions. PMID- 8407734 TI - The interrelationship among allergy, airways responsiveness, and asthma. PMID- 8407735 TI - Educating school personnel about asthma. AB - Teachers, administrators, and other nonmedical personnel are frequently responsible for managing students with asthma during school hours. We determined that the school secretary is the most likely person to manage asthma at school. We developed an educational program for both nonmedical and medical school personnel entitled "Asthma Management in the Schools." A questionnaire designed to evaluate knowledge and obtain information about asthma care at school was mailed to participants several weeks before they attended the program and was administered again after the program was presented. Nonmedical personnel had lower mean preclass test scores than nurses, but mean postclass test scores were similar. Programs designed to improve asthma care in school should meet the needs of nonmedical personnel. PMID- 8407736 TI - A comparison of patient attitudes toward asthma self-management among acute and preventive care settings. AB - This study characterizes the attitudes regarding asthma and asthma care of low income, African-American adults who receive care from acute care settings. As a point of reference, their attitudes and knowledge were compared with those of a group of patients receiving asthma care from a private setting that stresses preventive asthma self-management. Patients were assessed regarding attitudes toward (1) routine asthma self-care and decisions as to when to self-treat versus seek asthma care, (2) administration of asthma medications, (3) satisfaction with acute-care services, and (4) the desire for asthma education. Asthma morbidity, sociodemographic characteristics, awareness of environmental triggers, and daily stressors were also assessed. Responses of adults receiving most of their asthma care from acute care settings suggest the same pattern of failure to treat asthma regularly and delay in seeking care as implicated in asthma deaths among children. Emphasis on self-treatment of asthma symptoms, not preventive self management, was apparent among the attitudes of the acute care patients. Lack of regular care, delay of treatment, and reliance on self-treatment via over-the counter medications was noted. Implications of these findings for the development of asthma education programs are addressed. PMID- 8407737 TI - Evaluation of the Autohaler actuator: the effect of written patient instructions on correct use. AB - The Autohaler actuator is a breath-actuated device designed by 3M Pharmaceuticals. The objective of this study was to measure and improve the effectiveness of the device's package insert instructions (PII). Using only the PII for guidance, 5 of 20 (25%) subjects failed to trigger the device. The PII were revised based on the subject's performance and suggestions. Using the revised PII, only 1 of 20 (5%) different subjects failed to trigger the device. These results indicate that relatively minor modifications to instructions can result in significant improvements in patient use. Eighty-five percent of the participants thought the device was easier to use than a metered-dose inhaler. PMID- 8407738 TI - The effect of nedocromil sodium on histamine responsiveness in clinically stable asthmatic children. AB - A double-blind, randomized, placebo-controlled, parallel-group study was undertaken in 120 clinically stable asthmatic children (aged 6-19 years) to determine the ability of 8 weeks of treatment with nedocromil sodium (4 mg, three times a day) to reduce the level of histamine responsiveness. Despite the subjects being clinically stable and reporting few asthma symptoms, approximately one-third had abnormal pulmonary function on enrollment into the study. Statistically significant increases in pulmonary function were seen in the group treated with nedocromil sodium but not in the control group (p = 0.01). Furthermore, 52% of the individuals with abnormal pulmonary function returned to normal following treatment with nedocromil sodium compared to 11% of those with abnormal pulmonary function who received placebo. However, there were no differences in the level of histamine responsiveness between the two treatment groups at baseline or after 4 or 8 weeks of treatment with nedocromil sodium or placebo. These data suggest that the level of histamine responsiveness is not intimately related to the level of asthma control in children. PMID- 8407739 TI - Reduction of secondary smoke exposure in asthmatic children: parent counseling. AB - Epidemiological evidence shows that children's exposure to secondhand tobacco smoke increases their risk of respiratory illness. This study evaluated five families and their asthmatic children (aged 5-14 years) in an outpatient counseling program for reducing the children's exposure to passive smoking. Intervention included biweekly counseling/instructions for parents to limit their children's tobacco exposure. A multiple-baseline, quasiexperimental design was used for self-reported measures of the children's smoke exposure and the parent's smoking frequency. Counseling was associated with smoke exposure reduction of 40 80% from baseline for each of 5 children, with most improvements sustained during follow-up. This study provides support for the development of tobacco exposure prevention programs for children with pulmonary disease. PMID- 8407740 TI - An increase in superoxide generation of bronchoalveolar lavage fluids in the model of late asthmatic response in guinea pigs. AB - To clarify the possible role of superoxide anion of bronchoalveolar cells in the pathogenesis of late asthmatic response (LAR), I performed an allergen inhalation test using a guinea pig LAR model and estimated subsequent changes in activities of superoxide generation in bronchoalveolar lavage fluids (BALFs). Significant increases in superoxide generation of BALFs occurred not only immediately, but 6 hr after ovalbumin inhalation challenge in the LAR model, and the increased levels were prolonged 24 hr later. The present results suggest that significant increases in superoxide anion of the respiratory cells were involved in the antigen-induced late responsiveness of bronchial asthma. PMID- 8407741 TI - Vocal cord dysfunction in an asthmatic child: case report. AB - Acute functional narrowing of the glottis frequently leads to an inappropriate diagnosis of asthma. Only 2 cases have been reported of patients with asthma and concomitant vocal cord dysfunction. We present the case of an adolescent boy with asthma, who exhibited a worsening of wheezing and a reduction in peak expiratory flow rate out of proportion to symptoms due to a vocal cord dysfunction. PMID- 8407742 TI - Severe acute asthma followed by quick reversal. PMID- 8407743 TI - A psychiatric population of prenatally cocaine-exposed children. AB - The backgrounds and behaviour of 40 prenatally cocaine exposed children referred to the child psychiatry program of an inner-city teaching hospital are explored. The population described is unique to the literature in these ways: (1) Most of the children were not detected as cocaine-exposed at birth, and were disclosed as cocaine-exposed by historical account. (2) Most of the children had lived within drug-dominated home environments for varying lengths of time. Findings indicated that the children receiving intervention early in life, whether detected as cocaine-exposed at birth or by historical account, fared much better socially and emotionally as measured by rate of abuse and severity of psychiatric symptoms. It is concluded that instances of prenatal cocaine exposure should be ascertained at the earliest possible point to intervene in potentially deleterious environmental outcome and associated psychiatric morbidity. Four illustrative cases are presented. PMID- 8407744 TI - Follow-up of psychiatric and educational morbidity among adopted children. AB - OBJECTIVE: The purpose of this study is to evaluate longitudinally the strength of association between adoptive status and psychiatric and educational morbidity and substance use. METHOD: This study makes use of data from the 1983 Ontario Child Health Study and 1987 follow-up. This community survey of children (4- to 16-years-old in 1983, 8- to 20-years-old in 1987) included a subpopulation of adopted children. The primary outcomes measured were psychiatric disorder, poor school performance, and substance use. RESULTS: Adoption, identified in 1983, in boys was a significant marker for psychiatric disorder in 1987. Adoption was not a significant risk indicator for educational morbidity or substance use in 1987. In the multivariate analyses, adoptive status demonstrated no independent influence on 1987 educational morbidity or substance use. However, adoptive status, in the presence of poor school performance in 1983, was a significant risk indicator for psychiatric disorder in 1987. CONCLUSIONS: Adopted children did not do significantly worse than nonadopted children over time in terms of educational morbidity or substance use, but adopted boys demonstrated a significantly increased risk of psychiatric disorder versus nonadopted boys. PMID- 8407745 TI - Long-term effects of maternal loss on Vietnamese Amerasians. AB - OBJECTIVE: The objective of this study was to investigate the differential effects of maternal, single surrogate, and multiple surrogate caregiving on psychological and educational outcomes in a group of Vietnamese Amerasians. METHOD: Subjects were assessed in Vietnam using several measures of affective and behavioral symptomatology as well as a detailed psychosocial history. RESULTS: Amerasians who lived continuously with their biological mothers had fewer symptoms of psychological distress than did either group raised by surrogate caregivers. In addition, those living continuously with either their mothers or a single surrogate had better educational outcomes and experienced fewer geographic relocations than did those raised by multiple surrogates. CONCLUSIONS: Amerasians who remained continuously with their biological mothers had better overall outcomes than did those raised by surrogate caregivers. Continuous surrogate caregiving appeared to offer some, but not complete, protection from the adverse effects of maternal loss. The clinical significance of these findings for those assessing and treating Amerasians, as well as for those assisting immigration officials with relocation decisions, is discussed. PMID- 8407746 TI - Role of parent-child relationships in mediating the effects of marital disruption. AB - OBJECTIVE: This study sought to explore the long-term effects of and relationships between parental divorce, interparental conflict, and parent-child relationships on interpersonal and generalized trust, empathy, dependency, and depressive affect among late adolescents. METHOD: Self-administered questionnaire data were collected from a sample of 288 college students (including 60 children of divorce). Participants were asked to rate current and past levels of interparental conflict, the current affective quality of their parent-child relationships, and present adjustment. RESULTS: Path analyses revealed that the effects of interparental conflict on psychological well-being were mediated by parent-child relationships. For women, however, parental divorce also was found to affect adjustment indirectly (via disrupted father-daughter relationships), although no such path emerged for men. CONCLUSIONS: The findings are part of a growing literature documenting the role of parent-child relationships in mediating the effects of marital disruption. In addition to efforts to resolve marital conflict, clinicians may want to focus on enhancing mother-child, father child relationships as a way to modify the damaging effects of conflictual marital relations on children and adolescents. PMID- 8407747 TI - The written word in psychotherapy with a latency age girl. AB - Play therapy is the treatment of choice with young children, while "talking" therapy is used with older children and adults. There have been reports of written communication being employed in therapy both with children (e.g., diaries and stories) and with adults (e.g., letters and contracts), but little has been reported about the use of the exchange of written words during therapy sessions with "literate" children. The author presents segments of the treatment of a latency age girl to illustrate how the exchange of written language can allow treatment to proceed when children are too distressed to play or to talk at certain points in therapy. Samples of this communication are offered. PMID- 8407748 TI - Are perinatal complications relevant to the manifestation of ADD? Issues of comorbidity and familiality. AB - OBJECTIVE: We evaluated the role of pregnancy, delivery, and infancy complications (PDICs) in the etiology of attention deficit disorder (ADD) addressing issues of comorbidity and familiarity by formulating and testing multiple hypotheses. METHOD: Subjects were six to 17-year-old boys with DSM-III attention deficit disorder (ADD, N = 73), psychiatric (N = 26), and normal (N = 26) controls and their relatives. Information on PDICs was obtained from the mothers in a standardized manner blind to the proband's clinical status. RESULTS: Using odds ratio analyses, an association was found between ADD and PDICs that was strongest for the comorbid and nonfamilial subtypes. In contrast, noncomorbid and familial ADD subgroups differed less from normal controls in the risk for PDICs. CONCLUSIONS: The increased risk for PDICs in nonfamilial ADD children and the lack of evidence for increased risk among familial ADD patients suggests that PDICs may be part of nongenetic etiologic mechanisms in this disorder, especially for children who have comorbid disorders. PMID- 8407749 TI - Discriminant validity of attention-deficit hyperactivity disorder. AB - OBJECTIVE: The primary purpose of this study was to assess the discriminant validity of attention-deficit hyperactivity disorder (ADHD) relative to well defined groups of children with other psychiatric disorders. METHOD: Clinic referred patients diagnosed as having ADHD without any other comorbid diagnosis (N = 13), were compared with patients with only anxiety disorders (N = 20), disruptive disorders other than ADHD (N = 15), and nonreferred controls (N = 18) on measures of cognitive and academic functioning, as well as on objective measures of attention, impulse control, and activity level. RESULTS: All three patient groups were found to have cognitive and academic achievement difficulties relative to controls. However, the ADHD group was found to be inattentive and impulsive relative to the other patient groups and the nonreferred controls. Objective measurement of activity level distinguished the ADHD group from controls but not from the other two patient groups. CONCLUSIONS: These data support the diagnostic validity of a small subgroup of ADHD children (i.e., those without comorbid diagnoses) and demonstrates, that as a group, these children can be distinguished from patients with anxiety as well as other disruptive disorders on objective test measures. PMID- 8407750 TI - Familial relationship between Gilles de la Tourette's syndrome, attention deficit disorder, learning disabilities, speech disorders, and stuttering. AB - OBJECTIVE: This study examined the familial relationship between Gilles de la Tourette's syndrome (GTS) and attention deficit disorder (ADD), learning problems, speech disorders, and stuttering. METHOD: This family study consisted of 338 first degree relatives of 85 GTS probands and 113 controls. All available relatives were personally assessed using structured interviews, and family history information was collected from all family members. Best-estimate diagnoses were assigned for GTS, ADD, learning problems, speech disorders, and stuttering. Analyses examined whether ADD, learning problems, speech disorders, and stuttering by themselves represent genetically variant expressions of GTS. RESULTS: There was no evidence that ADD, learning problems, speech disorders, or stuttering represent variant expressions of GTS. However, relatives with GTS were at increased risk for ADD regardless of the ADD diagnosis of the GTS proband. CONCLUSIONS: ADD, learning problems, stuttering, and speech problems by themselves are not variant forms of GTS. However, GTS and ADD may be etiologically related in some persons. There may be two types of individuals with GTS and ADD: ones in whom ADD is independent of GTS, and others in whom ADD is secondary to occurrence of GTS. PMID- 8407751 TI - Symptoms of attention-deficit hyperactivity disorder in an Italian school sample: findings of a pilot study. AB - OBJECTIVE: The objective of this study was to complete a teacher questionnaire on a sample of children (N = 232) in nine fourth grade classes in schools in two regions of central Italy to assess the frequency of occurrence of symptoms of attention-deficit hyperactivity disorder (ADHD) and the rates of probable cases in the sample. METHOD: Each ADHD symptom was rated by the teacher as either absent (0), sometimes present (1) or frequently present (2). RESULTS: Of the children 3.9% had eight or more DSM-III-R Criterion A symptoms of ADHD scored at a "frequent" level (score of 2) and were considered to be "likely cases" of ADHD; an additional 6.9% did not meet this threshold but had a total score of 16 or more on the scale and were considered to be "possible cases." CONCLUSIONS: The findings suggest the need for more systematic epidemiological investigations to evaluate the true prevalence of the syndrome and its risk factors in the Italian population. PMID- 8407752 TI - Social Adjustment Inventory for Children and Adolescents: concurrent validity in ADHD children. AB - OBJECTIVE: The assessment of children's adaptive functioning has broad-range implications for clinical practice, research, and public health, and relatively few well-developed assessment instruments are available. The validity of the Social Adjustment Inventory for Children and Adolescents (SAICA), a semistructured interview schedule that assesses adaptive functioning in children and adolescents, was examined. METHOD: Two groups of index children were examined: 140 attention-deficit hyperactivity disorder (ADHD) children (both psychiatric and pediatric referrals) and 120 normal comparisons. RESULTS: Concurrent validity was found for the SAICA in its significant association with the social competence scales of the Child Behavior Checklist and DSM-III-R's Global Assessment of Functioning. It was also found that among ADHD children, psychiatric comorbidity conferred an increased risk for worse adaptive functioning. CONCLUSIONS: These findings suggest the SAICA is an effective way to assess adaptive functioning. Moreover, its ability to discriminate ADHD from control children and its association with psychiatric comorbidity indicate the SAICA measures adaptive functioning within a range relevant for studies of psychopathology. PMID- 8407753 TI - Side effects of methylphenidate and desipramine alone and in combination in children. AB - OBJECTIVE: To investigate side effects of methylphenidate and desipramine alone and in combination in hospitalized children with symptoms of attention-deficit hyperactivity disorder and depression. METHOD: A double-blind placebo controlled crossover design was used to investigate each medication alone and in combination. Side effect ratings and EKGs were done weekly. Pulse and blood pressure were monitored daily. RESULTS: Nausea, dry mouth, and tremor were present in at least twice as many children on combined methylphenidate and desipramine compared with any other condition. Nausea/vomiting, headaches, other aches, refusal of food, and feeling "tired" were significantly more frequent during the combined methylphenidate plus desipramine condition when compared with either methylphenidate alone or with baseline. Significantly higher ventricular heart rate was found on combined methylphenidate plus desipramine compared with desipramine alone, methylphenidate alone, and baseline. Prolonged PR interval and significantly higher heart rate occurred during desipramine alone compared with baseline. CONCLUSIONS: During the several-month duration of the study, there were more frequent side effects during combined methylphenidate plus desipramine treatment than with either medication alone. Clinically, side effects present during combined medication appeared to be similar to and no more serious than those associated with desipramine alone. PMID- 8407754 TI - The gene for ADHD? Not yet! PMID- 8407755 TI - EPS with lithium. PMID- 8407756 TI - Fluoxetine and memory impairment. PMID- 8407757 TI - Negative results with clomipramine. PMID- 8407758 TI - Brain SPECT imaging. PMID- 8407759 TI - Important dose correction. PMID- 8407760 TI - Is elective mutism a social phobia? PMID- 8407761 TI - Child sexual abuse: immediate and long-term effects and intervention. AB - OBJECTIVE: The primary purpose of this review is to highlight the progress made in the area of child sexual abuse during the recent decade and to identify the gaps in our current knowledge about this syndrome. METHOD: More than 100 articles on child sexual abuse were reviewed, the majority written from 1980 to the present concerning the demographics of child sexual abuse, the psychological effects of child sexual victimization, the psychopathology encountered in adult survivors of child sexual abuse, hypotheses regarding the nature of the trauma, a critique of the research, and approaches to intervention. RESULTS: Although a wide variety of psychological sequelae have been documented in sexually abused children referred for evaluation or treatment, there appears to be considerable variability in the severity of the symptoms, and we remain ignorant of sequelae in abused children who never enter the mental health system. However, some of these children may become symptomatic in adult life. Validation of sexual abuse is hampered by the lack of specific behavioral markers. Methodological difficulties in child sexual abuse research include problems with definition, failure to measure severity of the abuse, sampling problems, failure to use standardized or appropriate instruments, problems with validation, and failure to use control groups. CONCLUSIONS: Despite an increased focus on child sexual abuse in the recent decade, many gaps remain in our knowledge. Prospective longitudinal follow-up studies of sexually abused children and treatment outcome studies are urgently needed. PMID- 8407762 TI - False statements and the differential diagnosis of abuse allegations. AB - OBJECTIVE: Because child psychiatrists do not have a consistent way to classify the untruthful child and because there are no generally accepted definitions of the many ways in which false statements occur in allegations of abuse, the objective of this paper is to classify and define the various ways in which false statements occur in allegations of abuse. METHOD: The author reviewed 40 articles, chapters, and books that contained examples of false statements made by children or caregivers in the context of an abuse allegation. RESULTS: This paper clarifies the concepts of indoctrination, suggestion, fantasy, delusion, misinterpretation, miscommunication, innocent lying, deliberate lying, confabulation, pseudologia phantastica, overstimulation, group contagion, and perpetrator substitution. CONCLUSION: The correct classification of abuse allegations is important in both clinical and forensic child psychiatry. The definitions in this paper, which are based on clinical experience, should be studied through systematic research. PMID- 8407763 TI - Prevalence of childhood sexual abuse experiences in a community sample of women. AB - OBJECTIVE: The study was designed to ascertain the prevalence and nature of sexual abuse in childhood for a community sample of women. METHOD: A two-stage design, using questionnaires and face-to-face interviews, was employed, providing information on prevalence rates, types of abuse, ages of victims, relationship to the abuser, and cohort effects. RESULTS: Nearly one woman in three reported having one or more unwanted sexual experiences before age 16 years. A significant number of these experiences (70%) involved genital contact or more severe abuse, and 12% of those abused were subjected to sexual intercourse. The abusers were usually known to the victim, being family members in 38.3% of cases and acquaintances in another 46.3%. Stranger abuse accounted for 15% of all abuse experiences. Most of the abusers were young men, disclosure of the abuse was infrequent, and only 7% of all abuse was ever officially reported. Prevalence rates showed no urban/rural differences, no cohort effect with subject age, and no age differences in disclosure rates. CONCLUSIONS: Child sexual abuse is common, serious, infrequently reported, and the abuser is usually known to the child. Preadolescent girls are at greatest risk. PMID- 8407764 TI - Developmental differences in detection and disclosure of sexual abuse. AB - OBJECTIVE: This study examined the role of developmental differences in the detection and disclosure of child sexual abuse. METHOD: A random medical record review was conducted of 72 children and adolescents seen over a consecutive 3 year period for suspected sexual abuse in the emergency room of a pediatric hospital. The following data were gathered: (1) demographic information, (2) presenting symptomatology that initiated caregiver decision to seek evaluation, (3) type of disclosure (purposeful or accidental), and (4) precipitants to disclosure. RESULTS: Preschool age children were significantly more likely than school age children and adolescents to exhibit behavioral or physical symptoms that prompted caregivers' suspicion of sexual abuse. Preschool age children made disclosures accidentally and typically with an immediate precipitating event unrelated to the abuse itself. In contrast, sexual abuse disclosures from school age children were purposeful and not associated with a precipitating event. CONCLUSIONS: Practitioners need to be aware of the developmental differences in the detection of sexual abuse, including the forces that inhibit disclosure in older children. PMID- 8407765 TI - Clinical expertise and the assessment of child sexual abuse. AB - OBJECTIVE: Mental health specialists (N = 48) were surveyed as to (1) their estimates of the likelihood that a 3-year-old child had been sexually molested (as alleged by her mother in the context of a child custody dispute) by her father, and (2) their recommendations, given their estimates, as to child visitation/custody. METHOD: Specialists heard a detailed presentation of the court-appointed clinician's findings in this case, which included parent interviews and videotaped child-parent interaction sequences. RESULTS: The array of estimated likelihoods was extreme despite that all the clinicians heard the same case. Recommendations to the court strongly tended toward restriction of child-father contact, even when estimates of the likelihood of abuse were low. CONCLUSIONS: Courts should be highly cautious in relying on clinical experts in child custody cases entailing allegations of child sexual abuse. Practitioners should be candid with courts concerning the absence of diagnostic precision in such cases. PMID- 8407766 TI - Evaluation of risk factors in childhood sexual abuse. AB - OBJECTIVE: This study evaluated whether the level of distress in sexually abused girls was predicted by the relationship of the perpetrator to the victim, the kind of abuse, the use of force, removal from the home, and race or ethnicity. METHOD: Seventy-five girls, recently reported as having been sexually abused, completed measures on depression, anxiety, and self-worth, which were then trichotomized into distress levels. RESULTS: Penetration predicted higher levels of distress on depression and self-worth measures. Force predicted higher levels of distress on those measures when the perpetrator was not a father figure and lower levels of distress when the abuser was a father figure. CONCLUSIONS: These differential effects may be related to issues of self-blame and responsibility that vary with the relationship of the perpetrator and may be useful in developing interventions. PMID- 8407767 TI - Cross-gender behavior and gender conflict in sexually abused girls. AB - OBJECTIVE: This study contrasted a group of sexually abused girls, aged 6 to 12 years, with two demographically comparable control groups, girls from a child psychiatry outpatient clinic, and girls from a general pediatric clinic, to determine whether differences in gender role behavior and identity could be demonstrated. METHOD: All girls underwent an evaluation protocol that included a semistructured interview for children, the Gender Role Assessment Schedule--Child (GRAS-C). The mothers were administered several questionnaires including two parent-report measures of gender-related behavior in their children, the Child Game Participation Questionnaire (CGPQ) and the Child Behavior and Attitude Questionnaire--Female version (CBAQ-F). RESULTS: Sexually abused girls manifested significantly more cross-gender behavior on the GRAS-C (in the areas of gender role preference and aggression) and gender identity conflict than did nonabused girls in both comparison groups. For the sexually abused and psychiatric control groups, their parents reported greater involvement in traditionally masculine games on the CGPQ, but on the CBAQ-F, no significant group differences were found. CONCLUSIONS: Findings suggest that sexual abuse in preadolescent girls is associated with cross-gender behavior and gender conflict. PMID- 8407768 TI - Predictors of self-reported psychopathology in children abused repeatedly by a parent. AB - OBJECTIVE: This study was designed to test the influence of gender, type of abuse, and other factors on the development of psychopathology in abused children. METHOD: Forty-one children who had been abused repeatedly either physically or sexually by a parent or parent figure were given the revised Diagnostic Interview for Children and Adolescents. RESULTS: Logistic regression analyses showed: conduct disorder was predicted by male gender and increasing age regardless of type of abuse; post-traumatic stress disorder was predicted by number of stressors other than abuse and not by sexual abuse versus physical abuse. Predictors of somatization, psychotic symptoms, suicidal ideation, and separation anxiety also were identified. CONCLUSIONS: Gender, age, and stressors other than abuse contribute to the prediction of psychiatric disorder in abused children. Early interventions with young abused boys could help prevent development of violent behavior or conduct disorder; case work to reduce other stressors and treatments designed to reduce the impact of stressors may ameliorate developing post-traumatic stress disorders and other emotional disorders in abused children. PMID- 8407769 TI - Psychological distress, problem behaviors, and family functioning of sexually abused adolescent inpatients. AB - OBJECTIVE: By studying children and adolescents who have been sexually abused, improvements in identifying and treating emerging pathology can be gleaned before personality structures become more rigidly defined and less amenable to therapeutic interventions. It is through studying and comparing subgroups of victims and controls that clinical researchers can additionally elucidate abuse related manifestations that contribute to the development of psychopathology. METHOD: A group of sexually abused adolescent psychiatric inpatients was compared with a control group of inpatient counterparts on measures of social competence, self-esteem, depression, substance abuse, and perceptions of family characteristics and functioning. Consecutive admissions to an adolescent inpatient psychiatric unit (N = 423) were carefully screened for a history of sexual abuse, then divided into two groups, each with 87 subjects, after matching on key variables including age, race, gender, socioeconomic status, and psychiatric diagnosis. RESULTS: Sexually abused and control group inpatient adolescents were similar on standardized measures of psychological distress and family functioning. Statistically significant differences were found between groups on substance abuse measures. CONCLUSION: The phenomenology of the relationship between sexual abuse and substance abuse and the implications for inpatient treatment are summarized. PMID- 8407770 TI - Childhood sexual abuse in anorexia nervosa and bulimia nervosa: a pilot study. AB - OBJECTIVES: The aims of this pilot study were to determine the feasibility and scientific merit of coordinating a large-scale investigation of childhood sexual abuse (CSA) in outpatient-eating disordered women and to evaluate the usefulness and potential adverse effects of a semistructured trauma interview. METHOD: The sample was randomly selected from subjects participating in a naturalistic longitudinal study of anorexia nervosa (AN) and bulimia nervosa (BN). The refusal rate was 26%. Twenty women with DSM-III-R AN/BN were assessed with the traumatic antecedents interview (TAI); a brief semistructured follow-up interview was administered over the telephone 3 to 8 weeks after the first interview. Subjects also completed the Dissociative Experiences Scale (DES). RESULTS: Thirteen subjects (65%) reported CSA, and those with more comorbidity reported CSA more frequently. Subjects with a history of CSA had significantly higher scores on the DES. Minimal adverse consequences to the TAI were noted. CONCLUSIONS: The high compliance rate, high prevalence of CSA, and high DES scores among our pilot sample support the merit of studying CSA in a larger sample. PMID- 8407771 TI - Family therapy of extrafamilial sexual abuse. AB - A significant portion of children referred for psychiatric treatment have been sexually abused. One of the most difficult symptom manifestations to treat in young children is the management of anger. In this case, a 4-year-old boy was sexually abused by persons outside the family. He showed symptoms of regressive behavior including encopresis, enuresis, difficulty sleeping, fearfulness, recurrent nightmares, and had hyperalertness and frequent outbursts of anger. Treatment initially involved group therapy, with a concurrent parents' group. After experiencing little improvement in group therapy, the youngster was put in individual play therapy and family therapy. Family therapy proved essential in creating a safe environment for the patient where he could learn to regulate his affect and process his traumatic experience successfully. PMID- 8407772 TI - Child sexual abuse revisited by the U.S. Supreme Court. AB - In deciding a landmark child sexual abuse case, the U.S. Supreme Court broke new ground in addressing the unique needs and qualities of child witnesses. The Court unanimously decided that the spontaneous statement of an abused child, made outside of a courtroom and while receiving medical treatment because of molestation, is trustworthy and may be allowed as evidence at trial. The Court curtailed the right of a defendant to go face to face against the child accuser, considering that a victimized child's statement, made while he or she is emotionally injured, has substantial value that cannot be duplicated simply by testifying later in court. PMID- 8407773 TI - Comparison of three theoretical models of substance use among urban minority high school students. AB - OBJECTIVE: This study compares the relative explanatory power of three theoretical models of substance use among urban minority high school students. METHOD: A substance use survey was administered in the spring of 1991 to a randomly selected sample of classrooms in the ninth through twelfth grades of three public academic high schools in a New York City borough. Survey participants (N = 919) were 59% black and 34% Hispanic; the mean age was 16.4 years. RESULTS: Substantial proportions of this sample of students reported experimental or occasional alcohol and cigarette use in the past year, and approximately one-tenth reported frequent use of these substances. Only small proportions of students reported past-year marijuana use, and few or no students reported past-year use of cocaine, crack or intravenous heroin. The socialization model of substance use was much more powerful than either the stress/strain or disaffiliation models in explaining past-year use of alcohol, cigarettes, and marijuana. However, certain variables derived from the stress/strain and disaffiliation models were important risk factors for the frequent use of these substances. CONCLUSIONS: These findings suggest that the further elucidation of the social influence process among adolescents, and the development, implementation, and evaluation of especially intensive programs aimed at high risk youths, should become adolescent substance use prevention research priorities. PMID- 8407774 TI - Role of mutual attachment in drug use: a longitudinal study. AB - OBJECTIVE: This study examines childhood aggression, mutual attachment and later drug use. METHOD: Data on 397 children and adolescents at three points in time were collected and analyzed. Mothers and their children were individually interviewed. RESULTS: A weak parent-child mutual attachment in girls can be viewed as a consequence of childhood aggression. Moreover, mutual parent-child attachment affects later drug use through three stabilities: (1) the stability of attachment during adolescence, (2) the stability of unconventionality during adolescence, and (3) the stability of drug use during adolescence. CONCLUSION: The multiple pathway perspective on drug use poses a number of ways in which to think about interventional approaches. First, interventions may be targeted toward those risk factors showing the strongest relations with later drug use. A second mode of orientation to intervention would deal with the amenability of the target to particular interventional agents. Interventions may be geared to intraindividual characteristics or may focus more attention on familial characteristics. A third way of considering interventions, as suggested by the developmental pathways to drug use, seeks to address the temporal order of risk factors leading to drug use. PMID- 8407775 TI - Long-term psychopathological and cognitive outcome of children with fetal alcohol syndrome. AB - OBJECTIVE: The long-term outcome of a large cohort of children suffering from fetal alcohol syndrome was studied. METHOD: Structured psychiatric interviews, behavior checklists for parents and teachers, and intelligence tests were used. Assessments took place during preschool age, early school age (6 to 12 years), and late school age (> or = 13 years). RESULTS: There was an excess of psychopathology with a wide variety of psychiatric syndromes in this cohort. Hyperkinetic disorders, emotional disorders, sleep disorders, and abnormal habits and stereotypes persisted over time. Interview findings were largely in accordance with parents' and teachers' questionnaire findings. Intelligence test findings included a large proportion of mentally retarded children and displayed high stability at follow-up. CONCLUSIONS: The development of children suffering from fetal alcohol syndrome is jeopardized by a high rate of persistent psychiatric and cognitive impairments. PMID- 8407776 TI - Psychopathology in children of alcoholics. AB - OBJECTIVE: To assess psychopathology in 125 and 158 children who are offspring of alcoholic and control parents. METHOD: Parents and children were interviewed by means of structured interviews. Parents were interviewed about themselves and about their children. Teacher reports were obtained. A total of 158 young people ages 6 to 18 years were in the study, although some of the analyses use only 125. RESULTS: Children of alcoholic parents showed higher rates of oppositional and conduct disorders but not attention deficit disorder. Children of alcoholic parents did not have significantly higher rates of depression, but they may be at risk for anxiety. These children also showed increased incidence of alcohol and other substance use but not abuse or dependence as defined by DSM-III. Few differences were detected with respect to self-esteem and achievement tests among the groups. There were no differences in the rates of psychopathology between offspring of alcoholic versus antisocial parents. CONCLUSION: These data indicate that children of alcoholics exhibit high rates of psychopathology and may be at risk specifically for oppositional and conduct disorders but not for depression. There are few differences between alcoholics and controls with respect to self esteem and achievement tests. PMID- 8407777 TI - The sigma 54 bacterial enhancer-binding protein family: mechanism of action and phylogenetic relationship of their functional domains. PMID- 8407778 TI - Degradation of chloroaromatics: purification and characterization of maleylacetate reductase from Pseudomonas sp. strain B13. AB - Maleylacetate reductase of Pseudomonas sp. strain B13 was purified to homogeneity by chromatography on DEAE-cellulose, Butyl-Sepharose, Blue-Sepharose, and Sephacryl S100. The final preparation gave a single band by polyacrylamide gel electrophoresis under denaturing conditions and a single symmetrical peak by gel filtration under nondenaturing conditions. The subunit M(r) value was 37,000 (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Estimation of the native M(r) value by gel filtration gave a value of 74,000 with a Superose 6 column, indicating that the enzyme is dimeric. The pH and temperature optima were 5.4 and 50 degrees C, respectively. The pI of the enzyme was estimated to be 7.0. The apparent Km values for maleylacetate and NADH were 58 and 30 microM, respectively, and the maximum velocity was 832 U/mg of protein for maleylacetate. Maleylacetate and various substituted maleylacetates, such as 2-chloro- and 2-methyl-maleylacetate, were reduced at significant rates. NADPH was also used as a cofactor instead of NADH with nearly the same Vmax value, but its Km value was estimated to be 77 microM. Reductase activity was inhibited by a range of thiol-blocking reagents. The absorption spectrum indicated that there was no bound cofactor or prosthetic group in the enzyme. PMID- 8407779 TI - Penicillin-binding proteins from Erwinia amylovora: mutants lacking PBP2 are avirulent. AB - Radiolabelled penicillin G was used to examine penicillin-binding proteins (PBPs) from Erwinia amylovora (OT1). This procedure identified seven PBPs with molecular masses ranging from 22 to 83 kDa. E. amylovora PBPs were compared with those from Escherichia coli (JM101) and from two spherical, avirulent TnphoA mutants derived from OT1. Radiolabelled penicillin G bound to only six proteins from the spherical mutants which lacked a 69-kDa PBP. The spherical mutants could be complemented by the cloned E. coli pbpA-rodA operon, which restored both cell shape and virulence to apple seedlings. This suggested that the E. amylovora 69 kDa PBP is probably the functional equivalent of the E. coli PBP2 protein. Southern blot analysis using the E. coli rodA and pbpA genes as radiolabelled probes showed that TnphoA had inserted into the E. amylovora equivalent of the E. coli rodA-pbpA operon. Southern blots to chromosomal DNAs of the two spherical mutants, using the cloned hrp and dsp genes from E. amylovora as radiolabelled probes, confirmed that the TnphoA insertions were not located in the region of the E. amylovora chromosome postulated to encode known virulence factors. Both of the spherical TnphoA mutants synthesized amounts of extracellular polysaccharide equivalent to those synthesized by the wild-type strain (OT1), were resistant to lysis in distilled water and to lysozyme, and elicited the hypersensitive response on nonhost plants. These results indicate a possible role for cell shape in the virulence of this plant pathogen. PMID- 8407780 TI - S-layer protein of Lactobacillus acidophilus ATCC 4356: purification, expression in Escherichia coli, and nucleotide sequence of the corresponding gene. AB - The cell surfaces of several Lactobacillus species are covered by a regular layer composed of a single species of protein, the S-protein. The 43-kDa S-protein of the neotype strain Lactobacillus acidophilus ATCC 4356, which originated from the pharynx of a human, was purified. Antibodies generated against purified S-protein were used to screen a lambda library containing chromosomal L. acidophilus ATCC 4356 DNA. Several phages showing expression of this S-protein in Escherichia coli were isolated. A 4.0-kb DNA fragment of one of those phages hybridized to a probe derived from an internal tryptic fragment of the S-protein. The slpA gene, coding for the surface layer protein, was located entirely on the 4.0-kb fragment as shown by deletion analysis. The nucleotide sequence of the slpA gene was determined and appeared to encode a protein of 444 amino acids. The first 24 amino acids resembled a putative secretion signal, giving rise to a mature S protein of 420 amino acids (44.2 kDa). The predicted isoelectric point of 9.4 is remarkably high for an S-protein but is in agreement with the data obtained during purification. The expression of the entire S-protein or of large, C terminally truncated S-proteins is unstable in E. coli. PMID- 8407781 TI - CbbR, a LysR-type transcriptional activator, is required for expression of the autotrophic CO2 fixation enzymes of Xanthobacter flavus. AB - Xanthobacter flavus is able to grow autotrophically with the enzymes of the Calvin cycle for the fixation of CO2, which are specified by the cbbLSXFP gene cluster. Previously, the 5' end of an open reading frame (cbbR), displaying a high sequence similarity to the LysR family of regulatory proteins and transcribed divergently from cbbLSXFP, was identified (W. G. Meijer, A. C. Arnberg, H. G. Enequist, P. Terpstra, M. E. Lidstrom, and L. Dijkhuizen, Mol. Gen. Genet. 225:320-330, 1991). This paper reports the complete nucleotide sequence of cbbR and a functional characterization of the gene. The cbbR gene of X. flavus specifies a 333-amino-acid polypeptide, with a molecular weight of 35,971. Downstream from cbbR, the 3' end of an open reading frame displaying a high similarity to ORF60K from Pseudomonas putida and ORF261 from Bacillus subtilis was identified. ORF60K and ORF261 are located at the replication origin of the bacterial chromosome. Inactivation of cbbR, via the insertion of an antibiotic resistance gene, rendered X. flavus unable to grow autotrophically. This was caused not by an inability to oxidize autotrophic substrates (e.g., formate) but by a complete lack of expression of the cbb genes. The expression of the CbbR protein in Escherichia coli was achieved by placing cbbR behind a strong promoter and optimization of the translational signals of cbbR. CbbR binds specifically to two binding sites in the cbbR-cbbL intergenic region. PMID- 8407782 TI - Isolation and characterization of a cyanide dihydratase from Bacillus pumilus C1. AB - A cyanide-degrading enzyme from Bacillus pumilus C1 has been purified and characterized. This enzyme consisted of three polypeptides of 45.6, 44.6, and 41.2 kDa; the molecular mass by gel filtration was 417 kDa. Electron microscopy revealed a multimeric, rod-shaped protein approximately 9 by 50 nm. Cyanide was rapidly degraded to formate and ammonia. Enzyme activity was optimal at 37 degrees C and pH 7.8 to 8.0. Activity was enhanced by Sc3+, Cr3+, Fe3+, and Tb3+; enhancement was independent of metal ion concentration at concentrations above 5 microM. Reversible enhancement of enzymatic activity by azide was maximal at 4.5 mM azide and increased with time. No activity was recorded with the cyanide substrate analogs CNO-, SCN-, CH3CN, and N3- and the possible degradation intermediate HCONH2. Kinetic studies indicated a Km of 2.56 +/- 0.48 mM for cyanide and a Vmax of 88.03 +/- 4.67 mmol of cyanide per min/mg/liter. The Km increased approximately twofold in the presence of 10 microM Cr3+ to 5.28 +/- 0.38 mM for cyanide, and the Vmax increased to 197.11 +/- 8.51 mmol of cyanide per min/mg/liter. We propose naming this enzyme cyanide dihydratase. PMID- 8407783 TI - Processing of lipid-modified prolipoprotein requires energy and sec gene products in vivo. AB - The kinetics of processing of glyceride-modified prolipoprotein that accumulated in globomycin-treated Escherichia coli has been found to be affected by sec mutations, i.e., secA, secE, secY, secD, and secF, and by metabolic poisons which affect proton motive force (PMF). The effect of sec mutations on processing of glyceride-modified prolipoprotein in vivo was not due to a secondary effect on PMF. Neither a secF mutation nor metabolic poisons affected the processing of previously accumulated proOmpA protein in vivo, suggesting that the requirements for functional sec gene products and PMF are specific to the processing of lipoprotein precursors by signal peptidase II. PMID- 8407784 TI - Suppression of recJ mutations of Escherichia coli by mutations in translation initiation factor IF3. AB - We have isolated genetic suppressors of mutations in the recJ gene of Escherichia coli in a locus we term srjA. These srjA mutations cause partial to complete alleviation of the recombination and UV repair defects conferred by recJ153 and recJ154 mutations in a recBC sbcA genetic background. The srjA gene was mapped to 37.5 min on the E. coli chromosome. This chromosomal region from the srjA5 strain was cloned into a plasmid vector and was shown to confer recJ suppression in a dominant fashion. Mutational analysis of this plasmid mapped srjA to the infC gene encoding translation initiation factor 3 (IF3). Sequence analysis revealed that all three srjA alleles cause amino acid substitutions of IF3. Suppression of recJ was shown to be allele specific: recJ153 and recJ154 mutations were suppressible, but recJ77 and the insertion allele recJ284::Tn10 were not. In addition, growth medium-conditional lethality was observed for strains carrying srjA mutations with the nonsuppressible recJ alleles. When introduced into recJ+ strains, srjA mutations conferred hyperrecombinational and hyper-UVr phenotypes. An interesting implication of these genetic properties of srjA suppression is that IF3 may regulate the expression of recJ and perhaps other recombination genes and hence may regulate the recombinational capacity of the cell. PMID- 8407785 TI - Three distinct secreted aspartyl proteinases in Candida albicans. AB - The secreted aspartyl proteinases of Candida albicans (products of the SAP genes) are thought to contribute to virulence through their effects on Candida adherence, invasion, and pathogenicity. From a single strain of C. albicans (WO 1) which expresses a phenotypic switching system, three secreted aspartyl proteinases have been identified as determined by molecular weight and N-terminal sequence. Each of the three identified proteins represents the mature form of one of three distinct proteinase isoenzymes, two of which correspond to the recently cloned SAP1 and SAP2 genes (previously referred to as CAP, PEP, or PRA). A genomic library was screened under low-stringency hybridization conditions with a polymerase chain reaction fragment from SAP1. In addition to clones of SAP1 and SAP2, a clone containing SAP3, a novel third secreted proteinase gene, was identified and sequenced. The three aspartyl proteinase isoenzymes differ in primary sequence and pI, suggesting that they may play different roles in virulence and pathogenesis. All three of these proteinases are expressed in the same strain. However, the pattern of proteinase expression is correlated with the switch phenotype of the cell. Opaque cells of strain WO-1 express Sap1 and Sap3, while white cells of the same strain express Sap2. The differential expression of three Sap proteinases may contribute to virulence in C. albicans. PMID- 8407786 TI - Tetracycline regulation of genes on Bacteroides conjugative transposons. AB - Human colonic Bacteroides species harbor a family of large conjugative transposons, called tetracycline resistance (Tcr) elements. Activities of these elements are enhanced by pregrowth of bacteria in medium containing tetracycline, indicating that at least some Tcr element genes are regulated by tetracycline. Previously, we identified a central regulatory locus on the Tcr elements that contained two genes, rteA and rteB, which appeared to encode a two-component regulatory system (A. M. Stevens, J. M. Sanders, N. B. Shoemaker, and A. A. Salyers, J. Bacteriol. 174:2935-2942, 1992). In the present study, we describe a gene which is located downstream of rteB in a separate transcriptional unit and which requires RteB for expression. Sequence analysis of this gene showed that it encoded a 217-amino-acid protein, which had no significant sequence similarity to any proteins in the GenBank or EMBL data base. An insertional disruption in the gene abolished self-transfer of the Tcr element to Bacteroides recipients, indicating that the gene was essential for self-transfer. The disruption also affected mobilization of coresident plasmids. Mobilization frequency was reduced 100- to 1,000-fold if the recipient was Escherichia coli but was not affected to the same extent if the recipient was an isogenic Bacteroides strain. The complex phenotype of the disruption mutant suggested that the newly identified gene, like rteA and rteB, had a regulatory function. Accordingly, it has been designated rteC. Our results indicate that regulation of Tc(r) element functions is unexpectedly complex and may involve a cascade of regulators, with RteA and RteB exerting central control over secondary regulators like RteC, which in turn control subsets of Tcr element structural genes. PMID- 8407787 TI - Identification of the glmU gene encoding N-acetylglucosamine-1-phosphate uridyltransferase in Escherichia coli. AB - The physiological properties of the EcoURF-1 open reading frame, which precedes the glmS gene at 84 min on the Escherichia coli chromosome (J. E. Walker, N. J. Gay, M. Saraste, and A. N. Eberle, Biochem. J. 224:799-815, 1984), were investigated. A thermosensitive conditional mutant in which the synthesis of the gene product was impaired at 43 degrees C was constructed. The inactivation of the gene in exponentially growing cells rapidly inhibited peptidoglycan synthesis. As a result, various alterations of cell shape were observed, and cell lysis finally occurred when the peptidoglycan content was 37% lower than that of normally growing cells. Analysis of the pools of peptidoglycan precursors revealed a large accumulation of N-acetylglucosamine-1-phosphate and the concomitant depletion of the pools of the seven peptidoglycan nucleotide precursors located downstream in the pathway, a result indicating that the mutational block was in the step leading from N-acetylglucosamine-1-phosphate and UTP to the formation of UDP-N-acetylglucosamine. In vitro assays showed that the overexpression of this gene in E. coli cells, directed by appropriate plasmids, led to a high overproduction (from 25- to 410-fold) of N-acetylglucosamine-1 phosphate uridyltransferase activity. This allowed us to purify this enzyme to homogeneity in only two chromatographic steps. The gene for this enzyme, which is essential for peptidoglycan and lipopolysaccharide biosyntheses, was designated glmU. PMID- 8407788 TI - Molecular characterization of the Enterobacter aerogenes tonB gene: identification of a novel type of tonB box suppressor mutant. AB - The tonB gene of Enterobacter aerogenes was cloned, sequenced, and expressed in Escherichia coli. It complemented an E. coli tonB mutant as efficiently as E. coli tonB, except for colicin B and D sensitivities. However, colicin B and D sensitivities were complemented by a derivative in which the aspartate at position 165 was replaced by a glutamine (TonBD-165-->Q) by site-directed mutagenesis. In E. coli, the corresponding amino acid is a glutamine (Q-160) which is known to be altered in most mutants showing suppression of the btuB451 mutation. Fourteen independent btuB451 suppressor mutations in E. aerogenes tonB which all had suffered the same point mutation resulting in a change from glycine to valine at position 239 (G-239-->V) of the C-terminal end of the protein were isolated. The mutation was located within a region which is nonessential for function of E. aerogenes TonB as well as E. coli TonB. A constructed double mutation, expressing a D-165-->Q/G-239-->V derivative, no longer acted as a btuB451 suppressor. However, it restored colicin B and D sensitivities even more efficiently than the D-165-->Q derivative. Corresponding mutations constructed in E. coli tonB, giving rise to Q-160-->D, G-234-->V, and Q-160-->D/G-234-->V derivatives, showed phenotypes comparable to the E. aerogenes mutations. We take this as evidence that at least a functional interaction between the D-165 (Q-160 in E. coli) and the G-239 (G-234 in E. coli) region is necessary for TonB function. The implications of this interaction for functional instability of TonB are discussed. PMID- 8407789 TI - vsrB, a regulator of virulence genes of Pseudomonas solanacearum, is homologous to sensors of the two-component regulator family. AB - Pseudomonas solanacearum, an important wilt pathogen of many plants, produces several extracellular proteins (EXPs) and extracellular polysaccharides (EPSs) that contribute to its virulence. Using TnphoA mutagenesis, we discovered a new gene, vsrB, that when inactivated causes a major reduction in the virulence and production of an EPS. Analysis of eps::lacZ reporters showed that vsrB is required for maximal expression (transcription) of eps, whose products are required for production of EPS I, a major virulence determinant. Analysis of EXPs in culture supernatants revealed that inactivation of vsrB also causes reduced production of two major EXPs, with molecular masses of 28 and 97 kDa, and a simultaneous 15-fold increase in levels of another EXP, PglA endopolygalacturonase. The vsrB gene was cloned from a P. solanacearum genomic library by complementation of the nonmucoid phenotype of the vsrB::TnphoA mutant and then subcloned on a 2.4-kb DNA fragment. TnphoA fusion analysis and subcellular localization of the vsrB gene product in Escherichia coli maxicells suggest that it is a ca. 60-kDa transmembrane protein. The nucleotide sequence of the 2.4-kb DNA fragment was determined, and a 638-amino-acid open reading frame was found for VsrB. A search of the GenBank data base found that the central part of VsrB has homology with the histidine kinase domain of sensors in the two component regulator family, while the C terminus has homology with the phosphate receiver domain of response regulators in the same family. Genetic analysis suggests that the receiver domain is not required for vsrB function. PMID- 8407790 TI - Functional domains of S-type pyocins deduced from chimeric molecules. AB - Functional domain structures of pyocins AP41, S1, and S2 were assigned by examining the functions of chimeric pyocins and deletion derivatives. Pyocins AP41, S1, and S2 are essentially composed of three domains, the receptor-binding domain, the translocation domain, and the DNase domain, in that order from the N terminus to the C terminus. The alignment of these domains is distinct from that in E2-group colicins with functions similar to those of these pyocins. Pyocins AP41 and S2 have a fourth domain between the receptor-binding and the translocation domains, which is dispensable for their killing functions. PMID- 8407791 TI - Cloning, sequencing, and expression of the Pseudomonas putida protocatechuate 3,4 dioxygenase genes. AB - The genes that encode the alpha and beta subunits of protocatechuate 3,4 dioxygenase (3,4-PCD [EC 1.13.11.3]) were cloned from a Pseudomonas putida (formerly P. aeruginosa) (ATCC 23975) genomic library prepared in lambda phage. Plaques were screened by hybridization with degenerate oligonucleotides designed using known amino acid sequences. A 1.5-kb SmaI fragment from a 15-kb primary clone was subcloned, sequenced, and shown to contain two successive open reading frames, designated pcaH and pcaG, corresponding to the beta and alpha subunits, respectively, of 3,4-PCD. The amino acid sequences deduced from pcaHG matched the chemically determined sequence of 3,4-PCD in all except three positions. Cloning of pcaHG into broad-host-range expression vector pKMY319 allowed high levels of expression in P. putida strains, as well as in Proteus mirabilis after specific induction of the plasmid-encoded nahG promoter with salicylate. The recombinant enzyme was purified and crystallized from P. mirabilis, which lacks an endogenous 3,4-PCD. The physical, spectroscopic, and kinetic properties of the recombinant enzyme were indistinguishable from those of the wild-type enzyme. Moreover, the same transient enzyme intermediates were formed during the catalytic cycle. These studies establish the methodology which will allow mechanistic investigations to be pursued through site-directed mutagenesis of P. putida 3,4-PCD, the only aromatic ring-cleaving dioxygenase for which the three-dimensional structure is known. PMID- 8407792 TI - Isolation and characterization of Bacillus subtilis genes involved in siderophore biosynthesis: relationship between B. subtilis sfpo and Escherichia coli entD genes. AB - In response to iron deprivation, Bacillus subtilis secretes a catecholic siderophore, 2,3-dihydroxybenzoyl glycine, which is similar to the precursor of the Escherichia coli siderophore enterobactin. We isolated two sets of B. subtilis DNA sequences that complemented the mutations of several E. coli siderophore-deficient (ent) mutants with defective enterobactin biosynthesis enzymes. One set contained DNA sequences that complemented only an entD mutation. The second set contained DNA sequences that complemented various combinations of entB, entE, entC, and entA mutations. The two sets of DNA sequences did not appear to overlap. AB. subtilis mutant containing an insertion in the region of the entD homolog grew much more poorly in low-iron medium and with markedly different kinetics. These data indicate that (i) at least five of the siderophore biosynthesis genes of B. subtilis can function in E. coli, (ii) the genetic organization of these siderophore genes in B. subtilis is similar to that in E. coli, and (iii) the B. subtilis entD homolog is required for efficient growth in low-iron medium. The nucleotide sequence of the B. subtilis DNA contained in plasmid pENTA22, a clone expressing the B. subtilis entD homolog, revealed the presence of at least two genes. One gene was identified as sfpo, a previously reported gene involved in the production of surfactin in B. subtilis and which is highly homologous to the E. coli entD gene. We present evidence that the E. coli entD and B. subtilis sfpo genes are interchangeable and that their products are members of a new family of proteins which function in the secretion of peptide molecules. PMID- 8407793 TI - Characterization of the ferrous iron uptake system of Escherichia coli. AB - Escherichia coli has an iron(II) transport system (feo) which may make an important contribution to the iron supply of the cell under anaerobic conditions. Cloning and sequencing of the iron(II) transport genes revealed an open reading frame (feoA) possibly coding for a small protein with 75 amino acids and a membrane protein with 773 amino acids (feoB). The upstream region of feoAB contained a binding site for the regulatory protein Fur, which acts with iron(II) as a corepressor in all known iron transport systems of E. coli. In addition, a Fnr binding site was identified in the promoter region. The FeoB protein had an apparent molecular mass of 70 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was localized in the cytoplasmic membrane. The sequence revealed regions of homology to ATPases, which indicates that ferrous iron uptake may be ATP driven. FeoA or FeoB mutants could be complemented by clones with the feoA or feoB gene, respectively. PMID- 8407794 TI - Molecular characterization of a STreptococcus mutans mutant altered in environmental stress responses. AB - A mutant defective in aciduricity, GS5Tn1, was constructed following mutagenesis of Streptococcus mutans GS5 with the conjugative transposon Tn916. The mutant grew poorly at acidic pH levels and was sensitive to high osmolarity and elevated temperatures. These properties resulted from a single insertion of Tn916 into the GS5 chromosome, and the DNA fragment harboring the transposon was isolated into the cosmid vector, charomid 9-20. Spontaneous excision of Tn916 from the cosmid revealed that Tn916 inserted into a 8.6-kb EcoRI fragment. On the basis of the restriction analyses of insert fragments, it was found that Tn916 inserted into a 0.9-kb EcoRI-XbaI fragment. Nucleotide sequence analysis of this fragment indicated the presence of two open reading frames, ORF1 and ORF2. By using a marker rescue strategy, a 6.0-kb HindIII fragment including the target site for Tn916 insertion and the 5' end of ORF1 was isolated and sequenced. The deduced amino acid sequences of ORF1 and ORF2 showed significant homology with the diacylglycerol kinase and Era proteins, respectively, from Escherichia coli. Nucleotide sequence analysis of the Tn916 insertion junction region in the GS5Tn1 chromosome revealed that the transposon inserted near the 3' terminus of ORF1. Restoration of ORF1 to its original sequence in mutant GS5Tn1 was carried out following transformation with integration vector pVA891 containing an intact ORF1. The resultant transformant showed wild-type levels of aciduricity as well as resistance to elevated temperatures and high osmolarity. These results suggest that the S. mutans homolog of diacylglycerol kinase is important for adaptation of the organism to several environmental stress signals. PMID- 8407795 TI - High-resolution visualization by field emission scanning electron microscopy of Enterococcus faecalis surface proteins encoded by the pheromone-inducible conjugative plasmid pCF10. AB - Enterococcus faecalis can acquire antibiotic resistance and virulence genes by transfer of pheromone-inducible conjugative plasmids such as pCF10, which encodes tetracycline resistance. Two pCF10-encoded cell surface proteins, Sec10 and Asc10, have been previously shown to play an important role in the transfer of this plasmid. We used high-resolution, field emission scanning electron microscopy to visualize these proteins on the surfaces of a series of isogenic strains of E. faecalis. Immunogold labeling, using both 6- and 12-nm colloidal gold, unambiguously demonstrated the expression and distribution of Sec10 and Asc10 on the surface of the E. faecalis cells. On unlabeled E. faecalis cells which expressed either Sec10 or Asc10, the former appeared to be more readily detected. Immunogold labeling of E. faecalis cells expressing both Asc10 and Sec10 clearly demonstrated the abundance and intermixing of both proteins on the cell surface except at septal regions. Sec10 was observed to be distributed over the cell surface. At regions of cell-cell contact, fine strands representing Asc10 were observed directly attaching adjacent cells to one another. PMID- 8407796 TI - Multiple factors underlying the maximum motility of Escherichia coli as cultures enter post-exponential growth. AB - Motility and chemotaxis allow cells to move away from stressful microenvironments. Motility of Escherichia coli in batch cultures, as measured by cell swimming speed, was low in early-exponential-phase cells, peaked as the cells entered post-exponential phase, and declined into early stationary phase. Transcription from the flhB operon and synthesis of flagellin protein similarly peaked in late exponential and early post-exponential phases, respectively. The increase in swimming speed between early-exponential and post-exponential phases was correlated with twofold increases in both flagellar length and flagellar density per cell volume. This increased investment in flagella probably reflects the increased adaptive value of motility in less favorable environments. The decrease in speed between post-exponential and stationary phases was correlated with a threefold decrease in torque produced by the flagellar motors and presumably reflects decreased proton motive force available to stationary-phase cells. PMID- 8407797 TI - Cloning and sequencing of the gene coding for the large subunit of methylamine dehydrogenase from Thiobacillus versutus. AB - The gene that codes for the alpha-subunit of methylamine dehydrogenase from Thiobacillus versutus, madA, was cloned and sequenced. It codes for a protein of 395 amino acids preceded by a leader sequence of 31 amino acids. The derived amino acid sequence was confirmed by partial amino acid sequencing. The start of the mature protein could not be determined by direct sequencing, since the N terminus appeared to be blocked. Instead, it was determined by electrospray mass spectrometry. Confirmation of the results was obtained by sequencing the N terminus after pyroglutamate aminopeptidase digestion. The sequence is homologous to the Paracoccus denitrificans nucleotide sequence. A second open reading frame, called open reading frame 3, is located immediately downstream of madA. PMID- 8407798 TI - Molecular cloning of a sporulation-specific cell wall hydrolase gene of Bacillus subtilis. AB - Southern hybridization analysis of Bacillus subtilis 168S chromosomal DNA with a Bacillus licheniformis cell wall hydrolase gene, cwlM, as a probe indicated the presence of a cwlM homolog in B. subtilis. DNA sequencing of the cwlM homologous region showed that a gene encoding a polypeptide of 255 amino acids with a molecular mass of 27,146 Da is located 625 bp upstream and in the opposite direction of spoVJ. The deduced amino acid sequence of this gene (tentatively designated as cwlC) showed an overall identity of 73% with that of cwlM and of 40% with the C-terminal half of the B. subtilis vegetative autolysin, CwlB. The construction of an in-frame cwlC-lacZ fusion gene in the B. subtilis chromosome indicated that cwlC is induced at 6 to 7 h after sporulation (t6 to t7). The spoIIIC (sigma K) mutation and earlier sporulation mutations greatly reduced the expression of the cwlC-lacZ fusion gene. Northern hybridization analysis using oligonucleotide probes of the cwlC region indicated that a unique cwlC transcript appeared at t7.5 and t9. Transcriptional start points determined by primer extension analysis suggested that the -10 region is very similar to the consensus sequence for the sigma K-dependent promoter. Insertional inactivation of the cwlC gene in the B. subtilis chromosome caused the disappearance of a 31-kDa protein lytic for Micrococcus cell walls, which is mainly located within the cytoplasmic and membrane fractions of cells at t9. The CwlC protein hydrolyzed both B. subtilis vegetative cell walls and spore peptidoglycan. PMID- 8407799 TI - Lactose permease mutants which transport (malto)-oligosaccharides. AB - Lactose permease mutants, which were previously isolated in sugar specificity studies, were screened for their abilities to transport the trisaccharide maltotriose. Six multiple mutants (e.g., five double mutants and one triple mutant) were identified as forming fermentation-positive colonies on maltotriose MacConkey plates and were also shown to grow on maltotriose minimal plates. All of these multiple mutants contained a combination of two or three amino acid substitutions at position 177, 236, 306, or 322 within the permease. In contrast, none of the corresponding single mutants at these locations were observed to exhibit an enhanced rate of maltotriose transport. In whole-cell assays, the multiple mutants were shown to transport relatively long alpha nitrophenylglucoside (alpha NPG) molecules. In certain cases, alpha NPG molecules containing up to four glucose residues in addition to the nitrophenyl group were shown to be transported to a significant degree. Overall, the abilities of lactose permease mutants to transport maltotriose and long alpha NPGs are discussed with regard to the dimensions of the sugar and the mechanism of sugar transport. PMID- 8407800 TI - Characterization of genes for an alternative nitrogenase in the cyanobacterium Anabaena variabilis. AB - Anabaena variabilis ATCC 29413 is a heterotrophic, nitrogen-fixing cyanobacterium that has been reported to fix nitrogen and reduce acetylene to ethane in the absence of molybdenum. DNA from this strain hybridized well at low stringency to the nitrogenase 2 (vnfDGK) genes of Azotobacter vinelandii. The hybridizing region was cloned from a lambda EMBL3 genomic library of A. variabilis, mapped, and sequenced. The deduced amino acid sequences of the vnfD and vnfK genes of A. variabilis showed only about 56% similarity to the nifDK genes of Anabaena sp. strain PCC 7120 but were 76 to 86% similar to the anfDK or vnfDK genes of A. vinelandii. The organization of the vnf gene cluster in A. variabilis was similar to that of A. vinelandii. However, in A. variabilis, the vnfG gene was fused to vnfD; hence, this gene is designated vnfDG. A vnfH gene was not contiguous with the vnfDG gene and has not yet been identified. A mutant strain, in which a neomycin resistance cassette was inserted into the vnf cluster, grew well in a medium lacking a source of fixed nitrogen in the presence of molybdenum but grew poorly when vanadium replaced molybdenum. In contrast, the parent strain grew equally well in media containing either molybdenum or vanadium. The vnf genes were transcribed in the absence of molybdenum, with or without vanadium. The vnf gene cluster did not hybridize to chromosomal DNA from Anabaena sp. strain PCC 7120 or from the heterotrophic strains, Nostoc sp. strain Mac and Nostoc sp. strain ATCC 29150. A hybridizing ClaI fragment very similar in size to the A. variabilis ClaI fragment was present in DNA isolated from several independent, cultured isolates of Anabaena sp. from the Azolla symbiosis. PMID- 8407801 TI - moaR, a gene that encodes a positive regulator of the monoamine regulon in Klebsiella aerogenes. AB - We cloned and sequenced a Klebsiella aerogenes gene (moaR) for activation of arylsulfatase synthesis by tyramine. This gene was cloned by complementation of a K. aerogenes mutant in which tyramine fails to relieve the arylsulfatase repression caused by sulfur compounds. The moaR gene also activated induction of the synthesis of both tyramine oxidase and the 30-kDa protein that is specifically induced by high concentrations of tyramine or catecholamines. The moaR gene on the chromosome of the wild-type strain of K. aerogenes was disrupted by homologous recombination with a plasmid containing the inactivated moaR. The resultant mutant showed the same phenotype as previously isolated atsT mutant strains that are negative for the derepressed synthesis of arylsulfatase. In this mutant strain, tyramine also failed to induce the synthesis of tyramine oxidase or the production of a 30-kDa protein. The moaR gene is capable of encoding a protein of 26,238 Da. The putative MoaR protein has a helix-turn-helix motif in its C terminus. Thus, it seems likely that the MoaR protein regulates the operons by binding to the regulatory region of the monoamine regulon. The MoaR protein is subject to autogenous control, which was shown by use of a moaR'-lacZ transcriptional fusion. PMID- 8407802 TI - Molecular cloning and characterization of acrA and acrE genes of Escherichia coli. AB - The DNA fragment containing the acrA locus of the Escherichia coli chromosome has been cloned by using a complementation test. The nucleotide sequence indicates the presence of two open reading frames (ORFs). Sequence analysis suggests that the first ORF encodes a 397-residue lipoprotein with a 24-amino-acid signal peptide at its N terminus. One inactive allele of acrA from strain N43 was shown to contain an IS2 element inserted into this ORF. Therefore, this ORF was designated acrA. The second downstream ORF is predicted to encode a transmembrane protein of 1,049 amino acids and is named acrE. Genes acrA and acrE are probably located on the same operon, and both of their products are likely to affect drug susceptibilities observed in wild-type cells. The cellular localizations of these polypeptides have been analyzed by making acrA::TnphoA and acrE::TnphoA fusion proteins. Interestingly, AcrA and AcrE share 65 and 77% amino acid identity with two other E. coli polypeptides, EnvC and EnvD, respectively. Drug susceptibilities in one acrA mutant (N43) and one envCD mutant (PM61) have been determined and compared. Finally, the possible functions of these proteins are discussed. PMID- 8407803 TI - D-arabitol metabolism in Candida albicans: studies of the biosynthetic pathway and the gene that encodes NAD-dependent D-arabitol dehydrogenase. AB - Candida albicans produces large amounts of the pentitol D-arabitol in culture and in infected mammalian hosts, but the functional and pathogenic significance of D arabitol in C. albicans is not known. In this study, we sought to elucidate the pathway by which C. albicans synthesizes D-arabitol and to identify and characterize key enzymes in this pathway. C. albicans B311 produced D-[14C 1]arabitol from [14C-2]glucose; this finding implies on structural grounds that D ribulose-5-PO4 from the pentose pathway is the major metabolic precursor of D arabitol. NAD- or NADP-dependent pentitol dehydrogenases catalyze the final steps in D-arabitol biosynthesis in other fungi; therefore, lysates of C. albicans B311 were tested for enzymes of this class and were found to contain a previously unknown NAD-dependent D-arabitol dehydrogenase (ArDH). The ArDH structural gene was cloned by constructing a new D-arabitol utilization pathway in Escherichia coli. The C. albicans ArDH gene expressed in E. coli and Saccharomyces cerevisiae an enzyme that catalyzes the reaction D-arabitol + NAD <-->D-ribulose + NADH; this gene was present as a single copy per haploid genome, and its deduced peptide sequence was homologous with sequences of several members of the short chain dehydrogenase family of enzymes. These results suggest that (i) C. albicans synthesizes D-arabitol by dephosphorylating and reducing the pentose pathway intermediate D-ribulose-5-PO4 and (ii) ArDH catalyzes the final step in this pathway. PMID- 8407804 TI - Characterization of a new sporulation factor in Bacillus subtilis. AB - We report the existence and partial purification of sporulation factor, which stimulates sporulation of Bacillus subtilis at low cell density. Proline or arginine is required for stimulation under the conditions of our assay. Sporulation factor is a small heat-stable substance produced by the cells during exponential growth phase. It is required in small amounts and is resistant to various proteolytic agents. Several spo mutants were tested for the ability to produce functional sporulation factor. All of these mutants produce factor and do not sporulate in the presence of factor from wild-type cells. Sporulation factor is not involved in the induction of alpha-amylase synthesis at the initiation of sporulation. PMID- 8407805 TI - cobU-dependent assimilation of nonadenosylated cobinamide in cobA mutants of Salmonella typhimurium. AB - The cobA locus of Salmonella typhimurium is involved in the assimilation of nonadenosylated cobinamide, (CN)2CBI, into cobalamin (CBL) under aerobic and anaerobic growth conditions. Aerobically, cobA mutants are unable to assimilate (CN)2CBI into CBL. However, under anaerobic conditions, cobA mutants assimilate (CN)2CBI into CBL as efficiently as cobA+ strains. On the basis of this observation, we postulated the existence of a cobA-independent pathway for the assimilation of (CN)2CBI into CBL that is functional under anaerobic growth conditions (J. C. Escalante-Semerena, S.-J. Suh, and J. R. Roth, J. Bacteriol. 172:273-280, 1990). In this paper, we report the isolation and initial genetic characterization of derivatives of cobA mutants that are unable to assimilate (CN)2CBI into CBL during anaerobic growth. As demonstrated by complementation analysis, marker rescue, and DNA sequencing data, these mutations are alleles of cobU, a gene involved in the assembly of the nucleotide loop of CBL. We have shown that the block in CBL synthesis in these cobU cobA double mutant strains can be corrected by exogenous adenosyl-CBI. Our data indicate that this new class of cobU mutations blocks CBL biosynthesis but does not destroy the putative kinase-guanylyltransferase activities of the CobU protein. We propose that this new class of cobU mutations may affect an as yet unidentified ATP:corrinoid adenosyltransferase activity of the CobU protein. Alternatively, such mutations may alter the ability of CobU to use nonadenosylated CBI as a substrate. PMID- 8407806 TI - Cloning and nucleotide sequence of a plasmid-carried gene coding for a minor small, acid-soluble protein from Bacillus megaterium spores. AB - The gene (termed sspG) coding for a small, acid-soluble protein (SASP) from spores of Bacillus megaterium QMB1551, termed SASP-G, has been cloned, and its nucleotide sequence has been determined. SASP-G is a 42-residue protein containing 2 tryptophan and 11 lysine residues, including a hexalysine sequence, and is not homologous to any previously described SASP. The sspG gene appears to be an additional member of the sigma G regulon. No gene homologous to sspG is present in B. cereus T or B. subtilis 168. The reason for the absence of sspG from other Bacillus species appears to be that in B. megaterium, sspG is present only on a 111-kb plasmid; this plasmid is not present in B. cereus T or B. subtilis 168. The sspG gene is the first forespore-expressed gene found to be on a plasmid. PMID- 8407807 TI - Evidence that the Rad1 and Rad10 proteins of Saccharomyces cerevisiae participate as a complex in nucleotide excision repair of UV radiation damage. AB - A newly characterized rad1 missense mutation (rad1-20) in the yeast Saccharomyces cerevisiae maps to a region of the Rad1 polypeptide known to be required for Rad1 Rad10 complex formation. The UV sensitivity of the rad1-20 mutant can be partially and specifically corrected by overexpression of wild-type Rad10 protein. These results suggest that complex formation between the Rad1 and Rad10 proteins is required for nucleotide excision repair. PMID- 8407808 TI - The degA gene product accelerates degradation of Bacillus subtilis phosphoribosylpyrophosphate amidotransferase in Escherichia coli. AB - A search for genes involved in the inactivation and degradation of enzymes in sporulating Bacillus subtilis led to identification of the B. subtilis degA gene, whose product stimulates degradation of B. subtilis glutamine phosphoribosylpyrophosphate amidotransferase in Escherichia coli cells. degA encodes a 36.7-kDa protein that has sequence similarity to several E. coli and B. subtilis regulatory proteins of the LacI class. B. subtilis degA::cat insertional inactivation mutants had no detectable defect in the inactivation or degradation of phosphoribosylpyrophosphate amidotransferase in glucose- or lysine-starved B. subtilis cells, however. We suggest that degA encodes either a novel protease or, more likely, a gene that stimulates production of such a protease. PMID- 8407809 TI - Construction of recombination-deficient strains of Streptococcus gordonii by disruption of the recA gene. AB - Degenerate oligonucleotide primers were used in a polymerase chain reaction (PCR) to amplify a region of the recA sequence of Streptococcus gordonii Challis. The resulting PCR fragment was cloned into the suicide vector pAM6199 and introduced into strain Challis, giving rise to recombination-deficient strains in which the recA gene was specifically inactivated. PMID- 8407810 TI - A growth rate-limiting process in the last growth phase of the yeast life cycle involves RPB4, a subunit of RNA polymerase II. AB - Saccharomyces cerevisiae cells, grown on a fermentable carbon source, display two growth phases before they enter the stationary phase: a rapid phase (log phase) followed by a slow phase. It was previously shown that a subunit of the yeast RNA polymerase II, RPB4, positively affects the activity of the enzyme in post-log phases but has little or no effect on its activity in log phase. Here, I show that RPB4 level limits the growth rate during the slow growth phase. Thus, a small increase in RPB4 protein level, in cells carrying multiple copies of the RPB4 gene, results in an almost twofold increase in the growth rate during this phase. Furthermore, RPB4 expression is differentially regulated in the two growth phases. During the slow growth phase, a posttranscriptional process which controls the RPB4 level and thus can control growth rate becomes active. These results reveal a complex growth control mechanism, in which the transcriptional apparatus is probably a limiting element, operating in the last stages of the yeast growth. PMID- 8407811 TI - Identification of a purC gene from Streptococcus pneumoniae. AB - A gene encoding 5'-phosphoribosyl-5-aminoimidazole-4-N-succinocarboxamide synthetase was identified in Streptococcus pneumoniae as a 708-bp segment of the genome encoding a 27,001-Da protein with strong similarity to known PurC proteins. The S. pneumoniae purC gene, found immediately adjacent to the competence induction genes, comAB, was cloned and sequenced. The predicted protein product of purC displayed substantial (> 40%) identity to the entire sequence of the PurC proteins of Bacillus subtilis and Escherichia coli. Function of the S. pneumoniae gene product was demonstrated by complementation of E. coli purC mutations. PMID- 8407812 TI - Cloning of the phs genetic locus from Salmonella typhimurium and a role for a phs product in its own induction. AB - The Salmonella typhimurium phs chromosomal locus essential for the reduction of thiosulfate to hydrogen sulfide was cloned, and some features of its regulation were examined. The phs locus conferred H2S production on Escherichia coli, suggesting that it contains the structural gene for thiosulfate reductase. H2S production by the E. coli host was, as in S. typhimurium, suppressed by nitrate or glucose in the growth medium. The presence of plasmid-borne phs genes in a S. typhimurium chl+ host containing a chromosomal phs::lacZ operon fusion was found to significantly increase the relative induction efficiency of beta-galactosidase by thiosulfate. These results are consistent with a model for phs regulation in which the true inducer is not thiosulfate per se and in which the action of a phs encoded molybdoprotein, possibly the reductase itself, converts thiosulfate into a compound that resembles the true inducer more closely than does thiosulfate. PMID- 8407813 TI - Mutations in the tyrR gene of Escherichia coli which affect TyrR-mediated activation but not TyrR-mediated repression. AB - Site-directed mutagenesis has been used to further characterize amino acid residues necessary for the activation of gene expression by the TyrR protein. Amino acid substitutions have been made at positions 2, 4, 5, 6, 7, 8, 9, 10, and 16. TyrR mutants with amino acid substitutions V-5-->P (VP5), VF5, CS7, CR7, DR9, RI10, RS10, and ER16 show no or very little activation of expression of either mtr or tyrP. In each case, however, the ability to repress aroF is unaltered. Amino acid substitutions at positions 4, 6, and 8 have no effect on activation. Small internal deletions of residues 10 to 19, 20 to 29, or 30 to 39 also destroy phenylalanine- or tyrosine-mediated activation of mtr and tyrP. In these mutants repression of aroF is also unaltered. In activation-defective tyrR mutants, expression of mtr is repressed in the presence of tyrosine. This tyrosine mediated repression is trpR dependent and implies an interaction between TrpR and TyrR proteins in the presence of tyrosine. PMID- 8407814 TI - The cell-bag of enzymes or network of channels? PMID- 8407815 TI - Oxidation of D-lactate and L-lactate by Neisseria meningitidis: purification and cloning of meningococcal D-lactate dehydrogenase. AB - Neisseria meningitidis was found to contain at least two lactate-oxidizing enzymes. One of these was purified 460-fold from spheroplast membranes and found to be specific primarily for D-lactate, with low-affinity activity for L-lactate. The gene for this enzyme (dld) was cloned, and a dld mutant was constructed by insertional inactivation of the gene. The mutant was unable to grow on D-lactate but retained the ability to grow on L-lactate, providing evidence for a second lactate-oxidizing enzyme with specificity for L-lactate. High-affinity L-lactate oxidizing activity was detected in intact bacteria of both the dld+ and dld mutant strains. This L-lactate-oxidizing activity was also seen in sonicated bacteria but was reduced substantially on detergent solubilization or on preparation of spheroplast membranes. PMID- 8407816 TI - Identification of cis-acting regulatory regions upstream of the rRNA operons of Rhodobacter sphaeroides. AB - The promoter region(s) for the rRNA operons of Rhodobacter sphaeroides was identified. By utilizing both in vivo and in vitro techniques, the transcriptional start sites of all three operons were identified. Upstream of the transcriptional start, -10 and -35 promoter regions that bear little similarity to typical Escherichia coli promoter sequences were identified. In addition to the promoter sequences, probable Fis protein-binding sites were identified upstream of all three rRNA operons. Transcriptional fusions of the promoter regions from rrnA and rrnB were constructed by utilizing the reporter molecule encoded by xylE and analyzed under various growth conditions, in both a wild-type background and an rrnBC mutant background. Production of the xylE gene product (catechol 2,3-dioxygenase) was always greatest under photosynthetic growth conditions. However, the upstream region of rrnB, when fused with xylE, produced significantly more catechol 2,3-dioxygenase than did analogous regions of rrnA, suggesting that the promoters of the rrn operons differ in strength. These results were further confirmed by the study of mutant strains altered for the rrn operons either singly or in combination. Little or no expression of the xylE gene was manifest in E. coli when directed by rDNA sequences derived from R. sphaeroides. PMID- 8407817 TI - The role of single-mutant intermediates in the generation of trpAB double revertants during prolonged selection. AB - Selection-induced mutations are nonrandom mutations that occur as specific, direct responses to environmental challenges and that occur more often when they are selectively advantageous than when they are selectively neutral. One of the most puzzling examples of selection-induced mutations involved the simultaneous reversions of two mutations, one in trpA and the other in trpB, at rates that were several orders of magnitude greater than would have been predicted if the two mutations had occurred as independent events (B. G. Hall, Proc. Natl. Acad. Sci. USA 88:5882-5886, 1991). Here I examine the possibility that the double mutations might be accounted for by sequential mutations with intervening growth. PMID- 8407818 TI - The mating pair formation system of plasmid RP4 defined by RSF1010 mobilization and donor-specific phage propagation. AB - Transfer functions of the conjugative plasmid RP4 (IncP alpha) are distributed among distinct regions of the genome, designated Tra1 and Tra2. By deletion analyses, we determined the limits of the Tra1 region, essential for intraspecific Escherichia coli matings. The Tra1 core region encompasses approximately 5.8 kb, including the genes traF, -G, -H, -I, -J, and -K as well as the origin of transfer. The traM gene product, however, is not absolutely required for conjugation but significantly increases transfer efficiency. To determine the transfer phenotype of genes encoded by the Tra2 core region, we generated a series of defined Tra2 mutants. This revealed that at least trbB, -C, -E, -G, and -L are essential for RP4 conjugation. To classify these transfer functions as components of the DNA transfer and replication (Dtr) or of the mating pair formation (Mpf) system, we analyzed the corresponding derivatives with respect to mobilization of IncQ plasmids and donor-specific phage propagation. We found that all of the Tra2 genes listed above and the traG and traF genes of Tra1 are required for RSF1010 mobilization. Expression of traF from Tra1 in conjunction with the Tra2 core was sufficient for phage propagation. This implies that the TraG protein is not directly involved in pilus formation and potentially connects the relaxosome with proteins enabling the membrane passage of the DNA. The proposed roles of the RP4 transfer gene products are discussed in the context of virulence functions encoded by the evolutionarily related Ti T-DNA transfer system of agrobacteria. PMID- 8407819 TI - Molecular cloning and nucleotide sequence of a putative trpDC(F)BA operon in Buchnera aphidicola (endosymbiont of the aphid Schizaphis graminum). AB - A 8,392-nucleotide-long DNA fragment from Buchnera aphidicola (endosymbiont of the aphid Schizaphis graminum) contained five genes of the tryptophan biosynthetic pathway [trpDC(F)BA] which code for enzymes converting anthranilate to tryptophan. These genes are probably arranged as a single transcription unit. Downstream of the trp genes were ORF-V, ORF-VI, and P14, three open reading frames which in Escherichia coli are also found downstream of the trp operon. Upstream of the B. aphidicola trp genes were two unidentified open reading frames, one of which potentially codes for a membrane-spanning protein with a leader sequence. Evidence for the presence of trpB in the endosymbionts of eight additional species of aphids and two species of whiteflies was obtained. These results as well as those of A. E. Douglas and W. A. Prosser (J. Insect Physiol. 38:565-568, 1992) suggest that aphid endosymbionts are capable of synthesizing tryptophan, which is required by the aphid host. PMID- 8407820 TI - Cloning and sequencing of a cellobiose phosphotransferase system operon from Bacillus stearothermophilus XL-65-6 and functional expression in Escherichia coli. AB - Cellulolytic strains of Bacillus stearothermophilus were isolated from nature and screened for the presence of activities associated with the degradation of plant cell walls. One isolate (strain XL-65-6) which exhibited strong activities with 4 methylumbelliferyl-beta-D-glucopyranoside (MUG) and 4-methylumbelliferyl-beta-D cellobiopyranoside (MUC) was used to construct a gene library in Escherichia coli. Clones degrading these model substrates were found to encode the cellobiose specific genes of the phosphoenolpyruvate-dependent phosphotransferase system (PTS). Both MUG and MUC activities were present together, and both activities were lost concurrently during subcloning experiments. A functional E. coli ptsI gene was required for MUC and MUG activities (presumably a ptsH gene also). The DNA fragment from B. stearothermophilus contained four open reading frames which appear to form a cel operon. Intergenic stop codons for celA, celB, and celC overlapped the ribosomal binding sites of the respective downstream genes. Frameshift mutations or deletions in celA, celB, and celD were individually shown to result in a loss of MUC and MUG activities. On the basis of amino acid sequence homology and hydropathy plots of translated sequences, celA and celB were identified as encoding PTS enzyme II and celD was identified as encoding PTS enzyme III. These translated sequences were remarkably similar to their respective E. coli homologs for cellobiose transport. No reported sequences exhibited a high level of homology with the celC gene product. The predicted carboxy-terminal region for celC was similar to the corresponding region of E. coli celF, a phospho-beta-glucosidase. An incomplete regulatory gene (celR) and proposed promoter sequence were located 5' to the proposed cel operon. A stem loop resembling a rho-independent terminator was present immediately downstream from celD. These results indicate that B. stearothermophilus XL-65-6 contains a cellobiose-specific PTS for cellobiose uptake. Similar systems may be present in other gram-positive bacteria. PMID- 8407821 TI - Genetic organization of a cluster of genes involved in the production of phaseolotoxin, a toxin produced by Pseudomonas syringae pv. phaseolicola. AB - Phaseolotoxin [N delta(N'-sulfo-diaminophosphinyl)-ornithyl-alanyl- homoarginine] produced by Pseudomonas syringae pv. phaseolicola, the bean halo blight pathogen, is a potent inhibitor of ornithine carbamoyltransferase (OCT). Inhibition of OCT in infected plants leads to chlorosis and growth inhibition. A genomic cosmid clone, pHK120, containing a 25-kb fragment of DNA from a wild-type strain of P. syringae pv. phaseolicola restores toxin production in Tox- mutants. Tn5 mutagenesis of pHK120 and marker exchange of pHK120::Tn5 plasmids in the wild type strain resulted in the isolation of 39 chromosomal mutants that harbor Tn5 insertions at known positions. Toxin bioassays revealed that 28 of the mutants, with Tn5 insertions distributed throughout the insert of pHK120, were Tox-, indicating that a functional locus for toxin production in each mutant was inactivated. Complementation analysis was done by testing for toxin production strains that carried a genomic Tn5 at one location and a plasmid-borne Tn5 at another location (pair complementation). Pair complementation analysis of nine marker exchange mutants and a random genomic Tn5 mutant revealed that there are a minimum of eight toxin loci (phtA through phtH) in pHK120. Mutants carrying Tn5 insertions in the phtA, phtD, and phtF loci were complemented by deletion subclones containing fragments from pHK120; mutants carrying Tn5 insertions in the phtC locus were partially complemented by a subclone, and mutants carrying Tn5 insertions in the phtB, phtE, phtG, and phtH loci were not complemented by any of the available subclones. A comparison of the insert from pHK120 with that from pRCP17, a clone reported previously (R. C. Peet, P. B. Lindgren, D. K. Wills, and N. J. Panopoulos, J. Bacteriol. 166:1096-1105, 1986) by another laboratory to contain some of the phaseolotoxin genes and the phaseolotoxin resistant OCT gene, revealed that the inserts in these two cosmids overlap but differ in important respects. PMID- 8407822 TI - A new lipopeptide biosurfactant produced by Arthrobacter sp. strain MIS38. AB - A biosurfactant termed arthrofactin produced by Arthrobacter species strain MIS38 was purified and chemically characterized as 3-hydroxydecanoyl-D-leucyl-D asparagyl-D-threonyl-D- leucyl-D-leucyl-D-seryl-L-leucyl-D-seryl-L-isoleucyl-L isoleucyl-L-as paragyl lactone. Surface activity of arthrofactin was examined, with surfactin as a control. Critical micelle concentration values of arthrofactin and surfactin were around 1.0 x 10(-5) M and 7.0 x 10(-5) M at 25 degrees C, respectively. Arthrofactin was found to be five to seven times more effective than surfactin. The minimum surface tension value of arthrofactin was 24 mN/m at a concentration higher than the critical micelle concentration. According to the oil displacement assay, arthrofactin was a better oil remover than synthetic surfactants, such as Triton X-100 and sodium dodecyl sulfate. Arthrofactin is one of the most effective lipopeptide biosurfactants. PMID- 8407823 TI - Dibenzofuran 4,4a-dioxygenase from Sphingomonas sp. strain RW1: angular dioxygenation by a three-component enzyme system. AB - Sphingomonas sp. strain RW1 synthesized a constitutive enzyme system that oxygenated dibenzofuran (DBF) to 2,2',3-trihydroxybiphenyl (THB). We purified this dibenzofuran 4,4a-dioxygenase system (DBFDOS) and found it to consist of four components which catalyzed three activities. Two isofunctional, monomeric flavoproteins (components A1 and A2; M(r) of about 44,000) transferred electrons from NADH to the second component (B; M(r) of about 12,000), a ferredoxin, which transported electrons to the heteromultimeric (alpha 2 beta 2) oxygenase component (C; M(r) of alpha, 45,000; M(r) of beta, 23,000). DBFDOS consumed 1 mol each of NADH, O2, and DBF, which was dioxygenated to about 1 mol of THB; no intermediate was observed. The reaction was thus the dioxygenation of DBF at the 4 and 4a positions to give a diene-diol-hemiacetal which rearomatized by spontaneous loss of a phenolate group to form THB. Components A1 and A2 each reduced dichlorophenolindophenol but had negligible activity with cytochrome c; each lost the yellow color, observed to be flavin adenine dinucleotide, upon purification. Component B, which transported electrons to the oxygenase or cytochrome c, had an N-terminal amino acid sequence with high homology to the putidaredoxin of cytochrome P-450cam. The oxygenase had the UV spectrum of a Rieske iron-sulfur center. We presume DBFDOS to be a class IIA dioxygenase system (EC 1.14.12.-), functionally similar to pyrazon dioxygenase. PMID- 8407824 TI - Genetic evidence for a functional relationship between Hsp104 and Hsp70. AB - The phenotypes of single Hsp104 and Hsp70 mutants of the budding yeast Saccharomyces cerevisiae provide no clue that these proteins are functionally related. Mutation of the HSP104 gene severely reduces the ability of cells to survive short exposures to extreme temperatures (thermotolerance) but has no effect on growth rates. On the other hand, mutations in the genes that encode Hsp70 proteins have significant effects on growth rates but do not reduce thermotolerance. The absence of a thermotolerance defect in S. cerevisiae Hsp70 mutants is puzzling, since the protein clearly plays an important role in thermotolerance in a variety of other organisms. In this report, examination of the phenotypes of combined Hsp104 and Hsp70 mutants uncovers similarities in the functions of Hsp104 and Hsp70 not previously apparent. In the absence of the Hsp104 protein, Hsp70 is very important for thermotolerance in S. cerevisiae, particularly at very early times after a temperature upshift. Similarly, Hsp104 plays a substantial role in vegetative growth under conditions of decreased Hsp70 protein levels. These results suggest a close functional relationship between Hsp104 and Hsp70. PMID- 8407825 TI - Characterization and transcriptional regulation of the 2'-N-acetyltransferase gene from Providencia stuartii. AB - We have cloned the chromosomally encoded 2'-N-acetyltransferase gene [aac(2')-Ia] from Providencia stuartii. DNA sequence analysis of the cloned insert identified a single open reading frame, which is capable of encoding a protein with a predicted molecular mass of 20,073 Da. The deduced AAC(2')-Ia protein showed no significant homology to other proteins, including all of the AAC(3) and AAC(6') proteins. Primer extension analysis was used to identify the aac(2')-Ia promoter, which contained an unusual sequence (CTTTTT) at the -35 region. Expression of the aac(2')-Ia gene occurs at low levels in wild-type P. stuartii strains; therefore, they are aminoglycoside susceptible. We have isolated mutants with high-level AAC(2')-Ia expression at a frequency of 4.8 x 10(-6). Detailed analysis of one mutant demonstrated a 12.2-fold increase in the accumulation of aac(2')-Ia mRNA. In addition, the levels of beta-galactosidase expression from a plasmid-encoded aac(2')-lacZ transcriptional fusion were increased 11.5-fold in this mutant relative to those in an isogenic wild-type strain. These results suggested that a trans-acting factor, designated aar (for aminoglycoside acetyltransferase regulator), controls AAC(2')-Ia expression in P. stuartii. PMID- 8407826 TI - Purification of the integration host factor homolog of Rhodobacter capsulatus: cloning and sequencing of the hip gene, which encodes the beta subunit. AB - We describe a method for rapid purification of the integration host factor (IHF) homolog of Rhodobacter capsulatus that has allowed us to obtain microgram quantities of highly purified protein. R. capsulatus IHF is an alpha beta heterodimer similar to IHF of Escherichia coli. We have cloned and sequenced the hip gene, which encodes the beta subunit. The deduced amino acid sequence (10.7 kDa) has 46% identity with the beta subunit of IHF from E. coli. In gel electrophoretic mobility shift DNA binding assays, R. capsulatus IHF was able to form a stable complex in a site-specific manner with a DNA fragment isolated from the promoter of the structural hupSL operon, which contains the IHF-binding site. The mutated IHF protein isolated from the Hup- mutant IR4, which is mutated in the himA gene (coding for the alpha subunit), gave a shifted band of greater mobility, and DNase I footprinting analysis has shown that the mutated IHF interacts with the DNA fragment from the hupSL promoter region differently from the way that the wild-type IHF does. PMID- 8407827 TI - Threonine formation via the coupled activity of 2-amino-3-ketobutyrate coenzyme A lyase and threonine dehydrogenase. AB - The enzymes L-threonine dehydrogenase and 2-amino-3-ketobutyrate coenzyme A (CoA) lyase are known to catalyze the net conversion of L-threonine plus NAD+ plus CoA to NADH plus glycine plus acetyl-CoA. When homogeneous preparations of these two enzymes from Escherichia coli were incubated together for 40 min at 25 degrees C with glycine, acetyl-CoA, and NADH, a 36% decrease in the level of glycine (with concomitant NADH oxidation) was matched by formation of an equivalent amount of threonine, indicating that this coupled sequence of enzyme-catalyzed reactions is reversible in vitro. Several experimental factors that affect the efficiency of this conversion in vitro were examined. A constructed strain of E. coli, MD901 (glyA thrB/C tdh), was unable to grow unless both glycine and threonine were added to defined rich medium. Introduction of the plasmid pDR121 (tdh+kbl+) into this strain enabled the cells to grow in the presence of either added glycine or threonine, indicating that interconversion of these two amino acids occurred. Threonine that was isolated from the total pool of cellular protein of MD901/pDR121 had the same specific radioactivity as the [14C]glycine added to the medium, establishing that threonine was formed exclusively from glycine in this strain. Comparative growth rate studies with several strains of E. coli containing plasmid pDR121, together with the finding that kcat values of pure E. coli 2-amino-3-ketobutyrate CoA lyase favor the cleavage of 2-amino-3 ketobutyrate over its formation by a factor of 50, indicate that the biosynthesis of threonine is less efficient than glycine formation via the coupled threonine dehydrogenase-2-amino-3-ketobutyrate lyase reactions. PMID- 8407828 TI - recA mutations that reduce the constitutive coprotease activity of the RecA1202(Prtc) protein: possible involvement of interfilament association in proteolytic and recombination activities. AB - Twenty-eight recA mutants, isolated after spontaneous mutagenesis generated by the combined action of RecA1202(Prtc) and UmuDC proteins, were characterized and sequenced. The mutations are intragenic suppressors of the recA1202 allele and were detected by the reduced coprotease activity of the gene product. Twenty distinct mutation sites were found, among which two mutations, recA1620 (V-275- >D) and recA1631 (I-284-->N), were mapped in the C-terminal portion of the interfilament contact region (IFCR) in the RecA crystal. An interaction of this region with the part of the IFCR in which the recA1202 mutation (Q-184-->K) is mapped could occur only intermolecularly. Thus, altered IFCR and the likely resulting change in interfilament association appear to be important aspects of the formation of a constitutively active RecA coprotease. This observation is consistent with the filament-bundle theory (R. M. Story, I. T. Weber, and T. A. Steitz, Nature (London) 335:318-325, 1992). Furthermore, we found that among the 20 suppressor mutations, 3 missense mutations that lead to recombination defective (Rec-) phenotypes also mapped in the IFCR, suggesting that the IFCR, with its putative function in interfilament association, is required for the recombinase activity of RecA. We propose that RecA-DNA complexes may form bundles analogous to the RecA bundles (lacking DNA) described by Story et al. and that these RecA-DNA bundles play a role in homologous recombination. PMID- 8407829 TI - Mobilization of small plasmids in Bacillus thuringiensis subsp. israelensis is accompanied by specific aggregation. AB - Mobilizations of pBC16 and pAND006, containing the replicon of the Bacillus thuringiensis subsp. israelensis plasmid pTX14-3, between strains of B. thuringiensis subsp. israelensis were examined. Transconjugants appeared after a few minutes and reached a maximum frequency after approximately 2 h. Plasmid pBC16 was mobilized at a frequency approximately 200 times that of pAND006. However, pAND006 was consistently transferred, suggesting that the replicon of pTX14-3 is sufficient to sustain mobilization in B. thuringiensis subsp. israelensis. A specific protease-sensitive coaggregation between strains of B. thuringiensis subsp. israelensis was found to be unambiguously correlated with plasmid transfer. Two aggregation phenotypes, Agr+ and Agr-, were identified in this subspecies. Aggregation disappeared when the optical density of the mating mixture at 600 nm exceeded approximately 1, and it did not reappear upon dilution. Aggregation was shown to involve interactions of cells with opposite aggregation phenotypes, and evidence of a proteinaceous molecule on the surface of the Agr- that is cells involved in aggregation formation is presented. Matings and selection for the presence of two antibiotic resistance plasmids followed by identification of the host cell revealed that mobilization was unidirectional, from the Agr+ cell to the Agr- cell. The aggregation phenotype was found to be transferred with high frequency (approximately 100%) in broth matings, and the appearance of Agr- isolates from Agr+ strains suggested that the loci involved in aggregation formation are located on a plasmid. No excreted aggregation-inducing signals were detected in the supernatant or culture filtrate of either the donor, the recipient, or the mating mixture. PMID- 8407830 TI - Three distinct chromosome replication states are induced by increasing concentrations of DnaA protein in Escherichia coli. AB - The DnaA protein concentration in Escherichia coli was increased above the wild type level by inducing a lacP-controlled dnaA gene located on a plasmid. In these cells with different DnaA protein levels, we measured several parameters: dnaA gene expression; cell size, amount of DNA per cell, and number of origins per cell by flow cytometry; and origin-to-terminus ratio and the frequencies of five other markers on the chromosome by Southern hybridization. The response of the cells to higher levels of DnaA protein could be divided into three states. From the normal level to a level 1.5-fold higher, DnaA protein had little effect on dnaA gene expression and the rate of DNA replication but led to nearly proportional increases in DNA and origin concentrations. Between 1.5- and 3-fold, the normal DnaA protein concentration, dnaA gene expression was gradually decreased. In this interval, the origin concentration increased significantly; however, the replication rate was severely affected, becoming slower--especially near the origin--the higher the DnaA protein concentration, and as a result, the DNA concentration was constant. Further increases in the DnaA protein concentration did not lead to an increased origin concentration. Thus, the initiation mass was set by the DnaA protein from the normal level to an at least twofold-increased level, but the increased initiation did not lead to a large increase in the amount of DNA per unit of mass because of the inhibition of replication fork velocity. PMID- 8407831 TI - Functional exchangeability of the ABC proteins of the periplasmic binding protein dependent transport systems Ugp and Mal of Escherichia coli. AB - The periplasmic binding protein-dependent transport systems Ugp and Mal of Escherichia coli transport sn-glycerol-3-phosphate and maltose, respectively. The UgpC and MalK proteins of these transport systems, which couple energy to the transport process by ATP-hydrolysis, are highly homologous, suggesting that they might be functionally exchangeable. Complementation experiments showed that UgpC expression could restore growth of a malK mutant on maltose as a carbon source, provided that it was expressed at a sufficiently high level in the absence of the integral inner membrane components UgpA and/or UgpE of the Ugp system. Conversely, MalK expression could complement ugpC mutants and restore the utilization of sn-glycerol-3-phosphate as a phosphate source. The hybrid transporters appeared to be less efficient than the wild-type systems. The complementation of ugpC mutations by MalK was strongly inhibited by the presence of glucose or alpha-methylglucoside, which are substrates of the phosphotransferase system. This inhibition is probably due to hypersensitivity of the hybrid UgpBAE-MalK transporter to inducer exclusion. UgpC expression did not complement the regulatory function of MalK in mal gene expression. The exchangeability of UgpC and MalK indicates that these proteins do not contribute to a substrate-binding site conferring substrate specificity to the transporter. These are the first examples of functional, hybrid periplasmic permeases in which the energy-coupling components could be functionally exchanged. PMID- 8407832 TI - Isolation and characterization of a DNA replication origin from the 1,700 kilobase-pair symbiotic megaplasmid pSym-b of Rhizobium meliloti. AB - A 4-kb fragment active as an autonomously replicating sequence (ARS) from the Rhizobium meliloti symbiotic megaplasmid pSym-b was isolated by selecting for sequences that allowed a normally nonreplicative pBR322 derivative to replicate in R. meliloti. The resulting Escherichia coli-R. meliloti shuttle plasmid (mini pSym-b) containing the ARS also replicated in the closely related Agrobacterium tumefaciens, but only in strains carrying pSym-b, suggesting that a megaplasmid encoded trans-acting factor is required. The copy number of mini-pSym-b was approximately the same as that of the resident megaplasmid, and mini-pSym-b was unstable in the absence of antibiotic selection. An 0.8-kb DNA subfragment was sufficient for replication in both R. meliloti and A. tumefaciens. The minimal ARS exhibited several sequence motifs common to other replication origins, such as an AT-rich region, three potential DnA binding sites, a potential 13-mer sequence, and several groups of short direct repeats. Hybridization experiments indicated that there may be a related ARS on the other megaplasmid, pSym-a. The pSym-b ARS was mapped near exoA, within a region nonessential for pSym-b replication. These results suggest that the R. meliloti megaplasmids share conserved replication origins and that pSym-b contains multiple replication origins. Since the mini-pSym-b shuttle vector can coexist with IncP-1 broad-host range plasmids, it is also now possible to use two compatible plasmids for cloning and genetic manipulation in R. meliloti. PMID- 8407833 TI - A cdc-like autolytic Saccharomyces cerevisiae mutant altered in budding site selection is complemented by SPO12, a sporulation gene. AB - LYT1 is an essential gene for the growth and morphogenesis of Saccharomyces cerevisiae. A detailed characterization of mutants carrying the lyt1-1 allele showed that this mutation was recessive and pleiotropic, affecting both mitotic and meiotic functions. At the nonpermissive temperature of 37 degrees C, lyt1 haploid strains budded at a distal position (instead of an axial one, as in wild type haploid strains) and underwent autolysis when the buds were almost the size of the mother cells. These mitotic alterations in cell stability and budding topology were dependent on growth and protein synthesis. Autolysis was prevented by inhibiting DNA synthesis (with hydroxyurea) or by blocking the assembly of microtubules (with benomyl), suggesting that loss of cell viability must occur at a fixed mitotic cycle stage after DNA synthesis and mitotic spindle assembly. On the other hand, lyt1-1/lyt1-1 diploids failed to sporulate at both 24 and 37 degrees C. Taking into account these characteristics, the lyt1 mutant could be considered a cdc-like mutant. By genetic transformation of an appropriate lyt1 strain with a genomic library, ligated to the multicopy vector YEp13, we isolated a gene capable of complementing mitotic alterations but not the meiotic defect. This was the sporulation-specific gene SPO12, which is expressed under the control of the locus MAT in meiosis and is also expressed in the mitotic cycle (V. Parkes and L. H. Johnston, Nucleic Acids Res. 20:5617-5623, 1992). A significant level of SPO12 mRNA can be detected when this gene is inserted in a multicopy plasmid. PMID- 8407834 TI - An Escherichia coli gene in search of a function: phenotypic effects of the gene recently identified as murI. AB - Earlier we reported that an open reading frame located at 89.5 min of the Escherichia coli map (ORFI) codes for a protein of unknown function that could be overexpressed and purified to homogeneity (G. Baliko, A. Raukas, I. Boros, and P. Venetianer, Mol. Gen. Genet. 211:326-331, 1988). In the work described here, we attempted to learn the function of this protein by inactivating the chromosomal gene and providing it or its deletion derivatives on temperature-sensitive plasmids. We found that the presence of the functional ORFI gene is essential; cells are not viable at the nonpermissive temperature or when the region coding for the C-terminal 50 amino acids of the protein is deleted. At intermediate temperatures or when the gene is overexpressed, characteristic changes occur in cell morphology, nucleoid separation during cell division, and supercoiling of plasmids. The possible mechanisms of these effects are discussed in view of the fact that Doublet et al. (P. Doublet, J. van Heijenoort, and D. Mengin-Lecreulx, J. Bacteriol. 174:5772-5779, 1992) recently identified the ORFI gene as murI, involved in D-glutamic acid biosynthesis. PMID- 8407835 TI - Excision, transfer, and integration of NBU1, a mobilizable site-selective insertion element. AB - The Bacteroides species harbor a family of conjugative transposons called tetracycline resistance elements (Tcr elements) that transfer themselves from the chromosome of a donor to the chromosome of a recipient, mobilize coresident plasmids, and also mediate the excision and circularization of members of a family of 10- to 12-kbp insertion elements which share a small region of DNA homology and are called NBUs (for nonreplicating Bacteroides units). The NBUs are sometimes cotransferred with Tcr elements, and it was postulated previously that the excised circular forms of the NBUs were plasmidlike forms and were transferred like plasmids and then integrated into the recipient chromosome. We used chimeric plasmids containing one of the NBUs, NBU1, and a Bacteroides Escherichia coli shuttle vector to show that this hypothesis is probably correct. NBU1 contained a region that allowed mobilization by both the Tcr elements and IncP plasmids, and we used these conjugal elements to allow us to estimate the frequencies of excision, mobilization, and integration of NBU1 in Bacteroides hosts to be approximately 10(-2), 10(-5) to 10(-4), and 10(-2), respectively. Although functions on the Tcr elements were required for the excision circularization and mobilization of NBU1, no Tcr element functions were required for integration into the recipient chromosome. Analysis of the DNA sequences at the integration region of the circular form of NBU1, the primary insertion site in the Bacteroides thetaiotaomicron 5482 chromosome, and the resultant NBU1 chromosome junctions showed that NBU1 appeared to integrate into the primary insertion site by recombining within an identical 14-bp sequence present on both NBU1 and the target, thus leaving a copy of the 14-bp sequence at both junctions. The apparent integration mechanism and the target selection of NBU1 were different from those of both XBU4422, the only member of the conjugal Tcr elements for which these sequences are known, and Tn4399, a mobilizable Bacteroides transposon. The NBUs appear to be a distinct type of mobilizable insertion element. PMID- 8407836 TI - Characterization of the mobilization region of a Bacteroides insertion element (NBU1) that is excised and transferred by Bacteroides conjugative transposons. AB - Many Bacteroides clinical isolates carry large conjugative transposons that, in addition to transferring themselves, excise, circularize, and transfer smaller, unlinked chromosomal DNA segments called NBUs (nonreplicating Bacteroides units). We report the localization and DNA sequence of a region of one of the NBUs, NBU1, that was necessary and sufficient for mobilization by Bacteroides conjugative transposons and by IncP plasmids. The fact that the mobilization region was internal to NBU1 indicates that the circular form of NBU1 is the form that is mobilized. The NBU1 mobilization region contained a single large (1.4-kbp) open reading frame (ORF1), which was designated mob. The oriT was located within a 220 bp region upstream of mob. The deduced amino acid sequence of the mob product had no significant similarity to those of mobilization proteins of well-characterized Escherichia coli group plasmids such as RK2 or of either of the two mobilization proteins of Bacteroides plasmid pBFTM10. There was, however, a high level of similarity between the deduced amino acid sequence of the mob product and that of the product of a Bacteroides vulgatus cryptic open reading frame closely linked to a cefoxitin resistance gene (cfxA). PMID- 8407837 TI - Presence of protein constituents of the gram-positive bacterial phosphotransferase regulatory system in Acholeplasma laidlawii. AB - Acholeplasma species have been reported to lack a functional phosphoenolpyruvate:sugar phosphotransferase system (PTS). We show here that Acholeplasma laidlawii possesses activities of enzyme I, HPr, HPr(ser) kinase, and HPr(ser-P) phosphatase but lacks detectable activities of enzymes II of the PTS. HPr from this organism was purified, and the regulatory properties of the kinase and phosphatase were characterized and shown to differ from those of previously studied bacteria. The results suggest the presence of an incomplete PTS in A. laidlawii which has the potential to function in a unique regulatory capacity. PMID- 8407838 TI - An efficient approach to identify ilvA mutations reveals an amino-terminal catalytic domain in biosynthetic threonine deaminase from Escherichia coli. AB - High-level expression of the regulatory enzyme threonine deaminase in Escherichia coli strains grown on minimal medium that are deficient in the activities of enzymes needed for branched-chain amino acid biosynthesis result in growth inhibition, possibly because of the accumulation of toxic levels of alpha ketobutyrate, the product of the committed step in isoleucine biosynthesis. This condition affords a means for selecting genetic variants of threonine deaminase that are deficient in catalysis by suppression of growth inhibition. Strains harboring mutations in ilvA that decreased the catalytic activity of threonine deaminase were found to grow more rapidly than isogenic strains containing wild type ilvA. Modification of the ilvA gene to introduce additional unique, evenly spaced restriction enzyme sites facilitated the identification of suppressor mutations by enabling small DNA fragments to be subcloned for sequencing. The 10 mutations identified in ilvA code for enzymes with significantly reduced activity relative to that of wild-type threonine deaminase. Values for their specific activities range from 40% of that displayed by wild-type enzyme to complete inactivation as evidenced by failure to complement an ilvA deletion strain to isoleucine prototrophy. Moreover, some mutant enzymes showed altered allosteric properties with respect to valine activation and isoleucine inhibition. The location of the 10 mutations in the 5' two-thirds of the ilvA gene is consistent with suggestions that threonine deaminase is organized functionally with an amino terminal domain that is involved in catalysis and a carboxy-terminal domain that is important for regulation. PMID- 8407839 TI - A chromosomally encoded two-component sensory transduction system is required for virulence of Agrobacterium tumefaciens. AB - TnphoA mutagenesis of Agrobacterium tumefaciens identified new extracytoplasmic protein-encoding virulence loci. Mutations in these loci conferred increased sensitivity to detergents and several antibiotics. Clones carrying these loci were isolated from an A. tumefaciens cosmid library by complementation of the detergent sensitivities of the mutants. The locus on one complementing clone was delineated by Tn5 and TnphoA mutagenesis. DNA sequence analysis of the delineated region revealed that this locus is made up of two transcriptional units, chvG and chvI, which were predicted, on the basis of amino acid sequence homology, to encode the members of a two-component sensory transduction system. The membrane spanning sensor, a histidine protein kinase, was designated ChvG, and the response regulator, presumably a transcriptional activator, was designated ChvI. Surprisingly, ChvG was also predicted to contain a Walker type A consensus nucleotide binding site, which is unusual for sensor histidine protein kinases. Site-specific insertion mutations in either chvG or chvI abolished tumor formation ability, as well as the ability to grow on complex media. Neither the genes which are regulated nor the inducing signal is known yet for this system. PMID- 8407840 TI - The chromosomal response regulatory gene chvI of Agrobacterium tumefaciens complements an Escherichia coli phoB mutation and is required for virulence. AB - In an effort to identify the Agrobacterium tumefaciens phosphate regulatory gene(s), we isolated a clone from an A. tumefaciens cosmid library that restored regulated alkaline phosphatase activity to an Escherichia coli phoB mutant. The gene that complemented phoB was localized by subcloning and deletion analysis, and the DNA sequence was determined. An open reading frame, denoted chvI, was identified that encoded a predicted protein with amino acid similarity to the family of bacterial response regulators and 35% identify to PhoB. Surprisingly, an A. tumefaciens chvI mutant showed normal induction of phosphatase activity and normal virG expression when grown in phosphate-limiting media. However, this mutant was unable to grow in media containing tryptone, peptone, or Casamino Acids and was also more sensitive than the wild type to acidic extracellular pH. This mutant was avirulent on Kalanchoee diagremontiana and was severely attenuated in vir gene expression. The pH-inducible expression of virG was also abolished. Growth of the chvI mutant was inhibited by K. diagremontiana wound sap, suggesting that avirulence may be due, in part, to the inability of this mutant to survive the plant wound environment. PMID- 8407841 TI - Construction of an SfiI macrorestriction map of the Candida albicans genome. AB - The opportunistic fungal pathogen, Candida albicans, is diploid as usually isolated and has no apparent sexual cycle. Genetic analysis has therefore been very difficult. Molecular genetics has yielded important information in the past few years, but it too is hampered by the lack of a good genetic map. Using the well-characterized strain 1006 and strain WO-1, which undergoes the white-opaque phenotypic transition, we have developed a genomic restriction map of C. albicans with the enzyme SfiI. There are approximately 34 SfiI restriction sites in the C. albicans genome. Restriction fragments were separated by pulsed-field electrophoresis and were assigned to chromosomes by hybridization of complete and partial digests with known chromosome-specific probes as well as by digestion of isolated chromosomes. Telomeric fragments were identified by hybridization with a telomere-specific probe (C. Sadhu, M.J. McEachern, E.P. Rustchenko-Bulgac, J. Schmid, D.R. Soll, and J.B. Hicks, J. Bacteriol. 173:842-850, 1991). WO-1 differs from 1006 in that it has undergone three reciprocal chromosomal translocations. Analysis of the translocation products indicates that each translocation has occurred at or near an SfiI site; thus, the SfiI fragments from the two strains are similar or identical. The tendency for translocation to occur at or near SfiI sites may be related to the repeated sequence RPS 1, which contains four such sites and could provide homology for ectopic pairing and crossing over. The genome size of both strains is about 16 to 17 megabases, in good agreement with previous determinations. PMID- 8407842 TI - Fusion of Spiroplasma floricola cells with small unilamellar vesicles is dependent on the age of the culture. AB - Small unilamellar vesicles were labeled with the fluorescent probe octadecylrhodamine B chloride and mixed with intact Spiroplasma floricola cells. The increase in fluorescence observed was interpreted as a result of the dilution of the probe in the unlabeled S. floricola membranes because of lipid mixing upon fusion. The progression of S. floricola cultures to the stationary phase of growth was accompanied by a sharp decrease in the ability of the cells to fuse with small unilamellar vesicles. Low fusogenic activity was also detected in cells from cultures that were aged in a growth medium maintained at pH 7.5 throughout the growth cycle. Chemical analysis of the cell membrane preparations isolated from cells harvested at the various phases of growth revealed that the phospholipid content and composition and the cholesterol/phospholipid molar ratio were changed very little upon aging of the cultures. Likewise, no changes in the fatty acid composition of membrane lipids were detected, with palmitic and oleic acids predominating throughout the cycle. Nonetheless, upon aging of S. floricola cultures, a pronounced increase in the levels of both cholesteryl esters, incorporated from the growth medium, and organic peroxides was observed. A decrease in both fluorescence anisotropy of diphenylhexatriene and merocyanine 540 binding to membranes of aged cells was also detected. The possible influence of these changes on the fusogenic activity of the cells is discussed. PMID- 8407843 TI - Three overlapping lct genes involved in L-lactate utilization by Escherichia coli. AB - In Escherichia coli, the lct locus at min 80 on the chromosome map is associated with ability to grow on L-lactate and to synthesize a substrate-inducible flavin linked dehydrogenase. Similar to that of the glpD-encoded aerobic glycerol-3 phosphate dehydrogenase, the level of induced enzyme activity is elevated by aerobiosis. Both of these controls are mediated by the two-component signal transduction system ArcB/ArcA, although sensitivity to the control is much more striking for L-lactate dehydrogenase. This study disclosed that the lct locus contained three overlapping genes in the clockwise order of lctD (encoding a flavin mononucleotide-dependent dehydrogenase), lctR (encoding a putative regulator), and lctP (encoding a permease) on the chromosomal map. These genes, however, are transcribed in the counterclockwise direction. No homology in amino acid sequence was found between aerobic glycerol-3-phosphate dehydrogenase and L lactate dehydrogenase. A phi (lctD-lac) mutant was inducible by L-lactate but not D-lactate. Although the mutant lost the ability to grow on L-lactate, growth on D lactate, known to depend on a different enzyme, remained normal. PMID- 8407844 TI - A phase variant of Bordetella pertussis with a mutation in a new locus involved in the regulation of pertussis toxin and adenylate cyclase toxin expression. AB - A novel nonhemolytic phase variant of Bordetella pertussis was characterized. This strain is strongly impaired in the transcription of the pertussis and adenylate cyclase toxins, whereas other known virulence-related factors such as the filamentous hemagglutinin, the fimbriae, and the outer membrane protein pertactin are expressed and regulated normally. Complementation and allelic exchange experiments demonstrated that the mutation is localized neither in the bvg locus involved in virulence regulation nor in the genes responsible for synthesis and transport of the toxins pertussis and adenylate cyclase. Instead, the mutation impairing transcription of at least the two toxin genes is located in a new genetic locus, which acts together with the BvgA/S two-component regulatory system on the expression of a subset of virulence genes. Further analysis suggested that most presumably the mutation affects a sequence-specific DNA-binding protein which contributes to transcriptional activation. The mutant was nonlethal in a murine respiratory model, which corresponds well with the lack of expression of the toxins. However, the clearing rate of this mutant from the lungs of mice was much lower than that of a bvg mutant, suggesting that factors other than the toxins may play a role in the persistence of the bacteria in the respiratory tract of mice. PMID- 8407845 TI - Isolation and analysis of eight exe genes and their involvement in extracellular protein secretion and outer membrane assembly in Aeromonas hydrophila. AB - The exeE gene of Aeromonas hydrophila has been shown to be required for the secretion of most if not all of the extracellular proteins produced by this bacterium. In addition, an exeE::Tn5-751 insertion mutant of A. hydrophila was found to be deficient in the amounts of a number of its major outer membrane proteins (B. Jiang and S. P. Howard, J. Bacteriol. 173:1241-1249, 1991). The exeE gene and the exeF gene were previously isolated as part of a fragment which complemented this mutant. In this study, we have isolated and sequenced a further eight exe genes, exeG through exeN, which constitute the 3' end of the exe operon. These genes have a high degree of similarity with the extracellular secretion operons of a number of different gram-negative bacteria. Marker exchange mutagenesis was used to insert kanamycin resistance cassettes into three different regions of the exe operon. The phenotypes of these mutants showed that in A. hydrophila this operon is required not only for extracellular protein secretion but also for normal assembly of the outer membrane, in particular with respect to the quantities of the major porins. Five of the Exe proteins contain type IV prepilin signal sequences, although the prepilin peptidase gene does not appear to form part of the exe operon. Limited processing of the ExeG protein was observed when it was expressed in Escherichia coli, and this processing was greatly accelerated in the presence of the prepilin peptidase of Pseudomonas aeruginosa. PMID- 8407846 TI - Penicillin-binding protein 2 inactivation in Escherichia coli results in cell division inhibition, which is relieved by FtsZ overexpression. AB - Aminoacyl-tRNA synthetase mutants of Escherichia coli are resistant to amdinocillin (mecillinam), a beta-lactam antibiotic which specifically binds penicillin-binding protein 2 (PBP2) and prevents cell wall elongation with concomitant cell death. The leuS(Ts) strain, in which leucyl-tRNA synthetase is temperature sensitive, was resistant to amdinocillin at 37 degrees C because of an increased guanosine 5'-diphosphate 3'-diphosphate (ppGpp) pool resulting from partial induction of the stringent response, but it was sensitive to amdinocillin at 25 degrees C. We constructed a leuS(Ts) delta (rodA-pbpA)::Kmr strain, in which the PBP2 structural gene is deleted. This strain grew as spherical cells at 37 degrees C but was not viable at 25 degrees C. After a shift from 37 to 25 degrees C, the ppGpp pool decreased and cell division was inhibited; the cells slowly carried out a single division, increased considerably in volume, and gradually lost viability. The cell division inhibition was reversible when the ppGpp pool increased at high temperature, but reversion required de novo protein synthesis, possibly of septation proteins. The multicopy plasmid pZAQ, overproducing the septation proteins FtsZ, FtsA, and FtsQ, conferred amdinocillin resistance on a wild-type strain and suppressed the cell division inhibition in the leuS(Ts) delta (rodA-pbpA)::Kmr strain at 25 degrees C. The plasmid pAQ, in which the ftsZ gene is inactivated, did not confer amdinocillin resistance. These results lead us to hypothesize that the nucleotide ppGpp activates ftsZ expression and thus couples cell division to protein synthesis. PMID- 8407847 TI - Isolation of a constitutively expressed enzyme for hydrolysis of carbaryl in Pseudomonas aeruginosa. AB - A hydrolase constitutively expressed in Pseudomonas aeruginosa which converts carbaryl to 1-naphthol was purified 1,767-fold by using a combination of anion exchange, hydroxylapatite, gel filtration, and hydrophobic interaction chromatography techniques. The presence of Triton X-100 in buffers was necessary for deaggregation and purification of the hydrolase. This is the first membrane bound hydrolase involved in the hydrolysis of any methylcarbamate pesticide purified from a bacterial source to date. The enzyme exhibited a unique specificity of hydrolyzing only carbaryl (1-naphthyl N-methylcarbamate) but not any other methylcarbamates. The purified enzyme was a monomer with a molecular mass of 65,000 Da. The pH and temperature optima for the enzyme activity were 8.5 and 45 degrees C, respectively. No cofactor requirement for the hydrolase activity could be demonstrated, and none of the divalent cations studied affected the activity of the enzyme. Also, the enzyme activity was not affected by the thiols: dithioerythritol, dithiothreitol, and 2-mercaptoethanol. The Km and Vmax values for carbaryl were 9 microM and 7.9 mumol/min/mg of protein, respectively. PMID- 8407848 TI - Identification of an intracellular pyrimidine-specific endoribonuclease from Bacillus subtilis. AB - Two intracellular RNases which were easily separated by fractionation on strong anion- or cation-exchange resins were identified from Bacillus subtilis. One cleaved any phosphodiester bond, while the second cleaved only pyrimidine-N bonds. The enzyme with pyrimidine-N specificity was approximately 15 kDa, had a pH optimum of approximately 6.2, degraded C-C bonds approximately 10 times faster than U-U bonds, and was completely inactive against single-stranded DNA. The enzyme is called RNase C and may be the first reported broad-specificity endoribonuclease from B. subtilis. PMID- 8407849 TI - Effects of ribosome-inactivating proteins on Escherichia coli and Agrobacterium tumefaciens translation systems. AB - The effects of 30 type 1 and of 2 (ricin and volkensin) type 2 ribosome inactivating proteins (RIPs) on Escherichia coli and Agrobacterium tumefaciens cell-free translation systems were compared with the effects on a rabbit reticulocyte translation system. The depurinating activity of RIPs on E. coli ribosomes was also evaluated. Only six type 1 RIPs inhibited endogenous mRNA directed translational activity of E. coli lysates, with submicromolar 50% inhibitory concentrations. Four RIPs had similar activities on poly(U)-directed phenylalanine polymerization by E. coli ribosomes, and three RIPs inhibited poly(U)-directed polyphenylalanine synthesis by A. tumefaciens ribosomes, with submicromolar 50% inhibitory concentrations. PMID- 8407850 TI - Coexpression of colanic acid and serotype-specific capsular polysaccharides in Escherichia coli strains with group II K antigens. AB - In Escherichia coli K-12, the rcsA and rcsB gene products are positive regulators in expression of the slime polysaccharide colanic acid. We have previously demonstrated the presence of rcsA sequences in E. coli K1 and K5, strains with group II capsular K antigens, and shown that introduction of multicopy rcsA into these strains results in the expression of colanic acid. We report here the presence of rcsB sequences in E. coli K1 and K5 and demonstrate that RcsB also plays a role in the biosynthesis of colanic acid in strains with group II K antigens. In E. coli K1 and K5 grown at 37 degrees C, multicopy rcsB and the resulting induction of colanic acid synthesis had no significant effect on synthesis of the group II K antigens. K-antigen-specific sugar transferase activities were not significantly different in the presence or absence of multicopy rcsB, and introduction of a cps mutation to eliminate colanic acid biosynthesis in a K1-derivative strain did not influence the activity of the polysialyltransferase enzyme responsible for synthesis of the K1 polymer. Furthermore, immunoelectron microscopy showed no detectable difference in the size or distribution of the group II K-antigen capsular layer in cells which produced colanic acid. Colanic acid expression therefore does not appear to significantly affect synthesis of the group II K-antigen capsule and, unlike for group I K antigens, expression of group II K antigens is not positively regulated by the rcs system. PMID- 8407851 TI - Absence of a direct role for RNase HI in initiation of DNA replication at the oriC site on the Escherichia coli chromosome. AB - On the basis of the experiments carried out with rnhA224 mutants, we previously concluded that RNase HI is not essential for initiation of Escherichia coli chromosome replication at oriC (T. Kogoma, N.L. Subia, and K. von Meyenburg, Mol. Gen. Genet. 200:103-109, 1985). In light of the recent finding that rnhA224 is a UGA nonsense mutation which can be leaky in certain genetic backgrounds, we reexamined this conclusion with the use of rnhA339 (Null)::cat mutants. The possibility that recB+ is required for initiation at the alternative origins (oriKs) of replication in rnhA mutants was also tested. The results clearly indicated that RNase HI is not essential for oriC initiation and that recB+ is not required for initiation at oriK sites. PMID- 8407852 TI - Tourette's syndrome and attention-deficit hyperactivity disorder: evidence for a genetic relationship. AB - BACKGROUND: Attention deficit disorder with or without hyperactivity (ADD) is present in 49% to 83% of patients with Tourette's syndrome (TS), and up to 50% of children with attention-deficit hyperactivity disorder (ADHD) have chronic tics or a family history of chronic tics. Two explanations have been offered for this association: (1) ADHD is part of the pleiotropic expression of the Gilles de la Tourette (Gts) gene(s), or (2) the presence of ADHD in TS is due to ascertainment bias. METHOD: To avoid ascertainment bias, we examined 338 first-degree relatives of 131 TS probands utilizing a structured questionnaire and the DSM-III criteria for ADD and DSM-III-R criteria for ADHD. All probands and many relatives were personally interviewed. RESULTS: Of the relatives with TS, 61% had ADD and 36% had ADHD. Of the relatives with chronic tics, 41% had ADD and 26% had ADHD. Log linear analysis showed a major, significant association between tics and ADHD. CONCLUSION: These results indicate that ADHD is part of the pleiotropic expression of the Gts gene(s). PMID- 8407853 TI - Dexamethasone suppression test and onset of poststroke depression in patients with ischemic infarction. AB - BACKGROUND: Studies documenting abnormal results in the 1.0-mg dexamethasone suppression test (DST) and the prevalence of depression in stroke patients are usually accomplished between 1 and 6 months poststroke. One study, however, reported that 27% of patients met DSM-III criteria for major depression and 20% for minor depression 1 to 3 weeks poststroke even though previous research has indicated that the prevalence of poststroke depression was greatest at approximately 6 months. Therefore, we decided to assess DST abnormalities and depression within the first month of stroke. METHOD: Twelve patients with single, computed tomography (CT)-verified, ischemic infarctions were administered the DST at 1 and 3 weeks poststroke. Each patient also received a complete psychiatric evaluation, including a special clinical interview and the 17-item Hamilton Rating Scale for Depression (HAM-D). 3 to 4 weeks poststroke. RESULTS: DST results were abnormal in 75% of the patients at 1 week poststroke and 50% of the patients at 3 weeks poststroke. Those patients whose HAM-D scores revealed more depressive symptoms at 3 to 4 weeks were more likely to evidence abnormal DST results (cortisol nonsuppression). None of the patients, however, met either DSM III or modified criteria for clinical depression at 3 to 4 weeks. CONCLUSION: Poststroke depression appears to have a delayed clinical onset. Abnormal DST results at 3 weeks poststroke may serve as a potential marker for those patients at risk for developing poststroke depression. PMID- 8407854 TI - d-Amphetamine versus methylphenidate effects in depressed inpatients. AB - BACKGROUND: Both d-amphetamine and methylphenidate cause similar behavioral and subjective effects and can relieve depressive symptoms in some patients. However, the neurochemical effects of d-amphetamine and methylphenidate are distinct, and clinical experience suggests that individual depressed patients may respond dissimilarly to each stimulant. METHOD: d-Amphetamine and methylphenidate were administered sequentially to 18 DSM-III diagnosed depressed inpatients on consecutive days, and the acute effects were measured by a variety of instruments. RESULTS: Although many patients were acutely better after taking one drug or the other, as assessed by Hamilton Rating Scale for Depression and Global Visual Analogue Scale scores, relatively few responded with equal improvement to both. CONCLUSION: Acute symptomatic improvement after d-amphetamine or methylphenidate is unpredictable and can only be determined by an empirical trial on an individual basis. These data also suggest that even if one stimulant is not helpful therapeutically, another might be. The long-term benefits of selective stimulant therapy in depressed patients remain to be assessed. PMID- 8407855 TI - Contribution of anticholinergic medication to symptom change in schizophrenia. PMID- 8407856 TI - Pharmacokinetics of antidepressants: why and how they are relevant to treatment. AB - The pharmacokinetics of antidepressants are reviewed with special emphasis on clinical applicability. There are substantial pharmacokinetic differences among the five major classes. Pharmacokinetic differences between members of a specific class are usually modest except for serotonin selective reuptake inhibitors. Between members of this class pharmacokinetic differences are the major distinguishing variables and have clinically important consequences. PMID- 8407857 TI - Introduction. Pharmacokinetics of psychotropic agents: why and how they are relevant to treatment. PMID- 8407858 TI - Pharmacokinetics of psychotropic drugs in special populations. AB - Patients treated for psychiatric illness may have a variety of characteristics that alter the pharmacokinetic profiles of psychotropic drugs. Genetic background, age, health status, and personal habits can change the body's ability to absorb, distribute, and metabolize medications. Most psychotropic drugs, with the exception of lithium, are eliminated via biotransformation in the liver rather than renal excretion, and a number of studies have demonstrated distinct phenotypes for hepatic metabolism involving the cytochrome P450 system. "Slow" metabolizers are likely to develop higher plasma concentrations of several different classes of psychotropic drugs, including tricyclic antidepressants. Advancing age reduces renal function but has little effect on hepatic metabolism. Volume of distribution may also be increased in elderly patients because of their greater percentage of adipose tissue. Ethnic background can significantly influence both drug metabolism and the pharmacodynamics of a variety of drugs. Thus, the physician should carefully consider patient characteristics when prescribing psychotropic medications and should engage in therapeutic drug monitoring when there is any doubt about the plasma drug levels that a given dosing regimen will achieve. PMID- 8407859 TI - Basic pharmacokinetic principles and their application to psychotropic drugs. AB - The application of pharmacokinetic principles to the administration of psychotropic medications provides a rational approach to understanding factors influencing the time course and intensity of drug action. When plasma concentrations of a given medication can be linked to levels at receptors in the brain, the pharmacodynamic effects of that drug can then be predicted. The pharmacokinetic parameter of most interest to clinicians is clearance, which describes the rate of drug removal per unit of plasma concentration. For any given drug, this parameter varies widely from person to person and may be markedly altered both in elderly patients and in patients with either kidney or liver disease. The clearance of a given medication can also be either increased or reduced by coadministration of other drugs. Such drug:drug interactions can potentially result in either reduced therapeutic effect or toxicity. Elimination half-life is determined by clearance and by another pharmacokinetic parameter, volume of distribution. Elimination half-life will be related to both the time necessary for plasma concentrations to reach steady state with repeated dosing and for the drug to be washed out after it is discontinued. PMID- 8407860 TI - Immunological identification of the major disulfide-linked light component of silk fibroin. AB - The light protein components of silk fibroin were analyzed by Western blotting using polyclonal antibodies against two synthetic peptides that are segment of the light (L-) chain [Yamaguchi et al. (1989) J. Mol. Biol. 210, 127-139] and P25 [Chevillard et al. (1986) Sericologia 26, 435-449], respectively, and against the whole L-chain. Both the L-chain and P25 were present with fibroin in the lumen of the posterior silk gland and in the cocoon. The L-chain was identified as the major light component that was disulfide-linked to the fibroin heavy (H-) chain, whereas P25 was a minor component whose association with the H-chain was suggested to be non-covalent. PMID- 8407861 TI - Purification and structure of rat erythroid-specific delta-aminolevulinate synthase. AB - The existence of erythroid form delta-aminolevulinate synthase (ALAS-E) was historically a matter of some controversy. To obtain direct evidence for a unique ALAS-E, we have purified ALAS-E to homogeneity for the first time, from rat reticulocyte lysate. The papain digestion method was used at the initial step of the purification to overcome the difficulty which repeatedly hampered earlier attempts to purify ALAS-E. The size of the purified papain-resistant core catalytic domain of ALAS-E was estimated electrophoretically to be 49,000 Da. The pH optimum (7.6) and apparent Km values for the substrates, glycine (6.5 mM) and succinyl-CoA (2 microM), were similar to those of the non-specific form of delta aminolevulinate synthase (ALAS-N); but, in contrast to ALAS-N, the substrate inhibition by succinyl-CoA was not evident in ALAS-E. We then isolated cDNA and genomic DNA clones encoding rat ALAS-E. By combining the nucleotide sequence information of the cDNA and genomic clones, the rat ALAS-E precursor is predicted to be composed of 587 amino acids with a calculated molecular mass of 64,841 Da. All the peptide sequences determined directly from the purified protein agreed with those predicted from the nucleotide data, demonstrating the existence of ALAS-E. Analysis of the papain-resistant core domain further revealed that it overlaps with the evolutionally conserved segment that has been noticed by sequence alignment analysis of ALA synthases from various species. PMID- 8407862 TI - Biosynthesis of prenyl diphosphates by cell-free extracts from mammalian tissues. AB - When assayed by the conventional method for prenyltransferase using a combination of [1-14C]isopentenyl and geranyl diphosphates, 100,000 x g supernatants of homogenates of rat liver and brain catalyzed the formation of geranylgeranyl diphosphate at a much lower rate than that of farnesyl diphosphate. Surprisingly, however, the formation of geranylgeranyl diphosphate in incubations of [1 14C]isopentenyl diphosphate alone with these enzyme systems was comparable to that of farnesyl diphosphate. Addition of dimethylallyl diphosphate to the same enzyme systems in the presence of [1-14C]isopentenyl diphosphate resulted in a marked increase in the rate of formation of farnesyl diphosphate, while the rate of formation of geranylgeranyl diphosphate was saturated. Metabolic labeling of rat liver and kidney slices with [5-3H]mevalonic acid revealed that the major prenyl residue of the detectable prenylated proteins was actually the geranylgeranyl group. Coupled with the previous finding that geranylgeranyl diphosphate accumulates during metabolic labeling of rat liver slices with [2 3H]mevalonic acid [Sagami, H., Matsuoka, S., and Ogura, K. (1991) J. Biol. Chem. 266, 3458-3463], these results indicate that the rate of de novo synthesis of geranylgeranyl diphosphate from mevalonic acid is comparable to that of farnesyl diphosphate. PMID- 8407863 TI - Geranylgeranyl diphosphate synthase catalyzing the single condensation between isopentenyl diphosphate and farnesyl diphosphate. AB - Geranylgeranyl diphosphate synthase was purified 191-fold from bovine brain by Mono Q column chromatography followed by preparative isoelectric focusing electrophoresis and Superose 12 gel filtration. The synthase had a pI value at 6.0, and it was made free of farnesyl diphosphate synthase, the pI of which was 5.1. The partially purified enzyme catalyzed the formation of geranylgeranyl diphosphate from isopentenyl diphosphate and farnesyl diphosphate with the Km values for isopentenyl diphosphate and farnesyl diphosphate being 14 and 0.8 microM, respectively. Dimethylallyl diphosphate and geranyl diphosphate were poor substrates with velocities of only 0.003 and 0.03, respectively, relative to that of farnesyl diphosphate. These results indicate that geranylgeranyl diphosphate synthase catalyzes a single condensation between isopentenyl diphosphate and farnesyl diphosphate and that farnesyl diphosphate is the common intermediate at the branch point for the synthesis of geranylgeranylated proteins as well as cholesterol, ubiquinone, dolichol, and farnesylated proteins. The enzyme required Mg2+ or Mn2+ for maximum activity. Octylglucoside showed a stimulatory effect on the enzyme activity. PMID- 8407864 TI - Both medullasin and human leukocyte elastase are essentially devoid of elastinolytic activity. AB - Elastinolytic activity of medullasin was investigated precisely and compared with that of human leukocyte elastase, because the structure of medullasin is quite similar to that of human neutrophil elastase, which was reported to have elastinolytic activity. When elastinolytic activity of medullasin and human leukocyte elastase was determined by employing unstained elastin fibers and measuring the increase in 280-nm absorbance of the supernatant, elastinolytic activity amounting to several percent of that of porcine pancreas elastase was apparently observed. However, the susceptibility of elastin preparations to these proteases was proportional to their hydroxyproline content. Both medullasin and human leukocyte elastase digested collagen fibers obtained from bovine Achilles tendon to the same extent as collagenase from Clostridium histolyticum. When elastinolytic activity was determined by employing elastin fibers stained with orcein, both proteases showed negligible elastinolytic activity. The activity remained negligible even when the pH or ionic strength of the reaction mixture was altered. These results indicate that medullasin and human leukocyte elastase are essentially devoid of elastinolytic activity, and that apparent elastinolytic activity observed when unstained elastin fibers were employed as the substrate is due to the digestion of collagen fibers mingled with elastin preparations. PMID- 8407865 TI - Heterogeneity of dystrophin-associated proteins. AB - The proteins which compose the complex of dystrophin and its associated proteins were analyzed by two-dimensional PAGE, i.e., electrofocusing in the presence of 8 M urea followed by SDS PAGE. Silver-staining of the gel showed many more spots than were expected from the results of one-dimensional SDS PAGE. By examination of their reactivity with specific antibodies, various lectins and 3 (trifluoromethyl)-3-(m-[125I]iodophenyl)-diazirine, most of these spots were identified as dystrophin and its associated proteins described previously. Several as yet unidentified minor proteins were also detected. Dystrophin associated protein A1 was separated into two groups, alpha-A1 and beta-A1, both composed of numerous spots. These groups differed from each other in isoelectric point, molecular mass, and reactivity with antibodies. The beta-A1 group (64 kDa) was more basic than the alpha-A1 group (60 kDa). Beta-A1 but not alpha-A1 reacted with several lectins, indicating that beta-A1 is a glycoprotein. This is incompatible with the report that 59DAG, which corresponds to A1 (alpha-A1 + beta A1), is not a glycoprotein [Ervasti et al. (1991) Cell 66, 1121-1131]. The charge heterogeneity observed in alpha-A1 and beta-A1 may be partially due to differential phosphorylation. The charge heterogeneity of A2 and A3a may, at least to some extent, be due to differential sialilation of their carbohydrates. PMID- 8407866 TI - Purification of the bovine nicotinic acetylcholine receptor alpha-subunit expressed in baculovirus-infected insect cells. AB - The bovine skeletal muscle nicotinic acetylcholine receptor alpha-subunit was produced in insect, Spodoptera frugiperda (Sf9), cells by infection with a recombinant baculovirus. The expressed alpha-subunit protein could not be solubilized efficiently with Triton X-100 or sodium cholate, but could be solubilized efficiently with Zwittergent 3-14, sodium dodecyl sulfate or tris(hydroxymethyl)aminomethane dodecylsulfate. After solubilization of the alpha subunit protein with Zwittergent 3-14 from the Triton X-100-insoluble fraction, the alpha-subunit protein was purified by concanavalin A-Sepharose chromatography and DEAE ion-exchange chromatography. A milligram quantity of the alpha-subunit protein could be purified from 8 g (wet weight) of Sf9 cells infected with the recombinant baculovirus. Chromatographic analyses including hydroxyapatite chromatography, DEAE ion-exchange chromatography, gel filtration chromatography and chromatofocusing and sucrose density gradient centrifugation analysis suggest that the purified alpha-subunit protein is homogeneous. The purified alpha subunit protein had a high affinity for 125I-alpha-bungarotoxin and was glycosylated with a high mannose-type N-linked oligosaccharide side chain. These results indicate that purification of ion channel proteins produced by the baculovirus expression system is a promising approach to structural analysis of ion channel proteins, which are extremely rare membrane proteins in native tissues. PMID- 8407867 TI - Estimation of the possible recognition sites for thrombomodulin, procoagulant, and anticoagulant proteins around the active center of alpha-thrombin. AB - alpha-Thrombin has several characteristic module structures around its active center. Previously we showed that a synthetic peptide, TWTANVGKGQPS, corresponding to the residues Thr147 to Ser158 of the B-chain of human thrombin, a possible interaction site of thrombomodulin near the active center of thrombin, specifically blocked the interactions between thrombin and thrombomodulin, fibrinogen, Factor V, or platelets [Suzuki, K. & Nishioka, J. (1991) J. Biol. Chem. 266, 18498-18501]. To elucidate further the role of the other module structures, we studied the effects of several synthetic peptides; FRKSPQELL, LLYPPWDKNF, RIGKHSRTRYER, LEKIYIHP, RYNWREN, DSTRIRI, EGDSGGP, and SWGEGCDRDGK, respectively corresponding to the residues Phe19 to Leu27, Leu45 to Phe54, Arg62 to Arg73, Leu81 to Pro88, Arg89 to Asn95, Asp175 to Ile181, Glu202 to Pro208, and Ser226 to Lys236 of the B-chain of human thrombin, which are located around the active center, as well as TWTANVGKGQPS, on the interaction between thrombin and thrombomodulin, protein C, fibrinogen, Factor V, antithrombin III, or hirudin. Thrombin-thrombomodulin interaction was inhibited significantly by RYNWREN as well as TWTANVGKGQPS, and partially by LLYPPWDKNF. The inhibitory effects of the two former peptides were additive and thrombomodulin directly bound to them. RYNWREN and TWTANVGKGQPS also increased the Km values 3-7 times for protein C as compared with the conditions without peptide. Thrombin-induced protein C activation in the absence of thrombomodulin was specifically blocked by EGDSGGP. Thrombin-induced fibrinogen clotting was blocked by FRKSPQELL, RIGKHSRTRYER as well as TWTANVGKGQPS at lower concentrations, and by RYNWREN and DSTRIRI at higher concentrations. Thrombin-induced Factor V activation was blocked by FRKSPQELL, RIGKHSRTRYER as well as TWTANVGKGQPS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407868 TI - Cloning of a human acid sphingomyelinase cDNA with a new mutation that renders the enzyme inactive. AB - A cDNA encoding human acid sphingomyelinase was initially obtained by screening a placental cDNA library in lambda gt11 with a synthetic oligonucleotide probe and subsequently with partial cDNA. The full-length cDNA, hPSM55, comprised 2,376 nucleotides, with a 5' untranslated sequence of 122 nucleotides, an open reading frame of 1,884 nucleotides encoding a protein of 627 amino acids, and a 3' untranslated region of 370 bases. hPSM55 was almost identical to pASM-1FL reported by Schuchman et al. (J. Biol. Chem. 266, 8531-8539, 1991) except for a 6 base pair deletion in the signal peptide, which indicated the possible removal of valine and leucine residues between positions 36 and 37, and a 463T- to C transition, which indicated a possible substitution of 155arginine for cystine. This cDNA was expressed in both COS-7 cells and Chinese hamster ovary cells. There was no increase in acid sphingomyelinase activity in either cell line following transfection. However, the correction of a single base change, 463C to T, in hPSM55 caused increased acid sphingomyelinase activity in transfectants. These results suggest that the mutation of nucleotide 463C to T plays an important role in the catalytic activity of acid sphingomyelinase. PMID- 8407869 TI - Characterization of poly C preferential ribonuclease from chicken liver. AB - Poly C preferential RNase previously reported by Levy and Karpetsky [J. Biol. Chem. 255, 2153-2159 (1980)] and Miura et al. [Chem. Pharm. Bull. 32, 4053-4060 (1984)] was extensively purified from chicken liver to homogeneity as determined by SDS-PAGE (RNase CL2). The poly C preference over poly U was slightly higher than that of bovine pancreatic RNase A. However, the kinetic constants for 8 dinucleoside phosphates, CpY and UpY (Y = one of A, G, U, and C) as substrates showed that RNase CL2 was preferential for cytidylic acid, but less so than RNase A, and the influence of Y base on the rate of hydrolysis of CpY or UpY was less marked than in the case of RNase A. The primary structure of RNase CL2 was determined. The molecular weight calculated from the sequence was 13,420. Comparison of the amino acid sequence of RNase CL2 with those of other vertebrate RNases showed that RNase CL2 is a member of the RNase A family, but is not a non secretory RNase. It retains 3 disulfide bridges of RNase A, but Cys65-Cys72 of RNase A is missing. As for the active site, the amino acid residues of the P0 and P1 sites of RNase A are completely conserved. Among the B1 site components, Thr45 (RNase A numbering) is conserved, but Phe120 and Ser123 are substituted by Leu and Thr, respectively. Among the B2 site residues, Gln69, Asn71, and Glu111 are substituted by other amino acids. PMID- 8407870 TI - High concentration of glucitol in fetal serum and glucitol permeable cells. AB - We found that the glucitol concentration was extraordinarily high in bovine fetal serum, which is routinely used for cell culture in laboratories: it was as much as two orders of magnitude higher than that reported for human adult serum. We also confirmed that the serum glucitol concentration in new born babies was on average 5.5-fold higher than in the maternal serum. These observations raise the possibility that some tissue(s) demand extracellular glucitol during embryogenesis and this indicates that the cells in such tissues could be permeable to glucitol. Since the hepatic metabolism of glucitol had been reported, we investigated glucitol permeability and its metabolism in rat hepatoma cells, Reuber H-35. The cells rapidly incorporated glucitol but the mode of incorporation was unusual: the incorporation rate was still proportional to the ambient glucitol concentration at 100 mM. The major part of the incorporated glucitol underwent metabolic conversion to probably negatively charged metabolites. Active synthesis of glucitol was also observed in the same cells. The cells proliferated normally in medium containing glucitol instead of glucose. This observation may indicate that glucitol can substitute for glucose in the culture of H-35 cells. PMID- 8407871 TI - Unitary distance of actin-myosin sliding studied using an in vitro force-movement assay system combined with ATP iontophoresis. AB - To obtain information about the mechanism of ATP-dependent actin-myosin sliding responsible for muscle contraction, we studied the "unitary" distance of sliding between a myosin-coated glass microneedle and actin filament arrays (actin cables) in a giant algal cell induced by iontophoretic application of ATP, attention being focused on the minimum distance of ATP-induced sliding when the amount of applied ATP was gradually decreased in the presence of hexokinase and D glucose. The number of myosin heads interacting with actin cables was reduced to less than 100, as judged from the maximal force Po (approximately 100 pN) generated by myosin heads on the needle in the presence of 2 mM ATP. When the amount of iontophoretically applied ATP was decreased by reducing the amount of charge passed through the ATP electrode from 80 to 2 nC, the distance of ATP induced actin-myosin sliding decreased almost linearly from approximately 100 to approximately 10 nm, no detectable actin-myosin sliding being observed with further reduction of the charge passed through the electrode. The amount of external load exerted by the bent microneedle was less than 1% of Po for the sliding distance < 50 nm. The actin-myosin sliding distances with a small amount of ATP slightly above the amount required to induce the minimum sliding distance were distributed around integral multiples of 10 nm, suggesting that the unitary distance of actin-myosin sliding coupled with ATP hydrolysis is of the order of 10 nm. PMID- 8407872 TI - Preferential hydrolysis of phosphatidylethanolamine in rat ischemic heart homogenates during in vitro incubation. AB - Phospholipid-hydrolyzing activities were examined in rat hearts with ischemia induced by occlusion of the left main coronary artery. When homogenates of ischemic heart were incubated in vitro at 37 degrees C, a significant amount of phosphatidylethanolamine (PE) was degraded, whereas the contents of other phospholipids did not change significantly. During the incubation, a stoichiometrical amount of lysoPE was concomitantly formed. The lysoPE formed had mainly saturated fatty acids and its composition resembled that of fatty acids detected at the sn-1 position in the glycerol backbone of heart PE. No appreciable PE degradation was observed in homogenates prepared from nonischemic rat heart. No difference in phospholipase activities was found between ischemic and nonischemic heart homogenates when exogenous radioactive phospholipids were used as substrates. Rabbit anti-rat 14-kDa type II phospholipase A2 antibody suppressed the degradation of PE observed in ischemic heart homogenates. These findings indicate that the type II phospholipase A2 activity may be involved in the breakdown of endogenous PE in ischemic heart homogenates. PMID- 8407873 TI - Flagellar growth in a filament-less Salmonella fliD mutant supplemented with purified hook-associated protein 2. AB - Bacterial flagellum consists of a basal body, a hook, HAP1 (hook-associated protein 1), HAP3, a long helical filament, and a cap (composed of HAP2), all connected in series. The mutant deficient in the HAP2 structural gene (fliD) of Salmonella typhimurium has flagella composed of only hook-HAP1-HAP3 and excretes flagellin monomers into the culture medium. However, when purified HAP2 was added to this mutant, the flagellin stopped leaking out and flagellar filaments grew. Turnover of HAP2 was not necessary for the growth of a filament. Therefore HAP2 facilitates the polymerization of endogenous flagellin, apparently without falling off the filament tip. This experimental system with exogenous HAP2 allowed us to synchronize filament growth; the average rate of filament growth can be estimated by measuring the length of grown filaments at various time periods in electron micrographs. The initial growth rate was about 30 nm/min, which corresponds to one flagellin per second. PMID- 8407874 TI - Purification and characterization of homospermidine synthase in Acinetobacter tartarogenes ATCC 31105. AB - Homospermidine synthase, catalyzing the formation of homospermidine [H2N(CH2)4NH (CH2)4NH2] from putrescine and NAD+ with concomitant liberation of NH3, was purified 600-fold over the crude extract with a yield of about 14% to homogeneity from Acinetobacter tartarogenes ATCC 31105. The enzyme had a native molecular mass of 102 kDa, with a pI of 5.0, and was apparently composed of two identical subunits (52 kDa), suggesting that a single protein catalyzes two serial reactions, oxidation of putrescine to 4-aminobutyraldehyde and subsequent reduction of the putative Schiff base formed between this aldehyde and a second molecule of putrescine to homospermidine. The Km values for putrescine and NAD+ were 280 and 18 microM, respectively. 1,3-Diaminopropane and cadaverine were inactive as substrates, and NAD+ could not be replaced by NADP+. 1,3 Diaminopropane and NADH were potent competitive inhibitors. The enzyme had a pH optimum of 8.7, was most active at 30 degrees C, and required K+ and dithiothreitol for full activity. Putrescine and NAD+ protected the enzyme from the inhibition by thiol reagents. The NH2-terminal amino acid sequence was AQWPVYGKISGPVVI. Some of these properties were compared with those of the homospermidine synthases from a photosynthetic bacterium, Rhodopseudomonas viridis and a plant, Lathyrus sativus. PMID- 8407875 TI - Generation of a monoclonal antibody specific for ganglioside GM4: evidence for GM4 expression on astrocytes in chicken cerebellum. AB - We established a murine monoclonal antibody (MAb) specific for ganglioside GM4 by immunizing C3H/HeN mice with chemically synthesized GM4 adsorbed to Salmonella minnesota, followed by fusion with mouse myeloma cells. The MAb, designated as AMR10, was shown to exhibit high binding specificity, reacting only with the ganglioside GM4 used for immunization and native GM4 from human brain. We determined the distribution of GM4 in adult chicken cerebellum by means of the immunofluorescence technique with the MAb. Our study revealed that GM4 expression was associated with astrocytes in the granular layer and the white matter, but not with myelin in any layers of the chicken cerebellar cortex. PMID- 8407876 TI - Alterations in activities of protein phosphatases PP1 and PP2A in T and B lymphocytes of autoimmune MRL/MpJ-lpr/lpr mice. AB - Activities of protein phosphatases PP1 and PP2A were determined in T and B lymphocytes of autoimmune-prone MRL/MpJ-lpr/lpr mice (MRL/lpr mice) and two control strains, MRL/MpJ-(+)/+ mice (MRL/+/+ mice) and C3H/HeJ mice. Potential PP1 activity, which was measured after treatment of cell extract with Co(2+) trypsin, was much higher in T lymphocytes than B lymphocytes. However, no difference in the activity was observed between MRL/lpr mice and the controls. Spontaneous PP2A activity showed similar levels in T and B lymphocytes from normal mice, but potential PP2A activity, which was measured after treatment with 2-mercaptoethanol, was significantly higher in T lymphocytes from MRL/lpr mice than those from controls. No differences were detected in PP1 or PP2A activities in B lymphocytes. From these results, our previous data [Matsuzawa, S. et al. (1992) J. Biochem. 111, 472-477] demonstrating increases in potential activities of PP1 and PP2A in lymphoid tissues from autoimmune MRL/lpr mice can be interpreted as follows. 1) The increase in potential PP1 activity of the lymphoid tissues from MRL/lpr mice is caused by replacement of B lymphocytes by abnormal T lymphocytes, which accumulate in enormous numbers. 2) The increase of potential PP2A activity in the lymphoid tissues from MRL/lpr mice is caused by the increase in this activity in their T lymphocytes. PMID- 8407877 TI - Purification and characterization of the arylphorin gene specific binding protein from an embryonic cell line of Sarcophaga peregrina (flesh fly). AB - Previously, we purified a DNA binding protein from a nuclear extract of the fat body of Sarcophaga peregrina larvae that binds to the ACCACAACA motif in the 5' upstream region of the arylphorin gene, and suggested that this protein is a transcriptional activator of the arylphorin gene. In this study, we detected and purified the same protein (ABP-1) from an embryonic cell line of Sarcophaga that does not express the arylphorin gene. Unlike the fat body, which synthesizes arylphorin actively, the embryonic cells were found to contain an additional DNA binding protein (ABP-2) that bound to the same DNA probe as ABP-1, suggesting a novel mechanism of regulation of the arylphorin gene. PMID- 8407878 TI - Molecular cloning, nucleotide sequencing, and affinity labeling of bovine liver UDP-glucose pyrophosphorylase. AB - A bovine liver cDNA encoding UDP-glucose pyrophosphorylase [EC 2.7.7.9], which catalyzes the reversible uridylyl transfer between glucose 1-phosphate and MgUTP, has been cloned by the use of oligonucleotide probes synthesized on the basis of partial amino acid sequences of the enzyme. The cDNA clone contained a 1,689 base pair insert including the complete message for the subunit polypeptide (508 amino acid residues) of the octameric enzyme. The bovine liver enzyme shows significant sequence similarities with the enzymes from potato tuber and a slime mold, Dictyostelium discoideum, but not with the enzyme from Escherichia coli, or ADP glucose pyrophosphorylases from rice seed and E. coli. To probe the substrate binding site in the bovine liver enzyme, the purified enzyme was incubated with an affinity labeling reagent, uridine triphosphopyridoxal, and then reduced with sodium borohydride. The enzyme was inactivated rapidly and irreversibly by the reagent at low concentrations. The inactivation was almost completely retarded by UDP-glucose and MgUTP. Structural analysis of the labeled enzyme revealed that three lysyl residues, Lys291, Lys357, and Lys396, were modified by the reagent. The three lysyl residues are conserved at the corresponding positions in the sequence of the potato tuber enzyme, in which they have catalytically important functions. These results show that the active-site structure of bovine liver UDP glucose pyrophosphorylase is very similar to that of the potato tuber enzyme. PMID- 8407879 TI - Site-directed mutagenesis of a hexapeptide segment involved in substrate recognition of phenylalanine dehydrogenase from Thermoactinomyces intermedius. AB - Phenylalanine dehydrogenase from Thermoactinomyces intermedius and leucine dehydrogenase from Bacillus stearothermophilus show a 59% sequence similarity in their substrate-binding domains, although their substrate specificities are different. We prepared a phenylalanine dehydrogenase mutant enzyme whose inherent hexapeptide segment (124F-V-H-A-A-129R) in the substrate-binding domain was replaced by the corresponding part of leucine dehydrogenase (M-D-I-I-Y-Q) in order to investigate the mechanism of substrate recognition by phenylalanine dehydrogenase. The catalytic efficiencies (kcat/Km) of the mutant enzyme with aliphatic amino acids and aliphatic keto acids as substrates were 0.5 to 2% of those of the wild-type enzyme. In contrast, the efficiencies for L-phenylalanine and phenylpyruvate decreased to 0.008 and 0.035% of those of the wild-type enzyme, respectively. These results suggest that the hexapeptide segment plays an important role in the substrate recognition by phenylalanine dehydrogenase. PMID- 8407880 TI - Structural study of the N-linked oligosaccharides of hepatocyte growth factor by two-dimensional sugar mapping. AB - The structures of the N-linked oligosaccharides on recombinant human hepatocyte growth factor (rh-HGF) expressed by Chinese hamster ovary (CHO) cells were studied by two-dimensional sugar mapping. The oligosaccharides released from the glycopeptides by peptide: N-glycosidase F (PNGase F) treatment were tagged with 2 aminopyridine at the reducing ends. The alpha-chain was linked by biantennary, triantennary, and tetraantennary oligosaccharides, but the dominant oligosaccharides linking the beta-chain were biantennary (> 85%). There was no significant difference in oligosaccharide structures between the two glycosylation sites on each chain, that is, Asn263 and Asn371 on the alpha-chain, and Asn535 and Asn622 on the beta-chain. The linkage of sialic acid to the non reducing terminal galactose was identified as NeuAc alpha(2-3) by 1H-NMR spectrometry. The structures of the N-linked oligosaccharides from rat HGF were also studied. Triantennary oligosaccharides were obtained from the alpha-chain and a biantennary oligosaccharide was obtained from the beta-chain. This result indicates that the alpha-chain is also linked by higher branched oligosaccharides than the beta-chain in rat HGF. PMID- 8407881 TI - Immature human chorionic gonadotropin (hCG) in first trimester placental cells is bound to an ATP-binding protein forming high-molecular-weight hCG. AB - Human chorionic gonadotropin (hCG) in first trimester placental cells is made up of immature alpha- and beta-subunits containing only N-linked high-mannose sugar chains, which are of 21 kDa for the alpha-subunit and 23 and 19 kDa for the beta subunit. However, the apparent molecular weight of immature hCG from placental cell extracts has been estimated from gel filtration to be much higher (100-200 kDa; high molecular weight-hCG, HMW-hCG) based on gel filtration than the theoretical value (approximately 44 kDa) of the alpha beta dimer (alpha beta hCG). We prepared a gel-filtered fraction containing HMW-hCG and investigated treatments for converting it to alpha beta-hCG. We found that the molecular weight of HMW-hCG was decreased to close to that of alpha beta-hCG by treatment with acetone, proteases, or chelating agents. These treatments also shifted the isoelectric point of HMW-hCG from the acidic region (pI = 4-6) to the alkaline (pI = 9-11), approximating to that of alpha beta-hCG. We also found that HMW-hCG, but not acetone-treated HMW-hCG, bound to ATP-agarose resin. These results suggested that the immature alpha beta-hCG molecule in placental cells may be bound to an acidic ATP-binding protein to form HMW-hCG. PMID- 8407882 TI - The molecular features and catalytic activity of CuA-containing aco3-type cytochrome c oxidase from a facultative alkalophilic Bacillus. AB - Cytochrome aco3 of Bacillus YN-2000 was purified by an improved procedure and its molecular features and catalytic activity were extensively studied. The enzyme molecule was composed of three subunits with M(r)s of 50,000, 41,000, and 22,000, and contains 1 molecule each of cytochrome a, cytochrome c, and cytochrome o3 in the minimal structural unit (M(r), 113,000). The 41,000 subunit (subunit II) contains heme c. The EPR, optical, and resonance Raman spectra of the oxidized enzyme demonstrated the presence of CuA whose coordination environment bore close resemblance to that of the aa3-type cytochrome c oxidase. Resonance Raman studies demonstrated that the cytochrome a moiety was similar to that of an aa3-type oxidase and also that the cytochrome o3 contained a five-coordinated high-spin heme with histidine as an axial ligand. The Fe-CO stretching mode of the cytochrome o3.CO complex was observed at 520 cm-1, which is the same frequency as that of cytochrome aa3.CO. The oxygen consumption activity of cytochrome aco3 was measured using several kinds of cytochromes c as the electron mediators. The reaction between cytochrome aco3 and eucaryotic cytochromes c was completely inhibited by poly-L-lysine. In contrast, poly-L-lysine was indispensable for sufficient reaction between the oxidase and Bacillus YN-2000 cytochrome c-553, the physiological electron donor. The combined results on the structure and enzymatic properties suggest that the cytochrome aco3 is very similar to cytochrome caa3 except that the cytochrome aco3 has cytochrome o3 in place of cytochrome a3 and the cytochrome c component has a very low redox midpoint potential (95 mV).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407883 TI - Thymidine phosphorylase activity associated with platelet-derived endothelial cell growth factor. AB - Partial complementary DNA (cDNA) for thymidine phosphorylase (dThdPase) was cloned by means of a polymerase chain reaction. There was complete sequence identity between the amino acid sequence deduced from the nucleotide sequence of a clone (288 nucleotides) and the residues of platelet-derived endothelial cell growth factor (PD-ECGF). The amino acid sequence of all four peptide fragments from purified human dThdPase could be aligned with that of PD-ECGF. Our data indicate that residues 125-244 of PD-ECGF are identical to the sequence of human dThdPase. The molecular weights of human dThdPase and recombinant PD-ECGF (rPD ECGF) that lacks 10 amino acids at the amino terminal were 55 and 52 kDa, respectively. Anti-PD-ECGF antibody recognized dThdPase, and anti-dThdPase antibody recognized rPD-ECGF. rPD-ECGF had dThdPase activity and its specific activity was similar to that of purified human dThdPase. dThdPase activity and molecules were detected in COS cells transfected with human PD-ECGF cDNA, but not in nontransfected cells. The sizes of PD-ECGF and dThdPase in the transfected COS cells were identical. These data suggest that human dThdPase is identical to PD ECGF. PMID- 8407884 TI - Cell density-dependent regulation of hepatocyte growth factor receptor on adult rat hepatocytes in primary culture. AB - The proliferative potential of hepatocytes in the normal intact liver is highly suppressed, but they proliferate actively after liver injury. In this study, adult rat hepatocytes in primary culture were used to study mechanisms controlling hepatocyte growth in liver regeneration. DNA synthesis in hepatocytes cultured at a low cell density was highly stimulated in response to hepatocyte growth factor (HGF), but this stimulatory effect was not so obvious in hepatocytes cultured at a high cell density. In close parallel to the potency of DNA synthesis, the amounts of 125I-HGF specifically bound to hepatocytes cultured at a low cell density were much greater than in high cell density culture. Scatchard plots revealed that change in the specific binding of 125I-HGF was due to change in the number of high-affinity HGF receptors, but without change in the Kd values. Affinity cross-linking of the HGF receptor with 125I-HGF confirmed the higher expression of HGF receptor on hepatocytes cultured at a lower cell density. Since there was no significant change in the expression of c-met mRNA in hepatocytes cultured at different cell densities, the number of cell surface HGF receptors is probably regulated by post-transcriptional mechanisms. We also found that the rates of HGF-induced down-regulation and recovery of HGF receptors on hepatocytes cultured at a low cell density were much faster than in cases of a high cell density.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8407885 TI - A helix-loop-helix transcription factor-like gene is located at the mi locus. AB - The mouse microphthalmia phenotype is complex and consists of one or more of the following phenotypic alterations: a lack of pigmentation, small eyes, a mast cell defect, and bone abnormalities. The locus for this allele has been assigned to chromosome 6. A single gene defect that produces such a pleiotropic effect has suggested some involvement at a control point in development. Recently a mutant line of mice carrying a transgene insertion, which represents a new allelic form of mi, was described. The integration site of the transgene from these mi(tg) mice was cloned and analyzed. An exon sequence was discovered adjacent to the insertion. Computer analysis of this nucleotide sequence revealed the presence of a motif indicative of the helix-loop-helix class of transcription factors. The gene was expressed in a number of tissues from wild type animals but was absent in the tissue RNA from mi(tg) mice. Southern blot analysis demonstrated a deletion of some of the genetic material for this gene in the mi(tg) mice. This is consistent with the lack of expression in the mi(tg) mice. Interestingly, when DNA from other mi allelic variants was subjected to a similar analysis, a deletion was also observed in this gene in two other mi lines. Taken together, these data suggest that the gene encoding this new helix-loop-helix DNA-binding protein, and residing in the mi locus, is a strong candidate for the mi gene. PMID- 8407886 TI - Molecular piracy of mammalian interleukin-8 receptor type B by herpesvirus saimiri. AB - Viruses are known to acquire and modify the genes of their hosts to attain a survival advantage in the host environment. Herpesvirus saimiri (HVS) is a T lymphotropic virus that causes fatal lymphoproliferative diseases in several non human primates. The gene ECRF3 of HVS was most likely acquired from a primate host. ECRF3 encodes a putative seven-transmembrane-domain receptor that is remotely related (approximately 30% amino acid identity) to the known mammalian alpha and beta chemokine receptors, namely interleukin-8 receptor (IL8R) types A and B and the MIP-1 alpha/RANTES receptor, respectively. Chemokines regulate the trafficking, activation, and, in some cases, proliferation of myeloid and lymphoid cell types. We now show that ECRF3 encodes a functional receptor for the alpha chemokines IL-8, GRO/melanoma growth stimulatory activity (MGSA), and NAP-2 but not for beta chemokines, a specificity identical to that of IL8RB. Paradoxically, IL8RA shares 77% amino acid identity with IL8RB but is not a receptor for GRO/MGSA or NAP-2. This is the first functional characterization of a viral seven-transmembrane-domain receptor. It suggests a novel role for alpha chemokines in the pathogenesis of HVS infection by transmembrane signaling via the product of ECRF3. PMID- 8407887 TI - Potent inhibition of human tumor p21ras farnesyltransferase by A1A2-lacking p21ras CA1A2X peptidomimetics. AB - The ras oncogene product p21ras requires farnesylation and subsequent plasma membrane association for its transforming activity. This key post-translational modification is catalyzed by p21ras farnesyltransferase, which transfers farnesyl from farnesylpyrophosphate to the cysteine of the CA1A2X carboxyl-terminal tetrapeptide of p21ras. In the present report, we describe potent inhibition of p21ras farnesyltransferase by CA1A2X peptidomimetics containing no peptidic amide bonds. We synthesized a series of CA1A2X analogues where the 2 aliphatic amino acids A1 and A2 were replaced by a hydrophobic spacer, 3-aminomethylbenzoic acid (AMBA). The peptidomimetic Cys-AMBA-Met, inhibits p21ras farnesyltransferase from human colon carcinoma (COLO-205) and Burkitt's lymphoma (Daudi) with IC50 values of 60 and 120 nM, respectively. Cys-AMBA-Met is 3-, 8-, and 9-fold (COLO-205) and 2-, 5-, and 7-fold (Daudi) more potent than the corresponding tetrapeptides of p21KB-ras (CVIM), p21N-ras (CVVM), and p21KA-ras (CIIM), respectively. Replacing methionine at the X position with negatively charged glutamate reduces its ability to inhibit the enzyme, whereas positively charged lysine at this position abolishes the inhibitory character of the peptidomimetic. A hydrophobic moiety at the X position, as in Cys-AMBA-Phe, retains potent inhibitory activity. Leucine in the X position of CA1A2X is a post-translational signal for protein geranylgeranylation rather than farnesylation, and, as expected, Cys-AMBA-Leu does not inhibit the enzyme. Furthermore, CVIM, CVVM, and CIIM are farnesylated by human p21ras farnesyltransferases and inhibit these enzymes by serving as alternative substrates. In contrast, the peptidomimetics described here are true p21ras farnesyltransferase inhibitors since none is farnesylated by this enzyme. PMID- 8407888 TI - Preferential binding of histone H1 to four-way helical junction DNA. AB - Histone H1 is a major chromatin protein, which stabilizes the nucleosome, has an essential role in organizing nucleosomes into higher order structures, and may have a role as a repressor of transcription (van Holde, K. E. (1989) Chromatin, Springer Publishing Co., New York). Here we show that H1 forms a defined complex with a synthetic four-way junction of DNA strands even in the presence of an excess of linear nonspecific competitor DNA. The four-way junction also competes efficiently against two duplex DNA molecules, which together have the same sequence information as the four-way junction molecule. Another major chromatin protein, high mobility group protein 1, also binds four-way junction structures specifically (Bianchi, M. E., Beltrame, M., and Paonessa, G. (1989) Science 243, 1056-1059), and this similar behavior may indicate a related function of these proteins. Our finding may suggest that four-way DNA junction is a structural equivalent to the main H1 binding site in the nucleosome: a crossover of double helical DNA at the point where the DNA enters and exits the nucleosome (Allan, J., Hartman, P. G., Crane-Robinson, C., and Aviles, F. X. (1980) Nature 288, 675 679). PMID- 8407889 TI - Stimulation of tyrosine phosphorylation and accumulation of GTP-bound p21ras upon antibody-mediated alpha 2 beta 1 integrin activation in T-lymphoblastic cells. AB - Integrins are a family of heterodimeric integral plasma membrane proteins that behave as receptors for components of the extracellular matrix and also mediate cell to cell adhesion. Occupation of integrins can result in the transduction of intracellular signals, leading to cytoskeletal reorganization, tyrosine phosphorylation, and induction of gene expression. We report here that the ligation of alpha 2 beta 1 integrin by collagen-adhesion stimulatory anti-alpha 2 and anti-beta 1 antibodies resulted in the accumulation of p21ras in the active GTP-bound state in Jurkat T-lymphoblastoid cells. The activation was accompanied by the tyrosine phosphorylation of proteins of 47-52 kDa. This stimulation of tyrosine phosphorylation and p21ras activation was specific for the activating antibodies and occurred within 2 min of the addition of these antibodies. Although treatment of the cells with the protein kinase C activator, phorbol 12 myristate 13-acetate also resulted in an induction of both cell attachment to collagen and of p21ras activation, tyrosine phosphorylation was not observed. These results demonstrate that alpha 2 beta 1 integrin activation can result in the specific stimulation of tyrosine phosphorylation of 47-52-kDa proteins, as well as activation of a signaling pathway involving p21ras. PMID- 8407890 TI - Biosynthesis of heparin/heparan sulfate. Identification of a 70-kDa protein catalyzing both the D-glucuronosyl- and the N-acetyl-D-glucosaminyltransferase reactions. AB - The D-glucuronosyl- (GlcA) and N-acetyl-D-glucosaminyl- (GlcNAc) transferase reactions involved in heparin/heparan sulfate biosynthesis were assayed, measuring transfer of radiolabeled GlcA or GlcNAc monosaccharide units from the corresponding UDP-sugars to the appropriate oligosaccharide acceptors. The assays were applied to enzyme purification from bovine serum. The two activities remained inseparable through a series of different chromatographic steps, resulting in approximately -2000-fold purification. Further purification was achieved by chromatofocusing, which showed an isoelectric point of pH approximately -7.0, similar for both activities. SDS-polyacrylamide gel electrophoresis (PAGE) of subfractions from the chromatofocusing procedure revealed an approximately 70-kDa protein in amounts reflecting enzyme activity. SDS-PAGE followed by extraction of gel segments and renaturation of proteins showed that the GlcA- and GlcNAc-transferase activities were both recovered from the same single segment, corresponding to the 70-kDa component. It is proposed that the two glycosyltransferase reactions are catalyzed by the same Golgi enzyme (see also Lidholt, K., Weinke, J. L., Kiser, C. S., Lugemwa, F. N., Bame, K. J., Cheifetz, S., Massague, J., Lindahl, U., and Esko, J. D. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 2267-2271). PMID- 8407891 TI - The p27 catalytic subunit of the apolipoprotein B mRNA editing enzyme is a cytidine deaminase. AB - The messenger RNA for apolipoprotein B undergoes a discrete and specific C to U editing of nucleotide 6666. This generates a stop translation codon and defines the carboxyl terminus of apolipoprotein B48. A 27-kDa rat intestinal protein that does not itself edit apolipoprotein B mRNA, but confers editing activity on chick intestinal extracts that do not have intrinsic editing activity, has recently been identified and its cDNA cloned (Teng, B., Burant, C. F., and Davidson, N. O. (1993) Science 260, 1816-1819). Here we show that p27 is homologous in the zinc coordinating region of the active site to cytidine deaminases from Escherichia coli, Bacillus subtilis, yeast, and man and to deoxycytidylate deaminases from T2 and T4 bacteriophages and man. p27 expressed in Xenopus laevis oocyte extracts has cytidine deaminase activity and specifically confers editing activity on chick intestinal extracts. The homologous E. coli cytidine deaminase does not confer editing activity. The zinc-specific chelating agent o-phenanthroline abolishes p27 activity and site-specific apolipoprotein B mRNA editing in rat enterocyte editing extracts. We conclude that p27 is the catalytic subunit of the apolipoprotein B mRNA editing enzyme and is a zinc-containing cytidine deaminase. PMID- 8407892 TI - Arachidonic acid release from aortic smooth muscle cells induced by [Arg8]vasopressin is largely mediated by calcium-independent phospholipase A2. AB - To identify the phospholipase mediating the majority of [Arg8]vasopressin (AVP) induced release of arachidonic acid in A-10 smooth muscle cells, we exploited the specificity inherent in the mechanism-based inhibitor, (E)-6 (bromomethylene)tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one (HELSS), which possesses a 1,000-fold selectivity for inhibition of calcium-independent versus calcium-dependent phospholipases A2. Utilizing [3H]arachidonic acid-labeled A-10 smooth muscle cells, one-half of AVP-inducible [3H]arachidonic acid release was inhibited by pretreatment with only 1 microM HELSS and two-thirds of AVP stimulated [3H]arachidonic acid release was inhibited by 5 microM HELSS. The inhibition of [3H]arachidonic acid release by HELSS was saturable (i.e. no additional inhibition of [3H]arachidonic acid release was present at 10 microM HELSS), specific (i.e. the activities of six intracellular enzymes, as well as the rate of glucose oxidation, were not altered by HELSS treatment), and nontoxic (i.e. HELSS-treated cells excluded trypan blue dye and did not leak intracellular enzymes into the medium). Collectively, these results demonstrate that HELSS blocks AVP-induced arachidonic acid release by specific and irreversible inhibition of calcium-independent phospholipase A2 and underscore the importance of calcium-independent phospholipase A2 in agonist-induced arachidonic acid release in at least some cell types. PMID- 8407893 TI - Lysophosphatidic acid stimulates mitogen-activated protein kinase activation via a G-protein-coupled pathway requiring p21ras and p74raf-1. AB - Activation of tyrosine kinase receptors causes mitogen-activated protein (MAP) kinase stimulation via a pathway involving p21ras, p74raf-1 (acting as a MAP kinase kinase kinase), and MAP kinase kinases; however, the pathway by which heterotrimeric G-protein-coupled receptors activate MAP kinases is undefined. Since there are several MAP kinase kinase kinases it has been suggested that p74raf-1 may only couple tyrosine kinase receptors to MAP kinase activation. We therefore investigated the requirement for p21ras and p74raf-1 in G-protein receptor-mediated MAP kinase activation. Lysophosphatidic acid stimulates MAP kinase via a pertussis toxin-sensitive pathway, which is blocked by dominant negative Ras. Lysophosphatidic acid-stimulated MAP kinase activation is potentiated by overexpression of p74raf-1 and blocked by expression of a dominant negative Raf protein comprising the N-terminal 259 amino acids. We conclude that lysophosphatidic acid activates MAP kinases by a G-protein-coupled pathway that requires both p21ras and p74raf-1. PMID- 8407894 TI - Interaction between a complex of RNA polymerase III subunits and the 70-kDa component of transcription factor IIIB. AB - A system that detects the formation of complexes between different proteins by linking them to separate domains of the GAL4 transcription activator protein has been used to study protein-protein interactions between four essential and unique subunits of yeast RNA polymerase III (C82, C53, C34 and C31), the 70-kDa component of the initiation transcription factor IIIB (TFIIIB70) and the TATA binding protein. We found that C82, C34, and C31 are able to combine with each other in vivo and that C34 interacts with TFIIIB70. These results suggest that C34 and TFIIIB70 are specificity determinants of the RNA polymerase III-TFIIIB interaction. PMID- 8407895 TI - Follicle-stimulating hormone regulation of A-kinase anchoring proteins in granulosa cells. AB - It has been well established that the biochemical and morphological changes during maturation of granulosa cells that are induced by follicle-stimulating hormone (FSH) occur through the elevation of intracellular cAMP and consequent activation of the cAMP-dependent protein kinase (PKA). In this report we show that FSH action alters the expression of A-Kinase Anchoring Proteins (AKAPs), which function to target the subcellular distribution of the type II PKA. Exposure of granulosa cells grown in primary culture with FSH and estradiol for 72 h resulted in the up-regulation of an 80-kDa AKAP and the RII beta subunit of PKA, whereas cells grown in control medium containing only estradiol produced a time-dependent increase of a 140-kDa AKAP. RII overlays performed with [32P]RII alpha preferentially detected RII-binding bands of 80 and 95 kDa compared to blots probed with [32P]RII beta, suggesting that FSH may alter the subcellular location of PKA in an isoform-specific manner. FSH treatment causes a translocation of RII alpha from the particulate to the cytosolic fraction coincident with the induction of the 80-kDa AKAP, which is also predominately cytosolic. These data suggest that FSH promotes a redistribution of the type II PKA holoenzyme through the selective induction of an RII isoform-specific AKAP. PMID- 8407896 TI - Correction of the class H defect in glycosylphosphatidylinositol anchor biosynthesis in Ltk- cells by a human cDNA clone. AB - Previous attempts to express glycosylphosphatidylinositol-anchored proteins in Ltk- cells have not been successful because Ltk- cells cannot synthesize N acetylglucosamine-phosphatidylinositol, the first intermediate in anchor biosynthesis. Using complementation cloning, we have identified a human cDNA that corrects the defect in anchor biosynthesis and allows the expression of glycosylphosphatidylinositol-anchored proteins in Ltk- cells. The nucleotide sequence predicts a novel cytosolic protein of 188 amino acids. PMID- 8407897 TI - A GTP-dependent step in the activation mechanism of capacitative calcium influx. AB - Calcium influx in electrically non-excitable cells is regulated by the filling state of intracellular calcium stores. Depletion of stores activates plasma membrane channels that are voltage-independent and highly selective for Ca2+ ions. We report here that the activation of plasma membrane Ca2+ currents induced by depletion of Ca2+ stores requires a diffusible cytosolic factor that washes out with time when dialyzing cells in the whole-cell configuration of the patch clamp technique. The activation of calcium release-activated calcium current (ICRAC) by ionomycin- or inositol 1,4,5-trisphosphate-induced store depletion is blocked by guanosine 5'-3-O-(thio)triphosphate (GTP gamma S) and guanyl-5'-yl imidodiphosphate, non-hydrolyzable analogs of GTP, suggesting the involvement of a GTP-binding protein. The inhibition by GTP gamma S occurs at a step prior to the activation of ICRAC and is prevented by the addition of GTP. We conclude that the activation mechanism of depletion-induced Ca2+ influx encompasses a GTP dependent step, possibly involving an as yet unidentified small GTP-binding protein. PMID- 8407898 TI - Three-dimensional structure of rat acid phosphatase in complex with L(+) tartrate. AB - The crystal structure of recombinant rat prostatic acid phosphatase in complex with the inhibitor L(+)-tartrate was determined to 3-A resolution with protein crystallographic methods. The inhibitor binds at the carboxyl end of the parallel strands of the alpha/beta domain. One of the carboxyl groups of the tartrate molecule interacts with the conserved residues Arg-11, His-12, and Arg-15, which form part of the phosphate binding site. Furthermore, the C2 and C3 hydroxyl groups interact with His-257 and Arg-79. The second carboxyl group is close to Arg-79 but makes no direct hydrogen bonds to the protein. A sequence comparison between tartrate-sensitive and -resistant acid phosphatases suggests that these enzymes have different three-dimensional structures. PMID- 8407899 TI - Molecular cloning and functional expression of human acyl-coenzyme A:cholesterol acyltransferase cDNA in mutant Chinese hamster ovary cells. AB - Accumulation of cholesterol esters as cytoplasmic lipid droplets within macrophages and smooth muscle cells is a characteristic feature of early lesions of atherosclerotic plaque. Intracellularly, an essential element in forming cholesterol ester from cholesterol is the enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT). ACAT is a membrane protein located in the endoplasmic reticulum. The ACAT protein has never been purified to homogeneity, and no antibodies directed against ACAT have been reported. The gene(s) encoding this enzyme had not been isolated. This laboratory had previously reported the isolation of Chinese hamster ovary cells expressing human ACAT activity. From DNAs of these cells, we have cloned a 1.2-kb exonic human genomic DNA. This led to the eventual cloning of a 4-kb cDNA clone (K1) from a human macrophage cDNA library. Transfection of K1 in ACAT-deficient mutant Chinese hamster ovary cells complemented the mutant defect and resulted in the expression of human ACAT activity. K1 contained an open reading frame of 1650 bp encoding an integral membrane protein of 550 amino acids. Protein homology analysis showed that the predicted K1 protein shared homologous peptide sequences with other enzymes involved in the catalysis of acyl adenylate formation followed by acyl thioester formation and acyl transfer. These results indicate that K1 encodes a structural gene for ACAT. The cDNA reported here should facilitate future molecular studies on ACAT. PMID- 8407900 TI - Role of the interdomain linker peptide of Trichoderma reesei cellobiohydrolase I in its interaction with crystalline cellulose. AB - Cellobiohydrolase I (CBH I), the major component of Trichoderma reesei cellulolytic system, is comprised of a catalytic core domain joined to a cellulose binding-domain (CBD) by an extended O-glycosylated interdomain linker peptide. Two internal deletions were introduced to the linker in order to investigate its function particularly in the hydrolysis of crystalline cellulose. Deletion of the first one-third of the linker, including a putative hinge region, reduces the binding capacity of CBH I in high enzyme coverage but does not affect its enzymatic activity on crystalline cellulose. The longer deletion removing practically all of the linker dramatically reduces the rate of crystalline cellulose degradation even though the enzyme still binds to the substrate. We conclude that sufficient spatial separation of the two domains is required for efficient function of CBH I. It is evident that the presence of a functional CBD is increasingly important for CBH I toward higher enzyme to cellulose ratios. Our data suggest that the putative hinge removed by the first deletion facilitates CBD-driven binding and dense packing of the wild type enzyme on the cellulose surface. PMID- 8407901 TI - Glutamine 170 to tyrosine substitution in yeast mitochondrial F1 beta-subunit increases catalytic site interaction with GDP and IDP and produces negative cooperativity of GTP and ITP hydrolysis. AB - Glutamine 170 to tyrosine mutation in the beta-subunit from Schizosaccharomyces pombe mitochondrial F1 was found to increase both affinity for ADP, apparent negative cooperativity of ATPase activity, and sensitivity to azide inhibition (Falson, P., Di Pietro, A., Jault, J.-M., Gautheron, D.C., and Boutry, M. (1989) Biochim. Biophys. Acta 975, 119-126). The mutation is shown here to increase the affinity for GDP, IDP, and guanosine 5'-(beta,gamma-imidotriphosphate), which are competitive inhibitors of GTPase and ITPase activities. Various fluorescence approaches also reveal an increased affinity of the catalytic site in mutant as compared with wild-type enzyme for GDP, IDP, and 2'(3')-N-methylanthraniloyl GDP. The mutation alters the maximal rates and pH dependence of GTPase and ITPase activities, whereas wild-type F1 exhibits single optima at pH 7.5-8.0. The pH activity profiles of the mutant enzyme for these substrates are biphasic, with optima at pH 8.5-9.0 and below 6.5. The mutation increases the sensitivity of GTPase and ITPase activities to azide inhibition, which increases with decreasing pH. At pH 6.0-7.0, an apparent negative cooperativity is observed when mutant F1 hydrolyzes GTP or ITP, whereas the wild-type enzyme shows Michaelian kinetics. Addition of bicarbonate at pH 7.0 substantially stimulates GTP or ITP hydrolysis and abolishes the apparent negative cooperativity by the mutant enzyme; on the contrary, the anion produces a slight inhibition of these activities catalyzed by wild-type F1. The overall results suggest that apparent negative cooperativity can be observed with GTP or ITP hydrolysis provided that the release of the respective diphosphate is a rate-limiting step. PMID- 8407902 TI - Two-dimensional, rotational-echo double-resonance NMR of cell culture metabolism. AB - Two-dimensional, rotational-echo double-resonance 13C NMR, a new solid-state NMR technique, has been used to show that the relative fluxes of the labeled chemical bond of L-[2-13C,15N]serine along four metabolic pathways (direct purine synthesis, direct glycine incorporation into protein, direct non-glycyl incorporation into protein, and nitrogen scrambling with loss of carbon) are 1:2:6:36, respectively, for Klebsiella pneumoniae under conditions of nitrogenase derepression. These determinations were performed on a single sample of lyophilized, double-labeled, intact cells. Analysis of the homogeneity of the distribution of label suggests that the primary role of serine in shortening derepression is in providing specific carbon and nitrogen for RNA synthesis. PMID- 8407903 TI - Related signals for endocytosis and basolateral sorting of the asialoglycoprotein receptor. AB - The major subunit of the human asialoglycoprotein receptor contains signals for efficient endocytosis and specific basolateral expression in polarized Madin Darby canine kidney cells, both of which are located within its 40-residue cytoplasmic domain. The aromatic residue in this segment, tyrosine 5, which is necessary for efficient clustering into clathrin-coated pits at the plasma membrane, is also necessary for exclusive basolateral delivery. Mutation of this residue to alanine resulted in a nonpolar expression of the protein. Replacement of tyrosine 5 with phenylalanine yielded almost wild-type rates of endocytosis as well as specific basolateral expression, indicating that tyrosine phosphorylation is not essential for either sorting step. The close similarity between the two sorting signals was further corroborated by deletion mutants showing that the amino-terminal 10 residues of the cytoplasmic domain are sufficient for basolateral polarity and efficient endocytosis. The kinetics of appearance of newly synthesized wild-type and mutant receptor protein at the apical and basolateral surfaces indicate that these proteins are sorted intracellularly and are transported directly to the respective domains. Mutants affected in basolateral sorting lost polarity, i.e, appeared to similar extents on both surfaces, indicating that there is no significant apical sorting information elsewhere in the protein. The close correlation between endocytosis and basolateral polarity suggests common recognition mechanisms at the plasma membrane and in the trans-Golgi network. PMID- 8407904 TI - Asp304 of Escherichia coli acid phosphatase is involved in leaving group protonation. AB - Site-directed mutagenesis was used to explore the role of potential proton donors in the mechanism of the Escherichia coli acid phosphatase that is encoded by the appA gene. Asp304 appeared to be the only carboxylic acid residue that is conserved in the protein sequences of the high molecular weight acid phosphatases. The mutations Asp304Ala and Asp304Glu were introduced into appA and the corresponding proteins were overexpressed in E. coli and purified to homogeneity. Only small decreases were observed for the Km values of the substrates p-nitrophenyl phosphate, fructose 1,6-diphosphate, and tripolyphosphate. However, Vmax was greatly decreased, and the magnitude of effect depended markedly on substrate. Both mutant proteins exhibited significantly lower Vmax values with fructose 1,6-diphosphate, which possesses a much poorer leaving group than do the other two substrates. The importance of the leaving group was further tested by using a number of phenyl and alkyl phosphate derivatives as substrates. A linear correlation was observed between log Vmax and the pKa of the substrate leaving group for catalysis by the Asp304Ala mutant enzyme. These results are consistent with partition experiments using ethylene glycol as an alternate nucleophile, which indicated that for the Asp304Ala protein, the formation of a phosphoenzyme intermediate is the rate-determining step, in contrast to the situation for the wild type enzyme and the His303Ala mutant. In the latter case, the rate-limiting step of the reaction is interpreted to be the breakdown of phosphoenzyme. It is concluded that Asp304, rather than His303, is involved in protonation of the substrate leaving group. PMID- 8407905 TI - Influence of nucleotide binding site occupancy on the thermal stability of the F1 portion of the chloroplast ATP synthase. AB - The irreversible thermal denaturation of the F1 portion of the chloroplast ATP synthase (CF1) was examined by differential scanning calorimetry, ATPase activity loss, and release of bound nucleotides. In nearly all cases, the loss of ATPase activity closely paralleled the temperature dependence of the excess heat capacity. Although the irreversible nature of the denaturation precluded thermodynamic interpretation, a kinetic analysis was feasible. A two-state kinetic model was found to fit the calorimetric data very well. The activation energies of thermal denaturation calculated from calorimetric data were very close to those determined from Arrhenius plots of the apparent first-order rate constants of loss of ATPase activity versus reciprocal of the temperature. The nucleotide binding site occupancy profoundly influenced the temperature at which thermal denaturation occurred. In particular, the temperature at which the maximum in excess heat capacity occurs (Tm) was increased about 8 degrees C by occupancy of tight, noncatalytic ATP binding sites and by an additional 3-4 degrees C by the presence of nucleotides in the medium during heating. The thermal denaturation of CF1, an enzyme composed of nine polypeptide chains, is highly cooperative in that it obeys the simple two-step kinetic model. Since the removal of the epsilon and delta subunits has little effect on thermal denaturation, the major forces that stabilize CF1 must, thus, be between the alpha, beta, and gamma subunits. PMID- 8407906 TI - Plasminogen activation stimulates an increase in intracellular calcium in human synovial fibroblasts. AB - Both plasminogen (Pg) and urinary-type Pg activator (u-PA), but not tissue-type Pg activator (t-PA), bind to normal and rheumatoid arthritis (RA) human synovial fibroblasts in culture with high affinity and in a dose-dependent manner. Single cell intracellular Ca2+ responses to Pg and u-PA were studied using Fura-2 and digital imaging fluorescence microscopy. Pg activation by u-PA on the surface of RA synovial fibroblasts induces a significant rise in cytosolic free Ca2+ concentration ([Ca2+]i) within 90 s. Pg kringle 4 and the alpha 2,3-linked sialic acid in the carbohydrate chain bound to Thr245 are involved in mediating the increases in [Ca2+]i. This response is not observed in normal synovial fibroblasts, suggesting that RA synovial fibroblasts have altered responses to the binding and activation of Pg on their surfaces. PMID- 8407907 TI - Characterization of the third member of the MCAT family of cationic amino acid transporters. Identification of a domain that determines the transport properties of the MCAT proteins. AB - We have identified the third member of a family of cationic amino acid transporters in lipopolysaccharide-stimulated murine macrophages. The deduced amino acid sequence of this transporter is the same as MCAT-2 (mouse cationic amino acid transporter-2), the low affinity transporter expressed in hepatocytes, except for a stretch of 41 amino acids that connects the eighth and ninth membrane-spanning domains. These transporters apparently result from differential splicing of transcripts from a single gene and therefore have been named MCAT-2A (hepatocyte) and MCAT-2B (macrophage). Despite their similarity, MCAT-2B is saturated at one-fifth the arginine concentration, has a lower apparent Vmax, and is more sensitive to trans-stimulation than MCAT-2. Introduction of the unique regions of MCAT-2A and MCAT-2B into the equivalent portion of the related protein, MCAT-1, created chimeric transporters with properties most like the donor of this region. Our findings suggest these 41 amino acids contain a domain that binds the amino acid substrate during its translocation across the membrane. PMID- 8407908 TI - Cloning and characterization of the gene encoding the human platelet glycoprotein V. A member of the leucine-rich glycoprotein family cleaved during thrombin induced platelet activation. AB - Glycoprotein V (GPV) is a major platelet membrane 82-kDa glycoprotein, missing in the Bernard-Soulier syndrome, that is cleaved when platelets are treated with thrombin. We report the cloning and sequencing of the GPV cDNA and gene obtained by a combination of polymerase chain reaction amplification of platelet mRNA and genomic library screening. The single-copy gene for GPV is contained within 6.5 kilobase pairs (kb) of genomic sequence and has a simple structure with a single intron of 958 base pairs in the 5'-untranslated sequence; the coding sequence is contained within a single exon. The promoter region contains a canonical TATA box, and putative GATA, Ets-1, and Sp1 cis-acting elements. Reverse transcription polymerase chain reaction analysis on RNAs from cells of different hematopoietic origins revealed that GPV was specifically transcribed from platelets and from cells of the megakaryocytic lineage (megakaryocytes, HEL cells). A single transcript of 4.5 kb for GPV was detected in human platelets by Northern blot analysis. The entire amino acid sequence of GPV was deduced from the cDNA and genomic sequences. Mature GPV was composed of 544 amino acids which contained a single transmembrane domain, a short cytoplasmic domain (16 residues), and a large extracellular domain with 8 potential N-glycosylation sites. Analysis of the extracellular domain revealed the presence of 15 tandem Leu-rich repeats of 24 amino acids with homology to GPIb alpha and identified a cleavage site for thrombin near the COOH terminus with similarity to the A alpha chain of fibrinogen, but no hirudin-like sequence was found. PMID- 8407909 TI - The single-stranded DNA aptamer-binding site of human thrombin. AB - A new class of thrombin inhibitors based on sequence-specific single-stranded DNA oligonucleotides (thrombin aptamer) has recently been identified. The aptamer binding site on thrombin was examined by a solid-phase plate binding assay and by chemical modification. Binding assay results demonstrated that the thrombin aptamer bound specifically to alpha-thrombin but not to gamma-thrombin and that hirudin competed with aptamer binding, suggesting that thrombin's anion-binding exosite was important for aptamer-thrombin interactions. To identify lysine residues of thrombin that participated in the binding of the thrombin aptamer, thrombin was modified with fluorescein 5'-isothiocyanate in the presence or absence of the thrombin aptamer, reduced, carboxymethylated, and digested with endoproteinase Arg-C. The digestion products were analyzed by reversed-phase high performance liquid chromatography and the peptide maps compared. Four peptides with significantly decreased modification in the presence of the aptamer were identified and subjected to N-terminal sequence analysis. Results indicated that B chain Lys-21 and Lys-65, both located within the anion-binding exosite, are situated within or in close proximity to the aptamer-binding site of human alpha thrombin. The thrombin aptamer binds to the anion-binding exosite and inhibits thrombin's function by competing with exosite binding substrates fibrinogen and the platelet thrombin receptor. PMID- 8407910 TI - The inhibition by fatty acids of receptor-mediated calcium movements in Jurkat T cells is due to increased calcium extrusion. AB - Numerous studies on the molecular basis of the mechanism of action of fatty acids have demonstrated their action in cell signaling and particularly on the regulation of cytosolic Ca2+ concentration. Stimulation of Jurkat T cells with CD3 monoclonal antibody results in an increase of intracellular calcium concentration, [Ca2+]i due both to a release of Ca2+ from intracellular stores and a Ca2+ influx. [Ca2+]i increase represents a dynamic balance between Ca2+ influx and efflux. Fatty acids, either saturated (C14:0), monounsaturated (C16:1), or polyunsaturated, belonging to the C18 and the C20 series induce a marked decrease of CD3-induced [Ca2+]i rise. This property of fatty acid is independent of the position of the carbon-carbon double bond but specific of the cis stereoisomeric form. Fatty acids does not block CD3-induced signals but greatly stimulates the Ca2+ extrusion process probably by activating the plasma membrane Ca(2+)-ATPase. This was documented by the observation that fatty acids, reduced to the same extent as [Ca2+]i, elicited either with CD3 monoclonal antibody the calcium ionophore ionomycin or the Ca(2+)-ATPase inhibitor thapsigargin. PMID- 8407911 TI - A thyrotrope-specific variant of Pit-1 transactivates the thyrotropin beta promoter. AB - Thyrotropin (TSH) beta is a subunit of TSH, the expression of which is limited to the thyrotrope cells of the anterior pituitary gland. Tissue-specific expression of the mouse TSH beta gene is conferred by sequences between -270 and -80 of the 5'-flanking region. We have investigated tissue-specific expression of the TSH beta promoter in two thyrotrope-derived cell types: 1) TtT-97 thyrotropic tumors, which express the endogenous TSH beta gene, and 2) an alpha-TSH cell line, which was generated from a thyrotropic tumor that has lost the ability to express the TSH beta gene. The pituitary-specific transcription factor Pit-1 is present in thyrotropes and interacts with three cis-acting elements in the functionally important region of the TSH beta promoter. Pit-1 protein is present in TtT-97 tumor cells but is absent from alpha-TSH cells. Reintroduction of Pit-1 into alpha-TSH cells by transient transfection does not restore TSH beta promoter activity. We have identified an alternately spliced variant of Pit-1, called Pit 1T, the mRNA and protein expression of which is limited to thyrotrope-derived cells. Pit-1T contains a 14-amino acid insert in the transactivation domain due to an alternate 3' splice acceptor site. Transiently transfected Pit-1T increases TSH beta promoter activity in TtT-97 thyrotropic tumor cells, whereas additional Pit-1 has no effect. The alpha-TSH cell line, which lacks all Pit-1 proteins, requires both isoforms in order to stimulate TSH beta promoter activity. These data suggest that Pit-1T is a thyrotrope-specific splice variant of Pit-1 that is required for TSH beta promoter stimulation; furthermore, both Pit-1 and Pit-1T are required for TSH beta promoter activity in thyrotrope cells. PMID- 8407912 TI - Substrate specificity characterization of a cdc2-like protein kinase purified from bovine brain. AB - A serine/threonine kinase from bovine brain has been purified (Lew, J., Beaudette, K. N., Litwin, C. M. E., and Wang, J. H. (1992) J. Biol. Chem. 267, 13383-13390) and found to consist of a 33-kDa catalytic subunit having high sequence homology to p34cdc2 and cdk2 (Lew, J., Winkfein, R. J., Paudel, H., and Wang, J. H. (1992) J. Biol. Chem. 267, 25922-25926). Substrate specificity determinants for this cdc2-like kinase were examined using synthetic peptide substrates derived from the in vitro p34cdc2 phosphorylation sites of histone H1. The peptide P-K-T-P-K-K-A-K-K-L was found to be an excellent substrate for the bovine cdc2-like kinase, having a Km value in the micromolar range. Important determinants for efficient substrate phosphorylation of this peptide were found both within the proposed substrate consensus motif (S/T-P-X-K/R) of p34cdc2 kinase and outside of this sequence. In addition to the absolute requirement for a proline residue immediately COOH-terminal to the phosphorylatable residue (+1) and a basic residue at the +3 position, a basic amino acid at the +2 position was greatly preferred over an acidic amino acid. A proline residue at the -2 position and a cluster of basic amino acids further COOH-terminal to the consensus motif were also found to be important for substrate binding. HeLa cell p34cdc2 kinase displays similar specificity to that of the bovine cdc2-like kinase, as the additional determinants outside of the consensus motif that contribute to the efficient phosphorylation of the histone peptide by this novel enzyme also appear to be important for p34cdc2-catalyzed phosphorylation. PMID- 8407913 TI - The gamma subunit of the Escherichia coli F1-ATPase can be cross-linked near the glycine-rich loop region of a beta subunit when ADP + Mg2+ occupies catalytic sites but not when ATP + Mg2+ is bound. AB - A mutant of the Escherichia coli F1-ATPase, gamma S8C, has been reacted with a novel bifunctional reagent, N-maleimido-N'-(4-azido-2,3,5,6-tetrafluorobenzamido) cystamine (TFPAM-SS1). Modification of Cys-8 via the maleimide, followed by photolysis to convert the azido group to a reactive nitrene, led to cross-linking of the gamma subunit to a beta subunit. When this cross-linking was conducted with ADP + Mg2+ in catalytic sites, the predominant cross-linked product had a M(r) of 108,000. If cross-linking was done with uncleaved ATP + Mg2+ in catalytic sites, cross-linked products of 102,000 and 84,000 were formed. Cross-linking under both conditions led to inhibition of ATPase activity. TFPAM-SS1 could be cleaved by using reducing agents to break the disulfide bond that links the malemide and tetrafluorophenylazide moieties. Cleavage of this disulfide bond after formation of 102,000 and 84,000 species led to full recovery of ATPase activity. When the 108-kDa cross-linked product was cleaved, full activity was not restored, presumably because of insertion of the tetrafluorophenylazide into a functionally important site on the beta subunit. After cleavage of the disulfide bond, the free thiols could be reacted with [14C]N-ethylmaleimide, thereby radioactively tagging the sites of insertion of the tetrafluorophenylnitrene moiety. In this way, the site of cross-linking from Cys 8 of gamma to the beta subunit in the presence of ADP + Mg2+ was localized to within the sequence Val 145-Lys-155, which contains the glycine-rich loop. This loop region is a part of the catalytic site of the enzyme. PMID- 8407914 TI - The regulation of phospholipase C-gamma 1 by phosphatidic acid. Assessment of kinetic parameters. AB - A survey of lipids revealed that the anionic phospholipid phosphatidic acid activates both control and tyrosine-phosphorylated PLC-gamma 1. The mechanism by which phosphatidic acid activates both forms of PLC-gamma 1 was investigated using kinetic analysis. In the presence of phosphatidic acid, the substrate concentration response for control PLC-gamma 1 changes from sigmoidal to hyperbolic, while the cooperativity index decreases from 2.5 for control to 1.0 for tyrosine-phosphorylated PLC-gamma 1. The primary influence of phosphatidic acid on the control enzyme is on the cooperativity index and not the association of PLC-gamma 1 with substrate micelles, as phosphatidic acid had little effect on the micellar association constant, Ks. Phosphatidic acid also increases the activity of the tyrosine phosphorylated form of the enzyme. This increase is reflected in a decrease in the Km from 0.3- to 0.03-mol fraction phosphatidylinositol 4,5-bisphosphate. Phosphatidic acid has no effect on the Ks of the tyrosine-phosphorylated enzyme. From this data it is concluded that phosphatidic acid appears to activate PLC-gamma 1 by acting as an allosteric modifier. PMID- 8407915 TI - Receptor activation is distinct from hormone binding in intact lutropin choriogonadotropin receptors and Asp397 is important for receptor activation. AB - Hormone binding to receptors on the cell surface triggers a sequence of events (receptor activation and signal generation) leading to activation of effectors in the cytoplasm. Receptor activation and signal generation are difficult to study as both are intimately associated with hormone binding. The lutropin choriogonadotropin (LH/CG) receptor offers a unique model to differentiate and examine receptor activation and signal generation from hormone binding. It belongs to a subfamily of glycoprotein hormone receptors within the G-protein coupled receptor family. This receptor subfamily has several structural features different from the structures of other G-protein-coupled receptors. These receptors consist of a large extracellular N-terminal half and membrane associated C-terminal half of similar size. The truncated N-terminal half alone is capable of high affinity hormone binding, whereas the truncated C-terminal half alone is capable of low affinity hormone binding and cAMP induction. However, this distinction between the high affinity hormone binding and low affinity hormone binding associated with cAMP induction has not been established in intact receptors. As a step to identify a structural element which is responsible for receptor activation and signal generation, we have identified an extracellular Asp of the C-terminal half of the LH/CG receptor which is unique and common to the glycoprotein hormone receptors. Evidence is presented for the first time that Asp397 is important for induction of cAMP synthesis but not essential for hormone binding. Since extracellular Asp397 cannot interact with G protein in the cytoplasm, the inability of the mutant LH/CG receptors with an Asp397 substitution to induce cAMP synthesis is likely to be caused by a defect in the intermediate steps (receptor activation and signal generation) between hormone binding and activation of G-protein. Therefore, our results not only demonstrate that receptor activation and signal generation are distinct from high affinity hormone binding in intact LH/CG receptors, but they also identify an amino acid important for the processes. PMID- 8407916 TI - Folding of maltose-binding protein. Evidence for the identity of the rate determining step in vivo and in vitro. AB - The folding of maltose-binding protein, a periplasmic protein in Escherichia coli, was shown to proceed through the same rate-limiting step whether folding occurred in the cell under physiological conditions or in vitro in the absence of other proteins. Four species of maltose-binding protein containing aminoacyl substitutions identified as decreasing the rate of folding of the protein in vivo were purified, and their denaturant-induced folding transitions were analyzed by monitoring the intrinsic fluorescence of tryptophan. In all four cases the rate of folding in vitro was slower than that of the wild-type maltose-binding protein; thus the same step determines the rate of folding in vivo and in vitro. Furthermore, examination of the three-dimensional structure of maltose-binding protein as determined by x-ray crystallography (F. Quiocho, personal communication; Spurlino, J. C., Lu, G.-Y., and Quiocho, F. A. (1991) J. Biol. Chem. 266, 5202-5219) indicates that all 4 of the residues identified as crucial to folding lie in one structural element of the native protein. We conclude that the rate-limiting step both in vivo and in vitro involves formation of this element of structure. PMID- 8407917 TI - Histidyl-proline diketopiperazine. Novel formation that does not originate from thyrotropin-releasing hormone. AB - We asked whether cyclo(His-Pro) may arise from a substance other than the initially found progenitor thyrotropin-releasing hormone (TRH). Incubation of TRH Gly (pGlu-His-Pro-Gly), a TRH precursor, with brain hypothalamic cytosols caused significant formation of His-Pro-Gly followed by the generation of histidyl proline diketopiperazine (cyclo(His-Pro)), whereas TRH was not formed. The addition of purified pyroglutamyl aminopeptidase resulted in the transformation of TRH-Gly to His-Pro-Gly and cyclo (His-Pro) but not to TRH. Neither TRH nor cyclo(His-Pro) was produced from TRH-Gly in the presence of alpha-amidating enzyme under the present conditions. Nonenzymatic conversion of His-Pro-Gly to cyclo(His-Pro) occurred. The present results provide evidence that cyclo(His-Pro) can emanate from a direct predecessor form of TRH by pyroglutamyl aminopeptidase action, not through TRH formation. PMID- 8407918 TI - Transcriptional activation in yeast by the proline-rich activation domain of human CTF1. AB - Among eukaryotic transcriptional activators, those with acidic activation domains have been shown to stimulate transcription in organisms ranging from yeast to human. In vivo and in vitro assays have been employed to demonstrate transcriptional enhancement in yeast by the proline-rich activation domain of human CTF1. This result suggests that proline-rich activation domains may, like typical acidic activation domains, functional universally. PMID- 8407919 TI - Modulation by vitamin B6 of glucocorticoid receptor-mediated gene expression requires transcription factors in addition to the glucocorticoid receptor. AB - We have investigated the mechanism by which vitamin B6 acts to modulate steroid hormone-mediated gene expression. We show that the level of glucocorticoid induced gene expression from simple promoters, containing only hormone response elements and a TATA sequence, was not affected by alterations in intracellular vitamin B6 concentration. However, modulation of hormone-induced gene expression was restored with the inclusion of a binding site for the transcription factor nuclear factor 1 (NF1) within the hormone-responsive promoter; glucocorticoid induced gene expression was reduced by 44% under conditions of elevated intracellular vitamin B6 concentration and enhanced by 98% in mild vitamin deficiency. Under these conditions, neither glucocorticoid receptor sedimentation characteristics, receptor activation, nor DNA binding capacity was affected. Quantitatively analogous effects were detected with estrogen-induced gene expression when an NF1 binding site was removed from or introduced into an estrogen-responsive promoter. NF1-mediated constitutive transcription was not affected by alterations in vitamin concentration. The modulatory effect of vitamin did not require strict positioning of or spacing between the glucocorticoid response element and NF1 binding site. Moreover, a heterologous transcriptional activator, composed of the viral E1a transactivation domain and the GAL4 DNA binding domain, does not substitute for NF1 in restoring vitamin B6 modulation of hormone-induced gene expression. These results suggest that vitamin B6 modulates steroid hormone-mediated gene expression through its influence on a functional or cooperative interaction between steroid hormone receptors and the transcription factor NF1. PMID- 8407920 TI - Identification of a DNA binding domain in simian virus 40 capsid proteins Vp2 and Vp3. AB - We have identified both biochemically and genetically a protein domain within the simian virus 40 virion protein Vp3, and within Vp2 since its carboxyl two-thirds are identical to the full-length Vp3, that binds DNA in a sequence nonspecific manner. Both the Vp2 and Vp3 (Vp2/3) components of SV40 and mutant SV40(202T) bound either SV40 or pBR322 DNA equally well. Wild type and mutant Vp2/3 proteins, expressed as fusion proteins with glutathione S-transferase (GST), were tested for their ability to bind DNA. GST-Vp3 bound DNA at physiological salt concentrations with an apparent Kd of 2.5 x 10(-8) M and also bound RNA with 4 fold higher affinity. Over 90% of the nucleic acid binding, and all of the activity, was lost upon removal of the carboxyl-terminal 13 and 35 residues, respectively. The DNA binding domain was shown to be distinct and separable from the Vp2/3 nuclear transport signal since mutations within the nuclear transport signal that reduce or abolish nuclear localization of Vp2/3 had no effect on the DNA binding activity of mutant Vp2/3 fusion proteins. The carboxyl-terminal 40 residues of Vp2/3 in the form of a beta-galactosidase fusion protein, F6, are sufficient for DNA binding and may cause compaction of the DNA. The significance of this DNA binding and possible compaction are discussed in relation to the assembly of virion particles. PMID- 8407921 TI - An aspartic proteinase present in seeds cleaves Arabidopsis 2 S albumin precursors in vitro. AB - The Arabidopsis thaliana 2 S albumins are examples of vacuolar proteins which undergo intensive posttranslational processing. An in vitro processing assay to screen for processing enzymes present in seeds was developed using an in vitro synthesized 2 S albumin precursor as the substrate. A protease was characterized which cleaved the substrate into two fragments with molecular weights (as determined from their migration distance on SDS-polyacrylamide gel) corresponding to those of the small and large subunits of Arabidopsis 2 S albumin. The pH optimum of this protease activity, its inhibition by pepstatin A, and partial sequence data led to the conclusion that the protease under study was an aspartic proteinase. Synthetic peptides representing two 2 S albumin propeptides allowed the determination of the in vitro cleavage sites, and suggested that the protease activity is capable, in vitro, of cleaving the amino-terminal propeptide as well as the internal propeptide linking the two subunits. Alterations of the amino acids in and around the cleavage sites, made to study the specificity of the protease activity, suggest that both structural and sequence determinants are important in cleavage site recognition. PMID- 8407922 TI - Cbr, an algal homolog of plant early light-induced proteins, is a putative zeaxanthin binding protein. AB - The cbr gene, previously cloned from the unicellular green alga Dunaliella bardawil, is transcriptionally and translationally activated in parallel to accelerated carotenogenesis in response to light stress conditions. The product of cbr, structurally similar to Elips (early light-induced proteins of higher plants), is associated with a minor light harvesting complexes of photosystem II component (Levy, H., Gokhman, I., and Zamir, A. (1992) J. Biol. Chem. 267, 18831 18836). This study examines the relationship between the induction of Cbr and another plant response to light stress, the deepoxidation of violaxanthin to zeaxanthin. A parallel between the two processes was observed in cells exposed to high light, starved for sulfate, or treated with norflurazon, a herbicide inducing photooxidative damage by inhibiting de novo carotenoid biosynthesis. When highly illuminated cells were returned to normal light, Cbr decayed in parallel to the reepoxidation of zeaxanthin to violaxanthin. Evidence for the physical association of Cbr and zeaxanthin was provided by nondenaturing gel electrophoresis. In cells transferred from low to high light, zeaxanthin was associated with the faster migrating of two electrophoretically resolved fractions of light harvesting complexes of photosystem II that also contained Cbr. In cells growing under normal light, violaxanthin was bound equally to the two fractions. Based on these results we propose that Cbr/early light-induced proteins bind zeaxanthin to form photoprotective complexes within the light harvesting antennae. PMID- 8407923 TI - Metal binding properties of recombinant rat parvalbumin wild-type and F102W mutant. AB - Rat parvalbumin (PV), an EF-hand type Ca(2+)-binding protein, was expressed in Escherichia coli and mutated by replacing a Phe at position 102 with a unique Trp in order to introduce a distinct fluorescent label into the protein. Mass spectroscopy and NMR data indicate that the recombinant wild-type (PVWT) and F102W mutant (PVF102W) proteins have the expected molecular weight and retain the native structure. Both proteins contain two non-cooperative Ca2+/Mg(2+)-binding sites with intrinsic affinity constants, KCa and KMg, of 2.4 +/- 0.9 x 10(7) M-1 and of 2.9 +/- 0.2 x 10(4) M-1, respectively, for PVWT, and KCa and KMg, of 2.7 +/- 1.1 x 10(7) M-1 and of 4.4 +/- 0.3 x 10(4) M-1, respectively, for PVF102W. Based on the highly similar metal binding properties of PVWT and PVF102W the latter protein was used to study cation-dependent conformational changes. Trp fluorescence emission and UV difference spectra of PVF102W indicated that the Trp residue at position 102 is confined to a hydrophobic core and conformationally strongly restricted. Upon Ca2+ or Mg2+ binding the structural organization of the region around the Trp is hardly affected, but there are significant changes in its electrostatic environment. The conformational change upon binding of Ca2+ and Mg2+, as monitored by UV difference spectrophotometry, increases linearly from 0 to 2 cations bound, indicating that the binding of both ions contributes equally to the structural organization in this protein. PMID- 8407924 TI - Dominant negative activity of an endogenous thyroid hormone receptor variant (alpha 2) is due to competition for binding sites on target genes. AB - Regulation of development and metabolism by thyroid hormone (T3) may be influenced by a non-T3 binding T3 receptor (TR) isoform, TR alpha 2, which can inhibit transcriptional activation by legitimate TRs. Numerous mechanisms have been postulated to explain the dominant negative actions of TR alpha 2, including competition for target genes, formation of inactive heterodimers, and squelching. We have found that excess TR alpha 2 was required to inhibit TR alpha 1-mediated transactivation from multiple T3 response elements (TREs). Inhibition of T3 action by TR alpha 2 was specific for TRE-containing genes, because a GAL4/TR alpha 1 chimera, which heterodimerized with the 9-cis-retinoic acid receptor (RXR) and activated transcription from the GAL4 binding site in the presence of T3, was not inhibited by TR alpha 2. In contrast, TR alpha 2 inhibited transactivation by TR alpha/VP16, a chimeric protein containing the N-terminal DNA binding domain (DBD) of TR alpha 1 fused to the transcriptional activation domain of VP16. Indeed, TR alpha 2 inhibited the binding of TR alpha 1 monomers, homodimers, and RXR-heterodimers to DNA in vitro, whereas the TR alpha 2 C terminus alone did not. Although TR alpha 2 bound to TREs with less affinity than TR alpha 1, it bound directly to target genes in the cell nucleus. Furthermore, a TR alpha 2 mutant which binds more avidly to TREs was a more effective inhibitor of T3 action than wild type TR alpha 2. Together these data indicate that TR alpha 2 inhibits T3 action by competing for binding to TREs. PMID- 8407925 TI - GTP binding to elongation factor eEF-2 unmasks a tryptophan residue required for biological activity. AB - Elongation factor eEF-2 from rat liver, which contains 7 tryptophan residues, was treated with increasing concentrations of N-bromosuccinimide (NBS) under conditions in which these residues were oxidized specifically. The reagent produced a characteristic lowering in both the absorbance at 280 nm and the intrinsic fluorescence at 332 nm of the factor. Fluorometric titration of tryptophans and correlation to eEF-2 residual activity on GTP hydrolysis and polyphenylalanine synthesis showed that modification of the two most reactive tryptophans completely inactivated the factor. These residues were identified as Trp343 and Trp221 after cleavage of the protein with cyanogen bromide, separation of the fragments by reversed-phase high-pressure liquid chromatography, and N terminal sequencing of the two fragments which exhibited a decreased absorbance in the NBS-treated protein. Oxidation of the most reactive residue, Trp343, did not induce significant decrease of activity of the factor or of its ability to interact with GTP or GDP. On the contrary, oxidation of Trp221 inactivated the factor, whose residual fluorescence was still partly quenched by GDP but no longer by GTP. Preincubation of eEF-2 with GDP protected Trp221 against NBS oxidation and prevented concomitant inactivation of the factor, whereas preincubation of eEF-2 with GTP increased the sensitivity of the same Trp221 residue to the reagent. Our results show for the first time that Trp221, which is conserved and belongs to a well preserved domain in eukaryotic cells and archaebacteria, plays an essential part in the catalytic activity of eEF-2. They strongly suggest that GTP induces a conformational change of the protein which unmasks this residue, whereas GDP stabilizes a conformation which makes this residue much less accessible. PMID- 8407926 TI - A functional and degenerate pair of EF hands contains the very high affinity calcium-binding site of calbindin-D28K. AB - Calbindin-D28K, a member of the troponin C superfamily of calcium-binding proteins, had six putative EF hand domains containing one very high affinity and two to three lower affinity calcium-binding sites. The location and binding activity of the calcium-binding sites were examined with a recombinant calbindin D28K protein. This protein (Calb I-II) only contained EF hand domains 1 and 2 of calbindin-D28K. Binding of calcium and calcium analogs, the lanthanides, by the recombinant protein was determined in fluorescence emission experiments. Calb I II bound 1 mol of terbium/mol of protein. Terbium was displaced from Calb I-II by other lanthanides and calcium. Fluorescence from terbium was not quenched by holmium. These results and Hill plots of calcium binding activity, determined from intrinsic protein fluorescence measurements, indicated the presence of a single high affinity calcium-binding site on Calb I-II. The properties of the binding site indicated that the very high affinity site of calbindin-D28K was located in EF hand domains 1 and 2 of the protein. In addition, these findings indicated the NH2-terminal pair of EF hands in calbindin-D28K did not depend on interactions with other domains in the protein for high affinity calcium binding activity. The results suggested at least one calcium-binding domain of calbindin D28K can exist as an independent EF hand pair. PMID- 8407927 TI - Isolation and characterization of three Dictyostelium myosin-I isozymes. AB - Using ion exchange chromatography and an ATP-dependent actin precipitation step, we have isolated three myosin-I isozymes that, together, account for most of the K+EDTA-ATPase activity recovered from extracts of Dictyostelium. The two major myosin-I isozymes, present in approximately equal amounts, had apparent molecular masses of 125 kDa on SDS gels and have been identified by amino acid sequence analysis as the products of the Dictyostelium myosin-IB (DMIB) and myosin-ID (DMID) genes. DMIB, with a specific K+EDTA-ATPase activity 10-fold higher than DMID, was responsible for most of the activity in cell extracts. The third isozyme, present in low amounts, had an apparent molecular mass of 137 kDa on SDS gels and is too large to be the product of any of the known myosin-I genes. DMIB eluted from DE53 cellulose columns as two distinct peaks (II and III). Addition of the phosphatase inhibitor okadaic acid to the extraction buffer increased the fraction of DMIB recovered from growth phase cells in peak III from 35 to 70%. DMIB isolated from peak III, but not from peak II, displayed a significant level of actin-activated MgATPase activity. These results indicate that peak III represents a phosphorylated, actin-activatable form of DMIB. PMID- 8407928 TI - Labeling of lysine 492 with pyridoxal 5'-phosphate in the sarcoplasmic reticulum Ca(2+)-ATPase. Lysine 492 residue is located outside the fluorescein 5 isothiocyanate-binding region in or near the ATP binding site. AB - Sarcoplasmic reticulum vesicles were treated with 2 mM pyridoxal 5'-phosphate (PLP) at 25 degrees C and pH 7.0 for 6 min and reduced by NaBH4. Both the activity of the Ca(2+)-ATPase and the capacity for high affinity Mg-ATP binding were greatly reduced. Acetyl phosphate hydrolysis or phosphoenzyme formation from Pi was not inhibited. The enzyme was protected by high affinity Mg-ATP binding against the PLP-induced inhibition. A similar protective effect was obtained by Mg-AMP as well as by Mg-ADP. Acetyl phosphate or Pi gave no protection. The PLP treated vesicles were solubilized in SDS, and the Ca(2+)-ATPase was purified by size exclusion high performance liquid chromatography (HPLC). Mapping the fluorescently labeled peptides in the tryptic digest by reversed phase HPLC revealed a single fluorescent peak, which was protected by Mg-ATP against labeling. Sequencing showed that Lys-492 is the residue labeled with PLP. These results demonstrate that Lys-492 is located in or near the ATP binding site but not in the phosphorylation site or the Pi binding site. When Lys-515 was entirely prelabeled with fluorescein 5-isothiocyanate (FITC), the subsequent labeling of Lys-492 with PLP was not prevented. This finding demonstrates that Lys-492 is located outside the FITC-binding region. It has been widely accepted that FITC occupies the adenosine-binding region within the ATP binding site. In contrast to FITC, Mg-AMP strongly inhibited the labeling of Lys-492 with PLP. These findings lead to the conclusion that Lys-492 is located outside the adenosine-binding region, most probably in or near the region occupied by the alpha-phosphoryl group of Mg-ATP bound to the ATP binding site. PMID- 8407929 TI - Transit and sorting of apolipoprotein B within the endoplasmic reticulum and Golgi compartments of isolated hepatocytes from normal and orotic acid-fed rats. AB - Apolipoprotein B (apoB) secretion by isolated rat hepatocytes was dependent on addition of oleate to the incubation medium and inhibited in hepatocytes isolated from livers of orotic acid-fed rats (OA hepatocytes). To investigate the intracellular transit of newly synthesized apoB under different conditions, normal hepatocytes (with or without oleate) or OA hepatocytes (with oleate) were incubated with [35S]methionine, and subcellular fractions (rough endoplasmic reticulum, smooth endoplasmic reticulum, cis-Golgi, and trans-Golgi and membrane and lumenal contents from these) were isolated at intervals. The specific activities and pool sizes of apoB100 and apoB48 were determined. The observations indicate that there are several points at which intracellular transit of apoB is regulated. Newly synthesized apoB is either translocated to the lumen of the rough endoplasmic reticulum or remains membrane bound and is degraded. The lumenal apoB is either retained and degraded or transferred to the Golgi lumen and secreted. In OA hepatocytes degradation of the membrane-bound form of apoB is inhibited, and the protein accumulates in the trans-Golgi membranes. Although apoB is translocated to the lumen of the rough endoplasmic reticulum in OA hepatocytes, it is not packaged with lipid and is transferred to the Golgi lumen only slowly. PMID- 8407930 TI - Small-angle x-ray scattering studies of the iron-molybdenum cofactor from Azotobacter vinelandii nitrogenase. AB - The nitrogenase enzyme complex, consisting of the molybdenum-iron protein and the iron protein, plays a critical role in the biological reduction of dinitrogen to ammonia (nitrogen fixation). The nitrogen-fixing site within the molybdenum-iron protein is an iron-molybdenum-sulfur cofactor (FeMoco) of roughly 1000-2000 Dalton mass. Structural aspects of FeMoco have been determined by spectroscopic and more recently by crystallographic studies. In order to determine the radius of gyration (Rg) of isolated FeMoco, we have performed small-angle x-ray scattering studies of FeMoco in N-methylformamide solution, in the absence of the molybdenum-iron protein. Model compounds of known structure have also been examined in similar solvents, N,N-dimethylformamide and acetonitrile, as controls and for calibration purposes. The Rg values obtained for the models are in good agreement with calculations based upon their respective crystal structures. However, the Rg obtained for FeMoco clearly indicates that the cofactor is not monomeric in solution, but rather aggregated and possibly polydisperse. Further, Rg values were also measured after addition of thiol, dithionite, and thiol and dithionite, to the FeMoco samples. The results indicate, surprisingly, that oxidation state and putative thiol coordination have no detectable effect on the aggregation behavior of FeMoco in solution, as determined by these measurements. PMID- 8407931 TI - Riboflavin 3'- and 5'-sulfate, two novel flavins accumulating in the roots of iron-deficient sugar beet (Beta vulgaris). AB - Roots from iron-deficient sugar beet grown in the presence of calcium carbonate exhibit a yellow color and autofluorescence typical of flavin-like compounds, whereas roots of control, iron-sufficient plants exhibited no yellow color and extremely low autofluorescence. The two major flavins whose accumulation is induced by iron deficiency have been shown to be different from riboflavin, FMN, and FAD by reversed-phase high performance liquid chromatography. These flavins, accounting for 82 and 15% of the total flavin concentration in deficient roots, have been shown unequivocally to be riboflavin 3'-sulfate and riboflavin 5' sulfate, respectively, by electrospray-mass spectrometry, inductively coupled plasma emission spectroscopy, infrared spectrometry, and 1H nuclear magnetic resonance. These flavin sulfates have not been found previously in biological systems. The localization of riboflavin sulfates in deficient roots is similar, but not identical, to that of high iron reductase activity. The concentration of riboflavin sulfates has been estimated from root extracts to be at least 1 mM. We hypothesize, based on the similar localization of flavin and that of iron reduction, that the accumulation of riboflavin sulfates induced by iron deficiency may be an integral part of the turbo iron-reducing system in sugar beet roots. PMID- 8407932 TI - Calcium plays distinctive structural roles in the N- and C-terminal domains of cardiac troponin C. AB - One- and two-dimensional NMR techniques were used to compare the structural consequences of Ca2+ binding to both the low and high affinity Ca2+ binding sites in recombinant cardiac troponin C (cTnC3). In the absence of Ca2+, the short beta sheet located between the high affinity Ca2+/Mg2+ binding sites in the C-terminal domain was found to be absent or loosely formed as judged by the inter-residue NOEs and chemical shifts of resonances in the Ca2+ binding loops. In contrast, the N-terminal domain beta-sheet located between site II and the naturally inactive site I was present even in the absence of bound Ca2+. Calcium-binding mutant proteins having either an inactive Ca2+ binding site III (CBM-III) or an inactive Ca2+ binding site IV (CBM-IV) (Negele, J. C., Dotson, D., Liu, W., Sweeney, H. L., and Putkey, J. A. (1992) J. Biol. Chem. 267, 825-831) were used to study the structural consequences of Ca2+ binding to each of the high affinity sites located in the C-terminal domain. Only a single active Ca2+ binding site was found necessary for formation of the short beta-sheet between Ca2+ binding sites III and IV. However, the absence of bound Ca2+ at site III was found to produce greater instability in the C-terminal domain as judged from the mobility of the C-terminal aromatic hydrophobic cluster. Thus, Ca2+ binding to the high affinity sites in the C-terminal domain results in an ordering of the aromatic hydrophobic cluster, as well as formation of a short beta-sheet between Ca2+ binding sites III and IV. These results demonstrate that Ca2+ binding plays distinctive structural roles in the N- and C-terminal domains of cTnC. PMID- 8407933 TI - Relationship of low affinity [3]ryanodine binding sites to high affinity sites on the skeletal muscle Ca2+ release channel. AB - Both high and low affinity binding sites for [3H]ryanodine exist in sarcoplasmic reticulum membranes derived from rabbit skeletal muscle. Negatively cooperative binding of [3H]ryanodine at one of four initially identical sites cannot account for some of the kinetic features of the binding to high and low affinity sites. The presence of excess unlabeled ryanodine greatly slows the rate at which [3H]ryanodine bound at the high affinity site dissociates. An examination of the rate of dissociation of [3H]ryanodine bound at increasing [3H]ryanodine concentrations reveals the existence of a second site, occupied only at high ligand concentrations. The occupation of this site correlates well with the conversion of the high affinity site from a site with a dissociation rate constant of approximately 0.0025 min-1 to one with a dissociation rate constant of less than 0.00025 min-1. The low affinity site itself has a dissociation rate constant of 0.013 min-1 and dissociation from this site is unaffected by the presence of 100 microM unlabeled ryanodine. These data suggest that the two binding sites are different but are either allosterically or sterically coupled. Association experiments support this interpretation. Low affinity binding sites for [3H]ryanodine exist in transverse tubule (t-tubule) as well as sarcoplasmic reticulum membranes. High concentrations of both ryanodine and ruthenium red inhibit the binding of [3H]PN200-110 to the dihydropyridine-binding protein in t tubule membranes. Whether the low affinity site in t-tubule membranes is related to that found in sarcoplasmic reticulum membranes is not yet known. PMID- 8407934 TI - Translocation of Rac correlates with NADPH oxidase activation. Evidence for equimolar translocation of oxidase components. AB - Activation of the superoxide-generating NADPH oxidase system of human neutrophils involves the assembly of several neutrophil components, some located on the plasma membrane and others in the cytosol. It has recently been established that one of the required components for NADPH oxidase activity is the GTP-binding protein Rac. To further investigate the role of Rac in the NADPH oxidase system, studies were carried out to determine its subcellular distribution in resting and activated human neutrophils. In resting cells, Rac and an associated guanine nucleotide regulatory factor, GDP dissociation inhibitor (GDI), were located only in the cytosol, along with other known oxidase factors, p47-phox and p67-phox. After activation of neutrophils with phorbol 12-myristate 13-acetate or formyl methionyl-leucyl-phenylalanine, Rac was translocated from the cytosol to the plasma membrane, and this translocation corresponded temporally with the translocation of p47-phox and p67-phox and with the generation of superoxide. GDI remained localized to the cytosol, suggesting activation of the oxidase involved dissociation of the Rac-GDI complex prior to Rac translocation. Determination of the quantities of cytosolic factors associated with the plasma membrane indicated that Rac, p47-phox, and p67-phox are translocated to the plasma membrane simultaneously in equimolar amounts, but that the membrane-associated cytochrome b was present at 3-4-fold molar excess. These findings suggest that Rac may play a role in assembly of the active NADPH oxidase complex. PMID- 8407935 TI - The disulfide folding pathway of hirudin elucidated by stop/go folding experiments. AB - The folding pathway of hirudin was analyzed by structural characterization and stop/go folding experiments of acid-trapped intermediates. The results show that the folding is initiated by a near-random packing, followed by the reorganization and fine adjustment of partially compact intermediates to attain the active molecule. The process of packing is observed as the unfolded hirudin flows sequentially via three groups of equilibrated intermediates, namely one disulfide, two-disulfide, and three-disulfide (scrambled species) isomers. Nearly all possible disulfide species were found to exist along the pathway. Specific tertiary interactions then take effect at the final stage, refining and consolidating the loosely packed intermediates, in the presence of free thiols, to form the active hirudin. The rate of packing and reorganization can be selectively regulated by a number of external factors, and conditions can be chosen to allow the completion of folding process within 10 min or 10 h. PMID- 8407936 TI - Transition state stabilization by chloramphenicol acetyltransferase. Role of a water molecule bound to threonine 174. AB - The structure of the type III variant of chloramphenicol acetyltransferase reveals that Thr-174, a conserved residue, is hydrogen-bonded to a bound water molecule (water 252). Modeling studies (P. C. E. Moody and A. G. W. Leslie, unpublished data) suggested that water 252 could play a part in transition state stabilization via a hydrogen bond to the oxyanion of the putative tetrahedral intermediate. In addition, water 252 is one of three bound water molecules hydrogen-bonded to the 1-hydroxyl group of chloramphenicol in the chloramphenicol acetyltransferase-chloramphenicol binary complex. A combination of site-directed mutagenesis and the use of an alternative substrate has allowed the quantitation of the energetic contribution of each of the interactions made by water 252 to catalysis. Thr-174 was replaced by alanine, valine, and isoleucine, each substitution removing the hydroxyl group hydrogen-bonded to water 252. Steady state kinetic analysis of the mutant enzymes was carried out using both chloramphenicol and 1-deoxy-chloramphenicol as acetyl acceptors. The substitutions at Thr-174 result in a fall in kcat and in decreased affinities for each acetyl acceptor in the binary complexes and also in the ternary complexes with acetyl-CoA. From the calculated free energies in the transition state, the hydrogen bond between water 252 and the oxyanion of the tetrahedral intermediate can be estimated to contribute 0.9 kcal mol-1 toward transition state stabilization, whereas the free energy of the hydrogen bonds between the 1 hydroxyl of chloramphenicol and three bound water molecules provides 1.6 kcal mol 1. PMID- 8407937 TI - A factor binding to the xenobiotic responsive element (XRE) of P-4501A1 gene consists of at least two helix-loop-helix proteins, Ah receptor and Arnt. AB - Xenobiotic responsive element (XRE) is an inducible enhancer element that drives inducible expression of P-4501A1 gene in response to xenobiotic inducers. The XRE binding factor appears in the nuclei of Hepa-1 cells treated with 3 methylcholanthrene (3-MC). Association of the Ah receptor and Arnt (Ah receptor nuclear translocator) in an XRE-binding complex was examined by anti-Ah receptor and Arnt antibodies. Both antibodies inhibited the sequence-specific XRE-binding activity of nuclear extracts from 3-MC-treated Hepa-1 cells and of the cytosolic fraction which was prepared from the nontreated cells and treated in vitro with 3 MC. These results indicate that Ah receptor and Arnt proteins are components of the XRE-binding factor and suggest that Arnt as well as the Ah receptor are localized in the cytosol of nontreated cells. The Ah receptor present in C4 cells, a mutant of Hepa-1 cells defective in the Arnt function, showed an inducer dependent association with Arnt synthesized in an in vitro translation system. Co transfection of the expression plasmids of the Ah receptor and Arnt exhibited synergistically more activated transcription from a reporter gene pMC6.3k consisting of the P-4501A1 gene promoter and enhancer than transfection with either of the two plasmids alone. These findings indicate that the Ah receptor and Arnt proteins form a complex that activates transcription in an inducer dependent manner. PMID- 8407938 TI - Hepatic polysomes that contain apoprotein B mRNA have unusual physical properties. AB - To characterize the association of the mRNA for apoprotein B (apoB) with ribosomes, rat hepatic cytoplasmic extracts were fractionated by density gradient centrifugation. On linear sucrose gradients, the sedimentation velocity of the 14.4-kilobase apoB mRNA was retarded compared to the mRNAs for other hepatic proteins, which were concentrated in fractions containing the bulk of the polysomes. This unusual distribution of apoB mRNA could not be explained by cotranslational association of nascent apoB peptides with lipids, based on experiments using either detergents to delipidate proteins or puromycin to release nascent peptides from polysomes. The results were also not the result of the editing of apoB mRNA, since the sucrose gradient distributions of both edited and nonedited forms were similar. In contrast, the distribution of a 3'-truncated apoB mRNA (apoB-42, 5.8 kilobases) expressed in rat hepatoma cells resembled that of mRNA of a typical hepatic protein. As opposed to the sedimentation velocity results, on equilibrium density gradients most hepatic apoB mRNA was found in the fraction that contained polysomes. Based on these data, the elongation rate of nascent apoB, and the calculated translational yield of apoB mRNA, we conclude that the majority of rat hepatic apoB mRNA must be part of polysomal complexes with unusual physical properties related to the presence of sequence(s) in the 3' region of the message. These sequences may either be primary determinants of structural features or binding sites for protein factors that effect conformational changes. PMID- 8407939 TI - Identification of a skeletal muscle-specific regulatory domain in the rat GLUT4/muscle-fat gene. AB - To identify sequences responsible for the muscle-specific expression of the rat GLUT4/muscle-fat gene, we examined the transcriptional regulation of this gene in the differentiating murine C2C12 skeletal muscle cell line. Differentiated myofibers displayed a 4-5-fold increase in GLUT4 mRNA compared with undifferentiated myoblasts which paralleled the conversion from non-muscle beta actin mRNA to muscle-specific alpha-actin mRNA expression. Transient transfection of progressive 5' and 3' deletions of the GLUT4 5'-flanking DNA identified a 281 base pair region located between -517 and -237 relative to the transcription start site which conferred myotube-specific expression. This region increased reporter activity in the context of the GLUT4 minimal promoter in an orientation independent manner and, in addition, onto the heterologous thymidine kinase promoter. Myotube-specific expression of both GLUT4 reporter constructs and the endogenous mouse GLUT4 mRNA was also observed to be thyroid hormone-dependent. Further, cotransfection of reporter constructs containing the 281-base pair GLUT4 differentiation-specific enhancer with the thyroid hormone receptor specifically increased luciferase activity in myotubes approximately 12-fold. Thus, these data demonstrate the presence of a proximal skeletal muscle-specific activation domain that is necessary for both myotube-specific GLUT4 expression and thyroid hormone responsiveness. PMID- 8407940 TI - Cloning, expression, and chromosomal localization of the mouse meprin beta subunit. AB - Meprins are plasma membrane homo- or hetero-oligomeric metalloendopeptidases that contain glycosylated alpha and/or beta subunits. This paper reports the cloning and sequencing of the mouse kidney beta subunit. The primary translation product is composed of 704 amino acids which includes a transient signal sequence of 20 amino acids at the NH2 terminus. The protease domain (Asn-63 to Leu-260) contains the putative zinc-binding motif characteristic of metalloendopeptidases of the "astacin family." The COOH terminus contains an epidermal growth factor-like domain, a potential membrane-spanning domain, and an additional 26 amino acids. The beta subunit has an overall 42% identity to the alpha subunit, however, a 56 amino acid segment near the COOH terminus of alpha is missing in beta, and the putative transmembrane and cytoplasmic domains of the subunits share no significant sequence similarity. NH2-terminal analyses of detergent-solubilized mature forms revealed that, unlike alpha, the prosequence (Leu-21 to Lys-62) is not removed from the beta subunit. Northern blot analysis revealed a 2.5-kilobase message for the beta subunit in the kidney and intestine of C57BL/6 and C3H/He mice. The gene for the beta subunit was localized to mouse chromosome 18. These studies indicate that alpha and beta probably derived from a common ancestral gene, but have evolved so that their genes are on two different chromosomes, and their tissue-specific expression and post-translational processing differ. PMID- 8407941 TI - A novel CDC2-related protein kinase from Leishmania mexicana, LmmCRK1, is post translationally regulated during the life cycle. AB - The p34CDC2 protein kinase is a key component in the regulation of the eukaryotic cell cycle. We have isolated from the protozoan parasite Leishmania mexicana mexicana a CDC2-related kinase gene (Lmmcrk1) encoding a 34-kDa protein kinase (lmmCRK1) which has 56% amino acid identity with the human CDC2 and contains a PCTAIR motif in place of the highly conserved PSTAIR box. lmmCRK1 was detected in all life cycle stages at comparable levels, yet its histone H1 kinase activity was detected in only the promastigote form, indicating that its activity is stage regulated at a post-translational level. lmmCRK1 did not bind p13suc1 beads and Lmmcrk1 was unable to complement a fission yeast temperature-sensitive cdc2 mutant. These data suggest that Lmmcrk1 is unlikely to be the functional L. mexicana cdc2 homologue. A distinct histone H1 kinase activity that binds p13suc1 beads (SBCRK) was also detected, with activity that correlated with the division status of the developmental forms of the parasite, being present in the dividing stages of the parasite and absent in nondividing metacyclic forms. SBCRK is a candidate for the functional CDC2 homologue, but it does not react with an anti PSTAIR monoclonal antibody on Western blots when eluted from p13suc1 beads, indicating a divergent PSTAIR box. These data suggest that a family of CDC2 related protein kinases are present in Leishmania. Some share sequence and biochemical properties with CDC2, but significant differences also exist, possibly reflecting the evolutionary distance between Leishmania and higher eukaryotes. PMID- 8407942 TI - Characterization of interleukin-10 receptors on human and mouse cells. AB - Human interleukin (IL)-10 is a pleiotropic cytokine acting on a variety of immune cells. Here we show that the protein can be enzymatically iodinated to high specific radioactivity with retention of biological activity. The radiolabeled ligand binds specifically to its receptor in several mouse and human cell lines, notably human B-lymphoma line JY and mouse mast cell line MC/9. Human IL-10 apparently binds as a dimer to a single class of receptor in both the JY and MC/9 cell lines with a Kd in the 50-200 pM range. Interestingly, mouse IL-10 was capable of blocking binding of human IL-10 to mouse but not human cells. There appears to be at most only a few hundred IL-10 receptors/cell for both mouse and human cell lines examined. Chemical cross-linking of the radioiodinated hIL-10 to JY and MC/9 cells revealed a common protein complex with an apparent molecular mass of about 97 kDa. Additional high molecular weight complexes were detected with JY but not MC/9 cells. PMID- 8407943 TI - Interleukin-1-induced signaling in T-cells. Evidence for the involvement of phosphatases PP1 and PP2A in regulating protein kinase C-mediated protein phosphorylation and interleukin-2 synthesis. AB - Binding of human recombinant interleukin-1 beta (IL-1 beta) to the cell surface receptors of EL-4 6.1 murine T-cells results in enhanced phosphorylation of several cellular proteins. Staurosporine abolished the enhanced phosphorylation in response to IL-1 for some of these proteins, suggesting that protein kinase C (PKC) was at least partially responsible. PKC-beta was translocated from the cytosol to the plasma membrane between 2 and 15 min after IL-1 binding. Activation of PKC-beta was enhanced by the protein phosphatase inhibitor okadaic acid. In fact, okadaic acid inhibited dephosphorylation of the PKC-specific peptide GS (Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys-Lys). On the other hand, okadaic acid also led to elevation of IL-1-induced trans/autophosphorylation of PKC-beta. Accordingly, IL-1 induction of interleukin-2 synthesis was markedly enhanced by okadaic acid in EL-4 cells. These data suggest that activation of PKC beta contributes to enhanced phosphorylation of cellular proteins in IL-1-treated cells and support the importance of protein phosphorylation and dephosphorylation in the regulation of IL-1-induced IL-2 synthesis in EL-4 6.1 T-cells. PMID- 8407944 TI - Regulation of epidermal growth factor receptor in NIH3T3/HER14 cells by antireceptor monoclonal antibodies. AB - The mechanism(s) by which monoclonal antibodies (mAbs) against the epidermal growth factor (EGF) receptor regulate receptor function have been investigated with NIH3T3/HER14 fibroblasts expressing human EGF receptors. Bivalent 225 mAb or monovalent 225 Fab' inhibited transforming growth factor (TGF)-alpha-induced EGF receptor tyrosine phosphorylation and cell proliferation. Culture of HER14 cells with 225 mAb or 225 Fab' did not activate EGF receptor tyrosine kinase when assayed after lysis of cells in SDS sample buffer. However, when cells were cultured with bivalent 225 mAb, but not with monovalent 225 Fab', and were subsequently lysed and further incubated in Triton X-100 lysis buffer containing proteinase and phosphatase inhibitors, receptor phosphorylation was observed. Phosphorylation was confined to tyrosine residues and was inhibited by addition of genistein after lysis, indicating that it was due to the activation of protein tyrosine kinase. The activity of bivalent 225 mAb was unphysiologic, in contrast with TGF-alpha, in that receptor kinase activation occurred only after cell lysis and with delayed kinetics; serine and threonine phosphorylation did not occur; and down-regulation of EGF receptors was slower. Selective mAb-mediated phosphorylation of tyrosine residues on EGF receptors was sufficient to activate phosphorylation of a SH2 group-bearing substrate, phospholipase C-gamma, indicating that serine/threonine phosphorylation is not required for EGF receptor kinase activity. These studies provide novel insights into the capacity of bivalent mAb to modulate EGF receptor function. PMID- 8407945 TI - Lignification in cell suspension cultures of Pinus taeda. In situ characterization of a gymnosperm lignin. AB - Pinus taeda suspension cultures grown in medium containing 2,4 dichlorophenoxyacetic acid showed only primary cell wall formation and essentially no lignification, as determined by histochemical, ultrastructural, chemical, and NMR spectroscopic analyses. However, these cultures maintained a functional phenylpropanoid pathway as demonstrated by formation of the lignans ( )-matairesinol and (-)-pinoresinol. Administration of [1-13C]Phe to these cultures, followed by solid-state carbon-13 NMR spectral analysis of their cell walls, demonstrated that the phenylalanine incorporated into the cell wall matrix was primarily as protein, rather than lignin. Successive transfer of the 2,4 dichlorophenoxyacetic acid-grown cultures to alpha-naphthaleneacetic acid containing medium induced cell wall thickening concomitant with lignification. The presence of lignin was confirmed by histochemical, ultrastructural, chemical, biochemical, and NMR spectroscopic analyses. Specific labeling of the lignin polymer in situ with [1-13C]-, [2-13C]-, and [3-13C]Phe and analysis of the cell wall preparations by solid-state carbon-13 NMR spectroscopy permitted the first direct determination of the in situ bonding patterns in a gymnosperm lignin. Several dominant interunit linkages were observed, including beta-O-aryl, furanofuran, phenylcoumarin, and phenolic-linked monolignols, consistent with those predicted but hitherto not proven. Finally, milled wood lignin derivatives prepared from these 13C-specifically enriched lignin tissues gave a relatively high fidelity copy of the native lignin. PMID- 8407946 TI - Regulation by phorbol esters of amyloid precursor protein release from Swiss 3T3 fibroblasts overexpressing protein kinase C alpha. AB - Release of large soluble NH2-terminal fragments of the amyloid precursor protein (APP) of Alzheimer's disease was measured in two Swiss 3T3 fibroblast cell lines (designated SF1.4 and SF3.2), overexpressing the alpha subtype of protein kinase C, and in two control cell lines (SC1 and SC2) (Eldar, H., Zisman, Y., Ullrich, A., and Livneh, E. (1990) J. Biol. Chem. 265, 13290-13296). Basal release of APP was significantly increased in SF1.4 cells, but not in SF3.2 cells, relative to controls. Phorbol 12-myristate 13-acetate, an activator of protein kinase C, elicited a concentration-dependent increase in APP release in all four cell lines. However, the estimated EC50 for this effect was lower in the two cell lines overexpressing protein kinase C-alpha (7 and 6 nM, in SF1.4 and SF3.2 cells, respectively) than in control SC1 and SC2 cells (56 and 22 nM, respectively). The absolute amount of APP released by maximal concentrations of phorbol ester was not altered by overexpression of protein kinase C alpha. The protein kinase C inhibitor H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride) significantly reduced the response to phorbol esters in control (SC1) cells but not in cells (SF1.4) that overexpress protein kinase C alpha. Levels of cell-associated APP were slightly elevated, and rates of APP turnover were unchanged, in SF1.4 cells relative to controls. However, cell-associated APP levels were lower in SF3.2 cells than in controls. The results demonstrate that protein kinase C alpha regulates APP release in Swiss 3T3 fibroblasts, and perhaps in other tissues, including brain, and may be the isozyme that mediates receptor-evoked release of APP. PMID- 8407947 TI - Molecular and cellular characterization of Mex-/methylation-resistant phenotype. Gene and cDNA cloning, serum dependence, and tumor suppression of transfectant strains. AB - Methylating agents are widely distributed environmental mutagens. To identify mechanisms that protect cells against the toxic effects of methylating compounds Chinese hamster ovary (CHO) cells were transfected with human DNA and selected for resistance to methylating agents. From methylation-resistant transfectants, human DNA was cloned that conferred methylation resistance to CHO cells. A human cDNA was isolated (designated c81) encoding a protein of 410 amino acids which is partially identical to human lamp-2 encoding a lysosomal membrane glycoprotein. Transfection of c81, but not of the neo gene or a c81 antisense construct, into CHO cells gave rise to significant increases of survivors after methylating treatments. c81 mRNA is expressed both in human, CHO parental, and transfected cells, albeit in different amounts. The methylation-resistant transfectants lacked alkyltransferase activity (Mex- phenotype), did not repair O6 methylguanine in vivo, and neither showed reduced initial DNA methylation level nor increase of removal of 3-methyladenine and 7-methylguanine from DNA. They are therefore representatives of the "tolerance" phenotype. The transfectants also showed a higher serum requirement, flat growth, and a reduced tumor forming ability. The data suggest a link between altered regulation of cell proliferation and acquired resistance to methylating DNA damaging agents. PMID- 8407948 TI - Complete coding sequence, intron/exon organization, and chromosomal location of the gene for the core I protein of human ubiquinol-cytochrome c reductase. AB - Core I protein is a nuclear-encoded component of the ubiquinol-cytochrome c reductase complex of the mitochondrial respiratory chain. We have located the gene for the human core I protein in the p21 region of chromosome 3, just upstream of the COL7A1 gene which encodes type VII collagen. The core I gene, which has been sequenced in its entirety, is comprised of 10,417 base pairs, from the major transcription start site to the polyadenylation signal, and contains 13 exons. The predicted polypeptide contains 480 amino acids, of which the first 34 are predicted to constitute a typical mitochondrial leader peptide containing 6 positively charged arginine residues. The predicted human protein shows significant homology with core I protein from Saccharomyces cerevisiae, rather high homology (64% similarity, 46% identity) with the processing enhancing protein, which functions as core I protein in Neurospora crassa, and, surprisingly, highest homology with the small subunit of the mitochondrial processing peptidase of rat (74% similarity, 55% identity). The predicted human sequence is 87% identical to the reported bovine core I sequence predicted from cDNA cloning, up to residue 298, but the two predicted sequences are widely divergent after that point. PMID- 8407949 TI - Characterization of two forms of human factor Va with different cofactor activities. AB - Factor Va is an essential cofactor in factor Xa-catalyzed prothrombin activation. Purified human factor Va appears to consist of a heavy chain (M(r) approximately 105,000) and a light chain doublet with M(r) approximately 74,000 and approximately 71,000. We separated factor Va by chromatography on a Mono-S column into two fractions, designated factors Va1 and Va2. Factor Va1 contains the light chain with M(r) approximately 74,000, and factor Va2 exclusively contains the light chain with M(r) approximately 71,000. The two forms of factor Va express different cofactor activities when prothrombin is activated at low phospholipid concentrations or on membranes containing low amounts of phosphatidylserine in phosphatidylcholine. Compared with factor Va2, much higher amounts of factor Va1 are required for factor Xa. Va complex formation at the membrane surface. Once incorporated into the prothrombinase complex, factors Va1 and Va2 are equally active in prothrombin activation. This indicates that the two forms of factor Va do not differ in their ability to promote the catalytic activity of factor Xa or to interact with prothrombin. Direct binding experiments show that the different cofactor activities are explained by a greatly impaired ability of factor Va1 to bind to negatively charged membranes. Factor V is also separated into two protein peaks after chromatography on a Mono-S column. Upon incubation with thrombin, the first peak yields factor Va1 and the second peak factor Va2. The same two forms of factor Va were generated when freshly prepared plasma samples or platelet suspensions were treated with thrombin. This shows that the heterogeneity of the light chain domain is an intrinsic property of both plasma and platelet factor V. It is hypothesized that the heterogeneity is caused by small differences in the carboxylterminal C2 domain of factor V that are introduced as the result of post ribosomal processing. PMID- 8407950 TI - Interaction of high mobility group-I (Y) nonhistone proteins with nucleosome core particles. AB - Mammalian high mobility group (HMG)-I(Y) chromosomal proteins bind with high affinity to the minor groove of A. T-rich sequences of DNA both in vitro and in vivo. Electrophoretic mobility shift assays demonstrate that in vitro both native and recombinant human HMG-I proteins also bind, but with lower affinity, to preferred regions on isolated avian nucleosome core particles containing approximately 146 base pairs of random sequence DNA. Up to four discrete HMG-I core particle complexes can be detected by electrophoretic mobility shift assays when increasing molar ratios of protein are associated with cores. Both protein DNA and protein-protein interactions are involved in HMG-I binding to cores. The interaction of HMG-I with core DNA is demonstrated by both thermal denaturation and DNase I footprinting experiments. Chemical cross-linking studies employing reversible photoactivatable cross-linkers, combined with one- and two-dimensional electrophoretic analyses, indicate that in vitro HMG-I binds to cores in close proximity to histones H2A and H2B and H3. In situ cross-linking of K562 human erythroleukemia cell nuclei demonstrate that native HMG-I(Y) binds in a similar manner to nucleosomal histones in vivo. Proteolytic removal of the positively charged amino-terminal tails of the octamer histones abolishes binding of HMG-I to core particles. However, core binding is not mediated by the negatively charged carboxyl-terminal tail of the HMG-I protein since an in vitro produced mutant protein lacking this region binds to core particles in a manner similar to full-length HMG-I. Together these results demonstrate that HMG-I, both in vitro and in vivo, binds to preferred regions on the front face of core nucleosomes. PMID- 8407951 TI - The carboxyl-terminal transactivation domain of human serum response factor contains DNA-activated protein kinase phosphorylation sites. AB - The serum response factor (SRF) is a 67-kDa phosphoprotein that, together with auxiliary factors, modulates transcription of immediate early genes containing serum response elements in their promoters. Here we show that the carboxyl terminal domain of human SRF is phosphorylated in vivo and is recognized in vitro by the double-stranded DNA-activated serine/threonine-specific protein kinase, DNA-PK. SRF phosphorylation by DNA-PK was stimulated by its cognate binding site. Protein microsequence analysis of a 22-amino acid synthetic SRF peptide and phosphopeptide analysis of genetically altered glutathione S-transferase-SRF fusion proteins identified Ser-435 and Ser-446 of human SRF as sites phosphorylated by DNA-PK. Both serines are followed by glutamine. Changing Gln 436 and Gln-447 to other residues reduced or eliminated phosphorylation by DNA PK, confirming that these glutamines are important determinants for kinase recognition. The carboxyl-terminal transcription activation domain was mapped within a 71-amino acid region that contains both DNA-PK phosphorylation sites. Amino acid substitutions that interfered with phosphorylation by DNA-PK at Ser 435/446 in GAL4-SRF fusion proteins were reduced in transactivation potency. From these data we suggest that DNA-PK phosphorylation may modulate SRF activity in vivo. PMID- 8407952 TI - Yeast MCK1 protein kinase autophosphorylates at tyrosine and serine but phosphorylates exogenous substrates at serine and threonine. AB - The product of the Saccharomyces cerevisiae MCK1 gene is a protein kinase that phosphorylates poly (Glu,Tyr) in vitro and is itself phosphorylated at both tyrosine and serine in vivo. To characterize the substrate specificity of Mck1, the enzyme was purified to apparent homogeneity from the soluble fraction of yeast cell extracts by ammonium sulfate precipitation, followed by ion exchange chromatography (Q- and S-Sepharose), dye-ligand affinity chromatography (Orange A agarose), adsorption chromatography (hydroxylapatite), and ion exchange fast protein liquid chromatography (Mono-S). In the absence of an exogenous substrate, purified Mck1 was able to autophosphorylate on tyrosine and serine. A catalytically inactive mutant (K68R in conserved kinase domain II) expressed in an mck1 delta strain did not contain detectable phosphotyrosine, confirming that the tyrosine phosphorylation observed in vivo is due to autophosphorylation, but did contain phosphoserine, suggesting that Mck1 is a target for other cellular protein kinases. Purified Mck1 phosphorylated a variety of proteins in heat inactivated yeast extracts, primarily on serine (and threonine). The purified enzyme also used a number of mammalian proteins as phosphoacceptors, including myelin basic protein (MBP), microtubule-associated protein 2 (MAP-2), and tau protein. All of these substrates were phosphorylated on either serine or threonine (or both). Mck1 isolated from yeast extracts by immunoprecipitation with an anti-Mck1 antibody directed against its C terminus also phosphorylated MBP at serine. In the same immune complex kinase assay, the K68R mutant did not detectably phosphorylate MBP, indicating that the serine-specific phosphotransferase activity of Mck1 is intrinsic and not due to contamination by an associated kinase. These findings demonstrate that Mck1 is a member of a novel class of protein kinases that displays the ability to phosphorylate all three hydroxyamino acids in proteins. PMID- 8407953 TI - A comparative study of the chymotrypsin-like activity of the rat liver multicatalytic proteinase and the ClpP from Escherichia coli. AB - A comparative study of the chymotrypsin-like activity of the purified recombinant ClpP protease and the multicatalytic proteinase from rat liver is presented. The peptidase activity of both enzymes has been analyzed with several synthetic fluorogenic peptides, containing either aromatic or nonpolar amino acids in their P1 position. The respective Vmax, Km, and Vmax/Km were calculated from kinetic experiments. The substrate specificity of the multicatalytic proteinase, as expressed by Vmax/Km values, indicate the following substrate preference: N-Suc IIW-MCA > N-Suc-LY-MCA > N-Suc-LLVY-MCA > or = N-Suc-AAF-MCA > N-Cbz-GGL-beta-NA > Glut-GGF-beta-NA > FPAM-4-MNA. In the case of the ClpP the order of preference is: N-Suc-LY-MCA > N-Suc-IIW-MCA > N-Suc-LLVY-MCA > or = N-Suc-AAF-MCA > or = N Cbz-GGL-beta-NA > FPAM-4-MNA (where: N-Suc, N-succinyl-; MCA, 7-amido-4-methyl coumarin; beta-NA, beta-naphthylamide; N-Cbz, N-benzyloxycarbonyl-; 4-MNA, 4 methoxy-beta-naphthylamide; Glut, glutaryl. This similar substrate specificity is further supported by the lack of activity of both enzymes against SY-MCA and N Suc-AAPF-MCA (known substrates of chymotrypsin), by very reduced activity against N-Suc-AAA-MCA and by no significant activity against LG-beta-NA. The results of mixed substrate experiments have shown that all the peptides that are substrates seem to be hydrolyzed by a single class of chymotrypsin-like site in both enzymes. The substrate specificity studies suggest a possible evolutionary relationship between the catalytic component of the ClpP of Escherichia coli and the multicatalytic proteinase chymotrypsin-like catalytic component. This conclusion is further supported by other circumstantial evidence: the fact that affinity-purified anti-ClpP antibodies cross-react with two polypeptide components of the rat liver multicatalytic proteinase complex, presented here and also shown previously; the known resemblance of both structures at the electron microscope level; and their reported role in the degradation of NH2-end rule substrates. PMID- 8407954 TI - Ca(2+)-dependent and thapsigargin-inhibited phosphorylation of Na+,K(+)-ATPase catalytic domain following chimeric recombination with Ca(2+)-ATPase. AB - Two chimeric proteins comprising the Na,K-ATPase catalytic domain (large cytosolic loop) and the two flanking regions of the Ca-ATPase were obtained by transient or stable expression in mammalian cells transfected with recombinant DNA. In the first chimera (CpNC), a large portion (containing the nucleotide binding site) of the cytosolic loop between putative membrane spans M4 and M5 of the sarcoendoplasmic reticulum Ca2+ (SERCA) 1 (fast muscle) ATPase was replaced by the corresponding portion of the Na,K-ATPase alpha 1 subunit. In the second chimera (CNpC), an even larger portion (containing the nucleotide-binding site and the phosphorylation site) of the analogous cytosolic loop of the SERCA2 (cardiac muscle) ATPase was replaced by the corresponding portion of the Na,K ATPase alpha 1 subunit. Steady state Ca2+ transport and coupled ATP hydrolysis by the chimeric proteins were negligible as compared to those obtained with SERCA enzymes. Nevertheless, the chimeric proteins were able to utilize ATP to form phosphoenzyme levels equal to those formed by SERCA ATPases. Chimeric and SERCA enzymes exhibited an identical Ca2+ requirement for ATP utilization and sensitivity to thapsigargin (TG) which is a specific inhibitor of SERCA ATPase and not of Na,K-ATPase. Furthermore, both SERCA and chimeric enzymes could be phosphorylated with P(i), and this reaction required removal of Ca2+. In comparative experiments, the functional pattern of seemingly unaffected phosphoenzyme formation and inhibited Ca2+ transport was produced in the SERCA ATPase even by single mutation of Pro337 to Ala, evidently due to defective protein conformation. Retention of Ca2+ and TG sensitivity by the chimeric proteins demonstrates that the Ca(2+)- and TG-binding domains do not reside within the cytosolic loop replaced by chimeric substitution and strongly support previous studies suggesting that binding of calcium required for enzyme activation occurs within the membrane-bound region of the SERCA ATPases (Clarke et al., 1989a; Sumbilla et al., 1991). PMID- 8407955 TI - A growth factor-inducible gene encodes a novel nuclear protein with zinc finger structure. AB - Growth factors rapidly induce numerous genes that encode regulatory proteins, many of which have been identified by cDNA cloning. In this study, differential hybridization was used to screen a cDNA library constructed from human mammary carcinoma MDA-468 cells that were stimulated with transforming growth factor beta 1 (TGF-beta 1) in the presence of cycloheximide. One of the cDNA clones that was induced by TGF-beta 1 was found to have a nucleotide sequence that predicts a 9,450-Da protein with homology to regulatory DNA-binding proteins. This clone was designated metallopan-stimulin-1 (MPS-1) because it encodes a metalloprotein whose mRNA is expressed in a wide variety of actively proliferating cells and tumor tissues. MPS-1 protein contains one "zinc finger" domain of the C4 type, similar to those present in proteins of the steroid/thyroid hormone receptor superfamily and other DNA-binding proteins. The mRNA for MPS-1 was induced in MDA 468 cells by fetal calf serum, TGF-beta 1, TGF-beta 2, and cAMP analogues. The MPS-1 gene is expressed at relatively high levels in several human carcinoma cell lines, particularly those derived from ectodermal layers, and at higher levels in melanomas (ontogenically of neural origin). In contrast, the MPS-1 mRNA is expressed at low levels in normal WI-38 human lung diploid fibroblasts in culture. We hypothesize that MPS-1 protein may play a role as a potentially important mediator of cellular proliferative responses to various growth factors and other environmental signals. PMID- 8407956 TI - Ubiquitin metabolism in cycling Xenopus egg extracts. AB - Xenopus egg extract is capable of supporting mitosis in vitro, which makes it ideal for biochemical analysis of the cell cycle. Since several studies have implicated the ubiquitin system in cell cycle progression, we have measured ubiquitin conjugation rates, proteolysis of ubiquitin-lysozyme conjugates, and rates of isopeptidase activity in cycling Xenopus egg extracts. Although ubiquitin conjugation in cytostatic factor arrested extract was half that in activated extract, there were no changes in rates of ubiquitin conjugation during the cell cycle. Ubiquitin conjugates are degraded by a 26 S ATP-stimulated protease. The ability of the 26 S protease to degrade ubiquitin-lysozyme conjugates and a fluorigenic peptide also remained constant across the cell cycle. In contrast to previously characterized systems, isopeptidase activity in Xenopus egg extract is energy-dependent. Glycerol gradient fractionation of Xenopus egg extract separated two ATP-dependent isopeptidases. On co-sedimented with the 26 S protease; the other sedimented slower and was not associated with any additional proteolytic activity. As found for rates of Ub conjugation and conjugate proteolysis, there was little or no variation in isopeptidase activity during the cell cycle. PMID- 8407957 TI - Hepatocyte growth factor may act as a pulmotrophic factor on lung regeneration after acute lung injury. AB - Hepatocyte growth factor (HGF) has been shown to have hepatotrophic and renotropic functions for regeneration of the liver and kidney through its mitogenic, motogenic, and morphogenic properties. To examine the involvement of HGF in lung regeneration after acute injury, we analyzed changes of HGF mRNA, HGF activity, and HGF receptors in the rat lung after lung insult and measured HGF concentration in sera of patients with various lung diseases. Following the onset of acute lung injury induced by intratracheal hydrochloride injection, a compensatory DNA synthesis occurred in the bronchial epithelium with a peak at 24 h and in the alveolar epithelium with a peak at 48 h. Expression of HGF mRNA in the rat lung remarkably increased only 3 h after the treatment and HGF activity in the lung also increased to about 3-fold at 6 h later. HGF receptors in the lung but not in the other noninjured organs were down-regulated 12 h later. These marked increases in HGF mRNA and HGF activity and the concomitant down-regulation of HGF receptor occurred before the marked compensatory DNA synthesis in bronchial and alveolar epithelial cells. HGF concentration in sera of patients with various lung diseases, as measured by radioimmunoassay, was much higher than that in healthy donors. These results suggest that HGF is newly produced in the lung after acute lung injury and may have a role in regeneration of the lung. PMID- 8407958 TI - The purification of a mismatch-specific thymine-DNA glycosylase from HeLa cells. AB - G/T mispairs that arise in the DNA of higher eukaryotes as a result of spontaneous hydrolytic deamination of 5-methylcytosine to thymine must be corrected to G/C pairs. We describe here the purification to apparent homogeneity of the enzyme that initiates this repair process by excising the mispaired thymine from the hetero-duplex to generate an apyrimidinic site. The enzymatic activity could be attributed to a 55-kDa polypeptide, which was purified from extracts of HeLa cells by a combination of conventional and DNA-affinity chromatography. The enzyme is a mismatch-specific thymine-DNA N-glycosylase, capable of hydrolyzing the carbon-nitrogen bond between the sugar-phosphate backbone of the DNA and a mispaired thymine. In addition to the G/T, the enzyme can remove thymine also from C/T and T/T mispairs in the order G/T >> C/T > T/T. It has no detectable endonucleolytic activity on apyrimidinic sites and does not catalyze the removal of thymine from A/T pairs or from single-stranded DNA. PMID- 8407959 TI - Transactivation by the human T-cell leukemia virus Tax protein is mediated through enhanced binding of activating transcription factor-2 (ATF-2) ATF-2 response and cAMP element-binding protein (CREB). AB - The human T-cell leukemia virus type I (HTLV-I)-encoded transcriptional activator protein Tax is strongly implicated in HTLV-I pathogenesis. Tax regulates HTLV-I gene expression through three 21-base pair (bp) repeat enhancer elements located in the transcriptional control region of the virus. Tax does not bind these elements directly, but mediates transactivation through the cellular transcription factors that recognize a cAMP response element (CRE)-like sequence centered within each of the 21-bp repeats. In this report, we identify activating transcription factor-2 (ATF-2) and CRE-binding protein (CREB) as the principal T cell proteins that bind the three 21-bp repeats in vitro. Purified Tax protein augments the level of RNA synthesis induced by ATF-2 and CREB in a cell-free transcription assay, providing evidence that Tax cooperates with these cellular proteins to activate HTLV-I transcription. Furthermore, Tax dramatically increases the binding of both the T-cell-derived and recombinant forms of ATF-2 and CREB to each of the 21-bp repeats. The target sequences for this enhancement reside within the DNA binding/dimerization domains of these proteins. These data suggest that Tax transactivates HTLV-I gene expression by increasing the number of bound ATF-2 and CREB molecules at the viral promoter. PMID- 8407960 TI - Deletion of a large domain in recombinant human procollagen II does not alter the thermal stability of the triple helix. AB - A construct of the human gene for procollagen II (COL2A1) was prepared with an internal deletion of 5 kilobases that removed 12 exons coding for 291 amino acids from near the NH2 terminus of the triple helix. The construct was then used to transfect stably a human tumor cell line (HT-1080), and clones secreting internally deleted pro alpha 1(II) chain of procollagen II were isolated. The protein was purified, and the thermal stability of the triple-helical domain was assayed by brief protease digestion. The thermal stability of the internally deleted protein was the same as that of intact collagen II even though the triple helix was 39% shorter. Additionally, the thermal stability of the collagenase A fragment was the same as that of the collagenase A fragment of normal collagen II even though it was 38% shorter. Analysis of the results suggested that the thermal stabilities of large fragments of collagen II depended primarily on their contents of -Gly-Pro-Hyp-triplets corrected for length. PMID- 8407961 TI - The interaction of the von Willebrand factor-A1 domain with platelet glycoprotein Ib/IX. The role of glycosylation and disulfide bonding in a monomeric recombinant A1 domain protein. AB - The interaction of von Willebrand factor (vWF) with platelet glycoprotein Ib/IX plays an important role in primary hemostasis. Previous studies have localized the GpIb alpha binding domain of vWF to amino acid residues 449-728, a region containing the vWF-A1 domain. In order to assess the role of A1 domain structure in vWF binding functions, a cDNA encoding residues 475-709 of vWF was expressed in Escherichia coli (non-glycosylated) and in Chinese hamster ovary (CHO) cells (glycosylated). These recombinant proteins contain a single intrachain disulfide bond between C509 and C695 and were purified as monomers with apparent molecular weights of 36,000 (E. coli) and 39,000 (CHO). 35S-Labeled-vWF-A1 proteins bound directly to GpIb/IX receptors on platelets. The non-glycosylated form had a slightly higher affinity (Kd = 1.4 +/- 0.4 microM) than the glycosylated vWF-A1 protein (Kd = 4.5 +/- 0.9 microM) but had similar binding capacity of 28,000 GpIb/IX-specific binding sites per platelet. Additionally, both recombinant vWF A1 proteins bound to heparin but neither bound to immobilized type I and III collagen. Both E. coli- and CHO-derived vWF-A1 proteins inhibited ristocetin induced platelet agglutination with IC50 values of 300 and 700 nM, respectively. Reduction of the only disulfide bond between C509 and C695 abolished platelet binding activity at concentration up to 2 microM of protein. Confirmation of the importance of the 509-695 disulfide bond was obtained from a full-length vWF mutant containing substitutions at C509 and C695 (C509/695S) which failed to bind to the platelet GpIb/IX receptor. These studies document that vWF-A1 domain can bind to GpIb/IX and heparin but not collagen, and that binding to GpIb/IX requires an intact disulfide bond between C509 and C695. Furthermore, glycosylation increases the solubility but reduces binding affinity of recombinant vWF A1. PMID- 8407962 TI - Engagement of specific sites in the plastoquinone niche regulates degradation of the D1 protein in photosystem II. AB - Rapid degradation of the photosystem-II reaction center protein, D1, is driven by visible and ultraviolet irradiance at low photon-flux in the intact plant. We find that all inhibitors of electron flow that replace bound plastoquinone (QB) from its niche on D1 inhibit UV-B-driven D1 degradation, but only some inhibit visible light-driven degradation. Stabilization of the D1 protein by nitrophenol type inhibitors in visible light depends on the dimensions of the side chain at position 6 of the phenyl ring. Likewise, resistance to trypsinization of D1 at Arg-238 and to electron flow inhibitors in D1 mutant V219I (but not A251V, F255Y, S264A, or L275F) are functions of position 6 side chain dimensions in isolated thylakoids. We conclude that the configuration of the QB niche controls D1 protein degradation in intact plants under physiological photon flux. This is true irrespective of the spectral quality of radiation driving degradation. We show that UV-B-driven D1 protein degradation, but not that driven by visible light, requires plastoquinone in the QB niche to proceed. D1 degradation in visible light occurs as long as specific regions at the end of helix D and in the D-de loop of the protein are not engaged. These regions, through substrate (i.e. QB)-mediated stabilization, are proposed to regulate rapid degradation of the D1 protein. PMID- 8407963 TI - RNA binding properties of vaccinia virus capping enzyme. AB - Vaccinia virus capping enzyme, a heterodimer of 95-kDa and 33-kDa subunits, modifies the 5' RNA end and also acts as a transcription termination factor during synthesis of viral early mRNAs. Termination occurs in response to a specific signal, UUUUUNU, in the nascent RNA chain. We now report that purified capping enzyme binds to defined RNAs in solution to form complexes that are stable during native gel electrophoresis. Multiple enzyme molecules can bind to a single RNA. No particular 5' end structure is required for RNA binding, suggesting that the observed protein-RNA interaction is unrelated to the triphosphatase, guanylyltransferase, or methyltransferase functions of capping enzyme. Although binding does not require a UUUUUNU element in the RNA, complex formation is competed preferentially by poly(U) compared to poly(C). Capping enzyme binds to the synthetic 30-mer homopolymers to form a single protein-RNA complex; affinity for U-30 is 10-fold higher than for A-30. The sites of protein RNA contact, as detected by UV cross-linking, are located predominantly within the 95-kDa capping enzyme subunit, which is itself sufficient to bind and cross link to RNA in the absence of the small subunit. PMID- 8407964 TI - The cytoplasmic domain of the H-2Ld class I major histocompatibility complex molecule is differentially accessible to immunological and biochemical probes during transport to the cell surface. AB - An antiserum was generated against a synthetic peptide corresponding to a portion of the cytoplasmic domains of the H-2Ld and H-2Db class I major histocompatibility complex molecules of the mouse. This antibody preparation, R4, binds exclusively to endoglycosidase H-resistant H-2Ld/Db molecules which are not associated with beta 2-microglobulin. Interestingly, acquisition of resistance to endoglycosidase H precedes acquisition of R4 reactivity by 30 min. R4-reactive H 2Ld and H-2Db molecules occur on the cell surface and are phosphorylated in vivo. Other studies show that the tyrosine in the cytoplasmic domain is accessible to radioiodination on only a subset of H-2Ld molecules, and that the two-dimensional electrophoretic profiles of phosphorylated H-2L/Db molecules, of R4-reactive molecules, and of H-2Ld molecules radiolabeled on this cytoplasmic domain tyrosine are virtually identical. R4-reactive H-2Ld molecules do not undergo the peptide- and beta 2-microglobulin-induced conformational changes characteristic of free class I major histocompatibility complex heavy chains. The accessibility of the H-2Ld cytoplasmic domain to R4 and to radioiodination late in biosynthesis and its biological significance are discussed. PMID- 8407965 TI - Inhibition of colony-stimulating factor-1 promoter activity by the product of the Wilms' tumor locus. AB - Colony-stimulating factor-1 (CSF-1) is a member of the immediate early gene family, which is expressed in mitogen-stimulated quiescent fibroblasts. The biological effects of CSF-1 are multifaceted and include stimulation of the proliferation and differentiation of myeloid progenitors and activity of circulating monocytes and tissue-specific macrophages. Ablation of circulating levels of biologically active CSF-1 in mice leads to osteopetrosis and sterility, thus implicating a role for CSF-1 in bone remodeling and implantation. Identification of regulatory elements and cognate transcription factors that bind the csf-1 promoter and mediate such diverse expression patterns is of great interest. We identified a sequence element at -273 to -265 (relative to the transcription initiation site) in the murine csf-1 promoter, which contains overlapping consensus sequences for the Wilms' tumor protein (WT1), EGR-1, SP1, and SP3 proteins. WT1 and EGR-1 proteins produced in vitro bound to this sequence, and co-transfection of wt1 with a csf-1-cat reporter plasmid resulted in repression of promoter activity. Interestingly, nuclear extracts prepared from serum-stimulated C3H10T1/2 cells contained predominantly SP1 and SP3 binding activities, which recognized the -273 to -265 site. Thus repression of the csf-1 promoter by WT1 at this site may involve competition between SP1 family transcriptional activators and the WT1 repressor. Colony-stimulating factor-1 may be a physiologically relevant target gene for regulation by the WT1 transcription factor. PMID- 8407966 TI - Novel isozymes of cAMP-dependent protein kinase exist in human cells due to formation of RI alpha-RI beta heterodimeric complexes. AB - We report that a human neoplastic B cell line (Reh) contains cAMP-dependent protein kinase (cAK) type I (cAKI), but is practically devoid of cAK type II (cAKII). However, these cells contain a novel cAKI isozyme consisting of an RI alpha-RI beta heterodimer in association with phosphotransferase activity (RI alpha RI beta C2) eluting from DEAE-cellulose columns at a salt concentration characteristic of a cAKII. Immunoprecipitation of 8-azido-[32P]cAMP-labeled extracts and DEAE fractions employing specific antibodies directed against RI alpha and RI beta clearly demonstrated the presence of RI alpha-RI beta heterodimers. RI alpha was precipitated with RI beta antiserum and vice versa. Furthermore, disruption of disulfide bridges by reduction-alkylation abolished this coimmunoprecipitation. In addition, formation of heterodimeric complexes of RI alpha and RI beta could be demonstrated in vitro using recombinant RI proteins. Finally, the presence of low levels of RI alpha-RI beta heterodimers could also be demonstrated in human peripheral blood T lymphocytes. RI alpha-RI beta heterodimers complexed with the catalytic subunit represent a novel isozyme of cAKI (RI alpha RI beta C2), which enhances the possibilities for diversification of cAMP-mediated effects. PMID- 8407967 TI - Characterization of a human DNA damage binding protein implicated in xeroderma pigmentosum E. AB - A human DNA damage binding protein implicated in the DNA excision repair disorder xeroderma pigmentosum E was purified to near homogeneity from HeLa cells. The protein is abundant (approximately 10(5) copies/cell) and has a native molecular weight of 154,000-163,000 as estimated by gel filtration and glycerol gradient sedimentation. DNA damage binding activity copurified with polypeptides of 124 and 41 kDa. Based on the native molecular weight, cosedimentation of both polypeptides with DNA damage binding activity on glycerol gradients, and a molar ratio of approximately 1:1 for the two polypeptides, it appears that p124 and p41 are subunits of a heterodimeric protein. Binding to damaged DNA was resistant to K+ concentrations approaching 1 M, but showed anion-specific sensitivity to Cl- concentrations above 0.5 M, suggesting that the majority of the binding energy is contributed by nonionic interactions. In contrast to previous reports, the DNA damage binding protein was shown to recognize cyclobutane pyrimidine dimers in addition to a nonphotoreactivable lesion(s), most likely the pyrimidine pyrimidone (6-4) photoproduct. PMID- 8407968 TI - Comparative analysis of binding of human damaged DNA-binding protein (XPE) and Escherichia coli damage recognition protein (UvrA) to the major ultraviolet photoproducts: T[c,s]T, T[t,s]T, T[6-4]T, and T[Dewar]T. AB - Human cells contain a protein that binds to UV-irradiated DNA with high affinity. This protein, the damaged DNA-binding protein (DDB), is absent from some xeroderma pigmentosum complementation group E cell strains; therefore, it has been suggested that it may be the damage recognition subunit of a human excision nuclease complex. However, the identity of the UV photoproduct bound by DDB and the role of this protein in nucleotide excision repair have been controversial. In this study, we used several synthetic DNA substrates, each of which contains one of the major UV photoproducts, and DDB purified to apparent homogeneity to quantify the specific binding of DDB to various photoproducts. For comparison, the binding of the same photoproducts by the Escherichia coli damage recognition protein UvrA, which is known to be a subunit of the E. coli excision nuclease, was also measured. UvrA and DDB each bound with high affinity to T[t,s]T, T[6 4]T, and T[Dewar]T, but only marginally discriminated between an undamaged oligomer and an oligomer with a T[c,s]T. In contrast to these similarities with regard to the binding to UV photoproducts, UvrA bound to another excision repair substrate, the psoralen-thymine monoadduct, with high specificity, whereas DDB was unable to distinguish between psoralen-adducted DNA and undamaged DNA. We conclude that DDB may play a special role in the repair of UV damage, but it cannot be the sole damage recognition subunit of human excision nuclease. PMID- 8407969 TI - Quantitation of peptide anchor residue contributions to class I major histocompatibility complex molecule binding. AB - Class I major histocompatibility complex molecules play an important role in cellular immunity by presenting antigenic peptides to cytotoxic T cells. Deep polymorphic pockets in the peptide-binding groove of class I major histocompatibility complex molecules provide structural complementarity for peptide "anchor" side chains. However, the minimum requirements of a peptide for high-affinity binding and the contribution of anchor side chains to binding have not been determined yet. To address these issues, we have compared the affinities of various octapeptides for purified, soluble H-2Kb molecules. The results revealed that at least 2 anchor residues are necessary for high-affinity binding, and that high-affinity binding occurs only when anchor side chains are optimally packed within the groove. The estimated free energy contribution of two anchor side chains to binding is unexpectedly large and comparable with that of peptide backbone, suggesting a crucial role of anchor residues in high-affinity, and hence specific, binding to class I molecules. PMID- 8407970 TI - Purification, molecular cloning, and sequencing of phospholipase C-beta 4. AB - We have characterized inositol phospholipid-specific phospholipase C (PLC) isozymes in bovine retina. Chromatography of a retinal homogenate on a heparin column partially resolved six peaks of PLC activity, which differed in their relative selectivities for the substrates phosphatidyl 4,5-bisphosphate (PIP2) and phosphatidylinositol (PI). Five of the peaks were shown to correspond to the known PLC isozymes PLC-beta 1, PLC-beta 3, PLC-gamma 1, PLC-delta 1, and PLC delta 2. PLC-beta 1, PLC-beta 3, PLC-gamma 1, and PLC-delta 1 in the retinal fractions were identified by immunoblotting with isozyme-specific antibodies, and PLC-delta 2 was identified by direct sequencing of tryptic peptides. PLC-gamma 2 and PLC-beta 2 were not detectable by immunoblot analysis. In addition to five of the seven mammalian PLC isozymes identified to date, bovine retina contained a previously unidentified PLC, which exhibited the highest selectivity for PIP2 over PI. The new PLC was purified from a retinal particulate fraction to yield a preparation that contained a major protein band with an apparent molecular mass of 130 kDa on SDS-polyacrylamide gels. Sequence analysis of 12 tryptic peptides derived from the 130-kDa protein suggested that the primary structure of the new PLC is similar to those members of beta-type PLC isozymes, especially to that of PLC-norpA, which was originally identified in Drosophila eye. The new enzyme was thus named PLC-beta 4. A search of a rat brain cDNA library with the polymerase chain reaction and oligonucleotide primers based on common PLC amino acid sequences resulted in the cloning of a rat brain cDNA corresponding to a previously uncharacterized PLC. The cDNA encoded a putative polypeptide of 1176 amino acids, with a calculated molecular mass of 134,532 daltons, that contained the sequences of all 12 tryptic peptides of PLC-beta 4. Furthermore, the deduced amino acid sequence of the encoded protein was more related to PLC-norpA than to any of the three mammalian PLC-beta isozymes. These results suggest that the brain cDNA encodes PLC-beta 4, which is likely a mammalian homolog of PLC-norpA. PMID- 8407971 TI - Function of the hydrophilic carboxyl terminus of type II DNA topoisomerase from Drosophila melanogaster. I. In vitro studies. AB - The function of the hydrophilic carboxyl-terminal region of Drosophila DNA topoisomerase II was examined by constructing a series of deletion mutants at the 3'-end of the Drosophila Top2 cDNA. The truncated enzymes were then expressed in Saccharomyces cerevisiae. Deletion of up to 240 out of 1447 total amino acids had no apparent effect on the enzyme's ability to catalyze topisomerization reactions. When 273, or more, amino acids were deleted, the enzyme was no longer active. Examples were found where deletion of less than 240 amino acids inactivated the enzyme. Based on the hydrodynamic properties determined for one of these mutants, the lack of activity was most likely due to misfolding of the polypeptides. The active mutants have similar hydrodynamic properties and heat inactivation profiles as the intact enzyme, suggesting that they are dimeric and stably folded. The carboxyl-terminal 240 amino acids also were not required for interaction with the drug VM26. The only difference noted between the shortest, active mutant and the full-length enzyme was a decrease in the stability of the interaction of the truncated enzyme with DNA as evidenced by a decrease in the ionic strength at which catalysis was optimal and at which the transition between a processive and distributive mode of supercoil relaxation occurred. PMID- 8407972 TI - Function of the hydrophilic carboxyl terminus of type II DNA topoisomerase from Drosophila melanogaster. II. In vivo studies. AB - Genetic complementation, protein distribution, and in vivo enzymatic activity by carboxyl-terminal truncation mutations of the Drosophila enzyme were examined. Removal of more than 273 of the 1447 amino acids composing the full-length topoisomerase inactivates the enzyme in vivo and in vitro; removal of 227 amino acids or less has no apparent effect on the ability of the enzyme to substitute for a conditional lethal, or null mutation, of the Saccharomyces cerevisiae top2 gene. Four catalytically active mutants, from which 227 or 240 amino acids are deleted, define an intervening, critical region. Each mutant in this critical region displays different in vivo complementation activity ranging from complete complementation to noncomplementation. Deletion analysis revealed a potent nuclear localization signal within the most distal 60 amino acids, although this is apparently not the only functional signal sequence encoded in the enzyme. Subcellular fractionation and indirect immunofluorescence demonstrate that the truncated enzymes localize to the nucleus, albeit with reduced efficiency compared to wild type. The ability of these mutants, including a mutant in the critical region which does not complement, to catalyze the decatenation of replicated plasmids and the segregation of replicated chromosomes was also examined. PMID- 8407973 TI - hsp 70-like protein in rhesus erythrocyte cytosol and its interactions with membrane skeleton under heat and pathologic stress. AB - The rhesus erythrocytes were examined for the presence of protein(s) similar to the 70-kDa class of heat shock proteins (hsp 70). Also, interactions of these proteins with the erythrocyte membrane were studied under heat stress. These cells in their cytosol contained at least two proteins of about 70 kDa molecular mass; one of which closely resembled the hsp 70 family of proteins. This protein under normal conditions localized mainly in the cytosol, but it had a strong tendency to bind the membrane under heat stress. The binding was almost exclusively restricted to the membrane skeleton and seemed to involve primarily the hydrophobic interactions. A 70-kDa protein immunologically similar to the above protein(s) was detected also in the membranes of rhesus erythrocytes harboring the schizont stage of the simian malarial parasite Plasmodium knowlesi. From these results, we conclude that hsp 70-like proteins in the mature mammalian erythrocytes could perhaps play an important role in protecting the cells under stress by stabilizing the membrane skeleton through their interactions with skeletal proteins. PMID- 8407974 TI - Expression and structural analysis of a teleost homolog of a mammalian zona pellucida gene. AB - Gene expression in oviparous vertebrates during vitellogenesis is hormonally regulated. Our laboratory has characterized a unique gene (wf female), which is seasonally expressed in the liver of the female winter flounder Pseudopleuronectes americanus. The wf female mRNA is coexpressed with vitellogenin mRNA and reaches a high level during vitellogenesis. The wf female gene is 2554 base pairs in length and encodes a putative protein of 509 amino acids. The gene consists of eight exons separated by seven introns of different sizes. Within exon 1, there are six PQQ-rich repeats. Four of them encode a putative (PQQ)1PKY polypeptide, similar to the repeats found in the extracellular domains of other proteins. Exons 2-7 share homology with the zona pellucida protein genes rc55 of rabbit and zp-2 of mouse, and the positions of intron boundaries are conserved in the wf female and mouse zp-2 genes. In addition, the transcriptional regulatory cis elements (estrogen response element, CCAAT and TATAAA boxes), as found in vertebrate genes, are also conserved for the wf female promoter and mapped upstream from the initiation site of the wf female primary transcript. PMID- 8407975 TI - Coproporphyrinogen oxidase. Purification, molecular cloning, and induction of mRNA during erythroid differentiation. AB - Coproporphyrinogen oxidase (EC 1.3.3.3), the enzyme involved in the sixth step of heme biosynthesis, was purified to apparent homogeneity from bovine liver; it has a molecular mass of 37,000 daltons. Partial amino acid sequences were determined. Two degenerate oligonucleotides based on the sequences of trypsin-digested peptides were used in a polymerase chain reaction to amplify a 198-base pair fragment of coproporphyrinogen oxidase DNA, using bovine kidney cells cDNA as a starting template. This fragment was used as a hybridization probe to isolate full-length coproporphyrinogen oxidase clones from a mouse erythroleukemia (MEL) cell cDNA library. Sequence analysis revealed that coproporphyrinogen oxidase comprises 354 amino acid residues (M(r) 40,647), with a putative leader sequence of 31 amino acid residues, the result being a mature protein of 323 amino acid residues (M(r) 37,255). RNA blot analysis revealed a 3.0-kilobase coproporphyrinogen oxidase mRNA in mouse liver and in MEL cells. Treatment of MEL cells with dimethyl sulfoxide led to an increase in coproporphyrinogen oxidase mRNA within 10 h, the induction reached a maximum at 24 h, and was in parallel with the induction of ferrochelatase mRNA. The cDNA allows for the expression of active coproporphyrinogen oxidase, the activity of which is mainly present in mitochondria of transfected cultured cells, thereby indicating that mammalian coproporphyrinogen oxidase is mitochondrial enzyme. PMID- 8407976 TI - The epithelial sialomucin, episialin, is sialylated during recycling. AB - Episialin, the sialomucin encoded by the MUC1 gene is abundantly present at the surface of various epithelial cells. It is a transmembrane molecule that matures through several intermediate forms generated by proteolysis and sequential addition and processing of numerous O-linked glycans. Here we demonstrate that one of the biosynthetic intermediates, the premature form, which is incompletely glycosylated appears at the cell surface. By neuraminidase protection assays we show that cell surface-associated forms of episialin are constitutively internalized (0.9% of surface fraction/min) and recycled, with the intracellular residence time and total cycle time being approximately 66 and 143 min, respectively. During recycling, sialic acid residues are added to the premature form of episialin resulting in complete maturation of the molecule. Complete sialylation of episialin requires several rounds of recycling which continues after sialylation is apparently completed. The recycling of episialin may serve to achieve and maintain a high degree of sialylation. PMID- 8407977 TI - Characterization of a new RNA helicase from nuclear extracts of HeLa cells which translocates in the 5' to 3' direction. AB - RNA helicase II, isolated from nuclear extracts of HeLa cells, was purified 1,300 fold and contained a single protein band of 100 kDa when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme displaced partial duplex RNA exclusively in a 5' to 3' direction. This reaction was supported only by ATP and deoxy-ATP at relatively high concentrations (the Km was estimated as 1 mM). The enzyme displayed only ATPase and deoxy-ATPase activity that was stimulated preferentially by poly(C). RNA helicase catalyzed the unwinding of duplex RNA and RNA.DNA hybrids provided that single-stranded (ss) RNA was available for the helicase to bind. In the presence of MgCl2 and ATP or adenosine 5'-O-(3-thiotriphosphate) RNA helicase II interacted with ssRNA and yielded a protein-RNA complex only when reaction mixtures were treated with glutaraldehyde after incubation. When the reactions contained nonhydrolyzable ATP analogs or GTP, ssRNA was converted into an electrophoretically slower migrating form by the helicase. The slower migrating RNA form was shown to be an RNA species containing secondary structure that resided within a putative hairpin loop. These observations indicate that RNA helicase II can introduce intramolecular secondary structure in ssRNA. PMID- 8407978 TI - Horseshoe crab coagulation factor B. A unique serine protease zymogen activated by cleavage of an Ile-Ile bond. AB - Horseshoe crab factor B is an intracellular serine protease zymogen involved in the bacterial endotoxin-responsive hemolymph coagulation cascade. cDNAs for factor B were isolated utilizing a polymerase chain reaction product using two primers derived from the partial amino acid sequence. The cloned cDNA of 1928 base pairs encoded 400 amino acid residues of factor B precursor. The first 23 amino acid residues constitute a presumed prepropeptide that may be processed by both a signal peptidase and a processing protease, similar to mammalian vitamin K dependent protease precursors. The mature protein consists of 377 amino acids with a calculated molecular mass of 40,570 Da. The overall structure is highly homologous to that of limulus proclotting enzyme (35.9% identity), the substrate for active factor B in the cascade. Like the proclotting enzyme, mature factor B is composed of an amino-terminal "clip"-like domain and a carboxyl-terminal serine protease domain homologous to that of human plasma prekallikrein (36.5%). Internal sequences encode a unique activation peptide. Surprisingly, the cleavage sites of the zymogen factor B for activation by limulus active factor C were found to be an Arg-Ser and an Ile-Ile bond, the latter of which has not been found in any other protease zymogens. These cleavages result in the release of the activation peptide, which consists of 21 residues with a carboxyl-terminal isoleucine. These results indicate that the intracellular clotting system of the limulus hemocyte, like mammalian plasma clotting cascade, proceeds with the sequential activation of three serine protease zymogens: factor C, factor B, and proclotting enzyme. PMID- 8407979 TI - Position-specific Asp-Lys pairing can affect signal sequence function and membrane protein topology. AB - Positively charged amino acids are major determinants of the topology of bacterial inner membrane proteins, whereas negatively charged residues by themselves have little or no influence on the transmembrane orientation. Further, positively charged amino acids can very efficiently block the function of signal sequences when placed immediately downstream, while negatively charged residues are much less potent also in this regard. Here, we show that a negatively charged aspartic acid situated close to a positively charged lysine can attenuate both of these effects in a position-specific manner, suggesting that intra- or intermolecular charge pairing can modulate the interactions between positively charged residues in the nascent chain and parts of the secretory machinery or membrane phospholipids. These observations further underscore the importance of charged amino acids during protein translocation and membrane protein assembly. PMID- 8407980 TI - Purification and characterization of endothelin-converting enzyme from rat lung. AB - Endothelin-converting enzyme, a key enzyme in the production of a potent vasoconstricting peptide, endothelin, was purified to homogeneity from rat lung microsomes. A sensitive and convenient assay method using the 125I-endothelin-1 receptor binding assay was developed for purification studies. The enzyme was solubilized with Triton X-100 and was purified at high yield by sequential column chromatography on wheat germ agglutinin-agarose, zinc-chelating Sepharose, and Blue Bagarose. Electrophoretic analysis of the purified enzyme revealed one protein band with M(r) 130,000. High performance liquid chromatographic analysis of the enzyme reaction revealed that purified enzyme quantitatively converted big endothelin-1 to endothelin-1, indicating that the enzyme specifically cleaved the bond between Trp21 and Val22. The enzyme was inhibited by metal chelators and phosphoramidon, but not by thiorphan and captopril. Lung endothelin-converting enzyme preferred big endothelin-1 to big endothelin-2 or -3 as a substrate, and kinetic analysis revealed that the Michaelis constant and a maximal velocity for big endothelin-1 were 0.20 microM and 3.1 nmol/min.mg protein, respectively. Lung endothelin-converting enzyme is a typical neutral metalloproteinase and is similar to the enzyme from endothelial cells. PMID- 8407981 TI - Core glycosylation of cytochrome P-450(arom). Evidence for localization of N terminus of microsomal cytochrome P-450 in the lumen. AB - It was found that cytochrome P-450(arom) purified from human placenta microsomes is glycosylated, and the sugar chain was cleaved with endoglycosidase H (Endo H). The core glycosylation of P-450(arom) was examined with two heterologous expression systems, cultured insect cells and in vitro translation system. The P 450(arom) protein expressed in the insect cells was glycosylated, and the sugar chain was sensitive to Endo H. It was also glycosylated when translated with the wheat germ cell-free system in the presence of rough microsomal membrane, and the sugar chain could be removed by Endo H treatment. Since the P-450(arom) molecule has two potential glycosylation sites (Asn-12 and Asn-180), we replaced each of the 2 asparagine residues with alanine by site-directed mutagenesis and examined the glycosylation of the two mutant proteins in the cell-free system. The core glycosylation did not occur when the Asn-12 residue was mutated, whereas the mutant protein with modified Asn-180 residue was glycosylated. These results demonstrated that the potential glycosylation site (Asn-12) in the N-terminal portion of P-450(arom) is the site of glycosylation. We conclude that the N terminus of P-450(arom) is translocated across the endoplasmic reticulum membrane to be glycosylated at the luminal side. PMID- 8407982 TI - Apolipoprotein A-I domains involved in lecithin-cholesterol acyltransferase activation. Structure:function relationships. AB - A series of mutant apolipoprotein (apoA-I) constructs were designed and then expressed in cell culture to identify structural domains within the mature native apoA-I protein that participate in the activation of the plasma enzyme, lecithin cholesterol acyltransferase (LCAT). Evolutionary conservation analysis has shown previously that apoA-I contains eight repeats containing 22 amino acids and two repeats containing 11 amino acids that are highly conserved among species as well as within the apolipoprotein supergene family. These tandem repeats begin at residue 44 and are usually marked by a proline residue, with six of the 22-mer repeats showing high amphipathic alpha-helical character. To determine if specific 11- or 22-amino acid domains are essential for maximal LCAT activation within the entire native protein, each of the 10 repeats was sequentially deleted using a polymerase chain reaction based method of mutagenesis. The wild-type and mutant apoA-I gene constructs were expressed in Chinese hamster ovary (CHO) cells and stable lines established. Wild-type and mutant apoA-I protein were purified from 48-96-h conditioned serum-free medium and characterized by SDS polyacrylamide gel electrophoresis and Western blot analysis. Wild-type apoA-I showed a single migrating band of 28,000 daltons that corresponded to the mobility of human plasma apoA-I, whereas apoA-I deletion mutants (lacking 22- or 11-mer repeats) showed the corresponding shift to lower molecular size. To measure the relative LCAT activation of all deletion mutant apoA-I proteins relative to wild-type apoA-I, an assay system utilizing small unilamellar vesicles as the lipid substrate was used. The results of these studies suggest that several central amphipathic alpha-helical regions within the mature protein are critical in LCAT activation. PMID- 8407983 TI - Association of protein kinases ERK1 and ERK2 with p75 nerve growth factor receptors. AB - Extracellular signal-regulated protein kinases (ERKs) constitute a family of protein serine-threonine kinases implicated in a variety of cell-signaling pathways. In cultured rat pheochromocytoma PC12 cells, ERK1 and ERK2 are activated by nerve growth factor (NGF), which also induces rapid association between ERK1 and the high affinity gp140prototrk tyrosine kinase NGF receptor. In the present work, we investigated the possible association between ERKs and the low affinity NGF receptor, p75. Extracts of PC12 cells (before and after NGF treatment) were subjected to immunoprecipitation with anti-p75 antibodies or antiserum; the immune complexes were then assessed for the presence of ERK proteins and tyrosine phosphorylation or for ERK activity using a specific substrate peptide. ERK1 and, to a lesser extent, ERK2 were found to be constitutively associated with p75. NGF did not modulate the total amount of ERK proteins coimmunoprecipitated with p75 but did markedly stimulate the level of p75-associated ERK catalytic activity. NGF treatment also enhanced the tyrosine phosphorylation of a p75-associated species that co-migrates with ERK1 in Western blots. Finally, K-252a, a compound that specifically inhibits activation by NGF of gp140prototrk, abolished the latter effect. These findings indicate that NGF, via activation of gp140prototrk, leads to association of enzymatically active ERKs with p75 and raise the possibility that this interaction may play a role in the NGF mechanism of action. PMID- 8407984 TI - Phosphatidylserine decarboxylase from Saccharomyces cerevisiae. Isolation of mutants, cloning of the gene, and creation of a null allele. AB - Phosphatidylserine decarboxylase plays a pivotal role in the synthesis of phospholipid by the mitochondria. The substrate phosphatidylserine is synthesized extramitochondrially and must be translocated to the mitochondria prior to decarboxylation. To understand the properties of the decarboxylase and exploit its unique topology to address basic questions of interorganelle cooperation in membrane assembly, we have begun to examine this enzyme in Saccharomyces cerevisiae. Strains of the yeast defective in enzyme activity were isolated by modified brute force using 1-acyl-2[N-(6-[7-nitrobenz-2- oxa-1,3-diazo-4 yl)]aminocaproyl] (NBD)-phosphatidyl[1'-14C]serine as substrate for permeabilized cells. Mutant strains with less than 5% wild type activity exhibited no defective growth phenotype. The gene for the phosphatidylserine decarboxylase (PSD) was cloned using an oligonucleotide probe degenerate for the sequence VGAT(I)/(N)VGSI, which is the longest stretch of sequence identity between the Escherichia coli sequence (I at position 5) and the putative CHO cell sequence (N at position 5). The gene encodes a 500 amino acid protein with 28-43% identity to the bacterial and mammalian sequences. The yeast PSD gene maps to the long arm of chromosome 14 between the kex 2 and RAS 2 loci. Null mutations created by disrupting the PSD gene with TRP1 demonstrate that the gene is not essential for cell growth even when the engineered strains are deprived of choline and ethanolamine. Analysis of lipid synthesis and enzyme activity in null mutants indicates that there are two PSD genes. PMID- 8407985 TI - Structure and function of a membrane anchor-less form of the hemagglutinin neuraminidase glycoprotein of Newcastle disease virus. AB - The hemagglutinin-neuraminidase (HN) glycoprotein of paramyxoviruses is anchored in the virion membrane near its amino terminus, protruding from the virion surface to mediate attachment to cellular receptors. Solubilization of HN spikes can be achieved by treatment of virions with detergent and high salt concentrations. When the solubilized HN protein from the Australia-Victoria (AV) isolate of the virus is incubated at 37 degrees C, a chymotrypsin-sensitive site between residues 112 and 113 is exposed. A chymotrypsin-cleaved soluble form of the protein, named CT-HN, has been prepared using this approach. It is membrane anchor-less, due to removal of a 14-kDa fragment from the NH2 terminus of HN. It retains all potential glycosylation sites and cysteines present in the ectodomain of the native protein. It migrates in nonreducing gels and sediments in sucrose gradients at the rate expected for homodimeric HN. The latter is also consistent with our demonstration by site-directed mutagenesis that cysteine residues at positions 6 and 123, respectively, mediate disulfide-linked homotetramer and homodimer formation. CT-HN retains almost total antigenicity, suggesting that it is conformationally very similar to the intact molecule, as well as receptor recognition function and, at low pH, neuraminidase activity. It should prove to be a useful tool for further studies of the structure and function of this important viral glycoprotein. PMID- 8407986 TI - Characterization of the receptor responsible for thrombin-induced intracellular calcium responses in osteoblast-like cells. AB - The receptor responsible for the increase in intracellular calcium concentration ([Ca2+]i) after the addition of thrombin to the human osteoblast-like cell line Saos-2 has been characterized. Thrombin caused a dose-dependent increase in [Ca2+]i; a half-maximal stimulation was observed with 3.2 +/- 1.1 nM thrombin. The human platelet thrombin receptor is activated by thrombin cleavage to create a new NH2 terminus that acts as a tethered ligand, and peptides based on the tethered ligand can activate the receptor independently of thrombin. Northern analysis indicated the presence of mRNA encoding the platelet receptor in Saos-2 cells, and surface expression of the receptor was demonstrated by immunocytochemistry. A tethered ligand peptide (SFLLRNPNDKYEPF, single-letter amino acid code) was found to increase [Ca2+]i. The maximal response to the peptide was similar to that observed with thrombin, and a half-maximal response was observed with 22 +/- 6 microM peptide. The time course of the increase in [Ca2+]i with the peptide was different than that observed with thrombin; a pronounced shoulder was observed after an initial sharp rise. The phenylalanine in the second position of the agonist peptide and the arginine in the fifth position were shown to be essential for its activity. The requirement for proteolysis of the receptor for the thrombin-dependent increase in [Ca2+]i was demonstrated by two methods. Antibodies that reacted with the cleavage site of the receptor abolished the effect of thrombin on [Ca2+]i. In addition, a mutant of thrombin without catalytic activity as well as chemically inactivated thrombin failed to cause an increase in [Ca2+]i. Similar results were obtained with the rat osteoblast-like cell line UMR-106; a tethered ligand peptide based on the rat sequence induced an increase in [Ca2+]i, and antibodies to the cleavage site of the rat receptor inhibited the effect of thrombin. PMID- 8407987 TI - Effects of 5-fluorouracil substitution on the RNA conformation and in vitro translation of thymidylate synthase messenger RNA. AB - In vitro transcribed thymidylate synthase (TS) mRNA which is 100% substituted with 5-fluorouracil (FUra) was analyzed for changes in mRNA secondary structure, for alterations in translational efficiency, and for evidence of translational miscoding in vitro. FUra substitution in TS mRNA results in an altered migration pattern in non-denaturing RNA gels and in decreased hyperchromicity in RNA melting temperature studies, consistent with a change in mRNA secondary structure. However, no change in the translational efficiency of FUra-substituted TS mRNA is seen compared to control TS mRNA in either rabbit reticulocyte lysate or wheat germ extract in vitro translation systems. Analysis of the in vitro translation product of FUra-substituted TS mRNA by Western immunoblotting, isoelectric focusing, 5-fluoro-2'-deoxyuridine 5'-monophosphate binding, and TS catalytic activity experiments shows no difference compared to control TS mRNA. We conclude that the in vitro translation products of FUra-substituted and control TS mRNA are identical. Our findings do not support the hypothesis that changes in the mRNA template are responsible for the RNA-directed cytotoxicity of FUra. PMID- 8407988 TI - Enhanced phosphorylation and dephosphorylation of a histone-like protein in response to hyperosmotic and hypoosmotic conditions. AB - Placement of endothelial cells under hypoosmotic or hyperosmotic conditions results in the reduction and increase, respectively, in the phosphorylation of a M(r) = 16,500 protein (P17). The changes were dose-dependent with a 3.3 +/- 0.3 fold increase occurring at 485 mosm/kg H2O and negligible phosphorylation observed at 202 mosm/kg H2O. The phosphorylation and dephosphorylation were rapid and prolonged; modified phosphorylation levels were maintained as long as the anisotonic conditions were present. However, return to isotonic medium reversed the phosphorylation back to normal within 1 h. Cellular fractionation studies showed that P17 was associated only with the nuclear compartment under isotonic, hypertonic, or hypotonic conditions. Two forms of P17 with pI values of 9.2 and 9.6 were resolved by isoelectric focusing; both forms showed enhanced phosphorylation by hyperosmotic treatment. Phosphorylation occurred on serines exclusively. These studies demonstrate that a nuclear protein with characteristics similar to histones is affected by cell shrinkage or swelling through changes in its phosphorylation state. PMID- 8407989 TI - Release from G1 growth arrest by transforming growth factor beta 1 requires cellular ras activity. AB - Transforming growth factor beta 1 (TGF beta 1) is a potent inhibitor of epithelial cell growth, although the mechanism of growth inhibition remains unknown. We report here a critical relationship between cellular p21ras activity and TGF beta 1 action. Microinjection of oncogenic Ha-ras protein into TGF beta 1 arrested mink lung epithelial cells overcomes TGF beta 1 growth inhibition and allows progression into S phase. Cells released from TGF beta 1 inhibition following microinjection with anti-p21ras antibody, on the other hand, remain TGF beta 1-arrested and do not enter S phase, indicating a requirement for p21ras activity. These biological data are substantiated biochemically in that TGF beta 1 is shown to decrease the activation state of endogenous p21ras, as measured by the level of GTP-bound p21ras. In addition, the phosphorylation and kinase activity of mitogen-activated protein kinase, which depends upon cellular ras activity, is elevated in cells which have been released from growth arrest by TGF beta 1. Together these data demonstrate the involvement of p21ras activity in TGF beta 1-induced growth inhibition and suggest that the inhibitor controls proliferation by modulating the activity of p21ras. PMID- 8407990 TI - Identification of a novel serum and growth factor-inducible gene in vascular smooth muscle cells. PMID- 8407991 TI - Regulation of human NAD(P)H:quinone oxidoreductase gene. Role of AP1 binding site contained within human antioxidant response element. PMID- 8407992 TI - The role of heat shock proteins in regulating the function, folding, and trafficking of the glucocorticoid receptor. PMID- 8407993 TI - A bacterial enzyme that catalyzes formation of carbon monoxide. AB - We have isolated and purified an enzyme (E-2) from Klebsiella pneumoniae, which catalyzes the formation of CO from CH3-S-CH2-CH2-CO-C(OH) = CH-O- (III). This compound is an intermediate in the conversion of 5'-methylthioadenosine to methionine. Concomitant with CO formation, methylthiopropionic acid and formate are produced and O2 is consumed. E-2 also catalyzes the formation of CO, formate, and butyrate from CH3-CH2-CH2-CO-C(OH) = CH-O- (IIIa), the desthio analog of III. Experiments with isotopic IIIa have shown that formate is derived from 1-C, and CO from 2-C. E-2 has a M(r) = 18,500 and requires Mg2+, and no chromophoric cofactor has been detected. PMID- 8407994 TI - Phosphorylation of the cytoplasmic tail of the 300-kDa mannose 6-phosphate receptor is required for the interaction with a cytosolic protein. AB - The cytoplasmic tail of the human 300-kDa mannose 6-phosphate receptor (MPR 300 CT) is an excellent substrate for casein kinase II in vitro. The phosphorylated MPR 300-CT was cross-linked by means of bis(sulfosuccinimidyl)suberate mainly to a cytosolic protein of 35 kDa (referred to as TIP 35) and to 35- and 91-kDa proteins salt-washed from bovine brain membranes. Gel filtration suggested that TIP 35 is part of a higher molecular mass complex of approximately 130-150 kDa. Inhibition studies, using non-phosphorylated and phosphorylated MPR 300-CT and cross-linking, indicate that the interaction with TIP 35 is phosphorylation specific. Furthermore, TIP 35 was only cross-linked to the MPR 300-CT phosphorylated by casein kinase II whereas the MPR 300-CT phosphorylated by protein kinase A failed to cross-link to TIP 35. These results indicate that the cytoplasmic tail of the MPR 300 interacts with a cytosolic protein depending on the phosphorylation by a casein kinase II-like kinase. The cross-linking with salt-washed membrane proteins, however, is inhibited by non-phosphorylated MPR 300-CT, suggesting that different structural determinants in the MPR 300-CT interact with cytosol- and membrane-derived proteins. PMID- 8407995 TI - Isolation of the gene for murine glucose-6-phosphatase, the enzyme deficient in glycogen storage disease type 1A. AB - Glycogen storage disease (GSD) type 1a (von Gierke disease) is caused by a deficiency in glucose-6-phosphatase, the key enzyme in glucose homeostasis catalyzing the terminal step in gluconeogenesis and glycogenolysis. Despite its clinical importance, this membrane-bound enzyme has eluded molecular characterization. Here we report the cloning and characterization of a murine glucose-6-phosphatase cDNA by screening a mouse liver cDNA library differentially with mRNA populations representing the normal and the albino deletion mouse known to express markedly reduced glucose-6-phosphatase activity. Additionally, we identified the gene that consists of 5 exons. Biochemical analyses indicate that the in vitro expressed enzyme is indistinguishable from mouse liver microsomal glucose-6-phosphatase exhibiting essentially identical kinetic constants, latency, thermal lability, and vanadate sensitivity. The characterization of the murine glucose-6-phosphatase gene opens the way for studying the molecular basis of GSD type 1a in humans and its etiology in an animal model. PMID- 8407996 TI - Inhibition of thapsigargin-induced calcium entry by microinjected guanine nucleotide analogues. Evidence for the involvement of a small G-protein in capacitative calcium entry. AB - Injection of mouse lacrimal acinar cells with the non-hydrolyzable analogue of GTP, GTP gamma S (guanosine 5'-3-O-(thio)triphosphate), caused a rapid release of intracellular calcium but failed to activate calcium entry. Injection of GTP gamma S together with the inositol 1,4,5-trisphosphate receptor inhibitor, heparin, did not induce calcium release but blocked the activation of capacitative calcium entry by thapsigargin. Injection of GDP beta S (guanosine 5' O-(thio)diphosphate) produced similar effects. The inhibitor effects of GDP beta S were prevented if an equal concentration of GTP was included in the injection pipette. These findings suggest that one of the steps linking the depletion of intracellular calcium pools to calcium entry across the plasma membrane requires the hydrolysis of GTP and may involve a small G-protein. PMID- 8407997 TI - Factor VII autoactivation proceeds via interaction of distinct protease-cofactor and zymogen-cofactor complexes. Implications of a two-dimensional enzyme kinetic mechanism. AB - Tissue factor (TF), an integral membrane protein, enhances the feedback activation of factor VII by factor VIIa (factor VII autoactivation). We found that, in contrast to the other known membrane-dependent coagulation activation reactions, TF-dependent factor VII autoactivation occurred preferentially on neutral phospholipid vesicles relative to negatively charged vesicles containing phosphatidylserine. This reaction was best described by a novel mechanism in which the enzyme and substrate are each bound to separate cofactor (TF) molecules. This unusual mechanism of substrate presentation to a membrane-bound protease predicts that the reaction rate will be directly dependent on the surface density, and hence lateral diffusion, of factor VII.TF and factor VIIa.TF complexes, obeying obligatorily two-dimensional enzyme kinetics. This prediction was confirmed, yielding a two-dimensional second-order rate constant (k2D) of 4.9 (+/- 0.8) x 10(6) m2 mol-1 s-1. Since intact cells normally sequester acidic phospholipids away from the outer leaflet of the plasma membrane, this reaction mechanism should permit factor VII autoactivation to predominate on unactivated/undamaged cell surfaces when other clotting reactions are dormant. PMID- 8407998 TI - Conversion of a magnesium binding site into a zinc binding site by a single amino acid substitution in Escherichia coli alkaline phosphatase. AB - The replacement of aspartic acid by histidine at position 153 in Escherichia coli alkaline phosphatase results in a mutant enzyme that is remarkably similar to certain mammalian alkaline phosphatases that are activated by magnesium in a time dependent fashion. These mammalian alkaline phosphatases have histidine at the position corresponding to 153 of the E. coli sequence. Here we report the three dimensional structure of the mutant E. coli alkaline phosphatase with histidine at position 153. The structure reveals that the octahedral magnesium binding site has been converted to a tetrahedral zinc binding site with an imidazole ring nitrogen of His-153 as one of the ligands to the zinc. The alteration in metal binding caused by the mutation could explain the origin of the magnesium activation observed with the mammalian alkaline phosphatases. The structure also reveals differences in the mode of phosphate binding, explaining the enhanced phosphate affinity and the reduced activity of the mutant enzyme in the presence of zinc. PMID- 8407999 TI - Site-directed mutagenesis of the phosphorylation sites in the mouse glucocorticoid receptor. AB - The functional significance of receptor phosphorylation in mediating the actions of glucocorticoids remains undefined. The identification of seven phosphorylation sites in the mouse glucocorticoid receptor (Bodwell, J. E., Orti, E., Coull, J. M., Pappin, D. J. C., Smith, L. I., and Swift, F. (1991) J. Biol. Chem. 266, 7549 7555) permits a direct examination of the potential regulatory role of glucocorticoid receptor phosphorylation in transactivation. Using oligonucleotide directed mutagenesis of the mouse glucocorticoid receptor cDNA, we have substituted alanine or aspartate for the residues phosphorylated in this ligand dependent transcription factor. COS-1 cells were cotransfected with mutant receptor cDNA expression vectors and a reporter plasmid containing the glucocorticoid-inducible mouse mammary tumor virus promoter linked to chloramphenicol acetyltransferase in order to characterize the effect of these substitutions on receptor-mediated gene expression. Substitution of alanine or aspartate at single phosphorylation sites does not prevent receptor transactivation. Receptors containing multiple substitutions of alanine or aspartate at the major phosphorylation sites in the acidic domain elicit levels of hormone-induced reporter gene expression that are comparable to wild-type receptors. Mutant receptors substituted with alanine at the five phosphorylation sites conserved among the rat, human, and mouse receptors exhibit a 22% decrease in transcriptional activity. Receptors mutated at all seven sites display a similar modest reduction. These results demonstrate that receptor phosphorylation at these seven identified residues is not a major determinant in glucocorticoid receptor transcriptional activity at the mouse mammary tumor virus promoter. PMID- 8408000 TI - Neutrophil phospholipase D is activated by a membrane-associated Rho family small molecular weight GTP-binding protein. AB - Phospholipase D in human neutrophil lysates is activated by GTP gamma S (guanosine 5'-O-(3-thiotriphosphate)), implying the participation of a GTP binding protein. Reconstitution of GTP gamma S-dependent activity requires protein factors in both the plasma membrane and the cytosol (Olson, S. C., Bowman, E. P., and Lambeth, J. D. (1991) J. Biol. Chem. 266, 17236-17242). The location of the GTP-binding protein was investigated by preincubating either cytosol or plasma membrane with GTP gamma S, followed by removal of all but tightly bound nucleotide and reconstituting activity with the complementing untreated subcellular fraction. This approach indicated that the GTP-binding protein was membrane-associated. A low magnesium requirement for GTP gamma S prebinding, as well as a failure of aluminum fluoride to activate, suggested a Ras-like small M(r) GTP-binding protein. smg GDP dissociation stimulator, which stimulates the exchange of GDP for GTP on a variety of small GTP-binding proteins, stimulated GTP-dependent phospholipase D activity. Rho GDP dissociation inhibitor, a regulatory protein that binds specifically to and inhibits the functions of Rho family small GTP-binding proteins, inhibited GTP gamma S dependent activity. Thus, neutrophil phospholipase D is regulated by a membrane associated small molecular weight GTP-binding protein, likely to be a member of the Rho family. PMID- 8408001 TI - Characterization of hypoxia-inducible factor 1 and regulation of DNA binding activity by hypoxia. AB - Hypoxia-inducible factor 1 (HIF-1) is a DNA binding activity detected in nuclear extracts from Hep3B cells cultured in 1% O2 but not in extracts from cells cultured in 20% O2. HIF-1 binds to a sequence within the human erythropoietin gene enhancer that is required for hypoxic activation of transcription. Induction of HIF-1 is inhibited by cycloheximide, which also inhibits induction of erythropoietin RNA. We now demonstrate that induction of both HIF-1 and erythropoietin RNA is inhibited by the protein kinase inhibitor 2-aminopurine. HIF-1 binding to DNA was eliminated by phosphatase treatment of nuclear extracts. Actinomycin D also inhibited HIF-1 induction, suggesting that de novo transcription is required. The kinetics of HIF-1 induction by hypoxia paralleled the kinetics of erythropoietin gene transcriptional induction. HIF-1 DNA binding activity decayed rapidly when hypoxic cells were exposed to increased oxygen tension. In vitro, the kinetics of HIF-1 association with, and dissociation from, its binding site were extremely rapid, with a t1/2 for both processes of < 1 min. These findings are consistent with the proposed function of HIF-1 as a physiologic regulator of gene expression that responds to changes in cellular oxygen tension. Methylation interference analysis indicated that HIF-1 makes specific contacts with DNA in the major groove. PMID- 8408002 TI - Phospholipase D hydrolysis of choline phosphoglycerides is selective for the alkyl-linked subclass of Madin-Darby canine kidney cells. AB - Madin-Darby canine kidney (MDCK) cells were used to study the synthesis of diglycerides from choline phospholipids (PC) in response to 12-O tetradecanoylphorbol-13-acetate (TPA). In this system, diglyceride formation was blocked in the presence of ethanol (0.5%), and a corresponding amount of phosphatidylethanol (PEt) was formed, indicating that phospholipase D is responsible for the diglyceride production. Analysis of the subclasses of phosphatidylethanol revealed 1-O-alkyl-(alkyl), 1-O-alk-1'-enyl-(alkenyl), and 1 acyl species of PEt (38.0, 8.3, and 53.7%, respectively). The molecular species of the alkyl-PEt most closely matched the alkyl-PC. No change in the relative amounts of alkyl- versus acyl-PEt was observed with time after stimulation. Comparison of the alkyl content of PEt (38.0%) and the parent PC (15.2%) indicated a marked selectivity for the alkyl subclass of PC. A cell-free assay (Huang, C., Wykle, R. L., Daniel, L. W., and Cabot, M. C. (1992) J. Biol. Chem. 267, 16859-16865) for phospholipase D was also used to confirm the selectivity of the enzyme for alkyl-PC versus acyl-PC. The predominant molecular species of PEt contained saturated acyl or alkyl chains in position-1 and monounsaturated residues in position-2 accounting for approximately 50% of the total PEt. 1-O Octadecyl-2-oleoyl-sn-glycerol, a representative alkyl molecular species, was synthesized and tested for its effect upon protein kinase C derived from MDCK cells. This alkyl-diglyceride (DG) neither stimulated protein kinase C nor inhibited its activation by diacylglycerol. In summary, TPA-stimulated phospholipase D is selective for the alkyl-PC subclass in MDCK cells. The alkyl DG subsequently formed does not appear to function as a second-messenger in activating protein kinase C. PMID- 8408003 TI - Interleukin-11, an inducible cytokine in human articular chondrocytes and synoviocytes, stimulates the production of the tissue inhibitor of metalloproteinases. AB - This study on the regulation of interleukin (IL)-11 expression in human connective tissue cells shows that IL-11 expression is not restricted to cells of hematopoietic origin but can also be induced in articular chondrocytes and synoviocytes. IL-11 mRNA was induced in chondrocytes in response to transforming growth factor (TGF)-beta 1 and IL-1 beta. Stimulation with IL-6 or growth factors, such as basic fibroblast growth factor, leukemia inhibitory factor, and platelet-derived growth factor-AA, had only weak or no detectable effects. Activation of protein kinase C by phorbol esters and inhibition of protein synthesis by cyclohexamide increased IL-11 transcripts, whereas calcium ionophore A23817 or dibutyryl cyclic AMP had no effect. Immunoprecipitations revealed the synthesis of IL-11 protein in response to TGF-beta 1, IL-1 beta, as well as phorbol 12-myristate 13-acetate, and a synergistic action of TGF-beta 1 and IL-1 beta was observed. Similar findings on IL-11 expression were made in synoviocytes. Analysis of effects on cell function showed that IL-11 stimulated the production of the tissue inhibitor of metalloproteinases in chondrocytes and synoviocytes but did not affect chondrocyte proliferation or increase stromelysin activity. These results suggest that IL-11 does not contribute to connective tissue degradation but conversely induces protective effects in joint tissue. PMID- 8408004 TI - Intrinsic stoichiometric equilibrium constants for the binding of zinc(II) and copper(II) to the high affinity site of serum albumin. AB - Intrinsic stoichiometric equilibrium constants were determined for zinc(II) and copper(II) binding to bovine and human serum albumin. Data were obtained from equilibrium dialysis experiments. Metals were presented to apoprotein as metal chelates in order to avoid metal hydrolysis and to minimize nonspecific metal protein interactions. Scatchard analysis of the binding data indicated that the high affinity class for both zinc and copper was comprised of one site. Results of binding experiments done at several pH values suggested that while both histidyl and carboxyl groups appear to be involved in copper binding, histidyl residues alone were sufficient for zinc binding. These amino acid residues were used in combination to model several binding sites used in the formulation of equilibria expressions from which stoichiometric constants were calculated. The log10K for bovine serum albumin were calculated to be 7.28 for Zn(II) and 11.12 for Cu(II). Those for human serum albumin were determined to be 7.53 and 11.18 for Zn(II) and Cu(II), respectively. These constants were used in equilibria to simulate speciation of metal-albumin and metal-chelator and to illustrate relative binding affinities. This comparison of binding strengths was possible only through the calculation of an intrinsic stoichiometric binding constant. PMID- 8408005 TI - Expression of cellular retinoic acid-binding protein (type II) in Escherichia coli. Characterization and comparison to cellular retinoic acid-binding protein (type I). AB - Cellular retinoic acid-binding protein type II, CRABP(II), has been expressed efficiently in Escherichia coli from the mouse cDNA and compared to E. coli expressed cellular retinoic acid-binding protein type I, CRABP(I). CRABP(II) had a molecular weight approximately 15,700, a pI of 5.46, an Amax of 350 nm with A350/A280 of 1.8, a fluorescence excitation maximum at 347 nm, and a fluorescence emission maximum at 465 nm (holoprotein). All-trans-retinoic acid and 3,4 didehydro-, 4-hydroxy-, 4-oxo-, 16-hydroxy-4-oxo-, and 18-hydroxy-retinoic acids bind CRABP(II) and CRABP(I) stoichiometrically (Kd values no greater than approximately 10-20 nM). 9-cis-Retinoic acid exhibited saturation binding to both CRABP(II) and CRABP(I) with similar affinities, Kd = approximately 50-70, as did 13-cis-retinoic acid, Kd = approximately 160-240 nM, demonstrating that CRABP(II) and CRABP(I) have similar orders of selectivity for known retinoids. HoloCRABP(II) transferred approximately 70% of its all-trans-retinoic acid to CRABP(I), however, indicating that CRABP(II) has approximately a 3-fold lower affinity for stoichiometrically binding retinoids than CRABP(I). Stern-Volmer plots of both native and denatured CRABP(II) and CRABP(I) were consistent with comparable loci for the tryptophan residues and similar three-dimensional structures. These results suggest that CRABP(II) and CRABP(I), when expressed in vivo in excess of total retinoids, bind several ligands, and functional dissimilarities between the two proteins would not be related to unique preferences for known endogenous retinoids. Rather, CRABP(I) and CRABP(II) may modulate the steady-state concentrations of retinoids to different set points. PMID- 8408006 TI - Different biochemical modes of action of two irreversible H+,K(+)-ATPase inhibitors, omeprazole and E3810. AB - Omeprazole and E3810 were found to inhibit gastric H+,K(+)-ATPase following different biochemical mechanisms. Effects of the specific binding of the inhibitors on the conformational state of the enzyme were studied by measuring the fluorescence of the enzyme labeled with fluorescein 5'-isothiocyanate. The absolute fluorescence level of the omeprazole-bound enzyme was lower than that of the control enzyme, and reduction of S-S cross-linking between the enzyme and omeprazole increased the fluorescence. Addition of K+ into the control vesicle solution quenched the fluorescence (E1-->E2K+). The quench was inhibited in the omeprazole-bound enzyme but not in the E3810-bound enzyme. These results suggest that the omeprazole-bound enzyme has a low fluorescence conformation (E2 form). On the other hand, the conformation of the E3810-bound enzyme was the same as that of the control enzyme (E1 form). Phosphoenzyme formation in the absence of K+ was inhibited in both the E3810- and omeprazole-bound enzymes. Binding of 2',3'-o-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate to the enzyme was equally inhibited by E3810 and omeprazole. K(+)-dependent dephosphorylation from the phosphoenzyme was inhibited in the E3810-bound enzyme but not in the omeprazole bound enzyme. These experimental results have shown that the inhibition mechanism of H+,K(+)-ATPase by omeprazole was different from that by E3810; the partial reaction that was the most differently affected by the inhibitors was the conformational change from the E2 to E1 form for omeprazole and the luminal K(+) dependent dephosphorylation for E3810. PMID- 8408007 TI - Isolation and characterization of an intracellular serine proteinase inhibitor from a monkey kidney epithelial cell line. AB - A recent report described a thrombin inhibitory activity in the soluble fraction of human placenta and the cytosolic fraction of K562 cells. Isolation and characterization of the functionally inactive 35-38-kDa placental form of this protein revealed that it was a novel serine proteinase inhibitor (Coughlin, P. B., Tetaz, T., and Salem, H. H. (1993) J. Biol. Chem. 268, 9541-9547). In the present study, we observed a 67-kDa sodium dodecyl sulfate (SDS)-stable complex when 125I-thrombin was incubated with the cytosolic fraction of a monkey kidney epithelial cell line, BSC-1. This complex was not observed in either the particulate cell fraction extracted with 0.2% Triton X-100 or medium conditioned by cells, suggesting that the thrombin-complexing factor is confined to the cytoplasm. The cytoplasmic antithrombin activity was purified to apparent homogeneity from the cytosol of BSC-1 cells previously pulsed with [35S]methionine by a combination of heparin-agarose chromatography, Mono Q fast protein liquid chromatography, and anhydrotrypsin-Affi-Gel 10 affinity chromatography. Analysis of the affinity-purified preparation by SDS polyacrylamide gel electrophoresis and fluorography revealed a single protein with an apparent molecular mass of 38 kDa. The purified 38-kDa protein inhibited the amidolytic activities of thrombin, trypsin, urokinase, and factor Xa but not that of elastase. Incubation of the 38-kDa protein with excess thrombin identified approximately 60% of the labeled 38-kDa protein in an SDS-stable 67 kDa complex. The purified 38-kDa inhibitor was cleaved with cyanogen bromide and the isolated peptides subjected to microsequencing. Amino acid sequence obtained for a region within this protein exhibited significant homology with human antithrombin III and plasminogen activator inhibitors 1 and 2. This homologous peptide contained the full complement of residues designated as highly conserved in helix F of the greater serine proteinase inhibitor superfamily. In addition, an internal sequence of GGGGDIHQGF was found in the monkey cytoplasmic inhibitor, which is identical to that reported for an internal sequence of the human placental inhibitor. These findings confirm the existence of a novel cytoplasmic serine proteinase inhibitor in mammalian cells and provide additional details of its molecular properties. The physiological function of this novel serine proteinase inhibitor in cytoplasm is unknown. PMID- 8408008 TI - Transient intermediates of the methane monooxygenase catalytic cycle. AB - Three new intermediates of the catalytic cycle of the soluble form of methane monooxygenase (MMO) isolated from Methylosinus trichosporium OB3b have been detected using transient kinetic techniques. MMO consists of hydroxylase (MMOH), reductase, and "B" (MMOB) components. MMOH contains an oxygen-bridged [Fe(III).Fe(III)] cluster that catalyzes O2 activation and insertion chemistry. At 4 degrees C, rapid mixing of O2 with diferrous MMOH ([Fe(II).Fe(II)]) in the presence of a 2-fold excess of MMOB resulted in loss of the g = 16 EPR signal characteristic of the diferrous cluster at an apparent first order rate of 22 +/- 5 s-1 (O2 approximately 700 microM). Subsequently, an EPR silent, chromophoric (lambda max = 330 and 430 nm, epsilon approximately 7500 M-1 cm-1 at each wavelength) intermediate (compound Q) formed with an average first order rate constant of 1 +/- 0.1 s-1 and then decayed at 0.05 +/- 0.01 s-1. Since compound Q formed much more slowly than diferrous MMOH disappeared, at least one other undetected intermediate (compound P) must have formed before compound Q. MMO substrates had little or no effect on the formation rate of compound Q, but they caused the decay rate to increase linearly with the concentration added. The substrates methane, furan, and nitrobenzene caused compound Q decay to occur with second order rate constants of 19,000 M-1 s-1, 9000 M-1 s-1, and 200 M-1 s-1 (+/- 5%), respectively. When nitrobenzene was used as a substrate, a second chromophoric intermediate (compound T, lambda max = 325 nm, with a shoulder at 395 nm, epsilon 395 approximately 6000 M-1 cm-1) formed at the same rate as compound Q decay. Chemical quench studies showed that compound T is an enzyme product complex that decays with a rate constant of 0.02 +/- 0.005 s-1. This rate is approximately the same as kcat for nitrobenzene turnover at 4 degrees C catalyzed by the reconstituted MMO system, suggesting that product release is the rate-limiting step in catalysis. The characteristics of compound Q suggest that it may be the activated form of the enzyme that directly catalyzes substrate oxidation. PMID- 8408009 TI - Enzymes converting D-3-hydroxyacyl-CoA to trans-2-enoyl-CoA. Microsomal and peroxisomal isoenzymes in rat liver. AB - Epiermization of 3-hydroxyacyl-CoA, which has been shown to occur as a two-step dehydration-hydration reaction (Hiltunen, J. K., Palosaari, P. M., and Kunau, W. H. (1989) J. Biol. Chem. 264, 13536-13540; Smeland, E., Jianxun, L., Chu, C., Cuebas, D., and Schulz, H. (1989) Biochem. Biophys. Res. Commun. 160, 988-992) was studied in rat liver. Subcellular fractionations of rat liver on different density gradients revealed a dual distribution of activity, catalyzing dehydration of D-3-hydroxydecanoyl-CoA to trans-2-decenoyl-CoA (hydratase 2) in both peroxisomal and microsomal compartments. Both hydratase 2 activity peaks were separated by dye ligand chromatography from the extract of the combined heavy and light mitochondrial fractions. The activity eluted at low salt was identified as the microsomal isoform and was purified to apparent homogeneity. The M(r) of the native protein (subunit) was found to be 60,000 (31,500), indicating that it is homodimeric. The enzyme activity was inhibited by IgGs isolated from antisera raised against the denatured subunit. The activity eluted at high salt was tentatively identified to be peroxisomal of origin, and the M(r) of the native protein (subunit) was determined to be 62,000 (33,500). The peroxisomal enzyme was not recognized by the antibody to its microsomal counterpart. Analysis of the reaction products of microsomal enzyme activity by gas chromatography-mass spectrometry showed that the enzyme catalyzed reversibly hydration/dehydration between trans-2-enoyl-CoA and D-3-hydroxyacyl-CoA, but L-3 hydroxydecanoyl-CoA was not dehydrated to delta 2-enoyl-CoA compounds. Similar reaction characteristics were also determined for the peroxisomal hydratase by using stereospecific auxiliary enzymes. The present data demonstrate that rat liver contains microsomal and peroxisomal proteins possessing hydratase 2 activities. Although their kinetic properties are similar, immunological data, subunit sizes, and chromatographic evidence clearly indicate that they are different enzymes. Comparisons with other hydratases revealed that the microsomal and peroxisomal hydratase 2 described in the present work are proteins that have not been previously purified. PMID- 8408010 TI - Redox, transferrin-independent, and receptor-mediated endocytosis iron uptake systems in cultured human fibroblasts. AB - Sepharose beads bound to 55Fe-transferrin (Tf) were used to evaluate Tf-dependent iron uptake not employing receptor-mediated endocytosis (RME). The iron of 55Fe2 Tf-Sepharose was reduced and taken up by cultured human fibroblasts in a time- and concentration-dependent fashion (Km 7 microM; Vmax 128 pmol/mg/min). This redox system resembled that for Tf-independent iron uptake (Tf-IU, evaluated using 55Fe-citrate) in several ways. 1) NH4Cl did not inhibit iron uptake from 55Fe-citrate and 55Fe2-Tf-Sepharose but did inhibit uptake from 55Fe2-Tf (RME system). 2) Iron uptake and reduction from 55Fe2-Tf-Sepharose and 55Fe-citrate increased with temperature hyperbolically, differing from the sigmoidal curve for RME uptake. 3) The subcellular distributions of iron from 55Fe-citrate and 55Fe2 Tf-Sepharose resembled each other and differed from that for 55Fe2-Tf. 4) The optimal pH for iron reduction and uptake using 55Fe2-Tf-Sepharose or 55Fe-citrate was less than pH 5.5, while that for iron uptake from 55Fe2-Tf was pH 7.4. 5) The uptake and reduction of iron from 55Fe2-Tf-Sepharose was inhibited by ferric citrate and by transition metals. We conclude that both Tf-independent and non RME, Tf-dependent iron uptake proceed via a common redox system for iron. The mechanisms of cellular iron uptake can be separately evaluated in fibroblasts using 55Fe-citrate, 55Fe2-Tf, and 55Fe2-Tf-Sepharose beads. PMID- 8408011 TI - Cloning and sequence analysis of the cDNA for arginine kinase of lobster muscle. AB - Arginine kinase belongs to an evolutionary conserved family of ATP:guanidino phosphotransferases, whose members play an important role in energy metabolism. In this work, a lambda gt11 lobster muscle library was constructed and screened by using both polyclonal antibodies and two synthetic oligonucleotides. The complete amino acid sequence of arginine kinase (ATP:L-arginine N phosphotransferase, EC 2.7.3.3) from lobster muscle was determined by cloning and sequencing the DNA complementary to its mRNA. The identity of the clone was confirmed by comparing the amino acid sequence deduced by nucleotide sequence analysis with previously published partial sequences of amino- and carboxyl terminal regions of the enzyme and some of its fragments (Regnouf, F., Kassab, R., Debuire, B., Richard, C., and Han K. K. (1981) Int. J. Peptide Protein Res. 17, 143-155). The nucleotide sequence of the cDNA was found to contain an open reading frame encoding 355 amino acid residues with a calculated molecular mass of 39,830 daltons. This enzyme exhibits a significant sequence identity to those of representative members of the guanidino kinase family, including creatine kinase. This report represents the first molecular cloning and sequencing of an ATP-guanidino phosphotransferase which is not a creatine kinase isoform. PMID- 8408012 TI - The transcription factor GATA-1 regulates the promoter activity of the platelet glycoprotein IIb gene. AB - Glycoprotein IIb (GPIIb) is an early and specific marker of the megakaryocytic lineage. We have previously shown that a fragment extending 643 base pairs upstream the transcription start site of the human GPIIb promoter was able to control the tissue-specific expression of the CAT gene in transfection experiments. Four potential GATA-binding sites, located at positions -463, -376, 243, and -54 are present within this fragment. Gel shift analysis revealed that nuclear extracts from the erythroleukemic cell line HEL contain a DNA-binding protein that recognizes these GATA sites. Using an antiserum raised to an hydrophilic region of the transcription factor GATA-1, the HEL GATA-binding protein was found to be GATA-1. Point mutations of the different GATA sites indicated that they did not equally contribute to GPIIb promoter activity. The 463 GATA motif located in an enhancer region is essential for full transcription activity and was found to be dominant upon the other GATA motifs. When this site is mutated, the -54 GATA site appears to be essential for the remaining CAT activity. These results indicate that the transcription factor GATA-1 plays an important role in the regulation of the transcription of the megakaryocyte specific GPIIb gene. PMID- 8408013 TI - Binding of lactoferrin and transferrin to the human promonocytic cell line U937. Effect on iron uptake and release. AB - We have compared the ability of lactoferrin and transferrin to interact with and donate iron to the monocytic cell line U937. About 10 times more lactoferrin was bound than transferrin, but most lactoferrin bound nonspecifically, and the degree of specific binding was similar for both proteins (2-3 x 10(6) sites/cell). The binding affinity for lactoferrin (83 nM) was about 4-fold lower than for transferrin (21 nM). Lactoferrin did not inhibit binding of transferrin, or vice versa. Binding of lactoferrin was not inhibited by 30 mM glucose or fucose nor by incubating the cells with heparinase. Transferrin, but not lactoferrin, was internalized, and 3 mM primaquine caused intracellular accumulation of transferrin but not lactoferrin. The cells rapidly acquired iron from transferrin, but uptake from lactoferrin was 10-fold slower and probably resulted from transfer of 59Fe from lactoferrin to unlabeled transferrin during culture. Lactoferrin, but not transferrin, released iron to the extracellular medium when bound to U937 cells. Lactoferrin inhibited cellular uptake of iron from Fe-nitrilotriacetate but not from transferrin. It is concluded that transferrin, but not lactoferrin, acts as an iron donor to U937 cells. Lactoferrin may regulate uptake of potentially toxic non-transferrin-bound iron. PMID- 8408014 TI - Regulation of the human serotonin transporter. Cholera toxin-induced stimulation of serotonin uptake in human placental choriocarcinoma cells is accompanied by increased serotonin transporter mRNA levels and serotonin transporter-specific ligand binding. AB - Treatment of confluent cultures of JAR human placental choriocarcinoma cells with cholera toxin or forskolin for 16 h markedly stimulated (2.4-fold) serotonin transport activity in these cells. Cycloheximide, an inhibitor of protein synthesis or actinomycin D, an inhibitor of mRNA synthesis effectively blocked this stimulation. Northern blot analysis revealed that treatment with cholera toxin resulted in severalfold increase in the concentrations of the three mRNA species (6.8, 4.9 and 3.0 kilobases in size) which hybridized to the human placental serotonin transporter cDNA. Under similar conditions, the concentrations of the mRNA species which hybridized to the human placental taurine transporter cDNA or to the human beta-actin cDNA were not affected. Analysis of paroxetine-sensitive binding of the cocaine analog 2 beta carbomethoxy-3 beta-(4- [125I]iodophenyl)tropane to the membranes prepared from control and cholera toxin-treated cells indicated that the maximal binding capacity was increased 2.5-fold by cholera toxin, with no significant change in the binding affinity. Thus, stimulation of serotonin transporter activity in the placental choriocarcinoma cells following cholera toxin treatment is likely a result of an increase in cell surface density of the serotonin transporter protein as a consequence of increased steady state serotonin transporter mRNA levels. PMID- 8408015 TI - The dihydrofolate reductase domain of Plasmodium falciparum thymidylate synthase dihydrofolate reductase. Gene synthesis, expression, and anti-folate-resistant mutants. AB - A 693-base pair gene coding for the 27,132-dalton dihydrofolate reductase (DHFR) domain of the thymidylate synthase-dihydrofolate reductase (TS-DHFR) bifunctional protein of Plasmodium falciparum was designed to have Escherichia coli codon preference and multiple unique restriction sites and was chemically synthesized. The gene was overexpressed (> 50% total cellular protein) in E. coli as insoluble inclusion bodies which could be unfolded and refolded to recover soluble enzyme activity. The refolded DHFR was purified by methotrexate-Sepharose affinity chromatography to give the homogeneous enzyme. Active site titration with methotrexate revealed that the purified protein was fully active. The purified DHFR migrates as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with apparent mass of approximately 30 kDa, and gel filtration showed that the protein is a monomer. The yield of purified enzyme was about 5-6 mg/liter of bacterial culture. Kinetic properties of the purified recombinant DHFR were similar to those reported for wild type bifunctional TS-DHFR. Cassette mutagenesis of the synthetic gene was performed to give the S108N and the N51I + S108N mutants which provided DHFRs analogous to pyrimethamine-resistant mutants found in nature. PMID- 8408016 TI - Utility of polyhistidine-tagged ubiquitin in the purification of ubiquitin protein conjugates and as an affinity ligand for the purification of ubiquitin specific hydrolases. AB - The purification and biochemical characterization of protein substrates of the ubiquitin-dependent pathway of proteolysis is made difficult in part by the low steady state levels of ubiquitin-protein conjugates. We report here on the use of a polyhistidine-tagged ubiquitin molecule (HisUb) for the purification of ubiquitin-protein conjugates by metal chelate chromatography. When Escherichia coli extracts containing expressed HisUb were passed through a nitrilotriacetic acid-agarose column containing immobilized Ni2+ ions (Ni-NTA column), HisUb was retained. After washing to remove unbound and nonspecifically bound proteins, a pH 4.5 wash was used to elute highly purified HisUb. Purified HisUb and wild-type ubiquitin were tested for their ability to form Ni(2+)-binding ubiquitin-protein conjugates in a wheat germ in vitro conjugation reaction. In some experiments, wheat germ extracts were preincubated with iodoacetamide to inhibit ubiquitin activating and conjugating enzymes. Only those conjugation assays containing HisUb and an ATP-regenerating system not pretreated with iodoacetamide produced significant levels of multiple Ni(2+)-binding ubiquitin-protein conjugates. We also examined the potential of HisUb as an affinity ligand for the purification of higher plant ubiquitin-specific hydrolases. As a test, a crude lysate of E. coli expressing a yeast ubiquitin-specific hydrolase (Yuh1) was passed through a Ni-NTA column containing bound HisUb. Yuh1 was retained on the column and was specifically eluted when the column was equilibrated with buffer containing wild type ubiquitin. PMID- 8408017 TI - Effects of beta 6 aromatic amino acids on polymerization and solubility of recombinant hemoglobins made in yeast. AB - Valine, leucine, tryptophan, and phenylalanine substitutions at the beta 6 position of hemoglobin (Hb) were made using a yeast expression system coupled with a polymerase chain reaction-based mutagenesis strategy. The oxygen affinity and absorption spectra of these mutants were similar to recombinant Hb A except for Hb beta E6W which had a higher absorbance at approximately 280 nm. The deoxy forms of Hb beta E6L and Hb S showed characteristic delay times prior to polymerization. Tetrameric deoxy-Hbs containing tryptophan or phenylalanine at the beta 6 position had higher solubilities and polymerized less readily compared with deoxy-Hb S. However, when oversaturated, these Hbs polymerized without a delay time. These results suggest that Hb beta E6W and Hb beta E6F form polymers upon deoxygenation by a linear polymerization mechanism without nuclei formation. During polymerization, bulky hydrophobic amino acids, like phenylalanine and tryptophan at the beta 6 position, might interact with the acceptor pocket on the surface of an adjacent Hb molecule but may not be able to form stable hydrophobic interactions like beta 6 valine and leucine. Difficulty in insertion of the bulky side chains of these aromatic amino acids into the hydrophobic acceptor pocket on an adjacent tetramer may inhibit nuclei formation prior to polymerization. PMID- 8408018 TI - Regulation of folate and one-carbon metabolism in mammalian cells. I. Folate metabolism in Chinese hamster ovary cells expressing Escherichia coli or human folylpoly-gamma-glutamate synthetase activity. AB - Chinese hamster ovary (CHO) cell transfectants expressing various levels of human and Escherichia coli folylpoly-gamma-glutamate synthetase (FPGS) activity and possessing different folylpolyglutamate chain length distributions have been developed as models for folate and antifolate metabolism. The synthesis of pteroyltriglutamate was sufficient for normal cellular retention of folate and also overcame the phenotypic requirement for purines and thymidine of AUXB1, a CHO cell mutant lacking FPGS activity and lacking folylpolyglutamates. Only low levels of FPGS are required to enable cellular metabolism of folates to forms that are retained by mammalian cells. The higher levels found in mammalian cells are required for the synthesis of the long chain polyglutamate derivatives characteristic of mammalian cells. At low medium folate concentrations, folate accumulation by transfectants expressing human FPGS was not responsive to FPGS levels as the limiting step in metabolism was beyond the triglutamate, the chain length required for retention. The rate-limiting step in folate metabolism in cells expressing the E. coli enzyme was the conversion of diglutamate to triglutamate, and, at low FPGS levels, the E. coli enzyme was about 50-fold less effective than the human FPGS in enabling cellular folate accumulation. These data suggest that cellular accumulation of any folate analog whose mono- or diglutamate derivative is a poor substrate for FPGS would be very responsive to the level of FPGS activity. PMID- 8408019 TI - Regulation of folate and one-carbon metabolism in mammalian cells. II. Effect of folylpoly-gamma-glutamate synthetase substrate specificity and level on folate metabolism and folylpoly-gamma-glutamate specificity of metabolic cycles of one carbon metabolism. AB - The effect of folylpoly-gamma-glutamate synthetase (FPGS) levels on folate accumulation was investigated in Chinese hamster ovary cells expressing various levels of human and Escherichia coli FPGS activity. At low medium folate concentrations, folate accumulation was limited by influx and was independent of FPGS activity except in cells expressing extremely low levels of FPGS. Essentially all transported folate was metabolized to retained polyglutamate derivatives, the chain length of which varied with the level of FPGS activity. As medium folate concentration increased through the physiological to the pharmacological range, cellular folate accumulation became proportional to FPGS activity and the chain length of intracellular folates decreased. At high folate concentrations, competition between substrates for FPGS limited the extent of polyglutamylation and less than 5% of transported folate was retained by the cell. Pteroyltriglutamates functioned as effectively as the longer chain length polyglutamates normally found in mammalian cells in the metabolic cycles of de novo purine and thymidylate biosynthesis but were unable to support glycine and methionine synthesis. Transfectants expressing human FPGS and containing folates of glutamate chain length ranging from four to eight were equally effective at supporting glycine synthesis, and transfectants expressing higher levels of FPGS were able to grow in the absence of methionine. Growth in the absence of methionine required high (nonphysiological) intracellular folate levels and longer chain length polyglutamates. PMID- 8408020 TI - Regulation of folate and one-carbon metabolism in mammalian cells. III. Role of mitochondrial folylpoly-gamma-glutamate synthetase. AB - Wild-type Chinese hamster ovary (CHO) cells and CHO cell transfectants expressing human folylpoly-gamma-glutamate synthetase (FPGS) activity contain mitochondrial FPGS activity of higher specific activity than the cytosolic isozyme. Expression of mitochondrial FPGS activity is required for folate accumulation by mitochondria. The mitochondrial folate pool in CHO cells is not in equilibrium with the cytosolic pool and contains folylpolyglutamates of longer glutamate chain length than cytosolic folates. The inability of AUX-coli, a CHO cell expressing high levels of Escherichia coli FPGS activity and containing pteroyltriglutamate, to support glycine synthesis is due to a lack of mitochondrial FPGS activity. AUX-coli cells lack mitochondrial folate despite containing high levels of cytosolic folate. PMID- 8408021 TI - Regulation of folate and one-carbon metabolism in mammalian cells. IV. Role of folylpoly-gamma-glutamate synthetase in methotrexate metabolism and cytotoxicity. AB - Chinese hamster ovary (CHO) cells expressing human and Escherichia coli folylpoly gamma-glutamate synthetase (FPGS) activities were used as models to study factors regulating the cytotoxicity and metabolism of methotrexate (MTX). CHO cells expressing human FPGS metabolized MTX to polyglutamates characteristic of human cells. Cellular MTX accumulation and metabolism to polyglutamates were dependent on the level of FPGS activity and were unaffected by putative gamma-glutamyl hydrolase inhibitors. The sensitivity of cells continuously exposed to MTX was not influenced by FPGS activity. After short term exposure to MTX, cells expressing higher levels of FPGS were more sensitive to the drug. MTX was not transported into the mitochondria and MTX treatment had no effect on preexisting mitochondrial folates while cytosolic folates were converted to oxidized forms. Mitochondrial folate accumulation was significantly impaired by MTX treatment, suggesting that the mitochondrial folate transport system is specific for reduced folates. PMID- 8408022 TI - YTRF is the conserved internalization signal of the transferrin receptor, and a second YTRF signal at position 31-34 enhances endocytosis. AB - By functional analysis of mutant human transferrin receptors (TR) expressed in chicken embryo fibroblasts, we previously identified a tetrapeptide sequence, Y20TRF23, within the 61-residue cytoplasmic tail as the signal for high efficiency endocytosis (Collawn, J. F., Stangel, M., Kuhn, L. A., Esekogwu, V., Jing, S., Trowbridge, I.S., and Tainer, J.A. (1990) Cell 63, 1061-1072). It has been inferred from other studies, however, that the TR internalization signal was localized to a much larger region, residues 7 through 26 (Girones, N., Alvarez, E., Seth, A., Lin, I-M., Latour, D.A., and Davis, R.J. (1991) J. Biol. Chem. 266, 19006-19012). Additionally, Tyr20 was reported to not be conserved in the Chinese hamster cytoplasmic tail sequence (Alvarez, E., Girones, N., and Davis, R.J. (1990) Biochem. J. 267, 31-35). In the studies reported here, we examined the effect of insertion of an extra copy of a YTRF sequence at three different locations within the human TR cytoplasmic domain and show that the insertion of another YTRF signal at position 31-34 in the wild-type TR, but not the other two locations, increases the rate of endocytosis 2-fold. Furthermore, introduction of YTRF at position 31-34 in an internalization-defective mutant receptor restores endocytosis to wild-type levels, indicating that YTRF signals at either positions 20-23 or 31-34 are necessary and sufficient to promote TR internalization and function in an independent and additive manner. We also report the complete primary structure of the Chinese hamster TR deduced from its cDNA sequence and show that the Tyr20 as well as the complete YTRF motif is conserved. PMID- 8408023 TI - (S)-geranylgeranylglyceryl phosphate synthase. Purification and characterization of the first pathway-specific enzyme in archaebacterial membrane lipid biosynthesis. AB - The first pathway-specific step in the biosynthesis of the core membrane diether lipids in archaebacteria is the alkylation of the primary hydroxyl group in (S) glyceryl phosphate by geranylgeranyl diphosphate. The reaction is catalyzed by (S)-3-O-geranylgeranylglyceryl phosphate ((S)-GGGP) synthase. The cytosolic enzyme was purified to homogeneity from the moderately thermophilic archaebacterium Methanobacterium thermoautotrophicum by a combination of ammonium sulfate precipitation, four chromatographic steps (DE52, Q-Sepharose, phenyl Superose, and Protein Pak), and native polyacrylamide gel electrophoresis. SDS polyacrylamide gel electrophoresis of gel-purified GGGP synthase gave a single band at 29 kDa. The enzyme requires Mg2+ for optimal activity, although prenyltransfer is also seen in buffers containing Mn2+ or Zn2+. A well defined pH optimum occurs between 6.0 and 7.5. Maximal activity is seen at 50-65 degrees C. The Michaelis constants for GGGP synthase are Vmax = 4.1 +/- 0.5 mumol min-1 mg 1, KMGGPP = 4.1 +/- 1.1 microM, and KMGP = 41 +/- 5 microM. PMID- 8408024 TI - Raf exists in a native heterocomplex with hsp90 and p50 that can be reconstituted in a cell-free system. AB - Recently, we have demonstrated that the tyrosine kinase pp60v-src can undergo cell-free assembly into a heterocomplex with rabbit hsp90 and p50 when the immunoadsorbed protein is incubated with rabbit reticulocyte lysate (Hutchison, K. A., Brott, B. K., De Leon, J. H., Perdew, G. H., Jove, R., and Pratt, W. B. (1992) J. Biol. Chem 267, 2902-2908). Using a baculovirus system to express a high level of human c-Raf serine/threonine kinase in Sf9 insect cells, we show here that immunoadsorbed c-Raf undergoes similar lysate-mediated assembly into a heterocomplex with hsp90 and p50. As with pp60v-src and steroid receptors, binding of c-Raf to hsp90 occurs in an ATP-dependent and K(+)-dependent manner and the resulting heterocomplex is stabilized by molybdate. With a very rapid and gentle procedure of Sf9 cell cytosol preparation and c-Raf immunoadsorption, we show coimmunoadsorption of the insect homologue of hsp90. The same procedures permit detection of a native complex of v-Raf with rat hsp90 and p50 in stably transfected rat 3Y1 fibroblasts, and v-Raf is also assembled into a heterocomplex with rabbit hsp90 and p50 by reticulocyte lysate. Using the 22W mutant of c-Raf in which the NH2-terminal half has been deleted, we show that the catalytic domain of the kinase is sufficient for both formation of the native heterocomplex in mouse NIH 3T3 cells and cell-free reconstitution of the heterocomplex by rabbit reticulocyte lysate. Although the native Raf-heat shock protein heterocomplex is less stable than native pp60v-src and glucocorticoid receptor heterocomplexes, by analogy with these proteins its detection may have important implications regarding the mechanism of Raf trafficking through the cytoplasm. PMID- 8408025 TI - Initiation of glycogen synthesis. Control of glycogenin by glycogen phosphorylase. AB - Glycogen biosynthesis involves a specific initiation event, mediated by a specialized protein, glycogenin. Glycogenin undergoes self-glucosylation to generate an oligosaccharide primer, which, when long enough, supports the action of glycogen synthase to elongate the polysaccharide chain, leading ultimately to the formation of glycogen. We report that primed glycogenin is also a substrate for glycogen phosphorylase. Phosphorylase removed glucose from the oligosaccharide attached to glycogenin in a phosphorolysis reaction that required phosphate and produced glucose 1-phosphate. The phosphorylated form, phosphorylase a, was much more effective than the dephosphorylated phosphorylase b. However, in the presence of the allosteric effector AMP, phosphorylase b also catalyzed the phosphorolysis reaction. Glucose, an allosteric inhibitor of phosphorylase, inhibited the reaction. Glycogen, but not a short oligosaccharide (maltopentaose), also inhibited the reaction. Treatment of fully primed glycogenin with phosphorylase converted the glycogenin to a form with slightly lower apparent molecular weight, which was less effective as a substrate for glycogen synthase. These results suggest a novel role for phosphorylase in the control of glycogen biosynthesis. We propose that the glucosylation level of glycogenin would be determined by the balance between the self-glucosylation reaction and the opposing action of phosphorylase. The level of glucosylation would in turn determine whether or not glycogenin was an effective primer for glycogen synthase. In this way, several known controls of phosphorylase activity, such as epinephrine, glucagon, and insulin, could influence not only the elongation/degradation stage of glycogen metabolism but also its initiation. PMID- 8408026 TI - Phenobarbital induction and tissue-specific expression of the rat CYP2B2 gene in transgenic mice. AB - To investigate molecular events regulating the transcription of genes inducible by phenobarbital, transgenic mouse strains were developed incorporating rat cytochrome P450 2B2 (CYP2B2) genes. Expression in mouse tissues was analyzed for two series of rat CYP2B2 gene constructs, of 19 and 39 kilobase pairs total length, each containing the entire coding region, introns, and 3'-flanking sequences of CYP2B2, but differing in the respective lengths of 5'-flanking sequence. One group of mice, whose transgene included the complete 2B2 gene but only 800 base pairs of 5'-proximal sequence, were not phenobarbital-inducible in mouse liver or in any extrahepatic tissue; rather, these genes were expressed at very high levels constitutively and selectively in only kidney and liver. A second group of mice with an identical transgene, except for the presence of an additional 19 kilobase pairs of 5'-flanking sequence, expressed 2B2 only in the liver and at high levels only after phenobarbital treatment, analogous to the expression pattern observed for the endogenous CYP2B2 gene in the rat. These results demonstrate that, in vivo, phenobarbital induction and tissue-specific control requires interaction of regulatory elements far upstream of the core CYP2B2 promoter region and upstream of motifs indicated previously as determinants of phenobarbital responsiveness. PMID- 8408027 TI - N-acetylglucosaminyltransferase regulates the expression of neolactoglycolipids including sulfoglucuronylglycolipids in the developing nervous system. AB - Lactosylceramide N-acetylglucosaminyltransferase (GlcNac-Tr) in the synthesis of lactotriosylceramide (LcOse3Cer) was characterized in the nervous system. The microsomal membrane GlcNAc-Tr required a divalent metal ion, preferably Mn2+, and a nonionic detergent. The pH optimum was around 7.0. The enzyme also transferred GlcNAc to neolactotetraosylceramide (nLcOse4Cer), GM1, and asialo-GM1, but not to other glycolipids. The Km value for lactosylceramide was 21 microM (Vmax = 91 pmol/mg/h), and that for nLcOse4Cer was 35 microM (Vmax = 112 pmol/mg/h). The GlcNAc-Tr for the glycolipids appears to be separate from that for oligosaccharides. The developmental expression of GlcNAc-Tr, both in the cerebral cortex and cerebellum, correlated well with the tissue levels of LcOse3Cer, nLcOse4Cer, sulfoglucuronylglycolipids (SGGLs), and other neolacto series glycolipids (nLSGs). In the cerebral cortex, the specific activity of GlcNAc-Tr decreased sharply from a maximum level at embryonic day 15, and by postnatal day 10 onward, it was undetectable. In the adult cerebral cortex, although significant activities of other glycosyltransferases involved in the subsequent steps of the synthesis of SGGLs were present, the absence of GlcNAc-Tr stymied the formation of LcOse3Cer and therefore the synthesis of nLSGs, including SGGLs. In the cerebellum, the GlcNAc-Tr specific activity declined from the day of birth to postnatal day 3, but later, the activity increased and reached a maximum at postnatal day 15, which correlated with the increasing synthesis of nLSGs. The results indicate that lactosylceramide GlcNAc-Tr is the key regulatory enzyme controlling the differential expression of all nLSGs in the developing nervous system. PMID- 8408028 TI - A deletion mutation in the third cytoplasmic loop of the mouse m1 muscarinic acetylcholine receptor unmasks cryptic G-protein binding sites. AB - Mutations were introduced in the highly conserved carboxyl-terminal region of the third cytoplasmic loop of the mouse m1 muscarinic acetylcholine receptor (mAChR) gene by site-directed mutagenesis. The effects of these mutations on ligand binding and on mAChR coupling to phosphoinositide turnover have been examined following expression in mouse Y1 adrenal carcinoma cells, Chinese hamster ovary (CHO) cells, and Rat-2 fibroblasts. Point mutations in the region proximal to the sixth transmembrane domain had no effect on antagonist binding but did result in decreased agonist affinity. A deletion of four amino acids in the same region effectively uncoupled the receptor from phosphoinositol turnover in Y1 adrenal carcinoma cells but had no effect in either CHO cells or Rat-2 fibroblasts. Differential sensitivity to pertussis toxin indicates that the m1 mAChR can interact with multiple G-proteins in CHO cells and Rat-2 cells via distinct recognition sites on the receptor. These data demonstrate multiple G-proteins can interact with an individual receptor, that the same receptor may couple to different second messenger pathways, and that these responses can vary in a cell type-specific manner. PMID- 8408029 TI - Endopeptidase from rat liver membranes, which generates miniglucagon from glucagon. AB - An endopeptidase activity that cleaves glucagon, producing miniglucagon or glucagon (19-29), a Ca2+ pump inhibitory peptide, was isolated from rat liver membranes. The purified enzyme has a molecular mass of approximately 100 kDa and a pH optimum of approximately 8. It is inhibited by both sulfhydryl-blocking reagents and metal-chelating reagents and activated by thiol compounds. The partial N-terminal amino acid sequence of the 100-kDa protein does not correspond to any known protein. An antiserum was raised against a synthetic octapeptide corresponding to the N-terminal sequence. Immunoblot analysis of crude liver membranes revealed a single band at 100 kDa. Immunoreactivity was found in liver, pancreas, and heart, which are glucagon and miniglucagon target tissues, and in gastric mucosa and kidney. Low levels were detected in spleen, whereas immunoreactivity was undetectable in skeletal muscle and intestinal mucosa. The endopeptidase activity was inhibited by insulin, glucagon-like peptide-1, and glucagon-like peptide-1 (7-36) amide, whereas other peptides that contain dibasic sites had no effect on its activity, indicating that the endopeptidase does not display strict selectivity toward basic doublets. PMID- 8408030 TI - Histone H1-mediated inhibition of transcription initiation of methylated templates in vitro. AB - The detailed mechanism underlying the inhibition of transcription by DNA methylation is still obscure. Chromatin structure has frequently been proposed as a role player in this mechanism. Histone H1 is a known key element in the formation and stabilization of chromatin fibers. We describe here experiments designed to examine the effect of DNA methylation on the binding of histone H1 to DNA and the consequent inhibitory effect of the bound histone H1 on in vitro transcription. The results of these experiments showed a clear preferential binding of histone H1 to methylated DNA as compared with unmethylated DNA. The in vitro transcription assay indicated that transcription of methylated templates was inhibited at a lower histone H1/DNA ratio than of unmethylated templates, and that the extent of inhibition depends on the density of methyl groups in the promoter region. This inhibition of in vitro transcription was alleviated efficiently by methylated competitor DNA, whereas, under similar conditions, almost no effect was observed with unmethylated competitor. Experiments designed to pinpoint the stage in the transcription process that was suppressed by the preferred binding of histone H1 to methylated template revealed that inhibition occurred at the initiation and not at the elongation level. PMID- 8408031 TI - A revised structure for fucoidan may explain some of its biological activities. AB - Fucoidan from Fucus vesiculosus inhibits human sperm-zona pellucida binding and blocks the zona pellucida-induced acrosome reaction in human sperm. Fucoidan also potently inhibits selectin-mediated adhesion of leukocytes to vascular endothelium. To understand the molecular basis for fucoidan's inhibition of specific cell adhesion events, we have investigated the structure of this fucan using definitive methods of carbohydrate structural analysis. We report the revised average structure for fucoidan. [formula: see text] This average structure differs from the previous model for fucoidan in two respects. First, the core region of the fucan is composed primarily of a polymer of alpha 1-3 linked fucose with sulfate groups substituted at the 4 position on some of the fucose residues. Secondly, fucose is also attached to this polymer to form branch points, one for every 2-3 fucose residues within the chain. This revised average structure is consistent with previous studies suggesting a branched random coil as the best model for this polysaccharide. The proposed model is also a closer structural analogue of the sulfated carbohydrate ligands that bind to selectins. This information should be useful for determining whether a relationship exists between selectin-mediated adhesion of leukocytes and human sperm-egg binding. PMID- 8408032 TI - Ethanol inhibits the autophosphorylation of the insulin-like growth factor 1 (IGF 1) receptor and IGF-1-mediated proliferation of 3T3 cells. AB - The effect of ethanol on cell proliferation was studied in Balb/c 3T3 cells and in stably transfected 3T3 cells constitutively overexpressing the human insulin like growth factor 1 (IGF-1) receptor (p6 cells). Ethanol inhibited growth of both cell lines when they were cultured in serum-free medium supplemented with individual growth factors, i.e. platelet-derived growth factor and IGF-1 for 3T3 cells, and IGF-1 only for p6 cells. Increases in cell number were prevented in both cell lines even when ethanol was present exclusively during the period of IGF-1 stimulation. The inhibitory effect of ethanol was concentration-dependent, with a 30% inhibition observed at 10 mM ethanol. IGF-1 receptor tyrosine autophosphorylation was completely prevented by ethanol both in intact cells and in immunopurified IGF-1 receptor preparations. The binding of IGF-1 to its receptor on intact cells was unaffected by ethanol. Ethanol also inhibited the stimulation of IGF-1 receptor autophosphorylation and the corresponding growth of p6 cells induced by IGF-2. Transcription of c-myc, c-fos, and c-jun in response to IGF-1 was inhibited by ethanol. These findings demonstrate that ethanol at low concentrations markedly inhibits IGF-1 receptor autophosphorylation and IGF-1 mediated cell growth. PMID- 8408033 TI - Purification and characterization of a DNA helicase from Saccharomyces cerevisiae. AB - A novel DNA helicase, scHelI, has been purified from whole cell extracts of Saccharomyces cerevisiae using biochemical assays to monitor the fractionation. The enzyme unwinds partial duplex DNA substrates, as long as 343 base pairs in length, in a reaction that is dependent on either ATP or dATP hydrolysis. scHelI also catalyzes a single-stranded DNA-dependent ATP hydrolysis reaction; the apparent Km for ATP is 325 microM. The unwinding reaction on circular partial duplex substrates is biphasic, with a fast component occurring within 5 min of the initiation of the reaction and a slow component continuing to 60 min. This is in contrast to the ATP hydrolysis reaction, which exhibits linear kinetics for 60 min. The direction of the unwinding reaction is 5' to 3' with respect to the strand of DNA on which the enzyme is bound. The unwinding reaction is strongly stimulated by the addition of Escherichia coli single-stranded DNA-binding protein when long partial duplex substrates are used. The enzymatic activity of scHelI copurifies with a polypeptide of 135 kDa as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The polypeptide sediments as a monomer in a glycerol gradient in the presence of 0.2 M NaCl. PMID- 8408034 TI - Mechanism of action of Pseudomonas exotoxin. Identification of a rate-limiting step. AB - Pseudomonas exotoxin (PE) enters cells by receptor-mediated endocytosis and is cleaved by a cellular protease between Arg279 and Gly280 to produce an NH2 terminal fragment of 28 kDa which contains the toxin's binding domain and a COOH terminal fragment of 37 kDa which has translocating and ADP-ribosylating activity. After proteolysis, the COOH-terminal fragment reaches the endoplasmic reticulum by retrograde transport where it translocates to the cytosol and inhibits protein synthesis by ADP-ribosylating elongation factor 2. To understand how the 37-kDa fragment functions, we focused on the role of specific amino acids located near its NH2 terminus. We found that there was a 4-250-fold loss in toxic activity when tryptophan 281, leucine 284, or tyrosine 289 were changed to other residues. Mutations at these three positions did not interfere with the receptor binding, cell-mediated proteolytic cleavage, or ADP-ribosylating activity. To determine the role of these amino acids, a competition assay was devised in which the addition of excess PE delta 553, a mutant form of PE that lacks ADP ribosylation activity, competed efficiently for the toxicity of PE. Excess PE with mutations near the NH2 terminus of the 37-kDa fragment competed poorly. This competition occurred after proteolysis since PEGly276, a mutant form of PE that is not cleaved, did not complete. We conclude that specific amino acids at the NH2 terminus of the 37-kDa fragment interact in a saturable manner with an unknown intracellular component. PMID- 8408035 TI - Skelemin, a cytoskeletal M-disc periphery protein, contains motifs of adhesion/recognition and intermediate filament proteins. AB - In striated muscle, myofibrils are anchored to an interconnecting cytoskeleton of desmin intermediate filaments. Skelemin (195 kDa) may be a link between myofibrils and the intermediate filament cytoskeleton. Skelemin partitions with desmin to the insoluble cytoskeleton, and increases the thickness of reconstituted intermediate filaments. Concentrated at the M-disc periphery, skelemin may also contact myosin filaments. We used immunoscreening to isolate a mouse muscle cDNA which encodes a protein with a calculated molecular mass of 185 kDa. Anti-skelemin antibodies bound to the protein products of each of three nonoverlapping regions of the open reading frame. Antibodies directed against the protein products of each one-third of the cDNA react with a 195-kDa muscle protein and stain the M-disc indistinguishably from the original anti-skelemin antibodies, suggesting that the cDNA encodes skelemin. A single skelemin mRNA is detected in muscle but not non-muscle tissues, consistent with immunostaining results. Skelemin is a member of a family of myosin-associated proteins containing fibronectin type III and immunoglobulin superfamily C2 motifs. Skelemin is unique in this family in having intermediate filament core-like motifs, one near each terminus. We hypothesize that skelemin could interact with myosin or myosin-associated proteins through its fibronectin and/or immunoglobulin motifs, and with intermediate filaments through intermediate filament-like motifs. PMID- 8408036 TI - Thrombospondin 1 is a tight-binding competitive inhibitor of neutrophil cathepsin G. Determination of the kinetic mechanism of inhibition and localization of cathepsin G binding to the thrombospondin 1 type 3 repeats. AB - Thrombospondin 1 was recently shown to bind to and inhibit the activity of neutrophil elastase (Hogg, P. J., Owensby, D. A., Mosher, D. F., Misenheimer, T. M., and Chesterman, C. N. (1993) J. Biol. Chem. 268, 7139-7146). This finding led us to question whether thrombospondin 1 also binds and inhibits the other major serine proteinase of neutrophils, cathepsin G. In a competitive binding assay, cathepsin G bound to thrombospondin 1 reversibly and saturably with a dissociation constant in the low nanomolar range. The kinetic mechanism of inhibition of cathepsin G activity by thrombospondin 1 was determined using the synthetic cathepsin G substrate, Suc-Ala-Ala-Pro-Phe-p-nitroanilide, and is consistent with hyperbolic tight-binding inhibition in which thrombospondin 1 binds cathepsin G and the Michaelis cathepsin G-substrate complex and weakens, but does not abolish, the efficiency of hydrolysis of Suc-Ala-Ala-Pro-Phe-p nitroanilide. In the presence of 2 mM calcium ions, 2.9 +/- 0.4 mol of cathepsin G interacted with 1 mol of thrombospondin 1 trimer with a site-binding constant of 7.0 +/- 3.5 nM, which reduced the efficiency of hydrolysis of Suc-Ala-Ala-Pro Phe-p-nitroanilide 8.5 +/- 1.4-fold. A lower limit for the on rate constant of 5 x 10(6) M-1 S-1 was established. The affinity of binding and stoichiometry for the interaction between cathepsin G and thrombospondin 1 was enhanced in the absence of calcium ions. In the presence of EDTA, 5.3 +/- 0.5 mol of cathepsin G interacted with 1 mol of thrombospondin 1 with a site-binding constant of 2.1 +/- 1.6 nM, implying the existence of two binding sites for cathepsin G on each subunit of thrombospondin 1, one or both of which is variably exposed and sensitive to calcium ions. Thrombospondin 1 protected fibronectin from cleavage by cathepsin G and blocked cathepsin G-mediated platelet aggregation. In summary, the binding of cathepsin G to thrombospondin 1 is tight, reversible, and close enough to the active site of cathepsin G to perturb the interactions of a small synthetic substrate and exclude a macromolecular protein substrate and platelets. Using defined proteolytic fragments and different conformers of thrombospondin 1, the binding sites for cathepsin G have been localized to the thrombospondin 1 type 3 repeats. PMID- 8408037 TI - Modulation of furin-mediated proprotein processing activity by site-directed mutagenesis. AB - The proprotein processing activity of mutants of the subtilisin-like enzyme furin was studied in transfected mammalian cells. Our studies indicate that the three residues of the catalytic triad of furin, Asp46, His87, and Ser261, are critical not only for substrate processing but also for maturation of furin. Furthermore, evidence is provided that maturation of furin occurs through an intramolecular autocatalytic process. Substitution of the asparagine residue (Asn188) of the oxyanion hole by an alanine residue appears to block substrate processing but not furin maturation. Analysis of carboxyl-terminal deletion mutants revealed that the segment encompassing residues Glu449 to Glu469 of the "middle" domain, which is more than 100 residues downstream of the predicted catalytic domain, contains residues that seem to be critical for processing activity but that the more carboxyl-terminal cysteine-rich region, the transmembrane region, and the cytosolic tail are dispensable. Finally, we made mutants in the substrate binding region of human furin and studied their ability to process von Willebrand factor (pro-vWF) substrates, including wild-type pro-vWF as well as pro-vWF mutants in which the P1 (vWFR-1G), P2 (vWFK-2A), or P4 (vWFR-4A) basic residue with respect to the pro region cleavage site had been mutated. It is demonstrated that particular negatively charged residues in or near the substrate binding region of furin are critical for cleavage activity and specificity of the enzyme for multiple basic residues in the substrate. Furthermore, substrate binding region mutants of furin were obtained, which cleaved either the pro-vWFK-2A or pro-vWFR 4A mutant of pro-vWF more efficiently than wild-type pro-vWF. PMID- 8408038 TI - Genomic organization and expression of two independent gene arrays coding for two antigenic acidic ribosomal proteins of Leishmania. AB - In the present paper we describe the isolation and characterization of four novel genes of the parasitic protozoan Leishmania infantum. These genes are organized as two independent gene clusters, and they are related by nucleotide sequence to eukaryotic genes encoding acidic ribosomal proteins. Each gene cluster contains two tandemly linked genes coding for identical proteins. Each of the proteins coded by the gene clusters (called LiP and LiP') are highly divergent in sequence, showing the characteristic features of eukaryotic P-proteins from the P2 group. In spite of the sequence conservation of the coding regions of each of the genes in the cluster, the 5'- and 3'-untranslated regions are heterogeneous in sequence. The analysis of the expression of these genes indicates that logarithmic phase promastigotes show increased levels of LiP- and LiP'-specific transcripts compared with stationary phase promastigotes. The steady state RNA levels of the LiP and LiP' genes show a similar dependence of the growth phase of the parasite. Using specific probes for the divergent 3'-untranslated regions of each of the genes, it was found that the abundance of the mature transcripts is different even when the transcripts are derived from the same gene cluster. These findings probably indicate that the 3'-untranslated regions may influence the stability or turnover of the transcripts derived from both LiP and LiP' gene clusters. PMID- 8408039 TI - Purification and characterization of VDE, a site-specific endonuclease from the yeast Saccharomyces cerevisiae. AB - The 119-kDa primary translation product of the VMA1 gene of Saccharomyces cerevisiae undergoes a self-catalyzed rearrangement ("protein splicing") that excises an internal 50-kDa segment of the polypeptide and joins the amino terminal and carboxyl-terminal segments to generate the 69-kDa subunit of the vacuolar membrane-associated H(+)-ATPase. We have shown previously that the internal segment is a site-specific endonuclease (Gimble, F. S., and Thorner, J. (1992) Nature 357, 301-306). Here we describe methods for the high level expression and purification to near homogeneity of both the authentic VMA1 derived endonuclease (or VDE) from yeast (yield 18%) and a recombinant form of VDE made in bacteria (yield 29%). Detailed characterization of these preparations demonstrated that the yeast-derived and bacterially produced enzymes were indistinguishable, as judged by: (a) behavior during purification; (b) apparent native molecular mass (50 kDa); (c) immunological reactivity; and (d) catalytic properties (specific activity; cleavage site recognition; and optima for pH, temperature, divalent cation and ionic strength). The minimal site required for VDE cleavage was delimited to a 30-base pair sequence within its specific substrate (the VMA1 delta vde allele). PMID- 8408040 TI - Autoregulation-deficient mutant of the plasmid R6K-encoded pi protein distinguishes between palindromic and nonpalindromic binding sites. AB - The autogenously regulated gene pir of Escherichia coli plasmid R6K encodes the replication protein pi. This protein binds to two sites in the operator region of the pir gene: a 22-base pair nonpalindromic sequence and a pair of palindromic 9 base pair sequences. These pi-binding sites are similar, suggesting that pi uses a single DNA-binding domain in recognizing them. We devised a plasmid system permitting isolation of mutants of the pi protein which are altered in autoregulation. A Ser87 to Asn substitution in one such mutant, designated pi 87, reduces the protein's ability to repress the pir gene promoter in vivo. DNase I protection and gel retardation assays were carried out with highly purified pi 87 protein. In these studies pi 87 exhibited altered binding to the palindromic but not to the nonpalindromic part of the operator of the pir gene. Chemical cross linking and gel filtration analyses have shown that the dimerization properties of wild type pi and pi 87 proteins are similar in solution. We propose that the interaction of pi protein with the palindromic part of the pir operator is essential for autoregulation; we also propose that there is a fundamental difference in the mechanisms of pi protein recognition of palindromic and nonpalindromic sequences. PMID- 8408041 TI - Splicing and 3'-processing of the tyrosine tRNA of Trypanosoma brucei. AB - Trypanosoma brucei belongs to a family of protozoa that is characterized as having diverged early in the evolution of eukaryotes. Many unusual forms of RNA processing have been discovered in these cells, one of which is trans-splicing, and, although a great number of genes have been sequenced, no evidence for cis splicing has yet been found. This study shows that tRNA(Tyr) of T. brucei contains an 11-nucleotide intron. tRNA of the size predicted for unspliced precursor was detected by Northern analysis using an oligonucleotide probe complementary to the putative intervening region. Direct sequence analysis of mature tRNA(Tyr) showed that the predicted intron region is absent from this form of the molecule. Mutated versions of the gene encoding tRNA(Tyr) were introduced into trypanosomes by DNA transformation and shown to be expressed in vivo. Introduction into the genome of a tRNA(Tyr) gene containing an amber suppressor mutation led to a significant accumulation of unspliced precursor molecules. Analysis of the tRNA(Tyr) species from both the wild type and transformed cells revealed unspliced and spliced processing intermediates. The relative abundance of each of these intermediates suggests that 3'-processing and splicing occur independently and that, in the wild type, splicing tends to precede 3'-processing and CCA addition. PMID- 8408042 TI - Antisense inhibition of group II phospholipase A2 expression blocks the production of prostaglandin E2 by P388D1 cells. AB - Macrophage-like P388D1 cells release [3H]arachidonic acid and produce prostaglandin E2 (PGE2) upon stimulation with bacterial lipopolysaccharide (LPS) and platelet-activating factor (PAF). To determine whether group II phospholipase A2 (PLA2) is involved in this release, we treated P388D1 cells with antisense inhibitors specific for group II PLA2 RNA. Treatment with oligonucleotide ASGII decreased PLA2 activity in P388D1 cell homogenates by approximately 60% and reduced the release of [3H]arachidonic acid and PGE2 from activated cells to nearly resting cell levels. The inhibition by antisense oligonucleotide ASGII was blocked when its sense complement, SGII, was included in the incubation mixture. Stably transfected P388D1 cells expressing an antisense construct for group II PLA2 also produced reduced quantities of PGE2 in response to LPS and PAF. These data suggest that prostaglandin production by activated P388D1 cells involves phospholipid hydrolysis by group II PLA2. Oligonucleotide ASGII also blocked the appearance of a heparin-releasable group II PLA2 in the culture supernatants of P388D1 cells. The disappearance of this protein correlated with reduced PGE2 production by activated cells, indicating that an extracellular heparin associated pool of group II PLA2 is involved in prostaglandin production by P388D1 cells. PMID- 8408043 TI - Random mutagenesis of G protein alpha subunit G(o)alpha. Mutations altering nucleotide binding. AB - Nucleotide binding properties of the G protein alpha subunit G(o)alpha were probed by mutational analysis in recombinant Escherichia coli. Thousands of random mutations generated by polymerase chain reaction were screened by in situ [35S]GTP gamma S (guanosine 5'-(3-O-thio)-triphosphate) binding on the colony lifts following transformation of bacteria with modified G(o)alpha cDNA. Clones that did not bind the nucleotide under these conditions were characterized by DNA sequence analysis, and the nucleotide binding properties were further studied in crude bacterial extracts. A number of novel mutations reducing the affinity of G(o)alpha for GTP gamma S or Mg2+ were identified. Some of the mutations substitute amino acid residues homologous to those known to interact with guanine nucleotides in p21ras proteins. Other mutations show that previously unstudied residues also participate in the nucleotide binding. Several mutants lost GTP gamma S binding but retained the capacity to interact with the beta gamma subunit complex as determined by pertussis toxin-mediated ADP-ribosylation. One of these, mutant S47C, was functionally expressed in Xenopus laevis oocytes along with the G protein-coupled thyrotropin-releasing hormone (TRH) receptor. Whereas wild-type G(o)alpha increased TRH-promoted chloride currents, S47C significantly decreased the hormone-induced Cl- response, suggesting that this mutation resulted in a dominant negative phenotype. PMID- 8408044 TI - Overexpression in Escherichia coli and purification of an ATP-binding peptide from the yeast plasma membrane H(+)-ATPase. AB - The two major hydrophilic domains from the Saccharomyces cerevisiae plasma membrane H(+)-ATPase fused to glutathione S-transferase have been expressed in Escherichia coli. The GST-L peptide contained the hydrophilic region from Ala340 to Ser660. The GST-SL peptide contained in addition the hydrophilic region Glu162 to Val276. After solubilization of the inclusion bodies with urea, renaturation, and affinity chromatography, 3 mg of highly purified peptides were recovered per liter of E. coli culture. The purified peptides interacted with 2'(3')-O-(2,4,6 trinitrophenyl)-adenosine-5'-triphosphate (TNP-ATP), the fluorescence of which was enhanced identically upon binding of either GST-L or GST-SL. ATP competitively displaced the TNP-ATP binding. The observed dissociation constants for TNP-ATP (6.5 microM) and ATP (3 mM) are close to those found for the complete native H(+)-ATPase protein. The fluorescence of TNP-ATP was sensitive to Mg2+ indicating the existence of a Mg(2+)-binding site on the peptide. Apparent affinity for this Mg2+ site was found to vary from 50 microM at pH 7.5 to 400 microM at pH 5.5. PMID- 8408045 TI - Phosphorylation of dystrophin. The carboxyl-terminal region of dystrophin is a substrate for in vitro phosphorylation by p34cdc2 protein kinase. AB - In this paper, we report that p34cdc2 protein kinase phosphorylates recombinant fragments of skeletal muscle dystrophin with a maximal incorporation of 1.8 mol of Pi/mol of protein. Phosphorylation of both serine and threonine residues occurs within the carboxyl-terminal 201 amino acids of dystrophin, with phosphothreonine localized to within 25 residues of the carboxyl terminus. Supporting these in vitro studies, we also show that native dystrophin is phosphorylated by p34cdc2 kinase in isolated sarcolemmal vesicles. Sequence analysis indicates two consensus sites for p34cdc2 protein kinase within the carboxyl-terminal 201 amino acids of dystrophin. Importantly, neither of these sites is conserved in dystrophin-related protein, and only one site is conserved in the 71-kDa alternative product of the Duchenne muscular dystrophy gene, despite an otherwise extremely high degree of sequence conservation between these proteins. Importantly, in this study we also show that dystrophin is phosphorylated in vivo in rat skeletal muscle primary cultures, and we suggest that further investigation of both in vivo and in vitro phosphorylation of this protein will comprise an important part in determination of its function(s). PMID- 8408046 TI - Matrix metalloproteinase-3 (stromelysin-1). Identification as the cartilage acid metalloprotease and effect of pH on catalytic properties and calcium affinity. AB - Human pro-MMP-3 (pro-matrix metalloproteinase-3) was purified from three sources: articular cartilage and conditioned media from synovial fibroblasts and Chinese hamster ovary cells expressing recombinant pro-MMP-3. All three preparations reacted with two monoclonal antibodies specific for human fibroblast pro-MMP-3. Each preparation of active MMP-3 possessed properties identical to those previously reported for the cartilage acid metalloproteinase (MMP-6; Azzo and Woessner, J. F., Jr. (1986) J. Biol. Chem. 261, 5434-5441): an acid pH optimum of 5.3-5.5 for digestion of cartilage aggrecan; digestion of oxidized insulin B chain at Ala14-Leu15 and Tyr16-Leu17 in a ratio of 3:1; and heat stability at neutral pH. Further characterization of MMP-3 establishes that the acid pH optimum for cartilage aggrecan is not due to substrate denaturation since the same optimum is found by viscosity assay, by SDS-polyacrylamide gel electrophoresis assay of G1 domain, and by digestion of aggrecan in fresh cartilage fragments in vitro. Fibronectin was also digested optimally at pH 5.5 and NH2-terminal sequence analysis revealed no pH change in a major proteolytic site of cleavage at the Pro689-Leu690 bond. The specificity constant kcat/Km is maximal at pH 5.5 as determined in a quenched fluorescence peptide assay. This is due to an increase in kcat at pH 5.5 without any substantial effect on Km. The affinity of MMP-3 for calcium is decreased about 10-fold at pH 5.3 compared to neutral pH. Finally, the neutral cartilage metalloproteinase is identified as 72 kDa pro-MMP-2 based on M(r), specificity of insulin B-chain cleavage, and reactivity with a specific polyclonal antibody to human MMP-2. PMID- 8408047 TI - Negative autoregulation of c-Myb activity by homodimer formation through the leucine zipper. AB - The trans-activating and transforming capacities of the c-myb proto-oncogene product (c-Myb) are negatively regulated through a leucine zipper structure in its negative regulatory domain. We show here tht in cotransfection assays, maximal Myb-induced trans-activation occurs with relatively low amounts of wild type c-Myb, while higher levels of c-Myb result in reduced Myb-induced trans activation. By contrast, this apparent negative autoregulation is not observed with a c-Myb mutant containing an impaired leucine zipper. Data presented here suggest that this negative autoregulation of trans-activation by wild-type c-Myb is a consequence of homodimer formation by c-Myb through its leucine zipper and of the inability of c-Myb dimers to bind DNA. These findings point to a novel mechanism of regulation of a transcription factor. PMID- 8408048 TI - Temperature-sensitive phenotype of a mutant Sendai virus strain is caused by its insufficient accumulation of the M protein. AB - We investigated the process interrupting the production of a temperature sensitive mutant strain of Sendai virus, Cl.151, at the nonpermissive temperature (38 degrees C). The amount of virus M protein increased up to 6-fold when the cells persistently infected with Cl.151 strain at 38 degrees C are transferred to 32 degrees C, while the amount of nucleocapsid proteins did not alter. Cl.151 strain could restore virus production at 38 degrees C not only by the supplementation of M protein of wild type (Z) strain but also by the supplementation of M protein of Cl.151 strain. Neither the amount of M mRNA nor the rate of synthesis of M protein was altered by temperature in cells infected with the Cl.151 strain. However, we found that M protein of Cl.151 virus, which has 3-amino acid alterations from the wild type, was highly unstable at 38 degrees C when expressed under the control of an actin promoter. These results clearly show that Sendai virus M protein has a critical role in the production of virus particles without affecting virus gene expression. PMID- 8408049 TI - Transcriptional regulation of the rat prostaglandin endoperoxide synthase 2 gene in granulosa cells. Evidence for the role of a cis-acting C/EBP beta promoter element. AB - The promoter of the rat prostaglandin endoperoxide synthase 2 (PGS-2) gene has recently been shown to confer gonadotropic hormone (follicle-stimulating hormone (FSH), luteinizing hormone (LH), cAMP) inducibility when ligated to a CAT (chloramphenicol acetyltransferase) reporter gene and transfected into primary cultures of differentiated granulosa cells. To delineate cis-acting elements and trans-activating factors mediating this response, deletions of the active promoter region (-192/-53 base pairs upstream of the transcriptional start site) were tested for their ability to bind protein of granulosa cell nuclear extracts and activate reporter gene activity. Electrophoretic mobility shift assays revealed that the DNA subregion -142/-120 inhibited protein/DNA binding observed between granulosa cell nuclear extracts and the labeled PGS-2 fragment -192/-53. The subregion -142/-120 acting element C/EBP beta, 5'-TTATGCAAT-3'. Point mutations within the C/EBP beta element abolished protein/DNA binding and resulted in a 50% loss of forskolin/LH/FSH inducibility of reporter gene activity. C/EBP beta mRNA and protein were induced rapidly in granulosa cells in vivo by an ovulatory dose of human chorionic gonadotropin (hCG). Collectively, these results indicate that C/EPB beta appears to play a key role in regulating induction of the PGS-2 gene in granulosa cells prior to ovulation. PMID- 8408050 TI - Physiological effectors modify voltage sensing by the cyclosporin A-sensitive permeability transition pore of mitochondria. AB - This paper reports an investigation on the modulation of the mitochondrial permeability transition pore (MTP) by the membrane potential. Energized rat liver mitochondria loaded with a small Ca2+ pulse in sucrose medium supplemented with phosphate favor a high MTP "closed" probability because of the high membrane potential and therefore maintain a low permeability to sucrose. Upon depolarization by the addition of fully uncoupling concentrations of carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) mitochondria favor a high MTP "open" probability and rapidly undergo a process of osmotic swelling following sucrose diffusion toward the matrix. A titration with FCCP reveals that discrete subpopulations of mitochondria with different gating potentials for MTP opening may exist, since increasing concentrations of FCCP increase the fraction of mitochondria undergoing osmotic swelling. We show that physiological effectors (Ca2+, Mg2+, ADP, palmitate) modify pore opening in a mitochondrial population by shifting the fraction of mitochondria with a functionally open pore at any given membrane potential. Many inducers and inhibitors may therefore affect the pore directly through an effect on the MTP voltage sensing rather than indirectly through an effect on the membrane potential. Thus, many effectors may induce pore opening by shifting the MTP gating potential to higher levels, whereas many inhibitors may induce pore closure by shifting the MTP gating potential to lower levels. PMID- 8408051 TI - Purification of SEF1 proteins binding to transcriptional enhancer elements active in T lymphocytes. AB - Binding sites for SL3-3 enhancer factor 1 (SEF1) are important for the transcriptional activity in T lymphocytes and the tumorigenicity of SL3-3 murine leukemia virus. SEF1 is also implicated in the activity of many other leukemia, lymphoma, and sarcoma virus enhancers, and enhancers of genes for T cell receptor CD3 subunits. We have purified several proteins binding to SEF1 sites from bovine thymus using a five-step purification procedure. The proteins migrated as 19 distinct bands representing molecular masses from 23 kDa to about 200 kDa in SDS polyacrylamide gel electrophoresis. Ten DNA binding proteins, with molecular masses between 23 and 67 kDa, could be isolated after separation by SDS polyacrylamide gel electrophoresis. The DNA binding specificities of these proteins were similar and corresponded to that of the SEF1 binding activity in nuclear extracts. Each of these isolated SEF1 proteins also bound to the essential delta-E3 element of the human T cell receptor delta enhancer. Antibodies against one SEF1 protein only reacted with the protein used for immunization, which indicates a limited homology between at least some SEF1 proteins. We also present data suggesting that SEF1 proteins exist in multiple forms with differences in their DNA binding specificity, and that high affinity DNA binding of the SEF1 proteins requires protein phosphorylation. PMID- 8408052 TI - A cis-acting selector of a 5' splice site. Cooperation between the sequence of the site and an upstream exonic element. AB - To investigate the mechanism by which a 5' splice site (D1) is selected, while a nearby potentially functional site (Dcr1) is silenced, we have studied the importance of the 9-nucleotide sequence of these 5' splice sites for their respective usage. Our model system uses a transcript derived from the early transcription unit 3 of adenovirus-2. Transcripts, harboring an exonic element previously shown to be required for the selection of D1 in the presence of Dcr1, were mutated in the D1 and Dcr1 sequences and assayed for splicing in vitro. We first show that an increased ability of D1 to pair with U1 small nuclear (sn) RNA correlates with an increased accumulation of splicing intermediates, independently of the presence of Dcr1. This variation of efficiency of the first splicing reaction does not significantly affect the overall splicing efficiency except when the potential D1-U1 snRNA hybrid is less than 6 base pairs. Equally, the selector activity of the upstream exon element requires a D1 sequence that is able to form hybrids of 6 base pairs or more with U1 snRNA. This indicates that the cis-acting selector of D1 includes the exonic element (a potential stem-loop structure) and a D1 sequence of sufficient strength. PMID- 8408053 TI - Purification of chondroitin 6-sulfotransferase secreted from cultured chick embryo chondrocytes. AB - Chondroitin 6-sulfotransferase, which transfers sulfate from 3'-phosphoadenylyl sulfate to position 6 of N-acetylgalactosamine in chondroitin, was purified 1,430 fold to apparent homogeneity with a 22% yield from the serum-free culture medium of chick embryo chondrocytes by affinity chromatography on heparin-Sepharose CL 6B, wheat germ agglutinin-agarose, and 3',5'-ADP-agarose. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the purified enzyme showed a single broad protein band with an apparent molecular weight of 75,000. Since the purified enzyme has an apparent molecular weight of 160,000 as judged by gel chromatography on Superose 12, the active form of chondroitin 6-sulfotransferase may be a dimer. The purified enzyme transferred sulfate to chondroitin, chondroitin sulfate, and corneal keratan sulfate. Chondroitin sulfate E from squid cartilage, dermatan, sulfate, and heparan sulfate hardly served as acceptors of the sulfotransferase. The sulfated product derived from keratan sulfate was degraded by keratanase but not by chondroitinase ABC. PMID- 8408054 TI - Complete primary structure and biochemical properties of gilatoxin, a serine protease with kallikrein-like and angiotensin-degrading activities. AB - The activity and the complete primary structure of gilatoxin, a glycoprotein component from the venom of the Mexican beaded lizard (Heloderma horridum horridum) has been elucidated. Gilatoxin, a serine protease, showed kallikrein like activity, releasing bradykinin from kininogen; toxin-treated kininogen also produced lowered blood pressure in rats and contraction of isolated rat uterus smooth muscle. Gilatoxin catalyzed the hydrolysis of various arginine ester substrates for trypsin and thrombin and degraded both angiotensin I and II by cleavage of the dipeptide Asp-Arg from the NH2-terminal end. Fibrinogen was degraded but a fibrin clot was not produced, indicating that gilatoxin has specificities different from thrombin and snake venom thrombin-like proteases. The complete amino acid sequence of gilatoxin (245 residues) was deduced from NH2 terminal sequencing of overlapping peptide fragments cleaved from the reduced and alkylated toxin by enzymatic and chemical methods. The toxin is extensively glycosylated, containing approximately 8 mol of monosaccharide/mol of toxin, but appears to lack O-glycosylation sites. Amino acid sequence alignment of gilatoxin with batroxobin, crotalase, kallikrein, thrombin, trypsin, and several partial sequences of other Heloderma toxins reveals that there is considerable homology between these enzymes, particularly in the regions of the presumed catalytic site. Gilatoxin contains an additional 7 residues in the highly conserved catalytic region of serine proteases (including Asp-96, in the basic specificity pocket of thrombin) which may contribute to the unusual substrate specificity of the toxin. PMID- 8408055 TI - Effect of dexamethasone on the alternative splicing of the insulin receptor mRNA and insulin action in HepG2 hepatoma cells. AB - We have shown that culturing HepG2 cells in Ham's F-12 medium supplemented with calf serum, dexamethasone, and triiodothyronine causes an increase in the insulin sensitivity and responsiveness for glucose incorporation into glycogen. This correlates with increased expression of the mRNA encoding the B isoform of the insulin receptor. Of all the components in the medium, we found that dexamethasone exerted the greatest effect. Dexamethasone alone could cause both a switch in expression from the A to the B isoform of the insulin receptor and also an increase in insulin sensitivity for both glucose incorporation into glycogen and 2-deoxyglucose transport. In addition, we found that expression of the B isoform of the insulin receptor is developmentally regulated in the 3T3-L1 adipocyte leading to the suggestion that the alternatively spliced B isoform of the insulin receptor may play an important role in signal transduction in insulin target tissues such as liver, fat, and muscle. PMID- 8408056 TI - Cassette mutagenesis of a potential substrate recognition region of cytochrome P450 2C2. AB - Cassette mutagenesis was used to analyze the effects of mutations at amino acid positions 107-120 in cytochrome P450 2C2 which are part of a predicted substrate recognition site, SRS-1 (Gotoh, O. (1992) J. Biol. Chem. 267, 83-90), and terminate with a highly conserved tryptophan. All the mutant enzymes were expressed in COS1 cells at approximately the same level as wild type as determined by immunoprecipitation of radiolabeled products. Substitutions of amino acids from 107 to 110 and from 116 to 119 resulted in mutant enzymes that retained substantial lauric acid hydroxylase activity suggesting that these residues are not critical determinants of substrate specificity. In contrast, amino acids 112-115 were characterized by strong decreases in activity in at least one mutation tested. A substitution of phenylalanine for valine 112 reduced activity about 3-fold. At position 113, substitution of leucine for isoleucine had little effect, but activity was reduced 15-fold by substitution with cysteine. A conservative mutation at position 114 reduced activity 5-fold, a nonconservative mutation at position 115 resulted in a 30-fold reduction of lauric acid hydroxylation. Substitution of glutamic acid for glycine at position 109 had little effect on activity, while more conservative substitutions of valine for glycines at positions 111 and 117 resulted in about 50- and 7-fold reductions of activity, respectively. Substitution of leucine for tryptophan 120 resulted in a 7-fold reduction of activity indicating that tryptophan at this position contributes to, but is not essential for, enzymatic activity in spite of its presence in nearly all eukaryotic cytochromes P450. The results are consistent with a model in which amino acids 112-114, and possibly 115, align with a loop of bacterial P450cam sequence containing substrate contacting amino acids, tyrosine 96 and phenylalanine 98. Glycines at either end of the loop in cytochrome P450 2C2 appear to be critical for activity. PMID- 8408057 TI - Protein degradation by ERp72 from rat and mouse liver endoplasmic reticulum. AB - The endoplasmic reticulum (ER) resident protein, ER60, is a member of the protein disulfide-isomerase family and contains two copies of the internal thioredoxin motif, CGHC. Previously, ER60 was identified as a cysteine protease and named ER 60 protease (Urade, R., Nasu, M., Moriyama, T., Wada, K., and Kito, M. (1992) J. Biol. Chem. 267, 15152-15159; Urade, R., and Kito, M. (1992) FEBS Lett. 312, 83 86). Here, ERp72, the other member of the protein disulfide-isomerase family containing three CGHC motifs, was isolated from ER of rat and mouse livers through four sequential chromatographies on DEAE-Toyopearl 650, AF-heparin Toyopearl 650M, and TSK gel G3000SW twice. The purified rat protein was found to be homogeneous on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, not being contaminated by ER-60 protease, as judged on immunoblot analysis using an anti-ER-60 protease antibody. The partial amino acid sequence of rat ERp72 was 93% homologous to that of mouse ERp72. The purified rat ERp72 degraded other ER resident proteins such as protein disulfide-isomerase and calreticulin. The purified mouse ERp72 also degraded those proteins. Though rat ERp72 did not basically require Ca2+ for the reaction, the degradation of protein disulfide isomerase was enhanced, but the degradation of calreticulin was inhibited in the presence of Ca2+. The proteolytic activity of rat ERp72 was inhibited by cysteine protease inhibitors. Its sensitivity to protease inhibitors was the same as that of ER-60 protease. In addition, the proteolytic activity of rat ERp72 was inhibited by acidic phospholipids, also similar to ER-60 protease. Therefore, we propose that ERp72 be named ER-72 protease. PMID- 8408058 TI - Tyrosine phosphorylation of ras GTPase-activating protein does not require association with the epidermal growth factor receptor. AB - The importance of the carboxyl-terminal domain of the epidermal growth factor (EGF) receptor and its five autophosphorylation sites in the in vivo interaction and tyrosine phosphorylation of the ras GTPase-activating protein (rasGAP) has been investigated, using NIH 3T3 cells transfected with mutant EGF receptors. Phosphorylation of rasGAP by EGF receptor mutants, in which one to four autophosphorylation sites (Tyr-1173, -1148, -1086, and -1068) were mutated to phenylalanine, was reduced by 50-60% compared to the wild-type receptor. Elimination of these four autophosphorylation sites by truncation of 123 carboxyl terminal residues of the EGF receptor paralleled results obtained with point mutants. Substantial inhibition (about 90%) of rasGAP tyrosine phosphorylation by the EGF receptor occurred only when the remaining autophosphorylation site (Tyr 992) was mutated, in the context of this truncated receptor or in the full-length receptor mutated at all four other autophosphorylation sites. However, a point mutation of only Tyr-992 in the full-length receptor suppressed tyrosine phosphorylation of rasGAP only by 50%. In contrast, an EGF receptor lacking the last 214 amino acid residues (Dc214), which encompasses all five autophosphorylation sites, phosphorylated rasGAP to the same extent as the wild type receptor. However, this truncated receptor was significantly impaired in its capacity to phosphorylate phospholipase C-gamma 1. Interestingly, while EGF receptor autophosphorylation sites are required for EGF-induced rasGAP association with the receptor, maximal phosphorylation of rasGAP by the truncated receptor Dc214 occurred without detectable formation of receptor-rasGAP complexes. Furthermore, the capacity of mutated EGF receptors to bring about focal transformation was correlated with their capacity to phosphorylate rasGAP. PMID- 8408059 TI - RNA polymerase locations in the simian virus 40 transcription complex. AB - Transcription complexes of simian virus 40 can be isolated from cells late in infection in a form that retains the ability to continue transcription in vitro. These complexes have been investigated previously to gain information about the nucleoprotein structure of a transcribing gene. However, several studies have reported that the RNA polymerase molecules in such complexes are located almost entirely at the 5' end of the transcription unit. This would indicate that these complexes cannot be those which were in the process of transcribing mRNA in the cell, because the transcribing complexes would be expected to contain polymerase that were distributed throughout the transcription unit at the time of extraction. Since this issue is important to the interpretation of studies which characterize the extracted complexes, we have determined the polymerase locations using two new approaches. The first employs a new application of a procedure in which the transcription complex DNA is radioactively tagged by transcription in vitro. The second is a new method, which analyzes run-off transcripts generated in vitro. Both methods demonstrate that the polymerases are distributed throughout the genome in the transcription complexes. This result indicates that these complexes were in the process of transcribing mRNA at the time of cell lysis. PMID- 8408060 TI - 2,6-branched mannose and the regulation of poly-N-acetyllactosamine biosynthesis in N-linked oligosaccharides of Chinese hamster ovary cells. AB - Poly-N-acetyllactosamine (PL) sequences (repeating (3Gal beta 1-4GlcNAc beta 1)n) in complex-type N-linked oligosaccharides often occur in branched tri- and tetraantennary chains containing alpha-linked mannosyl residues disubstituted by N-acetyllactosaminyl units at C-2 and C-6 (2,6-branched mannose). We report here our studies on the factors affecting PL biosynthesis and the branching of N linked oligosaccharides in glycoproteins from Chinese hamster ovary (CHO) cells. For our studies, we utilized a mutant CHO cell line designated Lec8 CHO, which lacks the ability to galactosylate its glycoproteins and consequently synthesizes glycoproteins containing terminal GlcNAc residues and lacking poly-N-acetyl lactosamine and sialic acid. Glycoproteins in extracts of [3H]glucosamine- or [3H]mannose-labeled Lec8 CHO cells were galactosylated by exogenous beta 1-4 galactosyltransferase and analyzed by chromatography on leukoagglutinating phytohemagglutinin-Sepharose, a lectin reactive with glycoproteins containing 2,6 branched mannosyl residues. Approximately 10% of the radiolabeled glycoproteins were bound, and these were primarily of high molecular mass. Structural analyses of the bound glycoproteins demonstrated that they quantitatively contained 2,6 branched mannose. We then determined whether the "small i" N-acetylglucosaminyl transferase (iGNT), which initiates PL biosynthesis, could specifically recognize glycoproteins in vitro and whether recognition was dependent on the presence of 2,6-branched mannose. When the galactosylated glycoproteins in extracts of Lec8 CHO cells were incubated with UDP-[3H]GlcNAc, the endogenous iGNT quantitatively added GlcNAc in beta 1-3-linkage to terminal galactosyl residues in the leukoagglutinating phytohemagglutinin-bound glycoproteins. These results demonstrate for the first time that 2,6-branched mannosyl residues are restricted to a subset of CHO glycoproteins and that the iGNT in vitro preferentially recognizes glycoproteins containing the 2,6-branched mannose determinant. PMID- 8408061 TI - Biochemical evidence for a homophilic interaction of the alpha 3 beta 1 integrin. AB - The purpose of this study was to determine if the alpha 3 beta 1 integrin could interact in a homophilic manner. Several earlier reports have shown that certain integrin adhesion receptors, namely alpha 2 beta 1, alpha 3 beta 1, and alpha 6 beta 4 localize to intercellular adhesion structures and, therefore, may participate in cell-cell interactions (Carter, W. G., Wayner, E. A., Bouchard, T. S., and Kaur, P. (1990) J. Cell Biol. 110, 1387-1404; Kaufmann, R., Frosch, D., Westphal, C., Weber, L., and Klein, C. E. (1989) J. Cell Biol. 109, 1807-1815; Hynes, R. O. (1987) Cell 48, 549-554; Symington, B. E., Takada, Y., and Carter, W. G. (1993) J. Cell Biol. 120, 523-535). We present data herein suggesting that the integrin alpha 3 beta 1 may interact homophilically in such cell-cell adhesion structures which contain this specific receptor or, alternatively, in receptor aggregates found in focal adhesions. The alpha 3 beta 1 receptor was purified by affinity chromatography on either human laminin or peptide GD-6 Sepharose and subsequently used as a substrate in cell adhesion assays. The immobilized alpha 3 beta 1 supported the adhesion of cells containing alpha 3 beta 1, and this attachment was specifically inhibited by monoclonal antibodies to both beta 1 and alpha 3 subunits. In addition, an affinity matrix containing purified alpha 3 beta 1 showed specific binding of only alpha 3 beta 1 from detergent extracts of cell surface proteins and such binding was cation dependent. Finally, using biosensor technology involving the principle of surface plasmon resonance (BIAcore, Pharmacia Biosensor), alpha 3 beta 1, when bound to a carboxymethyl dextran-modified gold surface, was found to bind only other soluble alpha 3 beta 1 receptors and did not bind other purified integrins, including alpha 5 beta 1 and alpha v beta 3. These data strongly suggest that alpha 3 beta 1 likely interacts in a homophilic manner under our experimental conditions. PMID- 8408062 TI - Repair of oxidative damage within the mitochondrial DNA of RINr 38 cells. AB - A growing body of evidence suggests that a variety of chronic diseases, including cancer and diabetes, are associated with damage to mitochondrial DNA. Since mitochondria are constantly exposed to high levels of reactive oxygen species, it is likely that oxidative damage to mitochondrial DNA may be responsible for some of these maladies. To determine whether mitochondria can repair this damage, a quantitative Southern blot technique was utilized to identify repair in specific DNA fragments. A 10.8-kilobase mitochondrial restriction fragment was studied employing a probe containing the entire mouse mitochondrial genome. Alloxan was employed to generate oxygen radicals. Insulinoma cells were exposed to alloxan for 1 h, and total cellular DNA was isolated immediately or after intervals of up to 8 h. Alkali treatment was used to identify abasic sites and sugar lesions, endonuclease III was used to identify lesions associated with thymine and cytosine damage, and formamidopyrimidine-DNA glycosylase was employed to recognize formamidopyrimidines and 8-oxoguanines in DNA. The results showed that all forms of damage studied were repaired by 4 h, indicating that mitochondria are able to efficiently repair damage to their DNA caused by reactive oxygen species. PMID- 8408063 TI - Bombesin, vasopressin, and endothelin rapidly stimulate tyrosine phosphorylation of the focal adhesion-associated protein paxillin in Swiss 3T3 cells. AB - Treatment of Swiss 3T3 cells with bombesin caused a striking increase (21-fold) in the tyrosine phosphorylation of the cytoskeleton-associated protein paxillin, as judged by anti-phosphotyrosine Western blots of anti-paxillin immunoprecipitates. Vasopressin and endothelin also stimulated paxillin tyrosine phosphorylation. Bombesin-stimulated tyrosine phosphorylation of paxillin was detectable within 1 min and was concentration-dependent (half-maximum effect at 0.09 nM). Bombesin stimulation of paxillin tyrosine phosphorylation could be dissociated from both protein kinase C (PKC) activation and the mobilization of Ca2+ from intracellular stores. Activation of PKC in quiescent Swiss 3T3 cells using the tumor promoter phorbol 12,13-dibutyrate (PDB) increased the tyrosine phosphorylation of paxillin in a time-dependent manner but was less effective than bombesin and stimulated detectable phosphorylation only within 5 min, considerably slower than bombesin-induced tyrosine phosphorylation of paxillin. Furthermore, the selective PKC inhibitor, GF109203X, or down-regulation of PKC using prolonged treatment with PDB markedly inhibited the stimulation of paxillin tyrosine phosphorylation by PDB but had little effect on the response to bombesin. In contrast, cytochalasin D, an agent that selectively disrupts the network of actin microfilaments, completely inhibited bombesin- and PDB-induced paxillin tyrosine phosphorylation. This is the first report to identify paxillin as a substrate for neuropeptide-stimulated tyrosine phosphorylation. PMID- 8408064 TI - The carboxyl-terminal domain of the human pregnancy-specific glycoprotein specifies intracellular retention and stability. AB - The pregnancy-specific glycoproteins (PSGs), which are members of the immunoglobulin superfamily, are the major pregnancy-associated proteins synthesized by the human placenta. Thirty or more PSG members have been identified which are encoded by at least 11 linked genes. The PSG proteins share 85-95% sequence homology in the coding region, but show variability at the carboxyl-terminal (COOH) domains. In the present study, we examined the effects of PSG COOH domains on protein secretion and stability. Using PSGs containing short (11-12 residues) hydrophilic (PSG1e, PSG11s, and PSG16a), short (22 residues) hydrophobic (PSG6r), and long (81 residues) hydrophobic (PSG11w) COOH domains, we showed that most PSG members were secretory proteins except PSG11w which was largely retained in cells. When the PSG11w COOH domain was replaced with a short COOH domain of PSG1e, the resulting PSG-N11w/C1e chimera became secreted into the medium. On the other hand, chimeras that harbored the PSG11w COOH domain, PSG-N1e/C11w and PSG-N16a/C11w, remained in cells, demonstrating that the COOH domain of PSG11w confers intracellular retention. Deletion analysis showed that mutant (PSG11w-C2) that contained the first 21 amino acids of PSG11w COOH domain or mutant (PSG11w-C3) that contained a deletion of hydrophobic residues 372-392 in the PSG11w COOH domain remained largely in cells. In contrast, the PSG11w-C1 mutant which contained the first 12 residues of the PSG11w COOH domain became a secretory protein. Studies of PSG synthesis and processing in the presence of Brefeldin A, a drug that impedes protein transport from endoplasmic reticulum to the Golgi system, showed that PSG11w resided and degraded in the endoplasmic reticulum. The endoplasmic reticulum localization of PSG11w and the cell-associated mutant PSGs was further demonstrated by their sensitivity to endoglycosidase H and indirect immunofluorescence analysis. PMID- 8408065 TI - Identification of sites on the human Fc epsilon RI alpha subunit which are involved in binding human and rat IgE. AB - The high affinity IgE Fc receptor (Fc epsilon RI), found on mast cells and basophils, is a tetrameric receptor complex. The extracellular portion of the Fc epsilon RI alpha subunit consists of two immunoglobulin-like domains and binds IgE in the absence of the other subunits. To localize the high affinity IgE binding site within the Fc epsilon RI alpha subunit, we generated a series of chimeric receptor constructs where one of the two immunoglobulin-like domains was either deleted or substituted with those from the human Fc gamma RIIIA alpha or the rat Fc epsilon RI alpha subunit. The chimeric receptors were monitored for their capacity to bind human and rat IgE, and their reactivity with different antireceptor antibodies. Domain I substitutions maintained high affinity human IgE binding. Domain II substitutions resulted in a total loss of both human and rat IgE binding. Single-domain alpha subunits could not bind IgE, suggesting that both extracellular domains are required for proper protein folding or IgE binding. To further localize the IgE binding sites, homolog-scanning mutagenesis was performed. At least three independent regions of domain II encompassing residues 118-129, 136-150, and 148-162 were required for IgE binding. Our results suggest that domain II of the human Fc epsilon RI alpha confers most of the important contributions to the binding of the human IgE Fc molecule, whereas domain I of the rat Fc epsilon RI alpha makes important contributions to the binding of rat IgE. PMID- 8408066 TI - Intracellular retention of interleukin-6 abrogates signaling. AB - Three forms of interleukin-6 (IL-6) have been constructed and stably transfected into human hepatoma cells (HepG2). Wild type IL-6 containing a signal peptide was rapidly secreted as a biologically active protein. IL-6 lacking the signal peptide accumulated within the cytoplasm of transfected cells. Surprisingly, IL-6 carrying a COOH-terminal extension of the amino acids Lys-Asp-Glu-Leu (KDEL) was not completely retained in the endoplasmic reticulum (ER). Complete retention in the ER was achieved when the 14 COOH-terminal amino acids of protein disulfide isomerase which include the KDEL signal were added to the COOH terminus of IL-6. This finding clearly demonstrates that the addition of the protein sorting signal KDEL alone is not sufficient for full retention of IL-6 in the ER. IL-6 accumulated in the cytoplasm and IL-6 retained in the ER failed to induce liver specific acute-phase protein synthesis in the host cells, indicating that there is no intracellular role for IL-6 in signal transduction. Retention of IL-6 in the ER led to the prevention of surface expression of the IL-6 receptor protein gp80, making these cells unresponsive to IL-6. This phenomenon can be exploited in the future to generate transgenic animals which will become completely cytokine unresponsive in the tissues in which they express an ER retained cytokine. PMID- 8408067 TI - Crystal structures of ribonuclease HI active site mutants from Escherichia coli. AB - In order to investigate the relationships between the three-dimensional structure and the enzymic activity of E. coli RNase HI, three mutant proteins, which were completely inactivated by the replacements of three functional residues, Asp10 by Asn (D10N), Glu48 by Gln (E48Q), and Asp70 by Asn (D70N), were crystallized. Their three-dimensional structures were determined by x-ray crystallography. Although the entire backbone structures of these mutants were not affected by the replacements, very localized conformational changes were observed around the Mg(2+)-binding site. The substitution of an amide group for a negatively charged carboxyl group in common induces the formation of new hydrogen bond networks, presumably due to the cancellation of repulsive forces between carboxyl side chains with negative charges. These conformational changes can account for the loss of the enzymic activity in the mutants, and suggest a possible role for Mg2+ in the hydrolysis. Since the 3 replaced acidic residues are completely conserved in sequences of reverse transcriptases from retroviruses, including human immunodeficiency virus, the concepts of the catalytic mechanism deduced from this structural analysis can also be applied to RNase H activity in reverse transcriptases. PMID- 8408068 TI - Identification of a nuclear localization sequence within the structure of the human interleukin-1 alpha precursor. AB - Recent studies have suggested that the signal peptide-less cytokine, interleukin (IL)-1 alpha, may play a role as an intracellular regulator of human endothelial cell proliferation in vitro (Garfinkel, S., Haines, D. S., Brown, S., Wessendorf, J., Gillespie, D. H., and Maciag, T. (1992) J. Biol. Chem. 267, 24375-24378). In order to determine the intracellular locale of the IL-1 alpha precursor, we fused the open reading frame of the IL-1 alpha precursor to the reporter gene beta galactosidase (Gal) and studied the cellular distribution of the chimera in NIH 3T3 cells after transfection. Immunological and enzymatic analysis demonstrated that the IL-1 alpha:beta-Gal fusion protein was associated with the nucleus. To further define the region responsible for this activity, we ligated the mature form of IL-1 alpha (IL-1 alpha 113-271) and the IL-1 alpha precursor domain (IL-1 alpha 1-112) to beta-Gal. Analysis of the intracellular distribution of these chimeric polypeptides following transfection demonstrated a differential distribution of IL-1 alpha 1-112:beta-Gal in the nucleus and IL-1 alpha 113 271:beta-Gal in the cytosol. Because the IL-1 alpha precursor domain contains a sequence that resembles a nuclear translocation signal (KVLKKRRL, residues 79 86), we prepared an IL-1 alpha precursor point mutant in which Lys82 was replaced by Glu. Transfection of NIH 3T3 cells with the IL-1 alpha precursor point mutant (IL-1 alpha 1-271 Glu82:beta-Gal) resulted in a significant reduction in the ability of the IL-1 alpha precursor to associate with the nucleus and similar data were obtained as a result of Lys82 mutagenesis in the IL-1 alpha precursor domain (IL-1 alpha 1-112 Glu82:beta-Gal). These data suggest that the IL-1 alpha precursor contains a functional nuclear localization sequence within the structure of the precursor domain and Lys82 is critical for its function. PMID- 8408069 TI - Identification of regions in interleukin-1 alpha important for activity. AB - Saturation mutagenesis of the mature human interleukin-1 alpha (IL-1 alpha) gene has been performed. Following expression in Escherichia coli, the biological and receptor binding activities of the mutant proteins were examined. Most of the molecule could be altered with little effect on either function. More than 3,500 mutants were examined, and only 23 unique amino acid sequences were identified which resulted in an altered ratio of biological to binding activity when compared with wild-type IL-1 alpha. These proteins possessed mutations at 38 of the 159 amino acid residues in IL-1 alpha. Random mutagenesis at several of these positions identified further substitutions that affected activity. Examination of a model for IL-1 alpha localized most of the residues which altered activity along one face of the molecule. This region appears to be distinct from areas of IL-1 which have been postulated to make contact with IL-1 receptor. PMID- 8408070 TI - Reciprocal effects of apolipoprotein and lytic peptide analogs on membranes. Cross-sectional molecular shapes of amphipathic alpha helixes control membrane stability. AB - Apolipoprotein (class A) amphipathic helixes are postulated to act as detergents by virtue of their cross-section being wedge-shaped. Using computer analysis of naturally occurring class A and lytic (class L) amphipathic helixes, we designed two archetypical model peptides. Analogs of these two peptides, incorporating substitutions or modifications of interfacial or basic residues, had the following effects. Class A peptides stabilized bilayer structure, reduced leakage from large unilamellar vesicles and erythrocytes, and inhibited lysis induced by class L peptides. Class L peptides destabilized bilayer structure in model membranes and increased binding of class A peptides to erythrocytes. The ability of class L analogs to lyse membranes and induce inverted lipid phases was reduced by either decreasing the bulk of an interfacial residue, increasing the angle subtended by the polar face, or increasing the bulk of the basic residues. The ability of the class A analog to stabilize bilayer structure and inhibit erythrocyte lysis by class L peptides was enhanced by methylating the Lys residues. These results can be explained by a model that we term the reciprocal wedge hypothesis. By analogy to the reciprocal effects of phospholipid shapes on membrane structure, we propose that the wedge shape of class A helixes stabilizes membrane bilayers, whereas the inverted wedge shape of class L helixes destabilizes membrane bilayers, and, thus, one class will neutralize the effect of the other class on membranes. PMID- 8408071 TI - Glucose transport in L6 myoblasts overexpressing GLUT1 and GLUT4. AB - The roles of the glucose transporter isoforms, GLUT1 and GLUT4, in mediating insulin-stimulated glucose transport were investigated by stably overexpressing the transporters in L6 myoblasts. Levels of GLUT1 and GLUT4 in myoblasts from the cell lines having the highest content of these transporters were approximately 16 and 30-fold higher, respectively, than levels in nontransfected cells. The basal rate of 2-deoxy[3H]glucose uptake was severalfold higher in cells overexpressing GLUT1 than in the parent L6 myoblasts or in control cell lines that were generated by transfecting cells with expression vectors lacking transporter insert. The basal rate was not elevated in any of the lines expressing GLUT4. The net increase in 2-deoxy[3H]glucose uptake produced by insulin was larger in both the GLUT1 and GLUT4 cells than in the control cells. Insulin increased uptake in GLUT4 cells by as much as 6-fold; whereas, the fold increase over basal uptake produced by insulin in GLUT1 cells was comparable to that (2-fold) observed in the control myocytes. Thus, both GLUT1 and GLUT4 can mediate insulin-stimulated glucose transport in L6 myoblasts, although GLUT4 is needed to observe large percentage increases comparable to those observed in skeletal muscle fibers in vivo. In contrast to insulin, the protein phosphatase inhibitors, okadaic acid and calyculin A, inhibited glucose transport in cells expressing either GLUT1 or GLUT4. Calyculin A, which produced a half-maximum effect at 10 nM, was approximately 100 times more potent than okadaic acid in decreasing both basal and insulin-stimulated 2-deoxyglucose uptake. Inhibition of uptake by calyculin A was associated with a decrease in the cell surface concentration of both GLUT1 and GLUT4. These results indicate that increased protein phosphorylation can lead to inhibition of transport mediated by both GLUT1 and GLUT4. PMID- 8408072 TI - Enzymic cleavage as a probe of the molecular structures of mammalian equilibrative nucleoside transporters. AB - We have used enzymic cleavage by trypsin in conjunction with glycosidase digestion to probe the transmembrane topologies and molecular structures of mammalian equilibrative, nitrobenzylthioinosine (NBMPR)-sensitive, nucleoside transport systems. Transporters from four species (human, pig, guinea pig, and rat) and three tissues (erythrocyte, liver, and lung), which differ from each other in size and in their sensitivity to inhibition by the vasodilator dipyridamole, were investigated. Broadly equivalent sites of [3H]NBMPR photolabeling, carbohydrate attachment, and trypsin cleavage were observed for all systems. Results from these experiments demonstrate that molecular weight differences between rat transporters and those from two other species (human and guinea pig) are due largely to oligosaccharide heterogeneity and that the low dipyridamole sensitivity of rat nucleoside transporters is probably a consequence of relatively minor differences in molecular structure. In marked contrast, carbohydrate removal increases the molecular weight difference between the pig erythrocyte transporter and, for example, that in human erythrocytes. This polypeptide difference is limited largely, if not completely, to one tryptic fragment of the protein. In the case of the human erythrocyte transporter, the site of N-linked glycosylation has been located very close to one end of the protein, and the site of NBMPR photolabeling to within 16 kDa of that site. Trypsin cleavage occurs endofacially. Our results provide evidence of substantial structural conservation among mammalian NBMPR-sensitive nucleoside transporters. PMID- 8408073 TI - Functional expression and purification of a homomeric human alpha 1 glycine receptor in baculovirus-infected insect cells. AB - The human alpha 1 glycine receptor (GlyR) was expressed in Sf9 insect cells infected with a recombinant Autographa californica nuclear polyhedrosis baculovirus. Previous studies had indicated that transient expression of this subunit in Xenopus oocytes or human kidney cell lines is sufficient to form active agonist-gated chloride channels. Expression of the alpha 1 GlyR protein resulted in functional channels present on the cell surface of infected Sf9 cells as evidenced by whole-cell patch-clamping and single-channel recordings. These channels were gated by glycine, but not in the presence of strychnine. An immunoreactive 48-kDa protein could be easily visualized on Coomassie-stained sodium dodecyl sulfate-polyacrylamide gels of whole-cell lysates with maximal expression 3 days postinfection. The alpha 1 GlyR protein was solubilized from a membrane fraction of infected Sf9 cells in 1% digitonin and 0.1% deoxycholate and purified by affinity chromatography using aminostrychnine agarose, yielding 0.33 mg/liter of cells. Given the low natural abundance of the native channel, the development of this expression system now provides sufficient purified channel protein for future biochemical and biophysical characterization. Since the glycine receptor shares sequence and structural homology with other members of a ligand-gated channel superfamily, further characterization may establish general rules governing the structure and mechanism of these membrane protein channels. PMID- 8408074 TI - Identification of an inducible 85-kDa nuclear protein kinase. AB - To identify inducible protein kinases localized exclusively in the nucleus, nuclear and cytosolic extracts were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and electroblotted to an Immobilon-P membranes were incubated in phosphorylation buffer containing [gamma-32P]ATP. Autoradiographs of the membranes revealed an 85-kDa 32P-labeled band; the intensity of this band was transiently increased in nuclear but not in cytosolic extracts from interleukin-1 alpha-treated cells. Incorporation of 32P label into a blotted protein band suggested the presence of an interleukin-1 alpha responsive 85-kDa nuclear protein kinase. Fractionation of nuclear extracts by Mono Q failed to separate the kinase activity from the substrate, indicating that the 85-kDa band identified on the Immobilon-P membrane represents a protein kinase that undergoes autophosphorylation. Phosphoamino acid analysis of the 85 kDa band showed that this enzyme is a serine/threonine kinase. Purified pp90RSK could not be identified by the denaturation-renaturation method, indicating that the 85-kDa kinase identified here is not pp90RSK. This observation, nuclear but not cytoplasmic localization, and the fact that antibodies to known protein kinases kinase failed to recognize it suggest that the enzyme identified here is a novel protein kinase. PMID- 8408075 TI - Detection and characterization of ceramide-1-phosphate phosphatase activity in rat liver plasma membrane. AB - A calcium-dependent ceramide (Cer) kinase was recently detected in human leukemia (HL-60) cells (Kolesnick, R.N., and Hemer, M.R. (1990) J. Biol. Chem. 265, 18803 18808) where it may function in terminating the regulatory effects of Cer, and in synaptic vesicles (Bajjalieh, S. M., Martin, T. F. J., and Floor, E. (1989) J. Biol. Chem. 264, 14354-14360). We now demonstrate that the addition of both Cer-1 phosphate (Cer-1-P) and a short-acyl chain analog of Cer-1-P,N hexanoylsphingosine-1-phosphate (C6-Cer-1-P) to cultured cells and a variety of subcellular fractions results in rapid degradation to Cer and C6-Cer, respectively. The Cer-1-P phosphatase activity is enriched in a rat liver plasma membrane fraction and appears to be distinct from the phosphatase that hydrolyzes phosphatidic acid (PA), PA phosphohydrolase, as shown by the difference in sensitivity of Cer-1-P and PA hydrolysis to propranolol, detergent, and heat treatment. Moreover, the Km of Cer-1-P hydrolysis is 10-fold lower than the Km of PA hydrolysis in plasma membrane. PA is a noncompetitive inhibitor of Cer-1-P hydrolysis, with an inhibition constant 1-1.5-fold higher than the Km of Cer-1-P hydrolysis. In contrast, Cer-1-P does not inhibit PA hydrolysis. Finally, we describe the synthesis of a novel analog of Cer-1-P which is not hydrolyzed in vitro and in vivo and is internalized in cultured cells by endocytosis. These results are discussed in relation to the possible roles of Cer-1-P in regulating intracellular levels of Cer. PMID- 8408076 TI - A suppressor gene that enables Saccharomyces cerevisiae to grow without making sphingolipids encodes a protein that resembles an Escherichia coli fatty acyltransferase. AB - Saccharomyces cerevisiae normally requires sphingolipid biosynthesis for growth; however, mutant strains lacking sphingolipids have been isolated by suppression of a genetic defect in sphingolipid long chain base biosynthesis. To begin to understand the nature of the suppressor(s) we isolated and characterized a suppressor gene, SLC1 (sphingolipid compensation). DNA sequence analysis showed that the wild type SLC1 allele differs from the suppressor allele by a single nucleotide which changes Gln-44 in the predicted wild type protein to Leu4-4 in the predicted SLC1-1 suppressor protein. The predicted SLC1 protein sequence is homologous to the 1-acyl-sn-glycerol-3-phosphate acyltransferase of Escherichia coli encoded by the plsC gene. The homology extends to function as well since the SLC1 gene complements the growth defect in an E. coli strain mutated in plsC. These results suggest that the SLC1 protein has a fatty acyltransferase activity. SLC1 thus may be the first eucaryotic sn2-acylglyceride fatty acyltransferase gene to be cloned. SLC strains grown in the absence of long chain base make novel phosphatidylinositol derivatives (Lester, R. L., Wells, G. B., Oxford, G., and Dickson, R. C. (1993) J. Biol. Chem. 268, 845-856) having a C26 fatty acid at the sn-2 position and the same polar head groups as normal sphingolipids. We postulate that the SLC1 suppressor allele encodes a variant enzyme with an altered substrate specificity that enables it to use a C26 in place of a C16/18 fatty acid precursor to acylate the sn-2 position of inositol-containing glycerolipids. PMID- 8408077 TI - cis-acting elements in the 5'-untranslated region of rat testis proenkephalin mRNA regulate translation of the precursor protein. AB - We found that a rat testis cDNA library contained two types of proenkephalin cDNA, corresponding to somatic and germ cell-specific transcripts. The somatic form is identical in primary sequence to previously characterized transcripts from rat brain and heart. The longer germ cell-specific cDNA contains a novel 5' terminus derived from the somatic intron A. We found that germ cell-specific transcripts translate in vitro with significantly lower efficiency than do somatic-type transcripts. Deletion analysis indicated that translational inefficiency is mediated primarily by the presence of four upstream AUG codons followed by short open reading frames within the leader sequence. Secondary structure in the form of a stem-and-loop upstream of the initiator codon for proenkephalin was found to have a slightly inhibitory effect on translation. These results, coupled with the very low abundance of the somatic transcript, provide a possible explanation for the low levels of opioid peptides found in adult rat testis in spite of the abundant quantity of germ cell-specific proenkephalin transcript. PMID- 8408078 TI - Characterization of the structural requirements and cell type specificity of IL-1 alpha and IL-1 beta secretion. AB - The murine macrophage cell line, P388D1, was transfected with expression plasmids containing a cDNA for the precursor or mature form of human interleukin (IL)-1 alpha or -1 beta, and the extent of IL-1 protein secretion was analyzed. The secretion of IL-1 beta from cells transfected with a cDNA encoding the mature IL 1 beta was approximately 15-fold greater than from cells transfected with a precursor IL-1 beta cDNA. Although P388D1 cells transfected with precursor and mature IL-1 alpha cDNAs exhibited similar levels of secretion, the precursor IL-1 alpha cDNA-transfected cells secreted only the mature form of IL-1 alpha. Relatively high levels of IL-1 secretion were also observed in cultures of HeLa cells and the murine EL 4 and human Jurkat T cell lines that were transfected with a mature IL-1 alpha or -beta expression plasmid. Secretion was not observed when these cell lines were transfected with precursor IL-1 alpha or -beta expression plasmids. Secretion was also minimal when P388D1 cells were transfected with cDNAs encoding mature IL-1 beta proteins lacking the NH2- or COOH-terminal 20 amino acids. These results indicate that the mature forms of IL 1 proteins are the preferred substrates for secretion and that the maintenance of a specific conformation of these proteins may be required for optimal secretion. PMID- 8408079 TI - Characterization of a chloroplast homologue of the 54-kDa subunit of the signal recognition particle. AB - Proteins and RNAs homologous to components of the eukaryotic signal recognition particle (SRP) have been identified in a number of prokaryotic organisms. Here we report the isolation of an Arabidopsis thaliana cDNA, FFC (fifty-four chloroplast homologue), that encodes a chloroplast protein (54CP) homologous to the 54-kDa subunit of the signal recognition particle. 54CP shares 27% identity with mammalian SRP54 and 44% identity with the Escherichia coli ffh gene product suggesting a prokaryotic origin for this gene. FFC is a nuclear gene encoding a 62-kDa cytoplasmically synthesized precursor that is capable of being imported into isolated chloroplasts and processed to a 53-kDa stromal polypeptide. Antibodies generated against a portion of 54CP recognize an endogenous 53-kDa chloroplast protein that sediments at 4 S and 70 S, the latter form may be associated with ribosomes. The FFC transcript is most abundant in green shoot tissue whereas etiolated buds and roots have transcript levels about 30 and 10%, respectively, relative to light grown green shoots. By analogy with the eukaryotic signal recognition particle which targets secretory proteins to the endoplasmic reticulum, it is speculated that 54CP may target chloroplast proteins to either the thylakoid or envelope membranes. PMID- 8408080 TI - Unfolding of newly made retinol-binding protein by dithiothreitol. Sensitivity to retinoids. AB - In cultured hepatoma HepG2 cells, serum retinol-binding protein (RBP) is secreted more rapidly in the presence of retinol than in its absence (Tosetti, F., Ferrari, N., Pfeffer, U., Brigati, C., and Vidali, G. (1992) Exp. Cell Res. 200, 467-472). In the presence of millimolar concentration of DTT, HepG2 cells synthesize fully reduced RBP within the endoplasmic reticulum (ER) which, upon removal of DTT, forms disulfide bonds post-translationally. Secretion of this post-translationally folded RBP is also dependent on the presence of retinol. Using nonreducing gel electrophoresis, we resolved disulfide-bonded RBP folding intermediates. In addition, two other intracellular folding intermediates, compact I and II, which co-migrate with mature RBP were resolved by their different sensitivity to DTT-induced unfolding. Retinol, as well as retinoic acid, stabilized both compact I and II RBP intermediates to DTT-induced unfolding, suggesting that RBP assumes different conformations in the ER in the presence and absence of a ligand. However, only RBP synthesized in the presence of retinol is rapidly secreted, indicating that the ER export quality control system recognizes RBP containing retinol, but not retinoic acid, as fully folded and competent for export. Folding of RBP so that it is stabilized to DTT reduction is not a sufficient condition for ER exit. PMID- 8408081 TI - In vitro unfolding of retinol-binding protein by dithiothreitol. Endoplasmic reticulum-associated factors. AB - HepG2 cells in the presence of DTT synthesize fully reduced serum retinol-binding protein (RBP) within the endoplasmic reticulum (ER). Upon removal of DTT, RBP forms disulfide bonds and a folding intermediate, compact II, accumulates within the ER. Compact II RBP co-migrates on nonreducing gel electrophoresis with the secreted form of RBP but is differentiated from secreted RBP by its sensitivity to DTT-induced unfolding (see accompanying article; Kaji, E. H., and Lodish, H. F. (1993) J. Biol. Chem. 268, 22188-22194). Here, we have reconstituted DTT induced unfolding of compact II RBP in a broken cell system and demonstrate that ER-associated factors enhance the unfolding of RBP by DTT. Protein disulfide isomerase is likely to be one such factor since it enhances the rate of RBP unfolding by DTT in vitro; protein disulfide isomerase-induced unfolding requires the absence of retinoids, similar to the DTT-induced unfolding in vivo. ATP enhances the unfolding of RBP in the absence but not in the presence of retinol, both in intact and broken cells. Thus, protein disulfide isomerase and other ATP dependent factors can unfold partly folded (or misfolded) RBP in the ER, suggesting how improperly folded proteins might be correctly refolded in vivo. PMID- 8408082 TI - Molecular characterization of the murine Ahr gene. Organization, promoter analysis, and chromosomal assignment. AB - The AH receptor is a ligand-activated transcription factor that mediates the biological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin. The AH receptor has primary sequence homology to its dimerization partner the AH receptor nuclear translocator, and to the Drosophila proteins Sim and Per. Characterization of the gene encoding the murine AH receptor (Ahr gene) reveals that its structural organization is also conserved with respect to the sim gene, since 6 of 11 Ahr exons are spliced at homologous sites. Interestingly, little splicing homology was observed between the Ahr and per genes. The promoter of the Ahr gene is GC rich and contains no TATA or CCAAT boxes; however, sequence analysis has shown several binding sites for the transcription factor Sp1 (GC boxes). Additionally we have identified a potential cAMP response element, AP-1 and E box sites, and two elements demonstrated in other genes to confer placenta-specific expression. Using a restriction fragment length polymorphism in exon 7 and recombinant inbred mouse lines, the Ahr gene was found to be concordant with the phenotypically defined Ahr locus, supporting the identity of these two genetic elements. PMID- 8408083 TI - The subunit location of magnesium in cytochrome c oxidase. AB - The magnesium ion in bovine heart cytochrome c oxidase can be depleted up to 75% by heat treatment of the enzyme at 43 degrees C followed by dialysis against EDTA buffer solution. The magnesium-depleted enzyme so obtained retains 40% of the activity of the native enzyme. This is the first attempt to deplete magnesium ion from bovine heart cytochrome c oxidase without denaturation of the protein. Magnesium depletion exposes at least one carboxyl group on subunit IV for labeling by N-cyclohexyl-N'-(4-dimethylaminonaphthyl)carbodiimide (NCD-4). The NCD-4 labeling of subunit IV of the magnesium-depleted enzyme is significantly enhanced relative to what is observed for the native and heat-treated oxidase, suggesting that the magnesium ion is located in subunit IV with at least one carboxyl ligand. By comparing the activity of the magnesium-depleted enzyme with that of a control sample of heat-treated oxidase, the influence of divalent magnesium on the activity of the enzyme is assessed. PMID- 8408084 TI - Analysis of biomechanical and ergonomic aspects of the cervical spine in F-16 flight situations. AB - With the help of a biomechanical neck model, several normal postures of an F-16 pilot were analysed. Measurements of accelerations and head positions were obtained during four flights, including simulated air combat. With the help of a model, muscle forces and joint reaction forces in the neck were estimated. Although at the present stage of research results of calculations must be interpreted carefully, conclusions can be drawn with respect to sitting posture, head position and helmet devices. The backward inclined back rest of the F-16 chair decrease the lordosis of the cervical spine, resulting in reduced calculated forces in the lower cervical spine. In high load situations, calculated maximal forces are of the same order of magnitude as failure loads of vertebrae and estimations of maximum muscle forces. The calculated neck load is increased substantially by the helmet and helmet-mounted devices. This load can be reduced by lightening the helmet or shifting the centre of mass of the helmet backwards. PMID- 8408085 TI - The role of fixator frame stiffness in the control of fracture healing. An experimental study. AB - External skeletal fixation is used widely in the management of fractures. Frame configuration is known to affect frame stiffness and, thereby, the local mechanical environment at the fracture site. In previous investigations of the influence of mechanical conditions upon fracture healing, the frames have always been applied so that they influence the biological environment in different ways. As a result, the influence of stiffness, per se, could not be studied as a single variable, and its effect on the repair process remains unclear. In this study, using a standard osteotomy of the ovine tibia, stabilised by an external skeletal fixator, the local mechanical environment was altered solely by increasing the 'offset' distance between the bone and the fixator frame. The biological conditions at the fracture remained identical in both groups. Increasing the frame stiffness by 40%, brought about by reducing the offset distance of the fixator bar by 10 mm, caused a significant reduction in the rate of healing. In addition, the frame stiffness influenced the ground reaction force with greater weight-bearing in the initial stages in the more rigid group, but despite this, the resultant interfragmentary displacement in this group appeared to be insufficient to stimulate fracture healing. This work emphasises the importance of the local mechanical environment on the process of fracture healing. It also demonstrates the value of in vivo assessment of fracture stiffness as a means of monitoring mechanical events during fracture healing. PMID- 8408086 TI - Kinetics of the lower extremities during drop landings from three heights. AB - In this study, the landing preferences of gymnasts (n = 6) and recreational athletes (n = 6) were determined by comparing the changes in lower extremity kinetics of drop landings performed from three heights (0.32-1.28 m). Net joint moments and work done on the extensor muscles of the ankle, knee, and hip were selected as variables representative of the demand placed on the muscles responsible for controlling flexion and dissipating the load. Kinematic and kinetic two-dimensional data were acquired simultaneously using high-speed film (202.4 fps) and a force plate (1000 Hz). Reaction forces and lower extremity joint motions were used to calculate net joint forces, net joint moments powers, and work done on the extensor muscles of the ankle, knee, and hip. Results indicated that the extensor joint moments tended to peak earlier after contact with increases in velocity, but the temporal sequence of events was maintained independently of velocity or group. As impact velocity increased, net peak extensor moments and work done on the extensor muscles significantly increased. Significantly larger ankle and hip peak extensor moments were observed for the gymnasts across velocities as compared to the recreational athletes. No significant differences in work done on the extensor muscles were noted between groups. Significant interaction effects indicate that gymnasts chose to dissipate the loads at contact by using larger ankle and hip extensor moments at higher impact velocities than the recreational athletes, whereas recreational athletes chose to adjust their strategy by using greater degrees of hip flexion (McNitt Gray, Int. J. Sport Biomech, 7, 201-204, 1991) and longer landing phase durations than the gymnasts. The greater demands placed on the ankle and hip extensors by the gymnasts, as compared to the recreational athletes, may be explained by the need to maintain balance during competitive gymnastics landings or, perhaps, by the inability of recreational athletes to produce larger extensor moments at the ankle or hip during landings from great heights. PMID- 8408087 TI - The effects of collagen fiber orientation, porosity, density, and mineralization on bovine cortical bone bending properties. AB - The relationship between the mechanical properties of bone in three-point bending and eight histocompositional variables was studied. Ultimate stress, ultimate strain, and elastic modulus were measured in 35 beams of cortical bone from bovine tibias using standard ASTM methods. Four elements of porosity were determined by point counting, mineralization by ashing, and wet and dry apparent density from weight and volume. Collagen fiber orientation was estimated using polarized light, and specimens were categorized as plexiform, mixed, or osteonal. Analysis of variance showed that ultimate stress was similar in the plexiform and osteonal specimens, but elastic modulus was reduced in the latter (18.6 +/- 1.2 vs 21.0 +/- 1.9 GPa), which were significantly less porous (by 24%) and less mineralized (by 3%) than the plexiform group. Stepwise multiple regression analysis showed that collagen fiber orientation ranked highly as a predictor of bending properties. The next best predictors were density and mineralization. In the plexiform group, 77% of the variability in elastic modulus was accounted for by wet and dry density and collagen fiber orientation. In the osteonal group, 88% of modulus variability was accounted for by percentage mineralization and collagen fiber orientation. When all the specimens were pooled, 62% of the variability in elastic modulus was attributable to variations in collagen fiber orientation, density, and porosity due to Haversian canals. PMID- 8408088 TI - The effects of dehydration and rehydration on some mechanical properties of human dentine. AB - This study was designed to investigate the effect of dehydration and rehydration on the brittleness and toughness of human dentine. Tensile and three-point bend tests were carried out on hydrated, dehydrated and rehydrated dentine bars, sectioned from sound extracted, human third molar teeth. The stress, strain and fracture energy (toughness) were calculated and the results were analysed using ANOVA and Duncan's multiple range test at p = 0.01. Stress at fracture did not differ significantly between hydrated, dehydrated or rehydrated dentine in bending or tensile tests. Strain at fracture and fracture energy were significantly greater for hydrated and rehydrated than for dehydrated dentine. In bending, the elastic energy (resilience) of dehydrated dentine was significantly greater than that of hydrated or rehydrated dentine, but dehydrated dentine showed no plastic energy (deformation) in contrast with the high values for hydrated and rehydrated dentine. Dehydration of human dentine resulted in decreased strain at fracture and demonstrated a brittle behaviour. The absence of plastic energy of deformation and the significantly reduced energy required to induce fracture were indicative of decreased toughness by dehydration. These changes were abolished after rehydration. PMID- 8408089 TI - Joint moments and muscle activity in the lower extremities and lower back in lifting and lowering tasks. AB - The mechanical loading on the body during the act of lifting has been estimated frequently. The opposite act of lowering has received much less attention. The aim of the present study was to compare the mechanical loading of the musculoskeletal system in lifting and lowering. Eight subjects repetitively lifted and lowered a load, using two different techniques (a leg and a back technique). The ankle, knee, hip and lumbosacral joint moments were estimated and the myoelectrical (EMG) activity of seven (leg and back) muscles was recorded. The differences between the lifting and lowering phase for the leg technique were similar to those observed when the back technique was applied. The joint moment curves in lifting showed a high level of agreement with the (time reversed) moment curves in lowering. Peak moments in lowering were only slightly lower than in lifting (peak lumbar moments were 5.4% lower). These small differences were related to different acceleration profiles at the centre of gravity of the body/load complex. The EMG activity was considerably lower in lowering than in lifting. The mean EMG in lowering (average for seven muscles) was only about 69% of the EMG in lifting. This was attributed to the different types of muscle actions involved in lifting (mainly concentric) and lowering (mainly eccentric). Furthermore, the EMG results suggest that similar inter muscular coordination is involved in lowering and lifting. The results give rise to the assumption that in lifting and lowering similar muscle forces are produced to meet the (nearly) equal joint moments, but in lowering these forces are distributed over a smaller cross-sectional area of active muscle, which might imply a higher risk of injury. PMID- 8408090 TI - Chaos in the discrete-time algorithm for bone-density remodeling rate equations. AB - We compare the predictions of the differential equation form of a class of bone density stress adaptation models with their associated discrete-time computational algorithms. Although our considerations apply to the class of adaptation models based on bulk or apparent bone-density remodeling, we focus attention on a particular model in this class, a model employed by Weinans et al. [Trans. Orthop. Res. Soc. 14, 310 (1989); Trans. First World Congress of Biomechanics, Vol. II, p. 75 (1990)]. We show that the discrete-time computational algorithm of that stress adaptation model has a well-known chaos mechanism for stress values of practical interest. Further, we obtain a condition on the discrete-time step that prevents the transition to chaos, and conditions that insure monotonic convergence. This chaos mechanism is only present in the discrete-time computational algorithm; we show that the corresponding differential equation form of the bone-density stress adaptation model is smooth, monotonic and nonchaotic. PMID- 8408091 TI - Biomechanical model of the human foot: kinematics and kinetics during the stance phase of walking. AB - A model of the human foot is proposed in which the foot is represented as eight rigid segments and eight monocentric, single-degree-of-freedom joints. The soft tissue under the foot is divided into seven independent sites of contact, or loading, and each of these is modelled as a nonlinear spring and a nonlinear damper in-parallel. The model was used to estimate the kinematics and kinetics of the foot during the stance phase of walking. The force sustained at each loading site was calculated from walking trials in which only portions of the foot landed on a small force platform. The position of the calcaneus was defined by surface markers, whereas the position of the distal segments were based upon chalk footprints and an estimate of the compression of the plantar soft tissue. The results suggest that the joints that constitute the longitudinal arch extend slightly when the forefoot is loaded. During push-off, these joints flex as the metatarsophalangeal joints extend. Similar kinematic results were estimated when the distal segments of the foot were defined by surface markers. The magnitude of the joint moments of force depended largely on the distribution of the load under the foot which varied considerably between subjects. The stable, yet resilient properties of the foot, as highlighted by this model, should be considered in three-dimensional dynamic models used to study human locomotion. The model provides an objective tool to quantify foot motion and loading, which may prove useful for describing foot function in normal and pathological conditions. PMID- 8408092 TI - Flow disturbances in early femoral atherosclerosis--an in vivo study with digitized cineangiography. AB - Disturbances in arterial flow are believed to influence the localization and development of atherosclerotic plaques. The femoral arteries of 26 patients were studied with cineangiography, after which the films were digitized and analyzed with an image analysis computer. The image sequence was converted to a set of time-intensity curves, from which time parameters were calculated, representing the arrival time of the contrast medium at each pixel. In the resulting parametric images, zones of delayed contrast filling (ZDF) were identified by an adaptive thresholding, which identifies lighter regions within the vessel, excluding the smallest ones. The ZDF, which have been shown, in a model study, to correspond to disturbed flow, were more frequent in the inner curvature than in the outer curvature of the curved vessels. Accordingly, they were more frequent along the lateral wall than along the medial wall of the artery. Several ZDF were also found in the vicinity of bifurcations. Most of the findings tally closely with fluid mechanical theory. In future studies, flow disturbances are to be correlated with the progression of atherosclerotic lesions. PMID- 8408093 TI - Muscle force as affected by fatigue: mathematical model and experimental verification. AB - The effects of muscle fatigue have been included in a comprehensive muscle model derivation. The fiber-based muscle model calculates muscle force as the sum of individual fiber forces, determined based on fiber kinematics and activation information. Three fiber types are considered in the model derivation. Fiber force is modified using indices of fatigue which were derived based on the data from experimental work conducted by others. In this paper, derivations of the indices for each of the fiber types are presented. An elbow extension study was performed to validate the modeled fatigue relationship for maximal effort activities. Triceps brachii forces generated during both isometric and dynamic fatigue tests were determined using both the muscle model and a direct approach based on rigid-body dynamics. The forces obtained from the two approaches were not significantly different for times either less than 50 s for the isometric tests, or greater than 40 s for the dynamic tests. Results from the statistical comparison indicate the utility of the modeling approach presented. PMID- 8408094 TI - A non-Newtonian fluid model for blood flow through arteries under stenotic conditions. AB - This paper presents an analytical study on the behaviour of blood flow through an arterial segment having a mild stenosis. The artery has been treated as a thin walled initially stressed orthotropic non-linear viscoelastic cylindrical tube filled with a non-Newtonian fluid representing blood. The analysis is restricted to propagation of small-amplitude harmonic waves, generated due to blood flow whose wave length is large compared to the radius of the arterial segment. For the equations of motion of the arterial wall consideration is made of a pair of appropriate equations derived by using suitable constitutive relations and the principle of superimposition of a small additional deformation on a state of known finite deformation. It has been shown through numerical computations of the resulting analytical expressions that the resistance to flow and the wall shear increase as the size of the stenosis increases. A quantitative analysis is also made for the frequency variation of the flow rate at different locations of the artery, as well as of the phase velocities and transmission per wavelength. PMID- 8408095 TI - ICI 169,369, A 5-HT2/5-HT1C antagonist, that can evoke endothelium-dependent relaxation in rabbit aorta. AB - 1. The direct effects of ICI 169,369 on vascular reactivity were investigated in rabbit aortic rings with and without endothelium. 2. ICI 169,369 evoked an endothelium-dependent relaxation in aortic rings precontracted with PGF2 alpha. No direct effects on vascular reactivity were found in endothelial denuded rings. 3. The relaxations induced by ICI 169,369 were inhibited by haemoglobin, an agent known to interfere with the responses to endothelium-derived nitric oxide (EDRF) but not by the cyclo-oxygenase inhibitor, indomethacin. Inhibition of the ICI 169,369-induced relaxation by the L-arginine analogue, NG-monomethyl L-arginine (L-NMMA) confirmed that the relaxations evoked by ICI 169,369 were mediated by the endothelial L-arginine: nitric oxide pathway. 4. Studies with competitive receptor antagonists showed that in the rabbit aorta, ICI 169,369 evoked relaxations which were not elicited by the activation of any known 5-HT1, 5-HT2, 5-HT3, muscarinic, histamine, adenosine receptor or adrenoceptors. 5. The diacylglycerol kinase inhibitor, R 59022 also failed to affect these relaxations. It is concluded that ICI 169,369 has a post-receptor action, possibly by directly affecting intracellular calcium levels in the endothelial cells. PMID- 8408096 TI - Inhibition by omega-conotoxin GVIA of the chronotropic responses to sympathetic and parasympathetic nerve stimulation in the isolated, blood-perfused atrium of the dog. AB - 1. We investigated the effects of omega-conotoxin GVIA (omega-CgTX), a blocker of N-type voltage-operated calcium channels, on the chronotropic response to stimulation of the intracardiac sympathetic and parasympathetic nerves in the isolated, blood-perfused right atrium of the dog. 2. omega-CgTX (0.3-3 nmol) itself did not affect the sinus rate significantly, but it inhibited the negative followed by positive chronotropic response to simultaneous stimulation of sympathetic and parasympathetic nerves in a dose-dependent manner. 3. omega-CgTX at higher doses (1-3 nmol) inhibited the positive response to sympathetic stimulation more strongly than the negative response to parasympathetic stimulation. omega-CgTX (3 nmol) abolished the positive chronotropic response to sympathetic nerve stimulation in the atrium treated with atropine, but did not abolish the negative response to selective parasympathetic stimulation. Neither the chronotropic response to noradrenaline nor the response to acetylcholine was affected by omega-CgTX. 4. These results indicate that omega-CgTX inhibits not only the response to sympathetic stimulation but also the response to parasympathetic stimulation in the dog heart and it inhibits the positive chronotropic response to sympathetic stimulation more strongly than the negative chronotropic response to parasympathetic stimulation. PMID- 8408097 TI - Pertussis toxin sensitive effects of dipyridamole on rat atrial rate. AB - 1. In the spontaneously beating rat isolated atria the effects of dipyridamole (0.01, 0.1, 1 and 10 microM) and of adenosine (10 microM) on the chronotropic responses to exogenous noradrenaline (NA) were compared. 2. Dipyridamole (0.01 microM) reduced the chronotropic responses to NA throughout the entire concentration-response curve. A decrease in the maximal response to the agonist was also observed. 3. Neither the spontaneous outflow of [3H]-NA nor its metabolic distribution were altered by dipyridamole (0.01 microM). 4. As observed with dipyridamole, the concentration-response curve to NA was shifted to the right by 10 microM adenosine. 8-Phenyltheophylline (8-PT), 10 microM prevented the decrease in the chronotropic response to NA produced by both 10 microM adenosine and 0.01 microM dipyridamole. 5. The preincubation of rat atria with 1 micrograms ml-1 pertussis toxin prevented the diminution in the chronotropic responses to NA produced by 0.01 microM dipyridamole. 6. The present results suggest that the decrease caused by dipyridamole in rat atrial chronotropic responses involves the participation of adenosine, probably through the interaction with type A1 adenosine receptors. PMID- 8408098 TI - Neurotoxicity and induction of fibroblast proliferation by acetaldehyde in the rabbit liver. AB - 1. Acetaldehyde may, directly or through the formation of condensation products with biogenic amines, be involved in the pathogeny of alcoholic disease. 2. Rabbits were treated acutely (200 mg kg-1, i.p., 1 h before sacrifice) or subacutely (200 mg kg-1 per day, during 5 days; sacrificed 2 days after end of treatment) with acetaldehyde. Another group was administered 6-OHDA (2 x 50 mg kg 1 on day 0 and on day 1, killed on day 5). 3. Acetaldehyde induced a depletion of hepatic noradrenaline. Both in the acute experiments and 2 days after the subacute treatment with acetaldehyde the levels of hepatic noradrenaline were 25% of control. These effects were similar to, but less intense than those induced by 6-OHDA. 4. Both subacute acetaldehyde and 6-OHDA led to a significant increase in the density of fibroblasts in the portal tract spaces of the rabbit liver. 5. The neurotoxic effects of acetaldehyde and the subsequent increase in liver fibroblast density may play a role in the pathogenesis of alcoholic disease. PMID- 8408099 TI - Response of the rat mesenteric vasculature to chronic treatment with nitrendipine alone and in combination with atenolol: evidence of a significant drug interaction. AB - 1. Nitrendipine 3 mg kg-1 was administered alone and in combination with atenolol 50 mg mg-1 day for 21 days to male normotensive Wistar rats and to spontaneously hypertensive Japanese Okamoto rats. Blood pressure was monitored daily. 2. Systolic blood pressure was decreased in normotensive Wistars and SHRs by nitrendipine alone and in combination. In both cases the decrease was greater in the hypertensive animal. There was no evidence of the combination having an additive hypotensive effect. 3. Following treatment, the response to exogenous noradrenaline (NA) and periarterial nerve stimulation (PNS) was measured in the in situ blood perfused mesentery. 4. Treatment with nitrendipine and the combination reduced the response to exogenous noradrenaline; with both, the reduction was greater in the hypertensive animal. The combination produced a larger reduction in response than nitrendipine alone. 5. Treatment with nitrendipine alone reduced the response to PNS in both normotensive and hypertensive animals, although this effect was greater in the SHR. 6. Combination treatment failed to change the response to electrical stimulation in the SHR, while in the normotensive rat it resulted in a large increase in response to higher frequency (16 and 35 Hz) stimulation. 7. As nitrendipine given alone reduced the response to PNS, and as we have previously shown a similar effect with atenolol given alone (Draper, Kingsbury, Redfern & Todd, 1992), the effect of the combination of nitrendipine and atenolol on responses to PNS is apparently influenced by some interaction between the two drugs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408100 TI - Mapping of glutaraldehyde-treated bovine pericardium and tissue selection for bioprosthetic heart valves. AB - Glutaraldehyde-crosslinked bovine pericardium is widely used in bioprosthetic heart valve fabrication. In an attempt to set a scientific basis for more reproducible tissue selection, we produced and analyzed topographical maps of glutaraldehyde-treated bovine pericardium. Whole pericardia were divided into specific anatomical areas and their thickness was measured and mapped on templates. In each area, the suture holding power was determined in both parallel and perpendicular (to the base-apex line) directions; analyses of the tearing patterns in each fragment were used to evaluate predominant fiber orientation, and observations were confirmed by polarized light microscopy. Complete maps were superimposed graphically to aid in the selection of certain areas that would have known fiber orientation, high suture holding power, and suitable thickness. Our results describe regional heterogeneity of bovine pericardial structure and mechanical properties, specifically demonstrating variations in thickness, suture holding power, and collagen fiber orientation. Two areas of choice (representing about 35% of the total) were described as suitable for use in bioprosthetic heart valve fabrication. PMID- 8408101 TI - Mechanical and histological evaluation of amorphous calcium phosphate and poorly crystallized hydroxyapatite coatings on titanium implants. AB - The effect of amorphous calcium phosphate (Ca/P) and poorly crystallized (60% crystalline) hydroxyapatite (HA) coatings on bone fixation to "smooth" and "rough" (Ti-6A1-4V powder sprayed) titanium-6Al-4V (Ti) implants was investigated. Implants were evaluated histologically, mechanically, and by scanning electron microscopy (SEM) after 4 and 12 weeks of implantation in a rabbit transcortical femoral model. Histological evaluation of amorphous vs. poorly crystallized HA coatings showed significant differences in bone apposition (for rough-coated implants only) and coating resorption (for smooth- and rough coated implants) that were increased within cortical compared to cancellous bone. The poorly crystallized HA coatings showed most degradation and least bone apposition. Mechanical evaluation, however, showed no significant differences in push-out shear strengths between the two types of coatings evaluated. Differences between 4 and 12 weeks were significant for coating resorption and push-out shear strength but not for bone apposition. Significant enhancement in interfacial shear strengths for bioceramic coated as compared to uncoated implants were seen for smooth-surfaced implants (3.5-5 times greater) but not for rough-surfaced implants at 4 and 12 weeks. Rough implants showed greater mean interfacial strengths than uncoated smooth implants at 4 and 12 weeks (seven times greater) and to coated smooth implants at 12 weeks only (two times greater). Mechanical failure of the bone/coating/implant interface consistently occurred within the bone, even in the case of the poorly crystallized HA coatings, which had almost completely resorbed on rough implants. These results suggest that once early osteointegration is achieved biodegradation of a bioactive coating should not be detrimental to the bone/coating/implant fixation. PMID- 8408102 TI - Time-dependent changes in the mechanical properties of zirconia ceramic. AB - Zirconia has received special attention, mainly because of its high strength and toughness. However, there is some controversy about the time-dependent deterioration of its mechanical properties. To examine the change in mechanical properties of zirconia ceramics in vivo and in vitro, tetragonal zirconia polycrystal pieces were introduced into the medullary cavity of the tibia in Japanese rabbits and animals were sacrificed after 2, 4 and 6 weeks and 6, 12, and 30 mo, respectively. Alumina ceramic and hydroxyapatite (HAP) pieces were used as controls to investigate the differences in biocompatibility. Zirconia showed a bending strength of over 1000 MPa initially, and little time-dependent change in strength was found in both in vivo and in vitro environments. x-Ray analysis showed little change in the transformation rate, i.e., less than 5 mol % in vivo and in vitro over a period of 3 years. To estimate time-dependent changes in zirconia over a longer period, zirconia pieces were placed in 95 degrees C saline solution for over 3 years and their mechanical properties examined at chosen intervals. No serious decrease of bending strength was found over the 3 year period under these conditions. It is concluded that zirconia can be used clinically because it retains a bending strength of over 700 MPa under any experimental conditions for over 3 years. PMID- 8408103 TI - Effect of polyol type on the surface structure of sulfonate-containing polyurethanes. AB - Polyurethanes based upon polytetramethylene oxide (PTMO) as the polyol and derivatized with propyl sulfonate functionality pendant from the urethane nitrogen have previously been shown to possess good blood-contacting properties. Other investigators have shown that sulfonated polyurethanes containing polyethylene oxide (PEO) as the soft segment are much more thrombogenic than those containing PTMO as the soft segment. In this article, the surface properties of sulfonated polyurethanes based upon either PTMO or PEO are compared. Dynamic contact angle measurements show a significant decrease in the receding angles of the sulfonated PTMO-containing polyurethane as compared to its nonsulfonated precursor polymer. No significant difference is seen between the receding contact angles of either the sulfonated PEO-based polyurethane or its nonsulfonated analog. Variable-angle electron spectroscopy for chemical analysis (ESCA) studies of sulfonated PTMO-based polyurethane performed at room temperature show that there is a significant decrease in sulfur content at the surface. In contrast, the sulfonated PEO-based polyurethane showed little change in sulfur content with take-off angle. Finally, ESCA studies of freeze-dried surfaces show a significant increase in sulfur near the surface of the sulfonated PTMO-based polymer as compared to vacuum-dried samples but show no such increase for the sulfonated PEO-based polyurethane. It is suggested that the ability of the sulfonate functionality to be expressed at the surface may explain the observed differences in blood compatibility between the sulfonated polyurethanes based upon polyols of varying hydrophilicity. PMID- 8408104 TI - Hardness and Young's modulus determined by nanoindentation technique of filler particles of dental restorative materials compared with human enamel. AB - The recently developed nanoindentation technique was used to measure hardness and Young's modulus of small filler particles in resin composites and other dental restoratives. This technique eliminates the need to visualize indentations. Load and displacement are continuously monitored during a loading-unloading sequence, and hardness as well as Young's modulus are then calculated from the load displacement curves taking into account the geometry of the indenter. Thirteen posterior composites, 3 dental ceramics for CAD/CAM restorations, 1 sintered porcelain, and 1 amalgam were investigated in this study. The results were compared to the hardness and Young's modulus determined by nanoindentation of human enamel. Of the dental materials tested, only five materials contain inorganic filler particles with a nanohardness not statistically different from that of enamel. The predominant fillers in all other materials, except amalgam and the prepolymerized resin fillers in Bell Firm PX, were found to be significantly harder. The dental restorative materials, except the alloy phase in amalgam, were composed of particles with a Young's modulus significantly lower than that of human enamel. The alloy phase in amalgam had a Young's modulus value comparable to that of enamel. PMID- 8408105 TI - Collagen types I and III at the implant/tissue interface. AB - Collagen composition in tissue capsules around implants has been reported to differ histologically from collagen in subcutaneous connective tissue. In the present study, an immune histochemical analysis of collagen types I and III was undertaken in tissue capsules of various implant materials. The materials included polyvinyl chloride/polyacrylonitrile copolymer, poly(ethylene terephthalate), polysiloxane, titanium, and hydroxyapatite, which had been implanted into the dorsal subcutaneous space of rabbits for various time periods from 28 and to 90 days. The results indicate that collagen type III stained in all capsules independent of the evaluated materials, implantation periods, and material surface roughness. Collagen type I stained only in titanium implant capsules and dominated there over collagen type III. The staining sensitivity was highly specific and reproducible. The presence of collagen type III can be expected because it is the collagen of connective tissue healing. Collagen type I appears to be a response to chemical or electrochemical titanium surface properties but not to surface roughness. The quantitative relationship between the two collagen types may indicate capsule tissue stability and therefore serve as another biocompatibility measure. PMID- 8408106 TI - Osseous implant for studies of biomaterials using an in vivo electrochemical transducer. AB - The in vitro and in vivo electrochemical behavior of commercially pure titanium (cp Ti) was characterized using a specialized osseous implant in conjunction with electrochemical impedance spectroscopy (EIS) measurement techniques. Studies performed in vitro were used to verify the operation of the transducer and develop methods of deconvoluting EIS data. This method was subsequently used to describe an electrochemical equivalent circuit model of the surface oxide and electrical double-layer capacitance of cp Ti in the endogenous electrolyte found in the medullary compartment of a baboon tibia. Kinetic profiles of the double layer capacitance and the polarization resistance were constructed from multiple in vitro and in vivo EIS measurements performed over 60 min at 0 V (reference Ag/AgCI) conditioning potential. The profiles demonstrated that the growth of surface oxides was biphasic, with rapid decrease in the double-layer capacitance occurring within 20 min and reaching steady-state conditions at approximately 40 min. These data suggested that a passive, stable biofilm formed on the cp Ti surface in vivo and in vitro. PMID- 8408107 TI - In vitro study on the integrity of a hydroxylapatite coating when challenged with staphylococci. AB - An in vitro study was performed to evaluate the effect of Staphylococcus aureus and Staphylococcus epidermidis on the integrity of a hydroxylapatite coating. The coating plasma-sprayed on poly(L-lactide), showed dissolution during a 24 h incubation period. This was indicated by an increase in pH and calcium release in the buffer solution. After 4 h of incubation, calcium levels decreased due to the precipitation of calcium phosphate complexes on the coating. The bacteria digested or dissolved the coating, creating irregularly shaped holes. Although the integrity of the hydroxylapatite coating was focally damaged within 2-4 h of incubation with staphylococci, the extent of the damage was only marginal. Due to the formation of a layer of CaP precipitates though, bacteria could not be counted accurately after 4-8 h of incubation. This model could reveal part of the failure mechanism of infected hydroxylapatite coated implants. PMID- 8408108 TI - Particle microelectrophoresis of calcium-deficient hydroxyapatite: solution composition and kinetic effects. AB - Concurrent work demonstrates that the zeta potential of bone is multivalued and systematically alterable by changes in sample preparation, steeping fluid composition, and steeping time. Since bone mineral is a mixture of carbonated calcium-deficient hydroxyapatites, and since the zeta potential of calcium deficient hydroxyapatite (CDHA) is altered by pH and time in HNO3-KOH solutions, the zeta potential of CDHA in physiologic Neuman's fluid (NF) compared with that seen in bone could reveal important information on the contribution of the mineral phase to the zeta potential of bone. In addition, such information may be valuable in designing and evaluating calcium-phosphate ceramics for increased bone ingrowth. Results demonstrate that the zeta potential of CDHA in NF is negative. With increasing calcium in NF, the zeta potential magnitude of CDHA decreases and inverts to positive values given sufficient calcium concentration and steeping time. This result is opposite to that seen in bone, suggesting that exposed CDHA is not the predominant bone microsurface and implicating a bone surface protein component. With increasing phosphate in NF, the zeta potential magnitude increases to more negative values. While low concentrations of fluoride showed no effect, the possibility of an effect with higher concentrations is still to be determined. PMID- 8408109 TI - Osteogenesis at the dental implant interface: high-voltage electron microscopic and conventional transmission electron microscopic observations. AB - The osteogenesis of mandibular bone to endosteal dental implants was examined using an in vivo dog model. One half of the implants examined were unloaded implants, with the remaining one half prosthodontically loaded for 6 months. Undecalcified mandibular implant samples were examined with both high-voltage electron microscopy (HVEM) stereology and routine transmission electron microscopy. The osseous interface to integrated implants was shown to vary in its morphology. Mineralized bone was observed directly apposing the implant, often separated from the implant by an electron-dense deposit of approximately 50 nm. Within this densely mineralized matrix, osteocytes were routinely observed. Adjacent areas were shown to contain slightly wider zones of either a less dense mineralized matrix or, alternatively, unmineralized tissue. Other zones consisted of wider unmineralized matrices containing collagen fibers and osteoblasts. These latter zones were consistent with the appearance of an appositional type of bone growth. Because bone is a dynamic, actively remodeling tissue, a varied morphology of the support tissues to dental implant is not unexpected. Areas of mature bone interfacing with successfully integrated implants were demonstrated, as well as areas adjacent to the mature bone that were undergoing remodeling or mineralization. This study has also shown that HVEM stereology is a valuable research tool to investigate the oral tissue interface with dental implants. PMID- 8408110 TI - Elastomeric polytetrapeptide matrices: hydrophobicity dependence of cell attachment from adhesive (GGIP)n to nonadhesive (GGAP)n even in serum. AB - The cross-linked polytetrapeptide matrices based on the repeating amino acid sequences, GGAP, GGVP and GGIP, were prepared and tested for cell adhesion promoting activity in both the absence and presence of fetal bovine serum. For comparison, X20-poly(GVGVP), a matrix previously shown to be a poor support for cell attachment and spreading, was included. In the absence of serum, all three polytetrapeptide-based matrices and the polypentapeptide-based matrix were negative for the adhesion of fibroblasts and endothelial cells. In the presence of serum, various sub-maximal levels of cell adhesion were found for all matrices except for the matrix based on GGAP. An apparent correlation was noted between the degree of cell attachment to the different polytetrapeptide-based matrices and the hydrophobicity of those matrices where increased hydrophobicity results in increased cell attachment. The property of being refractory to ligamentum nuchae fibroblast and human umbilical vein endothelial cell adhesion in the presence of serum indicates a potential use for X20-poly(GGAP) in the development of, for example, additional physical barriers for the prevention of post-surgical and post-trauma adhesions. PMID- 8408111 TI - Immobilization of high-affinity heparin oligosaccharides to radiofrequency plasma modified polyethylene. AB - Oligosaccharides of heparin with high affinity for antithrombin III (ATIII) have been immobilized onto surface-modified NHLBI Primary Reference low density polyethylene (PE). PE was modified by radiofrequency plasma polymerized (< 150 nm thick) films derived from N-vinyl-2-pyrrolidone (PPNVP) or allyl alcohol (PPAA), and coupled by chemical derivatization to either 3-aminopropyltriethoxysilane or amino-terminated poly(ethylene oxide). High affinity heparin oligosaccharides (HA heparin, anti-factor Xa activity of 592 +/- 120 IU/mg) prepared by partial deaminative cleavage of commercial crude heparin and fractionated by agarose ATIII affinity chromatography, were immobilized to surface-modified PE by reductive amination. The anticoagulant activity, as determined by a chromogenic assay for the inhibition of factor Xa, was estimated to be 30-70 mIU/cm2, with binding estimated to be 56-119 ng/cm2. The highest activity was obtained for the HA-heparin immobilized to PE modified by PPNVP with a PEO spacer. Visual confirmation of ATIII binding to immobilized HA-heparin was demonstrated by a gold-labeled double antibody method with imaging by SEM. PMID- 8408112 TI - Detection of remnant proteolytic activities in unimplanted glutaraldehyde-treated bovine pericardium and explanted cardiac bioprostheses. AB - The presence and activity of proteolytic enzymes has been investigated in vitro on soluble and insoluble preparations obtained from both unimplanted and implanted glutaraldehyde-treated bovine parietal pericardium. Using detection by colorimetric techniques, soluble preparations were shown to hydrolyze enzyme substrates that are characteristic for trypsin-like proteases, cathepsin-like proteases, and collagenase. As detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in gradient gels and gel filtration on Sepharose CL-6B, insoluble (pellet) preparations degraded denatured type I collagen in a time dependent pattern, producing low-molecular-weight fragments. These activities were partially inhibited by phenylmethylsulfonyl fluoride, N-ethyl maleimide, soybean trypsin inhibitor, para-chloromercuribenzoic acid, or ethylenediaminetetraacetic acid, suggesting the presence of a heterogeneous enzymatic mixture. Insoluble preparations incubated with pure pericardial dermatan sulfate proteoglycan detached the glycosaminoglycan chains from their core protein carrier, producing a digestion pattern similar to Cathepsin C. These findings demonstrate the presence of active proteases in glutaraldehyde-fixed bovine pericardium per se and in explanted pericardial bioprosthetic cardiac valves, an additional factor that might contribute to intrinsic extracellular matrix degeneration in pericardial bioprosthetic devices. PMID- 8408113 TI - In vivo transport and excretion of corrosion products from accelerated anodic corrosion of porous coated F75 alloy. AB - Corrosion of porous coated cobalt chromium specimens surgically implanted subcutaneously in hamsters was accelerated by application of a fixed anodic potential. Corrosion current measurements were utilized for prediction of metal ion release based on Faraday's law. Daily urine samples were collected pre- and post-stimulation, and, at animal sacrifice, organs and blood samples were taken for chemical analysis. By using in vitro release data from previous studies, and efficiency correction factors, the amount of metal ion release was calculated. The results demonstrated rapid and complete excretion of nickel and molybdenum. Most of the cobalt was excreted, with elevation of cobalt levels in liver, kidney, and lung. Chromium excretion was much lower due to significant red cell binding and in vivo storage, especially in the kidney and spleen. PMID- 8408114 TI - Adsorption/desorption of human serum albumin at the surface of poly(lactic acid) nanoparticles prepared by a solvent evaporation process. AB - Biocompatible and biodegradable nanoparticles of poly(lactic acid) (100% L-lactic units = PLA) were prepared by an emulsion, microfluidization, and solvent evaporation method using human serum albumin (HSA) as a surface agent. A radiolabeling technique was employed to quantify the serum albumin bound to the nanoparticles and to measure its desorption kinetics in various media at 22 degrees C and 37 degrees C (phosphate buffer pH 7.4, serum albumin 40 g/L in phosphate buffer pH 7.4 and fetal calf serum). The amount of serum albumin bound to the nanoparticles was found to be a linear function of 1/D (where D is the nanoparticle mean diameter) and was related to the total developed area of the nanoparticles. The adsorption/desorption behavior of serum albumin at the surface of the nanoparticles suggested a multilayer adsorption model. Moreover, a part of the serum albumin molecules was irreversibly bound regardless of the incubation conditions. Consequently, the classical Langmuirian theories of equilibria could not be applied. PMID- 8408115 TI - Effects of hydroxyapatite implants on primary mineralization during rat tibial healing: biochemical and morphometric analyses. AB - The effect of 40- to 60-mesh hydroxyapatite (HA) granules (Calcitek, Inc., Carlsbad, CA) on the process of primary mineralization during bone healing was examined following insertion of the HA granules into rat tibial bone after marrow ablation. Response to HA was assessed by monitoring morphometric and biochemical changes in matrix vesicles, which are extracellular organelles associated with initial calcification. Following insertion of HA, matrix vesicle-enriched membranes (MVEMs) were isolated from the tissue adjacent to the implant and from the endosteum of the contralateral limb at 3, 6, 14, and 21 days and from a nonimplanted control group (t = 0). MVEM alkaline phosphatase- and phospholipase A2-specific activities were increased on days 6 (peak) and 14; phosphatidylserine content was also elevated on days 6 and 14 (peak). Comparable changes were seen in the contralateral limb but at lesser magnitudes. Morphological changes were observed as well. The number of matrix vesicles/micron2 matrix increased on days 6 (peak) and 14. The mean diameter of the matrix vesicles was elevated on days 6 (peak), 14, and 21. Mean distance from the calcifying front increased on day 6 but was decreased on days 14 and 21. These results indicated that HA behaves like bone-bonding implants in that there is a stimulation of matrix vesicle enzymes, increased phosphatidylserine content, and increase numbers of matrix vesicles. However, the increases occur only after 6 days postimplantation, indicating a delay in response when compared to normal healing. This delay is confirmed by the morphometric measurements. HA causes a reduction in the response associated with marrow ablation. In addition, the effects of HA are comparable locally and systemically but with different intensity. These observations suggest that osteogenic cells are able to compensate for the inhibitory effects of HA and primary calcification involves normal matrix vesicle production and maturation, if somewhat delayed and reduced in magnitude. The ability to support primary mineral formation may contribute to the successful bonding of HA with surrounding osseous tissue. PMID- 8408116 TI - The effects of bone cement powder on human adherent monocytes/macrophages in vitro. AB - This study reports the effects of Simplex bone cement powder (BC) on the proliferation and production of bone resorbing factors in vitro by human adherent monocytes/macrophages. Adherent peripheral blood cells were isolated from seven healthy individuals and exposed to a dispersion of BC powder (1 mg/mL), phytohemagglutinin (PHA, 40 micrograms/mL), or medium alone at different periods of cell incubation (days 0-2, 0-7, 5-7, or 10-12). Cell proliferation was quantified by incorporation of 3H-thymidine uptake. Culture supernatants were evaluated for levels of interleukin 1-like activity (IL-1) by murine thymocyte proliferation assay, prostaglandin E2 (PGE2) by radioimmunoassay, lysosomal enzyme activity (N-acetyl-beta-D-glucosaminidase and beta-glucuronidase using fluorometry, and collagen and casein degrading activity using radioactive substrates. Human adherent peripheral blood cells showed a proliferative response to PHA that coincided with cell maturation; BC did not inhibit PHA-induced cell proliferation of either adherent or nonadherent blood cells, indicating the non toxic nature of these particles at the concentrations tested. BC stimulated increased release of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase; the levels of PGE2, IL-1, collagenase, and caseinase were unchanged. PMID- 8408117 TI - Mechanisms and structure of the bond between bone and hydroxyapatite ceramics. AB - Two assays were carried out to investigate the postulates that dissolution/reprecipitation phenomena and epistaxis are involved in the formation of the bond between sintered hydroxyapatite (HA) and bone. HA was exposed to a physiologic solution and the ions going into solution quantified using inductively coupled plasma (ICP) emission spectroscopy. The HA was retrieved and examined using scanning electron microscopy (SEM). In the second assay, HA was implanted into Alsatian femura. Following retrieval, specimens were prepared and studied with SEM, at magnifications usually reserved for transmission electron microscopy (TEM). In vitro the HA surface composition appeared to shift to the acidic (calcium-deficient) direction. The HA surfaces in both assays underwent a dissolution/reprecipitation degradation of varying severity, forming a recrystallization layer of spherocrystallites. Two modes of bone bonding to implanted HA were identified: (1) bone tissue components bonded to HA via a recrystallization zone similar in structure to the reprecipitation layer in the corrosion assay, and (2) bone tissue components bonded directly to HA crystals with no morphologically discernible signs of dissolution embarrassment. The formation of the bone/HA bond seems to involve dissolution/reprecipitation phenomena. What is believed to be the first morphological evidence of epitaxial growth involvement in the formation of this bond is presented. PMID- 8408118 TI - Quantification of in vitro endothelial cell adhesion to vascular graft material. AB - This study tests the hypothesis that denucleating vascular graft material and binding cell adhesion molecules increases endothelial attachment. Removal of gas nuclei (denucleation) increases the available surface area of biomaterials for modification and/or cell adhesion, while adhesion molecules provide specific attachment sites. Microvascular endothelial cells (MVEC) were isolated from fat, fluorescently labeled, and allowed to settle onto expanded polytetrafluoroethylene (ePTFE) vascular patches. Patch treatments included fibronectin alone (F), gas denucleation followed by fibronectin (D/F), denucleation followed by the surfactant tridodecylmethylammonium chloride (TDMAC) (D/T), denucleation followed by TDMAC followed by fibronectin (D/T/F), or denucleation followed by TDMAC followed by a synthetic polymer with numerous arginine-glycine-aspartic acid sequences (D/T/R). After 1 h of incubation, the 45 mm2 patch area covered with microvascular endothelial cells was assessed using computer-aided fluorescence microscopy. Initial graft coverage with D/T (26.2 +/- 2.4 mm2) and D/T/F (25.9 +/- 2.1 mm2) was better than with F (16.8 +/- 2.5 mm2) (P < .05). Patches were then exposed to a detachment stress and coverage was again measured. Following stress, coverage was greater with D/T (20.7 +/- 3.4 mm2) and D/T/F (20.7 +/- 2.0 mm2) than with D/T/R (8.4 +/- 1.8 mm2) or F (3.6 +/- 0.9 mm2) (P < .001). Percent retention of cells following stress was better with D/T and D/T/F than with D/T/R, D/F, or F (P < .0001). Scanning electron micrographs were consistent with the qualitative findings. The results indicate that TDMAC alone or with fibronectin increases adhesion of human microvascular endothelial cells to denucleated ePTFE. PMID- 8408119 TI - In vivo biostability and calcification-resistance of surface-modified PU-PEO-SO3. AB - To examine the biostability and calcification-resistance of polyurethanes (PUs), the surface of PU was grafted with hydrophobic perfluorodecanoic acid (PFDA) (PU PFDA), hydrophilic polyethyleneoxide (PEO) (PU-PEO1000), and further negatively charged sulfonate groups (PU-PEO1000-SO3). An in vivo animal test was conducted by subcutaneous implantation in rats during 2, 4 and 6 months. A scanning electron microscope study demonstrated that the degree of surface cracking on explanted PUs was increased in the following order: PU-PFDA > PU > PU-PEO1000 > PU-PEO1000-SO3. In the results of energy dispersive x-ray analysis and inductively coupled plasma atomic emission spectrometry, the deposition of calcium was found abundantly, but that of phosphorus was hardly in existence in all implanted PUs, suggesting that this calcium compound is not a hydroxyapatite. The calcium contents, regardless of implantation time, were also increased in the same order (PU-PFDA > PU > PU-PEO1000 > PU-PEO1000-SO3). After 6 months implantation, no severe tissue reactions were observed and calcification almost occurred on polymer surfaces in all implants. Such superior biostability and anticalcification of PU-PEO1000-SO3 might be attributed to synergistic effects of its excellent surface smoothness, sulfonate acid (SO3-) groups, nonadhesive and mobile PEO, and the high hydrophilicity and enhanced blood compatibility. Therefore, PU-PEO1000-SO3 is promising as biostable and calcification-resistant biomaterial. PMID- 8408120 TI - Magnitude and polarity of a fluoroethylene propylene electret substrate charge influences neurite outgrowth in vitro. AB - Positively charged coating materials such as polylysine improve neuronal attachment in vitro. Due to the structural complexity of these charged molecules, it is unclear whether neuronal effects are due to charge or to physicochemical effects, or both. Polymeric materials with charge storage capabilities and defined surface properties may provide a model in which electrical charge and surface property effects can be separated. Fluorinated ethylenepropylene (FEP) films can store negative or positive charges injected through a corona charging process, thus generating a negative or positive external electrostatic field. In the present study, mouse neuroblastoma (Nb2a) cells were cultured on positive, negative, and uncharged FEP substrates, in both serum-containing and serum-free media. Cell attachment, differentiation, and neurite outgrowth were assessed 24, 48, 72 and 96 h after plating. Electron spectroscopy for chemical analysis (ESCA), contact angle analysis, and scanning electron microscopy (SEM) revealed no differences in surface chemistry and topography between positive, negative, and uncharged FEP. No significant differences in the levels of cell attachment on positive, negative, and uncharged substrates were observed. Significantly higher levels of neurite outgrowth, however, were observed with positive substrates as compared to negative and uncharged substrates, in both media conditions. Substrates charged to +1000 V showed greater levels of outgrowth compared to +500 and +3000 V, suggesting the presence of an optimal range of charge for neurite outgrowth. These results show that cell/charge interactions mediate cell effects on electrically charged substrates with identical surface chemistry, topography and adhesivity. PMID- 8408121 TI - Dacron vascular biomaterial triggers macrophage ectoenzyme activity without change in cell membrane fluidity. AB - Biomaterials induce an inflammatory reaction characterized by a rapid recruitment at the implantation site of polymorphonuclear cells and macrophages. In the course of the inflammatory response, the cellular activation triggers expression of a number of enzymes, such as 5'-nucleotidase, which is widely distributed in animal cell membranes as an ectoenzyme. It is now well established that 5' nucleotidase activity decreases following the contact of inflammatory cells with foreign particles. In this paper we investigate a possible correlation between the enzymatic activities and the dynamic properties of the cell membrane bilayer. Dacron pieces were introduced into rats' peritoneal cavities for a period of 6 h, after which the peritoneal cells were harvested, and various enzyme assays performed, including those for cytoplasmic, lysosomal, and ectoenzymes. In parallel, we studied cell membrane fluidity, using fluorescence polarization of 1 (4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH), and cellular ultrastructural alteration resulting from the cell-biomaterial interactions using scanning and transmission electron microscopy. Our results show that: 1) macrophages spread around the Dacron fibers with cytoplasmic finger-like projections, but no phagolysosomes, 2) 5'-nucleotidase levels decrease with surgical trauma in comparison with the resident cell exudate, 3) implantation of biomaterials slightly modify the 5'-nucleotidase levels observed in the sham animal, 4) no differences in the anisotropy values indicating that membrane lipid order within the cells could not account for the observed decrease of 5' nucleotidase activity. Thus, we can suggest that 5'-nucleotidase expression may reflect a particular feature of cell activation without a phagocytic process. PMID- 8408122 TI - Effects of Na2HPO4 and NaH2PO4 on hydroxyapatite formation. AB - Particulate solids CaHPO4 (DCP) and Ca4(PO4)2O (TetCP) can react to form hydroxyapatite (HAp) under conditions which may be compatible with those in vivo. Effects of sodium phosphate solutions on the formation of calcium deficient HAp at 25 degrees C by this reaction in dilute suspensions (liquid-to-solids ratio of 100) and at 1/s = 1 were studied. An 1/s ratio of one is not greatly different from that at which monolithic HAp forms. Solution chemistry results showed that the sodium phosphate used and its concentration strongly influenced the kinetics of HAp formation and the HAp composition. HAp formation occurred by the same mechanism in water and in the Na2HPO4 solutions investigated; however, HAp formation occurred more rapidly in Na2HPO4 solutions. DCP dissolution was congruent in these solutions and this delayed the overgrowth of DCP by HAp. Reaction in NaH2PO4 solutions resulted in the formation of CaHPO4.2H2O as an intermediate product. The final product formed in the NaH2PO4 solutions was affected by the l/s ratio used. CaHPO4.2H2O, not HAp, was the product formed from dilute suspension but it was not observed during reaction at l/s = 1. Variations in solution were correlated with the formation of HAp and intermediate products, with the consumption of reactants, and with the heat evolution characteristics. The latter were determined by isothermal calorimetry. Calorimetric analysis of the reactions indicated increased heat evolution in sodium phosphate solutions and acceleration in HAp formation. This appears to be related to accelerated dissolution of CaHPO4, which was rate limiting. PMID- 8408123 TI - Effect of the conformation and orientation of adsorbed fibronectin on endothelial cell spreading and the strength of adhesion. AB - The effect of surface hydrophobicity upon the conformation of the cell binding domain of fibronectin (Fn) and the influence of Fn conformation on bovine aortic endothelial cell (BAEC) adhesion were examined. The free sulfhydryl group of Fn located near the cell binding domain was selectively labeled with acrylodan, a polarity sensitive fluor. Fluorescence emission was monitored in solution and upon adsorption to hydrophilic glass and hydrophobic silanized glass. The acrylodan-labeled Fn emission maximum shifted to longer wavelengths upon adsorption and the shift was greater for acrylodan-labeled Fn adsorbed to hydrophilic glass than hydrophobic silane, suggesting that the acrylodan was in a more solvent accessible environment on glass than silane. BAEC, suspended in serum-free medium, attached for 15 or 120 min onto glass or silane surfaces containing preadsorbed Fn, after which cell spreading and the strength of adhesion in a parallel plate flow chamber were measured. Cell spreading was similar on both surfaces after 15 min attachment, but BAECs were more spread on glass than silane after 120 min. At low surface concentrations of Fn, BAECs were more adherent on glass than silane. At higher surfaces concentrations, adhesion was similar. After a 2-h incubation in serum-free medium, cells on glass showed more extensive development of focal contacts as determined by immunofluorescent staining for vinculin. Cell adhesion under flow was reduced on silane by inhibition of protein synthesis with cycloheximide, suggesting that cell attachment to silane was promoted by cellular synthesis of Fn. The results indicate that changes in the conformation of the Fn cell binding domain affect Fn affinity for its cell surface receptor. PMID- 8408124 TI - Examination of efferent lymph nodes after 2 years of transcortical implantation of poly(L-lactide) containing plugs: a case report. PMID- 8408125 TI - Effect of the tooth microstructure on the shear bond strength of a dental composite. AB - Plane human enamel and dentin surfaces were used for microstructural investigation and shear bond testing. The dental tissue microstructure was characterized through the surface topology, the concentration in mineral elements (determined by electron microprobe analysis), and the Vickers microhardness for the same dental tissue. The etched prismatic enamel presents a surface roughness of about 200% of the apparent area, while the intertubular surface fraction of the dentin is, on the average, equal to 75%. The calcium concentration lies between 23.2 and 37.8% of the enamel total mineral content, and 18.5 and 28.2% of the dentin. The microhardness varies from 205 to 378 Hv for the enamel and from 37 to 98 Hv for the dentin. Also, the shear bond strength to a chemosetting adhesive/composite system varies from 10.4 to 23.9 MPa for the enamel and from 0.0 to 5.0 MPa for the dentin. It is shown that the shear bond strength is strongly correlated to the microhardness through a simple linear equation valid for both enamel and dentin surfaces. In turn, the microhardness is correlated to the calcium concentration through two distinct linear relations. These correlations lead to the intrinsic dependence of the shear bond strength on the calcium concentration of the dental tissue, providing that corrections are applied to the bond strength data in order to take into consideration the effective solid area of the adherend and the polymerization retraction stress. Consequently, it is proposed that the adhesion mechanisms for the both enamel and the dentin are controlled, to a major extent, by the mineral content and the surface topology of the tooth. PMID- 8408126 TI - Polyvinyl alcohol plasma deposited nylon 4 membrane for hemodialysis. AB - Polyvinyl alcohol (PVA) is a hydrophilic and blood compatible material, but it is easily hydrolyzed in aqueous solution. Various methods, including chemical crosslinking and gamma-ray irradiation, have been investigated by many researchers for overcoming the instability of PVA in aqueous solution. This study suggests a new hemodialysis membrane, which is prepared by plasma depositing vinylacetate (VAc) onto annealled nylon 4 membrane, followed by hydrolysis treatment. This improves the blood compatibility of nylon 4 and overcomes the hydrolysis problem of PVA. In addition to conventional plasma deposition, the plasma initiated interpenetrating network (IPN) method is also investigated. The new hemodialysis membranes prepared by both conventional plasma deposited VAc onto nylon 4 (PPVA/N4) and IPN polymer of plasma initiated VAc onto nylon 4 (PIPVA/N4 IPN) show significant improvement in blood compatibility. However, the solute permeabilities of PIPVA/N4 IPN membranes are higher than those of PPVA/N4 membranes. The permeabilities of NaCl, vitamin B12, and albumin for membranes prepared by IPN at 80 W, 30 sec plasma deposited conditions, i.e., the optimized conditions, are 3.614, 0.070, and 0.000 x 10(-5) cm2/min, respectively. The mole ratio of adsorbed fibrinogen to adsorbed albumin (the mole ratio F/A) for this membrane is 0.20, and it also shows excellent blood compatibility in this study. PMID- 8408127 TI - Determination of the 3-D morphology of degradable biopolymer implants undergoing in vivo resorption. AB - A technique is presented that allows the visualization of the degradation process of biopolymers. Serial thick-section specimens containing cross-sections of polydioxanone implants (PDAs) were realigned for 3-D reconstruction using a double-embedding technique. The outlines of the cross-sectioned implants were traced using an automatic image analysis system and converted into x, y, and z coordinates. The reconstruction of the implant body was achieved by the stepwise analysis of vertical relationships between two adjacent section levels. The surface was approximated by triangulation and calculated from the surface triangles. Visualization was achieved by introducing the data into commercially available software using z-buffering and Gouraud shading. The geometric precision of the reembedding technique was found to be 0.8-1.5% of the screen width; recovering experiments showed a good approximation to the actual physical volume of unresorbed implants (+/- 2.6%). The error of volume determination due to the limited resolution in the z direction was calculated to be acceptable (< 5.0%) in isotropic objects where the ratio between the radius of the surface curvature and the distance between the sections is > 4. Results indicate that large devices of PDAs are degraded in vivo in a fashion similar to that previously described for in vitro degradation of PLA molded screws by a surface/center differentiation with formation of hollow residuals after 17 weeks and complete degradation with phagocytosis of microparticles after 26 weeks. PMID- 8408128 TI - Apatite formation on three kinds of bioactive material at an early stage in vivo: a comparative study by transmission electron microscopy. AB - Apatite formation on the surface of three kinds of bioactive material at an early stage after implantation in bone was studied using transmission electron microscopy (TEM). The materials were apatite- and wollastonite-containing glass ceramic (A-W GC) as a surface-active glass-ceramic, dense sintered hydroxyapatite (HA) as a surface-active ceramic, and dense sintered beta-tricalcium phosphate (beta-TCP) as a resorbable ceramic. Particles of these materials, ranging from 100-300 microns in diameter, were implanted into rat tibiae, and specimens were prepared at 3, 7, 10, and 14 days after implantation. For A-W GC, dissolution of the glassy and probably wollastonite phase was observed in the surface region on and after the third day, and a collagen-free thin apatite layer on the surface of the material was evident on and after the seventh day. This apatite layer was observed before the mineralization of the surrounding bone matrix and was sometimes evident even where the material bordered on the bone marrow. On and after the tenth day, the surrounding bone matrix calcified and A-W GC-bone bonding through an apatite layer was completed. For HA, a mineralized collagen free layer was observed on the surface of the ceramic on and after the tenth day. This layer was always present near calcifying bone and it was difficult to distinguish from immature bone. For beta-TCP, such a surface mineralized layer was rarely evident, even just before bone-ceramic contact, and finally the bone bonded to beta-TCP directly. Cell-mediated degradation of beta-TCP was frequently observed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408129 TI - Alfred R. Shands, Jr., Lecture. The new Esperanto. PMID- 8408130 TI - The team player. PMID- 8408131 TI - Open tibial fractures with severe soft-tissue loss. Limb salvage compared with below-the-knee amputation. AB - The long-term outcomes and the quality of life were studied in patients who had had an open fracture of the tibial shaft with severe soft-tissue loss. Limb salvage with a free flap was attempted in twenty-seven patients (sixteen of whom had a successful procedure and were examined personally by us), while eighteen patients were managed concurrently with an early below-the-knee amputation. Soft tissue coverage was successful in all but one patient in whom limb salvage had been attempted. Ultimately, however, five extremities were amputated, with an infection at the site of a non-union being the most common reason for amputation. The patients who had had limb salvage had more complications (p < 0.001), more operative procedures (p < 0.001), and a longer stay in the hospital (p < 0.05) than the patients who had had an early below-the-knee amputation. The long-term functional results for sixteen patients who had had a successful limb-salvage procedure (average duration of follow-up, thirty-five months) were compared with those for eighteen patients who had had a below-the-knee amputation (average duration of follow-up, forty-four months). The patients who had had a successful limb-salvage procedure took significantly more time to achieve full weight bearing (p < 0.05), were less willing or able to work (p < 0.01), and had higher hospital charges (p < 0.006) than the patients who had been managed with an early below-the-knee amputation. They also had a significant decrease in motion at the ankle and subtalar joint in the injured leg compared with the contralateral leg (p < 0.001). A quality-of-life evaluation was possible for only thirteen of the patients who had had a successful limb-salvage procedure and for sixteen of the patients who had had a below-the-knee amputation. The two groups were similar in terms of their responses, but significantly more patients who had had limb salvage considered themselves severely disabled (p < 0.05). They also had more problems with the performance of occupational and recreational activities (p < 0.05). This study confirmed the reliability of modern microvascular free tissue techniques for the coverage of large soft-tissue defects associated with tibial fractures. It also showed that complications and difficulties in the restoration of osseous union are common and may be directly related to the less satisfactory functional, occupational, recreational, and quality-of-life outcomes that are seen in many patients who have had limb salvage.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408132 TI - Secondary reconstruction after vascularized fibular transfer. AB - We evaluated the results of skeletal reconstruction performed through a mature, vascularized fibular graft in five patients. The average time-interval between the original transplant and the secondary reconstruction was sixty-eight months. The indication for the initial graft had been the loss of bone secondary to trauma in one patient, a skeletal defect due to ablation of a tumor in two patients, and osseous loss due to resection of a congenital pseudarthrosis in two patients. The indication for the second reconstruction was non-union of a fracture as a result of a new traumatic injury in two patients and complex angular deformity in three patients; one of the patients in the latter group had an associated leg-length discrepancy. In all five patients, the second reconstruction was successful, and the vascularized fibular graft responded to the procedure in a manner similar to normal cortical bone. PMID- 8408133 TI - Functional analysis of patients who have had a modified Van Nes rotationplasty. AB - Electromyographic analysis of gait in eight patients who had had a modified Van Nes rotationplasty was performed to compare the activity of the flexors and extensors of the ankle on the side of the operation with that of the flexors and extensors of the knee on the normal, contralateral side. The resemblance in the activity of the extensors was more pronounced than that of the flexors. In three patients, the angle of flexion of the knee throughout the gait cycle was the same on the side of the rotationplasty as on the normal side. There was more symmetry in the swing phase than has been reported for patients who have had an above-the knee amputation. The strength of the dorsiflexors of the ankle on the side of the rotationplasty was 68 percent and that of the flexors was 71 per cent when compared with that of the muscles of the normal ankle. PMID- 8408134 TI - Ewing sarcoma of the pelvis. Clinicopathological features and treatment. AB - The results of treatment in twenty-seven patients who had a Ewing sarcoma of the pelvis were reviewed. Six patients had had metastatic disease at the time of the diagnosis. The three-year actuarial survival of these patients was 17 per cent (95 per cent confidence interval, 8 to 52 per cent). Of the twenty-one remaining patients, thirteen had received chemotherapy and radiation therapy to the primary lesion and eight had had chemotherapy and operative resection, with or without radiation therapy. The actuarial five-year over-all survival was 25 per cent (95 per cent confidence interval, 6 to 51 per cent) in the group that had had radiation without a resection and 75 per cent (95 per cent confidence interval, 31 to 93 per cent) in the group that had had a resection (p < 0.005, log-rank method). The actuarial over-all five-year survival was 45 per cent (23 to 65 per cent) for all patients who had had localized disease when first seen. Actuarial local failure analysis (the censoring of patients who died without evidence of local failure before the two-year follow-up examination) revealed a rate of local failure of 44 per cent (14 to 79 per cent) in the group that had been treated with chemotherapy and radiation alone compared with 13 per cent (0 to 53 per cent) in the patients who had had a resection, but this difference was not significant (p > 0.25, log-rank method).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408135 TI - Treatment of giant-cell tumor of the pelvis. AB - Nineteen patients who had a giant-cell tumor of the pelvis were managed from 1944 through 1989. Thirteen of the patients were in the third or fourth decade of life. The ileum was involved in thirteen patients; the pubis, in two; the ischium, in three; and the pubis and ischium, in one. Five patients who had an initial Stage-II (active) lesion were managed with curettage; in one of the five, a local recurrence developed at twelve months, and the other four had no evidence of disease from nine to forty-six years after the curettage. Eight patients who had a Stage-III (aggressive) lesion were managed with resection of most of the lesion followed by curettage of any remaining positive margins; four of the eight also received adjuvant irradiation. None of the eight had a local recurrence; six had no evidence of disease from eight to twenty-seven years after the operation, one died because of a metastatic post-radiation sarcoma thirteen years after the operation, and one was asymptomatic but had pulmonary metastases three years after the operation. Four patients who had a Stage-III recurrent local tumor when they were first seen were managed with hemipelvectomy, wide resection, resection and curettage, and curettage and arthroplasty (one procedure each). Three had no evidence of disease seven to twenty years after the operation and one had died because of a post-radiation metastatic osteosarcoma eight years after the operation. Two patients who had Stage-III disease were managed with external beam radiation alone. One had no evidence of disease twenty-six years later. The other had a recurrence one year later, which was treated with additional irradiation; a second recurrence was treated with curettage and bone-grafting. Twenty-eight years after the initial radiation treatment, this patient had no evidence of disease. External beam irradiation was used for a total of eight patients; a post radiation sarcoma developed in two. PMID- 8408136 TI - Operative treatment of sacrococcygeal chordoma. A review of twenty-one cases. AB - Between 1972 and 1992, twenty-one patients had a primary operation for the treatment of a sacrococcygeal chordoma; seventeen had had a diagnostic biopsy elsewhere. The average age at the time of the operation was fifty-five years (range, six to seventy-eight years); fourteen patients were male and seven were female. In all patients, a posterior approach was used, even for resections at the cephalic levels of the sacrum. In addition, sixteen of the twenty-one patients were treated with adjuvant radiation therapy. Four patients died; three died of metastatic chordoma. Of the remaining seventeen patients, fifteen were apparently free of disease and had not had a local recurrence at the time of the latest follow-up examination. The average duration of follow-up for these fifteen patients was four and one-half years. Of the nine patients who were followed for at least five years, seven were disease-free at the latest follow-up evaluation. Of the seven patients in whom both second sacral roots were the most caudad nerve roots spared, four had normal bladder control and five had normal bowel control. Of the four patients in whom the most caudad nerve-roots spared were the first sacral or more cephalic roots, all had impaired bladder control, one had impaired bowel control, and three had a colostomy. PMID- 8408137 TI - Symptomatic synovial plicae of the knee. AB - Forty-five knees (thirty patients) with a specific diagnosis of synovial plica syndrome, and without any other known lesion, were randomized to be treated with either diagnostic arthroscopy alone or arthroscopy and division of all plicae. The diagnosis of synovial plica syndrome had been made on the basis of intermittent pain in the anterior aspect of the knee, painful clicking with activity, giving-way, and a palpable, tender plica. The patients were selected for arthroscopy only if the symptoms had continued unabated after a course of physical therapy. At the time of follow-up, improvement had occurred in only six (29 per cent) of the twenty-one knees in which the plicae had not been divided, in contrast with twenty (83 per cent) of the twenty-four knees in which they had been divided (p < 0.001). Ten (48 per cent) of the knees in which arthroscopic division had not been done were treated with another arthroscopic operation. Seven of these ten knees improved after the subsequent division of the plicae (p < 0.01). We concluded that synovial plicae of the knee may be a definite cause of anterior pain in children and adolescents. PMID- 8408138 TI - Custom-designed femoral prostheses in total hip arthroplasty done with cement for severe dysplasia of the hip. AB - A custom-designed femoral prosthesis was implanted with cement and a standard acetabular component was used to treat nineteen severely dysplastic hips in fourteen consecutively managed patients. Components that had been custom-designed with the use of plain radiography were used because the anatomical reconstructive goals could not be achieved with commercially available implants. These goals were to match the offset of the femoral head and the length of the lower limb with those on the normal side for patients who had unilateral involvement and to provide an average (thirty to forty-millimeter) offset with equal limb lengths for patients who had bilateral involvement. A retrospective clinical and radiographic analysis was performed. The diagnoses included coxa vara (one hip), congenital dislocation (twelve hips), achondroplasia (three hips), and spondyloepiphyseal dysplasia (three hips). The mean age at the time of the reconstruction was forty-nine years (range, twenty-two to seventy-three years), and the mean duration of follow-up was fifty-seven months (range, twenty-seven to 108 months). In five hips, bone-grafting of the acetabulum was needed to obtain superolateral coverage. The clinical result was excellent in eighteen hips and good in one. No revisions have been performed to date. Two femoral components were possibly loose radiographically. One was associated with a definite loosening of the acetabular cup. In addition, one other cup was possibly loose. There was a 100 per cent rate of survival if only a revision procedure was considered as a failure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408139 TI - Retention and strength of silicone-rubber catheters. A report of five cases of retention and analysis of catheter strength. AB - Five patients had a silicone-rubber catheter that broke during attempted removal. Mechanical testing was performed to determine the tensile strength of silicone rubber as well as of polyvinyl chloride catheters. Silicone-rubber catheters were found to have a low tensile strength. The pull-out force of ten indwelling silicone-rubber catheters was also measured. The pullout force measured from a patient who had a cast was close to 40 per cent of the strength of the silicone rubber catheters. PMID- 8408140 TI - Healing and remodeling of articular incongruities in a rabbit fracture model. AB - Healing of 0.5 or 1.0-millimeter step-off defects associated with displaced intra articular fractures of the medial femoral condyle was examined in fifty-four adult New Zealand White rabbits. The rabbits were treated with either immobilization for three weeks, intermittent active motion, or continuous passive motion for seven days. At twelve weeks, the healing and remodeling of the step off defects were examined with use of contact-pressure maps on pressure-sensitive film, light microscopy (with hematoxylin and eosin or safranin-O staining), and scanning electron microscopy. Macroscopically, the sharp profile that had been present initially with both sizes of step-off defect had rounded off; however, there was less residual incongruity with the 0.5-millimeter step-offs than with the 1.0-millimeter step-offs. Among step-off defects of the same size, the method of treatment had no discernible effect on the macroscopic appearance of the surface of the joint. With fresh step-offs (the control group), the contact pressure of the cartilage on the elevated side was approximately three times greater than that at a distance from the step-off. On the depressed side, an unloaded zone extended approximately three times the height of the step-off, with an average width of 3.4 millimeters for the 1.0-millimeter step-offs and 1.6 millimeters for the 0.5-millimeter step-offs. After healing and remodeling, the unloaded zone still averaged 2.5 millimeters in width for the 1.0-millimeter step offs but had decreased to only 0.35 millimeter in width for the 0.5-millimeter step-offs. For seven of the nine 0.5-millimeter step-offs, the contact pressure in the previously unloaded zone ranged from 0.5 to 1.5 megapascals, with a mean of 0.8 megapascal (40 per cent of the normal mean contact pressure at this location). Under light microscopy, the cartilage on the elevated side of the healed step-offs had decreased in thickness, was displaced toward the defect and tapered toward the depressed side, and ended in a hypocellular tissue flap. In contrast, the cartilage on the depressed side had thickened as a result of hyperplasia of the chondrocytes and hypertrophy of the cartilage and had failed to establish continuity between the sides of the defect. There was a marked increase in the subchondral vascular bed and re-establishment of the subchondral plate. With the exception of the aforementioned hypocellular tissue flap, safranin O stained the cartilage on both levels of the step-off uniformly, which indicated the absence of glycosaminoglycan depletion.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408141 TI - Recurrent dislocation of a hip with a labral lesion: treatment with a modified Bankart-type repair. Case report. PMID- 8408142 TI - Post-fracture lipid inclusion cyst. A case report. PMID- 8408143 TI - Soft-tissue aging and musculoskeletal function. PMID- 8408144 TI - Injury of the femoral artery in total hip replacement causing abdominal pain and hypovolemic shock. PMID- 8408146 TI - Extrapelvic compression of the sciatic nerve. An unusual cause of pain about the hip. PMID- 8408145 TI - Transverse carpal ligament. An important component of the digital flexor pulley system. PMID- 8408147 TI - Psychomotor education: point and counterpoint. PMID- 8408148 TI - Carpal tunnel release. A prospective, randomized assessment of open and endoscopic methods. AB - To define the role of two-portal endoscopic carpal-tunnel release as a method for the treatment of compression of the median nerve at the wrist, a prospective, randomized, multicenter study was performed on 169 hands in 145 patients. Either open or endoscopic carpal-tunnel release was performed in all of the patients who had clinical signs and symptoms consistent with carpal tunnel syndrome, had not responded to or had refused non-operative management, and had had electrodiagnostic studies consistent with carpal tunnel syndrome. Follow-up evaluations were performed at twenty-one, forty-two, and eighty-four days. At the end of the follow-up period, both the open and endoscopic methods had resulted in high levels of achievement of the primary outcomes (relief of pain and paresthesias). The numbness and paresthesias were relieved in eighty (98 per cent) of eighty-two hands in the open-release group compared with seventy-seven (99 per cent) of seventy-eight hands in the endoscopic-release group. This parameter was not recorded for three hands in the open-release group or six hands in the endoscopic-release group. The satisfaction of the patients with the procedure, graded on a scale of 0 to 100 per cent, averaged 84 per cent in the open-release group compared with 89 per cent in the group that had had endoscopic release. We found no significant differences between the two groups with regard to the secondary quantitative-outcome measurements, including two-point discrimination, postoperative interstitial-pressure data for the carpal canal, Semmes-Weinstein monofilament testing, and motor strength. The open technique resulted in more tenderness of the scar than did the endoscopic method. Thirty two (39 per cent) of eighty-two hands in the open-release group and fifty (64 per cent) of seventy-eight hands in the endoscopic-release group were not tender at eighty-four days. This parameter was not recorded for three hands in the open release group and six hands in the endoscopic-release group. The open method also resulted in a longer interval until the patient could return to work (median, twenty-eight days, compared with fourteen days for the open-release and endoscopic-release groups). Four complications occurred in the endoscopic carpal tunnel release group: one partial transection of the superficial palmar arch, one digital-nerve contusion, one ulnar-nerve neuropraxia, and one wound hematoma.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408149 TI - Skeletal metastases of unknown origin. A prospective study of a diagnostic strategy. AB - We carried out a prospective study of the effectiveness of a diagnostic strategy in forty consecutively seen patients who had skeletal metastases of unknown origin. The diagnostic strategy consisted of the recording of a medical history; physical examination; routine laboratory analysis; plain radiography of the involved bone and the chest; whole-body technetium-99m-phosphonate bone scintigraphy; and computed tomography of the chest, abdomen, and pelvis. After this evaluation, a biopsy of the most accessible osseous lesion was done. The laboratory values were non-specific in all patients. The history and physical examination revealed the occult primary site of the malignant tumor in three patients (8 per cent): one patient who had carcinoma of the breast; one, of the kidney; and one, of the bladder. Plain radiographs of the chest established the diagnosis of carcinoma of the lung in seventeen patients (43 per cent). Computed tomography of the chest identified an additional six primary carcinomas of the lung (15 per cent). Computed tomography of the abdomen and pelvis established the diagnosis in five patients (13 per cent): three patients who had carcinoma of the kidney; one, carcinoma of the liver; and one, carcinoma of the colon. Examination of the biopsy tissue established the diagnosis in only three additional patients (8 per cent) and confirmed it in eleven others. On the basis of the biopsy alone, we were unable to identify the primary site of the malignant tumor in twenty-six (65 per cent) of the patients. In thirty-four (85 per cent) of the forty patients, the primary site was identified with the use of the diagnostic strategy described here, and only two additional occult malignant tumors were found on follow-up studies. Our diagnostic strategy was simple and highly successful for the identification of the site of an occult malignant tumor before biopsy in patients who had skeletal metastases of unknown origin. PMID- 8408150 TI - Rheumatoid arthritis of the cervical spine. A long-term analysis with predictors of paralysis and recovery. AB - We analyzed the cases of seventy-three patients who were managed over a twenty year period for rheumatoid involvement of the cervical spine and were followed for a minimum of two years, with an average follow-up of seven years. A neurological deficit did not develop in thirty-one patients (Ranawat et al. Class I) and paralysis developed in the remaining forty-two patients: Class II in eleven and Class III in thirty-one. Of the forty-two patients in whom paralysis developed, thirty-five had operative stabilization. Seven patients were managed with a soft cervical collar because they refused or were medically unable to have the operation; all of the had an increase in the severity of the paralysis. The posterior atlanto-odontoid interval and the diameter of the subaxial sagittal canal measured on the cervical radiographs demonstrated statistically significant correlations with the presence and severity of paralysis. All of the patients who had a Class-III neurological deficit had a posterior atlanto-odontoid interval or diameter of the subaxial canal that was less than fourteen millimeters. In contrast, the anterior atlanto-odontoid interval, which has traditionally been reported, did not correlate with paralysis. The prognosis for neurological recovery following the operation was not affected by the duration of the paralysis but was influenced by the severity of the paralysis at the time of the operation. The most important predictor of the potential for neurological recovery after the operation was the preoperative posterior atlanto-odontoid interval. In patients who had paralysis due to atlanto-axial subluxation, no recovery occurred if the posterior atlanto-odontoid interval was less than ten millimeters, whereas recovery of at least one neurological class always occurred when the posterior atlanto-odontoid interval was at least ten millimeters. If basilar invagination was superimposed, clinically important neurological recovery occurred only when the posterior atlanto-odontoid interval was at least thirteen millimeters. All patients who had paralysis and a posterior atlanto-odontoid interval or diameter of the subaxial canal of fourteen millimeters had complete motor recovery after the operation. In this series, although only patients who had a neurological deficit were operated on, we observed the range of the posterior atlanto-odontoid interval that was associated with poor or no recovery after the operation, and we identified the safe range on the basis of the patients in whom paralysis did not develop.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408151 TI - Robinson anterior cervical discectomy and arthrodesis for cervical radiculopathy. Long-term follow-up of one hundred and twenty-two patients. AB - We evaluated the results of the Robinson method of anterior cervical discectomy and arthrodesis with use of autogenous iliac-crest bone graft, at one to four levels, in 122 patients who had cervical radiculopathy. A one-level procedure was done in sixty-two of the 122 patients; a two-level procedure, in forty-eight; a three-level procedure, in eleven; and a four-level procedure, in one. The average duration of clinical and roentgenographic follow-up was six years (range, two to fifteen years). The average age was fifty years (range, twenty-five to seventy eight years). Preoperatively, 118 patients had pain in the arm, fifty-five had weakness of one or more motor roots, and seventy-seven had sensory loss. At the time of follow-up, eighty-one patients had no pain in the neck, twenty-six had mild pain in the neck, nine had moderate pain in the neck, four had mild radicular pain, and two had a combination of mild radicular pain and moderate pain in the neck. One hundred and eight patients had no functional impairment, and fourteen had a slight limitation of function during the activities of daily living. Nine of eleven patients who had symptoms related to a change at one level cephalad or caudad to the site of a previous arthrodesis had another operative procedure. Lateral roentgenograms of the cervical spine, made in flexion and extension, showed a pseudarthrosis at twenty-four of 195 operatively treated segments. Sixteen of the patients who had a pseudarthrosis were symptomatic, but only four had sufficient pain to warrant revision. The risk of pseudarthrosis was significantly greater after a multiple-level arthrodesis than after a single level arthrodesis (p < 0.01). At the time of the most recent follow-up, fifty three of the fifty-five patients who had had a motor deficit had had a complete recovery, and the two remaining patients had had a partial recovery. Seventy-one of the seventy-seven patients who had had a sensory loss had regained sensation. None of the patients had an increased neurological deficit postoperatively. Our results suggest that the Robinson anterior cervical discectomy and arthrodesis with an autogenous iliac-crest bone graft for cervical radiculopathy is a safe procedure that can relieve pain and lead to resolution of neurological deficits in a high percentage of patients. PMID- 8408152 TI - Evaluation and treatment of diastematomyelia. AB - We reviewed the results for forty-three patients who had a diastematomyelia. All of the patients had been skeletally immature when the diagnosis was made, the mean age being six years (range, birth to thirteen years), and were skeletally mature by the time that they were evaluated by us. When they were first seen at our institution, twenty-four patients (56 per cent) had a cutaneous lesion, such as hairy patch, dimple, hemangioma, subcutaneous mass, or teratoma at or near the level of the diastematomyelia; thirty-four patients (79 per cent) had congenital scoliosis; and forty-two patients (98 per cent) had at least one associated musculoskeletal anomaly, such as spinal dysraphism, asymmetry of the lower extremities, club foot, or a cavus foot. In twenty-seven patients (63 per cent), the diastematomyelia was located in the lumbar spine. Thirty-six patients had eighty-four neurological manifestations. Resection of the spur was performed in thirty-three patients at a mean age of seven years (range, three months to seventeen years). Twenty-two patients who had a resection had no change in neurological condition, nine patients had improvement, and one patient had one symptom improve and another symptom worsen after the operation. We believe that resection of the spur should be performed in patients who have progressive neurological manifestations. Patients who do not have progressive neurological manifestations should be observed; if progression is noted, a resection should then be performed. PMID- 8408153 TI - Determination of acetabular coverage of the femoral head with use of a single anteroposterior radiograph. A new computerized technique. AB - We developed a method for the estimation of three-dimensional acetabular coverage of the femoral head with use of only an anteroposterior radiograph of the hip. This technique also allows recalculation of the corrected value for coverage at neutral pelvic tilt. Provided that the acetabulum and femoral head are spherical and congruent, the results are as accurate as those obtained with computerized tomographic reconstruction, the dose of radiation is much lower, and much less time is required for calculation. The hips of 286 normal subjects showed increases with age in both anterior and posterior coverage and backward tilt of the pelvis, along with a decrease in the anterior-posterior ratio of coverage. The proportion of anterior acetabular coverage in female subjects was smaller than that in male subjects. There was more backward tilt of the pelvis and the anterior-posterior ratio of coverage was smaller when the subjects were standing than when they were supine. PMID- 8408154 TI - Open reduction through a medial approach for congenital dislocation of the hip. A critical review of the Ludloff approach in sixty-six hips. AB - We reviewed the results of open reduction through the medial approach of Ludloff, done for congenital dislocation in sixty-six hips (sixty-three children). The mean age at the time of the operation was twelve months (range, two to sixty three months), and the mean duration of follow-up was six years (range, two to thirteen years). Avascular necrosis was evident preoperatively in two hips (3 per cent) and was noted postoperatively in another seven hips (11 per cent). There was a correlation between the age of the patient at the time of the operation and postoperative avascular necrosis, with an increased prevalence of the complication in patients who had been managed with the open reduction after the age of twenty-four months. One redislocation and two subluxations were noted at the time of the first changing of the cast, four weeks postoperatively. Although the acetabular index decreased from a mean of 38 degrees preoperatively to a mean of 16 degrees at the time of follow-up, acetabular dysplasia did not resolve in 33 per cent of the hips and pelvic osteotomy was performed. We consider the Ludloff approach to be a safe and effective method for the treatment of congenital dislocation of the hip in infants who are less than the age of twenty four months and in whom a concentric reduction with less than 60 degrees of abduction was not achieved following closed reduction. The advantages of this approach include direct access to the iliopsoas, the transverse acetabular ligament, and the constricted capsule; minimum loss of blood; and a cosmetically acceptable scar. PMID- 8408155 TI - Arthroscopically assisted reconstruction of the anterior cruciate ligament with use of autogenous patellar-ligament grafts. Results after twenty-four to forty two months. AB - The results of the first sixty-nine consecutive patients who had had seventy arthroscopically assisted reconstructions of the anterior cruciate ligament with use of an autogenous patellar-ligament graft at our institution were reviewed retrospectively. Sixty-seven patients (sixty-eight knees) were available for evaluation after a minimum of two years. All patients had been managed with early, postoperative range-of-motion exercises and a standardized program of physical therapy. At the time of the most recent follow-up evaluation, the median ligament score, according to the rating system of The Hospital for Special Surgery, was 93 of a possible 100 points. Of the sixty-eight knees, forty-four were rated excellent; fifteen, good; six, fair; and three, poor. Eighteen knees had symptoms related to the patellofemoral joint and sixty-three had a full range of motion; two knees had had manipulation for loss of flexion. At the follow-up evaluation, KT-1000 arthrometric measurements were obtained for both knees of fifty-six patients. Eighty-four per cent of the patients had an increase of three millimeters or less in anterior-posterior displacement of the tibia on the reconstructed side compared with the normal side, while 93 per cent had an increase of four millimeters or less. Postoperatively, there was no apparent association between changes in the Insall-Salvati patellar ligament-to-patella ratios and pain in the patellofemoral joint. The results of the arthroscopically assisted reconstructions combined with use of early range-of-motion exercises were comparable with those reported after open reconstruction and immobilization of the limb in a plaster cast. The frequency of pain in the patellofemoral joint and the need for manipulation because of loss of motion were decreased after the arthroscopically assisted procedures. PMID- 8408156 TI - Reduction of postoperative blood loss after press-fit condylar knee arthroplasty with use of a femoral intramedullary plug. AB - In a prospective study of eighty press-fit condylar knee arthroplasties performed with cement, we compared the postoperative blood loss in forty patients in whom the defect left by the femoral intramedullary alignment rod had been left open with the blood loss in forty patients in whom the defect had been closed with a plug of cement. The patients in the open-defect group had significantly more blood loss (mean, 1002 milliliters) in the first twenty-four hours after the operation than the closed-defect group (mean, 752 milliliters) (p < or = 0.01). The total postoperative blood loss was also significantly greater in the open defect group (mean, 1536 milliliters) than in the closed-defect group (mean, 1215 milliliters) (p = 0.02). PMID- 8408157 TI - Mechanical consequences of core drilling and bone-grafting on osteonecrosis of the femoral head. AB - We employed an anatomically realistic three-dimensional finite-element model to explore several biomechanical variables involved in coring or bone-grafting of a segmentally necrotic femoral head. The mechanical efficacy of several variants of these procedures was indexed in terms of their alteration of the stress:strength ratio in at-risk necrotic cancellous bone. For coring alone, the associated structural compromise was generally modest, provided that the tract did not extend near the subchondral plate. Cortical bone-grafting was potentially of great structural benefit for femoral heads in which the graft penetrated deeply into the superocentral or lateral aspect of the lesion, ideally with abutment against the subchondral plate. By contrast, central or lateral grafts that stopped well short of the subchondral plate were contraindicated biomechanically because they caused marked elevations in stress on the necrotic cancellous bone. Calculated levels of stress were relatively insensitive to variations in the diameter of the graft. PMID- 8408158 TI - Thoracic outlet syndrome associated with an anomalous coracoclavicular joint. A case report. PMID- 8408159 TI - Posterior dislocation of the humeral head in infancy associated with obstetrical paralysis. A case report. PMID- 8408160 TI - Injuries of the posterior cruciate ligament. PMID- 8408161 TI - Geographic variations in the rates of elective total hip and knee arthroplasties among Medicare beneficiaries in the United States. PMID- 8408162 TI - Pulmonary embolism after total hip replacement in a patient who had an ovarian cyst. A case report. PMID- 8408163 TI - Biologic response modifier OK-432 pretreatment reduces mortality in burned mice infected with gram-negative bacteria. AB - When compared with a control group, mice pretreated with the biologic response modifier OK-432 showed increased survival when burned and infected with several strains of Pseudomonas aeruginosa and a Klebsiella pneumoniae isolate. Quantitative microbial counts in the skin, liver, and kidneys of burned mice infected with Pseudomonas aeruginosa showed that although skin counts were not different between control groups pretreated or untreated with OK-432, liver and kidney counts were reduced significantly in the group pretreated with OK-432. We speculate from these results that OK-432 pretreatment may enhance survival by enhancing host clearance of microorganisms entering the circulation from the burn wound. Making mice granulocytopenic by cyclophosphamide treatment or macrophage deficient/dysfunctional by carrageenan treatment nullified any survival advantage given by OK-432 pretreatment. These results supported our speculation and suggested that both of these cell types were important in the mechanisms of enhanced survival afforded by OK-432 pretreatment. PMID- 8408164 TI - Diaphragm acetylcholinesterase multimeric forms in mice in response to burn trauma. AB - This report continues our investigations concerning relative changes in acetylcholinesterase (AChE) activity in response to burn trauma. This experiment was performed in mouse diaphragm. AChE is fundamental to the operating efficiency of the neuromuscular junction. It is through such junctions that all skeletal muscle contraction occurs. This paper tests the hypothesis that burn trauma induces changes in diaphragm AChE with respect to total content, specific molecular forms, and relationships of the polymorphic forms to each other. After mice were exposed to increasing size of body surface area (20%, 30%, and 50%) diaphragms were excised at 3 weeks after burn injury. AChE activity was measured by differential extraction and velocity sedimentation on sucrose gradients. In the sham-treated control groups most of the AChE activity consisted entirely (92%) of the globular forms (S1 and S2). Asymmetric (S3 and S4) and nonextractable (H5) forms made up 2% and 6% of the AChE activity, respectively. In the burn groups all total globular activity was decreased by 1/6 to 1/3 of control groups. For these groups the asymmetric forms (S3 and S4) were increased ninefold to fourteenfold. Nonextractable forms (S5) among the groups were not different. Individual soluble globular forms (S1: 4S, 6.5S, 10S) were reduced by 50% for only the 20% and 30% BSA groups. Residual globular forms (S2) were increased threefold to ninefold. Some of the asymmetric forms (S3: 4S, 10S, 12S, and 16S) were increased fivefold to eightfold among the burn groups. Residual asymmetric (S4) and nonextractable (S5) forms were not different. This report shows that in response to burn trauma the AChE activity of the diaphragm and the interdependence of the polymorphic forms were changed at the chronic postburn period. These findings may aid in understanding the adaptive response of diaphragmatic preservation in burn trauma. PMID- 8408165 TI - Relationships between heat production, heat loss, and body temperature for rats with burn injuries between 26% and 63% of the body surface area. AB - Burn injury in man is characterized by increased body temperature proportional to burn wound size and may represent fever and/or hyperthermia. A nonseptic animal model used to study this phenomena has not been described. To test the hypothesis that large burn injuries in rats would produce increased body temperature, the rectal, skin, and body temperatures were sequentially measured and were calculated for rats in the control group and rats with burn injuries covering 26% to 63% of the body surface area [< or = 35%, 36% to 45%, and > or = 46% body surface area]. The group with burns covering > or = 46% of the body surface area had significantly higher rectal temperatures than did at least one other group on postburn days 7, 9, 11, 13, 18, and 20. On postburn days 7 and 11 this increase was significantly higher than that of all burn and control groups. Animals did not demonstrate any overt evidence of wound infection. These data do not establish a cause for increased body temperature after burn injury but suggest that a reproducible animal model may be possible for the study of the cause of increased body temperature after burn injury. PMID- 8408166 TI - Temperature changes during exercise stress testing in children with burns. AB - It has been postulated that because of the extensive destruction of the skin and appendages after thermal injury, the thermoregulatory control mechanism would be impaired, and these patients would be intolerant to prolonged work. Preview studies demonstrate evidence that during work in a hot climate, patients with an extensively healed burn react with an excessive rise in body temperature. This study was designed to investigate the thermoregulatory response to exercise in pediatric patients with burns and to study changes in body temperature during exercise testing. Cardiopulmonary stress tests were completed in 32 children with a mean postburn time of 2.3 +/- 1.5 years and a mean burn size of 44% +/- 23% total body surface area. Exercise variables included expired volume, tidal volume, respiratory rate, tidal/dead space rate, heart rate, and work stage achieved. Temperature monitoring included external auditory canal temperature, burn scar, and normal skin temperature. Values were measured at baseline during and at maximum exercise. Our data indicate all patients reached the same endurance level regardless of the size of the total body surface area burn. Additionally, in a temperature-controlled environment, adequate heat dissipation in children with burns can be maintained during exercise testing without an excessive rise in body temperature. PMID- 8408167 TI - Enhancement of the burned nasal tip with the use of upper lip scar flaps. AB - Maximal aesthetic improvement for the patient with burns deserves the same considerations as does functional restoration. The use of burn scars as vascularized flaps is a prime example in which these goals may be simultaneously achieved with an added bonus of salvaging tissues that would otherwise be discarded. Several cases involving revision of commonly encountered perioral scars demonstrate the effectiveness of burn scar flaps in the correction of concomitant nasal deformities, and thus reemphasize this useful principle. PMID- 8408168 TI - A simplified testing system to evaluate performance after transplantation of human skin preserved in glycerol or in liquid nitrogen. AB - We have designed and tested a mouse recipient model for evaluation and direct comparison of skin-preservation procedures. Glycerolized skin was compared with cryopreserved and fresh cadaveric skin. Human skin samples were grafted on Balb/c mice, and primary take was evaluated after 4 and 7 days. The results demonstrate that although all grafted specimens were initially accepted as indicated by gross observations, histologic differences were evident and significant. Cryopreserved skin grafts performed better than did glycerolized skin even after a transplantation period as short as 4 days; this difference became even more pronounced after 7 days. Both methods of preservation provided a less successful product than did fresh viable cadaveric skin. However, for very short periods of grafting the performance of glycerolized skin might be considered adequate (79% compared with cryopreserved). Because the present study used an immunocompetent xenogeneic model, it is possible that the period of adequate allogeneic grafting will be prolonged in patients who are immunosuppressed. The present model is simple to use, requires a minimum of maintenance and expertise, and has inherent clinical relevance, because it is concerned primarily with the clinical performance of the skin. Thus it may be used as a quality control test for banked skin. PMID- 8408169 TI - Severe multiple mononeuropathy in patients with major thermal burns. AB - Multiple mononeuropathy after thermal burns covering greater than 40% total body surface area occurred in nine of 121 of our burn center admissions for an incidence of 7.4%. The number of nerves involved per patient ranged from three to seven, with the average being 4.9. Upper-extremity nerves were more commonly involved than were lower extremity nerves (33 versus 11). All patients had burns over the involved areas. The source was believed to be due to a multiple crush syndrome, in which multiple different neuropathic factors in each patient summate to cause a multiple mononeuropathy. The outcome with conservative treatment was variable, with lower-extremity nerve lesions having a very good outcome and upper extremity lesions not having as good an outcome, particularly when the median nerve was affected. PMID- 8408170 TI - Anesthesia-assisted procedures in a burn intensive care unit procedure room: benefits and complications. AB - A retrospective review of 109 procedures was performed to evaluate the safety and efficacy of anesthesiologist-administered anesthesia in the burn intensive care unit treatment room. Intraprocedural and postprocedural complications, impact on patient activity, and nutritional goals were evaluated. The review suggested that these procedures can be performed safely with appropriate supervision and monitoring without detrimental effects on patient activity level or nutritional status. PMID- 8408171 TI - Evaporative water losses through a temporary wound dressing under simulated wound conditions. AB - Patients with burns lose large amounts of water through evaporation from open wounds. Because the wound covering is the first line of defense for maintenance of body fluid balance in these patients, quantification of the evaporative water loss through wound coverings at the bedside would improve the accuracy of estimations of body water loss. The present experiment evaluates the use of a small ventilated capsule system automated with miniature resistance-type dew point sensors for measurement of evaporative water loss through biologic dressings under simulated wound conditions. Evaporative water loss from wounds was simulated by pilocarpine-induced profuse sweating on the forearm. Evaporative water loss through uncovered skin was compared with that of skin covered with commercially available temporary wound dressings. Compared with an adjacent unstimulated area, forearm dew-point temperature in the capsule (Tcdp) and sweat rate increased immediately after pilocarpine exposure and remained significantly elevated and relatively constant for an additional 60 minutes. Evaporative water loss of the forearm was 29 +/- 4.8 gm/m2/hr (mean +/- SE) at baseline and rose significantly to 275 +/- 18.2 gm/m2/hr after pilocarpine exposure. The pilocarpine-stimulated sweat rate and Tcdp at neutral conditions were similar to those obtained from walking on a treadmill for 60 minutes in a 30 degrees C room. Compared with pilocarpine-induced evaporative water loss of the uncovered skin, temporary wound dressings significantly reduced evaporative water loss by 40% to 60%. No significant differences were observed between varieties of temporary wound dressings differing in thickness and/or porosity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408172 TI - Outpatient skin grafting of extremity burn wounds with the use of Unna Boot compression dressings. AB - Thirty-one patients underwent split-thickness skin grafting for burn injuries of an extremity, after which Unna Boot compression dressings were applied for fixation of the graft. Three patients required hospitalization of 2 to 4 days, and 28 patients were treated strictly on an outpatient basis. The lower extremity was involved in 25 patients, and the upper extremity was involved in six. Average wound size was 284 cm2. Eighteen patients were treated with sheet grafts, and 13 received meshed grafts. Nine wounds extended across a joint. Patients were allowed immediate ambulation after surgery. All grafts resulted in 95% to 100% wound coverage, and no regrafting was required. Application of Unna boot compression dressings to extremity skin grafts provides excellent protection of both meshed and nonmeshed grafts and allows immediate ambulation and range of motion. Many patients with burn injuries may be treated on an outpatient basis with the use of this technique. PMID- 8408173 TI - Effect of triglycyl-lysine-vasopressin on skin blood flow and blood loss during wound excision in patients with burns. AB - Excisional therapy often results in large-volume blood loss. Triglycyl-lysine vasopressin selectively decreases dermal blood flow and therefore was tested for efficacy in limiting intraoperative blood loss in a series of patients undergoing excisional therapy. Ten patients with symmetric injuries were treated with intravenous triglycyl-lysine-vasopressin after excision of half of their burn wound. Blood loss, which was quantified by weighing sponges used to absorb shed blood, was significantly decreased after treatment. Triglycyl-lysine-vasopressin treatment was safe and effective and should be considered in cases when large volume blood loss is expected. PMID- 8408174 TI - Trichloroethylene "burn". PMID- 8408175 TI - Stapled tie-over stent: a simplified technique for pressure dressings on newly applied split-thickness skin grafts. AB - A simplified technique for simultaneously securing a skin graft and applying a stented pressure dressing is described. This method provides good fixation and immobilization of the graft. It is less time-consuming than previously described methods, and it is easier to perform. PMID- 8408176 TI - The use of the modified Thomas splint for a patient with cultured epidermal autografts. AB - Because of the difficulty in handling a patient with cultured epidermal autografts in pressure areas of the limbs, we have adopted a splint that is easy to apply, allows an easy access to the wound, and is well tolerated by the patient. HYPOTHESIS: Avoiding pressure in areas with autografts is recommended. This is the reason we have used a splint that avoids the pressure and the secretion buildup. This technique helps a better "take of the grafts." METHODS: We have modified the original Thomas splint in two ways: (1) the use of a distant and upper appliance that allows to correct the foot's flexion; and (2) the use of an appliance that can be regulated, is inflatable, and can be removed, allowing the change of pressure in different areas. We have used this method with one patient with a deep degree of circumferential burn in both lower limbs. The splint has been maintained and the elevation changed every day for 15 days. Traction has been made with the use of the Kirschner stirrup. RESULTS: The dressing was changed easily and without patient pain. Good mobility was possible, with genital hygiene and grafts that were taken in 70% of the cultured epidermal autografts. CONCLUSION: This method allows maintenance of the legs in functional position during dressing changes and avoids pressure and chafing. It decreases the loss of the autograft for mechanical effects. PMID- 8408177 TI - The status of statewide burn prevention legislation. AB - Successful programs in injury prevention can focus on changing an individual's attitude and behavior or on creating and amending the environment to reduce the likelihood of injury. To investigate the latter approach as it pertains to burn prevention, the American Burn Association Burn Prevention Committee catalogued statewide legislation (as opposted to county or municipal ordinances) in major areas of burn and fire prevention. Burn reporting was generally viewed by state fire marshals as a means to apprehend arsonists or investigate child abuse, as opposed to collect demographic data for injury prevention. Smoke detectors are mandated in new residential construction by the majority of states; however, one in six still leave this to local initiatives. Sprinkler systems have generally not been addressed by state legislation. The committee concludes that pursuing statewide legislative agendas as an area of burn and injury prevention is open to further initiatives. PMID- 8408178 TI - Self-inflicted burns. AB - Over a 3-year period 17 people were admitted for intentionally self-inflicted burns. The mean total body surface area burn was 29.5%, and 59% of the patients sustained an inhalation injury. Two patients died from their injuries (one male and one female). The method most often used (59%) was a flammable liquid ignited by a flame. Of those patients, 50% used gasoline. Fifty-nine percent of the patients were current substance abusers, with alcohol (80%) being the favored drug. Aside from substance abuse, psychiatric abnormalities were present in 53% of the patients. Schizophrenia/schizoaffective disorders was the primary psychiatric diagnosis (44%), with most patients having undergone previous psychiatric treatment. All had diagnosed disorders (using the Diagnostic and Statistical Manual III--Revised) including substance abuse and/or other psychiatric disorders. Actual stated suicidal intent was present in 41% of the cases, and another 41% were irrational attempts to escape from or control emotional pain. PMID- 8408179 TI - A psychiatric perspective on self-inflicted burns. PMID- 8408180 TI - Protein kinase C and adenylate cyclase as targets for growth inhibition of human gastric cancer cells. AB - In the human gastric adenocarcinoma cell line AGS the effects of the protein kinase-C-activating phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA), the protein kinase C inhibitor staurosporine, the adenylate-cyclase activating agent forskolin, and the permeable dibutyryl-adenosine 3',5'-monophosphate (Bt2cAMP) on the proliferation were assessed. Cell counting followed 5 days of incubation. Prolonged activation of protein kinase C by TPA, inhibition of protein kinase C by staurosporine, activation of adenylate cyclase by forskolin or a direct increase of the intracellular cAMP level all result in a dose-dependent growth inhibition of AGS gastric tumour cells. Half-maximal inhibition was achieved at 100 pM for TPA, 1 nM for staurosporine, 20 microM for forskolin, and 600 microM for Bt2cAMP. It is concluded that protein kinase C and adenylate cyclase play a fundamental role in the growth of AGS gastric cancer cells. Interference with these enzymes involved in the signal transduction of growth regulation in tumour cells may represent a target in the development of new antiproliferative principles. PMID- 8408181 TI - Why does a higher response rate to chemotherapy correlate poorly with improved survival? PMID- 8408182 TI - Chemotherapeutic effect on osteosarcoma on basis of collagen analysis: a proposal of the induction of osteosarcoma differentiation. AB - The authors studied effect of chemotherapy on osteosarcoma by collagen analysis. As a result of this case study we propose the induction of osteosarcoma differentiation by chemotherapy. Treatment of a conventional osteosarcoma with two intra-arterial infusions of cisplatin and the T-12 protocol of Rosen resulted in sclerotic changes and good margination accompanied by the disappearance of the soft-tissue component from the X-rays. More than 90% tumour destruction was histologically demonstrated; tumour bone and osteoid increased after the chemotherapy, and the viable area of the tumour resembled an osteoblastoma. Before the chemotherapy, immunolocalization determined collagen types I and V to be diffusely present in the bone and osteoid. After the chemotherapy, the antibody to type I collagen was diffusely present, but the antibody to type V collagen occurred only on the surface of the increased bone and osteoid as in normal bone. When osteosarcoma cells were treated in vitro with methotrexate or cisplatin, collagen production increased significantly. It is thus believed that tumour cells were directly stimulated with these chemotherapeutic agents to produce collagen. The findings suggested that some anticancer agents might not only be cytotoxic to but also differentiate osteosarcoma cells. PMID- 8408183 TI - [meso-1,2-bis(2,6-dichloro-4-hydroxyphenyl)ethylenediamine]- dichloroplatinum(II), a new drug not only parenterally but also orally active in the therapy of breast and prostate cancer. AB - The platinum complex [meso-1,2-bis(2,6-dichloro-4-hydroxyphenyl)- ethylenediamine]dichloroplatinum(II),K, was tested for its antitumor activity on hormone-sensitive tumor models under peroral administration. The resorption from the gastrointestinal tract was proved by determining the estrogenic effect of K in a dose/activity study using the immature-mouse uterine weight test. In comparison to the subcutaneous injection, a tenfold peroral dose was administered to achieve identical effects. By peroral treatment of the hormone-sensitive MXT(M3.2) mammary carcinoma of the mouse with K an almost complete inhibition of the tumor growth was obtained. This effect was superior to that of subcutaneously applied cisplatin and significantly better than that obtained by perorally administered ligand L at an equimolar dose, indicating that the antitumor effect is caused by the intact complex K and not by the liberated ligand L. The strong antitumor activity of perorally applied K was also demonstrated on the hormone sensitive Noble Nb-R prostatic carcinoma of the rat. Histological examinations showed that the platinum complex K did not cause cisplatin-like kidney damage or irritations of gastric or intestinal mucosa when given perorally. PMID- 8408184 TI - Esophageal carcinoma in house musk shrews, Suncus murinus (Insectivora), induced by N-methyl-N'-nitro-N-nitrosoguanidine. AB - Female 6-week-old shrews were given a solution of N-methyl-N'-nitro-N nitrosoguanidine (MNNG) at a concentration of 50 micrograms/ml or 100 micrograms/ml in the drinking water. All 11 shrews receiving 100 micrograms/ml MNNG died 8-13 days after the beginning of carcinogen administration and 6 of the 20 shrews receiving 50 micrograms/ml MNNG died after 10-54 days. When animals were between 43 and 54 weeks of age, multiple esophageal lesions were evoked in all 14 that had received 50 micrograms/ml MNNG for 30 weeks. All shrews developed a protruding, ulcerative, or superficial type of squamous-cell carcinoma of the esophagus, accompanied by papillomas. Local invasion was seen in squamous-cell carcinoma but no distant metastasis was noted. None of the 5 control shrews developed any esophageal abnormality. No gastric adenocarcinoma, intestinal sarcoma, or other tumors were induced with MNNG. It can be concluded that MNNG has a carcinogenic effect on shrew esophageal epithelium. PMID- 8408185 TI - In vitro studies on interaction of 4-hydroperoxyifosfamide and 2 mercaptoethanesulphonate in malignant gliomas. AB - Drug interference of ifosfamide and sodium 2-mercaptoethanesulphonate (MESNA) was studied in three malignant glioma cell cultures (HTZ-17, HTZ-209B, and HTZ-243) by a recently developed in vitro method for evaluation of multimodal treatment interactions. Glioma cell cultures were treated in monolayer 96-well tissue culture plates for 2 h each, with 4-hydroperoxyifosfamide and MESNA combined in both sequences, or alone. Concentrations ranged from 0.01 microM to 50 microM in single-modality exposures, and from 0.01 microM to 10 microM in combination exposures. After five population doubling times, DNA synthesis was determined by a standard [3H]Tdr-incorporation liquid-scintillation-counting protocol. Data points were evaluated for mono- and combined treatment dose effects (adapted with a probit function), and a model-free three-dimensional response surface was created that was compared to the theoretical additive, anticipated response surface. Local additivity was analysed for any ratio of combined treatment. No tumour effects were seen with MESNA in single-drug exposure, whereas ifosfamide resulted in more than 90% inhibition of tumour DNA synthesis. In combination experiments, MESNA could be confirmed to be inert: the anticipated theoretical combination response surfaces formed a three-dimensional extension of the single drug ifosfamide dose/response curves--the experimental combination response surfaces displayed an identical appearance (P < or = 0.05). In conclusion, these results indicate no drug interference of MESNA and ifosfamide in malignant glioma cells. PMID- 8408186 TI - A specific and rapid receptor assay for squamous-cell carcinoma of human esophagus. AB - A specific radioreceptor assay, for squamous-cell carcinoma of the human esophagus, using 2G3 monoclonal antibody, has been developed for the first time. In this assay a new mathematical parameter of the Scatchard plot has been introduced for the correct measure of binding capacity (BCcm = x cos alpha). This is the perpendicular distance from the origin to the Scatchard plot line. It is always a positive quantity and is directly proportional to both the x and y intercepts for expressing the binding capacity. The assay is highly sensitive and can be used to differentiate various types of esophageal tumors such as squamous cell carcinoma, adenocarcinoma, ulcerative growth and also a benign growth. BCcm significantly varies in the case of squamous-cell carcinoma tumors compared to adenocarcinoma and other types of tumor of the human esophagus. The assay can be completed in about 4 h and it may be used as a clinical diagnostic test. PMID- 8408187 TI - Renal, metabolic, and hemodynamic side-effects of interleukin-2 and/or interferon alpha: evidence of a risk/benefit advantage of subcutaneous therapy. AB - Systemic immunotherapy with recombinant interleukin-2 (rIL-2) via intravenous (i.v.) and subcutaneous (s.c.) administration produces objective responses in a proportion of advanced cancer patients. While most of the previous investigations chose the i.v. route for cytokine application, there is an increasing number of trials employing s.c. rIL-2 therapy. The comparison of reported response rates for i.v. versus s.c. therapy reveals no significant differences between these modalities. In an effort to describe systemic toxicities of s.c. cytokine therapy with regard to renal, metabolic, and hemodynamic abnormalities and to compare these effects to toxicities reported upon i.v. therapy, we retrospectively evaluated 148 treatment cycles of s.c. immunotherapy given to 107 outpatients. Our study cohorts consisted of 15 patients who received s.c. rIL-2 at doses of (4.8-14.4) x 10(6) IU m-2 day-1 5 days/week for a total of 8 weeks, 20 patients who received rIFN alpha 2b at (3.0-6.0) x 10(6) m-2 day-1 thrice weekly for a total of 6 weeks, and 72 patients who were given s.c. rILFN alpha 2b at 6.0 x 10(6) U/m2, three times per week, plus s.c. rIL-2 at (14.4-18.0) x 10(6) IU/m2 on days 1 and 2, followed by 4.8 x 10(6) IU m-2 day-1 5 days/week for 6 consecutive weeks. These treatment regimens were well tolerated in the outpatient setting; no toxic death occurred, and none of the patients developed life-threatening toxicity due to a capillary leak syndrome. Upon s.c. combination therapy, dyspnea at rest occurred in 6% of patients and grade III and IV hypotension occurred in 7% and 4%, respectively; plasma protein was significantly decreased (mean nadir +/- standard deviation, 67 +/- 5 g/l). In addition, s.c. therapy led to a significant increase in serum creatinine (mean peak +/- standard deviation, 115.1 +/- 21.4 mumol/l) and urea nitrogen (mean peak +/- standard deviation, 6.5 +/- 2.5 mmol/l); electrolyte disturbances and direct nephrotoxicity never caused major clinical symptoms. This was in marked contrast to a multitude of dose limiting and life-threatening adverse reactions reported upon i.v. rIL-2 therapy. We conclude that palliative low to intermediate-dose s.c. rIL-2/rIFN alpha combination therapy, in contrast to i.v. treatment, can be administered in the ambulatory setting with good practicability and excellent safety. This outpatient regimen is as effective against metastatic renal cell cancer as the most aggressive i.v. rIL-2 protocol reported. PMID- 8408188 TI - Chemotherapy for recurrent adeno- and adenoidcystic carcinomas in the head and neck. AB - We have investigated the effectiveness of chemotherapy for patients with recurrent adeno- and adenoidcystic carcinomas in the head and neck. Fourteen cases received a monthly combination chemotherapy regimen of cyclophosphamide, pirarubicin and cisplatin (CAP therapy). A response rate of 36% (5/14) was achieved. There was one complete response and four partial responses. The median duration of response was 37 months in the complete response case and 16 months (range, 6 to 20) in the partial response cases. The toxicity of this chemotherapy was acceptable. The result demonstrates that CAP therapy is an effective regimen for adeno- and adenoidcystic carcinomas. It may also be available as induction or adjuvant chemotherapy for patients with advanced tumors of these cancers. PMID- 8408189 TI - Report from the 24th annual meeting of the Society of Gynecologic Oncologists, February 7-10, 1993. PMID- 8408190 TI - Focusing on unpolymerized actin. PMID- 8408191 TI - A dual role for mitochondrial heat shock protein 70 in membrane translocation of preproteins. AB - The role of mitochondrial 70-kD heat shock protein (mt-hsp70) in protein translocation across both the outer and inner mitochondrial membranes was studied using two temperature-sensitive yeast mutants. The degree of polypeptide translocation into the matrix of mutant mitochondria was analyzed using a matrix targeted preprotein that was cleaved twice by the processing peptidase. A short amino-terminal segment of the preprotein (40-60 amino acids) was driven into the matrix by the membrane potential, independent of hsp70 function, allowing a single cleavage of the presequence. Artificial unfolding of the preprotein allowed complete translocation into the matrix in the case where mutant mt-hsp70 had detectable binding activity. However, in the mutant mitochondria in which binding to mt-hsp70 could not be detected the mature part of the preprotein was only translocated to the intermembrane space. We propose that mt-hsp70 fulfills a dual role in membrane translocation of preproteins. (a) Mt-hsp70 facilitates unfolding of the polypeptide chain for translocation across the mitochondrial membranes. (b) Binding of mt-hsp70 to the polypeptide chain is essential for driving the completion of transport of a matrix-targeted preprotein across the inner membrane. This second role is independent of the folding state of the preprotein, thus identifying mt-hsp70 as a genuine component of the inner membrane translocation machinery. Furthermore we determined the sites of the mutations and show that both a functional ATPase domain and ATP are needed for mt hsp70 to bind to the polypeptide chain and drive its translocation into the matrix. PMID- 8408192 TI - Presequence and mature part of preproteins strongly influence the dependence of mitochondrial protein import on heat shock protein 70 in the matrix. AB - To test the hypothesis that 70-kD mitochondrial heat shock protein (mt-hsp70) has a dual role in membrane translocation of preproteins we screened preproteins in an attempt to find examples which required either only the unfoldase or only the translocase function of mt-hsp70. We found that a series of fusion proteins containing amino-terminal portions of the intermembrane space protein cytochrome b2 (cyt. b2) fused to dihydrofolate reductase (DHFR) were differentially imported into mitochondria containing mutant hsp70s. A fusion protein between the amino terminal 167 residues of the precursor of cyt. b2 and DHFR was efficiently transported into mitochondria independently of both hsp70 functions. When the length of the cyt. b2 portion was increased and included the heme binding domain, the fusion protein became dependent on the unfoldase function of mt-hsp70, presumably caused by a conformational restriction of the heme-bound preprotein. In the absence of heme the noncovalent heme binding domain in the longer fusion proteins no longer conferred a dependence on the unfoldase function. When the cyt. b2 portion of the fusion protein was less than 167 residues, its import was still independent of mt-hsp70 function; however, deletion of the intermembrane space sorting signal resulted in preproteins that ended up in the matrix of wild type mitochondria and whose translocation was strictly dependent on the translocase function of mt-hsp70. These findings provide strong evidence for a dual role of mt-hsp70 in membrane translocation and indicate that preproteins with an intermembrane space sorting signal can be correctly imported even in mutants with severely impaired hsp70 function. PMID- 8408193 TI - Exofacial epitope-tagged glucose transporter chimeras reveal COOH-terminal sequences governing cellular localization. AB - The insulin-regulated adipocyte/skeletal muscle glucose transporter (GLUT4) displays a characteristic steady-state intracellular localization under basal conditions, whereas the erythrocyte/brain transporter isoform (GLUT1) distributes mostly to the cell surface. To identify possible structural elements in these transporter proteins that determine their cellular localization, GLUT1/GLUT4 chimera cDNA constructs that contain the hemagglutinin epitope YPYDVPDYA (HA) in their major exofacial loops were engineered. Binding of monoclonal anti-HA antibody to non-permeabilized COS-7 cells expressing HA-tagged transporter chimeras revealed that expression of transporters on the cell surface was strongly influenced by their cytoplasmic COOH-terminal domain. This method also revealed a less marked, but significant effect on cellular localization of amino acid residues between transporter exofacial and middle loops. The subcellular distribution of expressed chimeras was confirmed by immunofluorescence microscopy of permeabilized COS-7 cells. Thus, HA-tagged native GLUT4 was concentrated in the perinuclear region, whereas a chimera containing the COOH-terminal 29 residues of GLUT1 substituted onto GLUT4 distributed to the plasma membrane, as did native GLUT1. Furthermore, a chimera composed of GLUT1 with a GLUT4 COOH terminal 30-residue substitution exhibited a predominantly intracellular localization. Similar data was obtained in CHO cells stably expressing these chimeras. Taken together, these results define the unique COOH-terminal cytoplasmic sequences of the GLUT1 and GLUT4 glucose transporters as important determinants of cellular localization in COS-7 and CHO cells. PMID- 8408194 TI - Distinguishing roles of the membrane-cytoskeleton and cadherin mediated cell-cell adhesion in generating different Na+,K(+)-ATPase distributions in polarized epithelia. AB - In simple epithelia, the distribution of ion transporting proteins between the apical or basal-lateral domains of the plasma membrane is important for determining directions of vectorial ion transport across the epithelium. In the choroid plexus, Na+,K(+)-ATPase is localized to the apical plasma membrane domain where it regulates sodium secretion and production of cerebrospinal fluid; in contrast, Na+,K(+)-ATPase is localized to the basal-lateral membrane of cells in the kidney nephron where it regulates ion and solute reabsorption. The mechanisms involved in restricting Na+,K(+)-ATPase distribution to different membrane domains in these simple epithelia are poorly understood. Previous studies have indicated a role for E-cadherin mediated cell-cell adhesion and membrane cytoskeleton (ankyrin and fodrin) assembly in regulating Na+,K(+)-ATPase distribution in absorptive kidney epithelial cells. Confocal immunofluorescence microscopy reveals that in chicken and rat choroid plexus epithelium, fodrin, and ankyrin colocalize with Na+,K(+)-ATPase at the apical plasma membrane, but fodrin, ankyrin, and adducin also localize at the lateral plasma membrane where Na+,K(+)-ATPase is absent. Biochemical analysis shows that fodrin, ankyrin, and Na+,K(+)-ATPase are relatively resistant to extraction from cells in buffers containing Triton X-100. The fractions of Na+,K(+)-ATPase, fodrin, and ankyrin that are extracted from cells cosediment in sucrose gradients at approximately 10.5 S. Further separation of the 10.5 S peak of proteins by electrophoresis in nondenaturing polyacrylamide gels revealed that fodrin, ankyrin, and Na+,K(+) ATPase comigrate, indicating that these proteins are in a high molecular weight complex similar to that found previously in kidney epithelial cells. In contrast, the anion exchanger (AE2), a marker protein of the basal-lateral plasma membrane in the choroid plexus, did not cosediment in sucrose gradients or comigrate in nondenaturing polyacrylamide gels with the complex of Na+,K(+)-ATPase, ankyrin, and fodrin. Ca(++)-dependent cell adhesion molecules (cadherins) were detected at lateral membranes of the choroid plexus epithelium and colocalized with a distinct fraction of ankyrin, fodrin, and adducin. Cadherins did not colocalize with Na+,K(+)-ATPase and were absent from the apical membrane. The fraction of cadherins that was extracted with buffers containing Triton X-100 cosedimented with ankyrin and fodrin in sucrose gradients and comigrated in nondenaturing gels with ankyrin and fodrin in a high molecular weight complex. Since a previous study showed that E-cadherin is an instructive inducer of Na+,K(+)-ATPase distribution, we examined protein distributions in fibroblasts transfected with B cadherin, a prominent cadherin expressed in the choroid plexus epithelium.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408195 TI - Isoform-specific 3'-untranslated sequences sort alpha-cardiac and beta cytoplasmic actin messenger RNAs to different cytoplasmic compartments. AB - We demonstrate that in differentiating myoblasts, the mRNAs encoding two actin isoforms, beta-cytoplasmic, and alpha-cardiac, can occupy different cytoplasmic compartments within the same cytoplasm. beta-actin mRNA is localized to the leading lamellae and alpha-actin mRNA is associated with a perinuclear compartment. This was revealed by co-hybridizing, in situ, fluorochrome conjugated oligonucleotide probes specific for each isoform. To address the mechanism of isoform-specific mRNA localization, molecular chimeras were constructed by insertion of actin sequences between the Lac Z coding region and SV-40 3'UTR in a reporter plasmid. These constructs were transiently expressed in a mixed culture of embryonic fibroblasts, myoblasts and myotubes, beta galactosidase activity within transfectants was revealed by a brief incubation with its substrate (X-gal). Since the blue-insoluble reaction product co localized with the specific mRNAs expressed from each construct, it was used as a bioassay for mRNA localization. Transfectants were scored as either perinuclear, peripheral or nonlocalized with respect to the distribution of the blue product. The percentage of transfectants within those categories was quantitated as a function of the various constructs. This analysis revealed that for each actin mRNA its 3'UTR is necessary and sufficient to direct reporter transcripts to its appropriate compartment; beta-actin peripheral and alpha-actin perinuclear. In contrast, sequences from the 5'UTR through the coding region of either actin gene did not localize the blue product. Therefore, 3'UTR sequences play a key role in modulating the distribution of actin mRNAs in muscle cells. We propose that the mechanism of mRNA localization facilitates actin isoform sorting in the cytoplasm. PMID- 8408196 TI - Localization and dynamics of nonfilamentous actin in cultured cells. AB - Although the distribution of filamentous actin is well characterized in many cell types, the distribution of nonfilamentous actin remains poorly understood. To determine the relative distribution of filamentous and nonfilamentous actin in cultured NRK cells, we have used a number of labeling agents that differ with respect to their specificities toward the filamentous or nonfilamentous form, including monoclonal and polyclonal anti-actin antibodies, vitamin D-binding protein (DBP), and fluorescent phalloidin. Numerous punctate structures were identified that bind poorly to phalloidin but stain positively with several anti actin antibodies. These bead structures also stain with DBP, suggesting that they are enriched in nonfilamentous actin. Similar punctate structures were observed after the microinjection of fluorescently labeled actin into living cells, allowing us to examine their dynamics in living cells. The actin-containing punctate structures were observed predominantly in the region behind lamellipodia, particularly in spreading cells induced by wounding confluent monolayers. Time-lapse recording of cells injected with fluorescent actin indicated that they form continuously near the leading edge and move centripetally toward the nucleus. Our results suggest that at least part of the unpolymerized actin molecules are localized at discrete sites, possibly as complexes with monomer sequestering proteins. These structures may represent transient storage sites of G-actin within the cell which can be transformed rapidly into actin filaments upon stimulation by specific signals. PMID- 8408197 TI - Assembly of flagellar radial spoke proteins in Chlamydomonas: identification of the axoneme binding domain of radial spoke protein 3. AB - Radial spokes of the eukaryotic flagellum extend from the A tubule of each outer doublet microtubule toward the central pair microtubules. In the paralyzed flagella mutant of Chlamydomonas pf14, a mutation in the gene for one of 17 polypeptides that comprise the radial spokes results in flagella that lack all 17 spoke components. The defective gene product, radial spoke protein 3 (RSP3), is, therefore, pivotal to the assembly of the entire spoke and may attach the spoke to the axoneme. We have synthesized RSP3 in vitro and assayed its binding to axonemes from pf14 cells to determine if RSP3 can attach to spokeless axonemes. In vitro, RSP3 binds to pf14 axonemes, but not to wild-type axonemes or microtubules polymerized from purified chick brain tubulin. The sole axoneme binding domain of RSP3 is located within amino acids 1-85 of the 516 amino acid protein; deletion of these amino acids abolishes binding by RSP3. Fusion of amino acids 1-85 or 42-85 to an unrelated protein confers complete or partial binding activity, respectively, to the fusion protein. Transformation of pf14 cells with mutagenized RSP3 genes indicates that amino acids 18-87 of RSP3 are important to its function, but that the carboxy-terminal 140 amino acids can be deleted with little effect on radial spoke assembly or flagellar motility. PMID- 8408198 TI - The relationship between cell size and cell fate in Volvox carteri. AB - In Volvox carteri development, visibly asymmetric cleavage divisions set apart large embryonic cells that will become asexual reproductive cells (gonidia) from smaller cells that will produce terminally differentiated somatic cells. Three mechanisms have been proposed to explain how asymmetric division leads to cell specification in Volvox: (a) by a direct effect of cell size (or a property derived from it) on cell specification, (b) by segregation of a cytoplasmic factor resembling germ plasm into large cells, and (c) by a combined effect of differences in cytoplasmic quality and cytoplasmic quantity. In this study a variety of V. carteri embryos with genetically and experimentally altered patterns of development were examined in an attempt to distinguish among these hypotheses. No evidence was found for regionally specialized cytoplasm that is essential for gonidial specification. In all cases studied, cells with a diameter > approximately 8 microns at the end of cleavage--no matter where or how these cells had been produced in the embryo--developed as gonidia. Instructive observations in this regard were obtained by three different experimental interventions. (a) When heat shock was used to interrupt cleavage prematurely, so that presumptive somatic cells were left much larger than they normally would be at the end of cleavage, most cells differentiated as gonidia. This result was obtained both with wild-type embryos that had already divided asymmetrically (and should have segregated any cytoplasmic determinants involved in cell specification) and with embryos of a mutant that normally produces only somatic cells. (b) When individual wild-type blastomeres were isolated at the 16-cell stage, both the anterior blastomeres that normally produce two gonidia each and the posterior blastomeres that normally produce no gonidia underwent modified cleavage patterns and each produced an average of one large cell that developed as a gonidium. (c) When large cells were created microsurgically in a region of the embryo that normally makes only somatic cells, these large cells became gonidia. These data argue strongly for a central role of cell size in germ/soma specification in Volvox carteri, but leave open the question of how differences in cell size are actually transduced into differences in gene expression. PMID- 8408199 TI - Tissue polarity genes of Drosophila regulate the subcellular location for prehair initiation in pupal wing cells. AB - The Drosophila wing is decorated with a regular array of distally pointing hairs. In the pupal wing, the hairs are formed from micro-villus like prehairs that contain large bundles of actin filaments. The distal orientation of the actin bundles reveals the proximal-distal polarity within the pupal wing epithelium. We have used F-actin staining to examine early stages of prehair development in both wild-type and mutant pupal wings. We have found a striking correlation between hair polarity and the subcellular location for assembly of the prehair. In a wild type wing, all of the distally pointing hairs are derived from prehairs that are formed at the distal vertex of the hexagonally shaped pupal wing cells. Mutations in six tissue polarity genes result in abnormal hair polarity on the adult wing, and all also alter the subcellular location for prehair initiation. Based on their cellular phenotypes, we can place these six genes into three phenotypic groups. Double mutant analysis indicates that these phenotypic groups correspond to epistasis groups. This suggests that the tissue polarity genes function in or on a pathway that controls hair polarity by regulating the subcellular location for prehair formation. PMID- 8408200 TI - Scatter factor/hepatocyte growth factor and its receptor, the c-met tyrosine kinase, can mediate a signal exchange between mesenchyme and epithelia during mouse development. AB - Scatter factor/hepatocyte growth factor (SF/HGF) has potent motogenic, mitogenic, and morphogenetic activities on epithelial cells in vitro. The cell surface receptor for this factor was recently identified: it is the product of the c-met protooncogene, a receptor-type tyrosine kinase. We report here the novel and distinct expression patterns of SF/HGF and its receptor during mouse development, which was determined by a combination of in situ hybridization and RNase protection experiments. Predominantly, we detect transcripts of c-met in epithelial cells of various developing organs, whereas the ligand is expressed in distinct mesenchymal cells in close vicinity. In addition, transient SF/HGF and c met expression is found at certain sites of muscle formation; transient expression of the c-met gene is also detected in developing motoneurons. SF/HGF and the c-met receptor might thus play multiple developmental roles, most notably, mediate a signal given by mesenchyme and received by epithelial. Mesenchymal signals are known to govern differentiation and morphogenesis of many epithelia, but the molecular nature of the signals has remained poorly understood. Therefore, the known biological activities of SF/HGF in vitro and the embryonal expression pattern reported here indicate that this mesenchymal factor can transmit morphogenetic signals in epithelial development and suggest a molecular mechanism for mesenchymal epithelial interactions. PMID- 8408201 TI - The actin-binding protein comitin (p24) is a component of the Golgi apparatus. AB - Comitin (p24) was first identified in Dictyostelium discoideum as a membrane associated protein which binds in gel overlay assays to G and F actin. To analyze its actin-binding properties we used purified, bacterially expressed comitin and found that it binds to F actin in spin down experiments and increases the viscosity of F actin solutions even under high-salt conditions. Immunofluorescence studies, cell fractionation experiments and EM studies of vesicles precipitated with comitin-specific monoclonal antibodies showed that comitin was present in D. discoideum on: (a) a perinuclear structure with tubular or fibrillary extensions; and (b) on vesicles distributed throughout the cell. In immunofluorescence experiments using comitin antibodies NIH 3T3 fibroblasts showed a similar staining pattern as D. discoideum cells. Using bona fide Golgi markers the perinuclear structure was identified as the Golgi apparatus. The results were supported by an electron microscopic study using cryosections. Based on these data we propose that also in Dictyostelium the stained perinuclear structure is the Golgi apparatus. In vivo the perinuclear structure was found to be attached to the actin and the microtubule network. Alteration of the actin network or depolymerization of the microtubules led to its dispersal into vesicles distributed throughout the cell. These results suggest that the Golgi apparatus in D. discoideum is connected to the actin network by comitin. This protein seems also to be present in mammalian cells. PMID- 8408202 TI - Neurons regulate Schwann cell genes by diffusible molecules. AB - Successful peripheral nerve regeneration and functional recovery require the reestablishment of the neuron-Schwann cell relationship in the regenerating rat sciatic nerve, neurons differentially regulate Schwann cell genes. The message for the low-affinity NGF receptor, p75NGFR, is induced in Schwann cells distal to the injury and is repressed as regenerating axons make contact with these cells. The inverse is true for mRNA of the myelin gene P0; expression decreases distal to injury and increases as new axons contact Schwann cells and a program of myelination is initiated. Using an in vitro co-culture paradigm in which primary neurons and adult Schwann cells are separated by a microporous membrane, we show that axon contact is not an absolute requirement for neuronal regulation of Schwann cell genes. In this system neurons but not other cell types, repress the expression of Schwann cell p75NGFR while inducing the expression of the POU domain transcription factor, suppressed cAMP inducible POU, and myelin P0. These results demonstrate that regenerating axons can direct the Schwann cell genetic program from a distance through diffusible molecules. PMID- 8408203 TI - Rab8, a small GTPase involved in vesicular traffic between the TGN and the basolateral plasma membrane. AB - Small GTP-binding proteins of the rab family have been implicated as regulators of membrane traffic along the biosynthetic and endocytic pathways in eukaryotic cells. We have investigated the localization and function of rab8, closely related to the yeast YPT1/SEC4 gene products. Confocal immunofluorescence microscopy and immunoelectron microscopy on filter-grown MDCK cells demonstrated that, rab8 was localized to the Golgi region, vesicular structures, and to the basolateral plasma membrane. Two-dimensional gel electrophoresis showed that rab8p was highly enriched in immuno-isolated basolateral vesicles carrying vesicular stomatitis virus-glycoprotein (VSV-G) but was absent from vesicles transporting the hemagglutinin protein (HA) of influenza virus to the apical cell surface. Using a cytosol dependent in vitro transport assay in permeabilized MDCK cells we studied the functional role of rab8 in biosynthetic membrane traffic. Transport of VSV-G from the TGN to the basolateral plasma membrane was found to be significantly inhibited by a peptide derived from the hypervariable COOH terminal region of rab8, while transport of the influenza HA from the TGN to the apical surface and ER to Golgi transport were unaffected. We conclude that rab8 plays a role in membrane traffic from the TGN to the basolateral plasma membrane in MDCK cells. PMID- 8408204 TI - Protein transport to the dendritic plasma membrane of cultured neurons is regulated by rab8p. AB - In the companion paper (Huber, L. A., S. W. Pimplikar, R. G. Parton, H. Virta, M. Zerial, and K. Simons. J. Cell Biol. 123:35-45) we reported that the small GTPase rab8p is involved in transport from the TGN to the basolateral plasma membrane in epithelia. In the present work we investigated the localization and function of rab8p in polarized hippocampal neurons. By immunofluorescence microscopy we found that rab8p localized preferentially in the somatodendritic domain, and was excluded from the axon. Double-labeling immunofluorescence showed that some of the rab8p co-localized in the dendrites with the Semliki Forest Virus glycoprotein E2 (SFV-E2). An antisense oligonucleotide approach was used to investigate the role of rab8p in dendritic transport of newly synthesized viral glycoproteins. Antisense oligonucleotides corresponding to the initiation region of the rab8 coding sequence were added to the cultured neurons for four days. This treatment resulted in a significant decrease in cellular levels of rab8p and transport of SFV-E2 from the cell body to the dendrites was significantly reduced. However, no effect was observed on axonal transport of influenza HA. From these results we conclude that rab8p is involved in transport of proteins to the dendritic surface in neurons. PMID- 8408205 TI - Cholesterol is required in the exit pathway of Semliki Forest virus. AB - The enveloped alphavirus Semliki Forest virus (SFV) infects cells via a membrane fusion reaction triggered by low pH. For fusion to occur cholesterol is required in the target membrane, as demonstrated both in in vitro fusion assays and in vivo for virus infection of a host cell. In this paper we examine the role of cholesterol in postfusion events in the SFV life cycle. Cholesterol-depleted insect cells were transfected with SFV RNA or infected at very high multiplicities to circumvent the fusion block caused by the absence of cholesterol. Under these conditions, the viral spike proteins were synthesized and transported to the site of p62 cleavage with normal kinetics. Surprisingly, the subsequent exit of virus particles was dramatically slowed compared to cholesterol-containing cells. The inhibition of virus production could be reversed by the addition of cholesterol to depleted cells. In contrast to results with SFV, no cholesterol requirement for virus exit was observed for the production of either the unrelated vesicular stomatitis virus or a cholesterol independent SFV fusion mutant. Thus, cholesterol was only critical in the exit pathway of viruses that also require cholesterol for fusion. These results demonstrate a specific and unexpected lipid requirement in virus exit, and suggest that in addition to its role in fusion, cholesterol is involved in the assembly or budding of SFV. PMID- 8408206 TI - Assembly and targeting of adaptin chimeras in transfected cells. AB - Adaptors are the components of clathrincoated pits and vesicles that attach the clathrin to the membrane. There are two types of adaptors in the cell: one associated with the plasma membrane and one associated with the TGN. Both adaptors are heterotetramers consisting of two adaptins (alpha and beta for the plasma membrane; gamma and beta' for the TGN), plus two smaller proteins. The COOH-terminal domains of the adaptins form appendages that resemble ears, connected by flexible hinges. Unlike the other adaptor components, the COOH termini of the alpha- and gamma-adaptins show no homology with each other, suggesting that they might provide the signal that directs the adaptors to the appropriate membrane. To test this possibility, the COOH-terminal ears were switched between alpha- and gamma-adaptins and were also deleted. All of the constructs contained the bovine gamma-adaptin hinge, enabling them to be detected with a species-specific antibody against this region when transfected into rat fibroblasts. Immunoprecipitation indicated that the engineered adaptins were still fully capable of assembling into adaptor complexes. Immunofluorescence revealed that in spite of their modified ears, the constructs were still able to be recruited onto the appropriate membrane; however, the ear-minus constructs gave increased cytoplasmic staining, and replacing the gamma-adaptin ear with the alpha-adaptin ear caused a small amount of colocalization with endogenous alpha adaptin in some cells. Thus, the major targeting determinant appears to reside in the adaptor "head," while the ears may stabilize the association of adaptors with the membrane. PMID- 8408207 TI - Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis. AB - We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated nuclei. The system uses extracts prepared from mitotic chicken hepatoma cells following a sequential S phase/M phase synchronization. When nuclei are added to these extracts, the chromatin becomes highly condensed into spherical domains that ultimately extrude through the nuclear envelope, forming apoptotic bodies. The process is highly synchronous, and the structural changes are completed within 60 min. Coincident with these morphological changes, the nuclear DNA is cleaved into a nucleosomal ladder. Both processes are inhibited by Zn2+, an inhibitor of apoptosis in intact cells. Nuclear lamina disassembly accompanies these structural changes in added nuclei, and we show that lamina disassembly is a characteristic feature of apoptosis in intact cells of mouse, human and chicken. This system may provide a powerful means of dissecting the biochemical mechanisms underlying the final stages of apoptosis. PMID- 8408208 TI - Redistribution of clathrin-coated vesicle adaptor complexes during adipocytic differentiation of 3T3-L1 cells. AB - Mechanisms for intracellular retention of proteins are induced during adipocytic differentiation of 3T3-L1 cells. To investigate the potential role of clathrin lattices in these retention processes, we performed a morphological and biochemical analysis of coated vesicle components in 3T3-L1 cells. Optical sectioning and image restoration revealed a marked increase in the staining of clathrin and beta adaptins in the perinuclear region of cells with differentiation. In addition, predominance of beta (subunit of the AP-2, plasma membrane adaptor) over beta' (subunit of the AP-1, Golgi adaptor) adaptin was observed in immunoblots of clathrin-coated vesicles purified from nondifferentiated fibroblasts, and this ratio was reversed in coated vesicles purified from differentiated adipocytes. These results indicate that the relative abundance of TGN-derived clathrin lattices increases markedly during adipocytic differentiation. Subcellular fractionation indicated that cytosolic AP-1 and AP-2 adaptors comprised approximately 70% of the total cellular adaptor pool. Interestingly, neither the concentration nor the relative ratio of cytosolic AP-1 to AP-2 adaptors increased significantly during differentiation. These data suggest that the increase in TGN-derived lattices results from differentiation induced mechanisms for enhanced assembly or stabilization of adaptors on Golgi membranes. Interestingly, double-immunofluorescence microscopy also revealed that whereas extensive colocalization between clathrin and beta adaptins occurred both in fibroblasts and adipocytes, structures stained only with anti-adaptin antibody could be detected. Taken together these results suggest that membranes coated with adaptors, but not clathrin, can exist in these cells. PMID- 8408209 TI - Molecules internalized by clathrin-independent endocytosis are delivered to endosomes containing transferrin receptors. AB - We have previously demonstrated that the preendosomal compartment in addition to clathrin-coated vesicles, comprises distinct nonclathrin coated endocytic vesicles mediating clathrin-independent endocytosis (Hansen, S. H., K. Sandvig, and B. van Deurs. 1991. J. Cell Biol. 113:731-741). Using K+ depletion in HEp-2 cells to block clathrin-dependent but not clathrin-independent endocytosis, we have now traced the intracellular routing of these nonclathrin coated vesicles to see whether molecules internalized by clathrin-independent endocytosis are delivered to a unique compartment or whether they reach the same early and late endosomes as encountered by molecules internalized with high efficiency through clathrin-coated pits and vesicles. We find that Con A-gold internalized by clathrin-independent endocytosis is delivered to endosomes containing transferrin receptors. After incubation of K(+)-depleted cells with Con A-gold for 15 min, approximately 75% of Con A-gold in endosomes is colocalized with transferrin receptors. Endosomes containing only Con A-gold may be accounted for either by depletion of existing endosomes for transferrin receptors or by de novo generation of endosomes. Cationized gold and BSA-gold internalized in K(+) depleted cells are also delivered to endosomes containing transferrin receptors. h-lamp-1-enriched compartments are only reached occasionally within 30 min in K(+)-depleted as well as in control cells. Thus, preendosomal vesicles generated by clathrin-independent endocytosis do not fuse to any marked degree with late endocytic compartments. These data show that in HEp-2 cells, molecules endocytosed without clathrin are delivered to the same endosomes as reached by transferrin receptors internalized through clathrin-coated pits. PMID- 8408210 TI - Mannose 6-phosphate-independent targeting of lysosomal enzymes in I-cell disease B lymphoblasts. AB - B lymphocytes from patients with I-cell disease (ICD) maintain normal cellular levels of lysosomal enzymes despite a deficiency of the enzyme UDP-N acetylglucosamine: lysosomal enzyme N-acetylglucosamine-1-phosphotransferase. We find that an ICD B lymphoblastoid cell line targets about 45% of the lysosomal protease cathepsin D to dense lysosomes. This targeting occurs in the absence of detectable mannose 6-phosphate residues on the cathepsin D and is not observed in ICD fibroblasts. The secretory protein pepsinogen, which is closely related to cathepsin D in both amino acid sequence and three-dimensional structure, is mostly excluded from dense lysosomes, indicating that the lymphoblast targeting pathway is specific. Carbohydrate residues are not required for lysosomal targeting, since a non-glycosylated mutant cathepsin D is sorted with comparable efficiency to the wild type protein. Analysis of a number of cathepsin D/pepsinogen chimeric proteins indicates that an extensive polypeptide determinant in the cathepsin D carboxyl lobe can confer efficient lysosomal sorting when introduced into the pepsinogen sequence. This determinant overlaps but is not identical to the recognition marker for phosphotransferase. These results indicate that a specific protein recognition event underlies Man-6-P independent lysosomal sorting in ICD lymphoblasts. PMID- 8408211 TI - RNA on the move: the mRNA localization pathway. PMID- 8408212 TI - Proteolytic cleavage of haptoglobin occurs in a subcompartment of the endoplasmic reticulum: evidence from membrane fusion in vitro. AB - The primary translation product of haptoglobin mRNA is a 45-kD polypeptide which is proteolytically cleaved shortly after its synthesis. Previous studies have indicated that the cleavage of this proform of haptoglobin occurs in the ER. In an attempt to characterize the cleaving enzyme, we found that upon incubation of microsomes from rat hepatocytes pulse labeled with [35S]methionine, little cleavage of labeled prohaptoglobin occurred. In contrast, when cells whose cytoplasmic proteins had been released by saponin treatment were incubated, 30 40% of the prohaptoglobin was cleaved. The addition of GTP caused a twofold stimulation, which was abolished by the nonhydrolyzable analog GTP gamma S. With a homogenate of the cells, the addition of GTP resulted in a fourfold stimulation of the degree of cleavage--from 15 to 60%. Differential centrifugation revealed that most of the cleaving activity resided in membranes sedimenting similarly to mitochondria and to a small fraction of the ER. These rapidly sedimenting membranes were therefore prepared from a rat liver homogenate. Upon treatment with high salt, light membranes were released which, when incubated with microsomes of pulse-labeled hepatocytes in the presence of detergent (and in the absence of GTP), induced specific cleavage of prohaptoglobin. The cleaving enzyme had an alkaline pH optimum indicating that it was not of lysosomal origin. These results suggest that cleavage of prohaptoglobin occurs in a subcompartment of the ER. Apparently, the connection between this compartment and the bulk of the ER is broken upon saponin treatment or homogenization but can be reestablished through a process requiring GTP hydrolysis. PMID- 8408213 TI - Assembly of the tight junction: the role of diacylglycerol. AB - Extracellular Ca2+ triggers assembly and sealing of tight junctions (TJs) in MDCK cells. These events are modulated by G-proteins, phospholipase C, protein kinase C (PKC), and calmodulin. In the present work we observed that 1,2 dioctanoylglycerol (diC8) promotes the assembly of TJ in low extracellular Ca2+, as evidenced by translocation of the TJ-associated protein ZO-1 to the plasma membrane, formation of junctional fibrils observed in freeze-fracture replicas, decreased permeability of the intercellular space to [3H]mannitol, and reorganization of actin filaments to the cell periphery, visualized by fluorescence microscopy using rhodamine-phalloidin. In contrast, diC8 in low Ca2+ did not induce redistribution of the Ca-dependent adhesion protein E-cadherin (uvomorulin). Extracellular antibodies to E-cadherin block junction formation normally induced by adding Ca2+. diC8 counteracted this inhibition, suggesting that PKC may be in the signaling pathway activated by E-cadherin-mediated cell cell adhesion. In addition, we found a novel phosphoprotein of 130 kD which coimmunoprecipitated with the ZO-1/ZO-2 complex. Although the assembly and sealing of TJs may involve the activation of PKC, the level of phosphorylation of ZO-1, ZO-2, and the 130-kD protein did not change after adding Ca2+ or a PKC agonist. The complex of these three proteins was present even in low extracellular Ca2+, suggesting that the addition of Ca2+ or diC8 triggers the translocation and assembly of preformed TJ subcomplexes. PMID- 8408214 TI - Thick filament substructures in Caenorhabditis elegans: evidence for two populations of paramyosin. AB - The thick filaments of the nematode Caenorhabditis elegans contain two myosin heavy chain isoforms A and B and paramyosin, the products of the myo-3, unc-54, and unc-15 genes, respectively. Dissociation of paramyosin from native thick filaments at pH 6.36 shows a biphasic function with respect to NaCl concentration. Electron microscopy of the remaining structures shows 15-nm core structures that label with monoclonal anti-paramyosin antibody at 72.5-nm intervals. Purified core structures also show 72.5 nm repeats by negative staining. Structural analysis of native thick filaments and dissociated structures suggests that the more dissociable paramyosin is removed radially as well as processively from the filament ends. Minor proteins with masses of 20, 28, and 30 kD cosediment stoichiometrically with paramyosin in purified core structures. PMID- 8408215 TI - Three-dimensional reconstruction of caldesmon-containing smooth muscle thin filaments. AB - Caldesmon is known to inhibit actomyosin ATPase and filament sliding in vitro, and may play a role in modulating smooth muscle contraction as well as in diverse cellular processes including cytokinesis and exocytosis. However, the structural basis of caldesmon action has not previously been apparent. We have recorded electron microscope images of negatively stained thin filaments containing caldesmon and tropomyosin which were isolated from chicken gizzard smooth muscle in EGTA. Three-dimensional helical reconstructions of these filaments show actin monomers whose bilobed shape and connectivity are very similar to those previously seen in reconstructions of frozen-hydrated skeletal muscle thin filaments. In addition, a continuous thin strand of density follows the long pitch actin helices, in contact with the inner domain of each actin monomer. Gizzard thin filaments treated with Ca2+/calmodulin, which dissociated caldesmon but not tropomyosin, have also been reconstructed. Under these conditions, reconstructions also reveal a bilobed actin monomer, as well as a continuous surface strand that appears to have moved to a position closer to the outer domain of actin. The strands seen in both EGTA- and Ca2+/calmodulin-treated filaments thus presumably represent tropomyosin. It appears that caldesmon can fix tropomyosin in a particular position on actin in the absence of calcium. An influence of caldesmon on tropomyosin position might, in principle, account for caldesmon's ability to modulate actomyosin interaction in both smooth muscles and non-muscle cells. PMID- 8408216 TI - Contractile activity and cell-cell contact regulate myofibrillar organization in cultured cardiac myocytes. AB - Adult feline ventricular myocytes cultured on a laminin-coated substratum reestablish intercellular junctions, yet disassemble their myofibrils. Immunofluorescence microscopy reveals that these non-beating heart cells lack vinculin-positive focal adhesions; moreover, intercellular junctions are also devoid of vinculin. When these quiescent myocytes are stimulated to contract with the beta-adrenergic agonist, isoproterenol, extensive vinculin-positive focal adhesions and intercellular junctions emerge. If solitary myocytes are stimulated to beat, an elaborate series of vinculin-positive focal adhesions develop which appear to parallel the reassembly of myofibrils. In cultures where neighboring myocytes reestablish cell-cell contact, myofibrils appear to reassemble from the fascia adherens rather than focal contacts. Activation of beating is accompanied by a significant reduction in the rate of total and cytoskeletal protein synthesis; in fact, myofibrillar reassembly, redevelopment of focal adhesions and fascia adherens junctions require no protein synthesis for at least 24 h, implying the existence of an assembly competent pool of cytoskeletal proteins. Maturation of the fasciae adherens and the appearance of vinculin within Z line/costameres, does require de novo synthesis of new cytoskeletal proteins. Changes in cytoskeletal protein turnover appear dependent on beta agonist-induced cAMP production, but myofibrillar reassembly is a cAMP-independent event. Such observations suggest that mechanical forces, in the guise of contractile activity, regulate vinculin distribution and myofibrillar order in cultured adult feline heart cells. PMID- 8408217 TI - A 13-A map of the actin-scruin filament from the limulus acrosomal process. AB - We have determined the structure of the actin-scruin filament to 13-A resolution using a combination of low-dose EM and image analysis. The three-dimensional map reveals four actin-actin contacts: two within each strand and two between strands. The conformation of the actin subunit is different from that in the Holmes et al. (1990) model as refined by Lorenz et al. (1993). In particular, subdomain II is tilted in a similar way to that seen by Orlova and Egelman (1993) in F-Mg2(+)-ADP actin filaments in the absence of Ca2+. Scruin appears to consist of two domains of approximately equal volume. Each scruin subunit cross-links the two strands in the actin filament. Domain I of scruin contacts subdomain I of actin and makes a second contact at the junction of subdomains III and IV. Domain II of scruin contacts actin at subdomains I and II of a neighboring actin subunit. The two scruin domains thus bind differently to actin. PMID- 8408218 TI - In vitro models of tail contraction and cytoplasmic streaming in amoeboid cells. AB - We have developed a reconstituted gel-sol and contractile model system that mimics the structure and dynamics found at the ectoplasm/endoplasm interface in the tails of many amoeboid cells. We tested the role of gel-sol transformations of the actin-based cytoskeleton in the regulation of contraction and in the generation of endoplasm from ectoplasm. In a model system with fully phosphorylated myosin II, we demonstrated that either decreasing the actin filament length distribution or decreasing the extent of actin filament cross linking initiated both a weakening of the gel strength and contraction. However, streaming of the solated gel components occurred only under conditions where the length distribution of actin was decreased, causing a self-destruct process of continued solation and contraction of the gel. These results offer significant support that gel strength plays an important role in the regulation of actin/myosin II-based contractions of the tail cortex in many amoeboid cells as defined by the solation-contraction coupling hypothesis (Taylor, D. L., and M. Fechheimer. 1982. Phil. Trans. Soc. Lond. B. 299:185-197). The competing processes of solation and contraction of the gel would appear to be mutually exclusive. However, it is the temporal-spatial balance of the rate and extent of two stages of solation, coupled to contraction, that can explain the conversion of gelled ectoplasm in the tail to a solated endoplasm within the same small volume, generation of a force for the retraction of tails, maintenance of cell polarity, and creation of a positive hydrostatic pressure to push against the newly formed endoplasm. The mechanism of solation-contraction of cortical cytoplasm may be a general component of the normal movement of a variety of amoeboid cells and may also be a component of other contractile events such as cytokinesis. PMID- 8408219 TI - Identification and molecular characterization of E-MAP-115, a novel microtubule associated protein predominantly expressed in epithelial cells. AB - A novel microtubule-associated protein (MAP) of M(r) 115,000 has been identified by screening of a HeLa cell cDNA expression library with an anti-serum raised against microtubule-binding proteins from HeLa cells. Monoclonal and affinity purified polyclonal antibodies were generated for the further characterization of this MAP. It is different from the microtubule-binding proteins of similar molecular weights, characterized so far, by its nucleotide-insensitive binding to microtubules and different sedimentation behavior. Since it is predominantly expressed in cells of epithelial origin (Caco-2, HeLa, MDCK), and rare (human skin, A72) or not detectable (Vero) in fibroblastic cells, we name it E-MAP-115 (epithelial MAP of 115 kD). In HeLa cells, E-MAP-115 is preferentially associated with subdomains or subsets of perinuclear microtubules. In Caco-2 cells, labeling for E-MAP-115 increases when they polarize and form blisters. The molecular characterization of E-MAP-115 reveals that it is a novel protein with no significant homologies to other known proteins. The secondary structure predicted from its sequence indicates two domains connected by a putative hinge region rich in proline and alanine (PAPA region). E-MAP-115 has two highly charged regions with predicted alpha-helical structure, one basic with a pI of 10.9 in the NH2 terminal domain and one neutral with a pI of 7.6 immediately following the PAPA region in the acidic COOH-terminal half of the molecule. A novel microtubule binding site has been localized to the basic alpha-helical region in the NH2 terminal domain using in vitro microtubule-binding assays and expression of mutant polypeptides in vivo. Overexpression of this domain of E-MAP-115 by transfection of fibroblasts lacking significant levels of this protein with its cDNA renders microtubules stable to nocodazole. We conclude that E-MAP-115 is a microtubule-stabilizing protein that may play an important role during reorganization of microtubules during polarization and differentiation of epithelial cells. PMID- 8408220 TI - Molecular analysis of the INCENPs (inner centromere proteins): separate domains are required for association with microtubules during interphase and with the central spindle during anaphase. AB - It has recently been proposed that mitotic chromosomes transport certain cytoskeletal proteins to the metaphase plate so that these proteins are able to subsequently participate in the assembly of the anaphase spindle and the cleavage furrow. To understand how such proteins accomplish their dual chromosomal: cytoskeletal role, we have begun a molecular and functional analysis of the inner centromere proteins (INCENPs), founder members of the class of "chromosome passenger proteins". cDNA clones encoding the open reading frames of the two chicken INCENPs were recovered. The predicted proteins, class I INCENP (96,357 D) and class II INCENP (100,931 D) are novel, and differ from each other by the inclusion of a 38-codon insert within the class II coding region. Transient expression of the chicken INCENPs in mammalian cells confirms that the signals and structures required for the transfer of these proteins from chromosomes to cytoskeleton are evolutionarily conserved. Furthermore, these studies reveal that INCENP association with the cytoskeleton is complex. The amino-terminal 42-amino acid residues are required for transfer of the INCENPs from the chromosomes to the mitotic spindle at anaphase, but not for binding of INCENPs to cytoplasmic microtubules. In contrast, an internal 200 amino acid coiled-coil domain was required for association with microtubules, but dispensable for spindle association. These experiments suggest that proteins required for assembly of specialized cytoskeletal structures during mitosis from anaphase onwards might be sequestered in the nucleus throughout interphase to keep them from disrupting the interphase cytoskeleton, and to ensure their correct positioning during mitosis. PMID- 8408221 TI - MIF2 is required for mitotic spindle integrity during anaphase spindle elongation in Saccharomyces cerevisiae. AB - The function of the essential MIF2 gene in the Saccharomyces cerevisiae cell cycle was examined by overepressing or creating a deficit of MIF2 gene product. When MIF2 was overexpressed, chromosomes missegregated during mitosis and cells accumulated in the G2 and M phases of the cell cycle. Temperature sensitive mutants isolated by in vitro mutagenesis delayed cell cycle progression when grown at the restrictive temperature, accumulated as large budded cells that had completed DNA replication but not chromosome segregation, and lost viability as they passed through mitosis. Mutant cells also showed increased levels of mitotic chromosome loss, supersensitivity to the microtubule destabilizing drug MBC, and morphologically aberrant spindles. mif2 mutant spindles arrested development immediately before anaphase spindle elongation, and then frequently broke apart into two disconnected short half spindles with misoriented spindle pole bodies. These findings indicate that MIF2 is required for structural integrity of the spindle during anaphase spindle elongation. The deduced Mif2 protein sequence shared no extensive homologies with previously identified proteins but did contain a short region of homology to a motif involved in binding AT rich DNA by the Drosophila D1 and mammalian HMGI chromosomal proteins. PMID- 8408222 TI - The calcium-binding protein cell division cycle 31 of Saccharomyces cerevisiae is a component of the half bridge of the spindle pole body. AB - Cdc31 mutants of Saccharomyces cerevisiae arrest at the nonpermissive temperature with large buds, G2 DNA content and, a single, abnormally large spindle pole body (SPB) (Byers, B. 1981. Molecular Genetics in Yeast. Alfred Benzon Symposium. 16:119-133). In this report, we show that the CDC31 gene product is essential for cell viability. We demonstrate that purified CDC31 protein binds Ca2+ and that this binding is highly specific. Taken together, three lines of evidence indicate that CDC31 is a component of the SPB. First, CDC31 cofractionates with enriched preparations of SPBs. Second, immunofluorescence staining indicates that CDC31 colocalizes with a known SPB component. Third, immunoelectron microscopy with whole cells and with isolated SPBs reveals that CDC31 is localized to the half bridge of the SPB, which lies immediately adjacent to the SPB plaques. CDC31 was detected mainly at the cytoplasmic side of the half bridge and, therefore, defines a further substructure of the SPB. We suggest that CDC31 is a member of a family of calcium-binding, centrosome-associated proteins from a phylogenetically diverse group of organisms. PMID- 8408223 TI - Depletion of 43-kD growth-associated protein in primary sensory neurons leads to diminished formation and spreading of growth cones. AB - The 43-kD growth-associated protein (GAP-43) is a major protein kinase C (PKC) substrate of growing axons, and of developing nerve terminals and glial cells. It is a highly hydrophilic protein associated with the cortical cytoskeleton and membranes. In neurons it is rapidly transported from the cell body to growth cones and nerve terminals, where it accumulates. To define the role of GAP-43 in neurite outgrowth, we analyzed neurite regeneration in cultured dorsal root ganglia (DRG) neurons that had been depleted of GAP-43 with any of three nonoverlapping antisense oligonucleotides. The GAP-43 depletion procedure was specific for this protein and an antisense oligonucleotide to the related PKC substrate MARCKS did not detectably affect GAP-43 immunoreactivity. We report that neurite outgrowth and morphology depended on the levels of GAP-43 in the neurons in a substrate-specific manner. When grown on a laminin substratum, GAP 43-depleted neurons extended longer, thinner and less branched neurites with strikingly smaller growth cones than their GAP-43-expressing counterparts. In contrast, suppression of GAP-43 expression prevented growth cone and neurite formation when DRG neurons were plated on poly-L-ornithine. These findings indicate that GAP-43 plays an important role in growth cone formation and neurite outgrowth. It may be involved in the potentiation of growth cone responses to external signals affecting process formation and guidance. PMID- 8408224 TI - A myelin proteolipid protein-LacZ fusion protein is developmentally regulated and targeted to the myelin membrane in transgenic mice. AB - Transgenic mice were generated with a fusion gene carrying a portion of the murine myelin proteolipid protein (PLP) gene, including the first intron, fused to the E. coli LacZ gene. Three transgenic lines were derived and all lines expressed the transgene in central nervous system white matter as measured by a histochemical assay for the detection of beta-galactosidase activity. PLP-LacZ transgene expression was regulated in both a spatial and temporal manner, consistent with endogenous PLP expression. Moreover, the transgene was expressed specifically in oligodendrocytes from primary mixed glial cultures prepared from transgenic mouse brains and appeared to be developmentally regulated in vitro as well. Transgene expression occurred in embryos, presumably in pre- or nonmyelinating cells, rather extensively throughout the peripheral nervous system and within very discrete regions of the central nervous system. Surprisingly, beta-galactosidase activity was localized predominantly in the myelin in these transgenic animals, suggesting that the NH2-terminal 13 amino acids of PLP, which were present in the PLP-LacZ gene product, were sufficient to target the protein to the myelin membrane. Thus, the first half of the PLP gene contains sequences sufficient to direct both spatial and temporal gene regulation and to encode amino acids important in targeting the protein to the myelin membrane. PMID- 8408225 TI - Differential expression of mRNAs for neurotrophins and their receptors after axotomy of the sciatic nerve. AB - The neurotrophin family includes NGF, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and neurotrophin-4 (NT-4). Previous studies have demonstrated that expression of NGF and its low-affinity receptor is induced in nonneuronal cells of the distal segment of the transected sciatic nerve suggesting a role for NGF during axonal regeneration (Johnson, E. M., M. Taniuchi, and P. S. DeStefano. 1988. Trends Neurosci. 11:299-304). To assess the role of the other neurotrophins and the members of the family of Trk signaling neurotrophin receptors, we have here quantified the levels of mRNAs for BDNF, NT 3, and NT-4 as well as mRNAs for trkA, trkB, and trkC at different times after transection of the sciatic nerve in adult rats. A marked increase of BDNF and NT 4 mRNAs in the distal segment of the sciatic nerve was seen 2 wk after the lesion. The increase in BDNF mRNA was mediated by a selective activation of the BDNF exon IV promoter and adrenalectomy attenuated this increase by 50%. NT-3 mRNA, on the other hand, decreased shortly after the transection but returned to control levels 2 wk later. In Schwann cells ensheathing the sciatic nerve, only trkB mRNA encoding truncated TrkB receptors was detected with reduced levels in the distal part of the lesioned nerve. Similar results were seen using a probe that detects all forms of trkC mRNA. In the denervated gastrocnemius muscle, the level of BDNF mRNA increased, NT-3 mRNA did not change, while NT-4 mRNA decreased. In the spinal cord, only small changes were seen in the levels of neutrophin and trk mRNAs. These results show that expression of mRNAs for neurotrophins and their Trk receptors is differentially regulated after a peripheral nerve injury. Based on these results a model is presented for how the different neurotrophins could cooperate to promote regeneration of injured peripheral nerves. PMID- 8408226 TI - Human carcinoembryonic antigen, an intercellular adhesion molecule, blocks fusion and differentiation of rat myoblasts. AB - Human carcinoembryonic antigen (CEA), a widely used tumor marker, is a member of a family of cell surface glycoproteins that are overexpressed in many carcinomas. CEA has been shown to function in vitro as a homotypic intercellular adhesion molecule. This correlation of overproduction of an adhesion molecule with neoplastic transformation provoked a test of the effect of CEA on cell differentiation. Using stable CEA transfectants of the rat L6 myoblast cell line as a model system of differentiation, we show that fusion into myotubes and, in fact, the entire molecular program of differentiation, including creatine phosphokinase upregulation, myogenin upregulation, and beta-actin downregulation are completely abrogated by the ectopic expression of CEA. The blocking of the upregulation of myogenin, a transcriptional regulator responsible for the execution of the entire myogenic differentiation program, indicates that CEA expression intercepts the process at a very early stage. The adhesion function of CEA is essential for this effect since an adhesion-defective N domain deletion mutant of CEA was ineffective in blocking fusion and CEA transfectants treated with adhesion-blocking peptides fused normally. Furthermore, CEA transfectants maintain their high division potential, whereas control transfectants lose division potential with differentiation similarly to the parental cell line. Thus the expression of functional CEA on the surface of cells can block terminal differentiation and maintain proliferative potential. PMID- 8408227 TI - Induction of a secondary body axis in Xenopus by antibodies to beta-catenin. AB - We have obtained evidence that a known intracellular component of the cadherin cell-cell adhesion machinery, beta-catenin, contributes to the development of the body axis in the frog Xenopus laevis. Vertebrate beta-catenin is homologous to the Drosophila segment polarity gene product armadillo, and to vertebrate plakoglobin (McCrea, P. D., C. W. Turck, and B. Gumbiner. 1991. Science (Wash. DC). 254: 1359-1361.). Beta-Catenin was found present in all Xenopus embryonic stages examined, and associated with C-cadherin, the major cadherin present in early Xenopus embryos. To test beta-catenin's function, affinity purified Fab fragments were injected into ventral blastomeres of developing four-cell Xenopus embryos. A dramatic phenotype, the duplication of the dorsoanterior embryonic axis, was observed. Furthermore, Fab injections were capable of rescuing dorsal features in UV-ventralized embryos. Similar phenotypes have been observed in misexpression studies of the Wnt and other gene products, suggesting that beta catenin participates in a signaling pathway which specifies embryonic patterning. PMID- 8408228 TI - Bradykinin-stimulated transient modulation of epidermal growth factor receptors in A-431 human epidermoid carcinoma cells. AB - Of nine biological factors (ATP, bradykinin, vasopressin, substance P, angiotensin II, norepinephrine, epinephrine, 12-tetradecanoylphorbol 13-acetate (TPA), and A23187 calcium ionophore) examined, bradykinin, as well as ATP, TPA, and A23187, significantly increased the phosphorylation of epidermal growth factor (EGF) receptors and reduced the binding of EGF to their high-affinity site. The reduction in EGF binding by bradykinin, ATP, and TPA was similarly reversed by concomitant incubation with staurosporine, a protein kinase C inhibitor, implying that the phosphorylation of EGF receptors was catalyzed probably by a protein kinase C of the same or similar type in each case. This possibility was confirmed by the fact that the major phosphorylation site of EGF receptors by the stimulation with either bradykinin, ATP, or TPA was the same (Thr-654). Different from the stimulations with ATP and TPA, the effect of bradykinin of decreasing the high-affinity EGF binding was transient (a minimum binding at 2.5 min); the reduced EGF binding was, however, sustained for up to 30 min in the presence of calyculin A, a phosphoprotein phosphatase inhibitor. Moreover, the homogenate prepared from bradykinin-stimulated A-431 cells had stronger dephosphorylation activity for phosphorylated EGF receptors than that from control cells. These results suggest that bradykinin stimulates both the protein kinase C system and a phosphoprotein phosphatase(s) activity in A-431 cells. Such biphasic effects of bradykinin to phosphorylate and dephosphorylate EGF receptors via protein kinase C and a phosphoprotein phosphatase, respectively, imply a homeostatic control of receptor function in regulating phosphorylation level by the same bioactive factor. PMID- 8408229 TI - In vitro DNA synthesis of mouse hepatocytes stimulated by tumor necrosis factor is inhibited by glucocorticoids and prostaglandin D2 but enhanced by retinoic acid. AB - We have recently shown that TNF is produced in liver rapidly after partial hepatectomy and that TNF can stimulate DNA synthesis of hepatocyte primary culture with its inhibition by interleukin 6 and transforming growth factor-beta, indicating a pivotal role of TNF and TNF-driven cytokine induction in liver regeneration. We here examined the effects of biological or inflammatory mediators of low molecular weight on the in vitro DNA synthesis of hepatocytes stimulated by TNF. Simultaneous addition of dexamethasone markedly suppressed the growth-stimulating action of TNF, maximally at 10(-7) M and effectively at about 10(-8) M. However, the growth-stimulating effect of EGF was not affected by dexamethasone at all. Physiological glucocorticoids, corticosterone, and hydrocortisone showed virtually the same effect, but other steroid hormones, beta estradiol, or progesterone did not. Retinoic acid at 10(-7) M, however, enhanced TNF-stimulated hepatocyte DNA synthesis and even more effectively the growth response to EGF. PGD2 at 20 microM was markedly suppressive but PGE2 was not. The addition of indomethacin enhanced the hepatocyte in vitro growth by TNF and EGF at 2-20 microM. These results indicate that the growth of hepatocytes stimulated by TNF is up- and down-regulated by inflammatory mediators and hormones as well as cytokines and suggest the biological significance of TNF and TNF-driven inflammatory reactions in liver regeneration. PMID- 8408230 TI - Increased epidermal growth factor receptor in an estrogen-responsive, adriamycin resistant MCF-7 cell line. AB - We examined the expression of the estrogen and epidermal growth factor (EGF) receptors in a drug-resistant subline of MCF-7 cells in order to study potential alterations in hormone dependence or in the growth factor pathway that could be related to the development of drug resistance in human breast cancer. The drug resistant subline was derived from MCF-7 cells by selection with Adriamycin in the presence of the P-glycoprotein antagonist, verapamil, to prevent acquisition of the classical multidrug resistance phenotype. The Adriamycin-resistant cells retain estrogen-binding, estrogen-responsive monolayer growth, and estrogen dependent tumorigenesis. Estrogen-binding studies demonstrate 1.4 x 10(6) sites per cell with unaltered affinity when compared to parental MCF-7 cells, which have 2.7 x 10(5) sites per cell. An increase in expression of EGF receptor, eight to 12-fold, occurred early in the selection for drug resistance, and appears to be unrelated to verapamil exposure, since cells maintained in Adriamycin without verapamil also have increased EGF receptor expression. Partially drug-sensitive revertants carried a verapamil, but out of Adriamycin, demonstrate a decline in EGF receptor expression. We postulate that activation of growth factor pathways in drug-resistant cells may enhance mechanisms of drug resistance, or provide mitogenic stimuli for cells to recover after damage by drug exposure. PMID- 8408231 TI - Effects of ultraviolet irradiation on human skin-derived epidermal cells in vitro. AB - The effects of UVA, mixed UVA + B, and solar-simulated irradiation were examined in human keratinocytes and melanocytes cultured in vitro. Irradiation with UVA, UVA + B, or the solar simulator caused a dose-dependent decrease in keratinocyte cell numbers and thymidine incorporation at 24 hours, with recovery after 48 and 72 hours. Divided dose regimens reduced the inhibitory effect of ultraviolet (UV) irradiation on cell numbers measured 24 hours after the last irradiation. Exposure to both UVA and UVA + B increased formation of cornified envelopes. Similar irradiance doses of UVA 80 minutes (1.12 J/cm2) and UVA + B 40 minutes (1.04 J/cm2) caused 2.4- and 3.3-fold increases in cornified envelope formation, respectively. With solar-simulated irradiation, the cornified envelope formation was increased by 3.5-fold after exposure of 8 minutes (2.6 J/cm2). Irradiation of melanocytes with UVA, UVA + B, or solar-simulated irradiation resulted in a dose dependent decrease in melanocyte numbers after 24 hours compared with sham irradiated controls. As a result of UV irradiation, tyrosinase activity of melanocytes measured at 24 hours was stimulated. UVA + B irradiation (1.04 J/cm2) increased tyrosinase activity approximately twofold, while UVA alone (1.1 J/cm2) increased tyrosinase four to sixfold and solar-simulated irradiation (1.3 J/cm2) increased tyrosinase approximately twofold compared to the control cells. Melanin content increased in cells after both UVA and mixed UVA + B irradiation. These results indicate that both UVA and mixed UVA + B irradiation had qualitatively similar effects on the proliferative and functional activity of skin-derived cells but that the type of irradiation and the dosage regimen affect the dose response relationship. PMID- 8408232 TI - Bacterial lipopolysaccharide induces actin reorganization, intercellular gap formation, and endothelial barrier dysfunction in pulmonary vascular endothelial cells: concurrent F-actin depolymerization and new actin synthesis. AB - Bacterial lipopolysaccharide (LPS) influences pulmonary vascular endothelial barrier function in vitro. We studied whether LPS regulates endothelial barrier function through actin reorganization. Postconfluent bovine pulmonary artery endothelial cell monolayers were exposed to Escherichia coli 0111:B4 LPS 10 ng/ml or media for up to 6 h and evaluated for: 1) transendothelial 14C-albumin flux, 2) F-actin organization with fluorescence microscopy, 3) F-actin quantitation by spectrofluorometry, and 4) monomeric G-actin levels by the DNAse 1 inhibition assay. LPS induced increments in 14C-albumin flux (P < 0.001) and intercellular gap formation at > or = 2-6 h. During this same time period the endothelial F actin pool was not significantly changed compared to simultaneous media controls. Mean (+/- SE) G-actin (micrograms/mg total protein) was significantly (P < 0.002) increased compared to simultaneous media controls at 2, 4, and 6 h but not at 0.5 or 1 h. Prior F-actin stabilization with phallicidin protected against the LPS induced increments in G-actin (P = 0.040) as well as changes in barrier function (P < 0.0001). Prior protein synthesis inhibition unmasked an LPS-induced decrement in F-actin (P = 0.0044), blunted the G-actin increment (P = 0.010), and increased LPS-induced changes in endothelial barrier function (P < 0.0001). Therefore, LPS induces pulmonary vascular endothelial F-actin depolymerization, intercellular gap formation, and barrier dysfunction. Over the same time period, LPS increased total actin (P < 0.0001) and new actin synthesis (P = 0.0063) which may be a compensatory endothelial cell response to LPS-induced F-actin depolymerization. PMID- 8408233 TI - Use of p112-deficient myoblasts to determine the temporal order of the in vitro expression of myogenic components. AB - The present investigation examines the function and site(s) of involvement of an ecto-protein kinase and its substrate protein (a cell surface 112 kDa protein) in the in vitro myogenic pathway. The phosphorylated 112 kDa protein (p112) has recently been shown to be involved in myogenesis. Not much information is currently available on the role of the ecto-protein kinase and the 112 kDa protein in modulating the expression of the myogenic factors and various muscle specific proteins. Five different p112-deficient rat myoblasts were used to examine the temporal order of the in vitro expression of the myogenic components; namely, L6 myoblasts treated with BrdUrd or phloretin, a conditional p112 defective mutant (clone D1), an ecto-protein kinase-deficient mutant (clone F72), and a mutant defective in the 112 kDa protein (clone D1/S4). All these p112 deficient myoblasts were also impaired in myogenesis. The absence of p112, ecto protein kinase, and/or the 112 kDa protein was found to have no effect on the Myf 5 mRNA level. However, the expected increase in NCAM and Myf-4 mRNAs was not observed in any of the p112-deficient myoblasts examined. This suggests that the p112 site of action is probably located upstream of the Myf-4 and NCAM sites in the myogenic pathway. While 7-28 fold increases in the MLC, MHC, and TnT transcripts were observed during myogenesis, such increases were not observed in the p112-deficient myoblasts. However, when mutant D1/S4 was transfected with the myf-4 cDNA, expression of Myf-4 in the transfectant resulted in increased level of the MLC, MHC, and TnT mRNAs, and in myotube formation, even though the Myf-5 and NCAM mRNA levels and p112 were not altered. This suggests that p112 may function by activating transcription of Myf-4, which will subsequently promote the expression of muscle-specific proteins and myotube formation. In the absence of p112, Myf-5 cannot activate the expression of Myf-4, NCAM, MLC, MHC, TnT, and myotube formation. If all these components are involved in the same myogenic pathway, then p112 may be acting downstream from Myf-5, and upstream from NCAM and Myf-4. PMID- 8408234 TI - Evidence that a neutral cholesteryl ester hydrolase is responsible for the extralysosomal hydrolysis of high-density lipoprotein cholesteryl ester in rat hepatoma cells (Fu5AH). AB - Diethylumbelliferyl phosphate (UBP) has been shown to inhibit the neutral cholesteryl ester hydrolase activity responsible for hydrolysis of cellular lipid droplet cholesteryl ester (Harrison et al., 1990). The potential for (UBP) to inhibit uptake and hydrolysis of high density lipoprotein (HDL) cholesteryl ester was studied in Fu5AH hepatoma cells, a model for HDL cholesterol delivery. Coincubation of 3H-cholesteryl ester labeled HDL with UBP resulted in a 72% decrease in the cellular free cholesterol/cholesteryl ester (FC/CE) isotope ratio, indicating an inhibition in the conversion of cholesteryl ester to free cholesterol. Total cellular 3H-CE uptake was modestly (27%) but significantly decreased by UBP. Pulse-chase experiments (15 min. pulse and 7 min. chase) were used to study the hydrolysis of HDL 3H-CE in subcellular fractions separated by percoll gradients. The conversion of 3H-CE to 3H-FC could be demonstrated in fractions that comigrated with the plasma membrane/endosome fractions but were well separated from lysosomes. Neutral cholesteryl ester hydrolase activity was detected in those same fractions. These results suggest that an extralysosomal pathway is operating in the metabolism of HDL cholesterol and its delivery to hepatoma cells. PMID- 8408235 TI - Asymmetric origins of the mature glomerular basement membrane. AB - Renal plasma filtration is a critical physiologic function that depends upon the precise composition and arrangement of the constituent extracellular matrix proteins within the glomerular basement membrane (GBM). The GBM develops during renal embryogenesis by the fusion of discrete basement membranes produced independently by endothelial and visceral epithelial cells, and, possibly from matrix secreted by the mesangial cells. In the mature animal, however, the epithelial cell has generally been accepted as the sole source of all GBM constituent proteins. Although the final structures and distributions of the component proteins have been defined by histochemical techniques, the individual contributions of the three resident glomerular cell types to the maintenance and turnover of the mature GBM remain uncertain. We report the application of a new technique, in situ reverse transcription (ISRT), for the localization of RNA transcripts of nine major GBM protein components within the closely apposed cells of the glomerulus. Using this technique, we demonstrate that in normal adult rat glomeruli the RNA transcripts for heparan sulfate proteoglycan and the laminin-S chain are primarily expressed by visceral epithelial cells, while Type IV alpha-1 and alpha-2 collagen transcripts were restricted to the endothelial cells in a heterogeneous pattern. RNA transcripts for entactin and the laminin-A and -B2 chains were expressed by all three glomerular cell types, while laminin-B1 and fibronectin transcripts were limited to the mesangium. These findings demonstrate that GBM synthesis in the mature animal is not restricted to the epithelial cell and that all intrinsic glomerular cells contribute to the production of GBM protein components. The ISRT technique also provided the additional, and unexpected, finding that appreciable synthetic heterogeneity exists within individual glomerular cell types. PMID- 8408236 TI - Comparative studies of the granulopoietic enhancing effects of biosynthetic human insulin-like growth factors I and II. AB - The effect of biosynthetic human insulin-like growth factor I (IGF-I) and IGF-II on the in vitro growth of human marrow myeloid progenitors in the presence of recombinant human granulocyte colony stimulating factor (rhG-CSF), granulocyte macrophage CSF (rhGM-CSF), or interleukin-3 (rhIL-3), was investigated. IGF-I and IGF-II similarly enhanced the growth of myeloid progenitors in cultures stimulated with any of the above hemopoietic regulators. Analysis of colony composition showed an increase in the numbers of granulocyte colonies, but no alteration in the numbers of macrophage or granulocyte/macrophage colonies. IGF-I induced an increase of 62 +/- 16%, 84 +/- 13%, and 107 +/- 18% in granulocyte colony numbers in the presence of G-CSF, GM-CSF, or IL-3, respectively. The values for IGF-II were 66 +/- 13%, 96 +/- 12%, and 91 +/- 12%. Similar enhancement of myeloid colony formation by both peptides was also detected in G CSF and GM-CSF-stimulated cultures of marrow cells that had been depleted of accessory cells, while neither peptide exerted any effect in the presence of IL-3 in such cultures. The growth-promoting effects of IGF-I and IGF-II were completely abrogated by monoclonal antibodies directed against the IGF-I (Type I) membrane receptor. IGF-I and IGF-II thus appear to exert their effects on human marrow myeloid progenitors via a direct mechanism involving the Type I receptor. PMID- 8408237 TI - Anti-integrin antibodies induce type IV collagenase expression in keratinocytes. AB - During wound healing, pericellular proteolysis is thought to be essential for the detachment of keratinocytes from basement membrane and in their migration into the wound bed. We have characterized integrin-type cell adhesion/migration receptors in human mucosal keratinocytes and examined their function in the regulation of type IV collagenase gene expression. Two major integrins of the beta 1 class, alpha 2 beta 1 and alpha 3 beta 1, were found to function as collagen and fibronectin receptors, respectively. Antibodies against beta 1 and alpha 3 integrin subunits were found to stimulate the expression of the 92 kDa type IV collagenase severalfold in a dose-dependent manner. Keratinocytes expressed also the 72 kDa type IV collagenase, the synthesis of which remained, however, unchanged in keratinocytes treated with anti-integrin antibodies. Stimulation of 92 kDa enzyme was found to be caused directly by antibody binding to integrins, since Fab-fragments of anti-beta 1 antibodies alone were able to induce collagenase expression in the absence of secondary, clustering antibodies. Antibodies against alpha 2 beta 1 integrin caused no stimulation. Keratinocytes seeded on different substrata (plastic, collagen, fibronectin, laminin, or vitronectin) showed equal induction of type IV collagenase expression. Expression of 92 kDa type IV collagenase could not be induced by peptides (GRGDS, GRGES), proteins (fibronectin, laminin, fibrinogen, albumin), or antibodies to fibronectin. We suggest that proteolytic processes around keratinocytes can be regulated by extracellular factors signalling through integrin-type receptors. PMID- 8408238 TI - Role of glucose in interleukin-1 beta production by lipopolysaccharide-activated human monocytes. AB - When monocytes are activated with endotoxin (lipopolysaccharide [LPS]), they make and release several mediators, including interleukin-1 beta (IL-1 beta). This study was undertaken to investigate the role of glucose in IL-1 beta production by these cells. IL-1 beta was produced in a dose-dependent manner to glucose concentration in the culture medium. The uptake of (3H)2-deoxyglucose in monocytes was stimulated by LPS 1,554% after 10 minutes, 6,095% after 2 hours, then gradually declined after 4 hours of incubation. The inhibition of the uptake of (3H)2-deoxyglucose by either 10 microM cytochalasin B or phloretin, added at the time of monocyte activation, was accompanied by significant reduction in ATP/ADP ratio and the inhibition of the production of IL-1 beta by activated monocytes. The synthesis of total protein did not change in monocytes activated in the absence of glucose in the culture medium, nor in the presence of either 10 microM cytochalasin B or phloretin. The export of IL-1 beta from LPS-activated monocytes was not inhibited by either 10 microM cytochalasin B or phloretin, nor in the absence of glucose in the culture medium. These data suggest that 1) glucose is required for LPS-induced IL-1 beta production by monocytes; 2) glucose is the major source of ATP for IL-1 beta production; 3) glucose transporter (GLUT 1) does not control the export of IL-1 beta. PMID- 8408239 TI - Thrombospondin mediates migration and potentiates platelet-derived growth factor dependent migration of calf pulmonary artery smooth muscle cells. AB - A precipitating factor in the development of atherosclerotic lesions is the inappropriate migration and proliferation of vascular smooth muscle cells (SMC) within the intima of the vessel wall. Focusing on the role of extracellular matrix proteins in SMC migration, we have demonstrated that thrombospondin (TSP) itself is a potent modulator of SMC motility and acts to potentiate platelet derived growth factor (PDGF)-mediated SMC migration as well. Migration of SMC to TSP was dose dependent. Interestingly, maximal SMC migration to TSP exceeded that to either PDGF or basic fibroblast growth factor (bFGF). The distal COOH terminus of TSP was shown to mediate SMC migration as demonstrated by complete inhibition of the response by monoclonal antibody (mAb) C6.7. Nevertheless, proteolytic fragments of TSP were not as potent as intact TSP in mediating SMC migration. Only by combining the heparin-binding domain (HBD) with the 140 kD COOH terminal fragment was SMC migration restored to levels seen with intact TSP. Based on antibody inhibition studies, an alpha v-containing integrin receptor, but not alpha v beta 1 or alpha v beta 3, appeared to be involved in SMC migration to TSP. The coincidental expression of PDGF and TSP at sites of vascular injury and inflammation led us to evaluate the effect of suboptimal levels of TSP on SMC responsiveness to PDGF. SMC migration in response to PDGF was enhanced nearly 60% in the presence of suboptimal concentrations of TSP. This effect was specific for PDGF and dependent on the concentration of TSP with maximal potentiation obtained between 50-100 nM TSP, concentrations tenfold lower than those necessary for SMC migration to TSP itself. mAb C6.7 completely inhibited enhancement but, as with SMC migration to TSP alone, TSP proteolytic fragments did not possess the effectiveness of the intact molecule. Additional experiments assessing SMC migration to PDGF demonstrated that PDGF stimulated SMC motility indirectly by inducing TSP synthesis. These studies suggested that TSP functions as an autocrine motility factor to modulate SMC migration, which in conjunction with PDGF could serve to aggravate and accelerate development of atherosclerotic lesions at sites of vascular injury or inflammation. PMID- 8408240 TI - Growth promotion of transfected hepatoma cells by liver fatty acid binding protein. AB - Former studies have linked hepatocyte growth with liver fatty acid binding protein (L-FABP) of rat liver cytosol. In search for the roles of L-FABP in hepatocytes, we previously stably transfected rat L-FABP sense and antisense cDNAs into rat hepatoma HTC cells that do not contain L-FABP RNA or protein, thereby providing a zero-background, homologous cell model of L-FABP-expression suitable for controlled studies of its intracellular functions in hepatocyte derived cells. The present study demonstrates the abilities of L-FABP to promote DNA synthesis and cell growth, preserve cell morphology, extend survival, and act cooperatively with unsaturated fatty acids in the transfected hepatoma cells in the absence of serum. Following removal of serum, the three control L-FABP nonexpressing cell lines increased in cell lines increased in cell number for 24 hr and thereafter declined, whereas the three L-FABP-expressing cell lines exhibited a 39% higher rate of DNA synthesis per cell at 24 hr and grew in cell number for 48 hr. As a result, at 72 hr there were 2.5-fold (avg.) as many L-FABP expressing cells than L-FABP-nonexpressing cells. In addition, the L-FABP expressing cells retained their original polygonal morphology at 48 hr, when in contrast most of the control nonexpressing cells were spherical in shape with membrane blebs. In an effort to identify the agonists that collaborate with L FABP in the growth promotion and preservation of cell morphology, various free fatty acids were examined at 48 hr for their ability to eliminate the differences in behavior of the two cell types in the serum-free medium. The unsaturated fatty acids, oleic acid (18:1 omega 9), linoleic acid (18:2 omega 6), alpha-linolenic acid (18:3 omega 3), and arachidonic acid (20:4 omega 6), at 1 microM markedly elevated the level of DNA synthesis in the more depressed control L-FABP nonexpressing cells and moderately raised it in the less depressed L-FABP expressing cells. In accord, the control L-FABP-nonexpressing cells needed 10(-6) 10(-5) M linoleic acid to achieve the extent of DNA synthesis attained by the expressing cells in the absence of added fatty acid. At 10 microM linoleic acid, their levels of DNA synthesis were equal. In contrast, five saturated fatty acids had no detectable effect on DNA synthesis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8408241 TI - Cell adhesion markers are expressed by a stable human endothelial cell line transformed by the SV40 large T antigen under vimentin promoter control. AB - Markers of endothelium have been studied in a new endothelial cell line derived from human umbilical cord vein cells by microinjection of a recombinant gene that includes a deletion mutant of the human vimentin gene regulatory region controlling the large T and small t antigen coding region of the SV40 virus. In culture, this immortalized venous endothelial cell line (IVEC) demonstrated morphological characteristics of endothelium; uptake of acetylated low density lipoprotein and presence of the Factor VIII-related antigen. Treatment of IVEC cells with Interleukin-1 beta (IL-1 beta) at 10 U.ml-1 activates the expression of cell adhesion molecules such as endothelial leucocyte adhesion molecule (ELAM 1), intercellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1), as observed in primary culture. Prostacyclin secretion was induced in the IVEC cells by 100 nM PMA treatment and thrombin at 0.5 U/ml. Angiotensin converting enzyme (ACE) activity detected in IVEC cells was present but lower than ACE activity in primary endothelial cells and was completely blocked by enalaprilat (1 microM), a specific ACE inhibitor. The presence of ACE mRNA was also demonstrated in IVEC cells by RT-PCR amplification. Our data demonstrate that endothelial cells immortalized by use of this recombinant gene retain the morphological organization and numerous differentiated properties of endothelium. PMID- 8408242 TI - Constitutive expression of c-myc does not relieve cAMP-mediated growth arrest in human lymphoid Reh cells. AB - The Reh cell system is suitable for evaluating events important for control of proliferation independently of mechanisms involved in differentiation, as Reh cells are unable to differentiate. In the human pre-B cell line Reh, activation of adenylate cyclase by forskolin induces a five to tenfold rapid, transient down regulation of steady-state c-myc RNA within 4 hours. Concurrently, the cells are strongly growth arrested in the G1 phase of the cell cycle. To clarify if the observed growth arrest could be relieved by constitutive expression of c-myc, an exogenous c-myc gene under constitutive promoter control was introduced into Reh cells by electroporation. The c-myc-expressing construct pDMmycHyg contained human c-myc exons 2 and 3 driven by the Mo-MLV LTR and conferred hygromycin resistance. Exogenous c-myc RNA transcripts and protein were constitutively expressed in the transfected clones at levels roughly twice as high as the level in nontransfected cells. Total c-myc protein levels were unchanged upon treatment of transfected clones with forskolin. Yet, the transfected cells were not released from growth arrest. Furthermore, the transfected Reh cells did not differentiate upon forskolin treatment. Constitutive overexpression of c-myc is therefore not sufficient for relieving forskolin-mediated effects on growth arrest in Reh cells. PMID- 8408243 TI - Transcriptional and posttranscriptional regulation of hepatic beta 2-adrenergic receptor gene expression during development. AB - Hepatic responsiveness to beta 2-adrenergic stimulation is dynamically regulated during early development as well as following hepatic injury and disease. In the present study, the molecular mechanisms that underlie the decline in the steady state levels of hepatic beta 2-adrenergic receptor mRNA that occurs during development in the male rat were investigated. As determined by nuclear run-on assays, an age-associated reduction in beta 2-adrenergic receptor gene transcription was observed. The transcription rate of the beta 2-adrenergic receptor gene in postnatal day 18 liver was approximately 50% lower than that of fetal liver. Stability of beta 2-adrenergic receptor gene transcripts was highest (t1/2 approximately 6h) in hepatocytes isolated from fetal rats and was lowest (t1/2 approximately 6h) in hepatocytes from postnatal day 14 rats. In fetal hepatocytes, but not postnatal day 2 hepatocytes, cycloheximide appeared to stabilize beta 2-adrenergic receptor gene transcripts in the presence of actinomycin D. These findings establish the molecular basis of reduced steady state levels of beta 2-adrenergic receptor mRNA in liver during early postnatal development and suggest multilevel regulatory control of hepatic beta 2 adrenergic receptor gene expression. PMID- 8408244 TI - Adenine nucleotides and adenosine metabolism in pig kidney proximal tubule membranes. AB - Exogenous adenosine triphosphate (ATP) added to brush-border membrane vesicles was rapidly degraded mainly to inosine according to the high ecto-nucleotidase activities in these vesicles. In the absence of phosphate, inosine was slowly transformed into hypoxanthine, and xanthine oxidase and dehydrogenase activities were not detected. The presence of ecto-adenosine deaminase and ecto-adenosine monophosphate (AMP) nucleotidase was shown. The ecto-adenosine deaminase was inhibited by deoxycoformycin and was also detected in rat renal brush-border membrane vesicles. Using orthovanadate, levamisole, and alpha, beta-methylene adenosine diphosphate as possible inhibitors, alkaline phosphatase was shown to be the main agent responsible for ecto-AMP nucleotidase activity. In pig renal basolateral membrane vesicles and in whole cell extracts from pig renal cortex, ecto-AMP nucleotidase was the limiting factor in ATP degradation. Comparing the ATP catabolism in the whole cell cortical extract with the catabolism in the same sample precleared of membranes, it was shown that ectonucleotidase activity is mainly bound to the membranous components. It is also shown that the whole cell extract of pig renal cortex has hypoxanthine phosphoribosyl transferase activity, and it seems probable that the rapid and specific formation of luminal inosine and its transport into the cell in competition with adenosine may start the purine salvage pathway through the synthesis of IMP from hypoxanthine. PMID- 8408245 TI - Leukocyte-deactivating factor from macrophages: partial purification and biochemical characterization. A novel cytokine. AB - A deactivating factor (MDF) is released from granuloma-like lesions of mice (giant and epithelioid macrophages) to the surrounding medium. Test cells incubated in the presence of MDF display dramatic inhibition of superoxide anion (O2-) release when stimulated. This failure to manifest O2 release is observed whether PMA, all-transretinal, or fMet-Leu-Phe is the stimulating agent. MDF acts on different cell types from different species; mouse macrophages as well as guinea pig, human, and mouse neutrophils. Such results suggest that it is a universal regulatory cytokine with high affinity for phagocytic lineages. The factor was subjected to various purification methods: ultrafiltration, gel chromatography, and reversed phase HPLC. A crude preparation that resulted from conditioning of medium by old macrophages (MCM) shows two peaks of activity when subjected to gel filtration. These correspond to molecular weights for the active principle of 3 and 11 kD. When the factor was obtained by extraction of the same cells after washing and sonication, only the former peak was seen. Fractions corresponding to a MW of 3 kD from several preparations were combined and subjected to HPLC. MDF activity then appeared in a single fraction. MDF is thus putatively a modulator of the cidal activity of phagocytic cells that utilize release of reactive oxygen species for cytocidal activity. PMID- 8408246 TI - Down-regulation of retinoic acid receptor activity associated with decreased alpha and gamma isoforms expression in F9 embryonal carcinoma cells differentiated by retinoic acid. AB - F9 embryonal carcinoma cells differentiate in response to retinoic acid (RA). To investigate the regulation of RA receptors (RARs) expression during this process, cDNA probes specific for the major RAR isoforms were used. In contrast to the level of RAR beta 2 mRNA which was high in cells treated 5 days with RA and below detection in untreated cells, as previously described, the steady state levels of RAR alpha 1, alpha 2, gamma 1, and gamma 2 mRNAs were markedly decreased in the RA-differentiated cells as compared to untreated cells. The down-regulation of the RA-responsive system in differentiated cells was also evident in gel shift assays as a marked decrease in binding capacity to a retinoid acid response element (beta 2RARE), as well as in chloramphenicol acetyltransferase (CAT) assays as a sixfold decrease in RA-mediated transacting activity via this element. The down-regulation of RAR DNA-binding and transacting activity coincided with the burst in tissue plasminogen activator secretion and thus, occurred at the hinge between early and late differentiation. The down-regulation of RA responsiveness may constitute an important event in the transition between early and late differentiation stage in F9 cells. PMID- 8408247 TI - Density-dependent inhibitory effect of transforming growth factor-beta 1 on human fibroblasts involves the down-regulation of platelet-derived growth factor alpha receptors. AB - We have previously found that transforming growth factor-beta 1 (TGF-beta 1) inhibits the mitogenic activity of platelet-derived growth factor (PDGF) in cultures of human neonatal fibroblasts in a density-dependent fashion. In the present investigation we determined the effect of TGF-beta 1 on the PDGF alpha receptor, which binds all PDGF isoforms, as well as on the beta-receptor, which binds only PDGF-BB with high affinity. We found that the inhibitory effect of TGF beta 1 on PDGF-AA-induced mitogenesis was density-dependent; when dense cell cultures were preincubated with TGF-beta 1, there was an complete inhibition of 3H-thymidine incorporation, whereas the effect was less in sparse cultures. A similar density-dependent effect of TGF-beta 1 was seen in PDGF-BB treated cells, although less pronounced. The binding of 125I-labeled PDGF-AA and PDGF-BB to the alpha-receptor was significantly reduced after treatment with TGF-beta 1 in dense cultures, whereas the sparse cultures were less affected. A decrease of alpha receptor mRNA was also seen. The levels of beta-receptor protein and mRNA were unaffected. We conclude that the growth inhibitory effect of TGF-beta 1 is cell density-dependent and involves down-regulation of PDGF alpha-receptors. PMID- 8408248 TI - The history of Kienbock's disease. AB - The history of Kienbock's disease is outlined from its description in 1910 in Europe, to its acceptance as a disease entity in the United States, to the advent of therapeutic surgical treatment in 1950. The concepts developed during this period, such as the association of ulnar variance and Kienbock's disease, are the basis for many of the present day surgical interventions performed. PMID- 8408249 TI - Vascularity of the lunate. Anatomic studies and implications for the development of osteonecrosis. AB - Osteonecrosis of the lunate is well established as the pathologic entity underlying the clinical condition known as Kienbock's disease. Consequently, the extraosseous and intraosseous blood supplies to the lunate have been well studied. A series of three palmar and three dorsal arterial arches provide a consistent volar supply to the lunate, and a frequent, but inconsistent, dorsal supply. Volar and dorsal foramina each generally contribute one to two vessels to a volar and dorsal intraosseous arterial network. The dorsal and volar arterial systems anastomose distal to the midline of the lunate; however, in as many as 7.5% of lunates, no anastomosis my be present. Between 7% and 26% of lunates may lack either a volar or dorsal arterial supply. These findings have significant implications regarding possible mechanisms for the development of osteonecrosis and the treatment of Kienbock's disease. PMID- 8408250 TI - Diagnosis and clinical findings of Kienbock's disease. AB - The clinical presentation and history of patients with Kienbock's disease do not permit differentiating avascular necrosis of the lunate from vague wrist pathology of a variety of causes. Diagnosis and evaluation of this disorder therefore require appropriate imaging studies. Plain-film radiography, tomography, and bone scintigraphy are useful screening tests, but MRI yields a definitive diagnosis. CT is also valuable for delineating structural changes and the amount of collapse in the lunate. The staging of Kienbock's disease is based on clinical and radiographic data. PMID- 8408251 TI - Staging and its use in the determination of treatment modalities for Kienbock's disease. AB - For more than 80 years, surgeons have staged an unsuccessful search for a universally acceptable treatment for Kienbock's disease. It is our contention that no single treatment will be universally successful. Treatment choice must be based on a number of variables, including the experience of the surgeon, the desires and activity level of the patient, the anatomic variation of the ulna, and most importantly, on the stage of the disease. In the early stages, efforts should be made to salvage the lunate and prevent loss of normal architecture. In the later stages, efforts should be made to restore that architecture. In end stage, normal architecture must be sacrificed to restore function. Currently we recommend immobilization with possible equalization procedures for patients with ulnar-minus variance and stage I disease. In a patient with stage I disease and ulnar-positive variance, we recommend immobilization with consideration for a revascularization procedure. For stages II or IIIA disease with ulnar-minus variance, we attempt an equalization procedure. For stages II or IIIA disease with ulnar-positive variance, we recommend revascularization as performed by Hori. In stage IIIB disease, we prefer a triscaphe fusion to restore carpal stability and prevent further degeneration. In stage IV disease, proximal row carpectomy or wrist arthrodesis is indicated. PMID- 8408252 TI - Biomechanical comparison of methods used to treat Kienbock's disease. AB - The surgical approach to Kienbock's disease is largely dependent on the stage of the disease and the ulnar variance pattern. Many of the surgical treatments are designed to unload the lunate, halt disease progression, and allow for possible revascularization. This article reviews a collection of studies investigating the biomechanical effects of load-altering procedures. Knowledge of the biomechanical impact of the various operative interventions is clinically useful in creating a treatment algorithm. PMID- 8408253 TI - Biomechanical evaluation of operative procedures to treat Kienbock's disease. AB - Operative procedures used to treat Kienbock's disease have been biomechanically evaluated experimentally. We have shown that joint leveling procedures, such as radial shortening and ulnar lengthening, experimentally unload the ulna and the radial lunate fossa. For wrists with neutral ulnar variance, a lateral opening or medial closing radial wedge procedure unloads the radial lunate fossa. Scapho trapezio-trapezoidal fusion and scapho-capitate fusion also unload the radial lunate fossa but at the expense of loading the adjacent joints. Neither a capitate-hamate fusion nor a carpal tunnel release alter the radial ulnar carpal joint loading. PMID- 8408254 TI - The effect of force transmission on the carpus after procedures used to treat Kienbock's disease. AB - Analytic and experimental methods used for the study of force transmission through the carpus are summarized. Data on the force and pressure distribution under normal wrist conditions and after procedures used to treat Kienbock's disease are reviewed. Potential biomechanical factors causing the disease and possible directions of future studies are discussed. PMID- 8408255 TI - Revascularization procedures in the treatment of Kienbock's disease. AB - Lunate revascularization by means of vascular bundle implantation is one of the useful procedures for stages I and II Kienbock's disease, but for stage III, it should be combined with some biomechanical procedure or partial carpal fusion. There is no indication for this procedure in stage IV disease. PMID- 8408256 TI - Ulnar lengthening procedures. AB - Ulnar lengthening redistributes axial compressive forces partially relieving those on the lunate, slowing or halting further progressive collapse. Long-term results are satisfactory, although restitution of the carpal anatomy is not obtained. PMID- 8408257 TI - Radial shortening. AB - The association of a negative ulnar variance in individuals with Kienbock's disease provides the basic rationale for a radial shortening osteotomy in the treatment of this particular condition. Many biomechanical studies have demonstrated both high compressive and tensile forces on the lunate during normal ranges of wrist motion. An osteotomy to realign the radiocarpal joint by shortening the radius attempts to lessen the presumably increased compressive forces on the lunate in patients who have a negative ulnar variance. By "unloading" the lunate through this procedure, the possibility for secondary revascularization exists. It appears that the lunate "stands still in time" after a radial shortening, with no significant further deterioration being noted by objective measures. Nevertheless, evidence to concretely support revascularization is sketchy and subjective. The radiographic observation of lunate collapse (stage III disease) does not, by itself, represent a contraindication to the use of a radial shortening. In fact, excellent symptomatic relief with improvement in objective parameters is noted in patients with stage III disease. A distinct advantage of radial shortening, in all stages of disease, is the remote nature of the procedure itself, allowing quick postoperative recovery and the ability to undertake more complex carpal reconstructive procedures should the need arise. Radial shortening represents a relatively simple extra-articular procedure with excellent relief of pain, increased postoperative grip strength, and postoperative improvement of range of motion. Although some subjective evidence of revascularization of the lunate has been presented by several investigators, it does not appear that the external architecture of the lunate (be it collapsed or of normal height) improves or deteriorates with time after a radial shortening.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408258 TI - Implant resection arthroplasty in the treatment of Kienbock's disease. AB - The lunate implant acts as an articulating spacer to maintain the carpal bones relationship after a resection procedure. Its use helps preserve functional wrist mobility and stability. Our experience spans 25 years and the use of three materials. Since 1986, we have used lunate implants made of unalloyed titanium with very promising results. This procedure requires strict surgical indications, an exacting technique, associated intercarpal bone fusions as indicated, and the avoidance of abusive activity. PMID- 8408259 TI - Use of triscaphe fusion in the treatment of Kienbock's disease. AB - The disability of Kienbock's disease is a manifestation of carpal collapse. Significant loading for any wrist must be taken on the radial side. Triscaphe arthrodesis provides a support mechanism for the collapsed lunate, a solid bone cartilage mechanism for load transference to the radius, and maintenance of satisfactory motion. PMID- 8408260 TI - Scaphocapitate fusion in the treatment of Kienbock's disease. AB - The primary treatment goal for Kienbock's disease remains decompression of the lunate. Although a number of treatment options are available, scaphocapitate fusion is our first choice in many cases of Kienbock's disease. Scaphocapitate fusion mechanically decompresses the lunate and prevents progressive carpal instability. When compared with STT fusion, it has been shown to result in equal or relatively less loss of wrist motion and thumb basilar joint motion. For disease in earlier stages and with ulnar-minus variance, joint leveling procedures have been shown effective; but ulnar-neutral and ulnar-positive wrists would cause one to favor scaphocapitate fusion to prevent postoperative ulnocarpal abutment. PMID- 8408261 TI - Capitate shortening in the treatment of Kienbock's disease. AB - The goal of the technically simple procedure of capitate shortening with capitate hamate fusion is revascularization of the lunate, and on radiographic evaluation, the results have been satisfactory (Figs. 9 through 11). This surgical procedure offers the advantage of direct vision and, therefore, direct staging of the aseptic process. The procedure is designed for patients with early Kienbock's disease who have relatively minor architectural changes in the aseptic lunate, no arthritic changes, and no ulnar-minus variance. When these criteria are met, the clinical results have been encouraging and lasting. PMID- 8408262 TI - Excision and fascial interposition arthroplasty in the treatment of Kienbock's disease. AB - Capitate shortening with capitate-hamate fusion is a joint leveling procedure designed for patients with Kienbock's disease who are in the early phases of this disease; that is, they have minimal or no fragmentation of the lunate, no arthritis, and no ulnar-minus variance. The long-term clinical results have been very good, and revascularization occurs in a high percentage of cases. PMID- 8408263 TI - Wrist denervation procedures in the treatment of Kienbock's disease. AB - In almost all papers on wrist denervation, indications for denervation in Kienbock's disease has been mentioned without exact data. A detailed analysis of our clinical data on 61 patients proved the positive effect with respect to pain relief. In the 47 patients in which the denervation was combined with other procedures, such as radial shortening, ulnar lengthening, lunate excision, and replacement by silicone or tendon, pedicled pisiform transposition, and STT arthrodesis, a very high rate of patients (76%) had no complaints or pain only with heavy manual work. Of these treated patients, 88% were very or remarkably satisfied. About the same success rate was found in cases in which the denervation was applied as a single procedure. The extent of denervation was different; therefore, it can be concluded that partial or complete denervation has its value as an additional or single procedure in the treatment of Kienbock's disease, especially in the advanced stages (Decoulx's stages III and IV). PMID- 8408264 TI - "Salvage" procedures in the treatment of Kienbock's disease. Proximal row carpectomy and total wrist arthrodesis. AB - In symptomatic stage III Kienbock's disease, proximal row carpectomy may be considered not only as a secondary "salvage," but also a primary reconstructive procedure. The need for concomitant radial styloidectomy is assessed intraoperatively after removal of the proximal row. Distraction resection arthroplasty is an alternative for patients with full-thickness cartilage loss on the capitate head or in the lunate fossa surpassing 3 mm in diameter. For patients with generalized carpal arthrosis (stage IV Kienbock's disease) or unsuccessful previous reconstructive surgeries, total wrist arthrodesis is the most reliable procedure for pain relief. PRC is an acceptable alternative to TWA for stage IV disease if the patient is willing to risk having some residual pain to preserve wrist motion. Transection of the articular branch of the posterior interosseous nerve may be a useful adjunct to these procedures. PMID- 8408265 TI - The isolation and culture of microvascular endothelium. PMID- 8408266 TI - Desmoyokin, a 680 kDa keratinocyte plasma membrane-associated protein, is homologous to the protein encoded by human gene AHNAK. AB - We have obtained a monoclonal antibody (33A-3D) that specifically recognize desmoyokin, a 680 kDa desmosomal plaque protein that is well characterized in bovine muzzle epidermis. A cDNA clone (DY6, 3693 bp) was isolated by immunoscreening a mouse keratinocyte expression library with 33A-3D, and it was confirmed that DY6 has a partial coding sequence for desmoyokin. DY6 consists of highly homologous repeats about 128 residues long. Furthermore, the 128-residue repeats exhibit a quasi seven-residue substructure, which we believe will adopt an antiparallel beta-sheet structure. Surprisingly, the amino acid sequence showed a significant homology with AHNAK, a newly identified human gene encoding a 700 kDa protein, which was suggested to be down-regulated in neuroblastoma. From its extensive homology, the similarity in both size and structure, and the identical patterns on Southern blot analysis of genomic DNAs, desmoyokin and AHNAK protein are thought to be identical. Although the desmoyokin/AHNAK protein is detected in a variety of cell types at both protein and mRNA levels, its distribution in keratinocytes (associated closely with cell membrane) is quite different from that in cells other than keratinocytes (distributed diffusely in the cytoplasm). These findings suggest that the desmoyokin/AHNAK protein is a ubiquitous molecule with a unique structure and appears to have different distributions (and probably different functions) among different cells. PMID- 8408267 TI - Polarized distribution of Na,K-ATPase in honeybee photoreceptors is maintained by interaction with glial cells. AB - Arthropod photoreceptors are polarized cells displaying distinct surface domains. The distribution of the Na,K-ATPase (sodium pump) over these domains was examined in the honeybee photoreceptor using a monoclonal antibody that specifically recognizes the sodium pump alpha-subunit (approximately 100 kDa). We find that the sodium pump is restricted to sites of the nonreceptive photoreceptor surface closely juxtaposed to glial cells; no sodium pumps were detected on the glia-free domains of the nonreceptive surface and on the light-sensitive microvillar membranes. In order to determine the role of photoreceptor-glia contact in maintaining this polarized pump distribution, we assayed the distribution of the Na,K-ATPase after experimentally influencing photoreceptor-glia contact. Sodium pumps were present on the entire nonreceptive photoreceptor surface when photoreceptor-glia contact was removed by isolating the photoreceptors. Remodeling photoreceptor-glia contact by incubation in hyperosmotic saline caused a redistribution of sodium pumps on the photoreceptor surface corresponding to the redistribution of glial cells. We show, further, that both photoreceptor-glia contact and Na,K-ATPase distribution are independent of extracellular Ca2+. No junctional structures were observed at the borders between Na,K-ATPase-positive and Na,K-ATPase-negative membrane domains. Together, these results suggest that adhesion of glial cells to the photoreceptors plays a crucial role in the maintenance of the polarized distribution of Na,K-ATPase in the honeybee photoreceptors. The Ca(2+)-independent adhesion of glial cells to the photoreceptor surface may trap the pump molecules at the sites of photoreceptor glia contact. PMID- 8408268 TI - A methodology for the systematic and quantitative study of cell contact guidance in oriented collagen gels. Correlation of fibroblast orientation and gel birefringence. AB - Despite the likely role of contact guidance in every physiological process involving cell migration, its study in a three-dimensional tissue-equivalent environment has been precluded, heretofore, by inherent difficulties in systematically preparing well-defined contact guidance fields and quantifying the resultant contact guidance. Here, we describe a novel use of a magnetic field to orient collagen fibrils during fibrillogenesis, entrapping cells dispersed in the collagen solution. Using computer-controlled staging and image analysis, we show from automated birefringence measurements of the resultant slab of cell-populated gel contained in a specially designed observation chamber that the fibril orientation is biased along the long axis of the chamber uniformly throughout the chamber. Further, we show that the degree of fibril orientation, and consequently the elicited contact guidance, can be controlled by independently varying the magnetic field strength or temperature during fibrillogenesis. We characterize the contact guidance response to the imposed contact guidance field by measuring cell orientation relative to the axis of fibril orientation from still images obtained in time-lapse via automated image analysis. We present the first quantitative correlation of contact guidance (based on cell orientation) with collagen fibril orientation (based on birefringence) for human foreskin fibroblasts cultured in a collagen gel, by using gels of varying orientation resulting from different magnetic field strengths and temperatures during fibrillogenesis, and by using sufficiently low cell concentrations and early observation times. PMID- 8408269 TI - Distribution of snRNPs, splicing factor SC-35 and actin in interphase nuclei: immunocytochemical evidence for differential distribution during changes in functional states. AB - Small nuclear ribonucleoproteins (snRNPs) play an integral role in the processing of pre-mRNA in eukaryotic nuclei. snRNPs often occur in a speckled intranuclear distribution, together with the non-snRNP splicing factor SC-35. snRNPs have also been shown to be associated with actin in the nuclear matrix, suggesting that both actin and snRNPs may be involved in the processing and transport of transcripts. The work reported here was undertaken to compare the spatial relationship of snRNPs, SC-35, and intranuclear actin in neuronal and non neuronal cell types. In undifferentiated PC12 cells and in non-neuronal cells growing in association with dorsal root ganglion neurons, confocal immunocytochemistry revealed a typical, speckled distribution of snRNP aggregates, which colocalized with the SC-35 splicing factor. In contrast, a unique snRNP distribution was observed in dorsal root ganglion neurons in vitro and in PC12 cells differentiated by nerve growth factor. In nuclei of these cells, snRNPs were predominantly located at the periphery where they formed a spherical shell apposed to the nuclear envelope. Ultrastructural immunogold labelling of snRNPs in dorsal root ganglion neurons in vitro confirmed this distribution. In contrast, SC-35 remained distributed in a speckled pattern throughout nuclei of dorsal root ganglion neurons and PC12 cells, even in cases where snRNPs were almost exclusively positioned at the nuclear periphery. In non neuronal cells in dorsal root ganglion cultures and in undifferentiated PC12 cells, snRNP aggregates were frequently associated with actin aggregates, as determined by Nearest Neighbor Analyses. In PC12 cells, this spatial relationship was altered during nerve growth factor-induced differentiation, prior to the time at which these cells showed morphological evidence of differentiation. Specifically, Nearest Neighbor Analyses between snRNP and actin aggregates in PC12 cells exposed to nerve growth factor for 4 hours revealed that snRNP and actin aggregates exhibited a closer association than in undifferentiated cells. These results suggest that sites of pre-mRNA processing and transcription may differ between cell types, and that the functions of snRNPs and actin within interphase nuclei may be related. The results also indicate that the distribution of snRNPs is dynamic and that it may depend upon the functional state of the cell as well as upon its state of differentiation. PMID- 8408270 TI - Centromeric DNA cloned from functional kinetochore fragments in mitotic cells with unreplicated genomes. AB - Treatment of cells arrested in the cell cycle at the G1/S-phase boundary with 5 mM caffeine induces premature mitosis, resulting in chromosomal fragmentation and detachment of centromere-kinetochore fragments, which are subsequently attached to the mitotic spindle and segregated in anaphase. Taking advantage of this in vivo separation of the centromere, we have developed a procedure for isolation of a centromere-enriched fraction of mitotic chromatin. Using this method, we have isolated and cloned DNA from the centromere-enriched material of Chinese hamster cells. One of the clones thus obtained was characterized in detail. It contains 6 kb of centromere-associated sequence that exhibits no recognizable homology with other mammalian centromeric sequences and is devoid of any extensive repetitive structure. This sequence is present in a single copy on chromosome 1 and is species-specific. Distinctive features of the clone include the presence of several A+T-rich regions and clusters of multiple topoisomerase II consensus cleavage sites and other sequence motifs characteristic of nuclear matrix associated regions. We hypothesize that these features might be related to the more compact packaging of centromeric chromatin in interphase nuclei and mitotic chromosomes. PMID- 8408271 TI - Transepithelial calcium transport in the chick chorioallantoic membrane. I. Isolation and characterization of chorionic ectoderm cells. AB - The chicken eggshell supplies approximately 80% of the calcium found in the hatchling chick. The mobilization of eggshell calcium into the developing embryo involves the transepithelial transport of large amounts of calcium in a development-specific manner. The cells responsible for the transport of eggshell calcium into the embryonic circulation are the ectodermal cells of the chorioallantoic membrane. In this report, we present a method for the isolation and culture of chorioallantoic membrane ectodermal cells, which are amenable to direct experimental manipulation. Cell preparations are characterized with respect to the expression of an ectoderm-specific cell surface marker (transcalcin, a calcium-binding protein), and a specific enzymatic activity (elevated Ca(2+)-activated ATPase). Functional assessment of in vitro cellular calcium uptake by 45Ca2+ tracer kinetics indicates the persistence of a temperature-sensitive, rapid-influx pathway similar to that observed in vivo. The preparations of primary ectodermal cells present an in vitro system applicable to the experimental analysis of calcium metabolism and transport by the chick chorioallantoic membrane. PMID- 8408272 TI - Transepithelial calcium transport in the chick chorioallantoic membrane. II. Compartmentalization of calcium during uptake. AB - Calcium transport from the eggshell to the developing chick embryo is carried out by the ectoderm cells of the chick chorioallantoic membrane. Primary cells isolated from chick chorioallantoic membrane ectoderm were used to analyze the subcellular distribution of 45Ca2+ accumulated from the extracellular medium. We present evidence suggesting that calcium may be sequestered into endosome-like vesicles during the initial phase of uptake. A combination of techniques were utilized to monitor calcium fluxes and calcium compartmentalization in the cultured chorioallantoic membrane cells: (1) fura-2 fluorescence was used to indicate cytosolic free calcium concentrations, (2) 45Ca2+ tracer was used to follow calcium accumulation in all cellular compartments, and (3) digitonin was used to differentially permeabilize subcellular membranes in order to localize 45Ca2+ by following tracer release profiles. Differences between cytosolic calcium flux and whole cell calcium accumulation suggested that the pathway of calcium uptake from the medium involves sequestration into an internal compartment separate from the cytosol. Kinetic analysis of the digitonin-mediated release of specific subcellular markers (lactate dehydrogenase, NAD-dependent isocitrate dehydrogenase, [3H]inulin, and [3H]-2-deoxyglucose) and preloaded 45Ca2+ indicated that calcium was localized in a compartment similar to endosomal vesicles. Our results are consistent with a transcytotic mechanism for chorioallantoic membrane calcium transport. PMID- 8408273 TI - Xenopus nuclear factor 7 (xnf7) possesses an NLS that functions efficiently in both oocytes and embryos. AB - Xenopus nuclear factor 7 (xnf7) is a nuclear phosphoprotein that is encoded by a member of a novel zinc finger gene family and likely functions as a transcription factor. It possesses a nuclear localization signal (NLS) similar to the bipartite basic NLS of nucleoplasmin, but unlike nucleoplasmin, which re-enters nuclei immediately after fertilization, xnf7 remains cytoplasmic until the mid-blastula transition (MBT). We have measured the accumulation of injected labeled xnf7 protein or protein produced from synthetic xnf7 transcripts in the oocyte nuclei (GV). The data show that the NLS of xnf7 functions efficiently in oocytes. Mutations in either of the bipartite basic domains of the xnf7 NLS inhibit nuclear accumulation, while mutations in the spacer sequences have no effect. The xnf7 NLS linked to pyruvate kinase directs the efficient accumulation of this protein into nuclei of early embryos prior to the MBT. These data suggest that retention of the xnf7 protein during development is the result of a mechanism that interferes with the xnf7 NLS function. PMID- 8408274 TI - Monolayer and aggregate cultures of rainbow trout hepatocytes: long-term and stable liver-specific expression in aggregates. AB - An aggregate culture system for rainbow trout hepatocytes was developed to study liver-specific mRNA expression. Maintenance of differentiated functions and morphology of hepatocytes were examined using both monolayer and aggregate systems. The rainbow trout estrogen receptor and vitellogenin genes were induced by estradiol and their mRNAs used as markers of cell differentiation during cell culture. In monolayer culture, rainbow trout hepatocytes established very few cell-cell contacts in minimal media. The use of more complete media promotes cell cell contacts and cell islet formation. Hepatocyte response to estradiol stimulation was generally lower than in vivo but a correlation between the degree of cellular organization and the intensity of the hormonal response was observed. However, in this system hepatocytes progressively lost their specific hormonal response between 5 and 10 days. In aggregates with DMEM/F12 and Ultroser SF, cell cell contacts were maximized and stabilized during at least one month. The levels of rainbow trout estrogen receptor and vitellogenin mRNAs induced by estradiol were stable and maintained at a level comparable to in vivo levels; vitellogenin synthesis and secretion remained fully functional for the duration of the culture. PMID- 8408275 TI - Alveolar type II cell-fibroblast interactions, synthesis and secretion of surfactant and type I collagen. AB - During alveolar development and alveolar repair close contacts are established between fibroblasts and lung epithelial cells through gaps in the basement membrane. Using co-culture systems we have investigated whether these close contacts influence synthesis and secretion of the principal surfactant apoprotein (SP-A) by cultured rat lung alveolar type II cells and the synthesis and secretion of type I collagen by fibroblasts. The alveolar type II cells remained cuboidal and grew in colonies on fibroblast feeder layers and on Matrigel-coated cell culture inserts but were progressively more flattened on fixed fibroblast monolayers and plastic. Alveolar type II cells cultured on plastic released almost all their SP-A into the medium by 4 days. Alveolar type II cells cultured on viable fibroblasts or Matrigel-coated inserts above fibroblasts accumulated SP A in the medium at a constant rate for the first 4 days, and probably recycle SP A by endocytosis. The amount of mRNA for SP-A was very low after 4 days of culture of alveolar type II cells on plastic, Matrigel-coated inserts or fixed fibroblast monolayers: relatively, the amount of mRNA for SP-A was increased 4 fold after culture of alveolar type II cells on viable fibroblasts. Co-culture of alveolar type II cells with confluent human dermal fibroblasts stimulated by 2- to 3-fold the secretion of collagen type I into the culture medium, even after the fibroblasts' growth had been arrested with mitomycin C. Collagen secretion, by fibroblasts, also was stimulated 2-fold by conditioned medium from alveolar type II cells cultured on Matrigel. The amount of mRNA for type I collagen increased only modestly when fibroblasts were cultured in this conditioned medium. This stimulation of type I collagen secretion diminished as the conditioned medium was diluted out, but at high dilutions further stimulation occurred, indicating that a factor that inhibited collagen secretion also was being diluted out. The conditioned medium contained low levels of IGF-1 and the stimulation of type I collagen secretion was abolished when the conditioned medium was pre-incubated with antibodies to insulin-like growth factor 1 (IGF-1). There are important reciprocal interactions between alveolar type II cells and fibroblasts in co-culture. Direct contacts between alveolar type II cells and fibroblasts appear to have a trophic effect on cultured alveolar type II cells, increasing the levels of mRNA for SP-A. Rat lung alveolar type II cells appear to release a factor (possibly IGF-1) that stimulates type I collagen secretion by fibroblasts. PMID- 8408276 TI - An in vitro model for the analysis of intestinal brush border assembly. I. Ultrastructural analysis of cell contact-induced brush border assembly in Caco 2BBe cells. AB - Intestinal epithelial cells assemble and maintain a polarized, highly organized membrane-cytoskeleton array, the brush border. We describe an in vitro, cell contact-induced brush border assembly model using the Caco-2BBe clones. Subconfluent cells were 'depolarized' by brief passage through suspension culture in the presence of cytochalasin D and re-plated on filters at high density in low Ca2+ medium. Upon return to regular medium, these small, rounded cells with bleb like protrusions formed, over the course of 19 days, a polarized monolayer of tall, columnar cells with a well-defined brush border. Ultrastructural changes were documented by both transmission and scanning electron microscopy. The earliest events of microvillar assembly coincided with a short period of cell aggregation. Intercellular cysts were occasionally observed within these aggregates, and junction formation between cells which had no contact with the filter was also observed. Monolayer formation was completed within 48 hours, and cell height steadily increased approximately 3.5-fold over 19 days. Concurrent with monolayer formation and the increase in cell height, sparse microvilli with a few actin core filaments gradually became more dense and better organized. By the third day, the actin core bundles had begun to extend into the subjacent cytoplasm, while terminal web assembly was underway at five days. The mature morphology of the brush border was first observed at nine days, although cell height and microvillar density continued to increase during the subsequent ten days. Microvillar density rose approximately nine-fold throughout brush border assembly in the Caco-2BBe cells. With the exception of the formation of cellular aggregates at the onset of the time course, this sequence of morphological changes is comparable to that observed during brush border assembly in embryonic intestinal epithelial cells. The Caco-2BBe assembly model provides a useful system in which to investigate various molecular aspects of brush border assembly. PMID- 8408277 TI - An in vitro model for the analysis of intestinal brush border assembly. II. Changes in expression and localization of brush border proteins during cell contact-induced brush border assembly in Caco-2BBe cells. AB - In the companion paper (M. D. Peterson and M. S. Mooseker (1993). J. Cell Sci. 105, 445-460) we describe a method for modeling brush border assembly in the Caco 2BBe clones. In this study we have examined the molecular changes accompanying cell contact-induced brush border assembly. A subset of brush border proteins was tracked throughout brush border assembly by immunoblotting and by immunofluorescent localization using laser scanning confocal microscopy. Actin, fodrin, villin and presumptive unconventional myosin immunogens were distributed at the periphery of depolarized cells. All proteins partitioned primarily with the membrane fraction upon differential sedimentation of depolarized cell lysates; the fractionation patterns were comparable to those of confluent cells. After a monolayer had formed, each protein showed a redistribution to the apical domain in a discrete sequence. Actin and villin began to shift apically at 2 d, while fodrin and the unconventional myosin immunogens did not redistribute until 3 d. Enterocyte-like localization was observed by 5 d for all proteins. Sucrase isomaltase was not reliably detectable until 9 d by immunofluorescence, after brush border assembly was complete. Quantitative immunoblot analysis of total cell extracts demonstrated an average 10-fold increase in villin levels, while fodrin levels appeared to remain unchanged. Three putative unconventional myosin immunogens of 140 kDa, 130 kDa, and 110 kDa have been detected previously in the C2BBe cells with a head-specific monoclonal antibody to avian brush border myosin I (M. D. Peterson and M. S. Mooseker (1992) J. Cell Sci. 102, 581-600). Each of these immunogens displayed distinct expression patterns during brush border assembly. The 140 kDa species decreased by half, while the 130 kDa immunogen(s) did not change in any consistent fashion. The 110 kDa protein, presumed to be human brush border myosin I, rose on average 8-fold. A ribonuclease protection assay was also performed using a probe for human brush border myosin I. Equal amounts of total RNA from depolarized and confluent cells were assayed; the level of protected product was approximately 9-fold greater in the confluent cells. The expression patterns of the brush border proteins, coupled with the correlation to the ultrastructural features during brush border assembly in C2BBe cells, show that differentiation of the C2BBe cells closely resembles the changes that occur during human fetal intestinal differentiation. PMID- 8408278 TI - Inhibition of protein synthesis separates autophagic sequestration from the delivery of lysosomal enzymes. AB - To investigate the role of newly synthesized proteins during autophagic sequestration and degradation, the effects of protein synthesis inhibition on autophagic vacuole (AV) formation and degradation were analyzed. The inhibition of protein synthesis was found to separate autophagic sequestration from the delivery of lysosomal enzymes to (AVs). Pretreatment with cycloheximide for > or = 3 h caused a drastic inhibition of autophagy-induced degradation. Surprisingly, morphological analyses showed that the inhibition of protein synthesis for up to 12 h did not block the formation of nascent AVs; however, it did prevent their conversion into degradative AVs. Using immunoperoxidase cytochemistry with an antibody against cathepsin D and labeling of lysosomes with endocytosed colloidal gold, we found that the nascent AVs that formed during prolonged cycloheximide pretreatment had not received lysosomal markers. The inhibition of autophagic degradation and lysosomal enzyme delivery were rapidly reversed following the removal of cycloheximide. These results suggest that there is a fairly rapid turnover of protein(s) that are necessary for lysosomal fusion, but that the initial formation of AVs is independent of new protein synthesis for a long period of time. PMID- 8408279 TI - Use of a general purpose mammalian expression vector for studying intracellular protein targeting: identification of critical residues in the nuclear lamin A/C nuclear localization signal. AB - We have constructed a general purpose mammalian expression vector for the study of intracellular protein targeting. The vector, p3PK, facilitates construction of N- and/or C-terminal fusions of an amino acid sequence of interest to the normally cytosolic protein chicken muscle pyruvate kinase (CMPK). The vector has been engineered such that any fusion construct can be subcloned into the versatile pJx omega family of mammalian expression vectors and into pGEX bacterial expression vectors, for the generation of affinity reagents. In this paper, we demonstrate the general utility of p3PK by redirecting CMPK to mitochondria (using the twelve amino acid pre-sequence of yeast cytochrome c oxidase subunit IV) and to the nucleus (using a putative eight amino acid nuclear localization signal from human nuclear lamins A and C). We also report that, contrary to the predictions of previously published work, substitution of a critical residue in the nuclear lamin A/C nuclear localization signal (the equivalent of lysine 128 in the SV40 large T nuclear localization signal) retains nuclear localization, and discuss how amino acid context might affect targeting to the nucleus. PMID- 8408280 TI - Controlled induction of focal adhesion disassembly and migration in primary fibroblasts. AB - Fibroblast migration is an integral component of biological processes such as wound healing and embryogenesis. Previous experiments examining fibroblast locomotion from tissue explants have shown that migrating fibroblasts lack, or contain only transient, focal adhesions (focal contacts). Focal adhesions are specialized regions of tight cell-matrix interaction, assembled by a complex process of transmembrane signalling. Although the explant model has been used for studying several aspects of fibroblast locomotion, it is limited by the lack of control over migration, and only a small percentage of the cells actually locomoting. Therefore, we have developed an in vitro model for cultured fibroblast strains where the presence or absence of focal adhesions can be manipulated, and in the latter case 70% of these cells become locomotory. The stimulus used to decrease the percentage of cells containing focal adhesions, and hence enhance locomotion, was newborn rat heart-conditioned medium (HCM). Addition of HCM to rat embryo fibroblasts induced both chemokinesis and chemotaxis. Cells disassembled focal adhesions on a variety of extracellular matrix substrates after approximately 6 h of stimulation with HCM; conversely, removal of HCM promoted reformation of focal adhesions within 12-24 h. HCM stimulated fibroblasts which lacked focal adhesions concomitantly lacked F-actin stress fibers and focal concentrations of vinculin and talin. Therefore, fibroblast migration can be readily controlled in an on-off manner through conditioned medium, which influences the absence or presence of focal adhesions. PMID- 8408281 TI - CO2 production in cell-free extracts of fission yeast detects cell cycle changes. AB - CO2 production was followed by manometry in starved cell-free extracts of fission yeast stimulated by unstarved cell-free extracts from a synchronous culture. The degree of stimulus, measured by the lag time in CO2 production, varied markedly during the cell cycle, with a maximum for cells at about mitosis and a minimum for septated cells. Similar differences in lag time were found with unstarved extracts of cdc13.117 grown at 37 degrees C and 35 degrees C. PMID- 8408282 TI - Involvement of annexin II in DNA replication: evidence from cell-free extracts of Xenopus eggs. AB - Cell-free extracts of Xenopus eggs efficiently initiate and complete semiconservative DNA replication of exogenously added plasmid DNA. DNA replication in such extracts can be neutralized by a monoclonal antibody (D1/274.5) against human annexin II. Specific immunodepletion of Xenopus annexin II from the egg extracts results in loss of DNA replicative ability. Immunodepletion of annexin II does not prevent nuclear assembly, a stringent requirement for DNA synthesis on exogenous DNA in this system. Replicative ability can be restored to the immunodepleted extracts by the addition of purified human annexin II. These results demonstrate that annexin II is involved in chromosomal DNA replication and has a role in the cell cycle of higher eukaryotes. PMID- 8408283 TI - Visualization of replication sites in unfixed human cells. AB - Sites of DNA replication in nuclei are focally concentrated, suggesting that an underlying structure organizes the activity of many polymerases. As fixation could induce aggregation into foci, we examined the distribution of replication sites in unfixed nuclei. HeLa cells were encapsulated in agarose microbeads, permeabilized in a 'physiological' buffer, their DNA polymerizing activity characterized, and replication sites directly labelled by incubation with fluorochrome-dUTP conjugates. Using conventional and digital fluorescence microscopy, 80-250 foci were seen in these unfixed cells. These foci are unlikely to be formed by the aggregation of separate polymerases as most replication activity found in vivo is retained throughout these procedures. Although commonly used fixation methods collapsed or dispersed their periphery, the central core was very stable. Foci remained when approximately 90% chromatin was removed, suggesting they were attached to an underlying structure. PMID- 8408284 TI - Transitions between in situ and isolated chromatin. AB - We show that the mechanism by which chromatin displaying higher-order structure is usually isolated from nuclei involves a transition to an extended nucleosomal arrangement. After being released from nuclei, chromatin must refold in order to produce the typical chromatin fibers observed in solution. For starfish sperm chromatin with a long nucleosome repeat (222 bp), isolated fibers are significantly wider than those in the nucleus, indicating that the refolding process does not regenerate the native higher-order structure. We also propose that for typical eukaryotic nuclei, the concept that the native state of the (inactive) bulk of the genome is a chromatin fiber with defined architecture be reconsidered. PMID- 8408285 TI - Topoisomerase II inhibition prevents anaphase chromatid segregation in mammalian cells independently of the generation of DNA strand breaks. AB - Yeast temperature-sensitive mutants of DNA topoisomerase II are incapable of chromosome condensation and anaphase chromatid segregation. In mammalian cells, topoisomerase II inhibitors such as etoposide (VP-16-123) have similar effects. Unfortunately, conclusions drawn from work with mammalian cells have been limited by the fact that the standard inhibitors of topoisomerase II also generate DNA strand breaks, which when produced by other agents (e.g. ionizing radiation) are known to affect progression into and through mitosis. Here we show that the anti tumour agent ICRF-193, recently identified as a topoisomerase II inhibitor operating by a non-standard mechanism, generates neither covalent complexes between topoisomerase II and DNA, nor adjacent DNA strand breaks, in mitotic HeLa. However, the drug does prevent anaphase segregation in HeLa and PtK2 cells, with effects similar to those of etoposide. We therefore conclude that topoisomerase II function is required for anaphase chromosome segregation in mammalian cells, as it is in yeast. PMID- 8408286 TI - Ultraviolet microbeam irradiation of chromosomal spindle fibres in Haemanthus katherinae endosperm. I. Behaviour of the irradiated region. AB - We used an ultraviolet microbeam to irradiate chromosomal spindle fibres in metaphase Haemanthus endosperm cells. An area of reduced birefringence (ARB) was formed at the position of the focussed ultraviolet light with all wavelengths we used (260, 270, 280, and 290 nm). The chromosomal spindle fibre regions (kinetochore microtubules) poleward from the ARBs were unstable: they shortened (from the ARB to the pole) either too fast for us to measure or at rates of about 40 microns per minute. The chromosomal spindle fibre regions (kinetochore microtubules) kinetochore-ward from the ARBs were stable: they did not change length for about 80 seconds, and then they increased in length at rates of about 0.7 microns per minute. The lengthening chromosomal spindle fibres sometimes grew in a direction different from that of the original chromosomal spindle fibre. The chromosome associated with the irradiated spindle fibre sometimes moved off the equator a few micrometers, towards the non-irradiated half-spindle. We discuss our results in relation to other results in the literature and conclude that kinetochores and poles influence the behaviour of kinetochore microtubules. PMID- 8408287 TI - Regulation of the intracellular distribution of cytoplasmic dynein by serum factors and calcium. AB - Previous work has indicated that cytoplasmic dynein localizes primarily to lysosomes in cultured fibroblasts, consistent with a function for dynein in retrograde movement. We now show that dynein can be redistributed from a lysosome associated pool to a more diffuse cytoplasmic pool upon shifting fibroblasts to culture medium lacking serum for several hours. This effect on dynein localization is readily reversed upon addition of serum, with a substantial return to a control appearance of punctate staining within 10 minutes. The serum effect appears to be selective for dynein, in that the localization of kinesin and the overall morphology of intracellular organelles does not change. However, the distribution of kinesin-positive vesicles and lysosomes does appear to be altered during serum starvation, in that these organelles are located to greater extents in the peripheral regions of the cell. Dynein is also associated with the mitotic apparatus, but this localization does not change in response to serum starvation. Removal of calcium from the extracellular medium also results in the loss of punctate dynein staining, which can be recovered upon addition of calcium to calcium-free medium. The redistribution of dynein observed under these experimental conditions may reflect the activity of a regulatory process controlling the association of dynein with organelles, thereby providing one means of modulating intracellular transport. PMID- 8408288 TI - Primary structure and microtubule-interacting domain of the SP-H antigen: a mitotic MAP located at the spindle pole and characterized as a homologous protein to NuMA. AB - Using a human autoantibody, SP-H, we identified a 200-230 kDa mitotic MAP in a variety of mammalian cell lines which shows affinity for the minus end of microtubules and also becomes associated with the spindle pole during mitosis. To examine the detailed structure and functional organization of the protein, the gene coding for the end-specific MAP was isolated and characterized by screening a human placenta lambda gt11 expression library using SP-H as a probe. Overlapping cDNA clones, which covered the entire length of the coding region of the SP-H antigen, were obtained. Polyclonal antibodies raised against fusion proteins generated from non-overlapping cDNA fragments stained the HeLa SP-H antigen in interphase and mitotic cells, and recognized a single 215 kDa band on immunoblots, as did the original SP-H antibody. Analysis of the nucleotide sequence revealed a 7,091 nucleotide sequence with an open reading frame of 6,345 nucleotides encoding a 2,115 amino acid polypeptide with a calculated molecular mass of 238,376 Da. The predicted amino acid sequence showed the protein to be composed of an alpha-helical domain, flanked by globular domains located at the amino and carboxy termini. The sequence contained five repeats of the hypothetical leucine zipper motif: one is in the N-terminal globular domain, and four are in the central alpha-helical stalk. Comparison with other sequences in the database shows that the SP-H antigen is identical to the NuMA protein reported by Yang et al. (1992) J. Cell Biol. 116, 1303-1317, but there are differences between the SP-H antigen and NuMA sequence reported by Compton et al. (1992) J. Cell Biol. 116, 1395-1408. cDNA inserts of the truncated SP-H antigen were expressed in both insect Sf9 cells and in cultured mammalian cells. The recombinant protein corresponding to the C-terminal half of the protein was restricted to the nucleus, whereas the N-terminal half of the protein was localized in the cytoplasm, suggesting the presence of a nuclear translocation signal(s) in the C-terminal domain. The C-terminal polypeptide expressed in mitotic COS cells was shown to specifically localize at the spindle pole. Microtubule-binding assays using in vitro transcribed/translated polypeptide products from different domains of the SP-H antigen further suggested that the SP H antigen interacts with microtubules through the globular domain at the C terminus. PMID- 8408289 TI - Heterogeneity of thymic stromal cells and thymocyte differentiation: a cell culture approach. PMID- 8408290 TI - Regulation of p53 protein expression in human breast cancer cell lines. AB - Mutation of the p53 gene is a common occurrence in human breast cancers but is by no means universal. However, even in tumours where the gene is not mutated altered levels of p53 protein are often detected. This is also observed in cell lines derived from human breast cancers. By transfecting such cell lines containing either wild type or mutant p53 genes with a temperature-sensitive mutant mouse p53 gene we have established that the cellular environment plays a critical role in the regulation of p53 protein expression. The results suggest that tumours that aberrantly express wild-type p53 may have lost the normal growth regulatory response to the protein and thus be functionally similar to those expressing the mutant protein. PMID- 8408291 TI - Tyrosine phosphorylation of the focal adhesion kinase pp125FAK during development: relation to paxillin. AB - Significant changes in the level of protein tyrosine phosphorylation accompany avian embryonic development. A comparison of different tissues reveals that a similar and remarkably restricted complement of proteins is modified in this manner. In each case the major proteins detected using anti-phosphotyrosine antibodies have molecular masses of approximately 170, 150, 125, 70 and 50 kDa. As a first step in determining the function of this protein modification in embryogenesis we have initiated a study to identify these phosphoproteins. We have previously reported that the 70 kDa band is paxillin, a component of actin membrane attachment sites associated with regions of cell adhesion (Turner, C. E. (1991) J. Cell Biol. 115, 201-207). We report here that the 125 kDa phosphotyrosine-containing protein is the tyrosine kinase pp125FAK, a protein that co-localizes with paxillin at sites of adhesion (Schaller et al. (1992) Proc. Nat. Acad. Sci. USA 89, 5192-5196). Tyrosine phosphorylation of both pp125FAK and paxillin was detected at low levels as early as embryonic day 3 and increased steadily during the first half of development, reached a maximum between embryonic days eight and twelve, and declined to background levels prior to hatching. Paxillin protein expression also increased during the first half of embryogenesis, suggesting little change in the overall phosphorylation of this protein through embryonic day 8. In contrast, pp125FAK, following an initial increase, is expressed at a constant high level during these early embryonic stages, implying an increase in its overall phosphotyrosine content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408292 TI - Binding of the alpha-fodrin SH3 domain to the leading lamellae of locomoting chicken fibroblasts. AB - Fodrin (nonerythroid spectrin) is a membrane skeletal protein that plays an important role in the establishment and maintenance of the cell shape and polarity. We have identified in alpha-fodrin an src homology 3 (SH3)-related region, a small domain that is present in a large number of proteins that are involved in signal transduction, cell polarization and membrane-cytoskeleton interactions. In this study we have explored the function of the alpha-fodrin SH3 by incubating fixed and permeabilized cultured chicken fibroblasts with the alpha fodrin SH3 peptide, expressed in bacteria as a fusion protein with glutathione S transferase. Immunofluorescence and immunoelectron microscopy showed that alpha fodrin SH3 binds to the cytoplasmic face of the plasma membrane in the leading lamellae and the pseudopodial lobes of the spreading and locomoting cells. No, or only minimal, binding was seen in immotile cells, or in the stationary trailing ends of the locomoting cells. SH3 binding was also seen in cytochalasin-D-treated cells, suggesting that actin filaments are not responsible for the binding. These findings suggest that alpha-fodrin SH3 interacts with plasma membrane components that are present in the leading lamellae exclusively or are modulated in a manner specific to the leading lamellae. PMID- 8408293 TI - Aggregation of band 3 in hereditary ovalocytic red blood cell membranes. Electron microscopy and protein rotational diffusion studies. AB - Microaggregation of band 3 proteins in hereditary ovalocytic membranes was investigated by rotational diffusion measurements and by electron microscopy. It was previously shown that band 3 in ovalocytic membranes has decreased rotational mobility compared with band 3 in normal cells (Tilley, L., Nash, G.B., Jones, G.L. and Sawyer, W.L. (1991) J. Membr. Biol. 121, 59-66). This result could arise from either altered interactions with cytoskeletal proteins or from band 3 microaggregation. In the present study it was found that removal of spectrin and actin from the membrane had no effect on the rotational mobility of ovalocytic band 3. Additional removal of ankyrin and band 4.1, as well as cleavage of the cytoplasmic domain of band 3 with trypsin, did enhance band 3 mobility, as is the case in the membranes from normal cells. However, the rotational mobility of ovalocytic band 3 was always considerably less than that of normal band 3 under the same conditions. Scanning electron microscopy and low power electron micrographs of freeze-fracture replicas revealed that the surfaces of ovalocytes were more irregular than those of normal erythrocytes. At higher magnification, numerous linearly arranged intramembranous particles were observed on the P-faces of freeze-fractured ovalocytes but not on normal cells. These clusters consist of straight or slightly curved lines of 10-15 particles in single rows. From these results it is deduced that the reduced rotational mobility of band 3 in ovalocytes is a consequence of the formation of microaggregates, which are very probably induced by the mutation in the membrane-bound domain of ovalocytic band 3. PMID- 8408294 TI - Avian pluripotent haemopoietic progenitor cells: detection and enrichment from the para-aortic region of the early embryo. AB - In the avian embryo, the wall of the aorta is a site where haemopoiesis occurs in large diffuse foci from day 3 to day 10. In contrast to haemopoiesis in other organs of the embryo, para-aortic haemopoiesis is sustained by stem cells, which emerge in situ. Previous studies have demonstrated that the para-aortic region from the day-4 chick embryo harbours committed myeloid progenitors and committed erythroid progenitors. The present paper reports the in vitro development of para aortic progenitors with both myelomonocytic and erythroid potentialities. Three types of myelo-erythroid progenitors were observed, giving rise to erythroblasts and monocytes, to erythroblasts and granulocytes, or to erythroblasts, monocytes and granulocytes. Their frequency in the para-aortic cell suspension was 1 per 10,000 cells. In cell sorting experiments, they co-sorted with committed progenitors in the cell population that immunolabeled with the VI-A2 monoclonal antibody, which is specific for chicken haemopoietic cells. Cell sorting also demonstrated that these multipotential progenitors did not express the BEN cell surface molecule, in contrast to late myeloid progenitors. The BEN molecule belongs to the immunoglobulin superfamily and is expressed by haemopoietic progenitors from bone marrow, selective sets of neurons and epithelial cells from the bursa of Fabricius. The myelo-erythroid progenitors were enriched 4 times in the VI-A2-positive cell population, and 2 to 5 times in the BEN-negative population. These results represent the first in vitro demonstration of avian normal myelo-erythroid progneitors. PMID- 8408295 TI - F-actin content and spatial distribution in resting and chemoattractant stimulated human polymorphonuclear leucocytes. Which role for intracellular free calcium? AB - Intracellular free calcium concentration ([Ca2+]i) plays a pivotal role for many responses in polymorphonuclear leucocytes (PMNs) stimulated by chemoattractants such as N-formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe). The importance of [Ca2+]i in the morphological polarization was investigated by using calcium manipulated PMNs. We loaded human PMNs with BAPTA/AM to buffer or chelate [Ca2+]i in the presence or the absence of extracellular calcium by using fluo-3/AM as calcium indicator. The shape changes of PMNs were determined by microscopic examination, and membrane ruffling by right-angle light-scatter changes. Actin polymerization and F-actin distribution were recorded by staining PMNs with bodipy-phallacidin and quantified by quantitative fluorescence microscopy. We found that calcium-free incubation of PMNs loaded or not with 50 microM BAPTA/AM did not modify morphological polarization, membrane ruffling, actin assembly and F-actin distribution of PMNs stimulated with fMet-Leu-Phe, suggesting that these responses were probably functionally linked. It should be noted that incubation of PMNs in calcium-free conditions resulted in a radial distribution of F-actin and a moderate polymerization of actin, but not in morphological polarization of PMNs. Moreover, both calcium-sensitive and calcium-insensitive mechanisms of actin polymerization were additive, and inhibitable by 5 micrograms/ml cytochalasin B. PMID- 8408296 TI - Interaction of the human autoantigen p150 with splicing snRNPs. AB - An important goal of studies on pre-mRNA splicing is to identify factors that mediate the snRNP-snRNP and snRNP-pre-mRNA interactions that take place in the spliceosome. The U4/U6 snRNP is one of the four snRNPs that are subunits of spliceosomes. A rare patient autoimmune serum (MaS serum) has recently been identified that specifically immunoprecipitates U4/U6 snRNP from HeLa cell extracts through recognition of a 150 kDa autoantigen (p150) (Okano and Medsger, Journal of Immunology, 146, 535-542, 1991). Here we show that in addition to U4/U6 snRNP, p150 can also be detected associated with 20 S U5, U4/U6.U5 and 17 S U2 snRNPs, but not with U1 snRNP. In each particle p150 is present in sub stoichiometric levels relative to the major snRNP proteins. We show that MaS serum selectively immunoprecipitates a sub-population of U4/U6 snRNPs in which the m3G-cap structure is masked and that p150 is preferentially associated with U6 snRNA in the U4/U6 particle. Anti-p150 antibodies show widespread nucleoplasmic staining, excluding nucleoli, with an elevated concentration in coiled bodies. This changes to a discrete punctate pattern when cells are treated with alpha-amanitin. Both the cytological and biochemical data indicate that the p150 autoantigen is a snRNP-associated factor in vivo. We also present biochemical evidence confirming that assembly of U4/U6 and U5 snRNPs into a U4/U6.U5 tri-snRNP particle is an integral step in the spliceosome assembly pathway. Addition of the purified U4/U6.U5 tri-snRNP restores splicing activity to inactivated HeLa nuclear extracts in which splicing had been inhibited by specific depletion of either the U4/U6 or U5 snRNPs. PMID- 8408297 TI - Polarized distribution of Listeria monocytogenes surface protein ActA at the site of directional actin assembly. AB - The facultative intracellular pathogen Listeria monocytogenes can infect host tissues by using directional actin assembly to propel itself from one cell into another. The movement is generated by continuous actin assembly from one end of the bacterium into a tail, which is left behind in the cytoplasm. Bacterial actin assembly requires expression of the bacterial gene actA. We have used immunocytochemistry to show that the actA gene product, ActA, is distributed asymmetrically on the bacterial surface: it is not expressed at one pole and is increasingly concentrated towards the other. This polarized distribution of ActA was linked to bacterial division: ActA protein was not, or only faintly, expressed at the pole that had been formed during the previous division. On intracellular bacteria ActA was expressed at the site of actin assembly, suggesting that ActA may be involved in actin filament nucleation off the bacterial surface. We predict that the asymmetrical distribution of this protein is required for the ability of intracellular Listeria to move in the direction of the non-ActA expressing pole. PMID- 8408298 TI - Heat shock results in cell cycle delay and synchronisation of mitotic domains in cellularised Drosophila melanogaster embryos. AB - Cells of Drosophila embryos that are subjected to a 37 degrees C temperature shock whilst undergoing the S-phase of cell cycle 14 arrest with their microtubules in an interphase-like state, and with nuclei showing unusual chromatin condensation. They do not recover from this state within a 30 minute period even though extensive gastrulation movements can occur. Cells of embryos heat shocked in G2-phase are delayed in interphase with high levels of cyclins A and B. Within ten minutes recovery from heat shock, cells enter mitosis throughout the embryo. The degradation of the mitotic cyclins A and B in these synchronised mitotic domains does not follow the normal timing, but is delayed. These findings point to a need for caution when interpreting experiments that use the heat shock promoter to study the expression of cell cycle control genes in Drosophila. PMID- 8408299 TI - Redundant enhancer elements guide beta 1 tubulin gene expression in apodemes during Drosophila embryogenesis. AB - During Drosophila embryogenesis, the beta 1 tubulin gene (beta Tub56D) is expressed in the CNS and PNS as well as in the apodemes. In this report we determine the regulation of beta 1 tubulin gene expression during formation of the attachment sites of the somatic muscles, the apodemes. The process was analysed in transgenic flies carrying beta 1 enhancer-hsp70 promoter-lacZ fusion constructs. Expression is first detected at late stage 13 and remains until hatching. By deletion analysis of the intron we identified a 14-bp element present in three copies. This element represents a classical enhancer, as it acts on a heterologous promoter. Separate fragments containing the respective elements yield nearly identical expression patterns, and no cooperativity was observed between the three copies. Thus, the expression of the beta 1 tubulin gene in the apodemes is under control of redundant enhancer elements. Double staining for beta 1 tubulin gene expression in apodemes and for beta 3 tubulin gene expression in muscles allowed us to correlate apodeme and muscle formation. Cells of the apodemes that are in contact with their corresponding muscles show expression of the reporter gene as monitored by antibody staining. PMID- 8408300 TI - Expression of high molecular weight tau in the central and peripheral nervous systems. AB - Using a novel PCR approach, we have cloned a cDNA encoding the entire high molecular weight tau molecule from rat dorsal root ganglia. The resulting 2080 bp cDNA differs from low molecular weight rat brain tau by the insertion of a novel 762 bp region (exon 4a) between exons 4 and 5. This cDNA clone is identical in sequence with a high molecular weight tau (HMW) cDNA from rat PC12 tumor cells and is closely related to a HMW tau cDNA from mouse N115 tumor cells. In vitro transcription/translation produces a protein that migrates on SDS-PAGE with the same apparent molecular weight as HMW tau purified from rat sciatic nerve. The HMW tau protein is generated from an 8 kb mRNA, which can be detected by northern blots in peripheral ganglia, but not in brain. A more sensitive assay using PCR and Southern blot analysis demonstrates the presence of exon 4a in spinal cord and in retina. In combination with immunohistochemical studies of spinal cord, these data suggest that HMW tau, though primarily in the peripheral nervous system, is also expressed in limited areas of the central nervous system, although its presence cannot be detected in the cerebral cortices. PMID- 8408301 TI - Alpha 3 beta 1 integrin is moved into focal contacts in kidney mesangial cells. AB - The movement of integrins into focal adhesive structures accompanies cell attachment to extracellular matrix. The kinetics of incorporation of integrins into focal contacts was studied during attachment to matrix of mesangial cells of the kidney glomerulus. On collagen, fibronectin, laminin and vitronectin, the number and intensity of talin-focal contacts increased with time. Talin containing focal contacts were present in mesangial cells within 2 h of plating and in control cells (HT1080 and Rugli) within 1 h. Integrin alpha-chains colocalized with talin, dependent on the matrix substrate. The attachment, spreading and organization of integrin into focal contacts was not affected when endogenous protein synthesis was suppressed with cycloheximide. In Rugli, alpha 1 beta 1 organized into focal contacts on collagen and laminin, while in HT1080 alpha 2 beta 1 organized on collagen type I, alpha 5 beta 1 on fibronectin, alpha 6 beta 1 on laminin, and alpha 3 beta 1 and alpha 4 beta 1 were diffusely distributed on all substrates. These distributions mirrored the usage and expression patterns previously established for integrins in these cells and was as predicted from the literature. In mesangial cells, however, alpha 3 beta 1 was also organized into prominent focal contact arrays on collagen, fibronectin, EHS and human placental laminins, but not on vitronectin, while alpha 6 beta 1 was not organized. Initial attachment and spreading of mesangial cells was absolutely dependent on divalent cations. Mg2+ and Mn2+ supported attachment on all substrates, while Ca2+ stimulated attachment on laminin (E8), fibronectin and vitronectin. The data suggest that the functional integrins on mesangial cells include alpha 1 beta 1 (on collagen and laminin) alpha 2 beta 1 (on collagen), alpha 5 beta 1 (on fibronectin) and alpha V beta 3 (on vitronectin). However, mesangial cells do not use alpha 6 beta 1 on laminin, and the data support a role for alpha 3 beta 1 as putative receptor for fibronectin, collagen and laminin. PMID- 8408302 TI - The matrix secreted by 804G cells contains laminin-related components that participate in hemidesmosome assembly in vitro. AB - Hemidesmosomes are important adhesion devices found in epithelial cells. They connect the intermediate filament cytoskeleton network with components of the basement membrane zone. 804G cells are an unusual epithelial cell line, since they form bona fide hemidesmosomes when plated on glass or plastic. In this study we tested an hypothesis: that this ability is a consequence of an extracellular component produced by the 804G cells. As probes for our study we generated a rabbit antiserum (J18) and monoclonal antibodies against components of urea solubilized 804G matrix. Antibodies in the J18 serum recognize major lectin binding polypeptides of 150, 140 and 135 kDa in the 804G matrix. A monoclonal antibody (5C5) that shows reactivity with the 150 and 135 kDa polypeptides in western immunoblots immunoprecipitates all three molecular mass species, indicating that these polypeptides are part of a matrix complex. Moreover, one, at least, of these matrix elements is immunologically related to laminin, since J18 antibodies selected on fusion protein fragments of a newly characterized laminin variant, laminin B2t (Kallunki et al., J. Cell Biol., 119, 679-694, 1992), react with the 140 kDa polypeptide component of the 804G cell matrix. To undertake functional analyses of 804G matrix, cells of the human epidermal carcinoma line SCC12, which do not assemble bona fide hemidesmosomes in vitro, were cultured on 804G matrix for 24 h and then analysed by confocal immunofluorescence and electron microscopy. In SCC12 cells maintained on 804G cell matrix, hemidesmosomal antigens localize in a distinctive leopard spot pattern that mirrors the distribution of 804G matrix elements. Furthermore, ultrastructural analysis reveals that the 804G cell matrix supports the formation of 'mature' hemidesmosomes by SCC12 cells. Thus 804G cell matrix is a remarkable tool for hemidesmosome studies and it will now be of great importance to determine the exact composition of the 804G matrix, especially its structural and antigenic relationship to laminins. PMID- 8408303 TI - Villin-induced growth of microvilli is reversibly inhibited by cytochalasin D. AB - Villin is an actin-binding protein that is associated with the cytoskeleton of brush border microvilli. In vitro, villin nucleates, caps or severs actin filaments in a Ca(2+)-dependent manner. In the absence of Ca2+, villin organizes microfilaments into bundles. Transfection of a villin-specific cDNA into cultured cells that do not produce this protein results in the growth of long surface microvilli and the reorganization of the underlying actin cytoskeleton. Here we studied the effects of low concentrations of cytochalasin D on the induction of these plasma membrane-actin cytoskeleton specializations. Transfected cells were treated with concentrations of cytochalasin D that prevent the association of actin monomers with the fast-growing end of microfilaments in vitro. In villin positive cells, cytochalasin D inhibited the growth of microvilli and promoted the formation of rodlet-like actin structures, which were randomly distributed throughout the cytoplasm. The formation of these structures was dependent on large amounts of villin and on the integrity of an actin-binding site located at the carboxy terminus of villin, which is required for microfilament bundling in vitro and for the growth of microvilli in vivo. The effect of cytochalasin D was reversible. The observation of living cells by video-imaging revealed that when cytochalasin D was removed, rapid disassembly of actin rodlets occurred after a lag phase. The present data stress the important role of the plasma membrane in the organization of the actin cytoskeleton and suggest that the extension of the microvillar plasma membrane is dependent on the elongation of microfilaments at their fast-growing end. Inhibition of microfilament elongation near the plasma membrane by cytochalasin D may result in the 'random' nucleation of actin filaments throughout the cytoplasm. On the basis of the present data, we propose that villin is involved in the assembly of the microvillar actin bundle by a mechanism that does not prevent monomer association with the preferred end of microfilaments. For instance, villin may stabilize actin filaments by lateral interactions. The functional importance of the carboxy-terminal F-actin binding site in such a mechanism is stressed by the fact that it is required for the formation of F-actin rodlets in cytochalasin D-treated cells. Finally, our data further emphasize the observations that the effects of cytochalasin D in living cells can be modulated by actin-binding proteins. PMID- 8408304 TI - Characterization of calpain II in the retina and photoreceptor outer segments. AB - Calpain II was purified to apparent homogeneity from bovine neural retinas. It was found to be biochemically similar to brain calpain II, purified by the same procedure, with respect to: subunit mobility in SDS-polyacrylamide gel electrophoresis; Ca2+ sensitivity; inhibition by calpeptin and other cysteine protease inhibitors; and optimal pH. Semithin cryosections were immuno-labeled with antibodies specific for the catalytic subunit of calpain II. Calpain II was detected in most layers of the retina, with the most pronounced label present in the plexiform layers (synaptic regions) and the photoreceptor outer segments. In dark-adapted retinas, the label was distributed throughout the outer segments. In light-adapted retinas, outer segment labeling was concentrated in the connecting cilium, and the inner segments were labeled. A partially pure preparation of calpain II from isolated rod outer segments was found to have the same biochemical characteristics as calpain II prepared in the same way from the whole retina. The enzyme was distributed fairly evenly between the cytosolic and cytoskeletal fractions of isolated rod outer segments. Immunoblots of the rod outer segment cytoskeleton were used to determine the susceptibility of known components of the actin-based cytoskeleton to proteolysis by calpain II in vitro. Actin was not proteolyzed at all, alpha-actinin was only slowly degraded, but myosin II heavy chain was rapidly proteolyzed. Actin filaments have been shown previously to be associated with myosin II and alpha-actinin in a small domain within the connecting cilium, where they play an essential role in the morphogenesis of new disk membranes. The localization of calpain II in the connecting cilium after light exposure, combined with the in vitro proteolysis of myosin II, suggests that calpain II could be involved in light-dependent regulation of disk membrane morphogenesis by proteolysis of myosin II. PMID- 8408305 TI - Protein localization to the nucleolus: a search for targeting domains in nucleolin. AB - Nucleolin, a major nucleolar phosphoprotein, is presumed to function in rDNA transcription, rRNA packaging and ribosome assembly. Its primary sequence was highly conserved during evolution and suggests a multi-domain structure. To identify structural elements required for nuclear uptake and nucleolar accumulation of nucleolin, we used site-directed mutagenesis to introduce point- and deletion-mutations into a chicken nucleolin cDNA. Following transient expression in mammalian cells, the intracellular distribution of the corresponding wild-type and mutant proteins was determined by indirect immunofluorescence microscopy. We found that nucleolin contains a functional nuclear localization signal (KRKKEMANKSAPEAKKKK) that conforms exactly to the consensus proposed recently for a bipartite signal (Robbins, J., Dilworth, S.M., Laskey, R.A. and Dingwall, C. (1991) Cell 64, 615-623). Concerning nucleolar localization, we found that the N-terminal 250 amino acids of nucleolin are dispensible, but deletion of either the centrally located RNA-binding motifs (the RNP domain) or the glycine/arginine-rich C terminus (the GR domain) resulted in an exclusively nucleoplasmic distribution. Although both of these latter domains were required for correct subcellular localization of nucleolin, they were not sufficient to target non-nucleolar proteins to the nucleolus. From these results we conclude that nucleolin does not contain a single, linear nucleolar targeting signal. Instead, we propose that the protein uses a bipartite NLS to enter the nucleus and then accumulates within the nucleolus by virtue of binding to other nucleolar components (probably rRNA) via its RNP and GR domains. PMID- 8408306 TI - ors12, a mammalian autonomously replicating DNA sequence, associates with the nuclear matrix in a cell cycle-dependent manner. AB - Origin enriched sequence ors8 and ors12, have been isolated previously by extrusion of nascent CV-1 cell DNA from replication bubbles at the onset of S phase. Both have been shown to direct autonomous DNA replication in vivo and in vitro. Here, we have examined the association of genomic ors8 and ors12 with the nuclear matrix in asynchronous and synchronized CV-1 cells. In asynchronously growing cells, ors8 was found to be randomly distributed, while ors12 was found to be enriched on the nuclear matrix. Using an in vitro binding assay, we determined that ors12 contains two attachment sites, each located in AT-rich domains. Surprisingly, in early and mid-S-phase cells, ors12 homologous sequences were recovered mainly from the DNA loops, while in late-S the majority had shifted to positions on the nuclear matrix. In contrast, the distribution of ors8 over the matrix and loop DNA fractions did not change during the cell cycle. By bromodeoxyuridine substitution of replicating DNA, followed by immunoprecipitation with anti-bromodeoxyuridine antibodies and PCR amplification, we demonstrated that ors12 replicates almost exclusively on the matrix in early and mid-S-phase; replicating ors8 was also found to be enriched on the matrix in early S-phase. Chase experiments showed that the ors12 sequences labelled with bromodeoxyuridine in the first 2 hours of S-phase remain attached to the nuclear matrix, resulting in an accumulation of ors12 on the nuclear matrix at the end of the S period. PMID- 8408307 TI - An immunocytochemical analysis of the vacuolar proton pump in Dictyostelium discoideum. AB - Antisera were generated in rabbits against the vacuolar proton pump (V-H(+) ATPase) purified from Dictyostelium discoideum. The antisera inhibited V-H(+) ATPase but not F1-ATPase activity and immunoprecipitated and immunoblotted only the polypeptide subunits of the V-H(+)-ATPase from cell homogenates. Immunocytochemical analysis of intact cells and subcellular fractions showed that the predominant immunoreactive organelles were clusters of empty, irregular vacuoles of various sizes and shapes, which corresponded to the acidosomes. The cytoplasmic surfaces of lysosomes, phagosomes and the tubular spongiome of the contractile vacuole also bore the pump antigen. The lumina of multivesicular bodies were often stained intensely; the internalized antigen may have been derived from acidosomes by autophagy. Antibodies against V-H(+)-ATPases from plant and animal cells cross-reacted with the proton pumps of Dictyostelium. Antisera directed against the V-H(+)-ATPase of Dictyostelium decorated a profusion of small vacuoles scattered throughout the cytoplasm of hepatocytes, epithelial cells, macrophages and fibroblasts. The pattern paralleled that of the endocytic and acidic spaces; there was no clear indication of discrete acidosomes in these mammalian cells. We conclude that the V-H(+)-ATPase in Dictyostelium is distributed among diverse endomembrane organelles and is immunologically cross reactive with the proton pumps on endocytic vacuoles in mammalian cells. PMID- 8408308 TI - Effects of axotomy on protein synthesis in the rat hypoglossal nucleus: examination of the influence of local recycling of leucine derived from protein degradation into the precursor pool. AB - The quantitative autoradiographic L-[1-14C] leucine method for the determination of regional rates of cerebral protein synthesis (lCPSleu) requires knowledge of the degree of recycling of leucine derived from protein degradation into the precursor pool for protein synthesis, which can be evaluated by measuring lambda i, the steady-state ratio of the leucine-specific activity in the precursor amino acid pool (tRNA-bound leucine) to that of the arterial plasma. To define the changes in lCPSleu during regeneration of the hypoglossal nerve, we examined the effects of axotomy on the value of lambda i. Because the concentration of tRNA bound leucine in the hypoglossal nucleus is too low to measure, we measured the equivalent ratio for the total acid-soluble pool (psi i) and applied the linear relationship between lambda and psi found in the whole brain to calculate a value of lambda i in the ipsilateral and contralateral hypoglossal nuclei of 22 adult female rats 2, 18, 35, and 60 days after unilateral hypoglossal axotomy. Statistically significant but quantitatively inconsequential effects of axotomy on values of psi i and lambda i were found. Therefore, the mean value for lambda i (0.64) of the left and right hypoglossal nuclei in all 22 axotomized rats was used to calculate lCPSleu. In a separate group of 15 unilaterally axotomized rats, lCPSleu was determined by the autoradiographic technique; lCPSleu was increased on the axotomized side by 23% on day 2, 30% on day 18, and 13% on day 35. By postaxotomy day 60, lCPSleu had returned to normal. PMID- 8408309 TI - Fastigial stimulation increases ischemic blood flow and reduces brain damage after focal ischemia. AB - Electrical stimulation of the cerebellar fastigial nucleus (FN) increases CBF and reduces brain damage after focal ischemia. We studied whether FN stimulation "protects" the brain from ischemic damage by increasing blood flow to the ischemic territory. Sprague-Dawley rats were anesthetized (halothane 1-3%) and artificially ventilated through a tracheal cannula inserted transorally. CBF was monitored by a laser-Doppler probe placed over the convexity at a site corresponding to the area spared from infarction by FN stimulation. Arterial pressure (AP), blood gases, and body temperature were controlled, and the electroencephalogram (EEG) was monitored. The stem of the middle cerebral artery (MCA) was occluded. After occlusion, the FN was stimulated for 60 min (100 microA; 50 Hz; 1 s on-1 s off) while AP was maintained at 97 +/- 11 mm Hg (mean +/- SD) by controlled hemorrhage. Rats were then allowed to recover, and infarct volume was determined 24 h later in thionin-stained sections. In unstimulated rats (n = 7), proximal MCA occlusion reduced CBF and the amplitude of the EEG. One day later, these rats had infarcts involving neocortex and striatum. FN stimulation after MCA occlusion (n = 12) enhanced CBF and EEG recovery [61 +/- 34 and 73 +/- 43%, respectively at 60 min; p < 0.05 vs. unstimulated group; analysis of variance (ANOVA)] and reduced the volume of the cortical infarct by 48% (p < 0.05). In contrast, hypercapnia (PCO2 = 64 +/- 4; n = 7) did not affect CBF and EEG recovery or infarct volume (p > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408310 TI - Reductions in focal ischemic infarctions elicited from cerebellar fastigial nucleus do not result from elevations in cerebral blood flow. AB - To determine whether the neuroprotection elicited from electrical stimulation of the cerebellar fastigial nucleus (FN) is attributable to the elevation in regional cerebral blood flow (rCBF), we compared the effects in spontaneously hypertensive rats of stimulation of the rostral ventrolateral medulla (RVL) or FN on (a) a focal ischemic lesion produced by middle cerebral artery (MCA) occlusion, and (b) the changes in rCBF, measured by laser-Doppler flowmetry for 1.5 h, over regions corresponding to the ischemic core (parietal cortex), penumbra (occipital cortex), and nonischemic area (contralateral parietal cortex). Stimulation of FN for 1 h following MCA occlusion reduced infarction 24 h later by 52%. Stimulation of RVL was ineffective. Changes in the lesion were confined to the penumbra. FN and RVL stimulation comparably and significantly increased rCBF up to 185% in unlesioned animals. Following MCA occlusion, stimulation of FN or RVL and hypercarbia failed to elevate rCBF in the ischemic area but did so in the nonischemic area, even though in the same animals only FN stimulation reduced infarction 24 h later. We conclude that (a) the neuroprotection elicited from FN is not the result of an increase in rCBF but results from another mechanism, possibly reduction of metabolism in penumbra, and (b) the pathways mediating central neurogenic vasodilation and neuroprotection are, in part, distinct. PMID- 8408311 TI - Changes in brain capillary diameter during hypocapnia and hypercapnia. AB - Since changes in the surface area of capillaries may be relevant to capillary exchange, the distensibility of brain capillaries was investigated. Brain capillary diameters were measured after perfusion fixation of brain tissue at a constant perfusion pressure during hypo- or hypercapnia. Sections were embedded, stained, and analyzed by light microscopy. The results showed significant differences in mean capillary diameter between the hypocapnic and the hypercapnic group. In the eight brain structures analyzed, capillary diameters were always larger in the hypercapnic group. Mean capillary diameter was 4.93 +/- 0.29 microns in the hypocapnic group and 5.91 +/- 0.10 microns in the hypercapnic group (means +/- SD). We conclude that brain capillaries exhibit a moderate degree of distensibility. Variations in the precapillary pressure of microvessels may therefore influence both capillary flow and capillary surface area. PMID- 8408312 TI - In vitro model of hypoxia: basic fibroblast growth factor can rescue cultured CNS neurons from oxygen-deprived cell death. AB - We established an in vitro hypoxia model and investigated the protective effect of basic fibroblast growth factor (bFGF) against neuronal cell death caused by hypoxia. Hippocampal neurons obtained from rats on embryonic day (E) 17 and 20 and on postnatal day (P) 4 were cultured for 6-24 h in an oxygen-deprived state. This in vitro hypoxia study showed that the cultured neurons were sensitive to the oxygen deprivation. The cultured P4 rat hippocampal neurons seemed to be weaker in the hypoxia condition than those of E17 and E20 rats, suggesting that the cultured postnatal cells might be sensitive to hypoxia. bFGF, but not nerve growth factor, prevented the neuronal cell death caused by hypoxia in a dose dependent manner. PMID- 8408313 TI - DFMO reduces cortical infarct volume after middle cerebral artery occlusion in the rat. AB - Ornithine decarboxylase (ODC), a key enzyme in polyamine biosynthesis, is induced in ischemic tissue and may mediate vasogenic edema and delayed neuronal death. We determined the effects of alpha-difluoromethylornithine (DFMO), a specific inhibitor of ODC, on infarct size and ODC activity in a rat model of transient focal ischemia. DFMO blocked the ischemia-induced increase in ODC and significantly reduced infarct volumes by 57-45%, depending upon the treatment regimen. These studies suggest that polyamine metabolism plays a role in the development of cerebral infarction after focal ischemia and that DFMO may be useful in limiting injury after a stroke. PMID- 8408314 TI - Analysis of covariance in statistical parametric mapping. PMID- 8408315 TI - Instability of localization of cerebral blood flow activation foci with parametric maps. PMID- 8408316 TI - Localized dynamic changes in cortical blood flow with whisker stimulation corresponds to matched vascular and neuronal architecture of rat barrels. AB - The hypothesis that functional groups of neurons in whisker barrels are linked to a modular organization of cortical vessels was tested. Endovascular casts demonstrated cortical capillary networks resembling the whisker barrel pattern that were fed from the middle cerebral artery. In histological sections, dense capillaries apparently were confined to single barrels and were supplied by one or a few penetrating arterioles. The barrel field in cortical layer IV was localized in relation to surface arteriovenous patterns. Living vessels were imaged through a closed cranial window under anesthesia with a fluorescence microscope and SIT or ICCD cameras. After intracarotid injections of fluorescein isothiocyanatedextrans, saline, or 3 microns latex beads, changes in arteriolar diameter, arteriovenous transit times (AVTTs), and bead velocities were measured. When row C whiskers were stroked at 4-5 Hz for 1 min, blood flow increased in arterioles that supplied contralateral row C barrels as demonstrated by postmortem histology. AVTTs slowed significantly in vessels supplying adjacent cortex. We hypothesize that cerebral vascular units supply individual whisker barrels and are functionally linked to them for precise focal regulation of cerebral blood flow. PMID- 8408317 TI - Differential transcription and translation of immediate early genes in the gerbil hippocampus after transient global ischemia. AB - Excitotoxic activation of glutamate receptors is thought to be a key event for the molecular pathogenesis of postischemic delayed neuronal death of CA-1 neurons in the gerbil hippocampus. Glutamate receptor stimulation also causes induction of transcription factors that belong to the class of immediate early genes. We examined the expression of six different immediate early genes in the gerbil hippocampus after transient global ischemia. Comparative analysis of c-fos and Krox-24 expression was carried out in the same animals at the transcriptional and translational level by in situ hybridization and immunocytochemistry. Postischemic synthesis of four additional immediate early gene (IEG)-encoded proteins (FOS-B, c-JUN, JUN-B, and JUN-D) was investigated by immunocytochemistry at recirculation intervals between 1 and 48 h. After 5 min of ischemia, transcription of c-fos and Krox-24 mRNA was induced in all hippocampal subpopulations with peak expression at 1 h after recirculation. In vulnerable CA 1 neurons, increased transcription of c-fos and Krox-24 was not followed by translation into protein. Induction of immediate early gene-encoded proteins was restricted to neuronal populations less vulnerable to brief ischemia and identified neurons that are targets of glutamate receptor-mediated neurotoxicity but that are destined to survive. Our data indicate an asynchronous synthesis and persistence of individual IEG-encoded proteins in these neurons. The staggered induction implies that combinatorial changes of transcription factors allow a differential postischemic regulation of target gene expression both spatially and over time. PMID- 8408318 TI - MK-801 (dizocilpine) protects the brain from repeated normothermic global ischemic insults in the rat. AB - We investigated the neuroprotective potential of MK-801 (dizocilpine), a noncompetitive N-methyl-D-aspartate (NMDA) antagonist, in the setting of three 5 min periods of global cerebral ischemia separated by 1-h intervals in halothane anesthetized rats. Each ischemic insult was produced by bilateral carotid artery occlusions plus hypotension (50 mm Hg). Brain temperature was maintained at normothermic levels (36.5-37.0 degrees C) throughout the experiment. MK-801 (3 mg/kg) (n = 6) or saline (n = 6) was injected intraperitoneally 45 min following the end of the first ischemic insult. Following 7-day survival, quantitative neuronal counts of perfusion-fixed brains revealed severe ischemic damage in hippocampal CA1 area, neocortex, ventrolateral thalamus, and striatum of untreated rats. By contrast, significant protection was observed in MK-801 treated rats. In area CA1 of the hippocampus, numbers of normal neurons were increased 11- to 14-fold by MK-801 treatment (p < 0.01). The ventrolateral thalamus of MK-801-treated rats showed almost complete histologic protection, and neocortical damage was reduced by 71% (p < 0.01). The degree of MK-801 protection of striatal neurons was less complete than that seen in other vulnerable structures, amounting to 63% for central striatum (p = 0.02, Mann-Whitney U test) and 48% in the dorsolateral striatum (NS). A repeated-measures analysis of variance demonstrated a highly significant overall protective effect of MK-801 treatment (F1,10 = 37.2, p = 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408319 TI - Flunarizine blocks elevation of free cytosolic calcium in synaptosomes following sustained depolarization. AB - Gerbil cerebral cortical synaptosomes loaded with the fluorescent calcium probe FURA-2 were used to study depolarization-induced presynaptic cytosolic free calcium concentration, as an in vitro model of cerebral ischemia. The depolarization-induced increase in intrasynaptosomal cytosolic free calcium concentration is not sodium-dependent or sodium channel-dependent and may be due to an influx of extrasynaptosomal calcium resulting from a cadmium- and omega conotoxin-sensitive, nickel-, nifedipine-, and nimodipine-insensitive voltage regulated channel. The depolarization-induced increase in intrasynaptosomal free cytosolic calcium concentration is also inhibited by flunarizine, a calcium antagonist that has protective effects in animal models of cerebral anoxia and ischemia. Our results suggest that presynaptic calcium uptake following depolarization may be mediated in part by an N-type channel. Flunarizine may block presynaptic calcium accumulation, in part, by blocking this N-type channel; this blockade may be just one of several mechanisms by which flunarizine exerts protective effects following cerebral ischemia. PMID- 8408320 TI - Brain calcium metabolism in hypoglycemic coma. AB - The present experiments were designed to provide information on brain calcium metabolism during hypoglycemic coma. We specifically wished to evaluate changes in extracellular calcium concentration (Ca2+e) during prolonged hypoglycemic coma and recovery and to assess whether Ca2+e falls to similar values during hypoglycemia and ischemia. To that end, Ca2+e and K+e in neocortical tissue were recorded by ion-sensitive microelectrodes during hypoglycemic coma of 30 min duration and during 15 min of recovery. Cardiac arrest ischemia was induced either at the end of the period of hypoglycemia or after 15 min of recovery. Hypoglycemic coma, as reflected by a DC potential shift and by cellular release of K+, was accompanied by a sustained decrease in Ca2+e from approximately 1.2 to approximately 0.02 mM, i.e., to approximately 1% of control. Infusion of glucose was followed by a biphasic recovery of Ca2+e, starting within 2 min of infusion. During the first phase, completed within the initial 3-4 min, Ca2+e rose to about 25% of control. During the second phase, Ca2+e slowly increased toward normal within 25-30 min. Ischemia, when induced at the end of the period of hypoglycemia, was accompanied by a rise in Ca2+e to about 0.1 mM, i.e., about 10% of control. A similar value was recorded when ischemia was induced after 15 min of recovery following a 30-min hypoglycemic coma. Although the present results do not give information on Ca2+i during hypoglycemic coma, it is tempting to conclude that partial preservation of the nucleoside triphosphate stores, and absence of acidosis, allow some binding and sequestration of the calcium entering the cell.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408321 TI - Ketamine alters calcium and magnesium in brain tissue following experimental head trauma in rats. AB - We previously reported that the N-methyl-D-aspartate receptor antagonists dizocilpine maleate and ketamine improved the neurological severity score (NSS) after head trauma in rats. Other investigators have reported increased calcium and decreased magnesium following head trauma in untreated rats. The present study was designed to determine whether ketamine influences the concentrations of calcium and magnesium in brain tissue following head trauma. Eighty-six male Sprague-Dawley rats (180 +/- 15 g) were divided into eight groups. Groups A (no head injury) and C (head injury) received no treatment. Groups B (no head injury) and D-H (head injury) received ketamine. In groups D, E, and F, ketamine, 180 mg/kg i.p., was given 1, 2, and 4 h after head trauma, respectively. In groups G and H, ketamine, 120 and 60 mg/kg, respectively, was given 1 h after head trauma. After we killed the rats at 48 h, cortical slices were taken to measure tissue calcium and magnesium content by the inductively coupled plasma atomic emission spectroscopy method. In the contused hemispheres, calcium increased and magnesium decreased (p < 0.0001). Among the head-injured groups, the increase in brain tissue calcium was smaller in groups receiving 60 mg/kg of ketamine at 1 h or 180 mg/kg of ketamine at 1, 2, or 4 h than in the group not receiving ketamine. The decrease in brain tissue magnesium was smaller in the groups receiving 180 mg/kg of ketamine at 1 and 2 h than in the group not receiving ketamine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408322 TI - Local NADPH-diaphorase neurons innervate pial arteries and lie close or project to intracerebral blood vessels: a possible role for nitric oxide in the regulation of cerebral blood flow. AB - Electrical stimulation of perivascular nerves induced a relaxation of endothelium denuded cat pial arteries that was significantly reduced by nitric oxide (NO) synthase inhibition, indicating that NO was involved in the neurogenic relaxation of these vessels. Histochemical staining of the pial arteries for NADPH diaphorase (NADPH-d), used as a marker for NO synthase, showed positive nerve fibers in the adventitial layer. Interestingly, in some restricted areas stained neuronal cell bodies were also observed. These neurons were scattered or distributed in small groups in a ganglion-like manner, and they sent fibers to the vessel wall. No NADPH-d-positive nerve fibers or cell bodies were detected in forelimb, pulmonary, or coronary arteries. Within the brain parenchyma, blood vessels also showed positive fibers around their walls. These fibers were organized in a branching pattern and presented varicosities. NADPH-d-positive neurons were found in the proximity of the intracerebral vascular profiles, sending processes to the vessels and/or being directly apposed to their wall. The neurovascular contacts were preferentially located close to the interface between the cerebral cortex and white matter. The anatomical relationship between NADPH-d positive neurons and fibers and the cerebral blood vessels, together with the participation of NO in the neurogenic relaxation of pial arteries, suggests that NO is involved in the regulation of cerebral blood flow. PMID- 8408323 TI - Time course of hippocampal glucose utilization and persistence of parvalbumin immunoreactive neurons after ibotenic acid-induced lesions of the rat dentate area. AB - The effects of ibotenic acid induced lesions of the dentate gyrus on hippocampal glucose utilization and parvalbumin-positive neurons were evaluated in male Wistar rats. Ibotenic acid was injected in the right dorsal dentate gyrus. Quantification of glucose utilization was performed 3 days, 3 weeks, or 3 months after the lesion using the 14C-2-deoxyglucose method. Nissl-stained sections and sections stained for acetylcholinesterase were used as references for anatomical delineation of the hippocampal cytoarchitecture. Additional sections were stained for parvalbumin. The results revealed widespread reductions of glucose utilization in all layers and sectors of the hippocampus in the ipsilateral lesioned hemisphere and also in the nonlesioned contralateral hemisphere. The reductions occurred as early as 3 days after the lesion and persisted up to 3 months. In neither hippocampal structure did glucose utilization return to control levels. Immunohistochemical visualization of parvalbumin-containing neurons revealed that these putatively inhibitory neurons persisted in the otherwise granule-cell-depleted area. The data show that interruption of the excitatory trisynaptic pathway from the entorhinal cortex to the CA1 at the level of the dentate gyrus affects hippocampal glucose utilization irreversibly and uniformly. Since some inhibitory neurons seem to survive the ibotenic acid lesion, we suggest that the reductions of hippocampal glucose utilization reflect an imbalance in favor of inhibitory neurons in the ipsilateral hippocampus after the lesion, which manifests also in the contralateral hemisphere via the commissural pathways. PMID- 8408324 TI - [Celioscopic treatment of ectopic pregnancy. Results apropos of a series of 109 cases]. AB - We report our experience concerning the laparoscopic treatment of ectopic pregnancy (EP). One-hundred and nine women with EP were treated in our department over a 4 year period, between February 1988 (date of our first laparoscopic surgery for EP) and December 1991. Twenty-two these women underwent laparotomy and the remaining 87 laparoscopic surgery alone. Four therapeutic failures were noted in women treated by laparoscopy. Our results are compared with those of other series and the indications, as well as the modalities of laparoscopic treatment are detailed. It is concluded that laparoscopic surgery of EP is a reliable method which must always be considered, except for a few rare indications. PMID- 8408325 TI - [Shouldice's operation: can results in a general surgical unit be the same as those in a highly specialized surgical unit?]. AB - In the course of 5 years 1803 groin hernia operations have been carried out, 1640 of them Shouldice operations, in a General Surgery Service. Post operation follow up of patients operated with Shouldice technique has shown 0.72% recurrence in patients checked for over a year, and 2.13% complications. Correct applications of this technique gives the same results as those in specialized centres. PMID- 8408326 TI - [Cervical cystic lymphangiomas in adults]. AB - Cystic hygromas, infrequent in children, are even rarer in adults where they are generally unexpected operative findings. Now, with the progress in modern medical imaging, the diagnosis can be suggested before operation. Analysis of the embryogenesis helps to explain the different anatomical and clinical forms and the relations of the cystic hygroma with neighboring tissues, particularly of venous origin. 9 cases of cystic hygroma of the neck in adults are reported and discussed with 26 other well documented cases from the literature. The diagnosis can be confirmed only after operation and histological examination. Surgical removal is technically less difficult in adults than in children and recurrences are rare. PMID- 8408327 TI - [Gritti's amputation in vascular surgery]. AB - Gritti's amputation was performed in 35 patients, mean age 71 years, with vascular disease of leg. There was no immediate postoperative mortality. Of twenty patients fitted with an appliance using a specific technique, only one had a poor functional result. The results obtained in this preliminary series were compared with those after amputations and knee disarticulations. They showed that Gritti's amputation should be considered for patients with arteritis when a functional knee is impossible to conserve. PMID- 8408328 TI - [Preservation of a foot by shortening tibial osteotomy after a circumferential electric burn to the leg]. AB - A patient presented with circumferential loss of skin substance of lower quarter of leg. The patient's advanced age and the poor local conditions could have produced an aleatory result for a flap graft. It was therefore decided to perform a shortening tibial osteotomy as a conservative surgical procedure. PMID- 8408329 TI - [Non-traumatic perforation of the small intestine]. AB - An overview of a series of patients treated for peritonitis over the last 5 years showed that in 64 cases (6.8%) the infection was due to a non traumatic perforation of small intestine. The predominant aetiology was typhoid fever (39 cases), other causes for the peritonitis were perforation of an abdominal diastasis (10 cases) or a Meckel's diverticulum (8 cases), and perforation due to an acute ileitis (2 cases), a non Hodgkins malignant lymphoma (2 cases), a necrotizing enteritis (2 cases) and a jejunal tuberculoma (1 case). The surgical attitude to be adopted for repair of the perforated loop is dependent on the aetiology and the degree of peritoneal sepsis. Enterostomy should be performed as a safety measure in patients with perforation due to typhoid fever. PMID- 8408330 TI - [Inflammatory syndrome, anicteric cholestasis and hepatic adenoma]. AB - An unusual form of adenoma of left lobe of liver was detected fortuitously in a young woman with an inflammatory syndrome and an anicteric cholestasis. In this case, a positive correlation existed between the adenoma and the treatment with estrogens, the disappearance of the biologic anomalies after exeresis of the adenoma being suggestive of an increased production of fibrinogen by hepatocytes. The cholestasis was probably the result of pressure exerted by the adenoma. Excision of the adenoma was justified by the risk of hemorrhagic complications and the possibility of malignant changes in the tumor. PMID- 8408331 TI - [Cancer of the breast in males]. AB - Between 1982 and 1990, 1027 women and 13 men were operated upon for cancer of the breast. Analysis failed to show any difference with sex for age, localization or symptomatology. Distribution by stage, as defined by the U.I.C.C., showed a higher incidence of stage III in the men and of stage II in the women. Treatment consisted of a modified Patey type radical mastectomy in the men, a more limited intervention being applied in 20% of the women. After a mean follow up of 62 months, none of the men had died of their breast cancer. The recurrence rate was 16%. Results in this series and in those reported in the literature emphasize the decisive prognostic value of primary surgical treatment. Complementary hormone therapy with tamoxifen is particularly indicated, since 90% of the patients had positive estrogen hormone receptors. PMID- 8408332 TI - [Treatment of sub-levator rectocele by transrectal approach. Value of the automatic stapler with linear clamping]. AB - Surgical treatment of rectocele causing outlet blockage is still a subject of controversy. A retrospective study of eight rectocele repairs endorectally using a surgical stapling device done over a two years period was performed. The most common indication was constipation. All patients underwent in the preoperative period complete physiologic examination including defecography, anorectal manometry, electromyography and colonic transit studies. All patients underwent colonoscopy to exclude an obvious physical disorder. There was no postoperative morbidity and the mean hospital stay was four days. Good functional results were obtained in seven patients with a one year follow-up. Endorectal resection using a stapling device is both a simple and effective technique to treat rectocele associated with difficulty in evacuation. PMID- 8408333 TI - [Treatment of complex loss of substance in acute leg injuries]. AB - Solving the problems inherent in the treatment of Gustillo type III open fractures of leg requires the intervention of multiple surgical disciplines: orthopedic, plastic and vascular. Consensus exists with respect to many therapeutic principles, but several points remain the subject of debate. Based on an extensive literature review, the indications, results and prognosis for both skin covery and bone reconstruction are defined as precisely as possible. Early treatment is essential, the loss of soft tissue substance being repaired within the first two weeks, particularly if the use of a free flap is envisaged. Two stage treatment, cutaneous and bony, offers the advantage of safety, with additional benefits if conventional grafts are applied within 1 to 2 months. Overall therapeutic results are dependent on good interdisciplinary team work and, to an increasing extent, on the urgent participation of plastic surgeons experienced in microsurgical techniques. PMID- 8408334 TI - [Cystic lymphangioma of the small omentum]. PMID- 8408336 TI - Outcomes analysis and the practice of medicine. PMID- 8408335 TI - [Rupture of a woven dacron prosthesis used for treatment of a large hernia]. PMID- 8408337 TI - Paper-bag rebreathing. PMID- 8408338 TI - Role models. PMID- 8408339 TI - Proctological truth. PMID- 8408340 TI - A complication during bronchoscopy. PMID- 8408341 TI - New devices for coronary revascularization. AB - A variety of instruments, including atherectomy catheters, intra-arterial stents, and lasers, have been developed in order to outdo balloon angioplasty with regard to procedural success, complications, and restenosis. Controlled trials are needed to assess the newcomers, but it is likely that each type of device will find its own clinical niche rather than supersede the others. PMID- 8408342 TI - Severe hypertension in a young patient. PMID- 8408343 TI - Books doctors read. PMID- 8408344 TI - Acute illness and subcutaneous nodules. PMID- 8408345 TI - Revitalizing primary care: a 10-point proposal. AB - Even if all Americans had medical insurance, many of them still would not have access to adequate medical care, largely because of the lack of primary care physicians. Increasing their numbers is one aspect of health care reform in which the medical community can have a resounding impact. Specific recommendations for shaping the next generation of physicians are offered. PMID- 8408346 TI - Diagnostic and therapeutic options in endometriosis. PMID- 8408347 TI - Can control of infectious diseases be sustained? PMID- 8408348 TI - Legionnaires' disease: new understanding of community-acquired pneumonia. PMID- 8408349 TI - Aortic blebs: an indication for urgent surgery. AB - A blister on an aortic aneurysm represents the final stage before rupture, regardless of patient symptoms or aneurysm size. The mechanisms of rupture are unclear but probably involve defects in the elastin and collagen matrices. PMID- 8408350 TI - Headaches in a pregnant woman with a history of preeclampsia. PMID- 8408351 TI - The challenge of breaking bad news. PMID- 8408352 TI - Facing the human face of disease. PMID- 8408353 TI - A woman with shock and normal cardiac output after acute MI. PMID- 8408354 TI - One procedure or two? PMID- 8408355 TI - Night sweats. PMID- 8408356 TI - Ticlopidine and stroke prevention. PMID- 8408357 TI - The 'sticky' problem of Staphylococcus epidermidis sepsis. PMID- 8408358 TI - Bronchial asthma: diagnostic and treatment issues. AB - The increasing morbidity and mortality associated with asthma may, in part, be due to inadequate treatment. Step-care management aims to improve control by addressing both the bronchospastic and inflammatory components of the disease. beta-Adrenergic agonists are central to alleviating symptoms, whereas corticosteroids may reduce inflammation and associated airway hyperresponsiveness. PMID- 8408359 TI - Identifying depression: the significance of anhedonia. PMID- 8408360 TI - Ascites and malnutrition in a chronic dialysis patient. PMID- 8408361 TI - The hypercoagulability of polycythemia vera. AB - Thrombosis is the most common cause of death in patients with this myeloproliferative syndrome. Phlebotomy, hydroxyurea, and antiplatelet agents are the mainstays of therapy but may not stave off a lethal embolism. PMID- 8408362 TI - Pigmented lesions in the elderly. PMID- 8408363 TI - Toward rational treatment of scleroderma. PMID- 8408364 TI - Methionine metabolism: a window on carcinogenesis? AB - Recent experimental evidence links changes in methionine metabolism to the onset and progression of cancer. Aberrant methylation reactions and polyamine synthesis may alter genome stability, gene expression, and cell proliferation. PMID- 8408365 TI - Prevalence of childhood depression: results of the first study in Spain. AB - Prevalence of depressive disorders was studied in 6432 children aged 8-11 years, selected by exhaustive random sampling in four Spanish cities and two rural areas. The number of non-participants was 38 (0.6%). Selection was by a two-stage procedure. At stage I, the Children's Depression Inventory was used to identify a sample for more intensive interviewing. At stage II, children were individually evaluated using a semi-structured interview, the Children's Depression Rating Scale-Revised (CDRS-R). CDRS-R and DSM-III diagnostic criteria were used to define caseness. 1.8% of children met the criteria for major depression, with a further 6.4% being diagnosed as having dysthymic disorder. PMID- 8408366 TI - Developmental normative data on the test of variables of attention (T.O.V.A.). AB - Developmental normative data for 775 children aged 6-16 are presented for the Test of Variables of Attention (T.O.V.A.), a 23-minute fixed-interval visual Continuous Performance Test with minimal language demands and no left-right discrimination. The target is presented on 22.5% and 77.5% of the trials during the first and second halves, respectively, T.O.V.A. indices include omission and commission errors, response time means and standard deviations, and anticipatory responses. Attention and impulse control developed in a non-linear manner, changing rapidly in early childhood and leveling off during later childhood and adolescence. PMID- 8408367 TI - A longitudinal study of neural maturation and early mother-infant interaction: a research note. AB - The development of postural control of the head and of looking towards and looking away from the mother is described; the age relationship between attainment of postural head control and looking towards the mother is investigated and the relation between smiling and "pleasure" vocalizations to looking towards and away is examined. Twelve normal, full-term mother-infant pairs participated in this home-based study. En face interaction was video-taped for maximally 15 min at 3, 6, 9, 12, 15 and 18 (or 21) weeks. We conclude that postural head control constitutes an important prerequisite for the emergence of socially directed behaviours during en face interaction. PMID- 8408368 TI - The effectiveness of cognitive self-management as an adjunct to a behavioural intervention for childhood obesity: a research note. AB - This study investigated the effectiveness of cognitive self-management training as an adjunct to the behavioural management of childhood obesity. Twenty-seven overweight children aged 7-13 years were randomly assigned to either behavioural management plus relaxation placebo or a combined behavioural-cognitive self management approach. Evaluations following the eight treatment sessions revealed a significant reduction in percentage overweight for children in both experimental groups and improvements were maintained at 3- and 6-month follow ups. Both conditions were also effective in reducing the number of high-risk foods consumed. No difference in outcome was found between treatments at the post treatment assessment or 3- and 6-month follow-ups. Although a reduction in percentage overweight of around 9% was found for both procedures, subjects in general remained considerably overweight. PMID- 8408369 TI - The Vineland Adaptive Behavior Scale in a sample of normal French Children: a research note. AB - The Vineland Adaptive Behavior scale (survey form) was used in a sample of 151 normal children under age 18. Standardized mean scores of French children were comparable to those of the American normative sample. From the age of 6 onwards, French children scored consistently lower in the Daily Living Skills domain though the magnitude of this difference remained moderate. While the overall findings support the cross-cultural stability of the psychometric properties of this instrument, attention is drawn to potential problems in the use of the Vineland scales, with special reference to autistic samples. PMID- 8408370 TI - Annotation: the preterm infant: psychological issues in childhood. PMID- 8408371 TI - An epidemiological study of disorders in late childhood and adolescence--I. Age- and gender-specific prevalence. AB - Developmental aspects of psychiatric disorders may be inferred from patterns of age differences in prevalence. Age-specific prevalences are provided for nine disorders in a general population sample of ages 10-20. Age and gender patterns for several disorders suggest developmental stage-associated risks. These include oppositional disorder in both genders and conduct disorder and major depression in girls. Major depression shows a pattern suggestive of a role for the onset of puberty. The prevalence of one or more disorders did not differ by age or gender. However, the pattern of specific diagnoses varied greatly by both age and gender. PMID- 8408372 TI - An epidemiological study of disorders in late childhood and adolescence--II. Persistence of disorders. AB - The persistence and new onset of psychiatric disorder were studied in a sample of 734 children from the general population. Diagnoses for six of the more prevalent disorders were generated from maternal and child interviews when the children were ages 9-18 and again 2 1/2 years later. Analyses demonstrated substantial levels of diagnostic persistence over the 2 1/2 year period for all diagnoses except major depression. With few exceptions persistence was roughly equivalent for age and gender subgroups. It is concluded that disorders assessed by structured interview of non-clinical samples of children cannot be dismissed as transitory. PMID- 8408373 TI - Assessment of psychosocial experiences in childhood: methodological issues and some illustrative findings. AB - The development of a new standardised investigator-based interview, PACE (Psychosocial Assessment of Childhood Experiences), for the assessment of acute life events and long-term psychosocial experiences is described. An application of PACE to a sample of 84 children referred to psychiatric clinics and 22 general population controls, is presented. Reliability was assessed using a separate clinic sample of 15 child-parent pairs. The findings showed that PACE has satisfactory reliability and discriminant validity. PMID- 8408374 TI - The effects of conduct disorder and attention deficit in middle childhood on offending and scholastic ability at age 13. AB - The relationship between conduct disorder and attention deficit behaviours in middle childhood (6, 8 and 10 years) and juvenile offending and academic achievement were examined in a birth cohort of New Zealand children. The application of structural equation modelling methods suggested that early behavioural tendencies are related to later developmental outcomes by two highly correlated but distinct developmental progressions. In the first such progression, early conduct disorder behaviours acted as a precursor of future offending patterns but these behaviours were unrelated to later school performance when the correlation between conduct disorder and attention deficit was taken into account. In the second developmental progression early attentional/cognitive behaviours were related to future school performance but were unrelated to the development of antisocial behaviours when the correlations between conduct disorder and cognitive/attentional variables were taken into account. The implications of these findings for validating the distinction between conduct disorder and attention deficit behaviours is discussed and the problems of analysing and explaining the high comorbidity between conduct disorder and attention deficit behaviours are considered. PMID- 8408375 TI - Early hospitalization and disturbances of infant behavior and the mother-infant relationship. AB - The impact of brief early hospitalization was investigated in a controlled natural experiment of 14 children with cleft lip and palate (CLP) who were hospitalized for palatal repair at 9.5 months of age, and another 13 CLP children who were hospitalized at 12.5 months of age; 14 children without clefts who were not hospitalized served as controls. After hospitalization children were more likely to avoid their mothers both following a brief stressful separation at home and in the Strange Situation. It was suggested that CLP children may have learned to cope with stress by shifting the attention away from the mother. PMID- 8408376 TI - Who participates in child sexual abuse research? AB - This paper stems from a prospective case control study of the effects of child sexual abuse and explores issues of sampling bias resulting from non participation in research. Despite similarities in family demographics and in the level of abuse experienced, differences between the study participants and non participants emerged. The more dysfunctional families were less likely to participate, the research was not a passive experience for the families and finally, non-participants were offered less therapy following clinical assessment. The implications of these differences for the main prospective study will be explored. PMID- 8408377 TI - Creating satisfaction: a psychological perspective on stress and coping in families of handicapped children. AB - This paper briefly reviews the literature on stress and coping in families of developmentally handicapped children and proposes an alternative way for conceptualizing some of the psychological processes involved. The approach specifies how threats to certain human needs lead to predictable patterns of appraisal and coping. The usefulness of this alternative is demonstrated for families of autistic children and the theoretical and clinical implications are discussed. PMID- 8408378 TI - Latent inhibition deficits in hyperactive children. AB - Latent inhibition (LI), the retardation of associative learning as a function of prior stimulus exposure, is believed to reflect normal attentional processes. Previous research has demonstrated an attenuated LI effect in acute schizophrenics as well as normal subjects who rate high on schizotypal scales. The present study examined LI in two age groups (66.1 and 77.4 months) of hyperactive and normal children. The younger normal children exhibited LI while the older ones did not. Neither of the hyperactive groups displayed LI. The loss of LI in the hyperactive subjects was explained by proposing that these children are deficient in processing non-salient, irrelevant stimuli. PMID- 8408379 TI - Seclusion on an adolescent unit. AB - We studied the seclusion records of an adolescent unit for a six-year period. Problems in the analysis of typical seclusion data are examined and statistical methods that overcame them are explained. Only a few relationships were found between the seclusion regime and available measures of patient and ward characteristics, though there was great variability in the frequency with which individuals were secluded and the duration of various episodes of seclusion. The average duration of seclusion (31 minutes) was much less than had been reported by other investigators. Furthermore, the durations became shorter over the period of study. The implications of these findings for further research and practice are discussed. PMID- 8408380 TI - Emotional/behavioral problems in clinic and nonclinic children: correspondence among child, parent and teacher reports. AB - This study examined the correspondence among child, parent and teacher reports, in rating emotional and behavioral problems of 98 community nonpatients and 64 clinically referred children (ages 6-13). The mean ratings differed significantly for the three sources and for the two samples, and there were a few significant interactions. Correlations of the scores between sources, especially parents and teachers, were in the moderate range and were not significantly different from one another. Informant correspondence was higher for externalizing than internalizing problems in both patient and nonpatient samples, and was higher for nonpatients than patients. In general, informant agreement was higher for cases characterized by lower family stress and higher child acceptance, among other clinical variables. PMID- 8408381 TI - Quantification of mammalian lignans in biological fluids using gas chromatography with ion mobility detection. AB - A method is presented to quantify selected mammalian lignans in human physiological fluids by gas chromatography (GC) coupled with ion mobility spectrometry (IMS). The use of IMS following GC permitted the selective and sensitive measurement of 2,3-bis(3-hydroxybenzyl)butane-1,4-diol (i.e., enterodiol) and trans-2,3-bis(3-hydroxybenzyl)-gamma-butyrolactone (i.e., enterolactone) concentrations in urine and plasma following dietary supplementation with whole wheat/flaxseed bread high in mammalian lignan precursors. Following six weeks of flaxseed feeding, urinary and plasma levels of enterodiol and enterolactone were elevated, exceeding the amounts found at baseline by a factor of 3-5. The approach to mammalian lignan methodology presented herein provides novel analytical phytochemical procedures for assessing the impact of lignan consumption in human health and disease. PMID- 8408382 TI - Isotope dilution ammonia chemical ionization mass fragmentographic analysis of urinary 3-O-methylated catecholamine metabolites. Rapid sample clean-up by derivatization and extraction of lyophilized samples. AB - We developed a method for simultaneous quantification of the urinary 3-O methylated catecholamine metabolites 3-methoxytyramine, normetanephrine and metanephrine by stable isotope-dilution ammonia chemical ionization mass fragmentography. Prepurification of lyophilized samples was done by simultaneous deconjugation and pentafluoropropionylation, followed by extraction and rederivatization. Compared with our previously described method, based on acid hydrolysis, alkaline extraction, derivatization and electron-impact mass fragmentography, the present method was found to be less laborious, more sensitive and presumably more accurate. New urinary excretion values were established for apparently healthy adults. The present prepurification method may prove applicable for profiling of a variety of naturally occurring mono-, di- and polyamines in biological samples. PMID- 8408383 TI - Gas chromatographic determination of primary and secondary low-molecular-mass aliphatic amines in urine using derivatization with isobutyl chloroformate. AB - A simple routine method for the gas chromatographic determination of methylamine, dimethylamine, ethylamine and methylethylamine in urine is presented. The method is based on a two-phase derivatization procedure with isobutyl chloroformate as reagent. The reaction is quantitative in 10 min. We found no artifact formation of either choline or trimethylamine (dietary amine compounds) or of dimethylethylamine or triethylamine (catalyst amines in the industrial setting). The chromatographic behaviour of the amine carbamates was excellent. The recoveries of methylamine, dimethylamine, ethylamine and methylethylamine in spiked urine samples were 82, 89, 100 and 96%, respectively, and the precision (the relative standard deviation) was 3.6, 1.8, 3.3 and 2.0%, respectively. The method was linear for the studied amine carbamates up to 250 mg/l. The endogenous amine concentrations in urine samples from ten normal subjects were: methylamine, 0.9 mg/l (mean; range 0.3-1.5); dimethylamine, 14.7 mg/l (mean; range 4.6-27.6); ethylamine, 0.8 mg/l (mean; range 0.2-2.3); methylethylamine, less than 0.02 mg/l. PMID- 8408384 TI - Determination of taurine metabolism by measurement of 15N-enriched taurine in cat urine by gas chromatography-mass spectrometry. AB - To understand the biological function of taurine, a study of taurine kinetics in the cat was undertaken. This paper describes a method developed for the accurate determination of 15N-taurine enrichment in cat urine by gas chromatography-mass spectrometry. 15N-Taurine was given to six animals as an oral bolus dose of 20 mg/kg body weight, and the urine was pooled on a daily basis. The hydrolysed or non-hydrolysed urine samples (for total and free taurine, respectively) were directly derivatized without further purification. The N-pentafluorobenzoyl di-n butyl amide derivative obtained was analysed, and the fragment [M-(di-n-butyl amide)]+, carrier of the labelled nitrogen atom, was selectively recorded at m/z 302 (14N-taurine) and m/z 303 (15N-taurine). Calibration curves prepared in hydrolysed and non-hydrolysed urine samples spiked with 15N-taurine gave similar slopes to the calibration curve prepared in water. The average coefficient of variation observed for the mole percent excess in the non-hydrolysed samples was 1.22% (n = 92) and for the hydrolysed urine 1.00% (n = 98). There was no significant difference between free and total taurine enrichment. The half-life of taurine in cat body was found to be 29.3 +/- 2.9 h and 35.0 +/- 1.4 h for free and total taurine, respectively (non-significant). The taurine body pool, calculated by extrapolation of the curve to zero time, had a value of 137 +/- 22 ng/kg and 157 +/- 11 mg/kg for free and total taurine, respectively. PMID- 8408385 TI - Determination of triacylglycerol and cholesterol ester hydroperoxides in human plasma by high-performance liquid chromatography with fluorometric postcolumn detection. AB - Cholesterol ester (ChE) and triacylglycerol (TG) hydroperoxides in human plasma were determined by high-performance liquid chromatography with postcolumn detection with diphenyl-1-pyrenylphosphine. Human plasma was extracted once with n-hexane. 2,6-Di-tert.-butyl-4-methylphenol and N-stearylcinnamide were added to human plasma before extraction as an antioxidation agent and an internal standard, respectively. The detection limits of both ChE and TG hydroperoxides were 1 pmol. The sample size was minimized to 250 microliters for each run. The recoveries of ChE and TG hydroperoxides from fresh plasma were ca. 90 and 80%, respectively. The relative standard deviations (n = 8) of their values in frozen human plasma were 5.4% (ChE hydroperoxides, 298 nM) and 5.7% (TG hydroperoxides, 267 nM). No TG hydroperoxides and 24.5 +/- 9.6 nM (n = 15) ChE hydroperoxides were detected in fresh human plasma. The relative standard deviation (n = 8) of ChE hydroperoxides values in fresh plasma was 5.8% (27.1 nM). PMID- 8408386 TI - Simultaneous determination of cholesterol and cholestanol in human serum by high performance liquid chromatography using 3-(5,6-methylenedioxy-2 phthalimidyl)benzoyl azide as precolumn fluorescent labelling reagent. AB - A fluorescent labelling reagent, 3-(5,6-methylenedioxy-2-phthalimidyl) benzoyl azide, designed for the determination of alcohols by precolumn high-performance liquid chromatography, has been applied to the simultaneous determination of cholesterol and cholestanol in human serum. The reagent reacts with cholesterol and cholestanol at 140 degrees C for 10 min to produce the fluorescent derivatives, which can be separated on a reversed-phase column with acetonitrile ethanol-water (60:35:7.5, v/v) as eluent. The detection limits for cholesterol and cholestanol were 45 and 50 fmol per injection (20 microliters), respectively. The values of cholesterol and cholestanol in normal human sera were 135-212 mg/dl and 137-928 micrograms/dl, respectively. PMID- 8408387 TI - Assay of enzymic O-methylation of catechol oestrogens by high-performance liquid chromatography with coulometric detection. AB - A simple and sensitive method for the determination of guaiaicol oestrogens enzymatically formed from 2- or 4-hydroxyoestradiol, by means of high-performance liquid chromatography with coulometric detection, has been developed. Catechol and guaiacol oestrogens were efficiently separated on a reversed-phase column, using 0.5% ammonium phosphate buffer (pH 3.0)-acetonitrile (59:41, v/v) as the mobile phase, and detected coulometrically in a screening-oxidation mode at +0.10 V and +0.35 V, respectively. The method was applied to the assay of in vitro enzymic O-methylation of catechol oestrogens. After 2- or 4-hydroxyoestradiol had been incubated with rat red blood cells in the presence of S-adenosylmethionine, the resulting guaiacols and unchanged substrate were percolated through an Extrelut-3 cartridge. The dried eluate was redissolved and directly injected. This simple procedure was as sensitive as the previously reported method using gas chromatography-mass spectrometry in a selected ion monitoring mode. PMID- 8408388 TI - Rapid and highly sensitive high-performance liquid chromatographic method for the determination of histamine and 3-methylhistamine in biological samples using fluorescamine as the derivatizing agent. AB - A highly sensitive and rapid high-performance liquid chromatographic assay for the determination of histamine and 3-methylhistamine in biological samples using 1-methylhistamine as the internal standard is described. Samples were purified and concentrated on cation-exchange columns and derivatized with fluorescamine. The lower detection limit was 20 pg on-column. Linearity was demonstrated up to 20 ng on-column. The samples could be derivatized simultaneously before injection and were stable for 7 days. The method was used for the determination of histamine and related compounds in coronary perfusates, extracts of homogenized rat hearts, and supernatants of stimulated peritoneal mast cells. PMID- 8408389 TI - Measurement of allantoin in urine and plasma by high-performance liquid chromatography with pre-column derivatization. AB - A method is reported for determination of allantoin in urine and plasma based on high-performance liquid chromatography (HPLC) and pre-column derivatization. In the derivatization procedure, allantoin is converted to glyoxylic acid which forms a hydrazone with 2,4-dinitrophenylhydrazine. The hydrazone appears as syn and anti isomers at a constant ratio. These derivatives are separated by HPLC using a reversed-phase C18 column from hydrazones of other keto acids possibly present in urine and plasma and then monitored at 360 nm. All components were completely resolved in 15 min. Both the reagents and derivatization products are stable. Recovery of allantoin added to urine and plasma was 95 +/- 3.7% (n = 45) and 100 +/- 7.5% (n = 64), respectively. The lowest allantoin concentration that gave a reproducible integration was 5 mumol/l. The between-assay and within-day coefficients of variation were 2.8 and 0.6%, respectively. PMID- 8408390 TI - Analysis of 6R- and 6S-tetrahydrobiopterin and other pterins by reversed-phase ion-pair liquid-chromatography with fluorimetric detection by post-column sodium nitrite oxidation. AB - A rapid and sensitive reversed-phase ion-pair liquid chromatographic system with fluorimetric detection by post-column sodium nitrite oxidation was established for measuring six pterin compounds (6R-5,6,7,8-tetrahydrobiopterin, 6S,5,6,7,8 tetrahydrobiopterin, 7,8-dihydrobiopterin, biopterin, pterin and D-neopterin). The coefficients of variation for these pterins were 0.705-3.714%, and the minimum detectable amount was ca. 10-20 pg at a signal-to-noise ratio of 3. A linear detector response was also verified. The concentrations of the pterin compounds in rat tissues were measured by the described method. Furthermore, by means of brain microdialysis, the output of pterin compounds from rat striatum was detected. Therefore, these results demonstrate that this system can be applied to analyses not only of various rat tissues but also of dialysates collected in vivo. PMID- 8408391 TI - New simplified procedures for the extraction and simultaneous high-performance liquid chromatographic analysis of retinol, tocopherols and carotenoids in human serum. AB - A short, simple extraction procedure and a sensitive reversed-phase high performance liquid chromatographic assay, which utilizes isocratic elution and detection either by a photodiode-array detector or by two detectors set at 300 and 450 nm, have been developed to measure retinol, tocopherols and several carotenoids in human serum simultaneously. By relying on characteristic UV visible spectra, seventeen carotenoids, retinol and alpha- and gamma-tocopherols were identified in concentrated serum extracts by use of the three-dimensional data mode of a photodiode-array detector. The presence of some recently reported carotenoids in human serum has been confirmed. PMID- 8408392 TI - Gas chromatographic-mass spectrometric quantitation of urinary 11-nor-delta 9 tetrahydrocannabinol-9-carboxylic acid after derivatization by direct extractive alkylation. AB - A gas chromatographic-mass spectrometric procedure for the quantitation of urinary 11-nor-delta 9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) has been developed in which the THC-COOH was derivatized to its corresponding methyl ester methyl ether derivative by direct extractive alkylation using tetrahexylammonium (THA+) counter-ion and iodomethane dissolved in toluene. The procedure utilised a sample volume of 2 ml and gave a detection limit of 0.25 ng/ml. The inter-run and intra-run coefficients of variation were 7.0% and 4.8%, respectively. The inter day standard curves were linear in the concentration range 0-300 ng/ml with a mean r = 0.9997 (n = 4). PMID- 8408393 TI - Determination of the enantiomers of zopiclone and its two chiral metabolites in urine using an automated coupled achiral-chiral chromatographic system. AB - The enantiomers of zopiclone and its two chiral N-desmethyl and N-oxide metabolites were determined in urine using a coupled achiral-chiral liquid chromatographic method. After liquid-liquid extraction, zopiclone and its two metabolites were quantified on a cyanopropyl column. After fluorimetric detection on the achiral system, the eluent was switched through a silica precolumn in order to trap and concentrate the analytes. Each fraction was then backflushed separately onto a carbamate cellulose chiral stationary phase in order to determine the enantiomeric ratios. The coupled system was automated with an autosampler and a switching valve programmed by an integrator. The method was validated, and a first trial was performed on urine samples of a volunteer treated with 15 mg of racemic zopiclone. PMID- 8408394 TI - Determination of the R,R- and S,S-enantiomers of vamicamide in human serum and urine by high-performance liquid chromatography on a Chiral-AGP column. AB - An enantioselective liquid chromatographic assay for the determinations of the R,R- and S,S-enantiomers of vamicamide, a potent anticholinergic drug, in human serum and urine is described. Racemic vamicamide and internal standard were purified from biological fluids using a two-step extraction procedure involving diethyl ether and 0.1% phosphoric acid. The overall recoveries of racemic vamicamide and internal standard were greater than 80%. The purified samples were measured by high-performance liquid chromatography on a Chiral-AGP column with ultraviolet absorbance detection at 260 nm. The standard curves for the analytes were linear from 10 to 200 ng/ml in serum and from 0.25 to 50 micrograms/ml in urine. The quantification limit of both enantiomers was 10 ng/ml for serum and 250 ng/ml for urine. Both intra-day and inter-day accuracy and precision data showed good reproducibility of the method. The assay has been applied for the analysis of vamicamide enantiomers in serum and urine samples from a healthy volunteer. PMID- 8408395 TI - High-performance liquid chromatographic method for simultaneous determination of clodronate and some clodronate esters. AB - A quaternary alkylmethylamine-bonded stationary phase has been used for the liquid chromatographic resolution of bisphosphonates. Clodronate and three of its esters were separated by this technique. Nitric acid (30 mM) was used as the mobile phase. The effect of the pH of the mobile phase on the retention, resolution, capacity factor and theoretical plates of the column was examined. Thorium-ethylenediaminetetraacetic acid-xylenol orange mixed ligand complex was used as a postcolumn reagent for bisphosphonates. Bisphosphonates react quantitatively with this complex in slightly acidic solutions, and a change in the absorbance of postcolumn reagent is used as a measure of the bisphosphonate concentration. The relative standard deviation (R.S.D.) values for samples in aqueous solutions were in the range 2.3-15.5% (area) and 1.7-5.9% (height). The detection limits for different compounds, Cmin, varied from 0.3 to 1.4 mg/l (area) and from 0.3 to 0.5 mg/l (height). In urine samples the R.S.D. (%) varied from 3.1 to 18.9 (area) and from 1.1 to 6.3 (height). The linear dynamic range was from the detection limit up to 16 mg/l. PMID- 8408396 TI - Gas chromatographic whole-cell fatty acid analysis as an aid for the identification of mixed mycobacterial cultures. AB - Gas chromatographic analysis of whole-cell fatty acids, secondary alcohols and mycolic acid cleavage products could be a useful technique in checking mixed mycobacterial cultures. The mixed cultures were confirmed when species-specific compounds of different mycobacterial species were detected in the same chromatogram. PMID- 8408397 TI - Plasma catecholamine levels in children. AB - While blood norepinephrine and epinephrine levels are well established in adults, literature data concerning concentrations in young children are conflicting. This situation prompted us to assay plasma catecholamines in 86 healthy subjects aged two days to fifteen years using a sensitive high-performance liquid chromatography technique with electrochemical detection. Norepinephrine and epinephrine concentrations remained elevated up until 24 months of age, then dropped progressively to adult levels between 24 and 36 months, with fluctuations. These fluctuations, which reflect the sensitivity of children to environmental factors, prevented demonstration of any catecholaminergic secretory abnormality by blood assays in subjects under two years of age. Urinary assays, for which established values exist, are the only applicable procedure in such patients. PMID- 8408398 TI - Simultaneous determination of retinol, alpha-tocopherol and retinyl palmitate in plasma of premature newborns by reversed-phase high-performance liquid chromatography. AB - A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of retinol, alpha-tocopherol and retinyl palmitate in plasma. Plasma containing an internal standard (tocol) was deproteinized with ethanol, then extracted with n-hexane. The organic layer was removed and evaporated under a nitrogen stream, and chromatographed on a reversed-phase RP-18 column using a water/acetonitrile-ethyl acetate/2-propanol gradient solvent system over 15 min at 305 nm. The recovery exceeded 93%. The detection limit was 0.1 microgram/ml for retinol, 1.3 micrograms/ml for alpha-tocopherol and 0.95 micrograms/ml for retinyl palmitate. The reproducibility, precision (expressed as coefficients of variation) and accuracy were less than 8% for all analytes. The small sample requirement, the simplicity of extraction, the short run-time and the good reproducibility make this procedure particularly useful for monitoring retinol and alpha-tocopherol supplementation in premature newborns. PMID- 8408399 TI - Quantification of dimethindene in plasma by gas chromatography-mass fragmentography using ammonia chemical ionization. AB - A gas chromatographic-mass fragmentographic method using ammonia chemical ionization for the determination of dimethindene in human plasma is described. The drug was isolated from plasma by liquid-liquid extraction with hexane-2 methylbutanol. Plasma components were separated on a capillary column coated with chemically bonded methyl silicone. For detection of dimethindene, its quasi molecular ion (M + H+) was mass fragmentographically monitored after chemical ionization with ammonia as reagent gas. Dimethindene was quantified using methaqualone as the internal standard: the quantification limit in plasma was 0.2 ng/ml, the within-run precision was 8.0% and the inter-run precision 5.6%. The plasma concentration-time profile was established after a single dose of 4 mg of dimethindene with an average maximum concentration of 5.5 ng/ml, detectable up to 48 h post application. PMID- 8408400 TI - Investigation of the human metabolism of antipyrine using coupled liquid chromatography and nuclear magnetic resonance spectroscopy of urine. AB - The potential of coupled high-performance liquid chromatography-nuclear magnetic resonance spectroscopy for the detection and identification of drug metabolites has been investigated by direct analysis of human urine collected following administration of antipyrine. This approach provided a rapid method of characterizing the major human urinary metabolites of this drug and promises to be of widespread value in structural studies of xenobiotic metabolites. PMID- 8408401 TI - Determination of ciprofloxacin levels in chinchilla middle ear effusion and plasma by high-performance liquid chromatography with fluorescence detection. AB - An isocratic high-performance liquid chromatographic method has been developed to determine ciprofloxacin levels in chinchilla plasma and middle ear fluid. Ciprofloxacin and the internal standard, difloxacin, were separated on a Keystone ODS column (100 x 2.1 mm I.D., 5 microns Hypersil) using a mobile phase of 30 mM phosphate buffer (pH 3), 20 mM triethylamine, 20 mM sodium dodecyl sulphate acetonitrile (60:40, v/v). The retention times were 3.0 min for ciprofloxacin and 5.2 min for difloxacin. This fast, efficient protein precipitation procedure together with fluorescence detection allows a quantification limit of 25 ng/ml with a 50 microliters sample size. The detection limit is 5 ng/ml with a signal to-noise ratio of 5:1. Recoveries (mean +/- S.D., n = 5) at 100 ng/ml in plasma and middle ear fluid were 89.4 +/- 1.2% and 91.4 +/- 1.6%, respectively. The method was evaluated with biological samples taken from chinchillas with middle ear infections after administering ciprofloxacin. PMID- 8408402 TI - Determination of amoxicillin in human plasma by high-performance liquid chromatography and solid phase extraction. AB - A high-performance liquid chromatographic method using solid phase extraction has been developed for the determination of amoxicillin in human plasma. After concentration of amoxicillin on a C8 cartridge, the cartridge was eluted on-line to a reversed-phase column packed with 5 microns Chromspher C18. The mobile phase was methanol-0.08 M phosphate buffer (pH 7.6) (20:80), which contained 0.01 M tetrabutylammonium dihydrogenphosphate. The column effluent was monitored by UV spectrophotometry at 234 nm. The method has proved to be reliable and is used in bioavailability studies for the development of new amoxicillin formulations. PMID- 8408403 TI - Automated high-performance liquid chromatographic method for determination of furosemide in dog plasma. AB - An automated high-performance liquid chromatographic method for the determination of the diuretic drug furosemide has been established. Dog plasma was injected directly into a two-column system with a BSA-ODS (ODS column coated with bovine serum albumin) precolumn and a C18 analytical column for the separation of furosemide. The two columns were automatically switched. Furosemide remained trapped on the precolumn while proteins were eluted to waste. After column switching, furosemide was washed onto the analytical column and analysed without interference. The greatest advantage of the method is its easy performance without manual sample preparation; it requires no extraction or deproteinization. The method allows determination of 0.1-10 micrograms/ml of furosemide with accuracy and precision comparable with previously reported values. The coefficients of variation obtained from replicate measurements of 1 microgram/ml and 5 micrograms/ml samples were 1.65% and 2.40%, respectively. This method was used to measure the plasma levels of furosemide in beagle dogs to whom the drugs was administered, as a reference, in a toxicological study. PMID- 8408404 TI - Studies on a new cross-axis coil planet centrifuge for performing counter-current chromatography. I. Design of the apparatus, retention of the stationary phase, and efficiency in the separation of proteins with polymer phase systems. AB - An improved model of the cross-axis synchronous flow-through coil planet centrifuge has been designed in light of previous studies. The apparatus has a versatile feature in that both analytical and preparative columns can be accommodated in both off-center and central positions. Each has merit in separations. Retention of stationary phase was examined with various two-phase solvent systems used for the separation of biopolymers. Both analytical and preparative columns showed satisfactory retention of the stationary phase under optimum conditions. The apparatus was evaluated in separation of a set of protein samples using a polyethylene glycol-potassium phosphate biphasic system. In both types of columns all proteins were resolved with partition efficiencies of 260 to 670 theoretical plates. Further studies indicated that the relatively low partition efficiency of proteins is mainly attributed to their high molecular mass or molecular heterogeneity within each species rather than due to the high viscosity of the polymer phase system. PMID- 8408405 TI - Studies on a new cross-axis coil planet centrifuge for performing counter-current chromatography. II. Path and acceleration of coils and comparison with type J coil planet centrifuge. AB - A comprehensive approach is introduced using parametric equations to describe the motion induced by type J and cross-axis type coil planet centrifuges. The plots of a line parallel to the coil axis are given. The centrifugal forces are then analyzed in two ways. Three-dimensional graphs show their geometry and the relative intensity of the lateral component is compared to that of the perpendicular component. This allowed a better understanding of the differences in paths and acceleration fields induced by the two types of counter-current chromatography devices. PMID- 8408406 TI - Studies on new cross-axis coil planet centrifuge for performing counter-current chromatography. III. Speculations on the hydrodynamic mechanism in stationary phase retention. AB - Retention of the stationary phases of one organic-aqueous solvent system and three aqueous-aqueous polymer solvent systems was investigated on a cross-axis coil planet centrifuge. A graphic statistical treatment of all the data highlighted the prevailing effect of the inward-outward elution mode. A simplified model was proposed and studies on the paths and accelerations of cross axis devices described in the previous paper provided explanations about the observed hydrodynamic behaviors. PMID- 8408407 TI - Observation of a conformational effect in peptide molecule by reversed-phase high performance liquid chromatography. AB - RP-HPLC of analogues of oxytocin containing a reduced peptide bond was studied using various columns and buffers. Anomalous behavior of one analogue served as a basis for the discussion of possible conformational consequences of this substitution. PMID- 8408408 TI - Analysis of paralytic shellfish poisoning toxins by automated pre-column oxidation and microcolumn liquid chromatography with fluorescence detection. AB - Periodate oxidation of the toxins responsible for paralytic shellfish poisoning (PSP) yields fluorescent purines suitable for trace analysis by reversed-phase LC. Mobile phases containing perfluorinated acids, such as heptafluorobutyric acid, as ion-pair agents were found to provide high capacity factors for the oxidized products. Gradient elution on a microbore column with large volume injections and fluorescence detection permitted the detection of femtomole quantities of PSP toxins. A fully automated pre-column oxidation procedure was developed for an LC autosampler system in order to improve precision and allow unattended analyses. The complete method was applied successfully to various samples, including shellfish and toxic phytoplankton. PMID- 8408409 TI - Determination of ionic species formed during growth of Escherichia coli by capillary isotachophoresis. AB - The ionic species that are formed during the microbial growth of Escherichia coli were determined by capillary isotachophoresis as a function of the time of cultivation. This formation was indicated by the change in a sum parameter, the impedance of the nutrient broth, measured by a special electrode system. Based on the determination of the individual ions formed under the given conditions (identified as acetate, lactate, alpha-ketoglutarate, fumarate, ammonium and probably a simple amine), the change in conductivity was calculated and compared with that obtained by the impedance measurement of the bulk medium. From the results it can be concluded that the change in the sum parameter as a function of time is originated by the ions determined. PMID- 8408410 TI - Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography. AB - The peculiarities of zone migration and band broadening in the reversed-phase gradient HPLC of proteins were investigated. In the isocratic mode a critical composition of the mobile phase was found at which all proteins regardless of their molecular mass migrate with equal velocity and have a capacity factor equal to the phase ratio (VP/V0), i.e., the same capacity factor as a marker of total accessible volume would have in steric exclusion chromatography. It is shown that steric exclusion conditions are never achieved in gradient HPLC. In the first (adsorption stage) of gradient elution where the separation takes place the velocity of a protein increases until it becomes equal to the velocity of the desorbing solvent front at a critical distance X0 from column entrance. Strong broadening is characteristic of this stage. In the second (critical) stage the protein travels the remaining distance (L-X0) with the velocity of the solvent. A definition of X0 is given allowing one very simple calculation of the minimum permissible column length as a function of gradient steepness, mobile phase velocity and protein adsorption parameter. When x = X0 the protein zone has the smallest dispersion. Making L < X0 is especially disadvantageous, as it leads to anomalous bandspreading. The theory of gradient HPLC was refined on this basis and the usefulness of this approach in high-performance membrane chromatography is demonstrated. PMID- 8408411 TI - High-performance gel-permeation chromatographic analysis of protein aggregation. Application to bovine carbonic anhydrase. AB - Protein association in bovine carbonic anhydrase (bCA) was investigated by high performance gel-permeation chromatography (HPGPC). It was shown that bCA undergoes a time-dependent aggregation after freezing and storing at -20 degrees C. The addition of 50% ethylene glycol or glycerol prevented the aggregation. The HPGPC data were confirmed by the use of cross-linking with subsequent sodium dodecyl sulphate-polyacrylamide gel electrophoretic analysis. The esterase activity of the aggregated bCA was comparable to that of the monomer, suggesting that the observed protein association does not affect the esterase catalytic centre of bCA. This study illustrates how HPGPC can be a convenient and reliable tool for monitoring storage-induced aggregation of proteins. PMID- 8408412 TI - Characterization of vitamin D3 metabolites using continuous-flow fast atom bombardment tandem mass spectrometry and high-performance liquid chromatography. AB - A mass spectrometric method for the detection of vitamin D3 metabolites is described. This method involves the derivatization of the metabolites by cycloaddition with 4-phenyl-1,2,4-triazoline-3,5-dione, followed by their characterization by continuous-flow fast atom bombardment (CF-FAB) tandem mass spectrometry (MS-MS) and high-performance liquid chromatography (HPLC). Using HPLC, this derivatization has been shown to increase the UV detectability of 25 hydroxyvitamin D3 by about 5-fold. The FAB spectra of the adducts are dominated by peaks corresponding to a protonated molecule and a fragment ion derived in part from the loss of the side chain. Multiple reaction monitoring (MRM) of this transition by MS-MS may be utilized for trace level analysis of vitamin D metabolites. Sample introduction by flow injection yields detection limits in the low nanogram to high picogram range, whereas the use of on-line capillary LC has been found to decrease the detection limits to the low picogram level. PMID- 8408413 TI - Determination of pesticides in drinking water by on-line solid-phase disk extraction followed by various liquid chromatographic systems. AB - C-18 Empore extraction disks were coupled on-line with liquid chromatography rapid scanning UV-VIS detection and post-column fluorescence detection for the isolation and trace enrichment of various pesticides [carbamates, (aldicarb, carbofuran, carbaryl), carbamate transformation products (TPs) (aldicarb sulfoxide, aldicarb sulfone, 3-hydroxycarbofuran, 3-hydroxy-7-phenol carbofuran, 3-keto-carbofuranphenol and 3-ketocarbofuran) and herbicides (chlortoluron, isoproturon and metolachlor)] spiked at concentration levels of 0.2 and 5 micrograms/l in drinking water samples. Recoveries were dependent on the pesticide level and preconcentrated water volume (50 ml to 1000 ml) using LC with rapid scanning UV-VIS detection. The same on-line system coupled with LC-post column derivatization fluorescence detection has needed only 10 ml of water to achieve similar levels of determination for the carbamate insecticides. PMID- 8408414 TI - Quantitation of 5-methylcytosine by one-dimensional high-performance thin-layer chromatography. AB - A method for the quantitative analysis of DNA 5-methylcytosine by one-dimensional high-performance thin-layer chromatography using alkylamino modified silica (HPTLC-NH2) plates is described. The preparative method is simple, involving enzymatic digestion of DNA with micrococcal nuclease and phosphodiesterase II to 3'-monophosphate nucleosides, conversion by T4 polynucleotide kinase to 32P labeled 3',5'-bisphosphate nucleosides, and chromatographic separation of nuclease P1-cleaved 5'-monophosphate nucleosides. The weak, basic anion exchanger property of the HPTLC-NH2 plate enables separation of multiple samples in one dimension, whereas traditional polyethyleneimine cellulose plates require development of individual samples in two dimensions for analysis of 5' methylcytosine. PMID- 8408415 TI - Determination of total vitamin C in fruits by capillary zone electrophoresis. AB - A simple capillary zone electrophoretic (CZE) method is described for the rapid determination of ascorbic acid and dehydroascorbic acid, the physiologically active forms of vitamin C, in fruits. The electrophoretic run was accomplished in 9 min on a coated capillary column using 20 mM phosphate buffer (pH 7.0). Total ascorbic acid was determined by first reducing the dehydroascorbic acid to ascorbic acid by treatment with DL-homocysteine. This reaction was complete in 15 min and total ascorbic acid determination was performed immediately. The data obtained by CZE were in good agreement with HPLC data. PMID- 8408416 TI - Displacement chromatography on cyclodextrin silicas. IV. Separation of the enantiomers of ibuprofen. AB - A displacement chromatographic method has been developed for the preparative separation of the enantiomers of ibuprofen using a beta-cyclodextrin silica stationary phase. The retention behavior of ibuprofen was studied in detail: the log k' vs. polar organic modifier concentration, the log k' vs. pH, the log k' vs. buffer concentration and the log k' vs. 1/T relationships; also, the alpha vs. polar organic modifier concentration, the alpha vs. pH, the alpha vs. buffer concentration and the log alpha vs. 1/T relationships have been determined in order to find the carrier solution composition which results in maximum chiral selectivity and sufficient, but not excessive solute retention (1 < k' < 30). 4 tert.-Butylcyclohexanol, a structurally similar but more retained compound than ibuprofen, was selected as displacer for the separation. Even with an alpha value as small as 1.08, good preparative chiral separations were observed both in the displacement mode and in the overloaded elution mode, up to a sample load of 0.5 mg. PMID- 8408417 TI - Characterization of the binding and chiral separation of D- and L-tryptophan on a high-performance immobilized human serum albumin column. AB - High-performance affinity chromatography was used to study the separation and binding of D- and L-tryptophan on an immobilized human serum albumin (HSA) column. Frontal analysis and zonal elution studies indicated that both D- and L tryptophan were binding to single but distinct sites on HSA. L-Tryptophan bound to the indole site of HSA. D-Tryptophan had indirect interactions with the warfarin site of HSA but no interactions with the indole site. The association constants for the binding of D- and L-tryptophan at pH 7.4 and 25 degrees C were 0.4 x 10(4) and 2.7 x 10(4) M-1, respectively. The value of delta G for these sites ranged from -5.2 to -5.7 kcal/mol (1 cal = 4.184 J) and had a significant entropy component. The effects of varying the pH, phosphate concentration, temperature and polarity of the mobile phase on the binding of D- and L tryptophan to HSA were examined. The role of sample size in determining peak shape and retention was also considered. From these data, general guidelines were developed for the separation of D- and L-tryptophan on immobilized HSA. Under optimized conditions the enantiomers were separated in less than 2 min with baseline resolution. PMID- 8408418 TI - Capillary zone electrophoresis of malto-oligosaccharides derivatized with 8 aminonaphthalene-1,3,6-trisulfonic acid. AB - Malto-oligosaccharides were derivatized via their reducing end with 8 aminonaphthalene-1,3,6-trisulfonic acid by reductive amination, and the separation and electrophoretic migration behavior of the labelled sugars were investigated by capillary zone electrophoresis. Series of linear malto oligosaccharides were found particularly suitable for both the study of the effect of the operating conditions on the separation and the investigation of the relationship between the electrophoretic mobility and the molecular size of the homologues. The electrophoretic mobility of the malto-oligosaccharide conjugates was found to be a linear function of the molecular mass to the negative two thirds power. The sugar derivatives employed here carry three negative charges due to the presence of the dissociated sulfonic acid groups even at strongly acidic pH. Therefore, the analytes can migrate in the electric field without interference by electroendosmotic flow and/or wall adsorption in uncoated silica capillaries at low pH. As a result, the separation of these carbohydrate conjugates can be carried out under such conditions with high speed and efficiency in free solution, i.e., without an anticonvective medium such as a gel or a viscous polymer solution. Appropriate use of triethylammonium phosphate buffer, pH 2.5, as the background electrolyte improves not only the reproducibility, but also the efficiency and speed of the separation. The labelled sugars allow monitoring of the separation by UV detector or laser induced fluorescence detector with concomitant enhancement of analytical sensitivity. PMID- 8408419 TI - Separation of isoforms of Serratia marcescens nuclease by capillary electrophoresis. AB - Three S. marcescens nuclease isoforms differing mainly in charge (native nuclease with pI 6.8 and two minor isoforms with pI 7.3 and 7.4) were separated using several different modes of high-performance capillary electrophoresis. Separation of the isoforms by free solution capillary electrophoresis was unsatisfactory. Separation by micellar electrokinetic capillary chromatography was therefore investigated in detail and the method optimized with respect to pH and sodium dodecyl sulphate concentration; in addition, the effect of adding various substances to control dispersion and avoid analyte adsorption at the capillary wall was examined. Under optimal conditions there was almost complete baseline separation of the two isoforms with basic pI whereas there was only partial separation of the native form and the isoform with pI 7.4. With capillary isoelectric focusing there was complete baseline separation of the native nuclease and the other two isoforms. PMID- 8408420 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXXI. O-phosphoserine as a new chelating ligand for use with hard Lewis metal ions in the immobilized-metal affinity chromatography of proteins. AB - Conditions for the immobilization of O-phosphoserine (OPS) to epoxy-activated Sepharose CL-4B are described. The binding behaviour of OPS and iminodiacetic acid (IDA) immobilized onto Sepharose CL-4B, toward the hard Lewis metal ions Al3+, Fe3+, Ca2+ and Yb3+, and Cu2+ ion as a borderline metal ion control, over the pH range pH 4.0 to pH 8.0, was examined. Immobilized OPS shows a stronger affinity for Fe3+ and Al3+ ions but a lower affinity for Cu2+ and Yb3+ ions, compared to immobilized iminodiacetic acid (IDA), over the equilibrating range examined. Immobilized OPS-Mn+ was screened for protein binding using as model proteins tuna heart cytochrome c (THCC), horse myoglobin (HMYO) and hen egg while lysozyme (HEWL) over the pH range 5.5 to 8.0. Immobilized OPS-Fe3+ bound THCC under all the examined equilibrating conditions, bound HMYO between pH 5.5 and pH 7.0 and did not bind HEWL under any condition examined. Immobilized OPS thus presents an additional mode of metal ion and protein selectivity in immobilized metal affinity chromatography. PMID- 8408421 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXIX. Ceramic-based particles as chemically stable chromatographic supports. AB - Porous zirconia based particles have been modified using different derivatisation procedures. The modified particles were characterised in terms of their accessible surface areas and degree of surface coverage of the bounded or physicoated phases utilising the strong and specific adsorption of phosphate ions to the zirconia surface. The hydroxyl group density was determined by a 1H NMR technique. The particles were modified by immobilising different silanes to introduce either hydrophobic ligands or reactive groups onto the zirconia surface. In the latter case, various ligands were then covalently attached to the activated supports. Using this type of modification, n-octadecyl- (C18), carbohydrate- and Cibacron Blue F3GA-modified zirconia particles were produced. Furthermore, polymeric coated particles were prepared either by using polybutadiene or by cross-linking the carbohydrate modified sorbents. The pH stability of the different sorbents were determined in batch experiments and under chromatographic conditions. The leakage of ligands was monitored by UV absorption and by employing radioactively labelled ligands. The performance of the C18 reversed-phase modified zirconia in packed columns was also used as an indicator of changes in the surface chemistry following pH stability tests. The experimental results indicate that the Cibacron Blue F3GA dye-modified sorbent was stable up to pH 10.5, the C18 reversed-phase packing up to pH 13 and the carbohydrate-bonded phase up to pH 12. These investigations substantiate the favourable chemical and physical characteristics anticipated for surface modified zirconias for potential use as chromatographic adsorbents. PMID- 8408422 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXX. Modified porous zirconia as sorbents in affinity chromatography. AB - The utilisation of organosilanes to introduce active chemical groups onto zirconia surfaces, suitable for the subsequent immobilisation of proteins or other biomimetic ligands, is described. Two different types of porous zirconia based particles with nominal pore diameters of 160 and 1000 A pore size were modified with two different affinity ligands. In the first case, methods to immobilise iminodiacetic acid-Cu(II) and its application in Cu(II) immobilised metal ion affinity chromatography (IMAC) were established. In the second series of experiments, concanavalin A was immobilised and the interaction of this lectin with the enzyme horseradish peroxidase examined. For both systems, adsorption isotherms were recorded as batch experiments. In each case, the experimental results could be fitted to langmuirean type adsorption isotherms, indicating that under the chosen conditions only one type of interaction is present, with nonspecific interactions with the support surface playing an insignificant role. These studies document the potential of surface modified zirconia particles for the immobilisation of chemical ligands or proteins for use in biospecific affinity chromatography and immobilised enzyme bioreactors. PMID- 8408423 TI - Separation of membrane-embedded tryptic peptides of Na,K-ATPase by size-exclusion chromatography. AB - Several attempts to separate hydrophobic tryptic and cyanogen bromide-digested short peptides from Na,K-ATPase, using HPLC and different acid-organic solvent gradients, failed because of the insolubility of the peptides in the initial or final solvents of the gradients used for elution. Therefore, we opted to use a detergent-containing mobile phase. For sodium dodecyl sulphate-solubilized tryptic peptides of M(r) 7 x 10(3)-100 x 10(3), elution on a TSK-G3000SW size exclusion column successfully separates families of peptides with a resolution of M(r) 5 x 10(3)-10 x 10(3). Peptides in these size ranges can then be resolved completely by tricine-sodium dodecyl sulphate gel electrophoresis, and identified by microsequencing after transfer to polyvinylidene difluoride paper. For separation of smaller peptides a Biosep-SEC-S2000 column, eluted at slow flow rates, was evaluated. Use of ammonium chloride buffer allows sensitive detection at 214 nm. The separated fractions are resolved and identified on 16.5% tricine gels. Reasonable resolution has been obtained with defined cyanogen bromide fragments of myoglobin. Resolution of small tryptic and cyanogen bromide fragments of Na,K-ATPase is less successful, but the experiments suggest ways of improving the resolution of peptides in the range M(r) 2 x 10(3)-10 x 10(3). PMID- 8408424 TI - Rapid high-sensitivity peptide mapping by liquid chromatography-mass spectrometry. AB - Toward a complete LC-MS mapping system for peptides and proteins, we have coupled a precision-flow microbore HPLC system to an electrospray single quadrupole mass spectrometer. The HPLC system allows fast separation of protein digests with UV detection at the low pmol level. A 2 microliters/min portion (1:25) of the effluent is passed into a high-sensitivity electrospray MS system. The electrospray source allows for molecular mass associated ions (MH+, MH2(2+), MH3(3+), etc.) to be generated as well as collision induced dissociation of these ions before MS analysis. After LC-MS runs, with or without partial fragmentation, the data generated are largely interpreted by identification of predicted peptides, incompletely digested peptides, unusual peptide cleavages, and so on, using appropriate integrated software (PEPMAP, PEPMATCH). Examples of peptide mapping at the low pmol level using this integrated system will be shown (e.g., of the protein human growth hormone and of the glycoprotein, tissue plasminogen activator). PMID- 8408425 TI - Effect of the alpha-amino group on peptide retention behaviour in reversed-phase chromatography. Determination of the pK(a) values of the alpha-amino group of 19 different N-terminal amino acid residues. AB - We have examined the contribution of the alpha-amino group to retention behaviour for peptides in reversed-phase chromatography using two series of peptide analogues, one containing an N alpha-acetylated terminal and the other containing an alpha-amino group (non-acetylated). The effect of the alpha-amino group, at pH 2, on the hydrophobicity of the side-chain of the N-terminal residue was obtained by referencing the retention time of the acetylated or non-acetylated peptide to the retention time of a glycine analogue. It was shown that the presence of an alpha-amino group could decrease or increase the hydrophobicity of the side-chain of the N-terminal residue with respect to the hydrophobicity of the side-chain in the absence of an alpha-amino group. The effect was also shown to be sequence dependent, with respect to the N-terminal residue. Increasing pH was shown to increase retention time dramatically for the non-acetylated analogues, through the deprotonation of the alpha-amino group. By separating pairs of acetylated/non acetylated analogues over the pH range 2-9, it was possible to determine the pK(a) of the alpha-amino group, where it was shown that the pK(a) was dependent on two probable factors: (1) the inherent hydrophobicity of the stationary phase; and (2) the amino acid substituted in the N-terminal position. Interestingly, the pK(a) values determined were very similar to that found in proteins. It was also possible to determine the pK(a) values of some of the substituted amino acids containing ionizable side-chains. This study shows that, in order to understand fully the retention behaviour of peptides containing an alpha-amino group in reversed-phase chromatography, one must incorporate an alpha-amino group contribution and its effect on the hydrophobicity of the side-chain of the N terminal residue. PMID- 8408426 TI - Analytical methods for differentiating minor sequence variations in related peptides. AB - A proline-rich peptide was isolated and purified to homogeneity from an extract of bovine neutrophil granules using semi-preparative RP-HPLC. The relative molecular mass of the peptide (called Bac-X) was determined by ionspray MS to be 5149 +/- 0.5. The amino acid composition of the peptide was characterized by its limited number of amino acid types, which included a high proline (43.3%) and arginine content (20.3%), and hydrophobic residues. Bac-X had similar characteristics to Bac-5, a previously characterised bactenecin of bovine neutrophil granules, with respect to its proline, arginine and hydrophobic amino acid content, molecular mass and antibacterial specificity. Tryptic and N bromosuccinimide digestion of Bac-X produced fragments with masses (M(r) 785 and 4224 and 3100 respectively) consistent with those expected from a peptide with the reported sequence of Bac-5. Bac-X differed from Bac-5 in the number of amino acid residues (43 for Bac-X versus 42 for Bac-5) and contained glycine which Bac 5 did not. However, the calculated molecular mass of the peptide, based on the amino acid compositional data, did not match the experimental value. The purified peptide could not be sequenced by Edman degradation due to apparent blockage of the N-terminus. Partial sequence information, obtained by LC-MS and collision induced dissociation MS-MS analysis of a M(r) 785 tryptic fragment of Bac-X, showed that this peptide contained a six residue sequence (-RFPPIR-) not found in Bac-5 which, based on its reported sequence, contained a M(r) 785 tryptic fragment with the sequence -FRPPIR-. This difference in sequence of Bac-X compared with Bac-5 illustrates the application of electrospray (ionspray) MS techniques to the detection and identification of minor differences in related protein/peptide forms. PMID- 8408427 TI - Characterisation of TNF-alpha-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography tandem mass spectrometry. AB - Ion-spray triple quadrupole mass spectrometry and high-performance liquid chromatography were used to investigate the products from the solid phase synthesis of (H)-Leu-Thr-Glu-Asn-(OH), a TNF-alpha active-site probe. The target sequence was assembled using tert.-butoxycarbonyl (Boc) chemistry in stepwise fashion from the C-terminal on an Boc-Asn-OCH2-Pam-copoly(styrene-divinylbenzene) resin [Pam = 4-(carboxamidomethyl)benzyl ester]. The crude product was deprotected and cleaved from the resin by HF-p-cresol treatment for 1 h at 0 degrees C. HPLC analysis at 214 nm indicated two late-eluting major products and an early-eluting product. Preparative HPLC demonstrated that the early-eluting product contained ca. 80% of the expected recovered sample mass. Each component was then directly analysed by mass spectrometry and tandem mass spectrometry. The early eluting peak was confirmed as the desired LTEN sequence. Synthesis of the same sequence using 9-fluorenyl methoxycarbonyl (Fmoc) chemistry gave an identical product and confirmed the above analysis. The most significant by product was derived from arylation of the glutamyl group by the quencher p cresol. The likely origins of the by-products are discussed. PMID- 8408428 TI - Isolation of carboxyl-termini and blocked amino-termini of viral proteins by high performance cation-exchange chromatography. AB - The strong cation-exchanger, PolySulfoethyl Aspartamide, has been assessed as a medium for isolation of carboxyl-terminal and blocked amino-terminal peptides from tryptic digests of small quantities of viral proteins. Peptides with a single positive charge, the blocked amino-terminal peptides of ovalbumin and the Newcastle disease virus (NDV) matrix protein and carboxyl-terminal peptides of ovalbumin and the NDV nucleocapsid protein, eluted in early ion-exchange fractions and were readily isolated in homogeneous form by subsequent reversed phase HPLC. Some early ion-exchange fractions also contained singly charged peptides derived by "chymotryptic-like" cleavage, whilst other peptides eluted in these fractions due to their highly acidic character. Terminal sequences with additional basic residues were isolated from later eluting ion-exchange fractions. Peptides with this property included the blocked amino-terminus of the NDV nucleocapsid protein and a portion of the carboxyl-terminus of the NDV matrix protein. Hitherto undescribed polymorphism in the amino-terminal region of ovalbumin was revealed in this study. Truncated peptides from the carboxyl terminus of the NDV matrix protein were also detected. The presence of these peptides could be a reflection of carboxyl-terminal processing of the matrix protein. The strategy described herein should be of general utility for selective microisolation of carboxyl-terminal peptides and blocked amino-terminal peptides from tryptic digests of proteins. PMID- 8408429 TI - Human pepsin 3b peptide map sequence analysis, genotype and hydrophobic nature. AB - Peptides from a Staphylococcus aureus (V8) proteinase digest of human pepsin 3b have been identified by amino acid sequence analysis. Only 137 out of 326 expected residues were detected from the C and N terminal regions of the molecule. Comparison with amino acid sequences derived from nucleotide analysis of three different pepsinogen A genes, identified 2 out of 4 possible substitutions. The presence of valine at position 30 and leucine at 291 indicates that the major pepsin component of gastric juice, pepsin 3b, corresponds to pepsinogen genotype PGA-3. Reversed-phase chromatography of native human pepsin 3b on C4 (300 A), C18 (300 A) or polymer (1000 A) columns was optimal on the C4 column and gradient elution with 2-propanol rather than acetonitrile. Denaturation of the protein in guanidinium hydrochloride, urea or high pH resulted in irreversible column retention. The marked hydrophobicity of denatured pepsin 3b may thus explain why the central segment of the protein was not revealed by peptide map analysis. PMID- 8408430 TI - Structural characterisation of native and recombinant forms of the neurotrophic cytokine MK. AB - The retinoic acid (RA)-inducible midkine (MK) gene encodes a heparin-binding protein which can induce neurite outgrowth in cultured mammalian embryonic brain cells. This cytokine shares 65% amino acid sequence identity with another RA inducible cytokine, pleiotropin (PTN). Both proteins contain 10 conserved cysteine residues, all of which appear to be disulphide linked. MK and PTN are also rich in lysine and arginine residues rendering them susceptible to proteolysis during purification, and making large-scale preparation of these molecules inherently difficult. Recombinant MK has been expressed as a fusion protein using a pGEX vector transfected into E. coli. To enable refolding of MK, the fusion protein was stored in solution at 4 degrees C for 14 days in the presence of dithiothreitol (DTT). Thrombin cleavage of the fusion protein, post storage, typically generated 5 mg of MK per litre of bacterial pellet. To establish the structural integrity of the recombinant product, we have analysed the refolding kinetics and compared the disulphide bond assignment of recombinant MK with that of native MK and native PTN. The synergistic use of micropreparative HPLC, to separate and recover in small eluant volumes enzymatically derived peptide fragments, with matrix assisted laser desorption mass spectrometry (MALD MS) and N-terminal sequence analysis has allowed the unambiguous identification of the disulphide bonded fragments of native and recombinant MK. The disulphide bond assignment of MK is C12-C36, C20-C45, C27-C49, C59-C91 and C69-C101, and is equivalent to that of PTN. PMID- 8408431 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXVIII. Effect of D-amino acid substitutions on the reversed-phase high performance liquid chromatography retention behaviour of neuropeptide Y[18-36] analogues. AB - The reversed-phase high-performance liquid chromatographic (RP-HPLC) gradient elution behaviour of a series of peptides related to Neuropeptide Y (NPY) has been investigated. The peptides studied included NPY, NPY[13-36], NPY[18-36] and a series of 16 analogues of NPY[18-36], each with a single D-amino acid substitution. Chromatographic parameters which relate to the interactive contact area and the binding affinity have been evaluated with two different stationary phase ligands and two organic modifiers. The results demonstrate that D-amino acid substitutions in the sequence region encompassing amino acid residues NPY[27 31] of these NPY[18-36] peptides significantly influence the interactive behaviour of these peptides relative to the unsubstituted NPY[18-36] molecule, while substitutions in the N- and C-terminal regions had little effect. Further, these results indicate that, in hydrophobic environments, NPY[18-36] adopts a significant degree of secondary structure which is severely disrupted by the presence of the D-amino acids in the central portion of the molecule. PMID- 8408432 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXVI. Modelling of protein adsorption with non-porous and porous particles in a finite bath. AB - Analytical solutions for a mathematical model describing dynamic adsorption processes of proteins onto non-porous adsorbent particles in a finite bath are presented. The model, based on the Langmuir adsorption isotherm, has been applied to experimental data obtained with affinity and ion-exchange adsorbents. The external film mass transfer resistance, as well as the rate of surface interaction between proteins and adsorbents, have been taken into account. The model has been extended to the case of adsorption onto porous particles by employing a linear driving force approximation for describing mass transfer in the pore fluid. This approach enables the derivation of an effective overall liquid phase mass transfer coefficient, permitting subsequent adaptation of the analytical solutions developed for non-porous particles. The evaluation of the effective liquid phase mass transfer coefficients is also described. Examples of a comparison between predicted and experimental dynamic adsorption curves for both dye-affinity and ion-exchange systems are presented. The application of the model for predicting the optimum operating conditions is discussed. PMID- 8408433 TI - High-performance liquid chromatography of amino acids, peptides and proteins. CXXXII. Optimisation of operating parameters for protein purification with chromatographic columns. AB - In large-scale chromatography, process optimisation is one of the key elements for success. This paper presents a method for determining the optimum operating parameters for affinity and ion-exchange chromatographic columns when used for protein purification. Based on a mathematical model developed as part of our association investigations, computer programs have been developed to describe the dynamic relationships acting within the chromatographic system. Two basic operating parameters, the flow-rate and the effluent concentration at which the adsorption stage is terminated, can be optimised to give a maximum production rate. The sample loading volume and the processing time then can be determined. The effect of washing conditions on the production rate and the yield is also discussed. Examples are given for a specific system where the optimisation is based on the yield and the percentage utilisation of the column capacity. Contour plots are generated to aid the determination of the range of controlling parameters, and to guide further system design. PMID- 8408434 TI - Use of a porous graphitised carbon column for the high-performance liquid chromatography of oligosaccharides, alditols and glycopeptides with subsequent mass spectrometry analysis. AB - HPLC using a porous graphitised carbon (PGC) column eluted in acetonitrile aqueous trifluoroacetic acid has been shown to give complementary chromatography to reversed-phase (ODS) HPLC for separation of peptides and glycopeptides. The PGC column can also be used for separation of oligosaccharides and oligosaccharide alditols released from protein by enzymes (N-linked chains) or base-borohydride degradation (O-linked chains). The advantages are that peptides, glycopeptides, reducing oligosaccharides, sialylated oligosaccharides and oligosaccharide alditols can be chromatographed under the same conditions. The samples can be readily recovered by evaporation for sensitive liquid secondary ion mass spectrometric (LSI-MS) analysis and there is no contamination or deterioration of chromatography from column leakage. LSI-MS analysis revealed that complete peak separation of all of the possible oligosaccharide components of the standard glycoproteins fetuin and bovine submaxillary mucin was not achieved. However, PGC remains as a useful adjunct to other HPLC profiling and separation techniques in particular where subsequent MS analysis is desired. PMID- 8408435 TI - Study of lectin-ganglioside interactions by high-performance liquid affinity chromatography. AB - A high-performance affinity column containing immobilized modified GM1 (lyso-GM1) was used to study the binding of an endogenous human brain lectin (HBL) in comparison with other carbohydrate-binding proteins. The proteins are previously converted into biotinylated derivatives. Detection of biotinylated proteins in the eluates by a microtitre plate assay ensures good sensitivity. The maximum binding capacity of the adsorbent for HBL is obtained in Tris buffer supplemented with beta-mercaptoethanol. The binding is inhibitable by specific sugar. It is concluded that the use of immobilized glycolipids in analytical high-performance liquid affinity chromatographic methods may serve as models in the study of interactions between gangliosides and carbohydrate-binding proteins. PMID- 8408436 TI - Characterisation of the photochemotherapeutic agent disulphonated aluminium phthalocyanine and its high-performance liquid chromatographic separated components. AB - Disulphonated aluminium phthalocyanine (AlPcS2), a potential clinical photosensitiser, has been synthesised in a reproducible form and shown by reversed-phase HPLC to consist of at least eight components which are believed to be individual AlPcS2 regioisomers. These components have been isolated either as single bands or mixtures of two using preparative reversed-phase HPLC methods. The number and position of sulphonate groups per phthalocyanine macromolecule for each component has been determined using a chemical degradation and HPLC assay. Results suggest that the bulk AlPcS2 material consists mostly (> 60%) of an amphiphilic alpha-, alpha-disubstituted regioisomer, with both sulphonate groups substituted to the same side of the molecule (adjacent form). Possible structures for some of the other separated components of AlPcS2 are also presented. PMID- 8408437 TI - The analysis of multiple phosphoseryl-containing casein peptides using capillary zone electrophoresis. AB - Multiple phosphoseryl-containing sequences of peptides and proteins stabilize amorphous calcium phosphate at neutral and alkaline pH and have been implicated in the nucleation/regulation of biomineralization. In an approach to analyze these peptides using capillary zone electrophoresis (CZE) we have attempted to relate the absolute electrophoretic mobility of various casein phosphopeptides to their physicochemical properties. Multiple phosphoseryl-containing peptides were selectively precipitated from enzymic digests of sodium caseinate and further purified using RP-HPLC and anion-exchange fast protein liquid chromatography. Purified fractions were then analyzed by CZE. Absolute electrophoretic mobilities of 13 peptides were determined by measurement of migration times relative to that of a neutral marker, mesityl oxide. A linear relationship (r2 = 0.993) was obtained between absolute electrophoretic mobility and q/M(r)2/3 where q is the net negative charge of the peptide calculated using relevant pKa values and M(r) is the molecular mass. M(r)2/3 is a measure of the surface area of a sphere that has a volume proportional to the M(r) of the peptide and relates to the frictional drag exerted on the peptide during electrophoretic migration. As absolute electrophoretic mobility is influenced by charge and size CZE can be used to monitor peptide phosphorylation, dephosphorylation, deamidation and truncation. This technique therefore would be suitable for quantitative analysis of peptide substrates in kinase and phosphatase studies. In conclusion CZE is a rapid and efficient technique for the resolution of multiple phosphoseryl containing peptides from enzymic digests of casein. PMID- 8408438 TI - Determination of penicillin G, ampicillin, amoxicillin, cloxacillin and cephapirin by high-performance liquid chromatography-electrospray mass spectrometry. AB - This report contributes to a preliminary investigation of high-performance liquid chromatographic (HPLC)-mass spectrometric (MS) methods for confirming beta-lactam antibiotic residues in bovine milk. Initial work for each antibiotic evaluated the collisional activated dissociation (CAD) spectra that could be generated between the capillary and skimmer in the electrospray (ESP) interface. The drugs show various characteristic fragmentation, mostly within the beta-lactam ring and the amide group. Response for a particular compound in a given solvent can vary drastically. Usually, the more organic component in the solvent, the higher the ESP response. In many cases use of acetonitrile also results in slightly better ion currents than for methanol when comparing equal percentages of either organic solvent in water. The ESP response of most of the tested antibiotics can be enhanced by the addition of formic acid or acetic acid to the mobile phase methanol-water (1:1). In general, the negative ion spectra are lower in intensity, exhibiting an [M-H]- ion and producing less fragmentation at higher CAD voltages as compared to positive ion spectra. An isocratic reversed-phase HPLC method for the separation of a mixture of five common beta-lactam antibiotics was developed using acetic acid as a mobile phase additive and optimized for detection with a new ESP HPLC-MS interface. A post-column split ratio of 70:1 for the eluent from a 150 x 2 mm I.D. column was chosen to provide the required lower flow-rate (approximately 4 microliters/min). The limit of detection for the simultaneous determination of these antibiotics was estimated to be 100 ppb. Electrospray HPLC-MS could be used to confirm these antibiotics for quantities down to about 100 pg entering the mass spectrometer. Multiresidue analysis with microbore HPLC-ESP-MS has the advantage that no post-column splitting of the eluent is required and all of the analyte (on-column injected) will be transferred into the ESP interface. Preliminary work showed good mass spectrometric sensitivity down to the level of regulatory interest, but chromatographic separation efficiency must be improved. PMID- 8408439 TI - Peptide mapping of recombinant human interferon-gamma by reversed-phase liquid chromatography with on-line identification by thermospray mass spectrometry and UV absorption spectrometry. AB - The detection and identification of minor peaks in a complex peptide map of recombinant human interferon-gamma was realized by on-line analysis of the eluted peptides using thermospray mass spectrometry and UV absorbance spectrometry. By this procedure the time-consuming process of collection, purification and chemical sequence analysis is avoided. Owing to the formation of multiple charged ions, the domain of the covered masses is extended. Fragmentation of the peptides in the thermospray source was observed resulting from, amongst others, cleavage by acid hydrolysis of peptide bonds involving an aspartic acid. This was of great use for the identification of peptides in a digest of recombinant human interferon-gamma by Staphylococcus aureus strain V8 endoprotease. PMID- 8408440 TI - Ion chromatographic analysis of glucose, fructose, and sucrose concentrations in raw and processed vegetables. AB - To meet the demands for improved carbohydrate analysis techniques, a method of sugar extraction and analysis has been developed using ion chromatography. The sample preparation method, which uses filtration and extraction steps, has been shown to adequately isolate glucose, fructose, and sucrose from other food components in raw and processed vegetables. Resulting sample preparations were successfully analyzed by ion chromatography using pulsed amperometric detection, which is a sensitive, rapid, and accurate technique. Sugar concentrations in a number of different vegetable products were successfully determined in this way. PMID- 8408441 TI - Ion chromatography on a new moderate capacity anion exchange column. AB - The mechanisms of retention of anions on a new solvent-compatible, moderate capacity (170 microequivalent per column) anion-exchange column are described. Retention based on anion exchange is shown using both sodium hydroxide and 4 cyanophenolate eluants. Plots of the log k' vs. log C, (where k' is the capacity factor and C is the eluant concentration) are shown to be linear with slopes approximating the charge of the eluant as predicted by the standard anion exchange equation. Sodium hydroxide is shown to be a relatively weak eluant, capable of separating monovalent anions that are difficult to separate on other columns. Sodium 4-cyanophenolate is shown to be a strong eluant, capable of eluting polyvalent anions. The retention of nonionic alkylphenones is also described. In plots of log k' vs. percentage acetonitrile in the eluant, these compounds were found not to follow the solvophobic mechanism typical of reverse phase HPLC columns. Results are compared with other anion exchange columns. Applications include the determination of lysophosphatidyl inositol in brain microsomes and methacrylic acid in esterase-treated samples. PMID- 8408442 TI - An avian hepatoma cell line for the cultivation of infectious laryngotracheitis virus and for the expression of foreign genes with a mammalian promoter. AB - Infectious laryngotracheitis virus (ILTV) is the causative agent of a highly infectious upper respiratory tract disease in chickens. Vaccine development and basic studies on ILTV have been hampered by the lack of a cell line for the cultivation of this herpesvirus which was identified in 1930. Four different avian cell lines were tested for their suitability to propagate ILTV. Here we report the successful growth of ILTV with a chemically-induced avian hepatoma cell line, while retrovirus transformed cell lines derived from permissive primary cells, were found to be non-permissive for ILTV. After multiple passaging of ILTV in the hepatoma cells, the virus could be grown up to a titre of 1 x 10(7) EID50 per ml with a replication cycle comparable to that in primary hepatocytes. Methods of plaque assay, DNA-transfection, and expression of a reporter gene were established. The gene coding for the bacterial beta galactosidase gene under the control of the cytomegalovirus (CMV) immediate-early promotor was transiently expressed, indicating that a mammalian herpesvirus promotor was recognized by this avian cell line. Infectious ILTV virions were produced after transfecting this cell-line with purified ILTV DNA. The results indicated that the cell line is suitable for the construction of recombinant ILTV and for the molecular biological study of this important avian pathogen. PMID- 8408443 TI - Time course of hepatitis A antibody production after active, passive and active/passive immunisation: the results are highly dependent on the antibody test system used. AB - Two commercially available automated test systems for hepatitis A antibody, HAVAB IMX (Abbott) and ENZYMUN Anti-HAV (Boehringer) were evaluated in a study of active, passive and active/passive immunisation against hepatitis A. The inactivated hepatitis A vaccine Epaxal Berna and the hepatitis A immunoglobulin preparation Globuman were products of the Swiss Serum and Vaccine Institute. Although both hepatitis A antibody test kits were standardised with the same international WHO standard hepatitis A immunoglobulin preparation, divergent results were obtained for the level of circulating hepatitis A antibody after vaccination. One month after the vaccination the mean geometric antibody titres were 315 mIU/ml after active, 253 mIU after active/passive and 22 mIU after passive immunisation when measured with the Enzymun assay. In the same sera 70 mIU/ml after active, 60 mIU after active/passive and 18 mIU after passive immunisation could be detected with the IMX test. Antibody avidity studies could not explain the differences obtained by the two test methods. The neutralization test is the standard method for the estimation of protection against hepatitis A. This test is not suitable for large series of serum samples, and enzyme immunoassays are indispensable for vaccination studies. To be suitable for monitoring antibody development in phase I and II clinical trials as well as in postmarketing studies, EIA tests for hepatitis A antibodies must be commercially available and of known sensitivity. The Enzymun anti-HAV test developed by Boehringer Mannheim (Germany) offers the possibility to measure antibody titres around the protective level of 20 mIU/ml which is reached by the passive immunisation with immunoglobulin preparations or within two weeks after active vaccination with an inactivated hepatitis A vaccine. The Abbott IMX test system is more useful for the detection of natural infections by the hepatitis A virus. PMID- 8408444 TI - High-performance liquid chromatography to assess the effect of serum storage conditions on the detection of hepatitis C virus by the polymerase chain reaction. AB - The effect of various serum storage conditions on the detection of hepatitis C virus (HCV) by the polymerase chain reaction (PCR) was assessed. 50 microliters aliquots of serum from four HCV PCR positive patients were subjected, in triplicate, to: (a) storage at -20 degrees C for 1, 5, 10 days; (b) storage at room temperature (RT) for 1, 2, 7 days; (c) 1, 3, 5, and 10 successive freeze thaw cycles; (d) incubation at 37 degrees C, and 56 degrees C for 1, 3, 24 h; and (e) storage in guanidium-thiocyanate extraction buffer at RT, and 4 degrees C for 1, 5, 10 days. PCR products were detected by agarose gel electrophoresis (AGE) and quantitatively by high-performance liquid chromatography (HPLC). Only storage in extraction buffer at RT for 5-10 days and incubation at 56 degrees C for 24 h appeared to result in a loss of > or = 50% of detectable HCV PCR product. Up to 10 successive freeze-thaw cycles, storage at -20 degrees C for up to 10 days or at RT for up to 7 days, storage in extraction buffer at RT for 1 day or at 4 degrees C for up to 10 days, and incubation at 37 degrees C for up to 24 h resulted in minimal PCR signal loss. HPLC was a reproducible method of detecting and quantitating HCV PCR products, and was more sensitive than AGE. PMID- 8408445 TI - Detection of bovine viral diarrhea virus RNA in formalin-fixed, paraffin-embedded brain tissue by nested polymerase chain reaction. AB - Isolation and amplification of RNA from formalin-fixed, paraffin-embedded tissues is delicate due to its fragility and ubiquitous ribonucleases. For retrospective studies, however, a convenient procedure for the detection of RNA in archived material is of great value. Bovine viral diarrhea (BVD) virus is a member of the pestivirus genus in the family Flaviviridae. Different protocols for the isolation of BVD virus RNA from fresh and autolytic as well as from routinely formalin-fixed and paraffin-embedded brain tissue of BVDV-infected calves were compared. The polymerase chain reaction (PCR) after reverse transcription (RT PCR) was carried out subsequently for the detection of viral RNA. Using proteinase K digestion of deparaffinized tissue sections without additional ribonuclease inhibitors and subsequent nested PCR, a 803 bp fragment of the gene coding for the nonstructural protein p125 of BVD virus could be consistently detected. In addition, BVD virus RNA was detected by RT-PCR from non-fixed brain tissue after 10 days of autolysis. PMID- 8408446 TI - Using the polymerase chain reaction to genotype human papillomavirus DNAs in samples containing multiple HPVs may produce inaccurate results. AB - Compared with other laboratory techniques, the polymerase chain reaction (PCR) is a simple, rapid, sensitive method for detecting human papillomavirus (HPV) DNA in cervical samples. However, since many cervical samples contain multiple HPV types, we decided to investigate whether PCR results from such samples accurately reflected the relative amounts of each HPV type present. Theoretical calculations of product accumulation when multiple DNAs with different amplification efficiencies are present in the same sample were done. In addition a set of samples in which cloned HPV DNAs were mixed in varying proportions prior to PCR was tested. Finally, four clinical samples containing multiple HPV types by hybridization assays were subjected to PCR, using two different primer sets. Each of these lines of investigation showed that selective amplification of one HPV DNA over another can occur when mixed HPV types are present. This effect may lead to inaccurate information regarding both types and relative amounts of HPV DNAs in samples containing multiple HPV types. A protocol to avoid this problem is described. PMID- 8408447 TI - Direct sequencing of consensus primer generated PCR fragments of human papillomaviruses. AB - Consensus primers specific for the L1 and E1 regions were used to amplify HPV DNA fragments. These fragments were then directly sequenced using the consensus primers as the sequencing primers. The linking of PCR amplification, direct sequencing, and computer data bank analysis provides a new approach in the detection of different HPVs and has several advantages over existing methodology. These advantages include increased precision in the rapid characterization of known HPVs, detection of mutations, and identification of new HPV types. PMID- 8408448 TI - Steroid-modulated stromal cell tissue factor expression: a model for the regulation of endometrial hemostasis and menstruation. AB - This study examined steroidal regulation of tissue factor expression by cultured endometrial stromal cells. Confluent stromal cell cultures derived from cycling human endometria were exposed to vehicle control, 10(-8) mol/L estradiol (E2), 10(-8)-10(-6) mol/L medroxyprogesterone acetate (MPA), or both E2 and MPA for 2 24 days in serum-containing medium. The progestin enhanced immunoreactive and functionally active stromal cell tissue factor content, achieving peak effects by 8-12 days of culture. Although E2 alone was ineffective, it augmented MPA enhanced tissue factor content by 8 days of culture, with continued increases beyond 20 days. Dose-dependent effects on tissue factor protein content were observed between 10(-8)-10(-6) mol/L MPA added alone or together with E2. The content of tissue factor mRNA was also increased by MPA and synergistically increased by E2 plus MPA. Similar steroidal effects on stromal cell tissue factor protein and mRNA content were observed using a defined medium. After optimal induction of tissue factor expression by E2 plus MPA, removal of these steroids reduced levels of stromal cell tissue factor mRNA and protein, with virtually complete reversal by day 7 of withdrawal. These time-course and dose-response relationships establish in vitro conditions with which to dissect factors controlling endometrial hemostasis, whereas the observed effects of steroid withdrawal establish a novel model for the study of mechanisms regulating normal and abnormal uterine bleeding. PMID- 8408449 TI - Reduced response to metoclopramide and anomalous rising response to upright posture of plasma aldosterone concentration in Japanese patients with aldosterone producing adenoma. AB - We examined the changes in plasma aldosterone concentration (PAC) during 3 kinds of postural stimulation tests and a metoclopramide test in 20 Japanese patients with surgically proven aldosterone-producing adenoma. In the 2-h upright test (n = 16), PAC increased (P < 0.01) in 14 of 16 patients, with a mean percent change of 135 +/- 42% (+/- SD) of the supine levels without any significant change in plasma cortisol. In the furosemide-upright test (n = 17), PAC increased (P < 0.01) in 15 of 17 patients by 131 +/- 27%, with a significant increase in cortisol. However, in the furosemide upright test after dexamethasone pretreatment, which suppressed plasma cortisol during the test, PAC also increased (P < 0.01) in all 9 patients examined by 158 +/- 36%. Angiotensin-II infusion failed to increase PAC in all 4 patients examined whose PAC increased with postural stimulation tests. Injection of 10 mg metoclopramide (n = 13) induced a significant, but less marked, increase (P < 0.01) in PAC to 157 +/- 59% of basal levels, which was lower than those in previously reported series in Western countries. These results suggest that 1) there is a difference in PAC responsiveness to postural stimulation tests and to metoclopramide injection between Japanese and Western patients; and 2) the difference in the metoclopramide-induced aldosterone response implies a relative reduction of tonic dopaminergic inhibition of aldosterone secretion in our Japanese patients with aldosterone-producing adenoma. PMID- 8408450 TI - Body composition and muscle strength in healthy men receiving testosterone enanthate for contraception. AB - To determine the effect of androgens on body composition and muscle strength, we measured fat-free mass (kg), fat mass (kg), and bone density (g/cm2) by dual x ray absorptiometry, and muscle strength (Newton meters) by dynamometry in a controlled, prospective study involving 13 nonathletic men receiving testosterone enanthate 200 mg/week in for 6 months and 8 healthy controls. Biochemical markers of bone turnover were measured in the treated subjects at baseline and 6 months. In the treated subjects at 6 months, fat-free mass (mean +/- SEM) increased by 9.6 +/- 1.0% (P < or = 0.01) whereas fat mass decreased by 16.2 +/- 6.7% (P < or = 0.05). Changes in muscle strength ranged from -1.6-19.2%. Only hip adduction increased 19.2 +/- 9.5% (P < 0.05). Changes in bone density ranged from -1.3 5.2%, decreasing significantly at one site and increasing significantly at four of the nine sites measured (P < 0.05). Serum testosterone increased by 91.1 +/- 7.5% (P < 0.01), and testicular volume decreased by 24.0 +/- 3.2% (P < 0.01). Serum osteocalcin increased by 35.7 +/- 17.3% (P < 0.05), serum immunoreactive PTH (iPTH) increased by 41.4 +/- 15.1% (P < 0.05), serum calcium decreased by 2.3 +/- 1.0% (P < 0.05), and serum albumin decreased by 4.5 +/- 1.7% (P < 0.05). There were no detectable changes in fat-free mass, fat mass, muscle strength, or bone density in controls. The administration of testosterone enanthate in pharmacological doses for 6 months resulted in a modest reduction in fat mass and small increases in fat-free mass, muscle strength, and bone density. These changes do not support the use of androgens for enhancing athletic performance. PMID- 8408451 TI - Evidence for two differentially regulated populations of peripheral beta endorphin-releasing cells in humans. AB - In the current study we tested the hypothesis that human plasma beta-endorphin (beta E) is derived from at least two subpopulations of beta E-releasing cells: one sensitive to glucocorticoids as well as to dopamine (DA; regulated analogously to the corticotrophs of the rat pituitary), and one insensitive to glucocorticoids but sensitive to DA (regulated analogously to the melanotrophs of the rat pituitary). To test this hypothesis, human plasma levels of ACTH, cortisol, and beta E-like immunoreactivity were measured at baseline and after haloperidol treatment (0.05 mg/kg, i.v.) in two experimental groups, one pretreated with dexamethasone (1.5 mg) and one pretreated with placebo. Plasma PRL levels were also measured in both groups as an indicator of DA receptor blockade. Dexamethasone partially suppressed both baseline and haloperidol stimulated levels of human plasma beta E-like immunoreactivity, whereas it completely suppressed both basal and haloperidol-stimulated levels of ACTH and cortisol and had no statistically significant effect on either basal or haloperidol-stimulated PRL levels. These data support a negative feedback effect of glucocorticoids on one DA-sensitive cell population that releases both ACTH and beta E (corticotroph like), but not on a second cell population that releases beta E but not ACTH. PMID- 8408452 TI - Determination of estrogen receptor messenger ribonucleic acid (mRNA) and cytochrome P450 aromatase mRNA levels in adipocytes and adipose stromal cells by competitive polymerase chain reaction amplification. AB - Clinical evidence suggests that sex hormones affect adipose tissue metabolism and deposition. To investigate the biosynthesis and possible action of estrogen in adipose tissue, we report the use of competitive, specific polymerase chain reaction amplifications to determine levels of estrogen receptor (ER) messenger RNA (mRNA) and cytochrome P450 aromatase mRNA in adipocytes and adipose stromal cells. This extremely sensitive technique uses coamplification of a homologous animal species complementary RNA to control for differences in amplification efficiencies. The DNA amplification products are identified by Southern hybridization with species-specific radiolabeled oligonucleotide probes. Abdominal adipose tissue obtained from female patients during elective abdominoplasty was separated by collagenase digestion and centrifugation into floating adipocytes and pelleted adipose stromal cells. Our results demonstrate higher ER mRNA levels in adipocytes compared to adipose stromal cells, whereas cytochrome P450 aromatase mRNA levels are higher in adipose stromal cells compared to adipocytes. The finding of ER mRNA in adipose tissue suggests the presence of the ER in adipose tissue. In addition the inverse correlation of ER mRNA and cytochrome P450 aromatase mRNA levels in adipocytes and adipose stromal cells suggests a paracrine relationship whereby estrogen produced by adipose stromal cells affects adjacent adipocytes. PMID- 8408453 TI - p53 mutations in all stages of thyroid carcinomas. AB - The p53 gene has been implicated as a tumor suppressor gene whose inactivation by mutations has been noted in a variety of human malignancies. Using single strand conformation polymorphism analysis of cDNA fragments amplified by reverse transcription-polymerase chain reaction, we analyzed 57 thyroid tumor specimens (8 follicular adenomas and 49 carcinomas) for the presence of mutations in exons 5, 6, 7, and 8 of p53 gene. Twelve of 49 (24.5%) of the thyroid carcinomas tested presented a mutated p53 allele, but none of the 8 benign thyroid tumors did. Mutations were found in 1 of 5 anaplastic carcinomas and 11 of 44 differentiated carcinomas. Three of these 11 differentiated tumor specimens showed foci of solid tissue with evidence of dedifferentiation. Two samples (1 with anaplastic carcinoma, the other with papillary carcinoma) had double mutations on the same allele resulting in a frameshift. Most mutations were point mutations, and 50% of those were G:C to A:T transitions. Seventy-five percent of the mutations were in exons 7 and 8. The presence of p53 mutations was not associated with tumor stage or histological type. Our data suggest that p53 mutations are involved in thyroid carcinogenesis and may play an important role in the malignant transformation of thyroid cells as well as thyroid tumor progression. PMID- 8408454 TI - Acute changes in calcium homeostasis during treatment of primary hyperparathyroidism with risedronate. AB - We administered risedronate, a potent oral bisphosphonate, to patients with mild primary hyperparathyroidism in order to 1) determine if we could normalize the serum calcium concentration in the short term, and 2) analyze changes in the homeostatic mechanisms responsible for maintaining hypercalcemia in this patient population. When administered for 7 days, risedronate reduced fasting serum calcium concentrations without significant toxicity in patients with primary hyperparathyroidism. The decrease in serum calcium was accompanied by evidence of inhibition of bone resorption, as assessed by measurement of urinary hydroxyproline, increased serum immunoreactive PTH concentrations, enhanced renal tubular reabsorption of calcium, and a progressive decrease in serum alkaline phosphatase. Serum PTH was partially suppressed by an oral calcium load in untreated patients as well as in patients treated with risedronate. Although patients treated with risedronate had normal fasting serum calcium levels, serum calcium values in these normocalcemic patients were labile after oral ingestion of calcium. After daily calcium intake of 2 g, serum calcium levels in risedronate-treated patients were similar to those in untreated patients with primary hyperparathyroidism, suggesting that there are likely to be fluctuations in serum calcium in risedronate-treated patients with normal fasting serum calcium during postprandial periods. These studies show that risedronate lowers fasting serum calcium during short term treatment. However, further studies are required to determine whether the lability in serum calcium in these patients after an oral calcium load has clinical significance, and whether longer term treatment would maintain serum calcium in the normal range. PMID- 8408455 TI - In vitro production of interleukin-1, interleukin-6, and tumor necrosis factor alpha in insulin-dependent diabetes mellitus. AB - The in vitro production of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) by monocytes was examined in patients with insulin-dependent diabetes mellitus (IDDM), in those with noninsulin-dependent diabetes mellitus (NIDDM), and in healthy volunteers. The production of IL-1 and IL-6 by monocytes was significantly lower in IDDM patients than in NIDDM patients and normal subjects whereas the TNF-alpha production by monocytes did not differ between IDDM patients and normal subjects. On the other hand, the TNF-alpha production was significantly higher in NIDDM patients than in IDDM patients and normal subjects. There was a significant correlation between IL-1 and IL-6 concentrations in culture supernatants of monocytes for IDDM patients but not for NIDDM patients and normal subjects. Neither glucose nor insulin showed any stimulatory effect on in vitro production of these monokines. In the serial observation lasting 3-18 months, the monocyte production of IL-1 was found to be consistently reduced in IDDM patients unrelated to the control state of diabetes, suggesting that the reduction of the IL-1 and IL-6 production by monocytes in IDDM patients may be intrinsically affected by immunological defects. PMID- 8408456 TI - Amelioration of some pregnancy-associated variations in thyroid function by iodine supplementation. AB - Knowledge of the effect of differences in iodine intake levels on public health in areas with no endemic goiter is limited. Groups at risk when iodine intake is relatively low are pregnant and lactating women and their newborns. A prospective randomized study was performed to evaluate the effect of iodine supplementation in an area where the median daily iodine excretion in urine is around 50 micrograms. Fifty-four normal pregnant women were randomized to be controls or to receive 200 micrograms iodine/day from weeks 17-18 of pregnancy until 12 months after delivery. In the control group, serum TSH, serum thyroglobulin (Tg), and thyroid size showed significant increases during pregnancy. These variations were ameliorated by iodine supplementation. Iodine did not induce significant variations in serum T4, T3, or free T4. Cord blood Tg was much lower when the mother had received iodine, whereas TSH, T4, T3, and free T4 levels were unaltered. The results suggest that a relatively low iodine intake during pregnancy leads to thyroidal stress, with increases in Tg release and thyroid size. However, the thyroid gland is able to adapt and keep thyroid hormones in the mother and the child normal, at least under normal circumstances, as evaluated in the present study. It is not known whether this stress is sufficient to be of importance for late development of autonomous thyroid growth and function. PMID- 8408457 TI - Interferon-gamma inhibition of human thyrotropin receptor gene expression. AB - To evaluate the role of interferon-gamma (IFN gamma) on human thyroid-specific gene expression, the effect of IFN gamma on TSH- and cAMP-induced TSH receptor gene expression was studied using cultured thyroid cells obtained from normal thyroid glands and those from patients with Graves' disease. Incubation of Graves' thyroid cells with 1.0 U/L bovine TSH or 1.0 mM 8-bromo-cAMP resulted in a 2-fold increase in TSH receptor mRNA expression, which was markedly inhibited in the presence of IFN gamma in a dose- and time-dependent manner. This inhibitory effect was completely neutralized by monoclonal antibody against IFN gamma. IFN alpha and -beta had no influence on TSH- and cAMP-stimulated TSH receptor mRNA expression. Paranodular normal thyroid cells showed the same results as those obtained using Graves' thyroid cells. Scatchard analysis of the [125I]TSH binding study showed that IFN gamma inhibited the number of TSH receptors up-regulated by TSH on the cell surface at the low affinity binding site (4.1 vs. 8.2 x 10(5)/cell). These results indicate that IFN gamma suppresses TSH- and cAMP stimulated human TSH receptor gene expression, resulting in a decrease in the number of TSH receptors. In conclusion, IFN gamma interacts via an intermediate pathway of TSH signal transduction and attenuates TSH receptor synthesis in normal and Graves' thyroid cells. PMID- 8408458 TI - Uptake of 18-fluoro-2-deoxy-D-glucose by thyroid cancer: implications for diagnosis and therapy. AB - A patient developed a pulmonary metastasis from papillary thyroid carcinoma. This tumor concentrated relatively little 131I, but sufficient 18F-fluoro-2-deoxy-D glucose (FDG) to be quantified and imaged by positron emission tomography. The uptake of FDG was lower on positron emission tomographic images after T4 therapy and when the serum TSH concentration was reduced to the low normal range. It may be possible to use decreases in FDG uptake by thyroid cancers, which represent declines in metabolism by the tumors, to indicate the optimum doses of T4 treatment for patients with these neoplasms. In addition, the ratio of tumor to background radioactivity was higher for FDG than for the flow agent 201Tl, so that studies with FDG may be a useful scintigraphic method for locating thyroid cancers when radioiodine imaging is unsatisfactory. PMID- 8408459 TI - Antioxidant potential of specific estrogens on lipid peroxidation. AB - Coronary heart disease (CHD) is the leading cause of death in postmenopause. Estrogen administration in postmenopause lowers the risk of CHD by 50%. A variety of estrogen preparations are currently used in postmenopausal hormone replacement therapy. It is unknown, however, if structural differences in the estrogen molecule influence the cardioprotective effects of estrogens. In this communication we have shown that equine estrogens (especially equilin) exhibit higher antioxidant potency (as measured by fatty acids and sterols oxidation) when compared to estrone and estradiol-17 beta. PMID- 8408460 TI - Differential autoantibody responses to thyroid peroxidase in patients with Graves' disease and Hashimoto's thyroiditis. AB - In order to examine the specificity of the autoantibody response to thyroid peroxidase (TPO, previously identified as thyroid microsomal antigen) in autoimmune thyroid disease, we examined reactivity of sera from 45 Hashimoto's and 48 Graves' patients to native thyroid microsomes, denatured and reduced human TPO and several recombinant fragments of human TPO corresponding to amino acids 457-933 of the native protein. Both Graves' and Hashimoto's sera bound native, denatured and reduced TPO at significantly greater rate than normal controls, and no differences were noted between the two disorders in binding to these forms of the autoantigen. Binding was also noted to two recombinant fragments of TPO, corresponding to amino acids 513-633 and 633-933 in TPO. The frequency of autoantibodies to the TPO AA(633-933) region was not significantly different in Hashimoto's vs. Graves' disease patients (58% vs. 65% respectively), and appeared to relate to evidence of glandular inflammation in the Graves' patients (presence of anti-thyroglobulin antibodies and elevated anti-microsomal antibody levels). In contrast, antibodies to the TPO AA(513-633) fragment were significantly more common and of higher titer in Hashimoto's vs. Graves' disease patients, and did not correlate with any measure of glandular inflammation. These results identify two specific regions of TPO autoantibody binding and indicate that there are differences in the autoantibody response to TPO in Hashimoto's and Graves' diseases. PMID- 8408461 TI - Clinical review 50: Clinically silent adrenal masses. PMID- 8408462 TI - Thyrotropin-releasing hormone stimulation tests in infants. AB - The TSH response to TRH administration (7 micrograms/kg) was measured in 68 infants (22 premature) who had abnormal thyroid screening tests by the filter paper method and whose serum thyroid function tests were only mildly abnormal. Twenty-eight infants (12 premature) had peak TSH values of 35 mU/L or less and were considered normal (group I). Forty infants (10 premature) had peak TSH values above 35 mU/L and were considered hyperresponsive (group II). The mean age at testing, screening T4, TSH levels that prompted the testing, as well as baseline T4, T3, and free T4 at the time of TRH testing were not different between the groups. The mean (+/- SD) baseline TSH value was greater in group II (6.8 +/- 2.3 mU/L) than in group I (4.4 +/- 2.2 mU/L; P < 0.001). However, there was a great deal of overlap in the individual TSH values (group I, 0.9-10 mU/L; group II, 1.9-10.6 mU/L). Mean peak TSH levels were significantly different in the two groups (group I, 24 +/- 7.7 mU/L; group II, 60.3 +/- 26.1 mU/L; P < 0.001). During long term follow-up, all 25 group I infants available for evaluation have been confirmed as clinically and biochemically normal. No infant diagnosed as normal was later found to have evidence of hypothyroidism. Fourteen infants in group II have had evidence of thyroid dysfunction. We conclude that the TSH response to TRH stimulation is a useful tool for the evaluation of infants suspected of having primary hypothyroidism. Whether hyperresponsiveness to TRH represents a form of neonatal hypothyroidism requiring treatment remains to be determined. PMID- 8408463 TI - Effects of acute mineralocorticoid and glucocorticoid receptor blockade on the excretion of an acute potassium load in healthy humans. AB - To examine the role of mineralocorticoid and glucocorticoid in potassium (K) tolerance in healthy humans, we studied the effects of canrenoate, a mineralocorticoid antagonist, and RU486, a glucocorticoid antagonist, on the excretion of a KCl load. Canrenoate (200 mg, iv) or RU486 (400 mg, orally) was administered 150 min before a KCl load (1 mmol/kg BW, orally) in seven healthy males undergoing maximal water diuresis. Clearance studies were extended for 5 h after the KCl load, and the data were compared with time control, KCl load alone, and canrenoate alone. KCl increased K excretion (from 18.8 +/- 2.4 to 63.3 +/- 3.9 mmol/5 h; P < 0.01) and sodium (Na) excretion (from 35.9 +/- 2.1 to 72.9 +/- 6.0 mmol/5 h; P < 0.01). Clearance calculations, based on maximal water diuresis, were compatible with increased distal Na and volume delivery. Canrenoate alone modestly increased basal cumulative NaCl excretion and had no effect on K excretion. However, canrenoate blunted the kaliuresis after the KCl load (51.9 +/ 4.4 mmol/5 h; P < 0.05 compared to KCl alone) and stimulated natriuresis in a complementary way. Clearance data were compatible with diminished distal Na reabsorption and K secretion in response to an undisturbed KCl-induced increase in distal Na delivery. RU486 did not influence the excretion of the KCl load or its effects on renal sodium handling parameters, although effective glucocorticoid receptor blockade was likely to be present in view of the increase in plasma cortisol. These data suggest that in healthy humans, mineralocorticoid activity, but not glucocorticoid activity, is involved in the elimination of a K load. The latter contrasts with data in adrenalectomized animals, in which situation glucocorticoid as well as aldosterone are indispensible for normal K tolerance. PMID- 8408464 TI - The effect of the hypoestrogenic state, induced by gonadotropin-releasing hormone agonist, on Doppler-derived parameters of aortic flow. AB - The effect of the hypoestrogenic state, induced by a GnRH agonist (GnRH-a), on cardiac function in healthy young women, was evaluated by Doppler echocardiography performed before treatment and when serum 17 beta-estradiol levels were suppressed by GnRH-a to 36.7 pmol/L. The following parameters of aortic flow were measured: peak flow velocity, ejection time, and acceleration time. Additional parameters calculated were flow velocity integral, cardiac index, and mean acceleration. The study group included 15 menstruating women, aged 25-42 yr (mean, 33 yr), with symptomatic fibroids, endometriosis, or scheduled for in vitro fertilization, who were treated with a GnRH-a. There were significant decreases in peak flow velocity (99 +/- 11 vs. 86 +/- 11 cm/s; P = 0.0004) and cardiac index (3.0 +/- 0.7 vs. 2.5 +/- 0.5 L/min.m2; P = 0.002). A decrease that did not reach statistical significance was noted in flow velocity integral (18.9 +/- 2.7 vs. 16.5 +/- 3.4 cm; P = 0.07). Mean acceleration was decreased significantly (12.6 +/- 2.6 vs. 10.8 +/- 1.8 m/s.s; P = 0.01), but no significant changes in acceleration time (81 +/- 16 vs. 83 +/- 10 ms; P = 0.7) or ejection time (296 +/- 25 vs. 295 +/- 27 ms; P = 0.8) were observed. These results indicate that estrogen deprivation is associated with smaller stroke volume and flow acceleration and might suggest that hypoestrogenism has a direct effect on cardiovascular performance. PMID- 8408465 TI - The effects of an antiprogestin, mifepristone, and an antiestrogen, tamoxifen, on endometrial 17 beta-hydroxysteroid dehydrogenase and progestin and estrogen receptors during the luteal phase of the menstrual cycle: an immunohistochemical study. AB - The effects of a progesterone antagonist, mifepristone (RU486), and an estrogen antagonist, tamoxifen, given during the early luteal phase on endometrial 17 beta hydroxysteroid dehydrogenase (17HSD) and estrogen (ER) and progesterone (PR) receptors were studied. Eleven regularly menstruating women were studied during control and treatment cycles. In the treatment cycle on day LH + 2 (2 days after the peak serum LH concentration), 10 subjects received a single dose of 200 mg mifepristone, and 9 received 2 doses of 40 mg tamoxifen on days LH + 2 and LH + 3. In addition, 4 subjects received 400 mg mifepristone in a separate treatment cycle. 17HSD, ER, and PR were measured immunohistochemically in endometrial tissue specimens taken on days LH + 6 to LH + 8. Blood samples were conducted during control and treatment cycles, and serum estradiol, progesterone, and LH concentrations were quantified by RIA. Administration of mifepristone blocked the induction of 17HSD by progesterone and prevented the expression of 17HSD in gland and surface epithelial cells in 8 patients. In 2 patients, staining of 17HSD was seen during both the control and mifepristone treatment cycles. The higher dose of mifepristone additionally given to four subjects did not block the expression of 17HSD in 2 cases where blocking was observed with the lower dose of mifepristone, and in 1 of these patients, very strong staining of 17HSD was observed in basal cells beneath the epithelial cells. ER and PR showed intense staining in the nuclei of both gland and stromal cells in mifepristone treatment cycles, whereas receptor staining was faint or absent in the respective control cycles. Tamoxifen did not have any significant effect on staining of 17HSD or the abundance of receptors. Serum concentrations of estradiol, progesterone, and LH were not significantly affected by the administration of mifepristone or tamoxifen. This study reveals that mifepristone, administered in the early luteal phase, usually blocks the expression of 17HSD and the down-regulation of PR and ER. However, the expression of 17HSD in some patients may reflect the ineffectiveness of the mifepristone treatment used to prevent implantation in certain subjects. PMID- 8408466 TI - Corticotropin-releasing factor-binding protein is produced by human placenta and intrauterine tissues. AB - CRF circulates in high concentration in pregnant woman. It is produced by the placenta and the other intrauterine tissues (maternal decidua, amnion, and chorion). Recently, a CRF-binding protein (CRF-BP) has been identified and cloned. It binds the circulating CRF, reducing its biological action during pregnancy. Liver is the major source of CRF-BP. The aim of the present study was to evaluate whether human placenta and intrauterine tissues produce CRF-BP. The localization of mRNA and immune CRF-BP by in situ hybridization and immunohistochemistry, respectively, was performed. Antisense and sense riboprobes synthesized from a fragment of human CRF-BP cRNA and a specific rabbit anti-hCRF BP serum was used. The syncytial layer of placental villi at term intensely expressed CRF-BP mRNA and immunoreactivity, whereas rare positively hybridized cells were observed within the cytotrophoblasts and mesenchymal cells. Large decidual cells, amniotic epithelial cells, and chorionic cytotrophoblast stained positively for CRF-BP mRNA and protein. Control sections collected from the same tissues failed to show any positive localization of sense strand cRNA probe and antiserum preadsorbed with immunogen. Finally, the addition of recombinant CRF-BP to human cultured placental cells significantly decreased CRF-induced ACTH release, with a dose-dependent effect. The present data show that local production of CRF-BP occurs in human trophoblast and intrauterine tissues and may represent one of the major mechanisms used by targets tissues to control CRF activity during pregnancy. PMID- 8408467 TI - Regulation of inhibin secretion in human placental cell culture by epidermal growth factor, transforming growth factors, and activin. AB - In this study, the effects of epidermal growth factor (EGF), transforming growth factor-alpha (TGF alpha), TGF beta 1, and activin-A on inhibin secretion were investigated in primary culture of human placental cells. Immunoreactive inhibin in the culture medium was measured by immunoenzymatic assay. EGF stimulated testosterone-induced inhibin secretion in placental cells. Although testosterone alone induced only a slight enhancement of inhibin release in the culture, treatment of trophoblast cells with EGF and testosterone caused a significant increase in inhibin secretion, with immunoreactive inhibin levels much higher than those of testosterone or EGF alone. TGF alpha combined with human placental lactogen (hPL) had a stimulatory effect on inhibin secretion in placental cell culture. Simple addition of either TGF alpha or hPL to the culture did not show any effect on inhibin secretion in placental cells. A remarkable augmentation of inhibin secretion was obtained after the trophoblasts were exposed to both TGF alpha and hPL simultaneously. TGF beta 1 and activin-A showed synergistic effects to suppress inhibin secretion in placental cells. TGF beta 1 alone did not show any action on inhibin secretion, and activin-A alone induced a small decrease in inhibin release in the culture. In the presence of activin-A, addition of TGF beta 1 to the culture induced a profound decrease in immunoreactive inhibin levels in the medium. Activin-A could also suppress hCG-induced inhibin secretion in placental cells. Addition of hCG alone resulted in a small, but not significant, increase in inhibin release in the cultured cells, whereas the presence of activin-A combined with hCG in the culture conversely decreased inhibin secretion in the culture, with immunoreactive inhibin levels significantly lower than those in the presence of hCG or activin-A alone. These findings suggest that EGF and TGF alpha, alone or in combination with other hormones, may be stimulators, and TGF beta and activin may act as suppressors of inhibin secretion in human placental cells. PMID- 8408468 TI - Correlation of microsomal antibodies with the intensity of the intrathyroidal autoimmune process in Graves' disease. AB - Graves' disease is an organ-specific autoimmune disease, and intrathyroidal lymphocytes seem to be the major source of thyroid autoantibodies. Consequently, the intensity of the intrathyroidal lymphocytic infiltration is generally believed to reflect the activity of the autoimmune process. We, therefore, investigated the correlation of microsomal (enzyme immunoassay), thyroglobulin (RIA), and TSH receptor antibodies (RRA) with the degree of intrathyroidal infiltration by immunoglobulin G-producing plasma cells, activated T-cells, antigen-presenting cells, and the total number of lymphocytes. The immunocompetent cells were identified immunohistologically with monoclonal antibodies for immunoglobulins kappa and lambda, UCHL1, and the S100 antibody, respectively, in 26 thyroid glands of patients suffering from Graves' disease. The intensity of lymphocytic infiltration was determined by the point-counting method and by counting all lymphocytes and the labeled lymphocytes in 3 x 51 visual fields or 3 slides/thyroid gland. Microsomal antibodies correlated significantly (P = 0.001) with the total number of lymphocytes (r = 0.86), kappa (r = 0.71), lambda (r = 0.71), UCHL1 (r = 0.9), and S100 (r = 0.9) positive cells. These correlations were also significant for thyroglobulin antibodies. However, TSH receptor antibodies showed no significant correlations with any of the populations of immunocompetent cells. Patients with preoperatively undetectable TSH receptor or microsomal antibodies showed a broad variation of intrathyroidal infiltration by the immunocompetent cells investigated. Microsomal antibody titers, therefore, seem to reflect the intensity of the intrathyroidal autoimmune process in Graves' disease better than TSH receptor antibodies. However, the broad baseline variation in intrathyroidal infiltration observed with nondetectable thyroid antibodies will not always allow determination of the intensity of the intrathyroidal autoimmune process from microsomal or thyroglobulin antibody titers. PMID- 8408469 TI - The metabolic disposition of plasma 5 alpha-dihydroprogesterone (5 alpha-pregnane 3,20-dione) in women and men. AB - This study was conducted 1) to ascertain whether the high levels of plasma 5 alpha-dihydroprogesterone (5 alpha DHP) during the luteal phase of the human ovarian cycle and pregnancy are attributable to high rates of production or, alternatively, low rates of clearance, and 2) to estimate the relative distribution of the irreversible metabolism of 5 alpha DHP, i.e. hepatic compared with extrahepatic clearance of plasma 5 alpha DHP. The concentration of 5 alpha DHP in plasma of women during the luteal phase of the ovarian cycle and pregnancy is very high, viz. 12-40% that of progesterone. Thus, a potentially large source of steroid precursor is available for the formation of bioactive 5 alpha pregnanolone metabolites. We found that the MCR of 5 alpha DHP in women and men is 4187 +/- 312 L plasma/24 h (range, 3181-5506; n = 6). The MCR of 5 alpha DHP as a function of body surface area was 2406 +/- 240 L/24 h.m2. The MCR of 5 alpha DHP, therefore, is the greatest of any steroid reported, except for that of the catechol estrogens, which are metabolized intravascularly by erythrocyte catechol O-methyltransferase. Based on estimated rates of liver plasma flow (1500 L/24 h) and hepatic extraction (75-85%) of lipophilic steroids that are not specifically bound in plasma with high affinity to binding proteins as determined by other investigators (e.g. 5 alpha DHP), we estimate that approximately 1200 L plasma/24 h are cleared of 5 alpha DHP by liver and approximately 2800 L plasma/24 h are cleared of 5 alpha DHP (70%) by metabolism in extrahepatic tissues. Thus, 5 alpha DHP can serve as a precursor for bioactive 5 alpha-pregnanolone(s). PMID- 8408470 TI - Bone metabolic activity measured with positron emission tomography and [18F]fluoride ion in renal osteodystrophy: correlation with bone histomorphometry. AB - We evaluated the bone metabolic activity in patients with renal osteodystrophy using positron emission tomography and [18F]fluoride ion. Eight patients had secondary hyperparathyroidism (HPT), and three had low-turnover bone disease. Eleven normal subjects were also studied, and three of the eight HPT patients were reevaluated after therapy. A rate constant (K) describing the net transport of [18F] fluoride ion into a bound compartment in bone was calculated using both a three-compartment model and Patlak graphical analysis. Values of K were compared with biochemical data and with histomorphometric indices. The results indicate that K is significantly higher (P < 0.01) in HPT patients than in normal subjects and patients with low-turnover bone disease. Values of K correlated with serum alkaline phosphatase (r = 0.81) and PTH (r = 0.93) levels and with histomorphometric indices of bone formation rate (r = 0.84, P < 0.01) and eroded perimeter (r = 0.77, P < 0.05). Values of K decreased by 40 and 30%, respectively, in two patients who underwent parathyroidectomy and medical therapy. Positron emission tomography studies of bone using [18F]fluoride ion can differentiate low turnover from high turnover lesions of renal osteodystrophy and provide quantitative estimates of bone cell activity that correlate with histomorphometric data. PMID- 8408471 TI - Anabolic effects of recombinant human growth hormone in patients with wasting associated with human immunodeficiency virus infection. AB - Body wasting, characterized by disproportionate loss of body cell mass, is a feature of many chronic diseases, including infection with the human immunodeficiency virus (HIV). Therapies that merely increase energy intake do not consistently restore body cell mass in patients with the wasting syndrome. Because treatment with GH has induced nitrogen (N) retention in catabolic patients after surgery, burns, cancer, and hypocaloric feeding, we designed this study to determine whether GH could also produce an anabolic response in persons with HIV-associated weight loss. Six HIV-positive (HIV+) men with an average weight loss of 19% and six healthy weight-stable controls (HIV-) were hospitalized on a metabolic ward, where they consumed a constant metabolic diet during successive 5-day precontrol, 7-day baseline, and 7-day treatment [recombinant human GH (rhGH), 0.1 mg/kg.day] periods. The effects of rhGH on body weight, N and electrolyte excretion, energy expenditure, substrate oxidation, and integrated lipid and carbohydrate metabolism were assessed. Body weight increased promptly and progressively during treatment (2.0 +/- 0.3 and 1.6 +/- 0.2 kg in HIV+ and HIV-, respectively). Urinary N excretion decreased by 288 +/- 17 and 287 +/- 42 mmol/day in HIV+ and HIV-, respectively. Resting energy expenditure increased by 7.5% in both groups. Protein oxidation decreased, whereas lipid oxidation increased significantly. Glucose flux increased, and modest increases in fasting plasma triglyceride, glucose, and insulin levels were observed. Thus, short term rhGH treatment increased both protein anabolism and protein-sparing lipid oxidation, effects that should increase body cell mass if sustained during chronic therapy. PMID- 8408472 TI - Luteinizing hormone-releasing hormone antagonists interfere with autocrine and paracrine growth stimulation of MCF-7 mammary cancer cells by insulin-like growth factors. AB - Several studies have supported the idea that LH-releasing hormone (LHRH) antagonists have a direct effect on mammary tumor cells. In this study, we have evaluated the potential role of the insulin-like growth factors (IGFs) on the growth of MCF-7 mammary tumor cells and the effect of LHRH analogs on IGF action. The mitogenic effects of IGF-I, IGF-II, and insulin were compared. IGF-I was found to be 3 times more potent than IGF-II and 30 times more potent than insulin, suggesting that the effects of these growth factors are mediated by the IGF-I receptor. IGFs released by MCF-7 cells were measured by specific RIA after acid extraction and chromatography, so as to avoid the interference of IGF binding proteins. MCF-7 cells secreted IGF-II, but not IGF-I. Estradiol (10(-9) mol/L) stimulated IGF-II release; this release preceded the effect of estradiol on cell growth. The LHRH antagonist [Ac-D-Nal(2)1,D-Phe(4Cl)2,D-Pal(3)3,D-Cit6,D Ala10] LHR H (SB-75, CETRORELIX) inhibited basal, estrogen-induced, and IGF induced growth. Moreover, this antagonist almost completely inhibited IGF-II release from MCF-7 cells. This effect preceded the inhibition of tumor cell growth. We conclude that a LHRH antagonist can inhibit the growth of breast tumors by interfering with the autocrine action of IGF-II and by directly inhibiting the growth stimulatory effect of IGFs. PMID- 8408473 TI - Competitive polymerase chain reaction quantitation of c-erbA beta 1, c-erbA alpha 1, and c-erbA alpha 2 messenger ribonucleic acid levels in normal, heterozygous, and homozygous fibroblasts of kindred S with thyroid hormone resistance. AB - Mutations in the T3-binding domain of the thyroid hormone receptor gene c-erbA beta result in dominant negative proteins and thyroid hormone resistance syndromes. Variable clinical manifestations of resistance to thyroid hormones have been reported, including short stature and neuropsychological abnormalities. The molecular bases for heterogeneity of phenotype among and within kindreds have not been fully elucidated. Recent investigations have considered differential expression of mutant and wild-type beta 1-receptor alleles and the regulation thereof as a mechanism to explain differential sensitivity to thyroid hormones. We used reverse transcription-competitive polymerase chain reaction (PCR) to measure c-erbA beta 1, c-erbA alpha 1, and c-erbA alpha 2 mRNAs in skin fibroblasts cultured from normal subjects, heterozygotes, and a severely affected homozygous mutant of kindred S. The homozygous mutant of kindred S had severe growth and mental retardation. After reverse transcription with primers specific for each of the c-erbA mRNAs, first strand cDNAs were amplified by PCR using subtype-specific amplimers. Primer design allowed simultaneous detection of wild type and mutant messages in heterozygous fibroblasts and showed an approximately 1:1 ratio of these mRNAs in three patients. Inclusion of competitive standard cDNAs of known concentration in the PCR reactions allowed quantitation of the absolute levels of the beta 1-, alpha 1-, and alpha 2 mRNAs by comparison of products on ethidium bromide-stained agarose gels. These studies showed no effect of the presence of the mutant beta 1-allele, as fibroblast RNA from normal subjects, heterozygotes, and the homozygote gave values of 56-184, 2.8-12, and 23 40 attomol/5 micrograms total RNA for beta 1-, alpha 1-, and alpha 2 mRNAs, respectively. We conclude that these sensitive methods allow the detection of molecular species present at levels as low as 10 molecules/cell, and that this potent dominant negative receptor does not disrupt c-erbA expression at the level of mRNA. The neuropsychological sequelae of the kindred S mutation are not due to relative overexpression of the mutant allele. PMID- 8408474 TI - Influence of combined C-peptide and insulin administration on renal function and metabolic control in diabetes type 1. AB - The possible influence of C-peptide on renal function and metabolic control in patients with type 1 diabetes was examined in a double blind, randomized study. Nine patients received insulin and equimolar amounts of biosynthetic human C peptide for 1 month (group 1), and nine were given insulin only (group 2). C Peptide levels in plasma ranged from 0.3-2.6 nmol/L in group 1 during the study, whereas group 2 had undetectable levels. The urinary excretion of albumin in group 1 was 21 +/- 6 micrograms/min before the study and decreased by 40% and 55% after 2 and 4 weeks, respectively (P < 0.05). No change was seen in group 2. The glomerular filtration rate fell by 6% after 2 and 4 weeks (P < 0.05) in group 1, whereas no change was observed in group 2. Fluorescein leakage across the blood retinal barrier decreased by 30% in group 1 (P < 0.05) and was unaltered in group 2. Hemoglobin-A1c and fructosamine values decreased by 9-16% in group 1 (P < 0.05), but not in group 2. The findings suggest that administration of C-peptide plus insulin, compared to insulin alone, to type 1 diabetic patients may reduce glomerular permeability and improve metabolic control. PMID- 8408475 TI - Thyroid hormone resistance syndrome: correlation of dominant negative activity and location of mutations. AB - Generalized resistance to thyroid hormone (GRTH) is caused by multiple distinct mutations that cluster in two regions of the hormone-binding domain of the thyroid hormone beta-receptor. The mutant receptors are functionally inactive, but nevertheless inhibit normal receptor activity in a dominant negative manner. Four different GRTH mutants were studied in the transient expression assays to further examine their functional properties. The transcriptional activity of the mutant receptors correlated with their T3 binding affinities. Two distal region mutants with partial T3 binding were transcriptionally active at high T3 concentrations, but exhibited potent dominant negative activity at low T3 concentrations. Two proximal region mutants that did not bind to T3 were 5- to 10 fold less effective inhibitors of normal receptor function, indicating that dominant negative inhibition is not correlated with T3 binding activity. Each of the proximal and distal region mutants retain the ability to form heterodimers with accessory proteins and to bind to DNA effectively. Because the non-T3 binding thyroid hormone receptor isoform alpha 2 also exists in most tissues, its effects on mutant receptor function were also examined. The inhibitory activity of each of the GRTH mutants was potentiated by alpha 2 but only in the context of a positively regulated reporter gene. Thus, alpha 2 may selectively alter the degree of dominant negative activity that occurs for different target genes. We conclude that the locations of GRTH mutations may influence dominant negative activity by altering transactivating or other functions of the receptor, providing a potential basis for the phenotypic variability in different kindreds with GRTH. PMID- 8408476 TI - Growth factor production by human thyroid carcinoma cells: abundant expression of a platelet-derived growth factor-B-like protein by a human papillary carcinoma cell line. AB - As papillary thyroid carcinoma cells grow surrounding finger-like structures of stromal tissue, we postulated they may secrete a growth factor(s) for mesenchymal cells and that these would be distinct from any mitogenic factors elaborated by follicular carcinomas. Conditioned medium from both the human papillary carcinoma cell line NPA and the follicular carcinoma cell line WRO evoked a 20- to 30-fold increase in [3H]thymidine incorporation into NIH3T3 cell DNA. NPA cell growth factor activity largely eluted with 0.5 mol/L NaCl from a heparin-Sepharose column. NPA-conditioned medium competed in a platelet-derived growth factor-B (PDGF-B) RRA, and the mitogenic activity was partially blocked by an anti-PDGF-BB antibody. An immunoprecipitated PDGF-B-like protein from NPA cells was about 17 kilodaltons in a reducing gel, but, in contrast to wild-type PDGF-BB, did not change its electrophoretic mobility in an unreduced sodium dodecyl sulfate polyacrylamide gel electrophoresis. NPA cells expressed an abundant 1.4-kilobase RNA that hybridized to probes for the 5'-untranslated and amino-terminal domains of PDGF-B and was distinct from the 4.2-kilobase wild-type PDGF-B chain transcript. There were no structural changes in the PDGF-B gene, as determined by cytogenetic analysis and restriction mapping. However, the PDGF-B gene in the NPA cells was hypomethylated compared to that in normal thyroid tissue or WRO cells. In contrast, the mitogenic activity of WRO cells bound to heparin with high affinity and was blocked by a basic fibroblast growth factor (bFGF) antibody. WRO cells contained abundant bFGF mRNA. Both cell lines abundantly expressed transforming growth factor-beta mRNA. Thus, NPA and WRO cells express powerful, yet distinct, mesenchymal cell growth factors. Whereas WRO cells express abundant bFGF, NPA cells produce a novel PDGF-B-like protein, which may correspond to a mutated form of PDGF-B-chain. PMID- 8408477 TI - The relationship between body composition and plasma concentrations of exogenously administered urinary follicle stimulating hormone. AB - The aim of this study was to evaluate the plasma absorption of i.m. administered exogenous urinary follicle stimulating hormone (FSH) in relation to body fatness, leanness and cell mass. Body composition was measured by means of bioelectrical impedance. An immunoradiometric assay was used to assess the concentration of exogenous FSH in blood in patients with a pharmacologically induced hypogonadotrophism who were undergoing ovulation induction for assisted reproduction. No correlation was found between body composition and the hormone concentration in blood. According to our results, the different requirements of gonadotrophin administration typically observed in patients of different weights are probably not related to different absorption and distribution of the drug. PMID- 8408478 TI - The influence of epidermal growth factor on progesterone production by human granulosa--luteal cells in culture. AB - Epidermal growth factor (EGF) stimulates progesterone production by human granulosa--luteal cells in culture. The present study investigated some of the parameters that affect the magnitude of human granulosa--luteal cells' response to EGF. Cells from pre-ovulatory follicles obtained 36 h post-human chorionic gonadotrophin (HCG) were cultured for 12 days with or without EGF (20 ng/ml). Medium was changed every 48 h and assayed for progesterone by radioimmunoassay. DNA content of the cultured cells was determined fluorometrically. EGF was added every other day to the culture medium, starting on either day 4, 6 or 8 of culture, up to day 10, and compared with controls. When EGF was initiated on day 4, the medium had significantly higher progesterone concentration than control samples on days 6, 8, 10 and 12 of culture (P < 0.01). When EGF was withheld until day 6 or 8, progesterone concentrations were not significantly higher than control values. When EGF was added on day 4 and discontinued on day 8 or 10, progesterone concentrations were reduced significantly (P < 0.001) compared with the group where EGF was added continuously from day 4 to 10. These data suggest that: (i) human granulosa--luteal cells require the early exposure and continuous presence of EGF for the stimulatory effect on progesterone secretion, (ii) cells not exposed initially to EGF do not respond in a similar way, (iii) EGF is capable of maintaining progesterone production for a period > 12 days. Therefore, normal luteal function may require the early and continuous presence of EGF. PMID- 8408479 TI - Relaxin induces ovulations in the in-vitro perfused rat ovary. AB - The effects of human recombinant relaxin on ovulation and ovarian steroidogenesis were investigated in vitro using a perfused rat ovary model. Ovaries of equine chorionic gonadotrophin (ECG; 20 IU)-primed Sprague-Dawley rats were perfused for 21 h. Ovarian release of oestradiol and progesterone was measured during the perfusion period and the number of ovulations was estimated by counting the released oocytes at termination of the experiment. Non-treated control ovaries did not ovulate whereas addition of ovine luteinizing hormone (LH; 100 ng/ml) resulted in a mean (+/- SEM) number of ovulations of 3.0 +/- 0.8 from all treated ovaries. Relaxin (10 micrograms/ml) induced mean (+/- SEM) number of ovulations at 2.4 +/- 0.2 in all treated ovaries but did not further increase the ovulation rate when combined with LH (mean +/- SEM 3.2 +/- 0.4). All ovulated oocytes in the groups stimulated by LH showed signs of nuclear maturation (germinal vesicle breakdown) when harvested, in contrast to ovulated oocytes in the relaxin group, which were immature (presence of germinal vesicle). Progesterone and oestradiol release was significantly increased in the LH-stimulated groups but not in the group treated only with relaxin, in comparison to the untreated control group. These results demonstrate that relaxin may have a paracrine role within the ovary and may facilitate ovulation, possibly by promoting connective tissue remodelling of the follicle wall. PMID- 8408480 TI - The use of intravenous albumin in patients at high risk for severe ovarian hyperstimulation syndrome. AB - Previous experiences in subjects with other forms of third space fluid accumulation have shown that albumin is efficacious in preventing and correcting haemodynamic instability. Using a similar approach in an effort to increase the serum oncotic pressure and to reverse the leakage of fluids from the intravascular space, high risk subjects for severe ovarian hyperstimulation syndrome (SOHS) were treated with albumin. In a recent large study two high risk factors were identified, i.e. the number of oocytes and levels of serum oestradiol. Thirty-six women undergoing assisted reproductive techniques who presented both these factors, received intravenous albumin at a dose of 5% in Ringers lactate in doses of 500 ml during oocyte retrieval and 500 ml immediately thereafter in the recovery room. Daily measurements of urine output, serum and urine electrolytes, weight, abdominal girth, and haematocrit prior to and after oocyte retrieval revealed normal serum and urine electrolyte levels, and no signs of haemoconcentration. No patient in this study developed SOHS, and of course none had to be hospitalized. Vaginal ultrasound performed in the majority of the subjects revealed < or = 100 ml of peritoneal fluid 48-72 h after oocyte retrieval. The only complication from the use of intravenous albumin was the appearance of a 'flu-like condition' (low grade temperature, nausea and muscle pains) developed by 12 women between days 3 and 5 after oocyte collection. Intravenous albumin had thus prevented the development of severe ovarian hyperstimulation syndrome in an assisted reproduction programme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408481 TI - Is endometrial development in the peri-implantation period influenced by high concentrations of luteinizing hormone in the follicular phase? AB - A retrospective study was designed to examine the relationship between luteinizing hormone (LH) concentrations in the follicular phase and endometrial development in the luteal phase of natural and artificial cycles. Two types of cycle were studied: natural cycles (n = 51) in subjects with unexplained infertility were divided into two subgroups, depending on whether LH measurements in the late follicular phase were based on urine (n = 24) or plasma (n = 27) samples; and artificial cycles (n = 17), produced by the administration of a standard hormone replacement therapy, in two subgroups of women, those with premature ovarian failure (n = 10) in whom plasma LH concentrations were high, and those with unexplained infertility (n = 7) who had their hypothalamic pituitary-ovarian axis down-regulated and in whom plasma LH concentrations were low. The correlation between plasma or urine concentrations of LH in the follicular phase and the results of endometrial biopsy obtained in the luteal phase was calculated. In natural cycles, LH concentrations were similar in those with normal or retarded endometrium, and there was no significant correlation between high LH concentration and retarded endometrial development. In artificial cycles, endometrial development was not different between those with low LH concentrations (down-regulated by Zoladex) and those with high LH concentrations (premature ovarian failure). Endometrial development in the peri-implantation period does not appear to be influenced by LH concentration in the follicular phase. The reported association between high LH concentration and poor reproductive performance cannot therefore be explained by abnormal implantation consequent upon retarded endometrial development. PMID- 8408482 TI - Antisperm antibodies in cervical mucus in an unselected subfertile population. AB - To determine the incidence and the clinical significance of antisperm antibodies (ASA) in cervical secretions in an unselected subfertile population, cervical mucus samples of 192 patients with long-standing infertility were screened by means of the indirect mixed antiglobulin reaction (MAR) test allowing differentiation for immunoglobulin (Ig)A and IgG in a parallel test setting. In addition, the indirect MAR IgG test in cervical mucus was evaluated by means of IgG coated latex particles instead of sensitized erythrocytes as the indicator system. All cervical mucus samples were taken under standardized conditions. Results of ASA determination were related to microbial findings in the cervix and the outcome of sperm-mucus interaction testing in vivo and in vitro, and the subsequent fertility in a prospective study. The total incidence of cervical mucus ASA within this population was low (< 2%). A significant correlation was found between sperm antibodies of the IgG and IgA class and of IgG ASA, determined with the standard MAR and testing with latex microspheres as indicator particles. Results were not influenced by microbial colonization of the cervix and were not markedly related to lymphocytes subset testing. All ASA positive women had a negative outcome of the post-coital test, but the majority of negative post-coital tests was not caused by local mucus antibodies of the IgG and/or IgA class. In patients with positive indirect MAR testing in cervical mucus, there was no pregnancy within an observation period of > 12 months. In conclusion, the results indicate the ASA in cervical secretions are not frequent but nevertheless are a severe cause of infertility when present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408483 TI - Embryonic resistance to tumour necrosis factor-alpha mediated cytotoxicity: novel mechanism underlying maternal immunological tolerance to the fetal allograft. AB - The cytokine tumour necrosis factor-alpha (TNF alpha) has been postulated to play an essential role in the cytotoxic activity of cell-mediated immunity against allogenic or tumour cells invading the host. Several tumour cell lines, however, are resistant to TNF mediated cytotoxicity and respond paradoxically by cellular proliferation and by autocrine secretion of TNF alpha. In view of the metastatic character of the mammalian embryo, the aim of this study was to assess the potential of murine embryos to secrete TNF alpha in vitro, to express TNF receptors and to resist TNF alpha mediated cytotoxicity during their in-vitro development to the blastocyst stage. The potential of human embryos to secrete TNF alpha in vitro until the blastocyst stage was also investigated. From a total of 11 human embryos, which were allowed to proceed to blastocyst formation, seven secreted TNF alpha in the range of 2-117 pg/ml/24 h. A total of 123 C57BL/6J mouse embryos were studied of which 55% secreted TNF alpha in the range of 1.25 3.95 mg/ml/24 h. The presence of high levels of exogenous TNF alpha (10-300 IU) was not detrimental to the in-vitro development of murine embryos. Using immunohistochemical techniques, we were not able to detect the presence of type I or II TNF receptors on the surface of murine embryos. Our findings suggest that human and C57BL/6J murine embryos have the potential to secrete TNF alpha in vitro during the developmental stages leading to blastocyst formation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408484 TI - Psychological factors in the aetiology of infertility: a prospective cohort study. AB - The objectives were to identify and measure psychological factors characterizing the period following the cessation of contraception and to assess these psychological factors as predictors of the couples' fertility. A cohort of 63 couples with initially undetermined fertility status was prospectively studied, first shortly after the cessation of contraception, then 13 months later. The Child Project Questionnaire was constructed to assess psychological variations following the cessation of contraception. An Interspouse Difference Score was constructed to measure the difference between the spouses' responses. Three male and three female factors were derived from the questionnaire. The Interspouse Difference Score was significantly greater in infertile than in fertile couples. Two psychological factor scores were significantly higher in fertile subjects: the wives' level of positive expectations related to motherhood, and the husbands' quality of integration between the wish for a child and sexual relationships. Within the group of fertile couples, time to pregnancy was predicted by the husbands' above-cited factor and by the wives' frequency of thoughts and concerns related to the desired child. The results support the conclusion that in both women and men, psychological factors specifically related to the project of conceiving a child are significant predictors of the couple's fertility status. PMID- 8408485 TI - Chromosome anomalies in human oocytes in relation to age. AB - Cytogenetic analysis of oocytes remaining unfertilized after in-vitro fertilization showed that the source of data obtained could be divided into degenerating and 'healthy' oocytes. The degenerating oocytes, which showed different degrees of chromosome breakage, accounted for a quarter of the total. They were found in older patients with a mean age of 35.0 years. The healthy oocytes without chromosome breaks were mostly haploid and fell into two main groups, those with a normal MII,23,X chromosome complement, and those abnormal in which single chromatids replaced a whole chromosome. No oocytes hyperhaploid for an extra whole chromosome were found. We hypothesize that the single chromatids at second meiotic metaphase arise by precocious division of chromosome univalents at anaphase I (predivision) and that this may be the major mechanism for trisomy formation in man, rather than the non-disjunction of whole bivalents as generally assumed. PMID- 8408486 TI - Higher success rate by intracytoplasmic sperm injection than by subzonal insemination. Report of a second series of 300 consecutive treatment cycles. AB - Subzonal insemination (SUZI) and intracytoplasmic sperm injection (ICSI) were carried out in 300 treatment cycles in couples unable to be helped by conventional in-vitro fertilization treatment. More oocytes were damaged by ICSI (13.5%) than by SUZI (7.1%). The normal fertilization rate was substantially higher after ICSI (51.0%) than after SUZI (14.3%) and was related to the semen characteristics. The cleavage rate was similar for both procedures (77%). After 217 embryo transfers (72.3% of the treatment cycles) 66 pregnancies were established, i.e. pregnancy rates of 22.0% per started cycle and 30.4% per embryo transfer. So far, pregnancy loss has occurred in 27.3% of the pregnancies, nine healthy children have been born after eight deliveries and 41 clinical pregnancies are progressing uneventfully. Chorionic villus sampling or amniocentesis have been performed in 35 pregnancies and 39 normal fetal karyotypes have been obtained after cytogenetic analysis. PMID- 8408487 TI - High fertilization and implantation rates after intracytoplasmic sperm injection. AB - Previously reported better fertilization rate after intracytoplasmic single sperm injection (ICSI) than after subzonal insemination of several spermatozoa was confirmed in a controlled comparison of the two procedures in 11 patients. Intracytoplasmic sperm injection was carried out in 150 consecutive treatment cycles of 150 infertile couples, who had failed to have fertilized oocytes after standard in-vitro fertilization (IVF) procedures or who were not accepted for IVF because not enough motile spermatozoa were present in the ejaculate. A single spermatozoon was injected into the ooplasm of 1409 metaphase II oocytes. Only 117 oocytes (8.3%) were damaged by the procedure and 830 oocytes (64.2% of the successfully injected oocytes) had two distinct pronuclei the morning after the injection procedure. The fertilization rate was not influenced by semen characteristics. After 24 h of further in-vitro culture, 71.2% of these oocytes developed into embryos, which were transferred or cryopreserved. Only 15 patients did not have embryos replaced. Three-quarters of the transfers were triple-embryo transfers. High pregnancy rates were noticed since 67 pregnancies were achieved, of which 53 were clinical, i.e. a total and clinical pregnancy rate of 44.7% and 35.3% per started cycle and 49.6% and 39.2% per embryo transfer. A total of 237 supernumerary embryos were cryopreserved in 71 treatment cycles. PMID- 8408488 TI - Comparison of semen quality obtained by vibratory stimulation and masturbation. AB - Six normal males underwent both penile vibratory stimulation and masturbation in order to compare the quantity and quality of semen produced by each method. There was no significant difference in the quantity and quality of the ejaculates produced by vibratory stimulation of the penis and masturbation. In addition, biochemical analysis of the seminal fluid collected by both procedures revealed similar values between all specimens for nine organic constituents, seven inorganic constituents and seven metabolic enzymes. None of the subjects demonstrated retrograde flow of semen. These findings indicate that vibratory stimulation is a 'physiological' means of inducing ejaculation, and can produce semen of normal quality. PMID- 8408489 TI - Disturbances of sperm flagella due to failure of epididymal maturation and their possible relationship to phospholipids. AB - A recent classification of alterations of midpiece and flagellum describes the phenomenon of abnormal staining behaviour of human sperm flagella during Papanicolaou and Shorr staining. In these techniques, human sperm tails normally stain red; however, if epididymal function is disturbed, the flagella appear bluish, while they are devoid of other recognizable defects. Such spermatozoa were shown to be immotile, the motility disturbance being referred to as epididymal dysfunction. To define the physiological substrates of this descriptive phenomenon, caput spermatozoa from 10 epididymides of five patients who had undergone orchiectomy because of prostatic cancer were investigated. These spermatozoa showed severe motility disturbances, and almost all their flagella stained atypically. Attempts to stimulate such spermatozoa by pentoxifylline achieved a slight improvement in motility. However, addition of seminal plasma from fertile donors resulted in a significant improvement in motility, accompanied by an increase in the number of normally stained flagella. Even better results were achieved by incubation with liposomes made from soybean lipids, mainly phosphatidylcholine. It is concluded that poor motility and atypical staining behaviour of human caput sperm tails can at least partly be related to a lack of phospholipids in the flagellar membrane. Substitution of phospholipids may be a therapeutic approach in procedures using caput spermatozoa, although in addition to the improvement of sperm motility, the enhancement of fertilizing capacity remains to be established. PMID- 8408490 TI - Selection of acrosome-reacted human spermatozoa and their fusing ability by micro injection into the perivitelline space of hamster eggs. AB - Previously we reported the selective binding of human acrosome-reacted spermatozoa to monoclonal antibody-coated paramagnetic beads. In the present paper, the fusing ability of spermatozoa that bound to the beads was estimated by micro-injecting them into the perivitelline space of hamster oocytes. Acrosome reacted human spermatozoa were collected by monoclonal antibody-coated paramagnetic beads. Bead-bound spermatozoa were selected under a microscope and micro-injected into the perivitelline space of hamster oocytes. When the injection was begun 5 min after mixing spermatozoa with the beads, 6.6% of the injected spermatozoa fused with an oocyte. However, when the injection was begun 30 min after the mixing, no fusion occurred. Moreover, when spermatozoa were injected with the beads attached to their head, no fusion was observed. The fusing ability of spermatozoa thus prepared was compared with that of randomly chosen and injected spermatozoa. Using spermatozoa from 10 healthy volunteers, 22 independent experiments were performed. The overall fusion rate of injected, randomly selected spermatozoa was 2.8%, whilst that of bead-selected spermatozoa was 7.3%. Selection of spermatozoa by beads appeared to be beneficial to the fusion event. PMID- 8408491 TI - The effects on in-vitro fertilization of autoantibodies to spermatozoa in subfertile men. AB - Thirty-six infertile couples underwent treatment by in-vitro fertilization. In 16 couples (group 1) the male partner was positive for antisperm antibodies measured by direct mixed antiglobulin reaction, direct immunobead test, and serum and/or seminal plasma tray agglutination test. In 20 couples (group 2) the men had no such antibodies. Men with poor sperm motility were excluded. The female partners had no antisperm antibodies, and in the controls (group 2) infertility was due to a known female factor. The fertilization rate in couples without antisperm antibodies (group 2) was 72.7% compared to 50.5% when the men had antibodies. However, the pregnancy rate per embryo transfer was not significantly different in the two groups (46.1% in group 1, 33.3% in group 2). This indicates that antisperm antibodies in the male interfere with sperm--egg fusion and subsequent fertilization but once fertilization has occurred, the pregnancy rate remains the same. PMID- 8408492 TI - Exposure of human spermatozoa to the cumulus oophorus results in increased relative force as measured by a 760 nm laser optical trap. AB - Spermatozoa change their movement characteristics in response to different environmental conditions. To investigate the relative force of spermatozoa exhibiting different motility patterns, a laser optical trap was used. A laser beam at 760 nm was directed through a microscope objective and focused above the spermatozoa to create a three-dimensional optical trap. Spermatozoa were trapped at 300 mW, and laser power was reduced until spermatozoa could escape. The force generated by the flagellar movement was proportional to the laser power at which the spermatozoa escaped from the trap. Three motility patterns were studied: linear, hyperactivated, and cumulus-related. Mean escape power for spermatozoa displaying linear motility was 59.5 +/- 43 mW, for hyperactivated motility 122.3 +/- 67 mW (P < 0.0001) and for cumulus-related motility 200.6 +/- 44.2 mW (P < 0.0001). In this study, we showed that human spermatozoa generated more relative force upon exposure to the cumulus mass. The combination of small-amplitude lateral head displacement and higher relative force may produce a 'drilling' effect which is synergistic with the enzymatic digestion of the cumulus matrix during the fertilization process. PMID- 8408493 TI - Effect of aspirating needle calibre on outcome of in-vitro fertilization. AB - Thirty patients with intact ovaries undergoing ultrasound-guided transvaginal oocyte retrieval for in-vitro fertilization treatment had their ovaries randomized for follicular aspiration with small and large aspiration needles while under mild sedation. Collections using smaller diameter needles had no significant effect upon the number of oocytes collected per follicle aspirated, or on the subsequent fertilizing capacity of those oocytes. There was significant reduction in pain perceived by the patient when the smaller needle was used during the collection. PMID- 8408494 TI - Cryopreservation of mouse and human oocytes using 1,2-propanediol and the configuration of the meiotic spindle. AB - Human and mouse oocytes were cryopreserved by a slow freeze, rapid thaw method, using propanediol (PROH) as the cryoprotectant. A simulated cryopreservation was also included in the study to detect the level of damage attributable to the PROH alone. Comparison of the mouse and human oocytes cryopreserved by the same method showed opposing results, with a poor morphological survival rate of 4% observed for mouse oocytes and a subsequent normal fertilization rate of 0%. In 171 cryopreserved human oocytes a higher survival rate of 64% was achieved, and this showed more similarity to the mouse pronuclear oocytes survival of 53%. A comparison of human oocytes, cryopreserved within the cumulus and denuded of cumulus and corona prior to cryopreservation, demonstrated a higher survival rate in the denuded oocytes of 69% compared to 48%. A delay prior to cryopreservation of 1 or > or = 2 days had no effect on the immediate survival of oocytes, but culture for a further 24 h after thawing reduced survival, with the day 1 oocytes exhibiting the most dramatic reduction in survival (28%). Using a lectin binding method, abundant cortical granules were observed in all cryopreserved oocytes analysed. The meiotic spindle and chromosomes were examined in cryopreserved oocytes using fluorescence microscopy and 60% of the surviving oocytes had a normal spindle and chromosome configuration. PMID- 8408495 TI - Effect of culture systems on mouse early embryo development. AB - The amount of oxygen available to embryos in drop cultures under paraffin may be limited by the gas mixture employed, the method of equilibration or by the use of positive or negative pressure filtration. When these factors were varied, in experiments using Swiss outbred (SO) embryos, the only significant difference detected was the poorer development of embryos from the 1-cell to blastocyst stage when cultured in medium without prior equilibration. Also under these conditions, the inclusion of glucose in the medium did not increase the incidence of 2-cell block, and glutamine in the presence of glucose enhanced the development of 1-cell embryos to blastocysts. One-cell (C57BL/6 x SJL) F1 x SO embryos were successfully cultured in HEPES buffered CZB medium with added bicarbonate, under an atmosphere of air. The proportion of blastocysts formed in this medium, with a concentration of 10 mM sodium bicarbonate, was not different from that developed in CZB under 5% CO2 in air. PMID- 8408496 TI - Assessment of ultrarapid and slow freezing procedures for 1-cell and 4-cell mouse embryos. AB - Three cryopreservation procedures were assessed for the freezing of mouse 1-cell and 4-cell embryos: the slow freezing protocols with dimethylsulphoxide (DMSO, A) and propanediol (PROH, B) and the ultrarapid procedure with DMSO (C), which was described by the Monash University group [A. Trounson et al. (1987) Fertil. Steril., 48, 843-850]. The evaluation of the different procedures included survival after freezing and thawing, further development after in-vitro culture to blastocysts and the ability to implant and to form living fetuses after transfer of early blastocysts to pseudopregnant mice. In-vitro development was lower in all frozen embryos than in the unfrozen controls. Procedures A and B induced comparable results and were significantly better than ultrarapid freezing. When unfrozen blastocysts were transferred to pseudopregnant mice, 64% of them implanted in the uterine wall and 59% developed to living fetuses. For zygotes the percentages of implantation sites and living fetuses were 47 and 33% for A, 52 and 44% for B, and 29 and 17% for C, respectively. When 4-cell embryos were cryopreserved, these results were 54 and 46% for A, 60 and 53% for B and 37 and 23% for C, respectively. In the ultrarapid procedure we also looked at the influence of the freezing solution. To set up a control, mouse zygotes and 4-cell embryos were exposed to DMSO as in the ultrarapid procedure except that they were not frozen; the survival and blastocyst formation was not different from controls, but fewer living fetuses were born (31% for zygotes and 41% for 4-cell embryos versus 67% in the controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408497 TI - Unsuitability of multinucleated human blastomeres for preimplantation genetic diagnosis. AB - Multinucleated blastomeres (MNBs) were detected in 30.4% of 230 cleaved but subsequently arrested human embryos, and in 66.7% of 21 cleaved embryos rejected after preimplantation genetic diagnosis. A total of 71 MNBs from both groups of embryos was analysed by fluorescence in-situ hybridization (FISH) using simultaneous X and Y chromosome detection. The sex chromosome analyses of these MNBs suggests the existence of at least three mechanisms of MNB generation. In 95% of the embryos in which mononucleated and multinucleated blastomeres were analysed, the sex of the MNBs corresponded with the sex of the entire embryo. However, the number of sex chromosomes per MNB and their distribution in each nucleus varied greatly, indicating their unsuitability for aneuploidy diagnosis at the preimplantation stage. PMID- 8408498 TI - Pulsatility index of uterine artery in pregnant and non-pregnant women. AB - In 60 infertile women, 73 uterine arterial pulsatility indices (PI) were measured by transvaginal colour Doppler sonography. The aims were to assess uterine perfusion response in infertile women during spontaneous ovarian cycles, and to analyse the change of uterine perfusion in pregnant cases. The mean PI values (+/ SD) of 67 non-pregnant cycles were 2.30 (+/- 0.78) in the follicular phase, 2.51 (+/- 1.05) in the ovulatory phase and 2.50 (+/- 0.97) in the mid-luteal phase. The mean PI values (+/- SD) of six pregnant cycles were 1.67 (+/- 0.22), 1.89 (+/ 0.41) and 2.23 (+/- 0.69) in the corresponding phases respectively. The difference between the PI values in the follicular phase for the pregnant and non pregnant groups was significant (P < 0.05), as well as that between the follicular and mid-luteal phase for the pregnant group (P < 0.05). There was no significant difference between the PI values in the ovulatory or mid-luteal phase of the two groups. PMID- 8408499 TI - Immunolocalization of oestrogen and progesterone receptors in the human decidua in relation to prolactin production. AB - The distribution of oestrogen and progesterone receptors within the decidualized stroma of the uterus was examined in early and term human pregnancy and the results related to the effect of oestradiol and progesterone on prolactin production by decidua in vitro. In early pregnancy progesterone receptors were present in the nucleus of decidualized cells of both the capsularis and parietalis but not in glandular cells. In contrast at term progesterone receptors were located within the cytoplasm of decidual cells. Oestrogen receptors were detected only in the nucleus and were present in greater amounts in decidua capsularis than parietalis in early pregnancy, but were not detectable in term decidua. Both oestrogen and progesterone receptors were present in the nuclei of cells of arterioles within the decidua. In early pregnancy prolactin production decreased during in-vitro culture of decidua parietalis but was maintained in decidua capsularis, associated with an increase in progesterone production by the decidua capsularis. In term decidua, prolactin production in vitro was only stimulated by a combination of oestradiol and progesterone. These results suggest, firstly, that maintained decidualization and prolactin production by decidua capsularis during treatment of women in early pregnancy with the anti progestin mifepristone is not due to an absence of progesterone receptor; secondly, there is a shift in immunoreactive progesterone receptor in decidual cells from the nucleus in early pregnancy to the cytoplasm in term pregnancy. This may indicate an alteration in the action of progesterone around the time of parturition; and thirdly, in term decidua, progesterone, apparently acting through the cytoplasmic receptor, is active in increasing prolactin production in vitro only when combined with oestradiol. PMID- 8408500 TI - Cervical ripening with mifepristone (RU 486) in first trimester abortion. An electron microscope study. AB - Mifepristone (RU 486) is a steroid that binds to the progesterone receptor and acts as a progesterone antagonist. It has been used clinically to terminate an early pregnancy. In the present investigation, the effects of mifepristone on the fine structure of the cervix were evaluated in two groups of women undergoing a first trimester abortion, using a triple-blind randomized protocol. In group I (n = 18), a cervical biopsy was taken after two doses of 100 mg mifepristone or placebo orally 24 and 12 h before vacuum aspiration. In group II (n = 20), cervical biopsies were taken before and after two doses of 100 mg mifepristone or placebo orally 48 and 36 h before vacuum aspiration. Smooth muscle cells, fibroblasts and an extracellular matrix with large bundles of collagen fibrils were the main structural components in all cervical biopsies. In group I, an increased number of mast cells and signs of new blood capillary formation and collagenolysis were observed in several of the biopsies after mifepristone treatment (67% correctly classified). In group II, the effects were less evident and in most of the cases (70%) no definite differences were noted between the biopsies taken before and after mifepristone treatment. Nevertheless, mifepristone treatment had induced softening in the cervical tissue, as judged by the decrease in resistance to mechanical dilatation. PMID- 8408501 TI - Induced regeneration of endometrium following curettage for abortion: a comparative study. AB - A significant increase in endometrial thickness and volume was observed in 30 patients given oestrogen and progestin supplementation following curettage for first trimester abortions, compared with 30 women who received no treatment. This indicates an enhanced regeneration of the endometrium following treatment. The ability to induce this response, creating a space between the intra-cavity surface area a short time after abortion, may theoretically be suggested as preventative treatment to reduce the risk of intrauterine adhesions. PMID- 8408502 TI - High fecundity rates in donor oocyte recipients and in-vitro fertilization surrogates using parenteral oestradiol valerate. AB - Ovum donation and in-vitro fertilization (IVF) surrogacy can help couples with difficult infertility problems achieve pregnancy. Most centres using oral oestrogens and oestradiol patches report pregnancy rates in the range of 30% per cycle. Parenteral oestradiol valerate has pharmacological properties that make it an attractive option for preparing the endometrium in the recipients undergoing these procedures. When the egg providers were under age 35 years, and using oestradiol valerate in the recipients, we achieved a 61% clinical pregnancy rate in 62 cycles. These improved results suggest that parenteral oestradiol valerate should be used to prepare the endometrium in recipients, and that the hormonal milieu of the endometrium plays an important role in the higher implantation rates obtainable in ovum donor and IVF surrogate cycles. PMID- 8408503 TI - Inadvertent gonadotrophin-releasing hormone agonist (GnRHa) administration in the luteal phase may improve fecundity in in-vitro fertilization patients. AB - Spontaneous pregnancies associated with inadvertent periconceptional administration of a gonadotrophin-releasing hormone agonist (GnRHa) occur in approximately 1% of in-vitro fertilization (IVF) cycles. The two main issues to be considered in these circumstances are the luteolytic effect of the agonist and embryotoxicity. In addition, some authors have suggested a higher incidence of ectopic implantations. In view of these concerns, we report on 15 patients who conceived during pituitary desensitization with a GnRHa in the luteal phase of the menstrual cycle, and review the literature on the subject. A detailed analysis of the data available so far, which include 59 pregnancies exposed to GnRHa, shows that: (i) there is no clinical evidence for impaired luteal function, and hormonal supplementation does not improve pregnancy outcome; (ii) with only two cases of reported minor malformations among 37 deliveries, both having a genetic component, there is no evidence of teratogenic effects; and (iii) ectopic implantations in these circumstances are related to tubal disease but not to the drug. Considering the long history of infertility in these patients who had previously been treated unsuccessfully by different therapeutic modalities, it is likely that the occurrence of those pregnancies is not merely coincidental and that GnRHa might have a positive role in fecundity. The improved fecundity may be explained by the mechanisms of luteinizing hormone action in the corpus luteum. PMID- 8408504 TI - Recommendations of the French College of Obstetricians and Gynaecologists for the diagnosis, treatment, cost and results of the treatment of infertility in health services. PMID- 8408505 TI - Intracytoplasmic sperm injections (ICSI) and human fertilization: does calcium hold the key to success? PMID- 8408506 TI - Follicular stimulation and ovarian cancer? PMID- 8408507 TI - IFFS expert group report on the Whittemore study related to the risk of ovarian cancer associated with the use of infertility agents. PMID- 8408508 TI - Fertility drugs and risk of ovarian cancer. PMID- 8408509 TI - Male infertility: is its rising incidence due to better methods of detection or an increasing frequency? PMID- 8408510 TI - The search for improved in-vitro systems should not be ignored: embryo co-culture may be one of them. PMID- 8408511 TI - Transplantation of frozen-thawed mouse primordial follicles. AB - Primordial follicles were isolated from juvenile mouse ovaries and cryopreserved by slow freezing with dimethylsulphoxide as the cryoprotectant. After thawing, approximately 80% of the oocytes and 65% of the somatic cells excluded Trypan Blue dye, indicating that cell membranes were still intact. Frozen-thawed cells were suspended in plasma clots and transplanted to the ovarian bursas of host animals that had been sterilized by oophorectomy. The grafts of frozen-thawed cells reorganized into morphologically distinguishable ovaries which produced signs of oestrogenic activity. After natural mating, host females produced normal offspring that were demonstrated by genetic markers to be derived from the transplanted frozen-thawed primordial follicles. PMID- 8408512 TI - Autonomic nervous modulation and effects of a prostaglandin synthase inhibitor on human cervical secretion. AB - Modulation of cervical secretion at ovulation time was studied in 10 women with regular menstruations. In an in-vivo model with repeated collection of mucus samples during three 90-min periods, the amounts of mucus in a control cycle and in three experimental cycles were compared. Drugs interacting with the autonomic nervous system and a prostaglandin synthase inhibitor were administered at time of ovulation. The cholinomimetic drug carbacholine significantly increased cervical secretion, while the anticholinergic drug butylscopolamine markedly inhibited this secretion. A long-lasting decrease in secretion was seen after administration of the prostaglandin synthase inhibitor diclofenac. Beside regulation of cervical secretion by the ovarian hormones, these results suggest an autonomic nervous modulation of cervical secretion, and in addition an impact on cervical by a prostaglandin synthase inhibitor. The effects on fertility regulation in the female are discussed. PMID- 8408513 TI - Insulin hypersecretion together with high luteinizing hormone concentration augments androgen secretion in oral glucose tolerance test in women with polycystic ovarian disease. AB - Female hyperandrogenism is often associated with hyperinsulinaemia and insulin resistance. We evaluated the hormone responses in an oral glucose tolerance test to investigate the interactions of gonadotrophins, insulin, C-peptide and androgens in women with polycystic ovarian disease (PCOD). In 28 patients with ultrasonographically diagnosed PCOD, hyperinsulinaemia and insulin resistance were mainly associated with obesity. Both basal and cumulative sum of insulin to C-peptide ratios were high in obese subjects, suggesting decreasing hepatic removal of insulin caused by obesity. Nevertheless, in some lean PCOD women, despite normal fasting insulin concentrations, insulin hypersecretion existed. The mean concentration of testosterone decreased significantly during the oral glucose tolerance test both in PCOD and control women, and of androstenedione in the PCOD patients only. However, an increase in androgen responses was found in a subgroup of PCOD patients, who had both elevated luteinizing hormone (LH) concentrations and hyperinsulinaemic response to oral glucose. In the remaining PCOD patients an inverse correlation between LH and insulin was found. The patients with hyperinsulinaemia together with LH hypersecretion may represent a subgroup of PCOD with deranged regulation of androgen secretion. PMID- 8408514 TI - Repeated aspiration of ovarian follicles and early corpus luteum cysts in an in vitro fertilization programme reduces the risk of ovarian hyperstimulation syndrome in high responders. AB - A retrospective analysis of ovarian hyperstimulation syndrome in high responders undergoing in-vitro fertilization (IVF) is presented. High responders were defined as having > 20 follicles and serum oestradiol > 3000 pg/ml after treatment with human menopausal gonadotrophin. Of the initial 30 IVF cycles in high responders, 23 developed a moderate-to-severe ovarian hyperstimulation syndrome (76.7%). Subsequently, 15 other IVF cycles in high responders were combined with a repeated aspiration of ovarian follicles and corpus luteum cysts just prior to embryo transfer. Only three patients (20%) developed a moderate ovarian hyperstimulation syndrome (P = 0.0004). We conclude that repeated follicular aspiration is safe and results in a significant reduction in the incidence and severity of this condition in high responders undergoing IVF. PMID- 8408515 TI - The impact of cigarette smoking on the plasma concentrations of gonadotrophins, ovarian steroids and androgens and upon the metabolism of oestrogens in the human female. AB - This study compares the plasma gonadotrophin, oestradiol, and androgen and salivary progesterone concentrations in a single menstrual cycle between 25 normal pre-menopausal women who smoke cigarettes and 21 who are non-smokers. The effect of smoking on luteinizing hormone (LH) pulsatility and the urinary excretion of oestrogens is also described. Cigarette smoking did not consistently suppress LH pulsatility. There was no significant difference in the length of either the follicular or luteal phases. There were no significant differences in the mean plasma oestradiol concentrations in the follicular phase in smokers compared to non-smokers. There were no significant differences in the mean salivary progesterone concentration in the luteal phase in smokers compared to non-smokers. There was no significant difference in plasma concentrations of testosterone, androstenedione and dehydroepiandrosterone sulphate. There was also no significant difference between the urinary concentrations of oestradiol, oestrone or oestriol. We have been unable to demonstrate a detrimental effect of cigarette smoking on any of the important endocrine characteristics of the menstrual cycle, and we conclude that these data suggest that the anti oestrogenic effect of smoking does not work through alterations in pituitary or follicular endocrine function or in alterations in the metabolism of oestrogens. PMID- 8408516 TI - Insulin actions on ovarian steroidogenesis are not modulated by metformin. AB - Metformin, an agent used in treatment of non-insulin-dependent diabetes mellitus, is believed to act by potentiating the effects of insulin on glucose metabolism. This study was designed to determine whether metformin affects the actions of insulin on ovarian steroidogenic capability. Rat thecal-interstitial (T-I) and granulosa (G) cells were cultured in chemically defined media for 144 h with or without gonadotrophins [luteinizing hormone (LH) at 100 ng/ml or follicle stimulating hormone (FSH) at 100 ng/ml], insulin (1 microgram/ml) and/or metformin (1 and 5 micrograms/ml). Production of testosterone and progesterone by T-I cells, and 17 beta-oestradiol and progesterone by G cells were assessed. Insulin potentiated LH-dependent stimulation of testosterone production by T-I cells and FSH-dependent stimulation of 17 beta-oestradiol production by G cells, but did not significantly affect progesterone production by T-I cells or G cells in the presence or absence of gonadotrophins. Metformin did not affect any of the actions of insulin on steroidogenesis. These results suggest that insulin may modulate ovarian steroidogenesis via a pathway separate from that modulating glucose metabolism. Actions of insulin on steroidogenesis are selective with regard to stimulation of specific aspects of steroidogenesis and do not simply amplify gonadotrophin effects. PMID- 8408517 TI - Evidence for the in-vitro de-novo synthesis of immunoglobulin and a previously undescribed 17 kDa protein (TEP-2) by the mucosa of the fallopian tube. AB - De-novo synthesis and secretion of proteins by short-term explants of matched Fallopian tube mucosa and endometrium were studied using radiolabelled L [35S]methionine and [3H]glucosamine. To compare directly each anatomical site of the Fallopian tube and endometrium from the same source, newly synthesized proteins were separated on one-dimensional polyacrylamide gel electrophoresis and examined by auto-radiography. De-novo synthesis of two protein bands provisionally designated as tubal epithelial protein 1 (TEP-1) and tubal epithelial protein 2 (TEP-2), was observed in explants of the Fallopian tube mucosa obtained from each anatomical site throughout the ovarian cycle (n = 20). TEP-2 was not apparent in tubal mucosa obtained from post-menopausal women (n = 5). De-novo synthesis of TEP-1 and TEP-2 was not apparent in autoradiographs of radiolabelled proteins from short-term explants of endometrium. From the autoradiographs the molecular mass of TEP-1 and TEP-2 was calculated to be 25 kDa and 17 kDa, respectively. Incorporation of glucosamine into newly synthesized protein occurred in TEP-2 but not TEP-1. TEP-1 was observed to be immunochemically identical to immunoglobulin kappa light chains. PMID- 8408518 TI - Treatment of male infertility: is it effective? Review and meta-analyses of published randomized controlled trials. AB - There are few widely accepted treatments for male subfertility. Controlled trials are necessary to evaluate different subfertility treatments because pregnancies and spontaneous improvements in semen parameters occur without treatment. Properly randomized trials can find out whether pregnancy rates are higher or lower than expected after treatment. Quantitative meta-analysis improves precision where individual trials are not powerful enough to demonstrate moderate treatment effects. Only 174 published randomized treatment trials for male subfertility were found after extensive literature review, both by computerized search and hand searches of 41 journals. In 72 of the studies, pregnancy was an outcome measure and these are analysed and discussed. The quality of most trials is poor. Meta-analysis was possible on seven themes, but the results of this exercise appear to be unduly influenced by trials of poor quality. Few conventional treatments (i.e. those not involving assisted conception techniques) result in improved fertility rates. PMID- 8408519 TI - Gamete intra-fallopian transfer: outcome following the elective or non-elective replacement of two, three or four oocytes. AB - The outcome of 807 gamete intra-Fallopian transfer (GIFT) cycles following the elective or non-elective transfer of two, three or four oocytes has been retrospectively studied. Electively replacing either three or two oocytes did not reduce the clinical or ongoing pregnancy rate when compared with replacing four oocytes. The incidence of high-order multiple gestation (triplet or more) was significantly reduced by replacing fewer oocytes, but the occurrence of twin pregnancy was not altered. Lower pregnancy rates were found when the number of oocytes available for replacement was limited and non-elective replacement was performed. It is suggested, therefore, that a higher number of oocytes available may allow selection of higher quality oocytes for transfer. We conclude that the overall expectation of pregnancy from the GIFT procedure is high (30-40%) and the number of oocytes replaced should be two in order to minimize the risk of high order multiple pregnancies. It is not clear whether increasing the number of oocytes transferred will benefit subjects who failed to become pregnant previously with GIFT, but limited data suggest that transferring large numbers of oocytes to women > 40 years does not improve the expectation of pregnancy. PMID- 8408520 TI - Osmo-sensitivity of the human sperm acrosome reaction. AB - The osmo-sensitivity of the human sperm acrosome reaction was investigated by determining the effect of hyper- and hypo-osmolal conditions on the ionophore A23187- and dbcAMP-induced reaction of both capacitated and non-capacitated spermatozoa. Hyper-osmolal conditions inhibited the agonist-induced reactions of both types of spermatozoa. Hypo-osmolal conditions caused a spontaneous loss of acrosomes from capacitated but not from non-capacitated spermatozoa. The loss of acrosomes under hypo-osmolal conditions was further enhanced by dbcAMP but not by ionophore A23187. Although significant decreases in sperm viability were occasionally observed at the high and low osmolalities, these decreases were not consistent and could not account for the observed loss of acrosomes. It is concluded that the human sperm acrosome reaction is osmo-sensitive. The acrosome reaction stimulated by ionophore A23187 (raises intracellular Ca2+) and dbcAMP (activates protein kinase A which causes protein phosphorylation) appears to involve water entry downstream from the action of these agonists. Preincubation in albumin (capacitation) causes human spermatozoa to lose their acrosomes under hypo-osmolal conditions. Finally, capacitation is not an essential prerequisite to the acrosome reaction as long as agonists are used that by-pass certain membrane-related events. PMID- 8408521 TI - The ability of the hemizona assay to predict human fertilization in different and consecutive in-vitro fertilization cycles. AB - The objective of this prospective study was to examine the ability of the hemizona assay (HZA) to predict fertilization outcome of mature, pre-ovulatory oocytes under in-vitro fertilization (IVF) conditions. Since a large number of patients were evaluated over a long period, the power of the HZA to prognosticate fertilization results in the same and subsequent (consecutive) IVF cycles of those same patients was assessed. For IVF, only metaphase II oocytes were used. For the HZA, both fresh oocytes donated by patients at the time of IVF and oocytes recovered from surgically removed ovarian tissue (and salt-stored) were used, and bisected by micromanipulation techniques. Matching hemizonae were co incubated either with spermatozoa from the patient (test) or from a fertile man (control) for 4 h. The number of spermatozoa tightly bound to the zona was counted. Patients (n = 112) were divided into two groups based on HZA results (expressed as HZA index or HZI): HZI > or = 30% (n = 72) and < 30% (n = 40). The patients with HZI < 30% had significantly lower fertilization rates in both the HZA-IVF cycle and in subsequent cycles compared to patients with HZI > or = 30% (P < 0.03). Linear discriminant analysis indicated the HZA to have a sensitivity of 84%, and positive and negative predictive values of 85 and 70% respectively, for prediction of fertilization outcome in a total of 233 cycles. It was concluded that the HZA is a good predictor of fertilization rate in vitro, and can be used in the IVF setting to supply additional clinical information in male factor patients. PMID- 8408522 TI - Subzonal sperm insertion with aged human oocytes from failed in-vitro fertilization attempts: fertilization results and some applications. AB - This study was undertaken to assess the potential of aged human oocytes from failed in-vitro fertilization attempts as a model for the study of fertilization events after subzonal sperm insertion (SUZI). Criteria of aged oocyte suitability for this purpose were (i) the absence of nuclei, (ii) the presence of a polar body, (iii) the absence of cell division or fragmentation, (iv) marked ooplasmic contraction in hyperosmotic medium, and (v) rapid ooplasmic relaxation after returning into normo-osmotic medium following the micromanipulation. Micro injection techniques were essentially the same as for SUZI with fresh oocytes. Oocytes that fused with the micro-injected spermatozoa developed pronuclei of typical internal structure. However, the number of pronuclei was often higher than that theoretically expected if each sperm nucleus incorporated into the oocyte gave rise to a single pronucleus. Thus, the use of aged oocytes implies the need for a specific method to assess the frequency of fusion in sperm samples examined. The results suggest that the method described here can be applied in a preliminary diagnostic test before a therapeutic SUZI attempt and in studies aimed at the optimization of sperm treatment protocols to increase the fusion capacity of subzonally inserted spermatozoa. PMID- 8408523 TI - Effects of multiple ejaculations after extended periods of sexual abstinence on total, motile and normal sperm numbers, as well as accessory gland secretions, from healthy normal and oligozoospermic men. AB - Four healthy donors provided three ejaculates on one day to deplete their sperm reserves and also after abstention periods of 1, 2, 4, 7, 10 and 14 days. The percentage of spermatozoa that were normally formed, progressively motile and which swelled in hypotonic medium remained unchanged throughout this period and the increase in total numbers of viable spermatozoa for 10 days prompted a comparison of seven donors and six oligozoospermic men. Multiple ejaculations after 10 days of abstinence for the patients with low normal, motile sperm numbers increased the total number, number of normal and number of motile spermatozoa by a mean of 230, 370 and 520%, respectively. PMID- 8408524 TI - The multicentre transcervical balloon tuboplasty study: conclusions and comparison to alternative technologies. AB - Transvaginal tubal catheterization procedures have been suggested as an alternative to microsurgery and in-vitro fertilization (IVF) in the treatment of women with proximal tubal occlusion. A transcervical balloon tuboplasty (TBT) catheter was specifically developed and tested in a prospective multicentre trial. A total of 151 women with confirmed bilateral or unilateral tubal occlusion were studied. The primary study population included 106 women who, after exclusion of patients for protocol violations, represented those females who were treated for complete tubal occlusion with TBT. TBT is an ambulatory, minimally invasive catheter procedure, performed under paracervical block or mild sedation, which utilizes a co-axial balloon catheter under fluoroscopic guidance. Re-canalization, pregnancy and reocclusion rates following the procedure were documented. A total of 28 patients demonstrating uni- or bilateral tubal patency after either hysterosalpingography and/or selective salpingography represented the control population. TBT established tubal patency of at least one Fallopian tube in 95/106 patients (90%) and in 167/205 obstructed oviducts (82%). Clinical pregnancies occurred in 37/106 females (35%), with a life table adjusted rate of 37%. Patients without distal disease had significantly higher pregnancy rates than those with bipolar tubal disease (49% versus 12%, life table adjusted rate; P = 0.0002) but pregnancy rates were independent of underlying aetiology for tubal disease. Pregnancy rates in control patients who did not reach TBT because of tubal patency after hysterosalpingography and/or selective salpingography were significantly lower than in those successful treated with TBT (P = 0.027), and occurred only for four cycles after hysterosalpingography and with approximately a 1 year delay after selective salpingography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408525 TI - Vaginal squamous cells in follicular aspirates following transvaginal puncture. AB - Squamous cells are common findings on cytological examination of fluid obtained following transvaginal aspiration of simple ovarian cysts. This prospective study confirms the hypothesis that these cells are commonly introduced as contaminants during puncture of the vaginal wall. The occasional finding of bacteria also suggests a possible mechanism for post-operative pelvic infection. PMID- 8408526 TI - The role of calcium in mammalian oocyte maturation and egg activation. AB - The maturation of the immature oocyte and the fertilization of a mature egg are two absolute prerequisites for mammalian embryo development. There is increasing evidence in mammals that both oocyte maturation and egg activation at fertilization are controlled by changes in intracellular free Ca2+ levels. The role of Ca2+ changes at fertilization is clear in that they are both required and sufficient for egg activation. However, it is not established how the sperm causes Ca2+ changes in eggs at fertilization, nor how different patterns of Ca2+ change affect embryo development. The role of Ca2+ in triggering oocyte maturation is less clear, although preventing intracellular Ca2+ changes can inhibit meiotic maturation at specific stages. Studies on how Ca2+ regulates meiosis and fertilization in mammals may provide new insights into the causes of failed fertilization in human IVF procedures. PMID- 8408527 TI - Ovarian abscess following puncture of an endometrioma during ultrasound-guided oocyte retrieval. AB - A 34-year-old white female patient with 11 years of primary infertility due to endometriosis underwent ultrasound-guided oocyte retrieval for in-vitro fertilization. A right ovarian endometrioma was aspirated during oocyte retrieval. Two weeks after follicular aspiration, the patient developed acute abdominal pain and operative laparoscopy revealed a ruptured right ovarian abscess. She responded to laparoscopic drainage and intravenous antibiotics. The patient conceived during that cycle and has a single ongoing pregnancy. PMID- 8408528 TI - Analysis of the risk factors with regard to the occurrence of ectopic pregnancy after medically assisted procreation. AB - In a retrospective study (1985-1989) based on data from the Centre for Reproductive Medicine in Brussels, a total of 23 ectopic pregnancies (2.24%) occurred after 3800 embryo, zygote or gamete transfers. This number was low compared with the data published elsewhere. Tubal damage was a major risk factor towards developing an ectopic pregnancy after in-vitro fertilization and embryo transfer. The number of ectopic pregnancies after the association of clomiphene citrate and human menopausal gonadotrophin (HMG) was significantly higher in patients with tubal (7.8%) and non-tubal indications (2.1%) compared with those stimulated with gonadotrophin-releasing hormone (GnRH) and HMG (2.18% and 0.84%, respectively). The number of replaced embryos was not associated with the rate of ectopic pregnancy and neither did transfer technique (intra-uterine or intra Fallopian transfer) influence the ectopic pregnancy rate. PMID- 8408529 TI - The effect of RU486 on progesterone and oestrogen receptor concentration in human decidua on early pregnancy. AB - Concentrations of progesterone receptor (PR) and oestrogen receptor (ER) were measured by radioligand assay in decidual tissue of women undergoing termination of early pregnancy (amenorrhoea up to 49 days). Pregnancies were terminated by vacuum aspiration at 12 or 36 h after oral administration of placebo or antiprogestin RU486 in different doses. Treatment with RU486 decreased decidual PR content, the effect being observed at 12 h as well as at 36 h after 600 mg RU486 and at 36 h after 3 x 25 mg RU486 given at 12 h intervals. PR concentration 12 h after a single dose of 25 mg RU486 was not affected. ER content was unchanged at 12 h after RU486 but increased 36 h after 600 mg and 3 x 25 mg RU486. Our data suggest that apart from blocking progesterone action, RU486 may exert its abortifacient effect through decreasing the PR concentration. The simultaneous decrease of PR concentration and an increase of ER concentration changes the balance between them in favour of ER, which might also play a role in the abortifacient effect of RU486. PMID- 8408530 TI - Adverse effect of a homogeneous hyperechogenic endometrial sonographic pattern, despite adequate endometrial thickness on pregnancy rates following in-vitro fertilization. AB - We have previously presented data to show that in patients who had in-vitro fertilization (IVF)-embryo transfer using ovarian stimulation involving the luteal phase leuprolide acetate--human menopausal gonadotrophin (HMG) regimen, poor pregnancy results ensued if either the endometrial thickness was < 10 mm or a homogeneous hyperechogenic sonographic pattern was present immediately prior to taking a human chorionic gonadotrophin (HCG) injection. There were only 15 cases with this hyperechogenic type endometrium (and no pregnancies). The purpose of the present study was to evaluate the influence of a hyperechogenic endometrium when the endometrial thickness was > or = 10 mm, in a more extensive series, in women having IVF-embryo transfer using the same ovarian stimulation regimen. A total of 273 consecutive cycles, where endometrial thickness was > or = 10 mm, were evaluated (not including the 85 cycles previously reported). Of 22 patients with the hyperechogenic pattern, one achieved a chemical pregnancy (beta-HCG > 500 mIU/ml) and none achieved clinical pregnancies (ultrasound confirmation). In contrast, 67 of 251 (26.7%) patients conceived with other echo patterns (chi 2 analysis = 5.9, df = 1, P = 0.01). These data thus confirm, in a larger series, the negative influence of this type of echo pattern on subsequent pregnancy rates following the luteal phase leuprolide acetate--HMG ovarian stimulation regimen. PMID- 8408531 TI - The increased risk of complication observed in singleton pregnancies resulting from in-vitro fertilization (IVF) does not seem to be related to the IVF method itself. AB - Singleton pregnancies resulting from in-vitro fertilization (IVF) seem to have an increased risk of obstetric and paediatric complications. In a retrospective study we compared, during the same period, 162 IVF singleton pregnancies with 263 pregnancies resulting from stimulated cycles (without IVF) and with 5096 natural pregnancies. No significant difference was found between the first two groups concerning complications (i.e. prematurity, low birthweight, small-for gestational-age and perinatal mortality). The results indicate that an adverse outcome is more common for pregnancies after ovarian stimulation (with or without IVF) compared to natural pregnancies. Therefore the increased risk does not appear to be linked to the IVF method itself but rather to a common factor in these two populations, i.e. population characteristics, underlying infertile status and/or ovarian stimulation. PMID- 8408532 TI - Laparoscopy: a dispensable tool in the diagnosis of ectopic pregnancy? AB - Laparoscopy is regarded as the final decisive diagnostic test in suspected ectopic pregnancy. The new non-invasive diagnostic methods of transvaginal sonography and serum human chorionic gonadotrophin (HCG) monitoring now challenge this pivotal role of laparoscopy. In this prospective study the diagnostic value of an algorithm, combining transvaginal sonography with an HCG cut-off level between 1000 and 1500 IU/l (IRP) was tested in 208 consecutive women at risk for ectopic pregnancy. Three diagnostic categories are designated by the algorithm: intra-uterine pregnancy (n = 73), ectopic pregnancy (n = 89), and trophoblast in regression (n = 46). The latter category represents patients in whom no pregnancy could be located by transvaginal sonography, with an initial HCG concentration < 1500 IU/l, declining during follow-up. The algorithm has a sensitivity of 0.97, a specificity of 0.95, a likelihood ratio for a positive test of 19.4, and a likelihood ratio for a negative test of 0.03. The described diagnostic strategy thus proved extremely reliable in the safe management of patients at risk for ectopic pregnancy, and renders laparoscopy obsolete. PMID- 8408533 TI - Transvaginal sonography and human chorionic gonadotrophin measurements in suspected ectopic pregnancy: a detailed analysis of a diagnostic approach. AB - In this prospective study among 208 high-risk patients with suspected ectopic pregnancy, the diagnostic value of transvaginal sonography and serum human chorionic gonadotrophin (HCG) measurements were analysed in detail. The absence of an intra-uterine gestational sac obviously was the most constant sonographic finding among patients with ectopic pregnancy (n = 89), with a very high sensitivity (0.99) but a low specificity (0.41). The application of different HCG cut-off levels improved specificity to 1.00 for values exceeding 4500 IU/l. Clinical utility obviously decreased, as many patients presented with HCG values well below this level. The additional effect of adnexal findings was analysed. Sonographic identification of an ectopic pregnancy was very specific (0.99) but had low sensitivity (0.56) because many ectopics were not detected. The additional effect of HCG values on these results was minor. The low HCG cut-off levels advocated in recent studies are questioned by the results of our analysis: whereas the combined use of sonography and HCG measurements is shown to be of great benefit, the limitations are also documented, underlining the need for re evaluation at intervals of patients with low HCG values. The question of which cut-off level should be used in practice, however, hinges on a difficult choice between a certain specificity and clinical utility. PMID- 8408534 TI - Endometrial ultrasonography as a predictor of pregnancy in an in-vitro fertilization programme. AB - The endometrial pattern and thickness were analysed by ultrasonography in 139 cycles stimulated for in-vitro fertilization (IVF) on the day of administration of human chorionic gonadotrophin (HCG). A semi-programmed schedule based on the pill + clomiphene citrate + human menopausal gonadotrophin (HMG) was used in all cycles. On the day of HCG administration, endometrial pattern and thickness were assessed with an Ultramark 4 (ATL) ultrasound equipped with a 5 MHz vaginal probe. Endometrial pattern I (a 'triple-line' multilayer) was observed in a total of 105 cycles (76%), and pattern II (fully homogeneous and hyperechogenic in relation to myometrial tissue) in 34 (24%). The incidence of clinical pregnancy did not differ (P = 0.52) between the groups with endometrial patterns I (23.8%) and II (29.4%). Endometrial thickness on the day of HCG administration in the group with pattern I (8.4 +/- 1.9 mm) was similar (P = 0.96) to that observed in the group with pattern II (8.4 +/- 2.0 mm). In addition, the endometrial thickness of the patients who became pregnant (8.0 +/- 1.7 mm) did not differ (P = 0.15) from that of women who did not achieve pregnancy (8.6 +/- 2.0 mm). The conclusion from the present data is that ultrasonographic analysis of endometrial thickness and refringency on the day of HCG administration had no predictive value for conception in IVF cycles. PMID- 8408535 TI - A genetically defined animal model of anembryonic pregnancy. AB - Mutant mouse conceptuses, homozygous for a defect in the gene encoding the glycolytic enzyme, glucose phosphate isomerase (GPI) died in utero by 9 1/2 days post coitum. By this stage there was no normal embryo, but trophoblast and some extra-embryonic membranes usually survived. The morphology of these genetically determined, anembryonic conceptuses was similar to some of the sporadic cases of anembryonic conceptuses that occurred in control crosses. These similarities suggest that an understanding of the cause of death of the homozygous mutant embryos might shed light on the aetiology of some of the cases of sporadic anembryonic conceptuses in both mouse and man. At implantation, the conceptus depends on anaerobic glycolysis for its energy production and any shortage of substrates would compromise development. It is argued that, in the absence of efficient glycolysis, a homozygous null embryo would be unable to produce sufficient energy to develop normally beyond the egg cylinder stage. However, the outer part of the conceptus might survive if nutrients and oxygen were available from the surrounding maternal tissue to produce energy by the aerobic tricarboxylic acid cycle. The homozygous GPI-null conceptuses may provide a useful animal model for some types of human anembryonic pregnancy. PMID- 8408536 TI - The influence of the number of embryos transferred in 1060 in-vitro fertilization pregnancies on miscarriage rates and pregnancy outcome. AB - To assess the incidence of miscarriage, multiple pregnancy and outcome of pregnancy in relation to the number of embryos transferred during in-vitro fertilization (IVF), an analysis was performed of 1060 pregnancies conceived in a tertiary-referral IVF clinic. There was no difference in the miscarriage rate after transfer of one or two embryos (37.7% and 34.6%), or after three or four embryos (22.5% and 25.2%). The miscarriage rate was, however, higher when one or two embryos were transferred compared with three (P < 0.01) or four embryos (P < 0.02). Of the 724 ongoing pregnancies, 524 (72.3%) were singleton, 164 (22.7%) twin, 33 (4.6%) triplet and three (0.4%) quadruplet. The mean (+/- SD) ages of women with singleton, twin, triplet and quadruplet pregnancies were 32.5 (+/- 3.8), 32.0 (+/- 3.5), 29.76 (+/- 4.3) and 29.67 (+/- 2.5) years respectively. The mean age of women with singleton and twin pregnancies was similar and both were greater than that of triplet pregnancies (P < 0.007). The overall perinatal mortality rate (PNMR) was 39.7/1000. The PNMR for singletons was 17.2/1000, for twins 80.0/1000 and for triplets 30.6/1000. All of the babies from the three quadruplet pregnancies survived. There were more babies lost in the twin pregnancies than any other group, although this only reached significance for singletons versus twins (P < 0.00005). We conclude that the incidence of miscarriage is increased in women in whom one or two embryos are transferred. Multiple pregnancies are more likely to occur in younger women and are associated with a significantly higher rate of perinatal mortality. PMID- 8408537 TI - Severe ovarian hyperstimulation syndrome, selective embryo reduction and heterotopic pregnancy. AB - Ovarian hyperstimulation syndrome is common (21.4%) in patients with polycystic ovarian disease, treated by gonadotrophins. It is much frequent (50%) in conceptual cycles. We report a case associated with a quadruplet pregnancy that underwent selective embryo reduction at 8 weeks' gestation to a twin pregnancy and was subsequently found to have an unruptured ectopic pregnancy at 11 weeks' gestation. After laparotomy and partial salpingectomy a successful twin pregnancy ensued. PMID- 8408538 TI - An evaluation of counselling for couples undergoing treatment for in-vitro fertilization. AB - Fertility difficulties, along with their investigation and treatment, are widely believed to cause significant psychological problems. This study was designed to investigate the efficacy of a non-directive counselling intervention with couples undertaking their first cycle of in-vitro fertilization treatment. Couples were randomly assigned to either a control group, given information about the treatment programme, or to an experimental group, given the same information plus three sessions of counselling before, during and on conclusion of the first treatment cycle. Psychological assessments were made at three points in the treatment process. Ratings were also obtained from the couples on the stress engendered by different parts of the treatment, the effects on their relationship and satisfaction with counselling. The results showed the patients to be generally well adjusted and anxiety levels dropped over the course of treatment. Counselling compared to information alone did not lead to any enhanced reduction in levels of anxiety or depression. The implications of the findings for service provision are discussed. PMID- 8408539 TI - Endometrial thickness and echo patterns. PMID- 8408540 TI - Successful fertilization by testicular spermatozoa in an in-vitro fertilization programme. PMID- 8408541 TI - Construction of a bioluminescent mycobacterium and its use for assay of antimycobacterial agents. AB - To show, as a model system, that mycobacteria can express heterologous luciferase genes and that bioluminescence can be a rapid method of measuring antimycobacterial activity, a bioluminescent form of Mycobacterium smegmatis was made by transformation with a Mycobacterium-Escherichia coli shuttle vector containing the luxAB genes from Vibrio harveyi. The antimycobacterial effects of antibiotics and biocides could be assayed in real time by using bioluminescent M. smegmatis. PMID- 8408542 TI - Gene probes for identification of the botulinal neurotoxin gene and specific identification of neurotoxin types B, E, and F. AB - A polymerase chain reaction method was developed for the specific detection of the botulinum neurotoxin (BoNT) gene of Clostridium botulinum. Degenerate oligonucleotide primers, designed from the nucleotide sequence of the heavy chain of the BoNT gene, amplified a specific fragment of approximately 1.1 kb from strains of C. botulinum toxin types A, B, E, F, and G and neurotoxin-producing strains of Clostridium barati and Clostridium butyricum, but no fragment was obtained from nontoxigenic strains. The fragments amplified from several strains of C. botulinum types B, E, and F were cloned in Escherichia coli and their nucleotide sequences were determined. Sequences within this region were used to design oligonucleotide probes specific for BoNT type B (BoNT/B), BoNT/E, and BoNT/F genes. An additional probe was designed for the detection of the BoNT/F gene of C. barati, which differed in sequence from BoNT/F genes of both proteolytic and nonproteolytic strains of C. botulinum. PMID- 8408543 TI - Hybridization probes for conventional DNA fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of Cryptococcus neoformans. AB - In conventional DNA fingerprinting, hypervariable and repetitive sequences (minisatellite or microsatellite DNA) are detected with hybridization probes. As demonstrated here, these probes can be used as single primers in the polymerase chain reaction (PCR) to generate individual fingerprints. Several conventional DNA fingerprinting probes were used to prime the PCR, yielding distinctive, hypervariable multifragment profiles for different strains of Cryptococcus neoformans. PCR fingerprinting with the oligonucleotide primers (GTG)5, (GACA)4, and the phage M13 core sequence (GAGGGTGGXGGXTCT), but not with (CA)8 or (CT)8, generated DNA polymorphisms with all 42 strains of C. neoformans investigated. PCR fingerprints produced by priming with (GTG)5, (GACA)4, or the M13 core sequence differentiated the two varieties of C. neoformans, C. neoformans var. neoformans (serotypes A and D) and C. neoformans var. gattii (serotypes B and C). Furthermore, strains of serotypes A, D, and B or C could be distinguished from each other by specific PCR fingerprint patterns. These primers, which also successfully amplified hypervariable DNA segments from other species, provide a convenient method of identification at the species or individual level. Amplification of polymorphic DNA patterns by PCR with these primers offers several advantages over classical DNA fingerprinting techniques, appears to be more reliable than other PCR-based methods for detecting polymorphic DNA, such as analysis of random-amplified polymorphic DNA, and should be applicable to many other organisms. PMID- 8408544 TI - Phenotypical and genotypical characterization of epidemic clumping factor negative, oxacillin-resistant Staphylococcus aureus. AB - A total of 50 oxacillin-resistant Staphylococcus aureus (ORSA) strains that were clumping factor negative (CFN) and protein A negative by latex agglutination were collected from patients in six different hospitals at different locations in Germany during 1991 and 1992. Antibiograms, bacteriophage typing, and plasmid analysis were performed. The antibiograms showed that, besides oxacillin, all CFN ORSA strains were resistant to gentamicin, clindamycin, erythromycin, ciprofloxacin, and fosfomycin. All these isolates were nontypeable with an international set of phages, and an additional experimental phage set indicated that the strains were phage type 16, 192. Moreover, all isolates possessed a single plasmid of 30 kb, and restriction analysis of those plasmids revealed identical patterns. For genotyping, these 50 isolates were also analyzed by pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction (PCR) of the coagulase and protein A genes and then by restriction enzyme digestion and analysis of restriction fragment length polymorphisms (RFLPs). With 49 strains, electrophoresis of SmaI-digested chromosomal DNA revealed identical PFGE patterns regarding the number and size of the DNA fragments, which could be differentiated from those of clumping factor-positive ORSA strains. Typing for the coagulase gene by PCR revealed PCR products of identical sizes. The AluI restriction digestion patterns of the PCR products were identical. PCR with primers derived from the region of that part of the protein A gene that encodes the immunoglobulin G-binding domains showed a PCR product that was about 170 bp smaller than that of the protein A gene from strains that were positive in the protein A latex agglutination test. Since it is precisely this size that is required in order to encode one immunoglobulin G-binding region, we assume that this is not present in the CFN ORSA strains. The phenotypical and genotypical features identify these very unusual CFN ORSA stains as being of clonal origin. PMID- 8408545 TI - Comparison of rapid diagnostic techniques for respiratory syncytial and influenza A virus respiratory infections in young children. AB - We performed virus isolation tests for respiratory viruses on combined nasal wash throat swab specimens collected from infants and children with acute respiratory illnesses presenting to a hospital clinic during a 3-month period of concurrent epidemics of respiratory syncytial virus (RSV) and influenza A virus (Flu A) infections. Virus isolation results were used to assess the utility of commercially available rapid diagnostic kits for these two viruses. The kits employed direct immunofluorescence (IF) of cells (Imagen for RSV and Flu A), indirect IF of cells (Baxter Bartels Microscan), and enzyme immunoassay (EIA) (Becton Dickinson Directigen for RSV and Flu A and Abbott TestPack for RSV). All testing was completed on 81 specimens from 80 subjects. Of the 81 specimens, 53 (65%) yielded a virus: RSV, 28%; Flu A, 25%; rhinovirus, 6%; and enterovirus, cytomegalovirus, herpes simplex virus, and adenovirus, 2 to 4% each. Among the tests, Bartels Microscan and Directigen Flu-A exhibited the highest sensitivities (87 and 75%) and efficiencies (94 and 94%) for RSV and Flu A, respectively. All the tests exhibited high specificity. Thus, optimal detection of RSV and Flu A among infants and children who presented to a hospital clinic required two different detection methods (IF and enzyme immunoassay) and kits from two different companies (Baxter [Bartels Microscan] and Becton Dickinson [Directigen]). PMID- 8408546 TI - Combining pooling and alternative algorithms in seroprevalence studies. AB - Data from two seroprevalence studies and one comparative study of confirmatory algorithms were used to compare the costs and sensitivities of six algorithms for determining seropositivity to human immunodeficiency virus (HIV). We evaluated confirmatory strategies by using the CBC Recombigen HIV enzyme immunoassay (EIA; Cambridge BioScience, Worcester, Mass.) and immunoblotting followed by radioimmunoprecipitation assay to confirm indeterminate immunoblotting results with and without pooling of samples during screening. The least expensive algorithm was that in which sera were pooled during screening and EIA was used to confirm positive test results. The cost savings associated with this confirmatory test were greater when the prevalence of HIV infection was higher. Savings from pooling of sera for screen testing diminished as HIV prevalence increased. The sensitivity and specificity of EIA with respect to immunoblotting and radioimmunoprecipitation assay were estimated to be 0.9992 and 0.9977, respectively. We found that the implementation of pooling during screening and the use of EIA as the confirmatory test do not affect the statistical reliability of estimates of seropositivity but do result in considerable cost savings. PMID- 8408547 TI - Characterization of the pseudorabies virus-specific immunoglobulin M response and evaluation of its diagnostic use in pigs with preexisting immunity to the virus. AB - Despite preexisting immunity to pseudorabies virus (PRV), pigs may become infected and may or may not show clinical signs of disease. To investigate whether detection of immunoglobulin M (IgM) antibodies to PRV is suitable for diagnosis of recent infection in pigs with (or without) preexisting immunity, the IgM responses of pigs were examined after both experimental and natural infections. Upon inoculation of seronegative pigs with a low dose of a mildly virulent strain of PRV, IgM was first detectable at day 7 postinoculation (p.i.), reached a maximum at day 14 p.i., and became undetectable again at about days 32 to 36 p.i. In inoculated pigs with maternal antibodies against PRV, the IgM response began later and ended sooner, and peak titers were also lower. In immune pigs with maternally derived antibodies, there was apparently no correlation between the virulence of the inoculated strain and the IgM response. The suitability of the IgM enzyme-linked immunosorbent assay (ELISA) for detection of recent infection in the field was compared with that of the virus neutralization (VN) assay and with an ELISA which specifically detects antibodies directed to glycoprotein I (gI) of PRV. Paired sera were obtained from pigs suspected of PRV infection in an area endemic for PRV infection in which vaccination against PRV is often applied. Practically all pigs had antibodies to PRV in the acute phase of the disease. Compared with the VN assay, the specificity of the IgM ELISA was high but its sensitivity was low. However, all three serotests apparently failed to detect some PRV infections. The IgM ELISA appeared to be especially useful as a diagnostic aid for detection of recent infections in pigs with high levels of neutralizing and gI antibodies, probably maternally derived, in the acute phase of the disease. Such pigs may fail to develop a significant rise in VN antibody titer. The IgM ELISA may be the only serotest for monitoring infections in such pigs. PMID- 8408548 TI - Rapid competitive enzyme-linked immunosorbent assay using a monoclonal antibody reacting with a 15-kilodalton tegumental antigen of Schistosoma mansoni for serodiagnosis of schistosomiasis. AB - A competitive enzyme-linked immunosorbent assay (CELISA) for antibody detection was developed by using a monoclonal antibody which reacts with a 15-kDa tegumental antigen of the adult worm of Schistosoma mansoni. This monoclonal antibody was not able to react with antigens of Schistosoma japonicum or Schistosoma haematobium in enzyme-linked immunoelectrotransfer blot (EITB) and indirect immunofluorescence tests. The assay was performed in a period of 1 h using an adult worm crude extract antigen. To evaluate the CELISA, a total of 73 serum samples was analyzed: 35 were from S. mansoni-infected patients, 23 were from individuals with parasitic infections other than schistosomiasis, and 14 were from healthy individuals. All serum samples from healthy individuals and from patients infected with other parasites were negative, as were two (6%) samples from patients infected with S. mansoni. EITB analysis showed that 32 of 33 CELISA-positive samples were positive in the EITB but with different patterns of reactivity. A 15-kDa protein reacted with 60% of serum samples, and a 60-kDa protein showed the highest level of reactivity (85%). The two samples from patients infected with S. mansoni that were negative in the CELISA reacted with 70-, 60-, 50-, 47-, and 38-kDa proteins. One sample, positive in CELISA, did not react with proteins of the antigenic extract. PMID- 8408549 TI - Genome macrorestriction analysis of diversity and variability of Pseudomonas aeruginosa strains infecting cystic fibrosis patients. AB - Genome macrorestriction fingerprinting with XbaI and DraI was used to analyze the relatedness of 166 Pseudomonas aeruginosa isolates collected from 31 cystic fibrosis patients over a 1- to 20-month period and to correlate their genotype with patterns of resistance to 14 antimicrobial agents. Quantitative comparison of intra- and interpatient similarities of P. aeruginosa macrorestriction patterns disclosed two discrete ranges that clearly discriminated subclonal variation (> 80% relatedness) and clonal diversity (10 to 70% relatedness). Cloning-derived mutants exhibited up to 20% divergence of genomic macrorestriction patterns during the course of chronic colonization of individual patients. Change of susceptibility to multiple antimicrobial agents developed in 50% of sequential pairs of isolates from individual patients. Only 19% of these susceptibility changes were attributable to strain substitution, while the majority (56%) of resistance changes were associated with minor genomic variations of a persistent strain. Sixty-six percent of patients harbored one strain, and 33% carried two strains. Three common strains colonized 5 (28%) of 18 patients attending a cystic fibrosis clinic, and another two strains colonized two patient pairs (31%) of 13 patients staying at a rehabilitation center, suggesting potential cross-infection in these settings. By indexing regional polymorphisms throughout the chromosome structure, macrorestriction analysis can monitor subclonal evolution of P. aeruginosa and identify isogenic resistance mutants. Quantitative macrorestriction fingerprinting enables discrimination between clonal variants and clones of distinct origins and should therefore provide a reliable tool for investigating the mode of acquisition of P. aeruginosa in cystic fibrosis patients. PMID- 8408550 TI - Diagnosis of Toxoplasma parasitemia in patients with AIDS by gene detection after amplification with polymerase chain reaction. AB - We sought evidence of toxoplasma parasitemia among 37 people with active or dormant Toxoplasma gondii infection or no evidence of infection. DNA was extracted from erythrocyte-free portions of blood samples, and the T. gondii B1 gene was amplified by the polymerase chain reaction. Evidence of T. gondii parasitemia was found in six patients with severe immunosuppression from AIDS and clinical evidence suggestive of or compatible with toxoplasmosis. Results were negative for patients unlikely to have active toxoplasmosis. Gene detection after amplification with the polymerase chain reaction is a promising test for detection of parasitemia, and parasitemia should be tested for in patients with AIDS and unexplained fever or central nervous system abnormalities. PMID- 8408551 TI - Rapid broth macrodilution method for determination of MICs for Mycobacterium avium isolates. AB - A multicenter study was done to investigate the accuracy and reproducibility of a method for determining the MICs of antimicrobial agents against the Mycobacterium avium complex in 7H12 broth with the BACTEC system. In phase I, with eight drugs and 10 strains, intralaboratory reproducibility was 95.7 to 100%, allowing a 1 dilution difference upon repeat testing. The results of phase II testing with 41 additional strains were consistent with those obtained in phase I, with good interlaboratory reproducibility. The radiometric method was validated by sampling and plating of the same broth cultures and determining, by the number of CFU per milliliter, the lowest drug concentration that inhibited more than 99% of the initial bacterial population. Three test concentrations of each drug and the tentative interpretation of results are proposed. Radiometric MIC determination has the potential to become the method of choice for clinical microbiology laboratories and evaluation of new agents for the treatment of M. avium infections, both pulmonary and disseminated. PMID- 8408552 TI - Evaluation of counterimmunoelectrophoresis for serotyping Actinobacillus pleuropneumoniae isolates and detection of type-specific antigens in lungs of infected pigs. AB - A rapid, simple, and accurate counterimmunoelectrophoresis technique was developed for serotyping cultures of Actinobacillus pleuropneumoniae as well as for detection of their type-specific antigens in the lung tissues of infected pigs. The counterimmunoelectrophoresis test correctly identified all of the reference antigens and more than 99% of 1,200 field isolates of A. pleuropneumoniae representing the 12 established serotypes within 1 h. Counterimmunoelectrophoresis and coagglutination tests did not differ broadly in sensitivity from each other. Both procedures were more rapid and more sensitive than immunodiffusion and indirect hemagglutination tests. A total of 355 lung tissue samples (130 lungs of pigs that died because of acute respiratory problems, 125 lungs of pigs from herds with chronically infected pleuropneumonia, and 100 lungs from apparently healthy pigs at the slaughterhouse) were examined for the presence of A. pleuropneumoniae type-specific antigens by counterimmunoelectrophoresis, coagglutination, and immunodiffusion tests. A. pleuropneumoniae type-specific antigen was found in all 55 samples from which the bacteria had earlier been isolated and in 27 specimens in which they had not been found. Detection of antigen in the lung tissues by coagglutination and counterimmunoelectrophoresis tests was found to be much simpler and much more rapid than conventional culture isolation. Both counterimmunoelectrophoresis and coagglutination tests were found extremely useful in the diagnosis of acute cases of porcine pleuropneumonia. However, these techniques were able to detect only some of the chronically infected carrier pigs. PMID- 8408553 TI - Immune response to synthetic peptides of hepatitis delta antigen. AB - Hepatitis delta antigen (HDAg) is the only viral protein known to be expressed during hepatitis delta virus (HDV) infection. Detection of antibody to HDAg (anti HD) is the usual method for diagnosis of HDV infection since viremia lasts only a few weeks. In an effort to identify the major epitopes recognized by humans during natural infection, four oligopeptides including residues 2 to 17 (SP1), 155 to 172 (SP2), 168 to 182 (SP3), and 189 to 211 (SP4) of the HDAg molecule were synthesized and probed by enzyme-linked immunosorbent assay with a panel of 80 serum specimens from 45 patients suffering from either HDV-hepatitis B virus coinfections (n = 17) or HDV superinfections (n = 28). Sera from infected patients recognized these relatively short peptides. Peptide SP2 was the most antigenic; 71% of serum specimens reacted. Antibody to SP2 was also the commonest in sera taken early in the course of the disease. Peptide SP2 corresponds to one of the two regions which is highly conserved between different isolates. Among the 63 serum specimens which scored anti-HD positive by a commercial assay, all but 3 reacted to at least one of the peptides (95% agreement). Peptide assays appeared to be significantly more sensitive than the commercial assay with native HDAg early in the course of HDV infection since 14 of 17 (82%) serum specimens which scored anti-HD negative in the commercial assay reacted to one or more peptides. All serum specimens giving one or more positive results with the various peptides were confirmed as being HDV positive by an inhibition assay with free peptide in solution. The immune response to HDAg peptides vared greatly between individuals. No specific reactivity profile could be assigned to those with either HDV-hepatitis B virus coinfections or HDV superinfections. Overall, HDAg peptides appeared to be convenient reagents in addition to native antigen for the development of new and improved diagnostic tests for HDV infection. PMID- 8408554 TI - Expression of feline immunodeficiency virus gag and env precursor proteins in Spodoptera frugiperda cells and their use in immunodiagnosis. AB - The gag and env genes of the feline immunodeficiency virus strain UT113 were cloned into a baculovirus transfer vector. The recombinant plasmids were used to create recombinant baculoviruses that expressed either the gag or the env precursor protein in insect cells (Sf9 cells). Leader sequence cleavage occurred in Sf9 cells expressing the envelope precursor, but further processing was not observed. Crude lysates of insect cells infected with the wild-type baculovirus or with the recombinant viruses were used to develop an enzyme-linked immunosorbent assay for the detection of feline immunodeficiency virus-specific antibodies in cat sera. The assay showed a higher sensitivity and specificity than immunofluorescence and Western blotting (immunoblotting). PMID- 8408555 TI - Preventing false positives: quantitative evaluation of three protocols for inactivation of polymerase chain reaction amplification products. AB - False-positive results because of carryover contamination by previously amplified nucleic acids are currently the greatest impediment to routine implementation of nucleic acid amplification protocols. We evaluated three methods for inactivation of a 156-bp Borrelia burgdorferi polymerase chain reaction (PCR) product: (i) post-PCR cross-linking with isopsoralen (IP), (ii) pre-PCR treatment of a dU containing PCR product with uracil N-glycosylase (UNG), and (iii) post-PCR alkaline hydrolysis (primer hydrolysis) of PCR products synthesized by using primers containing 3' ribose residues. The sensitivities of the PCR performed under the conditions of each protocol were comparable. Inactivation of amplified DNA was highly efficient for all three protocols; the IP and UNG protocols eliminated at least to 3 x 10(9) copies of the product. The primer hydrolysis protocol varied in efficiency depending on the number and position of the 3' ribose residues, but inactivation ranged from 10(4) to 10(9) copies. We conclude that with some modifications, all three systems are effective for eliminating amplified DNA products. Routine implementation of at least one method should help to avoid false-positive results because of carryover contamination. PMID- 8408556 TI - Dependence of polymerase chain reaction product inactivation protocols on amplicon length and sequence composition. AB - Specific diagnostic test results generated by polymerase chain reaction (PCR) depend upon control of amplicon contamination in the clinical laboratory. We compared photochemical (isopsoralen [IP]) and enzymatic (uracil N-glycosylase [UNG]) methods for their ability to prevent carryover of amplicons generated from genomic targets of five viruses. PCR products (amplicons) (herpes simplex virus, 342 bp; cytomegalovirus, 250 bp; Epstein-Barr virus, 240 bp) exposed to UV light in the presence of various concentrations of IP compound 10 (IP-10) resulted in apparent increased molecular sizes of the products, as indicated by migration patterns after gel electrophoresis, and were predictive of inactivation by the agent. For amplicons of < or = 100 bp, IP-10-induced electrophoretic shifts were related to the guanidine-cytidine (G + C) content of the PCR product; no apparent shift and no inactivation were observed for a 92-bp herpes simplex virus amplicon (G + C content, 65%), whereas the 100-bp human papillomavirus product (G + C content, 42%) showed a concentration-dependent shift (25 to 100 micrograms/ml) in electrophoretic migration and was partially inactivated. UNG effectively controlled amplicon carryover for target DNA of > or = 240 bp; however, this treatment did not inactivate the two amplicons of < or = 100 bp, regardless of the G + C content of the product. Larger products were inactivated efficiently by both methods, regardless of their G + C contents. We concluded that both IP and UNG effectively inactivated PCR amplicons but not short amplicons of < or = 100 bp. We recommend that with the adoption of PCR technology in clinical laboratories, primers should be designed to produce amplicons of at least 240 to 350 bp (depending on G + C content) and that at least one effective method of controlling carryover contamination should be incorporated into each PCR protocol. PMID- 8408557 TI - Influence of zinc on Pseudomonas aeruginosa susceptibilities to imipenem. AB - Serial dilution susceptibility testing of imipenem against 59 clinical isolates of Pseudomonas aeruginosa, conducted simultaneously on single lots of Difco and BBL Mueller-Hinton agar (MHA), resulted in MICs for 90% of strains tested of 8 and 16 micrograms/ml, respectively. MICs for Escherichia coli, Klebsiella pneumoniae, and Pseudomonas spp. were also higher on BBL MHA. Quantification of the cation content of the two MHAs by atomic absorption spectroscopy demonstrated that the zinc concentration in BBL MHA was 15 times greater than that measured in Difco MHA (2.61 and 0.17 micrograms/ml, respectively). Concentrations of calcium, magnesium, iron, manganese, and copper in the two agars were similar. Addition of zinc to Difco MHA resulted in increases in MICs of imipenem for P. aeruginosa but not in the MICs of ceftazidime or cefpirome for P. aeruginosa (P < 0.01). A lesser zinc effect was seen on the activity of imipenem against E. coli, K. pneumoniae, and Pseudomonas spp. The activities of ceftazidime and cefpirome were similar on both MHAs when tested against all gram-negative organisms in this study. Thus, the effect of zinc in MHA was clearly demonstrated by a significant increase in the MICs of imipenem for P. aeruginosa, and, to a lesser extent, for other gram-negative bacilli. PMID- 8408558 TI - Evaluation of sputum smears concentrated by cytocentrifugation for detection of acid-fast bacilli. AB - Early identification and isolation of tuberculosis patients is of utmost importance to minimize the risk of further epidemic spread of the disease. The traditional concentrated acid-fast smears are not very reliable tools for the presumptive diagnosis of tuberculosis. Acid-fast bacillus (AFB) smears from 120 patients specimens and 80 simulated AFB samples were processed according to standard laboratory procedures and by cytocentrifugation (Cyto-Tek, Ames Division, Miles Laboratories, Inc., Elkhart, Ind.). Prior to dispensing of samples into the Cyto-Tek chambers, specimens were liquefied and decontaminated by mixture with an equal volume of 5% sodium hypochlorite (household bleach). Culture and smear results were correlated. Of 120 patient specimens, 43 were culture and smear negative by both methods. Ten specimens were overgrown with mold and bacteria, but 2 of them had positive AFB smears by cytocentrifugation only. There were 67 positive AFB cultures, with 67 positive cytocentrifuge smears and 34 positive smears by the conventional technique. Of the 80 simulated positive AFB samples, all grew mycobacteria on culture. Smears from the 10(5)- to 10(3)-CFU/ml specimens were positive by both methods. The simulated samples with 10(2) CFU/ml yielded smears positive only by cytocentrifugation. The Cyto-Tek AFB smears had a greater correlation with positive culture than did the smears from concentrated specimens. The sensitivity, efficiency, and rapidly of the Cyto-Tek AFB smear technique resulted in increased detection of mycobacteria in clinical specimens. The simplicity and safety of this method will enable qualified mycobacteriology technologists to rapidly and accurately perform sputum smears for AFB at clinics, emergency rooms, and field sites, as well as in the traditional laboratory setting. PMID- 8408559 TI - Tentative criteria for confirming the in vitro susceptibilities of Haemophilus influenzae and Neisseria gonorrhoeae to two fluoroquinolones (sparfloxacin and levofloxacin), including quality control parameters. AB - Sparfloxacin and levofloxacin were evaluated against 150 Haemophilus influenzae isolates and 149 Neisseria gonorrhoeae isolates in order to define susceptibility testing parameters. Sparfloxacin-susceptible H. influenzae strains were defined as those for which the MICs were < or = 0.25 microgram/ml and the zones were > or = 30 mm, and N. gonorrhoeae susceptible strains were those for which the MICs were < or = 0.03 microgram/ml and the zones were > or = 39 mm (5-micrograms disks). Levofloxacin-susceptible strains of H. influenzae included those for which the MICs were < or = 0.12 microgram/ml and the zones were > or = 32 mm and N. gonorrhoeae susceptible strains were those for which the MICs were < or = 0.12 microgram/ml and the zones were > or = 37 mm (5-micrograms disks). Criteria for a resistant category cannot yet be defined for either quinolone. In multilaboratory studies with different lots of Haemophilus Test Medium, replicate tests with the standard control strain of H. influenzae (ATCC 49247) were evaluated. For sparfloxacin disk tests, the proposed zone size limits were 33 to 42 mm and broth microdilution MIC limits were 0.004 to 0.016 microgram/ml, whereas for levofloxacin tests, zone size limits were 32 to 41 mm and broth microdilution MIC limits were 0.008 to 0.03 microgram/ml. Other multilaboratory studies evaluated tests with supplemented GC agar and N. gonorrhoeae ATCC 49226; for both drugs, zone size limits were 44 to 52 mm and agar dilution MIC limits were 0.004 to 0.016 microgram/ml. PMID- 8408560 TI - Bacteremia due to Rochalimaea henselae in a child: practical identification of isolates in the clinical laboratory. AB - Two closely related species of Rochalimaea, Rochalimaea quintana and Rochalimaea henselae, are nutritionally fastidious but can be cultivated on bacteriologic media from the blood of patients with diverse clinical presentations. We report a case of culture-proven R. henselae bacteremia in a child with persistent fever. Serologic evidence of infection by R. henselae was ascertained by testing sera at two intervals for immunoglobulin G or immunoglobulin M antibodies by enzyme immunoassay and immunoblot. The case isolate and a collection of other strains (R. henselae, R. quintana, and related organisms) were used to test commercial identification systems for their comparative utility in the identification of Rochalimaea spp. on a practical basis. Of six systems designed for testing of either fastidious or anaerobic isolates of bacteria, the MicroScan Rapid Anaerobe Panel was the only system that distinguished R. henselae from R. quintana. Four of five others gave reactions that were unique within their data bases but did not distinguish Rochalimaea isolates at the species level. PMID- 8408561 TI - Paecilomyces variotii in peritoneal dialysate. AB - Four cases of peritonitis caused by the filamentous fungus Paecilomyces variotii in patients on continuous ambulatory peritoneal dialysis are reported. Removal of the Tenckhoff catheter and antifungal chemotherapy led to resolution of symptoms in all cases. Possible contaminating events are discussed, and reported infections with P. variotii are reviewed. PMID- 8408562 TI - Automated interpretation of disk diffusion antibiotic susceptibility tests with the radial profile analysis algorithm. AB - An original algorithm referred to as the radial profile analysis algorithm was implemented on a Macintosh Quadra 700 computer to provide an automatic determination of the inhibition zone diameters of antibiotic susceptibility tests performed with the disk diffusion method. After digitization of the petri plate image, each antibiotic disk is recognized and labeled. Pixels of the local zone around each disk are then used for generating a profile pattern that is subjected to decision rules. The resulting estimate of the inhibition zone diameter is then automatically compared with conventional breakpoints for classifying the tested strain in one of the clinical categories of antibiotic susceptibility. The program is also able to request a human reading for some rare plates difficult to interpret. The algorithm accuracy was tested by comparing the results with a combination of independent human measurements performed on the tested plates. The test sample was composed of 98 strains, and 2,552 tests of 40 distinct antibiotics were subjected to the analysis. The difference between the automatic and human diameter estimates was less than 4 mm in 90% of the tests. The agreement between the automatic and human clinical categorizations amounted to 95.5%, and severe (major and very major) disagreements were found in 5.6% of the tests performed with staphylococci but only 0.3% of the tests with gram-negative rods. We conclude that the radial profile analysis algorithm is a solid backbone for an automatic system dedicated to the clinical interpretation of disk diffusion antibiotic susceptibility tests. PMID- 8408563 TI - Sensitive enzyme-linked immunosorbent assay for detection of Clostridium botulinum neurotoxins A, B, and E using signal amplification via enzyme-linked coagulation assay. AB - A new immunoassay amplification method has been applied to the measurement of toxins A, B, and E from Clostridium botulinum. The technique is a modified enzyme linked immunosorbent assay (ELISA) which relies on the detection of sandwich complexes on microtiter plates by a solid-phase coagulation assay known as ELCA, or enzyme-linked coagulation assay. In the method, a coagulation activating enzyme (RVV-XA) isolated from the venom of Russell's viper is conjugated to affinity-purified horse antibodies specific for toxin type A, B, or E. Plates are coated with affinity-purified antibodies, and standard captag (capture-tag) protocols using labeled antibody are employed to bind the toxin from solution. Complexes are detected by adding a modified plasma substrate which contains all the coagulation factors mixed with alkaline phosphatase-labeled fibrinogen and solid-phase fibrinogen; deposition of solid-phase, enzyme-labeled fibrin on the solid phase is then a reflection of formation of toxin-RVV-XA-antibody complexes on the solid phase. Because of the ability to detect RVV-XA by this coagulation assay at concentrations < 0.1 pg/ml, it was possible to measure C. botulinum toxins A, B, and E at mouse bioassay levels (< 10 pg/ml, or < 0.07 pM) for both purified neurotoxin and crude culture filtrates obtained from strains known to produce appropriate single toxins. ELISA-ELCA should be applicable to measurement of toxins in most of the materials (contaminated food, blood, and excreta) for which the comparably sensitive mouse bioassay is currently employed. This method has the potential of broad application to the measurement of low concentrations of any antigen for which appropriate immunochemical reagents are available, in a color test format. PMID- 8408564 TI - Detection and identification of Mycobacterium tuberculosis directly from sputum sediments by amplification of rRNA. AB - Seven hundred fifty-eight processed sputum sediments received for the diagnosis of tuberculosis or other mycobacterial infections were tested by utilizing a rRNA target amplification assay and traditional culture techniques. The results from the rRNA target amplification assay (Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test), available in 5 h, were compared with the results from standard culture techniques held for 6 weeks. A total of 119 specimens (16%) were culture positive for Mycobacterium tuberculosis. Overall sensitivity, specificity, positive predictive value, and negative predictive value were 82, 99, 97, and 96%, respectively, for the Gen-Probe assay; 88, 100, 100, and 97%, respectively, for culture; and 53, 99.8, 99.6, and 91%, respectively, for fluorochrome stain. The Gen-Probe assay employs the isothermal enzymatic amplification of M. tuberculosis complex rRNA followed by detection of the amplicon with an acridinium ester-labeled DNA probe. This assay has the potential of reducing the time for diagnosis of tuberculosis to 1 day. PMID- 8408565 TI - Epidemiological study of an Acinetobacter baumannii outbreak by using polymerase chain reaction fingerprinting. AB - A polymerase chain reaction (PCR) technique was applied to the fingerprinting of different strains of Acinetobacter baumannii from a cluster of patients infected or colonized with the incriminated pathogen. The DNA was extracted by boiling and was subjected to PCR amplification by using the core sequence of the M13 phase as a single primer. The amplified products were separated by agarose gel electrophoresis and were detected by staining with ethidium bromide. In 1990, 49 multiresistant A. baumannii strains were isolated from 13 patients from the same intensive care unit of the Charite Hospital; 45 of these outbreak isolates obtained from 12 patients showed the same PCR patterns, indicating an epidemiological relatedness of these strains. Four strains isolated from the same patient belonged to another genetic group, as revealed by a distinct amplification pattern. Another single subtype of A. baumannii was identified as the causative agent in patients during a second outbreak at a different intensive care unit in the same hospital. Seventeen isolates recovered from 10 immunocompromised patients had the same amplification patterns, which were distinct from all other PCR profiles. Five strains were obtained from two other hospitals; three isolates from the hospital of Magdeburg, Germany, had identical PCR patterns which, however, could be clearly distinguished from the patterns of all other strains. The remaining two isolates displayed individual patterns of amplified fragments. PCR fingerprinting may provide a useful and particularly rapid identification technique for epidemiological investigations of nosocomial infections. PMID- 8408566 TI - Detection of Trypanosoma cruzi in blood specimens of chronic chagasic patients by polymerase chain reaction amplification of kinetoplast minicircle DNA: comparison with serology and xenodiagnosis. AB - A panel of 114 blood samples from chronic chagasic patients and nonchagasic patients was screened for Trypanosoma cruzi by xenodiagnostic, serologic, and polymerase chain reaction (PCR) amplification tests. Blood samples were preserved in a guanidine-EDTA buffer, and total blood DNA was isolated after chemical nuclease cleavage with 1,10-phenanthroline-copper ion and used as a template for PCR amplification of the conserved and variable regions of T. cruzi minicircle molecules. The PCR products were screened by Southern blot hybridization with a digoxigenin-labeled oligonucleotide probe specific for the conserved region of the minicircle. The method showed a sensitivity of 100% compared with the serologic test. In addition, all of the serology-positive, xenodiagnosis-negative samples were positive by PCR. This demonstrates that PCR amplification of T. cruzi kinetoplast minicircle DNA could replace xenodiagnosis for evaluation of parasitemia in chronic chagasic patients and could serve as a complement for serologic testing in the screening of blood bank donors. PMID- 8408567 TI - Intrathecal immune response and virus-specific immunoglobulin M antibodies in laboratory diagnosis of acute poliomyelitis. AB - The intrathecal immune response in 114 patients with clinically diagnosed acute poliomyelitis was studied by measuring poliovirus-specific immunoglobulin M (IgM) antibodies in cerebrospinal fluid (CSF) by a mu-capture immunoassay and by assessing the ratio between levels of poliovirus-neutralizing antibodies in serum and CSF. Fecal specimens were used for attempts to isolate the causative agents. Eighty-five percent of CSF specimens collected during the first 15 days of disease contained virus-specific IgM antibodies. Forty-five of 48 tested children (94%) also showed virus-specific IgM responses in their sera. Later on, the antibody levels decreased, and positive results after 30 days of onset of paralytic symptoms were rare. If the presence of poliovirus-specific IgM antibodies in the CSF was considered diagnostic, more cases were confirmed by this test than by virus isolation. A relative increase in poliovirus-neutralizing antibodies in the CSF was observed in about one-third of the cases; in all but three cases the increase was observed together with the presence of virus specific IgM antibodies. A systemic virus-specific response can be seen and poliovirus can be isolated from a subclinically infected individual suffering from a concomitant poliomyelitis-like disease, while positive results by the two methods demonstrating an intrathecal immune response are likely to indicate a true causal relationship between infection and disease. Demonstration of poliovirus-specific IgM antibodies in the CSF thus appears to be a sensitive and specific method for laboratory confirmation of clinically diagnosed poliomyelitis. PMID- 8408568 TI - Analysis of acquired human cytomegalovirus infections by polymerase chain reaction. AB - We used the polymerase chain reaction and primers corresponding to three regions of the human cytomegalovirus (HCMV) genome to study HCMVs isolated from 16 children attending a single day-care center and the father of two children in the same center. When we analyzed isolates with primers for the pp65 and major immediate-early genes, we observed nearly uniform amplification yielding products of predicted sizes. By contrast, primers for the a sequence demonstrated variability among HCMV strains, supporting the use of these primers as an epidemiologic tool. Analysis of a-sequence products from two isolates demonstrated 50 to 70% nucleotide homology with the a sequence of HCMV Towne strain DNA. We observed 95% nucleotide homology for the two a-sequence products derived from the father-child pair. Analysis of day-care center isolates indicated that two children excreted two distinct HCMV strains during the study interval. PMID- 8408569 TI - Reactogenicity and immunogenicity of a high-titer rhesus rotavirus-based quadrivalent rotavirus vaccine. AB - We evaluated the reactogenicity and antigenicity of a quadrivalent rotavirus vaccine composed of serotype 3 rhesus rotavirus (RRV) and three single-gene substitution reassortants of RRV and human strain D (D x RRV, serotype 1), DS1 (DS1 x RRV, serotype 2), or ST3 (ST3 x RRV, serotype 4) in a double-masked study with 302 infants in Caracas, Venezuela. Three doses of the quadrivalent vaccine composed of either 10(5) PFU (low titer) or 10(6) PFU (high titer) of each component were administered to 99 and 101 infants, respectively, at 4-week intervals starting at the second month of age; 102 infants received a placebo. Postvaccination reactions were monitored by home visits every other day during the week postvaccination. The vaccine was associated with the occurrence of mild, short-lived febrile episodes in 26 and 23% of the recipients after the first doses of high- or low-titer vaccine, respectively, in comparison with 13% of the infants receiving the placebo. Febrile reactions occurred less frequently in vaccinees after the second or third dose than after the initial dose. The vaccine was not significantly associated with diarrhea or any additional symptom or sign. Serum specimens obtained shortly before the first, 4 weeks after the first, and 4 weeks after the third dose of vaccine or placebo were tested by an immunoglobulin A enzyme-linked immunosorbent assay and by neutralization assays. Seroresponses occurred significantly more often after 3 doses than after a single dose of either vaccine. Immunoglobulin A responses were observed in 80 and 79% of the infants after 3 doses of high- or low-titer vaccine, respectively. Most of the infants tested developed a neutralization response to RRV after 3 doses of the high- (90%) or low-(88%) titer vaccine. Neutralization response rates to human rotavirus serotypes 1 to 4 after 3 doses were similar in both vaccine and 87 of 90 receiving the high-titer vaccine developed seroresponses, as detected by any of the assays employed. The study indicates that 3 doses of quadrivalent vaccine at a titer of 10(6) PFU of each component offered no advantage over the lower titer preparation for use in efficacy trials. PMID- 8408570 TI - Neisseria weaveri sp. nov., formerly CDC group M-5, a gram-negative bacterium associated with dog bite wounds. AB - CDC group M-5 is a rod-shaped, gram-negative, nonmotile bacterium associated with dog bite wounds. DNA-DNA relatedness and biochemical and growth characteristics were studied for 54 strains from the collection at the Centers for Disease Control and Prevention. One typical M-5 strain, 8142, was further studied by 16S rRNA sequencing. DNA from 40 of 53 strains showed 82 to 100% relatedness (hydroxyapatite method) to labeled DNA from strain 8142. The guanine-plus cytosine (G + C) content in 8 of the 41 highly related M-5 strains was 50.5 to 52 mol%. These 41 strains were oxidase and catalase positive, nonfermentative, nitrite positive, nitrate negative, weakly phenylalanine deaminase positive, aerobic, and alpha-hemolytic (sheep blood). DNA from the 13 remaining strains showed only 7 to 46% DNA relatedness to strain 8142. These 13 non-M-5 strains differed from the M-5 strains in G + C content, growth characteristics, and biochemical profiles. DNA from M-5 strain 8142 was most closely related to DNA from groups EF-4b (47%) and EF-4a (45%). 16S rRNA sequence analysis placed M-5 strain 8142 in the Neisseriaceae cluster of the beta-3 subgroup of the class Proteobacteria. It was most homologous (98.4 to 98.8%) to Neisseria animalis, Neisseria flavescens, Neisseria canis, and Neisseria elongata. All data are consistent with M-5 being a new species of Neisseria, for which we propose the name Neisseria weaveri. PMID- 8408571 TI - Prevalence and some properties of verotoxin (Shiga-like toxin)-producing Escherichia coli in seven different species of healthy domestic animals. AB - Fecal samples from 720 healthy, domestic animals representing seven different species (cattle, sheep, goats, pigs, chickens, dogs, and cats) were investigated for verotoxin (VT [Shiga-like toxin])-producing Escherichia coli (VTEC). VTEC were isolated from 208 animals (28.9%), most frequently from sheep (66.6% VTEC carriers), goats (56.1%), and cattle (21.1%). VTEC were isolated less frequently from pigs (7.5%), cats (13.8%), and dogs (4.8%) and were not found in chickens (< 0.7%). Forty-one different O:H serotypes and 23 untypeable O-groups were isolated. Five serotypes (O5:H-, O91:H-, O146:H21, O87:H16, and O82:H8) occurred in more than one animal species. Serotypes O5:H-, O91:H-, O146:H21, O128:H2, and OX3:H8 represented 54.8% of the VTEC strains. Nearly 60% of all VTEC O:H serotypes isolated in this study have been implicated as human pathogens, indicating that healthy, domestic animals may serve as a reservoir of human pathogens. All VTEC, except nine feline strains, hybridized with one or both of the VT1 and VT2 specific DNA probes. VT production and enterohemolysin (E-Hly+) production were associated in E. coli from goats, sheep, and cattle but not in E. coli from chickens, pigs, dogs, and cats. A close association of VT with E-Hly+ was found in O5:H-, O146:H21, O128:H2, O77:H4, O119:H25, and O123:(H10) strains. Thirty of 240 (12.5%) E-Hly+ strains hybridized with an E-Hly+ specific DNA probe, indicating heterogeneity of regulatory or structural E-Hly+ genes in strains of E. coli. PMID- 8408572 TI - New cause for false-positive results with the Pastorex Aspergillus antigen latex agglutination test. AB - The Pastorex Aspergillus antigen test for detection of Aspergillus galactomannan antigen in the sera of patients with invasive aspergillosis is used in many clinical laboratories. A serum sample contaminated with Penicillium chrysogenum gave a strongly positive reaction (1:128) which was heat stable, was not eliminated by pronase treatment, and was not detected by a normal rabbit globulin control. This observation was shown to be due to cross-reactions of the monoclonal antibody EB-A2 used by the kit with several airborne fungi likely to contaminate serum samples, including Penicillium chrysogenum, Cladosporium herbarum, Acremonium species, Alternaria alternata, Fusarium oxysporum, Wangiella dermatitidis, and Rhodotorula rubra. PMID- 8408573 TI - Detection of rotavirus serotypes G1, G2, G3, and G11 in feces of diarrheic calves by using polymerase chain reaction-derived cDNA probes. AB - On the basis of antigenic variability in the VP7 outer capsid glycoprotein, at least 14 G serotypes exist for group A rotaviruses. Serotypic diversity exists among bovine rotaviruses (BRV), with serotypes G1, G6, G8, and G10 reported for cattle. Although G1 and G8 rotaviruses were originally described for humans, the recent isolation of G6 and G10 rotaviruses from humans further emphasizes the serotypic similarity between human and bovine rotaviruses and the possible zoonotic potential of rotaviruses. Results of our previous studies have indicated that more than 24% of BRV-positive field samples from diarrheic calves were nonreactive with cDNA probes or monoclonal antibodies to serotypes G6, G8, and G10. In this study, cDNA probes were prepared by polymerase chain reaction amplification of the hyperdivergent regions of the VP7 genes (nucleotides 51 to 392) from human (G1, G2, and G3) and porcine (G4, G5, and G11) rotaviruses. These probes were used in a dot blot hybridization assay to further characterize the G types of 59 BRV strains (fecal samples from diarrheic calves in Ohio, Nebraska, Washington, and South Dakota) that were nonreactive with cDNA probes to G6, G8, and G10. Rotaviruses belonging to serotypes G1 (n = 7), G2 (n = 1), G3 (n = 2), and G11 (n = 3) were identified among the BRV field samples. The BRV associated with these G types accounted for 22% of the samples tested; the other 78% of these samples remained untypeable with these probes. To our knowledge, this is the first report in the United States of the identification among BRV isolates of rotavirus serotypes G1, G2, G3, and G11. PMID- 8408574 TI - Wound isolate of Salmonella typhimurium that became chlorate resistant after exposure to Dakin's solution: concomitant loss of hydrogen sulfide production, gas production, and nitrate reduction. AB - A strain of Salmonella typhimurium isolated from a decubitus ulcer that was being treated topically with half-strength Dakin's solution became H2S negative, nitrate negative, and unable to produce gas from glucose. Experimental data suggested that these effects were associated with the development of chlorate resistance. Thirty-five other strains of Salmonella spp. that were made chlorate resistant also became negative for these three tests. PMID- 8408575 TI - Identification of vancomycin-resistant lactic bacteria isolated from humans. AB - By using cell morphology, arginine dihydrolase, and gas production in de Man, Sharp, Rogosa broth, 122 isolates of vancomycin-resistant lactic bacteria from humans were assigned to five profiles, allowing us to distinguish Pediococcus, homofermentative and heterofermentative Lactobacillus, and Leuconostoc species. The absence of L-(+)-lactic acid, as detected spectrophotometrically, was confirmatory for Leuconostoc species. API 50 CHL panels were useful for the identification of Lactobacillus species. PMID- 8408576 TI - Typing of Aspergillus species and Aspergillus fumigatus isolates by interrepeat polymerase chain reaction. AB - Polymerase chain reaction amplification of repetitive DNA motifs allows discrimination of Aspergillus species. In a preliminary survey, the DNA fingerprints appear to be identical when isolates of Aspergillus fumigatus are compared. When a primer deduced from a prokaryotic repeat motif is used, Aspergillus fumigatus isolates originating from different patients or different anatomical locations can be typed individually. PMID- 8408577 TI - Genomic fingerprinting of Neisseria meningitidis associated with group C meningococcal disease in Canada. AB - A single electrophoretic type (ET15) of Neisseria meningitidis has been associated with an increased incidence of group C meningococcal disease in Canada. Genomic fingerprinting through pulsed-field gel electrophoresis of chromosomal DNA was used to characterize the clonal relationship among meningococcal isolates of different electrophoretic types and among isolates within ET15. The genomic fingerprints of the ET15 isolates, while similar as a group, were sufficiently distinct to confirm linkage for four pairs of strains from focal outbreaks and differed markedly from those of the other common electrophoretic types, ET5, ET9, and ET21. PMID- 8408578 TI - Cytopathic effect mimicking virus culture due to Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis grew, from a bronchoalveolar lavage of a patient with AIDS, on a gentamicin-supplemented cell culture monolayer, causing a focal and slowly spreading cytopathic effect resembling that of a virological isolate. The same effect was observed after inoculation of two different inocula of M. tuberculosis onto the same cell culture. PMID- 8408579 TI - False-negative cerebrospinal fluid cryptococcal latex agglutination tests for patients with culture-positive cryptococcal meningitis. AB - Three cases of false-negative cerebrospinal fluid latex agglutination test results for patients with culture-positive cryptococcal meningitis are reported. False-negative results occurred in settings of low cryptococcal antigen concentrations in cerebrospinal fluid and were dependent on the latex agglutination test kit used. Investigation of each case revealed that prozone phenomena or interference from bound antibody or protein could not account for the false-negative results. PMID- 8408580 TI - Mucor ramosissimus Samutsevitsch isolated from a thigh lesion. AB - Mucor ramosissimus Samutsevitsch is presented for the first time as an etiologic agent of cutaneous zygomycosis in a patient with aplastic anemia on immunosuppressive therapy. This report also represents the third case caused by this species reported in the literature. A biopsy taken from a lesion on the patient's thigh revealed broad, nonseptate, nonbranching hyphae compatible in morphology with a Zygomycete; M. ramosissimus was cultured twice from the thigh lesion. The patient was treated successfully with amphotericin B. Identifying features of M. ramosissimus include the following: numerous sporangia lacking columellae and resembling those of Mortierella spp., short, erect sporangiophores repeatedly branching sympodially; tough, persistent, and diffluent sporangial walls; numerous oidia in chains; extremely low colonies; and restricted growth at 36 degrees C. This paper describes the isolate and strives to alert the clinical microbiologist to this rarely reported pathogen. PMID- 8408581 TI - Use of an enzyme-linked immunoassay for Clostridium difficile serogrouping. AB - An enzyme-linked immunoassay (ELISA) with 11 Clostridium difficile serogroup specific antisera was applied for serogrouping of C. difficile colonies from 314 consecutive positive fecal samples. Two hundred forty-nine strains (79%) were correctly serogrouped, 57 (18%) belonged to serogroups other than the 11 which were evaluated and gave a negative reaction with all antisera, and 8 isolates (2.5%) did not react with their corresponding antisera. ELISA is a rapid and reliable method for serogrouping C. difficile and should allow for the automation of this procedure. PMID- 8408582 TI - Severe human infections caused by Vibrio metschnikovii. AB - Vibrio metschnikovii is largely distributed in the aquatic environment; human infections are rarely observed. A fatal case of septicemia in a patient with liver cirrhosis, renal insufficiency, and diabetes is described. A second case in a 82-year-old woman with septicemia, respiratory problems, and infected leg lesions is reported; she was successfully treated. PMID- 8408583 TI - Ceftizoxime interpretive criteria for in vitro susceptibility tests with Neisseria gonorrhoeae. AB - Ceftizoxime was tested in triplicate against 100 clinical strains of Neisseria gonorrhoeae in accordance with National Committee for Clinical Laboratory Standards guidelines to establish susceptibility testing interpretive criteria. The MICs for 50 and 90% of the strains tested were 0.008 and 0.03 microgram/ml, respectively. These results confirm those of other studies reporting ceftizoxime's excellent activity against gonococci. Because no resistant strains were identified, a breakpoint MIC of < or = 0.5 microgram/ml was selected, with a correlate zone diameter of > or = 32 mm. Ceftizoxime appears to represent an alternative to other beta-lactamase-stable beta-lactams for cost-effective therapy of uncomplicated gonorrhea. PMID- 8408584 TI - Reliability of cord formation in BACTEC media for presumptive identification of mycobacteria. AB - We evaluated cord formation in BACTEC media as a criterion for presumptive identification of Mycobacterium tuberculosis complex. Auramine-rhodamine smears from 673 radiometrically positive BACTEC vials were examined. The presence of cording had a sensitivity, a specificity, and positive and negative predictive values of 22.9, 98.5, 82.9, and 80.3%, respectively. Before cord formation is used to report presumptive identification of M. tuberculosis complex, workers should determine the reliability of this finding in their own laboratories. PMID- 8408585 TI - Incidence of Neisseria gonorrhoeae isolates negative by Syva direct fluorescent antibody test but positive by Gen-Probe accuprobe test in a sexually transmitted disease clinic population. AB - To determine the accuracy of the Syva (Palo Alto, Calif.) direct fluorescent antibody (DFA) test in comparison with the Gen-Probe (San Diego, Calif.) Accuprobe culture confirmation test, we tested 395 isolates of Neisseria gonorrhoeae from cultures obtained from patients attending a sexually transmitted disease clinic from 1 July 1991 through 30 June 1992. All isolates were tested for DFA reactivity with a polyclonal reagent (Difco Laboratories, Detroit, Mich.) and a monoclonal reagent (Syva, Inc., direct specimen test) and for specific molecular probe reactivity by the Gen-Probe Accuprobe culture confirmation test for N. gonorrhoeae. The 395 isolates gave positive results for the Gen-Probe culture confirmation test and the Difco polyclonal direct specimen test. However, 18 (4.6%) of the isolates were negative for N. gonorrhoeae by the Syva DFA test. With the exception of six beta-lactamase-positive isolates, all isolates that were negative by Syva DFA were sensitive to penicillin, tetracycline, spectinomycin, and ceftriaxone by disk-diffusion susceptibility testing. Auxotyping and serotyping studies indicated that strains negative by Syva DFA consisted of several variants. The frequency of N. gonorrhoeae isolates showing negative results by Syva DFA in this patient population ranged from 0 to 11.5%/month. Laboratories using only the Syva DFA test for confirmation of N. gonorrhoeae may incur a significant risk of misidentification. PMID- 8408586 TI - Quality control guidelines for cefdinir, cefepime, cefetamet, cefmetazole, cefpodoxime, cefprozil, and clinafloxacin (CI-960) for various National Committee for Clinical Laboratory Standards susceptibility testing methods. Quality Control Study Group. AB - Several multilaboratory studies to determine quality control (QC) ranges for a variety of National Committee for Clinical Laboratory Standards (NCCLS) susceptibility tests are summarized. Replicate testing used multiple lots of media and antimicrobial disks in accordance with NCCLS recommendations, including the appropriate medium modifications for tests with Haemophilus spp. and Neisseria gonorrhoeae. QC ranges for MIC and disk diffusion testing of N. gonorrhoeae ATCC 49226 were proposed for cefepime, cefetamet, cefmetazole, and cefpodoxime. Disk diffusion QC ranges for Haemophilus influenzae ATCC 49247 or ATCC 49766 were recommended with cefepime, cefetamet (10- and 30-microgram disks), cefmetazole, cefpodoxime, and cefprozil. Disk diffusion QC ranges for Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 with cefdinir and clinafloxacin and those for Pseudomonas aeruginosa ATCC 27853 with clinafloxacin were also proposed. PMID- 8408587 TI - Recovery of a strain of Agrobacterium radiobacter with a mucoid phenotype from an immunocompromised child with bacteremia. AB - Agrobacteria are associated more commonly with plant than with human disease. The isolation of Agrobacterium radiobacter from blood cultures of an immunocompromised child with a transcutaneous catheter prompted a review of human infections caused by Agrobacterium species. Only 12 reports describing 19 cases of Agrobacterium infections in humans have appeared in the literature. Sixteen of the patients (84%) were equipped with implantable or transcutaneous medical devices at the time of infection, and 14 of the 19 (80%) patients could be considered immunocompromised because of underlying disease processes. Unlike those in previous reports, however, this patient was infected with a novel mucoid phenotype of A. radiobacter. Because of the significant relationship between infection and biomedical implants, we evaluated the adhesion of this mucoid strain and a nonmucoid strain of A. radiobacter to plastic by using two in vitro assays. No adhesion or biofilm formation was detected for either strain, but nonetheless it is clear from this review that the isolation of Agrobacterium spp. from patients with indwelling medical appliances should not be dismissed as an environmental contaminant. PMID- 8408588 TI - Human papillomavirus and cutaneous warts in meat handlers. AB - The association of papillomavirus and hand warts in meat handlers was examined. Human papillomavirus (HPV) DNA was found in 23 (88%) of 26 cutaneous warts, with HPV 7 (27%) and a yet unidentified HPV (HPV X) (42%) being the predominant types. HPV 2 was found in two (7.5%) patients, and HPV 4 was found in three (11.5%) patients. No bovine papillomavirus sequences were detected. In most patients, the warts developed in less than 2 years after they started working with meat. A possible HPV transmission route by protection gloves and professional equipment is suggested. PMID- 8408589 TI - Circulating Candida antigens and antibodies: useful markers of candidemia. AB - To investigate the utility of the 48-kDa antigen from Candida albicans in its commercial form (Directigen; Becton Dickinson) and three other serodiagnostic methods (detection of one antigen by Pastorex Candida [Sanofi Diagnostics Pasteur] and detection of immunoglobulin G [IgG] and IgM antibodies to C. albicans blastoconidia [bioMerieux]) for diagnosis of invasive Candida infection, we conducted a prospective clinical trial among 10 patients with candidemia (group 1), 30 patients colonized by C. albicans (group 2), 20 patients with bacteremia (group 3), and 20 subjects without clinical or microbiological evidence of infection. The Directigen system was positive for at least one serum sample each from eight patients in group 1. In groups 2, 3, and 4, it was positive for only three patients. There was no reaction to the Pastorex system in any of the patients infected with or colonized by C. albicans or in the non Candida-carrying controls. The IgG antibody concentration oscillated between 100 and 800 (mean, 510 +/- 268) IU/ml for the patients in group 1. In this group, eight patients had IgG antibody levels of > 400 IU/ml. The percentages of persons with IgG antibody levels of > 400 IU/ml in groups 2, 3, and 4 were 43.3, 0, and 0, respectively. Specific IgM antibody was present in all group 1 patients but not in those in groups 2, 3, and 4. The sensitivity and specificity of the Directigen test were 65 and 97.1%, respectively. For the Pastorex test, the sensitivity was 0%. The sensitivity of IgG antibodies was 80%, with a specificity of 81.4%, while the IgM antibodies were 100% specific and sensitive. Both the positive and negative predictive values of specific IgM antibodies appeared to be superior to those of the other three tests. PMID- 8408590 TI - Analysis of the humoral response to the flagellin protein of Borrelia burgdorferi. PMID- 8408591 TI - Use of selective buffered charcoal-yeast extract medium for isolation of nocardiae from mixed cultures. PMID- 8408592 TI - Rejection criteria for endotracheal aspirates. PMID- 8408593 TI - Scalp-recorded ictal patterns in focal epilepsy. AB - Scalp-recorded focal EEG seizure patterns are usually expressed as rhythmic metamorphic evolving patterns (with or without epileptiform morphology) that progress through two or more ictal phases into a postictal change. Such patterns are almost invariably seen in temporal complex partial seizures but less often detected in frontal complex partial seizures and least of all in simple partial seizures. The failure of scalp recordings to detect activity from a focal seizure can usually be explained by the seizure's distant location, limited extent, or disadvantageous orientation with respect to scalp electrodes. The elimination of these disadvantages with properly implanted electrodes explains why these recordings are able to detect seizure discharges missed by scalp electrodes. Although the lateralization of a scalp-recorded seizure can be misleading, it usually accurately identifies the focus when it remains well-lateralized throughout its various ictal phases and into the postictal state. PMID- 8408594 TI - Ictal patterns in generalized epilepsy. AB - Ictal EEG may be of great benefit in facilitating accurate classification of the underlying seizure disorder in some patients and thus guiding further investigation and management. Ictal recordings in patients with generalized epilepsies are protean in their manifestations and yet may have considerable overlap. Classification is only possible through careful synthesis of all available clinical and electrophysiological data. Although the underlying pathophysiological mechanisms of the generalized epilepsies remain uncertain, evidence from EEG recordings tends to support Gloor's concept of corticoreticular epilepsy. PMID- 8408595 TI - Ictal patterns in experimental models of epilepsy. AB - The goal of this article is to review many of the animal models that have been used for epilepsy research. The intent is to present the behavioral and electrographic patterns observed in these models and to relate those patterns to human epilepsy. This review is organized into three sections. The first section deals with the methods used in the study of epilepsy. These methods range from in vivo preparations, through in vitro brain slice preparations and neural grafting methods, to computer simulation methods. The second section of the review deals with agents capable of inducing epilepsy. Some of the agents include chemical convulsants, ionic changes in the tissue, changes in osmolarity, and withdrawal from drugs. The effects of these agents are reviewed with their relation to the mechanisms of action. Finally, the methods used to control epilepsy are reviewed. These methods include the application of anticonvulsants, changes in the osmolarity, neuronal grafts and electrical currents. Examples of epileptiform activity obtained from animal models published in the literature are shown in several figures for comparison to human patterns. It is clear from this review that animal models are capable of reproducing many of the seizure types found in clinical situations. Although we have some understanding of the mechanisms underlying the abnormal behavior in these animal models, we are unable to explain the mechanisms underlying human epilepsy. However, the combination of existing animal models, in vitro human tissue experiments, and computer stimulation should allow us to advance rapidly toward a solution. PMID- 8408596 TI - Development of clinical neurophysiology in the Chicago area. PMID- 8408597 TI - EEG and the neuropathology in premature neonates with intraventricular hemorrhage. AB - Eighty-eight EEGs from 32 premature neonates with autopsy-verified periventricular-intraventricular hemorrhage (PVH-IVH) were compared with associated neuropathological findings. PVH-IVH was rarely an isolated lesion at autopsy. Twenty-seven infants (84%) had additional parenchymal brain lesions, such as periventricular leukomalacia (47%), ischemic neuronal necrosis (22%), pontosubicular necrosis (22%), cerebral infarction (13%), and/or cerebellar hemorrhage (13%). A significant correlation was found between the patient's most abnormal EEG and the severity of morphological changes. Infants with more abnormal EEGs had more extensive brain lesions. On the other hand, IVH grade did not correlate with the degree of parenchymal brain lesions, with the exception of IVH with intraparenchymal involvement. Positive rolandic sharp wave transients (PRS) were observed in eight patients (25%). All infants with PRS had white matter lesions. The sensitivity of PRS for white matter lesions, however, was only 38%. EEG has limited value in the diagnosis of PVH-IVH or specific anatomical changes but provides useful electrographic information that correlates with the severity of brain damage in infants with PVH-IVH. PMID- 8408598 TI - The importance of source behavior in distinguishing populations of epileptic foci. AB - The interictal spike discharge is usually analyzed in terms of amplitude, morphology, location of spike negativity, frequency or pattern of occurrence, and the effect of sleep/wake cycle. Such information derived from the routine EEG supports the clinician in the diagnosis, treatment, and management of the epileptic patient. Detailed analytical or computerized studies of spike characteristics have not had a great deal of clinical impact to date. This communication presents an argument for characterizing the epileptic focus based on interictal spikes. Some findings from quantitative analysis of spike properties are reviewed. By facilitating the creation and refinement of hypotheses relating to the underlying neuronal mechanisms, mathematical source modeling (or dipole modeling) may enhance our understanding of the epileptic process. Such an approach is applied to rolandic epilepsy as an example, and a model is proposed that is parsimonious with experimental data and clinical observations. PMID- 8408599 TI - Prognostic value of background patterns in the neonatal EEG. AB - The prognostic value of background activity in the neonatal EEG has been well established. Whereas in older children the neonatal EEG is useful in the diagnosis of seizures, in neonates the test also provides a particularly valuable assessment of cerebral functioning following a variety of insults. In this review, the prognostic significance of abnormalities of amplitude, continuity, frequency, symmetry, synchrony, sleep state, and maturation are discussed. Certain abnormalities, such as cerebral electrical inactivity or burst suppression, are highly predictive of outcome, whereas other abnormalities of background activity are associated with more variable outcomes. Since the ill neonate may have more than one abnormality, predicting outcome based on a single EEG feature is discouraged. As in older children, drugs may affect EEG background rhythms. Although abnormalities on the neonatal EEG are not specific for diagnosis, certain EEG patterns may be highly suggestive for the diagnosis of pyridoxine dependency and neonatal herpes encephalitis. In both term and preterm infants, the prognostic value of the neonatal EEG is increased by performing serial studies. PMID- 8408600 TI - The use of a cap-shaped coil for transcranial magnetic stimulation of the motor cortex. AB - A cap-shaped coil is introduced as a superior design for inducing transcranial magnetic motor evoked potentials for spinal cord monitoring. Evaluation of the magnetic characteristics of the cap coil showed higher induced electrical fields at and below the depth of the cortical surface, compared to a 9-cm, butterfly shaped coil. Twenty normal adults were stimulated with the cap coil and a 9-cm round coil in three positions. Compound muscle action potentials were recorded from the left and right abductor digiti minimi and anterior tibialis muscles. The cap coil induced potentials with higher intensities and lower variability between consecutive stimuli. The cap coil was also more able to simultaneously induce motor evoked potentials from the four muscles studied. This coil design should provide superior means of inducing transcranial magnetic motor evoked potentials in multiple muscles. PMID- 8408601 TI - Monitoring carotid test occlusions with continuous EEG and clinical examination. AB - We routinely monitor invasive neuroradiologic carotid balloon test occlusions with continuous polygraph and quantitative EEG along with repeated detailed clinical examinations. Four of 17 consecutive cases showed changes during carotid occlusion. In one instance, an immediate delta increase was accompanied by slurred speech and aphasia. Another showed alpha attenuation without clinical change. A third patient had significant clinical change without EEG change. Nine of the 17 cases underwent permanent therapeutic carotid occlusion as treatment of an intracerebral vascular abnormality. Seven of these nine had no EEG or clinical changes during monitoring and have had no functional abnormalities on follow-up. The patient with focal alpha attenuation had an accidental balloon detachment but has had no functional or structural neurologic abnormalities. The patient with minor regional increased delta received a permanent carotid occlusion and went on to develop clinical signs 24 h later. We believe that continuous EEG monitoring and repeated clinical examinations provide useful ways of evaluating cerebral circulation during carotid test occlusions. PMID- 8408602 TI - Effects of acupuncture on somatosensory evoked potentials: a review. AB - Although acupuncture has a long history of analgesic effects, the mechanisms underlying its effects are still unclear. Somatosensory evoked potential (SEP) methodology has been adopted in the research of acupuncture since the 1970s. In research on the effects of acupuncture on the conventional SEP, variable results have been observed, and two different opinions concerning the presence or absence of acupuncture effects on the conventional SEP are discussed. Since the conventional SEP is mediated mainly by fast conducting sensory nerve fibers, the conventional SEP methodology, especially that for recording short-latency SEP, may be inadequate for studying acupuncture mechanisms. In the case of the long latency cortical SEP, there are too few data available to judge the effects of acupuncture analgesia (AA). Studies on the effects of AA on pain SEPs demonstrated that AA has a suppressive effect on amplitude of the pain SEP (and affecting the latency as well in some experiments) in both animals and humans, being accompanied by increased pain threshold. Thus, acupuncture seems to have analgesic effects that are probably related to activation of the antinociceptive system, and application of the pain SEP methodology to the study of mechanisms of AA may be promising. PMID- 8408603 TI - Technology and equipment review. Digital electroencephalographs. PMID- 8408604 TI - It's time for food irradiation. PMID- 8408605 TI - The Society for Epidemiologic Research and the future of epidemiology. AB - This paper describes the development of noninfectious disease epidemiology and the subsequent organization of the Society for Epidemiologic Research. It defines epidemiology as "the study of the health of human populations," and outlines its four basic functions. Certain negative trends in epidemiologic research are indicated. The current tasks of epidemiology are reviewed, as well as the objective and subjective factors which will determine the future of epidemiology. PMID- 8408606 TI - Public health aspects of food irradiation. AB - In view of the enormous health and economic consequences of foodborne diseases, the World Health Organization (WHO) encourages its Member States to consider all measures to eliminate or reduce foodborne pathogens in food and improve their supplies of safe and nutritious food. With the wholesomeness of irradiated food clearly established by extensive scientific studies, food irradiation has important roles to play in both ensuring food safety and reducing food losses. Food irradiation may be one of the most significant contributions to public health to be made by food science and technology since the introduction of pasteurization. Because the promotion of a safe, nutritious and adequate food supply is an essential component of its primary health care strategy, WHO is concerned that the unwarranted rejection of this process may endanger public health and deprive consumers of the choice of food processed for safety. PMID- 8408607 TI - The impact of sexism on women's health and health care. PMID- 8408608 TI - Reproductive rights and the medical care system: a plea for rational health policy. AB - Recently, there have been many challenges to women's reproductive rights and freedoms: court-ordered cesarean sections; criminal cases against women for prenatal child abuse; and attempts to limit the practice of mid-wifery, home birth, and the operation of alternative birth centers. In these cases, medicine has been complicit or proactive in attempts to control the behavior or health care options of pregnant women. We discuss medicine's role as an agent of social control, the medical reconstruction of problems that are social in nature, and the need for a more coherent policy framework to guide physician practices. PMID- 8408609 TI - Public health advocacy on behalf of women in Sao Paulo: learning to participate in the planning process. AB - In 1988, a new Constitution was adopted in Brazil in which guidelines for community participation in the development and implementation of the national health system were delineated. The health and welfare of women and children were given priority. Implementation of these guidelines presents a major challenge in a city such as Sao Paulo with a population of 15 million, of which an estimated 5.8 million are women of childbearing age. In order to determine the extent to which community organizations are actively participating in planning health services for women and children in Sao Paulo, a study was undertaken to examine the experience of community and professional organizations in public health advocacy. This paper describes a sample of these organizations, their constituents, membership, history, funding, advocacy objectives, and strategies used and results obtained. The information gathered indicates that the community organizations are involved in activities that include major efforts to improve access to health care by providing specialized courses in women's health, including the status of women's work, sexuality, discrimination, family planning, and the politics of health; publishing newsletters; producing radio programs; engaging in legal action; and using petitions, demonstrations, and public meetings to garner public support on specific issues. PMID- 8408610 TI - The reform of the Quebec health care system: potential for innovation? AB - The recent reform of the health care system in Quebec can be viewed as the result of a continuous process that originated with the first reform launched in the early 70s. The reform focuses on three elements: decentralization, citizen participation, and outcome-centered management. The context in which the reform is being launched contains both favorable conditions and obstacles to its successful implementation. PMID- 8408611 TI - The once and future health system in the former Yugoslavia: myths and realities. AB - This paper debunks three widely believed myths about the former Yugoslavia's health care system: that it was characterized by: (1) social ownership of "self managing" provider organizations; (2) a commitment to primary health care; and (3) a faith in what might be called the "march of progress"--the health system's continuous expansion and improvement. In contrast to this picture, we present an alternative view and conclude with a word of caution for American consultants and health care reformers in Eastern European countries and newly independent states: If universal health coverage is to be maintained, beware of reforms that do no more than substitute private for public organizational forms. PMID- 8408612 TI - Nurturing creativity: young Turks and young minds. PMID- 8408613 TI - Colchicine, crystals, and neutrophil tyrosine phosphorylation. PMID- 8408614 TI - Designer natriuretic peptides. PMID- 8408615 TI - Islet cell autoantigens in insulin-dependent diabetes. AB - A burgeoning number of antigenic targets of the islet cell autoimmunity in IDD have been identified, and more can be anticipated through improved methods for their identification. The challenge for those investigating the pathogenesis of IDD will be to assign the relative importance of these antigens to the development of the disease, and to resolve whether there is a dominant primary immunologic event that is followed by a series of secondary immunizations to a variety of normally sequestered islet cell antigens in the sequence of pathogenic events that culminate in IDD. One interesting observation that may have potential pathogenic implications is the observation that of all islet cell autoantigens described, only two (i.e., 64 kD/GAD, 38 kD) are reactive in their native configurations, implying that recognition of conformational epitopes is most important. This property argues for primary immunizing agents rather than secondary ones after release of denatured antigens and antigenic recognition through their epitopes. Given the complex and multiple physiological functions of islet cells and the continuous variation in their activity, it is reasonable to speculate that the speed of the progression to IDD could vary between individuals with respect to their insulin needs and the relative activities of their islets. Activated islets may express autoantigens that have only limited expression in quiescent islets. The often times striking variation in the severity of insulitis seen in different islets of a single pancreas may be explained by the level of activity of individual islets. Furthermore, disparity in HLA-DR/DQ associations with disease may involve differences in the immunological recognition of autoantigens. Whereas there is still much to learn, it is clear that disease predictability and disease intervention studies have been enhanced through the identification of the islet cell autoantigens in IDD. PMID- 8408616 TI - Demonstration of a critical role for free fatty acids in mediating counterregulatory stimulation of gluconeogenesis and suppression of glucose utilization in humans. AB - In vitro studies indicate that FFA compete with glucose as an oxidative fuel in muscle and, in addition, stimulate gluconeogenesis in liver. During counterregulation of hypoglycemia, plasma FFA increase and this is associated with an increase in glucose production and a suppression of glucose utilization. To test the hypothesis that FFA mediate changes in glucose metabolism that occur during counterregulation, we examined the effects of acipimox, an inhibitor of lipolysis, on glucose production and utilization ([3-3H]glucose), and incorporation of [U-14C]-alanine into glucose during insulin-induced hypoglycemia. Eight normal volunteers were infused with insulin for 8 h to produce modest hypoglycemia (approximately 3 mM) on two occasions, first without acipimox (control) and then with acipimox administration (250 mg per os at 60 and 240 min). Despite identical plasma insulin concentrations, glucose had to be infused in the acipimox experiments (glucose-clamp technique) to maintain plasma glucose concentrations identical to those in control experiments. Acipimox completely prevented counterregulatory increases in lipolysis so that during the last 4 h plasma FFA were below baseline values and averaged 67 +/- 13 vs. 725 +/- 65 microM in control experiments, P < 0.001. Concomitantly, overall glucose production was reduced by 40% (5.5 +/- 11 vs. 9.3 +/- 0.7 mumol/kg per min, P < 0.001), and gluconeogenesis from alanine was reduced by nearly 70% (0.32 +/- 0.09 vs. 1.00 +/- 0.18 mumol/kg per min, P < 0.001), while glucose utilization increased by 15% (10.8 +/- 1.4 vs. 9.3 +/- 0.7 mumol/kg per min). We conclude that FFA play a critical role in mediating changes in glucose metabolism during counterregulation, and that under these conditions, FFA exert a much more profound effect on hepatic glucose production than on glucose utilization. PMID- 8408617 TI - Mechanism of enhanced insulin sensitivity in athletes. Increased blood flow, muscle glucose transport protein (GLUT-4) concentration, and glycogen synthase activity. AB - We examined the mechanisms of enhanced insulin sensitivity in 9 male healthy athletes (age, 25 +/- 1 yr; maximal aerobic power [VO2max], 57.6 +/- 1.0 ml/kg per min) as compared with 10 sedentary control subjects (age, 28 +/- 2 yr; VO2max, 44.1 +/- 2.3 ml/kg per min). In the athletes, whole body glucose disposal (240-min insulin clamp) was 32% (P < 0.01) and nonoxidative glucose disposal (indirect calorimetry) was 62% higher (P < 0.01) than in the controls. Muscle glycogen content increased by 39% in the athletes (P < 0.05) but did not change in the controls during insulin clamp. VO2max correlated with whole body (r = 0.60, P < 0.01) and nonoxidative glucose disposal (r = 0.64, P < 0.001). In the athletes forearm blood flow was 64% greater (P < 0.05) than in the controls, whereas their muscle capillary density was normal. Basal blood flow was related to VO2max (r = 0.63, P < 0.05) and glucose disposal during insulin infusion (r = 0.65, P < 0.05). The forearm glucose uptake in the athletes was increased by 3.3 fold (P < 0.01) in the basal state and by 73% (P < 0.05) during insulin infusion. Muscle glucose transport protein (GLUT-4) concentration was 93% greater in the athletes than controls (P < 0.01) and it was related to VO2max (r = 0.61, P < 0.01) and to whole body glucose disposal (r = 0.60, P < 0.01). Muscle glycogen synthase activity was 33% greater in the athletes than in the controls (P < 0.05), and the basal glycogen synthase fractional activity was closely related to blood flow (r = 0.88, P < 0.001). IN CONCLUSION: (a) athletes are characterized by enhanced muscle blood flow and glucose uptake. (b) The cellular mechanisms of glucose uptake are increased GLUT-4 protein content, glycogen synthase activity, and glucose storage as glycogen. (c) A close correlation between glycogen synthase fractional activity and blood flow suggests that they are causally related in promoting glucose disposal. PMID- 8408618 TI - Delayed catabolism of high density lipoprotein apolipoproteins A-I and A-II in human cholesteryl ester transfer protein deficiency. AB - Deficiency of the cholesteryl ester transfer protein (CETP) in humans is characterized by markedly elevated plasma concentrations of HDL cholesterol and apoA-I. To assess the metabolism of HDL apolipoproteins in CETP deficiency, in vivo apolipoprotein kinetic studies were performed using endogenous and exogenous labeling techniques in two unrelated homozygotes with CETP deficiency, one heterozygote, and four control subjects. All study subjects were administered 13C6-labeled phenylalanine by primed constant infusion for up to 16 h. The fractional synthetic rates (FSRs) of apoA-I in two homozygotes with CETP deficiency (0.135, 0.134/d) were found to be significantly lower than those in controls (0.196 +/- 0.041/d, P < 0.01). Delayed apoA-I catabolism was confirmed by an exogenous radiotracer study in one CETP-deficient homozygote, in whom the fractional catabolic rate of 125I-apoA-I was 0.139/d (normal 0.216 +/- 0.018/d). The FSRs of apoA-II were also significantly lower in the homozygous CETP deficient subjects (0.104, 0.112/d) than in the controls (0.170 +/- 0.023/d, P < 0.01). The production rates of apoA-I and apoA-II were normal in both homozygous CETP-deficient subjects. The turnover of apoA-I and apoA-II was substantially slower in both HDL2 and HDL3 in the CETP-deficient homozygotes than in controls. The kinetics of apoA-I and apoA-II in the CETP-deficient heterozygote were not different from those in controls. These data establish that homozygous CETP deficiency causes markedly delayed catabolism of apoA-I and apoA-II without affecting the production rates of these apolipoproteins. PMID- 8408619 TI - Cytokine control of parasite-specific anergy in human lymphatic filariasis. Preferential induction of a regulatory T helper type 2 lymphocyte subset. AB - The immunological mechanisms involved in maintenance of an asymptomatic microfilaremic state (MF) in patients with lymphatic filariasis remain undefined. MF patients have impaired filarial antigen (Ag)-specific lymphocyte proliferation and decreased frequencies (Fo) of Ag-specific T cells, and yet elevated serum IgE and antifilarial IgG4. To investigate the mechanism of Ag-specific anergy in MF patients in contrast to amicrofilaremic individuals with chronic lymphatic obstruction (CP), the Fo of Ag-specific lymphocytes from peripheral blood mononuclear cells secreting either IL-4 or IFN-gamma were assessed by filter spot enzyme-linked immunosorbent assay, and IL-10 and transforming growth factor-beta (TGF-beta) mRNA transcript levels were assessed by a semiquantitative reverse transcriptase polymerase chain reaction technique. The Fo of filaria-specific IL 4-secreting lymphocytes were equivalent in both MF (geometric mean [GM] = 1:11,700) and CP (GM = 1:29,300 P = 0.08), whereas the Fo of IFN-gamma-secreting lymphocytes were lower in MF (GM = 1:39,300) than in CP (GM = 1:4,200, P < 0.01). When the ratio of IL-4/IFN-gamma (T helper type 2 [Th2]/Th1)-secreting cells was examined, MF subjects showed a predominant Th2 response (8:1) compared with a Th1 response in CP individuals (1:4). mRNA transcript levels of IL-10 were also significantly elevated in MF compared with CP individuals (P < 0.01). Further, IL 10 and TGF-beta were shown to have a role in modulating the Ag-specific anergy among MF subjects, in that neutralizing anti-IL-10 or anti-TGF-beta significantly enhanced lymphocyte proliferation response (by 220-1,300%) to filarial Ags in MF individuals. These findings demonstrate that MF subjects respond to parasite antigen by producing a set of suppressive cytokines that may facilitate persistence of the parasite within humans while producing little clinical disease. PMID- 8408620 TI - Cultured human liver fat-storing cells produce monocyte chemotactic protein-1. Regulation by proinflammatory cytokines. AB - Monocytes infiltrate the portal space during chronic liver inflammation. Monocyte chemotactic protein-1 (MCP-1) is a cytokine that induces monocyte chemotaxis and activation. We investigated if human liver fat-storing cells (FSC) secrete MCP-1, and the mechanisms that regulate MCP-1 production. Unstimulated FSC secrete MCP-1 as measured by radioimmunoassay as well as a chemotactic assay and express mRNA that encodes for this cytokine. A two- to threefold increase in MCP-1 secretion was observed when FSC were treated with either interleukin-1 alpha (IL-1 alpha) or interferon-gamma (IFN-gamma). Tumor necrosis factor-alpha (TNF alpha) also increased MCP-1 secretion, although to a lesser extent (1.6-fold). Northern blot analysis showed that IL-1 alpha and IFN-gamma strongly increase the levels of mRNA that encodes for MCP-1, whereas TNF alpha appears to be a weaker stimulus. Analysis of FSC-conditioned medium by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting revealed three bands of MCP-1 that most likely represent isoforms of different apparent molecular weights. Pretreatment of FSC with H-7, a protein kinase C inhibitor, blocked cytokine-induced increase in both MCP-1 gene expression and secretion. To determine the potential role of MCP-1 in vivo, we also analyzed normal and pathologic human liver tissue. Northern blot analysis showed that MCP-1 mRNA expression is more abundant in liver tissue obtained from patients with chronic active hepatitis compared with normal liver tissue. These studies indicate that MCP-1 secreted by FSC is stimulated by proinflammatory cytokines and that MCP-1 gene expression is upregulated in chronic inflammatory liver disease. MCP-1 released by FSC may participate in the recruitment and activation of monocytes at sites of liver injury. PMID- 8408621 TI - Definition of immunoglobulin A receptors on eosinophils and their enhanced expression in allergic individuals. AB - Fc alpha receptors (Fc alpha R), detected by the binding of IgA and by anti-Fc alpha R antibodies, were found to be differentially expressed on eosinophils and neutrophils. Neutrophils were the major granulocyte population expressing Fc alpha R, and they expressed much higher levels of Fc alpha R than eosinophils. The expression of Fc alpha R by eosinophils could be upregulated approximately threefold by Ca2+ ionophore treatment in a dose- and time-dependent manner. This effect, which was blocked by a chelating agent, was not duplicated by other cellular stimuli. Eosinophils in allergic individuals displayed enhanced Fc alpha R expression, whereas neutrophils did not. The Fc alpha R on eosinophils had a higher molecular mass (70-100 kD) than those identified on neutrophils (55-75 kD). However, removal of N-linked carbohydrates from Fc alpha R of eosinophils and neutrophils revealed a major protein core of 32 kD for both cell types. The data indicate that expression of Fc alpha R molecules with a characteristic glycosylation pattern is upregulated on eosinophils in allergic individuals. PMID- 8408622 TI - Osteopontin is elevated during neointima formation in rat arteries and is a novel component of human atherosclerotic plaques. AB - In an earlier report, we used differential cloning to identify genes that might be critical in controlling arterial neointima formation (Giachelli, C., N. Bae, D. Lombardi, M. Majesky, and S. Schwartz. 1991. Biochem. Biophys. Res. Commun. 177:867-873). In this study, we sequenced the complete cDNA and conclusively identified one of these genes, 2B7, as rat osteopontin. Using immunochemistry and in situ hybridization, we found that medial smooth muscle cells (SMC) in uninjured arteries contained very low levels of osteopontin protein and mRNA. Injury to either the adult rat aorta or carotid artery using a balloon catheter initiated a qualitatively similar time-dependent increase in both osteopontin protein and mRNA in arterial SMC. Expression was transient and highly localized to neointimal SMC during the proliferative and migratory phases of arterial injury, suggesting a possible role for osteopontin in these processes. In vitro, basic fibroblast growth factor (bFGF), transforming growth factor-beta (TGF beta), and angiotensin II (AII), all proteins implicated in the rat arterial injury response, elevated osteopontin expression in confluent vascular SMC. Finally, we found that osteopontin was a novel component of the human atherosclerotic plaque found most strikingly associated with calcified deposits. These data implicate osteopontin as a potentially important mediator of arterial neointima formation as well as dystrophic calcification that often accompanies this process. PMID- 8408623 TI - A murine skeletal adaptation that significantly increases cortical bone mechanical properties. Implications for human skeletal fragility. AB - Mov13 mice carry a provirus that prevents transcription initiation of the alpha 1(I) collagen gene. Mutant mice homozygous for the null mutation produce no type I collagen and die at mid-gestation, whereas heterozygotes survive to adulthood. Dermal fibroblasts from heterozygous mice produce approximately 50% less type I collagen than normal littermates, and the partial deficiency in collagen production results in a phenotype similar to osteogenesis imperfecta type I (an inherited form of skeletal fragility). In this study, we have identified an adaptation of Mov13 skeletal tissue that significantly improves the bending strength of long bone. The adaptive response occurred over a 2-mo period, during which time a small number of newly proliferated osteogenic cells produced a significant amount of matrix components and thus generated new bone along periosteal surfaces. New bone deposition resulted in a measurable increase in cross-sectional geometry which, in turn, led to a dramatic increase in long bone bending strength. PMID- 8408624 TI - Disruption of cytoskeletal structures mediates shear stress-induced endothelin-1 gene expression in cultured porcine aortic endothelial cells. AB - Hemodynamic shear stress alters the architecture and functions of vascular endothelial cells. We have previously shown that the synthesis of endothelin-1 (ET-1) in endothelial cells is increased by exposure to shear stress. Here we examined whether shear stress-induced alterations in cytoskeletal structures are responsible for increases in ET-1 synthesis in cultured porcine aortic endothelial cells. Exposure of endothelial cells to 5 dyn/cm2 of low shear stress rapidly increased monomeric G-actin contents within 5 min without changing total actin contents. The ratio of G- to total actin, 54 +/- 0.8% in quiescent endothelial cells, increased to 87 +/- 4.2% at 6 h and then decreased. Following the disruption of filamentous (F)-actin into G-actin, ET-1 mRNA levels in endothelial cells also increased within 30 min and reached a peak at 6 h. The F actin stabilizer, phalloidin, abolished shear stress-induced increases in ET-1 mRNA; however, it failed to inhibit increases in ET-1 mRNA secondary to other stimulants. This indicates that shear stress-induced increases in ET-1 mRNA levels may be mediated by the disruption of actin fibers. Furthermore, increases in ET-1 gene expression can be induced by actin-disrupting agents, cytochalasin B and D. Another cytoskeleton-disrupting agent, colchicine, which inhibits dimerization of tubulin, did not affect the basal level of ET-1 mRNA. However, colchicine completely inhibited shear stress- and cytochalasin B-induced increases in ET-1 mRNA levels. These results suggest that shear stress-induced ET 1 gene expression in endothelial cells is mediated by the disruption of actin cytoskeleton and this induction is dependent on the integrity of microtubules. PMID- 8408625 TI - Reduced sympathetic nervous activity. A potential mechanism predisposing to body weight gain. AB - The sympathetic nervous system is recognized to play a role in the etiology of animal and possibly human obesity through its impact on energy expenditure and/or food intake. We, therefore, measured fasting muscle sympathetic nerve activity (MSNA) in the peroneal nerve and its relationship with energy expenditure and body composition in 25 relatively lean Pima Indian males (means +/- SD; 26 +/- 6 yr, 82 +/- 19 kg, 28 +/- 10% body fat) and 19 Caucasian males (29 +/- 5 yr, 81 +/ 13 kg, 24 +/- 9% body fat). 24-h energy expenditure, sleeping metabolic rate, and resting metabolic rate were measured in a respiratory chamber, whereas body composition was estimated by hydrodensitometry. Pima Indians had lower MSNA than Caucasians (23 +/- 6 vs 33 +/- 10 bursts/min, P = 0.0007). MSNA was significantly related to percent body fat in Caucasians (r = 0.55, P = 0.01) but not in Pimas. MSNA also correlated with energy expenditure adjusted for fat-free mass, fat mass, and age in Caucasians (r = 0.51, P = 0.03; r = 0.54, P = 0.02; and r = 0.53, P = 0.02 for adjusted 24-h energy expenditure, sleeping metabolic rate, and resting metabolic rate, respectively) but not in Pima Indians. In conclusion, the activity of the sympathetic nervous system is a determinant of energy expenditure in Caucasians. Individuals with low resting MSNA may be at risk for body weight gain resulting from a lower metabolic rate. A low resting MSNA and the lack of impact of MSNA on metabolic rate might play a role in the etiology of obesity in Pima Indians. PMID- 8408626 TI - Human monocyte colony-stimulating factor stimulates the gene expression of monocyte chemotactic protein-1 and increases the adhesion of monocytes to endothelial monolayers. AB - The stimulation of the human umbilical vein endothelial cell (HUVEC) with recombinant human monocyte-derived colony-stimulating factor (MCSF) increased the gene expression of monocyte chemotactic protein (MCP-1). Northern blot analysis indicated that 50 U/ml of MCSF is the optimal concentration for this effect. The elevation of MCP-1 mRNA started as early as 1 h after stimulation and was maintained for at least 8 h. An increased MCP-1 level in MCSF-treated HUVEC was also demonstrated at the protein level by immunocytochemical staining using a polyclonal MCP-1-specific antibody. HUVEC activated by 50 U/ml of MCSF for 5 h showed a stronger immunofluorescence staining than control cells. Micropipette separation of THP-1 monocytes from HUVEC showed that the activation of both THP-1 and endothelium by MCSF led to an increase in the separation force by more than three times (36.2 +/- 6.7 x 10(-4) vs. 9.6 +/- 3.6 x 10(-4) dyn). An increased adhesiveness was also observed after MCSF activation of peripheral blood monocytes and HUVEC (16.7 +/- 2.7 x 10(-4) vs. 5.2 +/- 0.9 x 10(-4) dyn). The increased adhesive force in both systems was blocked by the use of anti-MCP-1 (5.5 +/- 0.8 x 10(-4) and 6.8 +/- 1.1 x 10(-4) dyn). Similar results were obtained in experiments in which only HUVEC, but not monocytes, were activated by MCSF. This increased adhesion of untreated monocytes to MCSF-activated HUVEC was also blocked by the addition of anti-MCP-1. In contrast, experiments in which only THP-1 or peripheral blood monocytes, but not HUVEC, were treated with MCSF did not show a significant increase of adhesion between these cells. These results indicate that MCSF augments monocyte-endothelium interaction primarily by its action on the endothelial cell and that this function is probably mediated through an increased expression of MCP-1. The MCSF/MCP-1-dependent adhesive mechanism might be operative in the arterial wall in vivo to lead to the trapping of the infiltrated monocyte-macrophage in the subendothelial space during atherogenesis. PMID- 8408627 TI - Intravenous glucose suppresses glucose production but not proteolysis in extremely premature newborns. AB - To ascertain whether the inability to suppress glucose production and increase glucose utilization in response to glucose infusion is an inherent characteristic of immature individuals, we determined glucose rate of appearance (R(a)) in minimally stressed, clinically stable, extremely premature infants (approximately 26-wk gestation) at two glucose infusion rates (6.2 +/- 0.4 and 9.5 +/- 0.5 mg/kg per min). We also assessed whether an increase in glucose delivery suppresses proteolysis by measuring the R(a) of phenylalanine and leucine. Glucose R(a) (and utilization) increased significantly at the higher glucose infusion rate (7.9 +/- 0.5 vs. 9.8 +/- 0.6 mg/kg per min). Glucose production persisted at the lower glucose infusion rate but was suppressed to nearly zero at the higher rate (1.7 +/- 0.5 vs. 0.3 +/- 0.1 mg/kg per min). Proteolysis was unaffected by the higher glucose infusion rate as reflected by no change in the rates of appearance of either phenylalanine (96 +/- 5 vs. 95 +/- 3 mumol/kg per h) or leucine (285 +/- 20 vs. 283 +/- 14 mumol/kg per h). Thus, clinically stable, extremely premature infants suppress glucose production and increase glucose utilization in response to increased glucose infusion, demonstrating no inherent immaturity of these processes. In contrast, increasing the rate of glucose delivery results in no change in whole body proteolysis in these infants. The regulation of proteolysis in this population remains to be defined. PMID- 8408628 TI - Expression of lipoprotein lipase mRNA and secretion in macrophages isolated from human atherosclerotic aorta. AB - The expression of lipoprotein lipase (LPL) mRNA and the LPL activity were studied in macrophages (CD14 positive) from human atherosclerotic tissue. Macrophages were isolated after collagenase digestion by immunomagnetic isolation. About 90% of the cells were foam cells with oil red O positive lipid droplets. To analyze the mRNA expression, PCR with specific primers for LPL was used. Arterial macrophages were analyzed directly after isolation and the data showed low expression of LPL mRNA when compared with monocyte-derived macrophages. To induce the expression of LPL mRNA in macrophages, PMA was used. When incubating arterial macrophages with PMA for 24 h we could not detect any increase in LPL mRNA levels. Similarly, the cells secreted very small amounts of LPL even after PMA stimulation. In conclusion, these studies show a very low expression of LPL mRNA in the CD14-positive macrophage-derived foam cells isolated from human atherosclerotic tissue. These data suggest that the CD14-positive cells are a subpopulation of foam cells that express low levels of lipoprotein lipase, and the lipid content could be a major factor for downregulation of LPL. However, the cells were isolated from advanced atherosclerotic lesions, and these findings may not reflect the situation in early fatty streaks. PMID- 8408629 TI - Altered regulation of apolipoprotein A-IV gene expression in the liver of the genetically obese Zucker rat. AB - Apolipoprotein (apo) A-IV, a structural component of chylomicrons and high density lipoproteins, may play a role in the catabolism of triglyceride-rich lipoproteins and in reverse cholesterol transport. To study the regulation of apoA-IV gene expression by genetic and nutritional factors, we determined the effect of a fish oil-rich and a sucrose-rich diet on apoA-IV gene transcription and nuclear and total cellular apoA-IV mRNA abundance in livers of genetically obese, hyperlipoproteinemic (fa/fa) Zucker rats and their lean (Fa/-) littermates. In obese rats fed chow, hepatic apoA-IV gene expression was more than twofold higher than in lean rats because of a post-transcriptional mechanism. apoA-I gene expression and apoC-III mRNA levels, studied as controls, were similar in both groups. The fish oil-rich diet reduced total cellular apoA IV mRNA abundance transcriptionally to 34 +/- 4% of basal values in lean rats, but did not alter apoA-IV gene expression in obese rats. In contrast, this diet reduced apoA-I gene expression in both lean and obese animals. The sucrose-rich diet increased apoA-IV gene expression twofold in both lean and obese rats. Thus, genetic obesity alters the response of hepatic apoA-IV gene expression to a lipid lowering diet rich in fish oil by a mechanism affecting transcriptional regulation. PMID- 8408630 TI - Expression of muscle-type phosphorylase in innervated and aneural cultured muscle of patients with myophosphorylase deficiency. AB - Patients with McArdle's myopathy lack muscle glycogen phosphorylase (M-GP) activity. Regenerating and cultured muscle of patients with McArdle's myopathy presents a glycogen phosphorylase (GP) activity, but it is not firmly established whether M-GP or non-M-GP isoforms are expressed. We have cultured myoblasts from biopsy specimen of five patients with McArdle's myopathy. Skeletal muscle was cultured aneurally or was innervated by coculture with fetal rat spinal cord explants. In the patients' muscle biopsies and in their cultured innervated and aneural muscle we studied total GP activity, isoenzymatic pattern, reactivity with anti-M-GP antiserum, and presence of M-GP mRNA. There was no detectable enzymatic activity, no immunoreactivity with anti-M-GP antiserum, and no M-GP mRNA in the muscle biopsy of all patients. GP activity, M-GP isozyme, and anti-M GP antiserum reactivity were present in patients' aneural cultures, increased after innervation, and were undistinguishable from control. M-GP mRNA was demonstrated in both aneural and innervated cultures of patients and control by primer extension and PCR amplification of total RNA. Our studies indicate that the M-GP gene is normally transcribed and translated in cultured muscle of patients with myophosphorylase deficiency. PMID- 8408631 TI - CAMP-mediated inhibition of the renal brush border membrane Na+-H+ exchanger requires a dissociable phosphoprotein cofactor. AB - Prior studies have suggested that protein kinase A (PKA)-mediated inhibition of the rabbit renal brush border membrane (BBM) Na(+)-H+ exchanger involves a regulatory protein that is distinct from the transporter. This putative regulatory protein was purified by column chromatography and SDS-PAGE, and a partial primary amino acid sequence was determined. An affinity-purified polyclonal antibody to a synthetic peptide representing a sequence of the protein recognized a polypeptide of 55 kD in BBM but not in basolateral membrane. The antibody immunoprecipitated a PKA substrate of a similar molecular mass from detergent-solubilized BBM proteins. When assayed after reconstitution, PKA in the presence of ATP and Mg2+ did not inhibit Na(+)-H+ exchange transport in a fraction of solubilized BBM proteins eluting from an anion exchange column between 0.2 and 0.4 M NaCl (fraction B). Coreconstitution of fraction B with the immunoprecipitated 55-kD protein restored the inhibitory effect of PKA (change = 42%, P < 0.05). By contrast, Na(+)-H+ exchange transport in total solubilized BBM proteins was inhibited 25% (P < 0.05) by PKA, ATP, and Mg2+. This effect was abolished by immunodepletion of the cAMP regulatory protein (change = +5%, P = NS). These findings provide evidence that the regulation of renal BBM Na(+)-H+ exchange transport by PKA is affected by repletion and depletion of a specific protein. This suggests that PKA-mediated inhibition of the renal BBM Na(+)-H+ exchanger requires participation of a regulatory protein that is distinct from the transporter itself. PMID- 8408632 TI - Activation-dependent contractility of rat hepatic lipocytes in culture and in vivo. AB - Hepatic lipocytes are perisinusoidal cells that have been thought to be analogous to tissue pericytes, a cell type with purported vasoregulatory properties. However, we and others have recently demonstrated that lipocytes acquire markers of smooth muscle cells or myofibroblasts only after liver injury, via a process termed "activation." In this study, we document lipocyte contractility on collagen lattices and examine the importance of activation in this process. In culture, lipocytes became contractile only after spreading and activating, coincident with expression of smooth muscle alpha actin, a marker of activation (1990. Virchows Arch. B Cell Pathol. 59:349). After 5 d in culture, lipocytes induced rapid and sustained contraction of collagen lattices (to 43.7 +/- 2.3% of their original size 24 h after detachment). There was no contraction of lattices containing hepatocytes. Scanning electron microscopy demonstrated intimate associations of lipocyte cell membranes and collagen fibrils. Reduction in cell volume during contraction was also prominent. Lattice contraction by lipocytes was proportional to cell number. Serum was a potent stimulator of lipocyte contraction, as were endothelin types 1, 2, and 3; the effect of serum and endothelin 1 were additive. Neither thrombin, angiotensin-II, serotonin, nor the cytokines PDGF and TGF beta induced contraction. Cytochalasin B treatment resulted in concentration-dependent inhibition of contraction. As a test of the in vivo relevance of the culture findings, lipocytes were isolated from fibrotic animals and examined immediately after adherence. Whereas lipocytes from normal liver were initially compact, smooth muscle alpha actin negative and noncontractile, cells from animals with hepatic injury due to CCl4 displayed an activated appearance, expressed smooth muscle alpha actin, and were contractile immediately after adherence. Additionally, IFN-gamma, an agent which blocks lipocyte activation (1992. Hepatology. 16:776), inhibited lipocyte contraction. The data document that normal (i.e., quiescent) lipocytes are not contractile, but that activation is associated with the development of contractility. These findings suggest that a role for lipocytes in organ contraction or vasoregulation may be confined to injured, not normal liver. PMID- 8408633 TI - Distinct nuclear proteins competing for an overlapping sequence of cyclic adenosine monophosphate and negative regulatory elements regulate tissue-specific mouse renin gene expression. AB - The mouse renin locus (Ren-1d) exhibits specific patterns of tissue expression. It is expressed in kidney but not submandibular gland (SMG). This locus contains a negative regulatory element (NRE) and a cAMP responsive element (CRE) that share an overlapping sequence. In the kidney, CRE binding proteins (CREB) and NRE binding proteins (NREB) compete for binding to this sequence, with the CREB having a greater affinity. In the SMG, CREB is inactivated by an inhibitory protein, permitting NREB to bind to the sequence, thus inhibiting Ren-1d expression. We hypothesize that the competition between NREB and CREB for this sequence governs tissue-specific expression of mouse renin. We speculate that this may be a general paradigm that determines tissue-specific gene expression. PMID- 8408634 TI - Effects of psychosocial stress on endothelium-mediated dilation of atherosclerotic arteries in cynomolgus monkeys. AB - The objectives of this study were to determine if psychosocial stress impairs dilation through endothelium-derived relaxing factor (EDRF)-mediated mechanisms and if this effect is long lasting. Monkeys were fed an atherogenic diet for 36 mo while in one of three experimental conditions: (a) stable social groups ("unstressed," n = 6); (b) unstable social groups for the first half of the experiment and stable groups for the second half ("early stress," n = 8); and (c) stable groups for the first half of the experiment and unstable groups for the second half ("late stress," n = 6). Iliac arteries were studied in organ chambers containing Krebs' buffer and 10(-6) M indomethacin. Arteries from the late stress group had impaired dilation (shift of the dose-response curve down and to the right) to acetylcholine and the calcium ionophore A23187 (for both, P < 0.05), but not to nitroprusside (P > 0.05), compared with unstressed or early stress monkeys. NG-methyl-L-arginine reduced the dose-response curve to both acetylcholine and A23187 in the unstressed group and resulted in similar vascular responses among all three groups (P > 0.05). We conclude that current, but not previous, exposure to chronic stress impairs endothelium-mediated dilation of atherosclerotic iliac arteries of cynomolgus monkeys through an EDRF-mediated mechanism. PMID- 8408635 TI - Saturable transport of insulin from plasma into the central nervous system of dogs in vivo. A mechanism for regulated insulin delivery to the brain. AB - By acting in the central nervous system, circulating insulin may regulate food intake and body weight. We have previously shown that the kinetics of insulin uptake from plasma into cerebrospinal fluid (CSF) can best be explained by passage through an intermediate compartment. To determine if transport kinetics into this compartment were consistent with an insulin receptor-mediated transport process, we subjected overnight fasted, anesthetized dogs to euglycemic intravenous insulin infusions for 90 min over a wide range of plasma insulin levels (69-5,064 microU/ml) (n = 10). Plasma and CSF samples were collected over 8 h for determination of immunoreactive insulin levels, and the kinetics of insulin uptake from plasma into CSF were analyzed using a compartmental model with three components (plasma-->intermediate compartment-->CSF). By sampling frequently during rapid changes of plasma and CSF insulin levels, we were able to precisely estimate three parameters (average standard deviation 14%) characterizing the uptake of insulin from plasma, through the intermediate compartment and into CSF (k1k2); insulin entry into CSF and insulin clearance from the intermediate compartment (k2 + k3); and insulin clearance from CSF (k4). At physiologic plasma insulin levels (80 +/- 7.4 microU/ml), k1k2 was determined to be 10.7 x 10(-6) +/- 1.3 x 10(-6) min-2. With increasing plasma levels, however, k1k2 decreased progressively, being reduced sevenfold at supraphysiologic levels (5,064 microU/ml). The apparent KM of this saturation curve was 742 microU/ml (approximately 5 nM). In contrast, the rate constants for insulin removal from the intermediate compartment and from CSF did not vary with plasma insulin (k2 + k3 = 0.011 +/- 0.0019 min-1 and k4 = 0.046 +/- 0.021 min-1). We conclude that delivery of plasma insulin into the central nervous system is saturable, and is likely facilitated by an insulin-receptor mediated transport process. PMID- 8408636 TI - Distinct mechanisms of epithelial adhesion for Candida albicans and Candida tropicalis. Identification of the participating ligands and development of inhibitory peptides. AB - The yeast Candida albicans is the leading cause of disseminated fungal infection in neonates, immunocompromised hosts, diabetics, and postoperative patients; Candida tropicalis is the second most frequent isolate. Because the integrin analogue in C. albicans shares antigenic, structural, and functional homologies with the beta 2-integrin subunits alpha M and alpha X, we investigated the role of integrin analogues in epithelial adhesion of C. albicans and C. tropicalis. On flow cytometry with the monoclonal antibody (mAb) OKM1, surface fluorescence was highest for C. albicans and significantly reduced for C. tropicalis (P < 0.001). However, adhesion to the human epithelial cell line HeLa S3 did not differ for these two candidal species: specific adhesion was highest for C. albicans at 44.0 +/- 1.8%, and only slightly lower for C. tropicalis at 38.8 +/- 3.6% (P = NS). The disparity between expression of the integrin analogue and epithelial adhesion suggested distinct mechanisms for this process in C. albicans versus C. tropicalis. Preincubation of C. albicans with anti-alpha M mAbs, with purified iC3b (the RGD ligand for the integrin analogue), or with 9-15-mer RGD peptides from iC3b all inhibited epithelial adhesion significantly (P < 0.001-0.04). Purified fibronectin or fibronectin-RGD peptides failed to block C. albicans adhesion. In contrast, epithelial adhesion of C. tropicalis was significantly inhibited by purified fibronectin and its RGD peptides (P < or = 0.021), but not by iC3b nor the iC3b-RGD peptides. Both iC3b and fibronectin were identified on the surface of epithelial cells after growth in serum-free medium. A polyclonal antibody to C3 inhibited C. albicans adhesion while a control antibody to fibronectin was ineffective; the converse was true for C. tropicalis. These results indicate that the pathogenic yeasts C. albicans and C. tropicalis recognize distinct RGD ligands present at the surface of the epithelial cell and that these interactions can be differentially inhibited by defined RGD peptides containing appropriate flanking sequences. PMID- 8408637 TI - Upregulation of fibronectin synthesis by interleukin-1 beta in coronary artery smooth muscle cells is associated with the development of the post-cardiac transplant arteriopathy in piglets. AB - We previously described in piglets after heterotopic cardiac transplantation the early development of a coronary arteriopathy characterized by increased immunostaining for fibronectin and interleukin-1 beta (IL-1 beta) in the vessel wall. The objective of this study was to culture smooth muscle cells from donor and host coronary arteries in these piglets to determine whether donor cells produce more fibronectin than host cells as judged by increased protein and mRNA levels, and whether IL-1 beta may be regulating this increase by an autocrine mechanism involving increased production of the cytokine. We documented increased donor coronary artery smooth muscle cell fibronectin protein synthesis and mRNA compared to host. By using neutralizing antibodies to IL-1 beta, fibronectin protein synthesis and mRNA levels were reduced in donor cells to the levels observed in the host cells and a similar reduction in synthesis was observed with the IL-1 receptor antagonist. Immunoprecipitation of newly synthesized IL-1 beta revealed increased endogenous levels in donor compared to host cells. We therefore suggest in the coronary arteriopathy a pathophysiologic mechanism whereby IL-1 beta-mediated increased fibronectin synthesis may promote lymphocyte trapping and migration of medial smooth muscle cells leading to progressive intimal thickening associated with the post-cardiac transplant coronary arteriopathy. PMID- 8408638 TI - Arginine feeding modifies cyclosporine nephrotoxicity in rats. AB - Glycine (G) infusion causes renal vasodilation mediated by nitric oxide (NO). Cyclosporine A (CsA) nephrotoxicity is characterized by preglomerular vasoconstriction and decreased efferent arteriolar tone probably related to reduced NO and angiotensin II, respectively. L-Arginine (ARG) is a precursor to NO. To test the hypothesis that chronic CsA decreases renal NO activity, we compared the glomerular hemodynamic response to glycine infusion in rats after 8 d of CsA (30 mg/kg per d s.c.), CsA and ARG (1.6 g/kg per d p.o.) (A/CsA), and in two groups of pair-fed controls (CON, A/CON). Single nephron GFR (SNGFR), single nephron plasma flow (SNPF), glomerular capillary hydrostatic pressure gradient (delta P), proximal tubular reabsorption (APR), and kidney tissue angiotensin II (AIIk) were measured before and during G. CsA was associated with baseline decrements in SNGFR, SNPF, delta P, and AIIk, and with a blunted hemodynamic response to G. In CON, ARG did not affect baseline hemodynamics or modify the response to G. In CsA, ARG decreased baseline preglomerular resistance and restored the glomerular hemodynamic response to G. G was associated with a significant increase in AIIk in both CON and CsA. These findings suggest that (a) CsA is associated with decreased AIIk, and (b) CsA may diminish NO activity within the kidney, and that this capacity may be partially restored by arginine feeding. PMID- 8408639 TI - Cellular heterogeneity of ammonium ion transport across the basolateral membrane of the hamster medullary thick ascending limb of Henle's loop. AB - The epithelia of the medullary thick ascending limb (MAL) consists of two cell types, high (HBC) and low basolateral conductance (LBC) cell, depending on the K+ conductance of the basolateral membrane. The NH4+ conductance distinct from the K+ conductance has been suggested to exist in the proximal tubule, MAL cell, and Xenopus oocyte. The present study was designed to examine whether there is a conductive NH4+ transport system distinct from K+ conductance in two different cell types of the hamster MAL perfused in vitro. The basolateral membrane voltage (VB) was measured by impaling cells with conventional microelectrodes. Addition of NH4+ to the bath depolarized VB in a dose-dependent manner in both cell types. The response was maintained in the absence of HCO3-. When the VB deflection elicited upon 50 mM KCl or NH4Cl in the bath (delta VBK+ or delta VBNH4+) were compared, delta VBNH4+ was almost the same as delta VBK+ in the HBC cell, whereas the former was greater than the latter in the LBC. In the HBC cell, 10 mM Ba2+ in the bath equally suppressed both delta VBK+ and delta VBNH4+, whereas in the LBC cell it suppressed delta VBK+ with a small effect on delta VBNH4+, indicating that NH4+ is transported via a pathway distinct from Ba(2+)-sensitive K+ conductance. The VB deflection by NH4+ was unaffected by addition of 0.1 mM ouabain or 10 microM 5-nitro-2-(3-phenylpropylamino)-benzoate (a Cl- channel blocker) to the bath, excluding the contribution of the Na+, K+ pump or Cl- channel. An abrupt reduction of Na+ in the bath from 200 to 20 mM did not cause any changes in VB, suggesting that a nonselective cation channel may not account for the NH4+ transport. Amiloride at 10 microM inhibited delta VBNH4+ with a higher efficacy in the LBC cell. We conclude that a rheogenic NH4+ transport system independent from the K+ conductance exists in the basolateral membrane of the LBC cell of the hamster MAL, and may play some roles in the regulation of NH4+ transport. PMID- 8408640 TI - Regulation of Na,K-adenosine triphosphatase gene expression by sodium ions in cultured neonatal rat cardiocytes. AB - Na,K-ATPase (Na,K-pump) plays an important role in the regulation of intracellular ion composition. The purpose of this study is to determine whether Na+ regulates the levels of mRNA coding for Na,K-ATPase alpha and beta subunits in cultured neonatal rat cardiocytes. We measured intracellular Na+ levels ([Na+]i) in cardiocytes using a Na(+)-sensitive fluorescence dye (SBFI). 1 mM ouabain caused a significant increase in [Na+]i in cardiocytes; from 12.8 +/- 0.3 to 28.8 +/- 1.8 mM. Exposure of cardiocytes to 1 mM ouabain resulted in a three- to fourfold increase in alpha 1, alpha 2, and alpha 3 mRNA accumulation, and an approximate two-fold increase in beta 1 mRNA accumulation. A maximum elevation was reached at 60 min in both cases. The ouabain-induced alpha 1 mRNA accumulation was still observed in the Ca(2+)-free culture medium. Exposure of cardiocytes to 10 microM monensin in the absence of extracellular Ca2+ also resulted in a threefold increase in alpha 1 mRNA accumulation. The increased alpha 1 mRNA expression by 1 mM ouabain was associated with a fourfold increase in alpha 1 subunit protein accumulation. Transfection experiments with chimeric plasmids containing 5'-flanking sequences of alpha 1, alpha 2, and alpha 3 isoform genes and a luciferase reporter gene revealed that 1 mM ouabain caused a twofold increase in luciferase activity in each alpha system. These results suggest that Na+ directly regulates Na,K-ATPase gene expression in cardiocytes. The transfection study further supports the premise that Na(+)-responsive elements are located within the 5'-flanking sequences of each alpha isoform gene. PMID- 8408641 TI - Evidence of direct estrogenic regulation of human corticotropin-releasing hormone gene expression. Potential implications for the sexual dimophism of the stress response and immune/inflammatory reaction. AB - Corticotropin-releasing hormone (CRH) plays major roles in coordination of the stress response and regulation of the immune/inflammatory reaction, two important functions associated with sexual dimorphism. Two overlapping segments of the 5' flanking region of the human (h) CRH gene, the proximal 0.9 kb (containing two perfect half-palindromic estrogen-responsive elements [EREs]) and the 2.4 kb (including the former and containing two additional perfect half-palindromic EREs), were examined for their ability to confer estrogen-mediated transcriptional enhancement to a homologous or heterologous promoter. The level of estrogen-induced transactivation by the 0.9- and 2.4-kb segments was determined by chloramphenicol acetyltransferase analysis in CV-1 cells cotransfected with estrogen receptor (ER) cDNA expression plasmids, and found to be respectively approximately 10% and 20% of that of the strongly estrogen responsive Xenopus vitellogenin A2 enhancer. Gel retardation and immunoprecipitation demonstrated specific association between the perfect half palindromic EREs of hCRH gene and the DNA binding domain of hER in vitro. These findings may constitute the basis of sexual dimorphism in the expression of the CRH gene in the central nervous system and periphery, and might shed light in existing gender differences in stress response and immune regulation. PMID- 8408642 TI - Comparison of the effects of recombinant human insulin-like growth factor-I and insulin on glucose and leucine kinetics in humans. AB - To compare the metabolic effects of elevated plasma concentrations of IGF-I and insulin, overnight-fasted normal subjects were studied twice, once receiving IGF I and once insulin at doses that resulted in identical increases in glucose uptake during 8-h euglycemic clamping. Recombinant human IGF-I or insulin were infused in one group at high doses (30 micrograms/kg per h IGF-I or 0.23 nmol/kg per h insulin) and in another group at low doses (5 micrograms/kg per h IGF-I or 0.04 nmol/kg per h insulin). Glucose rate of disappearance (measured by [6,6-D2] glucose infusions) increased from baseline by 239 +/- 16% during high dose IGF-I vs 197 +/- 18% during insulin (P = 0.021 vs IGF-I). Hepatic glucose production decreased by 37 +/- 6% during high dose IGF-I vs 89 +/- 13% during insulin (P = 0.0028 vs IGF-I). IGF-I suppressed whole body leucine flux ([1-13C]-leucine infusion technique) more than insulin (42 +/- 4 vs 32 +/- 3% during high doses, P = 0.0082). Leucine oxidation rate decreased during high dose IGF-I more than during insulin (55 +/- 4 vs 32 +/- 6%, P = 0.0001). The decreases of plasma concentrations of free fatty acids, acetoacetate, and beta-hydroxybutyrate after 8 h of IGF-I and insulin administration were similar. Plasma C-peptide levels decreased by 57 +/- 4% during high doses of IGF-I vs 36 +/- 6% during insulin (P = 0.005 vs IGF-I). The present data demonstrate that, compared to insulin, an acute increase in plasma IGF-I levels results in preferential enhancement of peripheral glucose utilization, diminished suppression of hepatic glucose production, augmented decrease of whole body protein breakdown (leucine flux), and of irreversible leucine catabolism but in similar antilipolytic effects. The data suggest that insulin-like effects of IGF-I in humans are mediated in part via IGF-I receptors and in part via insulin receptors. PMID- 8408643 TI - Enhanced stimulus-secretion coupling in polyamine-depleted rat insulinoma cells. An effect involving increased cytoplasmic Ca2+, inositol phosphate generation, and phorbol ester sensitivity. AB - To extend previous observations on the role of polyamines in insulin production, metabolism, and replication of insulin-secreting pancreatic beta cells, we have studied the role of polyamines in the regulation of the stimulus-secretion coupling of clonal rat insulinoma cells (RINm5F). For this purpose, RINm5F cells were partially depleted in their polyamine contents by use of the specific ornithine decarboxylase inhibitor difluoromethylornithine (DFMO), which led to an increase in cellular insulin and ATP contents. Analysis of different parts of the signal transduction pathway revealed that insulin secretion and the increase in cytoplasmic free Ca2+ concentration ([Ca2+]i) after K(+)-induced depolarization were markedly enhanced in DFMO-treated cells. These effects were paralleled by increased voltage-activated Ca2+ currents, as judged by whole-cell patch-clamp analysis, probably reflecting increased channel activity rather than elevated number of channels per cell. DFMO treatment also rendered phospholipase C in these cells more sensitive to the muscarinic receptor agonist carbamylcholine, as evidenced by enhanced generation of inositol phosphates, increase in [Ca2+]i and insulin secretion, despite an unaltered ligand binding to muscarinic receptors and lack of effect on protein kinase C activity. In addition, the tumor promoter 12-O-tetradecanoylphorbol 13-acetate, at concentrations suggested to be specific for protein kinase C activation, evoked an increased insulin output in polyamine deprived cells compared to control cells. The stimulatory effects of glucose or the cyclic AMP raising agent theophylline on insulin release were not increased by DFMO treatment. In spite of increased binding of sulfonylurea in DFMO-treated cells, there was no secretory response or altered increase in [Ca2+]i in response to the drug in these cells. It is concluded that partial polyamine depletion sensitizes the stimulus-secretion coupling at multiple levels in the insulinoma cells, including increased voltage-dependent Ca2+ influx and enhanced responsiveness to activators of phospholipase C and protein kinase C. In their entirety, our present results indicate that the behavior of the stimulus secretion coupling of polyamine-depleted RINm5F insulinoma cells changes towards that of native beta cells, thus improving the usefulness of this cell line for studies of beta cell insulin secretion. PMID- 8408644 TI - Antitumor effects of interleukin-7 and adoptive immunotherapy on human colon carcinoma xenografts. AB - The antitumor properties of recombinant human IL-7 (rhIL-7) on a human tumor was evaluated by engrafting a human colon carcinoma into immunodeficient mice and then treating the mice with rhIL-7 and adoptively transferred human peripheral blood T cells. It was found that rhIL-7 alone had no effect on the survival of the tumor-bearing recipients. However, the combination of rhIL-7 and human T cells significantly promoted the survival of the recipients compared with mice receiving either treatment by itself. When the surviving mice were analyzed 6 mo later for the degree of human cell engraftment, the recipients receiving both rhIL-7 and human T cells had greater numbers of human CD8+ T cells in the spleens. However, the human T cells recovered from the surviving mice showed low lytic activity against the tumor in vitro. Supernatants from human T cells cultured with the tumor and rhIL-7 in vitro were found to inhibit tumor growth and were demonstrated to contain high levels of IFN-gamma. Antibodies to IFN gamma neutralized the growth inhibition of the tumor both in vitro and in vivo demonstrating that the in vivo mechanism underlying the antitumor effects of this regimen was partly dependent on the production of IFN-gamma by the T cells and not their cytolytic capability. Interestingly, systemic administration of rhIFN gamma to tumor-bearing mice yielded little antitumor effect suggesting that adoptive immunotherapy with rhIL-7 was superior possibly because of the continuous local release of the cytokines. Therefore, rhIL-7 may be of clinical use as an antineoplastic agent and the human/mouse model is a potentially important preclinical model for in vivo evaluation of the efficacy of this and other immunotherapies. PMID- 8408645 TI - Subcellular localization of Bcr, Abl, and Bcr-Abl proteins in normal and leukemic cells and correlation of expression with myeloid differentiation. AB - We used specific antisera and immunohistochemical methods to investigate the subcellular localization and expression of Bcr, Abl, and Bcr-Abl proteins in leukemic cell lines and in fresh human leukemic and normal samples at various stages of myeloid differentiation. Earlier studies of the subcellular localization of transfected murine type IV c-Abl protein in fibroblasts have shown that this molecule resides largely in the nucleus, whereas transforming deletion variants are localized exclusively in the cytoplasm. Here, we demonstrate that the murine type IV c-Abl protein is also found in the nucleus when overexpressed in a mouse hematopoietic cell line. However, in both normal and leukemic human hematopoietic cells, c-Abl is discerned predominantly in the cytoplasm, with nuclear staining present, albeit at a lower level. In contrast, normal endogenous Bcr protein, as well as the aberrant p210BCR-ABL and p190BCR ABL proteins consistently localize to the cytoplasm in both cell lines and fresh cells. The results with p210BCR-ABL were confirmed in a unique Ph1-positive chronic myelogenous leukemia (CML) cell line, KBM5, which lacks the normal chromosome 9 and hence the normal c-Abl product. Because the p210BCR-ABL protein appears cytoplasmic in both chronic phase and blast crisis CML cells, as does the p190BCR-ABL in Ph1-positive acute leukemia, a change in subcellular location of Bcr-Abl proteins between cytoplasm and nucleus cannot explain the different spectrum of leukemias associated with p210 and p190, nor the transition from the chronic to the acute leukemia phenotype seen in CML. Further analysis of fresh CML and normal hematopoietic bone marrow cells reveals that p210BCR-ABL, as well as the normal Bcr and Abl proteins, are expressed primarily in the early stages of myeloid maturation, and that levels of expression are reduced significantly as the cells mature to polymorphonuclear leukocytes. Similarly, a decrease in Bcr and Abl levels occurs in HL-60 cells induced by DMSO to undergo granulocytic differentiation. The action of p210BCR-ABL and its normal counterparts may, therefore, take place during the earlier stages of myeloid development. PMID- 8408646 TI - Exogenous adenosine triphosphate (ATP) preserves proximal tubule microfilament structure and function in vivo in a maleic acid model of ATP depletion. AB - The hallmark of ischemic acute renal failure is a rapid and early decline in proximal tubule ATP. Since we have previously shown that over half of apical microfilament losses occur within the first 5 min of experimental ischemic injury, we postulated that microfilament (F-actin) structure and cellular location are dependent on cellular ATP levels. To test this hypothesis, we used maleic acid to selectively inhibit renal cortical ATP production in vivo. Maleic acid significantly decreased tissue ATP and apical F-actin in a dose-dependent manner relative to equimolar sodium chloride controls, yet higher doses of maleic acid quantitatively resulted in net actin polymerization, primarily in the cytoplasm. Functionally, maleic acid decreased glomerular filtration rate (GFR) and tubular reabsorption of sodium (TRNa) in a dose-dependent manner relative to sodium chloride controls. Administration of exogenous ATP resulted in significant increases in tissue ATP, net actin depolymerization, and relocation of F-actin from the cytoplasm back to the apical surface coinciding with increases in GFR and TRNa. Thus, ATP depletion induced by maleic acid resulted in significant cytoskeletal and functional alterations that were ameliorated by exogenous ATP. We therefore conclude that the structure and cellular location of F-actin necessary for normal functioning of proximal tubule cells in vivo is dependent on tissue ATP levels. PMID- 8408647 TI - GLUT-2 function in glucose-unresponsive beta cells of dexamethasone-induced diabetes in rats. AB - Spontaneous and dexamethasone-induced noninsulin-dependent diabetes mellitus (NIDDM) in rats is associated with loss of glucose-stimulated insulin secretion (GSIS) and a reduction in both GLUT-2-positive beta cells and high Km glucose transport. To determine if the chronology and correlation of these abnormalities is consistent with a causal relationship, Zucker (fa/fa) rats were studied longitudinally before and during 10 d of dexamethasone-induced (0.4 mg/kg per d i.p.) NIDDM. Within 24 h of dexamethasone treatment blood glucose rose and GSIS declined, becoming paradoxically negative (-87 +/- 12 microU/ml per min) on day 10. Blood glucose was negatively correlated with GSIS (r = -0.92; P < 0.001). 3-0 methyl-D-glucose (3MG) transport was unchanged at 12 h, 23% below normal on day 1, and declined further to a nadir 59% below normal. The GLUT-2-positive beta cell area did not decline until 48 h, reaching a nadir of 35% of normal at 10 d. The area of GLUT-2-positive beta cells was correlated with GSIS (r = 0.77; P < 0.005). We conclude that the chronology and correlation between GSIS loss and hyperglycemia is consistent with a cause-effect relationship, but that the subtotal impairment in glucose transport by itself cannot explain the total loss of GSIS if one assumes that normal beta cells are functionally homogenous. PMID- 8408648 TI - Infection of human synovial cells by human T cell lymphotropic virus type I. Proliferation and granulocyte/macrophage colony-stimulating factor production by synovial cells. AB - The present study was performed to clarify the relationship between human T cell lymphotropic virus type I (HTLV-I) infection and chronic inflammatory arthropathy. To determine the ability of HTLV-I to infect synovial cells and the effect on synovial cell proliferation, synovial cells were cocultured with the HTLV-I-producing T cell lines (MT-2 or HCT-1). After coculture with HTLV-I infected T cells, the synovial cells expressed HTLV-I-specific core antigens, and HTLV-I proviral DNA was detected from the synovial cells by polymerase chain reaction. These cocultured synovial cells with HTLV-I-infected T cells proliferated more actively than the synovial cells cocultured with uninfected T cells. This stimulatory effect of HTLV-I-infected T cells on synovial cell proliferation seems necessary to contact each other. After being cocultured with MT-2 cells, synovial cells proliferated more actively than control cells even after several passages. Furthermore, HTLV-I-infected synovial cells produced significant amounts of granulocyte/macrophage colony-stimulating factor. These results suggest that HTLV-I can infect synovial cells, resulting their active proliferation and may be involved in the pathogenesis of proliferative synovitis similar to that found in rheumatoid arthritis. PMID- 8408649 TI - In the absence of other Fc receptors, Fc gamma RIIIA transmits a phagocytic signal that requires the cytoplasmic domain of its gamma subunit. AB - The transmembrane isoform of Fc gamma RIII, Fc gamma RIIIA, is found on NK cells, cultured monocytes, and tissue macrophages in association with a dimer of an accessory subunit, either gamma or zeta. Functions of individual Fc receptors have been difficult to analyze due to coexpression of the receptors on hematopoietic cells and permanent cell lines expressing Fc receptors. cDNAs for the alpha and gamma subunits of Fc gamma RIIIA were cotransfected into COS-1 cells, which lack endogenous Fc receptors, to evaluate receptor-mediated phagocytosis and changes in [Ca2+]i. Transfectants both bound and phagocytosed IgG-sensitized erythrocytes and, following activation of Fc gamma RIIIA, increased [Ca2+]i. The gamma subunit was essential both for the surface expression of the receptor and for transduction of the phagocytic signal. Truncation of the gamma subunit cytoplasmic domain (amino acids 65-80) eliminated phagocytic function. Phorbol ester inhibited phagocytosis in a concentration dependent manner, but did not affect IgG-sensitized erythrocytes binding, suggesting that a protein kinase C-dependent pathway inhibits phagocytosis. The data indicate that a tyrosine containing cytoplasmic domain within the gamma subunit is required for phagocytosis by Fc gamma RIIIA. PMID- 8408650 TI - Glucocorticoid deficiency increases phospholipase A2 activity in rats. AB - An important mechanism for the antiinflammatory effect of pharmacological doses of glucocorticoids is the inhibition of arachidonic acid release from phospholipids by phospholipase A2 (PLA2). As a corollary, one might predict that low endogenous concentrations of glucocorticoids favor inflammatory disease states. Indeed, clinical and experimental observations revealed an association between glucocorticoid deficiency and disease states caused by immunological and/or inflammatory mechanisms. The purpose of the present investigation was to study the regulation of PLA2 mRNA, protein, and enzyme activity in adrenalectomized (ADX) rats where glucocorticoid concentrations were below physiological levels. The mRNA of group I and II PLA2 were measured by PCR. Group II PLA2 mRNA was increased by 126 +/- 9% in lung tissue of ADX rats, whereas group I PLA2 was increased only by 27 +/- 1.5%. The increase in group II mRNA in ADX rats was reflected by a corresponding increase of group II PLA2 protein (70 100%) in lung, spleen, liver, and kidney. This increase was reversed by the administration of exogenous corticosterone. After ADX, the percentage increase in total PLA2 activity was higher than that of mRNA or PLA2 protein, suggesting that the activity of the enzyme was modulated by inhibitors or activators. The concentration of lipocortin-I, an inhibitor of PLA2 enzyme was strongly correlated with the activity of PLA2 in the tissues (lung, spleen, liver, and kidney). In all these tissues, the concentrations of lipocortin-I declined after ADX. Thus upregulation of PLA2 enzyme and downregulation of lipocortin-I might account for the enhanced inflammatory response in hypoglucocorticoid states. PMID- 8408651 TI - Disparate effects of insulin on isolated rabbit afferent and efferent arterioles. AB - Despite evidence that insulin per se may be an important regulator of glomerular hemodynamics, little is known about its direct action on the glomerular afferent arterioles (Af-Art) and efferent arterioles (Ef-Art), the crucial vascular segments that control glomerular hemodynamics. In the present study, we examined the direct effect of physiological concentrations of insulin on isolated microperfused rabbit Af- and Ef-Arts. After cannulation, vessels were equilibrated in insulin-free medium for 30 min. To determine whether insulin causes vasodilation or constriction, increasing doses (5, 20, and 200 microU/ml) were added to the bath and lumen of arterioles that were either preconstricted to 50% of control diameter with norepinephrine or left nonpreconstricted. Insulin caused no vasoconstriction in either Af- or Ef-Arts, but it reversed norepinephrine-induced constriction in Ef-Arts but not Af-Arts (suggesting a vasodilator action selective to the Ef-Art): at 200 microU/ml, insulin increased Ef-Art luminal diameter by 75.8 +/- 7.0% from the preconstricted level (n = 6; P < 0.008). The vasorelaxant effect of insulin on Ef-Arts was not affected by blockade of either endothelium-derived relaxing factor/nitric oxide or prostaglandin synthesis. Despite the lack of effect of insulin on Af-Art when added after the equilibration period, when Af-Arts were equilibrated in the presence of either 20 or 200 microU/ml insulin, their basal diameter was significantly reduced (11.7 +/- 0.9 microns; P < 0.025, n = 6, and 12.0 +/- 0.9 microns; P < 0.025, n = 7, respectively) compared with nontreated Af-Arts (16.2 +/- 1.3 microns; n = 7). In conclusion, this study demonstrates that at physiological concentrations, insulin dilates NE-constricted Ef-Arts, while insulin pretreatment enhances Af-Art tone. The disparate actions of insulin on the Af- vs the Ef-Art may contribute to its beneficial effect on glomerular hypertension. PMID- 8408652 TI - Interaction of human beta 1 thyroid hormone receptor and its mutants with DNA and retinoid X receptor beta. T3 response element-dependent dominant negative potency. AB - Mutations in the human beta thyroid hormone receptor (h-TR beta) gene are associated with the syndrome of generalized resistance to thyroid hormone. We investigated the interaction of three h-TR beta 1 mutants representing different types of functional impairment (kindreds ED, OK, and PV) with different response elements for 3,3',5-triiodothyronine (T3) and with retinoid X receptor beta (RXR beta). The mutant receptors showed an increased tendency to form homodimers on a palindromic T3-response element (TREpal), a direct repeat (DR + 4), and an inverted palindrome (TRElap). On TRElap, wild type TR binding was decreased by T3, while the mutant receptors showed a variably decreased degree of dissociation from TRElap in response to T3. The extent of dissociation was proportional to their T3 binding affinities. RXR beta induced the formation of h-TR beta 1:RXR beta heterodimers equally well for mutants and the wild type h-TR beta 1 on these T3 response elements. However, the T3-dependent increase in heterodimerization with RXR beta was absent or reduced for the mutant TRs. Transient transfection studies indicated that the dominant negative potency was several-fold more pronounced on the TRElap as compared to TREpal or DR + 4. In CV-1 and HeLa cells, transfection of RXR beta could not reverse the dominant negative action. These results demonstrate that the binding of mutant h-TRs to DNA, as well as their dominant negative potency, are TRE dependent. In addition, competition for DNA binding, rather than for limiting amounts of RXR beta, is likely to mediate the dominant negative action. PMID- 8408653 TI - Defective splicing of mRNA from one COL1A1 allele of type I collagen in nondeforming (type I) osteogenesis imperfecta. AB - Osteogenesis imperfecta (OI) type I is the mildest form of heritable bone fragility resulting from mutations within the COL1A1 gene. We studied fibroblasts established from a child with OI type I and demonstrated underproduction of alpha 1 (I) collagen chains and alpha 1 (I) mRNA. Indirect RNase protection suggested two species of alpha 1 (I) mRNA, one of which was not collinear with fully spliced alpha 1 (I) mRNA. The noncollinear population was confined to the nuclear compartment of the cell, and contained the entire sequence of intron 26 and a G- >A transition in the first position of the intron donor site. The G-->A transition was also identified in the genomic DNA. The retained intron contained an in-frame stop codon and introduced an out-of-frame insertion within the collagen mRNA producing stop codons downstream of the insertion. These changes probably account for the failure of the mutant RNA to appear in the cytoplasm. Unlike other splice site mutations within collagen mRNA that resulted in exon skipping and a truncated but inframe RNA transcript, this mutation did not result in production of a defective collagen pro alpha 1 (I) chain. Instead, the mild nature of the disease in this case reflects failure to process the defective mRNA and thus the absence of a protein product from the mutant allele. PMID- 8408654 TI - Antithrombotic effects of thrombin-induced activation of endogenous protein C in primates. AB - The effects on thrombosis and hemostasis of thrombin-induced activation of endogenous protein C (PC) were evaluated in baboons. Thrombosis was induced by placing into arteriovenous shunts a segment of Dacron vascular graft, which generated arterial platelet-rich thrombus, followed by an expansion region of low shear blood flow, which in turn accumulated fibrin-rich venous-type thrombus. Thrombosis was quantified by 111In-platelet imaging and 125I-fibrinogen accumulation. Intravenous infusion of alpha-thrombin, 1-2 U/kg-min for 1 h, increased baseline activated PC levels (approximately 5 ng/ml) to 250-500 ng/ml (P < 0.01). The lower thrombin dose, which did not deplete circulating platelets, fibrinogen, or PC, reduced arterial graft platelet deposition by 48% (P < 0.05), and platelet and fibrin incorporation into venous-type thrombus by > 85% (P < 0.01). Thrombin infusion prolonged the activated partial thromboplastin clotting time, elevated fibrinopeptide A (FPA), thrombin-antithrombin III complex (T:AT III), and fibrin D-dimer plasma levels (P < 0.01), but did not affect bleeding times. Thrombin's antithrombotic effects were blocked by infusing a monoclonal antibody (HPC-4) which prevented PC activation in vivo, caused shunt occlusion, increased the consumption of platelets and fibrinogen, elevated plasma FPA and T:AT III levels, and reduced factor VIII (but not factor V) procoagulant activity (P < 0.05). We conclude that activated PC is a physiologic inhibitor of thrombosis, and that activation of endogenous PC may represent a novel and effective antithrombotic strategy. PMID- 8408655 TI - Fluid shear stress differentially modulates expression of genes encoding basic fibroblast growth factor and platelet-derived growth factor B chain in vascular endothelium. AB - Fluid shear stress has been shown to be an important regulator of vascular structure and function through its effect on the endothelial cell. We have explored the effect of shear stress on the expression of the heparin-binding growth factors platelet-derived growth factor B chain (PDGF-B) and basic fibroblast growth factor (bFGF) in bovine aortic endothelial cells using a purpose-built cone-plate viscometer. Using morphometric analysis, we have mimicked the endothelial cell shape changes encountered in vivo in response to shear stress and correlated these with changes in gene expression. Steady laminar shear stress of 15 and 36 dyn/cm2 both resulted in endothelial cell shape change, but the higher shear stress induced greater and more uniform alignment in the direction of flow and nuclear protrusion after 24 h. Steady laminar shear stress of both 15 and 36 dyn/cm2 induced a significant 3.9- and 4.2-fold decrease, respectively, in PDGF-B mRNA at 9 h. In contrast, steady laminar shear of 15 dyn/cm2 induced a mild and transient 1.5-fold increase in bFGF mRNA while shear of 36 dyn/cm2 induced a significant 4.8-fold increase at 6 h of shear which remained at 2.9-fold at 9 h. Pulsatile and turbulent shear stress showed the same effect as steady laminar shear stress (all at 15 dyn/cm2 time-average magnitude) on PDGF-B and bFGF mRNA content. Cyclic stretch (20% strain, 20/min) of cells grown on silicone substrate did not significantly affect either PDGF-B or bFGF mRNA levels. These results suggest that expression of each peptide growth factor gene is differentially regulated by fluid shear stress in the vascular endothelial cell. These results may have implications on vascular structure and function in response to hemodynamic forces and present a model for the study of transduction of mechanical stimuli into altered gene expression. PMID- 8408656 TI - Cell-specific expression of alpha 1-antitrypsin in human intestinal epithelium. AB - alpha 1-Antitrypsin (alpha 1-AT) is an acute phase plasma protein predominantly derived from the liver which inhibits neutrophil elastase. Previous studies have suggested that alpha 1-AT is also expressed in human enterocytes because alpha 1 AT mRNA could be detected in human jejunum by RNA blot analysis, and alpha 1-AT synthesis could be detected in a human intestinal adenocarcinoma cell line Caco2, which spontaneously differentiates into villous-like enterocytes in tissue culture. To definitively determine that the alpha 1-AT gene is expressed in human enterocytes in vivo, we examined tissue slices of human jejunum and ileum by in situ hybridization. The results demonstrate specific hybridization to enterocytes from the bases to the tips of the villi. Although there was no hybridization to enterocytes in most of the crypt epithelium, there was intense specific hybridization in one region of the crypt. Double-label immunohistochemical studies showed that alpha 1-AT and lysozyme co-localized to this region, indicating that it represented Paneth cells. Finally, there was a marked increase in hybridization to alpha 1-AT mRNA in villous enterocytes and Paneth cells in Crohn's disease. The results of this study provide definitive evidence that alpha 1-AT is expressed in human jejunal and ileal enterocytes in vivo, and show that alpha 1-AT is also a product of Paneth cells. Together with the results of other studies, these data raise the possibility that alpha 1-AT detected in fecal alpha 1-AT clearance assays for diagnosing protein-losing enteropathies is predominantly derived from sloughed enterocytes. PMID- 8408657 TI - Concurrent expression of erythroid and renal aquaporin CHIP and appearance of water channel activity in perinatal rats. AB - Major phenotypic changes occur in red cell membranes during the perinatal period, but the underlying molecular explanations remain poorly defined. Aquaporin CHIP, the major erythroid and renal water channel, was studied in perinatal rats using affinity-purified anti-CHIP IgG for immunoblotting, flow cytometry, and immunofluorescence microscopy. CHIP was not detected in prenatal red cells but was first identified in circulating red cells on the third postnatal day. Most circulating red cells were positive for CHIP by the seventh postnatal day, and this proportion rose to nearly 100% by the 14th day. The ontogeny of red cell CHIP correlated directly with acquisition of osmotic water permeability and inversely with Arrhenius activation energy. Only minor alterations in the composition of red cell membrane lipids occurred at this time. Immunohistochemical analysis of perinatal kidneys demonstrated a major induction of CHIP in renal proximal tubules and descending thin limbs at birth, coincident with the development of renal concentration mechanisms. Therefore, water channels are unnecessary for oxygen delivery or survival in the prenatal circulation, however CHIP may confer red cells with the ability to rehydrate rapidly after traversing the renal medulla, which becomes hypertonic after birth. PMID- 8408658 TI - Vasonatrin peptide: a unique synthetic natriuretic and vasorelaxing peptide. AB - This study reports the cardiovascular and renal actions of a novel and newly synthesized 27-amino acid peptide termed vasonatrin peptide (VNP). VNP is a chimera of atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP). This synthetic peptide possesses the 22-amino acid structure of CNP, which is a cardiovascular selective peptide of endothelial origin and is structurally related to ANP. VNP also possesses the five-amino acid COOH terminus of ANP. The current study demonstrates both in vitro and in vivo that VNP possesses the venodilating actions of CNP, the natriuretic actions of ANP, and unique arterial vasodilating actions not associated with either ANP or CNP. PMID- 8408659 TI - A missense mutation in the cholesteryl ester transfer protein gene with possible dominant effects on plasma high density lipoproteins. AB - Plasma HDL are a negative risk factor for atherosclerosis. Cholesteryl ester transfer protein (CETP; 476 amino acids) transfers cholesteryl ester from HDL to other lipoproteins. Subjects with homozygous CETP deficiency caused by a gene splicing defect have markedly elevated HDL; however, heterozygotes have only mild increases in HDL. We describe two probands with a CETP missense mutation (442 D:G). Although heterozygous, they have threefold increases in HDL concentration and markedly decreased plasma CETP mass and activity, suggesting that the mutation has dominant effects on CETP and HDL in vivo. Cellular expression of mutant cDNA results in secretion of only 30% of wild type CETP activity. Moreover, coexpression of wild type and mutant cDNAs leads to inhibition of wild type secretion and activity. The dominant effects of the CETP missense mutation during cellular expression probably explains why the probands have markedly increased HDL in the heterozygous state, and suggests that the active molecular species of CETP may be multimeric. PMID- 8408660 TI - Conserved cytoplasmic tyrosine residues of the gamma subunit are required for a phagocytic signal mediated by Fc gamma RIIIA. AB - Fc receptors for immunoglobulins are found on many cells and are important in host defense. We transfected Fc gamma RIIIA, present on macrophages and natural killer (NK) cells, into COS-1 cells to study its role in phagocytosis and calcium mobilization in the absence of other Fc gamma receptors. Human Fc gamma RIIIA alpha (CD16) was cotransfected with its associated chains, either Fc gamma RIIIA gamma or zeta. Both gamma and zeta were observed to induce a phagocytic signal, but gamma was at least sixfold more effective than zeta. Conservative substitution by phenylalanine of either one of the two cytoplasmic tyrosine residues in the gamma chain resulted in markedly diminished phagocytosis and calcium mobilization. Tyrphostin 23, an inhibitor of tyrosine kinases, reversibly inhibited phagocytosis. Further, in vitro kinase assays with the wild type and mutant gamma chains demonstrated that the wild type gamma chain, but not the mutant gamma chains, is phosphorylated. These results suggest that the cytoplasmic tyrosine residues and tyrosine phosphorylation are required for Fc gamma RIIIA to mediate two signal transduction events: phagocytosis and calcium mobilization. PMID- 8408661 TI - Heparin regulates endothelin production through endothelium-derived nitric oxide in human endothelial cells. AB - Heparin shows blood pressure lowering effect in hypertensive patients and animal models. The present study examined the effect of heparin on vasoconstrictor endothelin-1 (ET-1) production in cultured human umbilical vein endothelial cells (ECs) to elucidate the mechanism of antihypertensive effect of heparin. Heparin suppressed both basal and thrombin-stimulated ET-1 mRNA expression paralleled with a decrease in ET-1 peptide release in a dose-dependent manner. Heparin concomitantly enhanced nitric oxide (NO) formation measured by NO2/NO3 levels and cGMP production in ECs. These enhancements were more marked when ECs were stimulated by thrombin. However, these heparin's effects were blunted in the presence of endothelium-derived nitric oxide (EDNO) synthesizing inhibitor NG monomethyl L-arginine. Therefore, these results suggest that suppression of ET-1 production by heparin is EDNO mediated. PMID- 8408662 TI - Clinical spectrum of anorexia nervosa in children. AB - A retrospective review of 21 patients ages 12 years and younger (age of onset range 7 to 12 years) with anorexia nervosa showed diagnostic delay in the youngest ones, high incidence of family psychiatric history, a remarkable severity of illness, and positive response to intensive treatment. Additional findings included significant comorbidity, a distinct subgroup with personality disorder and another with features of the "vulnerable child syndrome." This broad clinical spectrum of anorexia nervosa in children may explain the great variability in outcome. The development of anorexia nervosa in children relates to a complex combination of etiological and trigger factors. Precipitants identified in this study were physical maturation, entry into junior high, loss, or some combination thereof. PMID- 8408663 TI - Cognitive coping strategies of children with chronic illness. AB - Children with chronic illness need to adapt to more stresses than do healthy children. Research highlights the problems of children with chronic illnesses but not how they cope in response to the stress created by these problems. Cognitive appraisal of a stressor and of response options is an important aspect of coping. Our cross-sectional study investigated whether children with chronic illness used cognitive strategies for coping as often as did healthy children. One hundred seventy five children from summer camps with juvenile arthritis, asthma, or diabetes were compared with 145 healthy school children. Spontaneous responses to common painful and stressful events were categorized into coping or catastrophizing ideation. Data on anxiety, disease severity, and other sociodemographic variables were obtained. Coping strategies were reported by 64% of children with chronic illness and 63% of healthy children and varied significantly with age (p < .05) in both groups. Children with different chronic illnesses performed similarly except for a trend among children with severe juvenile rheumatoid arthritis who had higher rates of coping. For the individual stressful events, the rate of coping varied from 46% to 86%. The highest rates of coping responses were found with the child's recent personal stressful event where adolescents with chronic illness were twice as likely to offer more complex coping responses. More children with chronic illness than healthy children offered coping strategies in response to venipuncture (p < .001) but not to dental injection. Children with chronic illness report coping as their predominant strategy for adapting to common painful and stressful events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408664 TI - Child care centers: a community resource for injury prevention. AB - Child passenger safety, a major public health concern, has been addressed by state laws mandating use of child restraint devices. Usage rates in poor and minority communities are disproportionately low. To determine the influence of the child care center within these communities to improve routine use, an educational clinical trial, based on social learning theory was designed. Two urban child care centers enrolling high-risk 2- through 6-year-old children were monitored for correct child restraint use during a 5-month educational intervention at one center. Key features of intervention programming included weekly, developmentally appropriate presentations by the staff to the children and documented parental awareness of the child care center's policy advisory regarding child restraint use. Results demonstrate statistically significant (p < .01) increases in use at the intervention center. This study finds that child care center policy and programming can be effective in promoting child passenger safety. PMID- 8408665 TI - The Early Infancy Temperament Questionnaire. AB - Although there are several scales routinely used for assessment of temperament during the first year of life, none of them is well suited to the infant younger than 4 months old. The Early Infancy Temperament Questionnaire (EITQ) was designed to meet this need. The EITQ is a 76-item parent questionnaire for assessing the nine New York Longitudinal Study temperament characteristics in 1- to 4-month-old infants. The majority of the items were adapted from the Revised Infant Temperament Questionnaire to be developmentally appropriate for the very young infant. The standardization population consisted of 404 infants from one pediatric practice. Means for the nine categories were calculated separately for infants from 1 to 2 months and 3 to 4 months old. Internal consistency for the nine categories ranged from .42 to .76. Test-retest scores, completed between 2 to 3 weeks after the first rating, ranged from .43 to .87, with generally increasing retest levels in the older age group. None of the categories showed significant differences between male and female infants. This newly developed instrument should enhance the ability of both researchers and clinicians to assess temperament reliably and to understand better its contribution to clinical problems in the very young child. PMID- 8408666 TI - Chemotherapy induced nausea and emesis in pediatric cancer patients: external validity of child and parent emesis ratings. AB - Children's and parent's subjective ratings of the frequency and severity of nausea and emesis were assessed among 33 children with acute lymphoblastic leukemia receiving identical chemotherapy. Parents were trained to record the frequency of the child's actual emesis episodes during chemotherapy. Although parent and child ratings of nausea were significantly correlated, children generally rated their nausea and emesis as more frequent and more severe than did their parents. Parent ratings showed inadequate external validity when compared with behavioral observations. Children with greater anxiety and higher subjective ratings subsequently exhibited more frequent episodes of emesis by observation, suggesting that their perceptions of symptoms based on previous chemotherapy experiences may predict emesis during different chemotherapy. In a stepwise multiple regression analysis, antiemetic regimen and the child's anxiety as rated by the parent combined to account for approximately 47% of the variance in number of episodes of emesis. These findings are discussed in the context of factors limiting validity of parent and child reports of children's symptomatology with implications for future epidemiologic and intervention research. PMID- 8408667 TI - Cognitive development of preterm low birth weight children at 5 to 8 years old. AB - Prematurity and low birth weight have been considered to be important risk factors for cognitive development during early childhood; however, it has been suggested that the developmental delays disappear with age. Eighty-one preterm (< 38 weeks) low birth weight (< 2500 g) children between 5 and 8 years old from the Children of the National Longitudinal Survey of Youth were compared with individually matched full-term normal birth weight children to investigate the catch-up delays in cognitive functioning including language and mathematics skills. Preterm children showed a significant delay in cognitive functioning only until 6 years old. Regression analyses showed that environmental factors accounted for more variation in cognitive development than did perinatal factors. In support of a transactional model, preterm children exhibited a self-righting tendency during their early childhood so that eventually environmental influences overshadowed biological influences. PMID- 8408668 TI - Collecting data by telephone interviewing. PMID- 8408669 TI - Predictors of frequent middle school health room use. AB - Disproportionately high use of school health room (HR) services by small groups of users has been reported. This study investigated predictors of frequent HR use in two suburban middle schools involving 1089 students who made at least one HR visit during a single academic year. Subsets of high and low users were compared using logistic regression. Predictor variables included gender, grade, academic ability, and existence of a chronic health condition. Although demographic characteristics of the study schools varied significantly, a pattern of increasing HR use associated with progressive lowering of academic ability was demonstrated in both schools. Existence of a chronic health condition was associated with increased HR use, even when controlling for routine medication visits. Gender and grade were not predictive. These findings suggest that HR use reflects more than medical concerns. Patterns of HR use by students with chronic health conditions deserve additional study to determine whether current strategies to meet their needs are adequate. PMID- 8408670 TI - Normality: a clinically useless concept. The case of infant crying and colic. AB - To summarize, it has been argued that: 1. The assessment of complaints about crying and colic present particular diagnostic problems. 2. The crying brought as a complaint seldom indicates disease. 3. Once clinical disease has been ruled out, the clinical meanings of normality and abnormality no longer apply. 4. At that point, one should not try to determine a "cut-off" point for abnormal crying, because (a) it is unhelpful clinically, (b) it is wrong in principle, and (c) it is not likely that any specific amount of crying is normal or abnormal, independent of context. 5. As a possible alternative, it is proposed that we should think of the behavior not a symptom of something the infant "has," but as something the infant "does." This behavior may have consequences that are functional or dysfunctional for the infant, the caregiver, or the infant caregiver interaction. If this argument has merit, it may have some interesting and important implications for the way we think about, treat, and investigate developmental and behavioral problems including (but not limited to) infant crying and colic. First, what holds true for crying and colic may also hold for bedwetting and enuresis, overactivity and attention-deficit hyperactivity disorder, and abdominal pain and recurrent abdominal pain syndrome, to name just a few. As a brief test of their applicability, one might ask how often organic disease is found in these entities, or how often patients are investigated and treated because an arbitrary amount of these behaviors is taken to be "excessive" or abnormal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408671 TI - Psychosocial preventive intervention for families with parental mood disorder: strategies for the clinician. PMID- 8408672 TI - Population recovery capabilities of 35 cluster analysis methods. AB - Comparative evaluation of population recovery capabilities of 35 cluster analysis methods defined by different combinations of 5 profile similarity measures and 7 agglomeration rules was undertaken using artificial data that represented duplicate mixture samples from 4 latent populations. The latent population mean profiles differed primarily in elevation or in pattern parameters. Latent population sampling variances were controlled to provide two different levels of realistic overlap. The within-population distributions were multivariate normal with diagonal covariance structure. Across all conditions examined, complete linkage and Ward's minimum variance methods, used with Euclidian or city block interprofile distance measures, performed best. Single linkage, median, and centroid methods were substantially inferior for clustering individuals in accordance with true population memberships. PMID- 8408673 TI - Depression as a confounding variable in the estimation of habitual sleep time. AB - Self-reported habitual sleep time is an important variable because short and long sleep times are associated with mortality. Speculation with regard to these results usually focuses on physical health, rather than psychological factors. We investigated the role of anxiety and depression in reports of habitual sleep times by examining the relative and absolute discrepancy between individuals' initial estimates of their sleep times and sleep diaries made over a 2-week period. Results indicated that depressed mood was associated not only with a tendency initially to underestimate length of sleep (relative discrepancy), but also to exaggerate reported sleep time regardless of direction (absolute discrepancy). These results imply that studies that examine relationships between reported sleep times and mortality should take mental health factors into account. PMID- 8408674 TI - Personality, marital, and occupational factors associated with quality of life. AB - In three studies that employed community-based samples the relationship between personality, marital, and job-related factors and quality of life was examined. Study 1 indicated that hardiness and self-esteem were important components of overall quality of life. The marital communication skills of expressiveness and intimacy were identified as major aspects of overall quality of life in the second study. In Study 3, satisfaction with various job characteristics was related to overall quality of life. These studies start to provide definition to the quality of life concept in terms of personality characteristics, skills, and beliefs that have potential for modification. PMID- 8408675 TI - Comparison of DSM-II and DSM-III schizophrenics: a longitudinal perspective. AB - Patients who meet DSM-III criteria for schizophrenia were compared with those who meet DSM-II, but not DSM-III, criteria. Comparisons included measures of positive symptoms, negative symptoms, and social competence both before extended hospitalization and during a 7-year follow-up. The measure of positive symptoms was the only variable that differed significantly between groups at initial assessment. Positive symptoms were more stable and predictive for DSM-III schizophrenics than for DSM-II schizophrenics. The measure of negative symptoms was the most stable and predictive variable for the DSM-II schizophrenics. Increased homogeneity for the DSM-III schizophrenics was not found for most measures. PMID- 8408676 TI - Psychometric properties of the Separation-Individuation Test of Adolescence within a clinical population. AB - Completed forms of the Separation Individuation Test of Adolescence (SITA) and the Millon Adolescent Personality Inventory were obtained from 117 clinical subjects. In addition, clinicians who were working with the subjects rated them on the SITA's subscale dimensions. The results were compared with those previously found for a nonclinical population. Moderately strong internal structural properties were attained for six of the nine SITA subscales. Personality external criteria correlations showed a great deal of convergence with findings previously obtained from a nonclinical sample and were consistent with the theoretical foundations of the SITA subscales. Clinician ratings did not tend to correlate with the complementary SITA subscales, although they did with SITA subscales that were conceptually similar. PMID- 8408677 TI - Eating Disorders Inventory-2: cognitive-behavioral dimensions with nonclinical adolescents. AB - The EDI-2 was administered to 122 nonclinical females aged 12 to 18 years. Although group means were within the normal range, 30% of the subjects were abnormally preoccupied with body proportions, and 4% were pathologically weight fixated. Three risk groups were identified on the basis of high composite factor scores: (a) the Psychological Distress group indicated substantial apprehension about social adequacy, personal intimacy, emotional security, and anger control; (b) the Body-image Dysphoria group were all preoccupied with weight and body proportions at levels commensurate with diagnosed anorexics; and (c) the Behavioral Disparity group endorsed unrealistically high standards for behavior and feelings of virtue/power through behavioral restraint, yet a willingness to engage in binging and/or purging behaviors. PMID- 8408678 TI - Reliability of the Turkish version of the Perceived Social Support from Friends and Family scales, Scale for Interpersonal Behavior, and Suicide Probability Scale. AB - This study investigated the reliability of the Turkish version of the Perceived Social Support (PSS) from Friends (PSS-Fr) and Family (PSS-Fa) scales, the Scale for Interpersonal Behavior (SIB), and the Suicide Probability Scale (SPS). Both test-retest and internal consistency reliability estimates were computed. Highly adequate reliability coefficients are recorded for the subscales and the total scales of the PSS, SIB, and SPS. The results of the present study clearly support the use of Turkish versions of the PSS-Fr, PSS-Fa, SIB, and SPS as reliable methods for assessing perceived social support from friends and family, assertiveness, and suicide risk. PMID- 8408679 TI - A factor analytic study of coping styles and the MMPI-2 content scales. AB - The relationships among coping styles and psychopathology were examined with the MMPI-2 content scales and a reliable and valid coping measure--the Coping Inventory For Stressful Situations (CISS). Subjects were 167 higher-functioning normal adult males. The results are consistent with previous research that employed the CISS; a strong positive association was found between emotion oriented coping strategies and various measures of psychopathology. Task-oriented coping strategies were found to be unrelated to these measures. Results are discussed in the context of a potential model for understanding the various relationships among coping styles and psychopathology. PMID- 8408680 TI - Hispanic-white MMPI comparisons: does psychiatric diagnosis make a difference? AB - Recent research on Blacks or African-Americans in psychiatric settings suggests that Black-White MMPI differences are minimized or eliminated when researchers match or control for key moderator variables, including psychiatric diagnosis. This study attempted to determine whether Hispanic-White MMPI differences are either minimized or eliminated when Hispanics and Whites are matched on three key moderator variables--age, education, and psychiatric diagnosis. Results suggest that even after controlling for these variables, MMPI differences between Hispanics and Whites are evident. Recommendations for culturally sensitive Hispanic MMPI research are presented. PMID- 8408681 TI - Comparison of Wechsler vs. Mayo summary scores in a clinical sample. AB - Normative bases for the Wechsler Adult Intelligence Scale-Revised (WAIS-R) and the Wechsler Memory Scale-Revised (WMS-R) recently were extended through age 97 with the publication of norms that permit computation of MAYO IQs for the WAIS-R and MAYO Memory Indices for the WMS-R. This study compares traditional Wechsler summary scores with their corresponding MAYO values in a clinical sample. MAYO summary scores are correlated highly with traditional Wechsler values, and concordance rates are also strong. Despite their similarities MAYO summary scores and traditional Wechsler values are not interchangeable. Cautions are discussed relative to the use of both MAYO and traditional Wechsler IQs and Memory Indices. PMID- 8408682 TI - Therapist construct systems in use during psychotherapy interviews. AB - An innovative psychotherapy process research methodology that combines Interpersonal Process Recall with Repertory Grid analysis is introduced. This methodology was employed to examine the construct systems used by a therapist to make client behaviors meaningful. The therapist's construct systems varied across the three interviews studied. In the interview during which the therapist was judged to be most effective, the construct system used evidenced a higher level of cognitive complexity than the construct systems used in the other two interviews. Research applications of the methodology are discussed. PMID- 8408683 TI - Beyond the interpersonal circumplex in group psychotherapy: the structure and relationship to outcome of the Individual Group Member Interpersonal Process Scale. AB - Development of a new scale to measure patient behavior in group psychotherapy, the Individual Group Member Interpersonal Process Scale (IGIPS), is described. This scale captures clinically important aspects of how patients act in group, including how they respond to the other group members. The IGIPS was applied to videotapes of the first four sessions of seven 15-session outpatient therapy groups (52 patients) in the Mental Health Department of a health maintenance organization. The scale was found to have five factors (Activity, Interpersonal Sensitivity, Comfort with Self, Self-Focus, and Psychological Mindedness). Patient behavior that was moderate on two IGIPS dimensions was connected with better outcome than was that of patients with more extreme behavior in either direction. PMID- 8408684 TI - The effects of written autobiographical recollection induction procedures on mood. AB - This study assessed the effects of group induction procedures that are practical in their administration (written format) and also individualized. Fifty-four females and 36 males were assigned randomly to one of three conditions. Conditions One and Two consisted of subjects being asked to think of the two saddest or two happiest events of their lives, respectively. Condition Three consisted of a control condition in which subjects were asked to read a geography article. The procedure produced marked decreases in depression (p < .001) and anxiety (p = .001) as mood states in Condition One (happy events) and marked increases in depression (p < .001) and anxiety (p < .001) in Condition Two (sad events). These procedures are particularly suitable for mood induction in a group setting. PMID- 8408685 TI - MMPI profiles of child abusers. AB - One hundred eighty-three adults who were confirmed abusers or neglecters of children served as subjects. Their MMPI profiles were examined with respect to sex of perpetrator, type of maltreatment (i.e., sexual, physical, neglect) and offending status (perpetrator or passive perpetrator). Passive perpetrators included those individuals who did not abuse children, but had knowledge of the abuse and allowed it to occur. Results indicated no significant differences for males or females across any of the MMPI scales relative to type of abuse or offending status. However, data suggested that active perpetrators were generally more disturbed than passive perpetrators, with profile elevations consistent with a characterological configuration. PMID- 8408686 TI - Personality characteristics of self-mutilating male prisoners. AB - Self-mutilating behavior (SMB) in prisons has long been recognized as a problem. MMPI data were obtained from 30 mutilating and 30 non-mutilating male inmates. Analyses of MMPI scores revealed significant differences on nine of the clinical and validity scales. Mutilators also had more frequent elevations over 70. Interpretation of scale and subscale configurations indicates that mutilators have more somatic complaints, subjective distress, alienation, inmature defenses, and acting out tendencies than controls. This is consistent with descriptions of SMB in the literature. SMB is conceptualized as a form of aggression in a population of impulsive and alienated individuals in a high-stress environment. The frustration-aggression model is proposed as a model for understanding and further investigating this phenomenon. PMID- 8408687 TI - Issues and concerns in master's-level training and employment. AB - The controversy that surrounds Master's-level training in psychology prompted a study of the training and professional activities of those with subdoctoral training. Individuals with a Master's degree in psychology and agencies that employ them were surveyed independently to ascertain the duties performed by and assigned to them. Response rates of 21% and 24% for individuals and agencies, respectively, resulted from the mailing. The results suggested a discrepancy between the projections of agencies and individuals about the need for services in the future. Both agencies and individuals reported a substantial amount of time spent in direct services, therapy, or psychological testing. While individuals generally were satisfied with their career decision, they were less so with the decision to seek licensure as a psychological examiner. PMID- 8408688 TI - Correlates of the Woodcock-Johnson Reading Comprehension and Kaufman Brief Intelligence Test in a forensic psychiatric population. AB - Assessing the forensic psychiatric patient remains an ongoing challenge. This study assessed the intellectual status of lower-functioning incarcerated adult males with the Woodcock-Johnson Reading Comprehension subtest and the Kaufman Brief Intelligence Test. The results of this study have led the authors to argue for a broad-based assessment model that focuses on the demands and constraints of the dispositional setting. PMID- 8408689 TI - Near patient testing: the challenge for clinical pathology. PMID- 8408690 TI - Biliary tract pathology in patients with AIDS. PMID- 8408691 TI - Diagnosis of intestinal and disseminated microsporidial infections in patients with HIV by a new rapid fluorescence technique. AB - AIMS: To assess the value of a new rapid fluorescence method for the diagnosis of microsporidiosis in HIV seropositive patients. METHODS: Microsporidian spores in stools were demonstrated by using the fluorochrome stain Uvitex 2B. The new technique was evaluated in three groups of HIV seropositive patients with diarrhoea. Group 1: 19 patients with biopsy confirmed E bieneusi infection (186 stool samples); group 2: 143 consecutive patients from whom faeces were submitted for routine investigation of diarrhoea (318 samples); group 3: 16 patients with small intestinal biopsy specimens negative for microsporidia (55 samples). The new method was used to monitor spore shedding during experimental treatment with paromomycin and albendazole in four patients. RESULTS: Brightly fluorescent spores were detected in all stool samples of patients in group 1. In group 2 16 (11%) patients had spores in their stool samples. E bieneusi was found in 11 patients; in the other five another genus of microsporidia, Encephalitozoon, was recognised. Encephalitozoon spores were also found in the urine of three of these patients and in the maxillary sinus aspirate of two of them, suggesting disseminated infection. The results were confirmed by electron microscopic examination. In group 3 negative biopsy specimens were confirmed by negative stool samples in all cases. Treatment with albendazole and paromomycin did not affect the spore shedding in three patients with E bieneusi infection. By contrast, in a patient with Encephalitozoon sp infection albendazole treatment resulted in clinical improvement together with complete cessation of spore excretion in the stool. CONCLUSION: The Uvitex 2B fluorescence method combines speed, sensitivity, and specificity for the diagnosis and treatment evaluation of intestinal and disseminated microsporidiosis. PMID- 8408692 TI - Negative cytology preceding cervical cancer: causes and prevention. AB - AIM: To assess the validity of negative cervical smear reports in women who subsequently developed cervical cancer; and to determine means of improving the screening process. METHODS: One hundred and forty cervical smears, initially reported as negative from 103 women, and taken up to 12 years before diagnosis of cervical cancer, were reviewed. RESULTS: Ninety two smears contained dyskaryotic cells. Analysis showed that these smears formed several well defined patterns. False negative reports were likely to occur if fragments of neoplastic tissue rather than dissociated dyskaryotic cells were present or if the smear contained few dyskaryotic cells. Screening fatigue appeared to be a factor in others. It was also considered important that smears contained cells from the endocervix. These were deficient in 64% of the 47 smears confirmed as negative on review and in 69% of smears containing only a few dyskaryotic cells. CONCLUSIONS: Current methods of quality assurance will not remedy these defects in the screening process. It is the responsibility of laboratories to identify sources of poor smears and liaise with smear takers to ensure an improvement in quality. Assessment of the quality of smears received by a laboratory should become an important part of audit. Staff training should place more emphasis on the interpretation of "microbiopsies". The adoption of a quick scanning technique before conventional screening would probably also substantially reduce false negative results. PMID- 8408693 TI - Immunohistochemical study of tissue factor expression in normal intestine and idiopathic inflammatory bowel disease. AB - AIMS: To investigate the localisation of tissue factor expression in normal and inflamed intestine. METHODS: Serial cryostat sections of tissue taken from patients with Crohn's disease (n = 8), ulcerative colitis (n = 5), and from controls (n = 5) were stained with haematoxylin and eosin and immunostained for tissue factor, collagen type IV, fibrinogen and platelet glycoprotein IIIa. RESULTS: In control tissues tissue factor was present as a continuous layer along the epithelial basal lamina: sections from controls did not immunostain for fibrinogen or platelets. In non-ulcerated inflamed mucosa, tissue factor staining intensified in cases of Crohn's disease and was associated with fibrin deposition. Staining for tissue factor was either patchy or absent in cases of ulcerative colitis and there was no fibrin deposition. This change accompanied the early destruction of the epithelial basal lamina in ulcerative colitis that was not seen in Crohn's disease. In both diseases tissue factor expression in severely inflamed and ulcerated mucosa was present on lamina propria macrophages and vascular endothelium and was associated with fibrin or platelet thrombi. In three of eight cases of Crohn's disease tissue factor expression and thrombi were evident in areas of submucosal vasculitis. These were not seen in adjacent normal vessels. CONCLUSIONS: These observations are consistent with a tissue factor haemostatic barrier in the intestine: this barrier seems to be incomplete or defective in ulcerative colitis. Tissue factor expression by macrophages and endothelial cells may be important, particularly in the microvascular thrombosis and induration which are characteristic of Crohn's disease. PMID- 8408694 TI - Strawberry gums: a clinicopathological manifestation diagnostic of Wegener's granulomatosis? AB - AIMS: To highlight an uncommon but characteristic gingival lesion associated with Wegener's granulomatosis, emphasising the presence of pseudoepitheliomatous hyperplasia, microabscesses, and multinucleate giant cells; and the paucity of the currently accepted histopathological criteria of Wegener's granulomatosis- namely necrosis, vasculitis, and granulomata. METHODS: The histopathological features of a gingival biopsy specimen from a case of Wegener's granulomatosis limited to the upper aerodigestive tract in a 36 year old woman were compared with those of 14 similar reported cases. RESULTS: Pseudoepitheliomatous hyperplasia, microabscesses, and multinucleate giant cells were recorded as present in almost all cases of gingival Wegener's granulomatosis. Necrosis, vasculitis, and granuloma formation were present in only a few cases. CONCLUSIONS: The combination of pseudoepitheliomatous hyperplasia, microabscesses, and multinucleate giant cells in a gingival biopsy specimen from erythematous, swollen gums, clinically resembling over-ripe strawberries, in a patient with a severe systemic upset, is so typical of Wegener's granulomatosis as to be virtually diagnostic. Recognition of this characteristic lesion may aid early diagnosis and treatment in cases where other diagnostic variables are absent. PMID- 8408695 TI - Human papillomavirus DNA in glandular lesions of the uterine cervix. AB - AIMS: To assess the role of human papillomavirus in the pathogenesis of adenocarcinoma in situ, endocervical glandular dysplasia (a presumed precursor of adenocarcinoma) and endocervical glandular epithelial giant cell change. METHODS: Viral detection was carried out using an in situ hybridisation technique on paraffin wax sections. Biotinylated probes for human papillomavirus types 6/11, 16/18, 31/33/35 were used with a colorimetric detection system. RESULTS: Nine out of 21 (43%) cases of adenocarcinoma in situ contained human papillomavirus types 16/18, one of which was also positive for 31/33/35. Ten cases of glandular dysplasia and four cases of glandular epithelial multinucleation did not react with the probes used. CONCLUSIONS: These results indicate that while adenocarcinoma in situ is strongly associated with human papillomavirus infection, endocervical glandular dysplasia and glandular epithelial multinucleation are probably not associated with the virus. PMID- 8408696 TI - Is high AgNOR quantity in hepatocytes associated with increased risk of hepatocellular carcinoma in chronic liver disease? AB - AIMS: To evaluate whether high numbers of silver staining nucleolar organiser regions (AgNORs) in hepatocytes are associated with increased risk of hepatocellular carcinoma in chronic liver disease. METHODS: The quantitative distribution of AgNORs was studied in the liver biopsy specimens of 33 patients with chronic liver disease, 11 of whom developed hepatocellular carcinoma. The interval between liver biopsy and diagnosis of hepatocellular carcinoma was 26 months (range one to 61 months); the mean follow up of patients without hepatocellular carcinoma was 45 months (range 24-59 months). Quantitative evaluation of AgNORs was carried out on silver stained routine sections by morphometric analysis, using a computer assisted image analysis system. RESULTS: High interphase AgNOR values (> 3 microns2) were found in hepatocytes of nine out of the 11 (82%) patients in whom neoplastic transformation occurred. Of the remaining 22 patients, only seven (31%) had AgNOR values higher than > 3 microns2 (chi 2 4.83; p = 0.036). CONCLUSIONS: These results indicate that high numbers of interphase AgNORs are associated with increased risk of hepatocellular carcinoma in patients with chronic liver disease. PMID- 8408697 TI - Familial hypercholesterolaemia: pilot study to identify children at risk. AB - AIMS: To evaluate a more effective method of identifying children with familial hypercholesterolaemia by screening a population at high risk. METHODS: Domiciliary measurement of random cholesterol concentration was made in 200 children who were first or second degree relatives of subjects with premature onset coronary artery disease. Measurements were taken by a health visitor using a portable analyser. RESULTS: Twelve new cases of familial hypercholesterolaemia were identified during the first nine months of the study. Random cholesterol concentrations were within the normal range (< 5.2 mmol/l) in 70.5% of samples tested. Forty two (21%) of patients tested had a borderline cholesterol (5.2-5.9 mmol/l) but 50% of these fell within the normal range when fasting capillary samples were analysed. Children with significant hypercholesterolaemia on random testing (concentrations of > 5.9 mmol/l) (8.5%) also had fasting venous blood assayed for high density lipoprotein (HDL) cholesterol and tri-glyceride in the laboratory. Results indicated that 6.5% of patients screened were at high risk of cardiovascular disease (ratio of total: HDL cholesterol of > 4.5), and 1% had a moderately increased risk (ratio 3.5-4.5). CONCLUSIONS: Children with familial hypercholesterolaemia can be identified from a selected "high risk" population by measuring random capillary cholesterol concentration. PMID- 8408698 TI - Biochemical detection of phaeochromocytoma: should we still be measuring urinary HMMA? AB - AIMS: To compare the diagnostic value of biochemical tests in the detection of phaeochromocytoma. METHODS: Urinary catecholamines and metabolites were measured by high performance liquid chromatography in the initial 24 hour collections from 31 patients with histologically confirmed phaeochromocytoma. Results were compared with values from 50 patients investigated for the possible presence of a phaeochromocytoma but in whom an alternative diagnosis was later established. RESULTS: The diagnostic sensitivity for the measurement of normetadrenaline (NMT) (97%) was greater than any other single factor. Use of a combined noradrenaline and adrenaline value in preference to individual values increased the sensitivity of free catecholamines to 97%. Urinary 4-hydroxy-3-methoxymandelic acid (HMMA) showed a much lower sensitivity for the detection of phaeochromocytoma (81%). An increased excretion of either noradrenaline, adrenaline, or combined catecholamines was found in all 31 patients. CONCLUSIONS: A combination of biochemical tests improves the detection of phaeochromocytoma. The measurement of urinary free catecholamines or metadrenalines, or both, is better than HMMA estimation. It is recommended that the practice of using only HMMA measurements for the biochemical detection of phaeochromocytoma should be abandoned. PMID- 8408699 TI - Benign familial hyperphosphatasaemia as a cause of unexplained increase in plasma alkaline phosphatase activity. AB - AIMS: To consider a possible genetic origin for the persistent unexplained increase in plasma alkaline phosphatase (ALP) in five non-related patients referred over an 18 month period. METHODS: Plasma ALP isoenzyme activities were measured in patients and their first degree relatives. RESULTS: In each patient there was a noticeable increase in intestinal plasma ALP, either alone or accompanied by an increase in bone or liver ALP. Family studies showed an unexpected increase in plasma ALP and similar isoenzyme changes in first degree relatives. The findings were consistent with autosomal dominant inheritance. CONCLUSION: Inherited raised plasma ALP activity is a reasonably common cause of persistent unexplained hyperphosphatasaemia which deserves wider recognition. PMID- 8408700 TI - Cryopreservation of red blood cells: effect of freezing on red cell quality and residual lymphocyte immunogenicity. AB - AIMS: To investigate treatment with glycerol/washing as a potential substitute for freeze-thawing in the production of leucocyte depleted red cell concentrates for patients with a history of non-haemolytic reactions following transfusion. METHODS: The standard procedure of treatment with glycerol/-80 degrees C freezing/thawing/washing was compared with a similar procedure in which freezing was omitted. The quality of the resulting red cell products was assessed in relation to: (1) standard red cell biochemical parameters; (2) leucocyte and lymphocyte subset composition using flow cytometry with fluorescent labelled monoclonal antibodies; and (3) immunogenicity of the residual lymphocytes in mixed lymphocyte culture. RESULTS: Compared with red cells subjected to the standard freeze-thaw technique, red cells undergoing the non-freezing procedure and suspended in additive solutions had significantly better biochemical preservation after 21 days of storage (p < 0.001). Both procedures removed an average 98% of the initial leucocytes at the expense of 18-20% of the red cells. The non-freezing procedure resulted in higher residual concentrations of HLA class II bearing lymphocytes (p < 0.01), but not higher numbers of dendritic cells. Both procedures were equally effective in annulling the residual lymphocytes' ability to act as stimulator cells in one-way mixed lymphocyte culture. CONCLUSIONS: The non-freezing procedure produces a superior product for the provision of red cells to patients with granulocyte antibodies. These products may also offer a lower risk of HLA alloimmunisation to previously unexposed patients. PMID- 8408701 TI - Bacteriological evaluation of a down-draught necropsy table ventilation system. AB - AIMS: To evaluate the microbiological efficacy of a down-draught necropsy table ventilation system (which surrounds the cadaver with a "curtain" of air under continuous extraction) during post mortem procedures. METHODS: Air sampling was carried out both in the presence and absence of staff and cadaver and during a full post mortem procedure, with functioning and non-functioning table air extraction. The penetration of the air "curtain" was also examined during the use of an oscillating bone saw by means of a tracer organism, Bacillus subtilis var niger, painted on to the skull. RESULTS: There was little difference between bacterial counts obtained in the presence of staff only, staff plus cadaver, or during a post mortem examination. With all counts obtained, however, there was a two to three-fold reduction when the ventilation was in operation compared with when the extract duct was occluded. Using the tracer organism, a two to three log reduction in counts was shown when the "curtain" was in operation during the use of the oscillating bone saw. CONCLUSIONS: These results suggest that the system provides potential protection for post mortem room staff against airborne infections. PMID- 8408702 TI - Enhanced cultivation of Helicobacter pylori in liquid media. AB - AIMS: To evaluate a technique for culture of Helicobacter pylori in large quantities of liquid media and to determine the factors that could influence the results. METHODS: Fifteen clinical isolates of H pylori and a reference strain of H pylori NCTC11637 were used to evaluate a method to cultivate the organism in 100 ml liquid medium comprising brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Tissue culture flasks containing the inoculated liquid medium were placed in a CO2 incubator with 5% CO2 for 2 hours and then incubated in a shaking incubator at 120 rpm. RESULTS: All the clinical isolates and the reference strain grew in the broth, although only a moderate growth of the reference strain occurred. Inoculum size significantly influenced the kinetics of growth of H pylori in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B, used to suppress contamination, did not affect growth of H pylori in the medium. CO2 was essential for H pylori to grow or survive in the liquid medium. Incubation with CO2 in a CO2 incubator for 30 minutes or 2 hours did not affect the results. CONCLUSIONS: H pylori can be cultivated in large quantities of brain heart infusion broth with 5% horse serum and 0.25% yeast extract. Initial inoculum concentrations influence the kinetics of H pylori growth in the liquid medium. Vancomycin, nalidixic acid, and amphotericin B can be used as selective antimicrobial agents. CO2 is essential for initial growth of H pylori in liquid media. The findings in this study may provide a useful, reproducible, and simple method for biochemical, molecular, and physiological studies of H pylori, when those require large quantities of the organism. PMID- 8408703 TI - Distribution of Helicobacter pylori colonisation and associated gastric inflammatory changes: difference between patients with duodenal and gastric ulcers. AB - AIMS: To determine the gastric distribution of Helicobacter pylori in patients with duodenal and gastric ulcers; and to examine the mucosal inflammatory response. METHODS: Patients with newly diagnosed, uncomplicated duodenal and gastric ulcers were endoscoped and two biopsy specimens each taken from the antrum and the body. Specimens were evaluated blind by one pathologist to determine H pylori activity (scored 0-3) and inflammatory changes (according to the Sydney classification). RESULTS: Adequate biopsy material was obtained from 40 and 44 patients with gastric and duodenal ulcers, respectively. Although antral colonisation with H pylori was more common in the antrum of the latter, the organism was equally likely to be found in the body of both sets of patients; the density of colonisation was higher in those with gastric ulcers. Active gastritis and mucosal atrophy were more common in the body of those with gastric ulcers; intestinal metaplasia was also more common in the antrum of these patients. CONCLUSIONS: Gastritis in patients with duodenal ulcers is mainly antral, but the incidence of gastric body colonisation with H pylori seems to be the same in patients with either type of ulcer. There is, however, a significant difference in colonisation density. The cause and importance of this are not obvious and may be related to either host or organism factors. PMID- 8408704 TI - Cytokines in stools of children with inflammatory bowel disease or infective diarrhoea. AB - AIMS: To determine the concentrations of interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF alpha) in stools from children. METHODS: Stool samples from 14 healthy children, 32 children with inflammatory bowel disease, and 23 children with acute diarrhoea were emulsified in an equal volume of phosphate buffered saline and then centrifuged to produce a clear supernatant fluid. IL-6 and TNF alpha were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: TNF alpha was detected in the stools of all 14 healthy children (12-130 pg/g stool), but IL-6 was detected only in three. Similar results were seen in children with inactive inflammatory bowel disease. Stool TNF alpha concentrations were raised in samples from children with active inflammatory bowel disease, but in most (11/18) of these samples IL-6 was undetectable. Stool samples contained a heat labile factor which rapidly destroyed IL-6 immunoreactivity. Most children with diarrhoea had TNF alpha concentrations similar to those of healthy controls and most were also negative for IL-6. Three children with Shigella flexneri infection had extraordinarily high concentrations of both TNF alpha and IL-6 in their stools. CONCLUSIONS: There is constant low grade production of TNF alpha in the intestine of healthy people. Raised values are indicative of mucosal inflammation, but are not specific. Stool IL-6 is of little use in assessing mucosal inflammation because immunoreactivity is rapidly lost in stool samples. PMID- 8408705 TI - Interaction of coagulase negative staphylococci with lectins. AB - AIMS: To investigate the reaction of 118 blood culture isolates of coagulase negative staphylococci with a panel of seven lectins. METHODS: The interactions between the bacterial suspensions and lectins from Arachnis hypogaea (peanut agglutinin), Bauhina purpurea, Solanum tuberosum (potato starch), Triticum vulgaris (wheat germ agglutinin), Wisteria floribunda, Concanavalin ensiformis and Limulus polphemus (horse-shoe crab agglutinin) were assayed in microtitre plates incubated for 1 hour at room temperature then left overnight at 4 degrees C. Agglutinating activity was detected by examining the pattern of cell settlement compared with that of the controls. RESULTS: Lectins from Solanum tuberosum, Triticum vulgaris, Concanavalin ensiformis and Limulus polyphemus agglutinated 90% of the strains and displayed 11 agglutination patterns which were unrelated to species, clinical relevance, or antibiotic resistance. Fifty three per cent of the isolates fell into three reaction patterns and the other patterns were represented by nine or fewer strains. Replicate cultures investigated simultaneously gave consistent results, but some strains exhibited variation in agglutination patterns on repeat testing. CONCLUSIONS: Based on these observations lectin agglutination patterns seem to offer a method with potential for strain differentiation among coagulase negative staphylococci. Reproducibility may be improved by the use of biotinylated lectins and growing the coagulase negative staphylococci on defined media. Discrimination can be increased by the inclusion of other reactive lectins. PMID- 8408706 TI - Acute myelomonocytic leukaemia following atypical congenital rubella. AB - The child of a woman immunised against rubella presented at 5 months with developmental delay and recurrent infection; she was shown to have congenital rubella. At 15 months she developed acute myelomonocytic leukaemia (AMML). Rubella is difficult to diagnose after immunisation. AMML has not been previously described in association with congenital rubella, as far as is known. PMID- 8408707 TI - Combined silver Perls's stain for differential staining of ringed sideroblasts and marrow iron. AB - During a study of nucleolar organiser regions, a modified silver stain was found to be a sensitive marker for the iron in ringed sideroblasts, more so than Perls's stain when the marrow iron stores were low. To enhance the usefulness of the silver stain, a combined silver Perls method was developed. This stains the ringed sideroblast iron black and haemosiderin blue, thus rendering the detection of ringed sideroblasts easier even when marrow iron stores are excessive. AT the same time, it allows marrow iron content to be evaluated. The silver reagent in this combined method probably shows phosphate rather than the iron present in the abnormal mitochondria in ringed sideroblasts. This facilitates the differential staining of ringed sideroblast "iron" and haemosiderin. PMID- 8408708 TI - Pseudotumoral amyloidosis of beta 2-microglobulin origin in the buttock of a patient receiving long term haemodialysis. AB - A 52 year old man who had been receiving haemodialysis for 13 years, with a history of renal tuberculosis, right ischial tuberculous osteomyelitis, and dialysis arthropathy, developed a soft tissue tumour in his left buttock. Histological analysis, immunohistological staining, and electron microscopic examination of the surgically removed tumour showed massive deposits of beta 2 microglobulin (beta 2-M) amyloid. This case shows the expanding clinical spectrum of this type of amyloidosis, and it is suggested that amyloid infiltration should be considered in the differential diagnosis of gluteal tumours in these patients. PMID- 8408709 TI - Diagnosis of childhood BK virus cystitis by electron microscopy and PCR. AB - A case of BK virus cystitis in a 5 year old boy is reported. This patient, who was not immunocompromised, had had acute cystitis for two weeks. Many intracytoplasmic inclusions were observed in urinary sediment smears stained by the Papanicolaou method. Electron microscopic examination showed virus particles, presumed to be human polyomavirus, in the nuclei of the degenerated urothelial cells. A DNA sequence of the BK virus was detected in 200-300 urothelial cells in Papanicolaou stained smears by the polymerase chain reaction. BK virus is an unusual cause of symptomatic cystitis in a healthy child. PMID- 8408710 TI - Frozen section simulation of trabecular adenoma and medullary cancer by papillary thyroid carcinoma. AB - Two patients are reported in whom initial misdiagnoses of medullary cancer or trabecular hyaline adenoma for papillary carcinoma were made. PMID- 8408711 TI - Rearrangements of c-myc and c-abl genes in tumour cells in Burkitt's lymphoma. AB - Rearrangements of oncogenes c-myc and c-abl were detected by non-radioactive hybridisation in a case of Burkitt's lymphoma/leukaemia. The surface phenotype of Burkitt's cells were positive for CD19, CD20, HLA-DR, CD14, CD33 and surface immunoglobulin markers. Although cytogenetic analysis was not performed, the c myc and heavy immunoglobulin genes had the same 14.2 kilobase EcoRI molecular size fragment, suggesting a possible t(8;14) translocation which is a common marker of this malignancy. The c-abl oncogene was also rearranged in DNA digested BamHI and EcoRI. The physiopathological implications of the rearranged c-abl gene are unknown, this being the first case, as for as is known, of Burkitt's lymphoma/leukaemia with a rearranged c-abl gene. PMID- 8408712 TI - Prospective comparative study of computer programs used for management of warfarin. PMID- 8408713 TI - Vasculopathy and antiphospholipid antibodies in systemic lupus. PMID- 8408714 TI - Relative friendly death certificates. PMID- 8408715 TI - Relative friendly death certificates. PMID- 8408716 TI - Application of the community periodontal index of treatment needs (CPITN) in a group of 45-54-year-old German factory workers. AB - The purpose of the present study was to assess the periodontal status of 45-54 year-old patients and to evaluate their treatment needs. Probing depths, bleeding on probing and retentional elements (calculus and overhanging restorations) were determined according to the community periodontal index of treatment needs (CPITN). Additionally, loss of attachment was measured. Results indicated that none of the subjects had a completely healthy periodontium; only 14.7% presented with single sextants which were healthy or needed only improved oral hygiene. Slightly less than half (46.1%) of the subjects were classified as treatment need (TN) category 2 and the remainder (53.9%) as TN3. Of the subjects classed as TN category 3, 14% had the requisite code 4 in one sextant, 18.2% in 2 sextants, 21.7% in half or more of the sextants and 4.2% in all sextants. With a mean of 5.55 sextants per patient, 0.2 sextants per person were scored as code 0 or 1, 1.33 sextants as code 2, 2.79 sextants as code 3 and 1.24 sextants as code 4. The mean loss of attachment was 3.8 mm. Anterior teeth showed less loss of attachment than posterior teeth and buccal and lingual surfaces showed less loss of attachment than mesial and distal surfaces. The data indicate that although this group of 45-54-year-old subjects had high CPITN scores in total TN categories, the codes for complex Treatment Needs (TN3) were recorded only in localized areas. PMID- 8408717 TI - Antimicrobial properties of human dentin impregnated with tetracycline HCl or chlorhexidine. An in vitro study. AB - Substantivity of tetracycline HCl and chlorhexidine digluconate to human root dentin was assessed in vitro. 51 extracted single-rooted teeth, their crowns removed, were assigned to 1 of 4 treatments in groups of 12. A control groups included 3 roots. Each group was divided into 3 subgroups to allow evaluation of drug exposure for 1, 3 or 5 min. The roots were immersed in tetracycline HCl (10 or 50 mg/ml) or chlorhexidine digluconate (0.12 or 0.2%) solutions following root planning. Control roots were immersed in sterile saline (0.9%). Following drug immersion, the roots were transferred to tubes containing 2 ml tris buffered saline. The tubes were incubated at room temperature for 22 days. Desorption media were replaced at 24-h intervals. Removed media were examined for antimicrobial activity using a microtiter assay in which bacterial growth was evaluated by optical density readings. Roots immersed in tetracycline HCl 50 mg/ml released antimicrobial activity to successive desorption media for 14 days. Tetracycline HCl 10 mg/ml activity lasted 4 days. Roots subjected to chlorhexidine digluconate released antimicrobial activity for 24 h only. Within each treatment, there were no differences between the 3 exposure intervals of 1, 3 or 5 min. Our findings suggest usage of the periodontally exposed instrumented root as a depot for sustained release of tetracycline HCl, but not chlorhexidine digluconate, to the subgingival environment. The substantiveness of tetracycline HCl seems related to drug concentration rather than the exposure interval. Clinical trials are needed to confirm the clinical significance of these in vitro observations. PMID- 8408718 TI - Elevated opsonic activity for Porphyromonas (Bacteroides) gingivalis in serum from patients with a history of destructive periodontal disease. A case: control study. AB - We have measured the opsonic capacity of serum for the phagocytosis of Porphyromonas (Bacteroides) gingivalis by polymorphonuclear leucocytes (PMN) in 35 patients with a history of destructive periodontitis and 35 matched control subjects. The serum from cases, tested at concentrations of 8% and 0.8% opsonised P. gingivalis for phagocytosis by PMN to a level significantly greater than controls (p < 0.0001 and < 0.01 respectively). IgG antibody levels to P. gingivalis whole cells estimated by ELISA were also significantly higher in the cases (p < 0.0001). The IgG antibody levels correlated significantly with the opsonic capacity of the serum tested at 8% concentration in controls (r = 0.371, p = 0.03) but not in cases (r = 0.235, p = 0.17); in 0.8% serum, the opsonic capacity of the cases and controls were not significantly correlated. Elevated opsonisation by serum was a significant predictor that a subject was a case rather than a control, even after allowing for the effect of elevated IgG antibody in the cases. The data suggest that an elevated capacity of serum to opsonise P. gingivalis is a distinctive feature in patients with past destructive periodontal disease. PMID- 8408719 TI - Microcirculatory dynamics in experimental human gingivitis. AB - The purpose of this study was to determine the changes that occur in the gingival microcirculation during the development of experimental gingivitis in humans. There have been no studies published to date combining videomicroscopy and laser Doppler flowmetry to study vascular dynamics in experimental gingivitis. Alterations occurring in the microcirculation of the marginal gingiva in 10 (18 30-year-old), healthy male humans when they suspended oral hygiene procedures in a proscribed area for 12-16 days were monitored. A partial mouth, experimental gingivitis model was employed. Gingival health was evaluated before and after the experimental period by assessing gingival and plaque indices and gingival crevicular fluid volume. Gingival vascular monitoring included measurement of red blood cell velocity in individual gingival microvessels via videomicroscopy and measurement of regional gingival blood flow using laser doppler flowmetry. The number of vessels visible in a given microscopic field in a given subject and the number of vessels exhibiting flow were also determined from the videotapes. Systemic cardiovascular and respiratory parameters were monitored to ensure that gingival vascular changes were not secondary to systemic changes. Gingivitis developed in all subjects; significant increases (Student t-test, p < 0.05) were seen in plaque index, gingival index, bleeding on probing and crevicular fluid volume. No change in superficial capillary blood velocity and a significant decrease in gingival regional blood flow were seen with gingivitis. A significant increase in the number of vessels visible in microscopic fields and a decrease in the % of vessels exhibiting flow were observed. Gingival microcirculation exhibited a dramatic, dynamic change in response to the development and progression of gingivitis. PMID- 8408720 TI - A socio-ecologic model for periodontal diseases. AB - A socio-ecologic conceptual model for periodontal diseases has been proposed. The model, which includes 4 items: health-care organization, human biology, behavioral factors, and environment, has been tested on a random-sample of 50 year-old Oslo citizens. The investigation is based on a clinical examination as well as a questionnaire and structured interview. A logistic regression model was used to study associations between risk factors and the probability of deep periodontal pockets (> 5.5 mm). The risk of periodontal pockets was positively associated with: short duration of education, being of male gender, previous periodontal treatment, poor oral hygiene and infrequent toothbrushing. Variables describing behavioral factors and human biology were the items found to be most closely associated with periodontal pocketing. PMID- 8408721 TI - Mechanical stimulation by toothbrushing increases oxygen sufficiency in human gingivae. AB - We studied the immediate effect of mechanical stimulation with a toothbrush on oxygen sufficiency in human gingivae. Teeth were brushed with a modified Bass technique at various forces (100, 200, and 300 g) for various times (5, 10, and 30 s). Hemoglobin (Hb) oxygen saturation was measured before and intermittently for 60 min after brushing, by non-invasive tissue reflectance spectrophotometry. Hb oxygen saturation increased within a few min after brushing and then slowly returned to the initial level. Toothbrushing at 200 g for 10 s caused the longest lived increase in Hb oxygen saturation; saturation remained about 6% above the baseline level for more than 25 min. Before brushing, Hb oxygen saturation was significantly lower in inflamed gingivae than in healthy gingivae (p < 0.05). After brushing of inflamed gingivae, the Hb oxygen saturation almost reached the level measured in healthy gingivae before brushing. These findings suggest that stimulation with a toothbrush increases oxygen sufficiency in both healthy and inflamed gingivae. PMID- 8408722 TI - Digital radiographic image-based bone level measurements: effect of film density. AB - Previous studies have demonstrated that accurate alveolar bone-level measurements can be made using digitally enhanced bitewing radiographs. In this study, we determined the effect of exposure variations on bone-level measurement accuracy using digital radiographic images of dry skulls. 6 direct bone-level measurements on the mandibular 1st molars of dry skulls were established by consensus of 2 expert observers. 13 bitewing radiographs were taken on each side of 11 skulls, using a specially designed positioning device. Exposure settings of 70 kVp, 10 mA, and 24 impulses were empirically chosen as the baseline. Kilovoltage and impulses times were individually varied (50 to 100 kVp in 10 kV increments and 15 to 48 impulses in 6 steps) to ensure a wide range of clinically relevant optical densities on the test images. The radiographs were digitized and alveolar bone level measurements, corresponding to those for skulls, were made on the images by 2 observers. Pearson product-moment coefficients between radiographs and criteria measurements were calculated. It was found that dry skull and digital bitewing radiographic measurements were highly correlated, within the limits of exposure parameter variations tested. Accurate measurements of alveolar bone level are possible from bitewing radiographs taken within a wide range of exposures, when digital image processing techniques are employed. PMID- 8408723 TI - Measures of treatment efficacy. AB - Measures of treatment efficacy are those numbers we think about when we decide whether one treatment is "better" than another. Such measures quantify the differences between treatments and help patients and clinicians make informed choices. The usual measure of treatment efficacy in periodontal research has been the mean difference between treatments in probing level measures. This measure has frustrated clinicians and researchers alike for its failure to communicate the size of the association between treatment and clinical outcome. How does one interpret the clinical relevance of a small mean difference between treatments, such as 0.4 mm? This report compares the advantages and disadvantages of the different measures of treatment efficacy: the mean difference, the relative risk, significance levels (P-values), the risk difference, and, the preventable fraction. PMID- 8408724 TI - Excretion of lignocaine and its metabolite monoethylglycinexylidide in breast milk following its use in a dental procedure. A case report. AB - The excretion of lignocaine in breast milk has been documented in a 34-year-old woman following the injection of 20 mg lignocaine for a dental alloy restoration in the right upper quadrant. Lignocaine and its primary metabolite monoethylglycinexylidide in milk and plasma were quantified by high-performance liquid chromatography. The concentration of lignocaine in milk ranged from 44-66 micrograms l-1 while that for monoethylglycinexylidide ranged from 35-41 micrograms l-1. The milk: plasma ratios for lignocaine and monoethylglycinexylidide were 1.1 and 1.8, respectively. The calculated daily infant doses for the parent drug and metabolite were both less than 0.01 mg kg-1 day-1. With the exception of very rare allergic reactions, these levels of infant exposure are extremely low and of no toxicological significance. Nursing mothers receiving lignocaine for standard dental procedures can be advised that continuation of breast feeding is safe. PMID- 8408726 TI - Big science little science: science in controversy. PMID- 8408725 TI - A dose-response study of triclosan mouthrinses on plaque regrowth. AB - Triclosan is used in toothpastes and mouthrinses as a plaque inhibitory agent. The concentrations used and therefore the dose of triclosan varies between products and there is, as with most plaque inhibitory agents, little information on the dose response of this agent. The purpose of this investigation was to measure the plaque inhibitory properties of triclosan in a simple mouthrinse formulation over a range of concentrations and therefore doses. The study was a 5 treatment, double-blind, Latin-square randomised, minus-active controlled design balanced for residual effects. The formulations were 0.01%, 0.05%, 0.1% and 0.2% triclosan in 0.5% sodium carbonate and 5% alcohol aqueous solutions, used as 10 ml, 1-min rinses 2 x daily, without tooth-brushing. 2 groups each of 15 healthy volunteers rinsed during alternate 4-day treatment, 1-week washout periods with the allocated rinses. From a zero-plaque baseline on Day 1, plaque was scored by index and area on Day 5. A dose response pattern of decreasing plaque indices and particularly areas, with increasing triclosan dose was observed. However, by far the largest sequential drop in plaque scores occurred between 0.1% and 0.2% triclosan treatments. Extending the dose-response study to higher concentrations is considered impractical if not unjustifiable, because of potential harmful local side-effects and compliance difficulties. Dose-response studies to assess the agents thought to potentiate triclosan would seem warranted. PMID- 8408727 TI - The phenomenon of abusable psychotropic use among North American youth. AB - Abusable psychotropic use can, and does, affect all North American youth, either directly or indirectly, regardless of age, gender, culture, ethnic background, education, race, or socioeconomic status. Over the last decade, the morbidity and mortality associated with abusable psychotropic use among youth have become staggering. A current overview of the phenomenon of abusable psychotropic use among youth in North America, including the use of alcohol, cannabis, cocaine, LSD, nicotine, and polyabusable psychotropic use, is presented with attention to the expanding role of clinical pharmacologists in relation to both prevention and treatment. The Mega Interactive Model of Abusable Psychotropic Use Among Youth is presented as a heuristic device to assist clinical pharmacologists, and other health care providers, in addressing the multifactorial interactive aspects of this complex phenomenon as observed in the pediatric age group. In this regard, attention is given to the interaction of the Abusable Psychotropic Dimension, including the Abusable Psychotropic Variables (e.g., pharmacokinetics, abuse potential) and Pattern of Use Variables (e.g., social use, abuse, compulsive use), with the Young Person, Societal, and Time Dimensions. PMID- 8408728 TI - The disposition of doxorubicin on repeated dosing. AB - Twelve cancer patients (aged 49-74 years) receiving doxorubicin (66 +/- 8 mg/m2) as a 1-hour intravenous infusion had serial serum samples (0-48 hours) obtained after the first and second courses of therapy. Mean number of days between courses was 24.3, and all patients had normal liver function. Patients received the same concomitant antineoplastic agents and doses in both courses. Doxorubicin and doxorubicinol concentrations were assayed by high-performance liquid chromatography and fitted to a two- or three-compartment infusion model. White blood cell and platelet toxicity were evaluated as (initial--nadir/initial)* 100. Differences in pharmacokinetic parameters were determined by a paired t test. Wide intrapatient and interpatient variability was seen between therapeutic courses. A significant decrease in the apparent volume of distribution of the central compartment (Vc = 16.6 versus 10.4 L/m2; P < .05), and a nonsignificant decrease in clearance (CL = 748 versus 658 mL/min/m2) was observed on the second course of therapy. Doxorubicinol area under the curve and elimination half-life were similar between courses. Extensive chemotherapy-induced changes in white blood cell and platelet counts were observed but were similar in degree for courses 1 and 2. These data suggest that higher initial doxorubicin concentrations on the second course of therapy are secondary to an alteration in distribution volume (Vc). In this subset of patients, however, these changes were not associated with an increase in hematologic toxicity. PMID- 8408729 TI - Disposition of olsalazine and metabolites in breast milk. AB - This study examined the disposition of olsalazine and its metabolites into breast milk after the ingestion of a single dose of 500 mg olsalazine. Blood and serum samples were obtained for 48 hours after the ingestion of 500 mg olsalazine in a 39-year-old lactating woman. Blood samples were obtained at .0, .5, 1, 2, 4,6, 24.5, 26, and 48 hours. Maternal milk samples were obtained at .0, .5, 2, 4, 6, 14, 24, 28, 36, and 48 hours. Olsalazine and olsalazine-S underwent high-pressure liquid chromatography analysis, and 5-ASA and Ac 5-ASA underwent fluorometric detection. Acetylated-5-ASA achieved concentrations of .8, .86, and 1.24 mumol/L in breast milk at 10, 14, and 24 hours, respectively. Olsalazine, olsalazine-S, and 5-ASA were undetectable in the breast milk for 48 hours after drug administration. Clinically significant drug exposure in the breast-fed infant is unlikely after a maternal single dose of olsalazine. Idiosyncratic hypersensitivity, however, remains a possibility even if the infant is exposed to only minute quantities. PMID- 8408730 TI - Use of reflectance spectrophotometry in the human corticosteroid skin blanching assay. AB - A reflectance spectrophotometric method for evaluation of the skin blanching response to topical corticosteroids was evaluated. This blanching response is used, for drug development and regulatory purposes, to assess potency and bioequivalence of topical corticosteroid products. The common method involves the use of a human rater to measure blanching response in the skin. This study evaluated an instrumental alternative to the human rater and used this method to measure the differences between a number of brand name and generic topical corticosteroid products (six creams and six ointments). Products were applied to the forearms of normal volunteers and the blanching responses were assessed after 6 and 16 hours in both occluded and non-occluded skin sites. Only the fluocinolone acetonide generic and brand name preparations were different from each other. The spectrophotometric method proved to be equivalent but not superior to the standard human observer method. PMID- 8408731 TI - Localization of drug release sites from an oral sustained-release formulation of 5-ASA (Pentasa) in the gastrointestinal tract using gamma scintigraphy. AB - Release of 5-ASA from a sustained release formulation (Pentasa, Ferring A/S, Copenhagen, Denmark) was monitored with plasma sampling for up to 24 hours in nine volunteers under both fasted and fed conditions. Drug absorption was correlated with location of the sustained-release microgranules in the gastrointestinal tract by gamma scintigraphy. Disintegration of the labeled tablet preparation occurred in the stomach within 20 minutes and acetylated 5-ASA was detectable in the plasma less than 60 minutes after ingestion. No significant differences were detected in either transit times through the small intestine, peak plasma acetylated 5-ASA concentration or lag time to absorption between fasted and fed individuals. Peak plasma concentration of acetylated 5-ASA usually occurred when the microgranules were present in the small intestine or ascending colon. The pharmacoscintigraphic study confirmed that 5-ASA release from the formulation occurred throughout the gastrointestinal tract, and that food effects on the in vivo behavior of the preparation were minimal. PMID- 8408732 TI - Single- and multiple-dose pharmacokinetics of clarithromycin, a new macrolide antimicrobial. AB - The pharmacokinetics of clarithromycin and its active 14(R)-hydroxy metabolite were evaluated after single and multiple oral doses of 250 and 500 mg of clarithromycin. Multiple-dose regimens used 12-hour dosing intervals for 7 doses. Plasma and urine concentrations were measured using high-performance liquid chromatography. Appearance of clarithromycin and its metabolite in plasma were rapid, as reflected by mean times to maximum plasma concentration ranging from 1.8 to 2.6 and 1.8 to 2.9 hours, respectively. The rises in clarithromycin peak plasma concentration (Cmax) and area under the plasma concentration versus time curve (AUC) were disproportionate to increase in dose, suggesting nonlinearity in parent compound pharmacokinetics. Clarithromycin terminal disposition half-life (t1/2) also exhibited dose dependency, ranging from harmonic means of 2.7 to 4.8 hours. In contrast, based on Cmax AUC, and predicted/observed accumulation ratios, nonlinearity in metabolite pharmacokinetics was not observed. Plasma accumulation of metabolite occurred to a much lesser degree than that of the parent compound despite a substantially longer t1/2 for the metabolite (metabolite accumulation ratios based on AUC dose 7/AUC dose 1:250-mg regimen = 1.03 +/- 0.33, 500-mg regimen = 0.81 +/- 0.29, parent accumulation ratios: 250-mg regimen = 1.64 +/- 0.47, 500-mg regimen = 1.65 +/- 0.69). This would suggest that formation of this metabolite is capacity-limited and that this may in part account for the nonlinearity observed in clarithromycin pharmacokinetics. Urinary excretion constituted a relatively important route of elimination of clarithromycin, with renal clearance accounting for 17 to 31% of apparent total body clearance. PMID- 8408733 TI - Evaluation of the pharmacokinetic and pharmacodynamic interaction between quinidine and nifedipine. AB - Quinidine and nifedipine appear to be subject to metabolism by the same isozyme of cytochrome P-450. In addition, both drugs have been reported to alter the pharmacokinetics of other compounds. To investigate a potential interaction, 10 healthy subjects (five male, five female) received quinidine sulfate (200 mg orally), nifedipine (20 mg orally), or the combination of both drugs every 8 hours for 4 doses using a randomized, cross-over study design with a 2-week washout period between treatments. Drug concentration, heart rate, and mean arterial pressure were measured at frequent intervals after the final dose. Quinidine concentrations were unchanged by the co-administration of nifedipine. Nifedipine area under the curve (AUC0-8) increased 36.6% from 333 to 455 micrograms.hr/L (P < .05) after quinidine administration. Heart rate was significantly higher in the nifedipine-quinidine treatment at 0.5, 1.0, 1.5, and 2.0 hours when compared with either drug alone. The maximum increase in heart rate (17.9 beats/minute) occurred at 0.5 hours after nifedipine administration and was significantly correlated with serum concentrations at that time (r = .78). These results suggest that quinidine inhibits nifedipine metabolism, and this pharmacokinetic interaction results in enhanced pharmacologic response. PMID- 8408734 TI - Renal effects of a nonhypotensive i.v. dose of felodipine. AB - To evaluate the natriuretic effect of a nonhypotensive dose of felodipine, 11 healthy volunteers (age: 21-28 years) on a high-sodium diet received the drug or its vehicle in a double-blind, randomized, crossover study. Administered intravenously at a dose level of 7.5 micrograms/min for 30 minutes followed by 5 micrograms/min for 120 minutes, felodipine increased natriuresis (546 +/- 69 vs. 454 +/- 39 mumol/min, P < 0.001) and diuresis (8.9 +/- 0.6 vs. 7.5 +/- 0.5 mL/min), compared to its vehicle. Renal plasma flow tended to be augmented, but there was a significant reduction of renal vascular resistance (0.085 +/- 0.004 vs. 0.101 +/- 0.012 mm Hg/mL/min, P < 0.03). The glomerular filtration rate was slightly decreased and proximal sodium reabsorption was diminished with no measurable effect on distal function. Felodipine stimulated plasma renin activity, but produced no changes in plasma atrial natriuretic factor, cGMP, aldosterone, and atrial vasopressin levels. In conclusion, felodipine induced natriuresis and diuresis while reducing proximal tubular sodium reabsorption. PMID- 8408735 TI - The effect of nifedipine on the pharmacokinetics and dynamics of diltiazem: the preliminary study in normal volunteers. AB - To evaluate the influence of nifedipine on the pharmacokinetics and the pharmacodynamics of diltiazem, five healthy subjects received 60 mg diltiazem orally on two occasions, diltiazem alone or after nifedipine pretreatment (10 mg three times daily for 3 days). After nifedipine pretreatment, the maximum concentration (Cmax) of diltiazem was increased and the time of Cmax was shortened, and the area under the concentration curve (AUC) tended to be increased. Although heart rate was increased, the corrected PQ interval tended to be prolonged after the nifedipine pretreatment. Both a decreased hepatic clearance and an increased bioavailability of diltiazem probably accounts for the increase in the Cmax and AUC of diltiazem after nifedipine pretreatment, and that might affect the pharmacodynamics of diltiazem. PMID- 8408736 TI - Horizontal component of electro-oculogram as a parameter of arousal in dementia: relationship between intellectual improvement and increasing arousal under pharmacotherapy. AB - Although nootropics and metabolically active compounds sometimes attenuate intellectual deficits of dementia patients, this mechanism has not been discussed or investigated. The authors hypothesized that these compounds act to enhance attention and/or concentration by increasing the arousal level, and then improve intellectual functions. Based on this hypothesis, the authors investigated the arousal level and the improvement of intellectual functions in ten patients with Alzheimer type dementia and ten patients with multi-infarct dementia before and after 3 months pharmacotherapy using bifemelane hydrochloride. They then analyzed whether the increases in arousal level involves or relates to the improvement of intellectual functions. On the 20 patients, psychological tests were carried out, and the arousal level was evaluated using the frequency of rapid eye movements in the electro-oculogram (EOG) before and after 3 months administration of bifemelane hydrochloride. As a result, a significant positive correlation was observed between the improvement on the Hasegawa's dementia scale and the rate of increase in rapid eye movements in the EOG. The EOG monitoring can be a great help in establishing an objective evaluation method on drug efficacy. PMID- 8408737 TI - Dose-related antagonism of the emetic effect of morphine by methylnaltrexone in dogs. AB - Opioids administered to produce analgesia cause unwanted emesis in patients (incidence 20%-30%, depending on situation). Tests in animals show that quaternary narcotic antagonists like methylnaltrexone (MNTX) do not affect the analgesic potency of morphine, but such compounds have not been examined for their potential to antagonize morphine-induced emesis. To determine the effects of MNTX on emetic response, we assigned 85 dogs to one of 11 groups challenged with morphine alone or morphine and various doses of MNTX IM or i.v. Antagonism of the emetic response was dose related: MNTX, 0.25 mg/kg IM or 0.2 mg/kg i.v., completely blocked the emetic effect of morphine in dogs for approximately 60 minutes. If morphine-induced emesis is mediated by receptors available to a quaternary antagonist (perhaps on the peripheral side of the blood-brain barrier), MNTX may prevent opioid-induced emesis. These data indicate that opioid induced emesis might be prevented without affecting analgesia. PMID- 8408738 TI - Differences of chronopharmacokinetic profiles between propranolol and atenolol in hypertensive subjects. AB - Previous studies have shown that the absorption rate of a lipophilic, but not hydrophilic, agent is faster after the night dosage than after the morning dosage in nocturnal rodents. The present study examines whether such a difference in chronopharmacokinetic profiles between lipophilic and hydrophilic agents also exists in humans. Propranolol (20 mg), a lipophilic beta-blocker, or atenolol (50 mg), a hydrophilic beta-blocker, was given orally to 13 hypertensive patients at 9:00 AM (day trial) or 9:00 PM (night trial) by a crossover design. Plasma concentrations of propranolol and its metabolites, 4-hydroxypropranolol and naphthoxylactic acid, and atenolol were determined just before and at 0.5, 1, 1.5, 2, 3, 4, 6, 12, and 24 hours after treatment. Maximum plasma concentration (Cmax) and area under the plasma concentration-time curve (AUC) of propranolol in the day trial were significantly greater than those in the night trial. Time to maximum plasma concentration (tmax) was significantly shorter in the day trial. No significant difference was observed in the elimination half-life between the two trials. There were similar administration time-dependent changes in the Cmax for 4-hydroxypropranolol and naphthoxylactic acid. On the other hand, although the Cmax of atenolol was greater and its tmax was shorter in the day trial, the differences did not reach significance. These results suggest that propranolol, but not atenolol is absorbed more rapidly after the morning dosage than after the night dosage. Based on these findings, the authors speculate that the absorption rate of a lipophilic, but not hydrophilic, agent is faster after the morning dosage than after the night dosage in humans. PMID- 8408739 TI - An integrated design for dose comparison trials. AB - An integrated design, which incorporates the dose titration scheme into the parallel comparative design, is proposed for dose comparison trials of drugs that may cause a first-dose phenomenon. This design includes a concurrent placebo control group, thereby providing valid estimate of drug effect. The other groups are defined by the maximum allowable dose. Except for the maximum allowable dose, both the titration schedule and the titration interval are standardized. The effect of the pace of titration is thus controlled. In extreme cases, all patients need only the lowest dose tested, and all patients need the highest dose tested to achieve the required efficacy response. In nonextreme cases, this design answers questions that are usually asked of dose comparative trials: overall drug effects, adequacy of starting dose, effects of dose increment, maximum effective dose, dose-response relationship, and time effect. Because both efficacy and safety analyses can be performed similarly, risk-benefit analysis thus can be evaluated in the same group of patients, and an optimum titration regimen may be determined rationally. PMID- 8408740 TI - Twelve months around the world ... as a volunteer dentist. PMID- 8408741 TI - Minimum instrumentation for conservative operative procedures. PMID- 8408743 TI - The other faces of dentistry. PMID- 8408742 TI - Xylitol: a sugar that fights tooth decay. PMID- 8408744 TI - African universities as partners in community health development. PMID- 8408745 TI - Three models to promote syphilis screening and treatment in a high risk population. AB - Three syphilis control models targeted at a high incidence area were implemented and evaluated over a 31-month period. These models consisted of (1) street-based outreach and referral to community health clinics for free sexually transmitted disease screening and treatment, (2) off-site syphilis screening at high risk locations and referral for free treatment, and (3) off-site syphilis screening and referral at high risk locations with monetary incentives offered for obtaining test results and seeking treatment if required. Off-site screening- conducted at homeless shelters, soup kitchens, drug treatment centers, and community fairs--was found to be significantly more effective in promoting syphilis screening than outreach and referral to community health centers. For the off-site screening models, monetary incentives were related to an increase in the proportion of individuals at homeless shelters and soup kitchens who agreed to be tested, and to an increase in the proportion of people at homeless shelters, soup kitchens and drug treatment centers who sought their test results and obtained treatment, if required. PMID- 8408746 TI - Alcohol consumption patterns and related problems: results of a county survey. AB - This study describes alcohol use in the general population of San Diego County. A random digit dial telephone survey was utilized to query 1656 adults about their drinking behavior. A quantity and frequency measure of alcohol consumption was used to classify drinking patterns. Alcohol problems were classified by acute problems and chronic problems. Males 18-25 years old reported the highest percentages of heavier drinking. Among females, this age group was also the most likely group to report heavier drinking. Overall, males and heavier drinkers were the most likely groups to report problems. However, lighter and moderate drinkers accounted for equal or greater percentages of each problem. Drinkers in the 18-25 year old age group were the group most likely to report the majority of all problems in both problem categories. Additionally, this group accounted for the majority of the reported acute problems. PMID- 8408747 TI - A study of satisfaction among primary health care patients in Saudi Arabia. AB - Primary Health Care is essential health care based on delivering integrated health services (curative and preventive). The Kingdom of Saudi Arabia adopted this approach in 1980, and by the year 1987 the Ministry of Health had established 1477 Primary Health Care centers. The expansion in Primary Health Care created a need for various types of evaluation. Theorists recommended the study of patients' satisfaction as a way of evaluating care. The aim of this study was to assess the satisfaction of patients with different aspects of Primary Health Care services in Riyadh. The sample consisted of 300 patients chosen systematically from three Primary Health Care centers in Riyadh. The data were collected by personal interviews. The tool consisted of demographic data, a 4-point rating scale of 40 statements measuring satisfaction with different aspects of Primary Health Care services, and an open question eliciting the patients' suggestions for improvements. The analysis of variance (ANOVA) was used to determine the difference in level of patient satisfaction between the three centers. The results show that the patients were moderately satisfied with the services. They were most satisfied with the effectiveness and humane aspects of care, and least satisfied with the thoroughness and continuity aspects of care. It is recommended that the Ministry of Health develop programs for its personnel to sensitize them to the different aspects of Primary Health Care. PMID- 8408748 TI - Effects of a city ordinance regulating smoking in restaurants and retail stores. AB - The purpose of this study was to assess the effects of a city smoking ordinance on local businesses. Representatives from 61 randomly selected restaurants and retail stores in Flagstaff, Arizona were interviewed to determine their perceptions of the ordinance's effect on their business. Eighty-seven percent of the respondents were business owners or managers. Gross sales data 12 months prior to and 12 months after enactment of the ordinance were also collected for two categories of retail stores. The vast majority of restaurant and store respondents believed that the ordinance had no effect on their business. Gross sales increased an average of 16 to 25.8 percent per store during the year following prohibition of smoking in retail stores. This study did not support the belief that city smoking ordinances have a negative effect on local businesses. PMID- 8408749 TI - HMO development in an academic medical center: the rise and fall of a prepaid health program in New York city. AB - Through a documented case study the authors identify the critical factors that impede the introduction of prepaid medical care as part of education and practice within a prestigious and well established academic medical center. The inherent conflicts between individual fee-for-service practice and population-based prepaid practice and the resistance to innovations in medical care organization as they surfaced in that center, are presented. The need for a clear understanding of the complexities of HMO development and of an appreciation for the importance of a planning process in which all interested parties are involved, is emphasized. A clear commitment by policy makers, administrators and providers is highlighted as fundamental for the implementation of a system where practitioners are motivated to assume responsibility for the comprehensive care of a defined population that prepays for their services. The rewards as well as the difficulties for institutionalizing commitment to this form of health care delivery and impacting on medical education are discussed. PMID- 8408750 TI - The prevalence of ocular disorders among Hispanic and Caucasian children screened by the UCLA Mobile Eye Clinic. AB - A cross-sectional study was performed to describe the prevalence of ocular abnormalities among six and seven year old children of Hispanic and Caucasian ancestry in Los Angeles County. Data were obtained from vision screenings of lower to middle income Hispanic and Caucasian children completed by the UCLA Mobile Eye Clinic at public neighborhood elementary schools and community centers between January and August of 1989, and January and March of 1990. Out of a total sample of 854 children, 64% were Hispanic; 36% were Caucasian; 51% were female and 49% were male. Ocular abnormalities observed were similar for both ethnic groups. Caucasians, however, showed non-significant, but consistently higher prevalences of most ocular abnormalities. Stronger associations between ethnicity and visual abnormalities were observed within the female subgroup. For example, hyperopia was found more commonly among female Caucasian children than among female Hispanic children (p < 0.01). This information can be used in planning for the eye care needs of communities with Caucasian and Hispanic components. PMID- 8408751 TI - A baseline assessment of cigarette sales to minors in San Diego, California. AB - This study reports the sales rate of cigarettes to minors in San Diego County, and investigates factors associated with cigarette sales to minors. Two hundred and ninety-four stores were identified and recruited to participate in a retailer education effort. To assess the baseline illegal sales rate of cigarettes to minors, selected stores were surveyed by 70 volunteer teams of adults and minors. Questionnaires were also administered to participating store owners or managers to assess retailers' knowledge about laws regulating minors' access to tobacco. Survey results indicated that minors were able to successfully purchase cigarettes in 68% of attempts. In addition, teen gender, community sociodemographics and cashier characteristics were associated with sales to minors. Analyses of the retailer questionnaire indicated retailers knew the legal age to purchase cigarettes, but few knew of the specific penalties associated with sales to minors. These findings indicate that minors have easy access to cigarettes and underscore the need for intensive tobacco sales education for retailers and enforcement of sales to minors laws. PMID- 8408752 TI - Alcohol use and potential risk for alcohol-related adverse drug reactions among community-based elderly. AB - This paper documents the frequency of alcohol consumption and concurrent use of alcohol and medications in a random sample of elderly community dwellers. Further, a profile of older persons who are likely to be drinking alcohol is developed and the extent to which they are at potential clinical risk due to their concurrent use of alcohol with prescription and over-the-counter medications is explored. While approximately 43 percent are abstainers, the majority of older respondents reported using alcohol. Older drinkers who take one or more drugs which place them at potential risk for negative drug-alcohol interactions represent one-quarter of this sample but are often overlooked in estimating the extent of alcohol problems in the elderly. By far, the most common risk was from the use of OTC pain medications and alcohol (19 percent). The multivariate analyses revealed that sex, educational attainment, and religious affiliation are important factors to consider in developing a profile of older people who are at risk for alcohol-related ADRs. Implications for health care and social service professionals who work with elderly community-dwellers are discussed. PMID- 8408753 TI - Rural hospital health promotion: programs, methods, resource limitations. AB - Most hospitals provide health promotion programs for community residents. There is little information concerning the specific types of services offered by rural hospitals. A questionnaire was sent to every acute care hospital in Iowa (N = 124), including 99 rural hospitals and 25 urban hospitals. Surveys were returned from 95 rural hospitals (96%) and 20 urban hospitals (80%). Results indicated that 98.9% of rural hospitals offered health promotion services to community residents. These services provided on average 7.5 programs on a regular basis, while using only 1.2 full-time equivalent (FTE) employees. Urban hospitals provided 9.5 regular programs with 2.4 FTE. The most common types of rural promotion programs were blood pressure screening, cholesterol screening, safety and protection programs, diet/nutrition programs, prenatal/maternal health, and breast cancer screening. Over 40% of rural respondents stated that other less common programs, including substance abuse prevention and mental health promotion, were needed but could not be offered because of resource limitations; these types of services were offered more commonly in urban hospitals. Rural hospital health promotion programs are attempting to meet a wide variety of programming needs with limited resources, and attention may be well directed towards finding how best to provide various programs with limited resources to maximize their impact on community health. PMID- 8408754 TI - Comparison of the immunocytochemical localization of DARPP-32 and I-1 in the amygdala and hippocampus of the rhesus monkey. AB - Dopamine and adenosine 3':5'-monophosphate (cAMP) regulated phosphoprotein of M(r) 32 kDa (DARPP-32) and phosphatase inhibitor 1 (I-1) have been associated with intracellular signal transduction processes and share several biochemical features. Localization of each phosphoprotein in distinct neural structures will aid investigation of their physiologic properties and help identify their unique roles in the nervous system. We have compared the distribution of the two phosphoproteins in the amygdala and hippocampus of the rhesus monkey with the aid of immunocytochemical procedures. Neurons immunoreactive to antibodies raised against the phosphoproteins DARPP-32 and I-1 were noted in the cortical, central, and components of the basal group, including the basomedial, the lateral, and to a lesser extent, the basolateral amygdaloid nuclei. Within the large basal nuclei positive neurons were found preferentially in their medial and ventral subdivisions. By making a direct comparison in the same animals, we observed differences in the distribution of the two phosphoproteins in the amygdala. DARPP 32 and I-1 positive neurons overlapped partially in the basal nuclei, to a lesser extent in the cortical, but were segregated in the central amygdaloid nucleus with neurons positive for DARPP-32 noted laterally, and for I-1 medially. In contrast to the amygdala, where numerous DARPP-32 and I-1 positive neurons were observed, only I-1 had a notable presence in the hippocampus. Moreover, I-1 associated label was found only in neurons in the granule cell layer of the dentate gyrus, their dendritic plexus, and axons which innervate hilar and CA3 neurons. DARPP-32 and I-1 are intracellular messengers associated with signal transduction. Their regional distribution in the amygdala and the hippocampus suggests an involvement in the level of excitability of specific components of these limbic structures. Moreover, our results suggest that I-1 has a unique role in the intrinsic circuitry of the hippocampal formation and indicate a system where the physiologic properties of I-1 may be studied in isolation. PMID- 8408755 TI - Cortical connections of inferior temporal area TEO in macaque monkeys. AB - In macaque monkeys, lesions involving the posterior portion of the inferior temporal cortex, cytoarchitectonic area TEO, produce a severe impairment in visual pattern discrimination. Recently, this area has been shown to contain a complete, though coarse, representation of the contralateral visual field (Boussaoud, Desimone, and Ungerleider: J. Comp. Neurol. 306:554-575, '91). Because the inputs and outputs of area TEO have not yet been fully described, we injected a variety of retrograde and anterograde tracers into 11 physiologically identified sites within TEO of seven rhesus monkeys and analyzed the areal and laminar distribution of its cortical connections. Our results show that TEO receives feedforward, topographically organized inputs from prestriate areas V2, V3, and V4. Additional sparser feedforward inputs arise from areas V3A, V4t, and MT. Each of these inputs is reciprocated by a feedback projection from TEO. TEO was also found to have reciprocal intermediate-type connections with the fundus of the superior temporal area (area FST), cortex in the most posteromedial portion of the superior temporal sulcus (the posterior parietal sulcal zone [area PP]), cortex in the intraparietal sulcus (including the lateral intraparietal area [area LIP]), the frontal eye field, and area TF on the parahippocampal gyrus. The connections with V3A, V4t, and PP were found only after injections in the peripheral field representations of TEO. Finally, TEO was found to project in a feedforward pattern to area TE and to areas anterior to FST on the lateral bank and floor of the superior temporal sulcus (areas TEm, TEa, and IPa, Seltzer and Pandya: Brain Res. 149:1-24, '78), all of which send feedback projections to TEO. Feedback projections also arise from parahippocampal area TH, and areas TG, 36, and possibly 35. These are complemented by only sparse feedforward projections to TG from central field representations in TEO and to TH from peripheral field representations. The results thus indicate that TEO forms an important link in the occipitotemporal pathway for object recognition, sending visual information forward from V1 and prestriate relays in V2-V4 to anterior inferior temporal area TE. PMID- 8408756 TI - Ultrastructural analysis of sympathetic neuromuscular junctions on mesenteric veins of the guinea pig. AB - This study reports on the detailed ultrastructure of sympathetic postganglionic varicose axon terminals on mesenteric veins leading from the ileum of the guinea pig and in particular the structural arrangement of the varicosities with venous smooth muscle cells. The response to nerve stimulation in veins has a long time course and it has been suggested that this reflects a wide separation between the site of transmitter release and the receptors on the effector cell membrane. The aim of this study was to determine the distance between individual sympathetic varicosities and smooth muscle cells in mesenteric veins. Fluorescent histochemical preparations of the sympathetic innervation of the different branches of mesenteric veins indicate the branching network of varicose axons around the vessel to be relatively dense. Electron micrographs show the innervation to be confined to the adventitia close to the medio-adventitial border and to be predominantly catecholaminergic. A serial section ultrastructural analysis of the relationship of the varicosities with the outer smooth muscle cells showed that almost all (98%) of the exposed axon varicosities in the adventitia formed neuromuscular junctions. Three-dimensional reconstructions from serial sections of individual varicosities have shown that the junctions have structural specialisations identical to neuromuscular junctions described on arterial vessels and similar to those found at skeletal neuromuscular junctions. The density of neuromuscular junctions on the veins was found to be similar to that on the corresponding artery in the same animal. We suggest that in veins, noradrenaline is released focally at neuromuscular junctions. PMID- 8408757 TI - Cortical hierarchy reflected in the organization of intrinsic connections in macaque monkey visual cortex. AB - Neuronal response properties vary markedly at increasing levels of the cortical hierarchy. At present it is unclear how these variations are reflected in the organization of the intrinsic cortical circuitry. Here we analyze patterns of intrinsic horizontal connections at different hierarchical levels in the visual cortex of the macaque monkey. The connections were studied in tangential sections of flattened cortices, which were injected with the anterograde tracer biocytin. We directly compared the organization of connections in four cortical areas representing four different levels in the cortical hierarchy. The areas were visual areas 1, 2, 4 and Brodman's area 7a (V1, V2, V4 and 7a, respectively). In all areas studied, injections labeled numerous horizontally coursing axons that formed dense halos around the injection sites. Further away, the fibers tended to form separate clusters. Many fibers could be traced along the way from the injection sites to the target clusters. At progressively higher order areas, there was a striking increase in the spread of intrinsic connections: from a measured distance of 2.1 mm in area V1 to 9.0 mm in area 7a. Average interpatch distance also increased from 0.61 mm in area V1 to 1.56 mm in area 7a. In contrast, patch size changed far less at higher order areas, from an average width of 230 micron(s) in area V1 to 310 micron(s) in area 7a. Analysis of synaptic bouton distribution along axons revealed that average interbouton distance remained constant at 6.4 micron(s) (median) in and out of the clusters and in the different cortical areas. Larger injections resulted in a marked increase in the number of labeled patches but only a minor increase in the spread of connections or in patch size. Thus, in line with the more global computational roles proposed for the higher order visual areas, the spread of intrinsic connections is increased with the hierarchy level. On the other hand, the clustered organization of the connections is preserved at higher order areas. These clusters may reflect the existence of cortical modules having blob-like dimensions throughout macaque monkey visual cortex. PMID- 8408758 TI - Distribution of acetylcholine receptors in the central nervous system of adult locusts. AB - A polyclonal antibody raised against nicotinic acetylcholine receptor protein from purified locust neuronal membrane was used to analyse the distribution of antigenic sites within the central nervous system of adult Schistocerca gregaria. Light microscopic examination showed that all principal neuropiles in the thoracic ganglia label with the antibody but that the major tracts and commissures do not. Analysis of this pattern of staining in the electron microscope reveals that the receptor is present on specific synaptic and extrajunctional neuronal membranes in the neuropile. Antigenic sites are also evident on the plasma membranes and within the cytoplasm adjacent to Golgi complexes of some neuronal somata, suggesting that these neurones synthesise nicotinic acetylcholine receptors. In addition to neuronal labelling, there is evidence that the receptor is also present on the membranes of three types of glial cells. The implications of this pattern of receptor distribution are discussed. PMID- 8408759 TI - Transient appearance of calbindin-D28k-positive neurons in the superior olivary complex of developing rats. AB - Calbindin-D28k (CaBP) is a calcium-binding protein that is prominent in various parts of the mammalian auditory system. In order to shed some light on the possible role of CaBP during ontogeny, when calcium ions play key roles in several processes, the location of CaBP was examined immunocytochemically in the auditory system of the developing rat. This study focuses on the principal nuclei of the superior olivary complex, which show distinct CaBP labeling in the adult. Consistent with previous reports in the rat and other mammals, CaBP immunoreactivity in adults was intense in somata of the medial nucleus of the trapezoid body (MNTB) and in the neuropil (presumably in axons) of the lateral superior olive (LSO), the superior paraolivary nucleus (SPN), and the medial superior olive (MSO). In fetal and neonatal animals, however, the labeling pattern was strikingly different. Around birth, MNTB neurons are immunonegative for CaBP, whereas somata and processes in the LSO, probably neuronal, are heavily labeled at that age. This labeling pattern persists throughout the first week of postnatal life and begins to change at P8, when MNTB neurons become immunopositive for CaBP. During the next 10 days labeling intensity in MNTB neurons increases considerably, and the increase is paralleled by an increase in labeling intensity of the neuropil in the LSO, SPN, and MSO, indicating that the labeled processes in these nuclei may be axons originating from MNTB neurons. Immunoreactivity in LSO cells begins to decline around P8, decays rapidly between P10 and P18, and reaches its adult level around P28, when the CaBP labeling pattern in the whole superior olivary complex is indistinguishable from that in the adult. The present results show that the development of CaBP immunoreactivity in the rat superior olivary complex is characterized by two reciprocally related processes: as immunoreactivity within MNTB somata and fibers in the SPN, and LSO, and the MSO increases between P8 and about P21, the immunoreactivity in LSO neurons declines. CaBP immunoreactivity in LSO neurons is only transiently present, suggesting a critical period in development during which the control of Ca2+ homeostasis in LSO neurons may be of particular importance. PMID- 8408760 TI - Developmental regulation of Fos and Fos-related antigens in cerebral cortex, striatum, hippocampus, and cerebellum of the rat. AB - Previous work has associated the proto-oncogene c-fos with such events as neuronal excitation and cell growth and differentiation. This study specifically examined the expression of the Fos protein as well as other Fos-related antigens (Fras) during postnatal development of rat brain. Ages P1 through P15 as well as adult animals (P60) were examined. Particular focus was placed on developing cerebral cortex, striatum, hippocampus, and cerebellum. We used both the Alu antiserum, which recognizes the Fos protein specifically, and the M5 antiserum, which recognizes both Fos and a family of Fos-related antigens. Fos and Fras were developmentally regulated in a region- and cell-specific manner. Differential nuclear and cytoplasmic labeling appeared age dependent. Transient Fos expression was generally followed by a more protracted time course of Fra expression. Fos and a delayed or an extended expression of Fras were observed in subplate neurons between P1 and P15, in striatal striosome and matrix neurons between P1 and P9, and in hippocampal pyramidal neurons between P1 and P9. Fras alone were expressed in cerebral cortex pyramidal neurons and other cortical neurons between ages P1 and P15. Fos and Fras were concomitantly expressed in piriform and entorhinal cortical neurons between P1 and P9 and in cerebellar Purkinje cells between ages P5 and P10. Constitutive levels of Fos and Fras remained detectable in adult animals in a subset of cerebral cortical neurons and cerebellar Purkinje neurons. PMID- 8408761 TI - Cortical modules in the posteromedial barrel subfield (Sml) of the mouse. AB - An antibody to MAP2 was used on sections through the posteromedial barrel subfield (PMBSF) of primary somatosensory cortex to reveal the distributions of cell bodies and dendrites. It was found that apical dendrites of layer VI neurons form irregular bundles or sheets that break up in layer IV, where most of these dendrites form their terminal tufts. In contrast, the apical dendrites of layer V pyramidal neurons form clusters that ascend into layer II/III where they are joined by apical dendrites of the superficial pyramidal neurons. In layer IV the clusters of the layer V apical dendrites are more concentrated in barrel walls than in hollows. Thus, in layer IV the average center to center spacing between the clusters is about 25 microns in the barrel hollows, and about 22 microns in the barrel walls. In part, this differential distribution of the apical dendritic clusters is brought about because the apical dendrites of layer V pyramids beneath the periphery of the barrel hollows arc towards the barrel walls as they pass from layer V into layer IV. Based on previous analyses of the three dimensional organization of the primary visual areas in the monkey, cat, and rat, it has been proposed that neurons in these cortices are organized into modules that are centered on the clusters of apical dendrites belonging to layer V pyramidal neurons. Mouse PMBSF cortex is composed of similar pyramidal cell modules and the organization of neurons in these modules is similar to that in visual cortex. This suggests that the pyramidal cell modules are fundamental neuronal units that exist throughout the cerebral cortex, and implies that the various functional areas of the cortex in different species are organized according to a common, basic plan. PMID- 8408762 TI - Quantitative study of primary sensory neurone populations of three species of elasmobranch fish. AB - In order to assess the ability of sharks and rays to sense pain, the proportion of myelinated versus unmyelinated sensory fibres in the dorsal roots and the diameter spectrum of cells in the dorsal root ganglia of three species of elasmobranch fish were ascertained. Electron micrographs were used to count the numbers of myelinated and unmyelinated fibres in montages of whole dorsal roots of the long-tailed stingray (Himantura sp.), the shovelnose ray (Rhinobatus battilum), and small specimens of the black-tip shark (Carcharhinus melanopterus). The diameters of dorsal root ganglion cells in each species were measured by using the light microscope. Less than 1% of the dorsal root axons in the long-tailed stringray and a large specimen of the shovelnose were unmyelinated, whereas in smaller shovelnose rays and in the small black-tipped sharks, from 14% to 38% of axons were unmyelinated. Unmyelinated fibres differed from those in mammalian nerves in that there was a one-to-one association of the fibre with a Schwann cell. We conclude from these observations that myelination was incomplete in the black-tipped sharks and the smaller specimens of the shovelnose rays. The distribution of the diameter of cells of the dorsal root ganglia of these species was unimodal, resembling the diameter range that has been reported for the somata of myelinated fibres in the cat. We interpret these results as indications that sharks and rays lack the neural apparatus essential for the sensation of pain and we suggest that, to these life forms, the perception of pain might have little relevance to survival. PMID- 8408763 TI - The dorsomedial visual area of owl monkeys: connections, myeloarchitecture, and homologies in other primates. AB - Cortical connections of the dorsomedial visual area (DM) of owl monkeys were revealed with injections of the bidirectional tracer, wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP), or the retrograde fluorescent tracer, diamidino yellow. Microelectrode recordings in two cases identified DM as a systematic representation of the visual hemifield in a densely myelinated rectangle of cortex just rostral to the dorsomedial portion of the second visual area (V-II, or area 18). Cortex was flattened and cut parallel to the surface in all cases so that the myeloarchitectonic borders of DM and other areas such as the primary visual area (V-I or area 17), V-II or area 18, and the middle temporal visual area (MT) could be readily determined, and the surface view patterns of connections could be directly appreciated. The ipsilateral pattern of connections of DM were dense and visuotopically congruent with area 17, area 18, and MT, and moderate to dense connections were with the medial visual area (M), the rostral division of the dorsolateral visual area, the dorsointermediate area, the ventral posterior area, the caudal division of inferotemporal cortex (ITc), the ventral posterior parietal area, and visuomotor cortex of the frontal lobe. The connections of DM were concentrated in the cytochrome oxidase (CO)-dense blobs of area 17, the CO-dense bands of area 18, and the CO-dense regions of MT. Callosal connections of DM were with matched locations in DM in the opposite hemisphere, and with VPP. The ipsilateral connections of DM with area 17 were confirmed by injecting WGA-HRP into area 17 in one owl monkey. In addition to labelled cells and terminals in area 18 and MT, bidirectionally transported tracer was also apparent in DM. Evidence for the existence of DM in other primates was obtained by injecting area 17 and examining the areal patterns of connections and myeloarchitecture in three species of Old World monkeys, two additional species of New World monkeys, and prosimian galagos. In all of these primates, one of three major targets of area 17 was a densely myelinated zone of cortex just rostral to dorsomedial area 18, in the location of DM in owl monkeys. Thus, it seems likely that DM is a visual area common to all primates. PMID- 8408764 TI - Central retinal area is not the site where ganglion cells are generated first. AB - The development of retinal ganglion cells (RGC) was studied in the chick from stage 18 to adulthood. Our main objectives were to identify the retinal site where the first RGCs differentiate, to locate this site relative to the optically defined central retinal area, and to map the spatial arrangement of the RGC field at different stages in development. The eyes of the experimental animals were fixed and serially sectioned. The borders of RGC fields were determined from the presence of either ganglion cell perikarya or ganglion cell axons. In seven cases between stages 21 and 26, the borders of the RGC fields were confirmed electron microscopically. The serial sections together with the RGC fields were then reconstructed in three dimensions. The reconstructed retinae were projected onto a plane by using the radially equidistant polar azimuthal projection. First, RGCs appear dorsal to the apex of the optic fissure. Ganglion cell development then initially spreads out symmetrically with respect to the optic fissure. However, from stage 29 on, the nasal half of the retina expands much more than the temporal half. This asymmetrical growth entails that the optic fissure is eventually located in the temporal half of the retina in the mature animal. The RGC fields of the embryonic stages were superimposed on the retina of a visually active animal according to their real size and position. It turned out that the central retinal area was at least 2 mm away from the site where the first RGCs were generated. It is not before stage 28 that the prospective central retinal area is included into the expanding ganglion cell field. The fact that RGCs at the central retinal area are generated 2.5 days later than first RGCs near the apex of the optic fissure has important implications for the formation of the retinotectal projection. PMID- 8408765 TI - A quantitative study of the lateral spread of Muller cell responses to retinal lesions in the rabbit. AB - A wide variety of retinal pathology is associated with an increase in Muller glial cell expression of glial fibrillary acidic protein (GFAP). In this study the time course and spatial spread of the Muller cell GFAP response following argon laser photocoagulation lesions was examined in wholemounted rabbit retina. At 24 hours single focal lesions were surrounded by GFAP positive Muller cell end feet which declined in density with distance but extended as far as 2-3 mm from the lesion. The Muller cell reaction reached a maximal spread of 4-5 mm at 14 to 21 days and had started to contract by 30 days, leaving a core of GFAP positive processes immediately around the lesion site at 60 days. This zone of spread was much larger than the area of disrupted pigment epithelium. Isodensity plots did not reveal any correlation with the trajectory of retinal ganglion cell axons. The spread of reaction was more confined for lesions within the visual streak than in the dorsal or ventral retinal periphery. Multiple lesions within a focal region of retina resulted in a greater density of GFAP reactive end feet with a corresponding greater spread. However, when five to ten lesions were made in a horizontal row, the Muller cells over the entire retina became GFAP immunoreactive. This pan-retinal reaction took several days to spread, peaked at 7-14 days, and contracted back to the primary lesion sites by 2 months. This spread of Muller cell reactivity may be triggered by the diffusion of substances released by injury or it may be due to direct cellular communication. The extensive indirect effect on Muller cells of laser irradiation might be an important component of the clinical effect of laser photocoagulation and indicates a long distance communication mechanism between retinal glia which is poorly understood. This study also shows the importance of the time at which the Muller cell response is assessed. PMID- 8408766 TI - Estrogen receptor-immunoreactive forebrain neurons project to the ventrolateral hypothalamus in female guinea pigs. AB - In rodents, the facilitation of sexual receptivity by estradiol and progesterone is suspected to be mediated by a network of neurons containing estrogen and progestin receptors. In female guinea pigs, this network would include estrogen receptor-immunoreactive (ER-ir) neurons located within the rostro-ventral ventrolateral hypothalamus (r-vVLH). This hypothesis predicts that a proportion of the neurons projecting to the r-vVLH contains estrogen receptors. This prediction was tested through retrograde tracing combined with immunocytochemistry for estrogen receptors. Retrogradely labelled neurons were particularly abundant within the medial preoptic nucleus (MPN), bed nucleus of the stria terminalis (BST), anterior hypothalamus, amygdala, and lateral parabrachial nucleus. As predicted by the hypothesis, retrogradely labelled neurons were mostly observed in estrogen receptor-rich areas. Retrogradely labelled neurons also containing estrogen receptor-immunoreactivity (ER-IR) were primarily found within the MPN, BST, and amygdala. However, a majority of retrogradely labelled neurons did not contain ER-IR. As the preoptic area and the r-vVLH are both responsive to estradiol in the facilitation of sexual receptivity by progesterone, these data are consistent with the hypothesis tested. However, our data also suggest that the network of neurons controlling sexual receptivity may include elements not directly sensitive to estradiol. Finally, the location of retrogradely labelled neurons is discussed with respect to the stimuli provided to the r-vVLH in the context of sexual receptivity facilitated by estradiol and progesterone. PMID- 8408767 TI - Distribution of prosaposin-like immunoreactivity in rat brain. AB - Prosaposin is the precursor for saposins A, B, C, and D, which are small lysosomal proteins required for the hydrolysis of sphingolipids by specific lysosomal hydrolases. With a monospecific anti-saposin C antibody, which cross reacts with prosaposin but not with saposin A, B, or D, the present immunoblot experiments showed that the rat brain expresses an unprocessed approximately 72 kDa protein (possibly prosaposin) and little saposin C. Regional analysis demonstrated that prosaposin is abundant in the brainstem, hypothalamus, cerebellum, striatum, and hippocampus, and less abundant in the cerebral cortex. Consistent with this finding, prosaposin-like immunoreactive neurons and fibers as revealed by immunohistochemistry were observed frequently in subcortical regions. The medial septum, diagonal bands, basal nucleus of Meynert, ventral striatum, medial habenular nucleus, and motor nuclei of cranial nerve had significant numbers of immunoreactive neurons. There were also nerve fibers with prosaposin-like immunoreactivity in several projection fields of the above nuclei. Other brain areas that contained prosaposin-like immunoreactive neurons and/or processes were: several brain nuclei (nucleus caudate putamen, globus pallidus, substantia nigra, red nucleus) constituting the so-called extrapyramidal system, reticular thalamic nucleus, entopeduncular nucleus, mammillary nuclei, auditory relay nuclei, cerebellum, sensory cranial nerve nuclei, and the reticular formation. The distribution pattern of prosaposin is apparently different from that of other neuroactive substances so far examined, and thus prosaposin may be involved in novel central events. PMID- 8408768 TI - Ultrastructure of serotonin-immunoreactive terminals in the core and shell of the rat nucleus accumbens: cellular substrates for interactions with catecholamine afferents. AB - The nucleus accumbens is composed of a core region involved in motor functions and a shell region implicated in emotional and motivational processes. Both of these regions receive serotonin- and dopamine-containing afferents. We examined whether the serotonin innervation or relation to catecholamine (mainly dopamine) axons in the nucleus accumbens shows common features or specializations corresponding to the noted functional differences in core and shell subregions. To address this question, we examined the ultrastructure of serotonin-containing axons and their relation to catecholamine-containing afferents in either the core or shell of the nucleus accumbens. Single coronal sections through the rat forebrain were processed for immunoperoxidase labeling of serotonin and immunogold silver labeling of tyrosine hydroxylase, the catecholamine synthesizing enzyme. Varicose processes showing peroxidase product for serotonin by light microscopy were confirmed to be axons and terminals by electron microscopy. In a quantitative analysis of serotonin-immunoreactive terminals forming one or more contacts in single sections, some common features were observed. For the core (n = 120) and the shell (n = 82), 41% formed synaptic junctions with unlabeled dendrites, 75% were in apposition with unlabeled terminals, which often formed asymmetric junctions, and 20% were in apposition with axons or terminals containing tyrosine hydroxylase. Thus, in both the core and shell of the nucleus accumbens, serotonin terminals synapse on postsynaptic neurons and are likely to modulate or be modulated by presynaptic interactions with excitatory axons forming asymmetric junctions and by catecholaminergic afferents. Marked differences in the morphology of serotonin axons were also seen in the core versus shell of the nucleus accumbens. By light microscopy, serotonin immunoreactive axons were thicker and more varicose than those found in the core. Ultrastructural analysis confirmed that, in contrast to the core, serotonin immunoreactive axons and terminals in the shell were larger in cross-sectional diameter size (0.7 micron vs. 0.3 micron). Additionally, serotonin axon terminals in the shell contained more numerous immunoreactive large dense core vesicles and more frequently formed symmetric as opposed to asymmetric contacts with dendrites. The larger size and more numerous dense core vesicles in serotonin immunoreactive terminals in the shell support the concept that serotonin or co existing neurotransmitter may be more tonically released in the shell versus core of the nucleus accumbens. PMID- 8408769 TI - Ultrastructure of synapses from the pretectum in the A-laminae of the cat's lateral geniculate nucleus. AB - We have recently shown in cats that many neurons projecting to the lateral geniculate nucleus from the pretectum use gamma-amino butyric acid (GABA) as their neurotransmitter. We sought to determine the morphology of synaptic terminals and synapses formed by these pretectal axons and the extent to which they resemble other GABAergic terminals found in the geniculate neuropil (i.e., from geniculate interneurons and cells of the nearby perigeniculate nucleus). To do this, we labeled a population of pretectal axons with the anterograde tracer Phaseolus vulgaris leucoagglutinin and analyzed the morphology and synaptology of labeled pretectal terminals in the A-laminae of the cat's lateral geniculate nucleus. The pretectal projection, which arises primarily from the nucleus of the optic tract (NOT), provides synaptic innervation to elements in the geniculate neuropil. The labeled NOT terminals are densely packed with vesicles, contain dark mitochondria, and form symmetrical synaptic contacts. These are characteristics of the F1 type of terminal, and we know from other studies that GABAergic axon terminals from interneurons and perigeniculate cells also give rise to F1 terminals. We compared our population of NOT terminals with labeled perigeniculate and unlabeled F1 terminals selected from the geniculate neuropil and found that all three populations share many morphological characteristics. Both qualitative and quantitative assessments of the pretectal terminals suggest that these are a type of F1 terminal. Most pretectal terminals selectively form synapses onto geniculate profiles that contain irregularly distributed vesicles and dark mitochondria and that are postsynaptic to other types of terminals. These postsynaptic targets thus exhibit features of another class of inhibitory, GABAergic terminal known as F2 terminals, which are the specialized appendages of geniculate interneurons. Pretectal inputs, being GABAergic, may thus serve to inhibit local interneuronal outputs. Pretectal axons also innervate the perigeniculate nucleus, in which the only targets are the other main type of inhibitory, GABAergic neurons. These results suggest that the pretectum may facilitate retinal transmission through the lateral geniculate nucleus by providing inhibition to the local inhibitory cells: the interneurons and probably perigeniculate cells. This would serve to release geniculate relay cells from inhibition. PMID- 8408770 TI - Effects of aging on the size, density, and number of rhesus monkey lateral geniculate neurons. AB - Visual abilities decline during aging, and many of these declines are due to neural changes in the retina or brain. We have begun studies of the monkey visual system to investigate the location and nature of these changes as well as to answer general questions about the effects of aging on neural structure and function. We began with the dorsal lateral geniculate nucleus (LGN) because it is the main structure through which visual information passes on the way to cortex and because the parallel parvicellular and magnocellular pathways are most easily identified and studied in the LGN. In the present experiment, we determined the sizes, densities, and numbers of LGN neurons in young-adult (5 to 12.5 years) and old (23 to 27.5 years) rhesus monkeys. The measures were corrected for tissue shrinkage, and stereological procedures were used that yield unbiased estimates. In young-adult monkeys, neurons densities were lower in the magnocellular layers (about 14,000/mm3) than in the parvicellular layers (23,000/mm3). Neuron density increased about 28% from anterior to posterior in both types of layers. There was an average of approximately 1,267,000 neurons in the parvicellular layers and 148,000 neurons in the magnocellular layers; however, there was substantial variability (1.9-fold) among five brains studied. Aging produced a statistically significant decrease in neuron density in both the magnocellular (29% average decrease) and parvicellular (41% average decrease) layers. However, there was no significant loss of neurons. Rather, the density decrease was due to a small (nonsignificant) decrease in the number of neurons combined with a small (nonsignificant) increase in LGN volume. The increase in LGN volume was due to a significant increase in neuron soma-size and proportional increase in the volume of glial cells, blood vessels, and neuropil. These results, together with those of other studies, suggest that the effects of aging on the primate visual pathway from retina through striate cortex are relatively subtle. It is possible that the major neural changes occur more centrally. Alternatively, individual differences in the effect of aging may require much larger samples or prior screening to observe consistent changes. PMID- 8408771 TI - Distribution and colocalization of choline acetyltransferase immunoreactivity and NADPH diaphorase reactivity in neurons within the medial septum and diagonal band of Broca in the rat basal forebrain. AB - NADPH diaphorase histochemistry and choline acetyltransferase immunocytochemistry were used to assess quantitatively the presence of nitric oxide synthase in the cholinergic neurons of the magnocellular basal forebrain complex. Virtually all (97%) NADPH diaphorase reactive magnocellular neurons in the medial septum and the vertical and horizontal limbs of the diagonal band of Broca were choline acetyltransferase immunoreactive, whereas only a proportion of the choline acetyltransferase immunoreactive neurons were NADPH diaphorase reactive. Thus NADPH diaphorase histochemistry identified a subpopulation of the magnocellular cholinergic neurons. Occasionally, NADPH diaphorase reactive neurons were observed within the medial septum and diagonal band of Broca that were not choline acetyltransferase immunoreactive, and in general were morphologically distinct from the magnocellular neurons; such neurons are probably representatives within the medial septum and diagonal band of more widely distributed phenotypically distinct populations of NADPH diaphorase reactive neurons. The proportions of the neurons in which choline acetyltransferase and NADPH diaphorase colocalized in the medial septum and in the diagonal bands of Broca were similar in any one coronal section, but there was a considerable difference in the proportions throughout the rostrocaudal extent of these nuclei. In the most rostral sections of the medial septum and diagonal band, approximately 70% of the choline acetyltransferase immunoreactive neurons were NADPH diaphorase reactive, whereas the proportion decreased progressively to about 30% at the level of the decussation of the anterior commissure. To examine further the extent of colocalization throughout the magnocellular basal forebrain complex, sections of the magnocellular preoptic nucleus, substantia innominata, and nucleus basalis magnocellularis were examined. While there was little total colocalization of choline acetyltransferase immunoreactivity and NADPH diaphorase reactivity in any particular section (approximately 18%), almost all of the double labelled neurons were in the substantia innominata, with very few in the other nuclei. Thus although there is a caudal to rostral gradient of the proportion of magnocellular cholinergic neurons that are NADPH diaphorase reactive throughout the entire basal forebrain magnocellular complex, subregions, such as the substantia innominata and magnocellular preoptic nucleus, may not follow this trend. The recent demonstration that the NADPH diaphorase histochemical reaction localizes a nitric oxide synthase suggests that attention should be given to the NADPH diaphorase subpopulation in pathological and experimentally induced alterations of the basal forebrain. PMID- 8408772 TI - The early development of thalamocortical and corticothalamic projections. AB - The early development of thalamocortical and corticothalamic projections in hamsters was studied to compare the specificity and maturation of these pathways, and to identify potential sources of information for specification of cortical areas. The cells that constitute these projections are both generated prenatally in hamsters and they make reciprocal connections. Fluorescent dyes (DiI and DiA) were injected into the visual cortex or lateral geniculate nucleus in fixed brains of fetal and postnatal pups. Several issues in axonal development were examined, including timing of axon outgrowth and target invasion, projection specificity, the spatial relationship between the two pathways, and the connections of subplate cells. Thalamic projections arrive in the visual cortex 2 days before birth and begin to invade the developing cortical plate by the next day. Few processes invade inappropriate cortical regions. By postnatal day 7 their laminar position is similar to mature animals. By contrast, visual cortical axons from subplate and layer 6 cells reach posterior thalamus at 1 day after birth in small numbers. By 3 days after birth many layer 5 cell projections reach the posterior thalamus. On postnatal day 7, there is a sudden increase in the number of layer 6 projections to the thalamus. Surprisingly, these layer 6 cells are precisely topographically mapped with colabeled thalamic afferents on their first appearance. Subplate cells constitute a very small component of the corticothalamic projection at all ages. Double injections of DiI and DiA show that the corticofugal and thalamocortical pathways are physically separate during development. Corticofugal axons travel deep in the intermediate zone to the thalamic axons and are separate through much of the internal capsule. Their tangential distribution is also distinct. The early appearance of the thalamocortical pathway is consistent with an organizational role in the specification of some features of cortical cytoarchitecture. The specific initial projection of thalamocortical axons strongly suggests the recognition of particular cortical regions. The physical separation of these two pathways limits the possibility for exchange of information between these systems except at their respective targets. PMID- 8408773 TI - Retinotopic organization of the primary visual cortex of flying foxes (Pteropus poliocephalus and Pteropus scapulatus). AB - The representation of the visual field in the occipital cortex was studied by multiunit recordings in seven flying foxes (Pteropus spp.), anesthetized with thiopentone/N2O and immobilized with pancuronium bromide. On the basis of its visuotopic organization and architecture, the primary visual area (V1) was distinguished from neighboring areas. Area V1 occupies the dorsal surface of the occipital pole, as well as most of the tentorial surface of the cortex, the posterior third of the mesial surface of the brain, and the upper bank of the posterior portion of the splenial sulcus. In each hemisphere, it contains a precise, visuotopically organized representation of the entire extent of the contralateral visual hemifield. The representation of the vertical meridian, together with 8-15 degrees of ipsilateral hemifield, forms the anterior border of V1 with other visually responsive areas. The representation of the horizontal meridian runs anterolateral to posteromedial, dividing V1 so that the lower visual quadrant is represented medially, and the upper quadrant laterally. The total surface area of V1 is about 140 mm2 for P. poliocephalus, and 110 mm2 for P. scapulatus. The representation of the central visual field is greatly magnified relative to that of the periphery. The cortical magnification factor decreases with increasing eccentricity, following a negative power function. Conversely, receptive field sizes increase markedly with increasing eccentricity, and therefore the point-image size is approximately constant throughout V1. The emphasis in the representation of the area centralis in V1 is much larger than that expected on the basis of ganglion cell counts in flat-mounted retinas. Thus, a larger degree of convergence occurs at the peripheral representations in the retino-geniculo-cortical pathway, in comparison with the central representations. The marked emphasis in the representation of central vision, the wide extent of the binocular field of vision, and the relatively large surface area of V1 reflect the importance of vision in megachiropterans. PMID- 8408774 TI - Subcortical connections of inferior temporal areas TE and TEO in macaque monkeys. AB - To investigate the subcortical connections of inferior temporal cortex, we injected its anterior and posterior portions (Bonin and Bailey's cytoarchitectonic areas TE and TEO, respectively) in 6 rhesus monkeys with retrograde and anterograde tracers. The results indicate that both areas TE and TEO receive nonreciprocal inputs from several thalamic nuclei, including paracentralis, ventralis anterior, centralis, and limitans, and that TE also receives input from reuniens. Additional nonreciprocal inputs to both areas arise from the hypothalamus, basal nucleus of Meynert, dorsal and median raphe, locus coeruleus, and reticular formation. TE and TEO are reciprocally connected with the lateral, medial, and inferior nuclei of the pulvinar and with the ventral portion of the claustrum. The main subcortical nonreciprocal output from TE and TEO is to the striatum and from TEO to the superior colliculus. TE also sends a very limited projection to nucleus medialis dorsalis magnocellularis of the thalamus. Although the connections of areas TE and TEO are overlapping in most subcortical structures, they are partially segregated in the pulvinar, the reticular nucleus of the thalamus, and the striatum. Specifically, relative to those of TE, the projections of TEO are located more laterally in the medial, lateral, and inferior nuclei of the pulvinar, more ventrally in the reticular nucleus, and more caudally in both the ventral putamen and tail and head of the caudate nucleus. PMID- 8408775 TI - Heterogeneity of rat corticospinal neurons. AB - In order to examine the degree of diversity within a population of cortical projection neurons, rat corticospinal cells were retrogradely labeled in vivo by injecting rhodamine-tagged microspheres into the cervical spinal cord, and subsequently studied electrophysiologically and anatomically in neocortical slices maintained in vitro, by use of standard current clamp techniques and a double-labeling protocol (Tseng et al., J. Neurosci. Meth. 37:121-131, 1991). Three different subgroups were distinguished on the basis of their spiking behavior: (1) Adapting cells had a marked fast (50 ms) and slow phase (200 ms) of spike frequency adaptation; (2) regular spiking (RS) cells had only a period of fast adaptation; (3) some regular spiking neurons had prominent depolarizing afterpotentials (DAPs) and could generate bursts of spikes, often in repetitive fashion (RSDAP cells). Subgroups of RSDAP cells had different patterns of burst responses to depolarizing current pulses, suggesting differences in the types and/or sites of underlying ionic conductances. Adapting cells had a slightly higher membrane input resistance and more prominent slow hyperpolarizing afterpotentials than RS and RSDAP neurons; however, the activation of presumed anomalous rectifier current by intracellular hyperpolarizations was less prominent in adapting neurons. Orthodromic stimulation in layer I evoked presumed excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs)in all three types of cells, but prominent short-latency IPSPs were found in a higher percentage of adapting neurons. The morphology of electrophysiologically characterized corticospinal neurons was studied following intracellular injection of biocytin. All three spiking types were typical layer V pyramids with apical dendrites reaching layer 1, basal dendrites in infragranular layers, and deep directed axons that had a moderate density of local collaterals in lower cortical layers. The profuseness of dendrites, examined by Sholl's analysis of two dimensional, camera lucida-reconstructed neurons was comparable in the three neuronal subgroups, although a smaller somatic area and more slender apical dendritic trunk were found in adapting neurons. Our results suggest that corticospinal cells in rats are a heterogeneous population of projection neurons with respect to their spiking behavior, membrane properties, synaptic connections, and, to a lesser extent, their morphology. This diversity revealed in vitro adds new complexity to the classification of corticospinal neurons. PMID- 8408776 TI - Early events following challenge of rabbits with Trypanosoma evansi and T. evansi components. AB - The intradermal injection of Trypanosoma evansi or T. evansi components into rabbits evoked trypanosome-specific responses in the skin. The strongest responses, which were those against the parasite surface-associated components, had the characteristics of an immediate type hypersensitivity reaction, followed by a delayed type. The responses were greater in rabbits from which infections had been cleared by chemotherapy than in animals with patient infections. These findings suggest that variant surface glycoprotein (VSG)-specific antibody activity and immunosuppression are effective in the skin and influence the outcome of infection with T. evansi in previously infected animals. PMID- 8408777 TI - Incidence of tubulostromal adenoma of the ovary in aged germ cell-deficient mice. AB - Female mice homozygous for the germ cell-deficient (gcd) mutation enter reproductive senescence prematurely due to a dearth of germ cells arising in embryonic development. The ovaries of young gcd/gcd animals are atrophic, composed of little more than stromal cells in a connective tissue matrix. By one year of age, 56 per cent of homozygotes have developed tubulostromal adenoma of the ovary while 100 per cent wild-type and heterozygous littermates are phenotypically normal. Since these animals develop ovarian tumours more frequently as a consequence of a single autosomal recessive mutation, they will be useful models for the study of ovarian neoplasia. PMID- 8408778 TI - Neurotropism of mouse-adapted haemagglutinating encephalomyelitis virus. AB - The propagation of a mouse-adapted strain (67N) of haemagglutinating encephalomyelitis virus in infected mice and murine cells was examined by viral re-isolation and immunostaining. Viral propagation was strictly limited to the neurons and to an established line of neuroblastoma cells in in-vivo and in-vitro experiments. These results provide adequate evidence that this virus is neurotropic. PMID- 8408779 TI - Relationship between onset of puberty and establishment of persistent infection with equine arteritis virus in the experimentally infected colt. AB - The relationship between stage of reproductive tract maturity and susceptibility to the experimental establishment of persistent infection with equine arteritis virus (EAV) was investigated in 21 prepubertal and 15 peripubertal colts. Five of six prepubertal colts inoculated intranasally remained infected in the reproductive tract from post-challenge day 28 to 93 and two of six from post challenge day 120 to 180. No virus was detected in five of these animals killed on post-challenge day 210. Each of two peripubertal colts remained infected in the reproductive tract at post-challenge day 60 and one of nine was found to be persistently infected with EAV 15 months after challenge. These findings confirm that the virus can replicate in the reproductive tract of a significant proportion of colts for a variable period of time after clinical recovery in the absence of circulating concentrations of testosterone equivalent to those found in sexually mature stallions. Long-term persistent infection with EAV does not appear to occur in colts exposed to the virus before the onset of peripubertal development. We suggest that colts should be vaccinated at approximately 6 months of age, before peripubertal development but after the disappearance of maternally acquired antibodies. PMID- 8408780 TI - Histoplasmosis in horses. AB - Histoplasmosis was diagnosed in nine horses during 1986-1990. The infection with Histoplasma capsulatum caused granulomatous placentitis and abortion in one mare in the 7th month of gestation and three mares in the 10th month. Four newborn foals died from severe granulomatous pneumonia within a few days of birth; and a weanling thoroughbred developed granulomatous pneumonia and lymphadenitis at 5 months of age. PMID- 8408781 TI - Intestinal lesions in experimental phocine distemper: light microscopy, immunohistochemistry and electron microscopy. AB - The involvement of the intestinal mucosa and of the gut-associated lymphoid tissue in phocine distemper was studied in six severely diseased harbour seals 11 to 16 days after experimental infection. Five seals exhibited a mild or moderate enteritis in the small or large intestine. In all the seals, a moderate to severe depletion of submucosal lymphoid follicles was found. Likewise, antigen of phocine distemper virus (PDV) was demonstrated immunohistochemically in the intestinal wall of all the seals. Most antigen was found in the submucosal lymphoid follicles, followed by the crypt epithelium and follicle-associated epithelium (FAE). Ultrastructurally, intracytoplasmic tubular structures were detected in the FAE and interpreted as morbilliviral nucleocapsids. The results indicate a direct cytopathogenic effect of PDV on intestinal lymphoid and epithelial cells and suggest an important role of the intestinal tract in phocine distemper and, by analogy, in other morbillivirus infections as a regular site of virus replication, virus shedding and immunosuppression. PMID- 8408782 TI - Nasal epithelial changes induced in piglets by acetic acid and by Bordetella bronchiseptica. AB - Research on atrophic rhinitis of pigs has shown that both Bordetella bronchiseptica infection and experimental treatment with acetic acid predispose the nasal mucosa to colonization with Pasteurella multocida. Gnotobiotic piglets aged 3 days were dosed intranasally with either B. bronchiseptica (n = 6) or acetic acid 1 per cent (n = 10) and killed at intervals up to the 4th day after treatment. Samples of the ventral turbinates were examined by light microscopy and scanning and transmission electron microscopy. Within 12 h acetic acid induced loss of cilia, oedema, focal cell exfoliations, mitochondrial swelling and inflammatory cell infiltration. Bordetella bronchiseptica induced only a limited oedema and loss of cilia. Colonization of cilia by the bacteria was observed 96 h after infection. We conclude that, although acetic acid and B. bronchiseptica do not induce the same modifications of the nasal respiratory epithelium, their action causes stagnation of nasal mucus, which results in a nasal environment favourable to colonization by Pasteurella multocida. PMID- 8408783 TI - Multiple hepatic cavernous lymphangioma in an aged male cat. AB - A 10-year-old male Domestic Longhair cat with weight loss, enlarged left testicle, icterus and mild liver enlargement showed, at necropsy, two white, raised, firm, circumscribed masses in the liver and striking enlargement of the left testicle. The histological features of the liver masses were consistent with lymphangioma. Severe, diffuse, coalescing to multifocal, subacute, necrotizing orchitis of the left testicle also was diagnosed. A literature review and examination of the epidemiology of lymphangioma in small domestic animals demonstrated the rarity of this tumour in cats. PMID- 8408784 TI - Subclinical lumbar polyradiculopathy, polyneuritis and ganglionitis in aged wild and exotic mammalians. AB - Subclinical lumbar polyradiculopathy was present in the intradural dorsal and ventral nerve rootlets of 19 aged individuals of the following wild and exotic mammalian species: woodrat, raccoon, mink, lynx, reindeer, red deer, musk ox, scimitar-horned oryx, Arabian oryx, hybrid waterbuck, Persian onager, Przewalski's wild horse, Malayan sun bear, Asian elephant, East African river hippopotamus, vervet monkey and rhesus monkey. It was characterized by mild to severe multifocal ballooning of myelin sheaths. Occasionally, ballooned myelin sheaths contained thin strands of myelin and macrophages surrounding distorted axons. Additionally, a mild incidental lymphocytic polyneuritis was present in intradural nerve rootlets of the Malayan sun bear, and a moderate lymphocytic spinal ganglionitis in the East African river hippopotamus. PMID- 8408785 TI - Ventral meningomyelocoele in a filly. PMID- 8408786 TI - Generalized intravascular proliferation in two cats: endotheliosis or intravascular pseudoangiosarcoma? AB - Two cats had unusual occlusive vascular endothelial proliferations in several organs. The newly formed cells were strictly intraluminal and of endothelial origin, as shown by positive immunohistochemical staining with factor VIII related antigen. PMID- 8408787 TI - Nonvenereal treponematoses: yaws, endemic syphilis, and pinta. AB - The nonvenereal treponematoses--yaws, endemic syphilis, and pinta--constitute a major health concern for many third world countries. These diseases are caused by an organism that is morphologically and antigenically identical to the causative agent of venereal syphilis, Treponema pallidum. Nonvenereal treponematoses differ significantly in their modes of transmission, epidemiology, and clinical presentation from venereal syphilis. Like venereal syphilis, they have a chronic relapsing course and have prominent cutaneous manifestations. Recently, several cases of imported yaws and endemic syphilis have been described in Europe. With the escalating U.S. military presence in many remote areas of the world and ever increasing world-wide travel, the diagnosis of the nonvenereal treponematoses must be considered in appropriate clinical and historical situations. PMID- 8408788 TI - Complement polymorphism in herpes gestationis: association with C4 null allele. AB - BACKGROUND: Herpes gestationis (HG) is a rare, pregnancy-related skin disease characterized by the production of an autoantibody to a component of the hemidesmosome. It is associated with the class II antigens HLA-DR3 and HLA-DR4, but its potential association with the "class III antigens" C2, C4, and factor B has not previously been studied. OBJECTIVE: Our purpose was to study complement polymorphism in HG. METHODS: Using electrophoresis and immunofixation techniques, we determined the allele frequencies of C4A, C4B, C3, and factor B in 42 patients with a history of HG. RESULTS: Ninety percent of patients carried a C4 null allele (C4*QO). No statistically significant association with C3 or factor B alleles was seen. CONCLUSION: HG is associated with the presence of a C4*QO. Whether the C4*QO is the primary genetic association, or whether the C4*QO is related to its linkage disequilibrium with DR3 and DR4 has yet to be determined. PMID- 8408789 TI - Posttransplant cutaneous lymphoma. AB - BACKGROUND: Posttransplant lymphoma is closely associated with Epstein-Barr virus (EBV) infection and appears to have a predilection for extranodal sites. We describe four cases of primary cutaneous posttransplant lymphoma. OBJECTIVE: Our aim was to determine cell lineage and any possible association with EBV in each case of cutaneous lymphoma. METHODS: Tumor tissue was examined by light microscopy, immunohistochemistry, nonisotopic in situ hybridization and polymerase chain reaction. RESULTS: The data were consistent with a diagnosis of EBV-associated cutaneous B-cell lymphoma in three cases and cutaneous T-cell lymphoma not associated with EBV in one case. No patient with B-cell lymphoma had extracutaneous involvement during a mean follow-up of 3.9 years. The patient with cutaneous T-cell lymphoma died of cerebral involvement 9 months after initial presentation. CONCLUSION: These data suggest a possible role for EBV infection in the origin of cutaneous B-cell lymphoma in immunosuppressed patients. PMID- 8408790 TI - Prediction of histologic melanocytic dysplasia from clinical observation. AB - BACKGROUND: The dysplastic melanocytic nevus (DMN) is a putative precursor for malignant melanoma in persons with a family history of melanoma and multiple large atypical moles. Furthermore, the concept that DMN confers increased risk for melanoma has been extended to those without a family history of melanoma. Some investigators have characterized DMN in terms of size, surface topography, and irregularity of border and color and have provided definitive statements as to major and minor clinical "criteria" for DMN. OBJECTIVE: In this study we conducted a quantitative test of previously described clinical characteristics in patients diagnosed with melanoma but without a family history of melanoma in a first-degree relative (N = 153). METHODS: Two of us examined each patient independently, counting the total number of nevi, deciding on the most clinically atypical nevus, and recording quantitatively the size, surface, border, and color characteristics. The most atypical nevus was removed from each patient and diagnosed histologically without knowledge of the clinical examiner's description. RESULTS: Of the 153 nevi so examined, 91 (59.5%) were judged to be clinically atypical and 23 nevi (15%) were classified as histologically dysplastic. We then analyzed the ability of each clinical feature to predict histologic melanocytic dysplasia. A multivariate analysis was conducted with the blindly scored clinical features as independent variables. Independent variables included total number of nevi and freckles as well as the features of the particular nevus biopsied (including longest diameter, macular component, irregular border, ill-defined border, haphazard coloration). Of the many independent clinical variables, only total number of nevi and macular component were useful in the final multivariate prediction. Of 21 nevi with a macular component removed from persons with more than 24 total body nevi, 7 were histologically dysplastic, representing a positive predictive value of 33.3% (95% exact confidence limits 14.6% to 57.0%). Of the 38 nevi without a macular component to their most atypical nevus removed from patients with 12 or fewer total body nevi, 37 failed to meet histologic criteria for DMN, representing a negative predictive value of 97% (95% exact confidence limits 86.2% to 99.9%). CONCLUSION: We conclude that in nonfamilial melanoma, clinical criteria suggesting DMN when present will often fail to display histologic melanocytic dysplasia. In contrast, absence of a macular component in melanocytic nevi in a person with fewer than 13 total body nevi will accurately predict the absence of melanocytic dysplasia on histologic examination. PMID- 8408791 TI - An evaluation of oral ulcers in patients with AIDS and AIDS-related complex. AB - BACKGROUND: Patients with HIV infection can have recurrent and persistent oral ulcers, not attributable to known infectious agents. OBJECTIVE: Our aim was to evaluate prospectively oral ulcers in patients with HIV infection to determine whether an etiologic agent could be identified. METHODS: Sixteen patients with HIV infection who had oral ulcers not attributable to known causes had culture of the base and a biopsy specimen taken from the ulcer. Cultures were obtained for herpes simplex and varicella-zoster viruses, mycobacteria, and fungi. By polymerase chain reaction (PCR) analysis with primer/probe sets for herpes simplex viruses 1 and 2, varicella-zoster virus, cytomegalovirus, human papillomavirus, and Mycobacterium tuberculosis, each biopsy specimen was analyzed for the presence of DNA from these organisms. Specimens were also evaluated histologically. RESULTS: Histoplasmosis was detected histologically in one biopsy specimen, candidiasis in a second, and herpetic changes in a third. Viral cultures were positive for herpes simplex virus 1 in four cases and herpes simplex virus 2 in one case. PCR analysis detected DNA for herpes simplex virus 1 in one case and herpes simplex virus 2 in another; DNA from other pathogens was not identified. In the remaining eight patients, hematoxylin-and-eosin staining revealed eosinophilic ulcers in five cases and nonspecific changes in three cases. CONCLUSION: The etiologic agent of recurrent or persistent oral ulcers in patients with AIDS and AIDS-related complex was not identified in 50% of patients. PCR analysis was not useful. Herpes simplex virus or other pathogens were not detected in ulcers containing numerous eosinophils. PMID- 8408792 TI - Psychological effects of androgenetic alopecia on women: comparisons with balding men and with female control subjects. AB - BACKGROUND: Several studies have examined the psychological impact of androgenetic alopecia on men but scientific evidence is absent regarding its effects on women. OBJECTIVE: Our purpose was to determine the psychosocial sequelae of androgenetic alopecia in women and, comparatively, in men. METHODS: Subjects were newly referred patients with androgenetic alopecia (96 women and 60 men) and 56 female control patients. Subjects completed standardized questionnaires to assess their psychological reactions to their respective conditions and to measure body image, personality, and adjustment. RESULTS: Androgenetic alopecia clearly was a stressful experience for both sexes, but substantially more distressing for women. Relative to control subjects, women with androgenetic alopecia possessed a more negative body image and a pattern of less adaptive functioning. Specific correlates of the adversity of patients' hair loss experiences were identified. CONCLUSION: The results confirm the psychologically detrimental effects of androgenetic alopecia, especially on women. The implications for patient care are discussed. PMID- 8408793 TI - Comparison of the local and systemic side effects of methylprednisolone aceponate and mometasone furoate applied as ointments with equal antiinflammatory activity. AB - BACKGROUND: The therapy for skin diseases with topical glucocorticoids is limited by their local and systemic side effects. A glucocorticoid with an improved benefit-to-risk ratio is desirable. OBJECTIVE: A new topical corticoid, methylprednisolone aceponate (MPA) 0.1% ointment, was compared with the same formulation of mometasone furoate. METHODS: The two ointments were compared with respect to suppression of UVB light-induced erythema (n = 20) and with respect to atrophogenicity and appearance of telangiectasia (n = 20) in two double-blind trials with intraindividual comparisons in healthy volunteers. In a third trial, serum cortisol levels were measured in volunteers receiving extensive (60% of body surface) cutaneous application of MPA (n = 10) or mometasone furoate (n = 11). RESULTS: MPA and mometasone furoate were equally effective in suppressing UVB light-induced erythema. Atrophogenicity, as well as the incidence and severity of telangiectasia, were significantly more pronounced with mometasone furoate than with MPA. Both ointments decreased serum cortisol levels and did not differ significantly in this respect. However, the incidence of serum cortisol level suppression was higher in the mometasone furoate group than in the MPA group. CONCLUSION: MPA ointment has equal antiinflammatory activity and similar cortisol suppression but significantly fewer local side effects than mometasone furoate. PMID- 8408794 TI - Statistical evaluation of epiluminescence microscopy criteria for melanocytic pigmented skin lesions. AB - BACKGROUND: Epiluminescence microscopy (ELM) is a noninvasive technique by which the clinical diagnosis of pigmented skin lesions (PSL) can be improved. Many ELM criteria have been described, but their significance in the differential diagnosis of PSL has not yet been established. OBJECTIVE: The purpose of this study was to determine the value of ELM criteria in the differential diagnosis of PSL. METHODS: Two hundred one melanocytic PSL (61 common nevi, 60 dysplastic nevi, and 80 melanomas) were investigated with ELM for the presence of certain ELM criteria; their significance was determined by calculating the odds ratios. RESULTS: Individual ELM criteria have different weights of significance in the differential diagnosis of melanocytic PSL. Selected patterns of ELM criteria adjusted to the distinct types of PSL considerably improve the diagnostic accuracy of melanocytic PSL. CONCLUSION: The prevalence of certain distinct ELM criteria in a given melanocytic PSL has statistical value in differential diagnosis. PMID- 8408795 TI - Management of lentigo maligna and lentigo maligna melanoma with paraffin-embedded tangential sections: utility of immunoperoxidase staining and supplemental vertical sections. AB - BACKGROUND: The use of frozen sections in the management of lentigo maligna and lentigo maligna melanoma has been the focus of some controversy. OBJECTIVE: Our purpose was to utilize paraffin-embedded tangential sections in the management of two cases of lentigo maligna and three cases of lentigo maligna melanoma. METHODS: A modification of Mohs micrographic surgery using rush paraffin-embedded sections with adjunctive immunoperoxidase staining (HMB-45) and supplemental vertical sections was employed. RESULTS: This method resulted in enhanced histologic evaluation of section margins and did not compromise the diagnosis of the primary invasive melanoma or Breslow measurements. CONCLUSION: Mohs micrographic surgery modified by the use of rush paraffin-embedded sections allows adjunctive immunoperoxidase staining and supplemental vertical sections that may be helpful in the management of lentigo maligna and lentigo maligna melanoma. PMID- 8408796 TI - Dermatologic therapy: 1992. AB - This article reviews therapeutic studies reported in the English-language literature during 1992. Readers should review the original article in full before attempting any experimental or controversial therapy. PMID- 8408797 TI - Does pregnancy influence the prognosis of malignant melanoma? AB - The effect of pregnancy on the clinical course of malignant melanoma (MM) is unclear. Early clinical and laboratory evidence suggested a relation between hormones and MM and subsequently between pregnancy and MM. We reviewed the literature on MM and pregnancy to address three questions: What is the effect on prognosis if an MM is diagnosed during pregnancy? What is the effect of previous pregnancies on the prognosis of MM? What effect does a subsequent pregnancy have on the prognosis of MM? On the basis of a limited number of controlled studies, it does not appear that being pregnant before, after, or at the time of diagnosis of stage I MM influences the 5-year survival rate. However, caution in interpreting these data must be taken because it is possible that the duration of follow-up and size of the study populations are not sufficient to observe a true effect. PMID- 8408798 TI - Clinical pearl: proximal white subungual onychomycosis in AIDS. PMID- 8408799 TI - Surgical pearl: a user-friendly dressing. PMID- 8408800 TI - Lymphocutaneous nocardiosis caused by Nocardia caviae: the first case report from Asia. PMID- 8408801 TI - Clear cell variant of mucoepidermoid carcinoma of the skin. PMID- 8408802 TI - Extensive human papillomavirus-related disease (bowenoid papulosis, Bowen's disease, and squamous cell carcinoma) in a patient with hairy cell leukemia: clinical and immunologic evaluation after an interferon alfa trial. PMID- 8408803 TI - Efficacy of a 1-week, once-daily regimen of terbinafine 1% cream in the treatment of tinea cruris and tinea corporis. PMID- 8408804 TI - Annular bullous eruption in a patient with the CREST syndrome, primary biliary cirrhosis, and Sjogren's syndrome. PMID- 8408805 TI - Ascher syndrome: a mimicker of acquired angioedema. PMID- 8408806 TI - Symmetric pyogenic granuloma. PMID- 8408807 TI - Spontaneous regression of Merkel cell (neuroendocrine) carcinoma of the skin. PMID- 8408808 TI - Metastatic adenocarcinoma of the scalp mimicking a kerion. PMID- 8408809 TI - Cutaneous aspergillosis complicating pyoderma gangrenosum. PMID- 8408810 TI - Sun between 3 PM and 4 PM can burn! PMID- 8408811 TI - Annular bullous pemphigoid in a child. PMID- 8408812 TI - Adult Kawasaki syndrome associated with HIV infection and anticardiolipin antibodies. PMID- 8408813 TI - Are antibodies to neutrophilic cytoplasmic antigens (ANCA) a serologic marker for Sweet's syndrome? PMID- 8408814 TI - Treatment of reticulate acropigmentation of Kitamura with azelaic acid. PMID- 8408815 TI - HIV and alopecia universalis. PMID- 8408816 TI - Spitz nevi. PMID- 8408817 TI - Prolidase deficiency: a multisystemic hereditary disorder. AB - Prolidase deficiency is a rare hereditary disorder with a wide spectrum of clinical manifestations including skin ulcers, eczematous eruptions, characteristic facies, mental retardation, splenomegaly, and susceptibility to infections. We report two new cases of prolidase deficiency. Our patients had the typical manifestations of prolidase deficiency. One also had lupus erythematosus. Prolidase activity was either normal or half-normal in all family members. The skin disease in our patients did not respond to topical glycine/proline ointment or to oral vitamin C. PMID- 8408818 TI - Granulomatous cheilitis in two children with sarcoidosis. AB - Sarcoidosis is rare in children, who usually have only asymptomatic pulmonary involvement. We report granulomatous cheilitis in two children with sarcoidosis. Both children had systemic sarcoidosis with lymph node and pulmonary involvement. PMID- 8408819 TI - Paraneoplastic pemphigus presenting as an unusual bullous eruption. AB - We describe a patient with a widespread annular bullous eruption on the trunk and extremities and involvement of the conjunctivae, oral mucous membranes, and esophagus. Findings of a biopsy specimen of an intraabdominal mass revealed nodular and diffuse well-differentiated lymphoma. Histologic examination of a skin lesion showed acantholysis with bulla formation and keratinocyte necrosis. Direct immunofluorescence testing of a skin lesion showed intercellular deposition of IgG. The patient's serum contained autoantibodies with broad specificity for skin and other epithelia and identified the characteristic bands of paraneoplastic pemphigus at 250, 230, 210, and 190 kd on immunoprecipitation. The patient died despite therapy. PMID- 8408820 TI - Terminal hepatic failure in erythropoietic protoporphyria. AB - Erythropoietic protoporphyria is an inherited disorder characterized biochemically by a deficiency of ferrochelatase, the enzyme that catalyzes the incorporation of ferrous iron into protoporphyrin to form heme. We describe a patient who illustrates the unpredictability of the course of liver disease in erythropoietic protoporphyria. She remained stable for several years after her first evidence of liver function abnormalities. Then, in a period of weeks, hepatic failure developed and she died. Findings of serial liver biopsy specimens showed extensive hepatocellular degeneration and inflammation that appeared in a 10-day period. The factors that cause this rapid deterioration in hepatic function remain unknown. Reported cases of fatal hepatic failure in patients with erythropoietic protoporphyria are reviewed. PMID- 8408821 TI - Localized argyria with chrysiasis caused by implanted acupuncture needles. Distribution and chemical forms of silver and gold in cutaneous tissue by electron microscopy and x-ray microanalysis. AB - A case of localized argyria with chrysiasis caused by implanted acupuncture needles in a 41-year-old Japanese woman was studied by electron microscopy and x ray microanalysis. Large amounts of silver granules with selenium and sulfur were detected around eccrine secretory cells in much greater amounts than around ductal cells. Many granules were also observed along the outer edge of the basement membrane but never within cells or intercellular spaces. The granules were also present around blood vessels, lymphatics and nerve fibers, and in elastic fibers. Small numbers of gold fragments were also seen, mostly within macrophages. These results suggest that silver deposits extracellularly as selenide and sulfide, whereas free gold is found intracellularly. PMID- 8408822 TI - Unilateral emboli in a patient with thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura is a rare disease most commonly associated with microangiopathic hemolytic anemia, thrombocytopenia, fever, neurologic disorders, and renal dysfunction. We describe a patient with a history of thrombotic thrombocytopenic purpura that had been quiescent for 4 months; he had a 3-week history of painful purpuric lesions on the left hand only. He also had mottling and a livedoid purpura of the distal fingertips, splinter hemorrhages of the left fingernails, and a decreased radial pulse. Findings of a biopsy specimen revealed multiple capillary and small vessel thromboses. Contrast aortography demonstrated a pseudoaneurysm of the proximal descending thoracic aorta with stenosis of the left subclavian artery at its origin and an associated thrombus. PMID- 8408823 TI - Microcystic adnexal carcinoma. AB - Microcystic adnexal carcinoma is a rare cutaneous neoplasm characterized by invasive, relentless, and destructive local growth. The incidence of perineural invasion and tumor recurrence is high. We report two cases of microcystic adnexal carcinoma with typical clinical features. The use of formalin-fixed, paraffin embedded horizontal Mohs sections is described in one case. We also review the current concepts and management of microcystic adnexal carcinoma. PMID- 8408824 TI - Acquired cutis laxa with dermatitis herpetiformis and sarcoidosis. AB - Cutis laxa is a rare condition characterized by loss of elastic tissue. We describe a patient with dermatitis herpetiformis and pulmonary and articular sarcoidosis in whom acquired cutis laxa developed. We believe this association is unique. PMID- 8408825 TI - La (SS-B)-positive neonatal lupus erythematosus: report of a case with unusual features. AB - Neonatal lupus erythematosus is most often associated with autoantibodies against Ro and La antigens. Rarely, neonatal lupus erythematosus occurs in the absence of Ro antibody. We present a case of La antibody-positive neonatal lupus erythematosus with unusual features. PMID- 8408826 TI - Cutaneous necrosis resulting from protein S deficiency and increased antiphospholipid antibody in a patient with systemic lupus erythematosus. AB - A 23-year-old woman with widespread cutaneous necrosis resulting from protein S deficiency and increased antiphospholipid antibody in the setting of systemic lupus erythematosus is described. Our report illustrates the complex nature of evaluation of skin lesions in patients with active systemic lupus erythematosus. PMID- 8408827 TI - Granuloma annulare perforans in herpes zoster scars. AB - Granuloma annulare perforans limited to a thoracic dermatome that was previously involved by herpes zoster occurred in a 51-year-old woman who also had Lennert's lymphoma. Of the various local granulomatous infiltrates described after herpes zoster, granuloma annulare perforans is unique, although ordinary granuloma annulare has been described in a few patients. A high incidence of specific and nonspecific reaction patterns in herpes zoster scars has been described in patients with malignant lymphoma. In contrast to previous patients, all of whom had chronic lymphatic leukemia, our patient had Lennert's lymphoma. PMID- 8408828 TI - In utero acute graft-versus-host disease in a neonate with severe combined immunodeficiency. AB - We describe a male neonate with severe combined immunodeficiency who at birth had acute graft-versus-host disease (GVHD) as a result of maternal-fetal transfusion during pregnancy. Several clinical signs helped establish this diagnosis. Findings of a skin biopsy specimen confirmed the diagnosis of acute GVHD. Immunologic evaluation disclosed an absence of T and B lymphocytes. Acute GVHD in severe combined immunodeficiency most often occurs during the first weeks of life; intrauterine occurrence is unusual. PMID- 8408829 TI - Reflex sympathetic dystrophy with prominent involvement of the nail apparatus. AB - After closed hand trauma, a 17-year-old boy had acute inflammatory changes that resembled bacterial whitlows of the third and fourth right fingers. Clearing of the inflammatory changes was followed by the development of cyanosis, hyperhidrosis, and roentgenographic evidence of patchy osteoporosis in the involved extremity. Findings of a biopsy specimen revealed that the inflammatory lesions in the proximal nail folds were caused by proliferation of capillary vessels embedded in edematous loose connective tissue. This is the first report of cutaneous histopathologic findings in the first stage of reflex sympathetic dystrophy, although similar features have been described in synovial and bone biopsy specimens of patients with reflex sympathetic dystrophy. PMID- 8408830 TI - Unusual aspects of juvenile xanthogranuloma. AB - We describe three unusual features of juvenile xanthogranuloma that were observed in three different children. We also describe the mixed and clustered forms of juvenile xanthogranuloma and a giant juvenile xanthogranuloma of the nose. PMID- 8408831 TI - Primary CD8+ lymphoepithelioid lymphoma of the skin. AB - We describe an 82-year-old man with CD8+ lymphoepithelioid lymphoma localized to the skin. The skin lesion had been present for 18 years. Histologically there was a dense infiltrate that consisted of atypical lymphoid cells and large epithelioid cells, thereby resembling the so-called Lennert's lymphoma. Local radiotherapy proved effective in reducing the size of the tumor. To the best of our knowledge, this is the first report of primary lymphoepithelioid lymphoma of the skin. PMID- 8408832 TI - Bullous pyoderma gangrenosum. AB - Pyoderma gangrenosum is a necrotizing, ulcerative process commonly associated with inflammatory bowel disease but also occurring in several other systemic illnesses. When associated with myelodysplastic syndromes, its clinical appearance is often atypical. We describe a patient with severe, refractory bullous pyoderma gangrenosum. At autopsy cardiac involvement was demonstrated, a finding not previously reported. Patients with this unusual variant of pyoderma gangrenosum have a grave prognosis. PMID- 8408833 TI - Antigen identification in drug-induced bullous pemphigoid. AB - Immunobullous diseases usually develop spontaneously, but drug-induced bullous disease develops in a small subgroup of patients. We examined a patient in whom bullous pemphigoid developed after she received enalapril for treatment of hypertension. IgG antibody directed against a 230 kd antigen was identified. The eluted IgG autoantibody was shown to bind to the basement membrane zone on split skin. This study demonstrates that drug-induced bullous pemphigoid autoantibody in this patient was directed against the same antigen as the spontaneous bullous pemphigoid antigen. PMID- 8408834 TI - Xeroderma pigmentosum-Cockayne syndrome complex in two patients: absence of skin tumors despite severe deficiency of DNA excision repair. AB - Two brothers had a complex combination of two DNA repair disorders: Cockayne syndrome and xeroderma pigmentosum. This rare combination has previously been observed in only two other patients. The clinical signs shared by these two brothers and the two other previously described patients include severe sun sensitivity, freckling, diminished stature, hearing and movement impairment, and neurologic degeneration. Although defective UV-induced unscheduled DNA synthesis has been demonstrated (5% of normal), no skin cancers have appeared in these 38- and 41-year-old brothers, whereas skin cancers developed at a relatively early age in the two previously described patients who also had defective UV-induced unscheduled DNA synthesis. PMID- 8408835 TI - Kaposi's sarcoma with visceral involvement after intraarticular and epidural injections of corticosteroids. AB - Kaposi's sarcoma has been reported in patients receiving immunosuppressive therapy, most of whom are organ transplant recipients. The development of Kaposi's sarcoma after treatment with corticosteroids has been reported in only 38 patients who have not had acquired immunodeficiency syndrome or undergone organ transplantation. Cutaneous Kaposi's sarcoma developed 2 months after intraarticular steroid injections in a man with ulnar nerve entrapment. The lesions regressed spontaneously after 3 months but reappeared with visceral involvement 18 months later, shortly after initiation of a course of epidural steroid injections for treatment of low back pain. The cutaneous lesions and some visceral lesions rapidly regressed after cessation of treatment. PMID- 8408836 TI - Maffucci's syndrome: two case reports with a literature review. AB - Sixty-three cases of Maffucci's syndrome in the English literature plus two additional cases of our own are reviewed. This syndrome is nonhereditary and is characterized by multiple enchondromas and hemangiomas. It occurs in all races with no sex predominance. The enchondromas and hemangiomas can occur anywhere but are most common in the hands. Long bone involvement is common and leads to progressive skeletal deformity and pathologic fractures. The incidence of malignancies in patients with this syndrome is high. Chondrosarcomas are especially common and occur in 30% of the patients. PMID- 8408837 TI - Bullous pemphigoid and associated autoimmune thrombocytopenia: two case reports. AB - We describe two patients with bullous pemphigoid in whom autoimmune thrombocytopenia developed. Only one case of bullous pemphigoid associated with autoimmune thrombocytopenia has previously been reported, and in that case, autoimmune hemolytic anemia was also present (Evans' syndrome). However, a wide range of other autoimmune diseases have been described in association with bullous pemphigoid, and this literature is reviewed. PMID- 8408838 TI - Febrile ulceronecrotic Mucha-Habermann disease. AB - Febrile ulceronecrotic Mucha-Habermann disease in an 18-year-old man is reported. This disease is a severe form of pityriasis lichenoides et varioliformis acuta (PLEVA) and is characterized by the sudden onset of diffuse coalescent ulcerations associated with high fever and systemic symptoms. In the present case the disease was preceded by typical PLEVA. Histologically, a leukocytoclastic vasculitis was seen in addition to the usual features of PLEVA. Findings of laboratory studies revealed an elevated erythrocyte sedimentation rate, a high white blood cell count, and a mild increase in liver enzymes. No systemic involvement was detected. Findings of T cell receptor gene analysis in skin and peripheral blood showed no abnormality. The patient was treated with PUVA and methotrexate with a good response. We review the eight previously reported cases of febrile ulceronecrotic Mucha-Habermann disease. PMID- 8408839 TI - Laryngo-onycho-cutaneous syndrome associated with familial benign hypercalcemia. AB - A Pakistani boy had chronic ulceration of the cheeks, generalized nail dystrophy, ulceration and hypergranulation of the nail beds, conjunctiva, and vocal cords, and dental enamel hypoplasia. These features are consistent with laryngo-onycho cutaneous (LOCS) syndrome or Laryngeal and Ocular Granulation tissue in children from the Indian subContinent (LOGIC) syndrome. Both he and his elder brother had elevated levels of serum calcium consistent with familial benign hypercalcemia. Laryngo-onycho-cutaneous syndrome is a newly recognized condition and its association with familial benign hypercalcemia has not been previously reported. PMID- 8408840 TI - Fatal septicemia and bullae caused by non-01 Vibrio cholerae. AB - Bullous lesions associated with non-01 Vibrio cholerae developed in a patient with hepatic cirrhosis who had recently ingested raw oysters. He died of overwhelming sepsis despite 5 days of aggressive antibiotic therapy. Non-01 V. cholerae was isolated from blood, peritoneal fluid, and bullae. The organism produced a cytotoxic factor that destroyed Chinese hamster ovary cells. Although septicemia caused by non-01 V. cholerae is uncommon, cutaneous manifestations of this organism are even rarer. Our patient represents the first reported case of bullous lesions associated with non-01 V. cholerae septicemia. PMID- 8408841 TI - Methotrexate and reproduction in men: case report and recommendations. AB - Methotrexate has been used in the treatment of recalcitrant psoriasis for more than 35 years. We examined the significance of impaired spermatogenesis in a young man undergoing methotrexate treatment for severe psoriasis with associated arthritis. A medical geneticist was consulted and a review of the literature was performed. Genetic abnormalities that could lead to mutagenesis include chromosomal abnormalities and single-gene mutations. These aspects are considered and recommendations are made for counseling men undergoing methotrexate therapy so that risks and options can be considered. PMID- 8408842 TI - Different sensitivity to amiloride of body and tail skins of Rana catesbeiana tadpoles during metamorphosis. AB - Regional differences in potential difference and short-circuit current between the body (dorsal) and the tail skin during metamorphosis of Rana catesbeiana tadpoles were investigated. In body skin, the potential difference and the short circuit current across the skin develop in two successive steps. At stage XX, the potential difference and the short-circuit current across the body skins were amiloride-insensitive (1st step). At stage XXII, however, amiloride-sensitive potential difference and the short circuit current appeared (2nd step). By contrast, in tail skin the potential difference and the short-circuit current remained amiloride-insensitive (1st step) even at stage XXIII. Since the tail regresses after stage XXIII, the appearance of the second step could not be followed in vivo. To determine whether or not the second step can be induced in the tail, tail skin was cultured under conditions where the skin survives for a much longer period than it does in normally developing tadpoles. Such cultured tail skin generated the amiloride-sensitive potential difference and the short circuit current and cultured body skin also generated them. Therefore, development of the 2nd step in the tail skin may be delayed in vivo. To characterize the differences between body and tail skin, skins were mutually grafted between body and tail at stage XIII-XV. The body skin grafted on the tail underwent both the 1st and 2nd steps by stage XXII, whereas the tail skin grafted on the body only showed the 1st step by the same stage. These results suggest that the regional specificity of the skin is already established before the prometamorphic stage. PMID- 8408843 TI - Ca2+ transport and chemoreception in Paramecium. AB - Intracellular Ca2+ levels in Paramecium must be tightly controlled, yet little is understood about the mechanisms of control. We describe here indirect evidence that a phosphoenzyme intermediate is the calmodulin-regulated plasma membrane Ca2+ pump and that a Ca(2+)-ATPase activity in pellicles (the complex of cell body surface membranes) is the enzyme correlate of the plasma membrane pump protein. A change in Ca2+ pump activity has been implicated in the chemoresponse of paramecia to some attractant stimuli. Indirect support for this is demonstrated using mutants with different modifications of calmodulin to correlate defects in chemoresponse with altered Ca2+ homeostasis and pump activity. PMID- 8408844 TI - Water loss from eggs of domestic fowl and calcium status of hatchlings. AB - Water balance in eggs of domestic fowl was manipulated by drilling holes (each 3 mm in diameter) through the calcareous layer into the air cell on day 10 of incubation. Water loss between days 0 and 18 averaged 6 g for eggs in the control group (no hole) but increased to 8, 12, and 15 g for eggs with 1, 2, or 3 holes, respectively. Hatching success was 79-87% for eggs with 0-2 holes through the eggshell but only 43% for eggs with three holes. Live mass of hatchlings declined as the number of holes drilled in the eggshell increased, but dry mass of carcasses was unaffected by the treatments. The quantity of Ca2+, Mg2+, and phosphorus in residual yolks and yolk-free carcasses of hatchlings was not influenced by the amount of water lost from eggs. Plasma Ca2+ and Mg2+ were elevated in hatchlings from eggs with high rates of water loss. The inability to regulate plasma Ca2+ may be a negative consequence of excessive water loss and could contribute to increased mortality of embryos. PMID- 8408845 TI - Studies on the growth-stimulatory activity of pigeon milk--comparison and synergistic effects with serum. AB - Pigeon milk, a nutritive secretion from the crop of breeding pigeons, was tested (on v/v basis) for growth factor activity either separately or in combination with other growth supplements. Synthesis of DNA in confluent monolayers of quiescent Chinese hamster ovary cells was enhanced by the homogenates of pigeon milk in the presence of both fetal bovine serum and bovine serum albumin, although the response with fetal bovine serum was greater than that with bovine serum albumin. The in vitro growth stimulation by pigeon milk was also reflected in the increase in cell number. Specific activity of pigeon milk growth factor, measured against both Chinese hamster ovary cells and mouse embryo fibroblasts, was found to be higher than that of fetal calf serum, fetal bovine serum, and goat, horse, pig and human serum. The growth-stimulatory property of pigeon milk did not change in the first 5 days of its secretion. PMID- 8408846 TI - The effect of diet quality on gut anatomy in British voles (Microtinae). AB - Three species of British voles, the bank vole Clethrionomys glareolus, the field vole Microtus agrestis and the water vole Arvicola terrestris were maintained on diets of seed and plant leaf material to investigate changes in gut anatomy. C. glareolus and M. agrestis showed significant changes in most regions of the gut; they developed longer and heavier tracts when on a high-fibre diet. This response may be important in enabling these animals to withstand seasonal changes in diet quality. PMID- 8408847 TI - The multicultural workplace: preparing preceptors. AB - As we approach the 21st century, the workforce in the United States is becoming more culturally diverse. This is evident in many of our hospitals across the nation. Preceptors, who are trained as role models, communicators, and teachers, are poorly prepared to deal with the cultural diversity of nurses and their clients. The Texas Medical Center Collaborative Preceptor Program uses several methods to explore cultural differences and increase preceptors' sensitivity to people from other cultures. The course assists preceptors in identifying potential conflicts of value systems and cultural differences and enables them to identify methods of facilitating cultural adjustment. Preceptors are given the opportunity to identify their own cultural values and communication techniques and to experience cultural differences firsthand. The class includes lecture/discussion, gaming, role-play, and video. Experiential learning is used to identify cultural differences without being culturally offensive or stereotypical. PMID- 8408848 TI - Job burnout in RN-to-BSN students: relationships to life stress, time commitments, and support for returning to school. AB - Returning to school is accompanied by changes in role in the work setting and in the home setting, each of which may result in increased stress and conflict and may lead to burnout for RNs. A convenience sample of 54 students enrolled in an RN baccalaureate-completion program were surveyed to explore the relationships among life stress; the multiple time commitments of work, school, and home life; perceived support for returning to school from employer, colleagues, and family; and burnout in the work setting. Pearson's correlations indicated significant negative relationships among burnout and support from family and colleagues, and perceived control of the situation. There were no significant relationships between burnout and life stress or time commitments. PMID- 8408849 TI - Self-paced RN refresher program: the North Carolina experience. AB - The need for refresher courses is a constant, as is the lack of courses, faculty, and geographically sufficient numbers of students. This article describes an approach that North Carolina nurses developed to address the need for RN refresher programs. PMID- 8408850 TI - A qualitative analysis of factors in the work environment that influence nurses' use of knowledge gained from CE programs. AB - Continuing education (CE) programs for nurses often result in improved quality of patient care and personal and professional growth of participants. These outcomes are not achieved when nurses perceive barriers to putting knowledge from CE programs into practice. This qualitative study examined nurses' perceptions of factors in the work environment that influence the use of knowledge gained from CE programs. Fourteen themes were identified from analysis of in-depth interviews of eight RNs and observations of their work environment, a 40-bed general medical unit. Factors identified by the nurses were rated on a Likert-type scale and these factors were represented in diagrammatic form, consistent with Lewin's (1951) Field Theory analysis. An in-depth analysis of two qualitative themes is presented here. This analysis suggests three important implications for educators and reemphasizes the need for participant involvement in the planning phase of continuing education programs. PMID- 8408851 TI - Continuing education for advanced oncology nurses. AB - Continuing education (CE) opportunities for oncology nurses practicing at an advanced level are difficult to access. This article describes an ongoing educational effort to meet the needs of this target group in a midwestern state. Details of conference planning, implementation, and evaluation are included. A description of the screening process and program content is emphasized. This approach is applicable to a variety of specialty practice areas. PMID- 8408852 TI - Hospitals and the educational institution: an innovative partnership for nursing specialty education. AB - Rapid technological and economic change require that educational institutions respond efficiently and effectively to the needs of the health care industry. Nurses demand education that is accessible, affordable, and relevant, and demand recognition for educational achievement in terms of credentials and financial rewards. This article describes a partnership between British Columbia Institute of Technology (which is driven by and responds to industry) and The School of Health Science (which prepares nurses for specialty practice). The partnership permits a proactive relationship that utilizes the best qualities of both the health care and educational institutions. Although this alliance has been used extensively to prepare nurses for specialty areas, this partnership model could be implemented by a variety of institutions to meet educational needs in many health care professions. PMID- 8408853 TI - Continuing education needs of hospital-based nurses in Alabama. AB - Staff nurses (n = 403) and nurse executives (n = 82) were surveyed to determine the continuing education (CE) needs of hospital-based nurses in Alabama. Nurse executives preferred on-site, live offerings on Wednesday. Staff nurses preferred on-site, live offerings on Monday or Tuesday. Findings indicated that there was a difference in: (a) nurse executives' and staff nurses' perception of reimbursement policies, (b) nurse executives' and staff nurses' choice for days of week preferred, and (c) reimbursement policies and practices for RNs and LPNs. PMID- 8408854 TI - Copying of home study course materials: review of fair-use copying criteria. PMID- 8408855 TI - E.T., Fido, and 20 of us. PMID- 8408856 TI - Diversity development. PMID- 8408857 TI - On impostorship, cultural suicide, and other dangers: how nurses learn critical thinking. AB - Critical thinking is an important idea whose time seems to have come for nursing education. However, as a newly fashionable buzzword, it is in danger of being uncritically accepted, of being regarded as the panacea for all problems of nursing practice. This article goes behind the positive rhetoric of critical thinking to explore how this process manifests itself in the lives of nursing practitioners, nurse managers, administrators, and educators. Drawing on nurses' own stories of what happened as they challenged conventional professional or organizational assumptions and as they explored alternative perspectives on nursing practice, a picture of critical thinking "from the inside" emerges. Themes of impostorship, cultural suicide, lost innocence, roadrunning, and community emerge as defining features of the critical thinking process. It becomes clear that critical thinking is a strongly emotional as well as cognitive process, and that it carries considerable political dangers for its protagonists. PMID- 8408858 TI - Learning environment: the catalyst for work excitement. AB - A recent study investigated the relationship between learning environment and work excitement among a group of nurses. In a secondary analysis of the original survey of 268 nurses, approximately 20% were very excited about their work. The learning environment variables (availability of learning opportunities; stimulating, motivating, challenging environment; and opportunity to work with other professionals) were found to be related to the level of work excitement among nurses. Results of this study suggest that enhancement of the learning environment can lead to higher levels of work excitement. Nurses in staff development can use the knowledge obtained in this study to assist nurses to find excitement in their work through new learning and work behaviors. The results suggest the importance of unit-based learning opportunities and flexible work groups that include other health professionals. PMID- 8408859 TI - Teaching infant CPR to mothers of cocaine-positive infants. AB - Cocaine abuse, a major problem in our society, has increased in women of childbearing age. Because cocaine-positive infants are at greater risk for apnea/sudden infant death syndrome (SIDS), their mothers need to be instructed in infant CPR. This study compared the use of a computerized interactive video learning system to the traditional infant CPR teaching method (lecture, video, and return demonstration) to determine the most effective method. This study indicated that mothers of cocaine-exposed infants can learn infant CPR in the immediate postpartum period and that the traditional method was found to be more effective. One unexpected positive outcome was the increase in these mothers' self-esteem. PMID- 8408860 TI - Comparison of two methods for teaching advanced arrhythmias to nurses. AB - An experimental study to compare the effectiveness of assisted self-directed learning (ASD) and traditional lecture methods (TLM) in acquiring competency for electrocardiogram interpretation was conducted within a conceptual framework of developmental and adult learning theory (Erickson, Tomlin & Swain, 1983; Knowles, 1980). After two teaching units were designed and an EKG posttest was constructed (r = .75) from actual patient records, registered nurse volunteers (N = 19) were randomly assigned to groups for simultaneous instruction. Because of the small sample size, statistical procedures were used to determine the effectiveness of randomization and to justify statistical procedures. EKG post-test scores for the experimental group (ASD, M = 81) and the control group (TLM, M = 71) were significantly different (one-tailed t = 1.79; p = .045); however, participants' prior knowledge of basic arrhythmias was not independent of method (ANCOVA, F[1, 16] = 21.373, p = .0003, covariate; F[1.16] = 2.312, p = .1479, main effects). EKG post-test scores were not related to critical care experience, cardiac monitoring experience, education level, or course satisfaction. While both methods are effective, ASD had higher mean scores and is more cost- and time efficient. PMID- 8408861 TI - Working effectively with LPN-RN orientees. AB - New LPN-RN graduates have hospital orientation needs that differ from those of generic RN graduates due to the differences between LPN and RN levels of practice. This article discusses potential problems related to the change from the LPN to the RN role and offers strategies to facilitate the transition. PMID- 8408862 TI - Enhancing presentation dynamics through voice projection. AB - Teaching adult learners requires ongoing preparation and commitment to excellence by adult educators. Central to the educational process is the educator's teaching style. One of the most powerful yet frequently underdeveloped qualities is the educator's voice. The authors present the Voice Projection Instrument as a personal development tool for adult educators to assess and enhance their speaking effectiveness in the classroom. PMID- 8408863 TI - Crossword: universal precautions. PMID- 8408864 TI - Study of mechanism of lipolysis inhibition by bovine milk proteose-peptone component 3. AB - Milk component 3 was an inhibitor of lipoprotein lipase activity responsible for spontaneous lipolysis occurring in milk stored at 4 degrees C. Experiments using a pH-stat apparatus and emulsified tributyrin showed that component 3 inhibited porcine pancreatic lipase. The lipolytic activity was fully restored by addition of sodium taurodeoxycholate and colipase to the emulsion containing component 3. Inhibition did not seem to be the result of a direct interaction between component 3 and the enzyme. Component 3 had a strong adsorption power superior to that of pancreatic lipase, as shown by tensiometric measurements at an n tetradecane-water interface. Lipase inhibition by component 3 could be the consequence of a rapid diffusion and preferential adsorption of component 3 at the oil-water interface provoking an important decrease of interfacial tension and avoiding the adsorption of lipase. PMID- 8408865 TI - Influence of milk proteins on the thermostability of the lipase from Pseudomonas fluorescens 33. AB - The effects of some milk proteins on the thermostability of the lipase from Pseudomonas fluorescens 33 were investigated. All purified milk protein fractions except kappa-casein that dissolved in phosphate buffer were effective for thermostabilization of the lipase. Thermal behavior of the lipase containing beta lactoglobulin was so specific that, after heating at 80 to 90 degrees C, activity remained high and was comparable with that of unheated treatment. The thermostability of the lipase containing whey proteins in synthetic salts solution was extensively lowered, but that containing casein micelles retained 50% of original activity after heat treatment at 80 degrees C for 10 min. Low temperature inactivation of the lipase was influenced by concomitant milk proteins. PMID- 8408866 TI - Antimicrobial susceptibility of bifidobacteria. AB - The antimicrobial susceptibility of 37 strains of bifidobacteria to 18 antimicrobial agents was determined by a macrodilution broth method. Most of the strains used were isolated from commercial yogurts and starters. Tested organisms were usually sensitive to Gram-positive spectrum antibiotics (bacitracin, erythromycin, lincomycin, and vancomycin), and most of the organisms were inhibited by a concentration < 1.56 micrograms/ml. Erythromycin was the most active agent; all strains were inhibited by < .19 microgram/ml. beta-Lactam antibiotics (penicillin G, ampicillin, methicillin, and cephalothin), showing a wide range of minimum inhibitory concentration, were less effective than Gram positive spectrum antibiotics. Most strains were somewhat resistant to cephalothin, exhibiting inhibition at concentrations of 6.25 to 25.0 micrograms/ml. Test organisms were most resistant to kanamycin, neomycin, paromomycin sulfate, nalidixic acid, and polymyxin B sulfate; inhibition occurred only at > or = 50 micrograms/ml, and strains were somewhat less resistant to gentamicin and streptomycin. Susceptibility to nitrofurantoin and tetracycline was variable; minimum inhibitory concentrations ranged from 1.56 to 50.0 and .39 to 50.0 micrograms/ml, respectively, but chloramphenicol had a narrow range from 1.56 to 6.25 micrograms/ml. PMID- 8408867 TI - Retinoid-induced modulation of immunoglobulin M secretion by bovine mononuclear leukocytes in vitro. AB - Effects of trans and cis isomers of retinol and retinoic acid on IgM secretion by bovine peripheral blood mononuclear leukocytes were evaluated in vitro. Mononuclear leukocyte cultures that were unstimulated or stimulated by pokeweed mitogen were supplemented with isomers of retinol and retinoic acid at 10(-10) to 10(-6) M. Concentrations of polyclonal IgM in supernatants from 14-d cultures were measured by an ELISA. Cultures stimulated by pokeweed mitogen consistently secreted more IgM than parallel, unstimulated cultures. Retinoid supplementation did not affect basal IgM secretion by unstimulated cultures. However, each retinoid affected IgM secretion by cultures stimulated by mitogen. The nature of the effect was dependent on the concentration of the specific retinoid. All-trans retinoic acid enhanced secretion at 10(-10) M and inhibited secretion at 10(-6) M. The other retinoids, however, did not inhibit IgM secretion at any concentration. Each retinoid enhanced IgM secretion at one or more concentrations, although enhancement was produced by much lower concentrations of retinoic acid isomers than retinol isomers. These results indicate that retinol and retinoic acid modulate polyclonal IgM secretion by cultures of bovine mononuclear leukocytes stimulated by mitogen. Future research will determine which subsets of the mononuclear leukocyte population are affected and whether trans-retinoic acid is the metabolite that produces these effects. PMID- 8408868 TI - Controlled ruminal infusion of sodium bicarbonate. 3. Influence of infusion dose on systemic acid-base status, minerals, and ruminal milieu. AB - Four ruminally cannulated, lactating Holstein cows were assigned to a 4 x 4 Latin square to monitor effects of intraruminal NaHCO3 infusion on temporal changes in ruminal and systemic acid-base status and mineral metabolism. Twice daily from 2 to 4 h postfeeding, cows were infused with 0, 110, 220, or 330 g of NaHCO3 dissolved in 3.8 L of water. All cows had access to their TMR of sorghum silage and concentrate (35: 65, DM basis) for 2 h twice daily. Ruminal fluid, blood, and urine were collected at feeding and every 30 min postfeeding for 12 h on the last day of each 14-d period. Total urine volume also was measured during this interval. Infusion of buffer increased ruminal fluid buffering capacity transiently at 4.5 h postfeeding but otherwise did not markedly affect ruminal fluid acid-base status. Systemic acid-base status was unaffected by the buffer primarily because renal excretion of base successfully reduced systemic base load. Urine volume increased in response to NaHCO3 infusion. Buffer infusion increased urinary excretion of Na, Mg, and K but decreased Ca excretion for 12 h postfeeding; Cl excretion was not affected. Buffer infusion tended to increase total VFA in ruminal fluid. Our data indicate that homeostatic mechanisms can eliminate exogenous base via the kidneys; hence, acid-base status was not perturbed by infusion of NaHCO3. The increased excretion of Mg and K with buffer infusion indicates that the dietary requirements for these minerals may be increased by NaHCO3. Although loss of Ca through the urine was reduced by buffer infusion, this reduction may indicate reduced availability of Ca to the cow. The diuresis accompanying large doses of NaHCO3 may increase dietary requirements for some minerals. PMID- 8408869 TI - Correlation of the alpha-lactalbumin (+15) polymorphism to milk production and milk composition of Holsteins. AB - The alpha-lactalbumin (+15) polymorphism (a single base variation 15 basepairs 3' of the alpha-lactalbumin transcription start point) was examined for its usefulness as a genetic marker for Holsteins. The +15 polymorphism is located in a region of the gene that is potentially involved in the regulation of alpha lactalbumin gene expression. Animals from two dairy herds and young sires from progeny-testing programs of four AI organizations were used in the analysis. A group of sons from a heterozygous sire were also evaluated. Each individual animal was genotyped at the alpha-lactalbumin (+15) locus, and differences of genotypes were investigated. Estimated differences among alleles were calculated for PTA for milk, kilograms of protein, protein percentage, protein dollars, kilograms of fat, fat percentage, and fat dollars. Animals having the alpha lactalbumin (+15) AA (an adenine on both alleles at position +15) genotype had statistically higher PTA for milk, kilograms of protein, protein dollars, kilograms of fat, and fat dollars than did the alpha-lactalbumin (+15) BB (a cytosine, guanine, or thymine on both alleles at position +15) animals. The alpha lactalbumin (+15) BB animals had higher protein and fat percentages than the alpha-lactalbumin (+15) AA animals. Animals that were heterozygous at this locus, alpha-lactalbumin (+15) AB, had intermediate values for all traits analyzed. These results indicate a potential marker or actual locus effect of the alpha lactalbumin (+15) polymorphism in Holstein cattle. PMID- 8408870 TI - Antibiosis revisited: bacteriocins produced by dairy starter cultures. AB - Well before the existence of starter bacteria was recognized, their activities were instrumental in preserving dairy foods. During growth in fermented products, dairy starters, including lactobacilli, lactococci, leuconostocs, streptococci, and propionibacteria, produce inhibitory metabolites. Inhibitors include broad spectrum antagonists, organic acids, diacetyl, and hydrogen peroxide. Some starters also produce bacteriocins or bactericidal proteins active against species that usually are related closely to the producer culture. Several bacteriocins have been biochemically and genetically characterized. Evaluating properties of the Lactobacillus acidophilus bacteriocin, lactacin B, led to a new purification protocol. Purified lactacin B migrates in SDS-PAGE as a single 8100 Da band with inhibitory activity after Coomassie blue staining. Production of lactacin B is enhanced by cultivation of the producer with the sensitive indicator, Lactobacillus delbrueckii ssp. lactis 4797; understanding this interaction may increase knowledge of production of bacteriocins in heterogeneous cultures. Bacteriocins have been recently identified in dairy propionibacteria. Jenseniin G, a bacteriocin produced by Propionibacterium jensenii P126, has narrow activity; propionicin PLG-1 produced by Propionibacterium thoenii P127 inhibits propionibacteria, some fungi, Campylobacter jejuni, and additional pathogens. Better understanding of these antagonists may lead to targeted biocontrol of spoilage flora and foodborne pathogens. PMID- 8408871 TI - The role of biological response modifiers in disease control. AB - Immune responses to infectious agents involve a complex set of interactions between cells and the factors they produce that culminate with disease resolution or death. Therefore, the manipulation of the immune system may have a great impact on the preservation and restoration of animal health. Biological response modifiers are agents that modify the host's response to pathogens with resultant beneficial prophylactic or therapeutic effects. The best known example of biological response modifiers are vaccines, by which administration of a nonpathogenic form of a microorganism prepares the immune system to produce a more effective response upon subsequent infection with the pathogenic form. Nevertheless, the use of biological response modifiers other than vaccines that enhance the immune response is now the focus of many investigations. In this review, a brief overview of the immune system is presented, and special emphasis is placed on possible areas of intervention with biological response modifiers and a description of the prophylactic and therapeutic potential of biological response modifiers. Specific examples are presented to demonstrate examples of disease modification by biological response modifiers through stimulation of nonspecific and antigen specific immunities. PMID- 8408872 TI - Recent advances in bovine vaccine technology. AB - A description of new commercial and experimental vaccines for viral and bacterial diseases of cattle can be broadly divided into those used for both beef and dairy cows and those used predominantly in dairy cattle. For both types of cattle, newer and experimental vaccines are directed against several of the important viral (e.g., bovine herpesvirus 1, bovine viral diarrhea virus, bovine respiratory syncytial virus, parainfluenza type 3, and foot-and-mouth disease virus) and bacterial pathogens (e.g., Pasteurella spp., Haemophilus somnus). The viral vaccines include gene-deleted, modified live, subunit, and peptide antigens. Newer bacterial vaccines, particularly those for Pasteurella spp., are composed of either modified-live vaccines or bacterins supplemented with toxoid or surface antigens. Haemophilus somnus vaccine research has concentrated mainly on defining unique surface antigens. Novel dairy cow vaccines would include the lipopolysaccharide-core (J5) antigen approach, which has been used for successful immunization against coliform mastitis. Core antigen vaccines also have reduced calf mortality from Gram-negative pathogens. Staphylococcal mastitis vaccines that contain capsular antigens, toxoids, or the staphylococcal fibronectin receptor are of active research interest. Vaccines against mastitis induced by Streptococcus agalactiae and Streptococcus uberis also are areas of intensive research. Delivery of multiple subunit antigens with optimal immune response induction has led to the investigation of attenuated heterologous viral and bacterial expression vectors such as bovine herpesvirus 1, vaccinia, and Salmonella spp. This discussion also demonstrates that molecular biology is being used to advance bovine vaccine technology. PMID- 8408873 TI - Somatolactogens, somatomedins, and immunity. AB - The neuroendocrine and immune systems participate as active partners in host homeostatic and defense mechanisms. This partnership involves a complex intercommunication system employing an array of shared ligands and receptors. Hormones of the somatolactogen family have marked influences on immune events in vivo, including the maintenance of lymphoid tissue cellularity, the promotion of DNA synthesis in these tissues, and the stimulation of a number of immune effector mechanisms. Both growth hormone and prolactin function to promote erythropoiesis and DNA synthesis in bone marrow precursors. Our results have shown that the somatolactogens and a member of the somatomedin family, IGF-I, are particularly effective in modulating the effector functions in phagocytic cells, including the production of reactive oxygen intermediates and tumor necrosis factor-alpha and the oxygen-dependent killing of bacteria. Evidence indicating a role of IGF-I in modulating immune functions is more recent but nonetheless compelling. Accumulated data suggest that somatolactogenic hormones, as well as one member of the somatomedins, are produced by cells of the immune system and can regulate local immune events. Although the molecular mechanisms by which the somatolactogens and somatomedins exert their effects on immune tissues are only now being explored, the pleiotropic nature of these effects suggests that these hormones participate at endocrine, paracrine, and perhaps autocrine sites of action. PMID- 8408874 TI - An urgent plea for a standardized bonding (adhesion) test. PMID- 8408875 TI - Regaining of OH group in dehydroxylated hydroxyapatite. PMID- 8408876 TI - Communicating new information in dentistry. PMID- 8408877 TI - Esthetic dentistry--a health service. PMID- 8408878 TI - Membrane protein expression by Actinobacillus actinomycetemcomitans in response to iron availability. AB - Iron-limited growth conditions under anaerobiosis were established for Actinobacillus actinomycetemcomitans strains Y4, JP2, and 75 by use of the ferrous ion chelator 2,2'-dipyridyl. Growth inhibition was reversible with both ferrous and ferric iron sources. Sarcosyl-insoluble membrane fractions of iron stressed anaerobic A. actinomycetemcomitans cultures revealed a similar iron repressible protein of approximately 70 kDa in A. actinomycetemcomitans strains Y4, JP2, and 75. This 70-kDa protein was recognized by serum from localized juvenile periodontitis patients and a periodontally healthy subject. This suggested that the 70-kDa iron-repressible protein may be expressed in vivo. When A. actinomycetemcomitans was grown under aerobic conditions, the ferric iron chelator, ethylenediamine di(o-hydroxyphenylacetic acid) (EDDA) was utilized for growth limitation. EDDA inhibition was reversible in strain Y4 with ferrous and ferric iron sources. An iron-repressible protein of approximately 70 kDa was also noted in iron-stressed aerobic cultures. The 70-kDa protein may be involved in iron transport by A. actinomycetemcomitans. Preliminary experiments were performed to examine potential iron transport systems for A. actinomycetemcomitans. Production of the two most common chemical types of siderophore was not detected in A. actinomycetemcomitans culture supernatants. Iron-starved A. actinomycetemcomitans cells did not bind transferrin or lactoferrin in a dot blot assay. PMID- 8408879 TI - Antigenic relationships among oral Actinomyces isolates, Actinomyces naeslundii genospecies 1 and 2, Actinomyces howellii, Actinomyces denticolens, and Actinomyces slackii. AB - Antigenic relatedness among human strains of oral Actinomyces and similar isolates from cattle has been analyzed by agglutination and immunoblotting. Whole cell agglutination placed A. viscosus serotype II, A. naeslundii serotypes II and III, Actinomyces NV, and strains from numerical taxomonic clusters C1, C2, C3, C4, and C6 into a single group. A. viscosus serotype I cross-reacted weakly with this group. A naeslundii serotype I strains and the cattle isolates Actinomyces denticolens and Actinomyces howellii were distinct. The agglutination results for A. slackii were equivocal. Immunoblots of cell wall extracts developed with non absorbed sera showed cross-reactivity (23% to 90% antigenic similarity) among all of the strains tested, including A. israelii. The range of antigenic similarities among the group which included strains of A. viscosus serotype II, the A. naeslundii serotypes, and clusters C1, C2, C3, C4, and C6 was from 39% to 89%. Immunoblotting showed that A. howellii and A. denticolens were between 39% and 72% similar to A. naeslundii and A. viscosus. Absorption of antisera with A. israelii cell walls removed antibodies recognizing antigens common to Actinomyces and made the sera more specific. Immunoblotting with absorbed sera supported the grouping and separation of strains shown by agglutination. In some cases, serotypes could be included into a specific taxonomic cluster. A. naeslundii serotype II and Actinomyces NV most closely resembled cluster C1 strains, and A. naeslundii serotype III resembled cluster C1 strains, and A. naeslundii serotype I and A. viscosus serotype I were included into clusters C5 and C7, respectively. The results support a recent proposal that strains of A. viscosus serotype II, A. naeslundii serotypes II and III, and Actinomyces NV be included into A. naeslundii genospecies 2, that A. naeslundii serotype I should be designated A. naeslundii genospecies 1, and that A. viscosus serotype I should be retained distinct from A. naeslundii, as A. viscosus. PMID- 8408880 TI - Transport of sugars, including sucrose, by the msm transport system of Streptococcus mutans. AB - The range of substrates transported by the sugar-binding protein-dependent msm (multiple sugar metabolism) system of S. mutans was investigated. By determining the ability of unlabeled sugar to compete with radiolabeled melibiose transport, we have demonstrated that the transported sugars included a number of carbohydrates structurally related to raffinose. A model accommodating these results has been devised which accounts for the sugars transported by the msm transport system. Competition with radiolabeled melibiose transport indicated sucrose to be an msm substrate. This was confirmed by examination of uptake of radiolabeled sucrose in scrAB mutants lacking the sucrose-specific phosphotransferase system. PMID- 8408881 TI - An in vitro stimulation of the effects of chewing sugar-free and sugar-containing chewing gums on pH changes in dental plaque. AB - The objective of these studies was to simulate the effect of chewing sugar-free and sucrose-containing chewing gums on the return of the pH to neutrality after exposure to sucrose of plaque located on the buccal (BLM) and lingual (LLM) surfaces of the lower molar teeth. In study 1, a 0.5-mm-deep artificial plaque containing Streptococcus oralis cells was exposed to 10% sucrose for one min, and a 0.1-mm-thick film of sucrose-free artificial saliva was then flowed over the plaque surface at the unstimulated salivary film velocities previously found at the BLM and LLM sites. At the time of the pH minimum (pH 4-5), one of three conditions was simulated: (a) a no-gum-chewing control, or chewing for 20 min on either (b) a sugar-free gum or (c) a sucrose-containing gum. The recovery of the plaque pH to resting values was rapid during simulation of chewing a sugar-free gum (SFG), much slower with the no-gum control, and even slower with simulation of chewing a sucrose-containing gum (SCG). The pH recovery was slower with the BLM than the LLM plaque. In study 2, the BLM plaque was exposed to a 2% sucrose solution for 20 min under stimulated salivary conditions, to simulate the consumption of a meal, followed by one of conditions (a), (b), or (c) described above. The pH recovery with simulation of chewing a SCG was faster than with the no-gum control, but much slower than with the SFG simulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408882 TI - Comparison of tooth surface-specific dental caries attack patterns in US schoolchildren from two national surveys. AB - The 1979-1980 and the 1986-1987 National Institute of Dental Research (NIDR) surveys of school-aged children revealed that virtually all tooth surfaces experienced a decrease in caries prevalence during the inter-survey period. Overall, there was a 28% decrease in the proportion of tooth surfaces attacked by caries for the primary dentition between the two surveys. The decrease for primary incisors was numerically small (5 surfaces per thousand surfaces at risk) and not statistically significant, whereas decreases in the canines and primary molars were considerably larger (23 surfaces per thousand) and statistically significant. For the permanent dentition, the overall decrease in the proportion of surfaces attacked was 35% during the 1979-87 period. Differences between the two surveys in the proportions of surfaces with caries were largest for pit and fissure surfaces (56 surfaces per thousand), followed by those for posterior approximal surfaces (14 surfaces per thousand) and all other smooth surfaces (5 surfaces per thousand). Almost all of these differences were statistically significant, except for some surfaces which experienced very few caries. PMID- 8408883 TI - Effects of CO2 laser irradiation in vivo on rat alveolar bone and incisor enamel, dentin, and pulp. AB - Previous studies have shown that a surgical 'window' can be drilled in alveolar bone for experimental manipulation of the underlying enamel organ and enamel. To determine whether similar and/or improved access could be obtained by use of the surgical capabilities of laser optics, and to note the effects of laser irradiation in vivo on the extracellular matrices and cells of bone, enamel, and dentin, tissue responses to laser-created lesions were examined histologically. Briefly, samples were prepared in which the alveolar bone along the inferior mandibular border of Wistar rats was exposed, and a continuous-wave CO2 laser equipped with a custom-made micromanipulator was used to penetrate the bone and to create lesions within the lower incisor. Animals were perfusion-fixed at either 10 min or 10 days post-treatment, and affected tissues were processed for light and transmission electron microscopy. At 10 min, all lesions consisted of a void of ablated tissue containing some organic debris. Tissues immediately surrounding the lesion were generally intact, but showed some damage, presumably resulting from elevated temperature effects. At 10 days, lesions in the bone, dentin, odontoblast layer, or pulp showed morphological evidence of tissue repair represented by the presence of cell infiltrates, new bone, or reparative dentin. In lesions that were created during the secretory stage of amelogenesis that had moved into the maturation stage, there was evidence of delayed or incomplete maturation of enamel (i.e., retention of organic matrix normally lost during maturation) related to the enamel organ affected by the laser treatment. In the bone lesion at 10 days, new bone formation was observed, while bone fragments originally created at the time of lasing were surrounded by mononuclear and large multinucleated giant cells. It is thus concluded that the application of this laser system is an alternative method for exposing unerupted dental tissues for experimental manipulation, and that laser irradiation may also be useful for the study of mineralized tissue repair. PMID- 8408884 TI - Adsorption of low-molecular-weight sodium polyacrylate on hydroxyapatite. AB - Adsorption of low-molecular-weight sodium polyacrylate from aqueous solution onto synthetic hydroxyapatite was studied at room temperature so that the mechanism of adhesion of polyacrylate cements to tooth mineral could be elucidated. The adsorption isotherm of sodium polyacrylate was Langmuirian in shape and was thus qualitatively different from that of polyacrylic acid (Misra, 1991), which exhibited an adsorption maximum. The self-association of the molecules that probably causes the maximum to occur with polyacrylic acid was effectively absent for the relatively well-ionized, electrostatically repelling polyacrylate ions of the salt. With the adsorption of acrylate ions, the concentration of phosphate ions increased monotonically, while the concentration of calcium ions showed a minimum. The adsorption of sodium polyacrylate was irreversible, as it was for polyacrylic acid. PMID- 8408885 TI - Chemical characterization of the resin-dentin interface by micro-Raman spectroscopy. AB - The chemical nature of the interface between dentin and adhesive resin materials was characterized by micro-Raman spectroscopy. The resulting chemical profiles were correlated with photomicrographs obtained by SEM after an argon-ion-beam etching treatment of the sample surface. Two commercially available dentin adhesive systems, of which one was also applied with a different conditioning agent, were investigated. Raman spectra, which were recorded along line scans across the interface with a step increment of 1 micron, revealed that resin effectively penetrated 4 to 6 microns deep into the superficially decalcified dentin zone. Across the interface, a gradual transition from resin to dentin over the interdiffusion zone with a mixed contribution of both substances was noticed. Finally, resin appeared to penetrate to the entire decalcification depth of dentin regardless of the aggressiveness of the conditioning procedure. PMID- 8408886 TI - Enhanced chemical strengthening of feldspathic dental porcelain. AB - Previous studies on ion exchange of dental ceramics have shown that the biaxial flexural strength can be improved by exchanging potassium for sodium ions at temperatures below the strain point. The rubidium ion is bigger than the potassium ion and can also be considered as a candidate for replacing smaller ions, i.e., sodium or potassium, although it has not been used for dental ceramics. The double-step method uses the exchange of a small ion for a large ion (Li for Na) above the strain point, and the further exchange of a large ion for a small ion (K for Li) below the strain point. The purpose of this study was to compare the effect of rubidium-for-potassium ion exchange with that of potassium for-sodium exchange (Tuf-Coat, G-C International Corp., Japan) on the flexural strength of a feldspathic dental porcelain and to test the hypothesis that a double-step ion exchange can lead to greater strengthening than potassium ion exchange alone. Weight measurements were performed before and after treatment. Qualitative chemical analyses allowed the rubidium, potassium, and sodium concentration profiles to be determined along cross-sections of the specimens. The maximum biaxial stresses were calculated after specimens were fractured in water on a ball-on-ring fixture at 0.5 mm/min. Relative to the untreated control group, the flexural strength of the potassium-exchanged groups was significantly increased, except for those treated at 400 or 500 degrees C. All the groups treated with RbNO3 exhibited a significant increase in flexural strength, with a maximum for the group treated at 450 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408887 TI - Assessment by nano-indentation of the hardness and elasticity of the resin-dentin bonding area. AB - The hardness and Young's modulus of the successive layers across a resin-dentin bonding area were determined by nano-indentation for four commercially-available dentin adhesive systems, of which two were also applied with a different conditioning agent. With a computer-controlled nano-indentation technique, minute triangular indentations were made within a small area of a few micrometers' diameter at a load of a few milli-Newtons. The load and displacement of the indenter were continuously monitored during the loading-unloading sequence, so hardness and Young's modulus could be computed as a function of the indenter geometry and the applied load. The hardness of the resin-dentin interdiffusion zone was significantly lower than that of unaltered dentin. A gradient of moduli of elasticity was observed from the rather stiff dentin over a more elastic resin dentin interdiffusion zone and adhesive resin layer to the restorative composite. That gradient was more substantial in those systems that produced relatively thick adhesive resin layers or supplementally provided a filled low-viscosity resin as an intermediate layer between the adhesive resin and the bulk restorative composite. Such an elastic bonding area might have a strain capacity sufficient to relieve stresses between the shrinking composite restoration and the rigid dentin substrate, thereby improving the conservation of the dentin bond and, as a consequence, the marginal integrity and retention of the restoration. PMID- 8408888 TI - Diagnostic value of orthopedic tests in patients with temporomandibular disorders. AB - For a better definition of diagnostic subgroups of patients with temporomandibular disorders (TMD), clinical orthopedic tests have been developed for the masticatory system, for use together with the commonly used active movement tests and palpation. In the present study, the characteristics and additional diagnostic value of four orthopedic tests--namely, passive opening, the joint play test, compression, and the static pain test--were determined for a patient group, diagnostic subgroups, and a control group. Significant differences among the scores of the tests in the different groups indicated that all six orthopedic tests contributed to the diagnostic process. Because of the close functional relationship of the masticatory system, most information was obtained when patients indicated where pain occurred during the different tests. Logistic regression techniques were used for determination of the relative diagnostic value of each orthopedic test in the diagnostic process. Active movement was the most powerful test for distinguishing the different subgroups. Palpation and passive opening were additionally useful for distinguishing between patients and control subjects and between the subgroups of arthrogenous and myogenous patients. Besides active movement, both compression and the joint play test played a minor role in the distinction of subgroups of arthrogenous patients. It may be concluded that in routine clinical practice, besides history taking and conventional radiography, a functional examination consisting of active movements, passive opening, and palpation provides valuable diagnostic information. In patients with a specific diagnostic problem, one of the other additional tests might be indicated. PMID- 8408889 TI - Women's health: a national plan for action. AB - Women's health is a central item on the national health agenda for the 1990s. Renewed attention to the health needs of women has led to a national plan for action with interventions in research, prevention, treatment, training, public information, and public policy. Since research and training are pivotal to the promulgation of any national health agenda, this has particular importance for dental research and dental education. The 38-goal PHS Action Plan for Women's Health, which directs resources of the U.S. Public Health Service (PHS) toward women's health is coordinated by the Office on Women's Health and implemented through the PHS agencies and offices including the National Institutes of Health (NIH), Centers for Disease Control and Prevention (CDC), and Food and Drug Administration (FDA). It encompasses large scale research interventions like the NIH "Women's Health Initiative" as well as other biomedical, biobehavioral, and health services research programs. PMID- 8408890 TI - Beyond pregnancy gingivitis: bringing a new focus to women's oral health. AB - The compromised functional status, physical confinement, medical conditions, and cognitive impairments of significant numbers of women have important implications for oral health risk and dental treatments. In addition, there are a variety of economic, social, psychological, and behavioral factors which are operative in placing women at high risk for development of oral diseases. This paper describes the available U.S. data on women's oral health and examines some of the biological, behavioral, and societal factors which may be important for a more comprehensive understanding of this subject. It is this broad array of factors which differentiate women from men and which should be considered when defining priority research and treatment issues for women's oral health. PMID- 8408891 TI - Health services and women's oral health. AB - Apart from tooth loss and edentulism, data on the relative levels of oral health of men and women are scarce. Evidence from Europe and to a lesser extent from the U.S. shows that women have fewer natural teeth present than men and have higher levels of edentulism. In Europe there is some evidence that socially deprived rural women have the highest levels of edentulism. Due to the lack of adequate detail in published reports of studies, it is difficult to relate the oral health of women with the availability, acceptability, and accessibility of dental services. Women tend to attend for dental care more regularly than men though there is some evidence that women are more fearful of dental treatment and also perceive cost as a barrier to dental care. Data from the Republic of Ireland suggest that women working in the home have higher levels of tooth loss and edentulism than those working outside the home. There is a need for consensus on methods of reporting oral health data in major descriptive studies. Further research is required to investigate why women tend to lose their natural teeth at an earlier age than men. PMID- 8408892 TI - Gender differences in oral health research: beyond the dichotomous variable. PMID- 8408893 TI - Women's health: implications for health professional education. PMID- 8408894 TI - Non-surgical gingival curettage in dental hygiene curricula. PMID- 8408895 TI - Comparison of two instructional approaches in preclinical dental hygiene. PMID- 8408896 TI - Licensure--a house divided. PMID- 8408897 TI - Curriculum guidelines for practice administration. Section on Practice Administration of the American Association of Dental Schools. PMID- 8408898 TI - A preliminary study of the effect of eliminating requirements on clinical performance. AB - This study determined the effect of a clinical program driven by patient needs upon students' productivity, attitudes, and academic performance. A group of eight senior students, whose academic and clinical performance profile replicated that of the rest of the class, were chosen to participate in a year-long non requirement clinic. The students were expected to attend all clinic sessions, and treat their assigned patients. Their performance was compared to that of classmates in the regular requirement-driven curriculum. The non-requirement group had significantly higher academic achievement and significantly outproduced their classmates. Non-requirement students had no state board failures, versus 17 percent in the regular curriculum, and reported significantly lower stress. This study suggests that predoctoral clinical programs can maintain quality and productivity in the absence of unit requirements. PMID- 8408899 TI - Predictors of dental students' belief in the right to refuse treatment to HIV positive patients. AB - We examined predictors of dental student's belief that they should be allowed to refuse treatment to HIV-infected persons. We surveyed 181 first; second-, and fourth-year dental students at a large urban university using a 44-item, self administered anonymous questionnaire and a measure of dispositional optimism. Several composite measures were created and their relationship to belief in the right to refuse treatment was assessed. Regression techniques were used to describe the relationship between the dependent and independent variables. Results indicated that non-professional attitudes, low optimism scores, low levels of comfort with homosexuality, and gender were the best predictors of belief in the right to refuse treatment to HIV-infected patients. Neither knowledge of HIV, year in dental school, or fear of contagion reliably predicted belief in the right to refuse treatment. PMID- 8408900 TI - Student satisfaction with attending systems. PMID- 8408901 TI - The association of gender with clinic activity and broken appointments. PMID- 8408902 TI - A problem-based course in dental implantology. AB - A problem-based predoctoral implantology course was implemented for ten senior dental students at the University of Texas Health Science Center at San Antonio. Students met in small tutorial groups three times a week to analyze patient cases developed for the course. As students explored the patient's problems, they identified areas where their knowledge was deficient, as well as special interests, and prioritized these areas, known as learning issues, into a learning agenda for subsequent study. During unprogrammed free time between tutorials, students researched the learning issues, and came to the next tutorial prepared to discuss what they had learned and in so doing, helped their peers learn the material. The faculty tutor used non-directive questioning to facilitate group problem-solving and promote interaction but did not lecture or give answers. The cases were developed in the "progressive disclosure paperback" format, reflecting the evolving nature of the patient's implant therapy. In conjunction with the PBL tutorials, students performed implant surgery and prosthodontics on a minimum of three patients and provided maintenance therapy on three to four other implant patients. Course evaluation consisted of three elements: (1) weekly peer evaluation by the students using a six item checklist, (2) tutor evaluation, using the same checklist, and (3) an oral examination. The tutor met individually with each student midway through the course and at the conclusion to summarize the peer evaluations and discuss the student's performance, including contributions to group process. Student reaction to this PBL experience was enthusiastically supportive, and student performance exceeded faculty expectations. PMID- 8408904 TI - Chronology of U.S. Dental School Establishment. PMID- 8408903 TI - Meeting challenges in dental hygiene through cooperative education. PMID- 8408905 TI - Curriculum guidelines for orthodontics. AADS Section on Orthodontics and the Council on Orthodontic Education of the American Association of Orthodontists. PMID- 8408906 TI - Curriculum guidelines for periodontics for dental hygiene programs. The curriculum guidelines committee of the AADS Section on Dental Hygiene Education. PMID- 8408907 TI - Plugging the vascular leak. PMID- 8408908 TI - Treatment of facial telangiectasia with sclerotherapy, laser surgery, and/or electrodesiccation: a review. AB - BACKGROUND: Facial telangiectasia is extremely common in patients with fair complexion. Electrosurgery has been the primary form of treatment in the past. OBJECTIVES: The authors retrospectively evaluated over 300 patients treated with sclerotherapy for facial telangiectasias. METHODS: Retrospective analysis of over 300 patients with facial telangiectasia treated with sclerotherapy and 55 facial spider telangiectasias treated with laser. RESULTS: Sclerotherapy utilizing hypertonic saline, Sclerodex, polidocanol, and sodium tetradecyl sulfate resulted in-between 50 and 100% resolution of facial telangiectasia with minimal complications. CONCLUSION: The treatment of facial telangiectasia should include sclerotherapy and laser treatment modalities, which effect a higher rate of clearance with less adverse sequela. PMID- 8408909 TI - Topical silicone gel sheeting in the treatment of hypertrophic scars and keloids. A dermatologic experience. AB - BACKGROUND: Topical silicone gel sheeting has been used successfully in the management of hypertrophic and keloid scars resulting from thermal burn wounds. METHODS: An open-labelled approach using the silicone gel sheets was performed using hypertrophic and keloid scars secondary to surgical procedures or traumatic insults. RESULTS: The silicone gel sheets resulted in moderate improvement in scar thickness, scar color and was noted to be effective to some degree in all tested. The material was easy to use and painless. CONCLUSION: Topical silicone gel sheeting is an effective method for the treatment of hypertrophic and keloid scars and may be considered useful in the treatment of these difficult cutaneous lesions. PMID- 8408910 TI - Recurrent basal cell carcinoma of the back infiltrating the spine. Recurrent basal cell carcinoma. AB - BACKGROUND: Risk of basal cell carcinoma (BCC) recurrence appears to be related to tumor location, tumor size, treatment modality, radicalness of excision, and histologic type. Recurrences of BCC on the trunk and extremities are rare, with 97% of all recurrent lesions being located in the head and neck region. OBJECTIVE: This report concerns a man who had a BCC of the back removed with electrodesiccation and curettage in 1980 at the age of 36. In the following 10 years the patient experienced five recurrences and finally developed infiltration of the thoracic spine that precluded further attempts at radical excision of the neoplasm. METHODS AND RESULTS: The pathology of the BCC in our patient revealed a number of histologic features that have been associated with aggressive behaviour. These include an infiltrative growth pattern characterized by an irregular and acute tapered profile of the tumor cell groups and fibroblast-rich stroma, infiltrating invading tumour edges, poorly peripheral palisading, nuclear pleomorphism, and perineural invasion. CONCLUSION: Basal cell carcinomas that occur in sunlight-protected areas of the trunk of middle-aged individuals can occasionally exhibit aggressive and highly invasive features as well as a marked tendency for local uncontrollable recurrences. The prognosis for patients with this variety of BCC is poor, especially when progressive infiltration of deep tissues does not permit, as in our case, any possible further radical surgical procedure. PMID- 8408911 TI - Review of continuous sutures in dermatologic surgery. AB - BACKGROUND: Continuous sutures have been developed to close surgical defects to save time, to distribute tension evenly, or to improve the cosmetic result. The choice of suture technique can significantly affect the speed of closure and the eventual appearance of the scar. OBJECTIVE: To review the different types of continuous sutures, their advantages, and disadvantages. CONCLUSION: Recommendations are made regarding the appropriate use of continuous sutures in wounds depending on the amount of tension in the wound, and requirement of a good cosmetic result. PMID- 8408912 TI - Rational sclerotherapy techniques for leg telangiectasia. PMID- 8408913 TI - Anatomical examination of leg telangiectases with duplex scanning. AB - BACKGROUND: Earlier studies using Doppler ultrasound suggested that thigh telangiectasia might be associated with reflux in the larger subcutaneous veins. OBJECTIVE: The aim of the study was an accurate description of the anatomy of the venous drainage from thigh telangiectases and evaluation of the extent of venous valvular incompetence using a noninvasive imaging method. METHOD: High-definition color duplex ultrasonography was used to achieve the objective. RESULTS: Thirty seven legs with lateral and/or medial thigh telangiectasia were examined. Patients studied were those without saphenofemoral, saphenopopliteal, or deep venous abnormalities on Doppler ultrasound examination. Altogether 53 sites were tested. In 89% reticular vein incompetence was found close to telangiectases. Often reticular vein incompetence was associated with reflux in larger epifascial veins. In 15% incompetent perforating veins were detected between reticular veins and the deep venous system. CONCLUSION: An accurate, functional reconstruction of the subcutaneous venous drainage was accomplished. It was found that telangiectasia was rarely an isolated condition, but was usually associated with incompetence in other elements in the venous drainage of the subcutaneous tissue. PMID- 8408914 TI - Doppler ultrasound findings in reticular veins of the thigh subdermic lateral venous system and implications for sclerotherapy. AB - BACKGROUND: Treatment of varicose veins is directed towards eliminating sources of reflux causing venous hypertension in the superficial venous system. Doppler ultrasound is routinely used to localize sources of reflux prior to treatment of large varicose veins. OBJECTIVE: To assess the value of Doppler ultrasound in localizing reflux in subcutaneous reticular veins associated with groups of lateral thigh telangiectases. METHODS: Seven hundred patient records in which Doppler ultrasound was performed on thigh reticular veins were reviewed. Patients with reflux of the saphenofemoral junction, saphenopopliteal junction, or major saphenous system perforators were excluded. RESULTS: Audible reflux upon distal calf compression release was noted in thigh subcutaneous reticular veins of 618 out of 700 patients (88%). These incompetent reticular veins were associated with groups or webs of telangiectases and/or venulectases on the lateral thigh in almost all cases. Reflux was loudest just distal to the lateral femoral condyle. In 20 patients Doppler ultrasound was performed on a reticular vein while a telangiectasia associated with the reticular vein was injected with sclerosing solution. In 15 patients a clearly audible flow signal was heard in the reticular vein during injection. CONCLUSION: These findings indicate an association of telangiectatic veins with reticular veins on the thighs. Furthermore, this study underscores the value of Doppler in determining sources of reflux in smaller veins and implies that sclerotherapy of reticular veins is necessary for treatment of thigh telangiectatic webs. PMID- 8408915 TI - Inadvertent intra-arterial injection complicating ordinary and ultrasound-guided sclerotherapy. AB - BACKGROUND: Intra-arterial injection is the most dreaded complication of sclerotherapy of varicose veins. The medical literature does not contain enough data to enable one to formulate a definitive strategy for either the prevention or treatment of this complication. OBJECTIVE: Seven cases of intra-arterial injection are presented. The causes of these accidents, the recognition of the signs that intra-arterial injection has occurred, and the treatment of these cases are discussed. METHODS: Intra-arterial injection occurs in a variety of different settings. The signs that an intra-arterial injection has taken place are variable, but in some cases there are no signs until irreversible tissue damage has already set in. Treatment protocols are based mainly on anticoagulation with heparin, which is administered both intravenously and subcutaneously. RESULTS: Heparin, whether administered continuously by the intravenous route or subcutaneously (twice daily), makes affected areas look and feel better, but there is no proof that it alters the final outcome with respect to tissue necrosis. In agreement with previous authors, we believe that there is a beneficial effect. Coumadin, on the other hand, appears to be completely ineffective. CONCLUSION: Protocols employing heparin should be considered in cases of suspected intra-arterial injection. The length of time heparin should be continued is uncertain. The best "treatment" is prevention. PMID- 8408916 TI - Indications for the sclerosing agent polidocanol (aetoxisclerol dexo, aethoxisklerol kreussler) AB - BACKGROUND: Polidocanol is a widely used sclerosing agent that was first developed as a local anesthetic. In France it is used in varicose veins as well as in telangiectases. OBJECTIVE: To improve the knowledge of the effects of polidocanol and provide recommendations for better use. METHODS: Based on 12 years of experience in treating nearly 11,000 patients with polidocanol, the author reviews his observations. A theoretical model of dilution of sclerosing agents in injected veins is provided. RESULTS: The author observed: seven cases of minor urticaria, occurrence of grey veil when used under the air-block technique, one case of cutaneous necrosis (5 cm3), and some small epidermic necrosis (1-mm diameter) and neither shock nor severe adverse reaction. CONCLUSION: Polidocanol is characterized by its spasmogenic action on veins, excellent local tolerance, and a very low incidence of general reactions. Low concentrations (0.3 to 0.6%) can be used for the treatment of quite large varicose veins if an adequate volume is injected. PMID- 8408917 TI - Two-stage correction of depressed glabella and nasal root scar contracture utilizing subcutaneous tissue advancement flaps and a layered soft tissue/procerus muscle transposition flap. AB - BACKGROUND: A postoperative nasal root and glabellar forehead deformity comprised both vertical and horizontal elements, with scar contracture and fixation across the left medial nose base, extending into the adjacent superior canthal region. METHODS: A two-stage reconstruction was performed, initially correcting the vertical centrally placed forehead depression by dual subcutaneous flaps advanced centrally and joined. The horizontal nasal root contracture was repaired at a second stage utilizing a procerus muscle transposition flap elevated on the ipsilateral deep contracture side, imbricated in serpentine fashion on itself and sutured into the depression over the scar base. RESULT: Significant improvement followed. CONCLUSION: The careful and precise placement of subcutaneous tissue and muscle can serve to assist in filling certain deep defects and in elevating scar lines. PMID- 8408918 TI - Clinical manifestations of 174 AIDS cases in Maharaj Nakorn Chiang Mai Hospital. PMID- 8408919 TI - Clinical significance of serum adenosine deaminase activity in patients with mycosis fungoides. AB - Adenosine deaminase is one of the key enzymes in purine nucleotide degradation. This enzyme exists in most of the human tissues and the activity is high in lymphatic tissues, especially in T lymphocytes. Elevated adenosine deaminase activity in T cell leukemia has been reported, and its inhibitor, deoxycoformycin, has been developed as an antitumor agent. In some types of leukemia, serum adenosine deaminase activity increases in accordance with the severity of the disease. Although mycosis fungoides rarely involves peripheral blood, tumor cells do invade the skin. In order to evaluate the clinical significance of adenosine deaminase in mycosis fungoides, adenosine deaminase activity was measured in sera of 15 patients with mycosis fungoides at various stages. The mean enzyme activity was 23.2 IU/l, which was high with statistical significance compared with healthy controls (P < 0.001). Nine of twelve patients in the plaque stage (T2N0M0, IB) showed higher adenosine deaminase activity than did the normal population. The mean adenosine deaminase activity in sera in the patients in the plaque stage (T2N0M0, IB) was as high as 19.0 IU/l (range 13.7 21.4) with statistical significance compared with healthy control (P < 0.001). Three tumor stage patients without visceral involvement (T3N0M0, IIB) showed higher levels of adenosine deaminase activity (19.7, 21.5, 24.4 IU/l). An erythrodermic patient (T4N0M0, III) also had a high adenosine deaminase activity 28.4 IU/l. Two tumor stage patients with organ involvement (T3N0M1, IVB) exhibited extremely high adenosine deaminase activity (60.9, 32.2 IU/l). The adenosine deaminase activity in sera showed a tendency to become higher with the extension of the stages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408920 TI - Analysis of 70 KD heat shock protein (HSP70) expression in the lesional skin of lupus erythematosus (LE) and LE related diseases. AB - Biopsied specimens from skin lesions of SLE were studied for expression of 70 KD heat shock protein (HSP70). The pattern of HSP70 expression in SLE was diffuse in whole epidermis, hair follicles, and sweat gland cells and rather more intense than that in other control groups or normal skin. No significant differences in HSP70 expression were observed between sun-exposed and protected areas of SLE skin lesions. Unlike SLE, reduced or no expression of HSP70 was observed in skin lesions of DLE. In tissue culture, UVB radiation in vitro induced relatively intense expression of HSP70 in the nuclear area of keratinocytes. A few gamma delta T cell receptor positive cells which might respond to HSP70 expressing cells were detected in the basal layer of skin lesions of diseases. These studies suggest that aberrant expression of HSP70 in skin lesions of SLE might contribute to both skin lesions and antibody formation in SLE. PMID- 8408921 TI - Localized bullous pemphigoid: report of a case with an immunofluorescence and electron microscopical studies on the lesional distribution of 180-KD bullous pemphigoid antigen, beta 4 integrin, and type VII collagen. AB - A 67-year-old woman with a left-sided hemiplegia had localized bullous pemphigoid demonstrating typical clinical lesions on the left pretibial skin and the radial side skin of the right forearm. The histology showed a subepidermal blister with extensive hyperkeratosis, hypergranulosis, and acanthosis. Direct immunofluorescence revealed distinct linear deposits of IgG and C3 at the dermo epidermal junction in the perilesional skin and in the roof of the blisters, but few deposits in nonlesional skin. Electron microscopy revealed separation in the lamina lucida. Indirect immunofluorescence of type VII collagen showed its localization in the blister floor. The distribution of the 180-KD bullous pemphigoid antigen (BPA) and beta 4 integrin, hemidesmosomal transmembrane proteins, were studied in the lesional skin by indirect immunofluorescence. Both 180-KD BPA and beta 4 integrin were localized in the blister roof. By immunoelectron microscopy, beta 4 integrin was detected in small groups on the cell surface facing the blister cavity. Since the epitope of the monoclonal antibody to 180-KD BPA used here is known to be localized at a distance of 20 to 50 nm from the membrane surface and this epitope retained in the blister roof, it appears that the blister was produced in the deep lamina lucida. The lesions were cleared with topical 0.05% clobetasole propionate ointment. PMID- 8408922 TI - Langerhans' cells of elderly patients with decubital ulcers. AB - An ultrastructural and morphometric study compared Langerhans' cells in sacral epidermis, 8-10 cm from the lesion of patients (mean age 70) with decubital ulcers, with those in the patients' own normal epidermis from the upper leg and with those in the epidermis from the upper leg of normal age-matched volunteers. The Langerhans' cell section area was significantly lower in patients' control leg epidermis (25.86 +/- 2.29 microns2) than in that from normal controls (35.74 +/- 3.76 microns2) or that from patients' sacral epidermis near the lesion (41.26 +/- 3.45). The number of Langerhans' cell granules was higher in control leg epidermis of patients (10.22 +/- 1.26) and was significantly higher in lesioned sacral epidermis (12.94 +/- 1.90) than in normal controls (6.97 +/- 1.47). In Langerhans' cells in patients' epidermis, formative stages of Langerhans' cell granules were observed. The findings may indicate that Langerhans' cells in patients' epidermis are in an active state. PMID- 8408923 TI - Vitiligo, rheumatoid arthritis and pernicious anemia. AB - A patient with a 46-year history of vitiligo who also presented rheumatoid arthritis and pernicious anemia is described. Meticulous physical examination excluded further systemic or cutaneous involvement. The immunological workup revealed a low CD4 cell percentage with T cells mostly composed of CD8 cells, a discrepancy between the high percentage of cumulative CD4 + CD8 cells and the measured CD3 proportions, very low NK cytotoxicity toward K562 cells, and almost negligible responses to PHA, Con A and PWM mitogens. The results point to severe T and NK cell functional defects. The pathogenetic significance of these data is discussed. PMID- 8408924 TI - Cactus granuloma of the skin. AB - A case of cactus granuloma due to the thorns of Opuntia bieglovii Engelman is reported. A 24-year-old Japanese man contacted the thorns of this cactus in Palm Springs, California, U.S.A., and, three months later, numerous, slightly elevated papules 2 to 4 mm in diameter appeared on the dorsum of his right hand and fingers. Histopathological examination revealed many small granulomas surrounding the bristle fragments throughout the dermis. The granulomas consisted of epithelioid cells, Langhans type and foreign body type giant cells. The fragments of the barbed bristles gave strongly positive reactions with PAS. Two hypothetical mechanisms, one allergic and the other non-allergic, are offered to explain this granuloma formation and are discussed with reference to the literature. PMID- 8408925 TI - Fournier's gangrene: report of a case associated with paralytic and mechanical ileus throughout the management of the gangrene. AB - A 65-year-old Japanese male, who was treated for Fournier's gangrene, developed an enlarged erythema over the right thigh and right lower quadrant. The area was surgically debrided, and he was given antibiotics. However, he complained of abdominal swelling with a metallic bowel sound, pain, and vomiting and was then treated for paralytic ileus. Although his symptoms initially improved, he complained again of the same symptoms and underwent surgery for mechanical ileus occurring at the site of a surgical scar from an appendectomy 43 years earlier. This is a very rare case of Fournier's gangrene which caused mechanical ileus of the small intestine and adherence to a peritoneal scare after paralytic ileus due to inflammation of the abdominal fascia following scrotal gangrene. PMID- 8408926 TI - A case of superficial spreading melanoma in situ 2.5 mm in diameter. AB - A 60-year-old woman with superficial spreading melanoma in situ, measuring 2.5 mm in diameter, was examined. She had noticed a very small pigmented lesion 1.2 mm in diameter on her left lower leg in April of 1989. By April of 1990, it had grown to 2.5 mm in diameter. Its edge was irregular, and its color was variegated black to brown. Skin surface markings had disappeared in the center portion. Histopathologically, the lesion was asymmetrical. Atypical large cells nested in the lower epidermis and were scattered singly in the mid and upper epidermis, as seen in Paget's disease. At the periphery of the lesion, single large tumor cells were scattered in the mid epidermis. The tumor cells reacted to monoclonal anti melanoma antibody HMB-45. PMID- 8408927 TI - Relapsing syphilis. AB - Although penicillin still remains highly effective for syphilis, concerns have been raised that current regimens may be inadequate for the treatment of early syphilis. We report the failure of benzathine penicillin in the treatment of secondary syphilis. Extensive tracing of sexual contact history failed to identify sources of possible reinfection; therefore, we believe that this case represents a treatment failure and an ensuing relapse of secondary syphilis. PMID- 8408928 TI - Cytokine production of peripheral blood mononuclear cells in a dermatophytosis patient in response to stimulation with trichophytin. AB - Interferon-gamma (IFN-gamma), interleukin 2 (IL-2) and granulocyte/macrophage colony-stimulating factor (GM-CSF) were detected in the culture supernatant after 72 hours incubation with trichophytin in the peripheral blood mononuclear cells (PBMC) obtained from a patient who had a dermatophyte infection. These findings indicate that this patient has peripheral T-lymphocytes that produced IFN-gamma, IL-2 and GM-CSF, which may play roles in the development of delayed-type hypersensitivity (DTH) reaction in the skin. PMID- 8408929 TI - Long-term results of intralesional interferon alpha-2B in discoid lupus erythematosus. PMID- 8408930 TI - Blister formation over a soft fibroma of the nipple. AB - We report a case of soft fibroma on the nipple which was accompanied by a blister. Histologically, a subepidermal blister overlying the tumor and the degeneration of the lower part of the epidermis were observed. The causes of blister formation are briefly discussed. PMID- 8408931 TI - A case of acquired lymphangioma of the vulva. AB - A 46-year-old woman developed multiple papules on the right labium majus. A histological examination revealed acanthotic epidermis and dilated vessels in the papillary dermis. The clinical and histological features were compatible with those of lymphangiectasis or acquired lymphangioma of the vulva, which occurs after surgery or irradiation for cervical cancer. This patient, however, had no such past history. Acquired lymphangioma of the vulva arising without obvious causes seems to be unusual. PMID- 8408932 TI - Neurological intensive care in children. PMID- 8408933 TI - Respiratory response to salbutamol (albuterol) in ventilator-dependent infants with chronic lung disease: pressurized aerosol delivery versus intravenous injection. AB - OBJECTIVE: To compare the effects of intravenously injected with inhaled salbutamol in ventilator dependent infants with chronic lung disease (CLD). DESIGN: Prospective randomized study which each patient served as his/her own control. SETTING: Multidisciplinary neonatal and pediatric ICU. PATIENTS: 8 ventilator dependent premature infants with CLD. INTERVENTIONS: Salbutamol, 10 micrograms/kg was given intravenously, and 10-19 h later, twice 100 micrograms as pressurized aerosol, or vice versa, sequence randomized. The pressurized aerosol was delivered by a metered dose inhaler into a newly developed aerosol holding chamber, integrated into the inspiratory limb of the patient circuit. Respiratory system mechanics were assessed by the single breath occlusion method before and 10 and 60 min after drug administration. MEASUREMENTS AND RESULTS: Compliance improved significantly after intravenous injection (0.48 +/- 0.18 to 0.67 +/- 0.16, p < 0.01 and 0.59 +/- 0.23 ml/cmH2O/kg, NS, (mean +/- 1 SD) and after inhalation (0.46 +/- 0.19 to 0.64 +/- 0.32, p < 0.01 and 0.56 +/- 0.31 ml/cmH2O/kg, NS). Resistance decreased after iv. use (0.38 +/- 0.17 to 0.25 +/- 0.11, p < 0.001 and 0.25 +/- 0.10 cmH2O/ml/s, NS) and after inhalation (0.35 +/- 0.12 to 0.27 +/- 0.09, p < 0.01 and 0.28 +/- 0.12 cmH2O/ml/s, NS). Heart rate increased significantly after both routes of application, whereas mean arterial pressure, respirator settings, FIO2, transcutaneous SO2 and capillary PCO2 did not change. CONCLUSIONS: Inhaled and intravenous salbutamol improves pulmonary mechanics to the same extent with comparable side effects, and may therefore be used to facilitate weaning from respirators. PMID- 8408934 TI - Incidence and risk factors of pneumonia acquired in intensive care units. Results from a multicenter prospective study on 996 patients. European Cooperative Group on Nosocomial Pneumonia. AB - OBJECTIVE: To estimate the incidence of pneumonia acquired in the intensive care unit (ICU), and to define risk factors for developing such an event. DESIGN: European prospective survey, in which all patients admitted to the participating ICU from January, 17 to 23, 1990, were followed until ICU discharge. SETTING: 107 general ICUs from 18 countries. PATIENTS: Of 1078 admitted to the ICUs, 996 patients without pneumonia at admission were studied. MEASUREMENTS: Pneumonia was diagnosed by the staff physician on the basis of clinical, radiological and microbiological criteria, secondly validated by an expert committee who reviewed all the forms and even recontacted ICU physicians. Crude incidence and time to occurrence of pneumonia were estimated, then both used as end-points for prognosis analysis. RESULTS: 89 pneumoniae were observed: crude incidence was estimated at 8.9% 7-day and 14-day pneumonia rates at 15.8% and 23.4%, respectively. The risk of developing pneumonia increased when either coma, trauma, respiratory support, Apache II > 16 and/or imparied air-way reflexes were present at ICU admission. To predict time to occurrence of pneumonia, only two variables remained significant: the presence of impaired airway reflexes at admission and the use of mechanical ventilation during ICU course. CONCLUSIONS: The role of the injury to the respiratory system-with the subsequent need for respiratory support--appears central in determining the risk to acquire pneumonia in ICU. In the future, the predictive value of severity scores during ICU course should be otherwise assessed. PMID- 8408935 TI - Importance of pre-existing co-morbidities for prognosis of septicemia in critically ill patients. AB - OBJECTIVE: To determine admission characteristics associated with the outcome of septicemia in critically ill patients and more specifically assess the prognostic value of pre-existing co-morbidities. DESIGN: 5 year-retrospective cohort study. SETTING: Surgical Intensive Care Unit (ICU-20 beds) in a 1600 bed-tertiary care center. PATIENTS: Among 5457 patients admitted to the ICU between 1984 and 1988, 176 (3.2%) met prospectively-defined criteria for blood culture-proven septicemia (8.77 per 1000 patient-days). Overall septicemic patients had a 5-fold increased risk of death compared to non-septicemic patients (relative risk 5.03, 95% confidence intervals 4.17-6.07, p < 0.0001), and this estimate persisted after stratification according to age, sex, primary diagnosis and conditions of admission to the ICU (emergency/elective). RESULTS: Prognostic factors recorded on admission to ICU that were associated with mortality from septicemia among 173 patients were older age, higher admission Apache II score, gastrointestinal surgery, ultimately and rapidly fatal diseases and the number of co-morbidities in addition to the principal diagnosis (active smoking, alcohol abuse, non-cured malignancy, diabetes mellitus, splenectomy, recent antibiotic therapy, major surgery, or major cardiac event). In the multivariate analysis with logistic regression procedures, Apache II and co-morbidities were identified as the two independent predictors of mortality. CONCLUSIONS: Pre-existing co-morbidities assessed at the admission to the ICU significantly improved the prediction of mortality from septicemia compared to Apache II score alone. PMID- 8408936 TI - Epidemiological impact of prolonged systematic use of topical SDD on bacterial colonization of the tracheobronchial tree and antibiotic resistance. A three year study. AB - OBJECTIVE: to evaluate the effect of the prolonged systematic use of topical SDD (tobramycin 80 mg, polymyxin E 100 mg, amphotericin B 500 mg) on ICU ecology as expressed by changes in tracheal colonization and bacterial resistances. DESIGN: Prospective microbiological survey. SETTING: Polyvalent ICU of a 2000 beds general hospital. PATIENTS: Data concerning bacterial strains isolated from the tracheo-bronchial aspirates of all the patients admitted to a polyvalent ICU over 3 consecutive periods of 12 months ('88, '89, '90) were prospectively entered in a database and subsequently analyzed. During a 3-year period 502 patients required artificial ventilation for more than 72 h and 332 of them ('89 and '90) were treated with SDD. All samples collected within 72 h from ICU admission were excluded as well as duplicate samples from the same patients. INTERVENTION: All the patients admitted to the ICU in '89 and '90 and submitted to artificial ventilation for at least 24 h were routinely treated with topical SDD without i.v. antibiotic prophylaxis; in '88 SDD was not employed. MEASUREMENTS AND RESULTS: Criteria for collecting sputum samples and microbiological procedures remained unchanged throughout the study-time. Positive sputum were significantly less in '89 (80.8% versus 92.3% p < 0.001) and this was due to a very sharp decrease in the isolation of Gram-negative strains from 43-28% (-64% p < 0.0001) involving both: Enterobacteriaceae (-45%) and Pseudomonaceae (-77%). In 1990; however, a new increase in Gram negative was observed, although the overall amount of Gram-negative was still 49% lower in '90 if compared to '88 (p < 0.0001). A dramatic increase in Pseudomonas isolation was the only factor responsible for the "rebound" observed. An increasing percentage of Pseudomonas developed a resistance towards tobramycin and only 45% of Pseudomonas strains turned out to be sensible to tobramycin in '90 against 79% in '88. A similar trend was registered for all aminoglycosides with the exception of amikacin. Gram positive colonizations tended to increase (+63%) (p < 0.0001) and this was mainly due to Coagulase negative Staphylococci (+290% p < 0.0001) and S. pneumoniae, whereas S. aureus isolations decreased (-18%) but not significantly. CONCLUSIONS: Our data suggest that the prolonged use of SDD is associated with dramatic changes in ICU ecology: the incidence of Gram negative colonization is significantly diminished by SDD whereas Gram positive tend to increase. Pseudomonas developed an increasing resistance towards tobramycin one of the components of the SDD formula we used. PMID- 8408937 TI - Is penicillin G an adequate initial treatment for aspiration pneumonia? A prospective evaluation using a protected specimen brush and quantitative cultures. AB - OBJECTIVE: To evaluate the bacteriology of early aspiration pneumonia using a protected specimen brush and quantitative culture techniques, and whether penicillin G is adequate as initial treatment pending culture results. PATIENTS AND METHODS: 52 patients (of which 45 required mechanical ventilation) meeting usual clinical criteria for aspiration pneumonia were prospectively included. On admission, patients were given intravenous penicillin G and a protected specimen brush was performed < or = 48 h after. RESULTS: Cultures of the brush were negative (< 10(3) CFU/ml) in 33 patients (1 had blood cultures positive with S. pneumoniae) and positive (> or = 10(3) CFU/ml) for S. pneumoniae in 2 patients. Seventeen patients had a positive culture (> or = 10(3) CFU/ml) for at least one penicillin G resistant microorganism, with a total of 20 organisms (S. aureus: 6; H. influenzae: 2; Enterobacteriaceae: 8; P. aeruginosa: 3; C. albicans: 1). In 4 of these patients, a penicillin-sensitive pathogen was also recovered in significant concentrations (S. pneumoniae: 2; Streptococcus sp.: 2). These 17 patients with a resistant pathogen did not differ from the 35 other patients with respect to need for ventilatory support and mortality rate. By contrast, they were older (61.1 +/- 21.9 vs. 42.9 +/- 18.8 years; p < 0.005) and required longer mechanical ventilation (6.1 +/- 4.6 vs. 3.5 +/- 2.7 days; p < 0.03) and hospitalization (11.2 +/- 8.8 vs. 6.7 +/- 4.7 days; p < 0.02). Of 17 patients 12 with penicillin G resistant organisms versus 0/35 without, were in-hospital patients and/or had a digestive disorder (p < 0.001). CONCLUSION: The broad range of offending organisms seen in early aspiration pneumonia precludes use of any single empiric regimen, making protected specimen brush mandatory in many patients. Nevertheless, the involvement of S.pneumoniae in a notable proportion of our patients suggests that routine penicillin prophylaxis after early aspiration (at least in most patients with community-acquired aspiration) is warranted given the potential severity of pneumococcal sepsis in such patients. PMID- 8408938 TI - Resistance to methicillin and virulence of Staphylococcus aureus strains in bacteriemic cancer patients. AB - OBJECTIVE: To estimate the relation between resistance to methicillin and virulence in strains of Staph. aureus by estimating the survival at day 30. DESIGN: Case control analysis. SETTING: Inpatients at a major Cancer Referral Center. PATIENTS: 21 patients with methicillin-resistant Staph. aureus (MRSA) bactaeremia and 45 patients with methicillin-susceptible Staph. aureus (MSSA) bactaeremia, all treated with vancomycin. MEASUREMENTS AND MAIN RESULTS: Fewer MRSA than MSSA patients were neutropenic (p = 0.04), but more MRSA patients were hospitalized in intensive care units (p = 0.03), had had surgical episodes (p = 0.06). MRSA patients also had more microbiological samples cultured (p = 0.007). The outcome of the bacteriemic episodes in 14 MRSA and 14 MSSA patients matched for these four factors showed that blood cultures from MRSA patients remained positive significantly longer (p = 0.04), but that survival and length of hospital stay were not significantly different. CONCLUSIONS: These results do not indicate that methicillin-resistance in Staph. aureus is associated with increased virulence. PMID- 8408939 TI - Continuous veno-venous haemofiltration following cardio-pulmonary bypass. Indications and outcome in 35 patients. AB - OBJECTIVE: To study the impact of continuous veno-venous haemofiltration on survival in patients with acute renal failure (ARF) following cardio-pulmonary bypass (CPB) surgery. DESIGN: A retrospective study of all patients requiring haemofiltration after CPB over a 2 year period. SETTING: A 20 bedded, adult cardothoracic intensive care unit in a postgraduate teaching hospital. PATIENTS: 35 patients (26 male, age range 24-74 years) required haemofiltration (2.7% of the total number of patients undergoing CPB). MAIN RESULTS: Cardiovascular failure post CPB was the commonest causes of ARF (n = 16). Indications for haemofiltration were uremia (21), oligo-anuria (11), volume overload (2) and hyperkalaemia (1). Mean time from CPB to the initiation of haemofiltration was 8 days (range 0-15 days). Mean urea was 30 mmol/l and creatinine 362 mumol/l immediately prior to treatment. Urea was well-controlled in all patients, although 2 needed haemodiafiltration. Twenty-six patients died during their admission to the ICU (74% mortality). A further 3 patients died during their hospital admission, following discharge from ICU. Outcome was particularly poor in patients with cardiovascular failure following CPB (16 cases, 0 survivors). Survivors tended to commence filtration earlier (mean of 4 vs 7 days for non survivors) and required treatment for a mean period of 8 days (range 1-26 days). Survival was determined by the number of failed organ systems at the start of haemofiltration. Thus, 100% of patients with single system failure survived, compared to only 10% with 3 or more system failure. CONCLUSIONS: Despite the theoretical advantages of haemofiltration and the effective control of uraemia the mortality associated with ARF following CPB remains high and is probably determined by the number of failed organs systems. PMID- 8408940 TI - Three different mask physiotherapy regimens for prevention of post-operative pulmonary complications after heart and pulmonary surgery. AB - OBJECTIVE: An investigation into the incidence of post-operative complications after thoracic surgery with 3 different physiotherapy masks. DESIGN: A prospective, consecutive, randomized comparison. SETTING: Department of Thoracic and Heart Surgery at a University Hospital. The treatments were performed by experienced and specially trained physiotherapists. PATIENTS: 160 patients were evaluated. 60 patients undergoing heart surgery, 59 patients having pulmonary resection, and 41 patients with exploratory thoracotomy. INTERVENTIONS: In each operative category the patients were treated with one of three face mask systems used in addition to routine chest physiotherapy. These were either continuous positive airway pressure (CPAP), positive expiratory pressure (PEP), or inspiratory resistance - positive expiratory pressure (IR-PEP). MEASUREMENTS AND RESULTS: Post-operative pulmonary complications were assessed by forced vital capacity (FVC), arterial oxygen tension (PaO2), and chest X-ray examination, all measured pre-operatively and on the fourth and ninth post-operative day. The patients filled in a questionnaire expressing their opinion about their mask treatment. There was an equal decrease in FVC, FVC%, and PaO2, and equal frequency of atelectasis in the 3 mask treatments. More patients with the PEP mask favoured their system than did those with the other 2 systems. CONCLUSION: There was no statistically significant difference between the treatments: continuous positive airway pressure (CPAP), positive expiratory pressure (PEP), and inspiratory resistance - positive expiratory pressure (IR-PEP) on post operative complications. Any of the three treatments may be used as supplement to standard chest physiotherapy. PMID- 8408941 TI - Propofol emulsion reduces proliferative responses of lymphocytes from intensive care patients. AB - OBJECTIVE: To test propofol lipid emulsion formulation for its immunosuppressive effects. DESIGN: Propofol lipid emulsion and the emulsion alone were tested at increasing concentrations and compared to initial values and between each other. Propofol alone could not be tested due to its insolubility into the culture medium. PATIENTS AND PARTICIPANTS: Lymphocytes from 12 surgical intensive care (ICU) patients (median APACHE score 16 and median TISS score 28) and 12 healthy volunteers. MEASUREMENTS: Phytohaemagglutinin-, concanavalin A- and pokeweed mitogen-induced lymphocyte proliferative responses were measured in the presence of increasing concentrations of propofol lipid emulsion formulation or the lipid emulsion. RESULTS: Lymphocyte proliferative responses from ICU patients were in general on a lower level than in the volunteers. The propofol lipid emulsion formulation (Diprivan) decreased pokeweed mitogen-induced proliferative responses of lymphocytes from ICU patients at propofol concentrations found in the circulation (1-10 micrograms/ml) and the lipid emulsion alon at 100 micrograms/ml triglyceride concentrations while the other mitogen-induced responses were not affected. No changes were observed in the mitogen-induced responses of lymphocytes from healthy volunteers. CONCLUSIONS: Propofol emulsion formulation decreased in surgical intensive care patients pokeweed mitogen-induced lymphocytic responses in vitro at clinically found concentrations, indicating the need for further studies to test B-lymphocyte functions and T-B-lymphocyte co operation during propofol lipid emulsion administration. (ICU) patients is widespread because of its good control of sedation. Propofol is currently administered in fat emulsion which is considered immunosuppressive during bolus injection or rapid infusion. Therefore, effects of a propofol fat emulsion formulation on proliferative responses of lymphocytes were studied in blood samples obtained from healthy volunteers and ICU patients known to be immunosuppressed. PMID- 8408942 TI - The role of blood microfilters in clinical practice. PMID- 8408943 TI - Validation of diagnostic tests. PMID- 8408944 TI - Propofol induces bronchodilation in a patient mechanically ventilated for status asthmaticus. PMID- 8408945 TI - Expressed emotion, social skill, and response to negative affect in schizophrenia. AB - The social skills and social perception of schizophrenia patients in response to negative affect was examined as a function of family expressed emotion (EE). Patients participated in a role-play test, a social perception test, and a problem-solving discussion with a family member and were assessed on several measures of symptomatology. EE of family members was evaluated with the Camberwell Family Interview. On the role-play test, patients with less critical relatives became more assertive in response to increased negative affect from a confederate portraying either a family member or friend, but patients with highly critical relatives did not. Patients with highly critical relatives were also less assertive when confronted with negative affect from a confederate portraying a family member rather than a friend. The behaviors of both relatives and patients during a family problem-solving interaction were related to the EE dimensions of criticism, emotional overinvolvement, and warmth. Patient gender was also related to family problem solving but was independent of EE. Patient ratings of affect on a videotaped social perception task were not related to family EE, and there were few differences in psychopathology between patients with high and low EE relatives. The results support the validity of the EE construct as an index of relatives' affective behavior and suggest that patients' social skills, such as assertiveness, may mediate negative affective exchanges in their families. PMID- 8408946 TI - Cerebral asymmetry and hypnosis: a signal-detection analysis of divided visual field stimulation. AB - Brightness discriminations in a divided-field paradigm were examined with a signal-detection procedure in three sessions, the second with hypnosis. Practiced, hypnotically susceptible Ss were subdivided into high- (n = 6) and medium- (n = 5) susceptible groups on the basis of a susceptibility scale monitored throughout the hypnosis session. High-susceptible Ss showed increases in d' in the left visual field (right hemisphere) with hypnosis, whereas medium susceptible Ss showed bilateral enhancements. Beta remained invariant in both groups across three sessions but was higher in the left visual field. The results provide evidence of altered brain function with hypnosis and an association of focal right hemispheric changes with high susceptibility and, through the invariance of beta, fail to support the attribution of perceptual changes to attitudinal, nonstate factors. PMID- 8408947 TI - Differences between depressed and nondepressed individuals in the recognition of and response to facial emotional cues. AB - The present study investigated the recognition of, and responses to, facial expressions of emotion. Participants were all women and consisted of the following groups: (a) Sixteen depressed college students; (b) 16 nondepressed college students; (c) 16 depressed psychiatric patients; and (d) 11 nondepressed psychiatric patients. Results suggest that both depressed groups, relative to the nondepressed college group, made more errors in recognizing the facial expressions and reported more freezing or tensing; higher fear and depression reactions; and less comfort with their own emotional reactions to these expressions and a stronger desire to change these reactions. Few differences were found between the depressed psychiatric patients and the psychiatric control subjects. It is concluded that inappropriate reactions to others' emotions may maintain or increase depression. PMID- 8408948 TI - Early language and intelligence development and their relationship to future criminal behavior. AB - The relationship between intelligence, measured regularly from the ages of 3 to 17 years, and registered criminality was investigated for boys (N = 122) in a birth-to-maturity study. Significant negative correlations appeared at several ages, even for intelligence assessed as early as at the age of 3. The hypothesis was advanced that the early language development of the boys would be negatively associated with future criminality. Information on language development, obtained by applying the Brunet-Lezine psychomotor developmental test for infants, substantiated this hypothesis. Significant correlations with registered criminality appeared for language development at 6, 18, and 24 months. Further support for the hypothesized link was provided by psychologists' ratings of children's verbal behavior and by maternal reports of their child's speech at the ages 3 to 5. The role of early language retardation in contributing to later criminality is discussed. PMID- 8408949 TI - Altering a dominant response: performance of psychopaths and low-socialization college students on a cued reaction time task. AB - The passive avoidance learning deficits of disinhibited Ss have been attributed to their difficulty inhibiting dominant responses. To date, evidence for this hypothesis has been derived from complex tasks. In two experiments, a cued reaction time task requiring no learning or memory was used to evaluate the degree to which groups of disinhibited Ss inhibit simple dominant responses. Disinhibited groups were incarcerated psychopaths identified with Hare's (1985) Psychopathy Checklist and undergraduate males who scored low on the Socialization Scale. Both disinhibited groups committed more errors than controls on trials containing misleading cues, but in both samples, findings were limited to trials in which Ss expected to make right-hand responses. Although alternative interpretations are possible, these data are consistent with the proposal that disinhibited individuals are less likely to inhibit well-established dominant responses. PMID- 8408950 TI - Lack of synchronized seasonal variation in the intensity of psychological problems. AB - Temporal variation in the subjective intensity of psychological problems identified by 40 psychotherapy clients (most diagnosed as depressed) was investigated longitudinally. Each client rated the intensity of 10 individualized problems 3 times per week (on Tuesdays, Thursdays, and Sundays) for an average of 6 months. Ratings from all clients were spread over 3 years. When scores were statistically adjusted for client differences and average rate of improvement, no significant variation among months was found, regardless of problem content or client gender. Standard assessment measures administered 4 times to each client also failed to show systematic seasonal variation. PMID- 8408951 TI - Behavioral and emotional problems among Thai and American adolescents: parent reports for ages 12-16. AB - We studied adolescents' behavioral and emotional problems in the United States and in Thailand, a Buddhist country in which, reportedly, aggression is discouraged and self-control, emotional restraint, and social inhibition are encouraged. Standardized parent reports on 118 problems revealed 45 Thai-U.S. differences. Thai adolescents were reported to show more overcontrolled problems (e.g., shyness, compulsivity, inhibition of talking, fearfulness, and constipation) than American adolescents (p < .0001). The two groups did not differ reliably in total undercontrolled problems, but Americans showed higher levels of direct, overt, and interpersonally aggressive undercontrol (e.g., fighting and bullying), whereas Thais showed more indirect and subtle undercontrol that was not interpersonally aggressive (e.g., sulking and sullenness). The findings suggest that different cultures may be linked to different styles of adolescent problem behavior. PMID- 8408952 TI - Stress reaction of school-age children to the bombardment by SCUD missiles. AB - Stress reactions of 5th-, 7th-, and 10th-grade children (N = 492) exposed to missile attacks during the Persian Gulf War were examined a month after the war by a questionnaire that assessed level of exposure to trauma and psychological symptoms. Higher stress responses were obtained in areas hit and were influenced by proximity to sites or individuals involved in actual damage. Gender, age, and region interacted such that 5th-grade boys reported the highest stress reactions regardless of region, whereas 5th-grade girls reported the highest stress responses only in regions hit. Gender, age, and objective and subjective stress correctly identified 75% of the children as potential clinical or nonclinical candidates. The possible mediating coping responses and applications for high risk groups are discussed. PMID- 8408953 TI - Control-related beliefs and self-reported depressive symptoms in late childhood. AB - Research thus far links depressive symptoms in children to one type of control related belief: low levels of perceived personal competence. However, child research, unlike adult research, has not supported a linkage between depressive symptoms and another theoretically important control-related belief: perceived noncontingency of outcomes. Here we reexamined the issue, adjusting for limitations in previous methodology by using (a) psychometrically stronger measures of control beliefs, and (b) a general population sample rather than children being treated in mental health clinics. In contrast to previous results, we found that both perceived incompetence and perceived noncontingency were strongly related to children's depression, together accounting for 40% of the variance in Child Depression Inventory scores. We also found, as in previous research, that depressive symptoms were correlated with uncertainty as to the causes of outcomes, especially successes. The findings suggest that children may be susceptible to both "personal helplessness" and "universal helplessness" forms of depression. PMID- 8408954 TI - Detrimental effects of kin support networks on the course of depression. AB - On the basis of preliminary findings that, for recovered depressed patients, a good family support network is associated with a subsequent increase in depressive symptoms, a prospective study is presented which examines the specific conditions under which this counterintuitive effect is present. A sample of 168 depressed psychiatric inpatients was assessed 1 and 7 months after discharge from hospital regarding, among other variables, depressive symptomatology, diagnostic status, and the composition and supportive functions of their social networks. The detrimental effect was restricted to female patients who were recovered at discharge and who were homemakers, and it involved supportive relatives, in particular relatives providing close psychological and emotional support in crises. Possible psychological and psychosocial mediators as well as implications for research and intervention are discussed. PMID- 8408955 TI - Voices of fear and anxiety and sadness and depression: the effects of speech rate and loudness on fear and anxiety and sadness and depression. AB - Two studies investigated the role of expressive vocal behavior (specifically, speech rate and loudness) in fear and anxiety and in sadness and depression. In the first study, participants spoke about personally experienced fear and anxiety arousing and neutral events using 3 different voice styles: fast and loud, normal, and slow and soft. In the second study, participants spoke about personally experienced sad or depressing and neutral events using the same 3 voice styles. In both studies, the participants' highest levels of subjective affective and cardiovascular (CV) arousal occurred when they spoke about the emotional events in a mood-congruent voice style: fast and loud in the case of fear and anxiety, and slow and soft in the case of sadness or depression. Mood incongruent voice styles canceled the heightened levels of CV arousal normally associated with these negative emotions. The voice-style manipulation had no significant effect on the participants' levels of CV arousal during the neutral discussions. PMID- 8408956 TI - Personality and behavioral vulnerabilities associated with risk status for eating disorders in adolescent girls. AB - This article presents first-year cross-sectional findings from a study of the development of eating disorders. Adolescent female (N = 937) 7th through 10th graders completed measures that included information on personality, self concept, eating patterns, and attitudes. A risk status score was calculated on the basis of comprehensive information regarding DSM-III-R eating disorders criteria and other weight and attitudinal data. All personality measures showed significant differences according to risk, based on subject classification into high, moderate, and mild risk status and comparison groups. Early puberty was not associated with increased risk. The strongest predictor variables for risk were body dissatisfaction, negative emotionality, and lack of interoceptive awareness. The possible diathesis of personality including temperamental factors in the later development of an eating disorder is discussed. PMID- 8408957 TI - Structural cerebral pathology in schizophrenia: regional or diffuse? AB - Is brain pathology in schizophrenia topographically distinct? If so, are the putative regional changes unique to the disorder? To address these questions, 56 chronic schizophrenic Ss were compared with 16 psychiatric control Ss with mood disorders and with 31 healthy volunteers on multiple-volume measures of regional cerebral atrophy obtained with computed tomography. Generalized cortical and subcortical enlargement of spaces filled with cerebrospinal fluid sparing only the occipitoparietal cortex was found in the schizophrenic Ss compared with normal control Ss. Statistically significant differences in the extent of perisylvian atrophy were noted between schizophrenic Ss and patients with mood disorders: Schizophrenic Ss evidenced greater dilation of perisylvian fissures and sulci. The implications of the results for future research and for recent theories on the etiology of schizophrenia are discussed. PMID- 8408958 TI - Emotional learning, hedonic change, and the startle probe. AB - A multiple-response analysis of aversive learning was conducted in human subjects. For each subject, two pictorial stimuli were presented--one paired with electric shock. After training, the magnitude of the acoustic startle eyeblink reflex elicited in the context of the shocked picture increased dramatically and was significantly larger than for reflexes elicited during the nonshocked stimulus. Five different picture contents were tested in separate groups: Reflex potentiation was larger for pictures rated as pleasant than pictures rated as unpleasant. Conditioned responses were also evident for skin conductance, heart rate, and affective judgments. Different systems reflected different aspects of the acquired fear response: Conductance change covaried with arousal, and startle probe magnitude varied with affective valence (pleasure). The neurophysiological implications of the data are elucidated, and parallels drawn between animal and human subjects findings. PMID- 8408959 TI - Psychophysiological consequences of unexplained arousal: a posthypnotic suggestion paradigm. AB - This experiment compared the emotional, cognitive, and physiological responses of Ss experiencing induced physiological arousal with and without awareness of the source of their arousal. Nine highly hypnotizable Ss and 9 nonhypnotizable controls were used in a within-subjects design. Each S received posthypnotic suggestions for arousal (increases in heart and respiration rate) with and without amnesia for its source in a two-phase procedure. Only the hypnotizable Ss were expected to differ between conditions. As predicted, for the hypnotizable Ss, unexplained arousal produced significant and dramatic effects when compared with explained arousal, including misattributions. These results are considered within a conceptual framework of the role of discontinuous experiences in the development of psychopathological symptoms in normal persons. PMID- 8408960 TI - The effect of stress on hedonic capacity. AB - Two experiments were conducted to examine the effect of stress on hedonic capacity. In Experiment 1, cadets in the U.S. Army Reserve Officer Training Corps reported experiencing less pleasure while watching amusing film clips after participating in a weekend of field training exercises than they did on a control day. In Experiment 2, college students reported experiencing less pleasure in their daily activities during final exam week than they did at the baseline assessment. Hedonic capacity was associated with positive affect but not with negative affect or reported stress levels. The deleterious effect of stress on hedonic capacity was particularly strong for subjects with family histories of depression. PMID- 8408961 TI - Interpersonal deviance and consequent social impact in hypothetically schizophrenia-prone men. AB - Interpersonal deviance is central to the theory of and research on schizotypal psychopathology. The present study investigated interpersonal deviance and its corresponding impact among hypothetically schizotypic, or schizophrenia-prone, men, defined by high scores on the Perceptual Aberration-Magical Ideation (Per Mag) Scale. In a videotaped interview, high-scoring Ss relative to control Ss were rated as more odd (p < .001) and more avoidant (p < .05) in their interview behavior and made the interviewers feel more anxious (p < .05), more angry (p < .05), and less interested (p < .05). Other analyses revealed that oddness was the strongest discriminating variable and that this behavior could not be accounted for by social anxiety or lack of interest. These results provide further construct validation for the Per-Mag scale and suggest that interpersonal factors may influence the eventual adjustment of high-scoring individuals. PMID- 8408962 TI - Alcohol consumption and sexual activity as reported with a diary technique. AB - Although alcohol is commonly viewed as a sexual disinhibitor, recent research has suggested that alcohol consumption does not consistently lead to increased sexual activity. Nonexperimental work in this area has commonly used correlational procedures that do not control for individual difference variables that may contribute to a drinking-sex relationship. This study examined the relationship of alcohol consumption to sexual behavior by way of within-subjects analyses of data from 99 men and women who kept daily diaries of drinking and sexual events over a 10-week period. Alcohol consumption was associated with a general attenuation of sexual activity, with no effects on the occurrence of sexual behaviors that are risky in terms of AIDS transmission. These data suggest that sexual distribution and lapses in sexual judgment are not necessarily common consequences of alcohol consumption. PMID- 8408963 TI - Emotional and behavioral responses to aversive interpersonal behaviors. AB - In a conceptual replication of A. Biglan, J. Rothlind, H. Hops, and L. Sherman (1989), 288 subjects rated their emotional and behavioral responses to aggressive, distressed, and neutral behaviors. The stimulus behaviors occurred in the context of casual relationships and were portrayed by male and female actors. Consistent with Biglan et al., we found that aggressive behaviors evoked angry emotions and negative behavioral responses; distressed behaviors evoked both angry and concerned emotions and supportive behavioral responses. However, in contrast to the original results, we also found evidence of negative (primarily avoidant) behavioral responses to distressed behavior. PMID- 8408964 TI - Training Japanese listeners to identify English /r/ and /l/. II: The role of phonetic environment and talker variability in learning new perceptual categories. AB - Two experiments were carried out to extend Logan et al.'s recent study [J. S. Logan, S. E. Lively, and D. B. Pisoni, J. Acoust. Soc. Am. 89, 874-886 (1991)] on training Japanese listeners to identify English /r/ and /l/. Subjects in experiment 1 were trained in an identification task with multiple talkers who produced English words containing the /r/-/l/ contrast in initial singleton, initial consonant clusters, and intervocalic positions. Moderate, but significant, increases in accuracy and decreases in response latency were observed between pretest and posttest and during training sessions. Subjects also generalized to new words produced by a familiar talker and novel words produced by an unfamiliar talker. In experiment 2, a new group of subjects was trained with tokens from a single talker who produced words containing the /r/-/l/ contrast in five phonetic environments. Although subjects improved during training and showed increases in pretest-posttest performance, they failed to generalize to tokens produced by a new talker. The results of the present experiments suggest that variability plays an important role in perceptual learning and robust category formation. During training, listeners develop talker specific, context-dependent representations for new phonetic categories by selectively shifting attention toward the contrastive dimensions of the non native phonetic categories. Phonotactic constraints in the native language, similarity of the new contrast to distinctions in the native language, and the distinctiveness of contrastive cues all appear to mediate category acquisition. PMID- 8408965 TI - A cross-linguistic investigation of locus equations as a phonetic descriptor for place of articulation. AB - A previous study [H. Sussman, H. McCaffrey, and S. Matthews, J. Acoust. Soc. Am. 90, 1309-1325 (1991)] of American English CV coarticulation showed a remarkably linear relationship between onset frequencies of F2 transitions, plotted on the y axis, in relation to the F2 midvowel "target" frequencies, plotted on the x axis, for CVC tokens with initial [b d g] and ten medial vowel contexts. Slope and y intercept values of regression functions fit to these scatterplots ("locus equations") were shown to serve as statistically powerful phonetic descriptors of place of articulation. The present study extends the locus equation metric to three additional languages--Thai, Cairene Arabic, and Urdu--having both two and four place contrasts for syllable-initial voiced stops. A total of 14 speakers (Thai = 6, Arabic = 3, Urdu = 5) produced 1740 CVC tokens that were acoustically analyzed using MacSpeech Lab II. Strong linear regression relationships were found for every stop category across all speakers. Slopes and y intercepts systemically varied as a function of place of articulation. Cross-language comparisons of stop place categories were performed but variability of slope and y intercept means tempered conclusions concerning the existence of CV "phonetic hot spots." PMID- 8408966 TI - Manipulations of the duration and relative onsets of two-tone forward maskers. AB - The effects of absolute and relative masker duration were investigated using one- and two-tone forward maskers. In the absolute-duration conditions, the duration (D) of a one-component on-frequency (M1; Dm1-alone) or off-frequency (M2; Dm2 alone) masker or the common duration of a two-component masker (Dm1 = Dm2) was varied. Masked threshold increased as duration increased in all three conditions. In the relative-duration conditions, the duration of one masker component was fixed at a relatively long value and the duration of the other component was varied (Dm1 > Dm2 or Dm2 > Dm1). In the Dm1 > Dm2 condition, masked threshold decreased as the duration of Dm2 was increased. In the Dm2 > Dm1 conditions, masked threshold increased and then decreased slightly as the duration of Dm1 was increased. Masked threshold was frequently lower in the M1+M2 than in the M1 alone conditions, indicating psychophysical suppression. Further, masked threshold in the Dm2 > Dm1 condition was always higher than in the corresponding absolute-duration condition (Dm1 = Dm2) having the same M1 duration, indicating masker enhancement. A two-stage neural model was proposed to account for these forward-masking data. PMID- 8408967 TI - A composite randomization procedure for measuring spectral shape discrimination. AB - In studies of auditory profile analysis [D. M. Green, Profile Analysis: Auditory Intensity Discrimination (Oxford U. P., New York, 1988)], the sounds are presented at random levels to discourage the listener from basing the discrimination on a difference in absolute level rather than on a difference in the shape of the spectrum. A difference in absolute level, however, can still provide an effective discrimination cue if the difference is comparable to the range of randomization. Using enormous ranges of random levels is not desirable, because it is distracting to normal listeners and may exceed the dynamic range of hearing for listeners with hearing loss. This article describes a new experimental procedure which permits the experimenter to greatly reduce the range of level randomization in roving-level tasks. PMID- 8408968 TI - Loudness adaptation at high frequencies. AB - Loudness adaptation was measured for pure tones at 4, 12, 14, and 16 kHz. In three experiments, a total of 87 young listeners judged--by the method of successive magnitude estimation--the loudness of the tones over a 6-min exposure period. Thresholds were measured by an adaptive 2IFC procedure. Although earlier measurements had shown that adaptation near threshold increases with frequency, these new data reveal that the increase is especially marked at higher sensation levels. Thus, at 40 dB SL, over a 6-min period loudness declined by 18% at 4 kHz and by 94% at 16 kHz. Moreover, the 4-kHz tone remained audible for all listeners throughout the 6-min exposure period whereas the 16-kHz tone became inaudible for two-thirds of them by the end of the exposure period. Listeners with relatively low thresholds (< 50 dB SPL) at 16 kHz showed much less adaptation at 14 kHz than at 16 kHz and much less than listeners with relatively high thresholds (> 50 dB SPL) at 16 kHz; this latter group showed strong adaptation at 14 kHz. The marked loudness adaptation of steady tones at very high frequencies and relatively high sensation levels is associated with a restricted spread of excitation in the auditory system resulting from the steep rise of the threshold curve at the highest audible frequencies. PMID- 8408969 TI - Stimulus features affecting psychophysical detection thresholds for electrical stimulation of the cochlea. II: Frequency and interpulse interval. AB - Psychophysical detection thresholds for electrical stimulation of the cochlea were measured in nonhuman primates (macaques) as part of a series of experiments exploring the stimulus features affecting detection. The monkeys were trained psychophysically using operant conditioning. One ear was treated with neomycin to destroy hair cells, and implanted with electrodes in the scala tympani and/or the cochlear wall. In experiment 1, detection thresholds were measured for trains of fixed-duration pulses and for sinusoids. For long-duration pulses (1 to 2 ms/phase), thresholds decreased as a function of frequency (pulse rate), reaching a minimum at a frequency between 125 and 210 pps, then increased as frequency was further increased. For shorter duration pulses, thresholds usually decreased monotonically as a function of frequency but sometimes showed a slight increase as a function of frequency near the highest frequencies tested. Typically slopes of the threshold versus frequency functions for fixed-duration pulses were equal to or less than slopes of threshold versus frequency functions for sinusoidal signals, where frequency and phase duration covaried. Additional observations on two of the cases were made in experiments 2 and 3. In experiment 2, thresholds for pairs of pulses were measured as a function of interpulse interval. Thresholds decreased as a function of interpulse interval up to intervals of 2 to 4 ms and then increased slightly. In experiment 3, thresholds were measured as a function of stimulus duration at two frequencies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8408970 TI - Contributions of comodulation masking release and temporal resolution to the speech-reception threshold masked by an interfering voice. AB - Two experiments are presented to explain the difference in speech-reception threshold (SRT) between conditions with a steady-state noise masker or an interfering voice. Literature shows for normal hearing a masking release of 6-8 dB with an interfering voice and a substantial reduction of this release with hearing impairment. In experiment I the possible role of comodulation masking release (CMR) is investigated by manipulating the comodulation in the interfering voice by the introduction of temporal shift among filter bands of various width. The spectral spread of masking from the manipulated interfering voice was controlled by interleaving the mutually shifted speech bands with 1/3-octave bands of noise. Although comodulation in the interfering speech appears to be very important for the low SRT, the contribution of across-frequency processing of masker fluctuations--commonly considered as the origin of CMR--is only 1.3 dB. In experiment II the level dependence of masking release with an interfering voice is investigated. The data fit in with the hypothesis by Festen and Plomp [J. Acoust. Soc. Am. 88, 1725-1736 (1990)] that the release from masking with an interfering voice is limited by forward masking. It appears that up to about 55 dBA the release from masking increases with level up to about 7 dB. Above 55 dBA the difference in SRT obtained with a noise masker or an interfering voice is constant due to the limited average modulation depth of speech. PMID- 8408971 TI - The relationship between sound transfer functions from free sound field to the eardrum and temporary threshold shift. AB - Three groups of 12 subjects were utilized for an experiment in which the temporary threshold shift due to a free-field exposure was measured and compared to the sound transfer functions from free-sound field to the subjects' eardrums. Subject selection was based upon their sound transfer functions and the requirement of normal hearing. The subjects were assigned to one of three groups depending on their dominantly sound transfer function frequency: Low, middle, or high. A probe tube with a miniature microphone was used for the measurement of the sound transfer functions in 1/3-oct bands. The measurements were performed in frequency bands from 0.2 to 20 kHz with one direction of sound incidence: 0 degree azimuth and 0 degree elevation. The subjects were exposed twice to a 2-kHz and twice to a 4-kHz narrow-band noise monotonically on four different occasions. Pre- and postexposure sweep Bekesy audiograms were recorded and the temporary threshold shift calculated as the difference between the two. The averaged temporary threshold shifts differed significantly among groups. PMID- 8408972 TI - Limited resolution of spectral contrast and hearing loss for speech in noise. AB - This paper examines the relations among the spectral contrast needed for speech intelligibility, hearing loss for speech in noise, and auditory filter bandwidth. Fifteen hearing-impaired listeners with relatively flat, mild-to-moderate sensorineural losses and eight normal-hearing listeners participated in the study. The spectral contrast needed for speech intelligibility was determined by reducing spectral contrast in the speech signal and measuring the reduction in contrast beyond which the speech-reception threshold (SRT) for sentences in noise increases. Reduction of spectral contrast was accomplished by smearing the envelope of the squared short-time fast Fourier transform by a convolution with a Gaussian-shaped filter, and overlapping additions to reconstruct a continuous signal. Auditory filter bandwidth was determined by estimating auditory filter shapes at center frequencies of 0.8, 1.6, and 3.2 kHz, using a notched-noise masking paradigm. The results show that limited resolution of spectral contrast is only loosely associated with hearing loss for speech in noise. Moreover, the correlations between the SRT for unsmeared speech and the auditory filter bandwidth at various frequencies were weak. PMID- 8408973 TI - Further evidence for an autonomous processing of pitch in auditory short-term memory. AB - In four related experiments, subjects had to discriminate between the presence or absence of a frequency difference between two pure tones separated by 4.3 s. The interference effects of other tones (I), inserted during the retention interval, were investigated. A previous study [C. Semal and L. Demany, J. Acoust. Soc. Am. 89, 2404-2410 (1991)] had shown that subjects' performance strongly depended on the pitch relations between the test tones and the I tones, but not on the spectral composition of the I tones. This suggested that the mnemonic system processing pitch is deaf to sharpness of timbre. In the present study, the I tones could differ or not from the test tones in intensity (by +/- 6 or 15 dB) or in amplitude envelope (periodic as well as aperiodic envelopes were used). It was found that such differences had very little effect on performance, suggesting that the mnemonic system processing pitch is deaf to loudness and to dynamic aspects of timbre. However, for I tones with a dense spectrum in the vicinity of the test tones' frequencies, widening the I tones' spectrum improved performance, probably because this spectral widening decreased the salience of the I tones' pitches. PMID- 8408974 TI - On the potential of fixed arrays for hearing aids. AB - Microphone arrays with fixed (time-invariant) weights are directed at enhancing a desired signal from one direction (straight ahead) while attenuating spatially distributed interference and reverberation. Using the theory of sensitivity constrained optimal beamforming [Cox et al., IEEE Trans. Acoust. Speech Sig. Process. ASSP-34, 393-398 (1986)], free-field arrays of head-sized extents were studied. The key parameters affecting array design and performance are the set of transfer functions from the target direction to each array microphone [H(f)] and the intermicrophone cross-spectral densities for isotropic noise [Szz(f)]. Design variables included the orientation of the array, the number, and [as motivated by Soede, Ph.D. thesis, Delft University of Technology (1990)] the directionality of the microphones within the array, and the complexity and robustness of the required processing. Performance was characterized by the broadband intelligibility-weighted directivity (gain against isotropic noise) and noise sensitivity (reflecting the array's sensitivity to uncorrelated noise, as well as device tolerances). For broadside orientation, a variety of arrays based on cardioid and hypercardioid microphones gave very similar performance. They can provide directivities of 7-8 dB with easily implemented weights (simple scalars). For endfire orientation, as Soede (1990) recognized, similar directivities result with weights based on analog gains and pure time delays. However, with weightings chosen independently for each frequency, directivities up to approximately 11 dB may be obtained, although the increased noise sensitivities of these arrays require practical evaluation. Because of sound diffraction, placement of arrays onto the head potentially impacts both their design and performance. In-situ measurements of H(f) and Szz(f) as well as simplified theoretical models are suggested to explore the optimization of head-mounted arrays. PMID- 8408975 TI - Measuring human cochlear traveling wave delay using distortion product emission phase responses. AB - A method is presented here in which cochlear traveling wave delays are estimated through the measurement of distortion product emission phase (DPE) responses. This method assumes that the site of generation of DPEs is at the f2 place. Eighteen adult female and 18 adult male human ears, all with normal hearing, underwent DPE testing. For each ear, DPE phase responses were computed for eight values of f2 varying from 10 to 0.78 kHz. Linear DPE phase versus DPE frequency relationships were found. Estimates of traveling wave delay from the ear canal to the f2 place varied from about 1 ms for the 10-kHz place to 3.5 ms for the 0.78 kHz place. These estimates agree well with previous traveling wave delay estimates using electrocochleography. Test-retest comparisons of delay estimates were generally within 0.25 ms. In addition, within-subject interaural delay differences were smaller than between-subject interaural differences. Within subject interaural delay differences were generally less than 0.5 ms. Male ears, when grouped together, had significantly longer delays (8%) to the 0.78-kHz place in comparison to female ears. The effect of DPE stimulus level on delay is presented for stimulus levels between 15 and 60 dB SPL. These data support the use of DPE phase responses as estimates of cochlear traveling wave delay. In comparison with electrophysiological and psychophysical techniques this method is purely cochlear-based and has the advantage of being rapid and noninvasive. PMID- 8408976 TI - Auditory nerve fiber representation of cues to voicing in syllable-final stop consonants. AB - Acoustic cues to the identity of consonants such as /d/ and /t/ vary according to contextual factors such as the position of the consonant within a syllable. However, investigations of the neural coding of consonants have almost always used stimuli in which the consonant occurs in the syllable-initial position. The present experiments examined the peripheral neural representation of spectral and temporal cues that can distinguish between stop consonants /d/ and /t/ in syllable-final position. Stimulus sets consisting of the syllables /hid/, /hit/, /hud/, and /hut/ were recorded by three different talkers. During the consonant closure interval, the spectrum of /d/ was characterized by the presence of a low frequency "voice bar." The closure interval for the voiceless consonant /t/ was longer and lacked a voice bar. Most neurons' responses were characterized by discharge rate decreases at the beginning of the closure interval and by rate increases that marked the release of the consonant closure. Exceptions were seen in the responses of neurons with characteristic frequencies (CFs) below approximately 0.7 kHz to syllables ending in /d/. These neurons responded to the voice bar with discharge rates that could approach the rates elicited by the vowel. The latencies of prominent discharge rate changes were measured for all neurons and used to compute the length of the "encoded closure interval." The encoded interval was clearly longer for syllables ending in /t/ than in /d/. The encoded interval increased with CF for both consonants but more rapidly for /t/. Differences in the encoded closure interval were small for syllables with different vowels or syllables produced by different talkers. PMID- 8408977 TI - Childhood behavior problems and bipolar disorder--relationship or coincidence? AB - A group of offspring at risk for Bipolar Disorder is compared to a Normal Control group whose parents had no psychiatric disorder and a group of offspring at risk for other, non-bipolar psychiatric disorder. Variables examined include childhood attention and behavior problems and psychopathology in young adulthood. Rates of childhood behavior and attention problems, and psychopathology and social/occupational impairment in young adulthood, were higher in the Bipolar Risk group than the Normal Control group, but no higher than in the non-bipolar (Combined Risk) group. Although childhood behavior and attention problems were significantly associated with other psychopathology in all three offspring groups, a unique relationship between childhood problems and young adult mood disorder was found only in the Bipolar Risk group. PMID- 8408978 TI - Affective comorbidity in panic disorder: is there a bipolar connection? AB - Although theoretical explanations for comorbidity in panic disorder (PD) abound in the literature, the complex clinical challenges of these patients have been neglected, especially where panic, obsessive-compulsive and 'soft' bipolar (e.g., hypomanic, cyclothymic and hyperthymic) conditions might co-exist. The aim of the present study has been to systematically explore the spectrum of intra-episodic and longitudinal comorbidity of 140 DSM-III-R PD patients--67.1% of whom concomitantly met the criteria for Agoraphobia--and who were consecutively admitted to the ambulatory service of the Psychiatric Clinic of the University of Pisa over a 2-year period. Comorbidity with strictly defined anxiety disorders- i.e., not explained as mere symptomatic extensions of PD--was relatively uncommon, and included Simple Phobia (10.7%), Social Phobia (6.4%), Generalized Anxiety Disorder (3.6%), and Obsessive-Compulsive Disorder (4.2%). Comorbidity with Major Depression--strictly limited to the melancholic subtype--occurred in 22.9%. Comorbidity with Bipolar Disorders included 2.1% with mania, 5% with hypomania, as well as 6.4% with cyclothymia, for a total of 13.5%; an additional 34.3% of PD patients met the criteria for hyperthymic temperament. We submit that such comorbid patterns are at the root of unwieldy clinical constructs like 'atypical depression' and 'borderline personality'. The relationship of panic disorder to other anxious-phobic and depressive states has been known for some time. Our data extend this relationship to soft bipolar disorders. Studies from other centers are needed to verify that the proposed new link is not merely due to referral bias to a tertiary university setting. PMID- 8408979 TI - Morning and evening light treatment of seasonal affective disorder: response, relapse and prediction. AB - Patients with seasonal affective disorder were randomly assigned to treatment with light in the morning (9.00-12.00 a.m.; n = 16; ML) or evening (6.00-9.00 p.m.; n = 11; EL). An intensive 24-day assessment procedure revealed the same response rates: 57% for ML, 50% for EL. During the rest of the winter season a relatively low relapse rate of 54% was found. No differences between ML and EL were found in the time course of depressed mood or fatigue. A significant negative correlation was found between diurnal variation during baseline and therapeutic response: the larger the diurnal variation the less the response, indicating a potential negative predictive value for this symptom. There were no significant correlations between baseline fatigue or hypersomnia and response. PMID- 8408980 TI - Monoamine oxidase inhibitors in resistant major depression. A double-blind comparison of brofaromine and tranylcypromine in patients resistant to tricyclic antidepressants. AB - In a double-blind study the selective monoamine oxidase-A inhibitor brofaromine was compared with the classical MAOI tranylcypromine in 39 patients with major depression resistant to treatment with tricyclic antidepressants. Concerning efficacy no significant differences were found. Ten out of 22 patients responded to brofaromine and 5 out of 17 patients to tranylcypromine. Adverse effects favoured brofaromine. Although orthostatic hypotension occurred in both groups, severe decrease in blood pressure and dizziness occurred significantly more with tranylcypromine. Both MAOIs caused a decrease in stage 4 and REM sleep and an increase in REM latency. In most patients receiving tranylcypromine REM sleep was completely abolished. PMID- 8408981 TI - Depression and T lymphocytes in patients with irritable bowel syndrome. AB - In 74 patients (23 male, 51 female) with irritable bowel syndrome (IBS) and in 15 matched control persons, an assessment of depression by means of the Zung Depression Self-Rating Scale (ZDS) and a determination of the number of total lymphocytes and T lymphocytes were carried out. In patients with IBS, the depression score was significantly higher than in controls. Among patients with IBS, the intensity of depressive symptoms did not relate to age, gender or to the type of the illness. IBS patients with a depression score on ZDS of 50 or more (31% of subjects) had a significantly lower number of total lymphocytes and T lymphocytes than the rest of IBS patients. The results may suggest a possible association between depression and indices of cellular immunity in IBS patients. PMID- 8408982 TI - The effect of anxiety induction on the regional uptake of 99mTc-exametazime in simple phobia as shown by single photon emission tomography (SPET). AB - Ten patients suffering from DSM-III-R simple phobia were studied under two conditions: (a) while listening to a 4 min relaxation tape, and (b) while listening to a 4 min audio tape describing exposure to the phobic stimulus. During each condition, subjects were injected with 99mTc-Exametazime, a marker of regional cerebral blood flow. Subjective and psychophysiological measures indicated a marked effect of the anxiety induction procedure. Ratio analysis of the SPET data revealed reductions in tracer uptake largely confined to posterior cerebral regions bilaterally. Analysis of brain regions of interest normalised to the whole brain slice showed reductions confined to right temporal/occipital regions. In general there was no clear association between subjective and physiological variables and changes in regional uptake of tracer as a consequence of the anxiety induction procedure. The changes in tracer uptake were dissimilar to those previously reported for other cognitive activation paradigms, providing some reassurance that those functional brain changes were not artefacts of non specific changes in state anxiety. These posterior brain changes may reflect alterations in activation of the GABA/benzodiazepine complex. PMID- 8408983 TI - Lack of effect of anxiety on total plasma MHPG in depressed patients. AB - This report was undertaken to test the noradrenergic deficiency hypothesis of depression and the postulated increase in noradrenergic activity associated to anxiety states. A possible dual effect of both depression and anxiety on total plasma MHPG levels was hypothesized and assessed in anxious and non-anxious depressed patients. The findings show a decrease in plasma MHPG levels in depressed patients whatever their degree of anxiety. There was no difference in total plasma MHPG levels either between anxious and non-anxious depressed patients or between low and high anxiety to depression ratio (ADR) depressed patients. Following antidepressant drug-treatment, a decrease in plasma MHPG was found. A positive correlation between the drug-induced decrease in NA activity and the severity of depression was observed, and suggested a relationship between the severity of depression and the instability of the NA system. No correlation between the drug-induced decrease in plasma MHPG and the degree of anxiety was found. The results do not suggest out an effect of anxiety on total plasma MHPG levels in depressed patients. PMID- 8408984 TI - Distinguishing instrumental and hostile aggression: does it make a difference? AB - An analogue task of instrumental and hostile aggression during a competitive game, modified to minimize overlap between aggressive responses, was evaluated in 8- to 14-year-old clinically referred boys (n = 33). Postgame interviews indicated that the hostile response, an aversive noise, was perceived by over 80% of subjects as hostile and not instrumental. In contrast, the instrumental response, blocking the opponent's game, was perceived about equally as having instrumental and hostile functions. The hostile aggressive response was uniquely correlated with continuous performance task impulsive commission errors (r = .51), which supported the theoretical relation of hostile aggression to poor impulse control. These results suggest that instrumental and hostile aggression can be distinguished and when precisely defined are distinct in theoretically important ways. PMID- 8408985 TI - British parents' beliefs about the causes of three forms of childhood psychological disturbance. AB - Parents were asked to rate which of 40 items were likely causes of one of three types of psychological disturbance common among 9-year-old children, the symptoms of which were outlined in vignettes. Parents' accounts were consistent and coherent. Symptoms of hyperactivity were linked to poor diet, antisocial conduct to a lack of discipline and unhappiness at school, and emotional problems to breakdowns in interpersonal relations and unhappiness at school. However, a number of factors (socioeconomic difficulties, attachment risks, and organic dysfunction) that research suggests are risks to development were rated by parents as unlikely causes of problems. The implications of this mismatch between "lay" and "expert" views is examined. PMID- 8408986 TI - Evidence for developmentally based diagnoses of oppositional defiant disorder and conduct disorder. AB - This paper compares the validity of DSM-III-R diagnoses of oppositional defiant disorder (ODD) and conduct disorder (CD) and an alternative option which is subdivided into three levels according to developmental sequence and severity: modified oppositional disorder (MODD), intermediate CD (ICD), and advanced CD (ACD). Using a sample of 177 boys followed over 3 years, both the DSM-III-R and the alternative diagnostic constructs are evaluated on three criteria: symptom discriminative validity, and diagnostic external and predictive validity. Most DSM-III-R ODD and CD symptoms discriminated between ODD and CD, but exceptions are noted. Additional analyses demonstrated considerable overlap among DSM-III-R oppositional symptoms. The majority of the symptoms proposed for the alternative option could be assigned to a specific level based on acceptable symptom discrimination. External validity lent support to the distinctions between DSM III-R ODD and CD, and between MODD, ICD, and ACD. MODD was a better predictor than ODD of which MODD, ICD, and ACD. MODD was a better predictor than ODD of which boys received a later diagnosis of CD. Suggestions are made for the inclusion and exclusion of symptoms for developmentally based diagnoses of oppositional and conduct disorders. PMID- 8408987 TI - Classical conditioning in children with attention deficit hyperactivity disorder (ADHD) and anxiety disorders: a test of Quay's model. AB - Quay (1988) put forward a model of childhood mental disorders based on Gray's (1982) theory that there exists within the brain a behavioral inhibition system (BIS), which processes signals related to aversive or punishing stimuli. According to this model, children with attention deficit hyperactivity disorder (ADHD) show lower than optimal levels of activity in this system, which leads to less responsiveness at a physiological level to signals related to punishment. Children with ADHD and controls were compared on a classical conditioning paradigm. Skin conductance and cardiac responses were measured in response to a conditioned stimulus that had been paired with an aversive unconditioned stimulus. There were no differences between the groups, suggesting that, in terms of classical conditioning, ADHD children are equally responsive to signals related to punishment as controls. PMID- 8408988 TI - The impact of synthesis teaching and parent training with mothers of conduct disordered children. AB - A procedure to enhance the impact of parent training with multistressed mother child dyads was explored with 29 clinic-referred dyads. The procedure, called synthesis teaching, was provided for one randomly selected group of dyads, while the other group was engaged in problem discussions about their children. Both groups received parent training as the principal intervention for the mothers' conduct-disordered children. Results showed that these interventions had no effects on dyads in the clinic setting, but had a progressively more significant effect at home for the synthesis teaching group. That is, mothers in this group showed reductions in their indiscriminate parenting and their children demonstrated behavioral improvements. In contrast, mothers in the control group did not change their behavior and their children did not demonstrate observed behavioral changes. Interpretations of the results center on how synthesis teaching might have produced these effects. PMID- 8408989 TI - The factor structure of ADHD items in DSM-III-R: internal consistency and external validation. AB - Previous research employing factor-analytic procedures to study the underlying dimensions of DSM-III attention deficit disorder with hyperactivity (ADDH) symptoms have consistently supported a two-factor model. Revision of the structure of the ADHD diagnosis in DSM-III-R, as well as inclusion of new items, has raised the question of comparability of the two diagnoses. To explore the significance of these changes, teacher ratings of DSM-III ADDH items and DSM-III R ADHD items of 85 nonreferred school children were factor-analyzed to determine their underlying factor structures. A similar two-factor solution was obtained for each diagnostic scale. The factors consisted of items believed to reflect inattention and hyperactivity-impulsivity constructs. These factors were further evaluated against results of a cognitive test battery to ascertain whether objective, external validation could be demonstrated. The hyperactivity impulsivity factor scores were related to continuous performance test measures of response inhibition, while inattention-disorganization factor scores were related to measures of attention and visual search. Implications for assessment and diagnosis of ADHD are discussed. PMID- 8408990 TI - Gender, creativity, depression, and attributional style in adolescents with high academic ability. AB - The present study examined the relationship among gender, creativity, depression, and attributional style among high-achieving adolescents. One hundred twenty eight eighth- and ninth-grade high-achieving students completed the Torrance Test of Creative Thinking (TTCT), the Children's Depression Inventory (CDI), and the Children's Attribution Style Questionnaire--Revised (KASTAN-R CASQ). The results indicate that there were gender differences only on the verbal component of the TTCT, with females scoring significantly higher. For both sexes, there was a significant relationship between figural creativity and a depressogenic attributional style. However, for females, high verbal creativity was associated with low levels of depression and a positive attributional style. PMID- 8408991 TI - Dental health care: today. PMID- 8408992 TI - Dental health care: tomorrow. PMID- 8408993 TI - Reflections on a career in dentistry. PMID- 8408994 TI - The development of a professional role orientation inventory. AB - This paper describes the development and testing of the Professional Role Orientation Inventory, a 40-item Likert-type instrument designed to assess a dentist's perception of his or her professional role. The test includes four 10 item scales representing four attitudinal dimensions: Authority, Responsibility, Agency, and Autonomy. The study describes differences between and among groups of professionals and dental students who took the test. Responses to the various scales suggest that practitioners do have different conceptualizations of their professional role, though these conceptions vary somewhat from the various models of professionalism described in the literature. PMID- 8408995 TI - A legacy of leadership. PMID- 8408996 TI - Study mission to India and Nepal. American People Ambassador Program. PMID- 8408997 TI - Dental health care: yesterday. PMID- 8408998 TI - Expanding the bubble. PMID- 8408999 TI - The rise of resin for cementing restorations. AB - Resin cements are popular and well-accepted for veneers, tooth-colored inlays, onlays and crowns, Maryland bridges and salvaging old useful crowns and fixed prostheses. Although used by a few practitioners for routine cementation of all crowns and fixed prostheses, it is doubtful that this trend will grow. A new category of cements--glass ionomer resin--soon will be available from several companies. It should be considered a routine cement as soon as it has proved itself. Its properties appear promising and nearly ideal. PMID- 8409000 TI - Altered passive eruption: the undiagnosed entity. AB - Excessive gingiva can result from delayed or altered tooth eruption. This condition should be considered in restorative dentistry, orthodontic and esthetic treatment. PMID- 8409001 TI - Estimated prevalence and distribution of reported orofacial pain in the United States. PMID- 8409002 TI - Managing patients with Alzheimer's. PMID- 8409003 TI - Reducing bacteria. PMID- 8409004 TI - Ridge augmentation and the elderly. PMID- 8409005 TI - What's new? PMID- 8409006 TI - Water vs. taxation. PMID- 8409007 TI - Post-traumatic TMD patients may require new treatment, say researchers. PMID- 8409008 TI - Occupational risks in dentistry: comforts and concerns. AB - Dentistry has been portrayed as a high-risk profession in recent reports. A look at several areas of concern, however, shows that the field is no more hazardous than any other profession--and less than most. PMID- 8409009 TI - HIV, health care workers and patients: how to ensure safety in the dental office. AB - What can be done to preserve the high standards of patient care, give reasonable assurances to patients and acknowledge the rights of health care workers? Conscientious adherence to infection control is the most responsible option available. PMID- 8409010 TI - Microbial contamination of dental unit waterlines: prevalence, intensity and microbiological characteristics. AB - Microbiological studies on dental unit water samples from 150 operatories revealed widespread and unacceptably high levels of contamination. Biofilm formation along the walls of the fine-bore waterlines is primarily responsible. These findings are reviewed and their relevance to contemporary infection control concerns discussed. PMID- 8409011 TI - HIV recovery from saliva before and after dental treatment: inhibitors may have critical role in viral inactivation. AB - Unstimulated whole saliva was collected from 21 HIV-positive men and women before and after dental treatment. The frequency of HIV detection did not increase after dental treatment. Infectious HIV was recovered from only one patient. Study findings raise the possibility that, in most cases, salivary inhibitors render the virus non-infectious. PMID- 8409012 TI - Effects of dental restorative materials: local and systemic responses reviewed. PMID- 8409013 TI - How personal protective equipment affects perceptions of dentists. AB - In this study, 248 college-educated young adults completed the Dental Fears Survey. Subjects then viewed one of two slides of a dentist. In one slide, the dentist wore no PPE at all; in the other, the dentist wore gloves, mask, glasses, face shield and lab coat. Subjects completed a 20-item semantic differential scale to assess their attitudes toward the dentist portrayed in the slide they saw. PMID- 8409014 TI - Alternative gingival retraction techniques and isolation of the cervical lesion. PMID- 8409015 TI - Much ado about nothing? PMID- 8409016 TI - Retention and the new patient myth. AB - The health of a dental practice cannot be determined by the number of new patients alone. There are several additional factors, including overhead, gross income and patient retention that also must be measured. Combining these factors and others, in addition to new patient numbers, is essential for gauging profitability. PMID- 8409017 TI - Formaldehyde in dentistry. PMID- 8409018 TI - Diagnosing caries. PMID- 8409019 TI - Receding gums. PMID- 8409020 TI - Sterilize it! PMID- 8409021 TI - Dr. Daniel Laskin receives Norton M. Ross Award. PMID- 8409022 TI - Studies show link between diabetes, severity of periodontal disease. PMID- 8409023 TI - Health system reform. What could it mean for dentistry? And what's ADA doing to get its point across? PMID- 8409024 TI - Dentistry and medicine. How they differ. And why that should make a difference in health system reform. PMID- 8409025 TI - Foreign reform. How selected other nations have changed their health systems for better or worse. PMID- 8409026 TI - Knowing the lingo. Health system reform has introduced a host of new terms to the lexicon. PMID- 8409027 TI - Self-disclosure of HIV infection to dentists and physicians. AB - We prospectively examined self-disclosure of HIV infection by 129 men and women to dentists and physicians. After an average of 2.3 years since initial HIV positive notification and repeated individual counseling, only 53 percent of subjects had told their dentists of HIV infection compared to 89 percent who had told their physicians. This high rate of non-disclosure despite extensive counseling raises concern about effectiveness of education alone in promoting voluntary self-disclosure. Our findings further support recommended universal precautions. PMID- 8409028 TI - Dentifrice use among preschool children. AB - Ingestion of fluoridated dentifrice by young children may be a major contributing factor to dental fluorosis, prevalent in the United States. Brushing habits of a small group of preschoolers were monitored to document parental involvement and amounts of dentifrice used. PMID- 8409029 TI - Glass ionomers: current clinical developments. PMID- 8409030 TI - Anticipating failure in extensive restorative treatment of the periodontally compromised patient. AB - If even one supporting tooth fails, a complex restorative treatment may have to be redone. Improved design of a restoration can anticipate failure and offer alterations to existing appliances. PMID- 8409031 TI - Enhancing infection control for elderly and medically compromised patients. AB - Some patients may be at risk for complications from relatively common infectious diseases. Influenza, tuberculosis and methicillin-resistant Staphylococcus aureus infection can lead to illness and even death in elderly, medically compromised and institutionalized individuals. Dental personnel caring for these individuals should adopt preventive strategies that are simple and inexpensive in addition to standard infection control guidelines. PMID- 8409032 TI - Posts, cores and patient care. AB - Custom cast posts and cores are quite acceptable, with proven usefulness. However, prefabricated posts and bonded composite or silver amalgam cores are replacing them rapidly. These posts and cores are faster and easier to construct than custom cast posts and cores, and they provide acceptable strength and serviceability at a relatively low cost to patients. PMID- 8409033 TI - The COMMIT dental model: tobacco control practices and attitudes. AB - The National Cancer Institute's program to help dentists reduce tobacco use among their patients is part of the Community Intervention Trial for Smoking Cessation, a 22-community, randomly controlled trial of an intervention program for smoking cessation. Results of COMMIT baseline surveys of dentists in the 11 intervention communities are presented. PMID- 8409034 TI - Embolic potential, prevention and management of mural thrombus complicating anterior myocardial infarction: a meta-analysis. AB - OBJECTIVES: The management of mural thrombus complicating acute anterior myocardial infarction remains controversial in part because of the small size of studies on this topic. We performed a meta-analysis of published studies to address three questions: 1) What is the embolic risk of mural thrombi after myocardial infarction? 2) What is the impact of systemic anticoagulation in reducing the embolic risk of mural thrombi? 3) What is the impact of systemic anticoagulation, thrombolytic therapy and antiplatelet therapy in preventing mural thrombus formation? METHODS: Studies were identified by a computerized and manual search and were included if they were published in manuscript form in the English-language literature. Pooling of data was performed by calculating the Mantel-Haenszel odds ratio and an event rate difference by the method of DerSimonian and Laird. RESULTS: The odds ratio for increased risk of emboli in the presence of echocardiographically demonstrated mural thrombus (11 studies, 856 patients) was 5.45 (95% confidence interval [CI] 3.02 to 9.83), and the event rate difference was 0.09 (95% CI 0.03 to 0.14). The odds ratio of anticoagulation versus no anticoagulation in preventing embolization (seven studies, 270 patients) was 0.14 (95% CI 0.04 to 0.52) with an event rate difference of -0.33 (95% CI -0.50 to -0.16). The odds ratio of anticoagulation versus control in preventing mural thrombus formation (four studies, 307 patients) was 0.32 (95% CI 0.20 to 0.52), and the event rate difference was -0.19 (95% CI -0.09 to -0.28). The odds ratio for thrombolytic therapy in preventing mural thrombus (six studies, 390 patients) was 0.48 (95% CI 0.29 to 0.79) with an event rate difference of -0.16 (95% CI 0.10 to -0.42), whereas for antiplatelet agents (two studies, 112 patients) the odds ratio was 1.43 (95% CI 0.04 to 56.8) with an event rate difference of 0.16 (95% CI -0.20 to 0.52). CONCLUSIONS: This analysis supports the hypotheses that 1) mural thrombus after myocardial infarction poses a significantly increased risk of embolization, 2) the risk of embolization is reduced by systemic anticoagulation, and 3) anticoagulation can prevent mural thrombus formation. Thrombolytic therapy may prevent mural thrombus formation, but evidence for a similar benefit of antiplatelet therapy is lacking. PMID- 8409035 TI - Extension of hypokinesia into angiographically perfused myocardium in patients with acute infarction. AB - OBJECTIVE: This study was performed to determine whether left ventricular hypokinesia due to acute myocardial infarction lies between the site of coronary artery occlusion and the end of the infarct-related artery in patients. BACKGROUND: Normalizing for the size of the risk region reduces variability in measuring infarct size in experimental studies. The ability to gauge the size of the region at risk of becoming dysfunctional may help reduce variability in measuring regional hypokinesia due to acute myocardial infarction. METHODS: Angiograms of 84 patients with acute infarction due to isolated stenosis of the right coronary artery (n = 40) or the left anterior descending coronary artery (n = 44) were analyzed. The location and length of the segment with hypokinesia more severe than -1 or -2 SD below the normal mean were determined by the centerline method. The risk region was defined as the left ventricular contour between the site of the occlusion and the end of the infarct-related artery on the angiogram. RESULTS: The segment with hypokinesia below -1 SD was longer than the risk region in 52% of patients with occlusion of the left anterior descending coronary artery, more frequently (p < 0.01) than in right coronary artery occlusion (22%), owing to extension of hypokinesia beyond the distal end of the artery. Extension of severe hypokinesia (below -2 SD) beyond the risk region occurred in 33% of patients with an anterior infarct and in 9% of patients with an inferior infarct. CONCLUSIONS: The size of the risk region cannot be assessed accurately from coronary angiography. PMID- 8409036 TI - Prolonged dilation with an autoperfusion balloon catheter for refractory acute occlusion related to percutaneous transluminal coronary angioplasty. AB - OBJECTIVES: Efficacy and safety of redilation by an autoperfusion balloon catheter over several hours were investigated in this retrospective and observational study. BACKGROUND: Acute occlusion, refractory to redilation, is a serious complication of coronary angioplasty. METHODS: Of 1,123 patients who underwent angioplasty, 83 had a refractory acute occlusion. Thiry-five patients were treated with extended dilation. Seven had stable, 19 unstable and 6 postinfarction angina and 3 had an acute infarction at the time of angioplasty. The duration of dilation was (mean +/- SD) 17 (+/- 6) h. RESULTS: Angiographically successful redilation, with a mean residual percent diameter stenosis of 13.5% (+/- 11.6%), was achieved in 22 (67.7%) of 34 patients. Five patients underwent bypass surgery. Three patients, who were poor surgical candidates, died. There was one new Q wave infarction and one death that occurred during extended dilation; one death and four operations were related to reocclusion immediately (< or = 30 min) after catheter withdrawal; and one death and one operation were related to in-hospital reocclusion. Overall success, defined as angiographic success and freedom from major events, was obtained in 20 (57%) of 35 patients (95% confidence interval 41% to 73%). Of the variables studied, only multilesion dilation was significantly (p = 0.018) associated with an unfavorable outcome. During a mean follow-up period of 13.8 (+/- 6.1) months, two patients underwent repeat angioplasty, one sustained an infarction and three underwent elective bypass surgery. CONCLUSIONS: In approximately half of the patients (20 [57%] of 35), an initial angioplasty failure due to refractory occlusion could be reverted to a successful procedure by prolonged dilation with an autoperfusion balloon catheter. PMID- 8409037 TI - Sequential intravascular ultrasound characterization of the mechanisms of rotational atherectomy and adjunct balloon angioplasty. AB - OBJECTIVES: The purpose of this study was to use sequential intravascular ultrasound imaging before intervention, after rotational atherectomy and after adjunct balloon angioplasty to characterize the mechanisms of lumen enlargement after each. BACKGROUND: Rotational atherectomy uses a high speed, rotating, diamond-tipped elliptic burr to abrade atherosclerotic plaque to increase lumen size. In vitro studies have shown that high speed rotational atherectomy selectively abrades hard, especially calcified, plaque elements. However, rotational atherectomy procedures usually require adjunct balloon angioplasty. METHODS: Forty-eight lesions in 46 patients were treated with rotational atherectomy followed by adjunct balloon angioplasty in 44. Quantitative coronary arteriographic and intravascular ultrasound measurements of the target lesion were made before intervention, after rotational atherectomy and after balloon angioplasty. RESULTS: Before intervention, target lesion external elastic membrane area measured 17.3 +/- 5.9 mm2, lumen area measured 1.8 +/- 0.9 mm2 and plaque plus media area measured 15.7 +/- 4.1 mm2. After rotational atherectomy, lumen area increased, plaque plus media area decreased, arc of target lesion calcium decreased and 26% of the target lesions had dissection planes. After adjunct balloon angioplasty, external elastic membrane area increased, lumen area increased, plaque plus media area did not change and 77% of the target lesions had dissection planes. Arterial expansion was seen in 80% of lesions. The pattern of dissection plane location, which was predominantly within calcified plaque after rotational atherectomy, became predominantly adjacent to calcified plaque after adjunct balloon angioplasty (p = 0.008). CONCLUSIONS: Sequential intravascular ultrasound imaging shows that high speed rotational atherectomy causes lumen enlargement by selective ablation of hard, especially calcific, atherosclerotic plaque with little tissue disruption and rare arterial expansion. Adjunct balloon angioplasty further increased lumen area by a combination of arterial dissection and arterial expansion, especially of compliant, noncalcified plaque elements. PMID- 8409038 TI - Myocardial risk area defined by technetium-99m sestamibi imaging during percutaneous transluminal coronary angioplasty: comparison with coronary angiography. AB - OBJECTIVES: The purpose of this study was to compare the assessment of myocardial area at risk in patients with coronary artery stenosis by coronary angiography and quantitative myocardial perfusion imaging with technetium-99m sestamibi. BACKGROUND: Decisions concerning patient management frequently rely on semiquantitative angiographic estimation of the myocardial area at risk, although this approach has not been well validated. Technetium-99m sestamibi is a perfusion imaging agent with little redistribution after initial myocardial uptake. This characteristic allows for injection during angioplasty and later imaging for visualization and quantitation of the nonperfused area at risk. METHODS: Thirty-nine patients referred for coronary angioplasty were studied. Technetium-99m sestamibi was injected intravenously during angioplasty balloon inflation. Planar (33 patients) or tomographic (6 patients) imaging was performed after completion of angioplasty. Imaging was repeated 24 to 48 h later. Myocardial risk area (perfusion defect on angioplasty image) was quantified as an integral using circumferential count distribution profiles and normal reference. Angiographic risk area was assessed using five scoring methods. RESULTS: The scintigraphic risk area was 14 +/- 15 on planar images and 39 +/- 16 on tomography. Scintigraphic risk area of patients with infarction was larger than in patients without (22 +/- 17 versus 7 +/- 8, p = 0.003). The left anterior descending coronary artery had a larger mean risk area than other vessels (22 +/- 15 versus 7 +/- 11, p = 0.002). The presence of angiographic collateral channels was associated with smaller risk areas. Angiographic risk scores correlated only moderately with the technetium-99m sestamibi risk area (r = 0.54 to 0.65), with considerable spread of data. CONCLUSIONS: Area at risk estimated from coronary angiography does not correlate well with that from quantitative myocardial perfusion imaging with technetium-99m sestamibi. These findings emphasize that the functional significance of coronary artery disease is not predicted by coronary anatomy alone. PMID- 8409039 TI - Radiation risk to patients from percutaneous transluminal coronary angioplasty. AB - OBJECTIVES: This retrospective study sought to estimate patient radiation exposure during percutaneous transluminal coronary angioplasty, the corresponding organ doses and the resulting cancer mortality risk. Patient demographic data were also examined. BACKGROUND: Coronary angioplasty is commonly used as an intervention for coronary atherosclerosis, and repeated application in the same patient is now common. The combined use of fluoroscopy and cineradiography in this complicated, delicate and, hence, lengthy procedure induced us to investigate the patient radiation exposures and resulting risks. METHODS: All complete records for angioplasty procedures performed over a 3-year period were entered into a data base. The data comprised 1,893 procedures performed in a total of 1,503 patients, of whom 21% had two or more procedures in the 3-year period. Fluoroscopy time was converted to entrance exposures, assuming a rate of 520 muC kg-1 min-1 (2.0 R min-1). Cineradiographic film lengths were determined for a smaller number of procedures (200) and converted to exposures at 7.7 muC kg 1 frame-1 (30 mR frame-1). In addition, fluoroscopy and cineradiographic times and, hence, exposures for 91 diagnostic angiograms performed in these patients were obtained. Exposures were converted to organ doses using the Monte Carlo results of the Rosenstein group and then to cancer mortality risks using the latest rates of the International Commission on Radiological Protection. RESULTS: The mean age was 56.0 years; men constituted 77.5% of the patients. Radiation doses varied considerably owing to a large spread in exposure times (e.g., fluoroscopy time per angioplasty case averaged 19 min but for some cases exceeded 1 h). The average patient skin entrance exposure per angioplasty procedure was 32.0 mC kg-1 (124 R), of which 69.7% was from cineradiography. The resulting cancer mortality risk per angioplasty procedure is approximately 8 x 10(-4). CONCLUSIONS: The skin exposures estimated for angioplasty are on average higher than for other X-ray procedures. The cancer mortality risk does not exceed the mortality risk of bypass surgery. Good professional practice requires maximization of the benefit/risk ratio through quality assurance in all aspects of the procedure. PMID- 8409040 TI - Economics of elective coronary revascularization. Comparison of costs and charges for conventional angioplasty, directional atherectomy, stenting and bypass surgery. AB - OBJECTIVES: This study was designed to evaluate more closely the true in-hospital costs of elective revascularization by directional coronary atherectomy and intracoronary stenting and to compare these costs with those of the traditional revascularization alternatives (i.e., conventional balloon angioplasty and coronary artery bypass surgery). BACKGROUND: Previous studies have suggested that total hospital charges for directional coronary atherectomy or intracoronary stenting are significantly higher than those for conventional angioplasty. However, hospital charges do not necessarily reflect true economic costs, and their use may provide misleading data with regard to cost-effectiveness. METHODS: We analyzed in-hospital charges from the itemized hospital accounts of 300 patients undergoing elective angioplasty, directional atherectomy, Palmaz-Schatz coronary stenting or bypass surgery between January 1, 1990 and December 31, 1991. Costs were then derived by adjusting itemized patient accounts for department-specific cost/charge ratios. Catheterization laboratory costs were based on actual resource consumption, and daily room costs were adjusted for the intensity of nursing services provided. RESULTS: Length of hospital stay was similar for atherectomy (2.3 +/- 1.5 days) and conventional angioplasty (2.6 +/- 1.7 days) but significantly longer for stenting (5.5 +/- 2.6 days, p < 0.05). Total costs were also significantly higher for coronary stenting ($7,878 +/- $3,270, median $6,699, p < 0.05) than for angioplasty ($5,396 +/- $2,829, median $4,753) or atherectomy ($5,726 +/- $2,716, median $4,986). However, length of stay, resource consumption (laboratory and radiologic testing, drugs, blood products, for example) and total costs for bypass surgery were still greater than for any of the percutaneous interventional procedures. CONCLUSIONS: In contrast to previous studies utilizing only hospital charges, the in-hospital costs of angioplasty and directional coronary atherectomy were similar. Although the cost of coronary stenting was approximately $2,500 higher than that of conventional angioplasty, the magnitude of this difference was smaller than the $6,300 increment previously suggested on the basis of analysis of hospital charges. These findings reflect the inherent discrepancies between cost-based and charge based methodologies and may have important implications for future studies evaluating the relative cost-effectiveness of newer coronary interventions. PMID- 8409041 TI - Argentine randomized trial of percutaneous transluminal coronary angioplasty versus coronary artery bypass surgery in multivessel disease (ERACI): in-hospital results and 1-year follow-up. ERACI Group. AB - OBJECTIVES: This study was designed to compare freedom from combined cardiac events (death, angina, myocardial infarction) at 1-, 3- and 5-year follow-up in patients with multivessel disease randomized to either percutaneous transluminal coronary angioplasty or coronary artery bypass graft surgery. BACKGROUND: Percutaneous transluminal coronary angioplasty has been an effective approach in patients with coronary artery disease, but its role in patients with multivessel coronary artery disease is still controversial. METHODS: One-hundred twenty-seven patients with multivessel disease and lesions suitable for either form of therapy were randomized to either coronary artery bypass grafting (n = 64) or coronary angioplasty (n = 63). In this study we report the immediate results and freedom from combined cardiac events at 1-year follow-up. RESULTS: Demographic, clinical and angiographic characteristics were similar in both groups. There were no differences in in-hospital deaths, frequency of periprocedure myocardial infarction or need for emergency revascularization procedures between the two groups. At 1-year follow-up, there were no differences in mortality or in the incidence of myocardial infarction between the groups. However, patients treated with coronary artery bypass grafting were more frequently free of angina, reinterventions and combined cardiac events than were patients treated with coronary angioplasty (83.5% vs. 63.7%, p < 0.005). In-hospital cost and cumulative cost at 1-year follow-up were greater for the coronary artery bypass grafting than for the coronary angioplasty group. CONCLUSIONS: No significant differences were found in major in-hospital complications between patients treated with coronary artery bypass grafting or coronary angioplasty. Although at 1-year follow-up there were no differences in survival and freedom from myocardial infarction, patients in the coronary artery bypass grafting group were more frequently free from angina, reinterventions and combined events than were patients in the coronary angioplasty group. PMID- 8409042 TI - Variability in measures of coronary lumen dimensions using quantitative coronary angiography. AB - OBJECTIVES: The purpose of this study was to determine the true total variability of quantitative coronary angiographic measures and their components in the clinical setting. BACKGROUND: Many studies describe quantitative coronary angiographic variability on the basis of repeated quantitative coronary angiographic measures from the same cineangiogram. Although these studies characterize well the performance of quantitative coronary angiographic analysis methods, they do not include other potentially important sources of variability in results of this procedure, such as day to day variations in patients and equipment or variability in selection of frames for analysis. METHODS: Coronary angiograms from 20 patients who underwent diagnostic angiography followed by percutaneous transluminal coronary angioplasty an average of 2.9 days later were reviewed. A total of 30 lesions well visualized in both films were analyzed multiple times using an automated first-derivative edge-detection quantitative coronary angiographic technique. RESULTS: The coefficient of variation for quantitative coronary angiographic measures of the same lesions from separate angiograms ranged from 8.11% to 14.01%. Average diameter was the least variable and percent diameter stenosis the most variable. Day to day variations in the patient, procedure and equipment accounted for an average of 30% of the total variability. Of the remaining variability, only 13.26% was due to variability in frame selection. CONCLUSIONS: These results provide useful information for planning clinical studies using quantitative coronary angiography, identify areas where additional improvements in this technology are needed and define more clearly the applicability of quantitative coronary angiography in the setting of routine clinical practice. PMID- 8409043 TI - Safety of thrombolytic therapy in elderly patients with massive pulmonary embolism: a comparison with nonelderly patients. AB - OBJECTIVES: The aim of the study was to prospectively estimate the safety of thrombolytic therapy in elderly patients with massive pulmonary embolism in comparison with that in nonelderly patients. BACKGROUND: In massive pulmonary embolism, lysis of thrombi can be achieved faster with thrombolytic therapy than with conventional heparin therapy, but it is administered with great caution in elderly patients because the risk of bleeding is thought to be higher than in nonelderly patients. Yet, thrombolytic therapy might be of value in elderly patients also, in allowing potentially more rapid improvement than is achieved with conventional heparin therapy. METHODS: Eighty-nine patients with massive pulmonary embolism defined as Miller score > or = 17/34 underwent thrombolytic therapy without consideration of age if they had no contraindication for such treatment. Fifty-three patients were < or = 70 years old (mean age +/- SD 54 +/- 15 years; range 18 to 70), and 36 patients were > or = 71 years old (78 +/- 5 years; range 71 to 88). Except for mean age, there were no significant differences between the two treatment groups, particularly in terms of clinical presentation, average Miller score and pulmonary artery pressure regimen. Thrombolytic therapy was administered in the form of streptokinase at a dose of 100,000 IU/h over 12 h, with an initial injection of 250,000 IU over 15 min. Heparin was introduced 12 h after initiation of thrombolytic therapy. Urokinase or tissue-type plasminogen activator was used only in case of contraindication to streptokinase. RESULTS: The frequency of uncomplicated clinical course was the same in both treatment groups. Surgical embolectomy was necessary in three nonelderly patients (5.6%) and one elderly patient (2.7%). Changes in pulmonary pressure regimen and Miller score were identical in both groups. Three patients died during the in-hospital course: two nonelderly patients (3.7%) and one elderly patient (2.7%). Minor bleeding occurred in five nonelderly (9.4%) and five elderly (13.8%) patients (p = 0.74). Major bleeding was observed in three nonelderly (5.6%) and five elderly (13.8%) patients (p = 0.29). Bleeding subsequent to early invasive procedure accounted for six (75%) of eight patients with major bleeding: two nonelderly patients (one of whom died) and four elderly patients. No intracranial hemorrhage was observed. No predisposing factor for bleeding was identified, except the need for early vascular access for pulmonary angiography through the femoral approach or for percutaneous insertion of an intracaval device for partial interruption of the inferior vena cava. CONCLUSIONS: Thrombolytic therapy administered for massive pulmonary embolism in patients free of contraindication yields similar results and carries a similar risk for bleeding complications in elderly compared with nonelderly patients. Limiting early invasive procedures may result in less frequent major bleeding complications. PMID- 8409044 TI - Biologic effects of recombinant hirudin (CGP 39393) in human volunteers. European Hirudin in Thrombosis Group. AB - OBJECTIVES: The purpose of this study was to investigate the biologic efficacy and pharmacokinetics of different doses of recombinant hirudin administered in single or repeated subcutaneous injections in healthy volunteers. BACKGROUND: Hirudin is a highly specific inhibitor of thrombin, the pivotal enzyme in thrombosis. Differences between hirudin and heparin in experimental animals indicate that hirudin may be a superior antithrombotic drug in humans. METHODS: The biologic effect of recombinant desulfato-hirudin (CGP 39393) administered as single or repeated (every 8 h for 3 days or every 12 h for 6 days) subcutaneous injections was studied in 231 healthy human volunteers. RESULTS: Single subcutaneous doses of 0.1, 0.2, 0.3, 0.4, 0.5 and 0.75 mg/kg body weight in 195, 8, 12, 8, 4 and 4 volunteers, respectively, prolonged the activated partial thromboplastin time in a dose-proportional fashion within the 1st 30 min, with a near-maximal effect for 3 to 4 h after the dose. The mean activated partial thromboplastin time increased to 1.48 and 1.93 times baseline values 30 min after single subcutaneous injections of 0.2 and 0.4 mg/kg of CGP 39393, respectively. There was a linear relation over a wide range between the activated partial thromboplastin time prolongation and plasma concentrations of CGP 39393. Plasma clearance was between 1.5 and 1.7 ml/min per kg. The subcutaneous administration of 0.3 and 0.5 mg recombinant hirudin three times a day for 3 days or two times a day for 6 days prolonged the activated partial thromboplastin time by 1.71 to 1.69 and 1.78 to 1.92 times baseline levels, respectively, with the preinjection values maintained in the hypocoagulable range. No prolongation of bleeding time was measured at peak plasma hirudin levels. Because thrombin and prothrombin times are not able to reflect high or low CGP 39393 concentrations, respectively, neither test is suitable for monitoring administration of this drug. CONCLUSIONS: CGP 39393 appears to be well tolerated in volunteers, even after repeated doses. The activated partial thromboplastin time test seems to be well suited to monitor the anticoagulant effect of recombinant hirudin because the dose effect is linear up to 0.5 mg/kg of subcutaneous CGP 39393. The prolongation of activated partial thromboplastin time after subcutaneous injection of CGP 39393 shows a plateau lasting for 3 h. Further studies are now required to determine the dose that will provide the best antithrombotic effect and the lowest bleeding tendency in arterial or venous thrombosis indications. PMID- 8409045 TI - Selective thrombin inhibitors: the next generation of anticoagulants. PMID- 8409046 TI - Use of a regional wall motion score to enhance risk stratification of patients receiving an implantable cardioverter-defibrillator. AB - OBJECTIVES: We postulated that preoperative assessment of both regional wall motion and left ventricular ejection fraction would serve as an accurate prognostic indicator of long-term cardiac mortality and functional outcome in patients treated with an implantable cardioverter-defibrillator. BACKGROUND: Long term cardiac mortality has remained high in patients receiving an implantable cardioverter-defibrillator. The ability to risk stratify patients before defibrillator implantation is becoming increasingly important from a medical and economic standpoint. METHODS: The hypothesis was retrospectively tested in 74 patients who had received an implantable cardioverter-defibrillator. Left ventricular ejection fraction and regional wall motion score, derived from centerline chord motion analysis, were calculated for each patient from the preoperative right anterior oblique contrast ventriculogram. Wall motion score was the only significant independent predictor of long-term cardiac mortality and functional status by multivariate analysis because of its enhanced prognostic capability in patients with an ejection fraction in the critical range of 30% to 40%. RESULTS: Patients with an ejection fraction > 40% had a 3-year cardiac mortality rate of 0% compared with 25% for those with an ejection fraction of 30% to 40% and 48% for those with an ejection fraction < 30% (p < 0.05). Similarly, 75% of patients with an ejection fraction > 40% were in New York Heart Association functional class I or II during long-term follow-up compared with 59% of those with an ejection fraction 30% to 40% and 29% of those with an ejection fraction < 30%. Among patients with an ejection fraction of 30% to 40%, those with a wall motion score > 16% had a 3-year cardiac mortality rate of 0% compared with 71% of those with a wall motion score < or = 16% (p = 0.002). In addition, 86% of patients with a wall motion score > 16% were in functional class I or II during long-term follow-up compared with 13% of those with a wall motion score < or = 16% (p = 0.001). CONCLUSIONS: Long-term cardiac mortality and functional outcome in patients receiving an implantable cardioverter-defibrillator can be predicted if the left ventricular ejection fraction and regional wall motion score are measured preoperatively. PMID- 8409047 TI - Recognition and catheter ablation of subepicardial accessory pathways. AB - OBJECTIVES: The purpose of this study was to characterize left-sided accessory pathways that traverse the atrioventricular (AV) groove subepicardially and to describe results of radiofrequency catheter ablation within the coronary sinus in the patients studied. BACKGROUND: Radiofrequency catheter ablation has proved to be a safe and effective method for treatment of accessory pathways; however, subepicardial accessory pathways may account for some of the failures encountered during endocardial ablation. METHODS: The study group comprised 51 consecutive patients with a left-sided accessory pathway who were undergoing radio-frequency catheter ablation. Initially, the ablation catheter was introduced into a femoral artery and positioned on the ventricular aspect of the mitral annulus. If this endocardial approach was unsuccessful, the ablation catheter was introduced into the coronary sinus and energy applied at sites with shorter activation times than those recorded from the endocardium. RESULTS: Five (10%) of 51 patients with a left-sided accessory pathway could not have accessory pathway conduction interrupted with a median of 18 endocardial radiofrequency energy applications. Accessory pathway potentials were less frequent during endocardial mapping in these 5 patients than in the 46 patients whose accessory pathway was successfully ablated from the endocardial surface. All five of these patients later had successful ablation using one or two applications of radiofrequency energy from within the coronary sinus. Effective target site electrograms in the coronary sinus were characterized by an accessory pathway potential that was larger than the corresponding atrial or ventricular electrogram. There were no complications or recurrences after ablation within the coronary sinus. CONCLUSIONS: Some left sided accessory pathways may be difficult to ablate from the endocardial surface because they traverse the AV groove subepicardially. The absence of an accessory pathway potential during endocardial mapping in combination with a relatively large accessory pathway potential within the coronary sinus may be a useful marker of a subepicardial pathway. In this select group of patients, radiofrequency catheter ablation from within the coronary sinus appears to enhance efficacy. PMID- 8409048 TI - Multipolar endocardial mapping of the right atrium during cardiac catheterization: description of a new technique. AB - OBJECTIVES: Using a new mapping system that allows the simultaneous acquisition of data from 25 right atrial bipolar electrodes during cardiac catheterization, we mapped normal sinus rhythm and atrial reentrant tachycardia in 24 sheep (20 to 49 kg) and 7 pigs (25 to 35 kg). BACKGROUND: Rapid, high resolution mapping during cardiac catheterization may shorten ablation procedures and permit ablation of otherwise refractory arrhythmias. METHODS: A flexible, elliptic, basket-shaped recording catheter has five spokes, each with 10 electrodes arranged as 5 bipolar pairs. Catheter shape, electrode spacing and introduction technique were modified in response to the results of experiments in the first 23 animals. In the most recent eight animals, retraction of a string attached to the distal tip distended the basket, providing safe tissue contact. Filtered (30 to 250 Hz) bipolar recordings from all 25 electrode pairs, as well as a surface electrocardiogram, were recorded and digitized at 1,000 Hz using custom software. An activation map was digitally constructed and superimposed on anteroposterior and lateral fluoroscopic catheter images. Bipolar recordings were made in normal sinus rhythm (31 animals), with adequate signals recorded from > 95% of electrode pairs. Rapid burst pacing and intentional right atrial air embolus (30 to 50 ml) induced sustained atrial reentrant tachycardia in five animals, which was also adequately recorded. RESULTS: Catheter positioning and complete atrial mapping required < 10 min after venous access in the most recent eight experiments. The catheter was left in position for up to 4 h. Postmortem evaluation revealed minor superficial abrasion of the venae cavae or right atrial endocardium in six animals and moderate abrasion in two. No other damage was observed. CONCLUSIONS: This new system may ultimately assist in mapping simple or complex atrial arrhythmias during cardiac catheterization. PMID- 8409049 TI - Asymptomatic ventricular arrhythmias and mortality risk in subjects with left ventricular hypertrophy. AB - OBJECTIVES: The purpose of this study was to evaluate the long-term prognostic role of asymptomatic ventricular arrhythmias in original Framingham Heart Study subjects and Framing-ham Offspring Study subjects who had echocardiographic evidence of left ventricular hypertrophy. BACKGROUND: Echocardiographically determined left ventricular hypertrophy is associated with increased risk for ventricular arrhythmias. There are no population-based data available with regard to the long-term prognostic implications of asymptomatic ventricular arrhythmias in subjects with left ventricular hypertrophy. METHODS: In a population-based cohort study, we studied 224 men and 393 women with echocardiographically determined left ventricular hypertrophy who were free of coronary heart disease and had 1-h ambulatory electrocardiograms at the baseline examination. The age adjusted prevalence of complex or frequent ventricular arrhythmias (> 30 ventricular premature beats/h, multiform premature complexes, couplets, ventricular tachycardia or R on T ventricular premature complexes) was 28% (51 of 224) in men and 17% (71 of 393) in women. The mean follow-up period was 5.7 years for cohort and 4.5 years for offspring subjects. RESULTS: In men with complex or frequent arrhythmias, the 6-year cumulative incidence of all-cause mortality was 38%, whereas in those free of arrhythmia it was 12%; corresponding values in women were 22% and 11%. The cumulative incidence of myocardial infarction or death due to coronary heart disease was 20% for men with and 10% for men without arrhythmia, but in women little difference was noted (5% vs. 4%). After adjustment for age and gender in a Cox proportional hazards model, subjects with complex or frequent arrhythmia were at increased risk for all-cause mortality (hazard ratio 1.80, 95% confidence interval [CI] 1.13 to 2.87, p = 0.013). After adjusting for eight clinical covariates, the increased risk for all-cause mortality remained marginally significant (hazard ratio 1.62, 95% CI 0.98 to 2.68, p = 0.058). No significant increased risk was noted for myocardial infarction or death due to coronary heart disease. CONCLUSIONS: In subjects with left ventricular hypertrophy, the presence of asymptomatic ventricular arrhythmias was associated with higher mortality, which was statistically significant after adjusting for age and gender and marginally significant after taking into account other covariates. PMID- 8409050 TI - Multiple monomorphic ventricular tachycardia configurations predict failure of antiarrhythmic drug therapy guided by electrophysiologic study. AB - OBJECTIVES: The purpose of this study was to determine whether the induction at electrophysiologic study of sustained monomorphic ventricular tachycardias with multiple QRS complex configurations predicted failure of subsequent serial electrophysiologic study guided antiarrhythmic drug testing. BACKGROUND: Ventricular tachycardias with multiple QRS complex configurations are associated with failure of surgical therapy for ventricular tachycardia. As such, the presence of multiple monomorphic QRS complex ventricular tachycardias during electrophysiologic testing may predict failure of subsequent medical therapy. METHODS: Fifty-one consecutive patients with coronary artery disease had reproducible induction of monomorphic ventricular tachycardia during a baseline electrophysiologic study. Each patient then underwent a mean of 1.5 antiarrhythmic drug trials. An antiarrhythmic drug regimen that suppressed induction of ventricular tachycardia was identified in 13 (26%) of the 51 patients. RESULTS: Patients with only one inducible monomorphic QRS complex ventricular tachycardia at baseline study were more likely to have an antiarrhythmic drug regimen identified that suppressed inducible ventricular tachycardia than were patients with multiple monomorphic QRS complex ventricular tachycardias (12[36%] of 33 patients vs. 1 [6%] of 18, p = 0.04). In seven patients with only one induced configuration of ventricular tachycardia, a second monomorphic ventricular tachycardia with a different QRS complex configuration occurred during attempts at pacing termination of the induced ventricular tachycardia. None of these seven patients then had successful drug suppression of inducible ventricular tachycardia. Thus, 12 (46%) of 26 patients with a single monomorphic QRS complex ventricular tachycardia observed at baseline study had successful serial drug testing compared with 1 (4%) of 25 patients with multiple QRS complex ventricular tachycardia configurations (p = 0.002). CONCLUSIONS: The induction or observation of multiple monomorphic QRS complex ventricular tachycardias at baseline electrophysiologic study predicted failure of subsequent serial electrophysiologic study--guided antiarrhythmic drug therapy. PMID- 8409051 TI - Role of autonomic reflexes in syncope associated with paroxysmal atrial fibrillation. AB - OBJECTIVES: The purpose of this study was to evaluate the role of autonomic reflexes in the genesis of syncope associated with the onset of paroxysmal atrial fibrillation. BACKGROUND: Syncope associated with paroxysmal atrial fibrillation has been interpreted as an ominous finding predictive of rapid ventricular rates. However, various mechanisms may be involved when heart rate is not particularly high. METHODS: Forty patients (age 60 +/- 14 years, 20 men, 20 women) with syncope and atrial fibrillation were compared with atrial fibrillation without syncope. Carotid sinus massage and head-up tilt testing (at 60 degrees for 60 min at baseline and during isoproterenol infusion) were performed during sinus rhythm. A positive response was defined as the induction of syncope. Atrial fibrillation was also induced on a tilt table at 60 degrees by means of short bursts of atrial pacing. RESULTS: Results of carotid sinus massage were positive in 15 (37%) of 40 patients but in no control subjects (p = 0.002). Head-up tilt test findings were positive in 25 (66%) of 38 patients and in 2 (12%) of 16 control subjects (p = 0.0004). The induction of atrial fibrillation in the upright position elicited syncope in 16 (42%) of 38 patients but in none of 16 control subjects (p = 0.001). At the beginning of atrial fibrillation, systolic blood pressure was lower in patients than in control subjects (88 +/- 32 vs. 127 +/- 32 mm Hg), whereas mean heart rate was similar (142 +/- 35 vs. 134 +/- 25 beats/min). The correlation between heart rate and systolic blood pressure was weak (r = 0.35), and in five patients syncope occurred at a heart rate < or = 130 beats/min. At the time of syncope, heart rate decreased (-12 +/- 21 beats/min) in patients with induced syncope, whereas it remained unchanged in patients without induced syncope (+1 +/- 17 beats/min, p = 0.04) or slightly increased in control subjects (+9 +/- 21 beats/min, p = 0.009). CONCLUSIONS: Patients with syncope associated with paroxysmal atrial fibrillation are predisposed to an abnormal neural response during both sinus rhythm and arrhythmia. In some patients the onset of atrial fibrillation triggers vasovagal syncope. PMID- 8409052 TI - An abnormal neural reflex plays a role in causing syncope in sinus bradycardia. AB - OBJECTIVES: This study investigates the role of an abnormal neural reflex in causing syncope in patients with sinus bradycardia. BACKGROUND: Syncope is commonly considered an indication of severity in sinus bradycardia. However, the occurrence of syncope is unpredictable, and the prognosis appears to be similar in patients with and without syncope. METHODS: Head-up tilt testing (60 degrees for 60 min), carotid sinus massage in the supine and standing positions, 24-h Holter ambulatory electrocardiographic (ECG) recording and electrophysiologic study before and after pharmacologic autonomic blockade were performed in 25 patients with sinus bradycardia and syncope (group I, sinus rate < 50 beats/min, age 71 +/- 12 years) and 25 patients with sinus bradycardia and no neurologic symptoms (group II, sinus rate < 50 beats/min, age 67 +/- 16 years). RESULTS: Clinical characteristics and ambulatory ECG monitoring data were similar in the two study groups. A positive response (induction of syncope or presyncope with hypotension and/or bradycardia) was obtained by head-up tilt testing in 15 group I (60%) and in 3 group II (12%) patients (p < 0.001) and by carotid sinus massage in 11 group I (44%) and 6 group II (24%) patients (p = NS). Results of at least one test (head-up tilt testing or carotid sinus massage, or both) were positive in 19 group I (76%) and 9 group II (36%) patients (p < 0.01). Basal and intrinsic corrected sinus node recovery time did not differ significantly between the two groups. An abnormal intrinsic heart rate was present in 66% of group I and 26% of group II patients (p < 0.01). The different percentage of positive findings on head-up tilt testing and carotid sinus massage in the two groups was independent of the presence of intrinsic sinus node dysfunction. CONCLUSIONS: These results indicate that an abnormal neural reflex plays a role in causing syncope in patients with sinus bradycardia. This reflex seems to be unrelated to the severity of sinus node dysfunction, even if the latter could enhance the cardioinhibitory response. PMID- 8409053 TI - Unique sensing errors in third-generation implantable cardioverter defibrillators. AB - OBJECTIVES: Third-generation cardioverter-defibrillators appear to be susceptible to unique sensing errors. This study was performed to determine the incidence and types of sensing errors in combination therapy implantable devices. BACKGROUND: One of the advantages offered by third-generation implantable cardioverter defibrillators is the combination of bradycardia and antitachycardia pacing and cardioversion-defibrillation capabilities in a single device. The potential for unique sensing errors, those caused by the conflicts presented by combining bradycardia and tachycardia sensing and therapy algorithms in the same device, has not been previously addressed. METHODS: To determine the incidence of important sensing errors, 61 patients with a combination therapy device (Cadence [Ventritex] and PCD [Medtronic]) were studied for a 25-month period. In addition to surface electrocardiographic recordings during implantation and routine device testing, real-time and stored electrograms recorded from the rate-sensing leads (Cadence) and real-time marker channel recordings (PCD) were reviewed to diagnose sensing errors that resulted in symptoms, device inefficacy or delivery of inappropriate therapy. After recognition, specific reprogramming steps were performed in an attempt to avoid recurrent sensing errors. RESULTS: A total of 13 sensing errors were diagnosed in 12 patients (19.7%); the incidence was similar in both devices. Five distinct categories of sensing errors were identified. After device reprogramming, only one recurrent error occurred in 98 patient months of follow-up. CONCLUSIONS: Important sensing errors occur in approximately 20% of patients with third-generation combination therapy cardioverter defibrillators. Prompt diagnosis of sensing errors can lead to specific reprogramming steps to avoid recurrent errors. PMID- 8409054 TI - Five-year angiographic follow-up of factors associated with progression of coronary artery disease in the Coronary Artery Surgery Study (CASS). CASS Participating Investigators and Staff. AB - OBJECTIVES: The Coronary Artery Surgery Study (CASS) required participants to undergo follow-up angiography at 5 years to identify clinical and angiographic features associated with progression of coronary artery disease. BACKGROUND: The CASS randomized 780 patients at 11 participating clinical centers between an initial strategy of medical therapy versus bypass surgery. Five clinical sites accomplished follow-up angiography in > 50% of their randomized subjects within a 42- to 66-month period after the entry arteriogram (n = 314). METHODS: Qualified clinical site angiographers, using side by side film review, evaluated an average of 13 segments/patient on both arteriograms for initial stenosis severity, morphologic features, lesion location and occurrence of disease progression or occlusion. Progression was defined as further definite narrowing by > or = 15% and occlusion as lesion progression to > or = 98%. Lesions were subcategorized as to whether they were univariate and had or had not been treated with bypass surgery. Multivariate logistic regression analyses were performed. RESULTS: For nonbypassed segments, right coronary artery and left anterior descending artery proximal and midlocations were associated with disease progression. For stenosis containing segments, the initial severity, a non-left anterior descending artery location and increased treadmill duration predicted progression. Segment occlusion was associated with initial lesion severity, right coronary artery location and subsequent interval myocardial infarction. There were few predictors of progression or occlusion in bypassed arteries, other than initial lesion severity. CONCLUSIONS: Univariate and multivariate associations with lesion progression and occlusion included diabetes, lesion location, elevated cholesterol level, interval infarction and lesion morphology. These angiographic results, collected in a prospective trial, are consistent with known risk factors. PMID- 8409055 TI - Smoking cessation therapy for the patient with heart disease. PMID- 8409056 TI - Echocardiographic characterization of the improvement in right ventricular function in patients with severe pulmonary hypertension after single-lung transplantation. AB - OBJECTIVES: This study was designed to characterize immediate, early and long term changes in right ventricular structure and function, as defined by two dimensional and Doppler echocardiography, after single-lung transplantation in patients with severe pulmonary hypertension. BACKGROUND: Single-lung transplantation has recently been shown to dramatically improve hemodynamics in patients with primary pulmonary hypertension who had unsuccessful medical therapy. METHODS: Fourteen patients with severe pulmonary hypertension who underwent single-lung transplantation were studied with transthoracic and transesophageal two-dimensional and Doppler echocardiography. Right ventricular dimensions were measured in the apical four-chamber view. Right ventricular ejection and acceleration times and peak velocity of tricuspid regurgitation were measured by Doppler study. Results of right heart catheterization were available early (< 3 months) after transplantation in 10 of 13 patients and late after transplantation (6 months to 2 years) in 11 patients. RESULTS: In the early posttransplantation studies, right ventricular dimensions decreased and fractional area change and ejection fraction increased in all patients, but right ventricular wall thickness did not change significantly. Tricuspid regurgitation lessened markedly in all patients. Long-term decreases in right ventricular dimension and improvement in systolic function were sustained. Right ventricular wall thickness significantly decreased compared with the early postoperative value (0.76 +/- 0.1 cm compared with 0.63 +/- 0.14 cm, p < 0.02). CONCLUSIONS: Two-dimensional echocardiography demonstrates sustained improvement in right ventricular function after single-lung transplantation for severe pulmonary hypertension despite severe preoperative dysfunction. PMID- 8409057 TI - New subtype of apical hypertrophic cardiomyopathy identified with nuclear magnetic resonance imaging as an underlying cause of markedly inverted T waves. AB - OBJECTIVES: The aim of this study was to elucidate the clinical importance of a new subtype of apical hypertrophic cardiomyopathy that could not be diagnosed with the classical diagnostic criteria. BACKGROUND: Apical hypertrophic cardiomyopathy is recognized by a characteristic spade-shaped intraventricular cavity on the end-diastolic left ventriculogram in the right anterior oblique projection, often associated with giant negative T waves [negativity > or = 1.0 mV (10 mm)]. As an underlying cause of giant negative T waves, an additional new subtype of apical hypertrophic cardiomyopathy has been identified. METHODS: In 40 patients with inverted T waves (negativity > or = 0.5 mV), including 26 patients with giant negative T waves, nuclear magnetic resonance (NMR) long-axis images corresponding to the left ventriculogram in the right anterior oblique projection and short-axis images at various levels, including the apical level, were obtained to define the site of hypertrophied myocardium. RESULTS: Long-axis images indicated a spadelike configuration in 17 patients, whereas this diagnostic configuration was not present in the other 23 patients. Nine of these 23 patients had significantly hypertrophied myocardium at the basal level. In the 14 remaining patients, short-axis images indicated no hypertrophy at the basal level and proved that the area of hypertrophied myocardium was confined to a narrow region of the septum or the anterior or lateral wall at the apical level (nonspade apical hypertrophic cardiomyopathy). The hypertrophied myocardium of the nonspade type was so narrowly confined that the mass did not form a spadelike configuration or could not be detected on the long-axis image. CONCLUSIONS: Nonspade apical hypertrophic cardiomyopathy was newly identified on NMR short axis images, and this could be an additional, important underlying cause of moderately to severely inverted T waves. PMID- 8409058 TI - Regional ventricular wall motion abnormalities in tricuspid atresia after the Fontan procedure. AB - OBJECTIVES: The purpose of this study was to determine whether wall motion abnormalities are present before or after the Fontan procedure in patients with a univentricular heart of the left ventricular type with an absent right atrioventricular valve connection (tricuspid atresia) and to assess the impact of such abnormalities on ventricular performance and clinical outcome. BACKGROUND: Normal systolic and diastolic ventricular function is critical for a successful Fontan repair. However, there have been no previous studies addressing the relation between regional ventricular function and hemodynamic factors. METHODS: Thirty-seven pediatric patients were studied with biplane ventricular cineangiography. There were 20 male and 17 female patients whose mean age at the time of the Fontan operation was 6.5 +/- 3.5 years (range 2.5 to 15.6). Eighteen patients were studied preoperatively, 25 at > 1 year postoperatively and 6 serially. Wall motion was assessed by a centerline method. Normal ranges for wall motion and other variables were established from 25 normal subjects. RESULTS: Wall motion abnormalities were observed in 2 of 18 patients preoperatively and in 11 of 25 patients postoperatively. Age at operation and ventricular volumes did not differ between postoperative patients who had normal (group I, 14 patients) or abnormal (group II, 11 patients) wall motion. However, ventricular mass and the mass/volume ratio were significantly greater and systolic variables and cardiac index were significantly lower in group II versus group I. Two patients in group I were considered to have a clinically poor outcome (persistent heart failure), and five in group II had heart failure, including one who died late. CONCLUSIONS: These observations suggest that postoperative regional wall motion abnormalities in this setting are not rare, may be related to excessive hypertrophy and may contribute to cardiac dysfunction and a poor clinical outcome. PMID- 8409059 TI - Cardiologic abnormalities in Noonan syndrome: phenotypic diagnosis and echocardiographic assessment of 118 patients. AB - OBJECTIVES: The purpose of this study was to determine the incidence of cardiologic abnormalities in Noonan syndrome. BACKGROUND: The incidence of cardiac abnormalities in Noonan syndrome remains unknown, largely because of such difficulties as assembling a substantial cohort, ensuring a correct phenotypic diagnosis and providing accurate definitions of the most frequent abnormalities- pulmonary stenosis and left ventricular hypertrophy. METHODS: A cohort of 145 patients was assembled, and before cardiologic assessment two independent geneticists scrutinized the phenotype. The diagnosis was confirmed in 118 patients, and they were studied by two-dimensional and Doppler echocardiography. RESULTS: A dysplastic pulmonary valve was present in eight patients (7%) and was associated with significant stenosis in six (75%) of the eight. Significant stenosis was present in 22 (20%) of 110 patients without dysplasia. Left ventricular hypertrophy was present in 29 patients (25%) without significant pulmonary stenosis. Localized anterior septal hypertrophy was the most common pattern in 12 (41%) of 29 patients. Diffuse hypertrophy involving the entire septum and the free wall was present in nine patients (31%) and was severe (> 1.7 cm) in five. Other abnormalities included secundum atrial septal defects (10%). CONCLUSIONS: The high incidence of cardiac abnormalities suggests that echocardiographic and Doppler evaluation of patients with the Noonan phenotype is important because it will aid in genetic counseling and in the assessment of the natural history of--and, ultimately, identification of the gene(s) responsible for--Noonan syndrome. PMID- 8409060 TI - Safety and efficacy of a new transpulmonary echo contrast agent in echocardiographic studies in patients. AB - OBJECTIVES: This study was designed to investigate in patients the effect of a new transpulmonary echo contrast agent, made from 5% human serum albumin (Albunex), on systemic and pulmonary hemodynamics and the influence of the contrast doses on left ventricular opacification. BACKGROUND: New intravenous transpulmonary echo contrast agents are promising, allowing contrast stress echocardiography and myocardial contrast echocardiography. Nevertheless, some shortcomings still remain. Thus, the pulmonary hypertension observed in pigs after Albunex injection should be investigated in humans, and the optimal dose of contrast agent remains to be determined because previous experiments indicated that the left ventricular opacification and attenuation are dose dependent. METHODS: Albunex in doses of 0.08 and 0.22 ml/kg was successively injected intravenously in 20 catheterized patients; in 11 of them, anti-inflammatory drugs were withdrawn to avoid the blocking of an eventual thromboxane-mediated pulmonary artery hypertension. Systemic blood pressure and pulmonary artery, capillary wedge and right atrial pressures were continuously monitored. Cardiac output, left ventricular fractional shortening and blood gases were determined 5 min before and 5 and 10 min after each injection. The left ventricular opacification was qualitatively assessed by three independent observers using a grading scale from 0 to 3, with 0 indicating an absence of contrast effect and 3 indicating full opacification. RESULTS: No clinical, hemodynamic or respiratory adverse reactions were observed in any patient. Irrespective of doses, a left ventricular opacification grade > or = 2 was observed in 74% of the 35 injections that could be evaluated. This percentage increased to 94% when the higher dose group was considered alone. CONCLUSIONS: This first report of the effect of Albunex injected intravenously on pulmonary artery pressures in humans demonstrates that this contrast agent appears to be safe. The significant left ventricular opacification obtained in a majority of patients without an important increase in attenuation supports the use of the higher dose of the contrast agent. PMID- 8409061 TI - Importance of electrode design, lead configuration and impedance for successful low energy transcatheter atrial defibrillation in dogs. AB - OBJECTIVES: We assessed the feasibility of low energy endocardial defibrillation in a canine model of atrial fibrillation, comparing catheters with large surface area electrodes and standard electrode catheters, and evaluated the effects of lead configuration and circuit impedance on defibrillation energy requirements. BACKGROUND: Although recent animal studies have demonstrated the feasibility of low energy endocardial atrial defibrillation, their results have been conflicting with regard to important methodologic aspects. METHODS: In 14 anesthetized greyhounds, atrial fibrillation was induced by rapid atrial pacing and maintained by vagal stimulation. Two large surface area braided electrode catheters and two standard electrode catheters were introduced percutaneously, one of each, in the right atrial appendage and right ventricular apex. A cutaneous patch electrode was placed on the left thorax. Biphasic shocks synchronized to the ventricular electrogram were used to terminate atrial fibrillation. Seven configurations were evaluated. Three used standard electrodes: proximal atrial cathode to distal atrial, ventricular or cutaneous anode. Four used braided electrodes: three with atrial cathode to ventricular, cutaneous or combined anode; one with ventricular cathode to atrial anode. RESULTS: Defibrillation with standard electrode catheters was associated with high impedance (576 +/- 112 omega) and low success rates for all configurations (28% success at < or = 40 J, no successes at 10 J). Low energy defibrillation was readily achieved with the braided electrodes with significantly lower impedance (75 +/- 13 omega, p < 0.0001). Ventricular fibrillation did not occur. The success rate of cardioversion increased in a dose response manner, allowing fitting of a sigmoid curve and calculation of energy associated with 50% (ED50) and 90% (ED90) success. The most successful configuration was ventricular cathode/atrial anode (ED50 1.5 +/- 0.4 J), and the least successful was atrial anode/cutaneous patch (ED50 6.5 +/- 3.2 J, p = 0.0001). CONCLUSIONS: Low energy atrial defibrillation is feasible using large surface area electrodes but not with standard electrode catheters owing to high impedance. An intracardiac anode provides lower impedance and higher success rates than are provided by a cutaneous anode. PMID- 8409062 TI - Implantation and recovery of temporary metallic stents in canine coronary arteries. AB - OBJECTIVES: The purpose of this study was to test the feasibility of implanting and retrieving a heat-activated recoverable temporary stent and to determine its effect on the angiographic, gross and histologic appearance of a normal coronary artery wall. BACKGROUND: Permanent coronary stenting is associated with a significant incidence of thrombosis, bleeding and vascular complications. These may be avoided by temporarily stenting for a period of hours to several days. METHODS: Seventy-eight stents constructed from the shape-memory nickel-titanium alloy nitinol were deployed by balloon expansion in the coronary arteries of 28 dogs and left in place for up to 6 months. Thirty minutes to 1 week after implantation, 70 stents were recovered by flushing the coronary arteries with 3 to 5 ml of 75 degrees C lactated Ringer solution, with collapse of the stent over a recovery catheter and subsequent withdrawal. RESULTS: All stents were successfully recovered and removed percutaneously. Mean vessel diameter after stenting was 12 +/- 6% (p < 0.05) greater than baseline diameter. Mean vessel diameter after stent removal remained enlarged (6 +/- 3%, p < 0.05). No angiographic or gross evidence of thrombosis, dissection, embolization, migration or spasm was associated with implantation or recovery. Microscopic examination revealed minor intimal injury in 40 segments (51%). Microscopic focal medial necrosis was associated with mural platelet-fibrin thrombus in 23 stented segments (29%) and media was interrupted in 7 (9%). CONCLUSIONS: This study demonstrates the feasibility of a new method of temporary stenting that uses the thermoelastic properties of nitinol to permit reliable recovery of the stent in normal canine coronary arteries. PMID- 8409063 TI - Reduction of ischemia-induced electrophysiologic abnormalities by glucose-insulin infusion. AB - OBJECTIVES: This study was designed to determine the effects of glucose-insulin infusion on ischemia-induced changes in extracellular potassium ([K+]o) accumulation and the associated electrophysiologic abnormalities in the canine heart. BACKGROUND: Although glucose-insulin-potassium infusion has been shown to limit myocardial injury in acute ischemia, its effect on ischemia-induced electrophysiologic alterations has not been investigated. METHODS: Recordings of [K+]o and local electrograms from the normal, border and ischemic zones were obtained during serial (10-min) left anterior descending coronary artery occlusions in the control state and after infusion of glucose-insulin (eight dogs), glucose alone (six dogs) or insulin alone (eight dogs). RESULTS: Glucose insulin infusion caused significant reduction in the rise of [K+]o during the entire period of ischemia in both ischemic and border zones associated with significant improvement in the degree of intramyocardial conduction delay. At 10 min of ischemia, [K+]o was reduced from a mean control level of 15.9 +/- 3.7 to 10.1 +/- 4.3 mmol/liter (p < 0.005) in the ischemic zone and from 6.8 +/- 1.9 to 5.5 +/- 1.1 mmol/liter (p < 0.05) in the border zone. The electrogram duration was shortened from a mean control value of 102 +/- 13 to 78 +/- 12 ms in the ischemic zone and from 79.2 +/- 7.8 to 58.1 +/- 6.6 ms in the border zone (p < 0.005). Glucose alone caused significant reduction in [K+]o during the initial 6 min of ischemia, only in the ischemic zone. Conversely, insulin caused no changes in [K+]o accumulation during ischemia. Neither glucose nor insulin alone had any effect on ischemia-induced intramyocardial conduction delay. CONCLUSIONS: The present study demonstrated that the combination of glucose and insulin is essential for the salutary effect of reducing [K+]o accumulation during ischemia and improving the associated intramyocardial conduction delay. It could be postulated that glucose in the presence of insulin increases the glycolytic flux, thereby providing adequate adenosine triphosphate for suppressing the cardiac adenosine triphosphate-sensitive potassium ion channels. The latter are, at least partially, responsible for the [K+]o rise in the early phase of ischemia. This study highlights the antiarrhythmic potential of interventions that modulate the metabolic consequences of ischemia. PMID- 8409064 TI - Sexual harassment and other gender issues in medicine. PMID- 8409065 TI - Education update: 1993. Training programs in the United States in adult cardiology, pediatric cardiology and cardiothoracic surgery. PMID- 8409066 TI - More thoughts on the nursing shortage. PMID- 8409067 TI - Antiphospholipid antibodies and valve disease in patients with systemic lupus erythematosus. PMID- 8409068 TI - Assessing collateral development after acute myocardial infarction. PMID- 8409069 TI - Randomized study assessing the effect of digoxin withdrawal in patients with mild to moderate chronic congestive heart failure: results of the PROVED trial. PROVED Investigative Group. AB - OBJECTIVES: The purpose of this study was to determine whether digoxin is effective in patients with chronic, stable mild to moderate heart failure. BACKGROUND: Digoxin has been a traditional therapy in heart failure, but methodologic limitations in earlier studies have prevented definitive conclusions regarding its efficacy. METHODS: Withdrawal of digoxin (placebo group, n = 46) or its continuation (digoxin group, n = 42) was performed in a prospective, randomized, double-blind, placebo-controlled multicenter trial of patients with chronic, stable mild to moderate heart failure secondary to left ventricular systolic dysfunction who had normal sinus rhythm and were receiving long-term treatment with diuretic drugs and digoxin. RESULTS: Patients withdrawn from digoxin therapy showed worsened maximal exercise capacity (median change in exercise time -96 s) compared with that of patients who continued to receive digoxin (change in exercise time +4.5 s) (p = 0.003). Patients withdrawn from digoxin therapy showed an increased incidence of treatment failures (p = 0.039) (39%, digoxin withdrawal group vs. 19%, digoxin maintenance group) and a decreased time to treatment failure (p = 0.037). In addition, patients who continued to receive digoxin had a lower body weight (p = 0.044) and heart rate (p = 0.003) and a higher left ventricular ejection fraction (p = 0.016). CONCLUSIONS: These data provide strong evidence of the clinical efficacy of digoxin in patients with normal sinus rhythm and mild to moderate chronic heart failure secondary to systolic dysfunction who are treated with diuretics. PMID- 8409070 TI - Sustained hemodynamic response to flosequinan in patients with heart failure receiving angiotensin-converting enzyme inhibitors. AB - OBJECTIVES: We evaluated the short- and long-term effects of flosequinan in 47 patients with severe heart failure despite ongoing captopril treatment. BACKGROUND: There have been no previous evaluations of the long-term hemodynamic effects of any direct-acting vasodilator in patients with heart failure receiving an angiotensin-converting enzyme inhibitor. Flosequinan is an arterial and venous vasodilator with actions similar to those of the hydralazine-isosorbide dinitrate combination. METHODS: After baseline hemodynamic measurements using balloon tipped pulmonary artery and radial arterial catheters, patients were randomized to receive 50, 100 or 150 mg of flosequinan daily. Hemodynamic variables were measured immediately before and after short-term flosequinan administration and after 8 weeks of therapy. RESULTS: With short-term flosequinan administration, mean arterial, right atrial and left ventricular filling pressures decreased by 6.4 +/- 1.1, 3.8 +/- 0.5 and 7.3 +/- 0.7 mm Hg, respectively (all p < 0.001). Cardiac index increased by 0.5 +/- 0.1 liters/min per m2, systemic vascular resistance decreased by 616 +/- 105 dynes.s.cm-5 and heart rate increased by 4 +/ 1 beats/min (all p < 0.001). After 8 weeks of long-term flosequinan administration, the vasodilator effect of a dose of flosequinan persisted. Compared with pretreatment baseline values, mean arterial, right atrial and left ventricular filling pressures at the peak effect of flosequinan were decreased by 3.5 +/- 1.3, 2.8 +/- 0.7 and 5.1 +/- 1.3 mm Hg, respectively (all p < 0.01). Systemic vascular resistance had decreased by 585 +/- 95 dynes.s.cm-5, cardiac index had increased by 0.5 +/- 0.1 liters/min per m2 and heart rate had increased by 10 +/- 2 beats/min (all p < 0.001). CONCLUSIONS: The arterial and venous vasodilator flosequinan exerts both short- and long-term sustained hemodynamic effects in patients with heart failure receiving angiotensin-converting enzyme inhibitors. PMID- 8409071 TI - Bedside cardiovascular examination in patients with severe chronic heart failure: importance of rest or inducible jugular venous distension. AB - OBJECTIVES: The aim of this study was to determine the sensitivity, specificity and utility of the cardiovascular examination in predicting cardiac hemodynamics in patients with advanced chronic congestive heart failure. BACKGROUND: Although the physical signs of acute left heart failure have been shown to correlate relatively well with cardiac hemodynamics, their reliability in estimating hemodynamics in patients with chronic heart failure has recently been questioned. METHODS: We prospectively recorded the history, cardiovascular physical signs present at bedside examination and the hemodynamic measurements obtained at right heart catheterization in 52 patients with chronic congestive heart failure undergoing in-hospital evaluation for possible heart transplantation. In addition, we obtained chest radiographs and multigated nuclear scans for the evaluation of left ventricular function. RESULTS: Pulmonary rales, a left ventricular third heart sound, jugular venous distension and the abdominojugular test, when positive, indicated higher right heart pressures and lower measures of cardiac performance. The presence of jugular venous distension, at rest or inducible, had the best combination of sensitivity (81%), specificity (80%) and predictive accuracy (81%) for elevation of the pulmonary capillary wedge pressure (> or = 18 mm Hg). Furthermore, in this population sample, the probability of an elevated wedge pressure was 0.86 when either variable was present. CONCLUSIONS: The bedside cardiovascular examination in the patient with chronic heart failure is extremely useful in identifying patients with elevation of right and left heart pressures. Examination for jugular venous distension at rest or by the abdominojugular test is simple and highly sensitive and specific in assessing left heart pressures in these patients. PMID- 8409072 TI - Effect on exercise performance of enalapril therapy initiated early after myocardial infarction. Nordic Enalapril exercise Trial. AB - OBJECTIVES: The Nordic Enalapril Exercise Trial was a multicenter subtrial of the Cooperative New Scandinavian Enalapril Survival Study (CONSENSUS II) designed to evaluate the effect on maximal exercise performance of a 6-month period of enalapril treatment initiated early after myocardial infarction. BACKGROUND: When begun early after myocardial infarction, converting enzyme inhibition therapy has been shown to attenuate infarct expansion and reduce left ventricular volume. Therapy has been associated with improved exercise performance. METHODS: Three hundred twenty-seven men (mean age 63.3 +/- 10.9 years) with documented acute myocardial infarction were randomized to treatment with enalapril or placebo on a double-blind basis. Intravenous enalaprilat or placebo therapy was initiated within 24 h after the onset of symptoms. Oral therapy was continued at a target dose of 20 mg/day. Patients exercised maximally at 1 month and 6 months after infarction to symptom-limited end points on a cycle ergometer with a 20 W/min incremental protocol. RESULTS: The treatment and control groups were comparable in patient age, concurrent therapy and type and site of infarction. At 1 month, for all patients, mean total work performed was 34.9 +/- 20.9 kJ in the enalapril group (n = 169) versus 28.5 +/- 20.6 kJ in the placebo group (n = 158) (difference = 18.4%, p < 0.01). This between-group difference in favor of enalapril was greatest in patients > 70 years old (difference = 41.4%, p < 0.01, n = 105) and those with clinical evidence of heart failure (difference = 33.0%, p < 0.01, n = 122). At 6 months for all patients, mean total work performed was 35.4 +/- 23.8 kJ in the enalapril group versus 34.0 +/- 23.9 kJ in the placebo group (difference = 4.1%, NS). CONCLUSIONS: This trial found that chronic converting enzyme inhibition initiated early after myocardial infarction was associated with significantly greater exercise capacity in men tested at 1 month. This difference was independent of type or site of infarction, patient age or the presence of clinical heart failure. The difference between the treatment and control groups was not significant at 6 months because of improvement in the placebo group. Further research is needed to elucidate the potential mechanisms involved, profile those patients most likely to profit from early therapy and establish the optimal timing and duration for intervention. PMID- 8409073 TI - Quantitative relation of myocardial infarct size and myocardial viability by positron emission tomography to left ventricular ejection fraction and 3-year mortality with and without revascularization. AB - OBJECTIVES: The purpose of this study was to determine the clinical prognostic value, with and without revascularization, of the size of myocardial infarction and viability as measured by positron emission tomography (PET). BACKGROUND: Poorly contracting but viable myocardium recovers contractile performance after revascularization. However, the quantitative relation among size of infarction and viability by PET, ejection fraction and long-term survival with and without revascularization in patients after myocardial infarction has not been previously reported. METHODS: Infarct size and viability imaged by PET using generator produced rubidium-82 were quantified objectively by automated software and related to coronary arteriography, left ventricular ejection fraction, revascularization and 3-year mortality. RESULTS: Myocardial infarction or scar > or = 23% of the left ventricle was associated with a 3-year mortality rate of 43% versus that of 5% associated with scar < 23% of the left ventricle (p = 0.014). An ejection fraction < or = 43% correlated with a 3-year mortality rate of 38% compared with 6% for an ejection fraction > or = 43% (p = 0.029) because infarct size > or = 23% of the left ventricle was also associated with an ejection fraction < or = 43%. For patients with a low ejection fraction (< or = 43%) or large infarcts/scar (> or = 23% of the left ventricle), ejection fraction value or infarct size did not predict mortality. However, in patients with an ejection fraction < or = 43%, the absence of viable myocardium in arterial zones at risk was associated with a mortality rate of 63% versus 13% in subjects with viable myocardium, a difference with only a 5.9% probability of occurring by chance alone (p = 0.059). For all patients with viable myocardium in arterial zones at risk, the mortality rate was 8%, and 80% had revascularization over 3 years. For patients with only fixed scar in arterial zones at risk, the mortality rate was 50% versus 8% (p = 0.018), and 40% had revascularization, with no difference in mortality with or without revascularization, thereby suggesting no benefit in this subgroup. CONCLUSIONS: Size of scar and viable myocardium by PET in arterial zones at risk in patients after myocardial infarction are highly predictive of 3 year mortality, particularly in patients with low ejection fraction, and identify patients who are suitable candidates for revascularization after myocardial infarction. PMID- 8409074 TI - Identification of a secondary peak in myocardial infarction onset 11 to 12 hours after awakening: the Cardiac Arrhythmia Suppression Trial (CAST) experience. AB - OBJECTIVES: The purpose of this study was to assess the relation between the time of awakening and the time of onset of acute myocardial infarction. BACKGROUND: Previous investigation has shown the onset of symptoms of acute myocardial infarction to have a primary peak 1 to 2 h after awakening. In studies not corrected for time of awakening, there appears to be a late afternoon/early evening peak, but data correlating the onset of symptoms with awakening have been limited by small numbers of patients, perhaps precluding identification of a secondary peak. METHODS: In the Cardiac Arrhythmia Suppression Trial (CAST), 3,549 patients had a documented myocardial infarction and entered antiarrhythmic drug titration. Of these, 3,309 had data on the onset of symptoms relative to the time of awakening and form the basis of this report. RESULTS: A total of 870 patients (26.3%) were awakened by symptoms. Of the remaining 2,439 patients who were not awakened by symptoms, 798 (32.7%) experienced the onset of symptoms in the 1st 4 h after awakening (with the highest number in the 1st h), after which the incidence of symptom onset decreased in a linear fashion, with a secondary peak 11 to 12 h after awakening. Both peaks are statistically significant. A similar pattern was seen in most of the subgroups examined (based on age, gender and various other demographic characteristics). CONCLUSIONS: Analysis of the very large CAST data base confirms the relation between awakening and onset of symptoms of myocardial infarction, suggesting involvement of the morning catecholamine surge. A secondary peak in symptom onset, occurring 11 to 12 h after awakening, is a new observation and may relate to ingestion of the evening meal or other trigger factors concentrated in those hours. PMID- 8409075 TI - A new nutritional approach in cancer therapy in light of mechanistic understanding of cancer causation and development. PMID- 8409076 TI - Hypothesis: dietary management may improve survival from nutritionally linked cancers based on analysis of representative cases. AB - A limited number of case histories was analyzed and verified to examine the effect of a very low fat, moderately high fiber, and moderately reduced calorie diet on the survival and quality of life of patients with primary cancer of the pancreas, metastatic stage D2 prostate cancer, and other nutritionally linked cancers. The retrospective study of pancreatic cancer patients disclosed that 1 year survival was higher among those who modified their diets than in those for whom there was no evidence as to diet alteration. For patients with metastatic prostate cancer (stage D2), a case control study demonstrated a statistical association of dietary modification with longer survival and improved quality of life. A retrospective study utilizing questionnaires supported such dietary modifications as a useful tool in the management of nutritionally linked cancers. PMID- 8409077 TI - Polysymptomatic syndromes and autonomic reactivity to nonfood stressors in individuals with self-reported adverse food reactions. AB - This study compared symptom reports and cardiovascular reactivity of a group of 24 individuals recruited from the community who reported a cognitive or emotional symptom caused by at least one food (food-sensitivity reporters, FSR) vs those of 15 controls (C) without a history of food, chemical, drug, or inhalant sensitivities. The main findings were: 1) FSR indicated sensitivities not only to foods, but also to environmental chemicals, drugs, and natural inhalants, as well as significantly more symptoms than C in multiple systems; 2) more FSR than C noted recent state depression and anxiety, as well as higher trait anxiety on the Bendig form of the Taylor Manifest Anxiety Scale; 3) however, on multiple regression analysis, not only depression, but also the number of sensitivities (foods, chemicals, drugs, inhalants), accounted for part of the variance in total number of symptoms (38 and 17%, respectively), whereas none of the affective measures accounted for any of the variance in total number of sensitivities over all subjects; 4) after controlling for depression and anxiety, FSR still showed a trend toward poorer performance on a timed mental arithmetic task (p = 0.16); and 5) FSR and C showed opposite patterns of heart rate change to two different stressful tasks (mental arithmetic and isometric exercise) (group by task interaction, p < 0.05). The data are discussed in terms of a time-dependent sensitization (TDS) process that predicts a cross-sensitizing and cross-reactive role for xenobiotic agents (e.g., foods, chemicals, drugs, and inhalants) and for salient psychological stress in the expression of psychophysiological dysfunctions of FSR. As in other chronically ill populations, negative affect in food-sensitive individuals may explain greater symptom reporting, but not necessarily account for the illness itself. For either a food or a psychological stimulus to begin to elicit sensitized responses, e.g., marked physiological differences from C, FSR may require multiple, intermittent exposures spaced over 5-28 days rather than on only 1 day. PMID- 8409078 TI - A review of persistent, low-grade lead challenge: neurological and cardiovascular consequences. AB - Although acute toxicity following heavy intake of lead (Pb) is a well-established clinical entity, the harmful effects of persistent, low-dose challenge, a situation commonly found among the general population, is uncertain. The major dangers of persistent, low-dose challenge that have been hypothesized are controversial: first, mentation and behavioral perturbations and second, development of hypertension with its consequences on the cardiovascular system. Accordingly, one cannot exclude some contributions from persistent Pb exposure to chronic disease and the aging process. Despite these potential adversities, many sources for Pb contact still remain with us. Current estimates are that 10-50% of American children (over 3-4 million) harbor unsafe levels according to present day standards. Therefore, it is reasonable to conclude that more work is needed in this area. Clearcut evidence concerning the deleterious influence of Pb on the nervous and/or cardiovascular-renal systems would lead to greater attempt to lessen exposure, to ameliorate symptomatology by providing supplemental agents which obviate the unwanted effects of Pb (iron, calcium, zinc), and to consider therapy with binding agents, like CaNa2EDTA, in the afflicted. PMID- 8409079 TI - Effect on mortality and reinfarction of adding fruits and vegetables to a prudent diet in the Indian experiment of infarct survival (IEIS). AB - The effects of antioxidant-rich foods as adjuncts to a prudent diet were compared for 12 weeks in a randomized, single-blind and controlled trial in 204 (group A) and 202 (group B) patients with acute myocardial infarction. There was a significant decrease in cardiac end points in group A compared to group B (37 vs 58, p < 0.01) after 12 weeks. Within intervention group A, those 108 patients with greater adherence to the intervention program showed a greater reduction in cardiac end points (14 vs 58, p < 0.001), and a significant decrease in total mortality (6 vs 28, p < 0.001), including cardiac mortality (6 vs 25, p < 0.01) compared to group B. Underlying these beneficial effects, antioxidant-rich foods caused a significantly smaller rise in lactate dehydrogenase (LDH) cardiac enzyme in group A than in group B (427.8 vs 561.6 IU/dL), indicating that the protective influence of such a diet may be observed within 1 week. The subset of group A patients showing reduction in mortality also had a lesser rise in LDH and greater reduction in blood lipids, blood glucose and blood pressures. Antioxidant-rich foods also caused a significant decrease in blood lipids with a lower decrease in high-density lipoprotein cholesterol in group A than in group B. Assay of serum level of antioxidants and long-term follow-up may confirm our observations. PMID- 8409080 TI - Blood pressure differences in older black and white long-term vegetarians and nonvegetarians. AB - The vegetarian diet has been associated with lower blood pressure (BP) in elderly white Americans. This study was undertaken to determine whether or not long-term adherence (at least 5 years) to a plant-based diet is similarly related to lower BP in older black Americans, a group exhibiting significant risk for hypertension (HT). Anthropometric characteristics, nutrient intake, and resting systolic and diastolic BP were measured in older black vegetarians (n = 27, age = 69.3 +/- 1.7 years), black nonvegetarians (n = 37, age = 65.4 +/- 1.2 years), white vegetarians (n = 85, age = 66.7 +/- 1.0 years), and white nonvegetarians (n = 54, age = 65.2 +/- 0.9 years). Older black vegetarians were significantly leaner and exhibited lower average systolic BP (131.4/76.8 mm Hg) and less hypertension than the black omnivores (141.6/76.2 mm Hg), but had significantly higher average BP than either dietary group of older white adults (vegetarians: 120.9/66.7 mm Hg; nonvegetarians: 122.8/67.6 mm Hg). These data suggest that long-term adherence to a vegetarian diet by older black Americans may afford some protection against hypertension, but in comparison to older white adults, does not completely offset their apparently greater susceptibility to untoward elevation of BP. PMID- 8409081 TI - Cholesterol screening in children: should obesity be a risk factor? AB - To determine whether obesity should be added to the current American Academy of Pediatrics (AAP) criteria for cholesterol screening in childhood, the charts of 99 children referred for evaluation of either hypercholesterolemia (n = 53) or obesity (n = 45) were reviewed. Compared with obese children, nonobese hypercholesterolemic subjects were younger (8.4 vs 11.4 years) and had lower mean body mass index and % ideal body weight. Frequency of elevated (> 90th percentile for age) total and low-density lipoprotein cholesterols were similar in both groups. Fifty-three of 65 children who met the current AAP criteria were hypercholesterolemic, however, 23/76 hypercholesterolemic children failed to satisfy these screening criteria. Thirty-six of 45 obese children had cholesterol levels > 90th percentile, suggesting increased risk for hypercholesterolemia in this group. If obesity was added to the AAP criteria, 66/80 hypercholesterolemic subjects would have been identified. These modified criteria, vs AAP standards, significantly improved both their sensitivity (70 vs 87%, p < 0.02) and negative predictive value (45 vs 30%, p < 0.02). Pending further studies in larger pediatric populations, these data indicate that obesity should be considered a risk factor for hypercholesterolemia in childhood, and we recommend modifying the AAP screening criteria to include obese children. PMID- 8409082 TI - Influence of dietary fat quantity and composition on insulin binding to rat intestine. AB - It is presently recommended that the general US population reduce the consumption of dietary lipid in order to reduce the risk of several chronic diseases, although the mechanism(s) through which dietary factors alter cellular function remain unclear. Dietary lipid composition has been shown to alter the plasma membrane lipid composition of adipocytes, muscle and other tissues. These changes in membrane lipid composition have been correlated with altered insulin receptor binding and signal transduction. Insulin receptors are present on mucosal cells of the intestinal tract, although their role in this tissue is not fully understood. We have fed rats diets containing 6, 31.4 or 76% of calories from lard (Protocol 1) and found insulin binding to be increased in the duodenum and decreased in the colon of rats fed the high-fat diet. Additionally, we compared diets containing either 12 or 37.6% of calories from beef tallow (saturated fatty acids or SFA) or corn oil (polyunsaturated fatty acids or PUFA; Protocol 2) and found insulin binding in the jejunum to be significantly decreased by a low SFA or high PUFA diet relative to the low PUFA diet. These results suggest that intestinal insulin receptors are responsive to dietary lipid quantity and quality which may have implications as to the role of dietary factors in modifying nutrient transport and/or risk of intestinal disease. PMID- 8409083 TI - Home parenteral nutrition after near total enterectomy. AB - Six patients with extreme short bowel syndrome (4.2 +/- 4.9 cm of residual small bowel) were provided home parenteral nutrition (HPN) for 14,397 days. The average age at onset of HPN was 38 years (18-64 years). Patients maintained body weight at 97% of ideal (86-112%) with mean serum albumin of 3.7 +/- 0.6 g/dL (normal 3.5 5.8 g/dL), serum transferrin of 341 +/- 104 mg/dL (normal 200-400 mg/dL), and mean serum pre-albumin of 27.5 +/- 12.6 mg/dL (normal 16.6-43 mg/dL). Hospital admission for HPN-related complications was required 10.3 times/patient for a total of 864 hospital days and was catheter related in 71% of episodes. Catheter sepsis occurred once per 436 catheter days and required catheter removal in only 33% of instances. Five patients were able to resume an oral diet, five had returned to work or school, and three were married with family. HPN allowed return to a quality productive life with acceptable morbidity following catastrophic massive bowel resection. PMID- 8409084 TI - Responses of plasma magnesium and other cations to fluid replacement during exercise. AB - Changes in plasma magnesium (Mg) and other cations in response to exercise have been previously reported, but whether ingestion of fluids providing carbohydrate and electrolytes modifies the changes has not been determined. We examined patterns of change in plasma Mg, calcium (Ca), sodium (Na), and potassium (K) in 10 men [age 30 +/- 2 years; maximal oxygen uptake (VO2max) 57.4 +/- 3.2 ml.kg 1.min-1] who ran on a treadmill for 2 hours at 60-65% of their VO2max. Subjects drank 200 mL of water (W) or a 7% glucose polymer/fructose/electrolyte solution (GPFE) at 0 time and every 30 minutes while running (30, 60 and 90 minutes). Plasma Mg continued to decline throughout exercise and was lowest when exercise was terminated at 120 minutes (87.9 +/- 1.2 and 88.9 +/- 2.2% of pre-exercise for W and GPFE, respectively). In contrast, a significant increase in serum K was noted (105.2 +/- 3.6 and 112.8 +/- 3.0% of pre-exercise at time 120 for W and GPFE). Serum Na increased slightly, but no changes in plasma Ca were observed. Serum glucose and lactate increased transiently for all running conditions, but no differences across fluid treatments were noted. Serum free fatty acids (FFA) increased during exercise with W ingestion, whereas no rise in serum FFA occurred with GPFE until the end of exercise. That patterns of change in Mg and K during exercise were not altered by providing a fluid replacement beverage containing glucose polymer, fructose, Mg, K, Ca, and Na, as compared to water, suggests that these events may be requisite to maintenance of homeostasis. Mechanisms must be sought to explain the decline in plasma Mg that occurs with exercise. PMID- 8409085 TI - Molecular biological approaches to studying trace minerals: why should clinicians care? AB - The approaches and tools of molecular biology have been enormously valuable to all branches of biological science over the last decade. Nutrition is no exception, where studies on the influence of nutrients on gene expression and of gene products on nutrient metabolism have resulted in a much more sophisticated and detailed understanding of nutritional biochemistry. An example of this as applied to trace minerals research can be seen in the area of thyroidology. Until recently, the sole link between thyroid hormones and trace minerals was iodide. Then the thyroid hormone receptor was cloned and analysis of the protein coding sequence showed it to be a member of a large family of gene activating receptor proteins. These all possess a region containing two clusters of cysteine residues, thought to chelate zinc, which is required for binding of the receptors to their target genes. Zinc appears to be necessary for the biological functioning of not only the thyroid hormone receptor but also many other nuclear proteins which regulate gene expression. The principal product of the thyroid gland is thyroxine from which the more active form of the hormone, triiodothyronine, is derived by peripheral monodeiodination. One of the two enzymes responsible, type I 5'-iodothyronine deiodinase, was recently cloned and shown to contain selenocysteine. Thus production of the active thyroid hormone is dependent on selenium status. These advances made with molecular biology have important implications for clinicians. The possibilities for understanding the clinical picture are immediately enhanced, improving both diagnosis and treatment. Molecular biology also provides the opportunity for developing more specific and sensitive tools for assessing nutritional status. Diseases with a genetic basis can be unequivocally diagnosed and perhaps even treated. A strength of nutrition is that it encompasses molecular biology and clinical practice and practitioners of each can benefit from an understanding of the complementary area. PMID- 8409086 TI - Copper deficiency in rats: the effect of type of dietary protein. AB - The present investigation was conducted to determine whether type of dietary protein can exacerbate the pathology induced by the combination of fructose feeding and copper (Cu) deficiency. Weanling male Sprague-Dawley rats were assigned to three different groups differing in the nature of dietary protein. The proteins used were egg-white, casein or lactalbumin. All diets contained 62.5% carbohydrate as fructose and were low in Cu (0.6-0.72 microgram Cu/g diet). Although the lowest concentration of Cu was found in the livers of rats fed egg white, the pathology associated with Cu deficiency was more severe in rats fed lactalbumin. The highest concentration of hepatic Cu was found in rats fed casein. The data show that the type of dietary protein can exacerbate signs associated with Cu deficiency. The concentrations of hepatic Cu do not reflect accurately the pathology associated with Cu deficiency. PMID- 8409087 TI - Dysprosium as a nonabsorbable marker for studies of mineral absorption with stable isotope tracers in human subjects. AB - Two studies were conducted to determine if dysprosium (Dy) could be used as a quantitative fecal marker for studies of zinc-70 (70Zn), copper-65 (65Cu) and magnesium-26 (26Mg) absorption in humans. In the first experiment, Dy excretion was shown to be complete (104 +/- 9%; mean +/- SD, n = 6) and the kinetics of fecal Dy excretion closely paralleled that of 70Zn but not 65Cu. Because of the similarity in 70Zn and Dy excretion kinetics, a method for estimating 70Zn absorption was developed which used 70Zn and Dy data from only the first two stools passed after isotope administration. Average estimates of 70Zn absorption based on the two-stool (partial pool) vs total pool (5-day composite) method were not different (28.0 +/- 5.2 vs 24.4 +/- 4.1%, respectively; means +/- SEM; p > 0.10). In the second study, the same questions was addressed relative to 26Mg absorption. 26Mg and Dy also exhibited nearly identical excretion patterns. Average estimates of 26Mg absorption based on the partial pool vs total pool method were comparable yielding mean values of 22.7 +/- 3.4 vs 23.2 +/- 2.5% and 26.1 +/- 1.8 vs 24.3 +/- 1.8%, respectively, from magnesium-26 oxide (26MgO) and 26Mg glycinate. Advantages and limitations of the partial pool method for estimating mineral absorption are discussed. PMID- 8409088 TI - Biopotencies of SRR- and all-rac-alpha-tocopheryl acetate are significantly different. PMID- 8409089 TI - What happens to blindly placed nasoenteral tubes? PMID- 8409090 TI - Women's health and nutrition issues. PMID- 8409091 TI - Women's health and nutrition research: US governmental concerns. AB - The US Public Health Service (USPHS) and the National Institutes of Health (NIH) have a major commitment to women's health issues and to women's health research. "To assess the problems of women's health in the context of the lives women in America lead today," the Assistant Secretary for Health, USPHS, appointed a task force whose report published in 1985 identified issues, listed 15 recommendations, and served as a guide for the establishment of groups within each agency to implement the recommendations according to their appropriate responsibilities. NIH established an Advisory Committee on Women's Health Issues which assessed NIH involvement in women's health research, made recommendations for implementation or expansion of this research, including a recommendation that women be included in clinical trails or their exclusion be justified. An Office of Research on Women's Health was created. The NIH announced a Women's Health Initiative (WHI) in the spring of 1991, which will address three of the leading health problems for women: cardiovascular disease, breast and colon cancer, and osteoporosis. The WHI will provide an integrated, multidisciplinary approach through clinical trials, observational studies, and community trials. Clinical trials will evaluate hormone replacement therapy, calcium/vitamin D, and dietary modification of fat/fiber. Community trials will implement known interventions for relevant risk factors. Presented here are USPHS response to the mandate to address women's health issues; data from the US National Center for Health Statistics about the incidence and prevalence of diseases and risk factors in women; details about the WHI clinical trial; and aspects of the WHI applicable to the field of nutrition. PMID- 8409092 TI - Nutrition and the older female: a review. AB - Reviewed here are the major nutritional problems of older females, stressing where older female nutrition differs from that of older males. Older females have a higher body mass index and lower waist-to-hip ratio than do older males. Older females reduce their food intake compared to younger females much less than do older males compared to younger males. Total energy expenditure is minimally reduced in older females compared to young females. Resting metabolic rate is reduced by 13% in older females. The major nutritional problem faced by older females, protein energy malnutrition, and its reversible causes are addressed. Inability to recognize the need for fluids is a common problem leading to dehydration in older persons. Older persons are at major risk for ingesting less than two-thirds of the recommended dietary allowance for vitamins. Older women are more likely than men to take vitamin or mineral supplements. Osteopenia is a common problem leading to fractures in older women, and decreased calcium intake and vitamin D deficiency, as well as estrogen deficiency, play a role in the pathogenesis of osteopenia. Total cholesterol levels increase with age in older women, while high-density lipoprotein cholesterol levels decline. However, cholesterol levels that are optimum for survival are higher in older women than in older men. At < 65 years of age, diabetes mellitus (DM) is more common in females, and at > 65 years of age it is more common in males. Over one-half of older persons with DM are undiagnosed. Management of DM in older persons requires a judicious approach to balance the potential problems of hypo- and hyperglycemia. PMID- 8409093 TI - Nutrition issues of military women. AB - Nutritional data were collected from 147 female soldiers in four surveys (two field and two dining hall studies) conducted by the US Army Research Institute of Environmental Medicine and Letterman Army Institute of Research. The 49 female soldiers observed in the field studies had suboptimal mean energy (1751 kcal), protein (52.3 g), calcium (620 mg), and iron (11.8 mg) intakes. In addition, 54 female soldiers in the dining hall studies also had lower than desirable mean iron (16.2 mg) intakes. These data suggest that nutritional problems encountered by military women are similar to those faced by their civilian counterparts, based on NHANES II and CSFII reports. Although the impact of sporadic low nutrient intakes during short 10-14-day field training exercises may be of little consequence, this may be significant when inadequate intakes occur routinely or for extended periods during military conflicts. Viable solutions to the nutritional problems of military women could include ration supplementation and nutrition education. PMID- 8409094 TI - Alcohol and nutrition in postmenopausal women. AB - The estrogenization of postmenopausal women is of major importance for their health status, particularly with respect to risks of osteoporosis and coronary heart disease. Thus the factors which influence endogenous postmenopausal estrogen levels are receiving increasing attention. There are three major determinants of endogenous estrogen levels; two are well established, while the third is of fairly recent vintage. Two of the three are nutritionally based. A long-recognized nutritional determinant is body fat mass; a newly recognized determinant is that of moderate alcoholic beverage consumption. Another recognized but non-nutritional postmenopausal estrogen determinant is the presence of the ovaries. This review will examine these three determinants of endogenous postmenopausal estrogen levels. Further, data will also be presented to indicate that nationality may be a fourth factor, and may involve a potential nutritional component. Finally, to explore more deeply the nutritional aspects of alcoholic beverage consumption, the effects of phytoestrogen congeners of alcoholic beverages on the estrogenization of postmenopausal women will be reviewed. PMID- 8409095 TI - Healthy adiposity in women: the Framingham Offspring Study. AB - This report estimates a threshold for "healthy adiposity" by determining the relationship between subscapular skinfolds thickness, a direct measure of adiposity, and cardiovascular disease risk factors in 1254 nonsmoking adult women who participated in the Framingham Offspring Study. Cardiovascular disease risk factors, including blood pressure, fasting plasma lipoprotein cholesterol, plasma glucose and echocardiographically measured left ventricular mass, were included. The optimal subscapular skinfold for 20-39-year-old women was determined to be < 15 mm. Women aged 20-59 whose subscapular skinfolds were below this level had a mean body mass index (BMI) of 21.1 kg/M2. Weight-for-height estimates for these women corresponded closely to the 1959 Metropolitan Life Insurance Company Desirable Weight table. The probability of having unhealthy adiposity was estimated for all women aged 20-59 for each (rounded) integer value grouping of BMI. The probability of being above the healthy adiposity threshold rises rapidly across BMI levels > 20 and plateaus > 24. Thus, only women with BMI < 24 need assessment for adiposity status. PMID- 8409096 TI - Exercise is not an effective weight loss modality in women. AB - The excess caloric expenditure which results from physical activity should lead to weight loss if caloric expenditure at other times remains constant. Unfortunately, while there is good evidence for such an effect in men, there is little if any evidence for a similar effect in women. Weight loss with exercise does not readily occur in women unless accompanied by caloric restriction. Further, the role of exercise in maintaining resting metabolic rate while dieting has only marginal support. Potential reasons for the ineffectiveness of exercise in inducing weight loss in women include smaller body size and lower aerobic capacity, under-reporting of caloric intake, differences in body fat distribution and sensitivity to catecholamines, a different gonadal hormone milieu, and energy conservation resulting from evolutionary pressures. Nevertheless, regular exercise in women has many beneficial effects on lipids, glucose homeostasis and bone metabolism even if weight loss does not occur. PMID- 8409097 TI - Physical activity, physical fitness, and all-cause mortality in women: do women need to be active? AB - Physical inactivity is associated with higher mortality rates in most studies in men, but studies in women are more equivocal. The purpose of this study was to evaluate the relationship of sedentary living habits to all-cause mortality in women. A group of 3,120 adult women completed a preventive medical examination, and were followed for approximately 8 years for mortality. There were 43 deaths and a total of 25,433 person-years observed during follow-up. Physical fitness was assessed at baseline by a maximal exercise test on a treadmill, and physical activity was estimated by a self-administered questionnaire. Age-adjusted all cause mortality rates were significantly inversely associated with physical fitness. Death rates were 40, 16, and 7 per 10,000 person-years of follow-up across low, moderate, and high categories of physical fitness, respectively. However, death rates did not differ across low, moderate, and high categories of physical activity. These findings are different than for men in the same study, where both physical activity and physical fitness were inversely associated with mortality risk. We attribute the lack of association between physical activity and mortality in women to be due to inadequate assessment of activity, and that this also is the likely explanation for the difference in results between women and men in published studies of physical activity and mortality. PMID- 8409098 TI - Effects of gestation, lactation, and maternal calcium intake on mechanical strength of equine bone. AB - Skeletal homeostasis during late gestation, lactation, and the post-lactational recovery period is poorly understood. In an experiment using an animal model (the horse), metacarpal breaking strengths (MBS) estimated via transmission ultrasonics were examined during the last 12 weeks of gestation and for 40 weeks after parturition. MBS increased during the last 6-10 weeks of gestation in mares fed amounts of calcium (Ca) recommended by the National Research Council; maximum MBS coincided with the week of parturition. In contrast, MBS in mares fed 20% less Ca remained relatively constant during the last 12 weeks of gestation. In contrast to increases during late gestation, MBS decreased steadily in all mares during the first 12 weeks of lactation. MBS increased after approximately 12 weeks of lactation, but more slowly than they had declined. MBS of the bones of mares fed recommended amounts of Ca were fully restored at 24 weeks post parturition, but those of Ca-deficient mares had not fully recovered even 20 weeks after milk production had ceased (40 weeks after parturition). Mid-cannon mediolateral diameters of foals born to mares fed Ca-deficient diets were thinner and mechanically weaker at birth (both p < 0.01). These differences in limb bone size and strength persisted during the first 40 weeks of life. PMID- 8409099 TI - Contributions of dietary calcium and physical activity to primary prevention of osteoporosis in females. AB - In the United States and many Western nations, increasing prevalence of osteoporotic fracture is contributing to the health care system burden, and costs and needs for medical services are predicted to increase significantly by the beginning of the 21st century. Recent reports investigating developmental changes in skeletal mass of adolescent girls and young women under different experimental or ecological conditions support the contention that modification of environmental factors, especially dietary calcium and physical activity, can favorably modulate bone mass and bone density compared to controls. The peripubertal period, starting as early as 10 years of age, seems to be most responsive to modification of environmental/lifestyle factors, whereas potential gains of bone mass during late adolescence and early adulthood, although smaller, may be more readily achieved through improved dietary calcium intakes and regular exercise programs. Scientific evidence in support of these beneficial effects on bone is presented as part of the rationale for prevention of osteoporotic fractures. Also discussed is the need for a national policy to prevent osteoporosis through primary prevention strategies focused at young girls prior to puberty. The goal would be for every prepubertal girl, over the next decade, to achieve peak bone mass (and density) of the spine, hips and other bones by age 20, i.e., early adulthood. A second target group for national policy should be women in their 20s. Attainment of the goal to gain 3-5% additional bone mass during this decade would prolong the time before the fracture threshold range (low bone mass) is reached in the postmenopausal decades. PMID- 8409100 TI - The role of trace minerals in osteoporosis. AB - Osteoporosis is a multifactorial disease with dimensions of genetics, endocrine function, exercise and nutritional considerations. Of particular considerations are calcium (Ca) status, Vitamin D, fluoride, magnesium and other trace elements. Several trace elements, particularly copper (Cu), manganese (Mn) and zinc (Zn), are essential in bone metabolism as cofactors for specific enzymes. Our investigations regarding the role of Cu, Mn and Zn in bone metabolism include data from studies with animals on Cu- and Mn-deficient diets. We have also demonstrated cellular deficiencies using bone powder implants, as well as fundamental changes in organic matrix constituents. In clinical studies we have demonstrated the efficacy of Ca, Cu, Mn and Zn supplementation on spinal bone mineral density in postmenopausal women. Each of these studies demonstrated the necessity of trace elements for optimal bone matrix development and bone density sustenance. PMID- 8409101 TI - Dietary fat, calories, and the risk of breast cancer in postmenopausal women: a prospective population-based study. AB - We tested the hypothesis that a high-fat diet increases the risk of breast cancer in a population-based study of 590 women aged 40-79 years who were without known breast cancer when they provided a quantitative 24-hour diet recall. Fifteen postmenopausal women were diagnosed with incident breast cancer during the next 15 years (approximately 7600 person-years of follow-up). These women had significantly higher age-adjusted intake of all fats (monounsaturated, polyunsaturated, and saturated), and oleic, linoleic, and linolenic acids, with a stepwise increase in risk across tertiles of intake. Fat intake was associated with total calories, protein, and carbohydrates, and women with incident breast cancer consumed more calories, protein, and carbohydrates than did other subjects. When each nutrient variable (calories, fats, protein, and carbohydrates) was adjusted for age, body mass index, age at menopause, parity, and alcohol consumption, the strongest risks for incident breast cancer were associated with total calories (relative risk per standard deviation = 2.72, 95% confidence interval = 1.51-4.89, p = 0.002) and total fats (relative risk per standard deviation = 2.01, 95% confidence interval = 1.19-3.41, p = 0.01). Fat composition of the diet, expressed either as percent of energy or as fat intake adjusted for calories by regression analysis, was not significantly associated with risk of breast cancer. These results support the hypothesis that total calorie consumption, as well as dietary fat consumption, is a risk factor for breast cancer in postmenopausal women, and parallel observations in animal models. PMID- 8409102 TI - Antioxidant micronutrients and breast cancer. AB - We reviewed epidemiologic evidence on the relationship between four antioxidant micronutrients (vitamin A, vitamin C, vitamin E, and selenium) and breast cancer risk. Available data support a modest protective effect of vitamin A, although more studies are needed to examine further this association and to assess the relative contributions of preformed vitamin A (retinol) and carotenoids. In addition, the possibility that some other component of vitamin A-rich foods may account for this observed association should be explored. Data on the relationship between vitamins C and E and breast cancer risk are limited and inconsistent, and further information is necessary. A substantial body of evidence indicates a lack of any appreciable effect of selenium intake on breast cancer risk, at least within the range of human diets. Future observational studies should ideally be prospective in design, as prospective studies are less prone to selection and recall bias than are case-control studies, and should address methodologic issues such as confounding by other micronutrients and appropriate storage conditions of blood specimens. Although hypotheses relating micronutrient intake to risk of breast cancer should be tested in randomized trials, ethical and logistical constraints make these studies difficult to perform. PMID- 8409103 TI - Niacin deficiency and cancer in women. AB - A new interest in the relationship between niacin and cancer has evolved from the discovery that the principal form of this vitamin, NAD, is consumed as a substrate in ADP-ribose transfer reactions. Poly(ADP-ribose) polymerase, an enzyme activated by DNA strand breaks, is the ADP-ribosyltransferase of greatest interest with regard to effects on the niacin status of cells since its Km for NAD is high, and its activity can deplete NAD. Studies of the consequences of DNA damage in cultured mouse and human cells as a function of niacin status have supported the hypothesis that niacin may be a protective factor that limits carcinogenic events. To test this hypothesis in humans, we used a biochemical method based on the observation that as niacin nutriture decreases, NAD readily declines and NADP remains relatively constant. This has been demonstrated in both fibroblasts and in whole blood from humans. Thus, we use "niacin number," (NAD/NAD+NADP) x 100% from whole blood, as a measure of niacin status. Healthy control subjects showed a mean niacin number of 62.8 +/- 3.0 compared to 64.0 for individuals on a niacin-controlled diet. Analyses of women in the Malmo Diet and Cancer Study showed a mean niacin number of 60.4 with a range of 44 to 75. The distribution of niacin status in this population was nongaussian, with an unpredictably large number of individuals having low values. PMID- 8409104 TI - Nutrition and diseases of women: cardiovascular disorders. AB - Studies of prevention, diagnosis, and intervention for coronary heart disease and hypertension have either been conducted largely in men, or gender differences have not always been fully sought. This has added to a general perception that coronary artery disease is basically a male affliction despite the fact that coronary artery disease is the leading cause of death among women, especially elderly women. Many risk factors among women are similar to men, i.e., high blood pressure (BP), elevated serum cholesterol levels, and cigarette smoking; however, women compared to men have greater incidence of diabetes mellitus, congestive heart failure, and hypertension as they become older. The risk of cardiovascular disorders can be reduced by postmenopausal estrogen replacement, exercising sufficiently, and ceasing smoking. In addition, good nutrition, taking into consideration the proper amount and forms of calories, sodium, potassium, calcium, magnesium, and macronutrients to maintain an ideal lipid profile and BP, is helpful in preventing cardiovascular perturbations. PMID- 8409105 TI - Antioxidants and cardiovascular disease: a review. AB - In spite of the significant decline in cardiovascular disease (CVD) mortality over the past several decades, CVD remains the leading cause of death in the United States. Although age-specific CVD rates are higher in men than women, CVD is nonetheless the leading cause of death for both sexes, and is responsible for approximately one-third of all fatalities in women as well as men. Antioxidant vitamins are a promising area of current research in the prevention of CVD. The postulated mechanism for such an effect derives from basic research demonstrating the ability of antioxidants to inhibit the oxidation of low-density lipoprotein cholesterol. Epidemiologic studies that have explored the antioxidant vitamin hypothesis include descriptive and cross-sectional studies, analytic investigations using case-control and prospective cohort study designs, as well as several small randomized clinical trials. Findings from these studies are not totally consistent, but generally support the hypothesis that antioxidant vitamins reduce the risk of CVD. Overall, there are fewer data in women than men. Large-scale randomized trials are now ongoing that will provide reliable evidence on this question. The ongoing Physicians' Health Study of over 22,000 men is testing beta-carotene, while the recently begun Women's Health Study of 40,000 women will test, utilizing a factorial design, beta-carotene as well as vitamin E. A trial has also recently been funded to test beta-carotene, vitamin E and vitamin C in secondary prevention among a high-risk population of 8,000 women with prior CVD events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409106 TI - Diet, lipids and cardiovascular disease in women. AB - While hypercholesterolemia is an important risk factor for coronary heart disease in women, the responses of women and men to low-fat, low-cholesterol diets have not been carefully compared. This report summarizes three studies comparing responses of women and men. First, we evaluated the responses of women and men to public cholesterol screening and enrollment in a nutrition research study. Of the 3401 individuals who presented for initial screening, 56% were women and 44% were men. Of the eligible subjects, women (56%) were less likely than men (66%) to come to a second screening. After the second screening, however, study-eligible women (75%) were slightly more likely than men (67%) to enroll in the study. Next, serum and low-density lipoprotein cholesterol (LDL-C) values were compared for 64 women and 99 men in response to an American Heart Association (AHA) diet. Reductions in serum cholesterol (8.8 and 7.3%) and LDL-C (9.4 and 8.8%) were similar in women and men, respectively. Finally, serum lipid responses of 59 women and 87 men to a 12-month clinical trial with an AHA diet and a high-fiber AHA-type diet were compared. Both diets decreased serum cholesterol (10 and 13%) and LDL-C (14 and 18%), respectively, for the combined group of women and men. There were no significant differences between serum lipid responses for women and men. These observations suggest differences between the responses of women and men to screening and enrollment in a research study. However, responses of women and men to cholesterol-reducing diets appear similar.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409107 TI - Interrelationship of magnesium and estrogen in cardiovascular and bone disorders, eclampsia, migraine and premenstrual syndrome. AB - The anticonvulsive and antihypertensive values of magnesium (Mg) in eclampsia, and its antiarrhythmic applications in a variety of cardiac diseases, have caused Mg to be considered only for parenteral administration by many physicians. In contrast, nutritionists have long recognized Mg as an essential nutrient, because severe deficiencies elicit neuromuscular manifestations similar to those justifying its use in eclampsia. More recently, this element has been used to favorably influence latent tetany with and without thrombotic complications, to delay preterm birth, to influence premenstrual syndrome, and to ameliorate migraine headaches. Most of these disorders exclusively or largely afflict women. The lesions of arteries and heart caused by experimental Mg deficiency have been well documented and may contribute to human cardiovascular disease. Estrogen's enhancement of Mg utilization and uptake by soft tissues and bone may explain resistance of young women to heart disease and osteoporosis, as well as increased prevalence of these diseases when estrogen secretion ceases. However, estrogen induced shifts of Mg can be deleterious when estrogen levels are high and Mg intake is suboptimal. The resultant lowering of blood Mg can increase the Ca/Mg ratio, thus favoring coagulation. With Ca supplementation in the face of commonly low Mg intake, risk of thrombosis increases. PMID- 8409108 TI - A multivariate analysis of psychological factors related to body mass index and eating preoccupation in female college students. AB - Using a composite questionnaire, we measured the degree to which preoccupation with eating and body mass index (BMI) are related to a series of psychological measures in a sample of female college students. Eating preoccupation was measured using a set of questions designed to be free of affective content. Thirty volunteers filled out the questionnaire; their height and weight were measured under standard conditions. Correlations showed that BMI was positively related to the variables Depression and Emotional Response to Environmental Stimuli, while Eating Preoccupation was most closely related to Anxiety. Principal components analysis revealed two factors, one containing measures of positive emotionality, and one containing Eating Preoccupation and measures of negative emotionality, with BMI loaded on both components. These factors reflect the relationship of BMI to measures of both positive and negative emotionality. Positive emotionality may be a reflection of activation of a neurobehavioral system, the behavioral facilitation system, while negative emotionality may be a reflection of activation of the behavioral inhibition system. The overweight women in this study appeared to have high levels of activation of one or both systems, while the underweight women appeared to have low activation of both systems. Approaches to the study of obesity in women should take into account the fact that positive emotionality, as well as negative emotions such as depression and anxiety, may be related to BMI, and that eating preoccupation appears to be a subset of overall anxiety. PMID- 8409109 TI - Gastrointestinal and nutritional aspects of eating disorders. AB - Anorexia nervosa (AN) and bulimia nervosa (BN) are potentially fatal eating disorders which primarily affect adolescent females. Differentiating eating disorders from primary gastrointestinal (GI) disease may be difficult. GI disorders are common in eating disorder patients, symptomatic complaints being seen in over half. Moreover, many GI diseases sometimes resemble eating disorders. Inflammatory bowel disease, acid peptic diseases, and intestinal motility disorders such as achalasia may mimic eating disorders. However, it is usually possible to distinguish these by applying the diagnostic criteria for eating disorders and by obtaining common biochemical tests. The primary features of AN are profound weight loss due to self starvation and body image distortion; BN is characterized by binge eating and self purging of ingested food by vomiting or laxative abuse. GI complications in eating disorders are common. Recurrent emesis in BN is associated with dental abnormalities, parotid enlargement, and electrolyte disturbances including metabolic alkalosis. Hyperamylasemia of salivary origin is regularly seen, but may lead do an erroneous diagnosis of pancreatitis. Despite the weight loss often seen in eating disorders, serum albumin, cholesterol, and carotene are usually normal. However, serum levels of trace metals such as zinc and copper often are depressed, and hypophosphatemia can occur during refeeding. Patients with eating disorders frequently have gastric emptying abnormalities, causing bloating, postprandial fullness, and vomiting. This usually improves with refeeding, but sometimes treatment with pro motility agents such as metoclopromide is necessary. Knowledge of the GI manifestations of eating disorders, and a high index of suspicion for one condition masquerading as the other, are required for the correct diagnosis and management of these patients. PMID- 8409110 TI - Serum insulin-like growth factor-I concentrations in the recovery of patients with anorexia nervosa. AB - Anorexia nervosa (AN) can result in extreme malnutrition, and these patients frequently require inordinately large amounts of calories to gain weight during refeeding therapy. Insulin-like growth factor-I (IGF-I) is a polypeptide that mediates many of the anabolic effects of growth hormone. Low levels of IGF-I have been associated with malnutrition and can cause poor weight gain. To clarify the potential relationship of IGF-I to weight gain, serial serum IGF-I, retinol binding protein and prealbumin levels were measured at admission, 2 weeks and 4 weeks, in 14 consecutive consenting patients admitted for treatment of AN. Baseline IGF-I levels were lower in the patients compared to age-matched controls (mean 20.8 +/- 2.5 vs 32.9 +/- 2.9 nmol/L, p < 0.01). In patients with no weight gain, IGF-I levels were static. There was a stepwise increment in the IGF-I values related to weight gain. Retinol-binding protein and prealbumin, proteins commonly used to assess nutritional status, did not demonstrate important correlations with weight gain. Further studies are required to determine whether or not initial low IGF-I levels impede weight gain in AN patients and whether treatment with IGF-I (possibly in combination with growth hormone) may be of benefit in this disease process. PMID- 8409111 TI - Trends in asthma and chronic obstructive pulmonary disease therapy in Canada, 1985 to 1990. AB - BACKGROUND: In the mid and late 1980s, numerous changes were recommended in the management of obstructive lung diseases. We analyzed drug sales to determine whether these recommendations have resulted in recent changes in prescription drugs used in airway management. METHODS: Data on prescription sales in Canada from 1985 to 1990 were obtained from an international pharmaceutical market research organization. Data from a random sampling of physicians that reported patients seen along with the diagnosis and prescriptions given were also reviewed. RESULTS: From 1985 to 1990, there was a 38% increase in prescriptions of all airway drugs. Prescriptions increased significantly for inhaled drugs as follows: inhaled beta 2-agonists, 70%; inhaled steroids, 139%; cromoglycate, 88%; and ipratropium bromide, 204%. Theophylline prescriptions, by contrast, fell significantly, by 19%. Although theophylline was the most commonly prescribed airway medication in 1985, inhaled beta 2-agonists were the most commonly prescribed drug in 1990, and these changes in proportional use were significant. Increases in ipratropium bromide usage was accounted for by increases in prescriptions for chronic obstructive pulmonary disease. Increases in nonsteroidal antiinflammatory drugs were accounted for by asthma prescriptions. CONCLUSIONS: The increase in airway medication prescriptions dispensed in Canada is consistent with an increase in the number of patients treated, an increase in the severity of disease treated, or both. PMID- 8409112 TI - The effect of an oral leukotriene antagonist, ONO-1078, on allergen-induced immediate bronchoconstriction in asthmatic subjects. AB - BACKGROUND: Cysteinyl leukotrienes are potent contractile agonists for airway smooth muscle. To examine the relative role of cysteinyl leukotrienes in immediate airway obstruction in human subjects with asthma after allergen challenge, we investigated the effect of ONO-1078, a potent receptor antagonist of cysteinyl leukotrienes, on the response. METHODS: ONO-1078 and an inactive placebo, in 150-mg capsule form, were administered every 12 hours for 1 week, in a double-blind, cross-over fashion. To examine the effect on the early part of the response, total respiratory resistance (Rrs) was continuously evaluated for 10 minutes after the start of a 1-minute exposure to an appropriate concentration of allergen. Rrs and FEV1.0 were estimated every 10 minutes for 60 minutes after the exposure. RESULTS: ONO-1078 significantly reduced percent changes in Rrs and FEV1.0 from 20 to 60 minutes after the exposure, as well as percent maximum changes in these indices, although the treatment did not alter the time from the start of allergen exposure to the beginning of elevation of Rrs, the time from the start to the point at which total respiratory conductance decreased by 35% from its baseline value, or percent changes in Rrs or FEV1.0 10 minutes after the exposure. CONCLUSIONS: Therefore, we conclude that cysteinyl leukotrienes primarily mediate a later part of immediate airway obstruction after allergen exposure. PMID- 8409113 TI - Aspartame is no more likely than placebo to cause urticaria/angioedema: results of a multicenter, randomized, double-blind, placebo-controlled, crossover study. AB - BACKGROUND: Anecdotes and single case reports have suggested that the high intensity sweetener, aspartame, may be associated with allergic/hypersensitivity type reactions. METHODS: We conducted a multicenter, placebo-controlled clinical study to evaluate individuals who had experienced urticaria and/or angioedema allegedly associated with ingestion of an aspartame-containing product. Despite extensive recruiting efforts over 4 years, only 21 subjects could be enrolled. After admission to clinical research units, subjects were given aspartame and placebo in a randomized, double-blind, crossover fashion. Subjects received, on different days, increasing doses (50, 300, 600 mg) of aspartame and placebo at 8:00 AM, 10:00 AM, and noon. Subjects who weighed less than 40 kg received one half of these doses. Conversion products of aspartame, aspartyl-phenylalanine diketopiperazine and beta-aspartame, were also included in the aspartame arm of the study. Positive reactions were defined as urticaria (hives with wheals 4 mm or more in diameter with a collective diameter of at least 15 mm or one or more hives with a wheal of 4 mm or greater with a flare of 8 mm or greater) or as angioedema. RESULTS: According to these criteria, four reactions were observed; two followed aspartame ingestion and two followed placebo ingestion (p = 1.00). The incidence of other adverse experiences was no different after aspartame versus placebo ingestion (p = 0.289). CONCLUSION: These results indicate that aspartame and its conversion products are no more likely than placebo to cause urticaria and/or angioedema reactions in subjects with a history consistent with hypersensitivity to aspartame. PMID- 8409114 TI - Eosinophilic and neutrophilic inflammation in asthma, chronic bronchitis, and chronic obstructive pulmonary disease. AB - BACKGROUND: Eosinophils but not neutrophils may play a role in the airway inflammation of asthma. In chronic bronchitis (CB) and chronic obstructive pulmonary disease (COPD), neutrophils are present in the airways. To differentiate among the pathology of asthma, CB, and COPD eosinophils and neutrophils were studied in peripheral blood, bronchial biopsy specimens, and bronchoalveolar lavage fluid (BALF). METHODS: We studied nine nonsmoking healthy subjects, 20 nonsmoking patients with asthma, 10 nonatopic smoking patients with CB (forced expiratory volume in 1 second: 98.4% +/- 11.3%) and 17 patients with COPD (forced expiratory volume in 1 second: 51.2% +/- 14.3%). Eosinophils were characterized by their enumeration in biopsy specimens (EG2 monoclonal antibody), peripheral blood, and BALF and by measurement of eosinophil cationic protein in BALF. Neutrophils were characterized by their enumeration in biopsy specimens (anti-elastase monoclonal antibody) and BALF and by measurement of neutrophil specific myeloperoxidase in BALF. RESULTS: In patients with asthma we found degranulated eosinophils in biopsy specimens and significantly increased eosinophil cationic protein levels in BALF. In patients with CB or COPD, eosinophil numbers in biopsy specimens were not significantly different from those of patients with asthma, but cells were not degranulated and eosinophil cationic protein levels in BALF were similar to those of normal subjects. In patients with CB or COPD neutrophils were not increased in the mucosa, but neutrophil numbers and myeloperoxidase levels in BALF were significantly increased. CONCLUSION: The percentages of neutrophils in BALF were greater in patients with COPD than in those with CB, suggesting a role in the chronic airflow limitation. PMID- 8409115 TI - Use of monoclonal antibodies to isolate and characterize Cyn d I, the major allergen of Bermuda grass pollen. AB - BACKGROUND: Cyn d I has been found to be the major allergen of Bermuda grass (Cynodon dactylon) pollen, but its exact nature remains to be clarified. METHODS: Cyn d I, the major allergen of Bermuda grass (Cynodon dactylon) pollen, was purified by monoclonal antibody (MoAb) affinity chromatography, and its biochemical and immunologic properties were characterized. Anti-Cyn d I MoAb 4 37, which recognizes all of the isoallergens of Cyn d I, was chosen as the immunosorbent. RESULTS: The purified protein has an amino acid composition similar to that of the group I allergens of other grass pollens. It appears as a single 34 kd band or as a mixture of 34 and 29 kd polypeptides in sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. The hydrophobicity of these two polypeptides is similar because they have the same retention time on a C18 reverse-phase column when a trifluoroacetic acid/H2O/CH3CN buffer system is used. The N-terminal amino acid sequence of the 34 kd component has a 60% homology with residues of 1-25 of Lol p I, whereas that of the 29 kd component has a 68% homology with residues 31-68 of Lol p I. In addition, this 29 kd polypeptide can be recognized by another anti-Cyn d I MoAb 1-61. CONCLUSIONS: These results suggest that the 29 kd component is derived from Cyn d I. In spite of the similarity in the amino acid composition between Cyn d I and group I allergens of other grass pollens, none of our four anti-Cyn d I MoAbs cross reacted with 10 other grass pollens tested, including ryegrass pollen. Despite biochemical similarity with other group I allergens, the B-cell epitopes on Cyn d I are different from those on other grass pollens. PMID- 8409116 TI - Desensitization of the adenylyl cyclase system in peripheral blood mononuclear cells from patients with asthma three hours after allergen challenge. AB - BACKGROUND: Bronchial hyperreactivity is a common characteristic of patients with asthma and is often associated with enhanced activities of peripheral blood cells. Signal transduction systems are important in regulating cellular activities and can be modified by allergen challenge. METHODS: Patients with allergic asthma (n = 15) were challenged with house dust mite allergen, resulting in an asthmatic response. Adenylyl cyclase activity was measured in membranes from peripheral blood mononuclear cells before, 3 hours after, and 24 hours after challenge. RESULTS: Allergen challenge proved to have opposite effects in two distinct subgroups of patients. In 10 patients (group I) a heterologous desensitization of the adenylyl cyclase system was observed after challenge, whereas in five patients (group II) an increase in adenylyl cyclase activity was found. Adenylyl cyclase activity before allergen challenge in group II was significantly lower than in group I and comparable to cyclase activity found in group I after allergen challenge. This suggests that in these five patients the adenylyl cyclase system was already desensitized before the start of the study, possibly as a result of natural allergen exposure. Heterologous desensitization in group I was found within 3 hours after allergen challenge, that is before the onset of the late bronchoconstrictive reaction. CONCLUSIONS: Because adenylyl cyclase is important in the regulation of cytokine production by mononuclear cells, alteration of cytokine production induced by desensitization of adenylyl cyclase could therefore play a role in the development of the late bronchoconstrictive reaction. PMID- 8409117 TI - The use of murine polyclonal anti-idiotypic antibodies as surrogate allergens in the diagnosis of Parthenium hypersensitivity. AB - BACKGROUND: Anti-idiotypic antibodies (Ab-2), which are the mirror images of idiotypic antibodies (Ab-1), may be useful as diagnostic reagents and for use as immunogen to induce antigen-specific immune responses. METHODS AND RESULTS: To explore the biologic potential of Ab-2 as diagnostic reagents in allergic diseases, murine mouse (m) Ab-2 were raised by immunizing Balb/c mice with affinity purified rabbit (r) Ab-1 specific for the pollen of Parthenium hysterophorus, an allergenic weed that grows wild on the Indian subcontinent and in Australia, Mexico, and the southern United States. Affinity purified Parthenium-specific human (h)AB-1 could successfully inhibit the binding of mAb-2 to immobilized rAb-1. Further, Balb/c mice immunized with mAb-2 induced Parthenium-specific anti-anti-idiotypic IgE and IgG antibodies. Specificity of the Ab-2 was confirmed by the ability of Parthenium pollen extracts to inhibit the binding of allergen-specific IgE and IgG Ab-1 in the sera of patients with rhinitis to immobilized mAb-2. Parthenium-sensitive patients with rhinitis who had positive results on skin prick tests to Parthenium pollen extracts also responded with a positive skin reaction to mAb-2. CONCLUSION: Our data demonstrate that Parthenium-specific mAb-2 may be of value as surrogate allergens in allergen standardization and for in vitro diagnosis. PMID- 8409118 TI - In vivo detection of a novel macrophage-derived protein involved in the regulation of nasal mucus-like glycoconjugate secretion. AB - BACKGROUND: We recently described a novel 68 kd mucus secretagogue (MMS-68) derived from human monocytes, pulmonary macrophages, and a macrophage hybridoma, clone 63. We detected MMS-68 in monocyte culture supernatants from patients with steroid-dependent asthma and in bronchoalveolar lavage fluid from patients with chronic bronchitis by antigen capture ELISA and in normal lung tissue by immunohistochemistry. METHODS: To determine a role for MMS-68 in the regulation of nasal mucus, we labeled human nasal turbinates with tritiated glucosamine and assayed for the ability of the previously purified MMS-68 (stock solution) to induce mucus-like glycoconjugate release (MLGC). We also performed immunohistochemistry stains with an anti-MMS-68 antibody (1-D-10) on frozen sections (n = 5) of nasal turbinates from patients with allergic and nonallergic rhinitis who were undergoing rhinoplasty and measured MMS-68 levels in nasal lavages from patients who were undergoing topical nasal histamine or methacholine challenge. RESULTS: MMS-68 is a potent nasal MLGC secretagogue causing a dose dependent increase in MLGC release in vitro. Staining revealed a subepithelial distribution for MMS-68. Antigen capture ELISA of nasal lavages demonstrated mean MMS-68 levels from saline control challenge of 0.9 +/- 0.5 micrograms MMS-68 per milligram of protein (n = 5), 8.6 +/- 1.4 micrograms MMS-68 per milligram of protein from histamine challenge and 20.7 +/- 2.3 micrograms MMS-68 per milligram of protein (n = 5) after methacholine challenge. CONCLUSION: Taken together these data suggest that MMS-68 may play a role in the normal regulation of mucus secretion. PMID- 8409119 TI - Eosinophil major basic protein enhances the expression of neutrophil CR3 and p150,95. AB - BACKGROUND: We examined the effect of major basic protein (MBP) stimulation on the expression of the neutrophil B2-integrins: LFA-1 (CD11a/CD18), CR3 (CD11b/CD18), and p150,95 (CD11c/CD18). METHODS: Incubation of neutrophils with 0.75 to 3.0 mumol/L MBP for 30 minutes at 37 degrees C resulted in concentration dependent increases in CR3 expression as detected by staining with the CD11b specific monoclonal antibody, Leu-15. RESULTS: The expression of CR3 was significantly (p < 0.001) higher when neutrophils were stimulated with 1.5 mumol/L (53% +/- 9% increase) or 3.0 mumol/L (100% +/- 17% increase) MBP as compared with unstimulated neutrophils. The kinetics of MBP-stimulated CR3 expression were rapid and were similar to those of 100 nmol/L. N-formyl-methionyl leucyl-phenylalanine-stimulated enhancement of CR3 expression. Incubation of neutrophils with reduced and alkylated MBP resulted in significantly lower (p < 0.05) increases in CR3 expression as compared with stimulation with native MBP. In addition, neither eosinophil cationic protein nor eosinophil-derived neurotoxin altered neutrophil CR3 expression. MBP stimulated minimal increases in LFA-1 expression. However, staining with the monoclonal antibody Leu-M5 (anti CD11c) revealed that MBP stimulated significant increases in the expression of p150,95. CONCLUSIONS: These results indicate that MBP stimulates the increased expression of neutrophil adhesion molecules, which in turn may enhance the inflammatory role of neutrophils in the late-phase events of allergic diseases. PMID- 8409120 TI - Eosinophil survival activity identified as interleukin-5 is associated with eosinophil recruitment and degranulation and lung injury twenty-four hours after segmental antigen lung challenge. AB - BACKGROUND: Segmental antigen challenge in allergic volunteer subjects leads to the recruitment of inflammatory cells, including eosinophils, to the lung and to lung injury as shown by albumin influx into the alveolar air space. The goal of this study was to determine whether eosinophil-active cytokines, including IL-3, IL-5, or granulocyte-macrophage colony-stimulating factor, are released into the lung 24 hours after segmental antigen challenge of ragweed allergic subjects with allergic rhinitis and to determine whether the presence of the cytokine or cytokines is correlated with markers of lung inflammation and lung injury. METHODS: Volunteers underwent challenge with a wide variety of antigen doses, which resulted in the recruitment of inflammatory cell mixtures both with and without eosinophils. RESULTS: Eosinophil survival activity (ESA), the ability of the cytokine to prolong blood eosinophil survival in culture, was found in 5 of 17 ragweed allergic subjects and only in subjects challenged with relatively high doses of ragweed antigen (0.2 ragweed antigen units/ml or more). No ESA was found in bronchoalveolar lavage (BAL) fluid in any of eight nonragweed allergic subjects. This activity could be almost completely neutralized by preincubating BAL fluid with specific antibody to IL-5, although a small contribution by granulocyte-macrophage colony-stimulating factor may also have been present. ESA correlated with eosinophil recruitment (r = 0.72, p < 0.001) and degranulation in the lung (r = 0.63 to 0.81, p < 0.01, for eosinophil granule constituents in BAL fluid) and lung injury as shown by albumin influx into the alveolar air spaces (r = 0.83, p < 0.001). ESA was unrelated to the presence of other inflammatory cells in the lung. Subjects who had IL-5 in BAL fluid appeared to undergo more severe initial reactions to antigen challenge. CONCLUSIONS: We conclude that IL-5 is the most important constituent in ESA in the lung 24 hours after antigen challenge and that it correlates with eosinophil recruitment, degranulation, and lung injury. PMID- 8409121 TI - Effects of environment on atopic status and respiratory disorders in children. AB - BACKGROUND: We investigated whether living in areas with higher air pollution levels increases the prevalence of positive skin reactivity in children and the possible synergic effect of air pollution exposure and atopy on respiratory disorders. METHODS: This cross-sectional study was conducted in an urban area, in an industrialized area, and in a rural control area in the Latium region of Italy. A total of 2226 subjects, aged 7 to 11 years, were studied. RESULTS: The prevalence of children with positive skin test results did not vary significantly over the areas (urban area = 21.0%, industrialized area = 22.0%, rural area = 20.2%). Children living in polluted areas experienced significantly more cough and phlegm (odds ratio [OR] = 1.5), rhinitis (OR = 1.7), pneumonia (OR = 1.7), and early respiratory infections (OR = 1.4) than control subjects. The pattern of the odds ratios for atopy and air pollution suggested that the two factors were probably additive in affecting asthma and early respiratory infections (synergy index [SI] = 1.04 and 1.27, respectively), whereas they seemed to act synergically in regard to cough and phlegm (SI = 1.59), rhinitis (SI = 3.01), and pneumonia (SI = 2.75). CONCLUSION: Environmental air pollution seems not to increase the prevalence of atopic status; it seems, however, to enhance the development of clinical symptoms among already sensitized subjects. PMID- 8409122 TI - Fusarium allergic fungal sinusitis. PMID- 8409123 TI - Bird-egg syndrome in childhood. PMID- 8409124 TI - Socioeconomic status and allergy in children with asthma. PMID- 8409125 TI - Putting a dietetics spin on health care issues. PMID- 8409126 TI - Differences on treating children with galactosemia. PMID- 8409127 TI - RDs and restaurants: recipe for success. How to sell consulting services to restaurants. PMID- 8409128 TI - Finger foods help those with Alzheimer's maintain weight. PMID- 8409129 TI - Labeling of dietary supplements stirs Congressional debate. PMID- 8409130 TI - Managed health care: understanding the role of the nutrition professional. PMID- 8409131 TI - Standards of practice for the nutrition support dietitian: importance and value to practitioners. AB - OBJECTIVE: To determine whether the American Society for Parenteral and Enteral Nutrition (ASPEN) standards of practice for nutrition support dietitians reflect current practice. DESIGN: A mailed survey questionnaire was used to determine how important nine functions of the nutrition support dietitian are to the practice of nutrition support dietetics, how important the ASPEN standards are to practitioners, the frequency of application of each standard to patients, and demographic data of the respondents. SAMPLE: The survey was sent to 1,048 randomly selected dietitian members of The American Dietetic Association's Dietitians in Nutrition Support dietetic practice group and ASPEN. The final sample included 460 dietitians, a return rate of 44%. For purposes of comparison, dietitian respondents were categorized into three groups: nutrition support dietitians (n = 286); non-nutrition support dietitians (n = 136); and supervisors of nutrition support dietitians (n = 38). STATISTICAL ANALYSES PERFORMED: Descriptive data are reported as frequency or mean +/- standard deviation. Differences in importance ratings of the standards by nutrition support dietitians, non-nutrition support dietitians, and supervisors of nutrition support dietitians were compared with one-way analysis of variance ANOVA and Scheffe post hoc tests. chi 2 Analysis was used to compare group differences in the frequency of application of each standard to patients. Probability was set at the .05 level. RESULTS: Nutrition support functions rated as very important included performing nutrition assessments, identifying high-risk patients, implementing and monitoring parenteral and enteral nutrition, supervising transitional feeding, and documentation. The standards of practice were all rated as very important by nutrition support dietitians, non-nutrition support dietitians, and supervisors, and there was no difference in perceived importance. More than half of the respondents applied each standard with 75% to 100% of their patients; however, there was a higher frequency of application by nutrition support dietitians. APPLICATIONS/CONCLUSIONS: The findings give validity to the standards of practice. Practitioners evaluate them as being important in practice and in theory. The standards of practice for nutrition support dietitians were approved for use by the House of Delegates of The American Dietetic Association in October 1992. PMID- 8409132 TI - Use of eating-pattern messages to evaluate changes in eating behaviors in a worksite cholesterol education program. AB - OBJECTIVE: To determine whether eating-pattern messages can effectively be used in a worksite cholesterol education program to change eating behaviors. SUBJECTS: 91 randomly selected participants with initial serum cholesterol levels of 5.2 mmol/L attended the program. INTERVENTION: Eating-pattern messages were the focus of a successful 8-week worksite cholesterol education program conducted with city employees of Phoenix, Ariz. Participants completed self-administered questionnaires before and after the intervention that asked them to compare their current eating patterns with those addressed in the program. The majority (n = 84) of the participants attended five or more of eight available sessions, led by registered dietitians, which focused on the skills needed to decrease dietary fat. STATISTICAL ANALYSES PERFORMED: Parametric and nonparametric statistical tests were used to evaluate the direction and magnitude of changes in eating patterns. RESULTS: Participants made statistically significant changes in 11 of 15 eating patterns linked to messages delivered during the intervention. Changes in eating behaviors were related to improvements in blood lipid profiles. Results from a multiple regression analysis indicated that intervention-related changes in total cholesterol were significantly associated with combined eating-pattern message scores, and total cholesterol decreased 0.33 mmol/L for each unit decrease in the combined eating-pattern message score. APPLICATIONS/CONCLUSIONS: These findings indicate that eating-pattern messages can be used successfully to evaluate changes in fat-related eating behaviors. PMID- 8409133 TI - Comparison of the effects of adding fish high or low in n-3 fatty acids to a diet conforming to the Dietary Guidelines for Americans. AB - OBJECTIVE: To compare the effects of diets containing high or low amounts of n-3 fatty acids on lipid levels in men. DESIGN: A continuous 40-day feeding study was conducted using a 5-day cycle menu. The control diet, which was fed the first 21 days of the trial, contained 32% of energy as fat and 44.4 g total dietary fiber per day. Total serum cholesterol (TC), low-density lipoprotein cholesterol (LDL C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TGs) were evaluated 1 day before the trial began and on days 21 and 41. SUBJECTS: 17 men aged 21 to 42 years who were free-living individuals with no apparent illnesses or hyperlipidemia. INTERVENTION: The treatment diets, which were substituted daily for the regular entrees for the last 19 days of the trial, contained 180 g/day of either farm-raised catfish or wild Alaskan salmon. STATISTICAL ANALYSES PERFORMED: Serum data were analyzed using analysis of covariance with the initial values as the covariates. RESULTS: After 21 days on the control diet, TC and TGs decreased; however, there were no further decreases during the last 19 days when fish was consumed. LDL-C remained unchanged after the first 21 days but was lower than the initial value by day 41. HDL-C levels were similar over the 40 days. There were no significant differences in lipid levels of men who consumed catfish and those who consumed salmon. APPLICATIONS/CONCLUSIONS: Farm-raised catfish compared favorably to wild Alaskan salmon when incorporated into the total diet in regard to lipid levels in men. PMID- 8409134 TI - Food preferences and food habits of patients with chronic renal failure undergoing dialysis. AB - OBJECTIVE: To examine and compare the food preferences of patients undergoing maintenance hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) with those of age- and sex-matched controls. DESIGN: We administered two questionnaires, one assessing preferences for 88 food items (according to a nine point hedonic scale) and the other assessing factors influencing dietary habits. SUBJECTS: Thirty-three patients on HD, 17 patients on CAPD, and 30 control subjects with normal renal function. MAIN OUTCOME MEASURES: The 88 foods were grouped into 14 classes to compare preferences. Taste aversions, food preparation details, and psychological and social determinants of food intake were also compared. STATISTICAL ANALYSIS: The preference ratings of the HD, CAPD, and control groups were compared using analysis of variance. The dietary habits results were examined using a chi 2 analysis. RESULTS: Sweet foods (P = .002), vegetables (P = .003), red meats (P = .010), and fish and poultry (P = .015) were less pleasant for patients on HD than for control subjects. Red meats (P = .019), fish and poultry (P = .032), and eggs (P = .005) were less pleasant for patients on HD than for patients on CAPD. Red meat was the most unpopular food group for all dialysis patients. The most common factor affecting dietary intake was a loss of interest in food and/or cooking. CONCLUSION: We conclude that chronic renal failure influences patients' food preferences and food habits. Knowledge of these preferences will help dietitians plan more adequate and enjoyable diets for such patients. PMID- 8409135 TI - Height, weight, and body mass index of American Indian schoolchildren, 1990-1991. AB - OBJECTIVE: To describe the current height and weight status of American Indian children who live on or near Indian reservations nationwide. SUBJECTS: 9,464 American Indian schoolchildren aged 5 through 18 years. STATISTICAL ANALYSES PERFORMED: Data for height, weight, and body mass index of the schoolchildren were compared with two national reference data sets, the second National Health and Nutrition Examination Survey (NHANES II) and the Mexican-American population of the Hispanic Health and Nutrition Examination Survey (HHANES-MA). RESULTS: The three populations were similar in height, but the American Indian children weighed more, although not at a statistically significant level, and had a statistically significant higher body mass index than the NHANES II reference population for nearly every age and sex group. The overall prevalence of overweight in the American Indian children (exceeding the 85th percentile of the reference population) was 39.3% compared with the NHANES II population and 28.6% compared with the HHANES-MA population. The overall prevalence of underweight in the American Indian sample was substantially less than the expected 15% compared with either the NHANES II or HHANES-MA population. APPLICATIONS/CONCLUSIONS: Overweight is much more prevalent in American Indian children than among other children in the United States at all ages and in both sexes. This may have important implications for chronic disease risk and emphasizes the need for targeting obesity prevention efforts to American Indian children. PMID- 8409136 TI - Job sharing in clinical nutrition management: a plan for successful implementation. AB - While women continue to enter the American work force in record numbers; many experience difficulty in juggling career and family obligations. Flexible scheduling is one option used to ease work and family pressures. Women's changing work roles have potentially noteworthy implications for clinical nutrition management, a traditionally female-dominated profession where the recruitment and retention of valued, experienced registered dietitians can prove to be a human resources challenge. Job sharing, one type of flexible scheduling, is applicable to the nutrition management arena. This article describes and offers a plan for overcoming obstacles to job sharing, including determining feasibility, gaining support of top management, establishing program design, announcing the job share program, and using implementation, monitoring, and fine-tuning strategies. Benefits that can be derived from a successful job share are reduced absenteeism, decreased turnover, enhanced recruitment, improved morale, increased productivity, improved job coverage, and enhanced skills and knowledge base. A case study illustrates one method for achieving job sharing success in clinical nutrition management. PMID- 8409137 TI - Home health care: a new worksite for dietitians monitoring nutrition support. AB - Dietitians have been practicing in the home setting for many years. However, monitoring patients receiving home parenteral and enteral nutrition has been performed primarily on an outpatient basis by dietitians affiliated with hospital based nutrition support teams. Changes in physician familiarity with these specialized therapies and expansion of the home infusion therapy industry have resulted in opportunities for dietitians to monitor nutrition support in a patient's home. This article describes the role of the home nutrition support dietitian, the work environment, and the training needed to prepare the practitioner for effective work in this field. Practical concerns of interest to the dietitian monitoring home nutrition support include equipment, resources, and communication tools. Home visits impact several benefits to dietetics practice by enriching the contact between patient and dietitian. A case study describes a dietitian's involvement in and potential cost-effectiveness of treatment of a patient whose parenteral nutrition therapy was initiated and completed without hospitalization. The home is emerging as a worksite for dietitians who monitor nutrition support. As providers of home infusion therapy continue to expand, widespread availability of dietitians' services for patients receiving parenteral and enteral support at home must be ensured. PMID- 8409138 TI - Preschoolers' reporting of food habits. PMID- 8409139 TI - Operational characteristics of hospital foodservice departments with conventional, cook-chill, and cook-freeze systems. PMID- 8409140 TI - Dexfenfluramine influences dietary compliance and eating behavior, but dietary instruction may overrule its effect on food selection in obese subjects. PMID- 8409141 TI - Longitudinal changes in zinc status in untrained men: effects of two different 12 week exercise training programs and zinc supplementation. PMID- 8409142 TI - Undocumented potential drug interactions found in medical records of elderly patients in a long-term-care facility. PMID- 8409143 TI - Position of The American Dietetic Association: nutrition services in health maintenance organizations and other forms of managed care. PMID- 8409144 TI - Debra A. Krummel wins Huddleson Award. PMID- 8409145 TI - A randomized, controlled trial of outpatient geriatric evaluation and management in a large public hospital. AB - OBJECTIVE: To study the effect of outpatient geriatric evaluation and management on physical function, mental status, and subjective well-being. DESIGN: Prospective randomized controlled trial with a 1-year study period. SETTING: Large medical school-affiliated public hospital in an urban community. SUBJECTS: Patients at least 70 years old admitted to the medicine service were screened, randomized, and completed a 1-year follow-up interview. INTERVENTIONS: Comprehensive geriatric evaluation and an outpatient care management program. MAIN OUTCOME MEASURES: Mental status (SPMSQ), ADL (Katz Index), IADL (Five-Item OARS Scale), Life satisfaction (LSI-Z), and self-perception of health status (physical health section of OARS). RESULTS: No significant differences were found for cognitive status, ADL functioning, life satisfaction, nursing home placement, or mortality. The experimental patients reported significantly higher function in IADL and more favorable self-perception of health status compared with controls. CONCLUSION: Outpatient comprehensive geriatric evaluation and management appears to be a useful model for providing care to medically frail elderly patients. PMID- 8409146 TI - A longitudinal study of cognitive function in elderly persons with subjective memory complaints. AB - OBJECTIVE: To assess change in cognitive function in elderly individuals with subjective memory loss over a follow-up interval of more than 3 years. DESIGN: Prospective cohort study. SETTING: An outpatient research clinic in aging and dementia at a university hospital. PARTICIPANTS: A convenience sample of 59 healthy, elderly individuals (mean age 68.7 years) with memory complaints but no clinically apparent cognitive dysfunction. MEASUREMENTS: Participants were given a full diagnostic evaluation and were administered a neuropsychological test battery at baseline and follow-up. The cognitive assessment battery included 12 tests of recent memory, immediate memory, language function, visuospatial praxis, and psychomotor speed. Most of the tests had been established to be sensitive to cross-sectional age differences. RESULTS: Of 59 subjects, 54 (91.5%) were successfully followed up an average of 3.4 years later. Between baseline and follow-up, two tests exhibited significant improvement, two tests exhibited significant decline, and the other eight were unaffected. CONCLUSION: Elderly individuals with subjective perceptions of cognitive decrements who fail to provide clear evidence of cognitive impairment upon clinical interview are not at high risk for progressive cognitive deterioration over the subsequent 3- to 4 year interval. PMID- 8409147 TI - Projecting the need for physicians to care for older persons: effects of changes in demography, utilization patterns, and physician productivity. AB - OBJECTIVE: To determine the influence of differing assumptions of population growth, visit rates, prevalence of functional impairment, physician productivity, and hospitalization rates on projected need for physicians to provide medical care for older persons. DESIGN: Sensitivity analysis of a manpower model. MAIN RESULTS: The factors that appear to have the most impact on projections of physician need are related to physician productivity, especially delegation to mid-level providers, and case-mix. Other factors, such as the variability of census projections and per capita visit rates, are likely to have less effect on overall physician supply needs. CONCLUSIONS: Although case mix and delegation to mid-level providers may both substantially affect the need for physician supply to care for older persons, only the latter can be directly affected by health policy decisions. Consideration should be given to increasing the supply of mid level providers and providing incentives for patients and physicians to receive and provide care in delivery systems that utilize mid-level providers extensively. PMID- 8409148 TI - Predictors of family caregivers' physical and psychological health following hospitalization of their elders. AB - OBJECTIVE: To identify characteristics present at the time of elders' (> or = 75 years of age) hospitalization that predicted care-givers' self-reported physical and psychological health 1 year later. DESIGN: A prospective observational cohort study of 104 family caregivers followed for 1 year from the time of their elders' hospitalization. SETTING: Urban community hospital with academic affiliation. SUBJECTS: Caregivers of patients enrolled in an evaluation of inpatient interdisciplinary geriatric assessment who provided care for the entire year of follow-up and who participated in all interviews. MAIN OUTCOMES: Self-reported physical and psychological health. RESULTS: Caregivers' baseline self-reported physical health (AOR = 21.8; 95% CI = 4.65-102), the impact of caregiving on their social and leisure time activities (AOR = 9.93; 95% CI = 1.71-57.8), and their relationship to their elders (AOR = 6.50; 95% CI = 1.20-35.2) were all significantly associated with self-reported physical health at follow-up. Caregivers' baseline self-reported health (AOR = 3.70; 95% CI = 1.11-12.5), whether or not they had additional current caregiving responsibilities (AOR = 6.67; 95% CI = 1.89-25.0), and the interaction of perceptions of caregiving and the adequacy of family help were significant predictors of psychological health at follow-up, adjusted for baseline psychological health. CONCLUSION: Information available at the time of elderly patients' admission to the hospital can help to differentiate those caregivers at risk for future physical and psychological distress from those who are likely to continue doing well. PMID- 8409149 TI - Age-related trends in cardiovascular morbidity and physical functioning in the elderly: the Cardiovascular Health Study. AB - OBJECTIVE: To describe relationships between age and sub-clinical cardiovascular disease, manifest chronic disease, and physical functioning and limitations among persons aged 65 years and older, with emphasis on the "oldest old," those 85 years and older. DESIGN: Observational population-based study. SETTING: Four U.S. communities: Forsyth County, North Carolina; Sacramento County, California; Washington County, Maryland; and Pittsburgh, Pennsylvania. PARTICIPANTS: 5,201 men and women aged 65 years and older. MEASUREMENTS: Demographic data; histories of cardiovascular disease (CVD), chronic lung disease, arthritis, diabetes, and hypertension; measures of subclinical disease including arm and ankle blood pressures, internal carotid wall thickness and stenosis, ejection fraction, left ventricular mass, fractional shortening, and diastolic function, electrocardiographic left ventricular hypertrophy and cardiac injury score, forced expiratory flow and volume; functional status including self-reported physical functioning, hearing and sight limitations and health status, and performance-based measures of function. These variables were examined among men and women in three age groups: 65-74 years, 75-84 years, and 85 + years. Subgroups of participants with and without manifest CVD were also examined. MAIN RESULTS: In women, the prevalence of CVD and other chronic conditions increased with age, and the highest rates occurred among those 85 years and older. In men, prevalence rates increased between the two younger groups, but the oldest group had lower than expected rates for coronary heart disease, cerebrovascular disease, hypertension, and chronic lung disease. In contrast, there were strong age-related linear trends in most of the subclinical measures of blood pressure, atherosclerosis and pulmonary function and in virtually all measures of functional status in both gender groups across the age range. There was a particularly marked decline in functional status between the two older age groups. While subclinical disease was greater and functional status was poorer among those with manifest CVD, with few exceptions, age-related trends were not significantly different between the two groups. CONCLUSIONS: Lower than expected prevalence rates of CVD among those aged 85 years and older, particularly among men, in this study of community-dwelling elderly may represent selection bias or a real plateauing in disease prevalence with age. However, subclinical disease appears to increase and functional status to decline across the age range in both men and women regardless of the presence of CVD. The apparent increase in subclinical disease with age indicates potential for CVD prevention after age 65. PMID- 8409150 TI - Calcium homeostasis of an elderly population upon admission to a nursing home. AB - OBJECTIVE: To evaluate the skeletal health, calcium, and vitamin D homeostasis of patients upon their entry to a long-term-care facility. Factors that could contribute to the risk of future osteoporotic fractures were also evaluated. DESIGN: Cross-sectional study. PATIENTS: Two hundred eighty-seven consecutive admissions were invited to participate; 109 patients were recruited into the study. MEASUREMENTS: A high prevalence of low to low-normal circulating levels of 25(OH)D was found in nursing home residents upon their admission to the nursing home, with 86% of the patients having circulating 25(OH)D levels of less than 50 nmol/L and 41% having levels below 25 nmol/L. Frankly elevated parathyroid hormone levels were found in 16% of the patients. Additionally, alkaline phosphatase and osteocalcin levels were elevated in 23% and 13% of the patients, respectively. Bone mineral measurements were in the osteoporotic range for 85% of the nursing home residents. Bone density results for females with a history of any classic osteoporotic fracture were significantly lower than for those with no fracture history (68.5 arbitrary units (AU) for those with no fracture history, 58.8 AU in those with history of hip fracture; P < 0.05). The bone mineral measurements were higher in women with adequate 25(OH)D compared with women with deficient or borderline 25(OH)D with and without fracture history. Levels of 25(OH)D were positively correlated with urine calcium/creatinine (r = .24; P = 0.03) and 1,25(OH)2D (r = .28; P = 0.01) and were negatively correlated with 1-84 PTH (r = -.24; P = 0.02). CONCLUSION: Hypovitaminosis D is prevalent among elderly patients entering a nursing home with secondary hyperparathyroidism and apparently increased bone turnover present in patients with circulating 25(OH)D levels below 50 nmol/L. Bone density measurements showed that a majority of the individuals entering a nursing home are osteoporotic. There is a positive association between 25(OH)D levels and bone mass and a negative association between 25(OH)D levels and a history of fracture. PMID- 8409151 TI - Cognitive impairment and the use of health services in an elderly rural population: the MoVIES project. Monongahela Valley Independent Elders Survey. AB - OBJECTIVE: To describe the patterns of use of health and human services by elderly rural individuals and to determine whether cognitively impaired persons had a distinctive pattern of service utilization. DESIGN: An epidemiological survey and cognitive screening of an age-stratified random community sample. SETTING: The mid-Monongahela Valley, a rural community in Southwestern Pennsylvania. PARTICIPANTS: 1366 non-institutionalized persons aged 65 years and older, fluent in English, and with at least a sixth grade education. MEASUREMENTS: We administered a battery of cognitive screening tests (the Mini Mental State Examination; Immediate and Delayed Recall of a Story; Immediate and Delayed Recall of a Word List; CERAD modification of the Boston Naming Test; Verbal Fluency for initial letters P and S and for names of Fruits and Animals; Temporal Orientation; Praxis; Clock Drawing; and Trailmaking Tests A and B.) We also obtained basic demographic information and inquired about the use of health and human services in the previous year. RESULTS: Approximately 10% of the sample was classified (by operational criteria) as cognitively impaired. In univariate analyses, cognitive impairment was found to be significantly associated with the use of certain health services: hospitalization in the previous 6 months (odds ratio, OR = 2.1; 95% CI = 1.3, 3.3), previous nursing home use (OR = 9.3; 95% CI = 3.8, 22.9), home health care (OR = 4.6; 95% CI = 2.7, 8.0), social services (OR = 6.5; 95% CI = 3.4, 12.4), mental health services (OR = 2.8; 95% CI = 1.2, 6.2), and the regular use of prescription medications (OR = 2.0; 95% CI = 1.3, 3.2). Visits to physicians were not significantly different between the impaired and unimpaired groups. In a multiple regression model, which included age and educational level, the use of home health care (OR = 3.4; 95% CI = 1.8, 6.4) and social services (OR = 2.3; 95% CI = 1.1, 4.9) remained significantly associated with cognitive impairment. CONCLUSIONS: The association of cognitive impairment with the use of these health and human services underscores the general frailty of the impaired group. These findings also point to potential target groups in the community for further evaluation and services for dementia. The findings support the need for education regarding cognitive impairment and dementia to be made available to providers of these services, particularly informal social services such as those provided by church groups. PMID- 8409152 TI - The use of intramuscular cefoperazone versus intramuscular ceftriaxone in patients with nursing home-acquired pneumonia. AB - OBJECTIVE: To compare the efficacy and safety of intramuscular cefoperazone and intramuscular ceftriaxone in the treatment of nursing home-acquired pneumonia in the nursing home setting. DESIGN: A randomized clinical trial. SETTING: Skilled nursing wards at the Veterans Home of California. PATIENTS: 104 residents of skilled nursing wards, aged 65 years or older. INTERVENTIONS: Intramuscular administration of either cefoperazone or ceftriaxone. MEASUREMENTS: The variables analyzed for baseline comparability were demographics (age, sex), clinical variables (duration in nursing home; presence of sputum, fever, cough, or leukocyte count), and clinical symptoms and signs. Efficacy was assessed by days of therapy, final maximum temperature, and clinical and bacteriological response. RESULTS: Fifty residents received cefoperazone, 1 gm every 12 hours, intramuscularly. Fifty-four residents received ceftriaxone, 1 gm every 24 hours, intramuscularly. The total duration of treatment was scheduled for 10 days. Clinical cure was seen in 45 (90%) of the cefoperazone treatment group and 51 (94%) of the ceftriaxone treatment group, with a mean duration of therapy of 10.30 and 9.90 days, respectively. Satisfactory sputum specimens were collected in 71% of the treated residents; the most common isolate was Streptococcus pneumoniae, followed by Haemophilus influenzae and Staphylococcus aureus, respectively. The overall mortality was 4.5% at long-term follow-up. Both agents were well tolerated and no therapy was discontinued due to intramuscular pain or abnormal laboratory values. CONCLUSIONS: Intramuscular cefoperazone and intramuscular ceftriaxone are safe and effective in the treatment of nursing home acquired pneumonia. The clinical outcomes in both treatment groups support their use within this select population without the need for transferring the patient to an acute care hospital. Clinical studies are needed to evaluate the impact of such therapy on the control of health care expenditures within the nursing home facility. PMID- 8409153 TI - Age-related variability in the use of cardiovascular imaging procedures. AB - OBJECTIVE: To determine if older patients undergo fewer cardiovascular imaging procedures (CIPs) than younger patients when admitted to a tertiary care academic medical center for an acute myocardial infarction (MI), after adjusting for disease severity and comorbidities. DESIGN: Non-current prospective cohort study. SETTING: Urban tertiary care academic medical center. PATIENTS: Medical records of 294 patients admitted and diagnosed with an acute MI between January 1990 and April 1991 were reviewed. MEASUREMENTS: The total number of different CIPs performed during hospitalization was determined. Cardiac catheterizations, echocardiograms, radionuclide ventriculograms, and thallium scans counted as CIPs. Disease severity was assessed by the Acute Physiology Score (APS) of APACHE II, admission Killip's Classification, and peak creatine phosphokinase (CPK) levels. Comorbidities were assessed using a modified Comorbidity Damage Index of Charlson. RESULTS: The mean (+/- SD) number of different CIPs performed during hospitalization was significantly less for those > or = 75 years old (1.3 +/- 1.0) than for those < 75 years old (1.7 +/- 1.0) (P = 0.01), and CIP number negatively correlated with age (Spearman r = -0.178; P = 0.01). Mean CIP number decreased from 2.0 +/- 1.1 for those < 45 years old to 0.9 +/- 0.6 for those > or = 85 years old (P = 0.02). Other factors positively associated (P < 0.10) with CIP number were: CPK values in the highest quartile of the study population (> 355 U/L); admission to a cardiology, medical, or family practice service; no CIP performed at an outside hospital prior to transfer; admission Killip's Classification of less than IV, and a Q-wave MI. After adjusting for these variables in a multiple regression model, age > or = 75 remained an independent predictor of decreased CIP use (P = 0.003). The modified comorbidity index score and the APS score, a general measure of severity of illness, were not significantly associated with CIP use. When procedures were examined individually, no significant age-related differences were noted in the use of thallium scans, radionuclide ventriculograms, or echocardiograms. Older patients did, however, remain less likely to undergo cardiac catheterizations (P < 0.001). CONCLUSION: Older patients, regardless of underlying disease severity or comorbidities, undergo fewer invasive cardiovascular evaluations than younger patients when admitted to a tertiary care academic medical center for an acute MI. PMID- 8409154 TI - Urinary incontinence in nursing homes: incidence, remission and associated factors. AB - OBJECTIVE: To determine the incidence and remission rates of daytime urinary incontinence (UI) in a cohort of newly admitted nursing home (NH) residents. DESIGN: Prospective cohort study. SETTING: Eight proprietary NHs in Maryland. SUBJECTS: Four hundred thirty new admissions age 65 or older who were participants in a larger prospective study of mental morbidity and adjustment to the NH. MEASURES: Nurses aides' reports of continence status, psychiatric examinations, and nursing staff assessments of mobility at 2 weeks, 2 months, and 1 year after NH admission. RESULTS: The prevalence of daytime UI at admission was 39% in both females and males. Among the 293 members (68%) of the admission cohort remaining in the NHs 2 months after admission, the incidence of daytime UI was 27% (21% in females, 51% in males); daytime UI resolved in 23% (24% in females, 20% in males). Among the 178 members (41%) of the admission cohort remaining in the NHs 1 year after admission, the incidence of daytime UI between 2 months and 1 year after admission was 19% (16% in females, 46% in males); daytime UI resolved in 22% (23% in females, 14% in males). The continence status of about two-thirds of residents remaining in the NH at 1 year after admission was stable over time: 22% had daytime UI, and 42% were continent at all three data collection points. The development of daytime UI was associated with male sex, the diagnosis of dementia, fecal incontinence, and the inability to ambulate or transfer independently. Resolution of daytime UI was associated with the absence of these characteristics. CONCLUSIONS: Despite limitations attributable to the method of defining UI and potential biases related to the attrition of the admission cohort over time, this is the first large prospective study to examine the incidence and remission patterns of daytime UI among NH residents. The strong association between UI and dementia was validated for the first time by direct psychiatric examinations. Sex and mobility are also closely associated with the development and remission of UI in this setting. This study provides some valuable data that can be used to assess the impacts of the recently developed Resident Assessment Protocol for UI and Agency for Health Care Policy and Research Clinical Practice Guidelines. PMID- 8409155 TI - Serum fructosamine as a screening test for diabetes in the elderly: a pilot study. AB - OBJECTIVE: To determine the value of serum glycated protein, measured as serum fructosamine, as a screening test for diabetes in the elderly. DESIGN: Cross sectional pilot study. SETTING: Ambulatory research clinic in university setting. PATIENTS: One hundred fifty-seven consecutive community-dwelling participants in the Cardiovascular Health Study, average age 71.8 + 5 (mean +/- SD, range 65-88 years). MEASUREMENTS: Serum fructosamine levels (first and second generation assay) were obtained. All subjects who did not have a diagnosis of diabetes were given a 75-g glucose tolerance test (GTT). RESULTS: Twenty-six subjects (17%) (10 previously diagnosed, 16 undiagnosed and asymptomatic) had diabetes mellitus, and 38 subjects (24%) had impaired glucose tolerance by history or by the GTT (WHO criteria). Only the 16 asymptomatic diabetics were included in the analysis for the pilot study. There was a significant difference in the fasting fructosamine level between non-diabetics and asymptomatic diabetics for the first generation (2.06 +/- .21 vs 2.53 +/- .49 mMol/L, P < 0.0015) and second generation assay (221 +/- 27 vs 269 +/- 48 mMol/L, P < 0.0012). Receiver operator curves were constructed to evaluate the test characteristics of serum fructosamine. Using a point of > or = 2.3 mMol/L for the first-generation assay, the sensitivity to detect asymptomatic diabetes was 75%, specificity 83%, and positive predictive value 35%. To detect both diabetes and impaired glucose tolerance using a cutpoint of > or = 2.3 mMol/L, the sensitivity was 24%, specificity 95%, and positive predictive value 68%. Employing a cut point of 250 muMol/L for the second generation assay, the sensitivity to detect diabetes was 81%, specificity 87%, and positive predictive value 43%. However, to detect diabetes and glucose intolerance using the second generation assay, the sensitivity was 39% and specificity was 86%. CONCLUSION: This study demonstrated that a single measurement of either first or second generation fructosamine showed promise as a screening test for diabetes, but not impaired glucose tolerance, in older people. PMID- 8409156 TI - How different are VA nursing home residents? AB - OBJECTIVE: To identify similarities and differences between VA nursing home residents and other nursing home residents. DESIGN: Comparison of cross-sectional data from three sources. PARTICIPANTS: Residents of VA nursing homes nationwide in early October 1986 (n = 10,117); participants in the 1985 National Nursing Home Survey (NNHS) (n = 5,243); residents assessed in New York State nursing homes in 1988 (n = 94,840). MEASURES: Age-stratified comparisons were made between the VA and the NNHS for gender, marital status, race, ethnicity, length of stay, activities of daily living (ADL) status, and selected diagnoses and conditions. Additionally, case-mix data were compared between the VA and the New York State populations. MAIN RESULTS: The population of VA nursing homes is overwhelmingly men (96.1% versus 28.4% in the NNHS), and 31.2% of the VA population is under 65 years of age compared with 11.6% in the NNHS. Young ( < 65) VA residents are considerably more impaired in ADL than young residents in the NNHS; differences are less pronounced in those over 65 years old. VA case mix is slightly higher than the overall New York State population though the distribution of residents into categories in the Resource Utilization Groups, Version II system is somewhat different. CONCLUSIONS: VA nursing homes contain a substantial distinctive population of seriously impaired residents under 65 years of age. Though differences exist, older VA residents have many similarities to residents of non-VA nursing homes and constitute a functionally impaired population that can provide insights into the status of nursing home residents generally. PMID- 8409157 TI - Oral disease and physical disability in community-dwelling older persons. AB - OBJECTIVE: The aim of this epidemiologic investigation was to determine whether poor oral health in older persons was associated with physical disability. DESIGN: The study was a cross-sectional survey involving in-home interviews and dental examinations of older persons. SETTING: A random sample of 68 cities and towns was selected from the six New England states, with stratification according to population size. PARTICIPANTS: The study sample consisted of 1,156 community dwelling (non-institutionalized) individuals aged 70 and over, randomly selected from the Medicare beneficiary lists for each city and town. MEASUREMENTS: Oral health was assessed by three dichotomous indices: edentulism (no teeth); current caries, including either coronal or root decay; and periodontal disease, as measured by gingival pocket depth. Physical disability was indicated by the subject's self-report of difficulty in the areas of personal care (eating, bathing, dressing, and using the toilet) and mobility (walking, bed transfer, getting outside). Additional independent variables included age, sex, number of teeth, education, living alone, oral hygiene practices, and time since last dental visit. RESULTS: We found a direct association between specific areas of physical disability and current caries and edentulism. The risk of poor oral health did not increase with advancing age once the related risk factors were controlled for. Those subjects with mobility disabilities were at increased risk of tooth loss; those with personal care limitations were at increased risk of current caries. CONCLUSIONS: Physical disability should be added to the list of known risk factors for oral disease among the older population. Our findings call attention to the need for health care providers to screen for oral health problems among disabled older persons. Further gerontologic research is needed to identify the mechanisms linking physical disability with oral disease in older persons. PMID- 8409158 TI - Physical Performance Test and Activities of Daily Living scales in the assessment of health status in elderly people. AB - OBJECTIVE: To compare the ability of Basic Activities of Daily Living (BADL), Instrumental Activities of Daily Living (IADL), and the Physical Performance Test (PPT) to detect health status impairments. PATIENTS: Five hundred forty nine community-dwelling elders (89.8% of the eligible elderly population) aged 70 and over; mean age [was] 76.8 +/- 6.1; 179 were males and 370 females. SETTING: City of Ospitaletto, Brescia, Northern Italy. MEASUREMENTS: A multidimensional questionnaire assessing demographic variables, indicators of social activities, psychological function, and somatic health and functional status (BADL and IADL). Also, the PPT was administered. RESULTS: Cognitive and effective status were independently associated with BADL, IADL function, and age; number of drugs were also associated with IADL function. Other health variables (number of diseases, number of symptoms, and global health score) did not independently contribute to explaining the BADL and IADL variance. Cognitive status, number of symptoms, number of diseases, number of drugs, and global health were independently associated with PPT. CONCLUSIONS: Chronic diseases may affect functional status in a manner that is insensitive to traditional self-report ADL and IADL measures. Performance-based measures may capture this impairment before more severe functional loss emerges. PMID- 8409159 TI - Visual field loss in Alzheimer's disease. PMID- 8409160 TI - The biopathology of aging of the endocrine system: the parathyroid glands. PMID- 8409161 TI - Constipation in the elderly. AB - OBJECTIVE: To explore the distinction between true clinical constipation and the subjective complaint of constipation in elderly people and to review the pathophysiology, symptoms, diagnosis, causes, and treatment. DATA SOURCES: A computer-assisted and manual search of the English language literature using MEDLINE 1966-1991, Index Medicus 1988-1992, reference lists of selected articles, and relevant textbooks. STUDY SELECTION: Studies that provide information on lower bowel function and laxative and enema use in the elderly subjects were reviewed. Article selection was not limited by study design. DATA EXTRACTION: Relevant data were abstracted from the results of physiological, cohort and case control studies, and clinical trials. The text discusses the methodological strengths and flaws of these studies and excludes management approaches formulated from uncontrolled clinical observation. RESULTS OF DATA SYNTHESIS: Constipation of the elderly is not well defined in the current literature. Self reported constipation and laxative use increase with age, while a similar escalation in true clinical constipation is not shown. Physiological changes in the lower bowel predisposing toward constipation do not occur with normal aging. Patient selection criteria for studies examining the pathophysiology of constipation differ in their definition of constipation and their inclusion of coexisting chronic illness. Nevertheless, there is consistent evidence for prolonged transit through the sigmoid colon and rectum, especially in frail elderly patients, and reduced rectal tone with impaired sensation, particularly in patients with rectal impaction. Few studies rigorously examine "risk factors" and non-pharmacological interventions in constipation. The results of most laxative trials require cautious interpretation because of inclusion of patients without diagnostically proven constipation, use of combined laxative preparations, and unreliable outcome measures. Certain laxative agents however appear more appropriate for use in elderly people. CONCLUSION: Although the subjective complaint of constipation and habitual laxative use increase with age, the epidemiological data suggest that true clinical constipation does not. Physiological changes predisposing toward constipation are not an inevitable consequence of aging, but appear to be specific to the condition. The available data do not confirm many suspected "risk factors" nor the benefits of commonly used non-pharmacological and pharmacological treatments, but they do provide enough information to formulate a practical approach to constipation in elderly persons. PMID- 8409162 TI - Capacity to complete an advance directive. PMID- 8409163 TI - HMO medical directors' perceptions of geriatric practice in Medicare HMOs. AB - OBJECTIVE: To determine the extent to which health maintenance organizations (HMOs) with Medicare risk contracts utilize geriatricians and selected aspects of "organized" geriatric practice. DESIGN: A telephone interview survey. PARTICIPANTS: Eighty-two percent (64 of 78) of the HMOs with Medicare risk contracts as of June 1991. MEASUREMENTS: Questions to medical directors of the Medicare HMOs on (1) the presence of geriatricians, (2) the roles of geriatricians, and (3) "organized" geriatric practice. MAIN RESULTS: Fifty-three percent of the Medicare HMOs have one or more geriatricians, but only 19% have attempted to recruit geriatricians. Geriatricians provide primary care in 76% of the HMOs with geriatricians and serve as specialist consultants in 61%. Geriatricians are reported to be used actively in 32% of the HMOs that have them. The proportion of HMOs utilizing "organized" geriatric activities ranges from a high of 58% for a general health information questionnaire to a low of 12% for a special form(s) for comprehensive geriatric assessment. While the percentage of HMOs using each of the "organized" strategies is higher for the HMOs with geriatricians than for those without, this is statistically significant for only one strategy--the use of special approaches or formal protocols for problems frequently found in the elderly (P = 0.04). CONCLUSIONS: The perception of Medicare HMO medical directors is that about half of the HMOs utilize geriatricians and that there is evidence of "organized" geriatric practice. However, it appears that geriatricians and many of the elements of organized geriatric practice are used to a much lesser extent than experts recommend. Medicare HMOs must themselves test the various components of organized geriatric practice in order to determine their utility. PMID- 8409164 TI - Clock drawing helps when communication fails. PMID- 8409165 TI - Eva, her roommate, and Hitler. PMID- 8409166 TI - Chronic segmental colitis in association with diverticulosis: a clinical syndrome in the elderly? PMID- 8409167 TI - A case of oculopharyngeal muscular dystrophy in a Bulgarian Jew. PMID- 8409168 TI - Geriatric medicine: a true primary care discipline. PMID- 8409169 TI - Factors affecting physician participation in nursing home care. AB - Physicians generally consider nursing home practice a low priority compared with other aspects of their practices. In order to encourage more enthusiastic physician involvement, negative influences needed to be identified so that corrective ideas can be formulated. Negative factors include low reimbursement, frequent office interruptions, excessive paperwork, and a sense of loss of authority. The problem of quality physician involvement must be viewed from new perspectives. An increase in reimbursement is only part of the solution. Other measures that enhance the professional image and responsibility will increase physician participation more than laws and regulations, which increase physician time commitment without increasing reimbursement. Laws and regulations, through misperception, misinterpretation, and misapplication, can have unintended adverse results. Only creative solutions addressing identified negative factors will cut through the Gordian knot, which prevents enthusiastic and quality medical care for all NH residents. PMID- 8409170 TI - The physics of frailty. AB - The contribution of frailty to human morbidity and mortality is immense. Yet it lacks a conceptual framework. By borrowing from the hypotheses provided by the thermophysicists, frailty can be understood in a way that both interrelates the relationship between form and function and provides medical science with an operational insight that yields clinical benefit. Frailty results where the organism is uncoupled from its environment yielding a break in the forward feedback cycle of stimulus to reaction to growth to increased functional competence to improved response to stimulus. Recognition of the centrality of this interrelatedness to health becomes a key theme for illness prevention and therapy. PMID- 8409171 TI - On frailty: when being no longer implies becoming. PMID- 8409172 TI - A speech from the speechless: deletion of long-term care from the total health care budget. PMID- 8409173 TI - The strange case of an older woman who was cured by being allowed to refuse therapy. PMID- 8409174 TI - Restraints and sudden death. PMID- 8409175 TI - Nighttime sleep and bed mobility among incontinent nursing home residents. AB - OBJECTIVE: To describe sleep and body movement patterns in incontinent nursing home residents for the purpose of determining if the residents require nighttime changing and body repositioning on a 2-hour schedule. DESIGN: Cross-sectional survey. SETTING: Four nursing homes. PARTICIPANTS: 118 nursing home residents. MEASUREMENTS: Over two nights, bedside monitoring equipment recorded wrist activity (as a proxy measure for sleep) and body movements of both the shoulder and hip areas in consecutive 2-minute intervals. Specific outcome measures were: (1) Average duration of a sleep episode, peak duration of a sleep episode, and percent of time in bed asleep. (2) The number of 2-minute intervals in which a large movement (45 degree turn) at the shoulder and hip was noted per hour of recording for each resident. (3) The number of resident-initiated, rather than staff-initiated, large movements at the shoulder and hip that occurred within the same 2-minute intervals. RESULTS: There was large variability in all sleep measures; however, on average, residents slept 66% of the time they were in bed. The distribution of these measures suggests that sleep was punctuated with frequent nighttime awakenings. Thirty-three percent of the incontinent residents demonstrated very low levels of resident-initiated movement at the shoulder and hip. Sixty-six percent demonstrated at least one large movement at the shoulder and hip per hour during periods of sleep as well as during periods of wake. CONCLUSION: The majority of incontinent nursing home residents self-initiate sufficiently frequent movements at both the shoulder and hip so as not to be in need of frequent repositioning by nursing staff. Since the sleep of many of these residents is also characterized by frequent awakenings, incontinent nursing home residents may benefit from a schedule of nursing care at night that considers sleep of equal importance to incontinence care and body repositioning. PMID- 8409176 TI - The nighttime environment, incontinence care, and sleep disruption in nursing homes. AB - OBJECTIVE: To evaluate the association between noise, light, nursing care practices, and nighttime awakenings in incontinent nursing home residents. DESIGN: Cross-sectional survey. SETTING: Four long-term care nursing facilities. PARTICIPANTS: One hundred eighteen incontinent nursing home residents. MEASUREMENTS: Over two nights, bedside monitoring equipment recorded wrist activity, resident bed movements, and environmental noise and light changes in consecutive 2-minute intervals. Changes in sleep and bed movement were compared with changes in noise and light that occurred within the same or proximal 2 minute intervals. Noise and light changes in combination with large resident movement at the hip and shoulder were interpreted as related to incontinence care based on observational measures. Specific outcome measures were: (1) the number of noise and light changes as well as staff care practices that did not wake the resident during periods of consecutive sleep, ie, sleep lasting a minimum of 10 minutes; (2) the number of noise and light changes as well as staff care practices that occurred immediately before or during the 2-minute intervals during which a resident woke from a period of consecutive sleep; and (3) the number of such staff care practices that were related to incontinence care. RESULTS: Noise and light changes associated with both general environmental events and more specific nurse care practices were associated with 50% of all waking episodes of 4 minutes or longer and 35% of all waking episodes of 2 minutes or shorter. The major sources of all noise were traced to nursing staff. Eighty-seven percent of all incontinence care practices were associated with episodes of waking. CONCLUSION: The data reported in this paper document that general environmental noise and incidents of nursing care practices, particularly those related to incontinence care, are responsible for a substantial amount of the sleep fragmentation that is common among nursing home residents. PMID- 8409177 TI - Effects of terodiline on urinary incontinence among older non-institutionalized women. Terodiline in the Elderly American Multicenter Study Group. AB - OBJECTIVE: To determine the effectiveness of terodiline, a drug with calcium antagonist and anticholinergic properties, on the frequency of incontinence in older non-institutionalized women. DESIGN: Randomized, placebo-controlled, parallel-group, double-blind trial. SETTING: Twelve outpatient clinics across the United States affiliated with programs in either geriatrics, gynecology, or urology. PARTICIPANTS: Ninety-eight women, age 60 or older, with symptoms of urge incontinence and self-reported frequency of incontinence four or more times per week and involuntary bladder contractions on dual-channel water cystometry. MAIN OUTCOME MEASURES: Self-reported urinary frequency, urgency, number of incontinence episodes, and number of heavily soaked pads. RESULTS: Eighty-one women, average age 71, completed the trial, 40 in the active drug group, 41 in the placebo group. Incontinence frequency and the number of heavily soaked pads were reduced in the active drug group by 64% and 55%, respectively, and by 21% (P = 0.002) and 9% (P = 0.04) in the placebo group. No patients dropped out due to adverse effects. An intention-to-treat analysis of all 98 patients yielded the same conclusion. CONCLUSION: Terodiline is highly effective in reducing incontinence in older, noninstitutionalized women with urge incontinence. Because of its potential association with polymorphic ventricular tachycardia (torsades de pointes), terodiline must undergo further testing to define its safety before it can be recommended for clinical use in the incontinent geriatric population. PMID- 8409178 TI - Comparison of one-day and three-day calorie counts in hospitalized patients: a pilot study. AB - OBJECTIVE: To determine whether a 1-day calorie count can replace the labor intensive 3-day calorie count commonly, performed in hospitalized patients when estimates of caloric and protein intake are required. DESIGN: Pilot study using prospective, non-concurrent review of medical records. SETTING: Hospital. PARTICIPANTS: Thirty patients (mean age 67 years). RESULTS: Mean 3-day intake (952 +/- 91 calories, 41 +/- 4 g protein) was about half of calculated requirements; first-day intake was similar (918 +/- 116 calories, 40 +/- 5 g protein). The first day had high sensitivity (calories 96%; protein 93%) and positive predictive value (calories 100%; protein 96%). Malnutrition was evident; three-fourths of patients had weights below recommended ranges, and 83% were hypoalbuminemic. CONCLUSIONS: Three-day calorie counts are frequently performed in patients suspected of eating poorly. Results of this pilot study suggest that 1-day calorie counts may be a valid alternative. However, readily available anthropometric and biochemical data may be as good an indicator of inadequate dietary intake. PMID- 8409179 TI - Improving neglected influenza vaccination among healthcare workers in long-term care. AB - OBJECTIVE: To increase utilization of influenza vaccination among long-term-care health workers. DESIGN: Before-after intervention study. SETTING: A 300-bed life care community served by 195 staff employees. INTERVENTION: An educational intervention followed by a Vaccination Fair. MEASUREMENTS: Percent receiving vaccination. A post-intervention survey was used to define staff attitudes towards vaccination. RESULTS: Use of influenza vaccination among staff increased from 8% the previous year to 46% during a Vaccination Fair and to 54% during a similar Fair the next year. CONCLUSIONS: Organized institutional efforts to promote immunization among health-care workers, such as a Vaccination Fair, improves compliance among health-care workers. PMID- 8409180 TI - Cross-cultural cognitive examination: validation of a dementia screening instrument for neuroepidemiological research. AB - OBJECTIVE: Validation of a new instrument for screening dementia, the Cross Cultural Cognitive Examination (CCCE), is described. DESIGN: Criterion and concurrent validation and cross-cultural comparison of a new instrument. PARTICIPANTS: All individuals over the age of 40 in a village in southern Guam participated in a door-to-door survey. Alzheimer's and Parkinson's Disease patients and healthy controls aged 40-90 participated in the US mainland study. MEASUREMENTS: The CCCE was administered to all subjects. Effects of age, language, education, and gender on test performances and social-cultural differences were assessed. Concurrent validation of the test with respect to other well accepted screening instruments was determined. RESULTS: High specificity (> 94%) and sensitivity (> 99%) for detecting dementia were found in Guam and US mainland samples, and these were not biased by differences in gender, linguistic preference, education, or cultural background. Sensitivity and specificity of the CCCE matched or exceeded that of already accepted dementia screening instruments. CONCLUSIONS: These validation studies support the usefulness of the CCCE for identifying patients with generalized dementia, rather than focal types of cognitive impairment, quickly and reliably in cross-cultural neuroepidemiological research. PMID- 8409181 TI - Oral nutritional supplement use in elderly nursing home patients. AB - OBJECTIVES: To determine why elderly nursing home patients receive liquid oral protein supplements, what nutritional assessment is utilized, and whether there is evidence of effectiveness. DESIGN: Retrospective, case control study comparing patients over 65 years of age, at two nursing homes, who were served oral supplements (OS) at least twice daily (n = 56), with a random sample of non supplemented, non-tube fed patients (n = 53). Comparisons included medical diagnoses, medications, morbidity and mortality, weight changes, laboratory test results, and functional and behavioral status. RESULTS: Nursing home patients were begun on OS (median time after admission = 2 months, range = 0-72 months) primarily because of weight loss (71%) and poor appetite (16%). Supplemented patients were below an age-adjusted body weight on admission, unlike controls, and continued to lose weight until OS were started. On OS, weight was slowly regained over 9-10 months in a majority of patients to approximate admission weight. Some patients on OS showed improvement in albumin, total lymphocyte count, cholesterol, or hemoglobin, but too few patients had sufficient lab tests to verify any consistent effect. Mortality was higher in OS patients (8 vs 2, P = 0.057), who were also somewhat older (87.9 vs 84.5 years), but there was no statistical difference in infection or hospitalization rate. MEASUREMENTS: Nutritional assessment in these nursing homes consisted almost exclusively of (1) serial measurement of weight, (2) comparison of weight to (a) "ideal body weight" and (b) previous weight, and (3) a subjective evaluation of food consumption. No other anthropometric evaluations or laboratory tests were conducted for nutritional assessment. CONCLUSIONS: The diagnosis of and intervention in under nutrition in nursing home patients is frequently disorganized. In addition, nutritional assessment, either for screening or for following intervention, is hampered by the lack of convenient and unambiguous assessment tools. OS use is associated with weight gain in many nursing home patients and also improves other nutritional parameters in selected individuals. PMID- 8409182 TI - Predictors of formal home health care use in elderly patients after hospitalization. AB - OBJECTIVE: To prospectively study the incidence of and risk factors for home health care (HHC) use in a cohort of elderly medical and surgical patients discharged from acute care. Although HHC is commonly received by patients in this group, its predictors have not been well studied. DESIGN: Prospective cohort study. SETTING: Medical and surgical wards at a university teaching hospital, followed by 23 Medicare-certified HHC agencies in the study catchment area. PATIENTS: 226 medical and surgical patients aged 70 years and older immediately after discharge from acute care. MEASUREMENTS: HHC initiated within 14 days after hospital discharge, measured by direct review of HHC agency records. RESULTS: The incidence of HHC initiated within 2 weeks post-discharge was 75/226 (34%). The median duration of service was 30 days (range 3-483) with a median of 3 visits per week. Four independent predictors of HHC were identified through multivariate analysis: educational level < or = 12 years (relative risk (RR) 3.3; 95% confidence interval (CI) 1.6 to 6.6); less accessible social support (RR, 1.7; CI 0.9 to 3.1); impairment in at least one instrumental activity of daily living (RR, 1.9; CI, 1.0, 3.4); and prior HHC use (RR, 2.1; CI, 1.2 to 3.6). Risk strata were created by adding one point for each risk factor present: with 0-1 risk factors, 8% used HHC; with two risk factors, 28%; with three risk factors, 45%, with four risk factors, 76%. This trend was statistically significant (P < 0.001). CONCLUSIONS: HHC use is common among elderly patients after discharge from acute care. A simple predictive model based on four risk factors can be used on admission to predict HHC use. This model may be useful for discharge planning and health care utilization planning for the elderly population. PMID- 8409183 TI - Early experience with dobutamine stress testing and cardiac cine-tomographic imaging in the elderly: antianginal effects of nifedipine-GITS. AB - OBJECTIVE: To determine the effects of nifedipine-GITS (GITS = gastrointestinal transport system) on angina and cardiovascular responses to stress-dobutamine infusion, we used ultrafast cine-computed tomography (CT) to assess regional wall motion, myocardial perfusion, and indices of ventricular filling and emptying. DESIGN: Randomized, double-blind placebo-controlled efficacy study after an open label dose titration phase. SETTING: University of California, San Francisco. PATIENTS: Elderly patients (> 60 years; n = 9:8 male, 1 female) with coronary artery disease by history and diagnostic treadmill or coronary angiography. INTERVENTION: After a 3-week open-label dose-titration phase, eight subjects were randomized to receive either placebo or nifedipine-GITS at the highest tolerated dose for 2 weeks, followed by a crossover to the alternate therapy for 2 weeks. One declined because of singulus in the open-label period. MAIN OUTCOME MEASURES: Symptomatic angina relief (frequency and nitroglycerin consumption), dobutamine stress responses (time to ischemia during dobutamine infusions, cardiac output, cardiac ejection fraction, ventricular segmental wall motion, and perfusion as measured by ultrafast cine-CT), and reported adverse effects. RESULTS: When compared with placebo, nifedipine-GITS administration was associated with less frequent angina and nitroglycerin consumption (NS) and significantly decreased systolic blood pressure. Nifedipine-GITS administration also increased resting supine heart rates. Dobutamine infusions increased heart rate, cardiac output, cardiac ejection fraction, and stroke volume and induced angina symptoms. Neither double product at angina nor systolic indices of cardiac function in response to dobutamine differed between nifedipine-GITS and placebo, although heart rate responses were greater during nifedipine. A trend toward increased peak filling rates was seen during dobutamine stress in the nifedipine-administration period. In most subjects (6/8), perfusion and regional wall motion abnormalities were not visualized on regional wall motion abnormalities were not visualized on either rest or stress cine-CT studies. Edema without congestive heart failure occurred frequently during nifedipine-GITS administration. CONCLUSIONS: These data suggest that (1) dobutamine stress can be used to induce cardiac ischemia in elderly patients with coronary artery disease, (2) nifedipine-GITS provides symptomatic angina relief in elderly patients, (3) peripheral edema is frequent in elderly patients on nifedipine-GITS, and (4) ultrafast computed cine-tomography testing can be used to assess ventricular performance, but current methodology may not detect perfusion or wall motion abnormalities during angina. PMID- 8409184 TI - Low circulating levels of insulin-like growth factors and testosterone in chronically institutionalized elderly men. AB - OBJECTIVE: To determine the prevalences of and the associations between hyposomatomedinemia and hypogonadism in healthy young men, healthy old men, and chronically institutionalized old men. DESIGN: Survey with serial blood tests. SETTING: Veterans Affairs nursing home and young and old men living in the community. SUBJECTS: Three groups were studied: healthy young men (20-29 years old, n = 32), healthy old men (59-98 years old, n = 30), and chronically institutionalized old men (59-95 years old, n = 112). MEASUREMENTS: Plasma insulin-like growth factor-I (IGF-I), total testosterone (TT), free testosterone (FT), and plasma insulin-like growth factor-II (IGF-II) were measured. In subjects with low testosterone level, serum luteinizing hormone (LH) was also determined. In a subset of chronically institutionalized old men with low IGF-I, the serum growth hormone (GH) level was analyzed during the first 4 hours of sleep. RESULTS: A low IGF-I level (defined as a value below the lower 2.5 percentile of the comparison group) occurred in 85% of the healthy old men when compared with healthy young men (P < 0.001), in 90% of the chronically institutionalized old men when compared with healthy young men (P < 0.001), and in 26% of the chronically institutionalized old men when compared with healthy old men (P < 0.001). In chronically institutionalized old men with low IGF-I compared with healthy young men, nocturnal peaks of serum GH were < 2 ng/mL in most cases. Low TT (defined as a value below the lower 2.5 percentile of the comparison group) occurred in 86% of the healthy old men when compared with healthy young men (P < 0.001), in 88% of the chronically institutionalized old men when compared with healthy young men (P < 0.001), and in 28% of the chronically institutionalized old men when compared with healthy old men (P < 0.001). The results of FT were similar. In 80% of the institutionalized old men with low TT and FT, the serum LH level was low (< 20 mU/mL). In 53% of the institutionalized old men, the IGF-II level was below the lower 2.5 percentile of the healthy old men (P < 0.001). In both healthy and institutionalized old men, IGF-I and IGF-II levels were significantly correlated to each other (r = 0.6), but neither was significantly correlated to TT or FT. In the institutionalized old men, IGF-I was inversely correlated with age and with a diagnosis of dementia; TT and FT were inversely correlated with age and with the degree of dependency in ADL's. CONCLUSIONS: Compared with healthy young men, most healthy old men have low serum IGF-I, TT, and FT levels. The geriatric hyposomatomedinemia and hypogonadism are more severe in institutionalized old men. In the latter group, both endocrine deficiencies are usually of central origin, but their occurrences are not significantly associated. Healthy old men usually have a low level of IGF-I compared with healthy young men, but a similar level of IGF-II; institutionalized old men are usually low in both values. PMID- 8409185 TI - Oncogenic osteomalacia associated with metastatic prostate carcinoma: case report and review of the literature. PMID- 8409186 TI - Diverticular disease in the elderly. PMID- 8409187 TI - Dietary restriction and aging. AB - The diet-restricted rodent model has been and is a major tool in experimental biogerontology. A spectrum of findings indicates that dietary restriction retards the aging processes of mice and rats, the most salient of which is the increase in mortality rate doubling time. It also maintains many physiological processes in a youthful state and, most strikingly, retards or prevents almost all age associated disease processes. Current emphasis is on the mechanisms underlying the anti-aging actions of dietary restriction. The major effort for determining mechanism has focused on putative primary aging processes. A clue has emerged from the findings that it is the restriction of energy intake that is the dietary factor responsible for the anti-aging actions. However, reducing the metabolic rate is not involved. The challenge is to learn how the reduction of energy intake per animal (not per unit of body mass) is coupled to the retardation of aging processes. One of our working hypotheses is that dietary restriction alters nervous and/or endocrine functions that influence the characteristics (not the rate) of fuel use; this modulation in fuel-use characteristics is proposed to retard the aging processes. Our findings on carbohydrate metabolism are in accord with this view. Diet-restricted rats can use carbohydrate fuel as effectively as ad libitum fed rats while maintaining lower plasma glucose and insulin level. Maintenance of these low levels may protect against long-term damaging actions of these substances. Dietary restriction also protects against oxidative damage and, of course, oxidative damage is probably an inevitable component of fuel use.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409188 TI - A short story. PMID- 8409189 TI - Vision of opportunity: clinical optometric research in the VA. PMID- 8409190 TI - Total quality improvement (TQI) for optometric practice in the Department of Veterans Affairs (VA) PMID- 8409191 TI - VA residency programs in optometry: characteristics of the applicants. AB - BACKGROUND: Residency programs in optometry have become an important part of the training of doctors of optometry. Although there have been many studies on the subject, there are no known reports which concentrate solely on the characteristics of residency programs' applicants. METHODS: Applicant data obtained from the application and matching services utilized by VA programs for the 1991-92 and 1992-93 residency years was analyzed and, in certain circumstances, compared against appropriate national data. RESULTS: The ratio of applicants to available residency positions exceeded 2:1. The majority of applicants were women. During each residency application year studied, at least one applicant came from each of the accredited schools and colleges of optometry. Stronger scores from the National Board of Examiners in Optometry and higher grade point averages from the optometry curriculum correlated with the matching of an applicant to a residency program. CONCLUSIONS: As the practice of optometry continues to evolve, the profile of many aspects of optometric education, including residency training, will also change over the years. This paper, as baseline information, may serve in future analyses to demonstrate these changes with respect to applicant characteristics. PMID- 8409192 TI - Superior quadrant visual field loss secondary to temporoparietal craniectomy for brain abscess. AB - BACKGROUND: Visual field defects occur from a variety of causes. As a result, the differential diagnosis of visual field defects and their etiology is important. METHODS: A 64-year old white male was examined in follow up related to the surgical removal of a right temporal lobe abscess secondary to a childhood right side mastoidectomy. The patient underwent ophthalmic examination, which suggested possible changes in his field of vision, and neurobehavioral evaluation related to his visual field defect, a left homonymous superior quadrant defect. RESULTS: Computed tomography (CT) scan of the head without contrast enhancement confirmed the ophthalmic diagnosis and correlated with the patient's medical history. However, neurobehavioral evaluation revealed a number of deficits that were believed to be associated with alcohol abuse. CONCLUSIONS: Visual field loss with changing vision symptoms and a complex history can confound the differential diagnosis of visual field defects. Frequently careful and complete evaluation can confirm and reveal subsequent diagnoses. PMID- 8409193 TI - Central serous chorioretinopathy complicated by subsequent occipital lobe infarction and sinusitis in a drug abuser. AB - BACKGROUND: The presence of multiple occurring, simultaneously complicated clinical findings can be challenging. METHODS: A 39-year-old white male presented initially with central serous chorioretinopathy. Later, the patient reported a great deal of pain over his left eye with nasal vision loss. Comprehensive ophthalmic evaluation and neurologic consultation were invaluable in the diagnosis and management of the patient. RESULTS: Neurologic consultation along with computed tomography (CT) of the head with and without contrast enhancement along with magnetic resonance imaging (MRI) with and without gadolinium revealed lesions in the left occipital lobe, right cerebellum, and left thalamus area. In addition, bilateral maxillary and ethmoidal sinusitis were observed. CONCLUSIONS: Multiple events affecting vision are often related, sometimes not. However, co existing factors can confound and obscure accurate diagnoses. Careful patient clinical and laboratory investigations are frequently necessary to elucidate correct diagnosis. PMID- 8409194 TI - Nonrhegmatogenous retinal detachment in Goodpasture's syndrome: a case report and discussion of the clinicopathologic entity. AB - BACKGROUND: Goodpasture's syndrome is a rare, rapidly fatal disease that consists of glomerulonephritis and pulmonary hemorrhage. Although not common, the ophthalmic manifestations of this disease include hemorrhagic and/or exudative retinopathy and infrequently, nonrhegmatogenous retinal detachment. METHODS: A 25 year-old white male was examined with a complaint of bilateral intermittent blurring of vision. His medical history revealed a diagnosis of Goodpasture's syndrome with subsequent renal failure and associated complications. Ophthalmoscopy and visual field testing demonstrated bilateral, inferior nonrhegmatogenous retinal detachments. RESULTS: Optimum control of both blood pressure and metabolic processes were recommended to increase the chance of resolution of the nonrhegmatogenous retinal detachments. CONCLUSIONS: Although infrequently seen in Goodpasture's syndrome, nonrhegmatogenous retinal detachment may have a good prognosis for recovery when diagnosed early and with treatment of the underlying disease and its metabolic derangements. PMID- 8409195 TI - Support for the vasogenic theory of glaucoma: case reports and literature review. AB - BACKGROUND: The influence of systemic disease, vascular abnormalities, hematologic and rheologic factors, pulsatile ocular blood flow, perfusion pressure, autoregulation, optic disc hemorrhage and the results of optic nervehead fluorescein angiographic studies are reviewed in the context of supporting evidence for the vasogenic theory of glaucoma. METHODS: Two case reports of patients with confirmed carotid artery occlusive disease and one case report of a patient with a history of hypovolemic blood loss are presented. RESULTS: These cases illustrate the importance of hypotensive crisis, perfusion pressure, collateral blood flow and the optic nerve head vascular autoregulatory mechanism in the pathogenesis of glaucoma and the differential diagnosis of pseudoglaucoma. CONCLUSIONS: Evaluation and consideration of both mechanical and vasogenic causes of the glaucomas is essential in the management of the glaucomas. It is also necessary to differential diagnose the "pseudo-glaucomas," and to manage them correctly. PMID- 8409196 TI - Sphenoidal mucocele with cavernous sinus invasion. AB - BACKGROUND: Intracranial extension of a sphenoidal mucocele may be marked by persistent headache, nerve palsies, and total or partial vision loss. METHODS: A 74-year-old white male was examined with right frontotemporal headache and constant diplopia. Previously, he had noticed a droop in his right eyelid. RESULTS: Careful ophthalmic examination and laboratory studies, including computed tomography revealed a sphenoid mucocele with complications. As a result, the patient underwent a sphenoidectomy. CONCLUSIONS: The evaluation of patients with acute onset of multiple eye and vision problems can often lead to a significant neurologic diagnosis. PMID- 8409197 TI - Estimation of total DNA in crude extracts of plant leaf tissue using 4',6 diamidino-2-phenylindole (DAPI) fluorometry. AB - A method of estimating total double-stranded DNA in crude extracts of citrus leaf tissue by evaluating the enhancement of fluorescence intensity of 4',6-diamidino 2-phenylindole (DAPI) was assessed. For pure citrus DNA and citrus leaf tissue crude extract each in the presence of 100 ng/ml DAPI, excitation spectral response curves converged at excitation wavelength of 360 nm. At this excitation wavelength, maximum fluorescence intensity occurred across a range of emission wavelengths from 445 nm to 460 nm. The appropriate excitation and emission wavelengths were shown to be 360 nm and 450 nm, respectively. Fluorescence intensity increased linearly with DNA concentration and non-DNA components of the tissue homogenates had negligible effect on fluorescence at these wavelengths. Sodium dodecyl sulfate (SDS) in the incubation solution resulted in some suppression of DAPI-DNA fluorescence and produced a non-linear response to changing DNA concentration. The method should be applicable to DNA quantitation from crude tissue extracts of any plant species. PMID- 8409198 TI - Bead-model calculation of scattering diagrams: Brownian dynamics study of flexibility in immunoglobulin IgG1. AB - We consider the calculation of the angular dependence of the scattering intensities for models composed of spherical elements of arbitrary size, in which some of the spheres may have a size close to that of the whole particle. This minimizes the number of spheres and makes it possible to use the same bead models for the prediction of scattering diagrams and hydrodynamics properties. A simple expression is employed for the scattering intensity. Particularly, we discuss the validity of some versions of the Debye scattering equation for bead models. The method could be used for any macromolecule either rigid or with conformational variability. The application of the method to a simple model for IgG1 shows that influence of flexibility in the scattering curve is stronger by the end of the first decade of decay and also shows that the range of linearity in the Guinier region is not a good method to characterize flexibility between arms, because it is very likely that experimental errors will hide the small differences between the extreme cases appreciated in our calculations. PMID- 8409199 TI - The interference of cysteine thiols in the detection of glycated and non-glycated proteins by modified silver staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - In the previous study a staining intensification of in vitro glycated collagen type I versus a non-glycated one after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under a modified silver staining procedure was observed (Hodny, Z., Struzinsky, R. and Deyl, Z. (1992) J. Chromatogr. 578, 53 62). While investigating the specificity of this stain to glycation product(s) on protein we have observed that a great number of proteins (e.g. bovine serum albumin) was sensitive to this stain even in a non-glycated state. It is proposed from results of the analysis of amino acid composition of these proteins that their better stainability correlates with the amount of cysteine present in the protein. Modification of SH groups by iodoacetamide (or N-ethylmaleimide) had an inhibitory effect on the staining of bovine serum albumin (and some other proteins) in its 'native' state but had no visible inhibitory effect on their staining in the glycated state. However, the positive staining response of a great number of components from cellular lysates even after iodoacetamide treatment indicates the existence of further chemical groups (either of protein or nucleic acid origin) participating in this silver staining method. PMID- 8409200 TI - Di- and tetracarboxylate derivatives of pyridines, bipyridines and terpyridines as luminogenic reagents for time-resolved fluorometric determination of terbium and dysprosium. AB - A group of compounds composed of pyridine, 2,2'-bipyridine and 2,2':6',2'' terpyridine as the energy absorbing, triplet sensitizing moieties and two or four carboxylic acids as the chelating groups were synthesized and their ability to enhance terbium ion luminescence was elucidated. The dicarboxylic acid derivatives form highly luminescent Tb3+ chelates with luminescence quantum yields approaching 50%. Additional groups, such as methoxy groups or additional pyridine rings in the aromatic structure can be utilized to shift the excitation maxima towards longer wavelengths in order to produce more practicable excitation for practical use. The ligands giving the highest Tb3+ emissions were further evaluated in a two-step fluorescence enhancement system constructed for specific binding assays based on the simultaneous use of three or four ions as the labels (Eu3+, Tb3+, Sm3+ and Dy3+). In the two-step procedure, the first step comprises dissociative fluorescence enhancement based on an acidic solution of an aromatic beta-diketone used as the luminogenic ligand, whereafter Eu3+ and Sm3+ are quantitated. In the second step, the lanthanide ions form new chelates with the added ligands. These ligands are able to transfer the excitation energy they have absorbed with high efficiency to the chelated ions and produce high ion luminescence for Tb3+ and Dy3+. PMID- 8409201 TI - A radio-enzymatic method for the estimation of L-leucine-specific radioactivity. AB - A method is presented for the estimation of L-leucine concentration and radioactivity in biological samples. The sample L-leucine is specifically bound to tRNA and its radioactivity estimated in the presence of either added labelled L-leucine or cold L-leucine (in the same proportion), as well as in the presence of a large excess of cold L-leucine. The latter gave the measurement of non leucine radioactivity present in the sample. The measurements in the presence and absence of labelled/cold L-leucine allowed the estimation of L-leucine levels and radioactivity by using a simple set of calculations and a standard curve built with known cold L-leucine concentrations in the presence of a fixed known amount of [14C]L-leucine. PMID- 8409202 TI - Method to improve the accuracy of membrane osmometry measures of protein molecular weight. AB - Membrane osmometry provides a simple method to determine protein molecular weight but accuracy is limited by nonideal behavior. Recent studies (Fullerton et al., Biochem. Cell Biol., in press) show that non-ideal osmotic response of protein solutions is described by the empirical equation, Msv/M(s) = RT rho/A(s) x 1/II+I, where M(s) = mass of solute, Msv = mass of solvent, R = the Universal gas constant, T = absolute temperature, rho = solvent density, A(s) = solute molecular mass, II = osmotic pressure, and I = the nonideality parameter. This linear relation is used in this paper to demonstrate that measurement of molecular weight from the slope simplifies such measures and improves the accuracy relative to classical methods. The molecular weight of bovine serum albumin is measured with error less than 0.9%. The single dimensionless non ideality parameter, I = 4.05 + 0.07, describes non-ideal curvature in the typical IIV = nRT diagram better than the customary second power viral expansion requiring 3 fitting constants. Analysis of eight data sets on four proteins from the literature shows that molecular weight calculated from the slope of the new equation agrees with chemical molecular weight within an RMS error of only 1.9%. PMID- 8409203 TI - Simple measurement of glycosaminoglycan produced by cultured fibroblasts using 4 methylumbelliferyl beta-D-xyloside. AB - A simple and rapid method was devised for measurement of glycosaminoglycan produced by cultured cells. 4-Methylumbelliferyl-beta-D-xyloside was added to the medium of the cultured cells. After incubation, glycosaminoglycan, which was produced from 4-methylumbelliferyl-beta-D-xyloside as a primer and secreted into the medium, was separated by proteinase digestion, trichloroacetic acid treatment and ethanol precipitation. The glycosaminoglycan, bearing a fluorescent moiety at the reducing terminal, was electrophoresed on cellulose acetate membrane, and then the fluorescent band visible on the membrane was extracted. The fluorescence of the band was measured, and from this the amount of glycosaminoglycan was estimated. Using this method, it was possible to quantify a very small amount of glycosaminoglycan with relatively high sensitivity without employing a radioisotope. This method was applied for determination of glycosaminoglycan produced by cultured fibroblasts from human uterine cervix, and also the effect of a hormone on glycosaminoglycan production. It was found that uterine cervical fibroblasts produced twice as much glycosaminoglycan as skin fibroblasts. PMID- 8409204 TI - Conformational studies on some Cl'-branched beta-D-nucleosides by 1H-NMR spectroscopy and molecular mechanics calculations. AB - Solution structures of 1-(beta-D-psicofuranosyl)thymine (HMT) (5) and 1-(1'-cyano beta-D-ribofuranosyl)-thymine (CNT) (6) based on 3JH,H coupling constants (1H-NMR at 500 MHz) and nOe enhancement studies were further refined by molecular mechanics calculations using the AMBER force field. These complementary NMR molecular mechanics studies helped us to define the torsion angles inside the NMR defined conformational hyperspace. Atom-centered monopole charges were derived for molecular mechanics calculations by fitting the molecular electrostatic potential on freely optimized geometries of 5 and 6 using HF/3-21G level of theory by GAUSSIAN 92 program. The conformation of 5 and 6 can be summarized as follows: (i) the pseudorotational analyses showed that the North conformer is predominant in both 5 (> 70%) and 6 (97%). While the molecular mechanics could correctly predict the energetic preference of North over South type puckered sugar moiety for 5 and 6 it could not provide any clue to the fact that the 1'-CN group in CNT (6) drives the pseudorotational equilibrium more effectively towards North than 1'-CH2OH in HMT (5). (ii). The NMR-observed preference of anti over syn conformation across the glycosyl bond in 5 and 6 was correctly shown by the energetic preference of anti conformers by approx. 10 kJ/mol. (iii) The populations of the staggered rotamers across C4'-C5' (gamma +, gamma t and gamma ) calculated from 3J4'5' and 3J4'5" coupling constants from NMR spectroscopy show that gamma + and gamma t rotamers are preferred. Molecular mechanics calculations are also in an excellent agreement here with the results of solution studies: in 5 gamma + and gamma t rotamers are equally populated and a small energy difference in potential energy is established, while in 6 the larger energy difference in potential energy is found which reflects a higher preference for gamma + rotamer in solution. The energetic preferences found among the lowest energy conformers of 5 (North-gamma +/gamma t-anti-epsilon t1) and 6 (North-gamma +/gamma t-anti) in molecular mechanics calculations are in an excellent agreement with the preferences of the major conformers found by NMR spectroscopy in solution. PMID- 8409205 TI - Thiol and disulfide determination by free zone capillary electrophoresis. AB - A rapid, sensitive and simple method for the determination of reduced and oxidized glutathione, cysteine, cystine, cysteamine, cystamine and their respective mixed disulfides is described. The compounds were separated and identified in a single step by capillary zone electrophoresis. The method was used to follow the thiol-disulfide interconversion and to measure glutathione levels in lens extracts. PMID- 8409206 TI - Radioiodination of transforming growth factor-beta (TGF-beta) in a modified Bolton-Hunter reaction system. AB - We have optimized the use of the Bolton-Hunter reagent to prepare 125I-labeled transforming growth factor-beta (TGF-beta). Conditions were developed to obtain monovalent modification of human-recombinant TGF-beta 2 (hrTGF-beta 2) at a basic pH necessary for efficient protein acylation (> or = 26% of theoretical) while obviating the problems of TGF-beta aggregation/precipitation. The purified Bolton Hunter labeled hrTGF-beta 2 had a specific activity of 1.8-2.1 microCi/pmol, and the 125I label was fully acid-precipitable. [125I]hrTGF-beta 2 was electrophoretically indistinguishable from unlabeled starting material and displayed full immunoreactivity with polyclonal anti-TGF-beta 2 antibody. Both hrTGF-beta 2 and Bolton-Hunter-labeled [125I]hrTGF-beta 2 inhibited the growth of mink lung epithelial cells with equal efficacy. These data validate a modified conjugation-iodination method for TGF-beta and invite general use of the Bolton Hunter reagent for iodination of other TGF-beta isoforms and peptides similarly susceptible to precipitation/aggregation under standard Bolton-Hunter incubation conditions. PMID- 8409207 TI - Activity assays for characterizing the thrombolytic protein fibrolase. AB - Characterization of the thrombolytic agent fibrolase was accomplished employing specific proteolytic and thrombolytic assays. This paper describes a method to measure enzyme proteolytic activity using the oxidized beta-chain of insulin as a substrate. Advantages of this method include a short incubation time for substrate cleavage followed by an isocratic HPLC method with a retention time of approx. 5 min. Proteolytic activity can be rapidly and easily quantitated with this procedure. An azocasein assay was also used to quantitate proteolytic activity. This method was optimized with respect to substrate concentration and incubation time allowing for the rapid quantitation of fibrolase activity. A thrombolytic assay is described which employs fibrin plate clearance and has the advantage of rapid and accurate quantitation compared with previously described methods. It also allows for the standardization of fibrolase in plasmin equivalent units. PMID- 8409208 TI - Measurement of free Zn2+ ion concentration with the fluorescent probe mag-fura-2 (furaptra). AB - The fluorescent probe mag-fura-2, previously used to measure [Mg2+], can also be used to measure [Zn2+]. The peak in the excitation spectrum occurs at 323 nm for Zn2+, compared with 335 nm for Ca2+ and Mg2+. This allows simultaneous measurements of [Zn2+] and either [Ca2+] or [Mg2+], by using 3 excitation wavelengths. The dissociation constant for Zn2+ is 20 nM at pH 7.0-7.8, ionic strength 0.15 and 37 degrees C. This allows [Zn2+] to be measured in the range from 0.5 nM to 1 microM. Mag-fura-2 was used to measure [Zn2+] in Zn2+/albumin and Zn2+/histidine mixtures in a physiological buffer at 37 degrees C and pH 7.4. The data obtained enable one to formulate Zn(2+)-buffers for the 1 to 100 nM Zn2+ range. PMID- 8409209 TI - The tetramethylammonium chloride method for screening of cDNA libraries using highly degenerate oligonucleotides obtained by backtranslation of amino-acid sequences. AB - We describe a method for screening of cDNA libraries with highly degenerate oligonucleotides using tetramethylammonium chloride (TMAC). This method is a convenient alternative to using probes generated by the polymerase chain reaction (PCR), especially when these cannot easily be made. Nylon filters were prehybridized in buffered sodium chloride and hybridized with labelled oligonucleotide in buffer containing 3 M TMAC. In TMAC the melting temperature of the oligonucleotide is independent of the G + C content, thus only depending on the length. This was confirmed by the cloning of 13 specific cDNAs with G + C contents between 27% and 61% using 15- to 20-mer oligonucleotides with a degeneracy up to 512. The method was further improved for highly degenerate oligonucleotides by testing hybridization of four 18-mer oligonucleotides, each containing one deoxyinosine (I) instead of A, G, C or T, respectively. Oligonucleotides containing I pairing with A, G or T may have slightly lower melting temperatures than those pairing with C. At practical circumstances all oligonucleotides hybridize about equally well at hybridization temperatures 10 degrees C below the irreversible melting temperature. This was further confirmed by the cloning of four cDNAs with oligonucleotides containing deoxyinosines at 3 or 4 ambiguous positions. PMID- 8409210 TI - PCR to identify specific clones of interest for DNA sequencing. AB - We describe a rapid method for identifying specific clones of interest for the purpose of sequencing. The method essentially is polymerase chain reaction using one internal primer and one vector specific primer. The procedure is particularly useful when relatively large numbers of clones are to be examined either to establish the nucleotide sequence of a full-length cDNA or to find a specific section of a large DNA. The relative orientations of inserts in different clones can also be determined using the same procedure. PMID- 8409211 TI - Ketoisocaproate contamination errors in protein synthesis determinations using L[1-14C]leucine. AB - Protein synthesis rate determinations in vivo using L-[1-14C]-leucine may be underestimated because of contamination by radioactive ketoisocaproate (KIC) resulting from leucine metabolism. The aim of this work was to set up a reliable method to determine the KIC/leucine radioactivity ratio in protein-free homogenates, and to apply it to study the extent of the protein synthesis ratio error due to KIC contamination. Cation-exchange chromatography using Dowex AG 50W X8 resin was used to separate KIC from leucine, eluting KIC with water and leucine with 4 M ammonia. The errors found in the protein synthesis ratio were 6.20% in liver and 2.34% in jejunum. PMID- 8409212 TI - Quantitative and qualitative characterization of virus envelope proteins and specific polyclonal antibodies. AB - A method for simultaneous qualitative and quantitative characterization of virus envelope proteins and virus-neutralizing antibodies is described. The quantitative estimate is based solely on the precipitation of the antibody/antigen (Ab/Ag) complexes at equivalence under equilibrium conditions. The qualitative analysis is performed by determining the diffusion coefficients, i.e., molecular masses of antigen and antibodies. Isolation, purification, labelling of antigen and antibodies and use of any standards are not required. The analyses are carried out directly in crude biological fluids in which antigen and antibodies naturally occur. The results obtained are independent on the avidity of the Ab/Ag system to be analysed. The method was tested by using antigenic subunits of Newcastle disease virus (NDV) and four chicken anti-NDV immune sera of different avidities. PMID- 8409213 TI - Cooling rates of tissue samples during freezing with liquid nitrogen. AB - The time-course of temperature changes undergone by tissue samples during freezing in liquid nitrogen has been studied with small thermocouple probes. The specific heat of liver, muscle, adipose tissue and blood has been measured with a calorimeter. Most tissues have lower specific heat values than water. The lower thermal conductance of adipose tissue and fat muscle result in much longer freezing times. It may take an inordinate amount of time for tissue samples to freeze when immersed in liquid nitrogen; in addition, not all tissues behave in the same way, so it takes longer for a muscle sample to freeze than for a piece of liver. These differences between tissues are considerable and may affect the outcome of experiments if not taken into account. PMID- 8409214 TI - Sympathetic and parasympathetic neuromuscular junctions in the guinea-pig sino atrial node. AB - The structure and organization of cholinergic and adrenergic varicosities in the sino-atrial node of the guinea-pig heart was determined by electron microscopy. When random sections of tissue were examined, some varicosities were found in close proximity (less than 90 nm) to a muscle cell, while others appeared to be some distance (greater than 90 nm) from the nearest muscle cell. When the organization of individual varicosities and their relationships with nearby cardiac muscle cells were determined by examining serial sections of tissue, it was found that most varicosities which lost all or part of their Schwann cell wrap formed close appositions with one or more cardiac muscle cells. At the regions of close apposition, the neuromuscular clefts were filled with a single layer of basal lamina, giving neuromuscular separations of about 80 nm. Although evidence of pre-synaptic or post-synaptic thickenings was not found, there was an accumulation of synaptic vesicles towards the regions of close apposition. These observations are discussed in relation to the idea that in a number of different tissues, most autonomic varicosities which lose part of their Schwann cell wrap, form organized neuromuscular junctions and that these junctions may be the sites of neuromuscular transmission. PMID- 8409215 TI - Influence of peri-arterial hepatic denervation on the glycemic response to exercise in rats. AB - Exercise is known to increase hepatic glucose production. Previous studies have suggested that the sympathetic nerves only marginally contribute to this process. This study examined whether increased catecholamine response or increased adrenoceptor sensitivity might have affected previous results showing no effect of hepatic denervation on the increased hepatic glucose production during exercise. Hepatic sympathetic denervated rats, sham-operated rats and control rats were forced to swim against a counter current for 15 minutes. Denervations and sham operations were performed 9 days prior to swimming. The results show that denervation did not affect the changes in levels of blood glucose, plasma FFA, and catecholamines before, during and after swimming. Furthermore, hepatic adrenoceptor sensitivity was not altered in denervated rats, since intravenous infusions of epinephrine (20 ng/min) and norepinephrine (50 ng/min) similarly changed blood glucose and plasma FFA levels in liver-denervated, sham-operated and control rats. Thus, the increase in blood glucose levels during intravenous infusion of epinephrine and norepinephrine in the respective groups was 1.2 +/- 0.3 and 1.0 +/- 0.3 mmol/l (liver-denervated rats), 1.6 +/- 0.4 and 0.7 +/- 0.3 mmol/l (sham-operated rats) and 1.3 +/- 0.3 and 0.8 +/- 0.3 mmol/l (control rats), respectively. After adrenodemedullation, however, the rise of glucose levels during swimming in liver-denervated and control rats was completely abolished. Thus, the glucose response to swimming with and without adrenodemullation was 0.1 +/- 0.4 and 1.7 +/- 0.4 mmol/l in liver-denervated rats (P < 0.01) and -0.2 +/- 0.4 and 2.2 +/- 0.2 mmol/l in control rats (P < 0.001), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409216 TI - Effects of atropine on the responses of rapidly adapting pulmonary stretch receptors and dynamic lung compliance to sodium cyanide-induced hyperpnea. AB - The responses of tracheal pressure (PT) and heart rate (HR) to electrical stimulation of the peripheral cut-ends of the vagus nerves (10-15 V, 10 Hz, 1 ms) to activate both myelinated and non-myelinated fibers were examined before and after administration of atropine (1 mg/kg) in artificially ventilated, bilaterally vagotomized rabbits. Vagal stimulation caused an increase in PT and a decrease in HR. The PT response to vagal stimulation was completely blocked by atropine which significantly reduced the bradycardia evoked by the stimulation. In other series of experiments, we also examined the responses of rapidly adapting pulmonary stretch receptors (RARs) and dynamic lung compliance (Cdyn) to intravenous injections of sodium cyanide (NaCN, 20 and 30 micrograms/kg) before and after administration of atropine (1 mg/kg) in spontaneously breathing rabbits with vagal efferent activities. Administration of NaCN led to an increase in RAR activity associated with a decrease in Cdyn, and these two effects became more prominent by increasing the dose of NaCN. Atropine treatment did not significantly alter the responses of RARs and Cdyn to the injections of NaCN with different doses. These results suggest that the changes of RAR activity and Cdyn produced by hyperpnea due to NaCN administration are not influenced by the vagally mediated bronchoconstriction. PMID- 8409217 TI - A low frequency, high amplitude rhythmic fluctuation of laser-Doppler skin blood flow after subarachnoid phenol block. AB - A 51-year-old male with a huge chondrosarcoma received subarachnoid dorsal root blocks with 10% phenol in glycerine to treat severe pain along the left leg. The dermatomes below the Th9 lost all somatic sensation on the left side after the nerve blocks, but the patient was not completely relieved from the pain. Laser Doppler flowmetry on the toe of the left foot disclosed an increased blood flow and an abnormal fluctuation of the cutaneous capillary blood flow, i.e. a high amplitude rhythmic (HAR) wave with 2.5 to 3 cycles.min-1. The low frequency HAR wave persisted for the subsequent 3 months until a tingling sensation returned to the left leg. It would seem that some travelling roots of the sympathetic nerve were preserved from the chemical neurolysis and the remaining efferent and afferent nerve fibers were responsible for the persisting low frequency HAR wave and pain in the left leg. PMID- 8409218 TI - Muscle sympathetic outflow in Buerger's disease. AB - One of the pathophysiological features in Buerger's disease, i.e. thromboangiitis obliterans (TAO), contains a sympathetic contribution which relates to vasospastic phenomenon. The purpose of this study is to clarify sympathetic mechanisms in TAO. Muscle sympathetic nerve activity (MSA) was recorded using a microneurographic technique in patients suffering from TAO. The cold pressor test (by immersing the hand of the subject into ice cold water) was used as a test to examine the sympathetic responsiveness to local cold stimulus. The basal level of the MSA in the TAO group was lower than that in the control group (control group vs. TAO group: 39.8 +/- 12.8 bursts/min vs. 26.1 +/- 12.1 bursts/min in burst rate, P < 0.05, 61.8 +/- 8.3 bursts/100 heart beats vs. 39.5 +/- 22.3 bursts/100 heartbeats in burst incidence, P < 0.05). The responsiveness of MSA to the cold pressor test in the TAO group was higher than that in the control group (TAO group vs. control group: 385 +/- 191% total MSA vs. 198 +/- 156% total MSA, P < 0.05). We directly demonstrated higher responsiveness of MSA to the local cold stimulus in patients with TAO. This hyperresponsive MSA might contribute to the pathophysiologic feature in TAO. PMID- 8409219 TI - Respiration-related features of sympathetic discharges in the developing kitten. AB - The objectives of the present study were to identify sympathetic discharge features, either spontaneous or evoked, which exhibited maturational changes. The spontaneous sympathetic nerve discharge of all kittens was comprised of tonic activity as well as phasic inspiratory activity; the latter occurred synchronously with the C5 phrenic discharge. Sympathetic activity during inspiration was depressed (inhibited or disfacilitated) by vagal afferent inputs as was indicated by the marked increases of amplitude following vagotomy. Hypoxia (10% O2) elicited changes in the amplitude of the sympathetic inspiration-related discharge which were identical to those of the phrenic; discharge amplitudes were either increased, or decreased, or increased and then decreased (i.e. biphasic response). Regardless of the change in sympathetic inspiration-related activity during hypoxia, the amplitude of tonic activity during expiration was often increased. Such changes in the discharge characteristics of the cervical sympathetic nerve in response to vagotomy or hypoxia were not dependent upon maturational processes as they were similar to those of adult cats. However, an age-dependent phenomenon was revealed by autopower spectral analyses which showed that the phrenic and sympathetic nerve activities in animals > or = 45 day old were modulated by inputs from a common source because their coherence spectra exhibited correlated periodicities in the 7-10 Hz range. PMID- 8409220 TI - The application of high-definition video systems in medicine. AB - High-definition television is now available in two quite separate systems, with advantages over existing broadcast television formats which would benefit the medical user. The 1125 line, 60 Hz system is described, and the technical jargon explained. Production and broadcast techniques are discussed. Some recent applications of high-definition television for the demonstration of medical procedures are described, and possible future developments for medical users are outlined. A glossary of technical terms is included for those not familiar with video technology. PMID- 8409221 TI - Stereoscopic imaging and its role in medical illustration. AB - Stereoscopic imaging is becoming more widespread as developments in computer software and hardware allow for rapid manipulation of electronically gathered medical data, and of computer-generated models. This paper examines the current three-dimensional multimedia scene, evaluates its place in training, and examines stereoscopic imaging and viewing techniques as well as the availability of stereoscopic equipment. PMID- 8409222 TI - The Macalister archive: records from the Queen's Hospital, Sidcup, 1917-1921. AB - The Queen's Hospital opened in 1917 to care for soldiers receiving facial injuries in Western Front trenches, usually as a result of a gunshot wound. Some 8000 patients were treated by the medical teams of the UK, the Dominions and the USA. The wartime records were removed by their respective sections in 1921, but Queen Mary's Hospital has recently reacquired those of the New Zealand section, rescued from imminent destruction by Professor A.D. Macalister, late Dean of the Dental School at Dunedin, and kindly donated by him. There are 282 sets of case notes containing typescript summaries, clinical photographs and radiographs, drawings, 77 watercolor paintings and a life-size wax model of head and upper torso illustrating some of the surgical techniques. The archive is a fine example of medical illustration 75 years ago, and provides invaluable detail on the plastic surgery and dental reconstructive methods that were developed at Sidcup. PMID- 8409223 TI - Videodisc and compact disc technologies. PMID- 8409224 TI - Illustrations from prisoner-of-war camps on the Japanese Thai-Burma Railway Project. PMID- 8409225 TI - The management of serial photographs in a small research project. AB - A research project in plastic surgery which removed tattoos by deep shaving required serial photographs to illustrate the comparative efficacy of different antifibroblast creams. To obtain reasonably consistent results it was necessary to establish a set of guidelines which would fulfil the aim. This paper describes the photographic technique, the equipment used, and management of the project using a database. PMID- 8409226 TI - Early days in the St. Mary's Medical School Photographic Department: Part 2. AB - Following Part 1 (1992; 15: 133-137), this second article covers the 1950s and 1960s. The enormous amount of work done for the Medical School's Centenary Exhibition to which no visitors came is outlined: Sir Alexander Fleming's picture of the 1928 culture plate of Penicillium; development of the cinelaryngeal camera; the introduction of modern techniques of duplication. Some comments on the quality that is lost with the use of modern automated 'foolproof' equipment. PMID- 8409227 TI - The Trust experience. AB - Since the introduction of 'The NHS and Community Care Act' in 1990, the National Health Service and the role of medical illustrators within it have undergone many changes. Following the new Act, a number of hospitals became National Health Service (NHS) Trusts in April 1991 with a second wave in April 1992. Whether the formation of NHS Trusts is regarded as an opportunity or a threat, it is estimated that most NHS hospitals in the UK will become Trusts within the next 5 years. Bradford hospitals were among the first to become a Trust and this article discusses the experience to date and offers hints as to how other departments might prepare for Trust status. PMID- 8409228 TI - A Master of Science course at the Cardiff School of Medical Photography. AB - Formal teaching and training in medical photography at Cardiff started in 1969 when a School of Medical Photography was established, as part of the Department of Medical Illustration, at the Cardiff Royal Infirmary. In the early 1970s the school was transferred with the Medical Illustration Department to the newly built University Hospital of Wales, and housed in planned accommodation at what is now the Institute of Health Care Studies. The school offered courses in medical photography at the level of the qualifying examinations of the Professional Institutes, to suitably qualified in-service students appointed to the Medical Illustration Department as Trainee Medical Photographers. In 1990, the University of Wales approved a Master of Science course in Medical Illustration (Photography and Video) offered by the school. The course is available both to in-service students of the school and to practising medical photographers as mature students on a distance learning programme. Details of the new course and its delivery are given. PMID- 8409229 TI - Electronic photography: a new age of medical imaging? AB - This is a critical overview of present conceptions of the introduction of electronic photography in medicine. It is not a complete list of products, rather it is a description of how the requirements of the physician have influenced medical illustration in the past and will continue to do so in the future. Video systems are widely used in medicine. Besides the learning and teaching of effects of television, minimal invasive surgery (MIS) has become reality through endoscopy, rapidly accepted worldwide. Documentation of endoscopic procedures and their effects is becoming routine. Therefore, the conversion of complex optical information into binary units is a logical development to save space for storage. The reproduction, storage and transfer of detailed images is already realized by digital camera systems, photo CD, scanners and picture archiving and communicating system (PACS). Now electronic imaging in medicine has to be regarded as a matter of routine. The real impact of accelerated editing will be shown in the future. PMID- 8409230 TI - Image manipulation and the question of ethics. AB - In a world of changing technology, image manipulation has become easier to achieve than ever before in the history of illustration. The quality of computer generated images is such that it is difficult to distinguish them from a conventional photographic image. This article asks whether the industry can, and should, set controls to regulate itself before regulations are set for it by outside agencies. It discusses possible conflicts between ethical questions associated with image manipulation and the rights of image makers to produce creative images. Reference is made to historical incidents of image manipulation for propaganda; and technological advancements which make possible seamless montages of images using computer software programs and colour printers. PMID- 8409231 TI - Cleavage of 3-phenoxybenzoic acid by chicken microsomal preparations. AB - 3-Phenoxybenzoic acid, labelled at either the 3-phenoxy[U-ring C-14] or carbonyl C-14, produced the corresponding labelled transitory metabolite 4'-hydroxy-3 phenoxybenzoic acid when incubated with chicken liver microsomal preparation. The other radioactive reaction product identified from the metabolism of the carbonyl labelled 3-phenoxybenzoic acid was labelled 3-hydroxybenzoic acid, whereas metabolism of the [U-ring] labelled 3-phenoxybenzoic acid produced labelled hydroquinone in addition to 4'-hydroxy-3-phenoxybenzoic acid. Formation of both 4'-hydroxy-3-phenoxybenzoic acid, and its subsequent conversion to 3 hydroxybenzoic andhydroquinone were NADPH and oxygen dependent. The reaction was inhibited by carbon monoxide, but not by piperonyl butoxide or metyrapone. The inhibition by carbon monoxide suggests that the enzyme(s) involved in the cleavage is a form of P450. The enzyme(s) is a monooxygenase and has requirement for NADPH and oxygen. PMID- 8409232 TI - Microbial metabolism of fluazifop-butyl. AB - A microbial mixed culture able to grow on fluazifop-butyl and fluazifop was isolated. Fluazifop degradation by this microbial population was studied either when the herbicide was applied as the sole carbon source or in the presence of a second carbon source (sodium acetate or sodium propionate). The degradation rate was enhanced by sodium propionate. The degradation was found to be stereoselective. The S-enantiomer of fluazifop was degraded at a much higher rate than the R-enantiomer. Fluazifop disappearance was accompanied by formation of three metabolites which were identified by UV, IR, MS and NMR analyses. The metabolites were shown to be: 4-(5-trifluoromethyl-2-pyridyl)oxyphenol, 5 trifluoromethyl-2- hydroxypyridine and 2-(5-trifluoro-methyl pyridyl)hydroxy acetate. PMID- 8409233 TI - Enzyme immunoassay based survey of precipitation and surface water for the presence of atrazine, metolachlor and 2,4-D. AB - The suitability of enzyme immunoassay (EIA) as a method of analysis for 2,4-D, atrazine and metolachlor contamination in water samples was determined by comparing EIA results to gas chromatography (GC) results. The comparison of EIA and GC results yielded a correlation coefficient of 0.92, 0.98 and 0.92 for 2,4 D, atrazine and metolachlor, respectively. EIA was used to monitor seasonal trends in the concentrations of 2,4-D, atrazine and metolachlor in surface water and precipitation throughout the province of Ontario, Canada. 2,4-D was detected in excess of 4 micrograms/L in urban creeks during the period of application. Concentrations of 43 and 9 micrograms/L of atrazine and metolachlor, respectively, were detected during the field application period in surface water samples from the Kintore Creek watershed. The levels of 2,4-D, atrazine and metolachlor detected exceeded the Canadian Water Quality Guidelines for the protection of fresh water aquatic life. Concentrations as high as 445 and 322 ng/L of atrazine and metolachlor, respectively, were detected in precipitation samples collected from 17 locations in Ontario during the herbicide application period. The EIA was shown to be qualitatively and quantitatively comparable to GC analysis. PMID- 8409234 TI - In vivo and in vitro studies on the effect of larvin and cypermethrin on adenosine triphosphatase activity of male rats. AB - Liver is removed from albino rat and assayed for total adenosine triphosphatase (ATPase) activity following administration with single and repeated doses of larvin and cypermethrin. The data indicate that the total (Na+, K+; Mg2+) dependent ATPase in the liver tissue is significantly inhibited by single and repeated doses of both insecticides. This inhibition is more pronounced by the repeated dose of cypermethrin than that of larvin. The in vitro study revealed that the inhibition encountered by different concentration of both larvin and cypermethrin is of the irreversible non-competitive type. This data indicate that these insecticides can cause biochemical and histopathological changes in the liver ATPase activity which may inhibit several biochemical functions of ATPase system such as: the active transport of metal ions, oxidative phosphorylation of liver cells and generally the muscle contraction. PMID- 8409235 TI - Forecasts of active life expectancy: policy and fiscal implications. AB - Age-related changes in functional status can be summarized by active life expectancy (ALE) measures. ALE is useful in assessing efforts to improve function and in determining a population's service needs. ALE disaggregates total life expectancy (TLE) into components representing degree and type of impairment. We illustrate the calculation of two ALE measures and their relations to health inputs and service use. First, scores are calculated from 27 measures of function for persons 65 and over, as reported in the National Long Term Care Survey (NLTCS). The scores are then used to calculate the two ALE measures. Results are compared to ALE calculated from the 1982, 1984, and 1989 NLTCS. PMID- 8409236 TI - The implications of assessment. PMID- 8409237 TI - Physical disability in older Americans. PMID- 8409238 TI - Fear of falling and low self-efficacy: a case of dependence in elderly persons. PMID- 8409239 TI - Psychosocial and behavioral dimensions of physical frailty. AB - Findings from a decade of research on physical frailty in diverse elderly populations are summarized and integrated. While much of the literature focuses on the physical and functional consequences of chronic and acute illness, we emphasize the psychosocial and behavioral aspects of physical frailty. The underlying assumption of our approach is that the psychological representation of illness and disability, as well as the social contexts in which they occur, are important determinants of their impact on patients and family members. We focus on both patients and their primary support persons and on the comparative analysis of different disabling conditions. A patient and family caregiver impact model is used to summarize major findings of our research program and to make recommendations for intervention research. PMID- 8409240 TI - A human factors analysis of ADL activities: a capability-demand approach. AB - Older adults frequently encounter difficulties performing daily living activities. Often times these difficulties arise because environmental demands create barriers which hinder task performance. Currently, there is little empirical data that relate environmental demands to functional capabilities of older adults. The concepts and methods of Human Factors Engineering can be used to accomplish this goal. Human Factors views task performance within a systems context and maintains that successful task performance is dependent on a match between task demands and human capabilities. This article will discuss how Human Factors methodologies can be used to analyze problems encountered by older adults performing routine activities. Data from a study concerned with identifying physiological demands associated with personal and instrumental activities of daily living will be used to demonstrate the utility of using this approach. PMID- 8409241 TI - Hormonal regulation of motor behavior in senescence. PMID- 8409242 TI - Age-related changes in posture and movement. PMID- 8409243 TI - Cardiovascular, neuromuscular, and metabolic alterations with age leading to frailty. AB - As members of our society live longer, a greater percentage of the population will be older. These demographic changes will stress our social and medical delivery system, unless interventions can alter the course leading to frailty. Maximal aerobic power decreases with age, due to a decrease in cardiac output, and is exacerbated by cardiovascular disease. Asymptomatic aging does not reduce cardiovascular function to an extent that would lead to loss of function. Metabolism, endurance, and contraction velocity and muscle strength remain relatively high until 40, 50, and 60 years of age, respectively. After age 60, there are dramatic decreases (approximately 10% per year) which lead to loss of function and independence. The loss of muscle function leads to an increase in the likelihood of falls (approximately 4-fold). Exercise programs utilizing "aerobic" exercise activities do not lead to an increase in muscle function, whereas programs designed specifically for muscle can increase function and, presumably, reduce the risk of falls and injuries. PMID- 8409244 TI - Altered sensory function and balance in older persons. AB - The effects of three different visual inputs (eyes open, eyes closed, and inaccurate) while standing on an unstable surface (respectively, UEO, UEC, UI) were compared in a group of healthy elderly community dwellers (N = 239; mean age 76) and young (N = 34; mean age 34) adults. Subjects with medical factors known to affect balance were excluded. Body sway and loss of balance measured dynamic force platform (EquiTest, NeuroCom International). Visual and somatosensory inputs were rendered inaccurate through tilting of the standing surface and/or the visual surround proportional to the subject's angle of sway (sway referencing). The elderly group had significantly more difficulty balancing during UEC and UI, and a larger proportion lost their balance during UI. These findings are compared to those of other dynamic posturography studies in which sensory inputs were controlled. Issues of age, frailty, screening, and test protocol are addressed in order to account for differences in results among studies. PMID- 8409245 TI - The etiology and reversibility of muscle dysfunction in the aged. AB - Muscle weakness in the elderly is prevalent and morbid, closely linked to the frailty, functional decline, immobility, falls, and injuries in this population. The marked decrease in skeletal muscle strength and size with aging is a multifactorial syndrome which may be attributable in part to: (a) biological changes of aging itself; (b) the accumulation of acute and chronic diseases; (c) the assumption of a sedentary life style, and (d) selective or generalized nutritional inadequacies. Inactivity and undernutrition are potentially at least partially reversible with appropriate interventions, and therefore the delineation of the attributable risk of these two factors to the muscle weakness of aging is a critical research goal. Similarly, identification of appropriate modalities of physical activity and nutrition which have positive effects on muscle physiology in the aged is the focus of major investigations currently. PMID- 8409246 TI - A medical-social intervention in a 70-year-old Swedish population: is it possible to postpone functional decline in aging? PMID- 8409247 TI - Marriage, spousal caregiving, and a husband's move to a nursing home. A changing role for the wife? AB - 1. Gerontological nurses, when working with elderly couples experiencing role transitions upon nursing home placement, must attend to the needs of the healthier spouse, as well as the institutionalized patient. 2. Letting the community-dwelling wife set the tone for what her continued role will be allows her to define her marriage in a way that serves her needs at this transitory time. 3. If a woman defines herself as a wife, the nurse or nurse clinician's task may be to aid her in redefining the role's tasks, responsibilities, and limitations imposed by the nursing home setting. 4. If a woman does not perceive herself as a wife, nurses must help her accept these feelings as natural, and support her need to establish an identity as an individual and not as part of a couple. PMID- 8409248 TI - Research considerations: stress and perceived health status in the rural elderly. AB - 1. Stress is recognized as a significant factor in the development of acute and chronic illnesses in adults. The rural aged experience high stress levels and poor-to-fair health. 2. Commonly reported stressful events and chronic strains in the rural elderly involve loss, isolation, decreased abilities to manage self care activities, and financial concerns. 3. The older, single subjects in this study reported more stress and poorer health than younger, married subjects. 4. Nurses must develop strategies specifically designed to mediate the stress experienced by the rural aged. PMID- 8409250 TI - Patient self-determination: is anyone really listening? PMID- 8409249 TI - Dementia care creating a therapeutic milieu. AB - 1. In dementia care, the full spectrum of interventions in a therapeutic milieu provides for safety, structure, support, involvement, and validation. 2. A safe environment accommodates wandering and compensates for physical and cognitive impairments. A therapeutic milieu provides for both physical and psychologic safety. 3. Two components of structure include the design of the physical environment and the schedule of activities. 4. The central element that defines quality in a therapeutic milieu is the dimension of interpersonal relationships. PMID- 8409251 TI - Older women: social policies and health care. PMID- 8409252 TI - Developmental process: family caregivers of demented Japanese. AB - 1. Developmental process of caring for demented elders: the case of family caregivers consists of the beginning process (stage 1 through 4) and the awakening process (stage 5 through 7). 2. In the beginning process, the caregiver is unaware of the interaction between him/her and the demented family member, relies on conventional, general-purpose methods of care and on verbal communication, and consider care in terms of training and discipline of the person cared for. 3. The caregiver in the awakening process has realized that there is interaction between him/her and the demented family member, reflects on the nonverbal signs shown by the elder in order to improve their care, and provide warm and considerate care by making the most of nonverbal communication. PMID- 8409253 TI - Nursing home residents functional ability and perceptions of choice. AB - 1. Nursing home residents have the right to be involved in decision making relative to their care, which researchers have demonstrated has a positive effect on residents. 2. The authors designed a study to explore discrepancies between residents' perceptions of choice and the amount of choice preferred and their relationship to self-care abilities and functional abilities in nursing home residents. 3. The results indicate that residents desired more choice than they felt they were given, but that this difference was not related to their perceptions of their self-care abilities. As residents' functional abilities increased, actual choice as well as desired choice increased slightly. 4. Nurses should structure the nursing home's approach to residents so that choice is maximized within the constraints of the institution. PMID- 8409254 TI - A comparative study: guardianship petitions for adults and elder adults. AB - 1. The appointment of a guardian is the result of a court proceeding that transfers to another an individual's authority to make personal decisions. 2. Gerontological nurses may be asked to assist clients and families in the resolution of problems concerning personal and health care treatment decisions. 3. This study showed that adults younger than 65 years old needed a guardian due to mental retardation, mental illness, or accidental injury. Adults over 65 years old needed a guardian because they had mental retardation, alcoholism, dementia, or other physical disabilities. 4. Immediate family members were most often the individuals to serve as guardians. PMID- 8409255 TI - Geriatric falls: prevention strategies for the staff. AB - 1. Multiple falls and injuries are more prevalent among elderly over the age of 75 and are the second leading cause of accidental death in the elderly. The risk for falling is noted to be significantly greater in the hospitalized elderly. 2. Review of retrospective quality improvement chart audits revealed that peak fall times were associated with the patient's need for toileting, rest, and obtaining nutrition and hydration. 3. The MetroHealth Falls Prevention Program is based on simple proactive measures to prevent falls in the elderly. 4. An effective falls prevention program has several implications for gerontological nursing practice, including less restraint use, increased patient autonomy, and decreased loss of self-esteem. There is also a sense of increased nursing control over patient safety and time management, as well as implications for further nursing research. PMID- 8409256 TI - Gerontological research. Evaluating the process. AB - 1. Gerontological research is a nucleus for scientific investigations. Federal funding priorities include long-term care, iatrogenic complications, quality and continuity of care, self-care, and coping/adaptation of patients and caregivers. 2. Replication, meta-analysis, and synthesis of existing data are potential strategies for building on previous research. 3. Although the quantity of nursing research is limited, findings from nursing and allied health studies provide useful contributions to nursing practice. PMID- 8409257 TI - Mental health care reform and gerontological nursing. PMID- 8409258 TI - Dry mouth in the elderly. PMID- 8409259 TI - Women's strategies for living in a nursing home. AB - 1. Elderly women who appear to be managing their own lives even as nursing home residents were found in this study to employ 10 specific strategies. 2. The study showed that the women wanted as much control as possible over their lives and their day-to-day decisions. 3. Remaining active and, in particular, engaging in activities that encouraged them to remain mentally alert was important to them. 4. Sociability, including interaction with family members on a regular basis, was a strategy used by all participants. PMID- 8409260 TI - Cerebrovascular disorders of childhood. PMID- 8409261 TI - Etiology of stroke in children. AB - Cerebrovascular disorders are more common than once suspected, and our ability to diagnose stroke in children has improved with the development of newer imaging techniques in recent years. Children have a wide array of risk factors that promote cerebral infarction or hemorrhage, and a likely cause can eventually be pinpointed in about two thirds of patients if a thorough diagnostic evaluation is performed. Ideally, a systematic evaluation should confirm the presence of a cerebrovascular lesion and also identify the cause, concentrating initially on the more common or treatable risk factors. Recognition of the cause of a child's stroke is important, because the likelihood of recurrence depends largely on the etiology and whether treatment is available. PMID- 8409262 TI - Alternating hemiplegia syndrome: electroencephalogram, brain mapping, and brain perfusion SPECT scan study in a Chinese girl. AB - A 3-year-old Chinese girl with alternating hemiplegia syndrome failed to respond to anticonvulsants, antimigrainous drugs, and calcium channel blockers. She made a complete remission with a 4-week course of steroid, and relapsed after steroid withdrawal. Electroencephalogram and brain mapping during the hemiplegic attack showed unilateral high-voltage sharp slow-wave discharges in the temporo occipital region contralateral to the hemiplegic side and diffuse high-voltage slowing during attacks of quadriplegia or other clinical manifestation such as dullness, lethargy, or yawning. Brain perfusion single photon emission computed tomographic (SPECT) scan study during the attack showed decreased uptake in the temporoparietal region contralateral to the hemiplegic side and in the ipsilateral basal ganglia, whereas the perfusion was normal between attacks. Electroencephalogram background activity was improved while the child was in clinical remission with steroid treatment. Computed tomographic and magnetic resonance imaging scans of the brain were normal. Carotid angiogram failed to show any structural or dynamic changes of the carotid arteries. The possible mechanism underlying alternating hemiplegia syndrome might be transient and reversible cerebral ischemia with high-voltage slow-wave discharges shown in the electroencephalogram and decreased perfusion in SPECT scan. PMID- 8409263 TI - Technetium 99mTc-HMPAO SPECT in children and adolescents with neurologic disorders. AB - We evaluated regional cerebral blood flow with technetium 99mTc hexamethylpropyleneamineoxime single photon emission computed tomography (SPECT) in 20 children and adolescents with neurologic dysfunction of varied etiology and abnormal electroencephalograms (EEGs). All patients were also examined with computed tomography (CT) and magnetic resonance imaging (MRI). Abnormal perfusion was found in 17 (85%) of 20 SPECT scans. Abnormal CT or MRI scans were noted in nine (45%) and in 10 (50%) of 20 cases, respectively. In eight (73%) of 11 cases with normal CT scans and in seven (70%) of 10 with normal MRI scans, the SPECT scan was abnormal. Abnormal regional cerebral blood flow on SPECT scans correlated better with EEG abnormalities than with neurologic examination or CT or MRI scan findings. We conclude that in children and adolescents with a spectrum of neurologic diseases and abnormal EEGs, abnormalities of brain structure or function are more likely to be documented by SPECT than by CT or MRI scans. SPECT findings correlate well with the location and type of EEG abnormality. PMID- 8409264 TI - Chronic paroxysmal hemicrania in a young child: possible relation to ipsilateral occipital infarction. AB - We report a child with chronic paroxysmal hemicrania beginning at 3 years of age with more than 2 years of daily episodes. Indomethacin produced total relief. An ipsilateral, occipital hemorrhagic infarction, probably predating the headaches, may have contributed to their pathogenesis. This is an unusually early onset and persistent chronic paroxysmal hemicrania of possible symptomatic type. PMID- 8409265 TI - Responses of cerebral blood volume and oxygenation to carotid ligation and hypoxia in young rabbits: near-infrared spectroscopy study. AB - After ligation of the bilateral common carotid arteries with exposure to hypoxia in young rabbits, cerebral blood oxyhemoglobin, deoxyhemoglobin, and total hemoglobin were measured by near-infrared spectroscopy. Ligation of common carotid arteries decreased oxyhemoglobin, increased deoxyhemoglobin, and slightly decreased total hemoglobin. Exposure to hypoxia with bilateral carotid ligation more remarkably decreased oxyhemoglobin and increased deoxyhemoglobin than that without carotid ligation. However, the total cerebral blood volume did not change very much. This marked reduction in the cerebral oxygenation may cause brain damage and may be an important monitoring marker for the prevention of hypoxic ischemic encephalopathy. PMID- 8409266 TI - Proton magnetic resonance imaging and spectroscopic studies of the pathogenesis and treatment of juvenile dermatomyositis. AB - Juvenile dermatomyositis is an inflammatory disorder of muscle, skin, and connective tissue. Immune vasculopathy is central to the pathophysiology. We studied a 13-year-old boy with juvenile dermatomyositis using proton magnetic resonance imaging (MRI) with short tau inversion recovery (STIR), and proton magnetic resonance spectroscopy (MRS) to quantitate lipid and water in affected regions of muscle. Tissue perfusion was assessed by measuring tissue water concentration changes during isometric exercise of the tibialis anterior muscle. During sequential studies over 3 months of steroid treatment, STIR image abnormalities, resting water concentrations, and diminished perfusion returned to normal. Resting lipid concentrations increased during this period. MRI serves to guide muscle biopsy and monitor progress of the disease state. MRS demonstrates the vasculopathy and provides noninvasive assessment of steroid therapy in juvenile dermatomyositis. PMID- 8409267 TI - Stroke in children within a major metropolitan area: the surprising importance of intracerebral hemorrhage. AB - Our objective was to determine the incidence rate of stroke and stroke subtypes in children. We reviewed the medical records, autopsy records, and brain imaging studies of all children with a possible stroke within the Greater Cincinnati metropolitan area population of nearly 1.3 million during 1988 and 1989. Traumatic brain hemorrhages and germinal matrix hemorrhages were excluded. Of the 295,577 children in Greater Cincinnati, medical records of 178 children were screened. Sixteen cases (13 whites and three blacks) less than age 15 years fit strictly defined criteria for first-ever stroke. The incidence rate for cerebral infarction was 1.2 cases per 100,000 (95% confidence interval, 0.3 to 2.0). The combined incidence rate for intracerebral hemorrhage and subarachnoid hemorrhage was 1.5 cases per 100,000 children (95% confidence interval, 0.4 to 2.3). The incidence rate of all stroke in white children was 2.6 cases per 100,000 (95% confidence interval, 1.2 to 4.1), compared to 3.1 per 100,000 in black children (95% confidence interval, 0 to 6.6). The combined 30-day mortality for intracerebral hemorrhage and subarachnoid hemorrhage was 22% (two of nine) compared to 14% (one of seven) for cerebral infarction. We conclude that in contrast to the picture in adults, hemorrhagic stroke among infants and children is at least as common as ischemic infarction. PMID- 8409268 TI - Stroke and mixed connective tissue disease. AB - We describe the clinical presentation and course of two girls with cerebrovascular disease and mixed connective tissue disease. One developed rapid onset hemiparesis and aphasia secondary to left internal carotid artery occlusion. She experienced a complete recovery after treatment with prednisone and cyclophosphamide. The other patient was diagnosed as having mixed connective tissue disease but had acute neurologic deterioration. She died due to an intracerebral hemorrhage. Autopsy demonstrated small-vessel fibrinoid necrosis. Although cerebrovascular disease secondary to central nervous system vasculitis is a manifestation of systemic lupus erythematosus, this is the first description of cerebrovascular disease as a primary sign in mixed connective tissue disease. These cases demonstrate the range of cerebrovascular disease observed in children with mixed connective tissue disease. PMID- 8409269 TI - Mobius syndrome: evidence for a vascular etiology. AB - We report five infants with restricted lateral gaze, facial diplegia, feeding difficulty, and/or respiratory disorders without significant pulmonary disease. Viral studies were negative in all patients. Two children had radiologic findings that included brain-stem hypoplasia and symmetric calcification in the dorsal tectum at the junction of the midbrain and pons. Autopsy of one of these two children demonstrated capillary telangiectasia in the mesencephalon and pons. The other three children had normal computed tomographic (CT) scans. However, their autopsies revealed focal brain-stem necrosis with calcifications but without vascular malformation. We suggest that the capillary malformations in one of our patients directly resulted in a vascular-induced necrosis and the manifestation of Mobius sequence. The similarity of symmetric neuropathologic findings in the three other patients and the CT scan in the one surviving patient suggest focal hemodynamic changes restricted to the posterior circulation, indirectly supporting a vascular theory of embryopathogenesis. PMID- 8409270 TI - Ischemic stroke in the young: evaluation and age comparison of patients six months to thirty-nine years. AB - Ischemic stroke in the young is uncommon, but we currently evaluate at least one young stroke patient at our institutions each week. We undertook this chart review of strokes in patients between the ages of 6 months and 39 years to review all conditions associated with, and thus possibly contributory to, the stroke. We also compare younger and older age groups to observe if age-dependent factors exist. Of 100 total ischemic strokes, 22 were in persons 6 months to 18 years and 78 were in persons 19 to 39 years. Seventy-five percent of strokes were associated with a condition known or postulated to increase stroke risk. Some of these conditions are well accepted as causes for stroke, such as some forms of heart disease, whereas others are only postulated, such as the hypercoagulable states. Taken as a whole, associated conditions were approximately equally divided between infectious/inflammatory, structural, and presumed hypercoagulable conditions. Strokes in the first two decades of life were more commonly associated with infectious/inflammatory conditions, whereas strokes in the next two decades more commonly had structural or presumed hypercoagulable associated conditions. Since many strokes remain unexplained, it would be valuable to determine the significance, if any, of conditions less well known as risk factors for stroke. PMID- 8409271 TI - Unique hereditary sensory and autonomic neuropathy with growth hormone deficiency. AB - The proband, a French-Canadian white boy, presented with congenital sensory polyneuropathy, moderate to severe sensorineural hearing loss, infantile cataracts, nystagmus, esotropia, unusual facies, hypotonia, bilateral congenital hip dysplasia, delayed ossification of the femoral heads, scoliosis, short stature secondary to growth hormone deficiency, and developmental delay. His parents are consanguineous. His maternal first cousin, a 16-year-old girl, has congenital sensory polyneuropathy, infantile cataracts, unusual facies, scoliosis, short stature secondary to growth hormone deficiency, late-childhood onset arthritis, and hypoglycemia. Reportedly, she has no hearing difficulties and has normal intelligence. Her parents are third cousins. These children appear to have a distinct variant of hereditary sensory and autonomic neuropathy with infantile cataracts, unusual facies, skeletal dysplasia, short stature secondary to growth hormone deficiency, and other features, with probable autosomal recessive inheritance. PMID- 8409272 TI - An alternative pathogenic mechanism for stroke in children: a role for the sympathetic nervous system. PMID- 8409273 TI - Ontogeny of a circadian rhythm in body temperature in newborn lambs reared independently of maternal time cues. AB - To investigate the ontogeny of a circadian rhythm in body temperature, we performed 24-h temperature measurements at six postnatal ages in ten newborn lambs reared independently of their ewes. At 18 to 48 h of age, a significant time-of-day variation in body temperature was found, the variation achieving a peak between 5.00 pm and 6.00 pm. Over the next two weeks, this time-of-day variation disappeared, reappearing at 18 days of age, although the phase had shifted such that the peak temperature was achieved between 8.00 am and 9.00 am. At 26 days of age, the time-of-day variation in body temperature was still present, maintaining the same phase relationship as at 18 days of age. It is possible that the significant time-of-day variation observed in the two oldest groups is due to the emergence of a discernable circadian rhythm in body temperature entrained to the light-dark cycle during the second or third week of life, whereas the time-of-day variation observed in the 18 to 48-h old is due to residual effects of entrainment to maternal zeitgebers. We speculate that the lack of any observable time-of-day variation in the 4, 6 and 11 days-old lambs may denote the presence of no rhythm or possibly a free-running rhythm in body temperature in subjects not yet able to entrain to photic zeitgebers. PMID- 8409274 TI - Thyroxine and fetal blood coagulation: a fetal lamb study. AB - Glucocorticoids have been shown to accelerate the development of blood coagulation in the fetal lamb (Kisker, Robillard & Bohlken, 1983). The current studies examine the influence of high levels of triiodothyronine (T3) on the development of blood coagulation in the fetal lamb. Eight twin fetal lambs were studied during the last trimester of pregnancy (109-138 days gestation) using chronically placed arterial and venous catheters for infusion of T3 and withdrawal of blood samples. One fetus of each twin was infused intravenously with T3 at a constant rate of 0.6 micrograms T3/0.4 ml/h for 48 h. The other twin was infused with 5% dextrose at 0.4 ml/h for 48 h. Blood samples for measuring PT, PTT, TT, fibrinogen, factors II, V, VII, VIII, IX, X, XI and XII were obtained prior to and at the completion of the infusions. The results were analyzed for differences between the samples from paired control and T3 treated animals. Factor V significantly decreased in the T3 infused animals from 51.2% +/ 12.5% to 44.8% +/- 13.8% (P = 0.038). Factor VII also decreased in the treated animals 57.8% +/- 20% to 43.6% +/- 10.8%, but the changes were not statistically significant (P = 0.06). Factor XII significantly increased in the treated animals from 37% +/- 10.5% to 45% +/- 13.7% (P = 0.004). High levels of T3 in third trimester fetal lambs are accompanied by a moderate decrease in factor V and an increase in factor XII activity. PMID- 8409275 TI - Phospholipase A2 and arachidonic acid release from permeabilised myometrial cells from guinea pig uterus. AB - The release of arachidonic acid and inositol polyphosphates from permeabilised myocytes derived from guinea pig uterus has been studied. Both are enhanced by free calcium at 100 nM and 10 microM and particularly by 50 microM GTP gamma S. To distinguish between the contributions of phospholipase C and A2 to the release of arachidonic acid the phospholipase C inhibitor neomycin was used. At 1 and 10 mM, but not at 0.1 mM, neomycin caused effective inhibition of inositol polyphosphate release of over 95%. Neomycin (1 mM) also reversed GTP gamma S stimulated, but not calcium-stimulated release of arachidonic acid. This action was reflected in changes in [3H]arachidonic acid labelling of the membrane phosphatidylinositol and phosphatidylcholine pools, which were depressed by over 20% on the addition of 50 microM GTP gamma S, an effect completely reversed by 1 mM neomycin. The effects of neomycin were much more pronounced on inositol phosphate than on arachidonic acid release. The ability of 1 mM neomycin to inhibit arachidonic acid release was reversed by addition of 1 microM phorbol 12 myristate 13-acetate, implying a role for protein kinase C activation in stimulation of arachidonic acid release. Measurement of phospholipase A2 activity with 1-stearoyl 2-arachidonoyl phosphatidylcholine as exogenous substrate demonstrated the ability of 1 and 10 mM neomycin to inhibit the enzyme particularly when it was maximally activated with 1 mM free calcium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409276 TI - Fetal and neonatal action of a polycyclic hydrocarbon (benzpyrene) or a synthetic steroid hormone (allylestrenol) as reflected by the sexual behaviour of adult rats. AB - Allylestrenol or benzpyrene, given either between the 15th and 19th days of fetal life or from birth to the postnatal 7th day, caused dramatic decrease in the sexual activity of adult female rats. Allylestrenol, given in fetal life, resulted in a profound increase in the sexual activity of male rats. Given in newborn conditions, a decrease of sexual activity was caused by the same chemical. Considering, that one of the molecules has importance in medical practice and the other is an environmental pollutant, the experiments forecast the probability of modification in the sexual behaviour of human adults. PMID- 8409277 TI - Influence of sleep on systemic and coronary hemodynamics in lambs. AB - Experiments were done on seven lambs between the ages of 18 and 24 days to investigate the effects of sleep on systemic and coronary hemodynamics. Each lamb was anesthetized and instrumented for recordings of electrocorticogram, electro oculogram and nuchal electromyograms, and for measurements of cardiac output and coronary blood flow as well as systemic arterial blood pressure and arterial-, mixed-venous- and coronary-sinus- hemoglobin oxygen saturations. No sooner than three days after surgery, measurements were made during periods of quiet wakefulness (QW), quiet sleep (QS) and active sleep (AS) at an ambient temperature of 25 degrees C. Cardiac output and heart rate were decreased during AS compared to QW and QS. A significant increase in systemic vascular resistance during AS prevented more than a slight decrease in systemic arterial blood pressure. Coronary blood flow decreased during AS compared to QW and QS. Furthermore, myocardial work as estimated from the product of systolic blood pressure and heart rate decreased during AS compared to QW and QS as did myocardial oxygen consumption and myocardial oxygen transport. Transient hypertensive phases--defined as transient increases in mean blood pressure of greater than 15 mmHg above the mean blood pressure for a period of AS--occurred during eight of 13 periods of AS in four of the seven animals. These transient hypertensive phases were caused primarily by an increase in systemic vascular resistance; surprisingly, coronary vascular resistance also increased at this time when estimated myocardial work was increased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409278 TI - Cerebral oxidative metabolism in fetal sheep with prolonged and graded hypoxemia. AB - Cerebral oxidative metabolism and associated circulatory responses were determined in 14 unanesthetized fetal sheep near term, during a normoxic control period and subsequently, during four days of prolonged and graded hypoxemia induced by progressively lowering maternal inspired oxygen concentration with 1 2% CO2 added; first day 18%, second day 16%, third day 12-14%, fourth day 10-12%. Preductal arterial and sagittal vein blood samples were analyzed for oxygen content, blood gas tensions and pH. Regional blood flow was measured with a microsphere technique. Cerebral blood flow increased in a stepwise manner with the graded reduction in fetal arterial O2 saturation and continued to be well predicted by blood gas and metabolic alterations, with no adaptive change evident. Cerebral oxidative metabolism remained little changed with chronically induced hypoxemia until arterial O2 saturation was less than 30% and with fetal acidemia evident when decreased to 70% of normoxic control values. Whether the decrease in oxidative metabolism by the brain at this time represents an adaptive response whereby growth and functional alterations lead to a decrease in nonessential energy utilization or rather a pathological change, remains to be determined. PMID- 8409279 TI - Cardiorespiratory costs of growth in low birth weight infants. AB - The energy cost of growth includes two components: the energy stored in new tissues and the energy expended in all energy requiring steps associated with nutrient intake and net tissue accretion. Most of the energy expended in growth is accounted for by the energy cost of tissue anabolism: peptide bonds, lipogenesis, substrate transport, etc. However, to the extent that additional work is required of the heart and lungs for growth-related increases in O2 and CO2 transport, increased energy is also expended in cardiorespiratory work. Indirect estimates of these costs can be gained by examining the effects of diet and weight gain on heart rate and respiratory frequency. We studied 66 healthy low birth weight infants, mean study weight = 2010 g, fed constant intakes of protein (2.25-3.9 g/kg per day) and energy (100-150 kcal/kg per day). These diets led to rates of weight gain ranging from 13.9 to 21.7 g/kg per day, among the diet groups. Bi-weekly 6-h assessments of energy expenditure, heart rate, respiratory frequency and state of sleep were made after full enteral intake was achieved. After adjustment of heart rate for the effect of postnatal age, heart rate during active sleep was related to weight gain (y = 0.97 x + 144, r2 = 0.15), nitrogen-energy ratio of the diet (y = 5.9 x + 139,2 r2 = 0.22), and energy expenditure (y = 0.53 x + 129, r2 = 0.13). Multiple regression analysis revealed that age-adjusted heart rate during active and quiet sleep was significantly related to a combination of the same three variables (r2 = 0.31).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409280 TI - Epithelial cell invasiveness of non-enteropathogenic serotypes of Escherichia coli. AB - Current evidence suggests that enteropathogenic Escherichia coli (EPEC) of traditional serotypes possess a three-stage pathogenesis: localised adherence (LA), to, attachment-effacement (AE) of, and penetration of, enterocytes, all of which can be reproduced in tissue culture models in vitro. Three E. coli isolates of non-traditional serotypes (02:H2, 02:H25 and 015:H2) isolated from children with diarrhoea were previously shown to be positive for LA and AE activities in laboratory models. In the present study, they were, in addition, shown to be positive for invasion of a HEp-2 cell monolayer. These findings further establish the pathogenicity of non-traditional serotypes of E. coli and their role in the causation of diarrhoea. PMID- 8409282 TI - Bibliography on diarrhoeal diseases. PMID- 8409281 TI - Vibrio spp. isolated from natural waters of the city of Yangon, Myanmar. AB - Virulence properties of the environmental isolate of vibrios from natural waters of Yangon, Myanmar were studied. Vibrio spp. were isolated for identification by the membrane filtration method and cultured on thio-sulphate-bile-sucrose media. No Vibrio cholerae O1 were isolated. V. cholerae non-O1 were the major vibrio species isolated from the samples. None of them were detected for cholera-toxin like toxin, thermostable direct haemolysin, or heat-stable enterotoxin. Sixty-one isolates gave haemolytic activity; 27 V. cholerae non-O1 and 6 V. fluvialis isolates produced protease. PMID- 8409283 TI - New strains of Vibrio cholerae O139 in India and Bangladesh: lessons from the recent epidemics. PMID- 8409284 TI - In vitro uptake of amino acids in the jejunal mucosa of patients with cholera. AB - In vitro uptake of 14C-labelled amino acids was studied in jejunal mucosa biopsy specimens from 64 adults admitted for treatment of cholera (proven by stool culture) within 48 hours of onset of watery diarrhoea to determine the state of amino acid carriers in the jejunal mucosa during actively purging disease. Continued absorption of amino acids by the NBB carrier (for neutral amino acids), the Y+ system (for dibasic amino acids), and the PHE carrier were operative even during the actively purging stage of watery diarrhoea due to cholera. The IMINO carrier for absorption of N-substituted amino acids was found to be inoperative during cholera but the imino acids could be absorbed by the PHE carrier. This study demonstrates continued intestinal absorption of amino acids during cholera, provides scientific basis for use of amino acids in "improved" oral rehydration solutions utilising amino acid transport systems which are linked to the absorption of sodium (and water) so that reduction in diarrhoeal stools can be achieved, and emphasises the importance of maintaining feeding during acute diarrhoea to prevent the development of malnutrition. PMID- 8409285 TI - Serum and coproantibody responses to rotavirus infection in children during the first two years of life. AB - The serum and coproantibody response (IgG and IgA) to naturally acquired rotavirus infections and reinfections in the first two years of life were studied in 13 cases and 12 control children using enzyme-immunoassay. A 100% IgG seroconversion and coproconversion occurred after tertiary and quaternary infections respectively. Unlike the IgA coproantibody response, serum IgG was detected more frequently after symptomatic (89%) than asymptomatic (50%) infections, and its duration and titre were boosted by reinfections. The durations of both responses was prolonged when reinfection occurred at 15 months versus 8 months. Both responses were observed after primary infection in the first two months of life. Higher instances of virus reinfections were detected serologically (72.5%) than virologically (27%). The results suggest that at least 3 doses of a rotavirus vaccine will be required before 4 months plus boosters around the 10th and 15th months. Further, during vaccine trials, virus infections should be monitored virologically and serologically. PMID- 8409286 TI - Epidemiological study of infantile rotavirus diarrhoea in Tananarive (Madagascar). AB - An epidemiological study of rotavirus infections was conducted in Tananrive, Madagascar, from November 1988 to October 1990. Rotavirus antigen was detected by ELISA in faecal specimens of 183 of 1,659 children with acute diarrhoea (11%) and in 11 of 631 specimens from children without diarrhoea (1.7%). Rotaviral diarrhoeas were most frequently found in infants aged 6 to 18 months and occurred throughout the year with a definite peak during the first winter months. Analysis of the viral RNA by polyacrylamide gel electrophoresis permitted the characterisation of 170 and 194 strains identified. Nine different electropherotypes (A-I) and one mixed infection were observed. The "short" electropherotypes (A-E) were predominant and represented 140 strains (82.4%), and the "long" electropherotypes (F-I and M) represented 30 strains (17.6%). The "short" electropherotype A (cafb) was the most frequent in our environment (45.3% of cases) and was predominant during the first 14 months of the study. The "long" electropherotype F (bbea) appeared in July 1990 and was predominant during the last three months. Among these children with diarrhoea, the presence of rotavirus was significantly associated with vomiting, fever, and moderate to severe dehydration. However, no significant differences in the occurrence of these symptoms were found between the "short" and "long" electropherotypes. PMID- 8409287 TI - Production of mucinase and neuraminidase and binding of Shigella to intestinal mucin. AB - Production of mucinase and neuraminidase by Shigella spp. and their ability to bind to mucin was investigated. All four species of Shigella produced these enzymes. Virulent and avirulent pairs of Shigella did not differ in their ability to produce the enzymes after 18 h of growth. However, a significant difference in neuraminidase production was noted in Shigella dysenteriae type 1 and S. boydii (12-15) at 10 h growth. Avirulent strains of S. dysenteriae type 1, S. flexneri 2a, and S. boydii bound significantly more amounts of mucin than their virulent counterparts. PMID- 8409288 TI - Comparative effects of nicotinic acid and nicotinamide on cholera toxin-induced secretion in rabbit ileum. AB - Nicotinic acid reduces the cholera-toxin induced fluid secretion in experimental animals but its toxicity at high doses prevent its therapeutic use in patients suffering from cholera. This study aimed to determine whether nicotinamide, the non toxic amide derivative of nicotinic acid, is as effective as nicotinic acid in inhibiting cholera toxin induced intestinal secretion in vivo. Four intestinal loops, with their blood supply intact, were isolated in 30 rabbits and injected with either (i) 30 mM mannitol, (ii) 30 mM mannitol + 10 micrograms cholera toxin, (iii) 30 mM glucose, or (iv) 30 mM glucose + 10 micrograms cholera toxin. These rabbits were then randomly assigned to three groups receiving intraluminally either 100 mg/kg of nicotinic acid, 100 mg/kg of nicotinamide, or 10 ml/kg of Ringer solution. Measurement of intestinal fluid accumulation showed that nicotinic acid, but not nicotinamide, significantly reduced cholera toxin induced intestinal secretion. PMID- 8409289 TI - Continuously infused cyclosporine at low dose is sufficient to avoid emergency colectomy in acute attacks of ulcerative colitis without the need for high-dose steroids. AB - A dosage of 4 mg/kg/day intravenous cyclosporine as an adjunct to high-dosage hydrocortisone has been recommended to avoid colectomy in acute steroid-resistant ulcerative colitis. In treating eight such patients, four of whom presented with toxic megacolon, we immediately tapered the steroid dosage and infused a lower dosage of only 2 mg/kg cyclosporine for 15 days to fit a therapeutic range of 60 240 ng/ml, as previously designed for a kidney transplant program. Seven of the eight (87.5%), including three with megacolon, went into remission and started the chronic phase of treatment; the eighth patient underwent colectomy. Of the seven, one died on day 3 of the chronic phase because of pulmonary embolism while in clinical remission, and another discontinued treatment. The other five (62.5%) remain in remission on 6 mg/kg oral cyclosporine, or have already switched from cyclosporine to azathioprine. Two episodes of reversible nephrotoxicity appeared in the chronic phase only. These results emphasize the efficacy and safety of cyclosporine in acute ulcerative colitis, but there is still a need for further dose-response and drug association relationship studies. PMID- 8409290 TI - Are antibiotics useful in the management of nontoxic severe ulcerative colitis? AB - Severely ill patients with ulcerative colitis are usually treated with parenteral corticoids. Those with signs of systemic toxicity usually receive antibiotics as well. However, the role of antibiotics in patients without high fever, profound leukocytosis, peritoneal signs or megacolon is not clear. Since 1985, seven patients who were admitted to the hospital with severe ulcerative colitis but without signs of toxicity have received broad spectrum antibiotics. The patients ranged in age from 21 to 65 years: four were male; all were white. Stools for infectious agents were negative. Each patient received intravenous prednisolone at a dose of 60 mg/24 h. After 7 days, each patient continued to have eight to 10 bloody stools per day with low-grade fever and leukocytosis. Institution of broad spectrum antibiotics was associated with a striking resolution of bloody diarrhea and normalization of temperature. No patient has required colectomy in follow-up of 2-9 years. Whether the antibiotics are treating an undetected pathogen or eliminating non-pathogenic bacteria contributing to the inflammatory process is not clear. I suggest that severely ill patients with ulcerative colitis deserve a trial of antibiotics before being considered failures of medical therapy. PMID- 8409291 TI - Cluster of inflammatory bowel disease in three close college friends? AB - Although the etiology of inflammatory bowel disease remains unknown, current evidence favors the interplay of genetic predisposition with environmental factors. Clustering of inflammatory bowel disease in spouses had been described, and supports a role for an environmental agent. We describe three unrelated, unmarried men all attorneys who enjoyed a close, sustained friendship in college, and who within a decade of their contact developed inflammatory bowel disease. We describe the nature of their college contact, the course of their inflammatory bowel disease, and relate our cluster to the scanty literature on clustering in inflammatory bowel disease. Physicians should report similar observations to illuminate the roles of genetics and environment in the development of these illnesses. PMID- 8409292 TI - Dilation therapy for gastric outlet obstruction. Are balloons a bust? AB - Balloon dilation of gastric outlet obstruction has evolved over the past 10 years from a technical tour de force to a commonly applied treatment modality complementing or competing with conventional surgical therapy. While not risk free, current data suggest that 60-80% of patients undergoing such dilation therapy have sustained relief of their obstructive symptoms although long term acid suppression or cessation of nonsteroidal anti-inflammatory drugs may be necessary. PMID- 8409293 TI - A patient knowledge questionnaire in inflammatory bowel disease. AB - A multiple choice knowledge questionnaire was developed for patients with inflammatory bowel disease (IBD). Initially, 20 questionnaires were piloted, and then questions of poor discriminatory ability (> 75% patients giving a correct response) were eliminated. A second pilot study proved the internal consistency (coefficient alpha = 0.84) and the stability of the questionnaire using a test retest method (Spearman R = 0.86, p = 0.002). The questionnaire was then completed by 60 randomly selected patients attending colitis clinic. The mean score was 13 of a total of 36 (range 2-29). Eighty percent of patients wished to know more about their disease. Knowledge score was higher in patients who were members of the National Association of Crohn's Disease and Colitis (NACC) (p < 0.005) and in patients with Crohn's disease (CD) rather than ulcerative colitis (UC) (p < 0.005). Knowledge score correlated with the number of years spent in full-time education (R = 0.48, p < 0.001) and inversely with age (R = -0.33, p < 0.02) but did not correlate with disease duration. The study demonstrated some misunderstandings about IBD although most patients would have liked more information. The questionnaire could be used as a tool to evaluate patient education programs. PMID- 8409294 TI - Streptococcus bovis does not selectively colonize colorectal cancer and polyps. AB - The objective was to determine if Streptococcus bovis selectively colonizes colorectal cancer and polyps. Stools were submitted before colonoscopy; fluid and selected tissue biopsies obtained during colonoscopy were cultured for S. bovis. The setting was a large multispecialty clinic. Outpatients undergoing colonoscopy for suspected colorectal cancer and polyps were the participants. Forty studies on 35 patients were performed. One of 35 stools (2.9%) obtained before colonoscopy yielded S. bovis. S. bovis was cultured from three of 40 fluid aspirations (7.5%), one of 33 adenomas biopsied (3%), none of six adenocarcinomas, none of 14 nonneoplastic polyps, one of 40 (2.5%) biopsies from normal mucosa adjacent to an adenoma, and none of 40 mucosal biopsies remote from any lesion. These data do not support selective colonization of colorectal neoplasms by S. bovis. PMID- 8409295 TI - Liver biopsy in fever of unknown origin. A reappraisal. AB - We assessed the value of liver biopsy in the diagnosis of fever of unknown origin (FUO) in hospital-based patients by a retrospective analysis of all cases (24 cases) seen at the University of Michigan Medical Center over a 5-year period. Based on the findings of a liver biopsy performed in the course of the evaluation of FUO, patients were divided into two groups: a diagnostic group, in which an abnormal liver biopsy was helpful in determining the cause of the FUO, and a nondiagnostic group, which included those who had either normal biopsy results or abnormal biopsy results that did not lead to a final diagnosis. Four patients (16.7%) had diagnostic liver biopsy results (histoplasmosis in three and tuberculosis in one). Physical findings, such as hepatomegaly, and laboratory data, including routine liver chemistries, were not predictive of a diagnostic liver biopsy. Therefore, despite advances in diagnostic technology since this subject was last reviewed, liver biopsy continues to be useful in the diagnosis of FUO. Furthermore, in endemic or borderline endemic areas, histoplasmosis should be considered in the differential diagnosis of FUO, and liver biopsy can be helpful to confirm this diagnosis. PMID- 8409296 TI - Does exclusion of enteral lipid assist remission in Crohn's disease? AB - Based on the improvement of Crohn's disease (CD) with elemental diets, we treated a patient with recurrent stenosing disease using prednisolone and drastic reduction of oral lipid intake. Complete symptomatic and good radiological improvement took place. A newly proposed hypothesis for CD suggests that enteral lipids together with certain luminal bacterial fragments become strongly antigenic to produce an inflammatory reaction analogous to the adjuvant response. Because of the apparent success of dietary lipid exclusion in this patient, we suggest that it be considered as a future option for management of acute CD. PMID- 8409297 TI - High-volume postobstructive choleresis after transhepatic external biliary drainage resolves with conversion to internal drainage. AB - We report high-volume postobstructive choleresis in two patients who underwent transhepatic external drainage for malignant biliary obstruction. Excessive loss of bicarbonate-rich biliary fluid (up to 6.5 L/day) caused orthostatic hypotension, prerenal insufficiency, hyponatremia, and a decrease in serum bicarbonate. Therapy with isotonic fluids containing sodium, chloride, lactate, bicarbonate, and potassium was based on measurement of biliary fluid volume and electrolyte concentrations. Biliary fluid loss was terminated by conversion to internal biliary drainage. The reason for this rare complication of external drainage of biliary obstruction is unknown, but such patients must be closely monitored for volume loss. When high-volume choleresis occurs, biliary fluid and electrolyte losses should be precisely measured and replaced, and external biliary drainage converted to internal drainage. PMID- 8409298 TI - Injection therapy for colonic diverticular bleeding. A case study. AB - We report a successful colonoscopic identification of acute massive diverticular bleeding in an elderly patient and successful injection therapy with epinephrine. Colonoscopy is a valuable diagnostic tool in acute lower gastrointestinal (LGI) bleeding when the colon can be adequately visualized. Peridiverticular injection therapy should be considered a safe and effective therapeutic option when the bleeding site can be established. PMID- 8409299 TI - A unique presentation of lymphoma of the colon. AB - Non-Hodgkin's lymphoma often involves the gastrointestinal tract, but primary lymphoma of the colon is rare. Primary lymphoma of the colon masquerading as inflammatory bowel disease is even rarer. We report a patient with an initial diagnosis of Crohn's colitis, who later appeared to have had non-Hodgkin's lymphoma of the colon. We review the diagnosis and treatment of this rare disease. PMID- 8409300 TI - Effectiveness of cisapride in gastric ulcer. Results of a double-blind randomized trial versus ranitidine and versus cisapride plus ranitidine. AB - Among the factors involved in the pathogenesis of gastric ulcer, the reduced clearing capacity of the stomach seems to play an important role. On this basis, cisapride, which improves gastrointestinal motility, enhances gastric emptying, and prevents duodenogastric reflux, may be effective in the treatment of the gastric ulcer. We randomly allocated 60 consecutive patients, with uncomplicated antral gastric ulcer (diameter 5-25 mm), into three groups of treatment: cisapride 20 mg b.i.d. (C), ranitidine 150 mg b.i.d. (R), cisapride 20 mg b.i.d. + ranitidine 150 mg b.i.d. (C+R). Endoscopic examination with biopsy specimens was performed on admission, after 4 weeks and (if ulcer not healed) after 8 weeks of therapy. Three patients were lost to follow-up (two in C and one in C+R), and three were withdrawn, due to malignant ulcer (one case in R) or to side effects (one case of diarrhea in C, one case of headache in C+R). Healing rates at 4 weeks were 41.1% in C, 52.6% in R, and 50.0% in C+R; at 8 weeks they were 88.2% in C, 89.4% in R, and 94.4% in C+R. Though the lack of a placebo arm makes final considerations difficult, the results were similar in all three groups, with no evident differences. In conclusion, therapy with cisapride appears as effective as H2-blocker alone or combined treatments in healing benign gastric ulcer. PMID- 8409301 TI - Liver disease from surreptitious administration of urethane. AB - We report subacute necrosis, peliosis hepatis, venoocclusive disease and hepatic angiosarcoma after long-term administration of urethane. We take this to be the 12th case of urethane-induced hepatitis and the first associated with vascular liver tumor. PMID- 8409302 TI - Focal ultrasound lesions in cirrhotic liver diagnosed as regenerative nodules by biopsy. A morphometric analysis. AB - Ten cirrhotic patients with ultrasonically discernible focal liver masses underwent fine cutting needle biopsy. Specimens were obtained from the focal lesions under ultrasound guidance and histologically diagnosed as regenerative nodules. An image analyzer was then used to determine the cytoplasmic area, nuclear area, and nuclear/cytoplasmic ratio for 100 randomly selected cells from each specimen. Data were then compared with data for specimens of normal liver tissue and data from patients with alcoholic or posthepatic cirrhosis or well differentiated hepatocellular carcinoma (HCC). The morphometric parameters for the group of regenerative nodule specimens fell within an intermediate range between those for HCC and the nondysplastic samples, strongly suggesting a preneoplastic nature. Nine of the 10 regenerative lesions showed liver cell dysplasia, and 3 of these patients developed HCC during follow-up. Ultrasonically discernible focal masses in a cirrhotic liver should be considered preneoplastic, if not neoplastic lesions and treated aggressively to prevent their progression to outright malignancy. PMID- 8409303 TI - Magnetic resonance imaging in inflammatory bowel disease. AB - Magnetic resonance (MR) images were acquired in 28 consecutive inflammatory bowel disease (IBD) patients undergoing medical treatment. The protocol employed included i.v. gadopentatate dimeglumine, pre- and post-contrast breath-hold Fast Low Angle Shot (FLASH), and fat-suppressed spin echo imaging. The percent contrast enhancement (% CE) of the fat-suppressed images was compared with severity of inflammation based on endoscopic and/or surgical findings. The %CE of the contrast-enhanced images was 169% +/- 63 in cases of severe inflammation (n = 16), 97% +/- 38 in moderate inflammation (n = 8), and 49% +/- 26 in mild inflammation (n = 4). Significant correlations were found between the clinicopathologic findings and (a) % CE, r = 0.74 (p = 0.0001); (b) length of affected bowel segment, r = 0.49 (p = 0.007); and (c) bowel wall thickness, r = 0.42 (p = 0.02). In a subsequent comparison, %CE was correlated with length of affected bowel and bowel wall thickness. The best correlation was bowel wall thickness, r = 0.53 (p < 0.004). Good correlation was found between MR findings and pathology/histology findings in the determination of bowel wall thickness, length of diseased bowel, and severity of inflammation in 10 patients who underwent bowel resection. The results of this study show that MR images demonstrate the extent and severity of inflammatory changes in the GI tract, which correlate with endoscopic and histological findings. PMID- 8409304 TI - Glycogenic acanthosis of the esophagus and gastroesophageal reflux. AB - A nodular appearance of the esophageal mucosa, observed in 3.5% of 2,328 consecutive upper endoscopic examinations, most commonly appeared as multiple, uniformly sized, oval or round elevations usually < 1 cm, involving otherwise normal esophageal mucosa. Endoscopic biopsies in 35 consecutive patients followed prospectively demonstrated the nodules to represent glycogenic acanthosis--a combination of cellular hyperplasia and increased cellular glycogen. When studied by 24-h ambulatory esophageal pH monitoring, 83% of these patients had pathologic gastroesophageal acid reflux; mean percentage of time with pH < 4.0 was 37.3%. Antireflux therapy improved symptoms in all patients but failed to eradicate the lesions of glycogenic acanthosis. Although its etiology and pathogenesis still remain elusive, glycogenic acanthosis may be related to gastroesophageal reflux. PMID- 8409305 TI - Atrophic chronic gastritis in patients with epidermoid carcinoma of the esophagus. PMID- 8409306 TI - Unusual endoscopic appearance of collagenous colitis. PMID- 8409307 TI - Massive rectal bleeding from an ulceration in a jejunal diverticulum. PMID- 8409308 TI - Colonic polyps found on flexible sigmoidoscopy: a retrospective study. PMID- 8409309 TI - Ischemic hepatitis in cirrhosis: not so rare, not always lethal. PMID- 8409310 TI - Granulomatous hepatitis and fever of unknown origin: influence of AIDS epidemics. PMID- 8409312 TI - Is the aggressive management of peptic ulcer justified by the data? AB - Sophisticated and very effective means of diagnosing and treating peptic ulcers are now available. This entails considerable resource utilization in affluent western societies, all with resource limitations, which mean that the costs need to be balanced by the outcomes achieved--the benefits. Little attention has been paid to the risk to life and well-being posed by ulcer and the effect, if any, of the intervention on such risks. I present an analysis of the natural history of peptic ulcer including studies of the natural history of peptic ulcer including studies incorporating life-table analysis and a population comparison. The conclusions are clear. The possessors of a peptic ulcer have, in general, a normal life expectancy. A major cause of ulcer-related death in the past was postoperative. Symptoms and workloss have diminished after the introduction of H2 antagonists, but no evidence is yet available of any change in the rates of bleeding, perforation or death from ulcer related to modern methods of diagnosis and management. The risk of major complication is approximately 0.025/year over the next 10 years. The risk of death is greatest in elderly women with gastric ulcer and in those on corticosteroids. The data suggest to me that the costs of aggressive investigation and treatment of peptic ulcer need to be justified by the dangers averted. PMID- 8409311 TI - Are routine duodenal and antral biopsies useful in the management of "functional" dyspepsia? A diagnostic and therapeutic study. AB - Fifty-three patients with previously uninvestigated chronic dyspepsia symptoms in the absence of gastrointestinal or extra-gastrointestinal disease (functional dyspepsia) underwent antral and duodenal mucosal biopsies to detect the role of such samplings in the presence of normal endoscopic findings. Patients were enrolled in a randomized, placebo-controlled, double-blind trial, receiving either eradicating treatment (colloidal bismuth subcitrate plus metronidazole) or placebo if they had Helicobacter pylori-associated gastritis (20 patients), or cisapride or placebo if they had normal antral mucosa (28 cases). Unsuspected celiac sprue was found in one patient. Eradicating treatment ameliorated histological gastritis (p = 0.01). However, owing to great placebo efficacy, symptom remission rates following a 1-month wash-out period in both treatment groups were no higher than that in controls. Independent of the initial randomization, an extremely low symptom recurrence rate was observed during a drug-free follow-up study equivalent to the mean duration of symptoms before enrollment. We conclude that in functional dyspepsia, bulbar and antral biopsies are not useful in clinical management, equivalent symptom relief can be achieved in patients randomly assigned to both drugs and placebos, and such improvement can be long lasting in the absence of any maintenance treatment. We believe the prevalence of unsuspected villous atrophy and the therapeutic role of investigation-based reassurance deserve further assessment. PMID- 8409313 TI - The effect of acute emotional stress on gastric acid secretion in normal subjects and duodenal ulcer patients. AB - Emotional stress (ES) has been proposed as a possible factor in the pathogenesis of duodenal ulcer (DU) disease. Modern, well-controlled studies on the effect of ES on gastric acid secretion (GAS) in both normal healthy subjects and patients with inactive DU are lacking. Ten normal (N) men and 10 men with inactive DU were observed on 2 separate days. In random order, subjects either underwent dichotomous listening (DL) to induce stress or a control (non-DL) test. In addition to measuring GAS in 15-min periods, heart rate and blood pressure were measured every 7.5 min, and visual analog scale measures of emotion (relaxation, anxiety, anger, tension, and depression) were monitored. Subjects underwent 2 separate study days, 1 h of a basal period followed by 1 h of a DL session or 1 h of a basal period followed by 1 h of a non-DL control session; the order of the days was randomized. In both N and DU emotional stress by DL induced these parameters significantly: increased heart rate; raised systolic and diastolic blood pressures (p < 0.01); increased anxiety, anger, and tension (p < 0.03); and decreased relaxation (p < 0.01). The non-DL control test did not alter cardiovascular or emotion measures in either group. While ES did not alter GAS in N subjects, ES increased GAS when compared to the basal state (p < 0.02) and when compared to the control test (p = 0.07).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409314 TI - Serum IgA antiendomysium antibody titers as a marker of intestinal involvement and diet compliance in adult celiac sprue. AB - IgA-class antiendomysium antibodies (IgA-EmAs) are a very sensitive and specific serological marker of celiac sprue. Using an indirect immunofluorescent method, we evaluated the kinetics of the antibody titers both during a gluten-free diet and after gluten was added, comparing them with the intestinal histological pattern. The IgA-EmA titers were evaluated on sera from 91 untreated adults with biopsy-proven celiac sprue and, when positive, were reappraised after different durations of gluten-free diet. Antibody titers were also retested in eight adolescents who had voluntarily discontinued the diet while they were on a free diet. The IgA-EmAs were detectable in 85 of 91 (93.40%) patients but in none of the 438 controls (100% specificity). The antibody titers ranged from 1:5 to 1:2,000 or more and the intestinal histopathological pattern paralleled the antibody titers. After gluten withdrawal, IgA-EmA titers declined to zero in all patients complying with their diet. Modifications in gut histopathologic condition paralleled IgA-EmA kinetics, although seroconversion to negativity preceded mucosal recovery. After a gluten rechallenge, deterioration in gut histopathologic condition followed EmA reversion to positivity. Three negative IgA-EmA tests did not reflect a worsening in gut histopathologic condition after a gluten-containing diet, thus making the diagnosis of celiac sprue uncertain. PMID- 8409315 TI - Bowel patterns and anxiety. Demographic factors. AB - In a survey of bowel patterns and anxiety on 1264 health maintenance organization (HMO) members undergoing health assessment, I found (a) Stool frequency increased with age (p = 0.001), was greater in men than women (p < 0.00001), and was greater in whites than blacks (p = 0.07); (b) Fecal incontinence increased with age in women (p < 0.001) but was not age-related in men (p > 0.10); (c) Laxative use was greater in women than men at all ages (p < 0.01), and there was an age effect on use in women (p < 0.025) but not in men (p > 0.20); (d) Bowel pattern change and abdominal pain were frequently caused by stress, and both effects declined with age in each gender (p < 0.05); (e) More women than men at all ages reported stress effects (p < 0.001), and subjects who reported either stress effect scored higher on both parts of the State-Trait Anxiety Inventory (p < 0.00001) than other people. Bowel patterns and their relation to anxiety have demographic characteristics. PMID- 8409316 TI - Does ozone alleviate AIDS diarrhea? AB - Five patients with acquired immune deficiency syndrome (AIDS) or AIDS-related complex (ARC) and intractable diarrhea were treated with daily colonic insufflations of medical ozone (oxygen/ozone mixture) for 21-28 days. The daily dose of ozone (O3) ranged from 2.7 to 30 mg. Three of the four patients whose diarrhea was of unknown etiology experienced complete resolution, and one patient had marked improvement. The fifth patient, whose diarrhea was due to Cryptosporidium, experienced no change. No consistent change in the absolute number of helper (CD4) or suppressor (CD8) lymphocytes was detected, and no obvious changes were seen in the PO2 or the results of routine hematologic and blood chemistry studies. Patients had mild to moderate local discomfort during ozone administration early in the course of treatment, but no adverse systemic effects were observed. The results of this series suggest that medical ozone administered by rectal insufflation is simple, safe, and effective. Should this simple treatment be used routinely to treat chronic intractable ARC/AIDS diarrhea? PMID- 8409317 TI - The use of GoLYTELY and Dulcolax in combination in outpatient colonoscopy. AB - One hundred and sixty patients were enrolled in a double-blind, placebo controlled trial to evaluate the effectiveness of the combination of the polyethylene glycol electrolyte lavage solution GoLYTELY (Braintree Laboratories, Inc., Braintree, MA, U.S.A.) and the contact laxative bisacodyl (Dulcolax, Boehringer Ingelheim Pharmaceuticals, Ridgefield, CT, U.S.A.) in improving outpatient preparation for colonoscopy. One hundred and fourteen patients completed the study; 59 received GoLYTELY followed by three Dulcolax capsules and 55 received GoLYTELY followed by three placebo capsules. A statistically significant improvement in the group receiving Dulcolax was noted in overall quality of preparation (p = 0.015) and amount of liquid stool present during colonoscopy (p = 0.02). We noted no significant difference in the occurrence of adverse effects. No patient in either group suffered from symptomatic orthostatic hypotension. These results suggest that an improved colonoscopy preparation can be obtained by combining a standard GoLYTELY preparation with three 5-mg Dulcolax tablets, without sacrificing patient safety or comfort. PMID- 8409318 TI - Pathogenetic aspects of persimmon bezoars. A case-control retrospective study. AB - To prove that ingestion of unpeeled persimmon is necessary for the development of a persimmon phytobezoar, we interviewed 15 patients, in most of whom this condition developed after peptic ulcer surgery. We compared the study group with a control group of 15 patients who had undergone peptic ulcer surgery but did not have a bezoar. In contrast with the control group, most patients with bezoars had ingested unpeeled fruits (p < 0.01). While ingestion of persimmon carried a 9.8 fold risk of bezoar development, ingestion of the unpeeled fruit increased the risk of this complication 56 times over that of age- and sex-matched controls. We recommend that patients who have undergone ulcer surgery be warned particularly against eating unpeeled persimmons. PMID- 8409319 TI - Metallic expanding biliary stents in malignant obstruction. Cases with stent in stent. AB - We reviewed a total of 13 stent (Gianturco-Rosch biliary Z-stent) placements in 11 patients with biliary obstruction due to malignancy and report the cases treated with the stent-in-stent technique for treatment of stent occlusion due to tumor ingrowth. Causes of biliary obstruction included cholangiocarcinoma (four cases), hepatic hilar metastasis of gastrointestinal carcinoma (five cases), and pancreatic carcinoma (two cases). After transient percutaneous transhepatic biliary drainage (PTBD), the stents were successfully inserted by transhepatic route in all patients without any serious complication that needed further surgical intervention. Almost all patients were freed from the external PTBD tube about a week after stent placement and discharged from the hospital with improvement in quality of life as well as normal serum bilirubin levels. The technical advantages of stent placement include ease of insertion and the ability to drain both right and left biliary systems from a single transhepatic route by arranging the stents in a variety of configurations. Furthermore, it provides a second chance of stent placement when the previous stent has been occluded by tumor ingrowth. PMID- 8409320 TI - Fulminant hepatic failure with massive necrosis as a result of hepatitis A infection. AB - Fulminant hepatic failure as a result of hepatitis A is a rarely diagnosed complication entity in developed countries. With the advent of specific serologic markers for acute hepatitis A virus infection, the incidence and pathology of fulminant hepatitis A can be more clearly defined. We describe four patients (one adult, three children; two males and two females, ages 2 1/2-58 years) referred to our institution for orthotopic liver transplantation subsequent to fulminant hepatic failure following hepatitis A infection. All of these patients had a history of residence in or travel to hepatitis A endemic areas. Hepatitis A infection was documented by the presence of serum IgM against hepatitis A virus prior to transplantation. Infection with hepatitis B virus, cytomegalovirus, Epstein-Barr virus, and herpes simplex virus was excluded by clinical and specific serologic examinations. All patients presented with varying degrees of encephalopathy, progressing to coma. Coagulopathy in the form of prolonged prothrombin time and partial thromboplastin time was present in all patients. Peak liver parenchymal enzymes increased to greater than ten times the upper limit of the normal range. Total and direct bilirubin levels increased to > 20 and 10 mg/dl, respectively. Histologic evaluation of the explanted livers showed a spectrum of changes ranging from periportal hepatocellular necrosis with focal parenchymal collapse and prominent bile duct proliferation to massive necrosis with complete loss of hepatic architecture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409321 TI - Acute fatty liver of pregnancy after exposure to toluene. A case report. AB - We describe a patient in whom acute fatty liver of pregnancy developed after long term toluene exposure. Acute fatty liver of pregnancy was documented by a liver biopsy specimen stained with oil red O and observed under electron microscopy. Although the mother did well, she gave birth to a stillborn whose autopsy showed visceral congestion and placental infarction. This case raises the hypothesis of a possible relationship between toluene exposure and acute fatty liver of pregnancy. PMID- 8409322 TI - Thrombocytopenic purpura as a manifestation of acute hepatitis A. AB - Extrahepatic autoimmune manifestations are rare in patients with acute hepatitis A infection. We describe a 34-year-old man in whom severe autoimmune thrombocytopenic purpura developed as the manifestation of acute hepatitis A infection. Thrombocytopenic purpura is rarely described in association with hepatitis A, but to our knowledge has never been reported as a manifestation of acute hepatitis A. PMID- 8409323 TI - Hepatic dysfunction after repeated isoflurane administration. AB - Isoflurane, a halogenated volatile anesthetic, has not been associated with a distinct hepatic injury syndrome, as has halothane. Previous cases of suspected isoflurane-induced hepatotoxicity have been reported but questioned. We report the case of a patient without previous liver disease who developed repeated episodes of hepatitis after repeated exposures to isoflurane. Although no biopsy study was conducted, the temporal relationships illustrated in this case strengthen the argument for isoflurane-induced hepatotoxicity. PMID- 8409324 TI - Implications for the ligase chain reaction in gastroenterology. AB - The ligase chain reaction (LCR) is a new DNA detection method that uses thermostable ligase to discriminate exquisitely and amplify single base changes in genes of medical interest. This enzyme specifically links two adjacent oligonucleotides when hybridized to a complementary target only when the nucleotides are perfectly base-paired at the junction. Oligonucleotide products are exponentially amplified by thermal cycling of the ligation reaction in the presence of a second set of adjacent oligonucleotides, complementary to the first set and the target. A single-base mismatch prevents ligation and amplification, thus distinguishing a single base mutation from the normal allele. The use of a thermostable ligase allows the enzyme to survive thermal cycling in a fashion analogous to Taq polymerase in the polymerase chain reaction. The assay is compatible with nonradioactive detection and has the potential for automation. Although still in its early stages of development, LCR is expected to find many uses in the field of gastroenterology and in medicine in general. In this review we briefly describe how LCR works and discuss potential areas of application in gastroenterology. PMID- 8409325 TI - Meta-analysis on the maintenance of remission in Crohn's disease. PMID- 8409326 TI - Rehabilitation medicine makes dysphagia "easier to swallow". PMID- 8409327 TI - What is the aim of esophageal variceal sclerotherapy--prevention of rebleeding or complete obliteration of veins? PMID- 8409328 TI - The stomach in cirrhosis. The legend of Proteus retold. AB - Portal hypertensive gastropathy (PHG) and gastric antral vascular ectasia (GAVE) (watermelon stomach) are increasingly recognized as separate nosological entities detectable by careful upper gastrointestinal endoscopy and meticulous histological assessment. The have a significant phenomenological overlap; both usually present with gastric mucosal hemorrhage and have a striking association with cirrhosis. However, the distinct endoscopic and histological features, which are discussed in this paper, enable physicians to differentiate PHG from GAVE. Portal hypertension as the prerequisite of PHG necessitates surgical (portosystemic shunting) or medical (beta-blockade) portal decompressive therapy, whereas the angiodysplasia-like lesions in watermelon stomach are successfully treated by electrocoagulation or laser therapy. PMID- 8409329 TI - Endoscopy-negative dyspepsia. Hold those forceps! AB - Nonulcer dyspepsia (NUD) is an enigmatic disorder which likely has heterogeneous etiologies. Helicobacter pylori-associated gastritis has been identified in a large number of patients with NUD, raising speculation that chronic infection with this organism may cause dyspepsia in some patients. Prospective case-control and cohort studies however do not suggest that H. pylori prevalence is greater in NUD than in asymptomatic controls. Limited therapeutic trials of H. pylori eradication therapy have not shown a convincing symptomatic improvement compared with placebo treatment. Definition of the role of H. pylori in NUD awaits further well-designed, well-controlled epidemiologic and therapeutic studies. Until such information is available, routine endoscopic gastric biopsies looking for H. pylori in patients with NUD should be abandoned. PMID- 8409330 TI - Pathogenesis of sodium retention in cirrhosis. PMID- 8409331 TI - Liver transplantation in primary hepatocellular carcinoma. PMID- 8409332 TI - MDR1 (multidrug resistance) gene expression in human primary liver cancer and cirrhosis. AB - MDR1 RNA levels were analysed in patients with hepatocellular carcinoma (HCC), benign liver tumors or cirrhosis. None of the patients with HCC had received chemotherapy. MDR1 RNA levels were increased relative to a normal liver specimen in 15/26 liver cancers, 2/6 benign liver tumors and 0/8 tumor-free cirrhotic livers. MDR1 was also overexpressed in cirrhotic non-tumorous liver tissues of some patients with HCC. The results indicate that the MDR1 gene is overexpressed in liver tumors and some preneoplastic lesions and that might account for the very poor response of primary liver cancers to chemotherapy. PMID- 8409333 TI - Primary biliary cirrhosis. Histological evidence of disease recurrence after liver transplantation. AB - Histological evidence of primary biliary cirrhosis (PBC) recurring after orthotopic liver transplantation (OLT) was looked for in a 'blinded' study of 353 biopsies from 188 patients, 12-100 months post-transplant. Biopsies (172) were obtained from 83 patients transplanted for PBC and 181 biopsies from 105 patients with other liver diseases. Sixteen biopsies from 13 PBC patients (16%) had features suggestive of recurrent disease. The main diagnostic findings were: mononuclear portal inflammatory infiltration (n = 16), portal lymphoid aggregates (n = 14), portal epithelioid granulomas (n = 14) and bile duct damage (n = 15). This combination of changes was not seen in any biopsy from the non-PBC group. Additional features supporting a diagnosis of recurrent disease were ductopenia (n = 7), bile ductular proliferation (n = 7), portal fibrosis (n = 6) and copper deposition (n = 5). Thirteen biopsies from 12 patients were classified as stage I or II histologically. The other patient developed progressive damage in three serial biopsies resulting in an early micronodular cirrhosis, 5 years post transplant. These observations provide further evidence that PBC recurs after OLT. More studies are required to determine the natural history and clinical significance of the predominantly early histological changes documented so far. PMID- 8409334 TI - Risk factors for cytomegalovirus and severe bacterial infections following liver transplantation: a prospective multivariate time-dependent analysis. AB - Risk factors for cytomegalovirus and severe bacterial infections were studied prospectively by univariate, multivariate and time-dependent Cox model analysis in 79 consecutive liver transplant patients treated with selective bowel decontamination. Cytomegalovirus infection occurred in 39 patients (49%) and was symptomatic in 22 patients. Twenty-six patients (33%) developed at least one of 43 documented severe bacterial infections. In a multivariate analysis of risk factors for all cytomegalovirus infections, the cytomegalovirus seronegative recipient-cytomegalovirus seropositive donor group was the highest risk group (P < 0.001). Using the same analysis for risk factors for symptomatic cytomegalovirus infections, a prolonged prothrombin time (P < 0.005), a diagnosis of acute fulminant hepatitis as the underlying liver disease (P < 0.01) and a cytomegalovirus seronegative patient receiving a liver from a seropositive donor (P < 0.001) were significant. The treatment with OKT3 therapy (P < 0.008) and hepatic artery thrombosis (P < 0.02) were found to be significant risk factors in a time-dependent univariate analysis but were not independent risk factors when multivariate analysis was utilized. Significant risk factors for major bacterial infections (P < 0.03) using univariate analysis included a prolonged anesthesia, anhepatic and surgical times, as well as the transfusion of large amounts of fresh frozen plasma or autologous blood. In a multivariate analysis, only the transfusion of large amounts of fresh frozen plasma (P < 0.04) was a significant independent risk factor. Cytomegalovirus infection was a risk factor for the development of severe bacterial infections (P < 0.03) in a multivariate time dependent analysis. PMID- 8409335 TI - Ischemic liver cell damage and calcium accumulation in rats. AB - Regions of calcium accumulation were studied by 45Ca-autoradiography and microdensitometry during liver ischemia and reperfusion in rats. In autoradiographic studies 45Ca accumulated in all zones of the liver lobuli after 1-2 h of liver ischemia and 1, 3 and 6 h of reperfusion, but did not accumulate in non-ischemic liver lobes. Significant 45Ca accumulation occurred only with reperfusion after 1 or 2 h of liver ischemia. In the presence of reperfusion, 45Ca accumulation correlated with the length of ischemia. In histopathologic studies, liver necrosis was absent in lobuli after 1-h reperfusion. Some liver necrosis was observed in the group reperfused for 3 h and widespread liver necrosis appeared after 6-h reperfusion. Regions of 45Ca accumulation coincided with sites of microscopic liver cell damage and necrosis. These results suggested that calcium accumulation might be responsible for the liver cell damage induced by ischemia with reperfusion, that the intensity of calcium accumulation in 45Ca autoradiograms indicates the degree of ischemia damage, and that the primary event leading to hepatocellular necrosis might be calcium accumulation and subsequent liver cell death. PMID- 8409336 TI - The safety and value of extradural intracranial pressure monitors in fulminant hepatic failure. AB - Thirty-six of 68 consecutive patients with fulminant hepatic failure (FHF) progressing to grade 4 encephalopathy who had extradural ICP monitors inserted were reviewed to determine the safety and the value of ICP monitoring. Only minor complications were encountered. These included local wound bleeding at the burrhole site in four patients and a small cerebral hemorrhage in relation to the monitor in one other patient. No significant long-term sequelae were related to the operative procedure. ICP monitoring identified rises in ICP unaccompanied by clinical signs and as a consequence treatment was given to the monitored patients more often than the non-monitored group (median 6 vs. 2 treatments, P < 0.01). The duration of survival from the onset of grade 4 encephalopathy was significantly greater in the ICP monitored group (median 60 vs. 10 h, P < 0.01) although overall survival was unchanged. Monitoring also provided important prognostic information since the peak ICP was higher in non-survivors than in survivors (median 45 vs. 35 mmHg, P = 0.051). The pattern of clinical signs accompanying episodes of intracranial hypertension differed between survivors and non-survivors. Pupillary abnormalities were detected more often in non-survivors while systolic hypertension occurred more frequently amongst survivors with the peak systolic blood pressure being significantly higher. ICP monitoring proved safe and effective, provided valuable information regarding subclinical intracranial hypertension and prognosis and should be regarded as part of the routine management of intracranial hypertension complicating FHF. PMID- 8409337 TI - Chronic non-A, non-B hepatitis complicated by end-stage renal failure treated with recombinant interferon alpha. AB - Chronic non-A, non-B hepatitis occurs in 50% of Saudi patients with end-stage renal failure and requires long-term hemodialysis since it is a contraindication to renal transplantation. Thirteen patients with biochemical and histological documented chronic non-A, non-B hepatitis (11 with HCV antibodies) entered a double-blind placebo controlled cross-over study, in which Roferon A 3 MU or placebo were administered subcutaneously 3 times weekly after hemodialysis for 6 months. The mean ALT fell significantly from pretreatment levels of 74.7 (95% confidence interval (CI) 54.7, 92.5) (13 patients in the 6-month run-in period) and 66.8 (CI 47.7, 85.8) (7 patients in the run-in period + 6 patients in the placebo period) (difference NS) to 37.6 (CI 21.0, 54.2) during interferon treatment (P < 0.005). In 10/13 patients (77%) ALT levels became normal. In the 6 month follow-up period immediately after therapy, the mean ALT was 45.2 (CI 28.0, 62.0). Although this change was not significant (P = 0.49), only 7 of these 10 patients sustained biochemical remission in the 6-month follow-up period. The corresponding total Histological Activity Index improved from 8.9 (CI 7.5, 10.3), 8.9 (CI 7.2, 10.7) (difference NS) to 6.2 (CI 3.9, 8.5) (P < 0.05; P = 0.052, respectively). Intralobular inflammation and periportal inflammation showed the most significant changes. Five of 13 (39%) and 2/13 patients (15%) had complete resolution of piecemeal necrosis and intralobular inflammation, respectively. Toxic effects of interferon were mild, early and self-limiting.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409338 TI - Transjugular intrahepatic portosystemic stent-shunt (TIPS) in the treatment of Budd-Chiari syndrome. AB - Budd-Chiari syndrome is characterized by splanchnic congestion due to obstruction of the hepatic venous outflow. A variety of treatment modalities have limited applicability due to their invasive nature, complications or low effectivity. The transjugular intrahepatic portosystemic stent-shunt (TIPS) offers a new treatment by creating an intraparenchymal duct between a main branch of the portal vein and hepatic vein i.e. the intrahepatic part of the inferior vena cava. This paper describes the treatment of two patients with fulminant and subacute Budd-Chiari syndrome treated 2 days and 2 months after the onset of clinical symptoms. It demonstrates that TIPS is a feasible treatment of Budd-Chiari syndrome that restores splanchnic blood flow, reduces collateral circulation and ascites and provides sufficient time to allow for elective liver transplantation, if indicated. Further studies are required to evaluate the effect of TIPS on liver function and survival. PMID- 8409339 TI - Sodium retention and hepatic function following partial portal vein ligation in the rat. AB - Recent studies have suggested that the development of sodium retention in experimental cirrhosis in the rat occurs when hepatic function, measured by the aminopyrine breath test, decreases below a critical threshold. The present study evaluated the relationship between renal sodium handling and hepatic function and determined whether sodium retention occurs following partial portal vein ligation. Sodium balance, urine volume, creatinine clearance and the aminopyrine rate constant of elimination, on a constant sodium intake, were evaluated daily, from 1 day before surgery to 5 days after surgery, in both sham-operated (n = 6) and partially portal vein-ligated rats (n = 14). In the partially portal vein ligated group, sodium retention occurred in 9 rats between 1 and 4 days after surgery, accompanied by a 45% reduction in the aminopyrine rate constant of elimination. Spontaneous natriuresis occurred within 5 days after surgery, and was associated with an increase in the aminopyrine rate constant of elimination from 0.94 +/- 0.07 x 10(-2) min-1 on the last day of sodium retention to 1.36 +/- 0.06 x 10(-2) min-1 on the day of diuresis (P < 0.05). In contrast, creatinine clearance did not change throughout the study. There was a negative curvilinear association between sodium balance and the aminopyrine rate constant of elimination (r = 0.70, P < 0.001). In the five rats without sodium retention, there was no change in the aminopyrine rate constant of elimination and creatinine clearance over the 5 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409340 TI - Complete nucleotide sequence and viral envelope protein expression of a hepatitis B virus DNA derived from a hepatitis B surface antigen-seronegative patient. AB - An HBV strain isolated from a patient lacking conventional serological markers of HBV infection was characterized. The complete nucleotide sequence was determined following PCR amplification. Only 22 nucleotide substitutions were found relative to the reported sequence of the ayw subtype. Five of these point mutations in the preS/S and preC/C genes led to amino acid substitutions and, with one exception, were located in regions coding for antigenic determinants of viral envelope or capsid proteins. Eight amino acid substitutions were located in terminal protein and the spacer domain of the polymerase gene product. Despite these amino acid changes, transient expression of the preS2 and S envelope proteins in eucaryotic cells yielded viral proteins detectable in the culture medium with polyclonal and monoclonal anti-preS2 and -S antibodies. These data conclusively demonstrate persistent infection by HBV in subjects without HBV serological markers. The absence of conventional HBV serological markers is probably due to several factors: a low level of viral replication, some genetic modifications, as well as an abnormal immune response to the virus. PMID- 8409341 TI - Foscarnet treatment of chronic hepatitis B in an HIV-positive patient. PMID- 8409342 TI - Acute severe herpes simplex hepatitis with virus-associated hemophagocytic syndrome in an immunocompetent adult. PMID- 8409343 TI - Gilbert's disease: a risk factor for paracetamol overdosage? PMID- 8409344 TI - A case of portal vein thrombosis associated with protein S deficiency. PMID- 8409345 TI - Acute exacerbations in chronic HBsAg carriers: is HCV implicated? PMID- 8409346 TI - Hospital-acquired bacterial infections in patients with cirrhosis undergoing selective intestinal decontamination. PMID- 8409347 TI - Breast health promotion: behaviors of midlife women. AB - The breast is one of the leading sites of invasive cancer. The purpose of this study is to examine the relationships between health locus of control, perceived self-efficacy, demographic health characteristics, and the degree to which they explain the current and future practice of breast self-exam and mammography screening. Data were collected during 1992. Eighty-six women participating in a clinical breast exam and mammography screening program at an ambulatory women's center in a large metropolitan teaching hospital in the Midwest voluntarily completed four questionnaires. Hierarchical regression and discriminant analysis demonstrated that perceived self-efficacy was a significant predictor of health promotion behavior, and health-promoting lifestyle and locus of control were the most influential predictors of intent for breast self-examination and ongoing mammography. PMID- 8409348 TI - Health promotion practices of pregnant and nonpregnant women. AB - The purpose of this research was to study the health-promoting practices of pregnant and nonpregnant women and to examine the differences between the two groups. A descriptive correlational design was employed for this study. The Health Promotion Lifestyle Profile was used to measure health-promoting practices in the sample populations. Although there were no significant differences between health-promoting practices of pregnant and nonpregnant women in most areas, pregnant women scored significantly higher on the Nutrition subscale. This research suggests that all women need to increase the number of their health promoting practices. PMID- 8409349 TI - The maturing process: a vital element in the holistic care of women. AB - The maturing process is a powerful holistic method for removing childhood blocks, thus freeing us to develop our full potential in the present. This health patterning modality is an imagery process that assists us in reshaping the holograms in our memory banks. This process is effective with women coming from a variety of dysfunctional backgrounds, dealing with codependency, sexual abuse, and grief and loss issues. Female relationships and interactions are explored through the use of two case studies. The maturing process represents a missing and vital element in the holistic care of women. PMID- 8409350 TI - A holistic nursing response to mastectomy trauma syndrome. AB - As a consequence of mastectomy, women may experience disruptions in four domains of life: the physical, the psychological, the social, and the spiritual. Mastectomy trauma syndrome refers to these lifestyle disruptions. This ethnographic research study explores the major lifestyle disruptions associated with mastectomy. Holistic concepts and strategies are suggested to assist postmastectomy women toward self-actualization. PMID- 8409351 TI - An exploratory study of the effectiveness of a relaxation with guided imagery protocol. AB - An exploratory study was done to examine the effectiveness of a relaxation with guided imagery protocol in reducing anxiety and depression and increasing self esteem in new mothers during the first 4 weeks postpartum. The results indicate that anxiety and depression declined and self-esteem increased in subjects in both the experimental and control groups over the 4-week period. However, the decline in anxiety and the increase in self-esteem was greater in the experimental group than in the control group. The decline in depression was about the same in both groups. Further study, using more precise methodology and instrumentation, needs to be done to verify the results. PMID- 8409352 TI - Healing touch as a nursing intervention: wound infection following cesarean birth -an anecdotal case study. AB - From the context of postoperative wound infection care, this article describes the clinical use of Healing Touch, an integrated system of energetic healing techniques now being taught nationwide in cooperation with the American Holistic Nurses' Association. The author documents the progress of a patient with a significant wound infection as a consequence of cesarean birth. Along with standard medical care, the patient received Healing Touch treatments. This is an anecdotal report and is not intended to replace a structured study on wound healing. PMID- 8409353 TI - Menstrual-cycle lengthening and reduction in premenstrual distress through guided imagery. AB - This study investigated the effects of a program of relaxation and specific guided imagery on menstrual-cycle length and premenstrual distress. Thirty healthy college women with regular menstrual cycles were studied for 6 months. The subjects completed the Menstrual Distress Questionnaire (MDQ) at the beginning and end of the study and recorded their menstrual cycles for 3 months on an investigator-developed calendar recording sheet. Subjects were then given an audiotape with a progressive muscle relaxation exercise followed by guided imagery with a suggestive message focusing on lengthening the menstrual cycle and delaying the onset of menstrual bleeding. The 15 subjects who completed the entire study had significant increases in cycle lengths during the 3 months of imagery. The total premenstrual distress scores also declined significantly, as did the subscales measuring behavior and negative affect. This study provides preliminary evidence that menstrual-cycle rhythmicity and premenstrual distress are amenable to the mind-body intervention of guided imagery and suggests that further investigation of this phenomena with larger sample size and careful controls for confounding variables be conducted. PMID- 8409354 TI - The role of sex in bacterial evolution. PMID- 8409355 TI - The mutational meltdown in asexual populations. AB - Loss of fitness due to the accumulation of deleterious mutations appears to be inevitable in small, obligately asexual populations, as these are incapable of reconstituting highly fit genotypes by recombination or back mutation. The cumulative buildup of such mutations is expected to lead to an eventual reduction in population size, and this facilitates the chance accumulation of future mutations. This synergistic interaction between population size reduction and mutation accumulation leads to an extinction process known as the mutational meltdown, and provides a powerful explanation for the rarity of obligate asexuality. We give an overview of the theory of the mutational meltdown, showing how the process depends on the demographic properties of a population, the properties of mutations, and the relationship between fitness and number of mutations incurred. PMID- 8409356 TI - The evolution of sex and recombination in a varying environment. AB - Theoretical models of the evolutionary advantages of sex and genetic recombination in a temporally varying environment are analyzed. The models assume a quantitative trait controlled by many additive genes and subject to nor-optimal selection. The optimal value of the trait is assumed either to move steadily in one direction or to follow an autocorrelated linear Markov process. The consequences for population mean fitness of a reduction in genetic variance, due to a shift from sexual to asexual reproduction, are examined. It is shown that a large reduction in mean fitness can result from such a shift in the case of a steadily moving optimum, under light conditions. The conditions are much more stringent with a randomly varying environment, especially if the autocorrelation is low. The conditions for spread of a rare modifier affecting the rate of genetic recombination are also examined, and the strength of selection on such a modifier is determined. Again, the case of a steadily moving optimum is more favorable than a random optimum for the evolution of increased recombination. PMID- 8409357 TI - The sexual nature of the eukaryote genome. AB - This paper supports a previous conjecture that the sexual cycle of eukaryotes arose from the infection of cells by genome parasites. The finding are as follows. (1) In prokaryotes, conjugative plasmids ensure their own spread by directing partial cell fusion. (2) Conjugative plasmids permit gene transfer between prokaryotes and eukaryotes, and can be integrated into the eukaryote genome. (3) Genes can be transferred between unrelated eukaryotes, and between different genomes within eukaryotic cells. (4) Elements such as transposons and retroviruses evolve as parasites of the eukaryote sexual system. (5) The mating type genes of bipolar fungi are idiomorphic: alternative genes directing sexual specificity are dissimilar and non-homologous. It is argued that they arose as parasitic elements directing cell fusion, which have become integrated into the genome. (6) Mating-type idiomorphs in different taxa may be dissimilar, reflecting the independent acquisition of different infectious elements by different eukaryote lineages. (7) Sexual competence is rapidly lost through mutation accumulation or antagonistic pleiotropy during vegetative proliferation, so that many lineages are sexually sterile. (8) In multipolar systems, novel mating-type alleles arising by mutation spread in a parasite-like manner by virtue of their access to lineages which have become sexually sterile. (9) It is suggested that centromeres arise as devices to enable low copy number plasmids to persist. (10) Crossing over is favored if it enables the genes that direct it to unlink themselves from inferior genomes, or from the consequences of their own transposition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409358 TI - Genetic error, sex, and diploidy. AB - Mathematical models and experiments on transformation are reported testing the hypothesis that sex and diploidy evolved as a DNA repair system. The models focus on the origin of diploidy and sex by studying selection between asexual haploids, sexual haploids, and diploids. Haploid cells are efficient replicators, while diploid cells are resistance to damage. A sexual haploid may combine the advantages of both: spending much of its life cycle in the haploid state, then temporarily fusing to become diploid, followed by splitting to the haploid state. During the diploid state DNA damage can be repaired, since there are two copies of the gene in the cell and one copy is presumed to be undamaged. Five basic rate parameters are employed: birth and death; genomic damage (for the haploids alone); and, for the sexual cell, fusion and splitting. Parameter space bifurcation diagrams for the equilibria are drawn, and solutions of the equations are described in terms of these diagrams. Each type of cell has a region of the parameter space that it occupies exclusively (given its initial presence in the competition). The haploid wins in environments characterized by low damage. The diploid wins in environments characterized by high damage, low mortality, and abundant resources. In general, only a single type of cell occupies a given portion of the space. We find, however, that competitive coexistence of an asexual diploid and sexual haploid is possible in spite of the fact that they are competing for a single resource (nucleotide building blocks). Sex can increase from rarity if matings occur with asexual cells. Only sex can cope with both high mortality and high damage. We then turn to natural bacterial transformation as a model system for the experimental study of sex. Natural transformation in distributed widely, but apparently sparsely, in all bacterial groups. A very preliminary phylogenetic analysis of the bacilli and related species indicates that transformation is probably not a diversifying force in bacterial evolution. However, it is difficult to be sure because of the ambiguity surrounding negative data. Experiments with the bacterium Bacillus subtilis indicate that transformation frequencies respond adaptively to DNA damage if homologous donor DNA is used. Several specific hypotheses for this response are considered. Recent work in other labs on the evolution of transformation is discussed from the point of view of the hypothesis that transformation functions in DNA repair. PMID- 8409359 TI - Classification of hypotheses on the advantage of amphimixis. AB - A classification of hypotheses on the advantage of amphimixis over apomixis is presented. According to "Immediate Benefit" hypotheses, amphimixis is advantageous regardless of reciprocal gene exchange, because either it directly increases fitness of the progeny, reduces the deleterious mutation rate, or makes selection more efficient. In contrast, "Variation and Selection" hypotheses attribute the advantage of amphimixis to the reciprocal gene exchange that alters genetic variability and response to selection among the progeny. Most such hypotheses assume that amphimixis increases variability and efficiency of selection, but some claim that amphimixis decreases response to selection. Variation and Selection hypotheses require that some factor, either random drift or epistatic selection, makes distributions of different alleles nonindependent, while another factor, either changes of the genotype fitnesses or deleterious mutations, makes overrepresented genotypes non-optimal. Numerous Variation and Selection hypotheses, dealing with either unstructured or spatially structured populations, are reviewed. Two of them seem most plausible: better responsiveness of the amphimictic population to widely fluctuating selection, and lower mutation load in the amphimictic population under synergistic selection against deleterious mutations. In both cases the large advantage of amphimixis requires rather stringent conditions, which could be falsified by careful experiment. Further progress in understanding the evolution of amphimixis will depend mostly on such experimental work. PMID- 8409360 TI - Genes for breakfast: the have-your-cake-and-eat-it-too of bacterial transformation. AB - Bacterial transformation, in which cells take up and recombine free strands of DNA, is the simplest process thought to have evolved for genetic exchange, i.e., because of the potential benefits of producing progeny with recombinant genotypes. However, two other functions are equally plausible: acquisition of intact DNA strands to use for recombinational repair of DNA damage, and acquisition of the nutrients contained in DNA molecules. Although the recombinant progeny produced by transformation can be beneficial, the success of genes causing transformation is limited by other factors, especially by the genetic quality of DNA derived from dead cells. Our recent experiments in the naturally transformable bacteria Haemophilus influenzae and Bacillus subtilis suggest that the DNA-repair hypothesis is unlikely to be correct. In H. influenzae, transformation does not detectably increase the cells' ability to survive DNA damage. More importantly, we have found that, although competence (the ability to take up DNA) is induced by nutritional limitation in both H. influenzae and B. subtilis, it is not induced by DNA damage in either. Thus we favor the hypothesis that transformation evolved as a nutrient-uptake system, especially because unrelated DNA is abundant in the environments of many naturally transformable bacteria. PMID- 8409361 TI - Chromosomal and mitochondrial DNA variation in orang utans. AB - Wild-born orang utans held in zoos do not have their geographic origins defined, which has complicated comparisons of the two subspecies of orang utans (Bornean orang utans, Pongo pygmaeus pygmaeus, and Sumatran orang utans, P. pygmaeus abelii). The resulting confusion concerning the subspecies identity of individual orang utans and a larger controversy over the desirability of utilizing subspecies designations in orang utan breeding programs has focused attention on the need for a phylogenetic analysis of the species and an examination of the appropriateness of the subspecific designations. Previous studies have suggested that an inversion in the second chromosome pair (PPY2) is a diagnostic subspecies marker and that mitochondrial DNA (mtDNA) and nuclear genetic markers may also be used to identify the subspecies origin of individual orang utans. In an effort to assist zoological parks in examining the question of genetic divergence of orang utan subspecies, we have studied the chromosomes of 144 orang utans, including 58 wild-born individuals. To gain insight into phylogenetic divisions of these apes, mtDNA restriction cleavage site variation has been investigated in 14 individuals whose karyotypic status was known. These investigations have confirmed the existence of two phylogenetic lineages of orang utans based on mtDNA cleavage patterns and demonstrated that these lineages correspond with the two populations characterized by the G-banded morphology of the second pair of chromosomes. The two orang utan phylogenetic units comprise individuals that generally conform to the recognized allopatric, morphological subspecies. Based on current knowledge, the view that the named orangutan subspecies each constitute a distinct phylogenetic lineage more consistent with species-level divergence is supported. PMID- 8409362 TI - Molecular symbionts and the evolution of sex. AB - The main focus of this article is on the evolutionary origin of sex rather than its maintenance in extant organisms. Sexual outbreeding involves a complex set of biological phenomena, and it is useful to consider the probable order in which the components of this process evolved. I propose that conjugation between cells was the initial stage in the evolution of sex. It has been shown that molecular symbionts (such as transposons and plasmids) derive a major selective advantage from conjugation and sexual outbreeding. This strongly suggests that the origin of conjugation between cells may best be understood as a symbiont-encoded adaptive function. In other words, conjugation between haploid cells evolved initially as a means of propagating molecular symbionts. Once efficient mechanisms of conjugation were in place, however, the subsequent elaboration of the other aspect of sex, particularly genetic recombination, were facilitated. Although the molecular symbiont theory is directly relevant only to the origin of sex, it also has implications for understanding the subsequent stages in the evolution of sex. PMID- 8409363 TI - The effects of different levels of genetic exchange on organelle evolution. AB - Each mitochondrion and chloroplast organelle has tens to hundreds of genomes, and there are tens to hundreds of such organelles within each cell. This results in a genetic organization generating a population structure akin to a geographically structured population. Since genomes and organelles replicate and gene conversion can occur between copies, this offers the possibility for directional evolutionary forces acting within the cell. The consequences of genetic exchange within the cell (between genomes within an organelle and between individual organelles) and between individuals (biparental inheritance) when within-cell directional forces are present is examined. The net effect of genetic exchange is to increase the importance of any directional forces present within the cell. The consequences of within-cell forces when the variants they act on also have fitness consequences for individuals are also examined. PMID- 8409364 TI - Rewriting medical history: Charles Best and the Banting and Best myth. PMID- 8409365 TI - Cholera and plague in India: the bacteriophage inquiry of 1927-1936. PMID- 8409366 TI - The enigma of facial expression: medical interest in metoposcopy. PMID- 8409367 TI - The function of praise in the contract of a medieval public physician. PMID- 8409368 TI - Ultrastructural immunogold localization of tumor necrosis factor-alpha to the matrix compartment of eosinophil secondary granules in patients with idiopathic hypereosinophilic syndrome. AB - Peripheral blood eosinophils from two normal donors and two patients with the hypereosinophilic syndrome (HES) were analyzed with a post-embedding immunogold method to detect the substructural location of tumor necrosis factor-alpha (TNF alpha). In eosinophils of HES patients, TNF-alpha was localized to the matrix compartment of 64% of the specific secondary granules. Other structures in the HES eosinophils were unlabeled. No TNF-alpha was detected in eosinophils of normal donors. These studies document the first ultrastructural subcellular localization of any cytokine within the major population of secretory granules in human eosinophils and support other lines of evidence indicating that the expression of TNF-alpha may be greater in the eosinophils of HES patients than in those of normal donors. PMID- 8409369 TI - Localization of amidating enzymes (PAM) in rat gastrointestinal tract. AB - We studied the distribution of the two enzymes involved in post-translational C terminal alpha-amidation of regulatory peptides in rat digestive tract, using immunocytochemical methods and in situ hybridization techniques. The enzymes were located in most of the fibers and neurons of the myenteric and submucous plexus throughout the entire digestive tract and in endocrine cells of the stomach and colon. Staining of reverse-face serial sections demonstrated that the enzymes in endocrine cells of the stomach co-localized with gastrin in the bottom of the gastric glands. Some gastrin-immunoreactive cells near the neck of the gland were negative for PAM, suggesting that amidation takes place only in the more mature cells. In the colon all cells immunoreactive for glucagon and GLP1 were also positive for peptidylglycine alpha-hydroxylating monooxygenase (PHM) but not for peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL). The absence of immunoreactivity for the amidating enzymes in endocrine cells of the small intestine, known to produce C-terminally amidated peptides, suggests the existence of other amidating enzymes. PMID- 8409370 TI - Monoclonal antibodies to the Golgi apparatus of serous exocrine cells. AB - We demonstrated that a common antigen (Golgi-associated antigen, GAA 108) is present in the Golgi apparatus of serous exocrine cells, using an immunohistochemical method with monoclonal antibodies (MAb) 108 (IgG1) and 18 (IgM), raised to the microsomal fractions of rat parotid gland. The MAb reacted with polypeptides of molecular weights in the 58-170 KD range in parotid gland on Western blot analysis. The Golgi apparatus of the following cells was immunostained with these MAb: acinar cells of parotid gland, pancreas, and exorbital lacrimal gland, serous cells of sublingual gland, chief cells of stomach, and epithelial cells of rat prostate. However, positive reaction occurred throughout the entire cytoplasm of submandibular gland acinar cells. Immunoelectron microscopy (IM) revealed antigen (GAA 108) localization in the medial and trans-Golgi cisternae and trans-Golgi network (TGN), including condensing vacuoles, in parotid, exorbital lacrimal, and pancreatic acinar cells, and serous acinar cells of sublingual gland. Lysosomes and apical cell membranes also stained positively in some cells. In the submandibular gland reactions were observed in the medial and trans-Golgi cisternae, condensing vacuoles, secretory granule contents, cell membrane, and in some duct lumens. These results suggest that although GAA 108 is found in the Golgi apparatus of most serous exocrine cells, it is secreted by a regulated pathway in the acinar cells of submandibular gland. PMID- 8409371 TI - Combined immunocytochemistry and enzyme cytochemistry on ultra-thin cryosections: a new method. AB - We combined immunocytochemistry and enzyme cytochemistry to localize two different proteins on the same ultrathin cryosection. In this method the immunocytochemical localization is visualized with colloidal gold probes and the enzyme cytochemical detection is achieved with cerium as the capture agent. The immunocytochemistry is conducted first so that any potential adverse effects of the enzyme cytochemical procedure will not alter the antibody binding properties of the cryosections. PMID- 8409372 TI - Localization of mRNAs of two androgen-dependent proteins, SMR1 and SMR2, by in situ hybridization reveals sexual differences in acinar cells of rat submandibular gland. AB - Androgen-dependent sexual differences in the granular convoluted tubules of mouse and rat submandibular glands (SMG) have been extensively reported. We studied two major androgen-dependent mRNAs of the rat SMG encoding proteins named SMR1 and SMR2. To determine which cell type in the SMG is responsible for synthesis of these mRNAs, we performed in situ hybridization with digoxigenin-labeled RNA probes coupled with alkaline phosphatase detection. We show that SMR1 and SMR2 mRNAs are synthesized in the acinar cells of the SMG. A clear difference in SMR1 and SMR2 mRNA levels in male and female is demonstrated. During the course of this study we also confirmed the acinar localization of mRNAs encoding the glutamine/glutamic acid-rich proteins (GRP) of rat SMG. Our data are the first clear evidence of androgen-dependent sexual differences in acinar cells of rat submandibular gland. PMID- 8409373 TI - Selective accumulation of diamidino yellow and chromomycin A3 by retinal glial cells. AB - We applied the fluorescent DNA stains diamidino yellow (DY) and chromomycin A3 to rat and rabbit retinas in vivo and in vitro. They accumulated in the nuclei of a subpopulation of cells of the inner nuclear layer. The number and distribution of the fluorochrome-accumulating cells were similar to those of the Muller glia, and double-labeling experiments showed that the cells accumulating DY or chromomycin A3 contained oriented filaments of vimentin. The fluorochromes also accumulated in the sparse astrocytes and oligodendrocytes located among the myelinated fibers of the rabbit central retina. Specific accumulation in retinal glia occurred only when the fluorochromes were applied to living retinas. If the plasma membranes were disrupted by fixation or exposure to detergent, most retinal cells were stained. This indicates that the locus of specificity is the entry of the molecules into the cells. When applied to living retinas, other DNA stains selectively accumulate in subclasses of retinal neurons. Why DNA-binding molecules should selectively cross the membranes of either retinal neurons or retinal glia remains an unsolved problem. PMID- 8409374 TI - Ultrastructural and antigenic preservation of a delicate structure by cryopreparation: identification and immunogold localization during biogenesis of a secretory component (membrane-matrix connection) in Paramecium trichocysts. AB - Ultrastructure and antigenicity of the "mesh-like sheath" (MLS), a very delicate structure connecting the membrane and the paracrystalline matrix of Paramecium trichocysts, are well preserved after cryofixation (rapid freezing followed by freeze-substitution in methanol and embedding in Lowicryl K11M at 213K). The MLS is labeled by colloidal gold-bound antibodies (Ab-Au10nm) with primary antibody (Ab) against trichocyst components obtained by recloning hybridoma cells twice. We prepared Western blots from reduced gels obtained from subfractionated trichocysts. Trichocyst membranes displayed reactive bands of 68-70, 63-66, 43, 40 (strongest), and 57 and 54 KD, with a weak band of 38 KD. One of the most abundant protein bands of soluble secretory components (56-57 KD) was also strongly stained on blots. On ultra-thin sections pre-trichocysts display Ab reactive material concentrated below the trichocyst membrane before the MLS can be recognized as a structural entity. Quantitative evaluation of Ab-Au10-labeled ultrathin sections also revealed passage of MLS materials through the very inconspicuous Golgi apparatus. This was substantiated by Ab-peroxidase labeling. We conclude that MLS components (whose ultrastructure is difficult to preserve) are largely membrane-associated, partly soluble proteins. They form a connection (released during exocytosis) between the abundant paracrystalline matrix components and the organelle membrane. MLS might thus maintain a peripheral aqueous space of functional importance. PMID- 8409375 TI - Immunohistochemical localization of proteoglycans in human periodontium. AB - Proteoglycans (PGs) are extracellular and cell surface-associated macromolecules that regulate cell adhesion, cell growth, matrix formation, and bind growth factors. In this work we studied the distribution of core proteins of four PGs (decorin, biglycan, a large molecular weight PG, and CD44) in human gingiva and periodontal ligament by immunohistochemical staining of frozen tissue sections with specific antibodies. Decorin, a major PG of this tissue, was localized on collagen fiber bundles in the gingival and periodontal connective tissues. Staining for decorin was most intense at the subepithelial region. Biglycan was a minor PG component of the human periodontium, showing some accumulation in connective tissue under the oral epithelium. At the immunohistochemical level, biglycan appeared to form fine filament-like structures on extracellular matrix fibers. Localization of large molecular weight PG differed from that of decorin and biglycan. It was concentrated in deep connective tissue areas of the gingiva and in the periodontal ligament, and was only weakly present at the subepithelial region. CD44 was mainly concentrated in cell-cell contact areas of basal and spinous layers of oral epithelium. In the connective tissue of gingiva and periodontal ligament, CD44 was localized on fibroblast cell surfaces. Connective tissue area under the junctional epithelium contained relatively small amounts of PGs. The results indicate that different parts of human periodontium contain a typical variety of PGs, suggesting a specific function for each PG species in the location at which they accumulate. PMID- 8409376 TI - Monoclonal antibodies to type VI collagen demonstrate new tissue augmentation of a collagen-based biomaterial implant. AB - We developed a panel of highly specific monoclonal antibodies (MAb) to either human or dog collagen Type VI. Various degrees of species crossreactivities were found with ELISA and immunohistology. Because of these differentiating species specificities, which allowed distinction between the original donor collagen and newly formed host collagen, the MAb proved to be valuable tools in examination of explanted samples of an ovine composite vascular prosthesis retrieved from a canine model. With an MAb that reacts with dog but not sheep collagen Type VI, newly synthesized pockets of collagen Type VI could readily be detected within the prosthesis as early as 3 months after implantation. These areas were associated with regions of cell infiltration, presumably derived from the host. This association was also apparent in the newly formed intimal region of the prosthesis where only host cells were found. Another of the MAb, which was positive against human but not sheep collagen, was also used to demonstrate marked deposition of host collagen Type VI in a retrieved human sample of the prosthesis. In this case the antibody was able to detect collagen on a formalin fixed tissue, which would broaden the scope of its use in clinical and pathological situations. Use of these novel antibody probes provides a rapid marker for new tissue augmentation of implanted biological devices which would be an indicator of the long-term performance of a prosthesis. PMID- 8409377 TI - Distribution of alpha 6 integrin subunit in developing mouse submandibular gland. AB - We examined the development of mouse submandibular gland by light and electron microscopy and determined the distribution of the alpha 6 integrin subunit and laminin in this process by immunofluorescence and immunoelectron microscopy. At Days 13.5 and 14 of gestation alpha 6 was localized over the entire cell surface of undifferentiated epithelial cells of the terminal cluster. On Day 15 the expression of alpha 6 could no longer be detected over central cells in the proximal portion of branched terminal cluster, whereas peripheral cells were stained over the entire surface. On Day 17 of gestation to day of birth, alpha 6 expression was restricted to the basolateral surface of the differentiated acinotubular structure. Its expression on acinar cells was uniformly distributed throughout the basolateral membrane, but on duct cells stronger staining towards the basal surface was noted. Similar expression was observed in adult acinar and duct cells. Expression of the alpha 6-subunit at the cell-cell contact in an early stage and its expression at the basolateral surface in an advanced stage indicate that integrin containing alpha 6 plays a significant role in cell-cell and cell-substrate interaction. Stage and cell type-specific change of integrin expression may be significant for submandibular development. PMID- 8409378 TI - Peptide-induced changes in class I heavy chains alter allorecognition. AB - Class I molecules of the MHC are intimately involved in the development and function of CD8+ T cells. Small peptides, derived from endogenous proteins, bind within the Ag binding groove created by the beta-pleated sheets and alpha-helices of the alpha 1 and alpha 2 domains of the class I molecule. This peptide-MHC complex has been shown to influence allorecognition by CD8+ T cells. However, the precise role of peptide in alloantigen recognition remains unclear. We have previously shown that conformational changes induced in the class I molecules can be identified as specific alterations in serologic epitopes. These results suggested that alloreactive T cells may detect structural changes in MHC based on the nature of the peptide binding to the class I protein. Here, we have shown that, in at least some instances, alloreactivity may not depend on the recognition of a precise self-peptide but on an epitope on the class I molecule influenced by the peptide. The nature of specific peptides expressed by class I bearing cells may, therefore, have a dramatic effect on T cell development, self tolerance, and alloreactivity. PMID- 8409379 TI - Clonal deletion of V beta 5+ T cells by transgenic I-E restricted to thymic medullary epithelium. AB - A variety of cell types expressing MHC class II molecules is known to function as APC in vitro. We employed the Ig kappa gene enhancer and promoter to target the class II E alpha gene, and thereby I-E, exclusively to B cells to address their APC function in vivo. Although transgenic I-E was expressed on B lymphocytes, we unexpectedly obtained I-E on thymic medullary epithelium but not macrophages and at low frequency on dendritic cells. Using these transgenic mice, we constructed bone marrow irradiation chimeras with I-E expressed only on medullary epithelium, in order to determine the role of this cell type in tolerance by clonal deletion in the thymus. Although it is accepted that bm-derived cells play a primary role in deletion, and thymic epithelium can delete clones to a lesser degree, the role of cortical vs medullary thymic epithelium has not been directly dissected. We demonstrate that medullary epithelium alone can tolerize by partial deletion of I E-reactive V beta 5+ T cells. These results indicate a role for medullary epithelium in deletion during the later stages of thymic development, and support the notion that positive and negative selection of developing T cells can occur in distinct temporal and anatomic compartments. PMID- 8409380 TI - A conservative mutation in a class I MHC molecule outside the peptide binding groove stimulates responses to self peptides. AB - A transgenic mouse has been made that expresses a mutant MHC class I H-2Kb molecule with glutamic acid at position 65 (E65) in place of glutamine. The side chain at position 65, on the outward face of the alpha-helix of the alpha 1 domain of the class I molecule, interacts with the TCR, and not with the peptide binding groove. The transgenic mouse, on a DBA/2 background, mounts Kb,E65 restricted Ag-specific responses to conventional Kb-restricted Ag such as OVA and vesicular stomatitis virus, and shows strong alloreactivity to wild-type Kb. The transgenic mouse also mounts a primary in vitro alloreactive response directed to a mutant molecule with aspartic acid at position 65 (D65). This response is relatively weak, probably because of the structural similarities between aspartic and glutamic acid side chains; both have carboxylic termini, and the aspartic acid side chain is shorter by a single secondary carbon. The alloreactive CTL lines elicited by this conservative change are cross-reactive among several position-65 variants of H-2Kb. Individual CTL clones are specific for self peptides that can be extracted from cells expressing Kb,E65, and from purified wild-type Kb molecules, and that are recognized in the context of the D65 residue. Thus, the smallest variance from self in a class I molecule, even outside the peptide binding groove, can be antigenic. PMID- 8409381 TI - Immunogenic peptides bind to class II MHC molecules in an early lysosomal compartment. AB - Exogenous protein Ag are processed within endocytic compartments to produce peptides that bind to class II MHC (MHC-II) molecules for presentation to T cells. We have now identified a subcellular compartment in which immunogenic peptides bind to MHC-II as a subset of high density lysosomes. Immunoelectron microscopy of whole cells and dense Percoll gradient subcellular fractions showed early tubulovesicular lysosomes with high levels of MHC-II. Typical mature lysosomes contained less MHC-II. Pulse-chase biosynthetic labeling of macrophages followed by immunoprecipitation of MHC-II from dense lysosomal fractions showed that MHC-II molecules targeted efficiently to lysosomes after biosynthesis. Moreover, lysosomal MHC-II molecules were rapidly loaded with immunogenic peptide (as detected by T cells) soon after exposure of macrophages to Ag and before similar expression of peptide-MHC-II complexes on the plasma membrane; this loading was blocked at 18 degrees C. We propose that nascent MHC-II molecules target to early tubulovesicular lysosomes and bind immunogenic peptides therein; the resulting peptide-MHC-II complexes are then transported to the plasma membrane. PMID- 8409382 TI - Immune functions of tumor necrosis factor. I. Tumor necrosis factor induces apoptosis of mouse thymocytes and can also stimulate or inhibit IL-6-induced proliferation depending on the concentration of mitogenic costimulation. AB - Murine rTNF produces at least three effects on mouse thymocytes in vitro: 1) Is a modest co-stimulator of proliferation with low PHA-P doses. 2) Has a bi directional interaction with rIL-6-depending on PHA concentration: at low PHA (5 to 10 micrograms/ml) TNF augments and at high PHA (20 to 30 micrograms/ml) inhibits IL-6-induced proliferation. A comparable bidirectional PHA dose dependent TNF interaction was seen with IL-1 beta, whereas only inhibition at high PHA with IL-2 and only augmentation at low PHA with IL-4 were seen. 3) TNF induces direct thymocyte apoptosis (a property not shared by IL-1 beta, IL-2, IL 4, IL-6 and IL-7). Of the cytokines studied, only IL-7 reduced TNF apoptosis. Thymocyte apoptosis by TNF showed the same species specificity as costimulation (i.e., human TNF had no effect) and was not inhibited by CY. The thymocyte CD4 CD8 phenotype after 72-h cultures showed that TNF decreased mainly double negative (DN) and single positive (SP) subsets, whereas IL-6 with low or high PHA increased DN and SP, especially the SP CD8+ subset. The regulatory and apoptotic effects of TNF were seen only with thymocytes and not with peripheral splenic or lymph node T cells. Four mAb to mouse TNF (2E2, XT22, 1C6, and 1OD9) could abrogate TNF costimulation and the TNF effects on IL-6-induced thymocyte proliferation, at both augmenting and inhibitory PHA conditions. However, only the two antibodies that also neutralize TNF lytic activity (2E2, XT22) could inhibit TNF-mediated apoptosis, implying two different but neighboring functional domains in the TNF molecule mediating apoptosis/lysis and costimulation. Our studies show that TNF might have unique and complex regulatory effects on growth and death of thymocyte populations in adult mice quite different from its effects on T cells in periphery. PMID- 8409383 TI - Multiple pathways are involved in the extracellular processing of MHC class I restricted peptides. AB - T cell stimulation by certain class I-restricted antigenic peptides, such as the HIV 1 gp160-derived peptide, P18, requires peptide processing by angiotensin-1 converting enzyme (ACE) in FCS. We observed that longer versions of P18 and the murine cytomegalovirus pp89-derived core peptide, pMCMV, which could stimulate T cell hybridomas in FCS, were not as sensitive to the ACE inhibitor captopril as P18. Using cell-free soluble murine class I MHC molecules and protease inhibitors, we found that there are pathways of differing efficiency that use enzymes other than ACE for the proteolytic processing of peptides in serum. The kinetics of the generation of T cell stimulatory activity among P18 variant peptides in serum differed with peptide length, and with the nature of amino and COOH-terminal extensions. Such processing occurs in human plasma as well as in FCS. The understanding of this processing, its kinetics, and its inhibitors can lead to better design of peptide-based therapies, including vaccines. PMID- 8409384 TI - Characterization of murine T cell responses to peptides of the variable region of self T cell receptor beta-chains. AB - Immunization with peptides of the TCR may regulate cellular immune responses that are dominated by a particular TCR. However, no extensive study of the immunogenicity of peptides of different regions of the V beta chain of the TCR has been done. We have tested the immunogenicity of several V beta peptides in several strains of mice and characterized the cellular response to these peptides. We examined the ability of six strains of mice of H-2b, H-2d, or H-2k and of mouse lymphocyte stimulatory (Mls)-1a or Mls-1b haplotypes to respond to several peptides of the V beta 6 region and an NH2-terminal peptide of other V beta of the TCR. These include V beta 6 peptides 1-20, 32-48, 39-60, 48-75, 58 75, and V beta 3, V beta 8.1, and V beta 8.3 peptides 1-20. The various mouse strains respond to these peptides independently of deletion of V beta 6+ T cells from peripheral lymphocytes. All of the Mls deleting and nondeleting strains tested respond weakly to one peptide of V beta 6, V beta 6(39-60). Antibody titers were also demonstrated in BALB/c and DBA/2J to V beta 6(1-20), V beta 6(39 60) and V beta 6(48-75), but not to V beta 6(32-48). We demonstrated that T cells responding to V beta 6(32-48) produce IL-2 and IFN-gamma, consistent with the Th1 subset of T cells. None of the antipeptide antibodies recognized the intact V beta 6 TCR on the cell surface. In vitro antibody blocking studies with TCR peptides show that these peptides are presented by class II MHC to CD4+ T cells. We conclude that the T cell and B cell repertoires contain cells able to respond to peptides of self TCR and immunization with peptides induces CD4+ T cells and that these cells may have an immunoregulatory role. PMID- 8409385 TI - Cytolytic activity of murine IL-2-producing CD4+ and CD8+ T cell clones cycles in response to IL-2. AB - Alloreactive or OVA-reactive cloned murine CD4+ or CD8+ T cells that produce IL-2 exhibit greatly reduced cytolytic activity after being cultured with high concentrations of rIL-2. Furthermore, such cells fail to produce lymphokines or proliferate when stimulated with Ag. The duration of this unresponsiveness to Ag correlates with the concentration of rIL-2 to which the cells were exposed; higher concentrations of rIL-2 prolong the period of unresponsiveness. The presence of ionomycin during the cytolytic assay restores lytic activity to cells rendered unresponsive by exposure to rIL-2. These results suggest that rIL-2 induced unresponsiveness to Ag is a consequence of impairment of a calcium dependent signal important for cytolysis, proliferation, and lymphokine production. Thus, IL-2 appears to be an important lymphokine that regulates T cell responses downward as well as upward. PMID- 8409386 TI - Heterogeneous populations of class II MHC+ cells in human dermal cell suspensions. Identification of a small subset responsible for potent dermal antigen-presenting cell activity with features analogous to Langerhans cells. AB - Little is known regarding the identification, classification, and function of class II MHC+ dendritic cells in the perivasculature of human connective tissues, such as the dermis. We developed a method for preparing papillary dermal cell suspensions from human keratome strips. Among the class II MHC+ populations of the dermis identified using triple color flow cytometry, cells of monocyte/macrophage lineage (CD45+ CD1- CD11b+ CD11clo-mid CD32+ CD36+ or - CD11a ) and mesenchymal cells of non-bone marrow origin (CD45-) were identified and characterized. Another distinct class II MHC+ subset was identified, which expressed a number of features analogous to epidermal Langerhans cells (LC) and other dendritic APC. These were a numerically minor population comprising only 2.7% +/- 1% (n = 7) of dermal cells. Like LC, they express HLA-DR, CD45, CD1a (albeit at a lower level of expression), CD1c, and CD32 and lack constitutive CD11a or ICAM-1. In contrast to LC, this dermal CD1a+CD1c+ subset expresses CD1b, CD11b, a higher level of CD11c, and intracytoplasmic factor XIIIa. Alloantigen presentation by unfractionated dermal cells was reduced by prior removal of this CD1b+ subset to the same degree achieved by removal of the entire DR+ population (20% of dermal cells), indicating that this was the critical DR+ subset. Cocultures of CD4+ T lymphocytes with cells sorted by flow cytometry into CD1c+DR+, CD1c-DR+ and DR- dermal cell subsets positively identified the CD1c+DR+ population as the most potent of potential APC subsets in human dermis. Thus, in distinction to other dermal macrophage and mesenchymal subsets with elongate morphology, the CD1aloCD1b,c+CD11c(hi)CD11b+CD32+DR+ population in human dermis is highly analogous to cells of LC/dendritic APC lineage in its phenotype and in its exclusive ability to potently present Ag to T lymphocytes. These studies identify and characterize the APC subset most potent in inducing activation of T cells initially entering the perivasculature of human dermis to be of LC/dendritic APC, and not tissue macrophage, lineage. PMID- 8409387 TI - Novel mechanism for inhibition of human T cells by glucocorticoids. Glucocorticoids inhibit signal transduction through IL-2 receptor. AB - Interaction of IL-2 with its high affinity membrane receptor complex (IL-2R) present on activated T lymphocytes induces cell proliferation and mediates effector functions. Glucocorticoids inhibit IL-2 production by inhibiting TCR mediated signal transduction. We asked whether they also inhibit the action of IL 2 by inhibiting signal transduction through IL-2R. Human peripheral blood T cells, stimulated with PMA for 48 h (PMA blasts), were incubated with IL-2 in the presence of incremental dosages of dexamethasone (Dex; 10(-5)-10(-9) M). Dex inhibited the IL-2-dependent proliferation of PMA blasts in a dose-dependent fashion (IC50, 5 x 10(-8) M). Cell surface expression of IL-2R alpha- and beta chains as determined by immunofluorescence analysis was not affected by Dex. In addition, Scatchard plot analysis of 125I-labeled IL-2 showed that Dex did not affect the binding of IL-2, thus suggesting that inhibition is due to a postreceptor effect. Inhibition of T cell proliferation by Dex was associated with decreased IL-2-dependent tyrosine phosphorylation of several intracellular proteins and decreased phosphorylation of the retinoblastoma gene product Rb, a protein essential for controlling the progression of cells through the cell cycle. IL-2-dependent IL-2R alpha expression in PMA blasts and NF-kB induction in resting human T cells were also inhibited by Dex. These results demonstrate that glucocorticoids inhibit preactivated T cells by down-regulating signal transduction through IL-2R. PMID- 8409388 TI - The TAPA-1 molecule is associated on the surface of B cells with HLA-DR molecules. AB - TAPA-1 is a transmembrane protein that has been shown to be involved in cell growth and cellular adhesion. Our studies were aimed at determining the mechanisms of the biologic phenomena mediated by TAPA-1, which include the identification of proteins that are associated with it on the surface of lymphocytes. We and others have previously shown that Leu-13, a leukocyte Ag, is one such molecule and that in B cells TAPA-1 is associated with the CD19 Ag. Herein we identify an additional molecule, HLA-DR, that is noncovalently associated on the surface of B cells with TAPA-1. This association was first detected by immunoprecipitation by anti-TAPA-1 and by anti-HLA-DR antibodies in the presence of mild detergents. The initial observation was confirmed by 2 dimensional SDS-PAGE and by direct identification of TAPA-1 in anti-HLA-DR immunoprecipitates by Western blot analysis. The association of the two molecules on the surface of a human B cell line was shown by cocapping experiments. In addition, antibodies to both molecules can induce cellular adhesion and an antiproliferative effect. Because the tissue distribution of these two molecules only partially overlaps, with TAPA-1 being expressed on most cell types and MHC class II expressed on a more restricted group of tissue, it is possible that the TAPA-1 molecule provides a basic function that can augment a cell type specific activity. In B cells the association of TAPA-1 with CD19 and HLA-DR may increase cellular interaction and play a supporting role in the transmission of specific signals. PMID- 8409389 TI - Rapid activation of C-Raf-1 after stimulation of the T-cell receptor or the muscarinic receptor type 1 in resting T cells. AB - The c-Raf-1 serine/threonine kinase is an important component of signal transduction pathways mediating the effects of a variety of growth factors. In activated T cells, IL-2 has been shown to induce activation of c-Raf-1, but c-Raf 1 has not previously been shown to be activated through the T-cell receptor (TCR) in resting G0 T cells. Using a sensitive immune complex kinase reaction, we show that cross-linking of the stimulatory and costimulatory receptors CD3, CD4, or CD28 induces c-Raf-1 activation in highly purified resting peripheral blood human T cells. In contrast, cross-linking the nonstimulatory receptor CD45 did not induce c-Raf-1. Surprisingly, although earlier studies had shown delayed kinetics in response to Thy-1 stimulation in murine cells, c-Raf-1 activation in response to CD3 cross-linking was one of the earliest measurable events. In spite of its early kinetics, c-Raf-1 activation was found to be downstream of several other early signal transduction events, including activation of a tyrosine kinase and a tyrosine phosphatase. Several lines of evidence suggest that activation of c-Raf 1 in response to TCR stimulation may be PKC-dependent: first, phorbol esters are extremely potent activators of c-Raf-1 in human T cells; second, the kinetics of accumulation of products of phosphatidylinositol hydrolysis coincides with the kinetics of c-Raf-1 activation; and third, physiologic activation of the PLC/PKC pathway through a transfected, G-protein-coupled receptor HM1 induced similar levels of c-Raf-1 activation with a similar time course. We conclude that c-Raf-1 activation is tightly coupled to TCR stimulation and may participate in signal transduction pathways in resting, G0 T cells. The observation that the HM1 receptor can also activate c-Raf-1 suggests that T cells have the capability to utilize both tyrosine kinase-dependent and tyrosine kinase-independent mechanisms of c-Raf-1 activation. PMID- 8409390 TI - Germline and productive C epsilon gene expression during in vivo IgE responses. AB - In vitro studies have established that Ig isotype switching typically involves deletion of CH genes that are located between VDJ and the CH gene that will be expressed, and is preceded by transcription of a germline (g) form of that CH gene. Increases in g epsilon transcript levels are induced by the cytokine IL-4, and always precede switching to IgE. To evaluate whether a similar relationship occurs in vivo, we examined IL-4 mRNA, g epsilon RNA, productive (p) epsilon mRNA, and serum IgE levels in two in vivo systems: one in which the injection of anti-IgD antibody induces mIgD+ B cells to switch to the expression of IgE and to secrete this isotype, and a second in which the injection of anti-IgE antibody stimulates IgE secretion by B cells that had been induced to express membrane IgE by earlier treatment with anti-IgD antibody. Increases in IL-4 transcript levels in anti-IgD-injected mice were followed within 24 h by increases in g epsilon RNA, and, one to two days later, by increased p epsilon mRNA and serum IgE levels. IL-4 antagonists blocked the g epsilon and p epsilon RNA and serum IgE responses in these mice, whereas the injection of otherwise untreated mice with IL-4 stimulated, within 24 h, a large increase in g epsilon RNA levels, followed 1-2 days later by a small increase in p epsilon mRNA. Injection of anti-IgD primed mice with anti-IgE antibody also stimulated increases in IL-4, g epsilon and p epsilon RNA levels; however, the increases in IL-4 and g epsilon RNA were considerably smaller, and the increases in p epsilon mRNA and serum IgE considerably larger, than those observed in anti-IgD antibody-injected mice. IL-4 antagonists blocked the anti-IgE antibody-induced g epsilon RNA response, but not the p epsilon mRNA or serum IgE responses. Thus, IL-4 is required for the induction of g epsilon RNA in at least two in vivo systems, increased g epsilon RNA levels precede increases in p epsilon RNA levels in vivo as in vitro, and neither IL-4 nor g epsilon RNA is required to induce B cells that have already switched to IgE expression to differentiate into IgE-secreting cells. PMID- 8409391 TI - Structural analysis of chicken factor B-like protease and comparison with mammalian complement proteins factor B and C2. AB - Chicken complement factor B-like protease is a glycoprotein of 95 kDa. Activation of chicken serum complement with inulin cleaved the B-like protease into an N terminal Ba fragment of 37 kDa and a C-terminal Bb fragment of 60 kDa. The whole protein and the two fragments were purified by affinity chromatography using mAb to chicken Ba or Bb followed by ion exchange chromatography. Amino acid sequencing showed that chicken B-like protease was cleaved at a site homologous to that cleaved in mammalian complement components B and C2 on activation. Limited tryptic digestion of the B-like protease generated fragments similar to Ba and Bb. More than 200 residues of the Ba sequence and two N-linked glycosylation sites were established by amino acid sequencing of peptides derived by digestion with four proteases. Comparison of human and mouse C2 and B sequences indicated a slower evolutionary rate for B (85% sequence identity) than for C2 (74% sequence identity). Comparison of chicken Ba to human and mouse C2b and Ba showed 42 to 45% sequence identity with respect to C2b fragments, and 46 to 49% sequence identity with respect to Ba fragments. Taking the slower evolutionary rate of factor B into account, chicken factor B-like protease seems to be equally related to mammalian complement components B and C2, and the B-like protease most likely represents the present-day descendant of a common ancestral protein for mammalian B and C2. This conclusion is in agreement with the requirement for the B-like protease in both classical and alternative activation pathways for chicken complement, and with the apparent lack of a chicken serum protein with exclusive C2 activity. PMID- 8409392 TI - Structural analysis of proteolytic products of MHC class II-invariant chain complexes generated in vivo. AB - The MHC class II alpha beta heterodimer associates with invariant (I) chain in the endoplasmic reticulum and remains associated until the complex reaches a post Golgi compartment. During early stages of transport, I chain blocks peptide binding to alpha beta dimers. I chain is proteolytically cleaved in a post-Golgi compartment releasing alpha beta dimers that can bind antigenic peptides and transport them to the cell surface. Human B lymphoblastoid cell lines grown in leupeptin, a sulfhydryl protease inhibitor, accumulate a partial proteolytic product of the I chain called leupeptin-induced protein (LIP). LIP remains associated with alpha beta dimers. We find, using chemical cross-linking, sucrose gradient sedimentation, and size exclusion chromatography, that the alpha beta LIP complex retains the nine-subunit structure described for alpha beta I complexes. Unlike the alpha beta I complex, in certain detergents the alpha beta LIP nonamer is unstable and dissociates into trimers containing one alpha, beta, and LIP molecule. This finding emphasizes the reported stoichiometry of the alpha beta I complex as a nine-subunit structure comprised of three alpha beta I trimers. Also, these data indicate that the region(s) of I chain necessary for retaining the nonameric structure lie within the LIP fragment, but that domains to the C-terminus of the LIP cleavage site act to further stabilize the nine chain structure. In addition, alpha beta I complexes containing forms of human I chain encoding the p35/p43 N-terminal cytoplasmic extension responsible for endoplasmic reticulum retention can transport to post-Golgi proteolytic compartments where LIP is formed. PMID- 8409393 TI - Analysis of Ig H chain gene segment utilization in human fetal liver. Revisiting the "proximal utilization hypothesis". AB - The utilization of Ig H chain gene segments during ontogeny is postulated to result from an immature recombination machinery that preferentially rearranges genes according to chromosomal position. We have tested the "proximal utilization hypothesis" using the two functional members of the VH5 family as a model. Nucleotide sequence analyses of transcripts involving these VH gene segments in 11- to 12-wk fetal liver samples revealed that they were utilized frequently, even though they are 500 kb apart. Although 50% of the H chain rearrangements in our study use the DQ52 gene segment, the most 3' end proximal human D segment, the preference for rearranging D gene segments based on 3' end proximity does not apply to other D segments because different members of the DLR family that are interspersed throughout the D locus are equally rearranged in our sample. The recurrent utilization of the DQ52 and JH4 gene segments and a propensity to preserve the two nucleotides flanking the 3' end of the VH gene segment to generate the amino acid residue at the V-D junction appear to be the major biases in the generation of an otherwise diverse fetal repertoire. PMID- 8409394 TI - Transforming growth factor beta 1 repression of the HLA-DR alpha gene is mediated by conserved proximal promoter elements. AB - Transforming growth factor beta 1 (TGF-beta 1) is a pleiotropic cytokine that decreases the expression of class II MHC Ag in the melanoma cell line Hs294T(c). To investigate the mechanism of this repression, we have examined the effect of TGF-beta 1 on expression of the HLA-DR alpha gene. Both the constitutive level of HLA-DR protein and DR alpha mRNA were repressed by treatment with TGF-beta 1. The proximal 176 bp of the DR alpha promoter were sufficient to confer TGF-beta 1 repression on a reporter gene. Deletional and mutational analysis of the DR alpha promoter revealed that the conserved S and X1 promoter elements were important for basal expression of DR alpha and also mediated the down-regulation by TGF beta 1. Mobility shift assays and in vivo footprinting showed no change in occupancy of the proximal DR alpha promoter after TGF-beta 1 treatment. These results identify the DNA elements that mediate repression of the HLA-DR alpha gene by TGF-beta 1 and suggest that TGF-beta 1 acts at these sites without causing a change in promoter occupancy. PMID- 8409395 TI - Evidence for superantigen production by Yersinia pseudotuberculosis. AB - Yersinia pseudotuberculosis is an enteric pathogen that induces a variety of clinical symptoms, fever, scarlatiniform rash, diarrhea, vomiting, and arthritis. Characteristic histopathologic findings in Y. pseudotuberculosis infection such as lymphoid hyperplasia, typically seen in mesenteric lymph nodes, suggest that the stimulation of a large proportion of T lymphocytes may be involved in the pathogenesis of this infection. In this study, we assessed the mitogenic activity of culture supernatants of the clinical isolates of Y. pseudotuberculosis and investigated the mechanism by which these culture sups activate T cells. The culture sups, as well as partially purified fractions obtained by gel filtration, were found to selectively stimulate T cells bearing V beta 3, V beta 9, V beta 13.1, and V beta 13.2 compared with stimulation by anti-CD3. Furthermore, fibroblasts transfected with different HLA class II molecules, either HLA-DPw9, DQw6, -DR1, or -DR4 Dw15, were capable of presenting Y. pseudotuberculosis culture supernatants to purified T cells. The T cell response to this sup was not restricted by donor HLA-DR types and was not neutralized by antibodies against the known staphylococcal superantigens, Staphylococcal enterotoxin (SE)A, SEB, SEC2, SED, SEE, and TSST1. These results suggest that Y. pseudotuberculosis produces superantigenic toxins that may mediate some of the systemic illnesses associated with infection by this organism. PMID- 8409396 TI - Xid-associated resistance to experimental Chagas' disease is IFN-gamma dependent. AB - In contrast to normal Balb/c, Balb.Xid immunodeficient mice are naturally resistant to Trypanosoma cruzi infection. Thus, Balb.Xid mice control parasitemia, do not show the characteristic wasting in the acute infection and develop no tissue pathology in the skeletal or cardiac muscles in the chronic phase of disease. By in situ hybridization and semiquantitative polymerase chain reaction, the expression of IL genes in spleen cells from Balb/c and Balb.Xid mice were compared after T. cruzi infection. The results showed that Balb.Xid mice produce considerably higher levels of IFN-gamma, IL-2, and IL-4, but lower levels of IL-10, from as early as 4 days after parasite injection. By day 12 of the infection, although IFN-gamma, IL-2, and IL-4 expression was now comparable in both groups, IL-10 levels continue to be lower in Balb.Xid than in control Balb/c animals. The central role of IFN-gamma in the resistance to T. cruzi was confirmed by treatment of Balb.Xid mice with anti-IFN-gamma antibodies that reestablished susceptibility and lead to increased parasitemia and mortality. PMID- 8409397 TI - Tumor-specific lysis of human renal cell carcinomas by tumor-infiltrating lymphocytes. I. HLA-A2-restricted recognition of autologous and allogeneic tumor lines. AB - Metastatic renal cell carcinoma (RCC), like melanoma, belongs to the small group of human tumors in which partial or complete remission has been observed in some patients after treatment with various forms of immunotherapy. In contrast to melanoma, CTL showing MHC-restricted lysis of RCC have not been easily found among tumor-infiltrating lymphocytes (TIL). This has led to the suggestion by some that responses to immunotherapy are mediated predominantly by non-MHC restricted effector cells. We have characterized an MHC-restricted, CD8+ CTL line obtained from an uncloned TIL population of a primary RCC using a low concentration of rIL-2; in fact, these CTL represented a majority of the short term cultured TIL population. The CTL lysed autologous tumor cells but not normal kidney cells or target cells sensitive to non-MHC-restricted effector cells. In contrast, lymphokine-activated killer (LAK) cells grown in a high concentration of rIL-2 from the patient's PBL lysed autologous tumor and normal kidney cells in addition to several allogeneic tumors. The TIL could be expanded optimally using an autologous tumor line retrovirally transduced with the human cDNA encoding IL 2. TIL were 20-fold more potent than LAK cells in eliminating the IL-2 expressing tumor cells in vitro. The cultured TIL utilized a restricted number of V alpha gene families, suggesting that they may recognize only a limited number of MHC peptide complexes presented by autologous tumor cells. HLA-A2 was identified as an MHC restriction molecule for presentation of one tumor-derived peptide to these CTL. Only some allogeneic HLA-A2 RCC tumors were lysed. Sequencing of the second and third exons of the HLA-A2 alleles of these cells revealed that both heterogeneity in MHC and peptide availability influenced CTL recognition. These studies demonstrate that some RCC express common antigenic determinants that can be recognized by MHC-restricted CTL and open the possibility of defining the nature of RCC-derived peptides, which, combined with HLA-A2, can generate specific immune responses. PMID- 8409398 TI - Lymphocyte-vascular adhesion protein-2 is a novel 70-kDa molecule involved in lymphocyte adhesion to vascular endothelium. AB - Extravasation of leukocytes from the blood is essential for normal lymphocyte recirculation as well as in mounting an adequate inflammatory response in different tissues. Leukocyte migration from the blood is controlled by sophisticated interactions between surface receptors on leukocytes and their corresponding endothelial cell ligands. Here we describe a novel adhesion molecule, lymphocyte-vascular adhesion protein-2 (L-VAP-2), recognized by 4G4 mAb that was produced by immunizing mice with an enriched endothelial cell preparation isolated from inflamed human synovium. mAb 4G4 stains a subpopulation of venules in lymphoid and nonlymphoid tissues as well as a few high endothelial venules in lymphoid tissues. L-VAP-2 is constitutively expressed on human umbilical vein endothelial cells, and its expression cannot be up-regulated by cytokines and mitogens such as TNF-alpha, IL-1, and LPS. The Ag is expressed on approximately 20% of PBL, whereas granulocytes and monocytes are negative. On lymphocytes, L-VAP-2 is preferentially expressed on B cells and CD8+ T cells. The molecular mass of L-VAP-2 is approximately 70 kDa as determined from immunoprecipitated, 125I-labeled endothelial cell lysates. The involvement of L VAP-2 in lymphocyte binding to endothelium was tested in vitro using human umbilical vein endothelial cells. Both the intact antibody and F(ab')2 fragments of it consistently inhibited lymphocyte binding to human umbilical vein endothelial cells by approximately 25%. On the basis of the molecular mass estimation and the staining of tissue sections, leukocyte populations, and ICAM-1 and ICAM-2 transfectants, L-VAP-2 appears to be a novel Ag involved in the lymphocyte-endothelial cell interaction. PMID- 8409399 TI - Differential cytokine regulation of complement C3, C4, and factor B synthesis in human intestinal epithelial cell line, Caco-2. AB - In the intestinal tract, the local synthesis of C3 and components of both the classical (C4) and alternative (factor B) C activation pathway has previously been demonstrated in vivo. However, the cellular source of this local C synthesis has not been identified. In this study, we demonstrated the syntheses of C3, C4, and factor B in the human colonic adenocarcinoma cell line Caco-2, which is regarded as a good experimental model of normal human intestinal epithelial cells. The results of metabolic labeling experiments indicated that the intra- and extracellular molecular sizes and subunit structures of Caco-2-derived C3, C4, and factor B were compatible with previously reported values for these components in other cells. The functional activities of C3 and C4 in the supernatants were also demonstrated by hemolytic titration assay. Furthermore, C syntheses in this line were independently upregulated by several human cytokines: C3 synthesis was dose-dependently enhanced by the addition of IL-1 beta or TNF alpha; C4 synthesis was enhanced by the addition of IL-6 or IFN-gamma in the same manner; and the addition of IL-1 beta or IL-6 also induced a dose-dependent increase in factor B synthesis. These enhancing effects were confirmed to be specific for individual cytokines by experiments using anti-human cytokine antibodies. It is likely that intestinal epithelial cells are local production sites of C3, C4, and factor B, and that local C syntheses in the intestine are independently regulated by several cytokines, derived from monocytes/macrophages and T cells resident in the mucosal microenvironment. PMID- 8409400 TI - Antagonistic effects of lipopolysaccharide binding protein and bactericidal/permeability-increasing protein on lipopolysaccharide-induced cytokine release by mononuclear phagocytes. Competition for binding to lipopolysaccharide. AB - Serum proteins play an important role in LPS-induced cell activation. The LPS binding protein (LBP) enhances cellular responses to LPS, whereas the polymorphonuclear leukocyte product bactericidal/permeability-increasing protein (BPI) inhibits LPS-induced cell activation. In this study the influences of LBP and BPI, two proteins with opposite effects, but with considerable sequence homology, on LPS-induced mononuclear phagocytic cell cytokine release was studied. LBP was shown to enhance LPS-induced TNF-alpha, IL-6, and IL-8 release by mononuclear phagocytic cells, whereas BPI inhibited the release of these cytokines. Furthermore, the effects of LBP and BPI on LPS-induced cytokine release by mononuclear phagocytic cells were shown to be counteractive. BPI interfered with the enhancing effect of LBP on the LPS-induced cytokine release. At high LBP to BPI ratios, BPI could no longer inhibit LBP-induced enhancement. In accordance, increasing concentrations of BPI abrogated the LBP effect. Next, it was shown that LBP and BPI compete for binding to LPS by using an assay system that detects binding of free BPI to an anti-BPI mAb. LPS prevented binding of BPI to anti-BPI mAb, whereas preincubation of LPS with LBP prevented the LPS-induced inhibition. Also, it was observed that both BPI and LBP inhibited LPS activity in the chromogenic LAL assay. We conclude from this study that LBP and BPI have counteractive effects on LPS-induced mononuclear phagocytic cell cytokine release by competing for binding to LPS. PMID- 8409401 TI - Particle digestibility is required for induction of the phosphatidylserine recognition mechanism used by murine macrophages to phagocytose apoptotic cells. AB - One of the characteristic features of programmed cell death in vivo is the rapid recognition and removal of apoptotic cells by macrophages. Although there are several potential mechanisms by which the macrophage can identify a cell as apoptotic, it has been shown recently that murine-elicited macrophages stereospecifically recognize phosphatidylserine (PS) exposed on the surface of apoptotic cells. The particulate stimulus, beta-1, 3-glucan, stimulates bone marrow-derived macrophages to express several characteristics of inflammatory macrophages, and induced these cells to recognize PS on apoptotic cells; this activity was correlated with the ability to form rosettes with PS-expressing RBC. Induction of PS recognition in bone marrow-derived macrophages was associated with digestibility of the stimulus, because L, but not D amino acid particles or latex, were able to stimulate macrophage recognition of PS. The requirement for digestibility could be bypassed by the addition of exogenous TGF-beta, which induced macrophage recognition of PS after stimulation with either latex or D amino acid particles. That endogenously produced TGF-beta played a role in the glucan-stimulated response was indicated by the ability of anti-TGF-beta antibodies to inhibit digestible particle-induced recognition of PS. The induction of the PS recognition mechanism correlated well with the expression of other markers for the inflammatory phenotype. These studies indicate that the PS receptor may be a marker for the inflammatory phenotype, which appears to be induced by the phagocytosis of particulate digestible stimuli. Endogenously produced TGF-beta is suggested to play an autocrine or paracrine priming role in the induction of the PS receptor. PMID- 8409402 TI - Overexpression of major heat shock protein hsp70 inhibits tumor necrosis factor induced activation of phospholipase A2. AB - We have recently shown that major heat shock protein (hsp70) protects WEHI-S tumor cells from the cytotoxicity mediated by TNF. In the present study, the mechanism of hsp70-associated TNF resistance was investigated. Overexpression of human hsp70 or inhibition of endogenous hsp70 synthesis by expression of antisense hsp70 RNA did not change the ability of WEHI-S tumor cells to bind TNF or internalize and degrade the receptor-bound TNF. Moreover, TNF-induced activation of NF-kappa B-like transcription factors was unaffected by altered levels of hsp70 as tested by electrophoretic mobility shift assay. Thus, it is unlikely that the resistance is due to changes in TNF receptors or in their ability to transduce signals leading to the regulation of genes, whose expression is regulated by NF-kappa B-like transcription factors. The idea that hsp70 associated TNF resistance is independent of regulation of TNF-induced gene expression was further supported by the results showing that hsp70 protected WEHI S cells from TNF-mediated killing also in the presence of inhibitors of either translation or transcription. Interestingly, TNF-induced activation of arachidonic acid metabolism correlated directly with their sensitivity to TNF and inversely with the amount of hsp70 in the cells. Furthermore, TNF-induced activation of arachidonic acid metabolism was inhibited in WEHI-S cells and two TNF-sensitive human cell lines by induction of the synthesis of endogenous heat shock proteins by heat shock. Even stronger inhibition of arachidonic acid metabolism was seen in WEHI cells rendered TNF-resistant by culturing them in the presence of increasing concentrations of TNF. These cells also had reduced numbers of type 1 TNF receptors. Overexpression of a low molecular weight heat shock protein hsp27 in WEHI-S cells had no effect on any of the parameters studied. These results show that both hsp70-mediated and TNF-induced TNF resistance are associated with a reduced activation of phospholipase A2 suggesting that phospholipase A2 plays an essential role in TNF-mediated cytotoxicity and that hsp70 interferes with the signal transduction pathway leading to its activation. PMID- 8409403 TI - Lipocortin-1 fragments inhibit neutrophil accumulation and neutrophil-dependent edema in the mouse. A qualitative comparison with an anti-CD11b monoclonal antibody. AB - The activity of the steroid-inducible protein lipocortin-1 (LC1; with a primary sequence of 346 amino acids; also called annexin 1), a fragment corresponding to amino acids 1-188 and a short peptide from the N-terminus (amino acid 2-26) were tested for anti-inflammatory actions in three models of acute inflammation in the mouse in comparison with a mAb anti-CD11b (alpha CD11b). In the mouse air-pouch model LC1, fragment 1-188 and peptide Ac2-26 exhibited powerful inhibitory effects (ED50 approximately 5.2, 38 and 88 micrograms/mouse, respectively) on leukocyte migration elicited by IL-1. LC1 was approximately 200 times more potent than Ac2-26 on a molar basis although both gave maximal inhibitions, in contrast fragment 1-188 only produced a partial dose-response curve. LC1 was approximately 20 times more potent on a molar basis in this assay than the alpha CD11b mAb. Peptide Ac2-26 and the mAb alpha CD11b also blocked cell migration into the air pouch induced by IL-8 with approximately the same potency. In the mouse skin edema and zymosan peritonitis assays Ac2-26 was inhibitory (ED50 of 200 micrograms/mouse) but less so than the alpha CD11b antibody (ED50 approximately 0.5 mg/mouse). Both LC1 (10 micrograms) and Ac2-26 (200 micrograms) completely blocked FMLP-induced neutropenia in the mouse. Studies using an inactivated LC1 preparation, which binds to the same high affinity binding sites as the biologically active material, indicated that the short peptide acts on the same sites as the native LC1. This study confirms the activity of LC1 in another model of experimental inflammation and suggests that it acts partly through inhibition of leukocyte activation with an overall effect qualitatively comparable to the blocking of CD11b portion of a beta 2-integrin complex. It also shows that peptides derived from the N-terminal domain of LC1 may mimic the activity of the full length molecule and points the way for a new family of anti-inflammatory substances that inhibit leukocyte trafficking. PMID- 8409404 TI - C1Q, a subunit of the first component of complement, enhances the binding of aggregated IgG to rat renal mesangial cells. AB - Previous reports have shown the presence of C1Q-R on monocytes, macrophages, polymorphonuclear cells, fibroblasts, platelets, lymphocytes, and endothelial cells. The present study demonstrates a functional C1Q-R on rat renal mesangial cells (MC). Incubation of MC with increasing concentrations of [125I]C1Q resulted in a dose-dependent binding of [125I]C1Q to MC; the binding of [125I]C1Q was inhibitable by excess unlabeled C1Q or C1Q stalks whereas BSA and C1Q globular heads had no effect. Scatchard analysis of the data revealed the presence of 6.2 x 10(7) binding sites/cell with an affinity of 4.9 x 10(6) M-1 for C1Q. Immunoprecipitation of 125I-labeled MC membrane proteins with C1Q or mAb directed against human C1Q-R revealed a single 66- to 68-kDa band under reducing conditions. We have shown previously that soluble stable aggregates of IgG bind to rat MC in a dose-dependent fashion. In addition the presence of a receptor for IgG has been described on rat MC. In order to find out whether there is a cooperative effect between C1Q and AlgG in binding of [125I]AlgG to MC, we incubated [125I]AlgG in the presence of increasing concentrations of C1Q, and showed a 5- to 15-fold enhancement of binding of [125I]AlgG to MC. Neither heat inactivated C1Q nor C1Q stalks were able to enhance the binding of [125I]AlgG to MC. Enhanced binding by C1Q was only observed when aggregated IgG was used; the binding of monomeric IgG to MC was not affected by C1Q. These studies indicate that there is a cooperative effect between Fc gamma R and C1Q-R on MC in the recognition of immune complexes. PMID- 8409405 TI - Macrophage inflammatory protein-1 alpha rapidly modulates its receptors and inhibits the anti-CD3 mAb-mediated proliferation of T lymphocytes. AB - Macrophage inflammatory protein-1 alpha (MIP-1 alpha) is a member of the intercrine/chemokine family which consists of basic, heparin-binding, small molecular weight proteins. We have previously shown that a T cell line, CTLL-R8, carried high-affinity receptors for MIP-1 alpha and the proliferation of CTLL-R8 cells was inhibited by murine recombinant (mr) MIP-1 alpha. We extended our previous studies to murine resting splenic T lymphocytes to determine whether the inhibition of T cell proliferation is a general property of MIP-1 alpha. The resting splenic T cells carried approximately 680 high-affinity binding sites for mrMIP-1 alpha; more than 90% of the primary T cells carried MIP-1 alpha receptors. When the T cells were stimulated with immobilized anti-CD3 mAb in the presence of accessory cells, the MIP-1 alpha binding was reduced. The lowest binding was obtained 2 h after anti-CD3 mAb stimulation due to the internalization of MIP-1 alpha receptors. mrMIP-1 alpha inhibited the anti-CD3 mAb-mediated proliferation of murine splenic T lymphocytes. The maximum inhibition was obtained when mrMIP-1 alpha was added 30 min before anti-CD3 mAb stimulation. Slight inhibition of T cell proliferation was observed when mrMIP-1 alpha was added at the same time as anti-CD3 mAb stimulation. These results indicate that T lymphocytes are regulated negatively by MIP-1 alpha, which occurs when the T cells are exposed to MIP-1 alpha before activation. The negative effect of MIP-1 alpha seems to be mediated in part by the inhibition of IL-2 production, for there was a reduction in both the IL-2 mRNA levels and the IL-2 activity in supernatants from T cells preincubated with MIP-1 alpha before anti CD3 mAb stimulation. PMID- 8409406 TI - Suppression of monocyte function and differential regulation of IL-1 and IL-1ra by IL-4 contribute to resolution of experimental arthritis. AB - IL-4 has diverse effects on hematopoietic cells, including the ability to suppress certain mononuclear cell functions. To evaluate the effect of IL-4 on the evolution of acute and chronic arthritis, murine recombinant IL-4 was administered systemically to animals receiving an arthropathic dose of group A streptococcal cell wall fragments. Daily treatment with IL-4 had a minimal effect on the acute phase, but significantly suppressed the chronic, destructive phase. By 4 wk after initiation of disease, the articular index of IL-4-treated animals was reduced > 60% (articular index = 4 +/- 0.9) compared with the untreated rats (11.5 +/- 0.48, p < 0.001). A substantial decrease in the influx of inflammatory cells and virtual elimination of pannus and erosions occurred after IL-4 therapy. Associated with the reduced accumulation of mononuclear leukocytes was a decrease in their proinflammatory functions including cytokine production and reactive oxygen intermediate metabolism. These observations are consistent with the selective effects of IL-4 on phagocytic cell function demonstrated in vitro. Furthermore, IL-4 induced gene expression for IL-1ra, a protein that antagonizes the action of IL-1 by binding to the IL-1 receptor without agonist activity. Through an expanding spectrum of effects on monocyte-macrophage phenotypic and functional parameters, IL-4 is emerging as an important inhibitor of cell mediated immune responses and pathogenic processes. PMID- 8409407 TI - Clonal characterization of the human IgG antibody repertoire to Haemophilus influenzae type b polysaccharide. V. In vivo expression of individual antibody clones is dependent on Ig CH haplotypes and the categories of antigen. AB - Antibodies (Ab) to the polysaccharide capsule of Haemophilus influenzae type b (Hib-PS) provide protection against Haemophilus influenzae type b disease in children, and Hib-PS vaccines with different immunologic properties are widely used clinically. The repertoire of human anti-Hib-PS Ab induced by these vaccines is relatively restricted and can be divided into two types by the structure of the light chain V region. Ab using A2-V kappa II gene product, which account for the majority of anti-Hib-PS Ab response in most patients, show little somatic mutations. In contrast, non-Ab using A2-V kappa II gene product use VL genes from the V kappa I, V kappa II, V kappa III, V kappa IV, and V lambda subgroups, are variably expressed among patients, and contain somatic mutations. To further study the expression of these two types of anti-Hib-PS Ab, we have produced KB13, a mAb specific for V kappa II subgroup, and used mAb specific for various other VL subgroups to develop immunoassays specific for anti-Hib-PS Ab of each VL subgroup. When Ig allotypes were studied for the effect on the Ab repertoire, A2 V kappa II (A2) Ab were found to be expressed less in patients expressing fb or zag CH haplotypes (p < 0.05). When the T cell-independent Hib-PS carbohydrate vaccine was compared to two T cell-dependent Hib-PS protein conjugate vaccines for their effect on Ab repertoire, Ab using V kappa III VL were found to be more often elicited with the conjugate vaccines than with the Hib-PS carbohydrate vaccine (p < 0.01). Thus, individual members of the anti-Hib-PS Ab repertoire differ not only in their V region structure but also in the control of their expression. PMID- 8409408 TI - Prevention of autoimmune type I diabetes by CD4+ suppressor T cells in superantigen-treated non-obese diabetic mice. AB - The development of autoimmune type I diabetes in the NOD mouse appears to be controlled by both genetic and environmental factors. This investigation was initiated to determine whether exogenous superantigens, as environmental factors, can influence the development of diabetes. Several staphylococcal enterotoxins (SE) (SEA, SEC1, SEC2, or SEC3), which are known superantigens, were injected i.v. into female NOD mice at 4 and 10 wk of age. At 32 wk of age, the incidence of diabetes in the SE-treated mice ranged from 6 to 12.5%; this was significantly lower than that of mice treated with PBS--64%. There was no significant difference in effectiveness among the various SE used. SE induced a modest decrease in T lymphocytes bearing specific V beta TCR 2 wk after injection, but this effect did not persist past 4 wk. To elucidate the mechanism of the SE effect, suppressor activity in SE-treated mice was evaluated. Splenocytes from SE treated mice inhibited the transfer of diabetes by splenocytes from acutely diabetic NOD mice when injected into irradiated young NOD mice; only 10% became diabetic. In contrast, 83% of the mice receiving splenocytes from PBS-treated control mice became diabetic. Suppressor activity of splenocytes from SE-treated mice was diminished by the depletion of CD4+ T cells, but not by the depletion of CD8+ T cells, indicating that the suppressor cells belonged to the CD4+ T class of lymphocytes. On the basis of these observations, we conclude that exogenous superantigens activate CD4+ suppressor T cells, leading to the prevention of autoimmune type I diabetes in NOD mice. PMID- 8409409 TI - T cells of patients with the Wiskott-Aldrich syndrome have a restricted defect in proliferative responses. AB - The Wiskott-Aldrich syndrome (WAS) is a disease of profound thrombocytopenia and severe immune defects caused by an unidentified defective X chromosome gene. In this study, T lymphocyte function is examined using a panel of allospecific WAS patient T cell lines, previously found to express the abnormal disease gene and the cytoarchitectural defect characteristic of the disease. Although T cell lines from normal individuals proliferate vigorously in response to immobilized anti CD3 mAb OKT3 and SPV-T3b, five of seven WAS patient T cell lines failed to proliferate and two lines showed significantly decreased proliferation when challenged with the immobilized anti-CD3 mAb. The deficient responsiveness of the WAS T cell lines to immobilized anti-CD3 mAb is a restricted defect, because the cells proliferate normally when challenged with allospecific Ag, PHA, or PMA plus ionomycin. Addition of anti-CD28 mAb did not correct the deficient proliferation of the WAS cells challenged with immobilized anti-CD3. Deficient response of the WAS T cell lines to immobilized anti-CD3 was detected also when earlier events of the proliferation process, IL-2 production and up-regulation of activation Ag CD69 and CD28, were measured. On the other hand, WAS cell lines did not differ from normal cell lines in binding of anti-CD3 mAb, mobilization of Ca2+ in response to soluble OKT3, and tyrosine phosphorylation and GTP binding of the CD3 zeta-chain in response to OKT3. Cumulatively, these findings demonstrate a striking restricted defect in the proliferative response of WAS T cells, which because it is found in cell lines free of secondary changes that occur in the patient circulation must be a reflection of the inherited defective disease gene product. PMID- 8409410 TI - Superantigenic properties of a novel mitogenic substance produced by Yersinia pseudotuberculosis isolated from patients manifesting acute and systemic symptoms. AB - A novel product from Yersinia pseudotuberculosis exhibiting mitogenic activity for human PBMC (designated Y. pseudotuberculosis-derived mitogen, YPM) was examined for its biologic activities for human lymphocytes. YPM induced a substantial proliferative response and IL-2 production at 0.1 ng/ml or more in whole PBMC but not in T-depleted PBMC. IL-2 production occurred within 12 h after YPM stimulation. T cells from PBMC produced IL-2 in the presence of L cells transfected with HLA DR genes or DP genes but not in the presence of control L cells used as accessory cells. Paraformaldehyde fixation did not abolish the AC activity of the DR+ L cells. The results suggest that YPM bind directly to HLA class II molecules. Analysis of the TCR V beta element using the polymerase chain reaction revealed that YPM selectively activated human T cells bearing V beta 3, V beta 9, V beta 13.1, and V beta 13.2 in TCR. These results indicate that YPM is a potent T cell activator and has superantigenic properties (abilities to bind directly to MHC class II molecules and selectively stimulate T cell populations bearing particular V beta elements in TCR). The role of YPM in the mechanism of pathogenesis of Y. pseudotuberculosis infections manifesting acute and systemic clinical symptoms is discussed. PMID- 8409411 TI - Peripheral B cell maturation. II. Heat-stable antigen(hi) splenic B cells are an immature developmental intermediate in the production of long-lived marrow derived B cells. AB - HSAhi B cells comprise 5 to 10% of adult mouse splenic B cells and are phenotypically and functionally immature. To assess their origin and relationship to mature, heat-stable Ag (HSA)lo B cells, we determined HSA and surface IgD phenotype among splenic B cells throughout development, as well as during reconstitution of lethally irradiated adults given adult B-depleted bone marrow. In each case, HSAhi splenic B cells predominate during the earliest stages of B cell genesis. Furthermore, 5-bromo-2'-deoxyuridine labeling experiments indicate rapid turnover within both the marrow and peripheral HSAhi pools, and adoptive transfer studies show that peripheral HSAhi splenic B cells differentiate to HSAlo within 4 days. Finally, splenic HSAhi B cells reconstitute both primary and memory humoral responses. Together, these data indicate that splenic B cells in the HSAhi subset are an intermediate maturational stage in the adult periphery. PMID- 8409412 TI - Human cytomegalovirus down-regulates HLA class I expression by reducing the stability of class I H chains. AB - Human CMV (HCMV) infection leads to an almost complete inhibition of expression of MHC class I proteins. After infection with HCMV, the biosynthesis of HLA class I molecules was examined in human lung fibroblasts and in mouse fibroblasts transfected with genes encoding the human class I H chain HLA-B27 and human beta 2-microglobulin (beta 2m). In both cell types, we observed a large decrease in steady state levels specific for human class I H chains--both free H chains and those complexed with beta 2m. In the mouse cells transfected with HLA class I, infection did not affect levels and assembly of mouse class I H chains with human beta 2m. The effect of HCMV infection on class I expression is insensitive to phosphonoacetic acid, suggesting the involvement of immediate early or early viral proteins. Pulse-chase analysis showed that the low steady state level of class I H chains in HCMV-infected cells is not the result of a reduced rate of synthesis. Rather, we observed accelerated degradation of class I H chains, regardless of their association with beta 2m. We conclude that HCMV reduces human MHC class I protein levels by interference with the stability of class I H chains. PMID- 8409413 TI - Most gamma delta T cells develop normally in the absence of MHC class II molecules. AB - MHC class I and class II molecules are essential for intrathymic maturation of a normal repertoire of alpha beta T cells. The development of gamma delta T cells may similarly require exposure to conventional MHC or MHC-like molecules for appropriate negative and positive selection. The availability of mice that are devoid of cell surface expression of MHC class II molecules allowed us to test directly the hypothesis that conventional class II molecules play a role in the development of gamma delta T cells. The proportions of gamma delta cells in thymus, lymph nodes, and spleens were indistinguishable between class II deficient mice and littermate controls by FACS analysis. gamma delta T cells from class II-deficient mice proliferated normally in response to anti-TCR mAb. Examination of epidermal sheets from ear, torso, and tail skin demonstrated that class II-deficient mice had the same population density and distribution of gamma delta+ dendritic epidermal T cells as that of control littermates. gamma delta Cells in the epidermis of class II-deficient mice expressed Thy-1 and CD3 and were predominantly V gamma 3+. There were no significant differences in the immunophenotype of class II-deficient mice and their normal littermates in two color immunofluorescence analysis of intestinal intraepithelial lymphocytes. Class II-deficient mice and control mice had 29 to 39% gamma delta bearing intestinal intraepithelial lymphocytes, of which 16 to 21% expressed V delta 4. Dendritic cells that stained positively for Thy-1, CD3, and gamma delta were identified in the vaginal epithelium of class II-deficient mice and their normal littermates. Our results indicate that MHC class II expression is not essential for the development of most gamma delta T cells. PMID- 8409414 TI - B cell antigen receptor cross-linking triggers rapid protein kinase C independent activation of p21ras1. AB - Membrane Ig (mIg) functions in binding and internalization of Ag for subsequent processing and presentation to T cells, as well as in transmembrane transduction of signals that lead to cell activation, proliferation, and differentiation. Tyrosine kinase activation and subsequent phosphatidylinositol hydrolysis and Ca2+ mobilization are clearly important intermediary events in receptor-mediated B cell activation. However, many details of the cellular signal transduction pathways utilized by this receptor are not resolved. Recent studies that demonstrated co-capping of mIg and the proto-oncoprotein p21ras suggested that this low m.w. GTP-binding protein may function in mIg-mediated signal transduction. p21ras has been implicated in some but not all protein tyrosine kinase/phospholipase C involving signaling pathways. To explore the potential role of p21ras in B cell Ag receptor-mediated signaling, we assessed the effect of Ag receptor ligation on the proportion of p21ras in the active GTP-bound state. We present evidence that p21ras is activated by mIgM and mIgG cross linking by anti-receptor antibodies as well as by Ag. Depending upon the stimulus employed, this response is detectable within 1 min and occurs with similar kinetics as inductive protein tyrosine phosphorylation and Ca2+ mobilization. Ag dose dependence of this response is similar to that of inductive protein tyrosine phosphorylation. In these cells p21ras is also activated by PMA suggesting that p21ras activation after receptor cross-linking may be mediated by an effector molecule that functions downstream from protein kinase C (PKC). However, the kinetics of p21ras activation after mIg cross-linking are inconsistent with the possibility that PKC functions as the sole mediator of p21ras activation in this system. Finally, under conditions in which the PKC inhibitor calphostin C blocks PMA-induced p21ras activation, it does not inhibit Ag-induced p21ras activation. These data suggest that PKC effector mechanisms play a negligible role in p21ras activation during mIg-mediated signaling. PMID- 8409415 TI - Evidence that murine V gamma 5 and V gamma 6 gamma delta-TCR+ lymphocytes are derived from a common distinct lineage. AB - Murine V gamma 5 and V gamma 6+ T lymphocytes develop sequentially in the thymus during fetal and newborn life, giving rise to intraepithelial lymphocytes of the epidermis and female reproductive tract. In analyzing a panel of gamma delta T cell hybridomas derived from various tissues, we found that all V gamma 6+ cells tested (n = 25) expressed V gamma 5-C gamma 1 mRNA, whereas none of a panel of V gamma 6- cells analyzed (n = 33) expressed this mRNA. V gamma 6 mRNA was rare in V gamma 5+ cells (1 of 9), and absent in all other gamma delta T cells (n = 10). These findings suggest that most, if not all, V gamma 6-TCR+ lymphocytes arise from precursor cells that have previously rearranged the V gamma 5 gene, and indicate that V gamma 5 and V gamma 6+ lymphocytes belong to a common, distinct lineage that does not give rise to other gamma delta T cells. PMID- 8409416 TI - Sequential switching from mu to epsilon via gamma 4 in human B cells stimulated with IL-4 and hydrocortisone. AB - The molecular events leading to IgE synthesis in human B cells stimulated with IL 4 and hydrocortisone were analyzed. IL-4, but not hydrocortisone, induced C epsilon germ line transcription. However, hydrocortisone increased the levels of IL-4-induced germ line C epsilon transcripts by twofold and delivered the signal required for transcription of mature C epsilon mRNA. Nested primer polymerase chain reaction of high m.w. DNA revealed deletional switch recombination occurring in B cells sorted for lack of expression of surface IgE and stimulated with both IL-4 and hydrocortisone, but not in B cells stimulated with IL-4 alone or hydrocortisone alone. DNA sequence analysis of 10 switch fragments revealed direct joining of S mu to S epsilon in eight fragments, one of which exhibited an 876-bp deletion in S mu. The ninth fragment contained a 50-bp insertion at the S mu/S epsilon junction, which was likely to be derived from S gamma 4. The sequence of the 10th fragment was consistent with either a 17-bp insertion at the S mu/S epsilon junction derived from S gamma 4 or with a complex 38-bp deletion within S epsilon. Mapping of the switch junction sites showed "hot spots" for recombination within S mu but not within S epsilon. These findings indicate that hydrocortisone induces S mu-S epsilon deletional switch recombination in IL-4 treated B cells, and support a model of sequential isotype switching from IgM to IgE via IgG4. PMID- 8409417 TI - TGF-beta is a bidirectional modulator of cytokine receptor expression on murine bone marrow cells. Differential effects of TGF-beta 1 and TGF-beta 3. AB - Transforming growth factor beta (TGF-beta), an immunomodulator, has inhibitory as well as stimulatory effects on bone marrow cells. In this study, we demonstrate that TGF-beta 1 also is a bidirectional modulator of CSF receptor expression on murine bone marrow cells. TGF-beta 1 up-regulated granulocyte-macrophage (GM)-CSF receptor expression in a time- and dose-dependent manner, with a maximum up regulation of 64% by 48 h at 20 ng/ml. In contrast, TGF-beta 1 down-modulated IL 3 and CSF-1 receptor expression by 54 and 55%, respectively, by 24 h. TGF-beta 1 did not affect G-CSF receptor expression, in agreement with its inability to affect G-CSF-induced proliferation. The CSF receptor modulation induced by TGF beta 1 preceded its effects on CSF-stimulated proliferation. The effects of TGF beta on CSF receptor expression were isoform dependent, thus TGF-beta 3 was a 10 fold more potent inhibitor of both IL-3-induced colony formation and IL-3 receptor expression than TGF-beta 1, whereas TGF-beta 1 was a more potent stimulator of GM-CSF-stimulated colonies and GM-CSF receptor expression than TGF beta 3. Therefore, the ability of TGF-beta to modulate the CSF receptor density/cell and/or the actual number of progenitors expressing CSF receptors directly correlates with the multifunctional effects of TGF-beta in hematopoiesis. PMID- 8409418 TI - Synthesis of T helper 2-type cytokines at the maternal-fetal interface. AB - Clinical and experimental evidence has indicated that the maternal immune response is biased toward antibody production and away from cell-mediated immunity during pregnancy, especially in the vicinity of the fetoplacental unit. Because antibody responses are often associated with the Th2 cytokine pattern, this suggests that Th2-type cytokines might predominate locally in the regulation of the maternal immune response. In order to test this hypothesis, we examined the local and distal release of cytokines during murine pregnancy using ELISA assays. We report here that the Th2-specific cytokines IL-4, IL-5, and IL-10 were readily detectable in cell supernatants derived from fetal-placental units in all three trimesters of gestation. IL-3 was also present. These cytokines were detected in lysates of freshly isolated, day 12 decidual and placental cells, and in supernatants as early as 15 min after the beginning of culture. The presence of functional IL-10 was confirmed by specific bioassay. IL-10 mRNA was localized to the decidua at day 6 of gestation by in situ hybridization. IFN-gamma was also found in the supernatants from the first trimester of pregnancy, but was barely detectable in the second, and undetectable in the third trimester. Cytokine expression was consistently detected in samples from individual mice. None of these cytokines was produced by unstimulated spleen or mesenteric lymph nodes from pregnant mice. IL-4, IL-10, and IFN-gamma were produced by Con A-stimulated spleen cells from virgin mice, but in ratios opposite to those found in the placenta. These observations indicate that Th2-specific cytokines are normally produced at the maternal-fetal interface. The continuous presence of IL-4, IL-5, and IL-10, with early and transient expression of IFN-gamma, can provide a molecular basis for the antibody/Th2-like bias of the maternal immune response during pregnancy. PMID- 8409419 TI - Normal sequence of phenotypic transitions in one cohort of 5-bromo-2' deoxyuridine-pulse-labeled thymocytes. Correlation with T cell receptor expression. AB - "In vivo" kinetics of T cell differentiation and TCR expression in the normal murine thymus were re-evaluated using a new technique for simultaneous detection of bromodeoxyuridine and two surface markers. The transition from CD4-8- precursors to CD4+8+ immature cells was directly observed during cell proliferation, and shown to proceed through transitory intermediates expressing no or low amounts of CD4. CD3-TCR expression also started during this transition and resulted in the production of a majority of TCRlo cells but also of a significant number (1 to 2 x 10(6) of TCRhi immature (heat-stable Ag+) thymocytes. After cessation of proliferation, the maturational transition from CD4+8+ to CD4+8- and CD4-8+ (in this order) was restricted to TCRhi cells produced during CD4+8+ cell generation. The acquisition of the single positive phenotype preceded HSA down-regulation, suggesting that maturation of TCRhi thymocytes proceeds in two separate steps. The major TCRloCD4+8+ subset appeared a dead end subset and showed no up-regulation of TCR expression at any time. PMID- 8409420 TI - Antigen presentation by B lymphoma cells. Requirements for processing of exogenous antigen internalized through transferrin receptors. AB - The Ag, pigeon cytochrome c, was delivered to the early endosomes in a form coupled to human ferric transferrin that entered APC through transferrin receptors. The processing of the transferrin-Ag conjugate by B lymphoma cells was compared with that of unconjugated native Ag that entered APC by a nonreceptor mediated mechanism. Within 5 min after internalization, catabolized conjugate was detected in isolated early endosomes and did not accumulate in these organelles. Analysis of the rapid catabolism of the conjugate demonstrated that the Ag, not the transferrin, portion of the molecule was degraded by the APC, suggesting that similar proteases may mediate the processing of the conjugate and native Ag. The processing mechanisms of these molecules shared similarities. Treatment of APC with chloroquine or paraformaldehyde interfered with the stimulation of Ag specific CD4+ T cells by both transferrin-Ag conjugate and native Ag. However, the T cell responses to the conjugate and native Ag were different in two important respects. First, T cell activation by the conjugate began at an earlier time point and occurred at a faster rate than T cell stimulation by the same concentration of native Ag during a 3-h time course. Second, the T cell response to the conjugate, but not to native Ag, was diminished by treating APC with cycloheximide, a reversible protein synthesis inhibitor. This partial inhibition of the conjugate response by cycloheximide could not be attributed to significant effects on transferrin receptor expression, or on internalization, recycling, or degradation of the conjugate. The differential cycloheximide-sensitivity of the T cell responses indicates that the processing pathways of the two molecules are different. Our findings suggest that the early endosomes may function as an Ag processing compartment, and that more than one pathway may lead to productive processing in B lymphoma cells. PMID- 8409421 TI - The functional basis of minor histocompatibility loci. AB - This work addresses the functional basis of classical minor histocompatibility (H) loci. We focus on the H-3 locus, which is actually a complex genetic unit to which the phenotypic trait of tissue rejection, genes whose products stimulate specific subsets of T cells, and Ir genes have been mapped. To clarify how these genes relate to one another and to the trait of tissue rejection, strains of intra-H-3 recombinant mice were produced and analyzed. These mice allowed us to selectively elicit immune responses to Ag (referred to as type I Ag) that stimulate MHC class I-restricted CTL, or Ag (referred to as type II Ag) that stimulate MHC class II-restricted Th. The splitting of H-3 in this manner resulted in a dramatic diminution of the skin allograft response, and with rare exception, an elimination of the CTL response after spleen cell immunization. A selective response to type I Ag resulted in slow, incomplete skin allograft rejection that demonstrated both CD4+ cell-dependent and -independent components. A selective response to the type II Ag failed to result in allograft rejection. The type II Ag did, however, act as an Ir gene that determined whether responses to type I Ag could occur. Altogether, the results indicate that the trait of tissue rejection associated with H-3 is a consequence of the strongly synergistic effects of Th-CTL collaboration induced by products of type I and type II genes. Moreover, the results suggest a genetic explanation for some of the Ir gene effects associated with H-3. PMID- 8409422 TI - Characterization of a cell surface glycoprotein IPO-3, expressed on activated human B and T lymphocytes. AB - We characterize the expression, biochemical structure, and function of a novel glycoprotein, IPO-3, up-regulated on activated human lymphocytes. IPO-3 is found on activated B cells, B cell lines, and hairy cell leukemias but is not expressed on T cell or nonlymphoid cell lines. IPO-3 is not B cell-specific as it is detected at low levels on CD45RO+ CD45RA- peripheral blood T cells and CD4+CD8+CD45RO+ CD45RA- thymocytes. The IPO-3 Ag is a single-chain heavily N glycosylated phosphoglycoprotein approximately 75 to 95 kDa in size with a 42-kDa protein core. In vitro kinase assays revealed that IPO-3 has a protein kinase activity associated with it that is maintained even in Nonidet P-40 lysates. IPO 3 is up-regulated on resting B cells within 16 h after activation with different signals including anti-IgM, IL-4, or mAb to CD40, CD20, or Bgp95. It could also be induced on T cells via CD3-cross-linking, but the kinetics of IPO-3 induction was slower on T cells than on B cells. Cross-linking IPO-3 on B cells with mAb did not induce proliferation alone but did augment proliferation promoted by IL-4 and anti-CD40 and did trigger increases in [Ca2+]i in resting B cells. Binding of IPO-3 could not be inhibited by a variety of mAb to previously identified activation markers. Thus, the IPO-3 glycoprotein appears to be a novel marker of activated B and T lymphocytes, which may play a role in the regulation of lymphocyte activation. PMID- 8409423 TI - Transforming growth factor-beta 1 is required for secretion of IgG of all subclasses by LPS-activated murine B cells in vitro. AB - Addition of transforming growth factor-beta 1 (TGF-beta 1) to in vitro cultures of murine B cells activated with bacterial LPS selectively stimulates IgG2b and IgA class switching and decreases cellular proliferation. To assess a possible role for endogenous TGF-beta in modulating the Ig isotypes produced by LPS activated cells, we utilized a neutralizing anti-TGF-beta mAb to abrogate endogenous TGF-beta activity. Anti-TGF-beta antibody, over a range of relatively low cell densities, strikingly inhibited both IgG3 and IgG2b production in response to LPS, with little or no change in the concentrations of secreted IgM. This effect of anti-TGF-beta antibody was specific, since it did not occur with an isotype-matched control mAb and was completely reversed with exogenous TGF beta 1. Optimal IgG3 secretion occurred at concentrations of TGF-beta that were approximately eightfold lower than that necessary for maximal synthesis of IgG2b. Neutralization of endogenous TGF-beta in LPS-activated cultures was associated with an approximately twofold increase in proliferation and viable cell yields, a modest decrease in the percentage of membrane (m)IgG2b+ cells, and a modest increase in the percentage of mIgG3+ cells. This latter finding indicated that TGF-beta was not required for IgG3 class switching, but for maturation of mIgG3+ cells into Ig secretors. Highly purified B cells, obtained by electronic cell sorting, released active TGF-beta in response to LPS and showed a similar marked reduction in LPS-mediated IgG3 and IgG2b secretion in the presence of anti-TGF beta antibody. Abrogation of endogenous TGF-beta activity in LPS-activated cultures also resulted in a striking reduction in IFN-gamma-mediated IgG2a production, and a more modest decrease in the synthesis of IgG1 and IgE in the presence of IL-4. These data indicate that relatively low concentrations of TGF beta are essential for stimulating optimal IgG secretion by LPS-activated B cells, in an Ig isotype-nonspecific manner, and may regulate these responses in an autocrine fashion. PMID- 8409424 TI - Stimulation of human T-cell proliferation by specific activation of the 75-kDa tumor necrosis factor receptor. AB - TNF-alpha can enhance the proliferation of human thymocytes stimulated by the comitogen Con A. To determine which of the two different TNF receptors is responsible for signaling this cellular response, we investigated the proliferation of human thymocytes in response to agonistic antibodies specific for the two TNF receptor types. In contrast to previously examined TNF activities in human cells, thymocyte proliferation was stimulated in response to rabbit polyclonal antibodies directed against the 75-kDa TNF receptor (TNF-R2), but not those directed against the 55-kDa TNF receptor (TNF-R1). Lymphotoxin (TNF-beta) was also shown to stimulate human thymocyte proliferation, demonstrating that TNF beta can initiate a biologic response that is mediated by TNF-R2. TNF-R2-mediated T-cell proliferation was not restricted to the immature T cells within the thymus, as the anti-TNF-R2 antibodies also stimulated the proliferation of peripheral T cells. As a first step toward identifying a specific agonist of TNF R2 with therapeutic potential, 10 anti-TNF-R2 mAb were examined for potential agonist activity. Nine of these significantly stimulated human thymocyte proliferation with maximal responses ranging from twofold to significantly greater than that obtained with TNF-alpha by itself. PMID- 8409425 TI - Unexpected lack of reactivity of allogeneic anti-Ia monoclonal antibodies with MHC class II molecules expressed by mouse intestinal epithelial cells. AB - We have examined MHC class II molecules expression by murine gut epithelial cells using a large panel of anti-Ia antibodies. In contrast to conventional APC (i.e., B cells and macrophages), only two anti-Ia antibodies reacted with enterocytes: a xenogeneic rat anti-Ia mAb (CD311) directed against a monomorphic class II determinant, and a polyclonal antiserum directed against both I-A and I-E heterodimers. In contrast, allogeneic anti-Ia mAb were either unreactive (17 of 20) or reacted weakly (3 of 20) with enterocytes, even after in vivo treatment with IFN-gamma. This pattern of Ia reactivity of epithelial cells was tissue specific (restricted to gut mucosa) and cell specific (restricted to gut epithelial cells). Biochemical and molecular studies confirmed that enterocytes expressed I-A and I-E isotypes on their cell surface and contained mRNA of both subregion loci. Interestingly, enterocytes appeared deficient in expression of the MHC class II-associated invariant chain, and are not able to stimulate allogeneic T cells. These data suggest that gut epithelial cells express a conformation of class II molecules, antigenically distinct from that expressed on conventional APC. PMID- 8409426 TI - Construction of a human Ig combinatorial library from genomic V segments and synthetic CDR3 fragments. AB - A naive combinatorial Ig library was constructed from semi-synthetic V genes consisting of human genomic V segments and synthetic CDR3 fragments. VH and V kappa segments were amplified from human genomic DNA by polymerase chain reaction using V subgroup-specific primers. The amplified VH and V kappa segments were combined with synthetic oligonucleotides containing a J region and CDR3 with amino acid sequence variations, resulting in complete V genes. These V genes were cloned into a phagemid expression vector in a single-chain form fused to the carboxyl-terminus of the M13 minor coat protein III. Phagemid particles displaying the single chain hybrid proteins on their surface were screened with Con A as Ag. Several clones showing specific binding to Con A were obtained after four rounds of selection and were further analyzed for their binding properties and DNA sequences. This method provides a novel way to create a naive combinatorial library without using mRNA from B lymphocytes as template. The method should be useful to isolate human antibodies that react with self-Ag. PMID- 8409427 TI - Independently derived IgG anti-DNA autoantibodies from two lupus-prone mouse strains express a VH gene that is not present in most murine strains. AB - The origin and structure of two clonally unrelated IgG anti-DNA autoantibodies from lupus-prone MRL/Ipr mice were examined. One of these antibodies, H241, binds dsDNA and glomeruli and deposits in the kidneys of normal mice, whereas the other, H102, binds only ssDNA and does not deposit in kidneys. The VH genes of these two antibodies were almost identical to each other and were frequently expressed in anti-DNA antibodies derived from lupus-prone mice. Six other clonally unrelated anti-DNA antibodies from the literature or from data banks expressed nearly identical VH genes (< or = 4 nucleotide differences) and eight others had nearly identical protein sequences (< or = 3 amino acid differences). Analysis of the germ line with oligonucleotide probes from the CDR regions suggests that all 10 autoantibodies are derived from a single member of the J558 gene family, which is present only in mice with the j haplotype for the J558 gene family. The amount of somatic mutation in these VH genes appears to be low, suggesting that some V, N, and D gene combinations can generate high affinity IgG anti-DNA auto-antibodies with little or no somatic mutation. Unusual reading frames, D-D fusions, and inversions were common in the IgG antibodies and may have been co-selected. Although the N and D regions of one IgM and all five IgG autoantibodies contained Arg residues, the presence of Arg residues was not correlated with binding to dsDNA or with pathogenicity. These results suggest that differences in the Ag-binding properties and the pathogenicity of these antibodies are determined by the CDR3 region and the L chain. PMID- 8409428 TI - Endogenous expression of delta on the surface of WEHI-231. Characterization of its expression and signaling properties. AB - WEHI-231 is a murine B cell lymphoma that has been used extensively as a model for the immature stage B cell and its functional response to Ag receptor cross linking as a model for immature B cell tolerance. This cell line expresses sIgM, CD5, and FcR gamma, but lacks the B cell-specific isoform of CD45 (B220). This study demonstrates for the first time that WEHI-231, in contrast to classically defined immature B cells, expresses delta on its surface. Analysis of delta on WEHI-231 revealed structural differences with respect to that on BAL-17 or primary splenic B cells. Although the m.w. of delta on the latter two B cell populations was similar, delta on WEHI-231 manifested a marked increase in its apparent m.w. deduced by SDS-PAGE. This difference was found to be due primarily to differential N-linked glycosylation. Signal transduction through the endogenous sIgD on WEHI-231 was investigated. In contrast to cross-linking of sIgM, cross-linking of the endogenous surface IgD on WEHI-231 was unable to generate a negative growth response in these cells. This inability may be due to uncoupling from normal surface Ig signaling pathways. The signaling properties of the endogenous sIgD on WEHI-231 differ from that on primary B cells and other sIgD-expressing cell lines. Whereas sIgD on splenic B lymphocytes or the mature B cell line BAL-17 is coupled to inositol phospholipid hydrolysis and calcium mobilization, cross-linking of sIgD on WEHI-231 failed to elicit these events, although induced changes in tyrosine phosphorylation were observed. Thus, endogenous expression of surface IgD on WEHI-231 is inconsistent with its representing the classically defined immature stage B cell. The structural and signaling differences associated with delta on these cells suggest the potential for developmentally regulated delta function and model for study of sIgD signal transduction. PMID- 8409429 TI - Antioxidant treatment of thymic organ cultures decreases NF-kappa B and TCF1(alpha) transcription factor activities and inhibits alpha beta T cell development. AB - Using electrophoretic mobility shift assays (EMSA), we have recently shown that nuclear extracts of 14-day mouse fetal thymocytes contain abundant NF-kappa B transcription factor activity. To determine the functional role of NF-kappa B in early thymocyte development, we have exposed fetal thymus organ cultures to inhibitors of NF-kappa B activation, namely the antioxidants N-acetyl-L-cysteine and butylated hydroxyanisole. Both compounds caused a dose-dependent arrest of thymocyte differentiation toward alpha beta, but not gamma delta, T cells. This was associated with a profound decrease in nuclear content of NF-kappa B and TCF1(alpha) transcription factor activity, as determined by EMSA. In contrast, NF Y was affected less strongly, and cyclic AMP-response-element-binding protein levels remained essentially unchanged by antioxidants. To test the idea that alpha beta T cell development is correlated with NF-kappa B and TCF1(alpha) activity, we conducted additional experiments in a submersion culture system in which the generation of alpha beta T cells can be manipulated. Standard submersion culture supports gamma delta but alpha beta T cell development. Under these conditions, EMSA showed that transcription factor activities were similar to those seen in the presence of antioxidants. Importantly, when the generation of alpha beta T cells in submersion culture was restored by elevating oxygen concentrations, there was a dramatic increase in TCF1(alpha) activity, and both NF-kappa B and NF-Y returned to control levels. Taken together, these results strongly suggest that NF-kappa B and TCF1(alpha), presumably in concert with other transcription factors, play an important role in the development of alpha beta T cells. PMID- 8409430 TI - Structural analysis of the CD3 zeta/eta locus of the rat. Expression of zeta but not eta transcripts by rat T cells. AB - We analyzed the structure and pattern of expression of rat TCR zeta- and eta chains to investigate if these components function in activation and development of rat T cells. The rat zeta cDNA contained the complete open reading frame coding for a polypeptide of 164 amino acids and the 5' and 3' noncoding sequences. Comparison of the amino acid sequence to those of mouse and human counterparts revealed a high degree of similarity, more than 85% homology among all three species except for the signal peptide, which was especially high in the cytoplasmic domain including the nucleotide binding site and the possible tyrosine phosphorylation sites. Furthermore, we determined the nucleotide sequences of a rat genomic eta-like sequence located in the 3' region of the rat zeta-gene. Although it showed a high level of nucleotide similarity to mouse and human counterparts, 90.4 and 78.9%, respectively, the deduced polypeptide was very short (only 28 residues) and markedly divergent from the mouse and human eta specific polypeptides due to frameshift mutations. Transcription of rat zeta was shown to be highly restricted to T cells; abundantly in thymocytes and scarcely in peripheral T cells. Surprisingly, the rat eta transcript could not be detected in any rat tissues so far tested by Northern blot analysis and even by the sensitive reverse transcription-polymerase chain reaction method, whereas it was readily detected in mouse thymus. These findings suggest that the zeta-chain has conserved roles in the TCR assembly and TCR-mediated signaling. However, the eta chain seems not to be indispensable because of its structural diversity among these three species characterized to date and the apparent lack of eta expression in the rat. PMID- 8409431 TI - Switch recombination breakpoints occur at nonrandom positions in the S gamma tandem repeat. AB - Ig switch (S) recombination is clearly focused on S regions. It is possible that S-specific DNA binding proteins facilitate the alignment of these regions before recombination. The S gamma 3-specific DNA binding proteins, SNAP and SNIP/NF kappa B, interact with two discrete regions of the S gamma 3 tandem repeat, the A and B sites. Recombination breakpoints in the S gamma 3 region were found to significantly correlate with the binding sites of the S gamma 3 binding proteins. We now report the conservation of the SNIP and SNAP binding sites in S gamma 2b and S gamma 1 DNA. SNIP/NF-kappa B interacts with its cognate sites in S gamma 2b and S gamma 1 DNA as determined by mobility shift assays, competition binding studies, and supershift analysis using an antiserum specific for the p50 component. SNAP binds specifically to S gamma 2b and S gamma 1 as measured by mobility shift assays and competition binding studies. SNAP is composed of two closely traveling mobilities that do not separate on partial purification. SNIP and SNAP are expressed in nuclear extracts derived from murine splenic B cell cultures stimulated with mitogen or mitogen and IL-4. No new DNA binding proteins specific for S gamma 1 tandem repeats are detectable in nuclear extracts from B cells stimulated with mitogen and IL-4. The sites at which recombination occurs in the S gamma 2b and S gamma 1 regions have been analyzed statistically and found to correlate with the SNIP/NF-kappa B and SNAP binding sites. Distinctions have been found regarding the use of DNA substrates within the tandem repeat between primary (mu-->gamma) and successive (gamma-->x) S recombination. PMID- 8409432 TI - Self-peptides from four HLA-DR alleles share hydrophobic anchor residues near the NH2-terminal including proline as a stop signal for trimming. AB - Naturally processed MHC class II-associated peptides proved to be heterogeneous in size, varying from 13 to 25 amino acids. Truncation variants suggested sequence motifs that afford the amino termini to be shifted for obtaining an alignment: a 9- to 11-residue core region that is bordered by primary anchor residues is surrounded by extra sequences of variable lengths and hitherto unknown functions. Herein we present bulk sequencing analyses of self-peptides from four HLA-DR alleles and HLA-DQw7 clearly showing that the length of most of the NH2-terminal preanchor sequence is limited to 1 to 3 residues. Most strikingly, proline is the dominant residue reappearing at positions 2 and 3 in any allele. Proline revealed to function as a stop signal for NH2-terminal trimming as well as a secondary anchor: crude cytosolic and endosomal peptide fractions could be processed by aminopeptidases in vitro, whereupon DR1 binding peptides with increased affinity were generated. In addition, aminopeptidase treatment of DR1: self-peptide complexes implied that proline together with sterical constraints of the MHC molecule do protect the peptides' NH2-termini from further processing, whereas their COOH-termini were accessible to cathepsin B processing. Finally, bulk sequencing profiles contained signals from further putative anchor residues clustering in the NH2-terminal region:tyrosine, phenylalanine, leucine, isoleucine, and valine are enriched at positions 2 to 4 in DR1, DR5, and DR6, however, at positions 4 to 6 in DR3. Isotype-specificity is demonstrated by DQw7 displaying glutamine and asparagine at position 2. Obviously, the degenerate occurrence of aromatic or aliphatic side chains close to the NH2-terminal guarantees for essential interactions with a hydrophobic pocket of the investigated DR molecules. Most probably, this pocket is located in the nonpolymorphic DR alpha-chain rationalizing previous findings of promiscuous peptide binding to different DR alleles. PMID- 8409433 TI - Purification and characterization of chimeric human IgA1 and IgA2 expressed in COS and Chinese hamster ovary cells. AB - Ag-specific chimeric human IgA molecules, of the two human subclasses, IgA1 and IgA2, have been expressed in two mammalian cell systems. Analysis of the secreted IgA molecules, purified in milligram quantities from stable Chinese hamster ovary transfectants by Ag affinity chromatography, has allowed a direct comparison of the biologic properties of the two subclasses. HPLC gel filtration analysis revealed that in both subclasses, the IgA molecules associate predominantly into dimers. The monomer units are presumed to interact noncovalently, inasmuch as no dimers are evident when the antibodies are subjected to SDS-PAGE. The recombinant antibodies are glycosylated, inasmuch as a lectin blotting procedure revealed that the H chains of both subclasses are recognized by Con A. When subjected to digestion by preparations of IgA1-specific proteases secreted by two pathogenic streptococcal strains, Streptococcus sanguis and Streptococcus oralis, the recombinant IgA molecules behave just as their natural equivalents. Thus, only the chimeric IgA1 molecule is cleaved, with the IgA2 remaining intact. In terms of interaction with natural effector molecules, both recombinant IgA isotypes were shown to interact with Fc alpha receptors on calcitriol-stimulated HL-60 cells with similar affinity, but neither antibody was found to interact with human C1q. The expression system described readily permits manipulation of the human IgA genes, which should lead to a fuller molecular understanding of how this important antibody mediates its function. PMID- 8409434 TI - Characterization of a novel high affinity human IL-7 receptor. Expression on T cells and association with IL-7 driven proliferation. AB - Although both unstimulated and activated human T cells express high affinity IL 7R, only activated T cells can proliferate to IL-7. This responsiveness may occur as a direct result of changes in the structure of the IL-7R during T-cell activation. We have previously demonstrated such changes by affinity cross linking studies, and have shown that unstimulated human T cells express a single IL-7R of 90 kDa, whereas activated T cells express an additional 76-kDa IL-7 binding protein. In this study the origin and function of the p90 and p76 molecules have been investigated. To determine the role of each of these receptors in IL-7 driven proliferation, IL-7R expression and proliferative capacity were monitored during mitogenic stimulation. These analyses showed that the ability of PBMC to proliferate to IL-7 correlated with expression of the p76 IL-7R, and not with expression of the p90 IL-7R. IL-7-driven proliferation is mediated via high affinity IL-7R, and accordingly, Scatchard analysis revealed that, like the p90 IL-7R, the p76 IL-7R bound IL-7 with dual (high; Kd 38 pM and low; Kd 360 pM) affinity. Deglycosylation studies showed that the p90 and p76 IL 7R are not simply differently glycosylated isoforms of a single receptor. In agreement, mAb to the previously cloned IL-7R were found to stain unstimulated T cells that express only the p90 IL-7R but not T-cell clones that express predominantly the p76 IL-7R. These antibodies also immunoprecipitated the cloned IL-7R as a 90-kDa species from both 125-I-surface-labeled resting and activated T cells, but were unable to precipitate the 76-kDa IL-7R. In addition, PCR analysis of p76-expressing cells could not detect splicing of the extracellular domain of the cloned IL-7R, thereby excluding the possibility that the p76 IL-7R is a previously undescribed splice variant of the cloned IL-7R. These data demonstrate that the p90 IL-7R is the T-cell homologue of the cloned IL-7R, and imply that the p90 and p76 IL-7R have different extracellular domains. Taken together these data suggest that the 76-kDa receptor is a novel high affinity IL-7R that may be necessary for IL-7 driven proliferation in human T cells. PMID- 8409435 TI - Biochemical identity of the mouse Ly-19.2 and Ly-32.2 alloantigens with the B cell differentiation antigen Lyb-2/CD72. AB - Lyb-2/CD72 is a 45-kDa mouse B cell surface protein that binds CD5 (Ly-1) and has been shown to induce B cell proliferation upon mAb binding. The serologically defined Ly-19.2 and Ly-32.2 lymphocyte alloantigens have mouse strain distribution patterns similar to that of the Lyb-2/CD72 alleles and map to the same region on chromosome 4 as Lyb-2/CD72. Our recent isolation of the Lyb-2a, 2b, and -2c cDNA has enabled us in this report to examine the relationship between Ly-19, Ly-32, and Lyb-2/CD72. A rat T cell line transfected with a mouse Lyb-2a cDNA is recognized by Ly-19.2-specific mAb, whereas transfectants expressing the Lyb-2b cDNA are recognized by both Ly-19.2 and Ly-32.2-specific mAb. Cell surface iodination immunoprecipitation analysis from Lyb-2a cDNA transfectants using Lyb-2a- and Ly-19.2-specific mAb as well as from Lyb-2b cDNA transfectants using Lyb-2b-, Ly-19.2-, and Ly-32.2-specific Ab, produced immunoprecipitates containing comigrating 45-kDa polypeptides. Preclearing studies with these transfectants indicate that the immunoprecipitated proteins represent the same polypeptide chain. These results demonstrate that the mouse Ly 19.2 and Ly-32.2 alloantigens are in fact the B cell differentiation Ag Lyb 2/CD72. PMID- 8409436 TI - Protective immunity in baboons vaccinated with a recombinant antigen or radiation attenuated cercariae of Schistosoma mansoni is antibody-dependent. AB - Mice vaccinated with radiation-attenuated cercariae of Schistosoma mansoni exhibit high levels of resistance to challenge infection. We have previously shown that sera from these mice recognize polypeptides that are expressed on the surface of newly transformed schistosomula. We have cloned and sequenced a cDNA that encodes a 62-kDa portion of one of these polypeptides. Vaccination of mice with this 62-kDa polypeptide (designated rlrV-5) elicits high antibody titers and significant resistance to challenge infection. We report here the results of a vaccination trial in baboons with the rlrV-5 or radiation-attenuated cercariae. rlrV-5 was presented either in the form of protein micelles or complexed with the outer membrane protein of meningococcus to form proteosomes. The level of protection achieved in these groups ranged from 0 to 54%, with a mean of 27.7%. In baboons exposed to radiation-attenuated cercariae the level of protection was very high, with a mean of 84%. The resistance observed after vaccination with rlrV-5 or radiation-attenuated cercariae was reflected in the overall histopathology. Vaccination of baboons with rlrV-5 or radiation-attenuated cercariae elicited an antibody response against epitopes exposed on the surface of newly transformed schistosomula. In the case of baboons vaccinated with radiation-attenuated cercariae, this response was not limited to epitopes encompassed by rlrV-5. Analysis of individual baboon sera by ELISA demonstrated that there was a direct correlation between the anti-rlrV-5 titer and resistance to challenge worm burden, suggesting that the immunoprotective mechanism is antibody-dependent. PMID- 8409437 TI - Expression of a protective intestinal immune response can be inhibited at three distinct sites by treatment with anti-alpha 4 integrin. AB - The alpha 4 integrins mediate lymphocyte adhesion to Peyer's patch high endothelial vessels and homing to Peyer's patch, as well as to mesenteric lymph nodes. In rats, immunity to infection with the nematode Trichinella spiralis is known to be mediated by CD4+ OX22- (CD45RC-) cells that home to the intestine in large numbers. These experiments were conducted to determine whether the alpha 4 integrins or LFA-1 were involved in the expression of intestinal immunity to T. spiralis. Injection of the anti-alpha 4 integrin, mAb TA-2, but not anti-LFA-1, mAb TA-3, impaired the expression of immunity. An effect of TA-2 was measured at three distinct sites along the activation pathway leading to the migration of protective CD4+ OX22- cells to the intestine. Injection of TA-2 on the same day as infection prevented normal rejection of the parasite and abrogated the characteristic appearance of blast cells in draining lymph 3 days after infection. A similar effect on the migration of blast cells at day 3 was seen when TA-2 was injected 1 day after infection, and injection 2 days after infection still reduced the number of protective cells entering TD lymph on day 3. The effect of TA-2 and TA-3 on homing of dividing cells to the gut was examined by injecting dividing cells i.v. at the same time as antibody. Under these conditions migration of dividing cells to the gut was reduced by 90 to 95% and their capacity to adoptively transfer worm rejection blocked. Furthermore, TA 2 treatment also inhibited protection when it was injected 12, 18, or 24 h after the transfer of protective cells, when these cells had already entered the gut, but not when TA-2 injection was delayed for 36 h. These results indicate the involvement of alpha 4 integrins at the following points in the generation, dissemination, and function of CD4+ OX22- effectors: 1) initial activation during the first 48 h of infection; 2) migration of protective cells to and extravasation in the gut; 3) a function after entry into gut tissues. The results suggest that entry of dividing cells into the gut is critical for the adoptive transfer of protection and that alpha 4 integrin has multiple roles in the manifestation of intestinal immunity. PMID- 8409438 TI - Spontaneous and IL-2-induced anti-leukemic and anti-host effects against tumor- and host-specific alloantigens. AB - The graft-vs-leukemia (GVL) effect and its connection to graft-vs-host disease (GVHD) were studied in F1 hybrid mice after parental bone marrow transplantation. (BALB/c x C57BL/6) F1 mice (F1) were sublethally and lethally irradiated and inoculated with 10(4) BCL1 cells, a murine leukemia of BALB/c (BALB) origin. Subsequently mice were transplanted with F1 (syngeneic), BALB (allogeneic to the host but syngeneic to the tumor), C57BL/6 (B6) (allogeneic to both host and tumor) spleen cells, or spleen cells mixed with syngeneic (F1) bone marrow cells, respectively. Fresh BALB (H-2d) and B6 (H-2b) spleen cells, which induce equally strong GVHD in the F1 recipient against H-2b and H-2d alloantigens, respectively, were assayed for their ability to induce GVL effects on BCL1 (H-2d) cells with and without in vivo administration of human rIL-2. In order to increase the GVL effects, spleen cells were incubated for 4 days with rIL-2 to generate IL-2 activated killer cells. The results of adoptive transfer experiments showed that only immunocompetent B6 cells led to consistent eradication of leukemia. Neither BALB nor syngeneic F1 cells induced curative GVL effects regardless of whether spleen cells were further stimulated with rIL-2 in vitro, in vivo, or both. Our data support the theory that GVL effects are caused by T cell-dependent immune responses of allogeneic T cells and not by MHC-nonrestricted NK or rIL-2 activated NK cells, independently of the GVHD process itself. Hence, recognition and killing of tumor cells is most probably a MHC-restricted T cell-dependent process. PMID- 8409439 TI - Profiles of cytokine production in relation with susceptibility to cerebral malaria. AB - Infection with Plasmodium berghei ANKA (PbA) leads, in susceptible strains of mice, to the development of cerebral malaria (CM), a lethal syndrome that reproduces some features of human CM. To study a possible relationship between genetic susceptibility to CM and the cytokine expression pattern, we quantitatively evaluated gene expression on RNA extracted from various organs of malaria-infected mice, using strains that are susceptible and resistant to CM. Northern blot analysis and semi-quantitative PCR showed that CM is associated with an increased TNF-alpha mRNA accumulation in the brain of mice developing the neurologic complications of CM. An increased IFN-gamma mRNA accumulation and a decreased expression of IL-4 and TGF-beta genes were also observed in mice susceptible to CM. In vitro restimulation studies using crude malarial Ag showed that lymphoid cell proliferation was higher in CM-susceptible than in CM resistant infected mice. Moreover, susceptible mice produced large amounts of IFN gamma, in a dose-dependent manner, in response to PbA Ag, whereas cells from resistant mice failed to produce significant amounts of this cytokine. Conversely, IL-2 and IL-4 production was significantly higher in infected CM resistant mouse cells. No difference was seen in the production of IL-3 and IL-5 between resistant and susceptible PbA-infected mice. Upon stimulation with various malarial Ag, comparable amounts of TNF-alpha were produced by macrophages of either strain of mice. Taken together, these findings indicate that susceptibility to CM resides at the level of T cells rather than macrophages. Furthermore, the cytokine production profile is consistent with a predominant Th1 like response in mice developing cerebral complications of malaria. PMID- 8409440 TI - Killing of Staphylococcus aureus by tumor necrosis factor-alpha-activated neutrophils. The role of serum opsonins, integrin receptors, respiratory burst, and degranulation. AB - We have examined the effects of TNF priming on the killing of Staphylococcus aureus by human neutrophils. In the absence of serum opsonins, neutrophils failed to kill S. aureus, and TNF priming did not induce the cells to become bactericidal. Normal human serum, containing complement activity, promoted the killing of the bacteria by neutrophils. Pretreatment of neutrophils for 30 min with TNF significantly enhanced their bactericidal activity. The effects of TNF on neutrophil bactericidal activity was dependent on serum concentration and the degree of enhancement induced increased up to a concentration of 1%. The kinetics of bacterial killing showed that TNF-only enhanced the initial rate of killing, over the first 30 min. Little killing of bacteria occurred in the presence of complement-inactivated serum, and TNF did not stimulate this killing. These results suggest that TNF enhances the neutrophil complement-dependent killing of S. aureus. TNF increased the expression of CR3 (CD11b/CD18) and CR4 (P150, 95; CD11c/CD18) adhesion receptors but not LFA-1 (CD11a/CD18); and mAb against the alpha-chain of either CR3 or CR4 but not LFA-1 prevented the enhancing effects of TNF on the neutrophil bactericidal activity. PMID- 8409441 TI - Inhibition of murine B cell proliferation and down-regulation of protein kinase C levels by a phosphorylcholine-containing filarial excretory-secretory product. AB - E-S 62, a major excretory-secretory product of the rodent filarial parasite, Acanthocheilonema viteae, inhibits the polyclonal activation (DNA synthesis) of mouse B cells by mitogenic anti-Ig antibodies. This effect appears to be due at least in part to the phosphorylcholine (PC) moiety of the molecule, because it can be mimicked by PC-BSA or PC-chloride. Activation of the B cells by LPS is not influenced by the presence of E-S 62/PC, suggesting that they may target some aspect of signaling via the Ag receptor. E-S 62/PC failed to inhibit surface Ig mediated generation of the second messenger, inositol triphosphate, indicating that their target may not be the early biochemical events associated with activation. Exposure to E-S 62/PC was found to lead to a reduction in the level of protein kinase C, an important downstream regulatory enzyme, in anti-Ig treated cells. This protein-kinase C down-regulation may be the biochemical mechanism underlying E-S 62/PC-mediated inhibition of surface Ig-activated B cells. PMID- 8409442 TI - Involvement of TCR-V beta 8.3+ cells in the cure of mice bearing a large MOPC-315 tumor by low dose melphalan. AB - We have previously shown that the curative efficacy of low dose melphalan (L phenylalanine mustard; L-PAM) for mice bearing a large s.c. MOPC-315 tumor requires the participation of CD8+ (but not CD4+) T cell-dependent antitumor immunity. Here we show that CD8+ T cells obtained from regressing tumors on day 4 or 5 after low dose L-PAM therapy of MOPC-315 tumor bearers (L-PAM TuB mice) display a preferential enhancement in the utilization of the TCR-V beta 8.3 gene segment as compared to CD8+ T cells from normal lymph nodes. Treatment of L-PAM TuB mice with mAb F23.1, which leads to the depletion of V beta 8.3+ cells, as well as V beta 8.1 and 8.2+ cells, led to a significant reduction in the ability of their tumor-infiltrating lymphocytes as well as their spleen cells to lyse MOPC-315 tumor cells in vitro in a short term assay. In addition, the mAb F23.1 treatment almost completely abrogated the lytic activity of the tumor infiltrating lymphocytes against another syngeneic, antigenically related plasmacytoma (the MOPC-104E). Moreover, the mAb F23.1 treatment significantly reduced the curative effectiveness of low dose L-PAM for mice bearing a large MOPC-315 tumor. In contrast, mAb KJ16 treatment, which leads to the depletion of V beta 8.1 and 8.2+ cells (but not V beta 8.3+ cells), did not reduce significantly the curative effectiveness of low dose L-PAM for such MOPC-315 tumor bearers. Thus, V beta 8.3+ T cells are important for the curative effectiveness of low dose L-PAM therapy for MOPC-315 tumor bearers, and it is conceivable that the V beta 8.3+ cells mediate their effect (at least in part) by contributing to the acquisition of CTL activity against plasmacytoma-shared Ag. PMID- 8409443 TI - An immune suppressive factor derived from esophageal squamous carcinoma induces apoptosis in normal and transformed cells of lymphoid lineage. AB - An immunosuppressive factor produced by an esophageal squamous carcinoma cell line mediates profound irreversible suppression of in vitro proliferative responses of lymphoid cells. Exposure of activated normal PBL to the immune suppressive factor (ISF) resulted in the induction of an irreversible anergic state with apoptosis evident in 20% of those cells. Flow cytometric cell cycle analysis of mitogenically stimulated normal lymphocytes exposed to the ISF revealed that despite exhibiting full activation status (IL-2 production, IL-2R, and transferrin receptor expression) PBL were arrested at the G1/S interphase of the cell cycle. Transformed lymphoid cell lines, NSO and JURKAT, displayed morphologies characteristic of apoptosis within 24 h of exposure to the ISF. Flow cytometric cell cycle analysis of the JURKAT cells incubated with ISF revealed that > 90% of these cells had undergone apoptosis within 24 h. Agarose gel electrophoresis of DNA extracted from the ISF-treated lymphoid cells resolved a DNA fragmentation pattern characteristic of apoptosis in both the NSO cells and to a lesser extent in the activated PBL exposed to the ISF but not in control cells. In JURKAT cells stimulated with anti-CD3 antibodies, Ca2+ mobilization was markedly enhanced in those cells exposed to ISF. Also, ISF independently induced a calcium flux in JURKAT cells. Induction of programmed cell death by ISF may account for the in vivo immune suppression local to the tumor site in squamous carcinoma of the esophagus. PMID- 8409444 TI - Modulation of murine cytokine responses to mycobacterial antigens by helminth induced T helper 2 cell responses. AB - BALB/c mice inoculated with live Brugia malayi microfilariae (mf), or immunized with a soluble filarial extract (B. malayi Ag (BmA)), develop a pronounced Th2 like response over time. In contrast, single or repeated immunizations with a soluble Mycobacterium tuberculosis Ag preparation (purified protein derivative, PPD) stimulates a Th1, but not Th2 response (IFN-gamma >> IL-4, IL-5). To determine if the Th1 response to PPD can be modulated by the ongoing helminth induced Th2 activity, mice were: 1) immunized simultaneously with BmA and PPD; 2) immunized first with BmA, then with PPD; or 3) inoculated with live mf and immunized with PPD at various times thereafter. Simultaneous immunization with both Ag had no effect on the Th response induced by PPD, i.e., it was strictly Th1. In contrast, establishment of a Th2 response by either inoculation of live mf or immunization with BmA before administration of PPD skewed the PPD-specific Th response such that IL-4 and IL-5 were produced in addition to IFN-gamma. IL-4 and IL-5 levels produced in response to PPD under these conditions were further elevated in vitro in the presence of neutralizing IFN-gamma. Finally, in vivo neutralization of IL-4 diminished induction of Th2 responses to PPD. These results demonstrate that ongoing Th2 responses to helminth Ag modulate the Th response to mycobacterial Ag by an IL-4 dependent mechanism. PMID- 8409445 TI - Inhibition of human CTL-mediated lysis by fibroblasts infected with herpes simplex virus. AB - Previously, we demonstrated that human anti-HSV CTL and allo-antigen-specific CTL were inhibited in lysing their normally sensitive target cells when they were exposed to human fibroblasts (FB) infected with herpes simplex virus (HSV). In this study, the mechanism of inhibition of CTL lytic function by FB infected with HSV-1 (HSV-FB) was studied. CTL exposed to HSV-FB early (2 h) in the infection cycle were inhibited by a mechanism that appears to be distinct from the inhibition of lytic function mediated by HSV-FB at late times (20 h) during the infection cycle. The inhibition of CTL-mediated lysis by FB infected with HSV-1 for 2 h required the expression of ICP4, an immediate-early protein of HSV-1, but not the production of infectious virus or virus-induced shut-off of host protein synthesis. In contrast, the expression of HSV-specific glycoproteins essential for viral infectivity (glycoproteins B, D, H, K, and L), and thus, infectious virus, was required for inhibition of CTL lytic function by FB infected with HSV 1 for 20 h. Further, CTL exposed to FB infected with HSV-1 for 20 h expressed HSV specific proteins indicating that they were infected with HSV-1. Cell-to-cell spread of HSV-1 appeared to be the major mode of transmission because 1) an insufficient level of HSV-1 was present in the supernatant of HSV-FB to inhibit CTL lytic function; and 2) paraformaldehyde-fixed HSV-FB did not inhibit CTL mediated lysis. The inhibition of CTL lytic function by HSV-FB may be an important mechanism of HSV-induced immunosuppression, permitting the virus to spread and persist in immunocompetent hosts after primary infection or reactivation of latent HSV. PMID- 8409446 TI - Transforming growth factor-beta expression and natural killer cell responses during virus infection of normal, nude, and SCID mice. AB - The experiments presented here assess the contribution of T cells to NK cell regulation and transforming growth factor-beta (TGF-beta) production. The kinetics of NK cell activation in the absence of T cells was evaluated by comparing responses in athymic nude and SCID mice to those in normal C3H/HeN, C57BL/6, and BALB/c mice during lymphocytic choriomeningitis virus (LCMV) infection. T cell-deficient mice exhibited high NK cell activity for longer periods of time post-infection. To determine whether the reduction in NK cell activation at later times post-infection of normal mice resulted from induction of negative regulatory elements, mice were treated with the chemical IFN and NK cell inducing agent polyinosinic-polycytidylic acid (poly I:C). Although poly I:C induced plasma IFN in both uninfected and LCMV-infected normal mice, NK cell activity was elevated in the uninfected mice only. In vivo expression of TGF-beta protein in normal and T cell-deficient mice was evaluated by immunohistochemical studies on splenic sections. TGF-beta proteins were not detected in sections from uninfected mice but were induced in all strains during infection. High levels of TGF-beta proteins were localized to discrete cells, and diffuse staining was observed along marginal zones. The ability of the splenic leukocytes to release TGF-beta into conditioned media was evaluated in the Mv 1 Lu biologic assay. Transient acid treatment to release active factor from the latent form and use of specific neutralizing antibodies demonstrated that leukocytes from all strains of infected mice had released latent TGF-beta 1 into conditioned media. However, cells from infected nude or SCID mice produced substantially less active TGF-beta than those isolated from normal mice. Production of biologically active TGF-beta 1 was dependent upon the presence of T cells in normal mice, as in vivo depletion of CD4+ and CD8+ T cells by antibody treatment resulted in augmented NK cell activity and loss of active factor production. The lack of TGF-beta production was accompanied by increased NK cell numbers; at late times post-infection, T cell-deficient C57BL/6-nude mice had almost twice as many NK1.1+CD3- cells per spleen compared with T cell-containing C57BL/6 littermate control mice. Treatment with active TGF-beta 1 reduced the numbers of NK cells per spleen in infected nude mice to the levels found in infected littermate controls.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8409447 TI - Resistance of mice to experimental leishmaniasis is associated with more rapid appearance of mature macrophages in vitro and in vivo. AB - Resistance to murine leishmaniasis has been related to the propagation of specific Th cell subsets (Th1 and Th2). This study shows that there are differences between resistant and susceptible mice in the initial myelomonocytic infiltrate, which precede the specific T cell response. After subcutaneous injection of 2 x 10(7) Leishmania major into footpads of resistant C57Bl/6 and susceptible BALB/c mice we performed immunohistochemical studies on the infiltrate. Two days after infection the percentage of more mature, F4/80 positive macrophages in the lesion increased faster in C57Bl/6 mice (63%) than in BALB/c mice (29%). The same strain-specific differences were observed after infection of corresponding strains of athymic mice (57.2% in C57Bl/6 nu/nu; 33.6% in BALB/c nu/nu), thus excluding a T cell-controlled phenomenon. After 1 wk the infiltrate in susceptible mice began to reveal significantly more cells containing MRP14, which is expressed by granulocytes and less mature monocytes but not by mature macrophages. No corresponding differences were found between athymic strains, suggesting that at this point organization of the infiltrate falls under control of protective T cells. In bone marrow cultures of BALB/c and C57Bl/6 mice, the percentage of F4/80-positive macrophages was also increasing faster in C57Bl/6 mice than in BALB/c mice. Increased expression of the F4/80 Ag was associated with higher leishmanicidal activity of C57Bl/6 macrophages. MRP14 positive bone marrow cells on the other hand were rarely infected by parasites. We suggest 1) that the earlier appearance of leishmanicidal macrophages in lesions of C57Bl/6 mice could influence propagation of either Th1 or Th2 cells by reduction of parasite load or by differential secretion of decisive cytokines and 2) that the diffuse accumulation of granulocytes and inflammatory monocytes in susceptible mice facilitates spread of disease. PMID- 8409448 TI - IL-4 inhibits growth factor-stimulated rheumatoid synoviocyte proliferation by blocking the early phases of the cell cycle. AB - A major feature of rheumatoid arthritis is an uncontrolled proliferation of synoviocytes. This is consistent with the active production of factors such as platelet-derived growth factor (PDGF) and IL-1 by the synovitis, which act in vivo as well as in vitro as potent synoviocyte growth factors. We have previously shown that IL-4 is able to inhibit growth factor production in an ex vivo model of synovitis. Herein, we show that IL-4 strongly inhibited PDGF and IL-1 beta stimulated rheumatoid arthritis synoviocyte proliferation in a dose-dependent manner and through its 130 kDa receptor. This antiproliferative effect of IL-4 directly correlated with a blockade of the synoviocyte cell cycle at the G0 + G1 phases. We also observed that IL-4 induced striking morphologic changes in IL-1 beta or PDGF-stimulated synoviocytes, including increased volume and granulosity. These changes led to major perturbations of the cell monolayer, associated with a marked decrease of synoviocyte viability. Taken together, these data indicate that IL-4 inhibits growth factor-induced proliferation of synoviocytes by interfering with the cell cycle, and by decreasing cell survival. PMID- 8409449 TI - Phospholipase A2 activity in human neutrophils. Stimulation by lipopolysaccharide and possible involvement in priming for an enhanced respiratory burst. AB - Exposure to LPS, platelet-activating factor, certain cytokines, and other agents can prime human neutrophils for an increased release of superoxide anion (O2-) in response to stimuli. Previous work with LPS has suggested that priming may involve alterations in signal transduction pathways related to the release of O2 . Products derived from the enzymatic activity of phospholipase A2 (PLA2) on membrane phospholipids reportedly activate certain of these signaling events. Thus, PLA2 could play a regulatory role in the release of O2- by neutrophils. We examined this possibility by studying the effect of LPS on neutrophil PLA2 activity. Exposure to LPS triggered a fivefold increase in activity of an endogenous PLA2; control cells incubated under identical conditions without LPS showed no increase. Neutrophil-associated PLA2 activity increased 5 to 10 min after the addition of LPS to the cells and preceded the emergence of the primed state. Quinacrine and p-bromophenacylbromide, inhibitors of PLA2, blocked LPS priming but not the baseline O2- release from unprimed cells. The LPS-provoked increase in PLA2 activity resulted in release of oleic acid (38 +/- 4% above baseline) but not arachidonic, linoleic, or palmitic acid. In contrast, ionomycin resulted in significant release of all four fatty acids. The addition of exogenous PLA2 to neutrophils primed them. Priming was rapid and was 80 +/- 5% complete within 3 min. Thus, LPS and perhaps other agents may mediate their effects on O2- release at least in part by triggering PLA2 activity. PLA2 activation may play a role in regulating production and release of O2- by the human neutrophil. PMID- 8409450 TI - Study of natural lipocortin I. A potent mediator for macrophage-mediated immunosuppression in tumor-bearing mice. AB - We previously reported a possible role of lipocortin I secreted from Mac-1+, -2+ macrophages in immunosuppression in tumor-bearing mice. In this study, we purified natural lipocortin I from the spleens of tumor-bearing mice and compared its immunosuppressive activity with recombinant lipocortin I produced by Escherichia coli. The culture supernatants of splenic macrophages from tumor bearing mice suppressed the mitogenic responses in splenic lymphocytes. The culture supernatants contained higher levels of lipocortin I, but not PGE2, which was considered as a major immunosuppressive factor. Anti-lipocortin I antiserum neutralized these inhibitory activities. Natural lipocortin I purified from the spleens of tumor-bearing mice showed a potent suppressive activity, which was > or = 200 times higher than that of recombinant mouse lipocortin I at protein level. Alkaline phosphatase treatment of natural lipocortin I decreased immunosuppressive activity to the level of recombinant lipocortin I. N glycosidase F treatment failed to decrease the immunosuppressive activities. These results suggest that the protein modification of lipocortin I, possibly phosphorylation but not glycosylation, is critical for the potent immunosuppressive activity in tumor-bearing mice. PMID- 8409451 TI - Regulation of mouse bone marrow macrophage mannose receptor expression and activation by prostaglandin E and IFN-gamma. AB - The macrophage mannose receptor mediates the clearance of microorganisms and glycoproteins containing terminal mannose oligosaccharides. Cell surface expression of this receptor progresses with macrophage differentiation, and thus may be critical to the scavenger function of tissue and circulating macrophages. Bone marrow macrophages, which were used in this study, differentiate in culture and express functional mannose receptors. The cytokine IFN-gamma triggered activation of these macrophages and down-regulated cell surface expression of the mannose receptor after 48 h. Macrophage activation, as assessed by the generation of superoxide radicals, was inversely correlated with mannose receptor expression. The number of surface receptors was diminished by exposure to IFN gamma, whereas the binding affinity of the mannose receptor remained unchanged. Treatment with IFN-gamma reduced receptor biosynthesis yet did not alter receptor degradation. Mannose receptor biosynthesis is up-regulated by PG of the E series, and these anti-inflammatory agents reversed the effects of IFN-gamma on receptor expression. Down-regulation of the mannose receptor by IFN-gamma was fully reversible by PGE, indicating that receptor levels are dependent on the functional state of the cell rather than being linked to terminal cell differentiation. The regulation of the receptor by cytokines and anti inflammatory reagents suggests that the mannose receptor plays a critical role in macrophage scavenger functions and potentially in modulating inflammatory reactions. PMID- 8409452 TI - GM-CSF regulates human eosinophil responses to F-Met peptide and platelet activating factor. AB - Evidence is accumulating that suggests that granulocyte macrophage CSF (GM-CSF) may have inflammatory actions in addition to its effects as a hematopoietic growth factor. In these studies, the effects of culture of purified human eosinophils with GM-CSF on cellular responses, including intracellular calcium responses, the expression of adherence molecules, and adhesion, were examined. In freshly isolated eosinophils expression of the adhesion molecule CD11b was only weakly increased by a 10-min incubation with platelet-activating factor (PAF) (118 +/- 3, 121 +/- 4, 117 +/- 2, n = 8) and FMLP (100 +/- 1, 106 +/- 6, and 105 +/- 4% of control for 10(-8), 10(-7), and 10(-6) M, respectively, n = 4). Lyso PAF never increased CD11b expression. Exposure of eosinophils to GM-CSF (0.5 ng/ml for 2 days) induced an increase in CD11b responses to PAF (132 +/- 3, 144 +/- 4, and 140 +/- 7) and FMLP (123 +/- 8, 145 +/- 10, and 137 +/- 4% of control for 10(-8), 10(-7), and 10(-6) M concentrations, respectively, n = 4). In 3-day culture studies, responses were greatest on days 2 and 3. The half-maximal GM-CSF concentration for the increase in the response to FMLP (10(-7) M, day 2) was approximately 100 pg/ml. GM-CSF (0.5 ng/ml) induced an increase in basal CD11b expression by day 1, which declined toward the initial baseline on days 2 and 3. Dexamethasone (10(-7) M) hastened the decline in baseline CD11b expression but did not inhibit the increase in the response to either PAF or FMLP. After 3 days of culture with GM-CSF (0.08 ng/ml), FMLP (10(-6) M)-induced elevations of intracellular calcium were increased (277 +/- 27% of day 0 control, n = 4, p < 0.01), whereas the response induced by PAF was not (70 +/- 36% of day 0 control, p = NS). A 30-min exposure to GM-CSF caused a potentiation of both PAF- and FMLP induced adherence of eosinophils to gelatin-coated plastic. Longer incubation with GM-CSF (3 days) potentiated only FMLP-induced adherence. We conclude that expression of adherence molecules, transmembrane signaling, and adherence responses of eosinophils are enhanced by exposure to GM-CSF. These effects of GM CSF could potentiate the adherence to endothelium and subsequent migration of eosinophils to inflammatory sites in vivo. PMID- 8409453 TI - Immortalization with herpesvirus saimiri modulates the cytokine secretion profile of established Th1 and Th2 human T cell clones. AB - T blasts of six established human CD4+ T cell clones with defined Ag specificity and cytokine secretion profile (3 Th1 and 3 Th2) were immortalized with Herpesvirus saimiri (HVS) and compared with their uninfected counterparts for their ability to proliferate, produce cytokines, and express cytolytic activity. HVS-transformed Th1 and Th2 clones neither substantially changed their original surface markers nor lose their ability to proliferate in response to their specific Ag but did acquire the ability to proliferate in response to contact signals delivered by SRBC or autologous APC alone. In addition, transformation by HVS substantially enhanced the lectin-dependent cytolytic activity of Th1 clones and enabled noncytolytic Th2 clones to exert cytolytic activity. HVS-transformed Th1 clones but not their uninfected counterparts spontaneously transcribed and secreted Th1-type cytokines (IL-2, IFN-gamma, and TNF-beta) and such a production was further enhanced by stimulation with either SRBC or PMA plus anti-CD3 mAb. HVS transformed but not uninfected Th2 clones constitutively expressed both IL-4 and IL-2 mRNA and secreted IFN-gamma. Stimulation with PMA plus anti-CD3 mAb induced uninfected Th2 clones to secrete high amounts of IL-4 and IL-5 but not Th1-type cytokines, whereas the same HVS-transformed Th2 showed minimal IL-4 and IL-5 secretion with concomitant high production of IL-2, IFN-gamma, and TNF-beta. Transformation by HVS also resulted in up-regulation of TNF-alpha and IL-3 production by both Th1 and Th2 clones. The ongoing proliferation of HVS transformed clones was partially inhibited by either anti-IL-2 or anti-IL-3 antibodies and virtually abolished by the combined addition of the two anticytokine antibodies, suggesting that both IL-2 and IL-3 can function as autocrine growth factors for HVS-transformed Th1 and Th2 clones. PMID- 8409454 TI - Elevated IFN-gamma and decreased IL-2 gene expression are associated with HIV infection. AB - Because cytokines have a central role in the regulation and function of the human immune system, expression of several key cytokine genes in HIV infection was compared by quantitative polymerase chain reaction studies in lymphocytes from HIV-seronegative and -seropositive subjects. Elevated levels of IFN-gamma mRNA and lowered IL-2 mRNA were found in the PBMC of eight seropositive men with CD4 T cells over 500/mm3 (mean, 647/mm3), whereas IL-4 and IL-10 mRNA were not changed significantly. PBMC obtained 2 yr later from four of these patients with stable disease status (unchanged CD4 T cell number) showed median mRNA levels that were nearer normal for IFN-gamma and for IL-2. Four other men whose CD4 levels fell more than 200/mm3 in the following 2 yr, however, showed increased IFN-gamma and lowered IL-2. Purified CD4 and CD8 T cells from 10 HIV-seropositive and 10 seronegative homosexual men were compared. Cytokine gene expression was found to be markedly different in CD4 and CD8 T cells from HIV-seropositive men. In CD8 T cells on a per-cell basis, the levels of cytokine mRNA were substantially lower than in CD4 T cells and were not markedly changed in HIV infection. In the CD4 T cells, on a per-cell basis, the mean mRNA levels of IFN-gamma, IL-10, and TNF alpha were increased substantially (p < 0.001) in HIV infection. IL-2 gene expression was not increased significantly. Thus, the low IL-2 mRNA expression seen in PBMC is primarily due to the reduced CD4 T cell numbers. Increased expression of IFN-gamma genes in CD4 T cells, however, indicates that these cells may be responsible for substantial amounts of circulating IFN-gamma that occur in HIV infection. The striking difference in the effect of HIV infection on the expression of IFN-gamma and IL-2 genes indicates that these cytokines are under separate control. IL-4 mRNA levels were not changed. IL-10 gene expression, however, was increased more in early HIV infection, with less of an increase later. Expression of all cytokines in CD4 T cells appeared to subside late in HIV infection. However, the balance of cytokine expression was altered in all stages of HIV infection. PMID- 8409455 TI - Induction of peri-insulitis but not diabetes in islet transplants expressing a single foreign antigen. A multi-stage model of disease. AB - We have created a new transgenic model in which the human Epstein-Barr virus receptor, CR2 (CD21), has been expressed in pancreatic beta-cells. Mice derived from three transgenic founders bred into H-2b (C57BL/6) or H-2k (CBA) genetic backgrounds did not become spontaneously diabetic. After transplantation of CR2 expressing islets under the kidney capsule of genetically matched recipients, a histologic picture of peri-insulitis was found. However, animals did not manifest cellular invasion of the islets, destruction of the islets, or diabetes for at least 230 days. Significant numbers of both T and B lymphocytes were found in the cell population surrounding the islets. A pronounced serologic response to CR2 was also present and appeared to precede the onset of peri-insulitis. Thus, in this model, we have separated the process of diabetes induction into at least two phases. One is associated with peri-islet infiltration and an antibody response. However, at least one second signal is likely necessary for the process of islet destruction to follow. PMID- 8409456 TI - CD4 mAb therapy modulates alloantibody production and intracardiac graft deposition in association with selective inhibition of Th1 lymphokines. AB - The accelerated (24 h) rejection of (LEWxBN)F1 cardiac allografts (Tx) in LEW rats sensitized with BN skin grafts, is abrogated with CD4 mAb (BWH-4) administration between skin (day -7) and heart (day 0) transplantation (Tx survival ca. 11 days, p < 0.0001). This study analyzed the effects of CD4 targeted therapy upon host IgG and IgM alloantibody (allo-Ab) within the serum by two-color flow cytometry, and within the Tx, by immunohistology. These data were correlated with mRNA and protein production profiles of Th1 (IL-2, IFN-gamma) vs Th2 (IL-4) specific cytokines (polymerase chain reaction and/or immunohistology). Skin grafts elicited a strong systemic IgM allo-Ab response, which peaked at the time of cardiac Tx rejection at 24 h. It was associated with extensive deposits of IgM on Tx endothelium. Treatment with BWH-4 mAb diminished circulating IgM allo-Ab levels, and only low levels of IgM could be detected at the Tx site. Conversely, the low circulating IgG allo-Ab levels during rejection at 24 h in untreated recipients were accompanied by a strong labeling for intra-Tx IgG. BWH 4 mAb therapy did not prevent totally the switch of the IgM to IgG, but the IgG allo-Ab response was earlier, less intense and more transient than in untreated recipients. Accelerated rejection triggered sequential lymphokine mRNA expression in cardiac Tx, with the peak of transcription for IL-2 (6-12 h) preceding that for IL-4 (24 h). Interestingly, although CD4 targeted therapy virtually ablated the induction of IL-2 mRNA, it preserved transcription of the IL-4 gene. BWH-4 mAb therapy decreased otherwise abundant intra-Tx IL-2 and IFN-gamma, but allowed a vigorous elaboration of IL-4, confirming the translation of mRNA to the protein in vivo. Thus, CD4 mAb-mediated abrogation of accelerated cardiac Tx injury correlates with suppression of Th1 responses (depressed IL-2 and IFN-gamma production), but sparing of the Th2 function (enhanced IL-4 elaboration). Indeed, CD4 mAb-induced allo-Ab depression and immunosuppressive effects may reflect selective targeting of proinflammatory Th1-like cells and the multifaceted effects of IL-4 produced by unopposed Th2-like cells. PMID- 8409457 TI - Abnormal selection of antibody repertoires in retrovirus-induced murine AIDS. AB - The mature B cell repertoire in the course of murine AIDS (MAIDS) was investigated. The polymerase chain reaction (PCR) was used to amplify a large diversity of rearranged Ig H chain genes in normal or infected mice, 2 and 8 wk after virus inoculation. Libraries were constructed from the polymerase chain reaction products. By sequencing V-D-J clones in these libraries and analyzing the respective complementary determining region 3 (CDR3), we have shown at 8 wk the emergence of a population of B cells with significantly less N diversity, some sequences lacking any N addition, a typical feature of fetal repertoires known for degeneracy, and autoreactivities. This decreased N diversity was not present 2 wk after inoculation and could not be related to a defect in terminal deoxytransferase expression because the steady-state levels of terminal deoxytransferase mRNA were found normal in MAIDS bone marrow 8 wk after inoculation. FACS analyses revealed a decreased number of bone marrow B cells (B220+, sIgM+) in MAIDS already present at 2 wk, suggesting an alteration in the pathway of B cell differentiation and resulting in a decrease of peripheral B cells renewal. A relative enrichment of spleen cells in long lived B cells as a consequence of this blockade may participate in the abnormal antibody repertoire selection occurring in MAIDS. These data suggest in the MAIDS pathogeny the relationship between an abnormal repertoire selection and the pathologic process. PMID- 8409458 TI - Complete Freund's adjuvant induces an earlier and more severe arthritis in MRL lpr mice. AB - A study was performed on the effect of CFA on the spontaneous arthritis of MRL lpr mice. The development of swelling and erythema was monitored for 1 mo after injecting 13- to 14-wk-old mice intradermally with CFA, at which time the histopathology of the joints and serologic responses to extracellular matrix proteins were investigated. In a series of six experiments, 67 to 82% of mice showed early clinical evidence of arthritis in contrast to the low percentage observed in control animals. Similarly, the histopathologic analyses on the CFA injected mice indicated a significantly higher frequency of advanced histopathologic alterations, characterized by cartilage erosion and pannus formation. The serologic evaluation of the adjuvant-injected mice demonstrated a significant enhanced antibody production to type I and type II collagens, DNA, and the Mycobacterium tuberculosis-positive control. This reproducible adjuvant enhanced model of murine arthritis will be extremely useful in evaluating experimental therapeutic regimes as the arthritis is initiated earlier and exhibits an enhanced frequency and severity compared with the spontaneous arthritis seen in MRL-lpr mice. PMID- 8409459 TI - Inhibition of inflammatory liver injury by a monoclonal antibody against lymphocyte function-associated antigen-1. AB - When mice were given an i.v. injection of LPS 7 days after an i.v. injection of Propionibacterium acnes, liver injury and a rapid increase of serum alanine aminotransferase and asparagine acid aminotransferase occurred. The in vivo administration of mAb against LFA-1 on days 1, 2, and 3 after the i.v. injection of P. acnes resulted in a potent inhibition of all these dysfunctions. Using P. acnes and the LPS model, we found that anti-LFA-1 mAb protected the mice from P. acnes and LPS-induced lethal shock. During the course of P. acnes and LPS-induced liver injury, inflammatory cells infiltrated the liver and caused a massive hepatic cell necrosis. Flow cytometry revealed that the liver-infiltrating cells were mainly leukocytes expressing a higher level of LFA-1 antigen than that seen in the normal liver. These results suggested that the LFA-1 molecule on liver infiltrating leukocytes may play an important role in the induction of inflammatory liver injury. PMID- 8409460 TI - Monoclonal antibody based ELISAs for measurement of activins in biological fluids. AB - Two sensitive monoclonal antibody (MAb)-based enzyme-linked immunosorbent assays (ELISAs), one for activin A (homodimer of beta A subunits) and one for activin B (homodimer of beta B subunits) in plasma have been developed. The activin A ELISA had an effective range of 0.2-50 ng/ml while the activin B ELISA's range was 0.1 25 ng/ml in human serum. Both ELISAs were specific with < 0.01% cross-reactivity with related hormones and follistatin (an activin binding protein), however the presence of recombinant human follistatin caused a decrease in measured level of activin A and B spiked human samples. The assay was linear across the standard curve range with intra- and interassay coefficients of variation were less than 15%. The level of activins in female serum range from 0.3 to 10.4 ng/ml. In summary, we have developed a reliable, convenient and rapid MAb-based enzyme immunoassay for determination of activin A and B levels in human serum which are also applicable for buffer, mouse and monkey serum matrices. This assay will be useful for studying the regulation and role of activin A and B in health and disease. PMID- 8409461 TI - Comparison of immunomagnetic beads coated with protein A, protein G, or goat anti mouse immunoglobulins. Applications in enzyme immunoassays and immunomagnetic separations. AB - Immunomagnetic beads were prepared using either protein A (PA) or protein G (PG) coupled to magnetic beads for binding antibodies at their Fc region. The performance of these beads was compared with commercially available beads coated with goat anti-mouse (G alpha M) immunoglobulins. Both the PA- and PG-beads possessed a higher binding capacity than the G alpha M-beads for the monoclonal antibodies tested, although, PA bound weakly with some IgG1 antibodies. PA-beads were compared with G alpha M-beads in a magnetic enzyme immunoassay for the detection of mouse immunoglobulins as an alternative to a conventional capture ELISA. The magnetic enzyme immunoassay was characterized by a detection time of less than 60 min and a linear assay range from 5-10 to 500 ng/ml for G alpha M beads and 5-10 to 1000 ng/ml for PA-beads. The capture ELISA was linear from 10 to 250 ng/ml. For immunomagnetic separation of Salmonella with immunomagnetic beads, PA-beads were superior to both PG- and G alpha M-beads. For specific isolation of bacteria from heterogeneous suspensions by immunomagnetic separation, PA- and PG-beads are preferable since G alpha M-beads crossreact with bacteria possessing proteins with Fc-binding activity. PMID- 8409462 TI - A quantitative radiometric assay to measure mammalian cell binding to hyphae of Candida albicans. AB - A rapid and reproducible assay has been developed to measure the capacity of lymphocytes to bind to Candida albicans. Lymphocytes that bound to C. albicans were either the large granular lymphocyte cell line, YT, or interleukin-2 activated lymphocytes. Lymphocyte binding was assessed as the associated radioactivity of 51Cr-labeled lymphocytes with preformed hyphae. The assay was sensitive to detection of 0.6 lymphocytes/one hyphal form at one half maximal lymphocyte binding capacity. The assay correlated well with direct microscopic assessment of lymphocyte binding to C. albicans and provided quantitative radiometric data. Although the assay was developed for the assessment of lymphocyte adhesion to C. albicans, it can be used to measure binding of other mammalian cells (e.g., polymorphonuclear leukocytes) to this fungus. In addition, the assay may be used to identify molecules involved in the adhesion of lymphocytes and other mammalian cells to C. albicans. PMID- 8409463 TI - In vivo labelling of the spleen with a red-fluorescent cell dye. AB - A method for labelling mouse spleen cells in situ is described. Spleen vessels were clamped before intrasplenic injection of a red-fluorescent cell dye (PKH 26). The labelling rate was 11.8% of all cells and 13.9% of B lymphocytes 30 min after injection as determined by FACS. 3 days later, the proportions of labelled cells were reduced to 7.4% (P < 0.01) and to 10.7% (P < 0.05), respectively. Only 10% of cells detected by FACS could be detected by fluorescence microscopy. Labelled cells were not found in peripheral lymph nodes 30 min after spleen injection, but were present 3 days later (FACS: 2.8% of all cells and 5.4% of B lymphocytes, P < 0.05 each), indicating migration of stained cells. Neither adverse nor toxic effects of in situ staining were observed. Isolated stained B lymphocytes from spleens of naive mice and from lymph nodes after immunisation with insulin showed proper function when tested in an ELISA-spot assay. The labelling procedure was used to follow splenic B lymphocytes producing natural anti-insulin antibody. These cells were found to be recruited for the early immune response in lymph nodes immunised with insulin. PMID- 8409464 TI - Affinity cleavage of cell surface antibodies using the avidin-biotin system. AB - In the present study, we have demonstrated the feasibility of targeting a proteolytic enzyme, via the high-affinity avidin-biotin system, to act in a highly selective manner upon a cell surface-associated antibody. As an example of this approach, a cell-bound biotinylated monoclonal antibody could be removed efficiently by means of biotinylated proteinase K, bridged to streptavidin. Only low levels of cell death were observed using this procedure. The approach may prove useful for a variety of applications, including the recovery of antibody free positively selected cell populations. PMID- 8409465 TI - Human natural killer cells: a convenient purification procedure and the influence of cryopreservation on cytotoxic activity. AB - The recognition of natural killer cells as a lymphoid subpopulation with a distinct set of surface markers has led to the development of a variety of antibody-based purification methods. In this paper we describe a rapid, three step negative selection protocol for the purification of human natural killer (NK) cells from the mononuclear cell fraction, which is obtained by the centrifugation of peripheral blood on Ficoll-Paque. Subsequently, monocytes and B lymphocytes are removed by adherence to nylon wool and T lymphocytes by panning with anti-CD3. With this procedure, CD3-, CD16/56+ NK cells are purified about five-fold, from 12 +/- 3% in the starting population to a final purity of 61 +/- 11%. A further increase to > or = 70% is obtained, if an extra Ficoll centrifugation step is included. The recovery of NK cells (50%) is significantly higher than is usually achieved by previously described procedures. Furthermore, we show that activation of cytotoxicity, with concomitant changes in target specificity, occurs when frozen/thawed NK effector cells are kept in culture in order to regain their pre-freezing cytotoxicity levels. PMID- 8409466 TI - Selection of anticoagulants for lymphocyte immunophenotyping. Effect of specimen age on results. AB - In a multi-center study, whole blood specimens from 31 HIV-positive and 43 HIV negative donors were collected in three different anticoagulants and assayed for lymphocyte subsets fresh (within 6 h), and 1 and 2 days later. Each center prepared the specimens by their routine whole blood lysis procedure, labeling with a recommended panel of two-color monoclonal antibody combinations. 1 day (up to about 30 h) after blood collection, the results obtained from blood collected in EDTA (ethylenediamine tetra-acetate), ACD (acid citrate dextrose), and heparin were similar to fresh. Up to 48 h, only ACD and heparin, not EDTA, yielded results similar to fresh specimens. These results were similar for both HIV positive and -negative specimens. PMID- 8409467 TI - Constructing polycompetitor cDNAs for quantitative PCR. AB - Analysis of mRNA levels using reverse transcription coupled with the polymerase chain reaction provides a powerful tool for studying cytokine regulation in cellular immunology. We report a novel method for cloning competitor cDNAs that is rapid, efficient and inexpensive. By linking multiple competitor cDNAs in tandem, polycompetitor constructs can be created that allow the use of a single reagent for individual PCR assays. Assays can be performed on minute samples of cell culture or tissue and can be reliably quantitated after routine gel electrophoresis without the use of densitometry or labeled nucleotides. The utility of this technique lies in the ability to produce a relatively inexpensive customized reagent that is simple to use and that allows for sensitive determinations of gene expression in a rapid and convenient manner. This method should allow investigators in many areas of biology to easily quantitate a broad range of important regulatory molecules. PMID- 8409468 TI - Rapid agglutination testing in an ultrasonic standing wave. AB - The time taken to perform diagnostic agglutination tests can be significantly reduced by applying an ultrasonic standing wave field to a droplet of reactants held in a capillary tube. Avian erythrocytes, bacteria and latex particles from commercially available test kits were agglutinated in 15 s, 5 min, and 1 min respectively. These times compare favourably with the times of 30 min, 4 h, and 8 min required for agglutination by the methods prescribed for the respective kits. No loss in sensitivity or specificity was observed with the ultrasonic method. A multi-test procedure is also described whereby a series of five droplets loaded in a single capillary can be tested in less than 4 min by drawing the capillary along the axis of the ultrasonic field of a ring transducer. PMID- 8409469 TI - Particulate nitrocellulose as a solid phase for protein immobilization in immuno affinity chromatography. AB - The use of nitrocellulose paper as a solid phase matrix for protein immobilization and its application in immunoaffinity chromatography is described. Pieces of nitrocellulose paper were frozen in liquid nitrogen and ground to a powder (NCP) which was then fractionated according to particle size by repeated sedimentation/resuspension cycles in water. The flow properties of different NCP fractions were compared with those of Sephadex G-50. Protein binding capacity and binding dynamics were investigated in a model system with bovine serum albumin (BSA) in phosphate buffered saline. Applications of the material are illustrated by batch chromatography for the removal of anti-carrier protein antibodies from a hapten antiserum and by affinity purification of a hapten-specific antibody fraction using NaSCN elution from haptenated NCP. Furthermore, the applicability of the material in column chromatography is demonstrated by elution of a monospecific fraction of anti-fluorescein antibodies from a hapten/carrier mixed bed column by excess of soluble hapten. The results demonstrate that NCP chromatography appears to be a cheap and useful alternative to many other chromatographic media used for protein immobilization. PMID- 8409470 TI - A modified method for obtaining large amounts of high titer polyclonal ascites fluid. AB - When limited amounts of antigens are available, it is often difficult to obtain large quantities of polyclonal antisera. Although antisera can be raised in small rodents, yields are usually small. To circumvent these problems, we have designed a modified method for generating polyclonal ascites fluid (AF). Using the appropriate strain of mice and adjuvant, we generated high serum titers by injection of 1-100 micrograms of protein. Following i.p. injection of compatible sarcoma cells, 13-19 ml of high titer (1:1000-1:20,000), polyclonal ascites fluid were obtained from each mouse. Similar results were obtained using nine different antigens. PMID- 8409471 TI - An improved method for generating single-chain antibodies from hybridomas. AB - Cloning the correct VL kappa gene from hybridomas derived from MOPC-21 can be problematic because such cell lines variably express a transcript which is aberrantly rearranged at the VJ recombination site. Cellular levels of the aberrant transcript can exceed that of productive light chain RNA, so a large proportion of the VL gene-derived products obtained on PCR amplification of hybridoma cDNA may not encode a functional protein. We have developed a method in which antibody variable region genes are recovered from hybridoma cDNA using a unique set of V gene family-specific primers; the V region genes are then spliced by PCR, in the form 5'-VL-LINKER-VH-3' (where the linker encodes [GlyGlyGlyGlySer]3), and cloned into an expression vector under control of T7 RNA polymerase. Plasmid DNA is isolated from colonies, and the insert is expressed in an in vitro rabbit reticulocyte lysate-based coupled transcription/translation system, in a microtiter plate format. Since aberrantly rearranged VL kappa genes contain a translation termination codon at amino acid position 105, only constructs containing the correctly rearranged gene produce a protein of the predicted size. We demonstrate the method by producing the single-chain form of OKT9, a murine IgG1 which binds to the human transferrin receptor, and extend the results to show that the protein generated by the in vitro expression system retains the antigen binding properties of the parent antibody. Our method will be generally useful for screening single-chain antibodies for function prior to large scale production in vivo. PMID- 8409472 TI - Rapid fluorometric quantification of monocyte attachment in tissue culture wells. AB - A simple fluorometric assay that permits rapid quantification of attachment of monocytes or macrophages in tissue culture wells is described. Using 4,6 diamidino-2-phenylindole (DAPI) as a specific fluorochrome marker for DNA, we observed a dose-dependent increase with strong linear correlation in fluorescent emission over a broad range of DNA concentrations. Measurements of the DNA content of the human monocytic cell line THP-1 demonstrated a linear correlation between fluorescence intensity and cell number from 5 x 10(4) to 1 x 10(6) cells, with an estimated average DNA content of 7.5 pg DNA per cell. While untreated THP 1 cells were not detectably adherent, PMA induction for 24 h results in 57-76% adherence to plastic surface. This method was found to be useful for measuring the number of peripheral blood monocytes separated from lymphocytes by attachment. 16 subjects were sampled and the standard deviation of each individual did not exceed 10%. The number of attached cells was between 10-16% of the total mononuclear cells. Fluorescence measurement of DNA with DAPI permits rapid and accurate determination of cell numbers and appears useful in the quantification of adherent populations such as myelocytic cells and cell lines. PMID- 8409473 TI - Purification of human IgG4 subclass with allergen-specific blocking activity. AB - Blocking antibodies (bAb) induced by allergen immunotherapy are restricted to the IgG1 and IgG4 subclasses, with IgG1 predominating early and IgG4 coming later. Study of IgG4 bAb has been limited, in part, by the absence of a method to purify IgG4. We describe a rapid immunoaffinity chromatographic method for the purification of that subclass from whole serum. Starting serum (TR) contained 90 micrograms/ml Dactylis glomerata (orchard grass) pollen (DGP)-specific IgG4, measured by indirect ELISA. The blocking activity of TR was assayed in vitro on IgE-sensitized human basophils. Immunoadsorption on a strong-binding anti-IgG4 monoclonal antibody (mAb) removed about 90% of the total and allergen-specific IgG4 and nearly all of the blocking activity from TR. An IgG4-rich fraction was then obtained by absorption of several small volumes of TR on a weak-binding anti IgG4 mAb column at neutral pH followed by elution with glycine-HCl buffer. The pooled eluates contained 82% IgG4, amounting to a 65-fold purification of the serum IgG4; the yield was approximately 30%. Nearly all the DGP-specific antibody was in the IgG4 component of the eluate. The blocking activity of the eluate was approximately equal to that of TR. Immunoblot patterns with the eluate and with TR on SDS-PAGE of DGP were nearly identical. This method thus provides a fully active, relatively pure IgG4 blocking antibody. Moreover, the results reinforce the importance of using a well-chosen mAb when purifying proteins by immunoaffinity chromatography. PMID- 8409474 TI - Trichobezoar. PMID- 8409475 TI - The performance and attitude of interns--a critical appraisal. PMID- 8409476 TI - Recent ordinance of Maharashtra government. PMID- 8409477 TI - Giant cell tumour of the metacarpal bone. PMID- 8409478 TI - Pneumatosis cystoides intestinalis. PMID- 8409479 TI - Pregnancy and childbirth in uterus didelphys. PMID- 8409480 TI - Wilson's disease with neurologic manifestation. PMID- 8409481 TI - Amelia. PMID- 8409482 TI - Ameloblastoma of maxilla. PMID- 8409483 TI - Worm obstruction. PMID- 8409484 TI - Fibroma of the tonsil. PMID- 8409485 TI - Late presentation of diaphragmatic rupture due to blunt trauma. PMID- 8409486 TI - Handle life with care: prevent violence and negligence. PMID- 8409487 TI - Biochemistry--the molecular tongue of medicine. PMID- 8409488 TI - Quantitation of proteinuria by use of single random spot urine collection. AB - Measurements of protein to creatinine ratios in single voided urine samples were compared with 24 hour urinary protein excretion for quantification of proteinuria in 20 healthy control and 30 patients of various renal diseases. The range of proteinuria in study group was 1.46-6.0 g/day and renal function was stable in all the cases. An excellent correlation was found between the results obtained by this method and those from the standard 24 hours urine collection method in healthy controls (r = 0.99) and in patients with renal disease (r = 0.98). In our study urinary protein-creatinine ratio in control group was less than 0.2 in all cases and in study group was more than 3.0 except in one. The results of the present study indicate that the random urine protein-creatinine ratio is highly reliable test for quantification of proteinuria in children and has obvious advantage in term of time, cost and patients convenience. PMID- 8409489 TI - Experience in elective hepatic resection. AB - A study was conducted on 35 elective hepatic resections performed by one surgical team over a period of 5 years with 14% postoperative mortality. The indications for hepatic resection were primary hepatocellular carcinoma in 20 cases (57%) and metastatic tumours from colorectal cancer in 12 cases (34%). Underlying cirrhosis of liver was found co-existent in 35% of patients of hepatocellular carcinoma. The 3-year actuarial survival rate after resection for HCC and metastatic tumour was 30% and 42% respectively. PMID- 8409490 TI - Amphotericin B therapy in kala-azar. AB - Twenty-seven adult patients of kala-azar unresponsive to sodium stibogluconate (SSG) were given amphotericin B in the dose schedule of 0.50 mg/kg body weight on alternate days for a total of 21 to 31 infusions, depending upon clinical and parasitological response. A 100% cure rate was achieved at the end of the therapy and no relapse was encountered during 12 months follow-up. Only mild adverse reactions were noted on the day of infusion during therapy. Amphotericin B was found to be an acceptable alternative drug for treatment of such cases. PMID- 8409491 TI - Prevention and early detection of oral, breast and cervical cancers: a practical approach in Indian context. AB - Cancer has emerged as a major killer disease in India. One-third of the total cancer load is constituted by oral, breast and uterine cervix cancers. Considerable knowledge is available on control strategies on these 3 major cancers and this can be transformed into the community through the individual health care practitioners. Oral cancer is preventable and both the precancerous and invasive lesions can be picked up easily by examination of the oral cavity. The occurrence of breast cancer in the country is on the increase. The poor survival rate indicates the impact of the major proportion of advanced cancer. An effective strategy comprising breast self-examination, examination by a physician and a mammography test, is described. Cancer of the uterine cervix is the commonest malignancy among females in several regions in India. An effective and definitive method is available in the form of Pap smear for the control of cervical cancer. An organised community screening programme is not economically viable at present. However, routine per speculum examination should be undertaken and this can result in clinical downstaging of cancer. Women above the age of 40 years should undergo regular Pap smear screening. Broad guidelines are presented for an opportunistic screening. PMID- 8409492 TI - Bronchial asthma and its treatment. PMID- 8409493 TI - Lipomatous tumour of uterus. PMID- 8409494 TI - Lessons from recent outbreaks of Vibrio cholerae non-01 diarrhoea in the Indian subcontinent. PMID- 8409495 TI - Passive smoking. PMID- 8409496 TI - Various protocols of cell culture antirabies vaccines. PMID- 8409497 TI - Ear-buds need cautious use. PMID- 8409498 TI - 68th All-India medical conference. Varanasi 1993.["Presidential Address"]. PMID- 8409499 TI - Laser in ophthalmology. PMID- 8409500 TI - Biomedical engineering: relevant or not? PMID- 8409501 TI - Histopathological and immunological studies in a cohort of balanitis xerotica obliterans. AB - Balanitis xerotica obliterans consisting of 52 cases had been studied in the present series. This disease entity is an insidious sclerosing disease of unknown aetiology, affecting mainly the skin and mucous membrane of glans, prepuce and sometimes the fossa navicularis urethrae or even terminal urethra. Atrophic white patches on external genitalia and obstructive uropathy are two common presenting features. It has been classified as local form of lichen sclerosus et atrophicus. Poor local hygiene, circumcision and association of auto-immune diseases like vitiligo are responsible as suggested in this study. Routine tests detected associated cystitis in 8 cases, haematuria in 4 cases, non-reactive VDRL in all cases and impaired glucose tolerance in 5 cases. Immunoglobulin profile (39 cases) showed altered pattern, suggesting some chronic antigenic stimulation. Out of 33 cases where biopsy done, histologically proved typical balanitis xerotica obliterans was found in 19 cases. Malignant change was noted in one case only. Circumcision and dilatation offered temporary relief. PMID- 8409502 TI - Fertility and reproduction following inversion of uterus. AB - Puerperal inversion of the uterus is itself a rare occurrence. Records of fertility and reproduction following an episode of uterine inversion are even rarer. The reproductive outcome following correction of uterine inversion in 6 cases seen over a span of 35 years is being reported here. One case of acute inversion was managed by manual replacement and the remaining 5 of chronic inversion were corrected by Haultain's operation. The only patient managed by manual replacement conceived thrice. She aborted once, but delivered 2 healthy live babies subsequently by caesarean section done for uterine inertia each time. Out of the 5 patients treated by Haultain's technique, 3 conceived later. One did not come for follow-up after the 5th month. Each of the other 2 had full-term normal delivery of living baby under supervision. None of the cases had any complications. It is concluded that even after operative correction of inversion, uncomplicated delivery may be anticipated. Conservative surgical management is ideal even in apparently neglected and chronic cases, as most of these women were primipara or multipara with only one or 2 living children and had a desire of future child bearing. PMID- 8409503 TI - Role of arthrography in non-traumatic painful wrist. AB - The arthrographic examination was carried out in 25 patients with non-traumatic painful wrist of more than 6 weeks duration. Plain x-rays were non-contributory in making any diagnosis in 20 cases. Arthrography was performed even when plain x rays were normal and it revealed the soft tissue abnormalities and early phases of cartilage destruction not visible on routine roentgenograms. The extent of synovial involvement demonstrated on arthrography proved useful when synovectomy was planned. After operation pain and swelling disappeared and functions of the joint improved considerably. PMID- 8409504 TI - Malignant orbital tumours: observation in north Bengal. AB - Sixty-one cases of malignant orbital tumours were analysed retrospectively in relation to the incidence, age, sex, race, anatomical site of origin of the tumours and their histological types in North Bengal. Retinoblastoma is the commonest type (65%) followed by adenocarcinoma (10%), squamous cell and basal cell carcinoma (8.8% each). Other tumours were rarely encountered. PMID- 8409505 TI - Efficacy of ciprofloxacin and norfloxacin in multidrug resistant enteric fever in adults. AB - Twenty-five out of 55 patients of enteric fever were documented as multidrug resistant enteric fever cases. In the resistant cases the drug sensitivity of salmonella species in vitro were mainly augmentin, ceftriaxone, aminoglycosides, ciprofloxacin and norfloxacin. Sixteen patients were treated with ciprofloxacin and 9 were treated with norfloxacin for 14 days. Fever touched normal in 62.5% cases with ciprofloxacin and in 33.3% cases with norfloxacin by the 7th day. It became normal in 88% with ciprofloxacin and in 66% with norfloxacin by the 10th day and it became normal in 100% cases in each group by the 14th day. The newer 4 quinolones can be recommended in multidrug resistant enteric fever in adults. PMID- 8409506 TI - Regression of choriocarcinoma. PMID- 8409507 TI - Low back pain--risk factors and prevention. PMID- 8409508 TI - Finance Act 1993: new income-tax amendments for tax-payers. PMID- 8409509 TI - Autotransfusion in neurosurgical operations. PMID- 8409510 TI - [Spontaneous fracture of an ureteral endoprosthesis]. PMID- 8409511 TI - [Multicystic renal dysplasia with ectopic implantation of the ureter in boys. Apropos of 6 cases]. AB - The prevalence of multicystic dysplastic kidney has increased since the development of antenatal ultrasound diagnosis. Even though, the rarity of symptomatic forms and case reports of spontaneous regressions antenatally or after postnatal ultrasound follow-up have reduced indications of nephrectomy which is less and less appropriate. However, discovery of ectopic ureteric implantation in boys makes certain additional investigations as intravenous pyelography, cystography and ultrasound, essential before making any therapeutic decision. Nevertheless, some cases remain undescriptible. In these cases, operative opacification of any patent ureter can provide more precise information about ureteric implantation. Lastly, only a dissection and resection as complete as possible of any patent ureter and pelvic cystic dilatation can prevent subsequent complications. PMID- 8409512 TI - [Microalbuminuria before and one month after extracorporeal lithotripsy. First results]. AB - Extracorporeal shock wave lithotripsy represents extraordinary progress for the treatment of urinary calculi. The apparent aggressivity of this technique leads to investigating whether it may have an impact on the renal parenchyma, as was suggested by many works, especially with radionuclide explorations. Microalbumin assays before and about one month after lithotripsy show an iatrogenic pathological increase in levels in some cases, especially after long, intensive sessions of treatment, whether the lithotripter used was of the shock-wave or piezoelectric-wave type. When lithotripty was performed with a piezoelectric-wave device, the increase in microalbumin is related to the "cumulative" factor, which is in turn related to the power, frequency (per second) and length of shots. More extensive investigations and, most importantly, investigations over a longer period are necessary to confirm or invalidate the results of this first study, to establish the frequency of cases where this abnormality appears, whether it can last long, and how far it can be predictive of the parenchymal lesions and of their long-term impact. PMID- 8409513 TI - [Urethral calibration in menopausal women examined for vesico-sphincter disorders]. AB - In order to appreciate the consequences of aging on the urethra, two prospective studies assessed its caliber in postmenopausal women. The population included aged women who came to consultation for disorders of the bladder and sphinecters. The clinical checkup, assessing the patient's age, morphology, time elapsed since menopause, local consequences of hormonal deficiency and urinary functional signs, was completed by an urodynamic examination. In the first study (76 patients), the urethra was calibrated using two Peters dilating bougies ranging from Charriere 10 to Charriere 26. In the second study (50 patients), measurements were made with OTIS-type ball dilators ranging from Charriere 9 to Charriere 30. This exploration was made in 126 patients. Urethral caliber was greater than or equal to Charriere 24 in 78% of patients. Only 2.3% of women had urethral "stenosis" with a Charriere score under 18. Contrary to what we were expecting, the urethral caliber does not seem to decrease with age. This unexpected statement and the difficulties of statistical analysis we have encountered lead us to complement our study. PMID- 8409514 TI - [Priapism. Apropos of 10 cases]. AB - The authors report a retrospective study collecting ten cases of priapism over fifteen years. Clinical diagnosis is evident. Idiopathic priapism is most frequent of all the medical treatments available, no one seems to be efficient. Among surgical procedures draining the venous blood from the cavernous bodies, spongiocavernous shunt (Al-Ghorab) has been a simple, fast and effective method in the last five patients. Functional result depends on the age and especially on the precocity of the surgical treatment. Therefore priapism is an andrologic emergency. PMID- 8409515 TI - [Urology in women: innovations and developments in 1992]. PMID- 8409516 TI - Conformational polymorphisms of cRNA of T-cell-receptor genes as a clone-specific molecular marker for cutaneous lymphoma. AB - A novel molecular assay for the detection and characterization of monoclonal lymphoid populations in clinical specimens was developed. The assay is based on the principle that upon non-denaturing polyacrylamide gel electrophoresis RNA molecules separate into several metastable conformational forms. These conformational polymorphisms strictly depend on the nucleotide sequence of the individual molecule. Using DNA from formalin-fixed, paraffin-embedded tissue of patients with mycosis fungoides, highly variable junctional sequences of rearranged T-cell receptor gamma genes were amplified by polymerase chain reaction. Subsequently, the polymerase chain reactions products were transcribed into complementary RNA and analyzed by non-denaturing polyacrylamide gel electrophoresis. In clinical specimens with a monoclonal lymphoid population, a clone-specific pattern of bands was identified representing conformational polymorphisms of cRNA molecules of rearranged T-cell receptor gamma genes of the predominant lymphoid clone. Three biopsies from one patient taken from different sites of the body over 3 years yielded an identical pattern of bands. This methodology provides a novel and rapid tool for the molecular identification and characterization of clonal lymphoid populations in clinical specimens. It is likely to be of special value for studies on the clonal evolution of lymphoid disorders of the skin. PMID- 8409517 TI - Effects of sunscreens and a DNA excision repair enzyme on ultraviolet radiation induced inflammation, immune suppression, and cyclobutane pyrimidine dimer formation in mice. AB - Exposure of skin to ultraviolet (UV) radiation inhibits the induction of delayed type hypersensitivity (DTH) responses initiated at a distant, unirradiated site. Recent studies attributed this form of immune suppression to DNA damage in the form of cyclobutane pyrimidine dimers (CPD). In the present study, we investigated the protective defects of sunscreens on UV-induced systemic suppression of DTH to Candida albicans, inflammation, and DNA damage. The photoprotective effects of sunscreen preparations containing 8% octyl-N-dimethyl p-aminobenzoate, 7.5% 2-ethylhexyl-p-methoxycinnamate, or 6% benzophenone-3 were studied in C3H mice exposed to a single dose of 500 mJ/cm2 UVB radiation from FS40 sunlamps. Inflammation was determined by the amount of skin edema at the site of UV irradiation, and DNA damage was assessed by measuring the frequency of endonuclease-sensitive sites in the epidermis. Application of the sunscreens before UV irradiation gave 75-97% protection against UV-induced edema, 67-91% protection against formation of CPD, but only 30-54% protection against suppression of DTH. In contrast, the topical application of liposomes containing a CPD-specific DNA repair enzyme immediately after UV irradiation resulted in 82% protection against suppression of DTH, but at best, 39% protection against skin edema. These findings demonstrate that sunscreens give less protection against UV induced immune suppression than against skin edema and CPD formation. Furthermore, they suggest that less DNA damage is required to cause UV-induced immune suppression than to cause sunburn. PMID- 8409518 TI - Analysis of the mechanism of ultraviolet (UV) B radiation-induced prostaglandin E2 synthesis by human epidermoid carcinoma cells. AB - Stimulation of cultured human keratinocytes with interleukin (IL)-1 alpha is known to elicit prostaglandin (PG) E2 release. Ultraviolet (UV) B radiation induces keratinocyte PGE2 and cytokine production. The present study deals with the autocrine roles of UVB-induced, keratinocyte-derived cytokines IL-1 and tumor necrosis-factor (TNF) alpha and their corresponding receptor molecules for UVB induced PGE2 release. In vitro exposure of transformed human keratinocytes (KB cells) induced PGE2 production five- to eightfold. This increase was inhibited by 70%, if irradiated cells were cultured in presence of monoclonal antibody (MoAb) M4, which blocks IL-1 effects by binding to the type 1 IL-1 receptor (IL-1R). In contrast, MoAb M22, which blocks the type 2 IL-1R, had no significant effects. Addition of recombinant human TNF alpha to unirradiated KB cells resulted in five to eightfold increased PGE2 synthesis, and this increase could be mimicked by stimulation of KB cells with MoAb htr-9, which exerts TNF alpha-like bioactivity by binding to the 55-kD TNF receptor (TNFR). UVB-induced PGE2 synthesis was blocked by 50% in the presence of neutralizing anti-TNF alpha-Ab, and was completely inhibited by addition of both anti-TNF alpha-Ab and MoAb M4. To elucidate a possible regulatory intracellular step in PGE2 synthesis, specific cyclooxygenase activity in KB cells was determined. Following UVB treatment, cyclooxygenase activity increased twofold, but remained unaltered, if irradiated KB cells were cultured in the presence of anti-TNF alpha-Ab plus MoAb M4. These studies indicate that keratinocyte-derived TNF alpha and IL-1 together mediate UVB-induced PGE2 release via specific cell surface receptors, and that one intracellular mechanism is an increased prostanoid-synthesizing capacity of irradiated cells. PMID- 8409519 TI - Activation of neutrophil membrane-associated oxidative metabolism by ultraviolet radiation. AB - Exposure of human neutrophils to ultraviolet radiation (UVR) in vitro was accompanied by activation of superoxide generation and preferential release of secondary granules. These pro-oxidative interactions of UVR with neutrophils were dependent on intact cellular membrane-associated oxidative metabolism and were mediated almost exclusively by the UVB component of UVR. Irradiation of neutrophils was also associated with release of arachidonic acid from membrane phospholipids, implicating involvement of phospholipase A2 (PLA2) in the pro oxidative activity of UVR. The pro-oxidative interactions of UVR with neutrophils were mimicked by coincubation of the cells with reagent arachidonate or lysophosphatidylcholine (LPC), whereas the PLA2 inhibitor 4-p-bromophenacyl bromide, as well as the LPC- and arachidonate complex-forming agent alpha tocopherol, inhibited these pro-oxidative interactions of UVR with phagocytes. Because phagocyte-derived reactive oxidants are cytotoxic, immunosuppressive, and carcinogenic, these agents are potential mediators of UVR-mediated tissue damage and tumorigenesis. PMID- 8409520 TI - Profiles of cytokine mRNA expressed by dendritic epidermal T cells in mice. AB - The epidermis of mice contains, in addition to Langerhans cells, a second dendritic population that is Thy-1+/CD3+/CD4-/CD8-/T-cell receptor-V gamma 3/V delta 1+. These dendritic epidermal T cells (DETC) are now thought to comprise one element in the family of epithelial tissue-resident gamma delta T cells. In the present study, DETCs were examined for their expression of mRNA for cytokines, using a reverse transcription-polymerase chain reaction. Freshly isolated Thy-1+ epidermal cells constitutively expressed mRNA for gamma interferon, but not IL-2. Within 24 h after stimulation with Con A, these cells then expressed mRNA for gamma-interferon and IL-2, but not IL-4. The rapid onset of expression of mRNA for IL-2 occurred exclusively within the Thy-1+ population, and in a Con A-dependent fashion. When freshly isolated epidermal cells were first stimulated with Con A and then expanded in bulk with rIL-2 for 20-24 d, cells expressing IL-4 mRNA then emerged, upon secondary stimulation with Con A. These "short-term" DETC lines also expressed mRNA for IL-2, interferon-gamma, IL 1 alpha, IL-3, IL-6, IL-7, tumor necrosis factor alpha and beta, and granulocyte macrophage-colony stimulating factor. Interestingly, mRNA for IL-4 and IL-6 was no longer detected in long-term (> 1 year) DETC lines 7-17 and 12-12. In addition, one line (7-17) maintained IL-3 mRNA expression, whereas another (12 12) had lost this capacity. These results emphasize the concept that, as resident cells in epidermis, DETCs exhibit several different immunorelevant activities, and the heterogeneity in cytokine mRNA profiles suggests that DETCs may divide into functional subsets. PMID- 8409521 TI - Reciprocal cytokine-mediated cellular interactions in mouse epidermis: promotion of gamma delta T-cell growth by IL-7 and TNF alpha and inhibition of keratinocyte growth by gamma IFN. AB - A unique subset of gamma delta T cells, termed dendritic epidermal T cells (DETC), resides in symbiosis with keratinocytes in mouse epidermis. We have shown previously that interleukin 7 (IL-7) which is produced by keratinocytes, promotes growth and prevents apoptosis in DETC. To extend this observation, we examined 12 cytokines, each of which is expressed by epidermal cells at mRNA and/or protein levels, for their capacities to modulate the growth of DETC. Cytokines examined included IL-1 alpha, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-10, tumor necrosis factor-alpha (TNF alpha), interferon-gamma (IFN gamma), granulocyte/macrophage colony stimulating factor (GM-CSF), and macrophage inflammatory protein-1 alpha (MIP-1 alpha). When tested individually, IL-2 and IL-7 promoted maximal growth of the long-term cultured DETC line 7-17. When tested in combinations, synergistic growth-promoting effects were seen with IL-2 and IL-4 or IL-7, and with IL-7 and IL-4 or TNF alpha. Dose-response experiments demonstrated that TNF alpha, which is produced by keratinocytes, enhances IL-7-induced DETC proliferation, but inhibits IL-2-induced proliferation. The mouse keratinocyte-derived cell line Pam 212 was used to test these cytokines for their capacities to regulate keratinocyte growth. Only gamma IFN, which is produced by DETC, inhibited proliferation in a dose-dependent fashion. These results illustrate three reciprocal pathways by which epidermal cytokines regulate the growth of epidermal cells: 1) a paracrine mechanism by which keratinocyte-derived cytokines (e.g., IL 7 and TNF alpha) promote the growth of DETC, 2) an autocrine mechanism by which DETC-derived cytokines (e.g., IL-2 and IL-4) support their own growth, and 3) a reciprocal pathway in which a cytokine produced by resident epidermal leukocytes (e.g., gamma IFN) modulates the growth of keratinocytes. PMID- 8409522 TI - Pentoxifylline suppresses irritant and contact hypersensitivity reactions. AB - Pharmacologic suppression of the effector phase of contact hypersensitivity appears to have major relevance with regard to treatment of type IV reactions like contact dermatitis. Recently, tumor necrosis factor alpha has been shown to be a critical mediator in hapten-induced irritant and contact hypersensitivity reactions, thus offering new possibilities, for therapeutic intervention. Pentoxifylline, a methylxanthine derivative used in the treatment of vascular disorders, currently has been found to suppress the production of tumor necrosis factor alpha by human and murine leukocytes. Therefore, the effect of pentoxifylline on the elicitation phase of contact hypersensitivity was studied. Intraperitoneal injection of pentoxifylline into sensitized Balb/c and C3H/HeN mice before application of the challenging hapten dose resulted in a significant reduction of the outcome of the contact hypersensitivity reaction. The suppressive effect of pentoxifylline was dose dependent and maximally pronounced upon injection 3 h before hapten application. In contrast to the effector phase of contact hypersensitivity, induction of contact hypersensitivity was not affected by pentoxifylline when injected into naive mice before performance of sensitization. In addition, irritant dermatitis induced by 1% croton oil or 5% benzalkonium chloride was suppressed by pentoxifylline as well. These data suggest a potential pharmacologic intervention, with pentoxifylline as a means to treat contact dermatitis. PMID- 8409523 TI - Differential expression of protein kinase C isoenzymes in normal and psoriatic adult human skin: reduced expression of protein kinase C-beta II in psoriasis. AB - Psoriatic lesions contain elevated levels of 1,2-diacylglycerol, the physiologic activator of protein kinase C (PKC), suggesting that PKC activation may be aberrant in psoriasis. We therefore have investigated the expression and properties of PKC isozymes in normal and psoriatic skin and in human skin cells. Chromatographic and immunoblot analyses revealed the presence of the calcium dependent PKC isozymes PKC-alpha and -beta, but not -gamma, in normal human epidermis. PKC-beta was more prominent, constituting two thirds of the total calcium-dependent PKC activity. In psoriatic lesions, expression of both PKC alpha and -beta was decreased, with preferential reduction (80%) of PKC-beta. Northern analysis and semi-quantitative polymerase chain reaction (PCR) indicated no change in the mRNA levels of PKC-alpha and -beta between normal and psoriatic epidermis. In normal epidermis, PKC-alpha was expressed mainly in the lower epidermis, whereas PKC-beta was localized to the upper cell layers, with very intense staining of CD1a+ Langerhans cells. In psoriasis, PKC-alpha staining was present in the lower epidermis, whereas PKC-beta staining was essentially absent, with the exception of some positive inflammatory cells. In addition to PKC-alpha and beta, immunoblot and Northern/PCR analysis revealed expression of four calcium-independent PKC isozymes, delta, epsilon, zeta, and eta, in both normal and psoriatic skin. There were no significant differences in mRNA levels among any of these PKC isozymes, between normal and psoriatic skin. Soluble PKC-zeta protein was modestly increased (twofold) in psoriatic, compared to normal, skin, whereas the levels of PKC-delta, epsilon, and eta were unchanged. Analysis of PKC isozyme expression in the three major cell types of human epidermis revealed that Langerhans cells and keratinocytes were the major sources of PKC-beta and PKC zeta, respectively. These data demonstrate the diversity of PKC isozyme expression in human skin, and suggest that alterations of PKC-beta and -zeta may participate in the aberrant regulation of growth and differentiation observed in psoriasis. PMID- 8409524 TI - Evidence for an altered protein kinase C (PKC) signaling pathway in psoriasis. AB - Treatment of normal primary human keratinocytes with phorbol 12-myristate 13 acetate (PMA) or phorbol 12-13 dibutyrate (PDBu) (100 ng/ml, 6-40 h) followed by two-dimensional (2-D) gel electrophoresis (isoelectric focusing) and microsequencing identified three polypeptides (phorbolin 1, M(r) = 19.9 kDa; phorbolin 2, M(r) = 19.7 kDa; and interleukin-1 (IL-1) receptor antagonist, IL 1ra, M(r) = 19.5 kDa) that are upregulated eight times or more by the phorbol esters and that are highly expressed in noncultured psoriatic keratinocytes. The response was not elicited by other effectors tested including second messengers (Bt2cAMP, Bt2cGMP), cytokines (basic fibroblast growth factor, transforming growth factor-alpha, IGF-II, tumor necrosis factor-alpha, and -beta, interleukin (IL)-1 alpha, IL-1 beta, IL-2, IL-3, IL-6, IL-7, IL-8, interferon-alpha, and gamma), and other substances (Ca++, dexametasone, retinoic acid, lipopolysaccharides) and it was partially reversed by staurosporine, a strong inhibitor of protein kinase C. The results are taken to imply that the protein kinase C signaling pathway may be altered in psoriatic keratinocytes. PMID- 8409525 TI - Phorbol ester TPA- and bradykinin-induced arachidonic acid release from keratinocytes is catalyzed by a cytosolic phospholipase A2 (cPLA2). AB - In a previous paper, we have shown that bradykinin (Bk) and the phorbol ester 12 O-tetradecanoylphorbol-13-acetate (TPA) stimulate arachidonic acid release from HEL-30 keratinocytes along a Bk-B2 receptor G-protein-coupled pathway or a protein kinase C-dependent mechanism, respectively. Here we show a cytosolic PLA2 (cPLA2) to be responsible for this effect. This enzyme exhibited a marked acyl group specificity towards arachidonic acid. It was activated by Ca++ in micromolar concentrations and partially translocated from the cytoplasmic to the membrane fraction upon Ca++ treatment. Translocation was also observed upon treatment of cells with either Bk or TPA. However, only with Bk was a corresponding increase of the cytoplasmic Ca++ level observed, whereas TPA induced translocation occurred at basal Ca++ concentrations. Indirect evidence for a G protein to be involved in Bk- but not TPA-dependent cPLA2 activation was provided using non-hydrolyzable GTP derivatives. It is concluded that keratinocyte cPLA2 plays a critical role in the initiation by exogenous and endogenous factors of the eicosanoid cascade in skin. PMID- 8409526 TI - Time- and stimulus-dependent secretion of NAP-1/IL-8 by human fibroblasts and endothelial cells. AB - The neutrophil-activating peptide 1/interleukin 8 (NAP-1/IL-8) has in the past been extensively characterized biochemically as well as functionally. Effects of NAP-1/IL-8 on inflammatory cells like neutrophilic granulocytes and lymphocytes, as well as its production by several different cell types, point towards an important role in different inflammatory processes. Recently, monoclonal antibodies have helped to establish immunoassays for detecting the peptide. Using such antibodies, we have performed in vitro studies on the time- and stimulus dependent production of IL-8 by endothelial cells as well as fibroblasts. Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 alpha (IL-1 alpha) efficiently induced both focal intracellular expression as well as secretion of the peptide when tested by immunocytochemistry and enzyme-linked immunosorbent assay (ELISA). After stimulation with phorbol myristate acetate (PMA) and lipopolysaccharide (LPS), such effects were seen only in endothelial cells, whereas interferon (IFN)-gamma did not induce any pronounced effect on either of the cells tested. These studies demonstrated in vitro release of IL-8 by different cells upon specific stimulation, thus underlining the significance of the in vivo secretion of this peptide, as noted in recent studies. PMID- 8409527 TI - Induction of hair growth in ear wounds by cultured dermal papilla cells. AB - In the adult hair follicle the dermal papilla plays a crucial role in the dermal epidermal interactions that control hair production and events of the growth cycle. It has previously been shown that cultured cells from rat vibrissa follicle dermal papillae can stimulate hair growth when implanted into amputated follicles. This study investigated the effects of implanting low-passage cultured papilla cells into small incisional wounds in the rat ear pinna. The groups of fibers that emerged from wound sites were much larger than local hairs, and often had vibrissa-type characteristics. Later-passage papilla cells or cultured skin fibroblasts failed to elicit the same response. Histology revealed that big follicles were formed when papilla cells were trapped between the cut edges of the epidermis. Abnormally large follicles were seen at wound sites many months post-operatively. Independent of epidermal influence, cultured papilla cells in the wound dermis formed rounded papilla-like aggregates that also persisted until biopsy. A previously described method of wrapping papilla cells in glabrous epidermis was less successful in percentage terms but resulted in the production of one very large vibrissa-type follicle and fiber. These results further illustrate that the inductive powers and developmental information retained by cultured dermal papilla cells parallel the properties of their embryonic precursors; the findings may have implications for human hair growth. PMID- 8409528 TI - Mouse skin is particularly susceptible to tumor initiation during early anagen of the hair cycle: possible involvement of hair follicle stem cells. AB - Stem cells are believed to be a necessary target of chemical carcinogens. Based on autoradiographic, ultrastructural, and biologic criteria, we have recently proposed that hair follicle stem cells reside not in the bulb, but in the upper outer root sheath in an area called the bulge. Proliferating cells have been shown to be more susceptible to tumor initiation, and we have recently demonstrated that cells in the bulge undergo transient proliferation during early anagen. Therefore, we theorized that mouse skin should be particularly susceptible to carcinogen application during early anagen phase. In this paper, we show that early anagen Swiss and Sencar mouse skin is indeed particularly susceptible to one- and two-stage chemical carcinogenesis, resulting in tumor yields one to five times those obtained with telogen-timed carcinogen application. Our findings implicate a possible involvement of the bulge cells as precursors to some of the skin cancers, and support the concept that these are stem cells. These observations also raise important questions about the cellular origins and biologic behavior of chemically induced murine skin tumors. PMID- 8409529 TI - 7,12-Dimethylbenz[a]anthracene-induced mouse keratinocyte malignant transformation independent of Harvey ras activation. AB - Independent clones of mouse keratinocytes initiated in vitro gave rise to tumor phenotypes typical of mouse skin multistage carcinogenesis and histologically similar to human tumors of the skin, and head and neck. High-molecular-weight genomic DNAs isolated from two 7,12-dimethylbenz[a]anthracene (DMBA)-initiated murine epithelial carcinoma cell lines and one papilloma cell line were examined for transforming activity by transfection into NIH3T3 cells. DNAs from each of these cell lines resulted in the formation of foci morphologically unlike foci containing an activated c-Ha-ras oncogene. Following polymerase chain reaction amplification of the c-Ha-ras gene, Xba I restriction analysis and oligonucleotide differential hybridization did not detect 61st, 12th, or 13th codon mutations. Southern and Northern analysis confirmed that the normal c-Ha ras gene was not activated by amplification or overexpression. These results provide evidence that 7,12-dimethylbenz[a]anthracene-induced malignant transformation of murine keratinocytes occurred independent of point mutations associated with c-Ha-ras activation. The absence of an activated c-Ha-ras oncogene in these cell lines distinguishes our model from other mouse models of carcinogenesis and may provide a model for functional genetic changes during initiation and progression of human epithelial cancers. PMID- 8409530 TI - Distinctive integrin expression in the newly forming epidermis during wound healing in humans. AB - The integrin receptor family plays a fundamental role in mediating cell attachment to a variety of extracellular matrix molecules. In normal human epidermis, the alpha 2 beta 1, alpha 3 beta 1, alpha 6 beta 4, and alpha v beta 5 integrin heterodimers are expressed and appear largely confined to the basal cell layer. In the present study, beta 1, beta 4, and alpha v integrin expression in the epidermis during wound healing in humans was examined. Punch biopsies were performed on healthy volunteers. At daily intervals up to day 8, and at days 11, 14, 21, and 28, the wound site was surgically removed. Using immunofluorescence microscopy, several modifications of the integrin expression pattern were observed on migrating keratinocytes during the re-epithelialization phase of the wound-healing process: i) alpha v expression was strongly enhanced and polarized at the basal pole of basal keratinocytes; ii) among the beta 1 integrins, alpha 3 beta 1 was overexpressed and distributed over the entire basal keratinocyte membrane and a weak alpha 5 beta 1 reactivity became evident; and iii) alpha 6 beta 4 was detected as a linear staining along the newly forming dermal-epidermal junction. Moreover, both during the re-epithelialization phase and during the first 2 weeks after wound closure, alpha 3, alpha 6, alpha v, beta 1, and beta 4 were no longer confined to the basal layer, as in normal epidermis, but were also found on several suprabasal cell layers. These results suggest that alpha v beta 5, alpha 3 beta 1, and alpha 5 beta 1 may be the main integrin receptors mediating keratinocyte spreading and migration over the provisional matrix of the wound bed. PMID- 8409531 TI - Melanocyte movement in vitro: role of matrix proteins and integrin receptors. AB - During the repigmentation of vitiliginous skin, melanocytes migrate from the outer root sheath of the hair follicle into the depigmented skin. We hypothesize that this requires changes in the local microenvironment that are conductive to melanocyte migration. One important change in the microenvironment could be the localized production of matrix proteins. We have previously employed time-lapse photography to evaluate the effect of inflammatory mediators and cytokines on melanocyte movement. We have adapted this system to study the effect of matrix proteins on melanocyte movement in vitro. Type IV collagen significantly increases melanocyte migration, whereas laminin and fibronectin have no effect. Cell/matrix interactions are in part controlled by cell-surface integrins. Integrins have been demonstrated to be important in controlling the migration of many cell types. We demonstrate that melanocytes express cell-membrane alpha 2, alpha 3, and alpha 5 integrins and that the enhanced melanocyte migration on type IV collagen is inhibited by specific function-blocking antibodies to integrins alpha 2 and alpha 3, but not to alpha 5 integrins. PMID- 8409532 TI - Effect of organic solvents on in vitro human skin water barrier function. AB - Skin barrier disruption caused by organic solvents to human cadaver dermatomed skin was evaluated using an in vitro model system. Resultant changes in transepidermal water loss (TEWL), as measured with an evaporimeter, were recorded after topical application of either acetone, chloroform:methanol 2:1, hexane, hexane:methanol 2:3, or the control, water, for exposure times of 1, 3, 6, and 12 min. The resultant lipid/solvent mixture was removed and analyzed for its lipid content. The ability of the different solvents to induce changes in the skin's barrier function was assessed by comparing pre- to post-solvent exposure TEWL (delta TEWL). When compared to the controls, water and unexposed skin, chloroform:methanol 2:1 caused the greatest significant increase in TEWL, followed by hexane:methanol 2:3. Acetone and hexane showed no difference in TEWL from the controls. Besides solvent, exposure time was a significant independent variable for predicting delta TEWL, and the interaction of the two (exposure time and solvent type together) was the strongest predictor. Lipid analysis of the extracts revealed that all the solvents removed comparable quantities of the surface lipids (triglycerides, wax esters, squalene, cholesterol esters). Stratum lipids--ceramides, free fatty acids, and cholesterol--extracted by chloroform:methanol 2:1 and hexane:methanol 2:3 were comparable and significantly greater than those extracted by acetone and hexane. These two solvents failed, however, to induce comparable changes in TEWL, as chloroform:methanol 2:1 induced a significantly greater delta TEWL than hexane:methanol 2:3. Additionally, no individual lipid class extracted by either chloroform:methanol 2:1 or hexane:methanol 2:3 proved to be a significant or accurate variable for predicting delta TEWL. This suggests that the mechanism by which topical chloroform:methanol 2:1 and hexane:methanol 2:3 exposure induce a delta TEWL involves more than pure lipid extraction. PMID- 8409533 TI - Influence of schedule and mode of administration on effectiveness of podofilox treatment of papillomas. AB - We investigated the timing of podofilox delivery to see how it correlated with papilloma size. We looked at times ranging from once per week (morning and afternoon) to five times per week (morning and afternoon). We also looked at delivery systems that might enhance the effectiveness of the drug by increasing penetration of the overlying cutaneous horn. These included soaking prior to drug administration, the use of a grooved needle subsequent to drug administration, and the various possible combinations of these techniques. We found that the timing of treatments had relatively little effect on the size of the papillomas. For example, at the end of ten weeks, the geometric mean diameter (GMD) (mm) of the papillomas treated five times per week and induced by a 10(-1) dilution of the virus (group B) was 18. Likewise the GMD (mm) of papillomas treated once per week was 18 (group D). On the other hand, we found that soaking plus the use of a grooved needle plus podofilox resulted in the curing of all lesions induced by a 10(-2) virus dilution and of most induced by the 10(-1) dilution, whereas soak plus podofilox or podofilox alone were not as effective in curing the lesions. Podofilox plus needle approached the soak plus podofilox plus needle in effectiveness. Treatment schedule was not a critical determinant of podofilox effectiveness. Therapeutic benefits were enhanced by hydration of the overlying cutaneous horn and penetration with a grooved needle. PMID- 8409534 TI - Identification of a basement membrane zone antigen reactive with circulating IgA antibody in ocular cicatricial pemphigoid. AB - Ocular cicatricial pemphigoid is a rare vesiculobullous disease characterized by linear deposition of IgG and/or IgA along the basement membrane zone of conjunctival biopsies. This study identifies a tissue antigen detected by ocular cicatricial pemphigoid patient sera. Patient selection was based on the presence of only ocular involvement and a positive direct immunofluorescence of conjunctiva. We evaluated patient and control sera using indirect immunofluorescence of basement membrane zone separated skin, Western blot, and purified antibodies from nitrocellulose and epidermal sheets. Direct immunofluorescence performed on the patients' conjunctival biopsy showed linear deposition of IgA along the basement membrane zone in all seven patients, and five of seven also demonstrated deposition of IgG along the basement membrane zone. Indirect immunofluorescence performed on the patients' sera demonstrated linear deposition of IgA along the epidermal side of the basement membrane zone of ethylenediaminetetraacetic acid-separated skin in all seven patients. IgA titers ranged from 1:20 to 1:80. No IgG was detected. Immunoblots detected IgA binding to a 45-kD antigen in all patients as well as sporadic IgA binding to a number of other proteins. Immunoblots stained with sera from patients did not show reactivity to the 230- or 180-kD bullous pemphigoid antigens or the 97-kD linear IgA bullous dermatosis antigen. Eluting IgA from the 45-kD region and other regions revealed that only antibodies eluted from the 45-kilodalton region bound linearly to the basement membrane on separated skin. Purification of IgA using epidermal sheets confirmed that the antibody responsible for staining on indirect immunofluorescence bound to the 45-kD region on Western blot. Sera from normals and patients with bullous pemphigoid, dermatitis herpetiformis, and linear IgA bullous dermatosis failed to demonstrate basement membrane zone IgA on elution of the 45-kD region. We conclude that these ocular cicatricial pemphigoid sera contain a unique IgA antibody that binds to a 45-kD basement membrane zone antigen. PMID- 8409535 TI - Skin explant culture: a reliable method for detecting pemphigoid antibodies in pemphigoid sera that are negative by standard immunofluorescence and immunoblotting. AB - We investigated the presence of bullous pemphigoid antibodies in bullous pemphigoid sera that are negative by standard indirect immunofluorescence. We incubated each of four indirect immunofluorescence-positive bullous pemphigoid sera, seven indirect immunofluorescence-negative bullous pemphigoid sera, one indirect immunofluorescence-negative herpes gestationis serum, three indirect immunofluorescence-positive epidermolysis bullosa acquisita sera, five indirect immunofluorescence-negative epidermolysis bullosa acquisita sera, and two normal human sera with fresh human skin explants in medium 199 at 4 degrees C for 48 h. All bullous pemphigoid sera, herpes gestations serum, and the three indirect immunofluorescence-positive epidermolysis bullosa acquisita sera had IgG that bound the basement membrane zone of skin explants with moderate to marked intensity as demonstrated by immunofluorescence. Normal sera and indirect immunofluorescence-negative epidermolysis bullosa acquisita sera failed to bind the explant basement membrane zone. Immunoblotting of bullous pemphigoid sera showed five of seven indirect immunofluorescence-negative bullous pemphigoid sera to bind high-molecular weight and/or low-molecular weight bullous pemphigoid antigens from epidermal extracts. We conclude that the skin explant culture system is a very sensitive method for the detection of bullous pemphigoid antibodies in all bullous pemphigoid sera. PMID- 8409536 TI - Altered proliferation, synthetic activity, and differentiation of cultured human sebocytes in the absence of vitamin A and their modulation by synthetic retinoids. AB - Human sebocytes maintained in medium containing delipidized serum were studied for ultrastructural characteristics, cell proliferation, lipid synthesis, immunophenotype, and keratin expression before and after the addition of the synthetic retinoids isotretinoin and acitretin (10(-8)-10(-5) M). Compared to the properties of sebocytes cultured in normal sebocyte medium (1-2 x 10(-7) M vitamin A), the use of delipidized serum (undetectable amounts of vitamin A) resulted in prominent decrease of i) proliferation; ii) number of intracellular lipid droplets and synthesis of total lipids, especially triglycerides, squalene, and wax esters; and iii) labeling with monoclonal antibodies identifying progressive and late-stage sebocyte differentiation. Intercellular spaces narrowed and cell-to-cell contacts were established by abundant desmosomes. Lanosterol was induced. Keratins 14, 16, 17, and 18 were upregulated and the keratin 16: keratin 4 ratio, negatively correlating with sebocyte differentiation, increased. Addition of isotretinoin and acitretin exerted a biphasic effect. At concentrations < or = 10(-7) M, both compounds enhanced sebocyte proliferation and synthesis of total lipids, especially triglycerides and cholesterol, and decreased lanosterol, keratin 16, and the keratin 16:keratin 4 ratio. In contrast, retinoid concentrations > 10(-7) M inhibited sebocyte proliferation in a dose-dependent manner. Our findings indicate that vitamin A is essential for proliferation, synthetic activity, and differentiation of human sebocytes in vitro. Synthetic retinoids partially reinstate the altered functions of sebocytes maintained in medium containing delipidized serum. In contrast to the previously shown isotretinoin-specific response of cultured sebocytes in the presence of vitamin A, similar effects of isotretinoin and acitretin were obtained in its absence. This suggests different interactions of synthetic retinoids with vitamin A, possibly influencing their efficacy on the sebaceous gland. PMID- 8409537 TI - Human hair follicle germinative epidermal cell culture. AB - Isolated human hair follicle germinative epidermal cells were observed in vitro for the first time. When cultured alone, this small, round, novel cell type did not grow, divide, take on an outer root sheath-type appearance, or display any obvious signs of epidermal differentiation. We have previously described comparable cells from rat vibrissa follicles. However, in combination with human hair follicle dermal papilla populations, the germinative epidermal cells were stimulated into proliferative and complex interactive behaviors. This included the formation of composite organotypic structures containing not only impressively intact basement membrane, but also the hair-specific form, glassy membrane. PMID- 8409538 TI - Comparison of approaches in the assessment of myocardial viability and follow-up interventional procedures. PMID- 8409539 TI - Comparison of approaches in the assessment of myocardial viability and follow-up of PTCA/CABG. The role of echocardiography. AB - Dysfunctional myocardium may be viable in patients with acute myocardial infarction. Although viable but dyssynergic myocardium may be recognized a posteriori by the occurrence of functional recovery, prospective identification of the actual myocardial state is more important for optimal therapeutic management. Echocardiography during pharmacological interventions is a useful clinical tool, as changes in regional myocardial thickening may be continuously monitored. Stunned myocardium exhibits contractile dysfunction after an ischemic episode despite normalized or near normalized myocardial flow, but stunned myocardium retains contractile reserve. It may be identified by improvement in contractility during a low dose dobutamine infusion in segments showing a mismatch between normal perfusion and reduced contractility. In regions of viable but stunned myocardium corresponding to an artery with reduced coronary flow reserve, contractility may improve at low dose dobutamine infusion and may later deteriorate at high dose, indicating the presence of jeopardized myocardium. Ischemia at a distance observed with dobutamine indicates the presence of multivessel coronary artery disease. These informations are useful for clinical decision making. PMID- 8409540 TI - Non-invasive assessment of residual viability in post-myocardial infarction patients. Role of nuclear techniques. AB - The failure of non-imaging techniques in identifying viable segments has favoured the clinical application of nuclear imaging. The main pathways that support radionuclide imaging are cell membrane integrity, persistence of intermediary metabolism and demonstration of a residual coronary reserve. Thallium-201 reinjection or rest protocols allow the identification of viable myocardium in most of patients with wall motion abnormalities and appear to be the most diffuse, low-cost and available method to detect viable myocardium. More complex approaches use positron emission tomography and matched flow/metabolic information. Flow/metabolic 'mismatch' usually identifies most of hypoperfused regions that show post-operative improvement of regional wall motion. The last promising approach is represented by the demonstration of a maintained regional coronary reserve in dyssynergic areas. Technetium-99m-microspheres (or Teboroxime in the future) can be successfully used for this purpose. The clinical application of radionuclides appears to be one of the principal imaging tools able to identify residual viability. PMID- 8409541 TI - Assessment of viability after myocardial infarction. Clinical relevance and methodological problems. AB - In patients with myocardial infarction, the distinction between reversible and irreversible ventricular dysfunction has important clinical implications since dysfunctional but viable myocardium will resume contraction following revascularization. Various methods have been developed for the identification of potentially reversible myocardial dysfunction. Thallium reinjection, immediately after stress-redistribution imaging, may provide evidence of myocardial viability by demonstrating thallium uptake in regions with apparently 'irreversible' defects. Hypoperfused, hypocontractile segments may recover function after revascularization, when exhibiting increased 18F-fluoro-deoxy-glucose uptake on positron emission tomography. Improved contractile function by selective beta 1 adrenergic stimulation with low dose dobutamine may also indicate the presence of viable tissue and predict subsequent improvement upon restoration of adequate flow. Finally, exercise-induced ST segment elevation on leads exploring a recent myocardial infarction has also been shown to indicate the presence of viable, potentially salvageable tissue. We discuss here these and several other methods that have been proposed for the detection of residual myocardial viability. Their advantages, limitations, and relevance to clinical problems are also discussed. PMID- 8409542 TI - Assessment of residual viability in patients with myocardial infarction using magnetic resonance techniques. AB - Magnetic resonance techniques have only recently been employed to assess residual myocardial viability after myocardial infarction. Three approaches have been described to achieve this purpose: First, the use of signal intensity changes on spin-echo images with and without the application of contrast media to define irreversible injury to the myocardium in acute and subacute infarcts; second, measurement of metabolite concentrations within the infarct area using magnetic resonance spectroscopy, and third, quantitation of myocardial thickness and systolic wall thickening in chronic infarcts. This paper reviews the pertinent literature and compares MR techniques with other imaging techniques used in the diagnosis of myocardial viability. PMID- 8409543 TI - Value and limitations of exercise stress testing to predict the functional results of coronary artery bypass grafting. AB - To assess the value of exercise stress testing to predict the functional result of revascularization, 90 patients were evaluated by coronary angiography and exercise testing pre and postoperatively. Patients were classified on the basis of the postoperative angiogram in a group with successful surgery and a group with unsuccessful surgery. The predictive accuracy positive of ST segment depression to detect unsuccessful surgery was 67% The predictive accuracy negative was 61%. The best predictor of unsuccessful surgery was residual angina pectoris after revascularization with predictive value positive and negative of 85% and 60%, respectively. Thus exercise stress testing has limited value to accurately predict the degree of revascularization. PMID- 8409544 TI - The role of scintigraphic techniques in the evaluation of functional results of coronary bypass grafting and percutaneous transluminal coronary angioplasty. AB - Scintigraphic techniques can be used first, to guide appropriate referral for interventional procedures, and second to predict the effect of revascularization on regional perfusion and function prior to the intervention, thereby being able to assess efficacy of revascularization and to assess whether ischemia is the origin of recurrence of symptoms. Of increasing importance is the ability of nuclear techniques to identify those myocardial regions with abnormal function which might benefit from revascularization by showing improvement in regional wall motion. Positron emission tomography is considered to be the gold standard to assess regional myocardial perfusion and metabolism. The introduction of the reinjection technique makes 201Tl-scintigraphy the method of choice to detect jeopardized myocardium and to guide appropriate referral for revascularization procedures in those institutes where PET is not available. Even when the costly PET-instrumentation is available, cost-benefit analysis is indicated to assess the additional value of PET compared with 201Tl reinjection imaging. PMID- 8409545 TI - The role of magnetic resonance in the evaluation of functional results after CABG/PTCA. AB - Magnetic resonance imaging (MRI) is a non-invasive modality which can be used for direct visualization of coronary artery bypass grafts. Spin-echo and gradient echo (cine-MRI) techniques are now available on standard MR machines and provide information on graft morphology and graft patency with a 90% accuracy. By combining the standard techniques with MR phase velocity mapping, the flow rate in the graft can be measured, thereby offering a unique non-invasive assessment of the graft function. Newer techniques include MR coronary angiography, pharmacologically induced stress MRI, ultrafast MRI of the first-pass (perfusion) of a paramagnetic contrast agent through the myocardium, and 31P MR spectroscopy of high-energy phosphate metabolism of the myocardium. All of these may develop into valuable diagnostic tools for the assessment of functional results after CABG or PTCA, but still require clinical validation. At present, MRI is a useful screening procedure for assessment of graft patency and function in post operative pain syndromes and in late graft occlusion or stenosis. PMID- 8409546 TI - Indications for routine heart-catheterization after CABG and PTCA. AB - Routine heart-catheterization after Coronary Artery Bypass Graft surgery (CABG) or Percutaneous Transluminal Coronary Angioplasty (PTCA) has been advocated to determine the change in bypass graft or dilated coronary artery and native coronary artery status, the effective disease remaining after CABG or PTCA and the relation between progression of disease, left ventricular function and symptomatology. Results of angiographic follow-up data after CABG and PTCA are presented and the practical implications are discussed. The reliability of symptoms, invasive and non-invasive test for the detection of ischemia are considered. Finally, recommendations are made for the indication of routine heart catheterization after CABG and PTCA. PMID- 8409547 TI - Clinical assessment following coronary revascularization. AB - There remains a need to establish adequate protocols for investigating the short- and long-term follow-up of revascularization procedures. For coronary angioplasty the most reliable basis for decision-making in managing patients is the symptomatology of the patient. For bypass surgery a protocol should be established to evaluate patients late, at 5 to 10 years following bypass surgery, in particular those with saphenous vein grafting, as graft and patient survival begins to fall after this period. Investigation after this may be too late for many patients who may already have several occluded grafts and poor left ventricular function, two of the most important prognostic factors post bypass surgery. The improvement and refinement of non-invasive investigations has led to a better understanding of the value and limitations of many of these tests, but it is particularly important that the limitations of many investigation are fully appreciated when they are used to influence clinical decisions. In this regard, a study comparing and integrating the predictive value of the persistence or return to symptoms, a positive non-invasive test, and a positive invasive test would surely prove invaluable. PMID- 8409548 TI - How has echo/Doppler influenced the practice of adult cardiology? PMID- 8409549 TI - Has echo/Doppler influenced the practice of paediatric cardiology? PMID- 8409550 TI - Echocardiography in the diagnosis of thoracic aortic pathology. AB - Transthoracic two-dimensional and Doppler echocardiography has been well established as a useful technique for evaluating many pathologic processes affecting the thoracic aorta. However, the distance of the aortic arch and descending thoracic aorta from the chest wall and the interposition of highly attenuating lung and highly reflective mediastinal structures between the transducer and the aorta present unavoidable limitations. Transesophageal echocardiography is a relatively new technology that overcomes many of the inherent limitations with transthoracic imaging. Complete echocardiographic evaluation of the entire thoracic aorta can now be achieved in nearly all patients. This article will review the continually expanding role of echocardiography in the evaluation of thoracic aortic pathology, including the dramatic impact of transesophageal imaging on the diagnosis of life-threatening disorders such as aortic dissection. PMID- 8409551 TI - Milestones in cardiac ultrasound: echoes from the past. History of cardiac ultrasound. AB - Although the antecedents of cardiac ultrasound can be traced back to the 1870s, it was in 1954 that Edler and Hertz published their milestone paper. M-mode echocardiography reached its peak in the early 1970s, when the fibre-optic recorder made the method clinically viable. It was not long before real-time two dimensional imaging was developed, however, and the invention of pulsed Doppler laid the foundation of duplex scanning. In 1985, colour flow imaging gave a fresh impetus to echocardiography. In parallel with the main developments, intravascular and transoesophageal scanning have gained clinical popularity within the last decade, together with techniques for the display of three dimensional images. Other innovations include contrast agents, pressure gradient measurement and promising methods for tissue characterisation. It seems that current techniques are safe, but this needs to be kept continuously under review. PMID- 8409552 TI - Assessment of pulmonary embolism. PMID- 8409553 TI - Role of cardiac ultrasound in the assessment of hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy is a complex disease characterized by a broad spectrum of morphologic and functional abnormalities and by a great variability in its natural history. Cardiac ultrasound has had a great impact on our understanding of hypertrophic cardiomyopathy and has become the main tool for diagnosis and patient management. This brief review will summarize the main applications of cardiac ultrasound in this disease. PMID- 8409554 TI - Echocardiography and coronary artery disease. AB - Echocardiography is playing an increasingly important role in the management of patients with coronary artery disease. With the addition of new digital technology and new technological advances, such as multiplane transesophageal echocardiography and intravascular ultrasound, there is every expectation that this use of cardiac ultrasound will grow even more rapidly in the near future. PMID- 8409555 TI - Echocardiographic assessment of myocardial viability: clinical applications and future directions. PMID- 8409556 TI - Echocardiography and heart failure. PMID- 8409557 TI - Evolving trends and future directions in echocardiography. PMID- 8409558 TI - Secondary management of the nose in the cleft patient. AB - The management of the cleft lip and palate patient usually requires multiple primary and secondary surgical procedures to correct the deformity. This paper presents surgical methods in the secondary management of the cleft nose and presents three case reports to help illustrate these methods. PMID- 8409559 TI - Computed tomography in differential diagnosis of temporomandibular joint disorders. AB - Computed tomography (CT) has great potential for imaging intra- and extracapsular hard-tissue abnormality of the temporomandibular joint (TMJ). CT is not the best method of imaging disk position and form. For differential diagnosis of TMJ disorders, CT is especially successful in bony lesions. The study includes 16 examples of TMJ hard-tissue abnormality. In the differential diagnosis of all cases, CT played a decisive role. PMID- 8409560 TI - Anterior segmental maxillary osteotomy. A 24-month follow-up. AB - Anterior segmental maxillary osteotomies were performed in 14 patients. Both the downfracture and the Wunderer methods were used. The patients were examined regularly until 24 months after surgery. No major complications occurred. Long term stability of the osteotomy was found to be acceptable except when used to correct deep overbites. In these cases, other methods should be considered. PMID- 8409561 TI - Assessment of inferior alveolar and lingual nerve disturbances after dentoalveolar surgery, and of recovery of sensitivity. AB - In a follow-up of 1107 dentoalveolar operations in the postcanine region, 24 (2.2%) temporary sensitivity disturbances of the inferior alveolar nerve and 16 (1.4%) of the lingual nerve were found. Permanent disturbances were not present. Complete recovery had occurred by 6 months in all cases. The incidence of temporary sensitivity disturbances depended on the different surgical interventions performed. For evaluation and follow-up purposes, a computer-aided pain and thermal sensitivity (PATH) tester was used. By PATH testing, spontaneous recovery can already be ascertained at the third or fourth postoperative month. PMID- 8409562 TI - Carcinoma of the oral cavity in patients over 75 years of age. AB - The results have been analyzed of a 15-year retrospective study on the presentation and survival of 46 patients aged > or = 75 years with oral carcinoma. All tumors were staged according to the UICC criteria. The presence of other malignant tumors in nine, premalignancy in five, and systemic disease in 15 patients was noted. A large subpopulation of elderly women, nonusers of alcohol and tobacco, usually with advanced disease, was found. In one-quarter of the patients who underwent surgery, a major resection with primary flap reconstruction was judged to be necessary. In general, this type of surgery was well tolerated. Three-year survival was 60%; women, systemically compromised patients, and patients with stage III and IV tumors did worse. A recurrence ratio of 59% was found after 5 years, recurrence usually appearing during the first year. The conversion rate was low (8.5%). PMID- 8409563 TI - Immediate reconstruction following maxillectomy: a new method. AB - A new method for immediate reconstruction of the maxilla after resection is described. The ipsilateral pedicled temporalis muscle is tunneled into the defect and sagittaly split into two layers. The inner layer is used to line the nasal side. An individually shaped titanium mesh, tightly filled with free autogenous corticocancellous bone, is fixed by titanium screws to the remnant of the zygoma and contralateral maxilla. The outer layer of the split muscle covers the reconstruction, the temporalis fascia forming the oral side. The method resulted in good cosmetic appearance and permitted the re-creation of a maxillary alveolar ridge suitable for endosseous implants or a simple prosthesis. PMID- 8409564 TI - Natural course of jaw lesions in patients with familial adenomatosis coli (Gardner's syndrome). AB - Twenty-three patients suffering from familial adenomatosis coli (FAC) were followed for an average of 7 years (0.11-16.11 years). In 22 patients, jaw lesions including osteomas and/or odontomas were present at the first examination. In 12 patients, some changes occurred; that is, the number and size of the lesions increased. These findings indicate that changes of jaw lesions in FAC patients occur gradually even in adulthood. PMID- 8409565 TI - Benign cartilaginous tumor of the gingiva. A case report. AB - Benign cartilaginous tumors of the intraoral soft tissues are rare and are typically seen in the tongue, buccal mucosa, or soft palate. They comprise a diverse group of lesions that are usually defined as cartilaginous choristomas or chondromas. The case reported was located on the palatal maxillary gingiva of the first premolar of a 56-year-old white woman. It appeared as a small firm tumor covered by normal mucosa, with the underlying bone intact on radiographic examination. Histologic examination of the excised lesion revealed a circumscribed mass of cartilaginous tissue with occasional cells presenting atypical nuclear features. Two years after initial presentation, the patient is free of recurrence. PMID- 8409566 TI - Myospherulosis of the lower lip. Report of a case. AB - A 19-year-old man with myospherulosis of the lower lip is described. The lesion was probably caused by petrolatum-based antibiotic ointment. PMID- 8409567 TI - Xanthomatous lesion of the mandible. Report of a case. AB - Xanthomatous lesions of the jawbone are rare and ill-defined. In the case presented, infiltration of the mandibular bone marrow by abundant foam cells was combined with extensive reactive bone formation. PMID- 8409568 TI - A double-blind comparative study of soluble aspirin and diclofenac dispersible in the control of postextraction pain after removal of impacted third molars. AB - The analgesic efficacy and patient acceptability of soluble aspirin and diclofenac dispersible were compared in patients with postoperative pain after removal of impacted third molars. A total of 136 patients were randomly allocated to receive soluble aspirin 600 mg tds or diclofenac dispersible 50 mg tds after extraction under local anaesthesia of impacted third molars on one side of the mouth. The medication, which was both patient and operator blind, was reversed after extraction of the contralateral third molars 3 weeks later, the patients acting as their own controls in assessing postoperative pain, pain relief, and interincisal opening. Patients receiving diclofenac dispersible recorded significantly lower pain levels; pain relief was significantly greater and the patients' assessment significantly favoured diclofenac dispersible. Interincisal opening throughout the study period was significantly increased in the diclofenac dispersible group. The surgeons' postoperative assessment of extraction sites showed no significant difference between the two treatment groups in rate of healing. Two patients reported side-effects while taking soluble aspirin, and eight while taking diclofenac dispersible, two of whom discontinued treatment. PMID- 8409569 TI - The effect of soft-laser application on postoperative pain and swelling. A double blind, crossover study. AB - The effect of soft-laser application on postoperative pain and swelling was evaluated in a double-blind, crossover study. Twenty-five healthy adults with bilateral identically impacted lower third molars were selected for this study. The teeth were removed in two separate operations. Laser treatment was tested in comparison with placebo laser, with a 40-mW, 830-nm Biophoton laser (Roenvig Dental, Denmark). All surgical procedures and measurements were done by the same surgeon. The following features were statistically analyzed: swelling, trismus, and subjective registration of pain on a visual analog scale. No statistically significant differences were observed in comparison of the experimental side with the placebo side. It may be concluded that soft-laser treatment has no beneficial effect on swelling, trismus, and pain after third molar surgery. PMID- 8409570 TI - Immune response to nickel and some clinical observations after stainless steel miniplate osteosynthesis. AB - Fifteen patients undergoing internal fixation for mandibular fractures were examined for delayed type hypersensitivity (DTH) reaction to nickel before and after implantation of stainless steel miniplates. A lymphocyte transformation test (LTT) was used to measure the immune response before implantation and at removal of plates and screws. The clinical complications during fracture healing were also recorded. The results show that the in vitro LTT may be useful in nickel hypersensitivity testing. A significant increase in lymphocyte transformation was found as a response to nickel stimulation (0.5 microgram Ni/ml) at removal of the implants. We conclude that, despite an increase in LTT after stainless steel implantation, no adverse clinical complications related to DTH could be recorded after short-time observation. PMID- 8409571 TI - International guidelines for specialty training in oral and maxillofacial surgery. PMID- 8409572 TI - Prognostic factors in acute pancreatitis. PMID- 8409573 TI - DNA aneuploidy is an independent factor of poor prognosis in pancreatic and peripancreatic cancer. AB - The purpose of this study was to investigate the clinical significance of DNA ploidy, as assessed by flow cytometry, for pancreatic and peripancreatic cancers. Between 1988 and 1990, we examined fresh/frozen samples from 49 patients who had histologically confirmed adenocarcinomas of the bilio-pancreatic carrefour: They had 23 cancers of the pancreas, 21 of the Vater's papilla, and 5 of the common bile duct. All patients were selected among a cohort of subjects who underwent Endoscopic Retrograde Cholangio Pancreatography (ERCP) and/or surgery. No prognostic impact of age, sex, stage, and surgical treatment on survival was observed by univariate analysis. When the affected organ was considered, a statistically significant difference in survival was observed: At 88 wk, survival was 0% for pancreatic and common bile duct cancer patients, and 18.2% at 175 wk for Vater's papilla cancer patients (p = 0.04). In addition, we found, irrespective of affected organ, that the patients with DNA diploid tumors had a statistically significant survival advantage as compared to those with DNA aneuploidy (p = 0.02). Furthermore, the statistically significant prognostic power of DNA ploidy was confirmed when patients with tumors of the pancreas and those with tumors of the Vater's papilla were separately analyzed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409574 TI - Nuclear DNA content as a prognostic predictor in carcinoma of the pancreas. AB - Eighty-six patients with carcinoma of the pancreas were studied retrospectively. Paraffin-embedded specimens and flow cytometry were used to evaluate the accuracy of the measurement of nuclear DNA as a predictor of the postoperative prognosis. From the series of 86 patients, 72 with a diagnosis of tubular adenocarcinoma (Japanese classification) were selected, and their DNA ploidy pattern and clinical and pathological features were correlated; 52.3% of the 86 patients and 52.8% of the 72 tubular adenocarcinoma patients showed DNA aneuploidy. Histological examinations of the tubular adenocarcinomas showed 42.9% DNA aneuploidy in well differentiated, 56.8% in moderately differentiated, and 71.4% in poorly differentiated types. The DNA ploidy showed a statistically significant positive correlation with the T category. The presence or absence of retroperitoneal invasion was thought to be the most important prognostic factor. Cumulative survival rates showed that the prognosis for patients with retroperitoneal invasion and DNA aneuploidy was significantly worse than for those with DNA diploidy or those without retroperitoneal invasion. PMID- 8409575 TI - Function tests in the diagnosis of chronic pancreatitis. Critical evaluation. AB - Direct pancreatic function tests, such as the secretin-pancreozymin test and the Lundh test, are--by direct measurement of the contents of duodenal secretion following exogenous (hormonal) or endogenous (test meal) stimulation--the best way to assess exocrine pancreatic function. However, these tests are time consuming, invasive, and expensive, and therefore their use is limited to specialized centers only. Several studies have shown that indirect pancreatic function tests are a practicable alternative to direct pancreatic function tests for diagnosing exocrine pancreatic insufficiency. They allow staging of the disease and thereby facilitate comparison of different studies. They are also useful in some cases for determining whether pancreatic enzyme replacement therapy is necessary or not. In contrast to morphological procedures, they involve neither side effects, risks for the patient, nor complications for patients, investigators, and laboratory staff alike. The procedures being standardized, the test results depend to a lesser extent on the experience of the investigator, and more on the selection of and the instructions to the patient, and to some extent, on the severity of the exocrine pancreatic insufficiency. Like all pancreatic function tests, the indirect tests do not give a clue to etiology nor help investigators to differentiate between pancreatic insufficiency owing to chronic pancreatitis and that owing to pancreatic cancer. The role of pancreatic function tests in diagnosing early chronic pancreatitis in comparison to morphological examinations remains to be established. PMID- 8409576 TI - [Determination of neutrophil function in the respiratory infection by chemiluminescence (CL). I: Change in neutrophil's CL by chemotherapy against the bacterial pneumonia]. AB - We measured neutrophil's CL (CL-index) in 12 patients of the bacterial pneumonia three times per each case: before, during and after chemotherapy. Before the initiation of chemotherapy. CL-index in the six patients remained higher than that in healthy controls, while the remaining six showed lower levels of CL-index compared to the controls. In the 11 cases, their CL-indexes fell to levels lower than those obtained before treatment. Additionally, in the 11 cases their CL indexes increased after the termination of chemotherapy. Furthermore, in the nine cases the product of the neutrophil number and CL-index was decreased by chemotherapy, and the decrease in the product correlated with improvement in their clinical conditions. PMID- 8409577 TI - [Disseminated Trichosporon beigelii infection: report of nine cases and review]. AB - Trichosporon beigelii (formerly called Trichosporon cutaneum) is an emerging pathogen of disseminated trichosporonosis in immunocompromised patients. We conducted postmortem microbiological examinations of lung aspirates and blood in the heart. T. beigelii was isolated from the samples of 7 (2.24%) of 313 patients. Five of 7 patients isolated were diagnosed as having trichosporonosis based on histochemical and clinical findings. The other two were considered as colonization. Immunohistochemical study using Cryptococcus neoformans-absorbed antiserum to T. beigelii was carried out on the sections of autopsied tissues with deep mycoses. The sections from seven patients were positively stained, which were formerly diagnosed as candidiasis and/or aspergillosis. Consequently, 9 patients had disseminated trichosporonosis caused by T. beigelii. In Japan, 43 patients including the present 9, 31 males and 12 females, aged 51 (range, 2-84), were reported in the literature. Thirty-seven (86%) patients had hematologic malignancy, and the majority of them revealed profound neutropenia due to cytotoxic chemotherapy. Thirty-eight (88%) patients died despite the anti-fungal chemotherapy including amphotericin B. New strategies for refractory disseminated trichosporonosis in immunocompromised patients is needed. PMID- 8409578 TI - Endometrial bacterial flora detected in patients with uterine endometrial cancer. AB - Certain bacteria produce some carcinogens such as N-nitro compounds, n-butyric acid and n-valeric acid. From this point of view, the examination of intrauterine bacterial flora in patients with uterine endometrial cancer may provide important information. Twenty patients with the diagnosis of uterine endometrial cancer and 20 patients without complications other than myoma uteri were enrolled in the study. Enterobacteriaceae, Streptococcus agalactiae and anaerobic bacteria were mainly detected. The products of these bacteria might be considered to contribute to the initiation of endometrial carcinogenesis. Mixed abnormal flora between aerobic and anaerobic bacteria were detected in all patients with uterine endometrial cancer. It is suggested that uterine endometrial cancer provides favorable conditions for bacterial growth. Mixed abnormal bacterial flora also might influence the onset and growth of uterine endometrial cancer. PMID- 8409579 TI - [Isolation of penicillin-resistant Streptococcus pneumoniae in Saga Medical School Hospital]. AB - A recent nationwide increase in beta-lactams-resistant Streptococcus pneumoniae has attracted a great deal of attention. We studied the drug sensitivity of S. pneumoniae isolated from various clinical specimens in Saga Medical School Hospital between April 1988 and December 1991. To determine the drug sensitivity of the strains, we used a micro-dilution method and determined the MIC. Drug resistance was evaluated using MIC of ampicillin (ABPC) as a reference MIC, and the results were roughly classified into the following three groups: sensitive (< or = 0.1 microgram/ml), moderately resistant (0.2-3.13 micrograms/ml) and highly resistant (> or = 6.25 micrograms/ml). The isolation frequency was calculated on the basis of one strain from one patient. No strain of S. pneumoniae with high resistance against ABPC was found in 1988 (94 strains of S. pneumoniae were isolated) and 1990 (115 strains isolated), but one such strain (0.8%) was found among 129 strains isolated in 1989, and 2 such strains (2.4%) among 84 strains isolated in 1991. Moderately resistant strains were isolated at the frequencies of 12.8%, 15.5%, 22.6%, and 21.4% respectively, in 1988, 1989, 1990, and 1991. A sum of the frequencies of "moderately resistant" and "highly resistant" (2.4%) strains was 23.8% in 1991. The frequency of resistant strains is increasing and the intensity of resistance is also being elevated. PMID- 8409580 TI - [Identification of enterotoxin-producing Clostridium perfringens by the polymerase chain reaction]. AB - Polymerase chain reaction was applied to identify enterotoxin-producing Clostridium perfringens by amplifying a segment of the C. perfringens enterotoxin gene. All of the four enterotoxin-positive reference strains tested were PCR positive while an enterotoxin-negative strain was PCR positive. All 17 clostridial strains (16 species) other than C. perfringens were PCR negative. With clinical strains isolated from various clinical specimens in Japan, Korea, and Thailand, all three enterotoxin-positive isolates were PCR positive and all 82 enterotoxin-negative isolates were PCR negative. PCR results for amplifying a region containing the initiation codon of the C. perfringens enterotoxin gene also demonstrated complete agreement with enterotoxin producibility. These results suggested that the PCR assay is a rapid and simple test for identifying the enterotoxin-producing C. perfringens without using any cultures and spore treatments. PMID- 8409581 TI - [Detection of antibody to varicella zoster virus by immune adherence hemagglutination]. AB - Anti varicella-zoster virus (VZV) antibodies were detected by an immune adherence hemagglutination (IAHA) test, and were compared with the CF test, IFA test and ELISA test, respectively. Type O, Rh-positive RBC for IAHA was obtained from five healthy volunteers. All five RBCs had sufficient sensitivity as the indicator cell. Optimum incubation temperature was 37 degrees C in the serum and in the complement. The complement was obtained from guinea pig sera, and the most suitable concentration was 1;100. The convalescent VZV antibody titers were similar to the values obtained from any of the methods previously mentioned. However, mean titers measured by CF were about two-to fourfold lower than in the values of IAHA. Seroconversion rates of the live VZV vaccine, as detected by CF and IFA were relatively low (CF; 76%, IFA; 56%). In contrast, those obtained from ELISA and IAHA showed perfectly (100%). These results indicate that IAHA has sufficient sensitivity and specificity in the detection of VZV antibodies. In addition, the IAHA test is thought to be a rapid and easy test to perform in ordinary laboratories. PMID- 8409582 TI - [Mechanism of pore formation on erythrocyte membrane by streptolysin-O]. AB - The erythrocyte membrane damaged by streptolysin-O (SLO) was observed in negative staining electron microscopy. It was confirmed that rings took arc (c-ring), sigmoidal (s-ring) or circular (o-ring) structures, and had electron-dense centers of a diameter of 24 nm and 4.9 nm width. We found a crown structure on top of the ring in view of side projection. The ring structure was constructed by three layers of the electron lucent top which was the crown, the second dark layer, and the third, base part which embedded in the erythrocyte membrane, and the heights were 3.2, 1.6, 5.0 nm, respectively. When the ghost membrane of erythrocyte was treated with SLO, the double of the inner and outer layers of a ring were observed by the negative-staining images. The figures of rings taken by under focus showed that one ring might be constituted between the 22 and 24 pair of inner and outer molecules. Totally 44 or 48 toxin molecules might be required for one O-ring. PMID- 8409583 TI - [Cytotoxic effect of ammonia produced by Helicobacter pylori urease on the cultural cells]. AB - We investigated the action of the ammonia produced by Helicobacter pylori urease on the cultured cells. The urease was purified from supernatant fluid of sonicated cell of H. pylori cultured on blood agar for 2 days at 37 degrees C under microaerophilic condition. Purification was carried out by DEAE-Sepharose chromatography, Phenyl-Sepharose chromatography, Sephacryl S-200 SF chromatography and fast protein liquid chromatography on Mono-Q. Vero, HeLa and Intestin 407 cells with or without the addition of 30 mM urea were exposed to the purified urease. Those cells showed cytotoxic effects within 80 minutes after addition of purified urease in the presence of urea. The ammonia production was observed on tissue culture medium within 10 minutes, and the ammonia concentration ranged from 5.56 mg/ml to 7.3 mg/ml and pH in the medium was over pH 9.0. No such effect was observed on the cells exposed to urease without urea. Ammonia water added to Vero cells showed the same cytotoxic effect within 70 minutes on the production of ammonia and raised the pH. However, when the cells were exposed to the ammonia water pre-neutralized to a given pH 7-8 using 1 N HCl cytotoxic effect was not observed. It was concluded that the cytotoxic effect of H. pylori urease was dependent on ammonia generated by hydrolysis of urea. PMID- 8409584 TI - [An infection model which was induced in a carboxymethyl cellulose (CMC) pouch on the back of the rat]. AB - The air-pouch model of inflammation in rats is excellent in that it allows quantitative evaluation of inflammation, and it is used for analysis of inflammatory mediators and as an evaluation system for anti-inflammatory drugs. We investigated the possibility of using this system as an experimental infection system. As a result, inflammation was found to be caused by injection of a constant amount of Staphylococcus aureus solution (10(4)-10(8)). The amount of infiltration and the number of infiltrating cells varied with quantity of bacteria. The infiltrating cells consisted mainly of neutrophils. In this experimental model of infectious disease, the severity of inflammation could be quantitatively evaluated as a function of time in terms of bacterial proliferation and the body's response to bacterial proliferation based on the amount of fluid in the air pouch and the number of infiltrating cells, suggesting that the model is useful. In this experimental system, there were no differences between the number of live bacteria, the number of infiltrating cells or the amount of infiltration when S. aureus Smith strain and clinically-isolated methicillin-resistant S. aureus (MRSA) were used, suggesting that there is no difference between the inflammation-induced activity of MRSA and MSSA. PMID- 8409585 TI - [Serotypes of enteroadherent Escherichia coli exhibiting diffuse pattern of adherence isolated from diarrheal patients and healthy controls in Brazil, Myanmar and Japan]. AB - We tried to detect enteroadherent Escherichia coli exhibiting a diffuse pattern of adherence to HeLa cells from stock strains derived from patients with or without diarrhea in Brazil, Myanmar and Japan (Osaka). Enteroadherent E. coli was found from 16 (23 strains) out of 126 (384 strains) in diarrheal infant cases (12.7%), 26 (29 strains) out of 126 (348 strains) in healthy control cases (20.6%) in Brazil, from 15 (18 strains) out of 221 (542 strains) in diarrheal infant cases (6.8%), 4 (4 strains) out of 87 (212 strains) in healthy control cases (4.6%) in Myanmar. In Japan (Osaka), enteroadherent E. coli was detected from 7 (7 strains) out of 123 (198 strains) in diarrheal cases (5.7%). These results show that enteroadherent E. coli (diffuse) may not be associated with diarrhea. Forty-four (62.9%) out of 70 strains belonged to 19 O serogroups or 21 O:H serotypes. All strains were H antigen serotypable, and 26 strains were found to be nonmotile. The predominant O:H serotypes were O11:H15, O11:H-, O20:H34, O21:H5, O89:H-, O99:H33 and O154:H45. Only one strain belonged to enteropathogenic E. coli serogroup O127. PMID- 8409586 TI - [Development of anti-Treponema pallidum-IgM antibodies detection methods using purified antigen--comparison with conventional methods]. AB - Treponema pallidum subsp. pallidum Nichols (Tp) antigens were purified by centrifugation in the presence of Hypaque (sodium diatrizoate) or MgCl2. When the TPHA (Treponema pallidum hemagglutination) tests with crude and purified antigens were carried out in the sera of patients with untreated primary syphilis, the purification by MgCl2 resulted in enhanced sensitivity to Tp-specific IgM as the same as the purification by Hypaque. In addition, the SPHA (solid phase hemadsorption) test with the Hypaque-purified antigen to anti-Tp-IgM antibodies was carried out using the sera of patients with primary and secondary syphilis. Fifty-nine percent (27/46) of them showed positive for the crude antigen and 100% (46/46) for the purified antigen, indicating that sensitivity to anti Tp-IgM was clearly enhanced with the antigen purification. The captured Tp-IgM-RIA test with the purified antigen was also undertaken in 16 patients with primary syphilis, 31 patients with secondary syphilis and 73 patients with latent syphilis. All the patients in the former two groups were positive, while all of the patients in the latter group were negative. To elucidate the mechanism whereby the purification improves the sensitivity to anti-Tp-IgM antibodies, host-derived antibodies associated with crude and purified Tp cells were measured by a fluorescent antibody technique. As the results, a great amount of rabbit anti-Tp-IgM antibodies were found on the surface of the crude antigen, whereas only a trace amount was detected on the purified antigens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409587 TI - [Anti-H. pylori antibodies of the patients with gastric cancer]. PMID- 8409588 TI - [Efficiency of porin as a component vaccine to Salmonella infection]. PMID- 8409589 TI - [A study on the detection of Helicobacter pylori in children with gastric and duodenal diseases]. AB - Infection of Hellcobacter pylori (H. pylori) in the gastric mucosa is said to participate in the pathogenesis of gastric and duodenal ulcers as well as gastritis. The detection rate of H. pylori in tissue specimens endoscopically taken from adult patients with these diseases was 75.2% (408/542). In affected children (2-15 yrs), on the other hand, 27 biopsies were performed in 22 cases but the initial positive rats was as low as 45.5%. According to the diagnosis, it was 75% for acute gastric mucosal lesion (AGML), 50% for duodenal ulcer, 33% for duodenitis, and 38% in seemingly normal children. When changes in the detection rate of H. pylori was examined after administration of H2 blocking antiulcer agents, it was found to very case by case, e.g. changed in individual disease stages or completely or reoccurrence disappeared. PMID- 8409590 TI - [Immunogenicity of Chinese hamster ovary cell derived HB vaccine--a comparative study with yeast derived HB vaccine]. AB - To evaluate the immunogenicity of the Chinese hamster ovary cell derived HB vaccine (CHB), CHB and yeast derived HB vaccine (YHB) were given to 110 medical students divided into four groups. CHB was given to group A subcutaneously (0.5 ml, 3 times) and to group B intradermally (0.125 ml, 3 times). YHB was given to group C (Y company) and to group D (Z company) subcutaneously (0.5 ml, 3 times). The anti-HBs positive rate at 7 months was 85.7% in group A, 95.8% in group B, and 100% in groups C and D. The percentage responding with high anti-HBs concentration (> or = 100 mIU/ml by RIA) at 7 months was 67.9% in group A, 79.2% in group B, 58.3% in group C and 63.3% in group D. Thus, although the anti-HBs positive rate was rather low when CHB was given subcutaneously, it was almost the same as YHB when given intradermally. The percentage of high responders to CHB given both subcutaneously and intradermally was high compared to YHB. These data suggest that CHB has almost the same immunogenicity to the conventional YHB. PMID- 8409591 TI - [Current status of Pasteurella multocida infection in Japan]. AB - Recently, the case reports of Pasteurella multocida infection has been increasing in Japan. In 1989, the Japanese Government, Veterinary Sanitation Division, Ministry of Health and Welfare officially communicated this infection as a zoonosis to related institutions. The current status of Pasteurella multocida infection is not well known in Japan. Because of this, a nation wide questionnaire survey on Pasteurella multocida was conducted to clarify the status. A questionnaire was sent to 380 laboratories of the hospitals, and 258 (67.9%) replied. An infectious disease caused by Pasteurella multocida was found in Japan in 369 cases in 115 (44.6%) of 258 hospitals, or an average of 3.2 cases per hospital. The 369 cases were broken down into 123 males (from 1 month old to 87 years old), 118 females (from 7 months old to 88 years old), and 128 patients whose sex was unknown. The incidence of the infections tends to increase year by year. This incidence is higher than our expectation. It is considered that the contact with pets will in increase the infection with this agents. The organism was isolated in as many as nineteen different body specimens, including the appendix and urine, which in Japan has not been reported as organs harboring this organism. Some of the nineteen cases were severely infected. This organism was isolated most often from the sputum (48.5%). Pus was the next most common site (27.1%). This order was reversed in the U.K. and the U.S.. Possible explanations for the reversal are given below.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409592 TI - [Prevalence of bacterial pathogens and antimicrobial susceptibility: a multicenter study in Akita prefecture]. AB - We report the prevalence of bacterial pathogens isolated from various clinical specimens in 17 medical centers in Akita during the period of December 1990 to February 1991, and their antimicrobial susceptibility. The results were as follows: 1. The total number of isolates was 8,929 and common bacteria were Staphylococcus aureus (18.7%), Pseudomonas aeruginosa (11.8%), Escherichia coli (10.1%), coagulase negative staphylococcus (6.7%) and Enterococcus faecalis (5.0%). This is similar to our previous result which was reported in the same season in 1987. 2. S. aureus was a common isolate from sputum, throat swab, pus, and blood. 53% of isolated S. aureus were methicillin-resistant. Arbekacin (ABK) had a high degree of activity (99%) against methicillin-resistant S. aureus (MRSA). Minocycline (MINO) retained activity against 70% of MRSA strains, and quinolones only 20%. 3. P. aeruginosa and E. coli were frequently found in urine from the patients in the small hospitals. More than 80% of the P. aeruginosa strains were sensitive to imipenem (IPM), cefsulodin (CFS) and amikacin (AMK). The susceptibility of P. aeruginosa to ceftazidime (CAZ) and piperacillin (PIPC) had been decreasing for the last 3 years. 4. CNS were frequently found in blood, pus, and pleural effusion from the patients in the large hospitals. The isolates were resistant to most antibiotics. 5. Enterococcus was common isolates from urine and ascites, the susceptibility to cephem was low. 6. We conclude that attention should be specially paid to S. aureus, P. aeruginosa CNS and E. Faecalis. PMID- 8409593 TI - [Antibacterial activity of cefdinir and omeprazole against Helicobacter pylori and their inhibition on H. pylori-producing urease]. AB - Since Helicobacter pylori is isolated very frequently from gastric ulcer specimens, the combination therapy of antimicrobial agent and proton pump inhibitor has recently been used. A study was made on whether cefdinir (CFDN), amoxicillin (AMPC), metronidazole (MNZ), omeprazole (OPZ), and omeprazole-M (OPZ M) have antimicrobial activity against H. pylori and whether they can inhibit H. pylori-producing urease. 1) CFDN, AMPC and MNZ showed a potent antimicrobial activity against H. pylori, and especially, AMPC showed a marked bactericidal activity in a short time. 2) OPZ is reported to be converted to OPZ-M, and active form, in the body. OPZ and OPZ-M showed a moderate antimicrobial activity against H. pylori, and scarcely any bactericidal activity. 3) CFDN and OPZ or AMPC and OPZ in combination did not show any synergistic effect on the antimicrobial activity, but MNZ and OPZ in combination showed additive effect on the antimicrobial activity against H. pylori. 4) OPZ and OPZ-M inhibited H. pylori producing urease and the inhibitory effect of OPZ-M was more stronger than that of OPZ. CFDN, AMPC and MNZ did not show any inhibitory effect on H. pylori producing urease at 10 micrograms/ml. From these data, antimicrobial agents and proton pump inhibitors in combination are expected to exert the in vivo synergistic effect since these drugs eradicate H. pylori and inhibit H. pylori producing urease. PMID- 8409594 TI - [Two cases of methicillin-resistant Staphylococcus aureus infection]. AB - We have experienced 2 cases of MRSA infection. Case 1: A 16 month-old girl, whose underlying disease was VAHS, had chronic sinusitis. MRSA was isolated from the blood and rhinorrhea. Her sinusitis was very intractable and it was difficult to eradicate MRSA from the nare. Case 2: A 6-months-old girl was admitted to our hospital with urinary tract infection. She had an ectopic uretelocele. Partial nephrectomy and uretectomy was performed. Retroperitoneal and intraperitoneal abscess had recurred three times during 17 months after the first operation. This abscess was slowly progressive and MRSA was very difficult to eradicate. These 2 cases showed characteristics of MRSA infection. PMID- 8409595 TI - [Successful 24-hour pulmonary preservation in the use of hyperbaric oxygen]. AB - 24-hour lung preservation with modified Euro-Collins (E-C) solution under hyperbaric oxygen (OHP) was studied in the canine single lung transplantation model. A prostaglandin I2 (PGI2) was administered before cardiac arrest by St. Thomas solution. Lungs were flushed with modified E-C solution, then heart and lungs were excised en-bloc and immersed in 4 degrees C modified E-C solution for 24 hours in control group (N = 5). In OHP group (N = 5), heart and lung block were preserved similarly in the control group but placed in hyperbaric chamber (mixed gas 95% O2, 5% CO2) under 2 atmospheres absolute pressure. In the hyperbaric chamber, the lungs were expanded continuously by pressure difference between the inside and the outside of the lung. The left lung was transplanted into recipient dog and reperfused for 3 hours. Pulmonary function was assessed serially by measuring oxygen and carbon dioxide tensions in arterial blood and pulmonary vascular resistance by clamping the right pulmonary artery for 5 minutes. ATP before and after preservation was measured. In the control group, PaO2, after 24 hours preservation followed by reperfusion, was 177 +/- 88 mmHg, but decreased to 75 +/- 34 mmHg at 3 hours of reperfusion and bloody foamy sputum appeared in 4 dogs. Alternatively, PaO2 was 256 +/- 58 mmHg at 3 hours of reperfusion and foamy sputum did not appear in OHP group. There was a significant difference between the 2 groups (p < 0.05). PaCO2 at 3 hours of reperfusion was 41 +/- 3 mmHg in the control group, and 31 +/- 2 mmHg in the OHP group (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409596 TI - [Results of surgery for pT4 lung cancer]. AB - During 1977 and 1991, 54 patients with lung cancer underwent surgery at Niigata University Hospital and were diagnosed with pathological T4. The survival rate of these pT4 patients was 32.5% at 3 years and 24.4% at 5 years. There was no significant difference between the survival rates of those with squamous cell carcinoma and those with adenocarcinoma. 5-year survival rate of 20 patients with N0 disease and 23 patients with N2 disease was 43.0% and 13.0%, respectively (p < 0.05). 5-year survival rate of 21 patients with organ invasion alone, 21 with dissemination or malignant effusion alone, and 12 with organ invasion plus dissemination or malignant effusion was 22.5%, 28.6% and 0%, respectively; there was no significant differences between these rates. Seven patients survived over 5 years; however, there were no obvious common factors. An aggressive surgical approach is indicated for T4 lung cancer, not only in patients with N0 disease but also in those with organ invasion alone or small amounts of malignant effusion alone. PMID- 8409597 TI - [The long-term patency rate of saphenous vein grafts and vein graft disease in Japanese patients]. AB - While annual attrition and high break-down rate of saphenous vein graft (SVG) used for CABG has widely noted in Western countries, no sizable studies have yet available in Japan. We studied 142 SVGs of 77 pts, which we divided into two groups; 80 SVGs of 44 pts in mid-term period (5 to 8 years after surgery) and 62 SVGs of 33 pts in long-term period (9 to 17 years after surgery). The patency rate of SVGs was 69% in mid-term and 77% in long-term. Whereas these patency rates at each periods were superior to those reported from USA and European countries, a quarter of SVGs in mid-term period and a half in long-term period had significant stenotic changes (over 50%). Of patent SVGs, diseased SVGs reached 36% in mid-term period and 73% in long-term period. In conclusion, although the patency rate of SVGs in Japanese patients was higher than that of the Western countries, vein graft disease apparently occurred in a large proportion of patent SVGs. PMID- 8409598 TI - [Correlation of myocardial temperature, intramyocardial pH, and myocardial electrical activity during hyperkalemic hypothermic cardioplegic arrest to functional recovery after reperfusion]. AB - We estimated the correlation of myocardial temperature, intramyocardial pH, and myocardial electrical activity (MEA) during hyperkalemic hypothermic cardioplegic arrest to functional recovery after reperfusion to assess those parameters as intra-operative on-line assessment of myocardial preservation during cardiac operation. Twenty dogs underwent cardiopulmonary bypass at 30 degrees C. Cardiac function was evaluated before and after two hours ascending aortic cross clamping. The dogs were divided into four groups according to cardiac preservation methods employed. Group I (n = 6): 4 degrees C hyperkalemic crystalloid cardioplegic solution was delivered antegradely through the aorta every 30 minutes without topical cooling. Group II (n = 4): total volume of cardioplegic solution same as Group I was delivered only once without topical cooling. Group III (n = 5): cardioplegic solution same as Group I with lidocaine (100 mg/l) was delivered every 30 minutes without topical cooling. Group IV (n = 5): cardioplegic solution same as Group I was delivered every 30 minutes with topical cooling. Hearts at less than 8 degrees C during aortic cross-clamping (Group I) revealed better functional recovery (72 +/- 21%) than those over 15 degrees C (Group II-IV, 28 +/- 30%). There was no significant correlation between intramyocardial pH changes during aortic cross-clamping and functional recovery after reperfusion (R = -.2257). During aortic cross-clamping, the occurrence of MEA which was obtained directly from the electrode inserted into left ventricular anterior free wall (direct MEA) was observed in all cases. However the myocardial activities which were identified by direct MEA were not necessarily observed visually or by routine electrocardiogram.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409599 TI - [Long-term results of closed mitral commissurotomy--comparative study of closed mitral commissurotomy (CMC), open mitral commissurotomy (OMC) and mitral valve replacement (MVR)]. AB - As the technique of open heart surgery has improved, CMC has been abandoned in favor of OMC and MVR. We evaluated and compared the results of CMC, OMC and MVR. METHOD. Between 1965 and 1978, 141 patients with mitral stenosis (MS) underwent CMC, and late follow-up obtained in 117 (83%) of them (CMC group). Between 1980 and 1989, 72 patients and 37 patients underwent OMC (OMC group) and MVR (MVR group), respectively. Cumulative follow-up periods were 1982, 632 and 200 patient years in the CMC, OMC and MVR groups, respectively. RESULTS. (1) Survival rate; In the CMC group there were 2 operative deaths due to severe mitral regurgitation (MR). There were 17 late deaths, due to reoperations in 4 patients, cerebral infarction in 4 patients, congestive heart failure in 3 patients, myocardial infarction in 2 patients and unknown causes in 4 patients. The survival rate was 95%, 91% and 86% at 5, 10 and 15 years, respectively, in the CMC group. In the OMC and MVR groups there was no death. (2) The event free rate was 89%, 79% and 58% at 5, 10 and 15 years, respectively, in the CMC group, 97% and 97% at 5 and 10 years in the OMC group, and 95% and 90% at 4 and 5 years in the MVR group. (3) Reoperations; In the CMC group of 40 patients (34%) required reoperations in an average of 10.4 years after the initial operation, due to re-MS in 22 patients, MR in 10 patients and MRS in the 8 patients. Reoperative findings consisted of clefts in the mitral leaflets in 7 patients. There were pulmonary hypertension in 15 patients and tricuspid regurgitation in 22 patients. Fourteen patients underwent tricuspid anuloplasty and one patient underwent a tricuspid valve replacement. In the OMC group one patient required a reoperation due to MR; in the MVR group one patient required a reoperation due to a thrombosed valve. CONCLUSION. In the CMC group the survival rate and the event free rate were lower, and the rate of reoperation was higher than in the other two groups. PMID- 8409600 TI - [A clinical and experimental study of the antithrombotic percutaneous cardiopulmonary support using the new antithrombotic coating material]. AB - The antithrombogenicity of the new antithrombogenic coating material "Fluorine acryl-styrene-urethane-silicone (FASUS) graft-block copolymer" was evaluated by using arerio-arterial (A-A) shunt method in a rabbit and heparinless veno arterial bypass (VAB) model in a dog. In ex vivo A-A shunt model, FASUS coated group (n = 4) showed significantly longer occlusion times compared with CONTROL group (n = 4) as follows (mean +/- SD): FASUS 109.5 +/- 34.7; CONTROL 3.0 +/- 0.8 minutes. In heparinless VAB model, all tubing materials from the CONTROL group (n = 6) showed major clots, by contrast, no clots were observed in any part of the entire circuits of the FASUS coated group (n = 6) (p < 0.005). The scanning electron microscopic examination revealed thick layers of platelets aggregate and fibrin sheets have firmly adhered to the uncoated surfaces (p < 0.05). In the FASUS coated group, morphologic platelet changes were slight. And we used the cannulas coated by FASUS copolymer in percutaneous cardiopulmonary support (PCPS) with low dose systemic heparinization equipped by heparin coated oxygenator, centrifugal pump and tubing materials. And activated coagulation time was maintained about 150 seconds with or without minimal systemically administered heparin. Consequently, the result was significantly improved (weaning rate 72.2%, long survival rate 33.3%), and hemorrhagic or thromboembolic complications were not observed. These data suggest that FASUS copolymer coating is effective for preventing thrombus formation, and PCPS with low dose systemic heparinization is useful to control postoperative bleeding. PMID- 8409601 TI - [Preoperative diagnosis of the thoracic aortic aneurysm by three-dimensional CT angiography]. AB - Serial eight patients with thoracic aortic aneurysms were evaluated by a newly developed three-dimensional CT angiography (3D-CT) from December 1992 to January 1993. The patients include 3 aortic dissections, 3 aortic arch aneurysms, one descending aortic aneurysm and one thoracoabdominal aortic aneurysm. The surgical treatment was performed after the evaluation of 3D-CT, and the operative findings were compared to the three-dimensional images reconstructed by 3D-CT in all patients. Three-dimensional displays were achieved using the unique method of data collection of the helical (spiral) scanner with continuous tube rotation and continuous table feed. A intravenous contrast material was used to image the thoracic aorta and major aortic branches with the single-breath-hold technique. Two and three-dimensional images reconstructed by 3D-CT were displayed within 10 20 minutes after the scanning. These three-dimensional images of the aortic lesions could be displayed in any angle we chose. Three-dimensional structures of the thoracic aorta and major aortic branches were clearly visualized and easily recognized by 3D-CT. These images were similar to the intraoperative findings and were quite useful to determine the operative procedure. The successful repair of thoracic aortic aneurysm was achieved in all cases. 3D-CT is a new and attractive modality to assess the vascular system. Although our experience is limited, 3D-CT may be a useful and powerful diagnostic method for the surgical treatment of thoracic aortic aneurysm. PMID- 8409602 TI - [Myocardial protection with pulsatiled retrograde coronary sinus perfusion--using Emax for left ventricular contractility]. AB - Recently retrograde coronary sinus perfusion technique has been frequently used in patients with severe coronary artery disease. However many untoward effects, such as tissue edema and hemorrhage, have also noticed. To evaluate the efficacy of retrograde cardioplegia, 24 mongrel pentobarbital anesthetized dogs were studied. To create hypoperfused area, distal portion of the left anterior descending coronary artery (LAD) was occluded. After cardioplegic arrest under cardiopulmonary bypass, dogs were assigned following 3 experimental groups (8 dogs each). Group I; Glucose-Insulin-Potassium (GIK) solution (K: 20 mEq/l, 20 ml/kg) was given antegradely into the aorta. Group II; GIK was given retrogradely through the coronary sinus. Group III; GIK was given retrogradely with pulsatile device (synchronized retroperfusion pump system). After 30 minutes, same amount of GIK was given again. Then LAD occlusion was released. Sixty minutes after onset of arrest, the aorta was declamped and cardiopulmonary bypass was stopped. The left ventricular contractility (Emax) measured with conductance catheter at the end of experiment was significantly (p < 0.05) better in groups II (13.1 +/- 2.6, mean +/- SD) and III (13.1 +/- 2.9) than in group I (9.6 +/- 2.7). The left ventricular wall-motion measured with ultrasound crystals in hypoperfused area compared to before cardiopulmonary bypass was also significantly better in groups II (76.2 +/- 17.2%) and III (87.9 +/- 16.9%). Regional myocardial temperature suggested that more rapid and homogeneous cooling including right ventricle was achieved in group III than in groups I and II. Retrograde perfusion is more effective method in the setting of coronary stenosis compared to ordinal antegrade technique.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409603 TI - [Efficacy of myocardial preservation using HTK solution in continuous 120 min cross-clamping method--a comparative study with GIK method]. AB - The aim of this study is to investigate the efficacy of HTK solution in the 120 minutes cross-clamping method in comparison with conventional intermittent cardioplegia using GIK solution. Fifty-four open heart surgery were performed with cardioplegic solution using either HTK solution (HTK) or GIK solution (GIK). In the HTK, HTK (3L) was infused for the initial dose and 1L was added every 60 min after 120 min of cross-clamping. In GIK, 1L of GIK solution was intermittently infused initially and then every 30 min together with continuous cold blood perfusion. The effect of two cardioplegic solution was evaluated by postoperative cardiac function (CI, %SF), released enzymes (CPK), histology and dosage of catecholamine. Postoperative CI was 3.67 +/- 0.76 in HTK, and 4.34 +/- 1.04 in GIK (NS). % SF was 26.0 +/- 5.26 in HTK and 25.6 +/- 0.76 in HTK, and 4.34 +/- 1.04 in GIK (NS). %SF was 26.0 +/- 5.26 in HTK and 25.6 +/- 9.2 in GIK (NS). The CK-MB (IU/dl) level after reperfusion was significantly decreased in HTK at 60 and 180 min after reperfusion. Histology at 60 min of ischemia revealed a significant increase of edema of mitochondria in GIK. Postoperative catecholamine dose was 2.65 +/- 1.3 in HTK and 10.3 +/- 3.4 in GIK (p < -0.01). PH of myocardium was well maintained around 7.4 during cross-clamping in HTK, however, it was decreased in GIK. In conclusion, The HTK method offers a reliable cardiac protection due to effective buffering using Histidine in comparison with GIK. PMID- 8409604 TI - [Intracavitary suction for treating giant bulla]. AB - Examination was made of 9 consecutive patients who underwent intracavitary suction for giant bulla. A second operation was required for one patient who had multiple giant bullae. In the other 8 patients, improvement of pulmonary function and symptoms was obtained immediately following the operation. This was particularly more apparent in patients with poor pulmonary function. The following results were obtained. Functional recovery was evident from the early postoperative phase. A one-stage operation was possible. Reduction in postoperative drainage time was realized by bronchial occlusion. The present operation is particularly applicable to compromised patients. Computed tomography scanning is essential for evaluation of the drainage site. For giant bulla with some septation and multivesicular, the present mode of treatment would not be indicated. PMID- 8409605 TI - [Imaged thoracoscopic surgery for bronchogenic cyst in posterior mediastinum--a case report]. AB - A 44-year-old female with bronchogenic cyst in the posterior mediastinum was treated successfully by employing imaged thoracoscopic surgical methods in the same way as laparoscopic cholecystectomy. The patient was placed in the left lateral decubitus position. A first 3.5 cm skin incision was made in the seventh intercostal space on the mid-axillary line. A 10 mm, 0-degree Olympus wide angle telescope was inserted for observation into the thoracic cavity. And three more small skin incisions were done. A grasping forceps, a padron retractor and an electrode with J-hook tip were introduced through each incision. The cyst was completely dissected almost with the J-hook electrode. She was discharged on the 10th-postoperative day without complication and with minimal pain. We suggest that an indication of the imaged endoscopic surgery could be established for some benign mediastinal tumors. PMID- 8409606 TI - [Two-staged repeat myocardial revascularization through the sternal re-entry and the left thoracotomy with coronary anastomosis under the beating heart]. AB - A case report of serial repeat myocardial revascularization under the beating heart. A 56-year-old, hypertensive and hyperlipidemic male suffered from unstable angina who had undergone primary CABG with saphenous veins 10 years ago. The vein graft to the left anterior descending coronary artery (LAD) was solely patent but had a severely stenotic lesion. The stenotic LAD vein graft fed all coronary circulation. The second operation was approached through the sternal reentry but E-PTFE sheat for pericardial closure at the primary operation tightly adhered to an anterior aspect of the heart with abscess formation. So only gastroepiploic artery (GEA) could be anastomosed to the right coronary artery (RCA) under the beating heart without cardiopulmonary pump (CPB). While the postoperative angiogram showed GEA graft was patent, unstable angina recurred. Then the third operation was performed 2 weeks later through the left thoracotomy under the beating heart with CPB. A new saphenous vein was anastomosed from the descending aorta to LAD. He recovered well without any major complications. Postoperative angiogram showed two new grafts widely patent and he discharged with freedom from angina. PMID- 8409607 TI - [Surgical treatment of the primary cardiac rhabdomyoma in the neonate--a case report]. AB - A successful surgical treatment of intracardiac rhabdomyoma in a 12-day-old female is reported. The neonate was first noted to have a cardiac murmur and she was diagnosed cardiac tumor by two dimensional echocardiography. It showed multiple cardiac tumors which occupied the right ventricular outflow tract and interventricular septum. Under the routine cardiopulmonary bypass the tumor of the right ventricular outflow tract was removed. Histological examination revealed a rhabdomyoma. Follow-up evaluation 7 years after operation showed normal growth and development of the patient. A repeated two dimensional echocardiogram revealed no sign of recurrence and the left ventricular cavity was not obstructed by the residual tumor in the interventricular septum. We reviewed 10 cases treated surgically in infancy including three neonate in Japan. PMID- 8409608 TI - [An innovative thoracoscopic surgery for patient ductus arteriosus--a Japanese first case report]. AB - A five-year-old girl with patent ductus arteriosus (PDA) was successfully treated by thoracoscopic surgery, which was the first successful case in Japan. The operation was carried out under general anesthesia with usual endotracheal intubation. Short trocars were inserted through the left intercostal spaces to introduce a flexible video thoracoscope and adequate surgical instruments. After the ductus was carefully dissected and exposed, two titanium clips which were 11 mm in length were applied to interrupt the ductus completely. The continuous heart murmur of PDA was confirmed to disappear by an esophageal stethoscope. Postoperative course was uneventful and the patient was discharged on 6 postoperative day. No residual PDA shunt was revealed by doppler echocardiogram 8 months after surgery. The advantages of thoracoscopic surgery for PDA are: less postoperative pain and discomfort, early recovery and short hospital stay, and cosmetic preservation. Availability of smaller sized surgical instruments should allow this method indicated for smaller children or newborns with PDA. PMID- 8409609 TI - [Surgical treatment of coronary artery-pulmonary artery fistula]. AB - We experienced 4 cases of left coronary artery-pulmonary artery fistula. Two cases had small fistulas associated with atherosclerotic coronary lesions, and the other 2 had large fistulas with aneurysmal enlargement. In the former 2 cases, ligation of the fistulas and closure of the opening of fistula into the pulmonary artery through pulmonary arteriotomy were performed together with coronary artery bypass grafting and left ventricular aneurysmectomy. In one of the latter 2 cases, the fistula arising from the anterior descending branch was ligated and the opening of fistula draining into the pulmonary artery was closed through pulmonary arteriotomy. In another case, both openings of the fistula into the anterior descending branch and the pulmonary artery were closed from inside through incision of the dilated fistula. In all 4 cases, operations were performed using cardiopulmonary bypass and retrograde coronary perfusion, which could afford good heart protection even in cases with coronary lesions and coronary steal phenomenon. All cases went an uneventful postoperative course. Postoperative angiograms showed disappearance of the fistulas in 3 cases. In one case, however, residual fistula was found because a fine fistula might be overlooked. In such a case with complicated fistulas with aneurysmal enlargement, fistulas should be examined carefully through incision of the enlarged anomalous vessels. In this paper, diagnosis, operative indication and treatment for coronary artery-pulmonary artery fistula were discussed. PMID- 8409610 TI - [Chondrosarcoma of the trachea--report of a case]. AB - A case of chondrosarcoma of the trachea in a 64-year-old man was reported. He was pointed out of benign tracheal tumor in September 1988. He was transferred to our hospital in September 1991 with respiratory distress attributable to asthma-like attack after bronchoscopic examination. Under right posterolateral thoracotomy, 1.7 cm length (3 rings) of the mediastinal trachea was resected and the trachea was reconstructed by end-to-end anastomosis with 3-0 Maxon interrupted sutures. The pathological diagnosis was a moderately well differentiated chondrosarcoma (grade II) of the trachea. Postoperative clinical course was uneventful without serious anastomotic complications. No local recurrence was seen 1 year after operation. It is necessary to follow meticulously the patients with chondrosarcoma of the trachea after surgery because of its low grade malignancy. PMID- 8409611 TI - [A successful surgical treatment of traumatic dissecting aneurysm]. AB - Dissecting thoracic aortic aneurysm caused by blunt trauma was reported. A 64 year-old woman involved in automobile accident was transported to our hospital. Chest X-ray showed multiple rib fractures and enlargement of mediastinal shadow. Emergent CT scan and angiography (DSA) revealed aortic dissection (DeBakey IIIb). She suffered from acute renal and respiratory failure soon after admission, but underwent operation 6 months after the trauma because of enlargement of false channel. Graft replacement was performed and postoperative course was uneventful. Histopathologic findings showed dissection of adventitia of the aorta. PMID- 8409612 TI - [A case of successfully operated hypertrophic obstructive cardiomyopathy in a child]. AB - A case of surgical treatment of hypertrophic obstructive cardiomyopathy (HOCM) in a 5-year-old boy is reported. This is the youngest case in Japan. He had been diagnosed as HOCM and treated conservatively with beta-blocker and Ca-antagonist, but his complaints of dyspnea and chest oppression during efforts had increased. Cardiac catheterization showed a peak systolic pressure gradient of 110 mmHg between the left ventricle and systemic artery, and echocardiogram disclosed a systolic anterior motion (SAM) of the mitral valve and mitral regurgitation (MR). At operation transaortic septal myectomy and myotomy were performed according to preoperative findings by echocardiogram and MRI, and a marked reduction in pressure gradient and a decrease in SAM and MR were obtained. Thus, a satisfactory result can be obtained by myectomy and myotomy without mitral valve replacement in a pediatric patient with HOCM, based on the preoperative image analysis. PMID- 8409613 TI - [The use of the reverse latissimus dorsi muscle flap in the management of bronchopleural fistula]. AB - The management of a postoperative bronchopleural fistula remains a major challenge for the thoracic surgeon. We present a series of three patients who underwent closure of the fistulas with the distal flap of the divided latissimus dorsi muscle (the reverse latissimus dorsi muscle flap) and a limited thoracoplasty. The primary resections, which were performed through a standard posterolateral thoracotomy, were right lower lobectomy in two patients and right middle and lower bilobectomy in the other. The initial myoplasty produced prompt fistula closure in 2 patients and failed in the other patient because of a large empyema space. The reverse latissimus dorsi muscle flap could be used for the management of bronchopleural fistula in selected patients who underwent lower lobectomy or middle and lower bilobectomy. PMID- 8409614 TI - [An hourglass type intrathoracic lipoma--a case report of surgical treatment]. AB - We report a rare case of hourglass type intrathoracic lipoma. Until now, there are 114 reported cases on intrathoracic lipoma in Japanese literature, among which 22 are hourglass type. The patient was a 43-year-old woman, who was found to have a pleural tumor on her chest X-ray examination for healthy check-up. Computed tomography showed that it had intra- and extrathoracic portion with almost the same density as the subcutaneous fat tissue. The tumor was excised along with pleura, periostia and intercostal muscle approximately 1 cm apart from it. It was dumbbell-shaped and weighed 30 grams. Microscopically it was found to be intramuscular lipoma. According to Fletcher, intramuscular lipoma was classified into two groups of infiltrative and non-infiltrative type. In their cases, infiltrative intramuscular lipomas recurred up to 19%, mainly secondary to incorrect resection. Our case corresponds to infiltrative type. We consider that intrathoracic lipoma, though benign, have to be resected radically with utmost care. PMID- 8409615 TI - [Leiomyosarcoma of the chest wall]. AB - An unusual case of a primary leiomyosarcoma of the chest wall is presented. The patient was a 41-year-old man and had no symptoms. The chest x-ray showed a tumor with mediastinal involvement. A percutaneous needle biopsy indicated that it was histologically a leiomyosarcoma. Wide en bloc excision of the chest wall was done, including much of the third, fourth and fifth ribs and a piece of lung. The defect was closed with GORE-TEX soft tissue patch. There were three courses of chemotherapy added, but the patient had a local recurrence, which was resected again, two years after the first surgery. It may be thought that wide en bloc excision and closely follow-up are important in the management of a leiomyosarcoma of the chest wall because of its high recurrence rate. PMID- 8409616 TI - [A case report: mitral valve replacement for the patient with essential thrombocythemia]. AB - A 62-year-old man with heart murmur and recent cerebral thrombosis was admitted to our hospital. Examination of his heart revealed mitral stenosis and hematological study confirmed the diagnosis of essential thrombocytosis (ET). After 1100 ml pre-operative phlebotomy, mitral valve replacement with a 29 mm Bjork-Shiley prosthesis was performed utilizing routine cardio-pulmonary bypass using only his prestored blood and moderate hypothermia. We had no difficulty in hemostasis at the operation. Acute gastric bleeding began suddenly as much as 1500 ml on the 4th postoperative day. As medical care was in vain, emergent distal gastrectomy was performed. As we used urokinase for fear of thrombosed valve formation, residual gastric bleeding was continued on. Then, we began to use Gabexate-Mesilate (FOY) instead of urokinase. Retrospectively, Gabexate Mesilate was very useful drug for preventing the left atrium and prosthesis from thrombo-formation until the start of oral intake of Warfarin and Busulfan. To our knowledge, this would be the first case report of open-heart valve replacement surgery in a ET patient in Japan. PMID- 8409617 TI - [Case report of MRSA sepsis required two valve replacement twice a year--trying case with hyperthermal extracorporeal circulation]. AB - We report a case in conjunction with MRSA sepsis, who needed re-mitral valve replacement (re-MVR) and re-tricuspid valve imposition (re-TVI), and who was successfully treated with a number of antibiotics in conjunction with hyperthermal extracorporeal circulation. Initially, we performed MVR and tricuspid valve superimposition on a 62-year-old woman lationing under the MRSA sepsis condition to control against heart failure. However, she developed a fever following the first operation, and MRSA was detected from her blood cultivation. She thus underwent treatment employing many kinds of antibiotics. A thickened C.E. valve at the tricuspid valve cardiac echogram suggested PVE, we performed a second operation of re-MVR and re-TVI about a year after the first operation. We used a tangl of antibiotics during the operation, adding Vancomycin into the extracorporeal circulation, and utilized hyperthermal extracorporeal circulation. This patient's postoperative course was uneventful, with no recurrence arising at 23 months after the second operation. PMID- 8409618 TI - [A case of thymoma showing extensive necrosis]. AB - A 44-year-old man was admitted with fever elevation and dyspnea. Chest X-ray and CT examination indicated a 6 x 4 cm anterior mediastinal tumor, pleural effusion and pericardial effusion. Blood examination showed no inflammation findings. Gallium scintigraphy indicated no uptake at the tumor. He underwent resection of the tumor. Postoperative pathohistological examination showed most part of the tumor fell into necrosis. And remained part of the tumor showed a figure of predominantly mixed type thymoma. Thymoma showing extensive necrosis is rare. PMID- 8409619 TI - [Mitral valve re-replacement for a patient with multiple myeloma]. AB - Mitral valve re-replacement was successfully undertaken in a 70-year-old female patient with primary tissue failure of mitral bioprosthesis and multiple myeloma (MM) through right lateral thoracotomy with minimum dissection. Considering the post-operative anti-coagulant therapy, bioprosthesis was chosen in this complicated case. Patients with MM usually show a bleeding tendency associated with abnormally elevated IgG which covers the surface of coagulation factors. Safety range of serum IgG concentration in the patients with MM who require open heart surgery, have not defined yet. Excessive serum IgG leads the coagulation disorder and organ perfusion problem because of increase of blood viscosity. Those problems become serious when the serum IgG concentration exceed 2,500 mg/dl. (1) Only if serum IgG level below 2,5000 mg/dl, valve replacement will be able to perform safely with minimum dissection through the thoracotomy for patients with MM. (2) Bioprosthesis might be optionally chosen in such case (3). In case of serum IgG level over 2,500 mg/dl, preoperative plasma ex-change should be recommended. PMID- 8409620 TI - [A case of empyema with bronchopleural fistula associated with diabetes and renal failure treated utilizing omental and muscle flap]. AB - The patient was a 75-year-old male who had a right empyema with bronchopleural fistula, which had developed after right upper lobectomy for pulmonary tuberculosis performed 18 years previously. He also had diabetes and renal dysfunction. The fistula failed to close 5 months after open drainage thoracotomy. Accordingly, closure was performed by the intrathoracic transposition of an omental and muscle flaps obtained from the right latissimus dorsi, pectoralis major, and pectoralis minor. At 70 days after the closure, the fistula reopened due to his poor nutritional state, but it closed again spontaneously following intravenous hyperalimentation. In those patients who have empyema with bronchopleural fistula associated with diabetes or renal dysfunction, long-term postoperative care, especially nutritional management, is necessary because of their poor general health condition. PMID- 8409621 TI - [A report of successful surgical management of ASD, VSD and PDA associated with partial DiGeorge syndrome in infant]. AB - We report an infantile male case of ASD, VSD and PDA associated with DiGeorge syndrome. The patient who was 58-day old presented the partial deficiency of cell mediated immunity and normocalcemia before the admission to our hospital. We made semi-emergent operation because of increasing of lung congestion caused by repetitional infection of upper respiratory tract. Direct closure of the ASD, patch closure of the VSD and ligation of the PDA were performed successfully. He was suffered from severe pneumonia caused by MRSA in early postoperative stage. It was effective to give antibiotics and gamma-globulins. PMID- 8409622 TI - [A case of subpulmonary membranous stenosis associated with atrial septal defect]. AB - The patient was a 5 years old male who had had the cyanosis and congestive heart failure from his neonatal period. Dopamine, digitalis and diuretics disappeared his symptoms and he had been followed up as the out-patient. Preoperative cardiac catheterization revealed atrial septal defect and moderately pulmonary stenosis with two-staged systolic pressure gradient in a right ventricular cavity. Right ventriculogram showed subpulmonary crescent-shaped, linear filling defect. Ventricular septal defect was not detected. He was underwent open heart surgery and subpulmonary membranous stenosis was found out. Pressure gradient across the right ventricular outflow tract was diminished by the resection of the membranous structure. Atrial septal defect without lower margin was closed directly. Postoperative course was uneventful. Right ventricular apical systolic pressure was decreased to the degree of 27 mmHg postoperatively. PMID- 8409623 TI - Achieving irreducibility of the Markov chain Monte Carlo method applied to pedigree data. AB - Markov chain Monte Carlo (MCMC) methods have been explored by various researchers as an alternative to exact probability computation in statistical genetics. The objective is to simulate a Markov chain with the desired equilibrium distribution. If the transition kernel is aperiodic and irreducible, then convergence to the equilibrium distribution is guaranteed; realizations of the Markov chain can thus be used to estimate desired probabilities. Aperiodicity is easily satisfied, but, although it has been shown that irreducibility is satisfied for a diallelic locus, reducibility is a potential problem for a multiallelic locus. This is a particularly serious problem in linkage analysis, because multiallelic markers are much more informative than diallelic markers and thus highly preferred. In this paper, the authors propose a new algorithm to achieve irreducibility of the Markov chain of interest by introducing an irreducible auxiliary chain. The irreducibility of the auxiliary chain is obtained by assigning positive probabilities to a small subset of the genotypic configurations inconsistent with the data, to bridge the gap between the irreducible sets. PMID- 8409624 TI - A field model of left-right asymmetries in the pattern regulation of a cell. AB - Dynamical models can offer insights into important aspects of patterns that recur throughout the biological world, such as global symmetries, mirror images, and pattern reversals, etc. Building on phenomenological rules of pattern formation (e.g. E. M. Nelsen and J. Frankel, 1986, Dev. Biol. 114, 53), the author developed a mathematical model of a morphogenetic field, expressed as a vector field, which contains the information to specify a pattern. The dynamics of the field arises from the minimization of a nonlinear energy density functional, a quartic polynomial of the field together with field gradient effects. This model was successfully applied in detail to surface patterns observed in right-handed Tetrahymena, where reproducible regions of left- and right-handed domains appear during regulation. Counterpart experiments on left-handed patterned cells suggest an intrinsic asymmetry between right- and left-handed patterning in this biological system, causing left-handed cells to follow different regulation pathways from the right-handed cells. We show that the energy minimization approach can account for this globally different behaviour in a version of the model with small local asymmetry. PMID- 8409625 TI - Modelling the spread of HIV among intravenous drug users. AB - This paper considers a random allocation model for the transmission of HIV by needle sharing among a group of intravenous drug users who are friends or relatives (buddy-users). A Markov chain approach is used to track the increase in infectives in a stable group of such IVDUs, some of whom are HIV positive. The model is modified to allow for the replacement of infectives in the group, with the group size remaining constant. Further problems are suggested. PMID- 8409626 TI - The stochastic general epidemic model revisited and a generalization. AB - While the mathematical theory of epidemics has its origins with Ross (1911), it was not until Kryscio (1975) that explicit expressions for the state probabilities of the classical general epidemic model established by Bartlett (1949) were found. However, these formulae were of limited practical use when the population size was of even moderate size. By shifting the focus from the bivariate pair representing the number of susceptibles and infectives to that for the number of infectives and removals, one is able to obtain solutions that are considerably simpler and easier to manage than those previously derived and which are not restricted by the size of the population. The results are obtained for a generalized general epidemic process in which transition probabilities are arbitrary functions of the state space, and then applied to the classical model. An extension to time-dependent transition rates is also considered. PMID- 8409627 TI - Thigh pain in cementless total hip replacement. AB - The incidence of thigh pain was investigated in 51 joints implanted with model Y or YII cementless total hip system. The 51 patients were followed for one or more years after surgery. Twenty-six out of 51 implanted with the model Y, and the other 25 with the model YII. Thigh pain appeared in 11 out of 26 patients (43%) replaced by model Y and in only 2 out of 25 (8%) replaced by model Y II. The difference in the incidence of thigh pain was attributed to the difference in the extent of the press fit of the stem, stress distribution and postoperative activity. To prevent thigh pain, a favorable press fit must be achieved during surgery, and the excessive weight-bearing must be restricted in postoperative course. In our improved stem design, stress is distributed uniformly on the interface between the bone and the stem resulting in reduction of thigh pain. PMID- 8409628 TI - [One stage operation for dead space of pyogenic and tuberculous osteomyelitis]. AB - Pyogenic and tuberculous osteomyelitis is known to have difficult surgical problems. Complete surgical excision of dead and grossly infected soft tissues and osseous structures frequently results in large defects. Hydroxyapatite filler has been used to pack osteomyelitic defects. Between 1984 and 1991 we have treated 6 foci in 6 patients: 4 males aged 61, 54, 47 and 26 and; 2 females aged 59 and 36 years. There were 3 cases of chronic osteomyelitis, and one each of acute osteomyelitis, tuberculous osteomyelitis, and Brodie's abscess. All were reviewed within a period of between 21 to 94 months postoperatively. The pathogens cultivated were Mycobacterium tuberculosis in case 2 and Staphylococcus aureus in case 3 and case 6. In other cases, cultures revealed no pathogens. The hydroxyapatite fillers were mixed with an antibiotic. In addition, in the last four cases the fibrin sealant and CaCl2-thrombin solution were mixed. The excavated bone defect was packed with this composite biomaterial. Neither closed irrigation/suction using antibiotic solution nor a cast was used. Antimicrobial therapy directed specifically to the deep tissue specimens, was administered to all patients. On examination, all of the foci had completely healed by the end of the follow-up period. The cure of pyogenic and tuberculous osteomyelitis is obtained from obliterating dead spaces, which prevent the survival of organisms. Hydroxyapatite is the biomaterial most compatible with human bone and is also suitable for obliterating dead space. The advantages of the fibrin sealant system are that defects can be packed without gaps. Thus, postoperative hematomas can be avoided. In addition, the antibiotic incorporated into the fibrin sealant is released more slowly from the concentrated fibrin than from hematomas. As a result, pathogens multiply more slowly in fibrin. From these findings, we believe that this new method is simple, can be performed safely in one stage, and offers satisfactory results. PMID- 8409629 TI - [Bone histomorphometric study of the ilium in psychiatric patients with longterm administration of anti-psychiatric drugs]. AB - Histomorphometric studies were performed on biopsies of the spongiosa from the iliac crest of patients with fracture who had been taking medication for a psychiatric condition. The biopsy had been performed in 30 individuals, 10 of which were males and 20 females, ages ranging from 23 to 63 years, and from 21 to 75 respectively. Twenty healthy individuals were use as control. All anti psychiatric drugs in this study have been calculated in terms of chlorpromazine (CPZ). The purpose of the study and intraoperative biopsy had been explained to the patients and their agreements were obtained before the surgery. The undecalcified specimen made from the iliac crest were studied using a semiautomatic image analyzer. In the result of this study, there was decrease in the bone volume (BV/TV), the osteoid surface (OS/BS), and the eroded surface (ES/BS) in the affected subjects compared to the controlled group. However, there was increase in the relative osteoid volume (OV/BV) among the subjects who had had a psychiatric condition for 21 years or more and taking more than 500 x 10(4) mg of CPZ. There was no significant difference in the mineral apposition rate (MAR) between the dosage of CPZ and periods of intake; however the MAR was elevated in the affected group as compared to the controlled group. In conclusion, prolonged administration of CPZ may increase the volume of osteoid and disturb the calcification. PMID- 8409630 TI - [The respiratory movement of rib cage in relation to electromyographic activity of the biceps brachii muscle neurotized by the intercostal nerves]. AB - The motor unit potentials of biceps brachii muscle innervated by intercostal nerves and those of second intercostal muscle in relation to respiratory cycle were studied by electromyography (EMG) in 23 patients with traumatic brachial plexus palsies. These patients were followed from about 2 years to 17 years averaging 5 years after intercostal nerves crossing to musculocutaneous nerve. The integrated EMG of biceps brachii muscle ranged from 4 muVS to 244 muVS per second during respiratory phase, with voluntary elbow flexion weighted 500 grams in the hand. Twelve cases showed no significant differences of integrated EMG of biceps brachii muscle between the inspiratory and the expiratory phase. Eleven cases exhibited a significant difference (p < 0.05) between the inspiratory and the expiratory phase. Four out of 11 cases displayed much more integrated EMG activities of biceps brachii muscle in the expiratory phase than those in the inspiratory phase; all these four showed excellent biceps brachii muscle strength. Departure of voluntary elbow movement from the respiratory cycles did not always give the biceps brachii muscle satisfactory strength and did not relate to the postoperative time after nerve surgery. PMID- 8409631 TI - [Role and movement of the toes during walking]. AB - In order to understand the function of toes during free walking, movements of the toes of 30 normal adults were measured with the electrogoniometers. The angular changes of the interphalangeal (IP) joint of the 1st toe and the distal interphalangeal (DIP) joints of the 2nd to 4th toes, the ground reaction force, foot switch signal and the pressure changes in the forefoot were recorded simultaneously. The relationships between these measurements were then investigated. During the foot-flat phase, the IP joint of the 1st toe stayed at a flexed position, while the DIP joints of the 2nd to 4th toes remained at a slightly extended or neutral position but there was scarcely any pressure on the toe tips (0-0.3 kg/cm3). After the heel-off, the pressure on the 1st toe tip gained with increasing flexion of the IP joint of the 1st toe. The mean of the maximum angles was 13 +/- 5 (mean +/- SD) degrees. After the heel-off, the DIP joints of the 2nd and 3rd toes remained in the extended position in 16 cases (6.6 +/- 3.1 and 6.3 +/- 3.4 degrees) and appeared to stay at a flexed position in 14 cases (10.0 +/- 7.8 and 8.0 +/- 5.3 degrees). The DIP joint of the 4th toe remained in the extended position in all cases (9.2 +/- 8.5 degrees). The pressure of the toe tips increased. The magnitude of the pressure of the extended toe tips was higher than that in the flexed toe tips. The pressure of the toe tips of the 1st to 4th toes reached the maximum much earlier than that of the toe off. From these observations, we conclude that the function of the toes during free walking seems to stabilize the foot by extending a contact area with floor mainly after the heel-off. PMID- 8409632 TI - [Dynamic analysis in the knees with chronic anterior cruciate ligament insufficiency--an evaluation of antero-posterior instability, leg rotation and ground reaction force]. AB - A dynamic analysis was made on the knees with chronic anterior cruciate ligament (ACL) insufficiency for antero-posterior instability and abnormal rotation, also evaluating them for ground reaction force and muscle strength of knee extension. Studies were carried out on 51 patients with chronic unilateral ACL insufficiency and 80 knees of 40 healthy male and female young adults as controls. Using a knee motion analyser, an apparatus designed to analyse three dimensional knee motion, the gait was studied on a force plate. At the same time, the muscle strength of knee extension was measured with a Kinetic-Communicator (KIN-COM). In the dynamic analysis of the knee motion anterior instability was notable at a small angle of flexion. Qualitative evaluation of the knee motion revealed three patterns of rotation. The evaluation of ground reaction force showed that the rise from the heel strike was slow, its slope was gentle and the effect of weight removal was unclear. The evaluation of the muscle strength of knee extension revealed a decrease in torque of muscular contraction at 20 degrees of knee flexion. PMID- 8409633 TI - [Electrophysiologic analysis of the lumbosacral radiculopathy using nerve root conduction velocity (NRCV) and cauda equina action potentials (CEAP)]. AB - Nerve root conduction velocity (NRCV) and cauda equina action potential (CEAP) have been measured to assess the severity of lumbosacral radiculopathy, the level specific diagnosis of the symptomatic roots, and to predict the outcome. This study included 71 patients (40 males, 31 females, average age of 54 years at the time of surgery) who underwent decompressive surgery for lumbar radiculopathy. The NRCV and CEAP were directly measured during the operation. The NRCV decreased significantly with progression of radicular symptoms. The NRCV showed a marked reduction in the nerve roots of the patients with a two years or longer history of radicular symptoms; or those with compression of the nerve roots on the imaging examinations; or nerve roots that were considered to have been subjected to persistent compression over a prolonged period with severe inflammation and adhesions. Multivariative analyses suggested that the NRCV correlated closely to the postoperative neurologic recovery, and the outcome of the lumbosacral radiculopathy could be predicted to some extent by measurements of NRCV. The level-specific diagnosis of the radiculopathy could be determined when the CEAP showed a more than 30% left-right potentials difference. PMID- 8409634 TI - [Thoracolumbar burst fractures; an experimental study on cadaveric spines and finite element method]. AB - A biomechanical study on cadaveric human and calf spines and finite element method (FEM) was carried out in order to detect the mechanism of thoracolumbar burst fractures and to investigate the influence of disc degeneration and bone mineral density (BMD). Eleven motion-segments of thoracolumbar spines from human cadavers and 5 ones from calf thoracolumbar spines were compressed vertically until a fracture occurred. Typical burst fracture occurred in seven of eleven human specimens and in all calves spines, which showed destruction of the middle end-plate and disc materials in the vertebral body. The fracture line was located between the middle of the end-plate and the middle of the posterior wall cortex. No burst fractures could be seen in the specimens with severely degenerated disc and osteoporosis. Finite element analysis corresponded to the results of experimental study. Therefore, the axial compression proved to induce typical burst fractures in the thoracolumbar spines. Mechanism of this fracture was influenced by bone mineral density and disc degeneration. PMID- 8409635 TI - Ultrastructural changes of collagen fibers in the anterior cruciate ligament of bipedal rats after enforced running. AB - The effect of enforced running on the morphological characteristics of the fibroblasts and on the distribution of the collagen fibril diameter in the anterior cruciate ligament of bipedal rats was examined by electron microscopy. Twelve bipedal rats were divided into two groups of equal numbers: (1) those without exercise, and (2) those with enforced exercise on a treadmill for four weeks. Ninety percent of the fibroblasts in the rats without exercise were ovoid shaped with a round nucleus, whereas eighty-one percent of those in the rats with exercise were spindle-shaped with indented nucleus. There was a significant difference in the ratios of ovoid-shaped to spindle-shaped cells between the two groups (p < 0.05). There was an increased number of rough surfaced endoplasmic reticula and lysosomal vesicles in the fibroblasts of the rats with exercise. The collagen fibril diameters of the rats with exercise varied in size from 20 to 120 nm, while those of the rats without exercise were uniform in size, mostly between 50-80 nm. Mean collagen fibril diameter was 70.8 +/- 16.6 nm in the rats without exercise, and 66.6 +/- 21.9 nm in the exercised rats. This difference was significant (p < 0.05). These electron-microscopical findings indicate an increased collagen metabolism in the anterior cruciate ligament of rats with enforced running. PMID- 8409636 TI - [An experimental study on injuries to the growth plate--effects on the growth of long bones following placement of smooth pins across the growth plate of immature rabbits]. AB - Displaced epiphyseal separation is difficult to reduce and fix. We usually pass a few Kirschner wires from the epiphysis to the metaphysis to maintain the position of reduction. However there are various opinions about the effects of penetrating smooth pins, concerning whether it causes retarded bone growth. In this experimental study on immature rabbits, stainless steel pins 0.7, 1.2, 2.0, or 3.5 mm in diameter or a single crystal alumina ceramic pin 1.2 mm in diameter were driven into a drill hole of equivalent bore in the intercondylar portion of the right femur across the central portion of the growth plate. Using this model, we analyzed effects on the growth plate and bone growth macroscopically, radiographically and histologically. The follow-up intervals were 3, 6, 12 and 24 weeks. At 12 and 24 weeks after insertion of the 3.5 mm stainless steel pin, shortening of the operated femur was observed. In other groups there was no difference in the length of the femur. The pin position after insertion was divided into two types; in one the pin position remained located in the growth plate, and in the other the pin migrated proximally beyond the growth plate. Histologic studies showed regenerated growth cartilage in the implant canal at the plane of the growth plate. At 12 weeks after insertion of a 3.5 mm stainless steel pin there was bone bridge between epiphysis and metaphysis without clear regenerated growth cartilage. The role of regenerated growth cartilage, therefore, appears to prevent bone bridge formation. There was no significant difference between the stainless steel pin and the single crystal alumina ceramic pin. When the diameter of the pin is sufficiently large and also low growth plate activity due to aging is present, retarded bone growth would be anticipated. PMID- 8409637 TI - Meniscal transplantation using fresh and cryopreserved allografts--an experimental study in the genetically defined rat. AB - The purpose of this study is to examine the possibilities of meniscal allograft and the effects of cryopreservation on grafts in rats (Lewis and Fisher). Transplants were carried out on three minor mismatched groups of rats with: 1) 16 fresh meniscal allografts, 2) 16 fresh meniscal allografts with immunosuppression, and 3) 16 cryopreserved meniscal allografts. As a control, 16 Fisher rats received isograft menisci transplants. Four transplanted grafts each were evaluated histologically 4, 8, 12 and 21 weeks after grafting. Isograft meniscal transplantations were judged to have been successful. At 21 weeks, all of the fresh allografts had survived in the immunosuppressed rats, however, without immunosuppression, virtually all fresh allografts became scar tissue. On the other hand, all cryopreserved meniscal allografts survived; three out of the four grafts showed no degenerative changes 21 weeks after transplantation and produced the best results in those rats without immunosuppression. PMID- 8409638 TI - [Basic and clinical knowledge of methicillin resistant Staphylococcus aureus]. PMID- 8409639 TI - [Proceeding of the 8th Annual Meeting for the Orthopaedic Research of the Japanese Orthopaedic Association. Matsumoto, October 7-8, 1993]. PMID- 8409640 TI - [Radiological study of the morphological abnormalities on lumbosacral spine--with special reference to clinical features]. AB - The relationship between clinical features and radiological findings in patients with morphological abnormalities including lumbosacral transitional vertebrae (TV), spina bifida occulta (SBO), and lumbar spondylolysis (LY) was investigated. Materials were 1194 cases including 408 TV, 402 SBO, 162 LY, 85 LY+SBO, 41 LY+TV and 96 SBO+TV. Five hundred cases showing normal structure of lumbosacral spine in X-rays were also studied as a control group. Many of the cases with clinical symptoms showed the low position of the intercrestal line, 34.4% of the patients with incomplete diarthroial joint of TV complained of sciatica or numbness of the leg. The range of motion of the vertebral body directly above TV increased in patients with TV located below the intercrestal line. In many cases of the 5th lumbar spondylolysis, a decrease of the lumbosacral angle and an increase of the pedicle facet angle in the 4th and 5th lumbar vertebrae were observed. In many cases of co-existence of the 5th lumbar spondylolysis and SBO in the 5th lumbar vertebrae, it was observed that a decrease of the lumbosacral angle and the lumbar index and an increase of the pedicle facet angle compared with cases of the 5th spondylolysis. PMID- 8409641 TI - [Effect of 1 alpha-OH-D3 on the bone metabolism after high tibial osteotomy]. AB - The effect of 1-hydroxycholecalciferol (1 alpha-OH-D3) on bone metabolism during osteotomized bone healing was investigated. The study based on 19 female patients with gonarthrosis who underwent the high tibial osteotomy. In 10 selected randomly from these patients 1 alpha-OH-D3 (1.0 micrograms/day) was administered before and after the surgery. After operation the changes in bone density (by MD method) and biochemical parameters were followed. In the non-treated group the serum level of [Ca] x [P] product temporarily decreased at 3 days after surgery. Average bone density at two months after the surgery was 8.8% lower than the preoperative value. In contrast, 1 alpha-OH-D3 treated group showed significantly less decrease of serum [Ca] x [P] product at the third day than the control group and no significant decrease of bone density at two months after the operation. From these results, it was concluded that the 1 alpha-OH-D3 suppresses the decrease of serum Ca and P level and systemic osteopenia after osteotomy and may have an beneficial effect on fracture healing. PMID- 8409642 TI - Natural history of advanced osteoarthritis of the hip. AB - We followed long-term osteoarthritis of the hip clinically and radiographically in 38 patients (47 hips) finally reaching an advanced or terminal stage for more than 10 years. The relation between various measured parameters and disease progression and osteophyte formation was analyzed. By multivariate analysis, the clinical outcome of osteoarthritis of the hip could be correctly predicted in 91.5% of cases. In the cases in which initially the slope of the acetabular roof was not steep, the greater trochanter was low, and the cranial joint space was narrow, good roof osteophyte formation was well recognized. The cases with good roof osteophyte formation were characterized by preserved roundness of the femoral head, no shortening of the femoral neck, no lateralization of the femoral head, unchanged cranial joint space, and no worsening of pain at the final follow up. PMID- 8409643 TI - [Isokinetic torque of quadriceps in patients with untreated anterior cruciate ligament injury of the knee joint]. AB - In order to study the role of muscle strength in patients with untreated anterior cruciate ligament (ACL) injury of the knee joint, the torque of the quadriceps was isokinetically measured during concentric contraction (CC) and passive eccentric contraction (PEC). The results were compared with those in normal individuals and sports players. Ninety patients with untreated ACL injury were subjected for the study. There were 50 men and 40 women. The normal group consisted of 20 students and the sports player group consisted of 20 soccer players. The peak torque and the torque at 30 degrees flexion of the quadriceps were isokinetically measured during CC and PEC by using a BIODEX machine. RESULTS: In patients with ACL injury, the peak torque was smaller in the injured side as compared with the uninjured side. At the same angle velocity, decrease of PEC in the injured side was larger than that of CC. These results were compared with the normal group and the sports player group. There was significant difference in the peak torque per body weight among each groups. But, the torque per body weight at 30 degrees flexion was almost consistent in these three groups. DISCUSSION: A considerable quadriceps atrophy occurs following ACL injury. In our experience, patients with functional absence of ACL are much more difficult to return to vigorous sports which require explosive eccentric contraction than to return to endurance sports which require repetitive concentric contractile efforts of the quadriceps. In the present study, decrease of the passive eccentric contraction in the injured side was more pronounced than that of the concentric contraction. From these results, the significantly decreased passive eccentric torque in the patients with untreated ACL injury can be a factor to cause instability of the knee joint in active daily life as well as in sports activities. PMID- 8409644 TI - [A biological and morphological study of ovoid and spindle cells in malignant fibrous histiocytomas]. AB - To clarify the histogenesis of malignant fibrous histiocytoma (MFH), ovoid and spindle cells, the major components of this tumor, were biologically and morphologically studied using the following methods: (1) flow cytometric nuclear DNA analysis, (2) argyrophil staining for nucleolar organizer regions (Ag-NORs), (3) separation of MFH cells into the two types described above using a flow cytometric cell sorting system, (4) electron microscopic morphological observation, and (5) immunohistochemical examination. Specimens were obtained from 24 cases of soft-tissue MFH and 7 cases of MFH in the bone, which were treated in our institute. In ovoid cells, aneuploid pattern was shown by nuclear DNA analysis and high proliferation activity was found on Ag-NORs staining. Ultrastructural analysis showed that the ovoid cells had malignant characteristics and were presumedly histiocytic in origin. Immunohistochemical examination also, supported histiocytic origin. However, spindle cells were not identified as malignant tumor cells by the various methods described above. Although the histogenesis of MFH remains to be elucidated, these findings provide interesting information. PMID- 8409645 TI - [Measurement of the center of gravity using the link model for controlling functional electrical stimulation]. AB - The purpose of this study is to better understand the reliability of using the link model when determining the center of gravity. We measured the centers of gravity using both the link model and the floor reaction force in 11 healthy subjects. Dempster's and Clauser's anthropometric data were used in measuring the center of gravity by the link model, and these results were compared with those measured by the floor reaction force. The motion area of the center of gravity during quiet standing was determined by experimental data, using the link model calculation. Joint torque was measured using Cybex II in one paraplegic patient who was treated by functional electrical stimulation (FES). We discuss whether the paraplegic patient can stand safely in a determined center of gravity motion area. Displacements of the center of gravity measured by the link model were very close to those obtained by the floor reaction force. The parameters of body segments proposed by Dempster were more reasonable than those of Clauser. The maximum differences of the center of gravity between those measured by the link model using Dempster's data and those measured by the floor reaction force, were 1.47 cm for the fore-aft direction, 1.85 cm for the lateral direction, and 2.34 cm for the vertical direction. The torque caused by electrical stimulation was stronger than that calculated by the link model during quiet standing. Our results suggest that the link model system that measures displacement of the center of gravity using Dempster's data, is clinically applicable for the closed loop control system of FES. PMID- 8409646 TI - Immuno-reactive human epidermal growth factor (h-EGF) in rheumatoid synovial fluids. AB - The immuno-reactive human epidermal growth factor (h-EGF) level, measured by a sensitive enzyme immunoassay, was significantly higher in synovial fluids or synovial tissue extracts from 89 patients with rheumatoid arthritis (RA) than in those from 53 patients with osteoarthritis. RA synovial immuno-reactive h-EGF was predominantly of a low molecular weight form (LMW h-EGF) on gel filtration chromatography. Furthermore, in the RA group, the synovial immuno-reactive h-EGF correlated positively with C-reactive protein, an acute-phase reactant, in the blood examination and with synovial immunoglobulin M. These results suggest that synovial h-EGF is specifically produced by RA synovium from the initial stage of arthritis, and that the measurement of synovial h-EGF serves as an early indicator to appraise the RA activity. A pathogenesis of RA including growth factors and growth inhibitory factors are discussed. PMID- 8409647 TI - [Theory and practice of therapeutic exercise in orthopedics]. PMID- 8409648 TI - Management of fingertip injuries. PMID- 8409649 TI - Fingertip injuries management with semi-occlusive dressing. AB - We have developed a simple, cheap and efficient method of management of fingertip injury using a semi-occlusive dressing ("Opsite"--Smith and Nephew). The fingertip is covered with the "Opsite" once a week only. The dressing provides a temporary "skin", making the finger painless. This semi-occlusive "skin" allows the healing environment to reach an optimal milieu (e.g. pH, oxygen, tension, immunoagents) actively promoting granulation tissue formation and epithelialization. The result of 200 fingertip injuries treated with this method proves the development of a near normal pulp shape and useful epithelium within an average of 20 days. PMID- 8409650 TI - The lateral pulp flap. PMID- 8409651 TI - Arterial anatomy of the thumb. AB - The anatomical literature has indicated that the arterial supply to the thumb comes from the princeps pollicis artery. However, this simplified description does not often correlate with intraoperative findings. The purpose of this study was to investigate and clarify this important area of anatomy by dissection of fresh cadaver hands. 40 dissections were completed on 35 intravascularly injected and five non-injected hands. Five patterns were identified. The most common pattern showed both a superficial and deep vessel to the first web space in 54% of specimens. Dominant vessels included the superficial palmar branch of the radial artery in 8%, first palmar metacarpal artery in 18% and dorsal metacarpal artery in 8%. Only three specimens correlated with the textbook description. We conclude that the term "princeps pollicis" is actually a misnomer. PMID- 8409652 TI - The posterior interosseous flap in primary repair of hand injuries. A review of 23 cases. AB - The posterior interosseous flap has been used for resurfacing in 23 cases of hand injury in the past 5 years. There was complete necrosis in two cases, partial necrosis in three and temporary post-operative nerve palsy in one. PMID- 8409653 TI - Intra-operative tunnel expansion to prevent tunnel compression following latissimus dorsi muscle transfers. AB - The intra-operative tunnel expansion technique is described to prevent tunnel compression following muscle transfers. The results of ten cases of latissimus dorsi transfer using this new technique are reported. PMID- 8409654 TI - Repair of digital nerve defect with autogenous vein graft during flexor tendon surgery in zone 2. AB - Autogenous vein graft was used to fill 18 digital nerve defects between 0.5 to 5.8 cm in length during flexor tendon surgery in zone 2. The vein was taken from the forearm and reversed to bridge the digital nerve. For nerves with defects over 2.0 cm, normal nerve slices were inserted inside vein conduits. Recovery of sensibility was evaluated by von Frey test, pin-prick detection, localization of stimulus, moving two-point discrimination and sweating on the finger pulp. Follow up revealed excellent recovery in two digital nerves, good in nine, fair in five and poor in two. The results suggest that vein graft provides a simple and practical method to reconstruct a digital nerve defect during tendon repair in zone 2. PMID- 8409655 TI - Intrinsic reinnervation--myth or reality? AB - A clinical and electromyographic study of major nerve transections at the wrist in 22 patients has shown that electrical recovery nearly always occurs in the intrinsic muscles, despite the absence of clinically detectable function. There appears to be a level of electrical reinnervation above which clinically detectable intrinsic muscle power is usually present. This level would appear to be 50% of the contralateral maximum evoked muscle action potential; above this the "myth" of clinical recovery becomes a reality. PMID- 8409656 TI - Basil Kilvington. Unknown pioneer of peripheral nerve repair. AB - In the early years of the present century, a group of experiments assessing the results of the surgical repair of peripheral nerves and spinal roots was performed by Basil Kilvington. The outcome of the experiments was assessed using both electrophysiological and morphological techniques. Much of Kilvington's work remained unrecognized and was thus repeated at a later date. Kilvington's role in the early history of the surgical repair of nerves appears to have been forgotten and his substantial and prophetic discoveries deserve better recognition. PMID- 8409657 TI - Entrapment neuropathy of the palmar cutaneous nerve within its tunnel. AB - A case of isolated entrapment of the palmar cutaneous branch of the median nerve is presented. At operation, a ganglion compressing the nerve within its tunnel was found. Symptoms were relieved with no recurrence at 2 years after decompression and excision of the ganglion. PMID- 8409658 TI - Surgical management of recurrent carpal tunnel syndrome. AB - An approach to the surgical management of recurrent carpal tunnel syndrome was evaluated in 30 patients with 35 involved wrists. This includes internal neurolysis of the median nerve and early post-operative mobilization of the wrist and fingers. The preferred surgical approach is through a second, more ulnar incision. Clinical assessment of sensorimotor function was converted into a numerical score ranging from zero (normal) to 9 (anaesthesia) and 10 (atrophy, severe). The average pre-operative score was mean 6.5 and median 7. At a mean follow-up of 23.5 months, the average post-operative score was mean 1.8 and median 0, a statistically significant improvement (P < 0.001). PMID- 8409659 TI - Long-term results of carpal tunnel decompression. Assessment of 60 cases. AB - The long-term prognosis of 60 patients operated on for carpal tunnel syndrome has been assessed in a prospective study with a median follow-up period of 5.5 years (range 2-11 years). Analysis of motor, sensory, trophic, and electrodiagnostic findings and assessment of pain were performed pre- and post-operatively using a standardized grading system. The results were generally favourable with a variable degree of improvement in 86% of cases. Statistical evaluation using multiple Dunn-Rankin tests revealed pain to be the most prominent pre-operative finding. On post-operative re-examination, pain was found to be improved to a significantly greater extent than any other variable. Analysis of several potential prognostic factors showed that pain lasting for more than 5 years prior to surgery indicates a poor prognosis. Only patients with diabetes mellitus exhibited a trend toward less pain relief. PMID- 8409660 TI - Median nerve compression following trauma in children. A report of two cases. AB - Two cases of subacute median nerve compression in children following trauma are presented. The first highlights problems of incised wounds around the wrist. The second illustrates an unusual presentation of nerve compression following a blunt injury. PMID- 8409661 TI - Cutaneous malignant peripheral nerve sheath tumour (MPNST) of the hand: a review of current literature. AB - Malignant peripheral nerve sheath tumours (MPNST) are spindle cell sarcomas normally situated in the deep soft tissues. Cutaneous MPNST is an uncommon variant, usually occurring in the head and neck. When it arises in the upper limb this tumour may pose a considerable diagnostic and therapeutic problem. We present a case of cutaneous MPNST arising in the hand. Tumour extension was exclusively perineural along three major nerve trunks from an interdigital origin. Current knowledge of the clinical behaviour of cutaneous MPNST is reviewed. PMID- 8409662 TI - Magnetic resonance imaging of an obscure glomus tumour in the fingertip. AB - Magnetic resonance imaging was performed on a patient with chronic obscure pain in her little finger. MRI findings helped to establish the diagnosis and guide the surgeon to the glomus tumour. PMID- 8409663 TI - Skin tumours of the hand. A 10-year review. AB - A 10-year retrospective study was performed in order to determine the incidence, distribution, histological type and behaviour of skin tumours of the hand that were referred to a regional Plastic Surgery unit. 85 patients were studied and 98 malignant or pre-malignant lesions identified. The majority were squamous cell carcinomas in male manual workers. Recurrence after excision was seen only in lesions greater than 1.5 cm in diameter. The overall incidence of squamous cell carcinoma of the hand (requiring surgical excision) was seen to be five cases per million per year. Other skin tumours were rare. PMID- 8409664 TI - Demineralized bone powder. Clinical applications for bone defects of the hand. AB - The results of 20 consecutive cases of bone defects of the hand treated by curettage and implantation of demineralized bone powder compare well with a similarly matched retrospective group managed with autogenous grafts and curettage alone. Results revealed "excellent" (16 patients) or "good" bone healing (4 patients) in the bone powder group on independent radiographic evaluation. Bone bridging was noted at an average of 9.9 weeks. All defects healed without recurrence, resorption or refracture, with follow-up of 4 to 36 months. In the conventionally managed retrospective group there was a 25% failure rate necessitating re-operation. Demineralized bone powder provides a rapid, safe and effective method of management of bone defects of the hand. PMID- 8409665 TI - Bone infection resembling phalangeal microgeodic syndrome in children. A case report. AB - Phalangeal microgeodic syndrome of childhood is a rare condition and is the result of necrosis and repair within the phalanges. The cause is unknown. We present a case in which Brucella melitensis was grown from one of the lesions. PMID- 8409666 TI - Day care surgery for Dupuytren's contracture. AB - Contrary to standard practice in the United Kingdom, primary surgery for Dupuytren's contracture can be performed safely on a day care basis provided that strict criteria are followed. Although demanding on Consultant surgical time, this policy offers a considerable saving in hospital resources. PMID- 8409667 TI - Fowler's tenotomy for mallet deformity. AB - 35 patients with established mallet finger deformities were treated with Fowler's tenotomy of the central extensor tendon. The mean lack of extension before operation was 45 degrees. 26 patients regained full extension, eight patients had a residual deformity of 10-20 degrees and one patient of 30 degrees. PMID- 8409668 TI - Rupture of the ulnar collateral ligament of the thumb in a 5-year-old girl. AB - Injuries to the metacarpophalangeal joint of the thumb are rare in children and usually involve avulsion of the bone or cartilage. We describe a tear of the ulnar collateral ligament of the thumb with minimal symptoms in a 5-year-old girl. PMID- 8409669 TI - Distal radio-ulnar ligament motion during supination and pronation. AB - The dorsal and palmar distal radio-ulnar ligaments (DRUL) play an important role in the stability of the distal radio-ulnar joint (DRUJ). Various authorities, however, hold opposite opinions regarding DRUL motion during DRUJ pronation and supination, thus implying opposite techniques for reconstruction of the unstable DRUJ. With the hypothesis that relative displacement would increase in the dorsal DRUL during pronation and would increase in the palmar DRUL during supination, measurements were made of the relative DRUL displacement with a Hall-effect displacement transducer during DRUJ pronation and supination in six fresh cadaver wrists. The hypothesis was confirmed that the dorsal radio-ulnar ligament undergoes relative displacement during pronation, while the palmar radio-ulnar ligament undergoes relative displacement during supination. PMID- 8409670 TI - Patterns of hand fractures and dislocations in a district general hospital. AB - In a retrospective survey of patients with fractures and dislocations attending the Accident department of a District General hospital, comparison was made between patients with fractures and dislocations in the hand and fractures and dislocations of other sites. Patients with hand injuries accounted for 28% of the total patients seen. They were more likely than other fracture patients to be male and between the ages of 10 and 40 years. Almost a quarter of hand injuries attended on Monday. Leisure activities outside the home formed the commonest aetiological factor. Only 66% of patients with hand injuries attended the Accident department within 24 hours of injury. The little finger was the commonest site of injury. This survey emphasizes the differences between hand injuries and injuries of other sites, identifies the risk factors for hand injuries and has implications for the management of hand injuries in a District General Hospital. PMID- 8409671 TI - Spiral fracture fixation techniques. A biomechanical study. AB - The mechanical strengths of five common fixation techniques for spiral fractures have been tested. A total of 240 cadaver metacarpals and proximal phalanges were fractured and fixed by either crossed K-wires, interosseous loops, a dorsal mini plate, a single compression screw or K-wire plus cerclage wire. Specimens were subjected to torsional and cantilever bending tests. A single compression screw provided the best overall fixation for the proximal phalanx. In addition, a single compression screw provided better fixation than any of the other techniques when proximal phalanges and metacarpals were subjected to torsional tests (P < 0.05). In apex dorsal bending tests of metacarpals, the screw provided fixation superior to interosseous wires, crossed K-wires, or dorsal mini-plates (P < 0.05). These results indicate that the use of a single compression screw provides the most satisfactory biomechanical advantage for spiral fracture fixation. PMID- 8409672 TI - Hand injuries from inflation of an air bag security system. PMID- 8409673 TI - Bennett's fracture combined with fracture of the trapezium. A report of three cases. AB - Three cases are reported in which open reduction and internal fixation were required to stabilize an unstable first carpometacarpal joint with simultaneous fracture of the trapezium and Bennett's fracture. The results were good in terms of range of movement and radiological appearance, all of them returning to normal activities, including heavy manual work. PMID- 8409674 TI - The value of scapho-trapezio-trapezoid arthrodesis combined with "de la Caffiniere" arthroplasty for the treatment of pan-trapezial osteoarthritis. AB - 13 patients, 12 female and one male (mean age 63) with pan-trapezial osteoarthritis were treated by a "de la Caffiniere" arthroplasty of the first carpometacarpal joint, combined with a scapho-trapezio-trapezoid (STT) arthrodesis. 11 could be reviewed at least 3 years after surgery (average 3 years and 8 months). Non-union of the arthrodesis occurred twice, once when no internal stabilization was used, once with the use of the staplizer. No impingement syndrome was recorded, and there was no correlation between the angular position of the scaphoid and mobility of the wrist. Two out of 11 "de la Caffiniere" implants had to be removed. Loosening of the stem is unusual, but the cup remained only perfectly in place in two cases. There was no correlation between radiological and clinical findings and the overall result was good or excellent in 8 out of 13 (62%) of the cases. The grip and pinch strength were comparable to the results of the literature. This method cannot be further recommended because of the high rate of re-operation and the risk to the trapezium which has to be further evaluated. PMID- 8409675 TI - Revision procedures for complications of surgery for osteoarthritis of the carpometacarpal joint of the thumb. AB - Between the years 1980 to 1987, 17 patients had revision procedures for complications of surgery for osteoarthritis of the carpometacarpal joint of the thumb. 12 followed silastic implant arthroplasty, four trapeziectomy and one arthrodesis. The failed silastic arthroplasties were treated by removal of the implant, and soft tissue arthroplasty in eight, revision with another implant in three and intermetacarpal bone grafting in one. Patients with metacarpo-scaphoid arthritis after trapeziectomy were treated by silastic implant arthroplasty in three and fusion in one. Nine of the 17 revision procedures (53%) had good, three (18%) fair and five (29%) poor results respectively. PMID- 8409676 TI - The aetiological significance of work-load in the development of osteoarthritis of the distal interphalangeal joint. AB - The incidence of Heberden's nodes was determined in three groups of Japanese women with different occupations to investigate the role of work-load in their pathogenesis. School cooks (n = 260), each of whom prepared 150 to 450 lunches daily, pre-school cooks (n = 222), each of whom prepared 30 to 80 lunches daily, and municipal employees (n = 298) underwent physical examination and X-ray of the hand if pathology was present. The incidence of Heberden's nodes and X-ray incidence of osteoarthritis was 19.2% and 13.5%, 8.6% and 5.4%, 5.9% and 2.0% among school cooks, pre-school cooks and municipal workers, respectively. The incidence of Heberden's nodes and osteoarthritis was significantly higher among school cooks. These findings support the concept that work-load is an aetiological factor in the pathogenesis of Heberden's nodes and also support Radin's hypothesis that women's daily work pattern explains the increased incidence of DIP joint osteoarthritis in women. PMID- 8409677 TI - An extension to the incision for metacarpophalangeal joint surgery. A V-Y extension of the incision for MP joint surgery in rheumatoid arthritis allows lengthening of the skin on the ulnar side of the hand. PMID- 8409678 TI - A simple technique for removing stubborn rings prior to hand surgery. PMID- 8409679 TI - The standard postero-anterior radiographs show two margins of the distal radius. PMID- 8409680 TI - Natural history of scaphoid non-union. PMID- 8409681 TI - Ideas, thoughts and observations on new aspects of science and medicine. PMID- 8409682 TI - Lack of prognostic value of immunoreactivity for p62 oncoprotein in colorectal carcinoma. AB - Immunohistochemical staining for c-myc p62 oncoprotein (p62) was performed in 59 colorectal carcinomas using mouse monoclonal antibody against human c-myc protein (MYC1-6E10). The immunoreactivity for p62 was intense in 22 (37.3%), and weak in 37 (62.7%) of the tumors. Advanced disease and nodal metastasis were more frequently seen in tumors showing intense staining but the difference did not reach statistical significance. Sixteen patients have died of the disease but overall survival rates did not differ between patients with tumors showing intense staining and those with tumors showing weakly staining. In 50 patients undergoing grossly curative resection, local or distant metastasis developed in 15 patients. Immunoreactivity for p62 had no effect on the rate of recurrence or disease free survival rates in these 50 patients undergoing grossly curative resection. The present study revealed that the immunoreactivity for p62 had no prognostic value in colorectal carcinoma. PMID- 8409683 TI - An evaluation of the effectiveness and safety of razoxane when used as an adjunct to surgery in colo-rectal cancer. Report of a controlled randomised study of 603 patients. AB - A prospective controlled randomised trial to evaluate the effectiveness and safety of razoxane is reported. Some 603 patients with colo-rectal cancer having curative surgery entered the study, and all have been followed up for a minimum of five years. Statistical analysis showed that razoxane treatment had no effect either beneficial or adverse on the rates of recurrence or on five year survival of patients with colo-rectal cancer. It is possible that a more prolonged course of razoxane might have significantly influenced survival. The incidence of severe adverse reaction was low but it is of concern that one patient developed leukaemia. Should razoxane be considered for future use it is recommended that continuous low dose therapy be given for no longer than 12 months. No renal, hepatic, pulmonary or cardiac toxicity was noted. PMID- 8409684 TI - Peri-operative heparin: a possible adjuvant to surgery in colo-rectal cancer? AB - Analysis of data from a randomised trial of adjuvant razoxane involving 603 patients with colo-rectal cancer having curative surgery is reported. The results show that razoxane was ineffective but peri-operative subcutaneous heparin treatment apparently conferred a statistically significant improvement in survival at 5 years, or equivalently a reduction in the risk of death. This beneficial effect is apparent in both razoxane treated and control patients and is not explained by demonstrable differences between heparin and non-heparin treated patients in the distribution of known prognostic factors. Adjustment for these factors slightly increased the apparent magnitude of the beneficial effect. PMID- 8409685 TI - Double versus single stapling technique in rectal anastomosis. PMID- 8409686 TI - Transcaecal ileal diversion in the management of the 'at risk' distal colonic anastomosis. AB - Transcaecal ileal diversion has been used in association with primary resection and anastomosis to defunction an elective distal colonic anastomosis in 10 patients and to allow on-table colonic lavage with subsequent colonic defunction in 11 patients presenting as an emergency with distal colonic obstruction. Post operative wound sepsis occurred in four patients (19%) with a clinical anastomotic leak in one patient. The median hospital stay was 14 (10-19) days. Transcaecal ileal diversion is simple to perform. It may facilitate primary resection and anastomosis in both the elective and emergency situation without increasing morbidity, mortality or the hospital stay. PMID- 8409687 TI - The clinical contribution of integrated laboratory and ambulatory anorectal physiology assessment in faecal incontinence. AB - To determine the clinical value of anorectal physiology testing, we have assessed 73 patients with neurogenic incontinence (median age 55 years, 60 female) and 115 controls (median age 48 years, 81 female). All the faecally incontinent patients displayed abnormal anal mucosal electrosensitivity and prolonged pudendal nerve latencies. Rectal compliance was poor in 14% of patients with neurogenic incontinence. Twenty-seven sphincter injuries were identified by endoanal ultrasonography in patients with neurogenic incontinence. Anal sphincter electromyographic abnormalities were demonstrated in all the incontinent patients. Anal pressures were lower in the incontinent group. Frequent, abnormal internal sphincter relaxations were observed in patients with incontinence during ambulatory assessment. These tests provide objective evidence of injury but do not frequently affect clinical decision making. Endoanal ultrasonography and ambulatory assessment provided clinical information of the mechanism of incontinence in 60% of patients. PMID- 8409688 TI - A regional audit of the investigation and treatment of colorectal and pelvic floor disorders (1984-1991). AB - The activities of a regional physiology unit established for the investigation of colorectal and pelvic floor physiology in health and disease in a clinically relevant setting has been audited and its evolution described over a period of eight years. Trends in surgical treatment of some of these disorders over the same period have also been documented in the Lothian Region. Although there has been little change in the number of patients investigated annually patterns of investigation appear to change. Sphincter manometry, proctometrography and somatosensory reflex measurements have remained the most frequently performed and useful investigations. Spinal stimulation studies increased transiently because of a collaborative investigation of bowel and bladder function in patients with spinal injuries. A considerable increase in surface EMG tests and dynamic proctography has occurred. These trends are thought to be related to interest in defining evacuation dysfunction of the rectum and related problems of the pelvic floor. Isotope proctography now rivals barium videoproctography; at the same time the use of manometric colonic motility studies has diminished. Anal ultrasonography has replaced sphincter mapping in the last year and is being applied to other aspects of anorectal pathology. The last 4 years have seen the introduction and increasing use of non-surgical therapeutic modalities for the treatment of faecal incontinence and constipation: reflex electronic sphincter stimulation, biofeedback and the use of a prokinetic agent to promote colonic motility. Concomitant changes in the surgery of constipation and reconstructive anorectal procedures have been observed. It is recommended that coloproctology units should have easy access to at least one such investigation laboratory within their catchment area together with appropriate auditing facilities. PMID- 8409689 TI - Smoking and ulcerative colitis: a community study. AB - Smoking habits of patients with ulcerative colitis, diagnosed in the City of Cardiff between the 1st of January 1978 and the 31st of December 1987 were examined. There was a significant deficit of current smokers, an excess of life long non-smokers and ex-smokers compared with the general population. Men who had previously smoked presented with colitis later than life long non-smokers (mean age difference 16.1 years). The proportion of ex-smokers in this group of patients with colitis was more than twice that expected in the general population. The interval between cessation of smoking and subsequent onset of colitis in ex-smokers was relatively short and in more than half of them occurred within 8 years. There was no significant difference in the frequency of colectomy or the extent of disease among smokers, ex-smokers or life-long non-smokers with ulcerative colitis. PMID- 8409690 TI - The effect of preliminary bowel preparation on a simple test of colonic transit in constipated subjects. AB - The abdominal distribution of orally ingested radioopaque markers is used to assess total and segmental colonic transit in constipated patients, but interpretation may depend on whether studies are carried out on a full colon or one cleared of faeces. We asked 25 severely constipated patients (age 18-74; 22 F, 3 M) to ingest 50 polyethylene markers (4 mm x 2 mm) at breakfast on 2 occasions 1 month apart. No bowel preparation was used for study 1 but for the second study 2 doses of Sodium Picosulphate (2 x 10 mg) were taken 3 days before ingestion of the markers. All subjects reported a good result which had ceased a day before taking the markers. Marker distribution was assessed by a plain abdominal film taken 72 hours after ingestion. All 25 subjects had more than 50% of markers present at 72 h in study one. Of these 4 showed evidence of outlet obstruction with more than 50% of ingested markers in the rectum. In the remaining 21 subjects markers were distributed throughout the colon in a pattern indicating colonic inertia. Following the administration of purgative there was no significant change in the mean number of markers retained but patterns of marker distribution for individual subjects did alter. Of the 21 patients who previously had colonic inertia, 3 showed outlet obstruction and 1 showed no evidence of delayed transit. Of the 4 patients with a pattern of outlet obstruction in the first study, 2 showed no evidence of delayed transit after Picolax.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409691 TI - The value of exfoliative cytology in the diagnosis of rectal malignancy. AB - This study has assessed the feasibility and reliability of cytology smears taken from patients with known rectal carcinomas without the use of any special instruments. In a pilot study of 23 patients who all had proved rectal cancer, 19 patients had satisfactory smears, one of which produced a false negative result. The remaining 18 displayed varying degrees of dysplasia (10) or frank malignancy (8). One of the patients with a smear containing severely dysplastic cells had a negative biopsy but had clear clinical and radiological evidence of a rectal tumour. Subsequently, a blind controlled study was undertaken on 29 patients with either carcinoma, proctitis or a normal rectum. All 29 sets of smears were of diagnostic quality and were correctly diagnosed as benign or malignant by the cytologist who had no knowledge of the clinical diagnosis. This is a rapid and simple diagnostic test which, if positive for malignancy, allows treatment to be planned at the first consultation. In combination with a forceps biopsy, the rate of positive diagnosis may be increased. A dysplastic smear adds to the level of clinical suspicion and confirms the need for biopsy of a lesion. PMID- 8409692 TI - Anorectal malignant melanoma has a poor prognosis. AB - Clinicopathologic features and surgical treatment of 15 patients with primary anorectal malignant melanoma were studied retrospectively. There was a female preponderance (2:1). The median age was 66 years. Common initial symptoms were rectal bleeding (87%) and/or anal pain (33%); 25% of the melanomas were amelanotic. The maximum tumor size ranged between 0.8 and 8.4 cm (median 3.0 cm). Of the tumors evaluated histologically (n = 12), tumor thickness ranged from 0.9 to 11.3 mm (median 6.1 mm). All melanomas invaded at least into the subepithelial tissue (n = 8) and/or the submucosa of the distal rectum (n = 4), with extension into the internal anal sphincter (n = 5) and lamina propria (n = 3). Endoluminal ultrasound accurately demonstrated depth of invasion in 3 of 3 patients. Three (20%) patients with distant metastases at initial presentation had a mean survival of 8 mo; one of these primary melanomas measured 0.8 cm. Of 12 patients undergoing "curative" treatments--4 by abdominoperineal resection (APR) and 8 by local excision (LE), the incidence of loco-regional recurrence was similar (2/4 and 5/8). All these 7 patients with loco-regional recurrence developed distant metastases within 3 months. The mean survival was similar between APR and LE in the total group (25 mo vs 20 mo), in the decreased (27 mo vs 24 mo) and in those treated with a curative intent (29 mo vs 22 mo). There was no long-term survivor but four patients remained tumor-free up to 19 mo after APR (n = 1) or LE (n = 3).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409693 TI - Turcot's syndrome and familial adenomatous polyposis associated with brain tumor: review of related literature. AB - We investigated clinical manifestations in 124 patients with a possible Turcot's syndrome whose data were taken from documented cases. The cases were subclassified mainly on the bases of the type of familial occurrence and listed in five Tables. We searched for differences in colonic manifestation, histologic type of glioma, mode of inheritance, frequency of parental consanguinity, skin lesions and other accompanying lesions among these five groups. The differences of these clinical findings suggested that glioma-polyposis syndrome should be classified as follows; (1) cases of Turcot's syndrome who had characteristic colonic and brain manifestations, (2) cases of FAP associated with glioma, (3) suspicious cases of glioma-polyposis, and (4) cases other than glioma-polyposis syndrome. PMID- 8409694 TI - How reliable is digital examination for the evaluation of anal sphincter tone? AB - This study investigates whether digital examination of the anal canal can differentiate reliably between competent and incompetent sphincters. Digital examinations were performed by an experienced proctologist who was blinded with regard to the patient's history as well as manometric and endoscopic findings. There was poor correlation between manometric findings and digital assessment of anal sphincter tone. The sensitivity of digital examination for diagnosing incompetent sphincters ranged from 63% to 84%, whereas specificity was calculated as only 57%. It is concluded that even the experienced examiner needs to use more sophisticated methods than digital examination for the evaluation of anal sphincter competence. PMID- 8409695 TI - Anthracycline toxicity, iron and oxygen radicals, and chelation therapy. PMID- 8409696 TI - HTLV-1 and multiple sclerosis: the link is missing. PMID- 8409697 TI - ATP depletion and cell injury: what is the relationship? PMID- 8409698 TI - Alcohol sensitivity, alcohol metabolism, risk of alcoholism, and the role of alcohol and aldehyde dehydrogenase genotypes. PMID- 8409699 TI - Steroid hormone receptors. AB - Tremendous advances have been made in understanding the fundamental mechanisms of steroid hormone action. The cloning of the genes that encode classic steroid hormone receptors revealed that these receptors are but part of a large family of ligand-responsive transcriptional regulators. This fundamental advance has already contributed greatly to our understanding of a variety of diseases, particularly cancer, and the total dividend has not yet been realized. Steroid receptors have been and will be a fruitful area of research for many years. PMID- 8409700 TI - Anthracycline toxicity is potentiated by iron and inhibited by deferoxamine: studies in rat heart cells in culture. AB - The interrelation between iron, iron chelation, and anthracycline toxicity was investigated in a heart cell culture system. Two indicators of cellular damage have been used, lactate dehydrogenase (LDH) release and cell contractility. Both of these indicators have shown a marked increase in doxorubicin toxicity by prior iron loading. This was not a simple additive effect, because at the concentrations used, iron had only a minimal effect on LDH release and no effect at all on contractility, whereas doxorubicin had only a minor effect on contractility. Deferoxamine (DF) treatment of iron-loaded heart cells resulted in a marked decrease in anthracycline toxicity as judged both by LDH leakage and cell contractility. However, DF treatment of normal heart cells had no measurable protective effect against doxorubicin toxicity, whether DF was administered before or simultaneously with doxorubicin. Doxorubicin treatment did not alter cellular malondialdehyde (MDA) concentrations in either normal or iron-loaded cells. Conversely, the protective effect of DF in iron-loaded cells and its failure to prevent anthracycline toxicity in normal cells were both associated with a significant decrease in MDA measurements. Our data indicate that iron overload aggravates anthracycline toxicity and that this interaction may be prevented by effective iron chelating therapy. Because patients requiring anthracycline therapy often have increased tissue iron stores caused by multiple blood transfusions and bone marrow suppression, our observations may have important implications for the prevention of anthracycline toxicity. PMID- 8409701 TI - Presence of cross-reactive antibodies to HTLV-1 and absence of antigens in patients with multiple sclerosis. AB - Antibodies to HTLV-1, as determined by ELISA, were highly elevated in the serum samples of four out of four (100%) patients with TSP, moderately elevated in four out of four (100%) HTLV-1 carriers, slightly elevated in 12 out of 34 (35%) patients with MS, and absent from the serum samples of 34 normal subjects. Western blot analysis showed that the antibodies to HTLV-1 antigens in MS serum were heterogeneous. Cultivation of peripheral blood lymphocytes (PBLs) from patients with MS or normal subjects did not generate HTLV-1 core p19 antigen in the supernatant of culture medium, whereas cultivation of PBLs from patients with TSP and carriers of HTLV-1 generated core p19 antigen after 3 days for up to 28 days of cultivation. HTLV-1 antigens were also expressed on the surface of PBLs in three out of four patients with TSP and in two out of four HTLV-1 carriers on days 14 and 28 of cultivation, as measured by indirect immunofluorescence or alkaline phosphatase staining, but were not found in PBLs of any of 34 patients with MS or 34 normal subjects. The data indicate that although cross-reacting antibodies appear in the serum of some patients with MS, not enough evidence exists to suggest that HTLV-1 antigen is being produced in MS or that HTLV-1 plays a role in the pathogenesis of this disease. PMID- 8409702 TI - Effect of chemical hypoxia on intracellular ATP and cytosolic Mg2+ levels. AB - Intracellular magnesium is intimately associated with adenosine triphosphate (ATP) concentrations and energy utilization. We determined adenine nucleotide concentrations (ATP, adenosine diphosphate, and adenosine triphosphate) and the associated changes in intracellular free Mg2+ ([Mg2+]i) by high-performance liquid chromatography and fluorescent methods, respectively. Various mitochondrial inhibitors were used to deplete intracellular ATP and alter energy charge in epithelial cells. The opossum kidney (OK) cell line was used as a prototypic renal epithelial cell. These agents markedly deplete intracellular ATP levels with modest changes in [Mg2+]i and [Ca2+]i. Because these agents have disparate actions, it is likely that these changes were due to alterations in ATP rather than to selective drug effects. Cyanide resulted in a rapid (within 2 minutes) fall in ATP from 25.85 to 10.58 nmol/mg protein or about 3 mmol/L, whereas [Mg2+]i increased gradually (10 minutes), from 513 +/- 7 to 1096 +/- 105 mumol/L [Ca2+]i increased from 109 +/- 12 to 153 +/- 10 nmol/L within 20 seconds, then returned to basal concentrations. The changes in ATP, Mg2+, and Ca2+ were not altered by removing external Na+o, adding ruthenium red, or treating with vanadate. Antimycin diminished ATP levels in a manner similar to the effect of cyanide, but by contrast [Mg2+]i decreased to 436 +/- 13 mumol/L and [Ca2+]i transiently increased. These studies indicate that we are able to distinguish Mg2+ movements from those of Ca2+ by fluorescent techniques and suggest that intracellular regulation of [Mg2+]i is distinctive from that of [Ca2+]i. Oligomycin resulted in marked and rapid falls in [ATP]i with disproportionate increases in [Mg2+]i. The response of magnesium-depleted cells (basal [Mg2+]i, 231 +/- 10 mumol/L) after inhibitor-induced energy depletion was similar to that of control cells. These studies suggest that large changes in intracellular ATP levels do not markedly alter intracellular [Mg2+]i control and, in turn, that intracellular free Mg2+ is not a limiting factor in ATP metabolism after energy depletion with chemical hypoxia. PMID- 8409703 TI - Morphometric analysis of renal arterioles in subtotally nephrectomized rats. AB - Renal arteriolar diameters were measured by using vascular casts in the remnant kidneys of subtotally nephrectomized rats. Two weeks after nephrectomy, both afferent and efferent arterioles were dilated, with development of glomerular hypertrophy. Thirteen weeks after nephrectomy, the afferent arteriole was dilated further, while the efferent arterioles became constricted. Glomerular hypertrophy was augmented. At this stage, the systemic pressure was elevated, with development of marked glomerular sclerosis. Throughout the experiment, captopril lowered the systemic pressure, suppressed glomerular hypertrophy, and limited glomerular damage. Both the afferent and efferent arterioles were dilated further. Changes in the arteriolar diameters and the systemic blood pressure suggested that elevated glomerular pressure existed in the remnant kidney and did not exist in the captopril-treated remnant kidney throughout the experiment. Hydralazine and trichloromethiazide therapy lowered the systemic pressure and maintained the arteriolar diameters and glomerular size at normal levels. However, at the late stage, afferent and efferent arterioles were dilated, with development of glomerular hypertrophy and severe glomerular sclerosis. The results suggested that elevated glomerular pressure was involved in development of glomerular sclerosis. However, factors other than hemodynamics should be considered in the pathogenesis of glomerular sclerosis. A direct causal relationship between glomerular hypertrophy and glomerular damage was not shown because captopril ultimately limited glomerular sclerosis despite glomerular hypertrophy at the early stage, and hydralazine and trichloromethiazide therapy did not ultimately ameliorate the glomerular sclerosis despite normal glomerular size at the early stage. PMID- 8409704 TI - Genetic predisposition to hypertension and microvascular injury in the remnant kidney model. AB - Genetic factors have been implicated in the development and progression of glomerulosclerosis and nephron loss in both experimental animals and in humans. The influence of a differing genetic predisposition to hypertension was examined in the remnant kidney (RK) model of progressive glomerulosclerosis. Dahl salt sensitive (S) and salt-resistant (R) rats fed a normal salt diet underwent either sham surgery or approximately 5/6 renal ablation and were studied 2 to 3 weeks later. Renal ablation resulted in significantly more severe hypertension in RK-S rats (205 +/- 6.3 mm Hg, mean +/- SEM) compared with RK-R rats (153 +/- 3.5 mm Hg; p < 0.01). Renal autoregulatory ability, a protective mechanism against renal transmission of systemic hypertension, was normal in both S and R rats with intact renal mass. Renal ablation resulted in similar impairments of renal autoregulatory ability in both strains. However, striking differences in the severity of renal microvascular and glomerular injury were observed between the remnant kidneys of S and R rats, paralleling the differences in the severity of hypertension. The RK-S rats exhibited acute fibrinoid necrosis and thrombosis of glomerular capillaries, arterioles, and small arteries, whereas only mild segmental glomerulosclerosis lesions were observed in a small percentage of glomeruli in the RK-R rats. The intact kidneys of both strains were essentially free of glomerular or vascular lesions. These data suggest that a genetic predisposition to hypertension is a major determinant of the severity of hypertension that follows severe reduction of renal mass and the severity of the resulting hypertension, in turn, critically influences the severity of glomerular injury in the RK model. PMID- 8409705 TI - Inflammatory responses to implanted polymeric biomaterials: role of surface adsorbed immunoglobulin G. AB - In many cases, evidently inert and nontoxic biomaterials may trigger procoagulant and inflammatory responses. Because most polymeric biomaterials accumulate a surface layer of protein immediately after implantation, these adverse reactions may stem from secondary interactions between the host and this surface layer of adsorbed proteins. Using polyester terephthalate (the polymer from which both Dacron and Mylar are produced) as a model, we have explored the hypothesis that surface-adsorbed immunoglobulin might mediate subsequent inflammatory responses. We find, as have others, that immunoglobulin G (IgG) does spontaneously adsorb to polymer surfaces, both in vitro and in vivo. Furthermore, polymer implants precoated with IgG do activate human polymorphonuclear neutrophils in vitro and also attract substantial numbers of phagocytes (especially polymorphonuclear neutrophils and macrophages) when implanted in mice. However, when implants are placed in mice having a form of severe combined immunodeficiency (and, consequently, almost undetectable levels of serum IgG), a near-normal influx of phagocytic cells ensues. Thus, spontaneously-adsorbed surface IgG does not appear to be a necessary precedent to inflammatory responses directed against implanted biomaterials. PMID- 8409706 TI - A role for endothelin and nitric oxide in the pressor response to diaspirin cross linked hemoglobin. AB - Diaspirin-cross-linked hemoglobin (DCLHb; Baxter Healthcare Corp) has potential for clinical use as a hemoglobin-based oxygen-carrying solution. We have previously shown that DCLHb administration is associated with a self-limiting increase in mean arterial pressure (MAP). Based on in vitro studies with other hemoglobin solutions, this vasopressor effect is thought to be mediated at least in part by the release or inhibition (or both) of endothelium-derived vasoactive substances. The purpose of our studies was to determine the role of endothelin (ET), a potent vasoconstrictor peptide, and nitric oxide, a vasodilator, in mediating the pressor effect of DCLHb in conscious rats. Intravenous administration of DCLHb has been shown to elicit an immediate increase in MAP that peaks within 30 minutes and returns to baseline by 300 minutes in a dose dependent fashion. Phosphoramidon, an inhibitor of proendothelin conversion to ET, attenuated the elevation of MAP when administered before DCLHb. Administration of cyanomet DCLHb, a DCLHb molecule that is unable to interact with NO, was not associated with an elevation of MAP. L-arginine, the substrate for NO synthesis, and nitroglycerin, an NO donor, significantly reduced MAP when infused 15 minutes after DCLHb administration. Based on these findings, we conclude that the DCLHb-induced elevation of MAP in vivo is mediated at least in part by ET and the inhibition of NO. Although these data support earlier reports of hemoglobin binding NO, this is the first report of the pressor response to hemoglobin being attenuated by an agent that blocks the conversion of proendothelin to ET. PMID- 8409707 TI - Apoptosis and pleomorphic micromitochondriosis in the sinus nodes surgically excised from five patients with the long QT syndrome. AB - Sinus nodes of five symptomatic patients with the long QT syndrome were surgically excised and followed by permanent electronic pacing as part of a new surgical treatment. We examined those sinus nodes by light and electron microscopy with tissue that was promptly fixed at the time of surgery. All five sinus nodes were similarly abnormal. By light microscopy we found distinctive focal fibrosis, some degenerating myocytes and neural elements, and numerous narrowed small vessels. Except in the nerves there was no evidence of inflammation. In electron micrographs the mitochondria within nodal myocytes were abnormally abundant, remarkably pleomorphic, and smaller than those in normal human sinus nodal cells. The ultrastructural features of the degenerated nodal cells were typical of apoptosis, characterized by the absence of inflammation, well-preserved mitochondria, the presence of apoptotic bodies, phagocytosis of these cells by neighboring myocytes, and especially in smooth muscle cells of arterioles, nuclear chromatin margination and nucleolar disintegration. Apoptotic degeneration of nodal myocytes was stochastic, with adjacent cells appearing unaffected. Focal ischemia caused by narrowed vessels may be a contributory factor, and the nerves may harbor some viral infection, but for the nodal myocytes the abnormality appears to be primarily apoptosis, sometimes called programmed cell death. Both the typically episodic clinical features and the terminal event in fatal cases of the long QT syndrome may be due to apoptosis. PMID- 8409708 TI - Evidence that the arg1744 gly1745 asp1746 sequence in the GPIIb-IIIa-binding domain of von Willebrand factor is involved in platelet adhesion and thrombus formation on subendothelium. AB - Previous studies have strongly suggested that the initial attachment (adhesion) of platelets to subendothelium or collagen at high shear rates is mediated by the binding of von Willebrand factor (vWf) to platelet glycoprotein Ib (GPIb). The finding in the present study that incubating human umbilical artery subendothelium with a monoclonal antibody to the GPIb binding site of vWf markedly inhibited platelet adhesion is fully consistent with this hypothesis. Platelet adhesion (and thrombus formation) also requires GPIIb-IIIa, and previous studies have suggested that the binding of vWf to GPIIb-IIIa may be involved in this process. To explore this further, we utilized two monoclonal antibodies (152B-20 and 152B-6) that recognize the arg1744 gly1745 asp1746 (RGD) sequence of vWf. These antibodies selectively inhibit vWf binding to GPIIb-IIIa and have no cross-reactivity with other RGD-containing proteins that can bind to GPIIb-IIIa. When added to citrated human blood, these antibodies (152B-20 in particular) inhibited platelet adhesion and thrombus formation on rabbit subendothelium at a shear rate of 2600 sec-1 but not at 400 sec-1. The results of the study thus provide direct evidence that the binding of vWf to GPIIb-IIIa is important for platelet adhesion and thrombus formation on subendothelium at high shear rates and that the arg1744 gly1745 asp1746 sequence in the mature vWf is involved in this process. PMID- 8409709 TI - Effects of alpha 1-proteinase inhibitor on chemotaxis and chemokinesis of polymorphonuclear leukocytes: its possible role in regulating polymorphonuclear leukocyte recruitment in human subjects. AB - A variety of biologic products derived from bacteria, inflammatory cells, and active degraded proteins have been identified as having chemotactic activity essential for polymorphonuclear leukocyte (PMN) recruitment to the site of inflammation. Little is known, however, concerning factors responsible for regulating the intensity and duration of PMN recruitment. Evidence is growing that proteinase inhibitors modify the migrating ability of PMNs, although the physiologic implications of this have eluded clarification. In an attempt to hypothesize a role of alpha 1-proteinase inhibitor (alpha 1-Pi) in PMN recruitment to inflammatory sites, we examined the effects of alpha 1-Pi in different physiologic concentrations on PMN migration with a microchemotaxis chamber technique. Alpha 1-proteinase inhibitor had both stimulatory and inhibitory effects on cell migration, depending on its concentration. The inhibitor was active in inducing both directed locomotion (chemotaxis) and nondirected locomotion (chemokinesis) in concentrations of 0.02, 0.2, and 2 mg/ml, with maximum potency in both cases at 0.2 mg/ml (corresponding to the normal alveolar surface fluid level in the lung). Alpha 1-proteinase inhibitor impaired chemotactic responsiveness to known chemoattractants at 2 and 10 mg/ml (corresponding to normal and inflammatory blood levels, respectively) in order of potency. These results suggest that alpha 1-Pi may play a role in regulating inflammatory processes, especially in the lung, through its stimulatory and inhibitory effects, depending on its concentration. PMID- 8409710 TI - Heme oxygenase is not expressed as a stress protein after renal ischemia. AB - Stressful stimuli such as heat, oxidative stress, heavy metals, and tissue trauma induce the expression of a family of proteins commonly referred to as stress proteins or heat shock proteins. The functions of these proteins are varied but include glycolysis, antioxidant defense, and several postulated "chaperone" functions involving the folding, unfolding, and translocation of other proteins. Heme oxygenase, the enzyme that catalyzes the degradation of heme to biliverdin, is also heat inducible and is, therefore, a heat shock protein. In the kidney, ischemia has been observed by several investigators to induce expression of the more commonly studied heat shock proteins HSP 70 and HSP 72. In addition, exposure of the kidney to myoglobin after glycerol injection induced heme oxygenase. The purpose of this study was to determine whether heme oxygenase is expressed as a stress protein after renal ischemia. Renal ischemia was induced in rats after right nephrectomy by clamping the renal artery for 40 minutes. Gene expression was evaluated after 60 minutes to 96 hours of postischemic reperfusion. There was essentially no expression of heme oxygenase at any of the time points evaluated. The absence of heme oxygenase expression was in striking contrast to the prompt and dramatic expression of HSP 70. This finding is consistent with the concept that all "stress proteins" are not equivalent and that, although there is considerable overlap between heat-sensitive gene promoters and oxidant stress-sensitive gene promoters, there is specificity for the type of stimulus that is able to activate any given stress protein gene. PMID- 8409711 TI - We will survive. PMID- 8409712 TI - Bronchopulmonary disease: an association with ulcerative colitis and pyoderma gangrenosum. PMID- 8409713 TI - A mnemonic approach to health reform. PMID- 8409714 TI - Profound deafness treated by the Ineraid multichannel intracochlear implant. AB - Twelve deaf adults and two deaf children were treated with the Ineraid (formerly Symbion) four channel intracochlear implant between September 1989 and October 1991 at Addenbrooke's Hospital in Cambridge. All were post-lingually totally deaf and had found themselves beyond the reach of hearing aids. The effect of the implant upon the patients ability to lip-read was tested with the speech tracking test, BKB sentences (comparable to CID sentences) and Boothroyd word lists (comparable to NU6 word lists). All patients showed an improvement in their ability to understand speech with the help of the implant. Discrimination of speech without lip-reading was tested with Boothroyd word lists and BKB sentences, eight patients (57 per cent) demonstrated some 'open set' speech discrimination. The acceptability of the carbon percutaneous pedestal is discussed from the patient's, audiologist's and surgeon's points of view. PMID- 8409715 TI - Gentamicin ototoxicity in continuous ambulatory peritoneal dialysis. AB - A prospective study was undertaken of 10 chronic renal failure patients on Continuous Ambulatory Peritoneal Dialysis (CAPD) complicated by repeated bouts of peritonitis treated with gentamicin. Each 10-day treatment course consisted of a 120 mg loading dose, followed by 16 mg in 21 of peritoneal dialysate, given four times a day. Serum gentamicin analysed by enzyme immunoassay showed a mean level of 5.2 micrograms/ml, (range 3.7 to 6.6 mg/ml) four hours after the loading dose. Similar levels, well within the therapeutic range, were maintained on the 3rd, 5th, 7th and 9th days of intraperitoneal gentamicin therapy, suggesting no accumulation of gentamicin in the serum. Pure tone audiometry, electronystagmography and clinical assessment were performed during each course of treatment. Although no evidence of ototoxicity was found during the first two courses of gentamicin, but disequilibrium and bobbing oscillopsia were present during the third and fourth courses of gentamicin. These findings could be explained by cumulative injury to the vestibular apparatus caused by repeated therapeutic insults. PMID- 8409716 TI - The use of cortical bone grafts in ossiculoplasty. I: Surgical techniques and hearing results. AB - Concern about the possible risk of viral infection being transmitted by the use of homografts has renewed interest in cortical bone autografts as an ossicular substitute. Over the last four years I have used cortical bone on a regular basis. Comparison of hearing results obtained using ossicular and cortical bone grafts shows no significant difference between the two groups. Cortical bone appears to be satisfactory material for ossiculoplasty, but long-term studies of outcome are required to confirm this. PMID- 8409717 TI - The use of cortical bone grafts in ossiculoplasty, II: Graft mass and hearing change at different frequencies. AB - The masses of preserved ossicles and cortical bone grafts have been studied in vitro using a Stanton Unimatic CL41 balance. The cortical bone grafts prepared for use in malleus-stapes assemblies had a mean mass 28 per cent greater than that for an incus, while those prepared for use in a malleus-footplate or drum footplate assembly had a mean mass 58 per cent greater. Analysis of the hearing results for the first 20 ossiculoplasty operations performed by the author using cortical bone grafts revealed no significant differences in outcome attributable to increased mass. However, large grafts did become fixed to surrounding structures in some cases. PMID- 8409718 TI - The use of cortical bone grafts in ossiculoplasty. III: Audit of costs. AB - This paper describes the results of a comparative audit of the costs of using artificial prostheses and cortical bone. The costs of using a bone graft have been calculated by estimating the expenses incurred as a result of the additional time required for preparation of the grafts. The mean cost of preparing a graft was 29.10 pounds, while the mean cost of using a prosthesis was 65.01 pounds. PMID- 8409719 TI - Lateral temporal bone resection for extensive pinnal malignancy. Has anything changed in forty years? AB - This paper offers an account of the contemporary surgical approach to advanced tumours of the external ear based on a series of 11 patients. There were eight squamous, two basal cell carcinomas and one mucoepidermoid tumour. The traditional method of excision was slightly modified by performing microsurgical dissection of the lateral part of the temporal bone rather than chisel osteotomies, and then including it en bloc with the involved soft tissues. The defect was then closed using a scalp or myocutaneous flap and this combination of otological and reconstructive expertise has proved satisfactory. Four patients are alive with no evidence of disease a mean of 4.2 (range 1.0-7.0) years from surgery: two patients who remained free of disease have subsequently died of unrelated conditions 12 and 24 months post-operatively, and in three cases death from recurrent disease occurred a mean of 1.4 (range 0.9-2.1) years after our surgery. There were two post-operative deaths. Based on the actuarial survival of 36 per cent and a successful disease clearance rate of 54 per cent, our conclusion is that the outlook of this condition has not dramatically improved since the original descriptions of the management of this problem first appeared, although intervention remains justifiable because of the potential curability and relief of symptoms. PMID- 8409720 TI - Day-case tonsillectomy: a review of the literature. AB - Adenotonsillectomies are performed increasingly as day cases in North America and Canada. This paper analyses reports where such tonsillectomies were performed. It is possible to successfully select a group with lower risk of complications. Haemorrhage occurs either early in the post-operative period or later than 24 hours after operation. With good organization tonsillectomies can be safely performed as day cases. PMID- 8409721 TI - Same-day tonsillectomy. AB - Tonsillectomies have been performed on a same-day basis in Coventry for the past three years. We report our experience with this technique which has gradually evolved over the last ten years. The procedure is described in detail and the results of the first two years have been analysed. In common with the experience of others, it is possible to perform tonsillectomies safely on a same-day basis. PMID- 8409722 TI - Parental preferences for duration of hospital stay following tonsillectomy. AB - Parents of children undergoing (adeno) tonsillectomy were questioned about their preferences for a period of hospitalization. The proportion wanting early discharge increased as their children recovered after surgery. PMID- 8409723 TI - A study of haemostasis following tonsillectomy comparing ligatures with diathermy. AB - The use of diathermy to achieve haemostasis after tonsillectomy remains controversial. We have reviewed the English language literature, and found no convicting evidence that diathermy is any more likely to cause post-operative haemorrhage than the use of ligatures. The results of a prospective, randomized study of 1036 consecutive tonsillectomies are presented. No significant difference was found in post-operative haemorrhage rates when either diathermy or ligatures were used. Diathermy was found to reduce operating time compared to ligatures. The possibilities for day-case tonsillectomy are discussed. PMID- 8409724 TI - Rhabdomyosarcoma of the adult head and neck: a clinicopathological and DNA ploidy study. AB - We investigated the DNA ploidy patterns in thirteen primary and four recurrent rhabdomyosarcomas of the head and neck from thirteen adult patients and correlated the findings with other clinicopathological factors and clinical outcome. Twelve (92.7 per cent) of the primary neoplasms manifested an aneuploid DNA pattern, five had more than one stemline, and one neoplasm displayed a diploid DNA pattern. All recurrent lesions were DNA aneuploid with DNA indices (DIs) corresponding to their primary neoplasms. No correlation between the ploidy pattern and histological subtypes, tumour location, clinical stage and patient's clinical course was found. In this study, only two patients were long-term survivors. Both patients had stage I neoplasms that were located in non parameningeal sites which manifested an alveolar histological pattern. Our data indicate that adult rhabdomyosarcomas of head and neck are preponderantly DNA aneuploid and are highly aggressive malignant neoplasms. Our results also suggest that tumours which are low stage and in a non-parameningeal location may pursue a less aggressive course. PMID- 8409725 TI - Temporal bone dissection using a low cost miniature electric drill. AB - The performance of various specialized and general purpose drills was assessed for use in temporal bone dissection. The Minicraft MB 120 and MB 130 were found to be as effective as specialized drills but with greater convenience and much reduced cost. The use of these drills allows high quality temporal bone dissection to be performed on a limited budget. PMID- 8409726 TI - Autosuture GIA gun: a new application in the treatment of hypopharyngeal diverticula. AB - A new surgical technique for the treatment of moderate to large hypopharyngeal diverticula is described. The procedure is particularly useful in debilitated patients in that it is quick and guarantees closure of the party wall between the pouch and upper oesophagus. PMID- 8409727 TI - Severe cochlear dysplasia causing recurrent meningitis: a surgical lesson. AB - Meningitis may be the sole presenting sign of a cerebrospinal fluid (CSF) fistula of the temporal bone. An eight-year-old boy suffering from recurrent meningitis was found to have bilateral severe cochlear dysplasia. Bilateral tympanotomies were performed, planning to obliterate each vestibule. In the right ear a stapedectomy was performed, resulting in a torrential 'CSF gusher' and difficulty in packing the vestibule. CSF rhinorrhoea requiring revision surgery and two episodes of gram-negative bacterial meningitis complicated the post-operative management, resulting in a prolonged hospital stay. Subsequently, the left ear was managed in a different fashion, leaving the stapes in situ, with grafts placed to seal the oval window niche. We would recommend this alternative procedure in cases of severe cochlear dysplasia, where abnormalities of the vestibule and basal turn of the cochlea mean that performing a stapedectomy to pack the vestibule may result in a severe 'CSF gusher', by opening directly into the subarachnoid space. PMID- 8409728 TI - Post-traumatic bilateral facial palsy. AB - Bilateral facial palsy is an uncommon condition and few clinicians have experience of more than the occasional case. Unlike its unilateral counterpart, an aetiological factor is often demonstrable. A wide variety of recognized diseases may give rise to the condition and thus thorough investigation is required. This paper presents an unusual example of this rare condition. PMID- 8409729 TI - Oncocytic mixed nasal tumour. PMID- 8409730 TI - A wooden orbital foreign body. AB - A case is reported in which a patient presented, 18 months after initial injury, with a progressive proptosis of the left eye. X-rays did not reveal a foreign body but a fracture of the lateral orbital wall and an area of soft tissue density were seen on the CT scan. Exploration of the orbit revealed a 2 cm wooden fragment in the floor of the orbit. PMID- 8409731 TI - Osteoblastoma of the nasal cavity. AB - The clinicopathological features of a rare case of osteoblastoma of the nasal cavity arising from the nasal turbinate are reported and compared with four reported cases of osteoblastoma with nasal cavity involvement. Two of the five tumours involved the nasal cavity and paranasal sinuses. The remaining three tumours were confined in the nasal cavity; one arose from nasal bone and two from nasal turbinate periosteum. Four tumours were successfully treated with local excision. One tumour recurred locally after excision; the recurrence was apparently controlled by further local excision. PMID- 8409732 TI - Leiomyoma of the paranasal sinuses. AB - The paranasal sinuses are a rare site for tumours of myogenic origin. There has been only one previously reported case in the English literature. We present a case of a leiomyoma filling the anterior ethmoid sinus and middle meatus which was excised via a Patterson's external ethmoidectomy. PMID- 8409733 TI - Synovial osteochondromatosis of the temporo-mandibular joint. AB - A rare case of synovial osteochondromatosis of the temporo-mandibular joint is presented. Important diagnostic information can be obtained by CT, MRI, 99m Tc bone scan and aspiration biopsy. Observation using a light microscope showed mild cellular atypia, but the hallmarks characteristic of chondrosarcoma were not found. Observation using an electron microscope showed the mature chondrocytes contained a well-developed rough endoplasmic reticulum. Histological evaluation indicated that the present case was benign synovial osteochondromatosis in an early stage. PMID- 8409734 TI - Palato-pharyngo-laryngeal myoclonus: an unusual cause of dysphagia and dysarthria. AB - We describe a case of palato-pharyngo-laryngeal myoclonus, an unusual variant of palatal myoclonus, which presented with dysphagia and dysarthria. The aetiology and presenting features of myoclonus are discussed. Various treatment options are considered. PMID- 8409735 TI - Pectoralis major muscle transposition: an adjunct to laryngeal preservation in severe chondroradionecrosis. AB - Radiation-induced chondronecrosis of the larynx in early glottic cancer is a rare but serious complication, sometimes requiring total laryngectomy. We describe two patients who developed the major complication of chondroradionecrosis of the larynx 11 and 18 years after completion of radiotherapy. Both cases underwent excision of the overlying skin and of all infected and devitalized thyroid cartilage, followed by pectoralis major muscle transposition and split thickness skin grafting. This procedure resulted in a preservation of all laryngeal functions and also an acceptable cosmetic outcome. PMID- 8409736 TI - Upper airway invasion by well-differentiated thyroid carcinoma. AB - Invasion of the upper airway by well-differentiated thyroid carcinoma is very unusual. Treatment is primarily surgical resection, but the extent of the resection has been controversial. Adjuvant treatment using I131 or radiotherapy also has an important role in management. We describe two cases of direct infiltration of the larynx and trachea by well-differentiated thyroid carcinoma and discuss the current recommendations for the management of this difficult problem. PMID- 8409737 TI - Frozen section diagnosis in a jugulo-tympanic paraganglioma. AB - The light microscopy and immunohistochemical findings of a jugulo-tympanic paraganglioma occurring in a 29-year-old man are reported. Diagnostic difficulties from frozen sections are stressed and selected literature is briefly commented upon. PMID- 8409738 TI - Day-case tonsillectomy--a public demand? PMID- 8409739 TI - Pediatric chronic illness. PMID- 8409740 TI - Pediatric chronic illness. PMID- 8409741 TI - Integrative strategy instruction: an elusive ideal for teaching adolescents with learning disabilities. PMID- 8409742 TI - Student-centered or strategy-centered instruction: what is our purpose? PMID- 8409743 TI - The promise and pitfalls of integrated strategy instruction. PMID- 8409744 TI - Comments on Ellis's integrative strategy instruction model. PMID- 8409745 TI - Teaching strategy sameness using integrated formats. PMID- 8409746 TI - Fostering critical thinking skills in students with learning disabilities: an instructional program. AB - An instructional unit was developed to enhance the critical thinking skills of middle school and high school students with learning disabilities by teaching them the principles of scientific reasoning. Student-teacher dialogues were used to engage the students in an active process of critical inquiry. Students analyzed everyday information presented in brief magazine articles and advertisements by applying the principles of the scientific method. The students made statistically significant improvements in their ability to (a) identify the principal claim made in an article or advertisement, (b) graph the relevant data, and (c) evaluate the claims made in the article and explain their support or rejection of the claims based on data. After the instructional unit, the overall performance of the special education students exceeded that of a control group composed of regular education students who had not received instruction in critical thinking. PMID- 8409747 TI - Chemical dependency in students with and without learning disabilities. AB - To determine if students with learning disabilities (LD) demonstrate a higher frequency of chemical dependency than students without learning disabilities (NLD), a total of 191 adolescents with LD (101 males and 90 females) were given the Substance Abuse Subtle Screening Inventory (SASSI). The sample consisted of 88 students with LD and 103 NLD students between the ages of 12 and 18. The SASSI is an objectively scored self-report inventory that accurately classifies adolescents as chemically dependent (CD) or not chemically dependent (NCD). A significantly higher proportion of students with LD than NLD students were classified as CD. Of the 30 students who were classified as CD, 70% were students with LD. A discriminant analysis indicated that the presence or absence of a learning disability was a better predictor of classification as CD or NCD than gender, ethnicity, age, socioeconomic status, or family composition. The implications for evaluation and educational planning for students with learning disabilities are discussed. PMID- 8409748 TI - Inhibition of human mononuclear cell proliferation, interleukin synthesis, mRNA for IL-2, IL-6, and leukotriene B4 synthesis by a lipoxygenase inhibitor. AB - Human peripheral blood mononuclear cells (PBMCs) were cultured in vitro with various stimuli and in the presence or absence of a 5-lipoxygenase (5-LO) inhibitor, A-63162, to measure its effects on PBMC proliferation, interleukin-2 receptor (IL-2R) expression, interleukin-2 (IL-2) synthesis, interleukin-6 (IL-6) synthesis, and accumulation of messenger RNA for IL-2 or IL-6. A-63162 inhibited PBMC proliferation stimulated by phytohemagglutinin (PHA) and phorbol myristate acetate (PMA) plus A23187, IL-2 receptor expression stimulated by PHA, and IL-2 or IL-6 synthesis induced by PHA plus PMA or PMA plus A23187. At the same concentration, A-63162 inhibited accumulation of mRNA for IL-2 or IL-6 and also inhibited leukotriene B4 (LTB4) synthesis. Our data indicate that the 5-LO inhibitor A-63162 has immunosuppressive activity that may be due to inhibition of LTB4 production or to direct inhibitory actions of A-63162 on IL-2 and IL-6 synthesis. PMID- 8409749 TI - Granulocytes enhance LPS-induced tissue factor activity in monocytes via an interaction with platelets. AB - In the present study we have investigated the effect of platelets and granulocytes on bacterial lipopolysaccharide (LPS)-induced tissue factor (TF) activity in monocytes. Experiments were performed on freshly isolated cells resuspended in heparinized plasma and recombined with platelet-poor or platelet rich plasma. In a platelet-dependent reaction the granulocytes enhanced LPS induced TF activity by an average of 100%. The effect was dose dependent with regard to the number of both granulocytes and platelets, respectively. Granulocytes and/or platelets did not affect LPS-induced tumor necrosis factor (TNF) secretion from monocytes. Phorbol myristate acetate (PMA) per se was not able to induce TF activity in our system. In contrast, the agonist caused a substantial increase in TF activity induced by LPS. The effect was totally dependent on the presence of platelets and was shown to be due to stimulation of both granulocytes and monocytes (the activity rose from 30 +/- 7 to 83 +/- 12 mU/10(6) cells in the presence of platelets and from 69 +/- 8 to 143 +/- 22 mU/10(6) cells in the presence of platelets and granulocytes). Effects similar to those observed with PMA were obtained with physiological concentrations (10 ng/ml) of TNF. A combination of these two agonists gave no further amplification of LPS-induced TF activity compared with the effect of the agonists separately. Low concentrations of a monoclonal anti-CD15 antibody abolished the stimulatory effects of platelets and granulocytes. Furthermore, the anti-CD15 antibody neutralized the effect of TNF, whereas the PMA effect was reduced by almost 75%. These results were confirmed in a whole-blood system. The inhibitory effect of the antibody may be associated with CD15's role as a complementary ligand for PADGEM. Our study indicates that a close interaction between granulocytes, platelets, and monocytes is essential for optimal TF activity induced by LPS. It is hypothesized that the effect of granulocytes is related to their ability to activate platelets. We propose that upon activation granulocytes secrete a product that enhances the capacity of platelets to stimulate TF activity in monocytes. PMID- 8409750 TI - Inhibition of apoptosis and prolongation of neutrophil functional longevity by inflammatory mediators. AB - Neutrophil apoptosis leads to macrophage ingestion of intact senescent neutrophils. This may represent a neutrophil removal mechanism that is important both in the control of inflammatory tissue injury and for the normal resolution processes of inflammation. Because apoptosis is likely to be a key control process in cell and tissue homeostasis, a number of inflammatory mediators were tested for their ability to modulate the rate of apoptosis in populations of neutrophils aging in culture. Endotoxic lipopolysaccharide, human recombinant complement factor 5a, and human recombinant granulocyte-macrophage colony stimulating factor all markedly inhibited the rate of neutrophil apoptosis in a concentration-dependent fashion, without inducing necrosis (as assessed by trypan blue exclusion). This inhibitory effect on the rate of neutrophil apoptosis was shown by morphological criteria and confirmed by gel electrophoresis of extracted DNA. Inhibition of apoptosis of aging neutrophil populations was associated with prolongation of the functional life span of the population as assessed by the ability of neutrophils to spread on glass surfaces, to polarize in response to deliberate stimulation with N-formyl-Met-Leu-Phe (fMLP), and to release the granule enzyme marker myeloperoxidase on fMLP stimulation. These observations show that inflammatory mediators prolong the functional life span of neutrophils through modulation of apoptosis. Further elucidation of these mechanisms will lead to a better understanding of the processes controlling neutrophil residence and function in inflamed tissues and may provide further insights into the molecular mechanisms of apoptosis, which is of widespread importance in tissue biology. PMID- 8409751 TI - Autocrine amplification of PAF-acether formation in immunologically activated murine macrophages. AB - When murine macrophages activated in vivo with bacille Calmette-Guerin were triggered with either acetyl-CoA or propionyl-CoA to form PAF-acether (PAF), similar amounts of platelet-aggregating product were recovered. Liquid chromatographic purification and reversed-phase analysis showed that the composition of PAF molecular species formed in the presence of acetyl-CoA was an equimolar mixture of PAF bearing C16:0 alkyl chain (57% +/- 7, mean +/- SD, n = 3) and PAF C18:1. The PAF-like material obtained from the propionyl-CoA supplemented macrophages was a mixture of the propionyl analogue of PAF (66% +/- 11, n = 3) and native PAF. The rate of lyso-PAF:acetyl-CoA acetyltransferase (EC 2.3.1.67) reaction in a macrophage lysate was similar for either substrate in the presence of an equimolar mixture of propionyl-CoA and acetyl-CoA. We conclude that the exogenously added propionyl-CoA is transferred to lyso-PAF acceptor to form propionyl-PAF by the PAF-forming acetyltransferase. Propionyl-PAF triggers the formation of native PAF probably from the endogenous acetyl-CoA pool. Two specific PAF antagonists, BN 52021 (60 microM) and WEB 2086 (3 microM), did not influence the rate of PAF synthesis in the presence of either acetyl-CoA or propionyl-CoA and did not prevent native PAF formation when propionyl-CoA was added alone, suggesting that the classical PAF receptors are not involved. This is the first description of a possible mechanism of autocrine amplification of PAF biosynthesis in macrophages. PMID- 8409752 TI - Luminol-enhanced chemiluminescence induced in peripheral blood-derived human phagocytes: obligatory requirement of myeloperoxidase exocytosis by monocytes. AB - Luminol-enhanced chemiluminescence (LCL) of phagocytes is often used to monitor for the generation of reactive oxygen mediators. A strong LCL reaction, as observed in triggered peripheral blood monocytes and neutrophils, depends on both the activation of an NAD(P)H-dependent oxidase and a functional myeloperoxidase (MPO). The aim of this work was to compare the LCL response induced by soluble and particulate stimuli in monocytes with that of polymorphonuclear leukocytes (PMNs). In agreement with earlier results, neutrophils showed a first peak of LCL activity within 1 min and a second peak around 3 min when stimulated with soluble stimuli. The first peak is sensitive to oxygen scavengers and requires the presence of extracellular luminol, whereas the second peak is much less sensitive to oxygen scavengers and does not require the presence of extracellular luminol. The first peak of LCL is therefore thought to represent extracellular and the second peak intracellular LCL. Monocytes, in contrast, showed only the first peak of LCL activity. This peak was sensitive to oxygen scavengers, required the extracellular presence of luminol, and could be suppressed by a rapid pH shift to a pH not allowing LCL (i.e., to pH 5). These results suggest that the stimulus dependent extracellular release of oxygen metabolites and of MPO is an obligatory requirement for LCL induced in monocytes. An exclusively extracellular LCL was noted upon stimulation with aggregated immunoglobulin G, f-Met-Leu-Phe, phorbol myristate acetate, A23187, anti-CD36 antibodies, opsonized zymosan, and opsonized E. coli. With the addition of H2O2 in excess (1 microM), the LCL response of monocytes therefore reflected the stimulus-dependent degranulation of MPO, and LCL-based determination of MPO release closely correlated with enzymatic MPO determination. The question of why LCL induced in monocytes, but not in PMNs, is restricted to the extracellular compartment was addressed. Although flow cytometric experiments were consistent with the hypothesis that extracellular H2O2 entered the cytoplasm of monocytes less efficiently than that of PMNs, other more important factors are assumed to contribute. Collectively, these results point to previously unrecognized differences in monocyte and PMN LCL which may reflect cellular differences of potential pathophysiological significance. PMID- 8409753 TI - The in vivo effects of rhIL-1 alpha therapy on human monocyte activity. AB - Pleiotropic cytokines such as interleukin-1 alpha (IL-1 alpha) have multiple effects on peripheral blood monocytes (PBMs). This study examined the ability of in vivo recombinant human IL-1 alpha (rhIL-1 alpha) therapy to enhance clinically important monocyte functions in ovarian cancer patients prior to chemotherapy. After 4 days of continuous infusion, in vivo rhIL-1 alpha therapy amplified both the number and activity of PBMs. Therapy with rhIL-1 alpha increased the number of PBMs sixfold. These monocytes had a significantly increased ability to produce superoxide anion in response to phorbol 12,13-dibutyrate stimulation. Their ability to secrete spontaneously the immunomodulatory cytokines IL-1 alpha and IL 1 beta was significantly increased, but their ability to secrete tumor necrosis factor alpha (TNF-alpha) was not significantly elevated. These effects of rhIL-1 alpha infusion on cytokine secretion by PBMs appear to be related to rhIL-1 alpha induced increases in the mRNA levels for these cytokines. In contrast, rhIL-1 alpha therapy did not significantly alter PBM response to lipopolysaccharide (10 micrograms/ml). In summary, infused rhIL-1 alpha, in addition to its use as a myeloprotective agent, has enhancing effects on the number and activity of PBMs. The effects of rhIL-1 alpha infusion on PBM function demonstrated here should at least transiently increase the ability of monocytes to combat infection and enhance host immune response. PMID- 8409754 TI - A single exogenous stimulus activates resident rat macrophages for nitric oxide production and tumor cytotoxicity. AB - Based on experiments with mouse cells, it appears that macrophage activation for cytotoxicity requires two exogenous signals. One signal primes or sensitizes the cells, while the second activates them for killing. The present studies were designed to analyze the capacity of rat macrophages to be activated for nitric oxide production and for cytotoxicity by different inflammatory stimuli. We found that both resident alveolar macrophages (AMs) and resident peritoneal macrophages (PMs) from Sprague-Dawley rats produced nitric oxide in response to relatively low doses of a single exogenous activating stimulus [interferon-gamma (IFN-gamma) or lipopolysaccharide (LPS)]. Resident PMs treated with either of these agents alone also exhibited nitric oxide-mediated cytotoxicity toward xenogeneic and allogeneic tumor targets. In contrast to results reported previously with both resident and elicited PMs from mice, tumor necrosis factor-alpha (TNF-alpha) exerted only a small enhancing effect on IFN-gamma-induced nitric oxide production by resident rat PMs. In addition, although some level of cooperativity between IFN-gamma and LPS was observed at low concentrations of LPS (< 10 ng/ml), IFN-gamma did not augment the effects of higher concentrations of LPS (> or = 10 ng/ml) on nitric oxide production by PMs. In contrast to PMs, AMs had a strong synergistic response to combinations of IFN-gamma and LPS or TNF-alpha but also only at relatively low concentrations of IFN-gamma and LPS. Furthermore, maximum nitric oxide production induced by IFN-gamma in these cells could be enhanced by TNF-alpha or low doses of LPS. However, as observed with PMs, combinations of IFN gamma and higher doses of LPS did not significantly augment maximum nitric oxide production induced in AMs by LPS alone. Thus, our data suggest that resident rat PMs and AMs resemble elicited mouse PMs in their ability to respond to a single activating stimulus. However, PMs and in some respects AMs differ from the latter by their reduced responsiveness to combinations of activators. Taken together, our results suggest that two exogenous stimuli are not required for full activation of resident macrophages from Sprague-Dawley rats. PMID- 8409755 TI - Antirheumatic gold compounds and penicillamine enhance protein kinase C-mediated activation of the arachidonate-mobilizing phospholipase A2 in mouse macrophages. AB - The effects of antirheumatic gold compounds and D-penicillamine on protein kinase C- and Ca(2+)-mediated activation of arachidonate mobilization and the formation of eicosanoids in mouse macrophages have been investigated. Auranofin (0.2-2 microM) enhanced the response to phorbol ester two- to three-fold, and similar enhancement was caused by aurothiomalate, aurothioglucose, and penicillamine, but only after pretreatment for 1-4 h. The enhanced mobilization of arachidonate was accompanied by increased formation and release of prostaglandin E2 and 6-keto prostaglandin F1 alpha, but not of lipoxygenase metabolites. No such enhancement occurred when the arachidonate-mobilizing phospholipase A2 was activated directly (calcium ionophore A23187). Instead, auranofin caused selective inhibition of calcium ionophore-induced formation of leukotriene C4. Treatment of macrophages with 4 beta-phorbol 12-myristate 13-acetate causes a rapid increase in the phosphorylation and a 1.4-1.8-fold increase in the activity of the 85-kd arachidonate-mobilizing phospholipase A2 as determined in an in vitro assay. The increase in activity was further enhanced by both the gold compounds and penicillamine. These findings indicate that the target for the enhancing effect of the antirheumatic drugs is located between protein kinase C and phospholipase A2 in the signal chain leading to activation of the latter enzyme. PMID- 8409756 TI - Cytochalasin D inhibits lipopolysaccharide-induced tumor necrosis factor production in macrophages. AB - We reported previously that a reorganization of microfilaments can be observed when macrophages are stimulated with lipopolysaccharide (LPS). This reorganization is not detected with current methods in macrophages of an LPS nonresponsive C3H/HeJ mouse. These results suggest that the observed microfilament response might be involved in a macrophage-activating process induced by LPS. To investigate this, we studied the effect of cytochalasin D (CD), which inhibits reorganization of microfilaments, on LPS-induced tumor necrosis factor (TNF) production. A concentration of CD incapable of affecting filamentous actin by itself was used in these experiments. When this concentration of CD was added after LPS stimulation, microfilament reorganization and TNF production were inhibited. The suppressive effect of CD on TNF production was confirmed by the observation that TNF-alpha mRNA expression was also inhibited by CD. This inhibitory effect of CD was not specific to TNF, because the production of interleukin-1 and prostaglandin E2 were also inhibited. These effects of CD were observed only when CD was added within the first 20 min after LPS stimulation. These results suggest that the CD-sensitive microfilament response is essential in the signaling pathway for the production of certain monokines in LPS-stimulated macrophages. PMID- 8409757 TI - Ex vivo clonotype primer-directed gene amplification to identify malignant T cell repertoires. AB - A novel strategy that utilizes input genomic DNA and overcomes limitations encountered with traditional RNA reverse transcription-polymerase chain reaction (PCR) amplification methodology is described to screen for T cell receptor (TCR) repertoires. The methodology has been developed to identify individual T cell clonotypes with regard to their unique receptor beta chain variable/diversity/joining (VDJ) region gene rearrangement. The technique avoids preselection for a given antigen specificity and is therefore independent of artificial bias introduced by in vitro cell population expansion. This technique was used to detect and identify genetically of malignant clones from heterogeneous mononuclear cell populations from an array of hemato-oncological disorders, including mycosis fungoides/Sezary Syndrome, adult T cell leukemia, and large granular lymphoproliferative disease. An initial primary PCR, directed by a TCR-J beta generic primer and a complement of family-specific TCR-V beta primers, defines predominant T cell receptor variable gene usage. Use of a TCR-J beta generic primer supplants the use of a constant region primer anchor and thus eliminates the need to target mRNA. The process of variable gene screening also expedites gene sequencing. By sequencing through the VDJ juxtaposed region, i.e., the third complementarity determinant region, clonotype-specific primers are developed and used in a secondary clonotype primer-directed PCR (CPD-PCR) to detect, with extreme sensitivity and specificity, unique T cell clonal repertoires. Analysis of the products of the CPD-PCR permits the detection of a single malignant cell among one million polyclonal cells and supercedes the constraints of prior studies that provide a limited evaluation of family variable gene repertoire usage. This strategy may be applied in the detection of minimal residual disease, in surveillance after induction of disease-free states, and in analyzing the effectiveness of purging autologous bone marrow of malignant clones. PMID- 8409758 TI - Modulation of the cytoskeleton and intracellular calcium in leukocytes exhibiting a cancer-associated chemotaxis defect. AB - Monocyte chemotaxis is severely depressed in patients with advanced tumors, but the cellular basis for this chemotactic defect is not known. Because the actomyosin cytoskeleton is thought to play a primary role in chemotaxis, we have employed flow cytometry to examine several aspects of the contractile machinery including myosin II, myosin light chain kinase (MLCK), actin, and cytoplasmic calcium in unstimulated and in formylpeptide-stimulated neutrophils and monocytes. Serum-pretreated polymorphonuclear leukocytes (PMNs) and monocytes from healthy blood donors or PMNs and monocytes isolated from tumor patients were studied. Leukocytes pretreated with serum from cancer patients exhibited decreased baseline myosin staining and a vastly different response to formylpeptide stimulation compared with leukocytes pretreated with normal human serum. In contrast, similar amounts of MLCK were observed in neutrophils and monocytes preincubated with normal or cancer serum with or without stimulation with formylpeptide. The fluorescent calcium indicator fluo-3 showed that resting and fMLP-stimulated levels of intracellular calcium were not significantly different in control and cancer serum-pretreated human leukocytes or in leukocytes isolated from tumor patients. Similarly, resting and fMLP-stimulated levels of F-actin in cancer patients' leukocytes as assessed by NBD-phallacidin staining did not differ significantly from those of normal leukocytes. Because the actomyosin cytoskeleton is intricately involved in leukocyte chemotaxis, alterations in the cytoskeleton may dramatically affect cell motility. The cytoskeletal alterations and changes in the response of leukocytes pretreated with cancer patients' serum to formylpeptide stimulation as described here may result in decreased chemotaxis by these cells. PMID- 8409759 TI - Macrophage-tumor cell associations: a factor in metastasis of breast cancer? AB - In human breast carcinomas tumor cells and macrophages are often proximal. We previously reported on the relationship between tumor cell growth and macrophage concentration and report here on the possible involvement of macrophages in the metastatic process. We hypothesize that during the initial stages of metastasis, tumor cells are likely to encounter macrophages and form aggregates. Using a cell culture method that encourages cellular interactions, we found aggregates involving macrophages. Macrophages partly or completely surround other cell types without any apparent ill effect. Units involving macrophages and tumor cells would possess many properties necessary for invasion, which is a normal process for macrophages. Properties such as motility and production of specific enzymes necessary to traverse the extracellular matrix, basement membrane, and endothelial cell barriers may provide an advantage for tumor cells. Physical support and protection from immune recognition during transport of the tumor cell through the vascular system may also be enhanced, and paracrine growth stimulation and angiogenic activity may be provided at the new metastatic site. Verification of these observations in vivo could lead to new directions for limiting breast cancer metastasis. PMID- 8409760 TI - Monocyte adherence to human vascular endothelium. AB - An acute inflammatory response requires that circulating monocytes bind to and migrate across the endothelium of the vessel wall to gain access to inflamed sites. Various mediators of inflammation--cytokines and chemoattractants--have been shown to initiate and regulate the margination and extravasation of monocytes. This review summarizes evidence that the mechanism underlying the initial adhesion of monocytes to normal and cytokine-stimulated endothelial cells and their subsequent transendothelial migration are successive events in monocyte endothelial cell interaction. Special emphasis is given to the current knowledge of the contribution of adhesion molecules belonging to the family of beta 1- and beta 2-integrins, the immunoglobulin supergene family, and the selectins to such cellular interaction. The sequence of events that allow monocytes to attach to, migrate over and finally pass the endothelium is discussed in detail. PMID- 8409761 TI - Plasma cholesteryl ester transfer protein. PMID- 8409762 TI - Effect of dietary n-3 versus n-6 polyunsaturated fatty acids on hepatic excretion of cholesterol in the hamster. AB - Dietary polyunsaturated fatty acids of the n-6 and the n-3 class show differing effects on serum lipids and hepatic lipoprotein metabolism, which could be induced by alterations in hepatocellular cholesterol balance. As both fatty acid classes exert parallel effects on lipoprotein uptake and synthesis of cholesterol in the liver, we studied whether they have differing effects on the excretory pathways for cholesterol. Male Syrian hamsters were fed for 3 weeks low cholesterol diets supplemented (9% w/w) with either saturated (coconut fat), n-6 unsaturated (safflower oil) or n-3 unsaturated fatty acids (fish oil), which shifted the serum lipid levels. N-6 unsaturated fatty acids increased both the synthesis of cholic acid (+57%; P = 0.05) and, in fistula bile, the secretion of cholesterol (+37%; P < 0.05 vs. saturated fatty acids). By contrast, n-3 unsaturated fatty acids did not enhance synthesis of cholic acid or biliary secretion of cholesterol (-30%, NS). The fatty acid pattern of biliary phospholipids was modified according to the major unsaturated fatty acids in the diet. The alterations both in phospholipid fatty acid composition and in secretory ratio of cholesterol to phospholipids and bile acids persisted during controlled secretion of taurocholic acid at increasing rates. In conclusion, hepatic excretion of cholesterol is increased on dietary n-6 unsaturated fatty acids, and low on n-3 unsaturated fatty acids. These two dietary fatty acid classes change differently the fatty acid composition of biliary phospholipids and the secretory ratio of cholesterol to phospholipids and bile acids in bile. PMID- 8409763 TI - Characterization of cholesteryl ester transfer protein inhibitor from plasma of baboons (Papio sp.). AB - Selective breeding has produced families of baboons that accumulate large high density lipoproteins (HDL1) when challenged with a high cholesterol and high fat (HCHF) diet. In the plasma isolated from these high HDL1 baboons there is a factor that decreases the transfer of cholesteryl ester from HDL to lower density lipoproteins. The purpose of these studies was to identify and characterize this inhibitor of cholesteryl ester transfer. A protein with molecular mass of approximately 4 kDa was detected in greater amounts in the plasma lipoproteins of high HDL1 baboons fed the HCHF diet than in plasma lipoproteins of low HDL1 baboons. This 4 kDa protein appeared to associate with apolipoprotein (apo) A-I, resulting in modified apoA-I with an apparent molecular mass of 31 kDa. A small amount of modified apoE was also identified with a molecular mass of 41 kDa. N terminal amino acid sequencing of the 4 kDa peptide identified it as an N terminal fragment of apoC-I. Like apoC-I, the fragment is also a slightly basic protein (pI 7.1). The apoC-I fragment and modified apoA-I presented at equimolar concentrations exhibited similar inhibition of cholesteryl ester transfer protein (CETP) activity in HDL of low HDL1 baboons. On the basis of baboon apoC-I amino acid sequence and the molecular mass of the inhibitor peptide, a peptide corresponding to the N-terminal 38 amino acids of apoC-I was synthesized chemically. This synthetic peptide also inhibited CETP activity in vitro. Rabbit polyclonal antisera prepared against the 38 amino acid synthetic peptide recognized the 4 kDa molecular mass inhibitor protein, apoC-I (6.6 kDa), and the modified apoA-I protein (31 kDa molecular mass) in the plasma lipoproteins of high HDL1 baboons. On the other hand, the antibody detected only apoC-I in the plasma lipoproteins of low HDL1 baboons. The IgG fraction isolated from antiserum raised against the synthetic inhibitor peptide increased cholesteryl ester transfer from HDL of high HDL1 baboons, whereas the IgG antibody against CETP decreased cholesteryl ester transfer from HDL of both high and low HDL1 baboons. These studies suggest that the CETP inhibitor is an N-terminal fragment of apoC I, and this fragment also modifies apoA-I and apoE in the plasma. PMID- 8409764 TI - Dietary supplementation of very long-chain n-3 fatty acids decreases whole body lipid utilization in the rat. AB - Rats were fed lard or n-3 fatty acid-supplemented diets ad libitum to study whole body oxidation of lipid and carbohydrate. One group of male rats was fed 21% fat (by weight) containing 19.5% lard and sufficient amounts of essential fatty acids (1.5%). Another group of rats had 6.5% of the lard replaced by ethyl esters of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The rats were fed these diets for 6-8 weeks. Body weight gain was similar for the two groups and absorption of fat was complete in animals fed both types of fatty acids. Indirect calorimetric measurements, after 3-5 weeks on these diets, by continuous registration of O2 consumption and CO2 formation showed no difference in mean energy expenditure during the experimental period. However, the mean respiratory quotient (RQ) was significantly increased for animals fed the n-3 fatty acid supplemented diet. This was noted both under fasting conditions and after receiving a test meal of similar fatty acid composition for both feeding groups. Thus, mean substrate utilization demonstrated reduced oxidation of fat and increased oxidation of carbohydrate, during fasting as well as fed periods for the n-3 fatty acid group as compared to the lard group. After an additional 2-3 weeks, blood plasma, liver, and muscle samples were collected, and adipocytes and hepatocytes were isolated. Reduced postprandial plasma concentrations of triacylglycerol, phospholipids, unesterified fatty acids, and glycerol were promoted by the n-3 fatty acid diet as compared to lard. Plasma concentration of glucose was slightly increased, and liver and muscle content of glycogen were decreased in the n-3 fatty acid-fed rats. Experiments with isolated adipocytes showed decreased basal lipolysis after feeding n-3 fatty acids for 6-8 weeks for suspended epididymal adipocytes, whereas stimulated lipolysis by isoproterenol (0.1 microM) was higher in both epididymal and mesenteric adipocytes from rats fed n-3 fatty acids as compared to animals fed lard. In addition, epididymal adipocytes from rats fed n-3 fatty acids were significantly smaller than cells from animals fed lard. Hepatic peroxisomal fatty acid oxidation was significantly higher for n-3 fatty acid-supplemented animals, but total fatty acid oxidation was similar in both dietary groups. The hepatic content of triacylglycerol and phospholipids was similar for both diets. These results demonstrate that n-3 fatty acid replacement of a high-fat diet containing mostly saturates and monoenes for several weeks promotes reduced use of fat as energy source. This may be explained by decreased plasma concentration of unesterified fatty acids.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8409765 TI - Human very low density lipoprotein structure: interaction of the C apolipoproteins with apolipoprotein B-100. AB - Very low density lipoproteins (VLDL) are a heterogenous population of particles differing in size and composition. Heparin-Sepharose chromatography yields three VLDL subfractions. Two subfractions, VLDLNR-1 and VLDLNR-2, which are not retained by heparin, contain little or no detectable apolipoprotein (apo)E. According to negative stain electron microscopy, VLDLNR-1 is slightly larger than VLDLNR-2. The third fraction, VLDLR, is composed of smaller particles that are retained by the heparin-Sepharose and contain apoE. The C apolipoproteins of the respective VLDL subfractions transfer to 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC) single bilayer vesicles giving three subfractions designated VLDLNR-1-C, VLDLNR-2-C, and VLDLR-C. The protein, phospholipid, and cholesterol (free + esterified) contents decrease in the order VLDLR > VLDLNR-2 > VLDLNR-1. Triglyceride content decreases in the opposite order. POPC treatment of each VLDL subfraction increases the phospholipid and decreases the protein, triglyceride, and cholesteryl ester contents, while free cholesterol remains unchanged. According to immunological analysis of each subfraction with well-characterized monoclonal antibodies, the accessibility of some epitopes of apoB-100 on VLDL is changed by POPC treatment. Electron-microscopic analysis of POPC-treated VLDL subfraction reveals vacancies on the surfaces of each particle. VLDLNR-1, VLDLNR 2, and VLDLR are resistant to thrombin cleavage, whereas the lipoproteins lacking C apolipoproteins are not. Thrombin cleavage (8 h) of apoB-100 of VLDLNR-2-C and VLDLR-C gives two fragments, T1 and T2, that are converted to smaller fragments only after prolonged treatment. In contrast, apoB-100 of VLDLNR-1-C is converted into small fragments after 8 h thrombin treatment. These results suggest that removal of apoCs affects the accessibility and conformation of apoB-100 in the individual VLDL subfractions in the region near residue 3249, which is the primary thrombin cleavage site and the epitope of monoclonal antibody 4C11. PMID- 8409766 TI - Nucleotide sequence encoding the carboxyl-terminal half of apolipoprotein B from spontaneously hypercholesterolemic pigs. AB - Previous studies from this laboratory characterized the hypercholesterolemia of pigs with a mutant allele of apolipoprotein B (apoB), designated Lpb5. This apoB allele is associated with low density lipoprotein (LDL) particles deficient in binding to the LDL receptor. To identify potential causative mutations in Lpb5 DNA, 10.6 kb of genomic DNA, encoding the carboxyl-terminal 58% of apoB were sequenced from the Lpb5 allele and from an allele encoding phenotypically normal apoB. Comparison of the two DNA sequences revealed 33 polymorphisms, 13 of which resulted in amino acid polymorphisms. To determine whether any of the amino acids at the polymorphic positions in Lpb5-encoded apoB were unique to that isoform, those positions were sequenced in four other pig apoB alleles encoding phenotypically normal apoB. None of the amino acids were by themselves uniquely encoded by the Lpb5 allele. However, a unique haplotype consisting of Asp3164 in conjunction with Ala3447 distinguished the Lpb5-encoded apoB from all other allelic isoforms sequenced in this region. To gain insight into changes in the tertiary structure of the mutant apoB, 13C-NMR analysis of LDL reductively methylated with [13C]-formaldehyde was performed. LDL has lysine residues that titrate at pH 10.5 and others that titrate at pH 8.9. The latter residues are thought to include those involved in the interaction of LDL with the LDL receptor. LDL from Lpb5 pigs possessed a smaller proportion of lysine residues titrating at pH 8.9 than did LDL from non-Lpb5 pigs, suggesting that the Lpb5 encoded apoB is altered in a manner affecting the microenvironment of particular lysine residues. PMID- 8409767 TI - Alternative splicing of mutant LDL-receptor mRNA in an Italian patient with familial hypercholesterolemia due to a partial deletion of LDL-receptor gene (FHPotenza). AB - An analysis of LDL-receptor gene was performed on an Italian patient with heterozygous familial hypercholesterolemia. Restriction enzyme analysis showed that the proband was heterozygous for a deletion of 4.5 kb spanning the 5' end of exon 13 (45 nucleotide residues) to intron 15. Amplification of genomic DNA, using polymerase chain reaction (PCR), followed by direct sequencing, showed that this deletion was identical to the one reported by Lehrman et al. (1986. Proc. Natl. Acad. Sci. 83: 3679-3683). As only the normal LDL-receptor mRNA was detectable in proband fibroblasts by Northern blot, we used reverse transcription PCR to amplify the mutant mRNA using primers complementary to exon 6 (sense) and exon 18 (antisense). The amplification of control cDNA resulted in a single fragment of 1725 nucleotides containing the normal sequence. The amplification of cDNA from the proband produced the 1725-nucleotide fragment (as in the control) and three additional fragments (F1, F2, and F3) of smaller size. The direct sequence showed that in the F1 fragment exon 12 was joined to exon 16; in the F2 fragment exon 12 was joined to exon 17; and in the F3 fragment exon 11 was joined to exon 16. Thus, the deletion-bearing allele generated three mRNAs, two of which resulted from alternative splicings leading to the skipping of exons 16 and 12, respectively. It is expected that the translation of these mutant mRNAs will generate three aberrant proteins, the synthesis of which should be negligible in view of the very low content of the corresponding mRNAs. PMID- 8409768 TI - Apolipoprotein B mRNA editing in 12 different mammalian species: hepatic expression is reflected in low concentrations of apoB-containing plasma lipoproteins. AB - Two different isoproteins are encoded by the apolipoprotein (apo) B gene, apoB-48 and apoB-100. ApoB-48, core component of intestinally derived chylomicrons, has an accelerated plasma turnover as compared with the full-length protein apoB-100. A posttranscriptional modification of the apoB mRNA by conversion of cytidine into uridine at nucleotide position 6666 changes the genomically encoded glutamine codon CAA at amino acid residue 2153 into a translational stop codon UAA. This mRNA editing explains the formation of the truncated isoform apoB-48. In the present investigation editing of apoB mRNA in liver and intestine from 12 different mammalian species was measured by a quantitative primer extension analysis of reverse-transcribed and polymerase chain reaction- (PCR) amplified apoB mRNA in order to determine whether i) editing of apoB mRNA is generally restricted to the intestine or may also be found in the liver of other species than rodents, and ii) hepatic expression of apoB mRNA editing influences lipoprotein concentrations in plasma. Intestinal apoB mRNA was edited at high levels in all species, 40% in sheep, 73% in horse, 82% in pig, 84% in dog, 84% in cat, 87% in guinea pig, 88% in rat, 89% in mouse, and > 90% in human, monkey, cow, and rabbit. In liver apoB mRNA was edited to 18% in dog, to 43% in horse, to 62% in rat, and to 70% in mouse. Low levels of editing below 1% were detected in liver of rabbit and guinea pig. In contrast, hepatic apoB mRNA from human, monkey, pig, cow, sheep, and cat liver was not edited. The results of the primer extension analysis were confirmed by cloning and sequencing of the PCR products from dog, horse, cat, guinea pig, sheep, and cow for all of which the apoB cDNA sequence had not been established by previous investigations. Primer extension analysis of apoB mRNA from dog intestine and dog liver indicated C/U editing at C6655 in addition to C6666. Cloning and sequencing of apoB cDNA from dog liver and intestine confirmed additional C/U editing at C6655 which changes ACA for threonine at amino acid residue 2149 into AUA for isoleucine. Synthesis and secretion of apoB-48-containing lipoproteins from liver was demonstrated by pulse labeling of freshly isolated horse hepatocytes and immunoprecipitation with apoB specific antibodies or density gradient ultracentrifugation. The concentrations of VLDL, LDL, and HDL in all species were determined after fractionation by density gradient ultracentrifugation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8409769 TI - Degradation of plasma membrane phosphatidylcholine appears not to affect the cellular cholesterol distribution. AB - To clarify the role of possible cholesterol/phosphatidylcholine interactions in cellular cholesterol distribution, we have used a phosphatidylcholine-specific phospholipase C from Bacillus cereus to degrade the cell surface phosphatidylcholine of cultured human fibroblasts. Of cellular phosphatidylcholine, approximately 15% was susceptible to degradation by the phospholipase. In spite of the dramatic redistribution of cellular cholesterol that can be observed after sphingomyelin depletion, the degradation of cell surface phosphatidylcholine did not affect the distribution of cholesterol in fibroblasts. In cholesterol-depleted cells as well as in cholesterol-loaded cells, the size of the cell surface cholesterol pool (susceptible to cholesterol oxidase) remained unchanged after phosphatidylcholine degradation. The rate of cholesterol esterification with [3H]oleic acid and the rate of [3H]cholesterol efflux from fibroblasts to high density lipoproteins also remained unchanged after degradation of plasma membrane phosphatidylcholine. An increase in the level of [3H]cholesterol efflux to high density lipoproteins was observed after degradation of plasma membrane sphingomyelin with exogenous sphingomyelinase, in contrast to earlier reports, where no such effect was observed. The results suggest that interactions between cholesterol and phosphatidylcholine in the fibroblast plasma membranes are less important than cholesterol/sphingomyelin interactions for the asymmetric distribution of cellular cholesterol. PMID- 8409770 TI - Functional characterization of a chimeric lipase genetically engineered from human lipoprotein lipase and human hepatic lipase. AB - Lipoprotein lipase (LPL) and hepatic lipase (HL) mediate the hydrolysis of triglycerides and phospholipids present in circulating lipoprotein particles and are essential for normal lipid metabolism. Both enzymes have a similar primary amino acid structure and share requirements for intact catalytic, lipid binding, and heparin binding domains. However, LPL and HL exhibit different substrate specificities and cofactor requirements. In order to characterize the functional domains necessary for LPL activity, a chimeric lipase consisting of the amino terminal 314 amino acids of human LPL and the carboxyl-terminal 146 amino acids of human HL was synthesized by joining the cDNA of both lipases at the 5'-end of exon 7. Northern blot hybridization and Western blot analyses revealed the size of the chimera mRNA and protein to be approximately 1.5 kb and 55 kDa, respectively. The chimeric enzyme hydrolyzed both long chain and short chain fatty acid triacylglycerols and had catalytic properties that were similar to lipoprotein lipase. Thus, apolipoprotein (apo)C-II was required for maximal lipase activity, and high salt concentration abolished the ability of the chimera to hydrolyze triolein even in the presence of apoC-II. A monospecific anti-HL polyclonal antibody interacting with the C-terminal HL-derived domain of the chimeric enzyme abolished the enzyme's ability to hydrolyze triglyceride emulsion but not tributyrin substrates. Analysis of the heparin binding properties of the chimeric enzyme using heparin-Sepharose affinity chromatography revealed an elution pattern which was intermediate between that of lipoprotein and hepatic lipase. In summary, we have characterized the functional properties of an LPL-HL chimeric enzyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409771 TI - Effect of dietary cholesterol on cholesterol synthesis in breast-fed and formula fed infants. AB - The fractional synthesis rate (FSR) of cholesterol was measured in 6 breast-fed and 12 formula-fed infants (ages 4 to 5 months) using the 2H2O method. The breast fed infants had higher cholesterol intakes (18.2 +/- 4.0 vs. 3.4 +/- 1.8 mg/kg per day, P = 0.001), plasma total cholesterol (183 +/- 47 vs. 112 +/- 22 mg/dl, P = 0.013), and plasma low density lipoprotein (LDL)-cholesterol (83 +/- 26 vs. 48 +/- 16 mg/day, P = 0.023) than the formula-fed infants (6.9 +/- 2.6 vs. 2.1 +/- 0.6%/day, P < 0.001). Among all infants, there was a significant inverse relationship (P = 0.002, r = 0.66) between the FSR of cholesterol and dietary cholesterol intake. Our findings indicate that the greater cholesterol intake of the breast-fed infants was associated with elevated plasma LDL-cholesterol concentrations and that cholesterol synthesis in human infants may be efficiently regulated via HMG-CoA reductase when infants are challenged with high intakes of dietary cholesterol. PMID- 8409772 TI - Effects of dietary fats from animal and plant sources on diet-induced fatty streak lesions in C57BL/6J mice. AB - This study was designed to determine the effects of a variety of naturally occurring saturated fats on aortic lesion formation in C57BL/6J mice that are susceptible to diet-induced fatty streak lesions. Groups of female mice were randomly assigned to one of seven treatment groups and were fed diets containing 15% (w/w) hydrogenated coconut oil, hydrogenated soy oil, hydrogenated palm oil, cocoa butter, lard, tallow, or dairy butter, 1% cholesterol, and 0.5% cholic acid. Plasma lipid levels were measured to determine whether lesion formation was related to specific changes in these parameters. Lesions, which were observed in all groups of mice, ranged from 420 to 3220 microns2/aortic cross section. Lesion area was positively correlated to the percentage of saturated fatty acids contained in the fat sources and the ratio of combined VLDL plus LDL-cholesterol to HDL-cholesterol and inversely correlated to monounsaturated fatty acids content and to HDL-cholesterol levels. Results from this study demonstrate that inbred mice may provide a good model for dissecting the genetic basis for the differential atherogenic responses to diet-induction and for studying the effects of dietary factors on aortic lesion development. PMID- 8409773 TI - Modification of spleen phospholipid fatty acid composition by dietary fish oil and by n-3 fatty acid ethyl esters. AB - We have compared the effects of diets containing purified ethyl esters of either eicosapentaenoic acid, docosahexaenoic acid, or a mixture of both of these compounds, with diets containing either purified fish oil or beef tallow on spleen phospholipid fatty acid composition. Autoimmune mice, the (NZB x NZW)F1 strain, were fed with experimental diets for 14 weeks, after which spleen phospholipids were extracted and separated into classes by HPLC, and the alkenylacyl, alkylacyl, and diacyl subclasses of glycerylphosphatidylethanolamine and glycerylphosphatidylcholine were resolved as their benzoyl esters by HPLC. Fatty acids were analyzed by capillary gas-liquid chromatography of their methyl esters. Each of the marine lipid diets suppressed n-6 fatty acids and elevated n 3 fatty acids in all phospholipids. The eicosapentaenoic acid ethyl ester diets led to high levels of eicosapentaenoic acid and docosapentaenoic acid (C22:5n-3), but little or no increase in docosahexaenoic acid. The docosahexaenoic acid ethyl ester diets elevated docosahexaenoic acid, docosapentaenoic acid, and eicosapentaenoic acid in all phospholipids, indicating that extensive retroconversion of 22 carbon n-3 fatty acids had occurred. These results document changes in the fatty acid composition of mammalian phospholipids that are induced by dietary fish oil triglycerides and by dietary long chain n-3 fatty acid ethyl esters. PMID- 8409774 TI - Suppression of autoimmune disease by dietary n-3 fatty acids. AB - Previous studies have demonstrated that dietary fish oil preparations have anti inflammatory effects in humans and in experimental animals, but the individual components of fish oils that are responsible for their anti-inflammatory effects have not been documented. We therefore investigated in (NZB x NZW)F1 mice, a model for human systemic lupus erythematosus, the effects of diets containing ethyl esters of two purified n-3 fatty acids, eicosapentaenoic acid (EPA-E) and docosahexaenoic acid (DHA-E), a refined fish oil triglyceride (FO) which contained 55% n-3 fatty acids, and beef tallow (BT) which contains no n-3 fatty acids. The diets were initiated prior to the development of overt renal disease at age 22 weeks, and continued for 14 weeks. The extent of the renal disease was quantified by light microscopy and by proteinuria. Diets containing either 10 wt% FO, 10% EPA-E, or 6% or 10% DHA-E alleviated the severity of the renal disease, compared to the BT diet, whereas diets containing either 3% or 6% EPA-E or 3% DHA E were less effective. Two diets containing approximately 3:1 mixtures of EPA-E and DHA-E alleviated the renal disease to a greater extent than expected for either of these fatty acids given singly. We believe that these experiments provide the first demonstration of anti-inflammatory effects of individual dietary n-3 fatty acids. The results also indicate that the anti-inflammatory effects of fish oils depend on synergistic effects of at least two n-3 fatty acids. PMID- 8409775 TI - Changes in biliary lipid secretion during normal development and diurnal cycling in the rat. AB - Biliary lipid secretion in conjunction with hepatic cholesterol synthesis was determined in normal male rats at 4, 5, 7, and 9 weeks of age, during a period of linear growth and a near fourfold increase in liver size. Studies were performed both at periods of low (mid-light cycle) and high (mid-dark cycle) hepatic cholesterol synthesis. Biliary bile salt, phospholipid, and cholesterol secretion (per g of liver) markedly decreased with an increase in liver size. Whereas the secretion of bile salts and phospholipid was not significantly different in mid dark and mid-light periods for animals of the same age, cholesterol secretion was greater in the mid-dark than in the mid-light period at 5, 7, and 9 weeks of age. Relationships between biliary cholesterol secretion and bile salt and phospholipid secretion differed at mid-dark and mid-light periods, as follows: cholesterol secretion was not significantly related to bile salt secretion at mid dark (r = 0.49, P > 0.05) but was related at mid-light (r = 0.73, P = 0.003); and although cholesterol secretion was significantly related to phospholipid secretion at mid-dark, this relationship was not nearly as strong as at mid-light (P < 0.005, comparing r = 0.95 at mid-light with r = 0.53 at mid-dark). In contrast, at mid-dark, biliary cholesterol secretion was strongly related to hepatic cholesterol synthesis (r = 0.84, P < 0.0001) whereas at mid-light the two were not significantly related (r = 0.13, P > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409776 TI - Effect of recombinant-tissue plasminogen activator, low molecular weight urokinase and unfractionated heparin on platelet aggregation. AB - We studied the interaction of two thrombolytic agents, recombinant-tissue plasminogen activator (r-TPA) and low molecular weight urokinase (UK), with platelet aggregation in the absence or presence of unfractionated heparin (UH). With platelet rich plasma (PRP), a dose dependent inhibition was seen for both r TPA and UK. However, this effect was more evident in the presence of r-TPA. UH did not modify this effect. However, it enhanced platelet aggregation induced by ADP and decreased the aggregation induced by collagen (COL) as already shown by us. With washed platelets, only r-TPA decreased platelet aggregation in a dose dependent manner in the presence of COL and only at the highest dose (100 micrograms) in the presence of TH. The presence of ten units of plasminogen (PLG) together with 10 micrograms of r-TPA or 1250 units of UK totally inhibited TH induced platelet aggregation. UH reversed this effect. In contrast, when COL was the aggregating agent, the inhibition of platelet aggregation in presence of PLG, seems to be further increased by UH. Since UH is an adjunct of thrombolytic therapy to prevent rethrombosis, this double edged sword could partially explain the lack of therapeutic effect in some patients. PMID- 8409777 TI - Serum levels of pentoxifylline and its metabolites in non-tumor-bearing mice after chronic oral pentoxifylline administration. AB - Pentoxifylline lowers blood viscosity and increases erythrocyte flexibility in patients with atherosclerosis, thus increasing tissue oxygen delivery. Since tumor neo-vessels are associated with tissue hypoxia, which contributes to failure of radiation therapy, pentoxifylline might also be useful as a radiation sensitizer. Such drugs are often evaluated in the murine model, but serum levels of pentoxifylline and its metabolites have not been determined in the mouse after chronic oral dosing. We investigated this by administering pentoxifylline via liquid diet or solid diet to young adult male CF1 mice for one to six weeks and assaying plasma for the parent drug and its metabolites. At one, three, and six weeks, plasma levels of pentoxifylline and major derivatives were consistently detectable. Mice remained healthy during this period, indicating that ingestion of large amounts of this drug is well tolerated. PMID- 8409778 TI - Persistence of enterovirus RNA in muscle biopsy samples suggests that some cases of chronic fatigue syndrome result from a previous, inflammatory viral myopathy. AB - Molecular hybridization using an enterovirus group specific probe detected virus RNA in muscle biopsy samples from 25 of 96 cases of inflammatory muscle disease and similarly from 41 of 158 cases of postviral fatigue syndrome (PFS). Enterovirus RNA was detected in only two of 152 samples of control muscle. The inflammatory myopathy group comprised patients with polymyositis (PM), juvenile dermatomyositis (JDM) or adult dermatomyositis (DM), and all showed the presence of an inflammatory infiltrate and fiber necrosis on histological examination of a muscle biopsy sample. In contrast, muscle samples from the PFS group were histologically normal except for non-specific changes such as occasional single fiber atrophy. By analogy with enteroviral myocarditis, which can progress to a post-inflammatory disease with persistence of virus in myocardium and disposes to the rapid development of dilated cardiomyopathy, we propose that PFS syndrome may be a sequela of a previous inflammatory viral myopathy. PMID- 8409779 TI - Child abuse, osteogenesis imperfecta and the grey zone between them. AB - We report a child who was thought to suffer a non-accidental injury. The parents were unable to convince the child abuse team of their innocence. The eruption of lucent teeth established the diagnosis of osteogenesis imperfecta type IVB. PMID- 8409780 TI - Relationship between zinc and obesity. AB - Zinc has important effects on metabolism, and on the thermoregulation of obese individuals. The aim of our investigation was to evaluate the serum zinc levels in obese patients before and after severe hypocaloric diets and to evaluate its correlation with the body mass index (BMI). Patients followed a severe hypocaloric diet (737 Kcal) for 60 days. Serum Zn levels and BMI were evaluated. Zn levels in obese patients were significantly (p < 0.01) lower than in controls, whereas the BMI values were significantly greater. At the end a severe hypocaloric diet, serum Zn and BMI levels returned to normal values. Our data show a possible relationship of the serum Zn levels with the anabolic and catabolic mechanisms in obesity, although the exact metabolic role of this bio element remains unclear. PMID- 8409781 TI - Plasma fibrinogen and platelet count in stroke. AB - Plasma fibrinogen levels and platelet counts were evaluated in 30 patients with acute ischemic cerebral infarction (CI) three and nine days after the onset of symptoms. Hyperfibrinogenemia (379.4 +/- 80.3 vs 327.3 +/- 48.3 mg/dL of controls, p < 0.005), a reduction of platelet count (207.133 +/- 48.388 vs 288.375 +/- 61.373 x 10(9)/L of controls, p < 0.001), and an inverse correlation (r = -0.41, p < 0.05) between the two parameters were observed on day three. On day nine, platelet counts normalized while plasma fibrinogen levels slightly increased; the inverse correlation no longer occurred. The results suggest that in the earliest phase of stroke plasma fibrinogen levels may condition the extent of platelet accumulation or consumption in the ischemic area. This confirms in vivo that the platelet aggregation process is strictly dependent on fibrinogen concentration. PMID- 8409782 TI - Tumor resection attenuated the impaired tolerance of glucose in patients with asymptomatic pheochromocytoma. AB - A 51-year-old male was admitted to our hospital because of glucosuria and an abdominal tumor. While blood pressure levels were maintained within normal ranges, serum adrenaline and noradrenaline levels and the urinary excretion of total metanephrine increased. The oral glucose tolerance test (75g-OGTT) showed a diabetic pattern. Venography and blood sampling from the adrenal vein showed the existence of a right adrenal tumor. Right adrenalectomy normalized serum catecholamine levels and the urinary excretion of metanephrine. Simultaneously, fasting glucose levels returned to normal. The present case suggested that increased metanephrine secretion may affect only pancreatic insulin secretion, but not blood pressure levels in the patient with asymptomatic pheochromocytoma. PMID- 8409783 TI - Edema due to altered sweating function. AB - We report a case with idiopathic edema accompanied with excessive sweating on bathing. The subject is a 26 year-old male. He was admitted to our hospital because of pretibial edema. He complained of excessive sweating only after daily bathing. Sweating on daily bathing reduced pretibial edema after long-term standing. Daily urine volume decreased by about 200 to 500 mL/day on bathing everyday. Plasma renin, aldosterone and bradykinin levels increased. Subcutaneous injection of pilocarpine increased sweating around the injection site. The cessation of daily bathing restored plasma renin, aldosterone and bradykinin levels; urine volumes increased. However, pretibial edema appeared and body weight increased by 8.5 Kg. Serum triiodothyronine (T3), thyroxine (T4) levels and basal metabolic rate (BMR) decreased. Circulating plasma volume increased with the cessation of bathing. Alterations in the autonomic nervous system may be involved in the appearance of general edema through increased plasma volume due to altered sweating function. PMID- 8409784 TI - Interexaminer reliability of eight evaluative dimensions of lumbar segmental abnormality: Part II. AB - OBJECTIVE: The objectives of this study were to assess the interexaminer agreement of palpation for soft tissue and osseous pain along with visual observations in the lumbar spine. Second, the interexaminer agreement of dermothermograph and surface electromyographic (EMG) scans of the lumbar spine were assessed. Third, to perform these evaluations on symptomatic low back patients. Finally, the most reliable measurements were combined in a multidimensional index of segmental lumbar abnormality, which was assessed for interexaminer agreement. DESIGN: This is an interexaminer reliability study of commonly used palpatory and instrumentation procedures used to assess lumbar segmental abnormality. SETTING: This study was conducted at Pain Assessment and Rehabilitation Center (PARC) and the Center for Clinical Studies (CCS) at Northwestern College of Chiropractic. PATIENTS: The patients involved in this study were symptomatic at the time of examination. The patients were recruited from the CCS clinic and PARC. RESULTS: Palpation for osseous pain produced kappa coefficients ranging from .48-.90. Palpation for soft tissue pain produced kappa coefficients that ranged from .40-.79 and the kappas for visual observation ranged from .34-.84. The dermothermograph and surface EMG scanner were also assessed with the kappa coefficient for their reliability in assessing lumbar segmental abnormality. The kappa coefficients ranged from -.13 to .59 for the surface EMG and 0- .63 for the dermothermograph measurements. Intraclass correlation coefficients for the surface EMG measurements ranged from .20-.55 and the dermothermograph measurements ranged from .01-.55. Palpation for pain (osseous and soft tissue) and visual observation were included in the multidimensional index of abnormality. The interexaminer agreement of detecting a manipulable lesion was evaluated by designating a lesion present with a positive two out of three tests. Kappa coefficients for the multidimensional index of lumbar abnormality ranged from a low of .05 to a high of .52. CONCLUSIONS: Palpation for pain (osseous and soft tissue) and visual observation produced good to excellent interexaminer agreement and were included in the multidimensional index of abnormality. The interexaminer agreement of surface EMG scans and dermothermograph measurements were poor and considered to be clinically unacceptable, thus were not included in the multidimensional index. Palpation for pain is the only spinal assessment procedure to show consistent reliability in a number of studies. PMID- 8409785 TI - United States Department of Veterans Affairs Chiropractic Services Pilot Program evaluation study SDR #86-09: a critique. AB - The following critique of the Chiropractic Services Pilot Program (SDR #86-09) focuses on two major issues: the terms of reference established for the study and the research constraints that arose from either the terms of reference or their interpretation; the technical design and execution of the research. The review suggests that the constraints invalidated the study and ensured that no comparisons are possible between chiropractic and medical care for VA patients based on these results. The constraints resulted in the use of a nonexperimental design, distinct samples being chosen and nonequivalent care settings being compared. The critique also reveals that in each of the design steps (eligibility criteria, sampling, protocols, data collection, analysis, interpretations) there were serious methodological flaws. These ensured that the two populations being compared (chiropractic patients versus medical patients) were in fact noncomparable. In terms of the economic cost comparisons, the design guaranteed the comparison was unfair, pitting a private, fee-for-service chiropractic practice against a not-for-profit, managed-care, federally regulated and budgeted institution. Furthermore, the allocation of costs to the two groups was done inaccurately. The critique concludes that the results are not valid, they cannot be used for generalizing, they cannot be used for statistical analysis and they should not be used to establish policy. The research design and the methodological flaws meant that the objectives of the study could not be met. PMID- 8409786 TI - Chiropractic biophysics lateral cervical film analysis reliability. AB - OBJECTIVE: To determine the degree to which the geometric line drawings used in Chiropractic Biophysics Technique (CBP) on lateral cervical radiographs are reliable. DESIGN: A blind, delayed repeated measures design was used. Three examiners were presented radiographs in random order. All identifying marks were removed prior to each examiner's individual marking and measurement. Each examiner was blinded as to how the previous examiners marked and measured the radiographs. SETTING: Primary care private chiropractic clinic. PATIENTS: PARTICIPANTS: Sixty-five subject films were provided from the patient records of a primary care private chiropractic clinic. The 65 radiographs qualified for inclusion in the study based on two criteria: C1 through C7 had to be clearly visible, and there had to be no identifying artifacts. MAIN OUTCOME MEASURES: Anterior head translation in millimeters, atlas plane to horizontal, Ruth Jackson's cervical stress lines, and five relative rotation angles for C2-C3, C3 C4, C4-C5, C5-C6, C6-C7. Inter- and intrareliability of the three examiners were statistically analyzed. RESULTS: Intraexaminer for a) C1 to horizontal reliability was .98-.99 with confidence intervals of .96-.99, b) absolute rotation angle from C2 to C7 reliability was .82-.95 with confidence intervals of .80-.99, c) anterior head translation [+Sz] reliability was .86-.99, with confidence intervals of .74-.99, d) relative rotation angle reliability ranges were (C2-C3) .99, and (C3-C4) .98-.99, (C4-C5) .88-.99, (C5-C6) .80-.99, and (C6 C7) .94-.98. Interexaminer reliabilities across examiners ranged from a) Winer:.89-.99 and b) Bartko: .72-.96. CONCLUSIONS: The reliabilities for intra- and interexaminer were all greater than .70, indicating that these measurements in CBP technique would be considered accurate enough to provide measurements for future clinical studies. The data indicated that the C6-C7 relative rotation angle was the least reliable measurement. This might be due to the very small angles found at this level. PMID- 8409787 TI - Chiropractic diagnosis and treatment of closed head trauma. AB - OBJECTIVE: The objective of this article is to review the current literature relating to the chiropractic diagnosis and treatment of closed head trauma. It outlines the clinical exam, offers a diagnostic protocol and describes current chiropractic management and treatment of acute and chronic closed head trauma. Particular importance is placed on the need to differentiate between mild, moderate and severe head injury. Treatment protocols are elucidated for cerebral concussion and a rationale proposed for the management and treatment of posttraumatic concussion syndrome. DATA SOURCES: Information was obtained from English language chiropractic, medical and scientific journals as well as chiropractic and medical textbooks. The CHIROLARS data retrieval system was used (year 0-1992) as was the MEDLINE data base (1988-1992). Head trauma, head injury, headache, concussion, vertigo, posttraumatic syndrome and whiplash injury were the indexing terms used. CONCLUSION: The doctor of chiropractic is often the first practitioner a patient will see following a motor vehicle accident, sports injury or other acute trauma. The chiropractor is also the practitioner a patient seeks for help after suffering for months with chronic posttraumatic concussion syndrome. It is important that we have a protocol for effectively managing both acute and chronic closed head injury. PMID- 8409788 TI - Electrodiagnostic testing in back and extremity pain. AB - OBJECTIVE: The usefulness of electrodiagnostic testing by the primary care provider is shown in two cases of suspected compression-type neural lesions of the lumbar spine. CLINICAL FEATURES: A 54-yr-old female with acute lumbar spine pain that radiated into the hip and a 26-yr-old male with sharp gluteal pain that radiated into the thigh and ankle were admitted into an inpatient care facility for intensive therapy. Plain film radiographs were obtained initially. In addition, electrodiagnostic testing was performed to evaluate the L5 and S1 nerve roots, which suggested compressive-type lesions. Non-enhanced CT of the lumbar spine was performed and revealed central disk herniation or protrusion in each case. INTERVENTION AND OUTCOME: One patient exhibiting central compression signs of bladder dysfunction was referred for medical intervention. The remaining patient received flexion-distraction type of chiropractic manipulation with physiologic therapeutics and was discharged to outpatient care after 16 days. CONCLUSIONS: Electrodiagnostic testing can provide the primary care provider the data needed to make an informed decision regarding advanced imaging studies and to institute appropriate therapy or to intelligently refer a patient for follow up. PMID- 8409789 TI - Nonoperative management of lumbar spinal stenosis. AB - OBJECTIVE: To describe the successful treatment of a patient with lumbar spinal stenosis utilizing nonoperative procedures. CLINICAL FEATURES: A 76-yr-old male with a chief complaint of low back pain and left lower extremity pain demonstrated the following per history and physical examination: 1. A right antalgic shift. 2. Restricted lumbar range of motion with provocation of left lower extremity pain during extension. 3. Generalized lumbar spondylosis as revealed on plain film X rays. 4. MRI confirmed lumbar stenosis. A diagnosis of lumbar spinal stenosis secondary to spondylosis was made. INTERVENTION AND OUTCOME: Twelve treatments of flexion-distraction manipulation, deep tissue massage, ultrasound, therapeutic exercise, heel lift, and modification of activities of daily living. He was discharged from care asymptomatic in 3 wk. Objective improvement was also noted. CONCLUSIONS: Conservative treatment designed to increase lumbar flexion, thus increasing lumbar spinal canal volume, has a positive influence on the diminution of neural ischemia and its resultant neural dysfunction. Additional research is needed to elucidate these concepts. PMID- 8409790 TI - Resolution of infantile Erb's palsy utilizing chiropractic treatment. AB - OBJECTIVE: To present a case study of infantile Erb's palsy that responded to conservative chiropractic care. CLINICAL FEATURES: A 5-wk-old infant boy suffered from a limp left arm. A clinical diagnosis of Erb-Duchenne palsy was made. Birth records were obtained to further substantiate the diagnosis. INTERVENTION AND OUTCOME: The patient received specific chiropractic adjustments to the mid cervical spine and muscle stimulation therapy with an upper extremity exercise program. The Erb's palsy resolved with only a mild residual "waiters tip" deformity within 2 months. CONCLUSION: In this case, chiropractic adjustments with muscle stimulation therapy is suggested as an effective treatment for Erb's palsy. Further studies are required to better understand chiropractic's effectiveness in Erb's palsy cases. PMID- 8409791 TI - Alternative philosophical and investigatory paradigms for chiropractic. PMID- 8409792 TI - Quackery vs. accountability in the marketing of chiropractic. PMID- 8409793 TI - Chiropractic magazines. PMID- 8409794 TI - Asymmetry of occipital condyles: a computer-assisted analysis. PMID- 8409795 TI - Cervicogenic dysfunction in muscle contraction headache and migraine: a descriptive study. PMID- 8409796 TI - The mammography project. PMID- 8409797 TI - Highlights of the May MAG/Aetna Medicare Advisory Committee. PMID- 8409798 TI - Guidelines for identifying and helping abused patients. PMID- 8409799 TI - Gateway House: a safe place. PMID- 8409800 TI - "Growing healthy" what's happening around Georgia? PMID- 8409801 TI - A pilot project for Georgia "growing healthy" in Ware County. PMID- 8409802 TI - "The hands that heal" a history of the practice of medicine in northwest Georgia. PMID- 8409803 TI - The Atlanta Project. Hope for children. PMID- 8409804 TI - Divorce after 40 years. PMID- 8409805 TI - Facing medicine's challenges: proverbs, music, and politics. PMID- 8409806 TI - The AMA and health system reform. AMA update from the front lines. PMID- 8409807 TI - A method for selecting data subsets from large medical data bases. AB - Because of the exponential increase in health care expenditures, attention has been drawn to the cost and effectiveness of health care delivered. The use of electronic data bases as a means for assessing health care effectiveness has been advocated as powerful computer systems have become commonplace. The need to work efficiently with extremely large data bases presents difficult challenges to researchers who wish to focus on specific aspects of the data. This paper describes a computer program written to facilitate the analysis of large medical data sets by providing a straightforward way to specify and select data subsets from the aggregate data base. PMID- 8409808 TI - Clinical laboratory test reference (CLTR). AB - As the healthcare system undergoes a transformation in scope and funding, there remain many unfinished projects which will be essential for the next generation of automated medical support services. The most demanding and labor intensive tasks for this new frontier deal with the accumulation of knowledge which can be used as a clinical database to support supervisory functions in a physician operated interactive care delivery environment. These databases will contain the worlds accumulated knowledge in specialized areas. They will be organized by topic or clinical service, and have significant impact on the quality of care as well as medical malpractice exposure. This article will describe a clinical pathology database that has been adapted for medical practice. The database contains information about laboratory tests and their interpretation. The data is structured for rapid reading and has references where indicated. The database can be used in a stand alone program or integrated into an information system within an application program. The files are reviewed on a continuing basis and quarterly updates are made available to subscribers. PMID- 8409809 TI - The effects of electronic mail on communication in two health sciences institutions. AB - A study was done during 1991-1992 to determine the perceived impact of electronic mail (E-mail) relative to other forms of communication in health sciences institutions. E-mail subscribers at two major health sciences institutions were sent 2919 surveys, and 823 (28%) completed survey instruments were returned. A significant positive impact of E-mail was found relative to other forms of communication (e.g., paper, phone) with regard to E-mail messaging, response rates, influence, value, formality, perceptions, errors in communication, cost effectiveness, communication style, and other factors. Areas where no differences were found between communication mechanisms were also revealing. Technical problems, maintenance, and confidentiality of E-mail messaging were not found to be significant problems. Trends, value, and impact of E-mail use in health sciences institutions are also discussed. PMID- 8409810 TI - The empirical object of medical informatics. AB - Medical informatics is the science of information processing and the creation of information processing systems in medicine and health care delivery. Its methodological approach is based on the area specific applicability of a multidisciplinary theory of engineering and managing computerized information systems related to its empirical object. This paper gives a systemic view of the health care system, representing the empirical object of medical informatics. PMID- 8409811 TI - EM-PSYCH: a training program scheduling database. AB - EM-PSYCH is a scheduling database for training programs. It stores scheduling records and generates schedules and letters informing students, supervisors, and advisors of deadlines. Since EM-PSYCH was originally designed for a psychoanalytic institute, the institute model is presented as well as the database model to illustrate how the database program can be applied. EM-PSYCH is very user friendly and makes scheduling and sending reminders efficient and easy. It can handle any number of trainees and can be modified for any number of years or schedule interval. It can be easily adapted for psychiatric residencies, medical student education, psychology internships, and other training programs. PMID- 8409812 TI - Relationship of ST-segment elevation to eventual QRS loss in acute anterior wall myocardial infarction. AB - The position of electrocardiographic ST-segment elevation in acute myocardial infarction (MI) is related to the region of infarction, but the relationship between the initial ST-segment elevation and the eventual loss of QRS voltage remains uncertain. The authors examined body surface map data in 76 patients with acute anterior wall MI to clarify this relationship. The patients had no evidence of previous MI, no subsequent MI, and did not receive thrombolytic therapy or other acute interventions. Maps were recorded on admission to the hospital, every 24 hours during the hospital stay, and again at follow-up examinations 6-48 months after the index MI. The region of ST-segment elevation on the initial body surface map was compared to the region of developed voltage loss in the QRS complex as measured by the integral 0-30, 0-40, 0-80, and 30-80 ms body surface maps subsequently recorded 24 hours, 48 hours, and over 6 months after the initial body surface map. There was a relationship between the position of the ST segment elevation and the loss of the QRS segment voltages, which was pronounced early after acute MI and diminished months after the MI. The region of ST-segment elevation in the initial body surface map predicts the QRS loss. In the integral QRS0-30 map versus the initial ST-segment map the mean correlation coefficient was -0.54 +/- 0.33 with a median value of -0.67. The integral QRS0-40 map and ST segment map had a mean correlation coefficient of 0.60 +/- 0.29 and median of 0.66. The mean correlation coefficient is less for the integral QRS0-80 map and nonexistent in the integral QRS30-80 map. This method may be useful for monitoring acute interventions in acute MI. PMID- 8409813 TI - Comparative study of QRST values from body surface potential mapping, 12-lead ECGs, VCGs in detecting inferior myocardial infarction, and evaluating the severity of left ventricular wall motion abnormalities in simulated left bundle branch block. AB - The authors compared the ability of QRST time-integral values (QRST values) from body surface potential maps (BSPM), 12-lead electrocardiograms (ECGs), and Frank lead vectorcardiograms (VCGs) in diagnosing a prior inferior myocardial infarction (MI) in simulated left bundle branch block (LBBB). The study included 32 patients whose digitized ECGs were recorded simultaneously for BSPM, ECGs, and VCGs during normal sinus rhythm and during right ventricular pacing simulating LBBB (18 with and 14 without an inferior MI). QRST values were calculated in each lead point of ECGs. Data on 608 normal subjects were used as controls; mean +/- 2 SD was regarded as the normal range. The following parameters were derived: sigma DM, sigma DE, sigma DV, the sum of the differences between the normal mean QRST value, and the QRST value of a given patient in leads where the QRST value was less than the normal range ("-2 SD area") in BSPM, ECGs, and VCGs (Y lead). The correlation coefficients for sigma DM, sigma DE, and sigma DV between the two activation sequences were highly significant. Sensitivity and specificity were as follows: 89% and 93% for sigma DM > 100 mV.ms, 89% and 93% for sigma DE > 50 mV.ms, and 56% and 100% for sigma DV > 10 mV.ms, respectively. Although sigma DM, sigma DE, and sigma DV were significantly (P < .01) correlated with the asynergy index calculated from left ventriculograms, sigma DM showed the best correlation. QRST values from BSPM, ECGs, and VCGs provide information that is useful in detecting an inferior MI and in estimating the severity of left ventricular wall motion abnormalities in the setting of LBBB. Of the three parameters, BSPM showed the best correlation with the severity of left ventricular wall motion abnormalities. PMID- 8409814 TI - Improved exercise test accuracy using discriminant function analysis and "recovery ST slope". AB - The objective of the study was to optimize the accuracy of the exercise test for predicting the presence of significant angiographic coronary artery disease. A retrospective analysis of stored digital exercise electrocardiographic data on 147 men who had undergone exercise testing and cardiac catheterization was performed. With significant coronary artery disease defined as > or = 70% stenosis, 95 patients had one or more vessel(s) diseased. None were receiving digoxin, had a myocardial infarction or previous coronary artery bypass graft, or exhibited left bundle branch block, left ventricular hypertrophy, Q waves, or ST depression on their resting electrocardiogram. Analysis was performed using the authors' averaging and measurement software at rest and at each 30 seconds throughout the exercise and recovery in leads II, V2, and V5. Discriminant function analysis was used to analyze pretest variables, as well as hemodynamic and electrocardiographic changes and symptoms during exercise. A discriminant function score was developed and compared to other treadmill scores. The setting was a 1,000 bed Veterans Affairs Medical Center (Long Beach, CA). Discriminant function analysis chose age, smoking status, presenting chest pain characteristics, and lead V5 ST slope in recovery to have independent power for separating those with and without coronary artery disease. A discriminant function score using these four variables was used to form a receiver operating characteristics curve (and derive receiver operating characteristics curve areas) for comparison to other exercise test methods and scores: (discriminant function score = .81; slope 3.5 minutes into recovery in lead V5 = .73; traditional ST amplitude method = .72; ST60/HR index (amplitude of ST depression 60 ms after the J point/delta heart rate) = .66; traditional ST amplitude/HR index (traditional method/delta heart rate) = .75; Hollenberg score = .68; Hollenberg areas only = .66; and ST integral = .66. Receiver operating characteristics curve analysis revealed a trend for the discriminant function score to be superior to all other measurements and scores. Recovery ST slope in lead V5 performed as well as or better than all other electrocardiographic criteria or treadmill scores except for the authors' discriminant function score. PMID- 8409815 TI - Effect of autonomic blockade on digoxin-induced ECG changes at rest and during exercise in healthy subjects. AB - The effect of pharmacological autonomic blockade on digoxin-induced electrocardiographic changes at rest and during exercise was studied in nine healthy men. The subjects performed bicycle exercise tests on four occasions after: (1) intravenous injection of sodium chloride, (2) intravenous injection of propranolol and atropine, (3) oral ingestion of digoxin 0.5 mg daily for 2 weeks and intravenous injection of sodium chloride, and (4) oral ingestion of digoxin 0.5 mg daily for 2 weeks and intravenous injection of propranolol and atropine. The ST-T segment was significantly depressed after digoxin plus sodium chloride and digoxin plus propranolol and atropine in comparison with sodium chloride only and propranolol and atropine only. The digoxin-induced ST-T changes after sodium chloride and propranolol-atropine injections were not significantly different. Thus, judging from the results of this study, an effect mediated via the autonomic nervous system is an unlikely explanation of digitalis-induced ST-T changes. PMID- 8409816 TI - Electrophysiological abnormalities before and after surgery for atrial septal defect. AB - The authors evaluated the electrophysiological parameters, atrial-His interval, His-ventricular interval, Wenckebach cycle length, corrected sinus node recovery time (CSNRT), and sinoatrial conduction time (SACT) before and after operation in 28 children with atrial septal defect (ASD). Before operation, electrophysiological abnormalities were detected in 20 (79%) of the 28 patients. Preoperative comparison of the mean value in each parameter between the two age groups showed higher values in all parameters in the older group. In 17 patients who underwent operation, atrial-His interval, Wenckebach cycle length, CSNRT, and atrial effective refractory period (AERP) significantly decreased after operation; preoperatively abnormal Wenckebach cycle length, CSNRT, and AERP values were normalized in many of them. Thus, electrophysiological abnormalities were already present before operation in the ASD children and were severer in the older children, but were improved to some degree following operation. PMID- 8409817 TI - Minor recorder imperfections can cause artifacts in the RR interval spectrum derived from Holter recordings. AB - Spectral analysis of the heartbeat (RR) interval is a powerful tool for noninvasively assessing the autonomic nervous system. In addition, it may prove to be valuable in stratifying patients at risk for cardiac death. The authors report on a case in which spectral analysis of the RR interval exhibited harmonically related peaks within the physiological range that defied physiological explanation. Analysis showed that these peaks resulted from a subtle abnormality of the Holter recorder that was not apparent in the observed electrocardiogram. Since this type of abnormality can be produced by either minor damage or a manufacturing error in any brand of Holter recorder, the RR interval spectrum derived form a Holter electrocardiogram should always be critically examined for this particular artifact. PMID- 8409818 TI - Utility of transtelephonic ECG monitoring in evaluating discharges from a third generation implantable cardioverter defibrillator. AB - The authors report the case of a 60-year-old man with a third-generation implantable cardioverter defibrillator whose rate-sensing lead malfunction was diagnosed only after surface loop electrocardiographic (ECG) monitor recording. The stored R-R intervals suggested an irregular ventricular rhythm, but the loop ECG monitor demonstrated inappropriate shocks delivered during sinus rhythm. PMID- 8409819 TI - Neural network analysis is now commonplace in the medical literature. PMID- 8409820 TI - Proposal for an editorial policy for publications describing the use of artificial neural networks in electrocardiography. PMID- 8409821 TI - Fixed-interval performance and self-control in infants. AB - Twenty-six infants, 3 to 23 months old, were trained on fixed-interval schedules ranging from 10 s to 80 s. The operant response was touching an illuminated location on a touch-sensitive screen, and 20 s of cartoon presentation was the reinforcer. The subjects were also trained in a six-phase self-control procedure in which the critical phases involved choice between 20 s of cartoon available after a 0.5-s delay (impulsive choice) and 40 s of cartoon delayed for 40 s (self controlled choice). All the youngest children (3 to 5 months) showed long postreinforcement pauses on the fixed-interval schedule, with most intervals involving the emission of a single, reinforced, response, and all made self controlled choices. Older subjects (9 to 23 months) either produced the same pattern as the younger ones on the fixed-interval schedule (classified as pause sensitive subjects) or produced short pauses and higher steady response rates (classified as pause-insensitive subjects). All pause-sensitive subjects made self-controlled choices in the self-control condition, and all pause-insensitive subjects made impulsive ones. PMID- 8409822 TI - Tests of behavior momentum in simple and multiple schedules with rats and pigeons. AB - Four experiments examined the relationship between rate of reinforcement and resistance to change in rats' and pigeons' responses under simple and multiple schedules of reinforcement. In Experiment 1, 28 rats responded under either simple fixed-ratio, variable-ratio, fixed-interval, or variable-interval schedules; in Experiment 2, 3 pigeons responded under simple fixed-ratio schedules. Under each schedule, rate of reinforcement varied across four successive conditions. In Experiment 3, 14 rats responded under either a multiple fixed-ratio schedule or a multiple fixed-interval schedule, each with two components that differed in rate of reinforcement. In Experiment 4, 7 pigeons responded under either a multiple fixed-ratio or a multiple fixed-interval schedule, each with three components that also differed in rate of reinforcement. Under each condition of each experiment, resistance to change was studied by measuring schedule-controlled performance under conditions with prefeeding, response-independent food during the schedule or during timeouts that separated components of the multiple schedules, and by measuring behavior under extinction. There were no consistent differences between rats and pigeons. There was no direct relationship between rates of reinforcement and resistance to change when rates of reinforcement varied across successive conditions in the simple schedules. By comparison, in the multiple schedules there was a direct relationship between rates of reinforcement and resistance to change during most tests of resistance to change. The major exception was delivering response independent food during the schedule; this disrupted responding, but there was no direct relationship between rates of reinforcement and resistance to change in simple- or multiple-schedule contexts. The data suggest that rate of reinforcement determines resistance to change in multiple schedules, but that this relationship does not hold under simple schedules. PMID- 8409823 TI - Temporal control on interval schedules: what determines the postreinforcement pause? AB - On fixed-interval or response-initiated delay schedules of reinforcement, the average pause following food presentation is proportional to the interfood interval. Moreover, when a number of intervals of different durations occur in a programmed cyclic series, postreinforcement pauses track the changes in interval value. What controls the duration of postreinforcement pauses under these conditions? Staddon, Wynne, and Higa (1991), in their linear waiting model, propose control by the preceding interfood interval. Another possibility is that delay to reinforcement, signaled by a key peck and/or stimulus change, determines the subsequent pause. The experiments reported here examined the role of these two possible time markers by studying the performance of pigeons under a chained cyclic fixed-interval procedure. The data support the linear waiting model, but suggest that more than the immediately preceding interfood interval plays a role in temporal control. PMID- 8409824 TI - DRL interresponse-time distributions: quantification by peak deviation analysis. AB - Peak deviation analysis is a quantitative technique for characterizing interresponse-time distributions that result from training on differential reinforcement-of-low-rate schedules of reinforcement. It compares each rat's obtained interresponse-time distribution to the corresponding negative exponential distribution that would have occurred if the rat had emitted the same number of responses randomly in time, at the same rate. The comparison of the obtained distributions with corresponding negative exponential distributions provides the basis for computing three standardized metrics (burst ratio, peak location, and peak area) that quantitatively characterize the profile of the obtained interresponse-time distributions. In Experiment 1 peak deviation analysis quantitatively described the difference between the interresponse-time distributions of rats trained on variable-interval 300-s and differential reinforcement-of-low-rate 72-s schedules of reinforcement. In Experiment 2 peak deviation analysis differentiated between the effects of the psychomotor stimulant d-amphetamine, the anxiolytic compound chlordiazepoxide, and the antidepressant desipramine. The results suggest that peak deviation analysis of interresponse-time distributions may provide a useful behavioral assay system for characterizing the effects of drugs. PMID- 8409825 TI - The effects of SDRNFLRFamide and TNRNFLRFamide on the motor patterns of the stomatogastric ganglion of the crab Cancer borealis. AB - TNRNFLRFamide was isolated and sequenced from the stomatogastric nervous system of the crab Cancer borealis by reverse-phase high performance liquid chromatography followed by automated Edman degradation. An SDRNFLRFamide-like peptide that exactly co-migrated with SDRNFLRFamide was also observed. The effects of TNRNFLRFamide and SDRNFLRFamide on the gastric and pyloric rhythms of the stomatogastric nervous system of the crab Cancer borealis were studied. Both peptides activated pyloric rhythms in quiescent preparations in a dose-dependent manner with a threshold between 10(-11) and 10(-10) mol l-1. Both peptides increased the pyloric rhythm frequency of preparations showing moderate activity levels and had relatively little effect on preparations that showed strong pyloric rhythms prior to peptide application. Both peptides evoked gastric mill activity in preparations without existing gastric rhythms. The activation of the gastric rhythm is associated with activation of oscillatory properties in the dorsal gastric neurone. The induction of gastric rhythms by these peptides was accompanied by switches from pyloric-timed activity to gastric-timed activity by several stomatogastric ganglion neurones. Application of these peptides provides direct experimental control of circuit modification in the stomatogastric nervous system. PMID- 8409826 TI - Neuromuscular organization in the swimming system of the pteropod mollusc Clione limacina. AB - Swim motor neurons of the pteropod mollusc Clione limacina were identified by a combination of electrophysiological and morphological characteristics. Two types of motor neurons were found, including small motor neurons which are active during both slow and fast swimming and which innervated restricted fields of the ipsilateral wing. General excitor motor neurons have large cell bodies, innervate widespread fields and are recruited into activity for fast swimming. Small motor neurons monosynaptically innervate slow-twitch muscle cells, whereas general excitors monosynaptically innervate both slow-twitch and fast-twitch muscle cells. Activity in general excitors can centrally enhance that in small motor neurons because the neurons are electrically coupled. Neuromuscular recordings and lesion experiments indicate that a peripheral nerve network does not appear to play an important role in the spread of excitation throughout the muscle fields. PMID- 8409827 TI - Glucose concentration regulates freeze tolerance in the wood frog Rana sylvatica. AB - In spring, the lowest temperature during freezing that can be survived by wood frogs (Rana sylvatica) from southern Ohio is approximately -3 degrees C. We investigated whether the thermal limit of freeze tolerance in these frogs is regulated by tissue levels of glucose, a putative cryoprotectant that is distributed to tissues during freezing. Frogs receiving exogenous glucose injections prior to freezing showed dose-dependent increases in glucose within the heart, liver, skeletal muscle and blood. Tissue glucose concentrations were further elevated during freezing by the production of endogenous glucose. Most glucose-loaded frogs survived freezing to -5 degrees C, whereas all control (saline-injected) frogs succumbed. Further, we investigated some mechanisms by which glucose might function as a cryoprotectant in R. sylvatica. Organ dehydration, a normal, beneficial response that reduces freezing injury to tissues, occurred independently of tissue glucose concentrations. However, elevated glucose levels reduced both body ice content and in vivo erythrocyte injury. These results not only provided conclusive evidence for glucose's cryoprotective role in R. sylvatica, but also revealed that tissue glucose level is a critical determinant of freeze tolerance capacity in this species. PMID- 8409828 TI - Extracellular ATP selectively modulates a high-voltage-activated calcium conductance in salivary gland cells of the leech Haementeria ghilianii. PMID- 8409829 TI - Thoracic connections between crayfish giant fibres and motor giant neurones reverse abdominal pattern. PMID- 8409830 TI - Autoimmune endocrinopathies. PMID- 8409831 TI - Recombinant thyroid autoantigens: the keys to the pathogenesis of autoimmune thyroid disease. PMID- 8409832 TI - Islet cell autoimmunity. PMID- 8409833 TI - The association of autoantibodies directed against ovarian antigens in human disease: a clinical review. PMID- 8409834 TI - Autoimmune disease of the adrenal cortex, pituitary, parathyroid glands and gastric mucosa. PMID- 8409835 TI - Rationale and efficacy of medical therapy for gastrooesophageal reflux disease. PMID- 8409836 TI - Diacylglycerol/protein kinase C signalling: a mechanism for insulin resistance? AB - It is proposed that an intracellular cycle exists to limit or terminate the insulin signal. The cycle involves increased synthesis of sn-1,2-diacylglycerol (DAG) in response to insulin. The DAG activates protein kinase C (PKC) which phosphorylates glycogen synthase either directly or through other protein kinases to render it inactive. Protein kinase C may also inhibit the insulin receptor by phosphorylation of receptor serine residues. Insulin resistance could then arise as a consequence of a persistent increase in DAG levels. Such an increase could occur in three different ways. Chronic hyperinsulinaemia could increase DAG levels by de-novo synthesis from phosphatidic acid, by hydrolysis of phosphatidylcholine, or by hydrolysis of glycosyl-phosphatidylinositol; DAG is also formed by hydrolysis of phosphatidylinositol 4,5-biphosphate (PIP2). This reaction, known as the 'PI response,' may be the connection between hypertension and insulin resistance. A third mechanism for an increase in DAG involves neural abnormalities. Thus, muscle denervation in the rat is characterized both by a profound insulin resistance and a large increase in DAG. It is possible that a similar increase occurs in humans and may explain the association between denervation, inactivity, and insulin resistance. PMID- 8409837 TI - Sonography of the Achilles tendon in hypercholesterolaemia. AB - OBJECTIVES: Tendon xanthomas cause thickening of the tendon and are an important sign in monogenic familial hypercholesterolaemia (FH). The aim of our study was to investigate the usefulness of achilles tendon sonography in detecting FH patients. Special attention was paid to structural abnormalities of the achilles tendon. DESIGN: A clinical study with methodological testing. SETTING: Patients suspected of having FH were sent to the out-patient Department of Medicine from other departments of Turku University Central Hospital and from primary care units. The patients were studied by high-frequency ultrasound before more exact typing of the lipid disorder. An additional study of normolipidaemic volunteers and a phantom study were also carried out. SUBJECTS: Forty FH patients, 51 non-FH hypercholesterolaemia patients and 41 normolipidaemic volunteers were included in the study. MAIN OUTCOME OF MEASURES: The thickness of the tendon was measured and the tendon structure and its echogenicity were recorded. RESULTS: Twenty-five out of 40 (63%) FH patients had distinctly thickened tendons (men more than 10 mm, women more than 9 mm). Thirty-six (90%) had a typical structural alteration of low or mixed echogenicity of the tendon. Three non-FH patients were found to have xanthomas on sonography. CONCLUSIONS: We conclude that ultrasonography is a sensitive method of detecting xanthomas that reveals the altered tendon structure even in xanthomatous tendons of normal thickness. PMID- 8409838 TI - Behaviour therapy versus doctor's anti-smoking advice in diabetic patients. AB - OBJECTIVES: To evaluate the efficacy of a structured behaviour therapy programme on smoking cessation in diabetic patients. DESIGN: Prospective, randomized, controlled intervention study. SETTING: University out-patient diabetes clinic. SUBJECTS: A total of 794 consecutive insulin-treated smoking diabetic patients were invited to participate in a smoking cessation programme. Eighty-nine patients agreed to participate and were randomized in two groups. INTERVENTIONS: Forty-four patients were randomized to a structured extensive behaviour therapy anti-smoking intervention and 45 patients to a control group that received a single unstructured anti-smoking advice session given by a physician. MAIN OUTCOME MEASURES: After 6 months, nine patients were confirmed not to be smoking (i.e. urine cotinine concentration below 20 ng ml-1, 2 [5%] in the behaviour therapy intervention group and 7 [16%] in the control group. CONCLUSIONS: In diabetic patients an extensive behaviour therapy intervention for smoking cessation is no more successful than an unstructured physician's advice. PMID- 8409839 TI - Hyperlipidaemia--management and views amongst physicians in general practice, in occupational health care and in internal medicine. AB - OBJECTIVES: To study aspects of management and views about hyperlipidaemia and its treatment amongst different categories of physicians. DESIGN: A postal questionnaire was sent to selected physicians. Reminders were used. SETTING: The study included physicians in primary health care (PHCPs), occupational health care (OHCPs), and in departments of internal medicine in hospitals (IMPHs). SUBJECTS: All 146 PHCPs in the south-western region of Stockholm, 147 OHCPs, randomly selected from the directory of occupational health care offices, and all 157 IMPHs in six hospitals, in the greater Stockholm area, were offered the questionnaire. MAIN OUTCOME MEASURES: Differences in views, attitudes and treatment policy amongst different categories of physicians. RESULTS: Response rate was 70% (61-81%). Intervention was initiated by the OHCPs at lower levels of cholesterol than by the other two groups, with diet (P < 0.001) and with drug intervention as primary (P < 0.01) and secondary prevention (P < 0.005). When other cardiovascular risk factors were present, intervention was started earlier by the OHCPs only in comparison with the PHCPs (P < 0.01). Knowledge of one's own cholesterol level was more common amongst the OHCPs than in the other two groups (P < 0.001). Patient follow-up was the same in all groups. CONCLUSIONS: Disagreement amongst physicians about hyperlipidaemia leads to conflicting information being given to patients. It is important to be aware of these differences when trying to reach a consensus on this topic. Corresponding knowledge and attitudes amongst members of the public should also be studied. PMID- 8409840 TI - Hypoglycaemia caused by atypical insulin antibodies in a patient with benign monoclonal gammopathy. AB - We describe a 48-year-old woman with recurrent severe hypoglycaemia apparently caused by a paraprotein with insulin-binding capacity. Very high fasting values were found for serum insulin (170 and > 250 mU l-1) as well as for proinsulin 125 pmol l-1 and an insulinoma was suspected. Hypoglycaemia developed after an oral glucose tolerance (OGTT) test but not during fasting for 48 h. Free insulin and C peptide were normal during OGTT whereas serum insulin was very high. 125I-insulin binding to serum, determined with a polyethylene glycol (PEG) precipitation method was high (40%), and equally high after addition of 1.7 x 10(-5) mol l-1 cold insulin to estimate non-specific binding. By adding very high concentrations of cold insulin, displacement of 125I-insulin bound to serum was found (50% displacement at 4 x 10(-5) mol l-1). No immunoglobulin G (IgG) insulin antibodies were detected by radioimmunoelectrophoresis. On agarose electrophoresis a small paraprotein (4 g l-1) in the gamma-globulin fraction was detected. 125I-insulin binding to this paraprotein was demonstrated. We conclude that if insulin autoantibodies are suspected as a cause of hypoglycaemia screening for insulin antibodies should always be done with a PEG-precipitation method. PMID- 8409841 TI - Central hypothyroidism presenting with pericardial and pleural effusions. PMID- 8409842 TI - Syndrome X. PMID- 8409843 TI - Accelerated coronary artery disease after heart transplantation: the role of enhanced platelet aggregation and thrombosis. PMID- 8409844 TI - Pathophysiological mechanisms operating in restenosis following coronary angioplasty. PMID- 8409845 TI - Analysis of T-helper and T-suppressor lymphocyte subsets in patients with chronic hepatitis C virus infection. PMID- 8409846 TI - The time course of perceptual and conceptual contributions to word fragment completion priming. AB - Two experiments examined the time course of the availability of perceptual and conceptual information in priming on the word fragment completion test. Subjects encoded primes as either visual words, auditory words, or pictures. In Experiment 1, word fragments were exposed for either 500 ms, 1 s, 5 s, or 12 s. Only the visual words produced priming at the 500-ms and 1-s exposure times. In Experiment 2, subjects were allowed up to 20 s to solve each fragment; response latencies were recorded and cumulative response curves were generated. Visually primed fragments were solved at a faster rate than either auditorily or pictorially primed fragments. The results suggest that although conceptual processing can contribute to word fragment priming, perceptual processes are recruited earlier and at a faster rate. PMID- 8409847 TI - Discourse models, pronoun resolution, and the implicit causality of verbs. AB - Some interpersonal verbs, such as admire and amaze, describe an action or property of one person (the reactor) that is necessarily a response to an action or property of another (the initiator). We hypothesized that these verbs make the initiator relatively more accessible in a comprehender's discourse model and that this change in relative accessibility aids identification of the referent of a pronoun in a subsequent because clause. We predicted that, as a result, subjects would be faster to recognize a character's name after a because clause that uses a pronoun to refer to that character than after one that refers to some other character. Four experiments confirmed this prediction. Three further experiments demonstrated the importance of the verb's causal structure and of the presence of the connective because to this result. PMID- 8409848 TI - Connecting goals and actions during reading. AB - Three experiments showed that reading about a character's actions can reactivate a goal of the character stated earlier in the passage and backgrounded by intervening material. Subjects were slower to read a line describing an action that was inconsistent with a goal of the protagonist than they were to read about an action that was consistent with the goal, even though both lines were locally coherent. Goals were reactivated even when the intervening material did not describe attempts to achieve the goal (Experiment 2) and when the intervening material described another goal of the protagonist (Experiment 3). The results suggest that reading a sentence can reactivate relevant information from earlier in the text, even when the sentence is coherent with its immediate context and the reactivated information has been backgrounded by several lines of unrelated text. PMID- 8409849 TI - Reduction in alphabet priming with delay and degradation. AB - Phonologically ambiguous Serbo-Croatian words are named more slowly than their phonologically unique partners. This difference is reduced by nonword primes containing consonants unique to one or the other alphabet. In 2 experiments we investigated the hypothesis that alphabet priming is the inhibition of unique and ambiguous letter units of one alphabet by the unique letter units of the other alphabet. In Experiment 1, ambiguous and unique words followed alphabet-specific nonwords at lags between 100 ms and 1,550 ms. The ambiguous-unique difference increased from 1 ms to 45 ms, consistent with a relaxing inhibitory process. In Experiment 2 we compared priming of ambiguous words with and without visual noise. Priming was less for noisy than for intact stimuli, as would be expected if noise slows processing and if the inhibition responsible for priming weakens further during the additional processing time. PMID- 8409850 TI - Working-memory capacity as long-term memory activation: an individual-differences approach. AB - One explanation of the correlation often observed between working-memory span scores and reading comprehension is that individuals differ in level of activation available for long-term memory units. Two experiments used the fan manipulation to test this idea. In Experiment 1, high- and low-working-memory Ss learned a set of unrelated sentences varying in the number of shared concepts (fan) and then performed speeded recognition for those sentences. Low-working memory Ss showed a larger increase in recognition time as fan increased. When the slope of the fan effect was partialed out of the relationship between working memory span and verbal abilities, the relationship was reduced to nonsignificance. In Experiment 2, Ss learned thematically related sentences that varied in fan. Low-span Ss showed the positive fan effect typically found with thematically unrelated sentences, whereas high-span Ss showed a negative fan effect. The results are discussed in terms of a general capacity theory. PMID- 8409851 TI - The many facets of repetition: a cued-recall and event-related potential analysis of repeating words in same versus different sentence contexts. AB - Event-related potential (ERP) and cued-recall performance were used to investigate the influence of (a) context, (b) repeating a word's meaning to word repetition priming, and (c) repetition on the ERP difference related to memory (Dm). Sentences ended with either nonhomographs or homographs. For nonhomographs, either the sentence context, the final word, both, or neither were repeated. Homographs were repeated in their original context or in new sentences that biased the same or an alternative meaning. Large repetition effects were found for all words repeated in their original contexts; in contrast, changing contexts led to no repetition effects whether the meaning of the repeated words was preserved or not. These results favor an episodic contribution to word repetition priming and suggest a common process for Dm and repetition. PMID- 8409852 TI - Reduced feedback frequency enhances generalized motor program learning but not parameterization learning. AB - The purpose of the study was to examine the effects of a reduced feedback frequency on the learning of generalized motor programs and movement parameterization. Subjects practiced three movement patterns with the same relative timing and the same relative amplitude, but with varied movement time (Experiment 1) or varied movement amplitude (Experiment 2). KR was given either on 100% or 63% of the trials, with learning being assessed by retention and transfer tests. In both experiments, reduced KR frequency enhanced GMP learning but generally degraded parameter learning. These data provide converging evidence for the dissociation of the program and parameterization processes postulated in GMP theory. PMID- 8409853 TI - Determinants of diagnostic hypothesis generation: effects of information, base rates, and experience. AB - Physicians generated diagnostic hypotheses for case histories for which 2 types of diagnoses were plausible, with one having a higher population base rate but less severe clinical consequences than the other. The number of clinical and background symptoms pointing towards the 2 diagnoses was factorially manipulated. The order and frequency with which physicians generated hypotheses varied with the amount of relevant clinical and background information and as a function of population incidence rates, with little evidence of base rate neglect. Availability of a hypothesis, made possible by diagnosis of a similar case before, also made doctors generate this diagnosis earlier and more frequently. Physicians' experience affected hypothesis generation solely by increasing the availability of similar cases. The results are consistent with the use of similarity-based hypothesis generation processes that operate on memory for prior cases. PMID- 8409854 TI - Implicit and explicit memory for novel visual objects: structure and function. AB - Six experiments compared the effects of structural and functional encoding tasks on implicit and explicit memory for novel objects. Implicit memory was assessed with a possible-impossible object decision test, and explicit memory was assessed with a yes-no recognition test. Results revealed that recognition memory was higher after functional than after structural encoding tasks, whereas priming effects on the object decision test were unaffected by the same manipulations. The priming effects that were observed after functional encoding tasks could be attributed to structural analyses that are carried out in the course of making judgments about functional properties of novel objects. Results are consistent with the hypothesis that implicit memory for novel objects depends on a presemantic structural description system that can operate independently of episodic memory. PMID- 8409855 TI - An empirical and meta-analytic evaluation of the phoneme identification task. AB - Recent studies that used Ganong's (1980) identification task have produced discrepant results. The present study sought to resolve these discrepancies by examining the influence of methodological factors on phoneme identification and differences in data analysis techniques. Three factors were examined across 2 experiments: position of target phoneme, phonetic contrast, and 2 task conditions in which stimulus quality (S/N ratio) or cognitive load varied. A meta-analysis was then performed on the results from all identification studies, including the present one, in an effort to obtain additional insight on factors that influence the task. The experiments and meta-analysis identified the importance of several methodological factors in affecting identification, most notably position of the target phoneme. PMID- 8409856 TI - Haptic exploration in the presence of vision. AB - Two experiments addressed initiation of haptic exploration to encode object properties when vision is present. Ss compared pairs of objects on designated properties, using only vision or with touch permitted. Tough initiation, reach, contact, and visual responses were timed. With difficult material judgments, tough occurred frequently and was initiated faster than the time to respond by vision alone. With geometric judgments, touch was rarely used and then was initiated at the typical time for a visual response. Imposing a visual preview before allowing touch did not reduce the incidence of touch but did speed its initiation. Results support a model in which preliminary visual processing quickly initiates haptic exploration for material judgments that are visually or semantically difficult. PMID- 8409857 TI - Strategic control of processing in word recognition. AB - Strategic control of word recognition in a lexical decision task was examined by manipulating the similarity of nonword foils to real words (nonword lexicality). Overall correct reaction times to words and the advantage of high- over low frequency words were greater when nonword foils were more wordlike. This was true for both illegal (BTESE) versus legal (DEEST) nonword foils and legal nonword versus pseudohomophone (BEEST) foils. The same pattern of results was replicated in a 2nd experiment in which the word targets were always irregular (e.g., HAVE). A 3rd experiment demonstrated a large frequency blocking effect for low-frequency words, given pseudohomophone foils. The results are applied to pathway selection and random-walk frame-works. For both framework, canonical models are developed, which characterize qualitative predictions of broad classes of models within that framework. We argue for a pluralistic approach to theory development that moves from lower to higher order isomorphisms between data and theory. PMID- 8409858 TI - Asymmetries in visual search for conjunctive targets. AB - Asymmetry is demonstrated between conjunctive targets in visual search with no detectable asymmetries between the individual features that compose these targets. Experiment 1 demonstrated this phenomenon for targets composed of color and shape. Experiment 2 and 4 demonstrate this asymmetry for targets composed of size and orientation and for targets composed of contrast level and orientation, respectively. Experiment 3 demonstrates that search rate of individual features cannot predict search rate for conjunctive targets. These results demonstrate the need for 2 levels of representations: one of features and one of conjunction of features. A model related to the modified feature integration theory is proposed to account for these results. The proposed model and other models of visual search are discussed. PMID- 8409859 TI - Visual search times assessed without reaction times: a new method and an application to aging. AB - We consider several ways in which the interpretation of reaction time (RT) data might confound differences in visual search rates with non-search-related factors. To determine whether estimates of search rates for groups differing in age suffered this problem, we compared estimates provided by the RT method with those obtained using a forced-choice method with limited-duration stimuli. The forced-choice method provided faster estimates of search rates. The effects of age, the variable in which we were interested, were comparable, but the difference between results obtained using the two methods suggests the need for caution in using the RT method. We discuss how the forced-choice method can be used, under appropriate circumstances, to provide an independent test of whether subjects are carrying out serial searches and, if they are, to provide search rate estimates even from data obtained using only a single array size. PMID- 8409860 TI - Time courses in the negative and positive repetition effects. AB - Time courses of the negative repetition effect (NRE), a poorer detection of the target in noise-same-as-target than in noise-alternative-target displays, and its opposite, the positive repetition effect (PRE), were examined. Experiment 1 showed that displays in which a low-contrast target was present with a high contrast noise produced a larger NRE than did displays in which a contrast relationship between items was reversed. A negative contrast repetition effect (NCRE) was also found that was comparable to the NRE. Experiment 2 showed that dimensional Korean letters (e.g., [symbol: see text]) whose configural differences were apparent in orientation produced the largest PRE at a 0-ms stimulus onset asynchrony, whereas featural Korean letters (e.g., [symbol: see text]) that differed in the number of elements yielded the largest NRE when a noise letter preceded a target letter by 50 ms. Experiments 3A and 3B indicated that the NCRE may arise from spatial attention. PMID- 8409861 TI - Interference effects in the Stroop and Simon paradigms. AB - The possibility that Stroop and Simon effects reflect the same cognitive processes was tested in 3 experiments with 62 adult Ss. The words LEFT and RIGHT were shown left and right of screen center. Similar levels of interference were found for a Simon task (keypress to meaning of word) and a spatial Stroop task (name its location). A reverse Simon task (keypress to word location) showed minimal interference from the irrelevant word, but in a reverse spatial Stroop task (read the word aloud) interference from the irrelevant location was sizable. This poses difficulty for translational accounts of Stroop interference. Presenting LEFT and RIGHT laterally balanced by a color-name foil yielded a Simon effect inconsistent with an orienting response hypothesis. With bilateral stimuli in a reverse Simon task, interference was marked, supporting an integrative account of Stroop and Simon effects in terms of cognitive activation and the salience of irrelevant stimuli. PMID- 8409862 TI - Information transfer in iconic memory experiments. AB - To report letters from briefly exposed letter arrays, subjects must transfer information from a rapidly decaying trace (iconic memory) to more durable storage. In a partial-report paradigm, we systematically varied the proportion (P) of trials with a long cue delay relative to a short cue delay. Practiced subjects used the same transfer strategy independent of P. Data from a partial report-plus-masking experiment were used to construct a computational model that accurately predicted partial- and whole-report performance with and without masks. Assumptions: Prior to a cue, subjects attend primarily to the middle row of a three-row display, resulting in nonselective transfer. After the cue, they attend only to the cued row. Transfer rate is the product of iconic legibility (which depends on time and retinal location) and attention allocation (which shifts after a cue). Cumulative transfer is limited by the capacity of durable storage. PMID- 8409863 TI - Frames of reference and distinctive figural characteristics affect shape perception. AB - In 4 experiments it was found that global frameworks and local distinctive figural characteristics influenced the perception of shape and of pointing. In Experiments 1 and 2, Ss were asked to mark the center of the middle figure in arrays of 3 aligned figures (either triangles or squares). Displacements of the center indicated a perceived deformation of the shapes. In Experiment 3, Ss were asked to adjust the height of triangles in arrays similar to those in Experiment 1. Height adjustments made along the axis of apparent pointing resulted in extents that were shorter than the height necessary to form equilateral triangles. In Experiment 4, stimuli were isosceles triangles in which the apparent distortion had been nulled; however, Ss continued to perceive them as pointing in a direction consistent with their alignment, and hence shape distortion cannot be the cause of pointing. PMID- 8409864 TI - Time course of object identification: evidence for a global-to-local contingency. AB - A time course contingency is the modification of later phases of object recognition contingent upon stimulus information extracted earlier in processing. It can increase the efficiency of later processing and reduce computational burdens. This idea was instantiated within a global-to-local model and supported in 4 integration priming experiments, in which primes and target objects were presented briefly and then masked. In Experiment 1-3, global and coarse-grained common-feature primes presented early in processing facilitated discriminations between similarly shaped objects, even though they provided no discrimination relevant information. In Experiment 4, global primes were more effective than local primes early in processing, whereas local primes were more effective than global primes late in processing. PMID- 8409865 TI - Color improves object recognition in normal and low vision. AB - Does color improve object recognition? If so, is the improvement greater for images with low spatial resolution in which there is less shape information? Do people with low visual acuity benefit more from color? Three experiments measured reaction time (RT) and accuracy for naming food objects displayed in 4 types of images: gray scale or color, and high or low spatial resolution (produced by blur). Normally sighted Ss had faster RTs with color, but the improvement was not significantly greater for images with low spatial resolution. Low vision subjects were also faster with color, but the difference did not depend significantly on acuity. In 2 additional experiments, it was found that the faster RTs for color stimuli were related to objects' prototypicality but not to their color diagnosticity. It was concluded that color does improve object recognition, and the mechanism is probably sensory rather than cognitive in origin. PMID- 8409866 TI - Electrocutaneous spatial integration at suprathreshold levels: an additive neural model. AB - The effect of cathode size on estimates of the magnitude of electrocutaneous stimulation were studied. The 24 Ss made magnitude estimates for combinations of 6 cathode sizes (2.5-30 mm diameter) and 5 currents (1.5-3.3 times the threshold). Estimates for small cathodes were consistently smaller but grew more rapidly than those for intermediate and large cathodes. From the results, contours were constructed showing how area and current trade off to maintain apparent intensity. These area-current contours suggest that supersummation prevails at low estimation levels and complete summation tasks place at high levels. An additive neural model is proposed to account for the results and is also applied to other somatosensory systems. PMID- 8409867 TI - Bronchitis. PMID- 8409868 TI - Osteopathy vs chiropractic. PMID- 8409869 TI - Colonoscopy. PMID- 8409870 TI - Colonoscopy. PMID- 8409871 TI - Hypothyroidism. PMID- 8409872 TI - Screening and brief intervention for alcohol disorders. PMID- 8409873 TI - Variation in physicians' recommendations about revisit interval for three common conditions. AB - BACKGROUND: The appropriate revisit interval for most conditions is uncertain. This survey was done to gather information about physicians' recommendations on revisit intervals for three common conditions. METHODS: Data were gathered in a mailed survey of 116 primary care physicians in the University of California at San Francisco Collaborative Research Network. Physicians were given descriptions of three hypothetical patients, one with diabetes mellitus, one with angina, and one with hypertension, and were asked when they would recommend a follow-up visit for the condition. RESULTS: There were great variations in physicians' recommendations about revisit intervals for each hypothetical patient. Internists were significantly more likely than family physicians to recommend a longer revisit interval for the hypothetical patient with diabetes mellitus; a similar revisit pattern was found for the patient with hypertension. There were no significant associations with recommended revisit interval and many other physician characteristics. CONCLUSIONS: As interest in containing the cost and improving the efficiency of medical care increases, knowing how often patients ought to be seen will be a topic of increasing importance. A rational, information-based approach to the choice of revisit interval for common conditions could yield substantial savings in medical care costs. The existence of great variation in recommended revisit interval suggests that physicians are uncertain about what interval is best. PMID- 8409874 TI - Patients' reactions to physician use of a computerized medical record system during clinical encounters. AB - BACKGROUND: As physicians begin to use computer technology in front of patients during clinical encounters, concern has been raised that such computer use may exert a dehumanizing effect on the physician-patient relationship. To investigate this concern, we measured patient reactions to physician use of a computerized medical record system during clinical encounters. METHODS: Adult patients who presented for clinical care were randomized into three groups. With the first group, the physician used a standard paper-and-pencil charting system during the encounter. With the second group, the physician used a computerized medical record system with keyboard input. With the third group, the physician used the computerized medical record system with voice input. Patient reactions were measured with a questionnaire that the patients completed after the clinical encounter. RESULTS: For most components of the physician-patient relationship studied in this report, questionnaire scores did not differ significantly among the three study groups. Patients in the voice input group rated physician explanations of patient problems significantly higher than patients in the other two groups. There was a trend for patient confidence in the physician to be higher in the keyboard input group. Although measured encounter durations were significantly shorter in the computer groups, there were no differences in patient satisfaction with encounter duration among the three groups. CONCLUSIONS: Physician use of computers during clinical encounters was not associated with a decline in the perceived quality of the physician-patient relationship. PMID- 8409875 TI - A comparison of after-hours telephone calls concerning ambulatory and nursing home patients. AB - BACKGROUND: This study documents the frequency and nature of after-hours telephone calls to a university-based family practice, with special attention to those calls from or about nursing home patients. METHODS: All after-hours telephone calls to a free-standing family practice training program that were made during the 6 months between July 1991 and January 1992 were recorded and classified. RESULTS: Of the 821 calls recorded, 81 included telephone calls from nursing home patients. Nursing home patients, who constitute about 1% of the practice, were responsible for 10% of the calls (P < .001). Nursing home patient calls were more likely to occur on weekends (P = .013) and were more likely to be for physician notification purposes (P < .001). CONCLUSIONS: Nursing home patients generate a disproportionately large number of after-hours calls. These calls are more likely to occur on weekends and less likely to require physician action. This is a considerable hidden practice burden that needs to be taken into account when planning practice coverage. PMID- 8409876 TI - Screening for proteinuria in patients with hypertension or diabetes mellitus. AB - BACKGROUND: Proteinuria is an early indication of renal disease. This study was conducted to evaluate the usefulness of dipstick urinalysis in patients with chronic diseases including hypertension and diabetes mellitus. METHODS: At a university family practice center, patients without urinary tract disorders underwent dipstick urinalysis. RESULTS: Of the 796 patients evaluated, increased proteinuria, possibly indicating early renal disease, was detected in 4% of healthy patients, 16% of patients with hypertension, 29% of patients with diabetes, and 53% of patients with both hypertension and diabetes. A higher incidence of proteinuria was found among African American patients with hypertension or diabetes or both than among white patients. CONCLUSIONS: Regular dipstick evaluation for proteinuria may be indicated in patients with hypertension or diabetes mellitus or both, particularly African American patients with these disorders. PMID- 8409877 TI - Professional satisfaction and dissatisfaction of family physicians. AB - BACKGROUND: Physicians' satisfaction with their professional life influences the quality of patient care they provide and helps to determine the number and type of students attracted to the various fields of medicine. In this study, we sought to delineate areas of satisfaction and dissatisfaction among family physicians. METHODS: A self-administered questionnaire was sent to all physicians in the state of Pennsylvania who were included in the 1990 directory of the American Board of Family Practice (N = 1944). RESULTS: Completed questionnaires were received from 1066 family physicians in full-time practice. Sixty-five percent were satisfied with their professional lives. Patient relationships, a sense of clinical competence, and their relationships with their partners were among the most satisfying aspects of practice for all family physicians. Problems identified included regulations by third-party payers and government agencies and the large amount of paperwork encountered in practice. There were significant (P < .001) differences in satisfaction between physicians in different practice arrangements. Significant differences between practice types were also found in the degree of dissatisfaction with third-party payers and government agencies, paperwork, isolation from other physicians, and the threat of a malpractice suit. CONCLUSIONS: Almost two thirds of family physicians are satisfied with their general professional lives. Conversely, one third are not. Clear areas of satisfaction and dissatisfaction have been defined for family physicians in general as well as for family physicians in various practice environments. This information may be useful in the development of policy to structure a medical system that meets the needs of both patients and physicians. PMID- 8409878 TI - Smokeless tobacco cessation: report of a preliminary trial using nicotine chewing gum. AB - BACKGROUND: Smokeless tobacco use is a major public health hazard whose incidence is increasing, particularly among male adolescents. Little research has been done on cessation programs designed to assist smokeless tobacco users in ending their habit. There have been no studies on the use of nicotine polacrilex chewing gum as an adjunct to cessation. METHODS: Fourteen of 88 male smokeless tobacco users in a professional baseball organization enrolled in a cessation program and were followed for up to 12 months. The program consisted of two support group sessions at the spring training camp followed by adjunctive use of nicotine polacrilex chewing gum during the baseball season as monitored by the athletic trainers. RESULTS: At 2 to 4 months, only 3 of 14 participants were completely abstinent from smokeless tobacco. Follow-up data at 6 to 12 months revealed that only one participant was abstinent. The 14 ballplayers experienced various side effects of nicotine chewing gum: bad taste (6), nausea (4), headache (4), jaw discomfort (3), and dizziness (1). Despite these side effects, 11 of the 14 participants replied that they would recommend the gum to others trying to quit. Most participants (10) felt that quitting the smokeless tobacco habit was "very difficult." CONCLUSIONS: We conclude that nicotine chewing gum as an adjunct to smokeless tobacco cessation had limited effectiveness. Further study on smokeless tobacco cessation methods is needed. PMID- 8409879 TI - Screening for gestational diabetes mellitus: a critical review. AB - Gestational diabetes mellitus (GDM) occurs in 1% to 3% of pregnant women. Generally, the clinical focus in these cases is on intermediate outcomes such as macrosomia, hypoglycemia, or hypocalcemia. Only macrosomia is consistently associated with gestational diabetes, yet the risks of macrosomia such as shoulder dystocia and birth injury are highly variable. The screening test and the reference standard, the oral glucose tolerance test, are problematic in that there are no standardized testing procedures or definitive criteria for diagnostic interpretation and poor reproducibility of test results. There have been no methodologically sound randomized controlled trials of therapy for GDM. Studies that attempted randomization show, however, that therapy reduces the incidence of macrosomia, which is an intermediate outcome. A critical review of the literature revealed that there is insufficient evidence to justify routine screening for gestational diabetes. A reassessment of the relation between maternal glucose levels in pregnancy and neonatal outcomes is needed to determine if there are correctable adverse outcomes. In the meantime, management should be based on careful assessment of each individual pregnancy. PMID- 8409880 TI - Hillary's Rx. PMID- 8409881 TI - Tobacco and pharmacies. PMID- 8409882 TI - Informed consent. PMID- 8409883 TI - "Back mouse". PMID- 8409884 TI - Physicians, patients, and third parties: everybody's talking but is anybody listening? PMID- 8409885 TI - Prejudices of a local medical doctor. PMID- 8409886 TI - Prevalence of depressive symptoms in primary care. AB - BACKGROUND: Depression is one of the most common medical disorders seen in primary care practice. The purpose of this study was to estimate the prevalence of depressive symptoms in primary care patients across the United States, and to describe patient characteristics that may be associated with an increased likelihood of those symptoms. METHODS: Survey data were obtained from a sample of 75,858 patients who visited one of 765 participating primary care physicians for any reason from February 1991 to September 1991. The outcome measurement used was the index score for presence of depressive symptoms on the Zung Self-rating Depression Scale. RESULTS: The overall prevalence of clinically significant depressive symptoms was found to be 20.9%, but the percentage of patients citing depression as a reason for visit (1.2%) was markedly lower. Patients who perceived their health as poor were more likely to have severe depressive symptoms than patients who perceived their health as excellent. Women, those in older age groups, and those with lower levels of education were more likely to have clinically significant depressive symptoms than men, those in younger age groups, and those with higher levels of education. When classified by marital status within each sex, married men and women were the least likely to have clinically significant depressive symptoms. CONCLUSIONS: Clinically significant depressive symptoms are highly prevalent in primary care patients; however, depression is an infrequent patient complaint. There are certain patient characteristics that may cue the physician to consider depression in the differential diagnosis, particularly the patient's self-perception of his or her overall health status. In addition, certain other subsets of patients are at increased risk of depression, such as women, those in older age groups, and those of lower socioeconomic status. PMID- 8409887 TI - Clinical evaluation of ankle inversion injuries in family practice offices. AB - BACKGROUND: The use of radiography in evaluating inversion ankle injuries remains high despite several studies suggesting that x-ray examination should be limited to patients meeting certain clinical criteria. These studies were all done in emergency departments. The present study examined detection of ankle fractures by clinical evaluation alone in private family practice offices. METHODS: Twelve physicians in three family practice offices participated. Check-off forms were developed to record clinical data. The physicians all attended a session to standardize terminology. The physicians then evaluated 94 consecutive patients with inversion ankle injuries. RESULTS: Eight fractures were detected by radiography, five of which had not been suspected on clinical examination (5.9% false-negative rate). Only one fracture required treatment different from that for a sprain. Tenderness on the dorsum of the foot, impaired weight-bearing ability, recentness of injury (less than 12 hours earlier), and presence of additional injuries were significantly associated with a fracture. Unlike several previous studies, swelling was not associated with fractures. If radiography had been limited to patients presenting with inability to bear weight fully or tenderness on the dorsum of the foot, none of the fractures would have been missed, and the use of radiography would have been reduced from 90% to 61%. CONCLUSIONS: The fracture rate in these family practice offices is lower than that reported in most emergency department studies. It is important that family physicians order radiographs judiciously rather than routinely for patients with inversion ankle injuries. The clinical criteria reported here are likely to reduce unnecessary ordering of radiographs and are compatible with recently published, prospectively validated rules for acute ankle injury in an emergency department setting. PMID- 8409888 TI - Access to medical care among the elderly in rural northeastern Ohio. AB - BACKGROUND: Conventional wisdom holds that the elderly living in rural areas suffer poor health and have limited access to health care compared with their urban peers. The relation between poor health and limited access, however, has yet to be adequately defined. METHODS: We conducted a telephone survey of 1000 elderly persons living in four rural northeastern Ohio counties using a proportional random-digit dialing method. RESULTS: Many rural elderly respondents appeared to suffer poor health and have limited access to medical care. However, a detailed analysis revealed that poor health and limited access were more perceptual than actual. CONCLUSIONS: Elderly persons living independently in rural northeast Ohio have much better health and access to care than suggested by the literature. PMID- 8409889 TI - Factors influencing mammogram ordering at the time of the office visit. AB - BACKGROUND: Breast cancer is the second most common cause of cancer death in women, with mammographic screening the only modality shown to decrease the death rate. However, only 17% to 41% of women have ever been screened, and multiple barriers to screening have been identified. This study examined physician and patient factors at a single encounter to explore components influencing mammography ordering. METHODS: Ten family physicians in a primary care research network completed daily data cards on encounters with women presenting for annual examinations, chronic problems, or breast-related complaints. Information collected included patient age, personal or family history of breast cancer, physician's perception of expected compliance, previous mammogram results, breast examination, physician's perception of need for a mammogram, whether the mammogram was ordered, and the patient's method of payment for the test. RESULTS: Eight hundred thirty-nine patients were entered into the study, and 277 mammograms were ordered. Mammograms were ordered for a greater percentage of patients with insurance (36%) than for those without insurance (26%) (P < .001). A multivariate analysis indicated that several factors helped to correctly classify 90% of mammogram ordering: the patient was making a first visit, a breast-related visit, or a visit for an annual examination; the patient had had a previous mammogram; had a breast examination at the current visit or within the past year; and the physician believed the patient would comply and believed that a mammogram was indicated. CONCLUSIONS: Factors unique to a physician-patient visit influence the physician with regard to ordering a mammogram, including the type of visit, whether the physician believes a mammogram is indicated, and the cost. PMID- 8409890 TI - Effects of the Patient Self-Determination Act on patient knowledge and behavior. AB - BACKGROUND: In effect since 1991, the Patient Self-Determination Act (PSDA) requires that institutions receiving government insurance payments document that they have informed patients of their right to decide on life-preserving measures. Implementing the PSDA should make discussion of advance directives a routine part of acute care hospital admissions. Yet the proportion of those actually completing advance directives such as living wills remains relatively small. METHODS: A telephone survey questionnaire was administered to patients who were hospitalized before and after the implementation of the PSDA. Survey questions probed patient knowledge about living wills and behavior toward obtaining living wills. RESULTS: Patient knowledge about advance medical directives correlated positively with race (white), income (> or = $10K), and level of education (high school or more). Moreover, a significantly greater number of patients hospitalized after implementation of the PSDA knew about living wills than the number of those hospitalized before the Act's implementation. However, actually obtaining a living will correlated positively with age (> 36 years) alone, and implementation of the PSDA was not related to the number of patients who obtained a living will. CONCLUSIONS: Although the study results show that the measures the hospital in the study used to meet PSDA requirements increased patient awareness of living wills, they failed to increase the number of patients who act on this awareness. This finding indicates that simply informing patients about their right of self-determination is insufficient to meet the intended goals of the legislation. PMID- 8409891 TI - Prevalence and predictors of problem drinking among primary care diabetic patients. AB - BACKGROUND: Alcohol abuse among patients with diabetes mellitus is dangerous and complicates therapy, but its prevalence and the factors that predict it are unknown. This study examined the prevalence of problem drinking among a large number of primary care diabetic patients, exploring its relation to age, race, sex, psychological factors, and other health behaviors. METHODS: Volunteers with insulin-dependent diabetes mellitus and non-insulin-dependent diabetes mellitus were surveyed at three primary care practice sites. Patients completed a health risk appraisal designed to elicit alcohol use and other health practices, and two psychometric instruments: the Brief Encounter Psychosocial Instrument and the Affect Balance Scale. Fasting blood glucose and hemoglobin A1C levels were also determined. RESULTS: Of 395 diabetic patients, 32 (8.1%) had a drinking problem as defined by answering yes to the question "Have you ever had a drinking problem?" or reporting their last drink to be within 24 hours, or both. Patients with a drinking problem coped less well with psychological stress and had a more highly negative affect than those without a drinking problem. Depression, black race, and male sex were significantly associated with problem drinking (odds ratios = 8.42, 2.70, and 3.80, respectively). Problem drinking did not predict glycemic control but was associated with smoking and less frequent glucose monitoring. CONCLUSIONS: The prevalence of problem drinking among patients with diabetes mellitus appears lower than among other medical outpatient populations and is in keeping with the prevalence found in community surveys. While the lack of association between problem drinking and glycemic control in diabetic patients may be surprising, these data help define the characteristics of this subgroup of diabetic patients and highlight the need for family physicians to intensify alcohol screening efforts in this population. PMID- 8409892 TI - Perspectives on patient-doctor communication. AB - Until recently, the content, structure, and function of communication between doctors and patients has received little attention and has been excluded from the realm of scientific inquiry; as a result, most clinicians have had little formal training in communication skills. In this paper leaders in doctor-patient communication present four approaches that are currently used as the basis for clinical training and research, summarize the progress made in forming a consensus, and outline the implications of these perceptions for practicing physicians. PMID- 8409893 TI - Coagulopathy presenting as calf pain in a racquetball player. AB - Most cases of lower leg pain in athletes result from musculoskeletal injury. Occasionally these patients do not respond to treatment in a timely fashion. This should alert the clinician to rethink the original diagnosis and consider more unusual causes of leg pain. Deep venous thrombosis must be considered in a young athletic person experiencing unexplained persistent calf pain after exercise. Further investigation may be necessary to rule out a hereditary or acquired hypercoagulable state. PMID- 8409894 TI - Nutrition-related prevention: an interdisciplinary strategy. PMID- 8409895 TI - Shooting the dinosaur. PMID- 8409896 TI - Cold exposure and food restriction facilitate physiological responses to short photoperiod in Djungarian hamsters (Phodopus sungorus). AB - We investigated the influence of ambient temperature (Ta) and food availability on seasonal timing and extent of physiological responses to short photoperiod (SP), in particular daily torpor, in Djungarian hamsters (Phodopus sungorus). Exposure of hamsters to cold temperature (Ta = 5 degrees C), relative to warm Ta (23 degrees C), resulted in: 1) a significant advance (P < 0.05) of the first occurrence of torpor among cold-exposed hamsters (days 52-97 vs. days 83-99 in SP); 2) a higher (P < 0.01) incidence of torpor (48% vs. 20% torpid animals/day); 3) a higher (P < 0.05) degree of molt into the winter pelt; and 4) an accelerated reduction of body weights (P < 0.001). However, within SP/cold-Ta exposed groups, individual hamsters clearly showed different tendencies for torpor (torpor on 0 95% of days observed). Therefore, we evaluated the effects of small changes in Ta on torpor frequency and extension by subjecting the same SP-adapted individuals to varying temperatures. Lowering of Ta from 15 degrees C to 10 degrees C and 5 degrees C caused significant (P < 0.05) increases in the incidence of torpor (20%, 33%, and 40%, respectively) and lower minimal body temperatures during hypothermia (P < 0.05). When the same animals were subjected to 24-48 h lasting periods of food restriction (60% of the ad libitum intake), torpor frequency further increased 1.8- to 2.6-fold at all Tas. These results show that Ta and food availability are effective in modifying both seasonal timing and extent of photoperiodically controlled adaptations. This integration of multiple environmental cues, combined with a pronounced within-species variability of winter adjustments, indicates that Djungarian hamsters are capable of flexible physiological responses towards unpredictable climatic changes in the environment. PMID- 8409897 TI - Specific phosphorylation of membrane proteins of Mr 44,000 and Mr 32,000 by the autophosphorylated insulin receptor from the hepatopancreas of the shrimp Penaeus monodon (Crustacea: Decapoda). AB - The insulin receptor, purified from the hepatopancreas of the shrimp Penaeus monodon, is a hydrophobic heterodimer of subunits with relative masses (Mr) of 70,000 and 58,000, as estimated by FPLC on Superose 12 and SDS-PAGE. Only the subunit of Mr 70,000 was autophosphorylated after the addition of insulin. The autophosphorylation occurred specifically at Tyr residues, as demonstrated by the specific subsequent dephosphorylation by the phosphotyrosyl protein phosphatase from the hepatopancreas of the shrimp Penaeus monodon. Proteins of Mr 44,000 and Mr 32,000 on the plasma membrane from the hepatopancreas of the shrimp Panaeus monodon were phosphorylated by the autophosphorylated insulin receptor from the shrimp hepatopancreas, but not by that from the human placenta. The detergent, Triton X-100, caused noticeable enhancement of the autophosphorylation of both shrimp and human insulin receptors. PMID- 8409898 TI - Prostaglandin E2 and prostaglandin F2 alpha involvement in the corticosterone and cortisol release by the female frog, Rana esculenta, during ovulation. AB - Interrenal and ovarian tissues of Rana esculenta were incubated in vitro during the preovulatory, ovulatory and postovulatory phases to study the basal release of prostaglandin E2 (PGE2), prostaglandin F2 alpha (PGF2 alpha), corticosterone, and cortisol. The effects of exogenous PGE2 and PGF2 alpha on interrenal and ovarian corticosteroid release were also studied. In addition, the plasma values of these four hormones were assessed during the same phases. During in vitro interrenal incubations, PGE2, PGF2 alpha, corticosterone, and cortisol basal releases were higher in the postovulatory phase, PGE2 and PGF2 alpha treatment in vitro increased corticosteroids during the ovulatory phase. During in vitro ovarian incubations, PGE2 basal release was higher in the preovulatory phase and PGF2 alpha and corticosteroids in the ovulatory phase; PGE2 treatment in vitro decreased corticosteroids in the ovulatory phase, and PGF2 alpha increased corticosteroids in the preovulatory and postovulatory phases. PGE2, corticosterone and cortisol plasma values were higher during the postovulatory phase, while PGF2 alpha was elevated during the ovulatory phase. These findings suggest that ovarian corticosteroids could be considered one of the factors inducing ovulation and that their synthesis may be modified by PGs. PMID- 8409899 TI - The wound epithelium of regenerating limbs of Pleurodeles waltl and Notophthalmus viridescens: studies with mAbs WE3 and WE4, phalloidin, and DNase 1. AB - The wound epithelium of regenerating limbs of the American newt, Notophthalmus viridescens (Nv), up-regulates a number of antigens, including those recognized by mAbs WE3 and WE4. In the present study, we show that the WE3 antigen is up regulated in a similar fashion in the wound epithelium of the European newt, Pleurodeles waltl (Pw). mAb WE3 and WE4 reactivities to secretory/transport body cell types, including integumentary glands, perineurium, endothelium, and conjunctiva, are also similar in these two species of newt. However, mAb WE4 reacts to both the epidermis and wound epithelium in Pw, whereas in Nv, mAb WE4 reacts only to the wound epithelium. Because the WE3 antigen is cytoskeleton associated and Western blots reveal a 43 kDa species, we compared mAb WE3 reactivity with that of rhodamine-labeled phalloidin, a known actin-binding compound. Phalloidin did not react preferentially to the wound epithelium, conjunctiva, or other cell types strongly reactive to mAb WE3. Pretreatment of sections and tissue extracts with DNAse 1, a protein known to bind to actin, nearly abolished mAb WE3 reactivity in tissue sections and both WE3 and WE4 reactivity in ELISA assays, respectively. The results lead to the hypothesis that the WE3 and WE4 antigens are actin-binding proteins unique to the wound epithelium and other secretory/transport cell types. PMID- 8409900 TI - Pedicle and antler development following sectioning of the sensory nerves to the antlerogenic region of red deer (Cervus elaphus). AB - Sensory nerves supplying the deer antlerogenic region were sectioned about 60 days prior to pedicle initiation to determine the extent of neural influence on pedicle and first antler growth. Our results from a combination of histological examination and immunohistochemical localization showed that all 12 antlerogenic regions were successfully deprived of sensory nerve supply, but in 10 of 12 cases there was partial regeneration during the experimental period. In the two cases where no sensory reinnervation occurred, pedicle growth did not show any differences compared with partially sensory reinnervated or intact pedicles. With or without reduced sensory nerve supply, first antlers were initiated, grown, cleaned of velvet, cast, and regenerated in the normal way, but they were smaller than controls. Consequently, we conclude that a sensory nerve supply is not necessary for normal pedicle formation and for the first antler cycle, but plays a role in determining antler size. PMID- 8409901 TI - Studies of lysophospholipids related to the hamster sperm acrosome reaction in vitro. AB - Phospholipase A2 and lysophospholipids have been implicated in the mammalian sperm acrosome reaction. In this study we further investigated the role of this enzyme and lysophospholipids on the acrosome reaction of hamster spermatozoa. Hamster epididymal spermatozoa were incubated under capacitation and acrosome reaction-inducing conditions. After 3.0 and 3.5 h, the spermatozoa were treated with different doses of lysophosphatidylcholine for 12 min. Then the percentage of motility, hyperactivation, and acrosome reaction was evaluated by light microscopy. Lysophosphatidylcholine, 10 micrograms/ml, was the highest acrosome reaction-inducing dose without an effect on sperm motility. Lysophosphatidylcholine induced the acrosome reaction only when added to spermatozoa capacitated for a minimum of 2 h. This effect was apparent after 1 min of its addition and reached a plateau after 5 min. Lysophosphatidylethanolamine and lysophosphatidylinositol were also effective in inducing the acrosome reaction. Lysophosphatidylserine did not have any effect on the reaction, but caused an increase in sperm hyperactivation. Sperm treated with the phospholipase A2 inhibitors quinacrine dihydrochloride and p-bromophenacyl bromide showed an inhibition of the spontaneous occurrence of the acrosome reaction. These inhibitors, however, did not block the acrosome reaction induced by lysophosphatidylcholine. The time course of the lysophosphatidylcholine induced acrosome reaction was the same whether control or inhibitor treated spermatozoa were used. These results suggest that the membrane events of the acrosome reaction initiate with the activation of the phospholipase A2, thus producing the fusogen agents necessary for this exocytotic event. PMID- 8409902 TI - Characterization of equine oviductal proteins synthesized and released at estrus and at day 4 after ovulation in bred and nonbred mares. AB - Proteins synthesized and released in vitro by oviducts collected from horse mares during estrus and at day 4 after ovulation for bred and nonbred mares were examined by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS PAGE) and fluorography. Ampullary and isthmic regions both produced a wide array of nondialyzable proteins in culture. Major proteins or groups of proteins identified according to relative molecular weight (kDa) and apparent isoelectric point (pI) were at 100 kDa, pI 8; 100-200 kDa, pI 6; 150 kDa, pI 4.5; 60-100 kDa, pI 4; and an array of polypeptides at 21-22 kDa, pI 5-6. Oviductal secretory activity, measured by incorporation of radiolabeled amino acids into nondialyzable macromolecules released into incubation medium, was greater (P < .01) for the ampullary than the isthmic oviductal region. No consistent differences were observed in fluorograms between estrus vs. day 4 after ovulation, ampulla vs. isthmus, ipsilateral vs. contralateral to the corpus luteum or largest follicle, oviducts from bred vs. nonbred mares, or mare ages. Dialyzed medium from ampullary and isthmic regions of oviducts was subjected to 1 D or 2-D SDS PAGE followed by western blotting utilizing an antiserum directed against human retinol binding protein (RBP). The family of 21-22 kDA polypeptides was identified as immunoreactive RBP. PMID- 8409903 TI - Culture of intact Sertoli/germ cell units and isolated Sertoli cells from Squalus testis. II. Stimulatory effects of insulin and IGF-I on DNA synthesis in premeiotic stages. AB - To investigate growth control mechanisms during spermatogenesis in vitro, [3H]thymidine incorporation into acid-insoluble macromolecules was used to quantify DNA synthesis in cultured spermatocysts (intact Sertoli cell/germ cell clones) derived from premeiotic (PrM), meiotic (M), and postmeiotic (PoM) regions of dogfish (Squalus acanthias) testis. Forty-eight hours after seeding in basal medium, DNA synthesis was > 7-fold higher in PrM cysts than in other stages, thus verifying the staging procedure. In autoradiograms, germ cells of PrM cysts (e.g., spermatogonial and preleptotene stages) were labeled all-or-none, but not all cysts were labeled, and later developmental stages (e.g., cysts with round or elongating spermatids) were never labeled. Fetal bovine serum (FBS, 10%) and insulin-transferrin-selenite (ITS, 10 micrograms-10 ng/ml) doubled DNA synthesis in PrM cyst cultures but had no effect at other stages. Bovine insulin (10 micrograms/ml) and human recombinant insulin-like growth factor-I (IGF-I, 15 ng/ml) also doubled [3H]thymidine uptake in PrM cultures, but lower doses were less effective and estradiol-17 beta, transferrin, adult shark serum, purified shark relaxin, and a variety of other known growth factors were neither stimulatory nor inhibitory at the doses and conditions tested. Sertoli cell monolayers derived from PrM- or M-stage spermatocysts displayed a dose-response increase in DNA synthesis after addition of IGF-I (15-75 ng/ml), with a maximal increment significantly greater than with 10% FBS. Using [3H]thymidine incorporation by PrM cysts as an end-point, stimulatory bioactivity was detected in the < 30,000 kDa fraction of spent media from PrM Sertoli cells, whereas the low molecular weight fraction of M-stage Sertoli cells was inhibitory. Gel electrophoretic analysis of the two fractions revealed qualitative and quantitative differences in protein banding patterns, reinforcing the view that secretory activity of Sertoli cells is stage related. Results of these studies implicate insulin/IGF-I in mechanisms governing proliferation of male germ cells and support the view that Sertoli cells have an autocrine or paracrine role as both targets and sources of growth regulatory factors. PMID- 8409904 TI - Who is to serve? The chronically ill patient in the managed care milieu. PMID- 8409905 TI - Insurance coverage and usage of preventive health services. AB - The 1991 Florida Behavioral Risk Factor Survey which included 2,246 respondents to a random-digit-dial telephone survey, showed that 19.2% of Florida residents aged 18 and older reported they had no health insurance, and 24.7% reported they had no insurance covering outpatient services. Lack of insurance coverage was reported more frequently by younger adults (70% under age 40 vs 93% over age 60), by those with less than a high-school education (69% vs 85% for those with some college), by members of racial-ethnic minority groups (Hispanics 67.5%, Blacks 71.5%, whites 85%), and by residents of southeastern Florida. Persons without insurance coverage were less likely to report having had a check-up by a physician in the last year, more likely to report they needed to see a physician but could not because of cost (37% vs 9%), and less likely to have received a mammogram (33% vs 56%) or Pap smear (51.6% vs 67.8%) in the last year. These findings support the need for health-care reform to include assurance that health insurance covers preventive services. PMID- 8409906 TI - Vibrio vulnificus from raw oysters. Leading cause of reported deaths from foodborne illness in Florida. AB - Seventy-two cases of Vibrio vulnificus infection from raw oysters were reported from 1981-1992; 36 (50%) patients died, making this infection the leading cause of reported deaths from foodborne illness in Florida. The bacterium naturally occurs in coastal waters and may contaminate legally harvested and properly handled shellfish. Infection, usually by ingestion of contaminated raw oysters, can cause severe illness especially in individuals with preexisting liver disease. They are at 80 times greater risk of illness and over 200 times greater risk of death. The case fatality rate (63%) among patients with liver diseases was over 2.5 times the rate (23%) among those without liver disease. Infections usually occurred during the warm weather months and presented as fulminant septicemia, often complicated by necrotizing cutaneous lesions. Early treatment with antibiotics, debridement and amputation when necessary may improve survival. Prevention relies upon educating patients regarding risk and thorough cooking of shellfish. PMID- 8409907 TI - Biomedical waste rules. Their intent, scope, and difference. PMID- 8409908 TI - Myocardial preconditioning. Endogenous protection against infarction. AB - Even with heroic procedures, it is not yet possible to prevent cell death during a myocardial infarction; however, preconditioning has revealed an endogenous process capable of providing significant protection to the heart from such a fatal result. Mechanisms by which preconditioning operates have not yet been unraveled but its resolution has clear clinical significance. PMID- 8409909 TI - Winding up business. PMID- 8409910 TI - Hassle free zone. PMID- 8409911 TI - Detection of seven species of pathogenic leptospires by PCR using two sets of primers. AB - Two sets of primers derived from genomic DNA libraries of Leptospira serovars icterohaemorrhagiae (strain RGA) and bim (strain 1051) enabled the amplification by PCR of target DNA fragments from leptospiral reference strains belonging to all presently described pathogenic Leptospira species. The icterohaemorrhagiae derived primers (G1/G2) enabled amplification of DNA from L. interrogans, L. borgpetersenii, L. weilii, L. noguchii, L. santarosai and L. meyeri, whereas the bim-derived primers (B64-I/B64-II) enabled the amplification of L. kirschneri. Southern blot and DNA sequence analysis revealed inter-species DNA polymorphism within the region spanned by primers G1 and G2 between L. interrogans and various other Leptospira species. Using a mixture of primer sets G1/G2 and B64-I/B64-II, leptospires of serovars icterohaemorrhagiae, copenhageni, hardjo, pomona, grippotyphosa and bim were detected in serum samples collected from patients during the first 10 days after the onset of illness. PMID- 8409912 TI - Molecular analysis of a flagellar core protein gene of Serpulina (Treponema) hyodysenteriae. AB - The flaB2 gene encoding a protein located in the core of the periplasmic flagella of Serpulina hyodysenteriae was cloned and sequenced. The FlaB2 protein consists of 285 amino acids and has a calculated molecular mass of 31.1 kDa. Southern blot analysis indicated that at least one, and possibly two genes related to flaB2 are present in the genome of S. hyodysenteriae. Comparison of the amino acid sequence of FlaB2 to sequences present in data banks showed significant similarity with the core flagellins of other spirochaetes, in particular with a FlaB2 protein from Treponema phagedenis. PMID- 8409913 TI - The Escherichia coli serA-linked capsule locus and its flanking sequences are polymorphic, genetic evidence for the existence of more than two groups of capsule gene clusters. AB - Two families of Escherichia coli capsules, termed groups I and II, have been defined previously on the basis of a number of biochemical and genetic criteria. Recently, a third group of capsules, termed I/II has been suggested on the basis of chemical structure and mode of expression. In this paper, we show that group I capsule-producing strains lack the serA-linked group II capsule genes. In addition, group I/II capsule-producing strains lack the group II capsule genes despite the former genes also mapping near to serA. Therefore, the genetic data presented in this paper support the existence of three groups of capsule gene clusters, two of which are linked to serA. Sequences flanking the K4 capsule genes were found in the chromosome of all E. coli strains examined and were sometimes present in multiple copies at different loci, indicating that this chromosomal region is highly polymorphic. PMID- 8409914 TI - Cloning, nucleotide sequence and expression in Escherichia coli of a gene (ompM) encoding a 25 kDa major outer-membrane protein (MOMP) of legionella pneumophila. AB - A genomic library derived from a virulent isolate of Legionella pneumophila was constructed in Escherichia coli JM 83 using the cloning vector pUC19. The clones were screened by filter immunoassay using L. pneumophila rabbit polyclonal antisera and in the absence of in situ bacterial lysis one such clone, LP 116, expressed L. pneumophila-specific antigens on the surface of E. coli. Restriction endonuclease digest analysis and agarose gel electrophoresis revealed a fragment measuring approximately 750 bp. Southern hybridization confirmed that the fragment was L. pneumophila DNA. Sequencing data showed that the fragment was 810 bp in length with an open reading frame (ORF) of 678 bp. The outer-membrane profiles of the E. coli parent, the L. pneumophila DNA-contributing strain and clone LP 116 were compared by SDS-PAGE. A protein of 25 kDa was found in outer membrane preparations of both the clone LP 116 and L. pneumophila but not in E. coli JM 83. This was in agreement with the molecular mass of the deduced peptide of the mature protein. Immunoblots using L. pneumophila-specific polyclonal antiserum confirmed that this 25 kDa outer-membrane protein (OMP) was a L. pneumophila polypeptide. Both direct immunofluorescence assay and immunoblots using the commercially produced monoclonal antibody specific for the common antigen of the major outer-membrane protein (MOMP) confirmed that the 25 kDa protein produced by LP 116 was involved with the MOMP complex. The gene encoding this protein has been designated ompM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409915 TI - Actinobacillus pleuropneumoniae RTX-toxins: uniform designation of haemolysins, cytolysins, pleurotoxin and their genes. AB - The three different pore-forming RTX-toxins of Actinobacillus pleuropneumoniae are reviewed, and new and uniform designations for these toxins and their genes are proposed. The designation ApxI (for Actinobacillus pleuropneumoniae RTX-toxin I) is proposed for the RTX-toxin produced by the reference strains for serotypes 1, 5a, 5b, 9, 10 and 11, which was previously named haemolysin I (HlyI) or cytolysin I (ClyI). This protein is strongly haemolytic and shows strong cytotoxic activity towards pig alveolar macrophages and neutrophils; it has an apparent molecular mass in the range 105 to 110 kDa. The genes of the apxI operon will have the designations apxIC, apxIA, apxIB, and apxID for the activator, the structural gene and the two secretion genes respectively. The designation ApxII is proposed for the RTX-toxin which is produced by all serotype reference strains except serotype 10 and which was previously named App, HlyII, ClyII or Cyt. This protein is weakly haemolytic and moderately cytotoxic and has an apparent molecular mass between 103 and 105 kDa. The genes of the apxII operon will have the designations apxIIC for the activator gene and apxIIA for the structural toxin gene. In the apxII operon, no genes for secretion proteins have been found. Secretion of ApxII seems to occur via the products of the secretion genes apxIB and apxID of the apxI operon. The designation ApxIII is proposed for the nonhaemolytic RTX-toxin of the reference strains for serotypes 2, 3, 4, 6 and 8, which was previously named cytolysin III (ClyIII), pleurotoxin (Ptx), or macrophage toxin (Mat).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409916 TI - Migration of rat peritoneal cells after intra-abdominal infection with Bacteroides fragilis and Escherichia coli. AB - A fibrin clot model for intra-abdominal abscess formation was used to study the migratory properties of peritoneal cells from rats during the early stages of infection. Peritoneal cells and fibrin clot remnant were harvested 6 h after implantation of a sterile, singly infected (Escherichia coli or Bacteroides fragilis) or mixed infected (E. coli and B. fragilis) fibrin clot. Histological study of fibrin clots, removed 6 h after implantation, showed a deeper infiltration by host cells of B. fragilis infected clots compared to the others. This difference in infiltration by peritoneal cells was not due to differences in fibrinolytic activity of the bacterial strains. Differential cell counts of the peritoneal cells from rats implanted with sterile, singly and mixed infected fibrin clots showed distribution over subpopulations to be independent of the bacterial content of the infected clots used. In vitro migration assays showed no significant differences in migration by peritoneal cells from rats implanted with clots containing a different bacterial composition. Since B. fragilis infected fibrin clots were more deeply infiltrated by host defence cells than the other clots, and only mixed infected clots led to persistent abscesses in this model, we conclude that local conditions within the fibrin matrix rather than intrinsic cellular capacities of the host cells are important for the process of abscess formation. PMID- 8409917 TI - Interaction of Propionibacterium acnes with skin lipids in vitro. AB - Propionibacterium acnes is the predominant microbial resident within the pilosebaceous follicles of sebum-rich areas of human skin. This study investigated the effects of known hydrophobic components of sebum on the physiology and nutrition of this microorganism, grown anaerobically at 33 degrees C, under defined conditions using continuous culture techniques. The medium used was chemically defined, comprising eight amino acids, with glucose as the main carbon energy source, and the culture pH was maintained at 5.6. The range of sebum lipids assayed was based on the C18 monounsaturated fatty acid 9-cis octadecenoic acid (oleic acid). Stock micronized solutions were aseptically pulsed into continuous cultures in the presence and absence of glucose, and nutritional effects monitored. None of the lipid substrates significantly affected P. acnes growth either in terms of maximum specific growth rate (mu max) or final culture biomass yield. Glycerol (3 mg ml-1) was found to be a poor carbon/energy source in comparison to glucose. Bacterial cells did, however, adhere with varying degrees, to the different lipid species, with maximum adherence occurring with the free fatty acid. This observation was confirmed by preliminary uptake experiments using [14C]oleic acid. The interactive site for cell adherence may be the lipid-fibrillar layer associated with the cell surface of P. acnes, as discerned in electron microscopical studies. The findings of this investigation suggest that one function of the P. acnes lipase may be to aid colonization within the pilosebaceous follicle, by promoting cell adherence to components such as oleic acid. PMID- 8409918 TI - The DNA sequence and minimal replicon of the Corynebacterium glutamicum plasmid pSR1: evidence of a common ancestry with plasmids from C. diphtheriae. AB - The complete nucleotide sequence of pSR1, a 3 kb multicopy cryptic plasmid from Corynebacterium glutamicum ATCC 19223 has been determined. pSR1 is unrelated to the 4.4 kb Brevibacterium lactofermentum plasmid pBL1 and shows no DNA sequence conservation with plasmids from Staphylococcus. Transposon insertion and deletion mutants located the minimal replicon to within a 2.1 kb NcoI-BclI restriction fragment. This region contains a single large open reading frame, ORF2, flanked at the 5' end by a series of inverted repeat sequences which may modulate its expression, and at the 3' end by a region which may contain a replication origin. ORF2 (position 1633-2636) with a maximum coding potential of 36 kDa is essential for pSR1 replication and was designated the rep gene. The predicted ORF2 protein product exhibits 47% identity over a length of 343 amino acids with a replication associated ORF in the C. diphtheriae plasmid pNG2, many of the changes being in the third base position. This observation suggests that pSR1 and pNG2, which are two plasmids from environmentally separated Corynebacterium species, may share a common ancestral rep gene. PMID- 8409919 TI - The ancestral IncP replication system consisted of contiguous oriV and trfA segments as deduced from a comparison of the nucleotide sequences of diverse IncP plasmids. AB - In most plasmids which have been studied to date the functions required for plasmid replication are clustered in a 2-3 kb region. However, in all known naturally occurring plasmids of the Escherichia coli incompatibility group P the essential replication functions, oriV, the vegetative replication origin and trfA, which encodes proteins essential to activate oriV, are separated by blocks of DNA consisting of either known genes conferring resistance to antimicrobial agents and/or putative transposable elements. Nucleotide sequence comparisons reported here reveal that these blocks of DNA have inserted at different points into a backbone of DNA common to IncP plasmids. The results indicate that in the common ancestor of present IncP plasmids oriV and trfA must have been contiguous, whilst a rho-independent transcriptional terminator, now lost in IncP alpha plasmids, may have prevented trfA operon transcription from interfering with the activity of oriV. PMID- 8409920 TI - Characterization of an IS-like element from Mycobacterium tuberculosis. AB - A DNA sequence, present in members of the Mycobacterium tuberculosis complex, has been identified and characterized. The distribution of this DNA sequence among mycobacterial species was analysed by DNA hybridization and PCR experiments. As the sequence was detected only in bacteria belonging to the M. tuberculosis complex, it may be useful for the rapid discrimination of mycobacteria. Interestingly, the sequence has some characteristics of an insertion element (IS) and codes for a hypothetical protein with significant homologies to proteins encoded by several IS elements of other organisms, namely IS427 and IS869 from Agrobacterium tumefaciens, IS402 from Pseudomonas cepacia, Tn4811 from Streptomyces lividans and ISRm4 from Rhizobium meliloti. Together, these elements form a previously unrecognized family of transposable elements. This finding suggests the possibility of horizontal gene transfer between pathogenic mycobacteria and other organisms including Gram-negative plant-pathogenic bacteria. PMID- 8409921 TI - Cloning, nucleotide sequence and expression in Streptomyces lividans and Escherichia coli of pabB from Lactococcus lactis subsp. lactis NCDO 496. AB - A gene (pabB) encoding the aminase activity of p-aminobenzoate (PABA) synthase in Lactococcus lactis subsp. lactis was cloned in pIJ41 and expressed in Streptomyces lividans strains defective in PABA biosynthesis. Expression of the gene was associated with a 1.2 kb deletion between the aph promoter and the cloning site in pIJ41. Subcloning in pBR322 and expression in Escherichia coli AB3295 of the cloned L. lactis DNA fragment localized the pabB-complementing gene in a 1.9 kb segment. The nucleotide sequence of this segment contained a 1410 bp open reading frame encoding a 470-amino-acid polypeptide of 50937 Da. The deduced amino acid sequence showed substantial similarity to those reported for PabB and TrpE from several organisms. Synonymous codon usage reflected the low G + C content in the genomic DNA of L. lactis subsp. lactis, and therefore differed markedly from the preferred usage in the S. lividans host. The cloned heterologous pabB DNA was expressed in amounts that allowed accumulation of excreted PABA in cultures of S. lividans transformants. PMID- 8409922 TI - Mutants of Escherichia coli affected in respiration: the cloning and nucleotide sequence of ubiA, encoding the membrane-bound p hydroxybenzoate:octaprenyltransferase. AB - A mutant of Escherichia coli has been isolated that is unable to grow aerobically on non-fermentable substrates, but able to grow anaerobically on glycerol with alternative electron acceptors such as fumarate. Nitrate as electron acceptor supports anaerobic growth on glycerol, but not on succinate or lactate. Oxygen consumption rates by cell-free extracts with succinate, lactate or glycerol 3 phosphate as substrates were low relative to activities in an isogenic control strain but were restored in vitro by adding ubiquinone-1. Transformation of the mutant with a cloned 2.6 kb ClaI-PvuII fragment of chromosomal DNA restored cellular quinone levels and growth on succinate. The plasmid also complemented a previously isolated ubiA mutant for aerobic growth on non-fermentable substrates. The nucleotide sequence of the cloned fragment revealed a fragment of plsB (91.7 min on the E. coli chromosome map) and three open reading frames (ORFs), one of which (ORF3) encodes a protein with a predicted molecular mass of 32511 Da. The hydrophobicity profile of the ORF3 protein is characteristic of a membrane protein with five hydrophobic regions and is very similar to that of the Saccharomyces cerevisiae COQ2 gene product (p hydroxybenzoate:polyprenyltransferase, required for the second step of ubiquinone biosynthesis) and to the product of the E. coli cyoE gene. Complementation of ubi mutants with various deletion derivatives of the cloned DNA fragment confirms that ORF3 is ubiA. ORF3 is closely linked to ubiC (ORF2), which encodes chorismate lyase. PMID- 8409923 TI - Purification and properties of cyanide hydratase from Fusarium lateritium and analysis of the corresponding chy1 gene. AB - The filamentous fungus Fusarium lateritium is cyanide tolerant, due, at least in part, to the induction by cyanide of the enzyme formamide hydrolyase (EC 4.2.1.66). This enzyme, more commonly known as cyanide hydratase, catalyses the hydration of cyanide to formamide. The enzyme was purified from F. lateritium and showed a subunit molecular mass of 43 kDa (as judged by SDS-PAGE), while the native protein appeared to form aggregates of up to 1217 kDa (as judged by gel filtration and non-denaturing PAGE). mRNA samples from cultures grown with and without cyanide were in vitro translated and immunoprecipitated. This demonstrated that, in this species, the gene encoding the enzyme designated chy1, is cyanide inducible. Differential screening was used to isolate a cyanide hydratase cDNA clone which was subsequently used to obtain the corresponding genomic clone. A fragment of the cDNA clone encoding all but the first seven amino acids of the protein was expressed in E. coli using the expression vector pGEX-2T. Features of F. lateritium cyanide hydratase together with an analysis of the nucleotide sequence encoding this enzyme are presented. PMID- 8409924 TI - Identification of the formate dehydrogenases and genetic determinants of formate dependent nitrite reduction by Escherichia coli K12. AB - The formate dehydrogenases of Escherichia coli involved in electron transfer from formate to nitrite (Nrf activity: nitrite reduction by formate) have been identified. No previously undescribed selenoprotein was detected in bacteria grown under conditions optimal for the expression of Nrf activity. The Nrf activities of single mutants defective in either FdhN or FdhH were between 50 and 60% that of the parental strain. A double mutant defective in both FdhN and FdhH retained less than 10% of the activity of the FdhN+ FdhH+ strain. No Nrf activity was detected in a triple mutant defective in FdhN, FdhH and FdhO or in the selC strain. It is concluded that all three of the known formate dehydrogenases of E. coli can contribute to the transfer of electrons from formate to the Nrf pathway. Mutants defective in Nrf activity and cytochrome c552 synthesis were isolated by insertion mutagenesis or identified amongst strains received from the E. coli Genetic Stock Center. The mutations were located in at least three regions of the chromosome, including the 92 to 94 minute region which includes fdhF, the gene encoding FdhH required for formate hydrogenlyase activity. Fine structure mapping by P1 transduction established that the nrf mutations in the fdhF region were due to defects in three separable loci, all of which were independent of but close to fdhF. Clones were isolated from a cosmid library that complemented a deletion extending from fdhF into a region essential for Nrf activity. From these clones, plasmids were isolated that complemented only some of the Nrf- mutations in the 92 to 94 minute region, confirming the presence of different operons essential for Nrf activity and cytochrome c552 synthesis in this region. Suggested reasons for this genetic complexity include the need for proteins involved in electron transfer from the various formate dehydrogenases to cytochrome c552, for the attachment of the haem group to the apocytochrome and for cytochrome c552 export into the periplasm. PMID- 8409925 TI - Membrane-associated NADH dehydrogenase activities in Rhodobacter capsulatus: purification of a dihydrolipoyl dehydrogenase. AB - The presence of several NADH dehydrogenase activities associated with cytoplasmic membrane vesicles of chemoheterotrophically grown Rhodobacter capsulatus MT1131 was demonstrated by combining isoelectric focusing with NADH tetranitrobluetetrazolium activity staining, a procedure that should have general applicability in the analysis of bacterial NADH dehydrogenase activities. Low pI (pI = 5.7), Mid pI (pI = 6.9) and High pI (pI = 8.5) bands were resolved. The Mid pI NADH dehydrogenase activity was purified and identified as a dihydrolipoyl dehydrogenase. Our data indicate that this dihydrolipoyl dehydrogenase is derived from a 2-oxoacid dehydrogenase complex which is associated with the cytoplasmic membrane. PMID- 8409926 TI - Molybdenum uptake in Escherichia coli K12. AB - Molybdenum uptake was examined in Escherichia coli K12 using the radionuclide 99Mo. The molybdenum uptake system was characterized in an unusual chlD strain, which appeared to be normal in uptake of the MoO4(2-) ion but altered in subsequent molybdenum processing. As a consequence, molybdenum could be chased from cells in the chlD strain, while it was irreversibly assimilated in the wild type strain. Molybdenum uptake showed a biphasic kinetic curve, with a very rapid binding followed by a slow uptake phase. The uptake appeared to involve an active transport system. Molybdenum, probably in the form of molybdate, accumulated by a factor of about 30 in the cells. An energy source was necessary and uptake was inhibited by arsenate, but not by CCCP (carbonyl cyanide m chlorophenylhydrazone). The uptake system saturated with a Km of 2.5-2.7 x 10(-8) M. Uptake seemed to depend on a periplasmic binding protein, since cold shock treatment and arsenate abolished uptake. A molybdate binding protein activity was detected in the periplasmic fluid with a KD of 9 nM. Sulphate inhibited uptake and the uptake activity was pH dependent, with an apparent pK of 6.7. These results imply that molybdate transport belongs to the family of energy-dependent periplasmic binding protein systems. An explanation for the peculiar behaviour of the chlD strain used in this work is proposed. PMID- 8409927 TI - Cell wall assembly in Staphylococcus aureus: proposed absence of secondary crosslinking reactions. AB - The distribution of muropeptides formed by muramidase digestion of peptidoglycan from Staphylococcus aureus H was determined by gel-filtration HPLC. The observed crosslinking pattern supports the conclusion that incorporation of peptidoglycan in S. aureus proceeds by a similar mechanism to that proposed earlier for Bacillus megaterium. In this mechanism single glycan-peptide strands are incorporated into the sacculus by crosslinking reactions that take place only between the monomer muropeptide units of the incoming glycopeptide and muropeptides present in the innermost region of wall at the wall-membrane interface: such crosslinking reactions take place only during incorporation and no other crosslinking reactions occur. This assembly process has now been termed restricted monomer addition. The present analysis shows that the distribution of muropeptides in S. aureus peptidoglycan is in excellent agreement with that predicted by this mechanism. We propose that cell wall assembly in S. aureus proceeds via restricted monomer addition without any requirement for the secondary crosslinking reactions that have been suggested to occur in this organism. The high degree of crosslinking in S. aureus, 80% in this study, may result mainly from the freedom for crosslinking provided by the pentaglycine bridge peptide. PMID- 8409928 TI - Biochemical analysis of germination mutants to characterize germinant receptors of Bacillus subtilis 1604 spores. AB - Spores of Bacillus subtilis 1604 can be induced to germinate by incubation in L Ala (the ALA pathway) or in a combination of beta-D-glucose (Glc), beta-fructose (Fru), L-Asn and K+ (the GFAK pathway). Biochemical analysis of the germination response of a gerA mutant deficient in the ALA pathway revealed that L-Ala can replace L-Asn in the GFAK pathway (the GFAlaK pathway). In contrast to the ALA pathway of both the wild-type and of a gerB mutant, the GFAlaK pathway was insensitive to D-Ala and showed the same overall inhibitor profile as the GFAK pathway of wild-type and gerA spores. It is deduced that a second L-Ala receptor with different characteristics to that functioning in the ALA pathway is present in wild-type spores. Analysis of the germination response of a gerB mutant showed that whilst the rate of ALA germination could be stimulated by Glc as well as by Fru in the presence of Glc, the spores could not germinate in GFAK. In addition, Glc and Fru were unable to reverse D-Ala inhibition of L-Ala germination which they do in the wild-type. Thus, in the gerB mutant, the L-Ala/L-Asn receptor in the GFAK pathway is defective. It is concluded that the germination receptors in the ALA and GFAK pathways can functionally interact with each other to initiate B. subtilis spore germination. This conclusion is discussed in relation to proposed models of triggering of spore germination. PMID- 8409929 TI - The role of amniotic passage in the egg-adaptation of human influenza virus is revealed by haemagglutinin sequence analyses. AB - Obtaining an isolate of a human influenza virus in the allantoic cavity of the embryonated hen's egg is more efficient if the clinical sample is initially passaged in the amniotic cavity. To investigate the extent to which the variants present after allantoic propagation are also selected by amniotic passage, clinical virus passaged once in the amnion has been subjected to extensive genetic and antigenic analyses. The data indicate that the natural virus can replicate unrestrictedly within the amnion. However, exposure of amniotic virus to the allantois during the incubation period, which will occur through the hole between the amniotic and allantoic cavities caused by the inoculating needle, allows for the possibility of an egg-adapted variant establishing replication within the allantois and returning to the amnion. These observations illustrate why prior passage in the amnion increases the probability of a variant successfully establishing itself during a subsequent allantoic passage. PMID- 8409930 TI - Mouse hepatitis virus spike and nucleocapsid proteins expressed by adenovirus vectors protect mice against a lethal infection. AB - Infection with the mouse hepatitis coronavirus (MHV) provides an excellent model for the study of viral diseases of the central nervous system and the gastrointestinal tract. With the ultimate aim of studying mucosal immunity to MHV we have cloned the genes encoding the structural proteins of MHV strain A59 (MHV A59) into the E3 region of a human adenovirus type 5 vector. Infection of HeLa cells with the resulting recombinant adenoviruses AdMHVS, AdMHVN and AdMHVM revealed the correct expression of the spike (S), nucleocapsid (N) and membrane (M) proteins, respectively. Intraperitoneal inoculation of BALB/c mice with the recombinant viruses elicited serum antibodies which specifically recognized the respective MHV proteins in an immunoprecipitation assay. Only antibodies to the S protein neutralized MHV-A59 in vitro but titres were low. When analysed by ELISA or by immunofluorescence only the antibody response to the N protein was significant; weak responses or no detectable response at all were found for S and M, respectively. Upon intracerebral challenge with a lethal dose of MHV-A59 we found that a significant fraction of animals vaccinated with adenovirus vectors expressing either the S protein or N protein were protected. This protective effect was significantly stronger when the animals were given a booster immunization with the same vector prior to challenge. No protection was induced by AdMHVM. Interestingly, enhanced protection resulted when AdMHVS and AdMHVN were applied in combination as compared to survival after single immunizations. The results indicate that both the N and S proteins generate a protective immune response and suggest that this response is enhanced by combined expression of the two proteins. PMID- 8409931 TI - Coxsackievirus B1-induced murine myositis: no evidence for viral persistence. AB - The persistence of coxsackievirus B1 in the muscles of mice with coxsackievirus B1-induced chronic myositis was investigated. Neonatal CD1 Swiss mice were inoculated with a myositis-causing variant of coxsackievirus B1 (Tucson strain). Hamstring muscle samples of diseased mice obtained at various times after inoculation were examined for the presence of infectious virus, viral RNA and histological abnormalities. Viral RNA was detected up to 4 weeks after initiation of infection, whereas virus could be isolated from hamstring muscles for up to 2 weeks. Thereafter no sign of infection was demonstrated although histological abnormalities remained present for the entire observation period of 16 weeks. That viral RNA was detectable for only 2 weeks after tissues became negative for infectious virus suggests that the infection slowly waned rather than the viral RNA persisting. Hence, it is concluded that coxsackievirus B1 plays an essential role in the initiation of myositis but not in the maintenance of the chronic phase. PMID- 8409932 TI - Detection of unintegrated human immunodeficiency virus type 1 DNA in persistently infected CD8+ cells. AB - The presence of unintegrated viral DNA has been reported in cells persistently infected by lentiviruses, including human immunodeficiency virus type 1 (HIV-1). We confirm that CD8+ cells can be productively and persistently infected by HIV-1 for up to 4 months, as determined by secretion of viral core antigen p24 into the extracellular medium and by indirect immunofluorescence. The expression of the external viral glycoprotein gp120 at the surface of these cells was demonstrated by two-colour flow cytometry. Progeny virions recovered from CD8+ cells were infectious in CD4+ T cells. Despite an absence of significant cytopathology, these chronically infected CD8+ cells were shown to harbour unintegrated HIV-1 DNA, as detected by quantitative PCR. Both linear and circular forms of the extrachromosomal viral genome were present in infected CD8+ cells, as early as 3 weeks before a peak in viral replication. These findings provide evidence that the presence of unintegrated viral DNA during lentiviral infection may not always correlate with c.p.e. PMID- 8409933 TI - Analysis of endoproteolytic cleavage and intracellular transport of human immunodeficiency virus type 1 envelope glycoproteins using mutant CD4 molecules bearing the transmembrane endoplasmic reticulum retention signal. AB - We investigated endoproteolytic processing of the human immunodeficiency virus (HIV) envelope glycoprotein precursor, gp160, as well as envelope-mediated membrane fusion in the presence of CD4 molecules that were either partially or fully retained in the endoplasmic reticulum (ER). Pulse-chase analyses revealed that gp160 formed complexes with CD4 molecules, and gp160 in the complex was endoproteolytically cleaved to gp120 and gp41 in the secretory pathway. The gp120/gp41 complex thus generated was properly targeted to the plasma membrane in cells expressing gp160 and wild-type CD4 or mutant CD4 molecules that were partially retained in the ER. Additionally, membrane fusion (syncytium) assays were performed to monitor the presence or absence of gp120/gp41 complexes at the cell surface of cotransfected cells and demonstrated that the HIV-1 envelope glycoprotein-mediated membrane fusion was appreciably reduced in the presence of wild-type CD4 or either one of the mutant CD4 molecules. Reduction in the formation of syncytia appears to be due predominantly to saturation of the CD4 binding site on the gp120/gp41 complex at the cell surface of cotransfected cells, but partial retention of the complex in the ER could also partly account for the reduction. However, the intracellular gp120/gp41 complex generated in cells expressing gp160 and CD4 mutant having the transmembrane ER retention signal (KKTC) was completely retained in the ER and hence could not participate in membrane fusion events at the plasma membrane. Taken together, these data suggest that the endoproteolytic cleavage of gp160 occurs in the ER or cis-Golgi network, and ER retention strategies can potentially be used in preventing the spread of HIV-1 infection in permissive cells. PMID- 8409934 TI - A recombinant retrovirus carrying a non-producer human immunodeficiency virus (HIV) type 1 variant induces resistance to superinfecting HIV. AB - A human immunodeficiency virus (HIV) type 1-infected Hut-78 cell clone (F12) shows a peculiar phenotype: it exhibits an altered viral protein pattern, is a nonproducer and is resistant to homologous superinfection. To determine whether this phenotype is dependent upon the expression of the HIV-1 genome integrated therein, the SstI/SstI F12 provirus [deprived of HIV long terminal repeats (LTRs)] was cloned and inserted in the pLj retroviral vector bearing the neomycin (neo) and Geneticin resistance gene. CD4+ HIV-susceptible CEMss cells (a CEM clone able to form large syncytia 2 to 3 days post-HIV infection) were infected with the recombinant retroviruses rescued from the F12/HIV-pLj-transfected (in either sense or antisense orientation) amphotropic packaging cells PA 317. Neo sense resistant gene clones showed approximately 10 copies of viral DNA/cell (without detectable major deletions) only in episomal form, low viral RNA expression and a viral protein pattern characterized by an uncleaved gp160, no gp41 and little, if any, p55 gag precursor (as in F12 cells). Superinfection of these F12/HIV DNA-engineered clones with HIV-1 resulted in a significant reduction in the yield of superinfecting HIV. This effect (more pronounced when the clones were maintained under neo selective pressure) was observed in all five retrovirus-infected clones exhibiting the presence and expression of sense episomal F12/HIV DNA but not in two clones bearing an antisense F12/HIV DNA or in one clone bearing only the pLj vector. These results indicate that bio-engineered human CD4+ cells expressing the F12/HIV genome exhibit a significant resistance to HIV superinfection. PMID- 8409935 TI - CD8+ cytotoxic T lymphocytes against antigenic variants of caprine arthritis encephalitis virus. AB - Cytotoxic T lymphocytes (CTL) specific for caprine arthritis-encephalitis virus (CAEV) were characterized using a colorimetric immunocytochemistry assay to measure surviving target cells. Peripheral blood lymphocytes from a CAEV-infected goat were cytotoxic to autologous, CAEV-infected dermal fibroblast target cells following in vitro stimulation of the lymphocytes with CAEV antigen. The lymphocytes were not cytotoxic to infected allogeneic target cells or to mock infected autologous or allogeneic target cells. This CAEV antigen-specific, major histocompatibility complex-restricted cytotoxicity was mediated by CD8+ lymphocytes as demonstrated by selective depletion with anti-CD8 antibody and complement. CTL primed with one isolate of CAEV (CAEV-Co) had no detectable activity against target cells infected with either of two neutralization variants and diminished activity against target cells infected with three other neutralization variants. This apparent variability of CTL-sensitive epitopes among CAEV isolates may contribute to CAEV escaping immune control and may complicate vaccine design. PMID- 8409936 TI - Kinetics of infectivity are dissociated from PrP accumulation in salivary glands of Creutzfeldt-Jakob disease agent-inoculated mice. AB - The protease-resistant isoform of prion protein (PrP) has been implicated in the pathogenesis and transmission of Creutzfeldt-Jakob disease (CJD), scrapie and other related diseases, but the relationship between the infectious agent and PrP awaits elucidation. In the present study, we have examined levels of infectivity together with accumulation of the protease-resistant form of PrP (PrPCJD) in various tissues of CJD agent-inoculated mice. Accumulation of PrPCJD occurred only in tissues, including brain, salivary gland and spleen, in which infectivity was readily detectable throughout the course of the experiment. The brain showed the highest levels of both infectivity and PrPCJD accumulation, with well correlated kinetics. On the other hand, the high titres of infectivity detected in salivary gland and spleen early after inoculation of the agent were obviously distinguishable from PrPCJD. Furthermore, in the salivary gland, the kinetics of infectivity and the accumulation of PrPCJD reversed; infectivity declined as PrPCJD accumulated in the tissue. Our findings indicate that PrPCJD accumulation is associated with replication of the agent; however, PrPCJD is unlikely to be the agent itself. PMID- 8409937 TI - Duplicated genes within the variable right end of the genome of a pathogenic isolate of African swine fever virus. AB - The right variable region of the genome of a pathogenic strain of African swine fever virus (ASFV), Malawi LIL20/1, has been sequenced and 15 open reading frames (ORFs) identified by computer analysis. Eight of these ORFs were found to be similar to previously described ASFV ORFs and three of these belong to two previously described multiple gene families (MGF), 360 and 110. Four of the remaining five ORFs belong to a novel MGF, designated MGF 100, and the last ORF encodes a protein that is similar to the virus structural protein, p22. Copies of MGF 110 and the gene coding for p22 have previously been characterized only at the left end of the ASFV genome. The organization of these genes suggests evolution by duplications, deletions and sequence transposition from one end of the genome to the other. Sequence comparisons of members of MGF 360 suggest that the Malawi LIL20/1 genome has undergone separate DNA rearrangements compared to the Ba71V genome. Lastly, one ORF was found to be similar to the myeloid differentiation primary response protein, MyD116 and to the herpes simplex virus neurovirulence-associated factor ICP34.5. PMID- 8409938 TI - Multiple pathways for gene activation in rodent cells by the smaller adenovirus 5 E1A protein and their relevance to growth and transformation. AB - By immunoprecipitating protein products from virus-infected baby rat kidney (BRK) cells with specific antibodies, we found that the smaller, 243 residue (243R) E1A protein of human adenovirus 5 (Ad5) activated expression of the virus genes for E1B 55K, E2A 72K, E3 19K, hexon, fibre and penton base and the cellular gene for PCNA. The 243R protein also activated the E2A 72K gene in several rodent cell lines. In transient expression assays, this protein trans-activated the E2 early and major late promoters, suggesting that its effect was at least partially transcriptional. Similar assays with mutants of the E2 early promoter suggested that the ATF- and distal E2F-binding sites were required for this activation. Using mutant viruses with deletions in E1A, we found evidence for three separate pathways by which the 243R protein activated gene expression: one depended on sequences in exon 1 required for this protein to bind to p300, a second depended on sequences in exon 1 required for the protein to bind to pRb and the third appeared to be independent of exon 1 altogether and to depend on exon 2. The relative importance of these pathways for activation varied with the gene and cell. We conclude that a major role of E1A in the transformation of BRK cells by Ad5 is to activate specific genes by at least the first two pathways. PMID- 8409939 TI - Identification of the gene encoding the major capsid protein of fish lymphocystis disease virus. AB - The gene encoding the major capsid protein (MCP) of fish lymphocystis disease virus (flounder isolate; FLCDV-f) has been identified by PCR using oligonucleotide primers corresponding to different regions of the MCP of Tipula iridescent virus (TIV), iridescent virus 22 (IV22) and Chilo iridescent virus (CIV). DNA fragments of 0.4 kbp, 0.5 kbp and 0.27 kbp in size were amplified using oligonucleotide primers corresponding to amino acids (aa) 146 to 153 (primer 1) and 274 to 268 (primer 6), or aa 146 to 153 (primer 1) and 313 to 304 (primer 8), or aa 304 to 312 (primer 7) and 385 to 381 (primer 9) of the MCP of TIV, respectively. The PCR products were used as hybridization probes for screening the gene library of FLCDV-f. The MCP gene of FLCDV-f(1377 bp; 459 aa; 51.4K) was identified within the DNA sequence of the EcoRI FLCDV-f DNA fragment C (11.2 kbp; 0.611 to 0.718 map units). A high degree of aa sequence identity/similarity was detected between the MCP of FLCDV-f and TIV (50.3%/33.8%), IV22 (49.1%/34.2%). CIV (53%/29.5%) and African swine fever virus (16%/38.1%). PMID- 8409940 TI - The complete sequence and gene organization of the short unique region of herpesvirus of turkeys. AB - The DNA sequence of the whole of the short unique region (U(s)) and that of part of the short terminal repeat (TRs) of herpesvirus of turkeys (HVT) were determined. HVT U(s) is 8.6 kbp long and contains eight potential open reading frames (ORFs). Seven of these have counterparts in the U(s) of herpes simplex virus type 1 (HSV-1). The homologous proteins include US1, US2, US10, protein kinase (US3) and the glycoproteins gD, gI and gE. In addition, HVT contains one ORF which has a counterpart in the U(s) of Marek's disease virus (MDV) but is not homologous to any other known herpesvirus gene. Although HVT and MDV proteins encoded by U(s) genes have evident similarities with proteins encoded by alphaherpesviruses, multiple alignment analysis of predicted amino acid sequences show that HVT proteins are more closely related to MDV proteins than to homologous proteins of mammalian alphaherpesviruses. The percentage amino acid identity between HVT and MDV U(s)-encoded proteins ranges from 35 to 65, the most conserved protein being encoded by the homologues of the HSV-1 US2 gene. Most of the genes are colinear with those of HSV-1 except US10 which is transposed in HVT and MDV. A characteristic feature of HVT is the fact that approximately two thirds of the gE gene is located in the inverted repeats flanking U(s). PMID- 8409941 TI - The herpes simplex virus type 1 DNA polymerase accessory protein, UL42, contains a functional protease-resistant domain. AB - Herpes simplex virus type 1 encodes its own DNA polymerase (Pol), the product of the UL30 gene, and a polymerase accessory subunit, the product of the UL42 gene, both of which are required for viral DNA replication. Pol and the UL42 protein associate to form a heterodimeric complex (Pol/UL42) which is more active and has a higher processivity than the Pol catalytic subunit alone. The Pol/UL42 complex has been reconstituted by mixing together highly purified Pol and UL42 subunits obtained from recombinant baculovirus-infected cells. We have used polymerase activity on poly(dA):oligo(dT20), a template that the Pol subunit utilizes with low efficiency, to measure the formation of the Pol/UL42 complex. Our data indicate that the association constant for the Pol/UL42 complex is 1 x 10(8) M-1. Proteolytic digestions of UL42 were performed to determine whether structural domains of UL42 could be disclosed by differential amino acid accessibilities. The ability of these protease-resistant domains to form a functional complex with Pol was determined by measuring their ability to stimulate Pol activity on poly(dA):oligo(dT20). We have found that trypsin digestion of UL42 in the presence of DNA generates protease-resistant fragments of 28K and 8K which co elute from a MonoQ column and are able to stimulate Pol activity on poly(dA):oligo(dT20). Complex formation of the 28K and 8K tryptic fragments with Pol was also shown by their co-immunoprecipitation with antibody to Pol. It was determined that the 28K fragment of UL42 comprised amino acids 1 to 245 or 1 to 254 of UL42, whereas the 8K fragment started at amino acid 255. Thus, controlled proteolysis of UL42 revealed two closely contiguous structural domains that retained the ability to complex with Pol and stimulate Pol activity. PMID- 8409942 TI - Autocrine secretion of interferon-alpha/beta and tumour necrosis factor-alpha synergistically activates mouse macrophages after infection with herpes simplex virus type 2. AB - Resistance of mice to infection with herpes simplex virus type 2 (HSV-2) is strongly dependent on the function of macrophages (M phi). Infection of mouse M phi with HSV-2 results in an early (4 to 10 h) activation of the cells with an enhanced respiratory burst generated after membrane triggering with a phorbol ester. The role of monokines produced during this infection was analysed. Both interferon-alpha/beta (IFN-alpha/beta) and tumour necrosis factor-alpha (TNF alpha) were produced within the very first hours after infection of M phi with HSV-2. Exogenously added IFN-alpha/beta conferred to M phi a respiratory burst capacity comparable to that seen after virus infection, whereas TNF-alpha by itself was unable to prime M phi for a respiratory burst. In fact concentrations of TNF-alpha comparable to those found in HSV-2-infected M phi cultures generally suppressed the response. However, when TNF-alpha was added together with IFN alpha/beta a dose-dependent synergistic enhancement of the IFN-induced M phi activation was seen. The kinetics of the synergistic activation by the two monokines was similar to that seen with IFN-alpha/beta alone. Neutralizing antibodies to IFN-alpha/beta and TNF-alpha were able to diminish the HSV-induced priming of M phi for a respiratory burst. When the two antibodies were used together in subneutralizing concentrations an additional diminution of the responsiveness was seen, indicating that both monokines are involved in the virus induced priming of M phi. However, high concentrations of antibodies to IFN alpha/beta alone were able to abolish the activation completely, whereas this was not the case with anti-TNF-alpha. Collectively these data demonstrate that autocrine secretion of IFN-alpha/beta by M phi infected with HSV-2 is a sine qua non for the activation of M phi during the infection, and that this effect of IFN is synergistically enhanced, also in an autocrine manner, by TNF-alpha. It is suggested that this reciprocal M phi-monokine interaction may be of importance in resistance to virus infections. PMID- 8409943 TI - Deleting two amino acids in glycoprotein gI of pseudorabies virus decreases virulence and neurotropism for pigs, but does not affect immunogenicity. AB - The virulence, pathogenicity and immunogenicity of two pseudorabies virus (PRV) variants were investigated in 3-week-old pigs that had been intranasally infected. Variant M303 (delta 125,126) lacked amino acids valine (125) and cysteine(126) in an immunodominant antigenic region of glycoprotein I (gI) containing two discontinuous antigenic domains, whereas M304 (delta 59,60) lacked amino acids glycine(59) and aspartic acid(60) in a continuous antigenic domain. M303 (delta 125,126) was not virulent for pigs, but M304 (delta 59,60) was as virulent as wild-type PRV: all pigs died within 8 days of infection. Both gI mutant viruses replicated in the oropharyngeal mucosa, although M304 (delta 59,60) replicated to higher virus titres than M303 (delta 125,126), and virus was recovered from various tissues. However, in contrast to M304 (delta 59,60), M303 (delta 125,126) was not recovered from any central nervous system (CNS) tissues examined. Thus, the tendency of PRV to locate in the CNS was markedly reduced by deleting amino acids valine(125) and cysteine(126) of gI. Pigs immunized with M303 were completely protected against challenge infection; no clinical signs of disease were detected, no virus was shed, and no secondary antibody response was detected. Thus, deleting amino acids valine(125) and cysteine(126) in gI decreases virulence and neurotropism and does not affect immunogenicity. PMID- 8409944 TI - Complete nucleotide sequence of pepper huasteco virus: analysis and comparison with bipartite geminiviruses. AB - The complete nucleotide sequence of pepper huasteco geminivirus (PHV) isolated in Northern Mexico was determined. The PHV genome consists of two circular ssDNA molecules of 2631 bases (PHV A) and 2589 bases (PHV B). PHV has a genome organization typical of a bipartite geminivirus with four open reading frames (ORFs) (AR1, AL1, AL2 and AL3) in component A and two (BR1 and BL1) in component B. An unexpected ORF was found in the complementary sense strand of PHV A. This ORF, termed AL5, is found entirely inside, but in the opposite orientation to AR1 (encoding the coat protein). AL5 shows some homology to equivalent but smaller ORFs predicted in other geminiviruses. Phylogeny trees based on pairwise comparisons of AR1, AL2, AL3, BL1 and BR1 predicted proteins placed PHV among the western hemisphere geminiviruses. A phylogeny tree based on AL1 (replicase encoding ORF), on the other hand, placed PHV with eastern hemisphere geminiviruses, i.e. African cassava mosaic virus and the Sardinia and Israel isolates of tomato yellow leaf curl virus. Possible mechanisms for the 'hybrid or transition nature' of PHV are discussed. PMID- 8409945 TI - Protoplasts transiently expressing the 200K coding sequence of cowpea mosaic virus B-RNA support replication of M-RNA. AB - In order to identify the viral polymerase involved in cowpea mosaic virus (CPMV) RNA replication the 87K, 110K and 170K proteins as well as the complete 200K polyprotein of CPMV B-RNA have been produced in cowpea protoplasts, using expression vectors based on the 35S promoter of cauliflower mosaic virus. CPMV specific proteins were obtained that were indistinguishable from proteins found in CPMV-infected protoplasts. Proteolytic processing of precursor proteins synthesized from the expression vectors proved that the 24K protease contained within these proteins is active. Moreover, it was established that protoplasts transfected with the expression vector containing the entire 200K coding sequence, but not those transfected with vectors containing the 170K, 110K or 87K coding sequences, were able to support replication of co-inoculated M-RNA. Despite the ability to support replication of M-RNA for protoplasts transiently expressing the 200K coding region, CPMV-specific RNA polymerase activity dependent on exogenous added template RNA could not be detected in extracts of these protoplasts in assays using poly(A).oligo(U) or other template/primer combinations. In contrast, extracts of protoplasts in which poliovirus polymerase was produced exhibited RNA polymerase activity in such assays. These results indicate that the CPMV polymerase, unlike the poliovirus polymerase, is not able to use oligo(U) as a primer or cannot function on exogenous template and primer RNA. PMID- 8409946 TI - Sequence data to settle the taxonomic position of bean common mosaic virus and blackeye cowpea mosaic virus isolates. AB - The nucleotide sequences of the coat protein genes and 3' non-translated regions (3'-NTRs) of three isolates of bean common mosaic virus (NL1, NL3 and NY15) and one isolate of blackeye cowpea mosaic virus (W) were determined. Comparison of these sequences revealed that the coat proteins of NL1, NY15 and W were identical in size (287 amino acids) and exhibited an overall sequence similarity (94 to 97%), and 84 to 98% in their N-terminal regions. Furthermore, their 3'-NTRs were very similar in length [253 to 256 nucleotides (nt)] and sequence (93 to 96% similarity). In contrast, the coat protein of NL3 had only 261 amino acids and showed 87 to 89% similarity with NL1, NY15 and W whereas its N-terminal region revealed only 46 to 61% similarity. The 3'-NTR of NL3 also displayed appreciable differences, both in length (240 nt) and sequence (56 to 63% similarity). These results, in combination with earlier serological findings, justify the conclusion that NL1, NY15 and W should be considered strains of the same virus, i.e. bean common mosaic virus, and that NL3 is a strain of a different potyvirus for which the name 'bean black root virus' is proposed. PMID- 8409947 TI - RNA sequence of potato virus X strain HB. AB - The genomic RNA of the potato virus X (PVX) strain HB, isolated in Bolivia and able to overcome all known resistance genes, has been cloned and sequenced. The PVXHB RNA sequence is 6432 nucleotides long and contains, similarly to the RNAs of other PVX strains, five open reading frames encoding proteins of M(r)s 165.1K, 24.5K, 12.4K, 7.6K and 25.1K (coat protein), respectively. Multiple amino acid sequence alignments of the coat proteins of four PVX strains identified eight amino acid residues unique for PVXHB. Structural prediction comparisons of the coat proteins of PVXHB and of the other strains suggest a general structural similarity. However, two of the eight amino acid residues unique for strain HB gave rise to a change in the predicted coat protein structure, suggesting a possible involvement in the resistance-breaking activity of PVXHB. PMID- 8409948 TI - Monoclonal antibodies to glycoprotein B differentiate human herpesvirus 6 into two clusters, variants A and B. AB - The distribution of glycoprotein B (gB) among different human herpesvirus 6 (HHV 6) strains was analysed with a panel of three monoclonal antibodies (MAbs) derived from mice immunized with U1102-infected lymphocytes. MAb 2D9 reacted specifically by immunofluorescence and immunoprecipitation with the U1102 and GS isolates, and failed to react with Z29 and the variant B strains Hashimoto and SF. In addition, Z29, Hashimoto and SF gB had a lower M(r) than U1102 and GS gB. MAb 2D9 also failed to react with the exanthem subitum isolate CV, included in this study as an as yet poorly characterized isolate. Consistent with this result, CV failed to react with the variant A-specific MAb to gp82-105 and behaved as a variant B virus even with respect to the diagnostic HindIII endonuclease restriction cleavage site located in a fragment hybridizing to the pZVH14 probe. By contrast with MAb 2D9, MAbs 2B9 and 2D10 reacted with all of the isolates tested, strengthening the argument tha they have common epitopes. Based on the antigenic and M(r) specificities of gB, the HHV-6 isolates tested were arranged into two non-overlapping clusters, which closely parallel the variant A and B strain groups, defined previously by several criteria, including restriction endonuclease polymorphism, antigenic variations, growth in in vitro cultures and sequence analyses. PMID- 8409949 TI - Topoisomerase I and II activities are required for Epstein-Barr virus replication. AB - The roles of topoisomerases I and II in Epstein-Barr virus (EBV) replication were investigated using Raji cells infected with EBV. The topoisomerase II inhibitor ellipticine inhibited the synthesis of EBV polypeptides at concentrations which did not affect total protein synthesis. Slot blot analysis of total cellular DNA showed that camptothecin and ellipticine inhibited replication of progeny EBV DNA in superinfected Raji cells at concentrations which did not inhibit synthesis of EBV early polypeptides prerequisite for EBV DNA replication. Analysis of the structure of EBV DNA termini demonstrated that both inhibitors affected the replicating EBV DNA. Gardella gel electrophoresis showed that both inhibitors affected the formation of the linear form of EBV DNA. However, restriction analysis of EBV DNA in superinfected Raji cells demonstrated that both inhibitors degraded neither endogenous nor exogenous EBV DNA. Cell viability was not affected by either inhibitor at the concentrations tested. These findings suggest that topoisomerase II is required for expression of the EBV genome and that both topoisomerases I and II are involved in replication of the EBV genome during the lytic phase of the life cycle. The effects of topoisomerase inhibitors on the circular form of EBV DNA during virus replication are discussed. PMID- 8409950 TI - Herpes simplex virus type 1 capsid protein, VP21, originates within the UL26 open reading frame. AB - The goal of experiments reported here was to identify the genes that encode capsid proteins VP21 and VP24 of herpes simplex virus type 1 (HSV-1). Capsids were isolated from infected cells and the proteins were separated by SDS-PAGE. N terminal amino acid sequence analysis of partial CNBr digestion products, and of intact VP21, showed that it is encoded within the UL26 open reading frame (ORF) of HSV-1 beginning with codon 248 and probably extending to the end of the ORF (codon 635). Similar analysis of digestion products confirmed that VP24 is specified by codons 1 to 247 at the 5' end of the UL26 ORF. Each of the seven known capsid proteins has now been assigned to an ORF. PMID- 8409951 TI - Interaction between bovine papillomavirus type 4 and cocarcinogens in the production of malignant tumours. AB - Bovine fetal palatine tissue infected with bovine papillomavirus type 4 (BPV-4) was implanted subcutaneously in athymic nude mice. The implants developed into cysts containing papillomas essentially the same as those in the natural host. In order to investigate the interaction of cocarcinogens with BPV-4 in cell transformation, the virus-infected implants were exposed in vivo to either the tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) or the tumour initiator 7,12-dimethylbenz[a]-anthracene (DMBA). Papillomas were detected in a greater number of infected implants in the presence of either TPA or DMBA than in the absence of either of these chemicals indicating interaction between the virus and these two agents. Moreover, malignantly transformed cells arose with high frequency from infected implants that had been exposed to either chemical. In the presence of chemical and absence of virus or vice versa no neoplastic changes were seen histologically, indicating that cooperation between virus and cocarcinogen is required for transformation. PMID- 8409952 TI - Molecular epidemiology of foot-and-mouth disease virus type O. AB - A phylogenetic tree based on the VPI sequences of type O foot-and-mouth disease virus (FMDV) has been derived. Direct sequencing of PCR products has been used to obtain the VP1 gene sequences of new isolates. The tree exhibits four main lineages that largely correlate with the geographical origin of isolates. The analysis supports a close relationship between European O1 field isolates and vaccine strains, with the exception of O Thalheim Aus/81 and O Wuppertal Ger/82 which were probably of non-European origin. Analysis of nucleotide substitutions indicates that synonymous mutations play a major role in FMDV evolution. PMID- 8409953 TI - Rhinoviral receptor discrimination: mutational changes in the canyon regions of human rhinovirus types 2 and 14 indicate a different site of interaction. AB - Amino acid sequence comparisons between the capsid proteins of several human rhinovirus (HRV) serotypes identified residues potentially involved in the discrimination between the major and the minor group receptors. Amino acids conserved within minor group HRVs were substituted in a full-length cDNA clone of HRV2 for those found at equivalent positions in major group HRVs. Transfection of HeLa cells with RNAs transcribed from seven individual mutated cDNAs gave rise to only two viable viruses; growth characteristics and affinity for the minor group receptor of both were unchanged compared to wild-type. Similar mutations in HRV14 were previously shown to alter the affinity for its receptor; the contact sites between the minor group viruses and the respective receptor may therefore be different. PMID- 8409954 TI - Non-viral sequences at the 5' ends of Dugbe nairovirus S mRNAs. AB - Analyses of the mRNA transcription processes of viruses in four genera (Bunyavirus, Hantavirus, Phlebovirus and Tospovirus) of the family Bunyaviridae have revealed a common mechanism of initiation using host-derived primers, known as cap-snatching. To provide similar information on the fifth genus in the family, the 5' ends of Dugbe nairovirus S mRNA species were specifically cloned and sequenced. This revealed the presence of non-viral heterogeneous sequences, five to 16 nucleotides in length (average of 10 nucleotides) at the 5' ends, confirming that cap-snatching to prime mRNA synthesis is a familial characteristic of the Bunyaviridae. Inspection of the sequences in the primers on nairovirus, bunyavirus and phlebovirus mRNAs suggests that in some cases polymerase slippage occurs shortly after initiation, resulting in a partial reiteration of the 5'-terminal nucleotides of the viral RNA. PMID- 8409955 TI - Isolation of a retrovirus from two fish cell lines developed from chinook salmon (Oncorhynchus tshawytscha) with plasmacytoid leukaemia. AB - Two new cell lines developed from chinook salmon with plasmacytoid leukaemia have been found to be producing a virus. The virus has been identified as a retrovirus based on: type of c.p.e. induced in culture; morphology and density of the particle; presence of Mn(2+)-dependent, poly(rA)-directed reverse transcriptase activity which was associated with a density of 1.16 to 1.18 g/ml in sucrose; electrophoretic pattern of the polypeptides from purified virions; elevated [3H]UTP labelling of RNA in the cell cultures occurring at a density of 1.16 to 1.18 g/ml in sucrose. This report describes the first isolation of a retrovirus from a salmonid cell line. PMID- 8409956 TI - Nucleotide sequence of beet cryptic virus 3 dsRNA2 which encodes a putative RNA dependent RNA polymerase. PMID- 8409957 TI - The future of psychiatry and the medical model. PMID- 8409958 TI - Has multiple personality disorder remained consistent over time? A comparison of past and recent cases. AB - The purpose of this study was to determine whether recent descriptions of multiple personality disorder are consistent with descriptions from the past. Clinical presentations and childhood histories obtained from early case reports of multiple personality disorder published between 1800 and 1965 (N = 52) were compared with recent case reports published in the 1980s (N = 54). Recent and past cases did not differ in age at diagnosis, length of treatment, duration of follow-up, presence of child and opposite gender personalities, and exposure to hypnosis. Recent cases differed significantly from past cases in mean number of personalities (12 vs. 3), age of onset (11 vs. 20 years), proportion of males (24% vs. 44%), and in prevalence of childhood abuse histories (81% vs. 29%). The authors discuss clinical and cultural factors that may have contributed to the change over time in the number of reported cases, complexity of personality structure, and description of etiological childhood trauma. Although a core set of symptoms has consistently been associated with this disorder over time, other important aspects have not been stable. PMID- 8409959 TI - Ethnic differences in psychiatric diagnosis among Asian American adolescents. AB - This is the first investigation of the psychiatric diagnosis of Asian American adolescents using data from the Los Angeles County Department of Mental Health. It was hypothesized that Asian American adolescents receive different diagnoses than Caucasian adolescents, and furthermore, that there are intra-Asian differences in diagnosis among the Asian subgroups. Asian American adolescents were categorized in the following subgroups: Chinese, Japanese, Korean, Filipino, Vietnamese, and other Asians (i.e., Southeast Asians and Pacific Islanders). Separate comparisons were made for male and female adolescents. The findings strongly support the presence of ethnic differences between Asian and Caucasian adolescents and also among Asian subgroups in both male and female groups. In the Asian-Caucasian comparison, Asian males and females received significantly more nonpsychiatric diagnosis than Caucasians. Asian males were more often diagnosed with nonpsychiatric disorder and less often with affective disorders than Caucasian males. Asian females were more frequently diagnosed with major depression and nonpsychiatric disorder than Caucasian females. Among Asian American adolescents, Chinese and Japanese adolescents received similar diagnoses while Korean and Vietnamese also showed similar patterns in diagnosis. The implications of intra-Asian differences are discussed. PMID- 8409960 TI - The prevalence and nature of lithium noncompliance among Chinese psychiatric patients in Hong Kong. AB - Based on the multiple criteria of serum lithium level, patients' subjective report and doctors' impression in the previous year, a noncompliance rate of 30% was found among 50 Chinese patients on chronic lithium treatment in Hong Kong. Of numerous variables analyzed, only lithium monotherapy, younger age, and earlier age of onset were significantly associated with noncompliance. Exploratory interviews documented the multidimensionality of lithium noncompliance. They argued against statistical attempts to establish a universal clinical formula for the prediction of lithium noncompliance, and underscored the personal, social and medical contexts in which illnesses were lived and treatments used. For cultural reasons, familiar Western rationales for legitimating lithium noncompliance, such as missing of highs, loss of assertiveness, loss of creativity, and excessive fear of weight gain, were rarely encountered. After 1 1/2-years of attendance and with specific interventions at the lithium clinic, the noncompliance rate decreased from 30% to 14%. The important role of a parent-child-dominated Chinese kinship system in the chronic care of psychiatric patients is also discussed. PMID- 8409961 TI - Using drugs to facilitate sexual behavior is associated with sexual variety among injection drug users. AB - Little normative data exist about the frequency and variety of sexual behavior of injection drug users. Sexual behaviors of a group of 313 injection drug users (225 men and 88 women) were assessed by a structured interview at the start of an acquired immune deficiency syndrome prevention project. Celibacy was reported by 12.3% of men and 26.9% of women in the year before the initial interview. Male injection drug users who had been sexually active reported a mean number of 4.61 (median, 2.0) female sexual partners in the previous year; sexually active women reported a mean number of 5.28 (median, 1.0) male sexual partners. Sexually active men and women reported median condom use frequency at 0% of vaginal intercourse events (mean for men, 10.31%; mean for women, 14.48%). Male injection drug users who reported using drugs to help them relax for sex, to enhance sexual performance, or to meet sexual partners reported greater frequency of anal intercourse, fellatio, and cunnilingus, less relative frequency of vaginal intercourse, more sexual partners, and greater involvement in being paid for sex and paying for sex than did men who did not report using drugs to enhance sex. Men who used drugs to enhance sex also reported higher frequency of use of a number of different drug classes than men who did not use drugs to enhance sex. For women, using drugs to enhance sex was related to greater frequency of anal intercourse and having more sexual partners. Implications for treatment of drug abusers and future research are discussed. PMID- 8409962 TI - The effects of social support on Hopkins Symptom Checklist-assessed depression and distress in a cohort of human immunodeficiency virus-positive and -negative gay men. A longitudinal study at six time points. AB - Questionnaire data were collected from a panel of 342 gay men at risk for acquired immune deficiency syndrome enrolled in the Coping and Change Study between 1985 and 1987, and 1988 and 1990. Data were obtained across a period of 5 years in six serial wave pairs to determine the relationship of social support to Hopkins Symptom Checklist-assessed subsequent depression and general distress and to investigate whether the trends observed were stable or transient over time. Both objective and subjective components of social support demonstrated dramatic within-person stability over time (r = .47 to .86). A measure of subjective social support was modestly but significantly associated with lower depression at four of the six time periods and a lower level of general distress at one time period. Before appropriately controlling for current depression, subjective social support appeared to account for up to a third of the variance in future depression; after such controls were included in the regression equation, it became apparent that the independent contribution of support only ranged from 4% to 6% across the study period. This emphasizes the importance of including current mental health in longitudinal analyses. The respondents' social participation and involvement with others did not affect either depression or general distress at any time during the study period. These results indicate that while social participation may have no effect, subjective social support appears to influence often mental health in this cohort. Furthermore, human immunodeficiency virus seropositive men may at times benefit from such support. PMID- 8409963 TI - Visual hallucinations and mental state. A study of 14 Charles Bonnet syndrome hallucinators. AB - Complex visual hallucinations are usually a sign of acute psychopathology or gross cognitive impairment, but may also occur in people with visual deficits- the Charles Bonnet Syndrome. The mental state of 14 Charles Bonnet hallucinators was assessed using four psychological tests: the Beck Depression Inventory, the State-Trait Anxiety Inventory, the Mini-Mult, and the Mini-Mental State Examination. Results are consistent with earlier reports suggesting that these hallucinations are not due to psychopathology or compromised cognitive functioning. It is proposed that these complex visual hallucinations represent ongoing neural activity in the visual system following eye damage. PMID- 8409964 TI - Recreational therapy and behavior management on inpatient units. Is recreational therapy therapeutic? PMID- 8409965 TI - Cultural shaping of illness: a longitudinal perspective on apparent depression. PMID- 8409966 TI - Differential ultrastructure of synaptic terminals on ventral longitudinal abdominal muscles in Drosophila larvae. AB - The innervation of ventral longitudinal abdominal muscles (muscles 6, 7, 12, and 13) of third-instar Drosophila larvae was investigated with Nomarski, confocal, and electron microscopy to define the ultrastructural features of synapse-bearing terminals. As shown by previous workers, muscles 6 and 7 receive in most abdominal segments "Type I" endings, which are restricted in distribution and possess relatively prominent periodic terminal enlargements ("boutons"); whereas muscles 12 and 13 have in addition "Type II" terminals, which are more widely distributed and have smaller "boutons". Serial sectioning of the Type I innervation of muscles 6 and 7 showed that two axons with distinctive endings contribute to it. One axon (termed Axon 1) has somewhat larger boutons, containing numerous synapses and presynaptic dense bodies (putative active zones for transmitter release). This axon also has more numerous intraterminal mitochondria, and a profuse subsynaptic reticulum around or under the synaptic boutons. The second axon (Axon 2) provides somewhat smaller boutons, with fewer synapses and dense bodies per bouton, fewer intraterminal mitochondria, and less developed subsynaptic reticulum. Both axons contain clear synaptic vesicles, with occasional large dense vesicles. Approximately 800 synapses are provided by Axon 1 to muscles 6 and 7, and approximately 250 synapses are provided by Axon 2. In muscles 12 and 13, endings with predominantly clear synaptic vesicles, generally similar to the Type I endings of muscles 6 and 7, were found, along with another type of ending containing predominantly dense-cored vesicles, with small clusters of clear synaptic vesicles. This second type of ending was found most frequently in muscle 12, and probably corresponds to a subset of the "Type II" endings seen in the light microscope. Type I endings are thought to generate the 'fast' and 'slow' junctional potentials seen in electrophysiological recordings, whereas the physiological actions of Type II endings are presently not known. PMID- 8409967 TI - Ultrastructure of neuromuscular junctions in Drosophila: comparison of wild type and mutants with increased excitability. AB - The ventral longitudinal muscles of the Drosophila larval body wall are innervated by at least four types of synaptic terminals that can be distinguished on morphological grounds at the light microscopical level. The innervation of these muscles has been previously shown to be regulated by neuronal activity. In this report we investigate the ultrastructural basis for synaptic bouton differences by using serial sections, and examine the structure of synaptic terminals in mutants with increased excitability. We report that individual identifiable muscle fibers are innervated by terminals containing two to three types of synaptic boutons that can be distinguished in terms of synaptic vesicle population, presynaptic and postsynaptic specialization, and general shape. We propose a model to account for the bouton types observed at the light microscopical level. We find that in the hyperexcitable mutant eag Sh, there are dramatic ultrastructural alterations at synaptic boutons. These alterations include a partial depletion of two types of synaptic vesicles and a change in appearance of a third type, changes in number and appearance of synaptic densities, and the presence of multivesicular bodies. Our results show that an increase in neuronal excitability produces profound effects in synaptic terminal structure. PMID- 8409968 TI - Biological studies of a putative avian muscle-derived neurotrophic factor that prevents naturally occurring motoneuron death in vivo. AB - A series of in vivo studies have been carried out using the chick embryo to address several critical questions concerning the biological, and to a lesser extent, the biochemical characteristics of a putative avian muscle-derived trophic agent that promotes motoneuron survival in vivo. A partially purified fraction of muscle extract was shown to be heat and trypsin sensitive and rescued motoneurons from naturally occurring cell death in a dose-dependent fashion. Muscle extract had no effect on mitotic activity in the spinal cord and did not alter cell number when administered either before or after the normal cell death period. The survival promoting activity in the muscle extract appears to be developmentally regulated. Treatment with muscle extract during the cell death period did not permanently rescue motoneurons. The motoneuron survival-promoting activity found in skeletal muscle was not present in extracts from a variety of other tissues, including liver, kidney, lung, heart, and smooth muscle. Survival activity was also found in extracts from fetal mouse, rat, and human skeletal muscle. Conditioned medium derived from avian myotube cultures also prevented motoneuron death when administered in vivo to chick embryos. Treatment of embryos in ovo with muscle extract had no effect on several properties of developing muscles. With the exception of cranial motoneurons, treatment with muscle extract did not promote the survival of several other populations of neurons in the central and peripheral nervous system that also exhibit naturally occurring cell death. Initial biochemical characterization suggests that the activity in skeletal muscle is an acidic protein between 10 and 30 kD. Examination of a number of previously characterized growth and trophic agents in our in vivo assay have identified several molecules that promote motoneuron survival to one degree or another. These include S100 beta, brain-derived neurotrophic factor (BDNF), neurotrophin 4/5 (NT-4/5), ciliary neurotrophic factor (CNTF), transforming growth factor beta (TGF beta), platelet-derived growth factor-AB (PDGF-AB), leukemia inhibitory factor (CDF/LIF), and insulin-like growth factors I and II (IGF). By contrast, the following agents were ineffective: nerve growth factor (NGF), neurotrophin-3 (NT3), epidermal growth factor (EGF), acidic and basic fibroblast growth factors (aFGF, bFGF), and the heparin-binding growth-associated molecule (HB-GAM).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8409969 TI - Accommodation of mouse DRG growth cones to electrically induced collapse: kinetic analysis of calcium transients and set-point theory. AB - Electrical stimulation causes growth cones of mouse dorsal root ganglion neurons to collapse. During chronic stimulation, however, growth cones resume motility. In addition, these growth cones are now resistant to the collapsing effects of subsequent stimulation, a process we term accommodation. We compared the kinetics of electrically induced Ca2+ transients in naive and accommodated growth cones in order to determine whether the accommodation process results from a change in the Ca2+ transient, or a change in the Ca2+ sensitivity of the growth cones. Three kinetics were determined: (1) the initial increase to peak Ca2+ levels produced by 10 Hz stimulation; (2) recovery from peak Ca2+ levels during stimulus trains lasting 15 min; and (3) clearing of Ca2+ from growth cones after terminating the stimulus. These kinetics were analyzed using single exponential fits to changes in fura-2 fluorescence ratios. The electrically evoked increase in Ca2+ was significantly slower in accommodated growth cones (tau = 6.0 s) compared to naive growth cones (tau = 1.4 s). Despite the slower increase of [Ca2+]i in accommodated growth cones, peak [Ca2+]i was similar to that reached in naive growth cones, and the steady-state Ca2+ level was significantly elevated after chronic stimulation. Thus, accommodated growth cones maintained outgrowth at [Ca2+]i that caused collapse initially. Time course experiments show that accommodation is a slow process (t 1/2 = about 3 h). Accommodation did not induce measurable changes in the rates of Ca2+ homeostasis during or after stimulus trains. The kinetics of Ca2+ recovery during (tau = 90 s) and after 15 min of stimulation (tau = 8.5 s) was not significantly different in accommodated versus naive growth cones. Rates of 45Ca2+ efflux were also similar in both types of growth cones. These results suggest two regulatory processes contributing to growth cone motility during chronic stimulation: (1) recovery of [Ca2+]i to levels permissive to neurite outgrowth, and (2) an increase in the range of optimal [Ca2+]i for growth cone motility. These adaptive responses of mammalian growth cones to chronic stimulation could be involved in the modulation of CNS development by electrical activity of neurons. PMID- 8409970 TI - Cues intrinsic to the retina induce nAChR gene expression during development. AB - Recent studies of optic nerve regeneration in goldfish have indicated that the optic tectum plays an important role in modulating the induction of nicotinic acetylcholine receptor (nAChR) gene expression in regenerating retinal ganglion cells (Heiber, Agranoff, and Goldman, 1992, J. Neurochem. 58:1009-1015). These observations suggest that induction of these genes is regulated by brain target regions. The appearance of nAChR mRNA in the developing rat retina coincides with a time when ganglion cells are sending axons to their brain targets (Hoover and Goldman, 1992, Exp. Eye Res. 54:561-571). Might a mechanism similar to that seen during goldfish optic nerve regeneration also mediate induction of nAChR gene expression during development of the mammalian retina? This possibility was tested by either transplanting embryonic rat retina to different brain regions, or explanting it to organ culture and assaying for nAChR gene expression. These studies showed that induction of the nAChR genes in developing rat retina is independent of the environment in which the retina develops. These results indicate that either the retinal microenvironment or a signal intrinsic to the retinal ganglion cell is responsible for this induction. PMID- 8409971 TI - Regeneration from crayfish phasic and tonic motor axons in vitro. AB - An explant culture system is described that allows examination of axonal growth from the tonically and phasically active motoneurons of the abdominal nerve cord of the crayfish. In this preparation, growth occurs from the cut end of the axon while the remainder of the motoneuron is undisturbed. In vitro growth from the branches of the third roots, which contain the axons from the tonic and phasic motoneurons of abdominal ganglia one through four, was verified as axonal by retrograde labeling of axons and neuronal somata within the nerve cord. Growth from the axons of phasic and tonic cells was observed as early as 24 h after plating and continued for an additional 7-10 days. The morphology and growth rates of the motor terminals differed between the tonic and phasic axons. The phasic axons grew significantly faster and branched more often than did the tonic motor axons. These differences in growth may be related to differences in motoneuron size or, may result from differences in electrical activity. Tonic motoneurons show spontaneous impulse activity for up to 6 days in culture, whereas phasic motoneurons show no spontaneous impulse activity. In addition, the differences in growth may be related to the morphological differences in tonic and phasic motor terminals observed in situ. PMID- 8409972 TI - Axial level-dependent differences in size of avian dorsal root ganglia are present from gangliogenesis. AB - Dorsal root ganglia (DRG) form from neural crest cells that colonize the rostral sclerotomies of the somites of the trunk of higher vertebrates. Differences in sensory field size and innervation density are reflected by the size of the ganglia at different axial levels. Some of the position-dependent differences in ganglionic size derives from asymmetry in embryonic programmed cell death, when more DRG neurons die at non-limb-innervating levels than at the levels that innervate the limbs. I have now examined early chick embryos to determine whether there is asymmetry in DRG size at the time of their condensation at stage 20 [embryonic day (E) 3], before the onset of cell death. Ganglia in brachial segments 14 and 15 are more than 80% larger on average than those in cervical segments 5 and 6 at this stage of development. This difference in volume is due to increased numbers of cells in the brachial sensory ganglia. Several other morphometric parameters of the DRG and sclerotomies were then determined. The rostro-caudal length was found to be significantly greater for brachial ganglia. The greater length of the brachial ganglia was found to be correlated with (1) a greater length of brachial than cervical sclerotomies and (2) the occupation by brachial ganglia of a larger proportion of the rostro-caudal extent of the sclerotome. These results demonstrate that the mature pattern of axial differences in ganglionic size are foreshadowed by a pattern set during the period of gangliogenesis, which is then further sculpted by apoptosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8409973 TI - Synaptic repression at crayfish neuromuscular junctions. II. Evidence for calcium involvement. AB - The inability of synaptic junctions to generate normalisized postsynaptic potentials under normal physiological conditions was studied at crayfish neuromuscular synapses. Synaptic repression in the superficial flexor muscle system of the crayfish was induced by surgery: the nerve was cut in the middle of the target field, and the lateral muscle fibers were removed. After this surgery, the remaining medial synapses were unable to generate normal-sized junction potentials (jp) over the medial muscle population. In an attempt to study the mechanisms underlying this response, we varied the extracellular calcium concentration of the Ringers solution bathing the preparation, in both repressed and control animals, while monitoring the size of the same junction potential. The junction potential generated by the spontaneous activity of the nerve increased in size with increasing calcium concentrations in control animals, but failed to do so in repressed animals, that is, changes in external calcium concentrations did not affect repressed synapses. However, in the presence of the calcium ionophore A23187, control and repressed synapses both show an increase in the junction potential sizes they generate. Our data suggest that calcium is involved in the mechanisms that underlie synaptic repression in this crustacean neuromuscular system. PMID- 8409974 TI - Effects of development and altered gravity conditions on cytochrome oxidase activity in a vestibular nucleus of the larval teleost brain: a quantitative electronmicroscopical study. AB - The mitochondrial enzyme, cytochrome oxidase, was localized cytochemically in the nucleus magnocellularis, a primary relay nucleus of vestibular information within the area octavolateralis in the fish brain. Larvae of the cichlid fish Oreochromis mossambicus were analyzed at different developmental stages (4, 10, and 35 days post-hatching) and after long-term exposure (8 days) to increased gravity (2-4 g). Quantification of highly reactive, moderately reactive, and nonreactive mitochondria reveals differences in the cytochrome oxidase activity of various cellular structures, for example, perikarya of neurons, presynaptic terminals, and myelinated and nonmyelinated cell profiles. Cytochrome oxidase activity in the mitochondria of neuronal perikarya increases during development which parallels the differentiation of the area octavolateralis. This possibly reflects the increasing energy demand during maturation and innervation of the magnocellular nucleus. Hyper-g-exposure of the larvae for 8 days (centrifuge) caused a further augmentation of cytochrome oxidase activity in the perikarya within the nucleus magnocellularis. This may reflect an increased oxidative metabolism resulting from the need for compensation of altered inputs from gravity-sensitive epithelia in the inner ear. Another possibility is that acceleration within a centrifuge causes physiological stress for the animals and, therefore, influences the cytochrome oxidase activity in neurons. PMID- 8409975 TI - Interactions between spinal cord stimulation and activity blockade in the regulation of synaptogenesis and motoneuron survival in the chick embryo. AB - The present study investigated the effects of spinal cord stimulation, neuromuscular blockade, or a combination of the two on neuromuscular development both during and after the period of naturally occurring motoneuron death in the chick embryo. Electrical stimulation of the spinal cord was without effect on motoneuron survival, synaptogenesis, or muscle properties. By contrast, activity blockade rescued motoneurons from cell death and altered synaptogenesis. A combination of spinal cord stimulation and activity blockade resulted in a marked increase in motoneuron death, and also altered synaptogenesis similar to that seen with activity blockade alone. Perturbation of normal nerve-muscle interactions by activity blockade may increase the vulnerability of developing motoneurons to excessive excitatory afferent input (spinal cord stimulation) resulting in excitotoxic-induced cell death. PMID- 8409976 TI - Effects of the steroid hormone 20-hydroxyecdysone and prior sensory input on the survival and growth of moth central olfactory neurons in vitro. AB - Neurons in the developing (antennal) olfactory lobe of the moth Manduca sexta undergo a period of extensive process outgrowth and branching that coincides temporally with both a rising titer of the steroid hormone 20-hydroxyecdysone and the ingrowth of sensory axons from receptors in the antenna. To evaluate the contribution of these two influences to the morphological development of antennal lobe neurons, we placed the neurons in cell culture. Antennal-lobe neurons were dissociated from normal and chronically unafferented lobes at different stages of development and were exposed to different doses of hormone. Six neuronal cell types with distinctive and stable morphologies appeared in cultures from all stages of pupal development. Morphological changes in these neuronal types were examined quantitatively by comparison of the total length and number of branches. We found that 20-hydroxyecdysone had little direct effect on the morphological development of antennal-lobe neurons, but brief exposure to sensory axons in vivo prior to dissociation significantly enhanced subsequent outgrowth in culture. PMID- 8409977 TI - Transplantation of neurons reveals processing areas and rules for synaptic connectivity in the cricket nervous system. AB - In order to assess the nature of spatial cues in determining the characteristic projection sites of sensory neurons in the CNS, we have transplanted sensory neurons of the cricket Acheta domesticus to ectopic locations. Thoracic campaniform sensilla (CS) function as proprioceptors and project to an intermediate layer of neuropil in thoracic ganglia while cercal CS transduce tactile information and project into a ventral layer in the terminal abdominal ganglion (TAG). When transplanted to ectopic locations, these afferents retain their modality-specific projection in the host ganglion and terminate in the layer of neuropil homologous to that of their ganglion of origin. Thus, thoracic CS neurons project to intermediate neuropil when transplanted to the abdomen and cercal CS neurons project to a ventral layer of neuropil when transplanted to the thorax. We conclude that CS can be separated into two classes based on their characteristic axonal projections within each segmental ganglion. We also found that the sensory neurons innervating tactile hairs project to ventral neuropil in any ganglion they encounter after transplantation. Ectopic sensory neurons can form functional synaptic connections with identified interneurons located within the host ganglia. The new contacts formed by these ectopic sensory neurons can be with normal targets, which arborize within the same layer of neuropil in each segmental ganglion, or with novel targets, which lack dendrites in the normal ganglion and are thus normally unavailable for synaptogenesis. These observations suggest that a limited set of molecular markers are utilized for cell-cell recognition in each segmentally homologous ganglion. Regenerating sensory neurons can recognize novel postsynaptic neurons if they have dendrites in the appropriate layer of neuropil. We suggest that spatial constraints produced by the segmentation and the modality-specific layering of the nervous system have a pivotal role in determining synaptic specificity. PMID- 8409978 TI - Different forms of the neurotrophin receptor trkB mRNA predominate in rat retina and optic nerve. AB - The expression of TrkB mRNAs was investigated in rat retina and optic nerve. A 11.5 kb transcript that encodes full-length TRKB was found to predominate in Northern blots of retinal RNA. By in situ hybridization, this trkB expression was concentrated in the ganglion cell and inner nuclear layers. Furthermore, an antibody to the full-length TRKB immunostained retinal ganglion cells and their axons. In contrast, Northern blots of optic nerve RNA showed a prominent 9.5 kb band that encoded a form of the TRKB receptor lacking the tyrosine kinase domain. This species was also detected in both the sciatic nerve and cultured astrocytes and C6 glioma cells. These results suggest that neurons express the full-length TRKB containing the tyrosine kinase domain, while non-neuronal cells express the truncated form of the receptor. These two classes of TRKB may mediate different neurotrophic actions in the retina and optic nerve. PMID- 8409979 TI - Differential sensitivity to androgens within a sexually dimorphic muscle of male frogs (Xenopus laevis). AB - Male frogs use their forelimb flexor muscles to clasp females during the mating behavior known as amplexus. We investigated the effects of testosterone on a principal forelimb flexor, the flexor carpi radialis muscle (FCR), using morphological and histochemical techniques. Male Xenopus laevis were surgically manipulated to produce high or low levels of circulating testosterone for an 8 week period. After this treatment, measurement of fibers in muscle cross-sections revealed that average fiber size was positively correlated with testosterone level. This effect was not the same for all muscle fibers, however. Fibers in the shoulder region were more sensitive to testosterone than fibers in other regions of the muscle. Histochemical staining of cross-sections showed that the patterns of staining for myosin ATPase or succinic dehydrogenase (SDH) were not influenced by testosterone levels, but total SDH activity was increased by testosterone treatment. When sensitivity to testosterone was correlated with ATPase activity, fibers with high ATPase activity were found to be more sensitive to testosterone than fibers with low activity, regardless of position within the muscle. Most fibers with high ATPase activity were located in the shoulder region of the muscle. These fibers are innervated by different motor axons than are fibers in the elbow region of the muscle, and contractions of shoulder (but not elbow) region fibers, elicited by stimulation of motor axons, are slowed by testosterone treatment (Regnier and Herrera, 1993, J. Physiol. 461:565-581). PMID- 8409980 TI - The quantitative relationship between olfactory axons and mitral/tufted cells in developing Xenopus with partially deafferented olfactory bulbs. AB - Partial deafferentation of the olfactory bulb in Xenopus embryos was performed to analyze the effects of afferent innervation on the development of the central olfactory structure. In an attempt to analyze a possible early inductive role of the olfactory axons, one olfactory placode was removed before differentiation of the neural tube began (stages 26-31). A morphological and quantitative analysis was performed on larvae at the onset of metamorphic climax (stage 58). When the single olfactory nerve innervated one side of the rostral telencephalon, a single olfactory bulb developed on that side and no olfactory bulb formed on the contralateral side. When the nerve innervated the midline of the rostral telencephalon, a smaller-than-normal, fused olfactory bulb developed. Partial deafferentation at these early stages resulted in a significant reduction in the number of olfactory axons (to approximately one-half of control values) and a corresponding decrease in the number of mitral/tufted cells (output neurons of the olfactory bulb). To control for possible damage to the neural tube during olfactory-placode removal, a portion of the neural tube directly beneath one of the olfactory placodes was removed in embryos. In these animals, the neural tube regenerated within 24 h and formed a normal olfactory bulb; olfactory axon and mitral/tufted-cell numbers were not significantly different from controls. In conclusion, olfactory-afferent innervation was critical for differentiation of the olfactory bulb, and decreasing the number of olfactory axons resulted in a reduction in the number of output neurons of the olfactory bulb. PMID- 8409981 TI - Modulation of the enkephalinergic phenotype of rat sympathetic neurons by hormonal and transsynaptic mechanisms. AB - Most sympathetic neurons contain one or more neuropeptides in addition to catecholamines. Although the regulation of catecholamines has been studied extensively, comparatively little is known about the regulation of neuropeptides. Since glucocorticoids and preganglionic innervation regulate catecholaminergic properties in chromaffin cells, we examined the effects of these factors on a co localized neuropeptide, leucine enkephalin (L-Enk), in adult rat sympathetic neurons in vivo. Lowered serum glucocorticoid levels as a consequence of bilateral adrenalectomy resulted in a reduction of ganglionic L-Enk content that was reversed by exposure of adrenalectomized animals to the synthetic glucocorticoid, dexamethasone. Surgical denervation of the SCG eliminated L-Enk IR preganglionic fibers and caused a dramatic increase in the number of L-Enk-IR neurons. Inhibition of the enkephalinergic component of the preganglionic innervation by chronic exposure to the opiate receptor antagonist naloxone increases the number of L-Enk-IR cell bodies and total ganglionic L-Enk content. None of the experimental manipulations noticeably altered the number or distribution of NPY-IR neurons, suggesting that the effects of glucocorticoids and the innervation on ganglionic L-Enk levels were specific and not simply an alteration of the biosynthetic state of the cells. These results demonstrate that circulating glucocorticoids and the preganglionic innervation regulate L-Enk levels in sympathetic neurons. Since both glucocorticoid levels and preganglionic activity are known to be altered by stressful stimuli, acute regulation of sympathetic L-Enk levels may constitute an important component of the autonomic response to stress. PMID- 8409982 TI - Dramatic stabilization of ferricytochrome c upon reduction. AB - By combining measurements of the free energy of denaturation of the C102T variant of Saccharomyces cerevisiae iso-1-ferricytochrome c with determination of the formal potentials for the native and chemically-denatured states we have determined the free energy of denaturation of the ferro form of the protein. We report that the simplest of all chemical modifications, addition of an electron, increases the stability of ferricytochrome c by approximately 10 kcal mol-1 at 300 K, pH 4.6. This makes reduced cytochrome c one of the most stable proteins yet investigated. PMID- 8409983 TI - Changes in global stability and local structure of cytochrome c upon substituting phenylalanine-82 with tyrosine. AB - We have examined the F82Y;C102T variant of Saccharomyces cerevisiae iso-1 cytochrome c using high-resolution proton nuclear magnetic resonance spectroscopy, chemical denaturation, and differential scanning calorimetry. Comparison of proton chemical shifts, paramagnetic shifts, and nuclear Overhauser effects indicates structural changes are localized to the vicinity of position 82. One alteration involves the rearrangement of the side chain of leucine-85. Using many more proton assignments than were available in the initial report [G. J. Pielak, R. A. Atkinson, J. Boyd, and R. J. P. Williams, Eur. J. Biochem. 177, 179-185 (1988)], a second alteration involving an interaction between arginine-13 and tyrosine-82 is observed. The interaction appears to involve a hydrogen bond with the eta-protons of arginine's guanido group acting as donor and tyrosine's phenolic eta-oxygen as acceptor. In spite of this potentially-stabilizing interaction, the free energy of denaturation decreases by approximately 2.4 kcal mol-1. Results are discussed with respect to alterations in the native and denatured states. PMID- 8409984 TI - Synthesis and antitumor activity of ammine/amine platinum(II) and (IV) complexes. AB - Dimeric platinum complexes, [Pt(RNH2)I2]2 (where R = H, methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl), have been synthesized by reactions of diiodoplatinum compounds with perchloric acid in water/ethanol solutions. The dimerization varies from several hours to a few days depending upon the length of the carbon chain in the alkylamines and the process can be conveniently monitored by 195Pt NMR spectroscopy. All these dimers exhibit two closely separated resonances around -4000 ppm (vs K2PtCl4 at -1620 ppm) in dimethylformamide. Reactions of [Pt(NH3)I2]2 with alkylamines do not yield the desired mixed ammine/amine complexes, which are obtained subsequently by treatment of the alkylamine dimer [Pt(RNH2)I2]2 with ammonium hydroxide in water. By using this latter procedure, a novel class of ammine/amine platinum complexes of the type PtII(NH3)(RNH2)Cl2, PtIV(NH3)(RNH2)X2A2, and PtIV(NH3)(RNH2)(CBDCA)A2.H2O, where X2 = chloro or 1,1-cyclobutanedicarboxylato (CBDCA), A = OH, Cl, or OCOCH3, have been synthesized and characterized by elemental analysis, infrared, and 195Pt NMR spectroscopic techniques. The alicyclic ammine/amine Pt(II) complexes, where R is C3-C6 were selected as representative of the class to undergo antitumor evaluations. The compounds had excellent activity against murine leukemic L1210/0 cells with cyclobutylamine-, cyclopentylamine- and cyclohexylamine-containing complexes demonstrating cytotoxicity superior to that of the clinically established cisplatin. PMID- 8409985 TI - High failure rate of bulk femoral head allografts in total hip acetabular reconstructions at 10 years. AB - The authors evaluated the radiographic appearance and functional performance of 30 cemented total hip arthroplasty acetabular reconstructions in 28 patients in whom bulk, weight-bearing, femoral head allografts were used to augment severe acetabular bone deficiency with a mean follow-up period of 10 years (range, 8 13.3 years). The average age of the patients was 51 years. The current study group represents the 10-year subset of a larger series of 38 hip reconstructions previously reported. The graft was bolted within the acetabulum in 12 hips and to the lateral wing of the ilium in the other 18 hips. On average, these grafts supported approximately 60% of the acetabular component. All grafts united. Failure of fixation of the acetabular component occurred in 47% of the acetabular reconstructions (14 hips in 14 patients). Four were diagnosed as loose on the basis of radiographic criteria alone and the other 10 hips had a loose acetabular component at reoperation. Loosening occurred in 58% (7 of 12) of the sockets with intraacetabular graft and in 40% (7 of 18) of those bolted to the lateral ilium. The failure rate of 47% in these reconstructions at 10 years is in sharp contrast to high success rates at less than 5 years and argues against the use of bulk weight-bearing allografts for most situations unless the only alternative is resection arthroplasty. PMID- 8409986 TI - Roentgenographic procedure for selecting proximal femur allograft for use in revision arthroplasty. AB - A roentgenographic scoring system was developed to help orthopaedic surgeons and bone banks estimate the quality of bone stock in proximal femoral allografts intended to use in revision arthroplasty. This system scores a standardized anteroposterior roentgenograph of the proximal femur using four indices representing morphological features of cancellous and cortical bone known to be clinically associated with bone strength. The indices were combined to give a weighted score, which was thought to reflect the ability of a bone to carry in vivo loads. Thirty bones were evaluated for bone mineral density using dual photon absorptiometry. They were then sent to another institution and evaluated using the newly devised roentgenographic scoring system. The results showed that the bone score roentgenographic method is a reasonable technique for selecting allograft femurs for transplantation. This roentgenographic technique and scoring system has now been packaged into kits and is available to orthopaedic surgeons and bone banks for evaluating bone stock quality in proximal femoral allografts intended for transplantation. PMID- 8409987 TI - Revision total hip arthroplasty. The use of solid allograft. AB - Intact solid allograft was used to augment severe bone loss in 8 acetabula and 15 femora in 22 patients who underwent revision total hip arthroplasty with noncemented implants. The average follow-up period was 4 years. The average time to radiographic union of whole acetabular allografts was 11 months. Superior migration of the allografts occurred in four patients. Varus tilt of the acetabular component within the allografts was noted in three patients. Eleven patients underwent entire proximal femoral allograft reconstruction, and four patients had femoral head allograft reconstruction. Nine patients with entire proximal femoral allografts achieved radiographic union at an average of 13 months and two failed by nonunion. Only one of the four patients with femoral head allograft reconstruction achieved union. Femoral component subsidence was noted in seven whole proximal femoral allografts (64%). Entire proximal femoral allograft reconstruction was complicated by at least one episode of postoperative dislocation in 6 of 11 procedures. The authors recommend that femoral head allografts should be used with caution to reconstruct proximal femoral deficiencies in which structural support is required for stability of the implant. Successful use of acetabular allografts requires that the majority of the allograft be contiguous to host bone and not to soft tissue. With entire proximal femoral reconstruction, the tip of the femoral component should not reside at the host-graft junction. All components should be cemented into allograft bone; and revision surgery should be performed before osteolytic destruction of bone advances to the point where allograft reconstruction is required. PMID- 8409988 TI - Bone-grafting for acetabular deficiency during primary and revision total hip arthroplasty. A radiographic and clinical analysis. AB - The use of bone-graft to augment the deficient acetabulum in primary and revision total hip arthroplasty (THA) is controversial. To identify factors affecting cup loosening in patients who received a bone-graft during THA, two orthopaedic surgeons retrospectively examined sequential radiographs. The surgeons also obtained independent computer measurements of hip center and cup abduction migration from preoperative, initial, and latest postoperative radiographs. Variables studied included host factors, graft factors, and technique factors. All conclusions were based on Kaplan-Meier log-rank analysis to account for differing lengths of follow-up periods among the cases. The authors report a series of 74 consecutive cases with a minimum 24-month follow-up period (mean, 40 months). All grafts appeared to unite. The clinicians found 80% stable cups, 8% possibly loose cups, and 12% (n = 9) definitely loose cups. In retrospect, technical errors were seen in six loose cups. Five revisions for loosening (6.7% of cases) were performed. Computer measurement found cup loosening in a higher percentage of cases than detected by the clinicians and did so an average of 18 months sooner. Acetabular cup loosening was associated with the American Academy of Orthopaedic Surgeons type III defects, use of allograft versus autograft, and initial cup abduction of 50 degrees or more. Kaplan-Meier survivorship analysis found 31% of cups radiographically loose and 15% revised at 5 years or more since surgery. Acetabular bone-grafting is technically demanding and should be employed when alternative reconstructions will not give a durable result. PMID- 8409989 TI - The natural history of the silent hip in bilateral atraumatic osteonecrosis. AB - The natural history of presymptomatic ischemic necrosis of the femoral head was studied in the contralateral hip of 15 patients undergoing total hip arthroplasty for avascular necrosis between 1979 and 1984. These 15 hips were retrieved from among 416 patients undergoing total arthroplasty during this time period. Presymptomatic avascular necrosis was defined according to three strict criteria that allowed the hips to be studied from an index entry point (no evidence of avascular necrosis). Follow-up examinations of each of these 15 hips from this entry point revealed that all hips eventually collapsed at a mean of 23 months after index entry point (range, 3-66 months). These data indicate that the "silent hip" is at significant risk of developing avascular necrosis and, if it becomes involved, it progresses to collapse in a high percentage of patients in a relatively short time. There was no correlation between underlying etiology and the time to collapse. PMID- 8409990 TI - Loss of hydroxyapatite coating on retrieved, total hip components. AB - The goal of this study was to examine the early retrievals of hydroxyapatite-(HA) coated hip prostheses to assess evidence of osteoconductivity, resorption of HA, and the integrity of the HA/implant bond. Six retrieved HA-coated hip prostheses (3 femoral hip stems, 3 acetabular cups) were analyzed for the amount of bone ongrowth or ingrowth of the HA-coated surface and the extent to which the coating was still present after in vivo service. The examination of these six HA-coated prostheses indicates that HA appeared to be osteoconductive. There was evidence of debonding of HA from the smooth-surfaced femoral prosthesis, although that may have been a result of the extraction process. The five plasma-spray surfaced, HA coated prostheses showed evidence of considerable loss of the HA coating at the time of receipt in the authors' laboratory, although it is was not possible to determine the cause of the loss of coating. PMID- 8409991 TI - Total knee arthroplasty in the octogenarian. AB - Fifty consecutive Miller-Galante (Zimmer, Warsaw, IN) and Kinematic II (Howmedica, Rutherford, NJ) total knee arthroplasties for osteoarthritis in patients 80 years of age and over were compared to 50 arthroplasties in patients aged 65-69 years, with a minimum 2-year follow-up period. Each patient was assessed clinically using the Hospital for Special Surgery knee rating scores and radiographically by an independent observer. Average age of the study group participants was 83 years and of the comparison group, 68 years. Apart from age, the two groups were similar with respect to sex, bilaterality, prosthetic type, and preoperative Hospital for Special Surgery scores. No significant differences were noted for pain, functional level, strength, stability, or range of motion throughout the 2-year follow-up period. More octogenarian patients were found to have a preoperative deformity, and these deformities were larger, on average, than in the retiree group, suggesting a more advanced disease pathophysiology. A larger number of elderly patients required continued use of walking aids. An analysis of the cost-effectiveness data, collected for both groups, showed that although the in-hospital costs and length of stay were the same ($17,160 Can), the cost per patient after discharge was slightly greater ($2,00 Can) in the octogenarian. This prospective clinical study of total knee arthroplasties for osteoarthritis has shown that it is a reliable and cost-effective procedure for the octogenarian. PMID- 8409992 TI - Revision hip arthroplasty. Can the octogenarian take it? AB - Fifty-six patients over the age of 80 undergoing revision hip arthroplasty in 57 hips were reviewed. The average age of the patients at the time of surgery was 81 years 6 months and the mean follow-up period was 2 years 6 months. No complications were seen in 21 patients. The complication encountered in the others were medical in 35 and procedure-related in 15. The preoperative risk factor rating (American Society of Anesthesiologists classification) correlated very well with the incidence and grading of medical complications. This correlation was significant, emphasizing the importance of strict preoperative medical and anesthetic assessment in this group of patients. The mean d'Aubigne and Postel hip score improved from 2.8, 2.2, and 3.4 to 5.6, 4.8, and 4.9 after revision surgery. PMID- 8409993 TI - Bipolar socket in protrusio acetabuli. 3-6-year study. AB - Twenty-two hips with protrusio acetabular deformity in 14 patients received bipolar socket hemiarthroplasty combined with bone-grafting to the medial wall defect. The surgical technique included crushed cancellous grafting and attention to adequate rim contact for the socket. Results were evaluated at a mean of 54 months after surgery (range, 36-76 months). On hip had thigh pain attributable to a loose femoral component. No patient had groin pain at follow-up examination. A satisfactory range of motion was achieved in all but one case, and an average increase of 40 points in the Harris hip score was realized. Radiographically, all acetabuli were returned to a more normal, inferolateral position. Socket migration (measured relative to the teardrop) in the range of 3-7 mm was noted in 5 hips; the remaining 17 hips displayed no or minimal migration of the socket (< 3 mm). The bipolar socket with crushed cancellous graft appears to be an acceptable surgical alternative in protrusio acetabuli, providing a reliable relief of pain, satisfactory functional results, and a restoration of acetabular bone stock in the majority of cases. PMID- 8409994 TI - Effect of an ultrasonic device on temperatures generated in bone and on bone cement structure. AB - Cement removal during revision arthroplasty can be a tedious, time-consuming process. The usual methods of removing cement include high-speed drills, chisels, saws and reamers, which are often associated with fracture and/or perforation of the femoral shaft. Ultrasound has been used in dentistry to remove plaque and in ophthalmology to remove cataracts and is now applied to cement removal in orthopaedic surgery. There is little data available on the effect of ultrasound on temperatures generated in bone and on its effects on the structure of bone cement. A cement mantle was constructed in the intramedullary canal in each of six 10 cm lengths of human cadaver femora. A temperature probe was then inserted into predrilled holes and temperatures generated by the ultrasonic device were recorded under a variety of conditions. In addition, a cement cylinder was microscopically evaluated after an ultrasonic tool had been inserted. Temperatures generated by the ultrasonic tool in cadaver bone were no higher than previously reported temperatures of 140 degrees C generated by high-speed drills. Furthermore, temperatures at the bone-cement interface never exceeded 60 degrees C when saline irrigation was used in conjunction with the ultrasonic tools, and were below 40 degrees C 1 minute after deactivation of the device. Microscopic examination shows that ultrasound produced local changes in the structure of bone cement converting it from a microscopically spherical interlocked material to one that appears homogeneous and granular. PMID- 8409995 TI - Radiological analysis of normal axial alignment of femur and tibia in view of total knee arthroplasty. AB - Alignment of the anatomical and mechanical axes of the lower extremities was analyzed using standardized radiographs in two series of 66 femur and 44 tibia specimens. A new anatomical axis, the distal femoral anatomical axis, was introduced. This axis corresponds to the overall femoral anatomical axis and encloses a valgus angle of 6 degrees with the mechanical axis of the femur. It allows for design of shorter intramedullary guiding instruments that will still be in congruency with true femur geometry. Comparison with computerized calculations of angles on computed tomography scans on a series of 38 bones shows a high precision of measurement on standard radiographs in neutral rotation. PMID- 8409996 TI - Socket wear assessment. A comparison of three different radiographic methods. AB - Three different methods to measure wear of total hip sockets from plain radiographs were compared with the results obtained by direct assessment of the internal deformity of 28 retrieved sockets by means of a coordinate measuring machine. The values obtained by the uni-radiographic, the duo-radiographic, and the radiographic methods based on the change of distance between the centers of the socket and the prosthetic head (CEN) all correlated significantly with the standard obtained by the coordinate measuring machine, provided the sockets were not loose. The most accurate radiographic measurements were obtained with the CEN method. A prerequisite for a reliable estimation of socket wear rate is that it be based on observations of radiographically intact sockets. PMID- 8409997 TI - Failure of uncemented polyethylene acetabular components. AB - The authors describe the mode of failure of the Ring UPM (Downs Brothers, Surrey, United Kingdom) uncemented metal-on-plastic total hip arthroplasty. Erosion of the external surface of the uncemented acetabular cup led to early loosening in eight cases. The subsequent production of large quantities of polyethylene debris caused massive granulomata, resulting in osteolysis of the acetabulum and femur. Revision surgery was difficult because of the loss of bone stock. The serious consequences of failure of this design lead to the conclusion that uncemented, nonmetal-backed polyethylene acetabular components should not be used. PMID- 8409998 TI - Posterior polyethylene wear in posterior cruciate ligament-retaining total knee arthroplasty. A case study. AB - Controversy over whether to retain the posterior cruciate ligament (PCL) during primary total knee arthroplasty continues. Both the retaining and substituting designs have their respective advantages and disadvantages. In order to capitalize on the advantages of the PCL retaining design, precise techniques in ligament balancing are required. An incorrectly tensioned PCL--too tight or too loose--may exaggerate the disadvantages of the retaining design and lead to early catastrophic failure. Three cases of early catastrophic posterior wear are presented. Although no definite documentation of excessive PCL tension is available, the authors feel that this may have had a significant role in the etiology of this wear pattern. Possible causes of incorrect PCL tension as well as intraoperative techniques to balance the PCL properly are discussed. PMID- 8409999 TI - Total knee arthroplasty in elderly patients: comparison of tibial component designs. PMID- 8410000 TI - Pregnancy in multiple sclerosis. PMID- 8410001 TI - The management of acute spinal cord compression. PMID- 8410002 TI - A prospective controlled investigation of the cognitive effects of amateur boxing. AB - Eighty six amateur boxers underwent a series of neuropsychological assessments on three occasions--pre bout, immediate post bout and follow up within two years; 31 water polo players and 47 rugby union players acted as controls. The neuropsychological tests were selected as being sensitive to subtle cognitive dysfunction and formed part of a battery of other neurological and ophthalmic assessments. No evidence of neuropsychological dysfunction due to boxing was found, either following a bout or a series of bouts at follow up. None of a range of parameters including number of previous contests, recovery from an earlier bout, number of head blows received during a bout and number of bouts between initial assessment and follow up, were found to be related to changes in cognitive functioning. PMID- 8410003 TI - Pregnancy and multiple sclerosis: a longitudinal study of 125 remittent patients. AB - The relationship between pregnancy and multiple sclerosis (MS) was assessed in a clinic-based, prospectively followed, population of 125 patients with a remittent onset of MS who had been followed for a mean (SD) of 10.3 (0.1) years. Thirty three women had a total of 49 pregnancies of which 32 had been full term and 17 terminated. There was a three-fold increase in the relapse rate per year during the first three months following delivery, compared with the baseline period of the same patients [1.62(0.38) vs 0.51(0.08) p = 0.05]. During pregnancy itself, the relapse-rate was not different from baseline. The overall relapse rate of the pregnancy group was lower than that of a control group without pregnancies after MS onset, but similar to that of patients who had children after MS onset, but no pregnancy during follow up. Pregnancy did not lead to increased disability. These results confirm that post partum increase in relapse rate is the main event related to pregnancy in MS and underline the difficulties of undertaking prospective studies in this field. PMID- 8410004 TI - Chiari malformation in adults: relation of morphological aspects to clinical features and operative outcome. AB - To determine whether clinical features attributed to cerebellar ectopia could be related to the severity of the malformation, and if morphological features could be related to operative outcome, a retrospective study of 141 patients with the adult Chiari malformation was carried out, 81 receiving operative treatment. Morphological parameters derived from preoperative clinical imaging were compared with presenting clinical features and postoperative outcomes. Patients with the most severe cerebellar malformation, defined as descent of the cerebellar tonsils to or below the axis, had disabling ataxia and nystagmus more frequently. Those with brainstem compression had limb weakness and muscle wasting more frequently. Operative outcome was significantly less favourable in patients with severe cerebellar ectopia (12% improved, 69% deteriorated) than in those with minor ectopia (50% improved, 17% deteriorated). Patients with a distended cervical syrinx had a more favourable outcome than those without. Morphological features help predict operative risk. PMID- 8410005 TI - The influence of external timing cues upon the rhythm of voluntary movements in Parkinson's disease. AB - The ability of patients with Parkinson's disease (PD) and healthy subjects to synchronise finger tapping, produced by rhythmic wrist movements, with auditory signals of target frequencies (range 1-5 Hz) and to sustain such rhythms following sudden withdrawal of auditory cues was studied. Healthy subjects were able, in the presence of auditory cues, to duplicate target frequencies accurately over the range investigated both in terms of mean tapping rate and in regularity of tapping. PD patients were less accurate under these conditions and on average tended to tap too rapidly at the lower (1-3 Hz) target frequencies and too slowly at the highest (5 Hz) target frequency. In addition, the variability of their tapping rhythms was generally greater. Healthy subjects were able to sustain tapping rhythms well following suppression of auditory signals. By contrast, withdrawal of external timing cues resulted in marked impairment of the patients' rhythm generation. At lower frequency targets (1-3 Hz) patients' tapping rates increased over rates which were already elevated in the presence of external cues. Conversely, at higher target frequencies (4-5 Hz), the average tapping rate tended to decline further from previously depressed levels. The accuracy of almost all patients fell outside the normal range. Two patterns of tapping errors were found. The first was hastening of tapping which was most evident at intermediate target frequencies. The second was faltering which occurred mainly at the higher target frequencies. These forms of behaviour may result from inherent abnormalities of internal rhythm generation since they occurred both in the presence and absence of external timing signals. Overall, our findings are consistent with the view that the basal ganglia have a role in the internal cueing of repetitive voluntary movements. PMID- 8410006 TI - Luigi Galvani (1737-98). PMID- 8410007 TI - Modulation of postural tremors at the wrist by supramaximal electrical median nerve shocks in essential tremor, Parkinson's disease and normal subjects mimicking tremor. AB - The response of postural wrist tremors to supramaximal median nerve stimulation was examined in patients with hereditary essential tremor (n = 10) and Parkinson's disease (n = 9), and in normal subjects mimicking wrist tremor (n = 8). The average frequency of on-going tremor was the same in all three groups. Supramaximal peripheral nerve shocks inhibited and then synchronised the rhythmic electromyographic (EMG) activity of all types of tremor. The duration of inhibition ranged from 90 to 210ms, varying inversely with the frequency of on going tremor. There was no significant difference in mean duration of inhibition or in the timing of the first peak after stimulation on the average rectified EMG records between the three groups. The degree to which supramaximal peripheral nerve shocks could modulate the timing of rhythmic EMG bursts in the forearm flexor muscles was also quantified by deriving a resetting index. No significant difference in mean resetting index of the three groups was found. These results suggest that such studies cannot be used to differentiate between the common causes of postural wrist tremors. PMID- 8410008 TI - Autonomic nervous system dysfunction in Parkinson's disease: relationships with age, medication, duration, and severity. AB - Heart rate variability at rest, during deep breathing, or standing up and with the Valsalva manoeuvre did not differ significantly between 67 patients with idiopathic Parkinson's disease (PD) and 31 healthy age matched controls. Blood pressure (BP) responses to standing up and sustained handgrip revealed diminished autonomic function in the PD group. In a preliminary analysis of the PD group older age, anti-Parkinson medication and higher Hoehn and Yahr (HY) stages were each associated with poor autonomic responsiveness. Disease duration was only related to the systolic BP fall on standing up. Multiple stepwise regression analysis showed that older age explained most of the variance of heart rate variability (up to 36%), and the only significant PD related factor was the use of medication, which explained less than 7%. The HY stage accounted for 12.7% of the variance in the standing up BP test, and the use of medication explained 10.6% of the variance of the systolic BP change in the sustained hand grip test. The unmedicated PD subgroup (n = 33), who had mild disease of short duration, showed no evidence of autonomic dysfunction. Cardiovascular autonomic dysfunction in PD is mild, mainly affects blood pressure responses, and occurs only in advanced cases. PMID- 8410009 TI - Cardiovascular and sweating dysfunction in patients with Holmes-Adie syndrome. AB - A cross-sectional study is reported in which 53 patients with Holmes-Adie syndrome have been subjected to a battery of tests of autonomic nervous function referable to the cardiovascular system, to two objective tests of sweating function, and to subjective assessment of sweating by application of quinizarin powder followed by body heating. The majority of patients were consecutive referrals; none was selected because of clinical indications of autonomic dysfunction. Eighty three per cent of these patients had at least one, 57% at least two, and 40% at least three objective test abnormalities, as defined by values lying outside 95 percentiles of healthy subjects who were matched for age and subjected to the same tests. In the context of multiple testing, the probability of finding outside values was such that a minimum of 3 was required to define abnormality. On this basis 40% of patients were found to have significant evidence of autonomic dysfunction. The most frequent abnormalities were impaired digital vasoconstriction to cold (23%), a reduced heart rate response to the Valsalva manoeuvre (17%), and excessive variability in sweating between test sites (in one of the tests, 43%) which is consistent with patchy loss. Abnormal quinizarin test appearances were seen in 10 patients and in a further five patients the appearances were thought to be suggestive of abnormality. Though assessment of the results of this test are subjective, the observations are consistent with the findings obtained from the objective tests which were applied. Cardiovascular and sweating abnormality did not concur significantly and only the former was found to increase progressively with known duration of the pupillotonia. It is concluded that Holmes-Adie syndrome is commonly accompanied by progressive mild but widespread autonomic involvement but rarely is this symptomatic. If symptoms suggestive of autonomic neuropathy are found in a patient with tonic pupils, a careful search for some other generalised disorder is recommended. PMID- 8410010 TI - F tacheodispersion. AB - F tacheodispersion is defined as the distribution of the conduction velocities of individual or small groups of nerve fibres estimated from significant numbers of consecutively recorded F waves. The ulnar and peroneal nerves in 18 healthy subjects were studied using this method and histograms of motor fibre conduction velocities for the control nerves were created. F tacheodispersion was applied in nine patients with neuropathies and radiculopathies selected on the basis that at least one nerve was 'normal' as measured by conventional techniques (M response, F wave minimum latency to height). In the patient group it was demonstrated that a significant proportion of motor nerve fibres had F tacheodispersion conduction velocities below normal limits despite normal conventional findings. It is concluded that F tacheodispersion should be considered in routine neurophysiological investigation when conventional methods had failed to reveal a suspected nerve lesion. PMID- 8410011 TI - Evaluation of the effect of treatment on movement disorders in astrocytomas of the basal ganglia and the thalamus. AB - Twenty patients with movement disorders associated with astrocytomas (grade I-IV according to the WHO tumour classification) of the basal ganglia and the thalamus were evaluated for the effects of treatment. Five patients had more than one movement disorder when the histological diagnosis was verified by stereotactic biopsy. Twelve had tremors, eight hemidystonia, three hemichorea, and one hemichorea/ballismus, and myoclonus respectively. Ten patients died during the follow up period, and for the surviving patients follow up periods ranged from 6 21 years. The movement disorders changed over long periods of time related to therapeutic interventions. CSF shunt operations and percutaneous radiotherapy had no definite effect on the movement disorders. There was a moderate response to medical treatment in a few patients. Stereotactic aspiration of tumour cysts had a marked influence on the movement disorder in two patients, and functional stereotactic surgery abolished tumour induced tremor in one. Interstitial radiotherapy was performed in fifteen patients for treatment of the underlying neoplasm and resulted in different and variable alterations of the movement disorders. These differences may be explained by complex interactions involving structures affected primarily by the tumour, as well as by secondary functional lesions of adjacent structures. PMID- 8410012 TI - Catecholamine levels in plasma and CSF in migraine. AB - There is clinical and pharmacological evidence of the existence of sympathetic dysfunction in migraine. Adrenaline and noradrenaline concentrations were determined in plasma and CSF of patients during attacks of common or classic migraine, comparing them with controls suffering from stress. Plasma noradrenaline levels were significantly lower in the patients with common migraine than in controls (p < 0.05). Other catecholamine levels in plasma and CSF in both migraine groups were only slightly lower than in controls. Our results suggest that central sympathetic dysfunction exists in patients with migraine. PMID- 8410013 TI - Familial progressive aphasia: its relationship to other forms of lobar atrophy. AB - Two brothers presented with slowly progressive aphasia. One brother, who became behaviourally disturbed only at the end of his illness, was found at necropsy to have predominant left frontotemporal atrophy. The other brother developed severe behavioural disturbances shortly after the onset of language impairment. His brain revealed bilateral frontotemporal atrophy. In both there was non Alzheimer's disease pathology with the histological features of loss of large cortical nerve cells, spongiform change and mild gliosis. The differential anatomical atrophy supports the view that clinical manifestations of lobar atrophy are dictated by the topographical distribution of a common underlying pathology, linking the syndromes of progressive aphasia to dementia of frontal lobe type (DFT) and DFT with motor neuron disease. PMID- 8410014 TI - Upbeat nystagmus in a patient with a small medullary infarct. AB - A 43 year old man presented with decreased sensitivity in the left side of the face and both upper limbs, and with upbeat nystagmus and skew deviation. MRI demonstrated a well defined lesion compatible with an infarct in the left side of the medulla, caudal and ventral to the vestibular nuclei, possibly involving the most caudal of the perihypoglossal nuclei, the nucleus intercalatus. Ocular motor studies, using an infrared system, showed that the nystagmus slow phase decayed exponentially, suggesting a failure of integration for vertical eye movements. Vertical integration might, therefore, be performed partly in the nucleus intercalatus. PMID- 8410015 TI - Peripheral neuropathy as the presenting feature of tyrosinaemia type I and effectively treated with an inhibitor of 4-hydroxyphenylpyruvate dioxygenase. AB - A 21 month old girl presented with a short history of frequent falls and a right sided foot drop. She went on to suffer recurrent episodes of distal weakness in her arms and legs with hyporeflexia. Electrophysiological studies were consistent with inflammatory demyelinating polyradiculoneuropathy (IDP) and treatment with corticosteroids appeared to lead to an improvement. However, the development of hypertension, evidence of tubulopathy, and hepatomegaly led to re-evaluation. A diagnosis of type I tyrosinaemia was made, based on increased urinary excretion of succinylacetone and decreased activity of fumarylacetoacetase in her cultured skin fibroblasts. A low tyrosine diet did not prevent life-threatening exacerbations of neuropathy but intravenous haemarginate appeared to aid her recovery from one exacerbation. An immediate improvement in strength was seen after starting treatment with 2-(2-nitro-4-trifluoro-methyl-benzoyl)-1,3 cyclohexanedione (NTBC), an inhibitor of 4-hydroxy-phenylpyruvate dioxygenase. A liver transplant was performed but the patient died of immediate postoperative complications. Tyrosinaemia needs to be considered in a child with recurrent peripheral neuropathy because (i) the signs of liver disease and renal tubular dysfunction may be subtle; (ii) acute exacerbations may be life threatening; (iii) specific forms of treatment are available. PMID- 8410016 TI - Guillain-Barre syndrome following jellyfish stings (Pelagia noctiluca) PMID- 8410017 TI - Recurrent Guillain-Barre syndrome following acute filariasis. PMID- 8410018 TI - Hypokalaemia mimicking Guillain-Barre syndrome. PMID- 8410019 TI - Polyneuropathy following parathion poisoning. PMID- 8410020 TI - Treatment of lower urinary tract dysfunction in patients with multiple sclerosis. European Group on SUDIMS. PMID- 8410021 TI - The use of multiple anti-dystrophin antibodies in Duchenne and Becker muscular dystrophy. PMID- 8410022 TI - Focal weakness following herpes zoster. AB - Three patients presented with focal weakness of an arm which followed segmental herpes zoster affecting the same limb. Neurophysiological investigations suggest that the site of the lesion lay at the root, plexus, or peripheral nerve level. This reflects the various ways in which the virus may affect the peripheral nervous system. PMID- 8410023 TI - Neuropathological studies of the spinal cord in early stage HTLV-I-associated myelopathy (HAM). AB - Necropsy findings for a patient with HTLV-I-associated myelopathy (HAM) of 9 months clinical duration are reported. Loss of myelin sheaths and axons together with perivascular lymphocytic infiltration was seen in the lateral and posterior columns of the spinal cord from the cervical to the lumbar region where vacuolar changes caused by the splitting of myelin sheaths were prominent. Immunohistochemical analyses revealed CD8+ cytotoxic T cell infiltration predominated in the absence of HTLV-I core protein antigen bearing-cells in the brain and spinal cord. Myelin sheath damage and predominant CD8+ cytotoxic T cell infiltration are thought to be the main neuropathological findings in the spinal cord in early stage HAM. PMID- 8410024 TI - A clinico-pathological study of adult histiocytosis X involving the brain. AB - Adult histiocytosis X involving the CNS caused progressive spastic paraparesis. The diagnosis was made by immunoreactive anti-S100 protein antibody staining and from the presence of Birbeck granules in biopsy specimens of skin lesions. Neuropathological examination showed massive proliferation and infiltration of S 100 containing histiocyte-like cells and reactive astrocytes throughout the CNS. PMID- 8410025 TI - The metabolic effects of limbic leucotomy in Gilles de la Tourette syndrome. AB - Regional cerebral oxygen metabolism was measured before and after limbic leucotomy in a patient with Gilles de la Tourette syndrome, obsessive compulsive disorder, and obsessional slowness. The preoperative scan showed hypermetabolism in the caudate nuclei, which normalised after operation. It is proposed that the beneficial effects of this operation on both tics and obsessive compulsive behaviour are mediated by disruption of abnormal neural activity in basal ganglia thalamocortical loops. PMID- 8410026 TI - Alien hand sign in association with Alzheimer's histopathology. AB - A 68 year old man is described with an alien left hand, cortical myoclonus, bilateral parietal lobe dysfunction and memory impairment but preserved language skills. The clinical diagnosis was of corticobasal degeneration but at necropsy, four years after the onset of symptoms, the pathology was of Alzheimer's disease together with some scattered chromatolytic pale neurons in the cerebral cortex. The alien hand sign has not previously been described in Alzheimer's dementia and is an illustration of the clinical heterogeneity that may occur in association with Alzheimer histopathology. PMID- 8410027 TI - Repetitive speech disorder resulting from infarcts in the paramedian thalami and midbrain. AB - A repetitive speech disorder resulting from infarcts in the paramedian thalami and the midbrain is reported. Although the speech disorder seemed like stuttering, the compulsive repetitions, constant rate and monotonous tone were not associated with ordinary stuttering. Since repetition was restricted to the first syllable, the speech disorder in our patient could be distinguished from palilalia. The extrapyramidal system is considered responsible for repetitive speech disorders resulting from infarcts in the paramedian thalami and the midbrain but without good reason. Repetitive speech disorder in patients with infarcts in the supplementary motor area (SMA) have similar clinical features to our patient. It is suggested that interruption in the projective system to the SMA is a possible cause of "stuttering like repetition". PMID- 8410028 TI - "Crossed homonymous hemianopia" and "crossed left hemispatial neglect" in a case of Marchiafava-Bignami disease. AB - "Crossed homonymous hemianopia" and "crossed left hemispatial neglect" were observed in a woman with Marchiafava-Bignami disease. Two forms of "crossed homonymous hemianopia" were observed. Initially, Goldmann perimeter testing showed a left homonymous hemianopia with the right hand and vice versa. Later, confrontation tests showed a left homonymous hemianopia, whereas visual field testing using the Goldmann perimeter (kinetic quantitative perimeter) and the OCTOPUS (Interzeag AG, static automated perimeter) showed a right homonymous hemianopia with either hand. "Crossed left hemispatial neglect" was not seen with the left hand, but neglect of the left hemifield was seen with the right hand. CT and MRI showed a lesion occupying almost the entire corpus callosum. PET showed no significant differences between comparable areas of the left and right cerebral hemispheres. These findings indicate that both signs of interhemispheric disconnection were due to the callosal lesion. Moreover, the "crossed left hemispatial neglect" can be explained as being a consequence of the dominance of the right cerebral hemisphere for visuospatial recognition. PMID- 8410029 TI - Sensory disorder of the chest as presenting symptom of lung cancer. AB - Four patients with pancoast's syndrome had burning pain in the axilla and abnormal sensation in the intercostobrachial nerve territory. The intercostobrachial nerve is the first component of the brachial plexus to be invaded by lung tumours. PMID- 8410030 TI - Some observations on the aetiology of progressive hemifacial atrophy ("Parry Romberg syndrome") PMID- 8410031 TI - Opsoclonus showing only during eye closure in hereditary cerebellar ataxia. PMID- 8410032 TI - Preferential impairment of slow alternating movements in patients with mild cerebellar ataxia. PMID- 8410033 TI - Hypersusceptibility to chemicals: risk factors for neurological disease? PMID- 8410034 TI - Philippe Pinel (1745-1826). PMID- 8410035 TI - A randomised controlled trial of domiciliary and hospital-based rehabilitation for stroke patients after discharge from hospital. AB - This study compared the functional ability and perceived health status of stroke patients treated by a domiciliary rehabilitation team or by routine hospital based services after discharge from hospital. Patients discharged from two acute and three rehabilitation hospitals in Nottingham were randomly allocated in three strata (Health Care of the Elderly, General Medical and Stroke Unit) to receive domiciliary or hospital-based care after discharge. Functional recovery was assessed by the Extended Activities of Daily Living (ADL) scale three and six months after discharge and perceived health at six months was measured by the Nottingham Health Profile. A total of 327 eligible patients of 1119 on a register of acute stroke admissions were recruited over 16 months. Overall there were no differences between the groups in their Extended ADL scores at three or six months, or their Nottingham Health Profile scores at six months. In the Stroke Unit stratum, patients treated by the domiciliary team had higher household (p = 0.02) and leisure activity (p = 0.04) scores at six months than those receiving routine care. In the Health Care of the Elderly stratum, death or a move into long-term institutional care at six months occurred less frequently in patients allocated to the routine service, about half of whom attended a geriatric day hospital. Overall there was no difference in the effectiveness of the domiciliary and hospital-based services, although younger stroke unit patients appeared to do better with home therapy while some frail elderly patients might have benefited from day hospital attendance. PMID- 8410036 TI - Complications of cerebral angiography in patients with symptomatic carotid territory ischaemia screened by carotid ultrasound. AB - After nearly 40 years, carotid endarterectomy has been shown to be of benefit to patients with symptomatic carotid territory ischaemia and greater than 70% stenosis of the relevant internal carotid artery. Cerebral angiography is performed before surgery and is not without risk. These risks must be added to those of surgery before recommending the procedure to patients. The study evaluated the local, systemic and neurological complications following digital subtraction angiography with selective catheterisation of the carotid arteries in 200 patients presenting to a cerebrovascular clinic for assessment of cerebral ischaemia. All patients had carotid ultrasound screening before angiography to screen out those with normal arteries or mild disease (less than 30% stenosis of symptomatic internal carotid artery). Complications occurred in 28 patients. There were six (3%) local, two (1%) systemic and 20 (10%) neurological complications. Seventeen neurological complications occurred within 24 hours and there were three late complications (24-72 hours). Neurological complications occurred more frequently when angiography was performed by a trainee rather than a consultant neuroradiologist (p < 0.01). The neurological complications were transient (resolved within 24 hours) in 10/200 (5%), reversible (resolved within seven days) in two (1%) and permanent in 8/200 (4%). Two patients died after a stroke and two other patients suffered a disabling stroke. At 24 hours post angiography the permanent (persisting beyond seven days) neurological complication rate was 2.5%. The incidence of total neurological complications and post angiographic strokes was higher in patients with greater than 90% stenosis of the symptomatic internal carotid artery (p < 0.001). The increased use of non invasive Doppler duplex screening will reduced the absolute number of patients put at risk of angiography, yet the rate of post angiographic complications is likely to increase as patients with severe stenosis of the symptomatic internal carotid artery are probably most at risk of complications and have most to gain from carotid endarterectomy. PMID- 8410037 TI - Prevalence of age-associated memory impairment and dementia in a rural community. AB - To obtain accurate estimates of the prevalence of age-associated memory impairment, dementia, and Alzheimer's disease, a population study was carried out in Turegano, a rural community of 1011 inhabitants in the Segovia province of Spain. The study was divided into two phases: a door to door survey of the entire population aged 40 years and over (503 persons), followed by a clinical examination of suspected cases for positive and differential diagnosis of dementia and cognitive impairment. The prevalence of age-associated memory impairment was 3.6% in individuals of 40 years and over and 7.1% in individuals of 65 years and over, whereas dementia was found in 2.6% and 5.2%, respectively. The prevalence rates of both clinical conditions increased with age. The most prevalent clinical category of dementia was dementia of Alzheimer type, which represented 1.8% and 3.8% of these two age groups. The corresponding figures for vascular dementia were 0.4% and 0.9% and for secondary dementia 0.4% and 0.5%. Age-associated memory impairment is an age-dependent disorder with a high prevalence among the elderly; some of these patients may represent an early stage of Alzheimer's disease, suggesting that the prevalence of this disorder may be higher than previously estimated. PMID- 8410038 TI - A familial disorder associated with palatal myoclonus, other brainstem signs, tetraparesis, ataxia and Rosenthal fibre formation. AB - Three siblings presented with a progressive neurological disorder beginning in the third decade of life and characterised by palatal myoclonus, nystagmus, bulbar weakness and spastic tetraparesis. There was no evidence of intellectual deterioration or seizures. CT scan showed marked brainstem atrophy in two patients and basal ganglia calcification in one. MRI scan in one showed high signal in the brainstem and periventricular region and cerebral biopsy in this patient showed myelin loss and the presence of Rosenthal fibres. A similar disease affected the siblings' mother, maternal aunt and two of the aunt's daughters, suggesting an autosomal dominant mode of transmission of what appears to be a unique genetic disorder. PMID- 8410039 TI - Motor conduction block and high titres of anti-GM1 ganglioside antibodies: pathological evidence of a motor neuropathy in a patient with lower motor neuron syndrome. AB - A patient with a progressive lower motor neuron syndrome and neurophysiological evidence of motor axon loss, multifocal proximal motor nerve conduction block, and high titres of anti-ganglioside GM1 antibodies. Neuropathological findings included a predominantly proximal motor radiculoneuropathy with multifocal IgG and IgM deposits on nerve fibres associated with a loss of spinal motor neurons. These findings support an autoimmune origin of this lower motor neuron syndrome with retrograde degeneration of spinal motor neurons and severe neurogenic muscular atrophy. PMID- 8410040 TI - Retrograde amnesia after traumatic injury of the fronto-temporal cortex. AB - An industrial manager had severe retrograde and variable but usually mild anterograde amnesia four years after a head injury. MRI showed damage of both temporal poles and the lateral portion of the right prefrontal cortex. The prefrontal and temporal cortical damage on the right side extended deeply into the white matter while the temporal cortical damage on the left side was much smaller. There was an additional left temporo-parietal lesion. The patient was of average intelligence. His attention, short term memory and learning ability were average or somewhat below average. His old memories were severely affected for the personal-episodic domain and less so for semantic remote memory abilities. Therefore an anatomical dissociation between anterograde and retrograde amnesia is possible at the anterior temporal regions, possibly interacting with the prefrontal cortex; these regions seem necessary for the retrieval of old episodic memories. PMID- 8410041 TI - Exercise performance and fatiguability in patients with chronic fatigue syndrome. AB - To examine the role of delay in recovery of peripheral muscle function following exercise in the fatigue experienced by patients with the chronic fatigue syndrome (CFS) and to examine the influence of effort perception in limiting exercise performance in these patients, a study was carried out on a group of twelve patients with chronic fatigue syndrome and 12 sex and age-matched sedentary control subjects. Symptom limited incremental cycle exercise tests including measurements of perceived exertion were performed followed by examination of the contractile properties of the quadriceps muscle group for up to 48 hours. Muscle function was assessed by percutaneous electrical stimulation and maximum voluntary contractions. Muscle function at rest and during recovery was normal in CFS patients as assessed by maximum isometric voluntary contraction, 20:50 Hz tetanic force ratio and maximum relaxation rate. Exercise duration and the relationship between heart rate and work rate during exercise were similar in both groups. CFS patients had higher perceived exertion scores in relation to heart rate during exercise representing a reduced effort sensation threshold of 3.2 units on an unmodified Borg scale in CFS patients. Patients with chronic fatigue syndrome show normal muscle physiology before and after exercise. Raised perceived exertion scores during exercise suggest that central factors are limiting exercise capacity in these patients. PMID- 8410042 TI - Creutzfeldt-Jakob disease and lyophilised dura mater grafts: report of two cases. AB - Two further cases of Creutzfeldt-Jakob disease (CJD) in association with cadaveric dura mater grafts are described. The clinical features of all such reported cases resemble more closely those of sporadic CJD, in contrast with kuru and the cases of CJD which have arisen after therapy with human pituitary-derived growth hormone. This observation may reflect the route of inoculation of the agent. PMID- 8410043 TI - Geographic variations in Wilson's disease. AB - Certain features of Wilson's disease (WD) in Asia have been found to be different from those in other continents. The higher prevalence rate in Japan is presumably due to a higher consanguinity rate. In Chinese there is a tight linkage between WD and two gene loci for esterase D and retinoblastoma in the long arm of chromosome 13. The high proportion of patients with hepatic presentation accounts for early onset of WD in the Japanese and Chinese series. Skeletal involvement, leg hyperpigmentation, dark complexion, amenorrhea, epileptic seizures, and cerebral white matter degeneration are relatively more common among WD patients in Asia. Excessive copper in the liver appears to have a protective effect against hepatocellular carcinoma and type B hepatitis. Electrophysiological studies suggest widespread functional disturbances of the CNS in WD. Side-effects from penicillamine are rather frequent and often lead to interruption of the therapy. Trien is found to be effective without adverse reactions. Oral zinc therapy may be a suitable alternative for long-term management of WD patients in developing Asian countries. PMID- 8410044 TI - Patients with chronic glucocorticoid treatment develop changes in muscle glycogen metabolism. AB - High dose glucocorticoid may induce a significant myopathy with loss of thick myofilament from muscle, particularly if administered in conjunction with depolarizing drugs. Remarkably, the effect of chronic low dose glucocorticoid in muscle is vastly different, although it may induce changes in muscle glycogen metabolism as evidenced in animal experimental trials. However, there is no clear confirmation that these changes could develop similarly in patients. We evaluate clinical, functional, histological and metabolic muscle changes during chronic low-dose glucocorticoid treatment in 11 asthmatic patients. Remarkably, these patients did not develop clinical symptoms of myopathy nor significant muscle weakness or morphological changes in muscle histology. However, glycogen concentration and the activity of the main regulatory enzymes of glycogen metabolism, aldolase and creatine kinase were modified in comparison with controls. An increase in the synthesis and muscle cell deposition of glycogen and a decrease in the muscle glycogen degradation process have been suggested. These changes were not related with malnutrition. There was not correlation between histological and biochemical changes. We conclude that chronic treatment with glucocorticoid causes clear changes in glycogen metabolism in the skeletal muscle, resulting in glycogen muscle storage. The significance of these biochemical changes is unknown, but it can be well an associated phenomenon with glucocorticoid treatment. PMID- 8410045 TI - The effects of high and low BACs on the Hoffmann reflex. AB - The purpose of this study was to measure and assess the magnitude and latency of the H-reflex and M-response between pre- and post-alcohol consumption. Also of interest was the comparison of the H-reflex and M-response between the rising and falling curves of BAC. Seven male volunteer subjects participated in this study. Testing started with pre-alcohol BAC and EMG recordings from the tibial nerve following an electrical stimulation. After alcohol consumption, BAC readings were obtained every five minutes, and the EMGs were recorded at the following BAC levels (in mg/dl): 75, 100, and falling 75. H-latency, H-amplitude, and M response were identified within the EMG signals. The analysis focused on the comparisons of these measures between each of the BAC levels and the baseline, as well as between the rising and falling BACs. Results showed that, overall, the H reflex and M-response were depressed following alcohol consumption. In addition, the depression of H-reflex appeared to be greater during the rising curve than the falling curve. PMID- 8410046 TI - HIV-1 specificity of cerebrospinal fluid and serum IgG, IgM, and IgG1-G4 antibodies in relation to clinical disease. AB - The reactivities of intrathecal and serum IgG and IgM, and IgG1-4 subclass antibodies to various HIV-1 proteins were assessed by immunoblotting at various stages of HIV-1 infection. All patients were examined neurologically including CT and/or MRI, and with HIV-1-specific and nonspecific tests of the cerebrospinal fluid (CSF). In early infection, the occurrence of anti-gag antibodies in both CSF and serum was higher than that of anti-pol antibodies among all IgG subclasses (P < 0.05). Also in late infection, anti-gag IgG1 response was most frequent (P < 0.04), while anti-gag IgG3 and IgG4 reactivities predominated over similar anti-pol antibodies (P < 0.05, respectively). Of anti-pol reactivities, in the CSF of subjects at early infection anti-p32 IgG and IgG1 antibodies were more frequent than in patients at late stages (P < 0.015). In late infection, however, the occurrence of anti-p64 IgM and IgG2-4 antibodies of both CSF and serum was higher than at early stages (P = 0.014). Regarding anti-env response, in patients with advanced infection, the CSF and serum IgG subclass reactivity against gp120 was restricted to IgG1. The CSF of individual patients with HIV encephalopathy showed a higher or similar occurrence of polyisotypic anti-gag and anti-pol IgG3 antibodies than corresponding serum. These results indicate association between declining frequency of anti-pol p32 and anti-env gp120 antibodies and severity of HIV-1 disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410047 TI - Screening of the mis-sense mutation producing the 717Val-->Ile substitution in the amyloid precursor protein in Japanese familial and sporadic Alzheimer's disease. AB - We investigated a C to T transition at base pair 2149 in the amyloid precursor protein gene in 41 Japanese cases of early-onset familial Alzheimer's disease (FAD), late-onset FAD and sporadic Alzheimer's disease (AD) by polymerase chain reaction and restriction enzyme polymorphism with BclI. Among 9 early-onset FAD patients derived from independent families, only one patient had the mis-sense mutation. Neither 5 patients with late-onset FAD nor 27 patients with sporadic AD had the mutation. Our result and the previous reports from Japan indicate that this type of mis-sense mutation is present in several cases of Japanese early onset FAD. On the other hand, our data suggest that this mutation is not a common cause of Japanese early-onset FAD. Moreover, this mutation could be absent in late-onset FAD and sporadic AD in Japan. Because the mutation has been reported to be rare in Caucasian early-onset FAD and to be absent in Caucasian late-onset FAD and sporadic AD, the situation of this mutation in Alzheimer's disease may be common beyond the ethnic background. PMID- 8410048 TI - Cytotoxicity of T cells for cerebral endothelium in multiple sclerosis. AB - We investigated the cytotoxic effect of peripheral blood T cells on cerebral endothelium in patients with MS. We examined in vitro the damage to 51Cr-labelled dissociated human brain endothelial cells produced by mitogen-stimulated T cell lines from patients with MS and controls. Endothelial targets were lysed by T lymphocytes from patients with acute relapsing MS during an exacerbation at every target-effector cell ratio tested compared with controls (P < 0.001). The percentage of endothelial targets lysed was not significantly increased by incubation with T cells from patients with acute relapsing MS in remission and chronic progressive MS, compared with that of normal subjects. Relapsing MS patients during an exacerbation had significantly higher interleukin-1 (IL-1) alpha concentrations in cultures of targets with effector cells than normal subjects (P < 0.02). Experiments of major histocompatibility complex (MHC) restricted cytotoxicity in MS demonstrated incomplete blocking of specific lysis by either anti-MHC class I or class II monoclonal antibody (mAb). These results indicate that cytotoxicity of T cells for cerebral endothelial cells may play a role in the initiation of immune response in acute relapsing MS during an exacerbation which appears to cause an increase in blood-brain barrier (BBB) permeability. PMID- 8410049 TI - Contrasting effects of immunosuppression on herpes simplex virus type I (HSV I) induced central nervous system (CNS) demyelination in mice. AB - We previously reported that lip inoculation of Herpes simplex virus type I (HSV I) in specific strains of mice would induce multifocal brain demyelination (MBD). The mechanisms mediating the development of MBD are unknown. In this study, five inbred strains of mice (C57BL/6J, Balb/cByJ, A/J, SJL/J, PL/J) immunosuppressed with either irradiation (IR), cyclophosphamide (CY), or cyclosporin A (CP) along with three immune deficient strains (C57BL/6J nu/nu, Balb/cByJ nu/nu, C57BL/6J bg/bg) were lip inoculated with HSV I to determine the effect of immunosuppression on viral spread throughout the brain and the development of demyelination during the acute stage of infection. Mortality increased in all groups when compared with controls but was greatest in A/J, SJL/J, and PL/J strains, where all mice died before day 6 PI. In contrast with immunocompetent C57BL/6J mice where virus is restricted to the brainstem, virus spread throughout the brain of immunosuppressed C57BL/6J, C57BL/6J nu/nu, and C57BL/6J bg/bg mice. Despite viral spread throughout the brain of immunosuppressed C57BL/6J, C57BL/6J nu/nu, Balb/cByJ and Balb/cByJ nu/nu mice, MBD did not develop. MBD did develop however, in both HSV I infected C57BL/6J bg/bg and CP treated Balb/cByJ mice. Immunosuppression of HSV I infected Balb/cByJ mice prevents the development of demyelination at the trigeminal root entry zone (TREZ) of the brainstem while in Balb/cByJ nu/nu mice, the extent of demyelination at TREZ was reduced and delayed when compared with immunocompetent controls. These results suggest that the immune system plays an important role in limiting viral spread in the brain as well as in the development of demyelination at TREZ and of MBD throughout the brain during the acute phase of infection. Virus alone does not induce MBD in this animal model of virus induced CNS demyelination but is a prerequisite for its development. PMID- 8410050 TI - Immunocytochemical studies on the vimentin distribution and cell proliferation of fibroblasts in patients with Friedreich's ataxia. AB - Fibroblasts obtained from patients with Friedreich's ataxia and normal control subjects were studied by immunocytochemistry for intermediate filament vimentin and also for in vitro proliferation. Trypsinized cells were seeded on coverslips and incubated for 1.5 h and 24 h. The expression of vimentin in cells was investigated by immunofluorescence microscopy. Cell proliferation was studied with BrdU antibody technique. Cells from patients with Friedreich's ataxia showed a slower outgrowth of vimentin filaments in comparison to cells from normal controls. These cells also incorporated less 5-bromo-2'-deoxyuridine (BrdU) into their DNA. The observations may be relevant to the clinical manifestations of the disease which involves many organs in addition to brain and spinal cord. PMID- 8410051 TI - Ambulatory monitoring of tremor and other movements before and after thalamotomy: a new quantitative technique. AB - Tremor, e.g. in Parkinson patients, often shows large spontaneous fluctuations in severity over the day, to such an extent that a short observation is usually not sufficient to assess the overall severity or the effect of a treatment. Since momentary impressions of the tremor can thus be misleading, long-term ambulatory recordings would be helpful in the evaluation of severity and treatment effectiveness. As existing methods for long-term tremor registration have several shortcomings, a new method is proposed: an algorithm was designed to discriminate tremor from other movements and to describe the amount (i.e. the proportion of tremor or movements per time unit) as well as the intensity (i.e. average acceleration amplitude) of the two types of movement. In the evaluation of the severity of tremor both the amount and intensity of tremor episodes are of importance. The algorithm was tested on 24-h analog tape recordings of wrist movement in 10 young and 10 aged controls, as well as in 8 patients with tremor- both before and after a tremor relieving thalamotomy. The algorithm scored movements as 'tremor' exclusively in patients prior to the operation. Fluctuations in tremor severity over the day were detected, and tremor could be discriminated from non-pathological movements. Moreover, following thalamotomy, motor slowing (bradykinesia) was detectable using this algorithm. Based on these test results, a miniaturized device in wrist-watch format is now being developed for long-term registrations. PMID- 8410052 TI - Immunohistochemical studies with antibodies to neurofilament proteins on axonal damage in experimental focal lesions in rat. AB - Immunohistochemistry with monoclonal antibodies against neurofilament (NF) proteins of middle and high molecular weight class, NF-M and NF-H, was used to study axonal injury in the borderzone of focal lesions in rats. Focal injury in the cortex was produced by infusion of lactate at acid pH or by stab caused by needle insertion. Infarcts in substantia nigra pars reticulata were evoked by prolonged pilocarpine-induced status epilepticus. Immunohistochemical staining for NFs showed characteristic terminal clubs of axons in the borderzone of lesions. Differences in the labelling pattern occurred with different antibodies which apparently depended on molecular weight class of NFs and phosphorylation state. These immunohistochemical changes of NFs can serve as a marker for axonal damage in various experimental traumatic or ischemic lesions. PMID- 8410053 TI - Pre-and post-mortem MR imaging of unsuspected multiple sclerosis in a patient with Alzheimer's disease. AB - A patient with the clinical diagnosis of Alzheimer's disease is presented in whom pre-mortem T2-weighted MRI revealed a periventricular white matter lesion. Postmortem T2 weighted MRIs of the formalin fixed brain revealed the same white matter lesion. Microscopically, classical Alzheimer changes were found and, unsuspectedly, the histopathological correlate of the white matter lesion proved to be an old, inactive, MS plaque. A similar lesion was discovered in the cervical myelum. These findings illustrate that T2-weighted post-mortem MRIs are highly comparable to pre-mortem images and that MRI is sensitive in detecting clinically silent white matter lesions. The histopathology of such lesions may also include MS plaques. PMID- 8410054 TI - Primary progressive versus relapsing remitting multiple sclerosis in Japanese patients: a combined clinical, magnetic resonance imaging and multimodality evoked potential study. AB - Thirty-five Japanese patients with relapsing remitting multiple sclerosis (RRMS) and 11 Japanese patients with primary progressive MS (PPMS) were compared by a combined clinical, magnetic resonance imaging (MRI) and multimodality evoked potential (MEP) study. Patients with PPMS showed a more common occurrence of gait disturbance as the initial symptoms as well as a more common occurrence of cerebellar symptomatology than did those with RRMS, while spinal cord symptomatology was frequently observed in both conditions. On brain MRI, patients with PPMS had 3 times more lesions than did those with RRMS (P < 0.001, chi 2 test). The percentage of infratentorial lesions was also significantly higher in PPMS than in RRMS on MRI. Moreover, we found a significantly higher frequency of abnormal records in visual, brainstem auditory and somatosensory evoked potentials in PPMS than in RRMS. Interestingly, clinically unexpected abnormalities were significantly more common in PPMS than in RRMS throughout all modalities of MEPs. Thus, in Eastern MS, there exists a distinction between PPMS and RRMS in the MRI and MEP findings as well as in the clinical findings. Our result therefore suggest that there may be two distinct subtypes in MS. PMID- 8410055 TI - Immunoglobulin G subclasses in older persons with Down syndrome. AB - IgG subclasses were measured in sera from 33 persons with Down syndrome (DS) (mean age 55 +/- 7 years) and 33 age- and sex-matched control individuals using a mouse monoclonal antibody based sandwich enzyme linked immunosorbent assay. Significantly higher levels of IgG1 and IgG3 and lower levels of IgG2 and IgG4 subclasses were found in the DS group compared to the control individuals. The higher levels of IgG1 and IgG3 subclasses found in DS persons were consistent with those seen in patients with autoimmune diseases and chronic viral infections; the lower levels of IgG2 and IgG4 subclasses were consistent with those seen in patients with recurrent infections. Our findings are similar to those reported in children with DS. We speculate that the subclass levels may have little or no relationship to the development of brain lesions typical of Alzheimer disease in older persons with DS. There were no significant differences between the levels of IgG subclasses of persons with DS showing signs of dementia of the Alzheimer type compared to those without such manifestations. PMID- 8410056 TI - Serum prolactin levels in active multiple sclerosis and during cyclosporin treatment. AB - Prolactin is essential for immune function. Excess prolactin augments some immune reactions, whereas low serum levels of prolactin inhibit immune function and prevent experimental allergic encephalomyelitis, an animal model of multiple sclerosis (MS). Activated lymphocytes, characteristics of MS, release prolactin. In this study, serum prolactin levels were normal in 35 patients with chronic progressive MS and 19 patients with acute exacerbations. These results suggest it is unlikely that prolactin contributes to the enhanced immune reactivity characteristic of MS. Acute cyclosporin A (CsA) administration increases circulating prolactin levels in animals and might paradoxically augment some immune reactions. We find that chronic CsA therapy for MS does not cause elevations in serum prolactin and should not reverse any therapeutic effect of CsA. Disturbances of prolactin regulation are not characteristic of MS. PMID- 8410057 TI - Reactivity of neuroborreliosis patients (Lyme disease) to cardiolipin and gangliosides. AB - A subset of patients (50%) with neuroborreliosis (Lyme disease) showed IgG reactivity to cardiolipin in solid phase ELISA. In addition, a subset of patients with neuroborreliosis (29%) and syphilis (59%) had IgM reactivity to gangliosides with a Gal(beta 1-3) GalNac terminal sequence (GM1, GD1b, and asialo GM1). Anti ganglioside IgM antibodies were significantly more frequent in these two groups of patients compared to patients with cutaneous and articular Lyme disease, primary antiphospholipid syndrome, systemic lupus erythematosus and normal controls. Correlative evidence and adsorption experiments indicated that antibodies to cardiolipin had separate specificities from those directed against the gangliosides. IgM antibodies to Gal(beta 1-3) GalNac gangliosides appeared to have similar specificities since these were positively correlated and inhibitable by cross adsorption assays. Given the clinical associations of patients with neuroborreliosis and syphilis with IgM reactivity to gangliosides sharing the Gal(beta 1-3) GalNac terminus, we suggest that these antibodies could represent a response to injury in neurological disease or a cross reactive event caused by spirochetes. PMID- 8410058 TI - Posterior uveitis, paraneoplastic encephalomyelitis and auto-antibodies reacting with developmental protein of brain and retina. AB - We report a 68-year-old woman who presented with posterior uveitis, cerebellar ataxia, dysautonomia, peripheral neuropathy and undifferentiated carcinoma. Autopsy showed slight brain inflammatory changes and severe Purkinje cell degeneration. By immunocytochemistry, the serum and CSF contained antibodies that reacted with the cytoplasm of cells in the brain and retina of rat. Some neurons and glial cells were stained. However, the most labelled cells were immature cells of the subventricular layers of the dentate gyrus, olfactory bulb and lateral ventricles. In rat retina, the plexiform layers and cells of the inner granular layer were stained. E15 rat embryo showed diffuse staining of post mitotic cells and process in the central and peripheral nervous system. With the patient's IgG, various bands of 45, 60, 70 and 135 kDa apparent molecular weight were demonstrated on immunoblots of adult human, adult and E15 embryonic rat brain and rat and bovine retina. Eluates of tissue bound IgG labelled the same bands. PMID- 8410059 TI - Brain embolic phenomena associated with cardiopulmonary bypass. AB - Various biologic and non-biologic materials may be embolized to the brain after the use of cardiopulmonary bypass (CPB) pumps during open heart surgery but their relative frequency and importance are uncertain. Among the nonbiologic materials, Antifoam A, which contains organosilicates and silicon, continues to be employed as an additive to prevent frothing. Recent improvements in filtration and oxygenation techniques have clearly reduced the incidence of large emboli and complications like stroke but other neurologic sequelae following open heart surgery are common and in many cases poorly explained. A recently developed histochemical technique for the demonstration of the endothelial alkaline phosphatase (AP) was employed in a post-mortem study of brains from 8 patients and 6 dogs dying within a few days after open heart surgery employing cardiopulmonary bypass perfusion. Brains from 38 patients and 6 dogs who were not subjected to heart surgery were studied as controls with the same technique. The AP-stained slides are suitable for both light microscopic examination of the thick celloidin sections as well as a subsequent processing for high-resolution microradiography. Small capillary and arteriolar dilatations (SCADs) were seen in the test subjects/animals but not controls. SCADs were seen in all parts of the brain. Approximately 50% of the SCADs showed birefringence when examined with polarized light. SCADs are putative embolic phenomena and the exact nature and source of the embolic material is under investigation. A glycolipid component is indicated by preliminary studies. SCADs are difficult to find in routine paraffin sections and most if not all of the offending material seems to be dissolved during processing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410060 TI - Right proatlantal artery type I, right internal carotid occlusion, and left internal carotid stenosis: case report and review of the literature. AB - A 66-year-old man presented with right cerebellar infarction and ischemic lesions in the left dorsal thalamus and right upper parietal lobe. Angiography showed occlusion of the right internal carotid artery proximal to an ipsilateral proatlantal artery type I, 70% stenosis of the left internal carotid artery, and aplasia of both posterior communicating arteries. The carotid occlusion was successfully treated by thrombendarterectomy. Persistence of a proatlantal artery is a rare condition. In relation to the 38 literature reports on proatlantal arteries, this case demonstrates the clinical significance of a persistent proatlantal artery in the evolution of atypical ischemic cerebrovascular disease. PMID- 8410061 TI - Multiple deletions of mitochondrial DNA in a patient with periodic attacks of paralysis. AB - In this study multiple deletions of mitochondrial genome were found in a patient presenting with periodic attacks of paralysis. Morphological studies revealed mitochondrial abnormalities along with typical histopathological features of periodic paralysis. Southern blot and PCR analysis revealed multiple mtDNA deletions. Our patient could be affected by two unrelated diseases, idiopathic periodic paralysis and presymptomatic mitochondrial myopathy. Alternatively, mtDNA alterations and oxidative deficiency might express themselves phenotypically as periodic paralytic attacks, although this correlation has never been reported. PMID- 8410062 TI - Duration dependent post-ischemic hypothermia alleviates cortical damage after transient middle cerebral artery occlusion in the rat. AB - We investigated the effect of post-ischemic 30 degrees C hypothermia on transient middle cerebral artery (MCA) occlusion in the rat. Male Wistar rats (n = 27) were subjected MCA occlusion for 2 h by inserting a nylon filament into the internal carotid artery. Three groups of animals were studied: 1) normothermic ischemia and normothermic reperfusion (n = 13), 2) normothermic ischemia and 1 h of hypothermic reperfusion (n = 6), 3) normothermic ischemia and 3 h of hypothermic reperfusion (n = 6); monitoring of cerebral temperatures were performed on two additional rats. The animals were sacrificed after one week, and coronal sections were obtained and stained with hematoxylin and eosin (H/E) for histopathological examination and determination of infarct volume. The data indicate that both normothermic reperfusion and 1 h hypothermic reperfusion groups exhibited similar infarct volumes in the cortex and the basal ganglia, respectively (P > 0.1). The 3 h post-ischemic hypothermia group revealed a significant decrease in infarct volume in the cortex compared to the normothermic group (P < 0.05). However, the infarct volume of the basal ganglia was not significantly lessened by the 3 h post-ischemic hypothermia. Thus 3 h post-ischemic hypothermia provides preferential reduction of cell damage in the cortex, from 2 h of MCA occlusion in the rat. PMID- 8410063 TI - Effects of smoking in patients with early-onset Parkinson's disease. AB - Smoking a cigarette relieved symptoms in 6 patients with early-onset Parkinson's disease. In these patients smoking reduced tremor, rigidity, bradykinesia, and gait disturbance including frozen gait. These effects lasted for about 10-30 min, and relieved parkinsonian symptoms in the off-period. Nicotine chewing gum had a lesser effect. Nicotine is thought to activate the nigrostriatal dopaminergic pathway and increase the release of dopamine in the striatum, and this can explain the effects of smoking in these patients. PMID- 8410064 TI - Neuronal cell loss of the striatonigral system in multiple system atrophy. AB - We investigated the longitudinal as well as lateral loss of striatal and nigral cells and its distribution in 7 cases of multiple system atrophy. Loss of striatal small cells and nigral pigmented cells was more prominent in the caudal part than in the rostral and mid-parts. Cell loss was especially high in the dorsolateral zone of the caudal putamen and in the lateral zone of the caudal nigra. These findings indicate that MSA predominantly disturbs the striatal and nigral efferent systems, which interlink the caudal and dorsolateral putamen with the caudolateral nigra. In less severe cases, the rostral to mid-parts of the putamen or substantia nigra were almost intact while its caudal portion was clearly affected. The degenerative process of MSA seems to occur initially in the caudal parts of the putamen and substantia nigra, extending later to the rostral parts. Thus striatal small cells and nigral pigmented cells degenerate according to anatomical relationship. In MSA, degeneration of the striatonigral system could well be explained as being transsynaptic. PMID- 8410065 TI - Whole blood monoamine oxidase activity in Parkinson's disease and multiple system atrophy patients. AB - Monoamine oxidase type B (MAO-B), which catalyses the breakdown of dopamine (DA) in human brain, is said to be involved in the pathophysiology of Parkinson's disease (PD). Activity of MAO-B in PD has been measured in platelets isolated from blood samples in different studies, with contradictory results, possibly due to the differences in substrate used or to differences in platelet isolation. Therefore we measured MAO activity in whole blood, which is almost identical to MAO-B activity in platelets, in 25 drug-naive PD patients, 25 treated PD patients, 9 multiple system atrophy (MSA) patients and 20 controls, using a spectrofluorimetric method with kynuramin as a substrate. No statistically significant differences between groups were found, nor any correlation with the severity or duration of the disease. PMID- 8410066 TI - A new analysis of mortality from motor neurone disease in Japan, 1950-1990: rise and fall in the postwar years. AB - Recent studies have demonstrated a worldwide rise in mortality from motor neurone disease (MND). However, in Japan mortality appears to have fallen significantly since the late 1960s, especially amongst women. Studies of the cause of both the worldwide rise, and the unique decline in MND mortality in Japan, have largely failed to substantiate the role of any single factor, or group of factors in these phenomena. Modelling the relationship between age and mortality using gompertzian analysis has already shown that the rise in MND mortality in England and Wales, and the United States, is mainly the result of increased longevity and decreasing competition from other causes of death amongst a susceptible subpopulation. Employing the same techniques on Japanese mortality data from 1950 to 1990 demonstrates that an unusual and accelerated increase of mortality occurred in the 1950s and 1960s, probably caused by an earlier unknown but extremely potent environmental agent or agents. This premature depletion of the susceptible subpopulation resulted subsequently in a lower than expected mortality rate. Mortality is now rising slowly to expected levels as the size of the susceptible subpopulation recovers to reach the ages at which MND is normally expressed. Further substantial rises in mortality are anticipated in future decades. PMID- 8410067 TI - Treatment of spasticity in children with low dose benzodiazepine. AB - In an attempt to investigate whether benzodiazepines at low dosage have a significant effect in reducing spasticity among children with cerebral palsy, we carried out a double-blind, placebo-controlled, cross-over study. Twelve children with either spastic diplegia or hemiplegia participated in this study. The mean age was 14 years. The restraint of passive knee movements was determined with a dynamic dynamometer and spastic stretch reflexes were measured as EMG activity in muscles stretched. Clonazepam was given at low dosage (0.02 mg/kg body weight). In each child measurements of passive restraint were made on 2 different days immediately before and 3 h after an i.m. injection of either clonazepam or placebo in randomized order. Clonazepam significantly reduced spastic restraint (P < 0.001) compared to non-significant reduction with placebo. The mean plasma concentration of clonazepam at time of spasticity evaluation was 21 mmol/l which is in the low dose range, far below conventional doses. The study thus shows a positive effect of low dose clonazepam in reducing spasticity in children when given as a single dose. PMID- 8410068 TI - Nodal and terminal sprouting by regenerating nerve in vitamin E-deficient rats. AB - The increased number of poly-innervated cells in normal and reinnervated extensor digitorum longus (edl) muscle of vitamin E-deficient rats suggests enhanced sprouting by motor neurons in conditions of decreased protection against lipid peroxidation. End-plates and terminal axons were observed by a combined technique that shows both end-plate acetylcholinesterase area and axons. Quantitative observations of nodal and terminal sprouting in normally innervated and reinnervated edl muscles of vitamin E-deficient rats were carried out. Branch points of nerve terminal within end-plates were also observed. Three main results were obtained. First, a notable increase of both terminal and nodal sprouting was found in reinnervated muscles of normal and vitamin E-deficient rats; moreover, a relative increase in the number of nodal sprouts occurs in the long run. Second, in muscles of uninjured, vitamin E-deficient rats, nodal and terminal sprouting and branching within end-plate was greater than in controls. Third, nodal sprouting by regenerating axons was more affected by vitamin E-deficiency than terminal sprouting and branching within end-plates. PMID- 8410069 TI - Photochemically induced experimental ischemic neuropathy: a clinical, electrophysiological and immunohistochemical study. AB - A new experimental model of focal peripheral nerve infarction is presented. Ischemia was produced in 12 rats by intravascular thrombosis induced by the photochemical reaction of systemically injected rose bengal to the local application of light from a cold light source. Clinical, electrophysiological and immunohistochemical techniques were used to monitor the pathology and the time course of experimental ischemic neuropathy (EIN) of the sciatic nerve. Primary axonal neurofilament disintegration was detectable 4-24 h after illumination and was followed by wallerian degeneration within the first week. At 7 days, there was a secondary disruption of myelin sheaths accompanied by massive infiltration of macrophages and phagocytosis of the necrotic debris. The majority of detected macrophages were derived from circulating blood monocytes which had invaded the nerve. Two weeks after the initial lesions, degeneration had advanced without any signs of regeneration or remyelination. Electrophysiological recordings corroborate the findings of primary axonal degeneration and failure of regeneration up to 2 weeks after the lesion. PMID- 8410070 TI - Subclinical phenotypic expressions in heterozygous females of X-linked recessive bulbospinal neuronopathy. AB - Four of 8 definite heterozygous female carriers determined by PCR amplification of tandem CAG repeat of the AR gene, from 4 families of X-linked recessive bulbospinal neuronopathy (X-BSNP) showed extensive high amplitude motor unit potentials in examined muscles although all subjects were neurologically normal. Plasma creatine kinase, myoglobin, myosin light chain, lactate and pyruvate were all normal even in the carriers who showed EMG abnormalities. Muscle biopsy showed a type 2 fiber preponderance and possible very mild type 2 fiber grouping in a carrier with an EMG abnormality. These results suggest that a mutant AR gene may express subclinical phenotypic manifestations in a subpopulation of the heterozygous females of X-BSNP. PMID- 8410071 TI - A model for monitoring nerve blood flow during expansion by laser Doppler flowmetry in the rabbit. AB - A new model is described for monitoring nerve blood flow during expansion by laser Doppler flowmetry. Rabbit sciatic nerve is gradually expanded with a custom made spherical expander, while nerve blood flow is monitored by laser Doppler flow output. This model provides a valid method of controlling nerve blood flow during expansion. PMID- 8410072 TI - Oxidative polymorphism of debrisoquine is not related to the risk of Alzheimer's disease. AB - Oxidative polymorphism of debrisoquine has been studied in patients suffering from many spontaneous disorders which show genetic and/or environmental factors in their pathogenesis. To elucidate whether any relationship exists between this genetic polymorphism and the risk of developing Alzheimer disease (AD) we determined the oxidative phenotype and metabolic ratio (MR) of debrisoquine (DBQ) in 47 patients with AD or senile dementia of Alzheimer type (SDAT) and 837 healthy controls. The patients were free of drugs during at least the previous 30 days; all the controls were free of drugs. Three patients (6.38%) and 42 controls (5.02%) were classified as poor metabolizers (PM) of DBQ (non-significant difference). The distribution of MR values in the AD/SDAT patients showed non significant differences when compared with controls. There was no relation between oxidative polymorphism of DBQ and age at onset of the disease. These results suggest that DBQ oxidative genetic polymorphism cannot be considered as a risk factor for developing AD-SDAT. PMID- 8410073 TI - Phenotypic expression of mtDNA heteroplasmy in the skeletal muscle of patients with oculomyopathy: defect in mitochondrial protein synthesis. AB - The biochemical consequences of mtDNA heteroplasmy, observed in patients with a range of diseases associated with the mitochondrial respiratory enzymes deficiency is of particular interest, as they might provide information with regard to the regulatory interactions which govern the expression of the human mitochondrial genome. Three patients with chronic progressive external ophthalmoplegia (CPEO) were investigated to study the consequences of mtDNA heteroplasmy on mitochondrial protein synthesis. All 3 patients exhibited partially deleted mtDNA species (varying in size from 10.5 to 14 kb) in their skeletal muscle, which co-existed with the normal 16.5 kb mtDNA. The examination of mitochondrial translation products following the incorporation of [35S]methionine by isolated mitochondria, showed grossly abnormal patterns of mitochondrial translation products, suggesting a major disturbance in the regulation of mitochondrial protein synthesis. PMID- 8410074 TI - Effect of sera from myasthenia gravis patients and of alpha-bungarotoxin on acetylcholinesterase during in vitro neuromuscular synaptogenesis. AB - Myasthenia gravis (MG) is mediated by circulating antibodies directed against acetylcholine receptor (AChR) but the antibody titre is poorly correlated with the clinical severity of the disease. We analysed acetylcholinesterase (AChE) activity, molecular forms and distribution during in vitro synaptogenesis, in the presence of sera from MG patient. We observed that the formation of AChE patches is inhibited in proportion to the anti-AChR antibody titre, whatever the clinical severity of the disease. The total activity and the proportion of the different molecular forms were unchanged suggesting that AChE level and distribution are controlled by independent mechanisms. To clarify the relationship between the mechanisms of AChE concentration during synaptogenesis and AChR concentration, we compared the effect of MG sera (receptors are internalised and degraded) and of the acetylcholine antagonist alpha-bungarotoxin (non-functional receptors are still present in the muscular membrane). In the presence of alpha-bungarotoxin, the number of AChR clusters, and AChE activity and concentration were equivalent to control values. The comparison of the results obtained with antibodies and alpha-bungarotoxin suggests that the presence and/or concentration of AChR is a necessary condition for normal concentration of AChE during synaptogenesis. PMID- 8410075 TI - Origins of synaptic inputs to calretinin immunoreactive neurons in the guinea-pig small intestine. AB - Calretinin immunoreactivity is almost completely confined to two classes of neuron in the myenteric plexus of the guinea-pig small intestine, longitudinal muscle motor neurons and ascending interneurons. Nerve cell bodies of the two classes can be readily identified by their sizes and positions in ganglia. The motor neurons, which are small Dogiel type I neurons, are about 20% and the interneurons, which are medium-sized Dogiel type I neurons, are about 5% of myenteric neurons. In the present work, we have also discovered a minor population (0.1%) of small filamentous neurons. In unoperated regions of intestine, at the light microscopic level, numerous calretinin immunoreactive nerve fibres were found in the tertiary plexus that innervates the longitudinal muscle and a medium density of varicose fibres formed pericellular endings in the myenteric ganglia. After double myotomy operations, in areas of plexus 0.5 to 1.5 mm wide which were isolated from ascending and descending inputs, calretinin immunoreactive fibres of the tertiary plexus were unchanged, but the pericellular endings in the ganglia disappeared. Both the ascending interneurons and the longitudinal muscle motor neurons received ultrastructurally identified synapses and close axonal contacts that were calretinin-immunoreactive. These were counted in semi-serial sections from normal intestine and from regions between myotomy operations. In unoperated intestine, the proportions of calretinin-immunoreactive synapses on small, calretinin-immunoreactive, Dogiel type I nerve cells and small filamentous nerve cells were 30% and 0.1% respectively and on medium-sized Dogiel type I cells the proportion was 28%. Electron microscopy revealed an almost complete loss of immunoreactive inputs to the small Dogiel type I cells between double myotomies, but the number of unreactive inputs was the same as in normal intestine. This work demonstrates that the ascending calretinin-immunoreactive interneurons connect with one another to form ascending chains in the myenteric plexus and that they also provide about 1/3 of the inputs received by calretinin immunoreactive longitudinal muscle motor neurons. Many of the remaining inputs to these motor neurons are local; we have deduced that these are mainly from primary sensory neurons. PMID- 8410076 TI - GABA-immunoreactivity in processes presynaptic to the terminals of afferents from a locust leg proprioceptor. AB - Individually labelled sensory neurons from the femoral chordotonal organ, a proprioceptor at the femoro-tibial joint of a locust hindleg, were analysed by intracellular recording, and by electron microscopical immunocytochemistry to reveal the arrangement of their input and output synapses and to determine whether the input synapses were GABAergic. Intracellular recordings from these sensory neurons show spikes superimposed on a barrage of synaptic potentials during movements of the femoro-tibial joint. These synaptic inputs can be mimicked by GABA. Input synapses are made onto the vesicle-containing terminals of afferents and are often closely associated with the output synapses. By contrast, the axons of the afferents in the neuropil have no vesicles and neither make nor receive synapses. The input synapses to the afferent terminals are made from processes typically a few microns in diameter, whereas the output synapses are made onto much smaller processes of only 0.1-0.2 micron. Input synapses at which an afferent terminal is the only postsynaptic element are common. Where the synapse is dyadic the second postsynaptic element does not usually appear to be a chordotonal afferent. The output synapses from the afferent terminals are usually dyadic. At 78% of the input synapses, the presynaptic neurite showed immunoreactivity to a GABA antibody, supporting the physiological evidence that the presynaptic effects can be mediated by the release of GABA. The remaining (22%) immunonegative synapses are intermingled with those showing GABA immunoreactivity, but their putative transmitter is unknown. These morphological observations suggest that the presynaptic control of the chordotonal afferents is largely mediated by GABAergic neurons, but because other types of neuron also appear to be involved, presynaptic modulation may be more complex than has yet been revealed by the physiology. PMID- 8410077 TI - Immunotyping of radial glia and their glial derivatives during development of the rat spinal cord. AB - The differentiation of glia in the central nervous system is not well understood. A major problem is the absence of an objective identification system for involved cells, particularly the early-appearing radial glia. The intermediate filament structural proteins vimentin and glial fibrillary acidic protein have been used to define the early and late stages, respectively, of astrocyte development. However, because of the non-specificity of vimentin and the temporal overlap in expression patterns of both proteins, it is difficult to refine our view of the process. This is especially true of the early differentiation events involving radial glia. Using the developmentally-expressed intermediate filament-associated protein IFAP-70/280 kD in conjunction with vimentin and glial fibrillary acidic protein markers, a comprehensive investigation of this problem was undertaken using immunofluorescence microscopy of developing rat spinal cord (E13-P28 plus adult). The phenotypes of the cells were defined on the basis of their immunologic composition with respect to IFAP-70/280 kD (I), vimentin (V) and GFAP (G). A definitive immunotype for radial glia was established, viz, I+/V+/G-; thus reliance upon strictly morphological criteria for this early developmental cell was no longer necessary. Based upon the immunotypes of the cells involved, four major stages of macroglial development were delineated: (1) radial glia (I+/V+/G ); (2) macroglial progenitors (I+/V+/G+); (3) immature macroglia (I-/V+/G+); and (4) mature astrocytes (I-/V+/G+ primarily in white matter and I-/V-/G+, the predominant type in gray matter). It is of interest to note that the cells of the floor plate were distinguished from radial glia by their lack of IFAP-70/280 kD immunoreactivity. Introduction of the IFAP-70/280 kD marker has therefore provided a more refined interpretation of the various differentiation stages from radial glia to mature astrocytes. PMID- 8410078 TI - Dense accumulations of synaptic-like microvesicles in 'dark' pinealocytes of the gerbil pineal gland. AB - In an electron-microscopical study the occurrence and ultrastructural features of electron-dense 'dark' variants of pinealocytes were evaluated in the gerbil pineal gland. A few 'dark' pinealocytes, which tended to form small clusters of contiguous cells, could consistently be detected in pineals fixed and embedded by various procedures. Apart from the different degree of electron density, the only conspicuous difference between 'dark' and electron-lucent 'light' pinealocytes concerned their compartment of synaptic-like microvesicles. Thus, both variants of pinealocytes contained abundant clear microvesicles of variable size which accumulated in dilated process terminals. However, the vesicles within the process endings of 'dark' pinealocytes showed an unusually dense arrangement throughout the cytoplasm. As was demonstrated by immunogold staining, the accumulations of vesicles in the 'dark' terminals contained synaptophysin, a major synaptic vesicle-associated protein. This protein is present in small clear vesicles with putative secretory functions in a wide variety of neuroendocrine cells and has previously been shown to be a common constituent of microvesicles in mammalian pinealocytes. Since gerbil pinealocytes displayed distinct gradations of electron density, their ultrastructural heterogeneity may be the expression of different states of secretory activity of one pinealocyte cell type. On the other hand, differences in the content of synaptic-like microvesicles in the process terminals of 'light' and 'dark' cells could also indicate a principal functional heterogeneity of the microvesicular compartment among pinealocytes, pointing to the existence of different types of pinealocytes. PMID- 8410080 TI - Early detection of post-traumatic olivary hypertrophy by MRI. AB - Two patients are described, in whom MRI detected unilateral signal abnormalities in the region of the inferior olivary nuclei, suggesting an early stage of olivary hypertrophy. MRI was performed 4 and 7 weeks respectively after traumatic brain-stem injury. Palatal myoclonus was concomitantly observed in one patient, while the other showed no evidence of segmental myoclonus at the time of examination. The authors conclude that MRI is highly sensitive in the detection of olivary hypertrophy and of traumatic lesions of the dentato-rubro-olivary pathway. PMID- 8410079 TI - Progressive multifocal leukoencephalopathy in AIDS: a clinicopathologic study and review of the literature. AB - We reviewed the clinical, radiographic, and pathologic features of 15 patients with the acquired immune deficiency syndrome (AIDS) and progressive multifocal leukoencephalopathy (PML). Brain tissue from 10 autopsy and 6 biopsy specimens was studied using: in situ hybridization (ISH) for JC virus (JCV), immunohistochemistry for human immunodeficiency virus (HIV) p24 antigen, and electron microscopy. Thirteen patients presented with focal neurologic deficits, while 2 presented with a rapid decline in mental status. PML was commonly the initial opportunistic infection of AIDS and produced hemiparesis, dementia, dysarthria, cerebellar abnormalities, and seizures. Magnetic resonance imaging was more sensitive than computed tomography in detecting lesions, and often showed multifocal areas of PML. CD4+ T-cell counts were uniformly low (mean 84/mm3), except in 1 patient who improved on 3'-azido-3'-deoxythymidine (AZT). PML involved the cerebral hemispheres, brain stem, cerebellum, and cervical spinal cord. The distribution of brain involvement was consistent with hematogenous dissemination of the virus. In 2 brain specimens, multiple HIV-type giant cells were present within the regions involved by PML. When co-infection by HIV and papovavirus was present, PML dominated the pathological picture. ISH for JCV showed virus in the nuclei of oligodendrocytes and astrocytes. Occasionally there was staining for JCV in the cytoplasm of glial cells and in the neuropil, the latter possibly a correlate of papovavirus spread between myelin sheaths, as seen by electron microscopy. ISH demonstrated more extensive foci of PML than did routine light microscopy. PMID- 8410081 TI - Melatonin, cortisol and body temperature rhythms in Lennox-Gastaut patients with or without circadian rhythm sleep disorders. AB - The daily rhythms of melatonin, cortisol and body temperature were studied in 16 institutionalized subjects with the Lennox-Gastaut syndrome. The results of 9 subjects with normal daily rhythms of sleep and wakefulness (group 1) were compared with those of 7 subjects with disordered sleep (group 2). Salivary samples were collected and axillary temperature was measured every 2 h during two or three separate 26-h periods. The hormones were measured by radioimmunoassays. The rhythms were characterized with single cosinor analysis. Two subjects in group 1 and six subjects in group 2 had abnormalities in their rhythms of temperature, cortisol or melatonin. All three rhythms were disrupted in two subjects of group 2. These two subjects were the only ones with disrupted cortisol rhythm. The diversity of rhythm pathologies suggested partly separate regulatory mechanisms for each rhythm. The co-occurrence of circadian rhythm sleep disorders with the deteriorated melatonin rhythm raised the question as to whether the sleep disorders of these subjects, like those of subjects with healthy brains, could be relieved by the induction of normal melatonin rhythm. PMID- 8410082 TI - Viral infections trigger multiple sclerosis relapses: a prospective seroepidemiological study. AB - A neurological surveillance was combined with prospective recording of upper respiratory and gastrointestinal infections and serological diagnosis of five common viral infections in 60 benign multiple sclerosis patients, with a mean follow-up of 31 months. During 4-week at risk (AR) periods encompassing common infections, a significant excess of MS relapses was found in the AR period, with a relative risk of 1.3. A seasonal variation of the MS relapse rate was found with a minimum in summer. There was a significant correlation between the number of AR relapses and the number of common infections per month explaining the periannual distribution of relapses. The non-AR relapses showed no seasonal variation. There was a significant correlation between adenovirus CF titre rises associated with upper respiratory infections and the occurrence of a major MS relapse in the AR period (n = 7), while influenza infections were not followed by a major MS relapse (n = 6). Linear homologies have been demonstrated between adenovirus and basic myelin protein. The epidemiological approach is essential to our understanding of systemic antigens triggering multiple sclerosis activity. PMID- 8410083 TI - Early MRI findings in Creutzfeldt-Jakob disease. AB - We describe the MRI changes preceding the onset of myoclonus in two patients whose post-mortem examination confirmed the diagnosis of Creutzfeldt-Jakob disease (CJD). MRI showed changes in the striatum early in the course of CJD (2-6 months after the onset of apathy, interpreted as depression, and 1-2 months before the onset of further clinical symptoms). Only in one patient did electroencephalography record the typical triphasic sharp-waves, 1 month after MRI. PMID- 8410084 TI - The utility of F wave chronodispersion in lumbosacral radiculopathy. AB - The sensitivity of F wave chronodispersion (Fc) in evaluating nerve root pathology is unknown. We compared Fc in 91 patients with clinical and EMG evidence of L5 or S1 radiculopathy with Fc in 81 controls in order to evaluate its sensitivity in lumbosacral radiculopathy. F waves were obtained by stimulating the peroneal and tibial nerves behind the knee and recording from the extensor digitorum brevis (L5 predominant) and flexor hallucis brevis (S1 predominant) muscles, respectively. Fc was calculated by subtracting the shortest F wave latency from the longest and, in controls, ranged from 0.2 to 23.4 ms in the peroneal nerve, and from 1.2 to 13.4 ms in the tibial nerve (95th percentile = 13 ms for the peroneal nerve and 9.2 ms for the tibial nerve). In the patient group, Fc also ranged from 0.2 to 23.4 ms in the peroneal nerve, and from 0.4 to 18.2 ms in the tibial nerve. Only 5 (5.5%) and 8 (11.3%) patients for the peroneal and tibial nerves, respectively, had Fc values which fell beyond the 95th percentile, a percentage far below the sensitivity of F wave latency measurement and not substantially different from chance. Thus we conclude that Fc has no substantial additional value in evaluating lumbosacral radiculopathy over that of F wave latency. PMID- 8410085 TI - A positron emission tomography study of patients with acute carbon monoxide poisoning treated by hyperbaric oxygen. AB - Seven patients with an acute and severe carbon monoxide intoxication were treated with hyperbaric oxygen and underwent a positron emission tomographic examination 2-5 days after the acute event. Although the final clinical outcome was good in all patients, ischaemic changes were observed. Three patients with temporary sequelae after hyperbaric oxygen treatment showed the most severe changes, mainly in striatum and thalamus. Although positron emission tomographic examination cannot predict the final outcome, it can show the regions at risk for development of late complications following carbon monoxide poisoning. PMID- 8410086 TI - Chronic fatigue: electromyographic and neuropathological evaluation. AB - Single fibre electromyography of extensor digitorum communis muscle (EDC) was performed on 35 patients with chronic fatigue, the majority of whom also had creatine kinase estimation and biopsy of EDC. The subjects were categorised as having an acute-onset post-viral fatigue syndrome, a non-specific chronic fatigue or possible muscle disease in view of pronounced myalgia. Of 11 subjects who had myalgia as a significant symptom, abnormalities in fibre density were found in 6, and 5 of these had some non-specific abnormalities on muscle biopsy, with creatine kinase levels being normal in all cases. Fibre density estimation may be a useful way of identifying a subgroup of chronic fatigue sufferers with a possible primary muscle disorder. PMID- 8410087 TI - Electrophysiological motor testing, MRI findings and clinical course in AIDS patients with dementia. AB - Thirty-three HIV-positive patients with clinical signs of dementia according to the 1991 AAN criteria underwent psychometric, electrophysiological and radiological examination and were compared with a group of normal healthy subjects and a cohort of clinically asymptomatic HIV-1-positive individuals of comparable education and social environment. Compared with the other groups, test performance was severely impaired in the demented patients. Results of motor testing and MRI revealed that subcortical structures were not exclusively affected, but most severely and early, thus characterizing the clinical feature in HIV-1-associated dementia. In demented patients a rapid deterioration was observed, leading to death within about 12 months on average, which is a markedly shorter survival time than described in the literature for non-demented HIV-1 positive individuals. PMID- 8410088 TI - Fatigue, sleep disturbances and circadian rhythm in multiple sclerosis. AB - The aim of this study was to investigate whether fatigue and sleep disturbances in multiple sclerosis (MS) patients might be due to disrupted circadian sleep wake regulation. Actigraphy and a multiple sleep latency test (MSLT) were performed in 16 MS patients with both prominent sleep complaints and fatigue. Actigraphy scores did not differ from control values, whereas sleep onset latency values were altered in subgroups of MS patients. No evidence was found for a generalized circadian disturbance in MS patients. PMID- 8410089 TI - Anti-myeloperoxidase antibodies in Churg-Strauss syndrome. PMID- 8410090 TI - Devic's neuromyelitis optica and varicella. PMID- 8410091 TI - An association between multiple sclerosis and diabetes mellitus. PMID- 8410092 TI - Adapting a clinical comorbidity index for use with ICD-9-CM administrative data: differing perspectives. PMID- 8410093 TI - The more things change.... PMID- 8410094 TI - Change in self-reported functioning in older persons entering a residential care facility. AB - We evaluated the responsiveness of measures of function in admissions to a long term care facility. Between baseline and follow-up assessment, one-fifth or more of the subjects either worsened or improved in most aspects of reported function. We compared two measures of self-reported function (COOP charts and a short-form survey). Convergent validity was observed for changes in pain, social health, and mental health (r = 0.39-0.74), but not for physical functioning. Although the short-form physical function measure discriminated worsening on several performance-based external criteria of physical functioning (area under ROC curves up to 0.82), the COOP and other measures of physical functioning were less likely to do so. All physical function measures were less responsive for detecting improvement. Clinicians and investigators intending to monitor change in function must consider the responsiveness of their measures. PMID- 8410095 TI - An evaluation of an asthma quality of life questionnaire as a measure of change in adults with asthma. AB - In assessing the effectiveness of management strategies for patients with asthma, it is important to measure outcomes which are relevant to the concerns of patients. Quality of life is one such outcome which may not be adequately reflected in lung function measurements. We have developed an asthma quality of life questionnaire (the AQLQ) for this purpose. The aim of this study was to test the validity and responsiveness of the AQLQ as a measure of change. Forty four adults with asthma were assessed on two occasions 4 months apart. On each occasion subjects completed the AQLQ and the Sickness Impact Profile (SIP). Lung function and the degree of bronchial hyperresponsiveness (BHR) were measured and diary cards were used to derive a symptom score and mean daily peak flow variability. The relation of change in AQLQ scores to change in the other outcomes was assessed. Questionnaire responsiveness was assessed by comparing the change in AQLQ scores between 19 improved and 20 stable subjects. Improvement was assessed on lung function and BHR criteria. As expected, change in AQLQ score was correlated with change in symptom score (r = 0.37, 95% CI -0.04 to 0.64) and change in BHR (r = 0.38, 95% CI 0.06 to 0.64). The associations with change in peak flow variability (r = 0.12, 95% CI -0.26 to 0.47) and change in SIP score (r = 0.18, 95% CI -0.12 to 0.45) were in the expected direction but weaker than expected. The AQLQ was capable of detecting differences between improved and stable subjects (p = 0.007).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410096 TI - Use of goal attainment scaling in measuring clinically important change in the frail elderly. AB - The selection of appropriate outcome measures is important in evaluating specialized geriatric programs, but how the various measures compare, and which are most appropriate, are matters still largely unexplored. We compared several outcome measures, including goal attainment scaling, to assess their sensitivity to changes in the health status of frail elderly patients admitted to two geriatric medicine wards. GAS is a measurement approach which accommodates multiple individual patient goals, and has a scoring system which allows for comparisons between patients. Forty-five patients (mean age 81 years, 30 females) received comprehensive assessments. GAS yielded a mean 5 goals per patient. The mean gain in the GAS score was 22 points (SD = 7) which was compared with the change in the Barthel Index (r = 0.59), the Functional Independence Measure (r = 0.45), the Physical Self-Maintenance Scale (r = 0.54), the Katz Activities of Daily Living Index (r = 0.49) and the Spitzer Quality of Life Index (r = 0.38). The inter-rater reliability of scoring the GAS follow-up guides was 0.91. Using a relative efficiency statistic, GAS proved more efficient than any other measure. The effect size statistic also demonstrated an increased responsiveness to change of GAS compared with standard measures. GAS is an individualized measurement approach which shows promise as a responsive measure in frail elderly patients. PMID- 8410097 TI - Moderate to high intensity conditioning leisure time physical activity and high cardiorespiratory fitness are associated with reduced plasma fibrinogen in eastern Finnish men. AB - A reduction of plasma fibrinogen has been suggested as one mechanism through which physical activity would protect against coronary heart disease (CHD). Therefore, we investigated the association of conditioning leisure time physical activity (CLTPA), assessed quantitatively by a 12-month history, and maximal oxygen uptake (VO2max) with plasma fibrinogen concentration in eastern Finnish men aged 42-60 years. A high mean intensity of CLTPA (standardized multivariate regression coefficient beta -0.059, p = 0.020) and a high maximal oxygen uptake (beta -0.163, p < 0.001) were associated with reduced plasma fibrinogen when adjusting for the strongest covariates. The adjusted relative difference in plasma fibrinogen concentration was 6.6% between men with a mean CLTPA intensity of < 4 and > 8 METs and 9.1% between the extreme quartiles of VO2max (< 2.21 vs > 2.961/min). The association between the mean intensity of CLTPA (p = 0.030 for interaction) and VO2max (p = 0.033) and plasma fibrinogen was stronger for smokers than for non-smokers. These data indicate that a reduction of plasma fibrinogen concentration may be one mechanism through which moderate to high intensity CLTPA and high cardiorespiratory fitness reduce the risk of CHD. PMID- 8410098 TI - High, usual and impaired functioning in community-dwelling older men and women: findings from the MacArthur Foundation Research Network on Successful Aging. AB - The objective of this study is to determine the range of complex physical and cognitive abilities of older men and women functioning at high, medium and impaired ranges and to determine the psychosocial and physiological conditions that discriminate those in the high functioning group from those functioning at middle or impaired ranges. The subjects for this study were drawn from men and women aged 70-79 from 3 Established Populations for the Epidemiologic Study of the Elderly (EPESE) programs in East Boston MA, New Haven CT, and Durham County NC screened on the basis of criteria of physical and cognitive function. In 1988, 4030 men and women were screened as part of their annual EPESE interview. 1192 men and women met criteria for "high functioning". Age and sex-matched subjects were selected to represent the medium (n = 80) and low (n = 82) functioning groups. Physical and cognitive functioning was assessed from performance-based examinations and self-reported abilities. Physical function measures focused on balance, gait, and upper body strength. Cognitive exams assessed memory, language, abstraction, and praxis. Significant differences for every performance based examination of physical and cognitive function were observed across functioning groups. Low functioning subjects were almost 3 times as likely to have an income of < or = $5000 compared to the high functioning group. They were less likely to have completed high school. High functioning subjects smoked cigarettes less and exercised more than others. They had higher levels of DHEA-S and peak expiratory flow rate. High functioning elders were more likely to engage in volunteer activities and score higher on scales of self-efficacy, mastery and report fewer psychiatric symptoms. PMID- 8410099 TI - Do communities differ in health behaviors? AB - Communities differ in the prevalence of various health behaviors, but it is not known to what extent these differences are due to "different types" of people living in them. We used data from the evaluation of the Henry J Kaiser Family Foundation Community Health Promotion Grant Program to study individual-level and community-level variation in health behaviors for 15 communities. Our results show (1) there was significant variation among these communities in prevalences of smoking, consumption of alcohol and dietary fat, and use of seatbelts; (2) these differences persisted after control for demographic, health status, and other health behavioral characteristics of the people in the communities; (3) the community effect on a particular person's behavior, as represented by R2, was very small (less than 1%); and (4) the adjusted differences in prevalences among communities were potentially large (for example, a 7 percentage point difference in the probability of smoking). Unique features of communities may influence health behaviors. These findings affirm the potential importance of contextual effects on individual health behavior and thus support the theory that changing the community environment may offer effective ways to change individual health behavior. PMID- 8410100 TI - Time-dependent variability in repeated measurements of cholesterol levels: clinical implications for risk misclassification and intervention monitoring. AB - Intraindividual variability (IIV) in total cholesterol levels based on measurements taken 1 week apart is compared with an estimate based on measurements taken 2 years apart. Single-subject 95% confidence intervals around the mean of two repeated measurements were Xi +/- 21 and +/- 28 mg/dl, respectively, and Xi +/- 30 and +/- 40 mg/dl for a single measurement. Comparing these results with published estimates over varying time intervals shows a trend of decreasing IIV with shorter intervals, suggesting that confidence interval widths based on short-term repeated measurements and those based on longer-term repeated measurements may differ more than previously assumed. The practical consequences are that: (1) the level of misclassification inherent in the National Cholesterol Education Program (NCEP) guidelines may be less than had been estimated; and (2) reliable cholesterol reductions resulting from dietary or other interventions may be somewhat easier to detect. These findings have implications for the cost-effectiveness of cholesterol screening strategies and interventions to reduce cholesterol. PMID- 8410101 TI - Myocardial infarction in Girona, Spain: attack rate, mortality rate and 28-day case fatality in 1988. Regicor Study Group. AB - This study was conducted to establish the attack rate, mortality rate and 28-day case-fatality rates of acute myocardial infarction (AMI) occurring in Girona, Spain, between October 1987 and September 1988. The study was carried out using a population register of AMI, and took place in one central hospital, and eight county hospitals in Girona (in the autonomous community of Catalonia, northeast Spain). Subjects comprised 222 fatal cases selected from 4069 death certificates, and 210 non-fatal cases from hospital records, among subjects aged between 25 and 74 years (reference population 263,778). The age standardized attack rate was 105.6 per 100,000 inhabitants in the age group 35-64 (193.6 in men and 17.6 in women). The age standardized mortality rate was 36.3 per 100,000 inhabitants in the age group 35-64 (63.1 among men and 9.9 among women). The age standardized 28 day case-fatality was 34.6% in men and 50.1% in women in this same age group. Attack and mortality rates of AMI in a region of Spain are presented for the first time. These rates are lower than in other developed countries, nevertheless the 28-day case-fatality is similar to that of these countries. PMID- 8410102 TI - Interpreting a single antistreptolysin O test: a comparison of the "upper limit of normal" and likelihood ratio methods. AB - Single serologic tests may occasionally influence clinicians in making diagnoses. The antistreptolysin O (ASO) test is a frequently used tool for detecting recent Streptococcus pyogenes infection and is helpful in the diagnosis of diseases like rheumatic fever. Using data from a 1989 prospective study of 600 healthy male military recruits, in which 43% experienced S. pyogenes upper respiratory tract infection (2-dilution rise in ASO), this report compared two methods of interpreting a single ASO titer. Using the "upper limit of normal" (80 percentile) method, recruits with an ASO titer of greater than 400 showed evidence of recent S. pyogenes infection. This method had a sensitivity and specificity of only 65.9 and 81.9% respectively. In contrast to the "yes-no" dichotomy of the "upper limit of normal" method, the likelihood ratio method statistics were ASO value specific, more consistent with clinical judgment, and better emphasized the caution clinicians must use in interpreting a single ASO test. PMID- 8410103 TI - Terminal digit preference, random error, and bias in routine clinical measurement of blood pressure. AB - We examined the presence, magnitude, and consequences of systematic and random errors caused by terminal digit preference in the measurement of highest systolic blood pressure during prenatal visits in 28,841 non-referred pregnant women who delivered between 1 January 1982 and 31 March 1990. In the overall distribution of terminal digit readings, 78% were read to 0, 15% to even digits other than 0, 5% to 5, and only 2% to odd digits other than 5. This preference for 0's was consistent across the entire distribution of blood pressure and for a variety of maternal characteristics. The relative frequency of the cutoff value of 140 mmHg (i.e. the percentage of readings on 140 mmHg) within the range containing the value (i.e. 138-142 mmHg) was similar to the relative frequency of other multiples of 0. This was true whether the comparison was made in the overall study sample, or in a pre-selected low-risk subgroup or high-risk subgroup, indicating no systematic bias. On the other hand, a strong tendency to read blood pressure values to the nearest 0 had a marked effect on the classification of hypertension. Changing the definition of hypertension from > or = 140 mmHg to > 140 mmHg produced a reduction in prevalence of hypertension from 25.9 to 13.3% in the overall study sample, from 15.4 to 6.3% in the low-risk subgroup, and from 43.3 to 25.3% in the high-risk subgroup. Epidemiologic studies that compare prevalences of hypertension in different populations based on routine clinical measurement of blood pressure and a single cutoff point should assess the consequences of terminal digit preference in defining hypertension.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410104 TI - The effects of joint misclassification of exposure and disease on epidemiologic measures of association. AB - This paper addresses the effects of simultaneous misclassification of both exposure and disease on epidemiologic measures of association. If misclassification of a dichotomous exposure is independent of a dichotomous disease status and vice versa (non-differential misclassification), and misclassification of exposure is independent of misclassification of disease, then the bias is always toward the null. In practice, however, errors in exposure and disease ascertainment may often be correlated. In this case, the observed exposure-disease association may be strongly biased in any direction even with non-differential misclassification. As an important corollary, the assertion commonly made in the discussion of epidemiologic study results that the observed measures of association can only be biased toward the null due to presumedly non differential misclassification has to be viewed as inadequate unless the assertion that exposure and disease misclassification are independent is also justified. Inferences regarding the degree and direction of bias due to misclassification of exposure and disease should consider plausible degrees of correlation in classification errors in addition to the overall misclassification rates. Whenever possible, sensitivity analyses should be performed to provide a quantitative basis for such inferences. PMID- 8410105 TI - The design of prospective epidemiological studies: more subjects or better measurements? AB - Prospective epidemiological studies which seek to relate potential risk factors to the risk of disease are subject to appreciable biases which are often unrecognized. The inability to precisely measure subjects' true values of the risk factors under consideration tends to result in bias towards unity in the univariate relative risks associated with them--the more imprecisely a risk factor is measured, the greater the bias. When correlated risk factors are measured with different degrees of imprecision the adjusted relative risk associated with them can be biased towards or away from unity. When designing a new prospective study cost considerations usually limit the total number of subject-evaluations that are available. The usual design approach is to maximize the study size and evaluate each subject on one occasion only. An alternative approach involves recruitment of a smaller number of subjects so that each can be evaluated on more than one occasion, thus resulting in a more precise measure of subjects' risk factor values and hence less bias in the relative risk estimates. In this paper we use a simulation approach to show that under conditions that prevail for most major prospective epidemiological studies the latter approach is actually more likely to produce accurate relative risk estimates. This emphasizes the importance of bias due to exposure measurement imprecision and suggests that attempts to anticipate and control it be given at least as high a priority as that given to sample size assessment in the design of epidemiological studies. PMID- 8410106 TI - Baldness and coronary heart disease risk factors. AB - The present report focuses on the association between baldness pattern and coronary heart disease risk factors in 872 male factory workers from southern Italy participating in an epidemiological study. Participants were divided according to presence or absence of baldness and baldness pattern. Participants with fronto-occipital baldness (male-type baldness) (n = 280) characterized by hair loss centered over the vertex with an m-shaped frontal-temporal recession had, on the average, higher serum cholesterol and blood pressure compared to participants with no baldness (n = 321) and/or participants with just frontal baldness (n = 273). For serum cholesterol, a significant interaction was detected between age and fronto-occipital baldness (i.e. the association between fronto occipital baldness and elevated levels of serum cholesterol became weaker with age). No interaction was detectable between age and fronto-occipital baldness for blood pressure. The results of this cross-sectional study indicate that male-type pattern of baldness is associated with elevated CHD risk profile, and that this relation between age and serum cholesterol differs in younger compared to older men. PMID- 8410107 TI - New problems for nursing in Europe's professional and working conditions. PMID- 8410108 TI - Photodynamic therapy: shining light where it is needed. PMID- 8410109 TI - A prospective phase II study on photodynamic therapy with photofrin II for centrally located early-stage lung cancer. The Japan Lung Cancer Photodynamic Therapy Study Group. AB - PURPOSE: A phase II study was conducted between June 1989 and February 1992 to evaluate the activity and toxicity of photodynamic therapy (PDT) with photofrin II in centrally located early-stage lung cancer and to determine the complete response (CR) rate as the primary end point. PATIENTS AND METHODS: Patients had histologically proven lung cancer and endoscopically superficial thickening or small protrusions. All lesions were located in subsegmental or larger bronchi. All patients had a performance status (PS) of 0 to 2 and arterial oxygen pressure tension (PaO2) > or = 60 mm Hg. No lymph node or distant metastases were present. All patients received photofrin II (2 mg/kg) intravenously 48 hours before PDT. Tumor lesions were superficially photoradiated by an argon dye laser or an excimer dye laser. RESULTS: Of 54 patients with 64 carcinomas, 51 with 61 carcinomas were eligible for toxicity evaluation and 49 with 59 carcinomas were assessable for response. Of the 59 assessable carcinomas, 50 (84.8%; 95% confidence interval, 73.0% to 92.8%) showed a CR after initial PDT. The median duration of CR was 14.0+ months (range, 2.0+ to 32.4+). The multiple regression model indicates that estimated length of longitudinal tumor extent was the only independent prognostic factor for CR (P = .002). Five carcinomas that had a CR had a local recurrence at 6, 10, 12, 16, and 18 months after initial PDT, respectively. Toxicity assessment (World Health Organization [WHO] grade 2) showed transient elevation of ALT (1.9%), pulmonary toxicity (7.7%), and allergic reaction (7.7%), as well as sunburn (1.9%). CONCLUSION: PDT with photofrin II has an excellent effect on patients with centrally located early-stage lung cancer who have limited tumor invasion extending over a small area (< or = 1 cm). PMID- 8410110 TI - Multiple-drug weekly chemotherapy versus standard combination regimen in small cell lung cancer: a phase III randomized study conducted by the European Lung Cancer Working Party. AB - PURPOSE: A randomized trial was conducted in patients with small-cell lung cancer (SCLC) to determine if survival can be improved by a weekly chemotherapy regimen combining various drugs. PATIENTS AND METHODS: Two hundred twenty-three patients were randomized to receive either six courses of a multiple-drug combination (MDC) regimen (Adriamycin [ADR; doxorubicin; Farmitalia Carlo Erba, Milan, Italy] 25 mg/m2 intravenously [i.v.] on day 1; etoposide [VP16] 120 mg/m2 i.v. on day 1; cyclophosphamide [CPA] 500 mg/m2 i.v. on day 1; cisplatin 60 mg/m2 i.v. on day 8; vindesine [VDS] 3 mg/m2 i.v. on day 8; vincristine [VCR] 2 mg i.v. on day 15; methotrexate [MTX] 100 mg/m2 i.v. on day 15), or a standard chemotherapy (SC) regimen (ADR 50 mg/m2 i.v. on day 1; CPA 1 g/m2 i.v. on day 1; VP16 80 mg/m2 i.v. on days 1 to 3). RESULTS: In 98 MDC-treated and 101 SC-treated assessable patients, we observed 69% and 62% objective responses rates, respectively. There was no significant difference in survival, with median durations and 2-year overall survival rates of 49 and 43 weeks and 8.5% and 7.9%, respectively. There was a significant increase in response rate in favor of MDC patients with limited disease (84% v 62%). Toxicity was tolerable, although SC was more hematotoxic, with 76% (v 59%) experiencing leukopenia and 17% (v 7%) experiencing thrombocytopenia (grades III and IV). If the cumulative doses received were nearly equal to the scheduled cumulative doses in both arms, the total relative dose-intensity (RDI) was significantly higher in the SC arm. The difference was due to increased treatment delays in the MDC arm. CONCLUSION: Weekly MDC failed to improve survival rates in patients with SCLC. PMID- 8410111 TI - Chemotherapy for advanced non-small-cell lung cancer: how much benefit is enough? AB - PURPOSE: To estimate the impact of chemotherapy on survival of patients with advanced non-small-cell lung cancer (NSCLC). METHODS: Randomized controlled trials (RCTs) published in the English-language medical literature between 1970 and 1991 were identified through MEDLINE and the reference lists of relevant articles. Six RCTs that accounted for 635 patients and compared first-line chemotherapy with supportive care in advanced NSCLC and reported survival up to at least 1 year were identified. Cumulative proportions of survival at 3, 6, 9, and 12 months for chemotherapy and control groups were derived from survival curves. RESULTS: Within each study, the effect of chemotherapy was estimated with a pooled relative risk (RR) across the four 3-month periods. An overall estimate of the RR of death at 1 year (RRM-H) was then calculated and a survival curve for chemotherapy-treated patients was constructed applying the pooled estimate of the RR (RRW) for each 3-month period. Overall, chemotherapy was associated with a 24% (95% confidence interval [CI], 13% to 34%) reduction in the probability of death when compared with supportive care. However, the effect of treatment appeared to decrease significantly after the first 6 months from therapy inception and the mean potential gain in survival, as compared with supportive care, was approximately 6 weeks (95% CI, 1 to 10). CONCLUSION: Chemotherapy is effective in the treatment of advanced NSCLC, but its impact on the length of survival is limited. Future RCTs should still include an untreated control group and should measure quality of life in addition to survival. PMID- 8410112 TI - Expression of epidermal growth factor receptor and survival in upper aerodigestive tract cancer. AB - PURPOSE: To determine the expression of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinoma and to evaluate its prognostic value. MATERIALS AND METHODS: EGFR was determined in tumor biopsies obtained from 109 consecutive patients with head and neck cancer (100 men, nine women). Control biopsies were obtained from 94 patients in a symetric nontumoral area of the same anatomic site. EGFR was measured by a binding assay using human recombinant iodine 125-EGF. RESULTS: The presence of detectable EGFR levels was found in all explored tumors with highly marked differences between patients (median, 71 fmol/mg protein; range, 2 to 2,302). In 93 of 94 cases, EGFR levels were higher in tumor samples as compared with healthy control zones. There was no significant difference in EGFR expression according to the various anatomic sites explored or tumoral differentiation status. There was a significant difference of distribution for EGFR levels between stages I and II tumors and stages III and IV tumors. The tumor EGFR levels were not linked to the response to first-line chemotherapy by cisplatin (CDDP) and fluorouracil (5FU). Survival was assessable for 103 patients for overall survival and for 81 patients for recurrence. EGFR overexpression was associated with shorter relapse-free (P = .0125) and overall survival (P = .028) rates. By multivariate analysis, the only significant variable was EGFR for relapse-free survival and tumor staging for overall survival. The association of EGFR to tumor staging markedly improves the significance for overall survival predictability (P = .002). CONCLUSION: EGFR determination deserves particular attention in head and neck cancer, since it independently carries a strong prognostic value. PMID- 8410113 TI - The benefit of leucovorin-modulated fluorouracil as postoperative adjuvant therapy for primary colon cancer: results from National Surgical Adjuvant Breast and Bowel Project protocol C-03. AB - PURPOSE: This study was designed to evaluate the efficacy of leucovorin-modulated fluorouracil (5-FU) as adjuvant therapy for patients with Dukes' stage B and C colon cancer. PATIENTS AND METHODS: Data are presented from 1,081 patients with Dukes' stage B and C carcinoma of the colon entered into National Surgical Adjuvant Breast and Bowel Project (NSABP) protocol C-03 between August 1987 and April 1989. Patients were randomly assigned to receive either lomustine (MeCCNU), vincristine, and 5-FU (MOF), or leucovorin-modulated 5-FU (LV + 5-FU). The mean time on study was 47.6 months. RESULTS: Comparison between the two groups indicates a disease-free survival advantage for patients treated with LV + 5-FU (P = .0004). The 3-year disease-free survival rate for patients in this group was 73% (95% confidence interval, 69% to 77%), compared with 64% (95% confidence interval, 60% to 68%) for patients receiving MOF. The corresponding percentage of patients surviving was 84% for those randomized to receive LV + 5-FU and 77% for the MOF-treated cohort (P = .003). At 3 years of follow-up, patients treated with postoperative LV + 5-FU had a 30% reduction in the risk of developing a treatment failure and a 32% reduction in mortality risk compared with similar patients treated with MOF. CONCLUSION: Treatment with LV + 5-FU significantly prolongs disease-free survival and results in a significant benefit relative to overall survival. These findings, when considered together with results from a recent meta-analysis demonstrating a benefit from LV + 5-FU in advanced disease, provide evidence to support the concept of metabolic modulation of 5-FU. PMID- 8410114 TI - A randomized, double-blind trial of fluorouracil plus placebo versus fluorouracil plus oral leucovorin in patients with metastatic colorectal cancer. AB - PURPOSE: A prospectively randomized trial was performed to determine whether the combination of fluorouracil (FU) plus leucovorin (FU-LV) administered orally is more effective than equitoxic FU for patients with metastatic colorectal cancer. PATIENTS AND METHODS: A double-blind, placebo-controlled trial design was used to eliminate observer bias. An escalating FU dosing schedule was used to achieve equal toxicity. End points were response, time to treatment failure (TTF), and eight quality-of-life (QL) parameters. A crossover arm allowed FU-treated patients to receive FU-LV combination treatment after treatment failure. RESULTS: Response rate was 32% for FU-LV versus 23% for FU (P = .15). Median TTF was 22 versus 16 weeks (P = .27). Median survival time was 44 versus 54 weeks (P = .26). QL was the same for both treatments, except for days of hospitalization, which was greater for FU-LV (P < .001). Toxicities were similar to those previously reported for FU-LV and FU alone. CONCLUSION: Oral LV-FU produces the same efficacy and toxicity pattern as has been reported for intravenous LV-FU. When FU LV is compared with equitoxic doses of FU, there is no difference in patient outcome. These results suggest that patients with advanced disease should receive FU at doses adequate to produce toxicity. PMID- 8410115 TI - Comparison of the prognostic significance between the number of metastatic lymph nodes and nodal stage based on their location in patients with gastric cancer. AB - PURPOSE: To determine which is the better prognostic determinant in gastric cancer: number of positive metastatic lymph nodes or current nodal stage. PATIENTS AND METHODS: Seven hundred seventy-seven patients who underwent potentially curative resections for gastric cancer were divided into three groups according to the depth of invasion. The influence of the number of positive nodes on their survival rate was analyzed. A multivariate analysis by the Cox proportional hazards model was used to determine independent prognostic factors. RESULTS: A decreased survival rate was associated with an increased number of positive nodes in all of the subjects and in each of the three groups. Patients with one to three positive nodes had as good a prognosis as those without nodal involvement when each of the three groups was analyzed separately. Using a multivariate analysis in the patients with four or more positive nodes, we found that the number of positive nodes was the most important prognostic determinant (P < .0001), followed by the depth of invasion (P < .02), and that the nodal stage was not significantly prognostic. Further multivariate analysis in the patients with one to three positive nodes showed that nodal stage and number of positive nodes were not significantly prognostic. CONCLUSION: The number of metastatic nodes should be adopted for classification of nodal stage in gastric cancer. PMID- 8410116 TI - Tumor-cell DNA content predicts outcome in children and adolescents with clinical group III embryonal rhabdomyosarcoma. The Intergroup Rhabdomyosarcoma Study Committee of the Children's Cancer Group and the Pediatric Oncology Group. AB - PURPOSE: The prognostic value of tumor-cell DNA content (ploidy) was evaluated in children with unresectable, nonmetastic rhabdomyosarcoma of embryonal histology. PATIENTS AND METHODS: Flow-cytometric techniques were used to estimate the ploidy of tumor specimens from 34 patients with embryonal rhabdomyosarcoma who were enrolled in the intergroup rhabdomyosarcoma study III (IRS III) from 1985 to 1991. Tumors were classified as diploid or hyperdiploid (DNA content, 1.1 to 1.8 times that of normal diploid cells). The influence of ploidy on clinical outcome was assessed by the Kaplan-Meier technique and Cox regression analysis with stepwise selection. RESULTS: Twelve of the tumor specimens were diploid and 22 were hyperdiploid. The patient groups defined by diploid or hyperdiploid tumors had similar presenting characteristics (eg, age, tumor size, and anatomic site). Significantly more children with hyperdiploid tumors achieved a complete response than did children with diploid tumors (85% v 42%; P = .01). The probability of progression-free survival at 5 years (+/- SE) was 91% +/- 6% for the hyperdiploid group, compared with 17% +/- 11% for the diploid group (P < .001). Hyperdiploidy was also associated with a significantly higher overall survival rate at 5 years: 96% +/- 4% versus 50% +/- 14% (P = .004). Ploidy retained its prognostic significance after adjustment for tumor site in the Cox regression model. CONCLUSION: Tumor-cell ploidy strongly correlates with outcome in children with nonmetastic, unresectable embryonal rhabdomyosarcoma. The two biologically distinct groups identified by this measure would benefit from further refinements in risk-directed therapy. PMID- 8410117 TI - Corrected QT interval prolongation in anthracycline-treated survivors of childhood cancer. AB - PURPOSE: Comprehensive cardiac evaluations are currently recommended for all anthracycline-treated patients to detect subclinical cardiac failure. A screening test is needed that would easily and inexpensively identify patients who are at risk for late cardiac decompensation. METHODS: We routinely reviewed the ECG and echocardiogram (ECHO) results of 52 of 56 anthracycline-treated long-term survivors of childhood cancer who had received > or = 100 mg/m2 of ANTH (ANTH = 1 mg/m2 of doxorubicin), and who were not in clinical heart failure. Exercise testing was performed in eight patients with a corrected QT interval (QTc) of > or = 0.43. RESULTS: Zero of 15 patients (without chest radiation) who received less than 300 mg/m2 of ANTH versus six of 22 who received > or = 300 mg/m2 of ANTH had a QTc > or = 0.43 (P = .03). Three of 15 patients (with chest radiation) who received less than 300 mg/m2 of ANTH versus 12 of 22 who received > or = 300 mg/m2 of ANTH had a QTc > or = 0.43 (P = .03). For all patients (including those with chest radiotherapy), zero of 19 who received less than 300 mg/m2 of ANTH versus eight of 33 who received > or = 300 mg/m2 of ANTH had a QTc of > or = 0.45 (P = .025). Three of 19 who received less than 300 mg/m2 of ANTH versus 19 of 33 who received > or = 300 mg/m2 of ANTH had a QTc of > or = 0.43 (P = .003). One patient had decreased fractional shortening (FS) and QTc prolongation. Cardiac decompensation (with a FS of 24%) occurred with propranolol in a patient with previously normal FS but prolonged QTc. With exercise, the QTc became further prolonged in all four patients with a QTc of 0.44 to 0.46 and in two of four patients with a QTc of 0.43. CONCLUSION: Prolongation of the QTc, a measure of myocardial repolarization, may reflect injury to myocardial cells. QTc prolongation may be predictive of an increased risk of late cardiac decompensation. If the utility of the QTc measure is confirmed, screening for evidence of myocardial damage can be easily and inexpensively performed by oncologists and primary caretakers. PMID- 8410118 TI - Total-body irradiation and autologous bone marrow transplant in the treatment of high-risk Ewing's sarcoma and rhabdomyosarcoma. AB - PURPOSE: In an effort to improve outcome in patients with metastatic or high-risk localized Ewing's sarcoma family of tumors (ESF) and rhabdomyosarcoma (RMS), we explored the role of consolidation therapy with total-body irradiation (TBI) plus autologous bone marrow transplantation (ABMT). PATIENTS AND METHODS: Ninety-one patients were entered onto one of three consecutive protocols from 1981 to 1986. Induction therapy consisted of four or five cycles of vincristine, doxorubicin, and cyclophosphamide (VAdriaC); in the earlier series, patients received one or two cycles with dactinomycin instead of doxorubicin. Irradiation of the primary site was used for local control. Patients who attained a complete response (CR) to induction therapy were eligible for consolidation with 8 Gy TBI plus VAdriaC and ABMT. RESULTS: Nineteen patients were ineligible for consolidation after failing to achieve or maintain a CR following induction therapy; all 19 are dead of disease. Seven eligible patients elected to forgo consolidation; three of seven are long-term event-free survivors. Sixty-five patients received consolidation therapy; 20 of 65 are long-term event-free survivors. A local control rate of 83% was achieved using radiation therapy as the primary modality of local control. Patients with metastatic disease at diagnosis fared substantially worse than did patients with localized tumors (6-year event-free survival [EFS] rate, 14% v 38%; two-sided P [P2] = .008). CONCLUSIONS: Consolidation of patients with metastatic or high-risk localized pediatric sarcomas with 8 Gy TBI plus ABMT has failed to improve the outcome of this group of patients. Metastatic disease at diagnosis continues to confer the poorest prognosis. New therapeutic strategies are needed to consolidate more effectively the remissions that can be achieved in the majority of these patients. PMID- 8410119 TI - Effectiveness of combined induction chemotherapy and radiotherapy in advanced nasopharyngeal carcinoma. AB - PURPOSE: This prospective trial was conducted with the goal of achieving an improvement in both overall and progression-free survival in previously untreated patients with stage IV nasopharyngeal carcinoma who received an induction chemotherapy regimen of fluorouracil (5-FU) and cisplatin followed by radiotherapy. PATIENTS AND METHODS: From January 1985 to January 1990, 47 patients with T1-4N2-3M0 squamous cell carcinoma of the nasopharynx were treated at The University of Texas M.D. Anderson Cancer Center with two to three cycles of 5-FU (1,000 mg/m2 continuous infusion per day x 5 days) plus cisplatin (100 mg/m2 continuous infusion on day 1 only) followed by radiotherapy using the conventional time/dose schedule. RESULTS: The response rate to chemotherapy was 93.2% (20.5% complete response [CR]; 72.7% partial response [PR]), and the overall CR rate after radiotherapy was 86%. With a median follow-up period of 53 months, the 2-, 4-, and 6-year survival rates were 80%, 71.6%, and 67.4%; the overall treatment failure rate was 27%. Treatment was well tolerated and without significant acute or chronic toxic effects. CONCLUSION: The results of this prospective study demonstrate that 5-FU plus cisplatin followed by radiotherapy can induce a durable remission in a high proportion of patients with poor prognosis stage IV nasopharyngeal carcinoma. PMID- 8410120 TI - Tumor necrosis is a prognostic predictor for early recurrence and death in lymph node-positive breast cancer: a 10-year follow-up study of 728 Eastern Cooperative Oncology Group patients. AB - PURPOSE: The Eastern Cooperative Oncology Group (ECOG) entered 766 patients onto two prospectively randomized surgical adjuvant clinical trials for lymph node positive breast cancer (T1-3N1M0). Ninety-five percent (n = 728) of eligible patients have complete information on the prognostic covariables under study (tumor necrosis [TN], tumor size, number of positive lymph nodes, age) and a median follow-up duration of 10.3 years. METHODS: TN was defined as confluent cell death in invasive areas of primary cancers, visible at 4 x objective lens magnification. Cox proportional hazards models were used to estimate presence versus absence of TN effects on clinical outcomes over full cross-stratification of variables, including delivery of chemotherapy versus observation only. Time varying effects were modeled using spline functions of time, and by fixing proportional hazards models separately in the time periods 0 to 2 and 2+ years. RESULTS: Presence of TN was an independent predictor for time to recurrence (TTR) (P = .007) and for survival (P = .0003) in the overall 10-year follow-up period. Presence of TN was also an independent predictor for TTR and for survival (each P < .0001) in the period 0 to 2 years after diagnosis. Spline function time modeling calculations showed different hazard ratios in the TN-present (TN+) versus TN-absent (TN-) groups for both TTR and survival (each P < .0001). This difference is changing over time (P = .0001 for TTR, P = .0005 for survival). Once a patient has been disease-free beyond 2 years after diagnosis, presence or absence of TN is irrelevant to future prognosis. CONCLUSION: Confluent TN of any dimension in invasive areas of lymph node-positive breast cancer is an independent predictor for early recurrence and death from the disease. PMID- 8410121 TI - Carboplatin plus ifosfamide as salvage treatment of epithelial ovarian cancer: a pilot study. AB - PURPOSE: This study aimed to evaluate the activity and toxicity of carboplatin (PPL) and ifosfamide (IFO) in patients with epithelial ovarian cancer previously treated with cisplatin (CDDP)-containing regimens. PATIENTS AND METHODS: From July 1989 to December 1991, 35 patients with epithelial ovarian cancer relapsed or refractory to CDDP as first-line chemotherapy were treated. PPL was administered at a dose of 300 mg/m2 intravenously (IV) on day 1 and IFO at a dose of 1,500 mg/m2 IV on days 1 to 3 every 3 to 4 weeks. Criteria for evaluating previous response to CDDP were strictly defined. RESULTS: The overall response rate was 43% (complete response [CR], 6%; partial response [PR], 37%) and the median duration of response was 7 months (range, 3 to 16). In potentially platinum-sensitive (PPS; relapsed) patients, the overall response rate was 56%. None of the primary platinum-resistant (PPR) patients obtained a clinical response to PPL plus IFO, whereas one of five secondary platinum-resistant (SPR) patients obtained a PR. The regimen was easily manageable. CONCLUSION: PPL plus IFO is useful and well-tolerated combination in salvage treatment of patients with advanced ovarian cancer. However, clear synergism between PPL and IFO that could overcome intrinsic or acquired CDDP resistance was not observed. The advantage of PPL plus IFO as compared with CDDP-containing regimens is represented by the increased tolerability and the reduced neurotoxicity, nephrotoxicity, and ototoxicity as compared with CDDP-containing regimens. It is essential that the patient population be defined according to their previous response to platinum therapy in trials involving second-line therapy of ovarian cancer. PMID- 8410122 TI - Randomized phase III trial of treatment with high-dose interleukin-2 either alone or in combination with interferon alfa-2a in patients with advanced melanoma. AB - PURPOSE: To compare the response rate, survival, and toxicity of treatment with high-dose intravenous (IV) bolus interleukin-2 (IL-2) plus interferon alfa-2a (IFN-alpha) with high-dose IL-2 alone in patients with advanced melanoma in a randomized phase III trial design. PATIENTS AND METHODS: Eighty-five patients with advanced melanoma were randomly assigned to receive IL-2 6 X 10(6) U/m2 per dose every 8 hours as tolerated for a maximum of 14 doses on days 1 through 5 and 15 through 19, or IL-2 4.5 X 10(6) U/m2 per dose, plus IFN-alpha 3 X 10(6) U/m2 using an identical schedule. A planned interim analysis was performed after 85 patients were entered, which forms the basis for this report. RESULTS: Partial response (PR) occurred in two of 44 patients (5%; 95% confidence interval, 1% to 15%) receiving IL-2 alone, compared with four of 41 patients (10%; 95% confidence interval, 3% to 23%) receiving IL-2/IFN-alpha (P = .30). There were no complete responses (CRs). The median duration of response was 11.5 months (range, 2.0 to 15.7+). There was no significant difference in the median survival duration for patients receiving IL-2 alone (10.2 months) compared with patients receiving IL 2/IFN-alpha (9.7 months). The median and mean number of doses of IL-2 were equivalent in both groups, as was toxicity. There were three treatment-related deaths, two in the IL-2-alone arm and one in the IL-2/IFN-alpha arm. The trial was terminated after the first interim analysis based on predefined early stopping rules, which included termination if the response rate in the IL-2/IFN alpha arm was less than 25%. CONCLUSION: Using the preparation, dose, and schedule of IL-2 in our trial, IFN-alpha failed to enhance significantly the response rate to high-dose IL-2 in the treatment of patients with advanced melanoma. PMID- 8410123 TI - Richter's syndrome: a report on 39 patients. AB - PURPOSE: The incidence, clinical features, laboratory findings, and treatment results of 39 patients with Richter's syndrome (RS) are reported. PATIENTS AND METHODS: Thirty-nine of 1,374 patients with chronic lymphocytic leukemia (CLL) developed RS. RESULTS: Features associated with RS included systemic symptoms (59%), progressive lymphadenopathy (64%), extranodal involvement (41%), elevation of lactate dehydrogenase (LDH; 82%), and a monoclonal gammopathy (44%). Analysis of the CLL karyotype showed no specific chromosomal abnormality that conferred increased risk; however, multiple abnormalities were common. Patients at all Rai stages and in complete response (CR) were at risk, including three CR patients with no residual disease at the level of detection by dual-parameter flow cytometry or restriction analysis for immunoglobulin (Ig) gene rearrangements. The incidence was not higher in patients who had received prior fludarabine or chlorodeoxyadenosine. The median survival duration was only 5 months, despite multiagent therapy. Patients who responded had prolonged survival durations (P < .001). Three of eight patients who survived more than 1 year had a de novo presentation of both CLL and large-cell lymphoma (LCL). Comparison of surface light-chain analysis from both low- and high-grade components demonstrated isotypic light-chain expression in 12 of 15 patients. Ig heavy- and light-chain gene rearrangement analysis showed identical rearrangement patterns in five of five patients. CONCLUSION: The clinical, laboratory, and survival characteristics of our RS patients were similar to those reported in earlier studies. Ig gene rearrangement and light-chain isotype analysis support a common origin for CLL and LCL. Despite progress in the treatment of CLL, the development of LCL remains a serious complication and continued surveillance in all CLL patients is warranted. PMID- 8410124 TI - Adult acute lymphoblastic leukemia: a multicentric randomized trial testing bone marrow transplantation as postremission therapy. The French Group on Therapy for Adult Acute Lymphoblastic Leukemia. AB - PURPOSE: In a prospective multicenter study, we analyzed the benefits of allogeneic bone marrow transplantation (BMT) in a nonselected group of adult patients with acute lymphoblastic leukemia (ALL) and, by a randomized trial, evaluated the effectiveness of autologous BMT over chemotherapy as postremission therapy in patients younger than 50 years who were not candidates for allogeneic BMT. PATIENTS AND METHODS: After induction therapy that randomized patients to receive one of two anthracycline-containing regimens, either daunorubicin (DNR) or zorubicin (ZRB), patients were assigned to postremission treatment according to age and results of HLA typing. Patients younger than 40 years with an HLA identical sibling (group 1) were scheduled to receive cyclophosphamide 60 mg/kg on days 1 and 2, total-body irradiation (TBI), and allogeneic BMT. Patients older than 50 years (group 2) received the chemotherapy arm composed of three monthly consolidation courses (DNR or ZRB, cytarabine, and asparaginase) followed by maintenance chemotherapy (modified L10 regimen). The remaining population (group 3) was randomly assigned to receive, after the three 1-month consolidation courses, either the chemotherapy arm or autologous BMT following a conditioning regimen similar to that of group 1. RESULTS: Of the 572 assessable patients, 436 achieved complete remission (78% +/- 2% for DNR v 74% +/- 3% for ZRB; P = .3). The estimated 3-year disease-free survival (DFS) rate for the 116 patients included in group 1 was 43% +/- 5%. Both autologous BMT (95 patients) and chemotherapy (96 patients) produced comparable 3-year DFS rates (39% +/- 5% v 32% +/- 5%) and survival durations (49% +/- 5% v 42% +/- 5%). However, late relapses after 36 months were mainly observed in the chemotherapy arm. CONCLUSION: This first interim analysis did not demonstrate a benefit of this autologous BMT procedure over classical maintenance chemotherapy in patients with ALL who received consolidation chemotherapy. PMID- 8410125 TI - Phase I clinical and pharmacokinetic evaluation of high-dose mitoxantrone in combination with cytarabine in patients with acute leukemia. AB - PURPOSE: To determine the maximally tolerated dose of mitoxantrone in combination with cytarabine in patients with acute leukemia and advanced phases of chronic myelogenous leukemia (CML), and to assess the pharmacokinetics of high-dose mitoxantrone in this patient population. PATIENTS AND METHODS: In a phase I study, 68 patients with acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), and accelerated- and blastic-phase CML received induction therapy consisting of cytarabine 3 g/m2 by infusion over 3 hours daily for 5 days, with escalating doses of mitoxantrone 40 to 80 mg/m2 over 1 to 2 days by intravenous infusion over 15 minutes. Mitoxantrone pharmacokinetics were evaluated by high performance liquid chromatography (HPLC) in 15 patients given a single dose of mitoxantrone ranging from 40 to 80 mg/m2 in combination with cytarabine. RESULTS: Severe, but reversible hyperbilirubinemia (> three times normal) was considered the dose-limiting toxicity, and was observed in 25% of all patients and in 35% of those who received 70 to 80 mg/m2 of mitoxantrone. Other extramedullary toxicity, including cardiac dysfunction, was mild. Myelosuppression was universal and the median time to complete remission (CR) was 28 days (range, 19 to 77). The CR rate for previously untreated and relapsed patients with AML was 85% (17 of 20) and 38% (seven of 18), respectively. Eighty-three percent (15 of 18) of patients with ALL achieved a CR, including all patients with previously untreated disease. Eight of 12 patients with advanced-phase CML achieved a CR. No significant changes in mean mitoxantrone plasma elimination rates (ie, terminal plasma half life and total-body clearance rate) occurred as the mitoxantrone dose doubled, indicating linear pharmacokinetics. CONCLUSIONS: The recommended phase II dose of mitoxantrone is 80 mg/m2 administered over 15 minutes as a single intravenous infusion in combination with cytarabine 3 g/m2/d for 5 days. At this dose, high concentrations of mitoxantrone are achievable in vivo to levels that have been shown to be extremely cytotoxic in vitro. PMID- 8410126 TI - Phase I dose-escalation trial of iodine 131-labeled monoclonal antibody OKB7 in patients with non-Hodgkin's lymphoma. AB - PURPOSE: Eighteen patients with recurrent or refractory CD21-positive, non Hodgkin's lymphoma (NHL) were treated in a phase IA dose-escalation therapeutic trial of iodine 131 labeled to a fixed dose of OKB7. METHODS: Individual doses of 30 to 50 mCi of 131I on 25 mg OKB7 were administered 2 to 3 days apart to achieve four total 131I-OKB7 dose levels of 90, 120, 160, and 200 mCi. Pharmacology, dosimetry, therapeutic effects, toxicity, human anti-mouse antibody (HAMA) response, and maximum-tolerated dose (MTD) were determined. Patients were evaluated by imaging studies (including whole-body gamma camera or single-photon emission computed tomography [SPECT] scans), flow cytometric analysis, bone marrow biopsy, and serial blood sampling. RESULTS: Median plasma and whole-body half-lives (T1/2) were 16 hours and 14 hours, respectively. Plasma and whole-body radiation doses were 0.0081 Gy/mCi and 0.0022 Gy/mCi, respectively. Specific tumor visualization was noted in eight of 18 patients. HAMA was detected in 12 of 16 patients. Nonhematologic toxicity was limited to asymptomatic elevations of thyroid-stimulating hormone (TSH) in five of 15 patients. Hematologic toxicity was observed in six of 18 patients, but was severe in only two patients. MTD in patients with diffuse lymphomatous bone marrow involvement was determined to be 200 mCi in four divided doses of 50 mCi 131I/25 mg OKB7. Antitumor activity was observed in 13 of 18 patients (one partial response [PR] and 12 mixed responses) and was dependent on the 131I-OKB7 dose administered. In general, palpable peripheral lymphadenopathy, enlarged spleens, skin lesions, and circulating OKB7 positive peripheral lymphocytes responded most readily to treatment. 131I-OKB7 was safely administered to a patient in leukemic phase of NHL with prompt subsequent loss of approximately 1 kg of tumor cells from the peripheral blood without associated tumor lysis syndrome. CONCLUSION: Because antitumor activity with tolerable toxicity was observed in the majority of this group of heavily pretreated patients, phase II investigation of mAb OKB7 radioconjugates in the therapy of NHL is warranted. PMID- 8410127 TI - Five-year administration of fenretinide: pharmacokinetics and effects on plasma retinol concentrations. AB - PURPOSE: Monitoring of fenretinide (4HPR) levels, kinetics, and effects on retinal was performed in patients who participated in a phase I trial and who continued to be treated for 5 years as phase III trial patients. Accumulation of 4HPR in the breast was also assessed. PATIENTS AND METHODS: Plasma concentrations of 4HPR, of its main metabolite N-(4-methoxyphenyl)retinamide (4MPR), and of retinol were assayed by high-performance liquid chromatography (HPLC) in breast cancer patients treated orally with 4HPR 200 mg/d for 5 years with a 3-day drug interruption at the end of each month. RESULTS: 4HPR, at 200 mg/d, resulted in average 4HPR plasma levels of approximately 1 mumol/L, which remained steady and caused steady retinol level reduction; 4MPR levels, similar to those of 4HPR, slightly but significantly increased during the first 35 months, but at 5 years they were similar to those at 5 months. During daily treatment, baseline retinol concentrations were reduced by 71%; after a 3-day drug interruption, all patients recovered and the mean reduction was 38%. After discontinuation of 5-year treatment, 4HPR and 4MPR half-lives (t1/2 beta) were 27 and 54 hours, respectively, similar to those reported after 28 daily treatments. After 6 and 12 months, the concentrations of 4HPR were at the limit of detectability (0.01 mumol/L), whereas those of 4MPR were five times higher. Baseline retinol concentrations were already recovered after 1 month. Accumulation of this retinoid in the breast was evidenced by concentrations of 4HPR and 4MPR in nipple discharge and in breast biopsies that were 10 and 20 times higher, respectively, than those found in plasma. CONCLUSION: 4HPR, at 200 mg/d for 5 years, resulted in constant drug plasma levels and constant retinol level reduction. After treatment interruption, 4HPR plasma concentrations decreased at the limit of detectability at 6 months and baseline retinol plasma concentrations were recovered after 1 month. PMID- 8410128 TI - Effect of dietary counseling on food intake, body weight, response rate, survival, and quality of life in cancer patients undergoing chemotherapy: a prospective, randomized study. AB - PURPOSE: This study examined the effect of frequent nutritional counseling on oral intake, body weight, response rate, survival, and quality of life in patients with cancer of the lung (small-cell), ovary, or breast undergoing cyclic chemotherapy. PATIENTS AND METHODS: Of 105 assessable patients, 57 were randomized to receive nutritional counseling, and 48 to receive no nutritional counseling and consumption of an ad lib oral intake. The intervention group was counseled to achieve a daily energy and protein intake according to recommended dietary allowances. Counseling was standardized and performed by a trained dietitian, and took place twice monthly during a 5-month period from start of chemotherapy. RESULTS: Dietary counseling increased daily energy intake by approximately 1 MJ and protein intake by 10 g over the entire study period. There was no change in the control group. Counseling led to an insignificant increase in body weight, but triceps skinfold measurement increased significantly after 5 months. Response rate and overall survival did not differ between the groups. Quality of life measured by the Quality-of-Life index (QL-index) increased significantly in both groups, but did not differ between groups. CONCLUSION: No clinical benefit could be demonstrated despite long-term and continuous improved food intake in cancer patients with solid tumors undergoing aggressive chemotherapy. PMID- 8410129 TI - Prognostic value of performance status and quality-of-life scores during chemotherapy for advanced breast cancer. The Australian New Zealand Breast Cancer Trials Group. PMID- 8410130 TI - Secondary cytoreduction: can it be justified? PMID- 8410131 TI - Postchemotherapy rheumatism: is this a menopausal symptom? PMID- 8410132 TI - More postchemotherapy rheumatism. PMID- 8410133 TI - Hepatic artery chemotherapy for colorectal liver metastases: yet more controversy. PMID- 8410134 TI - Splenectomy and risk of secondary leukemia. PMID- 8410135 TI - Ten years' experience with LSA2-L2 therapy for childhood advanced lymphoblastic lymphoma. PMID- 8410136 TI - Inhibition of cell proliferation and glutathione S-transferase by ascorbyl esters and interferon in mouse glioma. AB - Mouse glioma-26 (G-26) cell line established in this laboratory was used in the study. The in vitro effect of ascorbyl esters, viz., ascorbyl-palmitate (As-P), stearate (As-S) and mouse interferon-alpha/beta (MulFN-alpha/beta) on the glioma cell viability, proliferation and glutathione S-transferase (GST) activity was investigated. Cell viability and proliferation were examined by colorimetric MTT assay and [3H]-thymidine incorporation, respectively. Incubation (24h) of G-26 cells with As-S, As-P or MulFN-alpha/beta, resulted in a dose dependent decrease in cell viability (IC50 = 125 microM As-S; 175 microM As-P and 3.6 x 10(4) U/ml MulFN-alpha/beta) and proliferation (IC50 = 157 microM As-S; 185 microM As-P and 3.6 x 10(4) U/ml MulFN-alpha/beta). A combined exposure to 175 microM As-S and 800 U/ml of MulFN-alpha/beta resulted in a greater than an additive effect on cell viability and proliferation. The inhibition of cell proliferation/viability by interferon was species specific and was observed only with homologous MulFN alpha/beta, but not with human interferon-alpha lymphoblastoid or human interferon-beta. Ascorbyl esters inhibited cytosolic GST activity (1-50 = 15.0 microM As-S and 28.5 microM As-P) towards 1-chloro-2,4-dinitrobenzene in a dose dependent manner. The apparent Ki values for affinity purified GST, deduced from Dixon plots were 0.95 microM and 2.0 microM for As-S and As-P, respectively. Significant inhibition of GST was also observed in the cytosol isolated from G-26 cells exposed to 300 microM As-S or 800 U/ml MulFN-alpha/beta. PMID- 8410137 TI - Intracerebral and subcutaneous xenografts of human SCLC in the nude rat: comparison of monoclonal antibody localization and tumor infiltrating lymphocytes. AB - In the WAG/Rij nude rat, subcutaneous (s.c.) and intracerebral (i.c.) xenografts of the human SCLC cell line GLC-28 were evaluated for their growth behavior, in vivo monoclonal antibody binding and presence of tumor infiltrating lymphocytes. For the i.c. xenografts, two models of cerebral tumor growth were studied, one in the cerebral cortex and one in the lateral ventricle of the brain. In the s.c. and both i.c. xenografts models, in vivo localization of anti-carcinoma moab MOC 31 occurred within 4 hours after i.p. injection, with a maximal binding at 24 h after injection. A pronounced tumor infiltration of predominantly NK cells was observed for s.c. and intraventricular xenografts, but not for the GLC-28 tumors xenografted in the cerebral cortex. The presented nude rat/GLC-28 xenograft models may be used for the in vivo testing of experimental imaging techniques or alternative treatment strategies relevant to brain metastases of human SCLC. PMID- 8410138 TI - Immunocytochemical detection of calcineurin and microtubule-associated protein 2 in central neurocytoma. AB - An immunohistochemical study was carried out on four cases of central neurocytoma, which had characteristic clinicopathological features including ultrastructural findings. Specific antibodies to calcineurin (CaN), microtubule associated protein 2 (MAP2) and synaptophysin (SYP) were used. All tumor tissues examined showed specific immunoreactivity for CaN and MAP2. Immunolabelling of both molecules revealed that they were mainly localized in the perikarya and proximal processes of the tumor cells. SYP immunoreactivity was found in three of the four cases. SYP immunoreaction products were predominantly seen in the tumor cell processes, while the perikarya were weakly or moderately positive for SYP. The data suggest that CaN and MAP2, together with SYP, can be useful tools for identifying and characterizing of the central neurocytoma. PMID- 8410139 TI - Human glioblastoma cells produce 77 amino acid interleukin-8 (IL-8(77)). AB - The production of interleukin 8 (IL-8), a neutrophil chemotactic factor, and its amino acid sequence were examined in glioblastoma cell lines in vitro. Neutrophil chemotactic activity was demonstrated in 9 conditioned media of 15 human glioblastoma cell lines. Tumor necrosis factor (TNF)-alpha stimulated secretion of the activity in 7 lines and induced secretion in 4 other lines. ELISA quantification disclosed that the conditioned media contained interleukin 8 (IL 8) in an amount equivalent to the chemotactic activity. The IL-8 secretion increased with the stimulation by TNF-alpha. Northern blot analysis and the RT PCR method confirmed expression of mRNA in the glioblastoma cells and its augmentation by TNF-alpha and/or IL-beta. Reversed-phase HPLC following ion exchange chromatography revealed that the chemotactic activity was a single peptide, which was determined to be IL-8 by the retention time and ELISA. Furthermore, amino acid analysis disclosed that a major part of the glioblastoma cell derived IL-8 peptide was 77 amino acid IL-8 (IL-8(77); with the N-terminal sequence AVLPRSAKELRCQCI-). PMID- 8410140 TI - Chronic lymphocytic leukemia with CNS involvement. AB - Direct involvement of the brain by chronic lymphocytic leukemia (CLL) is extremely rare. This is the second documented case. A 63 year-old-man presented with motor deficits. Conventional radiographic imaging showed no abnormalities but an MRI scan of the head showed a high signal intensity lesion in the left cerebellum. Biopsy of this area showed a perivascular infiltrate of small B lymphocytes consistent with CLL involvement. The patient was treated with radiotherapy and had some improvement in symptoms. Three years later he remains alive with some neurologic deficit. This case confirms that CLL can be complicated by direct brain involvement. PMID- 8410141 TI - Pituitary oncocytomas: clinical features, characteristics in cell culture, and treatment recommendations. AB - To determine whether there are significant differences between oncocytomas and pituitary adenomas, we evaluated clinical features, treatment regimens and outcome in 23 males and 9 females (average age 64 years, range 43-81 years) with the histologic diagnosis of pure pituitary oncocytomas (> 95% oncocytes). Symptom duration was six to twelve months in 6 cases (19%) and more than one year in 19 cases (59%). Three patients presented with sudden onset of symptoms, and were found to have hemorrhage within their tumors. Visual loss (69%) and symptoms of hypopituitarism (44%) were the most common presenting complaints. Preoperative endocrine profiles revealed abnormalities in most cases, including pituitary insufficiency in 56% and hyperprolactinemia in 59%. The tumors were typically large at presentation; all but one had suprasellar extension. 28 patients underwent transsphenoidal tumor resections; 4 underwent subfrontal craniotomies. Gross dural invasion was found at surgery in 11 cases. At a mean followup of 31 months (range 2-68 months), recurrent tumor was identified in 4 patients (12.5%). Tumor size, dural invasion, preoperative endocrine profile, and postoperative radiotherapy did not correlate with recurrence. Among seven oncocytomas grown in culture, five demonstrated two distinct cell types consisting of oncocytes and typical adenoma cells, respectively. Oncocytomas often have a different clinical presentation than functional pituitary adenomas. PMID- 8410142 TI - Central neurocytoma: report of 2 cases and literature review. AB - Two cases of central neurocytoma are presented. This is a rare CNS tumour affecting, predominantly, young individuals. There is enough evidence in the literature now to distinguish this entity clinically, radiologically and histologically. Much less is known about the management of patients with this tumour, because most of the reports are in pathological literature. Using our two cases and reviewing the available literature, we are endeavouring to shed light on the clinical aspects and management of central neurocytoma. In particular, we attempted to evaluate the role of surgical excision and radiation therapy in the management of this tumour. PMID- 8410143 TI - Intrathecal IgM response in disseminated cerebrospinal metastasis from malignant melanoma. AB - Neurological complications are a major cause of morbidity and mortality in patients with disseminated malignant melanoma. We have studied and correlated clinical and cerebrospinal fluid (CSF) findings in 20 patients with central nervous system metastases from malignant melanoma including 8 patients with metastatic meningeal melanomatosis (MMM) and 12 patients with solid cerebral metastases (SCM). The putative CSF tumor markers, fibronectin and beta 2 microglobulin, were elevated significantly in MMM but not in SCM patients. A prominent increase in the IgM index, which reflects intrathecal B-cell stimulation, and a rise of IgG index, interleukin-6, and tumor necrosis factor alpha in MMM patients provide preliminary evidence for a local intrathecal immune response triggered by melanoma cell invasion of the subarachnoid space. PMID- 8410144 TI - cis-diamminedichloroplatinum (CDDP) therapy for brain metastasis of lung cancer. I. Distribution within the central nervous system after intravenous and intracarotid infusion. AB - The distribution of cis-diamminedichloroplatinum (CDDP) was studied in 23 patients undergoing surgical resection of brain tumors metastatic from lung cancer. CDDP (100 mg/m2) was administered intravenously (i.v.) or intra arterially (IA) at the time of surgery, and various fluids and tissues were sampled for measurement of drug concentration. Comparison of the two routes of administration disclosed that the plasma level was slightly lower after IA than after i.v. infusion, whereas there was no difference between the two routes in terms of drug diffusion into the brain tissue adjacent to the tumor. However, IA administration resulted in an intratumoral drug concentration twice as high as that achieved with i.v. infusion. The tumor:plasma and tumor:adjacent brain ratios of drug concentration after IA injection were also twice those measured after i.v. administration. The distribution pattern of CDDP is characteristic of water-soluble agents. All patients experienced tolerable nausea and vomiting. Creatinine clearance was moderately reduced in ten cases, but no serious renal toxicity was observed. Seizures occurred postoperatively in nine patients. Infrequent side effects were myelosuppression, ototoxicity, and postoperative intracranial bleeding. All adverse effects disappeared with conservative treatment or no intervention. PMID- 8410145 TI - cis-diamminedichloroplatinum (CDDP) therapy for brain metastasis of lung cancer. II: Clinical effects. AB - Parenchymal brain tumors, which were metastases of primary lung cancer, were surgically removed from 25 patients. During the operation, patients were administered (intravenous or intracarotid) 100 mg/sq m of cis diamminedichloroplatinum (CDDP) and postoperatively, they received intravenous CDDP at 3-month intervals for 1 year. The results of this mode of treatment were compared with those obtained in 25 patients who underwent the same surgery but received other anticancer agents and in 39 patients who received no postoperative chemotherapy. Patients in the CDDP-treated group survived much longer than both of the other treatment groups. In the CDDP, but not in the other two groups, survival was significantly longer in patients who had undergone resection of their lung tumors than in those who had not. The stage of lung cancer was not found to significantly influence survival time among CDDP-treated patients. Brain metastasis was the cause of death in 12% of the patients who received CDDP, in 16% those treated with other drugs, and in 26% of those who received no chemotherapy. The incidence of local and remote intracranial tumor recurrence, including meningeal carcinomatosis, was similar in the three groups. However, the mean interval from resection of the metastatic brain tumor to local or remote recurrence was longer in the CDDP-treated group than in the other two groups, and the 2-year-survival rate was significantly higher after CDDP administration. These results suggest that CDDP may be useful in the therapy of metastatic brain tumors derived from lung cancer. PMID- 8410146 TI - Brain metastases of metastatic malignant melanoma: response to DTIC and interferon-gamma. AB - The prognosis of patients with malignant melanoma and brain metastases is poor. Therapy of brain metastases is difficult and mostly unsuccessful, with brain metastases being the predominant factor which determines overall survival. We report here on a patient whose brain metastases responded to DTIC + INF-gamma. We present a short summary on the different effects on INF-alpha and INF-gamma and reach the conclusion that clinical trials which combine DTIC and INF-gamma should be performed. Based on this observation, combinations including INF-alpha are not necessarily comparable to modalities which include INF-gamma. PMID- 8410147 TI - Relationship of perfusion to edema in the 9L gliosarcoma. AB - The relationship between tumor perfusion and edema was analyzed, with edema characterized as tumor wet/dry weight ratio. Perfusion of subcutaneous 9L gliosarcoma was measured by injection of 133Xe in saline into the tumor core, followed by gamma camera imaging of 133Xe washout kinetics. A significant inverse correlation was found between edema and tumor perfusion (p < 0.0002), suggesting that edema can limit tumor perfusion, perhaps through a mechanism of increased interstitial fluid pressure. The perfusion rate of highly edematous tumors was reduced to less than 10% of the perfusion rate of less edematous tumors (p < 0.001). It was also found that tumor edema increased significantly with increasing tumor volume (p < 0.001), which could account for the finding that perfusion declined significantly with increasing tumor volume (p < 0.02). These findings are potentially important because it is possible to quantify tumor edema in vivo, with millimeter resolution, using 1H magnetic resonance imaging (MRI). Thus MRI may provide a non-invasive technique for characterizing tumor perfusion or tumor drug delivery. PMID- 8410148 TI - Role of the human anterior cingulate cortex in the control of oculomotor, manual, and speech responses: a positron emission tomography study. AB - 1. Two experiments were aimed at investigating the functional organization of the human anterior cingulate cortex (ACC) in relation to higher-order motor control. 2. The 15O-labeled H2O bolus method was used to measure relative changes of regional cerebral blood flow (rCBF) in 18 healthy human subjects as they performed oculomotor, manual, or speech tasks. 3. Task-specific rCBF changes were obtained in distinct subregions of the ACC, depending on the output system employed. The oculomotor and the manual task-related foci were found in the rostral and caudal regions of the ACC, respectively, whereas the speech foci were localized within two cingulate subregions, the intermediate dorsal and the rostral ACC. 4. In the manual tasks, two groups of activation foci could be distinguished, one just behind and the other just in front of the vertical plane traversing the anterior commissure. 5. The above pattern of rCBF changes was observed only if there was concomitant activation within the lateral prefrontal cortex (except for the posterior group of foci obtained in the manual tasks). 6. The localization of output-specific rCBF changes within the human ACC is consistent with the known somatotopic organization of the cingulate cortex in the monkey. 7. It is tentatively proposed that the ACC participates in motor control by facilitating the execution of the appropriate responses and/or suppressing the execution of the inappropriate ones. Such a modulatory effect would be of particular importance when behavior has to be modified in new and challenging situations. PMID- 8410149 TI - Spatial patterns of increased spinal cord membrane-bound protein kinase C and their relation to increases in 14C-2-deoxyglucose metabolic activity in rats with painful peripheral mononeuropathy. AB - 1. Three-dimensional spatial patterns of changes in membrane-bound protein kinase C (PKC) were examined in the lumbar spinal cords (L1-L5) of rats with an experimental painful peripheral mononeuropathy. Painful peripheral mononeuropathy was produced by loosely ligating the rat's common sciatic nerve, resulting in chronic constrictive nerve injury (CCI). Changes in spinal cord membrane-bound PKC distribution were assayed by employing an established quantitative [3H] phorbol-12,13-dibutyrate ([3H]PDBu) autoradiographic assay, which includes spinal cord sectioning, incubation of spinal cord sections with [3H]PDBu, production of autoradiographs, and computer-assisted image processing. 2. Sciatic nerve ligation induced demonstrable thermal hyperalgesia in response to radiant heat stimulation and spontaneous pain-related behaviors (such as lifting of the nerve ligated hind paw) in CCI rats 3, 7, and 10 days after unilateral sciatic nerve ligation. 3. Consistent with behavioral changes, CCI rats examined 3 or 10 days after sciatic nerve ligation displayed a three-dimensional pattern of increased membrane-bound PKC in the lumbar spinal cord (L1-L5) strikingly different from that of sham-operated rats: in the dorsoventral dimension, reliable increases in membrane-bound PKC occurred mainly within spinal cord laminae I-IV and V-VI in CCI rats; in the ipsilateral-contralateral dimension, changes in membrane-bound PKC were seen on both sides of the spinal cord in CCI rats with reliably higher levels of membrane-bound PKC on the side ipsilateral than on the side contralateral to sciatic nerve ligation; in the rostrocaudal dimension, increases in membrane-bound PKC in the spinal cord dorsal horns of CCI rats extended from spinal segments L2-L5. 4. Both three-dimensional increases in spinal cord membrane-bound PKC and nociceptive behaviors (thermal hyperalgesia and spontaneous pain behaviors) in CCI rats were reliably reduced after three daily intrathecal treatments with 80 nmol GM1 ganglioside (a glycosphingolipid shown to prevent PKC translocation/activation), the first of which was given 1 h after sciatic nerve ligation. This reduction was seen 24 h but not 7 days after the last GM1 ganglioside treatment. 5. This three-dimensional increase in membrane bound PKC in the spinal cord dorsal horn of CCI rats displayed high correlations with thermal hyperalgesia and with spontaneous pain-related behaviors in CCI rats observed both 3 and 10 days after sciatic nerve ligation. Similar correlations were observed between decreases in levels of membrane-bound PKC in the spinal cord dorsal horn and the attenuation of nociceptive behaviors in CCI rats after three daily intrathecal treatments with GM1 ganglioside.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410150 TI - Representation of the urinary bladder in the lateral thalamus of the cat. AB - 1. In alpha-chloralose-anesthetized cats the region surrounding the ventral posterolateral nucleus (VPL) of the thalamus was investigated to locate foci with input from the urinary bladder stimulated by application of intravesical pressure. The locations of the recording sites were verified in Nissl-stained histological sections with reference to electrolytic lesions. 2. Of the 23 visceroceptive thalamic neurons identified, 4 (17%) were located in the periphery of the VPL (VPLp) and 19 (83%) in the lateral and dorsal aspects of the posterior complex (POl and POd, respectively) adjoining VPLp. 3. The neurons responded to noxious intensities of intravesical pressure in the range of 50-100 mmHg. Excitatory responses were elicited in 8 (35%) neurons, "inhibitory" responses in 13 (57%) neurons, and 2 (9%) neurons responded with an increase and a decrease of their discharge to subsequent stimuli. 4. Of the 22 visceroceptive thalamic neurons tested for this parameter, 73% had low-threshold cutaneous receptive fields (RFs). These were located in the region of the lower back, the hip, the thigh, and the proximal tail (12 PO neurons), or covered the entire postcranial contralateral part of the body (3 PO neurons). For only one of the VPLp neurons, a somatic RF was found and this was located on the distal tail. The neurons responded to tap stimuli applied at a low repetition rate. None of the 11 neurons tested with noxious pinching of the skin was activated by this kind of stimulus. 5. It is concluded that the cat's lateral thalamic region, around but not within VPL proper, contains neurons that play a role in the processing of information about noxious events in the urinary bladder. A comparison with results from experiments in the monkey indicates differences in the organization of the visceroceptive systems between both species, regarding the thalamic localization of visceroceptive neurons, the occurrence of convergent low-threshold somatic RFs, and the association of excitatory and inhibitory effects of urinary bladder stimulation with the location of somatic RFs. PMID- 8410151 TI - Effects of ear plugging on single-unit azimuth sensitivity in cat primary auditory cortex. I. Evidence for monaural directional cues. AB - 1. Single-unit recordings were carried out in primary auditory cortex (AI) of barbiturate-anesthetized cats. Neurons, sensitive to sound direction in the horizontal plane (azimuth), were identified by their responses to noise bursts, presented in the free field, that varied in azimuth and sound pressure level (SPL). SPLs typically varied between 0 and 80 dB and were presented at each azimuth that was tested. Each azimuth-sensitive neuron responded well to some SPLs at certain azimuths and did not respond well to any SPL at other azimuths. This report describes AI neurons that were sensitive to the azimuth of monaurally presented noise bursts. 2. Unilateral ear plugging was used to test each azimuth sensitive neuron's response to monaural stimulation. Ear plugs, produced by injecting a plastic ear mold compound into the concha and ear canal, attenuated sound reaching the tympanic membrane by 25-70 dB. Binaural interactions were inferred by comparing responses obtained under binaural (no plug) and monaural (ear plug) conditions. 3. Of the total sample of 131 azimuth-sensitive cells whose responses to ear plugging were studied, 27 were sensitive to the azimuth of monaurally presented noise bursts. We refer to these as monaural directional (MD) cells, and this report describes their properties. The remainder of the sample consisted of cells that either required binaural stimulation for azimuth sensitivity (63/131), because they were insensitive to azimuth under unilateral ear plug conditions or responded too unreliably to permit detailed conclusions regarding the effect of ear plugging (41/131). 4. Most (25/27) MD cells received either monaural input (MD-E0) or binaural excitatory/inhibitory input (MD-EI), as inferred from ear plugging. Two MD cells showed other characteristics. The contralateral ear was excitatory for 25/27 MD cells. 5. MD-E0 cells (22%, 6/27) were monaural. They were unaffected by unilateral ear plugging, showing that they received excitatory input from one ear, and that stimulation of the other ear was without apparent effect. On the other hand, some monaural cells in AI were insensitive to the azimuth of noise bursts, showing that sensitivity to monaural directional cues is not a property of all monaural cells in AI. 6. MD-EI cells (70%, 19/27) exhibited an increase in responsiveness on the side of the plugged ear, showing that they received excitatory drive from one ear and inhibitory drive from the other. MD-EI cells remained azimuth sensitive with the inhibitory ear plugged, showing that they were sensitive to monaural directional cues at the excitatory ear.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410152 TI - Contribution of edema to the sensitization of high-threshold mechanoreceptors of the goat palatal mucosa. AB - 1. Carrageenan has been shown to alter transduction capacities of high-threshold mechanoreceptors of the goat palatal mucosa. Assessments were made of the contributions of carrageenan-induced edema to the sensitization of mechanonociceptors (mechanically insensitive afferents) and intense pressure receptors of the palatal mucosa of the goat. The degree of edema was assessed by measurement of changes in tissue compliance produced by carrageenan. The impact of a simulated edema on both dynamic and static aspects of transduction were examined. The following observations were made. 2. Tissue compliance of the ventral zone of the incisal papilla (IP) was decreased after carrageenan treatment (0.042-0.014 mm/g). Tissue compliance changes were large for small displacements (< 1.4 mm), where displacement curves were substantially linear. For large displacements (> 1.4 mm), there was an exponential relationship between displacement and reactive force, and the compliance was unchanged. The exponential component may have represented the contribution of the maxillary bone compliance measures. 3. A simulated edema was created to assess the effects of edema on afferent reactivity. Saline was used to create the simulated edema by using changes in tissue compliance as a means of matching simulated to carrageenan-induced edema. An analysis of linear and exponential phases of saline or carrageenan-induced changes in tissue compliance indicated that saline injections produced a good approximation of the changes in linear properties, but diverged from a match of the exponential phase of carrageenan edema. 4. Assessments were made of modifications in nociceptor reactivity after simulated edema or retested controls (n = 47). There was little indication that edema influenced quantitative aspects of either static or dynamic force transduction. Mechanonociceptors, but not intense pressure receptors, sensitized after repeated intense stimulation (mean decline of dynamic activation threshold of 42 +/- 16 g/mm2, mean +/- SE). Artificial edema did not further contribute to this sensitization. 5. Qualitative features of stimulus transduction were modified by simulated edema. Eleven of 17 afferents without initial static or dynamic transduction capacity acquired the capacity after artificial edema. 6. Although most aspects of force transduction were unaffected by artificial edema, displacement transduction was considerably shifted. Artificial edema decreased the threshold and shifted the displacement range over which nociceptors transduced dynamic and static aspects of force (threshold decreased from 1,894 +/ 229 to 1,456 +/- 194 microns; range decreased 239 +/- 99 microns). 7. It was concluded the contribution of edema to allodynia and hyperalgesia is best understood in terms of its contribution to population-coding mechanisms that are brought about by changes in transmission properties.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410153 TI - Abnormalities of mechanoreceptors in a rat model of neuropathic pain: possible involvement in mediating mechanical allodynia. AB - 1. Peripheral nerve injury sometimes leads to the development of neuropathic pain. One of the symptoms of such neuropathic pain is mechanical allodynia, pain in response to normally innocuous mechanical stimuli. We hypothesized that sympathetically driven dysfunction of cutaneous mechanoreceptors is responsible for signaling mechanical allodynia. The present study was undertaken to identify the types of sensory receptors that potentially mediate mechanical allodynia, with the use of a rat neuropathic pain model we have developed. 2. One week to 10 days after tight ligations of the L5 and L6 spinal nerves on one side, the rats fully developed behavioral signs of mechanical allodynia on the affected hindlimb. Various cutaneous mechanoreceptors originating from the neuropathic foot were examined by single-fiber recordings from the L4 dorsal root. 3. Although no particular abnormalities were found in other types of cutaneous mechanoreceptors, an unusual type of mechanoreceptor was found to be innervating the neuropathic foot. The response characteristics of this type of receptor resemble those of rapidly adapting mechanoreceptors (RAs), but with low and irregular static discharges during a maintained mechanical stimulus. We termed this unusual type as a "modified rapidly adapting" mechanoreceptor (MRA). 4. The response characteristics of MRAs change to those of typical RAs after a systemic injection of phentolamine, an alpha-adrenergic receptor blocker. 5. We conclude that many RAs become abnormal under the influence of sympathetic efferents in neuropathic pain, so that their response patterns change to those of MRAs. We propose that this abnormality is responsible for signaling the mechanical allodynia that can be seen in neuropathic pain states such as causalgia. PMID- 8410154 TI - Multifunctional properties of ambiguous neurons identified electrophysiologically during vocalization in the awake monkey. AB - 1. The nucleus ambiguus (NA) of the medulla contains motoneurons (MNs) for muscles of the larynx, palate, pharynx, and esophagus. Previous studies in anesthetized animals have demonstrated neural discharge correlated closely with respiration, swallowing, and electrical stimulation-elicited vocalization. A preliminary study confirming the above findings was done by recording NA motoneuron extracellular potentials from awake vocalizing monkeys. The present study was undertaken to quantitatively describe discharge properties of a large number of NA neurons recorded from awake, vocalizing monkeys (Macaca nemestrina). 2. In monkeys trained to vocalize for a food reward, extracellular recordings of neurons in and around the nucleus ambiguus were correlated with laryngeal electromyographic activity during vocalization. A nerve cuff electrode on the ipsilateral recurrent laryngeal nerve allowed identification of laryngeal MNs by antidromic activation of laryngeal MNs and the collision test. 3. Most laryngeal MNs became active 100-200 ms before vocalization. They ceased discharging during or immediately after vocalization. Some MNs discharged in close synchrony with bursts of EMG associated with variations in the vocalization. The mean discharge rate of MNs during vocalization was 18 Hz, and the maximum rate in many cells was over 100 Hz. MNs were also active during swallowing. One MN was related only to respiration and one exclusively to swallowing. 4. Some non-motoneurons (Non-MNs) and cells that may possibly be MNs (PossMNs), recorded in and near the NA, showed properties similar to MNs. Many (147) were active only with vocalization, whereas others were active with swallowing (23); respiration (9); vocalization and swallowing (32); vocalization and respiration (40); or vocalization, swallowing, and respiration (17). 5. The present study demonstrates the importance of studying laryngeal MNs in the chronic preparation. Namely, it is shown that both MNs and Non-MNs of the NA are active with more than one activity. Moreover, some Non-MNs are active for only one activity, e.g., vocalization or swallowing. These findings imply the existence of subsets of medullary neurons involved in multiple behaviors for control of generalized laryngeal functions and other subsets related to specific behaviors. PMID- 8410155 TI - Differential modulation of voltage-dependent currents in Hermissenda type B photoreceptors by serotonin. AB - 1. In identified photoreceptors in the eyes of Hermissenda, serotonin (5-HT) enhances the peak and plateau phases of light-evoked generator potentials and modulates light-dependent and voltage-dependent currents. In addition, electrophysiological and morphological studies indicate that 5-HT may contribute to cellular plasticity detected in the visual system of Hermissenda produced by classical conditioning procedures. With the use of conventional two-electrode voltage-clamp techniques, we examined the effects of 5-HT on three distinct currents recorded across the soma membranes of identified lateral and medial type B photoreceptors in the isolated circumesophageal nervous system. 2. The inward rectifier (Iir), a putative K(+)-dependent conductance, activates in 0-Na artificial seawater at membrane potentials greater than -60 mV, is voltage dependent, noninactivating, and reaches steady-state within 800 ms to 3 s at -100 mV. Bath application of 10(-4) M 5-HT consistently enhanced the magnitude of Iir at all potentials tested (-60 to -100 mV) and, in some cases, allowed expression of Iir, which was not initially detectable before the application of 5-HT. 3. The major component of outward K+ current in type B photoreceptors with IA and IK(Ca) blocked is the delayed rectifier [IK(v)]. 5-HT (10(-4) M) produced both an increase as well as a decrease in the peak amplitude of IK(v) and consistently slowed its inactivation rate and reduced twin-pulse inactivation. 4. A previously identified outward K+ current in type B photoreceptors has properties that are similar to the A current (IA).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410156 TI - The location and mechanism of electromotility in guinea pig outer hair cells. AB - 1. The microchamber method was used to examine the motile responses of isolated guinea pig outer hair cells to electrical stimulation. In the microchamber method, an isolated cell is drawn partway into a suction pipette and stimulated transcellularly. The relative position of the cell in the microchamber is referred to as the exclusion fraction. 2. The length changes of the included and excluded segments were compared for constant sinusoidal stimulus amplitude as functions of the exclusion fraction. Both included and excluded segments showed maximal responses when the cell was excluded approximately halfway. Both segments showed smaller or absent responses when the cell was almost fully excluded or almost fully included. 3. When the cell was near to, but not at, the maximum exclusion, the included segment response amplitude was zero, whereas the excluded segment response amplitude was nonzero. In contrast, when the cell was nearly fully included, the excluded segment response amplitude was zero, but the included segment response amplitude was still detectable. A simple model of outer hair cell motility based on these results suggests that the cell has finite resistance terminations and that the motors are restricted to a region above the nucleus and below its ciliated apex (cuticular plate). 4. The function describing length change as a function of command voltage was measured for each segment as the exclusion fraction was varied. The functions were similar at midrange exclusions (i.e., when the segments were about equal length), showing nonlinearity and saturability. The functions were strikingly different when the segment lengths were different. The effects of exclusion on the voltage to length change functions suggested that the nonlinearity and saturability are local properties of the motility mechanism. 5. The diameter changes of both segments were examined. The segment diameter changes were always antiphasic to the length changes. This finding implies that the motility mechanism has an active antiphasic diameter component. The diameter change amplitude was a monotonically increasing function of exclusion for the included segment, and a decreasing function for the excluded segment. 6. The voltage to length-change and voltage to diameter-change functions were measured for the same cell and exclusion fraction. The voltage to diameter-change function was smaller in amplitude than the voltage to length-change function. The functions were of opposite polarity to each other, but were otherwise similar in character. Thus it is likely that the same motor mechanism is responsible for both axial and diameter deformations. PMID- 8410157 TI - Fixation cells in monkey superior colliculus. I. Characteristics of cell discharge. AB - 1. We studied the role of the superior colliculus (SC) in the control of visual fixation by recording from cells in the rostral pole of the SC in awake monkeys that were trained to perform fixation and saccade tasks. 2. We identified a subset of neurons in three monkeys that we refer to as fixation cells. These cells increased their tonic discharge rate when the monkey actively fixated a visible target spot to obtain a reward. This sustained activity persisted when the visual stimulation of the target spot was momentarily removed but the monkey was required to continue fixation. 3. The fixation cells were in the rostral pole of the SC. As the electrode descended through the SC, we encountered visual cells with foveal and parafoveal receptive fields most superficially, saccade-related burst cells with parafoveal movement fields below these visual cells, and fixation cells below the burst cells. From this sequence in depth, the fixation cells appeared to be centered in the deeper reaches of the intermediate layers, and this was confirmed by small marking lesions identified histologically. 4. During saccades, the tonically active fixation cells showed a pause in their rate of discharge. The duration of this pause was correlated to the duration of the saccade. Many cells did not decrease their discharge rate for small-amplitude contraversive saccades. 5. The saccade-related pause in fixation cell discharge always began before the onset of the saccade. The mean time from pause onset to saccade onset for contraversive saccades and ipsiversive saccades was 36.2 and 33.0 ms, respectively. Most fixation cells were reactivated before the end of contraversive saccades. The mean time from saccade terminatioN to pause end was 2.6 ms for contraversive saccades and 9.9 ms for ipsiversive saccades. The end of the saccade-related pause in fixation cell discharge was more tightly correlated to saccade termination, than pause onset was to saccade onset. 6. After the saccade-related pause in discharge, many fixation cells showed an increased discharge rate exceeding that before the pause. This increased postsaccadic discharge rate persisted for several hundred milliseconds. 7. The discharge rate of fixation cells was not consistently altered when the monkey actively fixated targets requiring different orbital positions. 8. Fixation cells discharged during smooth pursuit eye movements as they did during fixation. They maintained a steady tonic discharge during pursuit at different speeds and in different directions, provided the monkey looked at the moving target.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410158 TI - Fixation cells in monkey superior colliculus. II. Reversible activation and deactivation. AB - 1. We tested the hypothesis that a subset of neurons, which we have referred to as fixation cells, located within the rostral pole of the monkey superior colliculus (SC) controls the generation of saccadic eye movements. We altered the activity of these neurons with either electrical stimulation or GABAergic drugs. 2. An increase in the activity of fixation cells in the rostral SC, induced by a train of low-frequency electrical stimulation, delayed the initiation of saccades. With bilateral stimulation the monkey was able to make saccades only after stimulation ceased. 3. Pulses of stimulation delivered during the saccade produced an interruption of the saccade in midflight. The latency to the onset of this perturbation was as short as 12 ms. 4. Injection of the gamma-aminobutyric acid (GABA) antagonist bicuculline into the rostral pole of the SC, which decreases normal GABA inhibition and increases cell activity, increased the latency of saccades to both visual and remembered targets. 5. Injection of the GABA agonist muscimol into the rostral SC, which increases normal GABA inhibition and decreases activity, reduced the latency for saccades to visual targets. The monkey also had difficulty maintaining visual fixation and suppressing unwanted saccades. 6. After muscimol injections, monkeys frequently made very short latency saccades forming a peak in the saccade latency histogram at < 100 ms. These saccades are similar to express saccades made by normal monkeys. This finding suggests that the fixation cells in the rostral SC are critical for controlling the frequency of express saccades. 7. These results support the hypothesis that fixation cells in the rostral SC inhibit the generation of saccadic eye movements and that they form part of a system of oculomotor control, that of visual fixation. PMID- 8410159 TI - Inward rectification and its effects on the repetitive firing properties of bulbospinal neurons located in the ventral part of the nucleus tractus solitarius. AB - 1. An in vitro brain stem slice from adult guinea pigs was used to study the effects of membrane hyperpolarization in two classes of bulbospinal neurons, called types I and II, from the ventral parts of the nucleus tractus solitarius (vNTS). These bulbospinal neurons project to the phrenic motor nucleus and make up the dorsal respiratory group, a sensorimotor integrating area for rhythmic breathing movements. 2. Negative current injections (1 s long) were used in the discontinuous current-clamp mode to study the input resistance (Rin) in both classes of bulbospinal vNTS neurons. The mean Rin for type I neurons was 88.7 +/- 13.8 (SD) M omega (n = 19) and for type II neurons was 92.6 +/- 14.0 M omega (n = 16). Both classes of neurons displayed a depolarizing sag and inward rectification during negative current injections to membrane-potential levels less than or equal to -70 mV. The magnitude of the depolarizing sag became larger as the size of the negative current step was increased. On release from hyperpolarization, both cell types also exhibited a large anode break hyperpolarization (ABH). 3. The ABH was abolished in the presence of 5 mM 4-amino pyridine (4-AP), whereas the depolarizing sag and inward rectification were not affected. In the place of the ABH, a small postinhibitory rebound (PIR) depolarization was observed on release from hyperpolarization. The magnitude of PIR was dependent on the size of the depolarizing sag. In the presence of both 5 mM 4-AP and 5 mM Cs+, the depolarizing sag and PIR were completely blocked, whereas Rin was increased. 4. The ionic currents underlying the ABH and depolarizing sag were directly observed by the use of the discontinuous single electrode voltage-clamp technique. The ABH was caused by activation of an A current (IKA). The depolarizing sag was associated with a hyperpolarization activated inward current (IH), which was activated at membrane-potential levels less than or equal to -70 mV. The peak amplitude of IH in type I neurons was -335 +/- 16 pA (n = 13) and in type II cells was -327 +/- 14 pA (n = 11). 5. IH currents did not display inactivation during the hyperpolarizing voltage step. The IH current became larger when [K+]o was increased from 4 mM (control) to 12 mM and was blocked in the presence of 5 mM Cs+. The estimated reversal potential for the IH current was -41.5 +/- 4.8 mV (n = 8).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410160 TI - Enhancement of synaptic facilitation during the progression of kindling epilepsy by amygdala stimulations. AB - 1. A quantitative analysis of facilitation during the kindling stimulation to the amygdala was conducted by measuring the area between the excitatory potential and the baseline in the averaged tetanic response recorded at the entorhinal cortex. The changes in facilitation were then compared with the development of electrographic afterdischarges (AD) and behavioral seizures in response to successive kindling stimulations. 2. Kindling train pulses (n = 99 or 100; duration: 0.5 ms; frequency: 10 Hz; intensity: AD threshold) were applied to conscious rats until at least one generalized seizure occurred or until 13 stimuli were delivered. 3. Facilitation of the entorhinal responses by kindling stimulation first occurred in the monosynaptic excitatory component and was then followed by a progressive increase in the polysynaptic component that was manifested as the later negative peaks. A clear progressive enhancement was observed in the facilitation by successive kindling stimulations, which also induced prolongation of the AD duration and progression of the seizure stages, indicating that activity-dependent enhancement of facilitation (EF) occurred during the progression of kindling epilepsy. 4. Quantitative analysis revealed that the EF that occurred with the progression of seizure stages was statistically significant (P < 0.001, Friedman test). The AD duration (r = 0.89) and the long-term potentiation (r = 0.85) of the entorhinal responses by single test amygdala stimuli showed a very good linear relation to the EF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410161 TI - Adenosine modulates voltage-gated Ca2+ channels in adult rat sympathetic neurons. AB - 1. Ca(2+)-channel modulation by adenosine was investigated in enzymatically dispersed adult rat superior cervical ganglion (SCG) neurons using the whole-cell variant of the patch-clamp technique. 2. Adenosine produced a concentration dependent decrease in the Ca(2+)-current amplitude with an EC50 of 174 nM and maximum inhibition of 36%. The effects of adenosine on the Ca2+ current were both time and voltage dependent. The inhibition was maximal at +10 mV and decreased at either hyperpolarizing or depolarizing potentials. 3. The inhibitory response desensitized after prolonged (> 1 min) exposure to 10 microM adenosine, whereas multiple brief (< 30 s) applications slightly decreased the subsequent response. 4. Adenosine-induced Ca2+ current inhibition was mediated by an A1-type adenosine receptor, because the half-maximal inhibition value for an A1 receptor selective agonist, chloro-N-cyclopentyladenosine, was 1,000-fold lower than that for an A2 receptor selective agonist, 2-p-(2-carboxyethyl)phenethylamino-5'-N ethylcarbozamido adenosine hydrochloride (33 nM vs. 40 microM, respectively). 5. A guanine nucleotide binding protein (G protein) appeared to be involved in the action of adenosine, because: 1) the adenosine-induced current inhibition could be largely relieved by depolarizing voltage prepulses; 2) tail current analysis revealed that adenosine shifted Ca(2+)-channel activation to more depolarized potentials; and 3) adenosine inhibition was abolished by 2 mM intracellular guanosine 5'-O-(2-thiodiphosphate) or 500 ng/ml pertussis toxin pretreatment. 6. Adenosine did not appear to inhibit L-type Ca2+ channels, because the prolonged tail current component induced by the dihydropyridine "agonist" 2,6-dimethy-3 carbomethoxy-5-nitro-4-(2-trifluoromethyl-phenyl)- 1,4-dihydropyridine (2 microM) was not affected by adenosine. 7. Adenosine-induced inhibition was reduced to approximately 15% after application of 10 microM omega-conotoxin GVIA, suggesting that adenosine primarily inhibits N-type Ca2+ channels. The Ca(2+)-current component resistant to omega-conotoxin GVIA was also resistant to omega-agatoxin IVA (200 nM), suggesting a lack of P-type of Ca2+ channels in SCG neurons. 8. In conclusion, adenosine produces a dose-, time-, and voltage-dependent inhibition of Ca2+ currents in SCG neurons. Adenosine acts on an A1 adenosine receptor subtype in SCG neurons via a pertussis toxin-sensitive G protein to inhibit N type Ca2+ channels and an unidentified Ca(2+)-current component. Modulation of Ca2+ currents by adenosine may be an important mechanism for its inhibitory effect on neurotransmitter release in sympathetic neurons. PMID- 8410162 TI - Hindlimb flexion withdrawal evoked by noxious heat in conscious rats: magnitude measurement of stimulus-response function, suppression by morphine and habituation. AB - 1. The aim of this study was to develop a quantitative behavioral model of nociception. Electromyographic (EMG) recordings from a hamstring flexor muscle provided a measure of the magnitude of hindlimb withdrawals elicited by brief, graded noxious contact thermal stimuli applied to the hindpaw in conscious rats. 2. The magnitude of limb withdrawals showed a significant, usually linear, increase with stimulus temperature from a threshold of approximately 40 up to 52 degrees C. Stimulus-response functions of withdrawal magnitude versus temperature were reproducible within and across rats. Withdrawal magnitude was much more tightly correlated with stimulus temperature (r2 = 0.76, 0.73) than was withdrawal latency (r2 = 0.57, 0.55). 3. Systemic administration of the opiate analgesic morphine (3.5 mg/kg ip) suppressed withdrawals in a naloxone-reversible manner, such that the slope of the stimulus-response function was significantly reduced without an increase in threshold. 4. Successive withdrawals to repeated, identical noxious heat stimuli decreased in a manner consistent with habituation. The response recovered to the prehabituated level after a 15-min rest period, and subsequently decremented even more quickly. The decrement in withdrawal magnitude was greater at lower stimulus intensities and shorter interstimulus intervals, and transferred to a nearby (7.5 mm) but not distant (2.5 cm) site. Evidence for dishabituation was also obtained. 5. The advantages of this method as an animal model of nociception are presented and discussed in terms of the underlying neural circuitry and its modulation. PMID- 8410163 TI - Rat tail flick reflex: magnitude measurement of stimulus-response function, suppression by morphine and habituation. AB - 1. To quantitatively investigate a nocifensive behavioral response, we developed a method to measure the magnitude of the rat's tail flick reflex and its modulation. A radial array of force transducers measured forces of tail flicks (in rostral, horizontal, and vertical planes) elicited by graded noxious radiant thermal stimulation of the conscious rat's tail, from which the overall movement vector was calculated. 2. The rostrally directed component of tail flicks was always larger than dorsal or horizontal components; the latter was usually in a preferred (left or right) direction regardless of which side of the tail was heated. Tail flick force vectors increased from 40 to 46-52 degrees C and then leveled off. Stimulus-response functions were reproducible within and across rats and were fitted by second-order polynomial functions, whose correlation coefficients were similar when the left or right side of the tail was stimulated in separate sessions (r2 = 0.408 and 0.451, respectively). The inverse latency of tail flicks also increased with temperature in a manner fitted by a second-order polynomial (r2 = 0.707, 0.553 for left and right side, respectively). 3. Systemic administration of morphine (1 or 2 mg/kg ip) usually suppressed tail flicks in an all-or-none manner; i.e., flicks at all stimulus temperatures were either totally abolished (n = 7) or unaffected (n = 5) after morphine. In three rats, 1 mg/kg morphine suppressed tail flick magnitude subtotally, reducing the slope of the linear portion of the stimulus-response function. Morphine effects were reversed by the opiate antagonist naloxone. 4. Tail flick magnitude decreased over repeated trials of 44 degrees C heat stimuli delivered to one tail site, recovered after a 15-min rest period, and decremented more quickly with subsequent stimulus repetition. The decrement was less at long (2 or 4 min) than at short (1 min) interstimulus intervals, and high (50 degrees C) than at low (44 degrees C) stimulus intensities. The reflex decrement transferred to a nearby stimulus site in some rats, and was "dishabituated" after a noxious tail pinch. These observations are consistent with habituation of the tail flick reflex. 5. This method, therefore, provides a quantitative and reproducible measure of tail flick reflex magnitude that is sensitive to morphine. The underlying neural circuitry of the tail flick reflex is discussed in relation to limb withdrawal reflexes. PMID- 8410164 TI - Information encoding and the responses of single neurons in the primate temporal visual cortex. AB - 1. The possibility of temporal encoding in the spike trains of single neurons recorded in the temporal lobe visual cortical areas of rhesus macaques was analyzed with the use of principal component and information theory analyses of smoothed spike trains. The neurons analyzed had responses selective for faces. 2. Provided that a correction was applied to earlier methods of principal component analysis used for neuronal spike trains, it was shown that the first principal component provides by a great extent the most information, with the second and third adding only small proportions (on average 18.8 and 8.4%, respectively). 3. It was shown that the magnitude of the second and higher principal components is even smaller if the spike train analysis is started after the onset of the neuronal response, instead of before the neuronal response has started. This suggests that variations in response latency are at least a part of what is reflected by the second and higher principal components. 4. The first principal component was correlated with the mean firing rate of the neurons. The second and higher principal components reflected at least partly the onset properties of the neuronal responses, such as response latency differences between the stimuli. 5. A considerable proportion of the information available from principal components 1-3 is available in the firing rate of the neuron. 6. Periods of the firing rate of as little as 50 or even 20 ms are sufficient to give a reasonable estimate of the firing rate of the neuron. 7. Information theory analysis showed that in short epochs (e.g., 50 ms) the information available from the firing rate can be as high, on average, as 84.4% of that available from the firing rate calculated over 400 ms, and 52.0% of that available from principal components 1-3 in the 400 ms period. It was also found that 44.0% of the information calculated from the first three principal components is available in the firing rates calculated over epochs as short as 20 ms. 8. More information was available near the start of the neuronal response, and the information available from short epochs became less later in the neuronal response. 9. Taken together, these analyses provide evidence that a short period of firing taken close to the start of the neuronal response provides a reasonable proportion of the total information that would be available if a long period of neuronal firing (e.g., 400 ms) were utilized to extract it, even if temporal encoding were used.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410165 TI - Sensitivity of auditory nerve fibers to spectral notches. AB - 1. Listeners use direction-dependent spectral cues introduced by the torso, head, and pinnae to localize the source of a sound in space. Among the prominent direction-dependent spectral features in the free field-to-eardrum transfer function are narrow regions of low acoustic energy referred to as spectral notches. In this paper, we studied the sensitivity of single auditory nerve fibers in the barbiturate-anesthetized cat to broadband noise that had been filtered by a function whose shape approximated natural notches in the free field to-eardrum transfer function. 2. Two experimental paradigms were employed. The first was the repeated presentation of a burst of broadband noise filtered by the simulated-notch function. Center frequency of the notch was held constant at or around the fiber characteristic frequency (CF). We refer to this as a "stationary" notch stimulus. The second paradigm was the repeated presentation of a broadband noise that was constructed from noise segments, each filtered by the simulated notch, whose CF was incremented and then decremented in a systematic way. We refer to this as a "moving" notch stimulus. Results from these two paradigms were compared with respect to notch detection. 3. Data were obtained from 161 single auditory nerve fibers having CFs ranging from 0.4 to 40 kHz. Most fibers studied had CFs > 5 kHz, and they detected the presence of the spectral notch in an intensity- and frequency-dependent manner. Each fiber responded vigorously to the presence of broadband noise. When the CF of the notch encroached on the response area of the fiber, there was a demonstrable reduction in discharge rate. The greatest reduction in discharge rate occurred when the notch was centered at the fiber's CF and when the level of the notch signal was some 15-55 dB above the fiber's noise threshold. There was close association in the frequency-intensity plane between the position of the most effective notch and the fiber's threshold tuning curve. 4. For high-spontaneous rate fibers, a moving-notch stimulus, but not a stationary one, reduced the discharge below the spontaneous rate at and in the immediate vicinity of the most effective notch frequency. This increases sensitivity to a spectral notch and suggests a mechanism by which localization ability is enhanced when there is relative motion between a sound source and the head.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410166 TI - Virtual-space receptive fields of single auditory nerve fibers. AB - 1. Sounds reaching the tympanic membranes are first modified by the acoustic properties of the torso, head, and external ear. For certain frequencies in the incident sound there results a complex, direction-dependent spatial distribution of sound pressure at the eardrum such that, within a sound field, localized areas of pressure maxima are flanked by areas of pressure minima. Listeners may use these spatial maxima and minima in localizing the source of a sound in space. The results presented describe how information about this spatial pressure pattern is transmitted from the cochlea to the central auditory system via single fibers of the auditory nerve. 2. Discharges of single fibers of the auditory nerve were studied in Nembutal-anesthetized cats [characteristic frequencies (CFs) ranged from 0.4 to 40 kHz]. Click stimuli were derived from sound-pressure waveforms that were generated by a loudspeaker placed at 1,800 locations around the cat's head and recorded at the tympanic membrane with miniature microphones. Recorded signals were converted to acoustic stimuli and delivered to the ear via a calibrated and sealed earphone. The full complement of signals is referred to as "virtual acoustic space," and the spatial distribution of discharges to this array of signals is referred to as a "virtual-space receptive field" (VSRF). 3. Fibers detect both pressure maxima and pressure minima in virtual acoustic space. Thus VSRFs take on complex shapes. 4. VSRFs of fibers of the same or similar CF having low spontaneous rates had the same overall pattern as those from high spontaneous rate (HSR) fibers. For HSR fibers, the VSRF is obscured by the high background spike activity. 5. Comparison of the VSRF and isolevel contour maps of the stimulus derived at various frequencies revealed that auditory nerve fibers most accurately extract spectral information contained in the stimulus at a frequency close to or slightly higher than CF. PMID- 8410167 TI - Relationships between color, shape, and pattern selectivities of neurons in the inferior temporal cortex of the monkey. AB - 1. To examine the way in which information from different visual submodalities is integrated in the inferior temporal (IT) cortex of the monkey, we studied the relationships between the color, shape, and pattern selectivities of individual neurons in IT cortex of two awake macque monkeys. Neurons were recorded while each animal performed a visual fixation task. For each neuron, we analyzed selectivity for the visual submodalities of color, contour shape, and textural pattern using preselected standard sets of visual stimuli, namely colored geometrical shapes with certain patterns. 2. About two thirds (62%) of single neurons whose activities were recorded from the anterior part of IT cortex, which included the ventral bank of the superior temporal sulcus, responded to one or more of the stimuli in the standard sets of stimuli used. An index (stimulus selectivity index) was calculated for each neuron to quantify how well a cell discriminated the preferred stimulus from the least preferred stimulus in each set of stimuli. The stimulus selectivity index, as well as the statistical significance of the variation in the responses to the stimuli within a given set, was used to classify a cell as selective or not selective in a given submodality. Of the neurons whose responses were analyzed quantitatively, 69% were selective for color, 68% were selective for shape, and 82% were selective for pattern. 3. Of the neurons that were tested with respect to the selectivity for color and for shape, 45% were selective both for color and shape, 50% were selective for either color or shape, and only 5% were not selective for color or shape. These frequencies were not significantly different from those predicted from the occurrence of the selective and nonselective neurons in each submodality if the independence of the selectivities for color and shape is assumed. We also found that the color preference of individual neurons does not depend on the shape of the stimulus. These results indicate that there was no overt interaction between the selectivities for color and shape in these IT neurons. 4. Of the neurons that were compared with respect to the selectivity for color and for pattern, 58% were selective both for color and pattern, 38% were selective for either color or pattern, and only 4% were not selective for color or pattern. There was no correlation between the degree of color selectivity and the degree of pattern selectivity of individual neurons.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410168 TI - A caffeine- and ryanodine-sensitive Ca2+ store in avian sensory neurons. AB - 1. We identified and studied the function of ryanodine receptors in neurons isolated from dorsal root ganglia (DRG) of 10-day-old chick embryos. 2. A monoclonal antibody (mAb 34C) that recognizes all known ryanodine receptor isoforms in skeletal and cardiac muscle and CNS identified ryanodine receptor like immunoreactivity in cultured DRG neurons. 3. Using the permeabilized patch technique to record membrane currents, we found that calcium currents were followed by a current with characteristics of a Ca(2+)-activated Cl- current (ICl(Ca)) in approximately two-thirds of the neurons. In these cells, acute application of 10 mM caffeine activated a similar ICl(Ca) and this effect was inhibited by 10 microM ryanodine. The activation of ICl(Ca) by caffeine was not dependent on extracellular Ca2+. These data suggest that caffeine raises intracellular free Ca2+ (Cai2+) by activating the release of Ca2+ from an intracellular store and that this Ca2+ activates the membrane conductance responsible for ICl(Ca). 4. The magnitude of ICl(Ca) activated by depolarization was not affected by ryanodine, implying that the Ca2+ that activates ICl(Ca) in this protocol is supplied by the Ca2+ current without amplification by a ryanodine-sensitive mechanism such as Ca(2+)-induced Ca2+ release. 5. We also used indo-1 to measure Cai2+ in DRG neurons. Ten millimolar caffeine caused a transient increase in Cai2+ that was inhibited by 10 microM ryanodine. 6. The ability of caffeine to elevate Cai2+ and activate ICl(Ca) was reduced at higher temperatures, suggesting increased Ca2+ sequestration. 7. These data demonstrate the existence of an intracellular store of Ca2+ that can be mobilized by a caffeine- and ryanodine-sensitive mechanism. The release of Ca2+ from this store can elevate Cai2+ and modulate membrane conductances. PMID- 8410169 TI - Projection from the sensory to the motor cortex is important in learning motor skills in the monkey. AB - 1. The projection from the somatosensory cortex to the primary motor cortex has been proposed to play an important role in learning novel motor skills. This hypothesis was examined by studying the effects of lesions to the sensory cortex on learning of new motor skills. 2. We used two experimental paradigms to reveal the effects of lesions on learning of new motor skills. One task was to catch a food pellet falling at various velocities. The other task was to catch a food pellet from a rotating level. Both tasks required acquisition of novel motor skills. 3. The training was started after a lesion of the hand area in the somatosensory cortex of one hemisphere. In both tasks, monkeys had severe difficulty in learning the new skills with the hand contralateral to the ablated somatosensory cortex, compared with the hand contralateral to the intact hemisphere. 4. After acquisition of the motor skill in the hand contralateral to intact hemisphere, lesion of the somatosensory cortex hand area did not abolish the learned motor skill. 5. In control experiments, monkeys were trained to pick up a food pellet from a rotating board. This task did not necessitate acquisition of new motor skills, but could be performed by utilizing existing motor skills. Lesion in the somatosensory cortex before or after the training did not affect the execution of this task by either hand. 6. It is concluded that the corticocortical projection from the somatosensory to the motor cortex plays an important role in learning new motor skills, but not in the execution of existing motor skills. PMID- 8410170 TI - Responsiveness to D-glucose in neurosecretory cells of crustaceans. AB - 1. An electrophysiological study of the D-glucose sensitivity of X-organ (XO) neurosecretory cell bodies in crayfish was carried out with the use of microelectrodes, perforated, and cell-attached patch-clamp techniques. 2. Glucose depolarizes the membrane potential of XO cells in a concentration-dependent manner. 3. Depolarization produced by glucose initiates a change in the pattern of electrical activity. Silent cells began to discharge action potentials. When bursting cells are depolarized by glucose, their action potentials are no longer grouped in bursts or disappear entirely. 4. Although the membrane potential returns to its initial value after removing glucose from the bath, discharge patterns of the cells may remain different. This suggests that besides the depolarizing effect, once the cells have been exposed to glucose, the sugar switches on a process that is maintained for a long time. 5. Glucose produced a reduction of membrane steady-state conductance, and a shift of reversal potential of membrane currents to a more positive value. 6. Depolarization induced by D glucose appears to be related with a closure of potassium channels. 7. Glucose effect was thought to be generated by a product of metabolism that would act as intracellular mediator. PMID- 8410171 TI - Fictive swimming elicited by electrical stimulation of the midbrain in goldfish. AB - 1. We developed a fictive swimming preparation of goldfish that will allow us to study the cellular basis of interactions between swimming and escape networks in fish. 2. Stimulation of the midbrain in decerebrate goldfish produced rhythmic alternating movements of the body and tail similar to swimming movements. The amplitude and frequency of the movements were dependent on stimulus strength. Larger current strengths or higher frequencies of stimulation produced larger amplitude and/or higher-frequency movements. Tail-beat frequency increased roughly linearly with current strength over a large range, with plateaus in frequency sometimes evident at the lowest and highest stimulus strengths. 3. Electromyographic (EMG) recordings from axial muscles on opposite sides at the same rostrocaudal position showed that stimulation of the midbrain led to alternating EMG bursts, with bursts first on one side, then the other. These bursts occurred at a frequency equal to the tail-beat frequency and well below the frequency of brain stimulation. EMG bursts recorded from rostral segments preceded those recorded from caudal segments on the same side of the body. The interval between individual spikes within EMG bursts sometimes corresponded to the interval between brain stimuli. Thus, whereas the frequency of tail beats and EMG bursts was always much slower than the frequency of brain stimulation, there was evidence of individual brain stimuli in the pattern of spikes within bursts. 4. After paralyzing fish that produced rhythmic movement on midbrain stimulation, we monitored the motor output during stimulation of the midbrain by using extracellular recordings from spinal motor nerves. We characterized the motor pattern in detail to determine whether it showed the features present in the motor output of swimming fish. The fictive preparations showed all of the major features of the swimming motor pattern recorded in EMGs from freely swimming fish. 5. The motor nerves, like the EMGs produced by stimulating midbrain, showed rhythmic bursting at a much lower frequency than the brain stimulus. Bursts on opposite sides of the body alternated. The frequency of bursting ranged from 1.5 to 13.6 Hz and was dependent on stimulus strength, with higher strengths producing faster bursting. Activity in rostral segments preceded activity in caudal ones on the same side of the body. Some spikes within bursts of activity occurred at the same frequency as the brain stimulus, but individual brain stimuli were not as evident as those seen in some of the EMGs. 6. The duration of bursts of activity in a nerve was positively and linearly correlated with the time between successive bursts (cycle time).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410172 TI - Voltage- and space-clamp errors associated with the measurement of electrotonically remote synaptic events. AB - 1. The voltage- and space-clamp errors associated with the use of a somatic electrode to measure current from dendritic synapses are evaluated using both equivalent-cylinder and morphologically realistic models of neuronal dendritic trees. 2. As a first step toward understanding the properties of synaptic current distortion under voltage-clamp conditions, the attenuation of step and sinusoidal voltage changes are evaluated in equivalent cylinder models. Demonstration of the frequency-dependent attenuation of voltage in the cable is then used as a framework for understanding the distortion of synaptic currents generated at sites remote from the somatic recording electrode and measured in the voltage clamp recording configuration. 3. Increases in specific membrane resistivity (Rm) are shown to reduce steady-state voltage attenuation, while producing only minimal reduction in attenuation of transient voltage changes. Experimental manipulations that increase Rm therefore improve the accuracy of estimates of reversal potential for electrotonically remote synapses, but do not significantly reduce the attenuation of peak current. In addition, increases in Rm have the effect of slowing the kinetics of poorly clamped synaptic currents. 4. The effects of the magnitude of the synaptic conductance and its kinetics on the measured synaptic currents are also examined and discussed. The error in estimating parameters from measured synaptic currents is greatest for synapses with fast kinetics and large conductances. 5. A morphologically realistic model of a CA3 pyramidal neuron is used to demonstrate the generality of the conclusions derived from equivalent cylinder models. The realistic model is also used to fit synaptic currents generated by stimulation of mossy fiber (MF) and commissural/associational (C/A) inputs to CA3 neurons and to estimate the amount of distortion of these measured currents. 6. Anatomic data from the CA3 pyramidal neuron model are used to construct a simplified two-cylinder CA3 model. This model is used to estimate the electrotonic distances of MF synapses (which are located proximal to the soma) and perforant path (PP) synapses (which are located at the distal ends of the apical dendrites) and the distortion of synaptic current parameters measured for these synapses. 7. Results from the equivalent cylinder models, the morphological CA3 model, and the simplified CA3 model all indicate that the amount of distortion of synaptic currents increases steeply as a function of distance from the soma. MF synapses close to the soma are likely to be subject only to small space-clamp errors, whereas MF synapses farther from the soma are likely to be substantially attenuated.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410173 TI - Oscillatory properties of the central pattern generator for locomotion in neonatal rats. AB - 1. The oscillatory properties of the lumbar spinal networks that generate locomotor activity in mammals were studied. These experiments were performed on an in vitro isolated spinal cord preparation from newborn rats. Adding to the saline serotonin (5-HT) and N-methyl-D-aspartate (NMDA) excitatory amino acid receptor agonist (N-methyl-D,L-aspartate, NMA) induced rhythmic locomotor-like activity that was recorded in the ventral roots. 2. The period of the rhythmic locomotor-like activity could be set by combining the neurotransmitters at various concentrations. The combined transmitters also acted on the instantaneous variations of the period value. The stability of the period from one cycle to another increased when 5-HT was mixed with NMA compared with the activity induced by bath application of the compound alone. 3. K(+)-induced depolarizations were used to set the neuron membrane potential. These systematic changes in the K+ concentration resulted in a change in the period value. 4. Stimulation of the dorsal roots reset the ongoing rhythm. Phase-response curves were drawn up that showed that the spinal networks exhibit a differential sensitivity to the same stimulus depending on the phase at which the stimulus is applied. The free running activity could be entrained by the cyclic volleys of sensory discharges. Entrainment occurred, on average, between -28 and 36% of the rest period. Because the lumbar segments were completely isolated by means of a sucrose blockade of all descending neuronal activity, the entrainment of the locomotor activity must take place directly at the lumbar level. 5. In conclusion, our data show that, at birth, the rhythmic activity produced by the spinal networks is based on a neuronal architecture that displays oscillatory properties. At this early age, the peripheral sensory afferents have already established connections with the central pattern generator. These sensory inputs are able to control the cycling activity of the spinal networks step by step. PMID- 8410174 TI - Visual receptive field properties in kitten pretectal nucleus of the optic tract and dorsal terminal nucleus of the accessory optic tract. AB - 1. Neurons in the pretectal nucleus of the optic tract (NOT) and dorsal terminal nucleus of the accessory optic tract (DTN) were recorded in anesthetized and paralyzed kittens on postnatal days 18 to 48 (P18-P48) as well as in adult cats. 2. Spontaneous as well as stimulus driven discharge rates of NOT-DTN neurons in the youngest kittens (P18-P23) are significantly lower than in older kittens (P27 P33) or adult cats. 3. Visual latencies of NOT-DTN neurons in P18-P23 kittens are significantly longer than in P27-P33 kittens. They further decrease as the animals reach adulthood. 4. Already in the youngest animals recorded in this experimental series (P18) NOT-DTN neurons were selective for ipsiversive horizontal stimulus movement. When expressed as the difference between response strength during stimulation in the preferred and the nonpreferred direction, P18 P23 NOT-DTN neurons are less direction selective than NOT-DTN cells in older animals. However, the normalized directional tuning expressed as percent change in discharge rate per degree change in stimulus direction away from the preferred direction (where discharge rate is set 100%) is about equal in all age groups. 5. NOT-DTN neurons in P18-P23 kittens respond to a rather limited range of stimulus speeds with an optimum at approximately 10 degrees/s. In P27-P33 kittens, NOT-DTN neurons increase their responsive range to higher stimulus speeds. As the animals approach adulthood, the range of effective stimulus speeds further broadens to include very low ones. 6. In P18-P23 kittens, the majority of NOT-DTN neurons is exclusively activated by the contralateral eye; only a few neurons receive an additional input from the ipsilateral eye. In P27-P48 kittens, the influence of the ipsilateral eye has significantly increased but with the majority of NOT-DTN cells still being dominated by the contralateral eye. Finally, in adults, a further strengthening of the ipsilateral input leads to a more binocularly balanced input to NOT-DTN cells. 7. Electrical stimulation in areas 17 and 18 did not elicit orthodromic action potentials in NOT-DTN neurons before P27. Thus the cortical input to the NOT-DTN in kittens becomes functional only at 4 wk of age. 8. In conclusion, the significant changes of visual response properties of NOT DTN neurons coincide with the time when the cortical input to the NOT-DTN becomes functional.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410175 TI - Firing behavior of brain stem neurons during voluntary cancellation of the horizontal vestibuloocular reflex. I. Secondary vestibular neurons. AB - 1. The single-unit activity of vestibular neurons was recorded in alert squirrel monkeys. The monkeys had been trained to track a small visual target by generating smooth pursuit eye movements and to cancel their vestibuloocular reflex (VOR) by fixating a head stationary target. The monkeys were seated on a vestibular turntable, and their heads were held in the plane of the horizontal semicircular canals. The responses of 45 type I vestibular neurons whose activity was related to ipsilateral horizontal head movements were recorded. In 19 of 30 cells tested, electrical stimulation (0.1-ms monophasic pulses, < or = 800 microA) of the ipsilateral vestibular nerve evoked a spike at a monosynaptic latency (0.7-1.3 ms). 2. The spiking behavior of each cell was recorded during several behavioral paradigms: 1) spontaneous eye movements, 2) horizontal smooth pursuit of a target that was moved sinusoidally +/- 20 degrees/s at 0.5 Hz, 3) horizontal VOR during 0.5-Hz sinusoidal turntable rotations +/- 40 degrees/s (VORs), and 4) voluntary cancellation of the sinusoidal VOR by fixation of a head stationary target during 0.5-Hz sinusoidal turntable rotation at +/- 40 degrees/s in the light (VORCs). 3. The response of most (34) of the units was recorded during unpredictable 100-ms steps in head acceleration (400 degrees/s2) that were generated while the monkey was fixating a target light. The acceleration steps were generated either when the monkey was stationary (VORt paradigm) or when the turntable was already rotating, and the monkey was canceling its VOR (VORCt paradigm). Smaller eye movements were evoked when the acceleration step was generated during VOR cancellation. 4. Type I vestibular units were grouped into two classes on the basis of the relationship of their firing rate to eye movements. The discharge rate of 20 "pure vestibular" units was not clearly related to eye movements. The remaining 25 units were classified as position vestibular-pause (PVP) neurons. PVP neurons increased their firing rate during contralateral eye movements and during ipsilateral turntable rotations, and paused during saccadic eye movements. 5. Most (17/20) pure vestibular neurons generated the same response to vestibular stimuli when the monkeys canceled their VOR as they did during the VOR in both the sinusoidal and acceleration step paradigms. 6. The head velocity sensitivity of most (19/24) PVP neurons was reduced by 20-60% during VORCs, compared with their response during the VORs. The PVP neurons whose sensitivity of head movements was reduced during VORCs also exhibited a reduced vestibular sensitivity during VORCt.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410176 TI - Firing behavior of brain stem neurons during voluntary cancellation of the horizontal vestibuloocular reflex. II. Eye movement related neurons. AB - 1. The single-unit activity of neurons in the vestibular nucleus, the prepositus nucleus, and the abducens nucleus, whose activity was primarily related to horizontal eye movements, was recorded in alert squirrel monkeys that were trained to track a small visual target by generating smooth pursuit eye movements and to cancel their horizontal vestibuloocular reflex (VOR) by fixating a head stationary target. 2. The spiking behavior of each cell was recorded during 1) spontaneous eye movements, 2) horizontal smooth pursuit of a target that was moved sinusoidally +/- 20 degrees/s at 0.5 Hz, 3) horizontal VOR evoked by 0.5-Hz sinusoidal turntable rotations +/- 40 degrees/s (VORs), and 4) voluntary cancellation of the VOR by fixation of a head-stationary target during 0.5-Hz sinusoidal turntable rotation at +/- 40 degrees/s (VORCs). The responses of most (28/42) of the units were recorded during unpredictable 100-ms steps in head acceleration (400 degrees/s2) that were generated while the monkey was fixating a target light. The acceleration steps were generated either when the monkey was stationary or when the turntable was already rotating (VORt trials), and the monkey was canceling its VOR (VORCt trials). 3. The firing behavior of all 12 of the abducens neurons recorded was closely related to horizontal eye position and eye velocity during all of the behavioral paradigms used, although there was a small but significant increase in the eye position sensitivity of many of these units when the eye was moving (smooth pursuit) versus when the eye was stationary (fixation). 4. Many neurons in the prepositus nucleus and the medial vestibular nucleus (n = 15) were similar to abducens neurons, in that their firing rate was related primarily to horizontal eye position and eye velocity, regardless of the behavioral paradigm used. These cells were, on average, more sensitive to eye position and smooth pursuit eye velocity than were abducens neurons. 5. The firing rate of 15 other neurons in the prepositus and medial vestibular nucleus was related primarily to horizontal smooth pursuit eye movements. The tonic firing rate of all of these smooth pursuit (SP) cells was related to horizontal eye position, and the majority generated bursts of spikes during saccades in all directions but their off direction. Six of the SP neurons fired in phase with ipsilateral eye movements, whereas the remaining nine were sensitive to eye movements in the opposite direction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410177 TI - Increased propensity for nonsynaptic epileptiform activity in immature rat hippocampus and dentate gyrus. AB - 1. Low-[Ca2+] bursting was studied in hippocampal slices from immature and adult rats to test the hypothesis that the increased seizure susceptibility of the immature brain involves nonsynaptic mechanisms. Extracellular recordings were obtained from area CA1 of the hippocampus and from the dentate gyrus in slices from rats 6-9 days old (1 wk), 11-15 days old (2 wk), 19-23 days old (3 wk), and > 60 days old (adult). These slices were exposed to a low-[Ca2+] solution that included the calcium chelator, ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA), and the excitatory amino acid antagonists, 6,7 dinitroquinoxaline-2,3-dione (DNQX) and DL-2-amino-5-phosphonopentanoic acid (AP 5). They were also exposed to a hyposmolar low-[Ca2+] solution (diluted with 20% H2O by volume), which induced or intensified the bursting. The propensity for nonsynaptic bursting and the characteristics of the bursts were compared between age groups. 2. The 1-wk group showed no bursting activity under any treatment condition in either CA1 or the dentate gyrus. Bursting occurred more frequently in the 2- and 3-wk groups than in the adult group in both CA1 and the dentate gyrus. 3. In CA1 the duration of the bursts was longer in the 2- and 3-wk groups as compared with the adult group. The number of population spikes per burst was also higher in slices from immature rats in dilute low-[Ca2+] solution. These findings demonstrate that nonsynaptic bursting in area CA1 is more robust in tissue from immature rats than adults.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410178 TI - Broad directional tuning in spinal projections to the cerebellum. AB - 1. Spinocerebellar neurons that project in the dorsal spinocerebellar tract (DSCT) receive mono- and polysynaptic inputs from specific sensory receptors in the hindlimb, and they project mossy fiber terminals to the cerebellar vermis. We examined the functional organization of these neurons and found that it relates to whole-limb parameters like limb posture and direction of limb movement. 2. We recorded the activity of 444 DSCT units during passive perturbations of the hind foot in anesthetized cats. The movements were either confined a single joint (the ankle; 234 cells) or involved the entire hindlimb (210 cells). The cells exhibited opposite responses for opposite directions of whole-limb movement, but a variety of response patterns for opposite directions of movement at one joint. We interpret the result to imply that the population encodes information about the whole limb rather than single joints. 3. Most of the 78 neurons recorded during passive limb placements (63%) responded to changes in limb length and also changes in limb orientation. In fact, the activity of most of the cells was broadly tuned with respect to the direction of passive limb movements generated by moving the hind foot in the sagittal plane. Changes in unit activity could be described by a cosine tuning function with respect to foot positions (72% of responses) and directions of foot movement (50%). 4. The similarity of this behavior to that of neurons in the motor cortex and cerebellar nuclei recorded during voluntary movements is consistent with a common neural code to represent the sensorimotor parameters of limb movement. PMID- 8410179 TI - Functional grouping of the cortico-pretectal projection. AB - The ascendency of the cerebral cortex in mammals naturally raises questions about the role of the archetypal subcortical centers we share in common with other phyla. Here we report a situation in which an ancient oculomotor control center, the nucleus of the optic tract, is not so much dominated by the cerebral cortex as served by it. We suggest that the organization of cortical output to subcortical centers may be helpful in understanding the function of the cerebral cortex. PMID- 8410180 TI - The direction change concept for reticulospinal control of goldfish escape. AB - This is an analysis of whether biomechanical or kinematic variables are controlled by descending reticulospinal commands to the spinal cord during escape responses (C-starts) in the goldfish. We studied how the animal contracted its trunk musculature to orient an escape trajectory. We used trunk EMG recordings as a measure of the reticulospinal output to the musculature and we simultaneously gathered high-speed cinematic records of the resulting movements. We found that the escape trajectory is controlled by (1) the relative size of the agonist versus the antagonist muscle contractions on two sides of the body and (2) the timing between these contractions. We found no separate signal for forward propulsion (or force) apart from the initial stage 1 bending of the body. Rather, the neural specification of force is embedded in the commands to bend the body. Thus, our findings demonstrate the importance of the angular kinematic components, or direction changes, caused by the descending reticulospinal command. This new direction change concept is important for two reasons. First, it unifies the diversity of C-start movement patterns into a single and rather simple quantitative model. Second, the model is analogous to the systematic EMG and kinematic changes observed by others to underlie single joint movements of limbs in other vertebrates such as primates. As in these cases, the fish capitalizes on the mechanical properties of the muscle by setting the extent and timing of agonist and antagonist contractions. This, plus the fact that sensory feedback is likely to be minimal, may enable the animal to reduce the number of computational steps in its motor commands used to produce the escape response. Because horizontal body movements in fish are a fundamental vertebrate movement pattern produced by a highly conserved brainstem movement system, our findings may have general implications for understanding the neural basis of rapid movements of diverse body parts. PMID- 8410181 TI - Geometry of orientation and ocular dominance columns in monkey striate cortex. AB - In addition to showing that ocular dominance is organized in slabs and that orientation preferences are organized in linear sequences likely to reflect slabs, Hubel and Wiesel (1974a) discussed the intriguing possibility that slabs of orientation might intersect slabs of ocular dominance at some consistent angle. Advances in optical imaging now make it possible to test this possibility directly. When maps of orientation are analyzed quantitatively, they appear to arise from a combination of at least two competing themes: one where orientation preferences change linearly along straight axes, remaining constant along perpendicular axes and forming iso-orientation slabs along the way, and one where orientation preferences change continuously along circular axes, remaining constant along radial axes and forming singularities at the centers of the spaces enclosed. When orientation patterns are compared with ocular dominance patterns from the same cortical regions, quantitative measures reveal (1) that singularities tend to lie at the centers of ocular dominance columns, (2) that linear zones (arising where orientation preferences change along straight axes) tend to lie at the edges of ocular dominance columns, and (3) that the short iso orientation bands within each linear zone tend to intersect the borders of ocular dominance slabs at angles of approximately 90 degrees. PMID- 8410182 TI - The layout of iso-orientation domains in area 18 of cat visual cortex: optical imaging reveals a pinwheel-like organization. AB - In this study we used optical imaging based on activity-dependent intrinsic signals to determine the distribution of cells responding to gratings of various orientations moving in different directions in area 18 of cat visual cortex. To test directional-selective clustering of neurons, we compared cortical activity maps obtained by stimulation with two gratings of identical orientation but moving in opposite directions. We found those maps to be almost identical, suggesting that neurons are not notably clustered into directionality columns. We also compared activity maps obtained with gratings of different orientations. Each of the orientation maps was similar to the 2-deoxyglucose maps previously reported. Having compiled the information obtained from the different orientations into one "orientation preference map," we found, in contrast to earlier reports, that iso-orientation domains are not elongated parallel bands but are small patches organized in "pinwheels" around points that we refer to as "orientation centers." We furthermore show that the only locations at which orientation preference changes rapidly are these orientation centers and not lines or loops. In addition, this report clarifies that our observations on the functional architecture of cat area 18, although at first sight at variance with earlier observations, are actually fully consistent with them. We therefore propose that in cat visual cortex pinwheel-like patterns of orientation preference form an irregular mosaic of modular units with an average density of 1.2 pinwheels per square millimeter. PMID- 8410183 TI - Differential metabolic and electrical activity in the somatic sensory cortex of juvenile and adult rats. AB - We have examined relative levels of metabolic and electrical activity across layer IV in the primary somatic sensory cortex (S1) of the rat in relation to regions of differential postnatal cortical growth. Each of several indices used- mitochondrial enzyme histochemistry, microvessel density, Na+/K+ pump activity, action potential frequency, and deoxyglucose uptake--indicate regional variations of metabolic and electrical activity in this part of the brain in both juvenile (1-week-old) and adult (10-12-week-old) animals. At both ages, areas of the somatic sensory map related to special sensors such as whiskers and digital pads showed evidence of the most intense activity. Thus, mitochondrial enzyme staining, blood vessel density, and Na+/K+ ATPase activity were all greatest in the barrels and barrel-like structures within S1, and least in the adjacent interbarrel cortex and the cortex surrounding S1. Multiunit recordings in and around the posteromedial barrel subfield of anesthetized animals also showed that the average ratio of evoked to spontaneous activity was greater in barrels than in the surrounding, metabolically less active cortex. Furthermore, autoradiograms of labeled deoxyglucose accumulation in awake behaving animals indicated systematic differences in neural activity across S1 barrels and barrel-like structures showed more deoxyglucose accumulation than interbarrel, nonbarrel, or peri-S1 cortex. These regional differences in neural activity correspond to regional differences in neocortical growth (Riddle et al., 1992). The correlation of greater electrical activity, increased metabolism, and enhanced cortical growth during postnatal maturation suggests that neural activity foments the elaboration of circuitry in the developing brain. PMID- 8410184 TI - Growth factors protect PC12 cells against ischemia by a mechanism that is independent of PKA, PKC, and protein synthesis. AB - We have established an in vitro model of ischemia incorporating the combination of anoxia with glucose deprivation, which is toxic to PC12 cells. In this model, nerve growth factor (NGF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF) improve PC12 cell survival. K252a, a specific inhibitor of NGF-induced trk p140 autophosphorylation, did not alter the neuroprotection provided by EGF or bFGF, yet it completely abolished the protection provided by NGF. Activation of protein kinase A (PKA) with dibutyryl-cAMP also protected during ischemia, although it was not additive with the effect provided by growth factors. Furthermore, growth factors protected a PKA-deficient mutant as effectively as the parental cell line; thus, activation of PKA is protective against ischemia but is not necessary for the action of peptide growth factors. Neither the stimulation of protein kinase C (PKC) with acute phorbol ester treatment nor the downregulation of PKC with chronic high-dose phorbol ester treatment resulted in an altered response to growth factors in either the PC12 wild type or PKA-deficient mutant. Thus, protection by peptide growth factors depends on neither PKA nor PKC. Furthermore, downregulation of PKC alone was protective, indicating that PKC may contribute to toxicity. Interestingly, treatment with the kinase inhibitor H-7 was neuroprotective and may have enhanced the neuroprotective effect of NGF. In contrast, staurosporine, a broadly acting kinase inhibitor, inhibited the neuroprotective effect of NGF, but not of EGF or FGF.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410185 TI - Dopamine transporter expression confers cytotoxicity to low doses of the parkinsonism-inducing neurotoxin 1-methyl-4-phenylpyridinium. AB - The uptake of 1-methyl-4-phenylpyridinium (MPP+), the active metabolite of the parkinsonism-inducing neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), was studied in various mammalian cell lines transfected, respectively, with the cloned human and rat dopamine transporters, and compared with rat striatal synaptosome preparations. Only in neuronally derived cell lines such as NG108-15, NS20Y, and SK-N-MC cells did MPP+ have a KM for the cloned transporters comparable to that of dopamine as seen in rat striatal synaptosomes. In non neuronally derived cells such as COS-7, CHO, and Ltk- cells transiently or permanently expressing the transporters, the KM of MPP+ was at least 10-fold higher. The permanent expression of either the cloned human or rat dopamine transporters conferred to SK-N-MC cells susceptibility to the cytotoxic effects of low concentrations of MPP+. The extent of this effect was dependent on the expression level of the dopamine transporters and could be specifically antagonized by the catecholamine uptake inhibitor mazindol. There were no significant differences in the susceptibility to MPP+ of cells expressing similar levels of either the human or rat dopamine transporter. The demonstration for the first time of a quantitative relationship between the cellular expression of the plasma membrane transporter and the extent of the cytotoxic effects of MPP+ suggests that known differences in vulnerability of various brain regions to MPP+ cytotoxicity might be related to their actual content of dopamine uptake sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410186 TI - Axonal guidance defects in a Caenorhabditis elegans mutant reveal cell-extrinsic determinants of neuronal morphology. AB - Mutations in the gene unc-53 of Caenorhabditis elegans result in behavioral and anatomical abnormalities. Immunocytochemistry and electron microscopy revealed neuroanatomical defects in all main longitudinal nervous tracts. Whole tracts were found to be misguided in specific ways suggesting that unc-53 affects pioneering axons. The four lateral microtubule cells (LMs), which are probably pioneering neurons, were examined in greatest detail. In the mutants, the processes of the LMs leave their normal position on the body wall and terminate prematurely. Examination of five unc-53 alleles for penetrance and expressivity of these defects revealed a spatial restriction in the requirement for unc-53. The morphology and positioning of the branch of the posterior lateral microtubule cells (PLMs) were also examined. In wild-type animals, the PLM branches lack the ultrastructural specializations of the main process, which include large microtubules, apposition to the cuticle, and a polarized extracellular matrix (the mantle). Two differences were noted in unc-53 mutants. First, a majority of PLMs branch at random and display an abnormally enlarged branching point and branch cross section. The unusual branch morphologies correlate with branch position, rather than PLM length. Second, the ectopic branches display the specific ultrastructural features characteristic of the main process. Furthermore, after entering the ventral nerve cord, the abnormal branches constantly change position relative to the other processes and the hypodermis, retaining their specialized microtubules throughout, but displaying a mantle only when in direct contact with hypodermis. Taken together, these observations suggest that the differentiated features of the PLMs, including process length, branch position, intracellular branch morphology, and surrounding extracellular matrix, are locally specified by cell-extrinsic cues, some of which require unc 53. PMID- 8410187 TI - Signal transduction events mediated by the BDNF receptor gp 145trkB in primary hippocampal pyramidal cell culture. AB - The trkB gene encodes a tyrosine kinase receptor, gp145trkB, for brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4). To understand the role of gp145trkB in the nervous system, we have investigated its expression in embryonic rat hippocampal pyramidal cell cultures and examined the effects of BDNF on signal transduction in the primary neurons. The expression of trkB transcripts was established by PCR analysis and in situ hybridization. In addition to gp145trkB, the pyramidal neuronal cultures expressed transcripts specific for the NT-3 receptor gp145trkC, but not for the high-affinity NGF receptor gp140trk or for p75LNGFR, a low-affinity receptor for all known members of the NGF family of neurotrophins including the gp145trkB ligands, BDNF and NT-4. The presence of gp145trkB receptors in the primary neuronal cultures was confirmed by immunocytochemical analysis in which > 90% of the cells stained with affinity purified polyclonal antibodies to gp145trkB. Immunoblots using this antibody revealed a single approximately 140 kDa protein in both adult hippocampus and pyramidal cultures. Addition of recombinant BDNF to these cultures induced the tyrosine phosphorylation of gp145trkB, as determined by antiphosphotyrosine staining of gp145trkB immunoprecipitates. Moreover, BDNF treatment activated the microtubule-associated protein (MAP) kinases, as determined by an increase in MAP2 phosphorylation in vitro. Both the 41 and 44 kDa forms of MAP kinase were activated by BDNF. BDNF also increased c-fos expression in over 90% of the cells. These results indicate that gp145trkB does not require p75LNGFR to form a functional receptor for BDNF in hippocampal pyramidal neurons. PMID- 8410188 TI - Purification and properties of m1-toxin, a specific antagonist of m1 muscarinic receptors. AB - The venom of the Eastern green mamba from Africa, Dendroaspis angusticeps, was found to block the binding of 3H-quinuclidinyl benzilate to pure m1 and m4 muscarinic ACh receptors expressed in Chinese hamster ovary cells. The principal toxin in the venom with anti-m1 muscarinic activity was purified by gel filtration and reversed-phase HPLC. This toxin has 64 amino acids, a molecular mass of 7361 Da, and an isoelectric point of 7.04. Its cysteine residues are homologous with those in curare-mimetic alpha-neurotoxins, and with those in fasciculin, which inhibits AChE. At low concentrations the toxin blocked m1 receptors fully and pseudoirreversibly while having no antagonist activity on m2 m5 receptors; the toxin is therefore named "m1-toxin." At higher concentrations m1-toxin interacted reversibly with m4 receptors, and half of the toxin dissociated in 20 min at 25 degrees C. The affinity of m1-toxin is therefore much higher for m1 than for m4 receptors. By comparison with m1-toxin, pirenzepine has sixfold higher affinity for m1 than for m4 receptors. Autoradiographs of muscarinic receptors in the rat brain demonstrated that m1-toxin blocked the binding of 2 nM 3H-pirenzepine only in regions known to bind m1-specific antibodies. Thus, m1-toxin is a much more selective ligand than pirenzepine for functional and binding studies of m1 muscarinic receptors. PMID- 8410189 TI - An electron microscopic analysis of hippocampal neurons developing in culture: early stages in the emergence of polarity. AB - In culture, hippocampal neurons initially establish several short, minor processes. The initial step in the emergence of polarity is marked by the rapid and selective growth of one of these processes, which becomes the axon. Subsequently the remaining processes become dendrites. We examined the ultrastructure of hippocampal neurons before and after the emergence of the axon. The minor processes in cells that had not yet formed axons were somewhat variable in appearance, but we found no ultrastructural feature that indicated which minor process might become the axon. The emergence of the axon was marked by several changes in its ultrastructure. The axon contained a sevenfold lower density of polyribosomes than the minor processes. In addition, axonal growth cones contained a pronounced concentration of membranous elements that resembled endoplasmic reticulum, elements that were rare in the growth cones of minor processes. Axons and minor processes did not differ in microtubule density. In order to gauge how rapidly these ultrastructural changes occur, we examined cells with short axons that, from their length, were estimated to have emerged only hours earlier. The preferential exclusion of polyribosomes from the axon and the concentration of reticular membrane in the axonal growth cone were already evident in such cells. These observations demonstrate that exclusion of ribosomes from the axon occurs early in development, about as soon as the axon can be identified. In contrast, previous work has shown that the differences in microtubule polarity orientation that distinguish mature axons and dendrites, and that have been proposed to account for the selective segregation of some constituents in neurons, first appear at a later stage of development (Baas et al., 1989). These observations also demonstrate that the accumulation of reticular membrane elements in growth cones, which has been noted previously, occurs preferentially in axonal growth cones and is closely correlated in time with the initial specification of the axon. The selective concentration of these elements in axonal growth cones could be associated with the uniquely rapid rate of axonal growth. PMID- 8410190 TI - Autoradiographic localization of serotonin receptor subtypes in cat visual cortex: transient regional, laminar, and columnar distributions during postnatal development. AB - Postnatal changes in the distribution of 5-HT receptor subtypes in the visual cortex of cats were assessed both qualitatively and quantitatively using in vitro autoradiographic methods. The 5-HT 1A, 1C, 2, and 3 receptor subtypes and the 5 HT uptake (5-HTUp) site were visualized with 3H-8-hydroxy-2(di-n-propyl amino)tetralin, 3H-mesulergine, (2,5-dimethoxy-4-125I-iodophenyl)-2-aminopropane, 3H-BRL43694, and 3H-cyanoimipramine, respectively. Although specific labeling of 5-HT3 receptors was not detected in the cat visual cortex at any age, each of the remaining 5-HT receptor subtypes exhibited unique temporal, regional, and laminar patterns of expression in visual cortical areas 17, 18, and 19 and lateral suprasylvian cortex (LS). 5-HT1A receptors were the earliest to demonstrate visual cortex-specific changes in expression. They exhibited peak levels of expression in all visual cortical areas, predominantly in supra- and infragranular layers, between postnatal day 10 (PD10) and PD30. Their levels in all areas declined progressively with increasing age. 5-HT1c receptors demonstrated their highest levels of expression in the deeper half of layer IV, but only in area 17, between PD40 and PD75. The most striking feature of their distribution throughout this period was that, in layer IV and extending into layer III, the 5-HT1c receptors were concentrated in columns that were 400 microns wide and had a center-to-center spacing of about 900 microns. This transient pattern of expression was not present beyond PD90. 5-HT2 receptors were most densely expressed in layer IV between PD40 and PD120, but they displayed a distinctly different distribution pattern. The densest binding of 5-HT2 receptors was limited to the upper half of layer IV and found in areas 17, 18, and LS. The distribution of layer 5-HT2 receptors along the dense band in layer IV of area 17 was discontinuous, exhibiting patches that were found in the same vertical columns as were the 5-HT1c receptors. Intermediate binding levels for the 5-HT2 receptors were found through layers I-III, the remainder of layer IV, and the subcortical white matter. The levels of 5-HT uptake sites increased gradually to reach adult levels by PD40, but with a distribution pattern that was basically homogeneous, both across cortical regions and across laminae.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410191 TI - Redistribution of cytoskeletal proteins in mammalian axons disconnected from their cell bodies. AB - Mice of the strain C57/BL/Ola exhibit a delay of Wallerian degeneration, such that axons survive for several weeks after a nerve transection that separates the axons from the cell bodies. In this Ola strain we have examined the distribution of cytoskeletal proteins in a 5 mm segment of the sciatic nerve for as long as 2 weeks after proximal and distal transections that prevent entry or exit of proteins via axonal transport. By 7 d after transections, there was a marked accumulation of alpha- and beta-tubulin, actin, and nonphosphorylated neurofilament epitopes at the proximal and at the distal ends of the transected axons, and loss of these proteins from the center of the isolated nerve segment. Highly phosphorylated neurofilament epitopes did not redistribute along the nerve, but there was a gradual loss of phosphorylated neurofilament immunoreactivity. These observations indicate the potential for bidirectional transport of a substantial portion of certain cytoskeletal proteins within axons of the PNS. PMID- 8410192 TI - Activation of protein kinase C by arachidonic acid selectively enhances the phosphorylation of GAP-43 in nerve terminal membranes. AB - Arachidonic acid (AA), a cis-unsaturated fatty acid that activates certain subspecies of protein kinase C (PKC), has been proposed to act as a retrograde messenger in modifying the efficacy of synapses during long-term potentiation (LTP). One prominent PKC substrate of the nerve terminal membrane, GAP-43 (F1, B 50, neuromodulin), shows an increase in phosphorylation that correlates with the persistence of LTP. The present study investigated whether AA might exert its effects on presynaptic endings by modulating the phosphorylation of GAP-43 and other membrane-bound proteins. Using synaptosomal membranes from the rat cerebrocortex, in which in vivo relationships between protein kinases and their native substrates are likely to be preserved, we found that in the absence of Ca2+, AA exerted a modest effect on the phosphorylation of GAP-43 and several other proteins; however, when AA was applied in conjunction with Ca2+, GAP-43 showed a particularly striking response: at Ca2+ levels likely to exist at the nerve terminal membrane during synaptic activity (10(-7) to 10(-5) M), AA (50 microM) increased the sensitivity of GAP-43 phosphorylation to Ca2+ by an order of magnitude, and increased its maximal level of phosphorylation by 50%. At resting Ca2+ levels, AA potentiated the stimulation in GAP-43 phosphorylation produced by 4 beta-phorbol 12,13-dibutyrate, a diacylglycerol (DAG) analog. The stimulatory effect of AA and its synergistic interaction with Ca2+ were found to be mediated by PKC, since they were blocked by a specific peptide inhibitor of PKC, [Ala25]PKC(19-31), but were unaffected by an inhibitor of protein phosphatase activity or by scavengers of free radicals. Since GAP-43 has been implicated in the development and plasticity of synaptic relationships, the synergistic effects of AA and the intracellular signals Ca2+ and DAG on the phosphorylation of GAP-43 may serve as an AND gate to modify presynaptic function and/or structure in response to coincident pre- and postsynaptic activity. PMID- 8410193 TI - Response of caudate neurons to stimulation of intrinsic and peripheral afferents in normal, symptomatic, and recovered MPTP-treated cats. AB - Cat caudate nucleus (CD) neurons were tested for changes in spontaneous activity, response to peripheral sensory stimuli (tactile, auditory, and visual), and electrical stimulation of monosynaptic afferents (pericruciate cortex and nucleus centralis lateralis) in normal cats and in the same cats after induction of and spontaneous recovery from parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). After normal baseline data were collected, cats were given MPTP (7.5 mg/kg, 5-7 d) to induce a parkinsonian syndrome consisting of rigidity, akinesia, and decreased orienting to sensory stimuli. During this symptomatic period, the mean spontaneous activity of CD units increased (6.20 spikes/sec vs 2.25 spikes/sec in normal cats). In these same animals, the percentage of units responding to peripheral sensory stimulation was significantly decreased (compared to normal) while the percentage of units responding to electrical stimulation of monosynaptic afferents increased. By 6 weeks after MPTP administration, cats had recovered gross motor and sensorimotor function and CD unit recordings were reinitiated. In functionally recovered animals, all electrophysiological measures returned to levels resembling those seen in normal animals. These data suggest that the processing of peripheral sensory information is an important part of basal ganglia function and that the sensory responsiveness of the CD may reflect the overall motor condition of the animal. The changes observed in the responsiveness of CD neurons to direct electrical stimulation of monosynaptic afferents may indicate that the defect in the processing of polysynaptic sensory information observed in the striatum in parkinsonian animals may be occurring, at least in part, extrastriatally. PMID- 8410194 TI - The mechanism of tonic inhibition of crayfish escape behavior: distal inhibition and its functional significance. AB - The excitability of crayfish escape behavior is seldom fully predictable. A major determinant of this fickleness is a form of descending inhibition that is reliably evoked during restraint or feeding and is called "tonic inhibition." Tonic inhibition was found to inhibit postsynaptically the lateral giant neurons, the command neurons for one form of escape. This inhibition is located on lateral giant dendrites that are electrotonically distant from the neuron's spike initiating zone. in contrast, the postsynaptic inhibition due to "recurrent inhibition," which prevents new escape responses from starting while a previously initiated one is in process, occurs proximally, near the spike initiating zone. The distalness of tonic inhibition could be an adaptation for selective suppression of parts of the lateral giant dendritic tree. Consistent with this, evidence was obtained that the tonic inhibitory system can suppress responses to specific sensory fields. An independent reason for targeting recurrent inhibition proximally and tonic inhibition distally was suggested by the functional requirements of each inhibitory process: recurrent inhibition needs to be "absolute" in the sense that the response should be absolutely prevented, whereas it must be possible to override tonic inhibition. Neuronal models demonstrated that proximal inhibition gives recurrent inhibition the required property of absoluteness while distal inhibition allows tonic inhibition to be overridden ("relativity"). It was shown that the relativity of distal inhibition arises from its interaction with the process of saturation of excitation and that tonic inhibition does indeed interact with excitatory saturation as predicted. It is suggested that the property of relativity of distal inhibition is exploited in other nervous systems as well. PMID- 8410195 TI - Crayfish tonic inhibition: prolonged modulation of behavioral excitability by classical GABAergic inhibition. AB - Previous studies have indirectly implicated the two neurotransmitters 5-HT and GABA in mediating tonic inhibition of the crayfish lateral giant (LG) escape reaction. In this study, pharmacological agents were selectively delivered to restricted portions of the abdominal CNS (where LG escape circuitry resides) to assess directly the role of these two transmitters in tonic inhibition. Both 5-HT and GABA depressed monosynaptic, electrical transmission to the LG neurons, the command neurons for LG escape, and application of either transmitter resulted in a depolarizing conductance increase in the LG neuron. The effects of 5-HT persisted in preparations in which chemical transmission was effectively abolished, implying that there are 5-HT receptors on the LG neuron itself, along with the known GABA receptors. Restricted delivery of the GABA chloride channel blocker picrotoxin to only the abdominal CNS blocked the expression of tonic inhibition there (without interfering with the rostral generation of tonic inhibition). Therefore, if 5-HT mediated tonic inhibition, the effects of 5-HT on the abdomen should also be antagonized by picrotoxin. However, this was not the case, thus suggesting that 5-HT does not mediate tonic inhibition. The most likely neurotransmitter used for tonic inhibition is GABA acting via ligand-gated chloride channels. Thus, although this form of behavioral modulation can be tonically active for very long periods, it nevertheless appears to be mediated by a classical synaptic mechanism. PMID- 8410196 TI - Membrane properties of ameboid microglial cells in the corpus callosum slice from early postnatal mice. AB - Microglial cells in culture are distinct from neurons, macroglial cells, and macrophages of tissues other than brain with respect to their membrane current pattern. To assess these cells in the intact tissue, we have applied the patch clamp technique to study membrane currents in microglial cells from acute, whole brain slices of 6-9-d-old mice in an area of microglial cell invasion, the cingulum. As strategies to identify microglial cells prior to or after recording, we used binding and incorporation of Dil-acetylated low-density lipoproteins, binding of fluorescein isothiocyanate-coupled IgG via microglial Fc-receptors, and ultrastructural characterization. As observed previously for cultured microglial cells, depolarizing voltage steps activate only minute if any membrane currents, while hyperpolarizing voltage steps induced large inward currents. These currents exhibited properties of the inwardly rectifying K+ channel in that the reversal potential depended on the transmembrane K+ gradient, inactivation time constants decreased with hyperpolarization, and the current was blocked by tetraethylammonium (50 mM). This study represents the first attempt to assess microglial cells in situ using electrophysiological methods. It opens the possibility to address questions related to the function of microglial cells in the intact CNS. PMID- 8410197 TI - Seizures and brain injury in neonatal rats induced by 1S,3R-ACPD, a metabotropic glutamate receptor agonist. AB - The role of metabotropic excitatory amino acid receptors in seizures and brain injury was examined using the selective metabotropic agonist 1S,3R-ACPD [(1S,3R) 1-aminocyclopentane-1-3-dicarboxylic acid] in 7-d-old neonatal rats. Systemic administration of 1S,3R-ACPD produced dose-dependent convulsions (ED50 = 16 mg/kg, i.p.) that were stereoselective for the active metabotropic ACPD isomer, since 1R,3S-ACPD was less potent (ED50 = 93 mg/kg, i.p.). 1S,3R-ACPD-induced seizures were antagonized by systemic administration of dantrolene, an inhibitor of intracellular calcium mobilization, but not by the ionotropic glutamate antagonists MK-801 or GYKI-52466. As indexed by hemispheric brain weight differences 5 d postinjection, unilateral intrastriatal injection of 1S,3R-ACPD (0.1-2.0 mumol/microliters), but not 1R,3S-ACPD, produced dose-dependent brain injury (maximal effect of 3.4 +/- 0.5% damage). 1S,3R-ACPD brain injury occurred in the absence of prominent behavioral convulsions. Histologic and ultrastructural examination of 1S,3R-ACPD-injected rat brains revealed swelling and degeneration of select neurons at 4 hr postinjection, but little evidence of injured neurons 5 d later. 1S,3R-ACPD-mediated brain injury was not attenuated by systemic administration of the NMDA antagonist MK-801 or the AMPA antagonist GYKI 52466. However, cointrastriatal injection of dantrolene reduced the severity of 1S,3R-ACPD injury by 88 +/- 7%. These studies indicate that seizures and neuronal injury can be elicited by the selective activation of metabotropic glutamate receptors in perinatal rats, and these effects of 1S,3R-ACPD involve the mobilization of intracellular calcium stores. PMID- 8410198 TI - Two distinct rhythmic motor patterns are driven by common premotor and motor neurons in a simple vertebrate spinal cord. AB - Xenopus embryos show two distinct rhythmic motor patterns: swimming and struggling. Both can be generated by spinal cord circuitry and evoked by stimulation of a single skin sensory pathway (Soffe, 1991b). This presents a valuable opportunity to explore mechanisms for vertebrate motor pattern switching. Swimming and struggling have been compared using intracellular recording from spinal neurons in immobilized embryos. Underlying synaptic drive was similar; motoneurons and premotor interneurons were excited in phase with ipsilateral motor root discharge and inhibited in phase with contralateral motor root discharge. Excitation was stronger during struggling and associated with short bursts of impulses, contrasting with single spikes per cycle during swimming. Excitation was reduced in both patterns by local application of 1 mM kynurenic acid, indicating excitatory amino acid mediation. Inhibition was antagonized by 1 microM strychnine, indicating glycine mediation. Many motoneurons (76%) and premotor interneurons (68%) fired during both swimming and struggling, including examples of all three spinal premotor interneuron classes. Most of the remaining motoneurons (20%) and premotor interneurons (24%) fired only during struggling, providing roughly 30% more active neurons than during swimming. To investigate whether new neuronal classes become active during struggling, recordings were made from sensory neurons and sensory interneurons. Rohon-Beard sensory neurons did not fire during either swimming or struggling. Dorsolateral commissural sensory interneurons received rhythmic, strychnine sensitive inhibition during both swimming and struggling and also did not fire. Neither of these neuronal classes is therefore recruited to the circuitry for struggling. Although behaviorally distinct, Xenopus embryo swimming and struggling motor patterns appear to employ similar synaptic drive. I propose that this reflects the common nature of much of the premotor circuitry that drives them. Extra neurons are recruited to this circuitry during struggling, but only from within classes that also participate in swimming. PMID- 8410199 TI - Loss of glutamate decarboxylase mRNA-containing neurons in the rat dentate gyrus following pilocarpine-induced seizures. AB - In situ hybridization methods were used to determine if glutamic acid decarboxylase (GAD) mRNA-containing neurons within the hilus of the dentate gyrus are vulnerable to seizure-induced damage in a model of chronic seizures. Sprague Dawley rats were injected intraperitoneally with pilocarpine, and the hippocampal formation was studied histologically at 1, 2, 4, and 8 week intervals after pilocarpine-induced seizures. In situ hybridization histochemistry, using a digoxigenin-labeled GAD cRNA probe, demonstrated a substantial decrease in the number of GAD mRNA-containing neurons in the hilus of the dentate gyrus in the pilocarpine-treated rats as compared to controls at all time intervals. Additional neuronanatomical studies, including cresyl violet staining, neuronal degeneration methods, and histochemical localization of glial fibrillary acidic protein, suggested that the decrease in the number of GAD mRNA-containing neurons was related to neuronal loss rather than to a decrease in GAD mRNA levels. The loss of GAD mRNA-containing neurons in the hilus contrasted with the relative preservation of labeled putative basket cells along the inner margin of the granule cell layer. Quantitative analyses of labeled neurons in three regions of the dentate gyrus in the 1 and 2 week groups showed statistically significant decreases in the mean number of GAD mRNA-containing neurons in the hilus of both groups of experimental animals. No significant differences were found in the molecular layer or the granule cell layer, which included labeled neurons along the lower margin of the granule cell layer. The results indicate that, in this model, a subpopulation of GAD mRNA-containing neurons within the dentate gyrus is selectively vulnerable to seizure-induced damage. Such differential vulnerability appears to be another indication of the heterogeneity of GABA neurons. PMID- 8410200 TI - Structural and functional specialization of A delta and C fiber free nerve endings innervating rabbit corneal epithelium. AB - An in vitro preparation of rabbit cornea was used to compare anatomical specialization and electrophysiological responses of A delta and C fiber sensory afferents which terminate as free nerve endings. Living nerve endings were visualized using epifluorescence microscopy and the vital dye 4-di2-ASP, and response properties were determined using microstimulation and recording of fiber discharge activity. Fiber type was determined based on conduction velocity measurement and preferred stimulus energy (modality) of each fiber. Four modality specific fiber populations were identified: (1) slowly adapting C fiber cold receptors (conduction velocity of 0.25-1.6 m/sec), (2) C fiber chemosensitive units with mixed phasic and tonic activity (1.1-1.8 m/sec), (3) rapidly adapting mechanosensitive A delta fibers (1.5-2.8 m/sec), and (4) high-threshold mechano/heat (> 350 dyne or > 40 degrees C) phasic A delta afferents (3.5-4.4 m/sec). In addition to these physiological differences, anatomical specialization was also noted. A delta fiber nerve endings were distinguished from those of C fibers by thin, elongated sensory endings that ran parallel to the corneal surface; C fiber endings formed short, branching clusters that ran mostly perpendicular to the surface. The elongated structure of A delta nerve endings was associated with directional selectivity for mechanical stimuli. These results substantiate previous suggestions that free nerve endings can exhibit both structural and functional specialization. PMID- 8410201 TI - Reduced levels of acetylcholine receptor expression in chick ciliary ganglion neurons developing in the absence of innervation. AB - Chick ciliary ganglion neurons receive innervation from a single source, the accessory oculomotor nucleus (AON), and nicotinic ACh receptors (AChRs) mediate chemical synaptic transmission through the ganglion. Previous experiments examining the developmental expression of AChRs in embryonic chick ciliary ganglion neurons in situ have shown that AChR levels increase substantially in the neurons at the time of innervation. Prior to synapse formation, few AChRs are detected in the neurons. In the present experiments, the role of presynaptic inputs in inducing an increase in AChRs was established by examining AChR levels in ciliary ganglion neurons that have been deprived of innervation by surgical ablation of the AON prior to synapse formation. AChR levels were dramatically reduced in neurons of input-deprived ganglia as compared to control innervated neurons at all developmental stages examined from embryonic day (ED) 5 to ED 12 as determined by indirect immunocytochemical labeling of frozen ganglion sections with the anti-AChR monoclonal antibody mAb 35, and light microscopy. In contrast, neuronal somata of input-deprived and control ganglia had equivalent levels of immunolabeling for three other components, a transmembrane glycoprotein of synaptic vesicles, SV2, and two microtubule-associated proteins, MAP 1B and MAP 2, from ED 5 up to ED 10. The results demonstrate that presynaptic inputs specifically increase the levels of AChR expression in developing neurons. In addition, changes in the levels of immunolabeling for AChRs, SV2, MAP 1B, and MAP 2 in neuronal somata after ED 10 demonstrate that other major developmental events also influence the levels of these components in neurons. Declines in the intensity of AChR, SV2, MAP 1B, and MAP 2 immunolabeling within a subset of neuronal somata in both operated and control ganglia at ED 10 and 12 coincide with the period of neuronal cell death. Increases in AChR labeling in the rest of the neuronal population of input-deprived ganglia at ED 12 suggest that, in addition to innervation, synapse formation with the peripheral target tissue influences AChR levels in developing neurons in situ. PMID- 8410202 TI - Auditory compensation for early blindness in cat cerebral cortex. AB - Single-neuron activity was recorded in the caudal part of the anterior ectosylvian (AE) cortex of cats that had been deprived of vision for several years by means of binocular lid suture shortly after birth and in normal control animals. Over 300 neurons were tested in each group with auditory, visual, and somatosensory stimuli. We confirmed the existence of an anterior ectosylvian visual area (AEV) in the fundus and ventral bank of the AE sulcus. Neurons in AEV had purely visual responses in normal cats. In visually deprived cats, by contrast, only a minority of cells in this area still responded to visual stimulation. Instead, most cells reacted vigorously to auditory and, to some extent, somatosensory stimuli. The few remaining visual neurons were also driven by auditory or somatosensory stimuli. No increase in the number of unresponsive neurons was found. It appears, therefore, that a cortical region that normally represents visual activity can become driven by auditory or somatosensory activity as a result of visual deprivation. Our results imply that early blindness causes compensatory increases in the amount of auditory cortical representation, possibly by an expansion of nonvisual areas into previously visual territory. In particular, they provide evidence for the existence of neural mechanisms for intermodal compensatory plasticity in the cerebral cortex of young animals. The changes described here may also provide the neural basis for a behavioral compensation for early blindness described elsewhere. PMID- 8410203 TI - Neural substrates of visual stimulus-stimulus association in rhesus monkeys. AB - Rhesus monkeys learned 10 visual stimulus-stimulus association, or paired associates. They then received bilateral removals of either the amygdaloid complex and underlying cortex, the hippocampal formation and underlying cortex, or both combined, or they were retained as unoperated controls. After surgery or rest, the monkeys were tested for their retention of the preoperatively learned set of paired associates, as well as for their ability to learn new associations of the same type. Both unoperated controls and hippocampectomized monkeys relearned the preoperatively trained set of paired associates almost immediately. By contrast, monkeys with amygdala removals were moderately retarded in relearning, and monkeys with combined amygdala and hippocampal ablations were severely retarded. When confronted with new sets of visual stimuli, monkeys with amygdala removals or hippocampal removals learned new sets of paired associates at the same rate as the controls, whereas monkeys with the combined ablation were again profoundly retarded. Only one monkey with the combined lesion was able to learn new stimulus-stimulus associations to criterion, and then only after extensive training, despite the ability of all three animals in this group to perform delayed matching-to-sample with the same stimuli and the same intraatrial delays as those used in the paired associate task. At the end of the main experiment, two of the unoperated controls received bilateral ablations of the rhinal cortex. These monkeys showed the same level of difficulty in learning new paired associates as the animals in the main experiment that had received the combined amygdala plus hippocampal ablations. The results implicate the medial temporal lobe, and particularly the rhinal cortex, in the formation of stimulus stimulus associative memories. PMID- 8410204 TI - Collagenase and collagenase inhibitors secreted by cultured human periodontal ligament cells. AB - We have developed a new assay method for collagenase activity, in the presence of two different types of collagenase inhibitors, in culture medium of periodontal ligament (PDL) cells. Human PDL cells were cultured with serum-free alpha-MEM for 5 days after reaching confluency, and the culture medium was then harvested. Collagenase and collagenase inhibitors in the medium were concentrated by ammonium sulfate precipitation. The precipitate was dissolved, and the suspension was applied to an Ultrogel AcA 54 column for partial purification. Collagenase inhibitor activity was detected as two peaks; a small inhibitor molecule of about 32 kDa sensitive only to dithiothreitol (DTT), and a large one of about 70 kDa considerably more sensitive to 4-aminophenylmercuric acetate (APMA) than to DTT. Therefore, pretreatment of the sample preparation with both DTT and APMA was necessary in order to inactivate the inhibitors prior to assay of latent collagenase. This observation indicates that PDL cells secrete collagenase and two collagenase inhibitors without any stimulation in vitro. PMID- 8410205 TI - Finite element model of facial soft tissue. Effects of thickness and stiffness on changes following simulation of orthognathic surgery. AB - A study was conducted to investigate the influence of the thickness and stiffness of soft tissue on changes following orthognathic surgery. The finite element method was used to simulate surgery. Three finite element models with normal thickness and stiffness, twice the thickness and twice the stiffness were made in preparation for the simulation. The thickness of soft tissue had more effect than stiffness on posterior and vertical changes. PMID- 8410206 TI - Amelanotic melanoma of the oral cavity. AB - A case of amelanotic melanoma arising in the upper molar region, which was difficult to diagnose histologically, is reported. The patient was a 79-year-old woman, who complained of a painful swelling in the gingiva of the left upper molar region. Routine histological examination showed that the lesion was composed of diffusely scattered atypical cells with round, spindle-shaped and irregular nuclei and scanty fibrous connective tissue. A fascicular arrangement was often found in the lesion, and no cancer nests were observed. Immunohistochemical study demonstrated positive staining for S-100 protein in both the nuclei and cytoplasm of the tumor cells. Electron microscopic examination revealed that cell organelles were abundant, and an interrupted basal lamina was often found along the cell membrane. The preliminary diagnosis was a non-epithelial malignant tumor. After surgery, histological examination of metastases in lymph nodes from the submandibular region revealed that the tumor cells contained melanin pigment in the cytoplasm, as confirmed by Masson's melanin stain. The final pathological diagnosis was therefore amelanotic melanoma. Immunohistochemical staining for S-100 protein may be useful for differential diagnosis of amelanotic melanoma in conjunction with electron microscopic examination. PMID- 8410207 TI - Effect of an anticalculus toothpaste containing pyrophosphate on pyrophosphatase activity and the state of calcium phosphate in saliva. AB - Experiments were performed to determine if a pyrophosphate-containing anticalculus toothpaste could affect certain salivary constituents involved in the mineralization of dental plaque. The findings indicated no obvious changes in acid and alkaline pyrophosphatases, ionized calcium, total calcium, inorganic phosphate or pH in saliva for 1-135 min after brushing the teeth with the anticalculus toothpaste. Data obtained by brushing with the toothpaste three times daily for two weeks also confirmed these results. Our findings clearly indicate that pyrophosphate-stabilizing agents in the anticalculus toothpaste are not fully effective in the oral cavity. In addition, the pyrophosphate-containing toothpaste has no influence on the state of calcium and phosphate in saliva. PMID- 8410208 TI - Neutrophil chemotaxis and periodontal status in Down's syndrome patients. AB - Along with clinical parameters, chemotaxis and random migration of neutrophils were evaluated in 15 patients with Down's syndrome (DS) and 15 healthy subjects. Signs of more severe gingival inflammation were present in the DS group. The random migration and chemotaxis of neutrophils were significantly decreased in comparison with the control group. In DS, the pathological status was attributed to these impaired host defense factors besides the existing bacterial plaque. PMID- 8410209 TI - Allogeneic transplants of rib cartilage preserved in 98% glycerol or 70% alcohol into the malar process of rats: a comparative histological study. AB - A comparative study was made of two methods of cartilage preservation, 98% glycerol and 70% alcohol. Rib cartilage was treated by either of these methods and transplanted into the malar process of rats. Cartilage grafts preserved by both methods were equally well tolerated. Resorption and bone substitution were similar in both groups after 120 days, although resorption was greater for the alcohol-preserved cartilage up until day 30. The possible reduction in antigenicity by the 98% glycerol did not produce any difference of behavior from the cartilage preserved in 70% alcohol. PMID- 8410210 TI - Don't read this! You won't like it! But it is nevertheless true. PMID- 8410211 TI - Sinus lift as a means of improving restorative options in the edentulous maxilla: a case report. PMID- 8410212 TI - Healing the children--a Guatemalan experience. PMID- 8410213 TI - Periodontal screening & recording an early detection system. PMID- 8410214 TI - Increased stroke risk predicted by compromised cerebral blood flow reactivity. AB - The authors sought to determine risk for stroke in individuals with symptomatic carotid stenosis or occlusion based upon an assessment of cerebral blood flow (CBF) reserves. Vascular reserve was assessed by two consecutive xenon/computerized tomography (Xe/CT) CBF studies with intravenous acetazolamide introduced 20 minutes prior to the second study. Patients were assigned to one of two vasoreactivity groups. Group 2 included individuals who experienced a CBF reduction of more than 5% in at least one vascular territory and had a baseline flow of 45 cc/100 gm/min or less. Group 1 included all other individuals. Any territory with volume loss on CT of more than 50% was eliminated from analysis. Sixty-eight individuals were followed at 6-month intervals for a mean of 24 months. In Group 1 two strokes were observed contralateral to the side with lowest reserve, for a stroke incidence of 4.4%; in Group 2 eight strokes were observed ipsilateral to the side with lowest reserve, for a stroke incidence of 36%. The latter group had a 12.6 times greater chance of stroke (p = 0.0007). History of stroke, history of transient ischemic attacks, baseline CBF, and degree of stenosis were not associated with an increased stroke rate. In this study, significantly compromised vascular reserves accompanied by relatively low initial flow identified individuals who subsequently demonstrated a significantly increased rate of ipsilateral stroke. PMID- 8410215 TI - Abnormal cerebral vasodilation in aneurysmal subarachnoid hemorrhage: use of serial 133Xe cerebral blood flow measurement plus acetazolamide to assess cerebral vasospasm. AB - A patient with cerebral vasospasm following subarachnoid hemorrhage (SAH) was investigated by serial measurement of cerebral blood flow (CBF) using the xenon 133 emission tomography method. The CBF was measured before and after acetazolamide injection. On Day 2 after SAH, there was early local hyperperfusion in the middle cerebral artery (MCA) territory, ipsilateral to the left posterior communicating artery aneurysm. The regional CBF of this arterial territory decreased slightly after acetazolamide injection, probably because of vasoplegia and the "steal" phenomenon, and thus surgery was delayed. A right hemiplegia with aphasia and disturbed consciousness occurred 4 days later (on Day 6 after SAH) due to arterial vasospasm, despite treatment with a calcium-channel blocker. The initial hyperemia of the left MCA territory was followed by ischemia. The vasodilation induced by acetazolamide administration was significantly subnormal until Day 13, at which time CBF and vasoreactivity amplitude returned to normal and the patient's clinical condition improved. Surgery on Day 14 and outcome were without complication. It is concluded that serial CBF measurements plus acetazolamide injection are useful for monitoring the development of cerebral vasospasm to determine the most appropriate time for aneurysm surgery. PMID- 8410216 TI - Cardiac performance enhancement from dobutamine in patients refractory to hypervolemic therapy for cerebral vasospasm. AB - The use of the beta-agonist dobutamine in combination with hypervolemic preload enhancement of cardiac performance was analyzed in 23 patients who failed to respond to traditional preload enhancement following aneurysmal subarachnoid hemorrhage. The patients ranged in age from 13 to 82 years, and three had a history of cardiac disease. Each patient underwent placement of a flow-directed balloon-tipped catheter and the following measurements were obtained during hyperdynamic therapy: pulmonary artery wedge pressure, central venous pressure, cardiac index, stroke volume index, total peripheral resistance, and left ventricular stroke work index (LVSWI). Mean baseline cardiac function was found to be within normal limits (LVSWI = 47.6 +/- 4.2 gm/min/sq m and cardiac index = 3.30 +/- 0.22 liter/min/sq m). After baseline measurements were recorded, 5% albumin was infused at 300 cc/hr and dobutamine was initiated at a rate of 5 to 10 micrograms/kg/hr. This hyperdynamic therapy with dobutamine in the presence of volume loading resulted in a 52% increase in cardiac index, a 15% increase in LVSWI, and a 21% decrease in total peripheral resistance. The clinical reversal of ischemic symptoms due to subarachnoid hemorrhage was evident in 18 (78%) of the 23 patients. PMID- 8410217 TI - Effects of timing of methylprednisolone or naloxone administration on recovery of segmental and long-tract neurological function in NASCIS 2. AB - Previous analyses of the National Acute Spinal Cord Injury Study (NASCIS) have not distinguished recovery of segmental function at the injury level from recovery of the long spinal tracts. Recovery at the injury level could be of considerable clinical significance, but long-tract recovery is the ultimate therapeutic goal. This analysis demonstrates that the greatest proportion of all neurological recovery and of recovery due to treatment with very high doses of methylprednisolone within 8 hours of injury occurs below the lesion. Methylprednisolone treatment administered early following injury has been found to improve recovery below the lesion in patients initially diagnosed as having complete or incomplete injuries; it also leads to greater (but still relatively small) improvement in the injury level. The analysis indicates that delayed treatment with methylprednisolone is associated with decreased neurological recovery. Naloxone administration also improved neurological function below the lesion in patients with incomplete injuries; these results support further experimental work with this drug. This observation of differential neurological response within a narrow treatment window has important implications for both experimental studies and clinical management. Early clinical management with high dose methylprednisolone is supported by this analysis. PMID- 8410218 TI - The petrosal approach with hearing preservation. AB - Twenty-six patients with petroclival lesions were operated on via a petrosal approach designed to preserve hearing. The surgical pathology included 14 meningiomas, three chordomas, three epidermoid cysts, four vertebrobasilar aneurysms, and two pontine cavernous malformations. The approach allowed complete resection of 14 of 20 tumors and definitive treatment of all six vascular lesions. Complications included cerebrospinal fluid leakage in three patients, high-frequency sensorineural hearing loss in three, meningitis in one, and cranial nerve palsies (which were usually transient). This approach allows a wide exposure of the petroclival region with decreased operating distance. Cerebellar and temporal lobe retraction are minimized, dural sinus patency is maintained, and the inner ear structures are not sacrificed. The approach is suitable for neoplastic or vascular lesions involving the petroclival region, the ventral pons, or the basilar artery trunk. The surgical technique, indications, and neuro otological considerations are discussed. PMID- 8410219 TI - Intracanalicular acoustic neuroma: early surgery for preservation of hearing. AB - Diagnosis of very small acoustic neuromas has become much more common since the advent of magnetic resonance imaging. Many of the patients so diagnosed have minimal unilateral hearing loss as their only symptom. Because limited information is available on the natural history and prognosis of these lesions, the choice of treatment is controversial. The authors review their recent experience with the surgical treatment of intracanalicular acoustic neuroma. The records of 14 consecutive patients with intracanalicular acoustic neuroma were reviewed with respect to type of presentation, pre- and postoperative facial and auditory nerve function, surgical approach, and complications. Detailed results for patients operated on to preserve hearing are presented. Presenting symptoms were nearly equally divided among diminished hearing, vertigo, and tinnitus. Eleven of the 14 patients had serviceable hearing preoperatively and nine (82%) remained in this condition postoperatively. Facial nerve function was unchanged by operation in 12 patients. Seven operations were performed through the middle fossa, five through the posterior fossa, and two by the translabyrinthine approach. The probability of preserving hearing during surgical excision of intracanalicular acoustic neuroma in patients with serviceable hearing exceeds 80%. Given the relative infrequency of serious complications and the likelihood of progressive hearing loss in the untreated patient, excision of such small tumors shortly after diagnosis may offer the best chance of long-term hearing preservation. PMID- 8410220 TI - Thyrotropin-producing pituitary adenomas. AB - To evaluate the biology of thyrotropin (TSH)-producing pituitary adenomas, the authors reviewed the charts of 19 patients who underwent transsphenoidal surgery within a 15-year period at the University of California, San Francisco (UCSF). Between 1989 and 1991, the period during which immunostaining techniques were used consistently for diagnosis, 2.8% of the pituitary adenomas treated at UCSF were TSH-producing. The rate of reoperation for tumor recurrence was 10.5%. Before pituitary surgery, more than one-third of the 19 patients had undergone thyroid ablation. Two patients had a history of Hashimoto's thyroiditis. The female:male ratio was 1.7:1. Women tended to develop these tumors at a younger age and had a longer history of symptoms but their tumors were smaller and less often invasive than those seen in men. About 50% of the tumors were purely TSH producing and 50% were plurihormonal, including five that produced both TSH and adrenocorticotroph hormone. All tumors were macroadenomas. Before surgery, 46% of the patients had abnormal electrocardiographic findings; 16% had a rapid onset of severe neurological conditions either before or after surgery. It is concluded that TSH-producing adenomas are more common in patients who undergo surgical treatment than was previously thought. In addition, they occur more frequently in women, have a different biology in women than in men, and tend to be associated with potentially life-threatening cardiovascular and neurological complications. PMID- 8410221 TI - Spinal malignant nerve-sheath tumor or cellular schwannoma? A striking difference in prognosis. AB - Cellular schwannoma, a recently delineated entity, has a histological appearance mimicking that of malignant neoplasms. The aim of this study was to determine the outcome for patients treated for a spinal cellular schwannoma or malignant nerve sheath tumor. A histological re-examination was conducted of 283 spinal tumors, considered to originate from a nerve root, that were treated in the Department of Neurosurgery between 1953 and 1985. After re-examination, 50 of these were determined to be other tumors or non-neoplastic lesions. The review yielded eight cellular schwannomas and six malignant nerve-sheath tumors out of 233 of nerve sheath origin. Immunohistochemical staining with a commercially available polyclonal antibody against S-100 protein was positive in all cases of cellular schwannoma, but negative for the malignant tumors. Clinical outcome was favorable for patients with cellular schwannomas, but uniformly poor for those with the malignant tumors. PMID- 8410222 TI - Unreliability of contemporary neurodiagnostic imaging in evaluating suspected adult supratentorial (low-grade) astrocytoma. AB - Many physicians rely upon neuroimaging studies alone to select therapy for adult patients suspected of having a glial neoplasm, in the belief that certain imaging features accurately characterize the histological diagnosis of low-grade astrocytoma. During a 4-year interval when both computerized tomography and magnetic resonance imaging was available, the authors performed stereotactic biopsies on 20 consecutive adult patients who were suspected of having an astrocytoma. The patients were generally young (mean age 37 years), had seizures (17 cases), and had lobar lesions. An accurate histological diagnosis was obtained, without morbidity, in all 20 patients. Only 10 (50%) in fact had low grade astrocytomas, whereas nine (45%) had anaplastic astrocytomas and one (5%) had encephalitis. The results of this study indicate that modern high-resolution neuroimaging alone cannot be used as a reliable tool to predict the histological diagnosis of astrocytoma (50% false-positive rate). All patients with supratentorial mass lesions that exhibit the "typical" imaging features of astrocytoma should undergo stereotactic biopsy for confirmation in order that appropriate management may be planned. PMID- 8410223 TI - Calpain-calpastatin system of canine basilar artery in vasospasm. AB - Vasospasm was produced in the canine basilar artery by a two-hemorrhage method, while contraction was induced in the normal canine basilar artery by a local application of KCl or serotonin after transclival exposure. The control animals were injected with saline instead of fresh blood. The activation of mu-calpain, a Ca(++)-dependent neutral protease, in the basilar artery was studied by evaluating the conversion from its inactivated into its activated form on immunoblots. In addition, the activity of calpastatin, an intrinsic inhibitor of calpain, in the basilar artery was determined by assay. The majority of the mu calpain was inactivated in the control group. In the spastic group, mu-calpain was generally activated markedly in the early stage of vasospasm and moderately thereafter. The contraction induced by KCl or serotonin application was classified into the early phasic and the later tonic stages; mu-calpain was usually activated in the phasic stage and inactivated in the tonic stage. Calpastatin activity was significantly decreased during vasospasm, whereas it was not significantly changed in KCl- or serotonin-induced contraction. The final activity of mu-calpain results from the balance of mu-calpain and calpastatin. This suggests that mu-calpain activity was enhanced continuously in the spastic group and transiently in the KCl or serotonin group, and that the continuous activation of mu-calpain during vasospasm probably induced more proteolytic changes compared to those in the KCl or serotonin group. PMID- 8410224 TI - Intrathecal baclofen down-regulates GABAB receptors in the rat substantia gelatinosa. AB - At present, it is not clear why drug tolerance develops in patients receiving intrathecal baclofen for the chronic treatment of spasticity of spinal origin. To investigate the mechanisms of tolerance to the gamma-aminobutyric acid (GABAB) agonist baclofen, rats were implanted with intrathecal catheters and continuously infused with either the drug or saline for 1, 3, or 7 days. The dose chosen, 1 microgram/hr, initially caused profound hindlimb motor weakness, but by Day 7 the rats had adapted to the drug and exhibited only minimal motor impairment. The animals were sacrificed on Day 1, 3, or 7 and quantitative autoradiography was used to determine the binding density and affinity of the GABAB receptors in the substantia gelatinosa of the lumbar spinal cord. After 1 day of drug infusion there was no change in binding parameters, but after 3 and especially after 7 days there was a significant decrease in the GABAB binding density (74% and 66%, respectively) in baclofen-treated rats as compared to saline-treated control rats. This GABAB receptor down-regulation correlated with tolerance to the motor weakness in the baclofen-treated animals and suggests that similar mechanisms contribute to drug tolerance in patients. PMID- 8410225 TI - Experimental chronic compressive cervical myelopathy. AB - A canine model simulating both cervical spondylosis and its results in delayed progressive myelopathy is presented. This model allowed control of compression, an ongoing assessment of neurological deficits, and evaluation using diagnostic images, frequent electrophysiological tests, local blood flow measurements, and postmortem histological examinations. Subclinical cervical cord compression was achieved in 14 dogs by placing a Teflon washer posteriorly and a Teflon screw anteriorly, producing an average of 29% stenosis of the spinal canal. Four dogs undergoing sham operations were designated as controls. Twelve of the animals undergoing compression developed delayed and progressive clinical signs of myelopathy, with a mean latent period to onset of myelopathy of 7 months. Spinal cord blood flow studies using the hydrogen clearance method showed a significant transient increase in blood flow immediately after compression and a decrease before sacrifice. Somatosensory evoked potential studies indicated progressive deterioration during the period of compression. Magnetic resonance images revealed intramedullary changes. Histological studies showed abnormalities overwhelmingly within the gray matter, including changes in vascular morphology, loss of large motor neurons, necrosis, and cavitation. Axonal degeneration and obvious demyelination were rarely seen. The most profound morphological changes occurred at the site of greatest compression. It is proposed that a momentary arrest of microcirculation occurs during extension of the neck because of loss of the reserve space in the compromised spinal canal. This microcirculatory disturbance is predominant in the watershed area of the cord and mainly affects the highly vulnerable anterior horn cells, leading to neuronal death, necrosis, and eventual cavitation at the junction of the dorsal and anterior horns. Additional supportive evidence of this hypothesis was derived from the literature. PMID- 8410226 TI - The role of photodynamic therapy in posterior fossa brain tumors. A preclinical study in a canine glioma model. AB - Photodynamic therapy was studied in dogs with and without posterior fossa glioblastomas. This mode of therapy consisted of intravenous administration of Photofrin-II at doses ranging from 0.75 to 4 mg/kg 24 hours prior to laser light irradiation in the posterior fossa. Tissue levels of Photofrin-II were four times greater in the tumor than in the surrounding normal brain. Irradiation was performed using 1 hour of 500 mW laser light at a wavelength of 630 nm delivered through a fiberoptic catheter directly into the tumor bed via a burr hole. All animals receiving a high dose (4 or 2 mg/kg) of Photofrin-II developed serious brain-stem neurotoxicity resulting in death or significant residual neurological deficits. A lower dose (0.75 mg/kg) of Photofrin-II produced tumor kill without significant permanent brain-stem toxicity in either the control animals or the animals with cerebellar brain tumors receiving photodynamic therapy. PMID- 8410227 TI - Effect of pentobarbital on normal brain protection and on the response of 9L rat brain tumor to radiation therapy. AB - The authors attempted to confirm published reports that pentobarbital protects against radiation-induced damage to normal rat brain, as well as enhances radiotherapeutic efficacy in a rat brain tumor model. They evaluated animal survival in 9L gliosarcoma-burdened rats that received whole-brain radiation therapy (16, 24, 32, or 40 Gy) while under intraperitoneal pentobarbital (60 mg/kg) or intramuscular ketamine (60 mg/kg) sedation. The animals were examined at autopsy to attribute death to either intracranial tumor growth or normal brain toxicity in the absence of discernible tumor. There was no difference between the two anesthesia groups regarding the survival of unirradiated animals. Radiation therapy produced a significant dose-dependent prolongation in animal survival, which was limited by the development of normal tissue toxicity at the higher doses. When compared to ketamine anesthesia, pentobarbital anesthesia appeared to offer some protection (not statistically significant) against early (but not late) toxicity at selected radiation doses. A reduction in the number of deaths from tissue toxicity suggested an increased antitumor effect, but again this was not statistically significant. Only in one case was there even a marginal significant difference (p = 0.045) between overall therapeutic efficacy in rats sedated with pentobarbital versus ketamine. While there may be a radioprotective effect of pentobarbital in rat brains without intracranial tumor, there is no conclusive evidence for either radioprotection or significant improvement of radiotherapeutic efficacy in this 9L rat brain tumor model. PMID- 8410228 TI - Magnetoencephalographic localization of a language processing cortical area adjacent to a cerebral arteriovenous malformation. Case report. AB - In order to accurately estimate the risk of surgery for dominant perisylvian arteriovenous malformations, the topographical relationship of the lesion to language cortex must be determined. A case is presented in which a magnetoencephalographic (MEG) study was used to map preoperatively and noninvasively an intracortical source of speech-receptive cortex in a 25-year-old right-handed man with a dominant left temporal lobe arteriovenous malformation. The speech-evoked magnetic field was analyzed at 36 positions over the left hemisphere in response to presentations of the consonant-vowel syllables "da" and "ga." A topographical map of the magnetic component evoked at 110 msec after stimulus onset, which was negative going to the vertex in concurrent electrical recordings, was congruent with a superficial cortical neuronal current source. This source was displaced from that usually observed in normal individuals to tonal or click stimuli, being superior to the probable location of auditory cortex, and superior and anterior to the probable location of Wernicke's area as conventionally described. The MEG results were in accord with the determination of position of a language-processing cortical area as assessed by direct electrical stimulation of the cortex during surgery under local anesthesia, and by superselective Amytal (amobarbital) injection during angiography. The MEG recordings and exposed brain stimulation sites were coordinated by cranial measurements, skull x-ray landmarks, and angiographic anatomy. Investigations such as this, which compare MEG findings with those from established clinical procedures, are an essential step in determining the physiological and anatomical utility of magnetoencephalography for noninvasive clinical functional localization. PMID- 8410229 TI - Intracranial vertebral arteriovenous fistula. Case report. AB - True intracranial arteriovenous fistulas are rare. The authors report a case of a direct fistula between the intracranial portion of the vertebral artery and the lateral medullary venous system. The patient initially presented with a subarachnoid hemorrhage. An open surgical approach with clip obliteration of the lesion was used. The anatomy of this lesion and its surgical management are described. PMID- 8410230 TI - True anatomical compartmentalization of the cavernous sinus in a patient with bilateral cavernous dural arteriovenous fistulae. Case report. AB - An unusual case of complete anatomical compartmentalization of the cavernous sinus in a patient with bilateral Type D cavernous dural arteriovenous fistulae is described. This anatomical anomaly isolated the anterior cavernous sinus and orbital venous system, which was primarily responsible for the patient's clinical presentation. The compartmentalization of the cavernous sinus also limited options for definitive endovascular therapy to a transvenous approach via the superior ophthalmic vein. PMID- 8410231 TI - Bilateral temporal bone encephaloceles after cranial irradiation. Case report. AB - Irradiation of the central nervous system may cause significant morbidity, including endocrine dysfunction and intellectual impairment. The authors report a case of bilateral temporal bone encephaloceles in a 21-year-old man who had received prophylactic central nervous system irradiation for acute lymphocytic leukemia in early childhood. Endaural encephaloceles are uncommon, and most occur as a complication of mastoid surgery. The etiology, clinical features, radiological diagnosis, and surgical treatment of temporal bone encephaloceles are discussed. PMID- 8410232 TI - Use of recombinant human erythropoietin to avoid blood transfusion in a Jehovah's Witness requiring hemispherectomy. Case report. AB - The use of perioperative human recombinant erythropoietin is described in a Jehovah's Witness patient. Despite significant anemia, the child's hematocrit was sufficiently increased by the use of erythropoietin so that a two-stage hemispherectomy could be performed without blood transfusion. PMID- 8410233 TI - Primary (granulomatous) angiitis of the central nervous system with multiple aneurysms of spinal arteries. Case report. AB - A 55-year-old woman with primary (granulomatous) angiitis of the central nervous system in association with non-Hodgkin's lymphoma (mucosa-associated lymphoid tissue type) presented with an acute spinal subdural hemorrhage secondary to rupture of one of several fusiform inflammatory aneurysms of the spinal cord radicular arteries. The literature on hemorrhagic complications, aneurysms, and spinal cord involvement in granulomatous angiitis is reviewed. Recognition of granulomatous angiitis is important, as the condition may be responsive to immunosuppressive therapy. PMID- 8410234 TI - The sequential hook insertion technique for universal spine instrumentation application. Technical note. AB - A new technique of universal spine instrumentation insertion for the management of thoracic and lumbar spine instability is presented and the results in 10 patients are described. The technique involves the sequential insertion of Texas Scottish Rite Hospital (TSRH) central-post hooks, followed by hook fixation to the rod; force is then applied with correction of deformity, if needed. This allows for methodical, safe, and rapid instrumentation insertion. The new TSRH central-post hook configuration permits manipulation of the hook/rod relationships to the advantage of the surgeon (and patient) by providing more room for both hook insertion and hook/rod fixation. This technique has reduced operative time, facilitated ease of deformity correction, and provided uniformly acceptable early postsurgical results. PMID- 8410235 TI - A new device for cortical stimulation mapping of surgically unexposed cortex. Technical note. AB - Intraoperative functional mapping is an integral facet of many supratentorial procedures involving cortical resection. A new device is described that can be used in conjunction with a standard cortical stimulator and strip electrodes to stimulate intraoperative regions of unexposed cerebral cortex. This permits the use of a smaller craniotomy, while providing the ability for functional mapping of large areas of surface cortex. PMID- 8410236 TI - Supraorbital craniotomy by fracture of the anterior orbital roof. Technical note. AB - The authors describe a new and rapid method to safely perform a supraorbital craniotomy. This technique can be used when tumor does not invade the orbital roof. Previous descriptions of the supraorbital craniotomy involved exposure of the frontal sinus by removing its anterior wall and using the Gigli saw to separate the orbital roof. This new approach avoids removal of the anterior sinus wall and separates the supraorbital bone flap from the calvaria by fracturing the anterior orbital roof forward. In addition, a method for harvesting a laterally based pericranium and muscle pedicle that contains a section of contralateral temporalis muscle is described. This vascularized pedicle can be used for repair of cerebrospinal fluid leaks or bone defects along the anterior fossa floor and orbit. PMID- 8410237 TI - Neurosurgeon as innovator: William V. Cone (1897-1959). AB - Neurosurgeons are well known for being productive researchers and innovators. Few, however, have possessed the prolific ingenuity of William Cone. In 1934, he and William Penfield were cofounders of the Montreal Neurological Institute where, until 1959, he filled the twin roles of neurosurgeon-in-chief and neuropathologist. Because he did not find writing easy, many of his technical inventions and refinements remained unpublished. His numerous innovations included the extensive use of twist-drill technique for biopsy, drainage for subdural hematoma and cerebral abscess, and ventriculography. In the mid-1940's, he developed power tools driven by nitrogen that led to the modern, universally used air-driven tool systems. He had a special interest in the treatment of spinal dysfunction, for which he invented the Cone-Barton skull-traction tongs along with the Cone spinal operating table. He also devised operative procedures for vertebral fracture-dislocation and craniospinal anomalies. For the maintenance of muscle tone in the paralyzed bladder, he constructed a tidal drainage system. He introduced and popularized ventriculoperitoneal shunting techniques and carried out some of the earliest experimental trails to treat brain infections with sulphonamide and antibiotic drugs. He designed his own set of surgical suction devices, bone rongeurs, and a personal suction "air conditioning" system for each surgeon. He had a keen early interest in intracranial tumors, and also demonstrated on monkeys how subdural mass lesions caused pupillary dilation and mesial temporal lobe damage due to cerebral compression. His work for the military during World War II on effects of altitude on brain pressure remained classified for many years. The first clipping and excision of an intracranial aneurysm is attributed to Cone. Although Penfield was known as "the Chief," Cone was referred to as "the Boss." His fervent dedication to provide total care to his patients was expressed in round-the-clock vigils; he did not separate "nursing" from "surgical" care. Ultimately, Cone's driving passion for perfection led in part to his tragic death. His accomplishments, inventions, and his example as teacher and physician have become part of neurosurgery's collective legacy. PMID- 8410238 TI - Androgen and progesterone receptors in meningiomas. PMID- 8410239 TI - Vagus stimulator for seizures. PMID- 8410240 TI - Head injury and shock. PMID- 8410241 TI - Hypervolemic therapy and vasospasm. PMID- 8410242 TI - Microsurgery and radiosurgery in brain arteriovenous malformations. PMID- 8410243 TI - Microsurgery for 67 intracranial arteriovenous malformations less than 3 cm in diameter. AB - The surgical outcome in a series of small arteriovenous malformations (AVM's) that might have been considered optimal for radiosurgery is reviewed. In a total microsurgical series of 360 patients, 67 (19%) underwent resection of AVM's less than 3 cm in largest diameter, regardless of location. Many of these lesions (45%) were in locations that might be considered surgically inaccessible such as the thalamus, brain stem, medial hemisphere, and paraventricular regions. Complete angiographic obliteration of the AVM by microsurgical technique was accomplished in 63 patients (94%) with a surgical morbidity of 1.5% and no operative mortality. Patients with hemispheric AVM's had a cure rate of 100% and no neurological morbidity. Stereotactically guided craniotomy was used in 14 patients (21%) to locate and resect deep or concealed malformations. The results from five major radiosurgery centers treating similar-sized AVM's are analyzed. The authors' surgical results compare favorably with those from radiosurgery centers which, in their opinion, supports the conclusion that microneurosurgery is superior to radiosurgery, except for a small percentage of lesions that are truly inoperable on the basis of inaccessibility. PMID- 8410244 TI - Stereotactic radiosurgery for cerebral metastatic melanoma. AB - To determine local tumor control rates and survival of patients with melanoma metastases to the brain, the authors reviewed the results of 23 consecutive patients with a total of 32 tumors (19 patients had a solitary tumor and four had multiple tumors) who underwent adjuvant stereotactic radiosurgery. Tumor locations included the cerebral hemisphere (24 cases), brain stem (four cases), basal ganglia (two cases), and cerebellum (two cases). Fifteen patients had associated cranial symptomatology and eight had incidental metastases. All patients had tumors of 3 cm or less in diameter (mean tumor volume 2.5 cu cm), and all received fractionated whole-brain radiation therapy (30 Gy) in addition to radiosurgery (mean tumor margin dose 16 Gy). Nineteen patients were managed with both modalities at the time of diagnosis; four underwent radiosurgery 3 to 12 months after fractionated whole-brain radiotherapy. The mean patient follow-up period was 12 months (range 3 to 38 months). After radiosurgery, eight patients improved, 13 remained stable, and two deteriorated. One patient subsequently required craniotomy because of intratumoral hemorrhage; this patient and three others are living 13 to 38 months after radiosurgery. Nineteen patients died, 18 from progression of their systemic disease and one from another hemorrhage into a new brain metastasis. The local tumor control rate was 97%. Only two patients subsequently developed new intracranial metastases. The median survival period after diagnosis was 9 months (range 3 to 38 months). The authors believe that stereotactic radiosurgery coupled with fractionated whole-brain irradiation is an effective management strategy for cerebral metastases from a melanoma. Multi institutional trials are warranted to confirm that stereotactic radiosurgery results equal or surpass the outcome achieved with craniotomy and tumor resection. PMID- 8410245 TI - Variables affecting the accuracy of stereotactic localization using computerized tomography. AB - Stereotactic localization using computerized tomography (CT) is increasingly employed to guide neurosurgical procedures in crucial areas of the brain such as the brain stem. This technique allows the surgeon to resect a lesion in its entirety while sparing critical areas of the brain. Thus, the parameters used for scanning should be selected for maximum accuracy. While the small pixel size of CT scanners suggests a high degree of precision in localization, there have been few systematic studies of this accuracy. The authors have studied the amount of error in localization created by variables such as CT scan thickness, interscan spacing, size of lesion, and method of computation when using the Brown-Roberts Wells (BRW) stereotactic system. Over 1000 CT scans were made of a phantom composed of spheres of differing diameter and location. The CT slice thickness was varied from 1.5 to 5.0 mm, and interscan spacing was varied from 0.5 to 3.0 mm. The coordinates of the center of the spheres were calculated independently using the laptop computer supplied with the unit and also by a stereotactic computer which automatically calculates the center of the fiducials. The actual BRW coordinates of the sphere center were obtained using the phantom base and were then compared to the computer-calculated coordinates to determine error in localization. Variables with a significant effect on error included the scan thickness, interscan spacing, and sphere size. The mean error decreased 23% as the scan thickness decreased from 5.0 to 1.5 mm and 45% as the interscan spacing decreased from 3.0 to 0.5 mm. Mean error was greatest for the smallest sphere sizes. The two computational methods did not differ in error. This study suggests that, for critical areas of the brain or for small lesions, a scan thickness of 1.5 mm and interscan spacing of 0.5 mm should be employed. PMID- 8410246 TI - Surgery for angiographically occult cerebral aneurysms. AB - In 15% of patients with spontaneous subarachnoid hemorrhage (SAH), the source of bleeding cannot be determined despite repeated cerebral angiography. However, some patients diagnosed as having "SAH of unknown cause" actually harbor undetected aneurysms. The authors report six patients with SAH who, despite multiple negative cerebral angiograms, underwent exploratory surgery due to a high clinical and radiographic suspicion for the presence of an aneurysm. Brain computerized tomography (CT) scans revealed blood located mainly in the basal frontal interhemispheric fissure in four patients, in the sylvian fissure in one patient, and in the interpeduncular cistern in one patient. The patients were evaluated as Hunt and Hess Grades I to III, and had undergone at least two high quality cerebral angiograms that did not reveal an aneurysm. Vasospasm was visualized in two patients. Three patients rebled while in the hospital. Exploratory surgery was performed at an average of 12 days post-SAH. Five aneurysms were discovered at surgery and were successfully clipped. All four patients with interhemispheric blood were found to have an anterior communicating artery (ACoA) aneurysm. The patient with blood in the sylvian fissure was found to have a middle cerebral artery aneurysm. These aneurysms were partially thrombosed. No aneurysm was detected in the patient with interpeduncular SAH, despite extensive basilar artery exploration. Five patients had an excellent outcome and one patient developed diabetes insipidus. These results show that exploratory aneurysm surgery is warranted, despite repeated negative cerebral angiograms, if the patient manifests the classical signs of SAH with CT scans localizing blood to a specific cerebral blood vessel (particularly the ACoA) and if a second SAH is documented at the same site. PMID- 8410247 TI - Partial sensory trigeminal rhizotomy at the pons for trigeminal neuralgia. AB - Microvascular decompression is preferred among open procedures for the treatment of trigeminal neuralgia. However, in some cases the decompression cannot be performed, either because no significant vascular compression of the trigeminal nerve is found at surgery or because a patient's vascular anatomy makes it unsafe. Partial sensory rhizotomy is a commonly used alternative in these instances. The outcome after partial sensory rhizotomy was reviewed retrospectively in 83 patients with an average follow-up period of 72 months. Sixty-four (77%) of these patients had no evidence of vascular contact at operation. The remaining 19 patients (23%) had vascular structures in proximity to the trigeminal nerve but still underwent partial sensory rhizotomy in place of or in addition to microvascular decompression either because the offending vessel could not be moved adequately (11 cases) or because the vascular contact was considered insignificant (eight cases). Outcome was classified as: excellent if there was no trigeminal neuralgia postoperatively; good if pain persisted or recurred but was less severe than preoperatively; and poor if persistent or recurrent pain was equal to or greater than the preoperative pain in severity and was refractory to medication, or was severe enough to require additional surgery. The outcome was excellent in 40 patients (48%), good in 18 (22%), and poor in 25 (30%); follow-up durations were similar for the three outcome categories. The failure rate was 17% for the 1st year and averaged 2.6% each year thereafter. Two variables were predictive of a poor outcome: prior surgery and lack of preoperative involvement of the third trigeminal division. Major complications occurred in 4% of cases and minor complications in 11%. The authors conclude that partial sensory rhizotomy is a safe and effective alternative to microvascular decompression when neurovascular compression is not identified at operation or when microvascular decompression cannot be performed for technical reasons. PMID- 8410248 TI - Corpus callosotomy: a quantitative study of the extent of resection, seizure control, and neuropsychological outcome. AB - Corpus callosotomy is valuable for controlling medically intractable generalized seizures in appropriate patients, but postoperative development of language disorders, neuropsychological impairment, and motor dysfunction have all been noted. The extent of callosum resection has been implicated as a possible determinant of outcome, but this hypothesis has not been formally tested. Analysis of the records of all patients who underwent corpus callosotomy at the University of California, San Francisco, from 1986 to 1991 showed that, of 15 patients who underwent anterior or complete callosotomy, seven were entirely or nearly seizure-free, four had at least a 50% reduction in seizure frequency, and four had no change. To determine callosal size and extent of callosotomy, preoperative and postoperative magnetic resonance images were measured with computer-based planimetry. Seizure outcome was not significantly associated with preoperative callosal size or extent of callosotomy. Intelligence quotient scores did not change significantly after callosotomy. No severe neuropsychological deficits developed after anterior or complete callosotomy, even in patients with mixed cerebral dominance or bilateral language representation. These results indicate that division of the anterior one-half to two-thirds of the corpus callosum is nearly as effective as more extensive anterior sectioning or complete callosotomy in reducing drop-attack and generalized tonic-clonic seizures in appropriate patients, and that the extent of callosotomy is not an important factor on outcome when at least 50% to 65% of the callosum is divided. Mixed cerebral dominance and other unusual patterns of language and memory organization do not appear to increase the postoperative risk for neuropsychological deficits, regardless of the extent of anterior section. PMID- 8410249 TI - Catecholamine response to a gradual increase of intracranial pressure. AB - To determine the catecholamine response to progressive intracranial hypertension, intracranial pressure (ICP) was raised gradually by continuous expansion of an epidural balloon in seven dogs. Hemodynamic parameters, ICP, and cerebral perfusion pressure (CPP) were monitored continuously and serum catecholamine levels began to rise when CPP was in the low-positive range (20 to 30 mm Hg), reaching a peak just after brain death (CPP < or = o mm Hg). There was no correlation between ICP and the catecholamine peak. Compared to control values, the mean increase was 286-fold for epinephrine and 78-fold for norepinephrine. Temporally, the catecholamine peak corresponded well with the observed hemodynamic changes. These results suggest that ischemia in certain parts of the brain stem is responsible for the hemodynamic changes observed in intracranial hypertension (such as the Cushing response), and they show that catecholamines play an important role in these hemodynamic changes. PMID- 8410250 TI - Therapeutic effects of local delivery of dexamethasone on experimental brain tumors and peritumoral brain edema. AB - To determine if dexamethasone administered by osmotic pump directly to brain tumors would control peritumoral edema and at the same time suppress tumor growth and prolong survival, the authors studied experimental brain tumors produced in 102 rabbits by implanting VX2 carcinoma cells. Of these, 58 animals were separated into three treatment groups: Group 1 included 15 untreated rabbits; Group 2 included 18 rabbits treated with systemic dexamethasone (4 mg/kg/day); and Group 3 included 25 rabbits treated with local dexamethasone (0.24 mg/day) delivered by osmotic pump. Systemic or local dexamethasone was administered from Day 3 or Day 7 after tumor implantation, and animals were sacrificed on Day 13. A survival study was performed with 44 rabbits separated into the same treatment groups, beginning drug delivery on Day 7. Brain water content in the white matter of sacrificed animals was measured by the specific gravity method. The length and width of the brain tumors in all animals were measured and the tumor volume estimated. Findings showed that systemic and local dexamethasone administered from Day 3 or Day 7 was associated with a significant (5% level) inhibition of tumor volume as well as a mean reduction of brain edema in most tested sites. Systemic and local dexamethasone therapy also resulted in a significant (5% level) increase in survival time relative to the untreated group. These short term results suggest that locally delivered dexamethasone may constitute a clinically important therapeutic modality. PMID- 8410251 TI - Altered response to histamine in brain tumor vessels: the selective increase of regional cerebral blood flow in transplanted rat brain tumor. AB - The authors studied the effect of intracarotid administration of histamine on the regional cerebral blood flow (rCBF) in transplanted rat C6 glioma by the hydrogen clearance method. Histamine infusion at doses of 1 and 10 micrograms/kg/min produced an increase of rCBF in the tumor (24.6% +/- 16.4%, p < 0.002, and 37.6% +/- 18.2%, p < 0.0001, respectively) and also in brain surrounding the tumor (26.8% +/- 16.2%, p < 0.002, and 34.9% +/- 9.2%, p < 0.0001, respectively) without any significant changes in the ipsilateral hemisphere. Intravenous administration of pyrilamine (H1 antagonist) and cimetidine (H2 antagonist) reduced blood flow responses to histamine; cimetidine was a more effective blocking agent than pyrilamine. Intracarotid infusion of histamine (1 and 10 micrograms/kg/min) with intravenous injection of Evans blue dye disclosed the selective extravasation of dye in the tumor and the brain surrounding the tumor. These results indicated that brain tumor vessels could respond to histamine differently than normal brain capillaries. The mechanism of selective response to histamine could be explained either by increased permeability or by altered characteristics of histamine receptors in the tumor vessels. PMID- 8410252 TI - Thymidine kinase-mediated killing of rat brain tumors. AB - Gene therapy has many potential applications in central nervous system (CNS) disorders, including the selective killing of tumor cells in the brain. A rat brain tumor model was used to test the herpes simplex virus (HSV)-thymidine kinase (TK) gene for its ability to selectively kill C6 and 9L tumor cells in the brain following systemic administration of the nucleoside analog ganciclovir. The HSV-TK gene was introduced in vitro into tumor cells (C6-TK and 9L-TK), then these modified tumor cells were evaluated for their sensitivity to cell killing by ganciclovir. In a dose-response assay, both C6-TK and 9L-TK cells were 100 times more sensitive to killing by ganciclovir (median lethal dose: C6-TK, 0.1 microgram ganciclovir/ml; C6, 5.0 micrograms ganciclovir/ml) than unmodified wild type tumor cells or cultured fibroblasts. In vivo studies confirmed the ability of intraperitoneal ganciclovir administration to kill established brain tumors in rats as quantified by both stereological assessment of brain tumor volumes and studies of animal survival over 90 days. Rats with brain tumors established by intracerebral injection of wild-type or HSV-TK modified tumor cells or by a combination of wild-type and HSV-TK-modified cells were studied with and without ganciclovir treatments. Stereological methods determined that ganciclovir treatment eliminated tumors composed of HSV-TK-modified cells while control tumors grew as expected (p < 0.001). In survival studies, all 10 rats with 9L-TK tumors treated with ganciclovir survived 90 days while all untreated rats died within 25 days. Curiously, tumors composed of combinations of 9L and 9L-TK cells could be eliminated by ganciclovir treatments even when only one-half of the tumor cells carried the HSV-TK gene. While not completely understood, this additional tumor cell killing appears to be both tumor selective and local in nature. It is concluded that HSV-TK gene therapy with ganciclovir treatment does selectively kill tumor cells in the brain and has many potential applications in CNS disorders, including the treatment of cancer. PMID- 8410253 TI - Epidural perfusion cooling protection against protracted spinal cord ischemia in rabbits. AB - The protective effect of a modified epidural cooling technique was assessed in a rabbit spinal cord ischemia model. The epidural space around the lumbar segments with induced ischemia was continually perfused with cold (5 degrees C) isotonic saline via two communicating spinal canal openings. This procedure allowed the spinal cord to be kept deeply hypothermic (< 15 degrees C within central gray matter) during the ischemic period. The animals were subjected to either normothermic ischemia (Group A) or hypothermic ischemia (Group B). Each group contained three subgroups of animals undergoing 20, 40, or 60 minutes of aortic ligation. Their neurological outcomes were evaluated up to 48 hours postischemia, and the intergroup differences were compared. Two days postischemia, all of the animals were sacrificed by transcardial perfusion-fixation and their lumbar segments were processed for histopathological examination. In addition, in animals with 60-minute ischemia, spinal somatosensory evoked potentials were recorded during surgical intervention and again after 48 hours. In the normothermic animals, a high incidence of paraplegia was detected: in 40% after 20 minutes of ischemia, in 75% after 40 minutes, and in 100% after 60 minutes. In contrast, all of the hypothermic animals exhibited full neurological recovery even after 60 minutes of ischemia. Both electrophysiological and histological observations clearly correlated with the neurological findings. The results suggest that deep spinal cord hypothermia produced by epidural perfusion cooling provides effective protection against protracted spinal cord ischemia in rabbits. PMID- 8410254 TI - Limiting ischemic spinal cord injury using a free radical scavenger 21 aminosteroid and/or cerebrospinal fluid drainage. AB - Traumatic spinal cord injury occurs in two phases: biomechanical injury, followed by ischemia and reperfusion injury. Biomechanical injury to the spinal cord, preceded or followed by various pharmaceutical manipulations or interventions, has been studied, but the ischemia/reperfusion aspect of spinal cord injury isolated from the biomechanical injury has not been previously evaluated. In the current study, ischemia to the lumbar spinal cord was induced in albino rabbits via infrarenal aortic occlusion, and two interventions were analyzed: the use of U74006F (Tirilazad mesylate), a 21-aminosteroid, and cerebrospinal fluid (CSF) drainage. These treatment modalities were tested alone or in combination. In Phase 1 of this study, the rabbits received 1.0 mg/kg of Tirilazad or an equal volume of vehicle (controls) prior to the actual occlusion, three doses of Tirilazad (1 mg/kg each) during the occlusion, then several doses after the occlusion. Of the Tirilazad-treated animals, 30% became paraplegic while 70% of the control animals became paraplegic. Phase 2 involved the same doses of Tirilazad as in Phase 1 and, in addition, CSF pressure monitoring and drainage were performed. The paraplegia rate was 79% in the control animals, 36% in the group receiving Tirilazad alone, 25% in the group with CSF drainage alone, and 20% in the Tirilazad plus CSF drainage group. This rate also correlated with changes noted in CSF pressure; both Tirilazad administration alone and CSF drainage alone induced a decrease in CSF pressure and the two combined produced a further decrease. There was marked improvement in the perfusion pressure when using Tirilazad alone, CSF drainage alone, and Tirilazad therapy in combination with CSF drainage, with the last group producing the largest increase. This change in CSF pressure and perfusion pressure correlated with improved functional neurological outcome. Pathological examination revealed that Tirilazad therapy reduced the extensive and diffuse neuronal, glial, and endothelial damage to (in its most severe form) a more patchy focal region of damage in the gray matter. Cerebrospinal fluid drainage resulted in pyknosis of some motor neurons, and some eosinophilia. The combination of CSF drainage and Tirilazad administration resulted in the least abnormality, with either normal or near-normal spinal cords. It is concluded that Tirilazad administration decreased CSF pressure during spinal cord ischemia and reperfusion and, like CSF drainage, increased and improved the perfusion pressure to the spinal cord, decreased spinal cord damage, and improved functional outcome.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410255 TI - Dissociation of cerebral blood flow, glucose metabolism, and electrical activity in pediatric brain death. Case report. AB - A 2-month-old infant demonstrated clinical brain death 48 hours after suffering a closed head injury accompanied by cardiac arrest. Two nuclear cerebral blood flow (CBF) studies demonstrated normal perfusion. On the 11th day following injury, cerebral electrical activity ceased and a normal glucose metabolic gradient between gray and white matter was documented on positron emission tomography. Autopsy revealed widespread necrosis with mononuclear cell infiltrates throughout all cerebral cortical layers. Nine children have previously been described with clinical brain death, electrocerebral silence, and evidence of CBF by radionuclide scan. The dissociation between cerebral electrical activity and blood flow may be explained by an increase in cranial volume allowed by the expansile neonatal skull, preventing both intracranial hypertension and a reduction in perfusion pressure. The persistence of glucose metabolism may be associated with the presence of inflammatory microglial cells in the ischemic cortex. The authors conclude that persistence of CBF and glucose metabolism in brain-dead children may not indicate neuronal survival. If repeated neurological examinations with or without electroencephalography support the diagnosis of brain death, the presence of CBF and glucose metabolism should not alter this conclusion. PMID- 8410256 TI - Superficial siderosis of the central nervous system: magnetic resonance imaging and pathological correlation. Case report. AB - The authors report a 32-year-old woman who had undergone repair of an occipital encephalocele in infancy and who experienced a 20-year history of progressive hearing loss and intermittent vertigo. After parturition, she developed a rapidly progressive quadriparesis and brain-stem dysfunction associated with persistent intraventricular and subarachnoid hemorrhage. Serial magnetic resonance (MR) images showed progressive deposition of hemosiderin along the surface of the brain, brain stem, and spinal cord, and enhanced thickened membranes at the site of the original encephalocele repair. Posterior fossa exploration disclosed hemorrhagic membranes, which were resected; despite removal of this tissue, the patient deteriorated and died. Postmortem examination confirmed iron-containing pigment along the meninges, cerebral hemispheres, brain stem, spinal cord, and cranial nerves accompanied by atrophy of the superficial cerebellar cortex. It is concluded that superficial siderosis may accompany encephalocele repair. This is believed to be the first report in the literature of superficial siderosis of the central nervous system to correlate in vivo MR images with autopsy results. PMID- 8410257 TI - Increased mitotic activity as a negative prognostic indicator in pleomorphic xanthoastrocytoma. Case report. AB - Pleomorphic xanthoastrocytoma is a recently characterized neoplasm with a favorable prognosis despite aggressive histological features. The authors report a case of pleomorphic xanthoastrocytoma that recurred 4 years after complete gross resection. The original tumor exhibited histological features characteristic of this neoplasm, but up to 4 mitoses/10 high-power fields were present focally. The recurrent tumor contained small foci of classical pleomorphic xanthoastrocytoma, but consisted predominantly of glioblastoma multiforme. Transitional zones contained nests of glial fibrillary acidic protein (GFAP)-immunopositive cells surrounded by delicate collagenous and reticulin-rich septa. Electron microscopy of the transitional zone showed continuous basal lamina investing cells containing bundles of intermediate filaments. These were GFAP-positive by immunogold electron microscopy, confirming the astrocytic nature of pleomorphic xanthoastrocytoma. This example illustrates the capacity of this tumor to evolve into glioblastoma. The indolent clinical behavior of most pleomorphic xanthoastrocytomas is evident from a literature review, which confirms the prolonged survival of many patients after onset of symptoms. Completeness of excision, subjectively assessed at surgery, did not influence the risk of recurrence or survival up to 10 years after initial resection. Postoperative radiotherapy did not improve survival, but may reduce the probability of recurrence; more studies are needed to corroborate this finding. The data compiled herein support the designation of pleomorphic xanthoastrocytoma as a distinct astrocytic neoplasm with a favorable prognosis. An increased mitotic rate has not previously been correlated with a worse outcome, and should not be used to exclude this diagnosis. However, anaplastic transformation of pleomorphic xanthoastrocytoma confers a much worse prognosis, and this case suggests that increased mitotic activity may be a negative prognostic indicator since it may herald subsequent anaplastic transformation. PMID- 8410258 TI - Isolated extranodal intracranial sinus histiocytosis in a 5-year-old boy. Case report. AB - Sinus histiocytosis with massive lymphadenopathy was first described in 1969 by Rosai and Dorfman. The typical clinical characteristics of this disease include painless cervical lymphadenopathy, fever, and weight loss. The condition can present with an extranodal mass in about 25% of patients, and isolated masses without lymph node involvement occur rarely. The authors describe a 5-year-old boy with cavernous sinus syndrome due to an isolated extranodal form of sinus histiocytosis with massive lymphadenopathy in the temporal fossa. Several cases of this disease involving the central nervous system are reviewed. The histopathological and magnetic resonance imaging characteristics are discussed. PMID- 8410259 TI - Localized hypertrophic mononeuropathy involving spinal roots and associated with sacral meningocele. Case report. AB - Hypertrophic nerve lesions displaying onion-bulb cellular formations are quite rare in the absence of a generalized hypertrophic neuropathy. The isolated peripheral nerve lesion has been termed "localized hypertrophic mononeuropathy" (LHN), and fewer than 30 cases of this condition have been reported. Very little is known regarding the etiology and the natural course of this rare disorder. A unique case of LHN afflicting spinal roots in association with a sacral meningocele is reported with a brief review of the relevant literature. The unique features of this case not only reveal a variable clinical presentation of the disease but also support the theory that LHN may be a localized reaction to nerve trauma or entrapment. PMID- 8410260 TI - Carotid artery revascularization following Crutchfield clamp placement. Report of two cases. AB - Several types of adjustable clamp have been widely utilized to gradually occlude the carotid artery for the treatment of various intracranial vascular lesions. A fairly large number of patients, many of whom have not been adequately followed, have these clamps still in place. The authors report two patients, initially treated with a Crutchfield clamp for an intracranial aneurysm, in whom carotid artery system revascularization occurred through the clamp many years later, leading to continued filling of the aneurysm. Recommendations are given on monitoring patients with Crutchfield clamps in order to minimize long-term complications. PMID- 8410261 TI - Split-calvaria osteoplastic rotational flap for anterior fossa floor repair after tumor excision. Technical note. AB - A split-calvaria osteoplastic rotational flap to repair the anterior fossa floor after tumor excision was devised and tested clinically. At surgery, the flap is outlined between the glabella and the vertex. After the pericranium between the glabella and the flap's anterior pole is elevated to form its pedicle, a full thickness craniotomy is performed to expose the diploic aspect of the bone graft donor site (when the graft is relatively wide, bifrontal craniotomies may be advantageous). The diploic space is split in situ, taking care to protect the pedicle and its attachments to the osseous segment. Linear osteotomies in the outer table are created to mobilize the flap. With the flap rotated frontally, the craniotomy is completed. After tumor extirpation, the margins of the osseous segment of the flap are shaped to conform to the defect of the anterior fossa floor. Transverse osteotomies are performed so that the graft's convex curve conforms to that of the anterior fossa floor. The flap is then rotated into position. Follow-up evaluation in two patients at 22 and 30 months demonstrated bone integrity of the anterior fossa floor with graft preservation. Transient postoperative cerebrospinal fluid (CSF) rhinorrhea, which occurred in Case 1, was avoided in Case 2 by placing the osseous segment of the graft coplanar with the bone floor of the fossa. Neither patient had late meningitis or CSF rhinorrhea. The split-calvaria osteoplastic rotational flap may represent an advance toward the ideal reconstruction of the anterior cranial fossa floor. PMID- 8410262 TI - New transducers for intraoperative spinal sonography. Technical note. AB - Although ultrasonography during spinal cord surgery has been widely accepted, it still has limitations in clinical application because of the size of the transducers needed and the immersion method. The authors report on newly designed microprobe transducers used to evaluate intramedullary spinal cord lesions and to monitor surgical procedures. This microprobe system permitted the microscopic use of intraoperative ultrasonography without requiring saline immersion. PMID- 8410263 TI - Precise terminology in scientific communications. PMID- 8410264 TI - Microsurgery and radiosurgery in small AVM's. PMID- 8410265 TI - Intra-arterial papaverine for vasospasm. PMID- 8410266 TI - Complications of hypervolemic therapy. PMID- 8410267 TI - Complications of hypervolemic therapy. PMID- 8410268 TI - Intracerebral polyposis or osteoma? PMID- 8410269 TI - Intracerebral polyposis or osteoma? PMID- 8410270 TI - Drilling the posterior meatal lip. PMID- 8410271 TI - Indium-111-labeled polyclonal human immunoglobulin: identifying focal infection in patients positive for human immunodeficiency virus. AB - Pooled human immunoglobulin labeled with indium-111 (111In-HIgG) was used to identify the presence and extent of infection in patients positive for human immunodeficiency virus (HIV), presenting with either symptoms and/or signs of acute chest infection or with pyrexia without localizing signs or symptoms. Fifty five studies were performed in 51 patients with suspected chest infection or pyrexia without localizing signs. Of these, 111In-HIgG identified intrapulmonary accumulation in 17 patients with Pneumocystis carinii pneumonia, eight with bacterial pneumonia, five with cytomegalovirus pneumonia, three with pulmonary Mycobacterium avium intracellulare infection and one with a fungal pneumonia. There was no intrapulmonary accumulation of 111In-HIgG in five patients with bronchopulmonary Kaposi's sarcoma and in three patients with intrathoracic lymphoma. Quantification of lung/heart activity was significantly increased (p < 0.05) in patients with active chest infection compared with those with intrapulmonary tumor or no active lung pathology. Indium-111-HIgG scintigraphy also localized at 14 sites of extrapulmonary infection, including six patients with colitis. There were no false-negative studies but false-positive uptake was seen in four studies. These results confirm that 111In-HIgG correctly identifies the presence and extent of infection in patients positive for HIV antibody. PMID- 8410272 TI - Different features of pulmonary metastases in differentiated thyroid cancer: natural history and multivariate statistical analysis of prognostic variables. AB - We studied 134 patients with differentiated thyroid cancer and pulmonary metastases. All were treated with total or near total thyroidectomy, radioiodine and L-thyroxine. The prognostic value of the following variables in three groups of patients were evaluated by univariate and multivariate analysis: age at diagnosis, sex, histologic type, tumor extension, cervical lymph node metastases, mediastinic metastases, presence of metastases in distant sites other than lungs (multiple distant metastases) and morphological (chest x-rays) and functional (131I uptake) features of lung metastases. Univariate analysis identified patient age (p < 0.0001), morphological and functional features of lung metastases (p < 0.0001), presence of multiple distant metastases (p < 0.0001) and histologic type (p = 0.04) as significant prognostic factors. Multivariate analysis showed only morphological (p = 0.0014) and functional (p < 0.0001) features of lung metastases and the presence of multiple distant metastases (p = 0.01) as significant and independent variables. The data show that early (pre radiological) scintigraphic diagnosis and 131I therapy of lung metastases appear to be the most important elements in obtaining both a significant improvement in survival rate and a prolonged disease-free time interval in these patients. PMID- 8410273 TI - Optimized dose planning of radioiodine therapy of benign thyroidal diseases. AB - Thyroid uptake measurements were performed on 246 patients, who underwent radioiodine therapy for benign disease up to 192 hr after oral application of either the test activity (7 MBq) or the therapeutic activity (150-1100 MBq). Using the complete set of uptake values, the cumulated activity in the thyroid was calculated and the dose-to-activity ratio (D/A) deduced. An empiric factor was derived, which allows prediction of the D/A with high precision, using only the late uptake measurement at 96 hr or 192 hr. The correlation between the value calculated from the complete set of uptake values and that of only one at 96 hr or 192 hr was R = 0.99 and 0.97, respectively. The activity required for intended dose can thus reliably be determined by a single, late uptake measurement. In a second analysis, the correlation between the D/A of the test and the therapeutic activity was established. There were two essential findings: For those patients who were without or under stable thyroid-specific medication there was a strong correlation between the two D/A values. The therapeutic value was on the average approximately 15% lower than the test ratio, which is assumed to be due to an enhanced iodine turnover under therapeutic conditions. In patients whose medication changed close to the test study or therapy, the measured test and therapeutic D/A were strongly noncorrelated. PMID- 8410274 TI - Optimization of radioiodine therapy of thyrotoxicosis: what have we learned after 50 years? PMID- 8410275 TI - Physiopathological significance of thallium-201 per rectum scintigraphy in liver cirrhosis. AB - To define the physiopathological significance of 201Tl per rectum scintigraphy, we compared results obtained using this method with direct measurement of inferior mesenteric shunting, portal pressure, liver cellular function as evaluated by the Aminopyrine Breath Test and the size of esophagogastric varices and spleen, constituting, respectively, indirect representation of azygos and splenic shunts. Results indicated that a high correlation exists between the measures of portal systemic shunt estimated by the per rectal method and those obtained by direct administration of the tracer in the inferior mesenteric artery. No correlation was observed between 201Tl per rectal results and portal pressure or with azygos and splenic shunting. Fair correlation was observed with the Aminopyrine Breath Test. This could be explained by the fact that both methods are altered in advanced liver disease. These results suggest that the 201Tl per rectal scintigraphy explores the portal systemic shunt, which depends almost exclusively on the inferior mesenteric territory. While the limited territory explored by the method constitutes, undoubtedly, a limiting factor in detection and quantitation of total portal-systemic shunt, the specific information provided by the test could be useful in defining clinical and biological profiles of cirrhotic patients with inferior mesenteric shunting. PMID- 8410276 TI - Technetium-99m-nanocolloid scintigraphy in orthopedic infections: a comparison with indium-111-labeled leukocytes. AB - Twenty-three patients with clinically suspected acute or chronic osteomyelitis and 21 patients with suspected joint prosthetic infection underwent scintigraphy using both 99mTc-nanocolloid and 111In-labeled leukocytes. The scintigrams of the two tracers were blindly interpreted by three independent observers. Their evaluations showed high correspondence. Patients were classified as having no infection, probable infection or proven infection according to specific criteria which included results of bacteriological cultures and histopathological examinations. For proven and probable infection taken together, the sensitivity with 99mTc-nanocolloid was 94%, the specificity 84% and the accuracy 87%, compared with 75%, 90% and 85% with 111In-labeled leukocytes. We conclude that 99mTc-nanocolloid scintigraphy is at least equivalent with 111In-leucocyte scintigraphy, and its additional advantages are shorter examination time, less complexity and better radiation dosimetry. PMID- 8410277 TI - Clinical immunoscintigraphy of ovarian carcinoma using iodine-131-labeled 145-9 monoclonal antibody. AB - The monoclonal antibody (Mab) designated 145-9 recognizes CA125 antigen but binds to a different epitope than that recognized by OC125 antibody. This is a clinical study assessing the safety, kinetics and imaging sensitivity of Mab 145-9. Two milligrams of Mab were labeled with 111 MBq (3.0 mCi) of 131I and infused intravenously in 18 patients with ovarian carcinoma. Immunoscintigraphies were done at three, five, and seven days. There were no adverse reactions to the injection of this Mab. All immunoscintigraphies were considered positive. Immunoscintigraphy detected tumor lesions were confirmed in operative fields, in two patients with normal serum levels of CA125 and in four patients whose sonography and/or x-ray computed tomography showed negative findings. In five patients, immunoscintigraphy was repeated without any adverse reaction and revealed the progress of the carcinoma. Pharmacokinetic studies showed the steady state volume of distribution (Vdss) to be 2772 +/- 466 ml (mean +/- s.d.), and clearance 51.3 +/- 12.7 ml/hr. In summary, immunoscintigraphies using 131I labeled Mab 145-9 were done safely in patients with ovarian carcinoma. Preliminary results reveal a high sensitivity compared to radiological methods and tests currently in use. PMID- 8410278 TI - Lesion-by-lesion comparison of computerized tomography and indium-111-labeled monoclonal antibody C110 radioimmunoscintigraphy in colorectal carcinoma: a multicenter trial. AB - C110 is an anti-carcinoembryonic antigen murine IgG1 monoclonal antibody. Indium 111-labeled C110 radioimmunoscintigraphy (RIS) in colorectal cancer was studied in 51 presurgical patients at four institutions. Planar and SPECT images were obtained at least twice between 48 and 96 hr after injection of 5 mCi/5 mg of 111In-C110. Fifty-one patients had 87 biopsy-proven lesions at surgery (57 hepatic, 30 extra-hepatic). Thirty-three patients (64.7%) had positive radionuclide scans, while 32 (62.8%) had positive CT scans (p = NS). While CT was better at overall lesion detection (62.1% versus 56%, p < 0.05), radionuclide scans were better than CT for extra-hepatic disease (60% versus 46.7%, p < 0.01). Hepatic metastases (52.6%) were visualized by Mab scans to selectively concentrate radioactivity. Uptake in draining lymph nodes was a significant limitation, making evaluation of these sites difficult. Indium-111-C110 shows selective uptake in metastatic colorectal cancer, including more than half of all hepatic lesions evaluated. PMID- 8410279 TI - Bispecific monoclonal antibody-mediated targeting of an indium-111-labeled DTPA dimer to primary colorectal tumors: pharmacokinetics, biodistribution, scintigraphy and immune response. AB - Eleven patients with primary colorectal carcinoma tumors (4 +/- 2 cm) were given intravenous injections of 1-10 mg of an anti-CEA, anti-In-DTPA bispecific Fab' Fab monoclonal antibody, and 2-8 days later, were injected with 1.2-4.2 nmol of an 111In-labeled DTPA dimer (6 mCi). The bispecific antibody exhibited good stability and F(ab)'2-like pharmacokinetics. After injection, the 111In-DTPA dimer distributed in a large volume (88 ml/kg-180 ml/kg) and cleared through the kidneys (mean residence time in the whole body: 9 hr-16 hr). Uptake of 111In by the tumor using this two-step technique (1.8%-17.5% injected dose ID/kg, measured from surgical samples 48 hr after hapten injection) was not found significantly lower than that achieved with our reference 111In-labeled anti-CEA F(ab)'2 1 to 4 days after injection in six patients with similar clinical status (5.5%-30.2% ID/kg). In addition, tumor-to-blood and tumor-to-liver uptake ratios were significantly improved (blood 7.8 versus 4.2, liver 2.8 versus 0.8). As a result, low background images allowed detection of 12 of 13 lesions, 4 hr and 24 hr after hapten injection. However, 7 of 11 patients developed HAMA. PMID- 8410280 TI - PET and the autoradiographic method with continuous inhalation of oxygen-15-gas: theoretical analysis and comparison with conventional steady-state methods. AB - The steady-state method using 15O gas inhalation and positron emission tomography (PET) is a simple and practical way of imaging cerebral blood flow (CBF) and oxygen metabolism. Several disadvantages do exist, however, including prolonged examination time, requirement of steady-state and a large tissue heterogeneity effect. To avoid the drawbacks of the steady-state method but to preserve its simplicity, we applied the PET/autoradiographic method to the build-up phase during the continuous inhalation of 15O-gas with intermittent arterial sampling. A simulation study was performed to determine the optimal scanning period, evaluate the delay and dispersion effect of the input function and estimate the tissue heterogeneity effect. To assess the clinical feasibility of the proposed technique for the study of oxygen metabolism, sequential measurements with this method and the conventional steady-state method were performed in eight patients. The simulation study showed that a 5-min scan started 3 min after the commencement of 15O-gas inhalation was optimal. With this method, the delay and dispersion effect on CBF was the same as that of the conventional steady-state method, but the tissue heterogeneity effect was reduced. In eight patients, CBF values calculated by this method showed time dependency and were slightly higher than those obtained by the steady-state method. The oxygen extraction fraction showed no significant time dependency and was well correlated with that obtained by the steady-state method. We conclude that the proposed method is a simple and acceptable alternative to the conventional steady-state method. PMID- 8410281 TI - Quantitative analysis of PET and MRI data in normal aging and Alzheimer's disease: atrophy weighted total brain metabolism and absolute whole brain metabolism as reliable discriminators. AB - Average whole brain metabolic rates, when corrected for brain atrophy, are similar between patients with Alzheimer's disease (AD) and age-matched controls. To elucidate the relationship between reduced cognitive function and cerebral metabolism in patients with AD, we hypothesized that the absolute amount of glucose used by the entire brain may prove to be a more reliable indicator of the disease than metabolic rates calculated for a unit of brain weight. Twenty patients with the probable diagnosis of AD and 17 similarly aged controls underwent 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) studies as well as magnetic resonance imaging (MRI) within a few days of each other. Average metabolic rates, when corrected for atrophy, were 3.91 +/- 1.02 and 4.43 +/- 0.87 (mg of glucose per 100 cc brain tissue per min +/- s.d.) respectively for AD patients and controls. Two other indices were determined, atrophy weighted total brain metabolism (calculated by multiplying the brain volume, determined by MR analysis, by the average metabolic rate) and absolute whole brain metabolism (calculated by multiplying the brain volume by the average metabolic rate corrected for atrophy). The former showed a very significant difference between the two groups (29.96 +/- 7.90 for AD patients compared to 39.1 +/- 7.0 for controls, p < 0.001). Atrophy weighted total brain metabolism also correlated very well with mini mental status exam (MMSE) scores (r = 0.59, p < 0.01). Absolute whole brain metabolism was significantly different between AD and control groups and correlated well with MMSE. These data demonstrate that although the metabolic rate per unit weight of the brain is unchanged in AD compared to controls, atrophy weighted total brain metabolism and absolute whole brain metabolism are significantly affected. Both indices may prove to be a sensitive correlate for cognitive dysfunction in AD. PMID- 8410282 TI - Reverse redistribution in resting thallium-201 myocardial scintigraphy in patients with coronary artery disease: relation to coronary anatomy and ventricular function. AB - We studied 25 male patients, with coronary artery disease, mean age 56 +/- 8 yr. All underwent 201Tl rest-redistribution and resting 99mTc methoxyisobutyl isonitrile (MIBI) cardiac imaging. Regional 201Tl and MIBI uptake were quantitatively analyzed. Regional left ventricular wall motion (WM) was visually assessed on MIBI gated images using a three-point scale (0 = normal, 1 = hypokinetic, 2 = a/dyskinetic). Two patterns of reverse redistribution (RR) were identified: RR-A when 201Tl uptake was normal on rest images and abnormal on redistribution images, and RR-B when 201Tl uptake was abnormal on rest images and a significant decrease in uptake was observed on redistribution images. Of the total 375 myocardial segments analyzed, 229 were classified as normal (Nl), 40 as reversible defect (RD), 74 as irreversible defect (ID); 26 showed RR-A while 6 myocardial segments had RR-B. Myocardial segments with RR-A differed from NI in the degree of coronary artery stenosis (81% +/- 33% versus 57% +/- 39%, respectively, p < 0.05), in WM score (1.1 +/- 0.7 versus 0.5 +/- 0.6, respectively, p < 0.01), and in MIBI uptake (81% +/- 10% versus 92% +/- 9%, respectively, p < 0.0001). Moreover, the percent of myocardial segments supplied by a totally occluded coronary artery was significantly higher (p < 0.05) in myocardial segments with RR-A (46%) than in NI (22%). Segments with RR-B did not show any significant difference either from RD and ID. These results suggest that myocardial segments with RR-A on resting 201Tl images have impaired function and are supplied by severely stenosed coronary arteries and should not be considered normal. PMID- 8410283 TI - Easy come, easy go: time to pause and put thallium reverse redistribution in perspective. PMID- 8410284 TI - Right ventricular thallium-201 kinetics in pulmonary hypertension: relation to right ventricular size and function. AB - Right ventricular ischemia occurs in experimental models of pulmonary hypertension. We analyzed right ventricular size and function and 201Tl uptake to determine if there was a relationship between 201Tl uptake and systolic function in 19 patients with pulmonary artery hypertension who were being evaluated for heart-lung transplantation. All patients had dipyridamole stress 201Tl scintigraphy, radionuclide angiography and echocardiography. In nine patients (Group 1), right ventricular ejection fraction was < 30% (mean 22% +/- 8%). In 10 patients (Group 2) it was > 30% (mean 45% +/- 11%). In Group 1, right ventricular 201Tl uptake in the lateral wall after dipyridamole was increased compared to Group 2 (40% +/- 7% versus 28% +/- 15% counts/pixel, p < 0.05) while left ventricular free wall uptake was similar. The ratio of right to left ventricular 201Tl uptake was increased in Group 1 versus Group 2 (0.81% +/- 0.30% versus 0.49% +/- 0.18%, p < 0.05). At 4 hr, right ventricular free wall 201Tl clearance was comparable, 51% +/- 13% versus 51% +/- 18% in Groups 1 and 2, respectively. No patient had perfusion abnormalities. Right ventricular ejection fraction was inversely related to dipyridamole stress right ventricular 201Tl uptake, r = 0.49, p < 0.03, s.e.e. = 13.6. Right ventricular 201Tl uptake was directly related to right ventricular wall thickness (r = 0.56, p = 0.18, s.e.e. = 10.4). Therefore, patients with more severe right ventricular systolic dysfunction have greater 201Tl uptake after dipyridamole stress, suggesting increased myocardial mass and possibly blood flow in response to hypertrophy. Patients with the most marked hypertrophy have impairment of right ventricular systolic function, independent of ischemia. PMID- 8410285 TI - Nuclear regulation: toward a balanced perspective. PMID- 8410286 TI - Lag phase in solid gastric emptying: comparison of quantification by physiological and mathematical definitions. AB - Lag phase is considered an important parameter of solid gastric emptying studies. However, different methodologies with their own normal values and physiologic implications are advocated for lag phase measurements. We applied both physiologic and mathematic approaches to quantify lag phases from identical image data sets for direct comparison of these two approaches. Gastric emptying studies were performed on 22 patients using a standard solid meal to calculate the lag phase in each using three different methods: (1) visual analysis to determine time when activity first appeared in the duodenum (LagPh); (2) time-activity curves to determine time of 2% decrease from peak stomach activity (LagCu); and (3) a mathematical definition using the modified power exponential method (TLAG). In addition, time of peak antral activity (AntPk) was calculated. The values for LagPh and LagCu were very similar (mean: 14.6 versus 15.2 min) and correlated well with each other (r = 0.89). TLAG using the power exponential showed different values (mean: 34.7 min) and correlated well only with AntPk (mean: 35.1 min; r = 0.92). We conclude that LagPh and LagCu estimate the time of onset of gastric emptying; the time when the smaller particles in the meal (< 1-2 mm) begin to leave the stomach (onset of variable emptying phase). On the other hand, TLAG and AntPk estimate total trituration time (time for most of meal to be processed) and signal the beginning of the constant gastric emptying phase. PMID- 8410287 TI - Early treatment response in malignant lymphoma, as determined by planar fluorine 18-fluorodeoxyglucose scintigraphy. AB - Clinical oncology needs flexible techniques for routine monitoring of treatment response. We therefore compared planar 18F-fluorodeoxyglucose (FDG) with a conventional gamma-camera and a special collimator to 67Ga scintigraphy in 26 patients with malignant lymphoma during chemotherapy. The scintigraphic appearance of involved sites was essentially the same with both tracers: in patients eventually achieving complete remission, tracer distribution had normalized after two courses; high uptake reflected treatment failure; faint uptake was associated with variable outcome. For (re)staging, 67Ga may be preferable (higher contrast). To document the initial response, we performed FDG scintigraphy during the first course (n = 11). Effective treatment sharply reduced metabolic tumor activity within days and prior to volume response, whereas abnormal uptake persisted in treatment failure. Planar FDG scintigraphy may be a tool to assess the potentially prognostic initial response rate, preventing over-treatment and allowing a timely switch to more aggressive therapy. PMID- 8410288 TI - Comparison of fluorine-18-fluorodeoxyglucose and carbon-11-methionine in head and neck cancer. AB - The positron emission tomography (PET) tracer 2-18F-fluoro-2-deoxy-D-glucose (FDG) is the most widely used tracer in oncology. PET tracer. Another radiotracer, L-methyl-11C-Methionine (11C-methionine), also has been used successfully for PET imaging of brain and lung tumors, non-Hodgkin's lymphoma, breast cancer and head and neck cancer. This study compared FDG and 11C methionine as tumor-detecting agents in head and neck cancer. Prior to cancer therapy, fourteen patients underwent a PET study with FDG and one with 11C methionine. Nineteen of 21 malignant lesions that could be evaluated were visible with both tracers. Tracer uptake was measured as standardized uptake values (SUV) and Ki values according to Patlak et al. The mean SUV in FDG studies was 7.7 +/- 4.2 and in 11C-methionine studies 7.7 +/- 2.5, whereas the Ki values in 11C methionine studies (mean, 0.128 +/- 0.068 min-1) were always higher than in FDG studies (mean, 0.036 +/- 0.023 min-1). A good correlation was found between the SUVs (r = 0.79, p < 0.0001) and the Ki values (r = 0.82, p < 0.001) between the two tracers. Both FDG and 11C-methionine are effective in PET imaging of head and neck cancer, and the uptake rates of the tracers seem to be closely related. PMID- 8410289 TI - Dopaminergic D2 receptor SPECT imaging in Rett syndrome: increase of specific binding in striatum. AB - A dopamine deficiency has been implicated in Rett syndrome, a progressive encephalopathy in girls that involves movement, tonus and cognitive disorders. To test the hypothesis that striatal D2 receptors increase in number in early stages of the disease, we measured the binding potential of 123I-Iodolisuride, a specific D2 ligand, in eleven Rett children aged 4-15 yr (7.9 +/- 3.5 yr) (mean +/- s.d.) and eight control subjects aged 3.5-13 yr (8.1 +/- 3.8 yr) who exhibited other neurological disorders. Regional cerebral blood flow (rCBF) was also measured with SPECT using 133Xe. The binding potential for 123I-ILIS and D2 receptors was significantly higher in Rett (0.45) than in controls (0.23) (p < 0.01). An increase in 123I-ILIS binding due to increased rCBF in patients' striata was excluded. Our results are consistent with a higher density of D2 receptors in young patients suffering from Rett syndrome because of reduced dopaminergic neurotransmission. PMID- 8410290 TI - Intracellular metabolism of indium-111-DTPA-labeled receptor targeted proteins. AB - The mechanisms of hepatic and renal accumulation and retention of 111In-labeled proteins has been the subject of many investigations. Because the lysosome is a common intracellular destination for a variety of agents including antibodies and polypeptide hormones, we studied the in vitro and in vivo metabolism of 111In DTPA-labeled polypeptides using a series of glycoproteins that were concentrated within the lysosome by receptor mediated endocytosis. Indium-111-DTPA-labeled glycoproteins targeted to the mannose, asialoglycoprotein and mannose 6-phosphate receptors were studied in vitro using cell lines known to express these receptors and in vivo using Sprague-Dawley rats. Once internalized, the 111In label was released slowly with 60%-90% (depending on the cell type) of the activity remaining cell associated at 24 hr. Subcellular fractionation using Percoll gradients indicated that the activity remained within the lysosome. Following internalization of the 111In-DTPA-labeled glycoproteins, the label was rapidly converted to a low molecular weight species (estimated molecular weight < or = 1000 daltons). This conversion was not seen with 111In-DTPA-alpha-galactosidase. As a lysosomal enzyme, alpha-galactosidase is relatively resistant to proteolysis within the lysosome. These results suggest that following internalization, 111In DTPA-polypeptides are delivered to the lysosome where the polypeptide backbone can be degraded to yield 111In-DTPA-amino acid(s). These metabolites remain within the lysosome and are only slowly released from the cell. The model systems used in these studies can also be used to evaluate the intracellular metabolism of polypeptides labeled by other techniques. PMID- 8410291 TI - Bromine-76-metabromobenzylguanidine: a PET radiotracer for mapping sympathetic nerves of the heart. AB - Iodine-123-metaiodobenzylguanidine (MIBG) is used to qualitatively assess heart innervation with single-photon emission computed tomography (SPECT). This approach is clinically useful in the prognostic evaluation of congestive heart failure. To improve quantification of uptake of the tracer using positron emission tomography (PET), we studied the characteristics of the bromoanalog of MIBG. Bromine-76-metabromobenzylguanidine (76Br-MBBG) was prepared from a heteroisotopic exchange between radioactive bromine atoms (noncarrier-added (76Br) BrNH4) and the cold iodine atoms of the precursor metaiodobenzylguanidine. Biodistribution was studied in rats and PET cardiac imaging performed in dogs. Myocardial uptake was high and prolonged in both species (mean half-life in dogs: 580 min). In rats, myocardial uptake was inhibited by desipramine by 64%, whereas after pretreatment with 6-hydroxydopamine uptake was reduced by 84%. In dogs pretreated with 6-hydroxydopamine or with desipramine, a steep washout of the tracer occurred (mean half-life: 136 min and 118 min, respectively). The non specific uptake plus the passive neuronal diffusion of the tracer could be estimated at about 25%-30% of the total fixation. In dogs, analysis of unchanged 76Br-MBBG in plasma showed that radiotracer metabolism was slow: 60 min after injection, 80% of the radioactivity was related to unchanged 76Br-MBBG. These preliminary findings suggest that 76Br-MBBG could be used to quantitatively assess adrenergic innervation in heart disease using PET. When combined with use of 11C-CGP 12177, cardiac adrenergic neurotransmission can be assessed. PMID- 8410292 TI - Scintigraphic evaluation of tenosynovial giant-cell tumor using technetium-99m(V) dimercaptosuccinic acid. AB - Technetium-99m(V) dimercaptosuccinic acid (DMSA) and 67Ga-citrate scintigraphy were performed in three patients with primary and recurrent tenosynovial giant cell tumor (one localized type and two diffuse type). In all cases, 99mTc(V)DMSA showed marked accumulation in all primary and recurrent tumors; however, 67Ga citrate showed no accumulation in any of the tumors. Technetium-99m(V)-DMSA scintigraphy was useful in detecting tenosynovial giant-cell tumor and in diagnosing recurrence of this tumor. PMID- 8410293 TI - Visualization of a recurrent carcinoid tumor and an occult distant metastasis by technetium-99m-sestamibi. AB - We present a case of atypical carcinoid that showed avid uptake of sestamib in a recurrent bronchial carcinoid tumor and a solitary, unsuspected, occult, bony metastatic lesion in the distal femur. As carcinoid tumors are known to be vascular, we suspect that its avidity for sestamibi is secondary to increased blood flow, although other factors such as the transmembrane potentials of plasma and mitochondrial membranes and relative number of mitochondria present in the cells of this carcinoid tumor, may have also played a role. PMID- 8410294 TI - A scanning line source for simultaneous emission and transmission measurements in SPECT. AB - A scanning collimated line source for simultaneously acquiring emission and transmission data from a gamma camera has been developed. The line source is microprocessor-controlled and incorporates hardware to electronically window the spatial gamma camera signals in order to separate the emission signals of the subject from transmission signals from the line source. The device improves upon the previously described emission-transmission scanning technique using a flood source in three ways: (1) it overcomes the limitation that the transmission radionuclide must have a lower energy than the emission radionuclide; (2) it provides narrow-beam (scatter free) attenuation measurements of the subject being examined; and (3) it reduces the radiation exposure to staff. Attenuation coefficients for an elliptocal water-filled phantom were measured to be mu = 0.15 +/- 0.01 cm-1. The technique has been validated in phantom and human studies using a range of radionuclide combinations and imaging geometries and gives equivalent results using separate and simultaneous acquisitions. PMID- 8410295 TI - Optimum sample times for single-injection, multisample renal clearance methods. AB - The best choice of sample times for measuring renal function in adults by single injection multisample plasma clearance methods was determined by Monte Carlo simulation, using a two-compartment model with parameters chosen to fit average values (from published clinical studies) for each of the three radiopharmaceuticals 99mTc-MAG3, 99mTc-DTPA and 131I-ortho-iodohippurate. Random errors were added and the simulated data were then fit to a two-exponential model using a weighted nonlinear curve fitting method. The calculated clearance values were compared with the original values to determine random and systematic errors for different selections of sample time for each radiopharmaceutical at various levels of renal function. The results show that for research-level accuracy with a GFR agent such as 99mTc-DTPA, plasma sampling must begin by 10 min after injection and continue at least 3 hr (in adults). With an ERPF agent such as 99mTc-MAG3 or 131I-OIH, sampling must begin by 5 min and continue for at least 90 min. Six logarithmically distributed samples are sufficient. PMID- 8410296 TI - Improved formulas for the estimation of renal depth in adults. AB - Commercial techniques are available to calculate effective renal plasma flow (ERPF) or glomerular filtration rate (GFR) based on the percent injected dose in the kidney 1-2 or 2-3 min post-injection; renal depth is estimated by the Tonnesen equations. Since the Tonnesen equations were derived from ultrasound measurements obtained at an oblique angle in sitting patients, we compared the renal depths obtained from the Tonnesen equations with the renal depth measured by computed tomography in supine patients, the most common position for radionuclide renography. The renal depth, height, weight, age and sex were determined for 126 patients undergoing CT scanning. Patients with obvious renal or abdominal pathology were excluded. The Tonnesen equations significantly underestimated renal depth. Using stepwise linear regression analysis, we derived a set of equations based on age, height and weight and applied these prospectively to a new set of 75 patients. In addition, a second set of equations were derived for the new data. There was no difference in the results for the two equations. We then pooled both studies and derived a combined set of equations: right renal depth (mm) = 153.1 weight/height + 0.22 age + 0.77 and left renal depth (mm) = 161.7 weight/height + 0.27 age - 9.4, where weight is in kilograms and height is in centimeters. The correlation coefficients were 0.81 and 0.83 for the right and left kidneys respectively with standard errors of the estimate of 10.2 and 10.1 mm. These equations provide a much better estimate of renal depth in the supine patient than the Tonnesen equations. PMID- 8410297 TI - Accurate local blood flow measurements with dynamic PET: fast determination of input function delay and dispersion by multilinear minimization. AB - Accurate determination of local blood flow in tissue using the Kety-Schmidt one compartment model for freely diffusible tracers requires knowledge of the true arterial input function in tissue. Because measured input functions are usually delayed and dispersed with respect to true influx, a correction of the experimental input function is necessary. We describe a technique that uses a fast multilinear least-squares minimization procedure to determine simultaneously the dispersion, the blood flow and the partition coefficient as a function of delay. In this approach, a few multilinear fits are sufficient to determine the complete set of parameters necessary to describe the data. Because of the high speed of the procedure, dispersion effects may be taken into account on a pixel by-pixel basis in calculating parametric images of blood flow and partition coefficient. The described procedure has been used at our institute for about 1 yr in more than 160 investigations and has proven well suited for routine use in a clinical environment. PMID- 8410298 TI - Physical performance evaluation of the Toshiba GCA-9300A triple-headed system. AB - The physical performance of the Toshiba GCA-9300A triple-headed SPECT system has been assessed. Using a water-filled cylinder containing 99mTc, the tomographic volume sensitivity was 33.8 and 34.8 kcps/(MBq/ml)/cm for the high-resolution, parallel-hole (HR-PH) collimator and the super high-resolution, lead fanbeam (SHR FB) collimator, respectively, excluding the rotation time(s) during scanning when data are not acquired. The tomographic spatial resolution at the center, in air, with 132 mm radius of rotation was 10.2 and 7.8 mm FWHM with the HR-PH and SHR-FB collimators, respectively; in water it was 11.0 and 7.8 mm. Reconstructed relative activity concentrations were accurate for both collimator sets if attenuation correction was used. With the SHR-FB collimators, the average peak-to valley ratio of five-line sources in water improved significantly when an asymmetric energy window was used. Using the three-dimensional Hoffman brain phantom which simulates a 4:1 grey matter-to-white matter ratio and the usual choice of acquisition and processing parameters for brain studies, the reconstructed grey matter-to-white matter ratio was only about 1.7 for total counts typically acquired in a 99mTc-HMPAO study (4.0 M counts) and only 2.3 for 40 M counts. There was a qualitative improvement with an asymmetric energy window. PMID- 8410299 TI - Doing well under pressure: dedicated SPECT cameras come of age. PMID- 8410300 TI - Rapidly converging iterative reconstruction algorithms in single-photon emission computed tomography. AB - Iterative reconstruction algorithms with markedly different convergence rates have been proposed in single-photon emission computed tomography (SPECT). Several new iterative reconstruction methods are described in this investigation. Differences between the methods include whether a ramp filter was used during backprojection, the type of backprojection weighting and whether camera and collimator blur were employed in the projection step. Simulated and real cylindrical phantoms with rod inserts were used to compare the properties of convergence and resolution following reconstruction by maximum likelihood (ML), iterative-Chang and the newly proposed reconstruction methods. Resolution was assessed after kernel-sieve regularization to achieve the same signal-to-noise ratio for all methods. Compared with maximum-likelihood reconstruction, methods employing a ramp converged much faster. One such method resulted in images with the same resolution and noise as ML, thus permitting termination of reconstruction at 14 iterations rather than the 1000 iterations required with ML. The major determinants of resolution were found to be use of an accurate model of the gamma camera imaging process in the projection step and inclusion of attenuation weighting and depth-dependent blur in the backprojection step. In summary, a new iterative reconstruction method was developed incorporating attenuation and blur and using a ramp filter that achieved results comparable to maximum-likelihood reconstruction in a fraction of the time. PMID- 8410301 TI - Time-dose-fractionation in radioimmunotherapy: implications for selecting radionuclides. AB - As currently practiced, the doses delivered to tumors in radioimmunotherapy are less than desirable primarily because of dose-limiting bone marrow toxicity, thus reducing the therapeutic efficacy of this modality. The biological effectiveness of internal radionuclide therapy depends on the total dose, the rate at which it is delivered, and the fractionation schedule of the radiolabeled antibodies administered. A new approach, based on time-dose-fractionation (TDF), which has been used in conventional radiotherapy, is advanced. This approach incorporates differences in dose rates, biological half-lives of the antibodies, physical half lives of the radionuclides employed and the total doses needed for a given biological effect. The TDF concept is illustrated with several relevant examples for radioimmunotherapy. Based on the TDF approach, it is proposed that under certain biological conditions radionuclides with physical half-lives that are 1-3 times the biological half-life of the radiolabeled antibodies in the tumor are more likely to deliver sterilization doses to tumors than the shorter-lived nuclides presently in use unless precluded by specific activity considerations. Several radionuclides that meet this criteria are suggested with 32P being the most promising among them. Finally, a practical method for treatment planning in radioimmunotherapy using TDF factors is recommended. PMID- 8410302 TI - Internal dosimetry using data derived from autoradiographs. AB - Cancer therapies based on administered radionuclides require accurate information on tumor dose. One of the major factors influencing the distribution of absorbed dose characteristics is the uniformity of the radiolabel distribution in tissue. To study the effect of nonuniformities, we used image analysis techniques to measure automatically the coordinates of autoradiographic grains (sources) and cell nuclei in cut sections from three different tumors, following treatment with radiolabeled antibodies. The spatial distribution data of sources and cell nuclei from these tumor sections were assessed and the pattern of energy deposition in the cell nuclei calculated, assuming that each autoradiograph grain corresponded to a source of the alpha emitter astatine-211 (211At) or the beta emitter yttrium 90 (90Y). The distribution of deposited energy obtained for the real grain distributions was compared to the distribution assuming a locally uniform source distribution, i.e., simulating grain count averaging as produced by a microdensitometric method within a 100 x 100 microns 2 frame size (frame averaging), and a uniform distribution across the entire section (section averaging). The results show first that when the grain distribution is uniform, the average dose within the section is an adequate estimate of the dose to the cell nuclei. Second, when the grain distribution is nonuniform, the distribution of doses to the cell nuclei is significantly less when calculations use the measured grain coordinates, or frame averaging, than when section averaging is used. Third, when the sources are located on or in the cells, both frame and section averaging produce underestimates of the dose to the cell nuclei. PMID- 8410303 TI - Gamma probe assisted ex vivo detection of small lymph node metastases following the administration of indium-111-labeled monoclonal antibodies to colorectal cancers. AB - This study evaluated the ability of ex vivo gamma-probe scanning to detect lymph node metastases in resected surgical specimens from primary colorectal carcinoma patients undergoing external scintigraphy following intravenous administration of 4.1-5.3 mCi of 111In-labeled anti CEA monoclonal antibodies. The ex vivo probe counting technique led to a twofold to fourfold increase in the number of detectable lymph nodes with the majority measuring 2-5 mm in diameter. Results indicate a potentially useful role for ex vivo probe counting in detection and mapping small (2-5 mm) lymph nodes metastases. PMID- 8410304 TI - On proselytism, retroversion and fiscal nihilism in nuclear cardiology. PMID- 8410305 TI - Human element integral to the future of nuclear medicine. PMID- 8410306 TI - No more Edsel mechanics needed in nuclear medicine. PMID- 8410307 TI - Nuclear medicine in Israel: independent, alive and well. PMID- 8410308 TI - Cultivating subspecialists for nuclear medicine. PMID- 8410309 TI - 1993 payment schedule threatens future of nuclear medicine specialty. PMID- 8410310 TI - The "cold hip" sign versus the avascular femoral head. PMID- 8410311 TI - Brookhaven National Laboratory and the BLIP upgrade. Brookhaven Linac Isotope Producer. PMID- 8410312 TI - Production of high purity fission molybdenum-99 in South Africa. PMID- 8410313 TI - The low occurrence of clinically significant ophthalmopathy after ablative treatment with radioactive iodine for Graves' hyperthyroidism. PMID- 8410314 TI - Is the Patlak graphical analysis method applicable to measurement of myocardial blood flow with nitrogen-13-ammonia? PMID- 8410315 TI - Rubidium-82 PET--essential or not? PMID- 8410316 TI - Interobserver variability in lung scintigraphy interpretation. PMID- 8410318 TI - Advantages and disadvantages of the SCANWriter report generating program. PMID- 8410317 TI - Complete nose closure and radioaerosol lung ventilation imaging. PMID- 8410320 TI - SNM restructuring plan passes board of trustees. PMID- 8410319 TI - New boss in town touts self-regulation. PMID- 8410321 TI - Nursing leadership and healthcare reform. Part I: Patient care delivery in a reformed healthcare system. PMID- 8410322 TI - The other side of shared governance. PMID- 8410323 TI - Reengineering. The work redesign--technology link. AB - Computerization in healthcare is a vital component of operational restructuring, but simply automating existing systems will not achieve the maximum benefit information technology has to offer. The authors present a discussion of information technology and healthcare, addressing a forecast of the predominant technologies in this decade, the impact of information technology in healthcare, and the use of reengineering to explore potential changes in the design of work made possible by information technology. The implications of reengineering in hospitals for nursing and nurse executives are explored. PMID- 8410324 TI - Nursing interventions classification. A comparison with the Omaha System and the Home Healthcare Classification. AB - Standardized languages for nursing practice are required to meet the needs of the profession and the patients we serve. The authors review and compare three classifications of nursing interventions: Nursing Interventions Classification (NIC), the Omaha System, and the Home Healthcare Classification (HHC). The information will help users make the best selection for their agency and client population. PMID- 8410325 TI - Organizing for communication and integration. AB - Using the departmental philosophy as its foundation, an integrated organizational model was designed for a nursing department. This patient-centered model was implemented to streamline operations, focus resources, and provide a professional practice environment for nursing staff. The "molecule" is used as a teaching tool and organizational framework, with applications ranging from orientation of new staff to management of daily operations. PMID- 8410326 TI - Women and nurse executives. Finally, some advantages. AB - How do chief nurse executives (CNEs) and chief executive officers (CEOs) compare on selected components of organizational socialization, and do differences exist between genders? To answer these questions, the author compared 127 male CEOs, 127 female CEOs, 232 female CNEs, and 117 male CNEs on their self-reported leadership styles, managerial values, and skills. The differences found between both genders and positions on all measures are largely advantageous to women and nurses in healthcare administration. PMID- 8410327 TI - The structured interview. An effective strategy for hiring. AB - Is your staff well prepared for interviewing job applicants? The potential for legal action and the cost incurred when turnover is high area two reasons administrators should help staff develop effective interviewing skills. The author proposes using the structured interview, which is more valid and reliable than other interview methods. The guidelines for conducting effective interviews will be useful to beginning managers and can serve as a refresher for experienced administrators. PMID- 8410328 TI - A community strategic planning process for elder services. AB - Dynamic changes in population, patient demographics, and healthcare financing mechanisms create shifting service priorities. Strategic planning has become a vitally important skill for nurse executives, who must extend their efforts and influence beyond traditional agency boundaries. The authors present an application of the strategic planning process as it was used to develop recommendations for eldercare services in one southwestern, urban community. PMID- 8410329 TI - Eliminating physical restraint use: implications for practice. PMID- 8410330 TI - The foundational principles of leadership. PMID- 8410331 TI - The nurse case manager in acute care settings. Job description and function. AB - Nurse administrators face many decisions about the emerging role of the nurse case manager (NCM). Who should be the NCM? What are the skills needed? How should the professional nurse function in the new role? The author reports the results of a study that helps answer these questions. PMID- 8410332 TI - Reflections on retention, recognition, and rewards. PMID- 8410333 TI - The chief nursing officer as board member. Risk management implications. PMID- 8410334 TI - Teams: group process, success, and barriers. AB - The business philosophy of total quality management is saturating the healthcare arena, and with it brings new meaning to the old concept of team. Nurses are not exempt from this concept, and will be expected to become effective team leaders and team members. The author reviews the literature on teams to define the concept of teams in the context of group process, and discuss effective team characteristics and barriers that often impede the success of teams. Whether the nurse carries the hat of educator, administrator, practitioner, or staff nurse, all will be part of a team within the changing healthcare environment. PMID- 8410335 TI - Implementing a clinically focused advancement system. One institution's experience. AB - Clinical ladders were designed to recognize, reward, and retain professional nurses who chose to remain in direct clinical practice. One institution implemented a clinically focused advancement system that addresses many of the pitfalls that have historically caused clinical ladders to fail. The authors describe unique features of the advancement system, along with the process for advancement, budgetary considerations, and transition to the new advancement system. PMID- 8410336 TI - Leader behavior in rural directors of nurses. AB - In several ways the scope of rural nursing practice differs from that of urban nursing. This study showed that rural rural directors of nursing are adjusting their leadership style to accommodate these differences. However, in the process of making these adjustments, the directors may be limiting their leader effectiveness. PMID- 8410337 TI - Enabling professional nursing staff to conduct research. PMID- 8410338 TI - Clinical practice guidelines. AB - With the emphasis on outcomes and quality improvement, nursing administrators need to encourage the integration of clinical practice guidelines into practice. Specific strategies for this integration are outlined, as are broad nursing implications for both patients and staff members. PMID- 8410339 TI - Management skills for directors of nursing. AB - What skills, tasks, and behaviors lead to effective management performance? The author reports a study in which directors of nursing were asked to assess their own skill performance and identify conflicts between management skills required of directors of nursing versus professional role expectations. Results provide a broader understanding of the director of nursing's role requirements and provide a validated base from which schools of nursing can plan curricula to prepare future nurse administrators. PMID- 8410340 TI - Length of stay: effect of interpleural catheters for cholecystectomy patients. PMID- 8410341 TI - Ambulatory nursing. The handmaiden/specialist dichotomy. AB - Will ambulatory nursing roles change sufficiently to cope with the healthcare demands of the next century or should they be taken over by administrative personnel? The authors analyze traditional and clinical nurse specialist ambulatory nursing activities and highlight the dichotomy between menial handmaiden and professional nursing activities that must be bridged if ambulatory nursing is to survive. PMID- 8410342 TI - Out of entitlement. A nursing organization's journey. AB - The authors describe a major reorganization that took place in an 850-bed academic teaching hospital in the southwestern United States. This reorganization facilitated a paradigm shift and the journey from a deeply entrenched attitude of "entitlement" to one of earning. PMID- 8410343 TI - Administrative rounds. A neglected art. AB - In making decisions, nurse administrators need diverse information from a variety of sources. One source of this information can be administrative rounds. Rounds not only offer opportunity for data collection and verification, problem identification and remediation, but they also provide an opportunity for coaching of staff and leadership visibility. The outcome of nursing work can be enhanced by nurse administrators seeing and being seen. PMID- 8410344 TI - Nurse executive practice in the Republic of South Africa. PMID- 8410345 TI - Excluding nursing administration education is shortsighted. PMID- 8410346 TI - The nurse-midwife as primary care provider. PMID- 8410347 TI - Preconception care. An opportunity to maximize health in pregnancy. AB - In 1990, the United States Public Health Service published Healthy People 2000: National Health Promotion and Disease Prevention Objectives. One of the objectives included in the family planning priority area and repeated in the maternal and infant health priority area is the following: "Increase to at least 60 percent the proportion of primary care providers who provide age-appropriate preconception care and counseling." Drawing on the guidelines proposed by the Public Health Service Expert Panel on the Content of Prenatal Care, this article describes the components of preconception care: 1) appropriate and ongoing risk assessment, 2) health promotion, and 3) medical and psychological interventions and follow-up. The organization of this article is based on a preconception class outline developed by the authors; recommendations included in the article are consistent with those of the Expert Panel. After discussing opportunities for providing preconception care, this article addresses: 1) helping women evaluate their psychological readiness; 2) evaluating physical readiness; 3) the examination and concerns of the father; 4) evaluating the need for genetic counseling; 5) creating a positive environment for conception; 6) discontinuing family planning methods and timing conception; and 7) choosing a provider and birth place. PMID- 8410348 TI - Eating and drinking in labor. A literature review. AB - This literature review questions the routine practice of denying food and fluids to women in labor. Fasting in labor, an established practice throughout the United States since the 1940s, is now under careful scrutiny. Many clinical practices, especially those that offer midwifery services, are currently instituting policies to allow and encourage eating and drinking in normal labor. To date, there have been no reported rises in maternal mortality with this policy change; neither have there been any reports of detrimental outcomes for mother or infant. PMID- 8410349 TI - Oligohydramnios. Literature review and case study. AB - Quantitative evaluation of amniotic fluid volume is now widely used to evaluate fetal status during pregnancy. A finding of decreased fluid volume raises management issues and requires that nurse-midwives arrange collaborative care. This article reviews the literature relevant to amniotic fluid volume and oligohydramnios. The significance to nurse-midwifery is discussed. Conservative management of oligohydramnios is described. A case involving oligohydramnios diagnosed prior to term is presented and discussed. PMID- 8410350 TI - ACNM-accredited nurse-midwifery education programs. Program information. PMID- 8410351 TI - Low-risk mothers. Oral intake and emesis in labor. AB - This study examined the pattern of oral intake and its impact on emesis and other complications in low-risk gravidas during labor. It is common clinical practice to restrict oral intake in most institutions. The historical bases for this restrictive practice are explored. Findings from this study indicate that when given a choice, all 106 women chose a variety of types and amounts of oral intake throughout all stages of labor. Over 80% of women who ate or drank during labor had no emesis. Of the 20 who did have any emesis, 40% (eight) vomited more than once. None of the women who vomited experienced poor outcomes. These data suggest that women who choose oral intake during labor are at relatively low risk for complications related to this intake. Based on a comprehensive review of the literature and these study results, practitioners should allow as much choice as is consistent with empirical knowledge and safe practice. PMID- 8410352 TI - Development of a single component of a performance evaluation protocol. The faculty impression score. AB - The issues of uniformity, consistency, and fairness in the measurement of behavioral skills constitute the ongoing debate of clinical evaluation. Evaluation tools, particularly those that propose to measure observed phenomena, have been judged to be limited in validity or reliability and are, therefore, more subjective than is desirable. Nurse-midwifery educators are challenged to identify or develop clinical and professional performance instruments, and to achieve interfaculty reliability in their application. In addition, nurse midwifery educational programs that make use of nonsalaried and/or off-site preceptors to contribute to student education must accept the dual challenges of development of clinical teaching skills and also the consistent application of the program's evaluation instruments and methods. The development of the "faculty impression score" as a supplemental, but critical, element within a behaviorally anchored, criterion-referenced evaluation protocol is presented. PMID- 8410353 TI - Nurse-midwifery service model in an academic environment. AB - The Nurse-Midwifery Division, with the full support of the Department of Obstetrics and Gynecology, and with committed partners in the College of Nursing, is proceeding forward with a three-part mission: nurse-midwifery practice, teaching, and research. Our practice sites are likely to remain varied but stable, with half of our clients being either Hispanic or Native American. Our commitment to nurse-midwifery education is solid, and our participation in the process will continue to take several forms, including the central one of clinical preceptorships. Our research agenda will grow. We hope our experience with GRAVI-DATA will contribute to the development of a national nurse-midwifery data base. Other future goals of our research program include collaboration in multisite clinical research projects, obtaining substantial external funding and recruiting doctorally prepared investigators. PMID- 8410354 TI - Speaking up and talking out. Barriers and obstacles to nurse-midwifery practice. AB - There are internal as well as external barriers to nurse-midwifery practice. We have let others define us. It is time to articulate a new vision for nurse midwifery that speaks to what midwifery does better than obstetrics. We need to claim our "partnership" model and argue for its integration into health policy for all childbearing women. PMID- 8410355 TI - Vitamin E is protective against iron toxicity and iron-induced hepatic vitamin E depletion in mice. AB - This study examined the effect of excess dietary iron on liver function, iron and vitamin E status and the protective activity of vitamin E. Consumption of excess dietary iron (3000, 5000, 8000 mg iron/kg/diet) compared with consumption of the control diet (120 mg iron/kg diet) by weanling male CD-1 mice for 7 wk resulted in accumulation of iron in liver, increased relative liver weights and a reduction in hepatic vitamin E stores. The concentration of vitamin E in the liver was negatively correlated with dietary iron concentration (r = 0.998). Weekly administration of vitamin E (20 mg/kg, subcutaneously) prevented iron induced liver damage without altering hepatic iron stores. Pretreatment of adult male CD-1 mice with a single subcutaneous dose of vitamin E (20 mg/kg body wt) 24 h prior to a lethal dose of iron (60 mg/kg, intraperitoneally) resulted in 100% protection. A similar dose of vitamin E given 5, 30 or 60 min (intravenously) after iron intoxication enhanced survival to 90, 70 and 80%, respectively, compared with the untreated control group. Vitamin E treatment 30 min after iron intoxication reduced mortality by 75% compared with intravenous treatment with 10 mg/kg of deferoxamine (Desferal). Data in this study indicate that vitamin E may be a useful antidote for iron toxicoses and that iron-induced depletion of vitamin E may play a role in the pathogenesis of iron toxicity. PMID- 8410356 TI - Enhanced insulin-dependent glucose utilization in iron-deficient veal calves. AB - Based on studies in Fe-deficient calves demonstrating enhanced blood lactate concentrations during treadmill exercise, the hypothesis was advanced that glucose metabolism is also disturbed at rest. Insulin-dependent glucose metabolism was therefore investigated in calves fed milk replacer containing 20 or 50 mg Fe/kg. Calves receiving only 20 mg Fe/kg of milk replacer developed moderate Fe deficiency anemia and had lower average daily gain than calves fed milk replacer containing 50 mg Fe/kg, but feed intake and feed refusals did not differ between groups. In Fe-deficient calves, insulin responses to glucose, based on intravenous and oral glucose tolerance and hyperglycemic clamp tests, were normal. Using hyperglycemic and euglycemic hyperinsulinemic clamps combined with [13C6]glucose infusions, glucose utilization and tissue sensitivity to insulin were greater in Fe-deficient calves than in Fe-adequate calves. Observed reductions in growth performance may be explained by decreased activity of Fe dependent enzymes, increased anaerobic glycolysis and lactate-glucose cycling, adaptations that are expected to be energy expensive. PMID- 8410357 TI - Docosahexaenoic acid is transferred through maternal diet to milk and to tissues of natural milk-fed piglets. AB - Docosahexaenoic acid (DHA) is incorporated in large amounts in structural lipids of the developing central nervous system. Milk DHA varies with maternal dietary DHA, but the effect of different intakes of DHA from milk on infant tissue fatty acids is unknown. The effect of milk high or low in DHA on the fatty acid composition of piglet brain, synaptic plasma membranes, retina, liver, plasma and RBC was studied. Pregnant sows were fed diets with 2.5 g/100 g vegetable oil until 4 d pre-partum and were then fed diets with 2.5 g/100 g soybean and canola oils or 4 g/100 g soybean oil plus 1 g/100 g fish oil to 15 d postpartum. Fish oil increased the milk DHA and eicosapentanoic acid from 0.1 to 1.5% and from 0.2 to 0.4% of fatty acids, respectively, but did not alter milk arachidonic acid. The level of DHA was significantly higher in plasma, liver and RBC phospholipids, brain and synaptic plasma membrane of 15-d-old piglets fed milk with high DHA compared with low DHA. Liver, plasma and RBC, but not brain or retina arachidonic acid, was significantly lower in piglets fed the high DHA milk compared with low DHA milk. Thus, differences in plasma, RBC and liver arachidonic acid and DHA of 15-d-old nursing piglets due to the maternal dietary fat were not accompanied by similar differences in central nervous system fatty acids. These studies show maternal DHA intake determines in part the infant plasma, RBC and liver phospholipid DHA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410358 TI - Starch bioavailability in arepas made from ordinary or high amylose corn: concentration and gastrointestinal fate of resistant starch in rats. AB - The purpose of the present investigation was to study the importance of the amylose/amylopectin ratio for the content and gastrointestinal fate of resistant starch in a realistic composite starchy food. Corn-based breads (arepas) from dent corn (25% amylose) and from high amylose corn (70% amylose) were used as test products. Resistant starch concentration was evaluated in vitro and in vivo using rats treated with an antibiotic drug (Nebacitin) to suppress hindgut fermentation. Experiments in rats with intact hindgut microflora allowed determination of resistant starch fermentability. The small intestinal digestibility of starch in dent corn arepas was close to 96% (total starch basis), whereas the starch in high amylose arepas was poorly digested (approximately 68%, total starch basis), as calculated from the fecal recovery of resistant starch in Nebacitin-treated animals. The main resistant starch fraction required solubilization in alkali to render it available to the analytical amylases (nonhydrated fraction). The total amount of resistant starch as well as the nonhydrated starch fraction delivered to the hindgut could be accurately predicted from analysis of starch remnants in an enzymatic gravimetric dietary fiber residue. Resistant starch present in dent corn arepas was fermented approximately 63%, whereas the fermentability of resistant starch from the high amylose product was remarkably low (< 11%). PMID- 8410359 TI - White sturgeon tissue fatty acid compositions are affected by dietary lipids. AB - Juvenile white sturgeon (Acipenser transmontanus) were fed eight isonitrogenous and isoenergetic purified diets for 9 wk to study their ability to utilize different dietary lipids. Each diet contained 15% of control oil mixture (corn oil-cold liver oil-lard, 1:1:1), corn oil, cod liver oil, lard, linseed oil, soybean oil, safflower oil or canola oil. No significant (P > 0.05) differences in percentage of body weight increase, feed efficiency or body composition were observed among sturgeon fed the different lipids. Tissue fatty acid compositions most sensitive to dietary lipids were those of muscle and liver, whereas brain fatty acid composition was the least sensitive. Results of this study indicate that it is possible to increase the levels of (n-3) polyunsaturated and highly unsaturated fatty acids in sturgeon muscle by feeding the fish lipids high in these fatty acids for 9 wk. PMID- 8410360 TI - Fat containing stearic acid increases fecal neutral steroid excretion and catabolism of low density lipoproteins without affecting plasma cholesterol concentration in hamsters fed a cholesterol-containing diet. AB - To examine the effect of different saturated fatty acids on the dietary cholesterol-induced elevation of serum cholesterol concentration and suppression of LDL catabolism, for 4 wk hamsters were fed purified diets containing 8% purified fats in which saturated fatty acids, lauric, myristic, palmitic and stearic acids were the sole variable. The dietary fat was composed of 50% saturated fatty acid, 30% oleic acid and 20% linoleic acid (polyunsaturated:saturated = 0.4). In hamsters fed the cholesterol-containing diet, fat containing stearic acid, compared with the fats containing other saturated fatty acids resulted in greater fractional catabolic rate of [125I] labeled homologous LDL, greater fecal excretion of neutral steroids and lower liver cholesterol concentration, although the elevation of serum cholesterol concentration due to consuming a cholesterol-containing diet was not ameliorated. Stearic acid fat resulted in greater excretion of fecal fatty acids and lower apparent absorption of the dietary fats in hamsters fed diets with and without cholesterol. In hamsters fed the cholesterol-free diets, type of dietary fat did not affect the fractional catabolic rate of LDL, although stearic acid fat resulted in greater fecal neutral steroid excretion and lower serum and liver cholesterol concentrations. These observations suggested that purified fat containing stearic acid results in lower plasma cholesterol concentration in hamsters via stimulation of neutral steroid excretion, but addition of cholesterol to the diets obscures this effect. PMID- 8410361 TI - Dietary fish oil alters rat milk composition and liver and brain fatty acid composition of fetal and neonatal rats. AB - This study was performed to determine the effects of dietary fish oil rich in docosahexaenoic acid [22:6(n-3)] on the milk composition of maternal rats and tissue fatty acid composition of fetal and neonatal rats. The fatty acid composition of dam's milk was affected by the dietary fat. The level of 22: 6(n 3) in the milk of dams fed the fish oil-containing diet was severalfold higher than that of dams fed the diet that did not contain fish oil, and the levels of 22: 6(n-3) in the phospholipids of the livers and brains of fetuses and pups of the fish oil-fed group were higher than those in fetuses and pups of the group that did not receive fish oil. These findings suggest that a high level of 22:6(n 3) in the livers and brains of fetal and neonatal rats can be achieved by intake of fish oil by the dams. PMID- 8410362 TI - Dietary soybean protein compared with casein damages colonic epithelium and stimulates colonic epithelial proliferation in rats. AB - Recently we showed that soybean protein compared with casein stimulates the fecal excretion of endogenous magnesium. In the present study we investigated whether this differential effect of soybean protein is due to intestinal epithelial cell damage and thus results in a compensating increase in proliferation of epithelial cells. Two groups of six rats were fed purified high fat, low calcium diets, differing only in the type of protein. When compared with casein feeding, soybean protein feeding stimulated the fecal excretion of magnesium, fat and fatty acids, but had no effect on the excretion of bile acids. In fecal water, the concentration of bile acids was lower when soybean protein was fed. In contrast, free fatty acid concentration, as well as luminal cytolytic activity, was higher in fecal water of rats fed soybean protein. Furthermore, epithelial cell damage and proliferation of colonic epithelium (measured as in vivo incorporation of tritiated thymidine into DNA) were greater in rats fed soybean protein. The stimulation of colonic proliferation by soybean protein is consistent with the observed increase in luminal cytolytic activity and epithelial cell damage. We conclude that the stimulatory effect of soybean protein on endogenous magnesium excretion is due to a soybean protein-specific damage of colonic epithelial cells, which results in a compensatory epithelial cell hyperproliferation. PMID- 8410363 TI - Dietary energy source and density modulate the expression of immunologic stress in chicks. AB - To determine how dietary energy level and source influence feed intake, growth and energy partitioning drug immunologic stress, growing chicks were fed diets based on cornstarch and casein with varying energy densities and injected every other day for 6 d with either saline (control), Salmonella typhimurium lipopolysaccharide or heat-killed Staphylococcus aureus. Salmonella typhimurium lipopolysaccharide decreased growth and feed consumption at low energy densities. When the dietary energy density was increased above 13.4 kJ/g using cornstarch, but not corn oil, the growth depressing effect of immunogens was eliminated. Immunologically stressed chicks had a greater proportion of gain in visceral organs and less in the carcass, regardless of the nutrient density of the diet. Immunologic stress decreased intake of metabolizable energy of chicks fed a diet with low nutrient density and increased it for those fed a diet with high nutrient density. Chicks injected with S. typhimurium lipopolysaccharide lost more energy as heat than controls when differences in metabolizable energy intakes were accounted for and modified their preference between two diets differing in metabolizable energy density and fat content as a result of the challenge. Control chicks selected between the 11.7 and 14.2 kJ/g diets to obtain an energy density of 13.2 kJ/g compared with 12.5 kJ/g in the S. typhimurium lipopolysaccharide-challenged chicks. The S. typhimurium lipopolysaccharide challenged chicks consumed similar amounts of the low energy diet but decreased intake of the high energy diet. PMID- 8410364 TI - Dietary native resistant starch but not retrograded resistant starch raises magnesium and calcium absorption in rats. AB - The effects on calcium and magnesium absorption of dietary native and retrograded cornstarch were studied in rats. Uncooked high amylose starch granules (35% of total glucose equivalents as enzyme-resistant starch) and cooked and cooled (-20 degrees C) high amylose starch (24% of total glucose equivalents as retrograded resistant starch) were used as test starches, and cooked normal starch (3% of total glucose equivalents as resistant starch) was used as control starch. Native vs. control starch raised the amount of polymerized glucose in ileum, but not in feces. Retrograded starch produced more polymerized glucose than control starch in both ileum and feces. When compared with control starch, ileal pH was significantly lowered by native starch and tended to be raised by retrograded starch. Cecal pH was lowered by the two preparations rich in resistant starch. Apparent absorption of calcium and magnesium was raised by native starch but not by retrograded resistant starch. Calcium concentrations in the liquid phase of the ileum tended to be elevated by native starch but were significantly lowered by retrograded starch relative to control starch. Magnesium and calcium concentrations in liquid cecal contents tended to be raised with native starch; they were unchanged with retrograded starch. It is suggested that native resistant starch raised calcium and magnesium absorption because it tended to enhance the solubility of these minerals in ileal and cecal digesta. PMID- 8410365 TI - Relationships between viscosity of hydroxypropyl methylcellulose and plasma cholesterol in hamsters. AB - Dietary high viscosity hydroxypropyl methylcellulose (HPMC) lowered plasma and liver cholesterol concentrations in cholesterol-fed hamsters. To determine the level of viscosity needed to effect a significant reduction in total plasma cholesterol, hamsters were fed for 3 wk diets containing 0.12% cholesterol and either 4% cellulose or one of four preparations of HPMC that varied in viscosity between 14 and 1698 centipoise (cP), as estimated in vitro. Blood was collected for plasma cholesterol determination, and intestinal contents were obtained by finger-stripping of the excised small intestine. Contents were centrifuged and the supernatant (ex vivo) viscosity determined. In vitro and ex vivo viscosities were correlated (R2 = 0.96). Plasma cholesterol concentrations declined as in vitro or ex vivo viscosity increased. Maximal plasma cholesterol reduction occurred at an ex vivo viscosity of approximately 150 cP. There was a linear relationship between plasma cholesterol and the logarithm of ex vivo viscosity (R2 = 0.98). Our results suggest that materials that increase the viscosity of intestinal contents can be effective in reducing plasma cholesterol and that only moderate increases in viscosity are necessary to achieve this effect. PMID- 8410366 TI - Hepatic denervation alters the transition from the fed to the food-deprived state in conscious dogs. AB - The hepatic nerves can modulate hepatic glycogenolysis and glycogenesis and thus might be expected to be involved in the response of the animal to the transition from the fed to the food-deprived state. Therefore the arterial concentrations and net hepatic balance of glucose and its metabolites, as well as the hepatic glycogen concentrations, were compared in hepatic-innervated and -denervated dogs 18, 24 and 42 h after their usual daily meal. Arterial concentrations of glucose, alanine, lactate and glycerol; net hepatic balances of glucose, alanine and glycerol; and glycogen concentrations were similar in hepatic-innervated and denervated dogs at each time investigated. Net hepatic balances of lactate (with negative values indicating uptake) in hepatic-innervated and -denervated dogs, respectively, were: 18 h, 4.1 +/- 4.3 vs. -4.3 +/- 3.6 mumol.kg-1 x min-1; 24 h, 4.8 +/- 3.6 vs. -6.7 +/- 1.7 mumol.kg-1 x min-1 (P < 0.05); 42 h, -7.0 +/- 2.0 vs. -6.8 +/- 1.0 mumol.kg-1 x min-1. Based on changes in net hepatic lactate balance, the denervated liver responds more rapidly to food deprivation than the innervated liver, but the metabolic state of the liver appears similar by 42 h after a meal. PMID- 8410367 TI - Carbohydrate utilization by tilapia (Oreochromis niloticus x O. aureus) as influenced by different chromium sources. AB - An experiment was conducted to study the influence of three different forms of chromium (CrCl3.6H2O, Na2CrO4.4H2O and Cr2O3) on the utilization of two carbohydrates (glucose and cornstarch) by juvenile hybrid tilapia (Oreochromis niloticus x O. aureus). Average initial body weight of the fish was 1.13 +/- 0.02 g. Fish were fed 5% body wt/d. Significantly (P < 0.05) greater body weight gain, food intake, protein retention, energy retention and body lipid concentration were observed in fish fed the starch diet than in those fed the glucose diet. Fish fed the glucose diet supplemented with any type of chromium had significantly greater weight gain than those fed the glucose diet without chromium supplementation. Fish fed the glucose diet supplemented with Cr2O3 had greater weight gain, food intake, protein retention, energy retention and body lipid concentration than those fed the unsupplemented glucose diet or the glucose diet supplemented with CrCl3.6H2O or Na2CrO4.4H2O. Delayed plasma glucose peak time was observed in tilapia fed the glucose diet supplemented with any type of chromium. Chromium supplementation generally lowered the glucose-6-phosphatase activity in tilapia. Phosphofructokinase activity was significantly higher in fish fed the glucose diet supplemented with Cr2O3 than in the other glucose-fed groups. These data suggest that chromium supplementation improved glucose utilization by tilapia and that Cr2O3 supplementation was markedly more effective than other chromium forms. PMID- 8410368 TI - Augmentation of protein synthesis and degradation by poor dietary amino acid balance in European sea bass (Dicentrarchus labrax). AB - Sea bass fry were fed a fixed ration of one of six isonitrogenous diets differing in essential amino acid balance or physical and chemical state of the protein source (Hydrolysate vs. intact protein) to induce different growth rates. The reference diet was based on fish meal, whereas the other diets contained fish protein hydrolysate, greaves meal (i.e., defatted collagen meal) or hydrolyzed feather meal added at 30 or 50% of crude protein at the expense of fish meal protein. Digestibility as well as fractional rate of whole-body protein synthesis was assessed. Whole-body protein synthesis was determined for each group of fish using a single injection of flooding dose of tritiated phenylalanine. Protein digestibility of the diets varied only by 5.5%. Specific growth rate and fractional protein specific growth rate, i.e., fractional protein accretion, were higher in fish fed the reference diet than in those fed the diets in which 50% of fish meal protein had been replaced by greaves or hydrolyzed feather meal protein. Compared with the reference group, whole-body protein synthesis was higher in fish fed these latter diets as well as in those fed the diet containing 30% greaves meal protein. The fractional protein accretion to fractional protein synthesis ratio, i.e., the efficiency of protein deposition, was lower in fish fed poorer dietary amino acid balance than in the reference group. The substitution of fish protein hydrolysate for intact fish protein led to a similar, though less pronounced phenomenon: nonsignificant increase in protein synthesis accompanied by significant increase in protein degradation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410370 TI - The Directory of Graduate Programs in Nutritional Sciences. PMID- 8410369 TI - Glycerophosphocholine and phosphocholine are the major choline metabolites in rat milk. AB - Choline is a constituent of cell membranes, surfactant and acetylcholine and is also a major source of methyl groups for the regeneration of methionine from homocysteine. Previous analyses of rat, human and bovine milk measured only choline, phosphatidylcholine and sphingomyelin. Choline-containing compounds in milk from rats lactating for 15 d were measured by HPLC and gas chromatograph mass spectrometry. In addition to the previously reported choline metabolites, substantial concentrations of glycerophosphocholine (3.7 mmol/L) and phosphocholine (653 mumol/L) were also detected. At 1 h after oral administration of [methyl-14C]choline to lactating rats, the major labeled metabolites were phosphocholine (91% of label in milk) and betaine (9%). Twenty-four hours after the dose, glycerophosphocholine was the major labeled metabolite (69% of label in milk). Rat mammary epithelial cells, in primary culture, synthesized and secreted phosphatidylcholine, phosphocholine, glycerophosphocholine and betaine. Thus, the mammary gland was able to synthesize the choline metabolites found in milk, but these metabolites may not be derived exclusively from uptake from maternal blood. We have established that the total choline concentration in rat milk is sevenfold higher than previously reported, with > 80% present as glycerophosphocholine and phosphocholine. PMID- 8410371 TI - Gross and separative determination of 1 alpha,25-dihydroxyvitamin D2 and D3 in plasma using calf thymus receptor. AB - Determination of 1 alpha,25-dihydroxyvitamin D [1,25(OH)2D, gross amounts of 1,25(OH)2D2 and 1,25(OH)2D3] and separative determination of 1,25(OH)2D2 and 1,25(OH)2D3 in plasma using calf thymus receptor have been investigated. A lipid extract from 1 ml of plasma is applied to a Bond Elut C18OH column and an eluate corresponding to 1,25-(OH)2D including both 1,25(OH)2D2 and 1,25(OH)2D3 is applied to calf thymus receptor to assay a gross amount of the two compounds. On the other hand, when separative assay of the two compounds is performed, the 1,25(OH)2D eluate obtained from the Bond Elut C18OH column is further applied to HPLC using a Zorbax SIL column with 5% isopropanol in methylene chloride as a developing solvent to separate the two compounds from one another. The separated eluates are independently applied to the receptor to assay the two compounds. Since less amounts of unknown components non-specifically bound to interfering concomitants besides 1,25(OH)2D exist in the calf thymus receptor, complicated purification steps to eliminate the concomitants are unnecessary. The detection limit by this method is 1.25 pg/tube which is sensitive enough for a routine method to assay 1,25(OH)2D in plasma. PMID- 8410372 TI - Importance of the nucleotide loop moiety coordinated to the cobalt atom of adenosylcobalamin for coenzymic function in the diol dehydrase reaction. AB - Three analogs of adenosylcobalamin were synthesized and their coenzymic properties in the diol dehydrase reaction were studied. Neither adenosylcobinamide nor adenosylcobinamide phosphate was active as coenzyme and showed very low affinity for apoenzyme, irrespective of the presence of nucleotide loop fragments, such as 5,6-dimethylbenzimidazole, alpha-D-ribazole, or alpha-D-ribazole-3'-phosphate. The coordination of pyridine to the cobalt atom neither confers the coenzymic function upon adenosylcobinamide nor strengthens the inhibitory effect of cyanoaquacobinamide and methylcobinamide significantly. The analog of adenosylcobalamin in which the N-3 position of 5,6-dimethylbenz imidazole is methylated was also not active as coenzyme and showed very low affinity for apoenzyme. Since 3,5,6-trimethylbenzimidazole in this analog is no longer coordinated to the cobalt atom, these results show that at least a part of the nucleotide loop moiety coordinated to the cobalt atom of adenosylcobalamin is essential for tight binding to the apoenzyme and therefore for manifestation of coenzymic function. PMID- 8410373 TI - Relationship between total serum cholesterol level and nutritional and physical status in Nepalese rural people. AB - To investigate the nutritional condition in a hilly village (Kotyang) and a suburban village (Bhadrakali) in Nepal and to clarify the possible cause of the difference in total serum cholesterol level between the two groups of villagers habitually eating low fat diets, we carried out a nutrition survey using the 24-h recall method and blood sampling in 403 subjects (204 men and 199 women) in the hilly village and 466 (244 men and 222 women) in the suburban village. Total serum cholesterol was statistically significantly lower in the hilly villagers than in the suburban villagers for both sexes, but HDL-cholesterol was not. In both villages, 82% of the total energy was taken from carbohydrate, 7-8% from fat and 10% from protein. Energy, protein, fiber, potassium, magnesium, monounsaturated fatty acid, polyunsaturated fatty acid, and vitamin A in the hilly villagers were significantly higher than those in the suburban villagers. Total serum cholesterol was significantly associated with age and body fat percentage, suggesting that total serum cholesterol level was not directly associated with total fat intake in these Nepalese people. PMID- 8410374 TI - Altered lipid metabolism during enteral or parenteral nutrition in rats: comparison with oral feeding. AB - To investigate the effects of different routes of alimentation on lipid metabolism, lipid-free nutrients with the same amount of energy and composition were continuously administered via the oral cavity (oral group), directly to the stomach (enteral group), or into the superior caval vein (parenteral group) of unrestrained rats. The body weight gain 1 week after continuous nutrition was greater in enteral and parenteral groups than in the orally-fed group. In comparison with the orally-fed group, the enterally-fed group had significantly greater liver and retroperitoneal adipose tissue weights and hepatic lipid content, whereas the parenterally-fed group produced similar changes to those in the enteral group without significant accumulation of hepatic lipid. The rate of fatty acid synthesis after 1 week of alimentation was 3-fold higher in the liver in enterally-fed group, and approximately 10-fold higher in white adipose tissue in both enterally- and parenterally-fed groups than in orally-fed group. Plasma concentrations of catecholamines after 6 h were significantly higher in the orally-fed group than in either the enteral or parenteral group. However, plasma insulin concentrations were not significantly different among the three groups. The results indicate that lipid synthesis and its deposition in the liver and adipose tissue are greatly influenced by the route of alimentation, possibly owing to difference in the early neuro-hormonal responses to different routes of alimentation. PMID- 8410375 TI - Effects of vegetable oils and C18-unsaturated fatty acids on plasma ethanol levels and gastric emptying in ethanol-administered rats. AB - The effects of vegetable oils (soybean oil and coconut oil) and C18-unsaturated fatty acids (oleic acid, C18:1; linoleic acid, C18:2; linolenic acid, C18:3) on plasma ethanol levels in male rats (6 weeks old) were investigated. Vegetable oils decreased and delayed the peak of plasma ethanol concentration: a dose response to vegetable oils was observed in the concentration and time to maximum concentration of plasma ethanol but no change in disappearance time. These phenomena were observed in two conditions: 1) oral administration of vegetable oils before oral intubation of ethanol and 2) simultaneous oral administration of vegetable oils and ethanol. Similar responses were obtained in three C18 unsaturated fatty acids. No changes in hepatic alcohol and aldehyde dehydrogenase isozyme (high Km and low Km) activities were observed. The remaining rate of ethanol in stomach was significantly higher with administration of vegetable oils or linoleic acid. A high negative correlation between the maximum plasma ethanol concentration and the remaining rate of ethanol in stomach was found. These results suggest that the slowing of the gastric emptying is a major mechanism for the decreasing and delaying effects on plasma ethanol levels by vegetable oils. The present paper also suggests that fatty acids may participate in the decreasing and delaying actions on the peak of plasma ethanol concentration by vegetable oils. PMID- 8410376 TI - Teratogenic effects of triethylene tetramine dihydrochloride on the mouse brain. AB - The teratogenic effects of triethylene tetramine dihydrochloride (Trien-2HCl) on fetal mouse brain were studied on gestational day 19. Trien-2HCl was given throughout pregnancy at levels of 0 (control), 3,000, 6,000, or 12,000 mg/liter as drinking water, ad libitum. Mean litter size and live fetus per dam at birth were not significantly different among the four groups. The frequency of gross brain abnormalities in live fetus at birth such as hemorrhages, delayed ossification in cranium, hydrocephaly, exencephaly, and microcephaly increased with increasing levels of the drug. Microscopically, disorganization of neuronal cell layers, spongiform changes in white matter, and reduced myelin development were noted in the coronally sectioned cerebrum from Trien-2HCl-treated fetus. These abnormal findings increased dose-dependently in regard to the extent and severity at the levels of 6,000 and 12,000 mg/liter. No such changes were observed in the cerebrum of controls. These results suggest that microscopic changes in fetal brain caused by Trien-2HCl may be in part similar to those in brindled mutant mouse. Special attention should be paid to the developing fetal brain when Trien-2HCl is used during pregnancy. PMID- 8410377 TI - Changes in zinc and copper concentrations in breast milk and blood of Japanese women during lactation. AB - To evaluate the changes in zinc and copper concentrations in breast milk and maternal blood during lactation, milk and blood samples were obtained from 80 lactating women during the period between 2 and 201 days of lactation. Zinc and copper concentrations were measured by atomic absorption spectrophotometry. Breast milk zinc and copper concentrations markedly decreased during the first few weeks of lactation and gradually declined for the remaining period. Mean values of milk zinc and copper levels were 1.76 and 0.29 micrograms/ml, respectively, between 15 and 84 days after parturition and were 0.76 and 0.19 microgram/ml between 85 and 201 days of lactation. Calculated daily intakes of these minerals for infants from breast milk were markedly lower than those of US Recommended Dietary Allowances. Plasma zinc levels of lactating mothers increased as lactation progressed, whereas erythrocyte zinc and plasma copper concentrations decreased. Plasma zinc and copper and erythrocyte zinc values returned to normal approximately three months after parturition. PMID- 8410378 TI - Immunoelectron microscopic localization of sucrase-isomaltase in rat small intestine. AB - The localization of rat small intestinal microvillous sucrase-isomaltase was studied by immunoelectron microscopy to investigate its intracellular transport to the microvillous membrane. At the same time, the usefulness of a rapid embedding method of tissues in Lowicryl K4M for immunocytochemistry of sucrase isomaltase was examined. Sucrase-isomaltase was present not only in the microvillous membrane, but also in the apical vesicles and the apical plasma membrane invaginations. Negligible labeling was observed in the other portions of the absorptive cells. These findings suggest that the final step of intracellular transport of sucrase-isomaltase to the microvillous membrane is via smooth apical vesicles. The rapid immunoelectron microscopic method adopted in this study seemed to be a useful technique for the study of the intracellular localization of sucrase-isomaltase. PMID- 8410379 TI - Increase of renal 25-hydroxyvitamin D3-24-hydroxylase activity and its messenger ribonucleic acid level in 1 alpha-hydroxyvitamin D3-administered rats: possibility of the presence of two forms of 24-hydroxylase. AB - Messenger ribonucleic acid (mRNA) levels and activities of renal 25 hydroxyvitamin D3-24-hydroxylase (24-OHase) were determined in 3 groups of rats: vitamin D-deficient, normal, and 1 alpha-hydroxyvitamin D3 (1 alpha OHD3) administered rats. To measure renal 24-OHase mRNA, a DNA probe complementary to the reported sequence of the recently cloned P450 component was employed. The DNA probe hybridized with 24-OHase mRNA. Northern blot analysis indicated that the size of the transcript was approximately 3.4 kb, which was similar in size to that of the previous report. Oral administration of 1 alpha OHD3 (2 micrograms/kg bw/day x 7 days) markedly increased the plasma concentration of 1,25 dihydroxyvitamin D (1,25(OH)2D), renal 24-OHase activity and the renal 24-OHase mRNA level. However, the levels of 24-OHase mRNA were undetectable in vitamin D deficient and normal rat kidneys, while a significant amount of 24-OHase activity was observed in the normal rat kidney. In vitamin D-deficient rats, renal 25 hydroxyvitamin D3-1 alpha-hydroxylase activity was markedly elevated and 24-OHase activity was completely abolished. When an oligonucleotide probe complementary to coding sequence of heme binding region was used for dot blot analysis, the mRNA was clearly detected in normal rat kidney. Furthermore, activity of the enzyme was attenuated by human parathyroid hormone (1-34) but the mRNA abundance did not change with 2 nmol (every 4 h, 5 times) of hormone treatment in 1 alpha OHD3 dosed thyroparathyroidectomized rats. The present study demonstrates that induction of 24-OHase activity by 1,25(OH)2D3 occurs at least in large part through increase of the gene expression in the kidney. Furthermore, these findings suggest that 2 forms (inducible and constitutive) of 24-OHase exist in rat kidney, and that the previously cloned P450 component is the inducible form of 24-OHase. PMID- 8410380 TI - Partnership in perinatal care. PMID- 8410381 TI - Improving survival of the very premature infant. AB - Knowledge of current data on survival and morbidity for premature infants is essential for perinatal decision making and accurate counseling of families. Survival and severe morbidity were reviewed for liveborn infants of less than 28 weeks' gestation. Between July 1989 and August 1991, in a cohort of 93 infants with a mean birth weight of 887 gm, survival until hospital discharge to home was 73%. Twenty-eight percent of survivors had either grade 3 or 4 intraventricular hemorrhage or an oxygen requirement at hospital discharge. In the subset of infants of less than 26 weeks' gestation, survival has significantly improved without a rise in morbidity when the data from the study period were compared with data from 1986 through 1988. Survival and morbidity were similar between black and white infants and between girls and boys. These data, along with reports from other tertiary centers, indicate a generalized improvement in the outcome of infants of very low birth weight. PMID- 8410382 TI - Calcium and phosphorus supplementation after initial hospital discharge in breast fed infants of less than 1800 grams birth weight. AB - This study evaluated whether calcium and phosphorus supplementation after initial hospital discharge was advisable in infants of < 1800 gm birth weight who were being breast fed. Twenty-seven infants (15 without any illness affecting nutritional intake and 12 with medical illness) received breast milk plus a liquid human milk fortifier mixed 1:1 and 400 IU vitamin D daily during initial hospitalization. At discharge, 12 infants (6 without and 6 with previous illness) were randomly assigned to receive calcium and phosphorus supplementation, and 15 infants (9 without illness and 6 with previous illness) received no mineral supplementation. A third group of seven healthy infants received a formula for premature infants during initial hospitalization and a standard cow's milk formula (20 calories per ounce) after discharge. The mean plasma calcium, phosphorus, and alkaline phosphatase levels did not differ among the three groups at study entry. Eight weeks after discharge, eight infants (four without illness and four with illness) had hypophosphatemia < 4.5 mg/dl. All were breast fed, and seven of eight had not received posthospitalization calcium and phosphorus supplementation. The incidence of hypophosphatemia in infants with or without illness was significantly greater in infants who did not receive supplementation (p = 0.038). These data indicate that calcium, phosphorus, and vitamin D supplementation may be necessary in approximately 50% of breast-fed infants of < 1800 gm birth weight after hospital discharge. It is recommended that serum calcium, phosphorus, and alkaline phosphatase be measured 4 to 8 weeks after discharge to identify those infants who require supplementation. PMID- 8410383 TI - Experience of risk for pregnant black women. AB - This qualitative study explores how risk is determined by black women during pregnancy. A total of 17 women were assessed for risk; 7 were at risk for preterm birth, and 10 were not at risk. These women were interviewed by using an open ended interview guide. The transcribed interviews were analyzed by using grounded theory. Risk was defined by the provider's assessment of a mathematical probability of the occurrence of preterm labor according to the risk screening tool developed by Creasy. Risk was also defined by black women on the basis of their experience of problematic change, on the counsel of other black women, and on the assessment of the provider. All of the women perceived their pregnancy as a normal process. For some, that perception never changed; for others, it did change. The change was precipitated by the occurrence of an unexpected event, indicated in this study as a critical moment. The critical moment is a dynamic interplay among biophysical changes, patterns of social interaction, and intersubjective reflection. These findings emphasize the black woman's reliance on herself in problematic situations and the significant role of sharing between black women in perpetuating their culture's normative expectations concerning pregnancy. PMID- 8410384 TI - Sudden death in neonate with staphylococcal endocarditis. AB - A 27-week-old girl, 936 gm, with initial diagnosis of birth asphyxia resulting from prolapse of the umbilical cord, respiratory distress syndrome, and suspected neonatal sepsis received antibiotics for the first 7 days of life. On day 24, evidence developed of acute Staphylococcus aureus endocarditis of the anterior leaflet of the mitral valve, and she died 4 days later. We believe that a rapid enlargement of the endocardial vegetation caused acute occlusion of the small mitral valve and was the cause of sudden death in this infant. PMID- 8410385 TI - Regionalization of infant transports: the southern California experience and its implications. I: Referral pattern. AB - The development and implementation of neonatal intensive care services have been essential components of perinatal regionalization during the past two decades. The transport of critically ill infants to regional neonatal intensive care units has played an important role in improving neonatal outcome. This article presents a profile of Southern California's 10-year infant transport experience (1979 through 1988) in terms of the following indicators: transport volume, cross county referrals, distance travelled, referral pattern, and birth weight pattern. Findings point to the necessity of focusing attention on several critical issues confronting Southern California's neonatal services in the 1990s. They include adequacy of tertiary or intermediate bed capacity in neonatal intensive care units; appropriateness of existing infant referral practices; impact of maternal fetal transports; availability of financing resources; and overall differences in morbidity and mortality rates between transported and nontransported infants in subpopulations. PMID- 8410386 TI - Indomethacin and recurrent ileal perforations in a preterm infant. AB - This article reports a case of a very low birth weight infant who was given intravenous indomethacin for a symptomatic patent ductus arteriosus and subsequently had two isolated ileal perforations several days apart. Spontaneous or indomethacin-related ileal perforation appears to constitute a separate clinical and pathologic entity, different from necrotizing enterocolitis, with a benign clinical picture and good prognosis when promptly recognized. PMID- 8410387 TI - Body image and pregnancy: bridging the mind-body connection. A guide for health care professionals. PMID- 8410388 TI - Continuing education: the newborn nurse. PMID- 8410389 TI - Home apnea monitoring and risk factors for poor family functioning. AB - As part of a comprehensive study on the impact of home apnea monitoring on family functioning, we undertook an in-depth analysis of risk factors for poor family functioning in 93 families of monitored infants. A multistep correlation and regression analysis was used to examine the ability of 16 demographic, infant, family, and monitor-related variables to predict poor functioning in each of 12 different aspects of family life. Of note was the fact that monitor-related variables failed to be important predictors of poor family functioning. Rather, previous family problems and low satisfaction with social support were the most broad and powerful predictors of poor functioning. To enhance the capabilities of families to manage the added responsibility of home monitoring, clinicians might wisely assess both of these aspects of family life and then target extra support services to families identified to be at risk in these areas. PMID- 8410390 TI - Intestinal infarction associated with milk curd obstruction in a preterm neonate. PMID- 8410391 TI - Bleeding as presenting symptom of cholestasis. AB - Four infants with conjugated hyperbilirubinemia who were brought for treatment primarily because of a hemorrhage are reported. Underlying disorders included extrahepatic biliary atresia, choledochal cysts, and alpha 1-antitrypsin deficiency. Prodromal signs of disturbed coagulation and diminished bile excretion were not recognized. The increased bleeding tendency was probably caused by vitamin K deficiency, resulting from a combination of cholestasis induced fat malabsorption, absence of vitamin K supplementation after birth, and low vitamin K intake as a result of breast feeding. PMID- 8410392 TI - Special imaging casebook. Neonatal adrenal hemorrhage and renal vein thrombosis. PMID- 8410393 TI - Ventilatory management casebook. Anterior pneumothorax-hyperlucent hemithorax sign. PMID- 8410394 TI - Predicting survival from in-hospital CPR: meta-analysis and validation of a prediction model. AB - OBJECTIVE: To better clarify patient factors that predict survival from in hospital cardiopulmonary resuscitation (CPR), using two methods: 1) meta-analysis and 2) validation of a prediction model, the pre-arrest morbidity (PAM) index. DESIGN: Meta-analysis of previously published studies by standard techniques. Retrospective chart review of validation sample. SETTING: University-affiliated teaching hospital. PATIENTS/PARTICIPANTS: Meta-analytic sample of 21 previous studies from 1965-1989. The validation sample consisted of all patients surviving resuscitation from the authors' hospital during the period September 1986 to January 1991. A matched sample of patients who did not survive from the same time period was used as the comparison group. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The strongest negative predictors of survival, by meta-analysis, were renal failure (r = 0.088, p < 0.0002), cancer (r = 0.08, p < 0.0002), and age more than 60 years (r = 0.063, p < 0.006). Sepsis (r = 0.046, p < 0.02), recent cerebrovascular accident (CVA) (r = 0.038, p < 0.04), and congestive heart failure (CHF) class III/IV (r = 0.036, p < 0.05) were weaker negative predictors. Presence of acute myocardial infarction (AMI) was a significant positive predictor of survival (r = 0.15, p < 0.0001). The PAM score was highly predictive of survival in a logistic regression model (p < 0.0003, R2 = 9.6%). No patient who survived to discharge had a PAM score higher than 8. CONCLUSION: Meta analysis reveals that the most significant negative predictors of survival from CPR are renal failure, cancer, and age more than 60 years, while AMI is a significant positive predictor. The PAM index is a useful method of stratifying probability of survival from CPR, especially for those patients with high PAM scores, who have essentially no chance of survival. PMID- 8410395 TI - Respiratory rate predicts cardiopulmonary arrest for internal medicine inpatients. AB - OBJECTIVE: To assess whether vital sign measurements could identify internal medicine patients at risk for cardiopulmonary arrest. DESIGN: Retrospective case control study comparing 72 hours of pre-arrest vital sign measurements with 72 hours of vital sign measurements for patients from the same units who did not experience cardiopulmonary arrest. SETTING: Twelve non-intensive care internal medicine units at a large midwestern academic medical center. PATIENTS: Cases included all 59 inpatients who had experienced cardiopulmonary arrest between May 1989 and December 1990; patients who were designated do-not-resuscitate (DNR) or had less than 72 hours of vital sign recordings were excluded. Controls included 91 inpatients without cardiopulmonary arrest who were matched for units and who had 72 hours of vital sign recordings. RESULTS: The occurrence of one or more respiratory rates > 27 breaths per minute over a 72-hour period had a sensitivity of 0.54 and a specificity of 0.83 (odds ratio = 5.56, 95% CL = 2.67-11.49) in predicting cardiopulmonary arrest. Other respiratory rate thresholds were also predictive of arrest. The ability of respiratory rate to predict arrest was stronger in units with high incidences of arrest relative to units with low incidences, for example, in units for the management of gastrointestinal disease (sensitivity = 1.00, specificity = 0.86) and renal disease (sensitivity = 0.69, specificity = 0.87). Respiratory rate remained a significant predictor (p < 0.001) after controlling for patient age and gender. Pulse rate and blood pressure were not predictive of cardiopulmonary arrest. CONCLUSIONS: Using elevated respiratory rates as a signal for focused diagnostic studies and therapeutic interventions in internal medicine patients may be useful in reducing the incidence of subsequent cardiopulmonary arrest, and lowering associated morbidity and mortality. PMID- 8410396 TI - Life-sustaining treatments during terminal illness: who wants what? AB - OBJECTIVE: To determine patient characteristics associated with the desire for life-sustaining treatments in the event of terminal illness. DESIGN: In-person survey from October 1986 to June 1988. SETTING: 13 internal medicine and family practices in North Carolina. PATIENTS: 2,536 patients (46% of those eligible) aged 65 years and older who were continuing care patients of participating practices, enrolled in Medicare. The patients were slightly older than the 65+ general population, 61% female, and 69% white, and most had one or more chronic illnesses. MEASUREMENTS AND MAIN RESULTS: The authors asked the patients whether they would want each of six different treatments (hospitalization, intensive care, cardiopulmonary resuscitation, surgery, artificial ventilation, or tube feeding) if they were to have a terminal illness. The authors combined responses into three categories ranging from the desire for more treatment to the desire for less treatment. After adjustment for other factors, 53% of women chose less treatment compared with 43% of men; 35% of blacks vs 15% of whites and 23% of the less well educated vs 15% of the better educated expressed the desire for more treatment. High depression scores also were associated with the desire for more treatment (26% for depressed vs 18% for others). CONCLUSION: Patients' choices for care in the event of terminal illness relate to an intricate set of demographic, educational, and cultural factors. These results should not be used as a shortcut to determine patient preferences for care, but may provide new insights into the basis for patients' preferences. In discussing choices for future life-sustaining care, physicians need to explore with each individual the basis for his or her choices. PMID- 8410397 TI - Variation in physicians' decision-making thresholds in management of a sexually transmitted disease. AB - OBJECTIVE: To gain insight into the variation in physicians' clinical decisions and further understand the factors that influence physicians' thresholds for testing and treating. DESIGN: Written clinical scenarios were mailed to two groups of physicians who were asked to provide probability estimates of syphilis, how these estimates might change with new information, and when a diagnostic test would be ordered or treatment begun. A model was then used to calculate the probabilities at which physicians ordered tests or initiated treatment. PARTICIPANTS: Group 1 comprised 126 board-certified internists from metropolitan Philadelphia responding from a sample of 360 such physicians randomly selected from a directory. Group 2 consisted of 31 experts in sexually transmitted disease responding from a sample of 50 experts selected by the authors. MEASUREMENTS AND MAIN RESULTS: Experts were willing to obtain a serologic screening test at a lower likelihood of syphilis (0.013%) than were internists (0.034%), and they were willing to obtain a lumbar puncture at a lower likelihood of neurosyphilis (0.165%) than were internists (0.393%). The difference in the groups' thresholds to begin neurosyphilis treatment was not significant. A multivariate model showed that group differences were created by individual characteristics (years in practice, subspecialty board certification, and full-time nonacademic practice) that were associated with higher thresholds for serologic screening. CONCLUSIONS: There are differences in the diagnostic testing practices for syphilis between national experts and internists. Although status in one of these groups alone did not predict the threshold for obtaining syphilis tests, certain individual characteristics were predictive. Examination of physician characteristics helps to explain the variation observed in their practice patterns, and determination of physicians' thresholds aids in analyzing these variations. PMID- 8410398 TI - Patient preferences: survival vs quality-of-life considerations. AB - OBJECTIVE: To assess whether patients can weigh risk comparisons involving mortality and quality of life in an understandable manner based on their willingness to accept risks of complications. DESIGN: Cross-sectional survey of patients. SETTING: University-based Department of Veterans Affairs Medical Center. PARTICIPANTS: 230 men patients seen in a general medicine clinic. MEASUREMENTS: Two survival graphs were used. Each graph contained survival curves for two alternative unidentified treatments for an unidentified medical condition. Graph 2 contained one curve that had a life expectancy that was 14% higher than the life expectancy of the corresponding curve in graph 1. Respondents were randomly assigned one of the two graphs and were asked to indicate which treatment they preferred and what risk of a change in their quality of life (urinary incontinence or importance) they were willing to accept to achieve longer survival. Patients were also asked whether they had a history of urinary incontinence or impotence. RESULTS AND CONCLUSIONS: Patients tended to be unwilling to accept worse quality of life to achieve increased survival over time. For both curve comparisons, significantly more (p < 0.01) patients accepted a treatment associated with higher mortality to avoid a 100% chance of incontinence than to avoid a 100% chance of impotence. Of the 75% (172/230) of patients reporting willingness to accept risk of either urinary incontinence or total impotence or both, 62% reported having at least some symptoms related to urinary incontinence or impotence. Of the 58 patients not willing to accept the complication risks, only 11% reported a history of urinary incontinence or impotence. The results show that patients are able to make distinctions about severity of morbidity, men are less willing to accept the risk of urinary incontinence than that of total impotence, and men patients who are symptomatic with urinary incontinence or impotence are more willing to accept the risks of treatment than are asymptomatic patients. PMID- 8410399 TI - A role for the primary care physician in counseling young African-American men about homicide prevention. AB - Homicide is the leading cause of death in African-American men aged 15-34 years, yet physicians rarely discuss homicide prevention with patients. The authors propose that physicians have a role in preventing homicide similar to their role in other preventive medicine issues. This study evaluated patients' responsiveness to a physician's counseling about firearms and homicide. While being treated for unrelated problems at a walk-in ambulatory clinic, 53 African American men patients received brief counseling by the physician about six preventive medicine topics, including firearms. A postvisit interview demonstrated that the discussion of firearms was well received and recalled more than any other preventive medicine issue discussed. PMID- 8410400 TI - Discriminating causes of dyspnea through clinical examination. PMID- 8410401 TI - Patient education through teaching for conceptual change. AB - It is not enough for clinicians to gather good patient information and then dictate management plans. If patient education is to be successful, attention must be paid to tailoring educational input to the patient's particular needs. If the conceptual change approach is followed, patient differences due to factors such as age and culture will be taken into account. Likewise, the different types of patient education described earlier can be accommodated, since the patient and his or her particular needs are always the focus of all medical conversations. The conceptual change approach for patient education potentially can help clinicians avoid the temptation to ignore patients' perspectives and provide instruction tailored to patient needs, thereby reducing the possibility of educational negligence. PMID- 8410402 TI - Life-prolonging treatments late in life. PMID- 8410403 TI - "Texas-sized" business cards for visually impaired patients. PMID- 8410404 TI - Subspecialty choice and faculty income. PMID- 8410405 TI - Should adult tetanus immunization be given as a single vaccination at age 65? A cost-effectiveness analysis. AB - OBJECTIVE: To compare three vaccination strategies for the prevention of adult tetanus. Each strategy includes childhood primary immunization and wound prophylaxis, and one of the following: 1) the currently recommended booster every ten years; 2) a single booster at 65 years of age; or 3) no intervention after age 6 except for wound prophylaxis. METHODS: Cost-effectiveness analysis was used to compare the three different strategies. A Markov model, cycled annually from age 5 through age 85, was applied to each strategy to predict the incidence and costs of tetanus for the U.S. adult population. RESULTS: The three strategies have very similar effects on life expectancy but different costs. Expressed incremental to no intervention after childhood primary immunization, the decennial booster strategy is least cost-effective, with a discounted incremental cost-effectiveness ratio of $143,138 per year of life saved compared with $4,527 for the single-booster strategy. Sensitivity analysis demonstrates that the decennial strategy is more effective but more costly over a wide range of model assumptions. CONCLUSIONS: The current policy of recommending tetanus booster vaccinations every ten years is effective but much more costly than a more easily implemented policy that also provides considerable protection against tetanus. The authors recommend forsaking decennial boosters in favor of a policy of including a single booster at age 65 along with other recommended health maintenance maneuvers reserved for that age. PMID- 8410406 TI - Is there a valid association between skin tags and colonic polyps: insights from a quantitative and methodologic analysis of the literature. AB - OBJECTIVE: To assess the reported association between skin tags and colonic polyps and to evaluate the methodologic rigor of the studies. DESIGN AND DATA IDENTIFICATION: English-language literature search using MEDLINE, Index Medicus, and bibliographic reviews of texts and all pertinent articles to perform a quantitative and methodologic analysis of all studies from 1983 (the original publication) assessing the association under study. Eligible studies were independently assessed using explicit methodologic guidelines for validity and generalizability of observational research. Two appraisers independently performed tests for heterogeneity and used meta-analytic methods in an attempt to provide summary estimates of the overall strength of association. MEASUREMENTS AND MAIN RESULTS: Significant statistical heterogeneity across studies indicated sharp differences in the direction and magnitude of the odds ratios for the association between skin tags and colonic polyps (chi-square test of homogeneity = 37.42, 9 degrees of freedom; p < 0.005). This marked disparity prevented meaningful pooling of the individual data. The degree of statistical heterogeneity was not significantly reduced after an analysis of specific subgroups of studies. Limitations potentially responsible for the varying outcomes included lack of blinded ascertainment of clinical information, noncomparability of subjects, differing diagnostic evaluations of the colon, and uncontrolled confounding. In addition, all but one study were performed in a tertiary care setting, seriously limiting the results to the "average" subject seen in primary care settings. CONCLUSION: Methodologic limitations and inconsistencies in study outcomes preclude the aggregation of data necessary to compute a valid and meaningful summary estimate of association. Sufficient variability prevents any consensus regarding the association between skin tags and colonic polyps. In addition, the applicability of the results is limited primarily to subjects seen in tertiary care centers, limiting the overall clinical usefulness of skin tags as "biomarkers" of colonic polyps. Recommendations for further research are provided. PMID- 8410407 TI - The characteristics of peer reviewers who produce good-quality reviews. AB - OBJECTIVE: To determine the characteristics of good peer reviewers. DESIGN: Cross sectional analysis of data gathered during a randomized controlled trial. SETTING: The Journal of General Internal Medicine. PARTICIPANTS: 226 reviewers of 131 consecutively submitted manuscripts of original research. 201 (91%) completed the review and submitted a curriculum vitae. MEASUREMENTS AND MAIN RESULTS: The quality of each review was judged on a scale from 1 to 5 by an editor who was blinded to the identity of the reviewer. Reviewer characteristics were taken from the curricula vitae. 86 of the 201 reviewers (43%) produced good reviews (a grade of 4 or 5). Using logistic regression, the authors found that when a reviewer was less than 40 years old, from a top academic institution, well known to the editor choosing the reviewer, and blinded to the identity of the manuscript's authors, the probability that he or she would produce a good review was 87%, whereas a reviewer without any of these characteristics had a 7% probability of producing a good review. Other characteristics that were significant only on bivariate analysis included previous clinical research training, additional postgraduate degrees, and more time spent on the review. There was a negative but statistically nonsignificant association between academic rank and review quality: 37% of full professors, 39% of associate professors, and 51% of assistant professors or fellows produced good reviews (p = 0.11). CONCLUSIONS: Good peer reviewers for this journal tended to be young, from strong academic institutions, well known to the editors, and blinded to the identity of the manuscript's authors. PMID- 8410408 TI - Doctors as workers: work-hour regulations and interns' perceptions of responsibility, quality of care, and training. AB - OBJECTIVE: To study residents' perceptions of their responsibility for patients, the quality of patient care, and their learning experiences in light of new work hour regulations. DESIGN: Inductive analysis of in-depth, semistructured, recorded interviews with a cohort of interns in internal medicine in the last month of their first postgraduate year. Questions were grounded in an examination of issues related to going off duty and delegating tasks to colleagues. Transcripts were independently analyzed by an interdisciplinary team. SETTING: New York University/Bellevue Hospital Center's residency program in internal medicine (in New York City). PARTICIPANTS: A cohort of 21 of a possible 24 interns in medicine on rotation at Bellevue Hospital Center. RESULTS: The interviews revealed: 1) intense concern harbored by interns for their patients with resulting difficulty in maintaining realistic boundaries between work and personal lives; 2) an open-ended workday and competing considerations confronting interns when deciding to leave the hospital--including concerns about leaving patients at critical junctures in their care, confidence in the colleague to whom they were signing out, regard for the workload of this colleague, and uneasiness about the educational consequences; 3) deterrents to acknowledging and acting on one's limits in performing medical work; and 4) a recurrent conflict between delegating responsibility and retaining control over patient care. CONCLUSION: Values traditionally learned in training emphasize autonomy and individual accountability. They may conflict with the shared decision making and collective responsibility among peers necessitated by work-hour limitations and associated changes in program structure. PMID- 8410409 TI - Psychiatric disorders and medical care utilization among people in the general population who report fatigue. AB - OBJECTIVE: To study the prevalence of fatigue in the general population and its association with psychiatric disorders, somatization, and medical utilization. SETTING: The public-use data tape from the 1984 National Institute of Mental Health Epidemiologic Catchment Area Study. PARTICIPANTS: Household sample of 18,571 subjects. INTERVENTIONS: Structured psychiatric interviews were reviewed to study the prevalence of complaints of current and lifetime fatigue and their relationship to selected psychiatric disorders. RESULTS: Fatigue has high current (6.7%) and lifetime (24.4%) prevalences in the general population. Medically unexplained fatigue also has high current (6.0%) and lifetime (15.5%) prevalences. When compared with those reporting no current fatigue, subjects who reported current (one-month) fatigue were significantly more likely to have experienced current and lifetime episodes of major depression, dysthymic disorder, panic disorder, and somatization disorder. They also had significantly higher mean numbers of lifetime and current DSM-III psychiatric diagnoses, medically unexplained physical symptoms (not just fatigue-related symptoms), and visits to health care providers than did patients without current episodes of fatigue. CONCLUSIONS: The high prevalence of fatigue in the general population appears to be significantly associated with increased lifetime and current risk for affective, anxiety, and somatoform disorders, as well as increased utilization of medical services. These data suggest that assessment of both medical and psychological health may be essential for the proper care of patients with fatigue. PMID- 8410410 TI - Sexual abuse history and women's medical problems. PMID- 8410411 TI - Effect of a new nootropic agent, CGS 5649B, on cognition, function, and behavior in dementia. AB - To determine the efficacy of CGS 5649B, the authors conducted a series of randomized, double-masked, single-subject studies using pairs of treatment periods, each pair including four weeks of active drug administration and four weeks of placebo administration. Thirty-two moderately demented patients completed three treatment pairs, receiving doses of 90, 200, or 600 mg of CGS 5649B during the active periods. Two patients met criteria for individual response in the behavioral measures. In the group analysis none of the differences achieved statistical significance. The authors conclude that CGS 5649B had a favorable impact on behavior for two of 32 patients. PMID- 8410412 TI - Utility of pulmonary function testing in the management of chronic obstructive pulmonary disease. AB - This study sought to measure the effect of pulmonary function testing (PFT) data on the decisions made by generalist physicians in the management of chronic obstructive pulmonary disease (COPD). 148 physicians were randomly assigned to two groups, both of which were asked to manage two identical fictitious but representative cases of COPD, which included history, physical, x-ray, and laboratory results. The experimental group received PFT results in addition. No significant difference was noted between the two groups in management based on availability of PFT data. The optimum utility of PFT data in the management of COPD may be exaggerated and has yet to be determined. PMID- 8410414 TI - Routine tetanus immunizations for adults: once is enough. PMID- 8410413 TI - Postmenopausal hormone replacement: are two hormones better than one? PMID- 8410415 TI - Relevancy of the ABIM examination. PMID- 8410416 TI - Effect of soft-tissue and joint injection with triamcinolone acetonide. PMID- 8410417 TI - HIV-related emergencies: frequency, diagnoses, and outcome. AB - OBJECTIVE: To study the clinical epidemiology and outcome of HIV-related emergencies, and to identify clinical predictors of HIV-related emergency hospitalizations. DESIGN: Case series. SETTING: Emergency facility of a tertiary care teaching hospital. PATIENTS: 350 HIV/AIDS patients followed at the authors' center. MEASUREMENTS AND MAIN RESULTS: 69 of 356 patients made 92 emergency visits with a frequency of 8% per month and 20% per quarter in a three-month study period. Forty-three visits (47%) resulted in hospitalization and contributed to 70% of total AIDS hospitalizations in the period. The five most common acute diagnoses were pneumonia (n = 22; 24%), fever (n = 15; 16%), upper respiratory infection (n = 9; 10%), cellulitis (n = 6; 7%), and gastroenteritis (n = 6; 7%). Three diagnoses accounted for 70% of acute HIV hospitalizations: pneumonia (n = 19), fever (n = 4), and sepsis (n = 4). Analysis of patient disposition as it relates to the patient's clinical presentation and HIV history using multivariate analysis yielded 1) the presence of dyspnea or cough (p = 0.015) and 2) fever with an abnormal chest x-ray (p = 0.008) as independently predictive of hospitalization. CONCLUSION: The findings indicate that HIV/AIDS patients have a frequent need for emergency care and most HIV/AIDS hospitalizations are emergency-related. The acute problems of these patients are related to a limited number of diagnostic categories, and the presence of respiratory or constitutional symptoms with an abnormal chest radiograph are the only reliable factors predictive of hospitalization. PMID- 8410418 TI - Dental disease among alcoholic individuals: a comparative study of hospitalized patients. AB - OBJECTIVE: To measure the presence and severity of dental disease, as assessed by physicians, among consecutively hospitalized alcoholic and nonalcoholic medical patients. DESIGN: Descriptive cross-sectional study. Patients who had Michigan Alcoholism Screening Test (MAST) scores > or = 8 were identified as being alcoholic. Nonalcoholic patients were defined as individuals with MAST scores < or = 4. These patients were matched with alcoholic patients for gender and age (+/- 5 years). The decayed, missing, and filled teeth (DMF) score and its components were used as a measure of dental disease. SETTING: General wards of internal medicine of a 1,000-bed urban teaching hospital in Lausanne, Switzerland. RESULTS: Among patients aged 20-75 years, the mean DMF score was higher among alcoholic patients than among nonalcoholic patients (26 vs 23, respectively; p < 0.001). This difference was greater among patients aged 20-39 years (20 vs 14, respectively) than among those aged 60-75 years (29 vs 27, respectively). The positive association between alcoholism and dental disease (crude odds ratio, 2.24; 95% CI, 1.15-4.31) remained after sequential stratification for several confounding factors. CONCLUSION: The study suggests that dental disease is frequent and severe in hospitalized medical patients and that alcoholism is an independent predictor of its severity. Routine assessment of dental disease by the physician is important for medical inpatients, especially among those who are alcoholic. PMID- 8410419 TI - Management and prevention of thromboembolic events in patients with cancer related hypercoagulable states: a risky business. AB - OBJECTIVE: To determine the optimal strategy for managing and preventing thromboembolic events in malignancy-associated hypercoagulable states. DESIGN: A Markov-based decision and cost-effectiveness analysis was performed. The authors explicitly considered consequences of embolic and bleeding events, filter complications, and cancer-related excess mortality. Data were drawn from the current literature. The main outcome measure for each strategy was the quality adjusted life expectancy and the total average variable costs. SUBJECTS: Patients with advanced malignancies prone to develop thromboembolic events, patients with acute proximal deep venous thrombosis (DVT), and patients who have survived a first episode of pulmonary embolism (PE). INTERVENTIONS: The authors considered three different interventions: 1) OBSERVATION, in which neither anticoagulant therapy nor filter placement is pursued, 2) ANTICOAGULATION, in which long-term anticoagulant therapy is started immediately, and 3) VENA CAVAL FILTER. MAIN RESULTS: Vena caval filter was the preferred strategy for every malignancy studied, yielding an 11% gain in quality-adjusted life expectancy, compared with observation, for patients with acute DVT, and an 18% gain for patients having survived a PE. Anticoagulant therapy yielded gains of 9% and 16%, respectively. Compared with anticoagulant therapy, filter was less costly due to the avoidance of additional expenses incurred by bleeding events. Prophylactic therapy was the least effective of the three strategies examined. CONCLUSIONS: Vena caval filter placement and long-term anticoagulation therapy yield similar outcomes in the setting of cancer-related hypercoagulable states. However, filter insertion is less expensive than anticoagulation. Given the short life expectancy and morbidity of patients with end-stage malignancy, patient preferences for health states must be considered in the decision-making process. If active treatment is pursued, vena caval filter should be used as a primary therapy. Prophylactic therapy is not warranted in any circumstance. PMID- 8410420 TI - Surgery for herniated lumbar discs: a literature synthesis. AB - OBJECTIVE: To assess the risks and benefits of surgery for herniated lumbar discs (discectomy) and to evaluate the methodologic quality of the literature. DESIGN: Literature synthesis. STUDY SELECTION AND DATA ANALYSIS: A structured MEDLINE search identified studies of standard, microsurgical, or percutaneous discectomy. Eligible studies had adult subjects, sample sizes of > or = 30, clinical outcome data for > or = 75% of patients, and follow-up of > or = 1 year. Summary rates of successful outcomes, reoperations, and complications were obtained by a random effects logistic regression model. Methodologic quality was assessed using established study design criteria. RESULTS: Eighty-one studies met inclusion criteria. Most had substantial design flaws and/or omitted important clinical data. Randomized trials of standard discectomy showed better short-term sciatica relief following surgery; 65% to 85% of patients reported no sciatica one year after surgery, compared with only 36% of conservatively treated patients. No data from randomized trials were available for microdiscectomy or percutaneous discectomy, although most outcomes appeared comparable to those of standard discectomy. Approximately 10% of discectomy patients underwent further back surgery, and rates increased over time. The rate of serious complications, including death and permanent neurologic damage, was less than 1%. CONCLUSIONS: Most studies were poorly designed and reported. Standard discectomy appears to offer better short-term outcomes than does conservative treatment, but long-term outcomes are similar. Discectomies are relatively safe procedures, though reoperations are common and increase over time. Decisions for elective surgery must balance faster pain relief against the risks and costs of surgery. PMID- 8410421 TI - A core component of the certification examination in internal medicine. AB - OBJECTIVE: To develop and test the psychometric characteristics of an examination of core content in internal medicine. DESIGN: A cross-sectional pilot test comparing the core examination with the 1988 certifying examination and two pretest examinations. SETTING: The 1988 certifying examination of the American Board of Internal Medicine. PARTICIPANTS: A random sample of 2,975 candidates from 8,968 candidates who took the 1988 certifying examination were given the core examination; similarly drawn samples were each given one of two pretests of traditional questions. INTERVENTIONS: A framework for developing an examination of core internal medicine questions was designed and used to develop a 92 question core test with an absolute pass/fail standard. RESULTS: Candidates answered 74% of core internal medicine questions, compared with 64%, 52%, and 53% of traditional questions on the 1988 certifying examination and the two pretests. The discriminating ability of the core internal medicine examination was lower than that of the certifying examination (r-values were 0.28 and 0.34, respectively). The pass rate was 83% for the core internal medicine examination and 57% for the certifying examination; 27% passed the core examination and failed the certifying examination; 1% passed the certifying examination and failed the core examination. CONCLUSION: Core internal medicine questions were easier than but almost as discriminating as traditional questions of the certifying examination. A small percentage of candidates passed the certifying examination but failed the core examination. PMID- 8410422 TI - Internal medicine housestaff and attending physician perceptions of the impact of the New York State Section 405 regulations on working conditions and supervision of residents in two training programs. AB - OBJECTIVES: To assess the attitudes of internal medicine housestaff and their attending physicians regarding the impact of the reduction in on-call working hours and increased supervision mandated in New York by a revision of the State Health Code (Section 405). DESIGN: Survey of senior medical housestaff and attendings two years after the adoption of the mandated changes. SETTING: Two independent medicine housestaff training programs of the Albert Einstein College of Medicine in the Bronx, New York. PARTICIPANTS: Fifty-three percent of third- and fourth-year residents (n = 79) and 60% of voluntary and full-time attendings (n = 266) responded. MEASUREMENTS: A factor analysis of 13 variables that appeared on both versions of the survey identified two interpretable factors. A multivariate analysis of variance compared responses to each factor by group and by campus, and Bonferroni post-hoc comparisons analyzed the items within factors. Chi-square analyses compared responses of residents and attendings to the open ended questions. RESULTS: Significant differences between the housestaff and attendings groups were found for all fixed-response items (minimum p < 0.05 for all analyses), but both groups agreed that the regulations had a positive impact on resident attitudes regarding the demands on their time. Both groups were also uncertain whether the new regulations had a beneficial effect on the choice of internal medicine as a career, the quality of resident supervision, and residents' intellectual interest in challenging medical problems. Whereas residents agreed that the regulations diminished their fatigue, had no impact on their ability to observe the full impact of interventions on patients, and resulted in better patient care, attendings were uncertain or disagreed. While attendings agreed that the regulations had caused a shift-work mentality among residents, housestaff were uncertain. CONCLUSIONS: Housestaff had more positive attitudes about the impact of the mandated changes in working conditions for residents than did attending physicians in the same institutions. The major benefits seen by residents were less fatigue and more spare time. There was no consensus about whether these changes had a positive impact on internal medicine practice and clinical supervision. There was some concern that a shift-work mentality is developing among residents and that continuity of patient care has suffered. Thus, despite some substantial benefits, Section 405 may not be achieving its goals of improving resident supervision and the quality of patient care by houseofficers. PMID- 8410423 TI - The tyranny of the throwaways. PMID- 8410424 TI - Reserpine reconsidered: no association with serious peptic ulcer disease. AB - A history of peptic ulcer disease is frequently cited as a contraindication to the use of reserpine. However, the risk of ulcer disease associated with the use of reserpine at current therapeutic doses is unknown. To address this question, the authors conducted a nested case-control study of the association between reserpine use and hospitalizations for peptic ulcer disease in elderly Tennessee Medicaid enrollees. When compared with that of nonusers of reserpine, the relative risk of hospitalization for peptic ulcer disease was 0.8 (95% CI, 0.6 1.0) among current users and 0.8 (95% CI, 0.5-1.3) among former users. The authors' data provide evidence that reserpine is not associated with ulcer disease in elderly persons and suggest that a history of ulcer disease need not be a contraindication to the use of this drug. PMID- 8410425 TI - Educational characteristics of ambulatory morning report. AB - The educational characteristics of ambulatory morning report were compared with those of the inpatient morning report sessions over a five-month period. Ambulatory morning report had fewer total participants and was more likely to cover general internal medicine topics (p < 0.05), the medical interview (6.8% vs 1.2%, p = 0.02), and social issues (9.6% vs 1.2%, p = 0.02). Morning report conference can be replicated in the ambulatory setting, thus providing an opportunity to discuss general medicine topics not usually addressed in the inpatient setting. PMID- 8410426 TI - Resident recognition of open-angle glaucoma: effects of an educational intervention. AB - To test an educational intervention's effect on improving detection of glaucoma by direct ophthalmoscopy, 14 medicine residents examined five patients, two with ophthalmoscopic changes of glaucoma and three with normal fundi. The residents observed a standardized slide/narrative educational intervention reviewing glaucomatous ophthalmoscopic changes and then re-examined the same patients eight to 12 weeks later. The intervention's odds of improving residents' diagnostic impression were significant (OR = 2.2; 95% CI = 1.3-3.6), with significant improvement in sensitivity (p = 0.02) and a trend toward improved specificity. These findings confirm that the diagnosis of glaucomatous ocular changes on eye examinations by medicine residents can be improved with a brief educational intervention. PMID- 8410427 TI - Testosterone for erectile failure. PMID- 8410428 TI - Exercise guidelines. PMID- 8410429 TI - Violence: an epidemic that's right at home. PMID- 8410430 TI - Nursing management of umbilical cord prolapse. AB - Advances in the management of umbilical cord prolapse hold promise for reducing perinatal morbidity and mortality. This article reviews traditional nursing management of umbilical cord prolapse and introduces the use of real-time ultrasound and bladder filling as additional methods of nursing management. A case report is provided. PMID- 8410431 TI - Prenatal and postpartum care in Hawaii: a community-based approach. AB - Given the problems of access, retention, and relevant prenatal care content, supplements to existing programs for health-care delivery during pregnancy and after birth are necessary. This article describes a community-based approach to prenatal and postpartum care that has been developed to address these issues. Culturally sensitive strategies were created for use with Hawaiian, Filipino, and Japanese women living on the island of Hawaii. Six nursing care and community outreach interventions were used. Local public health nurses assisted in developing the program and are responsible for its coordination and implementation. PMID- 8410432 TI - Adherence to ACOG guidelines on exercise during pregnancy: effect on pregnancy outcome. AB - OBJECTIVE: To determine the extent to which exercising women adhered to the ACOG guidelines on exercise during pregnancy and the effect of nonadherence on pregnancy outcome. DESIGN: Nonexperimental, retrospective survey. SETTING: Two national physical fitness conferences. PARTICIPANTS: One hundred seventy-three women who exercised during their pregnancies. OUTCOME MEASURES: Maternal weight gain, neonatal birth weight, occurrence of cesarean delivery, and gestational age of newborn. RESULTS: Eighty-three percent of the women knew of the ACOG guidelines. A total of 87 (53%) adhered to the guidelines. Women who exercised with heart rate intensities above 150 beats per minute, for durations longer than 40 minutes through the 3rd trimester, or both had no significant difference in maternal weight gain, fetal birth weight, or gestational age of the newborn (p > .05) than those who exercised more moderately. The long-duration and moderate intensity groups had significantly fewer cesarean deliveries (p < .05). CONCLUSION: Women who exercised regularly before conceiving and who had uncomplicated pregnancies did not adversely affect their own or their offsprings' health by exercising in excess of the ACOG guidelines. PMID- 8410433 TI - Breastfeeding support services in the neonatal intensive-care unit. AB - OBJECTIVE: To describe a model for providing breastfeeding support in the neonatal intensive-care unit (NICU). DESIGN: Naturalistic, participant observation. SETTING: Suburban Level III NICU. PATIENTS: One hundred thirty-two mother-infant pairs over 1 year. Infants were hospitalized in the NICU, and mothers had initiated lactation efforts. INTERVENTIONS: Investigators provided breastfeeding interventions for the mother-infant pairs, based on identified problems, the research literature, or both. MAIN OUTCOME MEASURES: Percentage of mothers who were breastfeeding at the time of discharge from the NICU. RESULT: Interventions were classified into five categories: expression and collection of mothers' milk, gavage feeding of expressed mothers' milk, in-hospital breastfeeding sessions, postdischarge breastfeeding management, and additional consultation. CONCLUSIONS: This model was effective in preventing breastfeeding failure for this population. The model can provide the basis for NICU breastfeeding standards of care, protocols, and chart records, or for reimbursement purposes. The model also provides a framework for studying a specific category or breastfeeding intervention. PMID- 8410434 TI - Intravenous meperidine infusion for obstetric analgesia. AB - OBJECTIVE: To provide safe, satisfying obstetric analgesia when epidural analgesia was unavailable. DESIGN: Randomized, controlled study of women in labor. SETTING: A regional hospital offering primary- and secondary-level obstetric care. PARTICIPANTS: Thirty-nine women with low-risk, single, term pregnancies and anticipated vaginal delivery. INTERVENTIONS: Women in the intermittent intramuscular (IM) injection group received meperidine 50-100 mg, every 2 hours, as required. Those in the intravenous (IV) infusion group received an initial bolus of meperidine 25 mg, a basal infusion rate of 60 mg/h, and intermittent boluses of 25 mg/h as required. Each participant had a maximum of 200 mg of meperidine available to her during labor. MAIN OUTCOME MEASURE: Pain intensity was measured on a 10-cm visual analogue scale. Pain ratings were recorded when analgesia was initiated and every 30 minutes thereafter, until delivery. RESULTS: Women who received IV meperidine reported significantly lower levels of pain (p = .0015) than women in the IM group. However, women in the IV group received significantly higher doses of meperidine (mean = 121 mg) than those in the IM group (mean = 82 mg; p = .0007). When pain scores were compared on a smaller group of participants (n = 18) who received similar doses of meperidine (100-150 mg), women in the IV group still reported lower pain scores (p = .0092). No significant differences were found between the groups in length of labor, maternal vital signs, fetal heart rate, Apgar scores, level of maternal sedation, or patient-reported side effects. CONCLUSION: IV meperidine infusion was superior to intermittent IM injections for pain relief during labor. PMID- 8410435 TI - Fathers of breastfed infants: postponing and types of involvement. AB - OBJECTIVE: To examine husbands' experiences of having their wives breastfeed. DESIGN: Qualitative method of grounded theory. SETTING: In-depth interviews by phone at home. PARTICIPANTS: Fourteen middle-class, urban, Canadian fathers of successfully breastfed infants. RESULTS: The fathers reported a disparity in the types of relationships that their children had with each of their parents as a result of breastfeeding. The process that enabled the fathers to accept this perceived difference was labeled postponing. Four fathering styles, which explain the variation in postponing, were found. The phases of postponing include becoming aware of the disparity, simultaneously developing accepting strategies and acknowledging reinforcing factors, and, finally, developing compensating behaviors to increase the fathers' interactions with their infants and promote closer relationships. When weaning occurred, the fathers' relationships with their children changed as the fathers found ways to catch up. CONCLUSIONS: A need exists for realistic education about the realities of breastfeeding for fathers. More outlets for fathers' negative emotions toward breastfeeding need to be found. PMID- 8410436 TI - The relationship between women's symptoms of endometriosis and self-esteem. AB - OBJECTIVE: To determine the relationship between women's symptoms of endometriosis and self-esteem. DESIGN: Descriptive. SETTING: Private practice. PARTICIPANTS: Twenty-three mostly middle class, employed women who had been diagnosed with endometriosis and had received treatment from a physician. METHOD: The volunteer subjects completed a two-part questionnaire. One part consisted of questions on demographic characteristics, health history, menstrual history, and symptoms of endometriosis, and the other part consisted of the Rosenberg Self Esteem Scale. MAIN OUTCOME MEASURES: The relationship between women's self-esteem and perceptions of the number, frequency, and severity of their symptoms. RESULTS: No significant correlation was found between self-esteem and the number, frequency, and severity of endometriosis symptoms. Significant correlations were found between the number/frequency of symptoms and the severity of pain and between the frequency and number of symptoms. CONCLUSION: Although endometriosis symptoms did not influence self-esteem, they interfered with women's lives. Assessment of psychosocial status and nursing interventions designed to relieve pain are important factors in assisting women with endometriosis. PMID- 8410437 TI - Surface refractive index of the eye lens determined with an optic fiber sensor. AB - The use of a fiber-optic sensor for measurement of the refractive index on the surface of eye lenses is described. The technique makes use of the fact that the amount of light reflected at the interface of two media (Fresnel reflectance) depends on the refractive-index difference between them. The sample is probed with a single-mode fiber, and the refractive index is calculated from the proportion of light reflected at the probe-sample interface. PMID- 8410438 TI - Eccentricity and the Ferry-Porter law. AB - Is the flicker limit governed by the Ferry-Porter law or by some other law? We previously obtained strong evidence for the wide applicability of the Ferry Porter law and for its variation with eccentricity [J. Opt. Soc. Am. A 7, 743 (1990)]. Raninen and others [Vision Res. 28, 785 (1988); 31, 1875 1875 (1991)] have questioned our analysis of the data and our conclusions. We show that their criticism of the Ferry-Porter law is based on specious analyses and that their own data support the Ferry-Porter formulation. We also provide further evidence that the slope of the Ferry-Porter function increases markedly with eccentricity, implying that the inherent temporal properties of the retina grow more rapid with distance from the fovea. PMID- 8410439 TI - Rx for IUSD: get lean and mean. Faculty reorganizes school structure and curriculum. Interview by Robert P. Carlson. PMID- 8410440 TI - Utilization of 3-D computer tomographs in oral surgery. AB - Imaging is essential in the diagnosis and treatment of maxillofacial abnormalities. CT scans are but one type of imaging studies available to the practitioner. Computers have recently enhanced the traditional CT scan so that it can now be viewed in a three-dimensional reconstruction. Essentials of 3-D imaging and its uses will be presented by the authors, with various case reports to illustrate their value in the dental office. PMID- 8410441 TI - Guided tissue regeneration: the new frontier in periodontics. AB - The ideal goal of periodontal therapy is the reestablishment and regeneration of the lost periodontal tissues. Many different treatment regimens have been suggested to achieve this dream; however, most are unpredictable and highly debated in the periodontal literature. This paper discusses the most current philosophy in periodontal regeneration--guided tissue regeneration. A brief review of the literature, surgical technique, and case presentations pertaining to this procedure are provided. PMID- 8410442 TI - Considering the patient as well as the problem. PMID- 8410443 TI - Management of ameloblastoma: a changing perspective. PMID- 8410444 TI - An alternate elective neck incision. AB - The classic submandibular incision parallels the inferior border of the mandible and does not follow the resting skin tension lines of the neck. A modified approach is described that initially follows these lines but, as the midbody region is approached, a zigzag incision with legs of 1 to 3 cm and tip angles of approximately 70 to 90 degrees is made. This zigzag skin incision is adjusted for the intended surgery. Subjectively, the results are much more esthetic. PMID- 8410445 TI - A comparison of propofol with methohexital and isoflurane in two general anesthetic techniques. AB - The purpose of this study was to compare two general anesthetic techniques involving oral intubation for use in outpatient third molar surgery. Fifty American Society of Anesthesiologists I or II patients were randomly allocated to two groups. Group 1 received methohexital, isoflurane, nitrous oxide, and alfentanil, while group 2 received propofol, alfentanil, and nitrous oxide. An analysis of the results showed that although the technique used in group 2 cost more, had a slightly longer induction time, and produced a similar duration of apnea, it did not cause significant hypotension (as previously reported), and had a significantly better overall recovery. It was concluded that the use of propofol in the outpatient surgery setting may be a valuable addition to the oral and maxillofacial surgeon's anesthetic armamentarium. PMID- 8410446 TI - The use of ultrasound to determine the position of the mandibular condyle. AB - A randomized, single blind study of 20 patients examined the accuracy of ultrasound in establishing the position of the mandibular condyle in relation to the glenoid fossa. The sonographic technique is described. The temporomandibular joint was imaged sonographically with the patients in an open- and a closed-mouth position as a model for condylar sag and proper condylar seating, respectively, during mandibular ramus osteotomy procedures. One radiologist identified condylar position correctly in 38 of 40 still ultrasound images, with a sensitivity and a specificity of 95%. During real time ultrasound examination, it is possible to visualize varying degrees of condylar movement in relation to the glenoid fossa. The results of this study support the potential use of ultrasound as an adjunct to mandibular orthognathic surgery. PMID- 8410447 TI - Histologic study of articular cartilage repair in the marmoset condyle. AB - This study tests the regenerative capability of condylar cartilage by examining repair of full-thickness articular surface defects in a primate model. The wounds were reconstituted within 6 months with tissue identical to the remaining condylar structure, and the reformed articular surface was maintained intact up to 1 year postinjury. PMID- 8410448 TI - Bone regeneration by the osteopromotion technique using bioabsorbable membranes: an experimental study in rats. AB - An osteopromotive technique has been developed that allows improved bone regeneration as well as bone neogenesis using porous, inert, nondegradable membranes made of expanded polytetrafluoroethylene (e-PTFE). For certain applications, however, it would be advantageous to use bioabsorbable membranes (BAMs), thus avoiding a surgical reentry for membrane removal. In this randomized comparison study the osteopromotive potential of BAMs was investigated in standardized "critical size defects" (5 mm in diameter) in the rat mandible. Three membrane types were tested and comparisons were made with e-PTFE membrane. The BAMs consisted of polylactic/polyglycolic acid copolymers designed to give the membranes different absorption times when implanted in the tissue. Histologic analysis after healing periods of 1 to 12 weeks demonstrated the BAMs to be well tolerated by the tissue, causing just a mild inflammatory reaction along the membrane surfaces as long as the material remained in the tissue. The BAMs were found to be as efficient as e-PTFE membranes in that the bone repair was not significantly different with any of the four membrane types. However, healing in conjunction with one type of BAM seemed to occur somewhat more rapidly. Some cartilage was present at the early healing stages in the defects treated with BAMs, but disappeared at later stages. The results of this study show that BAMs are a valid alternative to e-PTFE membranes to improve bone regeneration, but indicate that further technical development of the membrane material is necessary. PMID- 8410449 TI - Experimental temporomandibular joint disc perforation in the rabbit: a gross morphologic, biochemical, and ultrastructural analysis. AB - This study evaluates the progression of osteoarthritis (OA) in the adult New Zealand white rabbit temporomandibular joint following unilateral disc perforation. Thirty-seven animals were divided into five groups: control (n = 8), 6-week sham (n = 5), and experimental 6-, 12-, and 24-weeks (n = 8 each). Quantitative data was examined with two-way analysis of variance, and followed by Scheffe pair-wise comparisons. Transmission electron microscopy, acid phosphatase [AcP] activity, uronic acid content, and gross morphologic analysis indicated that disc perforation induced remodeling activity and degenerative changes in the condylar cartilage and bone as early as 6 weeks postoperatively. AcP activity of homogenized cartilage samples was significantly increased in experimental joints versus the side that did not undergo surgery at 6 and 12 weeks (P < .05). Uronic acid content was significantly greater in experimental joints versus the side that did not undergo surgery at 6 weeks (P < .05). Heightened cellular activity was present in the deep zone of osteoarthritic fibrocartilage of the 6- and 12 week experimental groups. Degenerating chondrocytes appeared to contain greater proportions of intracytoplasmic filaments and lysosome-like bodies. Disc perforation provided the impetus for degenerative or remodeling changes in the condylar cartilage of experimental joints, and is consistent with secondary OA. These dynamic events were most significant in the deep zone of articular fibrocartilage. PMID- 8410450 TI - Effects of photodynamic therapy in the normal mouse tongue. AB - The effects of photodynamic therapy (PDT) on the normal mouse tongue were investigated. After using various doses (2.5, 10, and 20 mg/kg body wt) of the photosensitizing drug hematoporphyrin oligomers and 90 or 180 J/cm2 red light (630-nm) emitted from a pulsed neodymium:yttrium-aluminum-garnet dye laser was used to activate it 3 or 24 hours later. It was found that the 20-mg/kg dose elicited a severe response that included extensive vesicular and edema formation. A less severe response was observed with 10 mg/kg of the drug and low-power light (5 mJ/cm2/pulse) periodically delivered (1 hour interval between two 30-minute photoradiations). Such a regimen, however, produced more damage when compared with the higher power (15 mJ/cm2), continuous light delivery counterpart. Healing, except for the protocol with only a 3-hour drug-light interval, was attained by 5 days post-PDT as indicated by regeneration with histologically normal tissues and quantitatively a return to untreated, control values for cross sectional areas and number of blood vessels. Bromodeoxyuridine immunohistochemistry disclosed an immediate increase in the labeling indices, ie, the percentage of S phase cycling cells, indicating stimulated cell proliferation secondary to repair and fast repopulation of the epithelium. Under the commonly used protocols, PDT was provided safely to the mouse tongue. The regimen of low drug dose and low power of light periodically delivered appears to be the most acceptable method. These parameter-dependent results may partly form the basis for the judicious application of PDT to the oral cavity. PMID- 8410451 TI - Manpower excess: one source of our problems. PMID- 8410452 TI - Soft tissue swelling in the mandibular buccal vestibule. PMID- 8410453 TI - Air emphysema secondary to mandibular fracture: case report. PMID- 8410454 TI - Accessory jaw bone: report of a case. PMID- 8410455 TI - Salvage of a microvascular scalp reconstruction with Hirudo medicinalis. PMID- 8410456 TI - Malignant (metastatic) ameloblastoma: report of a case. PMID- 8410457 TI - Microscrew fixation of zygomatic arch fractures. PMID- 8410458 TI - Protection of the airway against foreign body aspiration. PMID- 8410459 TI - Recommendations for management of patients with temporomandibular joint implants. Temporomandibular Joint Implant Surgery Workshop. PMID- 8410460 TI - Treatment of experimental osteomyelitis with antibiotic-impregnated bone graft substitute. AB - The model of Norden was used to induce osteomyelitis in the left tibia of New Zealand White rabbits. Twenty-one days following inoculation, the animals had primary debridement and then were randomized into one of three treatment groups. Group I received no additional treatment; in Group II, plain hydroxyapatite beads were packed into the defect; and in Group III, gentamicin crobefat-loaded hydroxyapatite beads were packed into the defect. The animals were observed for 40 days after the primary debridement and then were killed. The intensity of infection was determined by swab cultures and quantitative bacterial cultures of the debrided material. At primary debridement, all of the animals in each group were equally infected. At the time of secondary debridement, only the animals in Group III had a statistically significant reduction in infection (p < 0.001). In this study, we demonstrated that an antibiotic-loaded osteoinductive ceramic bead can effectively eliminate bacteria from an osteomyelitic cavity. PMID- 8410461 TI - In vitro pharmacokinetics of antibiotic release from locally implantable materials. AB - Local deposition of antibiotics has became increasingly popular in the management of open fractures or osteomyelitis, and several substances have been employed as the vehicle for delivery. Although the elution characteristics of some substances have been documented, a comparative study of the characteristics of the commonly used substances could establish the clinical indications for particular vehicles. Cylindrical pellets of uniform size (6 x 4 mm) were prepared from bone graft (BG), demineralized bone matrix (DBM), plaster of Paris (POP), or polymethylmethacrylate (PMMA), with 25 mg of tobramycin/g of substance in each pellet. The pellets were suspended in phosphate buffered saline, and the antibiotic concentration in the buffer was determined at various time intervals by an enzyme immunoassay. BG and DBM eluted 70 and 45% of their antibiotic load by 24 h, and negligible amounts were detected at 1 week; POP released 17% of its load by 24 h, with trace amounts detected at 3 weeks; and PMMA eluted 7% at 24 h, with trace amounts detected for as long as 14 days. These findings suggest that the optimal vehicle for local deposition of antibiotic depends on the clinical setting. BG and DBM may be best employed when brief antibiotic coverage is required (as for acute contaminated open fractures), whereas POP and PMMA may be better suited for long-term coverage (such as for established osteomyelitis). PMID- 8410462 TI - In vitro photodynamic therapy of musculoskeletal neoplasms. AB - Photodynamic therapy is a tumoricidal modality that utilizes an inactive pharmacologic agent that becomes activated on exposure to visible light. Neoplasms selectively retain and accumulate photosensitizers at levels generally higher than surrounding non-neoplastic tissues. The purpose of this study was to establish a testing method for in vitro investigation of the effects of photodynamic therapy on human musculoskeletal neoplasms by examination of the sensitivity of these tumors to photoactivation. Three human musculoskeletal neoplasms were cultured, exposed to the photosensitizer Photofrin, and then studied for their response to photodynamic therapy after laser activation. Giant cell tumor, dedifferentiated chondrosarcoma, and osteosarcoma were examined with use of strict experimental controls. The photoradiation conditions during photodynamic therapy were kept constant. Cell viability was determined as a function of energy dose. We concluded that the three musculoskeletal tumors were susceptible to in vitro photodynamic therapy and the test system was reproducible. The optimal in vitro nontoxic incubation concentration of Photofrin was 3 micrograms/ml. A differential cytotoxic response to photodynamic therapy was exhibited by the musculoskeletal neoplasms as a function of increased dosages of energy. PMID- 8410463 TI - Effects of ascorbic acid, calcitriol, and retinoic acid on the differentiation of preosteoblasts. AB - The responses of the immortalized rat preosteoblast UMR-201-10B to ascorbic acid (AA), 1,25(OH)2D3 (calcitriol), and retinoic acid (RA) were examined. UMR-201-10B cells have an undetectable basal alkaline phosphatase (ALP) activity that is induced after 24 h of treatment with 10(-6) M RA (4.64 +/- 0.06 mumol/h/mg of protein). The addition of 10(-8) M calcitriol resulted in a slight induction of ALP activity after 72 h (0.43 +/- 0.07 mumol/h/mg of protein). When calcitriol was added to RA, however, over the same period ALP activity was enhanced significantly compared with treatment with RA alone (RA and calcitriol, 12.29 +/- 0.86 mumol/h/mg of protein). Treatment with AA (50 micrograms/ml) alone had no effect on ALP activity but increased RA-induced ALP activity to 6.78 +/- 0.28 mumol/h/mg of protein at 24 h. In contrast, AA inhibited calcitriol-induced ALP activity after 7 days of combined treatment with calcitriol (calcitriol, 7.73 +/- 0.16 mumol/h/mg of protein; AA and calcitriol, 1.44 +/- 0.06 mumol/h/mg of protein). Individually, RA and calcitriol induced mRNA expression for ALP, matrix gla protein (MGP), and osteopontin (OP). The steady state level of pro-alpha 1(I) collagen mRNA also was increased significantly by treatment with RA and AA individually. The combination of RA and calcitriol had a synergistic effect on ALP, OP, and especially MGP mRNA expression but significantly reduced the expression of pro-alpha 1(I) collagen mRNA. AA enhanced the effect of RA on the expression of pro-alpha 1(I) collagen, MGP, and ALP mRNAs as well as the effect of calcitriol on OP and MGP. The addition of AA to RA resulted in a decrease in the steady state level of OP, whereas its cotreatment with calcitriol caused a decrease in pro-alpha 1(I) collagen and ALP mRNA. In conclusion, these studies identify RA, calcitriol, and AA as regulators of differentiated osteoblast function. PMID- 8410464 TI - Effect of ischemic storage on endothelium in canine bone. AB - The limits of room temperature (24 degrees C) and hypothermic (4 degrees C) storage for preservation of endothelial eccrine function were investigated with use of an ex vivo perfusion apparatus model of the cannulated canine tibia. Norepinephrine-induced contraction of smooth muscles in both the presence and absence of acetylcholine was compared, with and without selective blockade of endothelial-derived relaxing factor (EDRF). At room temperature, release of EDRF deteriorated after 24 h and was absent after 48 h of storage. At 4 degrees C, endothelial eccrine activity appeared unimpaired for as long as 72 h of storage. From 96-192 h of storage at 4 degrees C, release of EDRF was suggested, but with an attenuation of its effect on smooth muscle. At 216 and 504 h of storage at 4 degrees C, no endothelial eccrine activity was demonstrated. PMID- 8410465 TI - Enhancement of bone growth into metal screws implanted in the medullary canal of the femur in rats. AB - The kinetics of growth of bone into control (nontreated) and heat-treated screws made of stainless steel (type 316L) and Ti-6Al-4V, implanted in the medullary canal of the femur in rats, were studied by mechanical, histological, and biochemical methods. A progressive and significant increase in the ingrowth of bone, as reflected by interfacial shear strengths of the screws, was measured with time after implantation. At all time intervals for as long as 35 days after implantation, the shear strength of the heat-treated Ti-6Al-4V and stainless steel screws was significantly higher than (1.6-3.4 times) that of the control screws. The specific activity of alkaline phosphatase in extracts of tissue from around the implanted screws peaked 6 days after insertion, with significantly higher values at 5, 6, and 7 days postoperatively for the heat-treated screws than for the controls. The extent of calcification also was higher at all time intervals for the heat-treated screws than for the controls. The histological evaluation of formation of bone between the ridges of the implanted screw corroborated the mechanical and biochemical measurements. At each time interval, a more mature bone was noted around the heat-treated screws than around the controls. It was concluded that the heat treatment of metal implants before insertion can result in augmentation of osseous ingrowth 1.6-5.3 times that into control implants in an in vivo experimental model. PMID- 8410466 TI - Pulsed magnetic fields improve osteoblast activity during the repair of an experimental osseous defect. AB - The influence of pulsed low-frequency electromagnetic fields (PEMFs) on bone formation was investigated in studies of the healing process of transcortical holes, bored at the diaphyseal region of metacarpal bones of six adult horses, exposed for 30 days to PEMFs (28 G peak amplitude, 1.3 ms rise time, and 75 Hz repetition rate). A pair of Helmholtz coils, continuously powered by a pulse generator, was applied for 30 days to the left metacarpal bone, through which two holes, of equal diameter and depth, had been bored at the diaphyseal region. Two equal holes, bored at the same level in the right metacarpal and surrounded by an inactive pair of Helmholtz coils, were used as controls. All horses were given an intravenous injection of 25-30 mg/kg of tetracycline chloride on the 15th and again on the 25th day after the operation and were killed 5 days later. The histomorphometric analysis indicated that both the amount of bone formed during 30 days and the mineral apposition rate during 10 days (deduced from the interval between the two tetracycline labels) were significantly greater (p < 0.01 and p < 0.0001, respectively) in the PEMF-treated holes than in the controls. As did a previous investigation, these preliminary findings indicate that PEMFs at low frequency not only stimulate bone repair but also seem to improve the osteogenic phase of the healing process, at least in our experimental conditions. PMID- 8410467 TI - Associations and dissociations between serum bone Gla protein and alkaline phosphatase in skeletal metabolism. AB - Serum bone Gla protein (BGP) and alkaline phosphatase (AP) activity were compared for the assessment of skeletal status in 11 normal pregnant women, 12 normal women on days 3 and 13 of the menstrual cycle, five postmenopausal women before and after 1 month of treatment with ethinyl estradiol (20 micrograms/day), five patients with cancer and hypercalcemia during treatment with calcitonin, and one patient with Paget disease during treatment with Plicamycin. BGP and AP correlated with each other only in the pregnant women. In all other circumstances, there was no correlation between these two serum osteoblast products. Furthermore, there were conditions in which the two measurements became discordant. These studies demonstrate that BGP and AP commonly are dissociated when used as measurements of skeletal status. Although both are osteoblast products, BGP and AP probably reflect different aspects of osteoblast differentiation and function. PMID- 8410468 TI - Ultrastructural immunolocalization of type-VI collagen and chondroitin sulphate in ligament. AB - Immunological methods were used to determine the identity of the major components comprising a network of electron-dense seams (described by the authors in a previous work) within the extracellular matrix of medial collateral ligament (MCL) from humans and rabbits. Tissue obtained from MCL midsubstance was subjected to pre-embedding labelling with colloidal gold at the electron microscopic level with monoclonal antibodies (MAbs) against type-VI collagen and chondroitin sulphate (CS), before and after digestion with chondroitinase ABC and testicular hyaluronidase. Tissue labelled with anti-type-VI MAbs showed gold conjugates attached to the microfilamentous component of the seams both before and after enzyme digestion, which confirmed the identity of the beaded microfilaments as type-VI collagen. Treatment of the tissue with anti-CS MAbs resulted in labelling of undigested tissue only. In these treatments, gold particles were found attached to granules that were interspersed throughout the network of type-VI microfilaments. Both the granules and gold labels were absent from the network following enzyme digestion. Thin nonbeaded microfilaments that did not label with anti-type-VI MAbs also were present within the seams. The loss of these nonbeaded microfilaments following enzyme digestion suggested that they might represent strands of hyaluronan. The codistribution and sequestering of type-VI collagen and CS within discrete seams or channels suggests that these regions of the MCL midsubstance may contain higher concentrations of water than the surrounding dense fibrillar matrix. PMID- 8410469 TI - Determination of the in situ loads on the human anterior cruciate ligament. AB - A noncontact, kinematic method was used to determine the lengths and in situ loads borne by portions of the human anterior cruciate ligament (ACL) by the combination of kinematic data from the intact knee and load-length curves of the isolated ACL. Specimens from knees of cadavers of young people were tested in passive flexion and extension as well as with 100 N of anterior tibial drawer at 0, 30, 45, and 90 degrees of flexion. The results showed that the in situ load on the whole ACL (as much as 129 N) can exceed the magnitude of the applied anterior tibial drawer. The load distribution within the ligament changes with flexion of the knee. The anterior and posterior portions share the anterior drawer force equally toward full extension. However, at flexion > 45 degrees, the anterior portion supports 90-95% of the load. This information is important for the determination of the function of the entire ACL and of its subportions in response to external loading of the intact knee. In particular, the preferential loading found for one of the portions of the ACL demonstrates that successful operative reconstruction of this ligament may not be achieved simply by reproduction of its gross anatomy; consideration of the role of the ligament in the overall kinematics of the knee is necessary. PMID- 8410470 TI - Effect of reconstruction of the anterior cruciate ligament on proprioception of the knee and the heel strike transient. AB - Abnormal proprioception of the knee joint has been documented after rupture of the anterior cruciate ligament (ACL) and may result in the loss of muscular reflexes. Excessive loading from the lack of muscular control may predispose the joint to osteoarthrosis. To investigate this problem, 10 patients were studied at an average of 31.6 months after ACL reconstruction. Three tests of joint proprioception and measurements of the vertical component of heel strike force during normal gait were used. A normal control group also was studied. For two of the proprioception tests (reproduction of passive motion and relative reproduction), there were no statistical differences among the uninjured (control) limbs, the normal contralateral limb of patients with a reconstructed ACL, and the extremity with a reconstructed ACL. In the third test (threshold of detection of motion), which previously has been shown to be adversely affected by ACL injury, the measurements for both extremities of patients with a reconstructed ACL were more accurate than those for the control group. The reconstructed extremity performed less accurately than the contralateral extremity (p < 0.05). The heel strike transient (vertical component of ground reaction force at heel strike) for uninjured and ACL-reconstructed limbs was not significantly different. In fact, the extremity with the reconstructed ACL had a lower transient than the uninjured extremity. Heel strike transients in patients with a reconstructed ACL were higher than those in the controls, but the differences were significant only when corrected for velocity of gait.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410471 TI - Repair of articular cartilage injury following intra-articular chymopapain induced matrix proteoglycan loss. AB - The intra-articular injection of 0.02, 0.2, or 2.0 mg of chymopapain (CP) into the knee of adolescent rabbits caused the loss of more than 50% of the proteoglycans (PGs) in the cartilaginous tissues within the joint. Sequential measurements of cartilage-derived keratan sulfate epitope in serum and analyses of articular cartilage slices 2 days after the injection revealed that 0.02 mg of CP was nearly as effective as higher doses (0.2 or 2.0 mg of CP) in causing the depletion. The degradation and depletion of PGs in articular cartilage were shown to be localized to the joint and did not affect articular cartilage in the contralateral knee joint (no injection) or other cartilaginous tissues in the body. On day 9, partial replenishment of the articular cartilage PGs had occurred, irrespective of the dose used, and the articular surface within the joint remained intact. However, by day 21, articular cartilage in joints injected with 2.0 mg of CP had begun to show progressive degenerative changes, and these changes became more severe with time. In contrast, joints injected with 0.2 mg of CP continued to repair successfully by the reestablishment of a matrix that retained its integrity and appeared to remain functional for at least 6 months. These results suggest that the model may prove useful for the study of the repair processes that follow matrix injury and severe depletion of PGs from the articular cartilage matrix. PMID- 8410472 TI - Effect of compressive loading and unloading on the synthesis of total protein, proteoglycan, and fibronectin by canine cartilage explants. AB - Full-thickness canine articular cartilage explants were subjected to compressive loads equivalent to a uniaxial stress of 0.025-1.2 MPa. A single cycle (18 h) of unconfined compression resulted in inhibition of total protein, proteoglycan, and fibronectin synthesis. The inhibition of fibronectin synthesis followed that of total protein synthesis. The magnitude of inhibition increased nonlinearly with increasing load levels. The signal that depressed synthesis remained effective for several hours after removal of load, but by 24 h proteoglycan synthesis had partially recovered and fibronectin and protein synthesis had fully recovered and sometimes exceeded the rate of synthesis in free-swelling controls. Forty-eight hours after five cycles of intermittent unconfined compression with similar loads, proteoglycan content and synthesis did not differ in loaded disks and in disks that were never loaded in vitro. Interestingly, the percentage of water in disks that had never been loaded in vitro increased significantly after 10 days in culture, relative to the percentage of water in free-swelling disks on the day of harvest. Intermittent compressive loading in the range of 0.5-1.2 MPa partially prevented this increase. Our results confirmed the previously reported inhibition of biosynthesis with static loading but also suggested that exposure to intermittent compressive loading may help to maintain the normal ratio of dry to wet weight in the explant. PMID- 8410473 TI - Rib cage deformities in scoliosis: spine morphology, rib cage stiffness, and tomography imaging. AB - A computer-implemented biomechanical model of a thoracolumbar spine and deformable rib cage was used to investigate the influence of spine morphology and rib cage stiffness properties on the rib cage deformities that arise from scoliosis and to study the relationship of actual rib distortions with those seen on computed tomography (CT) scans. For the purposes of this study, it was assumed that rib cage deformities result from forces imposed on the ribs by the deforming spine. When a structurally normal rib cage was allowed to follow freely the imposition of scoliotic curves on the spine, different configurations of scoliosis led to substantial differences in the resulting rib cage deformities. Rib cage lateral offset correlated well with the Cobb angle of the scoliosis but not with the apical vertebral axial rotation, whereas rib cage axial rotation correlated well with apical vertebral axial rotation but not with the Cobb angle. These model-obtained findings mirror clinical findings that correction of the Cobb angle leads to correction of the lateral offset of the rib cage but does not correlate well with correction of the rib cage axial rotation. The stiffnesses of the ligamentous tissue connecting the sternum to the pelvis, of the costovertebral joints, and of the ribs themselves also influenced the rib deformities substantially. The influence of the sternopelvic ligamentous ties has not been recognized previously. The total rib cage volume remained essentially constant regardless of the severity of the resulting deformity, but the distribution of this volume between convex and concave sides varied somewhat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410474 TI - Proteoglycan synthesis in canine intervertebral discs after long-distance running training. AB - The alterations and distribution of proteoglycan (PG) synthesis in the intervertebral discs of young dogs exercised with long-distance running (40 km/day) were studied with a method based on image analysis of tissue sections. Ten dogs were run on a treadmill daily for 55 weeks, and 10 dogs from the same litters served as controls. The daily running distance gradually was increased to 40 km and was maintained at that level for the final 15 weeks. Midsagittal disc segments C7-T1, T8-9, and L1-2 were labeled with 35SO4, and histological sections of the segments were apposed against autoradiographic film to determine the synthesis of PGs. Next, the same sections were stained with safranin O to estimate possible alterations in PG concentration. The radiographs and stained sections were digitized with a flatbed scanner and measured by image analysis. The lumbar discs of runners displayed a significantly lower rate of 35SO4 incorporation, while a tendency toward enhanced incorporation was seen in the cervical and thoracic discs. Safranin O staining showed a pattern just opposite to 35SO4 incorporation: decreased staining in the cervical and thoracic discs and increased staining in the lumbar discs of the runners. The results demonstrate qualitatively different influences of long-term running training on PG metabolism at the cervical, thoracic, and lumbar levels in young dogs. PMID- 8410475 TI - The cartilage end-plate and intervertebral disc in scoliosis: calcification and other sequelae. AB - The morphology and composition of the intervertebral disc and also of the cartilage end-plate were studied in patients with idiopathic or congenital scoliosis. The cartilage end-plate was investigated because of its function as an epiphyseal plate in humans and the association between growth and progression of the scoliotic curve. The proteoglycan and water contents were reduced in both structures in specimens from scoliotic patients, particularly toward the concavity of the curve, compared with autopsy material. The distribution of some collagen types differed in tissue from scoliotic patients and autopsy tissue. Calcification of the cartilage end-plate, and sometimes of the adjacent disc, occurred in all but three scoliotic patients, whereas there was minimal calcification in the autopsy specimens. We suggest that, although these changes are probably a secondary response to altered loading in the scoliotic patients, they may be highly significant to the progression of the scoliotic curve. PMID- 8410476 TI - Stem curvature and load angle influence the initial relative bone-implant motion of cementless femoral stems. AB - A 6 df measurement system was used to investigate the initial relative bone implant motion of two types of cementless total hip replacement femoral components-a straight stem and a curved stem. Five pairs of fresh frozen femurs from human cadavers were tested with loads applied to the femoral head at angles characteristic of level walking, stair-climbing, and rising from a chair. The most important findings were that (a) the resultant proximal translations were twice as high with the straight stem as with the curved stem at load angles encountered in stair-climbing and rising from a chair, (b) both stem types had more motion at load angles encountered in stair-climbing and rising from a chair than in level walking, with the increases ranging from 3 to 14-fold, (c) there was as much as 66-fold more motion distally than proximally, and (d) the amount of interface motion varied by 5-fold between the medial and lateral surfaces of the distal part of the implant because of the combined effects of translation and rotation. The amount of initial bone-implant motion of the femoral component was found to be particularly sensitive to off-axis loading; this suggests that stair climbing and rising from a chair should be avoided in the early postoperative period when a cementless porous-coated femoral stem has been used. PMID- 8410477 TI - Chief executive officer's report. PMID- 8410478 TI - Presidential address: measures of success. PMID- 8410479 TI - Presidential report: caring, competency, and certification: the hallmarks of intravenous nurses. PMID- 8410480 TI - Levels of performance for intravenous nursing practice. AB - Intravenous nurses function at different levels of performance based on their experience, educational background, desire for autonomy, acceptance of accountability and responsibility, and commitment. Levels of practice for the IV nurse have been developed to delineate competency, proficiency, and expertise. These levels can be used to define areas of weakness, to complement achievement, and to inspire IV nurses to reach for higher goals in their professional practice. PMID- 8410481 TI - The use of confirming x-rays to verify tip position for peripherally inserted catheters. AB - The tips of long-line peripherally inserted catheters may be positioned in the superior vena cava (SVC), subclavian vein, or axillary vein. Based on current standards of practice, catheters with tips placed in the SVC are central lines that require a confirming x-ray; catheters with tips terminating in the subclavian or axillary vein are noncentral and do not require confirming x-rays. Several authors follow this standard; others, however, suggest that radiographic examination should be performed on catheters with tips lying in locations other than SVC unless numerous specific criteria are met. In this study, x-rays were obtained for 42 patients in a hospital setting to determine tip location and position. Radiographic examination revealed that six were looped and five were positioned in a branch or tributary off the axillary vein. To eliminate the complications associated with catheter tip malposition (i.e., thrombus, occlusion), x-ray confirmation should be performed on all lines placed in or beyond the axillary vein to ensure correct tip position. PMID- 8410482 TI - A clinical productivity management system for home infusion therapy. AB - Although many organizations have begun to accept the reality of cost containment in home infusion therapy, few have taken steps to cost manage their business. Even fewer home infusion providers have considered that productivity and quality will carry equal weight in the equation for providing health care and services. Organizations that address the issues of today and tomorrow's health care environment, finding a fine balance for managing productivity while providing quality care, will be those organizations that remain viable in the future. PMID- 8410483 TI - Rapid colorimetric detection of epidermal growth factor receptor mRNA by in situ hybridization. AB - We describe a rapid, formamide-free, random oligomer-enhanced in situ hybridization method in formalin-fixed, paraffin-embedded tissue sections using a biotinylated oligonucleotide probe for colorimetric detection of the mRNA transcript of the epidermal growth factor receptor (EGFR) gene, a putative protooncogene. Transitional cell carcinomas (TCC) of the urinary bladder and oral squamous cell carcinomas (SCC) were examined. A431, a human squamous carcinoma cell line that overexpresses EGFR mRNA, and mature skeletal muscle, known to express EGFR, served as control tissues. A biotinylated poly-T oligonucleotide probe was used to evaluate the preservation of mRNA in the formalin-fixed, paraffin-embedded tissues. The EGFR mRNA and poly-T oligonucleotide probes were constructed with a target-specific, 5' region and a 3' non-hybridizing, biotinylated tail. Random sequence oligomers nine bases long added to the probe cocktail eliminated the need for formamide and sheared carrier DNA in the method. The assay produced strong, specific staining for EGFR mRNA in all cases of TCC and SCC, and in the A431 tumors, but not in skeletal muscle. The assay was completed in approximately 90 minutes. This method may have widespread utility for rapid and specific detection of other mRNA sequences. PMID- 8410484 TI - Advanced glycosylation end products: a new disease marker for diabetes and aging. AB - Advanced glycosylation end products (AGEs) are a potentially useful marker for monitoring glycemic control, predicting the risk of diabetes- and aging associated clinical complications, and monitoring the treatment of patients with micro- and macrovascular diseases, including retinopathy, atherosclerosis, nephropathy, and neuropathy. AGEs or AGE-proteins are derived from nonenzymatically glycated proteins (Amadori products) after further cross-linking with other proteins and additional rearrangement. AGE-proteins can be assayed by either radioreceptor or immunoassays in blood and tissues. No commercial kit is available at this time. PMID- 8410485 TI - Immunoquantification of lipoprotein(a): comparison of nephelometry with electroimmunodiffusion. AB - A new fully automated nephelometric immunoassay for lipoprotein(a) quantification in human serum was evaluated using the Behring Nephelometer Analyzer. The assay exhibited a good linearity in the concentration range of 110-1,770 mg/l; at higher concentrations, samples were automatically diluted by a factor of 4. The method is simple, robust, and shows an excellent stability of the calibration curve over several weeks. Intra-assay and day-to-day coefficients of variation were 2% and 4.5%, respectively. The method correlated well with electroimmunodiffusion (r = 0.977; n = 123; P = 0.0001). Unspecific turbidity as expressed by an elevated blank value occurred in 3% of all freshly measured samples (n = 392). Storage of the samples for 1 week at 4 degrees C had no significant influence on the results. Frozen sera, on the other hand, cannot be assayed by this method. We believe that this assay is well suited for use in clinical routine work. PMID- 8410486 TI - Measurement of antimicrobial agents in cerebrospinal fluid using the Abbott TDx analyzer. AB - Occasionally, requests are made by our physicians for the measurement of gentamicin, tobramycin, or vancomycin in cerebrospinal fluid (CSF) specimens during the course of treating patients for bacterial meningitis. We evaluated CSF as a specimen type for the measurement of amikacin, gentamicin, tobramycin, and vancomycin on the Abbott TDx analyzer. Coefficients of variation for CSF spiked with these antimicrobial agents ranged from 0.8% to 6.5% for intra-assay values and from 2.1% to 2.3% for inter-assay values. Serum and CSF specimens were spiked at various levels with equal amounts of the antibiotics. Correlation coefficients for serum vs. CSF for these agents were 0.999. Recoveries ranged from 86% to 134%. Sensitivity for these assays is about fourfold better for CSF than for serum. CSF appears to be an acceptable specimen type for the measurement of these antibiotics using the Abbott TDx analyzer. PMID- 8410487 TI - Why a new international reference preparation (IRP) for human plasma proteins? PMID- 8410488 TI - Modern aspects and requirements for the standardization of immunoassays for human plasma proteins. AB - Some fundamental problems are summarized for an optimal standardization of protein determination in immunoassays. The relationship between a measurement signal in an immunoassay and the protein value followed by that is complicated and is explained on the basis of two immunochemical postulates and of two nonimmunochemical influences for which four postulates are derived. PMID- 8410489 TI - Review of diabetes: identification of markers for early detection, glycemic control, and monitoring clinical complications. AB - The hallmark of diabetes mellitus, whether type I or type II, is hyperglycemia. Clinical complications associated with diabetes are most likely the consequence of hyperglycemia via both altered metabolic pathways and nonenzymatic glycation of proteins. The nonenzymatic glycation of proteins is accelerated in diabetes due to elevated blood glucose concentration. The Amadori product of nonenzymatic glycation will further cross-link with other proteins to form advanced glycosylation end products (AGEs). The reaction of AGEs with long-lived proteins, such as collagen, and the uptake of AGEs by the receptors on macrophages, endothelial cells, and platelets are major reasons for the development of various clinical complications in diabetes. Several markers have been identified for the screening, diagnosis, and monitoring of the disease. Autoantibodies against beta cells are the best markers for mass screening and for early detection of type I diabetes. In addition to glycated hemoglobin, AGEs and blood glycated proteins of various half-lives could be used for monitoring glycemic control. Several abnormal metabolites have been identified as potential markers for monitoring the severity of various clinical complications. The most interesting findings in diabetic markers could be AGEs. The amount of AGEs found in the tissues could be related to the extent of micro- and macrovascular damage and might prove useful for monitoring the treatment of patients at early stages of either nephropathy, atherosclerosis, retinopathy, or neuropathy. PMID- 8410490 TI - Amyloids and tactoids. PMID- 8410491 TI - Of faddism, toxic oil, and scleroderma. PMID- 8410492 TI - Monoclonal antibodies to cytoskeletal proteins: an immunohistochemical investigation of human colon cancer. AB - Monoclonal antibodies raised to a number of microfilament-associated proteins were shown to recognize the appropriate proteins in extracts from human colon tissue. They were then used in an immunohistochemical study of normal colonic mucosa, adenomas, and adenocarcinomas. A strong reaction was seen in stromal cells within the tumours (both adenomas and adenocarcinomas) when frozen sections were stained with antibodies to filamin and caldesmon. In addition, a similar reaction was seen in the adenocarcinomas when stained with antibodies to talin and gelsolin. We believe that immunohistochemical staining with these antibodies reveals a tumour-induced process in the surrounding cells, possibly related to a host response to tumours. PMID- 8410493 TI - Isolation and characterization of multiple cell types from a single human colonic carcinoma: tumourigenicity of these cell types in a xenograft system. AB - The HCA-7 cell line was established from a moderately well differentiated mucinous adenocarcinoma of the colon, which showed histological heterogeneity. This was reflected in the morphological heterogeneity in early passages of the HCA-7 cell line but diminished as the cells were passaged in vitro. Nine subpopulations were isolated from early passage cultures of the HCA-7 line and maintained as cell lines. Each subpopulation demonstrated a unique set of stable biological characteristics in vitro. When established in vivo, there was a wide variation in xenograft generation time. The parent cell line gave rise to six distinct xenograft patterns, two of which had been observed in the primary carcinoma. Individual subpopulations yielded characteristic tumours which were composed of between one and four organizational patterns. Xenografts differed in both the organization of cells and the cell differentiation within the different patterns. Mucous cells were absent from some tumours while abundant in others. The subpopulations isolated from the HCA-7 cell line provide a new and extensive model system for studying the generation and maintenance of phenotypic heterogeneity in colorectal carcinoma. PMID- 8410494 TI - The behaviour of liposarcoma in tissue culture. A specific growth pattern. AB - There have been few studies of liposarcoma in tissue culture. We report a study of 27 cases of liposarcoma including examples of all subtypes in tissue culture. All the cases showed a uniform growth pattern of glass adherence by polygonal cells with short processes distributed in a random fashion. The cytoplasm of these cells became progressively loaded with glycogen followed by lipid droplets. These lipid droplets tended to fuse progressively to form a single or dominant vacuole. At the same time, the cells tended to adopt a rounded shape. This pattern of growth, which was seen in all subtypes of liposarcoma with minimal variation, is different from that shown by other soft tissue tumours and is similar to the growth of embryonal fat in tissue culture. PMID- 8410495 TI - P53 overexpression and Epstein-Barr virus infection in undifferentiated and squamous cell nasopharyngeal carcinomas. AB - We have analysed 22 nasopharyngeal carcinomas (NPCs) for expression of the small nuclear Epstein-Barr virus (EBV)-encoded RNAs (EBERs) and for immunohistologically detectable overexpression of p53. In situ hybridization demonstrated expression of the EBERs in 13 undifferentiated NPCs while nine squamous cell NPCs were EBER-negative. These results therefore confirm our previous DNA-DNA in situ hybridization studies and demonstrate that in the nasopharynx EBV is exclusively associated with undifferentiated but not with squamous cell carcinomas. p53 overexpression was demonstrated by immunohistology in 5 of 9 squamous cell NPCs and in 9 of 13 undifferentiated NPCs. Thus, there appears to be no correlation of p53 overexpression with EBV infection. These results are unexpected in the light of previous studies demonstrating that the p53 gene in primary undifferentiated NPC is consistently in the wild-type configuration. By contrast, analyses of squamous cell carcinomas of the head and neck have demonstrated that p53 overexpression in these cases is the result of p53 gene mutation. Whilst more detailed genetic analysis is required, our results suggest that mechanisms other than mutation of the p53 gene may be responsible for the stabilization of the protein in cases of undifferentiated NPC. It is tempting to speculate that an EBV-encoded protein may be involved. PMID- 8410496 TI - Cathepsin D immunocytochemical assays in breast carcinomas: image analysis and correlation to prognostic factors. AB - Immunocytochemical assays of cathepsin D were assessed in a series of breast carcinomas (n = 257) using monoclonal M1G8 anti-total cathepsin D and the avidin biotin-peroxidase complex. Cathepsin immunoreactivity was compared in frozen and paraffin sections. All tumours were anti-cathepsin-positive. Positive staining was observed in carcinoma and stromal cells and in the extracellular matrix. The amount of immunodetectable cathepsin in tissue was measured by computer-assisted image analysis (SAMBA 2005). Both the percentage of immunostained tumour surface and the mean optical densities were processed as continuous variables for statistical analysis and correlated with prognostic factors. It was shown that cathepsin D was independent of the tumour size, the lymph node status, hormone receptors, and pHER-2/neu overexpression. Cathepsin was significantly correlated with anti-EGFR (P = 0.012) and Ki67 (P = 0.002) immunoreactivity, tumour grade (P = 0.032), vascular invasion (P = 0.0081), proliferation index (P = 0.0045), and, to a lesser extent with AgNORs (P = 0.0504) and the degree of hyperploidy (P = 0.057). Tissue fixation and paraffin embedding significantly decreased cathepsin immunoreactivity. These results show that cathepsin D is not a totally independent prognostic factor in breast carcinomas. PMID- 8410497 TI - Quantitation of the renal arterial tree by fractal analysis. AB - To determine whether the renal arterial system has a fractal structure, the fractal dimension of renal angiograms from 52 necropsy cases was measured using an implementation of the box-counting method on an image analysis system. The method was validated using objects with known fractal dimensions. The method was accurate with errors of less than 1.5 per cent and reproducible with initial values within 1.2 per cent of the mean of ten sets of measurements (reliability coefficient 0.968, 95 per cent confidence limits 0.911-0.984). In the 36 satisfactory angiograms the mean fractal dimension was 1.61 (SD 0.06), which was significantly greater than the topological dimension of 1 (P < 0.0001), indicating that the renal arterial tree has a fractal structure. There was no significant relationship between age (P = 0.494), sex (P = 0.136), or systolic (P = 0.069) or diastolic (P = 0.990) blood pressure, but two congenitally abnormal kidneys (hypoplastic dysplasia and renal artery stenosis) had fractal dimensions at the lower end of the normal range (third percentile). Since the renal arterial tree has a fractal structure, Euclidean geometric measurements, such as area and boundary length, are invalid outside precisely defined conditions of magnification and resolution. PMID- 8410498 TI - The earliest histopathological response to hypobaric hypoxia in rabbits in the Rifugio Torino (3370 M) on Monte Bianco. AB - Twelve Dutch rabbits were kept on Monte Bianco at an altitude of 3370 m. Half of the animals were killed after 3 months, the remainder after 6 months, and a further six animals maintained at sea-level acted as controls. The carotid bodies of all the rabbits were processed for light and electron microscopy and examined qualitatively and quantitatively. The lungs were processed for light microscopical assessment of small pulmonary arterial vessels; the thickness of the pulmonary trunks and aortas were measured; and the hearts were dissected to obtain ratios of the ventricular weight. There was a slight increase in the right ventricular weight in the hypoxic rabbits but no change in the thickness of the pulmonary trunk compared with that of the aorta. In particular, there was no hypoxic remodelling of the pulmonary vasculature such as muscularization of pulmonary arterioles or intimal longitudinal muscle in pulmonary arteries. The earliest histopathological response to hypoxia occurred in the carotid bodies in the form of an increase in the count of the dark variant of chief cell after 3 months which returned to normal after 6 months. It is concluded that the carotid body of the rabbit responds with a change in its population of dark chief cells to a level of hypoxia which is insufficient to affect the pulmonary arterioles. Changes in the cardiopulmonary system can no longer be considered to be the earliest histopathological response to hypobaric hypoxia. PMID- 8410499 TI - Pathological Society of Great Britain, 167th meeting. Edinburgh, 7-9 July 1993. Abstracts. PMID- 8410500 TI - Sepsis syndrome and septic shock in pediatrics: current concepts of terminology, pathophysiology, and management. PMID- 8410501 TI - Severe endocrine and nonendocrine manifestations of the McCune-Albright syndrome associated with activating mutations of stimulatory G protein GS. AB - McCune-Albright syndrome (MCAS) is a sporadic disease classically including polyostotic fibrous dysplasia, cafe au lait spots, sexual precocity, and other hyperfunctional endocrinopathies. An activating missense mutation in the gene for the alpha subunit of GS, the G protein that stimulates cyclic adenosine monophosphate formation, has been reported to be present in these patients. The mutation is found in variable abundance in different affected endocrine and nonendocrine tissues, consistent with the mosaic distribution of abnormal cells generated by a somatic cell mutation early in embryogenesis. We describe three patients with MCAS who had profound endocrine and nonendocrine disease and who died in childhood. Two of the patients were severely ill neonates whose complex symptoms did not immediately suggest MCAS. A mutation of residue Arg201 of GS alpha was found in affected tissues from all three children. A review of the literature and unpublished case histories emphasizes the existence of other patients with severe and unusual clinical manifestations. We conclude that the manifestations of MCAS are more extensive than is generally appreciated, and may include hepatobiliary disease, cardiac disease, other nonendocrine abnormalities, and sudden or premature death. PMID- 8410502 TI - Lipoprotein(a) and apolipoproteins B and A-1 in children and coronary vascular events in their grandparents. AB - Because premature coronary vascular disease in a first-degree relative increases risk of the disease and the mechanisms may include genetically determined abnormal levels of circulating apolipoproteins, we explored the relationships between schoolchildren's apolipoprotein levels and coronary events in their parents and grandparents. We measured capillary blood concentrations of lipoprotein(a) (Lp(a) and apolipoproteins (apo B and apo A-1) in dried blood spot samples obtained by finger prick from 2010 schoolchildren aged 8 to 12 years, and questioned parents about coronary vascular events in the children's parents and grandparents. Of the 2010 questionnaires sent, 1030 (51%) were returned fully completed. Twenty-three fathers, one mother, and 645 grandparents had had coronary vascular events. There were significant associations between increased Lp(a) levels in children and the numbers of grandparents with coronary vascular events and with increasing grandparent coronary history scores (p < 0.01). There were also positive associations for apo B (p < 0.01) but none for apo A-1. Discriminate analysis showed that the log-transformed Lp(a) level was the variable most predictive of event numbers and of history scores in grandparents (Wilks lambda value = 0.984; p = 0.026); the apo B level was also predictive (Wilks lambda value = 0.988; p = 0.041), but neither the apo A-1 level nor the apo B/A-1 ratio was. We conclude that high Lp(a) and apo B levels in children aged 8 to 12 years are associated with increased risk of coronary vascular disease in older family members, even with a generation gap. These apolipoproteins may largely account for the independent contribution of family history to disease risk. Measurements of Lp(a) and apo B in schoolchildren may help to identify children and their families at increased risk and may facilitate targeting of prevention. PMID- 8410503 TI - No resuscitation and withdrawal of therapy in a neonatal and a pediatric intensive care unit in Canada. AB - STUDY OBJECTIVE: To compare and contrast the modes of death in a neonatal (NICU) and a pediatric (PICU) intensive care unit. DESIGN: Retrospective analysis of patient records. SUBJECTS: All newborn infants and children (< 17 years of age) who died in the NICU and PICU at the University of Alberta Hospitals, Edmonton, between Jan. 1, 1990, to Dec. 31, 1991. RESULTS: The mortality rate in the PICU was 8.7% (73/839) compared with 5.6% (75/1333) in the NICU (p = 0.007). Withdrawal of therapy was the most common cause of death in both units and occurred more commonly in the NICU (NICU = 69% vs PICU = 34%; p = 0.01). There were significantly more deaths as a result of failed cardiopulmonary resuscitation (CPR) in the PICU than in the NICU (29% vs 13%; p = 0.046). Death after no-CPR orders occurred with equal frequency in both units (NICU 17%; PICU 15%). Brain death accounted for 22% (16/87) of PICU deaths; no infant in the NICU was declared brain dead (p < 0.05). When deaths resulting from brain death and failed CPR were excluded, there was no significant difference between the two units regarding withdrawal of therapy (NICU 80% vs PICU 69%) and no-CPR orders (NICU 20% vs PICU 30%). CONCLUSIONS: This study confirms that both withdrawal of therapy and no-CPR orders are part of current clinical practice in both the NICU and PICU settings. The ethical foundations and implications of these practices need further elaboration. PMID- 8410504 TI - Attention-deficit hyperactivity disorder and thyroid function. AB - Attention-deficit hyperactivity disorder (ADHD) is thought to have a biologic basis, but the precise cause is unknown. It is one of the neurodevelopmental abnormalities frequently observed in children with generalized resistance to thyroid hormone (GRTH), suggesting that thyroid abnormalities may be related to ADHD. We report a prospective screening study for thyroid abnormalities in 277 children with ADHD by measurement of serum levels of total thyroxine, free thyroxine index, and thyrotropin. Fourteen children with ADHD had thyroid function test abnormalities: six had a normal free thyroxine index and elevated thyroxine level (group 1); three had a high free thyroxine index and a normal thyrotropin level (group 2); and five had a low free thyroxine index with a normal thyrotropin level (group 3). GRTH could not be demonstrated in a detailed study of four of the subjects in whom it was suspected (groups 1 and 2). Although the prevalence of ADHD in subjects with GRTH has been reported to be 46%, the overall prevalence of GRTH must be less than 1:2500 because we failed to detect GRTH in the 277 children with ADHD studied. We conclude that the prevalence of thyroid abnormalities is higher (5.4%) in children with ADHD than in the normal population (< 1%). PMID- 8410505 TI - Final attained height in patients successfully treated for childhood acute lymphoblastic leukemia. AB - We evaluated final adult height in 109 patients treated for childhood acute lymphoblastic leukemia on two multiarm Pediatric Oncology Group protocols between 1974 and 1981. Fifty-one patients received 2400 cGy cranial irradiation (XRT), and 58 patients received no XRT. All patients had no central nervous system involvement at diagnosis, achieved and maintained a complete response, entered puberty spontaneously, and had achieved final height. Height data were converted to standardized deviation scores. Mean age at diagnosis was 7.8 +/- 4.2 years. Distribution of heights at diagnosis was similar to that of the U.S. population. Relative to gender-specific heights for the population, female subjects in this study had lower attained heights than male subjects (p = 0.03). There was a monotonic trend of patients treated at an earlier age to have a reduction in final height (p = 0.057). Cranial irradiation was strongly associated with final height (mean standardized deviation score with XRT = -1.04 and without XRT = 0.14; p < 0.001). Final height was not associated with age at diagnosis, prognostic risk group, or Pediatric Oncology Group protocol. By multivariate analysis, only XRT and XRT x age were significantly associated with final height (p < 0.001 and p = 0.002, respectively). There was no significant gender effect. We conclude that XRT is significantly associated with reduced final adult height after treatment for childhood acute lymphoblastic leukemia. For survivors, therapy devoid of XRT does not appear to affect final height. PMID- 8410506 TI - Effects of passive smoking on respiratory illness from birth to age eighteen months, in Shanghai, People's Republic of China. AB - The objective of this study was to evaluate the effects of exposure to nonmaternal, passive household cigarette smoke on the incidence of respiratory illness (bronchitis and pneumonia) among children from birth through age 18 months in the Lu-Wan District, Shanghai City, People's Republic of China. A secondary objective was to evaluate the effects of other environmental factors, such as the fuel used for cooking and whether the child was breast fed, on the incidence of respiratory illness. The total daily cigarette consumption of family members was used to estimate exposure to passive smoke. The relative risks of exposure to passive cigarette smoke on the incidence of respiratory illness were 1.3, 1.7, and 2.0 for children living in households with members who smoked 1 to 9, 10 to 19, and 20 to 39 cigarettes per day, respectively, compared with the risks for children living in nonsmoking households (p for trend = 0.0002). These effects did not change materially when potential confounding factors were controlled. Children who were not fed human milk had a 1.8-fold increased risk of respiratory disease at each level of exposure to passive cigarette smoke evaluated, in comparison with children who were fed human milk for at least 1 month. PMID- 8410507 TI - Prevalence of hypertension in children with primary vesicoureteral reflux. AB - This study was undertaken to determine the prevalence of hypertension in children with primary, uncomplicated vesicoureteral reflux (VUR) and to evaluate the relationship between blood pressure (BP), grade and duration of reflux, and renal scarring. Subjects were identified retrospectively during a 17-year period; of 146 subjects who agreed to participate, 129 (88.4%) were female. Mean age at diagnosis was 5.0 years (range, 1 month to 16 years), and at follow-up was 14.4 years (range, 5 months to 21 years). Mean duration of follow-up was 9.6 years. Renal scarring was detected in 34.3% of patients by intravenous pyelogram, ultrasonography, or both. The BP at diagnosis was linearly related to the grade of reflux, but values were not higher than expected norms for age. At follow-up, mean systolic and diastolic BP were at the 41.6 percentile and the 18.7 percentile, respectively. No patient's BP was above the 55th percentile. After a mean follow-up period of 10 years, we conclude that primary, uncomplicated VUR, regardless of the number of documented urinary tract infections, duration and severity of reflux, modality of therapy, presence of renal scarring, and duration of follow-up, is not associated with the development of hypertension. Hypertension does not appear to be a complication of VUR and urinary tract infection unless there is preexisting dysplasia. PMID- 8410508 TI - Severe combined immunodeficiency: a retrospective single-center study of clinical presentation and outcome in 117 patients. AB - We carried out a retrospective analysis of 117 patients with severe combined immunodeficiency who were examined in a single center between Jan. 1, 1970, and Jan. 1, 1992, for the purpose of evaluating disease onset, progression, and outcome. The frequency of case referral increased from 8 from 1970 to 1975 to 56 from 1986 to 1991. The most frequent phenotype was T-/B+ (absence of T lymphocytes and presence of B lymphocytes) (n = 51); there were 36 cases of alymphocytosis, 16 of adenosine deaminase deficiency, 13 of Omenn syndrome, and 1 of reticular dysgenesis. Protracted diarrhea and lung infections were the main infectious complications; infection with bacillus Calmette-Guerin occurred in 10 of 28 vaccinated patients, but none of the six recipients of oral polio vaccine subsequently had poliomyelitis. The presence of maternal T cells was suspected or proved in half the patients with alymphocytosis or T-B+ severe combined immunodeficiency but did not occur in the other forms of the disease. Of the 117 patients, 22 died before transplantation could be performed. Adenosine deaminase deficiency and Omenn syndrome were more frequently associated with death before hematopoietic stem cell transplantation was possible. Fetal liver transplantation was successful in 1 of 10 cases. The survival rate among the 30 recipients of bone marrow with identical human leukocyte antigens (HLA) was 80%, with a median follow-up of 129 months; 23 of 25 patients recovered full immune function. The survival rate among the 50 recipients of HLA-haploidentical T cell-depleted bone marrow was 56%, with a mean follow-up of 35 months. Of the latter patients, 10 (35%) still require immunoglobulin substitution. There has been a trend toward improvement in the survival rate of haploidentical bone marrow recipients, presumably because of more effective infection-control measures and better transplantation strategy. PMID- 8410509 TI - The enigma of persistent hyperinsulinemic hypoglycemia of infancy. PMID- 8410510 TI - Severe chickenpox after intranasal use of corticosteroids. AB - Two children were hospitalized for severe chickenpox after intranasal use of corticosteroids for chronic sinusitis. One had unusually extensive cutaneous disease with delayed progression of lesions, dehydration, and prolonged fever; the other had hemorrhagic cutaneous lesions, hepatitis, and pneumonitis. Both were treated with intravenous acyclovir infusion and recovered. Systemic or local immunosuppression after intranasal corticosteroid administration may have predisposed the children to severe varicella infection. PMID- 8410511 TI - Effect of human immunodeficiency virus infection on the growth of young children. Duke Pediatric AIDS Clinical Trials Unit. AB - We retrospectively analyzed the growth of 170 children less than 25 1/2 months of age who were referred for evaluation of human immunodeficiency virus (HIV) antibody status. By the age of 4 months, the 62 HIV-infected children were significantly smaller than the 108 uninfected children in both weight-for-age and length-for-age measurements; linear growth and weight gain were proportionally decreased. PMID- 8410512 TI - Selection of a precore mutant of hepatitis B virus and reactivation of chronic hepatitis B acquired in childhood. AB - A 14-year-old girl with chronic hepatitis B had seroconversion from hepatitis B e antigen to antibody and achieved biochemical remission after 2 years. The disease reactivated 9 years later when a precore mutant had become the prevalent hepatitis B virus strain in serum. These results suggest that selection of a precore mutant might induce reactivation during adult life of chronic hepatitis B acquired in childhood, thus worsening the prognosis. PMID- 8410513 TI - Rheumatic symptoms associated with hypothyroidism in children. AB - We describe five children with varied rheumatic manifestations, including fibromyalgia and arthralgias, ultimately proved to be associated with hypothyroidism. All musculoskeletal symptoms improved after thyroid replacement therapy. We conclude that rheumatic manifestations of hypothyroidism can be as varied in children as in adults. PMID- 8410514 TI - Benign familial infantile epilepsy. AB - We report 23 subjects with afebrile seizures. The subjects, from 11 families, were between the ages of 3 and 19 months and had normal neurodevelopmental status, a normal interictal electroencephalogram, a family history of similar seizures, and an excellent prognosis. Benign familial infantile epilepsy is a clinical syndrome that may be more common than is presently recognized. PMID- 8410515 TI - Infant feeding and idiopathic intussusception. AB - A case-control study showed that, compared with infants who had never been fed human milk, breast-fed infants had a relative risk of intussusception of 6.0 (95% confidence interval, 1.8 to 20.4) when breast-feeding at admission was exclusive and of 2.3 (95% confidence interval, 0.8 to 6.6) when it was partial. Exclusive breast-feeding may be a risk factor for intussusception in infancy. PMID- 8410516 TI - Molybdenum cofactor deficiency. AB - We describe a new case of molybdenum cofactor deficiency, an underrecognized inborn error of metabolism that results in neonatal seizures and neurologic abnormalities. Characteristic biochemical defects in affected individuals include hypouricemia, elevated urine sulfate (detectable by dipstick), and elevated S sulfocysteine (detectable by anion exchange chromatography). This disorder should be considered in the differential diagnosis of neonatal seizures. PMID- 8410517 TI - Identical mitochondrial DNA deletion in mother with progressive external ophthalmoplegia and son with Pearson marrow-pancreas syndrome. AB - We describe a family in which the mother has progressive external ophthalmoplegia with the common 4977 base pair deletion, and her son has a syndrome similar to the Pearson marrow-pancreas syndrome with the identical deletion. This case extends the clinical phenotype of the Pearson syndrome and raises the possibility that developmentally regulated tissue-specific nuclear factors are responsible for the differential phenotypic expression of these two mitochondrial disorders. PMID- 8410518 TI - Pulmonary hemorrhage and exogenous surfactant therapy: a metaanalysis. AB - A metaanalysis of surfactant clinical trials was carried out to assess whether or not an association exists between exogenous surfactant therapy and pulmonary hemorrhage. Trials that reported the pulmonary hemorrhage occurrence (group 1) and those that did not (group 2) were analyzed. Thirty-three treatment strategies were tested in 29 publications from 1980 through 1992. Eleven of these were group 1 trials, which reported a 3% overall incidence of pulmonary hemorrhage. The rates were significantly higher in both the treated and the control groups of natural surfactant trials than in synthetic surfactant trails (5.87% and 5.36% in the natural surfactant trials vs 2.51% and 1.04% in the synthetic surfactant trials, respectively). The pooled estimate of relative risk for pulmonary hemorrhage with any surfactant therapy was 1.47 (95% confidence interval 1.05, 2.07; p < 0.05). Logistic regression modeling revealed that the nature of surfactant, treatment strategy, and lower mean birth weight had a significant influence on the relative risk of pulmonary hemorrhage; a similar trend was seen with higher mortality rates. Variation in the rates of patent ductus arteriosus did not have an independent effect on the estimated pulmonary hemorrhage risk. Most group 2 trials were published before 1990, and the median total sample size was 73, compared with 402 for the group 1 trials (p < 0.05), most of which were published in the 1990s. In 10 (50%) of 20 group 2 trials, pulmonary hemorrhage data were collected methodically, in comparison with all group 1 trials, most of which collected data prospectively. We conclude that pulmonary hemorrhage is a rare complication of respiratory distress syndrome. An awareness of the possible association of pulmonary hemorrhage with surfactant use in later trials and the differences in definitions and reporting practices probably explain variations in the reported incidence among the trials. The risk of pulmonary hemorrhage increases slightly, on an average of 47%, with any surfactant therapy. This increased risk is small compared with the documented benefits of surfactant therapy in respiratory distress syndrome. PMID- 8410519 TI - Prognostic implications of age at detection of air leak in very low birth weight infants requiring ventilatory support. AB - STUDY OBJECTIVE: To measure the association between the development of air leak (pneumothorax or pulmonary interstitial emphysema) during the first 27 postnatal days and neonatal death or chronic lung disease, as determined on day 28, among very low birth weight infants who required mechanical ventilation from the first day of life. DESIGN: Prospective, multicenter cohort study. PATIENTS: Two hundred sixty inborn, very low birth weight (501 to 1500 gm) infants given ventilatory support from the first day of life. RESULTS: The risk of an adverse outcome (death or chronic lung disease) changed with postnatal age at the time of diagnosis of the air leak. The association between air leak and an adverse outcome, as measured by gestational age-adjusted odds ratio (95% confidence interval), was 13.9 (1.7 to 114.6) for those in whom an air leak developed on day 0 or 1 (early), decreased to 1.7 (0.7 to 4.1) for those whose air leak developed on day 2 or 3 (intermediate), and increased to 16.6 (2.1 to 130.4) for those whose air leak developed on days 4 to 27 (late). The association with neonatal death showed even more striking fluctuations with postnatal age at occurrence of an air leak, ranging from an odds ratio of 40.3 (3.5 to 464.8) for the early group to 7.5 (2.3 to 25.0) for the intermediate group and 78.3 (6.9 to 889.5) for the late group. CONCLUSIONS: Air leak in newborn infants requiring mechanical ventilation is associated with increased risks of death or future morbidity, but the magnitude of these risks changes with postnatal age at the time of diagnosis of the air leak. Failure to consider the age at which the air leak is detected may miss changes in its prognostic implications and may partly explain inconsistent results in previous studies. PMID- 8410520 TI - Early changes in the neonatal circulatory transition. AB - To define the course of neonatal circulatory transition and to identify clinically relevant echocardiographic measurements in the diagnosis of persistent pulmonary hypertension, we prospectively studied 32 healthy term infants from 30 minutes to 24 hours after birth with color and quantitative Doppler echocardiography on the first day of life, and compared them with 33 term infants supported by mechanical ventilation for respiratory failure. Color Doppler imaging included measurements of cardiac output, left pulmonary artery flow, aortopulmonary pressure difference, ductal flow, left-to-right color-flow jet area of the ductus arteriosus, and ductal flow characteristics. In healthy infants the majority of measurable changes in cardiopulmonary hemodynamics had occurred by 8 hours after birth, although some degree of right-to-left ductal shunting was found up to 12 hours after birth. In the infants with respiratory failure, ductal flow and maximum aortopulmonary pressure difference measurements at 8, 12, and 24 hours showed a significant delay in ductal closure and a high incidence of persistent pulmonary hypertension, which correlated well with the severity of their respiratory failure. Factors such as aortopulmonary pressure difference, prolonged right-to-left shunting with decreased left pulmonary artery flow, and failure to develop a left-to-right ductal color-flow jet were found to be practical markers for assessing the course of neonatal circulatory transition in sick term infants. PMID- 8410521 TI - Nitrogen balance in extremely low birth weight infants with nonoliguric hyperkalemia. AB - We measured nitrogen balance and urinary 3-methylhistidine molar ratios in 33 extremely low birth weight infants (12 with hyperkalemia and 21 without) for the first 3 days of life. Although all infants were in negative nitrogen balance during the study, there was no difference in the degree of negative nitrogen balance between the two groups. There was also no difference in the 3 methylhistidine/creatinine molar ratios, indicating that muscle protein catabolism did not differ. We conclude that it is unlikely that catabolism contributes to the development of nonoliguric hyperkalemia in extremely low birth weight infants. PMID- 8410522 TI - Short- and long-term use of octreotide in the treatment of congenital hyperinsulinism. AB - Octreotide, a long-acting analog of somatostatin that inhibits insulin release, has the potential to control hypoglycemia in infants with congenital hyperinsulinism. To examine the efficacy and side effects of octreotide, we evaluated therapy between 1988 and 1993 in 16 infants who did not respond to diazoxide. In nine patients with onset of severe hypoglycemia in the first days of life, octreotide was helpful in stabilizing plasma glucose levels and allowed reductions in the rates of glucose infusion; however, glucose control was inadequate to avoid subtotal pancreatectomy. In two of these nine patients postoperatively and in seven other infants, a trial of long-term treatment with octreotide was undertaken. Four were treated successfully for up to 4.3 years. Octreotide therapy was not associated with thyroid deficiency and caused only transient malabsorption. All patients receiving long-term therapy had some decrease in linear growth and two had subnormal plasma concentrations of insulin like growth factor I and insulin-like growth factor binding protein 3 compatible with suppression of growth hormone by octreotide. Resistance to octreotide therapy, even with increasing doses, occurred in all patients. These results suggest that octreotide may aid in the acute or long-term treatment of congenital hyperinsulinism in a limited number of selected cases. PMID- 8410523 TI - Persistent hyperinsulinemic hypoglycemia of infancy: long-term octreotide treatment without pancreatectomy. AB - Eight patients with persistent hyperinsulinemic hypoglycemia of infancy who were treated with octreotide without pancreatectomy are described. All had severe, early-onset disease that would have required partial pancreatectomy had octreotide not been available. Along with octreotide, frequent feedings and raw cornstarch at night were required by all. Octreotide was given in three or four daily subcutaneous injections in four patients and in a continuous subcutaneous infusion with an insulin infusion pump in four. All had mild, transient gastrointestinal symptoms (vomiting, abdominal distention, steatorrhea) after the start of therapy. Asymptomatic gallstones were found in 1 patient after 1 year of treatment. No other long-term untoward effects were noted, including no detrimental effect on psychomotor development. Growth was not affected in five of six patients treated for more than 6 months. In five patients, octreotide was discontinued after 9 months to 5 1/2 years; patients were given diazoxide instead, two required percutaneous gastrostomy, and one 5 1/2-year-old child required no further treatment. The remaining three patients (aged 5 to 9 months) are still being treated with octreotide. We conclude that, with the use of octreotide, pancreatectomy can be avoided in some patients. Particularly in light of our findings of a high incidence of diabetes years after partial pancreatectomy, and clinical improvement after months to years of octreotide treatment, we believe that aggressive medical therapy, when effective, is preferable to partial pancreatectomy. PMID- 8410524 TI - Gamma globulin re-treatment in Kawasaki disease. AB - We retrospectively reviewed the effects of intravenous gamma-globulin (IVGG) re treatment of 13 children with Kawasaki disease and persistent or recrudescent fever. Fever and mucocutaneous inflammation resolved within 48 hours in nine patients; fever abated in two other children after a third course of IVGG. We conclude that IVGG re-treatment of Kawasaki disease appears to be safe and may improve the clinical course. PMID- 8410525 TI - A fifteen-year-old boy with eosinophilia and pulmonary infiltrates. PMID- 8410526 TI - Prophylaxis for rheumatic fever using benzathine penicillin. PMID- 8410527 TI - Transient methemoglobinemia, diarrhea, and dietary protein intolerance. PMID- 8410528 TI - Treatment of hypertension in acrodynia. PMID- 8410529 TI - Physiopathologic mechanism of hypercalciuria in renal glucosuria. PMID- 8410530 TI - Herbal teas for infantile colic. PMID- 8410531 TI - Herbal teas for infantile colic. PMID- 8410532 TI - Gitelman versus classic Bartter syndrome. PMID- 8410533 TI - Leptomonas seymouri as a model system for the analysis of gene expression in trypanosomatids. AB - Leptomonas seymouri, a monogenetic trypanosomatid originally isolated from Dysdercus suturellus (Hemiptera), was used to develop a reverse genetic system for trypanosomatid flagellates. In many eukaryotic cell types, reverse genetics has proven to be a powerful tool for defining structure/function relationships within genes. The mini-exon genes of trypanosomatids encode key components of all cellular mRNAs. This component is a 5' "leader" RNA that is spliced onto all mRNA precursors during mRNA formation within the cell nucleus. The data presented here indicate that structure/function relationships within the mini-exon gene can be probed using the molecular genetic system developed and characterized for L. seymouri. PMID- 8410534 TI - A family of target site-specific retrotransposons interrupts spliced leader RNA genes in trypanosomatids. AB - Retrotransposons are mobile genetic elements that have been found in almost every genome. Because they do not code for any coat protein sequences, it is believed that they have lost the ability to maintain an extracellular life cycle yet they share many other gene homologies with retroviral genomes. Their replication involves an RNA intermediate and the activity of the reverse-transcriptase enzyme for which they code. Although the majority of these elements are promiscuous in their insertion, there are a few examples that integrate into specific target sites. One such integration site-specific element interrupts the spliced leader RNA genes of organisms of several genera within the family Trypanosomatidae. These elements constitute a highly related family of sequences that most likely have evolved through a common ancestor. PMID- 8410535 TI - Analysis of malarial transcripts using cDNA-directed polymerase chain reaction. AB - A polymerase chain reaction (PCR)-based approach is being employed to study RNA transcripts in malarial parasites, a system that is not easily amenable to molecular studies. Our aim is to compare messages from different life cycle stages to determine whether regulatory information is encoded in the structure of plasmodial transcripts as a result of differential RNA processing. In particular, we have analyzed the structure of the message encoding the circumsporozoite (CS) protein of the murine malaria Plasmodium berghei, the immunodominant surface antigen of the infectious stage of the parasite. Our major findings are that a subset of the CS message utilizes multiple polyadenylation sites, that some processed CS transcripts are found in blood-stage parasites, and that the 5' untranslated region of the message is unusually long and has multiple start sites. Moreover, repetitive motifs that may represent enhancers or transcriptional binding sites are present upstream of the transcription unit. In addition, we describe details of the cDNA-directed PCR procedure that may be helpful to other parasitologists who work with small, unpurified biological samples. PMID- 8410536 TI - Biogenesis of the hydrogenosome in the anaerobic protist Trichomonas vaginalis. AB - Trichomonas vaginalis is a primitive protist that lacks mitochondria and peroxisomes. This aerotolerant anaerobe does, however, contain a specialized organelle involved in carbohydrate metabolism called the hydrogenosome. Hydrogenosomes are the site of fermentative oxidation of pyruvate, coupled to ATP production via substrate level phosphorylation. The organelle is surrounded by 2 membranes and appears to contain no genetic material. Hydrogenosomes are proposed to have arisen either through the conversion of mitochondria or via endosymbiosis with an anaerobic bacterium. Our studies show that hydrogenosomal proteins are synthesized on free polyribosomes and are, thus, posttranslationally translocated into the organelle. The 2 hydrogenosomal proteins we have examined in detail, ferredoxin and the beta subunit of succinyl CoA synthetase, appear to be synthesized as larger precursors that contain an 8- or 9-amino acid amino terminal extension, respectively. These apparent leader sequences are absent from the mature proteins found in hydrogenosomes. The biochemical properties of the amino-terminal extensions are similar to one another and to mitochondrial presequences. These observations are consistent with the hypotheses that the specificity of protein translocation into hydrogenosomes is conferred by a short amino-terminal signal sequence and that the mechanisms underlying translocation are similar to that of mitochondria. Using in vitro import assays, we have shown that this short leader sequence is absolutely required for association of hydrogenosomal proteins with the organelle and that this association is dependent on ATP, temperature, and the presence of intact organelles. PMID- 8410537 TI - Buticulotrema stenauchenus n. gen., n. sp. (Digenea: Opecoelidae) from Malacocephalus occidentalis and Nezumia aequalis (Macrouridae) from the Gulf of Mexico. AB - Buticulotrema stenauchenus n. gen., n. sp. (Opecoelidae: Opecoelinae) is described from the macrourid fishes Malacocephalus occidentalis Goode and Bean, 1885, and Nezumia aequalis Gunther, 1878, collected in June 1971 from the DeSoto Canyon area of the Gulf of Mexico at a depth of 495 m. Eight of 25 (32%) M. occidentalis and 1 of 8 (13%) N. aequalis examined were infected with the new opecoelid. Specimens of Bathygadus macrops Goode and Bean, 1886, and Coelorhynchus coelorhynchus Risso, 1810, collected at the same time were not infected with B. stenauchenus. The new genus most closely resembles Genitocotyle Park, 1937, but differs in that it lacks an accessory sucker behind the genital pore, contains a blind rather than canalicular seminal receptacle, and has a slightly pedunculate acetabulum, a pharynx that is longer than wide, and a distinctive, long-necked, bottle-shaped body. Buticulotrema stenauchenus is also similar to Pseudopecoelus von Wicklen, 1946, but the latter lacks a blind seminal receptacle. PMID- 8410538 TI - Gymnophalloides seoi n. sp. (Digenea: Gymnophallidae), the first report of human infection by a gymnophallid. AB - Gymnophalloides seoi n. sp. (Digenea: Gymnophallidae) is described from worms expelled from a naturally infected women in Korea. The patient, a 66-yr-old housewife, experienced severe epigastric discomfort, and fecal examination revealed numerous parasite eggs of 0.019-0.021 x 0.014-0.016 mm. After anthelmintic treatment and purgation, 952 digeneans consisting of 910 adult gymnophallids and 42 heterophyids (2 species) were recovered. The gymnophallid flukes closely resemble Gymnophalloides tokiensis Fujita, 1925 (known only as metacercariae from oysters), type and only previously known species of the genus, except for a difference in the position and orientation of the seminal vesicle. Without experimental proof of identity, the adult gymnophallids have been described as a new species, G. seoi, the only gymnophallid known to infect humans. PMID- 8410539 TI - Immature Ixodes scapularis (Acari: Ixodidae) parasitizing lizards from the southeastern U.S.A. AB - Preserved museum specimens of 13 lizard and 3 snake species common in the southeastern U.S.A. were examined for immature Ixodes scapularis Say ticks. Five Eumeces and 4 Ophisaurus lizard species yielded an infestation prevalence of 17.8% for species of Eumeces and 29.0% for species of Ophisaurus. Mean intensity of larvae and nymphs was 7.1 and 2.7, respectively, for species of Eumeces, and 6.3 and 1.4, respectively, for species of Ophisaurus. Collection dates of the lizards ranged from January through December, but most were collected from March through October. The maximum number of immatures found on a single specimen was 193 larvae and 11 nymphs on a specimen of Eumeces and 75 larvae and 7 nymphs on a specimen of Ophisaurus. For species of Eumeces, 75.2% of all nymphs observed were attached in the shoulder area. Larvae were most abundant on the rear legs (53.3%), followed by the front legs (26.3%) and shoulders (12.9%). Larvae and nymphs on species of Ophisaurus were found almost exclusively in the lateral groove area (84.0% and 94.4%, respectively). Three other lizard species (Anolis carolinensis, Sceloporus undulatus, Scincella lateralis) had only a few ticks or none. A fourth species, Cnemidophorus sexlineatus, hosted a total of 3 larvae on 3 specimens (infestation prevalence, 10.7%). Three snake species (Diadophis punctatus, Virginia striatula, Crotalus adamanteus) had none. PMID- 8410540 TI - The metazoan parasite community of migrating greater yellowlegs, Tringa melanoleuca, from the Rio Grande Valley, Texas and New Mexico. AB - Forty-eight migrant greater yellowlegs, Tringa melanoleuca, collected from southwestern Texas and southeastern New Mexico were examined for metazoan parasites. Nine helminth and 7 ectoparasite species were collected. Five new host records were recorded. The helminth fauna showed little diversity, little concentration for dominance, and no significant positive or negative associations between species were found. All species of helminths showed a contagious distribution. The component helminth community consisted of 2 core, 3 secondary, and 4 satellite species, and there was no host specialist. A checklist of metazoan parasites reported from greater yellowlegs in North America is included. PMID- 8410541 TI - Cryopreservation of infective larvae of Onchocerca volvulus (Filarioidea: Onchocercidae). AB - Infective larvae (L3) of Onchocerca volvulus were procured in Liberia, West Africa, in the natural black fly vector, Simulium yahense. A cryobiological technique was developed to preserve L3 of O. volvulus that were fully viable after thawing. Larvae were treated before cooling with 4 cryoprotective compounds. Three compounds, dimethyl sulfoxide (DMSO), glycerol, and ethylene glycol, were prepared with distilled water. The fourth compound was DMSO prepared in different concentrations with 0.25 M sucrose. The treatment with DMSO + 0.25 M sucrose cryoprotectant resulted in the highest survival of infective larvae. Five cooling rates between 0.5 C/min and 20.0 C/min were applied. The highest survival of L3 was with the cooling rate of 1.0 C/min. Two-step cooling of L3 was applied. In the first step, L3's were frozen to 5 levels from -10.0 C to -20.0 C, -30.0 C, -40.0 C, -60 C, and -80.0 C, and in the second step, larvae were transferred into liquid nitrogen at -196 C for rapid cooling and storage. The survival was the highest when larvae were cooled to approximately -40 C prior to transfer into liquid nitrogen. Slow, gradual, and rapid thawing procedures were applied. The survival was the highest in rapid warming. PMID- 8410542 TI - Inhibition and recovery of tick functions in cattle repeatedly infested with Boophilus microplus. AB - Six tick-naive male Hereford calves were infested with about 18,000 Boophilus microplus larvae, 400 in a cloth bag, once a month for 6 mo. Working with the bag ticks, 12 tick characteristics were recorded for each infestation. Manifestations of resistance to some attributes appeared by the second infestation, peaked by the third (detachment weight, egg weight, fertility index) or fourth (feeding length, start of oviposition), and vanished by the fifth or sixth. Resistance to corpse weight, oviposition length, start of eclosion, feeding index, and survival developed more slowly but continued until the last infestation. Lymphocyte transformation after the fifth infestation was normal for concanavalin A and augmented in only 2 calves for phytohemagglutinin. In each infestation and at the end of all infestations some calves were significantly more resistant to some tick characteristics. There was no relationship between natural resistance and the ability to mount an acquired resistance. We concluded that B. microplus can depress or evade host's immunity, that resistance to different tick functions occurs independently, that natural and acquired resistance are unrelated, and that there exist genetic differences in natural or acquired resistance to the tick among seemingly homogeneous animals. PMID- 8410543 TI - Immunization of cats with tissue cysts, bradyzoites, and tachyzoites of the T-263 strain of Toxoplasma gondii. AB - Previous studies have demonstrated that oral administration to cats of tissue cysts of the oocyst-negative mutant strain of Toxoplasma gondii, T-263, induces immunity to oocyst shedding following challenge. Experiments were designed to compare the levels of protection induced by T. gondii T-263 when tissue cysts, bradyzoites released from tissue cysts, and tachyzoites were administered to cats. In 1 experiment, groups of cats received 2 oral doses of intact tissue cysts or released bradyzoites of T. gondii T-263 and were challenged 47 days later with the oocyst-producing strain of T. gondii T-265. All cats seroconverted following immunization and none of them shed oocysts following challenge. In a second experiment, groups of cats received tachyzoites of T. gondii T-263 as a single oral dose and either 1 or 2 intraduodenal doses; they were challenged 60 days after the last vaccination. All cats seroconverted following immunization. Following challenge, all cats shed oocysts except for 2 of 7 cats that received 2 intraduodenal doses of tachyzoites. Thus, orally administered bradyzoites of T. gondii T-263, either contained in intact tissue cysts or liberated from cysts, induced immunity to oocyst shedding. In contrast, tachyzoites did not completely protect against oocyst shedding, even when delivered directly to the duodenum and despite the development of high antibody titers. PMID- 8410544 TI - Host specificity of Calyptospora funduli (Apicomplexa: Calyptosporidae) in atheriniform fishes. AB - Calyptospora funduli has a broad host specificity, infecting at least 7 natural and 10 additional experimental definitive hosts, all atheriniform fishes within 5 families, but most in the genus Fundulus. Barriers, apparently innate ones, prevent any development of C. funduli in perciform fishes but allow incomplete or abnormal development of the parasite in a few unnatural atheriniform hosts. In the freshwater species Fundulus olivaceus and Fundulus notti, these abnormalities consisted of asynchronous development, degeneration of the parasite in early stages of development, and the formation of numerous macrophage aggregates. Rivulus marmoratus has the ability to eliminate infections with a granulomatous inflammatory response. Additional barriers that limit natural infections of C. funduli in other hosts include feeding behavior, environmental conditions, and geographic isolation. PMID- 8410545 TI - Migration of adult Elaphostrongylus rangiferi (Nematoda: Protostrongylidae) from the spinal subdural space to the muscles of reindeer (Rangifer tarandus). AB - To trace the intrahost migration of adult (fifth-stage) Elaphostrongylus rangiferi 9 reindeer calves were fed infective larvae and examined for worms 48 250 days postinfection. The average length of worms recovered increased asymptotically with time. Gravid females and adult males were recovered from 52 days postinfection (d.p.i.), and the fraction made up by these categories increased with time. Immature females and subadult males were recovered as late as 161 d.p.i. Nematodes were recovered from the spinal subdural space 48-161 d.p.i., from the cranial subdural space 48-90 d.p.i., and from the musculature 90 250 d.p.i. Immature females were found in the spinal subdural space and the cranial subdural space, whereas gravid females were found also in the musculature with an increasing fraction with time. Subadult and adult males were found in all 3 sites but with an increasing fraction of adults from the spinal canal to the cranium to the musculature. PMID- 8410546 TI - Enteral and parenteral phases of Trichinella nativa and Trichinella pseudospiralis in the deer mouse, Peromyscus maniculatus. AB - Trichinella nativa and Trichinella pseudospiralis infections in a wild rodent host, the deer mouse (Peromyscus maniculatus), were characterized. Forty-six percent of 400 inoculated T. nativa were recovered on day 4 postinoculation (PI); 77% and 23% were found in the small and large intestines, respectively. Thirty one percent of the worms recovered on day 4 remained in the large intestine beyond day 20 PI. Worms were embedded in the mucosa of the small intestine, cecum, and colon. Females recovered from the small and large intestines had statistically indistinguishable in vitro larval releases. Distension of the cecum and passage of loose stools were associated with the presence of worms in the large intestine. The ability of T. nativa to establish and thrive in the large intestine of deer mice was confirmed following intracecal implantation of first stage larvae. On day 4 PI, 35% of 400 inoculated T. pseudospiralis were recovered, with 91% and 9% found in the small and large intestines, respectively. Although T. pseudospiralis established in the large intestine of deer mice, few worms remained beyond day 20. Females recovered from the small and large intestines had statistically indistinguishable in vitro larval releases. Although higher establishments of T. nativa (56% vs. 46%) and T. pseudospiralis (52% vs. 35%) were observed in CD-1 mice than in deer mice on day 4 PI, neither was able to colonize the large intestine of the former. The large intestine may be a more important habitat for adult trichinae than previously recognized. PMID- 8410547 TI - Patterns of morphological variation of Salsuginus yutanensis (Monogenea: Ancyrocephalidae) over space and time. AB - Salsuginus yutanensis occurs on the gills of the plains topminnow Fundulus sciadicus Cope. The fish of this species have been found to vary morphologically and biochemically among disjunct populations. Morphological characteristics of the sclerotized parts of S. yutanensis were examined from 3 localities in Nebraska, over a 2-yr collecting period. Analysis of variance was used to assess morphological variation with respect to site and date. Worms from 2 localities, Keith and Saunders counties, differed significantly for most characters considered. A third site, also in Keith County, contained worms for which measurement means tended to be intermediate between those in the other 2 sites. This site-related difference was maintained over a pattern of broad seasonal variation and suggests that the site-related differences are of evolutionary origin. If this interpretation is true, then the parasite populations likely are isolated in a manner analogous to those of the host. However, differences due to effects of temperature on worm development were not ruled out as possible explanations for the observations although consistent temperature differences between the sites are unlikely, given the nature of the habitats studied. PMID- 8410548 TI - Evidence for involvement of endogenous opioid peptides in altered nociceptive responses of mice infected with Eimeria vermiformis. AB - Parasite modification of host behavior is a well established phenomenon; however, little is known about the modulatory mechanisms regulating such effects. This study examined the relationship between Eimeria vermiformis infection, nociceptive responses, and endogenous opioid peptide activity in male RML mice. Infected mice displayed increases in centrally mediated antinociceptive responses (i.e., analgesia, measured as the latency of a foot-lifting response to a 50 C surface) throughout the prepatent period. The level of analgesia declined following onset of patency on day 8 postinfection (PI). Opioids were implicated in the increased antinociceptive response of the infected mice as the response was blocked by administration of a prototypic opiate antagonist, naloxone (1.0 mg/kg), on day 7 PI when maximum levels of analgesia were noted. This indicates that the analgesia evident in the parasitized mice was associated with increased opioid activity. Analgesia is one of a variety of behaviors influenced by changes in opioid activity, thus these observations provide further support for the contention that other parasite-induced alterations in host behavior may, in part, be the result of alterations in the activity of opioid modulatory systems. PMID- 8410549 TI - Gross lesions and hematological changes in domesticated mallard ducklings experimentally infected with Cyathocotyle bushiensis (Digenea). AB - The digenean Cyathocotyle bushiensis has been associated with late summer fatalities of dabbling ducks in southern Quebec. The objective of this study was to investigate the intensity- and time-dependent pathogenesis induced by experimental infection with this digenean in domesticated mallard ducklings. Lesions, hemorrhagic spots, plaques, and cores were observed in the ceca. Both affected tissue area and core weight increased with increasing intensity of infection. Magnitude of tissue damage increased with increasing duration of infection. Dye, administered intravenously, was found within the cecal lumen of infected ducks demonstrating increased permeability of the cecal wall to vascular products. The data suggest that infection was associated with decreased weight gain and a minor increase in body temperature. Elevations in hemoglobin concentration and packed cell volume were observed in some infected ducklings. PMID- 8410550 TI - Induction of hepatic inflammatory response by Plasmodium berghei sporozoites protects BALB/c mice against challenge with Plasmodium yoelii sporozoites. AB - BALB/c mice are about 2,000 times less susceptible to sporozoites of Plasmodium berghei than to Plasmodium yoelii. Associated with this is the innate cellular response mounted after injection with P. berghei. Host inflammatory cells do not normally attack P. yoelii during their development as exoerythrocytic forms (EEFs) in the liver. We used P. berghei sporozoites to induce host inflammation that might act against developing P. yoelii EEFs. Mice injected with P. berghei sporozoites followed 1 hr later with P. yoelii had a 58% reduction in P. yoelii EEFs. To establish whether this was due to events that occurred before vs. after invasion of hepatocytes by P. yoelii sporozoites, mice received P. yoelii sporozoites that were allowed to invade for 1 hr before subsequent injection with P. berghei; these mice showed minimal reduction in P. yoelii EEFs. Thus, most of the deleterious effects of P. berghei sporozoites appear to have been directed against P. yoelii sporozoites prior to their invasion of hepatocytes. Plasmodium yoelii that had already invaded were relatively unaffected. Further timing experiments showed that this effect was induced only by viable P. berghei sporozoites, which may thus induce rapid changes in sinusoid physiology leading to host resistance against P. yoelii sporozoites. PMID- 8410551 TI - Efficacy of ivermectin in a beef-based chewable formulation against Ancylostoma caninum and Uncinaria stenocephala in dogs. AB - The effective dosage of a chewable formulation of ivermectin was determined in 35 young dogs with induced infections of Ancylostoma caninum and Uncinaria stenocephala. Dogs were inoculated with these parasites and held until the infections were patent. Within each of 7 replicates, dogs were allocated randomly to 1 of 5 treatment groups: vehicle control, or ivermectin at 6, 12, 18, or 24 micrograms/kg. Chewable treatments were tailored to body weight. Seven or 8 days after treatment, parasites were recovered using standard techniques. All 7 controls had adult A. caninum (geometric mean = 35.5) and U. stenocephala (geometric mean = 82.6). Against A. caninum, the efficacy of ivermectin was 52%, 98%, 95%, and 97% at 6, 12, 18, and 24 micrograms/kg, respectively. The statistical model that best described the dose response was linear to 12 micrograms/kg with a plateau thereafter. Using this model, the estimated reduction from the predicted control mean was 97.2%; the estimated dose to eliminate 90% of the worms (ED90) was 8.4 micrograms/kg, and the ED95 was 10.5 micrograms/kg. Against U. stenocephala, the dose response was linear in the range studied, with an ED90 of 20.8 micrograms/kg; it was estimated that 93.2% of the worms would be eliminated. PMID- 8410552 TI - Paromomycin is effective as prophylaxis for cryptosporidiosis in dairy calves. AB - Of 16 experimentally infected neonatal dairy calves, 12 were fed paromomycin twice daily in their milk for 11 consecutive days beginning 1 day before oral inoculation with 1.5-2.0 x 10(6) oocysts of Cryptosporidium parvum. Four calves each in groups A, B, C, and D received total daily doses of 100, 50, 25, and 0 mg of paromomycin per kilogram of body weight, respectively. From birth until 28 days of age feces from each calf were examined for diarrhea, and oocysts were enumerated, rectal temperature was recorded, and weight gain was determined. Total days of diarrhea, severity of diarrhea, the total number of days oocysts were shed, and the number of oocysts shed were significantly less in group A than in the unmedicated group D. The severity of diarrhea was also significantly less in groups B and C than in group D. Oocysts were not detected in feces from calves in group A. Except for 1 calf, oocysts were not detected from calves in groups B and C during the first week the drug was administered and those calves that shed oocysts began shedding at or near the end of paromomycin administration or more than 1 wk after treatment ended. Frequency of fever and weight gains did not vary significantly between the unmedicated and medicated groups except for group C, calves of which gained significantly less weight than those in all other groups. PMID- 8410553 TI - Prophylactic activity of tetracycline against Brugia pahangi infection in jirds (Meriones unguiculatus). AB - The ability of oral tetracycline to inhibit the development of third-stage infective larvae (L3) of Brugia pahangi to adult worms in jirds was studied using 2 experimental protocols. Jirds treated with 1.4% tetracycline in drinking water for a period beginning 30 days before inoculation of L3 until 30 days post inoculation (DPI) had 97% reduction in adult worm recovery compared to untreated controls. Jirds that received 1.2% tetracycline in drinking water beginning 1 day before until either 12 or 26 DPI had adult worm recoveries of 11% and < 1%, respectively. Untreated jirds and those given tetracycline beginning at or later than 13 DPI had similar adult worm recovery (27-29%). Prepatent periods were prolonged, and circulating microfilariae were reduced in jirds given tetracycline from 27 to 54 DPI compared to controls. These data indicate that tetracycline administered to jirds in drinking water inhibits B. pahangi development from L3 to adult worms and suggest that this effect occurs during early larval development. Tetracycline administered to infected jirds prior to and continuing through the onset of patency can also affect development of microfilaremia. PMID- 8410554 TI - Third-stage larvae emerge from eggs of Contracaecum osculatum (Nematoda, Anisakidae). AB - In Contracaecum osculatum Rudolphi, 1802 (Nematoda, Anisakidae) the first and the second ecdyses occur inside the egg. Larvae forced out from the eggs by coverslip pressure are surrounded by 2 cuticles, revealing 2 previous ecdyses. It is thus the third-stage larva that emerges from the hatched egg and not the second-stage larva as previously believed. The cuticle of the first-stage larva is thin, smooth, and remains in the egg, whereas the cuticle of the second-stage larva is thick, striated, and, serving as a cover, loosely ensheaths the released larva. PMID- 8410555 TI - In vitro sensitivity of Plasmodium falciparum to drugs that bind DNA or inhibit its synthesis. AB - The in vitro sensitivity of Plasmodium falciparum to bleomycin, busulfan, camptothecin, CC-1065, cisplatin, daunomycin, distamycin, luzopeptin, mAMSA, mitomycin C, naladixic acid, U71184, VM26, and VP16, or combinations of them was examined. The lethal dose concentration for 50% of parasites for 3 compounds, luzopeptin, CC1065, and U71184, that bind to adenine-thymidine-rich nucleotide sequences, were in the range of 10(-11) M. They were at least 2 orders of magnitude more effective than the other compounds. PMID- 8410556 TI - Infection of immunodeficient mice with a mouse-adapted substrain of the gray strain of Babesia microti. AB - A mouse-adapted substrain was recovered from a severe combined immunodeficient (SCID) mouse inoculated with the Gray strain of Babesia microti. In contrast to the original Gray strain, infection with this substrain (Gray/mo strain) resulted in a high level of parasitemia for a long period in SCID and nude mice, whereas immunocompetent BALB/c mice controlled the infection. The infection profile with the Gray/mo strain was closer to that of human babesiosis than that with the original Gray strain, suggesting usefulness of this mouse model for immunological studies of babesial infection. No antibody activity was revealed in sera of the infected SCID and nude mice. Although strong antibody activity was shown in the sera of the recovered BALB/c mice, no protective activity was demonstrated by the immune serum. Adaptive transfer of thymocytes from the normal BALB/c mouse resulted in SCID and nude mice being able to control the infection, indicating that cell-mediated immunity played a major role in control of infection with B. microti. However, because the reconstituted mice also produced significant antibody titer, antibody remains a possibility for a supportive role in control of the infection. PMID- 8410557 TI - Establishment of the foreign partheonogenetic tick Amblyomma rotundatum (Acari: Ixodidae) in Florida. AB - The parthenogenetic tick Amblyomma rotundatum, a Central and South American species, has become established in southern Florida. The date of introduction is unknown, but it is suspected to be either during the 1930s, when 1 of its natural hosts, the giant or marine toad, Bufo marinus, was introduced to southern Florida as a potential biological control of pest beetles in sugar cane fields, or between 1955 and 1964 when specimens of B. marinus were accidentally or deliberately released in the greater Miami area. Several museum specimens of this toad collected in the Miami area 25 April 1979 had nymphal and adult A. rotundatum attached. Subsequent examination of living giant toads collected at another Miami area site from 1983 through 1985 revealed larval, nymphal, and adult A. rotundatum and confirmed colonization of this tick. Under laboratory conditions, another neotropical amphibian and reptile tick, Amblyomma dissimile, is capable of transmitting Cowdria ruminantium, the causative agent of heartwater, a disease present in the Caribbean area. Therefore, we suggest that A. rotundatum should also be tested for vectorial competence. PMID- 8410558 TI - Occurrence and prevalence of bot flies, Metacuterebra apicalis (Diptera: Cuterebridae), in rodents of cerrado from central Brazil. AB - The occurrence and prevalence of Metacuterebra apicalis (Diptera: Cuterebridae) in natural populations of Oryzomys subflavus, Bolomys lasiurus, and Thalpomys cerradensis (Rodentia: Muridae) from August 1990 to July 1992 in the cerrado, a common savanna-like vegetation type of central Brazil, are reported. An increase in bot infection in the 3 rodent species between October 1991 and July 1992 without correlation to precipitation was detected. The prevalence was lower than in neotropical forest formations. Mean intensity was 1.3 bots (range 1-2) for T. cerradensis and 1 bot for B. lasiurus and O. subflavus. This is the first record of T. cerradensis as host of bot flies. PMID- 8410559 TI - "Microbiological evaluation and monitoring of cleanroom environments" (meeting summary). AB - It was clear that there was little consensus among the panel with regard to the need for an informational chapter in USP on microbial classification of cleanrooms. The articulated positions of the individual panelists remained essentially unchanged throughout the session. What agreement was achieved was in a willingness to continue the dialogue, to await the outcome of the PDA activities cited above with the hope that a mutually acceptable compromise can be realized through that process. PMID- 8410560 TI - In-process protein degradation by exposure to trace amounts of sanitizing agents. PMID- 8410561 TI - Studies in phlebitis. VI: Dilution-induced precipitation of amiodarone HCL. PMID- 8410562 TI - Chemical stability of the cardioprotective agent ICRF-187 in infusion fluids. AB - The chemical stability of the cardioprotective agent ICRF-187 in 0.9% sodium chloride and 5% dextrose infusion fluids has been investigated. The admixtures (concentration: 10 mg/mL and 1 mg/mL) were stored in glass bottles or polyvinyl chloride containers at 4 degrees C in the dark and at room temperature (20-22 degrees C) both protected from light and under normal room fluorescent light conditions in a day-night rhythm. Samples withdrawn immediately after preparation and after 6 hours, 1, 2, 3 and 6 days were analyzed using high performance liquid chromatography with ultraviolet detection. Samples were also inspected for visual changes and tested for changes in pH. Chemical stability of ICRF-187 was also investigated as a function of pH (range 1-12) at room temperature. It is concluded that ICRF-187 is slightly more stable in 5% dextrose than in 0.9% sodium chloride infusion fluids. The stability of the drug is not influenced by normal room fluorescent light nor by the type of container material used. Precipitation occurred in both 5% dextrose and 0.9% sodium chloride with a drug concentration of 10 mg/mL and storage in the refrigerator at 4 degrees C. The chemical stability of ICRF-187 in aqueous solution is mainly a function of pH. At pH 1, no decomposition is detected within 24 hours, at pH 7, 35% decomposition occurs in 21 hours, while at pH 12 it degrades completely within 20.5 hours. PMID- 8410563 TI - Accumulation model for solutes leaching from polymeric containers. AB - An accumulation model for predicting the equilibrium solution concentration of leachables migrating from polymeric containers has been applied to the leaching of several solutes from a blend of a styrene-butadiene-styrene block co-polymer and polypropylene. The model considers three accumulation-limiting mechanisms: total available pool, solute solubility and solute partitioning. Equations relating a solute's solvent-water partition coefficients (Po/w and Ph/w) and its polymer partitioning properties have been developed. With these equations, one can predict leachable accumulation from the container weight, solution volume and the solute's partition coefficients. The maximal accumulation of the leachable in solution will be the lowest value predicted via the three accumulation-limiting mechanisms. PMID- 8410564 TI - The impact of an autoclave cycle on the chemical stability of parenteral products. PMID- 8410565 TI - The generalized form of electrical resistance for a frozen matrix of an aqueous solution. PMID- 8410566 TI - Compliance program for auditing the Chemistry, Manufacturing and Controls section of NDAs and ANDAs. PMID- 8410567 TI - Enhanced stability of two model proteins in an agitated solution environment using poloxamer 407. AB - To enhance the physical stability of two model proteins during solution agitation, we investigated the interaction of the nonionic surfactant poloxamer 407 (Pluronic F-127) with each protein. Vigorous agitation of aqueous solutions of interleukin-2 and urease which contained no poloxamer 407 and were maintained at 4 degrees C resulted in a greater than 50% loss in the biological activity at 12 and 24 hours, respectively. Similar aqueous solutions which were maintained at 4 degrees C and contained either urease or interleukin-2 and poloxamer 407 at a concentration of 10% w/w and 0.5% w/w, respectively lost negligible biological activity when left undisturbed for 96 hours. Moreover, when aqueous solutions of urease and interleukin-2 which contained poloxamer 407 at a concentration of 10% w/w and 0.5% w/w, respectively were maintained at 4 degrees C and subjected to agitation for 96 hours, no significant loss in the biological activity was observed for either protein. In addition, urease was observed to have increased enzymatic activity at early time points regardless of the hydrodynamic solution conditions and poloxamer 407 concentrations evaluated. In contrast, a negligible enhancement in the biological activity of interleukin-2 was observed when aqueous solutions of the protein were exposed to similar hydrodynamic conditions employed for urease solutions, but different poloxamer concentrations (0% w/w vs. 0.5% w/w). Results of molar ellipticity, [theta], versus wavelength, lambda, profiles using CD spectropolarimetry on individual aqueous solutions of both proteins containing 2% w/w poloxamer 407 were in close agreement to spectrum obtained with each protein in pH = 7 phosphate buffer (PB).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410568 TI - The influence of disabling condition visibility on family functioning. AB - Examined whether the level of family functioning and the components contributing to adaptive family functioning differed in families of visibly handicapped children (cerebral palsy) when compared to families of nonvisibly disabled children (diabetes). Other factors included effect of disability severity on family functioning, comparison of families of disabled children to families of able-bodied children, and comparison of mothers' and fathers' perceptions of family functioning. The sample comprised 139 two-parent families of children with cerebral palsy (n = 48), diabetes (n = 46), and able-bodied children (n = 45) (all 5-11 years old). Results showed that neither visibility nor severity of disability differentially impacted family functioning. Furthermore, families of the disabled children exhibited high levels of family functioning which were similar to control families. Differences were not found between the ways mothers and fathers perceived family functioning. PMID- 8410569 TI - Associations among teen-parent relationships, metabolic control, and adjustment to diabetes in adolescents. AB - Extended previous studies of adolescents with insulin-dependent diabetes mellitus (IDDM) that have implicated family conflict as a correlate of poor adaptation to the disease and inadequate diabetic control. Families of 115 adolescents with IDDM completed the Parent-Adolescent Relationship Questionnaire (PARQ) and the Teen Adjustment to Diabetes Scale (TADS) and recent glycohemoglobin levels were retrieved from medical records for 82 patients. PARQ scores of families of adolescents with IDDM were similar to those of a healthy normative group. Multiple regression analysis showed that a PARQ scale measuring family communication and conflict resolution skills was a strong predictor of the IDDM outcome variables, suggesting that these families could derive health and behavioral benefits from an intervention that improves parent-adolescent communication skills. Controlled trials of such interventions are needed. PMID- 8410570 TI - Development of the Child Attitude Toward Illness Scale. AB - Described the development of the Child Attitude Toward Illness Scale (CATIS), a short self-report instrument designed to provide a systematic assessment of how favorably or unfavorably children feel about having a chronic physical condition. Subjects were children (N = 269), ages 8-12 years, who had either epilepsy (n = 136) or asthma (n = 133). Results of confirmatory factor analysis and predicted relationships with scores on the Child Behavior Checklist and the Piers-Harris Children's Self-Concept Scale provided support for construct validity of the scale. Good internal consistency and test-retest reliability were also found. PMID- 8410571 TI - Predicting children's presurgical anxiety and subsequent behavior changes. AB - Addressed, in two studies, issues of children's medical anxiety having implications for efficient psychological preparation. Study 1 assessed behavioral distress and physiological arousal of 4- to 10-year-old, unprepared, nonsedated children (N = 50) as they separated from parents, waited in the operating room, and were given general anesthesia for minor surgery. Anxiety intensity varied widely among children and was most intense at mask presentation. Mother's prediction of uncooperative behavior and a history of prior surgery were the best predictors. Study 2 examined anxiety of 37 children prior to surgery and behavior changes 2 weeks later. At follow-up, minor or transient problems were common; 4 children developed significant problems. Elevated presurgical anxiety predicted later problems, but only among children hospitalized after surgery. Findings suggest that presurgical anxiety and later behavior problems are partially predictable. PMID- 8410572 TI - Society of Pediatric Psychology Task Force Report: pediatric psychology and injury control. AB - Injuries are the major causes of death for children. Pediatric psychology offers significant contributions to the multidisciplinary efforts necessary to prevent injuries and reduce harm to children. This Task Force Report reviews epidemiological data, characteristics of children's injuries, passive and active interventions for reducing injuries, and research, policy, and evaluation issues for individual and community injury control efforts. Directions for future pediatric psychology efforts are identified and placed in a context of collaborative efforts required to advance the control of children's injuries. PMID- 8410573 TI - 19th annual Frank Costenbader Lecture--the origins of congenital esotropia. AB - Congenital esotropia develops in the first 4 months of life in an infant who lacks the inborn mechanism for motor fusion. It manifests as an esotropia which is not eliminated by correction of hyperopia and occurs in an otherwise neurologically normal infant. The earliest practical time for surgery is 4 months of age. The eye is anatomically suited for surgery at this age and also, this is the earliest age that congenital esotropia can be diagnosed with confidence. The best attainable result of treatment of congenital esotropia is subnormal binocular vision. This result is more likely to be attained if infants are aligned by 18 months of age. Satisfactory alignment is produced in 80% to 85% of infants in one procedure with an appropriate bimedial rectus recession. An array of motor defects including DVD, latent nystagmus, oblique dysfunction, and A- and V-pattern appear at varying times after successful alignment. These associated findings are commonly found with, but are not unique to, congenital esotropia. The onset and clinical picture of congenital esotropia is satisfactorily explained by a theory first suggested by Worth that the strabismus is caused by an inborn defect in the motor fusion mechanism and aggravated by esotropital factors as suggested by Chavasse. In contrast to congenital esotropia, all other strabismus can be thought of as occurring on a secondary basis in a person with the inborn capacity for motor fusion, but who failed to maintain it because of conatal insurmountable strabismus (congenital third nerve palsy), who lost it because of acquired (postnatal) strabismus, who uses a strategy such as head posture alteration to retain fusion under favorable circumstances (Duane syndrome), who has intermittent strabismus with part-time suppression (X(T)), or who is maintaining alignment with nonsurgical means (refractive esotropia). For the future, I believe that advances in the management of congenital esotropia will depend on a better understanding of etiology leading to design and use of innovative nonsurgical techniques to discourage convergence and stimulate bifoveal fusion. PMID- 8410574 TI - Binocularity in accommodative esotropia. AB - One hundred twenty-seven medical records of patients with accommodative esotropia met our inclusion criteria and were analyzed. All included patients were within 10 prism diopters of orthophoria and had stereopsis and other binocular sensory test results recorded at their latest visit. Bifixation, defined as stereopsis of 50 arc seconds or better, was present in 31 patients (24%) with an average follow up of 89 months. Monofixation (peripheral fusion) was present in the remaining 96 patients (76%) with an average follow up of 84 months. Patients with bifixation were less likely to have presented with constant esotropia (19% vs 39% [P = .04]) and were more likely to be aligned within 8 delta of orthophoria in their first glasses (84% vs 21% [P < .0001]). No patient with bifixation had constant esotropia longer than 4 months. In addition, patients with bifixation were less likely to have worn bifocals (39% vs 59% [P = .09]), or undergone esotropia surgery (23% vs 62% [P < .0001]). These data suggest that maintenance of bifixation is possible in accommodative esotropia if the eyes are straightened before or shortly after the esodeviation becomes constant. With this early therapy, amblyopia and deterioration of ocular alignment are also less likely. PMID- 8410575 TI - A new VEP system for studying binocular single vision in human infants. AB - Visual evoked potentials (VEPs), that provide unequivocal objective evidence of cortical binocularity have been recorded from adults and young infants using a new VEP system developed for this purpose. The system uses alternating field stereoscopy (AFS) to present separate visual stimuli to each eye. With this system, the binocular image pairs to the right and left eyes alternate at a high rate on a single video monitor. The subject wears spectacles incorporating light scattering liquid crystal lenses which alternate electronically between opaque and clear modes in synchrony with the video monitor. To detect cortical binocularity, the system analyzes VEP activity mathematically and identifies significant responses at test frequencies reflecting binocular cortical interactions exclusively. Three types of binocular stimuli were presented: (1) dynamic random dot correlograms (correlograms); (2) dynamic random dot stereograms (stereograms); and (3) dichoptic checkerboard stimuli. The correlograms are generated when moving random dot patterns presented to each eye alternate between two phases, correlated and anticorrelated. With the stereograms, portions of random dot patterns presented to each eye are shifted horizontally relative to each other at a fixed rate, alternately producing crossed and uncrossed binocular disparities. Subjectively, these patterns appear to shift in depth. Dichoptic checkerboard stimuli are regular checkerboard patterns which reverse at different rates (frequencies) for each eye. Binocular VEPs are generated due to cortical interactions at the difference (beat) frequency. Using this VEP system, we have recorded binocular VEPs from 10 normal adults and more than 40 infant subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410576 TI - Compliance with safety glasses wear in monocular children. AB - Many ophthalmologists prescribe polycarbonate "safety glasses" to protect the remaining eye of monocular patients. Others do not routinely do so when the remaining eye is emmetropic since they feel compliance is poor. To evaluate compliance with safety glasses wear in these children, we reviewed the charts of all children (< 21 years old) treated with enucleation and followed at the University of Iowa by the Pediatric Ophthalmology Service between 1962 and 1991. Safety glasses were defined as polycarbonate lenses in a frame suitable for protective wear with spherical equivalent < or = -0.75 or < or = +2.00 diopters. Fifty-six patients were followed after enucleation. Forty-six of these patients met the inclusion criteria and 33 patients were available for follow up. Eighty five percent of patients wear safety glasses > or = 50% of their waking hours; 61% wear them > or = 80% of the time; 33% wear them 100% of their waking hours. Twenty-one of the 33 patients participate in sports. One hundred percent of the female participants and 80% to 93% of the male participants wear safety glasses, goggles, or face shields during sports. Fifteen of the 33 respondents reported at least one potentially serious accident in which the safety glasses had protected the good eye. Compliance with safety glasses wear appears to be good in this population and may prevent injury to the remaining eye. PMID- 8410577 TI - Pediatric aphakic contact lens wear: rates of successful wear. AB - Two hundred thirty-eight consecutive patients with pediatric cataracts were fitted with a variety of aphakic contact lenses after cataract extraction. Thirty nine patients did not return for follow up. Of the remaining 199 patients, 40 discontinued contact lens wear. None of the 78 patients with unilateral or bilateral acquired cataracts discontinued contact lens wear due to problems wearing their lenses, although nine discontinued contact lens wear due to poor vision or difficulty maintaining amblyopia treatment. Twenty-two of 84 patients with unilateral congenital cataracts discontinued lens wear, six directly due to difficulties wearing lenses. Sixteen had poor vision in their aphakic eye and inability to maintain patching for amblyopia. Eight of 37 patients with bilateral congenital cataracts discontinued lens wear because of problems wearing their lenses, and one other discontinued lens wear because of poor vision. In summary, 26 of 40 patients (out of a total of 199) that discontinued aphakic contact lens wear did so because of poor vision, while only 14 did so because of difficulties wearing the contact lenses. Eleven of these 14 patients were able to wear aphakic spectacles in lieu of contact lenses. This study shows that most pediatric patients with cataracts fail at treatment because of problems related to treatment of amblyopia, and not problems related to the fitting and wearing of contact lenses. PMID- 8410578 TI - The effect of grating orientation on resolution acuity in patients with nystagmus. AB - The acuity card procedure is an effective method for the assessment of grating acuity in preverbal and nonverbal patients. However, the effect of nystagmus on this acuity assessment has not been well-described. We studied the effect of grating orientation on the measurement of resolution acuity in a group of nystagmus patients, a group of patients with albinism and nystagmus, and a normal control group. The resolution acuity was significantly (P < or = .01) better with horizontal grating orientation in both the nystagmus and the albino groups, but grating orientation did not affect resolution acuity in the control group. This study suggests that grating orientation must be taken into account when using the acuity card procedure in patients with nystagmus. PMID- 8410579 TI - Epstein-Barr virus infection associated with encephalitis and optic neuritis. PMID- 8410580 TI - Retinal dystrophy in the cardiofaciocutaneous syndrome. PMID- 8410581 TI - Atypical iris colobomata and Pfeiffer syndrome. PMID- 8410582 TI - Lyme amaurosis in a child. PMID- 8410583 TI - Congenital dacryocystitis and retrobulbar abscess. PMID- 8410584 TI - Adolescent fitness is a challenge. PMID- 8410585 TI - Pap smear screening for adolescents: rationale, technique, and follow-up. AB - Cervical cytologic abnormalities are increasingly prevalent in teenagers. Adolescents are at greater risk for cervical neoplasia than adult women because of the biologic changes occurring in the cervix during puberty, the prevalence of human papillomavirus, and the behavioral risk factors in this age group. Two behavioral risk factors are early age at first coitus and multiple sex partners. Human papillomavirus is widely believed to be the etiologic agent associated with the spectrum of cervical neoplasias. Papanicolaou smear technique, reporting, and management of abnormal smears in teens is explained. PMID- 8410586 TI - Preventing day-care--related illnesses. AB - The number of children enrolled in out-of-home day care has increased dramatically. One of the concerns expressed about the use of day care is the contribution of group settings to childhood morbidity. Children who are placed in groups of other children experience more illnesses than their peers who remain at home. However, disease transmission can be minimized by infection control behaviors. Unfortunately, day-care staff and parents are often inadequately prepared in such behaviors. Consequently, by becoming involved in day care, nurses can play a role in reducing the number of day-care--related illnesses. PMID- 8410587 TI - Test-retest and interobserver reliability of the Early Language Milestone Scale, second edition. AB - Test-retest and interobserver reliability are reported for the Early Language Milestone Scale, second edition, a language assessment tool for infants and children from birth to 36 months of age. Reliability data are presented for two scoring methods: the pass-fail method that yields a global rating of "pass" or "fail," and the point-scoring method that yields percentile scores. Both scoring methods showed good reliability. The clinical applicability of each scoring system is discussed. PMID- 8410588 TI - Parental knowledge and practices of skin cancer prevention: a pilot study. AB - Skin cancer is a growing health threat in the United States; effective prevention logically begins with sun avoidance practices initiated early in childhood. However, little is known about the current level of skin cancer knowledge or skin cancer prevention practices among parents. A pilot survey of these levels was conducted among parents attending a single pediatric office in a nationally affiliated health maintenance organization. This article reports the findings of that survey and identifies areas of knowledge and practice deficit. Implications and interventions are discussed. PMID- 8410589 TI - The child with a limp. PMID- 8410590 TI - School phobia/school avoidance. PMID- 8410591 TI - Heart murmur. PMID- 8410592 TI - Nurses achieve victory on prescriptive authority at DEA. PMID- 8410593 TI - Shaken baby syndrome. PMID- 8410594 TI - Evaluating a private practice for employment. PMID- 8410595 TI - NAPNAP position statement. National Association of Pediatric Nurse Associates and Practitioners. AB - This position statement outlines what NAPNAP believes to be the "bottom line" elements of child health care that must be incorporated into any health care reform proposal. NAPNAP's focus on the optimal physical, mental, emotional, and social health of the nation's children encourages them to grow and develop to their fullest potential. PMID- 8410596 TI - 1992 NAPNAP membership survey. Part I: Member characteristics, issues, and opinions. National Association of Pediatric Nurse Associates and Practitioners. AB - Significant changes have occurred in the 4 years since the last NAPNAP membership survey was conducted. Membership in NAPNAP has shown strong, sustained growth; the role of nurses in advanced practice has expanded greatly, and an increasing possibility exists for national health care reform. In response to these changes and as a part of its long-range strategic planning, the NAPNAP Executive Board, in 1991, authorized an ongoing plan to conduct a membership survey every 4 years. The 1992 membership survey was mailed to a random sample of 800 NAPNAP members. Sixty-five percent were returned. The survey collected information on (a) demographic characteristics of NAPNAP members, (b) practice characteristics of NAPNAP members, and (c) opinions of NAPNAP members related to health care issues. This article presents summary data on demographic characteristics, issues, and opinions of NAPNAP members. PMID- 8410597 TI - Efficient and unbiased estimation of volume and area of tissue components and cell number in gingival biopsies. AB - Recently, the methodology for volume and cell number estimation from sectioned tissue has undergone rapid progress. To date there are no indications in the literature that investigators in periodontal research are aware of this progress, published mainly in the disciplines of stereology and morphometry. This is unfortunate, as these developments could be profitably exploited in periodontal pathology. In this review we discuss the dangers of some currently used quantitative methods and indicate how simple unbiased methods together with an appropriate systematic sampling scheme lead to an efficient estimation of cell number and volume of tissue or tissue components. PMID- 8410598 TI - Transmission of Actinobacillus actinomycetemcomitans in families of adult periodontitis patients. AB - At present Actinobacillus actinomycetemcomitans is regarded as an important microorganism in the etiology of some forms of periodontitis. The purpose of the present investigation was to study the number of Restriction Endonuclease Analysis (REA)-types present in the oral cavity of A. actinomycetemcomitans positive subjects and to study the possibility of transmission of A. actinomycetemcomitans within families of adult periodontitis patients. DNA of A. actinomycetemcomitans isolates was digested with a combination of the restriction endonucleases PstI and BamHI, after which the DNA fragments were separated by agarose gel-electrophoresis. To study the number of REA-types, multiple A. actinomycetemcomitans isolates obtained from 8 different sites in the oral cavity of five subjects were typed. The results showed that in most cases only one REA type is present. In the 13 families investigated in 4 of the 26 children (15%) and in 1 of the 13 spouses (8%) of the adult periodontitis patients an indistinguishable REA-type was found within the families. This suggests that also in the case of adult periodontitis transmission of A. actinomycetemcomitans is possible, but does not seem to occur easily. PMID- 8410599 TI - Effect of anaerobiosis and sulfide on killing of bacteria by polymorphonuclear leukocytes. AB - Anaerobic microorganisms in periodontal pockets produce toxic amounts of hydrogen sulfide. The capacity of polymorphonuclear leukocytes to kill a capsulated and a non-capsulated variant of a group B streptococcal strain was studied in presence and absence of sulfide. The killing was equally efficient under aerobic and anaerobic conditions. However, in presence of sulfide the killing of the capsulated variant of the strain was significantly inhibited. Since this strain required higher serum concentrations to be killed by the polymorphonuclear leukocytes, it suggested that sulfide interfered with the opsonization of the bacteria. The capacity of sulfide to split the disulfide bonds of complement factor 3 and immunoglobulin G, deposited on the bacterial surface, was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was no detectable effect of 2 mM sulfide on immunoglobulin G. However, sulfide released from opsonized bacteria the beta-chain of C3b C3bi, and the C-terminal part of the alpha-chain of C3bi. This region of the alpha-chain of C3bi has been suggested to bind to the complement receptor 3 of polymorphonuclear leukocytes. The beta-chain of C3b/C3bi may augment the binding of opsonized bacteria to the complement receptors of polymorphonuclear leukocytes. The formation of sulfide by the microflora of the periodontal pockets may provide conditions for the bacteria to escape important parts of the host immune system. PMID- 8410600 TI - Expression of TIMP-1, TIMP-2 and collagenase mRNA in periodontitis-affected human gingival tissue. AB - Collagenolysis in periodontitis is thought to be modulated by the expression of three genes, collagenase, tissue inhibitors of metalloproteinases-1 and -2 (TIMP 1 and -2). We assessed the possible difference in TIMP-1, TIMP-2 and collagenase mRNA levels between gingival samples from patients with periodontitis and those from healthy subjects by reverse transcription-polymerase chain reaction (RT PCR). This technique allows detection of transcripts from a very small sample quantity. The experiments showed that levels of TIMP-1 and collagenase transcripts relative to beta-actin are significantly higher in the diseased group than in healthy controls (8.11 +/- 0.83 versus 1.38 +/- 0.28% for TIMP-1 and 0.50 +/- 0.10 versus 0.0075 +/- 0.0024% for collagenase, respectively). The difference in TIMP-2 between the two groups (2.91 +/- 0.46 versus 1.84 +/- 0.87%) did not differ. Therefore, the host would have responded to the increase in collagenase level by preferentially producing TIMP-1 against tissue destruction. The differential gene expression of TIMP-1 and TIMP-2 in our study may account for a distinct genetic regulation of TIMP-1 and -2 in vivo. PMID- 8410601 TI - Differential expression of CR3, Fc epsilon RII and Fc gamma RIII on polymorphonuclear leukocytes in gingival crevicular fluid. AB - Polymorphonuclear leukocytes (PMNLs) are the most numerous cell population among the cellular infiltrates in gingival crevicular fluid (GCF) and play important roles in the host-defensive system in the gingival crevices. We determined the percentage of neutrophils, eosinophils and basophils in total PMNLs by light microscopic observation using Randolph-methylene blue staining, then assessed flow cytometric differences in the expression of CR3, Fc gamma RIII, Fc epsilon RII, LFA-1 alpha, and LFA-1 beta on PMNL in GCF and peripheral blood (PB) from 21 patients with adult periodontitis (AP) and 13 healthy donors. Percentages of basophils and eosinophils were higher in GCF than in PB. In both AP patients and healthy subjects, expression of CR3 and Fc epsilon RII was higher while Fc gamma RIII was lower in GCF than in PB. The statistical analysis showed that the expressions of Fc gamma RIII and Fc epsilon RII on GCF PMNLs were lower in AP patients than in healthy subjects. Expressions of LFA-1 alpha and beta on GCF were similar to those on PB PMNLs. PB PMNLs stimulated in vitro with Porphyromonas gingivalis culture supernatant and fMLP displayed an expression pattern of CR3, Fc gamma RIII and Fc epsilon RII on GCF PMNLs. However, C5a and IL-1 failed to induce changes in Fc gamma RIII and Fc epsilon RII. The results indicate that GCF neutrophils are activated, present enhanced adhesion and a decreased IgG-binding ability which would reflect that they are at the terminal stage of activation, and that GCF contains a larger eosinophil fraction than in PB. Moreover, these GCF eosinophils appear to be activated. PMID- 8410602 TI - Disposition of nifedipine in plasma and gingival crevicular fluid in relation to drug-induced gingival overgrowth. AB - The present study investigates the relationship between the pharmacokinetic variables of nifedipine with the incidence and severity of gingival overgrowth in 9 adult male patients medicated with the drug for at least 6 months. Five of the patients had experienced significant gingival changes and were thus designated "responders". The remaining four patients exhibited no gingival overgrowth, and thus acted as a control. A baseline periodontal examination (plaque scores, bleeding index and gingival overgrowth assessment) was carried out on each patient, and confined to the upper and lower anterior teeth. Serial blood and gingival crevicular fluid samples were collected over an eight-hour investigation period. Samples were analyzed for nifedipine by gas chromatography. No significant difference (p > 0.05) was seen between responders and non-responders with regard to drug therapy, periodontal parameters or plasma pharmacokinetics of nifedipine. Nifedipine was detected in the gingival crevicular fluid of seven subjects (all responders, and two non-responders). The peak concentration of nifedipine in crevicular fluid was 15-90 fold greater than levels observed in plasma. PMID- 8410603 TI - Inhibition of epithelial cell matrix metalloproteinases by tetracyclines. AB - The effects of tetracyclines on periodontal epithelial cells were investigated by culturing cells from porcine rests of Malassez in the presence of oxytetracycline, doxycycline or one of two analogues of tetracycline bearing no antimicrobial activity. Matrix metalloproteinase activity produced by the epithelial cells was assayed by quantitation of radioactive gelatin degradation and by gelatin enzymography. The results show that all tested tetracyclines exerted a direct dose-dependent inhibitory effect on epithelial cell gelatinases. Furthermore, epithelial cells cultured with doxycycline, oxytetracycline and de dimethylaminotetracycline in concentrations ranging from 1 to 50 micrograms/ml showed a marked reduction in secreted gelatinase activity when grown in alpha minimum essential medium in the absence of fetal calf serum. Viability of cells following this treatment, measured as lactate dehydrogenase activity released to the cell media, was not affected by the presence of any of these drugs at the concentrations used. Scanning electron microscopy revealed striking morphologic changes of the cells following treatment with tetracyclines in the absence of serum which include rounding, decreased intracellular contacts and increased intercellular spaces. No such effects were seen in cells cultured in the presence of serum. These results provide evidence that periodontal epithelial cells produce matrix metalloproteinases whose activities are inhibited by tetracyclines and their non-antimicrobial analogues at concentrations present in gingival crevicular fluid following tetracycline therapy. When used as adjuncts in periodontal therapy, tetracyclines may therefore inhibit epithelial cell mediated degradation of basement membrane and subepithelial connective tissue. PMID- 8410604 TI - The double lateral bridging flap for coverage of denuded root surface: longitudinal study and clinical evaluation after 5 to 8 years. AB - Longitudinal results of a 5 to 8 year follow-up study of the double lateral bridging flap for coverage of gingival recessions (GR) are presented. On a total of 75 GR in 18 patients (31 surgical procedures) the majority of the teeth (58%) showed a root denudation reduction of 75% or more. Total coverage was observed on 18 teeth (24%). Examinations did not show a high correlation between the extent of recession and bone dehiscence after surgery, between the degree of root coverage and the initial width of keratinized gingiva, or between the GR before and after surgery. We conclude that neither the quantity of gingival recession nor the quality of the supporting tissues were a prerequisite for the success of this surgical technique. PMID- 8410605 TI - Comparative SEM study on the effect of acid etching with tetracycline HCl or citric acid on instrumented periodontally-involved human root surfaces. AB - This study compared the surface characteristics of periodontally diseased single rooted human teeth extracted after treatment with either tetracycline HCl or citric acid solutions. The study group was comprised of 30 teeth from 22 patients with advanced periodontal disease extracted before the start of periodontal therapy. Diseased surfaces were identified, outlined, and root planed by hand curet or finishing bur. The teeth were sectioned and solutions of tetracycline HCl or citric acid (pH 1) were applied to the surfaces with cotton pellets for 5 minutes. Extracted teeth were processed and root surface samples then examined by scanning electron microscope. The surfaces of both acid-treated sets of specimens differed considerably from specimens treated with root planing alone, regardless of root planing method. Acid-treated specimens exhibited dentinal tubules exposed by the removal of the smear layer, surfaces devoid of the debris normally present in root planed-only specimens, and the dense network of collagen fibers that make up the dentin structure. Although differences were seen in surface depressions and fiber-like structures among some specimens, the tetracycline HCl and citric acid solutions produced comparable morphologic characteristics. PMID- 8410606 TI - Comparative analysis between a modified ultrasonic tip and hand instruments on clinical parameters of periodontal disease. AB - The goal of this study was to determine whether an ultrasonic scaler with a modified tip is as effective as a curet in providing supportive periodontal treatment for patients, based on clinical parameters of periodontal disease. Nine patients with 10 sites exhibiting probing pocket depth > or = 3 mm were treated at 0, 90, and 180 days in a single-blind, split-mouth design for supportive periodontal treatment with either Gracey curets (GC) or an ultrasonic scaler with a modified tip (MU). Clinical parameters included plaque index, gingival index, bleeding on probing, darkfield microscopy, and elastase presence. Probing pocket depths and attachment levels were measured using an electronic probe. Measurements of clinical parameters were taken at 0, 14, 45, 90, 135, and 180 days. The results showed that treatment with MU was as effective as treatment with GC in all clinical parameters measured. Both treatment modalities were effective in reducing the elastase levels. Instrumentation time was significantly reduced with the MU (3.9 minutes vs. 5.9 minutes, P < 0.05). The MU instrument effectively reduced the microbial environment in a significantly shorter time as compared to GC. PMID- 8410607 TI - Survival characteristics of periodontally-involved teeth: a 40-year study. AB - Four hundred and fifty-five teeth that were judged clinically to have a questionable prognosis were observed over a 40-year span in 166 patients in a private practice. Therapy initially was debridement with oral hygiene instruction, followed by flap curettage procedures and preventive maintenance recall therapy. An assessment of compliance based on oral hygiene and recall attendance was also performed. A total of 55 (12%) teeth were lost with an average survival rate of 8.8 years prior to extraction. No teeth were lost during the first 2 years of observation. The majority of extracted teeth were maxillary second molars (38.2%), followed by maxillary first molars (25.5%), and mandibular second molars (16.4%). Tooth loss patterns appeared to be bilaterally symmetrical, with 51% of teeth lost in the right quadrants and 49% lost in the left quadrants. Indications for extraction were primarily periodontal abscesses. Teeth that were considered to have a very questionable prognosis have been retained for many years after therapy, supportive treatment, and patient compliance with recall attendance. PMID- 8410608 TI - Epithelial cell kinetics with atelocollagen membranes: a study in rats. AB - A recent development in guided tissue regeneration procedures is the use of resorbable membranes, which eliminate the need for subsequent surgical removal. In this study we performed flap surgery in rats with (experimental) or without (control) implantation of one of the newer materials, atelocollagen. We observed the gingival epithelial cell kinetics using 3H-thymidine and examined the extent of gingival epithelium migration. Histological observations at day 1 on the experimental side demonstrated regenerated epithelium apposed to the collagen membrane with an intervening layer of necrotic tissues and/or fibrinous exudate. There was no observable proliferation of regenerated epithelium toward the root apex. On day 14, the regenerated epithelium migrated apically along the treated root surface in the control group. By contrast, on day 14 in the experimental group, the regenerated epithelium contacted the root surface at the cemento enamel junction (CEJ). Apical to the CEJ, there was new cementum formation with inserting connective tissue fibers. Autoradiographs from day 1 experimental sides demonstrated labeled cells in the basal cell layers from oral, crevicular, and junctional epithelium. From day 1 to day 5, labeling indices of oral epithelium and regenerating crevicular epithelium on experimental sides were lower than on control sides. These histological and autoradiographic findings suggest that atelocollagen membrane inhibits apical migration of regenerating epithelium and accelerates connective tissue reattachment in part by inhibiting the mitotic function of basal epithelial cells in early stages of wound healing. PMID- 8410609 TI - The rate of periodontal attachment loss in subjects with established periodontitis. AB - A stepwise approach to determine attachment level changes was utilized to assess the nature of progression of periodontal disease. Following initial screening, 51 subjects with established periodontitis were monitored quarterly for 9 more months. Probing depth (PD) and relative attachment level (RAL) were recorded using an automated, pressure sensitive probe system. To establish intra-examiner error, repeated measurements were performed for all sites at the final visit. An overall standard deviation (SD) for RAL repeated measurements was initially calculated (0.76 mm) using all 6,935 double measurements. Sites were sorted by factors which contribute to the error of attachment level measurements; i.e., pocket depth (shallow, moderate, deep), tooth type (molar, non-molar) and location (buccal, lingual). Data were sorted by the above 12 groups, and SD for repeated measurements was calculated separately for them. The ratio between these SD and the overall SD served as the corrective factor. Each patient's initial threshold (2 SD) was multiplied by these corrective factors thus resulting in 12 thresholds for each subject. Next, linear, exponential and logarithmic regression models were tested for each site, and the regression model showing the highest R value was chosen for that site. AL changes were tested against the patient's threshold for that site. Sites with attachment loss exceeding the threshold were deemed active. Five hundred eighty-one sites (8.3%) exhibited attachment loss exceeding the various thresholds. Of these, linear progression occurred in 195, logarithmic in 224, and exponential in 162 sites. Individual patient's attachment loss ranged from 0.6 to 19.4% of all sites.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410610 TI - The effect of chlorhexidine irrigation on tensile wound strength. AB - Chlorhexidine in an alcohol vehicle with flavoring agents has been used as a mouthrinse to reduce plaque accumulation in periodontal surgery patients. The purpose of this study was to evaluate the effects of a chlorhexidine-containing mouthrinse on the early tensile wound strength of healing surgical wounds in the rat. Standardized transdermal incisions were made on each lateral abdominal wall of 40 Sprague-Dawley rats. Wounds were irrigated with 10 ml of 0.12% chlorhexidine or 10 ml of normal saline prior to closure. Animals were sacrificed at 48 hours and 96 hours, and the wound area was excised by a standardized protocol. Wound strength was measured using constant speed tensiometry to determine the tensile strength of the healing incision. Results revealed a significantly reduced tensile wound strength at 48 hours for the chlorhexidine treated group (127 +/- 18.5 gm) compared to the saline irrigation group (150 +/- 32.3 gm) (P < 0.001). However, by 96 hours a significantly increased tensile wound strength was demonstrated by the chlorhexidine treated group (202.1 +/- 21.7 gm) compared to the saline irrigation group (183.2 +/- 37.3 gm) (P < 0.05). These data suggest that chlorhexidine-containing mouthrinse irrigation of wounds produced a reduced early tensile wound strength, but ultimately resulted in shorter healing time. PMID- 8410611 TI - Long-term periodontal care: a comparative retrospective survey. AB - The maintenance of 172 patients who were treated and then attended a periodontal practice in Sydney, Australia for periods of at least 10 years has been surveyed. The results, expressed as loss of teeth over the maintenance period, have been compared with a 1978 study. Results of the two surveys have been compared for total tooth loss over the maintenance period, loss of individual tooth types, loss of teeth with furca lesions, and tooth loss relative to surgical experience. The comparative analysis is limited by the difference in total number of patients and by the duration of the studies. The proportionate division of patients into three groups according to tooth retention (well maintained, downhill, and extreme downhill) was statistically similar in both surveys. There were very few statistically valid differences in numbers of individual teeth lost between the two studies. Despite these comparable end results of long-term maintenance, the surgical experience was quite different, with the patients in our practice undergoing much more surgical treatment than those in the earlier report. The results appear to support the hypothesis that long-term maintenance is attainable for most periodontal patients, and is consistent with a variety of treatment approaches. PMID- 8410612 TI - Surgical treatment of intrabony periodontal defects using expanded polytetrafluoroethylene barrier membranes: influence of defect configuration on healing response. AB - Twenty-six proximal, intrabony periodontal defects with probing depths equal to or exceeding 6 mm in 23 patients were treated with gingival flap surgery supported by an expanded polytetrafluoroethylene barrier membrane. The material included 1, 2, and 3-wall defects with crestal involvement relative to the root circumference ranging between 90 degrees and 270 degrees. Healing was evaluated clinically 12 months after surgery. Deep defects exhibited greater probing depth reduction and attachment gain than shallower defects. Probing depth reduction, attachment gain, and bone fill were positively correlated to the depth of the 3 wall intrabony component of the defect. The extent of crestal involvement, and wall form in the fundus of the defect did not appear to influence the healing response. The treatment also affected the proximal surface of the defect-adjacent tooth, which showed some reduction in bone level as well as gingival recession. Thus, the overall healing pattern following barrier membrane-supported flap surgery appears similar to that generally observed for conventional reconstructive flap surgery in intrabony periodontal defects. PMID- 8410613 TI - Recurrent compound naevus of gingiva. AB - A case is described of a compound naevus of the gingiva. It was unusual in that it showed a number of recurrences and these appeared to be related to the hormonal status of the patient. PMID- 8410614 TI - Periodontal therapy: prospects for the future. AB - Prior to the 1950s, periodontitis was treated mostly by tooth exfoliation or extraction, and that is still the predominant treatment for most of the world's populations today. Debridement of the root surface by scaling and root planning came into relatively common use in the first half of the present century and has become the central feature held in common by all currently-used forms of periodontal therapy. Until the 1980s, the most commonly-used treatment consisted of scaling and root planing, followed by resective surgery aimed at achieving zero pocket depth. During the 1980s, data were obtained demonstrating that the thoroughness of root debridement and subgingival infection control, not the presence or absence or periodontal pockets, is the major determinant of successful periodontal therapy, and non-surgical therapy became a commonly-used treatment. Neither resective surgery nor non-surgical therapy results in significant regeneration of periodontal attachment. With the realization that periodontitis is an infectious process, the use of antibiotics and other anti infective agents came into common use as adjuncts to other standard therapies. An understanding of the pathways by which the soft and calcified tissues of the periodontium are destroyed has led to the likelihood of widespread future use of the non-steroidal, anti-inflammatory family of drugs to suppress alveolar bone destruction by blocking prostaglandin production, and to the use of chemically modified tetracyclines that chelate divalent cations and thereby block tissue destruction by the metalloproteinases. Recent data clearly show that regeneration of the previously-destroyed periodontal attachment tissues is biologically possible, and regeneration has become the goal of therapy for the 1990s. Use of osteoconductive and osteoinductive graft materials can, under favorable conditions, induce roughly 60% to 70% regeneration of bone lesion height or volume with concomitant improvement in the clinical conditions. Regeneration by grafting may be further enhanced by use of barrier membranes that exclude gingival fibroblasts and epithelium from the healing site. Still further enhancement seems to be possible by local application of various growth factors, although studies in this important area are now only in their infancy. The future of periodontal therapy is exceedingly bright.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410615 TI - Effect of therapy on periodontal infections. AB - Periodontal disease progression requires the simultaneous presence of high numbers of pathogens, low numbers of compatible or beneficial species, a conductive local environment, and a susceptible host. Effective therapy acts by altering one or more of these factors. Data from an ongoing study were used to examine the biological basis of treatment success or failure. Seventeen subjects showing disease progression were treated by Widman flap surgery at deep sites, scaling at shallow sites, and 1 of 4 randomly-assigned, systemically-administered adjunctive agents including amoxicillin/clavulanate potassium (Au) (n = 3), ibuprofen (n = 3), tetracycline (n = 9), or a placebo (n = 2). Clinical measurements and microbiological samples (enumerated using DNA probes) taken from the mesial aspect of each tooth pre-treatment and 12 months post-treatment were compared and 418 pre- and 418 post-therapy plaque samples were enumerated. Overall, the 4 treatments resulted in pocket depth reduction and "gain" in attachment. After therapy, the percentage of sites colonized by Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, and Bacteroides forsythus was decreased and counts > 10(6) were less frequent. Large attachment level gains were accompanied by major decreases in these species and were more frequent in subjects receiving antibiotics. A small number of sites in each treatment group became deeper and/or lost attachment. More than half of these sites were detected in 2 subjects who were older (65 vs. 44), had higher serum antibody to Actinobacillus actinomycetemcomitans serotype a (506 vs. 125 ELISA units), A. actinomycetemcomitans serotype b (518 vs. 130), and Campylobacter rectus (39 vs. 18).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410616 TI - Current status of systemic antibiotic usage in destructive periodontal disease. AB - The recognition that periodontal diseases are primarily caused by specific microorganisms has led researchers to explore the possibility that antibiotics may enhance the effect of mechanical debridement procedures such as scaling and surgery. For some selected periodontal diseases, this has proven to be true. This paper will review systemically-administered antibiotics and the clinical studies and case reports supporting their use. In periodontal therapy, the tetracyclines are the most commonly-used antibiotics in the United States. Tetracycline hydrochloride, minocycline, and doxycycline have been shown to inhibit in vitro most putative periodontal pathogens. Several studies support the use of tetracyclines in the treatment of localized juvenile periodontitis. Penicillins such as amoxicillin are effective in vitro against most periodontal pathogens but have limited efficacy due to the presence of beta-lactamases in gingival fluid. Amoxicillin/clavulanate potassium (Au) has proven effective in treating adult refractory periodontitis characterized by a Gram-positive flora. Metronidazole is an effective adjunct in adult periodontitis associated with high numbers of "black-pigmented Bacteroides" ad spirochetes. A combination of metronidazole and amoxicillin produces a synergistic effect against A. actinomycetemcomitans and has been shown to be effective at eliminating this organism. Clindamycin is an effective adjunct in the treatment of adult refractory periodontitis associated with a predominantly Gram-negative flora. The use of macrolides, quinolones, and combinations of antibiotics is discussed. Clinical studies do not support the use of systemically-administered antibiotics in routine adult periodontitis. Clinical studies do, however, support the use of antibiotics in the treatment of specific periodontal diseases. PMID- 8410617 TI - A role for antibiotics in the treatment of refractory periodontitis. AB - Refractory periodontitis is considered by many investigators to be a separate disease entity that is descriptive of a particular patient who has multiple sites, rather than a few individual sites, that do not respond to conventional periodontal treatment modalities. Such patients continue to demonstrate loss of attachment and alveolar bone despite frequent periodontal treatment which includes surgical intervention, scaling and root planing, and often systemically administered tetracycline. Controlled clinical studies have demonstrated that both clindamycin-hydrochloride and amoxicillin/clavulanate potassium (Au) are beneficial when used in conjunction with periodontal scaling. Gordon et al. found improvements in attachment levels, inflammation, suppuration, and a decrease in pocket depths for up to 2 years following a 7-day course of clindamycin given in conjunction with a full-mouth scaling. The incidence of disease activity decreased from an annual rate of 8% of all sites prior to antibiotic treatment to 0.5% after treatment. Magnusson, reporting on a similar group treated with a 14 day course of Au, found an average loss of attachment of 2.2 mm and an increase in pocket depth of 1.5 mm in sites demonstrating disease progression prior to antibiotic treatment. At 3 months post-antibiotic therapy, these sites had regained an average of 2 mm of attachment and pocket depths had decreased an equivalent amount. Both attachment levels and pocket depths remained relatively stable for up to 12 months post-therapy. In an ongoing study, 30 subjects with refractory periodontitis were treated with either clindamycin or Au in conjunction with scaling or scaling plus a placebo. Prior to antibiotic treatment, but while being scaled at 3-month intervals, sites with disease activity lost an average 2.4 mm of attachment. At 3 months post-treatment, the clindamycin-treated group showed an average gain of 2.1 mm, the Au-treated group gained 1.9 mm, and the scaling group gained 1.4 mm in attachment. The clindamycin group remained relatively stable for up to 21 months and the Au group remained stable for about 15 months without additional treatment. Five of the 6 subjects treated with scaling alone required additional treatment within 9 months. Preliminary analyses have indicated that at least two patterns or rates of attachment loss may be associated with refractory periodontitis and that each pattern may be indicative of a different microflora. The pattern associated with a relatively rapid loss of attachment was characterized by a Gram-negative flora which contained spirochetes, P. intermedia, and Fusobacterium species.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410618 TI - Controlled-release local delivery antimicrobials in periodontics: prospects for the future. AB - New knowledge about the microbial etiology of periodontal diseases emerged in the 1970s and 1980s and led to widespread interest in the use of antimicrobial agents to treat periodontitis. The controlled-release delivery of antimicrobials directly into the periodontal pocket has received great interest and appears to hold some promise in periodontal therapy. Some techniques for applying antimicrobials subgingivally, such as subgingival irrigation, involve local delivery but not controlled-release. Controlled-release local delivery systems, in which the antimicrobial is available at therapeutic levels for several days, have been evaluated in several forms and using different antimicrobials. Although most studies with such systems have focused on drug delivery kinetics and "proof of principle" evaluations, some controlled clinical trials have recently been reported. The most widely tested system, monolithic tetracycline-containing fibers, has shown significant clinical benefit when used alone as compared to no subgingival therapy. Similarly, controlled trials involving chlorhexidine strips used subgingivally every 3 months in place of routine supportive periodontal therapy have shown significant clinical benefit for up to 2 years. Although these data are now emerging, many questions concerning the optimal use and role of this therapy in clinical practice remain. This review attempts to summarize and interpret current data and to outline key remaining questions that must be addressed as this technology is transferred into clinical practice. PMID- 8410619 TI - The role of the host response in periodontal disease progression: implications for future treatment strategies. AB - In this paper, we review the relevant aspects of host responses in periodontal diseases as we understand them today. Discussion will focus on neutrophil function, lymphocytes and the immune response, macrophage function, cytokines and complement, fibroblasts and growth factors, and regeneration. Recent literature and concepts will be presented with an emphasis on future directions and application to treatment regimens. PMID- 8410620 TI - Specific antibodies and their potential role in periodontal diseases. AB - Periodontal diseases are thought to result from inflammatory responses to bacterial challenges in the gingival crevicular area. Antibodies are a major host protective mechanism in many bacterial infections. Consequently, the antibody responses to suspected periodontal pathogenic bacteria have been extensively measured as to their relationship to diseases and specificity for suspected pathogens associated with progressing disease sites. Recently, studies on the bacterial immunogen characterization, antibody-subclass identification, and antibody biological capabilities have been reported. Although increased antibody levels to certain suspected periodontal pathogens were associated with periodontal diseases in humans, little evidence exists as to the role of these antibodies in the infectious process. In vivo experiments in animals indicated that specific antibodies against certain suspected periodontal pathogens were associated with suppression of bacterial colonization, limiting the spread of infection, and a decrease in alveolar bone loss. However, in vitro as well as in vivo experiments suggested that phagocytic cells are required for efficient bactericidal activity of antibodies and that the presence of other sensitized immune cells may either have inhibited or enhanced the infectivity of certain periodontal pathogens. Possible explanations for the observed inconsistencies are presented and the potential for utilization of specific anti-periodontal pathogen responses in the understanding and prevention of diseases is discussed. PMID- 8410621 TI - Blocking periodontal disease progression by inhibiting tissue-destructive enzymes: a potential therapeutic role for tetracyclines and their chemically modified analogs. AB - Tetracyclines (TCs) have wide therapeutic usage as antimicrobial agents; these drugs (e.g., minocycline, doxycycline) remain useful as adjuncts in periodontal therapy. However, TCs also have non-antimicrobial properties which appear to modulate host response. In that regard, TCs and their chemically-modified analogs (CMTs) have been shown to inhibit the activity of the matrix metalloproteinase (MMP), collagenase. The activity of this enzyme appears crucial in the destruction of the major structural protein of connective tissues, collagen. Such pathologic collagenolysis may be a common denominator in tissue destructive diseases such as rheumatoid and osteoarthritis, diabetes mellitus, bullous dermatologic diseases, corneal ulcers, and periodontitis. The mechanisms by which TCs affect and, possibly, diminish bone resorption (a key event in the pathogenesis of periodontal and other diseases) are not yet understood. However, a number of possibilities remain open for investigation including the following: TCs may 1) directly inhibit the activity of extracellular collagenase and other MMPs such as gelatinase; 2) prevent the activation of its proenzyme by scavenging reactive oxygen species generated by other cell types (e.g. PMNs, osteoclasts); 3) inhibit the secretion of other collagenolytic enzymes (i.e. lysosomal cathepsins); and 4) directly affect other aspects of osteoclast structure and function. Several recent studies have also addressed the therapeutic potential of TCs and CMTs in periodontal disease. These drugs reduced excessive gingival collagenase activity and severity of periodontal breakdown in rats infected with Porphyromonas gingivalis and in diabetic rats. Furthermore, the latter drug (CMT) was not associated with the emergence of TC-resistant microorganisms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410622 TI - Blocking periodontal disease progression with anti-inflammatory agents. AB - Recent advances in the identification of specific immuno-inflammatory pathways of periodontal disease have encouraged investigators to attempt to modulate some of these host responses in an attempt to slow the periodontal disease process. Some of the best known mediators of these immuno-inflammatory pathways are prostaglandins. The action of prostaglandins can be inhibited through the use of non-steroidal anti-inflammatory drugs (NSAIDs). This review examines research over the last two decades during which the effect of several NSAIDs on the progression of gingival inflammation and alveolar bone loss was explored. PMID- 8410623 TI - [Propolis extract. I. Acute toxicity and determination of acute primary cutaneous irritation index]. AB - In first part, the aim of this work is to study the orally subacute toxicity of a propolis extract on conscious mice. LD50 is more than 7.34 g/kg propolis extract: this product is not toxic. In second part, cutaneous primary irritation of several products: excipients and propolis extract alone or in ointments, is evaluated in the rabbit. The cutaneous reactions after reiterated applications for 14 days are observed. The index of cutaneous primary irritation is calculated by evaluation of the erythematous and oedematous lesions. The propolis extract is non irritant. PMID- 8410624 TI - [Propolis extract. II. Wound healing the the rat and rabbit]. AB - This work is related to wounds healing properties of a propolis extract. In first study on the Albinos Rabbit, the activity of a propolis extract is compared with these of a Peru balsam. Optimal concentrations of them in ointments are evaluated by applications on deep cutaneous scarifications. In order to go further into details, we have chosen in second part, another assay on the Rat, allowing the obtention of deeper wounds; By this way, more complete quantification of retained parameters and a better appraising of the wounds healing process evolution are possible. PMID- 8410625 TI - X-ray diffraction study of N-cyclopropylcarbonyl-2-(7-methoxy-1-naphthyl) ethylamine, a highly potent and selective melatoninergic agonist. PMID- 8410626 TI - [Oral emulsions: preparation and bioavailability]. AB - Although oral L/H emulsions are rare, they constitute interesting pharmaceutical forms. Such systems are easy to administer, are good to protect a fragile lipophilic active substance and are able to modify drug release. After a recall on the principal components frequently used in these forms, such as oils surface active agents and active drugs, the mechanism of bioavailability and influencing factors are detailed. Finally, evaluation methods in order to analyse in vitro release are discussed. PMID- 8410627 TI - [Drug-induced syndrome of inappropriate secretion of antidiuretic hormone (SIADH). Review of literature]. AB - The syndrome of inappropriate antidiuretic hormone secretion (SIADH) is caused by an incapacity of plasmatic hypotonia to inhibit antidiuretic hormone (ADH) secretion. This phenomenon leads to a water retention and an expanded extracellular fluid volume, and secondary to a dilution of plasmatic sodium and also to a renal loss of sodium. The SIADH is a relatively rare syndrome, which may occur in various circumstances: central nervous system diseases, cancers, infections... Sometimes, it may be observed as a drug-related side effect: carbamazepine, chlorpropamide, antidepressors, anticancer drugs are particularly involved. PMID- 8410628 TI - [Reaction and interactions of drugs]. PMID- 8410629 TI - [The histology of the tegumentary system]. PMID- 8410630 TI - [Percutaneous penetration of drugs]. PMID- 8410631 TI - [Excipients and new topical forms in dermatology]. PMID- 8410632 TI - [From herpes to zona]. PMID- 8410633 TI - [Scalp disorders]. PMID- 8410634 TI - [Impact of the environment on the skin]. PMID- 8410635 TI - [Brown spots]. PMID- 8410636 TI - [Common superficial cutaneous mycoses]. PMID- 8410637 TI - [Psoriasis]. PMID- 8410638 TI - [Dry skin in the child]. PMID- 8410639 TI - [Intolerance reactions to cosmetics]. PMID- 8410640 TI - An uncertain future. PMID- 8410641 TI - Nursing care of a child with meningococcemia. AB - This case presentation will discuss the pathophysiology of a child in septic shock due to Neisseria meningitidis. The most prevalent nursing care concerns of this case encountered during the pediatric intensive care unit (PICU) and during the general floor stay will be addressed. The nursing skill required for identifying problems and planning care that clearly fall under the nursing domain also will be covered. In addition, the complexities of this case demonstrate that collaboration between the PICU nurse and the general pediatric nurse is imperative for successful patient outcome. A.W. was a 5 1/2-month-old infant transported to our PICU from a referral hospital in the state. Diagnosis at time of admission was meningococcemia, disseminated intravascular coagulopathy, septic shock, respiratory failure, and purpura fulminans. There was a 2- to 3-day history of a runny nose, cough, and vomiting. On the day of admission, A.W. had three seizures and developed a fever and a purpuric rash. PMID- 8410642 TI - Caring for chronically ill children at home: factors that influence parents' coping. AB - This study examined 29 parents' perceptions of the stressfulness of copying with their chronically ill child's home care. The parents were primarily responsible for care regimes that represented a continuous range of caregiving burden. Data were obtained using the Clinician's Overall Burden Index (COBI) (Stein & Jessop, 1982), the Coping Health Inventory for Parents (CHIP) (McCubbin & Patterson, 1981), visual analogue scales, and home interviews with parents. Parents described many dimensions of caregiving burden. Increased caregiving burden was associated with greater stressfulness and the use of fewer helpful coping strategies. Three clusters of coping strategies were most helpful: (a) using family support, (b) maintaining a positive outlook, and (c) ensuring that care was performed. The need for a reconceptualization of caregiving burden is discussed. PMID- 8410643 TI - "High-tech" home care for children with chronic health conditions: a pilot study. AB - The purposes of this study were to pilot test the instrument and methods for a major study of caregivers of children who are technology dependent and to test constructs proposed for the major study in a discrete sample. The major study will be implemented in 13 cities in the United States. The pilot was conducted in three small- to moderate-size metropolitan areas in the Southeast and Midwest. Seventy-three primary caregivers provided data during a structured telephone interviews. The children represented four groups of children dependent on technology as defined by the Office of Technology Assessment (OTA) (1987). The findings supported the methodology and the viability of the instrument. Analysis suggests that caring for a child who is technology dependent affects family functioning and increases family stress levels in some families. The financial burden is also significant. It is suggested that nurses can be a critical source of support and information to these families. PMID- 8410644 TI - The information and support needs of mothers of premature infants. AB - This research was a descriptive study that surveyed the support needs of a nonrandom convenience sample of 56 mothers of high-risk premature infants. The results indicated that these mothers' received information similar to that given to mothers of healthy full-term newborns and information that they wanted on the "colicky" or crying infant, noisy breathing, "spitting up," infant behavior, infant illness, and prematurity was not given to them. Mothers who reported that they needed more information and did not receive it were more anxious and less confident in caring for their infant. The majority of the mothers felt well supported, although many of the mothers found the infant's first postdischarge week difficult. PMID- 8410645 TI - Preterm infants' responses to taste/smell and tactile stimulation during an apneic episode. AB - A nonprobability sample of 14 nonventilated preterm infants, with a mean postconceptional age of 33.9 weeks, was examined to determine their responses to two interventions during apnea. The interventions included a traditional tactile stimulation of moderate shaking applied to the infant's leg and an experimental oral intervention consisting of taste, smell, and oral tactile stimulation. Infants served as their own control. Each infant received four randomly assigned trials (two of each intervention) when they experienced an apneic episode. The time interval for reinitiation of respiratory effort was significantly shorter after infants received the experimental stimulation (p = 0.0101). Behavioral state changed to alertness when the infants received the traditional tactile intervention yet remained unchanged when the experimental stimulation was administered during apnea (p = 0.0202). PMID- 8410646 TI - Sampling issues in parent-child nursing research: implications for nursing practice. PMID- 8410647 TI - Advanced practice in pediatric nursing. PMID- 8410648 TI - Airway support management: a teaching module. PMID- 8410649 TI - Preparing for cultural diversity through education. PMID- 8410650 TI - Effect of temporal focus on the recall of expectancy-consistent and expectancy inconsistent information. AB - This research examined the impact of temporal focus on the recall of information that is consistent or inconsistent with an expectation. A consistent pattern of results across 4 experiments indicates that when Ss' expectations are temporally unfocused, better memory for consistent information is observed. In contrast, when expectations are focused in time--that is, Ss know when the relevant events are likely to occur--recall for consistent and inconsistent information is more balanced. Experiment 4 tied these recall findings to the amount of processing devoted to consistent and inconsistent events. When expectations were temporally unfocused, processing time and recall was greater for the confirmatory information. When expectations were temporally focused, more equivalent processing time and recall of consistent and inconsistent information was observed. Discussion centers on the role of temporal focus as a determinant of whether an event is one-sided or two-sided. PMID- 8410651 TI - Beyond the actor's traits: forming impressions of actors, targets, and relationships from social behaviors. AB - Perceivers who observe social behaviors may form impressions not only of actors' traits but also of people as targets and of interpersonal relationships. In Study 1, Ss read about 4 individuals' behaviors under instructions to form actor-, target-, and relationship-based impressions. Ss then read additional behavioral information that they later tried to recall. Ss accurately perceived actor, target, and relationship effects in the presented information, and they better recalled subsequent behaviors that were consistent with all 3 types of impressions. In Study 2, Ss thought of 4 people they knew and judged how much each liked the other 3. These ratings revealed actor, target, and relationship effects as well as individual and dyadic reciprocity. Perceivers can form relatively accurate impressions of people as actors and as targets and accurate impressions of relationships between people, and these impressions influence memory for further behaviors. PMID- 8410652 TI - Perceptions of marital interaction among black and white newlyweds. AB - Perceptions of marital interactions were gathered from a representative sample of urban newlywed couples (199 Black and 174 White). A factor analysis of the reports found 6 factors common to husbands and wives: Disclosing Communication, Affective Affirmation, Negative Sexual Interaction, Traditional Role Regulation, Destructive Conflict, and Constructive Conflict. Avoiding Conflict was specific to men and Positive Coorientation was specific to women. Wives reported fewer constructive and more destructive conflict behaviors. Compared with Whites, Blacks reported more disclosure, more positive sexual interactions, and fewer topics of disagreement. They also more often reported leaving the scene of conflict and talking with others more easily than with the spouse. As hypothesized, perceptions that marital interactions affirm one's sense of identity strongly predicted marital well-being. Although regression analyses predicting marital happiness yielded few interactions with race or gender, those that are significant, coupled with race and gender differences in perceiving interaction, suggest taking a contextual orientation to the meaning of marital interaction. PMID- 8410653 TI - Two types of religious internalization and their relations to religious orientations and mental health. AB - Two types of religious internalization are conceptualized that are presumed to vary in their relative autonomy. Introjection represents a partial internalization of beliefs and is characterized by self- and other-approval-based pressures. Identification represents adoption of beliefs as personal values and is characterized by greater volition. These 2 types of internalization are compared conceptually and empirically with existing measures of religious orientation and are used to predict varied functional outcomes. Results in 4 independent Christian samples show systematic construct validities and relations with mental health and self-related outcomes. Also, evangelical teenagers are shown to be higher on both introjection and identification than controls. Results are discussed both in terms of prior approaches to the psychology of religion and the significance of internalization for personality functioning. PMID- 8410654 TI - Personality change in women and their partners. AB - Adjective Check List (ACL) data from 82 female participants in a longitudinal study and their male partners were used to investigate personality change between the early parental and postparental periods. In the early parental period, men were more competent, whereas women were more emotionally dependent and more facilitative in their interpersonal relations. In the postparental period, primarily because of the greater breadth and sharper gradient of ACL changes in women, these differences were attenuated or even reversed. The influence of the mother role, women's status level in work, whether partners were the same or different, and personality differences between the women's mothers and fathers were considered. PMID- 8410655 TI - Helping one kid at the time: serving in Somalia. Interview by Janice C Lemke.. PMID- 8410656 TI - Caring across cultures: short terms abroad. Learning to love in Zaire. PMID- 8410657 TI - Caring across cultures: short terms abroad. Bush nursing in Benin. PMID- 8410658 TI - Caring across cultures: short terms abroad. How six months in Papua New Guinea changed my life. PMID- 8410659 TI - Why do nurses volunteer? PMID- 8410660 TI - Teaching transculturally. PMID- 8410661 TI - Who can use this stuff? RACORSE of course! (Recycling, Allocation & Conservation of Operating Room Supplies & Equipment). PMID- 8410662 TI - Mission opportunities for nurses. PMID- 8410663 TI - Motorcycle missionary. PMID- 8410664 TI - Love the alien as yourself. PMID- 8410665 TI - Thinking Christianly about culture. PMID- 8410666 TI - Effect of dosing frequency on ZDV prophylaxis in macaques infected with simian immunodeficiency virus. AB - The effect of dosing frequency on zidovudine (ZDV) prophylaxis against simian immunodeficiency virus (SIV) infection was examined in long-tailed macaque monkeys (Macaca fascicularis). The results indicate that dosing frequency is extremely important for drug efficacy. The monkeys were divided into three groups based on dosing frequencies of 6-, 8-, or 12-h intervals. All were given a total daily dose of 100 mg/kg of ZDV. The drug was administered subcutaneously starting 24 h before SIV inoculation, and treatment continued for an additional 28 days. With the total daily dose held constant, ZDV was most therapeutic when administered at 12-h intervals, less effective at 8-h intervals, and least effective at 6-h intervals. These results indicate that early ZDV treatment based on infrequent but high dosages may increase the antiretroviral effect of the drug. These findings could serve as a model for ZDV chemoprophylaxis in humans. In cases involving accidental exposure to SIV or human immunodeficiency virus (HIV-1 or HIV-2), immediate, high-dosage therapies may be most therapeutic. PMID- 8410667 TI - Autoantibodies against beta 2-microglobulin-free HLA antigens in AIDS patients. AB - Serum samples from 88 human immunodeficiency virus (HIV)-positive drug addicts have been investigated for the presence of antibodies to both beta 2 microglobulin (beta 2m)-free and beta 2m-associated HLA class I molecules. Using HIV-negative drug addicts as background control, we found that none of the Centers for Disease Control (CDC) stage II, 9.1% of CDC III, 36.4% of CDC IV A, and 45.5% of CDC IV C1 patients had significant levels of autoantibodies competing with the binding of the monoclonal antibody specific for beta 2m-free HLA I (L31 mAb). Using the mAb 01.65, recognizing the beta 2m-associated form of HLA class I molecules, a similar percentage of positive samples was found in the CDC II, CDC III, and CDC IV A patient groups; conversely, the percentage of positive serum samples was lower in the CDC IV C1 group. A lower number of systemic lupus erythematosus serum samples and none of the specimens from healthy adult subjects or patients suffering from recurrent Epstein-Barr virus infections were positive in both assays. Our data demonstrate the existence of an ongoing HLA class I-specific autoimmune response during AIDS disease development, which probably reflects a molecular mimicry between autologous histocompatibility antigens and HIV components. The relationship between the prevalence of autoantibodies against beta 2m-free HLA class I and disease progression suggests a possible pathogenetic role of these antibodies in the induction of the HIV associated immune deficiency. PMID- 8410668 TI - High prevalence of anti-cardiolipin antibody, C1q-, C3d-, and mRF-IgG immune complexes, and anti-nuclear antibody in hemophiliacs irrespective of infection with human immunodeficiency virus type 1. AB - We investigated the prevalence of various autoantibodies [anti-cardiolipin antibody (aCL), lupus anticoagulant (LA), immune complexes (ICs), anti-nuclear antibody (ANA), and anti-deoxyribonucleic acid antibody (aDNA)] in hemophiliac individuals with (n = 50) and without (n = 42) infection by human immunodeficiency virus type 1 (HIV-1). The positivity rate for ANA was similar in both groups, and none of the patients was positive for LA and aDNA. aCL was positive in 35 of 50 (70%) HIV-1-positive hemophiliac individuals and 33 of 42 (79%) HIV-1-negative hemophiliac individuals. However, the majority of the aCL was revealed to be beta 2-glycoprotein I independent, thus corresponding to a syphilis type aCL that does not cause the so-called antiphospholipid syndrome. A total of 39 of the 45 HIV-1 positive hemophiliac individuals (87%) and 34 of 41 HIV-1-negative hemophiliac individuals (83%) had at least one type of IC [C1q-, C3d-, and/or murine monoclonal rheumatoid factor (mRF)- IgG]. The mechanism producing various autoantibodies in hemophiliac persons irrespective of their HIV 1 status is still unclear, but pathogens (e.g., HIV-1, hepatitis B, and hepatitis C) and alloantigens in the blood products that these patients require may be possible candidates. The clinical significance of the presence of these autoantibodies and the underlying mechanisms involved both need to be clarified further. PMID- 8410669 TI - Antibody response to measles and rubella vaccine by children with HIV infection. AB - To determine the immunogenicity of the measles and rubella components of the measles, mumps, and rubella virus (MMR) vaccine in human immunodeficiency virus (HIV)-infected children, we compared their response to that of uninfected controls. Sera were collected from HIV-infected patients and HIV seroreverters followed in our clinic and tested as close to 2 months post-MMR vaccination as possible. Specific IgG to both rubella and measles were measured by enzyme-linked immunosorbent assay. Of 20 children with HIV, 11 responded with adequate levels of antibody to measles. In the seroreverters, 12 of 13 responded. Of the measles responders, the median antibody level was significantly lower in the HIV-infected group than in the seroreverter group. In addition, HIV-infected responders tested at 9-15 months after vaccination demonstrated a significant decline in measles antibody levels. Although there was not a difference between the two cohorts in the proportion of patients who responded to the rubella component of the vaccine, there was a significant difference in the median antibody level of the responders of the two groups. We did not find a statistical difference in CD4 counts between responders and nonresponders. Alternate strategies will need to be established to prevent measles in HIV-infected children. PMID- 8410670 TI - Age at AIDS diagnosis for children with perinatally acquired HIV. AB - The human immunodeficiency virus (HIV) can be transmitted from mother to child in utero, during birth, or after birth through breast milk. While the majority of children born to HIV-positive mothers are not infected, almost all carry maternal antibodies. The number of maternal or perinatal exposures can be determined by screening all newborns for these antibodies, while maintaining the anonymity of mother and child. Combining newborn screening results with traditional surveillance data from New York City, we estimate that among maternally infected children, almost 14% will be diagnosed in the 1st year of life and, approximately 11-12% each year after, through age 7. This implies a median diagnosis age of 4.1 years, and suggests that in the majority of infected children, AIDS will not develop until after the 1st few years of life. We also examine the diagnosis age distribution for all children (infected and not) with a positive screen. We conclude that by approximately 7 years of age, 17% of children who test positive will be diagnosed with clinical AIDS, according to a yearly rate of > 2%. Based on these results, the perinatally acquired pediatric epidemic can be projected from the newborn screening data. PMID- 8410671 TI - Survival differences and trends in patients with AIDS in the United States. AB - The AIDS surveillance system maintained by the Centers for Disease Control and Prevention (CDC) provides a unique data base for estimating survival after a diagnosis of AIDS for the general AIDS population in the United States. Because patients enrolled in most AIDS clinical trial studies receive unusual medical care that may not be available to the general public and typically have relatively longer survival time, estimates obtained from these studies may not be of direct use in assessing the national health-care needs. Furthermore, such studies are usually of short duration and may not be very informative for long term health-policy planning. We present survival estimates obtained from the CDC surveillance data for the adult/adolescent AIDS population in the United States and compare their survival and trend in survival on gender, sexual behavior, and injection-drug use status. These estimates provide information for mortality risk after an AIDS diagnosis over a period of 8 years and for trend of survival during the period between 1983 and 1991. PMID- 8410672 TI - Survival after AIDS diagnosis in Washington State: trends through 1989 and effect of the case definition change of 1987. AB - Survival analysis was performed for AIDS cases diagnosed in Washington state from 1982 through 1989 and reported through October 31, 1991. No difference in survival time among diagnosis years 1987, 1988, and 1989 (p = 0.29) was found. Since September 1987, survival time was longest for cases with human immunodeficiency virus (HIV) wasting syndrome and HIV encephalopathy. Adjusted risk for death was significantly lower for these cases relative to all other cases (relative risk, 0.5; 95% confidence interval, 0.4-0.6). Explanations for the absence of continuing increase in survival time between 1987 and 1989 include changes in the frequency and timing of anti-HIV therapy. Longer survival time among cases diagnosed with HIV wasting or HIV encephalopathy is likely due to diagnosis earlier in the course of HIV disease. These results emphasize how changes over time in the definition of AIDS and evolving therapeutic standards may affect assessment of survival time when using surveillance data. PMID- 8410673 TI - No evidence of perinatal transmission of HTLV-II. AB - Two hundred twenty-eight plasma specimens collected over a 4-year period from 185 children aged 4 days to 10 years at risk for human immunodeficiency virus (HIV) infection were tested for antibody to human T-cell lymphotropic virus type II (HTLV-II) to determine perinatal transmission of this agent. One hundred three of the specimens were from 68 children whose mothers were intravenous drug users (IVDU). None of the children in the study were breast fed. No HTLV antibody was detected in any of the samples from children of non-IVDU mothers. However, HTLV II antibody was found in samples from 15 of 38 infants < 6 months old born to IVDU mothers. Later specimens taken at 8-16 months from six of these children were negative, suggesting passive maternal antibody, not infant infection. The 62 samples from 30 children 6 months to 7 years old from IVDU mothers were negative for HTLV antibody. The lack of HTLV antibody in the older children indicates that no vertical transmission had occurred. Based on the prevalence rate of maternal HTLV-II antibody found in the children < 6 months old, an HTLV-II infection rate of approximately 36% was projected for the IVDU mothers in the study. PMID- 8410674 TI - Evaluation of six rapid test kits for HIV-1 antibody detection. PMID- 8410675 TI - Risk assessment of HIV transmission among lesbians. PMID- 8410676 TI - Cigarette smoking, leukocyte profiles, and HIV-1 progression. PMID- 8410677 TI - CD8+ T cell counts up to 10 years from seroconversion. PMID- 8410678 TI - HIV-2 infection in Romania. PMID- 8410679 TI - Pharmacokinetics and pharmacodynamics of furosemide in protein-calorie malnutrition. AB - The influence of dietary protein deficiency on pharmacokinetics and pharmacodynamics of furosemide was investigated after i.v. bolus (1 mg/100 g) and oral (2 mg/100 g) administration of furosemide to male Sprague-Dawley rats fed on a 23% (control) or a 5% (protein-calorie malnutrition: PCM) protein diet ad lib. for 4 weeks. After i.v. administration, the mean values of CLR, Vss, and the percentages of dose excreted in 8-hr urine as furosemide were increased 81, 31, and 61%, respectively, in PCM rats when compared with those in control rats, however, CLNR was 54% decreased in PCM rats. The decreased CLNR in PCM rats suggested the significantly decreased nonrenal metabolism of furosemide. The urine volume per g kidney after i.v. administration was not significantly different between the two groups of rats although the amount of furosemide excreted in 8-hr urine per g kidney increased significantly in PCM rats. The diuretic, natriuretic, kaliuretic, and chloruretic efficiencies reduced significantly in PCM rats after i.v. administration. After oral administration, the extent of bioavailability increased considerably from 27.6% in control rats to 47.0% in PCM rats, probably as a result of decreased gastrointestinal and hepatic first-pass metabolism. This was supported by a tissue homogenate study; the amount of furosemide remaining per g tissue after 30-min incubation of 50 micrograms of furosemide with the 9000 x g supernatant fraction of stomach (42.4 vs. 47.9 micrograms) and liver (41.4 vs. 45.9 micrograms) homogenates increased significantly in PCM rats. No significant differences in CLR and t1/2 were found between the control and the PCM rats after oral administration. The 24-hr urine volume and the amount of sodium excreted in 24-hr urine per g kidney increased significantly in PCM rats, and this might be due to a significantly increased amount of furosemide reaching the kidney excreted in urine per g kidney. PMID- 8410680 TI - Terminal half-life. PMID- 8410681 TI - Variability in the renal clearance of cephalexin in experimental renal failure. AB - This study forms a part of an investigation into the extent to which the type of renal damage influences the renal clearance of drugs. We have already demonstrated an effect of different types of experimental renal failure (ERF) on the renal clearance of two cations: cimetidine, a drug that is filtered and secreted by the nephron, and lithium, which is filtered and reabsorbed by more than one segment of the nephron. In this report the renal clearance of cephalexin (CLCEX) is investigated, a drug that has a different mode of renal elimination, since it is filtered, secreted, and reabsorbed by the proximal tubules. The aim was to extend our earlier studies to an organic anion, and to provide an opportunity to evaluate the feasibility of using the renal clearance of N-1 methylnicotinamide (NMN) to predict the renal clearance of anionic drugs in renal failure. Different models of site-specific ERF have been developed in the rat; proximal tubular necrosis (induced by cisplatin), papillary necrosis (induced by 2-bromoethylamine), and glomerulonephritis (induced by sodium aurothiomalate or by antiglomerular basement membrane antibodies). Glomerular function (GFR) was assessed by the clearance of inulin (CLNULIN), and tubular function was assessed by the clearance of endogenous NMN (CLNMN). OUr results show that even if the models of ERF used were not absolutely site-specific, glomerular function and tubular function did not decrease to the same extent in the different ERF. Therefore, glomerulo-tubular imbalance existed, which is incompatible with the "intact nephron hypothesis," i.e., the site of the damage along the nephron and not only the degree of renal dysfunction, is a potential source of variability in the clearance of certain drugs. The renal clearance of cephalexin was estimated more accurately by CLNMN than GFR (r = 0.90). We conclude that the clearance of the endogenous cation NMN can be used to predict the renal clearance of drugs that are not only filtered by the glomeruli but also secreted and/or reabsorbed by the proximal tubules, and in the limited examples investigated appears to apply to both anionic and cationic compounds. In this respect the GFR alone was not an adequate parameter for the prediction of the renal clearance of such drugs. PMID- 8410682 TI - Classification of benzodiazepine hypnotics in humans based on receptor occupancy theory. AB - Benzodiazepine (BZP) hypnotics are now classified into four groups according to their plasma elimination rates: ultrashort-, short-, intermediate-, and long acting drugs. Since the specific binding affinities for the BZP receptor vary widely among the BZPs and their active metabolites, it may be more reasonable to correlate their pharmacological activities with the BZP receptor occupancy rather than with their plasma concentrations. The time courses of total plasma concentrations of BZPs and their active metabolites after a single oral administration were obtained from the literature, and their unbound concentrations (Cu) were calculated from the reported values of their plasma unbound fractions. The data of the receptor binding affinities of the BZPs, reported as dissociation constants (Kd) determined by in vitro binding experiments, were also obtained from the literature. Using these values, the time courses of receptor occupancies [Cu/(Kd + Cu) x 100%] were calculated for the various BZPs. A mutual competitive inhibition was considered in the case of the drugs that had active metabolites. Although plasma total and unbound concentration time profiles of the BZPs showed a wide variation, similar patterns were obtained for the time courses of the receptor occupancy among the BZPs in each group, indicating that the BZP hypnotics can be classified more conveniently based on receptor occupancy theory. PMID- 8410683 TI - Nonlinear protein binding and enzyme heterogeneity: effects on hepatic drug removal. AB - The kinetics of substrate removal by the liver and the resulting nonlinear changes in unbound fraction along the flow path at varying input drug concentrations were examined by a model simulation study. Specifically, we varied the binding association constant, KA, and the Michaelis-Menten constants (Km and Vmax) to examine the steady state drug removal (expressed as hepatic extraction ratio E) and changes in drug binding for (i) unienzyme systems and (ii) simple, parallel metabolic pathways; zonal metabolic heterogeneity was also added as a variable. At low KA, E declined with increasing input drug concentration, due primarily to saturation of enzymes; only small differences in binding were present across the liver. At high KA, a parabolic profile for E with concentration was observed; changes in unbound fraction between the inlet and the outlet of the liver followed in parallel fashion. Protein binding was the rate determining step at low input drug concentrations, whereas enzyme saturation was the rate-controlling factor at high input drug concentration. Heterogeneous enzymic distribution modulated changes in unbound fraction within the liver and at the outlet. Despite marked changes in unbound fraction occurring within the liver for different enzymic distributions, the overall transhepatic differences were relatively small. We then investigated the logarithmic average unbound concentration and the length averaged concentration as estimates of substrate concentration in liver in the presence of nonlinear drug binding. Fitting of simulated data, with and without assigned random error (10%), to the Michaelis Menten equation was performed; fitting was repeated for simulated data obtained with presence of a specific inhibitor of the high-affinity, anteriorly distributed pathway. Results were similar for both concentration terms: accurate estimates were obtained for anterior, high affinity pathways; an overestimation of parameters was observed for the lower affinity posteriorly distributed pathways. Improved estimations were found for posteriorly distributed pathways upon inhibition with specific inhibitors; with added random error, however, the improvement was much decreased. We applied the method for fitting of several sets of metabolic data obtained from rat liver perfusion studies performed with salicylamide (SAM) (i) without and (ii) with the presence of 2,6-dichloro-4 nitrophenol (DCNP), a SAM sulfation inhibitor. The fitted results showed that SAM sulfation was a high-affinity high-capacity pathway; SAM glucuronidation was of lower affinity but comparable capacity as the sulfation pathway, whereas SAM hydroxylation was of lower affinity and lower capacity. PMID- 8410684 TI - Influence of barrier-crossing limitations on the amount of macromolecular drug taken up by its target. AB - Macromolecules (substitutive enzymes, polymeric prodrugs, immunotoxins, radiolabeled antibodies, or peptide hormones) are of interest in the treatment of several diseases. To reach the tissues, these macromolecular drugs have to cross the capillary wall, which represents an important transfer limitation. While pharmacokinetics usually studies the changes in drug concentration in different body compartments, analyzing the amount of drug gaining access to its target may be more relevant for assessing the efficiency of macromolecules than for low molecular mass drugs. To determine the influence of different parameters on the fraction of the injected dose gaining access to the pharmacologic target, we constructed pharmacokinetic models where two uptakes, both linear or nonlinear, work either in the same compartment (no transport limitation), or in compartments separated by a transport barrier. Numerical applications were carried out with parameters obtained either experimentally or from the literature. We conclude that it is of little use to increase the affinity (K(uptake)) of a macromolecular drug for its target when a transport limitation and an undesired elimination from the plasma space are both present. Likewise, an increase of the uptake (rate of uptake or maximal velocity) by the target is not very productive because permeability of the capillary wall is the factor limiting access of macromolecules to tissues. Maximal efficiency of therapeutic macromolecules could be achieved by increasing, where feasible, the transport across the barrier between the plasma and the target, and by preventing the undesired eliminations as much as possible. PMID- 8410685 TI - Generalized pharmacokinetic modeling for drugs with nonlinear binding: I. Theoretical framework. AB - The following integrodifferential equation is proposed as the basis for a generalized treatment of pharmacokinetic systems in which nonlinear binding occurs phi'(cu)c'u = -q(cu)+g * cu+f where cu identical to unbound plasma drug concentration, f identical to drug input rate, ' indicates the derivative of a function, and * indicates the convolution operation: (g * cu) (t) = integral of t0 g(t-u)cu(u) du. Possible physical interpretations of the functions q, g and f are: q(cu) identical to rate at which drug leaves the sampling compartment, g * cu identical to rate at which drug returns to the sampling compartment from the peripheral system (tissues that are kinetically distinct from the sampling compartment), and phi(cu) identical to amount of drug in the sampling compartment. The approach assumes that drug binding is sufficiently rapid that it may be treated as an equilibrium process. It may be applied to systems in which nonlinear binding occurs within the sampling compartment, i.e., in the systemic circulation or in tissues to which drug is rapidly distributed. The proposed relationship is a generalization of most existing models for drugs with nonlinear binding. It can serve as a general theoretical framework for such models or as the basis for "model-independent" methods for analyzing the pharmacokinetics of drugs with nonlinear binding. Computer programs for the numerical solution of the integrodifferential equation are presented. Methods for pharmacokinetic system characterization, prediction and bioavailability are presented and demonstrated. PMID- 8410686 TI - Extracellular pH signals affect rat vascular tone by rapid transduction into intracellular pH changes. AB - 1. Changes in extracellular pH (pHo) are known to produce large effects on vascular tone, but the mechanisms involved are not understood. As changes in intracellular pH (pHi) can also affect vascular tone, we have investigated the effects of changing pHo upon both pHi and tone. 2. Strips of rat mesenteric resistance vessels were loaded with the pH-sensitive fluorophore SNARF 1; thus tension and pHi could be simultaneously measured as pHo was altered. 3. Whenever pHo was altered there was a corresponding alteration of pHi. Furthermore, when pHo was changed the pHi change was more rapid than that reported to occur in other cells. The time to half-peak intracellular response was 38 +/- 4 s (n = 11). The induced pHi change was also less attenuated than in many other cells studied. Thus a ratio of 0.73 was obtained for the change in pHi per unit pHo change (n = 7). 4. An increase in pHi produced by an increase in pHo was accompanied by an increase in tension in the vascular strips. In other experiments pHi was increased at constant pHo by the addition of the weak base trimethylamine (40 mM). This also elevated tension in the strips. Conversely when pHo was changed while pHi was held at resting values, no change in vascular tone occurred. 5. It is concluded that the effects of pHo on vascular tone are due to the induced change in intracellular pH, and that the vascular smooth muscle cell is functionally well adapted to respond to changes in tissue pH, thereby allowing blood flow to a tissue to be rapidly altered to meet changing needs. PMID- 8410687 TI - The effect of non-quantal acetylcholine release on quantal miniature currents at mouse diaphragm. AB - 1. The amplitude and exponential decay time constant of miniature endplate currents (MEPCs) were measured in mouse diaphragms treated with anti cholinesterase under conditions known to modulate non-quantal acetylcholine (ACh) release. 2. Anti-cholinesterase prolonged MEPC decay and the extent of this initial prolongation was not influenced by non-quantal release. When non-quantal release was present, the decays of MEPCs became increasingly faster over several hours. This increased decay did not occur in the absence of non-quantal release. 3. Potentiation of the non-quantal release by zero Mg2+ and 1 x 10(-5) M choline, on the other hand, led to acceleration of MEPC shortening. 4. Increase of temperature from 15 to 26 degrees C and the presence of the desensitization promoting drug proadifen (5 x 10(-6) M) accelerated the rate of MEPC shortening. 5. These observations are consistent with increased receptor desensitization due to non-quantal release. Repetitive binding of ACh to postsynaptic receptors which prolongs the time course of MEPC in anti-cholinesterase-treated endplates leads to progressive desensitization in the presence of non-quantal release and to the subsequent shortening of the quantal responses. PMID- 8410688 TI - Primary afferent depolarization of myelinated fibres in the joint and interosseous nerves of the cat. AB - 1. Changes in the excitability of the intraspinal terminals of fibres in the posterior knee joint and interosseous nerves were used as a measure of primary afferent depolarization (PAD) which is associated with presynaptic inhibition of transmission from afferent fibres. These were estimated from changes in the intensity of electrical stimuli required to activate the fibres in 50% of trials. In order to avoid the inclusion of group I muscle afferents which contaminate the joint and interosseal nerves, the analysis was restricted to fibres conducting at less than 75 m s-1 and/or displaying patterns of PAD which differed from those of group Ia and Ib muscle afferents in lower lumbar segments of anaesthetized cats. PAD was evoked by electrical stimulation of ipsilateral hindlimb nerves. 2. PAD of fibres in the posterior knee joint nerve was induced from group I (Ia and Ib) and group II muscle afferents and cutaneous afferents but not by stimulation of the joint or the interosseous nerves. The most effective stimuli were those applied to the superficial peroneal, sural, quadriceps and posterior biceps and semitendinosus nerves. 3. PAD of fibres in the interosseous nerve was also induced by stimulation of group I (Ia and Ib) and group II muscle afferents and cutaneous afferents and, in addition, by stimulation of joint and interosseous nerves. The most effective stimuli were those applied to the superficial peroneal, quadriceps, flexor digitorum longus and posterior biceps and semitendinosus nerves. 4. Individual fibres of the joint and the interosseous nerves were depolarized by only some of the conditioning stimuli. Even the most effective stimuli did not produce PAD in all of the fibres tested. Individual fibres of the joint and the interosseous nerves were depolarized by diverse combinations of afferents of different functional types and of different peripheral nerves. The differences in the sources of PAD were not associated with the conduction velocities and hence are unlikely to be related to differences in the receptor origin of the tested fibres. The diversity in the sources of PAD of individual fibres is interpreted as providing a high degree of differentiation in the control of transmission from receptors in joints and interosseal membranes. PMID- 8410689 TI - The effect of reversible cooling of cat's primary visual cortex on the responses of area 21a neurons. AB - 1. Responses of sixty-four neurons in cortical area 21a were studied with areas 17 and 18 reversibly deactivated by cooling. From anatomical studies, most of area 21a input in the cat originates from these primary areas with no input from the A laminae of the lateral geniculate nucleus. 2. Both responses and spontaneous activity of all sixty-four area 21a neurons were markedly reduced when primary areas were cooled. In sixteen cells the responses were totally blocked. Temperatures of primary cortex required to produce total blockade varied between 25 and 4.5 degrees C. 3. The effect of cooling the primary visual cortex on the shape of orientation tuning curves was analysed in thirteen neurons from area 17 and in fifty-eight neurons from area 21a. In both areas, the width of the curve, when measured at its half-height, was preserved even when spike activity was reduced to below 10% of the original level. All neurons also retained their original directional preferences during cooling of the primary visual cortex. 4. The responsiveness of forty-seven neurons from area 21a was tested after rewarming of primary cortex. All but one neuron recovered their initial responsiveness within half an hour of restoring physiological temperature of primary visual cortex. 5. The results of the present study give an indication of the extent to which area 21a is sequentially related to the primary visual cortex in the processing of information. PMID- 8410690 TI - M2 muscarinic receptor-mediated inhibition of the Ca2+ current in rat magnocellular cholinergic basal forebrain neurones. AB - 1. The actions of muscarinic agonists and antagonists upon the Ca2+ current (ICa) in acutely dissociated magnocellular cholinergic basal forebrain neurones from 11 to 14-day-old postnatal rats were studied using the whole-cell patch-clamp technique. 2. In all cells studied, muscarinic agonists inhibited a transient component of high-voltage-activated (HVA) current, but had no effect upon the low voltage-activated (LVA) current. The mean IC50 values for ACh and oxotremorine methiodide (oxo-M), obtained from non-cumulative dose-response curves, were 204 and 363 nM respectively. Superfusion with the K+ channel blocker, tetraethylammonium chloride (TEA; 30 mM) shifted the ACh dose-response curve to the right giving an IC50 value of 22:9 microM. 3. Pirenzepine (0.1-1 microM) and methoctramine (0.03-0.3 microM) produced parallel shifts to the right in the agonist dose-response curves. Schild analysis of the agonist dose ratios yielded pA2 (negative log of the apparent dissociation constant KB) values for pirenzepine and methoctramine of 6.8 and 8.2 respectively, indicating the involvement of an M2 receptor subtype. 4. In the presence of GTP-gamma-S (10-500 microM) in the patch pipette, the agonist-induced inhibition of ICa became irreversible. Dialysis with GDP-beta-S (1 mM) abolished all agonist-induced inhibition of the Ca2+ current. The agonist-induced inhibition of ICa was totally blocked by pretreatment with pertussis toxin (500 ng ml-1) but unaffected by preincubation with cholera toxin (500 ng ml-1). Superfusion with 8-bromo cAMP (0.5-1 mM) did not mimic or prevent the effect of agonist application. 5. Inhibition of the Ca2+ current by muscarinic agonists was only partially blocked by omega-conotoxin GVIA (omega-CgTX GVIA), with approximately 46% of the agonist sensitive current being resistant to omega-CgTX GVIA. Both the agonist- and omega CgTX GVIA-sensitive components of the current were abolished following maximal inhibition of ICa by GTP-gamma-S. 6. These results indicate that inhibition of the Ca2+ current by muscarinic agonists is mediated via an M2 muscarinic receptor coupled to a pertussis toxin-sensitive G-protein. The possible modulation of multiple HVA Ca2+ channels by muscarinic agonists and the role that these receptors may play in presynaptic modulation of neurotransmitter release are discussed. PMID- 8410691 TI - Convergence in segmental reflex pathways from nociceptive and non-nociceptive afferents to alpha-motoneurones in the cat. AB - 1. Reflex interaction between nociceptive and non-nociceptive segmental afferents was investigated by testing for spatial facilitation of postsynaptic potentials (PSPs) in alpha-motoneurones recorded in anaemically decapitated, high spinal cats. Nociceptive segmental afferents were activated by applying noxious radiant heat to the skin. Non-nociceptive skin mechanoreceptors were activated by puffs of air. Non-nociceptive skin, joint and group I-III muscle afferents were stimulated by electrical pulses delivered to various nerves. 2. Conditioning by stimulation of nociceptive afferents facilitated transmission in various ipsilateral segmental pathways. Such spatial facilitation occurred in both excitatory and inhibitory pathways. Pathways that were facilitated included those activated by low to medium threshold cutaneous afferents, joint afferents, and group Ib and II muscle afferents. 3. In contrast, monosynaptic EPSPs evoked by stimulating ipsilateral group Ia muscle afferents did not show spatial facilitation but rather inhibition during conditioning stimulation of nociceptive afferents. Spatial facilitation of reciprocal group Ia IPSPs was rare and small if it occurred. 4. Pathways activated by cutaneous and group II muscle afferents were depressed by contralateral stimulation of nociceptive afferents. 5. We conclude that spatial facilitation observed between nociceptive and non nociceptive afferents results from a convergence of inputs on common interneurones in the reflex pathways to alpha-motoneurones. Therefore nociceptive afferents have to be regarded as constituents of flexor reflex afferents (FRAs) and may add a specific nocifensive function to the FRA system. PMID- 8410692 TI - Myofilament sliding per ATP molecule in rabbit muscle fibres studied using laser flash photolysis of caged ATP. AB - 1. To estimate the distance of myofilament sliding per ATP molecule hydrolysed during the actin-myosin interaction in muscle, single glycerinated fibres prepared from rabbit psoas muscle were made to shorten under very small external loads (< or = 0.0005 maximum isometric force (Po), at 20-22 degrees C) by the laser flash photolysis of caged ATP (P3-1-(2-nitro) phenylethyladenosine 5' triphosphate), a biologically inert and photolabile precursor of ATP. The laser flash-induced fibre shortening was recorded with a high-speed video system at 200 frames s-1. 2. Following the photochemical release of 75-300 microM ATP, the fibres shortened uniformly along the fibre length not only at the level of fibre segments but also at the level of sarcomeres. The fibres did not shorten appreciably in response to 50 microM ATP. 3. The initial velocity of the laser flash-induced fibre shortening increased with increasing concentration of released ATP, being 0.05 +/- 0.01, 0.12 +/- 0.04, 0.23 +/- 0.04, 0.38 +/- 0.03 and 0.95 +/- 0.08 microns s-1 (half-sarcomere)-1 (means +/- S.E.M., n = 10) with 75, 100, 150, 200 and 300 microM ATP, respectively. 4. The distance of the laser flash-induced fibre shortening also increased with increasing concentration of released ATP, being 10 +/- 2, 25 +/- 5, 65 +/- 7, 100 +/- 10 and 180 +/- 20 nm (half-sarcomere)-1 (means +/- S.E.M., n = 10) with 75, 100, 150, 200 and 300 microM ATP, respectively. 5. Comparison of the initial shortening velocities of the laser flash-induced shortening with the force-velocity relation of maximally Ca(2+)-activated fibres indicated the presence of considerable internal resistance against myofilament sliding following release of ATP. The initial velocity of shortening following the release of 300, 150 and 75 microM ATP was equal to the shortening velocity of maximally Ca(2+)-activated fibres under an external load of 0.55, 0.93 and 0.98 Po respectively. 6. These results suggest that, under nearly isometric conditions, the distance of myofilament sliding per ATP molecule hydrolysed is about 10 nm in each half-sarcomere. PMID- 8410693 TI - Kinetics of ATP-induced Ca2+ transients in cultured pig aortic smooth muscle cells depend on ATP concentration and stored Ca2+. AB - 1. Single cultured pig aortic smooth muscle cells were studied using fura-2 and dual excitation wavelength microfluometry. 2. Extracellular ATP in micromolar concentrations induced a transient increase of [Ca2+]i due to Ca2+ release from internal stores. In the same concentration range application of ATP resulted in an increase of intracellular inositol phosphate level. 3. In a medium range of ATP concentrations (2-10 microM) the Ca2+ signal was oscillating, whereas at higher and lower concentrations only a Ca2+ transient with a single peak was elicited. 4. The rank order of potency for the tested purine and pyrimidine nucleotides was: UTP > ATP > ADP >> AMP = adenosine = alpha,beta-methylene ATP = 0. The response to the nucleotides could be abolished by the P2-purinoceptor antagonist suramin. 5. The latency between agonist application and onset of the Ca2+ transients as well as their amplitude and rate of rise are dependent on ATP concentration. 6. Removal of Ca2+ from the extracellular solution led to a progressive decrease of amplitude and prolonged latency of the Ca2+ transients. This shows that depletion of the Ca2+ stores affects kinetics of the ATP-induced Ca2+ release. 7. The inorganic Ca(2+)-influx blockers Ni2+ and Co2+ affected amplitude and latency in a manner similar to Ca2+ removal, while the Ca2+ antagonist nifedipine was ineffective up to a concentration of 10(-6) M. 8. These results reveal a dual dependency of the InsP3-induced Ca2+ release on agonist concentration and filling state of the Ca2+ stores, which supports the hypothesis of a feedback amplification between InsP3 and released Ca2+. PMID- 8410694 TI - Na(+)-Ca2+ exchange current in latent pacemaker cells isolated from cat right atrium. AB - 1. Single latent pacemaker cells were isolated from cat right atrium, and studied in a whole-cell configuration using a nystatin-perforated patch recording method. The nystatin method avoids alterations in intracellular Ca2+, cellular constituents and run-down of ionic currents. 2. Depolarizing voltage clamp pulses from -40 mV elicited L-type Ca2+ current (ICa) that exhibited an initial rapid phase of inactivation followed by a secondary slower inward current component that decayed over about 100 ms. The secondary inward component appeared as a slowly decaying inward tail current following short (10-40 ms) depolarizing clamp steps. 3. Slowly decaying inward currents were abolished by internally dialysing pacemaker cells with 2 mM EGTA using a ruptured patch recording method. Inward tail currents were also abolished by exposure to 1 microM ryanodine and significantly decreased by replacing 85% of external Na+ with lithium, without effect on peak ICa. These findings identify a Na(+)-Ca2+ exchange current (INa Ca) that is mediated by sarcoplasmic reticulum (SR) Ca2+ release. 4. Properties of INa-Ca and ICa differed significantly: (i) ICa exhibited a bell-shaped voltage dependence that peaked at 0 mV and decreased at more positive voltages. INa-Ca was maximal at -10 mV and remained relatively constant at more positive voltages; (ii) a paired pulse protocol showed that the time course of INa-Ca recovery (5 s) was significantly longer than that of ICa (2 s); (iii) cadmium (50 microM) induced an inhibition of ICa that did not correlate in time with changes in INa Ca. 5. The duration of depolarizing steps between 10 and 120 ms had no effect on the time course of INa-Ca tail currents. 6. Isoprenaline > or = 5 x 10(-8) M significantly increased peak ICa amplitude, peak INa-Ca amplitude, accelerated INa-Ca rate of decay and decreased the absolute time of INa-Ca decay. 7. Free running pacemaker action potentials were clamped during diastole at either -40 or -70 mV (maximum diastolic potential) for variable periods of time. At times between 0.2 and 1 s, INa-Ca exhibited a voltage-dependent increase in amplitude over time, i.e. INa-Ca recovered more rapidly from -70 mV than from -40 mV. At times > 2 s, INa-Ca exhibited a voltage-dependent decline in amplitude over time, i.e. from -40 mV INa-Ca decreased by 10% of maximum whereas from -70 mV INa-Ca decreased by 60% of maximum.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410695 TI - Gating properties of the cAMP-gated channel in toad olfactory receptor cells. AB - 1. Inside-out membrane patches were excised from different parts of isolated olfactory receptor cells of the toad, and the gating properties of cAMP-gated channels were investigated under low concentrations of divalent cations. 2. At +50 mV, an outward current was observed when 1 mM cAMP was applied to the cytoplasmic side of membrane patches excised from cilia. cGMP had a similar effect. The dose-response relation for the cAMP-induced current could be fitted with the Hill equation with a coefficient of 1.5 for both cAMP and cGMP, and half maximal concentration (K1/2) values of 19 microM (cAMP) and 16 microM (cGMP). 3. cAMP at low concentration (1 microM) induced step-like currents representing the opening of individual channels with a unitary conductance of about 30 pS. The I-V relation for the unitary events showed a weak outward rectification. 4. The relation between variance and mean current amplitude was well described with a parabola. Even at a saturating ligand concentration (1 mM cAMP) the current fluctuations did not disappear, indicating that fully liganded channels still switched between open and closed states. The maximum open probability was about 80% (+40 to +60 mV). 5. The current fluctuations at 1 mM cAMP were analysed with power spectral analysis. At +50 mV, frequency components lower than 10 Hz were well described with a Lorentzian function with a corner frequency of 1.7 +/- 0.2 Hz (+/- S.D.). In addition, higher flat frequency components were observed. At 50 mV the corner frequency became 0.6 +/- 0.1 Hz. 6. Membrane patches having a single cAMP-gated channel were obtained from the dendro-somatic membrane. These channels were very similar to the ciliary channels in electrophysiological characteristics. 7. The single cAMP-gated channel current showed flickering bursts that were interrupted with gaps. In saturating cAMP condition (1 mM), both burst- and gap-time histograms were fitted with single-exponential functions with time constants of 148 and 141 ms, respectively. 8. The channels were present at a high density of around 1750 microns-2 on the ciliary plasma membrane, as compared to 6 microns-2 on dendro-somatic membrane. PMID- 8410696 TI - Pathways through the intercellular clefts of frog mesenteric capillaries. AB - 1. The three-dimensional ultrastructure of endothelial intercellular clefts of frog mesenteric capillaries of known hydraulic permeability (Lp) has been investigated in the absence and presence of lanthanum ions as tracers of extracellular solute. 2. Experiments were carried out on the exposed mesenteries of pithed frogs and Lp of a chosen microvessel perfused with a Ringer solution containing serum albumin (10-40 mg ml-1) was determined. In some experiments the mesentery was fixed in situ with 2.5% glutaraldehyde immediately after Lp had been measured. In other experiments, measurement of Lp was followed by brief microperfusion (10-20 s) with a second Ringer solution containing 1% lanthanum nitrate as a tracer before in situ fixation of the tissue. The tissue was prepared for electron microscopy using standard techniques. The perfused capillary was identified in the block and serial transverse sections were cut along its length over regions where Lp had been measured. 3. In six capillaries where the tissues were fixed immediately after measurement of Lp, Lp had a mean value (+/- S.E.M.) of 4 (+/- 0.5) x 10(-7) cm s-1 (cmH2O)-1. Serial (30-40 nm) sections of these vessels revealed that a single short narrow region of the intercellular clefts ran almost continuously from section to section. Additional tight regions were regularly seen, but they usually extended for relatively few sections. In 13.36 microns of reconstructed cleft, there were three interruptions of the tight region of 0.14, 0.14 and 0.17 microns respectively. In the region of these discontinuities, the wide region was uninterrupted from luminal to abluminal surface. 4. Examination of the tight junction on a tilting stage revealed that the outer leaflets of the adjacent cells were not fused, but separated by a gap of mean width (+/- S.E.M.) 2.3 (+/- 0.1) nm. 5. In four capillaries perfused with lanthanum nitrate before fixation, mean Lp (+/- S.E.M.) was 6.5 (+/- 0.02) x 10(-7) cm s-1 (cmH2O)-1. Segments of intercellular clefts, totalling 23.56 microns in length, were reconstructed from serial sections and throughout these, electron-dense deposits of lanthanum were observed to fill the luminal parts of the intercellular clefts up to the tight region. Lanthanum deposits filled the entire cleft to the abluminal surface at eleven sites, which accounted for a length of 2.52 microns out of the 23.56 microns. Only five of these regions were delimited within a continuous series of sections and their mean length (+/- S.E.M.) was 0.16 (+/- 0.063) microns.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410697 TI - Ca(2+)-dependent block and potentiation of L-type calcium current in guinea-pig ventricular myocytes. AB - 1. The caged calcium compound nitr-5 has been used to investigate the response of the L-type calcium current (ICa) of guinea-pig ventricular cells to a rapid increase in the free intracellular calcium concentration ([Ca2+]i). 2. When 2 mM nitr-5 or 3 mM DM-nitrophen was loaded into cells via a patch pipette and photolysed during the decay phase of ICa, a partial block of the current developed within 75 ms. The block was reduced by increasing the pre-flash [Ca2+]i and enhanced by adding high concentrations of Ca2+ chelators to the pipette filling solution. 3. The photolysis-induced block was not suppressed in the presence of isoprenaline, suggesting a direct action of Ca2+ on the channels rather than a mechanism involving channel phosphorylation. 4. The most prominent effect of nitr-5 photolysis was a slow potentiation of ICa. When ICa was activated at frequencies between 0.05 and 0.7 Hz with various levels of pre-flash [Ca2+]i, peak ICa was approximately doubled in amplitude following photolysis. 5. At a stimulation frequency of 0.05 Hz, when nitr-5 was the only chelator present in the pipette, the time course of the potentiation was fitted by a single exponential with a time constant (tau P) of 2.7 min. When 1 mM CaCl2 was added to the pipette-filling solution, the time course of the potentiation was slowed (tau P = 6 min), although its amplitude was unchanged. With 12 mM BAPTA (a calcium chelator) added instead of CaCl2, the response was accelerated (tau P = 1.7 min). 6. Equimolar substitution of extracellular Ca2+ with Ba2+ significantly suppressed the flash-induced potentiation. The time course of the potentiation of the barium current, IBa (tau P = 1.9 min) was similar to that of ICa with BAPTA in the pipette. Potentiation of IBa was largely blocked in Ca(2+)-depleted cells when CaCl2 was omitted from the pipette. 7. When ICa was activated at frequencies of > or = 0.1 Hz, with 1 mM CaCl2 added to the nitr-5 (2 mM) in the pipette, the onset of the flash-induced potentiation was best fitted by two exponentials; one was similar to the single component seen at 0.05 Hz and the other was approximately one order of magnitude faster. The contribution of the faster component was positively correlated to the stimulation frequency. 8. The flash induced potentiation of ICa was suppressed in the presence of a supramaximal concentration of the beta-adrenergic agonist isoprenaline.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410698 TI - Quantal transmitter release at snake twitch and tonic muscle fibres during prolonged nerve terminal depolarization. AB - 1. Miniature endplate currents (MEPCs) were recorded in vitro from voltage clamped twitch and tonic muscle fibres in costocutaneous muscles of the garter snake, Thamnophis. Recordings were made from fibres in a control sodium containing solution and then during exposure to an isotonic potassium solution containing either 1.0 mM calcium and 4.2 mM magnesium or 3.6 mM calcium. The experiments were done at two levels of external calcium in order to demonstrate that the change in MEPC frequency was calcium dependent. During the initial exposure to the isotonic potassium solutions, the MEPC frequency was increased manyfold at both twitch and tonic fibres, but it declined progressively with continued exposure. MEPCs were recorded from both fibre types throughout a 20 h exposure to the isotonic potassium solution with 1 mM calcium, but no MEPCs were recorded at most twitch endplates after approximately 6 h in the isotonic potassium solution containing 3.6 mM calcium. In contrast, MEPCs were still present at tonic fibre endplates after 20 h in the isotonic potassium solution containing 3.6 mM calcium. 2. After 30 min in the isotonic potassium solution with 1 mM calcium, the MEPC amplitude recorded from both fibre types was approximately twice that in the control sodium-containing solution. At tonic endplates, the MEPC amplitude was also twofold greater in the isotonic potassium solution with 3.6 mM calcium than in sodium-containing solution. In contrast, after 30 min in the isotonic potassium solution containing 3.6 mM calcium, the MEPC amplitude at twitch endplates was similar to that in control solution. 3. In both fibre types, MEPC amplitude decreased progressively with continued exposure to the isotonic potassium solutions. The progressive decrease in MEPC amplitude was not due to a gradual decrease in postsynaptic sensitivity to acetylcholine. 4. The effects of high potassium were reversible as MEPCs were recorded at twitch fibre endplates in preparations which were returned to the control sodium containing solution after a 20 h exposure to the isotonic potassium solution containing 3.6 mM calcium. 5. Ultrastructural examination showed that after a 6 h exposure to the isotonic potassium solutions most nerve terminals innervating twitch fibre endplates were devoid of synaptic vesicles whereas at the same time many synaptic vesicles were present in nerve terminals innervating tonic fibre endplates. Surprisingly, numerous synaptic vesicles were present in nerve terminals innervating either fibre type in muscle preparations exposed to the isotonic potassium solutions for 20 h.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410699 TI - Sulphonylureas reduce the slowly inactivating D-type outward current in rat hippocampal neurons. AB - 1. Using intracellular recording in hippocampal slices, we have examined, in CA3 pyramidal neurons, the effects of sulphonylureas (blockers of ATP-sensitive K+ channels) on the slowly inactivating D-type K+ current (ID). 2. In the presence of TTX (1 microM) to block Na+ currents, ID had the following characteristics: activation by large depolarizing pulses from membrane potentials negative to -75 mV, slow inactivation kinetics, high sensitivity to 4-aminopyridine (4-AP, 3-40 microM), insensitivity to tetraethylammonium (TEA, 10 mM), Cs+ (3 mM) and carbachol (50 microM). 3. Applications of glibenclamide (10 microM) did not modify the input conductance of the cell, but reduced the amplitude of ID by 31.2 +/- 5.6% (n = 16), without altering its voltage dependence and inactivation kinetics. The effects were usually reversible. 4. Glibenclamide also reduced ID in the presence of TEA (10 mM), Cs+ (3 mM) and carbachol (50 microM), to block several K+ currents (IK, IC, IQ, IM), as well as kynurenate (1 mM) and bicuculline (10 microM) to block on-going synaptic currents mediated by activation of non-NMDA (N-methyl-D-aspartate) and GABA (gamma-aminobutyrate)-A receptors, respectively. 5. Comparable depressions of ID were produced by two other sulphonylureas: gliquidone (10 microM), 42.6 +/- 7.9% (n = 13) and tolbutamide (500 microM), 39.1 +/- 12.8 (n = 8). 6. It is concluded that, in the central nervous system, sulphonylureas can modulate K+ currents which are not generated by ATP-sensitive K+ channels. PMID- 8410700 TI - Effects of lanthanum at snake twitch and tonic muscle fibre endplates. AB - 1. The effects of 1 mM lanthanum on miniature endplate current (MEPC) frequency, amplitude, and decay time course were studied in voltage-clamped twitch and tonic muscle fibres in the garter snake, Thamnophis. 2. Lanthanum produced a marked increase in MEPC frequency in both fibre types. The maximum frequency in lanthanum was greater at twitch endplates than at tonic endplates although the increase in frequency relative to control levels was as great in tonic fibres as in twitch fibres. 3. In twitch fibres continually exposure to lanthanum, the frequency of MEPCs reached a peak value and then declined progressively until, after approximately 6 h, no MEPCs were recorded. In contrast, at tonic endplates exposed to 1 mM lanthanum, MEPC frequency remained elevated above control levels for periods greater than 20 h. 4. Lanthanum decreased the mean amplitude of MEPCs, skewed the amplitude distribution and increased MEPC duration at both twitch and tonic fibre endplates. 5. Ultrastructural analysis showed that after a 15 min exposure to 1 mM lanthanum, approximately half of the boutons innervating a twitch fibre contained fewer synaptic vesicles than boutons at control endplates, whereas nerve terminals innervating tonic fibre endplates were similar in appearance to those in control preparations. At endplates on both fibres, the postsynaptic membrane was more electron dense than that of control preparations. 6. Following a 6 h exposure to lanthanum, all nerve terminals innervating twitch endplates contained only a few synaptic vesicles and numerous intracellular deposits of electron dense material. The nerve terminals innervating tonic endplates still contained many synaptic vesicles, but the number appeared to be less than that of tonic terminals in untreated preparations. 7. The results demonstrate that lanthanum stimulates spontaneous quantal transmitter release from nerve terminals innervating either twitch or tonic fibres. However, the terminals innervating twitch fibres become depleted of synaptic vesicles, whereas this does not occur as readily in nerve terminals innervating tonic fibres. PMID- 8410701 TI - Co-existence and elimination of convergent motor nerve terminals in reinnervated and paralysed adult rat skeletal muscle. AB - 1. Experiments were carried out to determine whether neuromuscular synapse elimination can occur in skeletal muscle in the complete absence of conducted neural activity, using reinnervation of partially denervated adult muscle as a paradigm. Partially denervated rat lumbrical muscles were paralysed with a nerve conduction block applied to the sciatic nerve during regeneration of injured sural nerve motor axons. Both intact (lateral plantar nerve) and regenerating motor axons converging on the same muscle fibres were therefore inactive. 2. Paralysed muscles expressed prolonged twitch contractions, low tetanus-to-twitch ratios, prolonged synaptic potentials and marked post-tetanic potentiation of frequency of miniature endplate potentials compared with control muscles and neuromuscular junctions. 3. Isometric tension and intracellular recording data suggest that regenerating axons reinnervated more muscle fibres in paralysed muscles than in controls. A greater proportion of muscle fibres was polyneuronally innervated in the paralysed muscles, but significant numbers of muscle fibres acquired a mononeuronal innervation by regenerated, inactive motor nerve terminals. 4. The data suggest that muscle paralysis enhances the regeneration of motor axons when they grow into partially denervated muscles, but activity-independent competition may also be important in the mechanism of synapse elimination at neuromuscular junctions. The data further imply that when nerve endings expressing identical patterns of activity converge on a postsynaptic cell, Hebbian rules may not be sufficient to predict the outcome of the competition, contrary to specific postulates of the neurotrophic theory of development and maintenance of neural connections. PMID- 8410702 TI - The mechanism of ion transport by the Na(+)-Ca2+,K+ exchange in rods isolated from the salamander retina. AB - 1. Membrane currents caused by the operation of electrogenic Na(+)-Ca2+,K+ exchange were recorded from isolated rod outer segments under voltage-clamp using a whole-cell electrode. 2. Reversed mode exchange currents (Na+i-Ca2+o,K+o) were recorded with a high internal [Na+] and when both Ca2+ and K+ were present in the external solution. Omission of either Ca2+ or K+ completely suppressed both the reversed exchange current and the entry of Ca2+. 3. The charge transferred by the exchange per Ca2+ ion transported was identical in both forward and reversed modes. 4. The reversed exchange current declined as Ca2+ accumulated inside the outer segment, and the form of this decline was consistent with a first-order inhibition by internal Ca2+. 5. The reversed exchange current was increased e fold by a 230 mV depolarization over the range -51 to +29 mV. 6. The activation of reversed exchange by external Ca2+ was well described by first-order kinetics with a Michaelis constant, KappCao, of 34 microM in the presence of 20 mM external K+. KappCao was reduced by lowering external [K+], was increased by adding external Na+ and was unaffected by membrane potential. 7. External K+ also activated the exchange in a first-order manner with a Michaelis constant, KappKo, of 151 microM in the presence of 0.5 mM external Ca2+. KappKo was reduced by lowering external [Ca2+], increased by adding external Na+ and was unaffected by membrane potential. 8. When the level of internal Ca2+ was increased via reversed exchange, KappCao diminished in proportion to the reduction in the maximum current, but KappKo remained approximately constant. 9. These observations cannot be reconciled with simple models of the exchange in which ions bind simultaneously at opposite faces of the membrane before transport occurs. The results are broadly consistent with a consecutive model of the exchange in which unbinding of Na+ at either the external or the internal membrane surface is followed by binding of Ca2+ and then K+, and are fully reproduced by a model in which Ca2+ binds before all of the Na+ has dissociated from the exchange molecule. PMID- 8410703 TI - The mechanism by which cytoplasmic protons inhibit the sodium-calcium exchanger in guinea-pig heart cells. AB - 1. We recorded cardiac sodium-calcium exchange current (INa-Ca) in giant excised membrane patches obtained from cardiac myocytes of the adult guinea-pig. 2. Rapid changes in ion concentrations on the cytoplasmic side of the excised membrane patch were produced using a modified oil-gate bath. 3. Sodium-calcium exchange current was activated by step increases in sodium concentration on the cytoplasmic side of the membrane ([Na+]i), which led to an increase in outward INa-Ca to a new steady-state level. The [Na+]i required to half-maximally activate the sodium-calcium exchange current (K1/2) was 21 mM. 4. Step increases in cytoplasmic calcium concentration ([Ca2+]i) stimulated the [Na+]i-activated INa-Ca up to 1 microM [Ca2+]i, then inhibited the exchange current at very high [Ca2+]i (1 mM). 5. A step decrease in cytoplasmic pH from 7.2 to 6.4 (increase in [H+]i) produced a biphasic but monotonic decrease in INa-Ca. Alkalinization of cytoplasmic pH from 7.2 to 8.0 caused a large, biphasic increase in INa-Ca. 6. When INa-Ca was activated by a step increase in [Na+]i and [H+]i was simultaneously increased, the outward current rose to a peak and then declined to a low steady level. The peak current seen was always less than the maximum current produced by an identical elevation of [Na+]i at constant pHi. This reduction in peak outward current reflected a rapid 'primary' inhibition of the sodium-calcium exchange by protons. The decay of the sodium-calcium exchange current following the peak was slow and corresponded to the time course of the onset of a 'secondary' proton block. 7. Rapid primary inhibition of the sodium calcium exchanger could also be produced by cytoplasmic acidification in the absence of cytoplasmic sodium. The primary blockade was revealed when a subsequent increase in [Na+]i activated INa-Ca and a smaller peak outward current was observed. Secondary inhibition of the sodium-calcium exchanger was not, however, produced by cytoplasmic acidification in the absence of cytoplasmic sodium. Regardless of the duration of exposure to elevated [H+]i, the 'secondary' block by protons was still seen on activation of INa-Ca by increased [Na+]i as a gradual reduction of outward current amplitude. 8. Treatment of the sodium calcium exchanger with the proteolytic enzyme alpha-chymotrypsin largely removed its sensitivity to protons. 9. We conclude that the action of alpha-chymotrypsin on the monomeric sodium-calcium exchange protein is in part to remove a proton sensitive regulatory component(s) or render the regulation ineffective.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410704 TI - Silent period evoked by transcranial stimulation of the human cortex and cervicomedullary junction. AB - 1. The silent period evoked in the first dorsal interosseous (FDI) muscle after electrical and magnetic transcranial stimulation (TCS), electrical stimulation of the cervicomedullary junction and ulnar nerve stimulation was studied in ten healthy subjects. 2. With maximum-intensity shocks, the average duration of the silent period was 200 ms after electrical TCS, 300 ms after magnetic TCS, 43 ms after stimulation at the cervicomedullary junction and 100 ms after peripheral nerve stimulation. 3. The duration of the silent period, the amplitude of the motor-evoked potential, and the twitch force produced in the muscle were compared at increasing intensities of magnetic TCS. When the stimulus strength was increased from 30 to 70% of the stimulator output, the duration of the silent period lengthened as the amplitude of the motor potential and force of the muscle twitch increased. At 70 to 100% of the output, the amplitude of the motor potential and force of the muscle twitch saturated, whereas the duration of the silent period continued to increase. 4. Proximal arm muscle twitches induced by direct electrical stimulation of the biceps and extensor wrist muscles produced no inhibition of voluntary activity in the contracting FDI muscle. 5. The level of background activation had no effect on the duration of the silent period recorded in the FDI muscle after magnetic TCS. 6. Corticomotoneurone excitability after TCS was studied by means of a single magnetic conditioning shock and a test stimulus consisting either of one single magnetic shock or single and double electrical shocks (interstimulus interval 1.8 ms) in the relaxed muscle. A conditioning magnetic shock completely suppressed the response evoked by a second magnetic shock, reduced the size of the response evoked by a single electrical shock but did not affect the response evoked by double electrical shocks. Inhibition of the test magnetic shock was also present during muscle contraction. 7. Our findings indicate that the first 50 ms of the silent period after TCS are produced mainly by spinal mechanisms such as after-hyperpolarization and recurrent inhibition of the spinal motoneurones. If descending inhibitory fibres contribute, their contribution is small. Changes in proprioceptive input probably have a minor influence. From 50 ms onwards the silent period is produced mainly by cortical inhibitory mechanisms. PMID- 8410705 TI - Depression of tetanic force induced by loaded shortening of frog muscle fibres. AB - 1. Single fibres isolated from the anterior tibialis muscle of Rana temporaria were allowed to shorten against a high load during a 2.5-4.0 s fused tetanus (1-3 degrees C) and the maximum force produced at the short length was compared with that recorded during a fixed-end tetanus at the same overall fibre length. Changes in length of marked, consecutive segments (ca 0.5 mm in length) along the fibre were measured throughout the tetanus using a photoelectric recording system. 2. Loaded shortening (load ca 3/4 of maximum tetanic force) starting from approximately 2.55 microns sarcomere length and ending near slack fibre length depressed the tetanic force by 13 +/- 2% (mean +/- S.E.M., n = 10) and caused a marked redistribution of sarcomere length along the fibre. Unloaded shortening over the same range caused no force deficit and did not lead to increased dispersion of sarcomere length. 3. Loaded shortening below slack length produced less force depression and less non-uniformity of sarcomere length than did a corresponding intervention above slack length. 4. The force deficit after loaded shortening, both above and below slack fibre length, was positively correlated (P < 0.005) to the coefficient of variation of the sarcomere length along the fibre. 5. The decrease in active force after loaded shortening, and its relation to increased dispersion of sarcomere length along the fibre, could be simulated closely by a computer model in which the muscle fibre was assumed to consist of eleven discrete segments acting in series with a passive elastic element. 6. Experiments were performed in which the length of an individual segment of the intact muscle fibre was strictly controlled throughout a tetanus. Loaded shortening of such a 'length-clamped' segment caused no force depression during the subsequent isometric phase either above or below slack fibre length. 7. The results suggest strongly that force depression after loaded shortening of a single muscle fibre is attributable to non-uniform sarcomere behaviour along the fibre. The experimental evidence supports the view that: (i) the myosin cross bridges act as independent force generators; and (ii) their steady-state performance during a tetanus is unaffected by the preceding contractile activity. PMID- 8410706 TI - Polarity reversal of the optical rotation signals with change in direction of impulse conduction along the lobster nerve. AB - 1. The optical rotation signal of nerve associated with excitation was recorded from peripheral nerve taken from a walking leg of a spiny lobster and its properties were analysed. 2. The polarity of the optical rotation signal was reversed when the site of stimulation was changed with reference to the site of optical recording, so that the direction of impulse conduction was reversed, in most of the preparations. 3. Apart from the main response, which is associated with the conducted impulse, a pre-response was found to exist, which manifested itself on anodic stimulation, in a tetrodotoxin-treated nerve, or during the refractory period of the nerve, when the site of stimulation was close to the site of optical recording. The polarity of the pre-response was also reversed when the site of stimulation was changed with reference to the site of optical recording. 4. When the nerve was inclined from the horizontal level, so that the angle of incidence of light to the nerve was changed, the main response changed its amplitude and sometimes its polarity, whereas the pre-response remained practically unchanged. Thus the dependence on the angle of incidence was different between the pre-response and the main response. 5. It is suggested that the dependence of amplitude and polarity of the main response on the angle of incidence of light cannot be explained by the change in molecular axes of the membrane macromolecules, but can only be explained by their conformational change; and therefore the main response can be used as a monitor for the molecular conformation. PMID- 8410707 TI - Compartmentation of creatine kinase isoenzymes in myometrium of gravid guinea pig. AB - 1. This study was performed to investigate the possible presence and role of the creatine kinase (CK) system in the contraction and relaxation of skinned guinea pig uterus as well as the changes of the CK system during gestation. Experiments were performed on isolated longitudinal fibres of gravid and non-gravid myometrium. 2. Total CK activity increased from 74 +/- 11 to 196 +/- 39 IU (g wet wt)-1 during gestation. 3. The four isoenzymes of CK: muscle (MM), muscle-brain (MB), brain (BB) and mitochondrial (mt-CK) were found in myometrium. MM, MB and BB isoenzymes represented respectively 20.3 +/- 2.6, 10.3 +/- 4.4 and 72.7 +/- 2.2% of total activity. The distribution of isoenzymes did not significantly change with gestation, the contribution of mt-CK increasing from trace to 5% of total activity. 4. BB-CK was specifically bound to Triton X-100-skinned fibres with the non-gravid uterus containing 6.7 +/- 1.9 IU (g wet wt)-1 and the gravid uterus containing 44 +/- 13 IU (g wet wt)-1. 5. Active tension of Triton X-100 treated fibres increased from 6.06 +/- 0.68 to 19.3 +/- 1.9 mM mm-2 during gestation. 6. Submaximal tension (43.3 +/- 4.4% of maximal tension) can be developed in the absence of ATP and in the presence of 12 mM phosphocreatine (PCr) and 250 microM MgADP from endogenous CK in non-gravid uterine fibres while the gravid uterus was able to generate 65.4 +/- 3.9% of maximal tension via the CK system. 7. The endogenous CK system was able to relax the skinned fibres from high-tension rigor conditions by 47.3 +/- 4.2% of total relaxation in non-gravid fibres and 60.6 +/- 3.2% of total relaxation in gravid fibres. 8. Non-gravid and gravid uteri both contained mt-CK of 17.5 +/- 8.4 and 140 +/- 22 micrograms (g wet wt)-1 respectively as determined with antibodies against mt-CK. 9. Oxygen consumption was studied in fibres where the plasmalemma was solubilized with 50 micrograms ml-1 saponin. Maximal respiration was increased from 0.91 +/- 0.05 to 2.61 +/- 0.16 mumol oxygen min-1 (g dry wt)-1 in the gravid uterine fibres. However, creatine did not stimulate respiration in the uterine fibres treated with saponin. 10. It is concluded that the CK system undergoes qualitative as well as quantitative changes during gestation. BB-CK is specifically localized in the myofilaments and mt-CK is present in the uterine mitochondria.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410708 TI - Modification of carotid chemoreceptor-induced changes in renal haemodynamics and function by carotid baroreflex in dogs. AB - 1. Mongrel dogs were anaesthetized with thiopental sodium and chloralose and artificially ventilated. The carotid sinus regions were vascularly isolated and perfused either with arterial or mixed (arterial and venous) blood (PO2, 44.2 +/- 3.3 mmHg, mean +/- S.E.M.) to stimulate the chemoreceptors. Cervical vagosympathetic trunks were ligated and atenolol (2 mg kg-1, I.V.) was given in all dogs and gallamine triethiodide (3 mg kg-1 h-1, I.V.) was given in two dogs. Renal blood flow was measured by an electromagnetic flowmeter, glomerular filtration rate by creatinine clearance, sodium excretion by flame photometry and solute excretion by osmometry. The viability of the preparations was tested by recording total vascular capacitance responses to stimulation of carotid baro- and chemoreceptors. 2. In eleven tests in seven dogs at a constant aortic pressure of 88.9 +/- 2.6 mmHg stimulation of carotid chemoreceptors at a high carotid sinus pressure (194.0 +/- 3.6 mmHg) resulted in significant increases in urine flow of 22.8 +/- 3.0%, urinary sodium excretion of 30.7 +/- 5.2%, fractional sodium excretion of 35.3 +/- 18.6%, osmolar excretion of 17.5 +/- 4.1% and a decrease in free water clearance of 30.8 +/- 3.1% without significant changes in urinary sodium concentration, renal blood flow, glomerular filtration rate, and filtration fraction. 3. In seventeen tests in these seven dogs at a constant aortic pressure of 94.0 +/- 2.2 mmHg, stimulation of carotid chemoreceptor at a low carotid sinus pressure (72.0 +/- 1.3 mmHg) resulted in significant decreases in renal blood flow of 10.6 +/- 2.5%, glomerular filtration rate of 19.6 +/- 6.8%, filtration fraction of 13.2 +/- 5.5%, urine flow of 23.4 +/- 4.1%, urinary sodium concentration of 20.3 +/- 4.1%, urinary sodium excretion of 38.5 +/- 4.6%, fractional sodium excretion of 20.2 +/- 7.7%, osmolar excretion of 23.9 +/- 4.0% and an increase in free water clearance of 23.1 +/- 2.5%. 4. The results show that moderate stimulation of carotid chemoreceptors at a low carotid sinus pressure, when the activity in renal nerves is high and blood volume is low, can produce significant reflex decreases in renal haemodynamic and functional variables. However, at a high carotid sinus pressure when the renal sympathetic activity is low and blood volume is high, carotid chemoreceptor stimulation produces diuresis and natriuresis but no change in renal haemodynamics. PMID- 8410709 TI - The influence of intracellular pH on contraction, relaxation and [Ca2+]i in intact single fibres from mouse muscle. AB - 1. The effects of intracellular pH (pHi) on myoplasmic free calcium concentration ([Ca2+]i) and contractile performance were studied in intact single fibres dissected from mouse skeletal muscle. Indo-1 was used to measure [Ca2+]i and pHi was altered by changing perfusate CO2. 2. Tetanic tension was decreased at acidic pHi and increased at alkaline pHi whereas the rate of mechanical relaxation was showed at both acidic and alkaline pHi. Resting and tetanic [Ca2+]i were increased at acidic pHi and decreased at alkaline pHi while the final rate of decline of [Ca2+]i after a tetanus was markedly slowed at acid pHi but only marginally accelerated at alkaline pHi. 3. Steady-state [Ca2+]i-tension curves were constructed from measurements of tetani at different stimulus frequencies. The curves at acid pHi showed a reduced maximum Ca(2+)-activated tension and a reduced Ca2+ sensitivity, and curves at alkaline pHi showed the opposite changes. 4. Two methods were devised to determine the contribution of [Ca2+]i to the rate of relaxation. In one method the instantaneous tension was plotted as a function of instantaneous [Ca2+]i throughout a tetanus and compared with the steady-state [Ca2+]i-tension relation. In a second method the [Ca2+]i signal during a tetanus was converted to a Ca(2+)-derived tension record by means of the steady-state [Ca2+]i-tension relation and this Ca(2+)-derived tension was then compared to the true tension. 5. The sarcoplasmic reticulum (SR) pump function was analysed by plotting -d[Ca2+]i/dt against [Ca2+]i during the final slow decline of [Ca2+]i after a tetanus. This analysis shows that the Ca2+ uptake by the SR is a third- or fourth-power function of [Ca2+]i and that acidosis substantially slows the rate of SR Ca2+ pumping. 6. In conclusion, the slowing of relaxation at acidic pHi could be attributed to a slowing of cross-bridge detachment rather than the observed slowing of the rate of uptake of Ca2+. Conversely the slowing of relaxation in alkaline pHi could be attributed to the increase of Ca2+ sensitivity of the myofibrillar proteins. PMID- 8410710 TI - Ion permeation through the L-type Ca2+ channel in rat phaeochromocytoma cells: two sets of ion binding sites in the pore. AB - 1. Both inward and outward unitary Li+ currents through the L-type Ca2+ channel and discrete block of such currents by either internal or external Ca2+ are recorded. Detailed kinetic analyses are obtained for all of the four experimental configurations (internal or external Ca2+ block of either inward or outward Li+ currents). 2. No matter from which side the blocking Ca2+ ion comes, the exit (unblocking) rates are always the same at the same potential in the same direction of Li+ current flow. This indicates that the high-affinity Ca2+ binding site (the blocking site) is in the pore, and internal and external Ca2+ both go to the same site to produce the block. In other words, there can only be one high affinity Ca2+ binding site or one set of such sites (sites separated by insignificant barriers) in the pore. Furthermore, the direction of exit of the blocking Ca2+ ion is always with, not against, the Li+ current flow. This suggests ion-ion interaction (the 'long pore effect') in the high-affinity sites. Therefore there must be more than one high-affinity site in the pore. Overall it is concluded that the pore must contain a set of high-affinity Ca2+ binding sites separated by insignificant energy barriers. 3. The voltage dependence of the off- (exit) rates is very similar in amplitude for all the four experimental configurations (e-fold change per approximately 25 mV depolarization or hyperpolarization). This strong voltage dependence in every configuration cannot be explained by any Ca2+ energy profile alone and must include a certain contribution from Li+. The mechanism underlying such a contribution seems to reside in the enhancement effect of Li+ on the exist of Ca2+. 4. The on-rates (blocking rates) for external Ca2+ are always fast no matter whether the Li+ currents are outward or inward. In certain cases the rates even approach the diffusion-controlled limit (approximately 10(9) M-1 S-1). This suggests that the high-affinity sites are very easily accessible from the outside, and probably there is no other ionic site located between the external pore mouth and the high affinity sites. 5. The on-rates for internal Ca2+ are fast and voltage independent in outward Li+ currents, but are very slow and strongly voltage dependent in inward Li+ currents.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410711 TI - Characterization of the high-affinity Ca2+ binding sites in the L-type Ca2+ channel pore in rat phaeochromocytoma cells. AB - 1. Two major types of ion-ion interaction are demonstrated in the pore of the L type Ca2+ channel, the 'lock-in' and the 'enhancement' effect. The former denotes that the ion at a certain site in the pore cannot move if the neighbouring site is occupied by the other ion. The latter denotes that the ion occupying a certain site may facilitate the exist of the other ion in the neighbouring site. 2. With inward currents carried by 300 mM external Li+, the exit rates of the blocking Ca2+ ion are decreased by approximately 4 times when the internal Li+ concentration is increased from 55 to 300 mM. With outward currents carried by 300 mM internal Li+, the exist rates of the blocking Ca2+ ion are decreased by approximately 2.5 times when the external Li+ concentration is increased from 55 to 300 mM. These findings demonstrate the 'lock-in' effect. 3. When inward currents are carried in Li+ in the cell-attached configuration, the on-rates of the external Ca2+ are decreased in a rectangular hyperbolic fashion with increasing external Li+ concentration (apparent dissociation constant (Kd) approximately 75 mM in activity), suggesting competition between Ca2+ and Li+ for a certain site. On the other hand, the off-rates of the blocking Ca2+ ion are increased linearly with increasing external Li+ concentration between 75 and 850 mM, and the line extrapolates to the zero point, indicating that Ca2+ exist is negligible at zero Li+. This finding demonstrates not only the existence of the enhancement effect in the channel, but also the indispensability of such an effect for Ca2+ to exit the pore. Moreover, a linear relationship up to 850 mM Li+ suggests that the affinity of Li+ to the enhancement sites is very low (apparent Kd very high) when a Ca2+ ion is present in the neighbouring site. 4. The unitary conductances of inward Ba2+ currents in the cell-attached configuration are increased with increasing Ba2+ concentration. The apparent Kd obtained from a rectangular hyperbolic fit to the data is approximately 6 mM Ba2+ (in activity). When inward Ba2+ currents are blocked by Cd2+, the on-rates of Cd2+ are decreased with increasing Ba2+ concentration also in a rectangular hyperbolic fashion, and the apparent Kd is approximately 5.5 mM. The similar Kd from these two different experiments suggests the high-affinity set can accommodate no more than two Ba2+ ions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410712 TI - Block of the L-type Ca2+ channel pore by external and internal Mg2+ in rat phaeochromocytoma cells. AB - 1. Three to eight micromolar external Mg2+ produces discrete block of the unitary inward currents through the L-type Ca2+ channel carried by 300 mM external Li+. Like the Ca2+ block, increasing Li+ concentration decreases the Mg2+ on-rate and increases the Mg2+ off-rate. 2. These kinetic changes are saturating and the apparent dissociation constant (Kd) for the on-rates in 75 mM Li+ (in activity), the same as that in the case of Ca2+ block. This suggests that Mg2+ and Ca2+ produce the discrete block at the same site. The apparent Kd for the off-rates is 300 mM, much smaller than that in the case of Ca2+ block. This indicates that Mg2+ exerts much less repulsion on the Li+ ion in the neighbouring (enhancement) site than Ca2+, although Mg2+ and Ca2+ both have two charges. The theoretical fits to the off-rates also suggest that Mg2+ can exit the blocking sites at a rate of several hundred per second in the absence of any enhancement effect. 3. Seventeen to forty-eight micromolar internal Mg2+ produces discrete block of the outward unitary currents carried by 300 mM internal Li+. The off-rates are in general approximately 20 times faster as compared to the Mg2+ off-rates in the inward currents. This finding suggests that Mg2+ in the high-affinity sites can much more easily exit to the outside than to the inside, implying significantly higher energy barriers on the inner side of the high-affinity sites for Mg2+. 4. At least 5-10 mM internal Mg2+ is needed to produce discrete block of the inward unitary currents carried by 215 mM external Na+. The off-rates in such experiments are generally the same as those in the case of external Mg2+ block of inward currents. This suggests that internal and external Mg2+ both reach the same site, namely the high-affinity Ca2+ binding sites in the pore, to produce the discrete block. 5. Other than discrete block, 5-10 mM internal Mg2+ also decreases the size of the inward unitary current. This is most probably due to a fast block at the more internally located low-affinity sites in the pore. The fractional decrease of the currents is voltage dependent and can be fitted by a rectangular hyperbola to calculate the apparent Kd, which increases e-fold per 45 mV hyperpolarization, indicating an electrical distance of 0.3 between the low affinity sites and the internal pore mouth.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410713 TI - Modification of the adenosine 5'-triphosphate-sensitive K+ channel by trypsin in guinea-pig ventricular myocytes. AB - 1. The adenosine 5'-triphosphate (ATP)-sensitive K+ channel current was recorded in guinea-pig ventricular myocytes using the patch clamp technique with inside out patch configuration. Modification of the channel activity by intracellular application of an endoprotease trypsin was studied, and was related to a possible model of regulation of this channel. 2. Maximal ATP-sensitive K+ channel activity was observed immediately upon formation of inside-out patches in the ATP-free internal solution, thereafter activity declined both spontaneously and gradually with time; a phenomenon known as rundown. When trypsin (1 mg/ml) was applied to the intracellular side of the membrane upon formation of inside-out patches, spontaneous run-down did not occur, and this trypsin action was irreversible. Neither trypsin (1 mg/ml) applied with trypsin inhibitor (0.25 mg/ml) nor heat denatured trypsin (1 mg/ml) could mimic this effect. When trypsin was applied to the patches after run-down, channels were reactivated at approximately 13 min. 3. Treatment with trypsin did not affect unitary current amplitude, channel gating kinetics, or sensitivity to intracellular ATP. 4. Intracellularly applied Ca2+ induced run-down of channel activity in a dose-dependent manner. In membrane patches that were treated with trypsin (1 mg/ml) for 20 min, intracellularly applied Ca2+ up to 1 mM did not induce run-down of channel activity. 5. Intracellular application of an exopeptidase, carboxypeptidase A (1 mg/ml), but not Leu-aminopeptidase (0.5 mg/ml), prevented spontaneous or Ca(2+)-induced run down of channel activity. 6. As postulated for several other channels, such as Na+ and Ca2+ channels, there may be a possible 'chemical gate' that is responsible for run-down of this channel activity. Application of trypsin might somehow modify this 'chemical gate', resulting in prevention of spontaneous or Ca(2+)-induced run-down. This target site for trypsin may be situated on the carboxy-terminus of the channel proteins, or of associated regulatory units. Because ATP sensitivity remained intact after trypsin treatment, the trypsin selective site for channel inhibition is not related physically to the ATP binding site. PMID- 8410714 TI - Nifedipine- and omega-conotoxin-sensitive Ca2+ conductances in guinea-pig substantia nigra pars compacta neurones. AB - 1. The membrane properties of substantia nigra pars compacta (SNc) neurones were recorded in guinea-pig in vitro brain slices. 2. In the presence of tetrodotoxin (TTX) a Ca(2+)-dependent slow oscillatory potential (SOP) was generated. Application of 0.5-20 microM nifedipine abolished both spontaneous and evoked SOPs. A tetraethylammonium chloride (TEA)-promoted high-threshold Ca2+ spike (HTS) was little affected by nifedipine. On the other hand, omega-conotoxin applied either locally or via the perfusion medium (1-10 microM) blocked a part of the HTS, but it did not alter the SOP. 3. In normal medium nifedipine blocked the spontaneous discharge, decreased the interspike interval (ISI) recorded during depolarizing current injections and selectively reduced the slow component of the spike after-hyperpolarization (AHP). omega-Conotoxin decreased both the rising and falling slopes of the normal action potential, reduced the peak amplitude of the spike AHP, and, in some of the neurones, reduced the ISI during depolarization. The Na+ spikes recorded in Ca(2+)-free medium were not altered by omega-conotoxin. 4. The SOP was not blocked by octanol (100-200 microM), amiloride (100-250 microM), or Ni2+ (100-300 microM). However, at 500 microM Ni2+ attenuated the SOP. 5. Application of apamin (0.5-2.0 microM) induced irregular firing or bursting, abolished the slow component of the spike AHP and reduced its peak amplitude. In the presence of TTX and apamin long-duration plateau potentials occurred, which were subsequently blocked by nifedipine. 6. In Ca(2+) free, Co(2+)-containing medium TTX-sensitive spikes and voltage plateaux were generated by depolarizing current pulses. It is suggested that a persistent Na+ conductance underlies the plateaux, which may be co-activated during the SOP. 7. The results suggest that the Ca2+ currents underlying the SOP and the HTS are different and that they activate at least two Ca(2+)-dependent K+ conductances. These conductances play major roles in the maintenance of spontaneous discharge and in control of membrane excitability. PMID- 8410715 TI - cAMP-activated chloride currents in amphibian retinal pigment epithelial cells. AB - 1. The effect of cAMP on whole-cell currents in isolated retinal pigment epithelial (RPE) cells of the bullfrog and marine toad was investigated by means of the perforated patch clamp technique. 2. Superfusing cells with either cAMP or forskolin led to the development of a time-independent current that had a linear current-voltage (I-V) relationship. The reversal potential of (Vrev) of the cAMP activated current was unaffected by the removal of either Na+ or HCO3- from the external and internal solutions or by the addition of extracellular barium, but it was near the Cl- equilibrium potential (ECl) over a wide range of extracellular Cl- concentrations, suggesting the presence of a Cl(-)-selective channel. 3. The anion permeability sequence of the cAMP-activated conductance calculated from biionic reversal potentials was NO3- = I- > Br- > Cl- >> HCO3- > methanesulphonate. 4. The conductance was blocked by a variety of Cl- transport inhibitors, including 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS), 4,4'-dinitro-2,2'- stilbene disulphonic acid (DNDS), frusemide, N phenylanthranilic acid (DPC) and niflumic acid. 5. The present study demonstrates that cAMP activates a Cl(-)-selective channel that most probably resides in the basolateral membrane. PMID- 8410716 TI - Effects of temperature on calcium current of bullfrog sympathetic neurons. AB - 1. The temperature dependence of whole-cell calcium current was studied in bullfrog sympathetic neurons. 2. The factor by which the peak calcium current increased upon a 10 degrees C increase in temperature (Q10) was 1.6 +/- 0.1 (mean +/- S.E.M., n = 9), for a change from 20 to 30 degrees C. 3. Activation and deactivation were more rapid at a higher temperature. The Q10 was approximately 7 in the middle of the voltage range (near -10 mV), where the kinetics were slowest. Time constants were less temperature dependent at more positive or negative voltages (Q10 approximately 2 at -70 mV). 4. Near -10 mV, activation and deactivation were associated with a large enthalpy and a large positive entropy change. Deactivation at -70 mV reflected a smaller enthalpy change, and almost no change in entropy. 5. Activation is only slightly more temperature dependent than deactivation, when both are measured at the same voltage. 6. The peak current shifts slightly (approximately 5 mV) to more negative voltages upon a change from 20 to 30 degrees C. 7. Inactivation has a Q10 of approximately 2 at -10 mV. 8. Changes in the kinetics of activation or inactivation could be observed during recording at a constant temperature. These changes were generally small, especially for activation kinetics, and could be distinguished from temperature dependent changes. 9. The changes in entropy and Q10 with voltage suggest that the rate-limiting steps for activation and deactivation are different at extreme voltages vs. the middle of the voltage range. PMID- 8410717 TI - Kinetic analysis of two types of Na+ channels in rat dorsal root ganglia. AB - 1. The gating properties of two types of Na+ channels were studied in neurones isolated from rat dorsal root ganglia using the whole cell variation of the patch electrode voltage-clamp technique. 2. Two types of Na+ currents (INa) were identified on the basis of their sensitivity to tetrodotoxin (TTX). One type was insensitive to TTX (up to 0.1 mM), while the other type was blocked by 1 nM of TTX. Whereas they were both insensitive to 50 microM Cd2+, a high concentration (2 mM) of Co2+ selectively inhibited the TTX-insensitive type. 3. The activation thresholds were about -60 and -40 mV for the TTX-sensitive and the TTX insensitive INa, respectively. Activation of the TTX-sensitive INa developed with a sigmoidal time course which was described by m3 kinetics, whereas the activation of the TTX-insensitive INa was described by a single exponential function. A deactivation process, as measured by the tail current upon repolarization, followed an exponential decay in either type of INa. 4. The rate constant of activation indicated that under comparable membrane potential conditions, the TTX-insensitive channels open 4-5 times slower than the TTX sensitive ones upon depolarization. Likewise, the rate constant of inactivation indicated that the TTX-insensitive channels inactivate 3-7 times more slowly than the TTX-sensitive ones upon repolarization. 5. The steady-state activation curve for the TTX-insensitive INa was shifted about 20 mV in the positive direction from that for the TTX-sensitive INa. 6. The steady-state inactivation curve for the TTX-insensitive INa as obtained with a 0.5 s prepulse was shifted about 26 mV in the positive direction from that for the TTX-sensitive INa, indicating a greater availability for the TTX-insensitive INa in depolarized membrane. However, on increasing the duration of prepulse, the inactivation curve for the TTX-insensitive INa, but not for the TTX-sensitive INa, shifted in the negative direction due to an extremely slow inactivation process in the TTX-insensitive INa. Consequently, an overlap between the activation and inactivation curves which causes a steady influx of Na+ (window current) became progressively reduce. 7. The time course of INa decay was best described by a single exponential process in either the TTX-sensitive or TTX-insensitive INa, whereas the development of inactivation and the recovery from inactivation, which were measured by a conventional double-pulse protocol, followed a second order process in either channel type.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8410718 TI - A correlation between quantal content and decay time of endplate currents in frog muscles with intact cholinesterase. AB - 1. The relationship between quantal content and prolongation of endplate currents (EPC) was studied in the frog sartorius with intact synaptic acetylcholinesterase. 2. The prolongation of EPC was more pronounced in endplates with a higher quantal content both before and after potentiation of quantal release by 4-aminopyridine (4-AP). When the quantal content of EPC was lowered, either by high Mg2+ or repetitive stimulation, the EPC decay constant was reduced. 3. A certain critical value of about 120 quanta per nerve impulse was found, at which point the decay of EPC remained constant even through the quantal content was reduced further. 4. The reduction in both density and number of postsynaptic receptors, produced by alpha-bungarotoxin and (+)-tubocurarine led to a profound reduction in EPC decay during the progressive fall in EPC amplitude in both 4-AP-treated and -untreated endplates. Both drugs are known to produce a shortening of EPC in anti-cholinesterase (anti-ChE)-treated muscles, due to a decrease in receptor density and less frequent repetitive binding of ACh. 5. It is assumed that the prolongation of multiquantal EPC is caused by an increased ACh concentration near the receptors, which may provide the opportunity for repetitive binding even with full cholinesterase activity. The critical quantum content of about 120 might be the number of quanta at which the probability of multiple release at single active zones is increased above zero. PMID- 8410719 TI - The LP/VN role in combating hypertension: "The Silent Killer". PMID- 8410720 TI - Chronic fatigue syndrome: what are the facts? PMID- 8410721 TI - How humor helps nurses manage job stress. PMID- 8410723 TI - Myth vs. fact about ulcer disease. PMID- 8410722 TI - Eating disorders: anorexia nervosa & bulimia. PMID- 8410724 TI - Esthetic recontouring after surgical and orthodontic treatment of an impacted and developmentally impaired permanent incisor: a case report. PMID- 8410725 TI - Transmolar pin and magnetic carrier for midfacial reconstruction: a clinical report. AB - With previous prosthetic restorations of bilateral maxillary resections, tissue erosion and bleeding on the cephalic parts of the cavity have been caused by movement and pressure from the prosthesis. Retention of a prosthesis is a major determinant in its successful use. Physical and psychologic stresses are common when one is unable to use a prosthesis comfortably. The use of a musculocutaneous flap and an obturator prosthesis had provided near-immediate reconstruction of a massive midfacial defect. This method of reconstruction resulted in soft tissue replacement for appearance and functional rehabilitation of the upper lip while maintaining sound oncologic principles. PMID- 8410726 TI - Localized three-body wear of six types of composite resin veneering materials. AB - The wear resistance of six types of composite resin veneering materials was determined in vitro by use of a three-body wear-testing device. This investigation examined the wear characteristics of the veneering composite resins in heavy occlusal contact areas. After repeated cyclic loading, wear values were measured with a profilometer, and surface textures were evaluated with a scanning electron microscope. All of the composite resin veneering materials were significantly more wear resistant than unfilled acrylic resin; but the composite resin materials demonstrated rougher surfaces than unfilled resin. Three of the composite resins exhibited fractures in the contact areas. PMID- 8410727 TI - The effect of unfilled resin dilution on composite resin hardness and abrasion resistance. AB - Diluting composite materials with unfilled resin to facilitate clinical application during direct lamination will result in an alteration of filler loading with possible changes of physical properties. This study compared the surface hardness and abrasion resistance of a visible light-cured microfilled and hybrid resin with and without dilution. Samples were prepared, cured, and stored in distilled water at 37 degrees C for 24 hours before testing. Knoop hardness values were obtained from the upper surface of 10 samples in three dilution and control groups of each material. Three samples of the dilution and control groups were similarly prepared and stored for toothbrush abrasion testing. Data from the hardness and abrasion tests were subjected to analysis of variance and Tukey's test at a 0.05 level of significance. Results showed that dilution had a significant effect on surface hardness of the materials tested, although the diluted materials were not significantly different from the controls. Abrasion tests showed visible evidence of abrasion on all samples, but dilution showed no significant effect on abrasion resistance of the materials tested. PMID- 8410728 TI - Increase of in vitro curing depth of class II composite resin restorations. AB - Simulated class II cavity preparations were made in black Plexiglas acrylic, and various posterior composite resins were placed in the preparations and cured with and without a transparent cone-shaped "Light-tip" device. The transparent Light tip device improved the curing depth approximately 2.2 mm, and this ability to illuminate the deeper portions of the restoration increases the potential to improve the curing of composite resin. PMID- 8410729 TI - Effect of acid etching on the flexural strength of a feldspathic porcelain and a castable glass ceramic. AB - This project studied the effect of altering surface topography by chemical etching on the strength of a feldspathic porcelain and castable glass ceramic. Fifty specimens of each ceramic material were subjected to five different etch times (in groups of 10). A silane coupling agent and composite resin cement were applied. Specimens were then subjected to a three-point flexural strength test. Representative specimens were examined under scanning electron microscope to elucidate more information on the effect and the depth of etch. There was no significant difference in the mean flexural strengths between the etched and nonetched groups and no significant difference between the different etching times for either material. Photomicrographs revealed dissimilar etch depths and selective dissolution of the phases. It appears that chemical etching can improve the retention of ceramic laminate veneers without significant loss of strength. PMID- 8410730 TI - Comparison of the retentive properties of two hollow-tube post systems to those of a solid post design. AB - The retention of a post in a root is critical for the successful restoration of an endodontically treated tooth. This study compared the retention of two hollow tube post designs (Access post and World post) cemented with zinc phosphate cement or a titanium-reinforced composite resin cement (Flexi-Flow cement with titanium) with that of the control Para-Post system, a solid post design cemented with zinc phosphate cement. The experiment was divided into 11 groups of 10 samples. Retentive values were measured with a universal testing machine and the data were recorded in pounds. A one-way analysis of variance was computed and included Duncan's multiple comparisons test, Tukey's Student range test, and Dunnett's test. Duncan's multiple comparison test and Tukey's test were inconclusive. Only Dunnett's test revealed a significant difference between group 5, Access post, grooved dentin with Flexi-Flow cement with titanium (mean of 89.7 lb), and the control, group 11, Para-Post dowel cemented with zinc phosphate cement (mean of 61.3 lb). All other groups exhibited similar results with retentive values that ranged from 62 to 76 lb. PMID- 8410731 TI - Improving the accuracy of a new articulator. AB - The indications and features of the new Denar Combi articulator are reviewed. The Combi articulator was designed primarily as a set path instrument, but can also be used as a semiadjustable or fully adjustable stereographic articular. The articulator can be programmed for any of these modes, but the Combi model is limited when used as a stereographic instrument according to the manufacturer's instructions, with the instrument-precipitated geometric errors. These errors were identified mathematically, and an accurate technique for the instrument is described. PMID- 8410732 TI - Maxillary distal-extension removable partial denture abutments with reduced periodontal support. AB - An in vitro study that used photoelastic models compared stress distribution characteristics of three maxillary, bilateral, distal-extension removable partial denture designs when the abutments were subjected to a progressive loss of periodontal support. One design used I-bar retention, a second design used a semiprecision, spring-loaded plunger attachment, and a third design used the ERA attachment. Both attachment designs were tested with and without splinted abutments. The ERA design was also tested with and without supporting rests and included light and heavy retention elements. Periodontal support loss up to 35%, a 60/40 crown-to-root ratio, resulted in increased stress concentrations. The ERA design with supporting rests, light retention elements, and splinted abutments compared very favorably with the I-bar retained design on nonsplinted abutments. PMID- 8410733 TI - A clinical evaluation of conical crown retained dentures. AB - In an uncontrolled retrospective recall study of 57 patients treated with conical crown retained dentures, 60 restorations (37 in the maxillae and 23 in the mandible jaw) with a mean wearing time of 30.1 months (range 4 to 76) were evaluated. Of the 248 abutments, eight (3.2%) had been lost. Clinically healthy mucosa was seen in 35 jaws. The marginal fit of the copings was judged to be good. No caries or new restorations were observed in 44 patients. Thirteen patients had 19 surfaces with new restorations and 20 surfaces with caries lesions. Of these 39 surfaces, 38 were located subgingivally. PMID- 8410734 TI - Diseases, medication, and postinsertion visits in complete denture wearers. AB - The records of 268 patients were used to assess the effects of five disease/drug complexes on the number of postinsertion visits in complete denture wearers. The data were analyzed with SAS and BMDP computer packages. The results showed a statistically significant increase in the number of postinsertion visits in patients who had central nervous system or psychiatric disorders. Practitioners are alerted to consider the ergonomic implications at the outset of treatment. PMID- 8410735 TI - A pilot study of exposure of the smear layer to tannic acid solutions. AB - This study examined the effects of tannic acid on smear layer removal. Class V cavities were prepared on the buccal surfaces of extracted human molars with a No. 1557 high-speed carbide bur under water cooling for cutting and a No. 701 low speed steel bur for finishing. They were then washed with water followed by an application of 2%, 5%, 10%, 15%, 20%, or 25% tannic acid solution in water for 15, 30, or 60 seconds. The axial wall dentin surfaces were examined with scanning electron microscopy. This examination showed that an application of 2% tannic acid for 60 seconds or 5% tannic acid for 15, 30, or 60 seconds effectively removed the smear layer, leaving the dentinal tubules occluded. The dentinal surfaces treated with 40% phosphoric acid for 15 seconds showed many exposed dentinal tubules with flared orifices. PMID- 8410736 TI - Identification of occlusal interferences on gold castings. PMID- 8410737 TI - A simple method for inspection of porosity in titanium castings. PMID- 8410738 TI - Adjustable pencil holder for a cerebral palsy patient. PMID- 8410739 TI - An official glossary is our best guide to effective communication. PMID- 8410740 TI - Organization of the larval pre-ecdysis motor pattern in the tobacco hornworm, Manduca sexta. AB - The tobacco hornworm, Manduca sexta, undergoes several larval molts before transforming into a pupa and then an adult moth. Each molt culminates in ecdysis, when the old cuticle is shed. Prior to each larval ecdysis, the old cuticle is loosened by pre-ecdysis behavior, which consists of rhythmic compressions that are synchronous along the abdomen and on both body sides, and rhythmic retractions of the abdominal prolegs. Both pre-ecdysis and ecdysis behaviors are triggered by a peptide, eclosion hormone. The aim of the present study was to investigate the neural circuitry underlying larval pre-ecdysis behavior. The pre ecdysis motor pattern was recorded in isolated nerve cords from eclosion hormone treated larvae, and the effects of connective transections and ionic manipulations were tested. Our results suggest that the larval pre-ecdysis compression motor pattern is coordinated and maintained by interneurons in the terminal abdominal ganglion that ascend the nerve cord without chemical synaptic relays; these interneurons make bilateral, probably monosynaptic, excitatory connections with identified pre-ecdysis motor neurons throughout the abdominal nerve cord. This model of the organization of the larval pre-ecdysis motor pattern should facilitate identification of the relevant interneurons, allowing future investigation of the neural basis of the developmental weakening of the pre-ecdysis motor pattern that accompanies the larval-pupal transformation. PMID- 8410741 TI - Discriminating parts from the whole: determinants of odor mixture perception in squirrel monkeys, Saimiri sciureus. AB - In a task designed to simulate olfactory-guided foraging, the ability of squirrel monkeys to discriminate an artificial 12-component odorant from 3-, 6-, 9- or 11 component submixtures was investigated. A combination of factors was found to contribute to the animals' performance: 1. Discriminability generally decreased as the number of components in the submixture increased. 2. Submixtures did not contribute equally to mixture perception, and one component in particular (cineole) disproportionately influenced stimulus discriminability. 3. Interactive effects between submixtures resulted in marked deviations from the general pattern of discriminability. 4. Changes in the relative concentration of submixtures could also influence discriminability. 5. Finally, individual differences in responsiveness to particular stimuli were apparent. These findings demonstrate that the interaction between components in odor mixtures can be complex and that seemingly small changes in composition can strongly affect perception and thus potential signal function. It is therefore suggested that in future investigations of squirrel monkey semiochemistry, the method of systematically varying submixtures may be particularly useful in defining the contribution of components to a signal. PMID- 8410742 TI - The effect of exercise on depression, anxiety and other mood states: a review. AB - This paper addresses the current literature related to investigations of the link between exercise treatments and depression, anxiety and other mood states. Results from these investigations are supportive of the anti-depressant, anti anxiety and mood enhancing effects of exercise programs. There were considered to be, however, a number of potential methodological problems in many of the research studies; the nature of these were considered. Finally, some possible directions for future research are outlined. PMID- 8410743 TI - Somatization misattributed to non-pathological vaginal discharge. AB - Lay explanations and attributional patterns about somatization related to vaginal discharge, a folk illness characterized by undue concern about passing (normal) vaginal discharge and believing that their symptoms are caused by this, were surveyed systematically in 200 consecutive women patients with predominant somatic complaints and 138 normal healthy women. Misattribution was reported 3.5 times more often by women with somatic complaints as compared to the normal women. Sixteen women were absolutely certain that their discharge was totally responsible for their symptoms and illness. Thus, somatization related to vaginal discharge seems to be a frequent undetected problem in women with somatic symptoms. PMID- 8410744 TI - Self-perceived fatigue and cortisol secretion in a community sample. AB - The possible relation of cortisol secretion to self-perceived fatigue was examined in an adult community sample. Serum cortisol levels measured after the dexamethasone suppression test (DST) did not predict fatigue whereas depression assessed by the Beck Depression Inventory (BDI) and current medical treatment were significant and independent predictors. Confounding variables such as age, gender, body mass and social status were not predictive of fatigue. Of putative hypothalamic-pituitary-adrenal abnormalities hypersecretion of cortisol does not typify subjects with self-perceived fatigue. PMID- 8410745 TI - Pain sensitivity in recovered anorexics, restrained and unrestrained eaters. AB - The heat pain threshold and local skin temperature were assessed in 23 former anorexic in-patients with an 'intermediate' (N = 9) or 'good' outcome (N = 14) and in 21 restrained and 20 unrestrained eaters. All subjects were female. The group means of the pain thresholds did not differ significantly from each other, suggesting that the homogeneous increase in pain thresholds we had previously observed in acutely ill eating disorder patients is state dependent. However, a sizeable percentage of the restrained eaters (29%) had pain thresholds clearly above the normal range. Thus it may well be that restrained eating carries a risk of reducing pain sensitivity. Pain threshold and skin temperature correlated significantly (r = -0.63) only in the group of patients with an intermediate outcome, a finding resembling that obtained in acute anorexics. This suggests that peripheral thermoregulation and pain sensitivity are linked in the acute and moderately improved phases of anorexia nervosa. PMID- 8410746 TI - Gender and cardiovascular responses: what is the role of hostility? AB - Two experiments are reported investigating hostility group by gender effects on heart rate and blood pressure. In experiment 1, 58 males and 59 females were tested with a protocol that included baseline, reaction time with and without harassment, and recovery. In experiment 2, 55 males and 50 females participated in a procedure of baseline, Stroop color--word conflict, anger recall interview, and recovery. Based on Buss-Durkee subscales, genders were divided into high and low neurotic hostility and expressed hostility groups. High expressed hostility was associated with greater diastolic reactivity to all tasks in males, but not in females. Low expressed hostility was associated with greater heart rate and systolic reactivity, specifically during the anger recall interview. High neurotic hostility in males and females was associated with greater diastolic reactivity to the anger task only, while low Neurotic hostility males had higher resting systolic pressures. These results suggest that outward expression of hostility in men may increase their coronary heart disease risk; however, results for both men and women support an association between cardiovascular reactivity and a lack of match between self-reported anger management style and task demands. PMID- 8410747 TI - Psychological morbidity in women referred for treatment of hirsutism. AB - Sixty-nine pre-menopausal hirsute women were studied to determine the prevalence of psychological morbidity, and to identify factors associated with psychiatric symptoms. Measures used included: GHQ and POMS to assess psychological morbidity; self-esteem; personality (EPQ); and social adjustment, amongst other. About a third of subjects were found to be GHQ cases, one third had never discussed their hirsutism before, and two thirds avoided some social situations. Psychological morbidity was associated with poor social adjustment, higher levels of neuroticism and introversion, and avoidance of some social situations, but not with the severity of hirsutism or testosterone levels. The results suggest that a proportion of women with hirsutism experiences psychological and social difficulties, and that factors other than their dermatological status play an important part in their psychological difficulties. PMID- 8410748 TI - Effects of some psychosocial variables on different disease manifestations in 112 cadets: a longitudinal study. AB - One-hundred and twelve cadets attending the 141st training course for reserve officers of the 'Scuola Transmission' of the Italian Army were administered, 10 days after incorporation, a battery of personality tests and measures of stressful events in the preceding year. Test scores were considered both individually and grouped into factors. During the 6 months of the course all disease episodes for each cadet were recorded. Total episodes infections and traumas were considered. A significantly higher number of total episodes and, specifically, of infections was present in subjects reporting a higher number of stressful events, in interaction with attitudes towards parent figures, hardiness, loneliness, and an alienation factor. A younger age of subjects also appeared predictive of a higher number of total episodes and infections, as a main effect and in interaction with attitudes to mother, hardiness, and alienation. Very few effects were obtained for traumas, suggesting that the effects of variables are mediated through a biological route rather than through behaviour, as would be the case if traumas were involved. No effect was shown by measures of stressful events alone. Results are discussed in the light of a reconsideration of the notion of stressful events. PMID- 8410749 TI - Personality characteristics and serum IgE level in patients with atopic dermatitis. AB - On the basis of clinical observations it was predicted that patients with atopic dermatitis (AD) differ from patients with chronic obstructive bronchitis (OB) in three personality variables; excitability (EX), inadequate stress coping (CO), and paranoidity (PA). Sixty-one patients with AD and 15 with OB were assessed using standard scales. Systematic comparisons between groups strongly confirmed these hypotheses. It was further assumed that AD patients with clinically relevant serum IgE levels (IgE > 100 IU/ml) score significantly higher on EX and CO than those with normal serum IgE levels (IgE < 100 IU/ml). In contrast, no significant differences between these subgroups were expected regarding paranoidity (which may be a consequence of the AD patients' cosmetic problems). These assumptions again were supported by the findings of the present study. The results underline the potential importance of assessing personality variables and serum IgE levels in the diagnosis of AD patients. PMID- 8410750 TI - Psychosocial factors in youth and at working age as predictors of blood pressure. A prospective study. AB - This prospective study focused on childhood and adult psychosocial determinants of blood pressure (BP) measured at working age, including work factors, individual characteristics and socioeconomic factors. Data had been collected over 20 years ago from a sample representative of Finnish children. For this follow-up, 150 subjects responded to a questionnaire and attended a personal examination at the mean age of 37 yr. Hierarchical regression analyses were made for women and men. A split half procedure was applied for the control of possible chance associations. Work conditions and perceived health were not associated with BP. Social conditions in youth were powerful predictors of high adult BP for men. Poor living conditions in youth explained 16% of their diastolic BP and 9% of their systolic BP. The result may be regarded as strong evidence for early determinants of hypertension. PMID- 8410751 TI - On physiological effects of positive and negative life changes--a longitudinal study. AB - A sample of working men and women were followed longitudinally on four occasions during a year with regard to serum triglycerides, plasma prolactin and systolic and diastolic blood pressure at rest. Participants were asked about life changes that they had gone through during the examination year. They were also asked to rate the importance and the direction (positive and negative, respectively) of the changes. On the basis of this information participants were divided into a no event group (N = 20), a positive event group (N = 11) and a negative event group (N = 11). Statistical analyses revealed that prolactin levels tended to increase in the negative event group whereas blood pressure as well as serum triglycerides increased in the negative event group and decreased in the positive event group. PMID- 8410752 TI - Appraisal and response to pain may be a function of its bodily location. AB - This study explored the hypothesis that appraisals of and responses to pain may in part be a function of where the pain is located. Results revealed that appraising pain as life-threatening, seeking help, disclosing pain to others and emotional distress in response to pain were contingent on the pain's bodily location. Significant ethnic differences in appraisal of pain, and significant gender differences in disclosure of and emotional response to pain also were found. PMID- 8410753 TI - Anger imagery and corrugator electromyography. AB - Electromyography (EMG) of corrugator muscle activity has been positively related to negative affective states and psychopathology. Research exploring the relationship between corrugator EMG and personality traits is lacking. The relationship between corrugator EMG and state-trait anger was examined using affective imagery in 52 undergraduate females divided into high and low trait anger groups. Results indicated that the high group reported greater difficulty with imagery, but reported more anger during the angry imagery as compared to the low anger group. Covarying imagery ability, no group differences in corrugator EMG were apparent. The greater experience of subjective anger by the high anger group was not associated with greater corrugator EMG or better imagery ability. PMID- 8410754 TI - Road map to a revolution: highlights from the 1993 APNA conference. AB - 1. However it may happen, change is certain to occur; mental health care will be reformed in the near future. 2. The Coalition of Psychiatric Nursing Organizations developed an 85-page position paper on essential services. One topic dealt with is "barriers to care," which include public attitudes, fragmented systems, poverty and race, private insurance, and public funding. 3. As a result of increasingly sophisticated and assertive lobbying efforts, nurses have been more involved in the political scene. Ira Magaziner, project director for the Health Care Reform Task Force, addressed 100 nurses recently. 4. For psychosocial nurses, "the future is now." There is a window of opportunity at this time, and nurses must agree on what is wanted and work together to take advantage of that window. PMID- 8410755 TI - The challenge of diversity. PMID- 8410756 TI - Mental health care reform: a historic town meeting of psychiatric nurses. AB - 1. Inspired by the 1992 presidential candidates' town meetings, the conveners of the American Psychiatric Nurses' Association Invitational Conference decided to stage a town meeting for psychiatric nurses. Forty psychiatric nursing leaders came together to share their professional expertise and experiences by giving "testimony" about the role psychiatric nurses play in mental health care reform. 2. The town meeting participants discussed many timely topics, such as practice, education, titling and credentialing issues, and the future of the psychiatric/mental health nursing profession. 3. The participants left the town meeting energized with a common sense of purpose--and a promise to commit themselves to creating the strategies necessary for reform of the mental health care system. PMID- 8410757 TI - Meet Mary Harper. Co-chair, health care reform, Mental Health Task Force. Interview by Kathryn K. Wheeler. PMID- 8410758 TI - New partnerships. Creating the future. PMID- 8410759 TI - All patients deserve fair care. PMID- 8410760 TI - Another look at codependency. PMID- 8410761 TI - A continuing journey: health care reform and mental health--to the Rose Garden, the White House, and more. PMID- 8410762 TI - The role of nursing in public policy reform. AB - Currently, individuals concerned about the cost and quality of health care, and the necessity of major system reform, are grappling with the concepts of managed competition, managed cooperation, sponsors, and health insurance purchasing cooperatives. Managed competition, broadly defined, is a purchasing strategy designed to obtain maximum value for consumers and employers as they acquire health care. Managed competition occurs at the level of integrated financial and delivery plans, not at the individual provider level. The goal of managed competition is to divide the providers in each community into competing economic units to motivate them to develop efficient delivery systems (Enthoven, 1993). Efficient health care delivery systems have not resulted from the fee-for-service and remote third-party reimbursement practices that currently dominate the health care system. However, more efficient delivery systems have emerged from managed care. Some of the best known models of managed care that have demonstrated cost effectiveness include Kaiser Permanente, Group Health Cooperative of Puget Sound, and Washington Group Health. The rapidly escalating consumption of US economic resources in the name of health care has led to the current pressure for health care reform. Although the public discussion about health care reform centers around the high cost of health care, we are, in truth, in the midst of both a crisis of medical care and an emergence of an era of health care for the nation's people. PMID- 8410763 TI - The concept of an 'Integrated Survival System' for protection against the responses associated with immersion in cold water. AB - In this paper the concept of an 'Integrated Survival System' (ISS) is introduced and discussed in relation to the helicopter passenger/crew member, although the principles are equally applicable to many other types of user and circumstance requiring specialized protective clothing. The fundamental principles behind this concept are first, that the wearer should be given protection against all of the hazardous responses associated with immersion in cold water and secondly, that the individual components which make up the ISS must be compatible and complementary; they may also be interdependent. PMID- 8410764 TI - A clinical trial to compare plaque removing efficiency of a prototype toothbrush with alternative toothbrushes. AB - Three alternative toothbrushes were investigated in a double blind Latin Square repeated measures design study for their relative efficiency at removing plaque 'in vivo'. A prototype filament suspension design toothbrush was tested, with the suspension mechanism both active and inactive, against a conventional toothbrush. Following an initial control plaque score, subjects were ordered by oral hygiene levels and randomly allocated to each of the three test groups. The trial was made up of three periods of 14 days consisting of four days of no brushing followed by 10 days of brushing with each of the three toothbrushes. All plaque scores were recorded using the Greene and Vermillion Index of Oral Hygiene and plaque scores were recorded for each patient at the start, four days later and at the end of each 14-day period. There were no significant differences in the quantity of plaque remaining after brushing with any of the brushes tested in this study. Ignoring the toothbrush used, the end brushing average plaque scores for period 1 were found to be higher than the control plaque scores. It is suggested that this resulted from the patients finding new toothbrushes less comfortable to use initially but following a period of accommodation and adjustment, end brushing plaque levels in subsequent periods returned to near the same level as the control plaque score. PMID- 8410765 TI - The management of alcohol abuse in the Royal Navy. AB - This paper offers a brief outline of the current Royal Naval policy on alcohol abuse. It starts by setting the Navy's problem in the context of a widespread social phenomenon. Attention is then drawn to the peculiar stressors of a military environment which may contribute to the development of problems in certain individuals. From this introduction, the practical implications of the policy are made explicit. This has two main strands; education and dealing with problem drinkers. The use of education to prevent the development of alcohol related difficulties at a general, whole force and specific divisional/management level is described. The identification and management of problem drinkers is also outlined. This is followed by a discussion of the policy which concludes with the listing of its strengths as perceived by its operators. PMID- 8410766 TI - Acclimatization to altitude: effects on arterial oxygen saturation and pulse rate during prolonged exercise at altitude. AB - Changes in the arterial haemoglobin-oxygen saturation (SaO2) of the blood, in the blood pressure and in the heart rate, were monitored in subjects performing static exercise at an altitude of 3,600 metres before and after an acclimatization period of 28 days during an expedition to the Bolivian Andes. It was found that acclimatised subjects could maintain their SaO2 during prolonged exercise better than non-acclimatised subjects. The pulse rate of acclimatised subjects was consistently lower at the same work level than that of non acclimatised subjects. Acclimatised subjects were able to reach higher levels of exercise than when they had first arrived at altitude. These data imply that acclimatization to altitude improves the delivery of oxygen to the tissues. The mechanism of this is not certain, but it may be concerned with a reduction in the degree of ventilation-perfusion inequality of the lung which occurs on exposure to high altitude. PMID- 8410767 TI - Malta and the British Navy: the medical connection during the nineteenth century. Part II. Some medical practitioners of note. PMID- 8410768 TI - Leptospirosis. PMID- 8410769 TI - The Royal Naval Medical Library Service. PMID- 8410770 TI - [New interpretation of the anatomy, physiology and pathology of the vertebral intersomatic joint: disk herniation does not exist]. PMID- 8410771 TI - [Secondary malignant tumors of the vestibulocochlear and facial nerves]. AB - INTRODUCTION: Neuromas and meningiomas represent by far the most frequent tumors of the cerebellopontine cistern (respectively over 80% and over 10%). Secondary malignant tumors are exceptional. METHOD: Presentation of four cases of malignant tumors of the cerebellopontine cistern, either revealing or complicating a primary tumor: a bronchial cancer, a cancer of the kidney, a medulloblastoma and a lymphoma. Patients were studied in T2- and T1-weighted high-field MRI sequences before and after gadolinium injection. RESULTS: The four tumors were enhanced by gadolinium and shown to invade the internal auditory canal, including one bilateral involvement. In most cases, the tumors were slightly heterogeneous and had irregular margins. CONCLUSION: These observations suggest that other diagnoses than acoustic neuroma should be taken into account. PMID- 8410772 TI - [MRI in infectious and spontaneous tumoral epidural hemorrhagic pathology]. AB - Several studies have served to underline the interest of MRI in the diagnosis and assessment of epidural abscess and malignant extradural spinal tumors. In our work, one of the quantitatively greatest published to this day, 64 patients presenting epidural pathology were evaluated by means of MRI. All the explorations were brought about through weighted multiplaned sequences T1, T2 and gadolinium injection. The pathological spectrum encountered is comprised of: metastatic extradural spinal tumors (58%), primary extradural spinal tumors (7%), epidural localizations of hematological diseases (9%), epidural abscess (25%), and one case of epidural hematoma. Assessment was brought to bear upon behavior in relation to type of sequence, spread of disease, medullary effects, type of enhancement following gadolinium injection. Degree of correlation with clinical data and surgical findings was also appraised. As concerns tumourous epidural pathology, positive diagnosis due to a lesion hinges upon the T1 and T2 sequences. Gadolinium's contribution is restricted to analysis of perivertebral and vertebral spreading; it also facilitates recognition of the spinal cord in circumferential epiduritis. So much said, gadolinium appears to be essential in frequently less compressive forms of infectious epiduritis; it plays a pronounced role in both follow-up of their evolution and recognition of acute epidural hematoma. PMID- 8410773 TI - [Echography in injuries of costal cartilages]. AB - Interest of high frequency ultrasonography in rib cartilage cannot be recognised by conventional X ray films: authors show the interest of high frequency ultra sonography when the trauma is known, finding a localised tumefaction, sometimes a rib cartilage fracture. When the traumatism in unknown, ultra sonography can correct the diagnosis. PMID- 8410774 TI - [Luxation of a hypoplastic posterior arch of the atlas. Report of a case]. AB - A rare case of sagittal dislocation of the hypoplastic right semi-posterior arch of the atlas, complicating a cervical trauma, is described. The authors specify its radiographic and CT appearance. This is a congenital abnormality discovered accidentally, for which no specific surgical treatment was implemented. PMID- 8410775 TI - [Clinical application of a digital method in mammographic image processing]. AB - The authors describe digital processing of mammographic images based on simulation of the procedure performed by radiologists to analyse radiographs. Image processing allows automatically enhancing contrasts and expressing them as a range of different light intensities and colors. Minimum contrast differences that are difficult to perceive with the bare eye are thus highly enhanced. This processing of mammographic images has proved useful not only to detect discrete lesions, but also to differentiate benign and malignant lesions. PMID- 8410776 TI - [Proposal of a method of automatic interpretation of images in x-ray computed tomography]. AB - A preliminary study is described relative to the possibility of using methods of automatic image interpretation in the field of biomedical images. These general methods are capable of recognizing characteristic shapes inside the image. In this paper, the structures are described and interpreted. These methods are capable of outlining shapes which are important from a clinical point of view- e.g., spine, vertebrae. They can also evaluate the shapes by plausible methods, to explain the meaning of the whole image. A feasibility study is described concerning the automatic recognition of vertebral contour in axial CT images. The results suggest that the employed techniques can be used in real diagnostic applications. PMID- 8410777 TI - [Diffuse peritoneal lymphoma in AIDS]. AB - We report an unusual observation of peritoneal localisation in a non-Hodgkin lymphoma, the only sign of digestive injury. This peritoneal injury may be more frequent with AIDS patients, as in our observation. Tomodensitometry was the best exam to visual this injury. PMID- 8410778 TI - [Intramural hematoma of the duodenum following acute pancreatitis. X-ray computed tomographic aspect of a case]. AB - Intramural hematomas of the duodenum in adults most often have a traumatic cause, but they may also be a complication of acute alcoholic pancreatitis. The diagnosis is based on the CT appearance. Treatment is usually medical. Surgery is necessary in case of recurrence or of a major hemorrhagic syndrome. PMID- 8410779 TI - [Emphysematous cystitis. X-ray computed tomographic aspect. Apropos of a case]. AB - Emphysematous cystitis is a rare urinary infection which is characterized by gas collection within the bladder wall, with or without gas in the lumen itself. The diagnostic is radiologic. We report a new case, discovered with C.T. and we discuss the interest of this technique. PMID- 8410780 TI - [The beginning of radiology in Montpellier]. PMID- 8410781 TI - [Role of x-ray computed tomography in the follow-up of Hodgkin's disease]. PMID- 8410782 TI - [The East African training center of radiologists in Nairobi (Kenya)]. PMID- 8410783 TI - Salvage of wounds following failed tissue transplantation. AB - During the past 20 years, 972 microvascular transplantations have been performed for 783 patients, with an overall failure rate of 6.2 percent. Fifty-four of the 60 failed transplantations were available for long-term follow-up and were retrospectively reviewed with respect to the original indications for transplantation, the number, and the type of salvage procedures performed following transplant failure. This study illustrates that the choice of salvage procedures performed following transplant failure depends on the original indications, the location, and the severity of the resultant wound. Failure following transplantation for coverage of contour defects or unstable wounds can often be managed by non-microsurgical methods. In contrast, when the indications for transplantation included the transfer of specialized tissues for thumb or digit reconstruction, the restoration of motor or sensory function, or the coverage of a limb-threatening wound, requirements for reconstruction could be satisfied only by a second successful tissue transplant. Eighteen of the 54 cases underwent an additional transplantation, with an 89 percent success rate. PMID- 8410784 TI - The use of cryopreserved venous allografts in microvascular surgery without immunosuppression: an experimental study. AB - Excellent patency rates are currently established in the use of autogenous veins as interposition vein grafts in microsurgical practice. Allografts may be a viable alternative source of vein grafts. Recent advances in cryobiology have enabled the controlled freezing of tissues with preservation of vital cellular elements. Although several reports have shown the successful use of cryopreserved large-vessel allografts, few have addressed cryopreservation of microvessels (1 to 2 mm in diameter). In this study, the authors have successfully cryopreserved femoral veins in a rat model and transplanted them as interposition vein grafts into arterial defects across major histocompatibility barriers (ACI to Lewis). Short-term patencies (21 days) were determined, and histologic and scanning electron microscopic analysis were performed. Patency of 100 percent was achieved in both fresh control veins and in cryopreserved veins at 21 days. PMID- 8410785 TI - The effect of a distal site of compression on neural regeneration. AB - The effect of a distal site of nerve compression on neural regeneration after a nerve repair was investigated in the rat model. After chronic compression had been established by placement of a Silastic band about a distal site of the posterior tibial nerve, the proximal posterior tibial nerve was divided, and then immediately repaired. Beginning 6 months after nerve repair, neural regeneration, assayed by walking track analysis, demonstrated significantly impaired function in the group of rats with a distal site of compression, compared with the repair group without a distal site of compression (p < .05). Followed for 3 additional months, neural function further significantly (p < .001) deteriorated in the group with distal compression. Morphometric analysis demonstrated that both the nerve repair alone and repair-plus-compression groups had 1) significantly decreased axon and nerve-fiber diameters (p < .001), and 2) significantly increased myelinated nerve-fiber density (p < .001), compared with normal; these findings are consistent with neural regeneration. However, the repair-plus compression group had significantly (p < .03) fewer regenerating fibers than did the repair-without-distal-compression group. This experimental study suggests that consideration be given to release of sites of known anatomic narrowing in close proximity to a nerve reconstruction. PMID- 8410786 TI - Anatomic variation in the blood supply of the radial forearm flap. AB - The traditional radial forearm flap derives its blood supply from perforators of the radial artery. This report describes an anatomic anomaly in which the flap was supplied solely by perforators of the ulnar artery. Knowledge of this previously undescribed variation might have relevance when a radial forearm flap is raised. PMID- 8410787 TI - Improved venous drainage of the radial artery forearm free flap: use of the profundus cubitalis vein. AB - The radial artery forearm free flap (RAFFF) is a versatile and popular flap used in various reconstructive procedures. This flap contains two venous drainage systems: a superficial (cephalic) route, and a deep (venae comitantes) route. Most surgeons use either the deep venous system alone or in combination with a superficial vessel, hence requiring two venous anastomoses. The authors describe a novel technique of providing venous drainage for the RAFFF, incorporating both deep and superficial systems, and using one venous anastomosis. From 10 consecutive operative and five cadaveric dissections, it was noted that paired venae comitantes, corresponding to the radial artery, coalesce into a single vein at the level of the brachial artery bifurcation. The profundus cubitalis vein (vena anastomotica) connects this coalesced (deep) vein to the cephalic (superficial) vein at the level of the cubital fossa. Both the deep and superficial venous systems are drained via one large anastomosis, situated proximal to the profundus cubitalis interconnection. Advantages of this technique include: 1) improved venous drainage of the RAFFF; 2) rendering the venous anastomosis technically easier, due to its larger size; and 3) more available versatility in designing the venous portion of the vascular pedicle. PMID- 8410788 TI - The pedicled arterialized venous flap. AB - The arterialized venous flap, in which arterial blood flowing through a vein returns to the venous system through the pedicle, was devised to solve the problems of increasing flap size and raising the overall success rate. This flap can obtain satisfactory blood inflow and pressure. The flap was clinically applied to reconstruct skin defects in three cases, with two complete successes and one partial superficial necrosis. This technique provides a flap to cover a relatively large defect, with no shunt formation, and a high success rate. PMID- 8410789 TI - Silicone cuffs for peripheral nerve repair: experimental findings. AB - The effectiveness of silicone cuffing in peripheral nerve repairs was assessed in a comparative study in rats. The femoral nerve was dissected out and severed bilaterally. The nerve ends were brought together with four epineural sutures on one side, while only one epineural suture was used on the other side, placing a silicone cuff around the junction site. Initially, nerve regeneration appeared to be improved by the cuff, with electrophysiologic parameters and histologic studies 6 weeks after the procedure yielding better results for the tubulated nerves. Subsequently, however, conductivity was less in the tubulated nerves, and results were clearly inferior to sutured nerves at 4 months after the operation. The deterioration was due to nerve compression caused by the cuff, which prevented axons from penetrating into the distal nerve stump, and resulted in axon fragmentation in the compressed nerve segment. The risk of nerve compression makes the use of silicone cuffs of doubtful value. PMID- 8410790 TI - The influence of magnitude and duration of crush load on functional recovery of the peripheral nerve. AB - This study investigated the effect of crush duration at different loads on motor functional recovery. One hundred and thirty-eight rats were divided into five groups: sham operation, resected sciatic nerve, and 100 g, 500 g, and 15,000 g of sciatic crush load (Groups 1 to 5). According to crush duration, Groups 3 and 5 were divided into 10-min, 2-hr and 6-hr subgroups. In Groups 3 to 5, a 5-mm segment of sciatic nerve was crushed, using a specially-designed crushing device. Motor functional recovery was assessed by calculating a sciatic functional index (SFI). There was no functional deficit in Group 1, and complete dysfunction in Group 2 throughout the experiment. All groups subjected to crush exhibited an initial complete deficit that gradually recovered to normal or near normal. Axonal damage and the speed of motor functional recovery were significantly related to crush duration in the subgroups of the 100 g group, but no marked differences existed between subgroups of the 15,000 g group. There was no obvious difference between the 6-hr subgroup of Group 3 and Groups 4 and 5. Results indicate that crush duration is an important factor in nerve damage and functional recovery at a low crushing level (100 g), and that the mechanical insult is a key factor at a higher crush level (15,000 g). The fact that all crushed nerves recovered, even after the application of a 15,000 g load for 6 hr, suggests the importance of maintaining continuity of the injured nerve in clinical situations. PMID- 8410791 TI - Toe-to-thumb transplantation in the rat. AB - A toe harvested from the hind foot of a rat was successfully transplanted to the rat thumb position, with vessel and nerve anastomoses. With this technique practiced, there will be more ease, confidence, and success when doing toe-to hand or small tissue transplantations or replantations in humans, even in infants. The procedure is described and discussed. PMID- 8410792 TI - Skeletal muscle fiber types and their vascular support. AB - Different neural and thyroidal influences on mammalian skeletal muscle result in considerable heterogeneity in muscle-fiber characteristics. Muscle fibers can, nonetheless, be grouped into three relatively homogeneous classes, based primarily on their contractile properties. There is a remarkable matching of metabolic support systems to contractile properties and, in turn, appropriate vascular supply for the metabolic systems of each of the three muscle fiber types. The contractile, metabolic, and vascular characteristics of each fiber type are consistent with known patterns of muscle fiber recruitment for the performance of muscular work. PMID- 8410793 TI - Interposition vein grafting in head and neck reconstructive microsurgery. PMID- 8410794 TI - Patterns of plasma LH, FSH, oestradiol and corticosteroids from birth to the first oestrous cycle in Meishan gilts. AB - Hormonal changes associated with pubertal development in a breed of pig exhibiting early puberty were determined. In Expt 1, blood from prepubertal Meishan gilts was collected at about 1 (n = 5), 10 (n = 5), 20 (n = 4), 30 (n = 5), 50 (n = 5), 70 (n = 10), 80 (n = 10) and 90 (n = 7) days of age. In Expt 2, females were sampled between 99 and 116 days of age before (n = 16) or during (n = 9) the first oestrous cycle. Serial blood samples were collected through a jugular catheter at 20 min intervals. In Expt 2, age at puberty was determined by daily examination for oestrus and weekly evaluation of plasma progesterone and found to occur at 111 days of age. Hormone profiles determined at least 19 days before the first oestrus were grouped to constitute a prepubertal stage. In Expt 1, mean concentration of LH was high at 1 day of age, significantly lower at 10 days and showed nonsignificant variations until 90 days. Frequency of LH pulses increased between 10 and 50 days (0.42 versus 0.85 pulse h-1, P < 0.05), decreased between 50 and 70 days (0.49 pulse h-1, P < 0.05) and remained low thereafter. Amplitude of LH pulses was high between 1 and 30 days and declined progressively until 80 days. Mean concentration of FSH was high between 1 and 20 days and decreased progressively until 80 days (7.2 and 2.3 ng ml-1, respectively, at 20 and 80 days, P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410795 TI - Influence of season on seminal characteristics, testis size and serum testosterone in the western spotted skunk (Spilogale gracilis). AB - The western spotted skunk is a seasonally breeding mammal: most copulations occur in late September and early October. This study was performed to characterize the seasonal changes in concentrations of testosterone and in ejaculate quality. Captive males (n = 22) were maintained on a natural photoperiod for 15 months. Semen samples were collected by electroejaculation; testis size was measured; and blood samples were collected. Of 110 electroejaculation attempts, 104 (95%) resulted in successful fluid collection and 101 (97%) samples contained spermatozoa. Significant increases (P < 0.05) in serum testosterone concentration, testis size and ejaculate volume were observed from August to November. Mean concentration of testosterone in serum ranged from 0.15 +/- 0.05 ng ml-1 in mid-January to 6.42 +/- 1.79 ng ml-1 in early October. Mean testis size ranged from 1.22 +/- 0.25 cm2 in February to 2.68 +/- 0.08 cm2 in October. Mean ejaculate volume ranged from 11 +/- 3 microliters in March to 129 +/- 22 microliters in October. Seasonal changes in the number of spermatozoa per ejaculate or motility of spermatozoa were not observed. Mean number of spermatozoa per ejaculate was 8.14 +/- 0.85 x 10(6) spermatozoa (n = 97); motility was 56 +/- 2.4% (n = 93); semen pH was 7.76 +/- 0.20 (n = 6); osmolarity was 394 +/- 13 mmol kg-1 (n = 10); and 70.3 +/- 1.5% of the spermatozoa were morphologically normal (n = 47).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410796 TI - Correlation of growth hormone secretion during pregnancy with circulating prolactin in rats. AB - Growth hormone (GH) concentrations were measured throughout pregnancy in rats. The effects of surgical stress, ovariectomy, and treatments with the antiprogesterone mifepristone (RU 486) or the antioestrogen tamoxifen on serum GH, progesterone and prolactin were studied. GH concentrations were low during the first 18 days of pregnancy, except on the morning of day 5, and increased progressively from day 19 reaching peak values on the mornings of days 21 and 22. Thereafter GH concentrations decreased progressively, reaching very low values at 24.00 h on day 22, in parallel with a rise in serum prolactin concentrations. Surgical stress, performed at 12.00 h on day 20 of pregnancy, diminished serum GH concentrations 10 min later, but these returned to values similar to those of the non-operated rats 1-24 h later. Surgical stress did not modify serum prolactin concentrations at any time. Ovariectomy performed on the morning of day 19 produced the expected fall in serum progesterone and a rise in prolactin which lasted until the night of day 20. Serum GH concentrations were significantly diminished with respect to controls on day 20 and the morning of day 21 and then increased. Treatment with mifepristone on day 19 produced a simultaneous rise in serum prolactin and a fall in serum progesterone and GH by 08.00 h on day 21. Treatment with tamoxifen on days 3 and 4, or given daily from day 17 onwards did not modify prolactin concentrations but diminished serum GH concentrations at 08.00 on day 5 and on days 19-22, with the exception of a peak on day 22 (08.00 h). Tamoxifen also decreased serum progesterone concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410797 TI - Sensitive sex determination assay applicable to bovine embryos derived from IVM and IVF. AB - A rapid, sensitive method for sex determination which uses nested, allele specific amplification of bovine zfx and zfy was developed. Accuracy of the sex determination assay was verified with lymphocyte-derived DNA from 24 cattle of known gender. Reliability and sensitivity of the assay in embryonic sex determination were examined by typing duplicate bovine embryo biopsies of 2-8 cells. Sex determination was possible in 38 of 40 biopsies, and agreement between biopsy assays occurred in 17 of 18 cases. Applicability of the sex determination assay was further verified by examining gender of 12 fetuses derived from sexed embryos. Embryonic and fetal gender determinations agreed in 11 of 12 cases with the twelfth yielding an ambiguous result from fetal ultrasound examination. PMID- 8410798 TI - Regulation of pulsatile secretion of prostaglandin F2 alpha from the ovine uterus by ovarian steroids. AB - Two experiments were conducted to determine how progesterone and oestradiol regulate pulsatile secretion of PGF2 alpha from the ovine uterus. In Expt 1, ovariectomized ewes received: (1) no treatment, (2) oestradiol, (3) progesterone, or (4) oestradiol and progesterone (n = 5 ewes per treatment group) to approximate the changes in steroids that occur during the oestrous cycle. Jugular venous blood samples were collected at 30 min intervals for 48 h beginning at 08:00 on day 14 of steroid replacement. Blood samples were collected from five intact ewes at a comparable time of the oestrous cycle for comparison. The number and magnitude of pulses in 13,14-dihydro-15-keto-PGF2 alpha (PGFM) in jugular venous blood samples were used to assess uterine secretion of PGF2 alpha. Experiment 2 was conducted as Expt 1, except that the progesterone replacement protocol was modified to duplicate more closely the temporal pattern of progesterone observed in intact ewes. Results were similar in both experiments. Intact ewes averaged 4.4 +/- 0.6 pulses per 48 h blood sampling period. The frequency of pulses was less in ovariectomized ewes (P < 0.05). The number of pulses was increased by progesterone treatment (P < 0.01); the number of pulses in ovariectomized ewes receiving progesterone replacement was similar to that observed in intact ewes. There was a tendency for oestradiol to have a positive effect on the number of pulses (P = 0.12). The magnitude of pulses in intact ewes averaged 419 +/- 38 pg ml-1 and was much less in ovariectomized ewes (P < 0.05) than in intact ewes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410799 TI - Distribution of lysophospholipids and metabolism of platelet-activating factor in human follicular and peritoneal fluids. AB - The distribution of lysophosphatidylcholine, lyso-platelet-activating factor and platelet-activating factor (PAF) was studied in human plasma and in follicular and peritoneal fluid. In plasma, peritoneal and follicular fluids, 51%, 87% and 89%, respectively, of the total lipids were found in the protein fraction (the density > 1.21 fraction). Two forms of lysophospholipids were identified in this fraction: one of high affinity and one of low affinity for albumin. The metabolism of PAF in human follicular fluid, peritoneal fluid and plasma was also investigated. PAF-acetylhydrolase activity was found in both peritoneal and follicular fluids which induced a time-dependent hydrolysis of [3H]PAF. The half life of PAF was estimated to be 7-12 min in plasma, 15-25 min in peritoneal fluid and approximately 2 h in follicular fluid. PAF-acetylhydrolase activity in embryo culture media supplemented with 10% serum was markedly inhibited by addition of commercial serum albumin. When 25 g albumin l-1 was added, 22% of [3H]PAF was hydrolysed h-1 compared with 72% in media without albumin. The concentrations of lysophosphatidylcholine measured in plasma, in follicular and peritoneal fluids were 252, 286 and 53 mumol l-1, respectively. The distribution of these lysophospholipids and the metabolism of PAF in the female genital tract fluids reported in the present study provide evidence for the involvement of these biologically active lipid mediators in a variety of reproductive processes including sperm-egg interactions and embryonic development. PMID- 8410800 TI - Functional and ultrastructural characteristics of two types of rat granulosa cell cultured in the presence of FSH or transforming growth factor alpha (TGF-alpha). AB - Granulosa cells were isolated from 15-day-old, 25-day-old or PMSG-primed rats and were separated by Percoll gradient (20 to 60%) into five fractions. The cells in fraction 2 were mostly small cells (6.96-9.57 microns) and fractions 3 and 4 had a relatively high population of large cells (10.96-13.05 microns) which were sorted to collect a pure population of large cells. Aliquots of small or large cells were cultured separately in serum-free defined DMEM/F-12 medium containing 50 ng FSH ml-1, or 10 ng transforming growth factor alpha (TGF-alpha) ml-1 for 3 days. In PMSG-primed rats, the large cells produced 3.2-fold more progesterone than did small cells (with FSH/TGF alpha: 255 +/- 35.0 versus 77.32 +/- 14.5 x 10(-6) ng, day 1). Large and small cells from 25-day-old rats produced similar amounts of progesterone (FSH/TGF alpha: 65.68 +/- 9.6 versus 78.25 +/- 12.3 x 10( 6) ng, day 1). In 15-day-old rats, large and small cells produced very low concentrations of progesterone (FSH/TGF alpha: 4.69 +/- 1.2 versus 2.66 +/- 1.0 x 10(-6) ng, day 1). Large cells from PMSG-primed rats had characteristics of steroidogenic cells, i.e. smooth endoplasmic reticulum and well-developed mitochondria with tubular cristae compared with small cells, whereas small and large cells from 25- and 15-day-old rats contained the regularly occurring organelles without the endoplasmic reticulum of the smooth variety and mitochondria with lamellar cristae. This study shows that the heterogeneity of granulosa cells is related to size, metabolic response to FSH, TGF-alpha or to both factors and morphological features, all of which may be associated with the transition from preantral to preovulatory stages of follicle differentiation. PMID- 8410801 TI - Reproductive performance in relation to uterine and embryonic traits during early gestation in Meishan, large white and crossbred sows. AB - Previous studies have shown that females of the Chinese Meishan breed and of their F1 cross with European Large White pigs are very prolific, producing about four more piglets per litter than control Large White females. The main cause of this prolificacy is enhanced prenatal survival for a given ovulation rate in Meishan and F1 females and this is controlled by genes of the mother, not those of the conceptus. The objectives of this study were to determine whether genotypic differences in embryo survival were apparent in the period immediately after attachment and to compare embryonic and uterine development at this time. Sows in their third parity (20 Large White, 14 Meishan, 25 Large White x Meishan F1 and 25 Meishan x Large White F1) were killed 20-22 days after mating and their reproductive tracts recovered for further study. There were significant differences between the purebred sows, and crossbred sows were approximately intermediate for the number of corpora lutea (20.7 +/- 0.9, 27.8 +/- 1.1, 22.4 +/ 0.8 and 23.3 +/- 0.8 for the four genotypes, respectively), the number of embryos (15.2 +/- 0.9, 23.4 +/- 1.1, 17.2 +/- 0.8 and 18.8 +/- 0.8, respectively) and the proportionate embryo survival (0.74 +/- 0.04, 0.84 +/- 0.04, 0.78 +/- 0.03 and 0.82 +/- 0.03, respectively). There was a negative association within genotype between embryo survival and the number of corpora lutea. Adjusting for the genotypic difference in the number of corpora lutea increased the genotypic differences in embryo survival.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410802 TI - Localization and characterization of EGF/TGF-alpha receptors on peri-implantation trophoblast in sheep. AB - Receptors for epidermal growth factor (EGF) have been identified on the ovine trophoblast as early as day 15 of gestation. A radioligand assay with 125I labelled EGF was used to detect high and low affinity binding sites on the trophoblastic and placental membranes. The binding of 125I-labelled EGF was inhibited by increasing concentrations of unlabelled EGF. Competition studies with other peptide hormones including transforming growth factor alpha (TGF alpha), insulin-like growth factor-I (IGF-I) and ovine placental lactogen confirmed the specificity of EGF/TGF-alpha for its receptor. Cross-linking experiments using disuccinimidyl suberate (DSS) revealed a radiolabelled band of relative molecular mass 170 kDa. Immunohistochemical localization of the receptors demonstrated their distribution on the epithelial layer cells. The presence of receptors for EGF/TGF-alpha suggests that these factors could be involved in the regulation of embryonic development and fetal growth. PMID- 8410803 TI - Relationship of secretion of GnRH in vitro to changes in pituitary concentrations of LH and FSH and serum concentrations of LH during lactation in sows. AB - The objectives of this study were (1) to determine whether release of GnRH in vitro was related to concentrations of LH and FSH in serum and pituitaries and to oestradiol in serum of sows at mid- or late lactation, and (2) to determine whether weaning at mid-lactation changes concentrations of these hormones from values expected at mid-lactation to values expected at late lactation. Multiparous crossbred sows were killed on day 14 (n = 5) or day 28 (n = 5) of lactation or on day 14 post partum after litters had been weaned on day 10 (n = 5). Blood samples were taken every 6 h for 4 days before sows were killed, and the preoptic suprachiasmatic area, medial basal hypothalamus, stalk median eminence, anterior pituitary and ovaries were collected at slaughter. Sows killed on day 14 after having their litters weaned on day 10 had more (P < 0.01) preovulatory follicles (> 6 mm in diameter) than lactating sows killed on day 14 or 28 (7.0 +/- 1.2 versus 0.2 +/- 0.1 and 1.5 +/- 0.8, respectively). Concentrations of LH, FSH and oestradiol in serum during 90 h before slaughter were greater (P < 0.05) in weaned sows and lactating sows killed on day 28 than in lactating sows killed on day 14 (LH: 0.72 +/- 0.3 and 0.68 +/- 0.3 versus 0.45 +/- 0.2 ng ml-1; FSH: 39.3 +/- 2.7 and 57.3 +/- 4.0 versus 28.8 +/- 1.6 ng ml-1; oestradiol: 10.9 +/- 1.6 and 5.6 +/- 0.7 versus 2.7 +/- 0.2 pg ml-1, respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410804 TI - Electron microscope observations on LH-induced oocyte maturation in Japanese quail (Coturnix coturnix japonica). AB - The aim of this study was to describe the temporal sequence of ultrastructural changes in the boundary between the preovulatory oocyte and its surrounding follicular wall during maturation induced by injection of LH. Female Japanese quail were injected with ovine LH (20 micrograms per bird) 10-12 h before the expected time of ovulation. The largest and second largest follicles were excised before or 1, 2, 4 or 6 h after injection. The oocyte and the surrounding follicular wall were processed for observations using light and electron microscopy. Before injection of LH, cytoplasmic projections of granulosa cells interdigitated with microvilli on the surface of the oocyte and formed spot desmosomes and gap junctions with the oolemma. Two hours after injection of LH, the germinal vesicles in the largest but not in the second largest follicles began to break down and membrane-bound vesicles increased in number and size in the surrounding germinal disc. The junctions between the oocyte surface and the granulosa cell projections started to dissociate and a perivitelline space began to develop, possibly as the result of an accumulation of fluids transported from the capillary sinus in the theca interna. The first maturation spindle was formed 4 h after injection of LH, whereas the first polar body and the second maturation spindle were formed 6 h after LH stimulation. These observations suggest that the dissociation of connections between the oocyte and granulosa cells 2 h after exposure to increased concentration of LH is the first process of oocyte maturation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410805 TI - Maturation-promoting factor (MPF) is responsible for the transformation of sperm nuclei to metaphase chromosomes in maturing bovine oocytes in vitro. AB - Bovine oocytes at the germinal vesicle stage were inseminated in Brackett and Oliphant's medium in the presence of BSA (10 mg ml-1), caffeine (5 mmol l-1) and heparin (10 micrograms ml-1). When oocytes were transferred into tissue culture medium (TCM)-199 containing 10% fetal calf serum (FCS) with or without 6 dimethylaminopurine (6-DMAP; 2 mmol l-1) 8 h after insemination and cultured for 15-40 h at 39 degrees C in 5% CO2 in air, 74-83% of oocytes were penetrated and polyspermy (67-80%) was common. At 40 h after culture in 6-DMAP-free medium, 65% and 63% of unpenetrated and penetrated oocytes, respectively, reached metaphase II or beyond. A few (6%) oocytes were activated and contained both male and female pronuclei. Sperm metaphase chromosomes were observed in 90% of the penetrated oocytes. Penetration by more than four spermatozoa greatly retarded the meiotic maturation of the oocyte. However, sperm chromosomes were never observed in oocytes cultured in 6-DMAP supplemented medium and oocyte maturation did not proceed beyond the stage of prometaphase I. These results demonstrate the possible participation of maturation-promoting factor in metaphase chromosome formation in spermatozoa. PMID- 8410806 TI - Maintenance of spermiogenesis by exogenous testosterone in rats treated with a GnRH antagonist: relationship with androgen-binding protein status. AB - The relationship of the testicular distribution and [3H]dihydrotestosterone binding capacity of androgen-binding protein (ABP) to the completion of spermiogenesis was examined in mature rats given daily injections of 25 or 250 micrograms kg-1 body weight of GnRH antagonist (GnRH-A; Ac-D(2), Nal1, 4Cl-D, Phe2, D-Trp3, D-Arg6, D-Ala10) for two weeks with or without subcutaneous implantation of 10 cm testosterone capsules. GnRH-A administration resulted in a dose-dependent suppression of serum FSH, which was partially prevented in the 250 micrograms GnRH-A kg-1 group by exogenous testosterone. The total testicular testosterone content and concentration of testosterone in seminiferous tubular fluid were equally suppressed in both groups of rats treated with GnRH-A and receiving the testosterone supplement. ABP concentrations in interstitial and seminiferous tubular fluid were normal in rats given the 25 micrograms GnRH-A kg 1 dose, and were increased (P < 0.05) by concomitant testosterone treatment. In contrast, ABP concentrations in interstitial and seminiferous tubular fluid were increased in rats given the 250 micrograms GnRH-A kg-1 dose. This effect was attenuated when exogenous testosterone was given. Although binding of [3H]dihydrotestosterone by ABP in seminiferous tubular fluid was not affected by GnRH-A treatment, with or without exogenous testosterone, it was reduced in interstitial fluid by GnRH-A in a dose-dependent manner, and partially restored by testosterone administration. While complete spermatogenesis was maintained in rats given 25 micrograms GnRH-A kg-1, the number of step 7 and step 19 spermatids were both reduced by 35%, and were not affected by testosterone implants.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410807 TI - Requirement for Ca2+ mobilization and increased prostaglandin production for maximal decidualization in rats and the involvement of angiotensin II. AB - Studies were performed to determine whether the inhibition of the decidual cell reaction induced by intrauterine infusion of the angiotensin converting enzyme inhibitor enalaprilat in rats is reversed by activation of Ca2+ influx. Influx of Ca2+ was shown to be stimulated by angiotensin II in endometrial cells in this study. Ovariectomized, adult female rats were sensitized for the decidual cell reaction with steroid treatments. For experiments in vivo, intrauterine infusions of enalaprilat alone, or in combination with the Ca2+ ionophore A23187, a synthetic diacylglycerol, and dioctanoyl-sn-glycerol (diC8), and PGE2 were initiated on the day of uterine sensitivity. Enalaprilat inhibited the increases in uterine PG concentrations and uterine weight that occur following infusion of the vehicle. Concurrent infusion of A23187 partially, but not completely, reversed the inhibition of uterine weight increase; diC8 did not affect the inhibition of enalaprilat. A23187 did not reverse the effects of enalaprilat on uterine PG concentrations. Concurrent infusion of A23187 and PGE2 fully reversed the inhibitory effect of enalaprilat on uterine weight. For experiments in vitro, endometrial stromal and epithelial cells were obtained from uteri on the day of sensitivity and maintained in suspension. Cytosolic free calcium concentration ([Ca2+]i) was monitored in cell suspensions by fluorescence spectrophotometry using the Ca(2+)-sensitive probe, indo-1. Angiotensin II induced a transient increase in [Ca2+]i of endometrial stromal cell suspensions, but not of epithelial cells; PGE2 did not increase [Ca2+]i in stromal or epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410808 TI - Synthesis and secretion of lipids by bovine oviduct mucosal explants. AB - The objectives of this study were to determine the types of lipid synthesized and secreted by the bovine oviduct, and to determine whether lipid synthesis and secretion varied with stage of the ovarian cycle and oviductal region. Oviduct explant cultures were prepared from cows killed during either the follicular or luteal stage of the oestrous cycle. Both stage of ovarian cycle and oviductal region affected lipid synthesis by oviductal explants in vitro. More lipid was synthesized by explants from follicular than from luteal-stage cows. Ampullar explants synthesized the greatest quantity of total lipid, followed by the preampulla and isthmus. Separation of extracted lipids from cultured tissue by high performance thin-layer chromatography (HPTLC) resolved phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, cardiolipin, free cholesterol, free fatty acid, triglyceride and esterified cholesterol, all of which were synthesized during culture. The ampulla synthesized significantly more phosphatidylethanolamine, phosphatidylserine and phosphatidylinositol than did the other regions. Culture supernatants from ampullary explants contained the most newly synthesized cholesterol when compared with other regions. The histochemical location of neutral lipid droplets in the epithelium of cultured explants paralleled the localization of radioactivity in autoradiographs of explant extracts. The results suggest that the oviduct synthesizes a variety of lipids, and that some of these are released into culture supernatants. PMID- 8410809 TI - External ionic conditions, internal pH and motility of ram and boar spermatozoa. AB - Internal pH and motility of testicular, epididymal and ejaculated ram and boar spermatozoa were studied as a function of external ionic composition. Internal pH was estimated by the amine distribution method and motility was characterized by percentage of cells that were motile and flagellar beat frequency. Upon dilution in media at different external pH values, internal pH of boar and ram spermatozoa changed rapidly towards the external pH. High external concentrations of Na+ or K+ had no effect on the rate of equilibration and only a slight effect on the final internal pH value, ruling out a role of Na(+)-H+ or K(+)-H+ exchange mechanisms in this process. In both species, a linear relationship was observed between internal and external pH but equilibration was incomplete suggesting that there is a complex regulatory mechanism. This result was unaffected by epididymal maturation and ejaculation. Ram and boar testicular spermatozoa showed no increase in movement after dilution, suggesting that simple changes in internal pH are not a sufficient trigger for motility. At high external pH, internal pH increased and motility of epididymal boar spermatozoa was initiated. Motility of ejaculated boar spermatozoa, and epididymal and ejaculated ram spermatozoa was less dependent upon external pH and affected only very slightly by the internal pH changes. K+ or Na+ had almost no effect on motility just after dilution. After 1 h of incubation, movement decreased. Maintenance of motility in sodium or potassium showed a sharp external pH optimum. Media without Na+ and K+ allowed a better conservation of motility at external pH > 8 for ram epididymal and ejaculated spermatozoa and at external pH > 6 for boar ejaculated spermatozoa. PMID- 8410810 TI - Failure of luteolysis and extension of the interoestrous interval in sheep treated with the progesterone antagonist mifepristone (RU 486). AB - The progesterone antagonist mifepristone (RU 486) was injected i.m. into ewes during the early luteal phase of the oestrous cycle to test the hypothesis that duration of uterine exposure to progesterone from the corpus luteum initiates luteolysis through the proper timing of endometrial oxytocin receptor expression and pulsatile secretion of PGF2 alpha coincident with release of luteal oxytocin. In Expt 1, duration of cycle, the PGF2 alpha metabolite 15-keto-13,14-dihydro PGF2 alpha (PGFM) and oxytocin concentrations were measured in ewes treated on days 5, 6, 7 and 8 of the oestrous cycle with either 2.5 or 5.0 mg RU 486 kg-1 day-1 (n = 4 per group); control ewes (n = 6) were injected i.m. with 80% ethanol (diluent). In Expt 2, the presence of functional uterine oxytocin receptors was determined indirectly on day 12 of the cycle by measuring the plasma PGFM response to oxytocin challenge (20 iu, i.v.) in diluent-treated ewes (n = 3) and in ewes treated with 2.5 mg RU 486 kg-1 day-1 on days 6, 7 and 8 of the oestrous cycle. Duration of the oestrous cycle of control ewes (16 +/- 1 days) was extended beyond day 24 (day 0 = oestrus) in 10 of 11 ewes treated with RU 486 as determined by daily exposure of ewes to a ram and by measurement of progesterone concentrations in plasma in the two experiments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410811 TI - Vitrification of preimplantation stages of mouse embryos. AB - Three vitrification solutions (VS) namely VS1 (5.5 mol ethylene glycol l-1 and 2.5 mol glycerol l-1), VS11 (6.0 mol ethylene glycol l-1 and 1.8 mol glycerol l 1) and VS14 (5.5 mol ethylene glycol l-1 and 1 mol sucrose l-1) were tested for cryopreservation by vitrification of all developmental stages of mouse preimplantation embryos. In these experiments all preparative work was at room temperature (25 degrees C). VS1 was toxic to embryos at and earlier than the eight-cell stage, whereas VS11 was toxic to the four-cell and earlier stages. VS14 was the least toxic VS. All three VS resulted in good viability of vitrified Swiss Outbred day-4 embryos (morulae, early blastocysts and blastocysts) in vitro and vitrification with VS14 resulted in no loss of viability in all preimplantation stage Swiss Outbred embryos except one-cell embryos. One-cell F1 embryos were vitrified successfully with VS14 and VS1. The minimal equilibration time essential for successful vitrification of embryos suggests that concentration of the intracellular solutes by dehydration has a major role in establishing conditions conducive to intracellular vitrification. Studies in vitro suggested that sucrose dilution was not necessary in the removal of cryoprotectant from vitrified eight-cell and day-4 mouse embryos but, in contrast, development of vitrified day-4 embryos in vivo was better when the VS was diluted with sucrose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410812 TI - Biopsy and sex determination by PCR of IVF bovine embryos. AB - Sex of early bovine embryos was determined by polymerase chain reaction (PCR) using a single blastomere removed at the 16-32 cell stage. Embryos were produced in vitro and biopsied on the fifth day after in vitro fertilization. Biopsied embryos were cultured on a cumulus cell monolayer until embryo transfer. For the PCR, one pair of bovine-specific and one pair of Y-chromosome-specific primers were used. Definite signals following PCR amplification were obtained in 95.4% of cases indicating that one blastomere from a preimplantation bovine embryo is sufficient for sex determination by PCR. Nineteen biopsied embryos of predetermined sex were transferred into synchronized recipient females to examine their developmental potential in vivo. Ten of the recipients (52.6%) were found to be pregnant by ultrasonography 25 days after transfer. This result did not differ significantly from that achieved with the use of the control non manipulated IVF embryos (54.1%; P > 0.1). PMID- 8410813 TI - Circannual inter-relationships among reproductive hormones, gross morphometry, behaviour, ejaculate characteristics and testicular histology in Eld's deer stags (Cervus eldi thamin). AB - Blood samples, morphometric measurements and behavioural data were collected weekly for 52 weeks from six adult Eld's deer stags exposed to natural fluctuations in photoperiod (38 degrees N latitude). Mean (+/- SEM) serum LH concentrations reached peak values in the autumn (October), three months before FSH and testosterone concentrations reached peak values in early winter (January). Prolactin concentrations were inversely related (r = -0.733, P < 0.001) to LH and directly related to daylength, and maximal concentrations were observed during mid-summer (July) and minimal concentrations during early winter (January). The temporal pattern of circulating inhibin was positively correlated with FSH (r = 0.88, P < 0.001), but lagged behind the seasonal FSH increase by 1 3 weeks. Antler length, body weight and chest girth were maximal during pre-rut (December-January). Maximal scrotal circumference and combined testes volume were observed in mid-winter (February), whereas peak neck girth and behavioural aggression occurred 1-3 months later (March-May). On the basis of quarterly electroejaculation results, motile spermatozoa were produced in all seasons. However, the greatest number of motile spermatozoa per ejaculate was observed during the winter and spring, whereas the highest incidence of sperm pleiomorphisms (> 80%) was detected in the autumn. Histological assessments of the regressed testis (July) revealed fewer germ cells undergoing spermatogenesis and an increased incidence of degenerating and abnormal cell types. In summary, Eld's deer exhibit a circannual hypothalamic-pituitary-gonadal cycle with onset of pituitary activation occurring during the autumn and winter, whereas gonadal activity peaks during the winter and spring as daylengths are increasing. Marked circannual variations in circulating prolactin suggest that Eld's deer may use photoperiodic cues to modulate seasonal fertility; however, the existence of an endogenous seasonal rhythm operating independently of photoperiod cannot be excluded. PMID- 8410814 TI - Seasonal patterns of basal and GnRH-induced LH, FSH and testosterone secretion in Eld's deer stags (Cervus eldi thamin). AB - Plasma LH, FSH and testosterone were measured in blood samples collected via remote catheterization from six adult Eld's deer stags every 10 min for 8 h before and 2 h after GnRH (1 microgram kg-1, i.v.) administration. Blood samples were collected within 2 weeks of the summer solstice (21 June), autumn equinox (22 September), winter solstice (21 December) and spring equinox (20 March). Marked seasonal variations in basal LH, FSH and testosterone concentrations were observed. From autumn, well-defined LH pulses were temporally associated with small, but detectable pulses in testosterone. During the winter transition into the breeding season, episodic LH pulses were also temporally associated with corresponding testosterone surges that lasted 2-3 h. High amplitude, low frequency testosterone surges were also observed during the spring, but often in the absence of detectable LH pulses. Basal LH and testosterone concentrations decreased during the summer and, although LH pulses were detected, associated testosterone pulses were absent. Only 37% of LH pulses occurred coincidentally with FSH pulses, and FSH pulses were generally less prominent. The increases in LH and FSH above basal concentrations after GnRH treatment were significant (P < 0.05) for all seasons. Increases in testosterone after GnRH treatment were greatest during the winter and spring, but testosterone also increased to a lesser extent during the autumn (P < 0.05). In contrast, testosterone concentrations were not different before and after GnRH treatment during summer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410815 TI - Natural killer cytotoxicity and antibody-dependent cytotoxicity of cells of rat metrial glands. AB - Cell suspensions of rat metrial gland tissue, containing healthy granulated metrial gland (GMG) cells, were assessed for their ability to lyse natural killer (NK) cell target Yac-1 myeloma cells in a [51Cr]-release cytotoxicity assay. Cytotoxicity indices were found to be low even after in vivo stimulation of NK cytotoxic activity by polyinosilic-cytidilic acid and the addition of interferon to test cultures. It is concluded that cell suspensions of rat metrial gland, and rat GMG cells in particular, do not exhibit NK cytotoxicity. However, when a heat inactivated rabbit anti-Yac-1 serum was added to assay cultures, cytotoxicity indices above background levels were obtained, indicating that rat metrial glands contain cells capable of antibody-dependent cytotoxicity. PMID- 8410816 TI - Endocrine responses and conception rates in fallow deer (Dama dama) following oestrous synchronization and cervical insemination with fresh or frozen-thawed spermatozoa. AB - In Expt 1, 59 mature fallow deer does were allocated to six treatments (n = 9-10 per treatment). Does assigned to treatments 1, 2 and 3 each received an i.m. injection of 500 micrograms cloprostenol on day 13 of a luteal cycle. Does in treatments 2 and 3 received 50 or 100 iu pregnant mares' serum gonadotrophin (PMSG), respectively, at the time of prostaglandin administration. Does assigned to treatments 4, 5 and 6 each received single intravaginal controlled internal drug release (CIDR) devices for 14 days. Does in treatments 5 and 6 received 50 or 100 iu PMSG, respectively, at the time of CIDR device withdrawal. Incidence of oestrus was higher following treatment with CIDR devices than with prostaglandin (29 of 30 versus 12 of 29, P < 0.001). PMSG induced earlier onset of oestrus (34.6 +/- 0.9 h versus 44.7 +/- 2.4 h, P < 0.01) and reduced the range in the time to onset of oestrus (from 22 to 8 h for prostaglandin-treated does and from 36 to 14 h for progesterone-treated does). The number of LH surges was higher following treatment with CIDR devices than with prostaglandin (10 of 12 versus 3 of 12, P < 0.01). The overall mean peak LH concentration and time to LH peak were 30.2 +/- 3.4 ng ml-1 and 45.2 +/- 2.2 h after prostaglandin administration or CIDR device withdrawal. The overall median time of ovulation was 26 h after the onset of oestrus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410817 TI - Association of immunolocalization of matrix metalloproteinase 1 with ovulation in hCG-treated rabbit ovary. AB - Matrix metalloproteinase 1, MMP-1, which was previously called interstitial collagenase, is necessary for extracellular matrix reconstruction. The immunolocalization of the latent form of MMP-1 (proMMP-1) was examined in the ovulatory process in hCG-treated (100 iu per animal) rabbit ovaries. Immunoreactive products of proMMP-1 were identified by the avidin-biotin peroxidase complex formation using an anti-rabbit proMMP-1 polyclonal antibody. ProMMP-1 was distributed in the cytoplasm of theca interna cells, theca externa cells, interstitial glands and germinal epithelium throughout ovulation. However, at 9 or 10 h after hCG treatment, this enzyme was identified in several capillary lumina around the apex of preovulatory follicles. In addition, the staining density of immunoreactive products apparently increased in granulosa cells and theca interna cells around the orifice of the ruptured follicle 10 h after the stimulation. These results indicate that the spatiotemporal appearance of proMMP 1 in ovulation may be closely associated with the initiation of rupture of the follicle. PMID- 8410818 TI - Inhibition of gonadotrophin release in mares during the luteal phase of the oestrous cycle by endogenous opioids. AB - Effects of the opioid antagonist naloxone on concentrations of LH and FSH in plasma were measured in mares during different stages of the oestrous cycle. During the follicular phase of the cycle, naloxone (300 mg i.v.) had no discernible effects on basal concentrations of LH and FSH. A significant increase in plasma LH (P < 0.01) and FSH (P < 0.05) concentrations was observed after naloxone in mares during the luteal phase. This response was not different between suckled and non-suckled mares. The gonadotrophin-releasing hormone analogue buserelin (0.02 mg i.v.) caused a significant (P < 0.05) LH and FSH release irrespective of the stage of the oestrous cycle and a previous naloxone treatment. The results of this study indicate that endogenous opioid peptides are involved in the inhibition of LH and FSH release during the luteal phase of the oestrous cycle in mares and may partially mediate the suppressive influence of progesterone on gonadotrophin secretion. The opioid-mediated suppression of LH and FSH release does not seem to be affected by suckling. PMID- 8410819 TI - Effect of photoperiod on the breeding season of the marsupial possum Trichosurus vulpecula. AB - The production of young by three groups of eight possums, housed in a natural, a short-day (10 h light:14 h dark) and a long-day (14 h light:10 h dark) photoperiod was monitored for 24 months to determine the role of photoperiod on the occurrence and duration of the breeding season. Possums were housed in each of the three light regimens on 22 November. The possum has a gestation duration of 17.5 days. Removal of the newborn young leads to ovulation approximately 9 days later and birth 26 days later. Repeated removal of pouch young was used to determine the duration of the breeding season. Possums in a natural photoperiod showed two breeding seasons for the 2 year observation, from March to October of each year. Over the same 2 year period, there were three breeding periods in possums in short-day and long-day photoperiod. A comparison of the timing of the first breeding season showed that short-day photoperiod brought the breeding season forward by 2 months and long-day photoperiod delayed it by 2.5 months. The total number of months in which births occurred was the same for natural and short-day photoperiod (16 out of 24 months) and less for long-day photoperiod (11 months). Fertility in the three groups was also examined. Within each group, there was no change in fertility with consecutive seasons.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410820 TI - Oxytocin receptor binding activity in cultured ovine endometrium. AB - Oxytocin receptor binding activity in explants of caruncular and intercaruncular endometrium collected from luteal phase ewes increased during culture. An initial rise in binding activity occurred during the first 24 h of culture in both tissues; binding activity in intercaruncular endometrium continued to increase until day 6, remained unchanged on day 8 and had decreased by day 10 of culture. The maximum concentration of receptors in caruncular endometrium was significantly lower than that in intercaruncular endometrium (P < 0.001) and did not change significantly between days 4 and 10 of culture. Apparent dissociation constants and maximal binding of oxytocin receptor in cultured caruncular and intercaruncular endometrium were 3.09 and 2.72 nmol l-1 and 249 and 459 fmol [3H]oxytocin bound mg-1 protein, respectively. Concentrations of oxytocin receptor remained constant in myometrium during 96 h of culture. The rise in endometrial oxytocin receptor concentration did not result from exposure to fetal calf serum, phenol red or insulin in the culture medium. Substituting fetal calf serum with sheep serum or BSA did not block the rise in receptor binding activity. Actinomycin D and cycloheximide inhibited the rise in receptor concentration in both tissues. Co-culture of lung or kidney with endometrium had no effect on binding activity, whereas co-culture with luteal tissue effectively reduced the rise in oxytocin receptor concentration. To establish whether synthesis of functional oxytocin receptors occurred during culture, the effect of oxytocin on prostaglandin F2 alpha (PGF2 alpha) production was assessed in fresh tissue and after 48 h in culture.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410821 TI - Effects of the protein phosphorylation inhibitor genistein on maturation of pig oocytes in vitro. AB - In vitro maturation of cumulus enclosed and denuded pig oocytes was reversibly inhibited by the protein kinase inhibitor genistein. The half-maximal effect on maturation was observed at 40 micrograms ml-1. Genistein inhibited total protein phosphorylation and synthesis with the same dose-response relationship (ED50: 40 micrograms ml-1). Protein phosphorylation and synthesis patterns were changed by effective concentrations of genistein. Pig oocytes were sensitive to genistein during the first 12 h of in vitro maturation. This genistein sensitive period corresponds closely with the period of sensitivity to the protein synthesis inhibitor cycloheximide. Whereas the inhibition of protein synthesis affects only nuclear membrane breakdown and not chromatin condensation, genistein inhibits both events. The results of these experiments suggest that protein phosphorylation and synthesis play major roles during pig oocyte maturation in vitro. It is concluded that genistein inhibited protein phosphorylation is a regulator of chromatin condensation, whereas both new protein synthesis and phosphorylation appear to be required for nuclear membrane disassembly. Caution about this second conclusion is, however, necessary because of the dual action of genistein on both protein phosphorylation and indirectly on protein synthesis. PMID- 8410822 TI - Nuclear transplantation of embryonic stem cells in mice. AB - The developmental ability of enucleated mouse eggs that had received embryonic stem cells was examined. In a preliminary study, none of the reconstituted eggs formed a nucleus using inactivated Sendai virus (HVJ) after activation with ethanol. DC pulses were applied at 100 or 140 V mm-1 in addition to this treatment. After electrofusion, 29-40% of reconstituted eggs formed nuclei and 41 60%, 25-44%, 12-24% and 12-18% developed to two-cell and four-cell and morulae and blastocysts, respectively. The ability of reconstituted eggs to form blastocysts did not depend on the age of recipient eggs, except in cases of very young and very old eggs. Although implantation sites were observed, no live fetuses were obtained after the transfer of reconstituted eggs to the recipients. PMID- 8410823 TI - Effect of platelet-activating factor (PAF) on human spermatozoa-oocyte interactions. AB - The effect of exogenous platelet-activating factor (PAF) on human spermatozoa was examined by monitoring acrosomal status and hamster oocyte penetration. Induction of the acrosome response by PAF was dose, and time, dependent and required capacitated spermatozoa. Addition of 10(-14) mol PAF l-1 enhanced the acrosome reaction eightfold compared with controls treated with human serum albumin (HSA). Similar concentrations of lyso-PAF failed to induce acrosomal loss and preincubation of spermatozoa with CV3988, an inhibitor of PAF, prevented PAF induction of the acrosome reaction. PAF significantly increased the number of zona-free hamster oocytes penetrated compared with controls (9.8 +/- 0.5 of 25 oocytes were penetrated by control spermatozoa compared with 23.3 +/- 0.8 out of 25 oocytes penetrated after incubation of spermatozoa with 10(-14) mol PAF l-1; 93% of all oocytes were penetrated by at least one spermatozoon following incubation with PAF), and also increased the number of decondensed spermatozoa found per egg during the sperm penetration assay (from 1.7 +/- 0.3 spermatozoa/egg with control spermatozoa to 3.3 +/- 0.5 spermatozoa/egg with PAF treated spermatozoa). PAF-induced increases in acrosome reaction and sperm penetration assay values were similar to effects obtained with human follicular fluid and were calcium dependent. Induction of the acrosome reaction by physiological concentrations of PAF appeared to be morphologically similar to the response induced by follicular fluid as assessed by electron microscopy. PMID- 8410824 TI - Binucleate blastomeres in preimplantation human embryos in vitro: failure of cytokinesis during early cleavage. AB - The nuclei of disaggregated blastomeres from two hundred preimplantation human embryos were examined between days 2 and 4 after insemination in vitro by vital labelling with a polynucleotide-specific fluorochrome. Although the majority of blastomeres had a single nucleus, binucleate blastomeres containing two nuclei of equal size were common and other blastomeres had fragmented nuclei or were anucleate. Seventeen per cent of normally fertilized embryos at two- to four-cell stage had at least one binucleate blastomere, and this increased to 65% at the nine- to 16-cell stage when individual embryos had between one and six binucleate blastomeres. The proportion of binucleate blastomeres in normally fertilized embryos increased from 5 to 10% over this period, whereas in abnormally fertilized, polyspermic or parthenogenetic, embryos the proportion was significantly higher during early cleavage stages but decreased at the nine- to 16-cell stage when the majority of these embryos arrest (25 and 6%, respectively). The incidence of anucleate blastomeres in normally fertilized embryos was also high, especially in those of poor morphology. In contrast, blastomeres with fragmented nuclei were relatively uncommon and the incidence was variable among classes and stages of development. Estimates of the volume of binucleate blastomeres based on measurement of their diameters and comparison with mononucleate blastomeres at various cleavage stages indicated that these blastomeres arise from a failure of cytokinesis between the second and fourth cleavage divisions. On this basis, assignment of binucleate blastomeres to particular cleavage stages in normally fertilized day 4 embryos suggests that at least some of these blastomeres arising during early cleavage persist without further cell division for up to 48 h. At the cellular level, therefore, blastomeres with either binucleate or abnormal nuclei contribute to cleavage stage arrest in vitro. PMID- 8410825 TI - Effects of ovariectomy and genotype on bioactive FSH in plasma and pituitary of Booroola ewes. AB - Blood samples were collected for 13 days before and 20 days after ovariectomy from carrier (BB) and non-carrier (+ +) ewes of the Booroola FecB gene (n = 12 per genotype), at known stages of the oestrous cycle, after which the pituitary glands from these ewes were recovered. Pituitary glands were also collected from cyclic ewes (about day 12; n = 5 per genotype) to compare the effects of ovariectomy on pituitary gonadotrophins. Plasma samples and pituitary extracts were assayed for bioactive (B) FSH, immunoreactive (I) FSH and I-LH. Overall, BB ewes had significantly (P < 0.05) higher plasma I-FSH concentrations than did + + ewes before ovariectomy; the mean value was higher on 16 of the 17 days of the oestrous cycle (P < 0.01). For B-FSH, there were no overall genotypic differences, although the mean for the BB ewes was significantly higher on 13 of the 17 days of the oestrous cycle (P < 0.05), and significantly (P < 0.05) higher between days 13 and 16. No genotypic differences were noted for the plasma bioactive:immunoreactive (B:I) ratio for FSH before ovariectomy. After ovariectomy, there were significant (P < 0.001) increases in plasma for B-FSH, I FSH and I-LH and a significant (P < 0.05) decrease in the B:I ratio for FSH, irrespective of genotype.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410826 TI - Leucocyte distribution in the uterus during the preimplantation period of pregnancy and phagocyte recruitment to sites of blastocyst attachment in mice. AB - Antibodies to antigen F4/80 were used to identify monocyte-lineage cells in the preimplantation uterus and within and between early implantation sites, in intact pregnancy and using a model of delayed implantation, in mice. Neutrophils were identified by morphology and lymphocytes by immunocytochemistry and morphology. Macrophages and neutrophils were found throughout the endometrium on day 2 of pregnancy but were concentrated under the luminal epithelium. Macrophages were evenly distributed throughout the endometrium on days 3-4. Neutrophils were rarely seen on days 3-4, but lymphocytes were found throughout the endometrium, often in groups associated with glands or the luminal epithelium. Within attachment sites, a group of neutrophils was always seen associated with the luminal epithelium within implantation chambers on day 5 or, in ovariectomized, progestagen-maintained animals, at 20 h after a nidatory dose of 20 ng oestradiol. Macrophages were scarce in the subepithelial region of attachment sites and were absent from decidual tissue. 5C6, a monoclonal antibody that binds to a functional epitope of the type 3 complement receptor (CR3), a leucocyte adhesion integrin, was given before nidatory oestradiol. Treatment with 5C6 failed to modify the neutrophil recruitment to attachment sites and had no effect on the implantation rate in ovariectomized, progestagen-maintained animals given nidatory oestradiol. PMID- 8410827 TI - Quantitative and cytochemical studies of mast cell proteases in rat ovaries and uteri in various reproductive states. AB - A role for mast cell proteases (RMCP I and II) in the cyclical remodelling of ovarian and uterine tissues of rats was investigated in the oestrous and pregnancy cycles using immunocytochemistry and enzyme-linked immunosorbent assays. The concentrations of RMCP I exceeded that of RMCP II by 100-fold in both tissues, but were always much higher in uteri than ovaries. Most of the protease activity in the uterus was located in the myometrium, whereas it was more focally distributed in the hilus and medulla of the ovary. Protease activity was confined to mast cells identified by metachromatic staining and no single cell contained both proteases. The concentrations of RMCP I and II in the two organs did not fluctuate throughout the 4-day oestrous cycle. Neither were RMCP I concentrations in the uterus altered by administration of diethylstilboestrol to ovariectomized animals, although total amounts per uterus were substantially greater than in the controls. Concentrations of RMCP I were substantially reduced in the uterus after day 6 of pregnancy and rose during the puerperium. The reduction was greater in pregnant than in pseudopregnant horns and tended to be lower in the vicinity of conceptuses than between them. The physiological significance of the lower mast cell protease concentrations is unclear, although their absence may contribute to the decreased protein catabolism during pregnancy. PMID- 8410828 TI - Production of matrix metalloproteinase 1 (interstitial collagenase) and matrix metalloproteinase 2 (gelatinase A: 72 kDa gelatinase) by ovine endometrial cells in vitro: different regulation and preferential expression by stromal fibroblasts. AB - Ovine endometrial cells (epithelial plus stromal), prepared from ovariectomized ewes treated with oestrogen and progesterone to mimic the luteal phase of the oestrous cycle were maintained in serum-free medium for 48 h in the presence or absence of phorbol myristate acetate (PMA, 100 nmol l-1), a known stimulus for production of matrix metalloproteinases (MMP) in other cells. Matrix metalloproteinase-1 (MMP-1, interstitial collagenase) and matrix metalloproteinase-2 (MMP-2, gelatinase A) activities were expressed by the cells in the absence of PMA; most were in the latent form and required activation by (4 aminophenyl) mercuric acetate (APMA). Exposure to PMA over 48 h resulted in a significant increase in MMP-1 activity but only a modest and nonsignificant increase in MMP-2 activity. Gelatin zymography demonstrated that proMMP-2 (72 kDa) was produced by both PMA-treated and untreated cells and an active form of 67 kDa was also present. Immunolocalization of MMP-1 and MMP-2 was seen within the cells following treatment with monensin. Highly purified epithelial and stromal cells were similarly cultured and analysis of the conditioned medium showed that MMP-1 and MMP-2 were produced predominantly by stromal rather than epithelial cells. Thus, both MMP-1, which degrades interstitial collagens, and MMP-2, an important enzyme for degradation of type IV and V collagens, are synthesized and released by ovine endometrial stromal cells in culture, but MMP-1 is produced primarily upon stimulation, whereas MMP-2 production is constitutive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410829 TI - GnRH-dependent and -independent components of FSH secretion after acute treatment of anoestrous ewes with ovine follicular fluid and a GnRH antagonist. AB - Gonadotrophin and inhibin concentrations were measured in anoestrous ewes after acute treatment with either saline, ovine follicular fluid (oFF), GnRH antagonist, or oFF and GnRH antagonist in combination. The increase in mean LH concentrations observed in ewes treated with oFF alone, was not seen in either of the groups treated with GnRH antagonist, in which LH pulsatility was completely inhibited. This result suggests that the LH rebound that follows follicular fluid treatment is GnRH dependent. Blockade of GnRH had no effect on the suppression of FSH seen after follicular fluid injection, indicating that this component of FSH secretion is independent of short-term GnRH input. After this initial suppression, a rebound release of FSH was seen in the group treated with oFF alone. The addition of GnRH antagonist appeared to decrease the rebound, suggesting that this rebound release of FSH may have a GnRH-dependent component. Inhibin concentrations in both oFF-treated groups increased after oFF injection and then declined to pretreatment values. However, a second rise in inhibin concentration, concomitant with the FSH rebound in ewes receiving oFF alone, was seen in the group treated with oFF and GnRH antagonist. As this rise in endogenous inhibin concentration could also act to suppress the rebound release of FSH, it cannot be conclusively proved from this study that GnRH input is required for the generation of the rebound release of FSH after treatment with oFF. PMID- 8410830 TI - Induction of ovulation and superovulation in mares using equine LH and FSH separated by hydrophobic interaction chromatography. AB - Pharmacological control of reproduction in mares requires the use of equine gonadotrophins to avoid induced immunological resistance. Crude equine gonadotrophins (CEG) have been used but the presence of equine luteinizing hormone (eLH) and follicle-stimulating hormone (eFSH) in CEG has led to disappointing results in superovulation studies. Separation of eLH and eFSH activities from CEG is necessary to overcome this problem. The hydrophobic properties of the two hormones were sufficiently different to permit their separation by hydrophobic interaction chromatography (HIC) on a phenyl Sepharose matrix. Good yields of separate FSH and LH fractions were readily obtained by stepwise elution and the method was adapted for large scale preparations of enriched fractions of eLH and eFSH. Two experiments were performed in vivo to evaluate the biological activity of the HIC fractions. Experiment 1 showed that biological activity of the LH fraction in inducing ovulation of preovulatory follicles was similar to that obtained with CEG, indicating that LH bioactivity was not altered by HIC. Experiment 2 demonstrated that biological activity of the FSH fraction was identical (as far as rate of ovulation was concerned) to that of CEG in superovulating mares, indicating that FSH activity was also not altered by HIC. Although we have not obtained better results with the separate equine gonadotrophins than with CEG, it is potentially advantageous to use preparations with single activity to obtain a controlled balance of FSH and LH activity. The HIC technique was chosen because it could easily be scaled up to provide the large amounts of the separate hormones needed for the treatment of a large number of mares. PMID- 8410831 TI - Use of primary cultures of rabbit oviduct epithelial cells to study the ionic basis of tubal fluid formation. AB - A pure population of rabbit oviduct epithelial cells was isolated and grown as a polarized monolayer on collagen-impregnated filters in primary culture. The cells were shown to be epithelial by immunocytochemical staining. The cells were mounted in a modified Ussing chamber which enabled ion transport across the cells to be studied. There was a net flux of Cl- in a basal to apical direction which was reversed by 1 mmol dibutyryl cyclic AMP l-1 (cAMP). A small but consistent transepithelial electrical potential difference (p.d.) of 0.86 mV was recorded with the apical side of the cells negative with respect to the basal. Adrenaline added to the basal side of the cells induced large transient increases in p.d. across the monolayer, involving both alpha and beta receptors. Adrenaline also induced a small increase in basal to apical Cl- transport across the cells. It is proposed that adrenergic agonists and cAMP modulate rabbit oviduct fluid formation in part via an effect on transepithelial chloride transport. PMID- 8410832 TI - Development of an LH receptor assay capable of measuring serum LH/CG in a wide variety of species. AB - The development of a radioreceptor assay (RRA) that can measure serum LH in a variety of species and CG in sera and urine of pregnant women and monkeys is reported. Using sheep luteal membrane as the receptor source and 125I-labeled hLH/hCG as the tracer, dose-response (displacement) curves were obtained using hLH or hCG as standard. The addition of LH-free serum (200 microliters per tube) had no affect on the standard displacement curve. The assay is simple, requires less than 90 min to complete and provides reproducible results. The sensitivity of the assay was 0.6 ng hLH per tube and the intra- and interassay variations were 9.6 and 9.8, respectively. Sera obtained from male and female bonnet monkeys (Macaca radiata) and monkey pituitary extract showed parallelism to the standard curve. The concentrations of LH measured correlated with the physiological status of the animals. Sera of rats, rabbits, hamsters, guinea-pigs, sheep and humans showed parallelism to the hLH standard curve indicating the viability of the RRA to measure serum LH of different species. Since the receptors recognize LH and CG, detection of pregnancy in monkeys and women was possible using this assay. The sensitivity of the assay of hCG was 8.7 miu per tube. This RRA could be a convenient alternative to the Leydig cell bioassay for obtaining the LH bioactivity profile of sera and biological fluids. PMID- 8410833 TI - Reproductive wastage in the androstenedione-immune ewe. AB - An experiment was carried out using 320 adult Merino ewes to examine the effects of immunization against an androstenedione human serum albumin conjugate (Fecundin) on ovulation rate, fertilization rate and embryo viability at days 2, 9 and 13-14 after fertilization. The ovulation rate of immunized ewes (2.19 +/- 0.06) was greater (P < 0.001) than that of control ewes (1.43 +/- 0.04). The recovery rate of embryos or of unfertilized oocytes on day 2 was reduced in immunized ewes, but fertilization rate of recovered oocytes was unaffected by immunization. The mean number of normal embryos per ewe pregnant (prolificacy) was higher and the proportion of ewes pregnant (fertility) was lower in immunized than in the control ewes. The distribution of the number of cells per embryo showed no differences in developmental age over the period of sampling, the majority of embryos at this time being at the two- to four-cell stage of development. At day 9 of pregnancy, blastocyst recovery rates were lower in immunized than in control ewes. About 90% of blastocysts recovered were developing normally in control ewes compared with 64% in immunized ewes. The majority of blastocysts recovered on day 9 had hatched from the zona pellucida prior to recovery (mean values were 94.2% and 87.8% for control and immunized groups, respectively). In control ewes single blastocysts were larger than twin blastocysts, but for the immunized ewes this difference was not significant. Both single blastocysts (P < 0.01) and twin blastocysts (P < 0.05) from immunized ewes were smaller than those from control ewes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410834 TI - Evidence for a circannual rhythm of reproduction and prolactin secretion in a seasonally breeding macropodid marsupial, the Bennett's wallaby (Macropus rufogriseus rufogriseus). AB - Two groups of adult female wallabies were maintained in photoperiod-controlled rooms from June 1987 until August 1988. Group SSH was held on summer solstice photoperiods throughout the experiment; group SN was subjected to weekly stepwise simulated natural changes in photoperiod. Plasma melatonin concentrations reflected photoperiod with high concentrations during the dark phase in both groups. Group SN wallabies commenced oestrous cyclicity on 21 July (+/- 19 days, n = 6) entered reproductive quiescence on 14 February (+/- 10 days, n = 5) and recommenced cycling on 8 June (+/- 3 days, n = 4). Group SSH wallabies began cycling on 27 July (+/- 9 days, n = 7) at a time that was not significantly different from that of group SN. Three out of five of group SSH exhibited a spontaneous period of reproductive quiescence of between 59 and 70 days commencing between 3 December and 25 February. There was a highly significant difference between the transient plasma prolactin response to a dopamine antagonist during cycling and quiescent periods in both groups (P < 0.001) such that the response was increased during periods of quiescence. Our data support the hypothesis that prolactin is involved in the control of seasonal quiescence in the female Bennett's wallaby and demonstrate that spontaneous changes in reproductive state and prolactin can occur when animals are maintained on unchanging long photoperiods. PMID- 8410835 TI - Aromatase activity and oestrogen concentrations in bovine cotyledons and caruncles during gestation and parturition. AB - Two peaks of aromatase activities were detected in bovine cotyledons, 2.76 +/- 0.96 pmol min-1 mg-1 protein at month five of gestation and 3.62 +/- 0.78 pmol min-1 mg-1 protein immediately after parturition, but aromatase activity in caruncles was very low throughout gestation. Oestrone concentrations in the cotyledons and caruncles were 0.93 +/- 0.50 and 1.51 +/- 0.69 ng mg-1 protein at month five of gestation, but 4.31 +/- 1.66 and 3.36 +/- 0.98 ng mg-1 protein immediately after parturition, indicating a biphasic pattern. Our findings suggest that oestrogen synthesis in the bovine placenta increased to a maximum at least twice during the period from gestation to parturition; at month five of gestation and at parturition the changes in aromatase activity may only be partially explained by changes in oestrogen production. PMID- 8410836 TI - Germ-line chimaeras can produce both strains of fowl with high efficiency after partial sterilization. AB - The drug busulphan is known to be cytotoxic to migrating primordial germ cells (PGCs). A technique is described in which doses of 0, 25, 50 and 250 micrograms busulphan in 40 microliters sesame oil were injected into the yolk of White Leghorn eggs incubated for 0, 24, 48 and 72 h. The percentage survival values of these embryos showed that the older the embryo at the time of injection, the greater the survival. Increasing the dose of busulphan decreased the survival. The percentage of embryos showing abnormalities increased with higher doses of busulphan. The number of germ cells in histological sections from gonads of 16 day embryos was estimated and in embryos treated with 50 micrograms and 250 micrograms busulphan the number of germ cells was significantly less than in the controls. Eggs were injected with 50 micrograms busulphan at 24-30 h, and at 50 55 h the embryos received an intravascular injection of a germinal crescent cell suspension containing PGCs from Rhode Island Red embryos. Twenty hatchlings from these experiments were raised to sexual maturity. All these birds were fertile and half of the breeding groups producing offspring from the transferred germ cells at a rate of about 35% of the total. The technique would improve the efficiency of producing transgenic gametes. PMID- 8410837 TI - Prolactin as a luteotrophin during late pregnancy in pigs. AB - The effect of prolonged hyperprolactinaemia on the secretion of LH, progesterone and oestradiol, and its relationship to the maintenance of pregnancy was examined in pigs. Twelve crossbred, pregnant gilts were injected i.m. with 1.5 mg haloperidol kg-1 body weight (n = 6) or vehicle (n = 6) once a day from day 60 to day 66 of pregnancy. Blood samples were collected at 08:00, 12:00, 16:00, 20:00, 24:00 h from day 60 to day 67 and every 15 min for 4 h (08:00-12:00 h) on days 60, 63 and 66. Plasma concentrations of prolactin were higher (P < 0.001) in haloperidol-treated gilts than in control gilts (121.3 +/- 4.3 ng ml-1 and 13.6 +/- 0.4 ng ml-1, respectively). Hyperprolactinaemia completely inhibited the pulsatile secretion of LH and diminished (P < 0.001) basal peripheral concentrations of LH (hyperprolactinaemia, 0.3 +/- 0.04 ng ml-1 and control, 0.6 +/- 0.005 ng ml-1). Despite the inhibition of LH release in hyperprolactinaemic gilts, plasma concentrations of progesterone were higher (P < 0.001) than in the control group (20.8 +/- 0.6 and 12.6 +/- 0.2 ng ml-1, respectively). Oestradiol concentrations were not different between groups, although oestradiol tended to be higher in hyperprolactinaemic gilts than in the control group throughout the sampling period (29.1 +/- 1.9 versus 23.7 +/- 1.6 pg ml-1, respectively). Abortion did not occur in any of the gilts. These results are the first to demonstrate that induced hyperprolactinaemia during the second half of pregnancy (days 60-66) will drastically suppress the major porcine luteotrophin but not affect pregnancy maintenance in pigs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410838 TI - The gatekeeper. PMID- 8410839 TI - Rotational handle for laparoscopic instrumentation. AB - A rotational handle was designed to activate various grasping jaws used in operative laparoscopy. The cylindrical handle is in line with the actuating rod operating the jaws. Moving a sliding front post toward a stationary proximal end of the handle opens the self-closing jaws. The instrument serves as a probe while inactive. While one grasps tissues, the handle permits the hand to remain relaxed and to produce smooth manipulations in a full range of 360 degrees. PMID- 8410840 TI - Laparoscopic oophorectomy. AB - Oophorectomies were performed on 312 women as part of an operative laparoscopic procedure over an eight-year period. The average age of the women was 40.4 +/- 0.6 (SEM) years; 36.5% had previously undergone a hysterectomy. The median operating time was 120 minutes. The length of hospitalization was less than 24 hours in 77.6% of women, between 24 and 48 hours in 15.1% and over 48 hours in 7.4%. Intraoperative and/or postoperative complications occurred in 12 women (3.8%). Estimated blood loss greater than 300 mL occurred in two women. The most frequent diagnoses were endometriosis/endometrioma, functional cysts and normal ovarian tissue (usually from ovaries enmeshed in adhesions); two ovaries demonstrated borderline malignant potential. Laparoscopic oophorectomy is acceptable under appropriate conditions. Further studies are necessary to provide criteria for accurate differentiation of benign from malignant ovarian enlargement. PMID- 8410841 TI - Abnormal uterine bleeding associated with iron-deficiency anemia. Etiology and role of hysteroscopy. AB - We reviewed the clinical and histologic records of 61 consecutive premenopausal women with abnormal uterine bleeding and moderate to severe iron-deficiency anemia investigated in a tertiary care and referral center. Excessive bleeding was caused by benign lesions in 67% of the cases and by anovulation in 25% and was unexplained in 8%. Hysteroscopy revealed an organic intrauterine lesion (submucous myomas in 38%, endometrial polyps in 13%, submucous adenomyomas in 3%) that could be treated endoscopically in more than half the patients. In populations without nutritional deficiencies, a woman of reproductive age with sideropenic anemia and no other evident cause of blood loss or systemic disease should be considered menorrhagic until proven otherwise. Hysteroscopy should be included in evaluations of abnormal uterine bleeding. PMID- 8410842 TI - Hysteroscopic management of intractable uterine bleeding. A review of 103 cases. AB - One hundred three patients with intractable uterine bleeding were treated using the continuous flow resectoscope. All patients underwent diagnostic hysteroscopy and endometrial sampling prior to surgery. The patients were divided into three groups. Group I (n = 69) patients had a normal hysteroscopic examination. Group II (n = 26) had intrauterine pathology visible on their diagnostic hysteroscopy. Group III (n = 8) had failed to improve satisfactorily on previous hysteroscopic ablation of the endometrium using the Nd:YAG laser. Group I and III patients underwent hysteroscopic ablation of the endometrium using a 3-mm, ball-end electrode. Group II patients underwent hysteroscopic resection of an intra uterine lesion utilizing a 7-mm wipe loop electrode. The results in all three groups were analyzed and compared. Seventy patients underwent endometrial preparation with one of five pretreatment regimens: danazol, depoleuprolide, depomedroxyprogesterone acetate and surgery. PMID- 8410843 TI - Decreasing the degree of hypothermia during prolonged laparoscopic procedures. AB - During the recent surge of advanced laparoscopic procedures, one operative complication, hypothermia, continues to surface. In a retrospective analysis of patients' temperatures during cases ranging between three and six hours, an average beginning temperature of 36.1 degrees C was lowered to an average of 33.3 degrees C. In the recovery room, an average of four to six hours was necessary to return the patient to a normal temperature using warm blankets and a K-pad. The use of warm fluids (intravenous and irrigating) plus a K-pad under the patient during a three- to six-hour operation reduced the temperature drop an average of 0.9 degrees C with a postoperative return to normal within one hour. The use of the above measures plus a Bair Hugger warmer during the three- to six-hour operations reduced the average temperature decrease to 0.5 degrees C, with a return to normal by arrival in the recovery room. PMID- 8410844 TI - Hysteroscopic treatment of ectopic intrauterine bone. A case report. AB - Hysteroscopic removal of ectopic bone in the uterus, using laparoscopic control and ultrasonographic confirmation, was used to treat a patient who presented with a diagnosis of osseous metaplasia of the uterus. Pathologic analysis revealed benign bony tissue consistent with a diagnosis of osseous metaplasia. Laparoscopy and hysteroscopy confirmed the presence of bone in the form of spicules perpendicular to the uterine endometrium. Most of the bone was present in the posterior portion of the fundus. Initial removal was performed with biopsy forceps followed by gentle curettage. The resectoscope was then introduced to visualize any remaining spicules and remove them by mechanical means with minimal use of electrosurgery. Transvaginal ultrasound assisted in identifying bone and confirming its removal during and after surgery. The hysteroscopic procedure was viewed laparoscopically to reduce the risk of uterine perforation. Dense right adnexal adhesions were also lysed. The patient received conjugated equine estrogens for five weeks post-operatively. Ultrasound showed an intrauterine pregnancy of 5 to 6 weeks plus two small calcifications approximately 1 mm each. The patient delivered a healthy infant and has had no recurrent problems. This case report demonstrates the successful use of multiple diagnostic and treatment modalities in the treatment of ectopic intrauterine bone. PMID- 8410845 TI - Laparoscopic removal of gonads in women with abnormal karyotypes. AB - The objective of this study was to evaluate if laparoscopy can be an acceptable alternative to laparotomy for gonadectomy in women with gonadal dysgenesis (46,XY karyotype or 45,X karyotype with evidence of hyperandrogenism). We retrospectively analyzed five cases of gonadal dysgenesis that were managed by laparoscopic gonadectomy. All five patients underwent laparoscopic gonadectomy performed at a day surgery unit of a university medical center without any complications and were discharged the same day. The average operative time was 72 minutes (range, 35-95), the average blood loss was 20 mL, and the average cost was $4,380. We conclude that laparoscopic gonadectomy is a safe, effective and economical procedure to perform on patients with Y chromosomes or evidence of abnormal gonads. Laparoscopy should be accepted as the treatment of choice in these patients by physicians experienced in pelviscopic surgery. PMID- 8410846 TI - Laparoscopic retropubic colposuspension (Burch procedure). A review of 58 cases. AB - Fifty-eight patients underwent laparoscopic retropubic colposuspension (Burch procedure) for the treatment of genuine urinary stress incontinence. Of these 58 patients, 3 developed postoperative detrusor instability, and another 2 had bladder injuries. The overall complication rate was 8.5%, and the success rate was 94.83%. Our limited experience has shown many advantages of laparoscopic retropubic colposuspension over the traditional abdominal retropubic colposuspension; they include easy access to the space of Retzius, better visibility in the operative field, minimal intraoperative blood loss and postoperative need for pain medication, and shortened hospital stay and recovery period. Most patients were discharged from the hospital within 24-36 hours and resumed normal activities within seven days. Previous major pelvic surgery is not a contraindication to this procedure. Based on our initial experience, laparoscopic retropubic colposuspension appears to be a viable alternative to abdominal retropubic colposuspension. PMID- 8410847 TI - Microbiologic effects of atmospheric conditions used in operative laparoscopy. AB - We tested whether the atmospheric conditions used in operative laparoscopy were responsible for its low infection rate. A plexiglass chamber was used to recreate the atmospheric conditions in an operative laparoscopic procedure with CO2 pneumoperitoneum, including a CO2 atmosphere, gas flow and temperature control. Staphylococcus aureus and Escherichia coli were exposed to this atmosphere and their growth compared to that on control plates exposed to standard culture conditions. The ratio of colony growth on experimental versus control plates was 1.08 +/- 0.19 SD for Staphylococcus and 1.13 +/- 0.66 SD for E coli. We noted no direct inhibitory effect of the pelviscopic atmosphere upon microbiologic growth. The factors responsible for pelviscopy's low infection rate remain obscure. PMID- 8410848 TI - Adhesion formation after endoscopic posterior colpotomy. AB - Twenty-two women who had undergone laparoscopic posterior colpotomy at initial operative laparoscopy and later underwent a second laparoscopic procedure were evaluated for adhesion formation. Fifteen women (68%) had myomata removed, 3 (14%) had a dermoid cystectomy, 1 (5%) had a serous cystadenoma removed, and 3 (14%) who had large endometriomata and severe adhesions underwent salpingo oophorectomy. Although filmy adhesions were noted in nine women, no adhesions were noted in the cul-de-sac. Based on our limited results, it does not appear that tissue removal via laparoscopic colpotomy predisposes reproductive-age women to postoperative adnexal adhesion formation. PMID- 8410849 TI - Laparoscopically assisted vaginal hysterectomy. The initial Nashville, Tennessee, experience. AB - It is now possible to begin a difficult hysterectomy via laparoscopy with or without adnexal removal and then complete the operation vaginally. We report our successful experience with laparoscopically assisted vaginal hysterectomy in 62 of 68 patients. Techniques used for hemostatic separation of the uterus and adnexal pedicles included an automatic laparoscopic stapling device (49 cases), bipolar coagulation with sharp transection (11) and combined techniques (2). Minor complications occurred in four patients. Six patients had their operations converted from laparoscopy to laparotomy because of significant adhesions (three), large fibroids (two) and poor access due to obesity (one). The use of a stapling device required less anesthesia time (1 hour, 57 minutes, vs. 3 hours, 43 minutes), a smaller blood loss (145 vs. 247 mL) and shorter hospital stays (2.53 vs. 2.75 days) than did laparoscopic bipolar coagulation. However, the average hospital costs were greater for disposable automatic stapling devices and trocars when compared to bipolar coagulation techniques ($9,310 vs. $6,227). Postoperative patient satisfaction with the operation was high (98%), with a high rate of symptom resolution (95%). Laparoscopically assisted vaginal hysterectomy is a safe, effective operation in selected cases and may soon become a common alternative to abdominal hysterectomy in certain cases. PMID- 8410850 TI - Microbial invasion of the amniotic cavity during term labor. Prevalence and clinical significance. AB - The purpose of this study was to determine the prevalence and clinical significance of microbial invasion of the amniotic cavity during spontaneous parturition at term. Amniotic fluid was retrieved by transabdominal amniocentesis from 90 women in spontaneous term labor with intact membranes. Fluid was cultured for aerobic and anaerobic bacteria, including Mycoplasma. The prevalence of positive amniotic fluid cultures was 18.8% (17/90). The most common microbial isolates were Ureaplasma urealyticum, Streptococcus agalactiae, Lactobacillus species and Mycoplasma hominis. Clinical signs of chorioamnionitis were present in three patients, and only one of them had a positive amniotic fluid culture. Five patients (5.5%) had puerperal endometritis; three of them had an amniotic fluid culture positive for microorganisms. All neonates were free of clinical signs of infection. The data indicate that microbial invasion of the amniotic cavity occurs frequently during spontaneous labor at term, and it may be both the cause and the consequence of labor. PMID- 8410851 TI - Lupus anticoagulant. Significance in habitual first-trimester abortion. AB - Therapy with steroids and aspirin has been reported to benefit pregnancies in patients with lupus anticoagulant (LA). In this study, habitual first-trimester aborters with LA using steroids and aspirin were compared to a control group of untreated habitual aborters without LA. In habitual aborters with LA, 12 of 24 (50%) pregnancies reached the second trimester as compared to 8 of 22 pregnancies (36%) in the control group. Since the treated group did no better than the control group, LA probably is not a cause of first-trimester abortion. However, once the second trimester is reached, a 50% incidence of growth retardation was found, and 42% of fetuses died in the second or third trimester in treated LA patients. Treatment with steroids and anti-platelet aggregating agents may be necessary despite the attendant risks to prevent those sequelae in the second and third trimesters. There was a 29% live birth rate in treated LA patients; the rate was 36% in control patients. However, this rate was produced only by early intervention, which was unnecessary in the control patients. PMID- 8410852 TI - Identifying and managing ethical conflict in the gynecologist-patient relationship. AB - This paper applies the methods of ethics to the problem of differences between the gynecologist and patient about what is in the patient's interest. We develop a framework for clinical judgment and decision making about the ethical dimensions of the gynecologist-patient relationship. To achieve this goal we first define the fundamental principles of beneficence and respect for autonomy. We then show how these two principles interact in clinical judgment and decision making using common examples from gynecologic practice. We examine factors that influence the relative weight of these principles, including subject matter, probability of net medical benefit, availability of reasonable alternatives and ability of the patient to participate in the informed consent process. PMID- 8410853 TI - Endoscopic bladder repair during total modified laparoscopic hysterectomy. A case report. AB - Endoscopic repair was performed on a bladder laceration during total modified laparoscopic hysterectomy, with the vaginal cuff sutured to the cardinal and uterosacral ligaments. The patient underwent the hysterectomy for chronic pelvic pain. The endoscopic bladder laceration was repaired via laparoscopy, and the patient was discharged from the hospital on the second postoperative day with a catheter connected to a leg bag for drainage. Intravenous pyelogram with full bladder distention, which was performed two weeks after surgery, revealed no abnormality. The patient returned to her daily activities, including sex, within three weeks. PMID- 8410854 TI - Cervical cancer and tuboovarian abscesses. A report of three cases. AB - Tuboovarian abscesses are rarely found in cervical cancer cases. In each of the three cases presented below, the diagnosis of adnexal disease was established at exploratory laparotomy. In one patient following surgery the cancer was downstaged from IIIb to IB. Another patient had undergone external radiotherapy with considerable morbidity prior to surgery. The third patient presented with ruptured tuboovarian abscesses. The cases illustrate the spectrum of problems when these diseases coexist. Exclusion of the presence of tuboovarian abscesses in selected cases by surgery is recommended to optimize treatment of the cancer. PMID- 8410855 TI - Complete androgen insensitivity with persistent mullerian structures. A case report. PMID- 8410856 TI - Operative laparoscopy. American Association of Gynecologic Laparoscopists 1991 membership survey. AB - The American Association of Gynecologic Laparoscopists 1991 membership survey on operative laparoscopy had a total of 878 respondents reporting 56,536 procedures, for a 1.5-fold increase over the number of procedures reported in 1988. The most frequent indication was pelvic pain (58% of procedures); next was infertility (42%). Laparoscopic management of ectopic pregnancy increased over twofold, from 1,914 cases in 1988 to 4,620 in 1991. As compared to the 1988 survey data, the rate of unintended laparotomy to manage hemorrhage, bowel or urinary tract injury may have increased, but the death rate remained low, 1.8 per 100,000 procedures. PMID- 8410857 TI - Operative hysteroscopy. American Association of Gynecologic Laparoscopists 1991 membership survey. AB - The 1991 American Association of Gynecologic Laparoscopists membership survey on operative hysteroscopy had a total of 630 respondents (almost double the 1988 number) who reported performing 17,298 procedures as compared to 7,293 in 1988. Directed biopsy and endometrial ablation were the procedures reported most commonly. Endometrial ablation increased fivefold since 1988, and myomectomy increased fourfold. The majority of operative hysteroscopies were performed for a complaint of abnormal bleeding (73%). The most frequently reported complication was uterine perforation not requiring transfusion (11 per 1,000 procedures). The rate of water intoxication or pulmonary edema dropped from 3.4/1,000 in 1988 to 1.4/1,000 in 1991. However, some serious complications (eight laparotomies for bowel injury, three CO2 embolisms and three deaths) were reported for 1991. PMID- 8410858 TI - Laparoscopic sterilization. American Association of Gynecologic Laparoscopists 1991 membership survey. AB - The 1991 membership survey of the American Association of Gynecologic Laparoscopists indicated more laparoscopic procedures than had been reported in the last survey, in 1988. The respondents reported 30,480 sterilizations, 41,160 diagnostic laparoscopies and 8,015 hysteroscopies. The distribution of sterilization by method of tubal occlusion has remained stable since 1985. Complications from diagnostic laparoscopy remain consistently higher than from sterilization. This year the complication rate for diagnostic procedures was higher than in any other year since 1976 (4.9 per 1,000 cases). One death was reported after sterilization and one after diagnostic laparoscopy. PMID- 8410859 TI - Laparoscopically assisted vaginal hysterectomy as definitive therapy for stage III and IV endometriosis. AB - Operative laparoscopy combined with vaginal hysterectomy and salpingo oophorectomy was used to treat advanced endometriosis in 40 of 46 patients. This treatment plan was abandoned in favor of laparotomy in six patients. Of the 40 patients successfully treated by laparoscopically assisted vaginal hysterectomy, 39 are completely free of symptoms. Major complications, including blood loss requiring transfusion and injury to the ureter or bowel, were sustained at acceptable rates. No serious infections occurred. The mean operating time was 191 minutes. Laparoscopically assisted vaginal hysterectomy and removal of all ovarian tissue combined with excision of all endometriosis may be used as definitive therapy for advanced endometriosis. The major complications associated with surgical therapy for high-stage endometriosis are encountered; therefore, laparoscopic treatment requires advanced laparoscopic surgical skills. PMID- 8410860 TI - Laparoscopic hysterectomy with the Endo GIA 30 stapler. AB - Eighty-two patients underwent laparoscopic hysterectomies from March 1991 to September 1992. The indications were adenomyosis, myomata uteri, intractable menorrhagia and endometriosis with severe pelvic adhesions. The operations were performed through usage of the techniques of videolaparoscopy, including a combination of Kleppinger bipolar forceps for hemostasis and scissors and/or CO2 laser for lysis of adhesions. Fourteen used the Endo GIA 30 stapler in transecting the infundibulopelvic and/or cardinouterosacral ligament. The mean blood loss was 175 mL, and two patients had intraoperative bladder injuries requiring laparotomy repair, but none of them was related to the utilization of the Endo GIA 30 stapler. Most of the patients were discharged on the second postoperative day. The advantages of laparoscopic hysterectomy are short hospitalization, small blood loss and less postoperative discomfort. Implementation of the Endo GIA 30 stapler could facilitate the procedure. PMID- 8410861 TI - Laparoscopic therapy for tubal pregnancy using prostaglandins. AB - Nineteen patients with confirmed tubal pregnancy and constant or rising plasma beta-human chorionic gonadotropin (beta-hCG) levels were treated with laparoscopically guided injection of prostaglandin F2 alpha into the oviduct. Fifteen patients received additional prostaglandin E2 during three consecutive postoperative days. One patient was excluded from the study. The treatment was defined as successful when plasma beta-hCG levels declined below the lower detection limit and no further intervention other than prostaglandin application was required. The treatment was successful in 12 patients (66.7%). Given a beta hCG level greater than 2,500 mU/mL as an exclusion criterion for treatment with prostaglandin, the success rate was 84.6%. Six patients underwent salpingotomy because of rising beta-hCG levels following treatment. The outcome was not related to the postoperative treatment with prostaglandin E2. None of the treated patients displayed any adverse reactions following prostaglandin F2 alpha application. Postoperative hysterosalpingography was performed on six successfully treated patients, demonstrating bilaterally patent fallopian tubes in all of them. Prostaglandin therapy in tubal pregnancy has been proven effective in selected cases. PMID- 8410862 TI - Endometrial ablation. A report of four cases. AB - Three women had evidence of atypical endometrial hyperplasia at the time of endometrial ablation, discovered because they had mechanical preparation of the endometrium rather than medical preparation. In one of these patients, a frozen section revealed atypical endometrial hyperplasia, and the ablation was cancelled. A fourth woman had an unsuccessful endometrial ablation; repeat ablation was followed by nine months of amenorrhea, at which time she began spotting and requested a hysterectomy. The pathology report on the hysterectomy specimen revealed adenomatous hyperplasia. Gynecologists must be vigilant in their evaluation of the endometrium before, during and after endometrial ablation. PMID- 8410863 TI - Bladder and bowel injury after electrodesiccation with Kleppinger bipolar forceps. A clinicopathologic study. AB - To evaluate the extent of electrocoagulation damage in tissue undergoing bipolar desiccation, five mature female pigs were used as models of tissue and adjacent vital structures commonly encountered during operative laparoscopy. Electrocoagulation was performed on bowel mesentery, sidewall peritoneum, ureters, uterus, uterotubal junction and broad ligament; superficial bowel and bladder burns were treated. Kleppinger forceps were also used to coagulate various-sized arteries to determine hemostasis. Histopathologic analysis demonstrated no inflammation or tissue destruction beyond the desiccated area seen immediately after coagulation and no direct correlation between tissue thickness and spread of electrocoagulation. It also appears that blood vessel hemostasis can be achieved safely with Kleppinger forceps on vessels up to 3 mm in diameter. PMID- 8410864 TI - Karyotypic abnormalities and hydramnios. Role of amniocentesis. AB - Hydramnios complicates up to 1.6% of pregnancies, with major fetal malformations found in an average of 20% of patients with hydramnios. Chromosomal abnormalities, although associated with some conditions in which hydramnios is present, have not been reported to complicate a significant percentage of hydramnios cases. Data from 45 patients admitted during the period January 1, 1985-May 31, 1990, were analyzed. All patients had hydramnios sufficiently severe to merit hospitalization for diagnosis and/or treatment. Amniocentesis was performed at the discretion of the attending physician and not under a specific protocol. The incidence of major fetal structural malformations was 36%. Of patients in whom amniocentesis was performed, 22% were found to have karyotypic abnormalities, none of which was specifically suspected prior to the amniocenteses. Neither the estimated gestational age nor the assessment of the amount of amniotic fluid differed between those with karyotypic abnormalities and those with normal chromosome complements. This information suggests that both advanced ultrasound studies and amniocentesis are beneficial in the evaluation of hydramnios. PMID- 8410865 TI - Suppression of pregnancy-induced nausea and vomiting with sensory afferent stimulation. AB - This study examined the effect of sensory affect stimulation (SAS) delivered through the volar surface of the wrist on pregnancy-induced nausea and vomiting. Twenty-three women with significant nausea and vomiting in the first 14 weeks of pregnancy were enrolled in a randomized, crossover study comparing an active SAS unit and an inactive placebo unit. Twenty-one women experienced improvement in symptoms, 20 (87%) with the SAS unit and 10 (43%) with the placebo device. Nine women had an improvement with both devices. Eleven women reported an improvement with SAS only, while one woman had placebo improvement only. SAS applied to the wrist can effectively improve pregnancy-induced nausea and vomiting as compared to a placebo device. PMID- 8410866 TI - Estimation of uterine volume with ultrasound in women with endometrial carcinoma. AB - In calculating uterine volume with ultrasound, it is necessary to calculate the volume of a parallelogram containing the uterus or to assume a specific shape for the uterus. The objective of this study was to determine how much (what percentage) of the parallelogram was actually occupied by the uterus without assuming any specific shape for the uterus. Overall, the volume of the uterus occupies 63% of the volume of the parallelogram. The uterus occupies a greater percentage of smaller parallelogram volumes than larger ones. The data also suggest that the concomitant presence of myomata may be responsible for the positive intercept of the regression line and therefore for the finding that small uteri occupy more than 63% of the parallelogram volume. PMID- 8410867 TI - Wharton's jelly in the umbilical cord. A study of its quantitative variations and clinical correlates. AB - A frozen segment was prepared from each of 398 consecutively collected umbilical cords after stripping them of blood. Three cross-sections were obtained from each cord, and dye imprints were made. Their surface area was measured by placing a transparency with a grid of squares over the imprint and counting the number of squares it occupied. An average of these three cross-sectional areas was taken to estimate the quantity of Wharton's jelly at any point along the umbilical cord. Of the 14 maternal and fetal variables studied, those that were found to be independently related to the estimated quantity of Wharton's jelly in the umbilical cord were male gender (P = < .01), weight of the fetus (P = .05) and maternal prepregnancy weight (P = < .01). These variables, however, explained only 7% of the observed variation in quantity. A further 3.3% was explained by measurement error, leaving 89.7% of the observed variation unexplained. PMID- 8410868 TI - Concordance of 21-hydroxylase gene ratio, human leukocyte antigen haplotyping and adrenal testing results in a family with late-onset adrenal hyperplasia. AB - Late-onset adrenal hyperplasia (LOAH) due to 21-hydroxylase (21-OH) deficiency is one of the most common autosomal recessive disorders. There appear to be two 21 OH genes, CYP21A (a pseudogene) and CYP21B (the functional gene), which lie in close proximity to the human leukocyte antigen (HLA) encoding region on the short arm of chromosome 6. While the CYP21A/CYP21B ratio is normally 1:1, ratio abnormalities are frequent in LOAH, suggesting gene deletion, duplication or conversion. The objective of this study was to determine whether an abnormal CYP21A/CYP21B ratio could predict carriers of LOAH, as determined by endocrine and HLA results. The probands appear to be compound heterozygotes carrying a 21 OH gene for LOAH and a deletion of the homologous gene. However, concordance between an abnormal 21-OH gene ratio and the inheritance of the LOAH gene does not appear to be complete, as demonstrated by this family study. Further studies of the feasibility of screening carriers for 21-OH deficiencies with the CYP21A/CYP21B ratio or other molecular probes must be performed. PMID- 8410869 TI - Successful pregnancy outcome with combination therapy in women with the antiphospholipid antibody syndrome. AB - Four women with the antiphospholipid syndrome associated with lupus anticoagulant and a poor obstetric history were treated with a combination of glucocorticosteroids, anticoagulants and platelet inhibitor therapy. All patients had at least one previous miscarriage while receiving prednisone and low-dose aspirin. The treatment regimen included: aspirin, dipyridamole, prednisone, and warfarin or heparin. This treatment resulted in a successful pregnancy outcome in all cases, without preeclampsia or recurrence of thrombosis. One patient developed a vertebral compression fracture while receiving heparin and prednisone. Two pregnancies required cesarean delivery for fetal distress at 32 and 34 weeks. All four infant birth weights were appropriate for the gestational age. This regimen may be a therapeutic option for patients with the antiphospholipid antibody syndrome, especially if they have failed other commonly used treatments. PMID- 8410870 TI - Association between alcoholic and caffeinated beverages and premenstrual syndrome. AB - We examined the association between alcohol and caffeine consumption and premenstrual syndrome (PMS). One hundred two women with PMS matched by age and race to an equal number of women without PMS were compared for differences in intake of alcoholic and caffeinated beverages, with differences determined using Student's t-tests and conditional logistic regression for matched pairs. Information on alcohol and caffeinated beverage consumption was obtained from three 24-hour dietary recall interviews conducted during the postmenstrual period and from three conducted during the premenstrual period. Results showed that no significant difference was observed in total caffeine intake or in the individual caffeinated beverages consumed during either the post-menstrual or the premenstrual period. For alcohol consumption, however, women with PMS drank 1.41 (95% confidence interval; range, 0.34-2.47) more servings per week during the postmenstrual period. Based on post-menstrual consumption, women in the heaviest drinking category (> or = 10 drinks per week) were significantly more likely to have moderate to severe PMS (P < .005) than nondrinkers. This same significant relation, but to a lesser degree, was observed based on premenstrual consumption. We conclude that because PMS is more strongly associated with alcohol consumed in the symptom-free, postmenstrual period, drinking is unlikely to be simply a response to PMS symptoms as others have previously suggested. PMID- 8410871 TI - Pelvic washings for cytologic analysis in endometrial adenocarcinoma. AB - The adoption of surgical staging into the management of endometrial adenocarcinoma has permitted improved accuracy in the study of prognostic variables in patients with limited disease. In an effort to resolve the current controversy about the importance of pelvic washings for cytologic analysis in endometrial cancer, 270 consecutive patients with neoplasms histologically limited to the uterus were studied. Fourteen patients (5.2%) were found to have positive results. Cytologic results correlated with tumor grade (P < .02) and depth of myometrial penetration (P < .02). Multivariate analysis indicated age 70 years or greater (P < .002), outer-third myometrial invasion (P < .004) and positive washings (P < .02) to predict survival. The probability of five-year survival in the total group was 0.85, which decreased to 0.67 in patients with positive washings. Despite statistical significance as a prognostic variable, the overall clinical utility of peritoneal cytology in planning postoperative adjuvant therapy in this series was limited by its correlation with other prognostic factors and overall low rate of positivity. Conclusions regarding effective adjuvant therapy for positive washings could not be drawn due to the smaller number of patients with abnormal results and the variety of treatments employed. PMID- 8410872 TI - Incarcerated incisional hernia after laparoscopy. A case report. AB - A woman who had undergone operative laparoscopy with myomectomy, appendectomy and coagulation of endometriosis was readmitted on postoperative day 3 with a small bowel obstruction. At laparotomy she was found to have an incarcerated loop of small bowel through a 12-mm trocar site in the left midabdomen. A bowel resection was not required. The defect was closed, and the patient recovered without difficulty. Two points should be made about avoiding hernias, especially through a larger trocar site. First, the trocar sheath should be opened to room air during its removal to avoid creating a vacuum and pulling a loop of bowel into the incision. Second, the fascia should be closed after removal of larger trocars. This can be accomplished during direct visualization through the laparoscope prior to removal of the pneumoperitoneum to avoid placing the suture through the bowel. PMID- 8410873 TI - Henoch-Schonlein purpura associated with eclampsia. A case report. AB - A case of peripartum seizure activity presumed to represent eclampsia is presented. The postpartum diagnosis of Henoch-Schonlein purpura places the original diagnosis in question due to similarities in the manifestations of the two syndromes. PMID- 8410874 TI - Neonatal labioperineal tear from fetal scalp electrode insertion. A case report. AB - Intrauterine electronic fetal heart monitoring is a widely used tool in diagnosing intrauterine fetal distress. We describe a case of successful conservative treatment for a neonatal labioperineal tear caused by scalp electrode application during breech delivery. PMID- 8410875 TI - Chronic retroperitoneal pelvic abscess in the thigh simulating ovarian carcinoma. A case report. AB - A recurrent abscess of the thigh, associated with a broad ligament abscess, presented in a postmenopausal woman with cirrhosis of the liver. This unusual complication has not been reported before. The risk of serious morbidity and mortality from the combination of these two coexisting lesions is obvious, and gynecologists and surgeons should be prepared for early surgical intervention. PMID- 8410876 TI - Treatment of colonic endometriosis with a gonadotropin releasing hormone agonist and pregnancy after human menopausal gonadotropin/intrauterine insemination. A case report. AB - A woman with longstanding primary infertility and progressive, symptomatic rectal endometriosis was treated with daily leuprolide acetate for nine months. All bowel symptoms subsided. The patient was treated with human menopausal gonadotropin and intrauterine insemination prior to discontinuation of the leuprolide acetate, resulting in a twin pregnancy. PMID- 8410877 TI - Diagnosis and treatment of cervical pregnancy. A case report. AB - A case of preoperative diagnosis of cervical pregnancy by transvaginal sonography is presented. The widespread use of transvaginal sonography facilitates earlier detection of this kind of ectopic gestation, thus improving the prospects for conservative treatment with preservation of fertility. PMID- 8410878 TI - Intraarticular hyaluronan injections in the treatment of osteoarthritis: state-of the-art review. AB - Viscosupplementation (restoring the rheological properties of a tissue matrix) by injection of hyaluronan into the joints has been in use for 2 decades, mostly for osteoarthritis (OA) of the knee, using doses of 20-25 mg of hyaluronan of 500,000 to 2,500,000 M(r), in sequences of 2 to 10 weekly injections. Pain relief appears in a few days, progresses over a few weeks, and often lasts several months. Some data suggest the benefit can last 6 months to one year. Tolerance is universally reported as very good. Those responding to hyaluronan are 65-80%, compared to 30 35% responding to control. Compared to local steroid injections, the effect of hyaluronan appears significantly more lasting. More highly viscoelastic preparations of hyaluronan can be expected to make this therapy even more attractive. PMID- 8410879 TI - An analysis of clinical studies of the use of crosslinked hyaluronan, hylan, in the treatment of osteoarthritis. AB - Viscosupplementation with hyaluronan is a safe modality for treating osteoarthritis (OA) of the knee, but it often requires many injections to achieve efficacy. This may result from insufficient elastoviscous properties of current hyaluronan preparations or too rapid egress from the joint. Because both of these properties would be improved by increasing the molecular weight of the substance, hylans, or crosslinked hyaluronans, were developed as therapeutic agents for viscosupplementation of osteoarthritic synovial fluid. This report reviews the results of 4 clinical trials in Germany that assessed the efficacy and safety of hylan G-F 20 (Synvisc). PMID- 8410880 TI - The pathophysiology of osteoarthritis and the implication of the use of hyaluronan and hylan as therapeutic agents in viscosupplementation. AB - Osteoarthritis (OA) is characterized by several pathological events, including a progressive erosion of the articular cartilage (particularly the weight bearing areas of the joint), synovial inflammation, which may contribute to disease progression, and changes in the lubricating properties of the synovial fluid (SF). We discuss the pathological features that may be involved in the progression of OA, and the implications of the restoration of the viscoelastic properties of the SF (viscosupplementation) as a therapeutic modality for the treatment of OA. PMID- 8410881 TI - Viscosupplementation: a new concept in the treatment of osteoarthritis. AB - Viscosupplementation is a new medical concept that has as its therapeutic goal the restoration of rheological homeostasis in pathological structures such as osteoarthritic joints. When the normal viscoelasticity of a solid tissue compartment or the elastoviscosity of a liquid tissue compartment is decreased under pathological conditions, normal function and regenerative processes are impaired. By introducing viscosupplementary devices, the normal rheological state of such compartments is restored or augmented. These devices stay in the tissue compartment for various periods of time, depending on the nature of the viscosupplement and the pathophysiology of the tissue compartment. PMID- 8410882 TI - Medicine and post-modernity. PMID- 8410883 TI - Surgery: science and craft. PMID- 8410884 TI - Abnormal glycine metabolism in motor neurone disease: studies on plasma and cerebrospinal fluid. AB - Plasma amino acid levels were measured following oral glycine loading in 43 patients with motor neurone disease (MND), eight normal subjects and 18 neurological disease controls with wasting or spasticity from a variety of other causes. Levels at baseline and 1.5 h after loading did not differ, but at 4 h, plasma glycine levels in MND patients remained significantly higher than in normal and neurological controls (P < 0.013). Cerebrospinal fluid glycine levels, which were maximal at 2.5 h, were also significantly higher in MND patients than neurological controls (P < 0.04). These observations suggest a defect of glycine 'housekeeping' in the central nervous system in MND which may be relevant to the pathogenesis of the disease. PMID- 8410885 TI - Percutaneous antegrade ureteric stent insertion in malignant disease. AB - Untreated, progressive bilateral ureteric obstruction, or unilateral ureteric obstruction in patients with a solitary functioning kidney, will ultimately lead to uraemia, renal failure and death. This paper describes 34 successful percutaneous antegrade ureteric stent placements from 37 attempts, performed on 25 selected patients with a history of malignant disease, in whom retrograde ureteric stenting was impossible or difficult. PMID- 8410886 TI - Topical guanethidine relieves dentinal hypersensitivity and pain. AB - The topical application of guanethidine solution to the affected teeth has been successful in relieving natural and clinically induced dentinal pain and hypersensitivity in 13 of 14 dental patients in a pilot study. The mechanism of this analgesia was unclear but probably involved anti-noradrenergic block as there was no evidence of local anaesthesia. Further studies may generate new ideas about the cause of the excruciating pain and hypersensitivity often associated with the acute exposure of tooth dentine and lead to new ways of managing the discomforts of dental conservation and restoration. PMID- 8410887 TI - Psychological aspects of recurrent chest pain. AB - Of 113 consecutive patients admitted recurrently with chest pain 28% exhibited psychiatric morbidity using the GHQ-28 (General Health Questionnaire). Eighty patients had ischaemic heart disease, 17 had non-specific pain and 13 were excluded because of other organic causes for their pain. Of the non-specific group, seven had been admitted previously with chest pain. In common with previous studies of first admissions with non-specific chest pain, recurrent admissions were younger and predominantly male. They also had a history of greater alcohol and cigarette use than patients with ischaemic heart disease. Greater psychiatric morbidity was not demonstrated in this small group of patients. In spite of the absence of an organic aetiology, patients with non specific pain showed similar rates of re-admission to those with ischaemic heart disease before and after the study. Further research is indicated to identify aetiological and maintaining factors for continued non-specific pain. PMID- 8410888 TI - Sensory loss in syringomyelia: not necessarily dissociated. AB - The pattern of sensory loss was assessed in 210 cases of syringomyelia. Dissociated sensory loss occurred in 49% of cases, indicating that its occurrence is not a necessary finding for the diagnosis of syringomyelia. Syringomyelia should be considered in the differential diagnosis of all cases of spinal cord disease. PMID- 8410889 TI - Complications following general anaesthesia for cataract surgery: a comparison of the laryngeal mask airway with tracheal intubation. AB - The incidence of airway complications following general anaesthesia using either a tracheal tube or a laryngeal mask airway was compared in a prospective, randomized study of 79 patients undergoing elective cataract surgery using a standard anaesthetic technique. Assessment was made both at extubation (which was taken to include removal of the laryngeal mask airway) and for 25 min afterwards. There was a significantly greater incidence of coughing prior to extubation (P < 0.001), at extubation (P < 0.001) and after extubation (P < 0.001) in the tracheal group than in the laryngeal mask airway group. No other airway complications were seen in either group. PMID- 8410890 TI - Moving towards Europe: international clinical conferencing on the telephone. PMID- 8410891 TI - Total hip and knee replacements: a survey of 261 hospitals in England. AB - A survey of all 261 National Health Service hospitals in the 14 regions of England performing joint replacement surgery was carried out to determine inter regional variations in the surgeon's choice of hip and knee replacements. There are over 30 types of cemented, 35 types of uncemented hip replacements, and more than 35 types of knee prostheses for the surgeon to choose from. The Charnley low friction arthroplasty is the market leader in hip surgery and is used in 193 hospitals (74%). There is a marked North-South divide in the use of this prosthesis. The Kinematic and Insall Burstein prostheses are the most popular knee replacements. At present, there is a huge choice of joint replacement on the market and many implants have yet to prove that they offer any real advantages over established and often cheaper alternatives. It is suggested that special centres for joint replacement surgery within each region should run clinical trials to determine which prostheses have the best long-term results. The resulting standardization of implants within each hospital would enable Regional Health Authorities to reduce costs in the short-term by cutting stock levels, and should reduce costs in the long-term by reducing the revision rate of failed joint replacements. PMID- 8410892 TI - An information booklet for employers on inflammatory bowel disease: an evaluation by patients. AB - Forty-eight patients with Crohn's disease were offered an educational booklet about inflammatory bowel disease directed at employers. They were asked to assess its value and the best method of distribution. Forty-eight per cent agreed to take part in the study and, of these, 74% completed the project. Although the book was considered valuable, and most patients believed it should be made available to their line managers, distribution was considered best left in the hands of doctors and self-help groups. PMID- 8410893 TI - Tracing patients from acute psychiatric wards. AB - A random sample of those admitted to acute psychiatric wards in Sheffield in 1985 was traced to establish whether or not the patients were homeless 5 years later. Contrary to expectations none were found to be homeless. Although the proportion of mentally ill amongst the homeless may be significantly high, the number discharged from psychiatric hospital, at least in Sheffield, living consistently 'on the streets' or staying regularly in night shelters seems small as a proportion of all discharges. PMID- 8410894 TI - Whiplash injury and peer copying. AB - Whiplash injury has increased during the past 40 years. Peer copying has occurred both among patients and practitioners. The importance of understanding the common course of the injury is stressed and simpler and less costly management suggested. Also noted is the importance of digesting whole articles. Reliance upon abstracted material alone or parts of articles may prove to be misleading and may lead to perpetuating questionable information. PMID- 8410895 TI - The ideology of the human genome project. PMID- 8410896 TI - Why did Nazi doctors break their 'hippocratic' oaths? PMID- 8410897 TI - The history of portal hypertension. PMID- 8410898 TI - A rare cause of upper gastrointestinal bleeding. PMID- 8410899 TI - Isolated hypoglossal nerve palsy caused by carotid artery aneurysm. PMID- 8410900 TI - Cutaneous extravascular necrotizing granuloma (Churg-Strauss) as a paraneoplastic manifestation of non-Hodgkin's B-cell lymphoma. PMID- 8410901 TI - Synchronous breast cancer with different histology. PMID- 8410902 TI - Genital lichen sclerosus et atrophicus. PMID- 8410903 TI - Utilization of the Helicopter Emergency Medical Service. PMID- 8410904 TI - What's in a title--Mr or Dr? PMID- 8410905 TI - AIDS--is the battle being lost? PMID- 8410906 TI - Quality improvement and the hospital patient representative. AB - A national survey of hospital patient representatives was conducted. This study sought to identify and analyse basic demographic and occupational data of patient representatives. An analysis of the patient representatives' perceived roles in the hospital is also conducted. A total of 387 randomly selected United States hospitals were surveyed with a response rate of 66% (256 hospitals). We were able to use only 172 'usable' responses. Of those who responded, we found the typical patient representative to be a female aged 30-50 years old, married with a college degree, and about 6 years of experience from a northeast hospital. In general, the majority of the respondents were very positive of their role in the liaison activity between the patient and the administration. PMID- 8410907 TI - Fathers and breast feeding: a pilot observational study. AB - A small pilot survey (n = 113) by questionnaire of the fathers of a sample of children under one year of age was undertaken in order to investigate the involvement of fathers with infant feeding and their attitudes to the method of feeding adopted. The response rate was 72% overall and 79% when the partners of 'single parent' mothers were excluded. Nearly 30% of respondents had not discussed the method of feeding with anyone, but over 60% had discussed it with their partner. 64% of fathers sometimes helped with feeding their child and 17% said that they always helped. The majority of fathers did not mind their partner breast feeding in front of friends or relatives but 42% did not like them feeding in front of strangers and over half did not like them breast feeding in a public place. From this study, based on relatively small numbers, we conclude that fathers may feel left out of infant feeding. They should be given more opportunity to become involved from an early stage and take part in the decision about the method of infant feeding to be adopted. PMID- 8410908 TI - Longer term functional outcome and societal implications of upper limb fractures in the elderly. AB - To ascertain how they cope with upper limb fractures and the implications for health and social services (H&SS) a cohort of elderly patients was followed up. 42 of the original 51 (82%) patients were interviewed almost one and a half years after sustaining their fracture. Information was collected on ability to perform activities of daily living (ADL), general health, and use of H&SS help from informal carers. Although there was an overall improvement in the ability to perform ADL, many (26%) patients reported a residual difficulty. Informal carers provided most of the support with some contribution from the statutory agencies. Demographic changes with increasing numbers of elderly and increasing employment of married women--the traditional carers, and epidemiological changes with increasing age-specific incidence of upper limb fractures mean that H&SS provision in the future will need to be increased. PMID- 8410909 TI - Insecticides against headlice in Glasgow. AB - A postal questionnaire for describing current practices of insecticide usage for the prevention and treatment of pediculosis was sent to 53 pharmacists in Glasgow. 91% returned completed questionnaires. Between 19,000 to 36,000 bottles of insecticide against headlice were bought by the public in Glasgow in 1991. Most of these were sold in small volumes (less than 100 ml) and sales were highest during the autumn. Although pharmacists sold a range of different classes of insecticide, the most popular were those that contained malathion, the treatment for pediculosis recommended by the Health Board. Choice of treatment was probably influenced by advice given to the public by pharmacists and general practitioners. Clients preferred shampoo formulations. There was evidence that treatments were used prophylactically against headlice. However, there was little indication of large scale resistance to insecticides in the louse population. The results indicate that headlice remain a persistent problem in Glasgow, despite the public adhering to the advice of health professionals. PMID- 8410910 TI - Factors associated with awareness and knowledge of cervical cancer in a community: implication for health education programmes in developing countries. AB - An attempt has been made to study the correlates of knowledge of cervical cancer in a community. The survey undertaken is a part of Knowledge, Attitude and Practice (KAP) study prior to initiation of cytological screening. Total women interviewed by KAP survey were 1411 selected through 2 stage stratified random sampling. Subjects for the present analysis consisted of a group of women who had reported previous gynaecological problems related to cervical cancer. The study brought out that younger women had better awareness and knowledge about cervical cancer and related information. Literacy status for education and exposure to family planning was influential in creating awareness about cervical cancer. Lastly, the earlier episodes of gynaecological problems, and treatment seeking behaviour lead to higher awareness (efforts may be made to innovate ways to reach older and illiterate women at risk of cervical cancer for better awareness in the community. PMID- 8410911 TI - Hull healthline: a description of a telephone health information and promotion service. AB - The development and use of a telephone health information service, Hull Healthline, is described. The local circumstances which make the Hull service different from, and it is argued more effective than, national schemes are detailed. The relevance of such a service to a health promotion strategy is discussed. PMID- 8410912 TI - Exploration of the frontiers of tradomedical practices: basis for development of alternative medical healthcare services in developing countries. AB - The study is a brief exploration of the functions and roles of the traditional healers in the total health care delivery system as a basis for tapping the salient features of this age old art: for the purpose of refining, and establishing it as an alternative medical health-care service. The investigation is considered relevant particularly in the developing countries where, in addition to the dearth of orthodox medical services, institutions and personnel, it is relatively cheaper, socio-culturally accessible and acceptable. Refining and developing some aspects of the traditional healers' services will serve the interest of the health consumers whose main concern is with service and not the source. Furthermore, it is hoped that the study will stimulate purposeful discussions on the need for an unbiased examination of the materials, methods and techniques of the traditional healers including, eventually, compiling a native pharmacopoeia. A more comprehensive account of the traditional healers contributions to the battle against diseases and maintenance of health and well being is envisaged. PMID- 8410913 TI - Health and personal services since 1979: the new managerialism. PMID- 8410914 TI - Occupational health: control and monitoring of psychosocial and organisational hazards at work. AB - This paper considers the conceptualizations which underpin the traditional approach to occupational health and offers a critique; much of which points up the importance of psychosocial and organizational issues. It then explores the possible development of a new 3 factor model of concern - organisational health - based on health, work and organisation. It takes as its thesis the need to address the problems posed by psychosocial and organisational hazards at work and the argument that these can be managed effectively within the framework of practice that has proved successful for the more tangible workplace hazards. PMID- 8410915 TI - Sex and the nation's health. AB - The Government's 'Health of the Nation', influenced by the WHO's 'Global Strategy for Health for All by the Year 2000', pledges a reduction in sexually transmitted diseases in general and AIDS in particular. The recommendations are clearly sensible but how do you persuade people to believe and comply with the 'sensible' advice? Failure to consider public health measures involving sexual behaviour without considering how different cultures respond and interpret the advice given is bound to fail. People are individuals and there is a danger in assuming that our values and beliefs are shared by people globally. For a message to succeed it needs to be simple and direct, and portrayed in its cultural context through an appropriate medium. An appreciation that the message however sensible may clash with individual needs (eg the economics of prostitution) and research into these aspects of sexual behaviour will ensure that the message remains effective. Changing the slogans and methods of conveying the message regularly will help to reinforce key points and reduce apathy. PMID- 8410916 TI - Food hygiene training: the next step. PMID- 8410917 TI - Research-based teaching: a model for teaching environmental health protection issues. PMID- 8410918 TI - Sharing information between health, social services and education re: children with special needs--the issues. PMID- 8410919 TI - Accident prevention and the health of the nation targets. PMID- 8410920 TI - Public health in the United States. PMID- 8410921 TI - Short-term variation in blood pressure determines the accuracy of simultaneous and sequential recordings in the validation of blood pressure measuring devices. AB - Short-term intradevice variation of BP was determined by means of consecutive BP recordings. The intradevice variation was mainly the result of spontaneous, random variation in BP and was the same for invasive, automated oscillometric and auscultatory random zero meter recordings. We show that the accuracy of simultaneous and sequential recordings, performed according to the recommendation of the British Hypertension Society's protocol for the evaluation of automated BP devices, can be calculated by means of statistical analysis if the intradevice variation of the BP is known. We found, for both sequential and for simultaneous recordings, a close agreement between the expected and the observed proportion of differences that fall within certain specified limits and that it is more favourable for a test device to be evaluated by sequential than by simultaneous recordings. PMID- 8410922 TI - Plasma endothelin in essential hypertension and diabetes mellitus. AB - This study was designed to determine plasma endothelin-1 levels in patients with essential hypertension and diabetes mellitus. Endothelin immunoreactivity was measured in normal controls (n = 30), mild-moderate essential hypertensives (n = 25), Type II diabetic normotensives (n = 25) and hypertensive patients (n = 20). In addition, in ten patients of each group we investigated the relationships of endothelin with other vasoactive hormones. Plasma endothelin concentrations were similar in healthy controls, in essential hypertensives and in diabetic patients with or without hypertension, averaging 8.23 +/- 1.68 pg/ml, 7.7 +/- 1.1 pg/ml, 5.05 +/- 0.94 pg/ml, 7.88 +/- 1.41 pg/ml, respectively. No correlations were observed between endothelin and concentrations of plasma renin activity, aldosterone, catecholamines, atrial natriuretic peptide and arginine-vasopressin. The present study suggests that Type II diabetic patients with or without essential hypertension do not have demonstrably higher values of plasma endothelin than essential hypertensives or healthy subjects. PMID- 8410923 TI - Placebo-controlled trial of lisinopril in normotensive diabetic patients with incipient nephropathy. AB - To study the effects of an angiotensin converting enzyme inhibitor, lisinopril, on renal function in incipient diabetic nephropathy, a prospective double-blind randomised placebo-controlled single centre study was set up at our outpatient diabetic-renal clinic. There were 27 patients with Type I and Type II diabetes with an albumin excretion rate of between 20 micrograms/min and 200 micrograms/min, respectively and no hypertension. Intervention treatment with placebo or low dose lisinopril was for 48 weeks. The main outcome changes were in urinary albumin excretion rate, urinary prostaglandin excretion, and glomerular filtration rate. Secondary outcome measures included changes in BP and heart rate. Of the 32 patients entered into the study, 27 completed 48 weeks treatment (12 lisinopril, 15 placebo). Mean (+/- SD) urinary albumin excretion rate fell from 57.6 (25.7) micrograms/min (n = 15) at visit 1 to 26.8 (26.7) micrograms/min (n = 12) at visit 7 after 48 weeks treatment in the lisinopril group but not in the placebo group: 119.2 (116.6) micrograms/min (n = 17) vs. 113.7 (77.0) micrograms/min (n = 15). There was a least squares mean treatment difference of 67.6 micrograms/min (95% confidence interval (CI), -115.0 to -20.2, P < 0.01) in favour of lisinopril compared with placebo. After 48 weeks treatment seven lisinopril treated patients were normoalbuminuric and five were microproteinuric; three placebo treated patients were normoalbuminuric, nine were microalbuminuric and three were macroproteinuric. Excretion of prostaglandin-F1 alpha (PGF1 alpha) and thromboxane-B2 (TXB2) fell in the lisinopril treated group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410924 TI - Comparison of the effects of an angiotensin converting enzyme inhibitor and a calcium antagonist in hypertensive, macroproteinuric diabetic patients: a randomised double-blind study. AB - The objective of this study was to compare the effects of an angiotensin converting enzyme inhibitor, lisinopril, with those of a calcium blocker, nifedipine, on BP control and renal function, in a prospective randomised double blind, double-dummy trial lasting 19 weeks in patients with diabetic nephropathy. We enrolled 28 diabetic patients with hypertension and macroproteinuria from the out-patient diabetic-renal clinic. The antihypertensive treatment consisted of lisinopril or nifedipine, and their effect on arterial BP, urinary albumin excretion, glomerular filtration rate, and renal blood flow were measured. BPs at entry were 166/99 (SD 23/9) mmHg for the lisinopril group and 165/99(21/7) mmHg for the nifedipine group. BPs fell to 143/88 (17/13) mmHg for the lisinopril group and 148/85(25/10) mmHg for the nifedipine group at the end of the study. The albumin excretion rate fell in the lisinopril group from 738.7 (635.2) micrograms/min to 644.6 (965.2) micrograms/min and rose in the nifedipine group from 981.2 (1022.2) micrograms/min to 1072.5 (908.5) micrograms/min (P = NS). Glomerular filtration rates fell from 105.2 (57.5) ml/min per 1.73 m2 to 72.1 (39.4) ml/min per 1.73 m2 in the lisinopril group and from 109.9 (50.0) ml/min per 1.73 m2 to 82.9 (53.9) ml/min per 1.73 m2 in the nifedipine treated group. Renal blood flow fell from 446.8 (217.9) ml/min per 1.73 m2 to 435.1 (243.3) ml/min per 1.73 m2 for the lisinopril group and from 473.0 (216.4) ml/min per 1.73 m2 to 419.0 (278.6) ml/min per 1.73 m2 for the nifedipine group. Differences between the groups were not significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410925 TI - Meta-iodobenzylguanidine (MIBG) scanning in the diagnosis of phaeochromocytoma. AB - During the period 1984-90, meta-iodobenzylguanidine (MIBG) scans were performed in 23 patients with suspected phaeochromocytoma seen at the Hammersmith Hospital. Sixteen patients had a histologically proven phaeochromocytoma and in 14 of these patients the tumour was demonstrated by abnormal uptake of MIBG. Seven patients did not have a phaeochromocytoma and in all of these the MIBG scan was negative. These findings gave the procedure a sensitivity of 87.5% with a specificity of 100%; positive and negative predictive values were 100% and 77.7%, respectively. MIBG scanning is an extremely valuable technique in the management of patients with suspected phaeochromocytoma but is best employed to localise a tumour which has been confirmed biochemically. PMID- 8410926 TI - Imaging of aldosterone-secreting adenomas: a prospective comparison of computed tomography and magnetic resonance imaging in 27 patients with suspected primary aldosteronism. AB - We have compared prospectively the sensitivity, specificity and accuracy of CT and MRI in a series of 27 consecutive patients (age 23-76 yrs, 17 females, 10 males) with clinically suspected primary aldosteronism. We found 13 patients with a unilateral aldosterone-producing adenoma (11 on the left and 2 on the right side), 6 with idiopathic hyper-aldosteronism and 8 with primary hypertension, which in two cases was associated with a nonfunctioning adrenal adenoma. The diagnosis of aldosterone-producing adenoma was confirmed at surgery and pathology in all cases. Idiopathic hyper-aldosteronism was diagnosed on the basis of the results of dexamethasone-suppressed adrenal scintigraphy and/or selective adrenal vein sampling. MRI correctly identified all cases of aldosterone-producing adenoma, but gave false positive results in five cases: one had idiopathic hyper aldosteronism with bilateral nodular hyperplasia and four primary hypertension, which in two patients was associated with a nonfunctioning adrenal adenoma. Therefore, the sensitivity of MRI was 100%, its specificity 64% and overall diagnostic accuracy 81%. In comparison, CT correctly recognized only eight of the 13 patients with aldosterone-producing adenoma and gave false positive results in three primary hypertensives, including the two patients with a nonfunctioning adrenal adenoma. Therefore, its sensitivity, specificity and accuracy were 62, 77 and 69%, respectively. Based on these results, it could be anticipated that about four of every ten patients with aldosterone-producing adenoma would not be correctly diagnosed by CT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410927 TI - Double-blind comparison between nifedipine and amlodipine for the treatment of essential hypertension. AB - Nifedipine is an effective compound for the treatment of hypertension. However, even as a tablet formulation it is relatively short acting requiring two or three times daily administration. Amlodipine is a long-acting calcium antagonist and effectively lowers BP in patients with essential hypertension. In the present study we compared the BP-lowering effect of nifedipine and amlodipine in patients with essential hypertension. Thirteen patients were studied. They had been on nifedipine tablets for at least four weeks and DBP had been consistently > 95 mmHg. After a further month run-in on nifedipine they entered a randomised double blind crossover study of one month' treatment with either nifedipine tablet (20 mg twice daily) or amlodipine (5 mg once daily). BP was measured 12 and 2 hours after the last dose of nifedipine and 24 and 2 hours after the last dose of amlodipine. There was a significant peak/trough effect while on nifedipine tablets, the BP being significantly higher at 12 hours than at 2 hours after the last dose (155.2/90.9 +/- 4.6/1.7 vs. 136.1/84.8 +/- 4.3/1.7 mmHg; P < 0.001/P < 0.005). There was no overall difference in BP between nifedipine and amlodipine treatment when BPs were taken at the respective troughs (i.e. 12 hours and 24 hours). If anything, amlodipine tended to be slightly more effective at least on supine SBP (155.2/90.9 +/- 4.6/1.7 vs. 147.6/89.1 +2- 4.3/1.8 mmHg; P < 0.05, NS). In conclusion, amlodipine given once daily is at least as effective as nifedipine tablets given twice daily in patients with essential hypertension. PMID- 8410928 TI - Effect of intravenous ketanserin on arterial and intracranial pressures in patients with systemic hypertension following intracerebral haemorrhage. AB - The response of arterial BP and intracranial pressure to an intravenous injection of the serotonin antagonist, ketanserin, was recorded in ten patients with hypertension following intracerebral haemorrhage. Five minutes after ketanserin administration, arterial pressure had fallen by 40/21 mmHg from a baseline level of 213/113 mmHg, and remained below pre-treatment levels for two hours. Intracranial pressure was steady throughout the two hour observation period. Although calculated cerebral perfusion pressures declined and whereas it is not known if cerebral perfusion might have been compromised in zones of borderline ischaemia, the present data suggest that ketanserin might be preferred to drugs known to raise intracranial pressure or agents requiring infusion and intensive haemodynamic monitoring for patients with severe hypertension complicating intracerebral haemorrhage. PMID- 8410929 TI - Role of vasoactive peptides in blood pressure control. PMID- 8410930 TI - Lipid-derived second-messengers in smooth muscle. PMID- 8410931 TI - Causes of essential hypertension: can it be cured? PMID- 8410932 TI - Essential hypertension: a disease with origins in childhood? PMID- 8410933 TI - L-arginine:nitric oxide pathway could play a role in the intracellular regulation of renin release. PMID- 8410934 TI - Structural changes in the heart and carotid arteries associated with hypertension in humans. AB - Hypertension is associated with structural changes in the vascular system and in the heart. This study has examined the relationships between carotid artery intima-media thickness and other risk factors in 52 untreated patients (20 hypertensive). Carotid intima-media thickness was measured bilaterally using a Duplex doppler ultrasonic scanner. In the hypertensive individuals the left ventricle was examined by echo-cardiography and the left ventricular mass index determined. There was a significant association between age and IMT, and both SBP and DBP and IMT. The IMT in the hypertensive group was significantly larger than in the normotensive group and in the hypertensive subjects there was a positive association between left ventricular mass index and IMT. There was no significant difference in calculated media stress between the normotensive and hypertensive groups, probably due to a small increase in carotid intima-media area combined with a small reduction in carotid lumen diameter. Hypertension is associated with a thickening of the intima-media of the carotid artery and an increase in left ventricular mass. Whether these changes in cardiac and arterial structure are in response to similar influences remains to be established. PMID- 8410935 TI - Effect of nifedipine and glyceryl trinitrate on retinal blood flow in normal subjects. AB - The retinal vasculature is an accessible region of the microcirculation in humans which undergoes change in essential hypertension. In this study we have examined the effect of nifedipine and glyceryl trinitrate (GTN) on retinal blood flow velocities. Retinal blood flow was measured in 11 subjects using colour Doppler ultrasound. Nifedipine (10 mg orally) or glyceryl trinitrate (0.3 mg, GTN) was given after 30 minutes recumbancy. Systolic retinal arterial blood flow velocity (SRV), diastolic retinal arterial blood flow velocity (DRV), retinal resistive index (RI), SBP, DBP were measured at -10, 0, 5, 10, 15, 20 and 25 minutes following drug administration. Nifedipine significantly increased SRV (peak effect at 15 minutes) but by 25 minutes SRV did not differ significantly from pre drug values. Nifedipine did not significantly affect DRV, RI, SBP or DBP. GTN significantly increased both SRV and DRV, although both tended to fall towards pre-drug values by 25 minutes. RI, SBP nor DBP were not significantly altered by GTN. Both nifedipine and GTN transiently increase blood flow velocity in the central retinal artery although at the doses used neither agent affected BP or RI. Doppler ultrasound measurement of retinal arterial blood velocity may be a useful technique in the investigation of the retinal microcirculation in essential hypertension. PMID- 8410936 TI - Double-blind trial of vitamin C in elderly hypertensives. PMID- 8410937 TI - Expression of insulin-like growth factor-1 by human ventricular explants in culture. AB - There is increasing evidence from animal studies for autocrine and paracrine systems in the heart. As the local role of these peptide systems in the human heart is not clear, we assessed cardiac explants in culture as a model for the study of growth factor expression by the heart in humans. Human right ventricular septal biopsies were maintained as explants for up to 9 days in serum supplemented medium. Insulin-like growth factor-1 (IGF-1) expression was measured by radioimmunoassay and results corrected for tissue protein content. IGF-1 was synthesised by ventricular explants but not secreted into the culture medium. This culture model has the potential to provide insight into the regulation of and response to growth factor expression by the heart in humans. PMID- 8410938 TI - Nitrendipine in older patients with isolated systolic hypertension: second progress report on the SYST-EUR trial. PMID- 8410939 TI - Neospora caninum in aborted twin calves. AB - Neospora caninum was detected in the brains of twin aborted calves by means of a specific immunohistochemical test on formalin-fixed, wax-embedded tissues. Microscopic lesions associated with this infection included multifocal necrosis and gliosis in the brain accompanied by a mononuclear, perivascular inflammatory reaction and meningitis; and severe, diffuse epicarditis and subepicardial myocarditis characterised by infiltration of macrophages, lymphocytes and plasma cells. This report documents the first diagnosis of this parasite as a cause of bovine abortion in South Africa. PMID- 8410940 TI - Medical management of allergic pruritus in the cat, with emphasis on feline atopy. AB - Pruritus associated with allergic skin diseases is an important reason for consultation in feline medicine. The pathophysiology of pruritus is poorly understood in the cat, but histamine and eicosanoids appear to be important mediators. The medical management of allergic pruritus in cats, especially those with atopy, using glucocorticoids, antihistamines, omega-3/omega-6 fatty acid containing products, and combinations thereof is discussed. PMID- 8410941 TI - Breda virus-like particles in calves in South Africa. PMID- 8410942 TI - Pollen grains of grasses in horse faecal analysis. PMID- 8410943 TI - A cryptosporidium sp in an ostrich. PMID- 8410944 TI - The environment: whose responsibility? PMID- 8410945 TI - Pharmacokinetics of rafoxanide in suckling and weaned lambs following oral administration. AB - The pharmacokinetics of rafoxanide administered orally at 15 mg kg-1 were compared in suckling lambs (n = 8), aged 5-8 weeks and weaned lambs (n = 8), aged 21-22 weeks. A significant difference (p < 0.0001) in area under the curve (AUC) and maximum plasma concentrations (Cmax) were observed between the different age groups. The bioavailability, as measured by AUC and Cmax, was 2.5-3 times greater in the suckling lambs compared to the weaned lambs. No significant differences (p > 0.05) in time to maximum plasma concentrations (Tmax), distribution half-life (T1/2 alpha), elimination half-life (T1/2 beta) and mean residence time (MRT) were observed. It was concluded that the pharmacokinetic differences observed between suckling and weaned lambs may result in an increased susceptibility of suckling lambs to the toxic effects of rafoxanide. Lambs younger than 8 weeks, should preferably not be treated with rafoxanide, or the dose must be reduced. PMID- 8410946 TI - Overberg research projects. XV. The efficacy of different anthelmintics against field strains of nematode parasites of sheep in the southern Cape Province. AB - Controlled anthelmintic tests and faecal egg count reduction tests were carried out on natural infections of nematode parasites in sheep on 4 farms in the southern Cape Province. Albendazole, fenbendazole, ivermectin, levamisole, morantel citrate and a combination of albendazole and closantel sodium were tested. Every farm harboured anthelmintic resistant nematode parasites. Adult as well as immature Teladorsagia spp. were resistant to albendazole and fenbendazole, immature Teladorsagia spp. to albendazole/closantel, adult and immature Nematodirus spp. to albendazole/closantel and adult Nematodirus spp. to fenbendazole. PMID- 8410947 TI - Experimental vermeersiekte (Geigeria ornativa O. Hoffm. poisoning) in sheep. I: An evaluation of diagnostic aids and an assessment of the preventive effect of ethoxyquin. AB - Dried milled Geigeria ornativa, which had been stored at room temperature for 4 years, was fed in rations to sheep. The usefulness of contrast radiography of the oesophagus, biopsies of oesophageal and skeletal muscles, electrocardiography, and clinical pathological parameters for diagnosing vermeersiekte were evaluated. Ethoxyquin was evaluated as a preventive agent. All the sheep developed signs of vermeersiekte but regurgitation was observed in only one animal. Contrast radiography and the examination of tissue biopsies were of diagnostic value in 8 out of 13 and 6 out of 6 sheep respectively. Electrocardiography did not aid in antemortal diagnosis of the condition and selected chemical parameters were non specific. The inclusion of ethoxyquin in the diet of experimental animals (n = 3) prior to and during feeding of G. ornativa did not prevent the development of vermeersiekte. This study confirmed that dried stored G. ornativa retains its toxicity and that regurgitation is not a consistent clinical sign in sheep with vermeersiekte. The diagnostic value of contrast radiography of the oesophagus and examination of oesophageal and muscle biopsies is reported for the first time. PMID- 8410948 TI - Experimental vermeersiekte (Geigeria ornativa O. Hoffm. poisoning) in sheep. II: Histological and ultrastructural lesions. AB - The histological and ultrastructural lesions of sheep (n = 5) with experimentally induced vermeersiekte (Geigeria ornativa O. Hoffm. poisoning) were studied. Histological changes in the muscles of the skeletal system, diaphragm and oesophagus comprised hypertrophy and vacuolation of myofibres. In 2 sheep, foci of myocardial degeneration and necrosis were present. No microscopical changes were detected in the central nervous system and in peripheral nerves of the oesophagus and the hind limb. Ultrastructural alterations in the myocardium, semimembranosus muscle and oesophagus were characterised by concurrent myofibrillar degeneration and mitochondrial swelling. The earliest myofibrillar change appeared to be loss of the thick filaments followed by fragmentation and disorganisation and eventually by lysis of central myofibrils. Focal thickening, fragmentation and clumping of Z-band material and dissociation and fragmentation of intercalated discs accompanied the myofibrillar and mitochondrial alterations. PMID- 8410949 TI - Sites of attachment of Karoo paralysis ticks (Ixodes rubicundus) on three cattle breeds. AB - The sites of attachment of female Karoo paralysis ticks (Ixodes rubicundus) were determined on Friesian, Bonsmara and Nguni cattle in the south-western Orange Free State. The largest proportion of the ticks (> 30%) attached to the forelegs and shoulders of the cattle. Contrary to the findings on small stock, no significant differences in the preferred sites of attachment were evident between the breeds of cattle. PMID- 8410950 TI - Besnoitiosis in a horse. AB - Besnoitiosis was confirmed in a pony which presented with inspiratory dyspnoea, scleroderma and ventral oedema. Numerous cysts were visible in the sclerae. Histological examination of the skin confirmed the presence of numerous cysts. The parasite could not be transmitted by subcutaneous injection of homogenised skin from the infected horse to rabbits and a horse. Ultrastructural morphology of the crescent-shaped bradyzoites was not compatable with Besnoitia besnoiti or B. jellisoni and it is proposed that the infection was caused by B. bennetti. PMID- 8410951 TI - Cabbage poisoning in ruminants. AB - A complaint of poor productivity was investigated in a herd of Jersey cattle and a small flock of Dorper sheep that had been fed cabbage for 5 to 6 months per year, over a number of years. During that time a number of lambs were lost as a result of swayback, some cows aborted, milk production was down, and although the farmer was an experienced inseminator, the conception rate in the Jerseys was low. After rectal examination and consulting the records, 26% of the cows were found to have been open for more than 150 d and 28% had been inseminated more than 3 times. The body condition of the cows ranged from poor to moderate, the hair coats were dull and some cows were anaemia. The young lambs had visibly enlarged thyroids. Haematological investigations revealed a typical Heinz body type haemolytic anaemic. In the cattle the anaemia appeared to be macrocytic, hypochromic and associated with thrombocytosis, while in the sheep it seemed to be normochromic and slightly microcytic. Plasma copper concentrations were severely depressed in all cases. Most thyroxine concentrations were within the normal range, although some were on the low side. Analysis of cabbage revealed a high sulphur content and marginally low copper and zinc levels. The main factor in cabbage which is responsible for poor animal performance, is the high sulphur content. High levels of this mineral exert detrimental effects, in order of priority, by elevating s-methyl cystine sulphoxide, diminishing the availability of copper and raising the glucosinolate content of the plant. PMID- 8410952 TI - Interrelationships between victimization experiences and women's alcohol use. AB - The interrelationships between experiences of childhood victimization and the development of women's alcohol-related problems are explored. Two different forms of childhood victimization are examined: (1) parent-to-child violence and (2) childhood sexual abuse (familial and nonfamilial). Data were collected from 472 women between the ages of 18 and 45 during in-depth interviews. Women were grouped to allow for the following two sets of comparisons: comparison 1: alcoholics in alcoholism treatment (n = 98), drinking drivers (n = 100) and a household sample (without alcohol problems) (n = 82); comparison 2: women with alcohol problems in treatment (n = 178), women without alcohol problems in treatment (n = 92) and a household sample (n = 82). High rates of childhood victimization for women with alcohol problems suggest that there is a link between victimization and the development, specifically, of women's alcohol problems. The strength of the interrelationships between childhood victimization and the development of women's alcohol problems when holding the treatment condition constant is of particular interest in this study. The rates of childhood victimization were significantly greater for women with alcohol problems in treatment as compared to women without alcohol problems in treatment. Thus, even when holding the treatment condition and family background variables constant, childhood victimization has a specific connection to the development of women's alcohol problems. These findings remained significant even when controlling for demographic and family background differences, including parental alcohol problems. PMID- 8410953 TI - Alcohol and violent pornography: responses to permissive and nonpermissive cues. AB - Both alcohol consumption and violent pornography have been implicated independently in the commission of sexual aggression. In addition to alcohol consumption, the presence of alcohol in the context of violent pornography may act as a permissive cue to influence judgments of such material's acceptability and self-reported likelihood of engaging in sexually violent behavior. To test this proposition, an experiment which varied beverage condition (alcohol vs no alcohol), expectancy set (expect vs do not expect alcohol) and a permissive (presence of alcohol) vs nonpermissive (absence of alcohol) cue was conducted on both male and female subjects. Secondary analyses on male subjects alone investigated the role of the personality trait hypermasculinity in affecting judgments. Results showed that (1) alcohol itself rather than expectancy set influenced judgments and willingness to engage in sexual violence; (2) the presence of alcohol interacted with alcohol consumption to affect willingness to engage in sexual violence; (3) men high in hypermasculinity judged the violent pornographic story more positively than did men low in this trait; and (4) alcohol interacted with hypermasculinity to affect acceptability judgments, but only for men low on this trait. These findings are consistent with cognitive disruption models and show promise for future investigations of the influence of permissive cues, as well as the role of hypermasculinity. PMID- 8410954 TI - A biosocial model of the alcohol-aggression relationship. AB - Four of alcohol's dose- and rate-dependent pharmacological properties may increase the likelihood of human aggression. As an anxiolytic, alcohol is capable of reducing the inhibitory effect of fear on manifestation of aggressive behavior. As a psychomotor stimulant, alcohol can potentiate aggressive behavior, once evoked, or lower the threshold for such evocation. Alcohol-related disruption of certain higher order cognitive functions may reduce the inhibitory control generally exercised by previously established knowledge and decrease ability to plan in the face of threat or punishment. Finally, alcohol's ability to increase pain sensitivity may increase the likelihood of defensive aggression. Discussion of the nature and relevance of these pharmacological properties is structured according to a heuristic and synthetic schema, predicated upon consideration of an inhibitory neuropsychological structure--the individually and culturally determined general expectancy set. PMID- 8410955 TI - Diversity of animal models of aggression: their impact on the putative alcohol/aggression link. AB - Many attempts have been made to "model" the putative link between alcohol and "aggression" using a variety of situations employing a wide range of animal species. In general, these attempts have proved somewhat disappointing. One reason for this appears to be the fact that the different "aggression" tests tap varied mixtures of offensive, defensive and predatory motivations (i.e., they are not homogenous). Indeed, one can make the claim that the majority of examples of "aggression" associated with alcohol-ingesting humans are inappropriate (nonadaptive) responses whereas many animal models tap adaptive territorial or socially competitive actions. A further problem is that alcohol has very wide ranging effects on neurophysiology and endocrine functioning. Recent evidence from studies on infra-human animals and clinical situations suggests that some of alcohol's effects on social conflict are expressed by actions in signaling and perception rather than motivation. This aspect is briefly examined in some studies with laboratory mice. PMID- 8410956 TI - Alcohol, aggression and the stress of subordination. AB - In individual resident-intruder or novel arena confrontations alcohol enhancement of aggression appears less robust than when groups of animals receive the drug. Patterns of effects in the latter situations, such as enhanced attack on familiar animals, particularly familiar females, suggest that group living engenders attack inhibitions, perhaps based on enhanced anxiety/defensiveness, that may be overcome by alcohol. Male rats living in natural, mixed-sex groups in seminatural situations develop strong dominance-subordination relationships that appear to strongly stress subordinates. Subordinate males show strongly and chronically increased defensive behaviors, increased corticosterone and reduced testosterone levels and, often, early mortality. When allowed access to both alcohol and water, they increase voluntary alcohol consumption, in comparison to dominants. Analyses of relationships between voluntary consumption and defensive behaviors support a view that the voluntary consumption increases of subordinate rats are mediated by the anxiolytic effect of alcohol in highly stressed animals. PMID- 8410957 TI - Ethanol-induced enhancement of defensive behavior in different models of murine aggression. AB - The effects of alcohol on agonistic behavior in mice were studied by introducing an intruder mouse to a resident, alcohol-treated test animal (or saline-injected control). Alcohol (0.1-2.0 g/kg, IP) was administered 20 minutes before testing, and an ethological analysis was made of all behavioral elements shown by the treated animal during a 500-second period. Alcohol did not increase aggression, whether baselines were high, low or experimentally suppressed. Defensive activities, however, were dose-dependently increased, with a threshold dose of 0.5 g/kg or lower in some situations. This suggests that alcohol did not reduce "anxiety" or "fear." Aggression tended to decrease, even with doses as low as 0.5 g/kg, which produced BACs of only 25-40 mg/dl at the start of the testing period. With the highest dose, however, aspects of timidity were still increased after 3 hours, but aggression returned to control level after 1 hour, when the BAC was about 250 mg/dl. In other studies, increased aggression has generally been found only with low alcohol doses. This acute tolerance to the anti-aggressive effect of alcohol reported here suggests the possibility of finding pro-aggressive effects at much higher BACs, perhaps more closely simulating the human situation. PMID- 8410958 TI - Brain serotonin, type II alcoholism and impulsive violence. AB - There is ample evidence that low CSF 5-HIAA concentration is associated with a tendency to exhibit impulsive violent behavior under the influence of alcohol. This is especially true for subjects who fulfill criteria for psychiatric diagnoses often associated with early onset alcoholism such as antisocial personality disorder and intermittent explosive disorder. Brain serotonin turnover as indicated by CSF 5-HIAA concentration does not correlate with CSF free testosterone concentration. The latter is more strongly associated with outward-directed aggressiveness and lack of socialization than impulsiveness. CSF ACTH concentration on the other hand correlates positively with socialization. PMID- 8410959 TI - Alcohol, benzodiazepine-GABAA receptor complex and aggression: ethological analysis of individual differences in rodents and primates. AB - Research in animals has only recently been successful in reliably mimicking the long-established link between alcohol and heightened aggressive behavior. The present review highlights the large individual differences in the effects of acute low alcohol doses on aggressive behavior in rodent and primate species, paralleling the human condition. Subpopulations of both species show reliable and repeatable enhancement of aggressive behavior when administered low, acute alcohol doses. Statistical analysis of the temporal patterns of aggressive behavior indicate that alcohol prolongs aggressive bouts or "bursts" and increases the number of aggressive behaviors within each burst. However, the latency to initiate attack and the time between aggressive bursts are relatively unaltered by alcohol. These alcohol-induced increases in aggression can be potentiated by benzodiazepine agonists and prevented by antagonists. In addition, highly aggressive animals can be differentiated from nonaggressive ones at the GABAA-benzodiazepine receptor complex. These data suggest an important link between alcohol, aggression and the GABAA-benzodiazepine receptor complex. PMID- 8410960 TI - Ethanol prevents desensitization of 5-HT2 receptor-mediated responses consequent to defeat in territorial aggression. AB - The effect of intruder status in territorial aggression on behavior in the elevated plus-maze, the open field and on 5-HT2 receptor-mediated behaviors was evaluated in male Long-Evans hooded rats. Intruders (350 g) were placed in the home cages of aggressive resident rats (475-600 g) and removed after 20 roll tumble fights. On the following day, the rats were tested on the elevated plus maze, and behavior in the open field was evaluated after injection with the 5 HT2/1C receptor agonist, DOI (1.0 mg/kg). The number of headshakes following DOI injection are thought to be an indicator of 5-HT2 receptor function. Although several other stressors were evaluated, only defeat in territorial aggression caused a significant decrement in the number of headshakes following DOI injection. To determine if ethanol (ET) could decrease the behavioral consequences of defeat, the effect of ET (1.25 g/kg) given before and immediately after aggression on behavior 24 hours later was evaluated. Although ET treatment had no effect on the control group that did not experience aggression, the ET treatment attenuated the desensitization of 5-HT2 receptor-mediated responses induced by aggression and severely exacerbated the anxiety-like effects as measured in the elevated plus-maze. These data suggest that (1) 5-HT2 receptor sensitivity decreases as a consequence of defeat, but not after several other stressors; (2) defeat in territorial aggression results in significant anxiety like effects in the elevated plus-maze 24 hours later; and (3) these anxiety-like effects are exacerbated when ET is given before, and immediately after, the aggression.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8410961 TI - What do experimental paradigms tell us about alcohol-related aggressive responding? AB - This article reviews the acute effects of alcohol on aggressive responding. The review is restricted to experimental research that use human subjects. It is concluded that a moderate dose of alcohol does not increase aggression if subjects are unprovoked. Under provocative conditions aggression is increased as a function of alcohol intoxication provided that subjects are restricted to an aggressive response. If subjects also have access to a nonaggressive response, no increase in aggression is observed. Disinhibition and arousal cannot explain the empirical results. A model assuming changes in attentional processes is a more promising explanation. PMID- 8410962 TI - The combination of alprazolam and alcohol on behavioral aggression. AB - Forty-eight moderate social drinkers were assigned to one of four treatments: alprazolam (1 mg) and alcohol (0.5 g/kg); alprazolam (1 mg) and placebo drink; placebo capsule and alcohol (0.5 g/kg); and placebo capsule and placebo drink. Breath alcohol concentrations and ratings of mood and intoxication were completed at 90, 150 and 210 minutes post-drug (45, 105 and 165 minutes post-alcohol). Subjects competed in a competitive reaction time task at 105 minutes post-drug during which psychophysiological measures were simultaneously monitored. Active treatments increased sedation and intoxication and the task increased feelings of hostility and anxiety in all subjects. Aggressive responding increased in all groups in response to provocation but some stress response dampening was shown after both alcohol and alprazolam on the psychophysiological measures and after alprazolam on subjective ratings of anxiety. The combination of alprazolam and alcohol increased behavioral aggression more than would have been predicted from the sum of the single effects, confirming clinical reports of behavioral dyscontrol. PMID- 8410963 TI - The effects of alcohol on free-operant aggressive behavior. AB - Although strong correlations between alcohol use and the frequency and intensity of social aggression have been observed in many populations, the relationship between these two events remains complex. Aggressive behavior is not always associated with alcohol or drug use, and substantial amounts of alcohol are consumed in social settings without the occurrence of violent or aggressive behavior. The relationship between alcohol and human aggressive behavior has been investigated using a free-operant procedure developed from the experimental tradition of behavioral pharmacology. The reliability of the free-operant procedure is evidenced by the similarity in experimental outcomes across laboratories using similar experimental procedures. Evidence for the validity of the procedure comes from recent studies with populations varying in histories of violent behavior, as well as from similarities in the outcome of studies using different experimental paradigms. Recent studies indicate clearly that both antecedent and consequent variables, such as the schedule of provocation and the response requirement for both aggressive and nonaggressive behavior, as well as the social context in which provocation occurs, influence the relationship between alcohol and aggressive behavior. The implications of the role of contextual factors for social policy regarding violence and alcohol use are discussed. PMID- 8410964 TI - Adolescents, alcohol and aggression. AB - Research findings demonstrate that the majority of young adults consume alcohol and that males drink more and more often than females. A significant number of high school seniors and college students have consumed five or more drinks in a row during a 2-week period. High-risk reasons for consumption include: to become intoxicated, to cope with a problem, because of anger and frustration. Social and behavioral consequences of alcohol use affect a number of young adults. Medium to heavy drinkers expect to experience more aggressiveness after drinking. Common risk factors for serious chronic delinquents and frequent users of drugs include psychological and personality factors as well as family conflict, peer factors and school failure. Although research has not determined that alcohol/drugs cause crime or produce the motivation to commit crimes, a relationship between alcohol/drug use and aggressive behavior is apparent. Alcohol plays a significant role in adolescent deaths due to accidents, homicides and suicides, acts of sexual aggression, and criminality. Implications of the research findings are that programs need to be designed with a clear philosophy and realistic goals and they need to target at-risk adolescents. Implementation of promising prevention strategies should take place in the home, school and community, incorporating the influence of parents and peers. PMID- 8410965 TI - A longitudinal investigation of alcohol use and aggression in adolescence. AB - Data from a prospective, longitudinal study of males and females tested at age 12, 15 and 18 years are used to study the relationship between alcohol use and aggression. Prevalence rates for alcohol use are similar for males and females. However, prevalence rates for aggressive behavior and alcohol-related aggression among females are lower than those for males and too low to permit meaningful analysis. Two series of nested structural equation models examine the interrelationships between alcohol use and aggressive behavior over time for all males in the sample and for male alcohol users only. The findings indicate that early aggressive behavior leads to increases in alcohol use and alcohol-related aggression, but that levels of alcohol use are not significantly related to later aggressive behavior. Thus, the data suggest that alcohol-related aggression is engaged in by aggressive people who drink. These data lend support to other research that indicates that early aggressive and antisocial behavior is predictive of later alcohol-related problems. PMID- 8410966 TI - Alcohol, drugs and human physical aggression. AB - Research on the relationship between alcohol, drugs and aggression is reviewed. The findings indicate that alcohol is a potent antecedent of aggressive behavior. Studies conducted in our laboratory demonstrate that aggressive behavior is related to the quantity of alcohol ingested, that the effect of social pressure to aggress and of intense provocation is enhanced by alcohol, that the instigating effect of alcohol depends upon the aggressive disposition of the alcohol consumer, that the aggressive behavior of the intoxicated person can be regulated by altering cues that affect cognitive processes and that other depressant increase aggressive responding. A hypothetical model is described that summarizes the experimental findings and provides a vehicle for discussing the major factors and psychological processes involved in alcohol-induced aggression. PMID- 8410967 TI - Alcohol, women and the expression of aggression. AB - The need for agreed upon definitions of terms like aggression, anger and violence is emphasized as well as the need to encompass a broader range of human aggressive behaviors in research than has been customary. Gender differences and female-as-aggressor and as object of aggression are reviewed. Some of the work on effects of alcohol on aggressive response and male and female studies are also reviewed. Some suggestions for future development of alcohol/aggression studies are made. PMID- 8410968 TI - Alcohol and anxiety: effects on offensive and defensive aggression. AB - Inconsistencies in the effects of alcohol on aggression in rodent models suggest that this effect is mediated through some other factor that is differentially involved in the various tests. The patterning of alcohol enhancement of aggression suggests that this may be most apparent in tests in which defensiveness or anxiety act to reduce aggression. Thus, an understanding of the relationship between alcohol and aggression may also involve determination of alcohol effects on anxiety. New ethoexperimental models of anxiety in rodents involve the measurement of a range of defensive behaviors to approaching, contacting predators, or to situations associated with (absent) predators. A Fear/Defense Test Battery, measuring the former, showed little, and inconsistent, response to traditional (benzodiazepine) or nontraditional (5-HT1A agonist) anxiolytics. However, an Anxiety/Defense Test Battery, measuring the latter, produced an "anxiolytic profile" of changes seen consistently to both traditional and nontraditional anxiolytics, but not to nonanxiolytic drugs. Alcohol (0.6 and 1.2 g/kg) altered the four behaviors of the "anxiolytic profile" in a manner consistent with the effects of diazepam (2.0 and 4.0 mg/kg), indicating that it is also anxiolytic. The consistency of alcohol and diazepam effects on anxiety provide a possible mechanism for their somewhat similar effects on aggression. However, alcohol at nonsedative doses, but not diazepam, additionally enhances defensive attack. Although defensive attack is behaviorally and neurally different from offensive aggression, the two are not separated in analyses of human "aggression," suggesting that alcohol effects in the latter may also be mediated by changes in defensive attack. PMID- 8410969 TI - Alcohol and premarital aggression among newlywed couples. AB - The present study examined the relationship between husband and wife alcohol use and premarital aggression, and evaluated a social learning model of alcohol and aggression that posited a moderating role for alcohol beliefs and aggressive motivations. Couples (age 18 to 29) entering their first marriage were approached after they had applied for a marriage license and interviewed concerning demographic factors and premarital aggression. These couples were recruited for the longitudinal study and given questionnaires assessing hostility, marital dissatisfaction, alcohol use and alcohol beliefs. Overall, 76% of the couples who agreed to participate returned the questionnaires. Husbands (n = 607) were approximately 24 years old, and wives were 1 year younger. Reflecting the urban setting, 72% were white and 24% were black. The results indicated a significant relationship between husband heavy alcohol use and premarital aggression. In addition, there were significant interactions between husband heavy alcohol use and marital dissatisfaction, and between husband heavy alcohol use, husband hostility and husband belief in alcohol as an excuse for aggression. These results suggested that alcohol use and premarital aggression are associated even in the absence of alcohol beliefs, but that the presence of appropriate beliefs strengthens the association. PMID- 8410970 TI - Structure-activity relationships of C-terminal endothelin hexapeptide antagonists. AB - The discovery of selective endothelin (ET) receptor antagonists will facilitate identification of the physiological and pathological roles for ET and its isopeptides. Structure-activity studies of the C-terminal hexapeptide of ET have been carried out to elucidate those amino acids important for receptor binding and agonist or antagonist activity. Binding studies were performed in rat heart ventricle, rabbit renal artery vascular smooth muscle cells, and rat cerebellum. In addition, many of the compounds have been evaluated functionally for their effects on endothelin-1-induced arachidonic acid release and inositol phosphate accumulation in specific cell lines. Selected compounds have been evaluated in a functional bioassay in tissue preparations specifically expressing either ETA or ETB receptors. We have previously described the structure-activity relationships in the hydrophobic C-terminal hexapeptide of ET, a region known to be highly important for receptor recognition. A mono-D-amino acid scan of the ET[16-21] revealed that substitution at His gave rise to analogs with significantly enhanced binding affinity. We have further evaluated the C-terminal region and will describe the design, synthesis, and pharmacological evaluation of several novel and potent ET peptide receptor antagonists. PMID- 8410971 TI - Vanilloids. 1. Analogs of capsaicin with antinociceptive and antiinflammatory activity. AB - As part of a program to establish structure-activity relationships for vanilloids, analogs of the pungent principle capsaicin, the alkyl chain portion of the parent structure (and related compounds derived from homovanillic acid) was varied. In antinociceptive and antiinflammatory assays (rat and mouse hot plate and croton oil-inflamed mouse ear), compounds with widely varying alkyl chain structures were active. Short-chain compounds were active by systemic administration in the assays mentioned above but they retained the high pungency and acute toxicity characteristic of capsaicin. In contrast, the long chain cis unsaturates, NE-19550 (vanillyloleamide) and NE-28345 (oleylhomovanillamide), were orally active, less pungent, and less acutely toxic than capsaicin. The potential of these compounds as antiinflammatory/analgesic agents is discussed in light of recent data on the mechanism of action of vanilloids on sensory nerve fibers. PMID- 8410972 TI - Conformational effects on retinoid receptor selectivity. 1. Effect of 9-double bond geometry on retinoid X receptor activity. AB - A major challenge is the development of retinoids with selective biological activities. Recently, studies on retinoid response mechanisms indicate that retinoids activate two classes of nuclear receptor proteins, the retinoic acid receptors (RARs) and the retinoid X receptors (RXRs). Here, we analyze the activity of a series of (E)- and (Z)-stilbenecarboxylic acids for gene transcriptional activation of the RARs and RXR-alpha to determine the optimum pharmacophore for receptor activation. The data obtained indicate that RAR and RXR response pathways can be separated by using the appropriate ligand. The conformations of (Z)-4-[2-(5-,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2 naphthalenyl)prop en-1-yl]benzoic acid (Z)-4-[1-(5,6,7,8-tetrahydro-5,5,8,8 tetramethyl-2- naphthalenyl)propen-2-yl]benzoic acid were examined by experimental and theoretical methods to establish the appropriate conformation of the latter that specifically activated the retinoid RXR. A palladium(0)-catalyzed aryl bromide-arylboronic acid coupling under nonanhydrous conditions was used to construct a biaryl bond in the conformationally restricted retinoid 2'- (5,6,7,8 tetrahydro-5,5,8,8-tetramethyl-2-naphthaleny)biphenyl-4-c arboxylic acid, which had RXR activity. PMID- 8410973 TI - Specificity in the binding of inhibitors to the active site of human/primate aspartic proteinases: analysis of P2-P1-P1'-P2' variation. AB - To understand the differences in the binding specificities within the aspartic proteinase family of enzymes, we have carried out studies to determine the inhibition constants of a set of related compounds with various members of the human enzyme family. The inhibition constants (Ki values) were determined by competitive inhibition of the hydrolysis of chromogenic octapeptide substrates in the pH range of 3-5. For comparison, inhibition of monkey renin was studied by RIA at pH 6.0. All inhibitors were based on the general structure 4 (morpholinylsulfonyl)-L-Phe-P2-(cyclohexyl)Ala psi[isostere]-P1'-P2'. The isosteric replacements of the scissile peptide bond included difluorohydroxyethylene, 1,2-diols, 1,3-diols, and difluoroketones. Side chain substituents in P2 include hydrogen, allyl, ethylthio, (methoxycarbonyl)methyl, N methylthiouridobutyl, imidazolylmethyl, and 4-amino-2-thiazolylmethyl. Our measurements have identified potent and selective inhibitors which are useful in evaluating the differences in the specificities among selected enzymes of this family. PMID- 8410974 TI - Synthesis and antibacterial activity of a new series of tetracyclic pyridone carboxylic acids. AB - A series of novel tetracyclic pyridone carboxylic acids replacing the 10-position oxygen atom of 9,1-(epoxymethano)-7-fluoro-8-(4-methyl-1-piperazinyl)-5-oxo-5H thiazolo [3,2-alpha]quinoline-4-carboxylic acid by imino groups (NR; R = Me, Et, c-Pr, allyl, Ph, benzyl), a sulfur atom, or a carbonyl group was prepared and evaluated for antibacterial activity and inhibitory activity on DNA gyrase isolated from E. coli KL-16. The in vitro antibacterial potency and DNA gyrase inhibitory activity were found to be in the following order: NMe > or = O > S >> C = O. Moreover, a methyl group was the optimal alkyl substituent at the 10 position nitrogen atom for antibacterial activity and for DNA gyrase inhibitory activity. 7-Fluoro-9,1-[(N-methylimino)methano]-8- (4-methyl-1-piperazinyl)-5-oxo 5H-thiazolo[3,2-alpha]quinoline-4-carboxy lic acid (10-NCH3) showed potent in vivo antibacterial activity. PMID- 8410975 TI - Structure-activity relationships of beta-D-(2S,5R)- and alpha-D-(2S,5S)-1,3 oxathiolanyl nucleosides as potential anti-HIV agents. AB - The beta-D-(2S,5R)- and alpha-D-(2S,5S)-1,3-oxathiolanylpyrimidine and -purine nucleosides with natural nucleoside configuration were synthesized and evaluated against HIV-1 in human peripheral blood mononuclear (PBM) cells. The key intermediate 14, which was utilized for the synthesis of various nucleosides, was synthesized from D-mannose or D-galactose. Condensation of the acetate 14 with thymine, uracil, cytosine, and 5-substituted uracils and cytosines gave various pyrimidine nucleosides. The acetate 14 was also condensed with 6-chloropurine and 6-chloro-2-fluoropurine which were converted to various purine nucleosides. In the case of thymine, uracil, and 5-substituted uracil derivatives, most of the compounds did not exhibit any significant anti-HIV activity except 5-fluorouracil (alpha-isomer) derivative 55. Among 5-substituted cytosine analogues, 5 bromocytosine derivative (beta-isomer) 68 was found to be the most potent anti HIV agent. In the case of purine derivatives, inosine analogue (beta-isomer) 78 was found to be the most potent anti-HIV agent in the 6-substituted purines and 2 amino-6-chloropurine derivative (beta-isomer) 90 showed the most potent activity in the 2,6-disubstituted purine series. The beta-isomers of 6-chloropurine (74), adenine (76), and N6-methyladenine (77) derivatives showed similar potencies against HIV-1, and the corresponding alpha-isomers also exhibited significant anti-HIV activity, although they were generally less potent than the beta isomers. PMID- 8410976 TI - Effect of trifluoromethyl and other substituents on activity of xanthines at adenosine receptors. AB - An aryl p-(trifluoromethyl) substituent increases the affinity of 1,3 disubstituted 8-phenylxanthines at A2a-adenosine receptors, while having little effect on affinity at A1-adenosine receptors. In contrast, an aryl p (trifluoromethyl) substituent has little effect on affinity of 3,7-disubstituted and 1,3,7-trisubstituted 8-phenylxanthines. An aryl p-sulfo substituent reduces affinity of all 8-phenylxanthines at A1- and A2a-adenosine receptors. An 8 (trifluoromethyl) substituent markedly reduces affinity of 1,3-dialkylxanthines at both A1- and A2a-adenosine receptors. In contrast, 8-(trifluoromethyl)caffeine retains affinity for A2a-adenosine receptors, but does lose affinity for A1 adenosine receptors. 8-Bromo-, 8-acryl-, and 8-pent-1-enylcaffeines are also selective for A2-adenosine receptors, while 8-cyclobutylcaffeine is nonselective. 8-[trans-2-(tert-butyloxycarbonyl)vinylcaffeine is 20-fold selective for Aza vs A1 receptors. PMID- 8410977 TI - 2-(Quinuclidin-3-yl)pyrido[4,3-b]indol-1-ones and isoquinolin-1-ones. Potent conformationally restricted 5-HT3 receptor antagonists. AB - Several series of N-(quinuclidin-3-yl)aryl and heteroaryl-fused pyridones were synthesized and evaluated for 5-HT3 receptor affinity. In the heteroaryl series, 2-(quinuclidin-3-yl)tetrahydropyrido-[4,3-b]indol-1-one (8a) and the 4,5-alkano bridged analogues (14 and 15) displayed high 5-HT3 receptor affinity with pKi values > 9. The (3S)-quinuclidinyl isomers had > 10 fold higher affinity than the (3R)-isomers. In a series of 2-quinuclidin-3-yl)isoquinolin-1-ones, derivatives substituted with small lipophilic groups (25b-e) and with 4,5-alkano-bridges (34 36) also displayed high affinity. In particular, the hexahydro-1H benz[de]isoquinolinone (S,S)-37 was the highest affinity 5-HT3 receptor ligand prepared (pKi 10.4). A number of the high affinity ligands were shown to be potent 5-HT3 receptor antagonists in vivo as determined by inhibition of the B-J reflex in the anesthetized rat. Again, (S,S)-37 was the most active agent tested (ID50 0.02 microgram/kg i.v.), and this compound was also potent in blocking cisplatin-induced emesis in both the ferret and the dog. Computer modeling studies were performed, and previously reported 5-HT3 receptor antagonist pharmacophore models were refined to include a key lipophilic binding domain. PMID- 8410978 TI - A conformational study by 1H NMR of a cyclic pentapeptide antagonist of endothelin. AB - The selective endothelin antagonist cyclo(D-Glu-L-Ala-D-allo-Ile-L-Leu-D-Trp, BE18257B) has been synthesized via solid-phase methods and its solution conformation determined by NMR spectroscopy and simulated annealing calculations based on NOE constraints. Additional information used in the structure determination included coupling constants and chemical-shift measurements as a function of temperature. The chemical shifts of two of the NH protons (D-Glu and D-Ile) exhibit low sensitivity to changes in temperature, indicating their involvement in hydrogen-bonded interactions. The main features of interest in the solution conformation include the presence of both a type-II beta-turn and an inverse gamma-turn, with central hydrogen bonds between HN of D-Glu1 and the C = O of D-allo-Ile3 and between HN of D-allo-Ile3 and the C = O of D-Glu1. The correlation of this solution conformation to the peptide's biological activity is discussed. The data are also compared with recently derived structures for BQ123, cyclo(D-Asp-L-Pro-D-Val-L-Leu-D-Trp), another highly potent endothelin antagonist. The backbone conformations of the two cyclic peptides are found to be similar. Comparisons with literature structure-activity data suggest that these peptides may mimic structural features of the C-terminal tail of the endothelins. PMID- 8410979 TI - Amesergide and structurally related nor-D-ergolines: 5HT2 receptor interactions in the rat. AB - A series of tricyclic (nor-D) partial ergolines were synthesized via a highly convergent enantiospecific strategy, ultimately arising from a racemic tricyclic ketone. Michael addition to an acrylamide, followed by reductive methylation, afforded the key intermediate. Selective deprotection and oxidation provided the tricyclic ergoline. Vascular 5HT2 receptor interactions for the partial ergolines were dramatically reduced compared to the parent ergoline, amesergide, as determined in vivo by activation of a pressor response or blockade of 5HT-induced pressor responses in pithed rats. The desisopropyl tricyclic ergolines possessed some modest pressor activity that was unlikely to be related to 5HT2 receptor activation since these compounds did not inhibit the pressor response to serotonin. In contrast, the isopropyl tricyclic ergolines exhibited no agonist activity, but inhibited the pressor response to serotonin at 1 mg/kg i.v. The ergoline amesergide inhibited the pressor response to serotonin in doses of 0.01 0.1 mg/kg i.v. The homochiral isopropyl tricyclic ergoline was more potent as a 5HT2 receptor antagonist than the epimeric (unnatural stereochemistry) analogue. Thus, the isopropyl moiety on the indole nitrogen is important for vascular 5HT2 receptor affinity in the rat. Most importantly, these data suggest that conformational rigidity of the ergoline D-ring is required for optimal 5HT2 receptor interactions in the rat. PMID- 8410980 TI - 2-(Alkylamino)nicotinic acid and analogs. Potent angiotensin II antagonists. AB - A series of pyridines and other six-membered ring heterocycles connected to a biphenyltetrazole with a -CH2-NR'-link (1) were discovered to be potent angiotensin II antagonists. In the pyrimidine carboxylic acid series (W = CR, X = N, Y = CH, Z = COOH), compounds with an alkyl group (R') on the exocyclic nitrogen were much more potent than compounds with an alkyl group (R) on the heterocyclic ring. The corresponding pyridine, pyridazine, pyrazine, and 1,2,4 triazine carboxylic acids also showed potent in vitro angiotensin II antagonism. The pyridine (W, X, Y = CH, Z = COOH, R' = n-C3H7) demonstrated potent in vitro activity (pA2 = 10.10, rabbit aorta, and Ki = 0.61 nM, receptor binding in rat liver) as well as exceptional oral antihypertensive activity and bioavailability. Any nonacidic replacement for the carboxylic acid was detrimental for activity. PMID- 8410981 TI - Plant antitumor agents. 30. Synthesis and structure activity of novel camptothecin analogs. AB - A large number of camptothecin (CPT) analogs have been prepared in the 20S, 20RS, and 20R configurations with a number of ring A substituents. Topoisomerase I (T I) inhibition data (IC50) have been obtained by standard procedures. In general, substitution at the 9 or 10 positions with amino, halogeno, or hydroxyl groups in compounds with 20S configuration results in compounds with enhanced T-I inhibition. Compounds in the 20RS configuration were less active in vitro and in vivo and those in the 20R configuration were inactive. Compounds with 10,11 methylenedioxy substitution on ring A displayed a marked increase in potency in the T-I inhibition assay. The activities of some of the analogs as determined in a variety of in vivo assays including the L-1210 mouse leukemia assay were, in general, in accord with T-I inhibition. A number of water-soluble analogs such as 20-glycinate esters, 9-glycinamides, or hydrolyzed lactone salts were prepared and tested in in vitro and in vivo assays. In general, these compounds were less active than CPT both in terms of T-I inhibition and life prolongation in the L 1210 assay. However, certain 20-glycinate esters showed good in vivo activity after iv administration. PMID- 8410982 TI - Keto/enol epoxy steroids: a new structural class of HIV-1 Tat inhibitors. PMID- 8410983 TI - (E)-3-[[[[6-(2-carboxyethenyl)-5-[[8-(4- methoxyphenyl)octyl]oxy]-2-pyridinyl] methyl]thio]methyl]benzoic acid: a novel high-affinity leukotriene B4 receptor antagonist. PMID- 8410984 TI - Synthesis and metabotropic receptor activity of the novel rigidified glutamate analogues (+)- and (-)-trans-azetidine-2,4-dicarboxylic acid and their N-methyl derivatives. PMID- 8410985 TI - Synthesis of antimicrobial agents. 5. In vivo metabolism of 7-(4-hydroxypiperazin 1-yl)quinolones. AB - A series of novel pyridone carboxylic acids having a 4-hydroxypiperazin-1-yl, a 4 hydroxy-3-methylpiperazin-1-yl, and a 4-hydroxy-3,5-dimethylpiperazin-1-yl group was prepared, and their metabolism to corresponding piperazinyl derivatives after oral administration to mice and rats was studied. This reductive metabolism appeared to be more extensive in mice than in rats. Moreover, the introduction of a methyl group into the alpha-position of the 4-hydroxy group depressed the metabolism in both species. PMID- 8410986 TI - Preparation of substituted N-phenyl-4-aryl-2-pyrimidinamines as mediator release inhibitors. AB - The role of immunologically released mediators, such as histamine, leukotrienes, and platelet-activating factor, is well-established for asthma and other allergic disorders. Developing therapeutic agents which would block mediator release from mast cells and other relevant cell types would provide a rational approach to asthma therapy. Using human basophil as a screen, a series of 4-aryl-2 (phenylamino)pyrimidines was found which inhibited mediator release. These compounds were prepared by condensing acetyl heterocycles with dimethylformamide dimethyl acetal to form enaminones which are cyclized with aryl guanidines to give pyrimidines. After examining a large number of analogs, N-[3-(1H-imidazol-1 yl)phenyl]-4-(2-pyridinyl)-2- pyrimidinamine (1-27) was chosen for toxicological evaluation. PMID- 8410987 TI - Synthesis and antiviral activity of 2'-substituted 9-[2 (phosphonomethoxy)ethyl]guanine analogues. AB - A series of 2'-substituted derivatives of 9-[2-(phosphonomethoxy)ethyl]guanine (PMEG, 1) have been synthesized and evaluated in vitro for anti-human immunodeficiency virus (HIV) activity in the XTT assay and for anti-herpes activity in the plaque reduction assay. It has been observed that the anti-HIV activity of these derivatives depends on the size and the nature of the substituent as well as the chirality at the 2'-position of PMEG. In addition, these compounds generally demonstrated greater activity against HIV than herpes viruses. The most interesting analogues which emerged from these studies are (R) 2'-(azidomethyl)-PMEG [(R)-5] and (R)-2'-vinyl-PMEG [(R)-11]. The former showed anti-HIV activity with an IC50 of 5 microM and a cytotoxicity (CC50) greater than 1.4 mM in CEM cells. The latter has an IC50 of 13 microM for anti-HIV activity and a CC50 of greater than 1.6 mM. Furthermore, we have demonstrated that replacement of the guanine base of these 2'-substituted PMEG analogues with cytosine drastically reduces anti-HIV and anti-herpes activity. PMID- 8410988 TI - Novel antagonists of 5-HT3 receptors. Synthesis and biological evaluation of piperazinylquinoxaline derivatives. AB - A series of piperazinylquinoxalines has been synthesized and studied as 5-HT3 receptor antagonists in different preparations. Antagonism to 5-HT in the longitudinal muscle of the guinea pig ileum was particularly prominent in cyanoquinoxaline derivatives with an alkyl substitutuent on the piperazine moiety. The pA2 of some selected compounds against the 5-HT3 agonist 2-methyl-5HT in the guinea pig ileum was in the range of tropisetron or ondansetron, and one of them, 7e, was more potent than these reference compounds by approximately 2 or 3 orders of magnitude. However, these compounds were markedly less potent than either tropisetron or ondansetron as displacers of 3H-BRL 43694 binding to rat cortical membranes or as antagonists of the Bezold-Jarisch reflex in rats. Piperazinylcyanoquinoxalines represent a new class of 5-HT3 antagonists with a selective effect on guinea pig peripheral receptors. PMID- 8410989 TI - Structure-affinity relationship studies on 5-HT1A receptor ligands. 1. Heterobicyclic phenylpiperazines with N4-alkyl substituents. AB - Structure-affinity relationship (SAR) studies for 5-HT1A receptor site are presented for two series of heterobicyclic phenylpiperazines with N4-alkyl substituents: 4-alkyl derivatives of 1-(2,3-dihydro-1,4-benzodioxin-5 yl)piperazine (3) and 1-(benzo[b]furan-7-yl)piperazine (4). The linear and branched hydrocarbon chain derivatives up to n-decyl were synthesized and evaluated for their ability to displace [3H]-2-(di-n-propylamino)-8 hydroxytetralin from its specific binding sites in rat frontal cortex homogenates. All compounds displayed a nanomolar affinity for the 5-HT1A receptor. In both series the N-ethyl and N-n-propyl substituted derivatives have similar affinities, being slightly but statistically significantly less active than the N-methyl-substituted derivatives. Elongation of the hydrocarbon chain increases the affinity for the central 5-HT1A receptor site, reaching a local maximum for the N-n-hexyl-substituted phenylpiperazines 23 (Ki = 0.50 nM) and 39 (Ki = 0.54 nM). Assuming that the arylpiperazine derivatives at the 5-HT1A binding site are in the ionic state, ionization constants were determined in order to evaluate the use of the local inhibition constant, Ki+, as a more convenient parameter to study the structure-affinity relationships. However, the Ki+ could not account for the specific N4-substituent effects found. PMID- 8410990 TI - Stereospecific and selective 5-HT2 antagonism in a series of 5-substituted trans 1-piperazino-3-phenylindans. AB - A study of the effect of aromatic substitution on 5-HT2, D2, and alpha 1 receptor affinity in a subseries of new and previously synthesized 1-piperazino-3 phenylindans indicated that high 5-HT2 selectivity could be obtained in 5 substituted derivatives. Accordingly, a series of 5-substituted derivatives was synthesized with the goal of obtaining stereospecific and selective, centrally acting 5-HT2 antagonists. This goal was fulfilled in 5-chloro- or 5-fluoro substituted compounds with 2-(3-alkyl-2-oxoimidazolidin-1-yl)ethyl- or 2 (tetrahydro-2-oxo-1H-pyrimidin-1-yl)ethylpiperazine substituents, as well as in their imidazolidine-2-thione or pyrimidine-2-thione analogues. The most interesting derivatives were resolved either directly via diastereomeric salts or by syntheses from resolved starting materials. Optical purity was determined by a 1H NMR method, using the chiral shift reagent (R)-(-)-2,2,2-trifluoro-1-(9 anthryl)ethanol. The compound (-)-trans-1-[2-[4-[5-chloro-3-(4-fluorophenyl)-2,3 dihydro-1H-inden++ +-1- yl]piperazin-1-yl]ethyl]-3-isopropyl-2-imidazolidinone (( )-20) had the overall best profile with a high stereoselectivity (eudismic ratio: 68) and a high selectivity versus D2 and alpha 1 receptors (affinity ratios 182 and 191, respectively). It had a potent central effect but was shorter-acting than the tetrahydropyrimidinone or thione derivatives ((-)-39, (+)-40, (-)-41, and (+)-42). The observed activities of the compounds are settled in perspective in relation to a recently proposed D2 receptor interaction model. While there are no indications so far that trans-1-piperazino-3-phenylindans interact with D2 and 5-HT2 receptors in different conformations, the present study shows important differences in aromatic substitution effects. Only 5-HT2 receptors are able to accommodate a 5-substituent in the indan benzene ring, thus allowing syntheses of highly selective compounds. PMID- 8410991 TI - Substituted thiopyrano[2,3,4-c,d]indoles as potent, selective, and orally active inhibitors of 5-lipoxygenase. Synthesis and biological evaluation of L-691,816. AB - Thiopyrano[2,3,4-c,d]indoles are a new class of 5-lipoxygenase (5-LO) inhibitors. SAR studies have demonstrated that the thiopyran ring, the 5-phenylpyridine substituent, and an acidic functional group on a four-carbon C-2 side chain are all required for optimal inhibitor potency. In contrast, the indolic nitrogen may be substituted with a variety of lipophilic groups. As a result of the SAR investigation, 44 (L-691,816; 5-[3-[1-(4-chlorobenzyl)-4-methyl-6-[(5 phenylpyridin-2-yl)methoxy ]- 4,5-dihydro-1H-thiopyrano[2,3,4-c,d]indol-2-yl]-2,2 dimethylpro pyl]-1H- tetrazole) has been identified as a potent inhibitor of the 5-LO reaction both in vitro and in a range of in vivo models. Compound 44 inhibits 5-HPETE production by both rat and human 5-LO and LTB4 synthesis in human PMN leukocytes (IC50s 16, 75, and 10 nM, respectively). The mechanism of inhibition of 5-LO activity by compound 44 appears to involve the formation of a reversible deadend complex with the enzyme and does not involve reduction of the nonheme iron of 5-LO. Compound 44 is highly selective for 5-LO when compared to the inhibition of human FLAP, porcine 12-LO, and also ram seminal vesicle cyclooxygenase. In addition, 44 is orally active in a rat pleurisy model (inhibition of LTB4, ED50 = 1.9 mg/kg; 8 h pretreatment) as well as in the hyperreactive rat model of antigen-induced dyspnea (ED50 = 0.1 mg/kg; 2-h pretreatment). Excellent functional activity was also observed in both the conscious allergic monkey and sheep models of asthma. In the latter case, the functional activity observed correlated with the inhibition of urinary LTE4 excretion. PMID- 8410992 TI - Renin inhibitors containing a pyridyl amino diol derived C-terminus. AB - Based on the concept of transition-state analogs, a series of nonpeptide renin inhibitors with the new (2S,3R,4S)-2-amino-1-cyclohexyl-3,4-dihydroxy-6-(2 pyridyl)hexane moiety at the C-terminal functionality were synthesized and evaluated for inhibition of renin both in vitro and in vivo. All compounds exhibited potencies in the nanomolar or even subnanomolar range when tested versus human renin in vitro. Selected inhibitors were evaluated in anesthetized, sodium-depleted rhesus monkeys and produced a marked reduction in mean arterial blood pressure (MAP) upon intraduodenal administration of a dose of 2 mg/kg. Compound 38 (S 2864) containing an amino piperidyl succinic acid derived N terminal is the most promising member in this series. 38 inhibited human renin with an IC50 of 0.38 nM, did not affect other human aspartic proteinases, and decreased mean arterial blood pressure significantly by 27% with a duration of action of 90 min after administration of 2 mg/kg id in anesthetized, sodium depleted rhesus monkeys. PMID- 8410993 TI - Mammalian topoisomerase II inhibitory activity of 1-cyclopropyl-6,8- difluoro-1,4 dihydro-7-(2,6-dimethyl-4-pyridinyl)-4-oxo-3-quinolinecarb oxylic acid and related derivatives. AB - 1-Cyclopropyl-6,8-difluoro-1,4-dihydro-7-(2,6-dimethyl-4-pyridinyl)-4-ox o-3 quinolinecarboxylic acid (1), a previously reported potent inhibitor of bacterial DNA gyrase, was found to be interactive with mammalian topoisomerase II (topo II). In a DNA-cleavage assay using topo II isolated from HeLa cells, 1 exhibited an EC50 value of 7.6 microM (VP-16; EC50 = 0.81 microM). A series of analogues modified at the 1-, 2-, 3-, 5-, and 7-positions of 1 were subsequently made and assessed for topo II inhibition. Compound 1 was considerably more potent than derivatives where the 1-substituent was alkyl, aryl, or H, or when N-c-C3H5 was replaced with S. The descarboxyl (i.e., 3-H) analogue had potency comparable to that of 1; when both these compounds were substituted at the 2-position with methyl or phenyl, an interesting relationship between activity and the conformation of the carboxyl group emerged. Upon replacement of the 5-H of 1 with NH2 or F, sustained potency was seen. No enhancement of activity was evident upon replacing the 7-substituent of 1 with other pyridinyl groups, 4-methyl-1 piperazinyl, or pyrrolidinyl groups; however, the 7-(4-hydroxyphenyl) analogue (CP-115,953) was 6-fold more potent than 1. The topo II inhibitory properties of 1 translated to modest in vitro cytotoxicity and in vivo activity versus P388. PMID- 8410994 TI - Synthesis and structure-activity relationships of naphthalene-substituted derivatives of the allylamine antimycotic terbinafine. AB - Derivatives of the allylamine antimycotic terbinafine (1) with varied substitution at the naphthalene ring system have been prepared, and their antifungal activity has been evaluated. In general, the potency is strongly dependent on the bulkiness of the substituent. Only hydrogen or in some cases fluorine are tolerated as substituents at positions 2-4 and 6-8 of the naphthalene moiety, whereas 5-substituents may be larger in size (F, Cl, Br, Me). Derivatives with fluorine at positions 3, 5, and 7 or chlorine at position 5 showed enhanced activity against yeasts relative to 1. This increase in sensitivity could be intensified by simultaneous introduction of two fluoro substituents at positions 5 and 7. Compound 7q demonstrated 8- to 16-fold improved potency against Aspergillus fumigatus, Candida albicans, and Candida parapsilosis. PMID- 8410995 TI - Synthesis and evaluation of a series of benzylaniline hydrochlorides as potential cytotoxic and antimitotic agents acting by inhibition of tubulin polymerization. AB - Although certain substituted cis-stilbenes have displayed potent tubulin polymerization inhibitory activity and significant cytotoxicities in cancer cell cultures, these compounds have limited aqueous solubility and are therefore difficult to formulate for in vivo evaluation. A series of water-soluble N-(3,4,5 trimethoxybenzyl)aniline salts has therefore been synthesized in which the olefinic bridge of the stilbenes is replaced by an aminomethylene hydrochloride moiety. A relationship was found between the size of the substituent in the 4 position of the aniline ring and both antitubulin activity and cytotoxicity, such that the smaller the substituent, the greater the potency. The most promising of the newly synthesized compounds was 4-methyl-N-(3,4,5-trimethoxybenzyl)aniline hydrochloride, with an IC50 value of 3.5 microM for inhibition of tubulin polymerization and cytotoxicity for a wide variety of cancer cell lines. The cytotoxicities of the benzylaniline hydrochlorides correlated remarkably well with their antitubulin activities. PMID- 8410996 TI - Iron (III)-chelating resins. 3. Synthesis, iron (III)-chelating properties, and in vitro antibacterial activity of compounds containing 3-hydroxy-2-methyl-4(1H) pyridinone ligands. AB - The synthesis, iron (III)-chelating properties, and antibacterial activity of several compounds containing the 3-hydroxy-2-methyl-4(1H)-pyridinone (HMP) moiety are described. Using the HMP derivatives iron (III) could be mobilized from iron (III)-binding proteins at physiological pH with a rate order of transferrin > lactoferrin > ferritin. Addition of HMP-containing compounds to a growth medium at a concentration of 20 mM/L resulted in a complete inhibition of the growth of Escherichia coli and about 90% inhibition for Listeria inocua after 7 h of incubation at 37 degrees C. After inhibition of bacteria growth by the HMP derivatives growth started again when ferric ions were added to the medium, which implies that the antibacterial activity is due to a limitation of iron available to the organisms. PMID- 8410997 TI - Inhibition of bone resorption by H+/K(+)-ATPase inhibitors. AB - We have found that 3-(3-(ethoxycarbonyl)propionyl)-8-methoxy-4-((2- methylphenyl)amino)quinoline (1, CP-113,411), a reversible inhibitor of gastric H+/K(+)-ATPase (IC50 10-20 mM), is also a potent inhibitor of bone resorption by osteoclasts in a bone slice assay at concentrations as low as 10(-7) M, with an IC50 of 2 mM. By contrast, the structurally related H+/K(+)-ATPase inhibitor 2 (3 (ethoxycarbonyl)-8-methoxy-4-((2-methylphenyl)amino)quinoline) disclosed by Robins is slightly more potent as an inhibitor of the gastric enzyme (IC50 3-10 microM in our hands) but less efficacious than 1 as an inhibitor of osteoclasts in the bone slice assay at the lower concentrations (no effect at < or = 10(-6) M, IC50 4 mM). These findings suggest that osteoclasts contain an H+/K(+)-ATPase like enzyme which differs from the gastric one. PMID- 8410998 TI - Structure-activity relationships of trans-3,4-dimethyl-4-(3 hydroxyphenyl)piperidine antagonists for mu- and kappa-opioid receptors. AB - A series of racemic N-substituted trans-3,4-dimethyl-4-(3 hydroxyphenyl)piperidines were evaluated for opioid agonist and antagonist activity at mu and kappa receptors. Several highly potent mu and kappa antagonists were discovered; however, no compounds with high selectivity for either the mu or kappa receptor were identified. Importantly, no derivative was found to have significant opioid agonist activity. Two derivatives were resolved, and the activities of the enantiomers were investigated. Only a limited stereochemical effect on opioid receptor selectivities was observed. The structure-activity relationships described establish the existence of an important lipophilic binding site distal to the nitrogen for both mu and kappa receptors and confirm the pure opioid antagonist pharmacophore nature of the trans-3,4-dimethyl-4-(3-hydroxyphenyl)piperidine structure. PMID- 8410999 TI - 3,4-Dimethyl-4-(3-hydroxyphenyl)piperidines: opioid antagonists with potent anorectant activity. AB - A series of (3R*,4R*)-3,4-dimethyl-4-(3-hydroxyphenyl)piperidine opioid antagonists with varying substituents on the nitrogen were evaluated for their effect on food consumption in obese Zucker rats. Opioid affinity (mu, kappa, and delta for selected compounds) and opioid antagonist activity (mu and kappa) were characterized and compared to effects on food consumption. No compounds with high selectivity for either mu or kappa receptors were discovered. However, compounds in the series had exceptional potency as opioid antagonists and in reducing food consumption in the obese Zucker rat. In contrast, a few compounds with high potency as opioid antagonists had much weaker potency for inhibiting food consumption. (3R,4R)-3,4-Dimethyl-1-[(3S)-3- hydroxy-3-cyclohexyl-propyl]-4-(3 hydroxyphenyl)piperidine (11,LY255582) emerged as having the best activity profile, both in reducing food consumption and as an opioid antagonist. Compound 11 is a highly potent mu, kappa-, and delta-opioid antagonist with possible clinical utility as an appetite suppressant for weight loss. PMID- 8411000 TI - Single point D-substituted corticotropin-releasing factor analogues: effects on potency and physicochemical characteristics. AB - In an attempt to determine which conformational parameters are important for the biological activity of ovine corticotropin-releasing factor (oCRF), we have synthesized in significant amounts (50-200 mg) and characterized chemically, structurally (CD), and biologically, oCRF analogues with substitution of each amino acid by its corresponding D-isomer. Out of 37 of these analogues, three were found to be equipotent to, or twice as potent as, oCRF, 13 had potencies in the range from 10 to 60%, 17 had potencies ranging from 1 to 10%, and the four others had potencies less than 0.5%. None of the analogues antagonized oCRF induced release of ACTH in vitro at concentrations > or = 1000 oCRF. Since antagonists to CRF action can be generated by deletion of the first 8-14 residues, a series of CRF antagonists which exhibit significantly higher in vitro and in vivo biological potency than [Met18,Lys23,Glu27,29,40,Ala32,41,-Leu33,3 6,38] h/rCRF, [alpha-helical-CRF9-41], is also described. [D Phe12,Nle21,38,Arg36]h/rCRF, in particular, was found to be ca. 15 times more potent than alpha-helical-CRF9-41 in vitro. In the rat, however, this analogue was about as effective as alpha-helical-CRF9-41 in blocking CRF-induced decrease in mean arterial blood pressure and increase in heart rate. Its potency in blocking epinephrine release by CRF was not significantly different from that of alpha-helical-CRF9-41. In the adrenalectomized rat, [Lys36]alpha-helical-CRF(9 41) (1.7 mg/kg) blunted the effect of endogenous CRF over a 90-min period; by comparison, a similar dose of alpha-helical-CRF9-41 was effective for less than 1 h. PMID- 8411001 TI - Synthesis and relative potencies of new constrained CRF antagonists. AB - Two series of CRF antagonists with N alpha- and C alpha-methylated alanine and leucines were evaluated for their biological activities in vitro and in vivo in several systems. The poly-N-methylated analogue of alpha-helical-CRF9-41, [N alpha MeLeu10,15,27,37,N alpha MeAla22,32,41]-alpha-Hel-CRF9-41, was found to be considerably less potent than the parent non-N-methylated analogue. This result was expected on the basis that alpha-helicity was thought to be required for biological activity and the prediction that backbone substitutions on the nitrogen have a tendency to break alpha-helices (a hypothesis that was confirmed by circular dichroism). Next, a series of constrained analogues of the potent CRF antagonist, [DPhe12,Nle21,38]h/rCRF12-41, was synthesized that contained C alpha methylleucine and/or C alpha-methylalanine (Aib) residues at selected positions. Because C alpha-methylation is recognized to increase alpha-helicity, and because there is now strong NMR data suggesting that residues 6-36 assume a well-defined alpha-helix, it was expected that these analogues would be more potent. Although usual solid-phase peptide synthesis procedures were followed, success in coupling the C alpha-methyl amino acids was obtained only with a 1:1 mixture of BOP/HOBt. In vitro potencies of the synthesized compounds were measured in a collagenase dispersed anterior pituitary cell culture bioassay. Monosubstituted analogues were shown to be twice to one fourth as potent as the parent compound; while the pluri-substituted peptides were slightly less potent. This decrease in potency might be correlated to an unexpected lower helical content of the pluri substituted compounds (as determined by CD spectroscopy), as it was suggested that the bioactive conformation of the CRF was predominantly alpha-helical. Interestingly, one analogue, [DPhe12,Nle21,38,C alpha-MeLeu37]h/rCRF12-41, was found to be more potent and longer acting than the parent compound in two in vivo assays measuring ACTH release after intravenous administration to adrenalectomized rats and reversal of stress-induced delay in gastric emptying in the rat after intracisternal administration. The molecular basis for this increased duration of action and potency is being investigated. PMID- 8411002 TI - Cholecystokinin peptidomimetics as selective CCK-B antagonists: design, synthesis, and in vitro and in vivo biochemical properties. AB - Antagonists of cholecystokinin-B (CCK-B) receptors have been shown to alleviate CCK4-induced panic attacks in humans and to potentiate opioid effects in animals. The clinical use of these compounds is critically dependent on their ability to cross the blood-brain barrier. In order to improve this property, new, peptoid derived CCK-B antagonists, endowed with high affinity, selectivity, and increased lipophilicity have been developed. The affinity and selectivity of these compounds have been characterized in vitro and in vivo using guinea pig, rat, and mouse. Most of these compounds proved to be selective for the CCK-B receptor, the most potent analog, N-[N-[(2-adamantyloxy)carbonyl]-D-alpha- methyltryptophanyl] N-[2-(4-chlorophenyl)ethyl]glycine (26A), having a Ki value of 6.1 nM for guinea pig cortex membranes in vitro and a good selectivity ratio (Ki CCK-A/Ki CCK-B = 174). Furthermore, the in vivo affinity of 26A for mouse brain CCK-B receptors, following intracerebroventricular injection at different concentrations, was found to be 10 nmol. Using competition experiments with the specific CCK-B ligand [3H]pBC 264, compound 26A was shown to cross the blood-brain barrier (0.2%) after intraperitoneal administration in mice. This compound is therefore an interesting pharmacological tool to further elucidate the physiopathological role of endogenous CCK. PMID- 8411003 TI - A structure-activity study of four dopamine D-1 and D-2 receptor antagonists, representing the phenylindan, -indene, and -indole structural classes of compounds. AB - Representatives of the phenylindan, -indene, and -indole classes of compounds (3 6) have been tested for affinity for the dopamine D-1 and D-2 receptors. The compounds all display high affinities for these receptors. Conformational analysis using MM2(87) and subsequent molecular least-squares superimpositions have been performed in order to determine if the affinities of the compounds can be rationalized by a recently proposed dopamine D-2 receptor-interaction model. In spite of the different geometric and conformational properties, the compounds can be well accommodated into the model in their calculated lowest energy conformations. The molecular superimpositions allow the absolute configurations of the active enantiomers of 4 and 5 to be predicted. The present structure activity study extends the receptor-interaction model by suggesting that the receptor is not very sensitive to the orientation of the p-fluorophenyl ring in 1 and 3-6 or to the exact spatial location of the associated fluoro substituent. PMID- 8411004 TI - 3-Aryl-2-(3'-substituted-1',2',4'-oxadiazol-5'-yl)tropane analogues of cocaine: affinities at the cocaine binding site at the dopamine, serotonin, and norepinephrine transporters. AB - Previous studies have shown that 3 beta-(substituted phenyl)tropan-2 beta carboxylic acid esters possess high affinity for the cocaine binding site on the dopamine transporter both in vitro and in vivo and inhibit dopamine uptake in vitro. Since 1,2,4-oxadiazoles are excellent bioisosteres of ester groups, we have prepared several 3 beta-(substituted phenyl)-2 beta-(3-substituted 1',2',4' oxadiazol-5'-yl)tropanes (5b-h) and all four stereoisomers of (1R,5S)-3 phenyl-2 (3-methyl-1',2',4'-oxadiazol-5'-yl)tropane (5a and 6-8). The 3 alpha-phenyl-2 alpha-(3'-methyl-1',2',4'-oxadiazole) isomer 7 was prepared from a stereoselective addition of phenyllithium to (1R,5S)-2-(3'-methyl-1',2',4' oxadiazol-5-yl)-8-methyl-8- azabicyclo[3.2.1]oct-2-ene (11). The binding affinities for 5a-h and 6-8 at the dopamine, serotonin, and norepinephrine transporters were obtained. In general these bioisosteres showed potencies for the dopamine transporter similar to those of their parent esters. 3 beta-(4' Chlorophenyl)-2 beta-(3'-phenyl-1',2',4'-oxadiazol-5'-yl)tropane (5d) was the most potent analogue with an IC50 of 1.62nM. However, 3 beta-(4'-chlorophenyl)-2 beta-(3'-methoxyphenyl-1',2'4'- oxadiazol-5'-yl)tropane (5e) with an IC50 of 1.81 nM was the most selective analogue for the dopamine transporter showing NE/DA and 5-HT/DA ratios of 461 and 186, respectively. The cis- and trans-3 alpha-phenyl-2 (3'-methyl-1',2',4'-oxadiazol-5'-yl)tropanes (7 and 8), which exist in a boat conformation, have IC50 values only slightly greater than that of the 3 beta,2 beta-isomer (5a) which possesses the cocaine stereochemistry. PMID- 8411005 TI - 2-Amido-8-methoxytetralins: a series of nonindolic melatonin-like agents. AB - A series of unsubstituted and methoxy-substituted 2-amidotetralins (4a-q) was prepared and evaluated for their ability to compete for 2-[125I]iodomelatonin binding to chicken retinal membranes and for their potency to inhibit the calcium dependent release of [3H]dopamine from rabbit retina. The lead compound, 2 acetamido-8-methoxytetralin (4j), showed a moderate affinity (Ki = 46 nM) and potency (IC50 = 1.4 nM) at the melatonin receptor. The structural requirements necessary for optimal agonistic activity at the melatonin receptor are as follows. First, the amido group, which should have a small, nonbranched alkyl group, is essential for affinity, and second, the methoxy substituent at the 8 position of the 2-amidotetralin ring is essential for optimal agonistic activity at the melatonin receptor. We concluded that this series of unsubstituted and methoxy-substituted 2-amidotetralins constitutes a class of nonindolic melatonin like agents that can be used as pharmacological tools to further characterize melatonin receptors and to elucidate the mode of action of melatonin. PMID- 8411006 TI - 3-Substituted, 3-(4-pyridinylmethyl)-1,3-dihydro-1-phenyl-2H-indol-2-ones as acetylcholine release enhancers: synthesis and SAR. AB - A series of 3-substituted, 3-(4-pyridinylmethyl)-1,3-dihydro-1-phenyl-2H-indol-2 ones was synthesized and found to enhance the stimulus-induced release of neurotransmitter acetylcholine (AcCh), and by doing so, might be useful in treating cognitive disorders where the level of this neurotransmitter may be diminished in the brain, as in Alzheimer's disease. An attempt has been made to correlate the structure of the 3-substitution with the ability of the compounds to enhance the release of AcCh from the striatum region of rat brain preparations. PMID- 8411007 TI - Chemistry, binding affinities, and behavioral properties of a new class of "antineophobic" mitochondrial DBI receptor complex (mDRC) ligands. AB - The mitochondrial DBI receptor complex (mDRC; previously called the peripheral benzodiazepine receptors) is linked to the production of neurosteroids such as pregnenolone sulfate, dehydroepiandrosterone sulfate, and others. In order to gain further information as to the function of the mDRC in the brain, we have constructed and tested both in vitro and in vivo a novel series of ligands, 2 arylindole-3-acetamides. The SAR studies detailed herein delineate some of the structural features required for high affinity binding to the mDRCs. In most cases the new ligands were prepared by use of the Fischer indole synthesis. Variations in the length and number of the alkyl groups on the amide nitrogen were probed together with the effects of halogen substituents on one or both of the aryl rings. Some ligands were also synthesized for study which represent conformationally constrained versions of the parent structure. Broad screening studies revealed these indoleacetamides to be highly selective for the mDRC, since they failed to bind with any significant affinity to other receptor systems. Some of the ligands were found to exhibit Ki values in the low nanomolar range for the mDRC as measured by the displacement of [3H]4'-chlorodiazepam. A subset of these ligands was also shown to stimulate pregnenolone formation from the mitochondria of C6-2B glioma cells with an EC50 of about 3 nM. In animal experiments ligands selected for further study were found to exhibit antineophobic effects, in spite of the fact that they exhibit no direct action on GABAA receptors. Consequently, it is postulated that these ligands owe their action to an indirect modulation of GABAA receptor function, presumably by stimulation of neurosteroid production and release from glial cells, followed by neurosteroid modulation of GABA's action on the chloride ion channel conductance of GABAA receptors. PMID- 8411008 TI - Confirmation of usefulness of a structure construction program based on three dimensional receptor structure for rational lead generation. AB - Skeletal structures very similar to those of four known inhibitors were automatically output from our computer program LEGEND, based on the three dimensional structure of the active site of the target enzyme, dihydrofolate reductase. Besides them, the program output novel promising structures that are stable intra- and intermolecularly. This result strongly supports the usefulness of this method for rational lead generation. New lead compounds can be obtained, not relying on chance or trial and error, if appropriate structural selection and modification of the output structures are made. PMID- 8411009 TI - QSAR's from similarity matrices. Technique validation and application in the comparison of different similarity evaluation methods. AB - It has recently been shown that good quantitative structure-activity relationships can be obtained through statistical analysis of molecular similarity matrices. Here we extend the technique to seven additional molecular series, previously studied using Comparative Molecular Field Analysis (CoMFA) methodology. The results are used to confirm technique applicability across a wider range of QSAR problems and to compare quantitatively the ability of various similarity indices to describe biological systems. The relative merits of this technique in comparison to CoMFA are discussed. PMID- 8411010 TI - Antiviral activity of C-alkylated purine nucleosides obtained by cross-coupling with tetraalkyltin reagents. AB - 2-, 6-, And 8-alkylated (methyl, ethyl, and vinyl) adenosine analogues were synthesized by a palladium-catalyzed cross-coupling of a tetraalkyltin with the halogenated purine nucleosides. The synthesis of the 8-substituted analogues was accomplished using a transient protection procedure. The 6-alkylated-9-beta-D ribofuranosylpurines as well as 2-ethyladenosine were cytotoxic at relatively low concentrations (0.8-10 micrograms/mL). 8-Methyladenosine was a potent and selective inhibitor of vaccinia virus, whereas 8-ethyl- and 8-vinyladenosine were specifically inhibitory to respiratory syncytial virus. 8-Vinyladenosine displayed particular activity against herpes simplex virus (type 1). PMID- 8411011 TI - Inhibitors of acyl-CoA:cholesterol acyltransferase. 5. Identification and structure-activity relationships of novel beta-ketoamides as hypocholesterolemic agents. AB - A study of structure-activity relationships of substituted beta-ketoamide ACAT inhibitors I and II was performed. The results of this study suggest that whereas the beta-keto group was tolerated with no loss in activity, beta-hydroxy and oxime moieties led to significantly reduced activity in vitro and in vivo. The most potent inhibitor from the acyclic series (I) (11, IC50 = 0.006 microM) contained a C-13 alkyl chain. This compound reduced plasma total cholesterol by 38% and 66% at 3 and 30 mg/kg, respectively, in cholesterol-fed rats. Dimethylation alpha to the anilide core (5) and subsequent N-methylation of the amide NH (6) decreased in vitro potency significantly. It was also found that high potency was retained with inhibitors which incorporated the carbonyl into a lactam ring (II). PMID- 8411012 TI - Synthesis and protein-tyrosine kinase inhibitory activity of polyhydroxylated stilbene analogues of piceatannol. AB - A series of hydroxylated trans-stilbenes related to the antileukemic natural product trans-3,3',4,5'-tetrahydroxystilbene (piceatannol) (1) has been prepared and tested for inhibition of the lymphoid cell lineage-specific protein-tyrosine kinase p56lck, which plays an important role in lymphocyte proliferation and immune function. A number of the analogues displayed enhanced enzyme inhibitory activity relative to the natural product. Reduction of the double bond bridging the two aromatic rings and benzylation of the phenolic hydroxyl groups was found to decrease activity significantly. The most potent compounds in the series proved to be trans-3,3',5,5'-tetrahydroxystilbene, trans-3,3',5 trihydroxystilbene, and trans-3,4,4'-trihydroxystilbene. PMID- 8411013 TI - Novel hypotensive agents from Verbesina caracasana. 2. Synthesis and pharmacology of caracasanamide. AB - Caracasanamide, one of the hypotensive agents isolated from Verbesina caracasana, is a mixture of (Z)-1a and (E)-1b forms of 1-[(3,4-dimethoxycinnamoyl)amino]-4- [(3-methyl-2-butenyl)-guanidino]butane. The structure of (E)-caracasanamide (1b) was confirmed by high-yielding synthesis starting from N,N'-bis(tert butoxycarbonyl)-S-methylisothiourea. The water-soluble Z-form of 1a, assayed by i.v. route in anesthetized rats at doses ranging from 50 to 1600 micrograms/kg body weight, was found to decrease blood pressure, to increase cardiac inotropism, respiratory frequency, and tidal volume, and to induce a very slight and not significant tachycardia. Higher doses determined respiratory depression and, in some cases, consequent cardiac arrest. The compound was shown to affect cardiovascular function by acting at the vascular level in inducing arterial vasodilation, by determining sympathetic hypotone through central neurogenic mechanisms, and by interacting with the cardiac beta 1-adrenoreceptors. The respiratory effects were independent of the cardiovascular ones. In lowering blood pressure, the compound was more potent than guanethidine and not less potent than reserpine and papaverine. (Z)-Caracasanamide may therefore be useful in the treatment of arterial hypertension of moderate degree. PMID- 8411014 TI - Imidazol-1-yl and pyridin-3-yl derivatives of 4-phenyl-1,4-dihydropyridines combining Ca2+ antagonism and thromboxane A2 synthase inhibition. AB - A series of derivatives of 4-phenyl-1,4-dihydropyridine bearing imidazol-1-yl or pyridin-3-yl moieties on the phenyl ring were synthesized with the aim of combining Ca2+ antagonism and thromboxane A2 (TxA2) synthase inhibition in the same molecule. Some of these compounds showed significant combined Ca2+ antagonism and TxA2 synthase inhibition in vitro, while others showed only one single activity. Structural requirements for significant single or combined activities are discussed. Theoretical conformational analysis, by molecular mechanics and semiempirical AM1 calculations, was performed for 1,4-dihydro-2,6 dimethyl-4-[3-(1H-imidazol-1-yl)phenyl]- 3,5-pyridinedicarboxylic acid, diethyl ester (FCE 24265) and two close congeners. FCE 24265, which inhibited TxB2 production in rat whole blood with IC50 = 1.7 x 10(-7) M and antagonized K+ induced contraction in guinea pig aorta with IC50 = 6.0 x 10(-8) M, was selected for further pharmacological evaluation. Our results show that this compound is less potent than nifedipine both in vitro and in vivo yet presents a favorable profile in vivo, lowering blood pressure without inducing reflex tachycardia. Moreover, its additional potent and selective TxA2 synthase inhibitory activity makes this compound an interesting pharmacologic tool in pathologies where both enhanced TxA2 synthesis and cellular Ca2+ overload are involved. PMID- 8411015 TI - Steroidal affinity labels of the estrogen receptor. 1. 17 alpha (Bromoacetoxy)alkyl/alkynylestradiols. AB - To develop steroidal affinity labels for the estrogen receptor, we prepared five electrophilic estradiol derivatives bearing the 17 alpha-propyl, 17 alpha-(1' butynyl), or 17 alpha-(1'octynyl) chain, with either a terminal epoxy function (for the 17 alpha-propyl substituent) or a terminal bromoacetoxy function (for all three 17 alpha-substituent types). These compounds displayed low affinity for the lamb uterine estrogen receptor; with apparent relative affinity constants ranging from 0.02% to 0.24% that of estradiol. They were also rapidly transformed in cytosol, probably to the corresponding vicinal diols (epoxy compounds) or primary alcohols (bromoacetoxy compounds). Nevertheless, bromoacetates induced irreversible inactivation of the hormone-binding site but only with ligand-free binding sites. The effect of bromoacetates was prevented by treatment of the cytosol with the thiol-specific reagent methyl methanethiosulfonate. Inactivation of the receptor at 0 degrees C was rapid (< 1 h) and strongly dependent on both compound concentration and pH, with significant effects obtained at either > 150 nM (at pH 9) or pH > 7.5 (at 5 microM). Regardless of the conditions used, the order of efficiency for bromoacetates was always: 17 alpha-propyl derivative < 17 alpha-butynyl derivative < 17 alpha-octynyl derivative, with maximal inactivation of approximately 30% and approximately 70% of the hormone-binding sites obtained for the less active and more active compounds, respectively. Characteristics of the receptor inactivation suggest that (i) prepared bromoacetates are highly reactive affinity labels for the estrogen receptor, (ii) they react with similar (or even a single) nucleophilic amino acid residues located within or near the hormone-binding site of the receptor; these residues are probably the -SH of cysteines, and (iii) position 17 alpha of steroidal ligands is suitable for introducing electrophilic substituents to develop efficient affinity labels for the receptor. PMID- 8411016 TI - Synthesis and structure-activity relationships of 1-acyl-4-((2-methyl-3 pyridyl)cyanomethyl)piperazines as PAF antagonists. AB - A second generation of (cyanomethyl)piperazines, 1-acyl-4-((2-methyl-3 pyridyl)cyanomethyl)-piperazines, with increased oral activity was prepared and evaluated in vitro in a PAF-induced platelet aggregation assay (PAG) and in vivo in a PAF-induced hypotension test in normotensive rats (HYP). Oral activity was ascertained through a PAF-induced mortality test in mice (MOR). Attachment of a methyl group at position 2 of our earlier pyridine derivatives resulted in an improvement of 1 order of magnitude or greater in the ID50 of the oral test. Three different types of acyl substituents of similar potency emerge from this work: N-(diphenylmethylamino)acetyl, 3-substituted 3-hydroxy-3-phenylpropionyl, and N-substituted 3-amino-3-phenylpropionyl groups. The most interesting compounds, 26 (UR-12460, PAG IC50 = 0.040 microM, HYP, ID50 = 0.021 mg/kg i.v., MOR, ID50 = 0.30 mg/kg po) and 58 (UR-12519, PAG IC50 = 0.041 microM, HYP, ID50 = 0.015 mg/kg i.v., MOR, ID50 = 0.044 mg/kg po), compare favorably with WEB-2086. Compounds 26 and 58 were also tested in active anaphylactic shock (AAS) and endotoxin-induced mortality (EIM) tests. On the basis of these data, compounds 26 and 58 have been selected for further pharmacological development. PMID- 8411017 TI - Synthesis and evaluation of unsymmetrically substituted polyamine analogues as modulators of human spermidine/spermine-N1-acetyltransferase (SSAT) and as potential antitumor agents. AB - Spermidine/spermine-N1-acetyltransferase (SSAT), the rate-limiting step in polyamine catabolism, is critical for the interconversion and modulation of cellular polyamines. Inhibitor-initiated induction of this enzyme also appears to correlate with the sensitivity of tumor cells to a class of novel polyamine analogues, the bis(ethyl)polyamines. Thus, terminally alkylated polyamines which modulate the cellular level of SSAT could be of great value for understanding the role of this enzyme both in analogue-mediated cytotoxicity and in overall cellular polyamine metabolism. Such analogues could also become important therapeutic agents by disrupting cellular polyamine metabolism. The structure activity relationships defining the interaction of polyamine analogues with SSAT have not been fully elucidated, and, in particular, unsymmetrically alkylated polyamines have not been synthesized and evaluated as modulators of SSAT. To this end, we now report the synthesis and preliminary biological evaluation of N1 ethyl-N11-propargyl-4,8-diazaundecane and N1-ethyl-N11-((cyclopropyl)methyl)-4,8 diazaundecane via a synthetic pathway which represents an efficient route to a variety of unsymmetrically substituted polyamine analogues. The title compounds act as effective inhibitors of isolated human SSAT and produce a differential superinduction of SSAT in situ which appears to be associated with a cell specific cytotoxic response in two human lung cancer cell lines. In so doing, these analogues exhibit promising antitumor activity against cultured human lung cancer cells. PMID- 8411018 TI - Inhibition of herpes simplex virus type 1 ribonucleotide reductase by substituted tetrapeptide derivatives. AB - It is known that peptides corresponding to the C-terminus of the small subunit of herpes simplex virus type 1 and 2 ribonucleotide reductase can inhibit enzymatic activity by preventing the association of the enzyme's two subunits. In a quest for smaller, more potent inhibitors, we have conducted a structure activity investigation based on the pentapeptide H-Val-Val-Asn-Asp-Leu-OH. Potency increases of up to 4000 times (IC50 0.18 microM) have been achieved in an enzymatic assay by a combination of modifying the N-terminal valine to a diethylacetyl group, adding a methyl group to the beta-carbon of the adjacent valine, dialkylating the asparagine side-chain nitrogen and dimethylating the beta-carbon of the aspartic acid residue. In addition the relative contribution of various inhibitor functionalities to inhibitor potency has been investigated. PMID- 8411019 TI - Hydroxylated 2-(5'-salicyl)naphthalenes as protein-tyrosine kinase inhibitors. AB - The salicyl group figures prominently in several potent protein-tyrosine kinase (PTK) inhibitors, including the fermentation product lavendustin A (3), the salicylsulfonyl nitrostyryl 30, and our recently reported salicyl-containing stilbene 7. Taking compound 7 and the isomeric 8 as lead structures, bicyclic nuclei 9-12 were prepared as conformationally constrained mimetics in which the hydroxyphenyl rings of 7 and 8 are held coplanar with the stilbene ethylene bridge. A similar approach with styryl-based PTK inhibitors of structure 1 previously yielded analogues 2 with enhanced potency. In the present case, however, the resulting salicyl-containing bicyclics exhibited extremely poor inhibitory potency when examined against autophosphorylation of immunoprecipitated p56lck PTK preparations. The implications of these results are discussed as they relate to the potential ways in which salicyl-containing stilbenes may be oriented relative to styryl-based inhibitors of type 1 and to an emerging class of potent aryl-substituted bicyclic inhibitors exemplified by compound 31. PMID- 8411020 TI - N-(carboxymethyl)-N-[3,5-bis(decyloxy)-phenyl]glycine (Ro 23-9358): a potent inhibitor of secretory phospholipases A2 with antiinflammatory activity. PMID- 8411021 TI - Two new unsaturated fatty acid ethanolamides in brain that bind to the cannabinoid receptor. PMID- 8411022 TI - Deoxygenated inositol 1,4,5-trisphosphate analogues and their interaction with metabolic enzymes. (1R,2R,4R)-cyclohexane-1,2,4-tris(methylenesulfonate): a potent selective 5-phosphatase inhibitor. PMID- 8411023 TI - Psychosocial genetics: an emerging scientific discipline. PMID- 8411024 TI - What young people think and do when the option for cystic fibrosis carrier testing is available. AB - We report findings in phase II of a pilot study of cystic fibrosis (CF) carrier screening/testing by mutation analysis. Phase I has been reported elsewhere. Eligible participants in phase II (n = 815) were students (15 to 17 years of age) in public high schools. An educational component (exchange of information and discussion about common genetic disorders including CF) preceded, by one week or more, voluntary participation in the screening component which required a blood sample. The uptake rate for screening was 42%. Nine carriers (2pq = 0.0260) were identified, all with the delta F508 mutation; students were also tested for G551D, G542X, W1282X, and -549-mutations, but no carriers of these alleles were found. Carriers had positive views of the education and testing experiences. Persons identified as 'non-carriers' were also surveyed (n = 135, response rate 41%). As in phase I, the majority (83%) again understood that a negative DNA test had not excluded them from possible carrier status. Students who participated in the informational component but were not screened served here as controls in the follow up survey (n = 208, response rate 53%). Their views were similar to those of the screened non-carriers, and similar also to those held by students, adults, pregnant women, couples, and CF relatives in other communities. PMID- 8411025 TI - Prenatal screening for cystic fibrosis: psychological effects on carriers and their partners. AB - This study aimed to assess the psychological impact of screening for cystic fibrosis (CF) carrier status in a population of pregnant women. A cohort of 1798 women, who accepted the offer of testing before 18 weeks of pregnancy, filled in a self administered questionnaire seeking information on their perceived risk of carrier status and their emotional response, as well as a general health questionnaire (GHQ). Sixty-four women identified as CF carriers had partners who received a negative test result. This group and their partners were assessed, together with selected controls, on four further occasions: (1) on receiving the carrier's positive test result; (2) on receiving the partner's negative test result; (3) six weeks later; (4) six weeks after delivery. The instruments used were the GHQ and the Symptom Rating Test (SRT). When compared to control subjects, carriers showed a significant increase in generalised psychological disturbance which could be attributed specifically to symptoms of anxiety and depression during the period (average four days) that they awaited their partner's test result. On receiving a partner's negative test result, the carriers returned to control levels and maintained this equilibrium. Although there was no significant difference in generalised psychological disturbance between partners and their selected controls, partners did become significantly more anxious and manifested feelings of inadequacy while awaiting their own test result. Both male partners and male control subjects were more likely to become anxious if their partner was distressed. PMID- 8411026 TI - Five year study of prenatal testing for Huntington's disease: demand, attitudes, and psychological assessment. AB - Adult predictive and prenatal testing programmes for Huntington's disease (HD) in Canada have been available since 1986. However, the demand for prenatal testing and the reasons why some people choose not to have the prenatal test for this late onset disorder have not been well documented. In addition, the knowledge and attitudes of adult predictive testing candidates and their partners about prenatal testing are not well known nor are the psychological effects of prenatal testing well understood. As of September 1991, 425 subjects had entered the Canadian Collaborative Study of Predictive Testing and, of these, 47 subjects or their partners had become pregnant. Of this group, 14 (30%) couples requested prenatal testing, 24 (51%) couples did not want prenatal testing, and nine (19%) at risk subjects had already received a decreased risk through adult predictive testing and, therefore, were not eligible for the prenatal test. Of the 14 couples who initially requested prenatal testing, seven withdrew. Thus, demand for the prenatal test by eligible candidates was 7/38 or 18%, which is much lower than the 32 to 65% expected based on early survey data. The most frequently cited reason for declining prenatal testing was the hope that a cure would be found in time for their children. While the majority of adult predictive testing candidates (71%) in our study had accurate information about definitive prenatal testing, many (63%) did not have a correct understanding of exclusion prenatal testing. Although no serious adverse events such as suicide planning or admission to psychiatric hospital have occurred, a particular need for careful counselling was identified for those at risk candidates and their partners who have one prenatal test and feel compelled to use the test again in future pregnancies. Even though prenatal testing for HD is not requested as often originally expected, it still remains a desired option for some at risk persons and their partners. PMID- 8411027 TI - Perception of predictive testing for Huntington's disease by young women: preferring uncertainty to certainty? AB - Opinions on the implications of predictive testing for Huntington's disease were evaluated in a group of 169 women (aged 21-35 years) with interest in psychosocial issues, but with no special pre-existing knowledge or training in genetics. Predictive testing for Huntington's disease (HD) is considered to be a test case for predictive testing for other late onset diseases, monogenic as well as multifactorial disorders. In the hypothetical situation of having a 50% risk for developing HD, about half of the group expressed interest in a predictive test. As to the question of giving results of predictive tests to third parties, the group would be very reluctant to inform the employer or the insurer, but not their own family. Prenatal testing for late onset diseases was considered acceptable by half of the women; only one quarter of the total group would terminate a pregnancy of a child that might develop a late onset disease. The assessment of attitudes towards predictive testing was carried out within the context of a global evaluation of perceived advantages and disadvantages of genetic counselling. The attitudes towards predictive testing were systematically associated with perceiving 'having more certainty about the future' as an advantage of genetic counselling and with rejecting 'knowing everything in advance' as a disadvantage. PMID- 8411028 TI - Genetic risk: women's understanding of carrier risks in Duchenne muscular dystrophy. AB - This paper reports a study of 48 women (16 mothers and 32 daughters representing 28 families) who had lived with Duchenne muscular dystrophy (DMD) in their family. It looks at the way the women talked about their carrier risks during the course of an unstructured interview. It points to a significant difference between lay and health professionals' perspectives, in particular the thresholds they used to distinguish between high and low risk. A number of women, when quoting their risk in a mathematical form, confused their reproductive risks with their carrier risk, another indication of differential perceptions between the women and health professionals. There was evidence that several of the women did not retain their risk in a mathematical form but had translated it into a descriptive category which resolved their risk into greater certainty. PMID- 8411029 TI - Neurofibromatosis type 1 (NF1): knowledge, experience, and reproductive decisions of affected patients and families. AB - Eighty-one subjects (56 affected patients and 25 parents of isolated affected cases) from 63 families with neurofibromatosis type 1 (NF1) on the North Western Regional Genetic Family Register (NWRGFR) were interviewed. Patients were interviewed either before (n = 26) or after (n = 55) genetic counselling. In the group as a whole, knowledge of the clinical features and the genetic aspects of the condition was poor (mean score 7 within the range of 0 to 18). The following factors were significantly associated with higher knowledge: (1) genetic counselling, (2) higher social class, (3) child with NF1, (4) when NF1 had influenced reproductive decisions, (5) young age at diagnosis, and (6) member of a patient support group. The majority of the affected subjects perceived themselves to be more severely affected than by medical classification, with persons who had been diagnosed later in life, had a child with NF1, or who were concerned about the cosmetic aspects of the disease perceiving themselves to be more severely affected. Assessment of the psychosocial effects of NF1 at different stages of life showed that 63% of affected subjects experienced difficulties at school and 48% said that the condition, particularly cosmetic aspects, caused anxiety during adolescence (n = 54). These difficulties may have contributed to later problems with career attainment and confidence in relationships. Seventy-seven percent of parents stated that their child was experiencing difficulties at school relating to NF1 (n = 51). Of the subjects at risk of having a child with NF1 and who knew about NF1 before having their family (n = 32), 45% said that it had influenced their reproductive decisions. Of 29 subjects who were still considering children, 41% wished to have prenatal diagnosis in a future pregnancy, but only three subjects stated that they would terminate an affected pregnancy. PMID- 8411030 TI - Psychosocial issues raised by a familial ovarian cancer register. AB - A Familial Ovarian Cancer Register has been established which has recruited primarily through media publicity. In depth semi-structured interviews were carried out with 20 women who had volunteered in order to explore (1) knowledge about the disease, (2) motivations for contacting the Register, and (3) expectations. We found that interviewees were generally well informed about the symptoms of the disease as a result of their relatives' experiences. There was, nevertheless, a need for information which the Register was seen as potentially fulfilling, although most subjects gave altruistic reasons for volunteering. Only one interviewee said that the publicity about the Register had made her more anxious. Most said that their anxieties had not been affected either way by the Register. Subjects did not have a clear idea of what being on the Register would mean, although there was an expectation of screening for early signs of the disease. Many interviewees had models of familial disease which did not follow mendelian genetics. This has implications for the targetting of education and screening programmes. Other psychosocial issues raised by a register of this kind are discussed, many of which require continuing monitoring to ensure that the psychological costs do not outweight the benefits. PMID- 8411031 TI - 'Inside-out', back-to-front: a model for clinical population genetic screening. AB - Developments in DNA technology have resulted in a dramatic increase in the number of genes identified. With the localisation of a gene it is possible to devise procedures suitable for mass carrier screening programmes. Until recently mass carrier screening was only possible for a limited number of disorders, for example, Tay-Sachs disease and haemoglobinopathies. Counselling possible carriers was based on estimations of risk. The momentum towards mass carrier screening is likely to be increased by gene therapy. Carrier screening for cystic fibrosis alone will have dramatic implications for genetic service provision as 4 to 5% of the UK population carry the CF gene. The potential for genetic screening of multifactorial diseases, for example, cancers, should also be considered. The existing organisation of genetic services is likely to be inadequate. A new specialty of clinical population genetics is required. A model is proposed of clinical population genetic screening programmes, organised under a 'common umbrella' led by a public health physician, while screening and follow up will remain the responsibility of the appropriate clinician. PMID- 8411032 TI - Childhood onset autosomal dominant polycystic kidney disease in sibs: clinical picture and recurrence risk. German Working Group on Paediatric Nephrology (Arbeitsgemeinschaft fur Padiatrische Nephrologie. AB - In a systematic study on the clinical picture and genetics of cystic kidneys in children, in association with the German working group on paediatric nephrology (Arbeitsgemeinschaft fur Padiatrische Nephrologie), we have investigated 79 children with early manifestation of autosomal dominant polycystic kidney disease (ADPKD). They belonged to 64 families (64 index patients and 15 affected sibs). Early manifestation was defined in this study as clinical symptoms (hypertension, proteinuria, impaired renal function, palpably enlarged kidneys) occurring before the age of 15 years. In order to estimate the recurrence risk to sibs of a previously diagnosed patient with early manifesting ADPKD, we found that 15 out of a total of 65 sibs of the 64 index patients (45% of the theoretically expected 32.5 gene carriers) showed comparable early manifestation. Another 10 symptom free children were diagnosed sonographically as having ADPKD before the age of 18 years, so that the total number of affected sibs was 25/65 in the study group, representing 76% of the gene carriers. Although the gene in childhood manifesting ADPKD can be transmitted through both sexes, a statistically significant (p < 0.05) maternal predominance was observed (M:F = 23:41). In affected sibs ages of onset, initial presentation, and the development of complications appeared to be similar in the majority of families. Our data indicate a high recurrence risk to sibs for early manifestation of ADPKD which has important implications for genetic counselling and clinical care of affected families and gives clues to the underlying genetic mechanism of childhood onset ADPKD. PMID- 8411033 TI - Facioscapulohumeral muscular dystrophy: aspects of genetic counselling, acceptance of preclinical diagnosis, and fitness. AB - A questionnaire about the interest in and demand for preclinical diagnosis for facioscapulohumeral muscular dystrophy (FSH) was sent to 46 patients. Most stated that they would have liked to have known their diagnosis earlier in order to seek more efficient help, to avoid strenuous activities, to prepare themselves emotionally, or to choose an appropriate profession. Similar arguments were used to explain their interest in preclinical diagnosis for their children. Most patients also favoured prenatal diagnosis although only two stated they would abort a pregnancy in the case of an affected fetus. Genetic counselling had apparently little influence on family planning. According to this study, FSH does not seem to reduce reproductive performance in our population. PMID- 8411034 TI - An epidemiological study of isolated split hand/foot in Hungary, 1975-1984. AB - A population based and validated 10 year cohort of 94 cases with split hand/foot born in Hungary, 1975-1984 was evaluated. This type of congenital limb deficiency was relatively frequently (43%) associated with non-limb defects. Fifty-four cases with isolated split hand/foot are evaluated in this paper. A single limb was affected in 78% of cases. The upper limbs were 21 times more frequently affected in unimelic cases with a right sided predominance and male excess. Case control analysis indicated intrauterine growth retardation and lower socioeconomic status of parents of cases. Family study showed six familial cases with autosomal dominant inheritance among 152 first and 452 second degree relatives. All familial cases were males. PMID- 8411035 TI - Second polar body incorporation into a blastomere results in 46,XX/69,XXX mixoploidy. AB - A case of 46,XX/69,XXX mixoploidy is described. The patient had a normal 46,XX diploid karyotype in lymphocytes but a triploid 69,XXX cell line in most of her fibroblasts. In order to learn more about the underlying mechanism resulting in mixoploidy, we studied short tandem repeat polymorphisms (STRPs) in lymphocyte DNA of the patient's parents and in both lymphocyte and fibroblast DNA of the proband. The findings showed maternal origin of the supernumerary chromosome complement and are best explained by second polar body incorporation into a blastomere. PMID- 8411036 TI - Trisomy 10qter confirmed by in situ hybridisation. AB - We report a boy with multiple congenital anomalies compatible with trisomy for the distal region of the long arm of chromosome 10 and a male karyotype with one 18p+. In situ hybridisation with a cDNA for ornithine aminotransferase (OAT), whose locus maps to 10q26, confirmed the clinical suspicion of distal trisomy 10q. Subterminal localisation of the labelling signals on chromosome 10 and on the der(18) indicated the localisation of the OAT locus in the proximal part of 10q26. Two clusters of labelling signals were also found on the pericentromeric and proximal portion of the X chromosome short arm, thus confirming the presence in this region of two non-adjacent OAT pseudogenes. The phenotypic similarities of this patient to previously reported cases provide further support for the delineation of trisomy 10qter as a specific, clinically recognisable syndrome. PMID- 8411037 TI - Deletion of chromosome 2 (p11-p13): case report and review. AB - The case of a young man with del(2) (p11.2p13) is reported. Accounts of previous cases of deletion of the short arm of chromosome 2 are reviewed. Common features include mental retardation, proportional short stature and weight, dysmorphic facial features (a prominent nose, abnormal ears), and abnormal hands. Growth and developmental delay are present during the postnatal period. PMID- 8411038 TI - Two sibs with unbalanced translocations in the Waardenburg gene region. AB - We report two sibs with unbalanced translocations between chromosomes 2 and 11, both products of a paternal balanced reciprocal translocation involving bands 2q37.3 and 11q23.3 PMID- 8411039 TI - Mild phenotypic manifestation of a 7p15.3p21.2 deletion. AB - A 28 month old girl with dysmorphic features was found to have an interstitial deletion of the short arm of chromosome 7p15.3-7p21.2. The patient had ptosis, dacryostenosis, pectus excavatum, short hands, and her development was normal or mildly delayed. Craniosynostosis and growth retardation, which were present in two other patients with similar deletions, were not present. Because of the mild manifestations, this case expands the clinical spectrum of the 7p15-7p21 deletion phenotype. PMID- 8411040 TI - Extra G positive band on the long arm of chromosome 9. AB - Various heteromorphisms of the 9q heterochromatic area have been reported. In most instances, the extra G positive band is accompanied by an extra C band. We describe a family where the extra G band is totally euchromatic and does not include an extra C band. It is not clear whether these two types of variant chromosome 9 arose from a similar mechanism. PMID- 8411041 TI - Triple structural mosaicism of chromosome 18 in a child with MR/MCA syndrome and abnormal skin pigmentation. AB - A case of triple mosaicism involving chromosome 18 is described in a girl with abnormal skin pigmentation similar to hypomelanosis of Ito. The karyotype is 46,XX, -18, + del(18)(p11.23-->pter)/46,XX, -18, + idic(18)(p11.23)/46,XX, -18, + r(18). The patient displays some clinical features of monosomy 18p and a few signs of trisomy 18q. Our case illustrates a non-random association of chromosomal mosaicism with abnormal skin pigmentation. PMID- 8411042 TI - Presymptomatic testing for autosomal dominant spinocerebellar ataxia type 1. AB - Presymptomatic testing was done on four people from a large family in which an autosomal dominant form of spinocerebellar ataxia was segregating. Earlier genetic analysis had shown that in this family the disorder was tightly linked to an informative microsatellite polymorphism on chromosome 6p. Two subjects with prior risks of 50% of developing the disease had final risks after testing of 2%; the other two with prior risks of 25% had final risks of 1%. Chromosome 6p linked spinocerebellar ataxia may now be added to Huntington's disease as a late onset disorder in which genetic linkage may be used to carry out presymptomatic testing. PMID- 8411043 TI - Approaches to prenatal cystic fibrosis carrier screening. PMID- 8411044 TI - Cystic fibrosis and deafness. PMID- 8411045 TI - Cutis laxa and the Costello syndrome. PMID- 8411046 TI - Forensic medicine and the polymerase chain reaction technique. PMID- 8411047 TI - Association versus linkage studies in psychosis genetics. PMID- 8411048 TI - Association and linkage: complementary strategies for complex disorders. PMID- 8411049 TI - Genetic transmission of Alzheimer's disease among families in a Dutch population based study. AB - We evaluated age at onset and transmission patterns of Alzheimer's disease (AD) in families of 198 patients who had onset of symptoms before the age of 65 years and were diagnosed before the age of 70 years. Patients were ascertained in a population based study in The Netherlands. The results suggest that the risk of AD by the age of 90 in first degree relatives is 39% (95% confidence interval 27 to 51). By the age of 90, this risk is 2.8 (95% confidence interval 1.5-5.2) times greater than the corresponding risk of 14% among relatives of age and sex matched control subjects. Segregation analysis indicated that patterns of familial clustering are best explained by transmission of a major autosomal dominant gene with reduced penetrance and a multifactorial component. However, the single major locus model could be rejected in favour of the mixed model only when a cohort effect for heritability was allowed for. The frequency of the AD susceptibility allele was estimated to be 0.48% in the single major locus model and 0.31% in the mixed model. Although our study confirms that a dominant major gene is implicated in early onset AD, the results suggest that other genetic or perhaps non-genetic factors may account for the disease in a considerable number of patients. PMID- 8411050 TI - Origins of the fragile X syndrome mutation. AB - The fragile X syndrome is a common cause of mental impairment. In view of the low reproductive fitness of affected males, the high incidence of the syndrome has been suggested to be the result of a high rate of new mutations occurring exclusively in the male germline. Extensive family studies, however, have failed to identify any cases of a new mutation. Alternatively, it has been suggested that a selective advantage of unaffected heterozygotes may, in part, explain the high incidence of the syndrome. Molecular investigations have shown that the syndrome is caused by the amplification of a CGG trinucleotide repeat in the FMR 1 gene which leads to the loss of gene expression. Further to this, genetic studies have suggested that there is evidence of linkage disequilibrium between the fragile X disease locus and flanking polymorphic markers. More recently, this analysis has been extended and has led to the observation that a large number of fragile X chromosomes appear to be lineage descendants of founder mutation events. Here, we present a study of the FRAXAC1 polymorphic marker in our patient cohort. We find that its allele distribution is strikingly different on fragile X chromosomes, confirming the earlier observations and giving further support to the suggestions of a fragile X founder effect. PMID- 8411051 TI - Is skewed X inactivation responsible for symptoms in female carriers for adrenoleucodystrophy? AB - A study of X inactivation in 12 female carriers for adrenoleucodystrophy showed no evidence that skewed patterns are related to clinical manifestation. Other possible mechanisms to explain manifestation in females are considered. PMID- 8411052 TI - Sequence variations in the first exon of alpha-galactosidase A. AB - The alpha-galactosidase A gene (GALA), which is deficient in males with Anderson Fabry disease, is shown to be remarkably polymorphic in the 5' untranslated region. GALA contains seven exons. The first exon contains 60 bp of 5' untranslated sequence before the methionine initiation codon. Single strand conformation polymorphism (SSCP) screening has shown three polymorphic variants from the published sequence within the 60 base pairs. The sequence changes involved are C to T at -10, G to A at -12 (which removes an MspI site), and G to A at -30 (which removes a SacII site). The combined frequency of these is 10%. A further insertion-deletion polymorphism is detected by SSCP of a 400 bp fragment including exon 3. Both polymorphisms can be easily detected using small polyacrylamide gels and ethidium bromide staining. Nine of 20 women were informative for one of these polymorphisms and this simple SSCP analysis should be of great assistance in family studies of Anderson-Fabry disease. Such a high level of polymorphism has not been previously reported in the 5' untranslated region of a human gene and is unusual in any such short stretch of DNA. PMID- 8411053 TI - Classification of microphthalmos and coloboma. AB - A new classification of microphthalmos and coloboma is proposed to bring order to the complexity of clinical and aetiological heterogeneity of these conditions. A phenotypic classification is presented which may help the clinician to give a systematic description of the anomalies. The phenotype does not predict the aetiology but a systematic description of ocular and systemic anomalies improves syndrome identification. There are two major classes, total and partial microphthalmos, and a subclassification which follows the embryology of the anomalies. The aetiological classification consists of three classes: (1) genetic (monogenic and chromosomal), (2) prenatally acquired (teratological agents and intrauterine deformations), and (3) associations. Genetic disorders give rise to malformations; prenatally acquired anomalies are disruptions or deformations. The aetiological classification can be applied to other congenital birth defects and improves counselling of families. Recurrence risks vary considerably between the classes. PMID- 8411054 TI - Impact of genetic counselling after neonatal screening for Duchenne muscular dystrophy. AB - In a pilot neonatal screening programme for Duchenne muscular dystrophy (DMD) conducted in the Canadian province of Manitoba, a cohort of eight affected males was identified between 1 January 1986 and 31 December 1989. Demographic information, knowledge of DMD, reproductive outcome, and attitudes to prenatal diagnosis and neonatal screening for DMD were obtained through questionnaires distributed in May 1992 to the eight sets of parents of index cases, two high probability carrier aunts, and one high probability carrier sister. Personal interviews were subsequently conducted in the summer of 1992. Although there is overall consensus among the families in favour of routine neonatal screening for DMD, five of seven subsequent pregnancies reported in six women were not monitored by prenatal diagnosis and have resulted in the birth of two affected boys. In a comparable time interval, prenatal diagnosis was acceptable to carrier females whose affected male relatives were traditionally diagnosed at four or five years. We conclude that, although molecular genetic analysis now allows for precise diagnosis of DMD, highly accurate carrier testing and prenatal diagnosis, very early DMD carrier identification, and genetic counselling after the identification of DMD males in a population based neonatal screening programme may not be an effective way of decreasing the number of repeat cases of DMD within families or the overall population frequency of DMD. PMID- 8411055 TI - Campomelic dysplasia: evidence of autosomal dominant inheritance. AB - We present a mother and daughter with clinical and radiological findings consistent with the diagnosis of campomelic dysplasia. Milder tibial bowing and significant shortening of the phalangeal bones of both hands and feet may distinguish this from the classical autosomal recessive form of the disease. PMID- 8411056 TI - A three generation family with fibrodysplasia ossificans progressiva. AB - A family with five persons affected with fibrodysplasia ossificans progressiva (myositis ossificans progressiva) in three generations is described. This is the first well documented three generation family with this condition and provides further evidence for autosomal dominant inheritance. A wide range of phenotypic severity is apparent, from disabling ectopic bone formation and premature death to an asymptomatic adult with characteristic big toe malformations. PMID- 8411057 TI - The substitution of glycine 661 by arginine in type III collagen produces mutant molecules with different thermal stabilities and causes Ehlers-Danlos syndrome type IV. AB - Previous studies have shown that Ehlers-Danlos syndrome type IV (EDS IV) is caused by mutations of type III collagen (COL3A1). Here we have characterised the most amino-terminal glycine substitution so far described in a patient with EDS IV. A combination of peptide mapping and chemical cleavage analysis of cDNA localised the mutation in cyanogen bromide peptide CB5. Sequence analysis showed a G to A mutation, converting glycine 661 to arginine, which was a new dominant mutation. Analysis of type III collagen secreted by cultured fibroblasts showed an overmodified mutant protein with normal thermal stability. However, the intracellularly retained form melted 2 degrees C lower than normal. This indicated that molecules resulting from the same mutation can differ in their thermal stabilities. PMID- 8411058 TI - A new form of familial ataxia, deafness, and mental retardation. AB - Conditions causing familial ataxia, deafness, and developmental delay are considered in the context of describing brothers with a new disorder characterised by these clinical features. PMID- 8411059 TI - A new stable human dicentric chromosome, tdic(4;21)(p16;q22), in a woman with first trimester abortion. AB - A woman with first trimester abortion and a dicentric chromosome formed from a 4 and a 21 is described. The dicentric chromosome was stable and in the majority of cells the 21 centromere was active, while in a minority the chromosome 4 centromere was active. This shows that both centromeres were functional, but that only one functioned in any given cell. Suppression of the activity of one centromere might be the mechanism by which this dicentric chromosome achieved its stability. A dicentric formed from a chromosome 4 and a 21 has not apparently been previously reported. PMID- 8411060 TI - A new case of partial trisomy 19q (q13.2-->qter) owing to an unusual maternal translocation. AB - A new case of trisomy 19q13.2-->qter is described in a male child which was caused by a maternal balanced translocation (13;19)(p13;q13.2). The major clinical features detected in the patient included the following: facial dysmorphism, bilateral coloboma, narrow and hypoplastic nails, cardiac malformations (Fallot's tetralogy), genitourinary and gastrointestinal anomalies, and agenesis of the corpus callosum. A comparison with other reported cases of partial trisomy 19q is presented. A hypothesis is proposed to account for the involvement of p13 regions of different acrocentrics in some cases of familial translocations involving a chromosome 19. PMID- 8411061 TI - Split hand/split foot deformity and LADD syndrome in a family: overlap between the EEC and LADD syndromes. AB - A mother and daughter are reported with apparently dissimilar syndromes. The mother has a split hand/split foot deformity and the daughter a condition consistent with a diagnosis of LADD syndrome. Absence of clefting and deficient formation of saliva and tears are the main signs that differentiate the LADD from the EEC syndrome. However, no distinct feature is constant between these two autosomal dominant disorders that show great phenotypic variability. This report emphasises the overlap between the LADD and the EEC syndromes. PMID- 8411062 TI - Dominant carpotarsal osteochondromatosis. AB - Dominant carpotarsal osteochondromatosis is a particular disorder of the wrist and tibiotalar joints with abnormal bone proliferation and osteochondromas. Two patients, a mother and son, are described here; a similar condition has previously been described in seven affected members of a family. The upper and the lower limbs are affected in the same patient and the lesion can be bilateral. Autosomal dominant inheritance is a further criterion allowing the diagnosis of dysplasia epiphysealis hemimelica. PMID- 8411063 TI - Association between schizophrenia and homozygosity at the dopamine D3 receptor gene. PMID- 8411064 TI - Excess of homozygosity at the dopamine D3 receptor gene in schizophrenia not confirmed. PMID- 8411065 TI - Severity of chest disease in CF patients in relation to their genotypes. PMID- 8411067 TI - Integrated study of 100 patients with Xp21 linked muscular dystrophy using clinical, genetic, immunochemical, and histopathological data. Part 1. Trends across the clinical groups. AB - This multidisciplinary study was undertaken to record the variation in gene and protein expression in a large cohort of patients with well defined clinical phenotypes. The patients, whose ages ranged from 4 years to 66 years, spanned a wide range of disease severity. They represented the first 100 patients who had been examined in Newcastle, had undergone a muscle biopsy, and provided a blood sample for DNA analysis. The study had three aims: to observe any trends in the analyses across the clinical groups, to correlate gene and protein expression in individual patients, and to use the data collected to assess the relative usefulness of different techniques in the diagnosis and prognosis of patients with Duchenne and Becker dystrophy (DMD/BMD). In part 1, we describe the clinical assessment of the patients and the trends that were observed across the cohort. The patients were divided into seven groups. Group 1 had severe DMD (n = 21), group 2 had milder DMD (n = 20), group 3 were intermediate D/BMD patients (n = 9), group 4 had severe BMD (n = 5), and group 5 were more typical BMD patients (n = 31). Some patients were too young to be classified (n = 7) and a group of all the female patients were also classified separately (n = 7). The number of DMD and BMD patients was about equal, in accord with disease prevalence in the north of England, but an unusually high proportion were sporadic cases. Dystrophin labelling (performed with up to three antibodies) on both blots and sections increased gradually across the clinical groups. All histopathological indices, except the proportion of fat in biopsy sections, showed clear trends across the groups. PMID- 8411068 TI - Integrated study of 100 patients with Xp21 linked muscular dystrophy using clinical, genetic, immunochemical, and histopathological data. Part 2. Correlations within individual patients. AB - This report is the second part of a trilogy from a multidisciplinary study which was undertaken to record the relationships between clinical severity and dystrophin gene and protein expression. The aim in part 2 was to correlate the effect of gene deletions on protein expression in individual patients with well defined clinical phenotypes. Among the DMD patients, most of the deletions/duplications disrupted the open reading frame, but three patients had in frame deletions. Some of the intermediate D/BMD patients had mutations which were frameshifting while others were in frame. All of the deletions/duplications in the BMD patients maintained the open reading frame and 25/26 deletions in typical BMD group 5 started with exon 45. The deletion of single exon 44 was the most common mutation in patients from groups 1 to 3. Dystrophin was detected in sections and blots from 58% of the DMD patients with a size that was compatible with synthesis from mRNA in which the reading frame had been restored. Certain deletions were particularly associated with the occurrence of limited dystrophin synthesis in DMD patients. For example, 9/11 DMD patients missing single exons had some detectable dystrophin labelling compared with 10/24 who had deletions affecting more than one exon. All patients missing single exon 44 or 45 had some dystrophin. Deletions starting or finishing with exons 3 or 51 (8/9) cases were usually associated with dystrophin synthesis whereas those starting or finishing with exons 46 or 52 (11/11) were not. Formal IQ assessments (verbal, performance, and full scores) were available for 47 patients. Mean IQ score among the DMD patients was 83 and no clear relationship was found between gene mutations and IQ. The mutations in patients with a particularly severe deficit of verbal IQ were spread throughout the gene. PMID- 8411066 TI - Mendelian cytogenetics. Chromosome rearrangements associated with mendelian disorders. PMID- 8411069 TI - Integrated study of 100 patients with Xp21 linked muscular dystrophy using clinical, genetic, immunochemical, and histopathological data. Part 3. Differential diagnosis and prognosis. AB - This report is the third part of a trilogy from a multidisciplinary study which was undertaken to investigate gene and protein expression in a large cohort of patients with well defined and diverse clinical phenotypes. The aim of part 3 was to review which of the analytical techniques that we had used would be the most useful for differential diagnosis, and which would provide the most accurate indication of disease severity. Careful clinical appraisal is very important and every DMD patient was correctly diagnosed on this basis. In contrast, half of the sporadic BMD patients and all of the sporadic female patients had received different tentative diagnoses based on clinical assessments alone. Sequential observations of quantitative parameters (such as the time taken to run a fixed distance) were found to be useful clinical indicators for prognosis. Intellectual problems might modify the impression of physical ability in patients presenting at a young age. Histopathological assessment was accurate for DMD but differentiation between BMD and other disorders was more difficult, as was the identification of manifesting carriers. Our data on a small number of women with symptoms of muscle disease indicate that abnormal patterns of dystrophin labelling on sections may be an effective way of differentiating between female patients with a form of limb girdle dystrophy and those carrying a defective Xp21 gene. Dystrophin gene analysis detects deletions/duplications in 50 to 90% of male patients and is the most effective non-invasive technique for diagnosis. Quantitative Western blotting, however, would differentiate between all Xp21 and non-Xp21 male patients. In this study we found a clear relationship between increased dystrophin abundance (determined by densitometric analysis of blots) and clinical condition, with a correlation between dystrophin abundance and the age at loss of independent mobility among boys with DMD and intermediate D/BMD. This indicates that blotting is the most sensitive and accurate technique for diagnosis and prognosis. PMID- 8411070 TI - Epidermal mosaicism and Blaschko's lines. AB - To test the hypothesis that epidermal rather than dermal mosaicism determines Blaschko's lines in hypomelanosis of Ito (HI), we studied the distribution of chromosomal mosaicism in four patients. In two, mosaicism had not been detected in lymphocytes or dermal fibroblasts, but was clearly shown in epidermal keratinocytes; furthermore, the abnormal cell line was confirmed to the hypopigmented epidermis and the normal epidermis contained only normal cells. Negative findings in the other two patients might be because of mosaicism which was undetected either because it was submicroscopic or because it was present in melanocytes, which have not yet been studied. These preliminary results support the ideas that (1) Blaschko's lines represent single clones of epidermal cells; (2) in patients with HI and severe neurological involvement mosaicism, if detectable, is best shown in keratinocytes; and (3) the cytogenetic defect in epidermal cells may be directly responsible for the failure of pigmentation in HI. PMID- 8411071 TI - Uniparental disomy explains the occurrence of the Angelman or Prader-Willi syndrome in patients with an additional small inv dup(15) chromosome. AB - A patient with Angelman syndrome and a 46,XY/47,XY,+inv dup(15)(pter-->q11: q11- >pter) karyotype and a patient with Prader-Willi syndrome and a 46,XY/47,XY,+inv dup(15)(pter-->q12: q12-->pter) karyotype were investigated with molecular markers along chromosome 15. Paternal uniparental isodisomy was found for all informative markers in the first case which indicates that this, rather than the presence of the extra chromosome, is the cause of the Angelman syndrome phenotype. Similarly, the PWS patient showed maternal uniparental distomy with absence of PWS region material on the inv dup(15) chromosome. If (1) marker chromosomes are an occasional by product of 'rescuing' a trisomic fertilisation, or (2) if duplication of the normal homologue in a zygote which has inherited a marker in place of the normal corresponding chromosome 'rescues' an aneuploid fertilisation, or (3) if the presence or formation of a marker chromosome increases the probability of non-disjunction, then uniparental disomy might be found occasionally in other subjects with de novo marker chromosomes. PMID- 8411072 TI - Clinical and molecular studies in fragile X patients with a Prader-Willi-like phenotype. AB - A special subphenotype of the fragile X syndrome is reported which is characterised by extreme obesity with a full, round face, small, broad hands/feet, and regional skin hyperpigmentation. It resembles the Prader-Willi syndrome (PWS) and might therefore be named 'Prader-Willi-like'. Unlike the PWS, these PW-like fragile X patients lack the neonatal hypotonia with feeding problems during infancy followed by hyperphagia from toddlerhood. We describe five new fragile X patients and present a clinical update of three previously described patients with the PW-like phenotype. In one family, segregation of either the classical Martin-Bell or the PW-like phenotype was observed and in another family there was repeated transmission of the PW-like phenotype. Previously, one of the patients had been misdiagnosed as having classical PWS, based on clinical findings. Molecular studies of the FMR-1 gene showed the typical full mutations as seen in fragile X syndrome males. Molecular analysis of the 15q11-13 region, which is deleted in the majority of classical PWS patients, did not show any detectable abnormalities. In a group of 26 patients with suspected Prader-Willi syndrome but without detectable molecular abnormalities of chromosome 15, one fragile X patient was found. These clinical and molecular findings illustrate the necessity to perform DNA analysis of the FMR-1 gene in mentally retarded patients presenting with a PW phenotype but without the PWS specific cytogenetic/molecular abnormalities of chromosome 15. PMID- 8411073 TI - Mutational screening of the Wilms's tumour gene, WT1, in males with genital abnormalities. AB - Several lines of evidence suggest that the Wilms's tumour susceptibility gene, WT1, has an important role in genital as well as kidney development. WT1 is expressed in developing kidney and genital tissues. Furthermore, mutations in WT1 have been detected in patients with the Denys-Drash syndrome (DDS), which is characterised by nephropathy, genital abnormalities, and Wilms's tumour. It is possible that WT1 mutations may cause genital abnormalities in the absence of kidney dysfunction. We tested this hypothesis by screening the WT1 gene for mutation in 12 46,XY patients with various forms of genital abnormality. Using single strand conformation polymorphism (SSCP) we did not detect any WT1 mutations in these patients. However, in addition to the 12 patients, three DDS patients were also analysed using SSCP, and in all three cases heterozygous WT1 mutations were found which would be predicted to disrupt the DNA binding activity of WT1 protein. These results support the notion that DDS results from a dominant WT1 mutation. However, WT1 mutations are unlikely to be a common cause of male genital abnormalities when these are not associated with kidney abnormalities. PMID- 8411074 TI - Exclusion of candidate genes from a role in cleft lip with or without cleft palate: linkage and association studies. AB - Candidate genes and marker loci for cleft lip/palate (CL/P) were tested using linkage analyses and association studies. Eight British families with apparent autosomal dominant inheritance of non-syndromic CL/P participated in the linkage analyses while the association analyses involved 61 unrelated British white people with CL/P and 60 controls. The report of an association between RARA (17q21) and unrelated Australian persons with CL/P (p = 0.016) was not confirmed in British CL/P persons (chi 2 = 0.954, p > 0.1). There was also no evidence of linkage between RARA and the eight CL/P families (Z = -3.211, theta = 0.001). Linkage was excluded between familial CL/P and F13A1 (map position 6p24-25) with an observed maximum lod score of Z = -2.052 at theta = 0.05. No association was found between alleles at VIM (10p13) and the British CL/P subjects (chi 2 = 0.110, p > 0.5). Multipoint analysis excluded linkage between familial CL/P and the markers D1S65 and D1S58 which flank the Van der Woude syndrome locus with a maximum lod score of Z = -4.0. This suggests that the genetic defect underlying VWS is not the same as in non-syndromic CL/P. There was no evidence of linkage between CRTL1 (5q15) and the eight CL/P families (Z = -3.466, theta = 0.05). PMID- 8411075 TI - Nager acrofacial dysostosis. PMID- 8411076 TI - Hypomelanosis of Ito associated with mosaicism for trisomy 7 and apparent 'pseudomosaicism' at amniocentesis. AB - We describe a patient with hypomelanosis of Ito (HI) and typical characteristics. Cytogenetic analysis of fibroblast cultures showed mosaicism for trisomy 7. The trisomy cell line was first observed in an amniotic fluid culture and illustrates the problem of distinguishing pseudomosaicism from true mosaicism. PMID- 8411077 TI - Prenatal diagnosis of a hypermethylated full fragile X mutation in chorionic villi of a male fetus. AB - Fragile X syndrome, one of the most common human genetic diseases, is characterised by a unique genetic mechanism which involves dynamic mutation because of a heritable unstable DNA sequence and abnormal DNA methylation. Direct detection of the dynamic mutation and its methylation status at the DNA level would facilitate reliable tests for prenatal and postnatal diagnosis of the disease and for carrier detection. However, it has been suggested that DNA methylation can not be used as the basis for prenatal diagnosis as the CpG island is not always methylated in chorionic villus DNA. We report here a male fetus exhibiting both extensive somatic heterogeneity and abnormal hypermethylation of the full fragile X mutation in chorionic villus DNA as well as in fetal tissue DNA. Our results indicate that both somatic heterogeneity and hypermethylation of the full fragile X mutation are events that are clearly detectable in the 11th to 12th week of pregnancy. PMID- 8411078 TI - Congenital heart malformation in Yunis-Varon syndrome. AB - We describe a male infant with Yunis-Varon syndrome who has tetralogy of Fallot. This appears to be the first case of Yunis-Varon syndrome associated with congenital heart malformation. PMID- 8411079 TI - Two sibs with cleft palate, ankyloblepharon, alveolar synechiae, and ectodermal defects: a new recessive syndrome? AB - Hay and Wells in 1976 reported seven patients from four families who had an inherited condition of which the main features were ankyloblepharon, ectodermal defects, and cleft lip and palate. The inheritance pattern was determined to be autosomal dominant. This condition is known as AEC syndrome or Hay-Wells syndrome. We report a family with two sibs showing some of these features and congenital adhesions between the upper and lower jaws (alveolar synechiae). There seems to be a recessive pattern of inheritance as neither of the parents has any features of the syndrome. This could be described as a recessive form of Hay Wells syndrome with additional features or be named as a new syndrome. PMID- 8411080 TI - Limb/pelvis/uterus-hypoplasia/aplasia syndrome. PMID- 8411081 TI - Molecular characterisation of beta thalassaemia heterozygotes in Brazil. PMID- 8411082 TI - Autozygosity mapping, complex consanguinity, and autosomal recessive disorders. PMID- 8411083 TI - Therapeutic application of botulinum toxin. PMID- 8411084 TI - Fusobacteria: new taxonomy and related diseases. AB - Fusobacteria are anaerobic gram-negative bacilli. Since the first reports in the late nineteenth century, various names have been applied to these organisms, sometimes with the same name being applied to different species. More recently, not only have there been changes to the nomenclature, but also attempts to differentiate between species which are believed to be either pathogenic or commensal or both. Because of their asaccharolytic nature, and a general paucity of positive results in routine biochemical tests, laboratory identification of the fusobacteria has been difficult. However, the application of novel molecular biological techniques to taxonomy has established a number of new species, together with the subspeciation of Fusobacterium necrophorum and F. nucleatum, and provided new methods for identification. The involvement of fusobacteria in a wide spectrum of human infections causing tissue necrosis and septicaemia has long been recognised, and, more recently, their importance in intra-amniotic infections, premature labour and tropical ulcers has been reported. PMID- 8411085 TI - A new Brevibacterium sp. isolated from infected genital hair of patients with white piedra. AB - A new aerobic gram-positive non-sporeforming bacillus has been isolated from infected genital hair of patients with white piedra in association with Trichosporon beigelii. This species has been characterised morphologically, nutritionally, by DNA base composition, cell-wall analysis and cellular fatty acid profile on the basis of 14 isolates. The G+C content of DNA is 63.05 mol%. Cell walls possess meso-diaminopimelic acid (Type IV) and the sugars glucose, galactose, xylose and ribose; mycolic acids are not present. The species has a distinct colonial and microscopic morphology, is strongly proteolytic and produces methanethiol. These findings and the cellular fatty-acid profile are compatible with the genus Brevibacterium. A new species is proposed based on the following characters: colonial and microscopic growth and morphology; conditions for rod-to-coccus cycle; ribose utilisation; and tellurite reduction. The type strain has been named Brevibacterium mcbrellneri E2cr (ATCC 49030). The strong proteolytic properties may be the mechanism of pathogenesis. PMID- 8411086 TI - The production of porphyrins from delta-aminolaevulinic acid by Haemophilus parainfluenzae. AB - Porphyrin production by the haemin-independent Haemophilus parainfluenzae in the diagnostic porphyrin test, which determines the X-factor requirement in Haemophilus spp., was analysed quantitatively by applying modern high-performance liquid chromatographic (HPLC) methods. Ion-pair reversed-phase HPLC enabled the simultaneous separation of all porphyrin intermediates and their isomers of haem biosynthesis produced by the bacteria. The pH-dependence of porphyrin production and the respective composition of the porphyrins within the bacterial cells and in the supernate were investigated. A pH optimum of 6.9-8.0 for the production of porphyrins was found and there were marked differences in the porphyrin profiles at different pH values. PMID- 8411087 TI - Serological response of sheep to plasmid-encoded proteins of Yersinia species following natural infection with Y. enterocolitica and Y. pseudotuberculosis. AB - A prospective study of the serological response to natural infection with Yersinia enterocolitica and Y. pseudotuberculosis was performed in an experimental flock of sheep. A preliminary investigation with immunoblotting techniques showed that lambs infected with virulent Yersinia spp. produced antibodies to several yersinia outer-membrane proteins (yops) encoded by a virulence plasmid (pYV) of Y. enterocolitica or Y. pseudotuberculosis. Thereafter, an enzyme immunoassay (EIA) was developed to measure antibodies to yops. Criteria for interpreting the EIA were established by examining sera from a negative control population of lambs which had not been infected with Yersinia spp. since birth. Test samples comprised 25 pairs of pre- and post-infection sera from animals with bacteriologically proven infections with Yersinia spp. The results showed that infection of lambs with pYV-bearing strains of Y. enterocolitica or Y. pseudotuberculosis invariably evoked a significant antibody response to yops, even though all the infections were subclinical. No animal infected with so-called "environmental", pYV-negative Yersinia spp. seroconverted to yops. EIA with yops as antigen provided a sensitive and specific means to diagnose subclinical infection of lambs with virulent Yersinia spp. PMID- 8411088 TI - Comparison of antimicrobial susceptibility, beta-lactamase production, plasmid analysis and serum bactericidal activity in Edwardsiella tarda, E. ictaluri and E. hoshinae. AB - Antimicrobial susceptibility profiles of clinical and environmental isolates of Edwardsiella demonstrate that the three species are susceptible to beta-lactam antibiotics. All strains were susceptible to two quinolones tested and to gentamicin and doxycycline. E. tarda and E. hoshinae were resistant to clindamycin, whereas E. ictaluri was moderately susceptible. beta-Lactamase was produced by all strains of E. tarda, but not by E. hoshinae or E. ictaluri. A 54 kb plasmid was detected in six of 13 E. hoshinae strains. Five of the 10 E. tarda isolates studied gave an identical plasmid pattern of four plasmids ranging in size from 76-kb to 5.0-kb. One strain exhibited a 54-kb plasmid; four strains did not contain plasmid DNA. All E. ictaluri isolates contained a 5.7-kb and a 4.9-kb plasmid. E. tarda and E. ictaluri strains were resistant to human serum 20%; 12 of 13 strains of E. hoshinae were also serum resistant. Serum resistance may play an important part in the pathogenicity of these species. PMID- 8411089 TI - The production of HlyA toxin by Proteus penneri strains. AB - Twelve diverse strains of Proteus penneri of clinical origin all produced a calcium-dependent haemolysin, unlike most other Proteus spp. In most strains the haemolysin was secreted into the medium during early exponential growth and lysed not only of a variety of erythrocyte types from several animals including man, but also human neutrophils and human embryo lung fibroblasts. The haemolysin was a protein of 107 kDa, the same size as Escherichia coli HlyA, and it reacted with antiserum to E. coli HlyA. Because of its similarity in size, antigenicity and range of action to the HlyA virulence factor of E. coli, P. penneri HlyA is believed to be an important virulence factor for this organism. It was degradable by an EDTA-sensitive protease--probably the IgA protease--to inactive fragments. The interaction of P. penneri HlyA and IgA protease in vivo and the origin of HlyA, which has now been found in many diverse bacteria, are discussed. PMID- 8411090 TI - Intrathecal antibody synthesis in Lyme neuroborreliosis: use of recombinant p41 and a 14-kDa flagellin fragment in ELISA. AB - The intrathecal synthesis of IgM and IgG antibodies to Borrelia burgdorferi sonicate, to recombinant flagellin (41 kDa) and to a tryptic peptide of the flagellin (14-kDa fragment) was determined by ELISA in paired cerebrospinal fluid (CSF) and serum samples from 35 patients with Lyme neuroborreliosis (LNB) and in 10 patients with neurosyphilis. The antibody index (AI = QBb/QIg) was calculated from the ratio between CSF/serum quotients for specific antibodies (QBb) and total immunoglobulins (QIg). For the examination of IgG antibodies, the sonicate ELISA was performed with and without pre-absorption with Treponema phagedenis. Of 35 patients with LNB, 31 had intrathecal IgG response to B. burgdorferi demonstrated by sonicate ELISA (24 after absorption of cross-reactive antibodies), 29 had a response demonstrated by flagellin ELISA and 21 of 35 by 14 kDa ELISA. In patients with neurosyphilis the AI (IgG) was elevated in the sonicate ELISA in 7 of 10 samples (none of 10 after absorption of cross-reactive antibodies), in the flagellin ELISA in 5 of 10 samples and in the 14-kDa ELISA in none of 10 samples. Intrathecal synthesis of IgM antibodies to B. burgdorferi was demonstrated in patients with neuroborreliosis by sonicate ELISA in 20 of 35 samples, by flagellin ELISA in 16 of 35 samples and by 14-kDa ELISA in 9 of 35 samples. No intrathecal synthesis of B. burgdorferi-specific IgM could be detected by any assay in patients with neurosyphilis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411091 TI - Use of PCR-mediated amplification of Mycobacterium leprae DNA in different types of clinical samples for the diagnosis of leprosy. AB - DNA of Mycobacterium leprae, obtained by a highly efficient nucleic acid extraction procedure, was used for standardisation of the amplification of an M. leprae-specific repetitive sequence by use of the polymerase chain reaction (PCR). With pure DNA, M. leprae-specific amplification was obtained with as low as 100 ag (1 ag = 10(-18) g) of target DNA, a quantity equal to about one-tenth of the bacterial genome. Optimal processing of different types of clinical samples such as biopsy material, blood and lymph fluid, from multibacillary leprosy patients, was studied. Simple freezing-boiling cycles in the presence of Triton X100, with some additional sample-specific modifications such as pre treatment with NaOH to eliminate PCR inhibitors, was found to be sufficient to yield amplification of bacterial DNA in samples from paucibacillary patients. Clinical samples from 27 untreated leprosy patients, covering the various clinical forms of the disease, and with a bacterial index ranging from 5+ to 0, were collected and processed for PCR analysis. After hybridisation of the amplified material with a specific sequence, 25 of 27 patients analysed gave positive results for M. leprae in at least one of the samples. The potential of PCR for the diagnosis of leprosy is discussed. PMID- 8411092 TI - Lectin typing of methicillin-resistant Staphylococcus aureus from Singapore, England and Wales, and Denmark. AB - Methicillin-resistant Staphylococcus aureus (MRSA) isolates from a possible outbreak in Singapore (84) were examined together with all MRSA isolated in Denmark in 1986-1990 (58) and 14 distinct epidemic and 10 distinct single hospital strains from England and Wales. All 84 Singapore isolates were phage typed routinely and 52 isolates were further analysed together with the Danish isolates with an additional set of experimental phages and by lectin typing. The British strains, previously phage typed in the same way, were lectin typed. The following lectins were used: wheat-term agglutinin, soy-bean agglutinin, tomato lectin and Concanavalin-A. Routine phage typing of the Danish isolates showed that 41 isolates belonged to 19 different types; 17 isolates were non-typable (NT). Addition of experimental phage typing and lectin typing enhanced discrimination to 47 types. The 24 British strains could be divided into eight [corrected] "lectin types". Sixty-one of the isolates from Singapore were non typable by phage typing; the remaining 23 strains belonged to five types. Further examination of 52 isolates with the experimental set of phages and by lectin typing gave 14 closely related types; 48% of these isolates belonged to only two types. PMID- 8411093 TI - Virulence patterns of Vibrio cholerae non-O1 strains isolated from hospitalised patients with acute diarrhoea in Calcutta, India. AB - A collection of 28 strains of Vibrio cholerae non-O1 isolated during a 3-year period (1989-1991) from hospitalised patients with acute diarrhoea in Calcutta, India, were examined with regard to virulence-associated factors. Of the 28 isolates (each representing a case), 18 were isolated as the sole infecting agent; the remaining 10 were recovered as co-cultures from cases infected with V. cholerae O1. Of the strains isolated in this study, 82% could be serotyped, with serovars O5 (32.1%), O11 and O34 (14.3% each) predominant. Serovars O7, O14, O34, O39 and O97 were associated exclusively with sole infections. Two strains of V. cholerae non-O1 produced anti-cholera toxin IgG-absorbable cholera toxin (CT). Both CT-producing V. cholerae non-O1 strains hybridised with the DNA probe specific for the zonula occludens toxin (ZOT) but none of the remaining 26 strains hybridised with the ZOT probe. The majority of the strains were cytotoxic for CHO, HeLa and Vero cells, with end-point titres of 4-512. Fewer strains produced a cytotonic effect, with end-point titres of 2-16. Of the 28 strains of V. cholerae non-O1 examined, 75%, 75%, 25% and 14.3% produced haemolysin that was active against erythrocytes of rabbit, sheep (Eltor haemolysin), chicken and man, respectively. Strains that produced a haemolysin active against both rabbit and sheep erythrocytes were dominant (35.7%). Ten (35.7%) of the 28 strains examined showed cell-associated haemagglutinating activity on human blood. Of the 10 strains, nine were isolated as sole pathogen and only one strain was associated with mixed infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411095 TI - Special issue. Red and white blood cell groups of chimpanzees: relationship to their human counterparts. PMID- 8411094 TI - Blood groups of primates: historical perspective and synopsis. PMID- 8411096 TI - The M-N--V-A-B-D blood group system of chimpanzee and other apes: serology and genetics. AB - Poly- and monoclonal anti-M and anti-N reagents detect on the red cells of anthropoid apes the M and/or N antigens which are similar to, but not identical with human M and N. A series of V-A-B-D specificities, closely related to the M-N system, are recognized on ape red blood cells by chimpanzee immune sera. To account for the distributions of the M-N--V-A-B-D types in man and in various apes, a genetic model is proposed that assumes the existence of two independent pairs of alleles: M/m, and N/n. In the processes of speciation, some of the alleles were lost or replaced by multiple mutations, resulting in chimpanzee in a series of codominant alleles responsible for as many as 16 M-N--V-A-B-D phenotypes. PMID- 8411097 TI - Recent advances in molecular and genetic analysis of Rh blood group structures. AB - The Rh antigens are carried by 30-32 kDa integral proteins of the red cell membrane. The Rh locus is composed of two genes: RhD, which encodes the major D antigen and is present only in Rh-positive genomes, and RhCcEe, which encodes both the Cc and Ee polypeptides, most likely by alternative splicing events. The D and non-D Rh mRNAs have been cloned. Their sequence homology suggest that the two Rh genes have evolved by duplication of a common ancestor. PMID- 8411098 TI - Application of maximum likelihood statistics for the population and family studies of inheritance of the chimpanzee R-C-E-F and V-A-B-D blood groups. AB - Maximum likelihood statistics were applied to the analysis of serological data to confirm the originally proposed genetic models of the chimpanzee R-C-E-F and V-A B-D systems. Five hundred ninety-nine chimpanzees, including 81 parents of 114 offspring, were tested for R-C-E-F, and 60 parents of 80 offspring were tested for V-A-B-D blood groups. An estimation-maximization procedure was used to obtain maximum likelihood estimates and support intervals of the haplotype frequencies. For each haplotype, the null hypothesis of nonexistence was evaluated. The frequencies obtained by this method do not differ significantly from those calculated by the square root formula, but put these estimates on a statistically more rigorous footing. PMID- 8411099 TI - Trans-specific Mhc polymorphism and the origin of species in primates. AB - The major histocompatibility complex (Mhc) is a cluster of loci controlling the specific immune response in vertebrates. Mhc alleles often differ by a large number of nucleotide substitutions, some of which began to accumulate before the emergence of extant species. We have applied the theory of allelic genealogy to the primate Mhc genes with the aim of estimating the size of the founding populations. The calculations indicate that the long-term effective population size of the studied species was between 10(4) and 10(5) individuals and that it most likely never dropped below 10(3) individuals. PMID- 8411100 TI - Primate genes for glycophorins carrying MN blood group antigens. AB - Glycophorin A, B, and E genes were derived from a common ancestral gene and this gene family appeared during primate evolution, probably between orangutan and gorilla divergences. Based on the study of genomic structures of these human glycophorins and the genetic and immunological study of primate glycophorins, we hypothesize that chimpanzee and gorilla glycophorin B could possess a longer extracellular region and carry a stronger N blood group antigenicity compared with that of the human. PMID- 8411101 TI - Whole inactivated SIV vaccine grown on human cells fails to protect against homologous SIV grown on simian cells. AB - Several groups have reported protection against experimental SIV infection in macaques immunized with a whole inactivated virus vaccine. The aim of the current study was to investigate whether five macaques vaccinated with whole inactivated SIV and previously shown to be protected against challenge with two divergent strains of SIV grown on human cells could resist challenge with a subsequent homologous SIV grown on macaque cells. We show here that this same vaccine did not protect when the challenge virus was grown on primary cells of monkey origin. PMID- 8411102 TI - Immunisation of macaques with SIV env recombinants: specificity of T cell and antibody responses and evaluation of protective efficacy. AB - Macaques were immunised with lentil lectin purified recombinant SIVmac (BK28) derived gp160 (rgp160) with or without live vaccinia (vac)-env (BK28) priming, followed by a final boost with solid matrix antibody antigen (SMAA)-gp160 (J5) complexes and challenged with the SIVmac molecularly cloned virus J5M. Rgp160 and vac-env plus gp160 induced strong Ab responses against the homologous virus. Live vac-env did not enhance or prolong the antibody response, however, T cell responses were stronger. Analysis of the specificity of the immune response demonstrated that sequence variation within SIVmac viruses can affect antibody and T cell recognition. A single booster immunisation with the heterologous SIVmac J5 env recombinant protein was not sufficient to protect against the molecularly cloned virus J5M. These findings further illustrate the difficulty of generating protective immunity with immunogens based on single sequence recombinants. PMID- 8411103 TI - Immunization of Macaca fascicularis with inactivated SIV preparations: challenge with human- or monkey-derived SIV and the effects of a longer immunization schedule. AB - In cynomolgus monkeys, we compared two human-derived SIVmac251 whole virus vaccines, a long vs short immunization schedule, and two different challenge viruses. Both vaccines induced protection after challenge with human-derived SIVmac251/32H. There was no difference between the two schedules of immunization. Seven monkeys, five of which were protected following the first challenge, were reboosted and rechallenged with monkey-derived SIVmac251, but no protection was observed. The titers of anti-human cell or -SIV neutralizing antibodies were not related to protection. PMID- 8411104 TI - Detection of serum antibodies in Ethiopian baboons that cross-react with SIV, HTLV-I, and type D retroviral antigens. AB - Baboons (Papio cynocephalus) imported from Ethiopia were screened for antibodies to various primate retroviruses by immunoblotting. Antibodies that cross-reacted with SIV/Mne or with type D viral antigens were detected in approximately one third of these animals. In addition, 20% of these baboons had antibodies that cross-reacted with HTLV-I viral antigens. These data suggest that wild-caught baboons are infected with retroviruses only partially related to known primate viral isolates. PMID- 8411105 TI - Dynamics of the immune system response in cerebrospinal fluid and blood of SIVmac infected rhesus monkeys. AB - Paired sera and CSF samples were collected from SIVmac-infected macaques. Animals infected with SIVmac251 maintained low gag and high env-specific antibody levels in plasma. Increasing env-specific antibody titers in CSF were associated in one animal with strong intrathecal synthesis. SIVmac239-infected monkeys revealed high antibody titers of gag and env-specificity, in one animal accompanied by weak intrathecal synthesis of virus-specific antibodies. In all animals, the CD4/CD8 ratio in CSF decreased faster compared to blood. PMID- 8411106 TI - The combined assessment of cellular apoptosis, mitochondrial function and proliferative response to pokeweed mitogen has prognostic value in SIV infection. AB - Using the assessment of the mitochondrial metabolic activity of freshly isolated blood mononuclear cells, the flow cytometric detection of apoptosis and of the proliferative responses to PWM, SIV-infected macaques were classified in: stage 0, which included all animals with unaffected parameters, and stages 1, 2, and 3, which included animals having one, two, or all three parameters affected, respectively. This novel three-parametric staging system (ISS) provides a new prognostic tool in the longitudinal study of SIV infection. PMID- 8411107 TI - Pathogenesis of SIVmac251 after atraumatic inoculation of the rectal mucosa in rhesus monkeys. AB - Intrarectal inoculation of rhesus monkeys with low doses of SIVmac led to a prolonged clinical and virological latency that was not observed for high intrarectal doses or for intravenous inoculation. Animals infected intrarectally with low virus doses remained negative for serum antibody responses to SIV for at least one year even though they readily transferred SIV to naive recipients via transfusion of whole blood. PMID- 8411108 TI - Intra-amniotic inoculation of pigtailed macaque (Macaca nemestrina) fetuses with SIV and HIV-1. AB - Six pregnant pigtailed macaques (Macaca nemestrina) were inoculated intra amniotically (i.a.) with SIVMne. All became viremic and seroconverted; three viable offspring were SIV-positive and at autopsy showed disseminated viral infection; one of three abortuses had SIV-infected thymic macrophages. Three of five pregnant macaques inoculated i.v. and/or i.a. with HIV-1LAI became virus positive, and four seroconverted, suggesting fetal-maternal transmission. One abortus had HIV-1-antigen in lymph nodes and brain; one infant, culture-positive at birth, died at age 11 days of disseminated HIV-1 infection. PMID- 8411109 TI - Distribution of SIV infection in the gastrointestinal tract of rhesus macaques at early and terminal stages of AIDS. AB - Intestinal tissues from SIV-infected rhesus macaques with subclinical as well as with terminal SIV disease were evaluated for SIV infection. SIV-infected mononuclear cells were detected throughout the gastrointestinal tract in all animals. In early infection, SIV-infected cells were diffusely distributed in lymphoid tissue and lamina propria, whereas in late stages of the disease, the viral infection was more severe and widely disseminated. Lymphoid cells of the lamina propria and gut-associated lymphoid tissue were the primary targets of SIV. PMID- 8411110 TI - First updated and revised survey of worldwide HIV and SIV vaccine challenge studies in nonhuman primates: progress in first and second order studies. PMID- 8411111 TI - Long-term protection of macaques against high-dose type D retrovirus challenge after immunization with recombinant vaccinia virus expressing envelope glycoproteins. AB - Recombinant vaccinia virus expressing the envelope proteins of type D retrovirus Washington (SRV-2/W) was used to immunize macaques against SRV-2 infection. Four immunized macaques which had resisted a prior low-dose challenge were rechallenged with a high dose (10(6) infectious particles) of SRV-2 two years after being immunized. All four non-immunized control macaques became infected, but the four vaccinated animals resisted this intravenous challenge, as determined by the inability to detect SRV-2 in peripheral blood mononuclear cells and by the lack of seroconversion to new viral antigens. PMID- 8411112 TI - The importance of nonhuman primate research in the battle against AIDS: a historical perspective. PMID- 8411113 TI - Protection of vaccinia-primed macaques against SIVmne infection by combination immunization with recombinant vaccinia virus and SIVmne gp160. AB - Two Macaca fascicularis with preexisting immunity to vaccinia virus were immunized twice with recombinant vaccinia virus expressing SIVmne gp160. Their SIV-specific antibody responses were lower than that of vaccinia-naive animals immunized similarly. Upon repeated boosting with gp160, the SIV-specific antibody titers in vaccinia-primed animals reached similar levels as vaccinia-naive animals and with comparable neutralizing titers. Both animals were protected against repeated intravenous challenge with low-dose SIVmne E11S. These results are significant because SIVmne E11S infection in M. fascicularis is pathogenic and leads to AIDS-like diseases. PMID- 8411114 TI - The alpha 1 Na(+)-K+ pump of the Dahl salt-sensitive rat exhibits altered Na+ modulation of K+ transport in red blood cells. AB - The properties of the alpha 1 Na(+)-K+ pump were compared in Dahl salt-sensitive (DS) and salt-resistant (DR) strains by measuring ouabain-sensitive fluxes (mmol/liter cell x hr = FU, Mean +/- SE) in red blood cells (RBCs) and varying internal (i) and external (o) Na+ and K+ concentrations. Kinetic parameters of several modes of operation, i.e., Na+/K+, K+/K+, Na+/Na+ exchanges, were characterized and analyzed for curve-fitting using the Enzfitter computer program. In unidirectional flux studies (n = 12 rats of each strain) into fresh cells incubated in 140 mM Na(+) + 5 mM K+, ouabain-sensitive K+ influx was substantially lower in the DS than in DR RBCs, while ouabain-sensitive Na+ efflux and Nai were similar in both strains. Thus, the coupling ratio between unidirectional Na+:K+ fluxes was significantly higher in DS than in DR cells at similar RBC Na+ content. In the presence of 140 mM Nao, activation of ouabain sensitive K+ influx by Ko had a lower Km and Vmax in DS as estimated by the Garay equation (N = 2.70 +/- 0.33, Km 0.74 +/- 0.09 mM; Vmax 2.87 +/- 0.09 FU) than in DR rats (N = 1.23 +/- 0.36, Km 2.31 +/- 0.16 mM; Vmax 5.70 +/- 0.52 FU). However, the two kinetic parameters were similar following Nao removal. The activation of ouabain-sensitive K+ influx by Nai had significantly lower Vmax in DS (9.3 +/- 0.4 FU) than in DR (14.5 +/- 0.6 FU) RBCs but similar Km. These data suggest that the low K+ influx in DS cells is caused by a defect in modulation by Nao and Nai. Na+ efflux showed no differences in Nai activation or trans effects by Nao and Ko, thus accounting for the different Na+:K+ coupling ratio in the Dahl strains. Further evidence for the differences in the coupling of ouabain-sensitive fluxes was found in studies of net Na+ and K+ fluxes, where the net ouabain-sensitive Na+ losses showed similar magnitudes in the two Dahl strains while the net ouabain-sensitive K+ gains were significantly greater in the DR than the DS RBCs. Ouabain-sensitive Na+ influx and K+ efflux were also measured in these rat RBCs. The inhibition of ouabain-sensitive Na+ influx by Ko was fully competitive for the DS but not for the DR pumps. Thus, for DR pumps, Ko could activate higher K+ influx in DR pumps without a complete inhibition of ouabain-sensitive Na+ influx.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411115 TI - Blockade of a KCa channel with synthetic peptides from noxiustoxin: a K+ channel blocker. AB - Using the outside-out configuration of the patch-clamp method, we studied the effect of several synthetic peptides corresponding to various segments from the N terminal region of noxiustoxin (NTX) on single Ca(2+)-activated K+ (KCa) channels of small conductance obtained from cultured bovine aortic endothelial cells. These peptides induced diverse degrees of fast blockade in the endothelial KCa channel. The most effective blockers were the peptides NTX1-39 (IC50 = 0.5 microM) and NTX1-20 comprising the first 20 amino acids from the native toxin (IC50 approximately 5 microM), while less effective was the hexapeptide NTX1-6, from the first six amino acid residues of NTX (IC50 = 500 microM). This was the minimum sequence required to block the channel. By testing overlapping sequences from the entire molecule, specially those corresponding to the N-terminal region of NTX, we have been able to determine their different apparent affinities for the KCa channel. Synthetic peptides from the C-terminal region produced no effect on the KCa channel at the concentrations tested (up to 1 mM). These results confirm that in the N-terminal region of the NTX is located part of the sequence that may recognize K+ channels, as we have suggested previously from in vivo experiments. The blockade induced by native NTX was poorly affected by changes in membrane potential; however, the blockage induced by synthetic peptides lacking the C-terminal region was partially released by depolarization. PMID- 8411116 TI - ATP-inhibited and Ca(2+)-dependent K+ channels in the soma membrane of cultured leech Retzius neurons. AB - The properties of one ATP-inhibited and one Ca(2+)-dependent K+ channel were investigated by the patch-clamp technique in the soma membrane of leech Retzius neurons in primary culture. Both channels rectify at negative potentials. The ATP inhibited K+ channel with a mean conductance of 112 pS is reversibly blocked by ATP (Ki = 100 microM), TEA (Ki = 0.8 mM) and 10 mM Ba2+ and irreversibly blocked by 10 nM glibenclamide and 10 microM tolbutamide. It is Ca2+ and voltage independent. Its open state probability (Po) decreases significantly when the pH at the cytoplasmic face of inside-out patches is altered from physiological to acid pH values. The Ca(2+)-dependent K+ channel with a mean conductance of 114 pS shows a bell-shaped Ca2+ dependence of Po with a maximum at pCa 7-8 at the cytoplasmic face of the membrane. The Po is voltage independent at the physiologically relevant V range. Ba2+ (10 MM) reduces the single channel amplitude by around 25% (ATP, TEA, glibenclamide, tolbutamide, and Ba2+ were applied to the cytoplasmic face of the membrane). We conclude that the ATP dependent K+ channel may play a role in maintaining the membrane potential constant--independently from the energy state of the cell. The Ca(2+)-dependent K+ channel may play a role in generating the resting membrane potential of leech Retzius neurons as it shows maximum activity at the physiological intracellular Ca2+ concentration. PMID- 8411117 TI - Rapid increase in pH set-point of the Na(+)-in-dependent chloride/bicarbonate antiporter in Vero cells exposed to heat shock. AB - Internal pH (pHi) is in Vero cells regulated mainly by three antiports. Na+/H+ antiport and Na(+)-dependent Cl-/HCO3- antiport increase pHi in acidified cells, and Na(+)-independent Cl-/HCO3- antiport lowers pHi in cells after alkalinization. The activities of the antiporters were altered in cells after exposure to 41-45 degrees C. Under such conditions the Na+/H+ antiport and the Na(+)-dependent Cl-/HCO3- antiport were both stimulated, whereas the Na(+) independent Cl-/HCO3- antiport was inhibited in such a way that a higher pH value was required to activate it. This alteration was also induced by some other forms of cellular stress, but did most likely not involve stress proteins as protein synthesis was not required. The possibility of regulation by alteration in protein phosphorylation is discussed. PMID- 8411118 TI - Voltage-dependent calcium and potassium conductances in striated muscle fibers from the scorpion, Centruroides sculpturatus. AB - Ionic currents responsible for the action potential in scorpion muscle fibers were characterized using a three-intracellular microelectrode voltage clamp applied at the fiber ends (8-12 degrees C). Large calcium currents (ICa) trigger contractile activation in physiological saline (5 mM Ca) but can be studied in the absence of contractile activation in a low Ca saline (< or = 2.5 mM). Barium (Ba) ions (1.5-3 mM) support inward current but not contractile activation. Ca conductance kinetics are fast (time constant of 3 msec at 0 mV) and very voltage dependent, with steady-state conductance increasing e-fold in approximately 4 mV. Half-activation occurs at -25 mV. Neither ICa nor IBa show rapid inactivation, but a slow, voltage-dependent inactivation eliminates ICa at voltages positive to -40 mV. Kinetically, scorpion channels are more similar to L-type Ca channels in vertebrate cardiac muscle than to those in skeletal muscle. Outward K currents turn on more slowly and with a longer delay than do Ca currents, and K conductance rises less steeply with voltage (e-fold change in 10 mV; half-maximal level at 0 mV). K channels are blocked by externally applied tetraethylammonium and 3,4 diaminopyridine. PMID- 8411119 TI - Whole cell K and Cl currents in dissociated eccrine secretory coil cells during stimulation. AB - Using the whole-cell voltage clamp (to determine the membrane current) and current clamp (to determine membrane potential) methods in conjunction with the nystatin-perforation technique, we studied the effect of methacholine (MCh) and other secretagogues on whole cell K and Cl currents in dissociated rhesus palm eccrine sweat clear cells. Application of MCh by local superfusion induced a net outward current (at a holding potential of -60 mV and a clamp voltage of 0 mV), and a transient hyperpolarization by 5.6 mV, suggesting the stimulation of K currents. The net outward current gradually changed to the inward (presumably Cl) currents over the next 1 to 2 min of continuous MCh stimulation. During this time the membrane potential also changed from hyperpolarization to depolarization. The inward currents were increasingly more activated than outward (presumably K) currents during repeated MCh stimulations so that a net inward current (at -60 mV) was observed after the fourth or fifth MCh stimulation. Ionomycin (10 microM) also activated both inward and outward current. The observed effect of MCh was abolished by reducing extracellular [Ca] to below 1 nM (Ca-free + 1 mM EGTA in the bath). MCh-activated outward currents were inhibited by 5 mM Ba and by 0.1 mM quinidine, although these agents also suppressed the inward currents. Bi-ionic potential measurements indicated that the contribution of Na to the membrane potential was negligible both before and after MCh or ISO (isoproterenol) stimulations and that the observed membrane current was carried mainly by K and Cl. MCh increased the bi-ionic potential by step changes in external K and Cl concentrations, further supporting that MCh-induced outward and inward currents represent K and Cl currents, respectively. Stimulation with ISO or FK (forskolin) resulted in a depolarization by about 55 mV and a net inward (most likely Cl) current independent of external Ca. CT-cAMP mimicked the effects of FK and ISO. The bi-ionic potential, produced by step changes in the external Cl concentration, increased during ISO stimulation, whereas that of K decreased. This indicates that the ISO-induced inward current is due to Cl current and that K currents were unchanged or slightly decreased during stimulation with ISO or 10 microM FK. Both myoepithelial and dark cells responded only to MCh (but not to FK) with a marked depolarization of the membrane potential due to activation of Cl, but not K, currents. We conclude that MCh stimulates Ca-dependent K and Cl currents, whereas ISO stimulates cAMP-dependent Cl currents in eccrine clear cells. PMID- 8411120 TI - Interaction between red cell membrane band 3 and cytosolic carbonic anhydrase. AB - We have previously proposed that a membrane transport complex, centered on the human red cell anion transport protein, band 3, links the transport of anions, cations and glucose. Since band 3 is specialized for HCO3-/Cl- exchange, we thought there might also be a linkage with carbonic anhydrase (CA) which hydrates CO2 to HCO3-. CA is a cytosolic enzyme which is not present in the red cell membrane. The rate of reaction of CA with the fluorescent inhibitor, dansylsulfonamide (DNSA) can be measured by stopped-flow spectrofluorimetry and used to characterize the normal CA configuration. If a perturbation applied to a membrane protein alters DNSA/CA binding kinetics, we conclude that the perturbation has changed the CA configuration by either direct or allosteric means. Our experiments show that covalent reaction of the specific stilbene anion exchange inhibitor, DIDS, with the red cell membrane, significantly alters DNSA/CA binding kinetics. Another specific anion exchange inhibitor, benzene sulfonate (BSate), which has been shown to bind to the DIDS site causes a larger change in DNSA/CA binding kinetics; DIDS reverses the BSate effect. These experiments show that there is a linkage between band 3 and CA, consistent with CA interaction with the cytosolic pole of band 3. PMID- 8411121 TI - Tolbutamide-sensitive potassium conductance in the basolateral membrane of A6 cells. AB - K+ channels sensitive to intracellular ATP (KATP channels) have been described in a number of cell types and are selectively inhibited by sulfonylurea drugs. To look for the presence of this type of K+ channel in the basolateral membrane of tight epithelia, we have used an amphibian renal cell line, the A6 cells, grown on filters. After the selective permeabilization of the apical membrane with amphotericin B, the basolateral conductance was studied under voltage-clamp conditions. Tolbutamide inhibited 65.8 +/- 6.3% of the barium-sensitive current. The tolbutamide-sensitive conductance had an equilibrium potential of -83 +/- 1 mV and was inward rectifying in spite of the outwardly directed K+ gradient. Similar results were obtained with glibenclamide. The half-inhibition constants were 25.7 +/- 3.0 microM and 0.114 +/- 0.018 microM for tolbutamide and glibenclamide, respectively. To study the relation between cellular ATP and the activity of this conductance, A6 cells were treated with glucose (5 mM) and insulin (250 microU/ml). This maneuver significantly increased the cellular ATP and abolished the tolbutamide-sensitive conductance. A sulfonylurea-sensitive K+ conductance is present and active in the basolateral membrane of A6 cells. This conductance appears to be modulated by physiological changes of intracellular ATP. PMID- 8411122 TI - Characterization of dietary phosphorus-dependent duodenal calcium uptake in vitamin D-deficient chicks. AB - The effect of dietary phosphorus on intestinal calcium uptake was examined in duodenal cells isolated from vitamin D-deficient chicks. Cells from chicks on a high phosphorus diet accumulated calcium at a rate 38% higher than cells from animals on a normal phosphorus diet. Diet high in calcium did not affect calcium absorption in duodenal cells. The dietary phosphorus effect on calcium absorption was specific. Uptake of alpha-methyl glucose was not altered. Increase in calcium absorption by a high phosphorus diet was not due to a change in cellular energy metabolism nor to the content of phosphorus in cells. Kinetically, a high phosphorus diet increased the Vmax of calcium uptake; the affinity for calcium was unaffected. The effectiveness of dietary phosphorus to enhance the intestinal calcium uptake could also be demonstrated in brush border membrane vesicles. The increase in calcium uptake was not due to an alteration in membrane binding capacity nor to calcium efflux from vesicles. To test the hypothesis that a high phosphorus diet may affect membrane transport by altering phospholipid metabolism in duodenal cells, we examined the phospholipid content in isolated brush border membranes. The content of phosphatidylcholine, phosphatidylserine, phosphatidylinositol and phosphatidylethanolamine was not altered by the high phosphorus diet. These findings suggest that the vitamin D-independent and dietary phosphorus-dependent effect on intestinal calcium absorption was primarily due to a change in the calcium flux at the luminal side of the cells. However, the precise mechanism is still not clear. PMID- 8411123 TI - Platelet activating factor-induced increase in cytosolic calcium and transmembrane current in human macrophages. AB - Platelet-activating factor (PAF) is synthesized and secreted by macrophages in responding to inflammatory stimuli. When exogenously applied to human monocyte derived macrophages (HMDMs), PAF induces a rapid rise in cytosolic free calcium (Cai) believed to be an early triggering event in macrophage activation. We investigated PAF-induced Ca2+ signaling in HMDMs using the calcium indicator Fura 2, combining single cell ratio fluorimetry and digital video imaging with whole cell recording techniques. Application of PAF (20 ng/ml) to adherent macrophages induced transient increases in Cai that were biphasic, consisting of an initial phase that could be observed in Ca(2+)-free solutions and a second phase that was critically dependent upon Ca2+ entry. When Mn2+ was applied to cells in the presence and absence of Ca2+, PAF increased the rate of Mn2+ entry rate only when Ca2+ was absent. PAF increased the rate of Ba2+ entry even when measured in the presence of external Ca2+. Ca2+ entry was reversibly inhibited in the presence of external La3+ (1 mM). Data obtained from simultaneous voltage clamp/microfluorimetry experiments demonstrated the activation of a nonselective cation current which closely paralleled the rising phase of the Cai transient. We investigated whether the non-selective cation conductance provided for the bulk of the agonist-induced Ca2+ influx. Changes in Cai following removal of extracellular Ca2+ (Cao) during the agonist-induced Cai response were not associated with changes in whole-cell current. The inability to detect whole-cell current changes correlated with a decrease in Cao suggests that the bulk of the Ca2+ influx was not through the nonselective conductance and either does not occur through a conductance pathway or occurs via a parallel pathway consisting of channels which are both low conductance and highly Ca2+ selective. PMID- 8411124 TI - Analyzing an electrogenic cotransporter. AB - A model for the sodium-dependent accumulation of glutamate by synaptosomes has been presented which fits the data of Wheeler and his coworkers and supports their hypothesis of an electrogenic cotransporter. Since their hypothesis was based on experimental data on the operation of the cotransporter on the outer membrane, the model was expanded to predict events when the cotransporter was operating on both sides of the membrane. The model predicts that the accumulation of glutamate is sensitive to the synaptosomal sodium and emphasizes the importance of the sodium/potassium pump to maintain this value. A model which uses only an electrogenic form of the cotransporter on the external membrane and a neutral form on the inside of the membrane predicts too much or too little accumulation of glutamate at different membrane potentials. A model which uses an electrogenic cotransporter on the external membrane and a concentration-dependent sodium glutamate leak would require a significant increase in the permeability of sodium glutamate when the membrane depolarizes. Only the operation of all four mentioned mechanisms will fit experimental data at two different external sodium concentrations and over the range of membrane potentials measured experimentally. PMID- 8411125 TI - Responsiveness of cardiac Na+ channels to a site-directed antiserum against the cytosolic linker between domains III and IV and their sensitivity to other modifying agents. AB - Elementary Na+ currents were recorded in inside-out patches from neonatal rat heart cardiocytes to analyze the influence of a site-directed polyclonal anti serum against the linker region between the domains III and IV (amino acids 1489 1507 of the cardiac Na+ channel protein) on Na+ channel gating and to test whether this part of the alpha-subunit may be considered as a target for modifying agents such as the (-)-enantiomer of DPI 201-106. Anti-SLP 1 serum (directed against amino acids 1490-1507) evoked, usually within 10-15 min after cytosolic administration, modified Na+ channel activity. Antiserum-modified Na+ channels retain a single open state but leave, at -60 mV for example, their conducting configuration consistently with an about threefold lower rate than normal Na+ channels. Another outstanding property of noninactivating Na+ channels, enhanced burst activity, may be quite individually pronounced, a surprising result which is difficult to interpret in terms of structure-function relations. Removal of inactivation led to an increase of reconstructed peak INa (indicating a rise in NPo) and changed INa decay to obey second-order kinetics, i.e., open probability declined slowly but progressively during membrane depolarization. The underlying deactivation process is voltage dependent and responds to a positive voltage shift with a deceleration but may operate even at the same membrane potential with different rates. Iodate-modified Na+ channels exhibit very similar properties including a conserved conductance. They are likewise controlled by an efficient, voltage-dependent deactivation process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411126 TI - Independence of water and solute pathways in human RBCs. AB - We have investigated the permeability of the human red blood cell to four di hydroxy alcohols, 1,2PD (1,2 propanediol), 1,3PD (1.3 propanediol), 1,4BD (1,4 butanediol), and 2,3BD (2,3 butanediol), and to water by using a recently developed ESR stopped-flow method which is free from artifacts found in light scattering methods. Numerical solutions of the Kedem-Katchalsky equations fit to experimental data yielded the following permeability coefficients: P1,2PD = 3.17 x 10(-5) cm sec-1, P1,3PD = 1.75 x 10(-5) cm sec-1, P1,4BD = 2.05 x 10(-5) cm sec 1, P2,3BD = 7.32 x 10(-5) cm sec-1. Reflection coefficients (sigma) were evaluated by comparing data fit with assumed values of sigma = 0.6, 0.8 and 1.0. In all four cases the best fit was obtained with sigma = 1.0. Treatment of cells with PCMBS (para-chloro mercuri-benzene-sulfonate) was followed by a large (> 10 fold) decrease in water permeability with virtually no change in alcohol permeability. We conclude that these alcohols do not permeate the water channels to any significant extent, and discuss some of the problems in light scattering measurements of reflection coefficients that could lead to erroneous values for sigma. PMID- 8411127 TI - Glycine uptake by trout (Salmo trutta) red blood cells. AB - The present study demonstrates the presence of different amino acid carriers in the membrane of trout red cells. Most glycine is taken up through the Na(+) dependent system ASC, although the nearly specific Gly system is also active. Besides these carriers, glycine is taken up by means of Na(+)-independent transporters, system L being the most important. A system asc of high affinity and low capacity has been found, and band 3 is unable to transport glycine under physiological conditions. These results suggest that although all these carriers are already present in primitive vertebrates, several differences exist in their properties with respect to those found in mammalian cells. PMID- 8411128 TI - Metabolic regulation of glucose transport. PMID- 8411129 TI - Necturus gallbladder epithelial cell volume regulation and inhibitors of arachidonic acid metabolism. AB - Inhibition of the metabolism of arachidonic acid by the epoxygenase (cytochrome P 450) pathway with the inhibitor ketoconazole results in excessive cell swelling upon exposure to hyposmolality instead of the rapid and complete regulatory volume decrease (RVD) normally observed. NaCl entry from bathing solutions to cell interior was shown to cause this swelling, with Na influx occurring across the basolateral membrane and electrically silent Cl influx across the apical membrane. Ion substitution experiments show that the KCl efflux mediating RVD was unimpaired by ketoconazole, but was overwhelmed by the NaCl influx. Measurements of transepithelial fluid flux, Cl concentration, osmolality and pH showed that gallbladders treated with ketoconazole transiently secreted fluid rather than the normal absorption. We conclude that inhibition of arachidonic acid metabolism does not directly affect RVD by Necturus gallbladder, but that blockade of the epoxygenase pathway can have a profound influence on NaCl entry into gallbladder epithelial cells. PMID- 8411130 TI - Effect of Nai on activity and voltage dependence of the Na/K pump in adult rat cardiac myocytes. AB - We have measured the voltage dependence of the Na/K pump in isolated adult rat cardiac myocytes using the whole-cell patch-clamp technique. In the presence of 1 2 mM Ba and 0.1 mM Cd and nominally Ca-free, Na/K pump current (Ip) was measured as the change in current due to 1 mM ouabain. Voltage dependence of Ip was measured between -140 and +40 or +60 mV using square voltage-pulse and voltage ramp protocols, respectively. With 150 mM extracellular Na (Nao) and 5.4 mM extracellular K (Ko), we found that the Na/K pump shows a strong positive voltage dependence between -140 and 0 mV and is voltage independent at positive potentials. Removing Nao reduced the voltage dependence at negative potentials with no effect at positive potentials. When Ko was reduced, a negative slope appeared in the current-voltage (I-V) curve at positive potentials. We have investigated whether Nai (intracellular Na) might also affect the voltage dependence of Ip by varying Na in the patch pipette (Napip) between 20 and 85 mM. We found, as expected, that Ip increased markedly as Napip was raised, saturating at about 70 mM Napip under these conditions. In contrast, while Ip saturated near +20 mV and declined to about 40% of maximum at -120 mV, there was no effect of Napip under these conditions. In contrast, while Ip saturated near +20 mV and declined to about 40% of maximum at -120 mV, there was no effect of Napip on the voltage dependence of Ip. This suggests that neither Nai binding to the Na/K pump nor the conformational changes dependent on Nai binding are voltage dependent. These results are consistent with extracellular ion binding within the field of the membrane but do not rule out the possibility that other steps, such as Na translocation, are also voltage dependent. PMID- 8411131 TI - Cytoplasmic Ca2+ does not inhibit the cardiac muscle sarcoplasmic reticulum ryanodine receptor Ca2+ channel, although Ca(2+)-induced Ca2+ inactivation of Ca2+ release is observed in native vesicles. AB - Single channel properties of cardiac and fast-twitch skeletal muscle sarcoplasmic reticulum (SR) release channels were compared in a planar bilayer by fusing SR membranes in a Cs(+)-conducting medium. We found that the pharmacology, Cs+ conductance and selectivity to monovalent and divalent cations of the two channels were similar. The cardiac SR channel exhibited multiple kinetic states. The open and closed lifetimes were not altered from a range of 10(-7) to 10(-3) M Ca2+, but the proportion of closed and open states shifted to shorter closings and openings, respectively. However, while the single channel activity of the skeletal SR channel was activated and inactivated by micromolar and millimolar Ca2+, respectively, the cardiac SR channel remained activated in the presence of high [Ca2+]. In correlation to these studies, [3H]ryanodine binding by the receptors of the two channel receptors was inhibited by high [Ca2+] in skeletal but not in cardiac membranes in the presence of adenine nucleotides. There is, however, a minor inhibition of [3H]ryanodine binding of cardiac SR at millimolar Ca2+ in the absence of adenine nucleotides. When Ca(2+)-induced Ca2+ release was examined from preloaded native SR vesicles, the release rates followed a normal biphasic curve, with Ca(2+)-induced inactivation at high [Ca2+] for both cardiac and skeletal SR. Our data suggest that the molecular basis of regulation of the SR Ca2+ release channel in cardiac and skeletal muscle is different, and that the cardiac SR channel isoform lacks a Ca(2+)-inactivated site. PMID- 8411132 TI - Lateral mobility of lipid analogues and GPI-anchored proteins in supported bilayers determined by fluorescent bead tracking. AB - Lipid analogues and glycosylphosphatidylinositol (GPI)-anchored proteins incorporated in glass-supported phospholipid bilayers (SBL) were coupled to small (30 nm diameter) fluorescent beads whose motion in the liquid phase was tracked by intensified fluorescence video microscopy. Streptavidin (St), covalently attached to the carboxyl modified surface of the polystyrene bead, bound either the biotinylated membrane component, or a biotinylated monoclonal antibody (mAb) directed against a specific membrane constituent. The positions of the beads tethered to randomly diffusing membrane molecules were recorded at 0.2 sec intervals for about 1 min. The mean square displacement (rho) of the beads was found to be a linear function of diffusion time t, and the diffusion coefficient, D, was derived from the relation, rho(t) = 4Dt. The values of D for biotinylated phosphatidylethanolamine (Bi-PE) dispersed in an egg lecithin:cholesterol (80:20%) bilayer obtained by this methodology range from 0.05 to 0.6 micron 2/sec with an average of mean value of D = 0.26 micron 2/sec, similar to the value of mean value of D = 0.24 micron 2/sec for fluorescein-conjugated phosphatidylethanolamine (Fl-PE) linked to St-coupled beads by the anti fluorescein mAb 4-4-20 or its Fab fragment. These values of D are comparable to those reported for Fl-PE linked to 30 nm gold particles but are several times lower than that of Fl-PE in the same planar bilayer as measured by fluorescence photobleaching recovery, D = 1.3 microns 2/sec. The mobilities of two GPI anchored proteins in similar SBL were also determined by use of the appropriate biotinylated mAb and were found to be mean value of D = 0.25 and 0.56 micron 2/sec for the decay accelerating factor (DAF, CD55) and the human Fc gamma RIIIB (CD16) receptors, respectively. The methodology described here is suitable for tracking any accessible membrane component. PMID- 8411133 TI - Comparison of lowest energy conformations of dimethylcurine and methoxyverapamil: evidence of ternary association of calcium channel, Ca2+, and calcium entry blockers. AB - Verapamil and dimethylcurine are Ca2+ entry blockers of essentially different chemical structures which presumably bind to the same arylalkylamine receptor of the L-type Ca channel. A systematic conformational analysis of methoxyverapamil (D-600) and dimethylcurine has been carried out using a molecular mechanics method. The lowest minimum-energy conformations of D-600 are predisposed to chelate Ca2+ by four oxygen atoms of the stacked methoxyphenyl moieties. Comparison of the lowest energy conformations of D-600-Ca2+ and dimethylcurine revealed a similar spatial disposition of cationic groups and methoxyphenyl moieties in the two compounds. A three-dimensional model of arylalkylamine receptor was suggested which incorporates two nucleophilic areas of the Ca channel. Dimethylcurine binds to these areas by its quaternary amine functions, whereas D-600 does so by amine function and via coordinated Ca2+. The results support the hypotheses on ternary complex formation between the ligands of Ca channel, their receptors, and Ca2+. PMID- 8411134 TI - Basolateral Cl/HCO3 exchange in rat jejunum: the effect of sodium. AB - Basolateral membrane vesicles isolated from rat jejunum were used to characterize a Cl/HCO3 exchange mechanism previously evidenced. Cl uptake experiments provided no evidence for Cl/OH countertransport, confirming anyhow the presence of Cl/HCO3 antiport, which was inhibited by 2 mM furosemide and unaffected by 2 mM amiloride. An outwardly directed Na gradient stimulated Cl uptake and this effect was increased if Na was present at both vesicle surfaces. To investigate the mechanism of coupling between Na and the transport protein, we performed Na uptake experiments. Na uptake was unaffected by cis-bicarbonate and trans-Cl gradients; the reversal of anion gradients was still ineffective. Similar results were obtained when a pH difference across the membrane vesicles was imposed. This study seems to suggest that Na is not transported by the Cl/HCO3 exchanger and that another mode of Na dependence must be taken into account. PMID- 8411135 TI - Hepatocyte water volume and potassium activity during hypotonic stress. AB - Hepatocytes exhibit a regulatory volume decrease (RVD) during hypotonic shock, which comprises loss of intracellular K+ and Cl- accompanied by hyperpolarization of transmembrane potential (Vm) due to an increase in membrane K+ conductance, (GK). To examine hepatocyte K+ homeostasis during RVD, double-barrel, K+ selective microelectrodes were used to measure changes in steady-state intracellular K+ activity (aKi) and Vm during hyposmotic stress. Cell water volume change was evaluated by measuring changes in intracellular tetramethylammonium (TMA+). Liver slices were superfused with modified Krebs physiological salt solution. Hyposmolality (0.8 x 300 mosm) was created by a 50 mM step-decrease of external sucrose concentration. Hepatocyte Vm hyperpolarized by 19 mV from -27 +/- 1 to -46 +/- 1 mV and aiK decreased by 14% from 91 +/- 4 to 78 +/- 4 mM when slices were exposed to hyposmotic stress for 4-5 min. Both Vm and aKi returned to control level after restoring isosmotic solution. In paired measurements, hypotonic stress induced similar changes in Vm and aKi in both control and added ouabain (1 mM) conditions, and these values returned to their control level after the osmotic stress. In another paired measurement, hypotonic shock first induced an 18-mV increase in Vm and a 15% decrease in aKi in control condition. After loading hepatocytes with TMA+, the same hypotonic shock induced a 14-mV increase in Vm and a 14% decrease in aTMAi. This accounted for a 17% increase of intracellular water volume, which was identical to the cell water volume change obtained when aKi was used as the marker.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411136 TI - Cl- channels in basolateral renal medullary membranes: VII. Characterization of the intracellular anion binding sites. AB - A unique property of basolateral membrane Cl- channels from the mTAL is that the Cl- concentration facing the intracellular aspects of these channels is a determinant of channel open time probability (Po). The K1/2 for maximal activation of Po by Cl- facing intracellular domains of these channels is 10 mM Cl-. The present experiments evaluated the nature of these Cl(-)-interactive sites. First, we found that the impermeant anion isethionate, when exposed to intracellular Cl- channel faces, could augment Po with a K1/2 in the range of 10 mM isethionate without affecting conductance (gCl, pS). Second, pretreatment of the solutions facing the intracellular aspects of the channels with either 1 mM phenylglyoxal (PGO), an arginine-specific reagent, or the lysine/terminal amine reagent trinitrobenzene sulfonic acid (TNBS, 1 mM), prevented the activation of Po usually seen when the Cl- concentration of solutions facing intracellular channel domains was raised from 2 to 50 mM. However, when the Cl- channel activity was increased by first raising the Cl- concentration bathing intracellular channel faces from 2 to 50 mM, subsequent addition of either PGO or TNBS to solutions bathing intracellular Cl- channel faces had no effect on Po. We conclude that the intracellular aspects of these Cl- channels contain Cl(-) interactive loci (termed [Cl]i) which are accessible to impermeant anions in intracellular fluids and which contain arginine- and lysine-rich domains which can be inactivated, at low ambient Cl- or isethionate concentrations, by interactions with PGO or TNBS. PMID- 8411137 TI - Double whole-cell patch-clamp characterization of gap junctional channels in isolated insect epidermal cell pairs. AB - Double whole-cell patch-clamp methods were used to characterize junctional membrane conductances in epidermal cell pairs isolated from the prepupal integument of the flour beetle, Tenebrio molitor. The mean initial junctional conductance in 267 cell pairs was 9.5 +/- 1.0 nS (range 0-95 nS). Well-coupled cell pairs uncoupled spontaneously with a half-time of 7.6 min. Adding 5 mM ATP to the pipette solution stabilized coupling with less than a 50% drop occurring after 30 min. Nonjunctional membrane potential was the major determinant of junctional conductance with transjunctional potential playing a minor role. Junctional conductance approached 0 pA at nonjunctional membrane potentials greater than 0 mV and increased with hyperpolarization. The voltage at half maximal conductance was -26 mV. The time course of the reversible changes in junctional conductance were slow (< or = 30 sec) with time-dependent decay occurring faster and recovery occurring slower with increasing depolarization. Single gap junctional channel activity was recorded in uncoupling cell pairs and in poorly coupled ATP-stabilized cell pairs. One main single channel conductance was observed in each cell pair. The mean single channel conductances from all cell pairs in this study ranged from 197-347 pS (mean 248 pS). Single channel conductance was linear over the +/- 60 mV transjunctional voltage range tested. A broad range of subconductance states of the main state representing 5% of the total open time of measurable main state events was observed. Single channel activity was strongly dependent on the nonjunctional membrane potential, increasing with hyperpolarization. PMID- 8411138 TI - Structure/function studies of the epithelial isoforms of the mammalian Na+/H+ exchanger gene family. PMID- 8411139 TI - Viruses accumulate spontaneously near droplet surfaces: a method to concentrate viruses for electron microscopy. AB - Virus particles suspended in a drop of water tend to concentrate at or near its surface with the air. The concentrated, and probably more purified, particles may then be collected on a film-coated grid for negative staining and electron microscopy. This is a useful method, simpler than others (e.g. high-speed centrifugation, Lyphogel, or precipitation by (NH4)2SO4) which are used to process clinical specimens for diagnosis where virus particles may be too dilute in the original sample. It is shown, by freeze-fracturing for electron microscopy, that Orf virus particles do accumulate at and just below the surfaces of drops. Various physical effects which may cause the particles to accumulate are considered. Results from a computer model suggest that Brownian motion alone could be adequate to transport a useful quantity of the particles in the body of a 2-mm-diameter hemispherical drop to its surface if the particles do not clump and if they remain trapped at the surface when they reach it. In practice, transport by Brownian motion is likely to be augmented by swirling, convection and other effects within drops. PMID- 8411140 TI - Second-order stereology of benign and malignant alterations of the human mammary gland. AB - The purpose of the present study was a quantitative characterization of the three dimensional arrangement of the epithelial component of benign and malignant alterations of the female breast by combining stereology with stochastic geometry. Twenty cases of fibrous mastopathy and 20 cases of invasive ductal mammary cancer were studied at the light microscopic level. Segmentation of the epithelial tissue component was performed with an image analyser. From the resulting binary images, unbiased estimates of the covariance C(r) and the intensity Vv of the epithelial volume component were obtained automatically by computer. From these data, estimates of the correlation function k(r), of the pair correlation function g(r), of the radial distribution function RDF(r) and of the reduced second moment function K(r) of epithelial volume were determined. The estimates of C(r) and RDF(r) differed between groups, but these functions depend on spatial pattern and Vv. As carcinomas showed a significantly higher epithelial volume density Vv than mastopathies, estimation of C(r) and RDF(r) alone did not permit a safe distinction between possibly different types of spatial arrangement of epithelium in the benign and malignant lesions. Analysis of the estimates of k(r), g(r) and K(r), which are not influenced by Vv, showed definite interaction between epithelial volume elements, with clustering at short distances and repulsion at long distances. In both groups, the null hypothesis of purely random arrangement of epithelium had to be rejected. The clearest distinction between groups was obtained by estimation of g(r), which showed that short-range, tubular pattern as well as long-range, lobular architecture are better preserved in benign than in malignant lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411141 TI - Social relationships and health-related behavior. AB - The relationship of social ties to health behavior is studied in this paper. The general hypothesis tested is that those with social relationships are more likely to avoid health damaging behavior. Patterns regarding specific social ties and particular health behaviors are examined. Findings indicate the relationships of spouse, organization member, and friend are related to preventive health behavior; spouses are less likely smokers, drinkers, and heavy drinkers. The friend relationship is related to decreased smoking behavior, while the employee relationship is related to increased drinking. A consistent predictor of health behaviors is the social tie of organization member. People with relationships as organization members are more likely always to wear seat belts, smoke and drink less, and consume lower amounts of these substances. PMID- 8411142 TI - The family's functioning with newly diagnosed breast cancer in the mother: the development of an explanatory model. AB - Despite the high rates of breast cancer in the child-rearing mother, there is extremely limited research on the effects of the illness on the children, marriage, and parent-child relationship. The current study tested an explanatory model of family functioning with breast cancer based on data obtained from standardized questionnaires from 80 diagnosed mothers and partners with young school-age children. Path analysis results for data obtained from both the mothers and the partners revealed a similar pattern. More frequently experienced illness demands were associated with higher levels of parental depressed mood which negatively affected the marriage. When the marriage was less well adjusted, it negatively affected the family's coping behavior. Household functioning was positively affected by heightened coping activity and by higher levels of marital adjustment. Children functioned better when the non-ill parent more frequently interacted with them and their families coped more frequently with their problems. PMID- 8411143 TI - Component analysis of the Structured Interview for assessment of Type A behavior in employed women. AB - Components of the Structured Interview in employed women and the overlap between these components and those identified by self-report measures of Type A behavior were investigated. The sample consisted of 177 employed, white females 26 to 52 years old; 48% were classified as Type A. Principal components factor analysis yielded five factors: Clinical Rating, Hurried-Drive, Impatience, Pressured Competitiveness, and Expression of Anger. Some differences in the factor structure were noted for this sample of employed women, compared with previous studies of employed men, particularly with respect to anger and impatience. The behavioral components of the Structured Interview did not correlate with self report measures of Type A behavior (Framingham Type A scale and the Jenkins Activity Scale). Implications for future studies of employed women are discussed. PMID- 8411144 TI - Menopausal symptoms as consequences of dysrhythmia. AB - A theoretical model is presented for understanding physical and psychological changes which frequently accompany the menopausal transition. According to this, as yet untested, model, as the menopausal transition progresses, hormonal patterns become unpredictable and previously learned homeostatic mechanisms can no longer provide a balanced internal milieu. The consequences of this dysrhythmia are the physical and psychological disruptions typical of jet travel and shift work. Internal attributions of illness and deficiency, promoted by the medical model and the popular media, have transformed these disruptions into a universal "deficiency disease." The model represents a theoretical integration and synthesis of diverse and, historically, unrelated areas; it is presented in order to promote empirical research on the causes, correlates, and interventions of menopausal symptoms. PMID- 8411145 TI - Psychological stress reverses antiaggregatory effects of dietary fish oil. AB - Effect of acute psychological stress on the inhibition of in vitro platelet aggregation by dietary long-chain n-3 fatty acids was studied in 20 adult males. Subjects were randomly divided into groups receiving either olive oil or fish oil (2.4 g long-chain n-3 fatty acids/day) for 4 weeks. In vitro aggregation responses to two doses of ADP collagen, and epinephrine were measured immediately prior to and following exposure to three psychological stressors (2 min each), before and after the supplementation period. Olive oil had no effect on baseline aggregatory responses, while fish oil reduced aggregatory responses to ADP and epinephrine. Exposure to the stressors had no effect upon presupplementation aggregation in either group or in the olive oil group postsupplementation. However, stress abolished antiaggregatory effects of fish oil. This reversal of the antiaggregatory effects of fish oil by mild stress suggests possible limitations of low-dose fish oil supplementation in clinical situations. PMID- 8411146 TI - The favorite cigarette of the day. AB - The choice of a favorite cigarette smoked during a given day varies considerably. Preferences are related to nicotine dependence and the lifestyles of smokers. In a sample of 5124 smokers, the "after-dinner cigarette" was chosen as the cigarette which they would miss most, while the next largest proportion of smokers (33%) said that they would miss the "first cigarette of the morning" most. The latter group scored highest on tests of nicotine dependence. In contrast, infrequent smokers ("chippers") chose the after-dinner cigarette. PMID- 8411147 TI - Identification of the contact surface of a streptococcal protein G domain complexed with a human Fc fragment. AB - The B1 domain of streptococcal protein G interacts with the C-terminal fragment of the heavy chain of immunoglobulin G (IgGFc). The binding site for the protein G domain on the antibody fragment is in close proximity or overlapping with that determined for staphylococcal protein A. The interaction of the B1 domain with IgGFc was investigated by 1H-15N correlation spectroscopy. The major interaction site on the B1 domain comprises parts of beta-strand 3 as well as the alpha helix. Comparison with the crystal structure of the protein A/IgGFc complex suggests that the mode of interaction in the two complexes is analogous, despite the lack of sequence or structural similarity between two antibody binding proteins. PMID- 8411148 TI - Interactions of NodD at the nod Box: NodD binds to two distinct sites on the same face of the helix and induces a bend in the DNA. AB - The Rhizobium meliloti nodD gene products are positive transcriptional activators of genes required for early stages of nodule morphogenesis in the R. meliloti alfalfa symbiosis (nod genes). The regulatory activity of NodD, a member of the LysR family of activator proteins, is mediated in part through its binding to conserved DNA sequences termed nod boxes which lie upstream of the inducible nod genes. Here we use interference footprinting to identify two NodD binding sites in the nodA, nodF and nodH nod boxes. These two binding sites are located on the same face of the DNA helix and can be separated by an additional 10 bp with retention of activity. By systematic alteration of the phasing of the two binding sites on the DNA helix, we showed that only constructs which contain both sites on the same side of the helix are recognized by NodD as determined by migration retardation assay and by in vivo activation of nod box-lacZ fusions. Moreover, NodD apparently induces a bend in the DNA upon binding at the nod box as shown by migration retardation behavior of circularly permuted nod box fragments. PMID- 8411149 TI - A new start site for Escherichia coli RNA polymerase at an engineered short region of non-complementarity in double-stranded DNA. AB - We have constructed versions of the bacteriophage PRM promoter containing short (9 or 12 base pairs) regions of DNA mismatches ("bubble") which include the authentic transcription start site of the unmodified promoter. These constructs direct transcription initiation at positions near the genuine PRM start site. In addition a new start site (designated Pbub) is observed in the region of non complementarity, from which RNA synthesis proceeds in the opposite direction. The ability to initiate the divergent transcripts is specific to holo enzyme. Mapping of the Pbub start sites shows that they are but a few base pairs upstream of the edge of the bubble. Thus, with respect to the single-stranded region, the location of the start site is no different for Pbub than it is for open complexes at promoters. Compared with an unmodified PRM promoter, the region protected by RNA polymerase from digestion by DNase I is extended in the downstream direction (with respect to the PRM start) at the promoters bearing mismatches; this is consistent with the binding of the divergently transcribing RNA polymerase. Interestingly, cI protein represses rather than activates RNA synthesis originating in the PRM direction, indicating yet another aspect in which the complexes formed at these constructs differ from open complexes at the unmodified promoter. PMID- 8411150 TI - Yeast a1 and alpha 2 homeodomain proteins form a DNA-binding activity with properties distinct from those of either protein. AB - The yeast a1 and alpha 2 proteins are examples of homeodomain proteins that display cell-type-specific expression. They are co-expressed in only one type of cell, the a/alpha cell, where they repress the expression of a group of target genes. Using purified proteins, we demonstrate that a1 and alpha 2 form a highly specific DNA-binding activity, which recognizes an operator found upstream of each target gene. These proteins interact with DNA to form a ternary complex in which both a1 and alpha 2 contact the DNA through their respective homeodomains. An alpha 2 homodimer can recognize the same operator as the a1/alpha 2 heterodimer, but the affinity and specificity of the alpha 2 homodimer for DNA are much weaker than those of the a1/alpha 2 heterodimer. This difference results in part from the fact that the heterodimer is formed using a set of protein protein contacts that is distinct from those that form the alpha 2/alpha 2 homodimer. Although a1 contacts DNA in the presence of alpha 2, it does not on its own bind DNA specifically, even at very high concentrations. These results suggest that the dimerization of heterologous partners can produce a DNA-binding activity that is not a simple hybrid of the known properties of each constituent. PMID- 8411151 TI - The gene clusters ARC and COR on chromosomes 5 and 10, respectively, of Saccharomyces cerevisiae share a common ancestry. AB - The yeast Saccharomyces cerevisiae contains two clusters of eight genes each on chromosome 10 and 5, denoted, respectively, the COR and ARC regions. The genes in the COR region include TRS1 (a tRNA(Ser) gene), ANB1, CYC1, UTR1, UTR3, OSM1, tRNA(Gly) and RAD7 whereas the genes in the ARC region include TRS2 (a tRNA(Ser) gene), TIF51A, UTR5, ANP1, RAD23, UTR4, CYC7 and UTR2. We have performed a physical analysis of the ARC region, including determining DNA sequence of the 7529 nucleotides; the open reading frames; the size and orientation of the transcripts; and the phenotypes resulting from deletions or gene disruptions. The ARC region was systematically compared to the COR region which was previously described. The gene pairs CYC1-CYC7 and ANB1-TIF51A were previously shown to be, respectively, approximately 80% and 90% identical. tRNA(Ser) genes, TRS1 and TRS2, are located in both clusters 953 nt and 344 nt downstream of ANB1 and TIF51A, respectively. Some of the other gene pairs of these clusters are related in function and share only short segments of similarity distributed within the regions. The best alignment based on amino acid and nucleotide sequences indicates that the ARC and COR regions are ancestrally related by a duplication, a transposition, and a single rearrangement, followed by extensive divergence. These comparisons allowed an evaluation of distantly related sequences not obviously revealed by standard computer analysis. Surprisingly, the alignment suggested that a translated region of the ARC ANP1 gene and the COR tRNA(Gly) gene are ancestrally related. Also translated regions of the COR gene RAD7 share similarities with both of the two adjacent ARC genes, ANP1 and RAD23. Five examples of simple repeated amino acid and DNA sequences occurred in the ARC region but none in the COR region. We suggest that these repeated sequences played a role in the divergence of ARC genes. PMID- 8411152 TI - Origin of the asymmetrical contact between lac repressor and lac operator DNA. AB - The Escherichia coli lac operator DNA contains two sequence repeats related by a pseudo-dyad axis. Deviations from symmetry, in the central 21 bp sequence, occur at two pairs of symmetrically related sites (+15/+7, +13/+9) and at the central base-pair +11. Mutational analysis and DNA protection studies have suggested asymmetric interactions of lac repressor along this sequence. Previous biophysical studies on the lac repressor-operator system have typically employed symmetrized operator sequences to simplify analysis. As a result, it has remained difficult to assess the importance of the naturally occurring sequence deviations from symmetry. Here, 19F-NMR is used to determine if the wild-type E. coli lac operator DNA sequence itself specifies a pair of distinct half-site interactions with lac repressor DNA binding domains. To observe protein interactions simultaneously at operator half-sites using 19F-NMR, three pairs of naturally occurring, symmetry related thymine residues (at +6/+16, +8/+14 and +1/+21) were substituted pair-wise by 5-fluorodeoxyuridines (5-FdU). Two polypeptides corresponding to the N-terminal DNA binding domain of lac repressor "headpiece", residues 1 to 56 and 1 to 64, were employed to remove the steric constraints of subunit interaction in the wild-type tetramer. Spectral changes associated with headpiece binding to left side DNA sequences differ from those caused by binding to equivalent sequences on the right half-site. These results are similar to non symmetric intact tetramer repressor interactions specified by the DNA sequence. Three mutant lac operator sequences with increased symmetry, bearing FdU substitutions were used to identify the relative importance of the three naturally asymmetric positions. Symmetrizing one pair of these sites alone or in addition to removing the central base-pair failed to produce identical NMR signal changes characteristic of symmetric headpiece-DNA complexes. However, symmetrizing both asymmetric pairs gave chemical shift changes expected from symmetric protein-DNA complexes. We propose that key interactions with the left side +9 (G.C) are altered at the symmetrically related right side +13 (A.T). The data show that the DNA sequence at +13 influences interactions three base-pairs away. PMID- 8411153 TI - Plasmid addiction genes of bacteriophage P1: doc, which causes cell death on curing of prophage, and phd, which prevents host death when prophage is retained. AB - P1 lysogens of Escherichia coli carry the prophage as a stable low copy number plasmid. The frequency with which viable cells cured of prophage are produced is about 10(-5) per cell per generation. Here we show that a significant part of this remarkable stability can be attributed to a plasmid-encoded mechanism that causes death of cells that have lost P1. In other words, the lysogenic cells appear to be addicted to the presence of the prophage. The plasmid withdrawal response depends on a gene named doc (death on curing), encoding a 126 amino acid protein. Expression of doc is not SOS-inducing and killing by Doc is recA independent. In cells that retain P1 the killing is prevented by the product of a gene named phd (prevent host death), encoding a 73 amino acid protein. The genes phd and doc have been cloned and expressed from a 0.7 kb segment of P1 DNA. The two genes constitute an operon and the synthesis of Doc appears to be translationally coupled to that of Phd. Homologs of the P1 addiction genes are found elsewhere, but phd and doc are unrelated to previously described genes of other plasmids that also cause an apparent increase in plasmid stability by post segregational killing. PMID- 8411154 TI - Dual role of the sequence-specific bacteriophage T4 endoribonuclease RegB. mRNA inactivation and mRNA destabilization. AB - Gene regB of bacteriophage T4 encodes a sequence-specific endoribonuclease that introduces cuts in early phage messenger RNAs. Cutting takes place specifically in the middle of the tetranucleotide GGAG, as soon as the first minute of infection. Out of the 20 processing sites so far identified, seven are in Shine Dalgarno sequences. The others are localized in intercistronic regions or within coding sequences. In the latter case, cutting efficiency is much lower. regB dependent cleavages can occur within AU-rich sequences downstream of processed GGAG motifs that are not in effective translation initiation sites. We looked for possible consequences of regB-dependent cuts on gene expression in two early regions of the T4 chromosome. In the comC alpha region, none of the three major RegB cleavage sites is in a Shine-Dalgarno sequence, and in the motA region the unique regB-dependent processing site is found within the Shine-Dalgarno sequence of the gene. We find that in the region of gene comC alpha, RegB decreases two- to threefold the chemical half-life of early transcripts, but does not change the functional half-life of mRNAs coding for protein ComC alpha. The amount of MotA protein synthesized by the wild-type is half that obtained in a regB mutant infection. We show that this is a direct consequence of mRNA processing by RegB at the Shine-Dalgarno sequence of motA. This regB-mediated translation inhibition is not accompanied by an important modification in motA mRNA chemical half-life. We show that rapid shut-off of MotA protein synthesis that occurs soon after infection results both from RegB processing within the translation initiation region of motA and from early transcription inhibition followed by regB independent breakdown of the motA mRNA. PMID- 8411155 TI - fd coat protein structure in membrane environments. AB - The membrane bound form of bacteriophage fd coat protein has a long hydrophobic membrane spanning helix and a shorter amphipathic helix in the plane of the bilayer. Residues near the N and C termini and in the turn connecting the two helices are mobile. The locations and orientations of the helical secondary structure elements and the protein backbone dynamics were characterized by combining results from multidimensional solution NMR experiments on protein samples in micelles and high resolution solid-state NMR experiments on protein samples in oriented and unoriented lipid bilayers. The coat protein is a monomer in micelles. The secondary structure of the membrane bound form of fd coat protein is very similar to that of the structural form found in the virus particles, since it is nearly all alpha helix. However, the membrane bound form of the protein differs from the structural form of the protein in virus particles in the arrangement of the secondary structure, since the membrane bound form of the protein has two distinct helical domains oriented perpendicular to each other and the structural form of the protein in the virus particles has a nearly continuous helix aligned approximately along the filament axis. In addition, there are substantial differences in the dynamics of residues in the bend between the two helices and near the C terminus, since they are mobile in the membrane bound form of the protein and not in the virus particles. Residues 1 to 5 at the N terminus are highly mobile and unstructured in both the membrane bound and structural forms of the coat protein. PMID- 8411156 TI - Packing of secondary structural elements in proteins. Analysis and prediction of inter-helix distances. AB - Interacting helix pairs were defined as those in which each member has one residue which undergoes a fractional loss of 0.10 solvent accessible contact area on association. Analysis of 1095 such pairs of helices, selected from the Brookhaven Data Bank, confirms that helix pairs in proteins can pack interactively at all angles with preferential packing observed in the angle intervals (omega), -160 degrees to -140 degrees, -100 degrees to -20 degrees, 10 degrees to 40 degrees, and 50 degrees to 160 degrees. The distance (d) between the helix axes in the contact region of two interactively packed helices is linearly correlated with the volume-dependent function, log(V/nda), of the residues in this region. The correlation is used to predict inter-helix distances in equivalent helix pairs of homologous proteins and is therefore of value in comparative modelling of protein three-dimensional structures. PMID- 8411157 TI - A local alignment method for protein structure motifs. AB - A method for the comparison of protein three-dimensional substructures was developed. The method employs the double dynamic programming method of Taylor & Orengo but identifies multiple local alignments rather than a single global alignment. A modification based on the Smith Waterman algorithm for sequence alignment enables the automatic identification and growth of the most structurally similar local alignments irrespective of length and composition. The method can also be used in a search mode to match substructures. Roughly five minutes is required to find all Greek-key motifs in a protein of 100 residues. Two novel globin folds have been detected using the method, the antibiotic protein colicin A and diphtheria toxin, neither of which have any functional relationship to the globins. PMID- 8411158 TI - X-ray crystal structure of ferric Aplysia limacina myoglobin in different liganded states. AB - The X-ray crystal structure of the ligand-free ferric form of Aplysia limacina myoglobin (pH 6.0) has been refined at 1.7 A resolution (R = 15.1%), and its cyanide, thiocyanate and imidazole derivatives studied by difference Fourier techniques at atomic resolution. The crystallographic R-factors of the three different derivatives reported are 16.1%, 16.1% and 15.6% at 1.8 A, 2.0 A and 2.0 A resolution, respectively. The present results have been analyzed in parallel with previous crystallographic studies on the molecular structures of the fluoride and azide derivatives of ferric Aplysia limacina myoglobin. Ligand binding to the distal site of the heme pocket results in different networks of hydrogen bonds involving to various degrees the bound ligand, residue Arg(66)E10, the heme propionate III, ordered water molecules and/or protein backbone atoms from the CD region. In particular, Arg(66)E10 stabilizes the bound ligand and compensates for the absence of the hydrogen bond donor residue HisE7, commonly present in oxygen-carrying globins. PMID- 8411159 TI - Structure of a DNA:RNA hybrid duplex. Why RNase H does not cleave pure RNA. AB - The solution structure of the DNA:RNA hybrid duplex d(GTCACATG):r(caugugac) has been determined by means of two-dimensional nuclear Overhauser effect (2D-NOE) spectra, restrained molecular dynamics and full-relaxation matrix stimulation of the 2D-NOE spectra. The DNA:RNA hybrid duplex assumes neither an A-form nor a B form structure in solution, but an intermediate heteromerous duplex structure. The sugars of the RNA strand have a normal N-type C3'-endo conformation, but the DNA strand sugars have neither N-type nor S-type conformations; instead, they have an unexpected intermediate O4'-endo conformation. The negative x displacement, as well as the small rise and positive inclination of the base pairs, resembles A-form morphology but the minor groove width is intermediate between that of A-form and B-form duplexes. Both the DNA and RNA strands show prominent sequence-dependent variations in their helical parameters. Combined analysis of NOE and J-coupling data indicates that the DNA sugars are not in a dynamical two-state equilibrium. The detailed three-dimensional structure of this DNA:RNA hybrid molecule leads to a proposed model for its interaction with RNase H. Several specific structural features of the enzyme complexed with the hybrid duplex appear to explain the mechanism whereby RNase H discriminates between DNA:RNA hybrid duplexes and pure RNA:RNA duplexes. PMID- 8411160 TI - A novel allosteric mechanism in haemoglobin. Structure of bovine deoxyhaemoglobin, absence of specific chloride-binding sites and origin of the chloride-linked Bohr effect in bovine and human haemoglobin. AB - The structure of bovine deoxyhaemoglobin has been determined at 2.2 A resolution and refined to an R-factor of 0.193 for all 32,583 reflections, and a free R factor of 0.249 for 1527 reflections excluded from the refinement. The structure shows no significant differences between the alpha-carbon positions of bovine and human haemoglobin, except at the N-terminal segment and the first helix (A) which are closer to the dyad symmetry axis and pushed more tightly against the rest of the beta-subunits in the bovine form. In a search for the predicted chloride binding sites, three-dimensional data were collected from crystals suspended in 50% polyethylene glycol buffered either with 50 mM Na phosphate (pH 7.3) +/- 0.1 M NaCl or with 0.1 M Hepes (pH 7.3) +/- 0.1 M NaBr. Difference electron density maps with and without NaCl or NaBr showed no evidence of specific halide ion binding sites. Oxygen equilibria were measured in 10 mM Hepes buffer without added NaCl, with 0.1 mM NaCl, 0.1 M NaCl + 1 mM 2,3-diphosphoglycerate, and 0.1 M NaCl + 1 mM inositol hexaphosphate. Without added chloride, P50 of stripped bovine haemoglobin was similar to that of human haemoglobin with 0.1 M NaCl. With 0.1 M NaCl it was similar to that of human haemoglobin saturated with 2,3 diphosphoglycerate. In 0.1 M NaCl neither organic phosphate significantly affected the oxygen affinity. Titration of P50 with NaCl showed delta log P50/delta log[Cl-] of bovine and human haemoglobin to be identical. Analysis of the oxygen equilibrium curves showed the low intrinsic oxygen affinity of bovine haemoglobin to be due to a larger oxygen dissociation constant from the T structure. The influence of chloride on P50 and on the alkaline Bohr effect is the same in bovine and human haemoglobins. It is proposed that this is due to the excess positive charges in the central cavity and its widening in the transition from the R to the T-structure. The widening would allow more chloride ions to enter and neutralize the positive charges, but these ions would remain mobile and therefore do not show up as peaks of high electron density. Repulsion between excess positive charges in the central cavity raises the free energy of the T structure relative to the R-structure, thereby raising the oxygen affinity. Conversely, entry of chloride ions on widening of the cavity reduces the free energy of the T-structure and therefore lowers the oxygen affinity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411161 TI - Crystallization and preliminary X-ray studies of human erythrocyte acylpeptide hydrolase. AB - Crystals of acylpeptide hydrolase suitable for structure determination have been obtained. This enzyme removes the N-terminal formyl or acetyl group together with the first amino acid residue from N-terminal blocked peptides including bioactive peptides. One set of crystals, which diffract to 2.2 A, are in space group P2 with cell dimensions a = 118.6 A, b = 82.3 A, c = 182.1 A, beta = 91.6 degrees. The search for suitable heavy-atom derivatives is underway. PMID- 8411162 TI - Crystallization and preliminary X-ray studies of extracellular signal-regulated kinase-2/MAP kinase with an incorporated His-tag. AB - The extracellular signal-regulated kinase ERK2, a member of the protein kinase superfamily, phosphorylates a variety of cellular proteins in response to extracellular signals. ERK2 expressed in Escherichia coli as a fusion protein with the sequence Ala-His6 at the N terminus has low basal activity and very low levels of phosphate incorporation, but can be fully activated. The Ala-His6 ERK2 as expressed in the unphosphorylated form has been crystallized in space group P2(1). The cell constants are a = 49.32 A, b = 71.42 A, c = 61.25 A, and beta = 109.75 degrees, and the crystals diffract to better than 1.8 A resolution. PMID- 8411163 TI - Crystallization and preliminary X-ray diffraction analysis of the human dimeric S Lac lectin (L-14-II). AB - The human recombinant S-Lac lectin, L-14-II, produced in an Escherichia coli expression system, has been co-crystallized in the presence of lactose by the hanging drop vapor diffusion method. The crystals grow in space group P2(1)2(1)2(1) with unit cell dimensions of a = 43.6 A, b = 57.8 A, c = 108.2 A, with a dimer in the asymmetric unit. On a conventional rotating anode the crystals diffract to at least 2.8 A resolution. PMID- 8411164 TI - The redox properties of protein disulfide isomerase (DsbA) of Escherichia coli result from a tense conformation of its oxidized form. AB - Periplasmic protein disulfide isomerase (DsbA) from Escherichia coli is a strongly oxidizing thiol reagent with one catalytic disulfide bridge and an intrinsic redox potential of -0.089 V. Gel filtration experiments and analytical ultracentrifugation studies demonstrate that DsbA is a monomeric protein with a molecular mass of 21.1 kDa, independent of its redox state. In order to investigate the molecular basis of its redox properties, the guanidinium.chloride induced folding/unfolding equilibrium of the reduced and the oxidized form of the enzyme were compared. The transitions at pH 7.0 and 30 degrees C were found to be fully reversible and allowed the calculation of the free energy of stabilization of oxidized and reduced DsbA according to a two-state model for the unfolding transition. The analysis reveals that reduced DsbA is 22.7 (+/- 4.0) kJ/mol more stable than oxidized DsbA. This energetic difference is essentially independent of temperature, although the overall free energies of stabilization of both oxidized and reduced DsbA vary strongly between 20 and 30 degrees C as a consequence of changes in the cooperativity of the transitions The conformational tension of 22.7 (+/- 4.0) kJ/mol in oxidized DsbA quantitatively explains the oxidizing properties of the protein, as it causes a change of redox equilibrium constants between DsbA and thiols of about four orders of magnitude, corresponding to an increase of the standard redox potential of 0.118 (+/- 0.021) V. We conclude that the oxidizing properties of DsbA mainly result from a tense conformation of its oxidized form, that is converted to the relaxed, reduced state upon oxidation of thiols by DsbA. The results are discussed in terms of a general principle underlying the oxidizing properties of protein disulfide isomerases. PMID- 8411165 TI - The function of individual zinc fingers in sequence-specific DNA recognition by transcription factor IIIA. AB - Mutations in Zn(2+)-coordinating histidine residues have been used to generate forms of transcription factor IIIA containing structural disruptions in single zinc fingers. These mutant proteins have been analyzed with respect to the structural and functional independence of individual zinc fingers in TFIIIA. They have also been used to assess the energetic contributions to binding and the sites of interaction of each of the nine zinc fingers of TFIIIA with the 5 S rRNA gene. The results are surprising and suggest a complex mode of binding in which the interactions of structurally independent zinc fingers with the DNA substrate are non-uniform and functionally interdependent in a way that may help to explain some of the unusual properties of TFIIIA. PMID- 8411166 TI - Identification of the auxin-responsive element, AuxRE, in the primary indoleacetic acid-inducible gene, PS-IAA4/5, of pea (Pisum sativum). AB - The plant hormone auxin transcriptionally activates early genes in pea. We have developed a transient assay system using protoplasts of auxin-responsive pea seedling cells to define the auxin-responsive element, AuxRE, of the early auxin induced PS-IAA4/5 gene. The auxin responsive protoplasts show an authentic hormonal response identical to that observed in intact pea tissue, with respect to rapidity, specificity and cycloheximide (CHX) inducibility of the PS-IAA4/5 transcript. The hormone also mediates rapid and specific induction of chloramphenicol acetyltransferase (CAT) activity in protoplasts transfected with a chimeric IAA4/5-CAT gene. The IAA-induced CAT activity is developmentally regulated and is observed only in protoplasts derived from auxin-responsive regions of the pea seedling. Extensive deletion analysis of the PS-IAA4/5 promoter defined a promoter region between -318 and -154 that confers auxin inducibility. This AuxRE mediates auxin-inducible CAT activity in pea cells driven by the non auxin-responsive CaMV 35S minimal promoter. The functionality of this promoter region as an AuxRE was further verified in tobacco plants using IAA4/5-GUS gene fusions. The AuxRE contains two domains: Domain A acts as an auxin switch; domain B has an enhancer-like activity. The A and B domains contain the highly conserved sequences found in various auxin-regulated genes (T/GGT CCCAT (domain A) and C/AACATGGNC/AA/GTGTT/CT/CC/A (domain B)). DNase I footprinting reveals binding of nuclear proteins to the highly conserved sequence found in A and B domains. The sequence of the A domain does not correspond to any known regulatory elements found in other eukaryotic genes, and the data suggest that this conserved motif functions as an AuxRE. A model for the early transcriptional activation of the PS-IAA4/5 gene by IAA is discussed. PMID- 8411167 TI - Two inactive fragments derived from the yeast mitochondrial ribosomal protein MrpS28 function in trans to support ribosome assembly and respiratory growth. AB - The mitochondrial ribosomal protein MrpS28 of Saccharomyces cerevisiae is one of several mitochondrial ribosomal proteins homologous to Escherichia coli ribosomal proteins within the context of a larger protein. Relative to a region of homology with E. coli ribosomal protein S15, the mature MrpS28 protein has unique sequence domains of 117 and 48 amino acids at its amino and carboxyl terminus, respectively. To better understand the role of the various sequence domains of the MrpS28 protein in vivo, truncated derivatives were expressed under conditions where they were the only potential source of functional MrpS28 protein. The results shown here demonstrate that the amino-terminal domain and the S15-like domain are both essential for respiratory growth. Interestingly an inactive amino terminal fragment can be complemented in trans by a second inactive fragment comprising the S15-like domain and the carboxyl-terminal 48 amino acids. Consequently, the assembly of these fragments into ribosomal subunits can be examined when they are expressed individually or together. Results from these studies indicate that each of the MrpS28-derived fragments facilitates the incorporation of the other into 37 S ribosomal subunits. PMID- 8411168 TI - Derivatives of the yeast mitochondrial ribosomal protein MrpS28 replace ribosomal protein S15 as functional components of the Escherichia coli ribosome. AB - The mitochondrial ribosomal protein MrpS28 is considerably larger than its eubacterial homolog, Escherichia coli ribosomal protein S15 (Eco S15). Relative to a region of homology that spans the entire length of the bacterial protein, mature MrpS28 is extended by 117 and 48 amino acids at its amino and carboxyl termini, respectively. Both the amino-terminal and S15-like domains of MrpS28 are essential for function in yeast mitochondria. Here, we show that these same two domains function in E. coli. The S15-like domain of MrpS28 alone complements a cold-sensitive mutation in E. coli strain KR121 that gives rise to reduced levels of Eco S15. However, complementation by the S15-like domain of MrpS28 is inefficient when compared with Eco S15. Surprisingly, the amino-terminal domain of MrpS28, which is apparently a unique component of the mitochondrial ribosome and is unable by itself to complement the cold-sensitive phenotype, enhances the ability of the S15-like domain to support growth of KR121 cells at nonpermissive temperatures. Together, these data suggest that the amino-terminal domain contributes to the fundamental properties of MrpS28 involved in the assembly and function of both mitochondrial and E. coli ribosomes. PMID- 8411169 TI - Two acidic residues of Escherichia coli methionyl-tRNA synthetase act as negative discriminants towards the binding of non-cognate tRNA anticodons. AB - Escherichia coli methionyl-tRNA synthetase recognizes its cognate tRNAs according to the sequence of the CAU anticodon. In order to identify residues of methionyl tRNA synthetase involved in tRNA anticodon recognition, enzyme variants created by cassette mutagenesis were genetically screened for their acquired ability to charge tRNA(mMet) derivatives with an ochre or an amber anticodon and, consequently, to cause the suppression of a stop codon in an indicator gene. The selected enzymes are called suppressors. Mutations were firstly directed towards the region of the synthetase encompassing residues 451 to 467. Several dozens of suppressor enzymes were compared. Statistical analysis of the mutations suggested that the substitution of an Asp side-chain at position 456 was sufficient to render possible the charging of the ochre or amber suppressor tRNAs. Point mutants at this position were therefore constructed. Their behaviour demonstrated that various tRNA(Met) derivatives having a non-Met anticodon could be aminoacylated in vitro provided only that the side-chain of residue 456 was no longer acidic. In turn, the Asp456 residue is not essential to the CAU anticodon recognition, since its substitution does not impair the aminoacylation of wild type tRNA(Met). The analysis was enlarged to a second region from residue 437 to residue 454. The mutagenesis highlighted two other positions, one of which, Asn452, appeared involved in wild-type tRNA(Met) binding. The second position, Asp449, plays a role very similar to that of Asp456. It is concluded that both Asp449 and 456 behave as "antideterminants", contributing together to the rejection by the enzyme of tRNAs carrying non-Met anticodons. Finally, it is shown that the activities of some particular methionyl-tRNA synthetase variants, which have been made indifferent to the sequence of the anticodon of a tRNA(Met), are tightly dependent on the presence of the nucleotide determinants specific to the acceptor stem of tRNA(Met). PMID- 8411170 TI - A shortened form of the Tetrahymena thermophila group I intron can catalyze the complete splicing reaction in trans. AB - The group I intron from Tetrahymena thermophila is able to catalyze its own excision from a precursor RNA. The intron recognizes the splice sites through an intron-encoded sequence called the internal guide sequence, or IGS. The 5' and 3' exons are thought to align on the IGS and form a pseudoknot structure consisting of two stems (P1 and P10). We created a shortened form of the intron that lacks the exon sequences and the entire IGS. This RNA is unable to react upon itself. It can catalyze a sequential two-step transesterification reaction on a P1P10 substrate added in trans that completely mimics splicing. The reaction works for different substrates that contain a U.G base-pair preceding the 5' cleavage site and a guanosine base preceding the 3' cleavage site, but that are otherwise unrelated in sequence. The ribozyme uses primarily the correct 5' and 3' splice sites even in the presence of potential cryptic splice sites, and therefore it must rely on the structure of the substrate (formation of the P1 and P10 helices) for correct splice site recognition. A C-G base-pair after the 5' splice site in P1 decreases activity while a U.G or G.U base-pair enhances activity. The relative position in P1 of the U.G base-pair preceding the 5' splice site is an important determinant. The ability of the intron to recognize primarily a specific structure, rather than a sequence, has ramifications for splice-site selection, for molecular modeling of the group I intron, and for ribozyme-based gene targeting. PMID- 8411171 TI - Basal promoter and enhancer element of yeast U6 snRNA gene. AB - The yeast U6 snRNA gene, SNR6, transcribed by RNA polymerase III or C, is shown to have a mixed promoter with upstream, intragenic and downstream elements. The distant downstream B block behaves as a typical enhancer element. Required in vivo, and for transcription of chromatin templates in vitro, it was also active in reversed orientation. As shown by footprinting and electron microscopy, the factor TFIIIC, or tau, bound the B block in an oriented manner and was able to induce DNA looping. The factor TFIIIC appeared to act via a weak A block located at position +21. This A block-related motif was essential in vivo and with chromatin templates. When changed into a consensus A block it favored DNA looping by TFIIIC firmly anchored on the B block, and activated a B block lacking gene in vivo and in vitro. The role of the TATA box at -30 was most apparent using a purified transcription system. With the A block, it appeared to contribute to start site selection. The results suggest a model where three weak promoter elements collaborate to assemble the transcription complex by DNA looping and synergistic protein-DNA interactions. PMID- 8411172 TI - Genetic analysis of periplasmic binding protein dependent transport in Escherichia coli. Each lobe of maltose-binding protein interacts with a different subunit of the MalFGK2 membrane transport complex. AB - Escherichia coli is able to accumulate maltose and maltodextrins by an ATP binding cassette transporter known as the maltose transport system. This transport system is comprised of five proteins: the LamB protein in the outer membrane; the periplasmic maltose-binding protein (MBP); two integral inner membrane proteins, MalF and MalG; and MalK, which is associated with the cytoplasmic face of the inner membrane. It has been previously suggested that MBP interacts with MalF and MalG during sugar transport across the inner membrane. In two independent genetic studies, reported here, residue 210 of MBP has been identified as an important site for its interaction with MalF. In one study, allele-specific suppressors of a malF mutation, malF506, were isolated and yielded mutations which altered residue tyrosine 210 of MBP to aspartic acid. In the other study, dominant mutations in malE (the structural gene of MBP) were isolated; one of these altered the same tyrosine residue (210) to cysteine. It was shown that the Y210C MBP mutant is also an allele-specific suppressor malF506, and that of the suppressor MBP alleles also exhibited dominant-negative phenotypes. Previously it was shown that alterations at residues glycine 13 and aspartate 14 of MBP can result in suppression of a malG mutant. From these results and those described, it is possible to propose a simple model in which the amino-terminal lobe of MBP interacts with MalG and the carboxy-terminal lobe of MBP interacts with MalF. The locations of residues 13, 14 and 210 on the three dimensional structure of MBP are in keeping with this model. PMID- 8411173 TI - Structural polymorphism of d(GA.TC)n DNA sequences. Intramolecular and intermolecular associations of the individual strands. AB - Alternating d(GA.TC)n sequences are highly structurally polymorphic. Most of their conformational flexibility is likely to reside in the structural properties of the individual strands themselves. In this paper the conformational behaviour of the d(GA)20 and d(TC)20 oligonucleotides was analysed. Formation of d(GA)20 intramolecular duplexes is observed at any pH value, from 8.3 to 4.6. On the other hand, intramolecular d(TC)20 duplexes are formed only under acidic conditions. The acid d(TC)20 intramolecular duplex is likely to be stabilized through the formation of C+C pairs, the thymine residues remaining unpaired. The d(GA)20 oligonucleotide also forms intermolecular duplexes which coexist with the intramolecular forms at any pH, from 8.3 to 4.6. The structural conformation adopted by the d(TC)20 oligonucleotide at neutral pH is uncertain. Under these conditions, this oligonucleotide shows an electrophoretic apparent molecular weight consistent with the formation of a bimolecular complex. However, no hydrogen bonding was observed to occur under these conditions. Implications of these results for an understanding of the molecular principles behind the conformational flexibility of alternating d(GA.TC)n sequences are discussed. The possible biological significance of these results is also discussed. PMID- 8411174 TI - Structural transitions during maturation of bacteriophage lambda capsids. AB - The three-dimensional structures of the procapsid and of the mature capsid of bacteriophage lambda were determined to a resolution of approximately 3.4 nm by cryo-electron microscopy and image processing. The mature lambda capsid contains two major proteins, gpE and gpD, arranged on a T = 7 lattice, with gpE arranged as hexamers and pentamers and gpD arranged as trimers. The hexamers and pentamers in the virion display a cartwheel-like structure, with skewed spokes (or arms) radiating out from a central hexameric hub. The thimble-shaped gpD trimers are superimposed on the trivalent interaction point of these arms. A reconstruction of a lambda D- mutant capsid to lower resolution shows no trace of these trimers, thus revealing the interactions of the underlying arms. The procapsid has elongated, irregularly shaped hexamers with gpE subunits set perpendicularly to the capsid surface. PMID- 8411175 TI - Residues of the Bacillus subtilis phage phi 29 transcriptional activator required both to interact with RNA polymerase and to activate transcription. AB - Regulatory protein p4 from Bacillus subtilis phage phi 29 activates transcription from the viral late promoter, PA3, by stabilizing the binding of RNA polymerase to the DNA as a closed complex. Protein p4-induced DNA bending and direct contacts between p4 and RNA polymerase have been proposed to play a role in P(A3) activation. By site-directed mutagenesis at the carboxyl end of protein p4 we have identified residues that are critical both to interact with RNA polymerase and to activate transcription. Substitution of arginine 120 gives rise to a p4 derivative unable to activate transcription, that can bind to DNA and induce a normal DNA bending, but does not stimulate the binding of RNA polymerase to the promoter and cannot form complexes with RNA polymerase. Modification of the closely located residue leucine 117 had a similar but milder effect. The results obtained suggest that arginine 120 and leucine 117 form part of the activating domain of the protein, and show that direct contacts between protein p4 and RNA polymerase play a critical role in transcription activation. The p4-induced DNA bending is therefore necessary but not sufficient for the activation of the PA3 promoter. PMID- 8411176 TI - The 2.5 A structure of pokeweed antiviral protein. AB - The pokeweed antiviral protein (PAP), isolated from the leaves of Phytolacca americana, is one of a family of plant and bacterial ribosome-inhibiting proteins (RIPs) which act as specific N-glycosidases on rRNA. Here we report the three dimensional structure of PAP determined to 2.5 A resolution by X-ray crystallography. After 14 rounds of refinement, the R factor is 0.17 for 5.0 to 2.5 A data. The protein is homologous with the A chain of ricin and exhibits a very similar folding pattern. The positions of key active site residues are also similar. We also report the 2.8 A structure of PAP complexed with a substrate analog, formycin 5'-monophosphate. As seen previously in ricin, the formycin ring is stacked between invariant tyrosines 72 and 123. Arg179 bonds to N-3 which is thought to be important in catalysis. PMID- 8411177 TI - A structural basis for sequence comparisons. An evaluation of scoring methodologies. AB - A residue-exchange matrix has been derived that is suitable for comparison of amino acid sequences. This matrix is based on the tabulation of 207,795 amino acid replacements observed in 65 homologous sets of structurally aligned three dimensional structures (235 proteins). The majority of the data is from structural comparisons where there is between 15 and 40% sequence identity. As a result, a scoring matrix such as the one devised here should provide a sensitive basis for the comparison of amino acid sequences and the search for homologous sequences in amino acid databases. In order to assess the value of this matrix we have made a comparative analysis with 12 other published scoring matrices that have been used for the alignment of protein amino acid sequences. We find that the matrix derived here is among the better performers in terms of alignment significance, detection of homologous sequences and the accuracy of alignments. PMID- 8411178 TI - How consistent are molecular dynamics simulations? Comparing structure and dynamics in reduced and oxidized Escherichia coli thioredoxin. AB - In this study we have examined several parameters that can be used for checking the consistency and accuracy of protein structures and molecular dynamics simulations. This is done by comparing: (1) three X-ray structures of oxidized Escherichia coli thioredoxin (Trx-S2); (2) 14 NMR structures of reduced E. coli thioredoxin (Trx-(SH)2); and (3) 30 different simulations, 15 of Trx-S2 and 15 of Trx-(SH)2. The energy, the agreement with NOE data, the root-mean-square deviation between structures, and the surface characteristics of all these structures are analyzed. The 30 simulations, four water simulations, 20 standard vacuum simulations and six alternative vacuum simulations, are examined with respect to mobility, temperature factors and aromatic side-chain mobility. It is shown that although vacuum simulations may reproduce some parameters, all the features of a water simulation cannot be reproduced in any of these simulations. Several of the parameters described above are shown to be good for discriminating between an accurate and an inaccurate simulation. It is also shown that 100 ps is too short a time to obtain statistically certain temperature factors and correlation functions of aromatic side-chain motions. The results also suggest that performing ten 100 ps simulations spans the conformation space better than one 1 ns simulation. PMID- 8411179 TI - Preliminary crystallographic study of D(-)-mandelate dehydrogenase from Rhodotorula graminis. AB - NAD+ dependent D(-)-mandelate dehydrogenase from the yeast Rhodotorula graminis strain KGX 39 has been crystallized in three different forms using the hanging drop vapour diffusion method at 15 to 20 degrees C. Type I crystals belong to space group P222(1), P22(1)2(1) or P2(1)2(1)2(1) with a = 100.3 A, b = 117.4 A, c = 80.4 A and are likely to contain a dimer in the crystallographic asymmetric unit. They diffract to dmin = 3.0 A. Type II crystals belong to space group P22(1)2(1) or P2(1)2(1)2(1) with a = 187.8 A, b = 122.9 A, c = 72.1 A and contain probably two dimers in the crystallographic asymmetric unit. They diffract to dmin = 1.8 A. Type III crystals belong to space group P2(1)2(1)2(1) with a = 109.6, b = 52.0 A, c = 145.7 A, and are likely to contain a dimer in the crystallographic asymmetric unit. They diffract at least to dmin = 2.5 A. PMID- 8411180 TI - Crystallization and preliminary crystallographic investigations of the soluble glucose dehydrogenase from Acinetobacter calcoaceticus. AB - Single crystals of the soluble glucose dehydrogenase (GDH) from Acinetobacter calcoaceticus have been grown by the vapour diffusion method. These crystals diffract to beyond 2.1 A and are suitable for X-ray crystallography. The space group was determined to be P2(1) with unit cell parameters a = 55.5 A, b = 104.5 A, c = 86.5 A and beta = 99.8 degrees. One asymmetric unit contains a dimer of the GDH molecule. PMID- 8411181 TI - Crystallization and preliminary X-ray diffraction studies of Bacillus stearothermophilus farnesyl diphosphate synthase expressed in Escherichia coli. AB - Thermostable farnesyl diphosphate synthase (EC 2.5.1.10) from Bacillus stearothermophilus, which was overexpressed in Escherichia coli, has been crystallized by the vapor-diffusion procedure. Tetragonal crystals were obtained using ammonium sulfate as a precipitant. The crystals diffracted X-rays to about 3 A resolution. The diffraction pattern indicated that the space group is I4(1)22 with unit-cell dimensions of a = b = 114 A and c = 247 A. It is thought that the asymmetric unit comprises two or three molecules of farnesyl diphosphate synthase. PMID- 8411182 TI - Structural characterization of the early indoleacetic acid-inducible genes, PS IAA4/5 and PS-IAA6, of pea (Pisum sativum L.). AB - Two early auxin-inducible genes (PS-IAA4/5 and PSIAA6) from pea were cloned using previously isolated complementary DNA sequences. They are present in single copy per haploid genome, and are members of a large divergent multigene family that encodes similar proteins. The genes were structurally characterized and sequence analysis of their 5'-flanking regions revealed the presence of several highly conserved sequences found in various auxin-regulated genes from other plant species. Their coding regions are interrupted by three and two introns, respectively. Introns two and three of PS-IAA4/5 and introns one and two of PS IAA6 are located in identical positions. These genes encode proteins of 189 (21,036 Da) and 179 (20,330 Da) residues that are 46% identical. They also share a significant degree of identity (42 to 80%) with other proteins encoded by auxin regulated genes in soybean, mungbean and Arabidopsis thaliana. All proteins contain four conserved domains ranging in size from 9 to 43 amino acids. Their most prominent feature is the presence of a highly charged N terminus consisting of two clusters of acidic residues separated by a cluster of basic amino acids. PMID- 8411183 TI - Molecular characterization of the DNA puff C-8 gene of Rhynchosciara americana. AB - We have mapped the only transcription unit known to be present in the C-8 DNA puff of Rhynchosciara americana and describe the isolation and sequence of a cDNA clone, pRa C-8-22, which contains a nearly complete copy of the mRNA transcribed from this DNA puff and part of the sequence of genomic clone BSC8-0.9, which contains the promotor region and the remainder of the transcription unit. The characteristics of the protein predicted from the ORF present in the cDNA indicate that it is unique and secreted. PMID- 8411184 TI - Electromyographic and kinematic studies of tail movements in dogs during treadmill locomotion. AB - Electromyographic (EMG) activities of three tail muscles, the extensor caudae lateralis (ECL), abductor caudae externus (ACE), and flexor caudae longus (FCL), were recorded bilaterally in seven adult dogs during walking, trotting, and galloping on a treadmill. Each dog's movements were recorded with a 16 mm high speed camera system, and angular movements of the tail were analyzed. During walking and trotting, reciprocal EMG bursts were observed between right and left tail muscles and corresponded with lateral movements of the tail. The tonic discharges that were observed in ECL and FCL seemed to maintain the position of the tail. During galloping, synchronized EMG activity of all tail muscles produced reactive torques to counter those generated by cyclic limb movements and kept the tail in a stable position. These results suggest that tail movements are important in maintaining body balance during locomotion in the dog. PMID- 8411185 TI - Spermathecal cytology of Ambystoma opacum (Amphibia: Ambystomatidae) and the phylogeny of sperm storage organs in female salamanders. AB - Sperm storage glands, spermathecae, were examined from mated female Ambystoma opacum during the breeding season. No differences occur in the spermathecal ultrastructure of individuals sacrificed prior to oviposition and those sacrificed within 3 days of removal from tended clutches of recently oviposited eggs. The simple tubuloalveolar glands produce two types of secretory vacuoles. Apical secretory vacuoles contain glycosaminoglycans for export into the lumen to bathe stored sperm, perhaps providing the chemical/osmotic environment necessary for sperm quiescence. The other type of secretory vacuole contains an unsaturated lipid that is produced for export into the connective tissue surrounding the spermathecae. The role of this secretion may involve the contraction of myoepithelial cells, resulting in sperm expulsion. Some sperm undergo degradation in the spermathecal epithelium, and an interepithelial leukocyte was observed in one specimen. Apical secretory vacuoles and sperm are absent from the spermathecae of a specimen sacrificed 62 days after removal from a tended egg clutch. This is the first report on the spermathecal cytology of a salamander from the Ambystomatidase, and comparisons with salamanders from other families provide a morphological basis for considering spermathecae polyphyletic within the Caudata. PMID- 8411186 TI - Scaling of cursoriality in mammals. AB - Data on limb bone lengths from 64 mammalian species were combined with data on 114 bovid species (Scott, '79) to assess the scaling of limb lengths and proportions in mammals ranging from 0.002 to 364 kg. We analyzed log-transformed data using both reduced major axis and least-squares regression to focus on the distribution across mammals of two key traits-limb length and metatarsal/femur ratio--associated with cursorial adaptation. The total lengths of both fore and hindlimbs scale in a manner very close to the M0.33 predicted by geometric similarity. Thus the relative limb lengths of large mammals, including bovids, generally regarded among the most cursorial of mammals, are very similar to those of the rodents and insectivores in this sample. Metatarsal/femur ratio also shows little change with changing mass, although bovids tend to have relatively longer metapodials than do other families in the sample. We argue that many of the remaining morphological traits associated with cursoriality (e.g., reduction in joint mobility and number of distal limb bone elements) promote cursoriality only at large body sizes. These results lead us to question the general perception that cursoriality is most widespread among large mammals. We also suggest that discussions of cursoriality should focus explicitly on the two partially independent aspects of performance that are otherwise confounded under this general term--speed and the ability to cover substantial distance. PMID- 8411187 TI - Histomorphometry of immature rat testis after heating. AB - The scrotal testes of albino rats aged 35 and 45 days were immersed in water at constant temperatures of 43 degrees C, 44 degrees C, or 45 degrees C for periods of 15-45 min in a special heating device. At an age of 60 days, the rats were mated in individual cages with two primiparous rats each. At an age of 90 days, they were killed and their testes were histologically processed. Rats with testes that had been subjected to heating when the animals were 45 days old showed both alterations of the seminiferous tubules and a decrease in fertilizing capacity. The effect of heat was greater in animals at 45 than at 35 days of age. In heat treated testes, tubules contained PAS-positive concretion, sometimes engulfed by macrophage-derived giant cells and multinucleate cells derived from spermatids that failed to separate during spermiogenesis. The decrease in testicular volume observed after heat treatment was due mainly to reduced parenchymal volume. Thermic lability of seminiferous stem cells increases with age until adulthood, and recovery from heat injury declines. PMID- 8411188 TI - How does the Na(+)-Ca2+ exchanger work in the intact cardiac cell? PMID- 8411189 TI - Is "fuzzy space" necessary for Ca2+ extrusion on the Na(+)-Ca+ exchanger in cardiac myocytes? PMID- 8411190 TI - Synthesis of leukotrienes by freshly harvested endothelial cells. AB - Endothelial cells produce endothelin, a powerful vasoconstrictor. We report the release of additional vasoconstrictor material in conditional filtrate from freshly harvested cells, which we identified as leukotrienes by radioimmunoassay (RIA) and by high pressure liquid chromatography (HPLC). The material was collected in cell free filtrates by superfusion of freshly harvested bovine endothelial cells attached to cytodex-3 microcarrier beads. Cells and beads form a dense network on filter paper permitting collection of cell free filtrate. The amount of leukotrienes in conditioned filtrate was 158 +/- 21 picograms/million cells. The calcium ionophore A23187 stimulated the release of leukotrienes (392.0 +/- 47.6). The peak of leukotriene production occurred within an hour after incubation of cells slowly declining thereafter. Conditioned filtrate to which indomethacin had been added caused coronary vasoconstriction in the perfused rat heart preparation, as did synthetic leukotrienes C4, D4 and E4. It was found by RIA and HPLC that some of the constrictor effect of conditioned filtrate derived from leukotrienes. PMID- 8411191 TI - Pertussis toxin unmasks stimulatory myocardial A2-adenosine receptors on ventricular cardiomyocytes. AB - Pertussis toxin-pretreatment abolished the contractility- and cAMP-decreasing effects of the A1-adenosine receptor agonist (-)-N6-phenylisopropyladenosine (R PIA) in the presence of isoprenaline in isolated ventricular cardiomyocytes from guinea-pigs, indicating that these stimulatory effects of A1-adenosine receptors are mediated via pertussis toxin-sensitive G-proteins. Furthermore, the decrease in contractile response by the A1/A2-adenosine receptor agonist 5'-N ethylcarboxamidadenosine (NECA) was abolished. Moreover, NECA increased cAMP content in pertussis toxin-pretreated cells. Thus, pertussis toxin unmasked cAMP augmenting effects of NECA, indicating that NECA can stimulate A2-adenosine receptors on cardiomyocytes. Thereby, the present study provides evidence that besides cAMP- and contractility-decreasing A1-adenosine receptors, cAMP increasing A2-adenosine receptors coexist on ventricular cardiomyocytes, which do not influence contractile response. PMID- 8411192 TI - Stimulation of opioid receptors on cardiac ventricular myocytes reduces L type Ca2+ channel current. AB - Recent studies have indicated that opioid peptide receptors are present on cardiac ventricular cells and that Leucine enkephalin (LE), a naturally occurring delta opioid peptide receptor agonist, leads to marked reductions in twitch amplitude and in the cytosolic Ca2+ transient (Ca(i)) of single adult rat ventricular myocytes. The specific mechanisms by which Ca(i) is reduced by LE have not been fully elucidated. Specifically, it is unknown whether LE affects the Ca2+ current (ICa) of L type Ca2+ channels. In the present study we determined the effect of LE on ICa of individual cardiac ventricular cells freshly isolated from adult rats. LE (10(-8) M) decreased the amplitude of ICa by 40% (during regular whole cell voltage clamp depolarizations to 0 mV at 0.5 Hz at 23 degrees C from a holding potential of -40 mV). The relative magnitude of this effect increased with the magnitude of the test potential from -20 to +50 mV. ICa inactivation was also prolonged by LE. These effects of LE on ICa were abolished by Naloxone (NAL), an opioid receptor antagonist. Thus, the effects of the opioid peptide, LE, to decrease the Ca(i) transient and contraction amplitudes in individual cardiac ventricular cells, are, in part, mediated by an LE induced reduction in ICa. PMID- 8411193 TI - Metabolism in non-ischemic myocardium during coronary artery occlusion and reperfusion. AB - Blood flow and metabolism in non-ischemic myocardium were studied at baseline and during occlusion and reperfusion of the left anterior descending coronary artery in closed chest dogs using positron emission tomography. Myocardial blood flow (MBF) and oxygen consumption (MVO2) in non-ischemic tissue were each increased by 28% relative to the rate pressure product during occlusion, consistent with increased work to compensate for the dyskinetic segment. MVO2 in non-ischemic sectors remained elevated relative to the rate pressure product early (1-2 h) post-reperfusion, 21% above baseline, but subsequently normalized. When sectors with normal blood flow during occlusion were divided into sectors adjacent to and remote from the risk zone, MBF in the 2 sector groups was similar at all times, but metabolic differences were found. MVO2 was depressed by 15% in adjacent relative to remote sectors 1 day post-reperfusion, with a concomitant 62% increase in glucose metabolic rate; relative increases in glucose metabolism were found only when glucose metabolism was low in remote myocardium, suggesting a decreased suppressibility of glucose metabolism in adjacent myocardium. The kinetics of (1-11C] palmitate were also altered in adjacent sectors, consistent with a small increase in esterification relative to oxidation of long chain fatty acids. Thus, sectors adjacent to ischemic segments show metabolic changes similar to those seen in reversibly injured post-ischemic tissue, despite normal blood flow during occlusion. PMID- 8411194 TI - Pacing-induced ventricular fibrillation leading to oxygen free radical formation in aerobically perfused rat hearts. AB - The objective of this study was to determine whether electrically-induced ventricular fibrillation can elicit oxygen free radical formation, even in the absence of ischemia and reperfusion. Rat hearts (n = 8 in each group) were perfused aerobically at 37 degrees C and ventricular fibrillation was induced by pacing (20 Hz, 1200 beats/min) for 10 min, during which time no changes in coronary flow rates were observed. In this study, electron spin resonance (ESR) studies using the spin trap 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) demonstrated the formation of oxygen free radicals consisting of 1:2:2:1 quartet with peak concentration at the 3rd min of fibrillation in non-ischemic electrically fibrillating hearts. Since there were no significant changes in coronary flow rates during ventricular fibrillating (22.3 +/- 1.3 ml/min in control vs 22.3 +/- 0.9 ml/min in pacing-induced fibrillating group), the formation of oxygen free radicals could not be attributed to a pacing-induced ischemic event. In additional studies, hearts were paced by 10 Hz (600 beats/min), to demonstrate whether ventricular tachycardia could elicit free radical formation. In these experiments the genesis of oxygen free radicals was not observable after 1 min, 5 min, and 10 min of tachycardia, but a small amount of OH. radicals was detected at the 3rd min of ventricular tachycardia. When DMPO was infused into the heart giving a final perfusate concentration of 2.5 mmoles/litre during the pacing induced fibrillation period, myocardial function (aortic flow, cardiac output, left ventricular developed pressure, first derivative of left ventricular developed pressure, and end-diastolic pressure) was significantly improved after the postfibrillating period. In conclusion, our studies clearly show that electrically-induced ventricular fibrillation is capable of eliciting free radical formation even in the absence of ischemia and reperfusion, and the cardioprotective effect of the spin trap is directly originated from its free radical trapping property and not from the other pharmacological activities of DMPO. PMID- 8411195 TI - Implications of the coronary vascular endothelium as mediator of the vasodilatory and dromotropic actions of adenosine. AB - Recent studies have shown that the vasodilatory response of adenosine is partially endothelium dependent. Therefore, to further characterize the physiologic role of the vascular endothelium as mediator of adenosine's cardiovascular effects, we have examined in isolated perfused guinea-pig hearts the dromotropic and vascular effects of adenosine in the presence of the adenosine antagonist, XAC, covalently conjugated to latex microspheres of 0.07 microns diameter. Our results demonstrate that intravascular infusion of the microsphere XAC conjugates abolishes the vasodilatory and negative dromotropic effects of infused adenosine, and inhibits the dromotropic effects of hypoxia. As these particles because of their size remain intravascularly confined, we conclude that the dromotropic and vasodilatory effects of exogenous adenosine, and the dromotropic effects of hypoxia (endogenous adenosine), arise from the intravascular adenosine compartment acting by way of the vascular endothelium. In addition, while neither the temporal course of vasodilation nor the steady state of vasodilation caused by hypoxia were influenced by unconjugated XAC, our results do show that the microsphere XAC conjugate increases the time necessary for maximum vasodilation to occur during hypoxia. PMID- 8411196 TI - A comparative study of effects of isoproterenol and dihydroouabain on calcium transients and contraction in cultured rat ventricular cells. AB - We investigated the effects of isoproterenol, a beta-adrenergic agonist, and dihydroouabain, a Na+,K(+)-pump inhibitor, on Ca2+ transients and contraction of cultured rat ventricular cells and compared the effects with those of altered external ion concentrations, with special reference to the changes in diastolic intracellular free calcium concentration ([Ca2+]i). We measured [Ca2+]i of cultured cell aggregates, stimulated at 1.0 Hz, with the use of dual-wavelength microfluorometry of fura-2, at room temperature (24-26 degrees C). The contraction was measured as a shortening of the aggregates using a photodiode array placed on a video monitor. Isoproterenol increased the magnitude of contraction and the peak amplitude of the Ca2+ transient, in a concentration (10( 9)-10(-6) M)-dependent manner, but did not change the diastolic Ca2+ level. Isoproterenol at 10(-7) M or higher significantly shortened the duration of contraction and half decay time of a Ca2+ transient yet it did not change the time to peak. Dihydroouabain (10(-7)-10(-5) M) increased the contraction and elevated both systolic and diastolic calcium levels but it did not alter the duration of contraction, the time to peak and the half decay time. The effects of dihydroouabain on Ca2+ transients were mimicked by lowering [K+]o (0.4 mM), by lowering [Na+]o (74 mM) or by elevating [Ca2+]o (3.6 or 5.4 mM). Ryanodine (10( 5) M), by itself, decreased systolic Ca2+ transient amplitude, increased diastolic Ca2+ levels and prolonged the time to peak and the half decay time. In the presence of ryanodine, isoproterenol increased both systolic and diastolic [Ca2+]i. Thus, most procedures that increased the systolic Ca2+ transient amplitude increased the diastolic Ca2+ levels as well, and enhanced the contraction. The only exception was isoproterenol that markedly increased the systolic Ca2+ transient amplitude without affecting the diastolic Ca2+ level, a finding in keeping with the observation that isoproterenol stimulates Ca2+ uptake by the sarcoplasmic reticulum. PMID- 8411197 TI - Characterization of a Mg-dependent, Na-inositol co-transport process in cardiac sarcolemmal vesicles. AB - [3H]-myo-Inositol transport into cardiac sarcolemmal vesicles is both a time- and Na(+)-dependent process. Transport was stimulated 3-fold by 1 microM valinomycin suggesting the process is electrogenic. myo-Inositol transport was dependent upon the presence of Mn2+ or Mg2+ but not Ca2+. Kinetic analysis revealed a high affinity transport process that was saturable (Vmax, 650 +/- 71 pmoles myo inositol/mg protein/min; Km, 37.7 +/- 1.3 microM) and a low affinity process that was unsaturable up to 1.0 mM substrate. Transport was not inhibited by 1.0 mM of either L-glucose, D- or L-galactose. However, D-glucose and L-fucose at 1.0 mM were inhibitory. Higher concentrations (30 mM) of each of the sugars inhibited transport. myo-Inositol transport was also inhibited by the inositol isomers (1.0 mM), D-chiro-inositol, epi-inositol and scyllo-inositol with scyllo-inositol being most effective. Kinetic analysis established scyllo-inositol as a competitive inhibitor of cardiac myo-inositol transport, increasing the Km for substrate three-fold (122 +/- 21 microM). These data indicate that inositol transport across cardiac sarcolemma is a Mg(2+)-dependent Na+ co-transport process that is electrogenic and stereospecific. PMID- 8411198 TI - Early postnatal development of contractile performance and responsiveness to Ca2+, verapamil and ryanodine in the isolated rat heart. AB - Contractile performance of the isolated perfused rat heart and its inotropic response to Ca2+, verapamil and ryanodine was studied in 1-, 2-, 4-, 7- and 12 day-old animals. Values of the developed force revealed two different phases: a slow decrease from day 1 to day 4 followed by a steep increase up to day 12. A similar biphasic time course was observed in the magnitude of the inotropic effect of Ca2+ (0.6-10.0 mmol.l-1): decrease from day 1 to day 4 followed by an increase up to day 7. The sensitivity to Ca2+ was, however, not changed. An analogous biphasic response was also observed during perfusion with the calcium antagonist verapamil (10(-9) to 3.3 x 10(-7) mol.l-1): the sensitivity to negative inotropic effect rose from day 1 to day 4 and then decreased at day 7 (values of IC50 were 170 +/- 61, 17 +/- 6 and 171 +/- 60 10(-9) mol.l-1 S.E.M. on day 1, 4 and 7, respectively). The contractile response to the inhibitor of calcium release from sarcoplasmic reticulum (SR)--ryanodine (10(-6) mol.l-1)--was surprisingly high already in 1-day-old animals (inhibition of contraction by more than 50%) indicating the presence of functionally active SR in the rat heart just after birth. Our data clearly shows that the early development of contractile function and inotropic responsiveness of the rat heart is not linear and changes dramatically during the first week of life. PMID- 8411199 TI - Compositional properties of coding sequences and mammalian phylogeny. AB - The compositional distributions of large DNA fragments reflect those of the isochores that make up vertebrate genomes and can provide novel phylogenetic insights in the case of mammalian genomes (see Sabeur et al. 1993). This approach has been complemented here by an analysis of the compositional patterns of coding sequences and their codon positions (which also reflect the isochore pattern) and by a comparison of the base compositions of codon positions from homologous genes in a number of pairs of species. The results obtained using these two approaches support the existence of a general compositional pattern for mammalian genomes and of a distinct pattern for Myomorpha. The other two "special" patterns identified in a megachiropteran and in pangolin could not be tested here. PMID- 8411200 TI - The distribution of genes on chromosomes: a cytological approach. AB - Studies during the last 20 years have shown that the chromosomes of many organisms, especially those of higher vertebrates, consist of a series of segments having different properties. These can be recognized as, for example, G- and R-bands. Recent studies have indicated that genes tend to lie in the R-bands rather than in the G-bands, although the number of genes that has been mapped with high precision is, as yet, only a very small proportion of the total, probably much less than 1%. We have therefore sought to study the distribution of genes on chromosomes using a cytological approach in conjunction with "universal" markers for genes. Such markers include mRNA and the gene-rich, G+C-rich H3 fraction of DNA, both of which can be localized using in situ hybridization, and DNase I hypersensitivity, and digestion by restriction enzymes known to show selectivity for the CpG islands associated with active genes, both of which can be detected using in situ nick translation. We have chosen to use the approaches involving in situ nick translation and have shown that the patterns of DNase I hypersensitivity and of CpG islands on human chromosomes show a strict correspondence to R-banding patterns: Deviations from R-banding patterns reported by previous investigators who have made similar studies appear to be attributable to excessive digestion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411201 TI - The frequency of two-base tracts in eukaryotic genomes. AB - The frequency of two-base tracts is surveyed in a wide range of eukaryotic genomes using the special program TRACTS. All three two-base families are surveyed: R.Y (A,G.C,T), K.M (A,C.G,T), and S;W (A.T and G.C). Data for the human beta-globin complex, for the tobacco chloroplast, and for 247 nt mammalian promoter regions are presented. All two-base tracts longer than three or four bases are overrepresented to an extent surpassing by far their occurrence in a randomized DNA population in the majority of the genomic regions analyzed; 20-30 long tracts are quite frequent, against the statistical odds. R.Y tracts are found at the largest excess, K.M tract to a slightly lesser extent, while S.W tracts are found at a moderate yet significant excess. The majority of the tracts manifest only a limited extent of tandem repeat structures. The idea that the two base tracts serve as unwinding elements is considered. PMID- 8411202 TI - Dinucleotides and G+C content in human genes: opposite behavior of GpG, GpC, and TpC at II-III codon positions and in introns. AB - We have studied the behavior of the dinucleotide preferences under G+C content variation in human genes. The doublet preferences for each dinucleotide were compared between two functionally distinct zones in genes, the II-III codon positions, and the introns. The 16 dinucleotides have been tentatively classified in three groups: AA, AC, CC, CT, and GA, doublets showing no difference between introns and II-III codon positions in the full range of G+C variation TG and TA, which differ in the full range of G+C variation AT, AG, GT, TC, TT, GG, GC, CG, and CA, which show differences in regions over 50% G+C A remarkable pattern observed concerns the behavior of GG, GC, and TC, which showed opposite trends in II-III codon positions and in introns. If codon positions and introns are under the same structural requirements and the same mutational bias, our results indicate that the differences observed could be related to post-transcriptional constraints acting on mRNA. PMID- 8411203 TI - Directional mutation pressure, mutator mutations, and dynamics of molecular evolution. AB - Using a general form of the directional mutation theory, this paper analyzes the effect of mutations in mutator genes on the G+C content of DNA, the frequency of substitution mutations, and evolutionary changes (cumulative mutations) under various degrees of selective constraints. Directional mutation theory predicts that when the mutational bias between A/T and G/C nucleotide pairs is equilibrated with the base composition of a neutral set of DNA nucleotides, the mutation frequency per gene will be much lower than the frequency immediately after the mutator mutation takes place. This prediction explains the wide variation of the DNA G+C content among unicellular organisms and possibly also the wide intragenomic heterogeneity of third codon positions for the genes of multicellular eukaryotes. The present analyses lead to several predictions that are not consistent with a number of the frequently held assumptions in the field of molecular evolution, including belief in a constant rate of evolution, symmetric branching of phylogenetic trees, the generality of higher mutation frequency for neutral sets of nucleotides, the notion that mutator mutations are generally deleterious because of their high mutation rates, and teleological explanations of DNA base composition. PMID- 8411204 TI - Potential problems in estimating the male-to-female mutation rate ratio from DNA sequence data. AB - It is commonly believed that the rate of mutation is much higher in males than in females because the number of germ-cell divisions per generation is much larger in males than in females. However, the precise magnitude of the male-to-female mutation rate ratio (alpha m) remains unknown. Recently there have been efforts to estimate alpha m by using DNA sequence data from different species. We have studied the potential problems in such an approach. We found that the rate of synonymous substitution varies about fivefold among X-linked genes, as large as the variation among autosomal genes. This large variation makes the assumption of selective neutrality of synonymous changes dubious, so one should be cautious in using the synonymous rates in X-linked and autosomal genes to estimate alpha m. A similar difficulty was also observed in using nonhomologous intron sequences to estimate alpha m. Contrary to the expectation that X-linked sequences should evolve more slowly than autosomal sequences, the Alu repeat in the last intron of the X-linked zinc finger gene has evolved faster than the four autosomal Alu repeats used in this study. It appears that the best way to estimate alpha m is to use homologous sequences. However, such sequences may be involved in gene conversion events. In fact, we found evidence that the Y-linked and X-linked zinc finger genes have been involved in multiple conversion events during primate evolution. Thus, the possibility of gene conversion should be considered when using homologous sequences to estimate alpha m. PMID- 8411205 TI - A model for the mechanism of initial generation of short interspersed elements (SINEs). AB - Most animal genomes contain a large number of short interspersed elements (SINEs) that have a composite structure and contain a region that is homologous to a tRNA. The majority of SINEs have been found to be derived from a tRNA(Lys), being categorized as members of a superfamily of tRNA(Lys)-related SINEs. The consensus sequences of five SINEs that belong to this superfamily were aligned. It was found that, in the tRNA-unrelated region, there are two sequence motifs that are almost identical among these five SINEs and are at a distance of 10-11 nucleotides from each other. This observation suggests a common evolutionary origin of these SINEs and/or some function(s) for these motifs. Similar sequences were unexpectedly found to be present in the sequences complementary to the U5 regions of several mammalian retroviruses whose primer is a tRNA(Lys). On the basis of these findings, we propose a possible model for the generation of SINEs whereby they are derived from a "strong stop DNA" with a primer tRNA that is an intermediate in the process of reverse transcription of certain retroviruses. PMID- 8411206 TI - Organization of the variant domains of alpha satellite DNA on human chromosome 21. AB - The de novo creation of long, homogeneous, satellite DNA domains was postulated previously to occur by saltatory amplification. In this paper, pulsed field gel electrophoresis analysis of the alpha satellite DNA block organization of the human chromosome 21 supports this hypothesis. Double-dimension electrophoresis indicated that the variant copies of the basic alpha satellite repeat of chromosome 21 are organized in a single 3,150 Kb-long domain. It was also established that the other satellite DNAs found in man (beta, II, and III) are organized independently of the alpha satellite DNA block of the same chromosome. PMID- 8411207 TI - Multiple L1 progenitors in prosimian primates: phylogenetic evidence from ORF1 sequences. AB - One of the uncertainties regarding the evolution of L1 elements is whether there are numerous progenitor genes. We present phylogenetic evidence from ORF1 sequences of slow loris (Nycticebus coucang) and galago (Galago crassicaudatus) that there were at least two distinct progenitors, active at the same time, in the ancestor of this family of prosimian primates. A maximum parsimony analysis that included representative L1s from human, rabbit, and rodents, along with the prosimian sequences, revealed that one of the galago L1s (Gc11) grouped very strongly with the slow loris sequences. The remaining galago elements formed their own unique and strongly supported clade. An analysis of replacement and silent site changes for each link of the most parsimonious tree indicated that during the descent of the Gc11 sequence approximately two times more synonymous than nonsynonymous substitutions had occurred, implying that the Gc11 founder was functional for some time after the split of galago and slow loris. Strong purifying selection was also evident on the galago branch of the tree. These data indicate that there were two distinct and contemporaneous L1 progenitors in the lorisoid ancestor, evolving under purifying selection, that were retained as functional L1s in the galago lineage (and presumably also in the slow loris). The prosimian ORF1 sequences could be further subdivided into subfamilies. ORF1 sequences from both the galago and slow loris have a premature termination codon near the 3' end, not shared by the other mammalian sequences, that shortens the open reading frame by 288 bp. An analysis of synonymous and nonsynonymous substitutions for the 5' and 3' portions, that included intra- and inter subfamily comparisons, as well as comparisons among the other mammalian sequences, suggested that this premature stop codon is a prosimian acquisition that has rendered the 3' portion of ORF1 in these primates noncoding. PMID- 8411208 TI - Generation of VNTRs and heteroplasmy by sequence turnover in the mitochondrial control region of two elephant seal species. AB - We describe an unusual repetitive DNA region located in the 3' end of the light (L)-strand in the mitochondrial control region of two elephant seal species. The array of tandem repeats shows both VNTR (variable-number tandem repeat) and sequence variation and is absent from 12 compared mammalian species, except for the occurrence in the same location of a distinct repetitive region in rabbit mtDNA and a similar repeat in the harbor seal. The sequence composition and arrangement of the repeats differ considerably between the northern elephant seal (Mirounga angustirostris) and the southern species (M. leonina) despite an estimated divergence time of 1 MY (based on an mtDNA-RNA gene and the nonrepetitive control region). Analysis of repeat sequence relationships within and between species indicate that divergence in sequence and structure of repeats has involved both slippage-like and unequal crossingover processes of turnover, generating very high levels of divergence and heteroplasmy. PMID- 8411209 TI - Sequence comparison and mutational analysis of elements that may be involved in the regulation of DNA synthesis in HIV-1. AB - The large number of sequenced clones of HIV-1 and related viruses made it possible to indicate conserved elements with potential regulatory or structural functions. Such analysis was combined with directed mutagenesis in order to investigate the importance of elements that may influence the initiation of plus strand DNA synthesis. The main site for plus-strand initiation is a polypurine tract near the 3' end of the viral RNA (the 3' PPT). An exact copy of this PPT is located in the middle of the genome (the internal PPT). Upstream from the internal PPT there is an inverted repeat. Mutants designed to upset the internal PPT (i.e., purine to pyrimidine changes), as well as mutants designed to abolish the potential stem-loop formation (changes around the internal PPT or in the upstream inverted repeat) both resulted in viruses with a reduced ability to replicate. Upsetting the stem-loop formation was, however, less harmful than changing the polypurine nature of the PPT. Changing a conserved T on the 3' side of the PPT to a C did not affect the phenotype. PMID- 8411210 TI - Phylogenetic relationships of HMG box DNA-binding domains. AB - HMG boxes were initially identified as DNA-binding domains of the human RNA polymerase I (pol I) transcription factor hUBF and the animal high-mobility-group (HMG) protein family HMG1. Since then, numerous sequences of HMG-box-containing HMG proteins and other DNA-binding proteins from several species have become available. By sequence comparisons of a selected range of HMG boxes from these proteins and the construction of phylogenetic trees we show that the HMG box is highly conserved between DNA-binding proteins of organisms from all three eukaryotic kingdoms and that HMG boxes are linked by distinct evolutionary relationships. In addition, most HMG boxes display comparable hydropathy profiles and amino acid arrangements, which could serve as nuclear targeting sequences. PMID- 8411211 TI - Silk moth chorion pseudogenes: hallmarks of genomic evolution by sequence duplication and gene conversion. AB - The part of the genetic locus of the domesticated silk moth, Bombyx mori, in which high cysteine (Hc) chorion genes of late developmental specificity reside contains regions encompassing gene-like sequences which exhibit properties distinct from those of functional Hc genes. One of these regions has been characterized and shown to contain a chorion pseudogene, psi HcB.15, which shares pronounced similarities with a transcribed chorion pseudogene, psi HcB.12/13, which was characterized previously. Both pseudogenes are homologous to HcB chorion genes but bear multiple single nucleotide substitutions and short segmental mutations (insertions and deletions) which introduce translational frame shifts and termination codons in the coding regions. Structural characteristics unique to the two pseudogenes suggest that psi HcB.15 was generated first from a functional HcB gene and gave rise subsequently to psi HcB 12/13 as a result of a sequence duplication event. The two pseudogenes can be distinguished from each other by the presence of distinct regions of similarity to the consensus sequence of functional HcB genes which appear to have arisen from gene-conversion-mediated correctional events. These findings lend support to the hypothesis that chorion pseudogene sequences represent reservoirs of genetic information that participates in the evolution of the chorion locus rather than relics of inactivated genes passively awaiting extinction. PMID- 8411212 TI - Reconstructing evolution from eukaryotic small-ribosomal-subunit RNA sequences: calibration of the molecular clock. AB - The detailed descriptions now available for the secondary structure of small ribosomal-subunit RNA, including areas of highly variable primary structure, facilitate the alignment of nucleotide sequences. However, for optimal exploitation of the information contained in the alignment, a method must be available that takes into account the local sequence variability in the computation of evolutionary distance. A quantitative definition for the variability of an alignment position is proposed in this study. It is a parameter in an equation which expresses the probability that the alignment position contains a different nucleotide in two sequences, as a function of the distance separating these sequences, i.e., the number of substitutions per nucleotide that occurred during their divergence. This parameter can be estimated from the distance matrix resulting from the conversion of pairwise sequence dissimilarities into pairwise distances. Alignment positions can then be subdivided into a number of sets of matching variability, and the average variability of each set can be derived. Next, the conversion of dissimilarity into distance can be recalculated for each set of alignment positions separately, using a modified version of the equation that corrects for multiple substitutions and changing for each set the parameter that reflects its average variability. The distances computed for each set are finally averaged, giving a more precise distance estimation. Trees constructed by the algorithm based on variability calibration have a topology markedly different from that of trees constructed from the same alignments in the absence of calibration. This is illustrated by means of trees constructed from small-ribosomal-subunit RNA sequences of Metazoa. A reconstruction of vertebrate evolution based on calibrated alignments matches the consensus view of paleontologists, contrary to trees based on uncalibrated alignments. In trees derived from sequences covering several metazoan phyla, artefacts in topology that are probably due to a high clock rate in certain lineages are avoided. PMID- 8411213 TI - The isochore patterns of mammalian genomes and their phylogenetic implications. AB - The compositional distributions of high molecular weight DNA fragments from 20 species belonging to 9 out of the 17 eutherian orders were investigated by analytical CsCl density gradient centrifugation and by preparative fractionation in Cs2SO4/BAMD density gradients followed by analysis of the fractions in CsCl. These compositional distributions reflect those of the isochores making up the corresponding genomes. A "general distribution" was found in species belonging to eight mammalian orders. A "myomorph distribution" was found in Myomorpha, but not in the other rodent infraorders Sciuromorpha and Histricomorpha, which share the general distribution. Two other distributions were found in a megachiropteran (but not in microchiropteran, which, again, shares the general distribution) and in pangolin (a species from the only genus of the order Pholidota), respectively. The main difference between the general distribution and all other distributions is that the former contains sizable amounts (6-10%) of GC-rich isochores (detected as DNA fragments equal to, or higher than, 1.710 g/cm3 in modal buoyant density), which are scarce, or absent, in the other distributions. This difference is remarkable because gene concentrations in mammalian genomes are paralleled by GC levels, the highest gene concentrations being present in the GC richest isochores. The compositional distributions of mammalian genomes reported here shed light on mammalian phylogeny. Indeed, all orders investigated, with the exception of Pholidota, seem to share a common ancestor. The compositional patterns of the megachiropteran and of Myomorpha may be derived from the general pattern or have independent origins. PMID- 8411214 TI - Imidazole-substituted analogues of TRH limit behavioral deficits after experimental brain trauma. AB - Treatment with thyrotropin releasing hormone (TRH) or TRH analogues improves outcome after experimental brain or spinal cord trauma. TRH analogues with modifications at the N-terminal position of the tripeptide are effective, whereas analogues with modifications of the C-terminal residue are not. Imidazole substituted TRH analogues, which modify the middle amino acid (histidine) of the tripeptide, have more recently been developed but have not been evaluated in models of central nervous system (CNS) trauma. In the present studies two imidazole-substituted analogues--4(5)-NO2(Im)TRH and 2,4 diiodo(Im)TRH--are shown to improve behavioral recovery following fluid percussion-induced traumatic brain injury (TBI) in rats. Because 4(5)-NO2(Im)TRH has little endocrine activity and 2,4 diiodo(Im)TRH has minimal cardiovascular effects, these experiments support the hypothesis that the neuroprotective actions of TRH analogues are independent of their endocrine or autonomic actions. PMID- 8411215 TI - A nonpsychotropic cannabinoid, HU-211, has cerebroprotective effects after closed head injury in the rat. AB - HU-211 is a synthetic, nonpsychotropic cannabinoid, which has been shown to act as a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist. The cerebroprotective effects of this compound were assessed in a model of closed head injury in rats. Head trauma (HT) was induced in ether-anesthetized rats by a weight-drop device; recovery was followed up to 48 h. The clinical status of the rats was evaluated at 1, 24, and 48 h after injury, and the extent of edema formation was determined by specific gravity (SG) and water content measurements at 24 or 48 h. The integrity of the blood-brain barrier (BBB) was investigated using Evans-Blue extravasation at 4 h after HT. HU-211 at a dose of 25 mg/kg in middle-chain triglycerides (MCT) oil was given intraperitoneally immediately and 1, 2, or 3 h after impact, and its effect on the various parameters was studied. The drug was found to be very effective in improving motor function recovery. When the drug was given 1 h after HT, the percent of rats able to perform beam walking task on 8.5- and 5-cm wide beams was increased from 30% and 0% to 79% (p = 0.0172) and 57% (p = 0.0029), respectively. The percent of rats able to balance on a 1.5-cm beam for 20 and 40 sec was also significantly increased, from 9% and 0% to 72% (p = 0.0037) and 50% (p = 0.078), respectively. The drug was also effective in reducing the BBB breakdown by more than four fold, as compared with control (548 +/- 94 versus 128 +/- 19 ng Evans blue/g tissue; p < 0.05) and attenuating cerebral edema. SG was 1.0367 +/- 0.0007 versus 1.0399 +/- 0.0005, and percent water content was 83.06 +/- 0.57 versus 80.78 +/- 0.36 (p < 0.05) in control and HU-211 treated rats, respectively. Similar significant protection was found when the drug was injected 2 h after the injury; however, at 3 h the effect was somewhat less pronounced. We suggest that this novel drug is a potential cerebroprotector in head trauma with a therapeutic window of at least 2 to 3 h. PMID- 8411216 TI - Early assessment of neurologic deficits in the fluid percussion model of brain injury. AB - This study was designed to quantify the early neurologic effects of brain injury elicited by fluid percussion to the dura of cats. Propofol was used for surgical anesthesia because recovery in normal animals from an intravenous infusion was found to be nearly complete within 2 h of cessation and absolutely complete by 4 h. In addition, a cat coma scale (CCS) was developed that reflects normal (CCS, 14) to moribund (CCS, 3) behavior. The CCS values at 6 h were compared with the force of injury expressed in atmospheres (atm), maximum blood pressure change, and gross neuropathology to ascertain which parameter might best account for the behavior observed after brain injury. The results showed that decreasing neurologic scores correlated well with increasing atmospheres of injury (Pearson's r 0.71, p < 0.001) but not with the rise in systolic blood pressure caused by the trauma (n = 29). Coma scores did correlate with the cross sectional area of ponto-mesencephalic lesions (Pearson's r = 0.51, p < 0.01) and proved to be significantly different in animals grouped according to lesion size of less or more than 3 mm in length (t test, p < 0.01). Thus the CCS and the pharmacologic properties of propofol permit an early analysis of the neurologic status in the feline fluid percussion model of brain injury. These procedures could facilitate the evaluation of early biochemical changes that affect behavior and of therapies designed to ameliorate the deleterious effects of head injury. PMID- 8411217 TI - Unilateral cortical contusion injury in the rat: vascular disruption and temporal development of cortical necrosis. AB - Cerebrovascular disruption and cortical pathology resulting from either moderate (M-TBI) or severe (S-TBI) traumatic brain injury produced by a pneumatically driven cortical contusion device were assessed in adult male rats sacrificed at 6 and 24 h or 8 and 30 days after injury to the right sensorimotor cortex. Epidural, subdural, subarachnoid, petechial (cortex and corpus callosum), and/or intraventricular hemorrhage was present in all animals, more extensively and severely following S-TBI. At 6 or 24 h after TBI, acidophilic (acid fuchsin positive) neurons were numerous and widespread (S-TBI > M-TBI) in the ipsilateral contused cortex. By 8 days few acidophilic neurons were present in peri-impact regions of the ipsilateral neocortex, and none were detected in cortex 30 days postinjury. Both M-TBI and S-TBI groups had enlarged ipsilateral cortical volumes (edema) at 6 and 24 h post-contusion. Eight and 30 days after injury the mean volume of cortical necrosis was significantly larger in S-TBI than in M-TBI rats, and cortical necrosis in both TBI conditions increased between 8 to 30 days postinjury. These results indicate that this pneumatically-driven contusion device produces reliable and consistent primary and secondary cortical histopathology, the extent of which is related to the severity of initial injury. PMID- 8411218 TI - Selective loss of neurons from the thalamic reticular nucleus following severe human head injury. AB - The GABAergic neurons of the thalamic reticular nucleus, or nucleus reticularis thalami (RT), have been implicated as important components in attentional processing systems. Neurons in the RT are exquisitely sensitive to degeneration following kainic and domoic acid toxicity, experimental global ischemia, human cardiac arrest, and experimental closed head injury in nonhuman primates. The present study was performed to establish whether the selective loss of human RT neurons occurred following severe head injury. Brains from 37 human nonsurvivors of head injury were examined for evidence of RT neuronal loss. RT lesions in were found in 36 of 37 cases, representing 65 of 73 (89%) of the reticular nuclei examined. The incidence of RT lesions was similar in all age groups: 13 of 14 cases (92.9%) in the pediatric (< or = 16 years) group, 33 of 37 (89.2%) in the young adult (18-45 years) group, and 19 of 22 (86.4%) in the older adult (> 45 years) group. RT lesions were characterized by loss of one fourth to three fourths of the neurons from the region of the nucleus associated with the frontal cortex and thalamic mediodorsal (MD) and ventrolateral (VL) nuclei. Sparing of RT neurons correlated highly with the presence of extensive frontal cortical lesions, suggesting that an intact corticothalamic projection was necessary for RT degeneration following head injury. A pathologic cascade with a prominent excitotoxic component is proposed. The loss of these inhibitory thalamic reticular neurons and the resultant thalamic and neocortical neuronal dysfunctions may underlie some forms of attentional deficits that persist following head injury. PMID- 8411219 TI - Sensory evoked potentials for selective monitoring of the rat spinal cord: a cerebellar evoked potential to assess ventral cord integrity. AB - The purpose of this study was to evaluate two types of ascending sensory evoked potentials (SEPs) in the rat and their capacity for selective monitoring of dorsal versus ventral spinal cord integrity. SEPs were elicited by direct sciatic nerve stimulation. A cerebellar evoked response was recorded over the paramedian lobule of the cerebellar hemisphere (CEPpml) while somatosensory evoked potentials (SSEPs) were simultaneously recorded over the sensorimotor cortex. All components of the CEPpml and SSEP except the longest latency positive waves were present in each animal. At stimulus intensities of 3 to 10 mA, no significant changes in latency of the peaks were observed, but amplitudes of the longer latency responses tended to increase throughout this stimulation range. Unilateral sciatic stimulation resulted in bilateral cortical responses, larger ipsilaterally for the CEPpml, and contralaterally for the SSEP. Selective spinal cord lesions demonstrated N9 and P14 of the CEPpml to be mediated primarily through the ventral spinal cord, while P14 and N19 of the SSEP were conducted primarily through the dorsal columns. Sectioning of the cerebellar peduncles abolished N9 and P14 of the CEPpml despite persistence of the SSEP. This study demonstrates that selective assessment of the ventral and dorsal spinal cord is possible in the rat by monitoring SEPs. PMID- 8411220 TI - Effects of nimodipine on posttraumatic spinal cord ischemia in baboons. AB - Posttraumatic ischemia appears to be largely responsible for the extension of lesions in acute injury of the spinal cord. In the present study, we have evaluated the putative improvement of axonal function by the calcium channel blocker nimodipine after acute trauma of the spinal cord. Three techniques were used: (1) spinal cord blood flow (SCBF) using a scanographic technique with stable xenon, (2) somatosensory evoked potentials (SEPs), and (3) magnetic resonance imaging (MRI). Thirteen baboons were used in this study. Acute trauma was achieved by compression of the spinal cord at level L1 by applying pressure for 5 sec with an inflated balloon catheter injected with Ringer's solution. Following the injury, one group (n = 5) received a saline infusion (placebo) for seven days, and a second group (n = 8) received a nimodipine infusion (0.04 mg/kg/h) during the same period of time. SCBF and SEP were first recorded prior to trauma. SCBF, SEPs, and MRI were then recorded on the day of the injury and eight days prior to histologic examination of the spinal cord. In these studies nimodipine significantly improved SCBF. The decrease in SCBF observed at day one and day eight following trauma was significantly reduced in the treated group. Two baboons in the treated group also showed improvement of axonal function as assessed by SEP. No significant difference was observed with MRI, however, histologic study revealed that the lesions were significantly smaller in the treated group. Based on these observations we conclude that a week of nimodipine treatment following spinal cord injury enhances SCBF, limits the size of the spinal cord lesion, and perhaps improves functional recovery. PMID- 8411221 TI - Comparison of single and combination drug treatment strategies in experimental brain trauma. AB - Effects of single-drug and combination drug treatments were examined in a model of lateral fluid percussion-induced traumatic brain injury (TBI) in rats. Treatments included the opioid receptor antagonist nalmefene, the thyrotropin releasing hormone (TRH) analog YM14673, the noncompetitive N-methyl-D-aspartate (NMDA) antagonist dextrorphan, nalmefene + dextrorphan, YM14673 + nalmefene, YM14673 + dextrorphan, and equal volume saline. Single-dose treatment with nalmefene, YM14673, or dextrorphan at 30 min after trauma each significantly improved behavioral recovery at two weeks as compared with vehicle-treated controls, confirming earlier studies with these agents. No combination treatment was superior to treatment with the most effective individual drug alone. Combination treatment with the TRH analog and the NMDA antagonist resulted in significantly less effectiveness than treatment with either drug alone. These findings indicate the need for preclinical studies to examine potential drug-drug interactions in the treatment of central nervous system (CNS) trauma. PMID- 8411222 TI - Is dietary fat a risk factor for prostate cancer? PMID- 8411223 TI - Drug designers target Ras for cancer treatment. PMID- 8411224 TI - RAC ponders its purpose: to be or not to be. PMID- 8411225 TI - More gene therapies for AIDS slated to begin. PMID- 8411227 TI - Clinical trial reimbursement reform sought. PMID- 8411226 TI - Doctors debate different breast biopsy techniques. PMID- 8411228 TI - Ubiquitous heat shock proteins deal with stress, cancer. PMID- 8411229 TI - The importance of p53 gene alterations in human cancer: is there more than circumstantial evidence? PMID- 8411230 TI - Biological and clinical implications of heat shock protein 27,000 (Hsp27): a review. AB - Heat shock and other environmental and pathophysiologic stresses stimulate synthesis of heat shock proteins (Hsps). These proteins enable the cell to survive and recover from stressful conditions by as yet uncompletely understood mechanisms. Hsp27 is an important small Hsp (molecular weight, 27,000) found in human cells--both cancer cells and normal cells. This protein, besides its putative role in thermotolerance, is of special clinical interest because of recent data suggesting it may also play a role in drug resistance. In adults, Hsp27 is found particularly in several cell types such as breast, uterus, cervix, placenta, skin, and platelets. Although low-molecular-weight (small) Hsps have been found to be involved in embryogenesis of Xenopus and Drosophila, they have not been detected in human fetal organs. Regulation of expression of the Hsp gene (also known as HSPB1) has been considered a paradigm of gene regulation and is actively being studied in both prokaryotes and eukaryotes. In prokaryotes, the major Hsp genes are transcriptionally regulated by positively and negatively acting transcription factors. In eukaryotes, the genes encoding Hsps contain a regulatory DNA motif (inverted repeats of the pentameric sequence nGAAn) known as the heat shock element. Hsp27 may function as a molecular chaperone and in signal transduction pathways of different cell regulators, and Hsp27 and other Hsps may be active in development of resistance to stressful conditions and agents including cytotoxic drugs. Study findings indicate that some but not all estrogen positive breast cancers express Hsp27, and overexpression of Hsp27 has been associated with both good and poor prognosis. In endometrial carcinomas, the presence of Hsp27 is correlated with the degree of tumor differentiation as well as with the presence of estrogen and progesterone receptors. Studies suggest, however, that detection of Hsp27 should not be considered to be a method for identifying hormone-responsive tumors or detecting estrogen receptors. Hsp27 seems to be a biochemical marker of estrogenic endometrial response. In patients with cervical cancer, Hsp27 is predominantly expressed in well-differentiated and moderately differentiated squamous cell carcinomas. In addition, expression of Hsp27 seems to be a negative prognostic factor for gastric cancer. Different isoforms of Hsp27 have been found in lymphoid tissue of patients with acute lymphoblastic leukemia, and the protein has also been associated with viral infections. These aspects are summarized and discussed in the present review. PMID- 8411231 TI - Cancer treatment and age: patient perspectives. AB - BACKGROUND: Despite some evidence that age does not meaningfully influence the efficacy or toxicity of cancer treatment, older patients tend to receive less comprehensive cancer therapies. PURPOSE: We conducted a population-based study to evaluate the selection of cancer treatment among the elderly. METHODS: Between September 1 and November 30, 1990, we interviewed by telephone a sample of 628 female Wisconsin residents recently diagnosed with breast (507) or colorectal (121) cancer. The women, aged 20-74 at the time of diagnosis, were identified through Wisconsin's statewide tumor registry. The approximately 30-minute long telephone interview, part of a larger study of cancer etiology, included questions on treatment history, physician specialty, and reasons for the selection of specific therapies. Analyses compared the proportion of subjects with various treatment characteristics according to age (< 65 and > or = 65 years). In evaluating the effect of age on selected therapies, we adjusted summary proportions for stage of disease using the indirect method. The Mantel Haenszel chi square statistic was used to evaluate statistical significance of the differences in proportions. RESULTS: After adjusting for stage of disease at diagnosis, substantial variation was observed in cancer treatment according to age for both breast and colorectal cancer. Older women (> or = 65 years) with breast cancer were less likely than younger women (< 65 years) to have received conservative surgery, radiation, and adjuvant therapy. Older women were, in fact, more likely than younger women to accept mastectomy (P = .03). Consultation with a medical or radiation oncologist was less common among older than younger patients (57% versus 73%). Older women were also less likely to have alternative therapies presented to them (19% versus 31%). While older patients were less likely to have been offered adjuvant treatments, like chemotherapy (P < .01), they were also more likely than younger women to reject these treatments when offered (P = .01). These differences were observed in both breast and colorectal cancer patients. Regardless of age, the most common reasons for not selecting treatments were physicians' recommendations and the desire for more comprehensive treatment. Concern about side effects, however, was more frequently reported by older women (P = .07). CONCLUSION AND IMPLICATION: Patients' ages influence the choice of treatment. Physicians offer older women with cancer different treatments from those offered to younger women and are less likely to recommend specialist consultation. Physicians' advice and description of toxicity may influence patients' selection of treatment. However, older patients' concerns about the consequences of cancer treatment may also influence treatment choice. PMID- 8411232 TI - Improving consistency in cervical cytology reporting. AB - BACKGROUND: During the 1970s, the Papanicolaou method of classifying cervical cytology specimens and reporting diagnoses was replaced by more descriptive reporting systems. The plethora of reporting terms caused much confusion and a lack of standardization. To improve this situation, "The Bethesda System for Reporting Cervical/Vaginal Cytologic Diagnoses" was approved at a National Cancer Institute Workshop in 1988. In Australia, the Victorian Cervical Cytology Registry (VCCR) was established in 1989. Because of the absence of a standard format for reporting cervical cytology in that country, a coding schedule was developed by local cytopathologists. While the pattern of reporting smear diagnoses was found to be reasonably consistent within individual laboratories, substantial variation in reporting abnormal cervical smear diagnoses by 29 laboratories in Victoria, Australia, was observed. In 1992, a working party of the National Health and Medical Research Council of Australia proposed that a modified Bethesda System be adopted by Australian laboratories. PURPOSE: The aim of this study was to promote more uniform reporting of cervical/vaginal cytologic diagnoses by cytopathology laboratories in Victoria, Australia. METHODS: From the computer database, VCCR staff identified 80 slides that had been registered during the first half of 1991 and that covered the range of low-grade reports and negative reports. Each slide was identified by research number only. Two sets of 40 slides were compiled. Of the 29 laboratories that had worked with the VCCR during 1991, 22 agreed to participate in this study in 1992. One slide set was sent to each laboratory. An evaluation of the intralaboratory and interlaboratory consistency in reporting a set of 40 slides was undertaken. Analysis of the results compared the degree of consistency using current descriptive terminology that operates locally in Victoria with that which would pertain if the proposed Australian modification to the Bethesda System were adopted. RESULTS: Intralaboratory agreement with previously reported slides was low on the squamous descriptor (49% agreement with original report) but higher on the human papillomavirus descriptor (76% agreement with original report) when the results were analyzed using the current terminology. Wide variation in reporting was apparent between laboratories; only 5% of the slides had agreement by all laboratories. Both intralaboratory and interlaboratory agreement improved substantially when results were grouped into the categories of the proposed Australian modification of the Bethesda Reporting System. CONCLUSION AND IMPLICATION: Substantial improvement in the consistency of reporting cervical cytology specimens would be likely if terminology incorporating the broad categories of the Bethesda System were adopted. PMID- 8411233 TI - Expression of basic fibroblast growth factor in primary human renal tumors: correlation with poor survival. PMID- 8411235 TI - Road-deicing salt and cancer: the need for further study. PMID- 8411234 TI - Severe fluorouracil toxicity in a patient with dihydropyrimidine dehydrogenase deficiency. PMID- 8411236 TI - Early cancer clinical trials: safety, numbers, and consent. PMID- 8411237 TI - Screening for breast cancer: what should national health policy be? PMID- 8411238 TI - NCI proposes new breast cancer screening guidelines. PMID- 8411239 TI - NCI's proposed breast cancer screening guidelines. PMID- 8411240 TI - Cancer program praised and criticized as plans are forged for the 21st century. PMID- 8411241 TI - Peer review put to the test: credibility at stake. PMID- 8411242 TI - Prostate Cancer Prevention Trial launched. PMID- 8411243 TI - Statistical and ethical issues in the design and conduct of phase I and II clinical trials of new anticancer agents. PMID- 8411244 TI - Cancer mortality among New Mexico's Hispanics, American Indians, and non-Hispanic Whites, 1958-1987. AB - BACKGROUND: Racial and ethnic differences in cancer incidence and mortality are well documented. New Mexico's ethnically and racially diverse population provides an opportunity to further examine ethnic and racial differences in cancer occurrence. PURPOSE: To address differences in cancer mortality among the state's Hispanics, American Indians, and non-Hispanic Whites, we examined mortality data collected from 1958 through 1987. METHODS: Sex and age-specific and age-adjusted cancer mortality rates were calculated for all sites and specific sites for American Indians, Hispanics, and non-Hispanic Whites. From 1958 through 1987, deaths due to malignant neoplasms were coded according to the International Classification of Diseases. The categories of malignant neoplasms investigated were chosen, in part, to minimize bias due to changes in disease classification. Ethnicity was assigned by the Bureau of Vital Statistics on the basis of information on death certificates. Denominators were derived from the censuses of 1960, 1970, 1980, and 1990. Age-standardized mortality rates were calculated for 5-year periods (1958-1962, 1963-1967, 1968-1972, 1973-1977, 1978-1982, and 1983 1987), with the 1970 U.S. population as the standard. We also examined age specific rates by time period. RESULTS: Within each of New Mexico's ethnic groups, overall cancer mortality increased over the 30-year time span, and the cancer mortality rates were greater for males than for females. For most major cancer sites, mortality rates for New Mexico's non-Hispanic Whites were comparable with data for U.S. Whites. American Indians had the lowest rates for most sites, whereas cancer mortality rates for most sites among Hispanics were intermediate between the two other groups. However, Hispanics and American Indians had higher mortality rates for cancers of the gallbladder, cervix, and stomach compared with non-Hispanic Whites throughout most of the study period. Several other cancer sites showed major mortality rate differences among these racial and ethnic groups, including cancers of the colon, rectum, breast, bladder, lung, ovary, and uterus. We also observed strong temporal trends of increasing or decreasing mortality rates for several cancer sites. CONCLUSIONS: Race and ethnicity have been strong determinants of cancer mortality in New Mexico. Within the span of one generation, cancer mortality has changed substantially for some cancer sites in each of the population groups studied. IMPLICATIONS: These mortality data underscore the need for appropriately designed etiologic studies of cancer in diverse racial and ethnic groups. Such etiologic studies could provide new insights concerning risk factors for cancer and useful data for developing race- and ethnic-specific cancer control strategies. PMID- 8411245 TI - Radiation-induced breast cancer: long-term follow-up of radiation therapy for benign breast disease. AB - BACKGROUND: From the 1920s through the 1950s, radiation therapy was used in Sweden as a treatment for benign breast diseases. It is now known that exposure of the breast to ionizing radiation increases the relative risk of subsequent breast cancer, especially for younger women. However, the degree to which the patient's age contributes to the elevation of risk for subsequent development of breast cancer is not yet completely understood. PURPOSE: The purpose was to study the risk of breast cancer after irradiation of the female breast and, in particular, to analyze the duration of the effect and the risk for women older than 40 years at first exposure. METHODS: In this cohort study, data were obtained through population-based registers. The exposed group consisted of 1216 women (median age, 40 years) who, during the period spanning 1925 through 1954, received radiation therapy for benign breast disease. The reference group consisted of 1874 women (median age, 36 years) who had the same diagnosis during that time period but did not receive radiation therapy. The radiation doses were determined from the original medical records (mean dose, 5.8 Gy; range, 0.003 50.1 Gy). The follow-up lasted up to 60 years after first exposure. The incidence rate ratio was analyzed with Poisson regression models. RESULTS: The total number of breast cancers in the exposed cohort was 198 versus 101 in the unexposed cohort. Overall, the radiation-associated incidence rate ratio was 3.58 (95% confidence interval = 2.77-4.63). The dose-response gradient was statistically significant (P < .001) but leveled off at higher doses. The incidence rate ratios decreased starting about 25 years after first exposure but were at increased levels throughout the entire follow-up period. The incidence rate ratio decreased with age at first exposure but was significantly increased (P < .001) even when the age at time of first exposure was greater than 40 years. CONCLUSIONS: Total dose, age at first exposure, and time since first exposure were all determinants of the incidence rate ratio of breast cancer after exposure of the breast to ionizing radiation. A statistically significant increase in the incidence of breast cancer following radiation treatment of various benign breast diseases was observed even among women older than 40 years at the time of first treatment. IMPLICATIONS: These findings need to be considered when weighing the relative benefits versus risks of generalized screening of younger women for breast cancer by mammography. PMID- 8411246 TI - Are esterases involved in multidrug resistance? PMID- 8411247 TI - Re: Relationship between environmental tobacco smoke exposure and carcinogen hemoglobin adduct levels in nonsmokers. PMID- 8411248 TI - Re: Blood levels of organochlorine residues and risk of breast cancer. PMID- 8411249 TI - Re: Lung cancer incidence among patients with beryllium disease. PMID- 8411250 TI - Toward a model for early stages of human breast carcinogenesis. PMID- 8411252 TI - Hyperthermia research rekindled after poor U.S. start. PMID- 8411251 TI - Nighttime light studied as possible breast cancer risk. PMID- 8411253 TI - Heidelberg center works on viruses, genes, and a swifter computer. PMID- 8411254 TI - Error sparks scrutiny of cervical cancer screening. PMID- 8411255 TI - Smoking control effort moves to second phase. PMID- 8411256 TI - Xenograft model of progressive human proliferative breast disease. AB - BACKGROUND: Progression of proliferative breast disease has been associated with increased risk for development of invasive carcinoma. Cell lines have been developed to facilitate the study of this process. Human cell line MCF10A originated from spontaneous immortalization of breast epithelial cells obtained from a patient with fibrocystic disease, and cell lines MCF10AneoN and MCF10AneoT were created by stable transfection of these cells with the neomycin-resistance gene and either the HRAS gene or the mutated T-24 HRAS gene, respectively. PURPOSE: Our goal was to develop an experimental model of progressive human proliferative breast disease. METHODS: MCF10A, MCF10AneoN, and MCF10AneoT cells were injected subcutaneously into the dorsal flank of male nude/beige (C57/BALB/c nu/nu bg/bg) mice (12 mice for each cell type). These mice were examined periodically for formation and persistence or growth of palpable nodules. One mouse per group was killed 1 week after cell injection; thereafter, mice were observed as long as possible. Cells were recovered from palpable lesions by enzymatic dissociation of the excised lesions. Cells re-established in tissue culture from a week-14 tumor (MCF10AneoT.TG1) were injected into 12 male nude/beige mice. Southern blot hybridization analysis of the HRAS gene locus and cytogenetic analyses were performed. RESULTS: Transplanted MCF10A and MCF10AneoN cells formed transient, small palpable nodules that regressed and disappeared during the 4th and 5th weeks. In 10 of the 12 mice, T-24 HRAS gene-transfected MCF10A cells (MCF10AneoT) formed small, flat nodules that persisted for at least 1 year. Three of these xenografts became carcinomas. One (removed 7 weeks after transplantation) was an undifferentiated carcinoma composed of polygonal cells with large, vesicular nuclei and numerous mitoses. The second (removed after 14 weeks) was an invasive squamous cell carcinoma. The third (removed after 56 weeks) was a moderately differentiated adenocarcinoma. Initially, xenografts of MCF10AneoT.TG1 cells showed intraductal proliferative changes; after 23 weeks, the lesions showed histologic features resembling those seen in atypical hyperplasia of the human breast, and later lesions showed characteristics of carcinoma in situ. The MCF10 lineage of cells of three MCF10AneoT.TG1 xenografts was confirmed by DNA fingerprinting and karyotype analysis. CONCLUSIONS: MCF10AneoT and MCF10AneoT.TG1 comprise a transplantable xenograft model that produces a broad spectrum of human proliferative breast disease. IMPLICATIONS: The reproducible establishment of representative stages in early breast cancer progression from the MCF10 model offers a new opportunity to analyze critical events of carcinogenesis and progression in breast cancer. PMID- 8411257 TI - Importance of multiagent chemotherapy regimens in ovarian carcinoma: dose intensity analysis. AB - BACKGROUND: In the previous meta-analysis of dose intensity (dosage) of chemotherapy in advanced ovarian cancer, we analyzed data on cyclophosphamide, altretamine (hexamethylmelamine), doxorubicin, and cisplatin. Only cisplatin showed statistically significant association of complete and partial clinical response with dose intensity. PURPOSE: This analysis updates the previous results and further characterizes response to cisplatin alone or in multiagent regimens. METHODS: We analyzed data from 18 regimens containing platinum (cisplatin or carboplatin) that were used in nine new randomized trials, in addition to data from the 60 groups of patients in our previous study in which responses were reported. Relative dose intensity was calculated as a fraction of the dosage of a drug in the standard regimen of cyclophosphamide, altretamine, doxorubicin, and platinum (CHAP). We performed single and multiple regression analyses to determine the relationship between disease outcome and relative dose intensity for cyclophosphamide, platinum, and doxorubicin alone or in combination. RESULTS: The association between outcome and dose intensity for platinum alone or in multiagent regimens was statistically significant. This association was of borderline significance for cyclophosphamide alone but was not significant for this drug in multiagent regimens. There were insufficient data to test the relationship for doxorubicin as a single agent, but in multiagent regimens, the relationship was borderline (P = .05). Multiagent regimens containing platinum produced greater response rates than platinum alone for any fixed, planned relative dose intensity for platinum. CONCLUSIONS: Our results support other published findings that use of cyclophosphamide and doxorubicin increases the efficacy of single-agent platinum. Relative dose intensity values for cyclophosphamide used alone were larger than those used in multiagent regimens, which might explain why the relationship between relative dose intensity and outcome for cyclophosphamide was not significant for use in multiagent regimens. Similarly, none of the multiagent regimens incorporated doxorubicin at a relative dose intensity for which the drug is found to be effective as a single agent. IMPLICATIONS: Prospective clinical trials are required to test the effect of higher relative dose intensity for doxorubicin and cyclophosphamide added to platinum in advanced ovarian cancer. An important element in the design of prospective trials will be to test for the relative importance of dose intensity versus total dose. This testing is best achieved in a three-arm study design such as that reported in adjuvant treatment of stage II breast cancer conducted by the Cancer and Leukemia Group B. PMID- 8411258 TI - Effects of dietary omega-3 fatty acids on human breast cancer growth and metastases in nude mice. AB - BACKGROUND: Diets rich in omega-6 polyunsaturated fatty acids stimulate the growth and metastases of transplantable mammary carcinomas in rodents, whereas fish oil-containing diets, rich in omega-3 fatty acids, suppress the growth of these mammary tumor cells. PURPOSE: This study was performed to evaluate the effect of a diet rich in menhaden fish oil on the growth and metastases of MDA-MB 435 human breast cancer cells in a mouse model system. METHODS: Ninety female athymic nude mice (Ncr-nu/nu) were fed a 23% (wt/wt) corn oil, omega-6 fatty acid rich diet; after 7 days, 1 x 10(6) estrogen-independent MDA-MB-435 human breast cancer cells were injected into a thoracic mammary fat pad. The 23% corn oil diet was continued for a further 7 days, after which the mice were assigned randomly to one of three diets containing a total of 23% fat, but different proportions of corn oil and menhaden oil (diet Cm: 18% corn oil and 5% menhaden oil, diet CM: 11.5% corn oil and 11.5% menhaden oil, and diet cM: 5% corn oil and 18% menhaden oil). Animal body weights and the surface area of the mammary fat pad tumors were recorded weekly. The mice were killed after 12 weeks on the experimental diets. Primary tumor surface areas and body weights were compared by unpaired Student's t tests, the incidence of lung metastases by the chi-square test, and differences in the total volumes of lung metastases by the nonparametric Mann-Whitney U test. RESULTS: Tumor growth rates in the mice of the group fed diet Cm were significantly greater than for mice of either of the two groups fed diets containing higher levels of menhaden oil. Of the mice with primary tumors, the incidence of macroscopic lung metastases was greater in those fed diet Cm, compared with those fed diet cM (57.7% versus 22.2%; P < .01), but not significantly different from the mice fed diet CM. When metastases did occur, their extent was significantly greater in mice fed diet Cm, compared with those fed diet cM (P < .001). CONCLUSION: These results indicate that a high-fat diet rich in omega-3 fatty acids can suppress human breast cancer cell growth and metastases in this mouse model system. IMPLICATION: Dietary intervention trials to reduce recurrence risk in the postsurgical breast cancer patient should take account not only of the level of fat consumed, but also its fatty acid composition. PMID- 8411259 TI - Elevation of multiple serum markers in patients with stage I ovarian cancer. AB - BACKGROUND: The high overall mortality from ovarian cancer (> 60%) relates, in part, to delays in diagnosis. When ovarian cancer is detected in stage I (International Federation of Gynecology and Obstetrics staging), up to 90% of patients can be cured. Transvaginal sonography can detect early-stage disease with great sensitivity, but it is expensive and lacks specificity. Although serum marker assays could provide a less expensive and more convenient initial screening test, the sensitivity of assays varies. Measurement of serum CA 125 in conjunction with ultrasound screening as a second-line test confers high specificity but detects only about one half of early stage ovarian carcinomas. PURPOSE: The purpose of this retrospective study was to determine whether assays of multiple serum markers would improve sensitivity by detecting a higher percentage of stage I ovarian cancers than the CA 125 assay alone. METHODS: Using immunoradiometric assays, we measured preoperative serum levels of CA 125 tumor associated antigen, macrophage colony-stimulating factor (M-CSF), and OVX1 in 46 patients with stage I ovarian cancer of different histologies and 237 patients with benign pelvic masses. We also assayed sera from 204 apparently healthy women who had participated in a screening trial and remained free from cancer at 1 year of followup. All specimens were obtained from cryopreserved aliquots. Marker levels were considered to be elevated when levels of CA 125 were greater than 30 U/mL, M-CSF levels were greater than 3.1 ng/mL, or OVX1 levels were greater than 12.1 U/mL. RESULTS: At least one of the serum markers was elevated in 98% of patients with stage I ovarian cancer; CA 125 levels were elevated in 67%. By the same criteria, 11% of healthy individuals and 51% of patients with benign pelvic masses had at least one elevated marker value. Thus, the sensitivity of the combination of assays for the three serum markers was significantly greater than the sensitivity of the CA 125 assay (P < .0005) and specificity was moderate. CONCLUSION: A panel of these three tumor markers can identify early-stage ovarian cancer with extremely high sensitivity and moderate specificity. IMPLICATIONS: Elevation of one or more serum markers should be evaluated further as an indication for transvaginal sonography in apparently healthy women. Such a strategy might substantially reduce the expense and improve the specificity of screening compared to the use of ultrasound alone. Prospective studies with a large cohort of patients at high risk for ovarian cancer will be required to confirm these findings. PMID- 8411260 TI - Association of MMP-2 activation potential with metastatic progression in human breast cancer cell lines independent of MMP-2 production. AB - BACKGROUND: Expression of matrix metalloproteinase-2 (MMP-2), the 72-kd type IV collagenase/gelatinase, by cancer cells has been implicated in metastasis through cancer cell invasion of basement membranes mediated by degradation of collagen IV. However, the abundance of this latent proenzyme in normal tissues and fluids suggests that MMP-2 proenzyme utilization is limited by its physiological activation rather than expression alone. We previously reported activation of this proenzyme by normal and malignant fibroblastoid cells cultured on collagen I (vitrogen) gels. PURPOSE: Our purposes in this study were 1) to determine whether MMP-2 activation is restricted to the more invasive human breast cancer cell lines and 2) to localize the activating mechanism. METHODS: Zymography was used to monitor MMP-2 activation through detection of latent MMP-2 (72 kd) and mature species of smaller molecular weight (59 or 62 kd). Human breast cancer cell lines cultured on plastic, vitrogen, and other matrices were thus screened for MMP-2 activation. Collagen I-cultured cells were exposed to cycloheximide, a protein synthesis inhibitor, or to protease inhibitors to determine the nature of the MMP 2-activating mechanism. Triton X-114 (TX-114) detergent extracts from cells cultured on collagen I or plastic were incubated with latent MMP-2 and analyzed by zymography to localize the MMP-2 activator. RESULTS: MMP-2 activation was only induced by collagen I culture in the more aggressive, highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines (Hs578T, MDA MB-436, BT549, MDA-MB-231, MDA-MB-435, MCF-7 ADR) and was independent of MMP-2 production. MMP-2 activation was detected in cells cultured on collagen I gels but not in those cultured on gelatin gels, Matrigel, or thin layers of collagen I or IV, gelatin, or fibronectin. Collagen-induced activation was specific for the enzyme species MMP-2, since MMP-9, the 92-kd type IV collagenase/gelatinase, was not activatable under similar conditions. MMP-2 activation was inhibited by cycloheximide and was sensitive to a metalloproteinase inhibitor but not to aspartyl, serine, or cysteinyl protease inhibitors. MMP-2 activation was detected in the hydrophobic, plasma membrane-enriched, TX-114 extracts from invasive collagen I-cultured cells. CONCLUSION: Collagen I-induced MMP-2 activation is restricted to highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines, is independent of MMP-2 production, and is associated with metastatic potential. Our findings are consistent with plasma membrane localization of the activator. IMPLICATIONS: The MMP-2 activation mechanism may represent a new target for diagnosis, prognosis, and treatment of human breast cancer. PMID- 8411261 TI - Poor prognosis of p53 gene mutation and nuclear overexpression of p53 protein in inflammatory breast carcinoma. PMID- 8411262 TI - NBSS revisited--again. Miller comment. PMID- 8411263 TI - Re: Dietary fat and breast cancer. PMID- 8411264 TI - NTera 2 cells: a human cell line which displays characteristics expected of a human committed neuronal progenitor cell. AB - We have identified a human cell line with a phenotype resembling committed CNS neuronal precursor cells. NTera 2/cl.D1 (NT2/D1) cells expressed nestin and vimentin, intermediate filament (IF) proteins expressed in neuroepithelial precursor cells, as well as MAP1b, a microtubule-associated protein (MAP) expressed in human neuroepithelium. NT2/D1 cells also expressed the cell adhesion molecules NCAM and N-cadherin which are thought to be important in cell-cell interactions within the neuroepithelium. These NT2/D1 cells also expressed small amounts of NF-L, alpha-internexin, NF-M, and MAP2c, indicating that they are committed to a neuronal fate. Previous studies have shown that, following RA treatment, a proportion of NT2/D1 cells terminally differentiate into neurons and that this occurs via an asymmetric stem cell mode of differentiation. In light of the identification of the neuroepithelial phenotype of NT2/D1 cells we decided to examine more closely the relationship of in vitro neurogenesis in NT2/D1 cells, during RA treatment to that of neurons in vivo. Three days after RA treatment, islands of NT2/D1 cells showed increased expression of neurofilament proteins and increased phosphorylation of NF-M. By 10-14 days, these cells began to resemble neurons morphologically, i.e., with rounded cell bodies and processes. These neuronal cells were clustered into clumps which rested on top of a layer of progenitor cells. In this upper layer, the neurons began to express MAP2b and tau and extinguished their expression of nestin. Recently, we developed a method for obtaining pure cultures of neurons from RA treated NT2/D1 cells. The phenotype of these postmitotic neurons is clearly dissociated from that of the untreated NT2/D1 cells. Given the data obtained in this study and the characterization of the neurons derived from NT2/D1 cells, we propose that NT2/D1 cells are a committed human neuronal precursor cell line which retains some stem cell characteristics and is capable only of terminal differentiation into neurons. PMID- 8411265 TI - B-50 (GAP-43) in the spinal cord caudal to hemisection: indication for lack of intraspinal sprouting in dorsal root axons. AB - Sprouting of dorsal root axons has been suggested to occur in the mature cat spinal cord caudal to a hemisection at a low thoracic level sparing the dorsal columns. The lesion interrupts supraspinal descending projections, while leaving ascending collaterals of dorsal root axons intact. This hypothesis was re evaluated by comparing the light and electron microscopic immunoreactivity of B 50 (GAP-43) on both sides of the postulated target regions for sprouting, the intermediate gray and the dorsal horn. The neural-specific phosphoprotein B-50 is involved in regenerative and developmental axonal outgrowth and synaptic plasticity. The light microscopic distribution pattern and density of B-50 immunostaining, measured by quantitative densitometry, were bilaterally symmetrical in all segments below the hemisection 3.5, 8, 14, 21, and 56 days postoperatively, as they were in the intact animal. Ultrastructurally, growth cone-like profiles were not detectable during putative periods of sprouting in regions of interest. After removal of degenerated axon terminals, vacated postsynaptic places appeared to be covered by astrocytic processes. These results indicate that, under the present experimental conditions, sprouting of primary afferents in adult cats is unlikely to be involved in functional plasticity after removal of descending pathways. PMID- 8411266 TI - Dye-induced photolesion in the mammalian retina: glial and neuronal reactions. AB - Irradiation in the presence of a dye applied to the extracellular space is known to produce degenerative features in irradiated neurones and fibers. In the present study, we confirmed the potential use of this procedure as a lesion technique by showing the removal of degenerating elements as part of the glial reaction to the lesion. The dye Rose Bengal was applied to the vitreous body of a rat eye and a T-shaped irradiation pattern was projected onto the retina within the absorption band of the dye. Degenerative features were restricted to the irradiated area, which could be readily identified from its shape. Retinae examined after various survival times showed that macrophages invaded the damaged area within 1 day, and that mitotic activity of reactive glial cells subsequently occurred in the vicinity of the wound. Both cell types were identified by their structural features. Macrophages were also revealed by a staining technique using the dye Nile Red, whereas reactive glial cells were immunolabeled with an antibody directed against the glial fibrillary acidic protein. Reactive glial cells helped the macrophages to gradually remove injured cells and damaged processes. Their main task, however, appeared to be in scar formation, since their processes seemed to suture the lips of the wound together and restore the limiting membrane at the inner retina. After 2 months' survival time, the parent ganglion cells of most disrupted axon bundles had retrogradely degenerated, but regenerating ganglion cell axons were also observed. These results provide some new data about healing processes in the retina. They demonstrate that the dye induced photolesion technique can be used to either remove or axotomize selected neurones in neural networks which have been made optically accessible. PMID- 8411267 TI - mRNAs coding for neurotransmitter receptors in rabbit and rat visual areas. AB - Levels of mRNAs encoding neurotransmitter receptors in the visual cortex, lateral geniculate nucleus, and superior colliculus of the rabbit and rat, and properties of the receptors expressed, were studied using Xenopus laevis oocytes. mRNA extracted from these areas was injected into the oocytes, which then acquired functional receptors. Electrical recordings of neurotransmitter-induced membrane currents reflect the relative amounts of mRNAs encoding the corresponding receptors. Receptors to gamma aminobutyric acid (GABA), kainate, glutamate, and serotonin exhibited uniformly high levels of expression, whereas expression of receptors to glycine and N-methyl-D-aspartate was uniformly low. In contrast, the expression of receptors to acetylcholine and substance P was highly non-uniform. Expression of acetylcholine receptors was high in oocytes injected with mRNA from the visual cortex, low for the lateral geniculate nucleus, and very low or absent for the superior colliculus. Conversely, the currents elicited by substance P were large in oocytes injected with superior colliculus mRNA, but were small or absent in oocytes injected with mRNAs from the other regions. Immunohistochemical analysis, at the light and electron microscopic levels, was used to localize choline acetyltransferase, the acetylcholine-synthesizing enzyme, and substance P containing synaptic boutons in the three visual areas. Their presence closely paralleled the potency of mRNAs coding for acetylcholine and substance P receptors. The ability of rat mRNA, from each visual area, to induce neurotransmitter receptors was similar to that observed in the corresponding rabbit mRNAs. In addition to the marked differential distribution of mRNA encoding neurotransmitter receptors in the visual system, our findings reveal the probable existence of as yet uncharacterized receptors, whose new molecular forms may be revealed by further study. Our results also provide the basic information required for subsequent studies on the effect of monocular deprivation on the expression of neurotransmitter receptors in the visual system. PMID- 8411268 TI - Age-related loss of knee joint afferents in mice. AB - Previous work in our laboratory revealed markedly different rates of age-related death of four monoaminergic neuronal populations in the C57BL/6 mouse. Although dorsal root ganglion neurons (DRGns) have been reported not to suffer similar age related death in rodents, we determined if there is age-related death of the subpopulation of DRGns innervating the knee joints of C57BL/6 mice, which are known to develop degenerative arthritis with aging. The somata of dorsal root ganglion neurons innervating the mouse knee joint (KJ-DRGns) were identified by retrograde tracing with Fluoro-Gold (FG). Lumbar ganglia were serially sectioned and the numbers of FG-labelled KJ-DRGns counted at five ages encompassing the animal's life span. Changes in size of the total population of lumbar DRGns (L DRGns) were estimated by counting nucleated somata from every fifth toluidine blue-stained serial section from the L3 and L4 lumbar ganglia at three different ages. Using a computer-assisted video morphometric technique somal areas were measured from random sections to determine the distribution of sizes of neurons in the KJ-DRGn and general lumbar DRGn populations at different ages. Counts of FG-labelled joint afferents were 238.5 +/- 80.3 (mean +/- SD) KJ-DRGns per knee at 2 months of age, declining to 103.2 +/- 20.1 by 24 months, representing a 57% loss over the average life span of the C57 mice. The loss occurred in two phases, with a rapid rate over the first 8 months of life and a more moderate rate of loss over the remaining months. L-DRGn numbers revealed a slower overall rate of loss in comparison to the KJ-DRGn population with an average 33.7% loss over the life span of this mouse. Somal size measurements revealed that the larger sizes of KJ-DRGns were lost over the first 8 months of life, with little change in the distribution of somal sizes thereafter. The distributions of sizes of the L-DRGn population did not change significantly over the life spans of the mice. The data provides evidence that the age-related loss of KJ-DRGns is significantly greater than DRGns in general, and may be particularly apparent in the population of larger sized presumed mechanoreceptor neurons. The loss of the KJ-DRGns is approximately reciprocal to the incidence rate of knee joint osteoarthritis reported for the C57BL/6 mice. PMID- 8411269 TI - Amphiphilic and hydrophilic forms of acetyl- and butyrylcholinesterase in human brain. AB - Human brain acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were sequentially extracted, first with a Tris-saline buffer (S1) and then with 1% (w/v) Triton X-100 (S2). About 20 and 30% of the AChE and BuChE activities were recovered in S1 and most of the remaining enzymes in S2. Main molecular forms of about 10.5 S and 12.0 S, G4 forms of AChE and BuChE, and smaller amounts of 4.5 S and 5.5 S forms, G1 species of AChE and BuChE, were measured in S1. Application of Triton X-114 phase partitioning and affinity chromatography on phenyl-agarose to S1 revealed that 25% of the AChE and none of the BuChE molecules displayed amphiphilic properties. Analysis of the enzyme activity retained by the phenyl agarose showed that G1 AChE constituted the bulk of the amphiphilic molecules released without detergent. Main G4 forms of AChE and BuChE were found in the S2 extract. Eighty and 45% of the AChE and BuChE activities in S2 were measured in the detergent-rich phase by Triton X-114 phase partitioning. Thus, most of the AChE and about half of the BuChE molecules in S2 displayed amphiphilic properties. The main peak of BuChE, a 12.0 S form in gradients made with Triton X 100, splits into two peaks of 9.5 S and 12.5 S in Brij 96-containing gradients. This suggests that hydrophilic G4 BuChE forms are predominant in S1 and that hydrophilic and amphiphilic isoforms coexist in S2. PMID- 8411270 TI - Injury severity grading in trauma patients: a simplified technique based upon ICD 9 coding. AB - The purpose of this study was to develop a simplified method of stratifying patient risk of death based on ICD-9 codes. METHODS: Data were obtained from a statewide trauma registry. A mortality risk ratio (MRR) was derived from a "training" subset by calculating a mortality rate for each ICD-9 code of interest. The independent variables of interest included TS, ISS, and MRRs (for the 1st & 2nd Dx, 1st op, & E code). RESULTS: (n = 37,100). When the 1st Dx and ISS were used as candidate variables in stepwise multivariate modeling, the MRR for the 1st Dx was the first variable to be entered into the model (1st Dx partial R2 = 0.37, ISS partial R2 = 0.02). CONCLUSION: This study shows that the 1st Dx is a better predictor of outcome than ISS. Since ICD-9 codes are more easily obtained and are better predictors of outcome, this study suggests that they may supersede the use of the ISS in injury severity scoring. PMID- 8411271 TI - Lung injury from gut ischemia: insensitivity to portal blood flow diversion. AB - Gut ischemia/reperfusion (I/R) appears to produce pulmonary vascular injury through endotoxin release and cytokine activation. The ability of hepatic reticuloendothelial cells to clear bacterial products may also be impaired during I/R. To test this, diversion of the splanchnic blood flow from the liver into the systemic circulation was performed via a microsurgical portacaval transposition in anesthetized Sprague-Dawley rats (275-375 g). Shunted animals underwent portacaval transposition and were allowed to recover for 7-10 days; sham animals underwent exploration but no shunt was created. The I/R animals were subjected to 60 minutes of reperfusion. All shunts were patent at autopsy. Pulmonary vascular permeability was assessed by measuring tissue retention of Evans blue dye. Gut I/R produced significant increases in pulmonary vascular permeability (46.2% +/- 11.0% vs. 16.4% +/- 3.8% [I/R vs. control]; p < 0.05) regardless of the presence of hepatic bypass (32.7% +/- 9.0% vs. 10.0% +/- 1.4% [I/R vs. control]; p < 0.05). These data indicate that a mediator or mediators of gut origin are responsible for pulmonary vascular permeability changes following gut I/R and are not appreciably modulated by the liver. PMID- 8411272 TI - Diagnosing pneumonia in mechanically ventilated trauma patients: endotracheal aspirate versus bronchoalveolar lavage. AB - We prospectively investigated the diagnostic value of semiquantitative (semiQC) and quantitative (QC) cultures of endotracheal aspirate (ETA) compared with QC of bronchoalveolar lavage (BAL) fluids in 18 mechanically ventilated trauma patients with clinical signs of pneumonia. The general agreement between QC of ETA and BAL was 89% when conventional cutoffs for the QC were used and 94% if the cutoffs were adjusted for previous antibiotic therapy. In all six patients whose clinical diagnoses of pneumonia were considered definite, both QC of ETA and QC of BAL were positive; however, standard semiQC of ETA showed comparable results in this group. On the contrary, in the 12 patients whose clinical diagnoses were uncertain, QC of BAL and ETA were negative in ten patients and in five (50%) of these, pneumonia was eventually excluded. Semiquantitative cultures of ETA were positive in all these patients. Five (28%) patients experienced a decrease in PaO2/FiO2 (> 15% of previous value) 2 hours after BAL, and in three (17%) this derangement persisted for 24 hours. These data suggest that BAL may be hazardous in mechanically ventilated trauma patients and that its use should be restricted to patients in whom the diagnosis is in doubt. PMID- 8411273 TI - Trauma outcomes in the rural developing world: comparison with an urban level I trauma center. AB - Trauma is well known as a major cause of death and disability in the developed world, but has been inadequately studied in developing nations. We reviewed 539 trauma patients admitted from 1987 through 1991 to a rural African hospital, the Holy Family Hospital (HFH) in Berekum, Ghana, and compared these results with 14,270 patients admitted during the same period to a level I trauma center, the Harborview Medical Center (HMC) in Seattle, Washington. At HFH, 59% of patients were seen > 24 hours after injury, compared with 4% of HMC patients (p < 0.001). Only 25% of HFH patients received prehospital care, compared with 82% of HMC patients (p < 0.001). Mean ISS was higher at HMC (10.0 +/- 6.3) than a HFH (6.7 +/- 6.5) (p < 0.001), but trauma mortality rates were identical (6%) at both institutions. Neurologic injuries were the leading cause of death at both HFH (62%) and HMC (54%). There was no significant difference between institutions in mortality for patients with ISS 1-8 (HMC: 0.7% n = 6390; HFH: 0.3%, n = 342). There was a marked decrease in mortality for patients with ISS 9-24 at HMC (3%, n = 3709) compared with HFH (10%, n = 146) (p < 0.001). There was a less pronounced decrease in mortality for patients with ISS > 24 at HMC (41%, n = 1520) compared with HFH (73%, n = 26) (p < 0.01). The type and the severity of injuries causing fatalities in this developing nation suggest that no inexpensive hospital-based changes would improve outcomes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411274 TI - A risk analysis of pulmonary complications following major trauma. AB - Varying institutional definitions and degrees of surveillance limit awareness of the true incidence of posttraumatic pulmonary complications. Prospective review with standardized definitions of 25 categories of pulmonary complications was applied to a university level I trauma service over 3 years to establish the true incidence. Potential injury-related predictors of individual complications were determined using multiple logistic regression analysis and adjusted odds ratios were calculated, thereby controlling for the effect of other covariants. Significance was attributed to p < 0.05. Of 3289 patients meeting MTOS criteria, pulmonary complications occurred in 368 (11.2%). Pulmonary complications account for one third of all disease complications. Significant associations with pneumonia included age, the presence of shock on admission, significant head injury, and surgery to the head and chest. Significant risk for atelectasis occurred in patients with blunt injury mechanism, ISS > 16, shock on admission, and severe head injury. Risks for development of respiratory failure included age > 55 years, the mechanism of "pedestrian struck", and the presence of significant head injury. Risk factors for ARDS included surgery to the head and a Trauma Score < 13 on arrival. Significant predictors for pulmonary embolism included ISS > 16, shock on admission, and extremity and pelvis injuries. The true incidence of pulmonary complications is established with this kind of analysis and focuses attention on (1) groups at high risk for developing complications, (2) groups for which current therapeutic modalities are still ineffective, and (3) defining the need to refocus on prospective research rather than ineffective processes of care. PMID- 8411275 TI - Urban firearm deaths: a five-year perspective. AB - Firearm violence is an ever-increasing element in the lives of the U.S. urban population. This study examined the trends in firearm violence and victims during a 5-year period in the city of Philadelphia. Medical Examiner records of all deaths in Philadelphia County in 1985 and 1990 were reviewed. Demographic, autopsy, and criminal record information was analyzed. There were 145 firearm homicide victims in 1985 versus 324 in 1990, a 123% increase. This was primarily because of deaths among young (age 15-24 years), black male victims. Handguns were involved in at least 90% of firearm homicides in both study years. The use of semiautomatic handguns increased from 24% to 39% during the study period. In 1985, 42% of revolver homicides died at the scene, versus 18% in 1990. However, 5% of victims of semiautomatic weapons fire died at the scene in 1985 versus 34% in 1990. The decrease in survival of semiautomatic weapon victims occurred despite the implementation of six trauma centers within the county, and probably reflects a shift toward high-velocity, high-caliber ammunition. Antemortem drug use and criminal history was common. A total of 54% of victims were intoxicated in 1985 and 61% were in 1990. Cocaine became the most common intoxicant in 1990, with 39% of victims using it during the antemortem period. The percentage of victims with a criminal record increased from 44% to 67%. Although the duration of criminal history decreased from 14 to 6 years, the number of patients with previous drug offenses increased from 33% to 84%..(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411276 TI - Effect of preinjury illness on trauma patient survival outcome. AB - Data from 11,156 patients treated at the four Major Trauma Outcome Study controlled sites were used to estimate the effect on survival of each APACHE II preinjury illness condition (PIC). A case-control methodology was applied; 544 patients (4.8%) had one or more PICs. For each patient with a specific PIC we identified a set of matching patients with no PICs. A patient matches a PIC patient if both have the same mechanism of injury, the same coding of Revised Trauma Score variables (Glascow Coma Scale score, systolic blood pressure, respiratory rate), the same coded age per A Severity Characterization of Trauma) (ASCOT), and if they differ by no more than 0.5 for A, B, and C (the ASCOT components for serious injuries). The estimated survival probability for a PIC patient is either the survival rate for the patient's matched set or, if there are no matches, the patient's ASCOT survival probability. The survival probabilities were used to compare the actual and predicted numbers of survivors for each PIC, using z and W statistics. Computations of z and W were also made using ASCOT survival probabilities for each PIC patient. The results indicate profound effects of five PICs (hepatic, cardiovascular, respiratory, renal, diabetes) on trauma patient outcomes. CONCLUSION: Pre-existing organ dysfunction has a profound effect on patient outcome even after controlling for age, anatomic and physiologic severity, and mechanism of injury. But, because of their relatively low incidence in this sample, PICs did not strongly influence institutional outcome performance as measured by z and W values. PMID- 8411277 TI - Continuous quality improvement applied to a scientific assembly: the history of the Eastern Association for the Surgery of Trauma. AB - INTRODUCTION: The purpose of this study was to measure the quality of the research presented at the Eastern Association for the Surgery of Trauma (EAST) for the first 5 years. METHODS: Abstracts from the scientific assemblies of EAST, 1988-1992, were reviewed. Five elements were identified for each abstract, the research question, the research design, the sampling method, the validity of the conclusions, and if the research question was answered. These were identified because of their significant impact on the quality of the research presented in the abstract format. RESULTS: Two hundred two abstracts were reviewed. There was a significant improvement in research design, in the sampling method, in the validity of the conclusions, and in the proportion of research questions answered. There was an initial improvement in the presence of a research question, but it was not sustained. CONCLUSION: The results clearly demonstrate an improvement in the quality of the abstracts chosen for presentation at the scientific assembly of the Eastern Association for the Surgery of Trauma. Further improvements can be made by adding the category of 'research question' or 'purpose' and requiring a structured abstract. PMID- 8411278 TI - On the nature of things still going bang in the night: an analysis of residency training in trauma. AB - In the 1982 Presidential Address to the Society of University Surgeons, Trunkey reported on the inadequacy of surgical education in trauma care. His conclusions were based on American Board of Surgery operative experience data compiled from residents completing surgical training in 1980. The purpose of this study was to compare current resident operative experience in trauma surgery with the American Board of Surgery data from 1980. Yearly resident operative experience data obtained from the Residency Review Committee from 1987 through 1991 were analyzed. The relationship between the percentile rank and the number of operative cases was defined using linear regression. The percentile rank of residents performing a specified number of operative cases was computed using a linear regression coefficient. The results were then compared with previously published 1980 American Board of Surgery summary data. Resident operative experience in trauma surgery was stable over the 5-year period investigated and no significant trends were identified. Comparison of the data from 1980 to 1991 revealed that the percentage of residents performing less than ten cases decreased markedly, from 18% to 9%. Moreover, the percentage of residents claiming fewer than 50 cases declined from 86% to 29%. Based on this analysis, it appears that resident operative experience dramatically increased from 1980 to 1987 and has since remained stable. The reasons for this are unclear but undoubtedly involve the accuracy of reporting operative experience, Residency Review Committee operative trauma definitions, and the actual number of trauma surgery cases available for trainees.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411279 TI - Resident supervision in the operating room: does this impact on outcome? AB - Resident supervision by faculty is a sine qua non of surgical education, yet objective standards for supervision are difficult to quantify. Over a 12-month period, using departmental data on morbidity, mortality outcome, and faculty status in the operating room, the association between complications, death, and attending physician presence were analyzed by Chi-square tests of association in 2 x 2 contingency tables, or by the Mantel-Haenszel Chi-square to control for a stratifying variable. A total of 4417 cases were reported. Attending physicians were either scrubbed or present in the OR 91.8% of the time, although there was considerable variation among services. The overall mortality rate was 6.2% and complications occurred in 7.0% overall. Greater attending physician presence was significantly associated with lower mortality and complication rates overall. When stratified by service, the association was less marked. However, presence of attending physicians varied significantly by service. To adjust for this variation, elective services were compared with all the "nonelective" services. When this categorization was used as the stratifying variable, the association between increased attending physician involvement and decreased complication and mortality rates was statistically significant (Mantel-Haenszel Chi-square, p < 0.0005 for both). PMID- 8411280 TI - Characteristics of trauma centers and trauma surgeons. AB - We surveyed directors of trauma at 408 trauma centers (as indicated by the state chairmen of the American College of Surgeons' Committee on Trauma [ACSCOT]). Of the 408 trauma directors 290 (71%) responded with information relative to their hospitals and the 1537 general surgeons taking trauma call. Altogether, 75% of the surgeons worked on an identified trauma service, 80% belonged to a cadre of surgeons identified as expert, 52% were viewed as full time, 25% provided in house staff coverage, and 76% had completed an ATLS course. Six percent of the entire group were 60 to 73 years old and demonstrated a commitment to trauma equal to that of their younger cohorts. As a group, the older surgeons took less call, but when community or the institutional call needs were identified, the older surgeons met the challenge. PMID- 8411281 TI - Early management of civilian gunshot wounds to the face. AB - We analyzed 54 civilian patients (1988-1992) with gunshot wounds (GSWs) of the face to review the management principles and results. Urgent airway control was needed in 18 cases (33%): by orotracheal intubation in 13, cricothyroidotomy in two, tracheostomy in two, and nasotracheal intubation in one. Central nervous system injury was seen in 12 (22%): 40% with orbital, 38% with mid-face, and 0% with lower face injuries. Two patients died of intracranial injuries (mortality, 4%). Vascular injury was present in five patients (9%), all detected by angiography. The local complication rate was 39% in the presence of intra-oral injury and 0% without intra-oral injury (p < 0.001). The maxilla was the most commonly fractured facial bone (41%) followed by the mandible in 28%. All maxillary, orbital, and zygomatic fractures were treated without reduction. One of the seven nasal fractures required open reduction for deformity. Six of the 15 mandible fractures were treated without reduction. Of eight patients treated with closed reduction, one developed nonunion. One patient treated with immediate open reduction developed osteomyelitis of the mandible and nonunion. Five patients (9%) had palate injuries. Two of them later developed intraoral fistulas following conservative treatment. The airway needs immediate attention in GSWs of the face. Computed tomographic scanning of the head or spine should be done when the bullet trajectory is above the lower face (the level of the mandible). Angiography is indicated when the trajectory of the bullet is suggestive. A conservative approach that effectively reduces the fractures is the procedure of choice. Open reductions should not be performed in the initial treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411282 TI - Hemodynamic responses to penetrating spinal cord injuries. AB - Although the hemodynamic response to blunt spinal cord injury has been well described, much less is known about the responses to penetrating spinal cord injuries. In order to elucidate any differences, we reviewed the last 75 patients treated over the past 12 years with penetrating spinal cord injuries. There were 67 men and eight women; the mean age was 26.2 years (range, 15-59 years); 73 patients suffered 120 gunshot wounds; one patient was injured with an ice pick; one was stabbed twice. The offending missile causing spinal cord injury entered the neck in 24%, the thorax in 56%, and the abdomen in 20%. Nine patients (12%) were complete quadriplegics and 49 patients (65%) were complete paraplegics; 69 patients (92%) had no rectal tone; 17 patients (22%) had incomplete injuries. Despite the high proportion of complete spinal injury (78%), only 18 patients (24%) were hypotensive in the field. Five additional patients became hypotensive in the ED. Of the 23 patients with hypotension, 18 (74%) had significant blood loss to explain their low blood pressure. The mean HR was 100 beats/minute in the field (range, 50-130 beats/minute) and 90 beats/minute in the ED. Only five patients (7%) demonstrated the classic presentation of neurogenic shock (hypotension and bradycardia). This classic presentation of neurogenic shock is rare following penetrating spinal cord injury. Despite evidence of a complete spinal cord injury on initial physical examination, hypotension is usually secondary to blood loss in these patients. A careful search for sources of blood loss is mandatory before ascribing hypotension to spinal injury. PMID- 8411283 TI - Lactate clearance and survival following injury. AB - Previous reports cite optimization of O2 delivery (DO2) to 660 mL/min/m2, O2 consumption (VO2) to 170 mL/min/m2, and cardiac index (CI) of 4.5 L/min as predicting survival. We prospectively evaluated 76 consecutive patients with multiple trauma admitted directly to the ICU from the operating room or emergency department. Patients had serum lactate levels and oxygen transport measured on ICU admission and at 8, 16, 24, 36, and 48 hours. Patients were analyzed with respect to survival (S) versus nonsurvival (NS), lactate clearance to normal (< or = 2 mmol/L) by 24 and 48 hours, hemodynamic optimization as defined above, as well as Injury Severity Score (ISS), ICU stay (LOS), and admission blood pressure. All patients achieved non-flow-dependent VO2. There was no difference in CI, DO2, VO2, or ISS when S was compared with NS. All 27 patients whose lactate level normalized in 24 hours survived. If lactate levels cleared to normal between 24 and 48 hours, the survival rate was 75%. Only 3 of the 22 patients who did not clear their lactate level to normal by 48 hours survived. Ten of the 25 nonsurvivors (40%) achieved the above arbitrary optimization criteria. Fifteen of the survivors never achieved any of these criteria. Optimization alone does not predict survival. However, the time needed to normalize serum lactate levels is an important prognostic factor for survival in severely injured patients. PMID- 8411284 TI - Nitric oxide production is inhibited in trauma patients. AB - Elevated levels of nitrates/nitrites, the stable endproducts of nitric oxide (NO), were recently observed in septic patients. In this setting, NO maintains blood flow by vasodilation and inhibition of platelet aggregation. Trauma patients were found to have low plasma levels of nitrates/nitrites, even when they developed sepsis. The current study substantiated that trauma patients have suppressed production of NO; reductions in plasma nitrate/nitrite levels correlated with low urinary excretion of these endproducts. Nitric oxide production was upregulated in trauma patients with clinical infection compared with trauma patients without infection, but was still significantly suppressed compared with nitric oxide production in normal controls. The inability of trauma patients to produce NO may be an important component of the susceptibility of these patients to infection. PMID- 8411285 TI - Antibody synthesis to peptidoglycan polysaccharide after ischemic injury of the intestine. AB - BALB/c mice (ischemia: 31; controls: 15) were studied to investigate the effects of intestinal ischemia on antibody synthesis to peptidoglycan polysaccharide (PGPS), a ubiquitous bacterial antigen found in both gram-positive and gram negative bacteria. The gut ischemia model was produced by placing a vessel loop around the superior mesenteric vessels for 45 minutes. All animals in the ischemia group had visible gut ischemia. Eighteen animals (58%) in the ischemia group survived to 24 hours and all experienced total recovery of gut viability. Single-cell suspensions of splenic lymphocytes were made. After 5 days of culture with lipopolysaccharide, anti-PGPS immunoglobulin concentrations in culture supernatants were measured by ELISA using high-titer BALB/c anti-PGPS serum as control. The synthesis of immunoglobulin by 10(5) lymphocytes was significantly increased in the ischemia group compared with the controls. These results represent the translocation of bacteria after intestinal ischemia, and this antibody response may be important in resistance to sepsis and multiple organ dysfunction attributed to bacterial translocation. PMID- 8411286 TI - Growth hormone attenuates Na(+)-dependent hepatic amino acid transport in endotoxemic rats. AB - The effects of human growth hormone (GH) on hepatic Na(+)-dependent amino acid transport were studied in endotoxin-treated rats. Adult rats received GH (6 mg/kg BW subQ q 12 hours x 4 doses) or vehicle prior to a single dose of Escherichia coli endotoxin (LPS, 7.5 mg/kg BW IP). Four hours after LPS administration, livers were excised and hepatocyte plasma membrane vesicles (HPMVs) were prepared by differential and Percoll density gradient centrifugation. Hepatocyte plasma membrane vesicle transport of [3H]-MeAIB, a highly selective system A substrate, [3H]-glutamine, a selective system N substrate, and [35S]-cysteine, a system ASC substrate, were measured by a rapid mixing/filtration technique. Vesicle purity and functionality were assessed by marker enzyme measurements and classic overshoots and timecourses that showed similar vesicle size. Endotoxin treatment resulted in a two-fold increase in the activities of systems N and ASC, which was the result of an increase in carrier Vmax (Km was unchanged), and a four-fold stimulation of system A. Pre-treatment with GH diminished the endotoxin-induced increase in Na(+)-dependent amino acid transport by 60%-80%; this reduction in carrier-mediated transport activity was a result of a decrease in Vmax, consistent with a decrease in the number of functional transporter proteins in the plasma membrane. Growth hormone treatment attenuates the endotoxin-induced increase in the activities of the major Na(+)-dependent transporters in rat liver. This may diminish hepatic ureagenesis and spare amino acids for peripheral protein synthesis and thereby explain, at least in part, the ability of growth hormone to promote positive nitrogen balance in catabolic states. PMID- 8411287 TI - Hemodynamic effects of carbon dioxide pneumoperitoneum during mechanical ventilation and positive end-expiratory pressure. AB - Laparoscopy is frequently used for diagnosis and treatment of critically ill trauma patients. Its effects on cardiopulmonary performance in the intensive care unit patient population, however, are not well-defined. This study evaluated the effects of positive end-expiratory pressure (PEEP) and carbon dioxide (CO2) pneumoperitoneum on hemodynamic function during mechanical ventilation. Five anesthetized, mechanically ventilated adult swine were monitored with pulmonary artery and arterial catheters at 0, 5, 10, 15, and 20 cm H2O PEEP without, and then with 15 mm Hg CO2 pneumoperitoneum. A hemodynamic profile, analyses of arterial and mixed venous blood gases and mixed venous hemoglobin oxygen saturation values were obtained at each data point. Compared with the non insufflated group, CO2 pneumoperitoneum significantly increased central venous pressure, mean arterial pressure, mean pulmonary artery pressure, pulmonary vascular resistance index, and stroke index for the range of PEEP levels. With PEEP of 10 cm H2O, hemodynamic changes in non-insufflated animals were not statistically significant, but with CO2 pneumoperitoneum, stroke index and left ventricular stroke work index were decreased at 5 cm H2O PEEP, as was cardiac index at 10 cm PEEP. Pulmonary gas exchange was not affected by CO2 pneumoperitoneum. The results indicate that, in this paradigm, CO2 pneumoperitoneum for laparoscopy increases ventricular afterload and exacerbates the adverse effects of PEEP. These findings could be clinically significant in critically ill patients. PMID- 8411288 TI - Use of base deficit to compare resuscitation with lactated Ringer's solution, Haemaccel, whole blood, and diaspirin cross-linked hemoglobin following hemorrhage in rats. AB - Base deficit (BD) has been shown to be a sensitive measure of the degree and duration of inadequate perfusion. We developed a rat model of hemorrhagic shock based on achieving a fixed BD of 13 +/- 1 mmol/L before resuscitation. Using this model, we compared the efficacy of resuscitation with lactated Ringer's solution (LR), Haemaccel (a colloid), and whole blood with that of diaspirin cross-linked hemoglobin (DCLHb, Baxter Healthcare Corp.) by evaluating improvements in BD and restoration of base excess (BE, positive correlate of BD) for 60 minutes following resuscitation. The DCLHb was superior to LR and Haemaccel in restoring and maintaining BE following resuscitation, and was able to restore BE as rapidly as whole blood at half the volume. At 60 minutes, DCLHb at twice the shed blood volume maintained BE at higher (more positive) values compared with all other treatment groups. We conclude that DCLHb is at least as effective as whole blood and superior to LR and Haemaccel in restoring BE within the first 60 minutes following resuscitation in this hemorrhagic shock model. PMID- 8411289 TI - Effect of lesion volume on cerebral hemodynamics after focal brain injury and shock. AB - Cerebral blood flow (CBF) varies unpredictably in patients after head injury and hemorrhagic shock. Proper treatment requires knowledge of ischemic versus hyperemic flow. The degree to which the size or severity of the injury may contribute to CBF abnormalities is unknown. We hypothesized that lesion size is a determinant of postinjury CBF. We measured cerebral and systemic variables in a porcine model of focal cryogenic brain injury and hemorrhagic shock over a 5-hour period. Swine were randomized to receive either a large or small lesion followed by hemorrhage. In the small lesion group traumatic brain injury, followed by shock and resuscitation, produced a significant and sustained elevation in bihemispheric regional CBF and cerebral oxygen delivery that was significantly greater than that observed in either the large lesion group or the controls (p < 0.05). There were no significant differences between the experimental groups in volume of hemorrhage, intracranial pressure, cerebral perfusion pressure, arterial oxygen content, or PaCO2. These data suggest that the volume of injured tissue may determine post-resuscitation CBF, and that interventions to reduce cerebral blood volume (i.e., hyperventilation) may not be universally applicable in all head injured patients. PMID- 8411290 TI - Prophylactic vena cava filter insertion in severely injured trauma patients: indications and preliminary results. AB - Pulmonary embolism (PE) remains a significant problem in trauma patients. A 5 year review at this institution revealed 25 PEs (seven fatal) in 2525 admitted trauma patients (1% incidence). Three groups of high-risk patients were identified: (1) those with severe head injury and coma; (2) those with spinal cord injuries with neurologic deficit; and (3) those with pelvic and long bone fractures. The relative risk of PE in these high-risk patients was 21 to 54 times that of the general trauma population. Beginning in July 1991, as prophylaxis against PE, vena cava filters (VCF) were inserted in patients whose injuries placed them in a high-risk group. Thirty-four patients had VCFs inserted percutaneously in the radiology suite without complications. On follow-up examination, 17.6% developed documented lower extremity deep vein thrombosis. There were no PEs. Overall, the incidence of PE in the general trauma population was significantly decreased from 1% to 0.25% (p < 0.05; chi 2). We conclude that insertion of VCFs in high-risk trauma patients is safe and efficacious in decreasing the incidence of PE. PMID- 8411291 TI - The utility of routine daily chest radiography in the surgical intensive care unit. AB - Routine morning chest x-ray films (CXRs) are widely obtained in surgical intensive care unit (SICU) patients. During a 1-month time period we prospectively evaluated 525 routine morning CXRs in patients admitted to the SICU of a university trauma center (n = 256) or a suburban affiliate hospital (n = 269) to assess the impact of these CXRs on patient care. All CXRs were read by radiologists. Data on position of medical devices (CVP lines, endotracheal tubes, etc.) and cardiopulmonary (CP) findings were collected. A total of 1028 medical devices were evaluated. Fifty-five (5.4%) were considered to be in a minor incorrect position that did not adversely affect patient care and only 13 (1.3%) devices required repositioning for patient care or safety. Seventy-eight CXRs were read as normal. There were 775 CP findings on the remaining 477 CXRs. When compared with previous CXRs, only 12% (89 of 775) of the findings were considered new, 65% were unchanged, 14% were improved, and 15% demonstrated worsening of a known finding. Of the 89 new CP findings, only three had any potential clinical impact (pneumothorax in two, effusion in one). These data demonstrate an extremely low yield of clinically significant and unsuspected new CP findings or device malposition on the routine morning CXR. We conclude that routine daily chest radiography should be abandoned and that the need for a morning CXR should be based on clinical necessity. PMID- 8411292 TI - Diagnostic laparoscopy as an adjunct to selective conservative management of solid organ injuries after blunt abdominal trauma. AB - To investigate the efficacy of diagnostic laparoscopy (DL) as an adjunct in patient selection for conservative management of solid organ injuries (SOI) following blunt abdominal trauma, 15 patients with injuries documented by computed tomographic (CT) scanning were prospectively evaluated. Diagnostic laparoscopy was performed in an attempt to characterize SOI, to evaluate the abdomen for associated occult injuries, and to select patients for conservative management or laparotomy. The 15 patients had CT evidence of 17 SOIs (nine spleen, eight liver), and DL allowed adequate visualization of 15 of the 17 injuries. Occult hollow viscus injury was discovered in 2 of 15 patients (one colon, one small bowel) and required laparotomy. In the remaining 13 patients, DL revealed ongoing hemorrhage in four patients and poor visualization in one patient that prompted laparotomy (four splenorrhaphy, one hepatorrhaphy). Conservative management was employed in the treatment of eight patients with findings of minor injury or adequate hemostasis on DL. The average transfusion requirement in this group was 1.8 U. No patient failed conservative management. There were no complications attributable to DL. These data demonstrate that DL may become an effective adjunct in patient selection for conservative management of SOI following blunt abdominal trauma. PMID- 8411293 TI - Common issues for parents in a methadone maintenance group. AB - Change has been noted in some of the group members. For example, one long-term group member who never spoke when she was in a previous group that focused on addiction-related problems has become the role model for other members. In this group, she says, she feels more competent. She is appropriately outspoken and has established relationships with her children which consistently underpin the parent-child boundaries that need to be drawn. One specific issue she resolved involved her 19-year-old son and his girlfriend who was pregnant with the group member's grandchild. By role playing the different positions that each of these people held in the home (the group member was paying the rent and had the right to set the rules, the girlfriend was feeling unaccepted by the group member and was acting out inappropriately, and the son was caught between trying to please the two females), the group member was able to clarify for herself how to approach future conflicts. A second member has improved her relationship with her youngest son (age 3) but still struggles with her 15-year-old daughter. What has been most effective for her was teaching her to count to 10 before responding to what she considers the goading of both of her children. In addition, she was feeling pressured by her mother, in whose house she lives, to have her son toilet trained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411294 TI - Acupuncture heroin detoxification: a single-blind clinical trial. AB - The increasing prevalence of HIV infection among injection drug users mandates the development of innovative treatments. While extensive clinical experience suggests that acupuncture detoxification is both safe and acceptable to those in withdrawal, little research has been conducted to assess its efficacy as a treatment modality. In this first controlled study of acupuncture heroin detoxification, 100 addicted persons were randomly assigned, in a single-blind design, to the standard auricular acupuncture treatment used for addiction or to a "sham" treatment that used points that were geographically close to the standard points. Attrition was high for both groups, but subjects assigned to the standard treatment attended the acupuncture clinic more days and stayed in treatment longer than those assigned to the sham condition. Additionally, attendance varied inversely with self-reports of frequency of drug use, suggesting that those with lighter habits found the treatment modality more helpful. Limitations of the study are discussed. PMID- 8411295 TI - Pharmacotherapeutic management strategies for mentally ill substance abusers. AB - In order to determine psychiatrists' strategies for managing the pharmacotherapy and treatment of mentally ill substance abusers, the authors constructed a series of standardized clinical vignettes, each with five multiple-choice treatment interventions. The vignettes, based on clinical experience, depict difficult dilemmas involving the use of medication in the treatment of a variety of dually diagnosed patients. The authors surveyed a heterogeneous reference group of 112 psychiatrists with these vignettes. The psychometric properties of the resultant Scale for Treatment Integration of the Dually Diagnosed (STIDD) are briefly described. The ten vignettes and the treatment interventions chosen by the reference group are described in detail. The results indicate that most clinicians favor an integrated treatment approach which attempts to address both diagnoses. PMID- 8411296 TI - Compliance with tuberculosis treatment in methadone-maintained patients: behavioral interventions. AB - Tuberculosis has increased dramatically in the United States. Noncompliance with treatment is high. The purpose of this investigation was to achieve compliance with prophylactic TB treatment and simultaneously decrease drug use in a high risk group of intravenous drug users. Two studies were conducted. Study 1: Subjects were 9 chronic opiate users who tested positive for tuberculosis and were placed on isoniazid (INH) and methadone. Methadone was dispensed contingent upon INH ingestion throughout. A within-subject, A-B design with contingency management interventions on drug use was implemented. RESULTS: Compliance with INH was 100% in 8 patients. Cocaine use remained high. Study 2: Two patients, meeting same criteria as Study 1, participated in a within-subject A-B multiple baseline design. Methadone was dispensed contingent upon INH ingestion throughout. Successive decreases in cocaine use were reinforced in the contingent phase. RESULTS: Compliance with INH was high. During contingency, both patients had over 40% cocaine-free urine samples compared with 0% at baseline. This investigation serves as a model for achieving compliance with TB treatment in opiate users. PMID- 8411297 TI - Sex role stereotypes and clinical judgement: how therapists view their alcoholic patients. AB - In the course of a selection procedure for inpatient treatment in an addiction clinic, it was repeatedly noticed that the attitude of staff-members towards male alcoholics was more confronting and critical as compared to a more empathic and supportive attitude towards female alcoholics. Two different processes may account for this phenomenon. Firstly, male and female alcoholics may differ in their interpersonal behaviour towards therapists, and subsequently these different behaviours evoke different attitudes and behaviours in therapists (interaction-hypothesis). Secondly, these different attitudes towards male and female patients may emerge from preconceived ideas among therapists about male and female alcoholics (stereotype-hypothesis). This study describes the stereotypes held by therapists regarding the interpersonal behaviour of male and female alcoholics. Furthermore, it explores the influence of self perceived interpersonal behaviour of therapists on these stereotypes. The results suggest, that the differences in attitude and behaviour towards alcoholics are--at least partly--the result of different male and female alcoholic stereotypes held by therapists. These stereotypes were related to the self perceived interpersonal behaviour of the therapist. Pretreatment matching of patients and therapists should be based on interpersonal attitude rather than on sex. PMID- 8411298 TI - The Beech Hill Hospital/Outward Bound Adolescent Chemical Dependency Treatment Program. AB - Ninety-one adolescents (74 males and 17 females, mean age = 16.5, range = 14-20) admitted to an in-patient treatment facility with a substance use disorder were followed over a 1-year period post-treatment. Follow-up phone interviews were conducted with each patient and a parent at 3-month intervals. Minnesota Multiphasic Personality Inventory (MMPI) and Personal Experience Inventory (PEI) data were collected along with detailed psychosocial assessments to determine what factors predicted successful treatment outcomes. At 1-year post-treatment, 47% reported complete abstinence from alcohol and other drugs. Survival analyses indicated that participation in a self-help program such as Alcoholics Anonymous (AA) and severity of drug use and psychopathology were associated with relapse risk. Patients with severe psychopathology and drug use scores who were not attending AA were 4.5 times more likely to relapse than patients with low scores who attended AA. PMID- 8411299 TI - On pejorative labeling of cocaine exposed children. PMID- 8411300 TI - Within-person variation in urinary sodium, potassium and creatinine concentrations, and their relationship to changes in the blood pressure of adult male Gambians. AB - Variation in urinary electrolyte and creatinine concentrations was studied in untimed, casual specimens obtained from 65 adult Gambian males over a 15-month period during which up to nine specimens were obtained from each subject. Measurements of blood pressure were made at the times when specimens were collected. The within-person variation in urinary creatinine, sodium and potassium concentrations were all found to be larger than that between persons. Within individual subjects, the sodium concentration tended to be comparatively low, and the potassium concentration high, in specimens in which the creatinine concentration was high. This suggests that the excretion of sodium was reduced at times of relative dehydration while the excretion of potassium may have been unrelated to the urine flow. The data also suggest that changes in the state of hydration were associated with changes in blood pressure since an individual subject's pressure was inversely related to the creatinine concentration. The negative relationship of the sodium:potassium ratio with creatinine concentration casts doubt on the simple use of this ratio as an indicator of dietary intake. After adjusting for creatinine concentration by multiple regression, changes in systolic pressure were shown to be positively correlated with changes in sodium concentration. These observations show the importance of replicating measurements of urinary electrolyte concentrations. The demonstration of complex inter relationships between blood pressure and urinary concentrations of creatinine, sodium and potassium emphasizes the need for care in the interpretation of findings from causal, untimed urine specimens. PMID- 8411301 TI - Dapsone poisoning. AB - Two cases of suicidal dapsone ingestion are reported. These patients presented with marked cyanosis and had significant methaemoglobinaemia on admission. One patient also had acute psychosis and haemolytic anaemia. Both patients responded to intravenous methylene blue therapy and nasal oxygen inhalation and recovered without any sequelae. PMID- 8411302 TI - Seroepidemiology of hepatitis B, delta and human immunodeficiency virus infections in Hamadan province, Iran: a population based study. AB - The epidemiologic features of HBV, HDV and HIV in the general population of Hamadan province of Iran were studied. A total of 4930 subjects (1649 males and 3281 females) constituted the study population. Seropositivity for any HBV marker was found in 25.72% of the subjects. Of these, 2.49% were carriers of HBsAg, 18.09% were positive for anti-HBs and 5.13% for anti-HBc alone. HBeAg, anti-HBC IgM, and anti-HDV were present in 13.8, 6.5 and 2.4% of HBSAg carriers, respectively. Antibody to HIV was detected in none of the 4930 sera tested. Prevalence of HBV infection was lowest in children and young adults < 19 years and highest in adults > 60 years. No significant difference was observed between the rates in males and females. Family size and prevalence of HBV infection were unrelated but an inverse relation was found between HBV infection and education. Unmarried men and women showed the lowest (18.26%) and widows and divorcees the highest (51.59%) rate of HBV infection. Our results suggest that horizontal transmission is likely to be the primary mode of acquisition of HBV infection in children and young adults. Also infection is partly transmitted before or soon after birth to babies of HBsAg-carrier mothers. Socioeconomic and demographic variables have a greater impact on the prevalence of HBV infection than blood or medical care variables in our population. PMID- 8411303 TI - The prevalence and classification of epileptic seizures in Nigerians with sickle cell anaemia. AB - A clinical and electroencephalographic study of the prevalence and pattern of epileptic seizures in 96 patients with sickle-cell anaemia (M:F 1:1, median age 12 years, range 2-45 years) attending our hospital is reported. Ten patients (M:F 1:1.5, median age 21 years, range 16-25 years) had epileptic seizures, giving a relatively high prevalence of 10.4%. Of these, one patient had primary generalized tonic-clonic seizures and four had secondarily generalized tonic clonic seizures. Five patients had partial seizures without generalization. Only two patients were on any antiepileptic drug therapy. Our findings suggest that though epilepsy is common in SCA and the seizures seen are liable to poor control and increased risk for mortality, the patient is unlikely to present voluntarily for therapy. Physicians attending to young adults with SCA should specifically attempt to elicit a history of epileptic seizures in their patients. PMID- 8411304 TI - The disease pattern of elderly medical patients in Rwanda, central Africa. AB - In a study of the disease pattern of the elderly in Rwanda, all patients aged 60 or more, hospitalized in a one-year period at the Medical Department, University Hospital, Butare, were examined prospectively. One hundred and ninety-two patients were included; most were subsistence farmers having a mainly vegetarian diet and living in large families. Infections (37.5% of the patients) and liver cirrhosis (31.8%) were the problems most frequently encountered. Primary hepatocellular cancer was diagnosed in 5.7% of the patients and was the most frequent malignancy. The hospitalized elderly occupied 17.5% of the available beds in the Medical Department. Their disease pattern was different from that of younger patients, making heavier demands on the medical resources. Malaria and upper intestinal inflammation were less frequent in the elderly; liver cirrhosis, primary hepatocellular cancer, pneumonia, prostatic cancer, cardiovascular pathology, chronic renal pathology and chronic lung disease were more prevalent. Several age-related conditions frequently observed in industrialized countries (e.g. coronary heart disease, stroke, gallstones, renal cysts, dementia) were rare. The study thus illustrates the concept of 'secondary aging': to the primary changes induced by the aging process, additional alterations are added which depend upon the environment and the lifestyle, resulting in a varying disease pattern. Health policies thus must take into account that the demographic transition in developing countries may result in a pattern of diseases different from that seen in industrialized countries; care must be taken when transposing data obtained from elderly populations in industrialized countries. PMID- 8411305 TI - A serological survey for leptospirosis in the Enugu area of eastern Nigeria among people at occupational risk. AB - A serological examination of sera from 661 human volunteers in various occupations comprising 248 coal miners, 138 butchers and abattoir workers, 213 local farmers and 62 hospital laboratory personnel from various communities in Enugu and environs of eastern Nigeria was undertaken between January 1990 and March 1991. Leptospiral antibody titres of 1:100 and above were present in 89 (13.5%). The highest prevalence of antibodies to individual serovars were canicola 21 (23.6%), hardjo 17 (19.1%), pomona 13 (14.6%), icterohaemorrhagiae 11 (12.4%), pyrogenes 8 (9.0%), autumnalis 8 (9.0%) and grippotyphosa 7 (7.9%). Among the occupational groups examined, the coal miners were particularly at risk with a prevalence rate of 41 (46%), followed by the butchers/abattoir workers 26 (29.2%), farmers 18 (20.2%) and hospital laboratory personnel 4 (4.5%). PMID- 8411306 TI - Acute chloroquine administration increases renal sodium excretion. AB - The effect of a 2-hour intravenous chloroquine infusion (0.015, 0.030 and 1.25 micrograms min-1) on renal fluid and electrolyte handling was investigated in the saline infused, Inactin anaesthetized rat. Blood pressure and glomerular filtration rate were not affected by chloroquine administration, remaining around 128 mmHg and 2.4 ml min-1, respectively throughout the 5-hour post-equilibration period. Chloroquine produced an increase in Na+ and Cl- excretion without affecting the urine flow. By 1 hour after the start of treatment (0.03 micrograms chloroquine min-1) the Na+ excretion rate had increased to 14.5 +/- 2.1 mumol min 1 (n = 6), and was significantly (P < 0.01) greater than in control animals (8.6 +/- 1.0 mumol min-1) at the corresponding time. Parallel but lesser increases in Cl- excretion rates were also observed. The plasma aldosterone and corticosterone levels following either 10, 30 or 120 minutes infusion of chloroquine at 0.03 micrograms min-1 did not differ statistically from each other or from control values. It is concluded that acute chloroquine administration induces an increase in Na+ excretion. The mechanism of this natriuresis cannot be established from the present study, but is likely to involve altered tubular handling of Na+. PMID- 8411307 TI - Clinical filarial disease in two ethnic endemic communities of Orissa, India. AB - To evaluate the possible role of ethnicity in susceptibility to filarial infection, a comparative study of the prevalence of filarial infection was initiated in an endemic village inhabited by two ethnic populations of mainlanders and tribals. An age and sex matched sampled population of 591 mainlanders and 106 tribals was studied by detailed clinical and parasitological (60 mm3 blood) examinations. Sera collected from both population groups (26 each) matched for clinical stage of infection were analysed for humoral immune responses such as antifilarial IgG, circulating filarial antigen and immune complex level. The overall prevalences of clinical disease and infection in both mainlanders (34.18 and 14.4%) and tribals (25.47 and 17.9%) were comparable. However, both annual average adenolymphangitic attack rate (1.77 year-1) and the prevalence of chronic filarial disease (22.6%) amongst tribals were significantly lower. No true elephantiasis was observed in tribals. No significant difference was observed in their humoral immune response, although the antifilarial antibody of IgG class in all stages of filarial infection was lower in tribals than in mainlanders. The results did not reveal any difference in susceptibility to filarial infection in the ethnic groups. The paucity of progressive lesions observed in tribals possibly reflects a difference in the anatomy of lymphatics or genetic or immunoregulatory mechanisms, that needs further study. PMID- 8411308 TI - Bancroftian filariasis in the Kassena Nankana District of the upper east region of Ghana: a preliminary study. AB - A filariasis prevalence survey was conducted in April 1992 in the Southern sector of the Kassena Nankana District which was the site of the Ghana Vitamin A Supplementation morbidity trial. In all, 106 compounds from five different communities were randomly selected from the vitamin A trial database. All resident compound members of 10 years and above were clinically examined and blood was taken for thick films between 2100 and 0200 hours. Haemoglobin levels were also measured. All the blood slides were examined by the investigators in Navrongo. Two reference laboratories examined 10% each of the slides for quality control. In all, 531 people were examined, 247 males and 284 females. The results showed an overall microfilaraemia rate of 41.1% (95% CI 36.9-45.3%). The only species identified was Wuchereria bancrofti. The most important clinical manifestation was hydrocele, 30.8% of males (95% CI 25-36.6%); followed by elephantiasis of the leg, 3.6% of the study population (95% CI 2.0-5.2%). The mean haemoglobin level of the population was 12.4 g dl-1. There were no significant differences between the communities in clinical or parasitological findings. PMID- 8411309 TI - Acute renal failure in Kuwait--a prospective study. AB - This prospective study was conducted over a period of 18 months (February 1989 to July 1990) in the State of Kuwait. It covered a population of 1,024,211 and eight multidisciplinary hospitals with an in-patient admission of 118,079 per year. Two hundred and twenty-six adult patients with acute renal failure (ARF) were seen and followed up by nephrologists. This made the calculated annual incidence of ARF 14.7 per 100,000 population, nearly five times that reported by the EDTA registry (Biesenbach et al. 1991). Drugs, sepsis and volume depletion were the most frequent causes, with sepsis resulting in 36% cause specific mortality compared to zero mortality with the other two. The overall mortality rate was only 14% which clearly indicated a markedly improved prognosis in cases of ARF. The prognosis in ARF depended on two major factors, viz. the type of aetiological insult and the presence of predisposing associated medical illnesses. Multiple insults, though common, do not affect the mortality rate. Secondary sepsis or gastrointestinal bleeding as a cause of death in ARF was rarely seen in our study. Those who required dialytic support for renal failure had a 45% patient mortality rate in general. Over 40% of our patients were 60 years or older compared with only 3.5% in the local population. This indicated old age as a major risk factor in the development of ARF. The overall mortality in the elderly did not differ from that in the young, but sepsis in the elderly carried a mortality rate of 60% compared to only 14.8% in the younger age group. PMID- 8411310 TI - Recent advances in development of vaccines. PMID- 8411311 TI - Nutritional anthropometry--validity of cut-off points. AB - The present communication seeks to describe an alternate approach of arriving at cut-off levels using ratio of percentiles for (i) differentiating normal children from the undernourished ones, and (ii) identifying the severely undernourished group of children in the community on the basis of body weight and height. It also examines validity of the cut-off levels presently used of weight for age (Gomez and Indian Academy of Paediatrics classification) and height for age parameters. 1. The cut-off levels are independent of the reference standard and, as such, the same cut-off levels could be used irrespective of the standards. 2. It is desirable to have separate cut-off levels for preschool and school age groups. The use of common cut-off points for both the age groups seems to result in loss of sensitivity of the order of over 5 per cent. 3. The use of the 90 per cent cut-off level in the Gomez classification misclassifies some of the normals as undernourished, thereby tending to overestimate the problem of undernutrition in communities. 4. The body weight groups based on the suggested cut-off points do bear a relationship to the prevalence of signs of PEM--the greater the weight deficit, the higher the prevalence of signs of PEM in preschool children. PMID- 8411312 TI - Prevalence of xerophthalmia and efficacy of vitamin A prophylaxis in preventing xerophthalmia co-existing with malnutrition in rural Indian children. AB - Data of 4302 children, 0-6 years old were analysed to study the prevalence of vitamin A deficiency and the efficacy of vitamin A prophylaxis in preventing xerophthalmia co-existing with malnutrition. Manifestation of ocular signs of vitamin A deficiency were seen in 10 per cent children. The prevalence of xerophthalmia was higher in the normal and mild to moderately malnourished children, and lowest in the severely malnourished ones. The ability of vitamin A prophylaxis to curb xerophthalmia was greatest at the extreme ends of the nutritional grade spectrum. These findings have important implications for the existing national Vitamin A Prophylaxis Programme, and suggest that: (a) normal and mild to moderately malnourished children less than 6 years old, should be preferably considered for vitamin A mega dosing; (b) in the management of protein energy malnutrition, vitamin A status of the children should be monitored; and (c) the coverage should be improved as most children are in the mild and moderate degrees of malnutrition. PMID- 8411313 TI - On the molecular interactions between alpha-thalassaemia and sickle cell gene. AB - Using the restriction endonucleases, Bam HI, Bgl II, Hind III and EcoRI, the alpha-gene arrangements were investigated in heterozygotes and homozygotes for the sickle cell haemoglobin (Hb S). In the heterozygotes (Hb AS) group the Hb S level showed a trimodal distribution due to presence of the normal alpha-globin genes (alpha alpha/alpha alpha) or of one (-alpha/alpha alpha) or two (-alpha/ alpha) alpha-genes deletions. The haematological analytes inversely correlated with the associated alpha-thalassaemia (alpha-thal.) genes. In the Hb S homozygotes (Hb SS), associated alpha-thalassaemia was found to ameliorate the clinical manifestations and improved the haematological values. Co-existing triple alpha-gene arrangement, alpha alpha alpha anti 3.7/, with Hb AS did not influence the haematological analytes. In Hb SS, presence of alpha alpha alpha anti 3.7/ resulted in a severe sickle cell anaemia (SCA) with a high severity index (> 11) and with frequent crises, transfusion requirements and hospitalizations. It is suggested that reduced level of alpha-chain ameliorates SCA while excess of alpha-globin chain production gives rise to a severe form of SCA. PMID- 8411314 TI - Patterns of infant weight gain in developing countries. AB - Data relating to weight of infants from 23 developing countries and two western industrialized populations taken from the literature were compared. Growth in the developing countries was extremely variable, but was poor compared to western standards such that by 12 months of age mean weight in all 23 populations was below the NCHS 50th percentile (P < 0.001) and in 11 populations mean weight was below the 10th percentile. Birth weight was not related to growth increment in the first 3 months. Multiple regression analysis showed birth weight and growth increments in the first four 3-month periods post-partum to be highly significantly related to weight at 12 months (P < 0.0001 in all cases). Growth increment in the first 3 months had the greatest effect out of these variables; the effect of the other four variables was similar. These patterns demonstrate the variable impact of environmental factors on growth in the first year of life. PMID- 8411315 TI - HIV, BCG and TB in children: a case control study in Lusaka, Zambia. AB - Even before the onset of the HIV epidemic, studies reported large variations in the protective effect of BCG against TB. The current HIV/AIDS epidemic has increased the incidence of tuberculosis in many countries in sub-Saharan Africa. Thus, local estimates of the effectiveness of BCG are needed which take prevalence of HIV into account for planning strategies for vaccination and TB control programmes. A case control study was conducted in Lusaka, Zambia. The study included 116 TB cases and 154 hospital controls. Eighty-eight per cent of controls had BCG scars compared to 77 per cent of the cases. BCG was not associated with TB in HIV positive children (OR 1.0, 95 per cent CI 0.2, 4.6). However, there was 59 per cent protective effect (OR 0.41, 95 per cent CI 0.18, 0.92) in HIV negative children. The results also suggest an eight times higher risk of TB in HIV positive children. PMID- 8411316 TI - Periventricular haemorrhage in very low birth weight Malaysian neonates. AB - A prospective study was carried out to determine the incidence, clinical presentation, early outcome, and risk factors associated with periventricular haemorrhage (PVH) in 88 (84 per cent) of the 105 consecutive very low birth weight (VLBW) (< 1500 g) Malaysian neonates born in the Maternity Hospital, Kuala Lumpur. Based on the cranial ultrasound findings, PVH was detected in 86 of the 88 neonates (98 per cent, 95 per cent confidence intervals: 95 to 101). Seventeen (20 per cent) of them had grade I, 52 (61 per cent) had grade II, 7 (8 per cent) had grade III and 10 (12 per cent) had grade IV PVH. PVH was detected in all the affected neonates by the fifth day of life. Sixty-four neonates (74 per cent) were symptomatic when PVH was first detected. Shock (P < 0.01), pallor (P = 0.028), low haematocrit of less than 40 per cent (P < 0.01), convulsion (P < 0.001), and bulging of anterior fontanelle (P = 0.019) were significantly more common in the neonates with severe PVH (grades III or IV). Death occurred in 43/86 (50 per cent, 95 per cent confidence interval: 39-61 per cent) of the neonates with PVH before their first discharge from the hospital. Ventriculomegaly developed in 29/43 (67 per cent, 95 per cent confidence intervals: 54.4-81.4) of the survivors with PVH. Our study suggests that PVH is a common problem in the Malaysian VLBW neonates. To reduce the incidence and severity of this condition, prevention of preterm delivery and improvement in the basic facilities for neonatal care would help. PMID- 8411317 TI - Bacteriology of neonatal septicaemia in a rural referral hospital in south India. AB - Out of 640 suspected cases of neonatal septicaemia studied, bacteraemia was detected in 255 (40 per cent) of the infants. Gram negative organisms were predominant (56 per cent) with Pseudomonas, Citrobacter, and Klebsiella as the commonest pathogens. Among the Gram positive organisms both Staphylococcus aureus and Staphylococcus epidermidis were equally prevalent. Staphylococci were mainly responsible for early onset infections, whereas Salmonella typhimurium and Pseudomonas were the main organisms in late onset infections. Group B streptococcal infection was not encountered in this part of the country. PMID- 8411318 TI - Carbohydrate and electrolyte content of some home-available fluids used for oral rehydration in Ghana. AB - Coconut milk and kenkey water (a maize gruel) which are traditionally used for the treatment of diarrhoea in Ghana, were analysed to ascertain their suitability for use in rehydration. The pH, carbohydrate, and electrolyte levels of the food fluids were compared to the recommended UNICEF/WHO ORS to ascertain if these are within physiologically acceptable ranges for the treatment and prevention of dehydration in children with diarrhoea. The carbohydrate and electrolyte levels of kenkey water were found to be comparable to UNICEF/WHO ORS and is suitable for use in rehydration. Coconut milk has the advantages of being fresh, sterile and readily available in most Ghanaian communities. However, use of coconut milk for rehydration cannot be recommended on the basis of its glucose and electrolyte composition. PMID- 8411319 TI - Birth weight patterns in a commercial farming area, rural area and urban area of Karoi District, Zimbabwe. AB - Birth weight patterns were studied in three different socio-economic areas, i.e. commercial farming area (CFA), rural area (RA), and urban area (UA), in a community-based (n = 684) and hospital-based (n = 314) fashion. Mean weight and low birth weight proportions were similar in all three areas, although 17 per cent of all babies were premature in the CFA compared with 7 per cent in the RA (P = 0.005). In the community the mean birth weight was 3.0 +/- 0.1 kg and 2.97 +/- 0.09 kg in hospital. The proportion of newborns weighing less than 2.5 kg was 9 per cent in the community and 15 per cent in hospital (P = 0.006), whereas 18 and 16 per cent were smaller than or equal to 2.5 kg, respectively (P = 0.48). PMID- 8411320 TI - Role of transfusion-mediated viral infections on the lymphocyte subset profile in multi-transfused children. AB - A total of 74 multitransfused (MT) children of beta thalassaemia major were analysed for prevalent viral markers transmitted through transfusion. A higher incidence of serological markers for Hepatitis B virus (HBV), cytomegalovirus (CMV) could be observed in the group of MT children compared to control group. There was a significant trend (chi 2 = 33.4; P < 0.001) in the increase in prevalence of viral markers along with the increase in the number of transfusions. MT children receiving more than 50 transfusions were found to have evidence for at least one or multiple viral infections transmitted through blood. Children receiving more than 50 transfusions were characterized by marked alteration of T3, T8, and B cells while T4/T8 ratio was found to be significantly decreased (P < 0.001) only in the group of children receiving more than 100 transfusions. Relative assessment of the alteration of lymphocyte subsets in various groups of viral infection showed that cases with CMV IgM to have more marked influence on the alteration of T8 cells, T4/T8 ratio, and B cells compared to other groups of viral infections. Reassessment of the lymphocyte subset profile in MT children in the light of CMV IgM positive cases revealed that in children receiving more than 50 transfusions significant alterations of lymphocyte subjects were influenced by the presence of CMV IgM positive cases in these groups. Our study points out that the correlation between the alteration of lymphocyte subset profile and number of transfusion in MT children need to be reassessed in the light of acute CMV infection in the form of CMV IgM. PMID- 8411321 TI - Gender differences and low birth weight with maternal smokeless tobacco use in pregnancy. AB - A preliminary study of maternal smokeless tobacco use, mostly oral applications of burnt tobacco or 'mishri', in pregnancy showed 65 of 178 singleton liveborns occurred to users and 113 to non-users in Bombay, India. Eighty-three newborns, 42 to maternal tobacco users and 41 to non-users were < 2.5 kg birth weight, i.e. low birth weight (LBW; odds ratio 3.2; confidence interval 1.5-6.9; P < 0.001). Stratifying by gender yielded odds ratios of 1.6 (P > 0.1, NS) for male and 6.96 (confidence interval 2.5-19.4, P < 0.0005), for female newborns compared to normal birthweight boys and girls, respectively. Male:female newborns were 80.6:100 in maternal tobacco users compared to 105.5:100 in non-users. Defining LBW as < 2.0 kg yielded an odds ratio of 5.4 (confidence interval 1.8-15.2, P < 0.005) in maternal tobacco users' offspring. For babies weighing 2-2.5 kg at birth it was 2.76 (confidence interval 1.4-5.5, P < 0.01). Maternal use of 'mishri' tobacco in pregnancy may be associated with (1) the offsprings' low birth weight, (2) low birth weights in girls more than in boys; (3) decreased male:female ratio in live newborns, and (4) low birth weight of < 2.0 kg more than of 2-2.5 kg. Studies are needed to substantiate these findings. Gender differences in outcome suggest the in utero effect of maternal smokeless tobacco use on male and female fetuses may differ. PMID- 8411322 TI - Tuberculous spondylitis and intermittent chemotherapy in childhood. AB - The general characteristics and treatment of childhood tuberculous spondylitis are reviewed. Most of the patients (79 per cent) were under 5 years old with girls being in greater number. Twenty-six patients used classical antituberculous chemotherapy and five were given intermittent chemotherapy. There was no significant difference in the mean period of inactivation of the disease (2.8 v. 2.9 months) or in post-treatment angle of kyphosis (33 degrees v. 37.6 degrees) between the classical and intermittent chemotherapy groups. In addition to drug therapy, plaster jacket and immobilization were adequate in four children whereas 27 underwent surgery with only two requiring radical operations. In children with tuberculous spondylitis, intermittent drug treatment is a good alternative due to effectiveness and convenience of use and radical surgery should be reserved for selected cases. PMID- 8411323 TI - Nutrition, immunity and infections: T lymphocyte subpopulations in protein- energy malnutrition. AB - Protein energy malnutrition (PEM) is one of the most frequent causes of secondary immune deficiency states. Alterations either in cellular or humoral immune mechanisms increase the susceptibility to infections in the malnourished organism. Infections aggravate the interrelationship of malnutrition to immune deficiency and infections, resulting in future adverse effects of malnutrition on humoral and cellular immune systems, IgG, IgM, IgA, C3, and T lymphocyte subpopulations were identified in 29 patients with PEM and 15 healthy infants serving as the control group, ranging between 3 and 24 months of age. Patients with PEM demonstrated elevated levels of IgG, IgM and IgA when compared to the control group (P < 0.01, P < 0.01, P < 0.01), C3 levels were significantly lower than the values of the control group (P < 0.01). PMID- 8411324 TI - Long-term prediction of birth weight. AB - On the basis of the hypothesis that undisturbed individual growth in fetal life keeps a constant proportional difference with the standard population 50th percentile, birth weight can be predicted with a single sonographic exploration after the 16th week of pregnancy. Data on 135 singleton pregnancies with accurate dates and delivery at term were used. Sonography was performed between the 17th and 36th weeks of pregnancy, in every case at least 4 weeks before delivery. The observed measurements of BPD, FL, and AC were used for the prediction of their values on the day of delivery, applying the Hadlock equation for the estimation of birth weight. The mean error of birth weight predictions was -1 +/- 11% (SD), with a correlation coefficient between observed and predicted birth weights of 0.62 (P < 0.001). The accuracy was not influenced by the gestational age at the time of exploration. The model underpredicts birth weight of larger fetuses while overpredicting that of lighter ones, but a part of the error could be explained by a change in the growth pattern after modeling. Regardless of the classification considered (i.e., gestational age at sonography or birth weight), over 83% of predictions had an error below 15%. The underlying hypothesis could be useful clinically in modeling and monitoring fetal growth, allowing application of the results of remote sonographic explorations in clinical management. In addition, being able to project fetal weight to the 40th week also is valuable in improving the patient's understanding of fetal growth. PMID- 8411325 TI - Sonographic diagnosis and hemodynamic correlation in veno-occlusive disease of the liver. AB - This study evaluated the role of duplex Doppler sonography in the diagnosis of VOD of the liver after bone marrow transplantation. Sixteen patients with clinical criteria of VOD were studied. The final diagnosis was achieved by transjugular liver biopsy, and the hepatic venous pressure gradient was measured during the procedure to estimate portal pressure. Nine patients (56.2%) had histologically proved VOD. Gallbladder wall thickening was present in 75% of patients with VOD, but in none of those without it (P = 0.01). Gallbladder wall thickening and ascites were present only in patients with portal hypertension. Nine patients (five with VOD and four without it) also were evaluated with duplex Doppler sonography, and no obvious flow abnormalities were detected in the portal vein and hepatic veins. These results suggest that sonography is useful in detecting early signs of portal hypertension pointing to the diagnosis of VOD in patients with bone marrow transplantation. PMID- 8411326 TI - Renal duplex Doppler sonography in asymptomatic women during pregnancy. AB - The purpose of this study was to establish the normal range of the renal RI in pregnant women. Maternal renal RIs were calculated in 61 asymptomatic pregnant patients with at least unilateral pyelocaliectasis. A mean RI was calculated for each kidney. Overall, the mean RI was 0.61 +/- 0.05. The renal RI was > or = 0.70 in 4% (5/121) of kidneys with no statistically significant difference between the mean right (0.62 +/- 0.06) and left (0.60 +/- 0.04) kidney RIs. There was no correlation between trimester of pregnancy or degree of pyelocaliectasis and RI. PMID- 8411327 TI - Adhesive capsulitis of the shoulder joint: usefulness of dynamic sonography. AB - We studied the value of dynamic sonography in adhesive capsulitis of the shoulder joint in patients with clinically suspected frozen shoulder. The sonographic criterion of adhesive capsulitis was continuous limitation of the sliding movement of the supraspinatus tendon against the acromion of the scapula. Arthrography was regarded as a gold standard in the diagnosis of adhesive capsulitis against which the sonographic results were compared. Among 23 patients with arthrographically documented adhesive capsulitis, sonographic examination showed limitation of movement of the supraspinatus tendon in 21. This sonographic sign therefore has a sensitivity of 91%, a specificity of 100%, and an accuracy of 92% for detecting adhesive capsulitis, making dynamic sonography a reliable technique for the diagnosis of this condition. PMID- 8411328 TI - Increased echogenicity in the fetal abdomen: use of DNA analysis to establish a diagnosis of cystic fibrosis. AB - The sonographic finding of increased echogenicity within the fetal abdomen presents a diagnostic dilemma, with a differential diagnosis ranging from normal variation to CF. We report the diagnostic evaluation of four cases, two of which were found to be the result of CF. On the basis of this experience, we believe that persistence of an echogenic bowel pattern, especially with bowel dilation, after 20 weeks' gestation should prompt an evaluation for CF. Using DNA analysis, approximately 75% of the cases involving CF can be detected with noninvasive studies of the parents, and confirmation by amniocentesis is performed only in those cases in which both parents are carriers of known mutations. PMID- 8411329 TI - Operative color Doppler imaging for general surgery. AB - Operative CDI was performed during 125 general surgical operations (53 hepatic, 25 biliary, 24 pancreatic, 7 esophagogastric, 10 pulmonary, 3 kidney transplant, and 3 soft tissue operations). Operative CDI provided beneficial information in 108 of 125 operations (86.4%). On the basis of operative CDI findings, surgical management was altered in 34 of 125 operations (27.2%), most frequently hepatic and pancreatic operations. Operative CDI demonstrated advantages over B-mode imaging in (1) detection and localization of small blood vessels that are impossible or difficult to identify by B-mode imaging, (2) rapid and definitive distinction of blood vessels from other hypoechoic areas, such as tissue spaces and ducts, (3) determination of the relation of tumors to vascular structures such as vascular invasion of carcinoma, (4) confirmation of blood flow to organs after surgical procedures, and (5) clearer needle localization for guidance of needle placement by color motion marking. PMID- 8411330 TI - Galactose-based intravenous sonographic contrast agent: experimental studies. AB - A galactose-based sonographic contrast agent, which produces stable microbubbles capable of traversing the cardiopulmonary circulation, was used to enhance Doppler signals in blood vessels of varying size after intravenous injection. A series of experiments using dogs, rabbits, and woodchucks was conducted to establish the ability of the agent to enhance the reflectivity of normal tissue, tumor tissue, and blood. Although no enhancement was perceptible in tissue on the sonogram, significant enhancement of color and spectral Doppler signals was demonstrated in a variety of vessels. These included the aorta, vena cava, and portal vein as well as such small vessels as those of the retina of the eye, renal cortex, liver parenchyma, and gallbladder wall. Both spectral and color Doppler enhancement was shown in naturally occurring woodchuck hepatomas. Peak Doppler signal enhancement after bolus injection was approximately 10 dB with a dose of 0.01 ml/kg. Recirculation of the agent provided enhancement after intravenous bolus injection for more than 3 min. With a steady intravenous infusion of 0.2 ml/min/kg, Doppler signal enhancement of about 14 dB was maintained continuously for more than 5 min. The results of these animal experiments, in particular in small vessels and with recirculation after intravenous injection, suggest excellent potential for future clinical applications. PMID- 8411331 TI - Sonographic detection of umbilical cord twist. PMID- 8411332 TI - Color Doppler assessment of normal ocular blood flow. AB - The purpose of this study was to determine the role of CDI in the assessment of normal ocular vasculature. Color and pulsed Doppler examinations were performed in 20 healthy subjects. The principal orbital vessels were detected: ophthalmic, central retinal, lacrimal, and posterior ciliary arteries and, in a small percentage, superior ophthalmic, central retinal, and vortex veins; blood flow of arterial vessels was studied by measuring TAV, St, PS, MD, ED, PI, and RI; the morphology of the spectral waveform also was investigated. Our results suggest that duplex color Doppler may be of value in ocular vasculature examination. PMID- 8411333 TI - Perinatal diagnosis of bilateral testicular torsion: beware of torsions simulating hydroceles. PMID- 8411334 TI - Varix of the left gonadal vein. PMID- 8411335 TI - Color Doppler sonography to evaluate transjugular intrahepatic portacaval stent shunt. PMID- 8411336 TI - Uterine lipoleiomyoma: a rare tumor. PMID- 8411337 TI - In vitro expression of the human cytomegalovirus DNA polymerase gene: effects of sequence alterations on enzyme activity. AB - Genomic DNA of the Towne strain human cytomegalovirus polymerase (pol) gene (4.4 kb RsrII-NcoI segment of the EcoRI J fragment) was cloned into plasmids containing either the T3 or the T7 promoter for in vitro transcription translation studies. The translation efficiency of unmodified pol cRNA was poor in this system and could not be improved by capping. However, the efficiency could be enhanced by replacing the leader sequence with a 40-bp AT-rich sequence derived from an alfalfa mosaic virus, R4. pol cRNA directed the synthesis of a 140-kDa polypeptide in a rabbit reticulocyte translation system. The in vitro translated wild-type enzyme possessed significant polymerization activity which could be stimulated by salt as could that of the authentic enzyme purified from virus-infected cells. To study the critical domains of this enzyme, nine mutations were introduced into the pol gene around the conserved domains of eukaryotic polymerase by oligonucleotide-directed mutagenesis. Two constructs with mutations at amino acid residues 323 to 325 (M32QS) and 725 to 726 (M72II) remained active, with partial loss of enzyme activity, while the enzyme activities of other mutants with alterations at four domains located around amino acid residues 729 to 730 (M73HN), 804 to 807 (M80 and DE80), 910 to 913 (M91 and DE91), and 962 to 964 (M96 and DE96) were abolished. DNA template and triphosphate binding assays indicated that the mutation at 804 to 807 (conserved region III) lost the ability to bind DNA template, and four mutants, M73HN (within conserved region II), M80 (in region III), M91 (in region I), and M96 (around region V [962 to 964; amino acid sequence KKR]), failed to bind deoxyribonucleoside triphosphate. These data suggest that conserved region III is essential for DNA template binding, while residues between conserved region II and V (725 to 964) are involved in triphosphate binding. PMID- 8411338 TI - Alternative splicing of human immunodeficiency virus type 1 mRNA modulates viral protein expression, replication, and infectivity. AB - Multiple RNA splicing sites exist within human immunodeficiency virus type 1 (HIV 1) genomic RNA, and these sites enable the synthesis of many mRNAs for each of several viral proteins. We evaluated the biological significance of the alternatively spliced mRNA species during productive HIV-1 infections of peripheral blood lymphocytes and human T-cell lines to determine the potential role of alternative RNA splicing in the regulation of HIV-1 replication and infection. First, we used a semiquantitative polymerase chain reaction of cDNAs that were radiolabeled for gel analysis to determine the relative abundance of the diverse array of alternatively spliced HIV-1 mRNAs. The predominant rev, tat, vpr, and env RNAs contained a minimum of noncoding sequence, but the predominant nef mRNAs were incompletely spliced and invariably included noncoding exons. Second, the effect of altered RNA processing was measured following mutagenesis of the major 5' splice donor and several cryptic, constitutive, and competing 3' splice acceptor motifs of HIV-1NL4-3. Mutations that ablated constitutive splice sites led to the activation of new cryptic sites; some of these preserved biological function. Mutations that ablated competing splice acceptor sites caused marked alterations in the pool of virus-derived mRNAs and, in some instances, in virus infectivity and/or the profile of virus proteins. The redundant RNA splicing signals in the HIV-1 genome and alternatively spliced mRNAs provides a mechanism for regulating the relative proportions of HIV-1 proteins and, in some cases, viral infectivity. PMID- 8411339 TI - Marked, transient inhibition of expression of the Epstein-Barr virus latent membrane protein gene in Burkitt's lymphoma cell lines by electroporation. AB - The Raji, EB1, and EB2 cell lines are derived from Epstein-Barr virus (EBV) positive Burkitt's lymphomas. EBV gene products associated with viral latency, including latent membrane protein (LMP) and Epstein-Barr nuclear antigen 2 (EBNA 2), are expressed in these cell lines. We have found that transfection of all three of these cell lines by electroporation in both the presence and the absence of foreign DNA resulted in a marked decrease in expression of the endogenous EBV gene encoding LMP. An analysis of this response in Raji cells revealed that the level of RNA of this gene was decreased transiently and returned to normal levels by 7 days after transfection. The level of LMP protein was also reduced after transfection. No difference in survival was detected in electroporated versus unperturbed Raji cells. The level of mRNA encoding a modulator of the LMP gene, EBNA-2, was unchanged by electroporation. However, the level of another EBV transcript, BHLF-I, was reduced. The effect of electroporation could not be attributed to flux of Ca2+, Na+, K+, or Cl- ions across the plasma membrane. Expression of LMP in several lymphoblastoid cell lines was unaffected by electroporation. PMID- 8411340 TI - Mutations in the N-terminal region of human immunodeficiency virus type 1 matrix protein block intracellular transport of the Gag precursor. AB - The matrix domain of human immunodeficiency virus type 1 Gag polyprotein was studied for its role in virus assembly. Deletion and substitution mutations caused a dramatic reduction in virus production. Mutant Gag polyproteins were myristoylated and had a high affinity for membrane association. Immunofluorescence staining revealed a large accumulation of mutant Gag precursors in the cytoplasm, while wild-type Gag proteins were primarily associated with the cell surface membrane. These results suggest a defect in intracellular transport of the mutant Gag precursors. Thus, in addition to myristoylation, the N-terminal region of the matrix domain is involved in determining Gag protein transport to the plasma membrane. Wild-type Gag polyproteins interacted with and efficiently packaged mutant Gag into virions. This finding is consistent with the hypothesis that intermolecular interaction of Gag polyproteins might occur in the cytoplasm prior to being transported to the assembly site on the plasma membrane. PMID- 8411341 TI - Characterization of bovine immunodeficiency virus rev cDNAs and identification and subcellular localization of the Rev protein. AB - One of the six putative accessory genes of bovine immunodeficiency virus (BIV) is similar to those identified as rev in the human immunodeficiency virus and visna virus genomes. To further analyze the BIV rev gene locus, protein, and function, rev cDNAs were cloned and characterized. BIV rev mRNA is derived from the full length transcript by multiple splicing events and consists of three exons, including the untranslated leader sequence and two coding exons. BIV rev cDNA was expressed in bacteria and in a mammalian in vitro translation expression system. A 23-kDa Rev protein (p23rev) was immunologically detected in lysates from both systems by using an antiserum made to a synthetic Rev peptide. Recombinant p23rev made in bacteria was purified and used to make a polyvalent antiserum. Antisera to Rev peptide and recombinant p23rev immunoprecipitated p23rev from BIV-infected mammalian cells but not from virions. A mammalian expression vector using the BIV rev cDNA was constructed; p23rev was immunoprecipitated with anti-Rev serum from 32P-labeled lysates of monkey cells transfected with this plasmid, demonstrating that BIV Rev is phosphorylated. Immunofluorescence and immunoelectron microscopy with anti-BIV Rev antisera localized Rev in the nucleus and, particularly, in the nucleoli of BIV-infected cells. In functional studies, the expression of BIV Rev was shown to positively regulate the appearance both of Gag protein, which is translated from the unspliced primary viral transcript, and of singly spliced env mRNA but not that of the multiply spliced tat mRNA. These results demonstrate that BIV Rev activity correlates with the known function of lentivirus Rev proteins. PMID- 8411342 TI - Identification of domains in brome mosaic virus RNA-1 and coat protein necessary for specific interaction and encapsidation. AB - Even though many single-stranded RNAs are present in the cytoplasm of infected cells, encapsidation by brome mosaic virus (BMV) coat protein is specific for BMV RNA. Although the highly conserved 3' region of each of the three BMV genomic RNAs is an attractive candidate for the site of recognition by the coat protein, band shift and UV cross-linking assays in the presence of specific and nonspecific competitors revealed only nonspecific interactions. However, BMV RNA 1 formed a retarded complex (complex I) with the coat protein in the absence of competitors, and two domains of RNA-1 that specifically bound coat protein in a small complex (complex II), presumably early in the encapsidation process, were identified. Strong nonspecific, cooperative binding was observed in the presence of high concentrations of coat protein, suggesting that this provides the mechanism leading to rapid encapsidation seen in vivo. In contrast, no binding to a coat protein mutant lacking the N-terminal 25 amino acids that has been shown to be incapable of encapsidation in vivo (R. Sacher and P. Ahlquist, J. Virol. 63:4545-4552, 1989) was detected in vitro. The use of deletion mutants of RNA-1 revealed the presence of domains within the coding region of protein 1a that formed complexes with purified coat protein. One deletion mutant (B1SX) lacking these domains was only slightly more effective in dissociating RNA-1-coat protein complexes than were nonspecific competitors, further suggesting that regions other than the 3' end can participate in the selective encapsidation of BMV RNAs. PMID- 8411343 TI - The genetic drift of human papillomavirus type 16 is a means of reconstructing prehistoric viral spread and the movement of ancient human populations. AB - We have investigated the diversity of a hypervariable segment of the human papillomavirus type 16 (HPV-16) genome among 301 virus isolates that were collected from 25 different ethnic groups and geographic locations. Altogether, we distinguished 48 different variants that had diversified from one another along five phylogenetic branches. Variants from two of these branches were nearly completely confined to Africa. Variants from a third branch were the only variants identified in Europeans but occurred at lower frequency in all other ethnic groups. A fourth branch was specific for Japanese and Chinese isolates. A small fraction of all isolates from Asia and from indigenous as well as immigrant populations in the Americas formed a fifth branch. Important patterns of HPV-16 phylogeny suggested coevolution of the virus with people of the three major human races, namely, Africans, Caucasians, and East Asians. But several minor patterns are indicative of smaller bottlenecks of viral evolution and spread, which may correlate with the migration of ethnic groups in prehistoric times. The colonization of the Americas by Europeans and Africans is reflected in the composition of their HPV-16 variants. We discuss arguments that today's HPV-16 genomes represent a degree of diversity that evolved over a large time span, probably exceeding 200,000 years, from a precursor genome that may have originated in Africa. The identification of molecular variants is a powerful epidemiological and phylogenetic tool for revealing the ancient spread of papillomaviruses, whose trace through the world has not yet been completely lost. PMID- 8411344 TI - Evolution of human papillomavirus type 18: an ancient phylogenetic root in Africa and intratype diversity reflect coevolution with human ethnic groups. AB - Papillomaviruses are an ideal model system for the study of DNA virus evolution. On several levels, phylogenetic trees of papillomaviruses reflect the relationship of their hosts. Papillomaviruses isolated from remotely related vertebrates form major branches. One branch of human papillomaviruses (HPVs) includes an ape and two monkey papillomaviruses, possibly because the diversification of the viruses predated the separation of the infected-primate taxa. This hypothesis predicts that the root of the evolution of some if not all HPV types should point to Africa, since humans evolved from nonhuman primates in this continent. We tested this hypothesis and compared the genomic sequences of HPV type 18 (HPV-18) isolates from four continents. Diversity within HPV-18 correlates with patterns of the evolution and spread of Homo sapiens: HPV-18 variants, just like HPV-16 variants, are specific for the major human races, with maximal diversity in Africa. Outgroup rooting of the HPV-18 tree against HPV-45, which is closely related to HPV-18, identifies African HPV-18 variants at the root of the tree. The identification of an African HPV-45 isolate further reduces the evolutionary distance between HPV-18 and HPV-45. HPV-18 variants from Amazonian Indians are the closest relatives to those from Japanese and Chinese patients and suggest that a single point mutation in the phylogenetically evaluated genomic segment represents at least 12,000 years of evolution. We estimate that diversity within HPV-18 and probably within other HPV types evolved over a period of more than 200,000 years and that diversity between HPV types evolved over several million years. PMID- 8411345 TI - Two amino acid substitutions in the tomato mosaic virus 30-kilodalton movement protein confer the ability to overcome the Tm-2(2) resistance gene in the tomato. AB - The Tm-2(2) resistance gene is used in most commercial tomato cultivars for protection against infection with tobacco mosaic virus and its close relative tomato mosaic virus (ToMV). To study the mechanism of this resistance gene, cDNA clones encompassing the complete genome of a ToMV strain (ToMV-2(2)) that was able to break the Tm-2(2) resistance were generated. Chimeric full-length viral cDNA clones were constructed under the control of the cauliflower mosaic virus 35S RNA promoter, combining parts of the wild-type virus and ToMV-2(2). Using these clones in cDNA infection experiments, we showed that the 30-kDa movement protein of ToMV-2(2) is responsible for overcoming the Tm-2(2) resistance gene in the tomato. DNA sequence analysis revealed four amino acid exchanges between the 30-kDa proteins from wild-type ToMV and ToMV-2(2), Lys-130 to Glu, Gly-184 to Glu, Ser-238 to Arg, and Lys-244 to Glu. To clarify the involvement of the altered amino acid residues in the resistance-breaking properties of the ToMV 2(2) movement protein, different combinations of these amino acid exchanges were introduced in the genome of wild-type ToMV. Only one mutant strain which contained two amino acid substitutions, Arg-238 and Glu-244, was able to multiply in Tm-2(2) tomato plants. Both amino acid exchanges are found within the carboxy terminal region of the movement protein, which displays a high variability among different tobamoviruses and has been shown to be dispensable for virus transport in tobacco plants. These observations suggest that the resistance conferred by the Tm-2(2) gene against ToMV depends on specific recognition events in this host pathogen interaction rather than interfering with fundamental functions of the 30 kDa protein. PMID- 8411346 TI - Sindbis virus expression vectors: packaging of RNA replicons by using defective helper RNAs. AB - Since the recovery of infectious RNA transcripts from full-length cDNA clones, alphavirus genome RNAs have been engineered to allow expression of heterologous RNAs and proteins. The highest levels of expression of heterologous products are achieved when the viral structural genes are replaced by the heterologous coding sequences. Such recombinant RNAs are self-replicating (replicons) and can be introduced into cells as naked RNA, but they require trans complementation to be packaged and released from cells as infectious virion particles. In this report, we describe a series of defective Sindbis virus helper RNAs which can be used for packaging Sindbis virus RNA replicons. The defective helper RNAs contain the cis acting sequences required for replication as well as the subgenomic RNA promoter which drives expression of the structural protein genes. In cells cotransfected with both the replicon and defective helper RNAs, viral nonstructural proteins translated from the replicon RNA allow replication and transcription of the defective helper RNA to produce the virion structural proteins. A series of defective helper RNAs were compared for the ability to package the replicon RNA as well as for the ability to be replicated and packaged. One defective helper RNA not only packaged the replicon but also was itself encapsidated and would be useful under conditions in which extensive amplification is advantageous. Other defective helper RNAs were able to package the replicon efficiently but were packaged very poorly themselves. These helpers should be useful for applications in which expression of the viral structural proteins or virus spread is not desired. PMID- 8411347 TI - Productive infection of a cervical epithelial cell line with human immunodeficiency virus: implications for sexual transmission. AB - The human cervix-derived epithelial cell line (ME180) used in this study displays a characteristics epithelial morphology, including numerous desmosomes, tonofilaments, and epidermal filaments. When T-cell lines infected with human immunodeficiency virus (HIV) are added to epithelial cultures, they rapidly adhere to the epithelial monolayer. Within a few minutes, the T cells shed numerous virions into narrow spaces formed between the epithelial cell and the adherent T cells. Virions subsequently enter the ME180 cells via large vesicles. A few days after infection, cytopathic effects and syncytium formation were observed. Infected clones of ME180 cells have remained infected for 8 months. p24 enzyme-linked immunosorbent assay and infectivity assays show that one subclone of the cell line produces virus titers equivalent to those of high-secreting HIV infected T-cell lines. Electron microscopy reveals numerous virions budding from both the basal and apical surfaces of the epithelium. These observations suggest that cervical epithelium has the potential to serve as a site of HIV infection. PMID- 8411348 TI - Molecular and biochemical bases for activation of the transforming potential of the proto-oncogene c-ros. AB - The transforming gene of avian sarcoma virus UR2, v-ros, encodes a receptor-like protein tyrosine kinase and differs from its proto-oncogene, c-ros, in its 5' truncation and fusion to viral gag, a three-amino-acid (aa) insertion in the transmembrane (TM) domain, and changes in the carboxyl region. To explore the basis for activation of the c-ros transforming potential, various c-ros retroviral vectors containing those changes were constructed and studied for their biological and biochemical properties. Ufcros codes for the full-length c ros protein of 2,311 aa, Uppcros has 1,661-aa internal deletion in the extracellular domain, CCros contains the 3' c-ros cDNA fused 150 aa upstream of the TM domain to the UR2 gag, CVros is the same as CCros except that the 3' region is replaced by that of v-ros, and VCros is the same as CCros except that the 5' region is replaced by that of v-ros. The Ufcros, Uppcros, CCros, and CVros are inactive in transforming chicken embryo fibroblasts, whereas VCros is as potent as UR2 in cell-transforming and tumorigenic activities. Upon passages of CCros and CVros viruses, the additional extracellular sequence in comparison with that of v-ros was delected; concurrently, both viruses (named CC5d and CV5d, respectively) attained moderate transforming activity, albeit significantly lower than that of UR2 or VCros. The native c-ros protein has a very low protein tyrosine kinase activity, whereas the ppcros protein is constitutively activated in kinase activity. The inability of CCros and CVros to transform chicken embryo fibroblasts is consistent with the inefficient membrane association, instability, and low kinase activity of their encoded proteins. The CC5d and CV5d proteins are indistinguishable in kinase activity, membrane association, and stability from the v-ros protein. The reduced transforming potency of CC5d and CV5d proteins can be attributed only to their differential substrate interaction, notably the failure to phosphorylate a 88-kDa protein. We conclude that the 5' rather than the 3' modification of c-ros is essential for its oncogenic activation; the sequence upstream of the TM domain has a negative effect on the transforming activity of CCros and CVros and needs to be deleted to activate their biological activity. PMID- 8411349 TI - Relative affinity of the human parainfluenza virus type 3 hemagglutinin neuraminidase for sialic acid correlates with virus-induced fusion activity. AB - The ability of enveloped viruses to cause disease depends on their ability to enter the host cell via membrane fusion events. An understanding of these early events in infection, crucial for the design of methods of blocking infection, is needed for viruses that mediate membrane fusion at neutral pH, such as paramyxoviruses and human immunodeficiency virus. Sialic acid is the receptor for the human parainfluenza virus type 3 (HPF3) hemagglutinin-neuraminidase (HN) glycoprotein, the molecule responsible for binding of the virus to cell surfaces. In order for the fusion protein (F) of HPF3 to promote membrane fusion, the HN must interact with its receptor. In the present report, two variants of HPF3 with increased fusion-promoting phenotypes were selected and used to study the function of the HN glycoprotein in membrane fusion. Increased fusogenicity correlated with single amino acid changes in the HN protein that resulted in increased binding of the variant viruses to the sialic acid receptor. These results suggest that the avidity of binding of the HN protein to its receptor regulates the level of F protein-mediated fusion and begin to define one role of the receptor-binding protein of a paramyxovirus in the membrane fusion process. PMID- 8411350 TI - Cell fusion mediated by interaction of a hybrid CD4.CD8 molecule with the human immunodeficiency virus type 1 envelope glycoprotein does occur after a long lag time. AB - Several domains of CD4 have been suggested to play a critical role in events that follow its binding to the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (gp120-gp41). It has been reported previously that cells expressing a chimeric molecule consisting of the first 177 residues of human CD4 attached to residues from the hinge, transmembrane, and cytoplasmic domains of human CD8 did not form syncytia with HIV-1-infected cells (L. Poulin, L.A. Evans, S. Tang, A. Barboza, H. Legg, D.R. Littman, and J.A. Levy, J. Virol. 65: 4893-4901, 1991). In contrast, we found that the hybrid CD4.CD8 molecule expressed in human cells did render them susceptible to fusion with cells expressing HIV-1IIIB or HIV-1RF envelope glycoproteins encoded by vaccinia virus recombinants, but only after long lag times. The lag time of membrane fusion mediated by the hybrid CD4.CD8 molecule was fivefold longer than that for the wild-type CD4 molecule. However, the rate of binding to and the affinity of soluble gp120 for membrane-associated CD4.CD8 were the same as for CD4. Both molecules were laterally mobile, as determined by patching experiments. Coexpression of the CD4.CD8 chimera with wild type CD4 did not lead to interference in fusion but had an additive effect. Therefore, the proximal membrane domains of CD4 play an important role in determining the kinetics of postbinding events leading to membrane fusion. We hypothesize that the long lag time is due to the inability of the CD4.CD8-gp120 gp41 complex to undergo the rapid conformational changes which occur during the fusion mediated by wild-type CD4. PMID- 8411351 TI - Transcriptional control of human papillomavirus (HPV) oncogene expression: composition of the HPV type 18 upstream regulatory region. AB - The malignant transformation potential of high-risk human papillomaviruses (HPVs) is closely linked to the expression of the viral E6 and E7 genes. To elucidate the molecular mechanisms resulting in HPV oncogene expression, a systematic analysis of the cis-regulatory elements within the HPV type 18 (HPV18) upstream regulatory region (URR) which regulate the activity of the E6/E7 promoter was performed. As the functional behavior of a given cis-regulatory element can be strongly influenced by the overall composition of a transcriptional control region, individual elements were inactivated by site-directed mutagenesis in the physiological context of the complete HPV18 URR. Subsequently, the effects of these mutations on the activity of the E6/E7 promoter were assessed by transient transfection assays. We found that the transcriptional stimulation of the E6/E7 promoter largely depends on the integrity of cis-regulatory elements bound by AP1, SP1, and in certain epithelial cells, KRF-1. In contrast to previous reports by implying a key role for NF1 and Oct-1 recognition motifs in the stimulation of papillomavirus oncogene expression, the inactivation of these elements in the context of the HPV18 URR did not strongly affect the transcriptional activity of the E6/E7 promoter. Mutation of a promoter-proximal glucocorticoid response element completely abolished dexamethasone inducibility of the HPV18 E6/E7 promoter and resulted in an increase of its basal activity. Functional dissection of the HPV18 constitutive enhancer region indicates that its transcriptional activity is largely generated by functional synergism between a centrally located AP1 module and thus far undetected cis-active elements present in the 5' flank of the enhancer. Furthermore, comparative analyses using homologous and heterologous promoters show that the transcriptional activity of HPV18 enhancer elements is influenced by the nature of the test promoter in a cell-type-specific manner. PMID- 8411352 TI - Functional chimeras of the Rous sarcoma virus and human immunodeficiency virus gag proteins. AB - The Gag protein encoded by Rous sarcoma virus (RSV) is the only viral product required for the process of budding whereby virus particles are formed at the plasma membrane. Deletion analysis of this Gag molecule has revealed several regions (assembly domains) that are important for budding. One of these domains is located at the amino terminus and is needed for membrane binding. Another is located within the carboxy-terminal third of the protein. Though there is little sequence homology among the Gag proteins of unrelated retroviruses, it seemed possible that their assembly domains might be functionally conserved, and to explore this idea, numerous Gag chimeras were made. The results indicate that the first 10 amino acids of the human immunodeficiency virus (HIV) Gag protein can suppress the block to budding caused by deletions in the RSV MA sequence, much as described previously for the first 10 residues from the Src oncoprotein (J.W. Wills, R.C. Craven, R. A. Weldon, Jr., T. D. Nelle, and C.R. Erdie, J. Virol. 65:3804-3812, 1991). In addition, the carboxy-terminal half of the HIV Gag protein was fused to a truncated RSV Gag molecule, mutant Bg-Bs, which is unable to direct core assembly. This chimera was able to produce particles at a rate identical to that of RSV and of a density similar to that of authentic virions. Deletion analysis of the carboxy-terminal chimera revealed two small regions within the HIV NC protein that were sufficient for endowing mutant Bg-Bs with these properties. Chimeras lacking both regions produced particles of a low density, suggesting that these sequences may be involved in the tight packing of Gag molecules during assembly. In a related set of experiments, replacement of the RSV protease with that of HIV resulted in premature processing within the RSV sequence and a block to budding. Particle assembly was restored when the HIV PR activity was inactivated by mutagenesis. Collectively, the data presented here illustrate the functional similarities of Gag proteins from unrelated retroviruses. PMID- 8411353 TI - Incorporation of human immunodeficiency virus type 1 Gag proteins into murine leukemia virus virions. AB - The retroviral Gag polyprotein is necessary and sufficient for assembly and budding of viral particles. However, the exact inter- and intramolecular interactions of the Gag polyproteins during this process are not known. To locate functional domains within Gag, we generated chimeric proviruses between human immunodeficiency virus type 1 (HIV-1) and murine leukemia virus (MuLV). In these chimeric proviruses, the matrix or capsid proteins of MuLV were precisely replaced with the matrix or capsid proteins of HIV-1. Although the chimeric proviruses were unable to efficiently assemble into mature viral particles by themselves, coexpression of wild-type MuLV Gag rescued the HIV proteins into virions. The specificity of the rescue of HIV proteins into MuLV virions shows that specific interactions involving homologous matrix or capsid regions of Gag are necessary for retroviral particle formation. PMID- 8411354 TI - Papain-like protease p29 as a symptom determinant encoded by a hypovirulence associated virus of the chestnut blight fungus. AB - Viral double-stranded RNAs (dsRNAs) responsible for virulence attenuation (hypovirulence) of the chestnut blight fungus, Cryphonectria parasitica, profoundly influence a range of host functions in addition to virulence. The 5' proximal open reading frame, A, of the prototypical hypovirulence-associated viral dsRNA, L-dsRNA, present in hypovirulent strain EP713, was recently shown by DNA-mediated transformation analysis to suppress fungal sporulation, pigmentation, and accumulation of the enzyme laccase (G. H. Choi and D. L. Nuss, EMBO J. 11:473-477, 1992). We mapped this suppressive activity to the autocatalytic papain-like protease, p29, present within the amino-terminal portion of open reading frame A-encoded polyprotein p69. Mutational analysis revealed that the ability of p29 to alter fungal phenotype is dependent upon release from the polyprotein precursor but is independent of intrinsic proteolytic activity. Deletion of the p29-coding domain within the context of an infectious L-dsRNA cDNA clone resulted in a replication-competent viral dsRNA that exhibited intermediate suppressive activity while retaining the ability to confer hypovirulence. Thus, p29 is necessary but not sufficient for the level of virus-mediated suppression of fungal pigmentation, sporulation, and laccase accumulation observed for wild-type hypovirulent strain EP713 and is nonessential for viral RNA replication and virulence attenuation. These results also illustrate the feasibility of engineering infectious viral cDNA for construction of hypovirulent fungal strains with specific phenotypic traits. PMID- 8411355 TI - Analysis of simian immunodeficiency virus sequence variation in tissues of rhesus macaques with simian AIDS. AB - One rhesus macaque displayed severe encephalomyelitis and another displayed severe enterocolitis following infection with molecularly cloned simian immunodeficiency virus (SIV) strain SIVmac239. Little or no free anti-SIV antibody developed in these two macaques, and they died relatively quickly (4 to 6 months) after infection. Manifestation of the tissue-specific disease in these macaques was associated with the emergence of variants with high replicative capacity for macrophages and primary infection of tissue macrophages. The nature of sequence variation in the central region (vif, vpr, and vpx), the env gene, and the nef long terminal repeat (LTR) region in brain, colon, and other tissues was examined to see whether specific genetic changes were associated with SIV replication in brain or gut. Sequence analysis revealed strong conservation of the intergenic central region, nef, and the LTR. However, analysis of env sequences in these two macaques and one other revealed significant, interesting patterns of sequence variation. (i) Changes in env that were found previously to contribute to the replicative ability of SIVmac for macrophages in culture were present in the tissues of these animals. (ii) The greatest variability was located in the regions between V1 and V2 and from "V3" through C3 in gp120, which are different in location from the variable regions observed previously in animals with strong antibody responses and long-term persistent infection. (iii) The predominant sequence change of D-->N at position 385 in C3 is most surprising, since this change in both SIV and human immunodeficiency virus type 1 has been associated with dramatically diminished affinity for CD4 and replication in vitro. (iv) The nature of sequence changes at some positions (146, 178, 345, 385, and "V3") suggests that viral replication in brain and gut may be facilitated by specific sequence changes in env in addition to those that impart a general ability to replicate well in macrophages. These results demonstrate that complex selective pressures, including immune responses and varying cell and tissue specificity, can influence the nature of sequence changes in env. PMID- 8411356 TI - Alternate pathways of secretion of simian immunodeficiency virus envelope glycoproteins. AB - A biotinylation assay was used to detect the envelope glycoprotein of the simian immunodeficiency virus (SIV) envelope glycoprotein expressed by a recombinant vaccinia virus on the surface of HeLa T4 cells. The relationship between the detection of the envelope glycoprotein on the cell surface and its secretion from the cell was examined. It was found that much more gp120 was released into the culture medium than could be accounted for by shedding of the biotinylated SIV envelope protein from the cell surface. Treatment with the ionophore monensin showed that this drug did not block the secretion of gp120 into the culture medium even though the expression of gp120 on the cell surface was strongly downregulated. Similar results were observed for the secretion of gp120 in HUT78 cells infected with SIVmac251 virus. Brefeldin A, on the other hand, inhibited both the detection of gp120 on the cell surface and its secretion into the culture medium. On the basis of these results, we propose that gp120 can be secreted into the culture medium via at least two pathways. One pathway involves the dissociation of gp120 from membrane-associated gp41-gp120 complexes on the cell surface. However, the major pathway involves the secretion of gp120 without its transitory appearance on the cell surface as part of a gp41-gp120 complex. PMID- 8411357 TI - Human immunodeficiency virus type 1 viral protein R localization in infected cells and virions. AB - The subcellular localization of human immunodeficiency virus type 1 (HIV-1) viral protein R (Vpr) was examined by subcellular fractionation. In HIV-1-infected peripheral blood mononuclear cells, Vpr was found in the nuclear and membrane fractions as well as the conditioned medium. Expression of Vpr without other HIV 1 proteins, in two different eukaryotic expression systems, demonstrated a predominant localization of Vpr in the nuclear matrix and chromatin extract fractions. Deletion of the carboxyl-terminal 19-amino-acid arginine-rich sequence impaired Vpr nuclear localization. Indirect immunofluorescence confirmed the nuclear localization of Vpr and also indicated a perinuclear location. Expression of Vpr alone did not result in export of the protein from the cell, but when coexpressed with the Gag protein, Vpr was exported and found in virus-like particles. A truncated Gag protein, missing the p6 sequence and a portion of the p9 sequence, was incapable of exporting Vpr from the cell. Regulation of Vpr localization may be important in the influence of this protein on virus replication. PMID- 8411358 TI - Simian virus 40 large T antigen associates with cyclin A and p33cdk2. AB - In this paper we provide evidence that a fraction of large T antigen of simian virus 40 (SV40) interacts with cyclin A and p33cdk2 in both virus-infected and stably transformed cells. Immunoprecipitates of SV40 large T antigen from SV40 infected or SV40 large-T-antigen-transformed cells contain cyclin A, p33cdk2, and histone H1 kinase activity. Conversely, immunoprecipitates of cyclin A from these cells contain SV40 large T antigen. In this respect, SV40 large T antigen has properties similar to those of the E1A oncogene of adenoviruses and the E7 oncogene of human papillomaviruses. PMID- 8411359 TI - Detection of DNA polymerase activities associated with purified duck hepatitis B virus core particles by using an activity gel assay. AB - Replication of hepadnaviruses involves reverse transcription of an intermediate RNA molecule. It is generally accepted that this replication scheme is carried out by a virally encoded, multifunctional polymerase which has DNA-dependent DNA polymerase, reverse transcriptase, and RNase H activities. Biochemical studies of the polymerase protein(s) have been limited by the inability to purify useful quantities of functional enzyme from virus particles and, until recently, to express enzymatically active polymerase proteins in heterologous systems. An activity gel assay which detects in situ catalytic activities of DNA polymerases after electrophoresis in partially denaturing polyacrylamide gels was used by M.R. Bavand and O. Laub (J. Virol. 62:626-628, 1988) to show the presence of DNA- and RNA-dependent DNA polymerase activities associated with hepatitis B virus particles produced in vitro. This assay has provided the only means by which hepadnavirus polymerase proteins have been detected in association with enzymatic activities. Since conventional methods have not allowed purification of useful quantities of enzymatically active polymerase protein(s), we have devised a protocol for purifying large quantities of duck hepatitis B virus (DHBV) core particles to near homogeneity. These immature virus particles contain DNA- and RNA-dependent DNA polymerase activities, as shown in the endogenous DNA polymerase assay. We have used the activity gel assay to detect multiple DNA- and RNA-dependent DNA polymerase proteins associated with these purified DHBV core particles. These enzymatically active proteins appear larger than, approximately the same size as, and smaller than an unmodified DHBV polymerase protein predicted from the polymerase open reading frame. This is the first report of the detection of active hepadnavirus core-associated DNA polymerase proteins derived from a natural host. PMID- 8411360 TI - Genetic determinants of dengue type 4 virus neurovirulence for mice. AB - Mouse-adapted dengue type 4 virus (DEN4) strain H241 is highly neurovirulent for mice, whereas its non-mouse-adapted parent is rarely neurovirulent. The genetic basis for the neurovirulence of the mouse-adapted mutant was studied by comparing intratypic chimeric viruses that contained the three structural protein genes from the parental virus or the neurovirulent mutant in the background sequence of nonneurovirulent DEN4 strain 814669. The chimera that contained the three structural protein genes from mouse neurovirulent DEN4 strain H241 proved to be highly neurovirulent in mice, whereas the chimera that contained the corresponding genes from its non-mouse-adapted parent was not neurovirulent. This finding indicates that most of the genetic loci for the neurovirulence of the DEN4 mutant lie within the structural protein genes. A comparison of the amino acid sequences of the parent and its mouse neurovirulent mutant proteins revealed that there were only five amino acid differences in the structural protein region, and three of these were located in the envelope (E) glycoprotein. Analysis of chimeras which contained one or two of the variant amino acids of the mutant E sequence substituting for the corresponding sequence of the parental virus identified two of these amino acid changes as important determinants of mouse neurovirulence. First, the single substitution of Ile for Thr-155 which ablated one of the two conserved glycosylation sites in parental E yielded a virus that was almost as neurovirulent as the mouse-adapted mutant. Thus, the loss of an E glycosylation site appears to play a role in DEN4 neurovirulence. Second, the substitution of Leu for Phe-401 also yielded a neurovirulent virus, but it was less neurovirulent than the glycosylation mutant. These findings indicate that at least two of the genetic loci responsible for DEN4 mouse neurovirulence map within the structural protein genes. PMID- 8411361 TI - Protein interactions with DNA elements in variant equine infectious anemia virus enhancers and their impact on transcriptional activity. AB - The long terminal repeats (LTRs) from various cloned equine infectious anemia virus (EIAV) proviruses differ significantly, but all contain cis-acting DNA elements identical to MDBP-, PEA2-, AP-1-, and PU.1 (ets)-binding sites. A prototype EIAV LTR would contain one of each of these conserved elements. The LTR variations originate from the insertion of novel sequences between the PEA2 and AP-1 elements in the transcriptional enhancer unit. Viewed in this way, the LTR from provirus clone lambda 12 has an 11-bp insertion containing a PEA2 site and the LTR of the lambda 6 provirus has a 31-bp insertion/duplication containing PEA2, AP-1, and PU.1 sites. Two other LTRs were cloned by amplification of cDNAs from the persistently infected cell line, EIAV-FEA. A third LTR was generated by site-directed mutagenesis of one of the LTRs from EIAV-FEA cells. The latter three had a single base change in the element next to the TATA box that abolished PU.1 binding; however, the variable regions of these LTRs were shown by gel mobility shift assays to contain one or two PU.1 sites. One variable region was shown to have an octamer site overlapping its tandem PU.1 elements. Basal, PMA activated, and Tat trans-activated transcriptional activities of the LTRs were compared in several different cell lines by transient transfection. The various promoters displayed different relative levels of activity depending on the cell line used and the condition of activation. This natural set of variant promoters may help define how changes in the components of the transcription complex influence transactivation by Tat. The diverse LTRs could endow their respective proviruses with a unique pattern of expression and activation in vivo. PMID- 8411362 TI - Phosphatidylcholine hydrolysis activates NF-kappa B and increases human immunodeficiency virus replication in human monocytes and T lymphocytes. AB - We have tested whether breakdown of phosphatidylcholine (PC) initiated by exogenous addition of a PC-specific phospholipase C (PC-PLC) from Bacillus cereus or by endogenous overexpression of PC-PLC induces functional activation of NF kappa B and increases human immunodeficiency virus (HIV) enhancer activity. PC PLC-activated hydrolysis of PC was found to induce bona fide p50/p65 NF-kappa B binding activity in three different cell lines of human or murine origin. No significant changes in the turnover of other cellular phospholipids were detected in PC-PLC-treated cells. Induction of NF-kappa B by PC-PLC did not depend on de novo synthesis of proteins or autocrine secretion of either tumor necrosis factor or interleukin 1. In human monocytic and lymphoblastoid T-cell lines, induction of NF-kappa B by PC-PLC resulted in clear induction of luciferase expression vectors placed under the control of synthetic kappa B enhancers or wild type, but not kappa B-mutated, HIV long terminal repeat constructs. HIV replication was increased by PC-PLC in chronically infected monocytes and T lymphocytes. NF-kappa B activation promoted by addition of exogenous PC-PLC correlated with an intense production of diacylglycerol. However, addition of a phosphatidylinositol specific PLC from B. cereus also induced diacylglycerol but did not activate kappa B enhancer-directed vectors. PC-PLC-induced NF-kappa B activation could not be blocked by a specific inhibitor of phorbol ester-inducible protein kinases C. These results indicate that a cellular transduction pathway, dependent on specific PC breakdown, is functional in T lymphocytes and monocytes and may be used by various transmembrane receptors to activate HIV transcription through NF kappa B-dependent induction of the HIV enhancer. PMID- 8411363 TI - Comparative analysis of the structure and function of adenovirus virus-associated RNAs. AB - The protein kinase DAI is an important component of the interferon-induced cellular defense mechanism. In cells infected by adenovirus type 2 (Ad2), activation of the kinase is prevented by the synthesis of a small, highly ordered virus-associated (VA) RNA, VA RNAI. The inhibitory function of this RNA depends on its structure, which has been partially elucidated by a combination of mutagenesis and RNase sensitivity analysis. To gain further insight into the structure and function of this regulatory RNA, we have compared the primary sequences, secondary structures, and functions of seven VA RNA species from five human and animal adenoviruses. The sequences exhibit variable degrees of homology, with a particularly close relationship between the VA RNAII species of Ad2 and Ad7 and notably divergent sequence for the avian (CELO) virus VA RNA. Apart from two pairs of mutually complementary tetranucleotides which are highly conserved, homologies are limited to transcription signals located within the RNA sequence and at its termini. Secondary structure analysis indicated that all seven RNAs conform to the model in which VA RNA possesses three main structural regions, a terminal stem, an apical stem-loop, and a central domain, although these elements vary in size and other details. The apical stem is implicated in binding to DAI, and the central domain is essential for inhibition of DAI activation. One of the pairs of conserved tetranucleotides (CCGG:C/UCGG) provides further evidence for the existence of the apical stem, but the other conserved pair (GGGU:ACCC) strongly suggests a revised structure for the central domain. In two functional assays conducted in vivo, the VA RNAI species of Ad2 and Ad7 were the most active, their corresponding VA RNAII species displayed little activity, and the single VA RNAs of Ad12 and simian adenovirus type 7 exhibited intermediate activity. Correlation of the structural and functional data suggests that the VA RNAII species adopt a structure different from those of the other VA RNA species and may play a different role in the life cycle of the virus. PMID- 8411364 TI - Analysis of the role of the bel and bet open reading frames of human foamy virus by using a new quantitative assay. AB - We have constructed a BHK-21-derived indicator cell line containing a single integrated copy of the beta-galactosidase (beta-Gal) gene under control of the human foamy virus (HFV) long terminal repeat promoter (from -533 to +20). These foamy virus-activated beta-Gal expression (FAB) cells can be used in a quantitative assay to measure the infectious titer of HFV. Our results show that the FAB assay is 50 times more sensitive than determination of the virus titer by the end-point dilution method. Using the FAB assay, we have found that HFV can productively replicate in several erythroblastoid cell lines as well as in the Jurkat T-cell line. We have also examined the roles of bel2, bet, and bel3 in viral replication by constructing proviral HFV clones in which the reading frame of Bel2, Bet, or Bel3 is disrupted by placement of translation stop codons. Analysis of these mutants reveals that while the bel3 gene is not required for viral replication in vitro, mutations in the bel2 or bet gene decrease cell-free viral transmission approximately 10-fold. PMID- 8411365 TI - Genomic concatemerization/deletion in rotaviruses: a new mechanism for generating rapid genetic change of potential epidemiological importance. AB - Three variants of group A rotavirus with large changes in their gene 5 structures have been analyzed at the molecular level. The first of these, P9 delta 5, was obtained during plaque purification undertaken as part of the biological cloning of a field isolate of virus. The gene 5 homolog in this isolate migrated just ahead of the normal segment 6 RNA, giving an estimated size of 1,300 bp. Molecular cloning and sequencing of this homolog revealed it to have a single 308 bp deletion in the center of the normal gene 5 sequence extending between nucleotides 460 and 768 of the normal gene sequence. This deletion caused a frameshift in the gene such that a stop codon was encountered 8 amino acids downstream of the deletion point, giving a predicted size for the protein product of this gene of 150 amino acids compared with the 490 amino acids of its normal size counterpart. Attempts to detect this shortened protein in virus-infected cells were not successful, indicating that it was much less stable than the full length protein and/or had suffered a large change in its antigenicity. The second two variants, brvA and brvE, were generated in an earlier study following the high-multiplicity passage of the UKtc strain of bovine rotavirus. Polyacrylamide gel electrophoresis analysis of these nondefective variants showed that brvA had a gene 5 homolog approximately equal in size to the normal RNA segment 2 (approximately 2,700 bp) and that brvE had a size of approximately 2,300 bp. Both variants showed changes in their gene 5 protein products, with brvA mimicking P9 delta 5 in failing to produce a detectable product whereas brvE produced a new virus-specific protein approximately 80 kDa in size. Full-length cDNA clones of the brvE gene 5 homolog were isolated, and analysis of their structure revealed a head-to-tail concatemerization of the normal gene 5 sequence with the first copy of the concatemer covering nucleotides 1 to 808 and the second covering nucleotides 92 to 1579, giving a total length of 2,296 bp. Sequencing across the junction region of the two copies of the gene showed that they were joined in frame to give a predicted combined open reading frame of 728 amino acids with the amino-terminal region consisting of amino acids 1 to 258 fused at the carboxy terminus to amino acids 21 to 490.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411366 TI - Rotavirus vaccine administered parenterally induces protective immunity. AB - We performed experiments to determine whether parenteral immunization with SA11 rotavirus can induce active protective immunity in a rabbit model of rotavirus infection. After one or two intramuscular injections of 1 ml of live or formalin inactivated SA11 virus, we evaluated the mucosal and serologic immune response and protection from challenge with a high dose of live, virulent rabbit (Ala) rotavirus. Inactivated SA11 virus preparations, evaluated by enzyme-linked immunosorbent assay (ELISA) with a panel of VP4- and VP7-specific neutralizing and nonneutralizing monoclonal antibodies, did not show a loss of epitopes from the inactivation procedure compared with live virus. Administration of two doses of vaccine, one at zero days postvaccination (DPV) and a booster shot at 49 DPV, followed by challenge at 71 DPV with 3.5 x 10(5) PFU of Ala virus resulted in protection from challenge. None of the two-dose virus-vaccinated rabbits shed virus after challenge, while virus shedding was detected in all control rabbits (P = 0.001, Fisher's exact two-tailed test). Differences in total serum immunoglobulin (Ig) antirotavirus ELISA titers (P < 0.05, Wilcoxon's rank sum test) were observed between groups vaccinated with virus in aluminum phosphate or Freund's adjuvant but not between groups vaccinated with live or inactivated virus in either adjuvant. All rabbits given two doses of vaccine had detectable antirotavirus intestinal antibody of the IgG, but not IgA, isotype. After challenge, fourfold or greater increases in intestinal IgG antibody responses were observed in three rabbits, whereas all controls and all but one virus vaccinated rabbit had an intestinal IgA antibody response. In contrast, vaccination of rabbits with one dose of SA11 followed by challenge at 21 DPV did not protect from challenge; no difference in the mean number of days of virus shedding between any of the vaccinated groups and controls was observed. A serologic, but not a mucosal, antibody response was observed after the one-dose vaccination regimen. Differences in serologic antibody titers were not observed between any of the one-dose virus-vaccinated groups. These data indicate that parenteral vaccination with two, but not one, doses of rotavirus in either Freund's adjuvant or aluminum phosphate can induce active protection from challenge.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411367 TI - Identification of the infected target cell type in spongiform myeloencephalopathy induced by the neurotropic Cas-Br-E murine leukemia virus. AB - The Cas-Br-E murine leukemia virus (MuLV) induces a progressive hindlimb paralysis accompanied by a spongiform myeloencephalopathy in susceptible mice. In order to better understand the pathological process leading to these neurodegenerative lesions, we have investigated the nature of the cell type(s) infected by the virus during the course of the disease in CFW/D and SWR/J mice. For this purpose, we used in situ hybridization with virus-specific probes in combination with cell-type-specific histochemical (lectin) and immunological markers as well as morphological assessment. In the early stage of infection, endothelial cells represented the main cell type expressing viral RNA in the central nervous system (CNS). With disease progression and the appearance of lesions, microglial cells became the major cell type infected, accounting for up to 65% of the total infected cell population in diseased areas. Morphologically, these cells appeared activated and were frequently found in clusters. Infection and activation of microglial cells were almost exclusively restricted to diseased regions of the CNS. Neurons in diseased regions were not discernibly infected with virus at either early or late times of disease progression. Similarly, the proportion of infected astrocytes was typically < 1%. Although some endothelial cells and oligodendrocytes were infected by the virus, their infection was not limited to diseased CNS regions. These results are consistent with a model of indirect motor neuron degeneration, subsequent to the infection of nonneuronal CNS cells and especially of microglial cells. Infected microglial cells may play a role in the disease process by releasing not only virions or viral env-gene encoded gp70 proteins but also other factors which may be directly or indirectly toxic to neurons. Parallels between microglial cell infection by MuLV and by lentiviruses, and specifically by human immunodeficiency virus, are discussed. PMID- 8411368 TI - Glycosylation of neuraminidase determines the neurovirulence of influenza A/WSN/33 virus. AB - The neuraminidase (NA) gene of influenza A/WSN/33 (WSN) virus has previously been shown to be associated with neurovirulence in mice and growth in Madin-Darby bovine kidney (MDBK) cells. Nucleotide sequence analysis has indicated that the NA of WSN virus lacks a conserved glycosylation site at position 130 (corresponding to position 146 in the N2 subtype). To investigate the role of this carbohydrate in viral pathogenicity, we used reverse genetics methods to generate a Glyc+ mutant virus, in which the glycosylation site Asn-130 was introduced into the WSN virus NA. Unlike the wild-type WSN virus, the Glyc+ mutant virus did not undergo multicycle replication in MDBK cells in the absence of trypsin, presumably because of lack of cleavage activation of infectivity. In contrast, revertant viruses derived from the Glyc+ mutant were able to replicate in MDBK cells without exogenous protease. Nucleotide sequence analysis revealed that the NAs of the revertant viruses had lost the introduced glycosylation site. In contrast to wild-type and revertant viruses, the Glyc+ mutant virus was not able to multiply in mouse brain. These results suggest that the absence of a glycosylation site at position 130 of the NA plays a key role in the neurovirulence of WSN virus in mice. PMID- 8411369 TI - Differential effects of rabies and borna disease viruses on immediate-early- and late-response gene expression in brain tissues. AB - In situ hybridization and Northern blot analysis were used to examine expression of the immediate-early-response genes (IEGs) egr-1, junB, and c-fos, and the late response gene encoding enkephalin in the brains of rats infected intranasally with Borna disease virus (BDV) or rabies virus. In both Borna disease and rabies virus infections, a dramatic and specific induction of IEGs was detected in particular regions of the hippocampus and the cortex. Increased IEG mRNA expression overlapped with the characteristic expression patterns of BDV RNA and rabies virus RNA, although relative expression levels of viral RNA and IEG mRNA differed, particularly in the hippocampal formation. Furthermore, the temporal relationship between viral RNA synthesis and activation of IEG mRNA expression in BDV infection differed markedly from that in rabies virus infection, suggesting that IEG expression is upregulated by different mechanisms. Expression of proenkephalin (pENK) mRNA was also significantly increased in BDV infection, whereas in rabies virus infection, pENK mRNA levels and also the levels of glyceraldehyde-3-phosphate dehydrogenase mRNA were reduced at terminal stages of the disease, probably reflecting a generalized suppression of cellular protein synthesis due to massive production of rabies virus mRNA. The correlation between activated IEG mRNA expression and the strong increase in viral RNA raises the possibility that IEG products induce some phenotypic changes in neurons that render them more susceptible to viral replication. PMID- 8411370 TI - Transcriptional activation by simian virus 40 large T antigen: requirements for simple promoter structures containing either TATA or initiator elements with variable upstream factor binding sites. AB - The simian virus 40 large T antigen is a promiscuous transcriptional activator of many viral and cellular promoters. We show that the promoter structure necessary for T antigen-mediated transcriptional activation is very simple. A TATA or initiator element is required, in addition to an upstream factor-binding site, which can be quite variable. We found that promoters containing an SP1-, ATF-, AP1-, or TEF-I-binding site, in conjunction with a TATA element, can all be activated in the presence of T antigen. In addition, preference for specific TATA elements was indicated. Promoters containing the HSP70 TATA element functioned better than those with the adenovirus E2 TATA element, while promoters containing the simian virus 40 (SV40) early TATA element failed to be activated. In addition, simple promoters containing the initiator element from the terminal deoxynucleotidyltransferase gene could be activated by T antigen. The SV40 late promoter, a primary target for T antigen transcriptional activation, conforms to this simple promoter structure. The region from which most late transcripts initiate contains a cluster of initiator-like elements (SV40 nucleotides [nt] 250 to 335) forming an initiator region (IR). This lies downstream of the previously described octamer-TEF element (SV40 nt 199 to 218) which contains the TEF-I binding sites shown to be necessary for T antigen-mediated transcriptional activation of the late promoter. We show that a simple late promoter made up of IR sequences and octamer-TEF element-containing sequences is transcriptionally activated by T antigen. These experiments also showed that specific sequences in the IR, SV40 nt 272 to 294, are particularly important for late promoter activation. Previous findings (M. C. Gruda, J. M. Zablotny, J. H. Xiao, I. Davidson, and J. C. Alwine, Mol. Cell. Biol. 13:961-969, 1993) suggested that T antigen could mediate transcriptional activation through interaction with the TATA-binding protein, as well as upstream bound transcription factors. Our present data are predicted by this model and suggest that at least one mechanism by which the T antigen manifests promiscuous transcriptional activation is its ability to interact with numerous transcription factors in a simple promoter context. PMID- 8411371 TI - Efficient transcriptional activation of many simple modular promoters by simian virus 40 large T antigen. AB - Simian virus 40 (SV40) large T antigen is a multifunctional protein which plays central roles during both lytic and transforming infections by SV40. It is a potent transcriptional activator and increases expression from the SV40 late promoter and from several cellular promoters. To understand better the transcriptional activation activity of large T antigen, we examined its ability to transactivate a set of simple modular promoters containing one of four upstream activation sequences coupled with one of three different TATA box sequences originally constructed and studied by Taylor and Kingston (Mol. Cell. Biol. 10:165-175, 1990). Large T antigen activated transcription from all of these simple promoters. The identity of the TATA box was a more important determinant of the final level of gene expression than was the identity of the upstream activating sequence element. We also determined the ability of a set of mutant SV40 large T antigens to activate a subset of these promoters. Several mutant SV40 large T antigens which had reduced ability to activate the complex SV40 late and Rous sarcoma virus long terminal repeat promoters showed reduced transcriptional activation activity on all of the modular promoters tested. We used a set of promoter derivatives of the human U6 small nuclear RNA promoter containing different TATA boxes and found that wild-type large T antigen could activate transcription from all of them, although to widely different levels of expression. PMID- 8411372 TI - Replication of a macrophage-tropic strain of human immunodeficiency virus type 1 (HIV-1) in a hybrid cell line, CEMx174, suggests that cellular accessory molecules are required for HIV-1 entry. AB - To investigate the mechanism underlying one aspect of the cellular tropism of human immunodeficiency virus type 1 (HIV-1), we used a macrophage-tropic isolate, 89.6, and screened its ability to infect a number of continuous cell lines. HIV-1 (89.6) was able to replicate robustly in a T-cell/B-cell hybrid line, CEMx174, while it replicated modestly or not at all in either of its parents, one of which is the CD4-positive line CEM.3. Analysis by transfection of a molecular clone, a virus uptake assay, and polymerase chain reaction all provided strong evidence that the block to HIV-1(89.6) replication in the CEM.3 line lies at the level of cellular entry. These results were complemented by preparing a CD4-expressing derivative of the B-cell parent, 721.174, and demonstrating that it is permissive for productive HIV-1(89.6) replication. Given these experimental findings, we speculate that there exist cellular accessory factors which facilitate virus entry and infection in CD4-positive cells. Furthermore, these cellular accessory factors may be quite virus strain specific, since not all macrophage-tropic strains of HIV-1 were able to replicate in the CEMx174 hybrid cell line. This experimental model provides a system for the identification of one or more of these putative cellular accessory factors. PMID- 8411373 TI - Cell type-specific proteins which interact with the 5' nontranslated region of hepatitis A virus RNA. AB - The 5' nontranslated region (5'NTR) of hepatitis A virus (HAV) RNA contains structural elements which facilitate 5' cap-independent initiation of virus translation and are likely to interact with cellular proteins functioning as translation initiation factors. To define these interactions, we characterized the binding of ribosome-associated proteins from several cell types to synthetic RNAs representing segments of the 5'NTR by using a UV cross-linking/label transfer assay. Four major proteins (p30, p39, p57, and p110) were identified. p30 and p39 were present in ribosomal salt washes prepared only from HAV permissive BS-C-1 and FRhK-4 cells, while p57 was found only in HeLa cells and rabbit reticulocyte lysates. p110 was present in all cell types. Both p30 and p39 bound to multiple sites within the 5'NTR. Efficient transfer of label to p30 occurred with minimal RNA probes representing nucleotides (nt) 96 to 155, 151 to 354, and, to a much lesser extent, 634 to 744, while label transfer to p39 occurred with probes representing nt 96 to 155 and 634 to 744. All of these probes represent regions of the 5'NTR which are rich in pyrimidines. Competitive inhibition studies indicated that both p30 and p39 bound with greater affinity to sites in the 5' half of the NTR (a probe representing nt 1 to 354) than to the more 3' site (nt 634 to 744). Binding of p39 to the probe representing nt 96 to 155 was inhibited in the presence of an equal amount of proteins derived from HeLa cells, suggesting that p39 shares binding site specificity with one or more HeLa cell proteins. A 57-kDa protein in HeLa cell protein extracts reacted with antibody to polypyrimidine tract-binding protein in immunoblots, but no immunoreactive protein was identified in a similar BS-C-1 protein fraction. These results demonstrate that ribosome-associated proteins which bind to the 5'NTR of HAV vary substantially among different mammalian cell types, possibly accounting for differences in the extent to which individual cell types support growth of the virus. Mutations in the 5'NTR which enhance the growth of HAV in certain cell types may reflect specific adaptive responses to these or other proteins. PMID- 8411374 TI - A functional GTP-binding motif is necessary for antiviral activity of Mx proteins. AB - Mx proteins are interferon-induced GTPases that inhibit the multiplication of certain negative-stranded RNA viruses. However, it has been unclear whether GTPase activity is necessary for antiviral function. Here, we have introduced mutations into the tripartite GTP-binding consensus elements of the human MxA and mouse Mx1 proteins. The invariant lysine residue of the first consensus motif, which interacts with the beta- and gamma-phosphates of bound GTP in other GTPases, was deleted or replaced by methionine or alanine. These Mx mutants and appropriate controls were then tested for antiviral activity, GTP-binding capacity, and GTPase activity. We found a direct correlation between the GTP binding capacities and GTP hydrolysis activities of the purified Mx mutants in vitro and their antiviral activities in transfected 3T3 cells, demonstrating that a functional GTP-binding motif is necessary for virus inhibition. Our results, thus, firmly establish antiviral activity as a novel function of a GTPase, emphasizing the enormous functional diversity of GTPase superfamily members. PMID- 8411375 TI - Definition of a domain of GLVR1 which is necessary for infection by gibbon ape leukemia virus and which is highly polymorphic between species. AB - Expression of human GLVR1 in mouse cells confers susceptibility to infection by gibbon ape leukemia virus (GALV), while the normally expressed mouse Glvr-1 does not. Since human and murine GLVR1 proteins differ at 64 positions in their sequences, some of the residues differing between the two proteins are critical for infection. To identify these, a series of hybrids and in vitro-constructed mutants were tested for the ability to confer susceptibility to infection. The results indicated that human GLVR1 residues 550 to 551, located in a cluster of seven of the sites that differ between the human and mouse proteins, are the only residues differing between the two which must be in the human protein form to allow infection. Sequencing of a portion of GLVR1 from the rat (which is infectible) confirmed the importance of this cluster in that it contained the only notable differences between the rat and mouse proteins. This region, which also differs substantially between the rat and the human proteins, therefore exhibits a pronounced tendency for polymorphism. PMID- 8411376 TI - Mutation of amino acids within the gibbon ape leukemia virus (GALV) receptor differentially affects feline leukemia virus subgroup B, simian sarcoma associated virus, and GALV infections. AB - The three type C retroviruses, gibbon ape leukemia virus (GALV), simian sarcoma associated virus (SSAV), and feline leukemia virus subgroup B (FeLV-B), infect human cells by interacting with the same cell surface receptor, GLVR1. Using LacZ retroviral pseudotypes and murine cells transfected with mutant GLVR1 expression vectors, we show that the same 9-amino-acid region of human GLVR1 is critical for infection by the three viruses. Rat cells were not susceptible to infection by LacZ (FeLV-B) pseudotypes because of a block at the receptor level. We found multiple amino acid differences from human GLVR1 in the 9-amino-acid critical region of rat GLVR1. Expression of a human-rat chimeric GLVR1 in murine cells demonstrated that rat GLVR1 could function as a receptor for GALV and SSAV but not for FeLV-B. Substitution of human GLVR1 amino acids in the critical region of rat GLVR1 identified three amino acids as responsible for resistance to FeLV-B infection; two of these affect SSAV infection, but none affects GALV infection. PMID- 8411377 TI - Cell-type-specific activity of the human papillomavirus type 18 upstream regulatory region in transgenic mice and its modulation by tetradecanoyl phorbol acetate and glucocorticoids. AB - The upstream regulatory region (URR) of human papillomavirus type 18 (HPV-18) harbors transcriptional promoter and enhancer elements which are thought to determine the cell-type specificity of the virus. In order to study the regulation of HPV-18 expression in vivo, we constructed transgenic mice carrying the bacterial lacZ gene under the control of the HPV-18 URR. Analysis of beta galactosidase activity by histochemical staining of tissue sections of four independent transgenic mice showed that the viral promoter was specifically active in epithelial cells within a variety of organs (e.g., tongue, ovary, uterus, testis, and small intestine). Very strong staining was observed in newborn transgenic mice in contrast to a weak activity found during fetal life. Determination of beta-galactosidase activity in crude extracts from tissues of three lines of transgenic mice proved to be a useful tool for a quantitative analysis of transgene expression. In mice from two different transgenic lines treated with dexamethasone such measurements revealed a biphasic effect of the hormone on the activity of the enzyme in the stratified epithelium of the tongue (transient increase followed by a decrease). Northern (RNA) blot analysis showed similar changes in beta-galactosidase mRNA in that tissue. Treatment with tetradecanoyl phorbol acetate (TPA) led to a twofold increase in both enzymatic activity and mRNA levels. Finally, combined treatments with dexamethasone and TPA showed that both factors interfered with each other in their respective effects on transgene expression, suggesting that a cross-talk mechanism between transcription factors could be involved in the regulation of the HPV-18 URR. PMID- 8411378 TI - Characterization of hepatitis C virus envelope glycoprotein complexes expressed by recombinant vaccinia viruses. AB - We constructed recombinant vaccinia virus vectors for expression of the structural region of hepatitis C virus (HCV). Infection of mammalian cells with a vector (vv/HCV1-906) encoding C-E1-E2-NS2 generated major protein species of 22 kDa (C), 33 to 35 kDa (E1), and 70 to 72 kDa (E2), as observed previously with other mammalian expression systems. The bulk of the E1 and E2 expressed by vv/HCV1-906 was found integrated into endoplasmic reticulum membranes as core glycosylated species, suggesting that these E1 and E2 species represent intracellular forms of the HCV envelope proteins. HCV E1 and E2 formed E1-E2 complexes which were precipitated by either anti-E1 or anti-E2 serum and which sedimented at approximately 15 S on glycerol density gradients. No evidence of intermolecular disulfide bonding between E1 and E2 was detected. E1 and E2 were copurified to approximately 90% purity by mild detergent extraction followed by chromatography on Galanthus nivalus lectin-agarose and DEAE-Fractogel. Immunization of chimpanzees with purified E1-E2 generated high titers of anti-E1 and anti-E2 antibodies. Further studies, to be reported separately, demonstrated that purified E1-E2 complexes were recognized at high frequency by HCV+ human sera (D. Y. Chien, Q.-L. Choo, R. Ralston, R. Spaete, M. Tong, M. Houghton, and G. Kuo, Lancet, in press) and generated protective immunity in chimpanzees (Q.-L. Choo, G. Kuo, R. Ralston, A. Weiner, D. Chien, G. Van Nest, J. Han, K. Berger, K. Thudium, J. Kansopon, J. McFarland, A. Tabrizi, K. Ching, B. Mass, L. B. Cummins, E. Muchmore, and M. Houghton, submitted for publication), suggesting that these purified HCV envelope proteins display native HCV epitopes. PMID- 8411379 TI - Mutations in the cytoplasmic tail of influenza A virus neuraminidase affect incorporation into virions. AB - The significance of the conserved cytoplasmic tail sequence of influenza A virus neuraminidase (NA) was analyzed by the recently developed reverse genetics technique (W. Luytjes, M. Krystal, M. Enami, J. D. Parvin, and P. Palese, Cell 59:1107-1113, 1989). A chimeric influenza virus A/WSN/33 NA containing the influenza B virus cytoplasmic tail rescued influenza A virus infectivity. The transfectant virus had less NA incorporated into virions than A/WSN/33, indicating that the cytoplasmic tail of influenza virus NA plays a role in incorporation of NA into virions. However, these results also suggest that the influenza A virus and influenza B virus cytoplasmic tail sequences share common features that lead to the production of infectious virus. Transfectant virus was obtained with all cytoplasmic tail mutants generated by site-directed mutagenesis of the influenza A virus tail, except for the mutant resulting from substitution of the conserved proline residue, presumably because of its contribution to the secondary structure of the tail. No virus was rescued when the cytoplasmic tail was deleted, indicating that the cytoplasmic tail is essential for production of the virus. The virulence of the transfectant viruses in mice was directly proportional to the amount of NA incorporated. The importance of the NA cytoplasmic tail in virus assembly and virulence has implications for use in developing antiviral strategies. PMID- 8411380 TI - Activation of the Epstein-Barr virus replicative cycle by human herpesvirus 6. AB - One common attribute of herpesviruses is the ability to establish latent, life long infections. The role of virus-virus interaction in viral reactivation between or among herpesviruses has not been studied. Preliminary experiments in our laboratory had indicated that infection of Epstein-Barr virus (EBV) genome positive human lymphoid cell lines with human herpesvirus 6 (HHV-6) results in EBV reactivation in these cells. To further our knowledge of this complex phenomenon, we investigated the effect of HHV-6 infection on expression of the viral lytic cycle proteins of EBV. Our results indicate that HHV-6 upregulates, by up to 10-fold, expression of the immediate-early Zebra antigen and the diffuse and restricted (85 kDa) early antigens (EA-D and EA-R, respectively) in both EBV producer and nonproducer cell lines (i.e., P3HR1, Akata, and Raji). Maximal EA-D induction was observed at 72 h post-HHV-6 infection. Furthermore, expression of late EBV gene products, namely, the viral capsid antigen (125 kDa) and viral membrane glycoprotein gp350, was also increased in EBV producer cells (P3HR1 and Akata) following infection by HHV-6. By using dual-color membrane immunofluorescence, it was found that most of the cells expressing viral membrane glycoprotein gp350 were also positive for HHV-6 antigens, suggesting a direct effect of HHV-6 replication on induction of the EBV replicative cycle. No expression of late EBV antigens was observed in Raji cells following infection by HHV-6, implying a lack of functional complementation between the deleted form of EBV found in Raji cells and the superinfecting HHV-6. The susceptibility of the cell lines to infection by HHV-6 correlated with increased expression of various EBV proteins in that B95-8 cells, which are not susceptible to HHV-6 infection, did not show an increase in expression of EBV antigens following treatment with HHV-6. Moreover, UV light-irradiated or heat-inactivated HHV-6 had no upregulating effect on the Zebra antigen or EA-D in Raji cells, indicating that infectious virus is required for the observed effects of HHV-6 on these EBV products. These results show that HHV-6, another lymphotropic human herpesvirus, can activate EBV replication and may thus contribute to the pathogenesis of EBV associated diseases. PMID- 8411381 TI - The replication functions of polyomavirus large tumor antigen are regulated by phosphorylation. AB - Polyomavirus (Py) large T antigen (T Ag) contains two clusters of phosphorylation sites within the amino-terminal half of the protein. To characterize possible regulatory effects of phosphorylation on viral DNA replication, Py T Ag was treated with calf intestinal alkaline phosphatase (CIAP). Incubation of the protein with a range of phosphatase concentrations caused progressive loss of phosphate without affecting its stability. Treatment with smaller quantities of CIAP stimulated the ability of the viral protein to mediate replication of constructs containing the viral replication origin, while higher concentrations of CIAP caused a marked diminution of this replication function. Several biochemical activities of Py T Ag were examined after CIAP treatment. Py T Ag DNA unwinding and nonspecific DNA binding were only slightly affected by dephosphorylation. However, as determined by DNase I footprinting experiments, treatment with smaller amounts of CIAP stimulated specific binding to the Py replication origin by Py T Ag, while treatment with larger amounts of CIAP caused marked inhibition of origin-specific binding by the viral protein. Phosphotryptic maps of Py T Ag before or after treatment with CIAP revealed changes in individual phosphopeptides that were uniquely associated with either the stimulation or the inhibition of replication. Our data therefore suggest that Py T Ag is regulated by both repressing and activating phosphates. PMID- 8411382 TI - Mutagenesis of the yellow fever virus NS2B protein: effects on proteolytic processing, NS2B-NS3 complex formation, and viral replication. AB - To study the role of specific regions of the yellow fever virus NS2B protein in proteolytic processing and association with the NS3 proteinase domain, a series of mutations were created in the hydrophobic regions and in a central conserved hydrophilic region proposed as a domain important for NS2B function. The effects of these mutations on cis cleavage at the 2B/3 cleavage site and on processing at other consensus cleavage sites for the NS3 proteinase in the nonstructural region were then characterized by cell-free translation and transient expression in BHK cells. Association between NS2B and the NS3 proteinase domain and the effects of mutations on complex formation were investigated by nondenaturing immunoprecipitation of these proteins expressed in infected cells, by cell-free translation, or by recombinant vaccinia viruses. Mutations within the hydrophobic regions had subtle effects on proteolytic processing, whereas mutations within the conserved domain dramatically reduced cleavage efficiency or abolished all cleavages. The conserved domain of NS2B is also implicated in formation of an NS2B-NS3 complex on the basis of the ability of mutations in this region to eliminate both association of these two proteins and trans-cleavage activity. In addition, mutations which either eliminated proteolytic processing or had no apparent effect on processing were found to abolish recovery of infectious virus following RNA transfection. These results suggest that the conserved region of NS2B is a domain essential for the function of the NS3 proteinase. Hydrophobic regions of NS2B whose structural integrity may not be essential for proteolytic processing may have additional functions during viral replication. PMID- 8411383 TI - Enhanced production of rat interleukin-8 by in vitro and in vivo infections with influenza A NWS virus. AB - We investigated the interleukin-8 (IL-8)-producing activity of influenza A NWS virus in cultured rat kidney NRK-52E cells and a rat influenza model. The production of rat IL-8 increased significantly in the virus-infected cells but not in UV-inactivated virus- or split-product-treated cells. The increase in IL-8 production could be detected in the bronchoalveolar lavage of infected rats. These data suggest that infectious virus has the potential to accelerate the production of IL-8 in cultured cells and in vivo in airway-lining cells. PMID- 8411384 TI - Expression of equine herpesvirus 1 glycoprotein D by using a recombinant baculovirus. AB - Glycoprotein D (gD) of equine herpesvirus 1 (EHV-1) was expressed at the surface of insect cells infected by a recombinant baculovirus. EHV-1 gD was detected as multiple forms (56, 52, and 48 kDa) from 18 to 96 h postinfection. Laboratory animals inoculated with the recombinant EHV-1 gD developed neutralizing antibody responses against both EHV-1 and EHV-4. PMID- 8411385 TI - Transcriptional trans activators of human and simian foamy viruses contain a small, highly conserved activation domain. AB - The Bel-1 protein of human foamy virus is a potent transcriptional trans activator of its homologous long terminal repeat promoter element. Here, we demonstrate that Bel-1 can also efficiently activate gene expression when targeted to a heterologous promoter by fusion to the DNA-binding motif of the yeast GAL4 protein. Analysis of a series of deletion mutants of Bel-1 generated in this hybrid protein context suggests the presence of a single transcription activation domain that is fully contained within a discrete, approximately 30 amino-acid segment located proximal to the Bel-1 carboxy terminus. Although this short motif can be shown to function effectively in eukaryotic cells of mammalian, avian, and fungal origin, it does not bear any evident sequence homology to the known classes of eukaryotic activation domain. However, this Bel 1 activation domain was found to be fully conserved, in terms of both biological activity and location, in the distantly related Taf trans activator of simian foamy virus type 1. PMID- 8411386 TI - Activation of a heterologous promoter by human immunodeficiency virus type 1 Tat requires Sp1 and is distinct from the mode of activation by acidic transcriptional activators. AB - We have previously shown that the Tat protein of the human immunodeficiency virus type 1 (HIV-1) is a modular transcriptional activator that can be targeted upstream of either a synthetic promoter or the intact HIV promoter to activate transcription. This activation was shown to be largely dependent on the presence of consensus binding sites for the cellular transcription factor Sp1. Since the use of heterologous promoters may provide further insight into Tat-mediated transactivation, we have analyzed the transactivation of the thymidine kinase promoter of herpes simplex virus by Tat and by the acidic transcriptional transactivator VP16. The effects of mutations of defined upstream promoter elements show that Tat transactivation is dependent on Sp1 binding sites in a site-specific manner. In contrast, transactivation by the acidic transactivator VP16 is completely independent of any of the defined promoter elements upstream of the TATA box. These results suggest that Tat and the classically defined modular acidic transcriptional activators have different modes of transactivation. In addition, the substitution of the HIV-1 TATA box for the thymidine kinase TATA box substantially increases Tat transactivation, indicating that Tat transactivation may also ultimately involve TATA box-associated cellular transcription factors. PMID- 8411387 TI - Infection by echoviruses 1 and 8 depends on the alpha 2 subunit of human VLA-2. AB - Anti-VLA-2 antibodies protected HeLa cells from infection by echoviruses 1 and 8 but not from infection by other echovirus serotypes. Echoviruses 1 and 8 bound to and infected nonpermissive hamster cells transfected with the alpha 2 subunit of human VLA-2. These results indicate that the human alpha 2 subunit is critical for infection by echoviruses 1 and 8 but that other echovirus serotypes must bind receptors other than VLA-2. PMID- 8411388 TI - Occurrence of alternatively spliced leader-delta onc-poly(A) transcripts in chicken neuroretina cells infected with Rous-associated virus type 1: implication in transduction of the c-mil/c-raf and c-Rmil/B-raf oncogenes. AB - We previously reported that serial passaging of Rous-associated virus type 1 in nondividing chicken embryo neuroretina cells leads to reproducible generation of acutely mitogenic retroviruses that transduced the catalytic domain of c-mil/c raf or c-Rmil/B-raf. On the basis of structural analysis of several retroviruses, we proposed that the early step of oncogene transduction is the constitution of alternatively spliced leader-delta onc-poly(A) transcripts. Here, we show that neuroretina cells do synthesize hybrid leader-delta mil and leader-delta Rmil RNAs and that these RNAs exhibit mitogenic properties and serve as templates for the generation of transducing retorviruses. PMID- 8411389 TI - Anoxia-inducible rat VL30 elements and their relationship to ras-containing sarcoma viruses. AB - VL30 elements are associated with cancer by their overexpression in rodent malignancies, their induction in a fibroblast response to anoxia which shares features with the malignant phenotype, and their presence recombined into Harvey murine sarcoma virus (HaSV) and Kirsten murine sarcoma virus. These sarcoma viruses contain ras oncogenes flanked on both sides by retrotransposon VL30 element sequences, in turn flanked by mouse leukemia virus sequences. Three very basic questions have existed about the VL30 element sequences found in sarcoma viruses: (i) how did they become recombined, (ii) what are their exact boundaries, and (iii) why are they there? To help decipher the nature of VL30 elements in sarcoma viruses, we examined VL30 clones isolated from an anoxic fibroblast cDNA library and independently by polymerase chain reaction cloning from rat cell DNA. Sequence comparisons with HaSV revealed that HaSV was formed by the substitution of 0.7 kb of VL30 sequences by 0.9 kb of c-Ha-ras sequences, with this event possibly facilitated by the presence of an identical Alu-like repeat found upstream of the 5' recombination point in both the VL30 element and c-Ha-ras. Recombination occurred 42 bases beyond the Alu-like sequences in VL30 and 1596 bases beyond them in c-Ha-ras, at position 926 of HaSV. The 3' ras-VL30 recombination event in HaSV occurred within a seven-base region of shared sequence identity, between HaSV bases 1825 and 1825 and 1831. Recombination between Moloney leukemia virus (MoLV) and VL30 appears to have occurred at a point corresponding to base 218 or 219 of MoLV and was near a TAR-like VL30 sequence; such recombination at the 3' end was between positions 7445 and 7456 of MoLV (HaSV positions 4694 to 4703). Kirsten murine sarcoma virus was found to be closely analogous to HaSV, and limited similar features were also seen with Rasheed sarcoma virus. PMID- 8411390 TI - Human immunodeficiency virus type 1 2-LTR circles reside in a nucleoprotein complex which is different from the preintegration complex. AB - The preintegration complex of human immunodeficiency virus type 1 (HIV-1) is a large nucleoprotein complex containing viral nucleic acids in association with products of the viral gag and pol genes. One of these proteins, integrase, is absolutely required for the integration and formation of the provirus. Although HIV-1-specific 2-LTR circles from nuclei of HIV-1-infected cells were found to be associated within a high-molecular-weight nucleoprotein complex, antibodies to HIV-1 integrase failed to precipitate this form of viral DNA. This result indicates that circular forms of HIV-1 DNA are not associated with integrase. These viral DNA forms seem to exist in a context of a nucleoprotein complex that is different from a preintegration complex of HIV-1. PMID- 8411391 TI - Molecular analysis of neurovirulent strains of Sindbis virus that evolve during persistent infection of scid mice. AB - To understand the role of tissue-specific adaptation and antibody-induced selectional pressures in the evolution of neurovirulent viruses, we analyzed three strains of Sindbis virus isolated from the brains of persistently infected scid mice and four strains of Sindbis virus isolated from the brains of scid mice with viral reactivation following immune serum treatment. For each viral isolate, we tested neurovirulence in weanling BALB/c mice and sequenced regions of the E2 and E1 envelope glycoprotein genes that are known to contain important determinants of Sindbis virus neurovirulence. One strain isolated from a persistently infected scid mouse and two strains isolated from scid mice with viral reactivation were neurovirulent, resulting in mortality in 80 to 100% of weanling BALB/c mice. All three neurovirulent strains contained an A-->U change at nucleotide 8795, which predicts a Gln-->His substitution at E2 amino acid position 55. No nucleotide changes were detected in the other sequenced regions of the E2 and E1 envelope glycoprotein genes or in the avirulent isolates. Our findings indicate that tissue-specific adaptations, rather than antibody-induced selectional pressures, are a critical determinant of the evolution of neurovirulent strains of Sindbis virus and provide evidence that E2 His-55 is an important neuroadaptive mutation that confers neurovirulence properties on Sindbis virus. PMID- 8411392 TI - Temperature-sensitive transforming mutants of the v-rel oncogene. AB - By making site-directed mutations in the avian retroviral oncogene v-rel, we created two temperature-sensitive (ts) transforming mutants; these changes were analogous to mutations previously shown to confer a ts function onto the Dorsal protein of Drosophila melanogaster. Chicken spleen cells infected with the ts v rel mutants formed colonies in agar at 36.5 degrees C but not at 41.5 degrees C. In addition, spleen cells derived from the ts v-rel-transformed colonies could be propagated in liquid culture at 36.5 degrees C but rapidly senesced at 41.5 degrees C. Both mutant v-Rel proteins were also ts for DNA binding in vitro. These mutants may be valuable for identifying genes directly regulated by v-rel. PMID- 8411393 TI - Inhibition of Rev activity and human immunodeficiency virus type 1 replication by antisense oligodeoxynucleotide phosphorothioate analogs directed against the Rev responsive element. AB - The interaction between the Rev protein of human immunodeficiency virus type 1 and its highly structured and conserved RNA target, the Rev-responsive element, is required for virus replication. We demonstrate that antisense oligodeoxynucleotide phosphorothioate analogs directed against the Rev-responsive element effectively inhibit Rev activity, as well as human immunodeficiency virus type 1 replication, and are candidates for antiviral therapy. PMID- 8411394 TI - Infection of macrophages with lymphotropic human immunodeficiency virus type 1 can be arrested after viral DNA synthesis. AB - Lymphotropic strains of human immunodeficiency virus type 1 (HIV-1), including HTLV-IIIB, replicate poorly in macrophages. We have shown previously that lymphotropic HIV-1 fuses equally well with T lymphocytes and macrophages (M. J. Potash, M. Zeira, Z.-B. Huang, T. Pearce, E. Eden, H. Gendelman, and D. J. Volsky, Virology 188:864-868, 1992), suggesting that events in the virus life cycle following virus-cell fusion limit virus replication. We report here that HIV-1 DNA is synthesized efficiently in either ADA or HTLV-IIIB infected alveolar macrophages or monocyte-derived macrophages within 24 h of virus infection, as observed by polymerase chain reaction for amplification of viral DNA sequences from the gag gene. Infection by a cloned lymphotropic HIV-1 strain, N1T-A, also leads to viral DNA synthesis. However, circular viral DNA was detected during strain ADA infection but not during HTLV-IIIB or N1T-A infection of monocyte derived macrophages. These findings indicate that during replication of lymphotropic HIV-1 in macrophages, all steps of the virus life cycle up to and including reverse transcription take place and that defects in later events, including DNA migration to the nucleus, may account for the limited production of viral proteins. PMID- 8411395 TI - Mutations in human immunodeficiency virus type 1 gp41 affect sensitivity to neutralization by gp120 antibodies. AB - Three closely related molecular human immunodeficiency virus type 1 (HIV-1) clones, with differential neutralization phenotypes, were generated by cloning of an NcoI-BamHI envelope (env) gene fragment (HXB2R nucleotide positions 5221 to 8021) into the full-length HXB2 molecular clone of HIV-1 IIIB. These env gene fragments, containing the complete gp120 coding region and a major part of gp41, were obtained from three different biological clones derived from a chimpanzee passaged HIV-1 IIIB isolate. Two of the viruses thus obtained (4.4 and 5.1) were strongly resistant to neutralization by infection-induced chimpanzee and human polyclonal antibodies and by HIV-1 IIIB V3-specific monoclonal antibodies and weakly resistant to soluble CD4 and a CD4-binding-site-specific monoclonal antibody. The third virus (6.8) was sensitive to neutralization by the same reagents. The V3 coding sequence and the gp120 amino acid residues important for the discontinuous neutralization epitope overlapping the CD4-binding site were completely conserved among the clones. However, the neutralization-resistant clones 4.4 and 5.1 differed from neutralization-sensitive clone 6.8 by two mutations in gp41. Exchange experiments confirmed that the 3' end of clone 6.8 (nucleotides 6806 to 8021; amino acids 346 to 752) conferred a neutralization sensitive phenotype to both of the neutralization-resistant clones 4.4 and 5.1. From our study, we conclude that mutations in the extracellular portion of gp41 may affect neutralization sensitivity to gp120 antibodies. PMID- 8411396 TI - Herpes simplex virus thymidine kinase and specific stages of latency in murine trigeminal ganglia. AB - From marker rescue, sequencing, transcript, and latency analyses of the thymidine kinase-negative herpes simplex virus mutant dlsactk and studies using the thymidine kinase inhibitor Ro 31-5140, we infer that the virus-encoded thymidine kinase is required in murine trigeminal ganglia for acute replication and lytic gene expression, for increasing the numbers of cells expressing latency associated transcripts, and for reactivation from latent infection. PMID- 8411397 TI - Context-dependent role of human immunodeficiency virus type 1 auxiliary genes in the establishment of chronic virus producers. AB - Two molecularly cloned viruses, human immunodeficiency virus type 1 (HIV-1)-NL4-3 (NL4-3) and HIV-1-HXB-2 (HXB-2), have been used to study the role of HIV-1 auxiliary genes in the establishment of chronic virus producers. NL4-3 encodes all known HIV-1 proteins, whereas HXB-2 is defective for three auxiliary genes: vpr, vpu, and nef. Studies were done in H9 cells, a T-cell line unusually permissive for the establishment of chronic virus producers. NL4-3 and HXB-2 undergo lytic phases of infection in H9 cultures with HXB-2, but not NL4-3, supporting the efficient establishment of chronic virus producers. Tests of mutant NL4-3 genomes containing various combinations of defective auxiliary genes revealed that both vpr and nef limited the ability of NL4-3 to establish chronic virus producers. Tests of a series of recombinants between NL4-3 and HXB-2 revealed that 5' internal sequences as well as fragments containing defective auxiliary genes affected the establishment of chronic virus producers. Viral envelope sequences and levels of virus production did not correlate with the ability to establish chronic virus producers. These results suggest that complex interactions of viral auxiliary and nonauxiliary gene functions with the host cell determine the ability to establish chronic virus producers. PMID- 8411398 TI - Generation of deletion mutants of simian immunodeficiency virus incapable of proviral integration. PMID- 8411399 TI - Recent advances in the diagnosis and treatment of renal arteriovenous malformations and fistulas. AB - Arteriovenous malformations remain relatively rare clinical lesions. However, with an increasing number of percutaneous renal procedures being performed, their overall incidence can be expected to increase. Congenital arteriovenous malformations usually present with hematuria, while acquired fistulas are more likely to present with hemodynamic changes, such as hypertension, cardiomegaly and congestive heart failure. Arteriography remains the principal method of diagnosis. However, newer imaging modalities such as CT, MRI and color duplex ultrasound may make significant contributions in the evaluation of these abnormalities (fig. 1). Surgery, whether nephrectomy or ligation of feeding vessels, has long been the standard treatment for symptomatic arteriovenous malformations or fistulas. The loss of normal renal parenchyma, migration of occluding agents or recanalization of abnormal vessels has limited the use of embolization as a method of treatment in the past. Recently, newer techniques and agents, such as pharmacoangiography and alcohol, have increased the efficacy of embolization therapy, either as an adjuvant to surgery, definitive therapy or palliation. Although to our knowledge there have been no controlled studies comparing surgical treatment versus embolization, certain recommendations can be made. Because of the decreasing morbidity and increasing efficacy, embolization should be attempted at the time of arteriography as a means of treatment for most arteriovenous malformations and fistulas. However, the choice of surgery, embolization or a combination must be individualized for each patient with regard to overall health, symptoms and manifestations of the fistula or malformation (fig. 2). PMID- 8411400 TI - Monitoring of emission as direct intraoperative control for nerve sparing retroperitoneal lymphadenectomy. AB - An intraoperative test to identify emission relevant lumbar postganglionic nerves during nerve sparing retroperitoneal lymph node dissection is presented. The neurophysiological course of the ejaculation into the posterior urethra, the emission, is described. A retroperitoneal nerve sparing procedure was done for nonseminomatous testicular tumors bilaterally in 7 patients with pathological stage I disease and unilaterally in 4 with pathological stage IIa disease. While the isolated lumbar nerves were electrostimulated, the seminal vesicles and bladder neck were monitored by suprapubic transvesical sonography. Simultaneously, endoscopy of the posterior urethra was performed and time code was registered. Emission began with bladder neck closure, propulsive contraction of the seminal vesicles in the periphery and opening of the paracollicular region. Then, complete contraction of the seminal vesicles was associated with closure of the prostatic urethra and ended in the separate secretion from the ductuli prostatici and ejaculatorii. Descending from nerve L1 to L3, their importance for emission usually increased. For intraoperative monitoring of emission transvesical sonography alone is sufficient. In 2 patients this method allowed us to identify the relevant nerves within the retroperitoneal residual mass (fibrosis) after chemotherapy. PMID- 8411401 TI - Evaluation of diagnostic tests in the differential diagnosis of primary aldosteronism: unilateral adenoma versus bilateral micronodular hyperplasia. AB - Effective management of primary aldosteronism is dependent upon correct localization of excessive aldosterone production. We report our results of localization studies in patients with biochemically and pathologically confirmed primary aldosteronism. Retrospective chart review identified 69 patients with unilateral adrenal adenoma and 11 with adrenal hyperplasia. Correct unilateral versus bilateral localization of excessive aldosterone production was predicted in 70% versus 71%, respectively, by adrenal venography, 100% versus 63% by adrenal vein hormone sampling, 46% versus 56% by adrenal nuclear scanning and 69% versus 13% by anomalous postural decline of aldosterone. Adrenal computerized tomography appeared to localize correctly 86% versus 80% of the lesions. Unilateral adrenalectomy normalized blood pressure in 79% of the patients with unilateral adenomas versus only 18% of those with adrenal hyperplasia. Once primary aldosteronism is confirmed, localization by adrenal vein sampling, adrenal venography and adrenal computerized tomography is most effective in directing antihypertensive therapy. PMID- 8411402 TI - Analysis of peripheral blood lymphocyte cell surface density of functional and activation associated markers in young and old hemodialysis patients. AB - Aging has been associated with specific shifts in various peripheral blood immune competent cell subsets. As part of pre-transplant immune profile evaluation possible parallel age-related changes in mean T-cell surface density of several cluster differentiation and activation linked antigens were into 2 groups: group 1-114 patients 40 years old or younger and group 2-36 patients 55 years old or older. Peripheral blood CD3+, DR+, CD3+DR+, CD4+, CD4+DR+, CD8+, CD8+DR+, CD56+, CD8+CD56+, CD3+IL-2-R+ and CD3+TR+ (interleukin-2 and transferrin receptors bearing CD3+ cells respectively), all mononuclear cells expressing IL-2-R and TR, and CD4+CD45+ cell subsets were analyzed and enumerated by 2-color flow cytometry. Subset relative levels as well as absolute counts were recorded. Cell surface density computation was performed using a computerized mathematical model based on fluorescence intensity vector analysis and cell size score determination based on light scatter pattern from raw data obtained by flow cytometry studies. Younger age was significantly associated with higher absolute cell count of CD3+ (p < 0.001), DR+ (p < 0.05), CD4+ (p < 0.01), CD8+ (p < 0.005), CD3+IL-2-R+ (p < 0.05), CD3+TR+ (p < 0.03) and IL-2-R+ (p < 0.05). Older patients had a slightly higher mean absolute count of CD4+CD45+ subset (p not significant) and significantly higher mean count for CD8+CD56+ cell subset (p < 0.001). When cell subset levels were compared between the 2 groups as the relative fraction of cells expressing a given marker out of all mononuclear cells gated out by flow cytometry, younger age was significantly associated with higher levels of CD3+ (p < 0.005), CD8+ (p < 0.001), CD4+DR+ (p < 0.004), CD3-TR+ (p < 0.05) and CD8+IL-2 R+ (p < 0.05). In contrast, slightly higher subset levels of CD56+ (p not significant), and significantly elevated levels of CD8+CD56+ (p < 0.0019) and CD4+CD45+ (p < 0.004) were observed in the older patients. Cell surface density analysis showed that younger patients had higher mean density per cell of CD3 (p < 0.05), CD8 (p < 0.001), IL-2-R on CD3+ cells (p < 0.05) and TR on CD3+ cells (p < 0.05). Mean cell surface density of CD56 on all CD56+ cells as well as on CD8+ cells was higher in older individuals (p < 0.001 and p < 0.003, respectively).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411403 TI - Routine use of indwelling ureteral stents in renal transplantation. AB - An extravesical ureteral implantation with the routine use of an internal stent was performed in 358 transplants (351 cadaveric and 7 living related). The 1-year patient and graft survival was 93% and 87%, respectively, with a minimum followup of 2 years. Ureteral complications developed in 9 patients (2.6%), with 3 fistulas, 2 of which resolved spontaneously, and 6 stenoses following stent removal. Nephrostomy drainage and antegrade stenting were initially attempted in all cases of stenosis, and were successful in 4. Revision of the ureteral anastomosis was required in 1 case of fistula and 2 cases of stenosis (0.9%). Extrinsic compression resulted in ureteral obstruction in 3 cases (2 lymphoceles and 1 hematoma), which resolved following drainage. Stent related complications occurred in 8 patients (2.2%), including obstruction due to the stent in 2 cases, breakage during removal in 3 leaving fragments in the upper urinary tract, proximal migration of 2 stents that were retrieved via percutaneous nephrostomy and calculus formation on 1 stent in a patient with hyperparathyroidism, necessitating extracorporeal shock wave lithotripsy for stent removal. In the cases with ureteral or stent related complications 1-year patient and graft survival was 100%. These results suggest that ureteral stents used routinely in renal transplantation are associated with a low incidence of urinary leaks, early postoperative obstruction and subsequent surgery for urological complications. However, a small number of unique problems related to stent use or malfunction may occur. Minimally invasive strategies using percutaneous nephrostomy and antegrade stenting are effective in managing the majority of complications that occur following ureteral stenting in renal transplant recipients. PMID- 8411404 TI - Renal oncocytoma: clinical and biological correlates. AB - We treated 31 patients with renal oncocytoma. Renal cell carcinoma was found existing separately within the same or contralateral kidney in 10 patients (32%). Followup of 29 patients revealed 24 alive with no evidence of disease and 1 alive with recurrent oncocytoma, while 1 with coexistent tumors died of progressive renal cell carcinoma and 3 tumor-free patients died of unrelated diseases. Quantitative deoxyribonucleic acid analysis was performed on cell suspensions of fresh tumor by flow cytometry or by image cytometry on touch preparations from frozen tissue in 16 patients with renal oncocytoma. Ploidy analysis revealed all oncocytomas to be diploid. Frozen tissue immunohistology was performed using murine monoclonal antibody against human HLA-A, B and C (class I) antigens with the avidin-biotin peroxidase technique in 11 patients with renal oncocytoma. Ten oncocytomas did not express these self-recognition antigens and 1 was only weakly positive for antigen expression. In contrast, renal cell carcinomas strongly expressed HLA class I antigens. The high incidence of coexistence of renal oncocytoma and renal cell carcinoma has important clinical implications. Loss of HLA class I antigen expression by renal oncocytomas may provide an additional method for differentiating this lesion from renal cell carcinoma. PMID- 8411405 TI - Interferon-alpha primed tumor-infiltrating lymphocytes combined with interleukin 2 and interferon-alpha as therapy for metastatic renal cell carcinoma. AB - Murine models demonstrate therapeutic synergy for the combination of interleukin 2, interferon-alpha and tumor-infiltrating lymphocytes. We treated 11 patients with metastatic renal cell carcinoma with a novel regimen consisting of in vivo primed tumor-infiltrating lymphocytes, interferon-alpha and interleukin-2. Patients received interferon-alpha before radical nephrectomy; in vivo primed tumor-infiltrating lymphocytes were isolated and expanded in vitro. Low dose continuous infusion interleukin-2 at a dose of 2 x 10(6) units per m.2 per day was administered for 96 hours during each treatment week and interferon-alpha was administered as a subcutaneous injection at a dose of 6 x 10(6) units per m.2 per day on days 1 and 4 of the interleukin-2 infusion. No therapy was given during the last 3 days of a treatment week. One course of therapy consisted of 3 weeks of therapy followed by 3 weeks of rest. Patients were treated until maximal response, disease progression or dose limiting toxicity. A maximum of 6 courses of therapy were administered. Eleven patients underwent interferon-alpha priming and subsequent radical nephrectomy. In vivo primed tumor-infiltrating lymphocytes were successfully expanded in all 11 patients with an expansion index of greater than 170. In vivo primed tumor-infiltrating lymphocytes maintained their lytic activity for greater than 5 to 8 weeks in culture as demonstrated in the 4-hour 51chromium release assay. Ten patients underwent multimodality biological therapy and 3 (30%, 95% confidence interval 6 to 65%) have achieved complete response (2 clinical and 1 surgical) with durations of 24+, 23+ and 5+ months. Patients with stable disease received no additional therapy. No deaths and no grade 4 toxicities occurred. Immunotherapy using a combination of interferon-alpha primed tumor-infiltrating lymphocytes, low dose continuous infusion interleukin-2 and interferon-alpha can induce significant and durable antitumor responses in some patients with advanced renal cell carcinoma. PMID- 8411406 TI - Does fish oil benefit stone formers? AB - The possibility that dietary fish oil supplementation may benefit patients with hypercalciuric urolithiasis by decreasing calcium excretion and enhancing protective mechanisms has been studied in rats and humans. In experiments on rats in metabolic cages, fish oil inhibited experimental nephrocalcinosis induced by intraperitoneal calcium gluconate. There were no significant changes in urinary biochemistry. In a clinical study on 18 hypercalciuric recurrent stone patients fish oil significantly decreased urinary calcium excretion. This effect was accompanied by decreases in the excretion of magnesium and citrate. Oxalate excretion and urinary fibrinolytic activity were unchanged. Overall, fish oil had a limited impact on the risk profile for recurrent urolithiasis. PMID- 8411407 TI - Ureteroscopic treatment of lower ureteral calculi in the era of extracorporeal shock wave lithotripsy: from a developing country point of view. AB - In a series of 229 lower ureteral calculi manipulated with either extracorporeal shock wave lithotripsy (ESWL) or ureteroscopy the stone clearance rate 7 days after treatment was 40.6% and 90%, respectively. Only 17 of 32 patients (53.1%) treated with ESWL were rendered stone-free at 4-week followup. In addition to concern about stone clearance, other medical and social factors, and the availability of health insurance coverage must be considered. Since the differences of living standards between highly developed and developing countries are great, the selection of ESWL or ureteroscopy as primary treatment of lower ureteral calculi must be based on the socioeconomical status of a specific country. Because ESWL devices are not readily available in most developing countries and ureteroscopy is cost-effective, highly efficacious (greater than 92%) and involves minimal morbidity (less than 10%) for calculi removal in experienced hands, we recommend ureteroscopy as the treatment of choice for lower ureteral stones, with ESWL as the alternative, in developing countries. PMID- 8411408 TI - The electromechanical impactor: the results of design modifications. AB - The first generation clinical electromechanical impactor was a 5F device that used a 3.0F electrohydraulic probe within a coiled spring with a blunt solder end cap. Clinical trials showed that this device fragmented 70% of the calculi but failed to fragment the harder stones. Applying the principle that impact kinetic energy is most dependent on velocity (KE = 1/2 MV2), 3 major design modifications were made to improve fragmentation efficiency: 1) the spring was changed from an extension to a compression type that captured and used more of each electrical spark, 2) the tip was changed from solder to lightweight titanium and 3) the tip shape was changed from blunt to conical. The result was a 41% increase in impact kinetic energy, a 40% increase in probe longevity and successful in vitro fragmentation of the harder calculi without compromise in tissue safety as determined by membrane perforation studies. PMID- 8411409 TI - Clinical experience with electromechanical impactor. AB - The electromechanical impactor is a 3.0F electrohydraulic electrode within a stainless steel sheath attached to a distal compression spring with a conical titanium tip. Each electrical discharge causes a 2.7 mm. tip extension and a kinetic impact energy of 900 bar. It is 5F, flexible and placed within the straight working port of a 9.5F to 10.0F rigid or semirigid ureteroscope. A clinical trial was performed and 15 patients (16 ureters with calculi) were treated with this device. An upper ureteral stone partially fragmented and migrated cephalad, 1 stone failed to break and was basket extracted, and there was 1 machine failure. The 13 other calculi were successfully broken to fragments less than 5 mm. In 2 patients baskets were also used to remove the larger monohydrate fragments. A secondary procedure was required to basket a 4 mm. calcium oxalate monohydrate fragment in a patient with 3 ureteral calculi. The average size of the calculi was 14 mm. in largest diameter. Excluding the machine failure, the average number of pulses required for fragmentation was 764. The life of each device was 600 to 800 pulses. Of the 15 cases 1 electrode was used in 10, 2 in 3 and 3 in 2. There was no evidence of ureteral wall injury, abrasion or perforation. The electromechanical impactor is a safe and effective method of ureteral stone fragmentation. PMID- 8411410 TI - Evaluation of urothelial Tamm-Horsfall protein and serum antibody as a potential diagnostic marker for interstitial cystitis. AB - Demonstration of adherence of Tamm-Horsfall protein to bladder epithelium has been suggested as a potential diagnostic test for interstitial cystitis. Bladder specimens from 18 interstitial cystitis patients were evaluated by indirect immunoperoxidase techniques using a Tamm-Horsfall protein specific monoclonal antibody to determine the diagnostic value of the staining results. The study population consisted of 7 severely diseased patients who required cystectomy with urinary diversion and 11 other patients meeting National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases criteria for interstitial cystitis. We were unable to detect intraepithelial or surface-bound Tamm-Horsfall protein in any of the biopsy tissues. Human kidney tissue, similarly fixed and processed, consistently demonstrated Tamm-Horsfall protein staining of the kidney tubules. The monoclonal antibody also reacted on Western blots against urinary Tamm Horsfall protein. Although antibody (alpha-Tamm-Horsfall protein) reactivity was measured by enzyme-linked immunosorbent assay in sera from interstitial cystitis patients, the titers did not differ statistically from those measured in sera from those without interstitial cystitis. Together, these results make it unlikely that immunohistochemical detection of Tamm-Horsfall protein will have diagnostic value in interstitial cystitis. Whether Tamm-Horsfall protein has a role in the pathogenic processes involved in this disease is not yet known. These findings do not eliminate the possibility that some interstitial cystitis patients will have abnormalities associated with the biochemical and physiological functions of Tamm-Horsfall protein. PMID- 8411411 TI - Invasive bladder cancer and uremia: dilemma in management. AB - Invasive bladder cancer presenting with uremia is a rare phenomenon. The management of these sick patients becomes problematic since one does not know how aggressive the treatment should be and what possible outcome to expect. In this retrospective review of 117 patients with invasive bladder cancer we describe 8 patients with uremia, and discuss our management strategy and prognosis. PMID- 8411412 TI - Success of the narrow-backed cuff design of the AMS800 artificial urinary sphincter: analysis of 144 patients. AB - The narrow-backed cuff design of the AMS800 artificial urinary sphincter was introduced to improve transmission of cuff pressure to underlying tissue and, theoretically, to decrease the incidence of cuff erosion. The AMS800 urinary sphincter with this design change was implanted in 136 male and 8 female patients (mean age 57 years). Mean followup was 28 months. The cuff was placed around the bladder neck in all 8 female and in 29 male patients, and it was implanted around the bulbous urethra in the remaining 107. The 4.5 cm. cuff in combination with the 61 to 70 cm. pressure balloon was implanted in 109 patients. There were 12 device failures and 12 surgical complications. Five patients required cuff removal and none required reoperation for inadequate cuff pressure. Postoperatively, 132 patients reported satisfactory continence. The design change of the AMS800 and the use of the 4.5 cm. cuff with the 61 to 70 cm. pressure balloon decreased the incidence of cuff erosion and the need for reoperation for inadequate cuff pressure. PMID- 8411413 TI - Repair of the complications of hypospadias surgery. AB - In 1992 a retrospective review was conducted of 190 patients evaluated and treated for complications of hypospadias surgery during 1979 through 1990 at the Devine Center for Genitourinary Reconstructive Surgery of Sentara Norfolk General Hospital and Children's Hospital of the King's Daughters. We could not contact 13 patients and 8 are awaiting a second stage procedure. Of the 177 patients 167 (94.35%) have had a successful outcome, defined as a controllable urinary stream, functional erection and an acceptable cosmetic appearance, and 2 (1.13%) are considered failures. Details of presenting problems, surgical techniques and recent modifications of these procedures are presented. PMID- 8411414 TI - Eversion of the tunica vaginalis for prophylaxis of testicular torsion recurrences. AB - After direct orchiopexy with punctiform suture fixation for prophylaxis of testicular torsion recurrence, at worst recurrent torsion may result in complete loss of the testis. Clinical and animal experience makes it appear more effective and reliable to prevent recurrent testicular torsion by eversion of the tunica vaginalis with extensive superficial adherence of the testicular integuments. We present the early and late results in 35 patients who underwent 46 eversion orchiopexies with a primary and definitive salvage rate of 87.5% and no case of recurrent testicular torsion. PMID- 8411415 TI - Management of chylous ascites after retroperitoneal lymph node dissection for testicular cancer. AB - Iatrogenic ascites is an uncommon complication of surgery of the retroperitoneum, the base of the mesentery or mediastinum. We treated 18 patients with chylous ascites occurring after retroperitoneal dissection for testicular cancer. Patients were diagnosed by paracentesis or on clinical grounds (increasing abdominal girth). Of interest, 6 patients underwent resection of the inferior vena cava as part of the procedure, and this appears to be a high risk group for this complication. Management options include dietary restriction of fat, administration of medium chain triglycerides and diuretics, hyperalimentation, peritoneovenous shunt or surgery. The majority of patients were managed successfully by dietary treatment. PMID- 8411416 TI - Morbidity with contemporary prostate biopsy. AB - To determine the incidence of complications associated with contemporary prostate biopsy, a review of 670 men undergoing transrectal prostate biopsy using 18 gauge biopsy needles was conducted. Of the men 580 received 1 to 3 days of ciprofloxacin antibiotic prophylaxis. A total of 16 patients (2.1%) suffered complications of whom 4 (0.6%) required hospitalization. These data demonstrate the low morbidity associated with contemporary transrectal prostate biopsy. PMID- 8411417 TI - Comparison of patient-controlled analgesia versus intramuscular narcotics in resolution of postoperative ileus after radical retropubic prostatectomy. AB - Patient-controlled analgesia has become standard practice after major abdominal operations. The benefits of patient-controlled analgesia have been well documented. However, its possible effect of prolonging postoperative ileus has not been well examined. To determine if patient-controlled analgesia prolongs postoperative ileus when compared to conventional intramuscular narcotics, a retrospective review of length of postoperative ileus in 98 consecutive patients (62 using patient-controlled analgesia and 36 using intramuscular narcotics) undergoing bilateral pelvic lymphadenectomy and radical retropubic prostatectomy was done. The patients receiving patient-controlled analgesia resolved the postoperative ileus an average of 1.0 day later than the intramuscular injection group (5.2 days versus 4.2 days p < 0.0001). Overall hospital stay was not significantly affected. Our results show that patient-controlled analgesia use prolongs postoperative ileus. PMID- 8411418 TI - The role of the diurnal variation of urinary pH in determining stone compositions. AB - The diurnal variation of urinary pH and urinary excretion of stone-forming constituents were investigated on 79 stone patients. Composition of the stones was analyzed by means of infrared spectrophotometry. In uric acid stone patients urinary pH was low during the 24-hour period. In mixed calcium oxalate-calcium phosphate stone patients urinary pH was low in the early morning but high in the daytime and at night. In magnesium ammonium phosphate stone patients urinary pH was high throughout the entire day. On the other hand, urinary excretion of calcium was high in the mixed calcium oxalate-calcium phosphate stone patients and low in uric acid or magnesium ammonium phosphate stone patients. These findings suggested that the diurnal variation of urinary pH and urinary excretion of calcium might have an important role in determining stone composition. PMID- 8411419 TI - Incision of the ureterovesical junction for endoscopic surveillance of transitional cell cancer of the upper urinary tract. AB - Ureteroscopy for treatment of upper tract transitional cell cancer is complicated by the need for subsequent outpatient, anesthesia-based, surveillance ureteroscopy to detect recurrent tumors. We report on 2 patients with ureteroscopically treated renal pelvic transitional cell carcinoma in whom the ureteral tunnel was incised. By rendering the ureterovesical junction incompetent, we created a widely patent refluxing ureteral orifice. During the last 2 years we have been able to perform repeatedly office-based flexible surveillance ureteroscopy without fluoroscopy and without oral or parenteral analgesics in both patients. PMID- 8411420 TI - Urodynamic evaluation of results of endopyelotomy for ureteropelvic junction obstruction. AB - We treated 13 patients with unilateral ureteropelvic junction obstruction by percutaneous endopyelotomy, and they were urodynamically evaluated by the Whitaker test and diuresis renography, in addition to excretory urography (IVP). Surgery was uncomplicated in all patients except 1 who required repeat incision 4 weeks later. Obstruction was diagnosed preoperatively by the Whitaker test and diuresis renography in 10 cases, and by IVP in 13. Postoperatively, all 10 patients (100%) with a positive Whitaker test were free of obstruction with a significant reduction in the relative renal pelvic pressure from 35.0 to 8.1 cm. water (p < 0.01). Diuresis renography revealed no obstruction in 8 patients (80%) and persistent obstruction in 2. IVP demonstrated reduced hydronephrosis in 8 of 13 patients (62%) 12 weeks after surgery and in 11 of 13 patients (85%) 19 months later. However, a marked reduction in renal pelvic size was noted in only 3 patients (23%). Flank pain disappeared in 10 of 11 patients (91%) postoperatively. Overall, surgery was successful in 11 of the 13 patients (85%). Percutaneous endopyelotomy was effective in relieving obstruction at the ureteropelvic junction with minor morbidity. The Whitaker test was more sensitive for evaluating the results of surgery than diuresis renography and IVP, although diuresis renography appears to be useful in followup evaluation of hydronephrosis. PMID- 8411421 TI - Management of urethral obstruction with transvaginal urethrolysis. AB - Transvaginal urethrolysis was performed in 48 patients with urethral obstruction following 1 or more urethral suspension procedures. A needle suspension procedure had been performed in 19 patients (40%), retropubic urethropexy in 17 (35%) and pubovaginal sling in 10 (21%). A good response as indicated by normal voiding with minimal or no irritative symptoms was achieved in 33 patients (65%). The most common presenting complaints were irritative voiding symptoms (71%) and urinary retention (60%). Fluoroscopic urodynamic evaluation frequently demonstrated urethral hypersuspension, a cystocele and/or elevated voiding pressures. No patient had stress urinary incontinence as a result of the procedure. Urethral obstruction should be recognized as a potential complication following surgical correction of female stress urinary incontinence. Transvaginal urethrolysis is a safe and effective method to manage this problem. PMID- 8411422 TI - Clinical assessment of urethral sphincter function. AB - Measurements of urethral pressures, such as maximum urethral pressure, are widely believed to have relevance in the management of urinary incontinence despite evidence to the contrary. In this study maximum urethral pressure and the abdominal pressure required to cause stress incontinence were measured in 125 women with stress incontinence. In women the abdominal pressure required to cause stress incontinence was unrelated to maximum urethral pressure. These findings indicate that maximum urethral pressure has little relationship to urethral resistance to abdominal pressure. In the 9 children with myelodysplasia we compared the detrusor pressure with the abdominal pressure required to induce urethral leakage. These values also were quite different, indicating that as far as the urethra is concerned abdominal pressure and detrusor pressure are not equivalent forces. PMID- 8411423 TI - Incontinence. PMID- 8411424 TI - Management of synchronous bilateral Wilms tumor: Brazilian Wilms Tumor Study Group experience with 14 cases. Brazilian Wilms Tumor Cooperative Group, Sao Paulo, Brazil. AB - The Brazilian Wilms Tumor Study Group has registered 16 cases of synchronous bilateral Wilms tumor since 1986, of which 14 were available for analysis. Most patients were treated according to a prospective trial in which bilateral renal preservation was emphasized. Overall survival was 79% (11 of 14 cases), and both kidneys were preserved in 57% (8 of 14) of the patients with synchronous disease. Of the 14 patients 5 had undergone at least 3 surgical procedures. In 10 cases in which pathological reports were available for review residual disease was documented in 5 but none had relapse. PMID- 8411425 TI - Ureteral reimplantation in infants. AB - Between 1984 and 1990, 30 infants (46 ureters) 8 weeks to 6 months old (mean age 4 months) underwent ureteral reimplantation. Weight at operation ranged from 4.9 to 9.5 kg. (mean 6.9). Underlying abnormalities were primary vesicoureteral reflux (28 ureters), primary ureterovesical junction obstruction (11), ectopic ureterocele (4) and ectopic ureter (3). Patients with reflux underwent surgery because of high grade reflux (grade IV or V) or breakthrough infection. Infants with primary ureterovesical junction obstruction underwent obstructive diethylenetriaminepentaacetic acid diuretic renograms. Reimplantations performed included 44 Glenn-Anderson advancements, 1 Cohen cross-trigonal advancement and 1 Politano-Leadbetter procedure. Of the ureters 30 (65%) were tapered intravesically. Ureteral stents were used in all instances. Transient ureteral obstruction developed in 2 children following stent removal and 1 required temporary percutaneous nephrostomy drainage. No permanent ureterovesical obstruction was noted in any patient. Followup at 18 months revealed no postoperative reflux in 43 of 46 ureters (93%). One infant required repeat reimplantation to correct a vesicoureteral fistula and the remaining 2 patients (2 ureters) with low grade reflux (grade I and II) are being observed. Surgery was successful in 27 of 30 tapered ureters (90%) and in all 16 of the nontapered ureters (100%). While the majority of infants with ureterovesical junction abnormalities may be observed, some may require surgery. Reimplantation, when necessary in this age group, can be performed with a high degree of success and diverting procedures such as vesicostomy or ureterostomy can be avoided. PMID- 8411426 TI - Enterocystoplasty in renal transplantation candidates: urodynamic evaluation and outcome. AB - In anticipation of renal transplantation 11 patients with end stage renal disease and small volume, poorly compliant bladders underwent enterocystoplasty. The etiology of bladder dysfunction included myelodysplasia in 4 patients, posterior urethral valves in 4 and vesicoureteral reflux in 3. Mean patient age at the time of bladder augmentation was 16.4 years (range 10 to 28). Detubularized bowel segments were used in all patients and included ileum in 7, ileocecal segment in 3 and sigmoid in 1. Urodynamic evaluation before and after enterocystoplasty demonstrated marked improvement in bladder capacity and compliance. These 11 patients have received 13 renal transplants with an 85% graft survival at a mean followup of 30.1 months. No bladder ruptures or other perioperative complications have occurred. All patients remain continent on clean intermittent catheterization. Our experience suggests that enterocystoplasty is a safe and effective method of restoring lower urinary tract function in the patient with end stage renal disease and a small, noncompliant bladder. PMID- 8411427 TI - Intravesical urokinase for the management of clot retention in boys. AB - We successfully used intravesical urokinase in 2 young boys to relieve clot retention occurring as a consequence of upper tract bleeding. Urokinase clot dissolution appears to offer a simple, minimally invasive, atraumatic solution to this infrequent problem. PMID- 8411428 TI - A case of primary malignant lymphoma of the bladder in childhood. AB - Primary malignant lymphoma of the bladder is a rare entity. Review of the reported cases of bladder lymphoma in the literature reveals that bladder lymphoma is typically found in elderly patients. We report a well documented case of a non-Hodgkin's primary bladder lymphoma in childhood. The unusual clinical presentation, immunochemical analysis, treatment and 5-year followup are discussed. PMID- 8411429 TI - Carcinoma of the urachus in an adolescent: a case report. PMID- 8411430 TI - Posterior hypospadias: long-term followup after reconstructive surgery in the male direction. AB - Posterior hypospadias with a scrotal or perineal meatus results from a defect in step 3 of male sexual differentiation. The different etiological factors underlying this condition result in a broad spectrum of presentations ranging from the isolated form to complex ambiguity of the external genitalia, such as male pseudohermaphroditism. Between 1952 and 1988 a total of 92 patients with posterior hypospadias underwent a 2-stage reconstruction at our department. A retrospective study was performed with the aim of evaluating the long-term results in these patients. Our special interest focused on the functional and cosmetic results, exocrine and endocrine functions, as well as the sexual lives of the patients. While satisfactory results were obtained in two-thirds of the 42 male patients available for long-term followup, there were 13 patients who at followup still presented with complex sexual ambiguity. In 6 of these patients androgen receptor defects were detected by means of biochemical as well as molecular-biological investigations. Our data emphasize the importance of androgen metabolism for male sexual development and underline the necessity of careful evaluation in these children. PMID- 8411431 TI - The incidence of congenital penile curvature. AB - To determine the incidence of congenital penile curvature a group of 500 consecutive male neonates at our institution were examined for this anomaly. In this group there were 3 cases of congenital penile curvature, for an incidence of 0.6%. PMID- 8411432 TI - Association of sickle cell disease, priapism, exchange transfusion and neurological events: ASPEN syndrome. AB - Priapism and acute neurological events are believed to be unrelated complications of sickle cell hemoglobinopathy. We describe a syndrome based on our experience and a review of the literature of significant neurological events after partial exchange transfusion to treat priapism in sicklemic patients. Severe headache is often the initiating symptom of this complex. The ensuing neurological events range from seizure activity to obtundation requiring ventilatory support. The proposed pathophysiology of these neurological events is related to cerebral ischemia after an acute increase in per cent total hemoglobin, concomitant decrease in per cent hemoglobin S and subsequent release of vasoactive substances during penile detumescence. We have termed this constellation of events the ASPEN syndrome, an eponym for association of sickle cell disease, priapism, exchange transfusion and neurological events. Early recognition and aggressive medical management resulted in complete reversal of neurological sequela. PMID- 8411433 TI - Laparoscopic marsupialization of a simple renal cyst. AB - An 8 cm. symptomatic simple renal cyst, recurrent despite previous aspiration and injection of sclerosing agents, was marsupialized using laparoscopic techniques. No complications occurred and followup by ultrasonography confirmed resolution of the lesion. We suggest that laparoscopic management of symptomatic simple renal cysts may be an attractive alternative to open surgical techniques. PMID- 8411434 TI - Fatal pulmonary embolus from ischemic necrosis of intracaval tumor thrombus: a case report. AB - Angioinfarction of renal tumors has been proposed as a preoperative adjunct and as palliative therapy. Most side effects of angioinfarction are transient and are well tolerated by the patient. We report a case of fatal pulmonary embolus resulting from migration of an intracaval tumor thrombus following renal arterial embolization. PMID- 8411435 TI - Extra-adrenal perirenal myelolipoma. AB - Extra-adrenal perirenal myelolipomas are rare benign tumors that consist of adipose and hematopoietic tissue. We report a case of perirenal myelolipoma in an otherwise healthy man and review the literature. PMID- 8411436 TI - Corticosteroid-associated fatal mycobacterial sepsis occurring 3 years after instillation of intravesical bacillus Calmette-Guerin. AB - A case is reported of systemic Mycobacterium bovis infection that occurred 3 years after uneventful instillation of intravesical bacillus Calmette-Guerin (BCG) and after several months of oral prednisone therapy. The literature on delayed BCG infection and the systemic persistence of BCG after intravesical instillation is reviewed. We propose that rarely a reservoir of dormant M. bovis may become established after intravesical therapy. Reactivation infection may later develop in a manner that parallels the natural history of secondary tuberculosis. PMID- 8411437 TI - Scorpion sting on the penis. AB - A 58-year-old man was stung on the penis by a scorpion of the species Tityus serrulatus. The patient felt a severe sharp local pain that did not improve until he presented to the hospital 12 hours later. Block of the dorsal nerves of the penis with 1% lidocaine led to immediate relief of the pain. The aim of treatment of a Tityus scorpion sting on the penis, without systemic manifestations, should be concentrated on relief of the pain. Local or regional block with a local anesthetic is the treatment of choice. The use of the specific antivenom is recommended only for cases of systemic manifestations, particularly in children. PMID- 8411438 TI - Treatment of corpus cavernosum abscess by aspiration and intravenous antibiotics. AB - We report the successful treatment of an abscess in the corpus cavernosum of a paraplegic man on intracavernous papaverine injection therapy by means of percutaneous aspiration and intravenous antibiotics. After 3 months of intracavernous pharmacotherapy the patient experienced good erections again and only minimal fibrosis could be found on palpation. PMID- 8411439 TI - Chronic priapism secondary to a traumatic arteriovenous fistula of the corpus cavernosum. AB - A 30-year-old man presented with a 7-month history of chronic partial priapism and sexual dysfunction following blunt perineal trauma. Duplex penile ultrasound confirmed high flow priapism and color flow Doppler ultrasound diagnosed an arteriovenous fistula within the left corpus cavernosum. This lesion was corrected via corporeal exploration and ligation of the artery feeding the fistula. The use of color flow Doppler ultrasound to diagnose this unusual condition obviated the need for angiographic studies. This rare condition of posttraumatic priapism may be diagnosed by noninvasive means. The pathophysiology, diagnosis and treatment of posttraumatic priapism and review of the literature are discussed. PMID- 8411440 TI - A case of testicle replantation. AB - A case is reported of amputation of the testis, which was reattached to the stump of the severed spermatic cord using a microsurgical technique and local hypothermia. Total ischemia time was 4 hours and 20 minutes, including 3 hours for warm ischemia. Ultrasonography of the replanted testis 6 months postoperatively showed a homogeneous parenchymal echo pattern and color-coded duplex sonography confirmed normal blood flow. Semen analysis and serum testosterone level were normal. A biopsy of the replanted testicle revealed active spermatogenesis and a normal number of Leydig cells. PMID- 8411441 TI - Familial occurrence of Leydig cell tumors: a report of a case in a father and his adult son. AB - We report on the familial occurrence of nonhormone-secreting Leydig cell tumors in a father and his adult son. No evidence for an underlying endocrinological disorder was found. The father even had metachronous bilateral Leydig cell tumors with an interval of 11 months. PMID- 8411442 TI - Carney's complex in a patient with hormone-producing Sertoli cell tumor of the testicle. AB - Carney's complex is an unusual disorder consisting of a variety of endocrinological and urological manifestations. The characteristic gonadal and adrenal features of Carney's complex should become familiar to urologists. A patient was evaluated for obesity, cushingoid features, hyperlipidemia, glucose intolerance, coronary artery disease, a left adrenal mass, bilateral testicular masses and cardiac myxomas. Pathological evaluation revealed the testicular tumors to be of Sertoli cell origin, the adrenal mass to be an adrenocortical adenoma and intracardiac lesions consistent with myxomas. The features of Carney's complex are discussed. PMID- 8411443 TI - Sonographic appearance of left spermatic vein thrombosis simulating incarcerated inguinal hernia. AB - We report a case of exercise-induced spontaneous thrombosis of the left spermatic vein. The sonographic appearance mimicked an incarcerated left inguinal hernia by demonstrating a tubular, hypoechoic, noncompressible, cystic appearing inguinal mass with no flow evident on color Doppler imaging. PMID- 8411444 TI - Small bowel obstruction following laparoscopic lymphadenectomy. AB - We report 2 cases of small bowel obstruction following staging laparoscopic pelvic lymph node dissection for prostate adenocarcinoma. Short segments of ileum became incarcerated in the right lower abdominal trocar sites (10 mm.) in both patients. Laparotomy was done to repair the problem in both cases and in 1 small bowel resection was required because of strangulation. This complication can probably be prevented in the majority of cases by meticulous attention to trocar site closure at the termination of a laparoscopic procedure. PMID- 8411445 TI - Urogenital malacoplakia treated with fluoroquinolones. AB - We report 2 cases of urogenital malacoplakia and discuss the problems associated with diagnosing and treating this rare disease. In 1 case extensive pelvic disease was successfully controlled with ciprofloxacin. In the other case the malacoplakia was localized in the distal ureter and was initially mistaken for urothelial carcinoma. Nephrectomy was performed due to persistent ureteral obstruction and renal dysfunction. In the management of urogenital malacoplakia it seems essential that urinary tract infections be effectively treated. The new fluoroquinolones appear to be potentially successful in managing this disorder due to their high uptake of macrophages. PMID- 8411446 TI - Topical effect of intravesical oxybutynin. AB - Intravesical oxybutynin was instilled into rat bladders in graded doses by repeated catheterization to study the local bladder effects of drug concentration and incidental urinary infection. On the thirteenth day, after 5 doses, bladders were recovered for measurement and histological study. In the highest dose group, systemic absorption from the bladder caused weight loss and cachexia. However, in no group was there any clear evidence of drug-related mucosal or bladder wall change. Instead, the two high dose groups seemed somehow protected from the combined adverse effects of catheterization and infection. PMID- 8411447 TI - Combined antitumor effect of suramin plus irradiation in human prostate cancer cells: the role of apoptosis. AB - Suramin has recently surfaced as a potential antineoplastic agent on the basis of its ability to exert a cytostatic effect on human prostate carcinoma cells. Radiotherapy for the treatment of prostate cancer has long been known as an alternative medical therapeutic approach, but the molecular mechanism involved in radiation-induced toxicity in prostatic tumors is poorly defined. In these studies, the antitumor effect of suramin and irradiation, either as individual treatments or in combination, was investigated in human prostate cancer cells. Two androgen-independent prostate cancer cell lines, DU-145 and PC-3, were used as in vitro model systems to study the underlying molecular mechanisms of these two therapeutic modalities. A cytostatic effect on cell growth was observed when cells were exposed to suramin alone, while treatment with irradiation alone resulted in significant cell death as determined by the Trypan blue exclusion assay. Suramin treatment prior to irradiation inhibited this radiation-induced cell death. In contrast, exposure of cells to suramin following irradiation enhanced the cytotoxic effect of ionizing radiation. Temporal analysis of the molecular events involved in radiation-induced toxicity revealed the characteristic fragmentation of DNA into a nucleosomal ladder (a hallmark of apoptosis) and enhanced expression of specific programmed cell death-associated genes (TRPM-2 and TGF-beta), preceding the dramatic decrease in cell number. These results indicate that radiation-induced cell death proceeds via the apoptotic pathway. Further studies have demonstrated that activation of programmed cell death by ionizing radiation is substantially inhibited by pretreatment of the cells with suramin. This study suggests that the relative timing of this combination treatment may have significant therapeutic implications in the treatment of advanced prostate cancer. PMID- 8411448 TI - The influence of acute overdistension on rat bladder function and DNA synthesis. AB - Prolonged micturition problems are often encountered after long-term bladder overdistension caused by urinary retention. In animal studies, damage to the bladder wall innervation has been found following overdistension. Experimentally, acute overdistension has also been implicated in the pathogenesis of the response to partial outlet obstruction. In the present study we investigated the influence of overdistension on micturition volume and frequency, on in vitro bladder function using the whole bladder model and on 3H-thymidine uptake, localization and DNA synthesis. Overdistension was induced for 3 hours by forced diuresis and balloon obstruction. Another group of rats was catheterized for 3 hours but received no diuretic, nor was the balloon inflated. An additional group of controls was neither anesthetized nor catheterized. Overdistension caused a gradual increase in bladder mass which was maximal at 7 days. During the first 24 hours following overdistension, the frequency of micturition decreased, but normalized thereafter. A progressive decrease in the response to field stimulation was noted between 16 hours and 7 days following overdistension and remained at this level until 21 days. There were, however, no significant differences in the responses to carbachol, ATP and KCl. There was a 30% reduction in the ability of field stimulation to empty the bladder 16 hours after overdistension, but no impairment of the emptying ability of carbachol. Overdistension was followed by a significant increase in 3H-thymidine uptake, which was maximal at 2 days. 3H-thymidine labelling increased rapidly after overdistension and was maximal within 16 hours in the urothelium. In smooth muscle, connective tissue and lamina propria, maximal labelling occurred at 2 days. Catheterization alone caused a mild distension which was associated with a small, but statistically significant, increase in 3H-thymidine incorporation into DNA within 16 hours. The labelling was located primarily in the urothelium. Overdistension causes a proliferative reaction within the bladder wall. Its initial effects occur within the urothelium, and the later involvement of the subendothelial smooth muscle and connective tissue is directly proportional to the degree of bladder distension. Three weeks following overdistension, the bladder's functional state was not completely recovered, although the urinary bladder was found to have a good capacity to adapt and compensate for the stress induced changes caused by overdistension. It is, therefore, clear that overdistension may have long-lasting effects on the bladder. PMID- 8411449 TI - Effect and content of arginine vasopressin in normal and obstructed rat urinary bladder: an in vivo and in vitro investigation. AB - The present investigation was performed to evaluate the possible role of arginine vasopressin (AVP) in detrusor instability in the obstructed rat urinary bladder. The effect of AVP on normal and obstructed rat detrusor smooth muscle was tested in vivo and in vitro. Arginine vasopressin given as a closed intraarterial injection to the bladder transiently decreased micturition volume and increased micturition frequency during cystometry in control rats. In rats with infravesical outlet obstruction the effect of AVP on cystometrical parameters was negligible. In accordance with this finding, the efficacy of AVP in contracting detrusor muscle in vitro was much lower for obstructed bladders than for controls. The EC50 values were, however, similar. Arginine vasopressin added to the bath had no effect on nerve-mediated contractile responses. Obstruction led to a transient decrease in immunoreactive AVP concentration, but the total amount of AVP per bladder increased significantly after 6 weeks of obstruction as a consequence of the 14-fold increase in bladder weight. The decreased excitatory effect of AVP in obstructed bladder makes a role for this peptide in the development of detrusor instability less likely. PMID- 8411450 TI - Comparison of MGH-1 and TuMark antibody detection in transitional cell carcinoma. AB - Two monoclonal antibodies, TuMark-BTA and MGH-1, were evaluated in patients with benign pathology, history of transitional cell carcinoma with no active disease and active transitional cell carcinoma. Both antibodies were accurate in predicting carcinoma in approximately two-thirds of patients. TuMark showed a 74% sensitivity and 50% specificity, while MGH-1 was more sensitive (86%) and less specific (38%). Positive predictive values for TuMark and MGH-1 were 79% and 78%, respectively. PMID- 8411451 TI - The effect of indomethacin infusion on renal hemodynamics and on the renin angiotensin system during unilateral ureteral obstruction of the pig. AB - Obstruction of the urinary tract is associated with an increase in pelvic pressure (PP) and a decline in ipsilateral renal blood flow (RBF). To investigate the influence of the renal prostaglandins on these parameters, pigs with complete unilateral ureteral obstruction (UUO) were studied under general anesthesia after administration of indomethacin 30 minutes before obstruction. Pelvic pressure increased to a maximum of 41.1 +/- 2.4 cm. H2O after 6 hours, but maximum pressure, as well as the rate of pressure increase, was significantly reduced after indomethacin. A transient but significant increase in mean aortic blood pressure was seen, together with a bilateral decrease in RBF immediately after indomethacin administration. Ipsilaterally RBF decreased by 25% from 319 +/- 21 ml. per minute to 237 +/- 16 ml. per minute and contralaterally by 23% from 300 +/- 20 ml. per minute to 231 +/- 25 ml. per minute. After 15 hours a further decline was measured in ipsilateral RBF to 136 +/- 20 ml. per minute but, on the contralateral side, RBF was only slightly reduced to 281 +/- 36 ml. per minute after 15 hours. Following indomethacin a sustained increase in ipsilateral renal vascular resistance (RVR) was observed from 37.6 +/- 3.6 mm. Hg x ml.-1 x min. x gm. to 130.5 +/- 42.3 mm. Hg x ml.-1 x min. x gm. compared with a much smaller increase from 37.4 +/- 3.2 mm. Hg x ml.-1 x min. x gm. to 55.1 +/- 9.2 mm. Hg x ml.-1 x min. x gm. in the control group. Renal secretion rate of angiotensin II was significantly reduced 2 and 6 hours after indomethacin administration on the obstructed side, and was not significantly changed on the contralateral side. It can be concluded that inhibition of prostaglandin synthesis during unilateral ureteral obstruction in pigs results in reduced pelvic pressure, renal blood flow and angiotensin II secretion rate from the affected kidney, and that renal prostaglandin synthesis plays an important role for the perfusion of the kidney during ureteral obstruction. PMID- 8411452 TI - The fetal rat gubernaculum contains higher levels of androgen receptor than does the postnatal gubernaculum. AB - The androgen receptor of the rat gubernaculum was measured by a sensitive immunoblotting technique from day 19 of fetal development to day 20 of postnatal development. In relative terms (densitometric units/microgram. protein), it was found that the amount of the gubernacular androgen receptor decreased dramatically from fetal to postnatal development, coincident with the transition of the gubernaculum from a tissue primarily composed of undifferentiated mesenchymal cells in the fetus to a tissue that is primarily made up of muscle during postnatal development. We conclude that the undifferentiated mesenchyme of the fetal gubernaculum is a primary target of androgen action. PMID- 8411453 TI - Structural basis of geriatric voiding dysfunction. I. Methods of a prospective ultrastructural/urodynamic study and an overview of the findings. AB - Voiding dysfunctions are common in the elderly. Yet the pathogenesis and pathophysiology have remained largely unknown. To date there has been little information on structure of the aging detrusor. To gain insight into the structural basis of voiding dysfunction in the elderly, we examined detrusor biopsy specimens by electron microscopy. The specimens were obtained from 24 women and 11 men 65 to 96 years old (mean age 79 years) who were carefully selected by detailed clinical and neurological examination. Symptom-free subjects were particularly sought to identify those who might provide the structural/functional norm of aging detrusor. Comprehensive urodynamic study was performed in all subjects. A transurethral detrusor biopsy was obtained and processed to study ultrastructure of the smooth muscle, intrinsic nerves and interstitium. Subjects were segregated purely by urodynamic findings, regardless of symptoms, into detrusor overactivity, outlet obstruction, obstruction plus overactivity and neither (that is no obstruction and no overactivity) groups, each with a subgroup of normal and another of impaired contractility. Specimens were segregated blindly and independently by ultrastructural features into dysjunction, myohypertrophy, myohypertrophy plus dysjunction and dense band patterns, each with a subset with widespread degeneration of muscle cells and nerves, and another with minimal or no degeneration. When codes were broken, each structural pattern (and subset) matched with a specific urodynamic group (and subgroup)--with no overlap. The dysjunction pattern matched with overactivity, the myohypertrophy pattern with obstruction, the myohypertrophy plus dysjunction pattern with obstruction plus overactivity, and the dense band pattern with the neither group. Structural subsets of widespread degeneration matched with impaired contractility subgroups, and subsets with minimal or no degeneration matched with normal contractility subgroups. These observations identify specific structural bases of the major forms of geriatric voiding dysfunction, provide important insights into their pathogenesis, and introduce detrusor biopsy as a potentially valuable tool in their diagnosis and clinical management. PMID- 8411454 TI - Structural basis of geriatric voiding dysfunction. II. Aging detrusor: normal versus impaired contractility. AB - Little information on the structural norm of the aging detrusor is currently available. To gain insight into the pathophysiology of geriatric voiding dysfunction, detrusor biopsies were examined by electron microscopy to identify structural correlates of specific, urodynamically defined abnormalities of vesical function in 35 elderly subjects. Prospective urodynamic grouping of the subjects and segregation of the detrusor specimens by ultrastructural features were done independently and blindly. One structural pattern so identified, the dense band pattern, matched the urodynamic group with neither detrusor overactivity nor bladder outlet obstruction. This neither group included 11 women and 2 men 65 to 91 years old (mean age 76 years). Except for 2 patients with minimal stress incontinence, all were symptom-free. None of the patients had diabetes or a neurological deficit. Urodynamically, 10 patients had impaired and 3 had normal detrusor contractility. The dense band structural pattern was characterized by overall normal configuration of muscle cells and cell junctions, sarcolemma (muscle cell membrane) dominated by dense bands with depleted caveolae in interposed zones and slight widening of spaces between muscle cells with little-collagen content. Specimens from the 10 subjects with impaired contractility displayed, in addition, widespread degeneration of muscle cells and axons. The remaining 3 specimens, without degeneration, matched the subjects with normal contractility, who were continent and symptom-free. It is proposed that the dense band pattern represents the structural norm of aging detrusor, heralds a process of muscle cell de-differentiation in the detrusor accompanying natural aging, and may affect exchange and storage of ions involved in the excitation contraction coupling mechanism of muscle cells through depletion of caveolae. Widespread degeneration of muscle cells and axons, superimposed on the dense band pattern, is proposed as the structural correlate of impaired detrusor contractility in the aging detrusor. PMID- 8411455 TI - Structural basis of geriatric voiding dysfunction. III. Detrusor overactivity. AB - Detrusor overactivity in the absence of outlet obstruction is common in the elderly. The few available studies on structure of the overactive detrusor generally have dealt only with its innervation. We conducted a prospective study to examine the ultrastructure of muscle cells, interstitium and nerves of the detrusor in biopsies from 35 elderly subjects to identify structural correlates of various urodynamically defined forms of voiding dysfunction. A distinctive dysjunction structural pattern was identified blindly in 15 detrusor biopsies. These patterns matched 12 women and 3 men 66 to 96 years old (mean age 79 years) who were segregated independently as a detrusor overactivity group by prospective urodynamic evaluation. All but 1 patient had incontinence and/or other symptoms, and none had diabetes or a significant neurological deficit. The dysjunction pattern was characterized by moderately widened intercellular spaces, scarce intermediate muscle cell junctions, abundant distinctive protrusion junctions and ultra-close cell abutments, and absence of profiles characteristic of enlarged hypertrophic cells. There was superimposed widespread degeneration of muscle cells and axons in 8 specimens, which matched the subgroup of patients with impaired detrusor contractility. The remaining 7 specimens with no degeneration matched the patients with normal contractility. Protrusion junctions and abutments are proposed as a possible manifestation of a process of muscle cell de differentiation associated with natural aging, as well as the mediator in overactive detrusor of electrical coupling of muscle cells, in lieu of their normal mechanical coupling curtailed by marked reduction of intermediate cell junctions. On this basis, a bipartite myogenic mechanism is proposed to account for the involuntary contractions yet allow neurally triggered unitary voiding contractions in the overactive detrusor. Superimposed degeneration is proposed as the structural basis of impaired detrusor contractility, when also present. PMID- 8411456 TI - Structural basis of geriatric voiding dysfunction. IV. Bladder outlet obstruction. AB - Several aspects of the pathogenesis of voiding dysfunction in bladder outlet obstruction remain unresolved. The structural basis of obstructive versus nonobstructive dysfunction was investigated in a prospective ultrastructural/urodynamic study of 35 elderly subjects of comparable age. Detrusor structure was examined by electron microscopy, with blinded clinical and urodynamic information. Seven detrusor specimens were segregated by a distinctive myohypertrophy, structural pattern, which matched with 6 male and 1 female subjects 72 to 96 years old (mean age 83) who had urodynamically proved outlet obstruction. This pattern was characterized by widely separated muscle cells with reduction of intermediate cell junctions, collagenosis, that is abundant collagen plus some elastic fibers, in the markedly widened spaces between individual muscle cells and abundant profiles characteristic of enlarged, hypertrophic muscle cells. Superimposed degeneration of muscle cells and axons in 6 specimens matched those of 5 men and 1 woman who had impaired detrusor contractility. In 3 specimens there were also abundant protrusion junctions and ultra-close abutments; these matched those of 2 men and 1 woman with obstruction plus detrusor overactivity. Observations on the degree of bladder trabeculation in the entire population of 35 subjects are presented. It is concluded that bladder outlet obstruction is associated with changes in detrusor structure that can account for the resultant voiding dysfunction. Features of the myohypertrophy pattern, with or without superimposed degeneration, can explain overall weakness of the obstructed detrusor despite hypertrophy of its cells. Protrusion junctions and abutments probably mediate electrical coupling of muscle cells leading to involuntary contractions in the overactive (unstable) obstructed detrusor. Excessive deposits of elastic fibers (hyperelastosis) between widely separated muscle cells and in interstitium are suggested as the probable structural basis for increased bladder distensibility and chronic retention. PMID- 8411457 TI - Penetration of ciprofloxacin into prostatic fluid, ejaculate and seminal fluid in volunteers after an oral dose of 750 mg. AB - To evaluate an effective dose for the treatment of bacterial prostatitis the concentrations of ciprofloxacin were measured in prostatic fluid, ejaculate and the cell-free seminal fluid of 15 healthy volunteers who received an oral dose of 750 mg. ciprofloxacin while in a fasting state. Venous blood samples were taken in all subjects at 1, 2, 3 and 4 hours. In 6 subjects blood samples were also taken after 8 and 12 hours. Urine was collected in all subjects during 0 to 4 hours and in the 6 subjects also during 4 to 8 hours and 8 to 12 hours. Prostatic fluid could be obtained in 10 subjects by prostatic massage 4 hours after drug intake. So as not to contaminate the urethra with ciprofloxacin the subjects were not allowed to void until 4 hours after drug intake. Iopamidol (3.162 gm.), a renal contrast agent, was administered intravenously concomitantly with oral ciprofloxacin intake. After 8 hours iohexol (3.235 gm.) was administered intravenously. These agents were measured in prostatic fluid, ejaculate and seminal fluid to assess the contamination of those fluids by urine. All drug measurements were done by high pressure liquid chromatography. The median plasma concentrations of ciprofloxacin were 2.1 mg./l. at 1 hour (maximum concentration), 0.9 mg./l. at 4 hours and 0.2 mg./l. at 12 hours. The median concentration in prostatic fluid was 0.23 mg./l. with a fluid-to-plasma concentration ratio of 0.23. The median concentration in the ejaculate (seminal fluid) after 4 hours was 7.4 mg./l. (6.6 mg./l.) and after 12 hours it was 2.0 mg./l. (1.9 mg./l.) with corresponding ejaculate (seminal fluid)-to-plasma concentration ratios of 8.4 (7.7) and 8.0 (6.6), respectively. Thus, ciprofloxacin is concentrated several-fold in ejaculate and seminal fluid but not in prostatic fluid. According to the results the concentrations of ciprofloxacin in prostatic fluid exceed the minimal inhibitory concentration-90% for Enterobacteriaceae but not for Pseudomonas, enterococci and staphylococci, whereas the concentrations in ejaculate and seminal fluid are sufficiently elevated to include the total spectrum of sensitive strains causing bacterial prostatis. PMID- 8411458 TI - Norfloxacin as prophylaxis against urethral strictures following transurethral resection of the prostate: an open, prospective, randomized study. AB - An open, prospective, randomized study was performed to investigate the effect of norfloxacin prophylaxis on stricture formation and operative outcome after transurethral resection of the prostate. After resection, the 359 patients studied were randomly divided into 2 groups: 1) those given norfloxacin as prophylaxis for 15 days following removal of the catheter (norfloxacin group) and 2) those given no antimicrobial prophylaxis during the same period (control group). Of the patients 94 were excluded. At followup 6 to 12 months postoperatively, the number of strictures in the anterior urethra was 2 of 135 in the norfloxacin group and 22 of 130 in the control group (p < 0.01). Strictures in the bladder neck developed in 3 of 135 and 4 of 130 patients, respectively (not significant). As a consequence of a lower structure incidence in the anterior urethra in the norfloxacin group, fewer patients in that group were dissatisfied with the operative outcome. The results suggest that norfloxacin provides effective prophylaxis against stricture formation after transurethral resection of the prostate. PMID- 8411459 TI - Newly diagnosed bulbar urethral strictures: etiology and outcome of various treatments. AB - A retrospective review was performed of 199 consecutive patients who were evaluated at this institution between 1976 and 1990 because of a newly diagnosed bulbar urethral stricture. Mean patient age at diagnosis was 64 years (range 10 to 96) and most patients presented with obstructive symptoms. The stricture etiology was primarily iatrogenic (47%), secondary to a transurethral procedure. The strictures were usually short (less than 2 cm., 96%), single (99%) and located in the proximal bulb (57%). Of the 151 patients receiving treatment at the time of initial diagnosis 101 (67%) underwent urethral dilation, 39 (26%) were managed with direct vision internal urethrotomy and in 11 (7%) a cystotomy tube was placed. With a median followup of 3.5 years (range 0 to 16), there was an estimated retreatment rate of 2.4 treatments per 10 person-years. The probability of not requiring retreatment within 3 years was 65 +/- 5% for urethral dilation and 68 +/- 8% for direct vision internal urethrotomy. When compared to urethral dilation, direct vision internal urethrotomy resulted in a higher incidence of postprocedure cystitis (5% versus 3%), epididymitis (5% versus 3%) and penile hemorrhage (8% versus 2%). These findings indicate that both conservative therapies were equally efficacious as an initial treatment of bulbar urethral stricture. However, direct vision internal urethrotomy did have a slightly higher complication rate. No specific patient or stricture characteristics could be identified that were reliable for predicting therapeutic outcome. PMID- 8411460 TI - Treatment of sphincter strictures using permanent UroLume stent. AB - The success of the permanent UroLume urethral stent in the treatment of recurrent bulbar urethral strictures led to its use in cases of recurrent strictures in the sphincter active membranous urethra. Among 6 patients treated 3 traumatic strictures have recurred despite the urethral stent but 3 post-prostatectomy strictures have been treated successfully in conjunction with an artificial sphincter to restore continence. PMID- 8411461 TI - Prophylactic antibiotics in transurethral surgery. PMID- 8411462 TI - Allograft vein bypass: is it an acceptable alternative for infrapopliteal revascularization? AB - PURPOSE: Autogenous vein bypass grafts to infrapopliteal outflow sites have patency and limb salvage rates significantly superior to those obtained with prosthetic grafts. However, when infrageniculate bypass is required for limb threatening ischemia in the patient lacking suitable autogenous veins, nonautogenous reconstruction or primary amputation are the only other alternatives. METHODS: During a 2-year period we implanted 25 cryopreserved allograft saphenous vein bypass grafts in 24 patients (median age 76 years) with tissue necrosis (20 patients), rest pain (4 patients), or acute ischemia (1 patient); 16 patients were men and 8 were women. As many as six previous revascularizations were performed in 79%; two grafts extended to the infrageniculate popliteal artery; 23 grafts extended to a paramalleolar vessel. RESULTS: Secondary patency at 1 month was 87%, but only 36% at 1 year. Use of warfarin (Coumadin) failed to improve the patency rate (five of nine occlusions treated with Coumadin versus eight of 16 not treated with Coumadin). Only eight of 24 patients are alive with open grafts; nine patients have died. CONCLUSIONS: Unheralded occlusions more typical of prosthetic graft failure tempered the initial enthusiasm and effectiveness of vein allografts. All autogenous options must be exhausted to complete distal, secondary revascularization before resorting to nonautogenous conduits. Use of allograft veins must be viewed with continued skepticism. PMID- 8411463 TI - Early experience with cryopreserved saphenous vein allografts as a conduit for complex limb-salvage procedures. AB - PURPOSE: The lack of a suitable alternative to autogenous vein is often the limiting factor for complex lower extremity vascular reconstruction, especially when previously placed grafts have failed. Cryopreserved saphenous vein allografts have been used as an alternative conduit. This report reviews our early experience with this conduit in a series of complex redo revascularization procedures for limb salvage when no suitable autogenous vein was available. METHODS: Thirty-five patients underwent 39 lower extremity bypass grafts on 36 limbs. These patients had undergone a combined total of 72 prior revascularization procedures on the symptomatic limb, an average of two procedures per patient. Only five bypasses were performed as a primary procedure. There were 18 men and 17 women with a mean age of 71 years. Sixteen of the patients had diabetes. Thirty-four bypasses were performed for rest pain or ulceration, four for disabling claudication, and one for replacement of an aneurysmal vein graft. There were 35 femorotibial, three below-knee femoropopliteal, and one femoropedal reconstruction. Twenty-five grafts were constructed with cryopreserved vein only, whereas 14 were composite grafts; 10 were constructed with polytetrafluoroethylene, one with Dacron, and three with spliced native saphenous vein. The mean follow-up was 9 months (range 1 to 25 months). RESULTS: There was one early death (< 30 days) and two late deaths. Two patients died with a patent graft. There have been 12 early graft closures and an additional 17 late failures, resulting in primary cumulative graft patency rates of 67%, 56%, 43%, 28%, and 14% at 1, 3, 6, 12, and 18 months, respectively. Surgically correctable causes, including technical error and anastomotic stenosis, could be identified in 13 of the 29 graft failures. Salvage of failed grafts resulted in secondary cumulative graft patency rates of 87%, 77%, 61%, 46%, and 37% at these same intervals. There was no significant difference in primary or secondary graft patency rates related to diabetes, ABO graft compatibility, graft composition or orientation, indication for surgery, state of the outflow tract, or site of distal anastomosis. Limb salvage was attained in 24 (67%) of the 36 limbs. Two amputations were necessary despite patent grafts. CONCLUSIONS: Because of the poor overall graft patency rates, cryopreserved saphenous vein allografts should be used only as a last resort when no alternative autogenous conduit is available. Unless patency rates superior to those achievable with currently available prosthetic or biologic conduits can be attained by adjunctive measures such as routine anticoagulation or immunosuppressive therapy, the use of cryopreserved saphenous vein allografts for lower extremity revascularization should be deferred until improved preparation techniques provide a more durable conduit. PMID- 8411464 TI - Failure of glow-discharge polymerization onto woven Dacron to improve performance of hemodialysis grafts. AB - PURPOSE: The ideal conduit for hemodialysis vascular access remains elusive. Autogenous fistulas and prosthetic grafts, most commonly expanded polytetrafluoroethylene (e-PTFE), have adequate long-term patency rates (60% to 80% at 1 year); however, considerable delay in their use (2 to 6 weeks) is required. The Plasma-TFE graft is a recently introduced thin-walled woven Dacron graft to which an ultrathin layer of tetrafluoroethylene is bonded through a process of glow-discharge polymerization. This process purportedly results in a graft with an internal surface of low thrombogenicity. Low thrombogenicity, combined with the healing characteristics of a woven graft, have led to claims of equivalent patency rates even when used for dialysis immediately (within 1 week) after implantation. METHODS: This concept led us to use this new graft material in 19 fistulas (12 forearm and 7 arm) during a 1-year period. RESULTS: Although early use was possible, the primary and secondary patency rate at 12 months was only 47.4%. Ten grafts required replacement, five within the first month and two in the second month. Attempts at fistula revision failed because of unsuccessful graft thrombectomy or exuberant intimal hyperplasia. Failure was not associated with early use. During the same time period, 28 PTFE grafts were implanted, with only four failures (primary patency 78.6%; secondary patency 85.7%; p = 0.028). The secondary patency rate was the same for Plasma-TFE grafts (47%) but improved to 85.7% for e-PTFE grafts (p = 0.005). Both groups were comparable with respect to age, diabetes, previous dialysis access procedures, and other comorbid conditions. CONCLUSIONS: These early results have been sufficiently disappointing that we have abandoned use of this graft approved for hemodialysis by the Food and Drug Administration and cannot recommend it for other clinical indications. Nevertheless, the concept of plasma-discharge polymerization is theoretically attractive and might be useful in future graft configurations. PMID- 8411465 TI - Hemodynamic evaluation of a bioprosthetic venous prosthesis. AB - PURPOSE: A prosthetic venous valve must be biocompatible and nonthrombogenic and function in the venous circulation. Biocompatibility and thrombogenicity of our prosthesis have been examined in prior animal experiments, and 91% of valve conduits including early prototypes are patent at 3 weeks. However, evaluation of valve function is much more difficult in animals; therefore in this study the function of excised valves was evaluated ex vivo. METHODS: Nine bovine jugular vein conduits, each with one bileaflet venous valve, were harvested and placed in a venous flow simulator. Flows and pressures were adjusted to mimic human respiratory and hydrostatic variations. Each valve and conduit was tested for variations in valve diameter and sinus expansion in response to flow. Valve opening and closing times and valve competence were measured in response to pressure changes. After testing, each specimen was glutaraldehyde fixed and assessed a second time. RESULTS: Valve orifice area increased in response to flow in both fresh and fixed tissues. Maximum valve orifice area was reduced by fixation (27.7%) at full flows (p < 0.05). Valve sinus dimensions increased in response to increased pressure until maximum expansion was achieved (33 mm Hg). This was reduced 15.3% in fixed tissue (p < 0.05). Valve opening times (at < 1 mm Hg gradient) were slightly longer in fixed compared with fresh tissue (0.43 +/- 0.09 vs 0.41 +/- 0.13 second; p < 0.05). Valve closing times were comparable in both states (0.43 +/- 0.08 vs 0.49 +/- 0.07 second). Three fresh and seven fixed specimens that were subjected to 287 mm Hg back pressure exhibited minimal reflux. CONCLUSIONS: Size and availability make the bovine jugular vein valve an ideal venous valve substitute. Glutaraldehyde fixation renders the tissue biocompatible and nonthrombogenic while preserving anatomic integrity and leaflet strength and flexibility. Mounted and stented in a sewing sleeve, this prosthesis could represent the first generally applicable clinical solution to chronic venous insufficiency and venous hypertension. PMID- 8411466 TI - Platelet aggregometry can accurately predict failure of externally supported knitted Dacron femoropopliteal bypass grafts. AB - PURPOSE: Our purpose was to evaluate whether a method for quantification of platelet aggregability will predict failure of knitted Dacron femoropopliteal bypass grafts. METHODS: A numerically derived platelet aggregation (PA) score, based on the aggregation pattern and platelet count, was determined in the 40 patients available for platelet analysis who underwent 53 femoropopliteal bypass grafts with preclotted, 6 mm, externally supported knitted Dacron grafts from 1981 to 1991 (mean follow-up 50 months). The preoperative score was found to remain stable after surgery, enabling the use of postoperative values when preoperative values were not available. The PA score was available in 19 patients (23 grafts) before surgery and 23 patients (30 grafts) after surgery. The following factors were analyzed for predicting graft failure by the Cox proportional hazards regression model: PA score, age, gender, history of smoking, coronary artery disease, hypertension, hyperlipidemia, cerebrovascular disease, diabetes, claudication versus limb salvage, site of the distal anastomosis, previous ipsilateral bypass, and state of the runoff as determined by preoperative angiography. RESULTS: Of the studied risk factors, the value of the PA score was the most significant predictor of graft closure (p < 0.0001). An increase of 10 units was associated with an increased relative risk of 2.02. Throughout the follow-up period, 15 of 16 grafts remained patent in patients with a PA score of 15 or less, but only 2 grafts out of 17 remained patent in patients with a PA score of 30 or greater. CONCLUSIONS: These data suggest that the PA score is a potential risk factor for failure of femoropopliteal bypass with externally supported knitted Dacron grafts. PMID- 8411467 TI - Deep venous insufficiency: the relationship between lysis and subsequent reflux. AB - PURPOSE: Although venous valvular insufficiency is well recognized as the most important etiologic mechanism in the development of the postthrombotic syndrome, the factors contributing to valve incompetence after deep venous thrombosis remain obscure. METHODS: To establish the relationship between recanalization and valve competence, 113 patients with acute deep venous thrombosis were studied with serial duplex ultrasonography. RESULTS: Median lysis times for segments developing reflux (214 to 474 days) were 2.3 to 7.3 times longer than for corresponding segments not developing reflux (65 to 130 days) for all except the posterior tibial vein. In the posterior tibial vein, median lysis times for those with and without reflux were nearly identical (72 vs 80 days). The median time to onset of reflux was significantly less than the median lysis time in the mid and distal superficial femoral veins and was simultaneous with recanalization in all other segments. CONCLUSIONS: Early recanalization is important in preserving valve integrity for all but the posterior tibial segment. However, the small number of patients with reflux despite early lysis (< 1 month) or without reflux despite relatively late lysis (> 9 to 12 months) suggests that other factors may also contribute to the development of valvular incompetence. These factors may be particularly important in the posterior tibial vein, in which lysis time has little relationship to the ultimate development of reflux. PMID- 8411468 TI - Comparison of cardiac morbidity rates between aortic and infrainguinal operations: two-year follow-up. Study of Perioperative Ischemia Research Group. AB - PURPOSE: We have previously prospectively compared the differences in perioperative cardiac ischemic events in 140 patients undergoing major abdominal (n = 53) versus infrainguinal (n = 87) vascular operations. This study was designed to extend these observations by determining the 2-year cardiac prognosis of patients at high risk undergoing abdominal aortic versus infrainguinal vascular operations. METHODS: Data included historical, clinical, and laboratory data collected during the in-hospital period, and at 6 months, 1 year, and 2 years after surgery. This information was collected independently of the usual clinical care visits. Data were analyzed with Cox's proportional hazards model. RESULTS: There were 11 in-hospital deaths overall (five [9%] aortic; six [7%]) infrainguinal). 628 days (median 726 days). Fifteen patients (12%) had fatal myocardial infarctions, two (4%) of which occurred in patients who underwent aortic procedures and 13 (16%) of which occurred in patients who underwent infrainguinal operations. Nonfatal myocardial infarctions befell one (2%) patient undergoing aortic surgery and four (5%) patients undergoing infrainguinal surgery. One (2%) patient undergoing aortic surgery and three (4%) patients undergoing infrainguinal surgery were admitted to the hospital with unstable angina during the follow-up period. In all, adverse cardiac outcomes occurred in 20 of 81 (25%) patients who had infrainguinal procedures compared with four of 48 (8%) patients who had aortic operations (p = 0.04). Multivariate analysis showed that a history of diabetes (p = 0.001) and definite coronary artery disease (p = 0.01) are independently associated with adverse outcomes after both types of peripheral vascular operations. CONCLUSIONS: The incidence of long-term adverse cardiac outcomes in patients at high risk undergoing infrainguinal operations is substantially greater than in those undergoing aortic operations, mostly because of a greater prevalence of diabetes, and definite coronary artery disease in the former group. PMID- 8411469 TI - Surgery for chronic lower extremity ischemia in patients eighty or more years of age: operative results and assessment of postoperative independence. AB - PURPOSE: Revascularization for chronic lower extremity ischemia (CLEI) in patients 80 or more years of age is controversial. High operative mortality, short remaining life span, and nonambulatory nursing home existence are cited as frequent outcomes in this patient group. Despite these considerations we have maintained an aggressive policy of lower extremity revascularization in functional and potentially functional patients 80 or more years of age with limb threatening ischemia or claudication sufficient to threaten their independence. METHODS: The records of all patients 80 or more years of age who underwent lower extremity revascularization for CLEI from 1981 through 1991 were reviewed. Patients were evaluated for surgical indication, perioperative mortality, late survival, graft patency, and limb salvage. In addition, the patient's preoperative and postoperative independence determined by living situation (home versus nursing home) and ambulatory status were examined. RESULTS: During the period reviewed 88 patients 80 or more years of age underwent 95 arterial reconstructions for CLEI; 95% for limb salvage and 5% for short-distance claudication. There were 81 (85%) infrainguinal bypasses and 14 (15%) extra anatomic bypasses, 5 of which also included a simultaneously placed infrainguinal bypass. The perioperative mortality rate was 6%. Mean hospitalization was 18 days (range 6 to 62). The 1- and 3-year primary graft patency rates were 88% and 66%, limb salvage rates 94% and 91%, and patient survival rates 73% and 51%, respectively. One year after operation 88% of patients were ambulatory, 85% were living at home, and 80% were both living at home and ambulatory. At 3 years these results were 86%, 76%, and 71%, respectively. Of the patients living at home and ambulatory before operation, 100% and 84% of survivors were still living at home and ambulatory 1 and 3 years after operation. Of patients living at home who had late graft occlusions, 67% required amputation and subsequent nursing home placement. CONCLUSION: Although survival in this group is predictably lower than that of age-matched control subjects, octogenarians have satisfactory 1- and 3 year graft patency, limb salvage, and functional results. Revascularization for CLEI in patients 80 or more years of age is appropriate and results in maintenance of independent living in a large majority. PMID- 8411470 TI - Outcome of infrainguinal arterial reconstruction in women. AB - PURPOSE: The outcome of leg bypass in women is unknown. To date, most series of lower extremity bypass have included few women, and the results in women compared with those in men have not been reported. The experience with infrainguinal arterial reconstruction in women treated at the Oregon Health Sciences University has been greater than previously reported. We have reviewed our recent experience with lower extremity bypass to determine whether results in women differ significantly from those in men. METHODS: In the past 11 years we have performed 823 infrainguinal arterial reconstructions for lower extremity ischemia in 585 patients, of which 357 procedures were performed in women and 466 procedures were performed in men. The mean ages were 65 years for men and 68 years for women. Diabetes was present in 59% of the men and 48% of the women. Among men, 84% had a history of tobacco use compared with 66% of the women. Bypasses in men were performed for limb salvage in 73%, claudication in 22%, and a failing bypass graft in 5% of cases. Bypasses in women were performed for limb salvage in 79%, claudication in 15%, and a failing graft in 6% of cases. Previous revascularizations had been performed in 63% of the men and 71% of the women. Autogenous vein was used in 97% of the bypasses in men and 96% of bypasses in women. Graft distribution among infrainguinal arterial target sites was similar between the groups. Bypasses in men were femoral to above-knee popliteal in 11%, femoral to below-knee popliteal in 40%, femoral to tibial in 38%, popliteal to tibial in 10%, and tibial to tibial in 1% of cases. Bypasses in women were femoral to above-knee popliteal in 14%, femoral to below-knee popliteal in 43%, femoral to tibial in 33%, popliteal to tibial in 9%, and tibial to tibial in 1% of cases. RESULTS: Perioperative 30-day mortality rates were 3.7% overall, 4.3% in men, and 2.9% in women. Long-term survival at 1, 3, and 5 years in men was 80%, 59%, and 44%, respectively. Long-term survival at 1, 3, and 5 years in women was 83%, 69%, and 44%, respectively. Life-table primary patency rates at 1, 3, and 5 years were 86%, 77%, and 71% for men and 87%, 74%, and 67% for women. Limb salvage results at 1 and 5 years were 93% and 91% for men and 96% and 96% for women. CONCLUSIONS: These results indicate that long-term graft patency and limb salvage results in women are identical to those obtained in men in this experience with autogenous vein. Infrainguinal arterial reconstruction can be performed in women with mortality rates similar to those of men. In this series long-term survival was similar for both men and women. PMID- 8411471 TI - Tourniquet occlusion technique for tibial artery reconstruction. AB - PURPOSE: Vascular clamps, vessel loops, and intraluminal occluding devices used to control tibial and pedal vessels can be injurious and may fail to occlude heavily calcified arteries. In an effort to prevent injury to these small arteries, we have investigated the safety and efficacy of thigh pneumatic tourniquet occlusion for distal vascular control during infrapopliteal reconstruction. METHODS: During an 18-month period, 88 tibial or pedal arterial reconstructions were performed on 80 patients (mean age 75 years) by the tourniquet technique. Data regarding indications for operation, preoperative evaluation, intraoperative findings, surgical technique, and early outcome were recorded prospectively. RESULTS: Sixty percent of patients were diabetic: 36% insulin dependent and 24% non-insulin dependent. The indications for operation were claudication in 6 (7%), ischemic ulcer in 24 (27%), rest pain in 25 (28%), and gangrene in 33 (38%) patients. Thirty-five percent of operations followed failed ipsilateral infrainguinal reconstructions. The peroneal artery was the target vessel in 38%, anterior tibial in 26%, posterior tibial in 23%, tibioperoneal trunk in 9%, and inframalleolar vessels in 4% of cases. Preoperative analog waveforms and ankle-brachial indexes were used to classify the tibial arteries as compliant, 49%; relatively noncompressible, 30%; and absolutely noncompressible, 9%. Twelve percent had no Doppler flow at the ankle level. At operation 36 of the target arteries (41%) had mural calcification. Tourniquet pressures of 200 to 400 mm Hg (mode 250 mm Hg) were applied from 13 to 55 minutes (mean 27.1 +/- 9.1 minutes). All patients were given systemic anticoagulants. In 19 limbs (22%) the tourniquet was used to occlude a patent superficial femoral artery above the proximal (inflow) anastomosis to either the superficial femoral artery (8%), the above-knee popliteal artery (5%), or the below-knee popliteal artery (9%). Hemostasis was adequate in all cases and no alternative occlusive devices were required. There were no significant complications attributable to the use of the pneumatic tourniquet. CONCLUSION: Tourniquet occlusion simplifies the infrapopliteal dissection, lessens operating time, improves visualization of the distal anastomosis, and removes the potential for arterial injury to the target vessel. Arterial calcification and noncompressible tibial arteries do not contraindicate the use of thigh tourniquet occlusion. This technique is preferred for all patients undergoing tibial or pedal artery reconstruction. PMID- 8411472 TI - Limb salvage and wound coverage in patients with large ischemic ulcers: a multidisciplinary approach with revascularization and free tissue transfer. AB - PURPOSE: Large ischemic wounds, particularly with exposed bone or tendons, may not heal even after successful revascularization. We have taken an aggressive approach for limb salvage that uses autogenous vein grafting and simultaneous microvascular free tissue transfer. METHODS: In the past year, seven patients (average age 67 years; range 56 to 79) with ischemic disease and distal ulceration underwent revascularization for limb salvage and free tissue transfer. Each had a nonhealing wound (average size 80 cm2), present for 8.6 months (range 2 to 24 months). Simultaneous vein bypass and free tissue transfer was performed in four (57%) of the seven patients. RESULTS: All flaps were initially viable; however, one was lost on day 4 because of hypotension and congestive heart failure. One patient with a successful flap died at 1 month of pneumonia. Minor wound complications were seen in four (57%) of seven patients. Five of the seven patients had the wounds heal completely and are ambulatory at an average follow up of 10 months. CONCLUSIONS: Our aggressive approach was successful in preserving limb length and function in 71% of our patients. We perform simultaneous procedures whenever possible to minimize operative and hospitalization times. We believe that this combined approach optimizes the treatment of ischemic limbs with large ulcers. PMID- 8411473 TI - Three-dimensional spiral computed tomographic angiography: an alternative imaging modality for the abdominal aorta and its branches. AB - PURPOSE: We sought to apply a new technique of computed tomographic angiography (CTA) to the preoperative and postoperative assessment of the abdominal aorta and its branches. METHODS: After a peripheral intravenous contrast injection, the patient is continuously advanced through a spiral CT scanner, while maintaining a 30-second breath-hold. Thirty-five patients with abdominal aortic, renal, and visceral arterial disease have undergone CTA. RESULTS: Diagnostic three dimensional images were obtained in patients with aortic aneurysms (n = 9), aortic dissections (n = 4), and mesenteric artery stenoses (n = 4). The technique has also been used to assess vessels after operative reconstruction or endovascular intervention in 18 patients. These preliminary studies have correlated well with conventional arteriographic findings. In aneurysmal disease both the lumen and mural thrombus and associated renal artery stenoses are visualized. The true and false channels of aortic dissections and the perfusion source of the visceral vessels are clearly shown; patency of visceral and renal reconstruction or stent placement are confirmed. CTA is relatively noninvasive and can be completed in less time than conventional angiography with less radiation exposure. CONCLUSIONS: This initial experience suggests that CTA may be a valuable alternative to conventional arteriography in the evaluation of the aorta and its branches. PMID- 8411474 TI - Benefit of carotid endarterectomy after prior stroke. AB - PURPOSE: The benefit of carotid endarterectomy (CEA) in preventing recurrent stroke in patients who have sustained a previous stroke remains controversial. The purpose of this study was to evaluate the immediate results and long-term benefit of CEA after recovery from a prior ipsilateral stroke. METHODS: Between 1980 and 1990, 85 patients underwent CEA for prior stroke with an average follow up of 54 months (range 0.3 to 130). The interval from prior stroke to CEA averaged 19.8 months (range 0.1 to 158.3). Vascular risk factors included hypertension in 70.6%, diabetes in 20.0%, history of smoking in 80.0%, and associated coronary artery disease in 51.8% of the patients. RESULTS: There were no perioperative deaths. Four patients (4.7%) had an ipsilateral stroke within 30 days of operation. During the follow up 31 patients (36.5%) died. The leading cause of death was cardiac (54.8%) followed by cancer (16.1%). By life-table methods, the cumulative incidence of recurrent stroke at 9 years was 14% for strokes in the ipsilateral distribution, for an annual stroke risk of 1.6% per year. The cumulative incidence of stroke in other distributions was 5%, for an annual stroke risk of 2.1% per year for all strokes. When interval to operation, preoperative stroke severity, vascular risk factors, and neurologic symptoms were evaluated, no independent indicator of increased risk of recurrent postoperative stroke could be identified. CONCLUSION: These results demonstrate a marked improvement over the natural history and best medical therapy for these lesions as reoperated in the literature. We conclude that CEA is beneficial in preventing recurrent stroke in this group of patients and should be considered the appropriate management in this setting. PMID- 8411475 TI - The role of duplex scanning and arteriography before carotid endarterectomy: a prospective study. AB - PURPOSE: This study examines the current role of diagnostic tests done before carotid endarterectomy and the need for routine arteriography. METHODS: We prospectively studied vascular surgeons' decision-making over a 29-month period during which 111 carotid arteries in 103 patients were considered for endarterectomy. For each case the surgeon's management plan was recorded after clinical evaluation and review of the duplex scan findings, but before arteriography. This plan was later compared with the patient's ultimate clinical management. RESULTS: Of 111 total cases in this period, 17 were excluded from analysis because arteriography was not done or it was performed before the surgeon's evaluation. Carotid duplex scans were diagnostic in 87 (93%) of the remaining 94 cases. The carotid lesion was incompletely assessed by duplex scanning in seven patients because the disease was not limited to the distal common or proximal internal carotid artery (n = 4); anatomic or pathologic features of the carotid artery interfered with imaging or accurate Doppler assessment (n = 1); or an internal carotid artery occlusion could not be distinguished from a high-grade stenosis (n = 2). When a technically adequate duplex scan showed significant disease of the carotid bifurcation, arteriography contributed information that affected clinical management in only a single case (1%). This patient had a middle cerebral artery occlusion distal to a high-grade carotid bifurcation stenosis. CONCLUSIONS: Clinical assessment and duplex scanning were sufficient for the preoperative evaluation of 93% of the candidates for carotid endarterectomy. Clinical circumstances or atypical duplex scan findings can be used to identify the minority of patients for whom arteriography is necessary. On the basis of this experience, we have developed practical guidelines for the selective use of arteriography before carotid endarterectomy. PMID- 8411476 TI - Early and late outcome of surgical repair for small abdominal aortic aneurysms: a population-based analysis. AB - PURPOSE: Whether small abdominal aortic aneurysms (AAAs) (< or = 5 cm in diameter) should be repaired early to enhance late survival remains controversial. Long-term population-based data on the surgical outcome for small AAAs may help to establish practice guidelines until randomized clinical trials are completed. METHODS: To examine an entire community experience with small AAAs, we conducted a population-based analysis of the recognition, reasons for operation, perioperative mortality rates, and late survival in Olmsted County, Minnesota. RESULTS: The incidence of recognized small AAAs increased thirtyfold during a 30-year period. The propensity to repair small AAAs also increased during the same period. Eventually one third of small AAAs were repaired. The primary reasons for surgical consultation and operation were presence of the aneurysm (49%), expansion on serial examination (28%), nonspecific abdominal or back symptoms (18%), and excessive patient anxiety about the aneurysm (5%). Community operative mortality rates for small AAAs were low (2.6%) compared with those for large aneurysms (5.5%) (p = 0.65). However, the observed 5-year survival rate for the group undergoing repair of small aneurysms was 62%, which was significantly less than the 83% expected survival for the general population (p < 0.05). Relative survival for the operated small and large aneurysms was similar. The primary cause of death for both groups was myocardial infarction. CONCLUSIONS: The results of our population-based analysis indicate that early operative results for elective repair of small AAAs are excellent, but late survival remains significantly impaired by coronary heart disease. Consequently, our data question whether early repair of small AAAs will enhance late survival. Until randomized clinical trials on management of small AAAs are completed, most small AAAs should be monitored for expansion and operated on electively when they approach or enter the range of 5 to 6 cm in good-risk patients. PMID- 8411477 TI - The resource-based relative value scale in vascular surgery. A report of the activities of the Joint Council of the Society for Vascular Surgery and the North American Chapter of the International Society for Cardiovascular Surgery in Medicare reimbursement reform. PMID- 8411478 TI - Costs of duplex scanning and the impact of the changes in Medicare reimbursement. AB - PURPOSE: To obtain objective cost data to use in a formal comment on the proposed changes in Medicare reimbursement, a mail survey of vascular laboratories was carried out. METHODS: Data were received from 142 facilities. Patients receiving Medicare made up 64% of the volume, and duplex scanning accounted for 78% of the work. RESULTS: The mean cost per scan, exclusive of physician payments, was $181. When compared with the mean allowable reimbursement under the 1992 Fee Schedule of $113, this represents a loss of $68 for each duplex scan performed on a patient receiving Medicare. In general, there was little difference in costs when the data were analyzed on the basis of type or size of facility. For laboratories providing data on 1991 Medicare reimbursements, the new fee schedule results in a 38% drop in payments. CONCLUSIONS: These data substantiate the impression that the Health Care Finance Administration has substantially undervalued the costs of performing duplex scanning. PMID- 8411479 TI - Popliteal venous aneurysm: report of two cases and review of the world literature. AB - Two new cases of popliteal venous aneurysm are reported and added to the 22 other cases of popliteal venous aneurysm available for review. Both patients were first seen with acute pulmonary embolism and were treated with thrombolytic therapy followed by anticoagulation. Each had recurrent venous thromboembolism before discovery of the popliteal venous aneurysm. One popliteal venous aneurysm was diagnosed with phlebography and the second with venous duplex imaging, confirmed with phlebography. Both were surgically corrected with tangential aneurysmectomy and lateral venorrhaphy. Twenty-four cases of popliteal venous aneurysm are now available for review. Seventy-one percent (17 of 24) presented with pulmonary embolism, 88% (21 of 24) were saccular, and 96% (23 of 24) were located in the proximal popliteal vein. All but two were diagnosed by ascending phlebography. Three patients received no treatment: in two of these the outcome was not documented and the third had occasional pain. Two patients received anticoagulation without subsequent operative repair and both died of recurrent pulmonary emboli. Operative correction resulted in a 75% patency rate with 21% complications, most of which were related to postoperative anticoagulation. No patient who was operated on had subsequent pulmonary embolism, and there were no operative deaths. We suggest that all patients who have pulmonary embolism have lower-extremity venous duplex imaging. All popliteal venous aneurysms should be surgically repaired, inasmuch as nonoperative therapy results in recurrent thromboembolism and an unacceptably high mortality rate. Tangential aneurysmectomy with lateral venorrhaphy is the recommended procedure. PMID- 8411480 TI - Regarding "Venous ulcers and the superficial venous system". PMID- 8411481 TI - A piece of my mind. Which hat do I wear? PMID- 8411482 TI - Finding neural tube 'zippers' may let geneticists tailor prevention of defects. PMID- 8411483 TI - Women need more and better information on menopause from their physicians, says survey. PMID- 8411484 TI - Medicare payment dispute may stymie flu shots. PMID- 8411485 TI - From the Food and Drug Administration. PMID- 8411486 TI - From the Institute of Medicine. PMID- 8411487 TI - From the Centers for Disease Control and Prevention. Evaluation of surveillance for Chlamydia trachomatis infections in the United States, 1987 to 1991. PMID- 8411488 TI - From the Centers for Disease Control and Prevention. Handwashing and glove use in a long-term-care facility--Maryland, 1992. PMID- 8411489 TI - Active and passive smoking and pathological indicators of lung cancer--a report of limited value? PMID- 8411490 TI - Active and passive smoking and pathological indicators of lung cancer--a report of limited value? PMID- 8411491 TI - Active and passive smoking and pathological indicators of lung cancer--a report of limited value? PMID- 8411492 TI - Active and passive smoking and pathological indicators of lung cancer--a report of limited value? PMID- 8411493 TI - Active and passive smoking and pathological indicators of lung cancer--a report of limited value? PMID- 8411494 TI - Treatment strategies for prostate cancer. PMID- 8411495 TI - Treatment strategies for prostate cancer. PMID- 8411496 TI - Treatment strategies for prostate cancer. PMID- 8411497 TI - Treatment strategies for prostate cancer. PMID- 8411498 TI - Growth hormone therapy for the elderly: the fountain of youth proves toxic. PMID- 8411499 TI - Do the poor sue more? A case-control study of malpractice claims and socioeconomic status. AB - OBJECTIVE: To evaluate whether socioeconomic status is associated with risk of malpractice claims, particularly among those who have suffered medical injury. DESIGN: Case-control study. SETTING: Fifty-one hospitals in New York State. METHODS: The presence and severity of medical injury, defined as disability at the time of discharge or prolongation of the hospitalization caused by medical treatment as opposed to the disease process, were assessed through review of approximately 31,000 hospital records in New York in 1984. These sampled records were then linked to formal malpractice claims. To estimate the risk of malpractice claims by age, gender, race, insurance status, and income, we conducted a case-control study of claimant cases matched with nonclaimant controls. The cases were all those patients who filed malpractice claims referring to alleged malpractice during a sampled hospitalization. Physician reviewers had previously judged the level of disability that resulted from the medical injury. Claimants (n = 51) were each matched with five nonclaimant controls on the basis of injury. Noninjured cases were matched with noninjured controls and injured cases were matched with injured controls. RESULTS: We found that poor patients (odds ratio [OR], 0.2; 95% confidence interval [CI], 0.03 to 0.8) and uninsured patients (OR, 0.1; 95% CI, 0.005 to 0.9) were significantly less likely to file malpractice claims, after controlling for the severity of medical injury. Among patients who suffered medical injury, the elderly (OR, 0.2; 95% CI, 0.03 to 0.9) were also less likely to file claims. Gender and race were not independently associated with risk of malpractice claims. CONCLUSIONS: Poor and uninsured patients are significantly less likely to sue for malpractice, even after controlling for the presence of medical injury. Fear of malpractice risk should not be a significant factor in the decision to serve the poor. Tort reforms that would protect physicians who serve the medically indigent from malpractice suits may not be warranted. PMID- 8411500 TI - Prediction of mortality and morbidity with a 6-minute walk test in patients with left ventricular dysfunction. SOLVD Investigators. AB - OBJECTIVE: To study the potential usefulness of the 6-minute walk test, a self paced submaximal exercise test, as a prognostic indicator in patients with left ventricular dysfunction. DESIGN: Data were collected during a prospective cohort study, the Studies of Left Ventricular Dysfunction (SOLVD) Registry Substudy. SETTING: Twenty tertiary care hospitals in the United States, Canada, and Belgium. PARTICIPANTS: A stratified random sample of 898 patients from the SOLVD Registry who had either radiological evidence of congestive heart failure and/or an ejection fraction of 0.45 or less were enrolled in the substudy and underwent a detailed clinical evaluation including a 6-minute walk test. Patients were followed up for a mean of 242 days. OUTCOME MEASURES: Mortality and hospitalization. RESULTS: During follow-up, 52 walk-test participants (6.2%) died and 252 (30.3%) were hospitalized. Hospitalization for congestive heart failure occurred in 78 participants (9.4%), and the combined endpoint of death or hospitalization for congestive heart failure occurred in 114 walk-test participants (13.7%). Compared with the highest performance level, patients in the lowest performance level had a significantly greater chance of dying (10.23% vs 2.99%; P = .01), of being hospitalized (40.91% vs 19.90%; P = .002), and of being hospitalized for heart failure (22.16% vs 1.99%; P < .0001). In a logistic regression model, ejection fraction and distance walked were equally strong and independent predictors of mortality and heart failure hospitalization rates during follow-up. CONCLUSION: The 6-minute walk test is a safe and simple clinical tool that strongly and independently predicts morbidity and mortality in patients with left ventricular dysfunction. PMID- 8411501 TI - Impact of first-responder defibrillation in an urban emergency medical services system. AB - OBJECTIVE: To evaluate the impact of adding first-responder defibrillation by fire-fighters to an existing advanced life-support emergency medical services system. DESIGN: Nonrandomized, controlled clinical trial with periodic crossover. SETTING: Memphis, Tenn, a city of 610,337 people, which is served by a fire department-based emergency medical services system. All city ambulances provide advanced life support. PATIENTS: Adult victims of out-of-hospital cardiac arrest due to heart disease. INTERVENTION: Twenty of 40 participating engine companies were equipped with an automated external defibrillator and ordered to apply it immediately in all cases of cardiac arrest. The other 20 companies were ordered to start cardiopulmonary resuscitation (CPR) immediately and wait for paramedics to arrive. Every 75 days, group roles were reversed. Care otherwise proceeded according to 1986 American Heart Association guidelines. MAIN OUTCOME MEASURES: Return of spontaneous circulation in the field, survival to hospital admission, survival to hospital discharge, and neurological status at discharge. RESULTS: During the 39-month study interval, 879 patients were treated by a project engine company. Four hundred thirty-one (49%) of these were found in ventricular fibrillation. Bystander CPR was started in only 12% of cases. Overall, firefighters reached the scene a mean of 2.5 minutes faster than simultaneously dispatched paramedics. Although our automated external defibrillators proved to be reliable and efficacious for terminating ventricular fibrillation and pulseless ventricular tachycardia, patients treated by an automated external defibrillator-equipped engine company were no more likely than CPR-treated controls to be resuscitated (32% vs 34%, respectively), to survive to hospital admission (31% vs 29%), or to survive to hospital discharge (14% vs 10%). Neurological outcomes were also similar in the two treatment groups. CONCLUSIONS: In a fast-response, urban emergency medical services system served by paramedics, the impact of adding first-responder defibrillation appears to be small. Early defibrillation alone cannot overcome low community rates of bystander CPR. Careful attention to every link in the "chain of survival" is needed to achieve optimal rates of survival after cardiac arrest. PMID- 8411502 TI - Ophthalmic examination among adults with diagnosed diabetes mellitus. AB - OBJECTIVE: To assess whether adults with diagnosed diabetes in the United States are receiving recommended eye examinations for detection of diabetic retinopathy and what factors are associated with receiving them. DESIGN, SETTING, AND PARTICIPANTS: The design was a cross-sectional survey of the civilian, noninstitutionalized US population 18 years of age or older, based on the 1989 National Health Interview Survey. A multistage probability sampling strategy was used to identify a representative sample of 84,572 persons. A questionnaire on diabetes was administered to all subjects with diagnosed diabetes (n = 2405). MAIN OUTCOME MEASURE: A dilated eye examination in the past year. MAIN RESULTS: Of all adults with diagnosed diabetes in the United States, only 49% had a dilated eye examination in the past year. This included 57% of people with insulin-dependent diabetes mellitus (IDDM), 55% with insulin-treated non-insulin dependent diabetes mellitus (NIDDM), and 44% with NIDDM not treated with insulin. Even among diabetics at high risk of vision loss because of retinopathy or long duration of diabetes, the proportion with a dilated eye examination was only 61% and 57%, respectively. By logistic regression, the probability of a dilated eye examination among persons with NIDDM increased with older age, higher socioeconomic status, and having attended a diabetes education class. The probability of a dilated eye examination was not independently related to race, duration of diabetes, frequency of physician visits for diabetes, or health insurance. CONCLUSIONS: About half of adults with diabetes in the United States are not receiving timely and recommended eye care to detect and treat retinopathy. Widespread interventions, including patient and professional education, are needed to ensure that diabetic patients who are not receiving appropriate eye care have an annual dilated eye examination to detect retinopathy and prevent vision loss. PMID- 8411503 TI - Immunoaugmentative therapy. An unproven cancer treatment. AB - While it is statistically true that clinical cancer occasionally occurs following development of immune incompetency, the fact remains that the most common forms of cancer (ie, breast, lung, and colon) are not experienced by immunosuppressed individuals. Since there is strong scientific evidence that the unprovoked, normal immune system does not recognize and destroy cancer cells that arise spontaneously, the concept of an immune surveillance system that continuously protects clinically normal humans from cancer remains an appealing but unproven hypothesis. Burton's theory of immune surveillance against cancer appears to be nothing more than a rehash of the hypotheses of many other investigators, embellished with his postulate that four specific anticancer protein factors exist and function in the normal human immune system. The IAT he invented as a treatment for cancer is based on his presumption that he has proven the existence of these factors. But neither his declarations nor those of his proponents offer any objective evidence to support such a conclusion. They have not proven that the IAT components exist or can be extracted from blood without the loss of biologic activity. The tests that are described cannot quantitatively measure the specific IAT proteins and there is no evidence that the IAT fractions possess any immunologic activity. Thus, Burton's tumor antibody has never been shown to be a tumor specific immunoglobulin that can interact with tumor antigens and activate complement. His tumor complement fraction has no complement activity (written communication, G.J. Gray, PhD, June 21, 1984), his blocking protein has not been shown to block anything, his deblocking protein has not been shown to deblock anything, and his human tumor cells have never been shown to lyse following the interaction of tumor antibody and tumor complement. Finally, since there is no information on the quality control procedures being used in the manufacture of the IAT materials, there is the possibility that they may be unsterile and, therefore, hazardous for use in humans. Patients who are considering IAT as treatment for cancer should be made aware of these facts when attempting to reach an informed decision regarding its safety and potential efficacy. While this paper was being reviewed for publication, an IAT proponent newsletter called The Cancer Chronicles published the news that Lawrence Burton died of a heart attack in March 1993. The editor of this newsletter, Ralph Moss, PhD, stated that Burton's clinic would remain open under the direction of its medical director, R. John Clement, MD, and would continue to offer IAT to cancer patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8411504 TI - Identification of the critical oxygen delivery for anaerobic metabolism in critically ill septic and nonseptic humans. AB - OBJECTIVES: To determine the critical oxygen delivery threshold for anaerobic metabolism and to compare its value between septic and nonseptic critically ill patients. DESIGN: Cohort analytic study, consecutive sample. SETTING: Two tertiary care medical and surgical intensive care units in university hospitals. PATIENTS: Nine septic and nine nonseptic critically ill humans. A diagnosis of sepsis was established by the presence of sepsis syndrome, positive cultures obtained within 48 hours of study, and autopsy evidence of a source of infection. METHODS AND INTERVENTIONS: The O2 consumption (determined by indirect calorimetry), O2 delivery (calculated from the Fick equation), and concentration of arterial plasma lactate were simultaneously determined at 5- to 20-minute intervals while life support was discontinued. MAIN OUTCOME MEASURES: Critical O2 delivery, critical O2 extraction ratio, and maximal O2 extraction ratio. RESULTS: In all septic and eight nonseptic patients, O2 delivery and O2 consumption displayed a biphasic relationship over the range of O2 delivery studied. There were no differences in critical O2 delivery threshold (3.8 +/- 1.5 vs 4.5 +/- 1.3 mL.min-1 x kg-1; P > .28), critical O2 extraction ratio (0.61 +/- 0.05 vs 0.59 +/ 0.16; P > .64), and maximal O2 extraction ratio (0.74 +/- 0.08 vs 0.80 +/- 0.11; P > .29) between septic and nonseptic patients. These data have greater than 90% power to detect a difference of 2 mL.min-1 x kg-1 in the critical O2 delivery and 0.1 in the critical and maximal O2 extraction ratios between the septic and nonseptic groups. CONCLUSIONS: The critical O2 delivery for anaerobic metabolism was identified from the biphasic relationship between O2 delivery and O2 consumption in individual humans. The critical O2 delivery is considerably lower than previously reported in humans with the use of pooled group data. Sepsis does not alter the critical O2 delivery for anaerobic metabolism or tissue O2 extraction ability. Interventions to increase O2 delivery to supranormal levels in critically ill humans in the hope of increasing O2 consumption may be inappropriate. PMID- 8411505 TI - Percutaneous mitral balloon valvotomy and the new demographics of mitral stenosis. AB - OBJECTIVE: This review discusses the latest developments in selected clinical features and catheter-based therapy of mitral stenosis. DATA SOURCES: English language journal articles and reviews in the clinical and epidemiological literature as related to mitral valve stenosis from 1965 through March 1993, identified by bibliography review and expert consultation. STUDY SELECTION: Selected studies included clinical trials with adequate patient population description and short- and long-term (5 years) follow-up for topics related to mitral valve stenosis in the clinical literature. DATA EXTRACTION: Two reviewers participated in extracting the data with the aim of presenting a balanced and comprehensive review of the subject. DATA SYNTHESIS AND CONCLUSIONS: The main conclusions are (1) mitral stenosis should no longer be viewed as a largely "geriatric disease" in the United States due to a recent inflow many young immigrants from countries where rheumatic fever continues partially or wholly unabated; (2) clinical and anatomical features of mitral stenosis are age dependent; when clinical presentation occurs at 30 vs 70 years of age, for example, the degree of valve obstruction may be similar but differences exist in the frequency of atrial fibrillation, the magnitude of reduction in cardiac output, the degree of valve deformity and calcification, and the frequency of coexistent coronary artery disease; and (3) mitral stenosis therapy has undergone a reorientation with the introduction of percutaneous mitral balloon valvotomy, which has proven to be safe, cost-effective, and to provide short- and long-term improvements in symptomatic and hemodynamic status in selected patients. PMID- 8411506 TI - Two wrongs don't make a right. Managed care, mental health, and the marketplace. PMID- 8411507 TI - Caring for the poor and professional liability. Is there a need for tort reform? PMID- 8411508 TI - Christian Science church loses first civil suit on wrongful death of a child. PMID- 8411509 TI - Major changes seen for US childhood vaccination. PMID- 8411510 TI - Infant AIDS prevention study finally gets going. PMID- 8411511 TI - From the Centers for Disease Control and Prevention. Update: influenza activity- United States and worldwide, 1993. PMID- 8411512 TI - From the Centers for Disease Control and Prevention. Mammography and clinical breast examinations among women aged 50 years and older--Behavioral Risk Factor Surveillance System, 1992. PMID- 8411513 TI - From the Centers for Disease Control and Prevention. Update: National Breast and Cervical Cancer Early Detection Program, 1992-1993. PMID- 8411514 TI - From the Centers for Disease Control and Prevention. Nosocomial enterococci resistant to vancomycin--United States, 1989-1993. PMID- 8411515 TI - Noninvasive testing for silent myocardial ischemia in stable coronary patients. PMID- 8411516 TI - Noninvasive testing for silent myocardial ischemia in stable coronary patients. PMID- 8411517 TI - Mechanical ventilation for the elderly. PMID- 8411518 TI - Percutaneous transluminal coronary angioplasty in New York State. PMID- 8411519 TI - Percutaneous transluminal coronary angioplasty in New York State. PMID- 8411520 TI - Over-the-counter cold remedies. PMID- 8411521 TI - Why estimates of physician supply and requirements disagree. PMID- 8411522 TI - Why estimates of physician supply and requirements disagree. PMID- 8411523 TI - Why estimates of physician supply and requirements disagree. PMID- 8411524 TI - A piece of my mind. The rest of him. PMID- 8411525 TI - Depression following myocardial infarction. Impact on 6-month survival. AB - OBJECTIVE: To determine if the diagnosis of major depression in patients hospitalized following myocardial infarction (MI) would have an independent impact on cardiac mortality over the first 6 months after discharge. DESIGN: Prospective evaluation of the impact of depression assessed using a modified version of the National Institute of Mental Health Diagnostic Interview Schedule for major depressive episode. Cox proportional hazards regression was used to evaluate the independent impact of depression after control for significant clinical predictors in the data set. SETTING: A large, university-affiliated hospital specializing in cardiac care, located in Montreal, Quebec. PATIENTS: All consenting patients (N = 222) who met established criteria for MI between August 1991 and July 1992 and who survived to be discharged from the hospital. Patients were interviewed between 5 and 15 days following the MI and were followed up for 6 months. There were no age limits (range, 24 to 88 years; mean, 60 years). The sample was 78% male. PRIMARY OUTCOME MEASURE: Survival status at 6 months. RESULTS: By 6 months, 12 patients had died. All deaths were due to cardiac causes. Depression was a significant predictor of mortality (hazard ratio, 5.74; 95% confidence interval, 4.61 to 6.87; P = .0006). The impact of depression remained after control for left ventricular dysfunction (Killip class) and previous MI, the multivariate significant predictors of mortality in the data set (adjusted hazard ratio, 4.29; 95% confidence interval, 3.14 to 5.44; P = .013). CONCLUSION: Major depression in patients hospitalized following an MI is an independent risk factor for mortality at 6 months. Its impact is at least equivalent to that of left ventricular dysfunction (Killip class) and history of previous MI. Additional study is needed to determine whether treatment of depression can influence post-MI survival and to assess possible underlying mechanisms. PMID- 8411526 TI - Pneumococcal polysaccharide vaccine efficacy. An evaluation of current recommendations. AB - OBJECTIVE: To determine pneumococcal polysaccharide vaccine efficacy in selected populations at risk for serious pneumococcal infection for whom vaccination is currently recommended and to assess duration of protection after vaccination. DESIGN: Vaccine efficacy was estimated using indirect cohort analysis to compare the proportion of pneumococcal infections caused by serotypes included in the vaccines of vaccinated and unvaccinated persons who were identified during 14 years of national surveillance. SETTING: Hospital laboratories in the United States that submitted pneumococcal isolates to the Centers for Disease Control and Prevention between May 1978 and April 1992. PARTICIPANTS: A total of 2837 persons older than 5 years who had pneumococcus isolated from blood or cerebrospinal fluid. RESULTS: Overall efficacy for preventing infection caused by serotypes included in the vaccine was 57% (95% confidence interval [CI], 45% to 66%). Efficacy among persons with diabetes mellitus was 84% (95% CI, 50% to 95%); with coronary vascular disease, 73% (95% CI, 23% to 90%); with congestive heart failure, 69% (95% CI, 17% to 88%); with chronic pulmonary diseases, 65% (95% CI, 26% to 83%); and with anatomic asplenia, 77% (95% CI, 14% to 95%). Efficacy was not documented for patients with alcoholism or cirrhosis, sickle cell disease, chronic renal failure, lymphoma, leukemia, or multiple myeloma, although sample sizes were small for these groups. Efficacy for immunocompetent persons older than 65 years was 75% (95% CI, 57% to 85%). Efficacy did not decline with increasing interval after vaccination: 5 to 8 years after vaccination it was 71% (95% CI, 24% to 89%), and 9 years or more after vaccination it was 80% (95% CI, 16% to 95%). CONCLUSIONS: Intensified efforts to improve pneumococcal vaccine coverage among certain populations for whom vaccination is currently recommended is indicated, but universal revaccination is not warranted at this time. PMID- 8411527 TI - Temporal changes in the care and outcomes of elderly patients with acute myocardial infarction, 1987 through 1990. AB - OBJECTIVE: To investigate changes between 1987 and 1990 in the care and outcomes associated with acute myocardial infarction (AMI) in elderly patients. DESIGN: Retrospective cohort study using a longitudinal database created from Medicare administrative files. PATIENTS: Cohorts comprising a total of 856,847 AMI patients insured by Medicare between 1987 and 1990. MAIN OUTCOME MEASURES: Annual rates of mortality at 30 days and 1 year following AMI, and the use of coronary angiography, coronary artery bypass graft surgery, and percutaneous transluminal coronary angioplasty during the first 90 days after a new AMI. RESULTS: Between 1987 and 1990, mortality rates decreased 10% overall from 26% to 23% at 30 days (P < .001) and from 40% to 36% at 1 year following AMI (P < .001). Declines in mortality and adjusted risks of 1-year mortality were similar in men and women and in blacks and whites, but mortality declines were more evident in those younger than 85 years. Meanwhile, the proportion of elderly AMI patients having angiography within the first 90 days after their index admission increased from 24% to 33% (P < .001); proportions increased for both genders and all races. The proportion of patients undergoing revascularization procedures increased from 13% to 21%; while rates of bypass surgery increased from 8% to 11%, rates of angioplasty doubled from 5% to 10% (all P < .001). CONCLUSIONS: Between 1987 and 1990, survival of elderly patients following AMI improved significantly. While changes in patient treatment may be responsible, the increased use of thrombolytic therapy appears to be only a partial explanation. Also, while the use of coronary angiography and revascularization procedures increased dramatically, the degree to which it caused the improvement in survival could not be determined. PMID- 8411528 TI - Myositis-specific autoantibodies. Touchstones for understanding the inflammatory myopathies. PMID- 8411529 TI - Adolescents as victims of family violence. Council on Scientific Affairs, American Medical Association. AB - Adolescents experience maltreatment at rates equal to or exceeding those of younger children. Recent increases in reported cases of maltreatment have occurred disproportionately among older children and adolescents. However, adolescents are less likely to be reported to child protective services and are more likely to be perceived as responsible for their maltreatment. Adolescent girls are reported as victims more often than boys, especially in sexual abuse. However, boys may be less likely to be identified or reported and often are abused by nonfamily members. Parents of adolescent victims have higher average income and educational levels and are less likely to have a parental history of abuse than parents of younger children. A wide range of serious adolescent risk behaviors is associated with maltreatment. These include increased risk of premature sexual activity, unintended pregnancy, emotional disorders, suicide attempts, eating disorders, alcohol and other drug abuse, and delinquent behavior. Incarcerated youth, homeless or runaway youth, and youth who victimize siblings or assault parents have been shown to have high rates of prior maltreatment. Signs of maltreatment are often ambiguous for adolescents. Screening questions have been effective in prompting self-disclosure of abuse. Adolescents also experience problems in the child welfare system that offers fewer and less appropriate services for this age group. Recommendations are made regarding screening of adolescents for maltreatment, the development of better services for adolescents, research on parenting to prevent maltreatment, and training of school staff to identify and refer victims of maltreatment. PMID- 8411530 TI - A world without polio. 'Future generations will know by history only...'. PMID- 8411531 TI - Psychosocial factors and prognosis in established coronary artery disease. The need for research on interventions. PMID- 8411532 TI - Scapulothoracic pain. PMID- 8411533 TI - Vena caval filters and anticoagulants for pulmonary emboli. PMID- 8411534 TI - Aluminum in Alzheimer's disease. PMID- 8411535 TI - New Federal stances on TB control may be confusing to health care facilities. PMID- 8411536 TI - Major players in unique new huddle with goal of developing better strategies for AIDS research. PMID- 8411537 TI - From the Institute of Medicine. PMID- 8411538 TI - From the Centers for Disease Control and Prevention. Alcohol involvement in pedestrian fatalities--United States, 1982-1992. PMID- 8411539 TI - From the Centers for Disease Control and Prevention. Prevalence of mobility and self-care disability--United States, 1990. PMID- 8411540 TI - From the Centers for Disease Control and Prevention. Update: hantavirus associated illness--North Dakota, 1993. PMID- 8411541 TI - From the Centers for Disease Control and Prevention. Progress in the development of hantavirus diagnostic assays--United States. PMID- 8411542 TI - From the Centers for Disease Control and Prevention. Prevalence of work disability--United States, 1990. PMID- 8411543 TI - Voluntary organ donation: autonomy ... tragedy. PMID- 8411544 TI - Faculty practice plans: caring for the underserved? PMID- 8411545 TI - Childhood vaccinations: are rates really declining? PMID- 8411546 TI - Canadian Navy dampens smokers' habit. PMID- 8411547 TI - Panculture panned. PMID- 8411548 TI - Azithromycin in chlamydial urethritis. PMID- 8411549 TI - Relax: it's onlie sodeyne death. PMID- 8411550 TI - Examining product risk in context. Market withdrawal of zomepirac as a case study. AB - OBJECTIVE: To examine changes in the prescribing of analgesics after the market entry and subsequent withdrawal of zomepirac sodium, a nonsteroidal anti inflammatory drug (NSAID), following repeated reports of zomepirac-related deaths. DESIGN: To evaluate this natural quasi experiment, we conducted time series analyses to compare prescribing in two cohorts of primary care physicians from July 1980 through September 1983. SETTING: Study physicians provided outpatient pharmaceutical care to patients enrolled in the New Jersey Medicaid program. PARTICIPANTS: We identified 260 primary care physicians who provided 10 or more prescriptions for zomepirac (zomepirac prescribers) and 308 who provided 10 or more prescriptions for NSAIDs other than zomepirac (other-NSAID prescribers) in Medicaid during the study period. MAIN OUTCOME MEASURES: Monthly rates of prescribing for zomepirac and several categories of substitute analgesics among Medicaid patients seen by study physicians. MAIN RESULTS: Zomepirac accounted for a stable 11.0% of analgesic prescribing among the zomepirac-prescriber cohort; label changes and manufacturer product-risk warnings 11 months before the product's withdrawal from the market had no impact on use. After market entry, zomepirac prescribers reduced use of other NSAIDs and propoxyphene (hydrochloride or napsylate) in comparison with other-NSAID prescribers (-8.1% and -2.8% of total analgesic prescribing, respectively; P < .001). After the product's withdrawal from the market, zomepirac prescribers showed significant increases in relative prescribing of other NSAIDs (+6.8%; P < .001), propoxyphene (+2.1%; P < .05), and analgesics containing barbiturates (+2.7%; P < .001). CONCLUSIONS: The sudden withdrawal of zomepirac from the market resulted in substitutions not only of other NSAIDs, but also of alternative analgesics that carry risks of habituation and adverse effects. Apparent gains in patient safety resulting from market withdrawal of medications must be evaluated in comparison with risks of medications likely to be substituted. PMID- 8411551 TI - Accuracy and reproducibility of precordial percussion and palpation for detecting increased left ventricular end-diastolic volume and mass. A comparison of physical findings and ultrafast computed tomography of the heart. AB - OBJECTIVE: To assess the accuracy and reproducibility of indirect definitive precordial percussion in detecting increased left ventricular end-diastolic volume (LVEDV), left ventricular mass (LVM), and left ventricular end-diastolic wall thickness (LVEDWT), and to compare it with palpation of the apical impulse. DESIGN: Descriptive study. SETTING: Hospitals and clinics of a university medical center. PATIENTS: Convenience sample of 103 patients (62 men and 41 women) referred for ultrafast computed tomography (CT) of the heart. INTERVENTIONS: Percussion dullness distance from the midsternal line in the left fourth through sixth intercostal spaces, distance of the apical impulse from the midsternal line, and apical impulse diameter in the left lateral decubitus position were measured on all patients. Measurements of LVEDV, LVM, and LVEDWT were taken using ultrafast CT of the heart. Investigators performing the physical diagnostic maneuvers were blinded to the clinical history and CT results, and investigators performing the CT scans were blinded to physical findings. RESULTS: Percussion dullness distance in the left fifth intercostal space was the best discriminator of LVEDV (receiver operating characteristic [ROC] area, 0.680; 95% confidence interval [CI], 0.547 to 0.813), and dullness distance in the left sixth intercostal space was the best discriminator of LVM and LVEDWT (ROC areas, 0.831, 95% CI, 0.674 to 0.988; and 0.849, 95% CI, 0.651 to 0.999, respectively). A percussion dullness distance of greater than 10.5 cm in the left fifth intercostal space detected increased LVEDV or LVM with a sensitivity of 91.3% (95% CI, 70.5% to 98.5%) and a specificity of 30.3% (95% CI, 19.9% to 43.0%). There was moderate concordance between investigators for percussion dullness distance (kappa, 0.57; 95% CI, 0.18 to 0.96). In patients in whom an impulse was palpated, an apical impulse diameter of greater than 3.0 cm in the left lateral decubitus detected increased LVEDV or LVM with a sensitivity of 100% (95% CI, 77.1% to 100%) and a specificity of 40% (95% CI, 23.2% to 59.3%). However, an impulse was palpable in only 53% of cases and showed only slight interobserver reproducibility (kappa, 0.18; 95% CI, 0.0 to 0.58). CONCLUSION: Indirect definitive percussion of the precordium is a sensitive and moderately reproducible maneuver for excluding cardiomegaly due to increased LVEDV or LVM. Although measurement of apical impulse diameter was also sensitive in excluding cardiomegaly, lack of a palpable impulse in many patients and low precision between physicians may limit its utility in clinical practice. PMID- 8411552 TI - Factors affecting late mortality from heart disease after treatment of Hodgkin's disease. AB - OBJECTIVE: To assess the risk of death from heart disease after Hodgkin's disease therapy. DESIGN: Retrospective study comparing treated patients with a matched general population. SETTING: Referral center. PATIENTS: A total of 2232 consecutive Hodgkin's disease patients treated from 1960 through 1991. Follow-up averaged 9.5 years. MAIN OUTCOME MEASURES: Relative risks (RRs), the ratio of the observed to the expected cases with 95% confidence intervals (CIs), chi tests for trends, and Kaplan-Meier actuarial risks. RESULTS: Of the 2232 patients, 88 (3.9%) died of heart disease, 55 from acute myocardial infarction and 33 from other cardiac diseases, including congestive heart failure, radiation pericarditis or pancarditis, cardiomyopathy, or valvular heart disease. The RR for cardiac death was 3.1 (CI, 2.4 to 3.7). Mediastinal radiation of 30 Gy or less (n = 385 patients) did not increase risk; above 30 Gy (n = 1830), RR was 3.5 (CI, 2.7 to 4.3). Blocking to limit cardiac exposure reduced the RR for other cardiac diseases from 5.3 (CI, 3.1 to 7.5) to 1.4 (CI, 0.6 to 2.9), but not acute myocardial infarction (RR, 3.7 vs 3.4). The RRs increased with duration after treatment (trend in acute myocardial infarction, P = .02; in other cardiac diseases, P = .004). The RR for acute myocardial infarction was highest after irradiation before 20 years of age and decreased with increasing age at treatment (P < .0001 for trend). CONCLUSIONS: Mediastinal irradiation for Hodgkin's disease increases the risk of subsequent death from heart disease. Risk increased with high mediastinal doses, minimal protective cardiac blocking, young age at irradiation, and increasing duration of follow-up. PMID- 8411553 TI - Clinical effectiveness of influenza vaccination in Manitoba. AB - OBJECTIVE: To assess the clinical effectiveness of influenza vaccination in preventing influenza-associated hospitalization and death. DESIGN: Case-control study. SETTING AND PATIENTS: Noninstitutionalized persons aged 45 years or older living in Manitoba, on December 1, 1982, and December 1, 1985. METHODS: Linked records of the Manitoba population registry, hospital-discharge abstracts, physician claims for ambulatory-patient visits and influenza vaccination, and vital statistics were used. A matched-set analysis estimated the clinical effectiveness of influenza vaccination in preventing hospital admissions and deaths from influenza-associated conditions during influenza A (H3N2) outbreak periods in 1982 to 1983 (12 weeks) and 1985 to 1986 (10 weeks). The analysis adjusted for hospital discharge and ambulatory care for high-risk conditions within the previous 15 months and 3 months, respectively. RESULTS: Influenza vaccination prevented 32% to 39% of hospital admissions with pneumonia and influenza and 15% to 34% of admissions with all respiratory conditions. Vaccination was 43% to 65% effective in preventing hospital deaths with these conditions (all listed diagnoses) and 27% to 30% effective in preventing deaths from all causes. CONCLUSION: Influenza vaccination has substantial clinical effectiveness in preventing hospital admission and death from influenza associated conditions in noninstitutionalized individuals. PMID- 8411554 TI - Ventilator-associated pneumonia. A multivariate analysis. AB - OBJECTIVES: To identify factors associated with the development of ventilator associated pneumonia (VAP) and to examine the incidence of VAP in different intensive care unit (ICU) populations. DESIGN: An inception cohort study. SETTING: Barnes Hospital, St Louis, Mo, an academic tertiary care center. PATIENTS OR OTHER PARTICIPANTS: A total of 277 consecutive patients required mechanical ventilation for longer than 24 hours from a medical ICU (75 patients), surgical ICU (100 patients), or cardiothoracic ICU (102 patients). INTERVENTIONS: Prospective patient surveillance and data collection. MAIN OUTCOME MEASURES: Ventilator-associated pneumonia and ICU mortality. RESULTS: Ventilator-associated pneumonia occurred in 43 patients (15.5%). Stepwise logistic regression analysis identified four factors to be independently associated with VAP (P < .05): an organ system failure index of 3 or greater (adjusted odds ratio [AOR] = 10.2; 95% confidence interval [CI], 4.5 to 23; P < .001); patient age of 60 years or older (AOR = 5.1; 95% CI, 1.9 to 14.1; P = .002); prior administration of antibiotics (AOR = 3.1; 95% CI, 1.4 to 6.9; P = .004); and supine head positioning during the first 24 hours of mechanical ventilation (AOR = 2.9; 95% CI, 1.3 to 6.8; P = .013). Ventilator-associated pneumonia occurred more often in cardiothoracic patients (21.6%) compared with medical patients (9.3%) (P = .03). Patients with VAP also had a higher mortality (37.2%) than those without VAP (8.5%) (P < .001). An organ system failure index of 3 or greater (AOR = 16.1; 95% CI, 6.1 to 42; P < .001), a premorbid lifestyle score of 2 or greater (AOR = 3.1; 95% CI, 1.3 to 7.3; P = .012), and supine head positioning during the first 24 hours of mechanical ventilation (AOR = 3.1; 95% CI, 1.2 to 7.8; P = .016) were independently associated with mortality. CONCLUSIONS: These data suggest potential interventions that might affect the incidence of VAP or outcome associated with VAP. Additionally, they indicate that different ICU populations may have different incidences of VAP. PMID- 8411555 TI - A study of various tests to detect asymptomatic urinary tract infections in an obstetric population. AB - OBJECTIVE: To compare rapid screening techniques for detecting asymptomatic urinary tract infections (AUTIs) in pregnant women. DESIGN: Comparison of results of the screening tests of urinalysis, urine dipstick, and Gram's staining with the results of standard urine culture at an initial prenatal visit. In follow-up visits, urine dipstick testing was compared with urinalysis. SETTING: Departments of Family Medicine and Obstetrics and Gynecology, Mayo Clinic, Rochester, Minn. PATIENTS: Pregnant women (1047) from the local community were screened for AUTI on initial and follow-up visits. METHODS: Initial prenatal urine was tested by using urine dipstick testing, urinalysis, Gram's staining, and urine culture. At each follow-up visit, urine specimens were tested by using urine dipstick and urinalysis. MAIN OUTCOME MEASURES: Sensitivity and specificity, incremental patient costs, and clinical outcomes were used to assess the effectiveness of the techniques. RESULTS: On initial visits, rapid screening tests for AUTI in pregnant women revealed the following: Gram's staining identified 22 of 24 patients with AUTI (sensitivity, 91.7%; specificity, 89.2%); urine dipstick, 12 of 24 (sensitivity, 50.0%; specificity, 96.9%); and urinalysis with presence of leukocytes, six of 24 (sensitivity, 25.0%; specificity, 99.0%). In follow-up visits, urine dipstick tests detected 19 infections and urinalysis, three (positive predictive value, 5% compared with 3%). CONCLUSIONS: Urine dipstick testing for nitrites identified half of all patients with urinary tract infections and was superior to urinalysis on follow-up visits. Although Gram's staining is more expensive, it was more accurate for AUTI than urinalysis or urine dipstick test for nitrites. Urinalysis was never the test of choice because it was more expensive and detected fewer positive cultures. Leukocyte measurement correlated poorly with AUTI. PMID- 8411556 TI - A view from out there. A second look. PMID- 8411557 TI - Withdrawal of a drug from the market. What should the prescriber do? PMID- 8411558 TI - Kicking butts--AMA, Joe Camel, and the 'black-flag' war on tobacco. PMID- 8411559 TI - Fast-track improves CABG outcomes. PMID- 8411560 TI - Radical approaches: is widespread testing and treatment for oxidative injuries coming soon? PMID- 8411561 TI - Coming to terms with indications for fetal surgery. PMID- 8411562 TI - From the Congressional Office of Technology Assessment. PMID- 8411563 TI - From the Centers for Disease Control and Prevention. Violence-related attitudes and behaviors of high school students--New York City, 1992. PMID- 8411564 TI - From the Centers for Disease Control and Prevention. Self-reported HIV-antibody testing among persons with selected risk behaviors--southern Los Angeles County, 1991-1992. PMID- 8411565 TI - From the Centers for Disease Control and Prevention. Unintentional carbon monoxide poisoning from indoor use of pressure washers--Iowa, January 1992 January 1993. PMID- 8411566 TI - Metered-dose inhalers: drug delivery over the life of a canister. PMID- 8411567 TI - Do we have yet another spotted fever? PMID- 8411568 TI - Hawaiian health plan: less than meets the eye? PMID- 8411569 TI - Hawaiian health plan: less than meets the eye? PMID- 8411570 TI - No bells! No whistles! A cognocardiologist! PMID- 8411571 TI - Costs and benefits of lead screening. PMID- 8411572 TI - An economic evaluation of screening for Chlamydia trachomatis in adolescent males. AB - OBJECTIVE: To assess the cost-effectiveness of identifying asymptomatic carriers of Chlamydia trachomatis among adolescent males. DESIGN: Cost-effectiveness analysis based on cohort analytic studies previously reported and average salaries and costs of medical care in Sweden. SETTING: Adolescent males attending a primary care center for routine health checks. PARTICIPANTS: Estimates of costs and benefits are based on a cohort of 1000 adolescent males and their female contacts. INTERVENTION: Screening with enzyme immunoassay (EIA), either on leukocyte esterase (LE)--positive urine samples (LE-EIA screening) or on all urine samples (EIA screening), was compared with no screening (no treatment or contact tracing). The effects of confirming positive EIA results with a blocking assay and alternative antibiotic regimens on the outcome of the screening strategies were also evaluated. RESULTS: Compared with no screening, the LE-EIA and EIA screening strategies reduced the overall costs when the prevalence of chlamydial infection in males exceeded 2% and 10%, respectively. Enzyme immunoassay screening achieved an overall cure rate that was 12.2% to 12.6% (95% confidence interval) better, but reduced the incremental savings by at least $2144 per cured male, in comparison with LE-EIA screening. Confirmation of positive EIA results reduced the overall cost of the LE-EIA screening strategy when the prevalence of C trachomatis among males was less than 8%. Compared with a 7-day course of doxycycline, a single oral dose of azithromycin administered under supervision in the clinic improved the cure rates of both EIA and LE-EIA screening strategies by 15.1% to 16.3% and 11.2% to 12.0%, respectively, while reducing the corresponding overall costs by 5% and 9%, respectively, regardless of the prevalence of chlamydial infection in males. CONCLUSION: The use of LE-EIA screening combined with treatment of positive cases with azithromycin was the most cost-effective intervention strategy focusing on asymptomatic male carriers of C trachomatis. Positive EIA results should be confirmed when screening low risk populations. PMID- 8411573 TI - Evaluation of urine-based screening strategies to detect Chlamydia trachomatis among sexually active asymptomatic young males. AB - OBJECTIVE: To evaluate the performances of diagnostic screening tests alone or in combination to detect asymptomatic chlamydial urethral infection in young males. DESIGN: Comparisons of the performance profiles of the following chlamydia screening strategies were done: urethral culture; identification of polymorphonucleocytes (PMNs) on spun first-void urine (FVU); urinary leukocyte esterase test (LET) on unspun FVU; chlamydial enzyme immunoassay (EIA) applied to FVU sediment; combining LET on unspun FVU followed by EIA with or without direct fluorescent antibody (DFA) confirmation on FVU sediment; and combining PMNs on spun FVU followed by EIA with or without DFA confirmation. SETTING: General clinics at a youth detention center, university-based teen clinic, college health service, and a military screening clinic. PATIENTS: A total of 618 males aged 12 to 35 years (mean, 17 years) were recruited as a convenience sample; site participation rates ranged from 50% to 80%. Eligible subjects were sexually active, denied symptoms of urethritis, and had taken no antibiotics in the prior 2 weeks. MAIN OUTCOME MEASURES: Sensitivity, specificity, and positive and negative predictive values of each test strategy's ability to detect Chlamydia trachomatis infection, and cost to confirm each positive case. RESULTS: With a 7% prevalence of chlamydial infection, tissue culture had a sensitivity of only 61%. However, two strategies yielded significantly better performance profiles compared with the others: EIA confirmed by DFA test with a sensitivity of 84%, a specificity of 100%, and a cost to identify each positive case of $434; and PMNs followed by EIA confirmed by DFA test with a sensitivity of 78%, a specificity of 100%, and a cost to identify each positive case of $199. The LET followed by EIA DFA had a similar performance profile to the PMN test strategies. CONCLUSIONS: A combination of a nonspecific screening of FVU for PMNs or LET followed by specific testing with EIA with DFA confirmation has superior clinical and cost effective performance for detecting asymptomatic C trachomatis urethritis in young males compared with other strategies. However, an evaluation of the medical, fiscal, and psychological benefits and risks associated with a specific screening strategy for sexually transmitted diseases must be made before adopting a specific strategy for a particular population. PMID- 8411574 TI - A cohort study of thyroid disease in relation to fallout from nuclear weapons testing. AB - OBJECTIVE: To estimate individual radiation doses and current thyroid disease status for a previously identified cohort of 4818 schoolchildren potentially exposed to fallout from detonations of nuclear devices at the Nevada Test Site between 1951 and 1958. DESIGN: Cohort analytic study. SETTING: Communities in southwestern Utah, southeastern Nevada, and southeastern Arizona. PARTICIPANTS: Individuals who were still residing in the three-state area (n = 3122) were reexamined in 1985 and 1986, and information on the subjects' and their mothers' milk and vegetable consumption during the fallout period was obtained by telephone interview (n = 3545). After exclusions to eliminate missing data and confounding factors, 2473 subjects were available for analysis. MAIN OUTCOME MEASURES: Individual radiation doses to the thyroid were estimated by combining consumption data with radionuclide deposition rates provided by the US Department of Energy and a survey of milk producers. Relative risk models adjusted for age, sex, and state were fitted using maximum likelihood to period prevalence data for thyroid carcinomas, neoplasms, and nodules. RESULTS: Doses ranged from 0 mGy to 4600 mGy, and averaged 170 mGy in Utah. There was a statistically significant excess of thyroid neoplasms (benign and malignant; n = 19), with an increase in excess relative risk of 0.7% per milligray. A relative risk for thyroid neoplasms of 3.4 was observed among 169 subjects exposed to doses greater than 400 mGy. Positive but nonsignificant dose-response slopes were found for carcinomas and nodules. CONCLUSIONS: Exposure to Nevada Test Site-generated radioiodines was associated with an excess of thyroid neoplasms. The conclusions are limited by the small number of exposed individuals and the low incidence of thyroid neoplasms. PMID- 8411575 TI - Bacon therapy and furuncular myiasis. AB - OBJECTIVE: To evaluate a simple, noninvasive method for removing fly larvae from patients with furuncular myiasis. DESIGN: Case series. SETTING: Ambulatory office of a tertiary care center. PATIENTS: Three patients who presented with Dermatobia hominis infestation. INTERVENTION: The patients with D hominis infestation were treated with the application of bacon fat over the larval apertures. MAIN OUTCOME MEASURE: Removal of intact larvae. RESULTS: Within 3 hours of the application of bacon, the larvae had migrated sufficiently far out of the skin to be removed with tweezers. Ten larvae were removed with this method. There were no treatment failures or complications. CONCLUSIONS: Furuncular myiasis will be seen more frequently in temperate areas as individuals travel to endemic areas. We describe the clinical characteristics of myiasis and a simple method of treatment that permits rapid diagnosis and cure. PMID- 8411576 TI - Dose-intensive therapy for breast cancer. PMID- 8411577 TI - Users' guides to the medical literature. I. How to get started. The Evidence Based Medicine Working Group. PMID- 8411578 TI - Users' guides to the medical literature. PMID- 8411579 TI - Screening young men for chlamydial infection. PMID- 8411580 TI - A piece of my mind. China tales. PMID- 8411581 TI - Physical restraint use in nursing home patients with dementia. PMID- 8411582 TI - Who keeps the gate? PMID- 8411583 TI - A piece of my mind. A vacation fit for a sobaka (dog). PMID- 8411584 TI - Children's hospitals say about health care reform, 'one size won't fit all'. PMID- 8411585 TI - Nobel prize recognizes 'split gene' research; other pioneers in genetics studies also honored. PMID- 8411586 TI - NIH weighs in for active disease-prevention role. PMID- 8411587 TI - Simpler vaccine schedules target more tykes. PMID- 8411588 TI - From the Agency for Health Care Policy and Research. PMID- 8411589 TI - From the Centers for Disease Control and Prevention. Outbreaks of Salmonella enteritidis gastroenteritis--California, 1993. PMID- 8411590 TI - From the Centers for Disease Control and Prevention. Morbidity surveillance following the midwest flood--Missouri, 1993. PMID- 8411591 TI - Psychotropic medications during pregnancy: risk to the fetus. PMID- 8411592 TI - Psychotropic medications during pregnancy: risk to the fetus. PMID- 8411593 TI - Norms for the mini-mental state examination. PMID- 8411594 TI - Outcomes in elderly patients after intensive care. PMID- 8411595 TI - Outcomes in elderly patients after intensive care. PMID- 8411596 TI - Postmarketing surveillance: beyond MEDWatch. PMID- 8411597 TI - HIV-infected patients participating in autologous blood programs. PMID- 8411598 TI - HIV-infected patients participating in autologous blood programs. PMID- 8411599 TI - Plasma endothelins in sepsis syndrome. PMID- 8411600 TI - Measles vaccination in pediatric emergency departments during a measles outbreak. AB - OBJECTIVE: To determine the proportion of preschool-aged patients attending two inner-city hospital pediatric emergency departments (EDs) who were eligible for measles vaccination, to describe their demographic and clinical characteristics, and to assess the performance of the ED immunization programs that were implemented during a measles outbreak in vaccinating eligible children. DESIGN: Cross-sectional study. SETTING: Pediatric EDs of two urban hospitals in Chicago, Ill, in 1989. PARTICIPANTS: Children 6 months to 5 years of age seen in the EDs. INTERVENTION: None. MAIN OUTCOME MEASURES: The proportion of preschool-aged patients attending the two EDs who were eligible for measles vaccination and the proportion of vaccine-eligible children who were given measles vaccine. RESULTS: Of 508 ED patients at hospital A and 255 patients at hospital B, 18% and 29%, respectively, were considered to be vaccine eligible. The most common discharge diagnoses of eligible patients were viral syndrome, otitis media, and minor trauma. Of vaccine-eligible patients, 59% at hospitals A and B were not vaccinated in the ED. At hospital B, patients with an infectious or respiratory disease diagnosis were less likely to be vaccinated than those with other diagnoses (P < .05). CONCLUSIONS: Many children seen in these EDs were eligible for measles vaccination, and many eligible patients were not vaccinated. During community outbreaks of measles, optimal vaccination programs in pediatric EDs could increase vaccination coverage among inner-city preschool-aged children who may have limited access to health care. PMID- 8411601 TI - Accuracy of immunization histories provided by adults accompanying preschool children to a pediatric emergency department. AB - OBJECTIVE: Because some have advocated the use of emergency departments to administer delayed childhood immunizations, we evaluated the accuracy of immunization histories obtained in this setting by comparison with medical records of inner-city health care facilities. DESIGN: Questionnaires were orally administered to adults accompanying children to the emergency department. Individual medical records were reviewed. SETTING: Pediatric emergency department at Wyler Children's Hospital, University of Chicago and 68 inner-city primary care clinics. PATIENTS: Children aged 3 to 65 months registering for medical care. Of the sample, 98% were African American; 75% were Medicaid recipients. MAIN OUTCOME MEASURES: Adults' knowledge of immunization histories, immunization cards, and medical records compared with American Academy of Pediatrics/Immunization Practices Advisory Committee recommendations. RESULTS: Of the accompanying adults, 64% stated that their child's general immunization status was "up-to-date"; 65% of these had clinic records confirming that status. Only 8% of specific regimens stated by these adults accurately matched those found in clinic records. Moreover, 45% of adults accompanying children at least 16 months and older provided inaccurate information regarding previous receipt of measles immunization. CONCLUSION: Information provided by accompanying adults (from recall or from immunization cards) is inadequate to determine accurately which preschoolers in the pediatric emergency department are delayed in immunizations. PMID- 8411602 TI - A prospective study of lipoprotein(a) and the risk of myocardial infarction. AB - OBJECTIVE: To assess prospectively the risk of myocardial infarction (MI) associated with elevated levels of lipoprotein(a) (Lp[a]). DESIGN: Nested case control study using prospectively collected plasma samples. SETTING: Participants in the Physicians' Health Study. PARTICIPANTS: A total of 14,916 male physicians aged 40 to 84 years with no prior history of MI or stroke provided plasma samples at baseline and were followed up prospectively for an average period of 60.2 months. Samples from 296 physicians who subsequently developed MI were analyzed for Lp(a) level together with paired controls, matched for smoking status and age. MAIN OUTCOME MEASURE: Fatal or nonfatal acute MI. RESULTS: The distribution of Lp(a) level among cases was virtually identical to that of controls (P = .88), and there was no significant difference between groups for median Lp(a) levels (103.0 mg/L vs 102.5 mg/L; P = .73). In analyses controlling for age and smoking status, we found no evidence of association between increasing level of Lp(a) and risk of MI (relative risks from lowest to highest quintiles of Lp(a): 1.00, 0.97, 0.83, 0.88, and 1.07; P for trend = .93) or a threshold effect at any prespecified cutoff of Lp(a) level (relative risks associated with Lp[a] levels above the 25th, 50th, 75th, 90th, and 95th percentiles of the control distribution, respectively: 1.04, 1.00, 1.19, 1.00, and 1.07; all P values nonsignificant). Further adjustment for both lipid and nonlipid cardiovascular risk factors had no material impact. CONCLUSIONS: In this prospective study of predominantly middle-aged white men, we found no evidence of association between Lp(a) level and risk of future MI. These data do not support the use of Lp(a) level as a screening tool to define cardiovascular risk among this population. PMID- 8411603 TI - Classic nephropathic cystinosis as an adult disease. AB - OBJECTIVE: To delineate the clinical characteristics of infantile nephropathic cystinosis in adult patients who have undergone renal transplantation. DESIGN: Case series. SETTING: Clinical research unit. PATIENTS: All 36 adult patients with nephropathic cystinosis referred to the National Institutes of Health. OUTCOME MEASURES: Longevity, growth, renal allograft survival, visual acuity, endocrine insufficiency, myopathy and swallowing dysfunction, cerebral calcifications, and occupational status. RESULTS: Of the 36 patients, seven were dead, five with functioning allografts. The 1-year and 5-year graft survival rates for 30 cadaveric allografts were 90% and 75%, respectively. The patients' mean height and weight were severely retarded. Five patients were legally blind, and three others had severely impaired vision in one eye. Thirty-one (86%) of 36 patients required thyroid hormone replacement therapy. One third had a distal myopathy, and 21 had moderate to severe swallowing abnormalities. Eight patients had cerebral calcifications on computed tomographic scan. Despite these complications, the sighted patients engaged in a normal variety of occupations. Only 11 patients were receiving adequate cystine-depleting therapy with cysteamine (mercaptamine) or phosphocysteamine. CONCLUSIONS: Adult patients with nephropathic cystinosis suffer serious complications of the disease. PMID- 8411604 TI - Pleurodynia among football players at a high school. An outbreak associated with coxsackievirus B1. AB - OBJECTIVE: Enteroviral outbreaks involving athletic teams have been described, although the mode of transmission has been unclear. In September 1991, an outbreak of pleurodynia among high school football players provided an opportunity to identify possible modes of transmission. DESIGN: Retrospective cohort outbreak investigation. SETTING: Public high school in upstate New York. RESULTS: Illness was reported by 17 (20%) of the football players. Behaviors involving contact with common water containers were associated with illness, including eating ice cubes from the team ice chest (relative risk [RR], 9.2; 95% confidence interval [CI], 1.3 to 65.5) and drinking water from the team cooler (RR, 6.3; 95% CI, 1.5 to 25.7). Coxsackievirus B1 was isolated in four (50%) of the eight stool specimens collected. CONCLUSIONS: Contamination of common water containers by an infected player may have contributed to or initiated the outbreak. In addition to discouraging direct oral contact with common drinking containers, use of individual water containers and ice packs for injuries was recommended. PMID- 8411605 TI - Actual causes of death in the United States. AB - OBJECTIVE: To identify and quantify the major external (nongenetic) factors that contribute to death in the United States. DATA SOURCES: Articles published between 1977 and 1993 were identified through MEDLINE searches, reference citations, and expert consultation. Government reports and complications of vital statistics and surveillance data were also obtained. STUDY SELECTION: Sources selected were those that were often cited and those that indicated a quantitative assessment of the relative contributions of various factors to mortality and morbidity. DATA EXTRACTION: Data used were those for which specific methodological assumptions were stated. A table quantifying the contributions of leading factors was constructed using actual counts, generally accepted estimates, and calculated estimates that were developed by summing various individual estimates and correcting to avoid double counting. For the factors of greatest complexity and uncertainty (diet and activity patterns and toxic agents), a conservative approach was taken by choosing the lower boundaries of the various estimates. DATA SYNTHESIS: The most prominent contributors to mortality in the United States in 1990 were tobacco (an estimated 400,000 deaths), diet and activity patterns (300,000), alcohol (100,000), microbial agents (90,000), toxic agents (60,000), firearms (35,000), sexual behavior (30,000), motor vehicles (25,000), and illicit use of drugs (20,000). Socioeconomic status and access to medical care are also important contributors, but difficult to quantify independent of the other factors cited. Because the studies reviewed used different approaches to derive estimates, the stated numbers should be viewed as first approximations. CONCLUSIONS: Approximately half of all deaths that occurred in 1990 could be attributed to the factors identified. Although no attempt was made to further quantify the impact of these factors on morbidity and quality of life, the public health burden they impose is considerable and offers guidance for shaping health policy priorities. PMID- 8411606 TI - Do-not-resuscitate orders in intensive care units. Current practices and recent changes. AB - OBJECTIVES: To describe the characteristics of patients with do-not-resuscitate (DNR) orders and the frequency and timing of these orders in a representative sample of intensive care units (ICUs) and to compare practices from 1980 to 1990. DESIGN: Prospective inception cohort. SETTING: A total of 42 ICUs in 40 US hospitals with 200 or more beds; 26 randomly selected hospitals and 14 large, tertiary care hospitals that volunteered to be studied. PARTICIPANTS: A consecutive sample of 17,440 ICU admissions from 1988 to 1990. MEASUREMENTS: Patient demographic characteristics, comorbid conditions, disease, and physiological abnormalities. MAIN OUTCOME MEASURES: Frequency and timing of DNR orders; ICU resource use before and after DNR orders; and patients' hospital and ICU discharge status. RESULTS: Physicians wrote DNR orders for 1577 ICU admissions (9%) (hospital range, 1.5% to 22%). Patients with ICU DNR orders were older, more functionally impaired, had more comorbid illness, a higher severity of illness, and required the use of more ICU resources compared with patients without DNR orders. Compared with data from a similar survey from 1979 to 1982, ICU DNR orders were more frequent in 1988 to 1990 (9% vs 5.4%; P < .001) and preceded 60% of all in-unit deaths compared with only 39% in 1979 to 1982 (P < .001). Do-not-resuscitate orders were written sooner (for 3.6% vs 2.0% of patients on day 1 in the ICU) and patients with DNR orders remained in the ICU longer in 1988 to 1990 (2.8 vs 1.4 days) than in 1979 to 1982, and had lower ICU and hospital mortality rates (64% vs 74%, P < .001; and 85% vs 94%, P < .001). CONCLUSIONS: Over the last decade physicians and patients' families set limits earlier and more frequently in cases likely to have poor outcomes. We attribute this change to a greater dialogue about setting limits to care and a greater knowledge of treatment outcomes among physicians and families. These changes in practice preceded implementation of the Patient Self-determination Act, designed to ensure patient autonomy for decisions about life-sustaining therapy. PMID- 8411607 TI - The rational clinical examination. Does this patient have splenomegaly? PMID- 8411608 TI - Vaccination in pediatric emergency departments. PMID- 8411609 TI - Has lipoprotein 'little' (a) shrunk? PMID- 8411610 TI - Omeprazole: is it the answer of peptic ulcer disease? PMID- 8411611 TI - Omeprazole vs ranitidine in the healing of duodenal ulcer. AB - To assess the comparative efficacy of omeprazole 20 mg in the morning versus ranitidine 150 mg twice a day, in the healing of duodenal ulcer, 43 endoscopically verified cases were randomly allocated to 2 or 4 weeks (if ulcer did not heal at 2 weeks) treatment. Two cases were excluded due to deviation from the protocol. Of 21 cases treated with omeprazole 15 (71%) healed at two weeks and all (100%) at 4 weeks. Similarly of 20 cases treated with ranitidine, 14 (70%) healed at 2 and 18 (90%) at 4 weeks (P = NS). Two patients who did not heal with ranitidine, when crossed over to omeprazole healed at 4 weeks. No significant difference in the rate of pain relief, clinical or laboratory parameters or adverse effects were noted in either group. Both the drugs were well tolerated and were potent healer of duodenal ulcer; but omeprazole appears to be superior in the healing of resistent ulcers. PMID- 8411612 TI - Suggested reference ranges in clinical chemistry for apparently healthy males and females of Pakistan. AB - Seven hundred and eighty six apparently healthy males (418) and females (368) aged 0-69 years were randomly selected for estimation of reference ranges of 24 serum analytes at the clinical chemistry laboratory of The Aga Khan University Hospital (AKUH). Of the total study samples, 56% (439/786) were in the paediatric age group (0-14 years) and 44% (347/786) in the adult (15 > or = 60 years) group. Beckman Astra Ideal Autoanalyzer was used for all the estimations. Mean and standard deviations (SD) were calculated for each of the age groups. Reference ranges were calculated following standard methods of the International Federation of Clinical Chemistry (IFCC) and International Committee for Standardization in Haematology (ICSH). PMID- 8411613 TI - The safety of epidural analgesia in labour and its effect on delivery--a case control study in Pakistani women. AB - To assess the safety and effect of epidural analgesia on the course of labour and delivery in Pakistani women, a retrospective case control study was conducted from November, 1986 to November, 1991 (5 years) at the Aga Khan University Medical Centre, Karachi. All patients (n = 64) who received epidural analgesia for labour (cases) were compared with randomly selected patients (n = 18) who did not receive epidural analgesia during labour (controls). The cases and controls were matched for age, height, body mass index, parity, use of oxytocin, presentation and weight of the foetus. There was no significant difference (P > 0.05) between the two groups in duration of labour, caesarean section rate and foetal apgar scores at 1 and 5 minutes after birth. The incidence of malposition of foetal vertex at delivery and that of instrumental (forceps) deliveries was significantly higher (P < 0.05 and < 0.01 respectively) in the epidural group as compared to controls. The incidence of complications was low and the acceptance and tolerance of epidural analgesia was good in our patients. PMID- 8411614 TI - Nutritional aspects of mammary carcinogenesis: a case-control study. AB - Dietary factors are believed to play an important role in mammary carcinogenesis. International correlations, case-control and cohort studies have associated the incidence and mortality from breast cancer with high fat consumption in the form of meat, gravy and dairy products. Most of these studies have been conducted in the developed countries. Due to paucity of data from developing countries, we conducted a case-control study to evaluate the role of nutritional factors in mammary carcinogenesis. This prospective study was conducted in the oncology clinic at a university hospital. On a detailed questionnaire, information was collected from 80 patients with histologically proven breast cancer (cases) and 80 normal healthy subjects (controls). Information was collected on several patients characteristics as well as intake of 44 different food items. Special care was taken to exclude any recent changes in diet, induced in cases since learning the diagnosis of breast cancer. Interviews were conducted by the same interviewer and lasted approximately 20 minutes. Analysis of data reveal no significant difference in patients characteristics between cases and controls. Similarly, no significant difference was found in the intake of most dietary items including meat and dairy products. However, a significant difference was observed in the consumption of fish, vegetable and fruits (P = 0.05). We conclude that our study fails to support the fat-breast cancer hypothesis. It also suggests a potential role of other dietary items such as fish, vegetables and fruits in mammary carcinogenesis. PMID- 8411615 TI - Desirability of using buprenorphine and diazepam as an adjunct to atropine in patients undergoing fibreoptic bronchoscopy. AB - This study endeavours to evaluate the need for sedation in fibreoptic bronchoscopy and response of the patients when sedation is not used. One hundred and five patients divided into three groups of 35 patients each, received buprenorphine (0.3 mg), diazepam (10 mg) and normal saline (0.5 ml) respectively along with atropine (0.6 mg) in each case. The study emphasizes that sympathetic explanation of the details of the procedure to the patients, administration of careful topical anaesthesia and avoidance of sedation makes the procedure acceptable, safe, comfortable with less complications and the need for admitting the patients to the ward is by and large avoided. PMID- 8411616 TI - Tuberculous mastitis mimicking malignancy--a case report with review of literature. PMID- 8411617 TI - Aggressive angiomyxoma of vulva. PMID- 8411618 TI - Caffeic acid. PMID- 8411619 TI - d-Limonene. PMID- 8411620 TI - IQ (2-amino-3-methylimidazo[4,5-f]quinoline). PMID- 8411621 TI - MeIQ (2-amino-3,4-dimethylimidazo[4,5-f]quinoline). PMID- 8411622 TI - MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline). PMID- 8411623 TI - PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine). PMID- 8411624 TI - Aflatoxins. PMID- 8411625 TI - Toxins derived from Fusarium graminearum, F. culmorum and F. crookwellense: zearalenone, deoxynivalenol, nivalenol and fusarenone X. PMID- 8411626 TI - Salted fish. PMID- 8411627 TI - Toxins derived from Fusarium moniliforme: fumonisins B1 and B2 and fusarin C. PMID- 8411628 TI - Toxins derived from Fusarium sporotrichioides: T-2 toxin. PMID- 8411629 TI - Ochratoxin A. PMID- 8411630 TI - Pickled vegetables. PMID- 8411631 TI - Safety and accuracy of dipyridamole thallium myocardial scintigraphy in elderly patients. AB - This study examined mainly the adverse effects of 201Tl myocardial scintigraphy with dipyridamole (D-Tl) in 73 elderly patients over 70 years old in comparison with those in 65 younger patients. Fifty-five of 73 elderly patients (75%) and 49 of 65 younger patients (75%) had a persistent or dipyridamole-induced perfusion defect on D-Tl. The hemodynamic changes induced by dipyridamole as well as the incidence of cardiac and noncardiac adverse effects were similar in both groups and no serious adverse effect occurred in either group. Secondly, we examined the procedure's usefulness for detecting ischemic heart disease in elderly and younger patients. Dipyridamole induced perfusion defect was noted in 21 elderly patients and in 24 younger patients (N.S.). Among the patients in whom coronary angiography was performed, significant coronary artery stenosis was found in 5 of 8 elderly patients and 17 of 20 young patients (N.S.). In patients with one or two-vessel disease, the area with dipyridamole induced ischemia was concordant with the stenotic area seen on coronary angiography in 3 of 3 elderly patients and 12 of 13 younger patients (N.S.). Thus, the safety and usefulness of D-Tl for detecting myocardial ischemia were comparable in elderly and young patients. PMID- 8411632 TI - Pleomorphic ventricular tachycardia arising from a new site during antiarrhythmic drug therapy. AB - We analyzed the site of VT origin and the induction mode of VT in 9 patients who showed new VT morphologies with different bundle branch block patterns after administering antiarrhythmic drug(s). In all patients, VT exhibiting the clinical morphology was induced in the drug free state. (1) VT origin: In 6 patients, VTs showing LBBB pattern had a site of origin at the right ventricular free wall, and VTs with RBBB pattern originated from the left ventricular free wall. VT from the intraventricular septum of the right ventricle showed RBBB pattern in two patients and VT with LBBB pattern arose from the posteroseptum of the left ventricle in one patient. (2) VTs with new morphologies: After administering drug(s), VTs with new morphologies were induced in 18 studies and the mean cycle length of these VTs was not different from that in the control study. Among them, the induction mode was less aggressive in 4 of 7 drug studies and more aggressive in 1 study. (3) VTs with the same morphology: VTs with morphologies identical to those of the clinical VTs were induced in 15 studies. However, the drugs' effect was evident. The mean cycle length of these VTs was significantly prolonged, and VTs were induced by less aggressive modes or at longer coupling intervals. IN CONCLUSION: (1) After administering drug(s), different electrophysiologic characteristics were observed between the VTs with new morphologies and the VTs with the same morphology. (2) If a new VT was induced by less aggressive modes after administering drug(s), the drug(s) might act to facilitate inducibility: proarrhythmic effect. PMID- 8411633 TI - Pronounced ST-segment depression during paroxysmal supraventricular tachycardia. AB - To determine the clinical significance of ST-segment depression observed in paroxysmal supraventricular tachycardia (PSVT), we evaluated the 12-lead electrocardiogram (ECG) during spontaneous PSVT in 54 patients (27 men and 27 women: mean age +/- SD; 47 +/- 18 years), who came to our clinic for the treatment of PSVT. Coronary angiography was performed in 16 patients (16 to 74 years; mean = 50 +/- 18) and treadmill exercise testing was performed in 21 patients. A cardiac electrophysiological study was carried out in 24 patients. During PSVT, ST-segment score was calculated as the sum of the ST-segment depression in 12 leads. The correlations between the ST-segment score, PSVT rate and age of the patient were analyzed as follows: The most significant positive correlation was observed between the ST-segment score and the PSVT rate (r = 0.615, p < 0.000001). The next most significant correlation was found between the PSVT rate and the age of the patient (r = -0.500, p = 0.00011). A negative correlation was also observed between the ST-segment score and the age of the patient (r = -0.429, p = 0.0012). In 13 of 16 patients, coronary angiography did not reveal any significant (> or = 75% in area) stenosis. Exercise testing induced significant ST-segment depression in 3 patients, of whom two had significant coronary artery lesions. PSVT was due to atrioventricular reentry via an overt (n = 3) or concealed accessory pathway (n = 15), atrioventricular nodal reentry (n = 5) and sinus node reentry (n = 1). In conclusion, patients with a faster PSVT rate revealed more pronounced ST-segment depression than did those with a slower PSVT rate, possibly reflecting the modified repolarization process instead of coronary artery involvement. PMID- 8411634 TI - The relationship between ambulatory blood pressure and physical activity in young and older shiftworkers. A quantitative assessment of physical activity using a microcomputer with acceleration sensor. AB - We studied the relationship between physical activity and ambulatory blood pressure (BP) in young and older shiftworkers by simultaneous recordings of activity, blood pressure and pulse rate (PR). Activity was assessed using Activetracer, a self-contained microcomputer with an acceleration sensor, attached to a waist belt. Ambulatory BP was monitored every 30 minutes for 48 hours with a TM2421. Three types of hemodynamic responses were noted in relation to the physical activity. The balance type, in which both BP and PR increase with physical activity, was observed in 5 of 10 young cases (50%) but only in 1 of 7 older cases (14.3%). The BP response type, in which the BP increases with no change in the PR, was observed in 6 of the 7 older (86%) but only in 3 cases in the young group (30%). The PR response type, in which only the PR increase correlated with activity, was observed in 2 cases in the young group (20%) and none in the older group. The difference in systolic BP between periods of activity and rest in the older shiftworker was significantly larger than that in the young group (15.9 +/- 6.4 vs. 5.9 +/- 6.6 mmHg, p < .01), although no significant difference was observed in diastolic BP. In contrast, the increase in pulse rate after movement was significantly higher in the young group (4.4 +/- 4.0 vs. 9.0 +/- 4.8 bpm, p < .05). Thus, the fluctuation of the systolic BP was more dependent on physical activity in the older group, whereas PR variations correlated with the physical activity in the young group. PMID- 8411635 TI - Calcium concentration during anoxic perfusion modifies post-ischemic hypercontraction in the canine heart. AB - The purpose of this study was to clarify the role of calcium flux in the pathogenesis of transient overshoot in regional myocardial contractile function after brief ischemia (post-ischemic hypercontraction). Six open-chest anesthetized dogs were examined. The left anterior descending coronary artery (LAD) was cannulated with a bypass system from the left carotid artery. Two minutes of total coronary occlusion of the LAD resulted in a post-ischemic hypercontraction 1 minute after reperfusion. Post-ischemic hypercontraction was abolished after reperfusion following 2 minutes of perfusion with anoxic Krebs Henseleit solution containing 2.5 mM calcium. Post-ischemic hypercontraction occurred after calcium-free anoxic perfusion. The regional myocardial contractile function remained depressed 1 minute after reperfusion following anoxic perfusion with 5.0 mM calcium solution. Thus, post-ischemic hypercontraction was modified by the calcium concentration during anoxia. Alteration in transsarcolemmal calcium influx during reperfusion, which was modified by alterations in the calcium environment during anoxia, could be responsible for this phenomenon. PMID- 8411636 TI - Effects of pilsicainide on the atrial fibrillation threshold in guinea pig atria. A comparative study with disopyramide, lidocaine and flecainide. AB - The effects of pilsicainide, a new class Ic antiarrhythmic agent, on the atrial fibrillation threshold (AFT), the atrial effective refractory period (ERP), and the interatrial conduction time (ACT) in Langendorff-perfused guinea pig hearts were investigated. These effects were compared with those of disopyramide, lidocaine and flecainide. Whole guinea pig heart was perfused with Tyrode's solution containing acetylcholine (3 x 10(-7) M). Three indices were measured before and after the administration of the test drugs using right atrial extrastimulus and high frequency stimulation. Pilsicainide, disopyramide and flecainide (> or = 10(-6) M) all significantly increased the AFT. Both pilsicainide and flecainide (> or = 3 x 10(-6) M) significantly prolonged the ERP, but this prolongation was less pronounced than that observed with disopyramide. The ACT was significantly prolonged with pilsicainide (> or = 10( 6) M), and this prolongation was greater than that observed with disopyramide but less than that with flecainide. Lidocaine had no effects on any of the indices measured. In conclusion, pilsicainide had a preventive effect on the atrial fibrillation induced by a combination of acetylcholine and high frequency stimulation in guinea pig hearts, by increasing the atrial ERP and slowing the interatrial conduction. These effects may explain, in part, the clinical effectiveness of this drug on paroxysmal atrial fibrillation. PMID- 8411637 TI - In vivo profile of myocardial energy metabolism of pressure-overloaded rat. AB - Cardiac energy metabolism of pressure-overloaded rat hearts was examined under in vivo and in vitro conditions. Two, 4 and 6 weeks after constriction of the abdominal artery, the hemodynamic and metabolic profiles of hearts in vivo and of perfused hearts were determined. Significant increases in left ventricular weight/body weight (30 to 45% increase relative to the sham group), systolic and diastolic blood pressure (22 to 33% increase) and pressure-rate product (31 to 33% increase) were observed 2, 4 and 6 weeks after the operation, and a slight but significant decrease in heart rate was observed at 2 weeks after the operation. Tissue hydroxyproline content increased (17 to 93%) with time after pressure-overload. These findings are indicators of pressure-overloaded cardiac hypertrophy. The total high-energy phosphates of the in vivo rat myocardium under artificial respiration were lower than those of sham-operated rat myocardium 2 (23%) and 4 weeks (21%), but not 6 weeks after aortic constriction. The maximal oxygen consumption rates of mitochondria, when determined in the skinned cardiac fibers, also decreased 2 (47%) and 4 weeks (36%), but reversed 6 weeks after pressure-overload. However, the myocardial ATP, a utilizing form of high-energy phosphate, of pressure-overloaded rat myocardium remained normal at all times after cardiac hypertrophy. This suggests that alterations in hemodynamic variables of in vivo pressure-overloaded rats may not be attributable to a reduction in the myocardial energy production. In the perfused hearts isolated from pressure-overloaded rats, tissue ATP levels were similar to those of sham operated rats, although the tissue creatine phosphate tended to be reduced in the pressure-overloaded animals at all stages of cardiac hypertrophy examined. Only a marginal decrease in the tissue high-energy phosphate (13%) was observed 4 weeks after the operation relative to that of sham-operated rats. In contrast, the developed tension of the perfused pressure-overloaded rat hearts was consistently lower (27 to 36%) than that of the sham-operated rat hearts. The results suggest that the high-energy phosphate levels of pressure-overloaded rat myocardium in vitro are unlikely to account for the observed decline in cardiac contractile function. The reduction of myocardial high-energy phosphates of pressure overloaded rats may be due to an adaptative change rather than a causal events. PMID- 8411638 TI - Clonidine improves central attenuation of the baroreflex in spontaneously hypertensive rats. AB - To determine whether clonidine can improve the central alteration of the baroreflex in SHR, the centrally cut end of the aortic depressor nerve was electrically stimulated in SHR with intravenously administered clonidine. Aortic depressor nerve (ADN) stimulation elicited depressor and sympatho-inhibitory responses in a frequency-dependent manner in WKY and SHR. These responses were significantly smaller in SHR than in WKY. The attenuated depressor and sympatho inhibitory responses to ADN stimulation in SHR were restored following i.v. injections of clonidine, although the drug did not affect the responses to ADN stimulations in WKY. These findings suggest that clonidine can improve central attenuation of the baroreflex in hypertension. PMID- 8411639 TI - Loss of vasoreactivity by laser thermal energy or argon laser irradiation. AB - Vasoreactivity of laser-treated vessels was investigated in two different experimental conditions. The canine left circumflex coronary artery (LCx) was lased under perfusion with Krebs-bicarbonate buffer by means of a thermal laser (hot-tip probe, HT) at 7 W for 6 seconds and an argon laser beam through a 300 microns optical fiber at 3 W (tip power) for 1 second at 12 spots. A nontreated segment of the LCx served as a control. Two 3-mm long segments were obtained from the treated segment: one to measure the results of potassium (K) induced contraction, and another 3, 4 diaminopyridine (DAP; K channel inhibitor) induced contraction. In 11 instances, coronary angiography of the perfused artery showed less than 50% stenosis after laser treatment. The segments were then mounted isometrically with 1 g tension in Krebs-bicarbonate buffer. Contraction was induced either with 30 mM KCI or 10(-2) M DAP and expressed as developed tension (gram; g). KCI induced vasocontraction of 4.15 +/- 0.93 g in the control, 0.33 +/ 0.71 g in laser irradiated segments (P < 0.0001 vs control), and 0.02 +/- 0.06 g in thermally-treated segments (P < 0.0001 vs control). DAP induced vasocontraction of 5.21 +/- 1.32 g in the control, 0.39 +/- 0.83 g in laser irradiated segments (P < 0.0001 vs control), and 0.07 +/- 0.13 g in thermally treated segments (P < 0.001 vs control). In 4 instances, more than 50% stenosis remained and additional balloon dilatation reduced the stenosis to less than 50%. The lesions also showed reduced vasoreactivity. In vivo thermal angioplasty resulted in reduced vasoreactivity compared to control in 4 anesthetized dogs. Thus, laser and thermal angioplasty reduced vasoreactivity induced by either KCI or 3, 4 DAP. Neither acetylcholine at 10(-6) M nor papaverine at 10(-4) M was able to induce relaxation of treated segments. In conclusion, 1) the lased coronary artery loses its vasoreactivity to either a constrictive or relaxing agent, 2) although stenosis may be produced by laser energy, additional balloon dilatation can reduce residual stenosis, and 3) laser thermal or argon laser angioplasty may prevent severe coronary spasm. PMID- 8411640 TI - Left ventricular dysfunction due to hypocalcemia in a neonate. AB - Hypocalcemia is a relatively uncommon but reversible cause of left ventricular dysfunction in infants and children. A 30-day-old boy with idiopathic hypocalcemia presented with congestive heart failure and convulsive seizures. He had no evidence of underlying cardiac disease. The cardiac failure responded to calcium therapy. It is suggested that hypocalcemia should be considered as a possible cause of left ventricular dysfunction in infants. PMID- 8411641 TI - Tricuspid regurgitation due to blunt chest trauma. Report of a case and review of the literature. AB - A 25-year-old female developed high-grade atrioventricular block and markedly elevated central venous pressure after sustaining a crushing injury to the chest while driving a car. An echocardiographic examination with color Doppler revealed severe tricuspid regurgitation due to a torn papillary muscle. An extensive review of the literature showed the following: 1) correct diagnosis is often delayed because of coexisting multisystem involvement and the subtleness of abnormal physical signs, 2) identification of abnormally elevated right atrial pressure with a prominent "v" wave, and characteristic electrocardiogram appeared to be the key to early diagnosis, and 3) the final diagnosis may be confirmed by echocardiography with Doppler and/or cardiac catheterization. The role of echocardiographic examination with color Doppler technique deserves special emphasis because the final diagnosis can be easily reached during the acute phase at the bedside noninvasively. PMID- 8411642 TI - Chronic aortic dissection complicated by recurrent obstructive pneumonia. AB - A 68-year-old man with aortic aneurysm who had repeated episodes of obstructive bronchopneumonia is reported. Serial chest X-rays revealed infiltrative shadows in the left lower lung field. A thoracic computed tomogram demonstrated a dissecting descending aorta compressing the left lower bronchi and abnormal shadows, probably inflammatory, distal to the obstructions. Because of signs of impending rupture of the dissected aorta, surgical repair was performed and there has been no recurrence of respiratory infection since. PMID- 8411643 TI - Coronary artery spasm in survivors of cardiac arrest. PMID- 8411644 TI - [Clinical usefulness of measurement of erythropoietin in blood]. AB - Blood erythropoietin (EPO) concentration was measured by radioimmunoassay in 513 patients with various diseases. Untreated polycythemia vera showed lower EPO concentration than normal. Aplastic anemia (AA) revealed the highest EPO level among all anemic diseases in relation to hematocrit value. EPO level of AA patients who underwent bone marrow transplantation was as low as normal subjects even when the anemia has not fully recovered. Paroxysmal nocturnal hemoglobinuria (PNH) showed unusually high EPO concentration among hemolytic anemias. In normal subjects, blood EPO concentration showed a diurnal rhythm that was higher at night than during the daytime. These findings suggest the diagnostic usefulness of measurement of EPO in blood diseases. PMID- 8411645 TI - [Detection of minimal residual clone after sex-mismatched bone marrow transplantation by fluorescent in situ hybridization]. AB - In our previous paper we reported that fluorescent in situ hybridization (FISH) using DYZ1 and DXZ1 was a highly reproducible technique and cellular chimerism consisting of cells of male and female origins could be detected in the order of 0.1%. In the present study FISH was applied to detect minimal residual clones (MRC) in 10 patients with a variety of leukemias who received sex-mismatched bone marrow transplantation (BMT). In 9 patients who had no signs of recurrence, serial FISHs performed on each patient revealed the presence of 0.1 to 3.2% recipient residual clone. On the other hand, in the remaining one patient 10.6% and 16.9% of MRC were found by FISH performed 6 and 10 months after BMT. Immediately thereafter, this patient was diagnosed as in relapse by bone marrow examination. PMID- 8411646 TI - [Phase II clinical study of recombinant human erythropoietin on the anemia associated with multiple myeloma]. AB - Safety and efficacy of recombinant human erythropoietin (epoetin alpha) were investigated in anemic patients with multiple myeloma whose hemoglobin (Hb) concentration was less than 10g/dl. Epoetin alpha (3,000IU/body) was given subcutaneously daily for two weeks and the dosage was increased to 6,000IU, 12,000IU and 24,000IU every two weeks when the increment of Hb was insufficient. Cases in which Hb concentration increased by more than 1g/dl or in which blood transfusion requirements decreased by more than 50% were judged to be effective. The overall rate of efficacy was 52.6% (10/19). Response to epoetin alpha treatment was better in patients whose blood erythropoietin level was relatively low. The majority of patients responded to the treatment with up to 6,000IU/body/day but a dosage of more than 12,000IU/body/day was required in some cases. No serious adverse effects or abnormal laboratory findings were observed. These results suggest that high-dose subcutaneous epoetin alpha treatment is effective for anemia associated with multiple myeloma in terms of increasing Hb concentration and reducing blood transfusion. PMID- 8411647 TI - [Phase II clinical study of recombinant human erythropoietin on the anemia of myelodysplastic syndrome]. AB - The safety and efficacy of recombinant human erythropoietin (epoetin alpha) were investigated in anemic patients with myelodysplastic syndrome (MDS) whose hemoglobin (Hb) concentration was less than 10g/dl. Epoetin alpha was given subcutaneously daily at a dose of 3,000IU/body for two weeks and the dosage was increased to 6,000IU, 12,000IU, and 24,000IU at two week interval when the increment of Hb was insufficient. Patients in whom the Hb concentration increased by more than 1g/dl or whose blood transfusion requirement reduced to below 50% were considered to be cases of effective treatment. The overall rate of effectiveness was 20.6% (7/34). Response to epoetin alpha treatment was better in patients with refractory anemia (RA) or RA with ringed sideroblasts (RARS). The high dose epoetin alpha (12,000-24,000IU/body/day) was required for the patients to respond. These results suggest that the high dose subcutaneous epoetin alpha treatment is effective for the anemia associated with MDS in terms of increasing Hb concentration and reducing blood transfusion. PMID- 8411648 TI - [Involvement of the intraabdominal lymph nodes in a case of subacute necrotizing lymphadenitis]. AB - A 40-year-old woman was admitted to our hospital because of fever, polyposia and polyuria in October 1990. The fasting blood sugar level was 471mg/dl and abdominal CT showed enlargement of the liver, kidneys, and intraabdominal lymph nodes. Although severe diabetes mellitus was controlled with insulin, intraabdominal lymph node swelling continued. Lymph node biopsy was performed under laparotomy. Four from intraabdominal lymph nodes, in addition to one from the left axillary lymph nodes. Four showed findings of non specific lymphadenitis, but one specimen obtained from near the right kidney demonstrated focal necrosis and invasion of macrophages and immunoblasts, which were compatible with the features of subacute necrotizing lymphadenitis (SANL). In SANL, lymphadenopathy is usually observed in the cervical region of young female and the involvement of intraabdominal lymph nodes is quite rare. This patient is the second case of SANL involving an intraabdominal lymph node reported in Japan, and it is suggested that SANL should be considered as a causative disorder of intraabdominal lymphadenopathy. PMID- 8411649 TI - [Long term follow-up of three cases of acute myeloid leukemia associated with inv(16)(p13q22)]. AB - Three cases of acute myeloid leukemia associated with inv(16(p13q22) were followed up for over 5 years. This chromosome aberration is generally thought to be a good prognostic factor. However, it is also reported that these patients are apt to relapse and have relatively high frequency of central nervous system (CNS) involvement. The first patient (M4Eo), who died of gastric cancer about 5 years after the initial treatment without frank relapse, did not have prophylactic therapy for CNS involvement. The second patient (M5b) developed meningeal leukemia and myeloblastoma of the brain, showing similar findings on CT scan to cases reported by Holms et al. He was treated successfully with whole brain irradiation and intrathecal injection of ara-C and MTX, and intracranial tumor disappeared on CT and MR imaging. He has been enjoying a good quality of life without any complication for over ten years after the initial diagnosis. The third patient (M4Eo) relapsed once but reentered complete remission with relative ease and we used an intrathecal injection prophylactically. This case has been followed up as an outpatient for more than 5 years since onset. On the basis of these findings, it may be concluded that these leukemia patients with inv(16)(p13q22) have good prognosis and can be cured with chemotherapy. PMID- 8411650 TI - [Rapid development of primary amyloidosis after two years' follow-up of monoclonal gammopathy of undetermined significance (MGUS)]. AB - Recently, we have encountered an unusual case of "monoclonal gammopathy of undetermined significance (MGUS)" in which some clinical features indicative of primary and amyloidosis (PA) were observed two years after the diagnosis of MGUS. The patient, born in 1925, was diagnosed of having MGUS (IgG-lambda type M component level; 1 g/dl) in June 1990. The following clinical features occurred in close succession within five months after a stable course of two years: general malaise, abdominal, distension, pretibial edema and facial puffiness, ecchymoses on the chest wall, and dyspnea on effort. The biopsy specimens from the skin and gastric mucosa revealed amyloid deposition. The M-component levels in the serum and urine as well as the number of bone marrow plasma cells remained unchanged. The various kinds of laboratory examinations indicated that systemic amyloidosis rapidly developed within five months. It seems quite likely that the heart, liver, and spleen may be affected with amyloidosis. It is noteworthy that PA can occur without an increase in the serum M-component level at any time after the diagnosis of MGUS. An early diagnosis of PA as well as multiple myeloma should be kept in mind in the follow-up study of patients with MGUS. PMID- 8411651 TI - [Neutrophilic myelofibrosis; a case report]. AB - A 53-year-old male was admitted to our hospital with abdominal pain. Physical examination revealed marked splenomegaly. The white blood cell count increased to 5.8 x 10(4)/microliters. Bone marrow biopsy showed hypercellularity with a moderate increase in reticulin fiber. Chromosomal analysis showed 47, XY, +9q-, 9q- without Ph1 chromosome and bcr-abl rearrangement. MCNU therapy was successful in reducing the white blood cell count and splenomegaly. It is likely that the diagnosis of our patient is compatible with the neutrophilic myelofibrosis described by Stewart, et al. PMID- 8411652 TI - [Multiple myeloma presenting with amyloid arthropathy]. AB - Amyloid arthropathy rarely occurs in patients with multiple myeloma (MM) or primary amyloidosis (PA). Amyloid infiltration in and about the joints may be so extensive as to simulate the findings of rheumatoid arthritis. Some cases have been reported in which the articular manifestations were present for many months prior to the diagnosis of amyloid arthropathy. The delay of the diagnosis can result in the development of a fatal complication of MM or PA, which is not always unavoidable. We have encountered an unusual case of MM which the articular manifestations were present prior to the diagnosis of MM. A 59-year-old woman had a four-month history of hypesthesia in the median-nerve distribution of both hands and polyarthralgia. Far advanced renal insufficiency was evident, but its etiology was not determined. The patient was maintained on hemodialysis. The shoulders, wrists, hips and finger joints were symmetrically involved with articular swelling. All of these joints showed the avid uptake of Tc-99m (V) DMSA. The serum and urine immunoelectrophoresis demonstrated the presence of IgG lambda type M-component and lambda type Bence Jones proteins, respectively. The bone marrow findings and bone roentgenograms supported the diagnosis of MM. Biopsy specimens from the synovial membrane revealed amyloid deposition. Her condition was much improved with melphalan and prednisolone. PMID- 8411653 TI - [Clonal change of HTLV-1 infected lymphocytes before onset of adult T-cell leukemia/lymphoma]. AB - A 73-year-old HTLV-1 infected male developed overt ATL following 3-years observation, and the clonality of HTLV-1 infected cells changed before overt ATL onset. 10 micrograms of DNA, extracted from mononuclear cells was digested with PstI, and the clonal proliferation of HTLV-1 infected cells was examined by Southern blotting with LTR probe. 4 bands were observed 3 years prior to ATL onset, and 2 different bands were detected 2 years and 6 months later. This suggests that in some ATL cases, the clonality of monoclonally proliferated HTLV 1 infected cells may be changeable before overt ATL onset. PMID- 8411654 TI - [Pathology of arteriosclerosis--an overview]. AB - Arteriosclerosis is vascular disease characterized by thickening, hardening and remodelling of the arterial wall and classified into following three categories: atherosclerosis, Monckeberg's medial calcific sclerosis, and arteriolosclerosis. Fibromuscular intimal thickening starts its development in the fetal age of the 6th month and continues to grow with aging. The specific topography of early atherosclerotic lesions is primarily attributed to wall shear stress, one of hemodynamic forces. The lesion will proliferate to form atherosclerosis when complicated by hyperlipidemia, hypertension, and/or other clinical risk factors. The major complications of atherosclerosis, stenosis of the arterial lumen and thrombus formation at ulcerated arterial walls, frequently cause such lethal diseases as ischemia of various pivotal organs or rupture of aneurysms. PMID- 8411655 TI - [Molecular biological studies on the pathogenesis of atherosclerosis]. PMID- 8411656 TI - [Recent advances in lipoprotein metabolism]. AB - Recent advances in lipoprotein metabolism were reviewed. The scavenger receptor for acetyl-LDL cloned by Kodama et al. was immunohistochemically demonstrated to exist in the human atherosclerotic lesions. LDL-receptor related protein (LRP) and VLDL receptor which would recognize apoE but not apoB have been recently cloned. Transgenic mice expressing high levels of apolipoproteins such as apoA-I and apoE or apolipoprotein deficient mice produced by gene targetting technique are providing important information on the function of apolipoproteins. PMID- 8411657 TI - [Carbohydrate metabolism]. AB - This review summarizes progress in glycosylation research of relevance to atherosclerosis. Glucose reacts in vivo with cellular proteins nonenzymatically and forms Amadori products. The Amadori products proceed very slowly to undergo a number of further dehydration and rearrangement to form advanced glycosylation end products (AGE). The AGE moiety are characterized by being brown, fluorescent chromophores that can cross-rink proteins. In contrast Amadori products, AGE are irreversible and accumulate on long-lived proteins (eg, collagen, enzyme, lens crystallins) for many years. AGE proteins can modify lipoproteins fibrinogen, collagen and DNA. AGE protein receptor is identified on macrophages. AGE may accelerate development of atherosclerosis by various manner. PMID- 8411658 TI - [The role of collagen matrix in atherogenesis]. AB - The different types of collagen including Types I, III, IV, V, VI and VIII are distributed in the artery. The atherosclerotic lesions, such as fatty streaks and atheromatous plaques, are characterised by an increase in types V and VI, which are localized in the region with a deposition of LDL and an infiltration of macrophages. Vascular smooth muscle cells interact with various types of collagen. Collagen matrices induce an enhancement of differentiation and suppression of cellular proliferation of "normal" smooth muscle cells. However, transformed smooth muscle cells fail to interact with various types of collagen. These results suggest that the interaction between vascular cells, including smooth muscle cells, macrophages and endothelial cells and collagen matrices is important in atherogenesis. PMID- 8411659 TI - [Function of vascular endothelial cells]. AB - Endothelial cells lining on the luminal surface of vessels play important roles in maintaining the integrity of vasculature. Vascular endothelium is known as a metabolically active secretory tissue, presents a thromboresistant surface, and acts as a selective barrier. It is now believed that abnormality in structure and function of endothelial cells is the cause of not only vascular diseases including atherosclerosis but also certain visceral disorders. In this article, I will focus on the regulatory role of vascular endothelial cells in vascular tone, adhesion of leukocytes, thrombosis, fibrinolysis, and cell growth. PMID- 8411660 TI - [Regulation of differentiated properties of arterial smooth muscle cells]. AB - The phenotypic transition of smooth muscle cells (SMC) from a contractile to a synthetic state appears to be an early event in the pathogenesis of atherosclerosis. It was found that plasma fibronectin promotes the transition of rat aortic SMC into the synthetic phenotype and that laminin delays this process. Recently, we found that type I collagen is as efficient as fibronectin in promoting the initial attachment and phenotypic transition of rabbit arterial SMC and elastin maintained the cells in the contractile phenotype, as laminin. On the other hand, laminin plus type IV collagen suggest that a partial remodulation may be possible. These findings emphasize the importance of extracellular matrix components in the control of the differentiated properties of arterial SMC and may be important for clarifying the development of atherosclerosis. PMID- 8411661 TI - [Macrophage scavenger receptors and the maintenance of blood vessels]. AB - Recent molecular cloning of cDNAs for Macrophage Scavenger Receptors (MSR) reveal several unexpected features of these receptors. These MSRs mediate wide range of ligands, especially negatively charged macromolecules. Expression of MSRs are highly specific to macrophages. The intracellular pathways for endocytosis and recycling of MSRs are efficient only in macrophages. Immunocytochemical study of MSRs show that all of the tissue macrophages, including brain scavenger cells (FGP cells) are MSR positive. Under some pathological conditions, such as atherosclerotic lesions, macrophages infiltrating into these lesions, which are derived from monocytes, are also positive for MSR. Macrophages expressing MSR are thought to play an important role in the clearance and maintenance of blood vessels. Brain scavenger cells may play a role in the maintenance of small blood vessels in the brain. PMID- 8411662 TI - [Platelet functions in atherosclerosis]. AB - Recent advances on platelet functions involved in atherosclerosis and subsequent thromboembolic diseases are reviewed. Platelets show multiple roles in hemostasis/thrombosis, wound healing, allergy, inflammation, metastasis of malignant cells and vasospasms through aggregation/secretion reaction. In atheromatous plaque, migration and proliferation of vascular smooth muscle cells and macrophages are the most prominent findings. In this pathological state, platelet may play as an enhancer by the secretion of bioactive substances including platelet-derived growth factor, serotonin and platelet factor 4. Investigation on platelet dynamics must be carried out not only for monitoring but also for the assessment of progressive factors in atherosclerotic disease. PMID- 8411663 TI - [Coagulation and fibrinolytic systems and atherogenesis]. AB - Normal blood fluidity and perpetuation of the non-thrombogenic state are primarily maintained by the anticoagulant and fibrinolytic systems of vascular endothelial cells involving heparin-like molecule, thrombomodulin, prostacyclin, and the receptor for tissue plasminogen activator. Atherosclerosis perturbes these activities, resulting in arterial thrombosis. On the other hand, recent experimental evidence suggests that the disordered thromboregulation often promotes atherosclerosis. Several known risk factors for development of atherosclerosis, including homocysteine and lipoprotein (a) perturb anticoagulant and fibrinolytic systems of vascular endothelial cells at an early stage of atherogenesis. PMID- 8411664 TI - [Antiatherogenic action of omega-3 polyunsaturated fatty acids]. AB - Recently, atherosclerosis has become a matter of great concern because of a rapid increase of aged people. It has been widely known that omega-3 polyunsaturated fatty acids, rich in marine fish oil, have an antiatherogenic action. The mechanism is ascribed to its action on serum lipids, functions of the platelets, endothelial cells, smooth muscle cells and macrophages, prostaglandin and leukotriene metabolism, growth factor and cytokine production, EDRF production, blood pressure, etc. In this paper the basic and clinical evidences concerning the effects of fish oil on these multifactors, which are deeply involved in the development of atherosclerosis are reviewed. PMID- 8411665 TI - [Roles of cytokines and growth factors in atherogenesis]. AB - In lesions of atherosclerosis, various cytokines and growth factors, which are generally not expressed in the normal artery, are upregulated. Several of them including PDGF, bFGF, HB-EGF, IGF-1, IL-1 and TGF-beta and TNF play key roles in atherogenesis by stimulating chemotaxis and proliferation of vascular smooth muscle cells and production of extracellular matrix substances such as proteoglycans, collagen and elastic fibers by those cells. Endothelial cells and macrophages are also the targets as well as the sources of those cytokines and growth factors. The production of those cytokines or growth factors are regulated by molecules of each other or by themselves forming a complex cytokine network. Understanding and control of the roles of those cytokines in vascular walls will provide an insight on the mechanism of atherogenesis and contribute to the development of better ways to its prevention. PMID- 8411666 TI - [Progress in diagnosis of arteriosclerosis in humans--review]. AB - Most of cerebrovascular diseases or ischemic heart diseases, the major causes of death in Japan, occur as terminal symptoms of atherosclerosis. Much effort has been made to prevent these diseases and also to assess premature atherosclerosis. Recently, new models of tools for diagnosis has been developed. There are several ways to determine the existence of atherosclerosis, namely (1) to find the major risk factors of atherosclerosis such as hyperlipoproteinemia, hypertension, diabetes mellitus and/or smoking, and also (2) for apparent cardiovascular patients, to examine the pathogenesis of the disease by using invasive methods such as selective arteriography, intravasculoscopy, and intravascular ultrasonic examination, if necessary, and (3) for asymptomatic people, to apply hypoinvasive examinations such as the ultrasonic examination, the computed tomography and magnetic resonance angiography. As a non-invasive diagnostic method, we have been trying to assess the progression of atherosclerosis clinically by computed tomography (CT) of the abdominal aorta. Recently we developed a new computer program to define the wall thickening and calcification of aorta on personal computer. In the near future, progression and regression collaboration studies in Japan may be achieved. PMID- 8411667 TI - [Angiography in arteriosclerosis]. AB - Advances in angiography of arteriosclerosis have been made possible by improvement of angiographic techniques, materials, such as narrow catheters and gliding guidewires, x-ray apparatus and equipments, including digital subtraction angiography (DSA), computed radiography and magnetic resonance angiography and contrast media. Improved, narrow and yet wide lumen catheters have made it possible to insert the catheter via the axillary or brachial artery. The matrix of DSA was increased to 1024 x 1024 from 512 x 512, and resolution of DSA has been greatly improved. Intravenous DSA has made examination of aneurysm or arterial stenosis or occlusion, easier without much harmful invasion. Ionic or non-ionic low osmolar contrast media has greatly reduced the irritation to the vessels. PMID- 8411668 TI - [Evaluation of arterial sclerosis by CT, MR imaging and MR angiography]. AB - Wall thickening and intimal changes obtained by enhanced CT are early findings of aortic sclerosis. These findings are often detected in the lower abdominal aorta and middle thoracic descending aorta of normal subjects over 30 years of age, as predictors of atherosclerosis. Arterial calcification is a useful sign for evaluating atherosclerosis, especially, in the coronary arteries. The sensitivity and specificity of CT-detected coronary calcification in coronary stenosis by CAG were 76% and 80%, respectively, in our study. In the patients with atherosclerosis, MRI demonstrates wall thickening, intimal projection and narrowing of the lumen in the aorta and large arteries. MRA can be applied to more peripheral arteries, particularly, in the head and extremities, and will be available for screening of atherosclerotic disease in the near future. PMID- 8411669 TI - [Transesophageal echocardiography in atherosclerosis]. AB - CT scan and MRI were used for evaluating the degree or extent of atherosclerotic lesion. However, these methods do not show characteristics of the aortic wall and the hemodynamics surrounding atherosclerotic lesion. Recently, evaluation of the severity of aortic atherosclerosis, especially the thoracic aorta, and the proximal portion of the coronary artery, became possible by transesophageal echocardiography. This article reviews the diagnosis of aortic atherosclerosis and coronary atherosclerosis, using transesophageal echocardiography. PMID- 8411670 TI - [Doppler ultrasound in the diagnosis of atherosclerosis]. AB - Atherosclerosis causes various types of flow disturbances within the affected arteries. Recent advances in the Doppler imaging technique have made possible evaluation of the temporal and spatial flow characteristics in the different portions of the arterial system, such as aorta, coronary, carotid and peripheral arteries. In the diagnosis of dissecting aneurysm of the aorta, transesophageal Doppler color flow imaging is quite useful to detect the site of entry, which is essential information for surgery. Intracoronary flow velocity can be accurately measured by intravascular Doppler technique. The recent introduction of a Doppler tipped angioplasty guide wire makes it possible to measure the phasic flow deep within a coronary artery. The flow velocity measurement using a duplex system enables us to quantify the stenotic severity in carotid and peripheral arteries, and the Doppler color flow imaging, which has become available recently, also facilitates the demonstration and grading of atheromatous lesions in these arteries. PMID- 8411671 TI - [Intravascular ultrasound (IVUS) in the diagnosis of atherosclerosis]. AB - Intravascular ultrasound (IVUS) is a newly developed diagnostic modality for atherosclerosis. Early in vitro studies showed that IVUS could identify normal vessel structures and atherosclerotic lesions, such as intimal thickening, lipid rich plaque, fibrous plaque, and calcification. In clinical settings, IVUS offers precise quantitative measurement of vascular stenosis as well as providing new information regarding the transmural distribution and the qualitative assessment of vascular atherosclerotic lesions. IVUS is also useful for assessing the results and the mechanisms of catheter-based interventional therapeutic devices, such as balloon angioplasty, laser angioplasty, stent, and atherectomy. The development of three-dimensional reconstruction of ultrasound images would enhance the diagnostic quality and afford a reliable guidance for interventional therapeutic procedures. PMID- 8411672 TI - [Radionuclide studies of atherosclerosis]. AB - New Tc-99m-labeled brain perfusion imaging agents, such as Tc-99m-HMPAO, and Tc 99m-ECD, are developed and expected to improve the clinical study of cerebral perfusion disturbance due to arteriosclerosis. They enable us better images than I-123-IMP and more accurate diagnosis. Similarly, in the field of cardiovascular nuclear medicine, new Tc-99m-labeled compounds, Tc-99m-MIBI, Teboroxime and Tetrofosmin are available. Furthermore, I-123-MIBG, a tracer taken up in presynaptic adrenergic vesicles, and I-123-BMIPP, a new agent developed for the evaluation myocardial fatty acid metabolism, will mark a new epoch in the study of coronary sclerosis. In order to image atherosclerosis directly, In-111 polyclonal immunoglobulin G, Tc-99m-low density lipoprotein etc., are under investigation. PMID- 8411673 TI - [Angioscopic evaluation of atherosclerosis]. AB - Recently, percutaneous transluminal angioscopy has been clinically used. We describe the evaluation of atherosclerosis by angioscopy. In observation of aortic and peripheral arteries, color of the luminal surface is yellow in the lesion without endothelial fibrosis, whereas, it is white with endothelial fibrosis. A variety of findings such as endothelial flap, endothelial thickening, mural bleeding, mural or occlusive thrombi are observed in the atherosclerotic lesions. It is difficult to detect these findings by angiography. In observation of coronary arteries, color of the luminal surface is yellow or white and often associates spiral folds in the early stage of atherosclerosis. Plaques are classified to two categories. One is the complex plaque characterised by endothelial flap, rupture of atheroma, ulceration, mural thrombus and usually observed in patients with unstable angina or myocardial infarction. Another is the regular plaque without those findings and observed in patients with stable angina. Angioscopy is useful for evaluation of atherosclerosis. PMID- 8411674 TI - [Pulse wave velocity, thermography in the diagnosis of arteriosclerosis]. AB - 1. Pulse wave velocity (PWV) increased with increasing organic stiffness of the arterial wall mainly caused by arterio-sclerosis and also increasing functional stiffness of the arterial wall mainly caused by elevation of blood pressure. Age related changes in PWV have been recognized in variable countries. The PWV in the ascending aorta calculated by the characteristic impedance (378 +/- 141 cm/sec) was lower than that of conventional measurement in the descending aorta (697 +/- 144 cm/sec). 2. Thermography provide us indirect information of arteriosclerosis through reduction of skin temperature caused by disturbance of blood flow. PMID- 8411675 TI - [Progress in the treatment of atherosclerotic vascular diseases]. AB - The treatment of atherosclerotic vascular diseases has improved greatly. Clinical and epidemiological studies have recently revealed the following data. Changes in lifestyles is extremely effective and should be prevailed widely. Amelioration of atherosclerosis by drugs, including hypolipidemics, antihypertensives and antioxidant agents, is also confirmed. Skills and devices are much improved in PTCA and DCA which face the possibility of restenosis. Control of risk factors is also considered to be important to prevent restenosis. PMID- 8411676 TI - [Dietary treatment of atherosclerosis]. AB - Dietary treatments for hyperlipidemia, hypertension, diabetes mellitus and obesity are essential for the prevention or management of atherosclerosis. To correct overweight or obesity, restriction of energy intake should be considered. The consumption of fat should be decreased less than 25 percent of total energy. The ratio of saturated fatty acid, monounsaturaled fatty acid and polyunsaturated fatty acid is recommended as 1:1:1. Intake of oily fish should be included in daily meal. The ratio of n-6/n-3 polyunsaturated fatty acid is considered to be beneficial around three or four. Excess intake of simple sugars must be avoided and increase the intake of complex carbohydrate and dietary fibre are recommended. PMID- 8411677 TI - [Exercise therapy of atherosclerosis]. AB - The effect of exercise on the progression of atherosclerotic diseases is reviewed. Prospective studies have reported that a low daily physical activity is associated with a lower incidence of coronary artery disease. Mild aerobic exercise improves survival in patients with coronary artery disease, lowers blood pressure in patients with mild hypertension, improves insulin resistance in diabetes mellitus, lowers weight in obesity and increases HDL-cholesterol and decrease triglyceride in patients with dyslipidemia. Mild exercise therapy may be beneficial in the management of atherosclerotic diseases. PMID- 8411678 TI - [The treatment of atherosclerosis--drug therapy]. AB - Drug treatment against atherosclerosis has been evaluated recently in many epidemiological studies. Lipid Research Clinics Group convincingly reported in a large scale design that anion exchange resin effectively reduced blood cholesterol level and concomitantly decreased the events of coronary heart disease. Subsequently, anion exchange resin with or without combined administration of niacin or statin was found to inhibit the progression of coronary atherosclerotic lesions in FATS, SCOR, CLAS and STARS. Fenofibrate also successfully reduced the coronary artery narrowings. Based on these intervention studies, several hypocholesterolemic agents are definitely effective in the treatment of coronary atherosclerosis. PMID- 8411679 TI - [Percutaneous transluminal angioplasty in atherosclerosis]. AB - Percutaneous transluminal balloon angioplasty has achieved a dominant role in the interventional treatments of atherosclerotic peripheral, renal, and coronary artery disease. Improved operator's technique and equipment design have increased the primary success rate. Despite improved success rate and safety, acute closure, late restenosis and difficulty in treating chronic total occlusions or diffuse lesions remain as serious limitations of this procedure. To overcome these limitations of balloon angioplasty, new devices such as stenting, atherectomy or laser ablation have been developed. Although there are many problems for each device, these techniques appear to reduce the limitations in angioplasty. PMID- 8411680 TI - [Surgical treatment of arteriosclerotic arterial occlusive disease]. AB - Various kinds of surgical technique have been utilized for vascular reconstruction for arteriosclerotic arterial occlusive disease (ASO). In general concept, application of thromboendarterectomy is limited to the segmental occlusive aortoiliac atherosclerosis and bypass surgery can be used for almost all types of arterial occlusion. Patency result after bypass surgery are well established and the five year patency rates reveal more than 90% in aortoiliac disease, 75-80% in femoropopliteal disease, and around 70% in crural reconstruction respectively. Despite of these recent advances of reconstructive vascular surgery, there have been augmented several controversies in surgical treatment of ASO. How to treat multisegmental lesion which is characterized by high incidence of ischemic ulcer or gangrene. What is the treatment of choice for high risk patient especially with ischemic heart disease. In Japan, aged patient over 80 have been increasing and they are frequently complicated with limb threatening ischemia and usually grouped into high risk patient. Endovascular intervention, which is developed under the basis of recent technology, is the one of the topics of treatment of peripheral vascular disease. Its long term results, however, are not sufficiently evaluated and how to select the proper reconstructive procedures including the endovascular intervention for high risk patient is remained to be controversy. PMID- 8411681 TI - [Cerebrovascular diseases due to extra- and intracranial atherosclerosis]. AB - (1) In Japan, atherosclerosis in the extracranial arteries has now been more or less prevailing. (2) Many devices are widely used to extra- and intracranial vascular disease. However, such a simple sign as an auscultation of the carotid bruit or aortic-arch calcification in the X-ray picture may give valuable information of extra- and intracranial atherosclerosis. (3) Anti-heat-shock protein 64 antibody has recently been presented as an independent risk factor of carotid atherosclerosis. (4) Clinical features of carotid occlusion or stenosis are markedly variable. PET study has revealed that a water-shed ischemic zone may be in a state of "misery perfusion". (5) Regression may occur in the cervical as well as cerebral atherosclerosis. PMID- 8411682 TI - [Ischemic heart disease]. AB - During the progression of early atherosclerotic lesions, atherosclerotic plaque rupture, with intraluminal thrombosis superimposed, which is one of the principle mechanisms of evolving atherosclerosis, may lead to thrombotic occlusion and ischemic coronary syndrome. Pathologic studies suggest that plaque rupture and overlying thrombi, which are dynamic and repetitive may frequently occur. In most cases, healed ruptures and incorporation of thrombi produce plaque progression without manifestation of clinical symptoms. However, acute mural occlusive thrombi, overlying plaque ruptures, cause unstable angina and acute myocardial infarction. The different pathogenesis between unstable angina and acute myocardial infarction might depend on the composition and stability of the thrombus resulting from the degree of vessel injury and blood flow. Soft lipid rich plaques appear more prone to rupture, particularly when the lipid pool is localized eccentrically within the intima. Macrophages in the plaque may also facilitate plaque rupture by releasing proteases. PMID- 8411683 TI - [Nephrosclerosis: current status]. AB - Nephrosclerosis is the most typical and widespread renal manifestation of hypertension and can be judged as the pathological hallmark of essential hypertension. Nephrosclerosis is an important and frequent cause of progressive renal disease, however, information in the literature on the risk of developing renal failure in the course of essential hypertension is sparse. Traditionally, nephrosclerosis was thought to result from glomerular ischemia. Alternatively, glomerular sclerosis in hypertension may result from glomerular hyperperfusion or hypertension. Studies in experimental models of renal disease have identified a promising intervention with either Ca antagonists or angiotensin-converting enzyme inhibitors. Application of these therapies to patients with nephrosclerosis should await the results of careful clinical trials. PMID- 8411684 TI - [Ischemic bowel disease]. AB - Arteries of the small intestine and colon have several anastomotic interconnections that help protect against the consequence of occlusive vascular disease. Vascular ischemic syndromes of the colon include several clinical entities, depending upon the degree and duration of the interruption of vascular flow. They include acute mesenteric artery occlusion, abdominal angina and ischemic colitis. Acute mesenteric artery occlusion is usually caused by an embolus from the heart and if the diagnosis is delayed, the mortality is quite high. Abdominal angina often precedes the acute mesenteric artery occlusion. On the other hand, ischemic colitis consists of the solitary disease entity because of its characteristic clinical and radiographic features. Recently, many cases of ischemic colitis have been reported. This article reviews the recent advances in ischemic bowel disease. PMID- 8411685 TI - [Aortic aneurysm]. AB - Atherosclerotic true aortic aneurysms develop in the abdominal aorta more commonly than in any other arteries. Once developed, the natural course of true aortic aneurysm is one of progressive growth until it ruptures. The growth rate of true aortic aneurysms is variable, but the average is approximately 1.5 mm in diameter in the thoracic, and 4 mm in the abdominal aorta per year. The symptoms of aneurysms are variable. Many true aneurysms are asymptomatic, but usually the onset of chest or back pain develop in aortic dissection. Medical imagings, such as ultrasound and CT scan, are the most useful diagnostic methods for aortic aneurysms. And preoperatively, atherosclerotic lesions in the other arteries, such as coronary artery or cerebral artery, that cause perioperative death or late complications must be studied. PMID- 8411686 TI - [Arteriosclerosis obliterans]. AB - Arteriosclerosis obliterans (ASO) has become one of major health problems in the elderly people in Japan. This paper reviews the recent progress in the diagnosis and treatment of ASO. Both medical and surgical treatments, such as atherectomy catheters, endovascular metallic stenting, laser angioplasty and LDL-apheresis have recently been greatly developed. As atherosclerosis is a generalised disease, patients with ASO tend to have other atherosclerotic diseases, i.e., cerebrovascular disease, coronary heart disease and renal vascular disease. For the selection of treatments, we have to consider the whole background of the patients, including a quality of life. PMID- 8411687 TI - [Role of hypertension in atherosclerosis]. AB - Hypertension is frequently associated with clinical cardiovascular complications due to atherosclerosis. However, several previous studies have showed that incidence of cardiovascular complication due to atherosclerosis hypertensives do not decrease even when the blood pressure was well controlled. The effects of hypertension on atherogenesis appears to be mainly due to two factors: (1) The mechanical stress which induces the change of cell functions of arterial wall cells such as endothelial cells, smooth muscle cells and extracellular matrix, (2) Vasoactive substances, such as angiotensin and endothelin, which may plays a role for smooth muscle cell proliferation. The tropic changes in smooth muscle cells induced by above two factors in hypertension might reflect common alternations in cellular reactivity as observed in atherosclerosis. If so, hypertension and atherosclerosis could be regarded as two effects with one underlying cause. PMID- 8411688 TI - [Diabetes mellitus as a risk factor for arteriosclerosis]. AB - Diabetes mellitus is an important risk factor for coronary heart disease and cerebral vascular disease. Moreover, diabetic patients are frequently hyperlipidemic (our recent study has revealed that approximately 70% of diabetic patients are dyslipidemic) and are at a higher risk when complicated with hyperlipidemia. In addition, abnormalities in lipoprotein metabolism, which may promote arteriosclerosis, are seen even in normolipidemic diabetics. One such alteration is an increased cholesterol level in intermediated-density lipoproteins from normolipidemic diabetic patients. Recently, abnormal particles in low-density lipoprotein (LDL) fraction, glycated-LDL and small, dense LDL, are regarded as atherogenic lipoproteins. Several factors promoting arteriosclerosis in diabetics, such as abnormal lipoprotein metabolism, increase of lipoprotein (a), hyperinsulinemia, enhancement of polyol pathway, dysfunction of platelet, are described here. PMID- 8411689 TI - [Hyperuricemia and atherosclerosis]. AB - To clearly determine whether hyperuricemia participates directly in atherosclerotic disease or not, the prognosis and associated factors were studied, based on data from 104 patients whose serum uric acid had been completely maintained at normal levels with prolonged medication. The mean age at death was 65.8 +/- 10.5 years. The causes of death were as follows: cardiovascular disease (26.9%), cerebral disease (26.2%), malignant neoplasms (26.0%), uremia (7.6%), and miscellaneous disease (18.3%). Serum lipids especially triglycerides, body weight and influenced on the prognosis of the patients FBS. Most common complications were in the cardiovascular disease group; hypertension and hyperlipidemia. These data suggested that the apparent increased incidence of cardiovascular disease in gout rather than renal failure bore a relationship to such complications as hypertension or hypertriglycemia. Hyperuricemia alone may not be an atherosclerotic risk factors. There was no correlation between treatment with allopurinol and probenecid and cardiovascular complications. PMID- 8411690 TI - [Primary hyperlipoproteinemia]. AB - Hyperlipidemia is first detected by an increase in the plasma concentrations of cholesterol and/or triglycerides, and implies an abnormality of plasma lipoprotein metabolism. Disorders of lipoprotein metabolism are often classified specifically according to the lipoprotein affected. The WHO classification of lipoprotein phenotypes is a useful means of showing which lipoproteins are present in excess in individual hyperlipidemic patients. Hyperlipoproteinemia can be secondary to other well-known diseases that affect plasma lipoprotein metabolism, for example, diabetes mellitus, hypothyroidism or nephrotic syndrome. When such diseases are excluded, the hyperlipoproteinemia is defined as primary hyperlipoproteinemia. Many primary hyperlipoproteinemias have a genetic basis and the underlying molecular defect has been clarified in some genetic disorders. Hyperlipoproteinemia is considered to be one of the major risk factors for atherosclerosis and the development of atherosclerosis depends on the type of hyperlipoproteinemia. In this sense, familial hypercholesterolemia is a clinically important primary hyperlipoproteinemia because of its high risk of ischemic heart disease and its high prevalence in a normal population (1/500). It is necessary to make an exact diagnosis of specific genetic disorder, if possible, to provide prognostic and therapeutic information. PMID- 8411691 TI - [Atherosclerosis and endocrine disorders]. AB - It has been demonstrated that high cholesterol levels are correlated with development of atherosclerosis, while high levels of high density lipoprotein (HDL) are associated with reduced cardiovascular mortality. Some endocrine disorders accelerate atherosclerosis in association with hypercholesterolemia, hypertension, low level of HDL and hypertriglyceridemia. In patients with acromegaly, hypertriglyceridemia is sometimes accompanied with and aggravated by the presence of impaired glucose tolerance. In patients with hypothyroidism, coronary atherosclerosis may develop in association with hypertension, hypercholesterolemia and moderate elevation of triglyceride which is often accumulation of intermediate lipoprotein. Cushing syndrome may accelerate atherosclerosis by the fact that corticosteroid may induce or exacerbate several known coronary risk factors including hypertension, hypercholesterolemia and impaired glucose tolerance. Estrogen has beneficial effects on the cardiovascular system for postmenopausal women by affecting lipid metabolism, decrease of LDL and increase of HDL. PMID- 8411692 TI - [Atherosclerotic and hemodynamic effects of obesity]. AB - Obesity is a risk factor of atherosclerosis. The TG content of a fat cell is determined by the balance of lipogenesis from plasma FFA and glucose and lipolysis by hormone-sensitive lipase (HSL). Plasma FFA is produced by TG lipolysis by lipoprotein lipase (LPL). Insulin stimulates LPL activity and inhibits HSL activity. Therefore, hyperinsulinemia stimulates TG accumulation in fat cells. Insulin also stimulates fat cell proliferation. Hyperinsulinemia is a major factor for obesity. Portal FFA stimulates VLDL synthesis and gluconeogenesis and inhibits insulin degradation in the liver. Therefore, visceral obesity is important as a risk factor of atherosclerosis. However the increase of total adipose tissue mass is more important for blood pressure and cardiac performance. PMID- 8411693 TI - [Molecular genetical approach to neurological diseases]. PMID- 8411694 TI - [Positional cloning--current status and future directions]. AB - Several strategies for isolating disease genes have been developed. In functional cloning, the gene is isolated based on information on the function of gene products (proteins). In contrast, positional cloning is based on the isolation of the disease gene, starting from the knowledge of its location on the genome. The flow chart of positional cloning was described as follows; 1) identification of the location of the disease gene by linkage analysis. 2) construction of the physical map spanning the region. 3) isolation of the candidate genes from the region. 4) identification of mutations in the candidate genes, which is specific to the disease. The current concept and strategies on the positional cloning is discussed. PMID- 8411695 TI - [Application of the chromosome sorting technique to the human genome analysis]. AB - The flow-sorted human chromosome is an ideal DNA source for genome analysis research, particularly gene mapping and construction of cosmid library. For gene mapping, 50,000 chromosomes each were sorted on a membrane filter and hybridized with a radio-labeled gene probe. Using this Spot Blot Hybridization method in combination with FISH, we have mapped more than 30 new genes including 6 neuron specific ones. For construction of a chromosome-specific cosmid library, nearly 10 million chromosomes were flow-sorted. The chromosome 21-specific cosmid library consisting of 33,000 clones and the 22pter-q11 region-specific library consisting of 9,200 clones were established. Grouping of cosmid clones from the library using high density replica filters and YAC DNAs to make cosmid contigs within specific regions is under way. PMID- 8411696 TI - [Laser chromosome microdissection and cloning of the genetic disease loci]. AB - We have developed an argon ion laser chromosome microdissection technique in conjunction with a single unique primer polymerase chain reaction (SUP-PCR) to directly amplify microdissected chromosomes. The 22-mer primer used in PCR, although unique in sequence, randomly primed and amplified any target DNA. These methods were applied to both the terminal region of the human chromosome 4p (4p 16) and Xq (Xq26-q28), and two chromosome region-specific DNA libraries were constructed. The resulting libraries contained approximately 1000 nonoverlapping DNA sequences with an average size of 230-350 bp, at an average spacing of 10-65 Kbp along the chromosomes of origin. Our new method is a simple and general approach for constructing a chromosome region-specific DNA library from a single metaphase spread. PMID- 8411697 TI - [Structural analysis of human genome by YAC technologies]. AB - A method for construction of YAC (Yeast Artificial Chromosome) libraries with large inserts has been developed and promoted the ongoing project of human genome. Isolation by PCR screening charges the YAC clone with a unique tag of a pair of PCR primers at the defined chromosome site (Sequence Tagged Sites; STS). Current evaluation of YAC has revealed that larger YAC has more problems where rearrangements including deletion and chimera occur extensively in DNA molecules, presenting a limited use of this technology in mapping; the contig map with mega YACs will be substituted by some other system such as cosmids with which the human healthy and disease genes will be characterized. PMID- 8411698 TI - [Physical mapping of human chromosomes with linking clones and jumping clones]. AB - Physical maps facilitate the correlation of genetic linkage maps and actual genome structure, and contribute to the immediate access to disease genes in question. The introduction of pulsed-field gel electrophoresis (PFGE) made it possible to construct restriction maps in Mb order with rare-cutting enzymes. Linking clones and jumping clones are useful tools for constructing long-range and accurate maps. The linking clones are used to identify adjacent restriction fragments and the jumping clones are to identify neighboring restriction sites. The general strategy for physical mapping using these clones and our application to NotI restriction mapping of human chromosome 21 are described. PMID- 8411699 TI - [New developments of molecular genetic analysis on human mitochondrial DNA]. AB - The use of magnet beads for the capture and purification of PCR product allows simple preparation of immobilized single-stranded template. This template is ideal for use in DNA direct sequencing. The end labeling of PCR product with biotin is achieved simply by using a biotinylated primer. Capture and purification are then accomplished by incubation with Streptavidin attached magnet beads and magnet separation. Once immobilized, the DNA is converted to a single-stranded template by elution with alkali and another magnetic separation. Immobilized single-stranded template prepared in this manner are suitable for manual and automated solid-phase DNA sequencing by the dideoxynucleotide chain termination method. A 100% single-stranded template is produced, without interference from primers, free nucleotides or the complementary strand. The solid phase approach provides reproducible sequencing results with high yields. The eluted complementary strand can also be sequenced after neutralization. PMID- 8411700 TI - [Lysosomal storage disease: a group of genetic neurodegenerative disorders]. AB - Lysosomal storage disease is a group of neurometabolic diseases mainly occurring in infancy and childhood. They were first recognized as new diseases on the basis of unique clinical manifestations or pathological findings, and then the stage of biochemical analysis of storage material and enzyme assays in tissues and cells from patients followed. Recent technological development has enabled us to look further into the molecular genetic basis of these inherited diseases. Protein analysis revealed intracellular events of the mutant enzyme molecule responsible for the pathogenesis of a disease, and more detailed information has been obtained about the mutant gene and its product. Clinical manifestations are not always uniform for a single disease with mutations in the same gene. Clinical subtypes have been proposed for many lysosomal diseases. At present, the molecular and metabolic basis of each phenotypic expression is not clear, although common mutations have been found for specific clinical forms in some diseases. In this article, the current status of lysosomal disease research was summarized, particularly focusing on molecular pathology and molecular diagnosis. Finally future prospects for pathogenetic analysis of neural dysfunction and possible gene therapy were briefly discussed. PMID- 8411701 TI - [Molecular genetics of beta-galactosidase deficiency (GM1-gangliosidosis and Morquio syndrome type B)]. AB - Recent advances in the molecular study of beta-galactosidase deficiency (GM1 gangliosidosis and Morquio syndrome type B) are reviewed. Until now, 14 different mutations have been found in the beta-galactosidase gene in patients with this disorder. Gene mutations are heterogeneous, but common and specific mutations have been identified for three types of protracted clinical course; 51Ile-->Thr mutation for Japanese adult/chronic GM1-gangliosidosis, 201Arg-->Cys for Japanese late infantile/juvenile GM1-gangliosidosis and 273Trp-->Leu for Caucasian Morquio syndrome type B. These phenotype-specific mutant genes produce mutant proteins with significant residual enzyme activity, whereas mutant proteins associated with infantile GM1-gangliosidosis patients show complete loss of enzyme activity. The phenotypic variations of this disorder may be related to different mode of intracellular processing and turnover of mutant enzyme proteins. PMID- 8411702 TI - [Lysosome disease--Sandhoff disease]. AB - Lysosomal beta-hexosaminidase occurs as two major isozymes hexosaminidase A and B. The alpha subunit is encoded by the HEXA gene and the subunit by HEXB gene. Defects in the beta subunit lead to Sandhoff disease. Patients with the defect lack the activity or formation of both hexosaminidase A and B. The disorders are classified according to the age of onset, as infantile, juvenile and adult form. Recent molecular genetic analysis has revealed a 50 kb deletion, 16 kb Alu type deletion, and compound heterozygous with other mutations. In the juvenile or adult type of the disease, point mutation of the HEXB gene, creating a new 3' splice acceptor site. The correlation of the clinical phenotype and the gene abnormalities is discussed. PMID- 8411703 TI - [Tay-Sachs disease]. AB - GM2-gangliosidosis is a group of neurological disorders resulting from genetically defective catabolism, and consequent abnormal accumulation, of GM2 ganglioside. Three major types are distinguished: the B variant (Tay-Sachs disease), the O variant (Sandhoff disease), and the AB variant, caused by genetic abnormalities in the genes coding for the beta-hexosaminidase alpha- or beta subunit, or the GM2-activator protein, respectively. A number of gene abnormalities responsible for Tay-Sachs disease have already been identified and the correlation between the beta-hexosaminidase alpha gene abnormality and the clinical phenotype has been explained in many cases. In the severest phenotype of Tay-Sachs disease (infantile form), mRNA of beta-hexosaminidase alpha subunit is not produced or is unstable such as in French Canadian patients or in Jewish patients with infantile Tay-Sachs disease, or the polypeptide does not have any catalytic activities because of the alteration of glycosylation such as the mutation of Glu482-to-Lys found in a Italian patient or the altered structure of polypeptide. The mutation identified in a large majority of the Japanese infantile Tay-Sachs disease patients, which is a G-to-T substitution at 3'-end of intron 5, generates a short mRNA with complete skipping of exon 6 and a polypeptide lacking 34 amino acids is generated but catalytically inactive. On the other hand, some active alpha beta dimers must be generated in patients with milder phenotypes of Tay-Sachs disease such as Gly269-to-Ser mutation in an adult form. Some of the mutations appear in high frequency among certain ethnic groups such as Ashkenazi Jewish patients and French Canadians.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411704 TI - [Molecular genetic analysis of a Japanese family with Fabry disease]. AB - A Japanese male patient with Fabry disease who had no activity of the lysosomal hydrolase alpha-galactosidase A (alpha-GalA) and female members of his family were analyzed. We cloned a cDNA encoded the mutant alpha-GalA, determined its nucleotide sequence, and found two nucleotide differences between the mutant and the wild-types cDNAs. The one difference, a C-to-T transition at nucleotide number 118, resulted in an amino acid substitution of Pro-40 by Ser. A transient expression assay demonstrated that this missense mutation was the cause of the deficiency of alpha-GalA activity in the patient. Gene analysis of the patient's family by PCR and subsequent sequencing demonstrated that all females were heterozygotes. PMID- 8411705 TI - [Advances in molecular genetics of the Niemann-Pick group of diseases]. AB - Recent advances in molecular genetics of the Niemann-Pick group of diseases are reviewed. Types A and B Niemann-Pick disease are characterized by a deficiency of one of lysosomal hydrolases, i.e. acid sphingomyelinase. The enzyme was partially purified from a large amount of urine and the cDNA clones, encoding acid sphingomyelinase, were cloned. The gene encoding the enzyme has been localized at the region p 15.1-p 15.4 of chromosome 11 by analysis of a somatic cell hybrid and in situ hybridization. Several mutations, causing deficient sphingomyelinase activity, were identified among patients with different ethnic backgrounds. The expression experiments revealed that the mutations responsible for type A cause no detectable residual enzyme activities, while mutations responsible for type B, cause relatively higher residual enzyme activity of 2% to 40 %. Biochemical abnormalities in Type C Niemann-Pick fibroblasts are characterized by normal acid sphingomyelinase activity, accumulation of intracellular cholesterol and defective esterification of exogenously added cholesterol. The basic defect is still unknown. Similar abnormalities were observed in mutant mouse strains, BALB/c and C57 BL/Ks. The mutant C57BL/Ks mouse, which was found in Japan, has been characterized as a sphingomyelinosis and the genetic locus, spm, has been assigned to chromosome 18. By transferring a single human chromosome to the immortalized cell line, we have found human chromosome 18 can reduce intracellular cholesterol accumulation. More recently, Pentchev and co-workers found linkage of type C to human chromosome 18. It is highly probable that the spm and human type C mutations involve the same gene. Molecular cloning of the defective gene in muman and mouse mutation become practically possible. PMID- 8411706 TI - [Molecular diagnosis and gene therapy for Gaucher disease]. AB - Gaucher disease is the most prevalent lysosomal storage disease. It is caused by deficient activity of a lysosomal enzyme known as glucocerebrosidase, also called glucosylceramidase, resulting from mutations in the gene encoding the enzyme. The numerous mutations in glucocerebrosidase gene from patients were reported and the correlation of phenotype and genotype were studied. However, a given genotype cannot be uniquely correlated with a specific phenotype. In the therapeutic point of view, two successful treatment were developed based on the correction of the enzyme deficiency in macrophages. These are bone marrow transplantation and enzyme infusion therapy. Although patients have responded to these two therapies, inherent problems limit their application. Thus, these problems makes Gaucher disease an excellent candidate for therapy based on gene transfer to hematopoietic cells. We succeeded in efficient transduction and sustained high expression of glucocerebrosidase gene in mouse hematopoietic stem cells and macrophages from long term reconstituted mice. The results of our study strengthen the rationale for gene therapy as a treatment for Gaucher disease. PMID- 8411707 TI - [Molecular analysis of the aspartylglucosaminidase gene in Japanese patients with aspartylglucosaminuria]. AB - Molecular analysis of the aspartylglucosaminidase (AGA) gene was performed on two Japanese siblings with aspartylglucosaminuria (AGU). They were the first Japanese patients diagnosed as AGU and up to now, no other patients with AGU have been discovered in Japan. AGA cDNA from one patient contained 7-nt (TCTCCAG) insertion between exons 3 and 4, which was identical with the 3'-terminal sequence of intron 3. Sequencing of amplified genomic DNA from both patients showed the same single base transition (A-->G) at the 5'-side adjacent to the insertion sequence. This mutation newly created a consensus AG dinucleotide at the 3'-splice site and caused alternative splicing of AGA pre-mRNA by leaving the 3'-terminal 7-nt of intron 3. BsmAI restriction site analysis of amplified genomic DNA indicated that they were homozygotes of this mutation. We conclude that splicing defect of intron 3 caused AGA deficiency in them. PMID- 8411708 TI - [Genetic advances in galactosialidosis]. AB - Galactosialidosis is an autosomal recessive inherited metabolic disorder induced by the deficiency of beta-galactosidase and neuraminidase. It can be classified into the early infantile form, the late infantile form, and the juvenile/adult form, by clinical characteristics. This disease has been known to be caused by the lack of protective protein. The human protective protein is synthesized as a 54 kD precursor and then processed to the mature form, a heterodimer of 32 and 20 kD polypeptides. The mature protective protein forms a complex with beta galactosidase and neuraminidase, stabilizing beta-galactosidase and activating neuraminidase. Recently, this protective protein was found to have other multiple functions including activities of carboxypeptidase, esterase and deamidase. The nature of abnormality of the protective protein in the three subtypes of galactosialidosis however has not yet been well elucidated. On the other hand, a cDNA of the protective protein was cloned, and point mutations in the protective protein gene were found in a Japanese family with the adult form, and in Canadian and Italian patients with the late infantile form. We also detected the same point mutation in two Japanese patients with the adult form. Discovery of the genetic defect in different subtypes of galactosialidosis will contribute to the study on the nature of abnormality in the protective protein itself. PMID- 8411709 TI - [Pompe's disease--acid alpha-glucosidase deficiency--a review]. AB - The importance of the role of acid alpha-glucosidase in the lysosomal degradation of glycogen has been emphasized because the deficiency of this enzyme in glycogenosis type II causes glycogen accumulation in lysosomes. Three clinical variants are distinguished. The infantile type has its onset shortly after birth and is known as generalized glycogen storage disease. The adult variant manifests itself mostly after the second decade of life and is characterized by progressive skeletal muscle weakness. The other is childhood type which is usually fatal by the second decade of life. Many biochemical reports of acid alpha-glucosidase have been published. Martiniuk et al reported the cDNA and amino acid sequence of human acid alpha-glucosidase. In prior studies, they reported that the lysosomal acid alpha-glucosidase was polymorphic with three alleles. The rarer allele GAA2 allozyme had a lower affinity for glycogen and starch. We also reported the enzyme heterogeneity in its affinity to Sephacryl S-200 gel. Whereby the enzyme separated into two fractions, S1 and S2. Each fraction contained 76 kDa and 67 kDa components on SDS/PAGE. The spleen enzyme consisted mainly of S1 fraction, containing only a 76 kDa component. In previous extensive studies, different mutations of Pompe's disease have been inferred from alterations in biochemical parameters. More recently Martiniuk et al, Hoefsloot et al and Van der Ploeg et al reported the analysis of cDNA and mRNA. These studies have revealed an absence or abnormal size of mRNA in large numbers of patients and altered restriction endonuclease fragments in a few patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411710 TI - [Mucopolysaccharidosis type VII: characterization of exonic point mutations and molecular heterogeneity]. AB - We identified two different exonic point mutations causing beta-glucuronidase (beta G1) deficiency in three Japanese patients with mucopolysaccharidosis type VII. The beta G1-specific mRNA levels were normal. Sequence analysis of the full length mutated cDNAs showed C-->T transitions, which resulted in a single Ala619- >Val change (case 1, a 8-year old female and case 2, a 24-year old male) and a Arg382-->Cys change (case 3, a 7-year-old female). Each of these two amino acid changes reduced the beta G1 activity of the corresponding mutant beta G1 expressed following transfection of COS cells with expression vectors harboring the mutated cDNAs. PMID- 8411711 TI - [Molecular biology of peroxisome biogenesis]. AB - Molecular mechanisms of peroxisomal protein translocation and peroxisome assembly have been extensively studied these several years. One type of topogenic signals has been identified both in vivo and in vitro: the C-terminal -Ser-Lys-Leu-COOH (SKL) motif sequence. In patients with generalized peroxisomal disease such as autosomal recessive, cerebrohepatorenal Zellweger syndrome where peroxisomes are morphologically absent, all peroxisomal proteins appear to be normally synthesized but assembly of peroxisomes is impaired. We have isolated several somatic animal cell mutants defective in biogenesis of peroxisomes. By genetic complementation analysis following the transfection of cDNA library to a CHO cell mutant, Z65, we cloned a cNDA for 35-kDa peroxisome assembly factor-1 (PAF-1) essential for peroxisome assembly. Furthermore, we have recently delineated the primary defect in a Zellweger patient who belonged to the same complementation group as the Z65. The cause of this syndrome was a homozygous nonsense point mutation in the PAF-1 gene. PMID- 8411712 TI - [PMP70, the 70-kDa peroxisomal membrane protein: a member of the ATP-binding cassette transporters]. AB - We have isolated the cDNA of the 70-kDa peroxisomal membrane protein (PMP70) from rat and human liver cDNA libraries. The nucleotide sequence of the cDNA of PMP70 contains an open reading frame of 1977 bp which encodes an amino acid sequence of 659 residues. Possible two domains were identified by hydropathy analysis. One is a hydrophobic region, which presumably contains six transmembrane segments. The other is a hydrophilic domain, which shows striking similarity to the sequences of the ATP-binding cassette (ABC) transporter proteins, including bacterial periplasmic transport proteins, the human multidrug resistance P-glycoprotein (MDR1), cystic fibrosis transmembrane conductance regulator (CFTR), and the putative adrenoleukodystrophy gene product (ALDP). Based on its transmembrane structure and the homology to ABC proteins, PMP70 may be involved in ATP dependent transport through peroxisomal membrane. PMID- 8411713 TI - [Molecular genetics of adrenoleukodystrophy]. AB - Human Xq28 region harbors many disease genes including genes for adrenoleukodystrophy, Emery-Dreifuss muscular dystrophy, X-linked centronuclear myopathy, and nephrotic diabetes inspidus. The genes for the diseases, however, have not been identified. On the other hand, only small number of transcribed sequences including G6PD gene, Gdx, P3, factor VIII gene, red and green color pigment genes, GABRA3 gene, L1 adhesion molecule gene, QM gene and so on have been identified at Xq28. To identify the disease genes at Xq28 by positional cloning, it is essential to construct physical maps of the Xq28 region and to develop a strategy for identifying expressed genes. Macrorestriction maps of human Xq28 have been generated by pulsed field gel electrophoresis (PFGE). With the recent development of yeast artificial chromosomes (YACs), major efforts have been focused on the generation of contigs of YACs from Xq28. Recently, a putative ALD gene was identified. The gene named ALDP gene was partially deleted in 6 of 85 independent patients with ALD. In familial cases, the deletions segregated with the disease. The deduced protein sequence of ALDP shows significant sequence identity to a peroxisomal membrane protein of 70 K that is involved in peroxisome biogenesis and shares unexpected homology to ABC transporter gene. PMID- 8411714 TI - [Mitochondrial encephalomyopathies: pleomorphism of the mitochondrial DNA mutations and clinical features]. AB - Recent studies analyzing mtDNA have established to elucidate the molecular pathology of mitochondrial encephalomyopathies. The human mitochondrial genome is 16,569 bp circular double-stranded molecule that is maternally inherited. Since the first report on large deletions of mtDNA in patients with progressive external ophthalmoplegia (PEO) by Holt et al in 1988, various mtDNA mutations were found. On the basis of the recent findings of mtDNA mutations, genetic classification of mitochondrial diseases has been proposed by S DiMauro in 1991. (1) large deletions or duplications of mtDNA were found in PEO and Pearson disease. (2) A single base substitution were reported in several mitochondrial encephalomyopathies as follows: (a) At nucleotide position 11778, 4136 or 4160......Leber's hereditary optic neuritis, (b) 8344......MERRF, (c) 3243 or 3271......MELAS, (d) 8993......Holt's disease. PMID- 8411715 TI - [MELAS (mitochondrial myopathy, encephalopathy lactic acidosis, and stroke-like episodes): clinical features and mitochondrial DNA mutations]. AB - MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) is one of the clinically-defined mitochondrial diseases, characterized by early onset and stroke-like symptoms. A point mutation at nucleotide pair 3243 within the tRNA-Leu (UUR) gene is found in 80% of MELAS patients and another mutation at nucleotide pair 3271 in 10%. In vitro and in vivo expression studies on 3243 mutant genome show that it affects both the transfer RNA and transcription termination functionally. By virtue of further analyses on relationship between the mutations and phenotypes, a new approach to deal with the disease could be obtainable. PMID- 8411716 TI - [Molecular genetic analysis for myoclonus epilepsy associated with ragged-red fibers (MERRF)]. AB - Two mutations in tRNA(Lys) gene of mitochondrial DNA were detected as the causes of this disease. We reviewed our previous studies and the recent literatures. We analyzed the mtDNA nucleotide sequence of a MERRF patient, the original case of MERRF described by Fukuhara et al., and identified a point mutation of 8,344 in tRNA(Lys) gene. This mutation detected in all 8 MERRF patients from 6 independent families, and not detected in 15 controls by polymerase chain reaction using a mismatched primer. We also quantitated the degrees of heteroplasmy of the point mutation at nt 8,344 of tRNA(Lys) in various postmortem tissues from two patients with MERRF. The percentages of the mutant mtDNA were similar in both clinically affected and unaffected tissues. PMID- 8411717 TI - [Deletions of mitochondrial DNA in Kearns-Sayre syndrome]. AB - Single large-scale deletions of mitochondrial DNA (mtDNA) are found in 70 to 80% of Kearns-Sayre syndrome (KSS) patients. Most deletions are flanked by direct repeats up to 13 nucleotides in length. The incidence of ragged-red fibers and cytochrome c oxidase-negative fibers in muscle is correlate with the amount of deleted mtDNA. Recently, study with 'cybrid' cell lines, which have different proportions of deleted mtDNA, showed that accumulation of deleted mtDNA to over 60% of the total mtDNA resulted in progressive inhibition of overall mitochondrial translation, as well as, reduction of cytochrome c oxidase activity. These results suggest that deletion of mtDNA alone is sufficient for the mitochondrial dysfunction in KSS. PMID- 8411718 TI - [Multiple mitochondrial DNA deletions in chronic progressive external ophthalmoplegia (CPEO)]. AB - We reviewed familial cases of chronic progressive external ophthalmoplegia (CPEO) associated with multiple mitochondrial DNA (mtDNA) deletions. A new case of familial CPEO with multiple mtDNA deletions, which were detected in the proband's skeletal muscles by Southern blotting and in all the tissues examined by using the polymerase chain reaction is also described. There was an approximate correlation between the clinical severity of the muscle involvement and the amount of mtDNA with deletions. Most of these familial CPEO cases, with multiple mtDNA deletions, exhibited an autosomal dominant mode of transmission. Abnormalities of some nucleus-driven factors, involved in the replication of mtDNA, may result in these multiple mtDNA deletions. PMID- 8411719 TI - [Molecular genetic analysis for Leber's hereditary optic neuropathy (LHON)]. AB - Leber's hereditary optic neuropathy (LHON) is a maternally inherited disease characterized of acute visual loss predominantly affecting young men. So far twelve mutations in the mitochondrial DNA have been associated in this disease. We reviewed these mutations, and showed methods to detect these mutations which are being used in our laboratory. Seven mutations (nps 3460, 5244, 7444, 11778, 13708, 15257, 15812) can be detected by the the loss of the sites, and 4 mutations (nps 4216, 4917, 11778, 3394) can be detected by the appearance of the restriction sites. Since only two mutations (nps 4160 and 14484) don't affect the restriction sites, we use mismatched primers as a method to detect the mutations by polymerase chain reaction. PMID- 8411720 TI - [Leigh's syndrome and mitochondrial myopathy]. AB - Leigh's syndrome is a subacute encephalopathy with characteristic pathological features and lactic acidosis. This syndrome is due to the disturbance of aerobic metabolism. Pyruvate dehydrogenase deficiency and cytochrome c oxidase deficiency are common metabolic disturbances in this syndrome. Complex I or II deficiency has also been claimed. Recently, mutations of mitochondrial genome have been also identified in some cases with Leigh's syndrome: np 3243 T to C, np 8344 G to A and np 8993 T to G. The possible correlation between phenotype and genotype in this heterogeneous syndrome was discussed. PMID- 8411721 TI - [The GABAA receptor beta 3-subunit gene (GABRB3) as a candidate responsible for central nerve disturbances in Angelman syndrome (AS)]. AB - AS is characterized by severe mental retardation, seizures, ataxic gait and easily evoked laughter. About 70 approximately 80% of AS patients have a chromosomal/molecular deletion at 15 q11-q13, occurring exclusively in the maternally-derived chromosome 15. There have been 4 AS patients whose chromosomes 15 are paternal uniparental disomy. This biased parent-of-origin suggests that genomic imprinting may play a role in the occurrence of the syndrome. GABRB3 is located at 15 q11-q13. GABAA is a main inhibitory neurotransmitter in the central nervous system (CNS) and functions through its receptor. The beta 3 subunit, one of the components of the receptor, is present in the telephalonal cortex, hippocampus, thalamus and cerebellum, and a peak GABRB3 expression is observed during embryogenesis. This indicates that GABRB3 plays a role in CNS development, suppression of seizures and behavioral control. Since GABRB3 is encompassed within the smallest deletion among AS patients, it becomes a candidate responsible for the central nerve disturbances in AS. This smallest deletion was found in 3 AS sibs, their phenotypically normal mother and maternal grandfather in a family, suggesting that the paternally-derived deletion has no phenotypical effect in the offspring but the maternally-derived one. However, recent studies demonstrated that the mouse Gabrb3 is not involved in imprinting. The confirmation of GABRB3 to be the AS gene needs to provide direct evidence of its imprinting. Our preliminary study showed that GABRB3 was not expressed in hydatidiform mole that is composed only of the paternal genome, while it was expressed fully in normal villous tissue, suggesting that GABRB3 is paternally imprinted.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411722 TI - [A novel primer extension method to detect the number of CAG repeats in the androgen receptor gene in families with X-linked spinal and bulbar muscular atrophy]. AB - X-linked spinal and bulbar muscular atrophy (SBMA), an adult-onset form of motor neuron disease, was recently reported to be caused by amplification of the CAG repeats in the androgen receptor gene. We report here a simple and rapid strategy to detect the precise number of the CAGs. After the DNA fragment containing the CAG repeats is amplified by the polymerase chain reaction, a primer extension is carried out; the extension of the end-labelled reverse primer adjacent to 3' end of CAG repeats stops at the first T after CAG repeats with the incorporation of dideoxy ATP in the reaction mixture. The resultant primer products are analysed by denaturing polyacrylamide gel electrophoresis and autoradiography. This method could be quite useful to detect not only CAG repeats in SBMA but also other polymorphic dinucleotide and trinucleotide repeats. PMID- 8411723 TI - [Molecular genetics of Duchenne/Becker muscular dystrophy]. AB - The X-linked gene responsible for Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) encodes dystrophin, a high-molecular-weight cytoskeletal protein. The identification of the dystrophin gene through positional cloning, and the subsequent description of its protein product have opened several new fields of research and genetic diagnosis. Studies in our laboratory revealed that 26 out of 47 (55%) cases of DMD and nine out of 12 (75%) cases of BMD exhibited genomic deletion. The DMD phenotype is associated with mutations that shift the reading frame of the message, whereas the BMD phenotype is associated with mutations that maintain the reading frame. Immunofluorescence microscopy has established dystrophin's distribution on the plasma membrane of muscles. DMD patients demonstrate a lack of dystrophin on their muscle cell membrane, whereas BMD patients produce a limited amount of protein or abnormally sized protein. Extensive studies on dystrophin and the gene may lead to an understanding of the cause for this and may allow development of a rational treatment for DMD to be developed. PMID- 8411724 TI - [Clinical and genetic heterogeneity in familial amyloidotic polyneuropathy associated with variant transthyretin]. AB - Familial amyloid polyneuropathy (FAP) is an autosomal disease, usually associated with a variant of transthyretin (TTR). To date, about 30 variants of TTR have been described in FAP. Clinical heterogeneity regarding age of onset and organ involvement exists in TTR-related FAP. The age of onset in patients with TTR-Met 30 variant ranges from the third to the seventh decade. The pattern of peripheral neuropathy varies considerably. Lower limb neuropathy is the common mode in cases with Met 30 and other many variants of TTR, while in some cases, upper limb neuropathy with carpal tunnel syndrome is the most important feature. The presence of vitreous opacity is one of the cardinal manifestations in several kindreds (eg. Cys 114, lle 33). The mechanism by which variant TTR affects clinical heterogeneity in FAP is unknown and deserves future study. PMID- 8411725 TI - [Prenatal diagnosis of familial amyloidotic polyneuropathy]. AB - Utilization of the in vitro DNA amplification method for prenatal diagnosis of familial amyloidotic polyneuropathy (FAP) in a German family was reported at the 1st symposium on FAP and other transthyretin related disorders, which was held in Granja, Portugal, in September 1989. Fetal DNA was prepared from chorionic villi and a part of the transthyretin (TTR) gene that may carry the FAP type I mutation was enzymatically amplified. The presence of the mutant TTR gene in fetal DNA was demonstrated by restriction analysis of the amplified DNA. Two other groups also reported prenatal diagnosis of FAP by polymerase chain reaction and restriction analysis. PMID- 8411726 TI - [Mutation analysis of amyloid precursor protein in early-onset familial Alzheimer's disease]. AB - Recently, three different mutations have been found at codon 717 of the amyloid precursor protein (APP) gene, changing the native valine to isoleucine, phenylalanine and glycine in some familial Alzheimer's disease (FAD) kindreds. More recently, some other mutations have also been reported at codons 670 and 671 (double mutation), and codon 692. As analysis of coding region except exon 16 and 17 has been insufficient in previous reports, we analyzed entire coding region of the APP gene of 6 Japanese early-onset FAD kindreds using automated sequencer. Three FAD families showed known 717 Val to Ile mutation, whereas no novel mutations were detected. PMID- 8411727 TI - [Absence of linkage disequilibrium at amyloid precursor protein gene locus in Japanese familial Alzheimer's disease with 717Val-->Ile mutation]. AB - To date, eleven independent early onset familial Alzheimer's disease (EOFAD) pedigrees with the 717Val-->Ile mutation of amyloid precursor protein (APP) gene have been identified. Interestingly, five pedigrees have been of Japanese origin. The apparent ethnic prediction of this mutation raises the possibility that there is a founder effect in Japan. If the hypothesis holds true, we can expect the presence of linkage disequilibrium at the APP locus. We did not, however, observe any significant linkage disequilibrium at any locus of APP or the adjacent GT12 locus in the five Japanese EOFAD probands with the 717Val-->Ile mutation. The result indicates that a founder effect would probably not be present in Japanese EOFAD pedigrees with the 717Val-->Ile mutation. PMID- 8411728 TI - [Molecular genetics of neurofibromatosis type 1]. AB - The gene responsible for neurofibromatosis type 1 (NF1) encodes a protein similar to GTPase-activating protein (GAP) for the products of ras protooncogenes. Since NF1 protein possesses GAP-like activity, it is suggested that NF1 protein plays a role in the regulation of ras-mediated signal transduction. Impaired function of NF1 protein could therefore, lead to abnormal growth or differentiation of neural crest-derived cells in NF1 patients. Mutations of NF1 gene are also found in sporadic tumors not associated with NF1, supporting the hypothesis that NF1 gene acts as a tumor suppressor gene. PMID- 8411729 TI - [Molecular biological analysis of neurofibromatosis type 2 gene]. AB - Neurofibromatosis type 2 is a hereditary disorder characterized by bilateral vestibular and other schwannomas as well as various other central nervous system neoplasms such as gliomas, meningiomas, and neurofibromas. The region containing the NF2 gene has been localized to 22q12 both by linkage analysis and deletion mapping of NF2-related tumors. Recently a candidate gene, named Merlin, was cloned by means of defining a constitutional interstitial deletion in an NF2 patient. The Merlin gene product belongs to a family of proteins which include moesin, ezrin and radixin, involved in linking the cytoskeleton to the cell membrane. Merlin thus forms a novel class of tumor suppressor genes, and an analysis of its function in vivo and the clinical implications of its loss in schwannoma cells remains to be explored. PMID- 8411730 TI - [Recent progress of research on hereditary spinocerebellar degeneration]. AB - Hereditary spinocerebellar degeneration consist of a cluster of heterogenous disorders. Based on clinical features, neuropathological findings and inheritance mode, these disorders have been classified into several entities. In the Japanese, olivo-ponto-cerebellar atrophy (OPCA), Machado-Joseph disease (MJD) and dominant cortical cerebellar atrophy (CCA) are the most popular forms of ataxia. Recent linkage analyses have revealed the locations of these genes; OPCA maps either on 6p23 (SCA1) or on 12q23 (SCA2), and MJD on 14q. The chromosomal assignment of these genes has resulted in development of a new classification of ataxia and has opened a new era of ataxia research. PMID- 8411731 TI - [Advances in molecular genetics of myotonic dystrophy]. AB - Myotonic dystrophy (DM) is the most common muscular dystrophy affecting adults among Caucasian and Japanese populations, with an average incidence of 12.5 in 100,000 in Caucasians and 5.5 in 100,000 in the Japanese. Recently the DM gene was cloned and characterized showing homology with the family of serine-threonine protein kinase. In DM patients expansion of an unstable trinucleotide CTG repeat, located within the 3' untranslated region of DM kinase gene, is involved. In this review we outline the molecular biological aspects of DM including DNA diagnosis, anticipation, differences between paternal and maternal transmission, founder chromosome and expression of the gene. PMID- 8411732 TI - [Huntington's disease--advances in gene mapping]. AB - Huntington's disease (HD) is a progressive neurodegenerative disorder characterized by motor disturbance, cognitive loss, and psychiatric manifestations. It is inherited in an autosomal dominant fashion. The genetic defect causing HD was assigned to chromosome 4 in 1983 using polymorphic DNA markers in humans. Thereafter, a location cloning approach was pursued to isolate and characterize the HD gene. Recently, the Huntington's disease collaborative research group has isolated a new gene, IT 15, in 4p 16.3. IT 15 contains a polymorphic trinucleotide repeat that is expanded and unstable on HD chromosomes. A (CAG)n repeat longer than the normal range was observed on HD chromosomes from disease families. The (CAG)n repeat appears to be located within the coding sequence of a predicted 348 kd protein that is unrelated to any known gene. PMID- 8411733 TI - [A trend of molecular genetics on prion diseases and prion protein]. AB - Infectious amyloid filaments designated as prion rods or scrapie associated fibrils (SAF) present in brain tissues affected by transmissible spongiform encephalopathies such as Creutzfeldt-Jakob disease (CJD), Gerstmann-Straussler Scheinker disease (GSS) and kuru of humans, and scrapie of sheep. A hydrophobic glycoprotein, PrPSc is a major component of SAF, and is known to be associated with the infectivity of these diseases. Both PrPSc and the normal isoform of this glycoprotein, PrPC are encoded by a single host gene, PrP gene, and the conversion of PrPC to PrPSc is a posttranslational event. Several mutations on the PrP gene are associated with variations of the phenotype and the occurrence in familial CJD and GSS. PMID- 8411734 TI - Excess incidence of lung cancer among pulmonary tuberculosis patients. AB - Reviewing the epidemiological studies on coexistent pulmonary tuberculosis and lung cancer since 1960, it was confirmed that patients with active pulmonary tuberculosis have a higher risk of dying from lung cancer or other malignancies, in spite of a very high mortality from tuberculosis per se. Females showed a higher risk than males. An antagonistic hypothesis between the above two diseases since 1854 seemed based on the facts that tuberculosis patients had mostly died in young age groups and had had little chance of surviving to cancer age groups before the advent of modern treatments, and that the lack of an adequate disease registration system might have caused a failure to reveal any association between the two diseases. Specific causative factors of the excess incidence of cancer among active pulmonary tuberculosis patients, however, have so far not be clarified. PMID- 8411735 TI - Human papillomavirus DNA in squamous cell carcinomas of the respiratory and upper digestive tracts. AB - The prevalence of human papillomavirus (HPV) types 16 and 18 in clinical samples of squamous cell carcinomas from respiratory and upper digestive tracts was studied. HPV DNA of types 16 and/or 18 was detected using the polymerase chain reaction (PCR) method in 16 out of 121 cases (13.2%). By Southern blot hybridization, however, only the DNA from a laryngeal and a tonsillar carcinoma was found to hybridize with the whole HPV 16 DNA probe (two out of 16 HPV DNA positive cases by PCR, 12.5%). None of the DNAs hybridized with the whole HPV 18 DNA probe. The discrepancy in the results of PCR and Southern blot hybridization methods seemed to reflect their sensitivity. The possible relation between prevalence of HPV DNA and carcinogenesis in respiratory and upper digestive tract is discussed. PMID- 8411736 TI - Mechanism of the discrepant effect of a combination of methotrexate plus dipyridamole on human hematologic cell lines. AB - Mechanisms of the discrepant effect of methotrexate and dipyridamole on human hematologic cultured cell lines were investigated by analyzing intracellular methotrexate levels and thymidine incorporation through the salvage pathway, since the combination of methotrexate and dipyridamole has different effects according to cell type: additive effects on ML-1 and THP-1 (myelo-monocytoid cells); reduced effects on MOLT-3, SKW-3, P32/ish and BL-TH (lymphoid cells). Dipyridamole reduced the toxicity of methotrexate by diminishing intracellular methotrexate levels in MOLT-3 and BL-TH (lymphoid cells), in which the reduction of intracellular methotrexate affected more than just the blocking of the salvage pathway required for growth by dipyridamole. On the other hand, dipyridamole enhanced the toxicity of the combination by blocking the salvage pathway in an ML 1 (myelo-monocytoid cell) and in a methotrexate-resistant subline of BL-TH/MTX (lymphoid cell), in which the salvage pathways were considered activated. Dipyridamole could prove to be a useful drug for reversing the drug resistance caused by the activation of the salvage pathway. PMID- 8411737 TI - Organ function index: a predictive marker of operative mortality based on multiple organ dysfunctions in patients with esophageal cancer. AB - To evaluate the risk of transthoracic esophagectomy for cancer based on the overall impairment of vital organs, we examined pulmonary, cardiac, renal and hepatic functions using 18 markers in 35 patients between 1982 and 1984. A discriminant analysis proved useful in determining whether or not the patients would be at risk of operative mortality (operative death and hospital death), based on the overall impairment of vital organs. The accuracy of the prediction of operative mortality by this model, the organ function index (OFI), was 91.4%. The OFI was then applied to 66 patients seen between 1986 and 1991. During this period, a change in policy for performing transthoracic esophagectomy and perioperative care on patients with impairment of multiple organs was associated with a decrease in operative mortality. We conclude the OFI to be beneficial in evaluating the risk of operative mortality based on mild to moderate dysfunctions of multiple vital organs in patients with esophageal cancer. PMID- 8411738 TI - MX2; 3'-deamino-3'-morpholino-13-deoxy-10-hydroxycarminomycin (KRN8602) in refractory metastatic breast cancer: results of a preliminary phase II trial. AB - We performed a preliminary phase II clinical trial of MX2; 3'-deamino-3' morpholino-13-deoxy-10-hydroxycarminomycin (KRN8602) in patients with metastatic breast cancer who had failed to respond to previous chemotherapeutic regimens after clinical evidence of systemic disease. Twelve patients at a single institute received KRN8602 at a dose of 35 mg/m2 intravenously once every three weeks. All the patients were followed-up until their disease progressed. There was one complete response lasting 17 weeks and one partial response lasting eight weeks. Among the 12 patients, World Health Organization (WHO) grades 3 and 4 neutropenia were observed in five and two patients, respectively. Grade 3 anemia was observed in three patients but severe thrombocytopenia was not observed. Grade 3 nausea/vomiting was observed in eight patients. Alopecia was not observed. The results of this preliminary phase II trial suggest a need for further testing of the anti-tumor activity of KRN8602 in patients with metastatic breast cancer. PMID- 8411739 TI - Toxicity grading criteria of the Japan Clinical Oncology Group. The Clinical Trial Review Committee of the Japan Clinical Oncology Group. PMID- 8411741 TI - [Autoimmune hepatitis]. PMID- 8411740 TI - Combination chemotherapy for a senile patient with adenoid cystic carcinoma of the esophagus: a case report. AB - Adenoid cystic carcinoma of the esophagus with multiple liver metastases was found in a 77-year-old Japanese female in October, 1991. Combination chemotherapy, including etoposide (VP-16) and tegafur (FT) allowed the patient to survive for approximately a year after the onset of the initial symptoms. During her course of treatment, the patient improved greatly with minimal toxicity, and declines in the elevated levels of GOT and LDH were also noted. Since no other therapy, such as surgery or radiation, was given to the patient, the response to chemotherapy as well as the survival advantage for the patient have been evaluated. PMID- 8411742 TI - [Propranolol for prophylaxis of first hemorrhage in cirrhotic patients with esophageal varices--a controlled study comparing with sclerotherapy]. AB - To compare the efficacy of oral propranolol and sclerotherapy in the prevention of first hemorrhage from esophageal varices, 65 cirrhotic patients with moderate to large esophageal varices and no history of bleeding were included in the prospective controlled trial. After randomization, 33 patients received propranolol at a does reducing the heart rate by 25%; 32 patients were treated with intra-variceal and extra-variceal injection of ethanolamine oleate. On entry to the trial, the two groups were comparable in terms of clinical and biological parameters. The patients were observed for up to 60 months, with an average of 31 months. Nine patients bled (5 in propranolol and 4 in sclerotherapy) during follow-up. No significant difference were observed between propranolol and sclerotherapy in the cumulative bleeding rate and survival. The multivariate Cox model indicated that drug compliance in the propranolol group and high portal pressure in the sclerotherapy group were factors predictive of the first hemorrhage. These data support that propranolol and sclerotherapy are of comparable value in preventing the first hemorrhage in cirrhotic patients with esophageal varices. PMID- 8411743 TI - [Clinical relevance of abdominal imaging examinations in malignant lymphoma]. AB - We prospectively evaluated the clinical importance of abdominal imaging examinations (US, CT, upper gastrointestinal barium X-ray) in 233 consecutive patients who gave informed consent for the examinations. The examinations revealed intra-abdominal lesions in 99 of 233 patients. Intra-abdominal lymph nodes were most frequently affected, followed by stomach, spleen, liver, small intestine and large intestine. In Hodgkin's lymphoma, no gastrointestinal involvement was noted but one in the small intestine. Prognosis was poorer with advancing stage according to Ann Arbor classification. However, the presence or absence of intra-abdominal lesions did not influence the prognosis when patients were matched for the stage. Abdominal imaging examinations altered the bed-side staging to more advanced stages in 22 of 163 patients with stage I through stage III lymphoma, influencing prognosis as well as the decision of therapeutic modalities. These three diagnostic modalities were complementary to one another. In conclusion, every one of these abdominal imaging examinations is important for planning the management of patients with malignant lymphoma. PMID- 8411744 TI - [Clinico-pathological studies on the lymph node metastases of the gastric cancer invading to proper muscle layer]. AB - In 65 patients who underwent surgical resection of the gastric cancer invading to proper muscle layer (pm cancer), lymph node involvement at the time of resection and the site of recurrence were investigated. Patients who underwent gastrectomy for submucosal (sm cancer) or subserosal (ss alpha.beta cancer) gastric cancers during the same period served as controls. Lymph node involvement was positive at a higher rate in the pm cancer than in the sm cancer, but the rate for number of lymph node metastases was not dependent on the stage of lesions. Of the lesions in the pm cancer, those of the Borrmann type involved lymph nodes at a particularly high rate. Of the metastatic pattern in the lymph nodes, the rate of severe metastases in pm cancer was higher than in the sm cancer. In the pm cancer, the cancer recurred in the liver and peritoneum, but not in any lymph nodes, thus suggesting that the surgical resection and the dissection of regional lymph nodes had been successful. PMID- 8411745 TI - [Availability of determination of serum anti-Helicobacter pylori IgG antibody in diagnosis of Helicobacter pylori infection]. AB - We studied on clinical availability of determination of serum anti-Helicobacter pylori (HP) IgG antibody in diagnosis of HP infection compared with bacterial culture. The population of this study consisted of patients who underwent endoscopic examination in our hospital. Detection and quantification of HP were made by the culture of biopsy specimens taken from the antrum and the body of the stomach. And, simultaneously, serum anti-HP antibody (HP-Ab) was measured by ELISA. The levels of HP-Ab in culture positive patients, 40.6 +/- 33.7U, are significantly higher than that of culture negative patients, 10.6 +/- 9.1U. Sensitivity and specificity in diagnosis of HP infection using determination of serum HP-Ab were 96.2% and 60.0% respectively. Significant correlation was found between the levels of serum HP-Ab and the amounts of HP bacilli in the biopsy specimen taken from body of the stomach. PMID- 8411746 TI - [Relationship between biliary excretion of conjugated sulfobromophthalein and glutathione disulfide in rats]. AB - Relationship between biliary excretion of sulfobromophthalein conjugated with glutathione (BSP-GSH) and glutathione disulfide (GSSG) was investigated in Sprague-Dawley rats. BSP-GSH solution was infused intravenously at three different rates. After administration of a glutathione-oxidizing agent, diamide, biliary excretion of GSSG increased temporarily and that of BSP-GSH decreased during the same period. A linear correlation was found between the increments in biliary excretion of GSSG and the decrements in that of BSP-GSH only when BSP-GSH was infused at a rate near its biliary transport maximum. The results may indicate competition in vivo between GSSG and BSP-GSH for a common transporter on canalicular membrane. PMID- 8411747 TI - [The effects of Ara-A on viral markers in duck hepatitis B virus carrier ducks]. AB - Duck hepatitis B virus (DHBV) carrier ducks of one week old were injected with Ara-A (adenine arabinoside) of different dose including 2.5 (11 ducks), 5.0 (11), 10.0 (10) and 20.0 (10) mg/kg for 14 days. This antiviral effect showed dose dependence up to 5.0 mg/kg and this dose seemed effective to obtain significant antiviral effect. Viral DNA and DNA polymerase activity were reduced significantly from the 1st week after starting the administration of Ara-A. This antiviral effect was maintained even at the 1st week after discontinuation of the drug. These findings were quite similar to those observed in HBV carriers. With the increasing necessity of Ara-A treatment in patients who will not respond to interferon therapy, DHBV seemed a suitable model for the investigation of the dose and antiviral effect of Ara-A treatment in humans. PMID- 8411748 TI - [A case report of a huge exogastric, pedunculated gastric leiomyoma]. PMID- 8411749 TI - [A case of ascending colon cancer with a tumor thrombus in the superior mesenteric vein]. PMID- 8411750 TI - [Acute hemorrhagic rectal ulcer with many times of massive rectal bleeding: report of a case--including study of vessel reconstruction on the resected rectum]. PMID- 8411751 TI - [A case of liver cirrhosis with portal vein calcification associated with hypoglycemia induced by branched chain amino acid solution]. PMID- 8411752 TI - [A case report of endocrine cell carcinoma of the liver]. PMID- 8411753 TI - [A case of chronic hepatitis C with pneumonitis during interferon therapy]. PMID- 8411754 TI - [An autopsied case of primary hepatic malignant lymphoma associated with chronic active hepatitis C]. PMID- 8411755 TI - [A case of primary biliary cirrhosis associated with chronic active hepatitis type C improved by the injection of interferon and the administration of ursodeoxycholic acid]. PMID- 8411756 TI - [A case of Laurence-Moon-Biedl syndrome accompanied by congenital common bile duct dilatation and common bile duct carcinoma]. PMID- 8411757 TI - [A case of small cell carcinoma of the pancreas, response to the modified CHOP therapy]. PMID- 8411758 TI - [Thrombospondin modulates cell attachment, spreading and proliferation in cultured rat mesangial cells]. AB - The effect of thrombospondin (TSP) on attachment, spreading, cell toxicity and proliferation of cultured rat mesangial cells (M cell) were investigated. The results are as follows: 1. Attachment and spreading of M cell on the culture plates coated with TSP were inhibited in a dose dependent manner, 2. 50 micrograms/ml TSP in the medium brought about the significant detachment of the adhered M cell. 3. TSP had no demonstrable effects on 51Cr-release from M cell, 4. 3H-thymidine uptake into M cell was inhibited by TSP in a dose dependent manner. These results indicate that TSP modulates attachment, spreading and proliferation of M cell. Therefore, it is suggested that overproduction of TSP in M cell under some pathophysiologic conditions such as diabetic nephropathy may influence the M cell functions in a self-suppression. PMID- 8411759 TI - [Serum superoxide dismutase (SOD) activity in patients with renal disease by a spintrap method using electron spin resonance (ESR)]. AB - Superoxide dismutase (SOD) activity in serum samples of patients with chronic glomerulonephritis (CGN) and chronic renal failure (CRF) was measured by a spin trap method using electron spin resonance (ESR). Twenty-three patients with CGN, 10 patients with CRF and 10 healthy adults were examined. Among 23 patients with CGN, there were 12 patients with IgA nephropathy and one patient with membranous nephropathy diagnosed by immunofluorescence of renal biopsy specimens. Other CGN patients were diagnosed by its clinical criteria. The serum activity of SOD in patients with CGN or CRF was significantly higher than those in healthy adults (p < 0.05). The serum SOD activity in patients with CRF was also higher than those in patients with CGN (p < 0.05). Marked high levels of serum SOD activity were observed histologically in the advanced stage of IgA nephropathy. These results suggest an increase in serum SOD activity may reflect renal injuries in patients with CGN and CRF. PMID- 8411760 TI - [Clinical significance of microalbuminuria in patients with rheumatoid arthritis]. AB - In order to make an estimate of clinical significance of microalbuminuria (MAU) in patients with rheumatoid arthritis (RA), we studied MAU in 138 patients with RA without macroalbuminuria. MAU was assayed by double-antibody RIA in the ambulatory urine. Moreover, urinary (U) beta 2-microglobulin (BMG) and N-acetyl beta-D-glucosaminidase (NAG) were simultaneously measured. The values for MAU/U creatinine (U-Alb index) in patients with RA, osteoarthropathy (OA) and normal controls were 25.7 +/- 38.2, 11.4 +/- 11.5 and 7.7 +/- 3.5, respectively, and U Alb indices in patients with RA were significantly higher than U-Alb indices in patients with OA and normal controls. Especially, in patients with RA receiving lovenzarit disodium and gold sodium thiomalate (GST), U-Alb indices were elevated. U-Alb indices were not correlated with clinical findings in RA. Also, U Alb indices were not correlated with U-BMG indices and U-NAG indices in patients with RA. In serial measurements of U-Alb index, U-BMG index and U-NAG index in a patient with RA who developed massive macroalbuminuria during GST therapy, it was found that U-Alb index was elevated first, followed by U-NAG index and finally U BMG index was elevated. These results indicate that U-Alb indices are elevated in patients with RA without macroalbuminuria, and serial measurements of MAU in patients with RA, especially receiving disease modifying antirheumatic drugs, are useful for the detection of subclinical glomerular injury. PMID- 8411761 TI - [Lp(a) lipoprotein in patients on maintenance hemodialysis--a study from apo(a) isoform]. AB - It has been suggested that lipid abnormalities may be involved in the development of cardiovascular disorders in patients on maintenance hemodialysis (HD). Hypertriglyceridemia commonly accompanies this condition and is associated with decreased concentrations of HDL-cholesterol. Recent clinical interest has been paid to the disturbances of Lp(a) and apo (a) isoform in relation to cardiovascular disorders. Although high concentrations of Lp(a) are associated with ischemic heart disease, we are unaware of the availability of such data concerning patients on maintenance hemodialysis. We therefore compared levels of Lp(a) and the frequency of occurrence of their isoforms in 310 patients (chronic glomerulonephritis, N = 221; diabetic nephropathy, N = 77; polycystic kidney disease, N = 12) on maintenance hemodialysis and in 212 normal subjects. The following results were obtained. (1) HD patients showed significantly elevated levels of Lp(a) compared to normal subjects. (2) Studies of apo (a) isoform showed that HD patients showed high frequency of S2 and low frequency of S4. (3) HD patients, especially long-term patients, showed high frequency of double band (S2/S3). (4) There were no significant differences in the levels of Lp(a) and in the frequency of apo(a) isoform among 3 different etiological studies. PMID- 8411762 TI - [A study on uric acid excretion in patients with hypouricemia]. AB - We studied four patients with hypouricemia and almost normal renal function. Two patients are idiopathic renal and familial. The other two cases are secondary to diabetes mellitus. The study of the uricosuric mechanism with pyrazinamide (PZA) and benzbromarone (Bb) test shows that two patients have a defect of pre secretory reabsorption of urate, the other one a incomplete combined defect of pre-and post-secretory reabsorption, and the other one a enhanced secretion of urate. The fact that two patients have the history of acute renal failure (ARF) suggested that strenuous exercise or dehydration would be responsible for ARF in patients with renal hypouricemia. PMID- 8411763 TI - [Mechanisms of prevention of progression of diabetic nephropathy by dietary protein restriction]. AB - This study was designed to clarify the mechanisms by which the progression of diabetic nephropathy is inhibited by low-protein diet. Six control and twenty five streptozotocin-induced diabetic rats were divided into 4 group: Group I consisted of non-diabetic control rats administered a 40% protein diet (n = 6); Group II, diabetic rats administered a 40% protein diet (n = 8); Group III, diabetic rats treated with a 10% protein diet (n = 8), and Group IV, diabetic rats treated together with a 10% protein diet and ad lib, drinking water containing 0.75 mg/l of angiotensin-converting enzyme inhibitor (ACEI) (n = 9). The follow-up period was 24 weeks. Urinary albumin excretion rate (AER), glomerular filtration rate (GFR) and morphometric measurement of renal biopsy specimens obtained at 12 and 24 weeks were evaluated. AER was significantly increased in the untreated diabetic Group II compared with the control Group I. However, AER was significantly decreased in the treated Group III and IV compared with the untreated diabetic Group II. There was no significant difference in AER between Group III and IV. Compared with Group II, the increase in GFR was significantly inhibited in Group IV but not in Group III. The increase in mesangial matrix and GBM thickening were both prominent in the untreated diabetic Group II, and significantly inhibited in the treated Group III and IV.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411764 TI - [Nutritional assessment of patients on continuous ambulatory peritoneal dialysis]. AB - Nutritional status was assessed in 214 CAPD patients from eight centers in Japan. All patients were receiving adequate dialysis, as indicated by KT/Vurea and Karnofsky's activity scale. Subjective global assessment indicated that 25% of 83 female patients and 27% of 131 male patients were, at least, moderately malnourished. Mid-arm muscle circumference was lower in malnourished male patients than in well-nourished patients, however, in female patients none of the anthropometric variables reflected nutritional status. Lower serum albumin level was observed in malnourished male patients, but not in female patients. Daily protein intake, estimated from protein catabolic rate (PCR) was approximately 0.8 g/Kg BW in both males and females. This value is similar to those reported in literatures for CAPD patients. In our well-rehabilitated patients, PCR did not reflect the nutritional status as indicated by subjective global assessment. The data suggest that low protein intake was not always the major cause of malnutrition in these patients and that other factors, such as endocrine disorder, may also have contributed to malnutrition. PMID- 8411765 TI - [A study on plasma allantoin in patients undergoing maintenance hemodialysis]. AB - Current studies have reported that free radical molecules had anticipated intimately with the pathogenesis of some complications specific for a long term hemodialysis. Among substances generated by free radical reaction, methylguanidine (MG) had been well known as most popular parameter in hemodialyzed patient, but allantoin (Alt) had been reported to be generated from uric acid by hydroxyradical, too. In this study, we measured serum level of Alt in long term hemodialyzed patient and analysed in the comparison with serum level of sialic acid, MG, and MG/Cr value. Serum level of Alt in patients group varied from 2.6 to 105.9 nmol/ml, which was significantly higher as compared with exclusively undetectable level in normal volunteers (n = 15). Serum level of Alt in each patient showed a significant correlation with serum level of a sialic acid (r = 0.400 p < 0.02). Furthermore, although serum level of Alt failed to show a significant correlation with serum level of MG, but showed a significant correlation with MG/Cr value. (r = 0.485 p < 0.01). In addition, higher level could be found in patient group having higher serum level of MG or MG/Cr value. Based upon those results, we proposed that a measurement of serum level of Alt could be of value in detection of free radical reaction in long term hemodialyzed patient. PMID- 8411766 TI - [Growth and serial changes of bone calcium content after renal transplantation in childhood]. AB - We evaluated metacarpal index (MCI) and bone mineral content (BMC) of right 2nd metacarpal bone X-ray films using the microdensitometer technique in 12 pediatric and 32 adult renal transplant (Tx) recipients. Grafts were well functioning for more than 1 year in all adults (serum creatinine < or = 2.0 mg/dl) and in 9 of the children (serum creatinine < or = 1.2 mg/dl). Immunosuppression consisted of cyclosporin (CyA), methylprednisolone (MPL), mizoribine and anti-lymphocyte globulin for all children. 18 of the adults were given CyA and 14 were given conventional immunosuppression. BMC was found to be increased in both children with good renal function and in adults. MCI was improved in 2 children with good renal function and in 2 adults using CyA. Immunosuppression of CyA and low dose MPL had an improving effect on renal osteodystrophy. Alternate-day MPL dosage was between 1, 6 and 7.3 mg/m2/day (mean 3, 6 mg/m2/day) in the 9 children with good renal function. Bone age of the children was seen to be developed in accordance with calendar age. Growth velocity of the 9 children with good renal function was better than the mean level of normal children. However, growth velocity at 3 years after Tx declined slightly, compared with that within 2 years. Similarly, somatomedin C was above the normal range within 2 years after Tx. Thus, bone metabolism after Tx may have been influenced by serial changes of somatomedin C. PMID- 8411767 TI - [A case of mesangioproliferative glomerulonephritis (GN) associated with unique lesions of juxtaglomerular apparatus (JGA) and interstitium]. AB - A case of mesangioproliferative glomerulonephritis (GN) associated with unique lesions of the juxtaglomerular apparatus (JGA) and interstitium is discussed. A 31-year-old Japanese woman who developed eyelid and pretibial edema with nephrotic-range proteinuria (4.8 g/day) and without hematuria, was admitted. Her proteinuria and edema quickly disappeared within 7 days after admission without treatment. Her blood examinations revealed hypocomplementemia on admission, but complement recovered to normal levels after 4 weeks. A renal biopsy specimen obtained on the 5th day of admission revealed moderate mesangioproliferative GN with marked periarteriolar inflammatory cell infiltrations in the JGA and occasionally in the tubular interstitium. Depositions of IgG, IgA, IgM and C3 were observed in the glomerular mesangial regions and some capillary walls, but not in the extraglomerular areas. Titers of GN-related viral antigens were not increased. Although the renal histology of this case was similar to that of experimental acute cytomegalovirus (CMV) GN in mice (described by Smith, R.D.), we could not detect CMV antigen by indirect immunofluorescent method or the virus like particles by electron microscopy. Clinical cases of nephropathy combining lesions of the glomerulus, JGA, and interstitium are very rare. We herein report a patient with mesangioproliferative GN, who underwent an acute clinical course associated with unique inflammatory lesions of the JGA and/or interstitium. PMID- 8411768 TI - [Focal glomerular sclerosis in a sibling]. AB - We described here one sibling with focal glomerular sclerosis. Proteinuria was noticed at the age of five in brother and four in sister. Both of them developed nephrotic syndrome shortly after the discovery of proteinuria. The nephrotic syndrome was resistant to corticosteroid, immunosuppressive agents or the combination of these drugs. Percutaneous renal biopsy in them revealed morphological and immunohistological features compatible to focal glomerular sclerosis. HLA typing in HLA-A, B, C and DR loci was identical to both. This observation suggests that genetic factors is associated with the pathogenesis of focal glomerular sclerosis. PMID- 8411769 TI - [A case of hyperammonemia in chronic renal failure successfully treated with the infusion of NaHCO3]. AB - The present report describes a rare of a 77-year-old woman who developed encephalopathy and metabolic acidosis associated with hyperammonemia, at the introduction of hemodialysis by chronic renal failure. With the intravenous infusion of HCO3-, levels of acidosis and hyperammonemia decreased rapidly. Concomitantly the disturbance of consciousness was improved. Results of plasma amino acid patterns of pre and post infusion of HCO3- showed improvement of the metabolism of the urea cycle, increased urea synthesis and decreased plasma ammonium levels. The role of the hepatic urea cycle has been considered to be exclusively the elimination of potentially toxic ammonia. In the conventional view, the acid base balance of the body obtains stabilized homeostasis by the function of the principal organs, lungs and kidneys. But, it has been recently shown that urea cycle is an important factor in the maintenance of pH homeostasis, due to regulated metabolism of HCO3-. Both HCO3- and NH4+ are converted to urea indicating the urea cycle's involvement in acid base homeostasis. 2HCO3- + 2NH4+-->urea+CO2+3H2O In this case, with the infusion of HCO3, the metabolism of the urea-cycle was improved and plasma ammonium levels were decreased. This indicates that HCO3- is an important factor for the metabolism of ammonia. PMID- 8411770 TI - Neuroprotective activity of fructose-1,6-bisphosphate following transient forebrain ischemia in the Mongolian gerbil. AB - We examined the protective activity of fructose-1,6-bisphosphate (FBP) on mortality and delayed hippocampal cell death induced by transient cerebral ischemia in the Mongolian gerbil. Forebrain ischemia was produced by bilaterally occluding the common carotid arteries for 15 min using microaneurysm clips; then the blood supply to the brain was restored. The number of survivors was counted for 8 days, and the histopathological damage in the CA1 region of the hippocampus was scored according to the semiquantitative scale of Rudolphi and colleagues. When injected 15 min before the ischemic insult, FBP (100 and 333 mg/kg, i.v.) significantly reduced the rate of mortality during the 8-day observation period. Equivalent doses of fructose and fructose monophosphate did not improve survival, and neither did low doses (33 mg/kg) of FBP. FBP also produced a significant degree of protection against the CA1 pyramidal cell loss in comparison with its vehicle (distilled water). Conversely, when we administered the compound, at the same dose, 15 min after the release of the arterial occlusion, we observed neither a significant reduction of mortality nor significant protection against hippocampal CA1 pyramidal cell loss. These results suggest that FBP possesses salutary properties against the damages induced by transient cerebral ischemia, although they are evident only when the compound is administered before the resolution of the ischemic injury. PMID- 8411771 TI - Effects of spiradoline mesylate, a selective kappa-opioid-receptor agonist, on the central dopamine system with relation to mouse locomotor activity and analgesia. AB - Neurochemical and behavioral investigations were made to assess the role of central dopaminergic systems in mouse locomotor activity and analgesia by spiradoline mesylate. Analgesic activities of the kappa-opioid-receptor agonists spiradoline and U-50488H were not altered by haloperidol or L-dopa, whereas morphine analgesia was enhanced by haloperidol but attenuated by L-dopa. Spiradoline decreased spontaneous locomotor activity in mice and inhibited methamphetamine- or morphine-induced locomotor activity. In contrast, morphine given alone increased locomotor activity and enhanced methamphetamine-induced locomotor activity. In a neurochemical study, spiradoline decreased the amounts of dopamine metabolites in the striatum, but did not alter them in the brainstem and cerebral cortex. Morphine increased the dopamine metabolite contents in all three brain regions tested. These results suggest that inhibition of the dopaminergic pathway in the brain by spiradoline may be involved in its suppression of locomotor activity, but not in its analgesia; whereas, stimulation of the dopaminergic pathway by morphine seems to function in both behaviors: enhancement of locomotor activity and inhibition of analgesia. PMID- 8411772 TI - Effects of NG-nitro-L-arginine on isolated rabbit afferent arterioles. AB - We examined the effects of NG-nitro-L-arginine (L-NNA) on isolated rabbit afferent arterioles to confirm that nitric oxide is released at the resistance vessel level in the kidney. We microdissected the superficial afferent arterioles from the kidneys of New Zealand White rabbits. Each afferent arteriole was cannulated with a micropipette system, and the intraluminal pressure was set at 80 mmHg. By our methods, we found that norepinephrine (NE) decreased the lumen diameter of the afferent arterioles in a dose-dependent manner, and acetylcholine increased the lumen diameter of NE-constricted afferent arterioles. L-NNA (10(-4) M) gradually decreased the lumen diameter of afferent arterioles from 21.5 +/- 0.9 to 18.6 +/- 0.9 microns in 20 min, but NG-nitro-D-arginine (10(-4) M) did not affect them (from 21.8 +/- 1.3 to 21.8 +/- 1.5 microns). L-Arginine (10(-2) M) restored the lumen diameter of L-NNA-contracted afferent arterioles to the control levels. These findings indicate that the isolated afferent arteriole has the ability to release or to synthesize and release nitric oxide under basal conditions and that this basal release of nitric oxide plays an important role in the basal tone of the afferent arteriole. PMID- 8411773 TI - In vitro pharmacology of a novel non-peptide angiotensin II-receptor antagonist, E4177. AB - E4177, 3-[(2'-carboxybiphenyl-4-yl)methyl]-2-cyclopropyl-7-methyl-3H- imidazo[4,5 b]pyridine, was characterized by in vitro autoradiography and by examining functional antagonism upon angiotensin II (Ang II)-induced contraction of isolated vessels. In rat adrenal cortex and liver, E4177 competitively inhibited the specific binding of 125I-[Sar1,Ile8]Ang II, with IC50 being (5.2 +/- 1.0) x 10(-8) M for the adrenal cortex and (1.2 +/- 0.3) x 10(-7) M for the liver. These IC50 values were similar to those for losartan, which showed an IC50 of (6.0 +/- 0.9) x 10(-8) M for the adrenal cortex and (1.3 +/- 0.5) x 10(-7) M for the liver. In contrast, E4177 and losartan had little effect on the binding to rat adrenal medulla where AT2-receptors predominate. These results indicate that E4177 is AT1-specific as is losartan. E4177 and losartan competitively antagonized the Ang II-induced contraction of human and rabbit arterial strips without any agonistic action. The obtained IC50 values indicated that E4177 was twice as potent as losartan in human arteries and three times more so in rabbit aortic strips. Responses to norepinephrine, serotonin, histamine or KCl were not affected by E4177. In addition, E4177 (10(-5) M) had no effect on angiotensin converting enzyme activity. These data indicate that E4177 is a potent AT1 Ang II receptor antagonist that may be clinically useful for the treatment of cardiovascular diseases such as hypertension. PMID- 8411774 TI - Pharmacological studies on cutaneous inflammation induced by ultraviolet irradiation (1): quantification of erythema by reflectance colorimetry and correlation with cutaneous blood flow. AB - This study was conducted to quantify the intensity of ultraviolet (UV) erythema in guinea pigs, a method for evaluating anti-inflammatory drugs, and to clarify any correlation of erythema with cutaneous blood flow. Skin color and cutaneous blood flow in non-administered and indomethacin-administered animals were measured by a colorimeter and a laser Doppler flowmeter over time after UV irradiation treatment. Skin color was indicated by a XYZ colorimetric system and L*a*b* color space. In either colorimetric system, the values of two indices, x and y or a* and b*, increased along with the intensification of erythema. The increase in the chroma (C*) value calculated from a* and b* was UV-dose dependent. This value was significantly suppressed by indomethacin 0.5-4 hr after irradiation, and it was found to be a clear and sensitive index for evaluating the suppressive effect of drugs. Cutaneous blood flow also increased with UV irradiation. Indomethacin significantly suppressed this increase 2-3 hr after UV irradiation. The changes of cutaneous blood flow correlated with those of C*. These results suggested C* was a suitable parameter to quantify UV erythema, and the change of skin color in UV erythema reflected the change of cutaneous blood flow. PMID- 8411775 TI - Possible involvement of ATP-sensitive K+ channels in the inhibition of rat central adrenergic neurotransmission under hypoxia. AB - By using rat brain cortical slices preloaded with [3H]norepinephrine, we examined whether ATP-sensitive K+ channels are involved in altered adrenergic neurotransmission during hypoxia. The tritium overflow evoked by transmural nerve stimulation (TNS) was significantly inhibited at 5 min of hypoxia and reached the maximum inhibition at 20 min. The inhibition of the TNS-evoked tritium overflow under a 20-min hypoxia was reversed by subsequent reoxygenation and was concentration-dependently antagonized by glibenclamide (0.1 and 1 microM). 86Rb+ efflux was increased after introduction of hypoxia and reached the peak value at about 20 min, which was concentration-dependently antagonized by glibenclamide (0.1-10 microM). Hypoxia decreased cortical ATP content. Linear correlations were mutually observed among the changes by hypoxia in the TNS-evoked tritium overflow, tissue ATP content and 86Rb+ efflux. The spontaneous tritium outflow was inhibited only after hypoxic periods of more than 16 min, the inhibition being reversed by reoxygenation and antagonized by 1 microM glibenclamide. These results suggest that the inhibition of rat central adrenergic neurotransmission during hypoxia may be associated with an activation of ATP-sensitive K+ channels. PMID- 8411776 TI - In vitro effects of the new calcium antagonist lacidipine. AB - The effects of lacidipine (LC), a new dihydropyridine calcium antagonist, were studied in comparison with those of nifedipine (NF) in isolated arteries of the dog (DG-AR) and isolated aorta (GP-AO), left and right atria (GP-LA, GP-RA) and ventricular papillary muscles (GP-PM) of the guinea pig. In DG-AR precontracted with high K+, LC and NF produced a concentration-dependent relaxation. The relaxant effect of LC was most potent in the basilar artery. The calcium antagonistic effects of LC was 8.7 and 2.1 times more potent than those of NF, in GP-AO and GP-LA, respectively. Thus, LC was about 4 times more selective towards vascular smooth muscles than NF. The negative chronotropic effects in GP-RA and the negative inotropic effect in GP-PM of NF were more pronounced than those of LC. NF was more potent in inhibiting the action potential of GP-PM than LC both in normal polarized and depolarized conditions. The effects of LC were long lasting. These results suggest that LC is a potent, highly vascular-selective calcium antagonist with little cardiodepressant effects and as such may be suitable for the treatment of hypertension. PMID- 8411777 TI - Modification by hydroxyl radicals of functional reactivity in rabbit lingual artery. AB - To understand the direct involvement of hydroxyl radical (.OH) in the modification of functional reactivity in isolated rabbit lingual artery ring preparations, this study was undertaken to examine the effect of .OH generated from dihydroxy fumarate (DHF) plus Fe(3+)-ADP or from H2O2 plus FeSO4. When vasodilators (acetylcholine and nitroglycerin) were given after the .OH generating system was removed from the organ chamber, the earlier .OH exposure produced an attenuation of the ring relaxation induced by acetylcholine but not that by nitroglycerin. Moreover, the earlier .OH exposure attenuated caffeine induced contraction and depressed the phasic response, but potently enhanced the tonic response of norepinephrine-induced contraction. Both the enhanced tonic response of KCl-induced contraction produced by earlier .OH exposure and norepinephrine-induced contraction was inhibited by nisoldipine. These results are consistent with the view that .OH radicals can potentiate the voltage dependent influx of Ca. It is also postulated that .OH may damage sarcoplasmic reticulum (SR) function in the smooth muscle cells, thus reducing Ca release from the SR (this may be reflected by the attenuation of the phasic response), and may selectively attenuate endothelium-dependent relaxation as opposed to endothelium independent relaxation. PMID- 8411778 TI - Effect of FK409, a novel nitric oxide donor, on acute experimental myocardial ischemia. AB - The anti-ischemic heart effect of (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]- 5-nitro 3-hexenamide (FK409), a novel nitric oxide donor, was studied in dog and rat preparations in vivo and in vitro. In anesthetized dogs with partially occluded coronary artery that were subjected to atrial pacing at a constant blood pressure, FK409 (1-100 micrograms/kg, i.v.) suppressed the ST-segment elevation on epicardial electrocardiograms. Glyceryl trinitrate (GTN; 10, 32 micrograms/kg) or dipyridamole (1000 micrograms/kg) failed to suppress the ST-segment elevation, although continuous i.v. infusion of GTN (32, 100 micrograms/kg/min) was effective. FK409 also suppressed the ST-segment elevation induced by methacholine in anesthetized rats by both i.v. (10, 100 micrograms/kg) and intraduodenal (i.d., 100, 1000 micrograms/kg) injections, while GTN (100 micrograms/kg, i.v.; 1000 micrograms/kg, i.d.) was effective only by the i.v. route. FK409 (0.32 microgram/kg/min, i.v.) and GTN (10 micrograms/kg/min) increased the blood flows of the endomyocardium (ENDO) and the epicardium (EPI) and the flow ratio of ENDO/EPI in the ischemic zone in anesthetized dogs with occluded coronary artery. Furthermore, in isolated dog vascular preparations, FK409 (4.6 x 10(-10)-4.6 x 10(-7) M) had a greater vasorelaxing effect on the large coronary artery [2.0-2.5 mm outer diameter (od)] than on the small coronary artery (0.3-0.5-mm od) or the saphenous artery. The results suggest that FK409 protects against acute experimental myocardial ischemia through relaxation of the large conductive coronary artery, and may be a useful oral drug for the treatment of angina pectoris. PMID- 8411779 TI - Involvement of brain serotonergic function in lidocaine-induced convulsions in mice. AB - Influences of drug-induced manipulations of central serotonergic function on lidocaine- and pentylenetetrazol (PTZ)-induced convulsions were examined in mice. Agents that suppressed serotonergic transmission increased, whereas drugs that facilitated serotonin (5-HT) function decreased the incidence of lidocaine induced convulsions. These treatments had similar influences on the incidence of PTZ-induced convulsions. Lidocaine (10(-5)-10(-3) M) reduced the stimulation evoked [3H]5-HT release from cortical slices, followed with an increased spontaneous [3H] overflow at higher concentrations. These results may suggest that brain 5-HT neurons are causally involved as inhibitory neurons in lidocaine induced convulsions as in the case of PTZ-induced convulsions. PMID- 8411780 TI - Typus melancholicus measured by a questionnaire in unipolar depressive patients: age- and sex-distribution, and relationship to clinical characteristics of depression. AB - We investigated the age- and sex-distribution of Typus melancholicus (TM) score by Kasahara's scale and the relationship of that score to various clinical characteristics in 110 unipolar depressive outpatients. The following results were obtained: The TM score in depressives is always constant regardless of age or sex, and has no relationship to clinical characteristics such as age, onset age, severity of depression, previous episodes, stress of life events, existence of melancholia, and the 4-month outcome. Patients with a higher TM score had no history of previous hypomanic episodes, while patients with a lower TM score had a significantly lower prevalence of early parental loss compared to those with moderate and higher TM scores. No difference in the prevalence of depressive family history was found between patients with different TM scores. PMID- 8411781 TI - CNS changes in DRPLA with dementia and personality changes: CT, MR and SPECT findings. AB - CNS changes in a case of DRPLA associated with dementia and personality changes were observed by CT, MR and SPECT. Brain CT and MR of the patient revealed progressive cortical atrophy which was recognized in parallel with the clinical course of the progression of dementia and personality changes. SPECT using 123I iodoamphetamine (IMP) disclosed a diffuse low perfusion of the cerebral cortex, especially in the frontal and temporal lobes. These findings suggest that the dementia and personality changes in this case might be concerned with a dysfunction of the cerebral cortex. PMID- 8411782 TI - Hot water epilepsy with pineal cyst and cavum septi pellucidi. AB - A case of reflex epilepsy accompanied by pineal cyst and cavum septi pellucidi induced by hot water bathing is presented. The patient is a 25-year-old male who has had six episodes of convulsions during the last three years. The seizures were diagnosed as a complex partial epilepsy followed by generalized tonic seizures. We have succeeded in recording his electroencephalogram (EEG) during convulsions. Moreover, a magnetic resonance imaging (MRI) study revealed a pineal cyst and cavum septi pellucidi. This is quite a rare form of adult hot water epilepsy accompanied by intracranial malformations. PMID- 8411783 TI - The effect of phenobarbital on the multiple neuronal activity in the hippocampus of El mice. AB - Phenobarbital (PB) (5, 25 and 50 mg/kg), which was administered intraperitoneally to evaluate the effect of it on El mice, inhibited convulsive seizures dose dependently. On the other hand, microelectrodes were implanted continually into the hippocampus of El mice, after which changes in the neuronal activity caused by a seizure were studied, as well as the effect of PB (5, 25 and 50 mg/kg) on the neuronal activity. The neuronal activity increased during the convulsive seizure when compared to the neuronal activity as the mice moved about freely. Further, after the convulsive seizure, five or six groups of bursts were seen periodically. PB reduced the neuronal activity dose-dependently judging by the number of bursts in the initial group of bursts. PMID- 8411784 TI - Sexual intercourse as a precipitating factor of transient global amnesia. AB - A Japanese female case of transient global amnesia (TGA), who developed an amnesic episode after sexual intercourse, is reported. The mechanism of how TGA in the present case is precipitated by sexual intercourse could be explained by the elevated blood pressure during sexual intercourse that drops rapidly, causing a temporal ischemia in the distribution of the basilar artery. Furthermore, the importance of sexual intercourse as a precipitating factor of TGA is stressed. PMID- 8411785 TI - Time course of dopamine-D2 and serotonin-5-HT2 receptor occupancy rates by haloperidol and clozapine in vivo. AB - In vivo occupancy of dopamine-D1, D2 and serotonin-5-HT2 receptors by haloperidol 10 mg/kg and clozapine 20 mg/kg were studied. Rats were injected intravenously with [3H]-YM-09151-2, [3H]-SCH23390, or [3H]-ketanserin 10 min after the administration of the tested drugs. Fifteen to 240 min after the ligand injection, the receptor occupancy rates of the drugs in the striatum and frontal cortex were calculated. Clozapine demonstrated the higher 5-HT2 and lower D2 occupancies in the respective regions. A dose-response analysis of D2 and 5-HT2 receptor occupancy by the drugs consolidated the higher 5-HT2 binding affinity of clozapine in comparison with haloperidol. The present methodology may serve as an accurate tool to evaluate the peculiarity of various antipsychotics. PMID- 8411786 TI - Five-year follow-up study of nursing home residents using the behavioral rating scale. AB - We followed up 109 elderly nursing home residents for 5 years using the modified Stockton Geriatric Rating Scale (SGRS). They were divided into 3 groups according to their walking ability, prolonged bedrest, walking with aids and walking by themselves. The scores of physical disability and apathy and the total score were in direct proportion to the degree of walking ability, and the score of communication failure of prolonged bedrest was significantly higher than those of other 2 states in the initial examination. The percentage of mortality of prolonged bedrest was significantly higher than those of other 2 groups. Physical disabilities of all the groups had deteriorated during the follow-up period, but only communication failure of walking with aids and the total score of walking by themselves had aggravated. It is considered that using of the behavioral rating scale such as the modified SGRS is conformative in caring of the elderly. PMID- 8411787 TI - Factor structure of Zung self-rating depression scale for workers. AB - The Zung self-rating depression scale (SDS) was applied to workers of a railway company. A total of 1,784 night-shift workers responded to the questionnaire and 1,394 daytime workers were set as the control group. The average age of the subjects was 39.9. The SDS index decreased with age. A factor analysis was conducted and two factors were obtained having an initial eigen value over 2.0. Factor 1 was the group of 8 positive items, accounting for 18.2%. Factor 2 was the group of 8 depressive items, accounting for 18.1%. Four items including "loss of weight," "constipation," "good appetite," and "partner coping" did not construct a distinct factor. The factor structure of SDS items of the night-shift workers was similar to those of the control group. PMID- 8411788 TI - A validation study of the Parental Bonding Instrument in a Japanese population. AB - Parker, Tupling & Brown's Parental Bonding Instrument (PBI), a self-rating scale for the measurement of perceived rearing attitudes of parents, was translated into Japanese and distributed to final-year high school students and to their parents. For each PBI score, ratings of each parent, made independently by family members, were weakly but significantly correlated. The social desirability score showed only a modest correlation to PBI scores. A factor analysis of the data, limiting the number of the factors retained to two, resulted in factor loading patterns similar to those reported by Parker, Tupling & Brown. PMID- 8411789 TI - Schizophrenia with pericentric inversion of chromosome 9: a case report. AB - A case of schizophrenia according to the DSM-III-R criteria with pericentric inversion of chromosome 9 was reported. This case and the review of the literature indicate that the pericentric region of chromosome 9 might be one of the potential regions of interest for linkage analysis of psychiatric disorders. PMID- 8411790 TI - Acutely administered haloperidol has little effect on steady-state visual evoked potentials from pattern-reversal stimulations in treated schizophrenics. AB - The amplitude changes in steady-state visual evoked potentials (VEPs) from pattern-reversal stimulations at 30 min after an intramuscular injection of haloperidol were examined in 10 treated schizophrenics. The VEP amplitudes to hemi-field or full-field pattern-reversal stimulations with a standard check size were almost unchanged after the haloperidol injection as compared with those before the injection. However, while the VEP amplitudes to the full-field stimulations were significantly higher in the midline occipital portion than in the right occipital portion before the injection, no significant difference was observed after the injection. Further, the VEPs similarly responded to the change in the check sizes used in the full-field stimulations before and after the injection, both showing significantly lower amplitudes only at the large check size of 4.1 degrees in a visual angle than that of 1.0 degrees. These results indicate that acutely administered haloperidol has little effect on steady-state VEPs from pattern-reversal stimulations in treated schizophrenics. PMID- 8411791 TI - Differential effects of tight and loose 2-hour restraint stress on extracellular concentrations of dopamine in nucleus accumbens and anteromedial frontal cortex. AB - Changes in the extracellular dopamine (DA) concentrations were examined in the nucleus accumbens (NAS) and anteromedial frontal cortex (AMFCx) during either 2 hr loose or tight restraint stress by means of a microdialysis method. Loose restraint induced significant 100% and 30% increases in DA release in the AMFCx and NAS, respectively, and the increased DA levels returned to the control values despite continued stress. Tight restraint induced an almost constant 100-130% increase during stressing in the AMFCx and a 30% reduction in the NAS. Transient increases in DA release were observed in both regions soon after the cessation of loose, but not tight, restraint. PMID- 8411792 TI - Endogenous event-related potentials in obsessive character. AB - Fifteen healthy subjects with obsessive character (OC) and 15 control subjects were tested for endogenous event-related potentials using the auditory odd ball paradigm. A difference was found in the peak amplitude of the P200 component in response to both stimuli; the subjects with OC had smaller amplitudes than the controls, and the mean amplitude for the 120 to 200 ms latency range was smaller (negative shift) for the OC subjects. In the OC subjects, NA appears to be markedly induced to both rare and frequent stimuli; moreover, the OC subjects may show excessive reactions to selective attention as well as to the process of pattern recognition. PMID- 8411793 TI - Smooth pursuit eye movement dysfunction as a biological marker for prediction of disease courses of schizophrenia: a preliminary report. AB - We investigated the relationship between a smooth pursuit eye movement (SPEM) dysfunction and long-term disease courses of schizophrenia. Many schizophrenic patients without the SPEM dysfunction tended to show an acute onset of illness, undulating courses and relatively good outcomes. On the other hand, patients with cogwheel-like SPEM dysfunction tended to show a chronic onset, simple courses, relatively severe outcomes and negative symptoms. PMID- 8411794 TI - EEG changes with aging in adults with Down syndrome. AB - The EEGs of 40 subjects with Down syndrome (DS) aged 15 to 54 were investigated and compared with those of 42 mentally retarded subjects without DS (MR) and 47 healthy volunteers. The frequencies of the occipital alpha rhythms decreased with age in the DS subjects, and were significantly slower than the MR subjects over 35 years old. The slowing of alpha rhythm in the older DS subjects accompanied an increase of slow activity of 6-8 Hz, but did not relate to the deterioration of activities of daily life (ADL) as estimated by the GBS score. Twenty-one subjects with DS were examined by a CT scan as well, and showed no age-related morphometric changes. PMID- 8411795 TI - Agreement of visual scoring of sleep stages among many laboratories in Japan: effect of a supplementary definition of slow wave on scoring of slow wave sleep. AB - The purpose of this study was to elucidate the agreement of visual scoring of all night polysomnographic recordings among many scores from different laboratories. Ten scorers including the author from different laboratories in Japan scored the same paper recordings of two young male subjects. We calculated the agreement rate for each stage using an epoch by epoch analysis. In both records, the agreement rates for stages 2 and R were high; on the contrary, those for stages 3 and 4 were low. After adding a supplementary definition of high voltage slow wave in deep sleep, we scored the first NREM period of another subject. The mean agreement rate for stage 3 among 10 scorers was significantly higher than those of the two former subjects. However, the agreement for stage 4 did not change so much. This result demonstrates that there is much interrater (laboratory) variability of visual scoring, especially in slow wave sleep. When the result of automatic scoring is compared to that of the visual scoring to evaluate the reliability of automatic scoring, these findings must be considered. PMID- 8411796 TI - [Immunosuppressive therapy in renal transplantation]. AB - In 1960, the beneficial effect on allograft survival of 6-mercaptopurine (6-MP) was demonstrated. In 1961, azathioprine, a less toxic derivative of 6-MP, was synthesized and soon became the main form of maintenance therapy. Around the same time, clinical studies of the effects of corticosteroids on the immune system were being conducted and successful reversal of rejection with cortisone was recognized. Subsequently, steroids were added to azathioprine and this regimen constituted maintenance immunosuppressive therapy in most centers for over 20 years. However, these first generation immunosuppressive medications are the cause of the majority of complications following renal transplantation. Both azathioprine and steroids effect nearly all immunologic and inflammatory responses and may predispose to infection and certain malignant diseases. Despite refinement in the use of these agents with improved patient survival, little improvement in graft survival was appreciated. The second generation immunosuppressive agents, antilymphocyte globulin (ALG), emerged in the form of polyclonal antibodies derived from xenotypic antisera and directed against the entire cellular response. Unfortunately, many limitations have been encountered with respect to the administration of ALG. Cyclosporine A is more specific and attacks a subset of the lymphocyte population, the T helper lymphocytes. When cyclosporine A is used in combination with prednisone, the immunologic cycle that causes rejection is interrupted twice by virtue of the fact that prednisone prevents the production of IL-1 by macrophages and cyclosporine A with the production of lymphokines, especially IL-2 (T-cell growth factor). Although cyclosporine A is at present better immunosuppressive drug to azathioprine, it is not without a number of side-effects. Nephrotoxicity, however, is the most worrying complication in patients receiving cyclosporine A. Of most importance to the urologist, however, is the interaction with cyclosporine A by many of the drugs used to treat various urologic infections. The advent of hybridoma technology which has become so important to the urologic oncologist has also impacted upon transplantation and resulted in the development of a new generation of antilymphocyte antibodies. The bulk of clinical experience has been obtained with OKT3, a monoclonal antibody directed against the T3 antigen complex found on all T-lymphocytes. A large multi-center experience has showed the remarkable efficacy of OKT3 in providing rapid depletion of peripheral blood T cell levels and reversal of established rejection not responded to conventional high-dose steroid therapy and in reducing the frequency and delay of the onset of rejection reactions. PMID- 8411797 TI - [Studies on the existence of renin in the human testis]. AB - Recent reports have shown the existence of renin in Leydig cells of the rat testis and immunoreactive renin activity in the human testis. But we could not rule out the possibility that the renin found in the testis is contamination of plasma components by endocytosis. In the present study we tried to demonstrate the renin production in the human testis by the following experiments. At first, we measured the plasma renin activity and testosterone in the internal spermatic vein simultaneously after treatment with hCG and obtained the results that suggested hCG-induced renin secretion in the human testis. Next, we showed the hCG-dependent renin production from the cultured human Leydig cell tumors in vitro. Thirdly, we demonstrated the existence of renin mRNA in the human testisusing reverse transcriptase-polymerase chain reaction (RT-PCR). Fourthly, we detected significant signals of human renin mRNA in the human testicular tissues by in situ hybridization. From the above results, it was demonstrated that the human renin gene was hCG-dependently expressed in the human testis. PMID- 8411798 TI - [Characteristics of bladder tumors showing positive urinary cytology--respective analysis in differentiated and undifferentiated type]. AB - To study the characters of bladder tumors which showed positive urinary cytologies, the results of urinary cytology of untreated bladder tumor patients in our hospital during 10 years between July 1980 and June 1990 were evaluated according to the degree of tumor differentiation, namely differentiated type which did not contain grade 3, and undifferentiated type which contained grade 3. Concerning the differentiated tumors, positive rate of cytology was 37.3% on the whole. Positive rates of multiple tumors or tumors around internal urethral orifice were significantly high, and those of tumors larger than 3 cm, tumors located on posterior wall, left wall or throughout the entire surface, were higher than the overall positive rates. Repeated examinations of yielded increased rates of positive cytology results. Concerning the undifferentiated tumors, the positive rate of cytology was 90.0% on the whole, was not influenced by tumor sizes or locations, and was consistently observed irrespective of the number of times of cytologic examination. PMID- 8411799 TI - [Transition of diagnosis and treatment for impotence]. AB - Statistic study was made on 685 outpatients with erectile dysfunction during 13 years periods from 1979 to 1991 in the Department of Urology, Takamatsu Red Cross Hospital. The age distribution showed the highest frequency in the thirties decade (27.9%). 1. Diagnosis Nocturnal penile tumescence monitoring was the only method to distinguish organic impotence from functional impotence during the first 5 years. From 1984 we have been able to diagnose corporal veno-occlusive insufficiency (CVI) by papaverine test and dynamic infusion cavernosometry and cavernosography. Measurement of penile brachial index and pelvic angiography were performed to diagnose arterial insufficiency (AI). In neurological examination we have measured bulbo-cavernosus reflex latency and nerve conduction velocity of the dorsal penile nerve and performed microvibration measurement and sweat spots test. All diagnostic methods were established in 1987 and we have been able to classify about 80% patients. We classified 305 patients during the last 5 years from 1987 as follows: psychogenic; 31.1%, CVI; 20.0%, AI; 7.9%, neurogenic; 7.9%, others; 14.1%, drop-out and unknown; 19.0%. 2. Treatment In the beginning only counseling and drug therapy were performed. However we lately performed various suitable methods for individuals based on their diagnoses. We performed counseling, drug therapy and intracavernous injection of vasoactive drug therapy for recovery of spontaneous erection (vascular training) for psychogenic IMP. The efficacy rate of counseling was low (30.4%) but that of drug therapy was 41.6%, and that of vascular training, 64.2%. For CVI without other factors we performed venous surgeries. Only about 25% of them perform sexual intercourse with or without self injection of vasoactive drug (self injection).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411800 TI - [An experience with a continent appendix stoma]. AB - The submucosally embedded in situ appendix guarantees an ideal continence mechanism in patients with ileocecal urinary reservoirs. To date this modification of the Mainz pouch technique was performed successfully in 10 patients between September 1990 and October 1992. Pouch capacity ranged from 350 ml to 900 ml (mean: 565 ml), frequency of intermittent self-catheterization ranged from 3 to 7 times per day (mean: 4.4-5.4 times) and complete urinary continence without difficulty of self-catheterization was obtained in eight patients. Two patients with stomal stenosis needed to indwell a catheter. The follow up periods ranged from 3 to 24 months (mean: 14.7 months). The main advantages of appendix stoma technique are shorter operation time, a short ileal segment, no risk of nipple gliding or prolapse, no need for staples, thereby decreasing significantly the risk of stone formation, and facilitating easy catheterization. This method seemed to be valuable for continent urinary reservoir assuring patients of high quality of life. PMID- 8411801 TI - [Use of ultrasonography in the bladder neck suspension]. AB - From 1988 to 1992, 78 patients with genuine stress urinary incontinence underwent bladder neck suspension under ultrasonic monitoring. Tightness of suspension was adjusted by setting posterior urethrovesical angle to approximately 90 degree by transrectal ultrasonography during operation. Urinary continence was achieved in 68 of 78 patients. In the remaining 10 patients, slight incontinence recurred within 6 months after operation. In 60 patients undergoing postoperative chain cystourethrogram, the posterior urethrovesical angles set during operation were about the same as those after operation. In 39 patients undergoing uroflowmetry under the condition that micturition volume was 200 ml or more, urinary flow rate did not decrease after operation. In 20 patients whose posterior urethrovesical angles were measured by transperineal and transabdominal as well as transrectal ultrasonography, angles measured by each ultrasonography were almost identical and neither manipulation could change the configuration of the bladder neck. The posterior urethrovesical angles set during operation were kept postoperatively and provided proper tightness of the suspension suture to achieve urinary continence without any difficulty of urination. Therefore, a posterior urethrovesical angle can be set by transperineal and transabdominal ultrasonography as well as transrectal ultrasonography. While transrectal approach provides the clearest image among the three approaches, transperineal and transabdominal approach carried out with a transabdominal convex or sectorial probe are more convenient than transrectal approach which needed a special probe. PMID- 8411802 TI - [Urinary incontinence following total prostatectomy--evaluation by urodynamics and urethrography]. AB - Urinary incontinence following total prostatectomy was evaluated in 10 patients by urodynamics and lateral urethrography. The pathological stage of the tumors in these patients was pT2 in 5 patients, pT3 in 3 patients and pT4 in 2 patients. Urinary incontinence was present in 8 patients, of whom 7 were stress incontinence and one was urge and stress incontinence. The severity of incontinence of these patients was mild in 5 patients, moderate in 3 and severe in 2. The incontinence was severer in the patients with the tumors of pT3 or pT4 than in the patients with the tumors of pT2. But the severity of incontinence was not related to the pathological grade or resected weight of the tumors. The bladder capacity and bladder compliance were 214.9 ml and 14.3 ml/cmH2O in average, respectively, and was not related to severity of incontinence. The statistical significant differences between continence or mild incontinence patients and moderate or severe incontinence patients were found for the mean functional profile length (2.04 versus 1.37 cm, respectively; p < 0.05) and maximum urethral closure pressure (42.3 versus 17.3 cmH2O, respectively; p < 0.05). But some patients with continence or mild incontinence demonstrated low values in either parameters. No statistical difference was found between continence and mild incontinence patients. On lateral urethrography, the posterior urethrovesical angle was not correlated with the severity of incontinence. PMID- 8411803 TI - [Study on the cellular proliferation and endocrine activity of the parathyroid gland in hyperparathyroidism]. AB - Flow cytometric DNA analysis was performed in 21 cases of primary hyperparathyroidism (19 adenomas in 19 cases and 5 glands with primary hyperplasia) and 24 glands with secondary hyperplasia. Tissue intact-PTH concentration was measured in another 41 glands with secondary hyperplasia. Glands with adenoma had a higher proliferative index than normal glands or glands with secondary hyperplasia. In the adenoma group, there was a negative correlation between proliferative index and age, an indicator of the tumorous nature of adenomas. Daily urinary calcium excretion in case of adenoma and tissue intact-PHT concentration in glands with secondary hyperplasia also showed correlations with proliferative index, which demonstrates a close relationship between cellular proliferation and endocrine activity. Gland weight and proliferative index had no significant correlation in both groups. Therefore, another mechanism besides cellular proliferation and endocrine activity, such as number of non-functional parathyroid cells, should be considered. PMID- 8411804 TI - [A study of the quantitative dual-parameter flow cytometry analyses of cellular antigens on transitional cells for the diagnosis of bladder cancer]. AB - To establish a quantitative dual-parameter flow cytometry (FCM) analysis of cell surface antigens, possible obstacles caused by contaminated leucocytes in a specimen and staining and measuring conditions were investigated using human bladder cancer cell lines, 5637, T24 and SW1710. The first monoclonal antibody (MoAb) used to select urothelial cells in a specimen was applied with the second MoAb used to discriminate between normal and transformed urothelial cells. MoAbs Due AUT 2 and CD45 appeared to be suitable for the selection of urothelial cells, while Due ABC 3 and Due ABC 5 were applied to detect transformed cells. Tumor cell-leucocyte suspension was simultaneously stained with combinations of these MoAbs. The results demonstrated that Due AUT 2 and CD45 effectively eliminated contaminated leucocytes by means of positive and negative selection of the urothelial cells, respectively. Based on these experiments, dual-parameter FCM analyses of bladder washing from 5 patients with bladder cancer were performed using MoAbs Due AUT 2 and Due ABC 3. The results indicated that by dual-parameter FCM distinct antigenic features of transitional cells could be investigated even if considerable amounts of contaminated leucocytes were present. The clinical impact of this approach is a subject of ongoing trials. PMID- 8411805 TI - [Studies of argyrophilic nuclear organizer region proteins in renal cell carcinoma. Its significance as a marker of proliferative activity]. AB - Tumor grading protocols in renal cell carcinoma (RCC) have met with only limited success of prognostic value. The increase in argyrophilic nucleolar protein (AgNOR) count of a cell has been reported to reflect rDNA transcriptional activity and state of cellular activity. Therefore, the relationship between number of the AgNOR in nucleus and proliferative activity or clinical course in patients with RCC was evaluated. Using 6 cell lines derived from RCC patients and established in our institute, the number of AgNOR and doubling time of the cell were compared. These two values had statistically high negative correlation; a coefficient of correlation was -0.88. The AgNOR values in the cells of RCC (2.22 +/- 0.81) in 45 patients were significantly higher than those of normal proximal tubules (1.54 +/- 0.21) (p < 0.05). And, the numbers were increased with the progress of grades in RCC; 1.70 +/- 0.33 in Grade 1, 2.12 +/- 0.74 in Grade 2 and 3.19 +/- 0.19 in Grade 3. Moreover, the numbers of AgNOR were higher in infiltrating type such as sarcomatoid type or INF gamma. However, no significant relation in the AgNORs numbers was observed in subcell type of RCC. The patients with RCC having a counts of 2.5 or more of AgNOR showed a significantly poor prognosis. These results suggest that the determinations of the AgNOR numbers have a value to evaluation of the tumor proliferative activity in RCC patients. PMID- 8411806 TI - [Preoperative collection of autologous blood with recombinant human erythropoietin therapy]. AB - The major problems in liquid storage of donated blood are the limitation of blood volume preserved and the time consumed. We studied on the usefulness of utilization of recombinant human erythropoietin (rH-EPO) for preoperative collection of autologous blood. Patients were divided into 2 groups: Group 1 (12 patients) received 3000 unit of rH-EPO intravenously every day. Group 2 (10 patients) was not treated with rH-EPO. All the patients were given ferrous sulfate 100 mg of iron orally every day. As a rule, 400 ml blood was donated two or three times a week for rH-EPO treated group, while, for control group 400 ml blood was collected once a week. Some patients did not donate 400 ml because of their critical condition. The blood volume preserved became 780.9 +/- 284.1 ml in Group 1, and 910.8 +/- 263.9 ml in Group 2. However, the number of days until operation were significantly shorter in Group 1 (8.7 +/- 3.6 days) than in Group 2 (26.2 +/- 11.2 days). Hemoglobin and Hematocrit levels recovered promptly in rH EPO treated group. In control group, 2 patients complained of nausea and vomiting caused by severe anemia. On the other hand, no one with rH-EPO therapy did not show any side effects and complications. Our study indicates that the administration of rH-EPO enables us to collect an adequate volume of autologous blood preoperatively in a shorter period of time. PMID- 8411807 TI - [Reassessment of the sonographic criterion of prostate cancer--nonspecificity of a hypoechoic lesion]. AB - Of 71 patients who underwent ultrasound guided transrectal needle biopsy at the urology clinic in Kitasato University Hospital 19 (26.8%) patients were found to have prostate cancer. The positive predictive value for transrectal ultrasonography (hypoechoic lesion) was disappointing-only 30% (18/60). While hypoechogenicity has been increasingly accepted as the most sensitive ultrasonographic indicator of malignancy, the specificity of hypoechogenicity is related to benign structures that may also appear hypoechoic. Sonographic detection of prostate cancer is also confounded by the presence of iso- and hyperechoic tumors. Experienced sonographer should be alert to this limitation of the current technology of transrectal ultrasound. The advent of the Biopty gun enabled us to take multiple biopsy cores easily and quickly. Liberal use of random systematic biopsy in addition to directed cores from a hypoechoic lesion should enhance the detection of prostate cancer regardless of the tumor echogenicity. PMID- 8411808 TI - [Experimental reconstruction of the urinary bladder using atelocollagen sponge]. AB - An experimental reconstruction of the urinary bladder using specially designed bovine cross linked atelocollagen sponge was performed after partial cystectomy in 12 male rabbits. Bladder reconstruction with hybrid type biomaterials using both atelocollage sponge and cultured autologous cells were performed in 5 rabbits (the 1st group). The autologous cells were collected from the mucosa and muscular layer of the urinary bladder and were seeded on the atelocollagen sponge before reconstruction. Reconstruction using atelocollagen sponge without autologous cells were performed in the other 7 rabbits (the 2nd group). In 9 of these 12 rabbits, atelocollagen sponge was successfully implanted in the native urinary bladder. Histopathological findings revealed that seeded autologous cells and growth of surrounding host cells could be seen similarly in the atelocollagen sponge but these could not be differentiated by these could not be differentiated by routine histopathological techniques. In conclusion, these results showed a possibility of construction of artificial urinary tract using both atelocollagen sponge and autologous cells. PMID- 8411809 TI - [Demonstration of interleukin-1 (IL-1) as an autocrine growth inhibitor in renal cancer cell line, TC-1]. AB - In this report, we demonstrated that Interleukin-6 (IL-6) production could be induced by stimulating renal cell carcinoma cell lines, namely ACHN, Caki-1 and TC-1 cells with Interleukin-1 beta (IL-1 beta). IL-1 beta had no effects on cell proliferation in ACHN cells. However, IL-1 beta could suppress cell proliferation in Caki-1 and TC-1 cells. Flow cytometric cell cycle analysis by double staining method with propidium iodide and Proliferating cell nuclear antigen (PCNA) disclosed IL-1 beta caused cell accumulation at G1 phase. Fine granules were visualized in perinuclear area of TC-1 cells treated with IL-1 beta under microscopy. High electron density granules and spherically dilated rough endoplasmic reticula were observed by electron microscopic examinations. In TC-1 cell culture, IL-1 beta excretion into the supernatant was demonstrated by bioassay and ELISA. These results suggest that IL-1 beta functions as an "autocrine growth inhibitor" against TC-1 cells. Half-maximal inhibition of IL-1 beta and IFN-alpha was 6.5 pg/ml, and 720 U/ml, respectively for TC-1 proliferation and combination of these cytokines showed enhanced activity in cell growth inhibition. PMID- 8411810 TI - [Study on semen collection from patients with anejaculation by electrostimulation]. AB - Several methods to provoke ejaculations artificially have been used to treat patients with anejaculation. The purpose of this study is to investigate the clinical usefulness of electroejaculation for patients with anejaculation induced by spinal cord injury or surgical operation. Forty patients with spinal cord injury and one with anejaculatin after radical operation for rectal cancer underwent electroejaculation by Brindley's technique and/or modified Seager's technique. To reduce the electric energy consumed in patients, we applied bipolar pulses for modified Seager's technique instead of sine-wave current. Seminal emissions were obtained from 11 out of 18 patients by Brindley's technique and from 24 out of 29 patients by Seager's technique. From the patient with anejaculation after radical operation for rectal cancer, semen was obtained by modified Seager's technique. Most common complications were autonomic dysreflexia and pain. Since these complications disappeared soon after the termination of electrostimulation, this method was thought to be safer than intrathecal injection of neostigmine. Regarding the semen quality, it seemed that total sperm counts got worse as the interval after injury was extended. Not only in the chronic stage but also in the first one month after spinal cord injury, sperm motility was low. It was suggested that dysfunction of the testis and epididymis appeared in the early stage of spinal cord injury. PMID- 8411811 TI - [Renin expression in rat testis is controlled by gonadotropin]. AB - The effect of human chorionic gonadotropin (HCG) on renin concentration and renin mRNA was studied using the testis and kidney of rats. Forty-five rats were divided into three groups. The first group (25 rats) was used for short time course study with single injection of 150 IU HCG and the second group (10 rats) for long term injection of 150 IU HCG daily for 3 weeks. In both groups, serum and testicular sample were taken after 24 hours both nephrectomy in order to exclude the influence of renal renin and angiotensin. The third group (10 rats) was used to find the effect of HCG on renal renin. Plasma and testicular renin concentration were measured by radioimmunoassay of angiotensin I. The specificity of renin concentration was determined using a specific renin antibody. Renin mRNA of the kidney and testis were measured by a sensitive RNAase protection assay method. Testicular renin mRNA increased and reached the maximum level at 8 hrs after 150 IU HCG single injection. And also testicular renin concentration significantly increased and it showed the maximum levels at day 3. However long term HCG administration in rats showed higher in levels in testicular renin concentration, however renin mRNA level was not increased. Non-nephrectomized rats showed low levels of plasma renin concentration and no changes in mRNA after HCG administration. The results suggest that renin angiotensin results suggest that renin angiotensin system independently exists in the testis and testicular renin mRNA is increased by HCG administration. PMID- 8411812 TI - [A case of retroperitoneal fibrosis forming a pelvic tumor--a type of multifocal fibrosclerosis]. AB - We experienced a case of retroperitoneal fibrosis with formation of a pelvic tumor. The patient was a 55-year-old female who showed right-side hydronephrosis on examination and was admitted to this department for consultation. Retrograde pyelography clearly showed that the hydronephrosis was due to lower ureteral obstruction resulting from a pelvic tumor. Ultrasound, CT and MRI showed a tumor adhering to the right posterior surface of the bladder, and it was thought that the tumor was probably malignant. Transdermal/transvesical needle biopsy and transvaginal needle biopsy showed a minimal invasion and allowed the diagnosis of retroperitoneal fibrosis to be made. Ureterolysis was not carried out, and the patient was treated with steroid monotherapy. She was begun on 30 mg of prednisolone, and administration was continued for approximately 3 months while gradually decreasing the dosage. On completion of administration, the intrapelvic tumor had completely disappeared, showing that treatment was markedly effective. The patient had a history of an orbital pseudotumor, and it was concluded that this was a rare case coming under the heading of multifocal fibrosclerosis. PMID- 8411813 TI - [Quantitative electromyographic analysis of the external urethral sphincter using automatic decomposition electromyography on normal subjects]. AB - Motor unit action potentials (MUPs) of the external urethral sphincter muscle during cystometry were analysed on six normal males. Electrodes were of concentric needle type having the exposed tip surface of 0.07 mm2, and the potential changes were analysed by automatic decomposition electromyography (ADEMG), for isolation and characterization of unit discharges. Number of analysed units was as follows: Two at rest, 4 at first desire to void, and 6 at maximum desire to void. Number of the recorded units and the firing rate were increased along with the degree of bladder filling. Average MUP figures as follows: Amplitude of 206 microV, 8.3 msec. duration, and 5.4 Hz firing rate at rest; 246 microV, 9.7 msec and 7.3 Hz at first desire to void; 277 microV. 9.7 msec and 7.2 Hz at maximum desire to void, respectively. It is concluded that external urethral sphincter maintains its urinary continence by excitation of the neuromuscular units, and by the increase in the firing rate of individual MUP during bladder filling. PMID- 8411814 TI - [Quantitative electromyographic analysis of the human external urethral sphincter: effect of alpha-adrenergic agents]. AB - External urethral sphincter electromyography (EMG) during bladder filling was quantitatively analysed using automatic decomposition electromyography (ADEMG). These include 7 normal volunteers, 7 patients with outlet obstruction, 9 with sphincter dyssynergia, and 7 with pelvic nerve injury (PNI). Recruited number of participated motor unit action potentials (MUPs) at maximum bladder filling was significantly larger than that at rest in the normal volunteers, patients with outlet obstruction, and sphincter dyssynergia. In patients with PNI, MUPs were not increased. The MUP amplitude did not differ significantly among groups during bladder filling. In PNI group, MUP duration at rest tended to be larger than those in other groups. Firing rates at first desire to void and at maximum desire to void were significantly increased than that at rest in every groups except the PNI group. Effect of alpha-1 adrenergic blocking agent, phentolamine (5 mg intravenously), was examined on 3 patients with benign prostatic hypertrophy (BPH), and in 4 patients with sphincter dyssynertia. Effect of an alpha-1 adrenergic stimulating agent, norepinephrine (5 microgram/min intravenously), was also evaluated on 5 patients with PNI during bladder filling. Immediately after administration of phentolamine, number of MUPs was decreased in BPH and sphincter dyssynertia groups. But in 2 paraplegic patients having reflex bladder with sphincter dyssynergia, number of MUPs did not decrease. Even with the continuous intravenous administration of norepinephrine for the elevation of the blood pressure, number of MUPs did not increase in any PNI patients. Effect of intravenous administration of phentolamine to induce urethral pressure elevation and the recording of the evoked EMG from external urethral sphincter by electrical stimulation of the penile dorsal nerve was tested on 4 patients without PNI before and after the administration. Phentolamine lowered urethral pressure at membranous urethra, but no significant change of evoked urethral pressure response and EMG activities was observed. It is concluded that the decrease of MUP after phentolamine administration is due to the relaxation of periurethral part of external urethral sphincter, and clinical denervation supersensitivity of the external urethral sphincter after norepinephrine administration was not demonstrated in the PNI groups. PMID- 8411815 TI - [Transurethral ureterolithotripsy with rigid ureteroscope--efficacy of balloon dilation of ureteral orifice and intramural ureter]. AB - In order to minimize the late complications of transurethral ureterolithotripsy with a rigid ureteroscope such as distal ureteral stricture or vesicoureteral reflux, we have performed a balloon dilation of the ureteral orifice and the intramural ureter prior to insertion of a rigid ureteroscope in 62 cases with ureteral calculi between October 1986 and December 1991. A 7F balloon dilator (Bard Co. USA) was cystoscopically inserted into the ureter over a 0.038 inches Radifocus guide wire (Terumo Co., Japan) and then, a balloon was inflated to 15F size with saline using a 10 ml disposal syringe. Thereafter, an 11.5F rigid ureteroscope (Wolf Co., Germany) was inserted instead of a balloon dilator. The stone was disintegrated and removed by ultrasonic lithotripter, electrohydraulic lithotripter and/or basket forceps. The stone was removed completely in 45 cases (72.6%) and incompletely in 10 (16.1%). In 7 cases (11.3%), the stone removal was failed. Long-term follow up with an excretory urography obtained at 6 months or later in 31 cases showed no distal ureteral stricture. Voiding cystography was performed in 33 cases at a mean 2.8 months postoperatively and no vesicoureteral reflux was observed. In control group, 309 cases were treated with transurethral ureterolithotripsy without dilation of the ureteral orifice and the intramural ureter between the same period. A complete removal was achieved in 262 cases (84.8%) and an incomplete removal in 16 (5.2%). 31 (10%) was failed. Distal ureteral stricture due to this procedure was found in 3 cases (2.1%) in 141 long term followed up cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411816 TI - [Experimental studies on urinary tract infection--bladder and kidney lodgement in mice by strains of Staphylococcus epidermidis]. AB - Eight strains of Staphylococcus epidermidis were isolated from clinical specimens and were subjected to the tests for encapsulation, slime-production, mouse virulence, capsular-typing and adherence to the plastic surface, the renal cells and the bladder epithelial cells. With the intraperitoneal injection of these strains into mice, three encapsulated strains were observed in mouse-virulent, while two slime-producing and three unencapsulated strains were mouse-avirulent. In ultra-thin sections of mouse-virulent strains, capsules were demonstrated by electron microscopy, however no capsule was seen around the walls of mouse avirulent strains. On the other hand, slime-producing strains were shown to have higher adherence rate to plastic surface, compared to encapsulated strains. The direct adherence rate to the bladder epithelial cells was greater in the encapsulated strains than the slime-producing strains, although there was no difference to the renal cells. Also, cell suspension containing 10(7) colony forming units of these strains were injected intravenously or intravesically into mice. Of these encapsulated strains showed kidney and bladder lodgement with in 28 and 21 days after intravesical injection, respectively, although slime producing and unencapsulated strains could only be obtained for 7 days in the kidney and 3 days in the bladder. However, there was no difference in the internal organs lodgement of the strains after intravenous injection into mice. These experimental results suggested that the encapsulation of the strains would be an important factor for bladder lodgement, however, significant effects were not observed with the slime-producing and unencapsulated strains. PMID- 8411817 TI - [Assessment of the quality of life of prostate cancer patients]. AB - Assessing the QOL in cancer clinical trials is becoming increasingly important. However, a suitable device of assessment of QOL has not been developed yet for prostate cancer patients in Japan. We tried to assess the QOL of prostate cancer patients using the EORTC questionnaire translated in Japanese, and examined its validity and reliability. Thirty-six patients filled in a questionnaire. The reliability of this device was confirmed by the results of test-retest reproducibility. Good correlation was shown between the results and patients performance status, and between the results and clinical stages, which support the validity of the device. As for the results of assessment of QOL, these patients were severely damaged in sexuality. Those factors of functional status, physical symptoms and fatigue/malaise were closely related to disease activity and clinical stage. PMID- 8411818 TI - [Laparoscopic nephrectomy; results of initial 14 cases and procedure for renal cell carcinoma]. AB - Since July 1991, we have performed laparoscopic nephrectomy for removal of damaged kidneys in thirteen patients. In addition, we recently developed a new procedure of laparoscopic nephrectomy for renal cell carcinoma and successfully removed the kidney with renal cell carcinoma. Patients' age ranged from 18 to 80 years with the average being 50 years. There were 8 male and 6 female patients. Original disease was calculous disease in 5 patients, vesicoureteral reflux in 4, renovascular disease in 2, ureteropelvic junction obstruction in 1, ectopic ureter in 1 and renal cell carcinoma in 1. The mean size of the diseased kidney was 9 x 5 x 4 cm in the initial 13 cases and 13 x 7 x 6 cm in the case with renal cell carcinoma. The size of the tumor was 2.5 x 2 x 2 cm. In the first 13 patients, nephrectomy was performed as described previously. In the patient with renal cell carcinoma, a new procedure was applied. In the new procedure, five trocars were introduced into the abdominal cavity through the lateral abdominal wall. The kidney was removed en bloc together with the adrenal gland, perirenal fat and Gerota's fascia in the same fashion as traditional radical nephrectomy. Thirteen kidneys were successfully removed, but one failed because of dense adhesion to the surrounding tissue. Three of the 13 patients required additional laparotomies to control bleeding or to remove missed stone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411819 TI - [Urokinase-type plasminogen activator antigen as a prognostic factor in bladder cancer]. AB - The relationship between the urokinase-type plasminogen activator (u-PA) content in extracts of cancer tissues and the histological grade or the extent of the primary tumor as prognostic factors in bladder cancer were examined in 55 cases of bladder cancer. The patients were divided into 2 groups, high u-PA (8 ng/ml of protein and more) and low u-PA (less than 8 ng/mg of protein) groups. The incidence of vascular invasion was 37% in the high u-PA group, and 7.1% in the low u-PA group (p < 0.05). The incidence of high u-PA group increased with the grade of cancer and the extent of the primary tumor. The 3-year survival rates were 87.3% in low u-PA group and 42.6% in high group (p < 0.005). The survival rates of the patients in the high u-PA group were lower than in the low group, by grade or stage. In a multivariate analysis, the prognostic value of u-PA antigen content was the same as that of tumor grade and stage. Therefore, the content of u-PA may be a useful prognostic marker for bladder cancer in addition to tumor grade and stage although the prognosis of the patients with bladder cancer is considered to be determined by many factors. PMID- 8411820 TI - [A study of testicular tumor on magnetic resonance imaging]. AB - Magnetic resonance imaging (MRI) was performed on 13 testes of 12 patients with testicular abnormalities. In all cases, scrotal palpation and initial clinical course had suggested malignant testicular tumors. The normal testes showed homogeneous intermediate signal intensity on T1-weighted images and high signal intensity on T2-weighted images such as the appearance of the corpus cavernosum penis. Ten patients proved to have testicular tumors. In eight of them, the tumors showed homogeneous intermediate signal intensity on T1-weighted images, while in the other two, these images showed heterogeneous signal intensity. In four patients, tumors were of homogeneous low signal intensity on T2-weighted images, while in the other six, these images showed heterogeneous signal intensity. The rate of correspondence about local staging of testicular tumor between MRI and pathological diagnosis was 70 percent. In three patients whose testes were undoubtedly malignant according to clinical findings and examination data, MRI enabled us to diagnose one case of testicular atrophy. However, MRI suggested benign disease in the other two patients; nevertheless, surgery was performed as indicated by the other findings. PMID- 8411821 TI - [DNA ploidy of testicular germ cell tumors in childhood; difference from adult testicular tumors]. AB - It is well known that there are various differences in the biological characteristics and clinical behavior between prepubertal testicular germ cell tumors and adult ones. We analyzed the nuclear DNA ploidy of testicular tumors in childhood using DNA flow cytometry for clarifying those biological features and shedding some insights in the pathogenesis of testicular germ cell tumors. Formalin-fixed, paraffin-embedded specimens of primary tumors taken from 9 boys with histological evidence of yolk sac tumors and 8 with prepubertal teratoma treated in our clinic were used for flow cytometry analysis with some modification of the Hedley's technique. The results were compared with those of adult testicular tumors which we previously reported. All specimens in children showed "DNA euploid"; DNA diploid in all teratomas and 6 yolk sac tumors, DNA tetraploid in other 3 yolk sac tumors. Neither distinct DNA aneuploidy nor DNA heterogeneity were detected in children. Our previous study proved that the vast majority of adult testicular tumors contain DNA aneuploid stemlines. Although prepubertal yolk sac tumor and teratoma are histologically identical with those in adults, this study apparently reveal the different DNA stemline ploidy in prepubertal testicular tumors compared with that in adult ones. It has been known that carcinoma in situ (CIS) of the testis is a precursor of adult testicular germ cell tumors and the CIS cells in precancerous state already shows aneuploid DNA histogram patterns. Moreover, CIS has never been observed in children. The current results are in agreement with the hypothesis that the pathogenesis of prepubertal testicular tumor is different from that of adult ones.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411822 TI - [Microsurgical vasovasostomy and epididymovasostomy for seminal tract obstruction: cause of obstruction and operative outcome]. AB - The surgical outcome in 42 patients with seminal tract obstruction who underwent microsurgical vasovasostomy or epididymovasostomy and were followed up for more than 3 months postoperatively is reported. Causes of the obstruction were vasectomy in 7 patients, childhood inguinal herniorrhaphy in 15, epididymitis in 5, Young's syndrome in 3 and others in 12. Twenty-eight patients had bilateral obstruction while 14 had unilateral obstruction. After 51 operations, including 11 reoperation in 9 patients, the anastomosis was patent in 83.3%, semen quality normalized in 21.4% and pregnancy was achieved in 31.0% of the 42 patients. Success rate of vasovasostomy and epididymovasostomy was 57.7% and 78.3%, respectively. Factors detrimentally affecting the operative outcome were obstruction due to Young's syndrome, obstruction for more than 10 years, absence of sperm in vasal fluid during the operation, elevated serum FSH levels and presence of seminal tract anomaly. Presence of serum antisperm antibodies had no effect on pregnancy rate. Microsurgical reanastomosis of the seminal tract resulted in restoration of fertility in many patients with seminal tract obstruction who had normal testicular spermatogenesis. PMID- 8411823 TI - [Laparoscopic adrenalectomy]. AB - Since January, 1992, 8 patients with adrenal disease have been treated by laparoscopic surgery. There were 4 males and 4 females (35-65 years old), seven patients had primary aldosteronism and one non-functioning adrenal tumor. Six trocar-sheath units were placed in the upper abdomen. In case of the lesion being on the right side, the posterior layer of the peritoneum was incised at the point of the hepatic flexure of the colon and on the left side, the dissection was begun by dividing the peritoneal reflection over the lateral aspect of the descending colon, adrenal tumors were removed with normal adrenal gland. In all cases adrenal tumors were removed successfully. The operative time was from 165 to 572 (293 min., average) minutes and there was no major complication. PMID- 8411824 TI - [The local relapse of prostatic cancer and initial local treatment]. AB - We treated fifty cases of prostatic cancer that had no distant metastasis and no other primary cancers between October 1983 and September 1990. Our initial treatment for clinical stages A2, B1, B2 included radical prostatectomy, pelvic lymphadenectomy, and bilateral orchiectomy. Staging lymphadenectomy with bilateral orchiectomy was applied to patients with clinical stage C tumor, and if there was no lymph node metastasis, external beam radiation therapy for the prostate was performed. Patients who did not consent to major surgeries or had poor general status or had clinical stage D1 tumor underwent bilateral orchiectomy. Mean observation period was forty-one months. Eleven cases relapsed. Of the eleven cases, ten had local relapse at the diagnosis of relapse (two of them had distant metastasis simultaneously, six of them had distant metastasis later, and two of them had no distant metastasis in the observation period). All these ten cases had no underwent the initial local treatment. The remainder who had underwent radical prostatectomy relapsed with distant metastasis alone. This study suggests that the local relapse tends to precede distant metastasis as the mate of relapse of prostatic cancer excluding stage D2. There is a possibility that initial local treatment is useful improving the prognosis of the patients for clinical stage A2-D1 prostatic cancer. PMID- 8411825 TI - [A case of penile revascularization by Hauri's method for arteriogenic impotence]. AB - In a 35 year old arteriogenic impotent patient without a history of hypertension, arteriosclerotic disease, or diabetes mellitus, the corpus cavernosum of the penis was revascularized using Hauri's method. Before surgery, erection after the intra-cavernous injection of 20 micrograms of prostaglandin E1 was very weak. In a color ultrasonography the peak systolic velocity of the cavernous arteries was recorded as being only 18 cm/sec. Furthermore, no artery except the right dorsal artery was evident even with a digital subtraction angiography. Accordingly he was diagnosed as having arteriogenic impotence, and we carried out the corpus cavernosum revascularization using Hauri's method under microscopic magnification. The dorsal artery and the deep dorsal vein were anastomosed side to-side, and the hypogastric artery and dorsal artery were anastomosed end-to side. After the revascularization surgery, the peak systolic velocity of cavernous arteries returned to normal (53 cm/sec), and the penis showed complete erection after an intracavernous injection of 20 micrograms of prostaglandin E1. Before surgery this patient had no experienced sexual intercourse, but he could achieve full sexual intercourse 2 weeks after the surgery. His erectile ability has been maintained for 4 months since the surgery. This is the 1st case of arteriogenic impotence treated using Hauri's method in Japan. PMID- 8411827 TI - Membership directory 1993. PMID- 8411826 TI - [Renal metastasis from osteogenic sarcoma: report of a case]. AB - Renal metastases of osteogenic sarcoma are unusual clinical occurrence and cases of osteogenic sarcoma treated by nephrectomy for renal metastases have been rarely reported. We report a 19-year-old male who was successfully treated by left nephrectomy for a metastatic tumor of osteogenic sarcoma from the right femur. In 1989 above-knee amputation was performed and the initial relapse with right pulmonary metastasis was treated by right upper lobectomy in 1991. In February 1992 he developed left renal and left pulmonary metastasis, hence left nephrectomy and left upper lobectomy were performed. Metastases of osteogenic sarcoma to the organs other than the lung and bone will increase because of the improvement of diagnostic modalities. Our study suggests that a bone scan is useful for routine follow-up of patients with osteogenic sarcoma and that resection of the metastatic lesion can improve these patients' quality of life. PMID- 8411828 TI - A rule of our own. PMID- 8411829 TI - [Retrograde conduction (ventriculo-atrial) in patients with AV or intraventricular block. Electrophysiologic evaluation]. AB - The purpose of the study was to evaluate ventriculoatrial conduction in patients with chronic complete AV block (AVB) (35 pts--gr. A), who were qualified for pacemaker implantation and in patients with bifascicular block (BFB) (20 pts--gr. B), who had electrophysiological study performed to determine indications for permanent pacing. Anterograde and retrograde conduction by accessory pathways were excluded in all patients. Incremental ventricular pacing (60-80 bpm to 200 220 bpm) and programmed ventricular stimulation were performed. Ventriculoatrial (VA) conduction was present in 12 of 35 patients in gr. A (34%) and in 15/20 patients in gr. B (75%). Mean VA conduction time was 192 +/- 50 ms in gr. A and 200 +/- 54 ms in gr. B, effective refractory period of right ventricle (ERP-RV) was 270 +/- 30 ms and 250 +/- 29 ms respectively. Mean retrograde Wenckebach point was 128 +/- 41 bpm in gr. A and 132 +/- 30 bpm in gr. B. The level of VA Wenckebach block in gr. A was found at AVN in 4 of 12 patients and in remaining was not directly recognised. Levels in gr. B were: AVN--6, not recognised--9 patients. Ventriculoatrial conduction shows complex properties and dependence on the type of ventricular stimulation. Electrophysiological study enables to find a level of retrograde block in some patients. PMID- 8411830 TI - [Left ventricular hypertrophy and blood pressure values during 24-hour ambulatory monitoring and office measurements in essential hypertension]. AB - 119 hypertensive patients were examined by 24-hour ambulatory blood pressure monitoring (ABPM) and traditional (casual) blood pressure measurements. The echocardiographic examination was also performed. Mean and maximal systolic and diastolic blood pressure values, systolic and diastolic pressure amplitude as well as traditional blood pressure values were correlated with left ventricular walls thickness and left ventricular mass. Maximal values were obtained for left ventricular mass and mean systolic blood pressure (especially in the evening). No statistically significant correlation was stated for left ventricular mass and systolic pressure amplitude. The correlation coefficients of left ventricular mass and wall thickness with ABPM were significantly higher than those calculated from echocardiographic parameters and traditional blood pressure measurements. PMID- 8411831 TI - [Magnesium, potassium and sodium in serum and erythrocytes in acute myocardial infarction]. AB - Serum concentration and erythrocyte contents of Mg++, K+ and Na+ was measured on day 1, 3 and 14 in patients with transmural myocardial infarction. The study group consisted of 72 patients (15 females and 57 males) with the diagnosis of myocardial infarction based on WHO criteria. The control group consisted of 52 healthy subjects (11 females and 41 males). Measurements of sodium and potassium were made using flame photometer, magnesium was measured with atom adsorption spectrophotometry. Lowest magnesium and potassium concentrations (in serum and in erythrocytes) were found on day 1 and day 3 with almost full normalization on day 14. Sodium serum concentration and erythrocyte contents varied; lowest serum concentration appeared on day 1 and day 3, erythrocyte contents was highest on day 1. PMID- 8411832 TI - [Chronic thromboembolic pulmonary hypertension--observation of personal case reports]. AB - The clinical picture, differential diagnosis and prognosis in chronic thromboembolic pulmonary hypertension (CTEPH) is being discussed on the basis of a dozen of the authors' cases. The clinical course of CTEPH in 2 patients fulfilling the main criterium for pulmonary thromboendarterectomy and undergoing one year anticoagulation is examined. As a result patient with one large pulmonary vessel occlusion (a. lobaris superior sinistra) improved his clinical status as well as hemodynamic and echocardiographic outcome. In patient with multivessel pulmonary occlusion gradual clinical aggravation and hemodynamic echocardiographic progression of CTEPH was observed. PMID- 8411833 TI - [Recurrent attacks of antidromic tachycardia in a patient with WPW syndrome and latent accessory pathway A-V conduction]. AB - A case of a 38 year old male with WPW syndrome and frequently recurrent antidromic atrio-ventricular sustained tachycardias is presented. In his ecg recordings delta wave depolarizations indicated right free wall accessory pathway. This was proved during electrophysiologic study. The effective refractory period of this pathway was short in both directions-below 200 ms. There was a wide zone of sustained antidromic tachycardia 236-247/min induction. Intravenous injection of flecainide terminated antidromic tachycardia but a wide zone of sustained orthodromic tachycardia 150/min induction appeared. Dissection of the right free wall accessory pathway was performed using Sealy technique (Cox modification). During the intraoperation electrophysiologic study an orthodromic tachycardia with retrograde conduction through an antero-septal accessory pathway was induced. After dissection of that second pathway abnormal atrioventricular conduction was abolished. The patients has been free from episodes of tachycardia during one year follow-up period. PMID- 8411834 TI - [Lipoprotein (A). Missing link in pathogenesis of atherosclerosis?]. PMID- 8411835 TI - [Contemporary clinical possibilities for dynamic applications of cardio myoplasty]. PMID- 8411836 TI - [Serious ventricular arrhythmias in patients without structural heart disease. Results of electrophysiologic testing]. AB - Invasive electrophysiological testing with programmed electrical stimulation (PES) of the heart is widely used to reproduce spontaneous ventricular tachycardia (VT). In patients with coronary heart disease it has been demonstrated, that induction of sustained monomorphic VT (SMVT) was highly predictive of an increased risk of sudden death. The value of the results of PES in patients with idiopathic VT is still controversial. In this study clinical and electrophysiological data were analyzed for 34 patients with apparently normal heart and serious ventricular arrhythmias: 17 of them (group A) had documented VT (n = 9) or VT was suspected on the basis of the presence of unexplained syncope and ventricular arrhythmias in Holter monitoring (n = 8). The other causes of syncope (carotic sinus syndrome, sick sinus syndrome, atrio-ventricular block, neurological causes) were excluded in these patients. The remaining 17 patients suffered only palpitations (group B): in 5 of them Lown's grade 4 and in 12 Lown's grade 1 or 2 ventricular arrhythmia was recorded in Holter monitoring. In both groups coronary heart disease, congenital or acquired valvular heart disease, cardiomyopathy and myocarditis were excluded by means of noninvasive (physical examination, X-ray, ultrasonocardiography, electrocardiographical stress testing) and, when needed, invasive (selective coronarography, endomyocardial biopsy) evaluations. There was no significant differences in age and sex between the groups. All patients underwent baseline drug-free electrophysiologic studies with programmed right ventricular stimulation. The end point of stimulation was the induction of SMVT.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411837 TI - [Relation between erythropoietin production and plasma renin activity in patients with essential hypertension]. AB - Regulation of renal erythropoietin (EPO) production has not yet been clearly established in physiologic as well as in pathologic conditions. Recent studies suggest a possible role for renin-angiotensin system in control of EPO production. An elevation in blood pressure occurs commonly in patients with various forms of anaemia treated with recombinant human EPO. Furthermore it has been found that EPO can alter secretion of hormones engaged in regulation of intravascular fluid volume and vascular resistance. The aim of this study was to determine whether patients with essential hypertension (EH) and healthy subjects differ in EPO secretion and whether EPO serum level is related to renin response to dietary sodium restriction and upright position of the body. 63 patients with EH and 12 healthy subjects were investigated. Patients with EH were divided into subgroups on the following criteria: renin response to dietary sodium restriction and upright position of the body. 63 patients with EH and 12 healthy subjects were investigated. Patients with EH were divided into subgroups on the following criteria: renin response to dietary sodium restriction and upright position of the body and severity of existing hypertension. In all subjects haematocrit value, haemoglobin concentration, erythrocyte count, sodium, potassium, creatinine, iron, ferritin serum levels, total iron binding capacity, plasma renin activity (PRA), erythropoietin serum level and mean arterial blood pressure (MAP) were measured in basic conditions (normal sodium diet). Additionally PRA, EPO and MAP were measured after dietary sodium restriction for three days and upright position of the body for three hours.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411838 TI - [Circadian rhythm of blood pressure and heart rate in hypertension with type 2 diabetes mellitus]. AB - Hypertension is an important risk factor for cardiovascular complications of diabetes. Most of the studies performed in diabetics so far, however, have dealt with the assessment of blood pressure by traditional sphygmomanometry. In order to investigate the circadian pattern of blood pressure and heart rate in patients with different categories of glucose tolerance, we performed ambulatory blood pressure monitoring in 28 obese hypertensives without clinical nephropathy divided in two groups. Group A consisted of 14 hypertensive males with type 2 diabetes mellitus (mean age 49.7 +/- 7.1 years, mean duration of diabetes 4.0 +/- 2.9 years); group B consisted of 14 hypertensive males with normal glucose tolerance according to National Diabetes Data Group (mean age 47.2 +/- 7.1 years). There was no significant difference in casual blood pressure (151.4/104.8 in group A versus 148.5/104.2 mmHg in group B). Ambulatory blood pressure monitoring revealed significantly higher systolic blood pressure in group A during the day (162.2 +/- 12.1 vs 152.1 +/- 9.0 mmHg in group B, P < 0.05) and at night (141.0 +/- 13.2 vs 125.5 +/- 12.5 mmHg in group B, P < 0.005). That suggests that casual readings underestimate systolic blood pressure as a predictor for macrovascular events in type 2 diabetics. The decline in nocturnal heart rate was significantly lower in group A (11.2 +/- 5.2 min-1) in comparison to group B (16.9 +/- 7.0 min-1; P < 0.05) suggesting reduced parasympathetic tone at night in diabetic patients. We conclude that type 2 diabetes has significant influence on systolic blood pressure and heart rate 24-h profiles even in patients without diabetic nephropathy. PMID- 8411839 TI - [Atrial septal aneurysm complicated by mitral valve prolapse--two cases]. AB - Atrial septal aneurysm is a rare heart defect. We decided to present two cases of this defect complicated by mitral valve prolapse. The diagnosis was established by transthoracic 2D-echocardiography. Remembering that atrial septal aneurysm often co-exists with atrial septal defect we performed contrast echocardiography. The examination did not show any shunts between atria. Ventricular ectopic beats were observed in both cases. Cardiac arrhythmia was successfully treated with B blockers. PMID- 8411840 TI - [Interaction of drugs used in cardiology]. PMID- 8411841 TI - [Paroxysmal loss of consciousness. Comparison of electrophysiologic and holter monitoring results]. AB - Holter monitoring is one of the most commonly done test in the evaluation of patients with syncope. As Holter monitoring may miss an arrhythmia or document an asymptomatic arrhythmia invasive electrophysiologic testing has been applied in patients with syncope. The purpose of this study was to compare Holter monitoring and electrophysiologic tests in patients with syncope of unexplained origin. The group consisted of 100 patients with syncope of unknown origin. Coronary artery disease was present in 43 patients, other heart disease in 19 patients and no structural heart disease was found in 38 patients. Electrophysiological testing consisted of (1) recording of His bundle electrogram, (2) atrial stimulation, (3) ventricular stimulation with 1.2 and 3 extrastimuli at three basic cycle lengths. The results of Holter monitoring were classified by severity of abnormalities into three classes: I--normal study; II--moderate abnormalities; III--severe abnormalities: sinus rhythm with pauses longer than 3 s, Mobitz II or complete atrio-ventricular block, supraventricular arrhythmia faster than 180 bpm, sustained ventricular tachycardia. Abnormalities of electrophysiologic testing were grouped as: I normal study; II--moderate abnormalities; III--severe abnormalities: sinus nodal recovery time more than 3 s, HV interval longer than 100 ms, supraventricular arrhythmia faster than 200 bpm, sustained ventricular tachycardia. Class III abnormalities were documented in 17 patients on Holter monitoring and in 20 patients by electrophysiologic testing. Compatibility between class III abnormalities in Holter monitoring and electrophysiological testing was noted in 4 patients, discordance of class III results in 33 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411842 TI - [Accelerated coronary angiopathy in patients after cardiac transplantation]. PMID- 8411843 TI - [Hemophilus influenzae infection and their prevention by vaccination]. AB - Encapsulated and non-encapsulated strains of haemophilus influenzae are known to exist. The encapsulated ones, especially those of type B (HiB), are highly invasive. Hib may cause, among other diseases, purulent meningitis, epiglottitis, septic arthritis/osteomyelitis and pneumonia. These diseases can be prevented by timely vaccination. The non-encapsulated haemophilus ionfluenzae strains often produce colonisation of the nasopharyngeal space and inflammation of the mucosa of the airways. They are often pathogens of otitis media, sinusitis, bronchitis and pneumonia. These diseases cannot be prevented by HiB vaccination. PMID- 8411844 TI - [Differential diagnosis and therapy of hypoglycemia in childhood]. AB - Occurrence of hypoglycaemia is directly connected to fuel balance. Compared to free fatty acids, glucose is rapidly exhausted. Therefore, besides disturbances in available glucose (i.e. defects in gluconeogenesis or glycogenolysis) as well as their hormonal control, defects of beta-oxidation might be possible causes. According to recent investigations they occurred with similar frequency. A blood glucose level of 2.6 mmol/l (= 47 mg%) already represents a critical concentration of brain function. Based on pathophysiological concepts concentrations of glucose, free fatty acids, ketone bodies and insulin must be determined from a single plasma sample. Together with case history and clinical symptom findings, these parameters are useful to get a reliable tentative diagnosis. Therefore, descriptions like "ketotic" or "idiopathic" hypoglycaemia can be avoided and suitable treatment can be started. Suspected diagnosis must be confirmed by means of chromatographic, enzymatic, and if necessary histological investigations. Successful treatment depends mainly on early diagnosis. This is of importance in case of surgery in organic hyperinsulinism, but likewise for advisory help to families with infants suffering from beta-oxidation defects, to avoid prolonged fasting. PMID- 8411845 TI - [Perinatal herpes infection. Clinical aspects--therapy--follow-up]. AB - Herpes simplex virus may cause serious infections in neonates. In case of foetal infection in the first trimenon, abortions, stillbirth, prematurity, intrauterine growth retardation (not obligatory) and various malformations may result. Neonatal HSV infection is mostly the consequence of intrapartum virus acquisition during passage through the birth canal. The infection is mostly localised on the skin, at the eyes or the mouth or disseminated with or without HSV meningoencephalitis. It is difficult to establish the diagnosis, because neonatal herpes disease in the early stage is not easy to distinguish from other diseases in the newborn such as RDS, NEC or ICH. Antiviral therapy with aciclovir is the treatment of choice and seems to improve the outcome of neonatal herpes. Prognosis depends on early therapy. Treatment should be initiated in relation to clinical findings, because available diagnostic techniques do not always permit an early detection of the disease. PMID- 8411846 TI - [Compliance of children with bronchial asthma in long-term therapy with theophylline retard preparations]. AB - 427 samples of the saliva of 235 children were taken on an outpatient basis to follow up long-term theophylline treatment. 37 samples from 34 children did not reveal the presence of any theophylline. The absolute non-compliance found in this manner is 21% in the 9 to 12 years age bracket and significantly higher than with the children below 9 years and above 14 years of age (8 and 10%, respectively). Better differentiation of compliance and separation from underdosing was achieved in 38 children by comparing the outpatient-controlled theophylline levels with the values obtained on an inpatient basis. For 10 patients this method definitely revealed non-compliance, whereas compliance was doubtful with 12 children and 15 children had strictly adhered to the prescribed dosage regimen. To improve the intake discipline the theophylline levels should be regularly followed up under outpatient conditions and evaluated in cooperation with the patients. PMID- 8411847 TI - [Oral ciprofloxacin therapy in juvenile patients with cystic fibrosis--results of a prospective pilot study]. AB - Efficacy and safety of oral ciprofloxacin were studied in a prospective study at three cystic fibrosis centres, covering 24 in-patients suffering from cystic fibrosis and acute bronchopulmonary exacerbation. The patients were between 10 and 17 years of age. Pseudomonas infection was present in 75% of these patients. Despite frequent persistence of the pathogens, clinical improvement was noted in 75% of the treated children. A definite increase of the average MIC was not seen in 20 cases of persisting strains. No serious side effects occurred during the 14 day oral treatment course. Ciprofloxacin is a useful alternative to conventional parenteral treatment with antibiotics in patients suffering from cystic fibrosis and infections of the airways. PMID- 8411848 TI - [Intrauterine stab injury with a knife in the head of a fetus in the 29th week of pregnancy]. AB - Penetrating head trauma in children causes uncommon and potentially life threatening injuries. We report on a case of penetrating cranial stab wound to the right parietal region of the head to a 29-week fetus. The child was delivered by emergency Caesarian section. Neurosurgical intervention after birth was necessary. The injury caused a posthaemorrhagic hydrocephalus. Real-time ultrasound examination visualises the brain damage and the development of a hydrocephalus e vacuo. PMID- 8411849 TI - [Experience with the glycerol lysis test in acid medium in diagnosis of hereditary spherocytosis]. AB - The acidified glycerol lysis test (AGLT) is highly sensitive for hereditary spherocytosis (HS). In all 50 patients of the Children's Hospital of the Medical Academy Magdeburg suffering from HS the 50% lysis time was found to be pathological between 19 and 110 seconds. However, pathological results of this test were also found in autoimmune haemolytic anaemias. AGLT was negative in non spherocytic haemolytic anaemias and normal controls. In conclusion, the AGLT is a rapid, simple and inexpensive screening procedure for spherocytes in blood. PMID- 8411850 TI - [Therapy of idiopathic thrombocytopenic purpura in childhood]. AB - Idiopathic thrombocytopenic purpura characterised by an increased bleeding tendency is a well-known clinical entity in childhood. From 1983 to 1992 68 patients suffering from ITP were treated at the Children's Hospital of the Medical Academy of Magdeburg. 11 patients with mild or without clinical symptoms and platelet counts of more than 20 Gpt/l did not receive any treatment; all children recovered spontaneously. 38 patients with severe haemorrhagic manifestations and thrombocytes less than 20 Gpt/l were treated with corticosteroids and had a sustained remission. 10 patients who had responded to corticosteroids initially and subsequently relapsed were given other treatments (Anti-Rhesus-antibodies, HDIVG). All patients achieved a continuous remission. A chronic disease was observed in 8 patients; 3 of them were splenectomised. One child died due to massive gastrointestinal bleeding. PMID- 8411851 TI - [BNS epilepsy after the second triple (DPT) vaccination?]. PMID- 8411852 TI - [Extremely marked local reaction (erythema, swelling, pain) after the 1st and 2nd DPT vaccination?]. PMID- 8411853 TI - [The surgery of huge retroperitoneal tumors]. AB - Observations of successfully operated huge retroperitoneal tumors, one of them weighing 14.700 kg, are reported. The advantages for precise diagnosis of modern visualizing techniques--echography and computer tomography--and the inability to establish always an exact cytologic diagnosis by puncture aspiration biopsy are demonstrated. Total extirpation of the tumor with subsequent combined chemo-, radio- and immunotherapy is advised. PMID- 8411854 TI - [Hemorrhage from the digestive tract in the Mallory-Weiss syndrome]. AB - Thirty-three patients with Mallory-Weiss syndrome have been treated at the Clinic of Emergency Surgery in Sofia for a period of 11 years (1980-1990). According to treatment and outcome they were classified as follows: conservative treatment--7 patients, electrocoagulation--16, sclerosing procedure--10, successful endoscopic hemostasis--17, emergency surgical intervention--9, relapse of bleeding after endoscopic surgery--9, death from hemorrhage--4, total case fatality--12.1 per cent, operative case fatality rate--33.3 per cent. PMID- 8411855 TI - [The modern aspects of the surgical treatment of the perforated gastric and duodenal ulcer with a view to the immediate results]. AB - Experience is recorded with the operative treatment of 185 patients with perforated gastric and duodenal ulcer over the period 1981-1991. The period was divided in two: 1981-1985 and 1986-1991. During the first period were operated 89 patients: 58 with perforated duodenal ulcer and 31 with gastric ulcer. The method of choice during this period was suture of the perforation. During the second period were operated 96 patients: 90 with duodenal and 6 with gastric ulcer. Surgical approach was also individualized depending on the pathologic process, patient age and accompanying diseases. The following operative interventions were performed: "pure suture" of the ulcer--55 patients; gastric resection by Billroth I and II techniques--7 patients; different variants of vagotomy with drainage operation--34 patients. Analysis of the immediate results in 42 patients demonstrated that best results were obtained by vagotomy + drainage operations. PMID- 8411856 TI - [The immediate results of transendoscopic sclerotherapy in hemorrhaging varices, tumors and polyps and in the Mallory-Weiss syndrome]. AB - In 1982 four Japanese authors Okaba, Honda, Toshiba and Asami reported at the International Symposium in Tokyo on successful transendoscopic sclerosification of bleeding ulcer, around which a sclerosifying agent or absolute alcohol was injected with or without epinephrine. Alcohol suppresses bleeding, causing local dehydration, coagulation and sclerosis of the bleeding tissue, thus producing hemostasis. For the last two years the authors have gained some experience in the application of periulcer and peritumor sclerosification with the sclerosifying agent etoxysclerol (3 per cent solution) and pure 96 per cent ethanol + 1 ampoule epinephrine. As regards the bleeding site, patients with the following nosologic entities were treated: gastric ulcer--24 patients; duodenal ulcer--13; gastric cancer--6; anastomotic ulcer--2; acute ulcer--7; Mallory-Weiss syndrome--10 patients. The results may be summarized as follows: temporary hemostasis was achieved in 21 patients, bleeding recurred in 8 and 9 patients required emergency surgery. Complete and ultimate hemostasis was attained in 29 patients (46.7 per cent). Nine patients died of hemorrhage. PMID- 8411857 TI - [Nalbuphine hydrochloride (Nubain) for postoperative analgesia]. AB - A new opioid analgetic (Nalbufin hydrochloride) was applied to 28 patients having undergone abdominal operations. The time of application of the analgetic agent was determined by the patient himself, in accordance with the five-grade classification of pain, offered by Beaver and Feise. It is pointed out that nalbufin hydrochloride has adequate analgetic power and duration of analgetic activity which makes it suitable for postoperative analgesia. PMID- 8411858 TI - [Hemorrhage from acute erosive processes in the upper digestive tract in thermal trauma]. AB - Forty-six patients with thermic trauma complicated by bleeding from the gastroduodenal tract have been observed during the period 1975-1985. The nosologic entities with bleeding symptom were as follows: hemorrhagic gastritis- 2 patients, acute ulcer--5, inveterate callous ulcer--5. Transendoscopic operation (electrocoagulation + sclerosification) was performed in 5 patients with acute ulcers, and conventional surgery--in patients. Nine patients died, 7 of them operated. Overall case fatality rate was 19.6 per cent, operative case fatality rate--87.5 per cent. Type of surgical intervention: gastric resection by Billroth II technique--5 patients; gastrotomy with suturing of the ulcer--3 patients. PMID- 8411859 TI - [Therapeutic esophagoscopy based on a portal and preportal block in hemorrhaging varices of the esophagus and portal hypertension]. AB - Parallel with the hitherto known methods of treatment of bleeding esophageal varicosities in portal hypertension with Seinkstaken-Blackmore's probe and by surgical approaches for controlling the hemorrhage from ruptured esophageal varicosities, such as Cryle's and Linton's operations, azigoportal deconnection by Tanner's technique, splenectomy, omentohepatoplexy, Sugiura's operation, esophagus ligation by the method of Fosschulte and V. Mateev (1983), the authors apply the method of transendoscopic sclerosification of bleeding varicosities in adults and children with cirrhosis of the liver and prehepatic block. The indications and contraindications of the method and the etiologic factors underlying portal and preportal block in adults and children (in adults- cirrhosis of the liver, in children--mostly preportal block secondary to portal vein thrombosis dating back to early childhood after exchange transfusion through the umbilical vein) are discussed. Of 11 sclerosified children 3 to 14 years of age, 10 are still alive (100 per cent survival of living patients). For the period 1985-1990 a total of 60 adult patients with bleeding ulcers were sclerosified and resclerosified. Hemostasis attempted in 11 patients provided successful obliteration in 42 per cent; endoscopic failure--in 15 patients (57.69 per cent). Seven patients were operated, 5 of them died. Fourty-off of 53 nonoperated patients died. Overall case fatality rate was 78.33 per cent, operative case fatality rate--71.42 per cent. Fifteen of the deceased nonoperated patients died of uncontrollable persistent bleeding, 27 of severe hepatic failure, progressive jaundice, ascites hepatorenal insufficiency and hepatargia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8411860 TI - [Comparative studies of acid-base and gas metabolism in the whole blood and erythrocytes in endotoxic shock in rabbits]. AB - Parameters of acid-base and gas metabolism in whole blood and in red cell hemolysate were determined 30, 60 and 120 minutes after inducing endotoxin shock in rabbits of Belgian giant breed. Astrup's micromethod on apparatus of the firm "Radiometer" was used. Endotoxin shock was induced by intravenous injection of 2 mg/kg endotoxin from E. coli 0111:B4 strain. Equations were used for expressing the relation between pH of erythrocytes and pH of whole blood and analysis made of the correlation between the changes in the individual components of erythrocyte and of whole blood acid-base metabolism. Severe metabolic acidosis developed both in whole arterial blood, in mixed venous blood and in red cell hemolysate. Close correlation was recorded between the changes in the parameters of acid-base metabolism in whole blood and in red cell hemolysate on the 60. and especially on the 120. minute after endotoxin administration. The blood gas changes manifested by slight decrease of PCO2 and PO2 both in arterial and in mixed venous whole blood and red cell hemolysate were not statistically significant. The severe tissue hypoxia during the early phases of endotoxin shock was thought to be the result of severe hemodynamic changes. PMID- 8411861 TI - [The leading symptoms in patients with renal carcinoma]. AB - The incidence of the leading symptoms of renal cancer in 97 patients operated at the University Urologic Clinic in Rostock, Germany, was studied. In all patients the diagnosis was clinched by histologic examination. RESULTS: gross hematuria had 43 per cent of the patients; pain in the lumbar area--29 per cent; arterial hypertension--27 per cent; change finding--21 per cent; conventional tumor symptoms--17 per cent; palpable tumor--11 per cent; classical triad--8 per cent. The incidence of the conventional tumor symptoms was: loss of weight--17 per cent; fever--21 per cent; malaise--17 per cent. The results of this study showed that gross hematuria was the most common symptom. These results as a whole considerably differ from data reported by other authors. Common to all is however the top incidence of gross hematuria and pain in the lumbar area. PMID- 8411862 TI - [The incidence, localization, sex and mean age in patients with renal carcinoma]. AB - Retrospective analysis was made of the clinical case records of 97 nephrectomized patients with renal cancer at the University Urologic Clinic in Rostock, Germany. The Student-Fisher t-criterion was used for statistical processing of the results. The stage of the disease was determined in accordance with the histologic classification of pTNM, suggested in 1988 by the European Organization for Cancer Research and Treatment. Patients with malignant diseases of the urogenital tract accounted for 6.1 per cent of all patients treated at the Urologic Clinic. Malignant disease of the kidney alone comprised 1.7 per cent and of renal cancer--1.5 per cent. In 49 per cent of the patients renal cancer was localized in the left kidney, in 42 per cent--the right kidney, and 9 per cent both kidneys were involved. The male: female ratio was 2.2:1. Mean patient age was 57 years (males 58.1, females 55.9 years). PMID- 8411863 TI - [The basic principles of diagnosis and treatment in emergency surgery]. AB - A review of the literature is made, in the attempt to establish the principles of the diagnosis and treatment in emergency surgery. They are listed as follows: 1. To preserve patient life and evaluate the functional capacity of the organism; 2. To establish emergency diagnosis; 3. To choose the method of treatment; 4. Method of operation. The guidelines in emergency surgery is to develop and perfect new methods of operation and new technical facilities, to raise the professional skill of the surgeon and to improve the organization of work in surgical clinics and departments. PMID- 8411864 TI - [A case of hypernephroma in a horseshoe kidney]. PMID- 8411865 TI - [A case of carcinoid tumor of the breast]. AB - A rare tumor of the mammary gland, built up of argyrophilic cells, with numerous metastases in the axillary lymph nodes is reported. Histochemical and electron microscopic studied unequivocally demonstrated that this tumor belonged to the APUD system. The possible histogenesis of carcinoid tumor of the mammary gland is briefly discussed. PMID- 8411866 TI - [A case of epidermoid cysts in the spleen]. AB - A rare case of epidermoid cysts of the spleen in a 16-year-old boy, presenting with early clinical symptoms, is reported. In view of the extreme rarity of primary cysts of the spleen, the authors contemplate on the possibilities for establishing the diagnosis. PMID- 8411867 TI - [A method for laparotomy in appendectomy]. PMID- 8411868 TI - [A choledochal elevator]. PMID- 8411869 TI - [The clinical use of lincomycin in treating anaerobic infection in patients with surgical abdomen]. AB - Experience is recorded with lincomycin treatment of 59 patients with acute surgical abdomen. The primary anaerobic flora was eradicated in 86 per cent of the patients. For the very good therapeutic results essential was the early instituted adequate antibiotic therapy in adequate dose and for adequate length of time. PMID- 8411870 TI - [Changes in the cell cycle of cultured lymphocytes from patients subjected to halothane anesthesia]. AB - The effect of halothane anesthesia on the proliferative activity of peripheral blood lymphocytes was analyzed. Included in the study were 7 patients. Lymphocyte cultures were prepared by standard methods with blood samples obtained immediately before the operation, after the effect of halothane anesthesia was over and 24 hours later. 5-bromodeoxyuridine in concentration 10 micrograms/ml was used for labelling cells which had passed 1, 2 or more mitotic cycles within the cultivation time. Cell proliferation rate was evaluated by the proportion of metaphases from I, II and III mitotic cycle. Lymphocyte proliferation in the cultures was shown to be inhibited after halothane anesthesia. Inhibition was particularly strong in the first post-halothane sample, but kept on being strong also 24 hours later. The first mitotic cycle was markedly prolonged--to such a degree that subsequent cycles within the 72-hour cultivation period were fully absent. The results were discussed in the light of the overall effect of halothane in the process of its metabolism, its effect on DNA synthesis and eventual implication of additional immunologic mechanisms. PMID- 8411871 TI - [The clastogenic effect of halothane on the lymphocytes from patients operated on under halothane anesthesia]. AB - The clastogenic effect of halothane anesthesia on peripheral blood lymphocytes from operated patients was studied. Chromosomal aberrations and sister chromatic exchanges in lymphocyte cultures of 7 patients were analyzed. Blood samples were obtained before the operative intervention, immediately after the effect of anesthesia was over and 24 hours later. To differentiate sister chromatids in the culture medium, on the 24th cultivation hour bromodeoxyuridine in final concentration 10 micrograms/ml was added. Increased incidence of structural chromosomal aberrations was recorded in lymphocyte cultures of patients after anesthesia--6.43 per cent cells with aberrations in the first post-halothane culture and 8.28 per cent in the second. Comparison of the cell affection index with the total number of affected cells revealed a continuous clastogenic effect pattern within the study period. Reliable evidence for induction of sister chromatid exchanges after the anesthesia was not found. The results were debated in the light of the possible mechanisms of clastogenicity of halothane anesthesia, the initial state of the patients and the consequences for them. PMID- 8411872 TI - [Bronchogenic cysts of the mediastinum in childhood]. AB - Bronchogenic cysts are rare congenital anomalies result of abnormal development of the tracheobronchial tree. They develop in the lung parenchyma or in the mediastinum. For a period of 17 years (1975-1991) a total of 46 children with bronchogenic cysts have been treated; in 6 the cysts were localized in the mediastinum. Two patients were one-year-old, 3 were in the age group 1 to 3 years and 1 was older than 3 years. Four patients were boys and 2 girls. Two children had hacking cough and attacks of dyspnea; one had swallowing difficulties. In 3 children the disease produced no symptoms and was detected on examination for pneumonia (1 child) and acute upper respiratory tract infection (2 children). All children had conventional X-ray; 5 had esophagography as well, 4--computer axial tomography, 3--echography. All children were operated (total extirpation of the cyst) and left the hospital cured. PMID- 8411873 TI - [The effect of nalbuphine hydrochloride (Nubain) on respiration in surgical patients]. AB - The effect of nalbufin hydrochloride (Nubain) on breathing was studied in 28 patients after abdominal operations. It was shown that, regardless of its partial effect on the mu-receptors, Nubain caused no respiratory disturbances. The authors recommend it for analgesia in abdominal operations, but only after careful evaluation of the general status and the respiratory function in the operated patient. PMID- 8411874 TI - [Our experience with diathermy coagulation as a form of therapeutic endoscopy in acute upper digestive tract hemorrhages]. AB - In addition to sclerosifying by use of fibroendoscope which acts upon the source of bleeding, to achieve hemostasis and prevent bleeding relapses nowadays other methods have also gained acceptance. Major importance among these is attached to diathermocoagulation. It may principally be assumed that diathermocoagulation is indicated for critically ill patients with serious accompanying diseases of primary or secondary importance, as well as in the event of persistent bleeding during the clinical examination and impending resumption of bleeding soon after the examination. Hemostasis by electrocoagulation should not be attempted in patients in agonal state when endoscopic examination is in its self dangerous. It is contraindicated also when the source of bleeding can not be established, in cases of severe arterial bleeding, blurring the optics, and in severe concave ulcers carrying the risk of perforation. The method was applied on 173 patients; in 96 (55 per cent) electrocoagulation was successful and in 78 (45 per cent) was unsuccessful. Electrocoagulation was considered a success in patients with acute ulcers and cardiovascular disease, in cases of large erosive units of drug origin, in posteriorly located gastric ulcers, in Mallory-Weiss syndrome and in gastric cancer. PMID- 8411875 TI - [The results and a discussion of the esophagogastroduodenoscopic studies performed successfully, early and postponed in patients with upper digestive tract hemorrhages in 1980-1990]. AB - For a period of 10 years (1980-1990) a total of 2034 emergency, early and postponed endoscopies have been performed in patients with upper digestive tract bleeding at the Department of Emergency Surgery in Sofia. Duodenal ulcer has been the leading cause of bleeding--41.19 per cent of the cases, with gastric ulcer ranking second--14.94 per cent. There followed in descending rank order: gastric and duodenal erosions--12.34 per cent; gastric cancer--5.06 per cent; hemorrhagic gastritis--2.56 per cent; esophageal varices--7.27 per cent; Mallory-Weiss syndrome--1.62 per cent. Bleeding from the digestive tract of patients with CNS trauma based on acute stress ulcer and erosions and activated old callous ulcer occupied the 12th place in rank order. On the 13th place ranked upper digestive tract bleeding in patients who had undergone thermic trauma (2.01 per cent). Much fewer wer the cases of bleeding secondary to peptic ulcer of the jejunum, gastric and duodenal diverticulosis, acute ulcers in patients with cardiovascular diseases, blood diseases, liver, bile duct and pancreas diseases, aorto-duodenal fistulas and drug-induced diseases. The cause remained unknown in 1.77 per cent of the patients. On the basis of the indisputable achievements in [correction of ti] the diagnosis of acute upper digestive tract hemorrhage, the approach to these grave nosologic entities has essentially been altered. PMID- 8411876 TI - [The modern diagnosis of stenosis and obstruction of the ureter]. AB - Included in the study were 681 patients with stenosis and obstruction of the ureter (SOU), diagnosed at the Department of Urology, Clinical Center of Urology, University Aleksandrovska Hospital in Sofia for a period of 5 years. The diagnosis rested on clinical, laboratory, X-ray, radioisotopic, echographic, endoscopic and other investigations. Analysis of the clinical and laboratory data demonstrated an outstanding triad of clinical symptoms: pain (76.8 per cent), hematuria (74.0 per cent), dysuria (70.9 per cent). The laboratory data confirmed the standpoint of other authors that most common is the urinary tract infection, followed in incidence by microhematuria, leukocyturia and hemoglobin content lower than 10 mg %. The most common causative agents of infection were E. coli (29.8 per cent) and Proteus (26.28 per cent). The X-ray methods being used were excretory urography (76.8 per cent) and plain X-ray on kidney-ureter-bladder film (93.2 per cent of the patients). Excretory urography furnished information on the cause, degree and location of SOU and on the effect upon the kidney. It helped to determine the approach to treatment--conservative transureteral or operative. PMID- 8411877 TI - [The modern treatment of stenosis and obstruction of the ureter]. AB - The methods used for treatment of stenosis and obstruction of the ureter (SOU) are: conservative treatment, extracorporal lithotripsy, transureteral operations, surgical management. The different genesis of the nosologic entities underlying- SOU requires adequate effective and organ preserving treatment. In the authors series of 681 patients the mot common approach to treatment und transureteral operations (46.7 per cent) with surgical treatment ranking next (44.5 per cent), presented by organ preserving, organ removing, palliative and reconstructive procedures. Third in incidence was extracorporal lithotripsy of ureteral calculi (31.3 per cent). Judirrdual conservative treatment was least common (7.6 per cent). As a result of treatment in 76 per cent of the patients with "mute kidney" renal and ureteral function were re-established after removal of the stenosis and obstruction of the ureter. PMID- 8411878 TI - [The diseases causing stenosis and obstruction of the ureter]. AB - A total of 681 patients with stenosis and obstruction of the ureter (SOU) have been observed over a period of 5 years. The male/female ratio was 1:1.25. The etiologic factors for development of SOU have been as follows: calculus in the ureter--78 per cent of the total number of patients; infections of the ureter- 8.1 per cent; posttraumatic injury to the ureter--6.5 per cent; congenital malformations of the ureter--4.2 per cent; papillary tumors of the ureter--2.7 per cent; retroperitoneal tumor--0.3 per cent. PMID- 8411879 TI - [Survival and prognosis in renal carcinoma]. AB - A study of the survival and prognosis of renal cancer was performed in 97 patients in different stage of the disease. Mean survival was 2.9 years for males and 3.1 years for females (no statistical significance as to sex). It was shown that with advancing stage of the disease decreased patient survival (T-2 = 4.1 yrs., T-3A = 2.5 yrs., T-3B = 1.3 yrs). The same tendency was observed to a higher degree in N-stage, according to the TNM classification: no single patient survived in N-2 stage beyond 3 years. Observations in M-2 stage showed a much shorter survival time: there was no survival beyond 1 year in M-2 stage. PMID- 8411880 TI - [The surgical treatment of patients with renal carcinoma]. AB - A retrospective study of 118 patients with renal cancer, treated at the University Urologic Clinic in Rostock, Germany, for the period Jan. 1980-March 1990 was performed. Twenty-one patients (17.8 per cent) were in advanced stage of the disease which made operative treatment impossible. Radical nephrectomy was performed in 83 patients: transperitoneal with pararectal approach in 79 and lumbotomy in 4. Partial renal resection was performed in 7 patients, in 5 of them with local hypothermia. The operation in 7 patients terminated with probatory laparotomy because of infiltrative growth of the tumor in adjacent tissues, v. cava included. Probatory laparotomy served for intraoperative biopsy, allowing to establish histologic diagnosis. Six patients died within a month after the operation (6.15 per cent postoperative lethality). Intraoperative complications developed in 4 patients: rupture of the spleen in 2 requiring splenectomy, lesion of colon descendens which necessitated transversectomy--in 1 patient, and rupture of the aorta in 1 patient successfully controlled with two-layer suture of the aortic wall. PMID- 8411881 TI - [Acute rectal hemorrhage]. AB - Problems of the diagnosis, tactics and treatment of acute rectal hemorrhage are discussed. A diagnostic-tactical plan for determining the cause, source, localization and degree of bleeding are discussed. A diagnostic-tactical plan for determining the cause, source, localization and degree of bleeding, the acquired temporary or definitive hemostasis and the degree of blood loss is presented. Adequate approach to acute rectal hemorrhage helps to shorten the operation time, to reduce the extent of preoperative resuscitation and decrease total and operative lethality. PMID- 8411882 TI - [A case of a retrorectal chordoma simulating colonic carcinoma]. PMID- 8411883 TI - [A case of melanoma of the rectum]. PMID- 8411884 TI - [A case of the Zollinger-Ellison syndrome treated surgically]. PMID- 8411885 TI - [An instrument for the mobilization of organs during resection]. PMID- 8411886 TI - [The diagnostic and treatment problems in closed diaphragmatic trauma]. AB - Twenty four patients with blunt trauma of the diaphragm comprise the study group. Four of them died of accompanying injuries in other anatomical regions. All patients had the left leaf of the diaphragm affected. Laceration was most frequently radial in the area of centrum tendineum or in the transition area to pars muscularis. In left-sided ruptures of the diaphragm protruded usually the stomach, left colon, spleen, omentum majus and seldomer small intestines. The clinical picture was multiform and depended mostly on the severity of the accompanying injuries and on the basic complications in ruptured diaphragm- incarceration and strangulation of the protruded abdominal organs. When rupture of the diaphragm is diagnosed operative treatment is as a rule mandatory. The surgical approach to left-sided ruptures of the diaphragm is the abdominal approach, because of the more frequent injury to the abdominal organs. PMID- 8411887 TI - [The current aspects of the diagnosis and procedure in gastroduodenal hemorrhages of ulcerous origin]. AB - Basic problem in all discussions on bleeding gastroduodenal ulcers is the one pertaining to the surgical tactics and to the ascertainment of the indications for surgical intervention and the time for its performance. Adequate answer to this question comes from recognition of the following points: 1. Endoscopically established bleeding source and intensity; 2. Considerations on other endoscopic criteria, clinical manifestations of bleeding--hematemesis, melena or both, changes in the controlled hemodynamic and hematologic parameters, as well evaluation of the accompanying diseases of major and secondary importance on the part of the cardiovascular and respiratory system; 3. Site and time of application of electrocoagulation and periulcer sclerosification in patients considered adequate for them, complying with the indications and contraindications; 4. Adequately chosen operative method. For the period 1985 1990 a total of 81 patients with bleeding from gastroduodenal ulcers, uncontrollable by conservative treatment, have been operated; 37 of them died (overall lethality 13.96 per cent, operative lethality 45.67 per cent). The choice of surgery depended on the localization, nature and extent of the pathologic process, the type and anatomical position of the pathologic focus, correlation with tissues adjacent to the pathologic process, presence of adhesions, callosity, etc. The method of choice was gastric resection by Billroth I, which was preferred to Billroth II resection. Other methods being used were: excision and pyloroplasty, excision, pyloroplasty + vagotomy, suture + vagotomy- mainly for ulcers localized on the anterior duodenal wall, suture, suture + vagotomy, ligation of a. gastrica sin., etc. PMID- 8411888 TI - [Streak retinoscopy. Optical principles and practical recommendations]. AB - The optical principles of streak retinoscopy are demonstrated by diagrams and by optical models. The phenomena observed in the proband's pupil are being explained in two ways: In the first explanation, the image on the proband's retina and its movements are considered, or rather their projection into space; this explanation corresponding to the term "retinoscopy" is commonly used in English textbooks. In the second explanation, the limitations of the light bundle by the proband's and the observer's pupils are considered; hence the term skiascopy (sigma kappa iota alpha = shadow), commonly used in the German literature. The practical hints include retinoscopy through the phoropter, suggestions for the examination of children both with and without cycloplegia, the measurement of accommodation, the detection of opacities and irregularities in the optical media, and the analysis of visual field defects caused by optical anomalies, such as refraction scotomas. PMID- 8411889 TI - [Multifocal intraocular lenses--an assessment of current status]. AB - Besides the diffractive multifocals, which produce a second focus for near vision by means of diffraction rings, there are different refractive multifocal IOL types with 2-7 refractive zones or an aspheric/spherical construction principle. Long-term results: 2 years after implantation of diffractive multifocal IOLs, the corrected distance and near acuities were unchanged compared to the 3-month results. The uncorrected distance acuity was, however, slightly decreased due to a minus shift of refraction to -1.2 D. The contrast sensitivity was improved after 2 years. Multi- versus monofocal IOLs: After diffractive multifocal IOL implantation, the near acuity with distance correction only was markedly improved compared to monofocal IOLs. All other acuity data did not differ between multi- or monofocal lenses. The contrast sensitivity (at low contrasts and high spatial frequencies) and mesopic visual acuity (without and with glare) were reduced compared to monofocal pseudophakic eyes. Near aniseikonia and binocular functions: In unilateral multifocal pseudophakia (monofocal IOL in fellow eye), a near aniseikonia up to 8% was found. The width of fusion was significantly lower than in bilateral multifocal pseudophakia, whereas the stereopsis showed no difference. Determinants of bifocal function: In 7.1% of our cases, no bifocal function (BFF) was present after implantation of diffractive multifocal IOLs. These patients exhibited a significantly higher age as well as higher pre- and postoperative astigmatism, when compared to patients with good BFF. Optical performance of different multifocal IOLs: By means of an optical system, described by Reiner, images of intraocular lenses can be projected into the eye ("optical implantation"); thus, the optical performance of IOLs can be judged subjectively. Using this method, the refractive 2- and 3-zone models performed best within the multifocal group (contrast sensitivity not significantly worse than that of monofocal IOL), when viewing a low-contrast chart (Regan 4%). All other multifocal lenses (diffractive, aspheric/spherical, refractive 5- and 7 zone models) were significantly inferior to the monofocal IOL. CONCLUSIONS: Implantation of multifocal IOLs should presently be restricted to special indications, particularly to the distinct patient request to dispense with wearing near or bifocal glasses, if possible. Because of the reduction in contrast sensitivity and mesopic vision and the increased glare sensibility, multifocal IOLs should not be implanted especially in professional car drivers. There are, however, differences in optical performance between the various multifocal IOL types. Further improvements, in particular concerning lens technology, will presumably extend the present spectrum of indications. PMID- 8411890 TI - [Chemical and thermal eye burns in the residential area of RWTH Aachen. Analysis of accidents in 1 year using a new automated documentation of findings]. AB - BACKGROUND: Until now there were no statistical data on the incidence and the prevalence of eye burns. Therefore we studied all patients coming to our hospital from the area of Aachen with eye burns during the time from September 1990 until August 1991. PATIENTS AND METHODS: All patients underwent a standardized examination including special history of the burning agent, industrial medical aspects and the employers liability insurance association. The documentation of the anterior eye segment pathology was scored separately for each eye encoded by location and special items. This documentation was worked up by the aid of a database (Filemaker II). 171 patients with eye burns were documented during one year. 65 patients had both eyes burned resulting in 236 documented records. RESULTS: The 171 accidents can be divided in 104 (61%) industrial accidents and 64 (37%) housework accidents. 3 accidents were of unknown origin. Classification of burns was scored according to Reim 1991. 208 (88%) eyes showed score I burns, 27 (11.5%) score II and only one patient showed a score III eye burn. We saw 121 (70%) male patients, 39 (23%) female patients and 11 children (7%). The main age group ranged from 16-45 years. 28% (n = 30) of all accidents happened in machine factories, which have thereby in our study the highest incidence of industrial accidents. CONCLUSION: By means of a one-year statistic we found that over 60% of all eye burns were industrial accidents, 28 in machine factories and 20% in service industries. 37% houseworks accidents are very difficult to prevent because of a deficit of safety rules. PMID- 8411891 TI - [Comparison of complications after intra- and extracapsular cataract extraction with lens implantation. Results of a prospective, randomized, clinical study]. AB - BACKGROUND: The postoperative complications of ICCE with ACL implantation are compared with those of ECCE and PCL. Our clinical experience with ICCE and ACL implantation can not confirm the widespread rejection of this method. PATIENTS AND METHOD: A prospective, randomized, clinical study with participation of medical statisticians was performed. A total of 190 patients with ICCE and ACL and 170 patients with ECCE and PCL were followed up for 2 years. The follow-up examinations were performed upon dismission from the hospital, after 6, 12 and 24 months. The data were compiled in a computer program designed for this study and evaluated by the statisticians. The surgical procedures and the surgeons were defined prior to the beginning of patient recruitment. RESULTS: ICCE with ACL shows much less postoperative complications as usually emphasized. There were only 2 (1.2%) of retinal detachment and no case of corneal decompensation. Cystoid macular edema 8 (4.7%), postoperative vitreous prolaps into the anterior chamber 4 (2.3%) and spontaneous complaints of pain 16 (9.4%) occurred in a low percentage after ICCE with ACL. These complications did not occur after ECCE with PCL. The patients with ECCE and PCL showed capsular fibrosis in 48 (28%) making it the most frequent complication of the whole study. 33% of these patients required YAG-laser capsulotomy. Since retinal detachment occurs in 2.5% after YAG laser capsulotomy we can not regard capsular fibrosis as a totally harmless complication. It is noteworthy that visual acuity is almost identical 1 year after surgery in both methods. CONCLUSIONS: The results of this study show that the evaluation of ICCE with ACL is too negative. The elimination of postoperative complications in this method is more difficult. ECCE with PCL is burdened by frequent capsular fibrosis. Visual acuity is almost the same in both methods 1 years after the operation. ACL-implantation remains our method of choice for secondary implantation in patients with an intact iris diaphragm. PMID- 8411892 TI - [Ultrastructural and enzyme histochemical studies of regional structural differences within the ciliary muscle in various species]. AB - BACKGROUND: The ciliary muscle in man serves two different functions, namely accommodation and regulation of aqueous outflow. It is still not known whether both functions are combined or whether they can also be fulfilled independently from each other. (The latter could provide the possibility of an isolated pharmacological influence on the outflow-related function which should certainly be of use e.g. in glaucoma treatment). MATERIAL AND METHODS: To investigate the presence of functionally different muscle portions within the ciliary muscle and its relation to accommodation we have studied the ciliary muscle of various species showing no (rat, rabbit), moderate (cattle) and good (cat, tupaia glis) accommodative activities. For that purpose enzyme histochemical methods were used which are normally applied for differentiation of skeletal muscle fibers into fast phasic type II-fibres and slow tonic type I-fibres. Additionally, electron microscopical studies were undertaken to evaluate the ultrastructure of the muscle cells. RESULTS: It was found that only those species showing accommodation, were characterized by slight (cattle) or pronounced (cat, tupaia glis) differences in histochemical staining and ultrastructure of muscle cells. Characteristically, the longitudinal portion showing structural affinity to the aqueous outflow system, was different from the inner reticular and circular portions showing more relation to accommodative functions. CONCLUSIONS: These differences might indicate that two different functional systems within the ciliary muscle do exist which have been developed during evolution of higher accommodative mechanisms in the eye. PMID- 8411893 TI - [Mathematical presentation of postoperative regular corneal astigmatism]. AB - BACKGROUND: There are several contradictory approaches to describe a change in corneal astigmatism induced by operations. METHODS: Assuming a rotational ellipsoid for the outer surface of the cornea we derived formulas that represent the change in astigmatism. Also, the change of the corneal power in the axis of the intervention (e.g. operation) has been determined. RESULTS: The exact derivation yield formulas for the absolute value of the astigmatism change identical to those of Naylor and Jaffe but includes, in addition, algorithms for "with-the-rule" and "against-the-rule" components. The spherical change of the cornea is also obtained. The models of Cravy and Naeser are mathematically inconsistent and should be avoided. CONCLUSION: The presentation of the change in astigmatism should depend on the primary question. In many cases, the complicated non-linear vector calculation is not necessary since the actual changes in corneal shape may be demonstrated by simpler formulas. The results of these formulas are much easier to interpret. PMID- 8411894 TI - [NADPH-D reactive choroid ganglion cells in the human]. AB - BACKGROUND: The demonstration of uveal ganglion cells has been investigated so far by silver impregnation, rather complicated neurohistochemical methods and by electron microscopy. We present the first results of a simple method in flat preparation of the human choroid. MATERIAL AND METHODS: Donors' eyes for cornea transplantation, 5 men, 3 women, aged 45 +/- 13 years, with postmortal time of preparation of 12 +/- 9 h (1 h-24 h) were used. After removal of the cornea with scleral ring, flat-preparations of the choroid were treated by the standard Nicotinamide-Adenine-Dinucleotide-Phosphate (NADPH)-diaphorase reaction, mounted on coated slides in Kayser's glycerol-gelatin and observed by light microscope. RESULTS: NADPH-diaphorase reaction reveals abundant aggregates of ganglion cells, with nets of dendrites and axons in the otherwise unstained flat-preparation of the human choroidea. They are more abundant in the layer close to the sclera. CONCLUSION: NADPH-diaphorase reaction is a simple method for the detection of ganglion cells in the human choroid. Their unexpectedly high number suggests yet unknown ganglional functions in the uvea. PMID- 8411895 TI - [Orbital metastasis of a carcinoid tumor of the small intestine]. PMID- 8411896 TI - [Uniform requirements for manuscripts submitted to biomedical journals. International Committee of Medical Journal Editors]. PMID- 8411897 TI - [Urinary tract infections in childhood. Old and new aspects]. AB - Urinary tract infections are one of the most frequent infections during childhood. About 5% of all girls and 0.5% of all boys are suffering from at least one urinary tract infection until the end of schooltime. While the boys predominate in the first year of life with decreasing incidence of infection later on the girls remain predisposed up to an age of twelve. Each pediatrician faces every day the problem of a quick diagnosis and an appropriate therapy of urinary tract infections. While symptomless bacteriuria and cystourethritis do not destroy the renal parenchyma recurrent pyelonephritis may cause irreversible parenchyma scars up to a dialysis demanding renal failure. Besides the well-known mechanical risk factors (i.e. obstruction, reflux) functional risk factors like impaired bladder function alone as well as combined with mechanical ones are gaining in significance. Another factor predisponing to recurrent urinary tract infections is a local immunologic impairment of the urinary tract (i.e. P1-blood group antigen, decreased urothelial function, increased urothelial colonisation). Age-appropriate and individual diagnosis and treatment are necessary first to avoid late damages and second to protect the children against unnecessary diagnostic and therapeutical managements. PMID- 8411898 TI - [Stroke in the child: etiology, differential diagnosis, relevant diagnostic procedures and therapeutic possibilities]. AB - Strokes in children occur in conjunction with arteriovenous malformations or with occlusive vascular diseases secondary to cardiac disease, intracranial infection, hematological and metabolic disorders. Recently several inborn errors of metabolism have been recognized as possible causes of strokelike events and should be considered in differential diagnosis. In a survey we describe our experience with stroke and strokelike episodes in childhood and discuss the approach selected for its management. PMID- 8411899 TI - [Benign intracranial hypertension in childhood--pseudotumor cerebri]. AB - Benign intracranial hypertension-pseudotumor cerebri (PTC) is a rare disease in childhood. We report about our experience in five children with PTC, aged two to ten years. The main symptoms were headache, palsy of VI.th cranial nerve and papilledema. Neuroradiologic studies showed normal ventricles in four and a slit ventricle in one. Enlarged optic nerve sheaths were found in one child prior to therapy. The cerebro spinal (CSF) fluid pressure varied from 380 mm to 480 mm CSF. Four children received acetazolamide, one child dexamethasone. Permanent visual deficit did not occur in any child. PMID- 8411900 TI - [Acute spinal syndromes in children and adolescents]. AB - 23 children with acute spinal syndromes are presented. The clinical signs were caused by trauma in 11 patients, by extramedullary tumors in 6, by operations (coarctation of the aorta, scoliosis) in 4 and by encephalomyelitis in 2. Motor signs (flaccid paraplegia or paraparesis, monoplegia, bilateral weakness) and changes of reflex intensities were always observed. Most children presented with disturbances of sensory functions and micturition, some exhibited ataxia. In addition to the neurological signs, plain films of the spine were of diagnostic importance after trauma, for tumors myelography and/or computerized tomography were most useful. The unfavourable prognosis of posttraumatic paraplegia may be improved by early application of corticosteroids in very high doses. The value of GM1-ganglioside is controversial. In cases of external compression by tumors a decompression of the spinal cord is urgent. "Spinal cord monitoring" can be used intraoperatively in surgery of scoliosis and coarctation of the aorta to detect neurological complications early. Differential diagnosis and therapies of encephalomyelitis and Guillain-Barre-Syndrome are discussed. PMID- 8411901 TI - Constitutional chromosome anomalies in patients with cerebral gigantism (Sotos syndrome). AB - Two boys are presented with the clinical features of cerebral gigantism and chromosomal variants which have not been described so far in this syndrome. In the first boy a de novo pericentric inversion of chromosome Y was found, the karyotypes of all other investigated family members were normal. The patient had an obstructive hypertrophic cardiomyopathy and atrial septal defect type II. The second boy had inherited pericentric inversion of the heterochromatic region of chromosome 9 from his mother. This chromosome 9 variant was also found in his sister who had a similar phenotype but without gigantism. Endocrine evaluation demonstrated normal results in both boys. The intellectual achievement in both cases was average. PMID- 8411902 TI - [Silent persistent ductus arteriosus]. AB - We report on 14 observations undertaken by ourselves of silent persistent ductus arteriosus (SPDA). During a three year period with the routine use of color doppler flow evaluation we found cases of SPDA in 14 children. A very little ductus arteriosus is already known to us through earlier heart catheter examinations which were undertaken because of other isolated heart defects (16 cases in 345 isolated heart defects over 10 years). We also consider the fact that SPDA is not an infrequent result after the occlusion of a patent ductus arteriosus with the Rashkind-Occluder-System and after operative ligatures of patent ductus arteriosus. Finally, we compare our results concerning the frequency of SPDA with current literature and discuss consequential therapies. PMID- 8411903 TI - [Percutaneous sclerotherapy of varicocele--results in children and adolescents]. AB - 65 percutaneous transvenous sclerotherapy of the internal spermatic vein in 58 children and adolescents with varicoceles are reported. Their age ranged from 9.4 to 18 years (mean 14.7 years). The success rate of the sclerotherapy amounted 91.4% while 8.6% recidivations were seen. In three patients slight complications occurred (2 extravasations, 1 spasm of the spermatic vein) which required no further therapy. By repeating the sclerotherapy finally 94.8% of the patients were treated successfully. PMID- 8411904 TI - [Long-term follow-up of infants with pathological gastroesophageal reflux]. AB - Between 1981 and 1984 330 infants with suspected gastroesophageal reflux (GER) were investigated by combined esophageal manometry and pH-monitoring. GER of 283 patients with a manometric reflux time more than 1% was classified as pathologic. After one year of conservative therapy a follow-up-study mainly by 24-hour-pH monitoring could be performed in 108 patients. Two thirds (76 out of 108) still showed a pathological GER. In the age of about two years follow-up-investigation in 41 patients showed in one third of them (13 out of 41) a pathological GER. 5.4% of all investigated children (18 out of 330) hat to be operated because of complicated GER. Follow-up has been performed in 24 children over five years with manometry and/or pH-monitoring and/or barium swallow and/or endoscopy with biopsy. After one year of conservative therapy more than two thirds of them (19 out of 24), after two years one third (eight out of 24) showed a pathological GER. In a few cases (four out of 24) the pathological GER disappeared spontaneously in the age of four to five years. In the age of 1.5 to three years six children had to be operated because of complicated GER. We conclude from this study that a spontaneous healing of pathological GER occurs in about two thirds of the patients older than one year. In about 8% (two out of 24) a pathological GER persists over the fifth year of life representing a permanent risk of GER complications. PMID- 8411905 TI - [Direct genotype analysis in congenital myotonic dystrophy with an unusual family anamnesis]. AB - We report a case of congenital myotonic dystrophy (CMD) in which not only the mother but also the paternal family is affected by myotonic dystrophy (DM). Clinical symptoms consisted of poor spontaneous movements, typical facial appearance, respiratory insufficiency attributable to diaphragmatic weakness, feeding difficulties due to impaired gastrointestinal tract motility and poor sucking, joint contractures and thin ribs. She died at 7 month of age, still ventilated, from aspiration pneumonia. By employing molecular genetic methods we were able to show that the affected child was not homozygous for the DM gene. PMID- 8411907 TI - What decides: high mortality in paediatric acute renal failure? AB - Forty paediatric cases of A.R.F. (Acute Renal Failure) of various aetiology were included in the study. 60% of patients were less than 4 years of age with male predominance. 80% cases reported to us very late with oligoanuria of more than 24 hours (2-7 days). Diarrhoea, vomiting and fever were other dominant symptoms. Maximum cases were severely anaemic (87.5%) with mean Hb 7.73 +/- 1.9 gm%. 40% cases were of underweight while only one case (2.5%) was of over weight, inspite of volume excess in 40% cases. All 24 cases, who were estimated for serum albumin, found to have marked hypoalbuminemia. Mortality was found to be as high as 65% inspite of effective peritoneal dialysis in all cases. High mortality seems to be due to profound anuria of many days (because of marked delay in reaching the hospital), fever and malnutrition besides other factors as aetiology. PMID- 8411906 TI - [Menstrual toxic shock syndrome]. AB - A 15 year old girl presented twice with severe symptoms of multiorgan disease during a period of 4 weeks resulting in intensive care necessary to control illness. According to case definition criteria, a toxic shock syndrome was diagnosed on second admission. This could be confirmed by proof of TSST-1. Beside antibiotics, no specific treatment was used and complete clinical recovery occurred with symptomatic therapy. Discussing the menstrual toxic shock syndrome we point out different aspects of the pathogenesis of the disease including the possible role of tampon use. With regard to the results of several investigators, clinical symptoms of toxic shock syndrome could be the consequence of an uncontrolled immunologic reaction. Looking at this concept, more effective therapy may be available in the near future. PMID- 8411908 TI - Suppression of antibody responses in vitro by bone marrow cells in rats with hereditary liver injury. AB - The primary antibody responses of spleen cell cultures were investigated for LEC rats, a mutant strain of LEA rats, which are known to develop spontaneous hepatic injuries. Compared to LEA rats, LEC rats were significantly low in responding not only to a T-dependent antigen, SRBC, but also to a B-cell stimulant, LPS. The antibody responses of LEC rats having fed a diet deprived of Cu metal (LEC/Cu-) showed values intermediate between LEA and LEC rats, thus suggesting a recovery from an immuno-deficient state as well as liver damage in LEC/Cu- rats. The addition of LEA bone marrow cells prior to culture to LEA or LEC spleen cells, strikingly enhanced the antibody response, whereas LEC bone marrow cells were totally ineffective, or sometimes depressed the response. These results have suggested a newer concept for a possible correlation between liver-dysfunction and bone marrow cell-functions in LEC rats, both encoded by autosomal genes. PMID- 8411909 TI - Effect of pirenzepine on postoperative gastric secretion. AB - Post surgical stress ulcers of the upper digestive tract, once developed, is difficult to control depending on the primary disease and associated complications. As to the cause of postoperative stress ulcers, decreased defensive factors such as gastric mucosal blood flow and increased gastric secretions have been pointed out. Recently, pirenzepine hydrochloride has been shown to be a specific antagonist of muscarinic cholinergic M1 receptors and a suppressant of gastric acid secretion. Therefore we studied its effect on gastric secretions in postoperative patients. Twenty one patients admitted for abdominal surgery, excluding gastric surgery, were selected and randomly assigned to the pirenzepine group (10 cases) or control group (11 cases). Since the serum half life of pirenzepine is 10 hours, 20mg of pirenzepine was administered intravenously immediately after the operation and twice daily (9 a.m. and 9 p.m.) from postoperative day 1 until day 7. Gastric secretions and gastric pH were measured preoperatively and daily until day 7. In the control group, significant increases in the volume of secretion and significant decreases in gastric pH were observed after the operation. In contrast, the pirenzepine group had a significantly decreased amount of gastric secretion and the gastric pH was higher than those in the control group. Thus we conclude that pirenzepine decreases gastric secretion and increases gastric pH in postoperative patients. Pirenzepine can be regarded as an effective agent for the control of postoperative gastric hypersecretion and possibly a good prophylactic for postoperative stress ulcers. PMID- 8411910 TI - [Individualized care and knowledge of cultural differences in the maternity ward. Better understanding of the foreign mother. Interview by Francoise Taillens]. PMID- 8411911 TI - [Anthropological approach. Mental health and exclusion]. PMID- 8411912 TI - [On compassion]. PMID- 8411913 TI - [Care and laboratory: how the two interact--a diploma paper. Laboratory values- important also for nursing]. PMID- 8411914 TI - [Key qualifications: modern educational philosophy--translated into practice]. PMID- 8411915 TI - [Activities of the Swiss Nursing Association's nursing group for operating room service--an interim balance. 66 standards and 687 sub-standards worked out]. PMID- 8411916 TI - [Kinesics--a successful method. Moving three-dimensionally, caring comprehensively]. PMID- 8411917 TI - [Together for quality]. PMID- 8411918 TI - [Building a bridge]. PMID- 8411920 TI - [Immigrant women in the maternity ward. Understanding otherness]. PMID- 8411919 TI - [Balint prize 1993. Relations between nurses and patients]. PMID- 8411921 TI - [At the bedside]. PMID- 8411922 TI - [Results of the questionnaire action on non-nursing activities]. PMID- 8411923 TI - [Nursing aspects in the treatment of patients with adult respiratory distress syndrome]. PMID- 8411924 TI - [Love instead of valium]. PMID- 8411925 TI - [Not enough hands and feet... The clinical emergency]. PMID- 8411926 TI - [Freka Cyst handling study. On the clinical use of the Freka Cyst Set and alternative possibilities]. PMID- 8411928 TI - [Function of the intestine-associated immune system]. PMID- 8411927 TI - [Prevention of transurethral catheter-induced urinary tract infection]. PMID- 8411929 TI - [Medicine, money and ethics; where surgery meets its limits]. PMID- 8411930 TI - [Remaining in flux]. PMID- 8411931 TI - [Prevention of thrombosis in immobilized patients]. PMID- 8411932 TI - [Decubitus ulcer--not necessary!]. PMID- 8411933 TI - [The importance of preventive nursing measures in the prevention of decubitus ulcers]. PMID- 8411934 TI - [Refuse collection systems in the hospital]. PMID- 8411935 TI - [Nosocomial hepatitis B in capillary blood collection for blood sugar determination by reflectometric interpretation]. PMID- 8411937 TI - [Testing the hygienic way of operation of dishwashers in hospitals]. PMID- 8411936 TI - [The clinical emergency. Not enough hands and feet... Part 2]. PMID- 8411938 TI - [Acupuncture nurse]. PMID- 8411939 TI - [Male-female relationships in the nursing professions]. PMID- 8411940 TI - [Munchhausen syndrome in England. Murders in a pediatric hospital]. PMID- 8411942 TI - [The clinical emergency]. PMID- 8411941 TI - [Pictures in the hospital]. PMID- 8411943 TI - [Amyotrophic lateral sclerosis (ALS)--aspects of a poorly understood neurodegeneration]. PMID- 8411944 TI - [Is there a connection between nutrition and arteriosclerosis?]. PMID- 8411945 TI - [Care of AIDS patients in Sweden]. PMID- 8411946 TI - [Attempts at prevention of fetal alcohol syndrome. Model project in Hannover]. PMID- 8411947 TI - [Evening high school for diabetics--a new partly ambulatory alternative with many advantages]. PMID- 8411948 TI - [The responsibility for good and assured care rests with the nurse]. PMID- 8411949 TI - [Temptation]. PMID- 8411950 TI - [Nursing education gives better quality]. PMID- 8411952 TI - [Unlimited respect]. PMID- 8411951 TI - [Building blocks of a bio-psycho-social hypothesis. Are women psychologically more sensitive?]. PMID- 8411953 TI - [Nursing of incontinent patients]. PMID- 8411954 TI - [Can sterile medical disposable materials be reused in the hospital?]. PMID- 8411955 TI - [The value of fat emulsions in parenteral feeding of patients in intensive care]. PMID- 8411956 TI - [What is meant by clinical antisepsis]. PMID- 8411957 TI - [The central role of the cellular immune system in wound healing]. PMID- 8411958 TI - [Practical management of wounds]. PMID- 8411959 TI - [Immunologic tumor therapy]. PMID- 8411960 TI - [Immunotherapy of renal cell carcinoma from an urological viewpoint]. PMID- 8411961 TI - [The Mannheim Uroband: work in the operating room, music on CD. German rock between kidney and prostate]. PMID- 8411962 TI - [Dealing with demented patients]. PMID- 8411963 TI - [The meaning and purpose of patience]. PMID- 8411964 TI - [Church at the bedside]. PMID- 8411965 TI - Introduction to the age-related diagnosis (ARD) index: an age at presentation related index for diagnostic use. AB - Development and, therefore, age is important in paediatrics. Diagnostically useful data from this journal has been related to age and organized to form an age-related diagnostic (ARD) index. The ARD index is designed for non-expert clinical and laboratory workers to use in the early phases of diagnosis and as an addition to existing diagnostic schemes. Entry to the index is from the age at clinical presentation. Each entry is a sequence starting with clinical and laboratory presentations, clinical course, laboratory key investigations and finally diagnosis with volume and page numbers of the original article, the primary source. Within age groups, entries are grouped by diagnoses with the commonest diagnosis first; this has the effect of roughly but not precisely grouping similar clinical and laboratory findings. PMID- 8411966 TI - Infantile neuronal ceroid-lipofuscinosis (INCL): diagnostic criteria. PMID- 8411967 TI - Early juvenile neuronal ceroid-lipofuscinosis or variant Jansky-Bielschowsky disease: diagnostic criteria and nomenclature. PMID- 8411968 TI - Protracted juvenile neuronal ceroid-lipofuscinosis. PMID- 8411969 TI - Adult type of neuronal ceroid-lipofuscinosis. PMID- 8411970 TI - Human forms of neuronal ceroid-lipofuscinosis (Batten disease): consensus on diagnostic criteria, Hamburg 1992. PMID- 8411971 TI - Psychological symptoms and sleep disturbances in neuronal ceroid-lipofuscinoses (NCL). PMID- 8411972 TI - Observations in institutionalized neuronal ceroid-lipofuscinosis patients with special reference to involuntary movements. PMID- 8411973 TI - Seizures, depression and dementia in teenagers with Batten disease. PMID- 8411974 TI - Movement disorders in neuronal ceroid-lipofuscinoses. PMID- 8411975 TI - Pathoarchitectonic pattern of iso- and allocortical lesions in juvenile and adult neuronal ceroid-lipofuscinosis. PMID- 8411976 TI - Immunolocalization studies of subunit c in late-infantile and juvenile Batten disease. PMID- 8411977 TI - Nosological significance of retinopathies in neurodegenerative disorders with emphasis on Batten disease. PMID- 8411978 TI - The significance of animal models for human ceroid-lipofuscinosis. PMID- 8411979 TI - An animal model of the infantile type of neuronal ceroid-lipofuscinosis. PMID- 8411980 TI - Round-table discussion of animal models of ceroid-lipofuscinosis (Batten disease). PMID- 8411981 TI - Ceroid, lipofuscin and the ceroid-lipofuscinoses (Batten disease). PMID- 8411982 TI - Biology of neuronal dysfunction in storage disorders. PMID- 8411983 TI - Pathogenesis of lysosomal storage disorders as illustrated by Gaucher disease. PMID- 8411984 TI - Storage bodies in the ceroid-lipofuscinoses (Batten disease): low-molecular weight components, unusual amino acids and reconstitution of fluorescent bodies from non-fluorescent components. PMID- 8411985 TI - Molecular basis of lysosomal accumulation of subunit c of mitochondrial ATP synthase in neuronal ceroid-lipofuscinosis. PMID- 8411986 TI - Low erythrocyte plasmalogen and plasma docosahexaenoic acid (DHA) in juvenile neuronal ceroid-lipofuscinosis (JNCL). PMID- 8411987 TI - Hereditary ceroid-lipofuscinosis: methylated amino acids in storage body proteins. PMID- 8411988 TI - Juvenile neuronal ceroid-lipofuscinosis: characterization of the dyslipoproteinaemia and demonstration of membrane phospholipid and phospholipid dependent signal transduction abnormalities in cultured skin fibroblasts. PMID- 8411989 TI - Abnormal processing of carboxy-terminal fragment of beta precursor protein (beta PP) in neuronal ceroid-lipofuscinosis (NCL) cases. PMID- 8411990 TI - Glycoconjugate abnormalities in the ceroid-lipofuscinoses. PMID- 8411991 TI - Elevated levels of neutrophil 4-hydroxynonenal in canine neuronal ceroid lipofuscinosis and human immortalized lymphocytes of NCL patients. PMID- 8411992 TI - Reduced phospholipase activity, peptide storage and the pathogenesis of canine neuronal ceroid-lipofuscinosis. PMID- 8411993 TI - Refined assignment of the infantile neuronal ceroid-lipofuscinosis (INCL) locus at 1p32 and the current status of prenatal and carrier diagnostics. PMID- 8411994 TI - Linkage analysis of late-infantile neuronal ceroid-lipofuscinosis (CLN2) using markers on chromosome 16p. PMID- 8411995 TI - Morphological approaches to the prenatal diagnosis of late-infantile and juvenile Batten disease. PMID- 8411996 TI - Prenatal diagnosis of infantile neuronal ceroid-lipofuscinosis, INCL: morphological aspects. PMID- 8411997 TI - Three independent mutations in the same exon of the PCCB gene: differences between Caucasian and Japanese propionic acidaemia. AB - Propionic acidaemia is an inborn error of organic acid metabolism caused by deficiency of propionyl-CoA carboxylase (PCC). Enzyme deficiency can result from mutations in either of the non-identical alpha- and beta-subunits. We have screened genomic DNA from patients with defects in the beta-subunit from two ethnic groups (Caucasians and Japanese) and detected three types of mutations in the same exon of the coding sequence of the beta-subunit: an insertion/deletion that replaces 14 nucleotides with 12 nucleotides of unrelated sequence and eliminates an Msp I site; a 3-bp deletion of a single isoleucine codon immediately proximal to that Msp I site; and a C-->T transition in the same Msp I site. The insertion/deletion was detected only in Caucasian patients in 11 of 34 mutant alleles; the C-->T transition was found only in Japanese patients in 4 of 12 mutant alleles. Following digestion of genomic DNA by Msp I, both of these mutations were detected on Southern blots by the presence of a 2.7-kbp band; they can be distinguished from one another by allele-specific oligonucleotide hybridization following PCR amplification. These results underscore the independent origin of the mutations in the two populations and suggest a key role of this exon in the beta-subunit of PCC. PMID- 8411998 TI - Acylcarnitines in amniotic fluid: application to the prenatal diagnosis of propionic acidaemia. AB - Acylcarnitines were assayed in amniotic fluid by isotope dilution tandem mass spectrometry. Control values for propionylcarnitine, C4 and C5 acylcarnitines were established. Propionylcarnitine was elevated by a factor of 5 relative to controls in the amniotic fluid of three pregnancies affected with propionic acidaemia. This may provide a convenient new method for prenatal diagnosis of propionic acidaemia. One pregnancy at risk for propionic acidaemia but unaffected according to methylcitrate analysis had an intermediate level of propionylcarnitine, indicating the need for further studies to determine the range of concentration in probable heterozygotes. PMID- 8411999 TI - Novel glycine conjugates in medium-chain acyl-CoA dehydrogenase deficiency. AB - The glycine conjugates of isocaproic, 4-methylhexanoic, 7-hydroxyoctanoic and 8 hydroxyoctanoic acids have been identified in the urine of children with medium chain acyl-CoA dehydrogenase (MCAD) deficiency using gas chromatography-mass spectrometry of the trimethylsilyl derivatives. A quantitative study showed that the glycine conjugates of isocaproic and 4-methylhexanoics acids were excreted during acute episodes and in smaller amounts when subjects were asymptomatic. The glycine conjugates of 7-hydroxyoctanoic and 8-hydroxyoctanoic acids were detectable during acute episodes. None of the conjugates was detected in controls or controls receiving a diet containing medium-chain triglycerides. It is suggested that the glycine conjugates of isocaproic acid and 4-methylhexanoic acid are metabolites of branched-chain fatty acids and that they are specific for MCAD deficiency. PMID- 8412000 TI - Cerebral metabolic change after treatment in biotinidase deficiency. AB - A 13.5-year-old boy with biotinidase deficiency was studied 8 days before and 5 months after biotin treatment by positron emission tomography (PET) and computerized electroencephalographic topography (CET). With biotin treatment there was a marked improvement in the presenting symptom of loss of visual acuity and a more modest recovery in spastic quadraparesis. By PET scanning, the relative metabolic rate for glucose was more than 2 standard deviations lower in the temporal and occipital cortices than in adult or age-matched controls. With biotin treatment, these values rose to normal limits for both control groups. By CET, normalized EEG equivalent to the relative glucose metabolic rate showed asymmetric slowing in the left temporal and frontal regions before treatment, whereas none of the 32 leads exceeded normal limits of delta, theta, alpha or beta after treatment. These results suggest a strong correlation between clinical, metabolic and electrical measures of brain function as related to biotin treatment in biotinidase deficiency. PMID- 8412001 TI - Urinary sugar phosphates and related organic acids in fructose-1,6-diphosphatase deficiency. AB - Two sisters with fructose-1,6-diphosphatase deficiency are reported. They presented with ketonuria, elevated plasma transaminase activity and severe metabolic acidosis during hypoglycaemic crises, which resembled Reye syndrome. Intravenous fructose tolerance tests provoked severe hypoglycaemia and metabolic acidosis. Fructose-1,6-diphosphatase activities in both peripheral leukocytes and cultured lymphocytes were below the limit of detection. Urinary organic acid analysis during crises revealed markedly increased excretion of lactate, ketone bodies, glycerol and glycerol-3-phosphate. We newly identified other glycolytic intermediates, glyceraldehyde, 3-phosphoglycerate and fructose-1,6-diphosphate, in the urine during hypoglycaemic attacks or after fructose tolerance tests. Identification of such compounds may be useful in the early diagnosis of this disease. PMID- 8412002 TI - Residual adenylosuccinase activities in fibroblasts of adenylosuccinase-deficient children: parallel deficiency with adenylosuccinate and succinyl-AICAR in profoundly retarded patients and non-parallel deficiency in a mildly retarded girl. AB - Adenylosuccinase (ASase) catalyses both the conversion of succinyl-aminoimidazole carboxamide ribotide (succinyl-AICAR) into AICAR and that of adenylosuccinate into AMP in the synthesis of purine nucleotides. Its deficiency results in the accumulation in body fluids of the nucleosides corresponding to both substrates, succinyl-AICAriboside and succinyladenosine. Two main subtypes of the defect are type I with severe mental retardation and succinyladenosine/succinyl-AICAriboside ratios around 1, and type II with slight mental delay and succinyladenosine/succinyl-AICAriboside ratios around 4. We report that in fibroblasts of type I patients, the activity of ASase with both adenylosuccinate and succinyl-AICAR is about 30% of normal. In contrast, in type II fibroblasts, the activity with adenylosuccinate is only 3% of normal, whereas that with succinyl-AICAR is also 30% of normal. If also present in other tissues, this non parallel deficiency provides an explanation for the higher concentration of succinyladenosine in type II. In type I fibroblasts, ASase is further characterized mainly by a 3-fold to 4-fold increase in Km for succinyl-AICAR, and by retarded elution from an anion exchanger. In type II fibroblasts, ASase is characterized by a similar increase in Km for succinyl-AICAR but by a potent inhibition by KCl and nucleoside triphosphates, and by a normal elution profile. These results suggest a modification of the surface charge of ASase in type I, and the addition of one or more positively charged residues in the active site in type II. PMID- 8412003 TI - Functional studies in fibroblasts of adenylosuccinase-deficient children. AB - In fibroblasts of severely retarded (type I) adenylosuccinase (ASase)-deficient children, activities with the two substrates of the enzyme, succinylaminoimidazole carboxamide ribotide (succinyl-AICAR) and adenylosuccinate are decreased in parallel, to about 30% of normal. In a markedly less retarded (type II) patient, ASase activity with adenylosuccinate reaches only 3% of normal, whereas activity with succinyl-AICAR is also about 30% of normal. To assess the functional significance of a partial versus a profound deficiency of ASase, precursor incorporation studies were performed in intact fibroblasts. In cells from controls and from type I patients, incorporation of 0.2 mmol/L [14C]formate into adenine and guanine nucleotides was not accompanied by accumulation of either [14C]succinyl-AICAR or [14C]adenylosuccinate. Similarly, incorporation of 20 mumol/L [14C]hypoxanthine was not accompanied by accumulation of [14C]adenylosuccinate. In contrast, in fibroblasts of the type II patient, in accordance with the profound deficiency of ASase with adenylosuccinate, and with the inhibitory effect of Cl- and nucleotides on the activity with succinyl-AICAR, incorporation of [14C]formate resulted in accumulation of [14C]succinyl-AICAR and [14C]adenylosuccinate, and incorporation of [14C]hypoxanthine in a marked build up of [14C]adenylosuccinate. That both precursors were still incorporated into the adenine nucleotides of the fibroblasts of the type II patient indicates that adenylate synthesis remains possible even with 3% residual ASase activity, as also shown by their grossly normal ATP concentrations. The results suggest that the pathophysiology of ASase deficiency may be mediated at least in part by accumulation of succinyladenosine and succinyl-AICAriboside. PMID- 8412004 TI - Decreased blood coagulation activities in carbohydrate-deficient glycoprotein syndrome. AB - The carbohydrate-deficient glycoprotein (CDG) syndromes are a newly recognized group of inherited metabolic diseases. We report a Japanese brother and sister with a CDG syndrome. Both patients showed decreased activities of blood coagulation Factor XI and of the coagulation inhibitor protein C. In one of them there was also a somewhat decreased activity of Factor IX and of antithrombin III. Isoelectric focusing of antithrombin III revealed a decrease of negatively charged fractions and an increase of more cathodal bands. Furthermore, there was a discrepancy between activity and antigen level of Factor VIII and protein C. The patients had an incidental deficiency of factor XII. This is the first detailed report on blood coagulation systems in the CDG syndromes. These blood coagulation abnormalities may explain at least in part the thrombotic or haemorrhagic complications of the CDG syndromes. PMID- 8412005 TI - Osteoporosis in lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is an autosomal recessive disease characterized by defective transport of cationic amino acids. Patients have an increased incidence of fractures and their skeletal radiographs show osteoporosis. The aim of the study was to characterize the osteopenia in LPI. Twenty-nine Finnish LPI patients (age range 3.7-44.4 years) were screened for parameters of bone metabolism. Morphometric analysis of bone was carried out in specimens of 9 patients. Collagen synthesis was studied with cultured skin fibroblasts (4 patients) and collagen fibril sizes (3 patients) were measured using electron microscopy. Most histological bone specimens (8/9) showed osteoporosis. Osteomalacia was excluded. Routine clinical laboratory tests were unrevealing. The concentrations of free hydroxyproline and type III procollagen N propeptide in serum and the urinary excretion of hydroxyproline were increased in almost all patients during their growth and in about half of adult patients. Collagen synthesis in LPI fibroblast cultures was significantly decreased compared with that in age-matched controls at 5 (p < 0.01), 14 (p < 0.01) and still at 30 years (p < 0.01), whereas no difference was observed at the age of 44 years (p = N.S.). Osteoporosis in LPI might reflect defective matrix protein synthesis caused by protein deprivation and deficiency of cationic amino acids. Increased collagen turnover can also contribute to the osteoporosis. PMID- 8412006 TI - Improved identification of heterozygotes for phenylketonuria using blood neopterin and biopterin. AB - A novel approach that combines information provided by the metabolism of pteridines and that of phenylalanine has been applied to the detection of heterozygotes for phenylketonuria. Phenylalanine, tyrosine, biopterin and neopterin have been measured in serum from normal controls and heterozygotes for classical phenylketonuria, before and after a phenylalanine oral load. Significant differences in neopterin and biopterin mean values in fasting serum and in the mean increase of biopterin induced by the phenylalanine load were found between groups. Inclusion of pteridine data in the discriminant analysis significantly improved the resolution of the classical phenylalanine loading test for the detection of heterozygotes for phenylketonuria. PMID- 8412007 TI - A fluorimetric enzyme assay for the diagnosis of Sanfilippo disease C (MPS III C). AB - Both the alpha- and beta-anomers of 4-methylumbelliferyl-D-glucosaminide were synthesized and shown to be substrates for the lysosomal acetyl-CoA:glucosaminide N-acetyltransferase. Using the beta-anomer, fibroblasts and leukocytes from 11 different Sanfilippo C patients showed < 1% of mean normal N-acetyltransferase activity. Heterozygotes showed intermediate activities. The enzymatic liberation of the fluorochrome from 4-methylumbelliferyl-beta-D-glucosaminide requires the sequential action of the N-acetyltransferase and beta-hexosaminidase. Normal beta hexosaminidase activity caused complete hydrolysis of the reaction intermediate 4 methylumbelliferyl-beta-D-N-acetylglucosaminide formed by the N acetyltransferase. In cell extracts with a beta-hexosaminidase deficiency, however, a second incubation in the presence of excess beta-hexosaminidase is needed to avoid underestimation of the N-acetyltransferase activity. PMID- 8412008 TI - Biochemical diagnosis of mucopolysaccharidoses: experience of 297 diagnoses in a 15-year period (1977-1991). AB - We report the results over 15 years (1977-1991) for biochemical diagnoses of patients referred from throughout Italy and suspected of having a mucopolysaccharidosis. Of these, 147 patients were diagnosed as being homozygous or hemizygous for a specific lysosomal enzyme deficiency; 74 pregnancies at risk were monitored in their families; 76 heterozygote diagnoses were performed on their relatives, with a total of 48 positive diagnoses. We also report the analysis of genomic DNA from 11 unrelated Italian Hunter patients, using pc2S15 probe. DNA from two patients, digested with Pst-I, showed a variant pattern of hybridization caused by deletion or rearrangement of the gene. PMID- 8412009 TI - Partial HGPRT-deficiency, pheochromocytoma and erythrocytosis. PMID- 8412010 TI - Extracellular and cerebrospinal fluids. AB - The mechanism of formation of extracellular fluid is first described, followed by an explanation of the relation between osmotic force, reflection coefficient and molecular size. The possible mechanism of brain extracellular fluid formation is then proposed in relation to the restriction offered by the blood-brain barrier. The functions and compositions of cerebrospinal fluid (CSF) are then described followed by sections on the process of formation of CSF, the non-electrolytes and proteins in CSF, the drainage mechanisms and protein synthesis by the choroid plexus. PMID- 8412011 TI - The blood-brain barrier: cellular basis. AB - Perfusion experiments with horseradish peroxidase have established that the morphological substrate of the blood-brain barrier is represented by microvascular endothelial cells. They are characterized by complexly arranged tight junctions and a very low rate of transcytotic vesicular transport. They express transport enzymes, carrier systems and brain endothelial cell-specific molecules of unknown function not expressed by any other endothelial cell population. These blood-brain barrier properties are not intrinsic to these cells but are inducible by the surrounding brain tissue. Type I astrocytes injected into the anterior eye chamber of the rat or onto the chick chorioallantoic membrane are able to induce a host-derived angiogenesis and some blood-brain barrier properties in endothelial cells of non-neural origin. Recently we have shown that this cellular interaction is due to the secretion of a soluble astrocyte derived factor(s). Astrocytes are also implicated in the maintenance, functional regulation and the repair of the blood-brain barrier. Complex interactions between other constituents of the microenvironment surrounding the endothelial cells, such as the basement membrane, pericytes, nerve endings, microglial cells and the extracellular fluid, take place and are required for the proper functioning of the blood-brain barrier, which in addition is regionally different as reflected by endothelial cell heterogeneity. PMID- 8412012 TI - Physiology and pathophysiology of organic acids in cerebrospinal fluid. AB - Concentrations of organic acids in cerebrospinal fluid (CSF) appear to be directly dependent upon their rate of production in the brain. There is evidence that the net release of short-chain monocarboxylic acids from the brain is a major route for removing these products of cerebral metabolism. Concentrations of organic acids in blood and CSF are largely independent of each other. Quantitative reference values for the concentrations of organic acids in CSF and plasma as well as ratios of individual organic acids between CSF and plasma were determined in 35 pairs of samples from paediatric patients. Over 25 organic acids were quantifiable in all or in the majority of CSF and/or plasma specimens (limit of detection 1 mumol/L). There were substantial differences in the CSF/plasma ratios between subgroups of organic acids. Metabolites related to fatty-acid oxidation were present in CSF in substantially less amounts than in plasma. Organic acids related to carbohydrate and energy metabolism and to amino acid degradation were present in CSF in the same amounts as or slightly smaller amounts than in plasma. Finally, some organic acids were found in substantially higher amounts in CSF than in plasma, e.g. glycolate, glycerate, 2,4 dihydroxybutyrate, citrate and isocitrate. Studies of organic acids in CSF and plasma samples are presented from patients with 'cerebral' lactic acidosis, disorders of propionate and methylmalonate metabolism, glutaryl-CoA dehydrogenase deficiency and L-2-hydroxy-glutaric aciduria. It became apparent that derangements of organic acids in the CSF may occur independently of the systemic metabolism. Quantitative organic acid analysis in CSF will yield new information on the pathophysiology in the central nervous system (CNS) of these disorders and may prove necessary for successful monitoring of treatment of organoacidopathies, which present mainly with neurological disease. For example, in glutaryl-CoA dehydrogenase deficiency the urinary excretion of glutarate appears to be an inadequate parameter for monitoring the effect of dietary therapy, without plasma and CSF determinations. In L-2-hydroxyglutaric aciduria the elevation of L-2 hydroxyglutarate was found to be greater in CSF than in plasma. In addition, some other organic acids, glycolate, glycerate, 2,4-dihydroxybutyrate, citrate and isocitrate, were also elevated in the CSF of the patients out of proportion to normal levels in plasma and urine. High concentrations of an unknown compound, which was tentatively identified as 2,4-dihydroxyglutarate, were found in the CSF of patients with L-2-hydroxyglutaric aciduria.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412013 TI - Cerebrospinal fluid amino acids, purines and pyrimidines as a tool in the study of metabolic brain diseases. AB - After establishing more extended reference values for amino acids, purines and pyrimidines in cerebrospinal fluid (CSF) in infancy and childhood, we studied 1250 CSF-aliquots from patients who were undergoing a diagnostic lumbar puncture for diverse clinical indications. Our primary aim was to answer the question whether determination of the concentration of amino acids, purines and pyrimidines in CSF is a useful tool in screening for metabolic disorders in children with unexplained mental retardation. In unexplained mental retardation (95 patients) we observed varying abnormalities of CSF. These were reproducible in only 2 patients (a decrease of homocarnosine in combination with two unidentified compounds). Striking abnormalities in pyrimidine content which are limited to CSF are found in argininosuccinic aciduria and uraemia. In uraemia a general decrease in amino acids in CSF and increase of gamma-aminobutyric acid (GABA) was observed. The results obtained indicate that determination of amino acids, purines and pyrimidines in CSF is only of limited value in the diagnosis of unexplained mental retardation. PMID- 8412014 TI - Abnormalities of biogenic amine metabolism. AB - The term biogenic amine is an umbrella term that encompasses all amines with an origin in biological processes. This review will be restricted to the biogenic amine abnormalities that affect the metabolism of serotonin and the catecholamines. The synthesis and catabolism of these neurotransmitters are outlined, and a summary is given of the neurological details, biochemical features, and treatment of the inborn errors that primarily affect their metabolism. An idea is also developed that proposes that abnormalities of biogenic amine metabolism are far more common than is currently considered, and that the search for these problems may be appropriate in any neonate or infant who presents with neurological problems of unknown origin. PMID- 8412015 TI - Non-ketotic hyperglycinaemia: molecular lesion, diagnosis and pathophysiology. AB - Non-ketotic hyperglycinaemia (NKH) is a well-recognized metabolic cause of life threatening illness in the neonate. The fundamental defect is in the glycine cleavage system, which consists of four protein components. Our study revealed that the majority of NKH patients had a specific defect in P-protein (glycine decarboxylase). The primary lesion of NKH at gene level was investigated, using cDNA encoding human glycine decarboxylase. A three-base deletion resulting in deletion of Phe756 was found in a Japanese patient with NKH. The majority of NKH patients in Finland, where there is a high incidence of NKH, were found to be due to a common mutation, a point mutation resulting in the amino acid substitution of Ile564 for Ser564. Prenatal diagnosis is feasible by determining the activity of the glycine cleavage system and is also possible by DNA analysis. Recent findings suggest that a high concentration of glycine in the brain may contribute to the pathophysiology of NKH by overactivating N-methyl-D-aspartate receptors allosterically, which may result in intracellular calcium accumulation, DNA fragmentation and neuronal death. These provide the possibility that early treatment with N-methyl-D-aspartate receptor antagonist may prevent brain damage in NKH. PMID- 8412016 TI - Inherited disorders of GABA metabolism. AB - Gamma-aminobutyric acid (GABA), a major inhibitory neurotransmitter in the mammalian central nervous system, is produced from glutamic acid in a reaction catalysed by glutamic acid decarboxylase. The sequential actions of GABA transaminase (converting GABA to succinic semialdehyde) and succinic semialdehyde dehydrogenase (oxidizing succinic semialdehyde to succinic acid) allow oxidative metabolism of GABA through the tricarboxylic acid cycle. The inherited disorders of GABA metabolism include: (1) pyridoxine-dependent seizures (?glutamic acid decarboxylase deficiency) (> 50 patients); (2) GABA-transaminase deficiency (2 patients/1 family); (3) succinic semialdehyde dehydrogenase deficiency (32 patients/21 families); and (4) homocarnosinosis associated with serum carnosinase deficiency (3 patients/1 family). Homocarnosine is a brain-specific dipeptide of GABA and L-histidine. Of these four defects, definitive enzymatic diagnoses have been made only for GABA-transaminase and succinic semialdehyde dehydrogenase deficiencies. The presumptive mode of inheritance for all disorders is autosomal recessive, and all are associated with central nervous system dysfunction. Only succinic semialdehyde dehydrogenase deficiency manifests organic aciduria, which may account for the higher number of patients identified with this disorder; identification of additional patients with some of the other disorders will require increased request for analysis of cerebrospinal fluid metabolites by paediatricians and neurometabolic specialists. PMID- 8412017 TI - Canavan disease: biochemical and molecular studies. AB - Deficiency of the enzyme aspartoacylase and the accumulation of N-acetylaspartic acid lead to a severe leukodystrophy and spongy degeneration of the brain, Canavan disease (McKusick 271900). Since our discovery in 1988 of the defect in Canavan disease, 144 patients with Canavan disease have been diagnosed in our laboratory. Most of these children are of Ashkenazi Jewish extraction. The level of enzyme activity can be used for carrier testing. Prenatal diagnosis has been difficult using the enzyme assay owing to the low activity of aspartoacylase in cultured chorionic villus samples or amniocytes. The determination of N acetylaspartic acid in the amniotic fluid is another parameter for diagnosis; however, the levels may not always be elevated. Bovine and human aspartoacylase have been purified in our laboratory. Bovine and human cDNA and genomic clones have been isolated and six exons have been localized. This information is being used for the study of Canavan disease at the molecular level. PMID- 8412018 TI - L-2-hydroxyglutaric acidaemia: clinical and biochemical findings in 12 patients and preliminary report on L-2-hydroxyacid dehydrogenase. AB - L-2-Hydroxyglutaric acidaemia represents a newly defined inborn error of metabolism, with increased levels of L-2-hydroxyglutaric acid in urine, plasma and cerebrospinal fluid. The concentration in cerebrospinal fluid is higher than in plasma. The other consistent biochemical finding is an increase of lysine in blood and cerebrospinal fluid, but lysine loading does not increase L-2 hydroxyglutaric acid concentration in plasma. This autosomal recessively inherited disease is expressed as progressive ataxia, mental deficiency with subcortical leukoencephalopathy and cerebellar atrophy on magnetic resonance imaging. Since these features were described in 8 patients by Barth and co workers in 1992, 4 more patients with similar findings have been diagnosed and added to the present series. L-2-Hydroxyglutaric acid is found in only trace amounts on routine gas chromatographic screening in normal persons, and its origin, its fate and even its relevance to normal metabolism are unknown. Therefore its catabolism was studied in normal liver. Incubation of rat liver with L-2-hydroxyglutaric acid did not produce H2O2, which excluded (peroxisomal) L-2-hydroxyacid oxidase as the main route of catabolism. However, L-2 hydroxyglutaric acid is rapidly dehydrogenated if NAD+ is added as a co-factor to the standard reaction medium. This could also be demonstrated in human liver. The preliminary evidence for this enzyme activity in rats and humans, L-2 hydroxyglutaric acid dehydrogenase, is given. Further investigations are required to clarify the possible relevance to the metabolic defect in L-2-hydroxyglutaric acidaemia. PMID- 8412019 TI - Biochemical pathogenesis of subacute combined degeneration of the spinal cord and brain. AB - In humans, subacute combined degeneration of the spinal cord and brain, a primary demyelinating disease, is caused by cobalamin or methyltetrahydrofolate deficiency. Experimental studies into its pathogenesis suggest that dysfunction of the methyl-transfer pathway may be the cause. Compelling evidence for this comes from the study of inborn errors of cobalamin metabolism where deficiency of methylcobalamin, but not deoxyadenosylcobalamin, is associated with demyelination. Recent studies have focused upon inborn errors of the methyl transfer pathway. Cerebrospinal fluid concentrations of metabolites of the methyl transfer pathway have been measured in humans with sequential errors of the pathway and correlated with demyelination demonstrated on magnetic resonance imaging of the brain. This has provided new data suggesting that deficiency of S adenosylmethionine is critical to the development of demyelination in cobalamin deficiency. PMID- 8412020 TI - Inborn errors and demyelination: MRI and the diagnosis of white matter disease. AB - The progress and extent of myelination can be assessed using magnetic resonance imaging (MRI). Myelination is delayed or diminished in several inherited metabolic abnormalities presenting in early life. Only minimal myelination of the CNS occurs in Pelizaeus-Merzbacher disease. Dysmyelination tends to produce fairly symmetrical lesions affecting white matter. In many mitochondrial enzyme and some lysosomal defects, the grey matter is also involved. The appearances and in particular the distribution on MRI and/or CT are characteristic in some conditions and the diagnosis is limited in others. Demyelination due to inflammatory disorders typically causes multifocal white matter lesions, recurrent in multiple sclerosis, monophasic in acute disseminated encephalomyelitis, extending in progressive multifocal leukoencephalopathy and classically involving the pons or corpus callosum in myelinolysis. Hypoxic ischaemic lesions may be metabolically induced and simulate primary demyelinating disorders. Mitochondrial enzyme defects in particular may present with stroke like appearances. In many of these conditions, diagnosis is biochemical, but imaging has a significant role in suggesting the diagnosis, and documenting progression, response to therapy or complications. PMID- 8412021 TI - Genetic analysis of Batten disease. AB - Batten disease, or neuronal ceroid-lipofuscinosis (CLN) comprises a group of inherited neurodegenerative disorders characterized by the accumulation of autofluorescent lipopigment in neurones. The three main childhood varieties- infantile (CLN1), late-infantile (CLN2) and juvenile (CLN3)--manifest autosomal recessive inheritance. The basic biochemical defect remains unknown. The strategy of positional cloning is being pursued to elucidate the molecular basis of Batten disease. The infantile disease locus (CLN1) has been mapped by linkage analysis to human chromosome 1p32, and the juvenile disease locus (CLN3) to human chromosome 16p12. In each case marker loci in strong linkage disequilibrium with the disease loci have been identified. Locus heterogeneity between classical late infantile CLN (CLN2) and both CLN1 and CLN3 has been demonstrated. Work is in progress to clone CLN1 and CLN3 and to map CLN2. Identification of linked markers has provided a new approach to prenatal diagnosis. The methodology exists for positional cloning of these genes and elucidation of the molecular genetic basis of the ceroid lipofuscinoses. PMID- 8412022 TI - Recent developments in Menkes disease. AB - Recent studies on Menkes disease are reviewed, focusing especially on copper transport in the cells. A large amount of copper accumulated in the organelle free cytoplasm, whereas mitochondria were in a state of copper deficiency, indicating that Menkes mutation probably affects copper transport from the cytosol to the organelles in the cells. Microscopic observation of the brain of the macular mouse showed that copper accumulates in the blood vessels. Observation of the brain tissue of the macular mouse after intraventricular administration of copper revealed that copper accumulates in the glia as well as the blood vessels. Copper accumulation was also observed in cultured astrocytes, a type of glial cell, indicating that the affected astrocytes accumulate blood borne copper and release little of it in the patients with Menkes disease. Thus the effective treatment of Menkes disease could possibly be to release trapped copper from the blood vessels and glia into the neurons. PMID- 8412023 TI - Proton magnetic resonance spectroscopy studies in lactic acidosis and mitochondrial disorders. AB - Congenital lactic acidosis form a large group of disorders that are commonly associated with profound neurological dysfunction. Difficulties are frequently encountered in establishing a diagnosis, and the mechanisms underlying brain damage are poorly understood. We have performed proton magnetic resonance spectroscopy (1H-MRS) on 24 patients under investigation for suspected metabolic disorder, and have compared the MRS observations of brain lactate with measurements of cerebrospinal fluid (CSF) lactate. We have shown good concordance between the two types of observation, confirming the value of the CSF measurements. Regional variations in brain lactate are detected in some cases, and these may help to elucidate the mechanisms underlying selective brain damage. PMID- 8412024 TI - Opportunities for enhanced cervical and breast screening referral in Charleston County. AB - In order to increase awareness of the need for pap smear and mammographic cancer screening, the Professional Education Committee of the Charleston County Chapter of the American Cancer Society (ACS) separately surveyed Charleston County women and physicians regarding their use of women's cancer screening tests. Seven hundred and thirty-one women responded to a Cancer Screening Survey for Women. Women not complying with ACS guidelines for pap smears and mammography, when compared to women complying with the guidelines, were found to be twice as likely to be non-white, much more frequently lacked a regular physician or a gynecologist, more commonly lacked insurance paying at least in part for the test, and more frequently reported that physicians visited did not suggest pap smear or mammographic screening. Almost one-third of 75 responding physicians acknowledged not discussing pap smear and mammographic screening with their patients, a practice most often attributed to "specialization." Among physicians not referring women for needed mammograms or pap smears, roughly half also cited patient refusal, deferral, and oversight as reasons. The survey data suggest that a simple, practical, method is needed in doctors' offices to remind women of the need for periodic pap smear and mammographic screening, especially whenever women lacking a regular gynecologist seek any type of medical care. The authors recognize that these samples of females and physicians are not necessarily representative of those components of the Charleston area overall.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412025 TI - Diagnostic office arthroscopy. PMID- 8412026 TI - Combined modality therapy in the treatment of epidermoid carcinoma of the anus at the Medical University of South Carolina. AB - Treatment of anal epidermoid carcinoma by radiation and chemotherapy does offer an excellent opportunity to maintain quality of life. No patient in this series suffered long-term complications; and all have maintained rectal, anal, and sexual function. Patients with early stage lesions (AJC I-II) are most amenable to anus conserving therapy for epidermoid cancer of the anus. The success rate is markedly diminished in patients having more advanced disease, with patients possessing AJC stage III and VI disease presenting major problems. The possibility of post operative radiation therapy is markedly compromised by preoperative radiation therapy. Clinical surveillance and early recognition of these lesions by primary physicians can now offer patients with lesser stage cancer of this type excellent opportunity for organ preservation therapy. For those patients presenting with Stage III and VI carcinoma of the anus the aforementioned problems should be addressed and discussed before initiating conservative therapy. PMID- 8412027 TI - Neural tube defect prevention. The folic acid story. PMID- 8412028 TI - Physical activity and associated risk factors in South Carolina 1988-1990. PMID- 8412029 TI - Physical activity behavior in South Carolina youth. PMID- 8412030 TI - Youth health--the key to lifelong fitness and the challenge to health care in the next century. PMID- 8412031 TI - Physical fitness for adults--a commentary. PMID- 8412032 TI - The need for physical activity in older adults. PMID- 8412033 TI - Exercise as a clinical tool. PMID- 8412034 TI - Physical activity in South Carolina. PMID- 8412035 TI - Content and teaching strategies in 10 selected drug abuse prevention curricula. AB - This paper reports on content and teaching strategies in 10 drug abuse prevention curricula available to public and private schools. While similarities existed among the curricula, they employed a range of content as well as strategies. Interactive teaching strategies needed to implement the curricula include full class discussion, small group activities, brainstorm, and role play. Due to the complexity of the strategies, the curricula may not provide sufficient background information or training for teacher implementation in the classroom. PMID- 8412036 TI - Creating a safe environment for children in daycare. AB - Injuries represent the leading cause of death for children older than age one. Likewise, some 22 million children are injured each year, with more than 1 million requiring medical treatment. More than 40% of children currently receive full-time or part-time day care, but little research has examined the status of injuries to children in daycare. This paper provides information on the nature and extent of injuries to children, and contributing factors responsible for those injuries. One case study of research on injuries in Minneapolis, Minn., daycare centers is discussed. Recommendations for action are offered. PMID- 8412037 TI - Implementing a school-based STD/HIV prevention intervention: collaboration between a university medical center and an urban school district. AB - To effectively implement a school-based STD/HIV prevention intervention program developed by a community-based agency, several issues must be addressed to build a collaborative partnership between the agency and the school district. Steps taken by the Division of Adolescent Medicine, University of California, San Francisco, in achieving partnership with a local urban school district are outlined. The process of recruiting school district administrators, health education liaisons, teachers, parents, and students is emphasized. PMID- 8412038 TI - A model for training community-based providers for children with special health care needs. AB - Children with special health care needs pose many challenges for community-based providers. Programs such as school, child care, and Head Start provide services to children that previously would have been delivered in hospitals or at home. The need for a system of care to assure safe, quality services for these children is paramount. While health procedure manuals for schools have been developed, few recommendations address the actual training of providers or the trainers of providers. This article describes a system of training which includes steps in managing the care of children with special health care needs, levels of training, and expected competencies for the care providers of children with special health care needs. PMID- 8412039 TI - The politics of school-based clinics: a community-level analysis. AB - School-based primary health care, supported as an effective means for addressing the health needs of youth, also tends to be perceived as controversial partly due to opposition by national and state level organizations. This investigation used case studies of four selected school-based clinic (SBC) sites, and of one community in which an SBC was disputed, to investigate the existence of organized opposition, how proponents contended with opposition, and effects of organized opposition on SBC approval and implementation. Four sites experienced opposition affiliated with larger conservative organizations. Opponents' strategies and proponents counterstrategies were identified. Effects ranged from limiting SBC services to preventing an SBC's establishment. Proponents, consisting largely of medical and school administrative professionals, overcame opposition through strategies to increase public awareness about youth health issues and to demonstrate public support in the face of expressed opposition. Implications for planning and implementing potentially controversial programs are offered. PMID- 8412040 TI - Kids, health, and the media: what can public health offer? PMID- 8412041 TI - The role of pet dogs in casual conversations of elderly adults. AB - Casual conversations were recorded as elderly persons routinely walked their dogs through a familiar mobile home park in the United States. Control observations included walks without dogs by owners and non-owners of dogs. All owners talked to and about their dogs. Transcribed conversations indicated that dogs were a primary focus of conversation. A majority of sentences to dogs were imperatives; the owners were instructing the dogs. Dog owners frequently included dogs' names or nicknames in their sentences when they spoke to the dogs and made reference to the dogs' wishes or needs. Speaking to dogs was also associated with frequent repetition of sentences. Passersby talked to the owners about their dogs whether or not the dogs were present. When dog owners spoke with other people, their conversations often concerned activities that were occurring in the present, whereas conversations of non-owners focused on stories about past events. Dog owners reported taking twice as many daily walks as non-owners. Dog owners also reported significantly less dissatisfaction with their social, physical and emotional states. PMID- 8412042 TI - Gender differences in the source and level of self-esteem of Chinese college students. AB - This research examined possible gender differences in the source and level of self-esteem of 99 male and 90 female undergraduates from mainland and China. There was little evidence of a gender difference in the level of overall self esteem, but gender differences were evident in the subjects' ratings of the importance to their self-concept and their self-satisfaction with lower order facets of the self, indicating the necessity for using multidimensional measures of the self and for preserving the self-concept/self-esteem distinction. PMID- 8412043 TI - Beliefs about the world and recovery from myocardial infarction. AB - Seventy male, Hindu Myocardial Infarction (MI) patients were interviewed twice--4 to 5 days after their first heart attack (Time 1) and a month after their first heart attack (Time 2). The patients' beliefs about the world (world beliefs), about the causes of the disease (causal beliefs), and about the factors contributing to their recovery (recovery beliefs) were measured. Each category of beliefs was concerned with three domains: karma, God, and just world (or self). The patients' physical and psychological recovery was evaluated. World and recovery beliefs were intercorrelated, but these beliefs were only weakly correlated with causal beliefs. Furthermore, world beliefs and recovery beliefs were positively associated with recovery from MI at both Time 1 and Time 2. Attribution of causality to God was negatively correlated with medical recovery, perceived recovery, and mood state at Time 2. This trend was in the reverse direction for attribution of causality to self. PMID- 8412044 TI - Juvenile delinquency in the northern states of Nigeria. PMID- 8412045 TI - Dimensions of romantic love in British female adolescents. PMID- 8412046 TI - Involvement of membrane excitation failure in fatigue induced by intermittent submaximal voluntary contraction of the first dorsal interosseous muscle. AB - Factors involving muscle fatigue were studied at intermittent isometric contraction of the first dorsal interosseous muscle (FDI). Subjects made repeated contraction of 6 s, 40% of maximal voluntary contraction (40% MVC) (= target force) followed by 4 s rest until the target force could no longer be maintained. Measurement of MVC, recording of M wave evoked from the relaxed muscle, force measurement during tetanic electrical stimulation and measurement of serum K+ concentration were performed every 2 min during the fatiguing exercise and after 10 min of recovery. The declining rate of MVC force was always smaller than that of the force generated by 50 Hz stimulation, suggesting that no central nervous system (CNS) failure was involved. The amplitude and the area of the M wave declined while the duration of the wave increased, accompanied by the rise in serum K+ concentration. After 10 min of recovery, all parameters except the MVC force were restored. Thus we concluded that the failure in muscle membrane excitation, as well as the failure in excitation/contraction coupling, is involved in the deteriorating force generating capacity of FDI during intermittent isometric contraction. PMID- 8412047 TI - The effect of various recovery modalities on subsequent performance, in consecutive supramaximal exercise. AB - Different recovery strategies from maximal exercise seem to induce different lactate utilization patterns without significantly affecting performance on one subsequent maximal exercise. It remains unclear however, how varying recovery modalities affects repeated maximal exercise. To study this, we examined in 16 subjects, the influence of passive (P), active leg (L) and active arm (A) twenty minutes recovery periods separating a series of four exhaustive exercises, up to two minutes duration. Significant decreases in performance between the first and fourth exercise were observed in all recovery series but a significant decrease in performance in the second exercise was observed during passive recovery alone (p < 0.01). When the different types of recovery are compared, a more pronounced decrement in performance was found during passive recovery when first and last exercises are compared (p < 0.04). Pedaling duration in each successive exercise was unaffected in A or L but was significantly shorter in P (p < 0.03). Highly significant differences in mean blood lactate kinetics were found for the three recovery patterns used, with more elevated peak and nadir levels in passive recovery, intermediate values in active arm and lowest concentrations in active leg recovery. However, no correlation was found between performance and lactate concentration at the onset of exercise (r = -0.15; p = NS). Mean heart rates were similar throughout the experimental protocol except for a lower cardiac frequency during the last 5 minutes of passive recovery (p < 0.01). Blood hematocrits showed higher hemoconcentrations in repeated exercise during passive recovery (p < 0.01) despite significantly lower total fluid losses in this group. A significant correlation between peak hematocrit and blood lactate was also found (r = 0.67; p < 0.001). We conclude that the type of recovery has a significant effect on blood lactate elimination kinetics, and active recovery is beneficial in the preservation of performance during repeated maximal exercise. Furthermore, plasma shifts across the extra and intravascular spaces are induced by maximal exercise, and appear to closely follow blood lactate kinetics. PMID- 8412048 TI - Aerobic characteristics, oxygen debt and blood lactate in speed endurance athletes during training. AB - Aerobic characteristics, oxygen debt and blood lactate were analysed in 20 male speed endurance athletes (400 m sprinters and 400 m hurdlers). The subjects were tested three times; at the beginning of March, at the end of May and at the end of August. Aerobic and anaerobic threshold and maximal oxygen uptake measured on the treadmill decreased (p < 0.05-0.01) from the second test occasion to the third one. The anaerobic work test on the treadmill was a constant load test at 5.56 m.s-1 with a slope of 4 degrees. The time to exhaustion increased (p < 0.05) from the first test occasion (112 +/- 17 s) to the second one (136 +/- 35 s) and did not change in the last test (135 +/- 25 s). Following the anaerobic work test oxygen debt was measured during 20 minutes. The highest total oxygen debt values (144 +/- 19 ml.kg-1) were observed in the second test occasion. Peak blood lactate following the anaerobic work increased (p < 0.05) from the first test occasion to the second one and remained at the same level during the next three competitive months. The good speed endurance athletes differed from the poor counterparts in time to exhaustion (p < 0.01), in 100 m record time (p < 0.01) and in maximal oxygen uptake (p < 0.05). Our results suggest that aerobic characteristics decrease during the competitive period in speed endurance athletes. The anaerobic performance capacity including work time and peak blood lactate is at high level in the competitive period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412049 TI - Nedocromil sodium in the prevention of exercise-induced bronchospasm in athletes with asthma. AB - The aim of this study was to determine the efficacy of nedocromil sodium in the prevention of exercise-induced bronchospasm (EIB) in 13 top athletes affected by bronchial asthma. At a dose of 4 mg the drug significantly reduced the fall in FEV1 compared with placebo but not with respect to basal values. In 9 athletes, 4 mg nedocromil sodium produced a good protective effect and reduced the mean fall in FEV1 to 4% with respect to baseline values, while in the remaining 4 subjects, the protective effect was not satisfactory. In these 4 "non responders" 6 mg nedocromil was effective, and in 2 cases induced prolonged bronchodilatation. In conclusion, the effect of nedocromil sodium in the prevention of EIB may be dose dependent in relation to the degree of bronchial hyperreactivity or to interference of other factors. PMID- 8412050 TI - Effects of alpha 1-receptor blockade on the cardiovascular and thermoregulatory responses to severe exercise in the heat. AB - Up to an internal temperature of 38 degrees C, nonacral skin blood flow increases with mild exercise in a warm environment. While this appears to be due predominately to an active vasodilator system and not to altered vasoconstrictor activity, it is uncertain if more intense exercise affects this vasodilator response. Consequently, six fit (VO2pk = 62.8 +/- 2.2 ml.kg-1.min-1, mean +/- SEM) college aged men performed 30 min bouts of cycle ergometry exercise at 70% VO2pk in a hot environment (35 degrees C, 55% rh) while under the influence of either an alpha 1-receptor blocker (prazosin, PRAZ) or a placebo (CTL). Resting rectal temperature (Tre) was lower (p = 0.002) and heart rate (HR) higher (p = 0.018) during PRAZ. Neither resting nor exercise forearm blood flow (FBF, plethysmography) and forearm vascular conductance (FVC) differed between PRAZ and CTL. Exercise Tre and mean skin temperature (Tsk) did not differ significantly between treatments, rising to 38.7 and 36.4 degrees C, respectively, by the 30th min of exercise. Exercise HR was 4 beats.min-1 higher (p = 0.016) with PRAZ. Mean arterial pressure (MAP) did not differ significantly between drug treatments, As FBF, FVC, Tre, Tsk and MAP were similar between treatments while only HR was significantly affected by PRAZ, it appears that alpha 1-receptor blockade does not alter the thermoregulatory responses to severe exercise in the heat. However, the elevated HR in the presence of a stable MAP suggests that central venous return may have been attenuated during PRAZ. PMID- 8412051 TI - Anthropometric correlates with strength performance among resistance trained athletes. AB - The purpose of this study was to determine the relationship between selected anthropometric dimensions and strength performance in resistance trained athletes. Fifty-eight college football players were measured following the completion of a 10-week resistance training program for one-repetition maximum (1 RM) lifts in the bench press, squat, and dead lift and for 11 anthropometric dimensions. Results indicated that the highest relationships existed between estimates of regional muscle mass (arm circumference, arm muscle cross-sectional area, and thigh circumference) and lifting performance. Multiple regression analysis selected arm size and %fat as variables common to the prediction of all three lifts. The fewer joints and muscle groups involved in a lift, the greater the predictive accuracy from structural dimensions. It was concluded that body structure and conformation make significant contributions to maximum strength performance in highly trained strength athletes. PMID- 8412052 TI - Relationships among swimming performance, body composition and somatotype in competitive collegiate swimmers. AB - Relationships were determined between sprint swimming performance and estimates of body composition and somatotype components in competitive collegiate swimmers. Forty-three women and 31 men underwent anthropometric and hydrodensitometric measurements at the beginning of a competitive season. There were significant partial correlations (swim stroke was partialed out) between swimming performance (time in a competitive 100-yard swim of each swimmer's major competitive stroke) and height (-0.466, p < 0.01), the mesomorphic (0.404, p < 0.01) and ectomorphic (-0.398, p < 0.01) components of somatotype, percent body fat (0.351, p < 0.05), and fat-free weight (-0.332, p < 0.05) among the women, but no significant correlations among the men. Twenty-three of the women and 21 of the men were also measured and timed at the end of the competitive season. Again, there were significant (p < 0.01) partial correlations (season and stroke were partialed out) between swimming performance and height (-0.766), fat-free weight (-0.657), body weight (-0.437), and the ectomorphic (-0.441) and mesomorphic (0.392, p < 0.01) components of somatotype in women, but no significant correlations among the men. These findings indicate that measurements of body composition and somatotype may be predictors of swimming performance in women but not in men. PMID- 8412053 TI - Differences in estimates of percent body fat using bioelectrical impedance. AB - The purpose of the present investigation was to analyze the replicability of percent body fat (% fat) determined from Bioelectrical Impedance Analyses (BIA) after 2 different postprandial time periods and by 2 different technicians. The BIA % fat measures were also compared to those determined from skinfold measurements (SF) and hydrostatic weighing (HW). Fifteen physically active men had body composition determined on 4 separate days over a 2 week period under controlled conditions, once each week after fasting for 12 hours and once after 3 hours. Each day the subjects had BIA and SF measures taken by different trained technicians. Once each week the subjects also were hydrostatically weighed (HW). There were no differences for either BIA or SF between the 2 postprandial time periods. There were significant differences between technicians for both BIA and SF (p < 0.01), although the relationships between technicians were good (r = 0.90 and 0.95, respectively). Mean values for BIA (11.8 +/- 3.2%) and HW (11.0 +/- 7.2%) were similar, however, the correlation was only r = 0.50. These results indicate that although BIA gave relatively consistent values, it regressed both low and high % fat values towards the mean of the sample. PMID- 8412054 TI - The morphotype in a group of peri-pubertal soccer players. AB - Few morphological data are available for adolescent soccer players (SP), a lack that could affect young-age talent selection. In order to collect lacking data for SP, 50 performers aged 13.1 +/- 0.5 years old were checked in one of the main "vivariums" of talented "A League" SP. All subjects were measured according to the Heath/Carter anthropometric somatotype method and subsequently, the results were compared with those found in relevant literature for adolescents. To gain further insight, the sample was subdivided into two classes, according to their performance level: beginners (B, n = 26) and those experienced (E, n = 24). Between B and E subjects significant differences were found for: weight, abdominal skinfold, calf circumference, body density (0.05 < p < 0.01); height, tricipital skinfold, humerus biepicondylar width, ponderal index, LBW, endo and ectomorph components (p < 0.01). B somatotype resulted to be: 1.6-4.3-3.5; while E: 2.2-4.5-2.9. Both in E and B sub-groups the distribution of somatotypes according to their component dominance revealed to be rather similar, indicating a substantial homogeneity inside the subgroups. When compared with adult SP, the whole sample shows a morphotype that is already adapted for soccer. Observing this, it is possible to infer that our adolescent group contains the morphotype needed to perform well (and probably to excel), in the considered sporting activity. PMID- 8412055 TI - Accuracy of physical working capacity (PWC170) in estimating aerobic fitness in children. AB - Physical working capacity, the workload at a heart rate of 170 bpm (PWC170), has been utilized as a marker of maximal oxygen uptake. The precision of PWC170 in predicting VO2max in children has not been fully evaluated. In this study, 35 children (18 boys and 17 girls, mean ages 10.5 and 9.9 years, respectively) underwent maximal cycle testing to assess the relationship between VO2max and PWC170. These measures correlated closely in absolute terms (r = 0.71 and 0.70 for girls and boys, respectively), but the relationship was weaker when both were expressed per kg body weight (r = 0.65 and 0.48, respectively). When VO2max was calculated from the regression equation of VO2max versus PWC170, the mean error from measured VO2max was 3.4 ml.kg-1.min-1 (SD 2.5) for the girls and 2.8 ml.kg 1.min-1 (SD 2.6) for the boys. These findings indicate that although mean predictability of VO2max from PWC170 is good, the variability is wide, with a 10 15% error at one standard deviation. PWC170 provides only a crude estimate of VO2max and should not be used to predict individual maximal aerobic power. PMID- 8412056 TI - Exercise dependence in relation to competitive orientation of runners. AB - An increasing number of runners seem to be vulnerable to the exercise dependence (ED) syndrome, a clinical disorder in which the athlete is unwilling to alter or discontinue training regimens despite medical contraindications to continued training compliance. As habituation to a stimulus has been regarded to be a diagnostic criterion of an addicted state, we hypothesized that performance in running events of increasing distances would be accompanied by an increased tendency towards ED. To examine this relationship, an exercise addiction scale was completed by recreational (non-competitive) runners (N = 33), five kilometer (5K) runners (N = 24), marathoners (N = 32), and ultramarathon (50 miles) runners (N = 61). ANOVA procedures indicated that both marathoners and ultramarathoners showed significantly higher ED scores [3.78 +/- 1.6 (SD) and 4.59 +/- 1.8, respectively] as compared to 5K (2.9 +/- 1.0) and recreational runners (2.16 +/- 1.3) (p < 0.05). In addition, ultramarathoners presented significantly higher ED scores than those of marathoners (p < 0.05). The data indicate that a tendency towards ED may motivate participation in competition of increasing distance and support a habituation effect consistent with acquired dependence behavior. PMID- 8412057 TI - Relationship between hamstring strains and leg muscle strength. A follow-up study of collegiate track and field athletes. AB - The purpose of the present study was to assess the relationship between hamstring strains and leg muscle strength. The bilateral isometric extensions and flexion maximum voluntary contraction (MVC) of knee and hip were measured among 64 collegiate track and field athletes (128 legs). The values of MVC per body weight, flexion to extension ratio and bilateral legs imbalance index were calculated as the parameters for the investigation. The follow-up research was performed within the following two years. Among the 64 subjects (128 legs): 26 subjects (31 legs), 24.2 percent had suffered from hamstring strains. Then, the subjects were divided into injured (31 legs) and uninjured (97 legs) groups respectively. The parameters of the lower extremities measured at the beginning were compared for the two groups. The different rates of the hip flexion and knee extension of bilateral legs of the injured group were significantly higher than those of the uninjured group (p < 0.05). In the injured group, the value of MVC per body-weight of the knee flexor and the flexion-extension ratio were significantly lower than in the uninjured group (p < 0.05). In conclusion, the imbalance of the bilateral legs, the hamstring strength and the ratio of the flexor to extensor were shown to be parameters related to the occurrence of hamstring strains. PMID- 8412058 TI - High density lipoprotein cholesterol following anaerobic swimming in trained swimmers. AB - HDL-C, LDL-C, total cholesterol and glycerol were determined in venous blood of 10 male trained swimmers at rest and following 100-m anaerobic swimming. When compared to rest levels, the peak HDL-C and glycerol concentrations were significantly enhanced following anaerobic swim test (p < 0.01). However, peak LDL-C and total cholesterol after anaerobic swim were not significantly increased compared with the rest level. The present data demonstrated that the anaerobic swimming induces an increase in HDL-C metabolism, suggesting that the anaerobic exercise per se was one reason for the elevated HDL-C levels. PMID- 8412059 TI - Hemodynamic responses of trained and sedentary pregnant women to semi-supine cycling. PMID- 8412060 TI - Effect of endurance physical training on cardio-respiratory system reactive features (mechanisms of training load cumulation influence) AB - The results of a series of experiments carried out under heavy physical loads during the training of 96 elite cyclists, rowers and swimmers were based on the following: (1) complex analysis of dynamics of sensitivity, stability and speed of responses of cardio-respiratory system (CRS) to hypoxia, CO2-H+ ionic irritants; (2) neurogenic (working) response stimuli; (3) range of changes in ventilatory, circulatory and metabolic responses and their dynamic characteristics. It was demonstrated that sensitivity, stability and speed of responses to shifts in respiratory homeostasis changed during each physical load and in the process of training. The nature and the extent of changes are connected with the intensity of the load as well as its type and duration. After training, we observed changes in the sensitivity, stability and response speed of CRS to shifts in respiratory homeostasis; we also noted that the total reactivity changes revealed the same direction of change as the above mentioned CRS physiological reactivity parameters. It was also observed that the dynamics of reactivity changes to hypoxia, CO2-H+ irritants and neurogenic CRS response are connected to the dynamics of peak of CRS response to physical load and to its stability and response speed. These results indicate the existence of certain mechanisms which regulate the setting up of the functional capacity in athletes. PMID- 8412061 TI - Protective effect of 2-chloroadenosine on lung ischemia reperfusion injury. AB - Reperfusion following ischemia yields an inflammatory response characterized by polymorphonuclear leukocyte (PMN) influx, inflammatory mediator release, microvascular permeability alteration, and protein-enriched fluid transudation. Evidence has accumulated suggesting that low-dose adenosine may "down-regulate" the PMN response. This study evaluated the effects of an adenosine analogue, 2 chloroadenosine (2CA), on ischemia-reperfusion (IR) injury in rabbit lungs. In these experiments the left pulmonary hilum was skeletonized, obliterating the bronchial circulation, and the left pulmonary artery and vein were occluded for 1 min for the sham ischemia (SI-V) group or for 1 hr for the ischemia (I-V) and 2CA treated (I-A) groups. The left lung was inflated with nitrogen during the ischemic period. Saline (SI-V and I-V groups) or 2CA (I-A group) infusions were begun prior to and during the reperfusion period. After 4 hr of reperfusion and restored ventilation, selective left lung physiologic measurements and bronchoalveolar lavage (BAL) were performed. Groups (N = 8/group) were compared using analysis of variance. The I-A group demonstrated a significantly lower mean pulmonary artery pressure and higher cardiac output than the I-V group. Pulmonary vascular resistance was significantly elevated in group I-V compared to group I A. A significantly greater alveolar WBC influx and protein transudation (BAL/plasma albumin) occurred in the ischemic group compared to the 2CA-treated animals and sham controls. Decreased PaO2 and increased venous admixture were noted in the ischemic group, but did not reach significance when compared to the 2CA group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412062 TI - Reduction of experimental vein graft intimal hyperplasia by ketanserin. AB - Intimal hyperplasia is considered to be the result of smooth muscle cell proliferation. Experimental vein grafts (VG) in the rabbit develop intimal hyperplasia and a contractile response to serotonin (5-HT), mediated by a 5-HT2 receptor. 5-HT is known to stimulate smooth muscle cell proliferation in vitro and therefore may be linked to the development of intimal hyperplasia. This study examines the effect of a 5-HT2 antagonist, ketanserin (KT) on vein graft morphology and function at 14 and 28 days after grafting. Forty-three New Zealand White rabbits underwent common carotid interposition bypass grafting. Twenty-two were treated with KT (0.86 mg/kg/day po) 5 days prior to surgery and thereafter until harvest. The remaining 21 animals acted as controls. VG were harvested at 14 (VG14) and 28 (VG28) days for histology or vasoreactivity. Twenty-three VG were harvested by pressure fixation and the midportions of the grafts were examined by videomorphometry. Standard isometric tension studies in response to serotonin and norepinephrine (NE) were performed on the rings from the remaining 20 VG. Contralateral external jugular veins (CV) from both groups were studied to assess the functional toxicity of KT. There were no toxic effects to KT noted. KT therapy did not affect the functional activity of the CV at 14 or 28 days when compared to controls. When compared to controls, KT significantly reduced the intimal thickness (49 +/- 11 vs 113 +/- 24 microns; P = 0.04) and there was an increase in luminal area (16.89 +/- 2.13 vs 8.41 +/- 1.59 mm2; P < 0.02) in VG28 only.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412063 TI - Precursor trapping: a "neonatal" mechanism of myocardial protection. AB - During induced ischemia for cardiac surgery, 5'-nucleotidase (5NT) catalyzes nucleotide breakdown by dephosphorylating AMP and IMP to diffusible precursors- adenosine and inosine. These precursors become unavailable upon reperfusion washout limiting nucleotide resynthesis, resulting in poor postischemic function. Neonatal hearts, which are more resistant to ischemia than adults, have low 5NT activity, trapping available precursors. Adult rabbit hearts given cardioplegia with a 5NT inhibitor, pentoxifylline, demonstrated improved postischemic contractility, compliance, and myocardial oxygen consumption after 120 min of 34 degrees C ischemia. To determine if this improved function was a result of enhanced nucleotide precursor availability during or following ischemia, total nondiffusible nucleotides, ATP, ADP, AMP, and IMP, and total diffusible nucleotides, adenosine, inosine, hypoxanthine, and xanthine, were measured by HPLC at end ischemia, 1 and 15 min after reperfusion. While all preischemic values were equivalent, pentoxifylline-treated hearts had significantly greater total non-diffusible nucleotides at end ischemia, 1 and 15 min after reperfusion. Additionally, pentoxifylline-treated hearts had significantly greater total diffusible nucleosides at end ischemia and 1 min after reperfusion, but were equal to control at 15 min after reperfusion. Furthermore, coronary sinus effluent had a significantly higher release of total diffusible nucleosides in control vs pentoxifylline-treated hearts. The data indicate that precursor trapping with pentoxifylline prevented nucleotide catabolism to diffusible precursors and enhanced postischemic nucleotide availability. We postulate the increased precursor availability augmented myocardial nucleotide resynthesis and correlated with the improved functional recovery noted. This strategy may have application in adult cardiac surgery. PMID- 8412064 TI - Regional geometry and function during myocardial ischemia and recovery. AB - To define the effects of altered left ventricular (LV) geometry on regional myocardial function during ischemia and recovery, regional and global LV geometry and transmural pressure (P) were measured in seven conscious dogs with sonomicrometry and micromanometry. Data were obtained at steady state and during rapid vena caval occlusion (VCO) under control conditions, after 15 min of left anterior descending occlusion, and after 1, 4, and 24 hr of reperfusion. Regional midwall minor axis (MA) Lagrangian strain (epsilon) and stress (sigma) were calculated from measured MA segment length (L), MA midwall radius, and wall thickness. Unstressed regional geometry was quantified using L0, the value of L at P = 0 during maximal VCO. Conventional (SWL) and normalized (SW sigma epsilon) regional MA stroke work were calculated for each cardiac cycle as the area of P vs L and sigma vs epsilon relationships, respectively. Regional Frank-Starling mechanisms corrected for changes in unstressed LV geometry were quantified as the slope (M sigma epsilon) of the linear end diastolic epsilon vs SW sigma epsilon relationship for data obtained during VCO (mean r = 0.98). M sigma epsilon returned to baseline levels within 1 h of reperfusion (P = 0.314 vs control). In contrast, 15 min of ischemia increased L0 by 15.2 +/- 2.5% (P < 0.05), which remained increased 5.7 +/- 1.7% above control values after 1 hr of reperfusion (P < 0.05). Both steady-state SWL and SW sigma epsilon decreased with ischemia and slowly returned towards baseline, remaining 28.7 +/- 7.5% and 26.4 +/- 6.3% below control values after 1 hr of reperfusion (both P < 0.05). Therefore, late functional recovery from reversible ischemic injury is primarily correlated with reversal of changes in regional geometry, specifically the reversal of diastolic creep. As a result, adequate quantification of postischemic regional myocardial performance requires characterization of changes in regional geometry as well as indicators of Frank-Starling mechanisms. PMID- 8412065 TI - Intracellular glutamine concentration does not decrease in all muscles during sepsis. AB - The concentrations of glutamine and other amino acids were measured in plasma and intracellular fluid of soleus and extensor digitorum longus (EDL) muscles of rats 4, 8, and 16 hr after induction of sepsis by cecal ligation and puncture or after sham operation. Previous studies have shown that muscle protein breakdown is greatly increased in EDL, but not in soleus muscle, in this sepsis model. Corresponding to previous observations of protein breakdown in sepsis, muscle glutamine was markedly depleted (< 50%) in EDL by sepsis, while no significant fall in glutamine concentration in soleus was observed. Changes in muscle glutamine concentration in sepsis could not be attributed to changes in the precursor of glutamine, glutamic acid. Data were examined for changes consistent with hypothesized alterations in glutamine transport. Correlations among glutamine and other amino acids in muscle, histidine in particular, were consistent with a sepsis-induced alteration in activity of the sarcolemmal glutamine transporter, system Nm. These results thus strengthen the proposed connection between muscle glutamine content and muscle protein metabolism under catabolic conditions. PMID- 8412066 TI - Lymphokine activated killer cells enhance IL-2 prevention of sepsis-related death in a murine model of thermal injury. AB - It has previously been shown by this laboratory that immunomodulation of thermally injured animals with low-dose interleukin-1 (IL-2) and indomethacin (Indo) improves survival following septic challenge. Lymphokine-activated killer (LAK) cells have been shown to be effective in certain viral infections and to act in synergy with IL-2 in the treatment of certain types of cancer. We have studied the effect of LAK cells in combination with IL-2 and Indo in a murine model of thermal injury and sepsis. Male A/J mice received a 25% scald burn injury or sham burn and were randomized into five groups: (a) sham/vehicle, (b) burn/vehicle, (c) burn/IL-2 (250 U) + Indo (5 micrograms), (d) burn/LAK cells (2 x 10(6) cells), or (e) burn/LAK cells+IL-2+Indo and were treated accordingly for 6 days following injury. LAK cells were generated by in vitro IL-2 treatment of syngeneic spleen cells for 72 hr and cytotoxic activity was confirmed by standard 51Cr release assay using natural killer (NK)-sensitive and NK-resistant targets. In the groups receiving LAK cells they were administered on Day 1 and Day 6 postinjury. On Day 10, septic challenge by cecal ligation and puncture (CLP) or splenectomy, for in vitro studies, was performed. Five-day survival after CLP was 80% in the sham/vehicle group compared to 0% in the burn/vehicle group (P < 0.01). IL-2/Indo and LAK/IL-2/Indo improved survival to 25% (P < 0.05) and 57.1% (P < 0.01), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412067 TI - Demonstration of interstitial collagenase in abdominal aortic aneurysm disease. AB - This study was performed to evaluate the presence of interstitial collagenase, now known as matrix metalloproteinase-1 (MMP-1), in specimens of abdominal aortic aneurysms (AAA). Eight AAA and four control infrarenal aortas were evaluated. After homogenization and extraction of soluble proteins, immunoblots of the extracts equalized for protein content were performed with a specific antibody to MMP-1. Under native conditions, immunoreactive material was distributed between M(r) 27 kDa to > 106 kDa. When the extracts were reduced and denatured, immunoreactive bands were detected in AAA at the expected M(r)'s of the secreted isoforms (57 and 52 kDa), whereas control aortic extracts had low levels of detectable immunoreactive material. Only extracts from AAA demonstrated significant immunoreactivity to the lower M(r) isoforms (22, 25, and 27 kDa), which correspond to reported cleavage products of MMP-1. Preliminary immunofluorescent studies of AAA localized MMP-1 to cells present in the adventitia of AAA. These findings will help to resolve disagreement in the recent literature regarding the presence of collagenolytic activity in AAA disease. PMID- 8412068 TI - Intestinal production of interleukin-1 alpha during endotoxemia in the mouse. AB - Interleukin-1 (IL-1) may be involved in gut permeability to macromolecules and gut glutamine metabolism during endotoxemia. We developed a sensitive radioimmunoassay specific for mouse IL-1 alpha (detection limit of 100 pg/ml, or 5 pM) and measured intestinal levels of IL-1 alpha in response to endotoxin. CD-1 mice (N = 190) were randomized to intraperitoneal (ip) or intravenous (i.v.) lipopolysaccharide (LPS) infusion (15 micrograms/g or 1.5 micrograms/g Escherichia coli 0111:B4 LPS) or saline. Mice were sacrificed at Time 0, 30 min, 1 hr, 2.5 hr, 4 hr, 6 hr, 12 hr, and 24 hr (3 mice/group/time point). Small bowel (SB) and large bowel (LB) were harvested and compared to liver. Duodenum, upper jejunum, midjejunum, terminal ileum, cecum, ascending colon, and sigmoid were analyzed in separate experiments. Tissues were frozen, weighed, and homogenized, the homogenates were centrifuged, and the supernates were assayed for immunoreactive IL-1 alpha. IL-1 alpha was expressed as pg/g wt +/- SEM (lowest detectable amount = 1000 pg/g wet tissue (WT)). SB but not LB from normal controls had constitutively elevated levels of IL-1 alpha (6177 +/- 1640 pg/g WT). LPS ip or i.v. produced lethargy, diarrhea, and a dramatic elevation of IL-1 alpha levels in both SB and LB. In SB, IL-1 alpha was elevated compared to baseline at 1 hr (19201 +/- 626 pg/g WT) and reached a fivefold maximal increase at 2.5 hr (31775 +/- 503 pg/g WT) following 15 micrograms/g ip.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412069 TI - Dichloroacetate inhibits peripheral efflux of pyruvate and alanine during hormonally simulated catabolic stress. AB - The purpose of this study was to examine the relationship of peripheral metabolism of glucose, lactate, alanine, and muscle protein catabolism to pyruvate availability during stress. Peripheral catabolic stress was simulated by the infusion of epinephrine, cortisol, and glucagon into the femoral artery of 12 healthy volunteers, 6 of whom received prior treatment with dichloroacetate while 6 served as controls. The catabolic hormone infusion reproduced the peripheral stress response in which glucose consumption increased and the efflux of lactate, alanine, and total amino acid nitrogen (i.e., net muscle protein catabolism) from the leg increased. Dichloroacetate (DCA), which is known to increase pyruvate oxidation, reduced the hormonally stimulated efflux of pyruvate and alanine from the leg and decreased the rate of extremity glucose consumption. DCA had no effect on the rate of lactate efflux and except for alanine had no effect on the stimulated rate of total amino acid nitrogen loss. These results demonstrate the dependence of alanine efflux on pyruvate availability during stress and suggest that the rate of glycolysis within peripheral tissues is a major factor in regulating the quantity of alanine efflux. This study further illustrates that except for alanine, pyruvate kinetics are not salient in the regulation of muscle protein catabolism and elucidates the dichotomy between alanine kinetics and true muscle protein breakdown. PMID- 8412070 TI - Decreased oxidized glutathione with aerosolized cyclosporine delivery. AB - Cyclosporine immunosuppression remains vital for successful lung transplantation. Cyclosporine also functions as a membrane active biological response modifier and has been noted to have a variable effect on ischemia-reperfusion (I/R) injury in various tissues. Glutathione plays an important role in the endogenous antioxidant defense system; plasma oxidized glutathione (GSSG) levels are useful as a sensitive indicator of in vivo oxidant stress and I/R injury. Lung transplantation results in ischemia, followed by a period of reperfusion, potentially producing functional injury. This study was designed to evaluate the effect of cyclosporine on oxygen radical generation in a model of single-lung transplantation. Single-lung transplantation was performed in 12 mongrel puppies, with animals assigned to receive either intravenous or aerosolized cyclosporine. Arterial blood and bronchoalveolar lavage fluid (BALF) samples were obtained to determine GSSG levels via a spectrophotometric technique. Samples were obtained both prior to and following the revascularization of the transplanted lung. Whole blood and tissue cyclosporine levels were determined via an high-performance liquid chromatography technique 3 hr following the completion of the transplant. Aerosolized cyclosporine administration resulted in greatly decreased arterial plasma and BALF GSSG levels, whole blood cyclosporine levels, and equivalent tissue cyclosporine levels when compared to intravenous cyclosporine delivery. These findings support the hypothesis that the transplanted lung is a source of GSSG production and release into plasma. Additionally, these findings suggest that cyclosporine may have a direct antioxidant effect on pulmonary tissue, with this activity occurring at the epithelial surface, an area susceptible to oxidant injury. PMID- 8412071 TI - Luminal adrenergic agonists modulate ileal transport by a local mechanism. AB - Intestinal transport is controlled by neural pathways, hormones, and luminal agents. Luminal adrenergic agents influence water and ion transport in the jejunum. This study tested two hypotheses: (i) luminal adrenergic agents influence ileal water, ion and glucose transport, and (ii) luminal adrenergic agents exert their effects locally and selectively. Absorption studies (n = 46) were performed on dogs with two adjacent 25-cm ileal Thiry-Vella fistulas (TVF). Perfusion with [14C]polyethylene glycol was used to calculate absorption of water, ions, and glucose from the distal TVF. Experiments were composed of three 1-hr periods: basal, luminal adrenergic agonist infusion, and recovery. In group 1 the adrenergic agonists were administered to the distal TVF: norepinephrine (alpha 1 > alpha 2 and beta), phenylephrine (alpha 1), clonidine (alpha 2), and isoproterenol (beta). In group 2 the adrenergic agonists were administered to the proximal TVF, with absorption measured in the distal TVF. In group 1 norepinephrine and phenylephrine caused a significant increase in water absorption (P < 0.05). Clonidine and isoproterenol caused decreased absorption of water and ions, with clonidine causing significantly decreased absorption (P < 0.05) of water, ions, and glucose. In group 2 there were no changes in distal TVF absorption. Luminal adrenergic agents did not alter the heart rate in either group. Luminal adrenergic agonists modulate ileal transport via a local mechanism. A proabsorptive response is observed with alpha 1 agonists, while alpha 2 and beta agonists cause a prosecretory effect. Inhibition of glucose absorption appears to be selectively mediated via the alpha 2-adrenergic receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412072 TI - Reversible contractile dysfunction in reversible experimental cardiac allograft rejection: alterations in the beta-receptor-stimulated adenylylcyclase pathway. AB - Alterations in the beta-adrenergic receptor adenylylcyclase (AC) pathway are well known in heart failure. Previous studies by our group have demonstrated impaired function of the AC pathway and contractile reserve when stimulated with isoproterenol (ISO) or forskolin (F) in cardiac allografts with moderate or severe rejection. To determine if recovery of the AC pathway occurs when rejection is reversed, we used a rat heterotopic heart transplant model. Lewis (L) rats received either isografts or Lewis-Brown Norway (LBN) allografts (ALLO). The hearts were explanted on Day 4 and reimplanted into a recipient syngeneic with the original donor (L to L, LBN to LBN). Grafts were harvested 2 days later and analyzed. Receptor-mediated modulation of AC activity was investigated using ISO, F, and the metabolic inhibitor of AC, R-N6-(2-phenylisopropyl)-adenosine (R PIA). ISO- and F-mediated stimulation of AC were significantly impaired in ALLO (see Table). CARB and R-PIA remained unchanged. [table: see text] Reimplanted hearts had no histologic evidence of rejection and had normal cAMP production in response to ISO and F. In conclusion, AC alterations are concordant with histologic changes in this reversible model of heart failure. Such alterations may be a component in the contractile dysfunction associated with rejection. PMID- 8412073 TI - Identification of dopamine1A receptors in the rat small intestine. AB - Dopamine receptors have been localized to several tissues outside the central nervous system including the kidney and mesenteric vessels. To determine if there are dopamine1 receptors within the small intestine, homogenates of the antimesenteric halves of the entire jejunum and ileum of adult rats were prepared and competitive inhibition studies and Scatchard analysis were performed at room temperature using 125I-SCH 23982 and SCH 23390. The specific binding of 125I-SCH 23982 to the intestinal tissue homogenates was rapid, saturable with ligand concentration, and reversible. Analysis of the Scatchard plots revealed a single class of receptors with an apparent dissociation constant of 10.77 +/- 2.32 nM and maximum receptor density of 1.37 +/- 0.34 fmole/mg protein. Emulsion autoradiography performed using 125I-SCH 23982 on antimesenteric sections of the rat small intestine revealed that the dopamine1 receptors are located on cells at the base of the intestinal crypts. Two dopamine1 subtypes (D1A and D1B) have been identified by molecular biological techniques. Using a ribonuclease protection assay we found expression of the D1A receptor gene in the small intestinal tissue. These studies are the first to identify, characterize, and localize receptors for the endogenous catecholamine, dopamine, within the rat small intestine and to confirm the expression of the D1A receptor gene. PMID- 8412074 TI - Fibroblast transplantation in rats: transduction and function of foreign genes. AB - The purpose of this study was to develop a method of transduction of normal skin fibroblasts with a retroviral vector expressing the human factor IX (hF-IX) gene and transplantation of the transduced cells into rats. A retroviral vector containing the hF-IX gene, as well as a selectable marker (hygromycin B resistance; HB), was used to transduce cultured normal skin (rat and human) fibroblasts. Transduced fibroblasts were selected with HB; resistant clones were propagated and assessed for expression of active hF-IX in vitro. Transduced cells secreted significant amounts of hF-IX in vitro (327.9 +/- 14.2 ng/10(6) cells) which showed measurable clotting activity, indicating terminal processing of the gene-transcribed hF-IX product by the transduced fibroblasts. Integration of the proviral gene into rat and human fibroblast genomic DNA was confirmed by polymerase chain reaction. Transduced rat fibroblasts were attached to collagen coated microcarriers and transplanted intraperitoneally into syngeneic recipients. Transplanted microcarrier-attached cells formed aggregates in the peritoneal cavity and exhibited positive immunohistochemical staining for hF-IX up to 8 weeks following transplantation, the time of termination of the experiment. These experimental results may have significant implications in developing strategies for future clinical therapy for hemophilia B. PMID- 8412075 TI - Endogenous epidermal growth factor regulates limb development. AB - Mutations associated with genes of the EGF superfamily are implicated in limb malformations. To evaluate the potential role of EGF-mediated signal transduction in the control of early mammalian limb development, we developed a simple in vitro system which is permissive for morphogenesis and cytodifferentiation in serumless, chemically defined medium. Our experimental strategy was to ascertain if the EGF precursor gene was transcribed and translated into potentially bioactive growth factor. EGF mRNA transcripts are expressed in Swiss Webster mouse embryonic (42-44 somite pairs) forelimbs as determined by mRNA phenotyping. EGF transcripts are translated into precursor EGF polypeptides which were localized to limb covering epithelium and the chondrogenic mesenchymal cell lineages. EGF immunostaining patterns suggested a paracrine type of regulation for the cartilage blastema associated with forelimb development. To test whether EGF effects the timing and positional information required for limb-specific cartilage morphogenesis, we employed tyrphostin (RG 50864) which inhibits EGF receptor kinase activity in a concentration-dependent manner and severely retards limb development. These findings support our hypothesis that endogenous EGF or EGF-like proteins provide signaling for the size and shape of discrete forelimb cartilage formations during mouse embryonic morphogenesis. PMID- 8412076 TI - Selective management of iatrogenic femoral artery injuries. AB - Increased femoral artery catheterization for therapy and diagnosis has resulted in a concomitant increase in iatrogenic femoral artery injuries. A 33-month experience at two affiliated institutions was reviewed to evaluate the results of a selective approach to management of these complications. During this period, 9576 femoral artery catheterizations were performed. Eighty-nine femoral artery injuries were treated surgically or by ultrasound-guided compression therapy. Cardiac catheterization procedures predominated and 61% of patients were anticoagulated. Thirty-eight cases were treated surgically: 14 pseudoaneurysms, 16 hematomas, 6 arteriovenous fistulas, 2 thromboses. Local anesthesia was used in 28 cases (74%). Arterial repair was required in 34 cases, with control being achieved below the inguinal ligament in 33 cases (97%). Punctures were found in the superficial femoral or profunda femoral rather than the common femoral artery in 17 of 38 cases (44%, P < 0.001 compared with the ultrasound group). There were two deaths (5.3%), six wound infections (16%), and no limb loss. Ultrasound guided compression was preferentially used for stable or slowly expanding pseudoaneurysms. Cases with large hematomas causing skin ischemia were treated surgically. The pseudoaneurysms ranged from 2 to 7 cm in diameter. This technique was effective in 46 of 51 cases (90%). A single thromboembolic complication was treated by thrombolysis. There were no late recurrences. We conclude that (1) iatrogenic femoral artery injuries are associated with postcatheterization anticoagulation and punctures not located in the common femoral artery; (2) injuries requiring surgery can usually be treated under local anesthesia with infrainguinal arterial control; (3) ultrasound-guided compression is an effective method for treating iatrogenic pseudoaneurysms not associated with large hematomas. PMID- 8412077 TI - Portal endotoxemia stimulates the release of an immunosuppressive factor from alveolar and splenic macrophages. AB - Impairment of cell-mediated immunity is both a common manifestation of critical illness and a potential cause of increased infectious morbidity and mortality. The mechanisms responsible for alterations in systemic immune regulation are incompletely understood; however, monocytes and fixed tissue macrophages appear to play a central role. We have previously shown that infusion of gram-negative organisms into the portal vein, but not into the systemic circulation, induces suppression of delayed hypersensitivity responsiveness in vivo and of mitogen stimulated lymphocyte proliferation in vitro. The present studies were undertaken to probe the mechanisms of this suppression. Rats received 3 x 10(8) killed Pseudomonas aeruginosa via the inferior vena cava or the portal vein; they were sacrificed 24 hr later and the mitogen-driven proliferative responses of isolated splenocytes were assayed. Portal infusion resulted in significant suppression of Con A-induced proliferative responses (15.5 +/- 2.7 cpm x 10(-3) compared to 68.6 +/- 9.8 cpm x 10(-3) for infrahepatic vena cava-infused animals and 48.0 +/- 5.4 cpm x 10(-3) for nonoperated controls). Suppression was shown to be a consequence of the release of a soluble suppressive factor from splenic adherent cells. Suppression of the proliferative responses of control lymphocytes could also be induced by a soluble factor present in culture supernatants of alveolar macrophages harvested from portally infused animals (4.7 +/- 0.4 cpm x 10(-3) vs 88.6 +/- 27 cpm x 10(-3) for systemically infused animals and 60.1 +/- 8.4 cpm x 10(-3) for nonoperated controls). The stimulus for the release of this factor was not endotoxin, but a second factor released from the liver.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412078 TI - Panmyelic epidural cooling protects against ischemic spinal cord damage. AB - The neuroprotective effect of epidural cooling before and during spinal cord ischemia on the neurological, neurophysiological, and histopathological outcome was evaluated after 40 min of proximal and distal thoracic aorta crossclamping in dogs. In the normothermic group (n = 12), no attempt was made to change the spinal cord temperature. Four of eight animals showed complete paraplegia and four had partial recovery. The N3 component of spinal somatosensory-evoked potentials recovered to only 11.7 +/- 1.4% after 2 hr of recirculation and to 45% of control value after 2 days of survival. In the transverse sections taken from L1-L7 segments, apparent interneuronal damage in the intermediate zone was found after 2 hr of reperfusion followed by a heavy loss of interneurons after 2 days of survival and functionally defined as fully developed paraplegia. In the hypothermic group (n = 12), the spinal cord temperature was lowered 3 min before aortic crossclamping with a bolus of epidurally administered 0.9 N saline solution (8 ml/kg at 5 degrees C) to 28.5 +/- 1.3 degrees C and was maintained throughout the crossclamping time with the additional infusion of the same solution (20 ml/kg/40 min) using a peristaltic pump. Seven of eight animals had no neurological deficit and one animal showed partial recovery, which was significantly better than the motor score for the normothermic group (P < 0.05). The SSEP revealed 55% of postsynaptic (N3) wave recovery after 2 hr of recirculation and 92% recovery after 2 days survival, which was significantly higher than those for the normothermic animals (P < 0.05). Histological analysis showed almost full protection of interneurons and A-motoneurons verified after 2 hr and 2 days, respectively. We conclude that spinal cord epidural cooling has a highly protective effect against ischemic spinal cord damage under experimental conditions of high thoracic aorta crossclamping in dogs. PMID- 8412079 TI - Double-layer prostheses for repair of abdominal wall defects in a rabbit model. AB - The primary objective of this study was to compare the effectiveness of expanded polytetrafluoroethylene (e-PTFE) and polypropylene (PPM) in a rabbit abdominal wall defect model using a double-layer repair technique. Fifty-four New Zealand white rabbits had full thickness resection of a 5 x 5-cm segment of the mid portion of at the abdominal wall. The defect was repaired with two 6 x 6-cm layers of prosthesis in an underlay/overlay fashion incorporating a 1-cm edge of abdominal wall and grouped accordingly. Group I: inner and outer layer of PPM; Group II: inner layer e-PTFE, outer layer PPM; Group III: inner and outer layer e PTFE. Animals from each group were sacrificed at 3, 6, and 12 months. The abdominal wall was assessed for herniations and bowel adhesions to the inner layer of the prosthesis. Adhesions were graded according to an adhesion scoring system (grade 0-3). No herniations were observed. Intestinal adhesions to PPM were quite dense at 3 months with a mean adhesion score of 2.5; at 12 months, adhesions were more dense and extensive, having a mean score of 3. Adhesions to e PTFE were thin and filmy with a mean score of 0.4 at 3 months and 0.6 at 12 months (P < 0.001, 95% CI for each time period). Histologically, PPM fibers were consistently surrounded by dense fibrous connective tissue; foreign body giant cells were present. A thin connective tissue capsule covered the inner e-PTFE layer; its pores were progressively infiltrated by fibroblasts. The double-layer technique prevented hernias in all groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412080 TI - Localization of sarcoma xenografts in nude mice with indium-111-labeled monoclonal antibodies. AB - A labeling method utilizing modified carbohydrate moieties in antibody heavy chains as radionucleotide binding sites was evaluated. Murine anti-sarcoma monoclonal antibody (MAb 19-24) was labeled with Indium-111 (111In) using this technique and subcutaneous human sarcoma xenografts were successfully localized in nude mice. A nonspecific monoclonal antibody BL-3 was used as a negative control. Tumor-to-blood ratios of radioactivity in the mice injected with 111In labeled MAb 19-24 were significantly (P < 0.05) higher than those obtained with nonspecific MAb BL-3. Calculations of percentage injected dose of radioactivity per gram tissue showed relatively high specific uptake of MAb 19-24 in sarcoma xenografts. Radioactivity cleared from the blood rapidly and hepatic uptake of 111In-labeled antibodies was found to be relatively low. Biodistribution studies in normal mice with 111In-labeled antibodies showed only blood pool activity with no significant concentration of activity into organs. Therefore, immunoreactivity of the antibodies was retained after 111In-labeling utilizing this new technique, allowing specific binding of radiolabeled MAb to tumor xenografts with relatively low hepatic uptake. PMID- 8412081 TI - Hormonal and splanchnic hemodynamic alterations following hepatic resection. AB - Reported hemodynamic changes after partial hepatectomy (PH) include elevations in portal blood flow and pressure (Ppv). The etiology of these changes is unknown. We studied degree of portosystemic shunt (PSS), splanchnic artery blood flow and resistance (Qspl, Rspl) as well as prostacyclin (PGI2), glucagon, and insulin response in rats undergoing 75% hepatectomy. The results showed that following an immediate threefold rise in portal PGI2, a hemodynamically significant PSS developed at 48 hr. The PGI2 changes paralleled the increase in portal flow (Qpv), pressure (Ppv), PSS and the decrease in Rspl. Glucagon levels also rose immediately postresection, but fell to normal prior to resolution of both PSS and hyperemia. There was no significant change in insulin or glucose levels after resection. This implies a relationship between PSS and the release of a prostanoid effecting Qpv, Qspl, and Rspl, which relates to, and possibly mediates, the hemodynamic changes following liver resection. We conclude that PH is a model of acute PHT, and that the altered splanchnic hemodynamic could relate to hepatic regeneration. PMID- 8412082 TI - A 5-HT3 receptor agonist induces neurally mediated chloride transport in rat distal colon. AB - Having previously demonstrated that serotonin (5-HT)-induced chloride secretion in rat distal colon is mediated at both neural and nonneural receptors, we isolated the neural component of this response by adding the selective 5-HT3 receptor agonist, 2-methyl-5-hydroxytryptamine (2Me5HT), to in vitro sheets of rat distal colon with intact neural plexuses. Rats were sacrificed, and the distal colon excised, opened, cut into sections and mounted, all layers intact, as flat sheets in Ussing chambers under short-circuited conditions. 2Me5HT induced a prompt, significant (P < 0.01), concentration-dependent rise in short circuit current (Isc; EC50 6.2 microM); 50 microM 2Me5HT decreased both net sodium and chloride absorption (-0.1 +/- 0.5 and -2.1 +/- 0.8 muEq/cm2 x hr, respectively); the difference (2.0 +/- 0.8 muEq/cm2 x hr) in these changes was not statistically different from the rise in Isc (1.5 +/- 0.3 muEq/cm2 x hr). Since the only significant change in unidirectional flux was the rise in electrogenic Cl- secretion (P < 0.01), the delta Isc induced by 2Me5HT may be used as a measure of electrogenic chloride secretion induced by the agonist. The rise in Isc induced by 2Me5HT was abolished by both 0.2 microM tetrodotoxin and 0.1 microM ICS 205-930 (a 5-HT3 antagonist) but was not inhibited by 1.0 M atropine 100 microM hexamethonium, 10 microM phentolamine, 10 microM propranolol, 10 microM 5-HTP-DP (a 5-HT1P antagonist), or 0.1 microM ketanserin (a 5-HT2 antagonist). These results indicate that 2-methyl-5-HT is a highly selective agonist for neurally based 5-HT3 receptors in this model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412083 TI - Hepatic neutrophil influx: eicosanoid and superoxide formation in endotoxemia. AB - We examined early changes, at 0.5, 1.5, and 3 hr of infusion of a nonlethal dose of Escherichia coli endotoxin (ET) in neutrophil (PMN) sequestration in the liver and accompanying alterations in [1-14C]arachidonic acid metabolism and superoxide anion release by Kupffer and endothelial cells and PMNs. One hundred fifty micrograms ET (268 micrograms/kg) was infused over 3 hr. Shorter infusions delivered proportionally less ET. By 30 min of ET infusion the number of PMNs in the 45 ml/min fraction was 2.41 x 10(7), eightfold higher than that in the NaCl infused rats, representing 35.90 +/- 3.49% (n = 7) of the total cell yield vs 8.20 +/- 0.20% (n = 4) in NaCl controls. By 90 and 180 min of ET infusion the number of PMNs and their share of the total cell yield increased significantly further. Cells were separated by elutriation, followed by application of a Ficoll Hypaque gradient, when appropriate. Kupffer cells and PMNs were recovered in the 45 ml/min fraction, endothelial cells in the 23 ml/min fraction. At 30 min of ET infusion the profile of arachidonic acid metabolites released from [1 14C]arachidonic acid-prelabeled nonparenchymal cells upon A23187 stimulation was not different from that of cells of NaCl-infused rats. Infusion of ET for 90 min accentuated PMN infiltration, and resulted in significant modulation of the eicosanoid profile, consisting of a major shift in PGD2/PGE2 to PGE2, and LTB4 and 12-HETE accounting for 34% of the total eicosanoids, compared to 12.9% in time-matched NaCL controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412084 TI - Effect of tumor burden on futile glucose and lipid cycling in tumor-bearing animals. AB - Futile cycling of metabolic substrates may play an important role in the development of cancer cachexia. To determine the effect of tumor burden on futile glucose and lipid cycling, 64 female Lewis/Wistar rats were randomized to control (non-tumor-bearing), small tumor burden, or large tumor burden groups (MAC-33 mammary adenocarcinoma). After 5 or 25 days of tumor growth, animals received a 5 day period of parenteral nutrition (158 kcal/kg/day) followed by a 6-hr infusion of the stable isotopes glucose-2-d and glucose-6,6-d2 or [C13]palmitate. The heavy glucose isotopes glucose-2-d and glucose-6,6-d2 are labeled with deuterium at the 2-carbon position and doubly labeled with deuterium at the 6-carbon position, respectively, to obtain differential molecular weights. No increase in glucose or lipid cycling was observed in animals with small tumor burdens. In contrast, a significant increase in plasma rate of appearance (Ra) of glucose-2-d (1377 +/- 136 mg/hr vs 947 +/- 54 mg/hr), Ra of glucose-6,6-d2 (810 +/- 88 mg/hr vs 510 +/- 24 mg/hr), and total glucose cycling (548 +/- 57 mg/hr vs 416 +/- 28 mg/hr) was seen in animals with large tumor burdens compared to control animals (P < 0.05). Although a trend toward increased lipid cycling was seen in tumor bearing versus control animals, this change was not significant. Thus, futile cycling of glucose was significantly elevated in animals with large tumor burdens and may cause significant energy wasting to contribute to the development of cachexia in the tumor-bearing host. PMID- 8412085 TI - Inhibition of tumor metastasis by a circulating suppressor factor. AB - Increased growth of tumor metastases has been reported after primary tumor excision in animal models. Postulated mechanisms for this phenomenon include circulating suppressor factors secreted by the primary tumor, tumor manipulation during surgical excision, and limiting nutritional factors which are consumed by both primary and metastatic tumors. To study this phenomenon, Lewis/Wistar rats with subcutaneous mammary adenocarcinoma implants (MAC-33) were randomized to receive either a standard protein diet (22.0% protein, 4.20 kcal/g) or a protein depleted diet (0.03% protein, 4.27 kcal/g) ad lib per os. Twenty-one days after tumor inoculation, half of the animals in each dietary group underwent primary tumor excision. Control animals underwent sham excision on the flank contralateral to the tumor and physical manipulation of the primary tumor. At sacrifice 35 days after tumor inoculation, a significant increase in regional metastasis (axillary lymph nodes) and distant metastasis (lungs) occurred after primary tumor excision in animals receiving the standard protein diet. No increase in regional or distant metastases were found in animals receiving the protein-depleted diet after tumor excision. Serum was collected from animals given the standard protein diet following tumor or sham excision and added to MAC 33 tumor cell cultures for in vitro determination of tumor cell proliferation. [H3]Thymidine incorporation by MAC-33 cells in vitro was significantly suppressed by serum from tumor-bearing animals. These results imply that the biologic mechanism mediating this phenomenon is a circulating suppressor factor of tumor metastasis which is produced in tumor-bearing animals receiving standard protein (but not protein-depleted) diets. PMID- 8412086 TI - Effect of intraperitoneal prosthetic materials on reticuloendothelial function in the rat. AB - The effect of different intraperitoneal prosthetic biomaterials on reticuloendothelial system (RES) function and bacterial translocation were studied in the rat. Rubber drains, knitted dacron (KD), or silicone elastomer (SE) with surface areas of either 3 or 10 cm2 were implanted into the lower right part of the abdominal cavity under aseptic conditions. RES function, expressed as the phagocytic index and the uptake of 125I-labeled Escherichia coli (cpm/g tissue) in systemic organs (liver, spleen, lungs, and kidneys) and in gut associated lymphoid tissues [GALT; i.e., mesenteric lymph nodes (MLN), jejunum, ileum, and colon], was measured 1 day after implantation/sham operation. A significant elevation of the phagocytic index was noted in all implanted groups, except the group with 3 cm2 of SE, as compared with controls. The uptake of 125I labeled E. coli significantly increased in the liver (rubber, 10 cm2), spleen (rubber, 10 cm2), lungs (rubber, 10 cm2; KD, 3 and 10 cm2; SE, 3 and 10 cm2) and kidneys (rubber, 10 cm2), but significantly decreased in the MLN and jejunum in all implanted groups, in ileum (KD, 10 cm2; SE, 3 cm2), and colon (rubber, 10 cm2; KD, 3 and 10 cm2; SE, 3 cm2). The percentage uptake significantly increased in the systemic organs and decreased in the GALT in most of the implanted groups. Indigenous bacterial translocation to the MLN, liver, spleen, lungs, and kidneys occurred simultaneously. A size-dependent fashion in influence on RES function and incidence of bacterial translocation was observed. The results suggest that intraperitoneal prosthetic biomaterials alter host RES function with concomitant bacterial translocation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412087 TI - Cardiac contractile and calcium transport function after burn injury in adult and aged guinea pigs. AB - Cardiac dysfunction occurs after a major burn injury regardless of age; whether burn trauma causes greater myocardial contractile depression in the older subject due to reduced cardiac reserves that normally occur with adult aging is not known. The cellular basis for burn-induced cardiac dysfunction is not known, but several studies have suggested that alterations occur in the rate of Ca2+ delivery to the contractile proteins as well as in the rate of Ca2+ removal from the sarcoplasm by the sarcoplasmic reticulum (SR). To determine if age-related differences in the cardiac contractile response to burn injury are associated with differences in SR Ca2+ transport, an isolated heart preparation was used to examine mechanical function, and myocardial homogenate preparations were used to assess SR Ca2+ transport. Guinea pigs from both age groups (adult, 6-8 months, and senescent, 34-36 months of age) were divided into two subgroups--control and 45% cutaneous scald burn. Cardiac dysfunction associated with adult aging alone was indicated by lower systolic pressure and lower rates of left ventricular (LV) pressure rise and fall, as well as decreased responses to isoproterenol, exogenous Ca2+, increased coronary flow rate, and electrical pacing. Myocardial depression in senescent control hearts was accompanied by a decreased maximal Ca2+ uptake in myocardial homogenates, suggesting that altered SR calcium transport may contribute to the diminished contractile function associated with aging. Burn injury impaired cardiac function in all animals regardless of age as evidenced by a leftward shift of LV function curves and altered responses to receptor- and nonreceptor-mediated inotropic interventions. However, the percentage change in cardiac function after burn injury was similar in both age groups compared to those of their respective controls. Significant alterations in SR Ca2+ transport were observed in myocardial homogenates isolated from both adult and senescent hearts after burn injury. Our data confirm that burn injury induced cardiac contractile dysfunction as well as alterations in SR Ca2+ transport function regardless of age, and we conclude that altered SR Ca2+ transport function contributes, in part, to the diminished cardiac function after burn injury. PMID- 8412088 TI - In vivo deleterious effects of a right shift of the HbO2 curve during hypoxemia. AB - We investigated the influence of a right shift of the oxyhemoglobin dissociation curve on tissue oxygenation in two groups of anesthetized rabbits subjected to short periods of graded hypoxia: Group 1 (n = 5) with elevated P50 due to increased RBC 2,3-diphosphoglycerate and adenosine triphosphate and Group 2 (n = 5) with normal P50. Hemoglobin fell progressively in all animals due to blood letting for necessary measurements. During 16% inspired O2 (FIO2), both groups remained stable. During 13% FIO2, arterial pO2 was the same in both groups, but only in Group I did it fall below the crossover point (C.O.P.), which was raised by the high P50. Arterial pH and arterial-venous O2 content difference remained within the normal range in both groups throughout the experiment. During 13% FIO2, animals with high P50 showed a fall in cardiac output and oxygen consumption while animals with normal P50 remained stable. We postulate that when systemic O2 content is sufficiently reduced and tissue O2 extraction is maximal, the O2 needs of the myocardium perfused with a pO2 below the C.O.P. cannot be met: under these conditions cardiac output and systemic O2 consumption fall, presumably due to a reduction in coronary blood flow. PMID- 8412089 TI - In situ macrophage distribution in the hepatic allograft associated with rejection in the rat. AB - The association of macrophage infiltration with orthotopic rat liver transplant rejection was studied in the ACI(RT1a) to the LEW(RT1(1)) rat combination, using immunohistochemical staining with several monoclonal antibodies at different time points. LEW recipients of ACI liver transplants experienced severe acute rejection, with a mean survival of 10.2 +/- 0.7 days. An indirect immunoperoxidase technique on cryostat sections of the liver grafts was used to determine the localization of macrophages infiltrating the grafts as defined by specific rat anti-macrophage monoclonal antibodies designated TRPM-3 and KI-M2R. In addition, the monoclonal antibodies MRC OX3 and MRC OX6 were used to detect the macrophages that expressed Ia antigens. MRC OX3 binds only with the host LEW Ia antigens, but MRC OX6 binds with both host LEW and donor ACI Ia antigens. OX3+ and OX6+ macrophages were seen in the periportal area of the hepatic allograft on Day 5; thereafter, the number of these cells increased in the periportal and pericentral areas as well as in the sinusoidal lumens. A large number of TRPM-3+ cells were also seen at the periportal area in the hepatic allograft on Day 5, whereas there was no increase in the number of KI-M2R+ cells. In addition, most of the infiltrating mononuclear cells were OX3+. These findings suggest that a large number of TRPM-3+ macrophages of recipient origin migrate into the hepatic allograft soon after transplantation. A progressive relative increase in host TRPM-3+ macrophages is a characteristic feature of ongoing first-set rejection in the rat hepatic allograft. PMID- 8412090 TI - Use of the hydrogen clearance technique for measurements of pancreatic blood flow. AB - The hydrogen clearance technique (HCT) was employed for measurements of local pancreatic blood flow (PBF) in dogs, opossums, and rats under normal conditions and in opossums during biliary pancreatitis. Local PBF (ml/min/100 g) in dogs was 57.6 +/- 10.4, in opossums 60.6 +/- 9.5, and in rats 144.7 +/- 23.4, under resting conditions. Regional distribution of local PBF in dogs showed no statistical differences. Technical aspects of the HCT were described in detail. Under physiological conditions parenchymal trauma following electrode implantation was negligible in nearly all cases. More than 95% of registered clearance curves were monoexponential and showed excellent reproducibility of PBF values in repeated measurements (percentage deviation < or = 6.7). Five days after initiating biliary pancreatitis by common channel ligation in opossums local PBF was 72.1 +/- 14.7 after implantation of new electrodes. Chronically implanted electrodes, however, recorded only 39.7 +/- 15.8, due to massive fibro proliferative alterations around the implanted electrode tips. We therefore conclude that the HCT as described is a useful tool for measuring local PBF in acute experimental settings only. Biliary pancreatitis in our model causes no significant alterations of PBF after 5 days. PMID- 8412091 TI - Microvascular changes in experimental gastric stress ulceration: the influence of allopurinol, cimetidine, and misoprostol. AB - The influence of stress on the structure of the gastric mucosal capillary network was investigated in an experimental model using scanning electron microscopy (SEM) to study corrosion casts of the gastric microvasculature. An index of microcirculatory patency was devised to improve the objectivity of SEM. Stress (mean score 42.4 +/- 4.7) severely disrupted the structure of the mucosal capillary network when compared with controls (mean 5.0 +/- 2.9) (P < 0.01). Disruption of the microvascular network was significantly reduced by pretreatment with allopurinol, cimetidine, and misoprostol, these groups having mean damage scores of 17.4 +/- 5.8, 21.2 +/- 7.9, and 28.6 +/- 9.3, respectively, when compared with untreated stressed controls in which the mean score was 42.4 +/- 4.7 (P < 0.02). Microvascular disruption is a significant factor in stress ulceration and the efficacy of allopurinol in minimizing the stress-induced disruption of the microvascular network provides further indirect evidence for the role of ischemia and oxygen-derived free radical generation in its etiology. PMID- 8412092 TI - The role of Kupffer cells in the surveillance of tumor growth in the liver. AB - The present study was designed to investigate the role of Kupffer cells (Kc) in the surveillance of liver tumors. We examined the antitumor activity of Kc by 51Cr releasing assay and inhibition of [3H]thymidine ([3H]TdR) incorporation into tumor cells. We also studied the change in the growth of liver tumors following the activation and the blockade of Kc. The cytotoxicity of Kc against K562 increased as the effector:target (E:T) ratio rose and reached its maximum level of about 18% at an E:T ratio of 20:1. [3H]TdR incorporation into target cells (P815 and AH130) was also inhibited by Kc. Such antitumor activity of Kc was augmented by OK432 (K562, from 13.8 +/- 5.6 to 21.9 +/- 2.5%; AH130, from 19.2 +/ 14.5 to 37.1 +/- 12.6%). In the experiment of the inoculation of AH130 via the portal vein, OK432 decreased the number of hepatic foci, whereas macrophage inhibitors carrageenan and gadolinium increased the number of tumor nodules. In addition, gadolinium injection reduced the number of Kupffer cells reactive with monoclonal antibodies directed against macrophages ED2 and Ki-M2R. Tumor growth in the liver was maximum in rats with both gadolinium treatment and splenectomy. In conclusion, Kc have antitumor activity, and augmentation of Kc may be a possible strategy to prevent hematogenous hepatic metastasis. PMID- 8412093 TI - Lipopolysaccharide detoxification by endotoxin neutralizing protein. AB - Endotoxin neutralizing protein (ENP), a recombinant form of the anti lipopolysaccharide factor that was isolated from amebocytes of the American horseshoe crab, Limulus polyphemus, detoxifies lipopolysaccharide (LPS) both in vitro and in vivo. Using the Limulus amebocyte lysate assay, LPS was detoxified by ENP at a 1 to 1 weight ratio (1:1). When isolated rat aortic rings were preincubated for 16 hr with either LPS or LPS/ENP (1:5), only aortas in the LPS/ENP group contracted normally under norepinephrine stimulation. To show that detoxification of a lethal amount of LPS (18 mg/kg, LD50 at 48 hr) persists in vivo, LPS/albumin (1:1) or LPS/ENP (1:1) mixtures were preincubated (30 min, 37 degrees C) and then injected intravenously into rats. In the 8 hr after injection, LPS/ENP challenged rats, in contrast to their LPS/albumin injected counterparts, had significantly fewer physical signs of acute LPS toxicity (P < 0.001). At 48 hr after challenge, all LPS/ENP treated rats survived (P < 0.01 vs LPS/albumin), and with significantly less weight loss (P < 0.001 vs LPS/albumin challenged survivors). At necropsy, the LPS/ENP group was free of typical LPS induced gross organ lesions, notably in the liver, spleen, gut-associated lymphoid tissue (GALT), and small intestine. By microscopic examination, lymphocytic necrosis in the spleen and GALT of the LPS/ENP treated survivors was significantly milder than that in the LPS/albumin challenged survivors, although the degree of hepatocellular necrosis and small intestinal enteritis was similar. LPS-neutralizing proteins such as ENP may be useful in treating LPS toxicity. PMID- 8412094 TI - Stress distributions in vascular aneurysms: factors affecting risk of aneurysm rupture. AB - Aneurysm rupture occurs when local wall stresses exceed the tensile strength of vascular tissues. Knowledge of vascular wall stresses, and insight into the factors that change wall stresses, will lead to a better understanding of how aneurysms grow and rupture. In this study, stress distributions in the walls of small aneurysms were calculated using finite element analysis (FEA), a numerical technique able to predict stress distributions with greater accuracy than the Law of Laplace. Stresses were calculated for an initial small aneurysm and compared to stresses produced by increasing the aneurysm diameter, decreasing the wall thickness, and changing the material properties of the aneurysm wall. FEA calculations indicate that wall stresses are generally greatest on the inner surface of an aneurysm, and decrease nonlinearly as the outer wall is approached. Maximum wall stresses occur along the region of greatest diameter, and circumferential stresses tend to be significantly greater than longitudinal stresses. Doubling the diameter of an aneurysm produced a twofold increase in the maximum wall stress. Decreasing the wall thickness by half also produced a doubling of the maximum wall stress. Changing material properties produced no appreciable change in wall stresses. However, weaker materials fail at lower stresses, thus halving material strength would be equivalent to doubling wall stresses. We conclude that the Law of Laplace is inaccurate in predicting the complicated stress distributions that exist in aneurysm walls, and that more sophisticated tools, such as FEA, will be needed to understand this complex phenomenon. We also conclude that proportional changes in the diameter, wall thickness, or aneurysm tissue strength have roughly equivalent effects on aneurysm growth and rupture. PMID- 8412095 TI - Pulmonary hemodynamic consequences of ECG-synchronized ventilation. AB - The pulmonary hemodynamic consequences of ECG-synchronized jet ventilation were studied in an acute closed chest swine model (n = 11). Eight jet timing protocols were compared to conventional mechanical ventilation. Hearts were paced atrially at 120 beats per minute, and analog measurements of pulmonary arterial flow and pulmonary arterial, tracheal, pleural, left atrial, and femoral arterial pressure were digitized in real time at 200 Hz. Fourier analysis of pulmonary artery pressure and flow waveforms was employed to calculate mean and oscillatory right ventricular hydraulic power and pulmonary vascular input impedance. Measurements were taken at 0, 5, and 10 cm H2O of positive end-expiratory pressure (PEEP) during conventional respiration and synchronized ventilation modes. No difference was found in mean pulmonary pressure and flow between conventional and synchronized ventilation at any level of PEEP, regardless of the timing of the jet pulse relative to the cardiac cycle. A significant difference in mean tracheal pressure between conventional and jet ventilation could be found only in the absence of PEEP (3.8 +/- 0.5 vs 2.5 +/- 0.3 mm Hg, P < 0.05). In the absence of PEEP, total hydraulic power was significantly less with respect to conventional ventilation when the jet pulse trailed the QRS complex by 90 and 135 degrees. A significant decrease in the ratio of oscillatory-to-mean power versus conventional respiration was found when jet ventilation lagged the QRS by 135 degrees (0.115 +/- 0.015 vs 0.147 +/- 0.013). These differences did not persist when PEEP was added. Moreover, no significant difference in hemodynamic variables was found when the various jet timing protocols were compared to each other.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412096 TI - Endothelins mediate intestinal hypoperfusion during bacteremia. AB - We have previously reported that Escherichia coli bacteremia induces hypoperfusion and vasoconstriction of the rat small intestinal microcirculation. However, the mechanisms which mediate these responses are not clearly defined. Because serum levels of endothelins, a family of potent vasoconstrictor peptides, are increased during bacteremia, we postulated that endothelins contribute to intestinal hypoperfusion during infection. Using intravital microscopy, we characterized the effects of topically applied recombinant endothelin-1 on small intestinal arteriolar diameters and blood flow. Dose-dependent vasoconstriction of both large (A1) and small (A3) arterioles and hypoperfusion were observed. To assess whether endothelins contribute to alterations of the intestinal microcirculation during bacteremia, antiserum was used to inhibit endothelins during E. coli bacteremia. Endothelin inhibition resulted in restoration of blood flow and attenuation of vasoconstriction. Our results suggest that endothelins contribute to intestinal hypoperfusion and arteriolar vasoconstriction during bacteremia. PMID- 8412097 TI - Intestinal migrating myoelectric complexes in rats with acute pancreatitis and bile duct ligation. AB - Intestinal migrating myoelectric complexes (MMC) were studied in rats subjected to common bile duct ligation. Four pairs of bipolar electrodes were implanted on the jejunum and the ileum of 21 rats. Animals were allowed to recover 7 days before electrical recordings were made. Bile duct ligation at the entrance to the duodenum (low) or above the pancreas (high) or sham operation was performed. MMCs were recorded before and 48 and 72 hr after the operation in each animal. MMC intervals were unchanged following high ligation or sham operation. However, 48 hr after low bile duct ligation, the MMC interval (20.6 +/- 2.5 min) was significantly prolonged compared with that before ligation (14.0 +/- 1.2 min). Further prolonged intervals (28.4 +/- 4.2 min) were observed 72 hr after the ligation. There were no significant differences among the slow wave frequencies in any group of animals, being in the range of 34.1 +/- 1.1 to 36.3 +/- 1.0 cycles/min. Thus, the MMC pattern in this rat model may be affected by the development of acute pancreatitis. The mechanism(s) responsible for altering the MMC pattern are not known; however, hepatic biliary secretion alone does not appear to play a primary role. PMID- 8412098 TI - Stress ulceration and gastric mucosal cell kinetics: the influence of prophylaxis against acute stress ulceration. AB - Acute gastric stress ulceration occurs frequently in severely ill patients. Mucosal ischemia is central to the etiology of stress ulceration. The ability of the mucosa to restore continuity when damaged is one of the most important of the local defence mechanisms against injury. The aims of this study were to investigate this restitutive process during stress ulcer formation and to determine the effect of prophylaxis against stress ulceration on gastric mucosal cell kinetics. Nuclear DNA was radiolabeled in vivo with tritiated thymidine, and after autoradiography the position and number of labeled cells along the gastric mucosal gland were determined. Wistar rats were studied immediately and 48 hr after cold restraint stress. The labeling index in the proliferative zone of the gastric mucosa was significantly suppressed immediately after stress (12.58% vs 16.81% for unstressed controls, P < 0.001). This effect persisted for 48 hr after stress (9.89% vs 16.35% for unstressed controls, P < 0.001). Prophylaxis against stress ulceration with cimetidine or allopurinol prevented this suppression of gastric mucosal cell kinetics and promoted early migration of labeled cells towards the surface of the mucosal gland. Allopurinol prophylaxis was associated with migration of mucosal cells immediately following stress greater than that following cimetidine prophylaxis (14.0% vs 9.3% surface layer labeling index, P < 0.01). Allopurinol, a xanthine oxidase inhibitor, reduces oxygen free radical production during ischemia reperfusion injury. These results emphasise the importance of the gastric mucosal defence mechanisms in protection against injury and indicate the role of ischemia in the aetiology of acute gastric stress ulceration. PMID- 8412099 TI - Effect of age upon ischemia/reperfusion injury in rat muscle free flaps. AB - Metabolic and functional changes have been found in tissues of aging man and animals. It is not known if old age has a detrimental effect on the outcome of free tissue transfer or extremity replantation, nor has it been determined if prolonged ischemia may exacerbate such effects. To explore these issues, we utilized a syngeneic rat model of cutaneous maximus muscle transplantation, isolating the effects of flap age and ischemia on flap survival and metabolic function. Flaps were raised in young adult (2-3 months), middle-aged (10-12 months), and old (20-22 months) Lewis rats and transplanted to young Lewis recipients after 1, 6, or 10 hr of room temperature ischemia. Reperfusion periods of 2 hr, 2 days, or 2 weeks were allowed, and flaps were harvested for histologic and histochemical evaluation. Flap weights significantly increased after reperfusion following longer vs shorter ischemia and in the old flaps versus young flaps (for 1 hr of ischemia) (P < 0.05). Histology confirmed a greater extent of interstitial edema in flaps from older rats. Histochemical assessment of muscle dehydrogenase activity (nitroblue tetrazolium staining) demonstrated reduced staining in both the young 10-hr ischemic flaps and in the older 6- and 10-hr ischemic flaps. These results indicate that muscle does not tolerate ischemia as well in older animals, but that a short ischemic interval (1 hr) is well-tolerated. PMID- 8412100 TI - Effect of diltiazem on altered glucose regulation during endotoxic shock. AB - Endotoxic shock is associated with profound metabolic alterations including hypoglycemia and hyperlactiacidemia. We studied hepatic glucose production and the role of diltiazem in affecting these metabolic alterations in a rat model of endotoxic shock. Fasted rats were intravenously injected with saline, endotoxin (20 mg/kg), endotoxin (20 mg/kg) plus diltiazem (1.2 mg/kg), or saline plus diltiazem. Five hours after the injections, the rats were anesthetized and blood samples were obtained for plasma glucose and lactate. The livers of all rats were then perfused in situ with an oxygenated 37 degrees C glucose-free Hanks' balanced salt solution at a rate of 30 ml/min via the portal vein and effluent was collected from the inferior vena cava. After 30 min equilibration, 5 mM lactate was added to the perfusate as a gluconeogenic substrate. Endotoxic groups exhibited hypoglycemia (64.5 +/- 12.7 mg/dl) and hyperlacticacidemia (4.3 +/- 0.63 mM). Diltiazem administration improved the hypoglycemia (96.9 +/- 9.1 mg/dl) and attenuated the hyperlacticacidemia (2.2 +/- 0.7 mM) in the endotoxic group. Gluconeogenic stimulation with lactate was demonstrated in both the control (18.1 +/- 2.3 vs 12.8 +/- 1.86 microns/g/hr, P < .05) and endotoxic (16.6 +/- 2.3 vs 9.8 +/- 1.1, P < .05) groups. However, stimulation in the endotoxic groups was significantly less compared with control. Gluconeogenic stimulation in the endotoxic group was unaffected after diltiazem administration. These data suggest that diltiazem treatment in endotoxic shock improves hypoglycemia and attenuates hyperlacticacidemia. These metabolic alterations are not associated with an improvement in substrate-specific stimulation of gluconeogenesis. PMID- 8412101 TI - Evaluation of NR-LU-10 with the Neoprobe gamma detector in a mouse model. AB - The Neoprobe Model 1000 hand-held gamma detector, in combination with the murine monoclonal antibody (MAb) B72.3 can successfully intraoperatively target both primary and recurrent colorectal cancer. Because of the shortcomings of this system (length of time needed to clear unbound MAb and heterogeneous staining of cancer cells), new MAbs are under investigation. NR-LU-10 IgG and its FAB fragment were evaluated with the Neoprobe gamma detector in a nude mouse model. NR-LU-10 is a pancarcinoma IgG2b antibody that recognizes an oncofetal glycoprotein antigen that is expressed by most carcinomas. Animals were injected intraperitoneally with 125I-labeled IgG, 125I, or technetium 99m (99mTc)-labeled Fab fragment. The Neoprobe gamma detector and standard gamma well counts provided biodistribution and pharmacokinetic data. The Fab fragment achieved a 5:1 tumor to blood-pool background (BPB) ratio in only 66 hr, whereas the whole MAb required 14 days. Technetium did not appear to be an adequate isotope for this system because of its short half-life. Autoradiographs performed for both the 125I-labeled NR-LU-10 IgG and its 125I-labeled Fab fragment (but not 99mTc labeled Fab) localized well in this model and gave adequate tumor to BPB and tissue ratios. Use of the 125I-labeled Fab significantly decreased the time required for clearance of the MAb radionuclide complex from the blood and tissue background, thus providing earlier tumor localization in the Radioimmunoguided Surgery (RIGS) system. PMID- 8412102 TI - Regulation of collagen gene expression in keloids and hypertrophic scars. AB - The synthesis of type I and III collagens in cultured skin fibroblasts from normal skin, normal scar, hypertrophic scar, and keloids was examined. The ratio of type I/III collagen was significantly elevated in keloids compared to that in the other groups. When mRNA steady-state levels coding for alpha 1(I) procollagen were determined, it was apparent that this increase in the type I/III collagen ratio in keloids was paralleled by a specific increase in alpha 1(I) procollagen mRNA. This specific increase in alpha 1(I) procollagen mRNA in keloids was the result of increased gene expression because the transcription rate of the alpha 1(I) procollagen gene was significantly elevated in keloids, as determined by nuclear runoff transcription. The rate of transcription of the alpha 1(I) procollagen gene was also elevated in hypertrophic scars, although no concomitant increase in alpha 1(I) procollagen mRNA levels or alteration in the type I/III collagen ratio was observed. These data indicate that the rate of gene transcription of alpha 1(I) procollagen is increased in both hypertrophic scars and keloids, but only keloids exhibit increased steady-state levels of alpha 1(I) procollagen mRNA and concurrent increases in type I collagen. These results suggest that at least two distinct mechanisms, one pretranscriptional and one post-transcriptional, regulate type I collagen synthesis. It is possible, therefore, that in keloids, neither mechanism functions efficiently to down regulate type I collagen. In hypertrophic scars, however, the post transcriptional mechanisms are able to decrease elevated levels of mRNA coding for alpha 1(I) procollagen that result from increased transcription of the alpha 1(I) procollagen gene. PMID- 8412103 TI - Regulation of gastric H(+)-K(+)-ATPase by cAMP-dependent protein kinase. AB - A functional approach was utilized to isolate protein effectors from cAMP stimulated rabbit gastric microsomes capable of stimulating H(+)-K(+)-ATPase activity. These studies have resulted in isolation of a cAMP-dependent protein kinase product from rabbit gastric microsomes which is capable of stimulating the proton pump of the parietal cell, H(+)-K(+)-ATPase, in inhibited gastric microsomes. This protein is membrane-bound and may be extracted from gastric microsomes only in the phosphorylated state. This phosphoprotein has at least 20 phosphorylation sites and produces enhancement of H(+)-K(+)-ATPase activity which equals that induced by the K+ ionophore, valinomycin. It would appear, therefore, that cAMP-mediated acid secretion involves phosphorylation of a membrane-bound cAMP-dependent protein kinase substrate in close proximity to the proton pump which produces K+ conductance and thereby controls the rate of acid secretion. The degree of phosphorylation of this protein is probably controlled by the activities of cAMP-dependent protein kinase and phosphoprotein phosphatase. PMID- 8412104 TI - Effectiveness and tolerance to highly concentrated vs conventional TPN formulas. AB - Conventional total parenteral nutrition regimens (TPN-C) involve concentrations of dextrose/protein which necessitate administration of 2.5-4 liters/day to meet target nutritional needs. Although this is frequently acceptable, certain clinical settings mandate a volume-restricted (VR) approach. This study compares a VR TPN regimen (TPN-VR) involving the use of 25% dextrose and 9.5% amino acid with D17.5 AA 5.0 (TPN-C). The two groups were compared for adequacy of nutritional delivery, balance, and tolerance. Twenty patients received the TPN-VR (Group 1) and 20 patients received TPN-C (Group 2). The groups were comparable in age, sex, injury severity, and APACHE 2 scores. Harris-Benedict (BEE) x 2 and 2 g/protein/kg of ideal body weight were delivered by the second day of TPN. A 27% reduction in administered fluid was achieved in Group 1 (P < 0.001). Metabolic cart data in both groups demonstrated that delivered calories exceeded REE. The average RQ in Group 1 was 0.84 and in Group 2 was 0.90 (P > 0.1). There was no significant difference between the two groups in nitrogen balance, mean serum bilirubin levels, PT and PTT, serum albumin levels, and triglycerides (P > 0.20). SGPT and alkaline phosphatase levels were significantly higher in Group 2 (P < 0.001). Group 2 received an average of 22% more carbohydrate than Group 1 and 45% required insulin compared to 25% in Group 1 (P < 0.01). In summary, TPN-VR is comparable to TPN-C in terms of effectiveness of delivery, nutritional balance, and tolerance. PMID- 8412105 TI - Mechanistic model of wound contraction. AB - The interplay between cellular, biochemical, and biomechanical phenomena which result in wound contraction is examined with the aid of a mathematical model. The model accounts for fibroblasts repopulating the wound extracellular matrix (ECM) by a combination of migration and proliferation and, in the course of exerting traction forces associated with their motility, their deformation of the local ECM, which has a viscoelastic character. A base model, which assumes that cell and tissue properties are the same irrespective of position with respect to the wound center, is found to be inconsistent with wound contraction. Extensions of the base model which account for the possible influence of a concentration gradient of an inflammation-derived mediator on the traction, growth, or chemotactic properties of the fibroblasts do predict the qualitative features of a contracting wound. In particular, the case where traction is prescribed to increase as fibroblasts approach the wound center is shown to obey the commonly observed exponential rate contraction law and to be largely consistent with the findings of McGrath and Simon (Plast. Reconstr. Surg. 72, 66, 1983) concerning the independence of the contraction rate constant and final extent of contraction from initial wound size and geometry. PMID- 8412106 TI - Expression of colorectal carcinoma-associated antigens in colonic polyps. AB - Immunohistologic techniques were used to study the expression of colorectal carcinoma-associated antigens in colonic polyps and to compare this with expression in the normal colonic epithelium. Forty-nine polyps were studied using monoclonal antibodies to 16 different blood group and differentiation antigens and carcinoembryonic antigen epitopes. With the Lewis(a) antigen and the two epitopes of CEA recognized by 3D6 and COL-4 expression in polyp tissue was the same as that in the normal colon. Five types of alteration of antigen expression in polyps were seen. The blood group antigens A, B, and Lewis(b), which are expressed only on the right side of the normal adult colon, were detected in both neoplastic and nonneoplastic polyps from the distal colon. The Lewis(x) antigen and the antigen epitopes detected by the antibodies COL-12, CA19-9, ME491, and GA73.3 showed an increased frequency of expression in all types of polyps in comparison with the normal colonic epithelium, while H-type 2, ND4, and the antigen epitope detected by CO29.11 showed a slightly decreased frequency of expression in polyp tissue. The X-like antigen which was expressed in only 7% of normal colon specimens showed increased frequency of expression in polyp tissue with significantly greater expression in neoplastic than hyperplastic lesions (P = 0.003). The TAG-72 antigen was detected only in adenomas with severe dysplasia (P = 0.01), correlating well with premalignant histology. These findings have helped us clarify the variation of antigen expression in colonic polyps and allowed us to define which antigens are worthy of further investigation as markers of possible malignant transformation. PMID- 8412107 TI - Transient protection of incomplete colonic anastomoses with fibrin sealant: an experimental study in the rat. AB - Fibrin glue has been used as a protective seal in normal and high-risk anastomoses to prevent leakage. The influence of fibrin adhesive on the healing colonic anastomosis in a control and high-risk model was tested. Resection and anastomosis of the left colon was performed in rats. In group Ia an end-to-end anastomosis was constructed with 12 7-O polypropylene sutures; in group Ib the anastomosis was sealed with fibrin adhesive. In group II an incomplete anastomosis was constructed with only 4 sutures at 90 degrees, therefore potentially leaking. In group IIb additional sealing with fibrin glue was performed. On Days 2, 4, and 7 body weight, adhesion formation, anastomotic bursting pressure, and collagen concentration were measured. The results showed increased adhesion formation after fibrin sealing. The anastomotic bursting pressure of incomplete anastomoses showed a significant increase after sealing on Day 2 only; on Day 4 and 7 no differences were found. Sealing of control anastomoses caused lower bursting pressures on Day 4. Collagen concentration is significantly reduced after fibrin sealing of normal anastomoses. We conclude that fibrin sealing of control anastomoses inhibits wound healing. Incomplete anastomoses are temporarily protected by fibrin glue sealing. Finally, fibrin sealing of the colon wound does not prevent adhesion formation. PMID- 8412108 TI - Chronic endotoxemia reversibly alters respiratory burst activity of circulating neutrophils. AB - Endotoxin-induced sequestration of neutrophils and ectopic activation of the cytocidal respiratory burst are thought to contribute to the pathophysiology of "endotoxin shock." The effects of endotoxin on the circulating neutrophil population have not been described. A rat model of continuous sublethal endotoxin infusion by minipump was used to study spontaneous and stimulus-specific activation and priming of superoxide anion and chemiluminescence production of circulating neutrophils. At 3 hr of endotoxin infusion there are marked neutropenia, minimal spontaneous activation compared to an active response in the control, and no priming effect. By 30 hr of endotoxin infusion, a neutrophilic leukocytosis, a marked spontaneous activation of superoxide anion and chemiluminescence production are observed, with no priming response and a loss of zymosan-stimulated chemiluminescence. The biologic significance of endotoxin induced spontaneous activation is unknown. By 78 hr of infusion the host tolerance to endotoxemia is also evident for neutrophils, since neutrophil responses are nearly identical in endotoxin- and saline-infused groups. The loss of zymosan-stimulated chemiluminescence in both groups at 78 hr suggests a foreign-body effect from the minipump and may reflect altered arachidonic acid metabolism. PMID- 8412109 TI - Effect of blood transfusion on immune function. IX. Effect on lymphocyte metabolism. AB - Blood transfusions have been repeatedly shown to be immunosuppressive in nature. The intracellular mechanisms of this immunosuppression have not been extensively investigated. We investigated the effect of blood transfusions on lymphocyte intracellular metabolism of glucose and amino acids, as well as levels of adenosine deaminase activity and nucleotide triphosphate concentrations. Blood transfusions were found to increase the rate of glucose and glutamine metabolism, to increase nucleotide triphosphate concentrations, and to increase the level of adenosine deaminase activity. This increased level of lymphocyte metabolism in the face of immunosuppression would appear to indicate that the transfusion induced immunosuppression is an active dynamic process. PMID- 8412110 TI - Effect on liver and splanchnic circulation of graded flow rates through portal systemic bypass during acute portal occlusion. AB - Using graded flow rates through a passive portal-systemic bypass during 2-hr portal occlusion in dogs, splanchnic, hemodynamic, and metabolic changes and liver insults were evaluated to determine critical bypass flow. To exclude effects caused by reduced circulating blood volume, constant preocclusion levels of cardiac output were maintained throughout the study by increasing intravenous infusion rates. With the decrease in bypass flow rates during portal occlusion, portal pressure increased, superior mesenteric arterial flow decreased, and hepatic arterial flow rapidly increased and these parameters maintained their individual levels during occlusion. These hemodynamic parameters recovered to nearly normal levels 3 hr after release of the portal clamp. Metabolic acidosis progressed with decreasing bypass flow, but portal potassium and inorganic phosphorus levels showed a significant rise only when there was no bypass. Portal levels of creatine phosphokinase BB, beta-glucuronidase, and endotoxin did not show significant changes corresponding to bypass flow. The amounts of infusion required were 3 to 6 times the basal level (10 ml/kg/hr) during occlusion and 1.7 to 3 times after release in 30% or less bypass groups. Upon ceasing infusion 3 hr after release, dogs in the 10% or less bypass groups underwent circulatory insufficiency. Changes in total adenine nucleotide and energy charge ratio of the liver, postoperative changes in transaminase levels, and animal survival indicated that a 2-hr interruption of the portal flow, either sufficiently or insufficiently bypassed, caused only minimal insults of the liver.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412111 TI - Free radical scavengers prevent intestinal ischemia-reperfusion-mediated cardiac dysfunction. AB - Our previous studies show that intestinal ischemia impairs cardiac function. This present study examined the contribution of oxygen-derived free radicals to cardiac dysfunction after intestinal ischemia-reperfusion in a rat model of superior mesenteric artery (SMA) occlusion (atraumatic clip for 20 min) and ligation of collateral arcades from the right colic and jejunal arteries. Controls were sham operated (Group 1, n = 10); in Group 2, 20 rats with SMA occlusion were sacrificed 2-5 hr after reperfusion without treatment. Superoxide dismutase (SOD) and catalase, scavengers of oxygen-derived free radicals which have been shown to effectively reduce ischemic injury in several models of traumatic injury, were given as 6000 units/350 g body wt either 1 min after SMA occlusion (Group 3, n = 11) or 2 min after reperfusion (Group 4, n = 10). To examine the contribution of neutrophils as a source of free radicals, additional groups of animals were treated with pentoxifylline (PTX, a methylxanthine derivative which has been shown to decrease neutrophil adherence and aggregation as well as to decrease superoxide production) either 1 min after SMA occlusion (Group 5, n = 10) or 2 min after reperfusion (Group 6, n = 10). Cardiac contractile depression occurred in the untreated ischemic group as indicated by a fall in left ventricular pressure (from 77 +/- 3 to 63 +/- 4 mm Hg, P < 0.01) and +dP/dt max (from 1827 +/- 60 to 1558 +/- 98 mm Hg/sec, P < 0.03) and -dP/dt max (from 1267 +/- 57 to 953 +/- 68 mm Hg/sec, P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412112 TI - Effect of mixing ratio in lecithin/bile acid mixed micelles on Na(+)-K+ ATPase. AB - Lecithin and bile acid form mixed micelles in the bile. We found that the mixing ratio in lecithin/bile acid mixed micelles directly changes the activity of Na(+) K+ ATPase. Na(+)-K+ ATPase activity was suppressed to 13.3% of that in the buffer alone at a lecithin/bile acid mixing ratio of 0.1 in the presence of 10(-2) M bile acid (that is, a concentration level close to that of the hepatic bile). With increase in the mixing ratio in the presence of 10(-2) M bile acid, the activity of the enzyme was found to be augmented accordingly. Thus addition of lecithin to an extent such that the mixing ratio reached 0.6 led to an enhancement of enzymatic activity to 193.8%. If lecithin addition is made in the presence of bile acid at the near gallbladder concentration of 10(-1) M, however, the Na(+)-K+ ATPase activity can be observed to increase with the increase in the lecithin/bile acid ratio. Yet, at the same mixing ratio of 0.6, the enzyme activity was arrested at only 25.7%. However, this change in mixing ratio had no effect on ouabain-insensitive ATPase. The state of mixed micelles may exert the same effect on Na(+)-K+ ATPase activity in vivo. PMID- 8412113 TI - Endothelin does not contribute to the vasospasm after balloon angioplasty in vitro. AB - The present study was undertaken in 43 rabbits to determine whether endothelin or potassium are mediating substances for vasospasm originating from the vessel wall after balloon angioplasty. Thirteen aortic segments were dilated and the luminal surfaces perfused. These perfusates were investigated for their vasomotor action per se and analyzed for their concentrations of potassium and endothelin-like immunoreactivity (ET-LI). The perfusates from angioplasty segments caused a contraction of 2.4 +/- 0.8 mN, while the perfusates from control segments only resulted in a contraction of 0.4 +/- 0.2 mN, (P = 0.004). There was no difference in the concentrations of potassium or ET-LI in the perfusates. Thirty aortic segments were used to measure the concentration of ET-LI in extractions of untreated, control, and angioplasty specimens. There was no statistically significant difference between the groups. Scanning electron microscopy demonstrated the angioplasty segments to be denuded of their endothelium. In conclusion, perfusates from rabbit aortic segments treated by angioplasty in vitro have a strong vasoconstricting effect, for which neither endothelin nor potassium seems to be responsible. PMID- 8412114 TI - Calcium effects on human esophageal smooth and striated muscle. AB - Because of conflicting results in earlier investigations, our study was carried out to determine the effects of acute hypercalcemia on esophageal muscle function at the lower esophageal sphincter and on the smooth and striated muscle of the esophageal body. Variables measured included lower esophageal sphincter pressure and length and esophageal body contraction pressure, duration, and velocity in the middle, distal, and proximal esophagus. Despite 3 hr of induced acute hypercalcemia, there were no statistically significant changes in any of the variables. Although esophageal muscle contraction requires calcium, our results, in contrast to some earlier studies, suggest that the availability of excess calcium does not effect esophageal function. PMID- 8412115 TI - An eosinophil chemotactic lymphokine in the supernatant of lymphokine-activated killer (LAK) cell culture: relationship to interleukin-5. AB - The administration of interleukin-2 (IL-2) systemically or locally to cancer patients, either alone or in combination with lymphokine-activated killer (LAK) cells, frequently results in eosinophilia. To investigate the mechanism of such eosinophil accumulation, we measured in vitro eosinophil chemotactic activity (ECA) in the supernatant of LAK cell cultures by a modified Boyden's chamber method. Consequently, a potent and specific ECA was found in the cell-free supernatant of LAK cell cultures. On the other hand, no ECA was found in the supernatant of normal lymphocyte cultures, nor in IL-2 itself. This activity plateaued when 3 x 10(6) lymphocytes were incubated with 1000 U/ml IL-2 for more than 3 days. Sephadex G-100 gel chromatography indicated that the molecular weight of this eosinophil chemotactic factor (ECF) was approximately 45,000 Da. The ECF was stable when heated to 56 degrees C for 30 min but was inactivated by trypsin, indicating that the ECF is a lymphokine which has very similar characteristics to those of IL-5. Using ELISA for hIL-5, the IL-5 level in the supernatant of LAK cell cultures increased dose dependently with increasing IL-2 concentrations. These results suggest that IL-5 may be responsible for the local recruitment of eosinophils in IL-2/LAK therapy. PMID- 8412116 TI - Spontaneous obesity and increased bile saturation in the ground squirrel. AB - Obesity is a well-known risk factor for developing cholesterol gallstones, but only indirect human studies are available. Presently available animal models of cholesterol gallstone disease do not become obese under normal conditions, while genetically obese or dietarily manipulated rats lack a gallbladder. The Richardson ground squirrel on a high-cholesterol diet develops impaired gallbladder contractility, excess cholesterol in its bile, and then cholesterol gallstones. To determine if ground squirrels would spontaneously become obese and serve as an appropriate model, animal weights were followed over several months while being maintained under a regulated laboratory environment. The five obese animals were twice the body weight of the five lean controls and also had a 20% rise in body fat. Plasma cholesterol levels were markedly increased and liver histology revealed mild microvesicular steatosis. Cholesterol saturation rose in the gallbladder bile of the obese group. This alteration was primarily due to a significant increase in phospholipid (33%) and cholesterol (86%) levels. Such a laboratory environment interrupts the natural annual rhythm of the ground squirrel and prevents hibernation. Animals continued to eat and develop severe obesity and abnormal gallbladder bile. These spontaneously obese animals provide an excellent means for examining altered hepatobiliary secretion and gallbladder motility relative to cholesterol gallstone disease. PMID- 8412117 TI - Gastric stasis of solids after Roux gastrectomy: is the jejunal transection important? AB - To investigate the effect of jejunal transection on the rate of gastric emptying after Roux gastrectomy, a two-part study was conducted. First, we investigated the relationship between Roux limb slow wave frequency and gastric emptying of solids. Six dogs underwent Roux-en-Y gastrectomy with vagal preservation and placement of intestinal electrodes. Gastric emptying studies were performed on each animal with simultaneous pacing of the Roux limb, either at the slowest rate (Pmin) or the fastest rate (Pmax) at which entrainment could be achieved. Gastric emptying studies were also performed in the unpaced (control) condition. Gastric half-emptying times (X +/- SEM minutes) and slow wave frequencies (X +/- SEM cycles per minute), respectively, were Pmin 117 +/- 26 min, 15.7 +/- 0.1 cpm; Pmax 97 +/- 18 min, 19.0 +/- 0.3 cpm; and unpaced 127 +/- 16 min, 15.1 +/- 0.3 cpm. The gastric half-emptying time during Pmax was significantly lower than unpaced controls (P = 0.01). The second part of the study sought to determine if transecting the intestine at 10 cm distal to the pylorus rather than at 20 cm distal to the ligament of Treitz would improve gastric emptying in animals with a truncal vagotomy and Roux-en-Y gastrectomy. Gastric half-emptying times were 149 +/- 21 and 164 +/- 24 min (ns), respectively. Slow wave frequencies were 17.01 +/ 0.06 and 15.7 +/- 0.17 cpm (P < 0.05), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412118 TI - A 5-HT2 receptor mediates serotonin-induced electrolyte transport in rat left colon. AB - Serotonin (5-hydroxytryptamine, 5-HT) is a potent mediator of diarrhea in the carcinoid syndrome. However, the mechanisms of serotonin-induced intestinal secretion remain unclear. The purpose of this study was to determine whether 5-HT stimulates electrolyte transport by a direct action at a mucosal receptor. Serotonin-stimulated electrolyte transport was studied in flat sheet preparations of rat left colon using flux chambers. 5-HT stimulated a concentration-dependent rise in short-circuit current in sheets stripped of muscularis propria. The half maximal concentration required to produce this effect was 50 microM. The preferential 5-HT2 antagonist ketanserin produced a rightward shift of the serotonin concentration-response curve with a pA2 value of 7.7. Serotonin stimulated electrolyte transport by inhibiting mucosa to serosa movement of both sodium and chloride. The 5-HT1 antagonist N-acetyl-5-hydroxytryptophyl 5 hydroxytryptophan amide and the 5-HT3 antagonist 3-tropanyl-indole-3-carboxylate methiodide had no effect on these actions of serotonin on electrolyte transport. In contrast, ketanserin inhibited the actions of serotonin on both sodium and chloride movement and inhibited the rise in short-circuit current induced by 100 microM 5-HT. This study demonstrates that a 5-HT2 receptor located in the vicinity of the mucosa is involved in the regulation of serotonin-stimulated intestinal electrolyte transport. PMID- 8412119 TI - Extremity metabolism in the cachectic, VX-2 carcinoma-bearing rabbit. AB - The pathophysiology of skeletal muscle loss in cancer cachexia is poorly understood. Immature, male, New Zealand White rabbits (TBs; n = 11) were implanted with VX-2 carcinoma and various indices of systemic and limb metabolism were examined in comparison with pair-fed controls (PFCs; n = 9) and normal controls (NCs; n = 22) fed ad lib. The TBs became hypophagic and experienced reduced growth relative to both control groups (P << 0.001). At 7 weeks (tumor burden 3-6% of body weight; no metastasis) the TBs had the following statistically significant differences from NCs: anemia, neutrophilic granulocytosis and thrombocytosis, hypercalcemia, hypoinsulinemia, elevated plasma triglycerides and altered plasma amino acids, increased hind limb effluxes of lactate and most amino acids. These alterations were not caused by hypophagia, since the PFCs were normal at 7 weeks with regard to all measured parameters except body weight and limb flow, both of which were reduced. The decrease in flow (P < 0.05) apparently contributed to conservation of skeletal muscle amino acids in the PFCs. Young New Zealand White rabbits implanted with VX-2 carcinoma manifest tumor burden, wasting, and metabolic alterations qualitatively similar to those seen with many human cancers. PMID- 8412120 TI - Tissue oxygenation in hypovolemic shock. AB - Bowel mucosal ischemia may be related to the development of systemic sepsis. Traditional indices of oxygen metabolism are flow-weighted averages which do not reflect tissue-specific oxygen concentration. We undertook this study to examine the relationship between systemic oxygen delivery (DO2) and tissue oxygen tension (TPO2) in hypovolemic shock. A modified Wiggers model was used to produce hypotension in five swine. TPO2 was measured continuously with fluorescence quenching 1-mm probes placed in the submucosa of the terminal ileum and subcutaneously in an axillary fold. Shock was maintained for 1 hr, followed by resuscitation. Cardiac output, systemic and pulmonary arterial pressures, and arterial and mixed venous blood gases were measured every 15 min. Data were analyzed by nonparametric ANOVA and rank coefficients, with logarithmic curve fitting and linear regression. DO2 decreased with phlebotomy (P < 0.003) as did skin TPO2 (P < 0.001) and bowel TPO2 (P < 0.0004). Skin and bowel TPO2 varied with DO2 and each other (P < 0.05). TPO2 remained low throughout the shock period and returned to or exceeded baseline levels with resuscitation (P < 0.05). The following were concluded: (1) Hypovolemic shock produces a significant and rapid decrease in subcutaneous and bowel TPO2 with concomitant change in DO2. (2) The degree of bowel ischemia, clinically inaccessible for quantitation, is paralleled by subcutaneous TPO2. (3) TPO2 provides information about oxygen availability in shock and resuscitation not available from traditional parameters of oxygen transport. PMID- 8412121 TI - The reversibility of impaired prostacyclin production of the vein graft. AB - The effects of changes in shear stress (shear stress variation) on production of prostacyclin (PGI2) were examined in canine autologous vein grafts, which were implanted in the poor or normal distal runoff limbs. Four weeks after grafting, the vein grafts were perfused ex vivo and PGI2 was assayed as 6-ketoprostaglandin F1 alpha. The vein grafts were perfused under constant flow for the first 30 min and then under pulsatile flow simulating an abnormal flow with a low shear stress variation or a normal flow with a high shear stress variation for the next 30 min. Basal production rates (30-min cumulative PGI2 production) of vein grafts implanted in the poor runoff and normal runoff limbs were 1.97 +/- 0.71 and 2.19 +/- 0.40 ng/cm2, respectively, with no significant difference. Pulsatile flow effects (increased PGI2 production between 30 and 60 min) of simulated abnormal versus normal flow were 0.50 +/- 0.50 ng/cm2 versus 2.31 +/- 1.87 ng/cm2 in vein grafts implanted in poor runoff limbs (P < 0.05) and 0.48 +/- 0.41 ng/cm2 versus 3.48 +/- 1.08 ng/cm2 in vein grafts implanted in normal runoff limbs (P < 0.01), respectively. There were no significant differences in simulated normal flow effects between poor runoff limbs and normal runoff limbs. The results indicate that the release of PGI2 in vein grafts may decrease in the presence of an abnormal blood flow with a low shear stress variation. In addition, it is suggested that even vein grafts implanted in the poor runoff could increase PGI2 production once the grafts were placed into normal arterial circulation. PMID- 8412122 TI - Immunocytologic analysis of cells obtained from bronchoalveolar lavage in a model of rat lung allograft rejection. AB - Left lung transplantation (n = 30) between BN rat (RT1n) and LEW rat (RT1(1)) was performed to examine serial changes in the inflammatory cell profile and T-cell subsets occurring in bronchoalveolar lavage (BAL) obtained after transplantation. Transplanted animals were sacrificed on Days 1 to 7 post-transplantation. Previous studies show that lung allografts between these rat strains were strongly rejected within 7 days. The serial change in the cell profile of BAL showed a marked initial predominance of polymorphonuclear leukocytes, a decrease in macrophages, and a temporary increase in number of eosinophils on Day 2 post transplantation. A gradual increase in lymphocytes coincident with progression of rejection was also noted. Immunocytologic studies using monoclonal antibodies specific for rat T-cell subsets and interleukin-2 receptor (IL-2R) showed significant increase in pan-T-cells on Days 3, 4, and 5 and T suppressor/cytotoxic (CD8 positive) fraction on Days 4 and 5, whereas the T helper (CD4 positive) fraction peaked on Day 2. The frequency of T-cells expressing IL-2R (55 kDa), indicating activated T-cells, significantly increased as early as Day 2 and maintained its high value thereafter. mRNA levels for IL-2R were detectable in the allografts on Day 2 and peaked on Day 5 post transplantation. The value of CD4/CD8 T-cell ratios rose initially and then dropped below 1.0 on Days 4 and 5. These values differed markedly from those of syngeneic transplants (Lew-->Lew) examined on Day 5 post-transplantation. First, no significant changes in BAL cytology were seen when syngeneic transplants were compared with normal (Lew) lung.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412123 TI - Tumor-induced alterations in hepatic malic enzyme and carnitine palmitoyltransferase activity. AB - To better understand the role of the liver in the hypertriglyceridemia observed in a tumor-bearing state, we have examined tumor-induced alterations in hepatic lipogenesis and fatty acid oxidation. The effects of differing tumor burden as well as tumor excision on the activity and mRNA levels of malic enzyme and carnitine palmitoyltransferase were studied in Fisher 344 rats bearing a methylcholanthrene-induced sarcoma. Serum triacylglycerols and plasma nonesterified fatty acids (NEFA) levels were both elevated with increasing tumor burden (P < 0.05 vs control). The elevation disappeared with tumor removal. Malic enzyme activity of tumor bearers, compared with control rats, declined with an increase in tumor burden. These two variables were negatively correlated (r = 0.90, P < 0.01). The changes in activity were accompanied by positively correlated changes in mRNA levels (r = 0.73, P < 0.01). Carnitine palmitoyltransferase activity was not altered, even in the presence of a large tumor burden. Hepatic lipogenesis, reflected by malic enzyme activity, was tumor dependent and was significantly reduced during the period of circulating hypertriglyceridemia. Fatty acid oxidation, reflected by carnitine palmitoyltransferase activity, was not enhanced in spite of an ample supply of NEFAs to the liver from the peripheral tissues. The data suggest that neither hepatic lipogenesis nor fatty acid oxidation contribute to hypertriglyceridemia in the tumor-bearing state. PMID- 8412124 TI - Rat liver lipids during ex vivo warm and cold ischemia and reperfusion. AB - Rat livers were flushed and stored ex vivo in Krebs-Henseleit buffer at 37 degrees C for 3 hr or in University of Wisconsin solution at 2 degrees C for 48 hr. After this they were perfused with recirculated Krebs-Henseleit solution at 37 degrees C for 1 hr. Levels of phospholipids (PL), free fatty acids (FFA), and conjugated dienes were determined at various times during ischemia and after 1 hr of reperfusion. After 3 hr warm ischemia, total PL content decreased by about 30% primarily because of decreases in phosphatidylcholine and phosphatidylethanolamine. One hour of reperfusion normalized PL levels. Total PL content was unchanged up to 48 hr of cold ischemia because of offsetting alterations in levels of PL classes. FFA accumulation during warm ischemia was about half that during cold ischemia. Conjugated diene concentration increased fivefold during warm ischemia but was unchanged during cold ischemia. Low PL levels and FFA accumulation along with production of conjugated dienes suggest that lipid oxidation is a major mechanism of PL degradation during warm, but not cold, ischemia of the liver. PMID- 8412125 TI - Altered fluidity of liver plasma membranes following partial hepatectomy in rats. AB - The fluidity of liver plasma membranes was assessed during regeneration following partial hepatectomy in rats. Fluorescence polarization was measured using 1,6 diphenyl-1,3,5-hexatriene to evaluate the fluidity of liver plasma membranes. The fluorescence polarization value of liver plasma membranes of control rats was 0.189 +/- 0.005 (mean +/- SEM). The fluorescence polarization values decreased initially after a 70% hepatectomy [0.176 +/- 0.003 at 3 hr and 0.174 +/- 0.004 at 6 hr (P < 0.01)] at 3 and 6 hours, but then recovered to preoperative levels (0.181 +/- 0.004, 0.189 +/- 0.004, 0.189 +/- 0.003, 0.193 +/- 0.003, 0.200 +/- 0.002, 0.208 +/- 0.007, and 0.190 +/- 0.003 at 12, 24, 36, 48, 72, 96 hr, and 7 days, respectively). Flow cytometric analysis using anti-bromodeoxyuridine (BrdU) monoclonal antibody was used to investigate the cell kinetics of the regenerating liver after 70% hepatectomy. The proportion of S-phase incorporating BrdU was not increased at 6 or 12 hr, but it then increased to peak of 27.9 +/- 1.9% at 24 hr. Subsequently, at 48, 72, and 168 hr, the proportion decreased to 8.5 +/- 0.6, 5.9 +/- 0.3, and 1.0 +/- 0.1%, respectively. The ratio of phospholipid to cholesterol contents of liver plasma membranes following partial hepatectomy did not significantly change in spite of markedly altered membrane fluidity. Thus, the alteration of membrane function occurs in conjunction with the initiation of the regeneration process following partial hepatectomy. It is suggested that the alteration of membrane fluidity plays an important role in the homeostatic response for cell proliferation. PMID- 8412126 TI - Modulation of cytotoxic activity of resident macrophages by postsurgical macrophages. AB - The purpose of this study was to determine if the secretion of cytotoxic molecules, such as tumor necrosis factor or toxic oxygen molecules, by resident peritoneal macrophages is modulated by postsurgical macrophages elicited by peritoneal trauma. Resident macrophages were collected from nonsurgical rabbits and cultured in vitro with either spent media from cultures of postsurgical macrophages harvested at various times or with varying concentrations of standard cytokines. Superoxide anion (O2-) production of resident macrophages increased with exposure to spent culture media from macrophages obtained after intestinal reanastomosis (3, 6, 12, 24 hr). This increase reached maximal levels by 6 hr after surgery and thereafter decreased to resident levels by 24 hr after surgery. Exposure of resident macrophages to spent media from cells collected after peritoneal sidewall abrasion (1, 3, 5, 7, 10, 14 days) elevated the production of O2- on Postsurgical Days 3 and 5; however, no effect was observed following exposure to spent media of macrophages harvested on Postsurgical Day 14. Interleukin-1 alpha (IL-1 alpha), transforming growth factor beta (TGF-beta), and tumor necrosis factor alpha (TNF alpha) stimulated phorbol ester-induced O2- production by resident macrophages in a concentration-dependent manner. The secretion of TNF activity by resident macrophages increased following exposure to spent media of macrophages harvested 6 to 24 hr after intestinal surgery. IL-1 alpha, TGF-beta, and TNF alpha elevated the secretion of TNF activity by resident macrophages in a concentration-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412127 TI - Protective effect of the calcium channel blocker diltiazem on hepatic function following warm ischemia. AB - The purpose of this study was to investigate the protective effect of a calcium channel blocker diltiazem (DTZ) on hepatic ischemic injury using a canine model. Hepatic ischemia was induced by temporary clamping of hepatic afferent blood vessels for 60 min with establishment of a portojugular bypass. The administration of DTZ at a dose of 70 micrograms/kg bolus i.v. injection before hepatic ischemia and thereafter via the portal vein at a dose of 10 micrograms/kg/min well restored the hepatic blood flow (HBF, 92 +/- 5%; P < 0.01) and arterial ketone body ratio (AKBR, 1.15 +/- 0.16; P < 0.01) 1 hr after interruption compared with untreated controls (HBF, 61 +/- 7%; AKBR, 0.53 +/- 0.09). In addition, the increase of plasma lactate level and the decrease of mean arterial pressure were significantly suppressed after ischemia (P < 0.05). We concluded that DTZ has a protective effect on ischemia-induced hepatic damage and might be useful in the prevention of primary graft failure caused by warm ischemia in liver transplantation. PMID- 8412128 TI - Effect of hyaluronic acid on rabbit profundus flexor tendon healing in vitro. AB - We performed an in-depth biomechanical evaluation of the effect of hyaluronic acid (HA) on the healing of rabbit profundus tendons cultured in vitro. Seventy eight flexor tendons from 13 rabbits were transected and reapproximated at their Zone II midpoints. Tendons were divided into left and right forepaw groups. Each tendon from the left forepaw group was incubated in one of four possible culture media: control (no HA), low (0.1 mg/ml), medium (0.5 mg/ml), or high (1.0 mg/ml) HA media. HA was added on the first day of incubation. Each tendon from the right forepaw group was cultured in low, medium, or high concentrations of HA, but HA was added after 1 week of incubation in control media. All tendons were cultured for 8 weeks, after which time tenorrhaphies were disrupted and the following biomechanical parameters were determined: apparent maximum stress, apparent strain at apparent maximum stress, normalized energy absorption, and tangent modulus before failure. Comparisons using these parameters showed no statistically significant differences among the various tendon groups. We believe this is the first study of its kind to show no effect of hyaluronic acid on the functional strength of tendon after healing in vitro. PMID- 8412129 TI - Adenine nucleotides of ischemic intestine do not reflect injury. AB - Warm ischemia of the intestine is a medical emergency which results from mesenteric vascular occlusion. In addition, intestinal transplantation techniques will also inevitably result in intestinal ischemia. The recovery of organ function following ischemia depends on the extent of irreversible damage produced by the ischemia and the extent of reflow upon reperfusion. In some organs energy homeostasis has been found to correlate with organ recovery and graft survival following ischemia-reperfusion. Investigating the usefulness of the determination of adenine and pyridine nucleotides as indicators of the extent of ischemic injury in intestinal segments, we found that after an initial 40% decrease in ATP following 30 min of ischemia there was no further decrease despite increasing the ischemia period to 120 min. Similarly, the decrease in NAD+ and NADP which occurred after 30 min of ischemia did not decrease further after 60, 90, or 120 min of ischemia. Xanthine was the only biochemical where an increase appeared to correlate with ischemia duration while energy charge was of no value in indicating injury extent. Additionally, after reperfusion there was at best a poor correlation between recovery of ATP content and the duration of ischemia. Microcirculation reflow after reperfusion indicated ischemia time-related endothelial cell injury. Thus, the measurement of high-energy phosphates in intestinal segments is not of value as an indicator of the extent of intestinal ischemic injury. PMID- 8412130 TI - Prevention of postoperative abdominal adhesions by tissue precoating with polymer solutions. AB - Polyvinylpyrrolidone (PVP) and carboxymethylcellulose (CMC) solutions were evaluated in a rat cecal abrasion model to test the effect of these high molecular-weight hydrophilic polymer solutions on postoperative adhesion formation when used as tissue precoating solutions. Eleven groups of 5-20 animals each were studied including 25 control animals treated with Ringer's lactate (RL) solution. Animals were reoperated at 2 weeks and adhesions were scored according to a 0-4 grading scale. Tissue coating following cecal abrasion failed to inhibit adhesion formation. However, tissue coating with polymer solutions prior to cecal abrasion significantly reduced the formation of post-operative adhesions. Solutions of 1.5% CMC and 5% of a unique gamma-polymerized PVP in RL exhibited the greatest tissue-protective behavior compared to RL controls (P < 0.002). Both CMC and gamma-PVP solutions warrant further investigation as tissue precoatings to inhibit surgical adhesions. PMID- 8412131 TI - Lipopolysaccharide pretreatment of cyclosporine-treated rats enhances cardiac allograft survival. AB - Lipopolysaccharide (LPS), the endotoxin in gram-negative bacterial cell walls, is a major factor in septic shock. Tumor necrosis factor (TNF) appears immediately after LPS release or LPS injection in rats, but when these animals have LPS reinjected for up to 7 days, TNF production is inhibited. Because inhibiting TNF with anti-TNF antibodies prolongs cardiac allograft survival and is synergistic with cyclosporine (CsA), enhanced graft survival could result from inhibiting TNF via LPS pretreatment. Accordingly, heterotopic rat heart transplants were performed in: I, untreated controls: II, LPS pretransplant treatment: III, LPS post-transplant treatment; IV, low-dose CsA post-transplant treatment; V, CsA post-transplant treatment and PBS (LPS vehicle); or VI, LPS pretransplant treatment and low-dose CsA post-transplant treatment, using Brown Norway (BN) donors and Lewis (LEW) recipients. Rejection was defined by a lack of contractions. Results showed that while LPS pre- or post-treatment alone had little allograft survival effect, LPS pretreatment combined with CsA significantly prolonged survival vs control or CsA alone (22.0 +/- 1.6 days vs 6.8 +/- 0.6 days or 13.4 +/- 1.1 days; P < 0.001). Primary MLRs of LPS-pretreated LEW splenocytes cocultured with irradiated BN splenocytes had significantly less [3H]thymidine incorporation than untreated LEW splenocytes (3671 +/- 349 vs 7828 +/- 14 cpm). TNF assays of untreated and PBS-treated LEW spleen cells cocultured with irradiated BN spleen cells had 1.3 and 1.1 pg of TNF/10(6) cells, respectively, in 2 hr, but no TNF from LPS-pretreated LEW cells was detected.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412132 TI - Peritoneal lavage fluid protease levels after in vivo administration of tolmetin in hyaluronic acid. AB - Tolmetin sodium in a hyaluronic acid carrier (tolmetin-HA) was previously shown to reduce adhesion formation and alter the rate and extent to which wound repair cells enter the peritoneal cavity after surgery. In this study, the effect of tolmetin-HA on the levels of protease activities present in lavage fluid from the peritoneal cavity was determined at various postsurgical times. Collagenase activity in peritoneal lavage fluid was suppressed at 6, 12, and 24 hr after administration of tolmetin-HA in comparison to control. Elastase activity levels were biphasic with peak levels at 6 and 72 hr in lavage fluid from controls compared to peak levels at 6 and 48 hr in lavage fluid from treated rabbits. Plasminogen activator activity present in lavage fluid was significantly decreased at 48 hr after surgery in the tolmetin-HA-treated rabbits compared to controls. However, no alteration in the level of plasminogen activator inhibitor activity was observed in either the tolmetin-HA-treated or control rabbits. These data suggest that tolmetin-HA treatment altered the levels of neutral protease activity present in the peritoneal cavity and may therefore effect the proteolytic potential in the peritoneal cavity after surgery. PMID- 8412133 TI - Cushing's reflex and intracranial aneurysm. PMID- 8412134 TI - Diabetic youth: why do they choose suicide? PMID- 8412135 TI - Monotherapy of gram-negative nosocomial pneumonia. PMID- 8412136 TI - Take a left--or--was it a right? PMID- 8412137 TI - The highs and lows of talking to patients about cholesterol. TMA Communications and Public Service Committee. PMID- 8412138 TI - Responding to the health and medical consequences of catastrophic earthquakes. PMID- 8412139 TI - Timing of the breath analyzer: does it make a difference? AB - The purpose of this article is to examine in an emergency room (ER) population the concordance of self-reports of no alcohol consumption prior to injury with breath-analyzer readings in two groups: (1) those patients from whom reports were obtained after they were breath analyzed compared to (2) patients from whom reports were obtained prior to obtaining the breath-analyzer reading. Data were collected on a probability sample of patients attending three health maintenance organization ERs. Among those sampled were 159 patients admitted for initial treatment of an injury, who were breath analyzed within 6 hours of the event and reported no drinking following the event that lead to injury. Of these, 119 were breath analyzed prior to the interview, and none who reported not drinking were positive on the breath analyzer, while of the 37 breath analyzed after the interview, only one was positive who had reported not drinking. Obtaining the breath-analyzer reading following the interview was not found to affect the rate of refusal to provide a breath-analyzer reading; however, it was found to adversely affect obtaining the breath-analyzer reading for other reasons. The data suggest that the concordance of negative self-reports of consumption with breath-analyzer readings remains high in ER populations regardless of when the breath-analyzer reading is obtained; however, it appears best to obtain the reading prior to interviewing the patient for reasons explained below. PMID- 8412140 TI - Evaluation of a saliva alcohol test stick as a therapeutic adjunct in an alcoholism treatment program. AB - This study was designed to evaluate the clinical usefulness of a saliva alcohol dipstick by comparing dipstick saliva alcohol concentrations to simultaneous blood and urine alcohol concentrations. The sample consisted of 211 saliva tests and blood alcohol concentrations and 189 urine alcohol concentrations. The dipsticks had a specificity of 0.965, a sensitivity of 0.895, a positive predictive value of 0.850, a negative predictive value of 0.977 and a Pearson's product moment correlation coefficient of 0.609 for blood alcohol concentrations and salivary alcohol concentrations, suggesting that the saliva dipstick is useful as a qualitative test in certain settings but is less useful than previously reported as a semiquantitative test. PMID- 8412141 TI - Identifying young adult substance abusers: the Rutgers Collegiate Substance Abuse Screening Test. AB - Substance abuse is a serious problem in the young adult population, yet there exists a lack of reliable screening measures for use in identifying problem users in this age group. The Rutgers Collegiate Substance Abuse Screening Test (RCSAST) is a 25-item, true/false questionnaire that was created to provide a reliable means of identifying young adult substance abusers. Three groups completed the RCSAST: a clinical sample of 84 young adult, problem substance users; a group of 33 young adults who were referred to an assistance program but were judged not to have a substance use problem; and a control sample of 87 young adult, nonproblem substance users. The RCSAST correctly classified 94% of the clinical subjects as problem users, and 89% of the control subjects as nonproblem users. The difference between the average total scores for the two groups was highly significant. In addition, the RCSAST was able to distinguish between problem and nonproblem users within the sample of subjects who were referred for evaluation. The findings support the use of the RCSAST in identifying young adult substance abusers. PMID- 8412142 TI - Screening for heavy drinking and alcohol-related problems in young university students: the CAGE, the Mm-MAST and the trauma score questionnaires. AB - The efficacy of the CAGE questions, the Malmo modification of the MAST questionnaire (Mm-MAST) and Skinner's five-item trauma score were used as a part of a larger health questionnaire in a sample of 1,663 female and 707 male first year university students in Finland. The Mm-MAST correlated well (0.51-0.66) to reported alcohol intake and drinking for intoxication, the correlation (0.30 0.39) to CAGE was not as high but significant (p < .001). The trauma score did not correlate significantly to reported alcohol intake. Sensitivity and specificity for Mm-MAST > or = 3 in detecting heavy drinking, defined as a reported consumption of pure ethanol per year of at least 10 kg for women and at least 15 kg for men, was 63.8% and 87.1%, respectively, for female students and 86.3% and 87.4% for male students. Mm-MAST can be recommended as a useful screening tool for heavy drinking and alcohol-related problems in young university students. It is more specific and more sensitive than the CAGE questionnaire in this population. Skinner's trauma score was not useful for the screening of heavy drinking among young university students. PMID- 8412143 TI - Further validation of new scales measuring adolescent alcohol and other drug abuse. AB - The Personal Experience Inventory (PEI) is a recently developed self-report inventory that measures problem severity and psychosocial risk factors associated with adolescent alcohol and other drug involvement. Although previous studies have provided initial support for the psychometric properties of the PEI, this validity evidence was based on uncontrolled studies. The present study examined the concurrent validity of the PEI on a new sample using more controlled research procedures. Results indicated that the PEI Basic Problem Severity scales were significantly related to groups defined by DSM-III-R criteria for substance use disorders and by treatment referral recommendations. Also, the main PEI Problem Severity scale, the Personal Involvement scale, correctly classified a significantly greater proportion of participants into referral subgroups than would be expected given the base rates for the sample. This evidence provides additional support for the validity of the PEI as a problem severity measure of adolescent alcohol and other drug use. PMID- 8412144 TI - Alcohol surveys with high and low coverage rate: a comparative analysis of survey strategies in the alcohol field. AB - Two Swedish alcohol surveys were compared in a search for a reasonable explanation of the large difference in their coverage rates, namely 75% and 28%. In many respects both surveys conducted in the late 1980s by large, well-known institutes, are of a similar type with rather large samples of Swedes. The technique used in the survey with a very high coverage rate (Survey A) takes into consideration the actual drinking pattern of the population studied (i.e., the concentration of drinking on weekends). By dividing a "normal week's consumption" into four units (Monday-Thursday, Friday, Saturday and Sunday), the technique allows one to average periods with varying drinking habits. In the survey with a low coverage rate (Survey B) a "normal week's consumption" was not so divided. A test of internal validity within Survey A underlined the general finding that its higher coverage rate was due to this division. A test of the external validity at aggregate level did not support assumptions about "telescoping" effects in A. Both A and B had a normal week as a basis of measurement for investigating typical drinking habits. The literature concerning differences in coverage rates focuses on the measurement of modal habits versus mean habits. The main explanation of differences is that methods that focus on modal habits (i.e., the Quantity-Frequency Scale) generate a lower coverage rate than do methods that elicit the arithmetic mean (i.e., the last-week recall). Since A and B both belong to the former type of scale, this does not explain our results. PMID- 8412145 TI - Dimensions of adolescent problem drinking. AB - Issues involved in defining and measuring adolescent problem drinking have occupied the attention of alcohol researchers since the mid-1970s. However, appropriate definitions and measures of problem drinking remain important and unanswered conceptual and research questions. The research reported in this study was designed to address these issues by identifying multiple dimensions of adolescent problem drinking and examining correlates of these dimensions using multivariate regression models. A longitudinal sample of 2,771 students who had tried alcohol was used in these analyses. The sample was followed over three waves of data collection, between 1985 and 1990, in middle and high schools located in a southeastern U.S. county. Data from the third wave were used to characterize problem-drinking behavior. Results of exploratory factor analyses supported the conclusion made by others that adolescent problem drinking is a multidimensional phenomenon. Three dimensions measuring (1) level or frequency of alcohol use, (2) problems related to drinking and (3) symptoms of dependency were identified and were only moderately intercorrelated. The independent measures used to model the three problem-drinking measures were the same for each model, and the significant predictors and proportions of variance explained by the predictors differed widely across models. Results of the regression models confirmed the uniqueness of the three measures and suggested that the potency of specific risk factors varies for different types of problem drinking. Prevention implications and recommendations for this area of research are discussed. PMID- 8412146 TI - Sex-role orientation and self-esteem in alcoholic and nonalcoholic women. AB - Theory and research on alcoholism suggest that psychological masculinity and femininity and self-esteem are related to drinking problems in women. In a sample of 60 alcoholic and 60 nonalcoholic women, hypotheses derived from contemporary sex-role theory were contrasted with predictions from a copying styles perspective. Support was found only for the sex-role theory hypotheses. The two groups differed in levels of masculinity, femininity and self-esteem. These differences were reflected in the clustering of the low self-esteem, alcoholic women in the undifferentiated sex-role orientation category. The high self esteem, nonalcoholic women were predominantly androgynous and masculine sex typed. Psychological masculinity was the major factor distinguishing alcoholic from non-alcoholic women, as well as a better predictor of self-esteem than alcoholism. PMID- 8412147 TI - Alcoholism in manic depressive (bipolar) patients. AB - Bipolar patients with alcoholism were compared to nonalcoholic bipolar patients on clinical and family history variables. All patients were systematically examined using a structured interview. There was no increase in the family history of alcoholism in the alcoholic/bipolar. This argues against separate transmission of alcoholism and bipolar illness in bipolar patients who meet diagnostic criteria for both bipolar illness and alcoholism. The possibility remains that alcoholism is secondary to the bipolar illness in some cases. Likewise, there is a possibility that preexisting alcoholism may, by a sequence of events, produce an induced "organic" bipolar picture. PMID- 8412148 TI - EEG, autonomic and subjective correlates of the risk for alcoholism. AB - Electroencephalographic, autonomic and subjective reactions to alcohol were examined among 78 young nonalcoholic men, cross-classified with respect to the presence/absence of a family history of alcoholism (FH) and the presence/absence of a personal history of antisocial personality disorder (ASP). Both an alcohol placebo and alcohol (0.32 ml/kg) were administered in a single laboratory session. The four groups of subjects were compared at baseline, and at several discrete time points before and after consumption of placebo and alcoholic beverages. During the baseline period, ASP+ subjects exhibited significantly more body sway and faster frontal EEG activity than their ASP- counterparts. The combination of ASP with FH was associated, at baseline, with an excessive amount of high frequency (18.6-27.6 Hz) beta activity in the right frontal EEG. After beverage consumption, several significant FH effects emerged that were independent of the effects of ASP. After placebo consumption, FH+ subjects exhibited significantly more fast alpha (10.9-12.5 Hz) activity at the right frontal electrode than FH- subjects. This difference persisted until blood alcohol concentrations began to rise, at which time fast alpha activity in FH+ subjects declined to FH- levels. Differences between the two FH groups were also apparent in their subjective reactions to the placebo and alcoholic beverages. Relative to FH- subjects, FH+ subjects rated themselves as more intoxicated after consuming the placebo but less intoxicated after consuming alcohol. FH+ subjects expressed greater confidence in their ability to resist the offer of an alcoholic drink across most time points. PMID- 8412149 TI - Drinking in various settings as it relates to demographic variables and level of consumption: findings from a national survey in Canada. AB - Based on a large-scale 1989 national survey in Canada, the extent of drinking in different social settings is estimated. Home consumption accounts for the greatest share of total drinking. while drinking in licensed establishments accounts for approximately one-fourth of consumption. This estimate is externally validated against scales data. The set of questions on drinking venues produces estimates of individual's alcohol consumption which have higher coverage rates than the more commonly used quantity-frequency or recent occasion approaches. The distribution of total drinking in different settings is strongly related to demographic variables and individual level of consumption. In particular, drinking in bars and taverns is related to higher levels of drinking and self reported drinking problems. The proportion of drinking in different venues is not strongly related to drinking problems, once demographic variables and individual consumption patterns are taken into account. This does not mean that drinking venue is not a significant factor in the development of problems--it may be via its impact on consumption level and heavy drinking occasions that drinking venue relates to drinking problems. PMID- 8412150 TI - Young adult drinking-driving: behavioral and psychosocial correlates. AB - Behavioral and psychosocial correlates of drinking and driving were examined in two independent samples of licensed drivers aged 18 to 25 selected from the Colorado Division of Motor Vehicles database. Mail questionnaires were returned by 2,300 young adults (1,196 in Sample 1; 1,104 in Sample 2). Structural equation modeling was used to examine the relation of a latent-variable measure of drinking-driving to latent-variable measures of other driving behaviors, problem behaviors and psychosocial variables. Drinking-driving, drug-driving and risky driving were found to comprise a more general, second-order factor of problem driving behavior. Drinking-driving was also found to constitute one aspect of a larger second-order latent variable that included problem drinking, marijuana use, other illicit drug use and delinquent-type behavior. In combination, the variables of problem driving, other problem behaviors, psychosocial unconventionality, risk-taking and hostility/aggression accounted for 57% of the variance in young adult drinking-driving. All of these Sample 1 findings were buttressed by confirmatory analyses in the independent Sample 2 data. The conclusion can be drawn that drinking-driving is part of a more general lifestyle involving behavior and psychosocial unconventionality. PMID- 8412151 TI - A choice-based method to compare alternative alcohol warning labels. AB - Alternative alcohol warning labels were studied with a method used in magical thinking research. In two studies (N = 111, N = 75), subjects completed a questionnaire. A "poison," "toxic" or "causes cancer" label on a beer can was shown to have substantial effects on self-reported choice behavior. In contrast, the alcohol warning label now mandated to appear on alcohol containers did not have nearly as powerful an effect. Explanations for the lack of strong effects of the mandated warning label center around label length, type of warning and salience of the warning. The methodology described in this article appears to test gut-level or conditioned responses to labels, which may be a valuable technique for evaluating warning labels. PMID- 8412152 TI - The social cost of alcohol abuse in Japan. AB - In recent years, alcohol-related health and social problems have become a serious concern in Japan because of the marked increase in per capita alcohol consumption. To facilitate forming a well-balanced policy on alcohol, the economic burden of alcohol abuse on Japanese society was estimated and has been presented in this study. In 1987, the alcohol attributable costs of medical care were estimated at 1,095.7 yen billion, representing 6.9% of the total national medical expenditure. The present amount of lifetime economic value based on age and sex for the Japanese population was calculated using the human capital method to estimate the alcohol attributable mortality costs. In 1987 this cost was estimated at 923 yen billion. By applying a 21% reduction rate, reduced productivity related to alcohol abuse was estimated at 4,257.3 yen billion. Summing up the costs of each component of the overall cost to society, the total cost of alcohol abuse was estimated at 6,637.5 yen billion, representing 1.9% of the gross national product in 1987, which was three times that of the national revenue from taxes on alcohol beverages. Along with the increase in city living, the economic burden of alcohol abuse on our society is expected to rise considerably. Public health intervention urgently needs to be administered in order to reduce the number of alcohol-related health and social problems. PMID- 8412153 TI - Blood activities of antioxidant enzymes in alcoholics before and after withdrawal. AB - Since free radicals and peroxides seem to be involved in the toxicity of alcohol, several authors have examined the variations of blood activities of antioxidant enzymes in alcoholics, but published results are somewhat conflicting. In this study, erythrocyte (E) activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT), and plasma (P) activities of SOD and GPX were measured in 58 male alcoholics without evidence of severe liver disease before and after a 21-day weaning period, and in a control group of 78 healthy men. Before abstinence, E-SOD and E-GPX activities were, respectively, 6.8% and 13.0% lower in alcoholics than in controls (p < or = .05 and p < or = .01, respectively), whereas the slight increases of E-CAT, P-SOD and P-GPX were not statistically significant. After 21 days of abstinence, no change in activities of the erythrocyte enzymes was noticed; conversely, P-SOD activity was reduced by 8.3% (p < or = .01) and P-GPX by 23.3% (p < or = .001). Variations of blood antioxidant enzymes observed in patients were of limited amplitude and do not allow the use of either of them as markers of alcohol abuse. PMID- 8412154 TI - Distribution of GC-subtypes in a series of chronic alcoholics. AB - This article reports on the distribution of GC-(group specific component) subtypes in a series of chronic alcoholics (N = 100). The determination of the phenotypes was carried out by immunoblotting. The results are interpreted and discussed. We observed no association between alcoholism and GC-subtypes. PMID- 8412155 TI - Monoclonal antibodies and their role in modulation of the immune response. AB - Clinical studies in patients having been vaccinated with immunogenic glycoproteins suggest that a monoclonal antibody response is important in helping to induce cell mediated tumor destruction. Such a monoclonal response was noted to characterize the reaction to several different immunogenic antigens administered in an adjuvant setting. The resulting cell mediated reaction appeared to be associated with concomitant antibody dependent cell mediated cytotoxicity (ADCC) and an anti-idiotype monoclonal response. Both 31.1 and 33.28 protein derived colon carcinoma monoclonal antibodies have been found to be capable of turning on natural killer (NK) cell activity as part of ADCC. In vivo studies with specific antigens to induce an anti-tumor response have suggested that the mechanism of tumor cell destruction is complex and associated with T cell activation, anti-idiotype production, and complement binding on the tumor membrane. PMID- 8412156 TI - Characteristics of cancer patients under the age of 20 at medical centers in Israel and the West Bank: a preliminary study. AB - This paper describes the demographic experience on 373 young cancer patients (less than 20 years of age) at two oncology centers initiated in Israel by one medical team in 1975-1977. These units are the Assaf Harofeh Medical Center (AHMC), which predominantly serves a Jewish population (103 cases); and the West Bank Cancer Unit (WBCU), which provides similar care services to the Arab population of the West Bank (270 cases). The two centers have the unique feature of serving two populations residing in close relationship but still differing in many cultural and socioeconomic characteristics. The Arab patients at WBCU tended to be younger than the Jewish children at AHMC. The five most common diagnostic sites for both AHMC and WBCU included hematopoietic system, bones and joints, soft tissue, urinary tract, and brain and nervous system, although not in the same order of occurrence. These tumor sites accounted for approximately 80% of the cases at each center. The experience with WBCU patients was also compared with data for all Jews in Israel. This comparison identified for both sexes combined statistically significant differences in relative frequency of tumors of soft tissue, eye and orbit, brain and nervous system, and thyroid gland. If confirmed by additional data, reasons for the suggested excess risks should be pursued through more definitive epidemiological studies. PMID- 8412157 TI - Stage I testicular tumours: the Tata Memorial Hospital experience. AB - One hundred and fifty-six patients with stage I testicular germ cell tumours--81 seminomas and 75 nonseminomatous tumours--were treated at the Tata Memorial Hospital, Bombay over a 5 year period. Among the seminomas, 71 were treated with post-orchidectomy prophylactic radiation therapy to the retroperitoneum and/or mediastinum, while 10 patients refused radiation therapy and were put on surveillance. The disease-free and total survival in seminomas were 92.6% and 100%, respectively. Among the patients with nonseminomatous tumours, 58 had normal levels of serum biomarkers while 17 had raised biomarkers. In the normal marker group, 20 patients underwent retroperitoneal lymph node dissection (RPLND) with a nodal positivity of 30%, while the other 38 patients refused surgery and were either placed on unplanned surveillance (33 patients) or chemotherapy (5 patients). In this group, the patients undergoing RPLND had a survival rate of 100% as compared to 93.9% in those with surveillance. The overall disease-free and total survival rates in patients with normal markers were 86.2% and 96.6%, respectively. In the raised marker group, 6 patients underwent RPLND with a survival rate of 100% and 11 patients received chemotherapy with a survival of 90%, with the overall survival for patients with raised markers being 94.1%. The overall disease-free and total survival rates for all patients with stage I nonseminomatous tumours were 88% and 96%, respectively. PMID- 8412158 TI - Application of the adenosine triphosphate-cell viability assay in human breast cancer chemosensitivity testing: a report on the first results. AB - Chemosensitivity testing in vitro of breast cancer has been difficult because of small tumour volume, an even smaller yield of viable cells after disaggregation, and the low evaluability rate and sensitivity of current assays. We have employed an alternative approach that quantitates intracellular adenosine triphosphate (ATP) as a measure of cell viability. This ATP-cell viability assay (ATP-CVA) determines in vitro tumor cell viability after exposure to chemotherapeutic agents in comparison to untreated controls following 6 days of incubation. Sixty one fresh breast cancer specimens upon testing yielded an evaluability rate of 95%. Forty-seven of the tumors were untreated primary breast cancers, the remaining 14 were from patients with metastatic disease. Correlations of in vitro drug sensitivity with in vivo response were obtained for 17 treatment regimens in 14 patients with metastatic breast cancer. The level of sensitivity was 90% and the specificity 86%. These preliminary data demonstrated the ATP-CVA to be a practical in vitro approach to breast cancer testing. It will require a larger clinical study for confirmation. PMID- 8412159 TI - Does gastric adenocarcinoma develop after the treatment of gastric lymphoma? AB - Two adult patients with the diagnosis of gastric lymphoma who developed adenocarcinoma of the stomach 8 years after the treatment are presented. Both patients were treated by subtotal gastrectomy followed by irradiation of 4,000 4,500 cGy to the epigastric region and six courses of chemotherapy (vincristine, cyclophosphamide, prednisolone). In our review of the literature, 16 cases of gastric adenocarcinoma following the treatment of gastric lymphoma were found and listed with details. The factors influencing the development of this secondary carcinoma, mainly those treatment related are discussed. The possible role of both radiotherapy and chemotherapy in shortening the latent period for the development of stump carcinoma is emphasized. PMID- 8412160 TI - Sarcoidosis of the pancreas. AB - We report a case of pancreatic sarcoidosis in a patient with radiographic and clinical findings suggestive of pancreatic cancer. Clinically apparent involvement of the pancreas is uncommon in patients with known sarcoidosis and is distinctly unusual as the initial presentation of disease. Sarcoid involvement of the pancreas can mimic adenocarcinoma and should be considered in the differential diagnosis of a pancreatic mass in a patient with symptoms suggestive of sarcoidosis. PMID- 8412161 TI - Pharmacokinetics of cisplatin instilled into the pleural cavity following panpleuropneumonectomy in patients with malignant pleurisy due to lung cancer. AB - The pharmacokinetics of cisplatin instilled into the pleural cavity in patients with malignant pleurisy due to lung cancer were studied. Higher concentrations of total and free platinum in pleural effusion have been maintained for longer than 72 hours in patients who were subjected to panpleuropneumonectomy than in those who received simple drainage. Early rapid drop of total platinum within 6 hours was significant in the drainage group. Total platinum level in the serum of the panpleuropneumonectomy group gradually increased during 1-72 hours, however, that of the drainage group was highest at 1 hour after instillation and declined gradually with time. Free platinum in the serum was also present at lower concentrations in the panpleuropneumonectomy group than in the drainage group. These facts may be ascribed to the absorptive activity of parietal pleura which is present in the drainage group but absent in the panpleuropneumonectomy group. In summary, removal of the parietal pleura by panpleuropneumonectomy could cause cisplatin to be not only more active but also less toxic in patients with malignant pleurisy due to lung cancer. PMID- 8412162 TI - Intraportal injection of monoclonal antibody in nude mice bearing hepatic metastases. AB - Using a model for hepatic human colorectal carcinoma metastases in athymic mice, we compared the selective [intraportal (ip)] and systemic [intravenous (iv)] injection of radiolabeled monoclonal antibody (mAb) strongly reactive against the cell line. Percent injected dose of radiolabeled antibody per gram (%id/g) of tumor or normal tissues was measured at selected time points (up to 5 days postinjection) within 3 dose levels: 0.1, 1.0, and 2.0 micrograms (micrograms). At each dose level, 3-9 animals were studied in each of 3 groups: animals receiving ip injection (group HT-29-15 ip), those receiving intravenous injection (group HT-29-15 iv), and those receiving isotype-matched control antibody via the intraportal route (group BL-3 ip). Significantly greater (P < 0.005) %id/g in tumor was seen in group HT-29-15 ip at all time points and dose levels compared to those in groups HT-29-15 iv or BL-3 ip. However, immediately after injection of mAb, there was no difference in tumor %id/g between groups HT-29-15 ip and HT 29-15 iv at the highest dose level. There was no increase in %id/g of mAb in normal liver and blood after ip injection compared to iv injection beyond day 1. Therefore ip injection resulted in higher tumor to liver and tumor to blood ratios compared to iv (P < 0.005). We conclude that delivery of mAb to hepatic metastases can be enhanced by selective injection; this has important implications in the design of future clinical trials utilizing radiolabeled mAb in the diagnosis and treatment of hepatic metastases. PMID- 8412163 TI - Diagnosis and surgical treatment of small papillary carcinomas of the thyroid gland. AB - The preoperative and pathological findings of 38 cases of small papillary carcinoma of the thyroid gland measuring < or = 10 mm in diameter and 74 cases of large papillary carcinoma measuring more than 10 mm were compared. In the small carcinoma cases, the sensitivity of palpation, ultrasonography, and fine-needle aspiration biopsy (FNAB) was higher than that of computed tomography and soft tissue roentgenography of the neck. Therefore, palpation, US, and FNAB should be instituted as reliable diagnostic methods, but other examinations seem unsuitable as methods for preoperative qualitative diagnosis. No definite correlation was found between the intraglandular dissemination and the size of the carcinoma, and the frequency of metastasis was relatively high even in the small carcinoma cases. If the patient elects to undergo surgery, subtotal thyroidectomy with lymph node dissection should be performed for small thyroid carcinomas as well as large carcinomas. PMID- 8412164 TI - Head and neck synovial sarcomas. AB - Eleven cases of synovial sarcoma of the head and neck are analyzed. Patients presented with a mass either in the parapharynx, pharyngeal wall, or nape of the neck. The tumors ranged in size from 3 to 8 cm. Microscopically, the classic biphasic pattern was seen in 10 tumors. Immunohistochemistry and electron microscopy were useful in the diagnosis of the single monophasic variant of synovial sarcoma. Radical surgery was the mainstay of treatment with post operative radiotherapy for residual disease. Five patients were dead of disease while 6 were alive for periods varying from 9 months to 15 years. The purpose of this presentation is to accrue data on this sarcoma at a rare site, and to highlight the histopathological differential diagnosis, which includes both carcinomas and sarcomas. Treatment decisions would be affected by the histology report. PMID- 8412165 TI - Flow cytometric DNA analysis does not predict the radiochemoresponsiveness of esophageal cancer. AB - The relationship between the DNA pattern and the responsiveness to chemotherapy or chemoradiotherapy has been evaluated in 30 patients with squamous cell carcinoma of the esophagus. In 24 patients polychemotherapy with cisplatin (100 mg/m2 on day 1) and 5-fluorouracil (1,000 mg/m2/24 h, continuous infusion of 120 h) every 3 weeks, was performed. Six other patients received chemoradiotherapy with cisplatin 80 mg/m2 on day 1 and 18.5 Gy (split course). Before treatment, at least three endoscopic biopsies were taken from each tumor and frozen at -85 degrees C. Five patients were excluded from the evaluation, three because of interrupted treatment and two due to unsuitable biopsy material obtained endoscopically. The response rate to the cytoreductive treatment was 40% (10/25). DNA content was analyzed with flow cytometry. Out of 25 evaluable patients, a diploid and aneuploid tumor was present in 8 (32.0%) and 17 (68.0%) patients, respectively. According to the DNA pattern, a major response was observed in 4 of 8 patients with a diploid tumor and in 6 of 17 patients with an aneuploid tumor (P = 0.5). No relationship between the percentage of cells in the S-phase and the response to the cytoreductive treatment was evident. Although a slightly higher percentage of major responses was found in euploid tumors, there is no evidence that flow-cytometric DNA analysis can be helpful in the selection of patients for chemotherapy or chemoradiotherapy. PMID- 8412166 TI - Western blot analysis of glycoproteins bearing Lewis(a) and sialyl-Lewis(a) antigens in human colorectal mucosa. AB - Glycoproteins (GPs) bearing Lewis(a) and sialyl-Lewis(a) antigens (Le(a), sialyl Le(a)) derived from human colorectal carcinomas and their surrounding non neoplastic mucosa (normal mucosa) were analyzed using Western blotting. GPs bearing Le(a) were detected mainly as segmental bands of M(r) 310, 220, 160, and 80 kDa in 80% of the normal mucosa, but these GPs were detected predominantly as broad bands ranging from high to low molecular weight (MW) in 71% of the carcinoma tissues. GPs bearing sialyl-Le(a) were detected only in 23% of the normal mucosa and limited on huge MW bands, i.e., more than 400 kDa, whereas these GPs were detected predominantly as broad bands in 49% of the carcinoma tissues. In the cases with lymph node metastasis, the MW of GPs bearing sialyl Le(a) varied over a wide range and were detected as broad bands, compared with the cases without metastasis. In conclusion, the MW of GPs bearing Le(a) and sialyl-Le(a) in normal colorectal mucosa was different from that in colorectal carcinomas. That is, the MWs of GPs bearing Le(a) varied more in carcinoma tissues, and the GPs bearing sialyl-Le(a) from carcinoma tissues had lower MWs than those from normal mucosa. It was, furthermore, suggested that the increased expression of lower MW GPs bearing sialyl-Le(a) are associated with an increased metastatic potential of the tumor cells. PMID- 8412167 TI - Synergistic antitumor activity of mitomycin C and cisplatin against gastric cancer cells in vitro. AB - The synergistic antitumor activity of mitomycin C (MMC) and cisplatin (DDP) against the gastric cancer cell lines MKN-28 and MKN-45 was assessed in vitro using the MTT assay. The synergism of the two agents was evaluated in terms of the interaction index (I.I.). The sequence of MMC followed by DDP showed higher antitumor activity than the reverse sequence against MKN-28 and MKN-45, and the intracellular concentration of platinum was significantly increased in MKN-45 by preincubation with MMC, suggesting that MMC modulates cellular permeability to DDP or the ability of DDP to intercalate DNA. Since these two antitumor agents show different types of toxicity clinically, i.e., myelotoxicity by MMC and nephrotoxicity by DDP, this combination chemotherapy could be advantageous by providing synergistic antitumor activity without increased toxicity. PMID- 8412168 TI - Usefulness of argyrophilic nucleolar organizer staining for histologic grading of soft-tissue sarcomas. AB - Accurate histologic grading is essential for making a proper therapy decision in soft-tissue sarcomas (STS). The usefulness of the argyrophilic stain for nucleolar organizer region (AgNOR) in assessing the histologic grade of STS has been examined. One hundred and forty-two patients with STS confined to the extremity and trunk were selected. Tumors were classified based on the criteria of Enzinger and Weiss ["Soft-Tissue Tumors." St. Louis: C. V. Mosby, 1983]. In addition, non-specific classification was made based on the shape of proliferating cells occupying more than 50% of the field in the sections such as pleomorphic cell, small round cell, spindle cell, epithelioid cell, myxoid, and unclassified tumors. The mean number of AgNOR dots per nucleus of tumor cells was calculated in 200 cells (AgNOR count). Each category of non-specific classification was divided into a high-count group (< 8 AgNOR count) and a low count group (> 8 NOR). The low-count group showed a significantly better prognosis than the high-count group in small round cell and spindle cell tumors (P < 0.007 and P < 0.0005, respectively). Similar results were obtained in pleomorphic cell tumors, though they were statistically not significant because of the relatively small number of examined cases. Most patients with epithelioid cell and myxoid tumors were in the low-count group. These findings suggest that the assessment of histologic grading of STS could be made effectively by the non specific classification and the aid of AgNOR staining. PMID- 8412169 TI - Surgical management of accessory parotid tumors. AB - Accessory parotid gland tumors are defined as masses within salivary gland tissue located adjacent to Stensen's duct, but separate from the main body of the parotid gland. These tumors usually present as asymptomatic cheek masses. There is a temptation to excise these masses locally; however, the likelihood of injury to branches of the facial nerve is high. The best surgical approach to tumors in the accessory parotid region is via a standard parotid incision and concomitant superficial parotidectomy. Eight patients have been surgically treated with accessory parotid gland masses. Six patients had mixed tumors, one had a low grade mucoepidermoid carcinoma, and one had a localized parotitis. Our approach included a standard parotid incision, raising an anterior flap beyond the mass, and exposing the main trunk of the facial nerve, with careful tracing of all its branches. This approach to accessory parotid gland tumors is superior in that it provides a better margin of resection and minimizes functional and cosmetic deformities. Most importantly, there is less danger of injury to branches of the facial nerve. PMID- 8412170 TI - Primary gastric lymphoma. AB - The records of 30 patients with primary gastric lymphoma and a minimum of 5 years of follow-up were reviewed and clinical and pathologic prognostic factors analyzed. The overall 5-year survival was 40% (median 23 months). No significant relationship between surgical and patient age, sex, duration of symptoms, macroscopic appearance, or size of the primary lesion or degree of serosal infiltration was demonstrated. Stage of disease and site of primary had an impact on prognosis. Survival was improved in patients with stage I-II disease (P < 0.05) and in patients with primary located in the distal third of the stomach (P < 0.05). Although histology in all three classifications did not correlate well with survival, patients with low-grade lymphoma according to Kiel showed improved outcome (P < 0.05). Five-year survival of 11 patients with positive lymph nodes, 6 of whom were treated with cytotoxic therapy, was 54% and comparable to that of 7 patients (56%) with no nodal involvement who did not receive chemotherapy after surgery. PMID- 8412171 TI - Renal cell carcinoma: the size variable. AB - Three hundred and thirty-seven renal cell carcinomas treated at University of Illinois Affiliated Hospitals were reviewed. There was an inverse correlation between tumor size and survival. PMID- 8412172 TI - Expression of cellular oncogenes in human gastric carcinoma: c-myc, c-erb B2, and c-Ha-ras. AB - Expression of oncogenes in gastric cancer tissue was evaluated with immunohistochemical staining methods using monoclonal antibodies to products of the oncogenes. Rates of expression in gastric cancer tissue were 50% for c-myc, 72% for c-erb B2, and 56% for c-Ha-ras oncogenes. Expression of these oncogenes in gastric cancer was not correlated with the histologic differentiation. The c Ha-ras oncogene was positive in 19 of 26 cases with lymph node and/or distant metastasis: the positive rate was significantly higher than in cases without metastasis. Results suggest that c-Ha-ras oncogene is related to the prognosis of gastric cancer. The rate of expression of c-myc and c-Ha-ras oncogenes in gastric cancer tissues was higher in the DNA aneuploid group than in the DNA diploid group. Expression of c-myc and c-Ha-ras oncogenes correlated with other prognostic factors such as DNA ploidy pattern and metastasis. These oncogenes can be used to evaluate prognosis of gastric cancer patients. PMID- 8412173 TI - Intratumor heterogeneity of DNA ploidy and correlations with clinical stage and histologic grade in prostate cancer. AB - Paraffin blocks from 60 patients with prostate cancer were used to study the DNA ploidy patterns by flow cytometry. Nineteen patients had stage A disease, 11 had stage B, 9 had stage C, 20 had stage D, and 1 was of unknown stage. Histologically, 32 of the cancers were well differentiated, 21 were moderately differentiated, and 7 were poorly differentiated. Eighteen patients had an aneuploid or tetraploid (A/T) pattern and 42 had a diploid pattern. Seventy-one percent (5/7) of patients with poorly differentiated, 48% (10/21) with moderately differentiated, and 9% (3/32) with well-differentiated histology had A/T patterns (P < 0.01). Forty-five percent (9/20) of patients with stage D, 44% (4/9) with stage C, 27% (3/11) with stage B, and 5% (1/19) with stage A had A/T patterns (P < 0.05). Nine patients with an A/T pattern also had DNA ploidy studies done on the "benign" part of the specimen. These specimens showed diploid patterns although three of these patients had well-differentiated tumor in the "benign" designated part of the specimen. One patient with mixed histology had an aneuploid pattern on the poorly differentiated section and a diploid pattern on the well-differentiated section of the "malignant" designated part of the same specimen. We conclude that prostate cancer patients with non-diploid tumors have more advanced disease and less differentiated tumors than patients with diploid tumors and that considerable histological and ploidy heterogeneity may be present in different parts of the same paraffin-embedded specimen. PMID- 8412174 TI - Utility of the neodymium:yttrium-aluminum-garnet (Nd:YAG) laser for extensive pulmonary metastasectomy. AB - The neodymium:yttrium-aluminum-garnet (Nd:YAG) laser has been reported to have advantages in pulmonary resections including superior control of bleeding and air leak with relative sparing of pulmonary parenchyma. Extensive metastasectomies were performed in two patients with bilateral metastases who might have been excluded from surgery with other techniques. The patients were discharged within 8 days after uneventful postoperative courses. While this modality shows promise in permitting extensive metastasectomy, only long-term follow-up will demonstrate the ability to provide results comparable to those achieved with previous techniques. PMID- 8412175 TI - Treatment of invasive thymoma with pleural dissemination. AB - The outcome of eight patients with invasive thymoma accompanying pleural dissemination was investigated. Only two patients had mediastinal tumor resection and pleural disseminated tumor excision. Seven patients underwent radiotherapy to the mediastinum and/or disseminated tumors. A clinical response to radiotherapy was achieved in the six patients with evaluable lesions (complete response in five patients and partial response in one). The estimated 5-year survival rate was 87.5%. Four patients were alive more than 10 years. So far, the mediastinal tumors of seven patients have been controlled for periods ranging from 42 to 154 months. Recurrence in six patients appeared as pleural tumors. Four out of the six patients had five courses of radiotherapy to the recurrent pleural tumors, four of which achieved complete response. No distant metastases were observed at any time. These observations suggest that radiotherapy should be the primary mode of treatment in cases of invasive thymoma with pleural dissemination. PMID- 8412176 TI - Biliodigestive anastomosis. PMID- 8412177 TI - Malignant melanoma of the perineum. AB - We conducted a retrospective review of the medical records of 18 patients diagnosed at the University of Colorado with perineal melanoma. Clinical characteristics including Clark's level, recurrence, and survival were studied. Ten patients (55.6%) had vulvar melanoma, 2 patients (11.1%) had vaginal disease, and the remaining 6 patients (33.3%) had primary rectal melanoma. All patients were treated surgically at the time of diagnosis. At a median follow-up of 32.7 months, 15 patients had evidence of recurrence. Nine of these patients are dead and six are alive with disease. The average time to recurrence was 14 months and the most common site was the regional lymph nodes in 10 of the 14 patients (71.4%). This study confirms the poor prognosis in perineal melanoma and the only effective treatment to date is early diagnosis and complete resection. PMID- 8412178 TI - Carotid body tumors. AB - A retrospective study of 20 patients treated at the Tata Memorial Hospital over a period of 50 years, 1941-1991, is presented. Seventy-five percent (15/20) of the patients presented before their fifth decade with a male-to-female ratio of 2:1. The commonest presenting symptom was a painless lump in the neck. Twenty-five percent (5/20) had an incisional biopsy done elsewhere prior to referral to our institute. We performed a subadventitial excision of the tumor in 17 patients, 3 of whom were given postoperative radiation therapy. Three patients underwent complete excision of the carotid system without a vascular replacement, one of whom developed hemiparesis secondary to a cerebral infarct. There was no operative/postoperative mortality. Cranial nerve palsy was seen in 45% (9/20) of patients; the hypoglossal nerve was most commonly affected. One patient had a recurrence, 6 years after surgery, whereas 60% of patients operated on before 1987 have completed a 5-year disease free survival. Forty percent of patients have yet to have a 5-year follow-up period. Reports of newer diagnostic and therapeutic modalities are discussed. PMID- 8412179 TI - Salivary duct carcinoma. AB - Salivary duct carcinoma is an uncommon malignant tumor that occurs mainly in the parotid gland of elderly men. The 11 cases of salivary duct carcinoma which are included in this study occurred in older men (mean age 56 years) and were located in the parotid (7), submandibular salivary gland (2), and the minor salivary glands in the maxilla (2). The maximum tumor dimension ranged from 3 to 9 cm. Microscopically, all had infiltrating margins, with circumscribed groups of epithelial cells arranged in various patterns; the invasive component was embedded in a desmoplastic stroma. Perineural invasion and lymph node metastasis were noted in seven and three cases, respectively, at the time of initial surgery. Radical surgery was offered to ten patients and postoperative radiotherapy to nine patients. Salivary duct carcinoma appears to be an aggressive tumor with distinctive histological features, which has not been described in the minor salivary glands of the maxilla to date. The clinicopathologic features of these tumors are presented, with a review of the literature. PMID- 8412180 TI - Pancreatic ductal cell carcinoma producing pancreatic elastase 1. AB - The elevation of plasma pancreatic elastase 1 in patients with pancreatic cancer is caused by concomitant pancreatitis. There is no report that pancreatic ductal cell carcinoma tissue itself produced elastase 1. A 54-year-old woman underwent total pancreatectomy for pancreatic ductal cell carcinoma. Her plasma elastase 1 level subsequently decreased from the preoperative value of 406 ng/dl to values ranging between 96 ng/dl and 119 ng/dl, until 6 months after the operation, when it began to increase gradually to a maximum of 300 ng/dl in association with local recurrence and hepatic metastasis. Immunohistochemical staining of elastase 1 was positive in the primary tumor resected at operation as well as in the locally recurrent and hepatic metastatic tumors obtained at autopsy. These findings suggested that elastase 1 was produced by the tumor cells in this patient. This is the first case in which pancreatic ductal cell carcinoma cells produced elastase 1. PMID- 8412181 TI - Enteroenteric intussusception due to a metastatic malignant fibrous histiocytoma. AB - Intussusception secondary to metastatic sarcoma is an unusual cause of small bowel obstruction. When a patient who has no history of a previous laparotomy, and has a known malignancy which metastasized hematogenously, presents with small bowel obstruction, the diagnosis of intussusception should be considered. The patient should be evaluated and treated accordingly. PMID- 8412182 TI - Chemoprevention of premalignant lesions of the upper aerodigestive tract. Workshop, February 10-14, 1993, Amelia Island, Florida. Proceedings. PMID- 8412183 TI - Molecular epidemiology of lung cancer and the modulation of markers of chronic carcinogen exposure by chemopreventive agents. AB - Chronic inhalation exposure to environmental carcinogens such as polycyclic aromatic hydrocarbons (PAHs), cigarette smoke, 4-aminobiphenyl (4-ABP), ethylene oxide, and styrene is associated with elevations in biomarkers such as DNA adducts, protein adducts, sister chromatid exchanges (SCEs), chromosomal aberrations, gene mutation, and/or oncogene activation. These biomarkers indicate an increased cancer risk for the exposed population, although quantitative estimates cannot be made with certainty. There is convincing epidemiological evidence that the antioxidant and free radical-scavenging vitamins C and E and beta-carotene (beta-C) protect against cancer of the lung and other epithelial tissues, with somewhat weaker evidence for retinol. Experimental studies demonstrate that these micronutrients are capable of blocking or reducing tumor formation caused by diverse carcinogens. A variety of mechanisms appear to be involved, including suppression of carcinogen activation, enhancement of carcinogen detoxification, induction of cellular differentiation, inhibition of mutagenesis, enhancement of immunologic function, and/or reduction of the formation of carcinogen-DNA adducts, SCEs, micronuclei, and other markers of genotoxic damage. Therefore, we have recently investigated the possible modifying effect of serum vitamins C and E, beta-C, and retinol on a number of such biomarkers in a case-control study of lung cancer, and in a cross-sectional study of heavy smokers. Preliminary results indicate an inhibitory effect of certain vitamins on DNA adduct formation. A significant number of human intervention trials are ongoing involving these vitamins. It appears that biomarkers can provide useful intermediate endpoints for assessment of both the mechanisms and the efficacy of chemopreventive agents. PMID- 8412184 TI - Metabolic activation, DNA adducts, and H-ras mutations in human neoplastic and non-neoplastic laryngeal tissue. AB - Metabolic activation, DNA adducts, and H-ras mutations were examined in human laryngeal tissue (n = 16) from both smoker and non/ex-smoker patients with laryngeal cancer. DNA adducts detected by 32P-postlabelling were evident only in smokers (n = 13); in fact, smoking cessation for as little as 10 months resulted in no DNA adducts detected (n = 3). Total DNA adduct levels in these samples were significantly correlated with levels of cytochromes P-4502C and 1A1 in laryngeal microsomes. Moreover, the P-4501A1 levels represent the highest yet found in human tissues. In contrast, laryngeal microsomes did not have detectable P-4501A2 activity, while laryngeal cytosols showed appreciable N-acetyltransferase activity for p-aminobenzoic acid (NAT1) but not sulfamethazine (NAT2). DNA was extracted from laryngeal specimens and amplified by PCR. Nylon filter dot or slot blots were hybridized with 32P-labelled probes for codons 12, 13, and 61 of the H ras gene. Sixty percent of specimens demonstrated mutations in either codon 12, 13, or 61; a single common and specific mutation was a Gln-->Glu transversion in codon 61. This mutation appeared in 5 laryngeal specimens, all from smokers. These results implicate cigarette smoke components, bioactivated by CYP1A1 and/or CYP2C, in DNA adduct formation. These results also demonstrate a probable smoking related H-ras Gln-->Glu transversion in codon 61. PMID- 8412185 TI - DNA adducts in model systems and humans. AB - The etiology of chemically induced cancer is thought to involve the covalent binding of carcinogens to DNA (adducts) leading to mutations in oncogenes or tumor suppressor genes, and ultimately to tumors. Thus, the DNA-carcinogen adduct has been used as a measurable biochemical endpoint in laboratory studies designed to assess carcinogen exposure, carcinogen metabolism, mutagenesis, and tumorigenesis. Unfortunately, the significance of adducts in the etiology of human cancer is still unclear. This is partially due to the difficulty detecting adducts at carcinogen exposures relevant to humans, which are often orders of magnitude lower than animal model exposures. The relationship between adducts and higher biological effects is also not known at low doses. We have been assessing the DNA damage caused by exposure to heterocyclic amine carcinogens in the diet. Using the technique of 32P-postlabeling in combination with accelerator mass spectrometry, we have determined that DNA adduction in rodents decreases linearly with decreasing dose from the high doses used in typical cancer bioassays to the low doses relevant to human exposures. For a given tissue, adduct levels are correlated with dose, but the level of DNA modification by carcinogens is tissue specific and does not completely correlate with tumor site. This lack of correlation may be due to differences in adduct formation and repair rates among tissues. Comparison of carcinogen metabolism routes between rodents and humans also indicates that species differences could influence the amount and type of damage resulting from exposure to these carcinogens. The use of model systems to study dosimetry, species differences in adduction, and role of adducts in mutation will ultimately lead to a better understanding of the significance of adducts in human disease. This should eventually allow the use of adducts as biomarkers for estimating carcinogen exposure and individual susceptibility. PMID- 8412186 TI - Epstein-Barr virus and nasopharyngeal carcinoma. AB - The Epstein-Barr virus (EBV) has been studied for over 25 years as a probable cause of certain human cancers, including nasopharyngeal carcinoma (NPC). This is a low-incidence head and neck cancer in Western countries (including the USA), but is the third-leading cancer in males in Southeast Asia. Evidence supporting an etiologic relation between this virus and NPC includes the fact that there is a 100% infection rate in patients with this cancer and that EBV DNA and antigens have been demonstrated in all biopsies examined to date. The determination that EBV is at least a major co-factor in the etiology of NPC has led to the development of new diagnostic and prognostic tests for this disease using anti viral markers. Of particular importance to the diagnosis of NPC were the findings, initially reported by the Henles [Int J Cancer 17:1-7, 1976], that the serum of patients with NPC contain IgA antibodies to EBV at a high frequency. In general, 80-90% of patients with this disease contain serum IgA antibodies to EBV as opposed to 10-30% of the normal population. This finding has resulted in the development and successful employment of tests measuring this antibody as adjuncts to pathology in the diagnosis of NPC including the occult form. In addition, this finding has resulted in the development of tests for the early detection of this disease. The IgA test for antibodies to EBV is currently employed in large screening programs in Southeast Asia designed to identify those individuals at risk for the development of NPC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412187 TI - Papillomaviruses in head and neck disease: pathophysiology and possible regulation. AB - Human papillomaviruses (HPVs) are etiologic agents of both benign and malignant epithelial tumors. More than 60 different types of viruses are known, each associated with tissue site and lesion type specificities and differing probabilities of malignant progression. HPVs type 6 and type 11 cause benign papillomas of mucosal squamous epithelium in the aerodigestive tract, with only rare conversion to malignancy. HPV 16 is the most frequently detected HPV in the genital tract, inducing flat lesions with a significant risk of malignant conversion. In the aerodigestive tract, HPV 16 is found only rarely in benign lesions but is detected in 5-20% of squamous carcinomas. In the aerodigestive tract, HPVs frequently cause latent infection, i.e., viral DNA present in tissue but no evidence of clinical or histologic disease. Approximately 10% of the general population may have latent infections. Regulation and activation of latent infections are not well understood, although it is clear that viral functions are tightly regulated by the state of differentiation of the squamous host cell. Control of viral transcription may be the key to prevention of viral activation, and thus control of disease. Among the possible agents under investigation are retinoids, growth factors, anti-sense RNA which interferes with viral expression, and estrogen metabolites. All of these agents modulate either viral expression or cell differentiation or both. It is hoped that in the near future one or more of these agents will be useful in preventing HPV-associated disease. PMID- 8412188 TI - Squamous cell differentiation markers in normal, premalignant, and malignant epithelium: effects of retinoids. AB - Vitamin A and some of its analogs (retinoids) maintain normal differentiation of epithelial tissues by preventing aberrant squamous differentiation of cells in nonkeratinizing epithelia. They can also reverse squamous metaplasia, which develops in vivo during vitamin A deficiency. These effects are the result of the ability of retinoids to suppress the expression of genes associated with squamous differentiation (e.g., transglutaminase type I, loricrin, involucrin, filaggrin, and keratin K1). In addition, retinoids reverse keratinizing premalignant lesions in the oral cavity, and inhibit the growth and squamous differentiation of head and neck squamous cell carcinomas (HNSCCs) in vitro. Nuclear retinoic acid receptors, which function as DNA-binding, trans-acting, transcription-modulating factors, are considered to be the proximate mediators of the effects of retinoids on gene expression and may mediate the re-regulation of aberrant differentiation and growth of premalignant and some malignant cells, thereby suppressing the development of head and neck cancer. PMID- 8412189 TI - Prospective trial evaluating immunocytochemical-based sputum techniques for early lung cancer detection: assays for promotion factors in the bronchial lavage. AB - To confirm the results of a previous report on the use of monoclonal antibodies in immunocytochemical assays of sputums for the early detection of lung cancer, we designed a new prospective trial in an independent clinical trial population. Since well-characterized Stage I resected non-small cell lung cancer patients have a low rate of tumor relapse and a high (1-3%/year) chance of developing a second primary lung cancer, they comprise a very favorable group for conducting an early lung cancer detection trial. The rate of new lung cancer is about 10 fold in excess of a standard "high" risk population of smokers. To optimize the chance for a favorable outcome, all of the technical components for the trial have been systematically evaluated to ensure that optimal procedures are employed. For example, automated immunostaining of the sputum specimens will be performed. Bronchial lavages will be analyzed in a subset of the trial participants to define additional targets for early lung cancer detection. Two markers will be quantitated, including gastrin releasing peptide and peptidyl glycine alpha-amidating monooxygenase activity. These two markers assess the epithelium's capacity to produce growth factors which may be central to the biology of tumor promotion. Since these assays have not been performed in this context before, we attempted to optimize the specimen handling to permit the receipt of the material from a range of collaborating clinical sites in a condition that permits accurate quantitation of these two biomarkers. Efforts to standardize the assay endpoint stimulated the development of computer-assisted methods of immunocytochemical analysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412190 TI - Chemoprevention strategies: the relevance of premalignant and malignant lesions of the upper aerodigestive tract. AB - Patients with premalignant and malignant lesions of the upper aerodigestive tract have historically been the focus of chemoprevention trials within the United States. Experience with this population has formed the basis for trials involving other environmentally induced cancers such as lung and bladder. Given that head and neck cancer patients are at risk for second primary malignancies, prevention strategies can be directed towards decreasing mortality from these metachronous neoplasias. Validity of these strategies, including risk determination, intermediate endpoints, and preventive efficacy of single and combination agents, can be determined. Current limitations in chemoprevention trials involving these patients relate to the sporadic nature of the disease. In fact, the prevalence of oral premalignancy within the United States has not been clearly defined. Individual physician experience with this disease process is limited. Organizational efforts should therefore be directed towards facilitating clinical trials involving dentists, oral surgeons, head and neck surgeons, and other primary health care providers in the community. Risk factors which identify clinically defined normal or premalignant tissue at risk for malignant progression need to be better defined. PMID- 8412191 TI - Oncogene mutations as intermediate markers. AB - Tumors arise through a series of genetic changes which include activation of protoocogenes and inactivation of tumor suppressor genes. It is now possible to identify rare cells containing genetic mutations in an excess background of normal cells. Theoretically, the identification of a clonal population of cells sharing an early genetic marker for malignant transformation would lead to valuable intermediate endpoints and could diagnose premalignant lesions amenable to chemoprevention. Ideally, these genetic changes would be specific point mutations that occur early in the tumor cascade, prior to the development of a clinically significant tumor. To identify these markers, precise histopathologic and genetic tumor models must be described. Early candidate markers include p53 point mutations in squamous cell carcinoma of the aerodigestive tract. PMID- 8412192 TI - TGF-alpha and EGFR in head and neck cancer. AB - The upper aerodigestive tract mucosa of patients who develop squamous cell carcinoma of the head and neck (SCCHN) is predisposed to abnormally regulated growth, evidenced by the high incidence of synchronous and metachronous malignancies. We conducted a series of experiments to show that aberrant regulation of the epithelial growth factor, transforming growth factor alpha (TGF alpha) and its cell surface receptor, the epidermal growth factor receptor (EGFR), contribute to this predisposition. Using molecular biological techniques, we compared the incidence and mechanism of TGF-alpha and EGFR over-production in fresh tumors and histologically normal mucosal specimens from patients with SCCHN and normal, control patients without cancer. In patients with SCCHN, TGF-alpha and EGFR mRNA levels were significantly elevated in both tumor and normal mucosal specimens as compared to levels in control mucosa from non-cancer patients. Neither an enhancement of message stability nor an increase in gene copy number alone accounted for the elevation of EGFR mRNA. Increased production of TGF-alpha and EGFR mRNAs in the histologically "normal" mucosa of patients at risk for a primary or secondary head and neck cancer may serve both as a marker for malignant transformation and as a target for chemoprevention. PMID- 8412193 TI - Premalignant lesions of the upper aerodigestive tract: biomarkers of genetic alterations, proliferation, and differentiation. AB - The normal distribution of cell division in squamous mucosa is in the basal or adjacent suprabasal cell layers. Migration of cells toward the epithelial surface results in cell differentiation, most often expressed by high molecular weight keratin intermediate filaments and components of the cornified envelope, including "involucrin." These latter expressions of terminal differentiation are common in keratinizing dysplasia and invasive squamous cell carcinomas. However, they are less common in the non-keratinizing dysplasias, which fail to express evidence of epithelial maturation. Cell proliferation occurs in or near the basal layer in normal or reactive/reversible hyperplasias. In dysplasia (both keratinizing and non-keratinizing), cell proliferation is observed at all levels of the epithelium. Concomitant with these abnormalities in proliferation and differentiation are nuclear changes characterized by large hyperchromatic nuclei. The enlarged nuclei reflect increased DNA content, as documented by flow cytometry and image analysis techniques. DNA aneuploidy represents a spectrum of genomic alterations reflecting steps toward the progression to invasive carcinoma, which for the most part, have not yet been identified. PMID- 8412194 TI - Tissue autofluorescence as an intermediate endpoint in cancer chemoprevention trials. AB - Intermediate endpoints which reflect responsiveness to chemopreventive agents are needed in clinical trials. A potential problem in the assessment of these endpoints is their requirement for invasive biopsies. Secondly, their expression within the aerodigestive tract is doubtfully uniform. Without clinically definable disease, the potential for biopsy sampling error is significant. Our approach to this problem is the analysis of intrinsic tissue fluorescence. Various cellular components exist within tissues with the innate capacity to emit a characteristic spectral signature when excited at a particular wavelength of light. It can be postulated that cells in varying stages of progression towards malignancy will differ in both the qualitative and quantitative nature of these intrinsic cellular fluorophores which include folate derivatives and cytokeratins, as well as various vitamins and coenzymes. Using current bioptical technology, we have tested the applicability of tissue autofluorescence to chemoprevention studies through various model systems. In an N-nitroso-N methylbenzylamine (NMBA)-induced rat esophageal cancer model, alteration of the 380 nm emission pattern corresponded to disease progression from normal mucosa through dysplasia to invasive cancer. In a multicellular tumor spheroid model, trans-retinoic acid (RA) altered autofluorescent profiles at multiple wavelengths including intensities at 340 nm, 450 nm, and 520 nm. Such RA-induced alterations corresponded to changes in the state of spheroid differentiation. In vivo human studies are in progress which suggest that aerodigestive mucosal neoplasias can be discriminated from normal mucosa by their autofluorescent profile. Because aerodigestive mucosa can be scanned without the need for invasive biopsies, the assessment of tissue autofluorescence as an intermediate endpoint may prove valuable. PMID- 8412195 TI - Development of chemopreventive agents for lung and upper aerodigestive tract cancers. AB - The lung and upper aerodigestive tract (oral cavity, larynx, pharynx, upper esophagus) will harbor the greatest proportion (approximately 20%) of estimated new cancer cases in 1992. The estimated mortality rate is even higher (32%), which is reflected in a 5-year survival rate of only 7% and 12% for esophageal and lung cancer, respectively. Tobacco use appears to remain the major cause of aerodigestive cancers despite efforts at primary prevention--cessation of exposure. Another strategy to decrease this public health problem is secondary prevention or chemoprevention. Cancer chemoprevention is defined as intervention with chemical agents before invasion to halt or slow the carcinogenic process; potential agents may include minor dietary constituents and pharmaceuticals. The main objective of the Division of Cancer Prevention and Control (DCPC), National Cancer Institute, is to develop promising chemopreventive drugs for use in humans. The testing of cancer chemopreventives for efficacy in the clinic differs from that of cancer treatment drugs. Chemopreventive drug trials involve healthy target populations, and the endpoints are reduced cancer incidence or mortality, or increased latency, with no to minimal toxicity. The lung and upper aerodigestive tract represent a unique opportunity for intervention in this setting. Even with cessation of tobacco exposure, the risk of cancer in the entire epithelium remains high for years due to the "field cancerization" effect. Some of the first chemopreventive trials made use of this system due to the availability of a study population with a tissue at demonstrably high risk for malignant progression. Much of the evidence for chemopreventive efficacy is in the oral cavity because of the well-defined epithelial neoplastic progression, the existence of well-established preclinical models, and relative ease of tissue monitoring and sampling. In one of the first randomized trials, Hong and co workers demonstrated that 13-cis-retinoic acid prevents the appearance of second primary tumors in patients previously treated for squamous cell carcinomas of the oral cavity and upper respiratory tract. Even using a high risk population, chemoprevention trials involve large sample sizes, lengthy duration and follow up, and high cost. To circumvent these problems, the use of intermediate biomarkers as surrogate endpoints is being explored. Intermediate biomarkers are defined as biological alterations in tissue (histological, genetic, biochemical, proliferative, differentiation-related) occurring prior to cancer development. In the oral cavity, studies using modulation of a histological intermediate biomarker, dysplastic leukoplakia, as the endpoint have demonstrated response to a retinoid.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412196 TI - Clinical experience with the micronucleus assay. AB - Because of the logistical and practical problems that make cancer prevention trials using cancer incidence as an endpoint virtually impossible to conduct for the majority of cancer types, there is a desperate need for valid intermediate markers of cancer risk to serve as surrogate endpoints in chemoprevention trials. A long and continually growing list of potential markers has been developed in the recent past. Unfortunately very few, if any, of them have been subjected to the usual quality control requirements for a laboratory test before being applied to clinical settings. Modulation of micronuclei frequency has been reported in a number of chemoprevention trials involving the oral cavity, esophagus, lung, and lower GI tract; however, we have focused our efforts primarily on applying the assay to exfoliated buccal mucosal cells, since much of the published data deal with this site, and oral cancer prevention is the theme of one of our chemoprevention trials. After standardizing the definition of a micronucleus by literature review and direct exchange of slides and photographs with other investigators active in the field, we obtained smears from normal subjects, smokers with or without leukoplakia, and tobacco chewers with or without leukoplakia. Our summarized findings follow: (1) Micronuclei represent only one of numerous cytological abnormalities in exfoliated buccal cells that are manifest particularly in tobacco chewers. These include a high frequency of anucleate, binucleate, and multinucleated cells, abnormal shapes and sizes of nuclei, etc.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412197 TI - Chromosomal biomarkers in the clonal evolution of head and neck squamous neoplasia. AB - The biological behavior of any tumor is the result of changes in gene expression caused by mutations accumulated in the carcinogenic process. In squamous carcinoma of the head and neck (SCCHN) there are numerous complex chromosome abnormalities. To make sense of this complexity it is necessary to identify the consistent chromosome changes in a large panel of tumors, to determine if these changes are true representations of the in vivo situation, and to construct a map of gene loci likely to be involved in the development and progression of cancer. Our findings indicate that cultured tumor cells are cytogenetically stable when compared to cells evolving in vivo. We also found that clues to the sequence of events in clonal evolution of individual tumors can be deduced by studying primary and metastatic tumors from the same patient, by examining separate clones within the same tumor, and by analyzing ploidy changes. Furthermore, comparison of the in vitro karyotypes to in situ analysis of the tumor tissue indicates that in vitro cultures are good representations of the in vivo tumor. We conclude that identification of consistent chromosome changes in cultured cells can lead to the loci of genes important in the clinical behavior of individual tumors. The most frequent chromosome abnormalities in SCCHN, found in 40-60% of tumors, are deletions affecting 3p, 5q, 8p, 9p, and 18q. Less common consistent changes found in 30-40% of tumors are gains affecting 3q, 5p, 7p, 8q, and 11q. Preliminary evidence suggests that loss of 18q may be prognostically important and may involve disruption of genes encoding cell adhesion molecules. PMID- 8412198 TI - Blood group antigens and integrins as biomarkers in head and neck cancer: is aberrant tyrosine phosphorylation the cause of altered alpha 6 beta 4 integrin expression? AB - Head and neck cancer is a capricious disease that varies greatly in its clinical behavior. The development of biomarkers that can distinguish between biologically aggressive and indolent tumors has been a long term goal of our laboratories. Predictive markers applicable to biopsy specimens should facilitate clinical management through early identification of patients at greatest risk for early relapse or metastatic spread. Two prominent cell surface markers that we identified by raising monoclonal antibodies to squamous cell carcinomas are blood group antigens and the A9 antigen/alpha 6 beta 4 integrin. Both of these markers are abnormally displayed in squamous cancers of the head and neck and serve as indicators of early relapse. Loss of blood group antigen expression is a stronger single indicator than is overexpression of the alpha 6 beta 4 integrin. However, use of both markers together is a stronger predictive indicator than is either alone. We know little about the function of the blood group antigens in squamous cells except that the mature antigens are associated with differentiation. Similarly, the function of the alpha 6 beta 4 integrin is also not fully understood. Integrin alpha 6 beta 4 is thought to serve as an extracellular matrix receptor, but its ligand has not been confirmed. In resting epithelium, the alpha 6 beta 4 integrin is polarized to the basal aspect of the basal cell as a component of the hemidesmosome, the anchoring structures of the epithelia. This basal polarization is lost in migrating normal squamous cells and squamous carcinomas. Tyrosine phosphorylation of the beta 4 subunit is absent or greatly reduced in malignant cells and this may be a critical signal for subcellular localization of alpha 6 beta 4 and cell anchoring. On the basis of our current experimental results, we postulate that tyrosine phosphorylation of the beta 4 subunit is a reversible signal that regulates cell migration in normal and malignant cells, and may therefore be an important initial event in the metastatic cascade. PMID- 8412199 TI - Early genetic changes during upper aerodigestive tract tumorigenesis. AB - Upper aerodigestive tract tumorigenesis has been hypothesized to represent a field cancerization process with multistep events based on its association with known carcinogens, its frequent associated premalignant lesions, and its multifocal clinical manifestation. To further explore this working hypothesis, we have examined normal tissue and premalignant lesions in the field of tumors for evidence of genetic change. Paraffin sections of head and neck tumors harboring neighboring premalignant lesions were explored for the presence of chromosome polysomies using in situ hybridization and chromosome-specific centromeric probes. Cells exhibiting random polysomy were observed in the premalignant regions near the tumors. The frequency of polysomy in the tumor field increased as the tissue progressed from normal morphology (33%), to hyperplasia (67%), to dysplasia (95%), and to squamous cell carcinoma (96%). These results support the notions of field cancerization and multistep tumorigenesis in the aerodigestive tract. To determine whether the degree of accumulated genetic alterations might serve as a biomarker for risk of developing malignancy, a set of biopsies of oral premalignant lesions (leukoplakia, erythroplakia) were retrospectively chosen for polysomy analysis from two groups of individuals: one group who subsequently developed oral cancer and one group who did not develop oral cancer. Three of the five individuals who showed significant chromosome polysomies in their biopsies subsequently developed oral cancer, whereas only one of eight individuals with little evidence of polysomy subsequently progressed to oral cancer. These results suggest that evidence of generalized genetic change or instability might be useful as a genetic biomarker for risk assessment. PMID- 8412200 TI - Genetic changes in lung cancer. AB - In an attempt to define the type and temporal sequences of somatic genetic changes that precede the onset of invasive lung cancer, and to search for biological markers useful in screening multiple primary tumors of the upper aerodigestive tract, we have performed a cytogenetic and genetic study using normal bronchial epithelium and primary tumor specimens of 68 patients undergoing pulmonary resection for early stage lung cancer, and normal bronchial epithelium of 5 controls with metastatic sarcomas. Of the 68 lung cancer cases, 31 had a single tumor and 37 displayed multiple synchronous or metachronous tumors. Cytogenetic alterations were observed in 59% (23/39) of the evaluable tumor specimens with complex rearranged karyotypes, particularly involving chromosomes 3 (70%), 17 (39%), 11 (26%), 8, 9, 12 (22%), and 7 (17%). Gene alterations were also detected including overexpression of epidermal growth factor receptor (EGFR) in 63% (36/57), HER2/NEU in 21% (12/56), and p53 mutations in 50% (12/24). The overall frequency of genetic changes (any type) in the tumors was 76% (52/68). In the normal bronchial mucosa, we identified a rearranged karyotype in 20% of the evaluable cases (13/63); particularly simple rearrangements involving chromosomes 3p (6 cases), 7 (6 cases), 17 (3 cases), 9, 11 (2 cases), 8 (1 case); as well as overexpression of EGFR in 39% (20/51) and of HER2/NEU in 14% (7/51). The overall frequency of genetic changes (any type) in the normal epithelium was 46% (30/65). The presence of a rearranged karyotype in the bronchial mucosa was associated with a rearranged karyotype in the tumor sample. Other statistically significant correlations were found between histopathologic and clinical features and the occurrence of the different cytogenetic and genetic changes both in tumors and in the normal bronchial mucosa. No genetic abnormalities were found in the bronchial epithelium of the 5 controls. PMID- 8412201 TI - Micronuclei: a potential intermediate marker for chemoprevention of aerodigestive tract cancer. AB - Because they may be used as a quantifiable estimate of the extent of recent DNA injury, micronuclei, extrachromosomal fragments of DNA, are among the most studied potential intermediate markers of cancer chemoprevention. Serial measurements of micronuclei frequency may be easily performed on scrapings from the oral cavity or on bronchial brushings. Assessment of micronuclei frequency and its response to chemopreventive agents has been incorporated into studies of upper aerodigestive tract and lung cancer chemoprevention. These studies have helped define the characteristics of micronuclei and have suggested a role for this test in future chemoprevention studies. Micronuclei frequency has been shown to be increased in the oral and bronchial mucosa of individuals with known carcinogen exposure and is higher at the site of the greatest carcinogen exposure, such as the site where tobacco quids are held, than in grossly normal appearing mucosa. Treatment with chemopreventive agents leads to a reduction in micronuclei frequency. In oral leukoplakia studies, this effect followed treatment with beta-carotene, retinol, alpha-tocopherol, and 13-cis-retinoic acid. The multistep process of epithelial carcinogenesis results from DNA damage and specific genetic events. That micronuclei reflect ongoing DNA injury suggests the hypothesis that long term suppression of cellular genotoxicity, as reflected by a reduction in micronuclei frequency, ultimately leads to a reduction in cancer incidence. PMID- 8412202 TI - Fenretinide (4-HPR) in chemoprevention of oral leukoplakia. AB - A controlled clinical trial has been underway at the Istituto Nazionale Tumori (INT) of Milan since 1988. The goal of the trial is to evaluate the effectiveness of fenretinide (4-HPR) in preventing relapses, new localizations, and carcinomas in patients with benign postoperative diagnoses who have been surgically treated for oral leukoplakias. This paper presents the design and the preliminary results of this study. To date, 137 patients have been randomized, following surgical excision of oral leukoplakia, to receive either 200 mg 4-HPR daily for 52 weeks or no intervention. Twenty local relapses or new localizations have occurred so far in the control group and 9 in the 4-HPR group. Seven patients have interrupted the intervention because of toxicity. No impaired dark adaptation has been observed. We conclude that 4-HPR is well-tolerated and appears to be effective in preventing relapses and new localizations during the treatment period. PMID- 8412203 TI - Beta-carotene and vitamin E in oral cancer prevention. AB - The ultimate proof that a putative chemopreventive agent does prevent cancer is a demonstration of reduced cancer incidence in a targeted population. However, because of practical and logistical considerations, such trials are virtually impossible to conduct for the majority of cancers. Therefore, a conclusion regarding the efficacy of chemopreventive activity is based on consideration of a variety of indirect lines of evidence, including laboratory studies, animal model systems, epidemiologic surveys, intervention trials involving reversal of premalignant changes, and the prevention of malignancies in particularly high risk subjects. Furthermore, the only agents worth testing are those with limited, or preferably, no toxicity, since the final use will be prevention in a generally healthy population. Beta-carotene and vitamin E both fulfill all the criteria for suitable chemopreventive agents; several lines of evidence point toward preventive roles for them in oral cancer. In numerous epidemiologic studies, low intake of beta-carotene has been associated with higher cancer risk. Both intake and supplemental use of vitamin E have been associated with a lowered risk of cancer. Smokers, whose habit is a major risk factor, have lower beta-carotene levels in oral mucosal cells when compared with non-smokers. In several laboratory and animal model systems, including the very relevant hamster cheek pouch model, these agents strongly inhibit oral cavity carcinogenesis. Beta carotene and vitamin E produce regression of oral leukoplakia, a premalignant lesion for oral cancer. This has now been shown in seven clinical trials: five with beta-carotene alone, one with vitamin E, and one with a combination of both.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412204 TI - Carcinogen biomarkers related to smoking and upper aerodigestive tract cancer. AB - Smoking is the major cause of upper aerodigestive tract cancers. Among the many constituents of tobacco smoke, polynuclear aromatic hydrocarbons and tobacco specific nitrosamines are strongly implicated as causative factors for these cancers. The probability that these compounds will induce cancer in a given individual will depend on that person's ability to metabolically activate or detoxify them. Chronic production of DNA damage by these metabolically activated carcinogens is consistent with current concepts of carcinogenesis in which multiple genetic changes, such as activation of oncogenes or inactivation of tumor suppressor genes, appear to be critical. Chemopreventive agents which decrease the level of DNA damage should therefore decrease the risk for cancer. Biomarkers such as carcinogen-DNA adducts, carcinogen-hemoglobin adducts, and urinary metabolites of carcinogens will indicate the amount of metabolically activated carcinogen which may damage DNA in an individual and can therefore be used as an index of risk. Selected biomarkers are discussed in this paper. These biomarkers of internal dose have great potential for application in chemoprevention trials. PMID- 8412205 TI - N-acetyl-l-cysteine. AB - The most commonly used chemopreventive agents in the prevention of oral leukoplakia, head and neck cancer, and lung cancer are beta-carotene, vitamin A, and other retinoids. One of the few chemopreventive agents not in this group and presently being used in a clinical trial is N-acetyl-l-cysteine (NAC). NAC, an antioxidant, is used in EUROSCAN, a European Organization of Research and Treatment of Cancer (EORTC) chemoprevention study in curatively treated patients with oral, laryngeal, or lung cancer. The rationale for choosing NAC is based on a variety of experimental data showing its ability to exert protective effects, including extracellular inhibition of mutagenic agents from exogenous and endogenous sources, inhibition of genotoxicity of reactive oxygen species, modulation of metabolism coordinated with blocking of reactive metabolites, protection of DNA and nuclear enzymes, and prevention of the formation of carcinogen-DNA adducts. NAC has also demonstrated an effect on mutagen-induced chromosomal sensitivity assays, and on anticarcinogenicity in experimental animal models. In addition, preliminary data from EUROSCAN show good compliance in treated patients and a low frequency of side effects. PMID- 8412206 TI - Oltipraz: a laboratory and clinical review. AB - Oltipraz [5-(2-pyrazinyl)-4-methyl-1,2-dithiole-3-thione; RP 35972] is a synthetic, substituted 1,2-dithiole-3-thione previously used in humans as an antischistosomal agent. Cruciferous vegetables (e.g., Brussels sprouts, cabbage) contain several agents, including dithiolethiones, which appear to inhibit carcinogenesis; however, it is unclear which dietary compounds produce the protective effects. Animal studies have demonstrated that oltipraz is a potent inducer of Phase II detoxification enzymes, most notably glutathione-S transferase (GST). Laboratory evaluations have shown that dietary concentrations of oltipraz produce marked inhibition of aflatoxin B1-induced hepatic tumorigenesis in rats. Levels of hepatic aflatoxin-DNA adducts, urinary aflatoxin N7-guanine, and serum aflatoxin-albumin adducts decreased when biliary elimination of aflatoxin-glutathione conjugants increased, thus providing predictive biomarkers that measured a chemopreventive effect. In other animal experiments, oltipraz was found to inhibit chemically induced carcinogenesis in bladder, colon, breast, stomach, and skin cancer models. In addition, oltipraz has been shown to be non-mutagenic, a radioprotector, and a chemoprotective agent against carbon tetrachloride and acetaminophen toxicity. More recent studies in rats suggest that unsubstituted 1,2-dithiole-3-thiones may more effectively inhibit aflatoxin-induced hepatic tumorigenesis and induce electrophile detoxification enzymes. Multiple human clinical trials have been conducted using 1.0-4.5 gram doses of oltipraz over 1-3 days for the treatment of schistosomiasis. Phototoxicity has precluded its use in tropical areas. More recently, a 6 month Phase I trial was completed in which patients with resected colon polyps, or females with first degree relatives with breast cancer, were given oral daily doses of oltipraz at 125 mg or 250 mg. The maximum tolerated dose of oltipraz was < or = 125 mg daily. Grade I/II toxicities included photosensitivity/heat intolerance, GI and neurologic toxicity. Peak plasma concentrations were analyzed by HPLC with wide variability. In another Phase I study, a single oral dose of oltipraz was given to normal volunteers at dose levels of 125, 250, 375, and 500 mg. There was no significant difference in half life (t1/2) between the four dose levels nor in clearance at the 125 and 250 mg levels. Peak oltipraz levels > or = 1.0 microgram/mL were achievable with marked interpatient variability. A series of small trials evaluating single oral doses of oltipraz for up to 28 days (dosing range 1 mg/kg-3 mg/kg/day) also showed a short t1/2 (4.1-5.3 hours), a sustained steady state without variation after a loading dose, and increased serum and urine concentrations with consumption of a high-fat diet.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412207 TI - A review of the use of antioxidant supplements in the treatment of human oral leukoplakia. AB - Over the past twenty years, research into the role of antioxidants in the prevention of cancer has increased dramatically. The use of antioxidant supplements to treat oral leukoplakia has gained acceptance due to the success demonstrated in several clinical trials. This review discusses the role of antioxidants in the development of cancer and their possible use in the treatment of oral leukoplakia. PMID- 8412208 TI - Intraepithelial neoplasia, surrogate endpoint biomarkers, and cancer chemoprevention. AB - Neoplasia is a progression of molecular, cellular, and tissue changes starting with a critical cell mutation and advancing by clonal evolution, involving further multiple mutations and expanding mutated clones. This process is characterized by five general stages: latency, focal growth of normal-appearing but disorganized cells, abnormal-appearing cells (dysplasia), microinvasion, and finally, metastasis. The two driving forces of neoplastic progression in an epithelium are mutagenesis and mitogenesis. These forces frequently occur concurrently, produced by exposure of the epithelium to environmental and endogenous mutagens and mitogens. The major strategy of chemoprevention is to block the effects of both mutagens and mitogens during the early stages of predysplasia and dysplasia. Surrogate endpoint biomarkers (SEBs) are tissue, cellular, and molecular changes that correlate with the later development of cancer. Because of the savings in cost, labor, and time, SEBs are urgently needed to replace the use of cancer incidence reduction as the endpoint for chemopreventive agent clinical trials. The advent of computer-assisted cytometry allows each of the seven basic criteria of dysplasia to be individually assayed as an SEB. Since the dysplastic changes that characterize intraepithelial neoplasia are embodied in the causal pathway to invasive neoplasia, they are already validated as predictors of cancer incidence. More attention should be paid to the quality control of SEB assays, including control of variation in cell composition of tissue samples, assay protocol, instrumentation used, and observer performance. The dose-response relationship between a known chemopreventive agent and the SEB should also be evaluated. The Division of Cancer Prevention and Control, National Cancer Institute, has begun a program to test chemopreventive agents in short-term Phase II clinical trials using dysplasia-based SEBs. The SEBs are assayed, when possible, by computerized cytometry. Trials are being conducted for oral leukoplakia, cutaneous actinic keratosis, superficial bladder cancer, pulmonary metaplasia/dysplasia, cervical dysplasia (CIN III), and adenomatous colonic polyps. PMID- 8412209 TI - Early lung cancer as a potential target for chemoprevention. AB - Carcinoma of the lung is the most common cause of death from cancer in the United States. In considering lung cancer for possible chemoprevention trials, we have analyzed the data collected by the collaborative NCI program on early lung cancer. The data indicate that at least 12 years of study of 80,000 people at risk for lung cancer (adult male cigarette smokers) would be required to establish a 25% reduction in squamous carcinoma of the lung. No intermediate markers of developing lung cancer are presently available to shorten the observation period. It is concluded that a study of the magnitude required is not feasible at the present time. PMID- 8412210 TI - Cytologic diagnosis of oral, esophageal, and peripheral lung cancer. AB - Oral cavity. Most carcinomas in situ of the oral cavity present as red or pink lesions that do not have a keratinized surface. Scrapings of such lesions readily disclose abnormal squamous cells diagnostic of cancer. Scrapings of the keratinized white lesions (so-called leukoplakia) are of no diagnostic value. Dentists, who are most likely to uncover precancerous lesions, are apparently not aware of the diagnostic options based on simple scrape smears. The method is also applicable to follow-up of patients with treated cancer of the oral cavity. Esophagus. Cytologic evaluation of esophageal cancer, initially by washings and subsequently by brushings under endoscopic control, is an established method of diagnosis. The diagnostic results are very good in symptomatic cancer patients and have an accuracy reaching 85-90%. Unfortunately the results of treatment of advanced lesions are very poor, with 5-year survival of only about 5%. Serious efforts at detection of early esophageal cancer started in China in the 1960s, using an abrasive balloon technique which was applied to asymptomatic populations in high risk areas such as Linxian in the Henan province of Central China. The Chinese investigators reported the finding of numerous precancerous lesions of the esophagus classified as carcinoma in situ and as dysplasia. Surgical resection of some of the precursor lesions apparently resulted in a significant drop in the rate of invasive carcinoma, although the statistical results were not convincingly presented. The balloon technique has been tested by us and by others in South Africa and in Transkei, confirming its efficacy in the diagnosis of early esophageal cancer. Peripheral lung. Sputum and bronchial brush cytology may uncover bronchogenic carcinoma in situ and early invasive cancers located in the primary or secondary bronchi. Small, peripheral lung lesions usually do not shed cells in sputum or brushings, and their discovery is usually based on roentgenologic finding. The identity of such lesions can be confirmed in most cases by a transcutaneous aspiration. Most of the peripheral malignant lesions are small adenocarcinomas or epidermoid carcinomas, both resectable by routine surgical procedures. Less commonly, oat cell carcinomas may be observed and these lesions should not be treated by surgery. Benign lesions such as granulomatous inflammation and fungal infections may also be identified by aspiration techniques. The prognosis of the resectable carcinomas varies with their size and the presence or absence of regional lymph node metastases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412211 TI - The hamster cheek pouch carcinogenesis model. AB - The Syrian golden hamster cheek pouch carcinogenesis model is probably the best known animal system that closely compares to events involved in the development of premalignant and malignant human oral cancers. Furthermore, it is one of the most well-characterized models for squamous cell carcinomas (SCCs). However, stages of carcinogenesis (initiation, promotion, and progression) have not been well-defined in this system. Basic understanding of the mechanism(s) of carcinogenesis in this organ is instrumental for the development of new strategies for chemoprevention and early chemointervention. To understand the important early events that occur in the hamster cheek pouch carcinogenesis model, we compared it to the mouse skin model, where a number of critical events have been well characterized. We determined that approximately 60% of the hamster cheek pouch SCCs have a mutation in codon 61 of the Ha-ras gene. We also established a two-stage carcinogenesis protocol in this model using a single dose of dimethylbenz(alpha)anthracene (DMBA) and multiple doses of benzoyl peroxide for 45 weeks. Twenty-five percent of tumors developed with this protocol had the same mutation in codon 61 of the Ha-ras gene, confirming that this mutation, as in the mouse skin model, is initiation-related. We examined the sequential expression of hyperplasia, micronucleated cells, ornithine decarboxylase (ODC) activity, polyamine levels, transglutaminase I activity, epidermal growth factor receptor (EGF-R) levels, keratins, gamma-glutamyltranspeptidase (GGT), transforming growth factor-beta 1 (TGF-beta 1), leukoplakia, and carcinomas induced during carcinogenesis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412212 TI - Esophageal carcinogenesis in the rat: a model for aerodigestive tract cancer. AB - A number of chemical carcinogens have been used to study the process of esophageal carcinogenesis. Among the most prominent of these models is the induction of cancer of the esophagus in the rat by the nitrosamine N nitrosomethylbenzylamine (NMBA). In the rat, tumors can occur within 15 weeks of carcinogen administration. The rat model has been used to investigate the mechanism of action of several chemopreventive agents. Among these, the garlic derived agent diallyl sulfide has been shown to be a specific inhibitor of NMBA metabolism. Other investigators have used the model to seek out the relationship of dietary factors and alcohol in esophageal tumorigenesis. With striking histologic parallels to human esophageal carcinoma, the NMBA model provides useful information to study this cancer. PMID- 8412214 TI - Chain terminator sequencing of double-stranded DNA with built-in error correction. AB - Chain terminator sequencing is usually performed on single-stranded DNA. We propose a technique to determine a DNA sequence by sequencing both strands of a double strand in two four-lane gels. The first set of four lanes will contain a superposition of sequence information from the two complementary strands. The second set of four lanes will contain a different superposition in which one of the two single strands is "shifted" relative to the other. This technique provides redundant information sufficient for error correction--not just the error detection achieved by the standard approach of separately sequencing a single strand and its complement. The information content per four-lane gel is more than 50% higher in this double-stranded approach. Using this technique, the number of chain sequencings--including chain syntheses and gel separations- required to achieve a desired level of accuracy can be reduced by up to a factor of two. Unlike the ordinary single-stranded sequencing, some analysis is required to obtain the single-stranded sequence information from the double-stranded data, particularly in the presence of sequencing errors. This analysis and a computer simulation of double-stranded sequencing in the presence of errors are presented. PMID- 8412213 TI - Lung tumors in strain A mice: application for studies in cancer chemoprevention. AB - Strain A mice develop a high incidence of spontaneous lung tumors during their lifetime. These tumors may be found in some animals as early as 3 to 4 weeks of age, increasing to nearly 100% by 24 months of age. The strain A mouse is also highly susceptible to the induction of lung tumors by several classes of chemical carcinogens and has been used extensively as a mouse lung tumor bioassay for assessing the carcinogenic activity of a variety of chemicals. In addition to its use in carcinogen detection, the strain A mouse lung tumor model has been employed extensively for the identification of inhibitors of chemical carcinogenesis. A number of chemopreventive agents including beta-naphthoflavone, butylated hydroxyanisole, ellagic acid, phenethyl isothiocyanate, phenylpropyl isothiocyanate, phenylbutyl isothiocyanate, phenylhexyl isothiocyanate, indole-3 carbinol, etc., have been shown to inhibit chemically induced lung tumors in strain A mice. In most instances, inhibition of lung tumorigenesis has been correlated with effects of the chemopreventive agent on the metabolic activation and/or detoxification of carcinogens. To date, no chemopreventive agent has been shown to inhibit lung tumorigenesis in strain A mice when administered after the carcinogen, i.e., during the promotion/progression stages of tumor development. Efforts should be made to develop a standardized protocol in strain A mice for evaluating chemopreventive agents as inhibitors of both the initiation and progression stages of lung tumor development. PMID- 8412215 TI - A method for the assessment of the quality of mitochondrial preparations. AB - A new experimental approach was applied to the quantitative assessment of the physiological properties of mitochondria. An analytical function (QUAL) was derived from a linear combination of parameters selected by factor correspondence analysis (FCA). The quality of a mitochondrial preparation can be expressed by two numerical values computed from the function QUAL: a mean value and its variance. These statistical parameters can be used for exploring data from planned experiments. PMID- 8412216 TI - On glucose transport and non-enzymic glycation of proteins in vivo. AB - Non-enzymic glycation is a chance event which may occur whenever a protein is in solution with a reducing sugar. The product of this reaction is a covalently linked glycated protein. Plasma proteins are commonly targets of glycation, particularly in the hyperglycaemic circulation, and yet little is known of the changes to protein function caused by glycation or the catabolic fate of these glycated proteins in vivo. The following article examines the effect of glycation on the catabolism of albumin, the principal glycated protein in plasma, by comparing the catabolisms of radio-labelled samples of rabbit naturally glycated and unglycated albumins in normal and diabetic rabbits. We reason that the relatively small changes (5-10%) to the half-life (T1/2) of albumin and to its distribution to body compartments caused by glycation in vivo may reflect an evolutionary adaptation of albumin (and of all plasma proteins) to resist glycation and, possibly, an accelerated catabolism. Presumably, any increase in the rate of catabolism of a glycated protein followed by its de novo replacement with native (unglycated) protein would create an extra energy demand of the host. In contrast to the vertebrates, the use of non-reducing disaccharides as transport fuels in the circulation systems of insects and higher plants may be an adaptation to allow transport of relatively high concentrations of sugar without the problems caused by glycation. PMID- 8412217 TI - Reptilian and avian ovarian cycles and the evolutionary origin of volant birds. AB - The first birds probably evolved from a line of theropod dinosaurs in the late Jurassic or early Cretaceous. The "trees down" theory proposes that avian ancestors were arboreal, whereas the "ground-up" (cursorial) theory suggests that they were terrestrial, and ran and jumped for prey. We present suggestive evidence from reptilian and avian female reproductive biology that supports the arboreal theory, although neontological evidence can never authenticate a paleontological event. The "law of follicular constancy" is an empirical observation that the number of ovulations per female per ovulatory cycle (instantaneous fecundity, or IF) is the same regardless of the amount of ovarian tissue present. In vertebrates with an IF of two or more, surgical removal of one of the paired ovaries (unilateral ovariectomy, or ULO) leads to a doubling of ovulations from the remaining ovary (compensatory ovarian hypertrophy, or COH), this ovary cycling at the same frequency as it did before surgery. In vertebrates that produce one egg alternately from each ovary (an IF of one), however, ULO leads to a form of COH in which the remaining ovary still ovulates one egg at a time but twice as frequently. In most birds, only the left ovary is present; it ovulates a single egg every 1-2 days until the species-specific clutch size is reached. Inasmuch as this avian ovulatory pattern is similar to an accelerated version of that occurring after ULO in a species that alternates ovulation, we propose that birds evolved from dinosaurs with such a pattern. A great majority of extant reptiles with an IF of one are either anoline or gekkonid lizards, and many of these ovulate several times a year. Furthermore, most species in these tropical groups are arboreal. Even considering phylogenetic constraints within anoline and gekkonid lizards, we propose that correlations of arboreality and an IF of one in these groups are implied as adaptive relationships and represent ecological parallelisms. Therefore, we propose that the ancestors of birds were arboreal. Furthermore, they probably were of small size, as are all lizards with an IF of one. PMID- 8412218 TI - Calculation of movement direction from firing activities of neurons in intrinsic co-ordinate systems defined by their preferred directions. AB - In this work we carried out computer simulations to compare different coding algorithms (the tensor network theoretical approach of Pellionisz & Llinas, 1979; and the weighted average population coding model of Georgopoulos et al., 1986) that were originally devised to recompute vectors of the external world from firing rate responses of neurons of the central nervous system. Georgopoulos and his colleagues (1986, 1988) observed, in electrophysiological experiments, that certain neurons of the primate motor cortex are selective to the direction of arm movement in a three-dimensional space (directional neurons). The discharge rate of a cell is highest with movement in a certain direction (the cell's "preferred direction") and decreases as a linear function of the cosine of the angle between the direction of movement and the cell's preferred direction. They calculated a population vector to predict the direction of arm movement from neuronal responses as a weighted linear combination of the preferred direction vectors using several sets of weighting coefficients. It was implicitly assumed in this approach that if the brain uses such coding, the calculations are carried out by a further layer of neurons. The tensor network theory also gives an algorithm to calculate an external vector in an intrinsic co-ordinate system whose basis vectors are distinguished vectors assigned to the individual neurons based on results of physiological observations. However, it goes beyond providing a simple mathematical formula to recompute an external vector (Pellionisz & Llinas, 1979). It is a promising theoretical solution for the problem faced by sensorimotor systems; how to transform information about the environment, measured by a diverse set of sensors, into appropriate responses executed by multiple muscles acting in concert. As the weighting coefficients used by Georgopoulos et al. in their calculations differed from those used by Pellionisz & Llinas, we show here how they are related. We compared the exactness and robustness of the two approaches in computer simulations assuming that the firing rate responses of individual neurons would change from trial to trial even when the movement direction is the same. We also allowed that different sets of preferred directions were used in different trials mimicking the case when different movement-related directional neurons would be active from trial to trial. In our computer simulations the outcome of the different algorithms were fairly similar. No experimental protocol can be devised in which the activity of all the possible active motor cortex neurons taking part in coding the direction of movement could be simultaneously monitored.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412219 TI - Chemical control of eukaryotic cell movement: a new model. AB - Cellular chemotaxis and chemokinesis play important roles in many biological processes. Most continuum mathematical models for these regulatory mechanisms are based on the model of Keller & Segel (1971 a, b), in which cells respond directly to the local concentration of extracellular chemical. We have developed a new model which reflects the receptor-based mechanisms underlying chemical control of cell motion. Our model consists of three coupled partial differential equations, and we use the Boyden chamber (millipore) assay to compare it with a simpler model based on the Keller-Segel approach. The predictions of our model capture the key qualitative features of the experimental data, whereas the simpler model only does so when appropriate functional forms are chosen for the dependence of the transport coefficients on chemical concentration. Using experimental data on the variation of receptor kinetic parameters with temperature, we use our model to predict the effect of decreasing the temperature on both the "leading front" and "migrated cell" measurements taken from Boyden chamber assays. Our results show that changes in the kinetic parameters play a key role in controlling the temperature dependence of cell chemotaxis and chemokinesis. PMID- 8412220 TI - Noise effects on spike propagation during the refractory period in the FitzHugh Nagumo model. AB - Effects of membrane noise on spike propagation on a nerve fiber are studied on the basis of a stochastic version of the FitzHugh-Nagumo model. The noise varies the propagation speeds of spikes and thus changes the interspike intervals of a spike train during propagation. A kinematic description of spike propagation derives expressions for the changes in the interspike intervals. It is shown that the dispersion relation causes the correlation of the interspike intervals characteristic of a spike train propagating in the refractory period. PMID- 8412221 TI - HIV-1 proteinase as structural model of intercellular transport proteins of plant viruses. AB - Intracellular movement of viral infections of plants requires a virus-encoded protein. Alignment of amino acid sequences of central conserved regions of such proteins produced a sequence profile that resembled that of lentiviral proteinases. The known three-dimensional structure of the proteinase encoded by the human immunodeficiency virus-1 (HIV-1) may serve as a model for the three dimensional structure of the central region of the plant viral proteins. Secondary structures predicted for the plant viral proteins from their amino acid sequences correlate well with those predicted from a proteinase model. In addition, the positions of temperature-sensitive and resistance-breaking mutations in the intercellular transport protein of tobacco mosaic virus are consistent with a structural similarity between the plant viral proteins and the lentiviral proteinases. In a suggested model, the dimeric proteinase-similar domain serves as tether for the attachment of N- and C-terminal domains. The C terminal domain may be an RNA-binding domain. The similarity was used to assign intercellular transport function to a previously unidentified coding region of the genomes of bacilliform DNA viruses. PMID- 8412222 TI - A simple method to forecast good promoters of Escherichia coli by means of doublet frequencies. AB - A training sample (taken offhand) of 14 promoter sequences, 70 or 69 base pairs in length each, with their (relative) promoter strengths given by Deuschle et al. (1986), is used to find a relation between some sequence characteristics and the promoter strength. The analysis is based on the doublet frequencies of purines and pyrimidines in the promoter sequence under study. PMID- 8412223 TI - Metabolic pathway characterization from transient response data obtained in situ: parameter estimation in S-system models. AB - The actual values of internal metabolites and fluxes can be measured by a number of experimental techniques and they provide important information for evaluating the properties of a metabolic pathway in situ. In this paper we propose a strategy to properly exploit this information. The suggested approach permits estimation of a set of parameters on the whole system so that a useful model can be constructed and used to describe its components and systemic properties and to predict its behavior under new conditions. A simulated reference pathway is provided to validate this method and to show its utility in metabolic studies. PMID- 8412224 TI - Friendship and the evolution of co-operation. AB - Recent theoretical research on the repeated Prisoner's Dilemma game (PD) suggests that the evolution of co-operative behaviour depends on the rate at which social groups break apart and new groups form. Long-lasting social groups tend to favour the evolution of co-operation. It is therefore of interest to construct models in which individuals must choose whether to maintain their current social relationships. This is done in the present paper, and the analysis suggests that unco-operative individuals will seek to move quickly from one social relationship to another, while co-operative individuals will seek to maintain relationships with other co-operative individuals. As a result of their differing decisions about changing social relationships, co-operators tend to interact with other co operators to a greater extent than do unco-operative individuals, and this difference is an important determinant of the results of the model. In contrast to the original analyses of the repeated Prisoner's Dilemma, the model presented here allows for the existence of stable equilibria with both co-operative and unco-operative individuals simultaneously present in the same population. It is proved that the position of these equilibria (and hence the frequency of co operators) depends on the size of the various payoffs that define the Prisoner's Dilemma game. In addition, it is proved that co-operation can be maintained at a high frequency so long as co-operators are able to maintain their relationships with other co-operators over long periods of time. This is true even if unco operative individuals are able to move from one relationship to another at a very high rate. PMID- 8412225 TI - Optimality analysis of vascular-tissue system in mammals for oxygen transport. AB - The efficiency of the vascular-tissue system in mammals for oxygen (O2) transport to tissue was evaluated by employing the following simulation models; (i) the spherical tissue model for assessing the maximum tissue mass for which a certain number of capillaries located in the center of each sphere can deliver sufficient O2; (ii) the minimum volume model of the multi-terminal vascular system for estimating the energy cost of blood flow supply to that number of capillaries; and (iii), the efficiency evaluation of the whole system by the ratio (the maximum O2 uptake)/(the energy cost). The computer simulation was carried out by inputting the physiological estimates of tissue O2 consumption rate and cardiac output of mammals in the resting and exercising states, which were calculated from the statistically determined power functions of body weight (the allometric relationship) in the range of 10 g to 500 kg. The results obtained from the data during exercise revealed that the optimum capillary number attaining the maximum efficiency alters nearly in proportion to the body weight and that the total tissue mass corresponding to the optimum capillary number agrees well with the actual tissue mass for virtually all the mammals, while their maximum efficiency remains constant. From these results, it is concluded that any mammal is provided with an equally efficient mass transport system, which is optimized for O2 transport to tissue under the highest metabolic activity, regardless of body weight variation. This may suggest one of the reasons why so many species of mammals with greatly different sizes can exist on the earth. PMID- 8412226 TI - Folding of the MS2 coat protein in Escherichia coli is modulated by translational pauses resulting from mRNA secondary structure and codon usage: a hypothesis. AB - Possible translational pauses within the coat protein of the RNA bacteriophage MS2 were located on the basis of a distribution plot of rare codons and RNA secondary structure. It appeared that the position of certain codon pauses corresponds with the size of some nascent polypeptide intermediates, which have been isolated from MS2-infected cells. Other accumulated polypeptide intermediates seemed to be related to RNA regions, where double-stranded secondary structures occur, which probably impede the movement of ribosomes during chain elongation. We assume that a discontinuous translation rate is designed to allow optimal folding of this (and other) polypeptide(s). PMID- 8412227 TI - Self-organization of genetic coding. AB - The self-organization of genetic coding is studied in a simple model system which uses the products of translation as catalysts for the process. The system studied contains protein molecules chosen from a sequence space of high dimension. Catalysts which assign amino acids to codons are chosen in such a way that a random selection of proteins synthesizes further proteins randomly. Under some circumstances, dictated by the genetic information supplied to the system and the manner in which protein function depends on protein sequence, the state of random synthesis is unstable. The system then evolves spontaneously to a new state in which proteins synthesize further proteins in an ordered fashion, typically executing the rules of a simple code for the assignment of amino acids to codons. For some embeddings of protein functions in the protein sequence space, the domains of stability of the ordered and disordered states are calculated. Computer simulation verify that coding self-organization occurs in a variety of systems of the sort studied. Coding self-organization among catalysts which recognize genetic information is a high order co-operative selection process which provides the link between genotype and phenotype needed for the Darwinian evolution of complex biochemical systems. PMID- 8412228 TI - DNA regeneration in the polymerase chain reaction. AB - This paper is an analysis of a model of the regeneration of damaged DNA during the polymerase chain reaction (PCR) made to estimate the degree to which recombination between similar but not identical sequences ("jumping PCR") compromises the results obtained with the technique. The analysis is applicable to museum or field-weathered specimens and also to the damage, such as single strand nicks, which can occur during storage. Two main results are derived: (i) oligonucleotide priming competes extremely effectively against regeneration derived from priming by overlaps of damaged DNA strands. (ii) Extending result (i), it is shown that jumping PCR is not likely to cause inaccurate sequence determination in standard PCR protocols. One exception arises where it is required to determine the sequence of one specified variant of a piece of DNA which is present in many thousand copies. But, generally, experiments can be performed with confidence that the results are very rarely distorted by jumping PCR. The basic model is extended to investigate how to increase the average length of DNA fragments input to PCR experiments. Increased input of less-damaged material to the PCR can theoretically be achieved by performing several cycles of denaturation, re-association and replication on the sample DNA in the absence of exogenous primers. Experimental work will be necessary to confirm this. However, jumping PCR may occur at substantial frequencies during such pretreatment. This problem cannot be avoided but can be reduced in pretreatment protocols by studying sequences (such as mtDNA or rDNA) where multiple copies are subjected to evolutionary homogenizing processes. PMID- 8412229 TI - A mathematical model for the elimination of carbon monoxide in humans. AB - Carboxyhaemoglobin (COHb) build-up in the blood as a result of exposures to carbon monoxide (CO) affects human beings. It is important to eliminate CO from the blood for treatment and health safety. A mathematical model is proposed to compute COHb level in the blood as a function of post-exposure time as CO is eliminated. The model takes into account molecular diffusion, facilitated diffusion, convection, non-equilibrium kinetics of CO with haemoglobin and the important physiological parameters, such as ventilation rate, blood flow rate and the total volume of blood in the body. Endogenous production of CO in the body is included in the formulation. The resulting coupled system of non-linear partial differential equations with physiologically relevant initial, entrance and boundary conditions is solved numerically. The COHb levels computed from our model agree with those measured experimentally (Pace et al., 1950; Peterson & Stewart, 1970). The half-life of COHb, i.e. the time required for the blood COHb to decrease from the initial level to its half-value is computed. The half-life values of COHb computed from our model are in good agreement with those based on experimental data collected under different physiological conditions (Pace et al., 1950). Also, the results predicted from our model give better approximation to the experimental values than the CFK equation (Coburn et al., 1965). It is found that the rate of elimination of CO increases with the increase of inspired PO2 and ventilation rate. PMID- 8412230 TI - The symmetrical architecture of the genetic code systematization principle. PMID- 8412231 TI - Twenty canonical amino acids of the genetic code: the arithmetical regularities. Part I. PMID- 8412232 TI - Models for growth, decline and regrowth of the dendrites of rat Purkinje cells induced from magnitude and link-length analysis. AB - This study examines Purkinje neurons of rats aged 1, 10, 18 and 28 months to investigate growth and decline in the magnitude of the dendritic tree, i.e. the number of exterior links (terminal segments) per cell. Growth in the mean number of exterior links was observed from 1 to 10 months, decline at 18 months and regrowth at 28 months. At 10, 28, and especially at 18 months, the cell size frequency distribution indicates two groups of cells, one of small and the other of large sized cells. The study also examines the relationship of age to lengths of topologically defined links of various types. For each age group we find that exterior links are longer than interior links (non-terminal or intermediate segments). Analysis of the geometric mean lengths of subtypes of exterior and interior links at maturity (10 months) indicates that they follow a Fibonacci series of link lengths, such that mean lengths of topologically defined types of mean exterior links are either about 13 or 8 microns long, while interior links are about 5 microns long. A sequential growth model for adding exterior links is suggested to illustrate a style of growth which could account for the various mean link lengths and the Fibonacci ratio (1.618) between their lengths. Interior link lengths are also dependent on the generation of exterior links from the sides of pre-existing interior links. If the Strahler branching ratio, Rb, should increase owing to growth of terminals from interior links, then mean interior link length would decline. During a period of regression, mean exterior link lengths become shorter and mean interior link lengths become longer. Changes in mean interior link length are much less affected by changes in Rb during regression than is the situation during growth. Finally, the changes in link lengths dictate that the ratio of mean exterior to mean interior link length increases during growth phases from 1 to 10 and 18 to 28 months, and declines during regression from 10 to 28 months. The lowest values of the ratio of mean exterior to mean interior lengths are found at 1 month. This is the period of most intense growth. During this period, the rate of development of new exterior links outbalances the rate at which the links increase in length. PMID- 8412233 TI - Optimal pulmonary arterial blood pressure. AB - Based on the principle of minimum power, a mathematical model of the functional state of the pulmonary circulatory and respiratory systems is presented. The optimization model minimizes the power expended by the right heart and respiratory muscles. The pulmonary diffusing capacity control mechanism is considered. Mean pulmonary arterial blood pressure and ventilation are determined depending on oxygen transport parameters for man under normal and chronic pathological conditions. Theoretical results are compared with clinical data. PMID- 8412234 TI - An unexpected correlation between cardinal temperatures of microbial growth highlighted by a new model. AB - A new model for the prediction of microbial-specific growth rate as a function of temperature is presented. The four parameters of this model are the three cardinal temperatures (Tmax, Tmin and Topt) and the specific growth rate at the optimum temperature (mu opt). A comparison with three other models was made on the basis of several criteria (simplicity and biological significance of parameters, applicability, quality of fit, minimum structural correlations and ease of determination of parameters). A detailed comparison of a 217-point data set, and an extensive comparison of 47 different data sets show that the new model is better than its competitors. The three cardinal temperatures were found to be independent of mu opt. A very strong and unexpected linear correlation between the cardinal temperatures was observed. The consequences of this biological result are discussed, even though causes remain unknown. PMID- 8412235 TI - Predictions of the dynamics of a polygenic character under directional selection. AB - A quantitative genetic model for the response of the distribution of a single metric trait to directional selection is investigated. Particular attention is paid to the performance of approximations that use only limited information on the initial state of the population, such as the mean, the variance, the skewness and the kurtosis. Selection is imposed according to an exponentially increasing fitness function, populations mate at random, have discrete generations, and all genetic effects are supposed to be additive. Neglecting random drift, qualitatively different predictions for the initial response of a large population that has previously been at a mutation-stabilizing selection balance are derived. These depend on different assumptions about the initial distribution and are compared to the exact dynamics of that model. Linkage disequilibrium can be ignored as long as linkage is not too tight. The mathematical analysis rests on the method of cumulants and of cumulant-generating functions and produces exact equations for the evolution of cumulants in this model. Small populations subject to random drift are shown to respond in a qualitatively different way. The case of small populations is treated, primarily, by Monte Carlo simulations. The consequences of qualitatively different assumptions about maintenance of variation through mutation for the initial response to exponential directional selection are discussed. It is concluded that a significant initial increase in variance is unlikely to be observed in selection experiments if the effective population size is not larger than 500 and if response is caused by additive genes. The present results also apply to weak truncation selection. PMID- 8412236 TI - Invasion and maintenance of alleles that influence mating and parental success. AB - Here we develop a model of the invasion, polymorphism, fixation and extinction analysis of genes that differentially influence the mating and parenting success of the two sexes. The model could have application, for example, to the maintenance of homosexuality in diploid populations. The analysis focuses on systems in which mating success is density-dependent and contrasts the results with the density-independent case. Five situations are analyzed, including cases where male and female parenting success is affected in the same and in different directions by the alleles in question. As part of the analysis, a technique is outlined for identifying boundaries between regions where polymorphisms occur and regions where the alleles under consideration go either to fixation or extinction. The results indicate that, unlike in the density-independent case, in the density-dependent case the degree of dominance of the types of alleles analyzed here critically determines whether such alleles are able to invade a population, exist as a polymorphism, or go to fixation. PMID- 8412237 TI - Theoretical analysis of the hemodynamic variables during sustained mechanical alternans: effect of variations in ventricular filling volume. AB - A comprehensive discrete analysis of the hemodynamic variables during sustained mechanical alternans (SMA) shows that these variables are governed by simple mathematical relations. The analysis shows that the value of the slope created by the two alternating beats on the stroke volume-end diastolic (SV-EDV) plane is gamma = (mu - 1).(1 + beta)/(mu - beta), where mu = SVS/SVW, SVS and SVW denote the strong and weak beats, respectively, in the presence of one contractile state of the beating heart, and the beats associated with the higher and lower contractile states, respectively, in the presence of two alternating contractile states; beta = FVS/FVW, FVS and FVW are the filling volumes following SVS and SVW, respectively. Two separate analyses show that this equation is valid, whether SMA is exhibited in the presence of one or two contractile states, and irrespective of the SV-EDV functional relationship. A criterion based on this slope, in terms of measurable parameters, is described to determine if SMA is caused by variations in EDV and FV. This comprehensive quantitative analysis enables a complete classification of the various types of SMA into subcategories with well-defined features. In particular, this analysis confirms former interesting theoretical consequence that when FV is constant, gamma attains the value of 2, independently of the value of mu or any other factor. PMID- 8412238 TI - Environmental effects on fitness-sets shape and evolutionarily stable strategies. AB - Most models on the evolution of sex allocation and life-history traits are based on the existence of compensations between these traits, and often consider them as linear. With a simple model of physiological response to richness of the environment, we show that not only can these compensations take many different shapes, but also that this shape varies as a function of the resource level. Consequently, evolutionarily stable strategies (ESSs) calculation can give different results for the same two functions according to resource level. Thus, selection can act in different directions depending on the "quality" of the environment. Moreover, genetic variability in the resource allocation strategies is likely to be shown better in intermediate environments, where the proportions of allocation have the most crucial effect on the phenotype. PMID- 8412239 TI - Is morphogenesis an intrinsically robust process? AB - Because morphogenesis is dynamically complex, involving many processes operating over different spatial and temporal scales, it is generally assumed that the stability and repeatability of development depends primarily upon accurate control over parameter values i.e. a precise genetic program. However, it is possible that the dynamic coupling between the different mechanisms involved in development reduces the choices available to the system because of bias in successive symmetry-breaking events. Evidence for such stable cascades in morphogenetic processes is described, and a general argument is presented for morphogenesis as an intrinsically robust process. The evolutionary implications of the hypothesis are considerable. PMID- 8412240 TI - Alternatives to meiosis: the unusual genetics of red algae, microsporidia, and others. AB - At a reductional division, the two alleles at a heterozygous locus segregate into different daughter nuclei. Reductional divisions are therefore vulnerable to genetic elements that cause one cell to attack its sister. This danger can be circumvented by creating uncertainty about when reduction occurs. In conventional meiosis, this is achieved by a sequence of two divisions with crossing over. Either the first or the second division can be reductional for most loci. In some red algae, reduction appears to be spread over more than two divisions. The novel reduction sequence of microsporidia creates maximal uncertainty about the time of reduction in two divisions without crossing over. PMID- 8412241 TI - Computer stimulation of interaction of deacylcortivazol with d(TGTTCT)2. AB - We have obtained models for the interaction of deacylcortivazol (DAC) with the hexanucleotide duplex d(TGTTCT)2 (DNA) by computer simulation technique. The drug was made to interact with the DNA in the major groove and by intercalation mode. It was observed that the classical intercalation model with the drug sitting "end on" between base pairs was not possible. Major groove binding and partial intercalation were the two possible models. In the latter case, the drug enters between base pairs in the "head on" position. The relative merits of these two models vis-a-vis effectiveness of DAC are discussed in the paper. PMID- 8412242 TI - A polaron model for electron transfer in globular proteins. AB - Polaron models have been considered for the electron states in protein globules existing in a solvent. These models account for two fundamental effects, viz, polarization interaction of an electron with the conformational vibrations and the heterogeneity of the medium. Equations have been derived to determine the electron state in a protein globule. The parameters of this state show that it is an extended state with an energy of 2 eV. The electron transfer rate for cyt C self-exchange reaction has been calculated in the polaron model. Reorganization energy, tunneling matrix element and the rate constant have also been estimated. The results are compared with experimental data. The influence of model parameters on the significance of the data obtained has been studied. The potentialities of the model are discussed. PMID- 8412243 TI - Magic and medicinal plants of the Ayoreos of the Chaco Boreal (Paraguay). AB - The Ayoreo is a hunter-gatherer tribe of Amerindians which occupy the central northern part of the Paraguayan Chaco. The whole Ayoreo culture cannot be disassociated from religious beliefs. Disease is considered of supernatural origin and as the result of breaking or disobeying the tabu which regulates existence. A description of the shamanic practices is given to understand better the position of health practices in the Ayoreo culture, particularly the use of medicinal and hallucinogenic plants. Fragments of the Asojna ritual and the methods for becoming shaman; the initiation of the last living shaman, as well as references to the magic powers of the shaman are presented. Diagnosis and treatment included invocations to plant and animal spirits and the use of a few medicinal plants. The plants used as medicine or invoked for healing are presented for the first time. Of particular interest is the identification of two Euphorbiaceae as ritual plants by the Ayoreo. PMID- 8412244 TI - Anticonvulsant effects of extracts of the west African black pepper, Piper guineense. AB - A water extract of the West African black pepper Piper guineense L. was tested for activity against audiogenic seizures in DBA/2 mice, and against seizures induced in T.O. mice by N-methyl-DL-aspartate (NMDLA), pentylenetetrazole (PTZ) and maximal electroschock. Single intraperitoneal doses of the extract produced significant protection of DBA/2 mice against audiogenic seizures. The highest of three doses tested produced 100% and 58% protection at 6 h and 18 h after treatment, respectively. The extract also protected T.O. mice against convulsions induced by NMDLA and maximal electroshock but it had no significant effect on PTZ induced convulsions. The doses of the extract tested did not cause significant impairment of performance of T.O. mice on a rotarod test. The results indicate that the extract of P. guineense has prolonged anticonvulsant activity at doses which do not cause significant CNS depression. PMID- 8412245 TI - Screening of plants used in Argentine folk medicine for antimicrobial activity. AB - Screening of 132 extracts from Argentine folk-medicinal plants for antimicrobial activity has been conducted using a penicillin G resistant strain of Staphylococcus aureus, Escherichia coli and Aspergillus niger as test microorganisms. Cephazolin, ampicillin and miconazole were used as standard antibiotics and concentration-response curves were obtained using the agar-well diffusion method. Boiling water extracts of plant materials were tested and 12 species were active against Staphylococcus aureus, whereas 10 were effective against Escherichia coli and 4 against Aspergillus niger. Tabebuia impetiginosa bark, Achyrocline sp. aerials parts, Larrea divaricata leaves, Rosa borboniana flowers, Punica granatum fruit pericarp, Psidium guineense fruit pericarp, Lithrea ternifolia leaves and Allium sativum bulbs produced some of the more active extracts. PMID- 8412246 TI - Ethiopian traditional herbal drugs. Part II: Antimicrobial activity of 63 medicinal plants. AB - A total of 315 extracts/fractions from 63 traditionally used Ethiopian plants were subjected to antimicrobial screening using known strains of Staphylococcus aureus, Salmonella gallinarum, Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans. The agar plate well diffusion method was used at a sample concentration of 1000 micrograms/ml; it was found that all of the plants showed activity against one or more of the microorganism(s). Direct aqueous extracts from six plants were found to be active against all of the test organisms. These findings confirm traditional therapeutic claims for aqueous dosage forms of these herbs. The relative susceptibility of the test organisms to the five types of extracts/fractions indicated a decreasing rank order of: S. aureus, P. aeruginosa, C. albicans, S. gallinarum, E. coli, K. pneumoniae and P. vulgaris. PMID- 8412247 TI - Pharmacological and chemical screening of Byrsonima crassifolia, a medicinal tree from Mexico. Part I. AB - Leaf and bark extracts of Byrsonima crassifolia displayed concentration dependent, spasmogenic effects on rat fundus in vitro and biphasic effects on rat jejunum and ileum in vitro. Dose-related in vivo effects in intact rats using hippocratic screening were: decrease in motor activity, mild analgesia, back tonus, enophthalmos, reversible palpebral ptosis, ear blanching, Robichaud positive, catalepsy (awake) and strong hypothermia. Rat fundus in vitro was used as the bioassay to carry out an activity-directed separation. Bioactive material was concentrated in a 2% acetic acid leaf extract (HOAcE). Potency of HOAcE was increased by the presence of pargyline in the bathing solution. HOAcE was antagonized noncompetively by 1(1-naphthyl) piperazine (1-NP) and cyproheptadine and antagonized competitively by atropine (ATR). Cumulative concentration response curves of HOAcE and serotonin (5-HT) did not show significant departure from parallelism (P > 0.1) and 5-HT potency was 6040 times that of HOAcE (95% confidence limits: 4620-7850). Solvent extraction of HOAcE split the spasmogenic activity of HOAcE into two types: (i) high-efficacy, low-potency, n-butanol extracted, pargyline- and 1-NP-sensitive, ATR-insensitive activity, and (ii) low efficacy, high-potency, ethyl acetate-extracted, pargyline-insensitive, ATR- and 1-NP-sensitive activity. HOAcE lacked muscarinic and nicotinic effects on rat jejunum and frog rectus abdominis. Results suggest the presence of more than one spasmogenic compound in the plant. PMID- 8412248 TI - Plants used in traditional medicine in eastern Tanzania. VI. Angiosperms (Sapotaceae to Zingiberaceae). AB - Fifty-three Angiosperm species (Sapotaceae to Zingiberaceae), which are used by traditional healers in five regions of Eastern Tanzania (Coast, Dar es Salaam, Kilimanjaro, Morogoro and Tanga) are listed. For each species, the botanical name, vernacular name, collection number, locality, habit, geographic distribution and medicinal uses are given. Additionally, information from the literature on medicinal uses, chemical constituents and pharmacological effects are also provided. Data analysis and conclusions to cover all the six parts of the papers previously published are presented. PMID- 8412249 TI - Right gastroepiploic artery used for coronary artery bypass grafting. Evaluation of flow characteristics and size. AB - Questions remain concerning the physiologic capabilities of the right gastroepiploic artery as a bypass graft in the clinical setting. Our last 90 consecutive pedicle right gastroepiploic artery grafts were prepared with intraluminal papaverine and verapamil. Our series comprised 81 male and 9 female patients with average body surface areas of 1.92 m2. Ages ranged from 11 to 79 years (mean 57.2 years). A second to fourth revascularization was undertaken in 32 patients (35.5%). The following arteries were bypassed: posterior descending artery, 63; right coronary artery, 23; distal right, 4; circumflex, 2; left anterior descending, 1; and diagonal, 1. Free flow rates ranged from 42 to 660 ml/min (mean 179.96 ml/min). Internal diameters measured 1.5 to 4.0 mm (mean 2.20 mm) at the anastomotic sites. Pedicle lengths ranged from 16 to 26 cm (mean 19.2 cm). Inotropic support was required in 11 patients (12%) and had no adverse effects on right gastroepiploic artery grafts. There were 2 hospital deaths (2.2%). Angina has recurred in 6 patients. One patient with cardiomyopathy required transplantation 2 years after coronary bypass grafting. Repeat angiography showed widely patent grafts in 18 patients and generalized narrowing in 4 grafts. In only 2 patients of our total experience has right gastroepiploic artery grafting been aborted because of inadequate conduit size. One right gastroepiploic artery had visible atherosclerosis. This study shows that distal right gastroepiploic artery sizes are comparable with sizes of target coronary arteries. However, neither flow nor size is as consistent when compared with internal thoracic artery grafts. Higher flow rates are related to graft anatomic characteristics and larger body surface areas. Spasm, secondary to harvest in these vasoreactive grafts, can be managed appropriately by intraluminal vasodilating drugs. However, use of the right gastroepiploic artery should be avoided in a setting with possible competition of flow. PMID- 8412250 TI - Presence of calcium in the vessel walls after end-to-end arterial anastomoses with polydioxanone and polypropylene sutures in growing dogs. AB - The presence of calcium in the vessel walls after end-to-end arterial anastomoses performed with polydioxanone and polypropylene interrupted sutures was studied in 140 anastomoses in 35 10-week-old German shepherd dogs. Histologic examination with hematoxylin and eosin, van Gieson, and von Kossa staining techniques was performed after the animals were killed 6 months after the operation. Ketamine hydrochloride was used as an anesthetic agent. At the start of the investigation the dogs weighed 14.5 +/- 2.6 kg (mean +/- standard deviation, n = 35), and after 6 months they weighed 45.3 +/- 3.1 kg (mean +/- standard deviation, n = 35). The diameter of the sutured arteries in the first operation was 2.6 +/- 0.5 mm (mean +/- standard deviation, n = 140). With each dog, both brachial and both femoral arteries were used--one artery for each different type of suture. In different dogs, different arteries were used for the same type of suture. The prevalence of calcifications after 6 months was determined from the numeric density of calcifications with standard stereologic techniques. The sutured and sutureless parts taken from longitudinal sections from each artery were studied, and t test values were calculated as follows: In paired samples, statistically significant differences in numerical density of calcifications were seen between sutured and sutureless arterial parts for both materials (sutureless part versus part with polydioxanone sutures, p < 0.001, n = 70; sutureless part versus part sutured with polypropylene sutures, p < 0.01, n = 70); however, in independent samples no statistically significant differences in numerical density of calcifications were seen between the polydioxanone and polypropylene groups for sutured (p > 0.05, n = 70) and sutureless parts (p > 0.05, n = 70). PMID- 8412251 TI - Late functional results after surgical closure of acquired ventricular septal defect. AB - To assess the longer term outlook for patients who have undergone surgery for acquired (postinfarction) ventricular septal defect, we interviewed and studied 60 survivors from a single regional cardiac center between 3 and 144 months after the operation. Including the patients who died within 1 month of the operation, the 5-, 10-, and 14-year survivals (with standard errors) were 69% (65% to 74%), 50% (44% to 57%), and 37% (27% to 46%). Eighty-two percent of patients were in New York Heart Association class I or II. Ten patients (17%) had a persisting but not hemodynamically significant ventricular septal defect. Mean left ventricular ejection fraction was reduced at 0.39 (standard deviation 0.15), but this did not correlate with either New York Heart Association class or exercise tolerance. Twenty-eight patients (47%) had asymptomatic arrhythmias (17 with ventricular premature beats). Angina and other medical problems were not prevalent. PMID- 8412252 TI - The Omnicarbon tilting-disc heart valve prosthesis. A clinical and Doppler echocardiographic follow-up. AB - From 1986 to 1990, 172 patients with a median age of 60.5 years (range 20 to 79 years) received 187 Omnicarbon valves (109 aortic valve replacements, 48 mitral valve replacements, and 15 double valve replacements). Patients were followed-up for a median observation period of 2.5 years (range 4 months to 5.2 years) by clinical and Doppler echocardiographic examination. Follow-up was complete in 98%. Operative mortality (death within 30 days) was 1.7%, and linearized late mortality was 2.6% per patient-year, corresponding to an actuarial survival rate for operative survivors of 89% after 4 years. The overall 4-year postoperative survival was 87% (93% for aortic valve replacement, 77% for mitral valve replacement). Compared with age- and sex-adjusted Swiss death rates, there was an excess mortality of 5% after 4 years. Percentages for freedom from valve-related complications at 4 years are as follows: thromboembolism, 98% (aortic valve replacement, 98%, and mitral valve replacement, 96%); anticoagulant-related hemorrhage, 95%; valve endocarditis, 96%; reoperation, 96%; and permanent valve related impairment, 99%. The overall 4-year event-free survival was 76% (80% for aortic valve replacement and 69% for mitral valve replacement). New York Heart Association class improved in 88% of the patients by 1 to 3 grades, and only 3% remained in class III after operation. For the most commonly used aortic valve (23 mm), Doppler echocardiography revealed a peak pressure gradient of 29 +/- 10 mm Hg, a fractional shortening/peak pressure gradient ratio of 1.34 +/- 0.61, and a performance index of 0.35 +/- 0.08. In the most commonly used mitral valve (27 mm), the mean pressure gradient was 4.0 +/- 2.1 mm Hg. We conclude that excellent clinical and hemodynamic results can be obtained with the Omnicarbon prosthesis, in both the aortic and mitral positions. PMID- 8412253 TI - Vanishing De Vega annuloplasty for functional tricuspid regurgitation. AB - Annuloplasty is performed for significant functional tricuspid regurgitation even if it is presumed that in some cases the regurgitation will regress spontaneously after correction of the left-sided lesion. In an attempt to avoid the drawbacks of a permanent annuloplasty, we used a reabsorbable De Vega annuloplasty in a selected group of patients. Of 73 patients with functional tricuspid regurgitation operated on between May 1989 and May 1991, 25 with pulmonary arteriolar resistance below 400 dyne.sec.cm-5 underwent a De Vega annuloplasty with 2-0 polydioxanone suture. The diagnosis of significant functional tricuspid regurgitation (mean 2.74 +/- 1.05) was established by transthoracic color Doppler echocardiography in all patients. The degree of functional tricuspid regurgitation and pulmonary arteriolar resistance were measured with the patients anesthetized. In 16 patients the regurgitation remained severe (3+ to 4+) and in 9 it was moderate (2+). Twenty-three patients had mitral (12 repairs, 11 replacements) and 9 had aortic (4 repairs, 5 replacements) valve operations. The immediate postbypass residual functional tricuspid regurgitation was 0 to 1+ in 23 and 0 in 2. There was 1 (4%) operative death. The maximum follow-up period was 24 months (mean 13.9 months). There were 2 (8.3%) late deaths. Six patients underwent reoperation because of mitral dysfunction. Four of them who were reoperated on between 2 and 5 weeks after the initial procedure showed no recurrence of functional tricuspid regurgitation. The other 2, reoperated on at 5 and 10 months after the first operation, had recurrence of functional tricuspid regurgitation. Visual inspection of these two tricuspid valves showed a dilated anulus with otherwise normal valves. All surviving patients are in New York Heart Association functional class I or II without significant functional tricuspid regurgitation (mean 0.78 +/- 0.56). We concluded that functional tricuspid regurgitation in patients with low pulmonary arteriolar resistance can be adequately treated by a vanishing De Vega annuloplasty, which will stent the tricuspid anulus for about 4 months. PMID- 8412254 TI - The donor lung: infectious and pathologic factors affecting outcome in lung transplantation. AB - The prevalence of posttransplantation pulmonary infection and the importance of this complication with respect to morbidity in patients undergoing lung transplantation is significant. Over a 1-year period, case histories of all patients undergoing lung transplantation at Barnes Hospital, Washington University, were reviewed to examine the importance of organisms isolated in the donor lung in the development of subsequent invasive infection in transplant recipients. Twenty-eight of 29 bronchial washings (97%) taken from donors before retrieval grew at least one organism. The most common organisms identified were Staphylococcus and Enterobacter. In 12 of these cases (43%), similar organisms were isolated from the tracheobronchial tree of the recipients, and 6 of these recipients (21%) subsequently had invasive pulmonary infections as a result of the organism originally isolated in the donor. We recommend that antibiotic coverage in transplant recipients should be initiated on the basis of Gram stain results and modified on the basis of cultures obtained from the donor lungs. Pathologic analysis of donor lung tissue taken before transplantation was available in 12 cases. Four donors had histologic evidence of established pneumonia in the donor lung, and infections then developed in the recipients. One other patient who received a lung that had widespread bone marrow emboli and subsequent infarction later had a complete anastomotic dehiscence. An additional patient had profound early donor lung dysfunction without any evidence of rejection or infection. Pathologic findings from the donor in this case demonstrated preexistent acute vasculitis with emboli. We suggest that as preservation techniques improve, the opportunities for closer scrutiny of donor lung tissue before implantation will become increasing desirable and feasible. PMID- 8412255 TI - Clinical experience with cardiac retransplantation. AB - Although more than 560 patients worldwide have undergone cardiac retransplantation, few studies of this population have been reported. To evaluate the risk of cardiac retransplantation and to better establish selection criteria, we reviewed the records of all patients who underwent retransplantation at the Columbia-Presbyterian Medical Center. Of 431 patients who underwent transplantation between February 1977 and March 1991, 408 underwent the procedure in the era of cyclosporine-based immunosuppression. Thirteen of these 408 patients underwent retransplantation (including one patient who received a third graft). Indications for the 14 retransplantations included transplant coronary artery disease (n = 8), rejection (n = 5), and intraoperative graft failure (n = 1). Immunosuppression and follow-up protocols used in this cohort were similar to those in the primary transplantation population. No significant differences were found in either actuarial survival between primary transplant recipients (75.1% +/- 2.2% at 1 year and 71.3% +/- 2.4% at 2 years) and patients who underwent retransplantation (71.4% +/- 12.1% at 1 year and 59.5% +/- 14.8% at 2 years) or in linearized rates of rejection and actuarial freedom from rejection between the two groups. No differences between these groups were found with regard to age, sex, race, origin of end-stage heart disease, or early (< 30 day) mortality. The origin of primary graft failure did not correlate with survival outcome in the retransplantation cohort. Follow-up time for patients having primary transplantation ranged from 0 to 8 years (mean 24 months) with a cumulative patient follow-up of 830 patient-years; follow-up time for patients who underwent retransplantation ranged from 0 to 3 years (mean 8.1 months) with a cumulative patient follow-up of 9.5 patient-years. Approximately 50% of patients in both groups had at least one rejection episode by 3 months. Within the limited time period studied after retransplantation, only one patient had transplant coronary artery disease, approximately 27 months after her first retransplantation procedure for acute rejection. These results indicate that the prognosis for patients undergoing cardiac retransplantation is good for patients for whom the indication for retransplantation is identified more than 30 days after initial transplantation. PMID- 8412256 TI - Experiments in cardiac xenotransplantation. Response to intrathymic xenogeneic cells and intravenous cobra venom factor. AB - Permanent tolerance to an experimental cardiac allograft can be achieved by pretransplantation intrathymic inoculation of donor-specific lymphoid cells. We studied the effects of intrathymic inoculation of xenogeneic cells and intravenous cobra venom factor in a rodent model of cardiac xenotransplantation. Lewis rats underwent intraabdominal heterotopic heart transplantation with Syrian hamster donors. In untreated animals, mean graft survival time was 3 days. Five rats had 1 ml of antilymphocyte serum administered intraperitoneally. One day later, 2.5 x 10(7) hamster spleen cells were inoculated into the thymus under direct vision. Twenty-one days after antilymphocyte serum was given, heterotopic heart transplantation with a hamster donor was carried out. In all cases, rejection was accelerated and occurred between 20 minutes and 1 day after transplantation. Mean graft survival time was 5.2 hours (p < 0.0001 versus control). Six animals treated with antilymphocyte serum and intrathymic xenogeneic cells had 0.5 ml of cobra venom factor, a complement antagonist, administered intravenously 3 hours before transplantation and every other day thereafter. Mean graft survival was 3 days, which was not different from the response of naive animals. Animals treated with antilymphocyte serum only had no prolongation of graft survival (mean survival time 3 days, p = not significant). Animals treated with cobra venom factor alone (n = 5) before transplantation and on alternate days subsequently had mild graft prolongation with a mean survival time of 4 days (p = 0.0133). In contrast to experimental allograft models, intrathymic inoculation of xenogeneic cells produces hyperacute rejection in these naturally concordant species. The administration of cobra venom factor abrogates the hyperacute response, but the combination of cobra venom factor and intrathymic inoculation does not produce long-term graft survival. PMID- 8412257 TI - Validity of cardiac output measurement by the thermodilution method in the presence of acute tricuspid regurgitation. AB - Evaluation of patients with acute tricuspid insufficiency may include assessment of cardiac output by the thermodilution method. The accuracy of estimates of thermodilution-derived cardiac output in the presence of tricuspid insufficiency has been questioned. This study was designed to determine the validity of the thermodilution technique in a canine model of acute reversible tricuspid insufficiency. Cardiac output as measured by thermodilution and electromagnetic flowmeter was compared at two grades of regurgitation. The relationship between these two methods (thermodilution/electromagnetic) changed significantly from a regression slope of 1.01 +/- 0.18 (mean +/- standard deviation) during control conditions to a slope of 0.86 +/- 0.23 (p < 0.02) during severe regurgitation. No significant change was observed between control and mild regurgitation or between the initial control value and a control measurement repeated after tricuspid insufficiency was reversed at the termination of the study. This study shows that in a canine model of severe acute tricuspid regurgitation the thermodilution method underestimates cardiac output by an amount that is proportional to the level of cardiac output and to the grade of regurgitation. PMID- 8412258 TI - Contracture of the newborn myocardium after prolonged prearrest cooling. AB - Profound hypothermic circulatory arrest is frequently used to facilitate the surgical repair of congenital heart defects in neonates. Deep hypothermia is achieved by a period of core systemic cooling during cardiopulmonary bypass before cardioplegic arrest. There have been conflicting reports with respect to the consequence of perfusing a nonarrested newborn heart under hypothermic conditions. This in vitro study was designed to prolong the clinically simulated hypothermic perfusion sequence into an extreme condition and to test the hypothesis that prolonged cold perfusion of the nonarrested newborn myocardium could, in fact, be detrimental. Twenty-four newborn piglets (5 to 7 days old) were randomly assigned to four groups and studied in a crystalloid perfused Langendorff heart model. The first two groups of hearts (n = 6 per group) were subjected to either 30 minutes (group I) or 90 minutes (group II) of cold perfusion at 15 degrees C, followed by 90 minutes of ischemia and then 30 minutes of normothermic reperfusion. In a second experiment, group III hearts subjected to 30 minutes of cold perfusion were compared with group IV (90 minutes of cold perfusion) without ischemic insult in either case. Postischemic recovery of isovolumetric developed pressure was significantly impaired in group II (16.1% +/ 7.4% [II] versus 65.5% +/- 4.8% [I], p < 0.05), and 50% of the hearts had no spontaneous cardiac activity on reperfusion. End-diastolic pressure showed significant contracture with prolonged cold perfusion: group II 57.3 +/- 13.9 mm Hg versus group I 14.8 +/- 1.8 mm Hg, p < 0.05. In the absence of ischemia, a similar relationship was observed between groups IV and III (left ventricular developed pressure 68.5% +/- 3.6% versus 82.4% +/- 4.2%, p < 0.05, and left ventricular end-diastolic pressure 23.5 +/- 6.2 mm Hg versus 13.3 +/- 2.6 mm Hg, p = not significant. Ultrastructural examination revealed severe damage to the myocardial cells and contraction band necrosis in group II (prolonged cooling and ischemia). These results suggest that prolonged cold perfusion of the nonarrested newborn heart impairs functional recovery and is therefore detrimental. When followed by a period of ischemic arrest, it further potentiates the myocardial injury and induces severe contracture. This preceding adverse effect of prolonged myocardial cold perfusion before cardiac arrest may, in part, explain the suboptimal protective effect of cardioplegia in neonates. PMID- 8412259 TI - Quantitative effects of myocardial edema on the left ventricular pressure-volume relation. Influence of cardioplegia osmolarity over two hours of ischemic arrest. AB - We previously studied edema and left ventricular pressure-volume relations in a porcine heart model in which edema occurred even with hyperosmolar crystalloid cardioplegia. This susceptibility to edema was attributed to venous occlusion and an initial 20-minute period of ischemia. Results did not demonstrate reversal of edema by hyperosmolar perfusates. Accordingly, in the present study, heart weight, myocardial water content, and left ventricular pressure-volume curves were measured before and after perfusion-induced edema in eight isolated, arrested, hypothermic porcine hearts. Cardioplegic solution was infused 2.1 +/- 0.8 minutes after the onset of ischemia, and the atrioventricular ring was not clamped during the administration of cardioplegic solution. Cardioplegic solution (1 L) was infused at intervals of 33 +/- 6 minutes at 4 degrees C. Solution osmolarity was 380 (Stanford solution) or 294 mOsm/L (Plegisol solution). The perfusion sequence was 380-1, 380-2, 294-1, 380-3. Pressure-volume relations were assessed with the use of left ventricular volume at a pressure of 10 mm Hg and the ventricular chamber stiffness constant, beta, derived from P = alpha e beta V. Perfusions 380-1 and 380-2 did not affect the pressure-volume curve. Perfusion 294-1 increased heart weight and water content (p < 0.05) and decreased left ventricular volume at 10 mm Hg compared with perfusions 380-1, 380-2, and 380-3. In addition, beta increased (0.023 +/- 0.005 versus 0.029 +/- 0.006, p < 0.05) after perfusion 294-1, compared with 380-1. Correlation coefficients for linear regressions between left ventricular volume at 10 mm Hg and heart weight and water content were r = 0.84 and r = 0.70, respectively. We conclude that under conditions similar to those used clinically, the left ventricle of the pig does not develop edema with Stanford solution (380 mOsm/L). Edema does follow Plegisol solution (294 mOsm/L) cardioplegia. Edema and reduced compliance are incompletely reversed by hypertonic cardioplegia. The porcine left ventricle can usefully replicate the clinical model. PMID- 8412260 TI - The fate of circulating megakaryocytes during cardiopulmonary bypass. AB - Megakaryocytes with intact cytoplasm normally leave the bone marrow, enter central venous blood, and are filtered in the lungs. During cardiopulmonary bypass, large megakaryocytes are not filtered by the lungs and may not be removed in the extracorporeal circuit by arterial line filters. In such circumstances, they could enter the systemic circulation and block smaller cerebral vessels, resulting in neurologic impairment. To investigate the fate of circulating megakaryocytes during cardiopulmonary bypass, central venous blood and oxygenated blood samples before and after arterial line filtration (40 microns polyester screen filter) were obtained from 10 patients undergoing cardiopulmonary bypass. Megakaryocytes were isolated by whole blood filtration and identified by their characteristic structure after May-Grunwald-Giemsa staining. In preliminary studies, megakaryocyte identification was verified by immunolabeling. All samples contained megakaryocytes with copious cytoplasm. Their frequencies in central venous blood and oxygenated blood before and after the arterial line filtration (corrected for hemodilution) were 23.4 +/- 9.3 per milliliter (mean +/- standard error of the mean, range 3.1 to 89.7 per milliliter), 21.0 +/- 8.2 per milliliter (2.0 to 84.2 per milliliter) and 17.1 +/- 7.4 per milliliter (3.1 to 80.4 per milliliter), respectively. Megakaryocytes with scant or no visible cytoplasm were also observed. The results confirm that circulating megakaryocytes are a normal physiologic component. During cardiopulmonary bypass, megakaryocytes with copious cytoplasm (mean diameter 42.7 microns, range 22 to 78 microns) can pass through the extracorporeal circuit. In the absence of filtration by the lungs, these large cells have access to the systemic circulation. This study supports a possible role for circulating megakaryocytes in the development of cerebral dysfunction after cardiopulmonary bypass. PMID- 8412261 TI - Clinical trial of cefamandole, cefazolin, and cefuroxime for antibiotic prophylaxis in cardiac operations. AB - A relatively large number of comparative trials of antibiotic prophylaxis in cardiac surgery have been published, many of which have serious design flaws. Despite the large number of studies, no single antibiotic regimen has emerged as clearly superior in preventing postoperative site infections. To determine if a superior regimen could be identified with a study designed to avoid flaws found in previous studies, we undertook a randomized, double-blind clinical trial of three cephalosporins. From March 1987 to February 1990, 2759 adults underwent median sternotomies: 1641 completed study participation, 203 were enrolled but were dropped from the study for protocol violations, and 815 were excluded. The characteristics of all 2759 patients were recorded with respect to case mix and infection risk factors, and the patients were followed-up by the same nurse throughout hospitalization and for 6 weeks after discharge for the assessment of infection outcome status. Of the 1641 participants, 141 (8.6%) had one or more operative site infections: 46 of 549 (8.4%) cefamandole recipients, 46 of 547 (8.4%) cefazolin recipients, and 49 of 545 (9.0%) cefuroxime recipients (p = 0.92). The sites of infection and the depth of tissue involvement were not significantly different across groups. Because no differences in effectiveness in preventing postoperative site infections were demonstrated in a rigorously designed trial, the costs of the drugs, including the costs of their preparation and delivery, may be the only variables by which to choose among these three antibiotic prophylaxis regimens. PMID- 8412262 TI - Recovery of cerebral blood flow and energy state in piglets after hypothermic circulatory arrest versus recovery after low-flow bypass. AB - A miniature piglet model that replicates clinical hypothermic (14 degrees C nasopharyngeal) circulatory arrest and low-flow (50 ml/kg per minute) bypass was used to study carotid blood flow with electromagnetic flow probe, cerebral blood flow by microsphere injection, cerebral metabolic rate by arteriovenous oxygen and glucose extractions, lactate production by cerebral arteriovenous difference, and cerebral edema. Data from five animals that underwent circulatory arrest and five animals that underwent low-flow bypass (aged 28.8 +/- 0.4 [mean +/- standard error of the mean] days) were analyzed. The duration of circulatory arrest and low-flow bypass was 1 hour. In a parallel study with the same animal model, phosphorus 31 magnetic resonance spectroscopy was used to assess cerebral phosphocreatine, nucleoside triphosphate (adenosine triphosphate), and intracellular pH. Five animals (aged 31.8 +/- 1.1 days) underwent circulatory arrest, and five underwent low-flow bypass. A brief phase of hyperemic carotid blood flow was seen immediately after the onset of reperfusion in the circulatory arrest group but not in the low-flow group. In the circulatory arrest and low flow bypass groups, cerebral blood flow (percentage of baseline 71.2% +/- 8.3% and 69.1% +/- 5.8%, respectively), cerebral oxygen consumption (45.6% +/- 10.0%, 44.5% +/- 7.6%), and cerebral glucose consumption (31.5% +/- 30.7%, 83.5% +/- 24.2%) remained depressed after 45 minutes of reperfusion and rewarming to normothermia. However, after 3 more hours of pulsatile normothermic reperfusion, cerebral oxygen consumption and cerebral glucose consumption had returned to baseline. Phosphocreatine, adenosine triphosphate, and pH were maintained at or above baseline levels throughout low-flow bypass and throughout 3 hours of normothermic reperfusion. In contrast, both phosphocreatine and adenosine triphosphate became undetectable 32 +/- 3.7 minutes after onset of circulatory arrest. During and early after circulatory arrest, pH decreased to a minimum of 6.506 +/- 0.129 at 40 minutes after reperfusion. After 3 hours of normothermic reperfusion, phosphocreatine and adenosine triphosphate recovered to 98.6% +/- 9.0% and 90.1% +/- 13.5% of baseline, respectively, and pH was 7.087 +/- 0.051, similar to baseline (7.1755 +/- 0.041). In the low-flow bypass group, the disparity between the depressed level of cerebral oxygen consumption and normal high-energy phosphate levels may reflect incomplete cerebral rewarming or decreased energy consumption. In the circulatory arrest group, the parallel recovery of oxygen consumption and high-energy phosphates eventually achieving baseline levels suggests that the degree of hypothermia used provides adequate protection for acute cerebral recovery after 1 hour of circulatory arrest.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412263 TI - Outpatient mediastinoscopy. AB - The reported experience with outpatient mediastinoscopy is limited. We have performed 65 mediastinoscopies in a hospital-based ambulatory surgical unit during the past 2 1/2 years. This represents 54% of our total mediastinoscopies during the period and 85% of the total for the past year. One patient was admitted overnight because of hypoxemia that was relieved by thoracentesis. All other patients were discharged from the outpatient recovery room without problems. No other early and no late complications occurred. The cost savings were substantial, and patient satisfaction was high. We conclude that mediastinoscopy can be performed safely in the outpatient setting in many patients. PMID- 8412264 TI - Talc pleurodesis. Experience with 360 patients. AB - Talc was used intrapleurally for the creation of pleural adhesions in 360 patients. The indications for use were malignant pleural effusion in 169 patients, benign effusions in 41, recurrent pneumothorax in 122, empyema in 19, and chylothorax in 9. Of 336 patients available for follow-up, excellent results (complete pleurodesis) were achieved in 284 patients (84.5%), fair results in 28 (8.3%), and poor results (no adhesions) in 24 (7.2%). The results were best in empyema (100%) and better in pneumothorax than in pleural effusions. With repeat talc insufflation, the results were improved to excellent in more than 90% of the patients. Talc is the most effective and least expensive agent for creation of pleural adhesions. Its use is simple and easily tolerated by the patients. Because of the carcinogenic effect of asbestos, purified talc that is free of asbestos fibers must be used. PMID- 8412265 TI - Multimodality therapy of patients with stage IIIA, N2 non-small-cell lung cancer. Impact of preoperative chemotherapy on resectability and downstaging. AB - To assess the effect of neoadjuvant platinum-based chemotherapy on resectability, stage of disease at resection, and patterns of recurrence and survival in patients with IIIA, N2 non-small-cell lung cancer, we examined the first 60 patients treated with neoadjuvant chemotherapy followed by attempted resection in our institution. Of 67 patients identified, 7 patients were ineligible because of comorbidities, 3 patients refused chemotherapy, and 1 consented but died before treatment. Fifty-six received neoadjuvant chemotherapy. Complications of chemotherapy were minor, with no deaths. Fifty-four patients had thoracotomy; 75% (n = 42) had complete resection and 25% (n = 14) had unresectable lesions. One postoperative death occurred (2%). Pathologic review of specimens and nodal groups revealed that 41% (n = 23) were downstaged, 39% (n = 22) remained stage IIIA, and 19% (n = 11) progressed. Squamous histologic type was predictive of resectability, 18 of 20 patients having resectable squamous cell tumors (p < 0.05). Actuarial survivals at 1 and 2 years were 74% and 52%, respectively. In patients with resectable tumors survivals at 1 and 2 years were 85% and 67%, respectively. For those with unresectable lesions, survivals were 43% and 14%. Relapse-free survivals at 1 and 2 years for patients with resectable lesions were 70% and 42%, respectively. Relapses were local in 25% (n = 4), at a distant site only in 50% (n = 8), combined local and distant in 25% (n = 4). Distant relapse occurred in the central nervous system only in 7 of 8 patients (88%). Complete resectability was highly predictive of improved survival (p < 0.0002). Weight loss did not affect resectability but was associated with decreased survival (p < 0.003). Neoadjuvant chemotherapy appears to improve resectability and to pathologically downstage N2 non-small-cell lung cancer from stage IIIA. Multiinstitutional randomized trials are needed to further demonstrate the efficacy of this approach. PMID- 8412266 TI - Adjuvant chemotherapy for completely resected stage III non-small-cell lung cancer. Results of a randomized prospective study. The Japan Clinical Oncology Group. AB - Two hundred nine patients with completely resected stage III non-small-cell lung cancer were randomized to receive postoperative cisplatin and vindesine chemotherapy or no further treatment. Before randomization, patients were stratified by the histologic characteristics of their tumors (squamous versus nonsquamous cell carcinoma). Prognostic variables such as histology, performance status, extent of operation, and tumor and nodal status of the eligible patients in chemotherapy (n = 90) and control groups (n = 91) were equally distributed. There was no statistically significant difference in disease-free and overall survival between the two groups. The 3-year disease-free survivals of the chemotherapy and control groups were 37% and 42%, respectively. The median survival times (5-year survival) were 31 months (35%) in the chemotherapy group and 37 months (41%) in the control group. These was no different pattern in the first site of recurrence (local versus systemic) between the two groups. This study failed to demonstrate the therapeutic benefits of postoperative cisplatin and vindesine chemotherapy. PMID- 8412267 TI - Prognosis of patients with hypertrophic obstructive cardiomyopathy after transaortic myectomy. Late results up to twenty-five years. AB - A complete clinical study was performed for 364 patients with hypertrophic obstructive cardiomyopathy who were operated on in the years 1963 to 1991 (217 male, 146 female, mean age 40 years, range 5 months to 76 years). Transaortic subvalvular myectomy was performed in 272 patients (hospital mortality 2.9%), and 92 patients needed additional cardiac procedures simultaneously (hospital mortality 10.9%). A complete follow-up study (100%) included 346 patients who survived the operation. The shortest follow-up time was 2 months and the longest 25.2 years (mean 8.2 years). Most of the patients improved clinically by one to three classes (New York Heart Association). During the observation period 38 patients (10.4%) died. The death of 17 patients was closely related to the original disease (4.9%). Other causes, unrelated to hypertrophic obstructive cardiomyopathy, were responsible for the death of 21 patients (5.8%). In consideration of these data, the yearly total death rate was 2.2%; in close relation to hypertrophic obstructive cardiomyopathy it was about 0.6%. The cumulative survivals were 88% after 10 years and 72% after 20 years. In our long term clinical experience it is increasingly evident, despite the restrictions of a retrospective study, that patients with symptomatic hypertrophic obstructive cardiomyopathy and failing medical therapy benefit from transthoracic subvalvular myectomy. PMID- 8412268 TI - Anomalous origin of the left coronary artery from the pulmonary artery. New operative technique. AB - We report a new operative technique for repair of anomalous origin of the left coronary artery from the pulmonary artery. The principles of the proposed technique are left main coronary angioplasty using a transected main pulmonary artery, side-to-side anastomosis of the aorta and newly created left coronary artery, and direct anastomosis of the transected pulmonary artery. No prosthetic material is used in this procedure. Our experience in two adults (a 35-year-old man and a 68-year-old woman) indicated that this technique permits two coronary system repair for any anatomic variation of the left coronary artery without the use of prosthetic material. This is more advantageous in infants. PMID- 8412269 TI - Left ventricular function assessed with echocardiography and myocardial perfusion assessed with scintigraphy under dipyridamole stress in pediatric patients after repair for anomalous origin of the left coronary artery from the pulmonary artery. AB - Twenty-three patients who underwent operation for anomalous origin of the left coronary artery from the pulmonary artery were reexamined with two-dimensional echocardiography and thallium 201 perfusion imaging. Follow-up studies were performed 0.6 to 16.2 years (median 2.9 years) after operation. In 22 of 23 patients, a two coronary artery system had been established by implantation of the left coronary artery into the aorta (n = 8) or by anastomosis of the left subclavian artery with the left coronary artery (n = 14). The left coronary artery had been ligated in only one patient. For stress testing, 0.8 mg dipyridamole per kilogram body weight was infused in a 10-minute period in 20 of the 23 patients. High-dose dipyridamole infusion increased mean heart rate (98.1 +/- 27.1 to 122.3 +/- 19.2 beats/min, p < 0.001) and mean left ventricular ejection fraction (54.8% +/- 11.8% to 61.3% +/- 12.5%, p < 0.05) and decreased left ventricular end-diastolic volume index (38.8 +/- 26.7 to 29.9 +/- 8.3 ml/m2, p < 0.005). At rest, left ventricular dimensions were abnormal in only one patient, in whom the anastomosis with the left coronary artery proved to be occluded, as seen with subsequent angiography. Left ventricular function seen with two-dimensional echocardiography was normal in 19 patients and was compromised in 3 (all of whom had major structural anomalies of the left ventricle, such as left ventricular aneurysm, occlusion of the anastomosis, or mitral valve prosthesis). Patients with R-wave loss as seen with preoperative electrocardiography tended to have larger left ventricular volumes at follow-up (69.2 +/- 56.5 ml/m2 versus 32.4 +/- 9.6 ml/m2, p < 0.07). Ten of 20 patients had normal thallium 201-perfusion scans. In 9 of 20 patients defects revealed by permanent thallium 201-perfusion were observed and determined to be myocardial scars. Transient perfusion defects under dipyridamole stress with redistribution at rest occurred in three children, two of whom also had permanent thallium 201 defects. None of the three patients had angina-like symptoms or S-T segment changes during dipyridamole stress. Left ventricular ejection fraction, however, decreased severely during dipyridamole infusion in the single patient with ligature of the left coronary artery. The two remaining patients had normal echocardiographic left ventricular function under stress, and the diagnosis of myocardial ischemia as seen with scintigraphy must be questioned.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412270 TI - Pulmonary atresia with intact ventricular septum. Surgical approach based on ventricular size and coronary anatomy. AB - Pulmonary atresia with intact ventricular septum has continued to have a high surgical mortality. This may relate to the nonuniformity of the anomaly. We have developed a management algorithm based on the right ventricular size and coronary anatomy. Patients with a well-developed ventricle and normal coronary arteries have undergone right ventricular outflow procedures. The adequacy of their right ventricles is subsequently evaluated; some patients are candidates for a four chamber repair, whereas others are candidates for a Fontan procedure. Patients with severe hypoplasia of the right ventricle frequently have extensive ventriculo-coronary connections, and for these patients we have developed the technique of right ventricular obliteration. Over the past 7 years, we have treated 20 patients with pulmonary atresia with intact ventricular septum. Fourteen of the 20 patients underwent outflow tract procedures, with no operative mortality. There were two late deaths in this group. Six patients had hypoplasia of the ventricle with ventriculo-coronary connections and underwent right ventricular obliteration. There was one operative death and one late death in the group. Overall, 95% survived the neonatal period, and 80% were still alive at the time this article was written with an average follow-up of 32 months. We conclude that pulmonary atresia with intact ventricular septum can be successfully managed with the use of an algorithm based on ventricular size and coronary anatomy. PMID- 8412271 TI - Pulmonary microthrombi. Caveat for successful modified Fontan operation. AB - We reviewed medical records from eight patients (4 to 29 years of age) with a functional single ventricle and pulmonary microthrombosis as observed on open lung biopsy specimens. Hemodynamic assessment before biopsy revealed pulmonary hypertension (mean pulmonary artery pressure 19 to 53 mm Hg) in the seven patients tested and severely increased pulmonary arteriolar resistance (6 to 13 U.m2) in three of the five patients in whom these measurements were performed. Pulmonary blood flow varied (< 4 L/min per square meter in two patients and > 5 L/min per square meter in four patients), and the pulmonary/systemic blood flow ratio was low or normal (0.6 to 1.1) in five of six cases. Hemoglobin values were increased (16.4 to 22 gm/dl) in seven of eight patients. Lung biopsy specimens revealed thrombotic obstructive lesions in 56% (range 28% to 96%) of muscular pulmonary arteries. Coexistent medial hypertrophy was absent or mild in all but one specimen, and none had features of plexogenic pulmonary arteriopathy. Microscopic pulmonary thrombi accounted for unexpected pulmonary hypertension, precluding the Fontan operation, in five patients and unsuccessful Fontan operation in two. Clinically significant thrombi should be considered in patients with severe polycythemia (hemoglobin value > 20 gm/dl) and unobtainable or unreliable pulmonary blood flow or pulmonary resistance determinations. PMID- 8412272 TI - Invited letter concerning: myocardial temperature management during aortic clamping for cardiac surgery: protection, preoccupation, and perspective (J Thorac Cardiovasc Surg 1991;102:895-903) PMID- 8412273 TI - Reoperation for excessive bleeding after cardiac operations. PMID- 8412274 TI - Noninvasive preoperative and postoperative serial hemodynamic assessment of the internal mammary artery in myocardial revascularization. PMID- 8412275 TI - Facilitated internal mammary artery anastomoses. PMID- 8412276 TI - Management of cold agglutinemia with warm heart surgical intervention: a case report. PMID- 8412277 TI - Transesophageal echocardiography: an increasing role in the diagnosis of traumatic aortic rupture. PMID- 8412278 TI - Paravertebral extramedullary hematopoiesis (as a posterior mediastinal tumor) associated with congenital dyserythropoietic anemia. PMID- 8412279 TI - Invited letter concerning: normocalcemic blood or crystalloid cardioplegia provides superior neonatal myocardial protection over low-calcium cardioplegia (J Thorac Cardiovasc Surg 1993;105:201-6) PMID- 8412280 TI - Esophageal adenocarcinoma eighteen years after esophageal resection with jejunal interposition for benign disease. PMID- 8412281 TI - 81 cases of paralytic strabismus treated with acupuncture. PMID- 8412282 TI - Acupuncture treatment of chronic rhinitis in 75 cases. PMID- 8412283 TI - Treatment of acute lumbar sprain with otoacupuncture at the acupoint lumbago. PMID- 8412284 TI - Clinical and experimental studies on shallow needling technique for treating childhood diarrhea. AB - Treatment of diarrhea in children by shallow needling and by drugs was studied in 3 separate groups for comparison. For Group I, the lateral line II of the forehead was taken, i.e. the upper, middle and lower points of the line staring from Toulinqi (UB 15) vertically downwards to the place 1 cm below the hairline were selected. For Group II, shallow needling was done on body acupoints, the main points selected being Qihai (Ren 6), Shuifen (Ren 9), bilateral Tianshu (St 25), and bilateral Zusanli (St 36); the adjuvant acupoints were Taibai (Sp 3) and Gongsun (Sp 4). The depth of shallow needling was merely puncturing the skin. For Group III the drugs used were Liteling, Berberine, Gentamycin, Ampicillin, and SMZ Co. The results of treatment in Group I and II differed insignificantly, while both were significantly superior to drugs in Group III. Experimental studies also showed that shallow needling enhanced the humoral and cellular immunity and promoted the intestinal peristaltic function. PMID- 8412285 TI - Clinical studies on the mechanism for acupuncture stimulation of ovulation. AB - Ovulatory dysfunction is commonly seen in gynecology clinic. It may cause infertility, amenia, functional uterine bleeding and a variety of complications. This research according to TCM theory records treating with acupuncture 34 patients suffering from ovulatory dysfunction. Changes in clinical symptoms and some relative targets are reported, plus findings in animal experiments. The effect of acupuncture in improving ovulation and the rationale are discussed. According to TCM theory concerning the generative and physiologic axis of women, this research involved the following points: Ganshu (UB 18), Shenshu (UB 23), Guanyuan (Ren 4), Zhongji (Ren 3), and Sanyinjiao (Sp 6). The reinforcement and reduction of acupuncture enables it to strengthen liver and kidney. Through the Chong and Ren channels it nourishes uterus to adjust the patient's axis function and recover ovulation. Treated on an average of 30 times, the patients' symptoms improved to varying degrees. The marked effective rate was 35.29%, the total effective rate being 82.35%. BBT, VS, CMS, and B ultrasonic picture all improved to some degree. The results also showed that acupuncture may adjust FSH, LH, and E2 in two directions and raise the progesterone level, bringing them to normal. The animal experiments confirmed this result. Results showed that acupuncture may adjust endocrine function of the generative and physiologic axis of women, thus stimulating ovulation. The results of this research will provide some scientific basis for treating and further studying this disorder. PMID- 8412286 TI - An introduction to keeping-fit massage (1). PMID- 8412287 TI - A survey of the treatment of obesity by traditional Chinese medicine. PMID- 8412288 TI - Recent studies on auriculoacupuncture and its mechanism. AB - This article is based on the review of more than eighty references involving clinical treatment, analgesia, auriculoacupoint for diagnosis and its mechanism. In the first part, selection of acupoints, methods of manipulation, therapeutic effects and related experience in 56 kinds of respiratory, circulatory, digestive, urological, gynaecological, obstetrical, pediatric, otolaryngologic and ophthalmologic, dermatologic diseases and diseases of the nervous systems, as well as treatment of acute abdominal diseases, reduction of body weight, abstinence from smoking and drinking, etc. are discussed. A variety of stimulation methods, such as auriculoacupuncture with filform needles, needle embedding therapy, point injection, bloodletting, laser irradiation, aurioular plaster and pressing therapy, etc are also introduced. In the second part, experiences of surgical operation and endoscopy under auriculoacupuncture anesthesia are described. The third part deals with diagnosis using auriculoacupoints. Also described in this part is the clinical application of auricular acupoints in the diagnosis of cancers, coronary diseases, cholelithiasis, hepatitis, tertian malaria, etc. The correlation between the morphology and function of auricular acupoints, i.e. the relationship between auricular acupoints and visceral function, and main development in clinical therapy, diagnosis and mechanism of auriculoacupuncture are introduced in the fourth part, including the results of investigations of reactions of auriculoacupoints during disorders of the body and viscerae, reactions of viscerae during stimulation of auriculoacupoints and their transmission routes. Based on the analysis of the history and current status of research on auriculoacupuncture, the author emphasizes the necessity of attaching importance at the same time to prophylaxis and treatment of serious diseases and standardization of nomenclature. Suggestions in this connexion are also made. PMID- 8412289 TI - Health preservation by mental means (5). PMID- 8412290 TI - Clinical application of traditional auriculoacupoint therapy. PMID- 8412291 TI - Treatment of diabetes in the elderly--an analysis of 885 cases. AB - Besides treating elderly diabetics basically for the disease, the author treated the patients by traditional Chinese medicine based on an overall analysis of their symptoms and signs, dividing the course of the disease into 5 stages comprising 16 syndromes types. He evaluated the quality of their survival according to an the health rating of Karnofsky, and advised them on the amount of work or other activity appropriate in each case in order to relieve the symptoms, retard the disease process, and improve the quality of survival. PMID- 8412292 TI - Treatment of AIDS with traditional Chinese medicine. PMID- 8412293 TI - 215 cases of obesity treated with TCM modalities. PMID- 8412294 TI - Age and Hodgkin's disease: the impact of competing risks and possibly salvage therapy on long term survival: an E.C.O.G. study. AB - A detailed review of factors associated with survival was carried out in a cohort of 560 patients treated in two successive E.C.O.G. studies on advanced Hodgkin's disease. The study was undertaken to explore the impact of age on survival and to attempt to identify reasons for any observed differences. Data from two E.C.O.G. studies of patients with advanced Hodgkin's disease were examined separately and then pooled together. A special data request form was developed to capture additional information on treatments utilized for patients who were treated at relapse. The complete remission percentages were identical in both studies (72%) with no significant difference between the three age groupings (< 40, 40-59, and > 60 yr). This was true as well for disease-free survival. Nevertheless, overall survival was significantly better for the under aged 40 group and this difference was narrowed but not eliminated by competing risks. Our analysis of salvage therapy revealed a marginally significant difference in the CR% between the three groups, favoring the youngest cohort (< age 40 yr). The identical remission rate is probably a reflection of the entry criteria eliminating poor risk patients. Among the small number of patients who received radiation therapy, the response rate was 75% (9/12) in the young cohort. Elderly patients appeared to receive less salvage therapy and certainly fewer responses. Survival after relapse showed a similar pattern with the best survival in the youngest group of patients followed closely by the 40-59 yr group, with a poor outcome for those over age 60 yr. We conclude that a variety of conditions, both age and non-age dependent, impact on the survival of patients with advanced Hodgkin's disease who fulfill the rigid criteria for entry into cooperative group trials. PMID- 8412295 TI - Chronic myeloid leukemia granulocytes have lower amounts of cytoplasmic actin. AB - Chronic myeloid leukemia (CML) granulocytes had earlier been shown to be defective in microfilament mediated phenomena such as chemotaxis and fluid phase pinocytosis when compared to normal granulocytes. The present studies were carried out to determine whether a quantitative alteration in actin or a change in isoform status could be responsible for some of the defects. Relative proportions of beta and gamma isoforms were found to be unaltered between normal and CML granulocytes. The amount of actin was significantly lower in CML cells. When the actin was expressed as percent of total protein in the granulocytes, it was found to be significantly lower in CML cells. The lower amount of actin may be responsible for some of the defects seen in CML cells. PMID- 8412296 TI - A quantitative evaluation of erythropoiesis in myelodysplastic syndromes using multiparameter flow cytometry. AB - By staining human bone marrow cells with a monoclonal antibody reacting with erythroid precursor cells (AS-E1) and propidium iodide, we have evaluated the proliferative capacity of erythropoiesis in patients with myelodysplastic syndromes (MDS) using flow cytometry. Comparing 36 patients (13 RA/RAS, 13 RAEB, 10 RAEB-t) with 7 normal controls, significant differences in both the percentage of AS-E1+ cells and the fraction of AS-E1+ cells in the S or S-G2M-phase between the four groups were found. Since neither the percentage of AS-E1+ cells nor their fraction in S or S-G2M alone was found to characterize their proliferative activity, we introduced the proliferative fractions of the erythroid cell, i.e. the number of the AS-E1+ cells in S or S-G2M related to all bone marrow cells in S or S-G2M. Applying these parameters, we found significantly increased proliferative AS-E1 fractions in the RA/RAS group compared to the normal controls (p = 0.03 and 0.002) respectively, as well as a highly significant decrease with disease progression. PMID- 8412297 TI - Genistein exhibits preferential cytotoxicity to a leukemogenic variant but induces differentiation of a non-leukemogenic variant of the mouse monocytic leukemia Mm cell line. AB - Mouse leukemia Mm-A and Mm-S2 cells are subclones of mouse monocytic leukemia Mm cells, Mm-A cells having much higher leukemogenicity than Mm-S2 cells. The growth inhibitory effects of several protein kinase inhibitors on leukemogenic Mm-A and non-leukemogenic Mm-S2 cells were examined. Most inhibitors of protein serine/threonine kinases inhibited the growth of Mm-A and Mm-S2 cells similarly, but some protein tyrosine kinase inhibitors exhibited differential inhibitory effects on Mm-A and Mm-S2 cells. Genistein inhibited growth of Mm-A cells more effectively than that of Mm-S2 cells, but another inhibitor of tyrosine kinase, herbimycin A, preferentially inhibited growth of non-leukemogenic Mm-S2 cells. Genistein induced or enhanced several differentiation markers of Mm-S2 cells, such as cell spreading, immunophagocytosis, nitroblue tetrazolium (NBT) reduction and lysozyme activity in a dose-dependent manner, but herbimycin A did not. Genistein was cytotoxic to Mm-A cells rather than inducing cell differentiation. Genistein has effects on several other cellular events as well as inhibition of tyrosine kinases. However, it effectively inhibited protein tyrosine phosphorylation in Mm-A cells and its decrease of tyrosine phosphorylation was closely associated with its inhibition of cell growth. Thus, a genistein sensitive tyrosine kinase(s) may play an important role in the growth and/or survival of leukemogenic Mm-A cells. PMID- 8412298 TI - Iron responsive element-binding protein (IRE-BP) in leukemic cells: analysis using enzyme-linked immunosorbent assay and semiquantitative polymerase chain reaction. AB - The level of iron responsive element-binding protein (IRE-BP) in leukemic cells, which is essential for iron homeostasis and plays an important role in cell metabolism and cell growth, was measured using an enzyme-linked immunosorbent assay (ELISA). Comparing the levels in different clinical stages, the levels in CML cells in the chronic phase (mean +/- S.E., 0.270 +/- 0.110 U/mg protein, n = 9) and those in AML cells (0.150 +/- 0.104 U/mg) protein, n = 21) were significantly lower than that in normal granulocytes (0.628 +/- 0.216 U/mg protein, n = 9, p < 0.001 vs CML and AML). Analysis of IRE-BP mRNA expression in leukemic cells using semiquantitative polymerase chain reaction showed suppressed expression of mRNA as compared to normal bone marrow cells. These observations suggest that there may be dysregulation of IRE-BP expression and production in leukemic cells. PMID- 8412299 TI - A quantitative ultrastructural and cytochemical study of TPA-induced differentiation in HL-60 cells. AB - The effects of the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate on morphometric and stereological parameters have been studied using the HL-60 cell line as a differentiation model for the monocytic pathway. Evaluation of the differentiation was carried out by quantification of endoplasmic reticulum, Golgi apparatus, mitochondria and cytoplasmic granules. Changes in both nuclear and cytoplasmic volumes during TPA-induced differentiation led to a decrease of the nucleus-cytoplasmic ratio after 3 days of treatment. Plasma membrane glycoprotein pattern was also determined. The major change in cell surface was the presence of high amounts of glycoproteins containing N-acetyl glucosamine residues that make wheatgerm agglutinin lectin a valuable marker of the monocytic differentiation pathway in HL-60 cells. PMID- 8412300 TI - Establishment and characterization of the tumors of chronic lymphocytic leukemia cell line in nude and SCID mice. AB - A new cell line, designated MO1043, was established from the peripheral blood (PB) of a patient with B-cell chronic lymphocytic leukemia (CLL). Both the PB leukemia cells and MO1043 were found to have an abnormal cytogenetic marker of trisomy 12, the most common cytogenetic abnormality in CLL. In addition, both the PB cells and MO1043 expressed a cell surface phenotype of typical B-CLLs. The MO1043 was efficiently transplanted into X-irradiated athymic nude mice by i.p. inoculation after it was subjected to serial passages in new born (1 week old) and irradiated adult nude mice. The tumor of a CLL cell line (termed CLL tumor) was also generated in the nude mice by s.c. inoculation of the cells. The MO1043 was inoculated i.p. into mice with severe combined immunodeficiency (SCID) which had not been subject to any preconditionings. The CLL tumor in the non conditioned SCID mice was disseminated to various tissues in a manner more analogous to CLL tumors in patients as compared with nude mice, where the CLL tumors were not as widely disseminated. At each of four different tumor doses, i.e. 2 x 10(6), 6 x 10(6), 1.8 x 10(7) and 5.4 +/- 10(7) cells of MO1043, the transplantability was 100%. Titration experiments revealed a reciprocal relationship between survival and the number of tumor cells inoculated. FACS analysis showed that several cell surface markers of the parental MO1043 were maintained in CLL tumors from nude and SCID mice. Fluorescence in situ hybridization with novel DNA probes demonstrated that CLL tumors of both nude and SCID mice maintained trisomy 12. The CLL tumor models developed here, particularly the SCID mouse model, may be very useful for therapeutic studies of CLL. PMID- 8412301 TI - Different effects of the two protein kinase C activators bryostatin-1 and TPA on growth and differentiation of human monocytic leukemia cell lines. AB - The effects of bryostatin-1 (Bryo) and 12-O-tetradecanoyl-phorbol 13-acetate (TPA), both activators of protein kinase C (PKC), on proliferation and differentiation of two monocytic leukemia cell lines, JOSK-I and JOSK-M, were investigated. Treatment with TPA or Bryo inhibited cellular proliferation in a dose-dependent manner. Both drugs induced distinct phenotypic changes associated with monocytic differentiation. Although c-myc mRNA is often found to be down regulated during biomodulator-triggered in vitro myelomonocytic differentiation, however, here the modulation of c-myc expression was less pronounced. All parameters studied were more prominently altered in TPA- than in Bryo-treated cells, and, were more distinct in JOSK-I than in JOSK-M. Since Bryo was able to antagonize the TPA-mediated effects on proliferation and morphological alterations, an (at least partially) different mode of action of these PKC activators on monocytic cell lines may be suggested. PMID- 8412302 TI - Overview of chemical sampling techniques. AB - Although many of the ideas for sampling the chemical microenvironment of the brain were present, at least in nascent form, three decades ago or more, the last 10 years have witnessed a particularly spectacular surge of development, refinement, and use. We are now able to measure virtually any endogenous brain chemical in vivo at commendable levels of sensitivity, selectivity, and speed. The long-dreamt-of goal of being able to correlate neurochemical events with ongoing behavior and/or presentation of salient environmental cues and stimuli has already been largely achieved. Further refinements of existing techniques may well lead to levels of analysis inconceivable even a few years ago. The implications for theory-building and hypothesis-testing are enormous, particularly within such essentially virgin domains as behavioral neuroscience and biological psychiatry. These are truly exciting times. PMID- 8412303 TI - Ion-selective microelectrodes and diffusion measurements as tools to explore the brain cell microenvironment. AB - The construction and application of liquid-membrane ion-selective microelectrodes (ISM) are described. Recommendations are provided for the selection of appropriate cocktails containing neutral carriers to form the liquid membrane to sense K+, Ca2+, H+ and Na+. The use of charged carriers to sense Cl- and the cation tetramethylammonium (TMA+) is discussed. A detailed protocol is given for constructing double-barreled electrodes (ion-sensor and reference barrel) with tips of 1 micron diameter or more for extracellular ion measurements. The primary results obtained with ISMs in the brain cell microenvironment are briefly surveyed. The theoretical basis for measuring diffusion properties of extracellular space is described. Such measurements enable the estimation of volume fraction (proportion of tissue that is extracellular space) and tortuosity (hindrance of diffusion due to cellular obstructions). A method is given for using TMA+ ISMs in combination with iontophoresis or pressure ejection of TMA+ from a nearby micropipette to measure diffusion properties. PMID- 8412304 TI - Microcolumn separations of single nerve cell components. AB - Miniaturization of separation techniques is providing new methods to examine neurochemistry and neurophysiology at the single cell level. Open tubular liquid chromatography and capillary electrophoresis have been scaled down to levels where picoliter and femtoliter volume samples can be separated. This provides the means to profile the chemistry of single whole nerve cells and to sample subcellular regions. PMID- 8412305 TI - Principles of voltammetry and microelectrode surface states. AB - In vivo voltammetry is approaching the end of its second decade as a technique to explore extracellular concentrations in the brain. The issues of selectivity and sensitivity, which caused considerable discussion and confusion in the early 1980s, are now resolved. It is clear that in vivo voltammetry and dialysis are complimentary tools to understand neurotransmitter dynamics. The two chief advantages of voltammetry compared to dialysis, improved temporal resolution and reduced tissue damage, make this technique exceptionally well suited for providing information which is complementary to that obtained by single-unit recording and is uniquely capable of providing information on the short-term regulation of extracellular levels of biogenic amines. PMID- 8412306 TI - High sensitivity measurement of brain catechols and indoles in vivo using electrochemically treated carbon-fiber electrodes. AB - The combination of electrochemically treated carbon-fiber electrodes with DPV, DNPV or DPA represents a wide range of possibilities. As shown in this review, the choice of treatment and measurement technique depends on the purpose. As regards in vivo monitoring of 5-HIAA or DOPAC from very small brain nuclei, electrochemically treated carbon-fiber electrodes appear very potent and inexpensive. The main limitation of the established electrochemical techniques, including those discussed here, is that the unequivocal measurement of the basal extracellular neurotransmitter level cannot be achieved unless animals are treated with pargyline. On the other hand, this monitoring is feasible with in vivo dialysis. Therefore, electrochemical techniques, on the one hand, and in vivo dialysis, on the other hand, present different advantages. The former are much more potent than the latter in two respects. First, due to the much smaller size of the sensor, electrochemical techniques are more suitable for studying small brain nuclei. Second, since electrochemical techniques exhibit a better temporal resolution, they are recommended for investigating the relationship between impulse flow and neurotransmitter release. However, when high anatomical or temporal resolution is not required, in vivo dialysis is more suitable for recording the basal monoamine release. PMID- 8412307 TI - Respiratory symptoms, lung function tests, airway responsiveness, and bronchoalveolar lymphocyte subsets in B-chronic lymphocytic leukemia. AB - A respiratory questionnaire was completed and spirometry, tests for lung volumes, diffusion capacity for CO, and methacholine bronchial challenge were performed in 24 outpatients with B-chronic lymphocytic leukemia (B-CLL), aged 44-79, presenting in different stages of their disease. In 10 patients, bronchoalveolar lavage (BAL) fluid was also obtained. Ten of twenty-four patients had symptoms consistent with chronic bronchitis, unrelated both to smoking history and to the clinical stage. Abnormal values (< 2 SD) were found in 4 patients for total lung capacity (TLC), in 9 for vital capacity (VC), 8 for forced expiratory volume in 1 sec (FEV1), 11 for MEF50, 15 for MEF25 and in 7 for diffusing capacity for carbon monoxide. Seven of nineteen patients had PD20FEV1 at less than 1,600 micrograms of methacholine chloride. There was a significantly negative correlation between white blood cell count and VC (r = 0.41, P < 0.05). A positive correlation was found between PD20FEV1 and FEV1/VC (r = 0.61, P < 0.01). The mean and SEM for BAL cells/ml was 463 (71.8) x 10(3). No leukemic cells but a marked increase in T lymphocytes (32.5 +/- 7.8%) were found in BAL fluid. There were significantly negative correlations between the number of BAL CD3+ T lymphocytes and PD20FEV1 (r = 0.61, P < 0.05), and between the number of BAL CD8+ T lymphocytes and PD20FEV1 (r = 0.84, P < 0.01). In conclusion, patients with B-CLL have a high prevalence of respiratory symptoms, small airway dysfunction and CD8 "alveolitis" related to airway responsiveness; despite the well-known lung interstitial lymphocyte infiltration in B-CLL, leukemic cells are not found in BAL fluid. PMID- 8412308 TI - Early effects of short-time cigarette smoking on the human lung: a study of bronchoalveolar lavage fluids. AB - We investigated the early effects of cigarette smoking in healthy subjects by means of lung lavage, looking at markers of alveolar permeability, the alveolar cell profile, the immunophenotyping of macrophages and lymphocytes, and the level and profile of surfactant phospholipids. Bronchoalveolar lavages (BAL) were performed in 33 healthy subjects [20 nonsmokers (nS), 13 moderate and short-time smokers (S)]. In the acellular supernatants we measured the markers of alveolar permeability (i.e., total proteins, albumin, albumin/urea), the alveolar epithelial lining fluid (AELF), the surfactant amounts and profile, and explored the blood lymphocytes by in vitro exposure. The cell pellet established the alveolar formula and a membrane mapping of macrophages (LFA-1 and HLA-DRII expression) and lymphocytes (CD4, CD8, LFA-1, HLA-DRII expression). We found no significant increase of alveolar permeability in our smokers, but an increased alveolar cellularity (more than 3-fold vs nS, P < 0.05) evenly distributed between sub-populations except for an enhanced number of eosinophils in smokers (P < 0.05 vs nS). Smokers' alveolar macrophages had an overloaded cytoplasm, a decreased percentage of antigen-handling cell expression (HLA DRII: P < 0.05 vs nS) and a low percentage of cell to cell adhesion molecule expression (LFA-1: P < 0.05 vs nS). Smoking history and LFA-1 expression on alveolar macrophages were interrelated. Smokers' alveolar lymphocyte subsets were more often T suppressor cells (CD8+) and had an increased percentage of antigen-presenting cell expression (HLA DRII: P < 0.05 vs nS). Smokers' BAL fluid did not show the inhibitory control of phytohemagglutinin-induced lymphocyte proliferation present in nonsmokers' fluids. Surfactant phospholipid amounts were similar, but phosphatidylethanolamine was raised and the ratio of phosphatidylcholine to sphingomyelin decreased in smokers (P < 0.05 vs nS). We observed specific cellular and biochemical alterations in the lung lavage of short-time smokers. Alveolar macrophage and lymphocyte expression of LFA-1 and HLA-DR II molecules was altered. Smokers' alveolar fluids lost the physiologic regulatory control of T mitogen-induced lymphocyte proliferation. Membrane phospholipids released by cellular damage increased early in tobacco-exposed lung fluids. This profile of alterations may be an early and sensitive marker of smoking-induced lung damage. PMID- 8412309 TI - Mitoxantrone and cytosine arabinoside as treatment for acute myelogenous leukemia (AML) at first recurrence. AB - A total of 107 patients with newly diagnosed acute myeloblastic leukemia (AML) were referred to the ICRF Department of Medical Oncology at St Bartholomew's Hospital between August 1986 and July 1989. Of those referred, 92 (87%) were treated with remission induction chemotherapy comprising: Adriamycin, cytosine arabinoside (ara-C) and 6-thioguanine if aged < 60 years (57 patients) or mitoxantrone (MTN) and ara-C if aged > 60 years (35 patients). Of those treated, 54 (58%) entered complete remission (CR). Recurrent AML developed in 38 out of these 54 patients (70%) of whom 25 aged 19-73 years (median 50 years) subsequently received MTN and ara-C as reinduction therapy. The 19 younger patients (under 60 years old) received MTN at 12 mg/m2, intravenously, daily for 5 days and ara-C at 100 mg/m2, intravenously, twice daily for 7 days. The six older patients received the same ara-C schedule but the dose of MTN was reduced to 10 mg/m2 for 5 days. Second CR was achieved in 16 out of 25 patients (64%) [12/19 (63%) and 4/6 (67%) for patients aged under or over 60 years, respectively]. Eight of the patients in whom second CR was achieved were aged under 50 years and were thus eligible for additional consolidation comprising myeloablative therapy with autologous bone marrow transplantation (ABMT). Four patients actually received the latter treatment: two remain in second CR at 21 and 46 months. Three of the remaining eight patients aged > 50 years in whom second CR was achieved remain in second CR 8 to 43 months later. Censored for myeloablative therapy + ABMT, the overall median duration of second CR was 5 months. Although remissions tended to be short, in younger patients the possibility of proceeding to myeloablative therapy with autologous bone marrow support makes the regimen worthwhile and, even in older patients, it was sometimes possible to achieve prolonged second remissions. PMID- 8412310 TI - A phase I trial of carboplatin and etoposide given as continuous infusions to adults with leukemia. AB - Thirty-two adult patients with acute leukemia were treated with continuous infusion carboplatin and etoposide at eight different dose levels. Dose limiting toxicity was encountered at doses of carboplatin of 310 mg/m2 per day for 5 days (total 1550 mg/m2 per course) and etoposide 150 mg/m2 per day for 5 days (total dose 750 mg/m2 per course) and consisted primarily of gastrointestinal toxicity. Of the five courses given at the highest dose level, three were complicated by grade 4 gastrointestinal toxicity. The recommended dose for further phase II evaluations in acute leukemia is carboplatin 250 mg/m2 per day for 5 days by continuous infusion and etoposide 150 mg/m2 per day for 5 days given by continuous infusion. The response to treatment was notable for a single complete remission achieved in a patient with acute undifferentiated leukemia. None of the patients with chronic myeloid leukemia in blast crisis (n = 12) or with acute lymphocytic leukemia (n = 4) or acute myeloid leukemia (AML) (n = 15) achieved complete remission (CR), although all patients with AML entered on this trial had prior complete remissions that were less than 1 year in duration, and many were refractory, either to primary therapy or to the most recent attempt to obtain CR. PMID- 8412311 TI - Detection and significance of bcr-abl mRNA transcripts and fusion proteins in Philadelphia-positive adult acute lymphoblastic leukemia. AB - Blast cells from an unselected consecutive series of 84 adults presenting with acute lymphoblastic leukemia (ALL) to St Bartholomew's Hospital over a seven year period were tested prospectively by cytogenetic and retrospectively by RT-PCR analysis for the presence of the Ph translocation and bcr-abl mRNA. This combination gave an overall figure of 20.3% for bcr-abl-positive and/or Ph positive ALL. The incidence of bcr-abl-positive/Ph-positive ALL was most common between the ages of 31 and 50 years, becoming less common after the age of 50. Eight out of ten bcr-abl-positive patients expressed the e1a2 mRNA transcript, the other two expressed the b3a2 and b2a3 transcripts respectively. Cells from all patients with bcr-abl mRNA transcripts expressed the appropriate p190 or p210 bcr-abl protein and all were Ph-positive. PMID- 8412312 TI - Characterization of translocation t(1;14)(p32;q11) in a T and in a B acute leukemia. AB - The TAL1 locus on chromosome band 1p32 is rearranged in 15 to 29% of human T-cell acute lymphoblastic leukemias (T-ALLs). These alterations consist of either a tald submicroscopic deletion (12-26% of T-ALL) or a t(1;14)(p32;q11) chromosomal translocation (3% of childhood T-ALL). Both types of alterations preferentially affect the 5' part of the TAL1 locus. Their main consequence appears to be transcriptional activation of the TAL1 gene. We have characterized two cases of t(1;14)(p32;q11) in ALL. Both affect the TCR delta gene segments at 14q11 and the 5' part of the TAL1 locus at 1p32. The first case represented a 'classical' t(1;14), associated with T-ALL. Its analysis indicates the use of a recombination signal-like sequence localized in the third exon of TAL1 in the translocation process. In the other case, the rearrangement to the D delta region occurred 5' to the TAL1 transcription start sites. This case exhibited a B-lymphoid immunophenotype thus suggesting that the putative oncogenicity of TAL1 activation is not restricted to T-cell malignancies. PMID- 8412313 TI - Prognostic significance of detection of monoclonality in remission marrow in acute lymphoblastic leukemia in childhood. Australian and New Zealand Children's Cancer Study Group. AB - Techniques based on the polymerase chain reaction (PCR) to detect rearrangement of the immunoglobulin or T-cell receptor genes can detect residual disease in leukemia and hence have the potential to improve prognosis and treatment. Such techniques may involve either detection of monoclonality, which is simple and quick but has limited sensitivity, or specific detection of the leukaemic clone, which is complex and time-consuming but has high sensitivity. The PCR was used to detect monoclonal rearrangements of the immunoglobulin heavy chain and/or T-cell receptor gamma chain genes in archival marrow specimens from 185 children with acute lymphoblastic leukemia who achieved remission during two consecutive Australasian trials of treatment. A monoclonal rearrangement was detected at diagnosis in 152 (84%) patients and in these patients detection of the same rearrangement in the remission marrow at the end of induction therapy was highly significantly correlated with outcome. There were nine patients in whom polymerase chain reaction showed only the monoclonal rearrangement and eight (89%) relapsed; there were 26 patients in whom PCR showed the leukemic monoclonal rearrangement as well as polyclonal rearrangements from normal lymphocytes and 12 (46%) relapsed; and there were 117 patients in whom only polyclonal rearrangements could be detected and only 29 (25%) relapsed. In patients who relapsed, remissions were shorter in those patients in whom the leukemic rearrangements had been detected in the remission marrow. Treatment in the later trial was more intensive than in the earlier trial, the results were better and the PCR detected the leukemic rearrangement in the remission marrow in significantly fewer patients. We conclude that detection by PCR of the monoclonal gene rearrangement of the leukemic clone in remission marrow indicates that numerous leukemic cells have survived induction therapy and is a good predictor of relapse. However, due to limited sensitivity of the test, failure to detect the leukemic clone by PCR is not a sufficiently good predictor of ultimate cure. PMID- 8412314 TI - Transient myeloproliferative disorder and acute myeloid leukemia: study of six neonatal cases with long-term follow-up. AB - Six neonates with hematological and clinical pictures indistinguishable from acute myeloid leukemia were studied. Two patients had Down syndrome and three others had either +21 or i(21q) chromosomal abnormalities in their blood cells at presentation. Granulocyte-macrophage colony-forming unit assays performed in bone marrow and peripheral blood mononuclear cells revealed abnormal growth patterns in two patients; both died of progressive disease of acute myeloid leukemia. All the other four neonates with normal in vitro cell growth pattern had spontaneous remission within 7 months. Of these four patients, one remains well and in remission for 8 years and the other three developed acute myeloid leukemia at the ages of 15, 32 and 19 months, respectively. We conclude that the in vitro cell growth pattern is helpful to distinguish transient myeloproliferative disorder from congenital acute myeloid leukemia and that patients with the former condition are at risk to develop acute myeloid leukemia subsequently. PMID- 8412315 TI - Expression of oncoprotein 18 in human leukemias and lymphomas. AB - Oncoprotein 18 (Op18) is an 18 kDa intracellular phosphoprotein that appears to be up-regulated in certain neoplastic cells. In the present report we have analysed the expression of Op18 in samples of human hematopoietic disorders, mainly leukemias and lymphomas. For this purpose we have developed reagents allowing quantitative Western-blot analysis, and quantification of Op18 on the single cell level by flow cytometric analysis. The data demonstrates that a significant fraction of all lymphoma and leukemia cases express Op18 at levels that are several-fold higher than the Op18 levels ever found in benign proliferating tissue, such as bone marrows in remission and reactive lymph nodes. Thus, the results establish on the single cell level that Op18 is frequently expressed at abnormal levels in lymphoid and myeloid malignancies. Flow cytometric analysis revealed a striking qualitative and quantitative heterogeneity in Op18 expression between patients with lymphoma or leukemia. Moreover, our analysis demonstrates that the abnormal cellular levels of Op18 expression frequently found in high grade lymphoma and acute leukemia samples do not correlate with an increased fraction of cells in S phase. Finally, we also present an example of dramatic changes in Op18 expression pattern in bone marrow cells during the progression of a chemo-resistant acute leukemia. PMID- 8412316 TI - p53 gene mutations and loss of a chromosome 17p in Philadelphia chromosome (Ph1) positive acute leukemia. AB - We screened 23 cases of Philadelphia chromosome (Ph1)-positive acute leukemia (Ph1AL) for loss of a chromosome 17p and mutations in exons 2 to 11 of the p53 gene by single-strand conformation polymorphism (SSCP) analysis and DNA sequencing. Loss of a distal part of chromosome 17p including loss of a whole chromosome 17 emerged in three cases, among which two were Ph1-positive acute lymphoblastic leukemia (Ph1ALL) with point mutations within the highly conserved region of the p53 gene. Another case of Ph1-positive acute myelogenous leukemia (Ph1AML) also exhibited a p53 point mutation in company with loss of normal p53 allele, although showing normal chromosome 17 homologues. We also performed Southern blot hybridization analysis to examine p53 gene rearrangements in 13 cases of Ph1AL. We found a rearrangement in one case of Ph1ALL and a loss of heterozygosity (LOH) at the p53 locus without any rearrangement in another Ph1ALL. Both cases showed no abnormality within the entire coding region by SSCP analysis. Thus, p53 gene alterations were commonly involved in Ph1AL with loss of a 17p (two point mutations in three cases), while rarely in cases with normal chromosome 17s (one point mutation in 20 cases and one rearrangement in 13 cases). Rare p53 gene alterations in Ph1AL may therefore be related to low incidence of loss of a chromosome 17p. PMID- 8412317 TI - IL-8 mRNA expression and IL-8 production by acute myeloid leukemia cells. AB - Purified leukemic cells from 30 acute myeloid leukemia (AML) cases at diagnosis were investigated for the presence of interleukin 8 (IL-8) mRNA by Northern blot analysis. IL-8 specific transcripts were detected in uncultured blasts in 14/30 cases, 10/14 from patients with M4-M5 and 4/14 from cases with M0-M3 morphology. The transcript expression was associated with the detection of IL-8 molecule in blast cells by immunostaining performed on cytospin preparations. After 24-hour culture, a strong up-regulation or the appearance in cases negative before culture of IL-8 mRNA was observed in all cases tested, and culture supernatants contained high amounts of IL-8. Our data demonstrate that leukemic cells in AML are equipped with the functional apparatus for IL-8 production. Since IL-8 displays a wide range of biological activities, including the regulation of some membrane molecules relevant to adhesion and migration processes, its production by AML blasts might be of relevance for the pattern of leukemic growth. PMID- 8412318 TI - TNF alpha acts in synergy with GM-CSF to induce proliferation of acute myeloid leukemia cells by up-regulating the GM-CSF receptor and GM-CSF gene expression. AB - Acute myeloid leukemia (AML) cells are dependent for their survival and proliferation on hematopoietic growth factors. As tumor necrosis factor alpha (TNF alpha) can increase the proliferation of primary cultures of AML cells, we have investigated the effect of TNF alpha on the autocrine and/or paracrine growth control by one of the major AML growth factor, granulocyte-macrophage colony-stimulating factor (GM-CSF). First, a panel of AML cells were analysed with respect to their proliferative response to TNF alpha. We provide experimental evidence that TNF alpha induces both GM-CSF gene expression and up regulation of high-affinity GM-CSF membrane receptor in TNF alpha-responsive cells. This effect is not restricted to the malignant phenotype, although it could account for the selective growth advantage of the leukemic clone over the normal cells upon TNF alpha stimulation. PMID- 8412319 TI - Functional characterization of fibroblastic cells in long-term marrow cultures from patients with acute myelogenous leukemia. AB - We have investigated the role of acute myelogenous leukemia (AML) marrow-derived fibroblastic cells in the development of the hemopoietic microenvironment in vitro, and compared their functional integrity with that of their normal marrow derived counterparts. Our results indicate that the development of stromal adherent layers in AML long-term marrow culture (LTMC) depends to a large extent on the presence of fibroblastic progenitors (fibroblast colony-forming units, CFU F) in the bone marrow inoculum. Positive correlations were observed between the numbers of CFU-F in fresh AML marrow samples (nine cases studied) and the numbers of adherent nucleated cells (r = 0.91), adherent CFU-F (r = 0.87), adherent hemopoietic progenitors (r = 0.71), and colony-stimulating factor-1 (CSF-1) levels (r = 0.74) in LTMC. All the above parameters were drastically reduced in LTMC derived from AML marrows that contained low numbers of CFU-F (i.e. below the normal range of 76-130 CFU-F per 10(6) bone marrow cells) when compared with LTMC established from normal marrows. In contrast, no difference was observed in such parameters when normal LTMC and LTMC from AML patients with normal levels of marrow CFU-F were compared. Fibroblast adherent layers derived from AML patients with normal marrow CFU-F numbers produced normal levels of CSF-1 and showed a similar hemopoietic supportive capacity to that of fibroblast layers from normal bone marrow. Thus, our results suggest that (i) the presence of CFU-F in adequate levels in the bone marrow inoculum is critical for the adequate development of the hemopoietic microenvironment in LTMC, and (ii) AML marrow-derived fibroblasts seem to be functionally normal, at least in some AML patients. PMID- 8412320 TI - Expression of tie receptor tyrosine kinase in leukemia cell lines. AB - The tie receptor tyrosine kinase mRNA was originally identified as an amplified product in reverse transcription-polymerase chain reaction analysis of human K562 leukemia cell RNA. In situ hybridization analysis revealed that the corresponding mouse gene is expressed predominantly in endothelial cells. We have explored tie mRNA and protein expression in tumor cell lines. The 4.4 kb tie mRNA was expressed at high levels in five of five human megakaryoblastic leukemia cell lines studied and in two IL-3-dependent mouse myeloid leukemia cell lines, but not in 42 other leukemia cell lines representing various hematopoietic lineages. Increased expression of tie mRNA and protein was observed upon treatment of the megakaryoblastic leukemia cells with the tumor promoter 12-0-tetradecanoyl phorbol-13-acetate (TPA), known to enhance megakaryoblastic markers. Among several cell lines from solid tumors, two fibrosarcomas, one rhabdomyosarcoma and one melanoma cell line were positive for tie mRNA. These results suggest that among hematopoietic lineages tie is predominantly expressed in cells with megakaryoblastic properties and that the tie tyrosine kinase is a receptor for a regulatory factor specific for megakaryoblasts, endothelial cells, and occasional tumor cell lines derived from mesenchymal tissues. PMID- 8412321 TI - Pharmacokinetic basis for optimal hemopoietic effectiveness of homologous IL-3 administered to rhesus monkeys. AB - To design an interleukin-3 (IL-3) administration schedule for optimal hemopoietic effectiveness, serum half-life (t1/2) was determined after intravenous (i.v) and subcutaneous (s.c.) bolus injections. The initial t1/2 in serum after i.v. injection was about 10 minutes and the terminal t1/2 close to 2 hours. Subcutaneous administration resulted in plateau levels after 2 to 4 hours, while the apparent terminal t1/2 was similar to that after i.v. infusion. The bioavailability of IL-3 following subcutaneous administration was only about 40% of that following i.v. administration. Hemopoietic effects of continuous i.v. infusion of IL-3 was then compared to s.c. administration in either one, two, or three daily injections. Doses chosen ranged from 1 to 30 micrograms/kg per day. In agreement with the more limited bioavailability of IL-3 following s.c. administration, continuous i.v. infusion was much more effective in stimulating hemopoiesis than s.c. administration. Two or three daily s.c. injections did not improve the hemopoietic response compared to a single s.c. injection, which is in agreement with the apparent terminal t1/2 of 101 min. It is concluded that IL-3 is more effective by continuous i.v. infusion than by subcutaneous administration. PMID- 8412322 TI - Involvement of the complement system in the pathogenesis of pulmonary leukostasis in experimental myelocytic leukemia. AB - The role of the complement system in the pathogenesis of pulmonary leukostasis in myelocytic leukemia was studied in a rat model. Acute myelocytic leukemia was induced in six Brown-Norway rats, and complement levels were assayed during the course of the disease. Whole complement activity (CH50) and hemolytic activity of C1q, C3, and C4 decreased from day 16 after induction of the leukemia, when the rats developed pulmonary leukostasis. In addition, local complement activation was established in the lung vessels by immunofluorescence microscopy in advanced stages of pulmonary leukostasis. Finally, following systemic activation of the complement system by injection of cobra venom factor (CVF), leukemic rats (n = 6) died of pulmonary leukostasis 4.5 days earlier than did leukemic controls (n = 6). These findings suggest that, in acute myelocytic leukemia in Brown-Norway rats, pulmonary leukostasis is induced by activation of the complement system. This finding could lead to new modes of treatment for a life-threatening complication of leukemia. PMID- 8412323 TI - Establishment of a new human pre-B acute lymphoblastic leukemia cell line (KMO 90) with 1;19 translocation carrying p53 gene alterations. AB - A new human pre-B acute lymphoblastic leukemia cell line (KMO-90) was established from the bone marrow sample of a 12-year-old girl with acute lymphoblastic leukemia (ALL) carrying 1;19 chromosome translocation. KMO-90 cells expressed HLA DR, CD10, CD19, and CD22 antigens. These cells had also cytoplasmic immunoglobulin lacking surface immunoglobulin, indicating that these had a pre-B phenotype. Chromosome analysis of this cell line showed 48, XX, +8, +19, t(1;19)(q23;p13). Southern blot analysis showed the same sized rearrangements of the E2A gene in KMO-90 cells as those in the original leukemic cells. By means of reverse transcriptase-polymerase chain reaction analysis, we detected E2A/PBX1 fusion transcripts in KMO-90 cells. KMO-90 is useful when studying the role of the 1;19 translocation in the etiology of pre-B ALL. Furthermore, we studied alterations of the p53 gene in this cell line by polymerase chain reaction, single-strand conformation polymorphism analysis. KMO-90 cells were identified to have a point mutation at codon 177 (CCC-->TCC) of the p53 gene, suggesting that alterations of the p53 gene may have an important role in the establishment of this cell line. PMID- 8412324 TI - In vitro establishment of AIDS-related lymphoma cell lines: phenotypic characterization, oncogene and tumor suppressor gene lesions, and heterogeneity in Epstein-Barr virus infection. AB - Lymphoma represents a major source of morbidity and mortality among AIDS patients. AIDS-associated non-Hodgkin lymphomas (AIDS-NHL) are almost invariably B-cell derived, are classified as high or intermediate grade lymphomas, and display three main histologic types: namely, small non-cleaved cell lymphoma (SNCCL), large cell immunoblastic plasmacytoid lymphoma (LC-IBPL), and large cell lymphoma (LCL). Here we report the in vitro establishment of three new AIDS-NHL cell lines (termed HBL-1, HBL-2, and HBL-3) derived from three AIDS-SNCCL patients differing in primary tumor sites and risk factors for HIV infection. The derivation of the cell lines from the original tumor clones was established by immunophenotypic and molecular genetic analysis. These cell lines display clonal immunoglobulin gene rearrangement, express surface immunoglobulin and B-cell restricted markers, and exhibit a phenotype consistent with SNCCL. Monoclonal Epstein-Barr virus infection was found in only one of the cell lines (HBL-1). Cytogenetic analysis demonstrated the presence of a chromosomal translocation involving the c-myc proto-oncogene and an immunoglobulin locus in all three cell lines. The pattern of genetic lesions detected in HBL-1, HBL-2, and HBL-3 reflects that found in primary AIDS-SNCCL and includes activation of the c-myc oncogene as well as inactivation of the p53 tumor suppressor gene. These cell lines should prove useful in studies of the biological, immunological, and viral factors involved in AIDS-associated lymphomagenesis. PMID- 8412325 TI - Acute lymphoblastic leukaemia incidence in the UK by immunophenotype. AB - The incidence of acute lymphoblastic leukaemia (ALL) is described for both children and adults for the three major immunophenotypes: null, CD10-positive (CD+) which includes both common and pre-B types, and T-cell (including pre-T variants). The data are derived from a population-based specialist registry of leukaemias and lymphomas covering approximately one-half of England and Wales. Null ALL predominates in those under 1 year old and CD10+ ALL in the 1-7 year olds. There is a male excess at all ages for T-cell disease, which is particularly prominent in adolescents and young adults. The effect of socioeconomic levels is seen most clearly for CD10+ ALL in the childhood peak, where B-cell precursor disease occurs more frequently in areas of higher socioeconomic status. PMID- 8412326 TI - Pulsed field gel electrophoresis on single murine hemopoietic colonies. AB - We report a technique which allows for the direct molecular analysis of single whole murine hemopoietic colonies by pulsed field gel electrophoresis (PFGE). Murine bone marrow cells were plated out in semi-solid agarose and gave rise to macroscopic colonies after 11 days in culture. Single colonies were excised from the agarose using a sterile blade and embedded without further manipulation in molten low-melting-temperature agarose. The leucocyte DNA contained within the agarose plug was subjected to restriction enzyme digestion and PFGE. Sufficient high molecular weight DNA is afforded by this method to achieve a hybridization signal with a single copy probe. This method will make PFGE directly applicable to the clonal analysis of chromosomal aberrations in hemopoietic stem and progenitor cells. PMID- 8412327 TI - Molecular analysis of T-cell receptor transcripts in a human T-cell leukemia bearing a t(1;14) and an inv(7); cell surface expression of a TCR-beta chain in the absence of alpha chain. AB - The polymerase chain reaction (PCR) was used to amplify cDNA in order to characterize normal and hybrid T-cell receptor (TCR) gene rearrangements derived from a T-cell acute lymphoblastic leukemia (T-ALL) bearing a chromosome 7 inversion. The nucleotide sequence analysis of the amplified product showed the presence of an out-of-frame V beta/J gamma/C gamma transcript and an in-frame V gamma/J gamma/C beta transcript which result from an interlocus recombination between the TCR-beta and gamma loci and the transcription of the reciprocal hybrid TCR gene. The sequence analysis of the reciprocal DNA segment directly involved in the breakpoint of the inversion showed a recombination between a J gamma-sequence heptamer signal and a coding J beta gene segment. The exonuclease nibbling of each DNA extremity and the non-templated nucleotide insertion observed at both coding and reciprocal joints demonstrate that the inversion is mediated by the lymphocyte recombinase complex. During T-cell differentiation, TCR-beta genes are rearranged prior to TCR-alpha genes. In the present case of T ALL, we have shown that TCR-delta genes are rearranged at both loci, excluding productive rearrangements of TCR-alpha genes. The molecular analysis of the normal TCR genes derived from the leukemic cells revealed the presence of productively rearranged TCR-beta, gamma, and delta genes. Cell surface staining of these cells showed the presence of CD3 molecules and of a TCR-beta chain corresponding to the normal beta allele expressed in a disulfide-linked complex with a protein which is not TCR-alpha, gamma, or delta. This could represent the cell surface expression of the hybrid TCR-V gamma/C beta protein or the expression of a TCR-beta homodimer. PMID- 8412328 TI - Expression of the EVI1 gene in chronic myelogenous leukemia in blast crisis. AB - The EVI1 gene encodes a nuclear, zinc finger, DNA binding protein expressed after provirus insertion into the Fim-3 or EVI1 loci in murine leukemia myeloid cells. EVI1 is also expressed by cells from AML patients with chromosome rearrangements involving band 3q26, the putative location of the EVI1 gene, but expression of this gene is not detected in normal bone marrow. We report expression of EVI1 by cells from a patient with chronic myelogenous leukemia in blast crisis (CML/BC) whose cells showed inv(3)(q22q26). In vitro culture of these cells resulted in macrophage differentiation and loss of EVI1 expression. Results in this patient suggest EVI1 expression played a role in CML blast transformation. Patients with CML/BC and other nonrandom chromosome abnormalities involving chromosome 3q26 should be evaluated for EVI1 expression. PMID- 8412329 TI - Identification of an inversion 16 coexisting with an isochromosome 22q by in situ hybridization in a case of childhood AML M4e. AB - Rearrangements involving chromosome 16, including inv(16) (p13q22), del(16)(q22), and t(16;16)(p13;q22), are frequent findings in acute myeloblastic leukemia (AML). Each of these rearrangements can occur as the sole karyotypic change or in association with additional chromosomal abnormalities, including in decreasing order of frequency: trisomy 22, trisomy 8, and deletion of the long arm of chromosome 7. We report a pediatric case of de novo AML, M4e subtype, with a unique combination of inv(16) (p13q22) and i(22q) occurring within the same leukemic clone. The inv(16) was detected by fluorescence in situ hybridization (FISH) analysis with two cosmid probes specific for sequences flanking the inv(16) breakpoint on the long arm of chromosome 16. Use of a chromosome-22 specific painting probe unequivocally identified a small metacentric chromosome as an i(22q). This case illustrates a variation in the association of trisomy 22 with inv(16) and suggests that duplication of the long arm of chromosome 22 may contain critical gene(s) involved in the multistep process of evolution of leukemia with 16q22 abnormalities. PMID- 8412330 TI - Translocation t(1;22) mimicking t(1;19) in a child with acute lymphoblastic leukemia as revealed by chromosome painting. AB - The karyotype of a boy with acute lymphoblastic leukemia (ALL) presenting with numerical and structural chromosome aberrations as determined by Giemsa-banding was further investigated using chromosome painting (CP). A translocation t(11;18)(q23;q21) was verified by this approach, and gain of chromosome 21 material due to a structural rearrangement was detected. Moreover, an unbalanced translocation of the long arm of chromosome 1, resembling the well known translocation t(1;19), was demonstrated to involve chromosome 22 instead of chromosome 19. Immunophenotyping of the leukemic blasts led to the diagnosis common ALL (CD19+, CD10+, clg-). Our case indicates that in ALL a translocation t(1;19) may be mimicked by other chromosomal rearrangements, and that CP may efficiently complement conventional cytogenetics in the exact characterization of the involved chromosomes. PMID- 8412331 TI - Transient myeloproliferative disorder in a Down's neonate with rearranged T-cell receptor beta gene and evidence of in vivo maturation demonstrated by dual-colour flow cytometric DNA ploidy analysis. AB - Neonates with Down's syndrome may develop a transient myeloproliferative disorder (TMPD) which on presentation is indistinguishable from acute leukemia, with the difference manifest only on follow-up. The clinical course is one of spontaneous remission in TMPD and relentless progression in leukemia. We describe a Down's neonate presenting with hyperleucocytosis and circulating blasts which were positive for CD34, myeloid (CD33), megakaryocytic (CD41, CD42b, CD61), and T lineage (CD3, CD7), but not B-lineage, associated antigens. Clonal rearrangement of the T-cell receptor beta (TCR beta) gene was found, with the immunoglobulin heavy chain gene in germline configuration, showing the disease to be a clonal proliferation of a multipotential stem cell involving the myeloid and T lineages. Dual-colour flow cytometric DNA ploidy analysis of CD41 positive blasts showed initially a predominant 2N population, but polyploidization to 6N and 8N cells was found on follow-up, concomitant with a progressive decrease in circulating blasts, suggesting maturation of the abnormal clone and a provisional diagnosis of TMPD. This was shown by the eventual resumption of normal haemopoiesis with the disappearance of blasts and the clonally rearranged TCR beta gene. PMID- 8412332 TI - Hypercalcemia in the blastic phase of chronic myeloid leukemia associated with elevated parathyroid hormone-related protein. AB - We describe a case of hypercalcemia without lytic bone lesions complicating myeloid blast crisis of chronic myeloid leukemia (CML). Serum levels of parathyroid hormone-related protein (PTHrP) were elevated during the initial hypercalcemic period and became undetectable during chemotherapy-induced chronic phase, only to become elevated again during subsequent recurrent blastic periods repeatedly associated with hypercalcemia. Previously reported cases of hypercalcemia complicating CML are reviewed. It is suggested that PTHrP was responsible for the hypercalcemia in this case and may be an important mediator of hypercalcemia in CML. PMID- 8412333 TI - The pediatric rhabdomyosarcoma translocation (2;13)(q35;q14) in B-prolymphocytic leukemia. PMID- 8412334 TI - Therapy-related acute promyelocytic leukemia. PMID- 8412335 TI - Urinary excretion of protein and non-protein hydroxyproline in patients with sarcoidosis and interstitial pulmonary fibrosis. AB - The determination of total hydroxyproline (Hyp) with separation into protein and non-protein fractions was carried out in 24-hour urine samples of healthy persons, patients with sarcoidosis and patients with interstitial pulmonary fibrosis (i.p.f.). In the group of sarcoidosis Hyp excretion was evaluated taking into account progression of pulmonary changes and disease activity. It was found that total and non-protein Hyp excretion correlated positively in healthy persons as well as in those with sarcoidosis, and interstitial pulmonary fibrosis. An increased excretion of total and non-protein Hyp occurred in 24% of the patients with sarcoidosis while excretion of protein Hyp remained within the normal range. In 30% of the patients with interstitial pulmonary fibrosis increased excretion of total and non-protein Hyp was demonstrated. The excretion of protein Hyp in these patients did not deviate from normal values. The results of our study indicate that determination of total and nonprotein Hyp in urine may be useful in the evaluation of fibrotic changes in the pulmonary diseases mentioned above. However, these parameters are not usefull in differentiation between sarcoidosis and interstitial pulmonary fibrosis. PMID- 8412336 TI - An attempt at acoustic estimation of the results of phonosurgery. AB - The acoustic method of voice examination is presented. The method was verified in the group of 62 patients. The examination was performed before and after larynx microsurgery. All results were compared with traditional examination of voice. The method revealed a good sensitiveness as well as reliability and seems to be useful for monitoring of patients after phonosurgery. PMID- 8412337 TI - Changes of certain biochemical placental components during Salbupart tocolysis. AB - The studies material comprised 10 placentae from pregnancies at risk for premature labour with delayed intrauterine growth of the fetus treated with Salbupart from the 32nd week of pregnancy. In placental homogenates the biochemical components were determined for fragmentary assessment of carbohydrate and lipid metabolism and production on high-energy compounds. Salbupart was found to increase the oxidation processes in the placenta. This increases the production of biologically useful chemical energy which is expressed as increased energy charge (so called Atkinson's index or adenylate energy charge). Increased energy yield of the placenta results in improved materno-fetal transport and contributes to greater weight gain of the hypotrophic fetus. PMID- 8412338 TI - Plasma insulin and blood pressure response to oral glucose tolerance test in young borderline hypertensives. AB - In order to evaluate whether borderline hypertension might be associated with hyperinsulinaemia, twenty non-obese borderline hypertensives and twenty matched normotensives underwent a standard oral glucose tolerance test and 24-h ambulatory blood pressure monitoring. Blood pressure, plasma glucose and insulin were measured at fasting and 15, 30, 60, 120 and 180 minutes after glucose load. Fasting plasma insulin was significantly higher in borderline hypertensives in comparison to normotensives (16.6 +/- 6.9 vs 12.4 +/- 4.2 mU/l; P < 0.05). Plasma insulin response estimated by the positive incremental area under the curve did not differ significantly between two groups but borderline hypertensives showed a larger interindividual difference. Decrease of systolic blood pressure after glucose load was significantly greater in borderline hypertensive subjects. Furthermore, blood pressure and plasma insulin relationship was different in borderline hypertensives compared to normotensives. PMID- 8412339 TI - Morphology of canine pancreas in electron microscopy following truncal and highly selective vagotomy. AB - The purpose of the study was evaluation of morphological changes in canine pancreas on days 30, 90 and 180 after truncal vagotomy (VT) and highly selective vagotomy (HSV). On the 30th day after VT in some cells zymogen granules and lysosomes were disintegrated, vacuolization was moderately expressed, and fibrosis was present in the exocrine part. On the 30th day after HSV the structure of the exocrine part showed no important changes, but in the endocrine part in about one-third of cells in the islets deformed nuclei and trinucleated cells were found, along with changes of mitochondrial structure. beta cells contained very numerous insulin-containing granules. On the 90th day some changes observed on the 30th day after VT or HSV were no longer seen. On the 180th day after HSV the EM changes were negligible, less intense than after VT. The electron microscopic changes after VT and HSV were most pronounced early after the operation, particularly after VT. In later stages of the experiment a part of the changes regressed, but then more structural changes were seen after VT than after HSV. PMID- 8412340 TI - Inhibition of lipid peroxidation in rat brain by nifedipine and clorazepate after electrically induced seizures. AB - The effect of nifedipine and clorazepate on the concentration of lipid peroxides (LP) in rat brain, and the characteristics of electrically induced seizures were assessed. A significant increase in the concentration of brain LP after electroshock was found. Both nifedipine (1.00 mg/kg per os) and clorazepate (20 mg/kg intraperitoneally) decreased the levels of LP in the rat brain after electroshock. Nifedipine combined with clorazepate brought an inhibition of LP formation and an additive anticonvulsant activity. PMID- 8412341 TI - Free radicals and antioxidant systems. AB - Free radicals are undesirable peroxides that have long been implicated in several deleterious effects in our body including aging process. They also play an important role in many physiological processes. For they are very reactive in nature, there are some anti-oxidant systems to scavenge or to quench them. PMID- 8412342 TI - Establishing requirements for the use of terms for reporting adverse drug reactions (ADR). Consultant, Council for International Organizations of Medical Sciences (CIOMS), World Health Organization. PMID- 8412343 TI - Spontaneous perforation of rectum and rectosigmoid junction. Case report. AB - Two patients with spontaneous rectoisigmoid or rectal rupture are reported. These patients had peritonitis and they underwent sigmoid loop colostomies. A common aetiological factor was chronic constipation. It is important to recognize the condition at an early stage because of its significant mortality and possible excellent results from surgery. PMID- 8412344 TI - Leucocyte ascorbic acid concentration and plasma ascorbic acid levels in children with various infections. AB - In this study leucocyte ascorbic acid concentrations and plasma ascorbic acid levels were measured simultaneously and correlation between these two variables was sought in various bacterial and viral infections. A total of 258 patients were studied. The control group consisted of 21 healthy children in the same age group. Except for scarlet fever no significant differences were observed between the patients' leucocyte ascorbic acid levels during and after the infections and also no differences were observed in leucocyte ascorbic acid levels between control's and patient's. However plasma ascorbic acid values were significantly decreased during infection in all patients. During infection ascorbic acid is utilized rapidly, and our findings seem to indicate that there is transfer of ascorbic acid from plasma to leucocytes in order to keep the leucocyte ascorbic acid levels within normal limits. PMID- 8412345 TI - Rat liver pathomorphology during prolonged sodium valproate administration. AB - Prolonged administration (1, 3, 6, 9 and 12 months) to rats of an antiepileptic drug--sodium valproate (Vupral--"Polfa") in the dose of 200 mg/kg/day produced the first hepatic morphological lesions after 3 months of the experiment. These structural abnormalities progressively increased to achieve their peak within 12 months. The most prominent microscopic changes consisted of extensive microvesicular fatty change and vacuolar degeneration of periportal hepatocytes. Intralobular focal necrosis, infiltrations of lymphocytes, plasma cells, and phagocytes in the portal tracts, and centrilobular congestion were also present. The connective tissue did not proliferate either in periportal lobular zone or around the central veins. The described lesions seem thus to be reversible. PMID- 8412346 TI - Child maltreatment: international perspectives. AB - The purpose of this literature review was to explore definitions, incidence, and management of child maltreatment across cultures. Articles written in the English language published from 1962 to 1991 were reviewed to answer the following questions: (1) What role does cross-cultural variability play in defining child maltreatment? (2) What is the incidence of maltreatment in developed and developing countries across continents? (3) What measures have been instituted by countries to prevent and manage child maltreatment? Cross-cultural information was found to be limited. Child rearing attitudes had an impact on the identification, prevention, and management of maltreatment across nations. PMID- 8412347 TI - High-risk infant case management and assistive technology: funding and family enabling perspectives. AB - Nursing personnel who are high-risk infant case managers need to be aware of potential funding resources in order to provide needed technology and services to families of high-risk infants and young children. The authors review federal funding streams for the provision of assistive technology and services for this population. Emphasis is placed on encouraging parents to function as independently as possible and assume active rather than passive roles in the case management process. Practical strategies for assisting families to secure needed funding are presented, including the development of a funding portfolio, obtaining technology evaluations, gathering documentation, and becoming involved in appeal processes. PMID- 8412348 TI - Self-described learning needs of pregnant teen participants in an innovative university/community partnership. AB - The author of this descriptive study investigated self-described learning needs and preferences for teaching methods of pregnant 7th-12th-grade students in an alternative school setting who participated in health classes taught by nursing students. Subjects (N = 121) completed the Learning Needs Assessment for Pregnant Adolescents questionnaire. Results indicated that teens were most interested in learning about infant illnesses, along with complications during pregnancy and labor and delivery. The Evaluation of Health Class tool indicated that teaching strategies preferred by subjects (n = 83), were hospital visits, parties, videotapes, films, records, and games. These results suggest that teaching topics and strategies are important considerations when providing health information for pregnant teens. PMID- 8412349 TI - Sir Andrew Huxley--1963 Nobel Prize Winner. PMID- 8412350 TI - Ocular adnexal lymphoproliferative lesions. AB - Ocular adnexal lymphoproliferative lesions consist of a spectrum of disease entities, including reactive lymphoid hyperplasia, atypical lymphoid hyperplasia, and lymphoma. No clinical or radiologic criteria facilitate a distinction among these lymphoproliferative lesions. The two hyperplastic processes may evolve to localized or systemic lymphoma. A similar pattern is evident in other mucosa associated lymphoid tumors elsewhere in the body. Most ocular adnexal lymphomas are small lymphocytic non-Hodgkin's tumors with an indolent course; frequently, they remain localized to the ocular adnexa. In comparison, intermediate- and high grade lymphomas are less common in the ocular adnexa but more aggressive. An approach to the diagnosis and treatment of these complex entities is suggested. Despite new pathologic classification schemes, immunophenotypic labeling, and molecular genetic analysis, the prognosis for patients with small-cell lymphoma in the ocular adnexa is difficult to predict. PMID- 8412351 TI - Intravitreal large-cell lymphoma. AB - Large-cell (non-Hodgkin's) lymphoma may occur in the eye as a cellular infiltrate in the vitreous, uveal tract (choroid), retina, or optic nerve. Lymphomatous involvement may be limited to the eye but also is frequently associated with lesions of the central nervous system. Ocular involvement may precede involvement of the central nervous system by months or, in some cases, years. Ocular large cell lymphoma is bilateral in approximately 80% of cases but often is asymmetric. The mean age of patients with ocular large-cell lymphoma is 60 years, and women are affected almost twice as often as men. Intravitreal large-cell lymphoma may manifest as an infiltrate of large glassy-gray cells or clusters of cells, and it may mimic uveitis or other inflammatory and infectious conditions of the eye. The diagnosis is based on cytologic and immunocytochemical studies of a vitreous biopsy specimen obtained by aspiration or by vitrectomy through the pars plana. Advances in irradiation of the eyes and the central nervous system, supplemented with corticosteroids and intrathecally and intravenously administered chemotherapeutic agents, have resulted in improvement of the dismal prognosis for patients with large-cell lymphoma. PMID- 8412352 TI - Prolonged QT syndrome: perioperative management. AB - Intraoperative cardiac arrhythmias related to prolonged QT syndrome are uncommon. We describe a 26-year-old woman in whom ventricular fibrillation occurred during the final minutes of a partial glossectomy and right supraomohyoid selective neck dissection and discuss the role that this specific operation may have had in the development of the intraoperative event. In addition, we review the perioperative management of patients with prolonged QT syndrome. PMID- 8412353 TI - Ultrafast computed tomographic scanning to assess patency of coronary artery stents in bypass grafts. AB - Anginal chest pain after implantation of coronary stents in bypass grafts is a concern because it suggests the possibility of occlusion. Coronary angiography is the definitive method for determining patency of a stent; however, this procedure is relatively contraindicated in a patient receiving warfarin sodium, who has a therapeutic international normalized ratio. An alternative method for determining patency of a stent is by ultrafast computed tomography. This new, minimally invasive technique shows promise for determining blood flow within the large vessels of the thorax. Herein we describe a case in which a metallic stent placed in a vein graft was noninvasively established to be patent, despite chest pain in the patient. This method may be clinically applicable for determining patency of stents in vein grafts in the setting of patients with chest pain who have undergone complete anticoagulation. PMID- 8412354 TI - Achalasia, aspiration, and atypical mycobacteria. PMID- 8412355 TI - Leukocyte differential. PMID- 8412356 TI - Psychology in sports medicine. PMID- 8412357 TI - Can symptoms associated with dysphagia be of discriminate value? PMID- 8412358 TI - Dr. Donald Church Balfour--sixth Mayo partner, surgeon, and educator. PMID- 8412359 TI - Competitive athletes: preinjury and postinjury mood state and self-esteem. AB - In a prospective study, we determined whether preinjury and postinjury differences existed in the mood state and self-esteem of competitive athletes. The influence of severity of injury, gender, level of participation in sports, and type of sport on these dependent variables was also measured. Among 238 male and 38 female athletes from hockey, basketball, baseball, and volleyball teams, 36 sustained 43 injuries. Significant postinjury increases were noted for depression (P < 0.0001) and anger (P = 0.0012), whereas vigor (P < 0.0001) was significantly less after injury. When the 36 injuries were classified, 27 were minor or moderate (nonparticipation in sports for only one or two weekly assessments), and 9 were severe (nonparticipation for three or more weekly assessments). When a stepwise multiple regression equation was used to predict the scores for postinjury depression, the only significant predictor was the severity of injury (F = 8.48 [1, 34]; R2 = 0.30; P = 0.0063). Of the following physical and psychosocial variables--level of participation, type of sport, age, previous injury, preinjury stress, gender, mood state scales, and self-esteem- only level of participation (P < 0.0001) and type of sport (P = 0.0004) were predictors of injury. The significant preinjury and postinjury differences in mood state suggest that postinjury mood disturbances reported in previous studies are likely attributable to the occurrence of injury, are related to the severity of injury, and do not merely reflect a disturbed preinjury mood. PMID- 8412360 TI - Discriminate value of esophageal symptoms: a study of the initial clinical findings in 499 patients with dysphagia of various causes. AB - The aim of this study was to determine whether the characteristics of esophageal symptoms may be of diagnostic utility in distinguishing dysphagia of various causes. Included in the study were a total of 499 patients with three types of dysphagia: 234 with dysphagia related to an obstructive lesion in the esophagus, 162 with dysphagia related to disturbed esophageal motility, and 103 with dysphagia who had no demonstrable structural or motor abnormalities in the esophagus. In the first part of the study, the initial esophageal symptoms of 402 patients with dysphagia were retrospectively reviewed and analyzed with use of a stepwise logistic regression. This analysis led to development of models that could distinguish among the various diagnostic groups of dysphagia. In the second segment of the study, these models were validated in a prospective assessment that involved 97 patients with dysphagia. In the third part of the study, the models were further refined by using data from all 499 patients from the first two parts. The results presented herein suggest that a subset of selected esophageal symptoms can distinguish among diagnostically identified groups of dysphagia. The discriminate models presented rely on a few easily determined clinical variables and hence are practical and potentially useful in the evaluation of undifferentiated dysphagia. PMID- 8412361 TI - Simultanagnosia as the initial sign of degenerative dementia. AB - In a study of 10 patients with degenerative brain disease that manifested as simultanagnosia, our aims were (1) to elucidate their clinical, neuropsychologic, and radiologic findings to determine whether these patients might represent a group distinguishable from those with typical Alzheimer's disease and (2) to help clinicians recognize this entity. All patients were initially examined by ophthalmologists because of visual difficulties, and the simultanagnosia remained undiagnosed until nonophthalmologic complaints developed. Optic ataxia developed in six patients, and all patients had mildly impaired eye movements. All 10 patients could identify colors appropriately. Nine patients had language deficits (anomia, decreased auditory comprehension, alexia, and agraphia) but were fluent and had relative preservation of sentence repetition, and four performed in the normal range on a test of associative fluency. Two patients scored in the normal range on memory tests, all had preserved insight, and nine had no family history of degenerative dementia. The mean age at onset of the disorder was 60 years (range, 50 to 69). Neuroimaging disclosed prominent bilateral occipitoparietal atrophy in nine patients and generalized atrophy in one. With this unusual but consistent clinical, neuropsychologic, and anatomic profile, these patients are clinically distinguishable from those with typical Alzheimer's disease, but until a specific cause has been found, we cannot be certain that they constitute a specific biologic entity. Clinicians should consider this diagnosis in relatively young patients who have slowly progressive nonocular visual complaints. PMID- 8412362 TI - Stereotactic procedures for lesions of the pineal region. AB - During the 7-year period between June 1985 and May 1992, 34 patients with pineal lesions underwent 66 stereotactic procedures (37 biopsies, 19 third ventriculostomies, 6 cyst aspirations, 3 instillations of 32P into cysts, and 1 insertion of an Ommaya reservoir into a cyst) at the Mayo Clinic. Nine patients subsequently also underwent 10 open resections of lesions of the pineal region. In the 34 study patients, the pathologic entities were 9 gliomas (5 astrocytomas, 2 ependymomas, and 2 oligodendrogliomas), 9 germ cell tumors (7 germinomas, 1 entodermal sinus tumor, and 1 malignant teratoma), 8 pineal parenchymal tumors (3 pinealomas, 3 pinealoblastomas, 1 mixed pinealoma-pinealoblastoma, and 1 intermediate differentiation pineal tumor), 4 other malignant tumors (2 undifferentiated carcinomas, 1 malignant melanoma, and 1 non-Hodgkin's lymphoma), 2 meningiomas, and 2 nonneoplastic lesions (1 glial cyst and 1 inflammatory lesion). No mortality or permanent morbidity was associated with the 66 stereotactic procedures; 2 patients had temporary complications--1 neurologic (transient diplopia) and 1 nonneurologic (pulmonary embolism). Diagnostic tissue was obtained in 33 of the 34 patients. An algorithm for the diagnosis and management of patients with lesions of the pineal region is presented. We conclude that stereotactic biopsy of pineal lesions can be performed safely, has a high diagnostic yield, and facilitates rational planning of treatment. PMID- 8412363 TI - Aging and the human pituitary gland. AB - Pituitary glands obtained at autopsy from 41 men (90 to 97 years old) and 45 women (90 to 98 years old) were studied histologically and immunohistochemically to determine the age-related alterations. Pituitaries from patients 30 to 39 years old (48 cases) and 60 to 89 years old (187 cases) were also studied and served as controls. Interstitial, perivascular fibrosis was seen in 88% of the aged adenohypophyses and, although significantly more intense than that in those of the fourth decade of life (P < 0.001), was similar to that noted in those of the seventh to ninth decades. Pituitaries of men were significantly more fibrotic than were those of women (P < 0.05). In proportion to the extent of fibrosis, the number of somatotrophs decreased in the lateral wings, whereas other cell types did not change quantitatively. Small deposits of amyloid and of iron were detected in seven and three cases, respectively. Squamous metaplasia in cells of the pars tuberalis was noted in 29%. The incidence of "basophil invasion" (the presence of corticotrophs in the posterior lobe) was 30%, a figure similar to that in younger controls. Granular cell tumorlets were detected in four aged neurohypophyses (5%), a frequency similar to that in control glands. Pituitary adenomas (two null cell, two lactotroph, and one corticotroph adenomas) were found of three men and two women in the study group. This 9% incidence of adenomas did not differ from that observed in the fourth decade (8%) or seventh decade (10%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412364 TI - Chronic pouchitis after ileal pouch-anal anastomosis: responses to butyrate and glutamine suppositories in a pilot study. AB - Nonspecific, idiopathic inflammation of ileal pouch mucosa ("pouchitis") after ileal pouch-anal anastomosis is a common complication of this surgical approach. The epithelium of the pouch is ileal, but variable degrees of colonic metaplasia are natural sequelae of construction of such a pouch. One hypothesis is that pouchitis is caused by a deficiency of epithelial nutrition. Thus, a lack of butyric acid (the principal metabolic fuel of colonocytes) or of glutamine (the main fuel of enterocytes) may develop. In this study, our aims were to determine the concentration of total short-chain fatty acids in random stool samples obtained from patients with an ileal pouch-anal anastomosis with and without pouchitis and to test the therapeutic effects of butyrate and glutamine suppositories on pouchitis. During the study, all conventional therapy for pouchitis was discontinued. For 21 days, 9 patients participated in a butyrate trial, and 10 patients were treated with glutamine. Total concentrations of fecal short-chain fatty acids were significantly less in patients with pouchitis than in those without pouchitis. During treatment, 6 of the 10 patients who received glutamine had no recurrence of symptoms, but only 3 of the 9 patients who received butyrate responded similarly. Hence, further studies of glutamine in the treatment of pouchitis seem warranted. PMID- 8412365 TI - Transport of the critically ill child. AB - During the past several decades, the transport of critically ill patients to and between hospitals has gradually improved. The major indications that necessitate emergency transport for adult patients are trauma and acute cardiac disease, and the establishment of transport teams trained in the care of these conditions has improved the outcome of adult patients. In critically ill children, the indications for emergency transport differ from those in adults; pediatric patients are most likely to be transported for respiratory and neurologic emergencies. The outcome for critically ill children is optimal when a dedicated pediatric transport team functions in the setting of a regional critical-care outreach program. Such a program emphasizes stabilization of the child's condition at the local hospital, followed by transport to a pediatric intensive care facility by a specially trained pediatric transport team. In this review, we present an overview of the principles and operating procedures of such pediatric transport teams. PMID- 8412366 TI - The central autonomic network: functional organization, dysfunction, and perspective. AB - The central autonomic network (CAN) is an integral component of an internal regulation system through which the brain controls visceromotor, neuroendocrine, pain, and behavioral responses essential for survival. It includes the insular cortex, amygdala, hypothalamus, periaqueductal gray matter, parabrachial complex, nucleus of the tractus solitarius, and ventrolateral medulla. Inputs to the CAN are multiple, including viscerosensory inputs relayed on the nucleus of the tractus solitarius and humoral inputs relayed through the circumventricular organs. The CAN controls preganglionic sympathetic and parasympathetic, neuroendocrine, respiratory, and sphincter motoneurons. The CAN is characterized by reciprocal interconnections, parallel organization, state-dependent activity, and neurochemical complexity. The insular cortex and amygdala mediate high-order autonomic control, and their involvement in seizures or stroke may produce severe cardiac arrhythmias and other autonomic manifestations. The paraventricular and other hypothalamic nuclei contain mixed neuronal populations that control specific subsets of preganglionic sympathetic and parasympathetic neurons. Hypothalamic autonomic disorders commonly produce hypothermia or hyperthermia. Hyperthermia and autonomic hyperactivity occur in patients with head trauma, hydrocephalus, neuroleptic malignant syndrome, and fatal familial insomnia. In the medulla, the nucleus of the tractus solitarius and ventrolateral medulla contain a network of respiratory, cardiovagal, and vasomotor neurons. Medullary autonomic disorders may cause orthostatic hypotension, paroxysmal hypertension, and sleep apnea. Neurologic catastrophes, such as subarachnoid hemorrhage, may produce cardiac arrhythmias, myocardial injury, hypertension, and pulmonary edema. Multiple system atrophy affects preganglionic autonomic, respiratory, and neuroendocrine outputs. The CAN may be critically involved in panic disorders, essential hypertension, obesity, and other medical conditions. PMID- 8412367 TI - Body composition of physically inactive and active 25-month-old female rats. AB - The purpose of this study was to evaluate the effects of physical activity on the body composition of ageing female rats. Female pathogen-free Long-Evans rats were housed either in individual 7 x 14 x 8 inch cages or in cages with attached running wheels to which they had free access. The runners ate significantly more than the sedentary rats. Food intake from month 10 through month 24 of age averaged 14.6 +/- 0.7 g for the sedentary group, and 18.3 +/- 2.2 g for the active group. The body fat content of the sedentary rats was approximately 50% higher, while their lean body mass and protein content were significantly lower than that of the runners at age 25 months. Total body weight was similar in the active and sedentary groups. Percent body fat and protein of the 25-month-old physically active rats were not significantly different from that of 9-month-old rats, while the sedentary 25-month-old rats had a significantly higher body fat content and a lower body protein content than the 9-month-old animals. These results suggest the possibility that the changes in body composition that occur during middle age in sedentary female rats are largely due to physical inactivity, and that the lean tissue wasting that occurs as the result of the aging process is a late event that occurs closer to the end of life. PMID- 8412368 TI - A step in the process of prostacyclin production whose decline leads to the age related decrease in production by human umbilical vein endothelial cells in culture. AB - Human umbilical vein endothelial (HUVE) cells were examined at various culture ages for four steps of prostacyclin (PGI2) production ((1) incorporation of exogenous arachidonic acid into the cell membrane, (2) loss of arachidonic acid from the cell membrane to the cytoplasm, (3) conversion of arachidonic acid to PGI2 in the cytoplasm, (4) release of PGI2 to the outside of the cell) to determine the step whose decline leads to the age-related decrease in PGI2 production. Only conversion to PGI2 (step 3) showed the age-related decrease. Judging from these results, the authors conclude that the age-related decrease in PGI2 production is mainly due to the decrease in the ability of HUVE cells to convert arachidonic acid to PGI2 in the cytoplasm. PMID- 8412369 TI - Phorbol ester PMA stimulates protein synthesis and increases the levels of active elongation factors EF-1 alpha and EF-2 in ageing human fibroblasts. AB - Phorbol esters modulate gene expression, reorganize the cytoskeleton and stimulate bulk protein synthesis and the steps of initiation and elongation. We have observed that a phorbol ester PMA stimulates protein synthesis and increases the amounts of active elongation factors, EF-1 alpha and EF-2 in cultured human fibroblasts MRC-5 undergoing ageing. Although bulk protein synthesis slows down during ageing, the cellular response to the stimulatory effects of PMA is higher in senescent cells. Similarly, despite the age-related decline in the amounts of active EF-1 alpha and EF-2, senescent cells exhibit a higher response to PMA. The results indicate an age-dependent increase of cellular responsiveness to PMA and provide evidence about both the integrity of the translational apparatus and the effectiveness of the signal transduction pathways during cellular ageing. In comparison, the effects of PMA on SV40-transformed MRC-5V2 cells were minimal. PMID- 8412370 TI - The effects of aging on the function of alveolar macrophages in mice. AB - In order to determine whether the function of alveolar macrophages (AM) is modulated by aging, we measured the TNF-alpha production, phagocytic function, and surface antigen expression of AM from young and old mice. When AM were primed by IFN-gamma (500 units/ml) and triggered by LPS (100 micrograms/ml), TNF-alpha production by AM was significantly smaller in old mice as compared with young mice (young mice: 161.7 +/- 28.2 units/ml; old mice: 89.3 +/- 13.6 units/ml, P < 0.05). The percentage of AM which phagocytosed latex particles (more than one particle) in old mice was significantly lower than in young mice (young: 78.1 +/- 2.5%; old: 62.8 +/- 3.4%, P < 0.05). Ia antigen expression of the AM was significantly higher and asialo-GM1 antigen expression was significantly lower in old mice than in young mice (Ia: young, 0.030 +/- 0.005; old, 0.092 +/- 0.024, P < 0.05; asialo-GM1: young, 0.-9 +/- 0.01; old, 0.75 +/- 0.07, P < 0.01). These results suggest that alveolar macrophage function is at least decreased in part with aging in mice. PMID- 8412371 TI - Age, as well as cell turnover kinetics, regulates brain/gut repair. AB - The prototypical brain/gut peptide cholecystokinin (CCK) has been used to assess brain and gut repair kinetics following cytotoxic injury in the rat. Studies addressed the effect of repetitive injury as well as aging. Injury was induced by one of the two alkylating agents, one active in the brain, the other systematically. Consistently the responses differ between brain and intestine. Total RNA falls (as predicted) in the intestine (control 1.5 +/- 1.4 versus cytotoxic 0.21 +/- 0.06 tRNA mg/organ, P < or = 0.0001), but rises (unexpectedly) in the brain (control 0.79 +/- 0.04 versus cytotoxic 1.02 +/- 0.03 tRNA mg/organ, P < or = 0.001). CCK mRNA concentration falls in the brain (predicted) (control 27 +/- 1 versus cytotoxic 11 +/- 1 pg CCK mRNA/micrograms tRNA, P < or = 0.001), but rises in the intestine (unexpectedly) (control 0.18 +/- 0.02 versus cytotoxic 0.3 +/- 0.04 pg CCK mRNA/micrograms tRNA, P < or = 0.001). CCK peptides do not change in the brain (control 39 +/- 4 versus cytotoxic 34 +/- 4 nmol/g, P < or = NS), but rise (unexpectedly) in the intestine (control 43 +/- 4 versus cytotoxic 250 +/- 27 nmol/g, P < or = 0.001). We ascribe these observations to differing brain/gut cell turnover kinetics. These data indicate that a rebound phenomenon occurs during gut recovery from cytotoxic injury. We additionally show a differential age-related response to cytotoxic injury. Younger animals tolerated the injury better than old ones (mortality: young 27% (3/11) versus old 66% (8/12), P < or = 0.001). Additionally, intestinal recovery is more rapid in younger animals. These data suggest that with increasing age, chemotherapeutic dosages may need to be modulated. It is additionally possible that clinically applicable algorithms may be developed using our animal model. PMID- 8412372 TI - Ocular microtremor measurement system: design and performance. AB - The frequency of ocular microtremor (OMT) is related to the functional status of the brain stem, and thus OMT may be useful in the diagnosis and management of brain stem disorders. The paper discusses the design of an OMT measurement system and reports quantitative specifications for three portable systems. All systems use a piezo-electric element as the transducer, which measures the displacement of the sclera during eye rotations. The systems differ in the manner in which the signal is recorded. All systems can detect eye movements corresponding to displacements of the sclera ranging from 12 to over 3000 nm. The frequency responses of all systems are flat (< 2 dB deviation from peak response) between 20 and 150 Hz. The phase response shows deviations (< pi) at the extremes of this range, but qualitative comparison of input and measured signals demonstrates that phase distortion is not excessive. Thus all systems are acceptable for clinical studies involving OMT. PMID- 8412373 TI - Treatment of chronic wounds by means of electric and electromagnetic fields. Part 2. Value of FES parameters for pressure sore treatment. AB - Subjects with spinal cord injury are often distressed by pressure sores, which usually appear after prolonged pressure (wheelchair, bed) across the soft tissue which has already lost sensibility and has diminished microcirculation. The healing ability and its dynamics depend on the state of the subject's overall health. Consequently, evaluation of a particular treatment requires careful consideration of as many as possible of the parameters relevant to healing and an adequate criterion for assessing the state of the pressure sore. Bearing in mind these two circumstances, the results of a multicentre clinical study are analysed. The aim of the study was to test two hypotheses: first that healing is faster when sores are also treated by electric currents (ECs) (in addition to conventional treatment); and secondly that there exist differences in the efficiency of the treatment if direct or low-frequency pulsed currents (FES parameters) are applied. The data analysed show that pressure sores are likely to heal twice as fast when treated with low-frequency pulsed currents. EC seems to improve the healing rate in cases where the natural healing mechanisms of the body are not sufficient (chronic wounds, older subjects). PMID- 8412374 TI - Computer analysis of heart rate variation and breathing movements in fetal lambs. AB - A quantitative method for studying the frequency-specific relationships between heart rate (HR) and fetal breathing movements (FBM) was developed. The reactivity of periodic HR variation in relation to FBM was investigated by means of power spectral analysis. Seven fetal lambs were studied during the third trimester of gestation using a chronic animal model. HR variability increased at the rate of FBM, as shown by an increase of spectral density at > 0.35 Hz in the HR autospectrum and in the cross-spectrum of HR and respirogram, as well as by an increase in the short-term variability index CVS. FBM were associated with the increased HR variation in all but the lowest frequency bands (0.07-1.0 Hz). Although respiratory sinus arrhythmia was found, only 10 per cent of the total HR variability and 25 per cent of the joint-density of HR and respirogram appeared at > 0.35 Hz during FBM. The greatest variation in both the HR and respirogram spectra appeared at < 0.07 Hz. Although the low-frequency variability of HR and respirogram was simultaneous, it was on the whole not synchronised. The existence of multiple control systems that simultaneously link the cardiac and respiratory control mechanisms to each other in the fetal lamb is postulated. PMID- 8412375 TI - Comparison of spectral techniques for computer-assisted classification of spectra of heart sounds in patients with porcine bioprosthetic valves. AB - The diagnostic performance of two spectral techniques (the fast Fourier transform, FFT, and autoregressive modelling, ARM) combined with four windowing functions (rectangular, Hanning, Hamming, and sine-cosine) and two classifiers (Bayes and nearest neighbour) to detect valvular degeneration was evaluated in a group of 95 patients. Forty-seven patients had a porcine bioprosthetic valve inserted in the aortic position and 48 patients had a porcine bioprosthetic valve inserted in the mitral position. Among the aortic valves, 24 were normal and 23 were degenerated whereas among the mitral valves, 19 were normal and 29 were degenerated. The aortic and mitral valves were analysed separately. For each type of valve, 21 features were extracted from the spectra of the valve closure sounds to train and test the performance of four pattern recognition systems by using the leave-one-out method. The discriminant properties of all feature combinations between two and five among the 21 features selected were evaluated. Results show that the FFT combined to the nearest neighbour classifier provided the best performances: 87 per cent of correct classifications (CCs) for aortic valves when using the Hanning or the Hamming window and 94 per cent of CCs for mitral valves when using the rectangular window. The best performances obtained with the ARM were 81 per cent of CCs for the aortic valves (nearest neighbour classifier and the Hanning or the Hamming window) and 92 per cent of CCs for the mitral valves (nearest neighbour classifier and the Hamming window or the Bayes classifier and the Hanning or the Hamming window). PMID- 8412376 TI - Cardiac Doppler blood-flow signal analysis. Part 1. Evaluation of the normality and stationarity of the temporal signal. AB - The normality (Gaussian property) and stationarity of the cardiac Doppler blood flow signal were evaluated on short-time segments distributed over the cardiac cycle. The basic approaches used to perform statistical tests on the nonstationary and quasiperiodic cardiac Doppler signal are presented. The results obtained from the data of ten patients having a normal aortic valve and ten patients having a stenotic valve indicate that a complex Gaussian random process is an acceptable approximation for the clinical cardiac Doppler signal. For segments of 10 ms or less, 82 per cent of them were accepted to be stationary with a significance level of 0.05, whereas for durations greater than 40 ms, the percentage of stationary segments was less than 75 per cent. It was concluded that the 10 ms window generally used in practice is a good choice for Doppler spectrogram estimation, but a shorter time interval would be preferable. PMID- 8412377 TI - Cardiac Doppler blood-flow signal analysis. Part 2. Time/frequency representation based on autoregressive modelling. AB - Doppler spectrograms obtained by using autoregressive (AR) modelling based on the Yule-Walker equations were investigated. A complex AR model using the in-phase and the quadrature components of the Doppler signal was used to provide blood flow directions. The effect of model orders on the spectrogram estimation was studied using cardiac Doppler blood flow signals taken from 20 patients. The 'final prediction error' (FPE) and the 'Akaike's information criterion' (AIC) provided almost identical results in model-order selection. An index, the spectral envelope area (SEA), was used to evaluate the effect of window duration and sampling frequency on AR Doppler spectrogram estimation. The statistical analysis revealed that the SEA obtained from AR modelling was not sensitive to window duration and sampling frequency. This result verified the consistency of the AR Doppler spectrogram. The white-noise characteristics of the AR modelling error signal indicated that the Doppler blood-flow signal can be adequately modelled as a complex AR process. With appropriate model orders, AR modelling provided better Doppler spectrogram estimates than the periodogram. PMID- 8412378 TI - Effect of method and parameters of spectral analysis on selected indices of simulated Doppler spectra. AB - The sensitivity of Doppler spectral indices (mean frequency, maximum frequency, spectral broadening index and turbulence intensity) to the conditions of spectral analysis (estimation method, data window, smoothing window or model order) increases with decreasing signal bandwidth and growing index complexity. The bias of spectral estimate has a more important effect on these indices than its variance. A too low order, in the case of autoregressive modeling and minimum variance methods, and excessive smoothing, in the case of the FFT method, result in increased errors of Doppler spectral indices. There is a trade-off between the errors resulting from a short data window and those due to insufficient temporal resolution. PMID- 8412379 TI - Analysis of single-unit firing patterns in multi-unit intrafascicular recordings. AB - A system for extracting single-unit activity patterns from multi-unit neural recordings was tested using real and simulated neural data. The system provided reliable estimates of firing frequency for individual units in simulated multi unit data and allowed reliable determinations of the responses of individual cutaneous mechanoreceptor units to 'natural' stimuli such as brushing or pressing on the skin. An implementation of the system, which operated online and in real time, was used to obtain estimates of multiple, single-unit responses from multi unit intrafascicular electrode recordings. The pattern of activity across the population of units in a given recording gave a reliable indication of the type of stimulus that had evoked the activity. It was concluded that this system, used in combination with intrafascicular peripheral nerve recordings, could be used to provide online, real-time information about peripheral stimuli. PMID- 8412380 TI - MicroPACS: a PC-based small PACS implementation. AB - To make picture archiving and communication systems (PACS) a practical reality for small-scale structures, it appears necessary to build a network that integrates and optimises production management, analysis and storage of images, and to include the ability to communicate with other similar remote structures. To solve the various technical problems related to small-scale PACS implementation, MicroPACS, a local area network, was developed around PC-like microcomputers. Every necessary function is assessed and implemented, including a centralised data-base, a distributed image bank, different storage levels and a specific task for image exchanges. An additional protocol was also developed for remote consultation through ISDN. Standardisation of image encapsulation and a point-to-point link between remote sites ensure easy transfers between heterogeneous local networks or bases and integrity of local databases. PMID- 8412381 TI - Nonparametric two-dimensional point spread function estimation for biomedical imaging. AB - The problem of identifying optical system point spread functions (PSFs) arises frequently in the area of image processing and restoration. The paper presents a method for determining two-dimensional PSFs from input/output image signals. The PSF of the system is determined from a set of linear equations involving elements of the input autocorrelation function and the input/output cross-correlation function. The resulting PSF is the one that minimises the sum of squares difference between the actual output image and the predicted one. PMID- 8412382 TI - Laser photoacoustic determination of physiological glucose concentrations in human whole blood. AB - A glucose concentration analysis of human whole blood samples has been accomplished using pulsed laser photoacoustic spectroscopy (LPAS). Using a CO2 laser operating with microJ pulse energies, the technique has shown the required discrimination and sensitivity to determine glucose concentrations within the physiological range (18-450 mg dl-1) in whole blood samples. The sensitivity achieved with this system is comparable to that of the existing commercial enzyme based diagnostic systems presently used in hospital clinical chemistry environments. The technique is compared with other optical methods that have recently been used for glucose determination, and its applicability for use in the development of an in vivo monitor is discussed. PMID- 8412383 TI - Pulse oximetry: theoretical and experimental models. AB - In the paper a pulse oximetry model is developed using an approach which combines both theoretical and empirical modelling. The optical properties of whole blood are measured as a function of cuvette depth by transmission spectrophotometry using red (660 nm) and infra-red (950 nm) light-emitting diodes as light sources. Twersky's theoretical model gives the best fit to the experimental data. A simple theoretical model which takes into account the nonlinear relationship between optical density and cuvette depth is then used to obtain an expression for the R:IR ratio, which relates the measurement of transmission at the two wavelengths. The R:IR ratio is found to be more or less independent of cuvette depth (SD = 0.14 at 100 per cent SaO2). To validate the predictions of the theoretical model, the results of a previous experiment in which the relationship between SaO2 and the R:IR ratio was recorded using a flexible cuvette are used. The experimental values are found to lie within one standard deviation from the theoretical curve relating SaO2 and the R:IR ratio. It is argued that a reasonably accurate model for pulse oximetry which is based on whole blood and not haemoglobin solutions has been developed. PMID- 8412384 TI - Prediction of sampling depth and photon pathlength in laser Doppler flowmetry. AB - Monte Carlo simulation of photon migration in tissue was used to assess the sampling depth, measuring depth and photon pathlength in laser Doppler flowmetry. The median sampling depth and photon pathlength in skin, liver and brain tissue were calculated for different probe geometries. The shallowest median sampling depth found was 68 microns for a 120 microns diameter single fibre probe applied to a one-layered skin tissue model. By using separate transmitting and receiving fibres, the median sampling depth, which amounted to 146 microns for a 250 microns fibre centre separation, can be successively increased to 233 microns when the fibres' centres are separated by 700 microns. Total photon pathlength and thereby the number of multiple Doppler shifts increase with fibre separation, thus favouring the choice of a probe with a small fibre separation when linearity is more important than a large sampling depth. Owing mainly to differences in the tissue g-value and scattering coefficient, the median sampling depth is shallower for liver and deeper for brain, in comparison with skin tissue. For skin tissue, the influence on the sampling depth of a homogeneously distributed blood volume was found to be limited to about 1 per cent per percentage increase in tissue blood content, and may, therefore, be disregarded in most practical situations. Simulations show that the median measuring depth is strongly dependent on the perfusion profile. PMID- 8412385 TI - Self-mixing effect of the semiconductor laser Doppler method for blood flow measurement. PMID- 8412386 TI - Inadequacy of laser Doppler flowmetry in skin areas of the human hand. PMID- 8412387 TI - Impedance plethysmography for the evaluation of pulse-wave velocity in limbs. PMID- 8412388 TI - Extrapolation method to obtain a full projection from a limited one for limited view tomography problems. PMID- 8412389 TI - Hospital drug formularies and use of hospital services. AB - Many hospitals have introduced formularies to reduce hospital pharmacy expense, among other reasons. This study provides empirical evidence of the influence of hospital formulary restrictions on pharmacy charges, all other hospital charges, and on length of stay, using a survey of hospital drug policies and hospital discharge data from Washington State in 1989. Limiting the number of drugs in particular therapeutic categories reduced total charges incurred for gastrointestinal disease and asthma patients, increased total charges for cardiovascular disease patients, and had no effect on charges for infectious diseases patients. Restricting availability of drugs reduced pharmacy charges, but these savings tended to be offset by increases in other charges. Combining the categories, we found that restricting availability of drugs did not affect charges. We conclude that across-the-board restrictions do not result in cost savings, although savings may be realized for particular drug categories. PMID- 8412390 TI - Influence of knowledge and attitudes on the quality of physicians' transfusion practice. AB - Studies evaluating relationships between physician background characteristics (e.g. years of practice and medical training) and quality of care have found few consistent associations. Site of practice variables have been better predictors of quality than have characteristics of individual physicians. This study explores the relationship between physicians' knowledge and attitudes regarding the use of blood products, and the quality of their transfusion practice, based on in-depth physician interviews and medical record reviews. The sample includes 296 transfusion episodes ordered by 17 physicians in 2 teaching hospitals. The physicians had participated in a larger survey of staff surgeons and anesthesiologists in these hospitals. The quality of transfusion practice was defined as the proportion of a physicians' transfusion episodes scored as justified, as determined by explicit chart audit plus implicit physician review. Large baseline differences were observed between the 2 hospitals; 48% of transfusions in hospital A were justified compared with 81% in hospital B. At the physician level, knowledge of transfusion indications and receptivity to input from colleagues were significantly associated with higher quality transfusion practice (standardized betas = 0.41 (P = 0.01), and 0.40 (P = 0.02), respectively). These findings suggest that in addition to organizational context, physician characteristics may be associated with the quality of care related to a specific clinical practice. PMID- 8412391 TI - Competitive bidding for Medicare services. AB - One of the key issues in implementing prospective Medicare fee schedules is how to set prices that accurately reflect competitive market forces. Competitive bidding has long been used in government procurement efforts for nonhealth services. In this paper, we evaluate how provider behavior will be affected if Medicare uses competitive bidding to set Medicare fee schedules. Our model provides several important insights about competitive bidding for health care services. First, the model shows that competitive bidding will lead to 2-stage competition between providers. In the bidding stage, providers will compete to submit winning bids. In the following stage, winning providers will compete for business through marketing efforts that may enhance quality. Second, the model shows how the design of the bidding mechanism affects bidding strategies and the importance of individual provisions within the design, such as penalties for losing bidders. Third, the model demonstrates how competitive bidding will affect quality. It shows how quality may deteriorate if the bidding mechanism chooses an exclusive winner and why naming multiple winners can keep quality at acceptable levels. Finally, we identify criteria for determining whether a particular type of Medicare service is well-suited for competitive bidding. PMID- 8412392 TI - Mental dysfunction and resource use in nursing homes. AB - The role of dementia and other mental disorders in nursing home case-mix classification systems has been an area of controversy. The role of mental dysfunctions was considered in developing a new case-mix measurement system for facility payment in a national demonstration to understand staff time use in nursing homes. Nursing staff (nurses and aides) time and resident assessment data were collected for 6,663 nursing home residents in 6 states. Measures of signs and symptoms of cognitive impairment (dementia), depression, and delirium were created based on items from the new National Minimum Data Set. These measures then were used to determine whether mental dysfunctions were predictive of resource use (nursing staff times and costs) when controlling for other case-mix variables. Cognitive impairment was associated with slightly higher staff time only in less physically-impaired residents without serious medical conditions and not receiving heavy rehabilitation. Similarly, depression and delirium were associated with higher resource use only in selected types of residents. Based on these findings, the new Resource Utilization Groups Version III (RUG-III) contain a major category of residents who are cognitively impaired but not severely dependent in Activities of Daily Living. Depression is used to differentiate subgroups of residents with major medical conditions such as hemiplegia and aphasia. Delirium, when used together with other resident characteristics, was not found useful in explaining resource use. Case-mix groups defined by mental dysfunctions can foster improved care, but careful consideration must be given to appropriate incentives and documentation requirements for providers. PMID- 8412393 TI - Medicare beneficiaries: adverse outcomes after hospitalization for eight procedures. AB - Rehospitalization following surgery is widely recognized as an important outcome measure. The purpose of this study was to identify rehospitalizations for adverse events following 8 procedures, using diagnosis and procedure codes contained in Medicare claims files. Adverse events were broadly defined as: 1) complications; 2) failure of the procedure to achieve its therapeutic goal; and 3) untoward events associated with the natural history of the disease being treated with the procedure. Expert panels identified specific diagnosis and procedure codes that might indicate an adverse event if they appeared on the Medicare record of a rehospitalization. Among patients undergoing percutaneous transluminal coronary angioplasty, almost 36% were rehospitalized for an adverse event within a year of surgery; among patients undergoing coronary artery bypass graft surgery, 20% were rehospitalized for an adverse event. Following the other 6 procedures (cholecystectomy, partial excision of the large intestine, total knee replacement, total hip replacement, replacement of the head of the femur, and reduction of fracture of the femur) between 4% and 9% of patients were rehospitalized for an adverse event. Findings from this exploratory study indicate that rehospitalizations for adverse events appear to be a useful outcome measure for the cardiac procedures; they appear to be less useful for the other procedures, at least at the individual hospital or small area level, because of their relative rarity. Future studies should investigate procedures associated with more frequent rehospitalizations, and medical admissions, which often tend to be associated with higher rehospitalization levels. PMID- 8412394 TI - Relationship between functional status and health-related quality-of-life after myocardial infarction. AB - Despite increasing use in clinical and economic studies, no gold standard exists for the measurement of health-related quality of life (HRQL). One approach to assessing the validity of an HRQL instrument for a particular disease population is to examine the empirical relationship between HRQL patient scores and other accepted measures of health or functional status. In 185 patients (mean age 60 years, 79% male) at six months after myocardial infarction, we examined the relationship between patient responses to the Nottingham Health Profile (NHP), a generic HRQL instrument, and physician classification of patients by two widely used functional status indicators: the New York Heart Association (NYHA) classification and the Karnofsky Performance Status Scale. Analysis of NHP scores by NYHA strata confirms that lower HRQL is associated with poorer cardiac functional status (P < 0.0001) and this gradient is observed over all six NHP domains. Statistically significant (P < 0.001) associations were observed between patients' NYHA class and NHP domain scores for energy (Spearman r = 0.52), physical mobility (r = 0.45) and pain (r = 0.43). NHP scores for patients in NYHA Class I were similar to male population controls. A similarly consistent relationship was found between NHP and Karnofsky. We conclude that the NHP is able to discriminate between patients with differing levels of cardiac functioning as classified by NYHA and patient functioning as classified by Karnofsky. Demonstration of such discriminative properties is one important component in assessing the construct validity of HRQL measures. PMID- 8412395 TI - Ways of knowing: reflections of a health economist. PMID- 8412396 TI - The use of multiple physicians among symptomatic HIV-positive persons. PMID- 8412397 TI - [Thromboangiitis obliterans (Buerger's disease). Study of 41 cases]. AB - BACKGROUND: The aim of this study was to know the prevalence, clinical and immunological characteristics and evolution of thromboangiitis obliterans. METHODS: Between 1982-1990 41 cases of thromboangiitis obliterans were diagnosed from among 373,899 patients registered (11/100,000) according to the clinical and arteriographic criteria. Of these 41 cases 40 were males with mean age 36 +/- 7 years (mean +/- SD) with only two cases being over 45 years of age. In 40 cases was followed for 44 +/- 29 months (mean +/- SD). RESULTS: All the patients had ischemia of the lower limbs, 34% of the upper limbs, 39% superficial thrombophlebitis, 53% Raynaud's phenomenon, 5% mesenteric ischemia, and 7% myocardial infarction. In 30 symptomatic patients anticardiolipin antibodies were determined and one patient was positive for IgG antibodies. In the 23 patients in whom HLA and anticollagen antibody studies were performed a significant increase was found in HLA-B35, HLA-B40 and type VI denaturalized anticollagen antibodies compared to controls. In general the evolution was progressive when the patients continued smoking and favorable when tobacco was given up except in 2 cases who stopped smoking and in whom the disease progressed and death occurred. The total number of patients who died during follow up was 3 (7%), 2 due to mesenteric ischemia and the third during i.v. infusion of PGE1. CONCLUSIONS: The results demonstrate that thromboangiitis obliterans is a rare disease in Spain. The involvement of visceral arteries is not very infrequent with worse prognosis when the mesenteric arteries are involved. There is a significant increase of determined HLA antigens and anticollagen antibodies in the patients with thromboangiitis obliterans. PMID- 8412398 TI - [Vaccination coverage in the districts of the city of Castellon and the relationship with sociodemographic factors: an ecologic study]. AB - BACKGROUND: The state of vaccination in 1987 of those born in the city of Castellon and the relation with a group of sociodemographic variables by districts collected from a study based on the census of inhabitants of 1986 was analyzed. METHODS: The state of vaccination of each child was obtained from the Computerized Municipal Registry of Vaccinations with good vaccination being considered when the child had received all the doses corresponding to the age of two years (four doses of oral trivalent poliomyelitis and three doses of diphtheria-tetanus-pertussis and one dose of diphtheria-tetanus and one dose of triple virus) and was considered as badly vaccinated when one of the doses had been missed. Following classification of each child according to the district of residence, the effect of the state of vaccination, the sex of the child and the month of birth as well as the group of variables collected in the district (ecologic base), the relative risks were estimated by logistic regression analysis. RESULTS: Neither the sex of the child nor the month of birth had any relation with the risk of being badly vaccinated. Upon analysis of the effect being born in a determined district had, five presented between two and four times less risk of bad vaccination than that taken by reference (La Vila) and another three presented up to two times more risk. Among the significant predictors of risk of bad vaccination the following were found: crowding, old age, rate of dependence, rate of widowhood, and the proportion of single people, while the percentage of active women and the density of population were protective. In multivariant analysis, only the proportion of single people and the number of active women persisted as significant predictors. CONCLUSIONS: Although great inequality has been reported in vaccination coverage among the districts of the city of Castellon, the general coverage of the city may be considered as satisfactory in the light of the criteria used and from comparisons with other similar studies. The associations found between bad vaccination and some sociodemographic variables by districts (proportion of single people, and active women) together with possible methodologic limitations, do not allow the important inequalities found to be satisfactorily explained. PMID- 8412399 TI - [thromboangiitis obliterans or Buerger's disease]. PMID- 8412400 TI - [When the physician decides for the patient]. PMID- 8412401 TI - [The consumption of drugs in Spain and its position in European context]. PMID- 8412402 TI - [Large cell anaplastic lymphoma CD30 (ki-1) positive in a patient with human immunodeficiency virus infection]. PMID- 8412403 TI - [Biological markers of alcoholism in alcoholic liver disease]. PMID- 8412404 TI - [Biological markers of alcoholism in alcoholic liver disease]. PMID- 8412405 TI - [Clinical epidemiology of an outbreak of nosocomial infections by Staphylococcus aureus resistant to methicillin and aminoglucosides: efficacy of the measures of control]. PMID- 8412406 TI - Opportunistic/nosocomial infections. Treatment and developmental therapeutics. II. Cryptococcosis. PMID- 8412407 TI - Opportunistic/nosocomial infections. Treatment and developmental therapeutics. Toxoplasmosis. PMID- 8412408 TI - Transdermal drug delivery by passive diffusion and iontophoresis: a review. PMID- 8412409 TI - Steroid hormones and the control of body weight. PMID- 8412410 TI - Multiple regression of skeletal muscle tension on inositol phosphates: cross-talk between signal transduction mechanisms in burn trauma. AB - Skeletal muscle weakness associated with burn trauma prolongs the time of rehabilitation of burn patients. Understanding the underlying chemical changes that impact on physiological tension may provide new therapeutic options for the treatment of burn patients. This report demonstrates the novelty of applying 3 dimensional graphic capabilities, involving area and vector changes to understand variations in inositol derivatives and their co-modulating influence on physiological tension in skeletal muscle. This muscle was distant from the primary anatomical burn site. It was subjected to circulatory shock emanating from burn trauma. Burn injury was achieved by scalding of predefined areas (0, 20% and 50%) on the dorsal and ventral surfaces of mice. At day 21, tension studies via muscle twitch analyses were performed. Through multiple regression, the dependency of physiologic tension was determined with respect to three poly inositol forms each representing independent parameters simultaneously. The contribution of each of these parameters was assigned to a three-dimensional axis. Relationships of tension on three fixed independent parameters were found only for the 20% and 50% burn groups. Vector analysis on a plane in three dimensional space determined the relationship of tension to each of the independent parameters in 20% and 50% burn groups. No significant relationship of tension dependency on three fixed poly-inositol variables was found in the control group. Such vector analysis, using solid and differential analytical geometry, allowed for a clear visualization of the interrelationships that existed between secondary messenger systems (viz, IP3) and a resulting physiologic manifestation (viz, tension). This clear visualization allows for a greater understanding of messenger systems that may lead to more effective treatment of skeletal muscle weakness associated with the systemic effects of severe burn trauma. PMID- 8412411 TI - Adafenoxate abolishes the amnesia induced by neonatal 6-hydroxydopamine treatment in rats. AB - The effect of neonatal 6-hydroxydopamine (6-OHDA) treatment on learning and memory and on the levels of biogenic monoamines in some brain structures, as well as the influence of the nootropic drug adafenoxate on 6-OHDA effect was studied in shuttle box and step down trained rats. In mature rats injected with 6-OHDA postnatal, learning and retention were impaired and the noradrenaline (NA) level in the frontal cortex and hippocampus was decreased. Adafenoxate abolished the amnestic effect of 6-OHDA and restored the NA level to normal in the above mentioned brain structures. This finding suggests the important role of the noradrenergic neurotransmitter system in 6-OHDA-induced amnesia and the favorable effect of adafenoxate on learning and memory impaired by 6-OHDA. PMID- 8412412 TI - Differential effects of calcium channel antagonists in rat normal and skinned fundus. AB - CaCl2 (0.1-25 mM, in K(+)-depolarized tissues), KCl (10-112 mM) and acetylcholine (1 nM-1 mM) produced concentration-dependent contractions of rat isolated fundus. Nifedipine (0.01-500 mcM), diltiazem (0.01-100 mcM) and flunarizine (10-500 mcM) each produced a concentration-related inhibition of the log concentration-effect curve for CaCl2. The rank order of potencies of these antagonists, measured as the IC50 against Ca2+ (25 mM)-induced contraction of depolarized fundus, was nifedipine (1.9 mcM) = diltiazem (2.5 mcM) >> flunarizine (660 mcM). Diltiazem depressed KCl-induced contraction with an effectiveness and potency similar to that displayed against CaCl2 but nifedipine and flunarizine were less effective against contractions to KCl compared to CaCl2. Flunarizine (500 mc), but not the other antagonists tested, depressed Ca2+ (20 mc)-evoked contraction of skinned rat fundus preparations. It is concluded that distinct differences exist between the Ca2+ channel antagonists examined. The action of nifedipine and diltiazem is restricted to the plasmalemma, whereas flunarizine also acts on the intracellular contractile apparatus. PMID- 8412413 TI - Comparative study of different methods of inducing acute and chronic hypercalcemia in rats. AB - Experimental studies are frequently carried out using calcium salt treatment in rats. Modifications in plasma calcium levels could suggest important changes in cellular functions, transmitters and drug responses. Changes during continuous maintenance of hypercalcemia could, on the other hand, be different from those produced by an acute increase in calcemia. Nevertheless, to date no specific studies exist which evaluate and compare the modifications in calcemia and other alterations when different methods of administering acute and chronic calcium are used in rats. This paper presents a method for inducing acute hypercalcemia in Sprague-Dawley rats after intraperitoneal administration of different quantities of CaCl2. Different oral calcium treatments to induce chronic hypercalcemia were also evaluated. Hypercalcemia was more consistent when calcium was administered in both the solid and liquid diets. On day 14 of treatment the highest total and ionic plasma calcium levels appeared in rats fed with CaCO3 in the solid diet (4% Ca) and with CaCl2 in the liquid diet (1.5% Ca). With this treatment hypercalcemia was maintained for 2 months. PMID- 8412414 TI - Clinical-pharmacological study with the two isomers (d-, l-) of fenfluramine and its comparison with chlorpromazine and d-amphetamine: blood levels, EEG mapping and safety evaluation. AB - In a double-blind, placebo-controlled, crossover study, the blood levels and pharmacodynamic properties of single oral doses of 15 and 30 mg d-fenfluramine (d FEN) and 30 mg l-fenfluramine (l-FEN) were investigated as compared with 50 mg chlorpromazine (CPZ) and 20 mg d-amphetamine (AMPH) utilizing pharmaco electroencephalogram (EEG) mapping as well as prolactin and safety assessments. Eighteen healthy young volunteers randomly (Latin square) received at weekly intervals the different treatments. Blood sampling to determine AMPH, FEN and its main metabolite norFEN, as well as plasma prolactic levels, EEG recordings and evaluation of hemodynamics and spontaneous side effects were carried out at 0, 2, 4, 6 and 8 h after drug administration. Before and 24 h after drug intake morning subjective sleep quality was also assessed. Blood level investigations after d FEN demonstrated dose-dependent blood concentrations peaking after 2-4 h and slowly declining thereafter. NorFEN, the main metabolite, exhibited a steady increase in plasma concentrations up to the 8th h. After l-FEN a similar time course was obtained but the concentrations of the parent compound were higher and those of the metabolite lower in comparison with the d-isomer, suggesting stereoselective kinetics. d-FEN induced significant and dose-dependent pharmaco EEG changes characterized by a decrease in total power and combined delta/theta activity and by an increase in beta activity and an acceleration of the centroid of the total activity with no changes in alpha power, which were quite different from those of the reference drugs. l-FEN produced only minimal changes (of the same type as d-FEN). 50 mg CPZ and 20 mg d-AMPH produced significant changes as compared to placebo but with quite opposite patterns, the former exhibiting the typical sedative-neuroleptic profile and the latter that typical of psychostimulants. Dose/treatment- and time-efficacy calculations based on all variables showed in the V-EEG that only 30 mg d-FEN and 50 mg CPZ were different from placebo (with peak effects in the 8th and 4th h, respectively), while in the resting condition (R-EEG) all drugs/dosages but 30 mg l-FEN differed from placebo (with the peak effect occurring in the 4th-6th h after d-FEN and l-FEN, in the 2nd h after CPZ and in the 4th-6th h after AMPH). Plasma prolactin levels were only modified by CPZ (maximal increase in the 2nd h).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412415 TI - Clinical-pharmacological study with the two isomers (d-, l-) of fenfluramine and its comparison with chlorpromazine and d-amphetamine: psychometric and psychophysiological evaluation. AB - In a double-blind, placebo-controlled, crossover study, the single oral dose effects of 15, 30 mg d- and 30 mg l-fenfluramine (FEN) on psychometric and psychophysiological variables were investigated and compared with those of 50 mg chlorpromazine (CPZ) and 20 mg d-amphetamine (AMPH). Eighteen healthy young volunteers randomly (latin square) received at weekly intervals the 6 different treatments. Noopsychic, thymopsychic, psychophysiological and flexibility evaluations were carried out at 0, 2, 4, 6 and 8 h after drug administration. Either dose of d- or l-FEN induced only minimal noopsychic and thymopsychic changes. In contrast, 50 mg CPZ produced highly significant deterioration in attention, concentration, reaction time, psychomotor activity and complex reaction, as well as subjective deactivation, while 20 mg d-AMPH produced subjective activation. Critical flicker frequency was only decreased by 50 mg CPZ and increased by 20 mg d-AMPH. In regard to pupillary measurements, 15, 30 mg d-, 30 mg l-FEN and 20 mg d-AMPH produced opposite changes (widening of pupillary diameter) to those observed after 50 mg CPZ (narrowing of pupil size). Flexibility remained unchanged after FEN treatments, while it consistently decreased after both 50 mg CPZ and 20 mg d-AMPH. Time- and dose/treatment efficacy relations based on discriminant analysis (centroids) of changes in all variables showed 50 mg CPZ as the most psychoactive drug inducing significant changes from the 2nd until the 8th h, followed by 20 mg d-AMPH inducing significant changes from the 4th until the 8th h, but with opposite signs, while FEN treatments exerted only slight effects between the 2nd to 8th h postdrug. PMID- 8412416 TI - [Does examination play any role in continuing education? Experiences from clinical chemistry]. PMID- 8412417 TI - [Improvement instead of suppression of national health services]. PMID- 8412418 TI - [More referrals should go to physicians employed by occupational health services!]. PMID- 8412419 TI - [Influenza time--order the vaccine now!]. PMID- 8412420 TI - [Torture of Palestinians by Israeli physicians must cease]. PMID- 8412421 TI - [Unreasonable charge for the licence]. PMID- 8412422 TI - [Education and research instead of unemployment]. PMID- 8412423 TI - [We are opposing the conscience clause]. PMID- 8412424 TI - [How should osteoporosis be treated?]. PMID- 8412425 TI - [Restenosis after coronary angioplasty. Studies on vascular walls give new hope for prevention]. PMID- 8412426 TI - [Children with Down syndrome. How are new parents supported?]. PMID- 8412427 TI - [A case report. Floating spleen with chronic torsion]. PMID- 8412428 TI - [A complement in the treatment of herniated lumbar disk? Percutaneous nucleotomy- a new principle and technique]. PMID- 8412429 TI - [Homosexuals are completely healthy. But aversion of the society can be a problem]. PMID- 8412430 TI - [Homosexuality is not a disease. Strategy after a complex childhood]. PMID- 8412431 TI - [Medical ethics in time. The threefold time perspective is a source of moral commitment]. PMID- 8412432 TI - [Surgeons as recipients of the Nobel Prize for Medicine. Their life and work in the course of time]. PMID- 8412433 TI - [Development and perspectives of small intestine transplantation]. PMID- 8412434 TI - [Metabolic parameters and neurotensin liberation after resection of the small intestine, syngeneic and allogeneic segment transplantation the rat]. AB - The aim of the following study was to gain some insight into the functional characteristics of different portions of the small intestine after either partial resection or syngeneic and allogeneic transplantation 3 months postoperatively. Nutritional parameters (serum albumin levels, serum triglyceride levels, maltose absorption, excretion of fecal fat) and fat-stimulated neurotensin release were determined in Lewis rats that underwent small-bowel resection (n = 21), syngeneic (Lewis-->Lewis, n = 21), or allogeneic transplantation (Brown Norway-->Lewis, n = 24). The length of the remnant, isograft, or allograft was 27 cm (i.e., one-third of the rat small intestine) and consisted of the proximal (n = 7), middle (n = 7), or distal (n = 7) portion. Three postoperative deaths were due to ileus or pneumonia. After allotransplantation cyclosporine (15 mg/kg body wt. s.c.) was administered for graft acceptance. The control group was not operated upon, but was composed of weight- and age-matched Lewis rats (n = 7). We found that resection of two-thirds of the small intestine led to significantly lower levels of albumin and triglycerides in all three portions investigated (P < 0.01), but did not affect maltose absorption. Excretion of fecal fat was elevated significantly only after distal resection (P < 0.05). When compared to resected animals, syngeneic transplantation did not affect the nutritional parameters, but caused a significantly higher hormone release (P < 0.05) in all three different intestinal grafts. Allogeneic transplantation was successful when the middle or distal portion was grafted. All recipients of proximal allografts showed a severe loss of body weight and died between day 8 and 10 after transplantation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412435 TI - [Comparative experimental studies of mechanical and holmium laser synovectomy]. AB - The goal of this study was to evaluate tissue reactions in rabbit knees following laser synovectomy using a holmium:YAG laser (wavelength: 2.1 microns) and to compare these results with those found after conventional mechanical abrasion treatment. Chronic arthritis was immunologically induced in one knee joint each of 48 rabbits. Twelve served as controls, 12 were sham-operated, 12 were exposed to laser radiation, and 12 others were treated according to conventional methods. In the laser group, a pulse length of 1 ms at a repetition rate of 3 Hz resulted in a pulse energy of 600 mJ. After periods of 1 day, 1 week and 1 and 3 months, respectively, three animals from each group were sacrificed and the synovialis was examined macroscopically and histologically using light and electron microscopy. Edema, acute inflammation, and coagulation necrosis occurred immediately following laser therapy. After 1 week, the synovial layer showed slight fibrosis, which was comparable to that found in the controls. One month later the surface appeared to be smooth. The mechanical abrasion caused hemorrhage and necrosis. Fibrosis was pronounced in all capsular layers, and after 3 months the surface appeared coarse and villous in this group. Based on these preliminary findings, holmium-laser synovectomy may offer an alternative to existing therapeutic techniques in the treatment of rheumatoid arthritis. The arthroscopic approach could prevent additional capsular fibrosis associated with mechanical irritation. PMID- 8412436 TI - [Cystic pancreatic tumors]. AB - Before 1978, where cystic tumors of the pancreas were concerned, pathologists only differentiated between cystic adenomas and cystadenocarcinomas. Recently, however, further tumor entities have been introduced. We now differentiate between the generally benign serous cystic adenoma, the potentially malignant mucinous cystadenoma, the possibly malignant papillary cystic tumor, and the always malignant mucinous cystadenocarcinoma. Other rare tumors include the solid cystic acinous-cell tumor, the cystic islet tumor, and mucinous ductal hyperplasia. Because of their slow growth and primary displacement nature, all of these tumors can usually be detected only after they have attained considerable size. Computed tomography (CT), sonography and endoscopic retrograde cholangiopancreatography (ERCP) have an established role in diagnosis. With these methods, as a rule, it is possible to identify pseudocysts; however, differentiation between the individual tumor types is almost impossible. In our study from 1979 to 1990, we observed ten cases of serous cystic adenomas, nine cystadenocarcinomas, and four malignant papillary-cystic tumors. Of these, nine of the ten serous cystic adenomas, four of the five mucinous cystadenomas, all four papillary-cystic tumors, and five of the nine cystadenocarcinomas were curatively resected. All patients with curatively resected adenomas and one patient with an R1-resected cystic adenoma remained free of recurrence throughout the follow-up period. One 86-year-old female patient in whom a serous cystic adenoma was histologically determined still has no symptoms 8 years after diagnosis despite slow tumor progression. Two of the five patients in whom a cystadenocarcinoma was curatively resected died postoperatively; a third patient died of tumor recurrence 4 months following resection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412437 TI - [Entero-enteral invagination of the small intestine in adults. A rare cause of "uncertain abdomen"]. AB - The present paper reports on a 43-year-old female patient who complained over a number of weeks of paroxysms of crampy pain in the mesogastrium. The diagnosis of ileoileal invagination was only made after she had been admitted to hospital for the third time. The following false diagnoses had been made during the 4-week course of the condition: "pyelonephritis", "acute appendicitis", "chronic appendicitis" and, most recently "psychosomatic abdominal distress". The patient was initially treated with antibiotics and finally with psychotropic drugs. Eventually ultrasound suggested the diagnosis of invagination of the small intestine, which was then verified by conventional barium follow-through radiography. The patient subsequently underwent resection of a segment of the small intestine. Entero-enteric invagination is a very rare event in adults, in which a single (often malignant) cause is identified as triggering the invagination. Peristalsis and ingested food push the tumor distad, thus acting as a motor for invagination. The preferred localizations are the junctions between freely moving segments to retroperitoneally fixed segments (e.g., ileocecal region). Ultrasound of the abdomen is the examination of choice for diagnosis of enteroenteric invagination. Surgical resection of the invagination and its cause (generally tumors) is indicated in adults. PMID- 8412438 TI - [Carotid endarterectomy in the elderly patient. Life table analysis and review of the literature based on 594 consecutive operations]. AB - The significance of age as a prognostic factor was evaluated in patients undergoing carotid artery surgery between 12.8.86 and 31.10.92 in the Department of Vascular Surgery, Erlangen, 594 operations were performed on 546 consecutive patients using somatosensory evoked potentials as routine monitoring. Patients were divided into an older group of 139 (150 operations) with a mean age of 78.2 years and a younger group of 407 (444 operations) with a mean age of 64.4 years. There was no statistical difference in preoperative neurological status and number of risk factors, although there were different distributions of risk factors and concurrent illnesses. Thirty-day mortality rates were 1.7% for younger and 0.7% for older patients, and the incidence of postoperative stroke was 1.7% and 0.7% respectively (not statistically significant). Cumulative 5-year survival was 72.2% and 68.2% respectively. Cardiac diseases were the main cause of death in both groups, mainly fatal cardiac ischemia in younger patients and non-ischemic disease in the older patients. The 5-year stroke-free survival rate was 81% and 91.3% respectively (not significant). There was a trend towards a worse outcome for older patients with stages IIb and IV disease. PMID- 8412439 TI - [Is cholecystectomy a risk factor for colorectal cancer? A meta-analysis]. AB - The number of patients subjected to cholecystectomy has increased since the introduction of laparoscopic methods. Therefore, the question of an association between colorectal cancer (CR-Ca) and cholecystectomy (CHE) is again topical. Several studies have been performed investigating the possibility of a link between cholecystectomy and large bowel cancer. The findings recorded in these studies have been varied and in some cases contradictory. In meta-analyses of the different types of studies (prospective and retrospective) the main question, "Is the risk of colorectal cancer higher after cholecystectomy?" was examined. In prospective matched-pairs studies, 1158 patients who had undergone CHE were compared with 1222 controls. The relative risk (RR) was 1.48, and this result was not significant. The four prospective cohort studies compared the frequency of CR Ca of 22,783 CHE patients with the expected frequency in the population (RR = 0.99). The retrospective studies compared the frequency of previous CHE in 11,797 patients with CR-Ca with the frequency in 33,940 controls without CR-Ca. The calculated odds ratio (O. R.) of the meta-analysis was 1.15, a significant but not clinically relevant increase in risk. Similar results was shown for evaluation of sex difference both in the prospective studies, with an RR of 0.99 for women and 1.00 for men, and in the retrospective studies, with a RR of 1.17 (p < 0.05) for women and 1.09 (n.s.) for men. The results for different location of the tumour show no significant risk differences in prospective studies either for the colon or for the rectum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412440 TI - [Is meta-analysis currently a prerequisite for every relevant literature analysis?]. PMID- 8412441 TI - Increased cytotoxicity of squamous cell carcinoma of the head and neck by combining cisplatin with VP-16 and ciprofloxacin. AB - Chemotherapeutic treatment of squamous cell carcinoma (SCC) of the head and neck has been largely ineffective because of tumor cell resistance. This study examined combinations of cisplatin, 4' demethylepipodophyllotoxin ethylidene D glucoside (VP-16), and ciprofloxacin, a quinolone antibiotic. VP-16 and ciprofloxacin were used in an effort to inhibit DNA repair and increase cytotoxicity. Chemotherapeutic agents often have a direct damaging effect on cellular DNA. Cytotoxicity may be the result of incomplete DNA repair mechanisms; whereas tumor cell resistance to drugs may be due to efficient DNA recovery. A nuclear enzyme especially important to DNA repair and cell growth is topoisomerase II (topo II). Targeted inhibition of topo II by VP-16 and ciprofloxacin may cause increased cisplatin cytotoxicity. SCC cell lines of head and neck origin were treated with a combination of cisplatin, VP-16, and/or ciprofloxacin with cell viability being assessed by the MTT colorimetric assay. Four of five SCC lines examined demonstrated significant augmentation of cisplatin cytotoxicity with the addition of both VP-16 and ciprofloxacin. These in vitro data suggest methods may exist for improving the chemotherapeutic treatment of SCC of the head and neck. PMID- 8412442 TI - DNA- and viral-mediated gene transfer in follicular cells: progress toward gene therapy of the thyroid. AB - The authors investigate the in vitro component of an ex situ strategy for gene transfer into the thyroid gland using DNA complex and retroviral vectors. Canine follicular cells harvested by unilateral lobectomy and grown in low-serum media proliferated in culture and retained their differentiated state as evidenced by morphology and thyroglobulin expression. Transient and "stable" gene transfer in thyroid cells were evaluated by comparing DNA and retroviral transduction techniques. Effective gene transfer and expression was demonstrated by histochemical staining for the marker gene product beta-galactosidase. The efficiency of transduction was assessed using an amphotropic retroviral vector carrying the neomycin resistance gene and semiquantitative polymerase chain reaction (PCR) identification of integrated proviral sequences. This analysis demonstrated a proviral frequency in transduced cultures of 10% to 30%. Transduced cells showed no change in morphology or growth patterns and maintained differentiated function as assessed by antibody staining for thyroglobulin. The thyroid gland is an attractive target for somatic gene therapy because of its large protein-synthetic capacity, sensitivity to hormonal regulation, and proportionately high blood flow. Follicular cell gene therapy may be useful not only for treating congenital or acquired diseases of the thyroid, but also disorders of circulating proteins such as hypopituitarism, hemophilia, and diabetes. PMID- 8412443 TI - Acoustic neuroma volume: MRI-based calculations and clinical implications. AB - This study developed a technique for magnetic resonance imaging (MRI)-based volume calculation and defined the specific volume-diameter relationship in acoustic neuromas (AN). Computer-assisted measurement calculated the area of tumor in each MRI slice. Volume was determined by multiplying area by slice interval. The technique was validated by imaging known volumes of gadolinium. The precision was greater than 95%. Our volume calculations revealed a specific volume-diameter relationship in ANs. The following conclusions can be drawn: 1. our technique can accurately determine AN tumor volume; 2. the AN volume-diameter relationship is a specific function and cannot be predicted by assumptions of tumor shape; 3. the volume-diameter relationship should be combined with clinical data to determine the "break point" at which small increments of diameter produce significant clinical sequelae. PMID- 8412444 TI - Long-term surgical results for congenital aural atresia. AB - Thirty-nine primary surgical cases for correction of congenital aural atresia were reviewed for complications and long-term hearing results. Hearing averages of 25 dB for mild atresia, 40 dB for moderate atresia, and 46 dB for severe atresia were obtained. Serviceable hearing was achieved in 64% of the cases. The two most frequent complications were stenosis and recurrent infections of the cavity and canal skin, with an incidence of 33% and 31%, respectively. Use of split-thickness instead of full-thickness skin graft was associated with fewer complications. The goal of this review is also to share the experience of the senior author in the management of this complex problem. PMID- 8412445 TI - Meniere's syndrome: an atypical presentation of giant cell arteritis (temporal arteritis). AB - The authors discuss three patients with biopsy-proven giant cell arteritis (GCA) who experienced significant auditory and vestibular symptoms. Two of the patients who presented with audiovestibular symptomatology died as a direct result of GCA affecting the vertebral arteries. Postmortem findings are reported. This report illustrates the importance of maintaining a high index of suspicion of GCA in elderly patients who present with the acute onset of audio-vestibular symptoms. These symptoms can be a herald of brain stem infarction if giant cell arteritis is the underlying cause. High-dose prednisone and rheumatologic/neurologic consultation is required. PMID- 8412446 TI - The galvanically-induced vestibulo-ocular reflex in the cat. AB - Rotatory vestibular input is processed by receptor organs and relayed along the vestibular nerves to a central processor, whence the vestibulo-ocular reflex (VOR) is generated. Electrical stimuli applied to the labyrinth can bypass receptors and stimulate the nerve directly, thereby generating horizontal eye movements (EVOR). It was our purpose to mathematically relate the EVOR to the VOR in an effort to generate a parameter by which the experimental effects of ototoxins on the inner ear can be evaluated. A feline preparation was created in which the VOR and EVOR were measured in response to various sinusoidal stimuli. A detailed comparison between VOR and EVOR with respect to gain and phase was performed. From analysis of the data, the differential sensitivity to rotatory amplitude with respect to electrical amplitude was proposed as this parameter. PMID- 8412447 TI - Endoscopic sinus surgery: 4-year follow-up on the first 100 patients. AB - Endoscopic sinus surgery has been reported to be successful, but lack of a standardized classification system hampers comparison of results between studies, and long-term results of surgery have not been reported in a series of consecutive patients. The results of our first 100 endoscopic sinus surgery procedures, reported previously after an average 9-month follow-up, were reviewed with the application of a new classification scheme and in light of a longer (4 year) follow-up. Surgery was successful in all patients whose sinus symptoms resulted from anatomical variations or suppurative infection, but failed in some patients with hyperplastic disease or polyps. In addition, the presence of reactive airway disease or the acetylsalicylic acid (ASA) triad was a bad prognostic sign. The overall success of the procedure in relieving sinus symptoms decreased from 98% at early follow-up to 91% at 4-year follow-up. Sixty-six percent were successful after one procedure and 25% required more than one procedure to achieve success. The decline in success since our first report in 1990 was mostly attributable to late failure in patients with recurrent symptomatic polyposis. Because symptoms may not recur in these patients for up to 3 years, long-term results of surgery for this disorder are necessary. Symptoms of recurrent polyposis can be controlled medically or by revision surgery. PMID- 8412448 TI - Long-term results of mastoid cavities grafted with cultured epithelium prepared from autologus epidermal cells to prevent chronic otorrhea. AB - Chronic otorrhea and recurrent infection from open mastoid cavities are common and troublesome clinical problems for which there is no very satisfactory treatment. The authors have previously described a simple procedure to solve this problem, using autologous cultured keratinocyte layers grafted onto the unepithelialized open mastoid cavities. All procedures are carried out on an outpatient basis without anesthesia, except for local anesthesia for the skin biopsy. Twenty-six patients with 28 "difficult" mastoids, in which otorrhea had been present from 2 to 32 years, have been grafted with a follow-up period varying from 10 to 18 months. Seventeen mastoid cavities became symptom-free as a result of this technique. There were 4 failures, and partial success was shown in 7 cavities as judged by both the patient and by clinical examination. PMID- 8412449 TI - Determining the site of airway collapse in obstructive sleep apnea with airway pressure monitoring. AB - Twenty patients with obstructive sleep apnea (OSA) underwent complete polysomnography and simultaneous upper airway pressure monitoring with a custom made, soft silicone-covered catheter measuring 2.3 mm in diameter. The catheter had four solid-state microtip pressure sensors positioned in the posterior nasopharynx, immediately caudal to the tip of the uvula, at the level of the hyoid bone, and in the midesophagus. The level(s) of airway collapse was determined by changes in the pressure patterns between transducers. In 14 of the 20 patients, airway collapse was confined or initiated at the oropharyngeal region. The obstruction extended to the base of tongue in 7 and to the entire collapsible upper airway in 2 patients. Four patients had collapse at the base of the tongue and 2 had collapse at the hypopharynx. The site of airway collapse remained fairly constant through various sleep stages and positions. Uvulopalatopharyngoplasty (UPPP) and postoperative polysomnography were performed in 4 patients (2 with hypopharyngeal, 1 with base of tongue, and 1 with oropharyngeal airway collapse). Two patients had a favorable response to UPPP. PMID- 8412450 TI - Tongue reconstruction: concepts and practice. AB - Total or subtotal tongue resection results in the potential for severe speech and swallowing disruption and life-threatening aspiration. This report documents the development of a new design for latissimus dorsi flaps used in tongue reconstruction. In order to create a contractile muscle sling which will raise the neotongue toward the palate for speech and swallowing, the flap is harvested with muscle fibers oriented transverse to its long, skin component axis. The flap is then transferred to the oral and oropharyngeal defect and sutured at the level of the mandibular angle to the remaining muscles of mastication. Conventional microvascular anastomosis for free flaps is followed by end-to-end reanastomosis of the hypoglossal nerve stump to the nerve to latissimus dorsi. The skin component is set into the floor of mouth with a curved wedge resected anteriorly, raising a mound to assist with articulation. Fourteen such reconstructions have been performed on patients undergoing glossectomy for cancer. If not invaded by cancer, the glottic larynx was preserved, and the decannulation rate was 80% at a median postoperative interval of 3.2 weeks. Seventy percent of patients achieved oral intake with pureed food or better, and upward motion of the flap was documented by video swallowing studies. Articulation was particularly good. This innervated latissimus dorsi flap design therefore is a viable method for rehabilitation after total or subtotal glossectomy. PMID- 8412451 TI - Nasal mucosal congestion after treatment with bromocriptine. AB - Nine women were given bromocriptine a few days after delivery in order to inhibit lactation. Nasal airway resistance to airflow (NAR) was recorded and blood samples were taken before treatment with bromocriptine, 2 to 3 hours after the first dose of this drug, and after 3 to 5 days on this treatment. All the women had increased nasal congestion after bromocriptine and NAR rose significantly. The prolactin, estradiol, and progesterone hormone levels decreased significantly, but no significant difference was found in the levels of thyroid stimulating hormone (TSH) and vasoactive intestinal polypeptide (VIP). The bromocriptine effect may be caused by different mechanisms. PMID- 8412452 TI - Outcome of sudden deafness with and without vertigo. AB - The outcome of unilateral sudden deafness with and without vertigo was evaluated according to the severity of the initial hearing loss, the shape of the audiogram, and other variables. The subjects for this investigation were 1313 patients who had presented to the authors' university hospital from 1972 to 1990 within 2 weeks of the onset of hearing loss. Thirty percent of the patients had accompanying vertigo. Vertigo occurred frequently in patients with severe hearing loss in the high-tone frequencies. Hearing recovery of high-tone frequencies was worse in patients with vertigo than in those without vertigo even when the initial hearing loss was the same. It is hypothesized that these results can be explained by anatomical factors; the cochlear basal turn being more proximal to the vestibular apparatus than the upper turn. PMID- 8412453 TI - Computer-aided 3-D reconstruction and measurement of the facial canal and facial nerve. I. Cross-sectional area and diameter: preliminary report. AB - Using a computer-aided three-dimensional (3-D) reconstruction method, measurements were made at eight representative sites of diameters and the cross sectional area of the facial canal, facial nerve, and the space between the canal and nerve. Materials used were serial histology sections of seven normal human temporal bones obtained from individuals of different ages. Two areas of decreased cross-sectional area of the facial canal were found: the proximal part of the labyrinthine portion and the middle part of the tympanic portion. These narrowings in the canal appeared to be correlated with decreased superior inferior diameter of the facial canal in those portions, especially in adult temporal bones, and also with decreased cross-sectional area, both of the nerve and of the space between the canal and the nerve, in these portions. The developmental etiology of these narrowings is speculated on, and their possible relationship to Bell's palsy is discussed. PMID- 8412454 TI - Radiologic assessment of the early postoperative total-laryngectomy patient. AB - Pharyngocutaneous fistula is a significant cause of postoperative morbidity following total laryngectomy. The records of 132 patients at the Johns Hopkins Hospital were reviewed retrospectively to determine the role of radiographic contrast studies in the early postoperative period after total laryngectomy. Radiographic studies were performed in 41 cases, of which 38 were cinepharyngoesophagograms. Fistulae occurred postoperatively in 28 patients (21%). In patients with no clinical signs or symptoms suggestive of an impending fistula (fever, wound erythema, wound swelling, or persistent elevated neck drain output), there is no need to perform a cinepharyngoesophagogram before starting oral alimentation. The presence of soft-tissue air in the neck seen on preliminary scout spot films, suggestive of an impending fistula seen in patients who also underwent a neck dissection, did not correlate with a clinical fistula if no extravasation of contrast was observed. PMID- 8412455 TI - Use of the platysma myocutaneous flap for cervical trachea reconstruction: an experimental study in dogs. AB - Cervical trachea reconstruction with a non-stented platysma myocutaneous door flap (NPMCF) was studied in 23 dogs. Window defects involving 5, 10, or 15 rings and the whole anterior tracheal wall were produced in 3 different groups of animals. A segmentary defect involving the resection of 3 tracheal rings but preserving the posterior membranous wall was created in another group. The results were evaluated by clinical follow-up of up to 100 days, by tracheoscopy 1 week after surgery, and by macroscopic and microscopic examination after the animals' natural death or sacrifice. The NPMCF proved to be adequate for the reconstruction of window defects limited to 5 and 10 rings, with success rates of 100% and 75%, respectively. The use of the NPMCF for tracheal reconstruction had the following main advantages: 1. relatively simple, easy, and expeditious surgery; 2. use of a single operative field; 3. availability of large amounts of donor tissue; 4. adequate thickness; 5. reliable irrigation; 6. resistance to environmental exposure as evidenced by absence of infection; and 7. 100% take rate with no granulomas or scar stenosis at the suture lines. However, luminal occlusion due to flap collapse was a 100% fatal complication when the NPMCP was used either for reconstruction of larger window tracheal defects (15 rings-group 3) or segmentary defects (group 4); this was the main limitation of the method, followed by hair growth with accumulation of secretions, which can be easily dealt with. It can be anticipated that this method has a potential for application in well-selected patients. PMID- 8412456 TI - Three-dimensional video imaging in otolaryngology--head and neck surgery. PMID- 8412457 TI - Microsurgical bipolar cautery tonsillectomy. PMID- 8412458 TI - Cutting the middle meatal window. PMID- 8412459 TI - High-risk areas in endoscopic sinus surgery and prevention of complications. PMID- 8412460 TI - Neo-oval window technique and myringo-chorda-vestibulopexy in the BOR syndrome. PMID- 8412461 TI - Harris Mosher, MD--outstanding otolaryngologist. PMID- 8412462 TI - Joseph H. Ogura Memorial Lecture. The vertebrate larynx: adaptations and aberrations. AB - The complex anatomy of the vertebrate larynx shows a steady progression from the simple slit on the floor of the lungfish's pharynx to the fine-tuned mechanism of the human vocal apparatus. The frog's larynx acts as a check valve to prevent collapse of the lungs during a dive, since the animal has no rib cage. The crocodile's laryngeal framework has acquired an epiglottic analogue which fits snugly into the nasopharynx and protects the lower respiratory tract from inundation while the animal drowns its prey. The snake's larynx lies intraorally and can be extended beyond the lower teeth while the reptile leisurely swallows its prey intact. The mammal has acquired a cricothyroid joint, allowing its membranous vocal folds to be stretched during phonation. In Homo sapiens, vocal performance has reached its highest degree of versatility, with a vocal fold capable of adjustment in length, tension and shape. In the course of organic evolution, man appears to have chosen the ability to speak and sing over the security that an intranarial epiglottis would have given him. PMID- 8412463 TI - Directory of otolaryngological societies. PMID- 8412464 TI - [Stomach erosions--classification and diagnosis]. PMID- 8412465 TI - [Spontaneous rupture of an umbilical hernia in decompensated liver cirrhosis]. PMID- 8412466 TI - [Intestinal mycoses--do they exist?]. PMID- 8412467 TI - [Omeprazole and the cytochrome P450 system of the liver]. AB - Enzymes of the cytochrome P450 family play a key role in xenobiotic and thus drug metabolism. The H+,K(+)-ATPase blocker, omeprazole, has been reported to inhibit (subfamily P450IIC) or induce (P450IA) this system. The slower metabolic elimination of diazepam, warfarin und phenytoin is probably due to omeprazole competition for P450IIC; however, this effect was rather negligible in magnitude and not reproducible in each case. Also induction of P450IA was only minor; it resulted in no (theophylline) or trivial changes (caffeine) in elimination of drugs metabolized by this subfamily. Concerns about a possible increase in activation of procarcinogens by P450IA appear illfounded, given the fact that cruciferous vegetables such as broccoli and Brussels sprouts are potent inducers but may rather be associated with a lower incidence of certain types of cancer. PMID- 8412468 TI - [Type of stomach replacement and intestinal motility after total gastrectomy]. AB - 68 disease free patients were investigated prospectively after jejunal gastric replacement [jejunal interposition (JI): n = 40; Roux en Y reconstruction (RY): n = 28]. Used function tests were: scintigraphic emptying studies, hydrogen breath test, hepatobiliary sequential scintigraphy, oral glucose tolerance test, and upper intestinal manometry with 5 measuring ports (times of investigation: 6-12. week and > 1 year postoperatively). Patients with JI showed a better weight gain (p = 0.001), a lower reflux rate (p = 0.01), a slower orocecal transit time (p = 0.05), a prompter bile flow after meal stimulation (p = 0.02), and lower peak glucose levels (p = 0.001). The emptying patterns of the different gastric substitutes were not different. A higher incidence of retrograde contractions in RY and a electromechanical coordination between duodenum and gastric substitute in JI were detected. We conclude that the postoperative function of the jejunal gastric substitute does not depend on it's emptying pattern or volume but on the motility patterns of the entire intestinal tract which seem to be strongly influenced by the choice of the jejunal reconstruction. PMID- 8412469 TI - [The lipid lowering effect of a new guar-pectin fiber mixture in type II diabetic patients with hypercholesterolemia]. AB - The lipid-lowering effect of a new mixture of fiber consisting in guar and apple pectin in combination with apple-pomaces has been estimated in 15 female type 2 diabetics [age mean = 62 (52-70) yr] with hypercholesterolemia (total-chol > 240 mg/dl and LDL-chol > 130 mg/dl). After a dietetic run-in-phase of 3 weeks the patients received the fiber mixture (1 package of 17 g with about 5.9 g water soluble fiber) dissolved in 250 ml water for the next 9 weeks: during the first 3 weeks 2 portions per day, the next 3 weeks twice 1/2 portion and the last 3 weeks one 1/2 portion daily. The fiber mixture had to been consumed 30 minutes before taking a main meal. Regular intake of the fiber product led to a significant decrement in blood lipids, and this improvement under 2 portions daily has been maintained for the most part with the reduced dosage. The total-chol levels decreased by 11.3% during the first 3 weeks of fiber intake (p < 0.05) and by 12.6% during the next 3 weeks (p < 0.05), the decrease during the last 3 weeks under the reduced fiber intake was -9.6% (ns). The HDL-chol levels remained approximately the same during the whole period of observation. The triglyceride concentrations in serum could be lowered by -15.5% during the first 3 weeks of fiber intake, and by -19.2% during the next 3 weeks (p < 0.05), finally the decrease was -12.3% (ns). The atherogenic index (Chol-/HDL-chol) could be reduced on an average of 14.3% in relation to the initial value.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412470 TI - [Duodenal duplication cyst--a rare cause of acute recurrent pancreatitis]. AB - We report on a 42-year-old male patient who has been treated over a period of five years because of acute relapsing pancreatitis. The diagnoses of viral pancreatitis, pancreatic pseudocyst, and choledochal cyst type III (choledochocele) have been erroneously made. The clinical and laboratory findings over this period are presented, compared with one another, and analyzed. The correct diagnosis of a duodenal duplication cyst filled with multiple calculi and lying within the lumen of the duodenum could not be established until intraoperative findings and histological examinations were completed. The pathogenesis of relapsing acute pancreatitis associated with duodenal duplication cyst filled with multiple calculi is proposed, based on the analysis of these findings and the literature review. PMID- 8412471 TI - [Oligosymptomatic sprue--report of 4 cases]. AB - In the last 5 years we observed four cases of oligosymptomatic celiac disease. All patients presented isolated manifestations of malabsorption, but we did not find the typical symptoms of celiac disease, like diarrhea or steatorrhea. Three patients showed an iron deficiency anemia refractory to therapy and one patient a distinct osteomalacia with spontaneous bone fractures. In all these cases we were able to point out the typical microscopic feature and tissue antibodies of the celiac disease. Treating the patients with a gluten-free diet, the symptoms disappeared. These observations demonstrate the importance of the small bowel biopsy in cases of causative unknown isolated manifestations of malabsorption, which possibly belong to the celiac disease. PMID- 8412472 TI - The interaction of etoposide with radiation: variation in cytotoxicity with the sequence of treatment. AB - Etoposide is commonly used in combination with radiation but the best method of integrating these two treatment modalities is not known. In this study, the interaction of etoposide and radiation was investigated using V79 hamster lung fibroblast cells. It was discovered that etoposide (0.25 microgram/ml) was a dramatic radiosensitizer if it was given for a 24 hour exposure after radiation. If the cells were exposed to the same concentration of etoposide for 24 hours prior to radiation, minimal or no radiosensitization occurred. Radiosensitization correlated with a prolongation of the radiation-induced G2 cell cycle arrest. These initial studies indicate that the sequence of radiation and etoposide treatments is important in order to maximize cytotoxicity in V79 cells. Further work using human cell lines will be required to determine the clinical applications of this discovery. PMID- 8412473 TI - Effect of anti-IL-5 monoclonal antibody on allergic bronchial eosinophilia and airway hyperresponsiveness in mice. AB - The effect of pretreatment with rat anti-murine interleukin-5 (IL-5) antibody on antigen-induced bronchial eosinophilia and bronchial reactivity to acetylcholine in mice were studied. Three inhalations of an antigen by actively sensitized animals resulted in an increase in airway reactivity to acetylcholine. Twenty four hours after the final inhalation, the number of leukocytes (mononuclear cells and eosinophils) and the amount of IL-5 in BALF increased significantly. Anti-IL-5 monoclonal antibody inhibited the antigen-induced increase of eosinophils with little effect on bronchial hyperreactivity. PMID- 8412474 TI - Vasopressin and oxytocin but not glucose stimulate hydrolysis of phosphatidylinositol and phosphatidylcholine in a hamster insulinoma. AB - HIT-T15 cells prelabeled with [3H]-arachidonate were incubated for 15 minutes at 37 degrees C in Krebs Ringer buffer (pH 7.1) in the presence and absence of various agonists. Radioactivity remaining in major phospholipids was measured at the end of incubation period. Oxytocin (1 microM), vasopressin (1 microM), and A23187 (5 microM) stimulated loss of radioactivity from phosphatidylinositol and phosphatidylcholine. No loss of radioactivity from either of the phospholipids, however, was detected in the presence of 10 mM D-glucose, an insulin secretagogue in HIT-T15 cells. The lack of phosphatidylinositol response to glucose was also evident when the cells were prelabeled with myo-[3H] inositol. The formation of inositol phosphates at 15 minutes was readily observed upon the treatment of myo [3H] inositol-labeled cells with oxytocin or vasopressin but not glucose or A23187. Inability of glucose to stimulate phosphatidylinositol and phosphatidylcholine hydrolysis in beta cell-derived HIT-T15 cells contrasts sharply with results from studies with pancreatic islets, where hydrolysis of these two phospholipids is readily observed and thought to contribute to the signaling mechanism responsible for stimulation of insulin secretion. PMID- 8412475 TI - Metallothionein induction by cadmium, cytokines, thrombin and endothelin-1 in cultured vascular endothelial cells. AB - Metallothionein induction was investigated using vascular endothelial cells derived from bovine aorta in a culture system. The induction occurred by cadmium (2 and 5 microM) but not by zinc (10 and 300 microM) after a 24-h incubation of the confluent cultures. It was revealed that cytokines including interleukin-1 beta, interleukin-6, tumor necrosis factor alpha and transforming growth factor beta (1 ng/ml each) have a capacity of metallothionein induction. In these inducers, only cadmium and tumor necrosis factor alpha exhibited significant cytotoxicity, suggesting that metallothionein is not induced simply in response to cytotoxicity. It was found that either thrombin or endothelin-1 which are coagulation factor or anti-fibrinolytic factor, respectively, also induced metallothionein synthesis. It was therefore suggested that metallothionein in endothelial cells may be involved in the regulation of the functions of these cells as well as the protection against cytotoxic agents. PMID- 8412476 TI - Influences of regional differences in activities of brush-border membrane peptidases within the rat intestine on site-dependent stability of peptide drugs. AB - Proteolytic hydrolysis rates of neurotensin and acetyl-neurotensin-(8-13) by brush-border membranes from various rat intestinal segments were as follows: jejunum > duodenum approximately jejunoileal junction > ileum > caecum. The rank order of endopeptidase-24.11 activity along the intestine was jejunum > duodenum approximately jejunoileal junction > ileum > caecum. Angiotensin converting enzyme (ACE) had a similar distribution profile as endopeptidase-24.11. Activities of these two enzymes were lower in the distal intestine. Distribution of endopeptidase-2 activity along the intestine was different: ileum > duodenum approximately jejunum approximately jejunoileal junction > caecum. The profiles of differential hydrolysis of neurotensin and acetylneurotensin-(8-13) within the gut corresponded to the distribution of endopeptidase-24.11 and ACE. Moreover, effects of enzyme inhibitors confirm that these two enzymes initiated proteolysis of neurotensin and acetylneurotensin-(8-13). These results suggest that the regional differences in the activities of key brush-border membrane peptidases will affect site-dependent stability of peptide drugs. PMID- 8412477 TI - A novel low-affinity strychnine binding site on renal proximal tubules: role in toxic cell death. AB - Previous studies have shown that the neuronal glycine receptor antagonist, strychnine, mimics the cytoprotective effects of glycine in renal proximal tubules (RPT) (1). The goal of this study was to identify and characterize the site of action of strychnine. 3H-Strychnine bound to RPT in a saturable and reversible manner, and was displaced by unlabelled strychnine (IC50 = 0.87 mM and a Bmax = 57 nmol/mg protein). However, strychnine binding was not inhibited by glycine or related cytoprotective amino acids. Furthermore, the neurotoxicants bicuculline and norharmane, which share the cytoprotective properties of strychnine, inhibited 3H-strychnine binding. These data support the existence of a novel low-affinity strychnine binding site on the RPT plasma membrane that prevents toxic cell death. PMID- 8412478 TI - Cytoprotection by inhibition of chloride channels: the mechanism of action of glycine and strychnine. AB - Previous studies have demonstrated that strychnine mimics the cytoprotective effects of glycine (1) and that strychnine binds specifically to renal proximal tubules (RPT) at cytoprotective concentrations (2). The goal of this study was to determine a mechanism by which strychnine and glycine are cytoprotective. Antimycin A (0.1 microM) caused chloride influx subsequent to mitochondrial inhibition and prior to the release of lactate dehydrogenase (LDH) activity (a marker of cell death/lysis). The addition of strychnine or glycine prevented the chloride influx and LDH release. The chloride channel inhibitors ethacrynic acid, furosemide, anthracene-9-carboxylic acid, DIDS, and SITS decreased LDH release in RPT exposed to antimycin A with a rank order of potency of DIDS > ethacrynic acid = furosemide = anthracene-9-carboxylic acid > SITS. These data, in conjunction with the preceeding paper, indicate a critical role for chloride influx in cell death/lysis; support the existence of a novel strychnine binding site on the plasma membrane of RPT that is coupled to a chloride channel; and suggest that glycine and strychnine are cytoprotective through their inhibition of chloride influx. PMID- 8412479 TI - Effects on open-field behavior of diazepam and buspirone alone and in combination with chronic caffeine. AB - Effects of diazepam (1, 2 mg/kg) and buspirone (1.25, 2.5 mg/kg) on locomotor and rearing activity were observed in rats tested in an open field. Both doses of each drug reduced ambulation. However, for buspirone, this effect was confined to females. Walking and rearing was reduced by the higher dose of diazepam and rearing by both doses of buspirone. In rats that had ingested approximately 26 mg/kg/day of chronic caffeine for seven days prior to and immediately before testing, all effects of diazepam observed earlier failed to achieve significance except for ambulation. However, all earlier buspirone effects (including female only decreased ambulation) were unaffected by the caffeine treatment. It was concluded that buspirone may be preferable to diazepam as an anxiolytic when in the presence of regular caffeine ingestion. PMID- 8412480 TI - Foscarnet inhibits vascular smooth muscle contraction. AB - Foscarnet inhibited noradrenaline and calcium-mediated contractions of the isolated perfused tail artery of the rat. When the noradrenaline contractile response was split into two components, where the first was due to the release of calcium from intracellular stores and the second to the influx of calcium from the extracellular fluid, foscarnet (30 microM) inhibited only the first component of the response. Foscarnet did not inhibit the calcium influx component of the noradrenaline contraction, nor did it affect the inhibition of this component by the L-type calcium channel antagonists verapamil and nicardipine. These results indicate that foscarnet inhibits vascular smooth muscle contraction by inhibiting calcium release from intracellular stores. PMID- 8412481 TI - Electrical stimulation of tooth pulp increases the expression of c-fos in the cat supraoptic nucleus but not in the paraventricular nucleus. AB - Immunoreactivity to Fos protein was detected in the supraoptic (SON) and para ventricular (PVN) nuclei of the cat using immunohistochemical methods. In the intact animal group, only a few Fos-positive neurons were observed in the PVN, but the SON did not contain any positive neurons. Intraperitoneal injection of pentobarbital sodium (Nembutal: 35 mg/kg) induced c-fos expression in the SON, but not in the PVN. Electrical stimulation of tooth pulp with an intensity that was 3 times the threshold of the jaw-opening reflex (200-600 microA) increased the number of Fos-positive neurons in the SON by up to 388% as compared with those of the Nembutal group, whereas the stimulation did not alter the number in the PVN. The increase was observed throughout the extent of the SON. In addition, morphine treatment (2 mg/kg, i. p.), 5 minutes before tooth pulp stimulation, considerably inhibited the increase in the SON. There were no significant differences among the 3 groups (intact, Nembutal, morphine) in the number of positive neurons in the PVN. These findings suggest that these hypothalamic nuclei have different functional roles and that the SON is involved in nociception and/or the consequent emotional and visceral reactions. PMID- 8412482 TI - Acceleration of embryo transport in superovulated adult rats. AB - The etiology of reduced fertility in rodents and humans after exogenous gonadotropin-induced superovulation is unknown. This study examines implantation failure in adult rats induced to superovulate by gonadotropin treatment. After multiple injections of pregnant mare serum gonadotropin (PMSG) followed by human chorionic gonadotropin (hCG), superovulation in adult rats was confirmed on day-1 of pregnancy, however, on day-13, no implantation sites were observed. Daily flushing of oviducts and uteri revealed that the number of embryos recovered from superovulated rats on day-2 was only one-third of that retrieved on day-1. The numbers of embryos retrieved on these days from the oviducts of control rats, however, were almost equivalent. On day-3, no embryos were retrieved from either the oviducts or uteri of superovulated rats, while all embryos from control rats were found in the oviducts. Most embryos recovered from superovulated rats showed normal development. Compared to levels in control rats, serum estradiol (E2) in superovulated rats increased significantly on days-2 and -3 of pregnancy, whereas, serum progesterone (P) levels remained unchanged. Thus, E2/P ratio increased on days-2 and -3 of pregnancy in superovulated adult rats coinciding with the timing of embryo transport acceleration. Our results suggest that superovulated oocytes can be fertilized and reach an early stage of development within the oviduct. Implantation failure in superovulated rats may be due to the accelerated embryo transport resulting from elevated E2/P ratio. PMID- 8412483 TI - An antinociceptive effect of the intraperitoneal injection of nifedipine in rats, measured by tail-flick test. AB - The tail-flick (TF) technique was used to assess the antinociceptive properties of nifedipine (NIF) given intraperitoneally (i.p.). First, the most suitable intensity of the noxious stimulus (temperature of the bulb) has been ascertained and used in the main study. Male Sprague-Dawley rats received NIF, dissolved in dimethyl sulfoxide (DMSO) at the doses of 0.0, 0.5, 2, 5, 10 and 15 mg/kg, or control with no injection. For the main study, the noxious stimulus was limited to 15 sec (cut-off time) and TF latencies were recorded up to 120 min. The antinociceptive response was expressed as the area under the curve for each rat and analyzed by one-way ANOVA. The antinociceptive response to the lower doses of NIF (0.5 and 2 mg/kg) did not differ from control (no injection) and DMSO alone. Significance was found at 5, 10 and 15 mg NIF with no difference among the doses. However, there was an increasing tendency of the mean values from 0.5 to 15 mg NIF resulting in a positive correlation. The correlation coefficient was 0.32483 (p = 0.015) and regression equation Y = (19.37) x dose + 1320. Our data suggest that spinal mechanisms are involved in NIF-induced antinociception. PMID- 8412484 TI - Plasma from patients with the pruritus of cholestasis induces opioid receptor mediated scratching in monkeys. AB - The medullary dorsal horn is a site of action of opiates in producing facial scratching. Extracts of plasma (0.4 microliter) from 4 patients with the pruritus of cholestasis induced facial scratching when microinjected into the medullary dorsal horn of monkeys. This extract-induced scratching could be abolished or prevented by administering the opioid receptor antagonist, naloxone. Neither saline nor an extract of plasma from a nonpruritic cholestatic patient induced scratching when similarly administered. We infer that plasma of patients with the pruritus of cholestasis contains a factor which induces pruritus by a central opioid receptor-mediated mechanism. PMID- 8412485 TI - Characterization of histamine receptors in isolated pig basilar artery by functional and radioligand binding studies. AB - Histamine receptors in pig basilar arteries were investigated in vitro by radioligand binding assays and by measuring the contractile and relaxant responses to histamine. Histamine and 2-pyridylethylamine (H1-agonist) induced concentration-dependent contractions, whereas impromidine (H2-agonist) induced concentration-dependent relaxations. These responses were independent of the presence of endothelial cells. Diphenhydramine (H1-antagonist) partially reversed the histamine-induced contractions to relaxations. Cimetidine (H2-antagonist) potentiated the contraction in a concentration-dependent manner. In the presence of cimetidine, the pEC50 value of histamine for the contraction was 6.30, and diphenhydramine competitively antagonized the histamine-induced contractions (pA2, 7.77). In the presence of diphenhydramine, the pEC50 value of histamine for the relaxation was 5.93, and cimetidine competitively antagonized the histamine induced relaxations (pA2, 6.62). In the binding studies, the Kd value of [3H]mepyramine was 2.1 nM and the Bmax value was 95.6 fmol/mg protein. A competition experiment with diphenhydramine showed that the pKi value (7.51) was similar to the pA2 value. The Kd value for [3H]cimetidine was 126.0 nM and the Bmax value was 459.8 fmol/mg protein. The pKd (6.90) for [3H]cimetidine was similar to the pA2 for cimetidine. The Hill coefficients for these experiments were not significantly different from unity. The present findings indicate that the number of H1-receptors, in terms of the Bmax value for [3H]mepyramine, is smaller than that of H2-receptors, in terms of the Bmax value for [3H]cimetidine. However, the contractile response to histamine is predominantly mediated through stimulation of H1-receptors on vascular smooth muscle cells in pig basilar artery. PMID- 8412486 TI - A comparison of the temporal effects of estradiol and diethylstilbestrol on pituitary content of DNA, prolactin mRNA and prolactin and on serum prolactin levels in ovariectomized Holtzman rats. AB - Ovariectomized Holtzman rats implanted with Silastic capsules of estradiol-17 beta (E2) or diethylstilbestrol (DES) were decapitated at intervals between 4 hours and 10 days of steroid treatment and the anterior pituitaries were collected and analyzed for DNA, prolactin mRNA, and prolactin content. Serum prolactin was also determined. Untreated ovariectomized rats decapitated at 10 days after ovariectomy served as controls. Pituitary DNA levels were not consistently affected by either steroid until 10 days of treatment when levels approximately doubled. In contrast both steroids increased prolactin mRNA and pituitary and serum prolactin levels within 4-8 hours of treatment. Prolactin mRNA levels increased throughout the 10 day period. Pituitary prolactin levels paralleled mRNA levels until day 7 then decreased significantly (P < 0.05) at day 10. Serum prolactin levels increased at 4 and 8 hours, decreased slightly at 24 hours only to increase dramatically at 48 hours. Then at 4 and 7 days of treatment serum levels decreased to the levels seen at 24 hours, followed by a significant (P < 0.05) increase at day 10. Both steroids were equipotent in their effects on serum prolactin; however, DES was significantly (P < 0.05) more potent in increasing pituitary prolactin content than was E2; whereas, E2 was more potent in increasing prolactin mRNA and pituitary DNA. It is concluded that both natural and synthetic estrogens have rapid and sustained, although not equivalent, actions on the levels of prolactin mRNA in the rat. Further, pituitary DNA and pituitary and serum prolactin do not consistently follow the temporal pattern of change seen for prolactin gene expression. PMID- 8412487 TI - Reduction of plasma prolactin by acute administration of CB-154 in ovariectomized F344 and Holtzman rats treated with diethylstilbestrol: a comparison of RIA and Nb2 lymphoma bioassay. AB - Fischer 344 (F344) and Holtzman Sprague-Dawley female rats were ovariectomized and implanted with a s.c. Silastic capsule of diethylstilbestrol (DES). At 1, 4 and 8 weeks of DES exposure, blood samples were obtained by infraorbital sinus puncture under light ether anesthesia before and 2 hours after s.c. administration of CB-154 (2.5 mg/rat). The plasma obtained was assayed for prolactin by radioimmunoassay (RIA) and Nb2 lymphoma cell bioassay (BA). At 1 week of DES treatment plasma prolactin in F344 rats measured by RIA was decreased by CB-154 (approximately 60%) whereas the level measured by BA was not changed and the BA:RIA ratio was increased from 2.4 +/- 0.2 to 4.8 +/- 0.9. CB-154 decreased plasma prolactin levels at 4 weeks but the effect seen by RIA (approximately 80%) was greater than that seen by BA (approximately 60%) and the BA:RIA ratio was increased (2.2 +/- 0.2 vs 3.4 +/- 0.5). By 8 weeks of DES exposure, CB-154 was as effective in reducing the levels measured by BA (approximately 89%) as those measured by RIA (approximately 85%) and the BA: RIA was not affected by the dopamine agonist. In Holtzman rats CB-154 decreased prolactin levels measured by RIA and BA to the same extent at both 1 and 4 weeks resulting in no change in the BA:RIA ratio, but at 8 weeks the BA:RIA was decreased by CB-154 (2.1 +/- 0.1 to 1.7 +/- 0.1). As was observed in F344 rats, the reduction in plasma prolactin induced by CB-154 increased as the duration of DES treatment increased (1 week approximately 45%; 4 weeks approximately 55-60%; 8 weeks approximately 80-85% inhibition) regardless of assay method. It is concluded that DES increases the BA:RIA ratio of plasma prolactin and that acute CB-154 treatment increases, decreases or does not change the BA:RIA ratio depending on the strain of rat used and the duration of DES treatment. PMID- 8412488 TI - Study of in vitro and in vivo stability of liposomes loaded with calcitonin or indium in the gastrointestinal tract. AB - Factors affecting liposome transport to the blood compartment after oral administration to rats were evaluated. A high entrapment of calcitonin (CT) was obtained when the vesicles were prepared by sonication and were composed of egg phosphatidylcholine, cholesterol and stearylamine. In vitro tests showed that the liposomes were stable in light acidic or basic buffers, but that they were partly lysed in pH 2.5, 10 mM bile salts and pancreatin. Oral administration of liposomes entrapping calcitonin in fasting rats showed that the vesicles facilitate transport of the hormone to the general circulation and that they increase the lifetime of 125I-CT in blood. Oral administration of liposomes entrapping radioactive indium in fasting rats did not induce radioactivity in blood. This could be explained by disruption of most of the vesicles in the enterocytes. PMID- 8412489 TI - Effects of Brazilin on glucose oxidation, lipogenesis and therein involved enzymes in adipose tissues from diabetic KK-mice. AB - In order to address the hypoglycemic mechanism of brazilin, effects on glucose metabolism in epididymal adipose tissue from diabetic KK-mice were investigated. Brazilin remarkably lowered non fasting plasma glucose level without any changes in plasma insulin level. Brazilin significantly increased the rate of glucose oxidation and lipogenesis only in the presence of insulin. Activities of glucose 6 phosphate dehydrogenase and fatty acid synthetase, involved in glucose oxidation and lipogenesis respectively, were significantly increased. These results suggest that brazilin might exert hypoglycemic action in insulin resistance state by, at least in part, regulating the enzymatic reaction process involved in glucose metabolism. PMID- 8412490 TI - Developmental changes of testicular activin and FSH receptor mRNA and plasma FSH and inhibin levels in the rat. AB - To investigate the changes in the FSH receptor and the activin receptor during sexual maturation in rat testes, we examined the mRNA levels of the receptors by Northern blot analysis. With a full length rat activin receptor cRNA probe in this study, Northern blot analysis revealed two activin receptor mRNAs (6Kb and 3Kb) in testes. The large messenger (6Kb) was low on day 7, and gradually increased by day 35. On the other hand, the small messenger (3Kb) was low on day 7, began to increase on day 21, and had increased dramatically by day 35. Therefore, compared with our previous data, developmental changes in the mRNA of the activin receptor of male and female gonads showed certain differences. The expression of 3Kb activin receptor mRNA in rat testis may be relevant to the stimulated spermatogenesis in this period. 2.4Kb and 5.5Kb FSH receptor mRNA were revealed on day 7 and decreased to 80% on day 14 and remained at the same level. The Scatchard plots of FSH binding data showed that the binding affinities of testicular FSH were constant at each stage of development. Changes in the mRNA level in FSH receptor were followed by changes in the concentration of FSH receptor in testis. The rise in the plasma FSH level was concomitant with the decrease in the plasma inhibin level by day 14. PMID- 8412491 TI - The late effect of 7-oxo prostacyclin protects the rat heart against calcium paradox. AB - Protective effect of a stable derivative of prostacyclin (7-oxo PGI2) was studied on the model of calcium overload (Ca paradox) 48 h after i.m. administration of the drug in a dosage of 50 micrograms/kg. In isolated rat heart perfused at 37 degrees C and a constant perfusion pressure of 65 mm Hg (Langendorff preparation) Ca paradox was induced by a 3 min perfusion with a calcium-free Krebs-Henseleit (KH) solution and followed by a 10 min perfusion with KH containing normal concentration of calcium. Late protective effect of 7-oxo PGI2 was manifested by: i. improved recovery of heart function (developed pressure and coronary flow) after Ca paradox, ii. better preservation of macroergic phosphates content, iii. better preserved cardiac ultrastructure (sarcolemma) already during Ca depletion phase. One of the proposed mechanisms of the protection afforded by the pretreatment with 7-oxo PGI2 may be that the cell membrane and possibly the intercalated discs become less affected by calcium depletion, resulting in less contracture-mediated membrane damage upon calcium repletion. PMID- 8412492 TI - Postirradiation administration of adenosine monophosphate combined with dipyridamole reduces early cellular damage in mice. AB - The administration of dipyridamole and adenosine 5'-monophosphate (AMP) to mice 5 to 25 min after 1 Gy of total-body gamma irradiation was found to decrease cellular damage, as indicated by the thymidine level in plasma and the amount of saline soluble polynucleotides in the thymus. The drug combination used did not influence similar cytotoxic effects of hydrocortisone. Furthermore, it was shown that the addition of dipyridamole and AMP to in vitro irradiated suspensions of thymocytes enhanced the rejoining processes of DNA strand breaks. Receptor mediated action of extracellular adenosine may be responsible for the therapeutic effects observed. PMID- 8412493 TI - Effects of GABA antagonists on inhibitory avoidance. AB - Experimental data indicate that GABA is involved in memory processes. However there are marked inconsistencies in the reported effects of interference with GABA synaptic activity on memory consolidation of aversively-motivated tasks. Both amnesia and improvement of performance have been reported after treatment with GABA antagonists. These contradictory effects could be explained by procedural differences in training. To test for this possibility rats were trained in passive avoidance using two levels of footshock and injected with a wide range of doses of picrotoxin and bicuculline. Picrotoxin did not modify the conditioned response while bicuculline induced amnesia only with the lower doses at both low and high footshock intensities. It was concluded that GABA is involved in memory consolidation, and that the conflicting results in the literature are indeed due, in part, to procedural differences, and also to the mode of action of these drugs. PMID- 8412494 TI - Molecular determinants of the alpha-2D adrenergic receptor subtype. AB - The alpha-2 adrenergic receptor in the bovine pineal gland and the rodent homologues of the human alpha-2-C10 receptor express alpha-2D subtype pharmacological characteristics. The alpha-2 adrenergic receptor in the chicken pineal expresses characteristics similar to the alpha-2A subtype found in human and pig. The rodent receptors (alpha-2D) contain a serine residue at position 201 whereas the human and porcine receptors (alpha-2A) have a cysteine at this position. Our results indicate that the bovine pineal receptor has a serine at position 201, supporting the alpha-2D classification. However, the chicken pineal receptor also contains a serine at position 201 suggesting that other amino acids may be responsible for the differences in pharmacological characteristics. PMID- 8412495 TI - Subcutaneous injection of an analog of neuropeptide FF precipitates morphine abstinence syndrome. AB - Neuropeptide FF (NPFF) has been shown to exert various antiopiate actions, including precipitation of opiate abstinence syndrome by third ventricle injection in morphine dependent rats. In the present study, dansyl-Pro-Gln-Arg Phe-amide, a lipophilic analog of NPFF, was injected into morphine dependent rats and appropriate sham controls at a dose of 9 mg/kg s.c. Comparison groups were injected with ethanol/water vehicle alone. The NPFF analog precipitated a vigorous opiate abstinence syndrome in morphine dependent rats, but not in sham controls. PMID- 8412496 TI - Studies of the biogenic amine transporters. II. A brief study on the use of [3H]DA-uptake-inhibition to transporter-binding-inhibition ratios for the in vitro evaluation of putative cocaine antagonists. AB - The cocaine receptor on the dopamine transporter is a logical target binding site for the design and synthesis of novel agents for evaluation as possible cocaine antagonists. Although there is no widely accepted and validated assay for detecting a cocaine antagonist, one commonly accepted strategy is to compare the IC50 value of a test agent for inhibition of [3H]dopamine uptake and its IC50 value for inhibition of the binding of a transporter ligand such as [125I]RTI-55. The goal of such a comparison is to guide the synthesis of agents which have high "uptake-to-binding ratios", i.e. agents which are much more potent in the binding assay than they are in the uptake assay. In the present study we tested the hypothesis that ratios different from unity can result from the fact that the two assays are conducted under markedly different conditions. The results showed that conducting the uptake and binding assays under identical conditions reduced the GBR12935 uptake-to-binding ratio of 6.20 (under standard assay conditions) to 0.36. These data indicate that uptake-to-binding ratios must be interpreted with caution, and emphasizes the need for simpler and less expensive methods than cocaine self-administration paradigms to screen compounds as modulators of cocaine reinforcement. PMID- 8412497 TI - Effect of VA-045 on a closed head injury model in rats. AB - The effect of VA-045, a novel apovincaminic acid derivative, was studied in a model of closed head injury (CHI) in rats. CHI was induced by dropping a 400 g weight through a tube from 70 cm above a steel helmet placed on the vertex. Intravenous administration of VA-045 and thyrotropin-releasing hormone (TRH) reduced both the duration of loss of righting reflex and the duration of disruption of spontaneous movement caused by CHI. VA-045, but not TRH attenuated the CHI-induced decreased step through latency (STL) on a passive avoidance task. Over the same time schedule, however, VA-045 did not attenuate the scopolamine induced decreased STL. CHI-induced reduction of the amplitude of somatosensory evoked potentials (SEP) was recovered by VA-045 but not by TRH. These findings indicate that VA-045 may be useful for treating patients with CHI-induced disturbance in consciousness. PMID- 8412498 TI - Involvement of histamine in the control of the waking state. AB - Available evidence indicates that histamine (HA) plays a neuroregulatory role in the waking state. Support for this proposal is provided by electrophysiological, lesion and pharmacological studies, as well as by fluctuations of HA levels according to a circadian pattern. Thus, 1) HA-containing neurons unit activity changes dramatically as a function of behavioral state across the sleep wakefulness continuum, from 2.3 spikes/sec during active waking to virtual silence during slow wave sleep and REM sleep; 2) HA levels reach a minimum during the dark phase followed by an increase during the light period in rats kept under controlled environmental conditions; in addition histidine decarboxylase and HA-N methyl-transferase activities are higher during darkness; 3) lesions or cooling of the posterior hypothalamus in the area where HA-immunoreactive neurons are located gives rise to a state of somnolence or hypersomnia; 4) 2 thiazolylethylamine, the predominantly H1-receptor agonist and thioperamide, the H3-receptor antagonist increase wakefulness in laboratory animals, while the HA synthesis inhibitor a-fluoromethylhistidine, the H1-receptor antagonists mepyramine, diphenhydramine and chlorpheniramine, and the H3-receptor agonist (R) a-Me-histamine produce opposite effects. PMID- 8412499 TI - Meprin activity in rats with experimental renal disease. AB - Activity of renal meprin, a membrane-bound proteinase in the proximal tubule brush border, was measured in normal rats and in two disease groups: chronic puromycin aminonucleoside nephropathy for 12 weeks and streptozocin-induced diabetes for 6 months. Enzyme activity in kidney homogenates was assayed using azocasein as substrate. The mean activity of mephrin was 3.22 +/- 0.34 U/g kidney weight in normal rats. In diabetic animals, enzyme activity was 8.58 +/- 2.11 U/g kidney weight, P < 0.01. In contrast, meprin activity was decreased in rats with puromycin-induced glomerulopathy, 2.13 +/- 0.17 U/g kidney weight, P < 0.01. These findings indicate that meprin activity is elevated in experimental diabetes. Diminished activity of this luminal membrane enzyme in puromycin aminonucleoside nephropathy may contribute to renal injury in this disease model associated with massive urinary protein excretion. PMID- 8412500 TI - Inhibition by N-ethylmaleimide of H(+)-ATPase reduces the cellular action of arginine vasopressin in cultured rat renal papillary collecting tubule cells. AB - We examined whether H(+)-ATPase is involved in the control of cellular action of arginine vasopressin (AVP) to produce adenosine 3', 5'-monophosphate (cAMP) and mobilize cellular free calcium ([Ca2+]i) in rat renal papillary collecting tubule cells in culture. N-Ethylmaleimide (NEM), an inhibitor of H(+)-ATPase, reduced the cellular pH (pHi) dose-dependently. AVP increased cellular cAMP production in a dose-dependent manner. 5 x 10(-6) and 1 x 10(-5) M NEM significantly diminished the AVP-induced increase in cAMP production. 1 x 10(-7) M AVP also increased [Ca2+]i from 111.2 to 189.3 nM, which was significantly reduced by NEM in a dose dependent manner. These results indicate that H(+)-ATPase participates the cellular action of AVP mediated via the pHi control in renal papillary collecting tubule cells. PMID- 8412501 TI - Pancreatic polypeptide stimulates corticosterone secretion by isolated rat adrenocortical cells. AB - Pancreatic polypeptide (PP) dose-dependently enhanced both basal and submaximally ACTH-stimulated corticosterone production by dispersed zona fasciculata/reticularis cells of the rat adrenal gland. Conversely PP did not affect either basal or ACTH- and angiotensin-II-stimulated aldosterone and corticosterone secretion of zona glomerulosa cells. These findings could throw light on the physiological significance of the marked increase in the pancreatic release of PP during stresses. PMID- 8412502 TI - Primary pharmaco-toxicological evaluation of 2-iodomelatonin, a potent melatonin agonist. AB - Series of experiments aimed at a primary pharmaco-toxicological evaluation of 2 iodomelatonin, a high-affinity melatonin analogue, were performed. In the rat ovulation-inhibition model, 2-iodomelatonin was much more potent than either melatonin or 6-chloromelatonin. The acute toxicity was extremely low and close to, though slightly higher than that reported previously for melatonin. In the rat, 2-iodomelatonin was slowly metabolized in vivo; its apparent elimination half-life was about 60 minutes, much longer than that reported for melatonin. The in vitro mutagenesis tests demonstrated clearly that 2-iodomelatonin in concentrations, exceeding the dose range employed in the in vivo studies, was actually devoid of mutagenic effects. The obtained results suggest that 2 iodomelatonin deserves a detailed pharmaco-toxicological evaluation and could be eventually used in pharmacokinetic and pharmacodynamic studies in humans. PMID- 8412503 TI - Alteration of anomeric preference of glucose-induced insulin secretion by glyceraldehyde. AB - Glucokinase activity in pancreatic islets was dose-dependently inactivated by D glyceraldehyde, whereas islet hexokinase activity was not altered. In untreated islets, alpha-D-glucose stimulated insulin secretion more efficiently than beta-D glucose at a glucose concentration of 10 mM. However, glyceraldehyde highly attenuated the insulin-secretory response of pancreatic islets to alpha-D-glucose compared with that to beta-D-glucose. Thus, there was apparently no anomeric preference of glucose-induced insulin secretion in glyceraldehyde-treated islets. Glyceraldehyde affected neither the alpha-preference of glucose phosphorylation by glucokinase nor the beta-preference of glucose phosphorylation by hexokinase. Our study suggests that defective discrimination of glucose anomers by glyceraldehyde-treated islets may be caused by inactivation of glucokinase. PMID- 8412504 TI - Antimicrobial and antioxidant activities of unripe papaya. AB - The meat, seed and pulp of Carica papaya Linn., a popular traditional medicinal herb grown in the tropics, was shown by the agar-cup method to be bacteriostatic against several enteropathogens such as Bacillus subtilis, Enterobacter cloacae, Escherichia coli, Salmonella typhi, Staphylococcus aureus, Proteus vulgaris, Pseudomonas aeruginosa, and Klebsiella pneumoniae. The same parts of papaya were unequivocably demonstrated by electron spin resonance spectrometry to scavenge 1,1-diphenyl-2-picrylhydrazyl (5.8 x 10(14) spins/ml), hydroxyl (5.1 x 10(14) spins/ml) and superoxide (1.2 x 10(14) spins/ml) radicals with the seed giving the highest activity at concentrations (IC50) of 2.1, 10.0 and 8.7 mg/ml, respectively. The superoxide dismutase (SOD)-like activity in the meat, seed and pulp amounts to about 32, 98 and 33 units/ml; comparable to those of soybean paste miso, rice bran and baker's yeast. Vitamin C, malic acid, citric acid and glucose are some of the possible antioxidative components in papaya. Our study correlates the bacteriostatic activity of papaya with its scavenging action on superoxide and hydroxyl radicals which could be part of the cellular metabolism of such enteropathogens. This is indicative of the pathophysiological role of these reactive oxygen species in gastrointestinal diseases and papaya's ability to counteract the oxidative stress. PMID- 8412505 TI - Capsaicin stimulates the migration of human polymorphonuclear cells (PMN) in vitro. AB - Capsaicin, a homovanillic acid derivative in plants, has distinct pharmacological effects in vivo, e.g. it depletes primary afferent neurons of substance P and other tachykinins. The effect of capsaicin on the migration of human neutrophils was tested in concentrations ranging from 10(-8) M to 10(-3) M. In comparison to the control 10(-8) M capsaicin significantly enhanced the migration of PMN cells (CI 1.29; 2P < 0.009) and a peak migration activity was detected with 10(-6) M (CI 1.32; 2P < 0.01). With higher concentrations of capsaicin the CI was not significantly changed. These results show that capsaicin, a plant derived neurotoxin, exhibits a migration modifying activity on human neutrophils through a direct mechanism not mediated by neuropeptides. In addition capsaicin (10(-7) and 10(-5) M) did not affect the luminol-dependent chemiluminescence and therefore does not contribute to a superoxide anion generation in human PMN. PMID- 8412506 TI - Differential effects on GABAA receptor gamma 2-subunit messenger RNA by tolerance to and withdrawal from pentobarbital--an in situ hybridization study. AB - The heterogeneity of the GABAA receptors has been confirmed structurally and functionally. The present study demonstrates the pharmacological heterogeneity of the GABAA receptors. Rats were rendered tolerant to pentobarbital by continuous intracerebroventricular infusion via osmotic minipumps and abruptly withdrawn from pentobarbital. In situ hybridization of mRNA coding for the GABAA receptor gamma 2-subunit showed decreases of mRNA levels in superior and inferior colliculus in pentobarbital tolerant rats compared to rats in withdrawal. In rats 24-hr after withdrawal from pentobarbital, increases of mRNA levels in neocortex, piriform cortex and in granular and Purkinje cell layers of the cerebellum were observed. These results indicate the fast adaptation of GABA synapses in response to abrupt withdrawal from chronic pentobarbital treatment. The differential responsiveness seen in different areas further confirms the pharmacological heterogeneity of the GABAA receptors. The observed increases and decreases of mRNA may underlie, at least in part, the previously reported changes in Bmax of GABAA receptor ligand binding sites. PMID- 8412507 TI - Effects of prior cocaine exposure on subsequent neuronal responsiveness. AB - In vivo electrophysiological experiments were conducted to examine the responses of single, spontaneously-active neostriatal neurons to repeated cocaine exposure. The second of two administrations in a 30 min interval, attenuated the magnitude and duration of cocaine-induced depression in discharge rate, and enhanced the same variables for cocaine-evoked excitation. These findings indicate that the responsiveness of striatal neurons to cocaine may vary predictably as a function of previous exposure. PMID- 8412508 TI - Mutational analysis of muscarinic acetylcholine receptors: structural basis of ligand/receptor/G protein interactions. AB - Molecular cloning studies have revealed the existence of five molecularly distinct muscarinic acetylcholine receptors (m1-m5), which differ in their tissue distribution, ligand binding properties, and functional profiles. Structurally (and functionally), the muscarinic receptors are members of the superfamily of G protein-coupled receptors. A variety of different mutagenesis techniques have been used to study the molecular basis of muscarinic receptor function. This approach has led to the identification of distinct receptor domains (or individual amino acids) predicted to play key roles in ligand binding, agonist dependent receptor activation, and G protein coupling. Since all G protein-linked receptors share a similar molecular architecture, the information gained from the mutational analysis of muscarinic receptors should help delineate functionally important regions of other members of this receptor family. PMID- 8412509 TI - High throughput assay for inhibitors of the epidermal growth factor receptor associated tyrosine kinase. AB - We have developed a colorimetric assay for the examination of inhibitors of epidermal growth factor (EGF) receptor-associated tyrosine kinase in intact cells. EGF receptor from cells treated with inhibitors is captured by an anti-EGF receptor antibody and the phosphotyrosine content is measured by an anti phosphotyrosine antibody. The quantitative assay does not use radioactive substances and is configured for a high-throughput format. Since it is performed in intact cells, substances lower the phosphotyrosine content on the receptor by different mechanisms will be identified. One distinct feature of the assay is that it uses the natural substrate inside the cell as compared to others using artificial substrates in an unphysiological environment. This assay is easy to perform, is reproducible, and is compatible with many organic solvents and tissue culture media. Thus, it is useful for the discovery of EGF receptor kinase inhibitors from natural products or synthetic compounds and is particularly suitable for large-scale screening. PMID- 8412510 TI - Alpha-melanocyte stimulating hormone (MSH) antagonizes the anorexia by corticotropin releasing factor (CRF). AB - We examined a hypothesis that alpha-Melanocyte Stimulating Hormone (MSH) can antagonizes the anorexia induced by intracerebroventricular injection of corticotropin releasing factor (CRF). Food intake of the rats deprived of food for 18 h was significantly inhibited by 4 h after intracerebroventricular injection of CRF. alpha-MSH also suppressed food consumption by 4 h after the treatment. However, simultaneous administration of alpha-MSH attenuated the anorexic effect of CRF at 0-2 h period and food consumption for 4 h was equal to controls. In addition, simultaneous administration of alpha-MSH deleted the stimulatory effects of CRF on serum corticosterone levels. The present experiment demonstrated that alpha-MSH can antagonizes the anorexia and activation of hypothalamo-pituitary-adrenal axis by CRF, suggesting that the processing step of proopiomelanocortin may be important for the induction of anorexia elicited by CRF. PMID- 8412511 TI - Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on plasma and tissue beta endorphin-like immunoreactivity in the most TCDD-susceptible and the most TCDD resistant rat strain. AB - The salient sign of acutely lethal 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) intoxication in rats is hypophagia along with a prominent body weight loss. Endogenous opioid peptides have been implicated as modulators of food intake. In the present study, female rats of both the most TCDD-susceptible (Long-Evans [L E]; LD50 9.8 micrograms/kg) and the most TCDD-resistant strain (Han/Wistar [H/W]; LD50 > 7200 micrograms/kg) were exposed to a single dose of 50 micrograms/kg TCDD ip. This treatment is usually lethal within 1-6 weeks to all L-E rats and nonlethal to all H/W rats. The animals were killed at 1, 4 or 10 days after the treatment. beta-Endorphin-like immunoreactivity (beta-END-LI) was determined by a validated RIA method in the hypothalamus, pituitary, pancreas, duodenum and plasma. TCDD decreased plasma beta-END-LI concentration by 24-37% at every time point of measurement in L-E rats alone. By contrast, feed-restricted controls exhibited an increase of similar magnitude on day 4. Pancreatic beta-END-LI was also elevated in feed-restricted controls at that time point as compared with either the ad libitum control or TCDD group. TCDD appeared to shrink the pituitary gland in both strains by day 4. Pituitary weight was similarly lowered in TCDD-treated rats and feed-restricted controls at the last time point and this reduction was reflected in pituitary beta-END-LI content. Thus, TCDD affects selectively plasma beta-END-LI levels and this impact correlates with its lethality in these strains. PMID- 8412512 TI - Blockade of mu and kappa 1 opioid analgesic tolerance by NPC17742, a novel NMDA antagonist. AB - NPC17742 is a potent competitive NMDA antagonist. Low doses of NPC17742 prevent the development of tolerance to repeated daily injections of the mu agonist morphine and the kappa 1 agonist U50,488H. However, NPC17742 at these same doses is without effect against the kappa 3 analgesic naloxone benzoylhydrazone (NalBzoH). At these doses, NPC17742 does not significantly influence morphine's ED50 value following single or repeated doses of the NMDA antagonist. The ability of NPC17742 to block tolerance to U50488H distinguishes this compound from other NMDA antagonists and raises the possibility of subclasses of NMDA antagonists. Furthermore, these results emphasize the different mechanisms involved with analgesic tolerance among mu, kappa 1 and kappa 3 receptors. PMID- 8412513 TI - A paradoxical stimulatory effect of berberine on guinea-pig ileum contractility: possible involvement of acetylcholine release from the postganglionic parasympathetic nerve and cholinesterase inhibition. AB - The effects of berberine on guinea-pig ileum contractility were studied in both transmurally-stimulated and unstimulated preparations. Transmural stimulation (80 V, 0.5 ms, 0.05 Hz) of the guinea-pig ileal segments produced a twitch response. Berberine (10(-8)-10(-5) M) enhanced this response dose-dependently. Atropine (10(-7) M), but not mecamylamine (10(-5) M), abolished this response. Acetylcholine (3 x 10(-9) and 10(-8) M) also enhanced the response to transmural stimulation. Pretreatment with hemicholinium (3 x 10(-5) M) antagonized the effect of berberine but failed to change that of acetylcholine. Berberine (10(-5) M) also antagonized the alpha 2-adrenoceptor agonist xylazine (10(-8) and 10(-7) M)-induced inhibition of the twitch response to transmural stimulation. In unstimulated ileal preparations, berberine (10(-5) M) produced contractile responses. In these preparations, atropine (10(-6) M), but not mecamylamine (10( 4) M), abolished the response to berberine. Furthermore, berberine (10(-6)-10(-4) M) inhibited dose-dependently the cholinesterase activity of the guinea-pig blood. The results suggest that berberine increases ileal contractility by: 1) increasing acetylcholine release from the postganglionic parasympathetic nerve terminal, 2) increasing acetylcholine retention through an inhibition of cholinesterase activity, and 3) blocking alpha 2-adrenoceptors, possibly in the postganglionic parasympathetic nerve. PMID- 8412514 TI - Visualization of muscarinic m4 mRNA and M4 receptor subtype in rabbit lung. AB - We have used in situ hybridization to study the localization of muscarinic receptor subtype mRNAs and compared this with the distribution of muscarinic receptor subtypes using receptor autoradiography in tissue sections of the rabbit lung. To localize muscarinic receptor subtype mRNAs, subtype-specific synthetic oligonucleotide probes or riboprobes were generated. Two muscarinic receptor mRNAs (m3 and m4) were detected. The m3 mRNA was expressed in high concentration in tracheal smooth muscle, epithelium and blood vessels and to a lesser extent in bronchiolar smooth muscle while the m4 mRNA was abundantly expressed in bronchiolar smooth muscle and alveolar walls. No expression of m1, m2 or m5 mRNAs was found in any lung structures. This was confirmed by Northern blot analysis which revealed only a single transcript of 3.6 kb for m4 mRNA in rabbit lung. With the exception of the airway epithelium and blood vessels, there was a correspondence between mRNA localization and the distribution of muscarinic receptor subtypes visualized by [3H](-)-quinuclidinyl benzilate ([3H]QNB) autoradiographically in the presence of relatively selective muscarinic subtype antagonists (M1-M3) and direct labelling with selective [3H]AF-DX 384. The significance and the reasons for the presence of m4 mRNA and M4 receptors in the rabbit lung are uncertain and warrant further investigation. PMID- 8412515 TI - Automatic quantitative evaluation of autoradiographic band films by computerized image analysis. AB - The present paper describes a new image processing method for automatic quantitative analysis of autoradiographic band films. It was developed in a specific image analysis environment (IBAS 2.0), but the algorithms and methods can be utilized elsewhere. The program is easy to use and presents some particularly useful features for evaluation of autoradiographic band films, such as the choice of whole film or single lane background determination; the possibility of evaluating bands with film scratch artifacts and the quantification in absolute terms or relative to reference values. The method was tested by comparison with laser-scanner densitometric quantifications of the same autoradiograms. The results show the full compatibility of the two methods and demonstrate the reliability and sensitivity of image analysis. The method can be used not only to evaluate autoradiographic band films, but to analyze any type of signal bands on other materials (e.g. electrophoresis gel, chromatographic paper, etc.). PMID- 8412516 TI - Comparison of the cytotoxic effects of cadmium chloride and cadmium metallothionein in LLC-PK1 cells. AB - Recent studies have shown that ionic cadmium (Cd2+) can selectively damage the tight junctions between LLC-PK1 cells. The objective of the present studies was to determine if cadmium that is bound to metallothionein (Cd-Mt) can also damage the junctions between these cells. Cells on Falcon Cell Culture Inserts were exposed to Cd2+ or Cd-Mt from the apical and basolateral compartments. The integrity of cell junctions was assessed by monitoring the transepithelial electrical resistance, and cell viability was evaluated by monitoring the release of lactate dehydrogenase into the medium. Exposure to Cd2+ for 1-4 hours caused a pronounced decrease in the transepithelial resistance without affecting cell viability. By contrast, exposure to Cd-Mt had little effect on the electrical resistance until the cells began to die, which did not occur until 24-48 hours of exposure. Additional results showed that the cells accumulated Cd2+ more rapidly than Cd-Mt. These results indicate that Cd-Mt does not damage the junctions between LLC-PK1 cells, but that it can kill the cells after prolonged exposure. PMID- 8412517 TI - Central nervous system receptor binding profiles of some 2-amino-4-phenyl quinolines: a novel class of alpha 2-adrenoceptor selective ligands. AB - The 2-amino-4-phenyl quinoline moiety is a structural motif common to a number of central nervous system active agents. Extensive radio-ligand receptor binding profiles for several derivatives of this common structural feature have been determined. A high base level of central nervous system receptor affinity was observed with a distinct preference for the alpha-, and in particular the alpha 2 , adrenoceptors. PMID- 8412518 TI - L-NAME and MK-801 attenuate sensitization to the locomotor-stimulating effect of cocaine. AB - Locomotor activity was tested daily following cocaine injections across 21 consecutive days. Subjects were pretreated 30-min before testing with physiological saline, the nitric oxide synthase inhibitor N omega-nitro-L arginine (L-NAME), or the NMDA-receptor antagonist MK-801. Other rats received daily injections of physiological saline instead of cocaine just prior to testing. Rats pretreated with saline and injected daily with cocaine showed increased locomotor activity across the 21-day test period. L-NAME pretreatment depressed cocaine-stimulated locomotor activity, while MK-801 pretreatment increased locomotor activity. To test for behavioral sensitization to cocaine, rats were injected with cocaine 72 hours after their last daily injections. Sensitization was seen in saline pretreated subjects injected daily with cocaine compared to subjects injected daily with saline only, but both L-NAME and MK-801 pretreatment strongly attenuated cocaine sensitization. This finding is consistent with the proposed roles of nitric oxide and NMDA-receptors in cellular adaptation and learning. PMID- 8412519 TI - Improvement in glucose tolerance of insulin resistant rats after chronic or acute administration of benfluorex. AB - The effects of Benfluorex administration on glucose tolerance have been examined in young and old Sprague Dawley rats. The ageing rats were used as a model of insulin resistance. Chronic oral administration of Benfluorex decreased triglycerides levels and normalized glucose tolerance in ageing rats, independently of effects on body weight. Acute intraportal administration of 0.45 mg/kg/h of Benfluorex for 30 min resulted in a 50% increase in glucose tolerance in old rats, but did not modify that in young rats. The improved glucose tolerance brought about by Benfluorex in an animal model of insulin resistance may suggest a wider therapeutic application in man, to include insulin resistant states as type II diabetes or Syndrome X. PMID- 8412520 TI - Serotonin-mediated acute insulin resistance in the perfused rat hindlimb but not in incubated muscle: a role for the vascular system. AB - We have recently shown that the vasoconstrictor serotonin (5-HT) inhibits oxygen uptake in perfused hindlimb possibly due to vascular shunting. Thus in the present study the effect of 5-HT on insulin-mediated glucose uptake was assessed. Rat hindlimbs were perfused at constant flow with medium containing 8.3 mM glucose and a tracer amount of 2-deoxy-D-[1-3]glucose (2DG) with and without 10 microM 5-HT, 15 nM insulin and a combination of the two. 5-HT inhibited insulin mediated stimulation of glucose uptake by 30.4% when added after insulin and 34.4% when added before insulin. In addition, 5-HT inhibited insulin-mediated 2DG uptake by perfused muscles with inhibition ranging from 32% (soleus) to 80% (extensor digitorum longus). The effects of 5-HT on insulin-mediated glucose uptake were partially reversed by vasodilation with carbachol. In contrast to the results for the hindlimb, 10 microM 5-HT had no significant effect on either basal glucose uptake or the stimulation of glucose uptake mediated by 15 nM insulin by isolated incubated soleus or extensor digitorum longus muscles. It is concluded that 5-HT impairs insulin-mediated glucose uptake in the perfused rat hindlimb that may derive from vascular shunting not apparent when muscles are incubated with 5-HT in vitro. These findings may have implications for the link between hypertension and diabetes. PMID- 8412521 TI - Local anaesthetics do not affect protein kinase C function in intact neuroblastoma cells. AB - The effects of local anaesthetics on protein kinase C function in vitro were examined in two model systems: differentiation in mouse Neuro-2a neuroblastoma cells and muscarine M1-receptor mediated phosphoinositide breakdown in human SK-N MC neuroblastoma cells. Staurosporin, a protein kinase C inhibitor, induced marked neuritogenesis in Neuro-2a cells after incubation for 5 h, whereas no effect could be seen after exposure to the local anaesthetics ropivacaine, lidocaine or bupivacaine. In the other model, protein kinase C-mediated regulation of phospholipase C was demonstrated for SK-N-MC cells. Phorbol 12 myristate 13-acetate, a protein kinase C activator, produced a dose-dependent decrease in both basal and carbachol-stimulated phosphoinositide breakdown. Staurosporin blocked this phorbol ester-induced subsensitivity completely, while ropivacaine, lidocaine or bupivacaine did not, suggesting that no functional protein kinase C antagonism is mediated by local anaesthetics. The present study suggests that unlike the reported inhibiting effects of local anaesthetics on purified protein kinase C isoforms, no such modulation is found in intact neuroblastoma cells. PMID- 8412522 TI - Compensation between sympathetic nerves and adrenal medullary activity:effects of adrenodemedullation and chemical sympathectomy on catecholamine turnover. AB - To clarify functional compensation between the sympathetic nerves and the adrenal medulla, the effects of adrenodemedullation and chemical sympathectomy on norepinephrine (NE) turnover in some sympathetically innervated tissues and on NE and epinephrine (E) turnover in the adrenal gland were studied in rats. The rate of NE turnover was significantly accelerated by bilateral adrenodemedullation in the pancreas and in interscapular brown adipose tissue (IBAT). Chemical sympathectomy accelerated E turnover in the adrenal gland. These data indicate that some adrenergic nerves show functional compensation under conditions of depression of the adrenal medulla, and that compensatory acceleration of the adrenal medullary function occurs under conditions of adrenergic dysfunction. PMID- 8412523 TI - Maryland ranks number two in public health. PMID- 8412524 TI - What's up in the management of high-altitude pulmonary edema? AB - Each year, millions of people travel to high-altitude locales. High-altitude pulmonary edema (HAPE), a severe form of acute mountain sickness, is a potentially fatal complication of acute exposure to high altitude. HAPE usually occurs at elevations above 2,500 meters and typically develops in the setting of a rapid ascent to high altitude with inadequate acclimatization and continued physical exertion. Although prompt descent to lower elevation and supplemental oxygen remain the definitive treatment for HAPE, recent investigations indicate that administration of nifedipine or phentolamine may be effective prophylactic or temporizing therapies. PMID- 8412525 TI - Pathogenesis and therapy of pulmonary emphysema. AB - Emphysema is characterized by permanent enlargement of the respiratory air spaces distal to the terminal bronchioles accompanied by destruction of their walls and without obvious fibrosis. We present a review of this disease, discussing its classification, pathology, and therapy. PMID- 8412526 TI - Dipyridamole-thallium stress testing: a local community hospital experience. AB - Clinical data on the first 100 patients who underwent dipyridamole-thallium stress testing in our hospital were reviewed in order to re-evaluate safety guidelines and diagnostic utility in patients with coronary artery disease. Forty patients developed symptoms, including three major ones. One patient had significant bronchospasm, and two others developed significant hypotension with near syncope. The rest had nonspecific chest, shoulder, arm or throat pain, dyspnea, nausea, vomiting, and paresthesia. Most symptoms occurred within the first 10 minutes of dipyridamole infusion. Twenty-eight patients required treatment with intravenous (IV) aminophylline. Of the remaining 60 patients, 30 became hypotensive but remained asymptomatic. Fourteen of 20 patients who underwent coronary angiography had coronary disease. Thirteen were correctly identified by thallium imaging, and only one was identified by electrocardiogram (EKG). Six patients' angiographies showed no evidence of coronary disease. Five of these patients developed perfusion abnormalities during thallium scintigraphy. These results suggest that dipyridamole is a relatively safe drug for pharmacologic stress testing even though the incidence of side effects is relatively high. The high incidence of thallium perfusion abnormalities in patients without coronary disease probably reflects bias in patient selection for coronary angiography, resulting in a relatively small sample of catheterized patients. However, this requires further investigation. PMID- 8412527 TI - Myopathy and cardiomyopathy associated with selenium deficiency: case report, literature review, and hypothesis. AB - A case of selenium-deficiency myopathy, secondary to total parenteral nutrition, is presented. The literature on selenium-deficiency myopathy and cardiomyopathy is reviewed in the context of the literature concerning selenium status in numerous diseases. An hypothesis of a wider role for selenium deficiency in the pathogenesis of myopathy and cardiomyopathy is presented. PMID- 8412528 TI - Vignette of medical history: Toulouse-Lautrec. PMID- 8412529 TI - Imaging case of the month. Parenchymal neurocysticercosis. PMID- 8412530 TI - A look back. Clinical pathological conference. 1953. PMID- 8412531 TI - Public health--the other health care system. PMID- 8412532 TI - Baltimore's public health contributions and today's problems. AB - The health department's early development and the protective roles of public health roles are described. Qualifications for public health leadership are examined. The diminishing and changing nature of services resulting from continuing federal and state cutbacks are discussed. New public health policy requires major changes in national ideology and politics. PMID- 8412533 TI - Baltimore City Health Department: 200 years of progress and partnership. AB - The Baltimore City Health Department began its long history of protecting the health of Baltimore citizens in 1793. An outbreak of yellow fever in Fells Point on the northwest branch of the Patapsco River was the impetus for the governor's appointment of the first two health officers. Since that time, the health department has worked closely with the medical community to promote education and preventive measures (e.g., sewer systems, water chlorination, food inspection) in order to stop the spread of communicable diseases. For 200 years, the Baltimore City Health Department has provided "the advocacy and leadership necessary to ensure the protection and promotion of the health of Baltimore's citizens." PMID- 8412534 TI - Partners in community health: the Baltimore City Health Department, the Johns Hopkins School of Hygiene and Public Health, and the Eastern Health District, 1932-1992. AB - The Eastern Health District of Baltimore was established in 1932 as an early partnership between the Baltimore City Health Department and the Johns Hopkins School of Hygiene and Public Health. Although its heydey occurred in the thirties and forties, for 51 years the district has provided data for researchers studying community health problems; learning opportunities for students in medicine, nursing, and public health administration; and a wide array of clinical and public health services to urban residents. The Eastern Health District concept has been copied worldwide, and although the partnership between the health department and the school of public health has changed over the years, some collaborative activities continue the tradition of research and service provision. PMID- 8412535 TI - Baltimore fluoridates. AB - In all the history of human progress, there is no measure for the advancement or protection of the public health more effective at lesser cost than fluoridation of domestic water supplies for the control of tooth decay, the adjustment of the concentration of a substance naturally present in water to a level found to be optimal for dental health. This is the story of its adoption in a city of nearly one million people. PMID- 8412536 TI - Health indicators of substance abuse problems in Baltimore. AB - Selected health indicators were examined for which Baltimore-specific data were available, including substance abuse treatment admissions, AIDS (acquired immunodeficiency syndrome) case reports, and drug-related emergency room visits and deaths. All indicators reflected high levels and increasing rates of substance abuse problems in Baltimore. Drugs of choice most likely to result in adverse effects were heroin, cocaine, and alcohol--often in combination. The African-American population in Baltimore is disproportionately affected by substance abuse. Emerging drug use patterns are examined, and health service and policy issues are discussed. PMID- 8412537 TI - Geographic distribution of AIDS patients diagnosed at eight Baltimore hospitals. AB - Since the beginning of the AIDS (acquired immunodeficiency syndrome) epidemic in Baltimore, the university hospitals, particularly Johns Hopkins, have diagnosed and treated a disproportionate share of Baltimore patients. Many Baltimore community hospitals are now treating AIDS patients. These hospitals will have an increasing role in providing primary and secondary care as the epidemic expands. PMID- 8412538 TI - Prevalence of human immunodeficiency virus-1 infection in a Baltimore acute care hospital. AB - In conjunction with the Centers for Disease Control and Prevention's (CDC's) national Sentinel Hospital Surveillance System for HIV (human immunodeficiency virus) Infection, we conducted unlinked (anonymous) testing of a systematically selected sample of blood specimens for HIV-1 antibodies to assess the HIV-1 infection level in an urban hospital's catchment population. We excluded specimens from patients with admitting diagnoses often associated with HIV infection. Of the 5,350 specimens tested between April 1988 and October 1989, 106 (2%) were HIV-1 seropositive. HIV-1 seroprevalence in female patients was 1%, and in male patients, 3%. Those 25-44 years of age showed the highest seroprevalence (3.7% in all patients: 6.6% in male patients, 1.1% in female patients). These results demonstrate a high HIV-1 infection level in this patient population and suggest Baltimore hospitals should evaluate the impact and costs of developing routine HIV counseling and testing programs for their patients. PMID- 8412539 TI - Intentional injury--homicide as a public health problem. AB - This article is an epidemiological analysis of Baltimore homicides between 1986 and 1990. The main findings were that young African-American males between 15 and 34 years of age are at greatest risk of homicide victimization. Homicidal events primarily occurred with handguns, in drug-related circumstances, between acquaintances, and in the western police district area. Questions are raised concerning the expansion of the public health professional's role and the use of epidemiologic data for injury prevention programs. This article is the result of a grant from the Centers for Disease Control and Prevention, Division of Injury Control, to the state of Maryland, Department of Health and Mental Hygiene, and the Baltimore City Health Department. PMID- 8412540 TI - Maternal and child health services in the Baltimore City Health Department, 1900 1992. AB - Maternal and Child Health Services in the Baltimore City Health Department, founded in 1934 and 1919, respectively, grew out of concern for maternal and infant death. Increasingly, services have focused on prevention and on building individual health by strengthening the family. The authors identify changing patterns of health care and the health department's continuing role in assuring high quality care and developing guidelines for cost-effective management. PMID- 8412541 TI - The employer and Blue Cross-Blue Shield. 1953. PMID- 8412542 TI - Blue Cross, Blue Shield; "indemnity" or "service". 1953. PMID- 8412543 TI - Selected communicable diseases in Maryland in 1992 (continued). Haemophilus influenzae disease (81). PMID- 8412544 TI - Two cultures revisited: continuously improving in a dichotomous world. PMID- 8412545 TI - Meta-analysis in deriving summary estimates of test performance. PMID- 8412546 TI - Selection of end points in economic evaluations of coronary-heart-disease interventions. AB - Economic evaluations of interventions to lower blood pressure or cholesterol have used different outcome measures, or end points, in the denominator. Some have related the costs of interventions to improvements in physiologic end points such as mm Hg reduction in blood pressure. Some have related costs to avoidance of coronary heart disease (CHD) events or gains in life expectancy. Others have measured improvements in outcome in quality-adjusted life years (QALYs) gained. The different end points imply different analytic perspectives and different data requirements. The more ambitious analyses, though potentially more relevant in certain situations, require more controversial assumptions to be made. This paper illustrates the trade-offs of relevance, accuracy, and precision by reference to an evaluation of drug therapy for hypercholesterolemia undertaken in the United Kingdom. Estimates are given of cost per percentage cholesterol reduction, cost per CHD event avoided, cost per CHD-free year gained, cost per life year gained, and cost per quality-adjusted life year gained. In each case the assumptions required and the potential relevance of the estimate are discussed. The main findings are that: 1) some end points cannot be discounted to present values in a meaningful way and hence the timing of costs and outcomes cannot be reflected in the analysis; 2) the incorporation of quality-of-life adjustments for years on drug therapy and years post-CHD events greatly changes the cost-effectiveness ratios; 3) the rate of discount changes the pretreatment level of cholesterol for which cost per life year gained is equivalent to cost per quality-adjusted life year gained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412547 TI - Confidence bands for receiver operating characteristic curves. AB - Receiver operating characteristic (ROC) curves are mapped out by the two types of errors that are generated by varying the decision threshold used to determine which subjects will be considered abnormal. Under the conventional binormal model for the ROC curve, two-sided and one-sided simultaneous confidence bands for an entire ROC curve, or for a portion of an ROC curve, are constructed by employing Working-Hotelling-type confidence bands for simple linear regression. Pointwise confidence bands are presented for comparison. The cases in which one has only asymptotically normally distributed estimates of the parameters of the ROC curve and consistent estimates of their variance-covariance matrix are emphasized. The methods extend beyond binormal models. PMID- 8412548 TI - Hypothesis generation and the coordination of theory and evidence in novice diagnostic reasoning. AB - This study investigates hypothesis generation and evaluation in clinical problem solving by medical trainees. The study focuses on 1) directionality of reasoning and 2) use of confirmation and disconfirmation strategies in generating and evaluating hypotheses. Two clinical problems were divided into segments of information containing presenting complaint, past history, and physical examination. The initial information indicated a typical myocardial infarct but subsequent information contradicted it. The results showed that the participating students predominantly used forward reasoning and confirmation strategies. When faced with contradictory evidence: 1) second-year students ignored cues in the problem or reinterpreted them to fit the hypothesis; 2) third-year students generated concurrent hypotheses to account for different sets of data; and 3) fourth-year students generated several initial hypotheses and subsequently narrowed the hypothesis space by generating a single coherent diagnostic explanation. The results are discussed in terms of coordination of clinical evidence and its relationship to scientific reasoning. PMID- 8412549 TI - Time preference in medical decision making and cost-effectiveness analysis. AB - Cost-effectiveness analyses usually quantify peoples' attitudes towards delayed outcomes using the exponential discount model. The authors examined three assumptions of this model by assessing the time preferences of individuals towards hypothetical health states and calculating implicit annual discount rates. Of a random sample of medical students, house officers, and attending physicians, 121 participated, reflecting a response rate of 81%. The participants considered three temporary events (colostomy, blindness, depression) that were destined to occur at five sequentially distant times in the future (one day, six months, one year, five years, and ten years). The utility of each prospect was measured using two elicitation techniques (standard gamble and categorical scaling), and 1,394 implicit discount rates were calculated. Of all the discount rates, 62.1% equalled zero, 10.0% were less than 0.00, and 15.7% were greater than 0.10. Mean discount rates for relatively proximal time intervals tended to be larger than those for relatively more distant intervals (0.041 vs. 0.025, p < 0.01). Mean discount rates for blindness tended to be smaller than those for colostomy or depression (0.023 vs. 0.039 vs 0.037, respectively, p < 0.005). Hence, peoples' implicit discount rates are not always small positive numbers that are constant over time and the same for all settings. The authors suggest that the conventional exponential discount model may not fully characterize the time preferences held by individuals. PMID- 8412550 TI - Time-preference studies in the health care context. PMID- 8412551 TI - A comparison of two methods for indexing and retrieval from a full-text medical database. AB - The objective of this study was to compare how well medical professionals are able to retrieve relevant literature references using two computerized literature searching systems that provide automated (non-human) indexing of content. The first program was SAPHIRE, which features concept-based indexing, free-text input of queries, and ranking of retrieved references for relevance. The second program was SWORD, which provides single-word searching using Boolean operators (AND, OR). Sixteen fourth-year medical students participated in the study. The database for searching was six volumes from the 1989 Yearbook series. The queries were ten questions generated on teaching rounds. All subjects searched half the queries with each program. After the searching, each subject was given a questionnaire about prior experience and preferences about the two programs. Recall (proportion of relevant articles retrieved from the database) and precision (proportion of relevant articles in the retrieved set) were measured for each search done by each participant. Mean recall was 57.6% with SAPHIRE; it was 58.6% with SWORD. Precision was 48.1% with SAPHIRE vs 57.6% with SWORD. Each program was rated easier to use than the other by half of the searchers, and preferences were associated with better searching performance for that program. Both systems achieved recall and precision comparable to existing systems and may represent effective alternatives to MEDLINE and other retrieval systems based on human indexing for searching medical literature. PMID- 8412552 TI - Factored stochastic trees: a tool for solving complex temporal medical decision models. AB - The stochastic tree is a continuous-time version of a Markov-cycle tree, useful for constructing and solving medical decision models in which risks of mortality and morbidity may extend over time. Stochastic trees have advantages over Markov cycle trees in graphic display and computational solution. Like the decision tree or Markov-cycle tree, stochastic tree models of complex medical decision problems can be too large for convenient graphic formulation and display. This paper introduces the notion of factoring a large stochastic tree into simpler components, each of which may be easily displayed. It also shows how the rollback solution procedure for unfactored stochastic trees may be conveniently adapted to solve factored trees. These concepts are illustrated using published examples from the medical literature. PMID- 8412553 TI - Inaccuracies in estimates of life expectancies of patients with bronchial cancer in clinical decision making. AB - Approximations of life expectancy in clinical decision making frequently assume constant disease-specific ("excess") mortality hazards over age at diagnosis and over time from diagnosis. This assumption is inconsistent with the longer relative survival of younger patients with bladder cancer and with the declines in mortality hazards from bladder and breast cancers over time from diagnosis. To estimate the error that may result from these assumptions, the authors derived excess mortality hazards from the Surveillance, Epidemiology and End Result (SEER) tumor registry for bronchial cancers stratified by age at diagnosis and time from diagnosis. They compared the life expectancies calculated by a model using an average constant annual cancer-specific mortality hazard over time from diagnosis with those calculated using data-derived cancer-specific annual mortality hazards that varied as a function of time from diagnosis. For younger patients with less advanced disease, the constant-average-mortality model underestimated life expectancies by up to 50% relative to those predicted by the time-variant model. For those over 75 years old at diagnosis, and for all patients with advanced disease, the constant-average-mortality model overestimated life expectancies by up to 65% relative to those predicted by the time-variant model. The authors conclude that predictions of life expectancy with bronchial cancer, and probably with other neoplasms, are limited by the widespread use of oversimplified methods of calculation and by the lack of data describing mortality hazards as a function of time from diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412554 TI - An estimation of life expectancy: the method is a message. PMID- 8412555 TI - Adjusting the number needed to treat: incorporating adjustments for the utility and timing of benefits and harms. AB - The number needed to treat is a unique and cognitively useful summary measurement for the description of medical treatments. However, the original concept lacks the means to account for multiple benefits and harms or differences in the utilities or timings of patient outcomes. The authors describe an "adjusted" number needed to treat that allows for the inclusion of multiple harms and benefits, and also adjustments for the utilities and timings of these outcomes. The expanded version offers a richer description of medical outcomes, and may be utilized as an adjunct to traditional risk-benefit, cost-effectiveness, and decision-analytic techniques. PMID- 8412556 TI - A meta-analytic method for summarizing diagnostic test performances: receiver operating-characteristic-summary point estimates. AB - The authors have devised a meta-analytic method to summarize diagnostic test performances, which they describe along with a clinical example wherein they analyze the performances of real-time ultrasonography in eight studies of the detection of proximal deep venous thrombosis of the lower extremity, selected on the basis of specific inclusion criteria. To evaluate the evidence for fitting a summary receiver operating characteristic (ROC) curve, a test of correlation between the estimates of true-positive rate (TPR) and false-positive rate (FPR) is performed. A high positive correlation argues for summarizing the studies with an ROC curve. In the absence of such correlation, a test of homogeneity is applied separately to the estimates of sensitivity (TPR) and specificity (TNR) to evaluate whether differences among studies are due to chance alone. If the estimates are homogeneous, a summary point estimate and confidence intervals (CIs) are calculated. As a final step, subgroup analyses can be performed to assess alternative explanations of variability in TPR and FPR. Within groups defined by the presence or absence of symptoms of venous thrombosis, a negative correlation between TPR and FPR and homogeneity among studies were found. For symptomatic patients, the summary TPR was 0.97 with a 95% CI of (0.94, 0.99) and the summary TNR was 0.97 (0.95, 0.99). For asymptomatic patients, the summary TPR was 0.66 (0.50, 0.81), and the summary TNR was 0.96 (0.90, 0.99). The difference in TPR between symptomatic and asymptomatic patients was statistically significant. There was no evidence that test referral bias or lack of independent interpretation of test results influenced these findings.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412557 TI - Hemodynamic assessment in managing the critically ill: is physician confidence warranted? AB - Prior to right-heart catheterization of 846 patients, 198 study physicians estimated values of pulmonary capillary wedge pressure (WP), cardiac index (Cl), and systemic vascular resistance index (VRI). The physicians also expressed their confidence in these estimates. Actual values of WP, Cl, and VRI as determined by catheterization enabled the authors to evaluate the quality of the physicians' judgments. The discrimination of the judgments was modest; areas under the ROC curves for WP, Cl, and VRI were 0.724, 0.681, and 0.656, respectively. Calculated using clinically relevant cutoff values, sensitivities were 64%, 50%, and 64%, and specificities were 71%, 75%, and 63%, respectively. Calibration of the estimates of WP, Cl, and VRI was also modest; physicians tended to overestimate low values and underestimate high values. Physicians were generally confident of their estimates, but there was no relation between confidence and accuracy. Experienced physicians were no more accurate than less experienced ones, although they were significantly more confident. The authors conclude that physicians should not use their levels of confidence in their subjective estimates of cardiac function in deciding whether to base therapy on these estimates. PMID- 8412558 TI - Overestimation and the representativeness heuristic. PMID- 8412559 TI - The tissue specific SmN protein does not influence the alternative splicing of endogenous N-Cam and C-SRC RNAs in transfected 3T3 cells. AB - The SmN protein is closely related to the constitutively expressed RNA splicing protein SmB but is expressed only in brain and heart tissue. The inclusion of the VASE exon in the N-Cam mRNA and of the N1 exon in the c-src mRNA correlates with the expression pattern of SmN, being observed in brain and heart but not in other tissues and increasing in amount as SmN levels increase during brain development. However, the artificial expression of SmN in cells in which it is normally absent does not affect the pattern of N-Cam and c-src splicing whilst a cell line lacking detectable SmN is able to include the VASE exon. Hence SmN does not appear to be necessary or sufficient for these tissue-specific and developmentally regulated alternative splicing events. PMID- 8412560 TI - Molecular cloning of rat growth inhibitory factor cDNA and the expression in the central nervous system. AB - Human growth inhibitory factor (GIF) is a new metallothionein-related molecule whose expression is markedly reduced in Alzheimer's disease (AD) brain. We have recently isolated a full-length cDNA for human GIF that has a striking homology to metallothioneins (MTs). As an initial step to better understanding of the biological functions of GIF, we have isolated a full-length cDNA for rat GIF. Comparison of the predicted amino acid sequence with that of human GIF revealed a strikingly high homology (84% identity). Rat GIF cDNA also showed a striking homology to rat MT-1 and MT-II (66% and 62% identities on amino acid sequences, respectively). All cysteine residues were conserved among rat GIF, human GIF and MTs, indicating that cysteine residues play an important function in these molecules. Compared to mammalian MTs, there were 3 bp and 12 bp insertions near the amino-terminal and carboxy-terminal regions in the rat GIF. The rat GIF mRNA was found to be expressed exclusively in the central nervous system, being expressed predominantly in rat cortical astrocytes in primary culture. Although the rat GIF mRNA expression was low during the fetal stage, a dramatic increase of the rat GIF mRNA expression was demonstrated during the postnatal period of 10 17 days. These results indicate that transcriptional regulation of the rat GIF gene is quite different from that of MTs despite its strikingly high homology with MTs. PMID- 8412561 TI - Long-term changes in synthesis of intermediate filament protein, actin and other proteins in pleural sensory neurons of Aplysia produced by an in vitro analogue of sensitization training. AB - Electrical stimulation of peripheral nerves of isolated pleural-pedal ganglia, an in vitro analogue of long-term behavioral training in Aplysia, produced changes in the synthesis of specific proteins in pleural sensory neurons. The changes in incorporation of [35S]methionine into proteins occurring 24 h after electrical stimulation (late) were determined and compared with changes occurring immediately after stimulation (early). Eight proteins were affected 24 h after electrical stimulation. Three of these proteins were also affected immediately after electrical stimulation. Two of the proteins affected late are components of the cytoskeleton. One protein was identified as actin. The other protein was purified from preparative 2D-gels and partial amino acid sequences of 3 peptides derived from this protein were determined. The peptide sequences were found to be identical to those of an Aplysia intermediate filament protein. PMID- 8412562 TI - Rapid response of striatal muscarinic M1-receptor mRNA to muscarinic cholinergic agents in rat brain. AB - The effects of a single administration of muscarinic cholinergic agents on the level of muscarinic M1-receptor messenger RNA (M1-R mRNA) in the rat striatum were studied. Carbachol increased the M1-R mRNA expression rapidly and transiently, while trihexyphenidyl decreased it. These results suggest that muscarinic cholinergic agents participate in the positive regulation of muscarinic receptor mRNA in the early stage after treatment, contrary to the negative regulation in the chronic stage. PMID- 8412563 TI - Cloning and sequencing of a gene encoding the beta polypeptide of Ca2+/calmodulin dependent protein kinase IV and its expression confined to the mature cerebellar granule cells. AB - A cDNA encoding the beta polypeptide of Ca2+/calmodulin-dependent protein kinase IV (CaM kinase IV) was isolated and sequenced from a rat cerebellar cDNA library. By in situ hybridization histochemistry, we demonstrated the differential gene expression for alpha and beta polypeptides of CaM kinase IV in mature and developing rat brains using oligonucleotide probes specific for each polypeptide. PMID- 8412564 TI - Chronic haloperidol does not alter G protein alpha-subunit levels in rats. AB - Female Sprague-Dawley rats were administered continuous haloperidol or no drug for 32 weeks via subcutaneous silastic implants. Three days after drug withdrawal, animals were rapidly decapitated and tissue samples were analyzed for levels of G alpha subunits for Gi and Gs using immunoblotting procedures. No significant differences were seen between groups in the dopaminergic terminal regions of the medial prefrontal cortex, nucleus accumbens, or dorsolateral caudate-putamen. These results suggest that haloperidol administered in a regimen known to produce alterations in several parameters of dopamine function fails to alter the amount of receptor-linked G protein subunits in rat brain. PMID- 8412565 TI - Ontogeny of dopamine transporter mRNA expression in the rat brain. AB - The ontogeny of dopamine transporter (DAT) gene expression was studied in the rat brain. DAT mRNA was first detected in neurons of the ventrocaudal mesencephalon on embryonic day 14 (E14). By E18, intensely expressing neurons in the ventral tegmental area and substantia nigra resembled the pattern found in adult midbrain. DAT mRNA is not abundant in the hypothalamus and the olfactory bulb at any stage of development. These results parallel those noted for tyrosine hydroxylase gene expression in mesencephalon. PMID- 8412566 TI - Molecular cloning and structure of the human (GABATHG) GABA transporter gene. AB - A cDNA molecule encoding the human GABA transporter was synthesized by means of polymerase chain reaction (PCR) technique and used as probe for selecting a human genomic DNA fragment encoding GABA transporter. A positive clone harboring the whole gene was obtained from a human lymphocyte genomic library through utilizing the genomic 'walking' technique. The clone, designated as pHGAT, harbours a DNA fragment of about 39 kb in length inserted into the BamHI site in cosmid pWE15. The gene covers about 25 kb in length and is constituted by four EcoRI restricted fragments which are 13.7 kb, 3.1 kb, 4.2 kb and 7.2 kb long, respectively. The genomic clone contains 15 introns, including two introns prior to the initiator methionine (i.e., the translation start site is in exon 3). Eleven exons encode the twelve transmembrane regions in the transporter protein. Thus as in the case for a number of other membrane proteins, there appears to be a strong tendency for the putative transmembrane domains to be encoded by separate exons. It is noted that the structure of the human GABA transporter gene reported here differs from the mouse gene which is contains 12 introns. PMID- 8412567 TI - Carbachol-induced phosphoinositide metabolism in slices of rat substantia nigra pars reticulata. AB - Carbachol produced an approximately 2-fold stimulation, with respect to basal, of [3H]inositol monophosphate ([3H]IP1) accumulation in microdissected slices of rat substantia nigra pars reticulata (SNr), measured as a percentage of [3H]inositol incorporation. The response to carbachol was inhibited by 75 +/- 7% by 1 microM pirenzepine, consistent with a muscarinic M1 receptor-mediated response. Carbachol-induced [3H]IP1 accumulation was not reduced by pretreatment of the SNr slices with NMDA, in contrast to the 85% inhibition observed in striatal slices. The results suggest that measurement of carbachol-induced phosphoinositide hydrolysis may be a feasible approach for studying muscarinic M1 receptor function in slices of SNr. PMID- 8412568 TI - Changes in preproenkephalin mRNA after unilateral and bilateral labyrinthectomy in the rat medial vestibular nucleus. AB - We examined the expression of preproenkephalin (PPE) mRNA in the vestibular nuclei in rats using in situ hybridization histochemistry. In normal rats, PPE mRNA-positive cells were observed in the medial and spinal vestibular nuclei. After unilateral labyrinthectomy, PPE mRNA was increased in the medial vestibular nucleus on the operated side from the 1st through the 3rd day after the surgery. It is suggested that the changes in PPE mRNA level after labyrinthectomy are involved in vestibular compensation. PMID- 8412569 TI - Chronic ethanol intake decreases vasopressin mRNA content in the rat hypothalamus: a PCR study. AB - Vasopressin mRNA content was studied by quantitative reverse transcriptase polymerase chain reaction (RT-PCR) in the hypothalami of rats chronically treated with ethanol (EtOH). Quantitative RT-PCR allows for the accurate measurement of peptide mRNA levels in discrete regions of the brain of individual animals. EtOH markedly reduced the level of vasopressin mRNA. Furthermore, salt loading was ineffective in inducing a significant increase in vasopressin mRNA level in EtOH treated rats, unlike in controls. The present results suggest that EtOH not only decreases vasopressin mRNA content in the rat hypothalamus, but also impairs its capacity to respond to salt loading. PMID- 8412570 TI - Expression of amyloid precursor protein in a neuronal cell line: functional activity of proximal regulatory elements. AB - To study cell type-specific regulation of the gene for amyloid precursor protein (APP), we have analysed the human APP promoter for transcriptional activity and interaction with nuclear proteins in the neuronal cell line NG108-15. Sequences from -203 to +104 were sufficient for promoter function and regulatory elements within this region (-128 to -63) were identified by DNase I protection experiments. These results suggest that essentially the same proximal elements are recognized by nuclear factors from both neuronal and nonneuronal cells. PMID- 8412571 TI - Regulation of beta-amyloid precursor protein isoform mRNAs by transforming growth factor-beta 1 and interleukin-1 beta in astrocytes. AB - In cultured astrocytes, all three major transcripts of beta-amyloid precursor protein (APP) were expressed with the ratio for APP695, APP751 and APP770 isoform mRNAs being 1:4:2. In comparison with controls, treatment of astrocytes with transforming growth factor-beta 1 (TGF-beta 1) produced about 6 fold increase in total APP mRNA, while elevation in the interleukin-1 beta (IL-1 beta) treated group was small and may relate to the mitogenic effect of IL-1 beta on astrocytes. Treatment of astrocytes with cytokines also produced marked changes in the upregulation in expression of different APP isoforms. The net increase in mRNAs of KPI-containing isoforms APP751 and APP770 was relatively more than for the APP695 isoform. This phenomenon was mainly related to the differences in the expression of KPI-containing APP isoforms and APP695 isoform in the controls. The present findings provide further evidence for the involvement of astrocytes in a cascade of events leading to the development of senile plaques in Alzheimer's disease and Down's syndrome. PMID- 8412572 TI - Effect of d-fenfluramine and 5,7-dihydroxytryptamine on the levels of tryptophan hydroxylase and its mRNA in rat brain. AB - Repeated high doses of d-fenfluramine (dF; 10 mg/kg, i.p. twice daily for 4 days) markedly reduced serotonin (5-HT) concentrations in the hippocampus and striatum of rat brain up to 1 month after treatment, while tryptophan hydroxylase (TPH) levels were reduced only in the hippocampus 5 days after injection. Unlike dF, an intracerebroventricular (i.c.v.) injection of 5,7-dihydroxytryptamine (5,7-DHT 150 micrograms/20 microliters) induced a marked and long-lasting reduction of 5 HT and TPH in both brain regions. Thirty days after injection, 5,7-DHT, but not dF, markedly reduced the number of labelled neurons in the dorsal and ventral regions of the nucleus raphe dorsalis (NRD) and raised the levels of TPH mRNA in the spared neurons at all times examined. TPH mRNA levels were raised 5 and 15 days after dF treatment in the NDR suggesting that changes in the TPH gene expression or transcript stability result following 5-HT depletion. These data are in agreement with the suggestion that 5,7-DHT damages 5-HT nerve terminals and perikarya, but leave unanswered the question of the mechanism of the long lasting reduction of 5-HT levels caused by high, repeated doses of dF. PMID- 8412573 TI - Characteristic localization of non-NMDA type glutamate receptor subunits in the rat pituitary gland. AB - While the involvement of the glutamate receptors in the hypothalamo-hypophyseal system has been clarified at the hypothalamic level, the existence of glutamate receptors in the pituitary gland has remained obscure. We investigated the localization of the glutamate receptors, the non-NMDA type receptor subunits (GluR1-4) in particular, by immunocytochemistry using specific antibodies. The antibodies specific to GluR1, GluR2/3 and GluR4 exhibited the characteristic localization of the receptor molecules in each lobe of the pituitary gland. GluR1 and GluR2/3-positive cells were identified in the anterior and intermediate lobe, and intense terminals of GluR4 and weak terminals of GluR2/3 were observed in the posterior lobe. Such immunoreactivity appeared to be at the axonal terminal of the neurosecretory magnocellular cells. This was confirmed by in situ hybridization histochemistry using specific oligodeoxynucleotide probes and by immunocytochemistry in the neurosecretory magnocellular neurons. The GluR4 mRNA positive signal and GluR4 immunoreactivity were abundantly observed in magnocells of paraventricular and supraoptic nuclei. In addition to the positive fibers, some pituicytes in the posterior lobe exhibited GluR2/3 immunoreactivity. This suggests that pituicytes have non-NMDA type glutamate receptors. Thus, present study suggests that some anterior pituitary cells and pituicytes in the neural lobe are regulated by the glutamate. PMID- 8412574 TI - [Rubidium chloride in the treatment of major depression]. AB - The present study evaluated 20 patients (18 females and 2 males: mean age 55 +/- 8.8 years) suffering from major depression who had been treated with 360/720 mg/die rubidium chloride for 60 days. A gradual and significant improvement in depressive symptoms (HDRS and Zung Scale) and anxiety (Stai X1 and HamARS) was reported. Serum levels were not correlated to clinical improvement. Slight adverse effects were also observed (diarrhea and skin rashes). Rubidium chloride showed a marked and rapid anti-depressive action which was particularly evident in relation to mood, anti-conservative ideas, work, occupational interests and psychomotory slowing-down. It is clear that these symptoms represent the most important aspects of the polymorphous depressive syndrome and, in some ways, this improvement should be interpreted as the effective influence of the drug on the biological contest of mood changes. PMID- 8412575 TI - [Considerations on the concept of suggestion in relation to pharmacology]. AB - After some brief considerations on the history of the concept of suggestion, the authors evaluate the importance of this concept in the doctor-patient relationship in which the pharmacological variable is present. They report their personal experience through a case report and hope for an integrated approach in which the drug could be seen as a biological and relational agent. PMID- 8412576 TI - [The role of the professional nurse in the mental health service]. AB - The role of the nursing staff (which was then not professionally trained) in psychiatric institutes was limited to ensuring that the rules of the institute were respected. After the responsibility for providing assistance had been transferred to Local Psychiatric Services, the role of the professional nurse was radically changed both as a result of the changes in type of patient and the new manner of caring for patients, above all the operational methods used stress the importance of treatment in relations with the patient. The paper analyses the various tasks of the psychiatric nurse which include the initial reception given to the patient, the administration of drugs and evaluation of their effects, the establishment of educational relations with the patient and management of rehabilitation programmes. Whereas the doctor plays a father-role towards the patient due to the authority he possesses, the nurse may be characterised by maternal and caring functions. Given that the lack of theoretical instruments, as possessed by psychiatrists and psychologists, and prolonged contact with patient may expose nursing staff to the risk of damaging their own mental health, it is important to give them specific professional training which will help them handle their relations with psychotic patients. PMID- 8412577 TI - [Integrated therapy in panic attack disorders]. AB - For the past years or so, studies on panic attack disorders (PAD) have accounted for a large proportion of psychiatric research. In spite of the attempts to clarify the etiopathogenesis of PAD, its characteristic psychopathological aspects and the evolutionary stages of its development, its nosographic status is still controversial (despite its inclusion in DSM III-R) and the same is true of the therapeutical approach. Using these observation as their starting point, the authors have divided the present paper into four parts. In the first they attempt to classify PAD in nosographical terms, whereas the second reviews all pharmacological therapies put forward over the past ten years. The third part consists of a short summary of the most widely used psychotherapeutic approaches, and in the last the authors suggest a model of integrated PAD therapy which is still being clinically experimented. The practice of associating pharmacological therapy with a psychotherapeutic approach has certainly been widely used for some time, but the authors underline that the two methods are only fully integrated in the presence of a therapeutic project resulting in a treatment protocol with controls during the course and at the end of treatment. In this context, the psychodiagnostic stage before therapy is particularly important since it provides as precise as possible a picture of the subject's basic personality and psychopathological state. These factors can lead to a wide varation in the choice of drug therapy, and even jeopardise therapeutic success. On the other hand, an exclusively psychotherapeutic approach to PAD does not rule out the onset of recurrent episodes during treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412578 TI - [Psychotherapy, psychoanalysis and the therapeutic process]. AB - The authors examine the transformation undergone by psychoanalytically-based theories in the context of therapeutic application and with regard to the concepts of health or mental disease, and in particular in relation to the consideration of each individuality and intersubjectivity derived from psychotherapeutic relations. The paper discusses works by some of the main supporters of this theory: W. Bion, D. Winnicott and M. Khan in the Freudian field; lastly, with regard to analytical psychology, Jung's idea of the practice of psychotherapy and his aims are fully discussed. The "hermeneutic" aspect of all psychotherapy is then emphasised and it is viewed as the intermingling and reciprocal involvement of three factors: theory, technique and interpersonal relations. PMID- 8412579 TI - [Identification and self-image in adolescents. A comparison between two cultural groups]. AB - Following a review of the literature concerning the problem of identity structure in adolescents, two groups of lower secondary school students were examined, one of which belonged to the Walser community in Valsesia. The family drawing test was used, a personality projective test which, by analysing some indices, enables and understanding to be obtained of the identification processes with parent figures. From the comparison of the two groups it was found that the traditional Walser family offers to adolescents a more clear-cut model and that therefore there is less evident conflict in parent-adolescent relations. In non-Walser families the increased dispersion of data served to create more confused identity processes and greater conflict in intrafamilial affective links. PMID- 8412580 TI - [The psychological impact of HIV infection and the "burn-out" syndrome amongst health care workers dealing with HIV seropositive and AIDS patients]. AB - HIV (Human Immunodeficiency Virus) and AIDS (Acquired Immunodeficiency Syndrome) health care personnel is faced with a life threatening disease and with problems concerning fear of contagion and sense of professional inadequacy in dealing with chronically and terminal patients who may be suffering from psychosocial and neuropsychiatric problems. Therefore, HIV/AIDS health care workers may develop the "burn-out syndrome" (BOS) that is characterized by emotional distress, lowered job productivity and spread of work problems to family and conjugal relationships. BOS aetiology involves individual, organizational and socio cultural factors and its consequences may negatively affect quality of care of HIV/AIDS patients. BOS prevention includes continuing staff training and education on HIV-related issues and support groups for health workers. PMID- 8412581 TI - [From sleep observation to the discovery of trypanosomiasis and glossina]. AB - The african human trypanosomiasis constitutes one of the fascinating subjects of research about the expansion of the European biomedical science. Its history shows the scientific climate in which the investigations took place. Researching the cause which determined it, the researchers got lost in mistakes and comparisons as well as stimulating factors. It is in this atmosphere that trypanosome had been discovered in 1901 and glossina had been confirmed in 1903. But, the disease prevention needed again a lot of researches to correctly understand the complex links between pathogenesis and vectorial transmission in view of effective control and treatment. PMID- 8412582 TI - [Perianal cutaneous ulcerations in the tropical zone. Proposal for a diagnostic and therapeutic decision tree. A case report]. AB - Perianal cutaneous ulcerations in tropical countries have multiple causes: bacterial, viral and parasitic. Taking the opportunity of a case observed in Bambari (CRA) of which the etiologic origin was not clearly established, the authors offer a proposal for a diagnostic and therapeutic protocol aimed at easing decision. Etiologies quickly cured, such as amoebiasis and tuberculosis have to be explored in priority. PMID- 8412583 TI - [Postoperative nosocomial infections in the Cameroon hospital milieu. Epidemiology and prevention]. AB - The paper aims at describing easy steps: asepsis and hygiene to prevent nosocomial infections within surgery unit in Hospitals with limited budget. Essential and compulsory they are: cleansing of the surgery unit before any disinfection, behaviour of the staff in relation to aerobic contamination, maintenance of equipment, prophylaxis applied to patients antibio-prophylaxis is also mentioned complementary to that steps. Possible strategies of fight and control are mentioned, including awareness and full participation of all. PMID- 8412584 TI - [Study of hypertension in the Ivory Coast urban milieu]. AB - The authors carried out a sample survey in the town of Abidjan to rate high blood pressure in adult. They evaluate the prevalence rate of high blood-pressure at 13.4%. In this study one may consider as a risk factor: age, obesity, illiteracy, either in isolation and as a matter of course the three together. The role of age as confounding factor appears essential in pluri-factorial analysis and has to be systematically taken into account at the occasion of further studies. PMID- 8412585 TI - [The human immunodeficiency virus (HIV) in Australian Aborigines of Queensland and Torres Strait Island]. AB - The authors report on 29 notifications of Human Immunodeficiency Virus infection in Aborigines and Torres Strait Islanders in Queensland from 1 July 1985 to 31 August 1991. Despite previously expressed concerns regarding the potential for a Pattern II (heterosexual) epidemic, based on reported data for other Sexually Transmitted Diseases, there have been no published reports confirming HIV infection in the Aboriginal and Torres Strait Islander population prior to this study. Although the prevalence of diagnosed HIV infection is comparable to that of the remaining population, there are early indications of differences in the pattern of infection and transmission. PMID- 8412586 TI - [Ankylosing spondylitis in Togolese patients]. AB - A transversal study was carried out to determine the frequency and the semiology of ankylosing spondylitis in outpatient rheumatology department at Lome (Togo). 13 of 2.626 examined patients in three years suffered of ankylosing spondylitis. All patients were male and a mean age of 23 at the appearance of the disease. Any had family case history but all presented a pelvis--rachis lesion. Three of them got a peripheral articular attack and 5 an enthesitis. Extra articular attack in 2 patients came out in the form of aqueous diarrhoea. Any patient had previous uveitis nor psoriasis, nor chronic enterocolopathy. All patients suffered from sacro-ileitis. Two patients got, in addition, some syndesmophytes. A bilateral coxitis made the disease more difficult to be treated in a patient whose ankylosing spondylitis was evoluting for 5 years. The symptomatology observed was comparable to the one in Southern Africa. It was like the one in Black Americans not carriers of HLA B27. Some further studies will make more precis the epidemiological and genetical profile of ankylosing spondylitis in Togo. PMID- 8412587 TI - [Decrease in chloroquine resistant Plasmodium falciparum in the Abidjan region (Ivory Coast)]. AB - The authors reviewed the evolution of Plasmodium falciparum chloroquine resistance in greater Abidjan for the year 1990. They used the classical course of treatment consisting of 25 mg per kg of bodyweight distributed on three days. Parasitaemia and body temperature controls were carried out on day O, day 2 and day 7. The study, which involved 547 children averaging 38.3 months of age disclosed an early in vivo resistance in the range of 18.20% as against 29.6% in 1988. PMID- 8412588 TI - [Short term projections of infection by the human immunodeficiency (HIV) and the acquired immunodeficiency syndrome (AIDS) in Cameroon]. AB - As of December 1991, Cameroon has reported 827 cases of AIDS. The results from the Sentinel Surveillance System show a seroprevalence of 1.3% HIV1 among pregnant women, 2.5% in people attending STD clinic and 3.5% in tuberculosis patients. The World Health Organization projection model was used to make a short term projection of HIV infection and AIDS cases. Results show that the number estimated of HIV infected populations varies between 24 to 45,000 people by the year 1995. Results show also an estimated 8,500 cumulative AIDS cases. Even in a low prevalence country as Cameroon the impact of the HIV epidemic is important and will result in a burden for the health care system. PMID- 8412589 TI - [Prenatal surveillance and determinants of prenatal follow-up in the Monastir health district]. AB - Prenatal monitoring is a prime strategy to improve the end of pregnancy. To estimate the prevalence rate and the conditions determining such a monitoring, the authors carried out a survey on 1190 pregnant women in the Monastir district (Tunisia). The rate of satisfactory monitoring (4 check-up and more) is 45.8 p.c. Age and high parity have a negative influence, but instruction, occupation and a bad out come of the last pregnancy favour a satisfactory prenatal monitoring. PMID- 8412590 TI - [Evaluation of the level of endemic tuberculosis in a survey of Banqui (Central African Republic)]. AB - A cluster sample survey on tuberculosis has been carried out in Bangui in February 1988. The bimodal distribution of the diameters of IDR on children aged between 5 to 9, gives evidence of a circulation of some non typical mycobacteriae and enables to fix the limit of positivity at 14 mm. The prevalence rate of the tuberculotic infection is 7.9 +/- 1.7% in the surveyed children population at school. The annual risk of infection is evaluated at 1.09% that ranks the Centrafrican Republic in the countries with a low prevalence rate. PMID- 8412591 TI - [Unusual radiologic aspect of a hydatid cyst]. AB - The authors report us about an unusual hydatid cyst of the liver. Ultrasound, CT and MRI exams were used, and showed an unusual fatty component inside the lesion. The development of the disease in steatosic liver is not enough to explain the unusual density. PMID- 8412592 TI - [Utilization of laryngeal masks. Preliminary study. 21 cases. Department of Anesthesia-Resuscitation, Yaounde, Cameroon]. AB - The authors present a preliminary and retrospective study about the utilization of laryngeal mask. 21 patients underwent surgery concerning short or mid term intervention. A positioning of a laryngeal mask was easy with 83 p.c. of success. Although no shortcomings appeared in this series, one must know such a possibility and must take it into considerations. Not at all replacing endotracheal intubation, the laryngeal mask is in peculiar situations an other possibility of protection of respiratory tract, and of ventilation be spontaneous manual or mechanical. The advantages of laryngeal mask versus facial one are obvious. PMID- 8412593 TI - [Value of rectoscopy in the diagnosis of Schistosoma mansoni bilharziasis. Study at Nkolbisson-Cameroon]. AB - The aim of this study was to investigate endoscopically and histologically rectal mucosa complications in bilharzia patients. Coprology using KATO's technique was carried out in 1406 school children aged 4 to 18 years. 134 (9.53%) were infested. Among the infested school children and the sample population, a random selection of 80 school children (40 infested and 40 of sample population) was made, in whom rectoscopy and biopsy of rectal mucosa were carried out. The lesions found were classified on endoscopic and histologic basis. The prevalence of the disease 9.53% appears to be in constant regression. Inflammatory lesions are more common in bilharzia patients. Histology facilitated the unveiling of a specific rectitis in 72.5% bilharzia patients and 12.5% of the sample population. This study confirm the effectiveness of the several bilharzia eradication campaigns carried out in the Nkolbisson area. Rectoscopy with biopsy of the rectal mucosa should be reserved to symptomatic patients, in whom many coprologies have turned out negative. PMID- 8412594 TI - [Principal clinical manifestations during the course of disease caused by the human immunodeficiency virus (HIV) in Pointe-Noire (Republic of Congo). (307 cases hospitalized during 2 years at the medical service of the Regional Hospital of the Army)]. AB - At the sight of the hospitalization of 307 patients, attacked by AIDS, inside the department of Medicine of the Military Hospital in Pointe-Noire (Congo) between 1990-1992, the authors try to point out the principal epidemiological characteristics of their patients; give a semeiological and clinical descriptions of the key symptoms encountered; try to draw a scale depending on the apparition and frequency of the opportunistic infections. The great number of some diseases (tuberculosis, cryptococcal meningitidis, herpes zoster, diarrhea, neurologic complications ...) the more or less absence of others (pneumocystis carinii pneumonia) grant to Central Africa AIDS an undeniable originality compared to those of Europe and America. This analysis shows that a certain number of clinic signs in tropical area must attract the attention of the physician (facial palsy, herpes zoster, dementia, focal brain disorders), and so, to include the detection of AIDS, in the etiology of these affections. PMID- 8412595 TI - [Simple methods of chloroquine determination in urine and their adaptation in the field]. AB - Field malaria studies require often the on site detection of chloroquine in urine for monitoring chemoprophylaxis and treatment compliance, investigating drug-use patterns and screening prospective subjects for in vitro and in vivo chemosensitivity tests. The field adapted methods are described, analytical characteristics are compared and different approaches to field adapted assay of chloroquine are discussed. PMID- 8412596 TI - [Seroepidemiological survey of brucellosis in abattoir personnel in N'Djamena (Tchad)]. AB - The sample studied contained 214 people divided into 2 groups: 107 abattoir workers and 107 blood donors. All the sera were analyzed by testing with buffered antigen. The percentage of positive sera in the exposed population was 14% whereas it was zero in the group of blood donors. PMID- 8412597 TI - Correction of phase effects produced by eddy currents in solvent suppressed 1H CSI. AB - Accurate phasing of MRS spectra is often difficult unless time varying phase effects produced by gradient-induced eddy currents that persist during data acquisition are eliminated. This effect is particularly problematic in 1H-CSI spectra where frequency shifts produced by static field inhomogeneity and phase shifts produced by eddy currents combine. In this paper we present a method that corrects both shifts and eliminates manual phasing of individual CSI spectra typically required to recover a pure absorption line shape. The method uses a time domain phase correction derived from the ambient water signal acquired under identical conditions (i.e., acquisition parameters, gradient sequence) as the solvent-suppressed CSI data. Results from CSI experiments on phantoms and in vivo solvent suppressed 1H-CSI spectra from normal human brain are presented demonstrating the capabilities of the technique. PMID- 8412598 TI - Doxorubicin-mediated free radical generation in intact human tumor cells enhances nitroxide electron paramagnetic resonance absorption intensity decay. AB - The decay of nitroxide spin label electron paramagnetic resonance (EPR) absorption intensity was used to investigate the doxorubicin-mediated intracellular generation of free radicals. The effects of 50-500 micrograms/ml doxorubicin on human tumor cells (MCF-7, breast cancer cells, and HL-60, promyelocytic leukemia, cells) were studied by measuring 2,2,6,6 tetramethylpiperidine-1-oxyl (TEMPO) absorption intensity decay (TAID) at a TEMPO concentration of 10 microM. Doxorubicin accelerated the TAID in both cell lines with a detection limit of 50 micrograms/ml for MCF-7 cells and 500 micrograms/ml doxorubicin for HL-60 cells. Preincubation of cells with the iron chelating agent, deferoxamine (5 mM), partially prevented the effects of doxorubicin on the TAID. Catalase and copper, zinc-superoxide dismutase (Cu,Zn-SOD) had no influence on the effects of doxorubicin on the TAID in intact cells. However, Cu,Zn-SOD completely abolished the effects of doxorubicin on the TAID in a MCF-7 cell-free system. Our findings suggest that doxorubicin mediates the intracellular generation of O2.- and that iron is involved in this process. PMID- 8412599 TI - Spin-lattice relaxation time measurement by means of a TurboFLASH technique. AB - Rapid measurements of in vivo proton spin-lattice relaxation times (T1) in human tissues were performed by magnetic resonance imaging in a 1.5 T whole-body super conducting MR scanner. The measurements employ serial TurboFLASH imaging (Snapshot-FLASH) with scan times for a single experiment below 4 s. Using centric phase encoding order, an appropriate fitting of the T1-parameter from images with minimum motion artifacts is possible. Comparative T1-determination with a multipoint inversion recovery and spin-echo technique was performed on phantoms containing Gd-DTPA solutions with different T1-values. We found a maximum deviation of 3.3% for T1 < 1100 ms and of 12.1% for 1100 ms < T1 < 1700 ms from the results obtained by the IR technique. In vivo measurements of T1-relaxation times were performed in white and grey matter, cerebrospinal fluid, kidney, liver, spleen, muscle, and bone marrow, and yielded values that are in good agreement with reported data. PMID- 8412600 TI - Pulsatile flow artifacts in 3D magnetic resonance imaging. AB - Some of the important features of how pulsatile flow generates artifacts in three dimensional magnetic resonance imaging are analyzed and demonstrated. Time variations in the magnetic resonance signal during the heart cycle lead to more complex patterns of artifacts in 3D imaging than in 2D imaging. The appearance and location of these artifacts within the image volume are shown to be describable as displacements along a line in a plane parallel to that defined by the phase and volume encode directions. The angle of the line in the plane depends solely upon the imaging parameters while the ghost displacement along the line is proportional to the signal modulation frequency. Aliasing of these ghosts leads to a variety of artifact patterns which are sensitive to the pulsation period and repetition time of the pulse sequence. Numerical simulations of these effects were found to be in good agreement with experimental images of an elastic model of a human carotid artery under simulated physiological conditions and with images of two human subjects. PMID- 8412601 TI - Feature-recognizing MRI. AB - We describe a new theory of MR imaging that utilizes prior information in the form of a set of "training" images thought to be similar to the "unknown" objects to be scanned. First, the training images are processed to find an orthonormal series representation of these images that is more convergent than the usual Fourier series. The coefficients in this new series can be calculated from a subset of the phase-encoded signals needed to construct the Fourier image representation. The characteristics of the training images also determine exactly which phase-encoded signals should be measured in order to minimize error in the image reconstruction. The optimal phase-encodings are usually scattered nonuniformly in kappa-space. Good results were obtained when this theory was applied to imaging data from simulated objects and to experimental data from phantom scans. This theory provides the basis for developing efficient scanning and image reconstruction techniques that are "tailored" to each body part or to particular disease states. PMID- 8412602 TI - Apparent diffusion coefficient mapping of experimental focal cerebral ischemia using diffusion-weighted echo-planar imaging. AB - Diffusion-weighted, echo-planar imaging (EPI) was used to map regional changes in the apparent diffusion coefficient (ADC) during experimental focal ischemia in the rat brain following permanent middle cerebral arterial occlusion (MCAO). Sixteen 64 x 64 diffusion-weighted EPIs were acquired in 32 s with successively increasing amplitudes of the diffusion-sensitive gradient pulses. A linear least squares regression algorithm was used to fit 15 of the 16 two-dimensional matrices, on a pixel-by-pixel basis, to solve for the slope from which the ADC value was calculated. The correlation coefficient of the fit, R2, was used to filter the final ADC maps, and the ADCs were then scaled appropriately to be displayed in a 256 gray level format. Ranges (bins) of 0.05 x 10(-3) mm2/s were then grouped and color coded to qualify and quantify the evolution of ischemia in the MCA territory. The percentage of area in the ischemic and contralateral hemispheres in seven ADC bins were calculated at 30, 60, and 120 min after MCAO for 10 animals and demonstrated a significant increase in ADC bins below 0.45 x 10(-3) mm2/s and a decrease in bins above 0.50 x 10(-3) mm2/s over time. The postmortem infarct area, as measured by TTC staining, was highly correlated with the portion of the ischemic hemisphere falling below ADC values of 0.55 x 10(-3) mm2/s at 2 h after stroke onset. These studies suggest that focally ischemic brain tissue can be quantitatively subdivided according to ADC values and that ADC values below 0.55 x 10(-3) mm2/s 2 h following ischemia highly predict infarction in a rat permanent occlusion stroke model. PMID- 8412603 TI - Gadolinium enhancement in acute and chronic-progressive experimental allergic encephalomyelitis in the guinea pig. AB - Magnetic resonance imaging detects blood-brain barrier disruption after gadolinium-DTPA enhancement of central nervous system lesions of multiple sclerosis. Experimental allergic encephalomyelitis has many clinical and pathological features in common with multiple sclerosis including alterations in the integrity of the blood-brain barrier. We have compared T2-weighted cranial MR images with Gd-DTPA dimeglumine-enhanced T1-weighted images of myelin basic protein-induced acute (Days 8-42 postimmunization) and central nervous system induced chronic-progressive (Days 70-95 postimmunization) forms of experimental allergic encephalomyelitis in the guinea pig. Although animals from both groups had abnormal T2-weighted images, only the myelin basic protein-acute-EAE animals (Days 14-24 postimmunization) showed enhancement on postcontrast MR studies. The different responses of the acute and chronic diseases may result from different immunogens, severity of disease, or different permeability of the blood-brain barrier. Therefore, gadolinium-DTPA-enhanced, T1-weighted MR images distinguish acutely active and chronic inflammatory lesions in experimental allergic encephalomyelitis. PMID- 8412604 TI - Techniques for the measurement of the local myocardial extraction efficiency for inert diffusible contrast agents such as gadopentate dimeglumine. AB - We have previously shown that the concentration of Gd-DTPA as a function of time ([Gd-DTPA]t(t)) in the myocardium following an intravenous bolus injection of Gd DTPA can be modeled using the Modified Kety Equation. Fitting this model to measurements of [Gd-DTPA]t(t) in a region of myocardium would allow the determination of myocardial distribution volume (lambda) and the product of flow (F) and extraction efficiency (E), i.e., FE. Thus, to measure F, E must be known. We describe here techniques developed to measure local values of E in normal and diseased myocardium. These techniques are valid for any inert diffusible MR contrast agent. PMID- 8412605 TI - Measurement of the extraction efficiency and distribution volume for Gd-DTPA in normal and diseased canine myocardium. AB - We have previously shown that the myocardial Gd-DTPA concentration ([Gd DTPA]t(t)) after a bolus injection of Gd-DTPA can be predicted by the Modified Kety Equation (MKE). If [Gd-DTPA]t(t) can be determined by MRI and the data fit to the MKE, then the distribution volume (lambda) of Gd-DTPA and the myocardial flow (F) times the extraction efficiency (E), i.e., the FE product, can be determined. Therefore F can only be quantified if E is known. We measured the global E in vivo in normal canine myocardium, and measured E and lambda, in vitro, locally in normal, acute ischemic (n = 5; coronary artery occlusion < 4 h), infarcted (n = 4; coronary artery occlusion, 6 days) and reperfused (n = 4; coronary artery occlusion 2 h, and reperfusion 2 h and 6 days) myocardium. Results indicate that E differs with F and with individuals and consequently, F cannot be quantified using the MKE unless the local E is also determined in vivo. PMID- 8412606 TI - In vivo 31P NMR measurement of glucose-6-phosphate in the rat muscle after exercise. AB - Comparison of 31P NMR spectra of the rat gastrocnemius, obtained in vivo and from PCA extracts, after electrically induced contractions, demonstrates that glucose 6-phosphate (G6P) is the major metabolite in the low-field part of the PME spectral region. In vivo 31P NMR can thus be used to measure the muscle G6P concentration after exercise. PMID- 8412607 TI - Accurate measurement of T1 in vivo in less than 3 seconds using echo-planar imaging. AB - This paper describes a rapid and accurate method for measuring T1 in vivo using an echo-planar imaging version of the Look-Locher sequence. T1 values from 76 to 1330 ms have been measured in 3 s with a mean accuracy of 4.6%. PMID- 8412608 TI - Motion suppression improves quantification of rat liver volume in vivo by magnetic resonance imaging. AB - In response to the presence of certain compounds, rat liver weight can increase. Under the assumption that the liver density does not change, the liver volume will increase as well. To develop the capability to monitor this process noninvasively over time, we used liver volumes determined from MR images to estimate the in vivo liver volumes and weights of normal rats. We acquired multislice, spin-echo images from 18 rats using several protocols for suppression of motion artifacts. We found that volumes determined from data obtained using a combination of gradient moment nulling and respiratory gating, or a combination of signal averaging and "retarded" (after the pi pulse) phase-encoding, produced the most accurate estimates of in vivo liver volume and weight. PMID- 8412609 TI - Magnetic resonance imaging of perfusion in the isolated rat heart using spin inversion of arterial water. AB - Measurement of regional myocardial perfusion is important for the diagnosis and treatment of coronary artery disease. Currently used methods for the measurement of myocardial tissue perfusion are either invasive or not quantitative. Here, we demonstrate a technique for the measurement of myocardial perfusion using magnetic resonance imaging (MRI) with spin tagging of arterial water. In addition, it is shown that changes in perfusion can be quantitated by measuring changes in tissue T1. Perfusion images are obtained in Langendorff-perfused, isolated rat hearts for perfusion rates ranging from 5 to 22 ml/g/min. The MRI determined perfusion rates are in excellent agreement with perfusion rates determined from measurement of bulk perfusate flow (r = 0.98). The predicted linear dependence of the measured T1 (T1app) on perfusion is also demonstrated. The ability of perfusion imaging to measure regional variations in flow is demonstrated with hearts in which perfusion defects were created by ligation of a coronary artery. These results indicate that MRI of perfusion using spin inversion of arterial water gives quantitative maps of cardiac perfusion. PMID- 8412610 TI - Predicting RF field penetration in heterogeneous bodies using a 3-D finite element approach: preliminary results. AB - We present preliminary results for a 3D finite element calculation to evaluate RF penetration in conducting dielectric materials at high field strengths. A tetrahedral mesh is used along with a Coulomb gauge constraint in a finite element method that yields excellent numerical stability at high frequencies. Accuracy is verified by comparisons with analytic solutions for single-layer and multiple-layer heterogeneous systems and for a 3D spherical model. We have also compared the finite element model with experimental results presented by Foo et al., Magn. Reson. Med. 23, (1992). Agreement is very good and argues for the usefulness of the method in the calculation of RF penetration and RF power deposition effects in heterogeneous objects. PMID- 8412611 TI - Magnetic field distribution in models of trabecular bone. AB - A magnetostatic model consisting of a tetragonal lattice of struts of diamagnetic material, mimicking vertebral trabecular bone, was developed. The model allows estimation of the magnetic field histogram within the lattice's unit cell as a function of geometric parameters. The field was computed analytically from the induced magnetic surface charge density on the faces of the struts. The contribution from the induced magnetic field to the effective transverse relaxation rate, R2', was obtained as the mean decay rate of the Fourier transformed histograms, for both fixed and randomly oriented lattices. The model predicts the field distribution to increase with both strut thickness and density, paralleling material density. Finally, significant changes in R2' are predicted at constant material density, in that the field distribution widens with simultaneously increasing strut number density and decreasing strut thickness. PMID- 8412612 TI - Tesla gradient recalled echo characteristics of photic stimulation-induced signal changes in the human primary visual cortex. AB - Multi-echo measurements of photic stimulation-induced signal changes in human visual cortex were made at 4 Tesla in order to quantify the nature of the signal change and its vascular origin, and to determine the optimum echo time for detection of the changes. Utilizing high resolution images, two distinct regions (ascribed to be microvasculature and visible venous vessels) were identified as giving rise to the signal increase. The fractional signal changes in gray matter areas depended linearly on echo time (TE) in the range of 10 to 60 ms and extrapolated to virtually zero for TE = 0, indicating that in-flow effects secondary to stimulation-induced blood flow increases were negligible in our functional imaging studies; instead, signal change due to photic stimulation originated from the increase in the apparent transverse relaxation rate, 1/T2*. This decrease in (1/T2*), brought about by the alterations in hemodynamic parameters, was 1.3 +/- 0.4 s-1 for gray matter and 3.0 +/- 0.7 s-1 (averaged over 10 individuals) for venous vessels visible in the images. The optimum choice of echo time was found to be TE > or = T2*. PMID- 8412613 TI - Identification of vascular structures as a major source of signal contrast in high resolution 2D and 3D functional activation imaging of the motor cortex at 1.5T: preliminary results. AB - We have measured the T2* signal response associated with cortical activation due to finger motion at 1.5 Tesla. Both thin slice 2D and 3D images show signal intensity changes which vary from 2% to 32% depending on volunteer, echo time, slice thickness, and in-plane resolution. The largest signal change occurred for the thinnest slices and highest resolution (2 mm3). This is consistent with reducing partial volume effects and a simple difference in phase between the intravascular signal and surrounding parenchyma. No inflow enhancement was seen on the 2D or 3D scans, confirming the nature of the signal difference for this approach was due to local field inhomogeneity effects. Using 3D imaging, multiple effects can be seen simultaneously. With a 3D MRA method, it was possible to locate the vessel that was the source of the T2* behavior; it was in each case a vein on the surface of the cortical parenchyma. PMID- 8412614 TI - Localized real-time velocity spectra determination. AB - The accurate measurement of flow velocity has long been a subject of NMR research. In the field of medical imaging, a variety of techniques primarily based on the principle of Fourier encoding have been described. Due to time constraints, necessary trade-offs exist between spatial versus velocity spectral resolution. In general, either the average velocity of individual pixels is displayed or velocity spectral determinations are made at the cost of spatial localization. The recent development of multidimensional excitation pulses makes spatial localization possible during the excitation phase of the pulse sequences. This approach, coupled with time varying gradient readout, can be used to obtain single-shot localized velocity spectra. Using these concepts, we have obtained in vivo real-time measurements of localized velocity spectra on our clinical imager. PMID- 8412615 TI - Snapshot MRI with T2*-weighted magnetization preparation. AB - A method for encoding T2*-dependent contrasts into the preparation period of magnetization-prepared ultrafast MRI sequences is demonstrated and validated. The preparation block consists of a 90 degrees---tau---90 degrees p sequence, where p is shifted by 90 degrees for successive images. By adding two such successive images in quadrature, a combined image is obtained, in which the pixel intensities depend on the incoherent intravoxel dephasing that occurred during tau in a way that is similar to the dependence of TE in a gradient-echo sequence. The method could therefore be useful for applications that require very short repetition times for good temporal resolution, together with detection of T2* weighted contrasts, such as functional MRI in the brain. PMID- 8412616 TI - Evidence of Haemophilus ducreyi adherence to and cytotoxin destruction of human epithelial cells. AB - The adherence of ten different Haemophilus ducreyi strains to cultured human epithelial cells and the subsequent destruction of these cells was investigated in vitro using HEp-2 and HeLa cells. Bacterial adherence was measured with two assays, one employing viable bacteria and the other radiolabeled bacteria. In addition, the capacity of H. ducreyi to invade/penetrate the HEp-2 cells was examined. Differential interference contrast and transmission electron microscopy techniques were also used. In both cell lines, all ten strains of H. ducreyi manifested substantial adherence (the rates being 4-20% of the inoculum), irrespective of whether the bacteria were cultivated on solid or liquid media. Bacterial adherence reached a peak after about 2-3 h of incubation, though it was already manifest after only 15 min, a finding suggesting constitutive rather than inducible properties of H. ducreyi adhesins to be involved. The adherence capacity was diminished, but not totally abolished, when bacteria were heat treated at 100 degrees C for 30 min, indicating the adhesins to be fairly stable. On the other hand, treatment of HEp-2 cells with methanol, glutaraldehyde and emetine dichloride significantly reduced the adherence, indicating viable eukaryotic cells with native surface structures to be involved in bacterial adherence. This capacity of H. ducreyi to adhere to HEp-2 cells was confirmed both by electron microscopy and by differential interference microscopy. Some adherent bacteria were also capable of penetrating epithelial cells, as observed with an invasion assay and confirmed by transmission electron microscopy. Further incubation of the cell monolayers with the ten strains resulted in the cell-death and total damage of monolayers for seven cytotoxin-producing strains, indicating cytotoxin action to be responsible for the destruction of the monolayer. All strains manifested capacity to survive and multiply on the cell monolayer. We propose the first step in the pathogenesis of chancroid to be the adherence of bacteria to epithelial cells, followed by the action of cytotoxin and further bacterial proliferation. This sequence of events is suggested to result in the production of genital ulcers by H. ducreyi organisms. PMID- 8412617 TI - Role of Shigella dysenteriae type 1 slime polysaccharide in resistance to serum killing and phagocytosis. AB - Shigella dysenteriae type 1 produce a slime polysaccharide when cultivated in vivo in adult rabbit ileal loops or in vitro in casamino acid yeast extract broth medium which promotes hemagglutination of these bacteria. Seven strains of S. dysenteriae 1 grown in vitro and in vivo and possessing slime polysaccharides resisted killing by normal human serum as compared to bacteria grown under conditions which do not stimulate the production of capsular polysaccharide and did not resist serum killing (mean survival 72% for in vitro growth and 73% for in vivo growth conditions favoring capsule production vs < 2% for growth conditions which do not favor capsule production; P < 0.001 for both comparisons). Similar differences were observed when killing was assessed by phagocytosis (62-78% vs < 2%; P < 0.001). We conclude that capsular polysaccharide may be an additional virulence factor of S. dysenteriae 1. PMID- 8412618 TI - Genetic analysis of the diversity in outer membrane protein P2 of non encapsulated Haemophilus influenzae. AB - The molecular basis for the diversity of the major outer membrane protein (MOMP) P2 of non-encapsulated Haemophilus influenzae was analyzed by direct sequencing of fragments of the P2 genes, obtained by the polymerase chain reaction. Genetically divergent H. influenzae strains were isolated from patients with otitis media and from infected patients with chronic obstructive pulmonary disease (COPD). The nucleotide sequences of these P2 genes were determined and compared with P2 gene sequences of three non-encapsulated strains and with the P2 gene of encapsulated H. influenzae type b. Variation in the nucleotide sequence of the P2 genes of non-encapsulated H. influenzae was localized in eight distinct regions which were interchanged with relatively conserved regions. Regions 2, 4, 5 and 8 were found to be hypervariable, since they were characterized by multiple deletions or substitutions of nucleotides. According to the proposed structure of MOMP P2 the variable regions were localized in eight loop structures exposed at the outside of the outer membrane. Loops 2, 4, 5 and 8 were hypervariable in all isolates whereas loop 6 varied only in isolates from COPD patients with persistent infections. PMID- 8412619 TI - Effect of late administration of anti-TNF alpha antibodies on a Salmonella infection in the mouse model. AB - The effect of late administration of anti-TNF alpha antibodies on the course of a Salmonella infection in mice was evaluated. Administration of anti-TNF alpha antiserum as late as day 5 after challenge enhanced a sublethal primary infection with the virulent Salmonella typhimurium C5 in innately resistant (Ityr) A/J mice by preventing the suppression of exponential bacterial growth in the reticuloendothelial system (RES) (plateau phase). When the anti-TNF alpha treatment was started well after the establishment of the plateau (day 7) a prompt relapse of the infection occurred, with the rapid resurgence of bacterial growth in the reticuloendothelial system leading to the death of the animals. In contrast, late administration of the antiserum did not affect the clearance from the tissues of an avirulent temperature-sensitive mutant of S. typhimurium C5 (C5TS). Innately susceptible (Itys) BALB/c mice immunized with the SL3261 aroA live vaccine acquire solid long-lasting protection from oral challenge with the virulent C5 strain, suppressing growth of the challenge in the RES. Administration of anti-TNF alpha antibodies on day 8 of a secondary oral infection with strain C5 abrogated vaccine-induced protection, with a progressive increase of bacterial numbers in the RES leading to the death of the animals. The results indicate that TNF alpha is constantly required for the control of virulent salmonellae in the RES, both in a sublethal primary infection in innately resistant mice and also in a secondary infection in innately susceptible mice immunized with a live vaccine. TNF alpha may not be essential for bacterial clearance of avirulent organisms from the tissues. PMID- 8412620 TI - Site of transcriptional activation of virB on the large plasmid of Shigella flexneri 2a by VirF, a member of the AraC family of transcriptional activators. AB - VirB plays a central role in the regulation of virulence of Shigella flexneri. It acts as a transcriptional activator and is itself transcriptionally activated by another virulence protein, VirF. Experiments were performed in order to identify the site upstream of virB at which VirF binds in order to activate transcription. Progressive 5' deletions of the DNA upstream of the transcription start point of virB were constructed by subcloning and Bal31 deletion. These deletion derivatives were cloned into the chloramphenicol acetyltransferase (CAT) reporter gene plasmid pKK232-8 and the resulting plasmids were analysed using a CAT activity assay. This allowed identification of minimal regions required for VirB promoter activity and regions required for full enhancement of promoter activity by VirF. A region approximately 100 bp upstream from the transcription start point of virB was identified as being necessary for full activation of this promoter by VirF. This region encompasses at least one inverted repeat which may play a role in transcription repression in the absence of the activator protein, VirF. PMID- 8412621 TI - Virulence of a Bordetella pertussis strain expressing a mutant adenylyl cyclase with decreased calmodulin affinity. AB - Bordetella pertussis, the pathogen responsible for whooping cough, produces a toxic calmodulin-sensitive adenylyl cyclase which enters animal cells and increases intracellular cAMP. A point mutant of B. pertussis with abolished adenylyl cyclase catalytic activity was over 1000-fold less pathogenic to newborn mice than wild-type bacteria, demonstrating the importance of the adenylyl cyclase for B. pertussis virulence (Gross et al.). The B. pertussis adenylyl cyclase is highly sensitive to calmodulin with an apparent Kd for calmodulin of approximately 1 nM. The importance of this high-affinity calmodulin binding for virulence in vivo was examined by the creation of a B. pertussis point mutant (Trp-242 to Glu-242) with 200-fold lower calmodulin affinity than the native enzyme. This mutant B. pertussis strain retained its virulence in a newborn mouse model of pertussis, but the time course for establishment of a lethal infection in vivo was significantly delayed for the mutant strain. These data illustrate that high-affinity calmodulin binding is not obligatory for the activity of this toxin but is important for the rate for establishment of a lethal infection. PMID- 8412622 TI - IFN-gamma produced in vivo during the first two days is critical for resolution of murine Leishmania major infections. AB - Resolution of murine infection with Leishmania major is dependent upon the presence of IFN-gamma during the first week of infection. To more precisely determine the period during which IFN-gamma is critical, we infected footpads of resistant C3H/HeN mice with amastigotes of L. major and treated these mice with neutralizing monoclonal Ab (MAb) specific for IFN-gamma on successive days. Mice treated with anti-IFN-gamma MAb on or before day 2 had significantly enlarged lesions, and increased parasites in lesions, compared with mice treated with an isotype control MAb. In contrast, mice treated with anti-IFN-gamma MAb on day 3 or later resolved their lesions and had no parasites at the inoculation site. Related experiments obtained with a neutralizing MAb specific for TNF-alpha demonstrated the critical role TNF-alpha plays in resolution of Leishmania infection. Thus, IFN-gamma and TNF-alpha were both critical for resolution of infection with L. major, with IFN-gamma's role limited to the first 2 days. PMID- 8412623 TI - Increased expression of heat shock protein, transferrin, and beta 2-microglobulin in astrocytes during scrapie. AB - Scrapie is a slow infection and neurodegenerative disease of animals characterized pathologically by formation of amyloid, astrocytosis, vacuolation and injury and death of neurons. Our previous studies of scrapie point to: (i) a critical role for the astrocyte in responding to, and perhaps inadvertently contributing to, the neuropathological manifestations of infection; and (ii), the hypothesis that the astrocyte executes a programed response to neurological injury analogous to the stress response. The expression of genes encoding transferrin and beta 2-microglobulin has been shown to increase during scrapie and we sought in the studies reported here to determine whether this modulated expression would map to astrocytes. We also looked for changes in a heat shock protein that is induced in the stress response. We show that transferrin, beta 2 microglobulin and heat shock 72 kD protein all increase in astrocytes in the course of infection. We speculate in the discussion on the possible functions of these and other proteins in neurodegenerative processes and why these functions so frequently reside in the astrocyte. PMID- 8412624 TI - Detection of murine cytomegalovirus (MCMV) DNA in skin using the polymerase chain reaction (PCR). AB - We used the polymerase chain reaction (PCR) and primers for the immediate early gene of murine cytomegalovirus (MCMV) to detect MCMV DNA in skin harvested from mice during acute infection. MCMV DNA was also detected in DNA extracted from spleen and salivary gland of MCMV-infected mice, but not in the skin, salivary gland, or spleen of uninfected, seronegative mice. Detection of MCMV DNA in skin provides direct evidence that skin can serve as a vehicle for transmission of MCMV. This observation is relevant to humans, such as burn patients, who receive skin allografts that may be infected with cytomegalovirus. PMID- 8412625 TI - Role of plasmid-encoded antigens of Yersinia enterocolitica in humoral immunity against secondary Y. enterocolitica infection in mice. AB - In the present study the role of Yersinia enterocolitica antigens in humoral immunity against secondary Y. enterocolitica infection has been investigated. For this purpose, BALB/c mice, 4 weeks after immunization by primary infection with a sublethal dose of various Y. enterocolitica strains and mutant strains, were challenged with a lethal dose of highly virulent Y. enterocolitica strains of serotype O:8. As evident from the survival rate and protection against bacterial growth in the spleens of mice, only immunization with wild-type or attenuated strains harbouring an intact virulence plasmid mediated protection against a lethal challenge. Protection by immunization with plasmid-harbouring strains correlated with persistence of the bacteria in spleens for at least 7 days after immunization and high serum titres of Yersinia-specific antibodies directed against both chromosomal and plasmid-encoded determinants. Furthermore, the adoptive transfer of the purified IgG fraction of a rabbit serum specific for the adhesin YadA encoded by virulence plasmid pO8 from Y. enterocolitica O:8 mediated protection against a lethal challenge with the serotype O:8 strain harbouring the virulence plasmid pO8 but did not mediate protection when this strain harboured the virulence plasmid pO9 from serotype O:9. In summary, the results demonstrate that in our experimental mouse infection model: (i) the presence of the intact virulence plasmid in an immunizing Y. enterocolitica strain is essential for induction of protection against a lethal challenge with highly virulent Y. enterocolitica and (ii) that antibodies against plasmid-encoded determinants of Y. enterocolitica, especially YadA, are of major importance for achievement of protective immunity in mice. PMID- 8412626 TI - Cloning of two distinct hemolysin genes from Porphyromonas (Bacteroides) gingivalis in Escherichia coli. AB - Hemolysin is considered a potent virulence factor in a large number of Gram positive and Gram-negative bacterial pathogens. The hemolysin produced by the oral pathogen Porphyromonas gingivalis functions to provide the cell with its required heme-containing molecules for growth in the periodontal pocket. Two distinct P. gingivalis genes, each of which confers a hemolytic phenotype in Escherichia coli, were isolated by screening genomic DNA libraries of P. gingivalis on sheep blood agar plates. The results obtained from physical maps and Southern blots indicated a considerable degree of divergence in the nucleotide sequences of these two genes. Maxicell analyses of the recombinant plasmids in E. coli suggested that plasmid pPGH5 encoded a polypeptide of molecular weight 48 kDa, while an 18-kDa polypeptide was obtained with pPGH1 and pPGH7. When E. coli harboring these hemolysin genes were subjected to iron starvation, the levels of hemolysin activity increased. Biochemical characterization of hemolytic activities indicated that the activity of both hemolysins was inhibited by Mg2+ and Ca2+; but not by EDTA. Elevated levels of hemolytic activity were obtained from the E. coli recombinant strains in the presence of glutathione, DTT and 2-mercaptoethanol. Cholesterol inhibited the activity. PMID- 8412627 TI - Neisseria gonorrhoeae strain MS11 harbouring a mutation in gene aroA is attenuated and immunogenic. AB - An aroA mutant of gonococcal strain MS11 was constructed (JKD298) and compared with the wild-type (JKD288). The requirement of JKD298 for aromatic compounds, typical of an aroA mutant, was demonstrated using defined media. Other than the expected auxotrophy, no further differences could be demonstrated between the parent and the aroA mutant. SDS-PAGE analysis of protein and lipopolysaccharide (LPS) profiles showed no differences between the strains. Bactericidal assays using human and guinea-pig normal sera showed that both strains were serum sensitive and were similarly converted to serum resistance by in vitro sialylation using CMP-NANA. Infectivity experiments in guinea-pig subcutaneous chambers showed considerably reduced virulence of the aroA JKD298, which could only infect chambers at very high doses. Established infections by either strain elicited a strong PMN response in which similar proportions of each strain were seen intracellularly. Infections by JKD298 provoked a strong antibody response as detected by ELISA using whole sonicated gonococci. This is the first demonstration of attenuation of Neisseria gonorrhoeae by introduction of a defined mutation in a metabolic gene. PMID- 8412628 TI - Molecular cloning and expression of Rickettsia prowazekii genes for three outer membrane proteins in Escherichia coli. AB - Rickettsia prowazekii (virulent Breinl strain) random genomic DNA fragments were cloned in the lambda gt11 expression vector by using non-palindromic adaptors. Several immunoreactive clones were selected after screening 20,000 individual recombinant plaques with human convalescent serum. Some recombinants expressed complete 60 kDa polypeptide, and others expressed beta-galactosidase fusion polypeptides containing different epitopes of 134 kDa protein of the R. prowazekii outer membrane (OM). Amplified genomic library was screened with monospecific antibodies directed against abundant 31 kDa and 29.5 kDa OM proteins (OMPs). Several recombinant clones expressing full or part of 29.5 kDa polypeptide, and none expressing 31 kDa polypeptide were revealed. The serum of a patient convalescing from epidemic typhus did not react in Western blot with recombinant 29.5 kDa protein. PMID- 8412629 TI - Polymerase chain reaction amplification, cloning and sequencing of variant Escherichia coli Shiga-like toxin type II operons. AB - The polymerase chain reaction (PCR) was used to amplify approximately 1.5 kb segments of DNA containing complete Shiga-like toxin type II operons from Escherichia coli serotypes OX3:H21 and O111:H-. These fragments were cloned and DNA sequence analysis identified further variations compared with published SLT II sequences. PMID- 8412630 TI - Forty-two years of peripheral nerve surgery. PMID- 8412631 TI - Microvascular surgery in Europe: the early achievements and development. PMID- 8412632 TI - Prefabricated microsurgical tissue units: progress over the past 9 years. PMID- 8412633 TI - Percentage of nerve injuries in which primary repair can be achieved by end-to end approximation: review of 2,181 nerve lesions. AB - Primary nerve repair yields better results than secondary reconstruction but is not always possible. We reviewed a series of 2,181 fresh nerve injuries of the upper limb. One nerve only was injured in 41% of the patients; two or more in 59%. One thousand four hundred eighty-two injuries (68%) were located in the digits. The injured limb segment was lost or beyond repair in 387 cases (18% of all cases). In the 1,794 remaining injuries, primary treatment was accomplished by end-to-end suture 1,568 times (87%) and by graft 33 times (> 2%) and was impossible in 193 cases (11%). PMID- 8412634 TI - Bridging nerve defects with combined skeletal muscle and vein conduits. AB - The use of vein or muscle grafts to bridge nerve defects longer than 1-1.5 cm gives poor results. Veins collapse and in muscle grafts axons may regrow outside the graft. We used veins (to guide regeneration) filled with muscle (to avoid vein collapse). Nerve regeneration through 1 and 2 cm grafts made of vein plus muscle was compared with similarly long traditional nerve grafts, free fresh muscle grafts, and empty vein grafts. Regeneration was assessed clinically and histologically (qualitative and quantitative evaluation) in the graft and distal nerve stumps. Vein plus muscle grafts were superior to vein and fresh muscle grafts both functionally and histologically. Functional results were similar to those found in traditional nerve grafts, but axon number was superior in the veins filled with muscle. This suggests that vein filled with muscle might serve as a grafting conduit for the repair of peripheral nerve injuries and could give better results than traditional nerve grafting. PMID- 8412635 TI - Congenital pseudarthrosis of the forearm: treatment of six cases with vascularized fibular graft and a review of the literature. AB - Six cases of pseudarthrosis of the forearm are reported. One involved only the radius, one the ulna alone, and four both forearm bones. There was associated neurofibromatosis in five cases. All the cases were treated by wide resection and free vascularized fibular graft. In two cases, a single-bone forearm was created. In all cases, good results were achieved, with moderate forearm shortening. The literature suggests electrical stimulation, corticoperiosteal graft, osteotomy of the involved bone, and free vascularized bone grafting as treatment alternatives. Vascularized free bone grafting appears to be more successful than other treatments. The use of microsurgical sutures permits this treatment in young patients. PMID- 8412636 TI - Wound closure at the trunk by microvascular free flap transfer. AB - Defects of the thoracic or abdominal wall can be congenital or caused by trauma or tumour resection. There may be other problems, such as infection and irradiation effects. In most cases those defects can be closed by local cutaneous and fasciocutaneous or by muscle and myocutaneous flaps. In some rare instances, the use of pedicled flaps may be limited. The size of the defect, the impossibility of closing the donor site, an impaired blood supply, or poor quality of local tissues represent such limitations. In these cases microvascular flap transfer has enlarged our choice of alternative methods. Advantages of this method are the one-stage procedure and the wound coverage by well-vascularized tissue. This is especially beneficial in areas of infected or irradiated tissue. On the other hand microvascular flap transfer requires high technical skill and extensive perioperative and postoperative care. PMID- 8412637 TI - Total scalp replantation based on one artery and one vein. AB - A successful replantation of a totally avulsed scalp, including both eyebrows, with only one arterial and one venous anastomosis to the superficial temporal vessels is described. Apart from a small partial skin necrosis of the right eyebrow, the entire transplant survived. Subsequently, the patient required only minor additional skin grafting, correction of scars with a skin expander, and reconstruction of the eyebrow with hair micrografts. Replantation of the total scalp based on two or more vessels has previously been recommended and reported elsewhere. In the present case, complete survival of the scalp on only one artery and one vein was demonstrated, indicating that replantation should be considered even if available vessels for anastomosis are minimal. PMID- 8412638 TI - Histological evaluation of free revascularized iliac crest bone after transplantation for mandibular reconstruction. AB - Differing views on the nutritional status of revascularized bone in animal studies and in human transfer have been expressed. Normal viability is reported as well as single cell necrosis and devitalized areas and, finally, necrosis of the complete graft. From 19 patients with revascularized bone transfer into injured recipient tissues, we obtained 32 drill biopsies out of the centre of the graft and near the mandibular stump during the removal of the osteosynthesis plates. The histological preparations were examined for bone viability. In 58% of all patients, there occurred partial necroses in the grafted bone and in 26% single cell necroses; 16% of the transplants maintained normal viability. The areas with disturbed nutrition were found more frequently in peripheral areas than in central regions of the graft (p < 0.05, chi 2 test). The overall graft failure rate was 6.8%. The results of this clinical study are discussed with particular reference to the technical procedure, the recipient tissue quality, and the design of the graft. The indication for using revascularized bone transfer in difficult local conditions of the recipient tissues has been confirmed. The viability of the grafts cannot always be maintained entirely. The particular risk to the viability of the graft in the peripheral regions requires careful technical procedures. PMID- 8412639 TI - Microsurgery in gaining paediatric vascular access for haemodialysis. AB - One hundred and one surgical procedures performed in children for construction and maintenance of vascular accesses for haemodialysis were retrospectively analyzed. There were 86 operations performed to create a new fistula in patients without vascular access or with nonrecoverable failed angio-access. Fifteen surgical procedures were performed to treat fistula complications. The new fistulas were radiocephalic n = 60 (70%), ulnar-basilic n = 5 (5.8%), antecubital n = 9 (10.3%), and PTFE grafts n = 12 (14%). Microsurgical techniques were used in all cases, including PTFE graft fistulas. A microscope was used in 56 cases (55.4%) and magnifying loupes (x 2.5 magnification) in the rest of the operations. Early-failure rate for radiocephalic fistulas was 10%. Cumulative patency rates in radiocephalic fistulas were 79%, 75%, and 70% at 1, 2, and 5 years, respectively. No statistical differences were found from the cumulative patency curve of 730 radial-cephalic fistulas performed in adults during the same period of time. Radiocephalic fistulas can be constructed in most paediatric cases using microsurgical technique. Elbow fistulas can be the second-choice vascular access, and PTFE grafts can be reserved for children with exhaustion of autologous veins. Brachial-jugular PTFE grafts can be used in cases of subclavian vein stenosis. PMID- 8412640 TI - Aneurysmal cyst of the proximal radius: resection and free vascularized fibular bone graft. AB - We present the case of a 22-year-old woman with an aneurysmal cyst of the right proximal radius, treated by resection and a free bone graft (microvascular fibular transfer), without recurrence after 4 years. Over a 4 month period, there was a rapidly expanding and lytic lesion found in the radius. There were symptoms of elbow and wrist pain and early radially innervated muscle weakness. Radiological examination showed a localized tumour of the proximal radius, type 2, grade C, according to the classification of Campanacci. Biopsy led to the diagnosis of an aneurysmal bone cyst. Complete tumour resection was followed by a free vascularized fibular bone graft, resulting in good functional improvement, without local recurrence. Aneurysmal bone cysts are a rare condition, and the location in the radius has not previously been described. Vascularized bone grafts are mechanically and biologically superior in this type of case. PMID- 8412641 TI - Neurotizations by means of the cervical plexus in over 100 patients with from one to five root avulsions of the brachial plexus. AB - In the years 1982-1991, we treated over 100 patients with avulsion of from one to five roots in perinatal (n = 21) and traumatic (n = 81) lesions of the brachial plexus. Based on the clinical diagnosis and the surgical findings, we divided all operated patients into four groups: 1) avulsion of all roots or avulsions and disruption of the rest of the brachial plexus; 2) avulsion and disruption of one to four roots and preservation of some elements of the brachial plexus with fibrotic changes; 3) avulsion and disruption of one to three roots, with some elements showing no macroscopic changes; and 4) children with perinatal brachial plexus palsy. In cases of root avulsion, we performed neurotization from the cervical plexus, the accessory and intercostal nerves. In patients with one to three root avulsions, we have performed selective neurotization for 3 years. The best results were obtained in groups 3 and 4, but some improvement was also noted in the other groups. PMID- 8412642 TI - Toxic oil syndrome. A long-term follow-up of a cohort of 332 patients. AB - Toxic oil syndrome is a multisystemic, epidemic disease that appeared in Spain in 1981, related to the intake of rapeseed cooking oil sold in bulk. It affected 19,748 people, of whom 457 died. The toxic substance was never identified. We report the 8-year follow-up of a cohort of 332 patients. The disease was usually severe and disabling during the first 2 years, but the clinical condition of most of the patients improved thereafter. The acute phase lasted 2 months, and was characterized by pulmonary edema, rash, eosinophilia, and myalgia. During the intermediate phase (second to fourth months), severe myalgia, skin tenderness, subcutaneous edema, altered liver function, and pulmonary hypertension developed. Later on, an early chronic phase developed, from the fourth month to the end of the second year. It was marked by scleroderma, sicca syndrome, polyneuropathy, joint contractures, weight loss, and functional limitations. The clinical manifestations improved during the late chronic phase. Its most prominent clinical features were muscle cramps, chronic musculoskeletal pain, chronic lung disease, Raynaud phenomenon, carpal tunnel syndrome, and psychologic disturbances. Only 9% of the patients achieved remission after the acute phase, the rest developing late clinical manifestations of the disease. The severity of the chronic manifestations was rather variable. At the end of the 8-year follow up, there were 10 TOS-related deaths (3%), 47% of the patients had some kind of complaint, albeit subtle in most cases, and 16% showed organic involvement related to TOS. The most important pathologic features of TOS were widespread interstitial infiltrates, non-necrotizing angiitis, endothelial proliferation, and tissue fibrosis. Toxic oil syndrome is a dramatic example of an induced scleroderma-like syndrome, similar to the eosinophilia-myalgia syndrome. Patients with EMS may develop some of the late clinical features of TOS in the years to come. PMID- 8412643 TI - Polyarticular septic arthritis. AB - Twenty-five cases of polyarticular septic arthritis (PASA) were observed in our department over a 13-year period. They accounted for 16.6% of all septic arthritis (15% on average in the literature). A male predominance was noted in our patients, as well as in the literature. The knee was the most frequent location followed by the elbow, shoulder, and hip, in varying order depending on the series. An average of 4 joints was involved. The causative microorganism was Staphylococcus aureus in 20/25 of our patients and in about 50% of published cases. Other frequently causative organisms were streptococci and gram-negative bacteria. Blood cultures and joint aspirations were positive in 19/22 and 23/25 of our cases, respectively. Other septic lesions were noted in 10/25 of our cases. Fever and severe leukocytosis were absent at admission in 5/25 (literature, 37%) and 10/25 of our 25 patients, respectively. The underlying disease was rheumatoid arthritis in 13/25, while 9 of the other patients had immunodepression caused by drugs or by concurrent illness. Typically, rheumatoid arthritis was long-standing and erosive, patients having ulcerated calluses on the feet. This skin source was also noted in 23/36 published cases of PASA in rheumatoid arthritis. Systemic lupus erythematosus was an uncommon disease in PASA, but its presence promoted gram-negative infection. Despite effective therapy with 2 antibiotics, 8/25 patients died, a prognosis that is equally severe in cases reported in the literature (30%) and one that has remained surprisingly stable over the last 40 years. For comparison, the death rate was only 4% in our patients with MASA. Factors contributing to a poor prognosis were age greater than 50 years, rheumatoid arthritis as an underlying disease, and disease of staphylococcal origin. Septic polyarthritis should be considered even when the clinical picture is not florid--when patients have low fever and normal white blood cell counts. Nor should the simultaneous involvement of distant joints rule out infection. Indeed, the frequency of underlying rheumatic disease and its treatment may further confuse the clinical presentation. Joints suspected of harboring infection should be aspirated, including those previously affected by the concurrent rheumatism. PMID- 8412644 TI - Blastomycosis in immunocompromised patients. AB - Among the endemic mycoses, blastomycosis has been least often associated with disorders of immune function, but the data presented herein suggest that blastomycosis may occur more commonly in immunocompromised patients than was previously recognized. We have observed a marked increased in the number of immunocompromised patients with blastomycosis over the last 15 years, increasing from about 3% of patients seen between 1956 and 1977 to almost 24% patients seen between 1978 and 1991. The disease appears to be much more aggressive in immunocompromised than in normal hosts. Almost 30% of the patients in our series died secondary to blastomycosis, with most deaths occurring within 5 weeks following the diagnosis. Furthermore, almost one third of those patients who died of other causes had evidence of persistent blastomycosis at death. Multiple organ and central nervous system involvement were relatively common in this series. For these reasons, early and aggressive therapy with amphotericin B is indicated for most immunocompromised patients with blastomycosis. Oral therapy with an azole compound should probably be reserved for patients who have responded to a primary course of amphotericin B but who require additional or long-term suppressive therapy. Until more data are available, the newer azoles should be used with caution as primary therapy in immunocompromised patients with blastomycosis, and considered only in patients with limited disease and a stable underlying condition. Caring for the immunocompromised patient poses many diagnostic and therapeutic challenges to the clinician, and among those patients who have been exposed to areas endemic for blastomycosis, B. dermatitidis must be regarded as a potentially important opportunistic pathogen. PMID- 8412645 TI - Spondyloepimetaphyseal dysplasia: clinical and radiologic investigation of a large kindred manifesting autosomal dominant inheritance, and a review of the literature. AB - Spondyloepimetaphyseal dysplasias (SEMDs) are a heterogeneous group of skeletal disorders that can have a genetic basis, but their classification and prognostication suffer because few families have been extensively studied. We describe a large kindred affected by a unique type of SEMD that is transmitted as an autosomal dominant trait. The propositus and his affected brother and first cousin were evaluated as inpatients. Other kindred members were screened by telephone interviews and lateral thoraco-lumbar spine radiographs, and then, in most cases, investigated by additional x-ray studies. Of the 29 living members of the kindred, 22 were studied radiologically. Among the 22 subjects investigated, 15 were affected, and the status of 1 individual with minor changes on x-ray was indeterminate. The deceased patriarch was presumed to be affected. These 16 affected subjects could usually, but not invariably, be distinguished from their unaffected sex-matched siblings by their smaller heights. Nevertheless, it was only affected children who had short stature; the heights of all affected adults were normal. Often, affected individuals had rhizomelic shortening, especially of the lower extremities, and genu varum (not always evident clinically, but present on radiographs). Occasionally, they also manifested limited extension of their upper limbs. Radiologic study showed abnormal metaphyses, epiphyses, and vertebrae in affected children, but these 3 skeletal regions became less remarkable by late childhood and most affected adults had normal epiphyses. One obligate affected man had only spinal changes. Despite their normal heights, severely affected adults who had bowing deformity of their legs developed disabling degenerative joint disease limited to the knees in the 7th decade of life--disease severe enough to require knee replacement surgery. PMID- 8412646 TI - Molecular drift of the bride of sevenless (boss) gene in Drosophila. AB - DNA sequences were determined for three to five alleles of the bride-of-sevenless (boss) gene in each of four species of Drosophila. The product of boss is a transmembrane receptor for a ligand coded by the sevenless gene that triggers differentiation of the R7 photoreceptor cell in the compound eye. Population parameters affecting the rate and pattern of molecular evolution of boss were estimated from the multinomial configurations of nucleotide polymorphisms of synonymous codons. The time of divergence between D. melanogaster and D. simulans was estimated as approximately 1 Myr, that between D. teissieri and D. yakuba as approximately 0.75 Myr, and that between the two pairs of sibling species as approximately 2 Myr. (The boss genes themselves have estimated divergence times approximately 50% greater than the species divergence times.) The effective size of the species was estimated as approximately 5 x 10(6), and the average mutation rate was estimated as 1-2 x 10(-9)/nucleotide/generation. The ratio of amino acid polymorphisms within species to fixed differences between species suggests that approximately 25% of all possible single-step amino acid replacements in the boss gene product may be selectively neutral or nearly neutral. The data also imply that random genetic drift has been responsible for virtually all of the observed differences in the portion of the boss gene analyzed among the four species. PMID- 8412647 TI - The Drosophila ribosomal protein S6 gene includes a 3' triplication that arose by unequal crossing-over. AB - Ribosomal protein S6 (rpS6) is the major phosphoprotein of the small ribosomal subunit of eukaryotes and is phosphorylated in response to treatment with mitogens and other stimuli. We have examined the organization of the rpS6 gene of Drosophila melanogaster. Comparisons of a cDNA with genomic DNA identify a transcription unit including three exons. Two tandem repeats downstream of this transcription unit reiterate divergent copies of the third exon and flanking regions. Comparisons of these three repeats with respect to nucleotide base substitutions and deletions or insertions show clearly that they arose via a duplication and subsequent crossing-over between misaligned copies. Although no direct evidence exists that the downstream exons are transcribed, the maintenance of open reading frames in spite of extensive genetic changes is consistent with a protein-coding function. PMID- 8412648 TI - Positive selection for colicin diversity in bacteria. AB - To examine the hypothesis that colicin proteins are subject to diversity enhancing selection, we studied the rates of synonymous, nonsynonymous, and intergenic nucleotide substitution in three pairs of closely related colicin clusters. The results indicate that the immunity gene and the immunity-binding domain of the colicin gene, which interact to provide specific immunity from the lethal action of the colicin toxin, accumulate substitutions at synonymous and nonsynonymous sites several times more rapidly than does the remainder of the colicin cluster. We suggest that this increased level of divergence, centered at the immunity protein, may be the result of the combined action of recombination and positive selection acting to increase colicin diversity in natural populations of Escherichia coli. PMID- 8412649 TI - Major-histocompatibility-complex variation in two species of cichlid fishes from Lake Malawi. AB - Lake Malawi in eastern Africa harbors > 500 endemic species of cichlid fishes, all of which are believed to have emerged from a single founding population in the past 2 Myr. Molecular characterization of differences among the species could provide important information about the nature of speciation in the period of adaptive radiation. Because of the close relationship, however, molecular variation among the species has been difficult to ascertain. In this communication, we provide evidence for extensive differences, in major histocompatibility-complex (Mhc) class II genes, between two related species, Pseudotropheus zebra and Melanochromis auratus. We used specific primers to amplify and sequence intron 1 and exon 2 of the class II genes from 18 individuals. Although we found 20 different sequences among the 42 that we produced, there was not a single sequence shared by the two species. Thus the study suggests that different cichlid species of Lake Malawi have different profiles of class II alleles, presumably because the polymorphism present in the ancestral founding population segregated differentially into the various species. These results make Mhc genes an important tool for elucidating speciation. PMID- 8412650 TI - Theoretical foundation of the minimum-evolution method of phylogenetic inference. AB - The minimum-evolution (ME) method of phylogenetic inference is based on the assumption that the tree with the smallest sum of branch length estimates is most likely to be the true one. In the past this assumption has been used without mathematical proof. Here we present the theoretical basis of this method by showing that the expectation of the sum of branch length estimates for the true tree is smallest among all possible trees, provided that the evolutionary distances used are statistically unbiased and that the branch lengths are estimated by the ordinary least-squares method. We also present simple mathematical formulas for computing branch length estimates and their standard errors for any unrooted bifurcating tree, with the least-squares approach. As a numerical example, we have analyzed mtDNA sequence data obtained by Vigilant et al. and have found the ME tree for 95 human and 1 chimpanzee (outgroup) sequences. The tree was somewhat different from the neighbor-joining tree constructed by Tamura and Nei, but there was no statistically significant difference between them. PMID- 8412651 TI - Prospects for estimating nucleotide divergence with RAPDs. AB - The technique of random amplification of polymorphic DNA (RAPD), which is simply polymerase chain reaction (PCR) amplification of genomic DNA by a single short oligonucleotide primer, produces complex patterns of anonymous polymorphic DNA fragments. The information provided by these banding patterns has proved to be of great utility for mapping and for verification of identity of bacterial strains. Here we consider whether the degree of similarity of the banding patterns can be used to estimate nucleotide diversity and nucleotide divergence. With haploid data, fragments generated by RAPD-PCR can be treated in a fashion very similar to that for restriction-fragment data. Amplification of diploid samples, on the other hand, requires consideration of the fact that presence of a band is dominant to absence of the band. After describing a method for estimating nucleotide divergence on the basis of diploid samples, we summarize the restrictions and criteria that must be met when RAPD data are used for estimating population genetic parameters. PMID- 8412652 TI - A bias-corrected estimate of heterozygosity for single-probe multilocus DNA fingerprints. PMID- 8412653 TI - Evolutionary relationships of human populations on a global scale. AB - Using gene frequency data for 29 polymorphic loci (121 alleles), we conducted a phylogenetic analysis of 26 representative populations from around the world by using the neighbor-joining (NJ) method. We also conducted a separate analysis of 15 populations by using data for 33 polymorphic loci. These analyses have shown that the first major split of the phylogenetic tree separates Africans from non Africans and that this split occurs with a 100% bootstrap probability. The second split separates Caucasian populations from all other non-African populations, and this split is also supported by bootstrap tests. The third major split occurs between Native American populations and the Greater Asians that include East Asians (mongoloids), Pacific Islanders, and Australopapuans (native Australians and Papua New Guineans), but Australopapuans are genetically quite different from the rest of the Greater Asians. The second and third levels of population splitting are quite different from those of the phylogenetic tree obtained by Cavalli-Sforza et al. (1988), where Caucasians, Northeast Asians, and Ameridians from the Northeurasian supercluster and the rest of non-Africans form the Southeast Asian supercluster. One of the major factors that caused the difference between the two trees is that Cavalli-Sforza et al. used unweighted pair-group method with arithmetic mean (UPGMA) in phylogenetic inference, whereas we used the NJ method in which evolutionary rate is allowed to vary among different populations. Bootstrap tests have shown that the UPGMA tree receives poor statistical support whereas the NJ tree is well supported. Implications that the phylogenetic tree obtained has on the current controversy over the out-of-Africa and the multiregional theories of human origins are discussed. PMID- 8412654 TI - Molecular phylogenetics of Stenodermatini bat genera: congruence of data from nuclear and mitochondrial DNA. AB - Within the tribe Stenodermatini the systematics of the complex of species allied with the genus Artibeus has generated several alternative phylogenetic hypotheses. The most recent treatment recognized four genera (Artibeus, Dermanura, Enchisthenes, and Koopmania) and suggested that the most recent common ancestor of these four genera would include the common ancestor of all other currently recognized Stenodermatini genera except Sturnira. To test this hypothesis, we examined an EcoRI-defined nuclear satellite DNA repeat and 402 bp of DNA sequence variation from the mitochondrial cytochrome b gene. Phylogenetic conclusions based on Southern blot analyses, in situ hybridization, and mitochondrial DNA sequence data indicate that Enchisthenes is not closely related to Dermanura, Artibeus, or Koopmania and that Dermanura, Artibeus, and Koopmania shared a common ancestor after diverging from the remainder of the Stenodermatini. If our conclusions are correct, then justification for recognizing Dermanura and Koopmania as generically distinct from Artibeus must be based on the magnitude of difference that distinguishes each rather than on the conclusion that to place them as congeneric with Artibeus creates a paraphyletic taxon. PMID- 8412655 TI - Cetacean mitochondrial DNA control region: sequences of all extant baleen whales and two sperm whale species. AB - The sequence of the mitochondrial control region was determined in all 10 extant species commonly assigned to the suborder Mysticeti (baleen or whalebone whales) and to two odontocete (toothed whale) species (the sperm and the pygmy sperm whale). In the mysticetes, both the length and the sequence of the control region were very similar, with differences occurring primarily in the first approximately 160 bp of the 5' end of the L-strand of the region. There were marked differences between the mysticete and sperm whale sequences and also between the two sperm whales. The control region, less its variable portion, was used in a comparison including the 10 mysticete sequences plus the same region of an Antarctic minke whale specimen and the two sperm whales. The difference between the minke whales from the North Atlantic and the Antarctic was greater than that between any acknowledged species belonging to the same genus (Balaenoptera). The difference was similar to that between the families Balaenopteridae (rorquals) and Eschrichtiidae (gray whales). The findings suggest that the Antarctic minke whale should have a full species status, B. bonaerensis. Parsimony analysis separated the bowhead and the right whale (family Balaenidae) from all remaining mysticetes, including the pygmy right whale. The pygmy right whale is usually included in family Balaenidae. The analysis revealed a close relationship between the gray whale (family Eschrichtiidae) sequence and those of the rorquals (family Balaenopteridae). The gray whale was included in a clade together with the sei, Bryde's, fin, blue, and humpback whales. This clade was separated from the two minke whale types, which branched together. PMID- 8412656 TI - Complementation of an Enterococcus hirae (Streptococcus faecalis) mutant in the alpha subunit of the H(+)-ATPase by cloned genes from the same and different species. AB - We isolated an Enterococcus hirae (formerly Streptococcus faecalis) mutant, designated MS117, in which 'G' at position 301 of the alpha-subunit gene of the F1F0 type of H(+)-ATPase was deleted. MS117 had low H(+)-ATPase activity, was deficient in the regulatory system of cytoplasmic pH, and was unable to grow at pH 6.0. When the alpha-subunit gene of E. hirae H(+)-ATPase was ligated with the shuttle vector pHY300PLK at the downstream region of the tet gene of the vector, it was expressed without its own promoter in MS117, and the mutation of MS117 was complemented; the mutant harbouring the plasmid had the ability to maintain a neutral cytoplasm and grew at pH 6.0. We next transformed MS117 with pHY300PLK containing the alpha-subunit gene of Bacillus megaterium F1F0-ATPase constructed in the same way. The transformant grew at pH 6.0, and the ATP hydrolysis activity was recovered. These results suggested that an active hybrid H(+)-ATPase containing the B. megaterium alpha subunit was produced, and that the hybrid enzyme regulated the enterococcal cytoplasmic pH, although the function of the B. megaterium enzyme did not include pH regulation. Thus, our present results support the previous proposal that the enterococcal cytoplasmic pH is regulated by the F1F0 type of H(+)-ATPase. PMID- 8412657 TI - comF, a Bacillus subtilis late competence locus, encodes a protein similar to ATP dependent RNA/DNA helicases. AB - We have sequenced and genetically characterized comF, a Bacillus subtilis competence locus, previously identified by Tn917 transposon insertion mutagenesis. Expression of the locus, in which three open reading frames (ORFs) were found, is driven by a single sigma A-like promoter in front of comFORF1 and is dependent on early regulatory competence genes and only expressed in competence medium. The predicted amino acid sequences of two of the ORFs showed similarities to known proteins in the GenBank and SwissProt databases: ComFORF1 is similar to an extensive family of ATP-dependent RNA/DNA helicases with closer similarity to the DEAD protein subfamily and to the PriA protein in Escherichia coli. The latter is a DNA translocase/helicase required for primosome assembly at the replication fork of phage phi X174. ComFORF3 is 22% identical to Com101, a protein required for genetic competence in Haemophilus influenzae, a naturally competent Gram-negative bacterium. In-frame comFORF1 deletions were 1000-fold deficient in transformability compared to the wild-type, whereas disruptions of the other two ORFs were only five- to 10-fold lower. These observations allow us to hypothesize that the ComFORF1 late gene product plays an essential role during the binding and uptake events involved in Bacillus subtilis transformation. PMID- 8412658 TI - Conservation of motifs within the unusually variable polypeptide sequences of type I restriction and modification enzymes. AB - Type I restriction enzymes comprise three subunits encoded by genes designated hsdR, hsdM, and hsdS; S confers sequence specificity. Three families of enzymes are known and within families, but not between, hsdM and hsdR are conserved. Consequently, interfamily comparisons of M and R sequences focus on regions of putative functional significance, while both inter- and intrafamily comparisons address the origin, nature and role of diversity of type I restriction systems. We have determined the sequence of the hsdR gene for EcoA, thus making available sequences of all three hsd genes of one representative from each family. The predicted R polypeptide sequences share conserved regions with one superfamily of putative helicases, so-called 'DEAD box' proteins; these conserved sequences may be associated with the ATP-dependent translocation of DNA that precedes restriction. We also present hsdM and hsdR sequences for EcoE, a member of the same family as EcoA. The sequences of the M and R genes of EcoA and EcoE are at least as divergent as typical genes from Escherichia coli and Salmonella, perhaps as the result of selection favouring diversity of restriction specificities combined with lateral transfer among different species. PMID- 8412659 TI - Identification of nodSUIJ genes in Nod locus 1 of Azorhizobium caulinodans: evidence that nodS encodes a methyltransferase involved in Nod factor modification. AB - The Azorhizobium caulinodans strain ORS571 nodulation genes nodSUIJ were located downstream from nodABC. Complementation data and transcriptional analysis suggest that nodABCSUIJ form a single operon. Mutants with Tn5 insertions in the genes nodS, nodU, and nodJ were delayed in nodulation of Sesbania rostrata roots and stems. The NodS amino acid sequences of ORS571, Bradyrhizobium japonicum, and Rhizobium sp. strain NGR234, contain a consensus with similarity to S adenosylmethionine (SAM)-utilizing methyltransferases. A naringenin-inducible nodS-dependent protein of approximately 25 kDa could be cross-linked to radiolabelled SAM. By applying L-[methyl-3H]-methionine in vivo, Nod factors of ORS571, known to be N-methylated, could be labelled in wild type and nodU mutants but not in nodS mutants. Therefore, we propose that NodS is a SAM-utilizing methyltransferase involved in Nod factor synthesis. PMID- 8412660 TI - Co-dependent positive regulation of the ansB promoter of Escherichia coli by CRP and the FNR protein: a molecular analysis. AB - Transcription of the ansB gene, encoding L-asparaginase II, is positively regulated by cAMP receptor protein (CRP) and by the product of the fnr gene, the FNR protein. These global regulatory proteins mediate the expression of ansB in Escherichia coli in response to carbon source and to anaerobiosis, respectively, and are required concurrently for optimal ansB expression. The mechanism whereby CRP and FNR interact co-operatively with the ansB promoter to achieve transcription has not previously been established. We have utilized an ansB' 'lacZ fusion, in conjunction with deletion analysis and site-directed mutagenesis, to identify two sites which interact with these regulatory proteins in the ansB promoter. The first is an FNR site, centred 41.5 bp upstream of the major transcriptional start site. The second site, located 28 bp upstream of the FNR site, is the site of CRP regulation. This site is homologous to both the CRP and FNR binding-site consensus sequences and may respond to both CRP and FNR. The concurrent requirement for CRP and FNR for optimal expression of ansB may be explained if, first, essentially no transcription occurs unless the FNR is bound at the downstream site, and, second, the level of transcription when FNR alone is present is enhanced when CRP binds at the upstream site. PMID- 8412661 TI - Regulation of the ansB gene of Salmonella enterica. AB - The expression of L-asparaginase II (encoded by ansB) in Salmonella enterica was found to be positively regulated by the cAMP receptor protein (CRP) and anaerobiosis. The anaerobic regulation of the S. enterica ansB gene is not mediated by the anaerobic transcriptional activator FNR. This is unlike the situation of the ansB gene of Escherichia coli, which is dependent on both CRP and FNR. To investigate this fundamental difference in the regulation of L asparaginase II expression in S. enterica, the ansB gene was cloned and the nucleotide sequence of the promoter region determined. Sequence analysis and transcript mapping of the 5' promoter region revealed a single transcriptional start point (tsp) and two regulatory sites with substantial homology with those found in E. coli. One site, centred -90.5 bp from the tsp, is homologous to a hybrid CRP/FNR ('CF') site which is the site of CRP regulation in the E. coli promoter. The other site, centred 40.5 bp upstream of the tsp, is homologous to the FNR binding site of the E. coli promoter. Significantly, however, a single base-pair difference exists in this site, at a position of the related CRP and FNR DNA-binding site consensus sequences known to be involved in CRP versus FNR specificity. Site-directed mutagenesis indicates that this single difference, relative to the homologous E. coli site, results in a CRP binding site and the observed FNR-independent ansB expression in S. enterica.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412662 TI - Molecular cloning and characterization of a sym plasmid locus that regulates cultivar-specific nodulation of soybean by Rhizobium fredii USDA257. AB - Rhizobium fredii strain USDA257 produces nitrogen-fixing nodules on primitive soybean cultivars such as Peking but fails to nodulate agronomically improved cultivars such as McCall. Transposon-mutant 257DH4 has two new phenotypes: it nodulates McCall, and its ability to do so is sensitive to the presence of parental strain USDA257, i.e. it is subject to competitive nodulation blocking. We have isolated a cosmid containing DNA that corresponds to the site of transposon insertion in 257DH4 and have localized Tn5 on an 8.0 kb EcoRI fragment. The 5596 bp DNA sequence that surrounds the insertion site contains seven open reading frames. Five of these, designated nolBTU, ORF4, and nolV, are closely spaced and of the same polarity. nolW and nolX are of the opposite polarity. The initiation codon for nolW lies 155 bp upstream from that of nolB, and its is separated from nolX by 281 bp. The predicted NolT and NolW proteins have putative membrane-spanning regions. The N-terminus of the hypothetical NolW protein also has limited homology to NodH of Rhizobium meliloti, but none of the deduced protein sequences has significant homology to known nodulation gene products. Site-directed mutagenesis with mudII1734 confirms that inactivation of nolB, nolT, nolU, nolV, nolW, or nolX extends host range for nodulation to McCall soybean. This phenotype could not be genetically dissected from sensitivity to competitive nodulation blocking. Expression of nolBTU and nolX is induced as much as 30-fold by flavonoid signal molecules, even though these genes lack nod-box promoters. Histochemical staining of McCall roots inoculated with nolB-, nolU-, or nolX-lacZ fusions verifies that these genes are expressed continuously from preinfection to the stage of the functional nodule. Although a nolU-ORF4-nolV clone hybridizes to a single 8.0 kb EcoRI fragment from 10 strains of R. fredii and broad-host-range Rhizobium sp. NGR234, hybridizing sequences are not detectable in other rhizobia. PMID- 8412663 TI - Analysis of diphtheria toxin repressor-operator interactions and characterization of a mutant repressor with decreased binding activity for divalent metals. AB - The diphtheria toxin repressor (DtxR) is an Fe(2+)-activated protein with sequence-specific DNA-binding activity for the diphtheria toxin (tox) operator. Under high-iron conditions in Corynebacterium diphtheriae, DtxR represses toxin and siderophore biosynthesis as well as iron uptake. DtxR and a mutant repressor with His-47 substituted for Arg-47, designated DtxR-R47H, were purified and compared. Six different divalent cations (Cd2+, Co2+, Fe2+, Mn2+, Ni2+, and Zn2+) activated the sequence-specific DNA-binding activity of DtxR and enabled it to protect the tox operator from DNase I digestion, but Cu2+ failed to activate DtxR. Hydroxyl radical footprinting experiments indicated that DtxR binds symmetrically about the dyad axis of the tox operator. Methylation protection experiments demonstrated that DtxR binding alters the susceptibility to methylation of three G residues within the AT-rich tox operator. These findings suggest that two or more monomers of DtxR are involved in binding to the tox operator, with symmetrical DNA-protein interactions occurring at each end of the palindromic operator. In this regard, DtxR resembles several other well characterized prokaryotic repressor proteins but differs dramatically from the Fe(2+)-activated ferric uptake repressor protein (Fur) of Escherichia coli. The concentration of Co2+ required to activate DtxR-R47H was at least 10-fold greater than that needed to activate DtxR, but the sequence-specific DNA binding of activated DtxR-R47H was indistinguishable from that of wild-type DtxR. The markedly deficient repressor activity of DtxR-R47H is consistent with a significant decrease in its binding activity for divalent cations. PMID- 8412664 TI - Cloning and characterization of the psaE gene of the cyanobacterium Synechococcus sp. PCC 7002: characterization of a psaE mutant and overproduction of the protein in Escherichia coli. AB - The psaE gene, encoding a 7.5 kDa peripheral protein of the photosystem I complex, has been cloned and characterized from the cyanobacterium Synechococcus sp. PCC 7002. The gene is transcribed as an abundant monocistronic transcript of approximately 325 nt. The PsaE protein has been overproduced in Escherichia coli, purified to homogeneity, and used to raise polyclonal antibodies. Mutant strains, in which the psaE gene was insertionally inactivated by interposon mutagenesis, were constructed and characterized. Although the PS I complexes of these strains were similar to those of the wild type, the strains grew more slowly under conditions which favour cyclic electron transport and could not grow at all under photoheterotrophic conditions. The results suggest that PsaE plays a role in cyclic electron transport in cyanobacteria. PMID- 8412665 TI - Molecular analysis of two ScrR repressors and of a ScrR-FruR hybrid repressor for sucrose and D-fructose specific regulons from enteric bacteria. AB - The scr regulon of pUR400 and the chromosomally encoded scr regulon of Klebsiella pneumoniae KAY2026 are both negatively controlled by a specific repressor (ScrR). As deduced from the nucleotide sequences, both scrR genes encode polypeptides of 334 residues (85.5% identical base pairs, 91.3% identical amino acids), containing an N-terminal helix-turn-helix motif. Comparison with other regulatory proteins revealed 30.6% identical amino acids to FruR, 27.0% to Lacl and 28.1% to GalR. Six scrRs super-repressor mutations define the inducer-binding domain. The scr operator sequences were identified by in vivo titration tests of the sucrose repressor and by in vitro electrophoretic mobility shift assays. D-fructose, an intracellular product of sucrose transport and hydrolysis, and D-fructose 1 phosphate were shown to be molecular inducers of both scr regulons. An active ScrR-FruR hybrid repressor protein was constructed with the N-terminal part of the sucrose repressor of K. pneumoniae and the C-terminal part of the fructose repressor of Salmonella typhimurium LT2. Gel retardation assays showed that the hybrid protein bound to scr-specific operators, and that D-fructose 1-phosphate, the inducer for FruR, was the only inducer. In vivo, neither the operators of the fru operon nor of the pps operon, the natural targets for FruR, were recognized, but the scr operators were. These data and the data obtained from the super repressor alleles confirm previous models on the binding of repressors of the Lacl family to their operators. PMID- 8412666 TI - A family of IS1031 elements in the genome of Acetobacter xylinum: nucleotide sequences and strain distribution. AB - An insertion sequence (here called IS1031A) from Acetobacter xylinum ATCC 23769 has recently been isolated. This study describes the complete nucleotide sequence of IS1031A as well as the sequences of two novel iso-IS1031 elements, IS1031C and IS1031D, from A. xylinum ATCC 23769. The three ISs are all exactly 930 bp long, have imperfect terminal inverted repeats of 24 bp for IS1031A and 21 bp for IS1031C and IS1031D, are flanked by three base pair direct repeats, and contain an open reading frame encoding a putative basic protein of 278 amino acids. Because of nucleotide substitutions, IS1031C and IS1031D differ from IS1031A by 12.9% while IS1031C differs from IS1031D by only 0.6%. Hybridization analyses of total DNA from nine A. xylinum strains showed that all strains contained IS1031 like elements varying in copy number from three to at least 16. None of three Acetobacter aceti strains examined contained IS1031-like elements. Taken together, the results suggest that A. xylinum contains a family of IS1031 elements with considerably diversified nucleotide sequences. PMID- 8412667 TI - The molecular interaction between components of the TonB-ExbBD-dependent and of the TolQRA-dependent bacterial uptake systems. PMID- 8412668 TI - The general amino acid control regulates MET4, which encodes a methionine-pathway specific transcriptional activator of Saccharomyces cerevisiae. PMID- 8412669 TI - Regulation of epidermin biosynthetic genes by EpiQ. AB - We investigated the role of epiQ in the biosynthesis of the lantibiotic epidermin. epiQ was essential for epidermin production. It was shown that EpiQ controls epidermin production by transcriptionally activating the epiA promoter, used for transcription of most of the epidermin biosynthetic genes. Additional copies of epiQ increased epidermin production in the epidermin-producing wild type strain Staphylococcus epidermidis Tu3298. The epiA promoter region was characterized by primer extension analysis. Two inverted repeats, putative operator sites for EpiQ binding, are located upstream of the -35 region and one is localized downstream of the -10 region. Crude protein extracts from S. epidermidis Tu3298 and epiQ expressing Escherichia coli cells led to gel mobility shifts of a DNA fragment bearing the inverted repeat which is located immediately upstream of the -35 region. DNA fragments bearing the other two inverted repeats were not shifted. The epiQ gene product could be detected by overexpression in the E. coli T7 system using antiserum raised against synthetic peptides of EpiQ. Furthermore, EpiQ, like other DNA-binding proteins, was shown to bind strongly to heparin sepharose. PMID- 8412670 TI - Site-specific insertion of gene cassettes into integrons. AB - Site-specific insertion of gene cassettes into the insert region of integrons has been demonstrated. Insertion was only observed if the integron DNA integrase was expressed in the recipient cell and if the cassette DNA was ligated prior to transformation. The essential ligation products were resistant to treatment with exonuclease III, indicating that they were closed circular molecules. Insertion of cassettes into integron fragments containing either no insert (one recombination site), or one gene cassette (two recombination sites), was demonstrated. In the latter case, insertion occurred predominantly at the core site located 5' to the resident cassette, which corresponds to the only site available when no insert is present in the recipient. When DNA molecules including two gene cassettes were used, insertion of only one of the gene cassettes was generally observed, suggesting that resolution of the circular molecule to generate two independent circular cassettes occurred more rapidly than insertion into the recipient integron. PMID- 8412671 TI - Iron and oxygen regulation of Escherichia coli MnSOD expression: competition between the global regulators Fur and ArcA for binding to DNA. AB - Manganese superoxide dismutase (the sodA gene product) in Escherichia coli, is negatively regulated by two global regulators, ArcA (aerobic respiration control) and Fur (ferric uptake regulation), coupling its expression to aerobic metabolism and the intracellular iron pool. Footprinting analyses were carried out on the sodA promoter region with purified Fur protein and with ArcA protein overproduced in crude extracts. ArcA is able to bind in vitro in the absence of the in vivo triggering signal. The binding occurs in one step and study of contacts within the operator sequence reveals binding on one side of the double helix. The DNA protection extends to a much larger region (about 65 bp) than would be expected for a 27 kDa protein, suggesting polymerization. Both Fur and ArcA footprints encompass the -35 and -10 promoter region and there is considerable overlap of their binding sequences, in agreement with in vivo results suggesting that either regulator alone can block sodA transcription. Furthermore, competition experiments show that Fur and ArcA binding to the sodA promoter are mutually exclusive and that ArcA can easily displace Fur, but not vice versa. The biological significance of this in vitro behaviour is discussed. PMID- 8412672 TI - S-layer protein from Thermus thermophilus HB8 assembles into porin-like structures. AB - The cells of the extreme thermophile Thermus thermophilus are surrounded by a regular layer (S-layer) built up by a protein with an apparent molecular mass of 100 kDa (P100). From purified membrane fractions, three different class of two dimensional crystals can be obtained by following alternative extractive procedures. One of these crystals, with p6 symmetry, clearly represents the native S-layer detected by freeze etching on whole cells, while the other two, showing p2 and p3 symmetries respectively, closely resemble aggregates of bacterial porins. We demonstrate here by limited proteolysis and Western blotting the surprising fact that the protein component of the three crystals is the P100 protein. Our biochemical data also show how this protein forms Ca(2+)-stabilized trimers in each crystal, which support the structural analysis that points to p3 units as the common structural block in all of them, and again resembles the situation found in bacterial porins. PMID- 8412673 TI - Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena. AB - The spatially patterned differentiation of heterocysts in the filamentous cyanobacterium Anabaena requires a functional hetR gene. Transcriptional fusions to luxAB show that hetR is transcribed at a low level throughout the filament when Anabaena is grown with combined nitrogen, and that induction of the gene begins within 2 h following nitrogen deprivation. By 3.5 h, induction is localized to spaced foci. By 6 h, there is an overall induction of at least threefold in whole cultures, reflecting at least a 20-fold increase within spatially separated cells. The induction requires the presence of a functional hetR gene, indicating that hetR is autoregulatory. Full induction of a heterocyst structural gene, hepA, also requires a functional hetR locus. PMID- 8412674 TI - Cloning and nucleotide sequence of frpC, a second gene from Neisseria meningitidis encoding a protein similar to RTX cytotoxins. AB - Neisseria meningitidis FAM20 has recently been shown to produce two Fe-regulated proteins (FrpA and FrpC) related to the RTX family of cytotoxins. Here we report the cloning and DNA sequence of the locus containing the gene encoding the larger meningococcal RTX protein FrpC. FrpC was highly similar to FrpA throughout much of the predicted protein, with two main differences. Whereas the FrpA protein had 13 copies of the nine-amino-acid repeat units typical of RTX proteins, FrpC had 43 copies. The additional copies in FrpC apparently arose from a threefold tandem amplification of a 600bp DNA fragment encoding the repeats. In addition, the frpC gene lacked good promoter consensus sequences. An open reading frame (ORF1) of unknown function was found immediately upstream of frpC, suggesting the possibility that frpC was cotranscribed with ORF1. A probable promoter was found 300bp upstream of ORF1, and it contained a Fur protein-binding sequence found in the promoters of Fe-regulated Escherichia coli genes. DNA upstream of the ORF1/frpC promoter was homologous to IS1016-like elements surrounding capsulation loci of strains of Haemophilus influenzae. A FrpC-like protein (reactive in immunoblots with monoclonal antibody 9D4; multiple reactive bands of about 200 to 120 kDa) was found in five out of eight meningococcal strains but only in one out of 14 other Neisseria, suggesting that FrpC may participate in the pathogenesis of meningococcal disease. PMID- 8412675 TI - Adaptation of Escherichia coli to redox environments by gene expression. AB - Escherichia coli is adroit in exploiting environmental energy sources to its greatest profit. A key strategy is to channel electron transport from donor to a terminal acceptor(s) so that the voltage drop is maximal. At the level of transcription, the goal is achieved by the interaction of three global regulatory systems, Fnr, NarL/NarX and ArcB/ArcA. In addition, the regulator FhlA is involved in a cascade-controlled pathway for the formate branch of the pyruvate fermentation pathway. PMID- 8412676 TI - Characterization of the cspB gene encoding PS2, an ordered surface-layer protein in Corynebacterium glutamicum. AB - PS2 is one of two major proteins detected in the culture media of various Corynebacterium glutamicum strains. The coding and promoter regions of the cspB gene encoding PS2 were cloned in lambda gt11 using polyclonal antibodies raised against PS2 for screening. Expression of the cspB gene in Escherichia coli led to the production of a major anti-PS2 labelled peptide of 63,000 Da, corresponding presumably to the mature form of PS2. It was detected in the cytoplasm, periplasm and surrounding medium of E. coli. Three other slower migrating bands of 65,000 68,000 and 72,000 Da were detected. The largest one probably corresponds to the precursor form of PS2 in E. coli. Analysis of the nucleotide sequence revealed an open reading frame (ORF) of 1533 nucleotides. The deduced 510-amino-acid polypeptide had a calculated molecular mass of 55,426 Da. According to the predicted amino acid sequence, PS2 is synthesized with a N-terminal segment of 30 amino-acid residues reminiscent of eukaryotic and prokaryotic signal peptides, and a hydrophobic domain of 21 residues near the C-terminus. Although no significant homologies were found with other proteins, it appears that some characteristics and the amino acid composition of PS2 share several common features with surface-layer proteins. The cspB gene was then disrupted in C. glutamicum by gene replacement. Freeze-etching electron microscopy performed on the wild-type strain indicated that the cell wall of C. glutamicum is covered with an ordered surface of proteins (surface layer, S-layer) which is in very close contact with other cell-wall components. These structures are absent from the cspB-disrupted strain but are present after reintroduction of the cspB gene on a plasmid into this mutant. Thus we demonstrate that the S-layer protein is the product of the cspB gene. PMID- 8412677 TI - Structure-function correlation for the EcoRV restriction enzyme: from non specific binding to specific DNA cleavage. AB - The EcoRV restriction endonuclease cleaves DNA at its recognition sequence at least a million times faster than at any other DNA sequence. The only cofactor it requires for activity is Mg2+; but in binding to DNA in the absence of Mg2+, the EcoRV enzyme shows no specificity for its recognition site. Instead, the reason why EcoRV cuts one DNA sequence faster than any other is that the rate of cleavage is controlled by the binding of Mg2+ to EcoRV-DNA complexes: the complex at the recognition site has a high affinity for Mg2+, while the complexes at other DNA sequences have low affinities for Mg2+. The structures of the EcoRV endonuclease, and of its complexes with either specific or non-specific DNA, have been solved by X-ray crystallography. In the specific complex, the protein interacts with the bases in the recognition sequence and the DNA takes up a highly distorted structure. In the non-specific complex with an unrelated DNA sequence, there are virtually no interactions with the bases and the DNA retains a B-like structure. Since the free energy changes for the formation of specific and non-specific complexes are the same, the energy from the specific interactions balances that required for the distortion of the DNA. The distortion inserts the phosphate at the scissile bond into the active site of the enzyme, where it forms part of the binding site for Mg2+. Without this distortion, the EcoRV-DNA complex would be unable to bind Mg2+ and thus unable to cleave DNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412678 TI - Preferential cis action of IS10 transposase depends upon its mode of synthesis. AB - A number of bacterial DNA-binding proteins, including IS element transposases, act preferentially in cis. We show below that the degree of preferential cis action by IS10 transposase depends upon its mode of synthesis at steps subsequent to transcription initiation. Cis preference is increased several fold by mutations that decrease translation initiation, by the presence of IS10-specific antisense RNA and by plasmids that increase the level of cellular RNases. Conversely, cis preference is decreased by mutations that increase translation initiation; in some cases, cis preference is nearly abolished. Mutations that alter the rate of transcription initiation have no effect. In light of other observations, we suggest that cis preference is strongly dependent upon the rate at which transcripts are released from their templates and/or the half-life of the transposase message. These observations provide further evidence that inefficient translation plays multiple roles in the biology of IS10. PMID- 8412679 TI - The int genes of bacteriophages P22 and lambda are regulated by different mechanisms. AB - Bacteriophage P22 and lambda are related bacteriophages with similar gene organizations. In lambda the cll-dependent Pl promoter is responsible for lambda int gene expression. The only apparent counterpart to pl in P22 is oriented in the opposite direction, and cannot transcribe the P22 int gene. We show that this promoter, called P(al), is active both in vivo and in vitro, and is dependent upon the P22 cll-like gene, called c1. We have also determined the DNA sequence of a 3.3 kb segment that closes the gap between previously reported sequences to give a continuous sequence between the P22 pL promoter and the int gene. The newly determined sequence is densely packed with genes from the pL direction, and the proteins predicted by the sequence show excellent correlation with the proteins mapped by Youderian and Susskind in 1980. However, the sequence contains no apparent genes in the opposite (p(al)) direction, and no additional binding motifs for the P22 c1 protein. We conclude that int gene expression in P22 is regulated by a different mechanism than in lambda. PMID- 8412680 TI - Hc1-mediated effects on DNA structure: a potential regulator of chlamydial development. AB - Chlamydiae are obligate intracellular bacteria which undergo a unique developmental cycle, alternating between non-replicative elementary bodies (EBs) and replicative reticulate bodies (RBs). The transition from RB to EB is characterized by condensation of the chromosome into a dense nucleoid structure. The chlamydial histone homologue Hc1 is sufficient to induce formation of a similar structure in Escherichia coli. High-level Hc1 expression in E. coli is self-limiting and down-regulates transcription, translation, and replication at concentrations similar to those observed in chlamydial elementary bodies. Expression of Hc1 at sub-structural levels may have specific regulatory functions through its interaction with chromosomal DNA. In E. coli this is reflected in a dramatic shift in the pattern of gene expression. The differential expression of the outer membrane porin proteins OmpC and OmpF and analysis of lacZ fusions with promoter regions sensitive to supercoiling suggests that low-level Hc1 expression results in a net relaxation of chromosomal DNA. Topological analysis of plasmid DNA from both E. coli and Chlamydia trachomatis supports a decrease in superhelicity preceding nucleoid formation. In vitro analysis of purified Hc1-DNA interactions supports preferential binding based upon DNA conformation. These results suggest a dual role in which Hc1-mediated changes in gene expression may precede metabolic inactivity. PMID- 8412681 TI - Pleiotropic effects of the rpoC10 mutation affecting the RNA polymerase beta' subunit of Escherichia coli on factor-dependent transcription termination and antitermination. AB - Escherichia coli RNA polymerase is composed of four different subunits, 2 alpha, beta, beta' and sigma. Among these subunits, the role of beta' is poorly understood. The rpoC10 mutation affecting beta' has been isolated as a suppressor mutation of the temperature-sensitive nusA11 mutant. DNA sequence analysis revealed that the rpoC10 mutant is a substitution of Lys for Glu-402. This increased positive charge appears to compensate for the increased negative charge present in the nusA11 protein (Asp for Gly-181). In vivo measurements of reporter gene expression have revealed that rpoC10 restores rho-dependent termination but fails to restore rho-independent termination in nusA11. Moreover, the rpoC10 mutation, in combination with any nusA mutation, inhibited lambda Q-mediated antitermination without affecting N antitermination and severely restricted lambda phage development. The inhibition of Q function and lambda growth could be compensated for by overproducing Q. These results suggest that the RNA polymerase beta' subunit plays a crucial role in factor-dependent transcription termination and antitermination. PMID- 8412682 TI - Processing and methylation of PuIG, a pilin-like component of the general secretory pathway of Klebsiella oxytoca. AB - The signal sequence of the Klebsiella oxytoca puIG gene product, which is required for extracellular secretion of the enzyme pullulanase, is similar in many respects to the corresponding segment of the precursors of type IV (me-Phe) pilins. The significance of this similarity is confirmed by the observation that the puIO gene product processes prePuIG at the consensus type IV prepilin peptidase cleavage site at the amino-terminal end of the PuIG signal sequence. Like most type IV pilins, processed PuIG was found to have a methylated amino terminal phenylalanine residue. Site-directed mutagenesis was used to replace amino acids in prePuIG that correspond to residues shown by others to be essential for processing, methylation and assembly of type IV pilins. The glycine residue on the amino-terminal side of the prePuIG cleavage site is absolutely required for processing and for pullulanase secretion. The glutamate residue at position 11(+5) is also required for pullulanase secretion but not for processing or methylation. This result contrasts with that reported for corresponding variants of Pseudomonas aeruginosa type IV prepilin, which were processed but only inefficiently N-methylated. Cleavage of prePuIG and pullulanase secretion were both unaffected by replacement of the phenylalanine residue on the carboxy terminal side of the cleavage site by leucine, isoleucine or valine, by a conservative substitution within the hydrophobic core of the prePuIG signal sequence, or by a glutamine to proline substitution within the processed segment. However, replacement of the same glutamine residue by arginine abolished secretion without affecting either processing or methylation. PMID- 8412683 TI - Cloning, expression and sequencing of Helicobacter felis urease genes. AB - Urease genes from Helicobacter felis were cloned and expressed in Escherichia coli cells. A genomic bank of Sau3A-digested H. felis chromosomal DNA was created using a cosmid vector. Cosmid clones were screened for urease activity following subculture on a nitrogen-limiting medium. Subcloning of DNA from an urease positive cosmid clone led to the construction of pILL205 (9.5 kb) which conferred a urease activity of 1.2 +/- 0.5 mumole urea min-1 mg-1 bacterial protein to E. coli HB101 bacteria grown on a nitrogen-limiting medium. Random mutagenesis using a MiniTn3-Km transposable element permitted the identification of three DNA regions on pILL205 which were necessary for the expression of an urease-positive phenotype in E. coli clones. To localize the putative structural genes of H. felis on pILL205, extracts of clones harbouring the mutated copies of the plasmid were analysed by Western blotting with anti-H. felis rabbit serum. One mutant clone did not synthesize the putative UreB subunit of H. felis urease and it was postulated that the transposable element had disrupted the corresponding structural gene. By sequencing the DNA region adjacent to the transposon insertion site two open reading frames, designated ureA and ureB, were identified. The polypeptides encoded by these genes had calculated molecular masses of 26,074 and 61,663 Da, respectively, and shared 73.5% and 88.2% identity with the corresponding gene products of Helicobacter pylori urease. PMID- 8412684 TI - Double, independent mutational events in the rpsL gene of Escherichia coli: an example of hypermutability? AB - A proportion (up to 20%) of newly arising streptomycin-dependent (SmD) colonies of Escherichia coli WP2 contain bacteria where, in addition to a known SmD determining (primary) mutation in the rpsL gene, there is a further ancillary mutation in the same gene. These ancillary mutations occur at 10 sites between 11 and 201bp away from the primary mutation. Ancillary mutations have been found in mutant colonies arising both spontaneously and after treatment with ultraviolet light and some have been found repeatedly. Ancillary mutations were frequently found to occur in mixed clones with an excess of bacteria carrying only the primary SmD mutation. No ancillary mutations were found in an adjacent non-coding region and there were no coding sequence changes that did not alter the amino acid specified. Although a selective advantage for bacteria containing ancillary mutations could not be demonstrated directly in every case, some small advantage must be presumed to have occurred to explain the absence of mutations at the other sites and particularly at third (wobble) codon positions. Ancillary mutations appear to occur fairly early in the life of a newly arisen SmD mutant clone in some sort of hypermutable process. Whether they are noticed appears to depend on their conferment of some selective advantage on the bacteria carrying them. While the ancillary mutations within rpsL lie close to, and may be consequent upon the formation of a primary SmD mutation, their mechanism of formation appears to be at least to some extent independent and does not involve the recA or umuC genes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412685 TI - A pleiotropic reduced virulence (Rvi-) mutant of Erwinia carotovora subspecies atroseptica is defective in flagella assembly proteins that are conserved in plant and animal bacterial pathogens. AB - Erwinia carotovora subsp. atroseptica was mutagenized and assayed for virulence in planta. Those mutants which exhibited reduced virulence (Rvi-) were assayed for growth rate, auxotrophy and extracellular enzyme secretion and seven mutants were found to be wild type for all of these phenotypes. When screened for other phenotypes, two were found to be non-motile. One mutant was complemented for motility by a heterologous gene library. A 2.7kb XmaIII-ClaI complementing fragment was sequenced and the gene products were found to have similarity to flagella biosynthesis gene products from several bacteria. Further similarity was found to a pathogenicity protein from the plant pathogen Xanthomonas campestris pv. glycines and to the Spa pathogenicity proteins of the human pathogen Shigella flexneri, which are involved in the surface presentation of antigens. These studies highlight the emergence of common themes in the molecular strategies employed by both plant and animal bacterial pathogens for the targeting of proteins involved in the elaboration of disease. PMID- 8412686 TI - Generation of isogenic K54 capsule-deficient Escherichia coli strains through TnphoA-mediated gene disruption. AB - Transposon mutagenesis, using IS50L::phoA(Tn-phoA), was performed in a K54/O4/H5 blood isolate of Escherichia coli (CP9), to generate a library of random mutants. Five hundred and twenty-six independent CP9 TnphoA mutants were isolated with active gene fusions to alkaline phosphatase. From this mutant library, eight capsule-deficient strains were detected and were found to have a single copy of TnphoA. Sixteen additional capsule deficient mutants with TnphoA inserts were subsequently obtained that did not possess active PhoA fusions. In conjunction with the initial eight capsule-deficient isolates we have defined genes on three different XbaI fragments as being involved in capsule production. Generalized transduction with the bacteriophage T4 established that these insertions were responsible for the loss of capsule and that they are linked. These capsule deficient strains can be used to assess the pathogenic role of the K54 capsular polysaccharide. PMID- 8412687 TI - Sequence and properties of mecA, a negative regulator of genetic competence in Bacillus subtilis. AB - The development of competence in Bacillus subtilis is regulated by growth conditions and several regulatory genes. In complex media competence development is poor, and there is little or no expression of late competence genes. mec mutations permit competence development and late competence gene expression in complex media, and bypass the requirements for many of the competence regulatory genes. In this paper we describe the cloning and characterization of mecA. The mecA gene product acts negatively in the development of competence. Null mutations in mecA allowed expression of a late competence gene comG, under conditions where it is not normally expressed, including in complex media and in cells mutant for several competence regulatory genes. Overexpression of MecA from a multicopy plasmid resulted in inhibition of comG transcription. The DNA sequence of mecA was determined and the predicted gene product showed no significant similarity to any protein in the database. Expression of a mecA-lacZ translational fusion was constitutive during growth and did not vary significantly in the different media tested. The role of mecA in competence development and other stationary phase phenomena is discussed. PMID- 8412688 TI - Isolation and characterization of hydroxylamine-induced mutations in the Erwinia herbicola ice nucleation gene that selectively reduce warm temperature ice nucleation activity. AB - Cells of ice nucleation active bacterial species catalyse ice formation over the temperature range of -2 to -12 degrees C. Current models of ice nucleus structure associate the size of ice nucleation protein aggregates with the temperature at which they catalyse ice formation. To better define the structural features of ice nucleation proteins responsible for the functional heterogeneity of ice nuclei within a genetically homogeneous collection of cells we used in vitro chemical mutagenesis to isolate mutants with reduced ability to nucleate ice at warm assay temperatures but which retain normal or near normal nucleation activity at cold temperatures (WIND, i.e. warm ice nucleus-deficient mutants). Nearly half of the mutants obtained after hydroxylamine mutagenesis of the iceE gene from Erwinia herbicola had this phenotype. The phenotypes and location of lesions on the genetic map of iceE were determined for a number of mutants. All WIND mutations were restricted to the portion of iceE encoding the repetitive region of the polypeptide. DNA sequencing of two WIND mutants revealed single nucleotide substitutions changing a conserved serine or glycine residue to phenylalanine and serine, respectively. The implications of these findings in structure/function models for the ice nucleation protein are discussed. PMID- 8412689 TI - FtsZ ring in bacterial cytokinesis. AB - FtsZ is localized to a cytokinetic ring at the cell division site in bacteria. In this review a model is discussed that suggests that FtsZ self assembles into a ring at a nucleation site formed on the cytoplasmic membrane under cell-cycle control. This model suggests that formation of the cytokinetic FtsZ ring initiates and coordinates the circumferential invagination of the cytoplasmic membrane and cell wall, leading to formation of the septum. It is also suggested that this process may be conserved among the peptidoglycan-containing eubacteria. In addition, similarities between FtsZ and tubulin are discussed. PMID- 8412690 TI - Translation elongation factor 3: a fungus-specific translation factor? AB - Fungi appear to be unique in their requirement for a third soluble translation elongation factor. This factor, designated elongation factor 3 (EF-3), was first described in the yeast Saccharomyces cerevisiae and has subsequently been identified in a wide range of fungal species including Candida albicans and Schizosaccharomyces pombe. EF-3 exhibits ribosome-dependent ATPase and GTPase activities that are not intrinsic to the fungal ribosome, but which are essential for translation elongation. Recent studies on the structure of EF-3 from several fungal species have shown that it consists of a repeated domain, with each domain containing the expected putative ATP- and GTP-binding motifs. Overall, EF-3 shows striking amino acid similarity to members of the ATP-binding Cassette (ABC) family of membrane-associated transport proteins although EF-3 is not itself directly membrane-associated. Regions of the EF-3 polypeptide also show structural homology with other translation-associated factors including aminoacyl tRNA synthetases and the Escherichia coli ribosomal protein S5. While the precise role of EF-3 in the translation elongation cycle remains to be defined, recent evidence suggests that it may be involved in optimizing accuracy during mRNA decoding at the ribosomal A site. Furthermore, the essential nature of EF-3 with respect to the fungal cell indicates that it may be an effective antifungal target. Its apparently ubiquitous occurrence throughout the fungal kingdom also suggests that it may be a useful fungal taxonomic marker. PMID- 8412691 TI - Assembly of the Escherichia coli F1F0 ATPase, a large multimeric membrane-bound enzyme. AB - The F1F0 proton translocating ATPase of Escherichia coli is a large membrane bound enzyme complex consisting of more than 20 polypeptides that are encoded by the unc operon. Besides being a system for analysing the enzymology of ATP synthesis and energy coupling, the ATPase is a model system for determining how large oligomeric membrane-bound proteins are synthesized and assembled. The assembly of the ATPase involves differential gene expression and assembly of the subunits within the membrane and with each other. This review discusses the influence of F1 subunits on the assembly and proton permeability of the F0 proton channel, and the possible advantages to assembly of the particular arrangement of genes in the unc operon. PMID- 8412692 TI - Nitrate regulation of anaerobic respiratory gene expression in Escherichia coli. AB - Synthesis of most anaerobic respiratory pathways is subject to dual regulation by anaerobiosis and nitrate. Anaerobic induction is mediated by the FNR protein. Dual interacting two-component regulatory systems mediate nitrate induction and repression. The response regulator protein NARL binds DNA to control nitrate induction and repression of genes encoding nitrate respiration enzymes and alternate anaerobic respiratory enzymes, respectively. The homologous protein NARP controls nitrite induction of at least two operons. Nitrate and nitrite signalling are both mediated by the homologous sensor proteins NARX and NARQ. Recent mutational analyses have defined a heptamer sequence necessary for specific DNA binding by the NARL protein. These heptamers are located at different positions in the control regions of different operons. The NARL protein binding sites in the narG (nitrate reductase) and narK (nitrate-nitrite antiporter) operon control regions are located approximately 200bp upstream of the transcription initiation site. The integration host factor (IHF) greatly stimulates nitrate induction of these operons, indicating that a specific DNA loop brings NARL protein, bound at the upstream region, into the proximity of the promoter for transcription activation. Other NARL protein-dependent opersons do not require IHF for nitrate induction, and the arrangement of NARL heptamer sequences in these control regions is quite different. This complexity of signal transduction pathways, coupled with the diversity of control region architecture, combine to provide many interesting areas for future investigation. An additional challenge is to determine how or if the FNR and NARL proteins interact to mediate dual positive control of transcription initiation. PMID- 8412693 TI - The FtsZ protein of Bacillus subtilis is localized at the division site and has GTPase activity that is dependent upon FtsZ concentration. AB - The ftsZ gene is essential for cell division in both Escherichia coli and Bacillus subtilis. In E. coli FtsZ forms a cytokinetic ring at the division site whose formation is under cell-cycle control. In addition, the FtsZ from E. coli has a GTPase activity that shows an unusual lag in vitro. In this study we show that FtsZ in Bacillus subtilis forms a ring that is at the tip of the invaginating septum. The FtsZ ring is dynamic since it is formed as division is initiated, changes diameter during septation, and disperses upon completion of septation. In vitro the purified FtsZ from B. subtilis exhibits a GTPase activity without a demonstrable lag, but the GTPase activity is markedly dependent upon the FtsZ concentration, suggesting that the FtsZ protein must oligomerize to express the GTPase activity. PMID- 8412694 TI - The genes of the glutamine synthetase adenylylation cascade are not regulated by nitrogen in Escherichia coli. AB - Regulation of glutamine-synthetase (GS) activity in enteric bacteria involves a complex cascade of events. In response to nitrogen limitation, a transferase catalyses the uridylylation of the PII protein, which in turn stimulates deadenylylation of GS. Deadenylylated GS is the more active form of the enzyme. Here we characterize in detail the genes from Escherichia coli encoding uridylyl transferase (glnD), the PII protein (glnB), and adenylyl-transferase (glnE). glnD is transcribed from its own promoter, glnE is contranscribed with another gene, orfXE, whereas glnB is partly contranscribed with a gene encoding a homologue of the transcription activator NtrC. All three gln regulatory genes were constitutively expressed at a low level, i.e. their expression was independent of the nitrogen status and the RNA polymerase sigma factor sigma 54. We conclude that the functioning of the GS adenylylation cascade is regulated by modulation of the activities of uridylyl-transferase and adenylyl-transferase, rather than by changes in the expression of their genes. PMID- 8412695 TI - The absence of a surface protease, OmpT, determines the intercellular spreading ability of Shigella: the relationship between the ompT and kcpA loci. AB - A large plasmid-encoded protein, VirG, on the bacterial surface is essential for the spreading of Shigella by eliciting polar deposition of filamentous actin in the cytoplasm of epithelial cells. VirG expression from the large plasmid is diminished greatly when it is introduced into Escherichia coli K-12 from Shigella. In an attempt to identify factors affecting VirG expression, we found that the absence of the ompT gene, encoding outer membrane protease OmpT, restored full production of VirG protein to E. coli K-12. Conversely, upon introduction of the ompT gene of E. coli K-12 into Shigella, spreading ability was completely abolished, probably because of the proteolytic degradation of VirG protein by OmpT. Analysis of the DNA sequence of the ompT region indicated that the absence of the ompT gene occurred in Shigella and enteroinvasive E. coli strains, and that the absent DNA segment corresponded to a remnant lambdoid phage structure found in E. coli K-12, which encompasses a 21 kb DNA segment spanning from argU through to the ompT genes. Since ompT is located near purE in E. coli K 12 and a virulence locus for provoking keratoconjunctivitis in the eyes of guinea pigs, named kcpA, is located near purE in S. flexneri, and the two loci are involved in VirG expression, the KcpA- mutants of S. flexneri 2a constructed were examined for correlation between acquisition of ompT and VirG degradation. Our data suggest that the previous recognition of a kcpA locus in S. flexneri is the result of transfer of the ompT gene from E. coli K-12, giving rise to a KcpA- phenotype. These results indicate that the lack of OmpT protease confers upon Shigella the ability to spread into adjacent epithelial cells. PMID- 8412696 TI - Orientation of porin channels in the outer membrane of Bordetella pertussis. AB - We have examined the surface topography and channel connectivity of a naturally crystalline porin that is known to be functional, and whose structure has not been perturbed by detergent extraction. A three-dimensional density map, calculated from two independent tilt series of negatively stained cell envelopes, reveals three separate channels per trimer on one side (the 'smooth' side), and a single common opening at the other ('rough') side. This arrangement is consistent with the molecular structures recently determined at high resolution by X-ray crystallography for three other porins after detergent solubilization, and implies that the Bordetella pertussis porin may have the same kind of folding. Surface relief maps calculated from electron micrographs of cell envelopes contrasted by unidirectional shadowing clearly show that the side with single opening (i.e. the rough side) represents the external surface. PMID- 8412697 TI - Erwinia stewartii WtsA, a positive regulator of pathogenicity gene expression, is similar to Pseudomonas syringae pv. phaseolicola HrpS. AB - Erwinia stewartii contains a large cluster of wts genes that are required by this bacterium for pathogenicity on corn plants. Three complementation groups within the right half of this cluster, wtsA, wtsC, and wtsB, were previously identified. In this study, WtsA was found to be a positive activator of wtsB::lacZ expression. The wtsA locus was sequenced and a single open reading frame is present within the wtsA locus, which has the capacity to encode a 323 amino acid polypeptide. A corresponding 38 kDa protein was observed in Escherichia coli minicells containing the cloned wtsA gene. The predicted WtsA polypeptide has significant similarity to HrpS from Pseudomonas syringae pv. phaseolicola, as well as other members of the NtrC class of prokaryotic regulatory proteins. Similar to other genes activated by NtrC regulators, wtsB::lacZ expression in E. coli was dependent upon rpoN. PMID- 8412698 TI - Differentiation of Pseudomonas aeruginosa into the alginate-producing form: inactivation of mucB causes conversion to mucoidy. AB - Mucoidy in Pseudomonas aeruginosa is a critical virulence factor associated with chronic respiratory infections in cystic fibrosis. A cluster of three tightly linked genes, algU, mucA and mucB located at 67.5 min, controls development of mucoid phenotype. This locus is allelic with a group of mutations (muc) associated with conversion into constitutively mucoid forms. One of the genes previously characterized in this region, algU, is absolutely required for the transcriptional activation of algD, a critical event in the establishment of mucoidy. AlgU is homologous to the alternative sigma factor sigma H (Spo0H) controlling sporulation and competence in Bacillus. Two genes downstream of algU, mucA and mucB were further characterized in this study. Previous complementation studies have demonstrated that mucA is required for suppression of mucoidy in the muc-2 strain PAO568. In this work, complementation analysis indicated that, in addition, mucB was required for suppression of mucoidy in the muc-25 strain PAO581, and for enhanced complementation of the muc-2 mutation in PAO568. The complete nucleotide sequence of mucA and mucB was determined. Insertional inactivation of mucB on the chromosome of the standard genetic strain PAO resulted in mucoid phenotype, and in a strong transcriptional activation of algD. Thus, a loss of mucB function is sufficient to cause conversion of P. aeruginosa into the mucoid phenotype. Since the algU-mucA-mucB region is a general site where muc mutations have been mapped, it is likely that mucB participates in the emergence of mucoid forms. Both mucA and mucB play a regulatory role in concert with the sigma-like factor AlgU; all three genes, along with signal transduction and histone-like elements, control differentiation of P. aeruginosa into the mucoid phenotype. PMID- 8412699 TI - Inhibition of translational initiation in Saccharomyces cerevisiae by secondary structure: the roles of the stability and position of stem-loops in the mRNA leader. AB - A new modular gene-expression system for use in studies of translational control in Saccharomyces cerevisiae was constructed. A GAL::PGK fusion promoter (GPF) directed the inducible synthesis of mRNAs initiated at a single major site. A series of leader sequences were tested in combination with each of two reporter genes (encoding chloramphenicol acetyl transferase (cat) and luciferase (luc)). Stem-loop structures of three different sizes and predicted stabilities were inserted into each of two different unique restriction sites in the leader. After correction for relative mRNA abundance, a stem-loop of predicted stability equivalent to approximately -18 kcal mol-1 inhibited translation by up to 89%. The degree of inhibition exerted by the other stem-loops correlated positively with their predicted stabilities. Combinations of two stem-loops at different sites yielded an inhibitory effect greater than that of either individual stem loop alone. Similar inhibitory effects were observed with both reporter genes. However, inhibition of translation, particularly of the cat gene, was more effective when the stem-loop was positioned close to the start codon rather than at the 5' end of the leader. The observed results reflect an important form of post-transcriptional control that is expected to act on a large number of genes in yeast. PMID- 8412700 TI - NAD+ binding to the Escherichia coli K(+)-uptake protein TrkA and sequence similarity between TrkA and domains of a family of dehydrogenases suggest a role for NAD+ in bacterial transport. AB - The nucleotide sequence of trkA, a gene encoding a surface component of the constitutive K(+)-uptake systems TrkG and TrkH from Escherichia coli, was determined. The structure of the TrkA protein deduced from the nucleotide sequence accords with the view that TrkA is peripherally bound to the inner side of the cytoplasmic membrane. Analysis by a dot matrix revealed that TrkA is composed of similar halves. The N-terminal part of each TrkA half (residues 1-130 and 234-355, respectively) is similar to the complete NAD(+)-binding domain of NAD(+)-dependent dehydrogenases. The C-terminal part of each TrkA half (residues 131-233 and 357-458, respectively) aligns with the first 100 residues of the catalytic domain of glyceraldehyde-3-phosphate dehydrogenase. Strong u.v. illumination at 252 nm led to cross-linking of NAD+ or NADH, but not of ATP to the isolated TrkA protein. PMID- 8412701 TI - Identification of a new nuclear gene (CEM1) encoding a protein homologous to a beta-keto-acyl synthase which is essential for mitochondrial respiration in Saccharomyces cerevisiae. AB - We have analysed a new gene, CEM1, from Saccharomyces cerevisiae. Inactivation of this gene leads to a respiratory-deficient phenotype. The deduced protein sequence shows strong similarities with beta-keto-acyl synthases or condensing enzymes. Typically, enzymes of this class are involved in the synthesis of fatty acids or similar molecules. An analysis of the mitochondrial lipids and fatty acids shows no major difference between the wild type and deleted strains, implying that the CEM1 gene product is not involved in the synthesis of the bulk fatty acids. Thus it is possible that the CEM1 protein is involved in the synthesis of a specialized molecule, probably related to a fatty acid, which is essential for mitochondrial respiration. PMID- 8412702 TI - Escherichia coli endoribonuclease RNase E: autoregulation of expression and site specific cleavage of mRNA. AB - Mutations in the Escherichia coli rne (ams) gene have a general effect on the rate of mRNA decay in vivo. Using antibodies we have shown that the product of the rne gene is a polypeptide of relative mobility 180 kDa. However, proteolytic fragments as small as 70 kDa, which can arise during purification, also exhibit RNase E activity. In vitro studies demonstrate that the rne gene product, RNase E, is an endoribonuclease that cleaves mRNA at specific sites. RNase E cleaves rne mRNA and autoregulates the expression of the rne gene. In addition we demonstrate RNase E-dependent endonucleolytic cleavage of ompA mRNA, at a site known to be rate-determining for degradation and reported to be cleaved by RNase K. Our data are consistent with RNase K being a proteolytic fragment of RNase E. PMID- 8412703 TI - The ntrBC genes of Rhizobium leguminosarum are part of a complex operon subject to negative regulation. AB - We report here that ntrB and ntrC genes of Rhizobium leguminosarum biovar phaseoli are cotranscribed with an open reading frame (called ORF1) of unknown function. The promoter region of the ORF1-ntrB-ntrC operon was mapped immediately upstream of ORF1 and two in vivo transcription initiation sites were identified, both preceded by -35/-10 promoter consensus sequences. Some major aspects differentiate R. leguminosarum from the enteric nitrogen regulatory system: the ntrBC genes are cotranscribed with ORF1 which is homologous to an ORF located upstream of ntrBC of R. capsulatus and to the ORF1 located upstream of the fis gene of Escherichia coli; ntrBC are not transcribed from a -24/-12 promoter and are only autogenously repressed. Moreover, the intracellular concentration of the NtrC protein increases when the bacterium is grown on ammonium salts, while under the same conditions the promoter of one of its target genes, glnII, is 12 times less active. PMID- 8412704 TI - An accessory gene, lipB, required for the production of active Pseudomonas glumae lipase. AB - Pseudomonas glumae PG1 is able to secrete lipase into the extracellular medium. The lipase is produced as a precursor protein, with an N-terminal signal sequence. A second open reading frame (ORF) was found immediately downstream of the lipase structural gene, lipA, a situation found for the lipases of some other Pseudomonas species. Inactivation of this ORF resulted in a lipase-negative phenotype, indicating its importance in the production of active extracellular lipase. The ORF, lipB, potentially encodes a protein of 353-amino-acid residues, having a hydrophobic N-terminal (amino acids 1 to 90) and a hydrophilic C terminal part. As a first step in determining the role of LipB, its subcellular location was determined. The protein was found to fractionate with the inner membranes. The expression of fusions of lipB fragments with phoA revealed an N(in)-C(out) topology for the LipB protein, which was confirmed by protease accessibility studies on EDTA-permeabilized cells and on inverted inner membrane vesicles. These and other results indicate that most of the LipB polypeptide is located in the periplasm and anchored to the inner membrane by an N-terminal transmembrane helix, located between amino acids 19 and 40. PMID- 8412705 TI - Role of the lipB gene product in the folding of the secreted lipase of Pseudomonas glumae. AB - The LipB protein of Pseudomonas glumae is essential for the production of active extracellular lipase encoded by the lipA gene. When lipase is overproduced in P. glumae in the absence of a functional lipB gene, the enzyme accumulates intracellularly in an inactive conformation. Heterologous expression of the lipase in Pseudomonas aeruginosa, Bacillus subtilis and Escherichia coli indicated that LipB is not directly involved in the translocation of the lipase across the inner or outer membrane. However, the presence of LipB was essential for obtaining active lipase and had a profound influence on the stability of the protein to proteolytic degradation. Inactive lipase, produced in the absence of LipB could be activated in vitro by unfolding and refolding, which demonstrates that LipB activity is not responsible for an essential covalent modification of the enzyme. We propose that LipB is a lipase-specific foldase. Furthermore, proper folding of the lipase in the periplasm appears to be essential for Xcp mediated translocation across the outer membrane. PMID- 8412706 TI - Chemotaxis plays a role in the social behaviour of Myxococcus xanthus. AB - Myxococcus xanthus is a Gram-negative bacterium that glides on a solid surface and displays a wide range of social behaviour including microbial development. The frz genes are homologues to the chemotaxis genes of Escherichia coli and Salmonella typhimurium and have been shown to be involved in microbial development. However, chemotaxis has never been clearly demonstrated in Myxococcus. In this study, we showed that M. xanthus exhibited tactic movements to many chemicals when they were subjected to steep and stable chemical gradients. M. xanthus was observed to spread into areas with abundant nutrients like yeast extract or Casitone and avoid areas with no nutrients or repellents (short-chain alcohols or DMSO). Responses to attractants and repellents were additive. Movement towards attractants or away from repellents required the frz genes and was correlated with methylation or demethylation of FrzCD, a methyl accepting taxis protein. Furthermore, the frz genes were found to be required for both fruiting body formation during starvation and swarming in nutrient-rich medium. In wild-type strains, cells near the colony edge were observed to swarm towards the surrounding growth medium and to contain highly methylated FrzCD; cells near the colony centre contained mainly demethylated FrzCD and showed directed movement towards the colony edge. FrzCD was also found to be methylated during the aggregation stage of fruiting body formation on agar but largely demethylated in cells shaken in liquid starvation media. An frzE mutant failed to exhibit directed cell movements and no longer showed modification of FrzCD under these conditions. These observations suggest that M. xanthus does show chemotactic movements, that these movements require the frz genes, and that chemotaxis plays a very important role in the social behaviour of this organism. PMID- 8412707 TI - Transcription at different salinities of Haloferax mediterranei sequences adjacent to partially modified PstI sites. AB - Two genomic sequences from the halophilic archaeon Haloferax mediterranei, where we had found PstI restriction-pattern modifications depending on the salinity of the growth medium, have been studied. A markedly salt-dependent differential expression has been detected in the nearby regions. Two of the open reading frames characterized correspond to two of the differentially expressed transcripts. In both cases the PstI sites were included in purine-pyrimidine alternancies suggestive of Z-DNA structures and located in non-coding regions with frequent repetitive motifs. A long alternating adenine-thymine tract also appears in the upstream regions of one of these open reading frames. A possible role of local DNA configuration in osmoregulation in this organism is discussed. PMID- 8412708 TI - Evolution of penicillin resistance in Streptococcus pneumoniae; the role of Streptococcus mitis in the formation of a low affinity PBP2B in S. pneumoniae. AB - Penicillin-resistant strains of Streptococcus pneumoniae possess forms of penicillin-binding proteins (PBPs) that have a low affinity for penicillin compared to those from penicillin-sensitive strains. PBP genes from penicillin resistant isolates are very variable and have a mosaic structure composed of blocks of nucleotides that are similar to those found in PBP genes from penicillin-sensitive isolates and blocks that differ by up to 21%. These chromosomally encoded mosaic genes have presumably arisen following transformation and homologous recombination with PBP genes from a number of closely related species. This study shows that PBP2B genes from many penicillin resistant isolates of S. pneumoniae contain blocks of nucleotides originating from Streptococcus mitis. In several instances it would appear that this material alone is sufficient to produce a low affinity PBP2B. In other examples PBP2B genes possess blocks of nucleotides from S. mitis and at least one additional unidentified species. Mosaic structure was also found in the PBP2B genes of penicillin-sensitive isolates of S. mitis or S. pneumoniae. These mosaics did not confer penicillin resistance but nevertheless reveal something of the extent to which localized recombination occurs in these naturally transformable streptococci. PMID- 8412709 TI - Judgmental overshadowing: further evidence of cue interaction in contingency judgment. AB - We investigated a phenomenon called judgmental overshadowing. Subjects predicted whether each of several patients had a disease on the basis of whether or not the patient had each of two symptoms. For all the subjects, the presence of the disease was moderately contingent on the presence of one of the symptoms (S1). In Condition 1 of our first experiment, the presence of the disease was highly contingent on the presence of the other symptom (S2). In Condition 2, the presence of the disease was independent of S2. Judgmental overshadowing occurred in that the S1-disease contingency was judged to be stronger in Condition 2 than in Condition 1. Subsequent experiments showed that judgmental overshadowing depends little on the form of the judgment, is not due to a response bias or contrast effect, and does not depend on subjects' actively diagnosing each patient. These results are consistent with, and are generally predicted by, an associative-learning model of contingency judgment. PMID- 8412710 TI - Induction of combination rules in two-dimensional function learning. AB - Previous studies have typically found that when people learn to combine two dimensions of a stimulus to select a response, they learn additive combination rules more easily than nonadditive (e.g., multiplicative) ones. The present experiments demonstrate that in some situations people can learn multiplicative rules more easily than other (e.g., additive) rules. Subjects learned to produce specified response durations when presented with stimulus lines varying in length and angle of orientation. When stimuli and correct responses were related by a multiplicative combination of power functions, learning was relatively easy (Experiment 1). In contrast, systematic response biases occurred during the early phases of learning an additive combination of linear functions (Experiment 2) and a more complex (nonadditive and nonmultiplicative) combination of linear functions (Experiment 3), suggesting that people have a tendency to induce a multiplicative combination of power functions. However, the initial biases decreased with practice. These results are explained in terms of a revised adaptive regression model of function learning originally proposed by Koh and Meyer (1991). Differences between the present results and previous results in the literature are discussed. PMID- 8412711 TI - Memory for mental models of spatial descriptions: an episodic-construction-trace hypothesis. AB - Three experiments, based on Mani and Johnson-Laird's (1982) study, tested memory for spatial descriptions. In Phase 1 of each experiment, subjects judged whether diagrams matched verbal descriptions that were either determinate or indeterminate. In Phase 2, subjects attempted to recognize the descriptions studied in Phase 1. Mani and Johnson-Laird reported a crossover: Gist memory was better for determinate descriptions and verbatim memory was better for indeterminate descriptions. This crossover suggests that mental models are remembered. All three new experiments failed to replicate the crossover, challenging whether models are preserved in episodic memory. It was hypothesized instead that episodic memory records the mental operations by which models are constructed. This hypothesis accounted for the findings of all three experiments. In Experiments 1 and 2, performance varied with the construction-trace overlap between recognition test descriptions. In Experiment 3, performance was depressed if the order of sentences within descriptions was altered between study and test. PMID- 8412712 TI - Orientation-invariant transfer of training in the identification of rotated natural objects. AB - The effects of stimulus orientation on naming were examined in two experiments in which subjects identified line drawings of natural objects following practice with the objects at the same or different orientations. Half the rotated objects were viewed in the orientation that matched the earlier presentations, and half were viewed at an orientation that mismatched the earlier presentations. Systematic effects of orientation on naming time were found during the early presentations. These effects were reduced during later presentations, and the size of this reduction did not depend on the orientation in which the object had been seen originally. The results are consistent with a dual-systems model of object identification in which initially large effects of disorientation are the result of a normalization process such as mental rotation, and in which attenuation of the effects is due to a shift from the normalization system to a feature/part-based system. PMID- 8412713 TI - The loss of repetition priming and automaticity over time as a function of degree of initial learning. AB - Two experiments were performed to investigate the buildup of repetition priming in a lexical decision task with repeated presentations and its decline over the course of 2 months. Priming was found to accumulate as a power function of presentations and to decline as a power function of time. Accuracy measures indicated that the loss rate of priming was unaffected by the amount of initial priming. Response time measures indicated the same result when the experiments were analyzed separately; however, when the data were combined, increased initial priming was associated with greater losses in priming over time. The data were interpreted in terms of automaticity, and the power function decline in priming was taken as support for memory-based models of automaticity. Possible ways to incorporate forgetting into memory-based theories of automaticity are discussed. PMID- 8412714 TI - Context effects in repetition priming are sense effects. AB - This article reports three experiments that investigate the role of context in repetition priming using a lexical decision task. The experiments show that repetition priming is either eliminated or significantly reduced if a change in context also alters the perceived sense of a nonhomographic target word. If perceived sense is not altered, a change in context is inconsequential. This points to the important role played by perceived sense in repetition priming. An explanation within a sense-specific activation framework is proposed in preference to a modified processing view. PMID- 8412715 TI - Selective attention to Stroop dimensions: effects of baseline discriminability, response mode, and practice. AB - The framework of dimensional interaction was used to test the hypothesis that the Stroop effect is partially rooted in mismatches in baseline discriminability, with stimulus differences along the word dimension typically exceeding stimulus differences along the color dimension. Subjects made speeded classifications, with either keypresses or vocalizations, of either words or colors. Stroop congruity and Garner interference were measured under conditions in which discriminabilities were (1) matched (Experiments 1 and 4), (2) mismatched in favor of colors (Experiment 2), or (3) mismatched in favor of words (Experiment 3). When matched, colors and words appeared separable, with small interactive effects being reduced or eliminated through practice. When mismatched, asymmetric Stroop and Garner effects emerged, with the more discriminable dimension disrupting classification of the less discriminable dimension. Asymmetric effects were obtained in both response modes, and were not alleviated by practice. We conclude that (1) the Stroop effect is an optional effect, and (2) unequal discriminability causes a mandatory failure of selective attention. PMID- 8412716 TI - Expected endings and judged duration. AB - In four experiments, the predictions of an expectancy/contrast model (Jones & Boltz, 1989) for judged duration were evaluated. In Experiments 1 and 2, listeners estimated the relative durations of auditory pattern pairs that varied in contextual phrasing and temporal contrast. The results showed that when the second pattern of a pair either seems to (Experiments 1 and 2) or actually does (Experiment 2) end earlier (later) than the first, subjects judge it as being relatively shorter (longer). In Experiment 3, listeners heard single patterns in which notes immediately preceding the final one were omitted. Timing of the final (target) tone was varied such that it was one beat early, on time, or one beat late. Listeners' ratings of target tones revealed systematic effects of phrasing and target timing. In Experiment 4, listeners temporally completed (extrapolated) sequences of Experiment 3 that were modified to exclude the target tone. The results again showed that phrase context systematically influenced expectancies about "when" sequences should end. As a set, these studies demonstrate the effects of event structure and anticipatory attending upon experienced duration and are discussed in terms of the expectancy/contrast model. PMID- 8412717 TI - Recall and articulation of English and Chinese words by Chinese-English bilinguals. AB - Three groups of subjects were tested to investigate the effect of language on the relationship between recall span and articulation rate. Native English-speaking monolinguals and native Chinese-speaking monolinguals recalled only English or Chinese words, respectively. Chinese-English bilinguals recalled both English and Chinese words. Articulation rates for English and Chinese monolinguals and Chinese-English bilinguals in each language were also obtained. When recall span was regressed on articulation rate, the slopes for Chinese and English words were significantly different for the Chinese-English bilinguals. This difference was not due to language proficiency but to phonological differences between English and Chinese. PMID- 8412718 TI - Is there really very rapid forgetting from primary memory? The role of expectancy and item importance in short-term recall. AB - In two experiments, subjects recalled one of two letter segments following a digit-filled retention interval. In Experiment 1, recall expectancy was manipulated by using precues that correctly informed or misinformed subjects concerning which letter segment would be tested for recall. In Experiment 2, item importance was varied by precuing one segment as important but requiring that the uncued segment be recalled first. Recall performance was very low under conditions of low expectancy and low segment importance, but the slopes of the retention functions did not demonstrate more rapid forgetting than under standard conditions. The previous observations of very rapid forgetting from primary memory may be a function of an elevated initial recall level in the earlier studies. Our retention functions were compared with predictions of the Estes perturbation model. The findings suggested that when secondary memory processes were reduced, forgetting order information from primary memory occurred at the same rate as that estimated on the basis of previous studies using the standard distractor task. PMID- 8412719 TI - Distinctiveness and serial position effects in recognition. AB - Digitized photographs of snowflakes were presented for a recognition test after retention intervals of varying durations. While overall accuracy and discrimination remained constant, as the retention interval increased, primacy increased from chance to reliably better than chance while recency decreased to chance levels. A variation of Murdock's (1960) distinctiveness model accounted for the changing primacy and recency effects observed in both between- and within subjects designs. The generality of the model was examined in two different paradigms: lexical access during sentence processing, and free recall in the continual distractor paradigm. In both cases, the model made accurate qualitative predictions for both latency and accuracy measures. PMID- 8412720 TI - Decomposing adult age differences in symbol arithmetic. AB - A componential analysis was conducted to determine the locus of adult age differences in symbol arithmetic. Measures of the duration of two proposed components, substitution of digits for symbols and the addition or subtraction of the digits resulting from these substitutions, were obtained from 52 young adults and 52 older adults. Tests of working memory, perceptual speed, motor speed, and associative learning were also administered to all subjects. The results were most consistent with an interpretation postulating that the speed of many different cognitive processes decreases with increased age. Considerable age related variance remained in the measures of symbol arithmetic performance after statistical control of working memory and associative learning performance, casting doubt on alternative hypotheses of the source of age-related differences in this task. PMID- 8412721 TI - Conceptual metaphors are not automatically accessed during idiom comprehension. AB - Do conceptual analogies motivate idiom use and comprehension in discourse? For example, a story in which a person is described as fuming would be analogically consistent with an idiom such as blew her top, but inconsistent with an idiom such as bite his head off. Earlier work by Nayak and Gibbs (1990) had suggested that people use such analogical information during idiom comprehension. We replicated their findings in an idiom choice task, suggesting that people can indeed make use of such knowledge. However, when reading times were used to assess idiom comprehensibility, no effects of analogical consistency were found. We conclude that conceptual analogies play little, if any, role in idiom comprehension unless people have the time (and motivation) to make considered judgments. PMID- 8412722 TI - Funding, expenditures, and staffing of mental health services in state adult correctional facilities: United States, 1988. AB - State-by-State data concerning the sources of funding, expenditures, and staffing for mental health services in State correctional facilities are reported for 1988 from the first national survey of prison mental health services conducted by the Center for Mental Health Services. Among all States, the total annual funding per prison inmate for mental health services varied widely, ranging from $5.67 to $3,159.41 per inmate, with a mean of $469.67 and median of $303.48 per inmate. States that administered 24-hour hospital mental health care to prisoners solely through the Department of Corrections (DOC) also tended to fund all types of mental health services solely through the DOC. However, in States where the Department of Mental Health (DMH) had primary administrative responsibility for 24-hour hospital mental health care, funding sources for all types of mental health services available to prison inmates were more likely to be mixed--i.e., funded through DOC as well as through DMH and other administrative entities. Master's-level mental health providers outnumbered doctoral-level professionals by more than two to one. At both of these educational levels, psychologists were numerically the largest category of provider, followed by social workers among master's-trained professionals, and followed by psychiatrists among those with doctorates. The single largest category was mental health workers with bachelor's level training or less; this group accounted for about 44 percent of all mental health staff on State prison payrolls. PMID- 8412723 TI - Demographic and diagnostic characteristics of inmates receiving mental health services in state adult correctional facilities: United States, 1988. AB - The demographic and diagnostic characteristics of inmates in State adult correctional facilities who received 24-hour hospital mental health care, residential treatment care, and counseling/therapy in 1988 are reported by State and by type of administrative auspices under which the services are provided. Rates under treatment for 24-hour hospital mental health care were highest for the youngest (under 18) and oldest (65 and over) age groups, for females, and for whites. For counseling/therapy, rates were also highest for the youngest, for females, and for whites, but they declined with age. Rates in residential treatment were highest for the young and old and for whites, but about equal for males and females. Primary diagnoses of major psychoses predominated in 24-hour hospital mental health care. In residential treatment, a comparatively small proportion of the caseload had major psychotic disorders and a comparatively large proportion had substance abuse and mental retardation diagnoses. In counseling/therapy, personality disorders predominated. Individual State figures vary widely on these characteristics, both within and between service auspice types. PMID- 8412724 TI - The "lethal chamber": further evidence for the euthanasia option. AB - Historical discussions of the euthanasia or "lethal chamber" option in relation to people with mental retardation were presented. Positions were classified into those (a) dismissing the option, (b) presenting ambivalent views, and (c) showing outright support. Tredgold's forthright views in support of euthanasia for "idiots" and "imbeciles" were highlighted. Eugenic beliefs in the primacy of heredity over environment in determining mortality and the positive role of natural selection in race betterment were presented. I proposed that these beliefs were used to condone, if not encourage, the poor conditions characteristic of large, segregated institutions during the early decades of this century and their high rates of mortality from infectious diseases such as tuberculosis. The conclusion was drawn that euthanasia may thus have been implemented, albeit unofficially, in at least some institutions. PMID- 8412725 TI - Institutionalization, involuntary sterilization, and mental retardation: profiles from the history of the practice. AB - For much of the 20th century, the practice of sterilization was a common accompaniment to institutionalization for individuals with mental retardation. Following the Supreme Court's decision in Buck v. Bell (1927) supporting the practice, numerous states passed legislation and, consequently, over 60,000 individuals with mental retardation were sterilized in this country. In the present study we analyzed data on 212 individuals in Virginia who were sterilized. Our focus included gender, age at and date of sterilization, level of mental retardation, and location of subsequent discharge. The nature of these findings within the general context of the practice of sterilization was discussed. PMID- 8412726 TI - Reducing barriers to sex education for adults with mental retardation. AB - Techniques were identified that staff of agencies serving adults or adolescents with mental retardation perceived would enable them to provide systematic sex education and counseling. Through the use of a structured interview process, we obtained opinions of professionals from 16 agencies. On the basis of these interviews and a thorough review of related literature, we made recommendations aimed at increasing professionals' ability to provide systematic sex education and counseling for individuals with mental retardation. Suggestions for further study were presented. PMID- 8412727 TI - Awareness and knowledge of fragile X syndrome among special educators. AB - A questionnaire to assess awareness and knowledge of various syndromes was mailed to a random sample of elementary special education teachers in Colorado. Approximately half of the questionnaires were completed and returned. Results indicate that significantly more respondents had knowledge of Down syndrome than of fetal alcohol or fragile X syndromes. Respondents' level of education, date of most recent degree, and years of experience were not associated with greater awareness nor knowledge of fragile X syndrome. Respondents expressed interest in a broad array of fragile X in-service training topics. PMID- 8412728 TI - Drama: a medium to enhance social interaction between students with and without mental retardation. AB - Twenty-four fifth grade (or equivalent age) students were assigned to one of two treatment conditions consisting of equal numbers of students from regular and special education classes. One group was involved in a dramatic games condition culminating in a theatrical performance planned by the group. The second group was involved in a noncompetitive games condition culminating in a demonstration of games developed by the group. Observational data recording initiations and receptions of prosocial bids were collected for all subjects receiving special education services within both groups. A sociometric measure was used to assess the perceived quality of friendships both before and after the interventions. Results showed that students with mental retardation in the drama condition were targeted for positive social interactions by peers without mental retardation significantly more and were more highly regarded as friends than were those in the games condition. PMID- 8412729 TI - Staff stressors at a developmental center and state hospital. AB - Clinicians and administrators (N = 322) at a large state hospital/developmental center were surveyed regarding stressful experiences at work. A factor analysis of stress reports yielded two underlying constructs: Lack of Administrative Control and Practice-Related Stress. A subsequent regression analysis found that philosophical opposition to behavior therapy significantly predicted job stress. No differences were found between stress levels of staff members who worked with individuals who had mental retardation versus mental illness. Differences existed in reported stress by job categories; nurses, developmental specialists, and psychiatric technicians reported greater job-related stress than did other professional groups. Understanding the nature of job-related stress is necessary for developing stress management programs for in-patient personnel. PMID- 8412730 TI - Managing the care of patients with mental retardation: a survey of physicians. AB - A questionnaire regarding the clinical management of patients with mental retardation living in the community was mailed to a 50% random sample of physicians practicing in Maine; the response rate was 56%. Almost 80% of respondents reported that they were currently providing care to patients with mental retardation. Survey results identified the major issues confronting physicians treating such patients and quantitatively assessed their importance from the physicians' perspective. Inadequate medical information was identified as the greatest obstacle to patient care. Physicians' ratings of the utility of specific measures designed to lessen these obstacles were presented. Generally, continuing education was viewed as less useful than practice support measures. PMID- 8412731 TI - Acceptability of behavioral treatments: influence of knowledge of behavioral principles. AB - The relation of staff knowledge of behavioral principles to ratings of treatment acceptability for interventions used in the management of self-injurious behavior (SIB) was examined with 57 direct-care staff members from an ICF/MR. Case descriptions of an adult with severe mental retardation who exhibited SIB were presented, followed by descriptions of six behavioral interventions rated on acceptability. Subjects also completed a 25-item measure of general knowledge of behavioral principles. Results indicated that staff age and educational attainment but not treatment acceptability ratings were significantly related to knowledge scores. Implications of the findings were discussed. PMID- 8412732 TI - Collaborative efforts: developing clinical experiences for nursing students in public sector settings. AB - In the present paper we described the Nurse Academic Collaboration Task Force, which was formed in 1989 by the Texas Mental Health and Mental Retardation (TXMHMR) Commissioner in order to facilitate linkages between academia and state facilities. These efforts have led to increased academic collaboration and linkage with facilities. The task force has identified areas of practice opportunities for all levels of student education programs as well as barriers to collaboration. Collaborative strategies may also lead to better nurse recruitment efforts by TXMHMR facilities. Finally, these educational experiences should prepare nurses in varied settings to provide care for individuals with mental retardation. PMID- 8412733 TI - Teacher report versus Adaptive Behavior Scale in assessment of mental retardation. PMID- 8412734 TI - Glucose enters mitochondrial metabolism via both carboxylation and decarboxylation of pyruvate in pancreatic islets. AB - The routes by which glucose-derived pyruvate is metabolized to enter mitochondrial pathways to stimulate insulin release in the pancreatic beta cell are unknown. The 14CO2 ratios assay was used to estimate the fractions of glucose derived pyruvate that enter the citric acid cycle by carboxylation and decarboxylation in pancreatic islets. Pyruvate 14CO2 ratios were estimated with glucose, and acetate 14CO2 ratios were estimated with succinate methyl ester. Glucose and methyl succinate, which are insulin secretagogues, gave more accurate ratios than ratios estimated with pyruvate and acetate, which do not initiate insulin release. The results indicate that relatively equal amounts of pyruvate enter the citric acid cycle by carboxylation and decarboxylation. PMID- 8412735 TI - Serum insulin distributions and reproducibility of the relationship between 2 hour insulin and plasma glucose levels in Asian Indian, Creole, and Chinese Mauritians. Mauritius NCD Study Group. AB - The relationship of 2-hour (post-75 g oral glucose) serum insulin levels with plasma glucose levels was studied in a population-based random sample comprising 2,627 Hindu Indians, 685 Muslim Indians, 1,351 Creoles (African, European, and Indian admixture), and 415 Chinese from the Indian Ocean island of Mauritius. Known diabetic subjects taking oral hypoglycemic drugs or insulin were excluded from these analyses; 64% of all diabetic subjects had usable glucose and insulin data. Both fasting and 2-hour postload insulin levels were significantly higher in women than in men, and levels in both sexes were significantly greater in Hindu and Muslim Indian subjects than in Creoles or Chinese even after controlling for differences in age, body mass index (BMI), waist to hip ratio (WHR), and plasma glucose level. Levels in Muslims were higher than those in Hindus; it was unclear whether these ethnic differences represented hereditary or unmeasured environmental factors closely associated with ethnicity. All four ethnic groups demonstrated similar inverted U- or V-shaped curves when 2-hour insulin was plotted against either basal or 2-hour glucose. Both quadratic (U) and two-piece (V) regression models improved over linear models for 2-hour insulin versus either fasting or 2-hour glucose in all ethnic groups, although in statistical terms they were good models only for the 2-hour glucose comparison. The two-piece models were associated with modest increases in R2 compared with the quadratic models, but it was not possible to precisely determine optimal turning points with either model. However, in all ethnic groups, 2-hour insulin levels decreased above glucose levels of 7.1 to 7.8 (fasting) and 11.3 to 13.5 mmol/L (2-hour) in quadratic models, and 7.5 to 9.5 (fasting) and 8.5 to 10.5 mmol/L (2-hour) in two-piece models. The shape and point of inflection of the quadratic and two-piece curves were influenced little by gender, obesity, fat distribution, and physical activity. These results are in accord with those observed in cross-sectional and longitudinal studies in other ethnic groups, and support the generality of the plasma glucose levels currently used to define diabetes mellitus, which physiologically correspond with a decrease in beta-cell responsiveness to glucose. Asian Indians appear to have an ethnic propensity to hyperinsulinemia that is not explained by obesity or adverse fat distribution. PMID- 8412736 TI - Effects of ethanol on urinary acidification and on gluconeogenesis by isolated renal tubules. AB - Class I alcohol dehydrogenase (ADH) is present in the kidney of rats. Rats fed an alcohol-containing diet long-term had higher urinary pH and reduced titratable acidity compared with pair-fed controls; rates of ammonium excretion were unchanged. The effects of ethanol on the metabolism of isolated renal tubules were then studied. Gluconeogenesis from lactate, pyruvate, or glutamine was not inhibited by 10 mmol/L ethanol during 30- or 60-minute incubations, although there was a trend toward increased lactate/pyruvate ratios at 30 minutes in the presence of ethanol. When the medium was also supplemented with oleate, glucose synthesis from most substrates was decreased, and the addition of ethanol inhibited glucose synthesis dramatically. This interaction between oleate and ethanol was not abolished by 4-methylpyrazole, an inhibitor of ADH. This effect of ethanol was highly dependent on the concentration of oleate present in the medium and was not observed with palmitate or decanoate; the inhibition was reversed by increasing the medium concentration of albumin. We conclude that ethanol may mildly perturb the redox state of isolated kidney tubules without inhibiting glucose synthesis, and that ethanol and oleate interact to inhibit renal gluconeogenesis by a mechanism highly dependent on the fatty acid concentration. The mechanism by which ethanol in the diet reduces renal acid excretion remains unknown. PMID- 8412737 TI - Protein and glucose fuel kinetics and hormonal changes in elderly trauma patients. AB - We investigated the responsiveness of whole-body protein and glucose kinetics to severe trauma and the role of regulatory hormones in the early catabolic "flow phase" of injury in a group of patients ranging in age from 19 to 85. Energy metabolism (indirect calorimetry), protein kinetics (primed-constant infusion of [15N]glycine), glucose metabolism (primed-constant infusion of [U-14C]glucose and [6-3H]glucose), and circulating hormone levels were measured during basal conditions within 48 to 60 hours after injury when the patients were receiving maintenance fluids without N or calories. Experimental data were analyzed as two groups (young, age 18 to 59, and elderly, age 60 to 85) and also as linear function of advancing age. The geriatric trauma (GT) group lost less N (11.9 +/- 1.3 v 17.7 +/- 1.7 g N/d, P = .025) than the younger group, mainly due to a significantly decreased whole-body protein breakdown (WBPB) rate. Despite a similar production and oxidation rate of glucose, hyperglycemia was more exaggerated in the elderly group. Advancing age resulted in significant positive correlations with plasma glucose levels and negative correlations with circulating growth hormone levels, urinary nitrogen loss, and protein turnover. These results suggest an age- and body composition-related reduced energy and protein metabolic response to severe trauma in elderly individuals. PMID- 8412738 TI - Hepatic conversion of amino nitrogen to urea nitrogen in hypothyroid patients and upon L-thyroxine therapy. AB - Conflicting studies have been reported regarding the influence of thyroid hormones on hepatic nitrogen metabolism and liver metabolic activity. We studied urea N synthesis rate (UNSR), functional hepatic N clearance (FHNC), galactose elimination capacity, and antipyrine clearance in six hypothyroid female patients before and after achievement of a stable euthyroid status. In both conditions, UNSR measured at intervals in response to constant alanine infusion was linearly related to the average alpha-amino N concentrations. In the hypothyroid state, peak UNSR was decreased by 31% in comparison with values measured in euthyroidism, which were in the normal range. FHNC (ie, the slope of the linear relation between UNSR and blood alpha-amino N concentration) is a measure of the kinetics of the process of hepatic amino N to urea N conversion; it was 19.8 +/- 4.0 L.h-1 in hypothyroid patients and increased to normal values after L thyroxine replacement (30.4 +/- 3.3 L.h-1, P < .01; normal values > 25 L.h-1). Hepatic microsomal and cytosolic activities (antipyrine clearance and galactose elimination) were normal in hypothyroid patients and did not change significantly after therapy. Our data show a specific defect in hepatic handling of amino acids in hypothyroid patients, leading to reduced alpha-amino N to urea N conversion, in the absence of any detectable impairment in different hepatic metabolic activities. PMID- 8412739 TI - Indices of thyroid function and weight loss in human immunodeficiency virus infection and the acquired immunodeficiency syndrome. AB - Abnormalities of thyroid hormone levels have been reported in the acquired immunodeficiency syndrome (AIDS), but there has been debate as to whether they are appropriate for the clinical status of the patients. Inappropriate maintenance of circulating 3,3',5-triiodothyronine (T3) levels could contribute to weight loss. Although many patients with AIDS have a history of wasting, recent data indicate that prolonged periods of stable weight occur in AIDS and that short-term weight loss is present in a subset of patients with anorexia, many of whom have active secondary infection (AIDS-SI). Therefore we analyzed thyroid hormone levels in a cohort of subjects that have been characterized in terms of recent weight loss and caloric intake. Asymptomatic patients with human immunodeficiency virus infection (HIV+) had short-term stable weights, normal caloric intake, and normal serum T3 levels. In AIDS, average short-term weight was stable, caloric intake was normal, and T3 levels were decreased by 19%. In AIDS-SI, both short-term weight loss and anorexia were significant, and this group showed a 45% decrease in T3 levels. The free T3 (FT3) index was decreased by 30% in AIDS and by 50% in AIDS-SI. Free thyroxine (FT4) levels were decreased while thyroxine-binding globulin (TBG) capacity was increased in HIV+ and AIDS; TBG sialylation was unchanged. Thyrotropin (TSH) levels were slightly increased in AIDS, although levels remained within the normal range. 3,3',5' triiodothyronine (rT3) levels were decreased in HIV+, AIDS, and AIDS-SI. Thus asymptomatic patients with HIV infection whose weight is stable maintain normal T3 levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412740 TI - The Cincinnati Myocardial Infarction and Hormone Family Study: family resemblance for dehydroepiandrosterone sulfate in control and myocardial infarction families. AB - Dehydroepiandrosterone sulfate (DHEAS) was examined in random (control) and nonrandom (case) families participating in the Cincinnati Myocardial Infarction and Hormone (CIMIH) family study. The case families were ascertained through white men who survived a myocardial infarction (MI) before the age of 56, whereas control families were recruited through advertisements and through an adolescent boy maturation study. Both familial correlations and genetic effects of DHEAS were investigated. First, maximum likelihood estimates of the sex-specific familial correlations (corrected for nonrandom ascertainment) suggested that there was significant heterogeneity between the two sampling types. This heterogeneity was isolated to the male sibling correlation, which was higher in the case than control families. Post hoc analyses suggested that the sibling group heterogeneity may be in part a function of age, since the control sample offspring were on average much younger than those in case families. No sex differences other than those for the siblings were noted in the familial correlations. Second, heritability was investigated in control families using a simple path model (TAU) that allowed for sex differences. The only significant model parameter was the sex-specific familiarity (combined polygenic and familial environmental effects), which was larger in females (74%) than in males (29%). In general, these analyses suggested that (1) DHEAS may play only a limited role in the increased risk for premature MI, and (2) the degree of heritable (familial) variation may be dependent on sex. PMID- 8412741 TI - Evidence that growth hormone stimulates protein kinase C activity in isolated rat hepatocytes. AB - The mechanism of action of growth hormone (GH) is not known, although indirect evidence suggests that protein kinase C (PKC) might play an important role in the insulin-like actions of GH. In this investigation, we directly examined the effects of GH relative to those of insulin on PKC activity in isolated rat hepatocytes. Human GH (10(-7) mol/L) significantly increased the activity of PKC in both cytosolic and particulate fractions. The effect was maximal at 1 minute, disappeared at 5 minutes, and then increased again at 30 minutes in both fractions. At 1 minute, maximal and half-maximal stimulation of PKC activity occurred at hGH concentrations of 10(-7) and 5 x 10(-9) mol/L, respectively. Insulin (10(-7) mol/L) also induced a significant and transient increase in enzyme activity at 2 minutes in cytosolic and particulate fractions; at 30 minutes, PKC activity was decreased in the soluble fraction (-17%) and increased in the particulate fraction (+65%). Measurement of specific [3H]-phorbol dibutyrate (PDBu) binding suggested translocation of PKC from the cytosol to the membrane fraction after 30 minutes of incubation, only after insulin treatment. The early effects of GH and insulin on PKC activity were additive in both the particulate and cytosolic fractions. Although the later effects of GH and insulin on PKC were quite different, both hormones rapidly activated PKC in isolated hepatocytes, suggesting that PKC might be involved in triggering the insulin-like actions of GH. PMID- 8412742 TI - Aluminum accumulation and neurotoxicity in Swiss-Webster mice after long-term dietary exposure to aluminum and citrate. AB - The present study was performed to determine aluminum uptake, retention, and neurotoxic effects in the presence of dietary citrate. Six-week-old female Swiss Webster mice were fed semipurified diets containing 3.5% sodium citrate and either 3 micrograms Al/g diet (3 Al) or 1,000 micrograms Al/g diet (1,000 Al) as AlCl3. After 5 to 7 weeks of feeding these diets, changes in behavior were assessed using the National Institute of Environmental Health Sciences Neurobehavioral Test Battery. Liver and bone Al concentrations in the 1,000 Al group were higher than in the 3 Al group at both the 5- and 7-week time points. Spinal cord Al concentrations in the 1,000 Al group were 200% higher at 5 weeks (P < .01) than in controls, and brain nuclear fraction Al concentrations in the 1,000 Al group were 150% higher at 5 and 7 weeks (P < .01) than in the 3 Al group. The Neurobehavioral Test Battery showed lower grip strength and greater startle responsiveness in the 1,000 Al group compared with the 3 Al group at both the 5- and 7-week time points. Based on reports that Al can act as a pro-oxidant, we examined Al-induced brain lipid and protein oxidative damage; neither was evident in the Al-intoxicated mice. In summary, feeding of Al and citrate to mice resulted in Al accumulation in the central nervous system, and this accumulation was associated with overt signs of neurotoxicity. Brain protein and lipid oxidative damage was not associated with early manifestation of Al toxicity. PMID- 8412743 TI - Muscle group-specific regulation of GLUT 4 glucose transporters in control, diabetic, and insulin-treated diabetic rats. AB - Insulin resistance in diabetic rats involves pretranslational suppression of the GLUT 4 glucose transporter in muscle. Because the capacity for insulin-mediated glucose transport varies as a function of muscle group, we hypothesized that GLUT 4 was differentially expressed and regulated by diabetes in a muscle-specific manner. We studied control (C), streptozocin (STZ)-induced diabetic (D), and insulin-treated diabetic (Tx) rats and examined the following muscles that vary in fiber composition: soleus (type I fibers), gastrocnemius (mixed type IIa > IIb), vastus lateralis and rectus abdominis (type IIb > IIa), and cardiac muscle. In C animals, these muscles exhibited significant differences in the baseline expression of GLUT 4. Relative GLUT 4 content was highest in cardiac muscle, intermediate in soleus, and significantly lower in gastrocnemius, rectus abdominis, and vastus lateralis (1.8:1.0:0.6). The impact of diabetes and insulin therapy on GLUT 4 expression also varied as a function of muscle group. After four weeks of diabetes, GLUT 4 levels were reduced by approximately 50% in cardiac muscle, soleus, and gastrocnemius. In contrast, GLUT 4 content in rectus abdominis and vastus lateralis was similar to that in control rats. Exogenous insulin treatment of diabetic rats increased GLUT 4 content in soleus, cardiac muscle, and gastrocnemius, but had no effect in either vastus lateralis or rectus abdominis. Temporal effects of diabetes and insulin treatment were also examined in different skeletal muscle. Soleus showed a significant decrease in GLUT 4 content as early as 2 days with a further decrease at 4 weeks; rectus abdominis showed little change at either time point.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412744 TI - Phenylalanine conversion to tyrosine: comparative determination with L-[ring 2H5]phenylalanine and L-[1-13C]phenylalanine as tracers in man. AB - The in vivo rate of conversion of phenylalanine to tyrosine (PheOH) can be estimated using combinations of stable isotope-labeled phenylalanine and tyrosine. We have compared in four healthy adult men the rates of phenylalanine conversion to tyrosine based on the following pairs of primed, continuous tracer infusions administered simultaneously: (1) L-[ring-2H5]phenylalanine and 2H2 tyrosine with a 2H4-tyrosine prime, and (2) L-[1-13C]phenylalanine and 2H2 tyrosine with a 1-13C-tyrosine prime. Phenylalanine oxidation was determined from measurement of 13CO2 excretion in expired air. Tracers were given for 8 hours, with subjects being in the postabsorptive state during the first 3 hours and in the fed state during the remaining 5 hours. Mean (+/- SD) rates (mumol.kg-1.h-1) of phenylalanine conversion to tyrosine for fasted and fed states, respectively, were 5.1 +/- 2.9 and 6.8 +/- 3.4 with 2H5-phenylalanine and significantly higher (P < .05) at 11.1 +/- 5.6 and 12.7 +/- 7.7 with 13C-phenylalanine as tracer. Phenylalanine oxidation was 9.9 +/- 2.0 and 13.5 +/- 2.6, respectively, for fasted and fed states, and these mean values did not differ (P > .1) from the rate of phenylalanine conversion to tyrosine determined using 13C-phenylalanine. These results indicate the need for caution in interpreting kinetic aspects of phenylalanine metabolism when based on isotopic data from multideuterated phenylalanine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412745 TI - Alteration of lipoprotein(a) concentration with glycemic control in non-insulin dependent diabetic subjects without diabetic complications. AB - Recently, a high plasma level of lipoprotein(a) [LP(a)] has been considered an independent risk factor for atherosclerosis and its sequelae, particularly myocardial infarction. Patients with non-insulin-dependent diabetes mellitus (NIDDM) have an increased mortality rate from cardiovascular and cerebrovascular disease. Therefore, plasma concentrations of Lp(a) were determined and the relationship between fasting plasma Lp(a) level and diabetic control was investigated in NIDDM patients without any diabetic complications. Fasting plasma Lp(a) levels were measured using enzyme-linked immunosorbent assay kits [Terumo Medical Corp, Elkton, MD, Lp(a)] in 61 NIDDM subjects (30 men aged 56 +/- 2.0 years, 31 women aged 53 +/- 2.1 years [mean +/- SEM]) who were without any diabetic macroangiopathy and microangiopathy such as retinopathy, nephropathy, and neuropathy and in 56 healthy age- and sex-matched controls. Plasma Lp(a) levels were significantly higher in the diabetic group than in the control group (23.5 +/- 2.5 v 11.7 +/- 1.4 mg/dL [mean +/- SEM], P < .001). There was no significant correlation between log-transformed plasma Lp(a) levels and other factors such as age, sex, body mass index (BMI), blood pressure, duration of diabetes, fasting plasma glucose (FPG) level, glycosylated hemoglobin (HbA1C) level, and plasma lipid levels except for low-density lipoprotein cholesterol (LDL-C) levels in diabetic patients. A significant positive correlation was noted in diabetic patients between the changes of log Lp(a) and HbA1C levels after a 3 month follow-up period (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412746 TI - Serum lipoprotein(a) levels differ in different phenotypes of primary hyperlipoproteinemia. AB - Previous studies have indicated that serum lipoprotein(a) [Lp(a)] levels are mostly under genetic control. We have attempted to determine whether serum Lp(a) levels differ in different phenotypes of primary hyperlipoproteinemia (HL). A total of 129 subjects with HL (three with type I, 43 with familial hypercholesterolemia [FH], 17 with type IIa [non-FH], 11 with type IIb, six with type III [E2/2], 44 with type IV, and five with type V) and 18 normolipidemic controls were included in the study. Thirty-two FH subjects were being treated with hypolipidemic agents, but none of the other subjects were receiving any medication. Fasting blood samples were collected for determination of both serum lipid and Lp(a) levels. Lp(a) level was measured by enzyme-linked immunosorbent assay. The 18 controls had serum Lp(a) concentrations of 18.0 +/- 14.5 mg/dL (mean +/- SD), and four of them had high serum Lp(a) levels (> or = 25 mg/dL). Serum Lp(a) concentrations in FH subjects tended to be higher than in the controls (30.5 +/- 25.0 mg/dL), and the incidence of high Lp(a) levels in FH subjects was significantly higher than in the controls (51% v 22%, P < .01). There was no difference between serum Lp(a) levels of FH subjects depending on whether they were receiving medication. In contrast, most of the subjects with selective hypertriglyceridemia had very low serum Lp(a) levels (1.5 +/- 0.7, 8.1 +/- 8.3, and 3.5 +/- 5.3 mg/dL in type I, IV, and V, respectively; P < .01 v controls).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412747 TI - Calcium and sodium handling during volume expansion in essential hypertension. AB - To evaluate the actual role of extracellular fluid volume (ECFV) expansion per se in modulating the rate of urinary calcium excretion, a thermoneutral water immersion (WI) study was conducted in 10 normal subjects and 30 patients with essential hypertension. Central hypervolemia by 2 hours of WI caused a significant diuretic and natriuretic response (P < .005) in normal subjects; no significant changes were detected in urinary calcium and magnesium excretion. WI provoked either an appropriate or exaggerated natriuresis (P < .001) in 21 hypertensive patients; these subjects also exhibited a highly positive correlation between urinary sodium and calcium excretion during WI (P < .001). In the remaining nine hypertensive patients, WI produced a significant diuretic response, but a barely discernible (P = NS) natriuresis (inappropriate response). These subjects also exhibited a significant reduction of urinary calcium (P < .001) and magnesium (P < .01) excretion. The data indicate that (1) volume expansion per se may have a role in regulating calcium excretion in hypertensive subjects; (2) a calcium leak may be attributable to a close relationship between urinary sodium and calcium metabolism, and causally related to a disturbance of sodium and volume homeostasis in hypertension. PMID- 8412748 TI - Restoration of nitrogen homeostasis in a man with ornithine transcarbamylase deficiency. AB - We evaluated the hypothesis that sodium phenylbutyrate-induced phenylacetylglutamine biosynthesis in a man with partial ornithine transcarbamylase (OTC) deficiency has a dual effect; it provides an additional vehicle for waste nitrogen excretion, and in the process it suppresses the patient's residual urea N synthesis, which then may be available for N homeostasis if the need arises. A 38-year-old man was studied over three periods. Period I was a control period during which he received a fixed caloric and N intake plus L-citrulline. Phenylbutyrate was added in period II and was maintained during period III, during which his N intake was increased. Plasma levels of ammonium and glutamine and net urea N synthesis were measured in each period; phenylacetylglutamine N synthesis was measured in periods II and III. These studies demonstrated that phenylbutyrate administration led to a 73% decrease in net de novo urea N synthesis during period II, which subsequently increased threefold in period III in response to the increased N intake. Phenylacetylglutamine N synthesis was 2.27 g/d, similar to his estimated maximum net urea N synthesis of 2.65 g/d. During periods II and III, his plasma levels of ammonium and glutamine improved as compared with period I when they were abnormally high. We conclude that sodium phenylbutyrate treatment of patients with urea cycle disorders who have significant residual enzyme activity results in both an improvement in waste N excretion and improved N homeostasis as a result of the generation of a reserve urea N synthetic capacity. This therapeutic approach may be useful in other nitrogen accumulation decreases, eg, portal systemic encephalopathy. PMID- 8412749 TI - The body composition and lipid metabolic effects of long-term ethanol feeding during a high omega 6 polyunsaturated fatty acid diet in micropigs. AB - Our previous research with miniature pigs has shown that long-term ethanol feeding with a low-fat diet decreases arachidonic acid (20:4 omega 6) levels in multiple tissues, but we did not find significant liver pathology. In this study, we investigated the effect of ethanol feeding with high dietary linoleic acid (18:2 omega 6) on tissue fatty acid (FA) profiles and body composition. Five Yucatan micropigs were fed 370 kJ (89 kcal)/kg body weight of a diet containing ethanol and fat as 40% and 34% of energy, respectively; five control pigs were pair-fed corn starch in place of ethanol. Corn oil, 61% 18:2 omega 6, supplied most of the dietary fat. Liver biopsies were performed at baseline (n = 2 per group) and at three other time points (n = 5 per group). Phospholipid (PL) FA levels were measured by thin-layer and gas chromatography. Body composition was analyzed by underwater weighing of carcasses. Body composition analysis demonstrated a marked reduction of carcass fat in the ethanol group, but no significant reduction of carcass lean weight after 12 months. In liver PLs, the ethanol group showed decreased 20:4 omega 6 and docosahexaenoic acid (22:6 omega 3) after 1 month. While the decreased 20:4 omega 6 remained constant after 1 month, 22:6 omega 3 showed a progressive decrease up to 12-months, resulting in a continuous decrease of the omega 3/omega 6 FA ratio. This slowly progressive decrease in the omega 3/omega 6 ratio in liver PLs with ethanol feeding may have enhanced the inflammatory response in the liver, contributing to liver pathology. Body composition results indicate marked wasting of energy in the ethanol group. PMID- 8412750 TI - Effect of high-dose endotoxin on glucose production and utilization. AB - The purpose of the present study was to determine how a high dose of endotoxin (lipopolysaccharide [LPS]), which produces hypoglycemia, alters in vivo glucose uptake by individual tissues. Catheterized conscious fasted rats were injected intravenously (i.v.) with either saline, LPS (1 mg/100 g body weight [BW], lethal dose [LD] 100), or 3-mercaptopicolinic acid (3-MP), an inhibitor of gluconeogenesis. In the latter two groups, blood glucose levels were clamped at either 6 mmol/L (euglycemia) or 3 mmol/L (hypoglycemia). In the first series of experiments, whole-body glucose flux was determined using [3-3H]glucose, and in the second study in vivo glucose uptake (Rg) by individual tissues was estimated by the tracer [U-14C]-2-deoxyglucose technique. The relative contribution of hypoglycemia per se to the LPS effect was determined by comparing the values from LPS- versus 3-MP-treated animals. There was no difference in the rate of whole body glucose utilization (Rd) between saline-infused control rats and LPS-treated animals that were hypoglycemic. However, Rg by diaphragm, spleen, liver, and lung was increased in hypoglycemic LPS-treated rats. The increased Rg in these tissues was not observed in 3-MP-treated rats with a comparable hypoglycemia. Only the gastrocnemius muscle showed a reduction in Rg under hypoglycemic conditions, and the decrease was similar in both LPS- and 3-MP-treated animals. When sufficient glucose was infused into LPS-injected rats to maintain euglycemia, whole-body glucose Rd was increased compared with that in hypoglycemic LPS-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412751 TI - Insulin sensitivity in familial hypercholesterolemia. AB - Insulin resistance is found in association with obesity, non-insulin-dependent diabetes mellitus, and essential hypertension, which are all risk factors for atherosclerotic cardiovascular disease. Furthermore, hyperinsulinemia has been reported in familial combined hyperlipoproteinemia and endogenous hypertriglyceridemia. Finally, relatively high serum triglyceride and low high density lipoprotein (HDL) cholesterol concentrations invariably accompany hyperinsulinemia. Whether insulin sensitivity is affected by the isolated presence of high levels of serum low-density lipoprotein (LDL) cholesterol has not been clearly established. We studied 13 subjects with heterozygous familial hypercholesterolemia (FHC) and 15 normocholesterolemic subjects selected to be free of any other known cause of insulin resistance. Thus FHC patients and controls had normal body weight and fat distribution, glucose tolerance, blood pressure, and serum triglyceride and HDL cholesterol concentrations, but were completely separated on plasma LDL cholesterol concentrations (6.05 +/- 0.38 v 3.27 +/- 0.15 mmol/L, P < .0001). Fasting plasma levels of glucose, insulin, free fatty acids (FFA), and potassium and fasting rates of net carbohydrate and lipid oxidation were superimposable in the two study groups. During a 2-hour euglycemic (approximately 5 mmol/L) hyperinsulinemic (approximately 340 pmol/L) clamp, whole body glucose disposal rates averaged 30.4 +/- 2.3 and 31.1 +/- 3.0 mumol.kg-1 x min-1 in FHC and control subjects, respectively (P = 0.88). The ability of exogenous hyperinsulinemia to stimulate carbohydrate oxidation and energy expenditure and suppress lipid oxidation and plasma FFA and potassium levels was equivalent in FHC and control subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412752 TI - Effects of thiazolidinediones on glucocorticoid-induced insulin resistance and GLUT4 glucose transporter expression in rat skeletal muscle. AB - Thiazolidine-2,4-diones, a new class of oral antihyperglycemic agents, have been shown to be effective in improving insulin sensitivity in a number of animal models of insulin resistance, and recent investigation has suggested that the mechanism of action of these agents may include upregulation of the GLUT4 (insulin-regulatable) glucose transporter. We studied the efficacy of two of these agents, pioglitazone and englitazone, in preventing glucocorticoid-induced insulin resistance in rats, and examined the potential role of changes in GLUT4 expression in their action in skeletal muscle. Rats were treated with 0.1 mg/d dexamethasone for 6 to 7 days with or without either pioglitazone (10 mg/kg/d) or englitazone (50 mg/kg/d). Both thiazolidinediones decreased the elevated fasting serum glucose and insulin levels in dexamethasone-treated animals. Dexamethasone treatment alone decreased insulin-stimulated 2-deoxyglucose uptake into isolated soleus muscles to 35% of control values. The addition of pioglitazone or englitazone increased insulin-stimulated 2-deoxyglucose uptake by 74% and 57%, respectively. Whereas dexamethasone treatment alone increased GLUT4 protein content in rat soleus muscle by 25%, additional treatment with pioglitazone or englitazone did not further significantly alter GLUT4 levels. We conclude that thiazolidinediones enhance insulin responsiveness in skeletal muscle during glucocorticoid treatment, but their mode of action in this setting is not via upregulation of GLUT4 expression. PMID- 8412753 TI - Acute reduction of renal 11 beta-hydroxysteroid dehydrogenase activity by several antinatriuretic stimuli. AB - The 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) activity of the kidney prevents access of cortisol or corticosterone to the renal mineralocorticoid receptor. Reduction of 11 beta-HSD activity by nutritional, hormonal, or pharmacologic factors might enhance the mineralocorticoid effect of these corticosteroids, thus causing sodium retention. To test this concept, we studied the effect on 11 beta-HSD activity of several antinatriuretic factors given orally to rats or exposed in vitro to rat renal tissue. Renal 11 beta-HSD activity was higher in fasted than fed rats (P < .05). Glucose, ethanol, and Toradol (Syntex Laboratories, Palo Alto, CA) given orally to fasted rats all reduced renal 11 beta-HSD activity by 20% to 40% (P < .05-.005) to levels similar to those observed in fed animals. Incubation of renal tissue from fasted rats with physiologic concentrations of insulin, ethanol, and Toradol also reduced 11 beta-HSD activity by 20% to 40% (P < .05-.01). These findings are consistent with the hypothesis that the antinatriuretic actions of these stimuli are due in part to alteration of renal 11 beta-HSD leading to greater mineralocorticoid effects in kidney. PMID- 8412754 TI - Compensatory increase in intestinal apolipoprotein A-IV mRNA levels in the experimental nephrotic rat. AB - Using experimental nephrotic rats, we investigated the potential feedback regulation of apolipoproteins (apos) at their hepatic and intestinal synthetic sites. Nephrotic syndrome (NS) was induced in rats by puromycin aminonucleoside (PAN) with a single intraperitoneal injection (120 mg/kg). In nephrotic rats, we observed a 60% reduction in serum apo A-IV levels despite a 3.4-fold increase in jejunum and a 1.5-fold increase in ileum apo A-IV mRNA levels, although hepatic apo A-IV levels were unchanged compared with those in pair-fed control rats. A strikingly positive correlation was observed between daily urinary excretion of apo A-IV and its mRNA levels in jejunum (r = .856, P < .01; n = 10) and ileum (r = .710, P < .05; n = 10). On the other hand, nephrotic rats had an 8.2-fold increase in serum apo A-I level associated with a 4.6-fold increase in hepatic and a small but significant increase in jejunum apo A-I mRNA levels. Compared with the fractional catabolic loss of albumin or apo A-IV, that of apo A-I was small and suggests a diminished level of glomerular filtration, leading to a further elevation in serum apo A-I level. Barring nonspecific effects of PAN, these data suggest that reduction of serum apo A-IV level due to urinary loss may directly upregulate mRNA levels in the small intestine. Alternatively, it may be the result of an effective filtration of a serum component unassociated with lipoproteins that normally and site-specifically reduces apo A-I and apo A-IV mRNA transcription. PMID- 8412755 TI - Dichloroacetic acid accelerates initial development of 2-cell murine embryos in vitro. AB - Preimplantation embryos up to the 8-cell stage of development use lactate and pyruvate but not glucose or Krebs cycle intermediates to support growth, development, and cleavage. The dominant effect of dichloroacetic acid (DCA) is the irreversible stimulation of pyruvate dehydrogenase (PDH) activity, thus accelerating the oxidative metabolism of pyruvate and lactate. To test the hypothesis that early induction of oxidative metabolism in 2-cell murine embryos accelerates preimplantation embryo cleavage rates, female B6C3F1 mice at 6 to 8 weeks of age were superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) and mated. All 2-cell stage embryos were randomly assigned to culture media with or without 130 micrograms/mL DCA. The developmental stage of all embryos was then noted every 24 hours for a total of 72 hours. Chi-square analysis and the method of average rank sum were used to compare the distribution of embryos at each observation point. At 24 hours, DCA exposed embryos had achieved an advanced stage of growth and development relative to controls (average rank sum, P = .026; chi-square distribution, P = .047). Subsequently, at 48 and 72 hours, neither the average rank sum nor the chi-square distribution was different. Our data suggest that DCA accelerates early growth and development of murine embryos before implantation, possibly through the early induction of oxidative metabolism. PMID- 8412756 TI - Insulin influences immunoreactive endothelin release by human vascular smooth muscle cells. AB - This study sought to investigate whether insulin influences immunoreactive endothelin release from cultured human vascular smooth muscle cells. For this purpose, we incubated cultured vascular smooth muscle cells (VSMC) obtained from human microvessels with 0, 80, and 320 microU/mL insulin with or without arginine vasopressin (10 nmol/L) and angiotensin II (10 nmol/L). After 6 hours, the culture supernatant was collected and immunoreactive endothelin was determined by radioimmunoassay. Insulin at a concentration of 320 microU/mL induced a significant increase of immunoreactive endothelin levels in medium (from 15.2 +/- 0.8 to 20.6 +/- 0.8 pg/200 microL, P < .01) and potentiated arginine vasopressin- and angiotensin II-induced immunoreactive endothelin release (P < .0001 and P < .04, respectively). Insulin at a concentration of 80 microU/mL did not induce a significant increase of spontaneous immunoreactive endothelin release, but significantly increased the effects of arginine vasopressin (P < .05). In conclusion, insulin influences immunoreactive endothelin release from human VSMC in culture. PMID- 8412757 TI - Leucine metabolism during chronic ethanol consumption. AB - Chronic ethanol consumption is known to increase plasma concentrations of branched-chain amino acids (BCAA) in rats and man, but the mechanisms of this effect are not known. Chronic ethanol consumption may increase levels of BCAA by altering protein turnover and/or by affecting the oxidation of BCAA. These possibilities were investigated in rats pair-fed liquid diets containing either 0% or 36% of total calories as ethanol for 21 days. In the fed state, ethanol treated rats had a plasma ethanol level of 20 +/- 5 mmol/L and twofold increases in BCAA concentrations in plasma. There were also significant increases (37% to 63%) in muscle, liver, and jejunal mucosa BCAA concentrations. Chronic ethanol consumption significantly increased whole-body rates (mumol/100 g/h) of leucine turnover (73.8 +/- 7.5 v 104 +/- 5.6, P < .01) and oxidation (12.0 +/- 1.7 v 17.7 +/- 1.1, P < .05). In contrast, it significantly decreased leucine incorporation (nmol/mg protein/240 min) into both muscle (0.61 +/- 0.07 v 0.35 +/- 0.05, P < .01) and liver (13.25 +/- 1.40 v 6.78 +/- 0.98, P < .01) proteins. Incorporation of leucine into the mucosal proteins of jejunum (17.42 +/- 1.42 v 15.85 +/- 1.90, P = NS) was not significantly altered by ethanol. These results suggest that reduced protein synthesis and/or increased protein breakdown may account for the elevated tissue BCAA concentrations in chronic ethanol consumption. The consequences of these increased tissue concentrations are increases in tissue oxidation and plasma concentrations of BCAA. PMID- 8412758 TI - Sodium-lithium countertransport activity is not affected by short-term insulin exposure in vivo or in a physiologic medium in vitro. AB - This study examined the acute effects of physiologic concentrations of insulin in vitro and in vivo on sodium-lithium countertransport (SLC) kinetics in nondiabetic subjects. SLC was measured at eight different external sodium concentrations including the standard 150 mmol/L, allowing calculation of both maximal velocity (Vmax) and external sodium affinity (Km). Incubation with insulin (50 mU/L) in 110 MgCl2 but not in 150 mmol/L NaCl decreased standard SLC activity. The decrease was accounted for by a reduction in Vmax, whereas Km remained constant. There was no difference in standard SLC activity, Vmax, or Km when endogenous insulin concentrations were altered either by fasting or by a carbohydrate load. Similarly, standard SLC activity, Vmax, or Km were not significantly different before or at the completion of a euglycemic hyperinsulinemic clamp. These findings provide no support for an action of physiologic concentrations of insulin either in vitro or in vivo on the kinetics of the countertransporter measured in vitro in isotonic sodium-containing media. PMID- 8412759 TI - Insulin resistance, glucose intolerance, and hyperinsulinemia in patients with microvascular angina. AB - Plasma glucose and insulin responses to oral glucose and insulin-mediated glucose disposal were determined in 20 patients with microvascular angina and 20 normal volunteers who were similar in terms of age, gender distribution, and degree of obesity. Plasma glucose and insulin responses to a 75-g oral glucose challenge were significantly higher in those with microvascular angina (P < .001), as were steady-state plasma glucose concentrations after a 180-minute infusion of somatostatin, glucose, and insulin (12.2 +/- 1.0 v 7.6 +/- 0.6 mmol/L, P < .001). Since steady-state plasma insulin concentrations were similar in the two groups (627 +/- 32 v 631 +/- 29 pmol/L), these data indicate that patients with microvascular angina are insulin-resistant, glucose-intolerant, and hyperinsulinemic compared with a matched group of normal volunteers. PMID- 8412760 TI - Insulin-like growth factors and non-islet cell tumor hypoglycemia. PMID- 8412761 TI - Reduced tissue thyroid hormone levels in fatal illness. AB - Patients with severe nonthyroidal illnesses (NTIs) frequently have decreased serum concentrations of triiodothyronine (T3) and less often of thyroxine (T4) without clear evidence of hypothyroidism. To determine whether T3 and T4 levels are also reduced in the tissues, we analyzed autopsy samples from 12 patients dying of NTI and 10 previously healthy individuals dying suddenly from trauma. Mean serum T3, T4, and free T4 index values were lower by 79%, 71%, and 49%, respectively, in the NTI group than in controls, but serum thyrotropin (TSH) values did not differ significantly. Mean T3 concentrations in cerebral cortex, hypothalamus, pituitary, liver, kidney, and lung were lower in the NTI group than in controls by 43% to 76%, but mean values in heart and skeletal muscle did not differ significantly between the groups. The mean liver T4 concentration was 66% lower in the NTI group, but mean T4 concentrations in the cerebral cortex were similar in the two groups. These results indicate that many tissues may be deficient in thyroid hormones in patients with fatal NTI, although the severity of the reduction in thyroid hormone concentrations may vary from one organ to another. PMID- 8412762 TI - Alanine uptake by liver of mid-lactating rats. AB - L-Alanine transport in liver plasma membrane vesicle preparations from fed virgin and 15-day-lactating rats was studied. Lactation was found to induce a decrease of the maximal rate (Vmax) of a high-capacity-low-affinity component of the Na(+) dependent L-alanine uptake. However, a high-affinity-low-capacity agency was significantly induced in lactating-rat livers. L-Alanine uptake was differentially inhibited by other amino acids in those preparations from lactating rats, and showed different sensitivity to Li+ as a cosubstrate instead of Na+ and to inhibition by sulfhydryl modifying reagents (N-ethylmaleimide [NEM] and p-chloromercuribenzosulfonate [PCMBS]). All of these observations taken together suggest that system A is upregulated in lactating-rat livers, thus resulting in a different contribution of both agencies A and ASC to the total Na(+)-dependent alanine transport into liver plasma membrane vesicles. This was demonstrated using the analogue alpha-methyl-aminoisobutyric acid (MeAIB), a specific system A substrate. L-Alanine uptake rates, as calculated from plasma membrane enzyme marker recoveries, were also enhanced in the physiologic range of alanine concentrations in blood. Our results prove that the physiologic adaptation to lactation involves modulation of system A activity in the liver. PMID- 8412763 TI - Prednisolone enhances beta-cell function independently of ambient glycemic levels in type II diabetes. AB - This study examined the changes in beta-cell response and insulin sensitivity induced by a single overnight dose of 15 mg prednisolone in eight type II diabetic subjects, seven nondiabetic normal controls, and eight subjects with a first-degree type II diabetic relative who were therefore at risk of developing diabetes. beta-Cell secretion was assessed by use of the hyperglycemic clamp technique, and insulin sensitivity was assessed with the clamp and the Continuous Infusion of Glucose with Model Assessment (CIGMA) technique. Subjects were studied in random order on two occasions, after placebo and after prednisolone administration. Normal subjects showed an increase of median fasting glucose level from 4.7 to 5.2 mmol.L-1 after prednisolone (P < .02) and at-risk subjects showed an increase from 4.8 to 5.5 mmol.L-1 (P < .005), whereas diabetic subjects showed no significant increase in median fasting plasma glucose level (7.0 mmol.L 1 after placebo and 6.3 mmol.L-1 after prednisolone). Six of these eight diabetic subjects showed a paradoxical decrease of fasting plasma glucose level after prednisolone therapy. All three groups showed a significant elevation of clamp steady-state plasma insulin levels following prednisolone, with a median percentage elevation of 46%, 66%, and 31% for normal, at-risk, and diabetic subjects, respectively. All three groups showed significant reduction in insulin sensitivity measured by CIGMA following prednisolone of 51%, 41%, and 25% of pre prednisolone levels in normal, at-risk, and diabetic subjects, respectively, with a significantly greater reduction in normal subjects than in diabetics (P < .02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412764 TI - Effect of medium pH on glutathione redox cycle in cultured human umbilical vein endothelial cells. AB - Impairments of the glutathione redox cycle in cultured endothelial cells under acidic pH conditions were measured. Glutathione-dependent H2O2-degrading activities decreased by 20% (P < .01) at pH 6 and by 51% (P < .01) at pH 4 compared with activities at pH 7.4 1 hour after a change with fresh medium. Intracellular reduced glutathione (GSH) content increased by 85% (P < .01) following the change with pH 7.4 medium. Such increases in GSH content were impaired after exposure to acidic medium. After exposure to 500 mumol/LH2O2, intracellular GSH content decreased by 61% compared with the level obtained in the absence of H2O2 at pH 7.4 (P < .01). Compared with the level at pH 7.4, the H2O2-induced decrease in intracellular GSH content was 32% lower (P < .01) at pH 6 and did not change at all at pH 4. After exposure to 500 mumol/L H2O2, the intracellular oxidized glutathione (GSSG) content increased by 160% at pH 7.4 (P < .01), 370% at pH 6 (P < .01), and 90% at pH 4 compared with treatment without H2O2, respectively. After exposure to 500 mumol/L H2O2, the release of GSSG from cells at pH 6 decreased by 38% compared with the value found at pH 7.4 (P < .05), and the release at pH 4 completely disappeared. Both glutathione peroxidase (GPO) and glutathione reductase activities decreased as a function of a decrease in pH from 7.4 to 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412765 TI - Abnormal polyunsaturated lipid metabolism in the obese Zucker rat, with partial metabolic correction by gamma-linolenic acid administration. AB - Below-normal proportions of phospholipid (PL) arachidonic acid (20:4 omega 6) have been reported in serum from obese humans and in liver from obese Zucker rats. This implies an abnormality of 20:4 omega 6 formation from linoleic acid (18:2 omega 6), possibly in the delta 6 desaturase step, or alternatively an abnormality in the catabolism or distribution of arachidonate. We previously speculated that a reduced proportion of 20:4 omega 6 in hepatic PL could contribute to the etiology of genetic obesity. Providing 18:3 omega 6 would bypass delta 6 desaturase and possibly normalize hepatic PL 20:4 omega 6. Therefore weanling Zucker rats were given free access to a defined diet (11% of energy as soy oil) and gavaged daily with 100 microL of either black currant oil concentrate ([BCO] 8% 18:2 omega 6 and 70% 18:3 omega 6) or soy oil ([Soy] 55% 18:2 omega 6 and < 0.1% 18:3 omega 6). Groups of eight lean and eight obese animals were randomized to receive Soy or BCO in a 2 x 2 design; 10 obese and 10 lean rats were fed a stock diet (nongavaged reference). All groups of lean rats had identical weight gain; food intake for Soy lean and BCO lean did not differ. The obese reference animals and Soy obese animals did not differ in weight gain. However, BCO obese animals ate less food (P < .06), gained less weight (P < .0001), and had lower percent body fat (P < .05) compared with the Soy obese animals. The fatty acid constituents from serum, liver, and adipose tissue showed marked differences between lean and obese animals. Hepatic PL 20:4 omega 6 was lower in Soy obese than in lean (P < .002), but was normalized by BCO gavage (diet effect, P < .007). The paucity of hepatic PL 20:4 omega 6 was not due to reduced desaturase activity, as the proportions of other desaturase products (20:3 omega 6, 20:3 omega 9, 20:5 omega 3) were significantly elevated in Soy obese rat liver and serum. Serum and hepatic cholesteryl ester 20:4 omega 6 levels were elevated in obese versus lean rats (P < .02 and P < .0001), indicating abnormal arachidonate distribution in the obese Zucker rat. Because BCO selectively reduced weight gain and percent body fat in obese Zucker rats, our results imply a role for abnormal omega 6 fatty acid metabolism in the etiology of Zucker obesity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8412766 TI - Adrenal steroid and adrenocorticotropin responses to human corticotropin releasing hormone stimulation test in adolescents with type I diabetes mellitus. AB - To determine whether abnormalities of hypothalamic-pituitary-adrenal axis function occur in type I diabetes mellitus, corticotropin, cortisol, 17 hydroxyprogesterone (17-OHP), androstenedione (D4-A), dehydroepiandrosterone (DHEA), and DHEA sulfate (DS) levels were measured after an intravenous (IV) injection of 1 microgram/kg human corticotropin-releasing hormone (CRH) in diabetic adolescents and normal age-matched subjects. CRH produced a consistent increase in corticotropin blood levels that was comparable in the two groups. In contrast, both baseline and stimulated cortisol concentrations were greater in diabetic patients. Levels of 17-OHP increased after CRH administration, and the magnitude of increase was similar in all subjects. Stimulation with CRH determined an attenuated integrated DS response in diabetics compared with normal subjects with a different pattern of the hormone secretion, whereas no differences in D4-A concentrations were detected between the two groups. DHEA serum levels of subjects from both groups underwent similar changes following administration of CRH. In conclusion, patients with type I diabetes have a discrete response of adrenal steroids to CRH stimulation that appears to be independent of corticotropin secretion. This phenomenon might be related to a direct effect of insulin on enzyme systems involved in the biosynthetic pathway of adrenal steroids or, alternatively, to an intra-adrenal CRH/corticotropin mechanism acting on the adrenal cortex in a paracrine manner. PMID- 8412767 TI - Long-term endocrine function in hypercholesterolemic patients treated with pravastatin, a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor. AB - Steroid hormone production within the gonads and adrenals requires a continuous supply of cholesterol derived from de novo synthesis within the gland and from uptake of circulating plasma lipoproteins. Steroid hormone secretion was prospectively studied over 24 months in 64 hypercholesterolemic subjects (group I, aged 52 +/- 1 years [mean +/- SEM], 61% male) participating in a randomized double-blind clinical trial of pravastatin (20 to 80 mg daily), a new 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, compared with patients taking cholestyramine or other lipid-lowering drugs (group II). Attempts were made in both groups to maintain serum low-density lipoprotein cholesterol (LDL-C) levels between the 25th and 50th percentile for age and gender. At 24 months, serum LDL-C level decreased by 42% +/- 3% in group I and 44% +/- 1% in group II (P < .001 v baseline, NS between groups). Basal secretion of cortisol, aldosterone, and dehydroepiandrostenedione sulfate (DHEA-S) was maintained throughout the study. However, the serum DHEA-S secretory response to Cortrosyn (Organon, West Orange, NJ) diminished in both treatment groups at 6 and 12 months (P < .05). In men, basal serum testosterone levels and the testosterone response to human chorionic gonadotropin (HCG) did not change. There was some diminution of sperm motility noted in both treatment groups at 6 and 12 months in the subset of men undergoing semen analysis (n = 14, P < .05). In conclusion, pravastatin had no significant effect on steroid metabolism. Changes noted in DHEA-S were not specific for pravastatin, suggesting that this impairment is related to lipid lowering effects. PMID- 8412768 TI - Differential effects of estrogen on low-density lipoprotein subclasses in healthy postmenopausal women. AB - The use of estrogen by postmenopausal women decreases plasma low-density lipoprotein (LDL) cholesterol levels. To determine whether LDL subclass profiles influence this response, we studied 31 healthy postmenopausal women who were administered two doses (0.625 and 1.25 mg/d) of conjugated equine estrogen in a placebo-controlled double-blind crossover study. Lipid-stained gradient gels were used to categorize LDL subclass patterns. All women were classified as LDL subclass pattern A (predominant LDL peak > or = 260 A). Within the pattern A classification, there were 12 women during placebo treatment with LDL subclass I pattern (predominant LDL peak > 271 A) and 19 women with LDL subclass II pattern (predominant LDL peak < or = 271 and > or = 260 A). Postmenopausal women with LDL subclass I on placebo treatment had significantly lower LDL cholesterol levels compared with women having LDL subclass II (126 +/- 28 v 147 +/- 23 mg/dL, P < .03). Postmenopausal women with LDL subclass I also had significantly (P < .05) lower very-low-density lipoprotein (VLDL) cholesterol, VLDL triglyceride, and VLDL apo B levels and significantly higher (P < .05) high-density lipoprotein 2 (HDL2) cholesterol, HDL3 cholesterol, and HDL2 apo A-I levels. Estrogen replacement significantly (P < .05) decreased LDL cholesterol levels and increased VLDL and LDL triglyceride, HDL2 and HDL3 cholesterol and apo A-I, and HDL2 apo A-II levels to a similar extent in postmenopausal women with LDL I or II subclass patterns.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412769 TI - Relationship between changes in serum thyrotropin and total and lipoprotein cholesterol with prolonged Antarctic residence. AB - Antarctic residence (AR) is associated with a 50% increase in the thyrotropin (TSH) response to TSH-releasing hormone (TRH) and an expanded triiodothyronine (T3) distribution volume and extravascular hormone pool, collectively called the polar T3 syndrome. To investigate the possible biologic significance of this syndrome, we studied the relationship between nonstimulated TSH and serum lipid profiles in nine subjects, once while in California and monthly during 9 months of AR. We measured serum levels of TSH, total thyroxine (TT4), free T4 (FT4), total T3 (TT3), free T3 (FT3), thyroid-binding globulin (TBG), total cholesterol (T-CHOL), high-density lipoprotein cholesterol (HDL-C), triglyceride (TG), dietary cholesterol (D-CHOL), dietary fat (D-FAT), and dietary kilocalories at each month. The paired mean monthly change from baseline was used to determine significance. The group's mean levels of TSH (approximately 30%), TBG (approximately 16%), T-CHOL (approximately 4%), HDL-C (approximately 10%), and D CHOL (approximately 19%) increased with AR (P < .05). Small but significant decreases (P < .05) were observed in the mean changes of TT4 (approximately 8%), FT4 (approximately 6%), and TT3 (approximately 6%). FT3, D-FAT, dietary kilocalories, body weight, TG, and the calculated low-density lipoprotein (LDL-C) were unchanged with AR. A significant rate of change (P < .05) during AR was also calculated from the slope of a fitted logarithmic function for TSH (0.96 +/- 0.31 mU.L-1 x mo-1), TBG (61.19 +/- 12.29 nmol.L-1 x mo-1), TT3 (0.09 +/- 0.04 nmol.L 1 x mo-1), TT4/TBG (-0.06 +/- 0.01/mo), TT3/TBG (-8.49 +/- 1.98 x 10(-4)/mo), and TG (-0.33 +/- 0.15 mmol.L-1 x mo-1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412770 TI - Insulin sensitivity, insulin secretion, and glucose effectiveness in anorexia nervosa: a minimal model analysis. AB - The aim of the present study was to estimate insulin secretion, insulin sensitivity (SI), and glucose effectiveness (SG) in subjects with anorexia nervosa. Eight nondiabetic anorectic patients who were dietary restricters and 16 age- and sex-matched healthy control subjects without family history of diabetes were studied. They underwent a modified frequently sampled intravenous glucose tolerance test; glucose (300 mg/kg body weight) was administered and insulin (4 mU/kg body weight/min) was infused from 20 to 25 minutes after administration of glucose. SI and SG were estimated by Bergman's minimal model method. Basal glucose (75.5 +/- 2.1 v 87.1 +/- 1.7 mg/dL) and insulin (3.6 +/- 0.4 v 6.3 +/- 0.5 microU/mL) concentrations were significantly lower in anorectic patients than in control subjects (P < .01). No significant difference was observed in glucose disappearance rate (KG) between the anorectic and control subjects (1.56 +/- 0.5 v 2.26 +/- 0.15%/min). Insulin secretion assessed by the integrated area of plasma insulin above basal level during the first 20 minutes after intravenous stimulation with glucose was significantly decreased in anorectic patients (283 +/- 69 microU.mL-1 x min) compared with control subjects (529 +/- 63 microU.mL-1 x min, P < .05). SI was significantly increased in anorectic patients compared with control subjects (11.2 +/- 1.2 v 7.5 +/- 1.0 x 10(-4) min-1 +/-.[microU/mL] 1, P < .05). However, SG was significantly decreased in anorectic patients (0.015 +/- 0.003 min-1) compared with control subjects (0.023 +/- 0.002 min-1, P < .05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412771 TI - Glucagon and noradrenaline reduce erythrocyte deformability. AB - An impairment in red blood cell (RBC) deformability has been reported in several diseases and might be involved in the pathogenesis of atherosclerosis. The aim of the present study was to test in vitro the effect of two "stress hormones," norepinephrine and glucagon, on the filterability of RBCs from healthy subjects. Measurements were performed with the Hanss hemorrheometer, and results were expressed as the rigidity index (RI). All concentrations of norepinephrine from 10(-7) to 10(-4) mol/L induced a significant increase in RI compared with Hanks buffer, with the maximal effect being reached using 10(-6) mol/L. All concentrations of glucagon from 0.01 to 5 ng/mL also induced a significant increase in RI, with the maximal effect being achieved using 1 ng/mL. These data suggest that these two stress hormones could be involved in the rheologic impairments found in several diseases and therefore in the pathogenesis of atherosclerosis. PMID- 8412772 TI - Resting energy expenditure and weight loss in human immunodeficiency virus infected patients. AB - Resting energy expenditure (REE) and body composition were investigated in 60 clinically stable patients with human immunodeficiency virus (HIV) infection varying with respect to immune impairment. REEs differed significantly from predicted values (> or < 10% of the Harris-Benedict [HB] equation) in 40% of patients. Seven percent of patients showed markedly increased REE (> +20% of HB prediction), whereas REE was decreased in 13% (< -10%). Increased REE was found during all clinical stages of the disease (Walter Reed [WR] 2 through 6) and was not strictly associated with the degree of immune impairment, presence of diarrhea or Kaposi's sarcoma, nutritional state, or anamnestic wasting. Twenty seven patients were evaluated for a mean period of 319 days; 11 lost more than 5% of their initial body weight during the observation period. Weight-losing patients were normometabolic before but showed a significantly increased REE (+7% of predicted values or +8% when compared with previous measurements) during weight loss. The degree of deviation from estimated REE was strongly associated with the degree of weight loss. We summarize that increased REE is not a constant feature of HIV infection. It is not associated with clinical and laboratory parameters of immune deficiency, but may occur during weight loss. Thus increased REE represents an inadequate adaptation to malnutrition and contributes to wasting. PMID- 8412773 TI - Reversal of hyperglycemic-induced defects in myo-inositol metabolism and Na+/K+ pump activity in cultured neuroblastoma cells by normalizing glucose levels. AB - myo-Inositol accumulation and incorporation into phosphoinositides was decreased in neuroblastoma cells chronically exposed to medium containing 30 mmol/L glucose or 30 mmol/L galactose. In addition, the intracellular content of myo-inositol and phosphatidylinositol was decreased and the sorbitol or galactitol content increased in cells cultured for 2 weeks in medium containing 30 mmol/L glucose or 30 mmol/L galactose, respectively. Na+/K+ adenosine triphosphatase (ATPase) transport activity was also significantly decreased by long-term exposure of neuroblastoma cells to medium containing 30 mmol/L glucose or 30 mmol/L galactose. When glucose-conditioned cells were placed in medium containing a normal glucose concentration for 24 hours, myo-inositol metabolism and content, phosphatidylinositol levels, and Na+/K+ pump activity were restored or completely returned to normal values. These functions were also significantly improved, except for the phosphatidylinositol content, which was increased by 55%, when galactose-conditioned cells were incubated for 24 hours in unsupplemented medium. The polyol content of the glucose- or galactose-conditioned cells was also significantly reduced. Returning the cells to normal glucose levels for 1 to 3 hours did not completely restore myo-inositol metabolism. Improved myo-inositol metabolism and content, sorbitol levels, and Na+/K+ ATPase transport activity were also obtained within 24 hours when cells chronically exposed to medium supplemented with 30 mmol/L glucose were placed in medium containing 30 mmol/L glucose and 0.4 mmol/L sorbinil. The phosphatidylinositol content of these cells was improved by approximately 30%. Cells prelabeled for 24 hours with [U 14C]sorbitol metabolize more than 50% of the [U-14C]sorbitol during a 24-hour incubation in unsupplemented medium. These studies conducted at the cellular level suggest that restoration of normal myo-inositol metabolism, polyol content, and Na+/K+ pump activity altered by hyperglycemic conditions occurs rapidly following normalization of glucose concentration. PMID- 8412774 TI - Evidence that inhibition of muscle amino acid uptake during endotoxemia is not mediated by glucocorticoids. AB - Sepsis and endotoxemia are associated with increased muscle protein breakdown and inhibited amino acid uptake. Glucocorticoids are important for the regulation of muscle protein breakdown in catabolic conditions; in contrast, the role of glucocorticoids in the regulation of muscle amino acid transport during sepsis or endotoxemia is not known. The present study was designed to test the role of glucocorticoids in the regulation of muscle amino acid uptake during endotoxemia. Amino acid transport, determined as uptake of 3H-alpha-aminoisobutyric acid (AIB) by incubated soleus muscles in vitro, was reduced by approximately 40% 2 hours after intraperitoneal (IP) injection of 10 micrograms/kg endotoxin in rats. Administration of 5 mg/kg of the glucocorticoid receptor antagonist RU 38486 2 hours before endotoxin injection did not affect the inhibition of amino acid uptake. In vitro addition of plasma from endotoxemic rats to incubated rat soleus muscles inhibited amino acid uptake by approximately 30%. This effect of endotoxic plasma also was noted when muscles were from rats that had been treated with RU 38486 and when RU 38486 was present in the incubation medium. Results confirm previous reports of reduced muscle amino acid transport during endotoxemia and of the presence of a circulating factor that inhibits muscle amino acid uptake in this condition. Data suggest that inhibited muscle amino acid transport during endotoxemia is not regulated by glucocorticoids. PMID- 8412775 TI - Insulin sensitivity of splanchnic and peripheral adipose tissue in vivo in morbidly obese man. AB - Epidemiologic studies demonstrate an association between increased waist to hip ratio ([WHR] android obesity, central obesity) and diabetes mellitus in man. To study the relative insulin sensitivity of splanchnic versus peripheral adipose tissue, portal vein catheterization via the collapsed umbilical vein was performed in 14 morbidly obese subjects at the time of surgery. Catheters were also placed in a peripheral artery and antecubital vein such that simultaneous arterio-venous (A-V) differences (glycerol, free fatty acids [FFA], and lactate) could be determined. After two baseline samples obtained 3 minutes apart, 25 g intravenous (i.v.) glucose (14 subjects) was administered over a 2-minute period, with samples being obtained every 5 minutes for 30 additional minutes. Arterial plasma glycerol levels decreased from 173.9 +/- 17.4 mumol/L at baseline to 89.1 +/- 7.6 mumol/L at 30 minutes (P < .01). Peripheral and splanchnic A-V glycerol differences were similar at baseline, but within 10 minutes after glucose administration the difference across the splanchnic area decreased by 52% and remained significantly less than that across the periphery (P < .01). Despite a 49% decrease in arterial plasma glycerol level, plasma FFA level decreased only 18.3% over the 30-minute period (942 +/- 74.8 to 770.0 +/- 76 mumol/L, NS). These studies in morbidly obese man (glycerol data) indicate a greater insulin sensitivity of splanchnic adipose tissue than of peripheral adipose tissue. Thus hypertrophy of fat in the splanchnic area might be an expected consequence of the hyperinsulinemia associated with insulin-resistant states. PMID- 8412776 TI - Energy metabolism in response to overfeeding in young adult men. AB - The effect of overfeeding on energy metabolism was evaluated in four young adult men of normal weight. Subjects were fed a hypocaloric diet for 8 days followed by an 8-day hypercaloric diet to maximize differences in the metabolic response to the two diets. Energy intake during the hypocaloric and hypercaloric phases was 1.32 and 2.26 times resting metabolic rate (RMR), respectively. Additional calories during hypercaloric feeding were provided as 1,659 +/- 40 kcal/d carbohydrate and lipid. Total daily energy expenditure (TDEE) was measured by the doubly labeled water technique and RMR by indirect calorimetry. Nonresting energy expenditure (NREE) was calculated as the difference between TDEE and RMR. During the 8-day hypocaloric diet period, energy intake was approximately 340 kcal/d less than the measured rate of TDEE (2,384 +/- 219 kcal/d) and the subjects lost 0.8 +/- 0.3 kg body weight. During the 8-day hypercaloric diet period, TDEE increased by 425 +/- 103 kcal/d (P = .03), which dissipated 25% +/- 5% of the increased energy intake. One fourth of the increase in TDEE was caused by an increase in RMR (110 +/- 24 kcal/d, P = .02) and the remainder by an increase in NREE (316 +/- 103 kcal/d, P = .05). These results demonstrate that only a small portion of excess energy intake is dissipated by an increase in energy expenditure. NREE can be an important component of the increase in energy metabolism during overfeeding. PMID- 8412777 TI - Refeeding after fasting increases apparent oxidation of N-3 and N-6 fatty acids in pregnant rats. AB - The intake, excretion, and accumulation of long-chain fatty acids was analyzed to test the hypothesis that during pregnancy in the rat whole-body partitioning of n 6 and n-3 fatty acids between net accumulation or disappearance (apparent oxidation) is determined by both maternal energy status and fetal development. From midpregnancy to term in ad libitum-fed rats consuming-rodent chow, 30% more saturates and monounsaturates accumulated in the whole body than were consumed, whereas 28% of dietary n-6 fatty acids and 55% of dietary n-3 fatty acids were apparently oxidized. After 48 hours of fasting during midpregnancy (days 13 to 15) followed by refeeding to term, net accumulation of saturates and monounsaturates was equivalent to intake, but whole-body disappearance of n-6 fatty acids exceeded intake by 6% (NS), whereas n-3 fatty acid disappearance exceeded intake by 43%. Thus during refeeding after fasting, dietary n-6 and n-3 fatty acids were apparently completely oxidized and there was actually a net loss of n-3 fatty acids from whole-body stores. Fasting during midpregnancy did not significantly affect fetal weight gain or maternal gestational hyperlipidemia toward term. We conclude that during pregnancy n-6 and n-3 fatty acids are not only required for maternal and fetal structural and storage lipids, but are also used to meet energy requirements, especially during refeeding after fasting. PMID- 8412778 TI - Effect of ethanol ingestion on nucleotides and glycolytic intermediates in erythrocytes and purine bases in plasma and urine: acetaldehyde-induced erythrocyte purine degradation. AB - The effect of ethanol on nucleotides and glycolytic intermediates in erythrocytes and purine bases in plasma and urine was investigated. Ethanol ingestion (0.45 mL/kg body weight) increased plasma concentrations and urinary excretion of oxypurines (hypoxanthine and xanthine) and concentrations of adenosine monophosphate (AMP), adenosine diphosphate (ADP), and glyceraldehyde 3 phosphate+dihydroxyacetonephosphate in erythrocytes. In an in vitro incubation study using erythrocytes, acetaldehyde increased the concentrations of AMP, ADP, and glyceraldehyde 3-phosphate+dihydroxyacetonephosphate in erythrocytes as well as the concentration of hypoxanthine in the incubation medium. These results suggest that acetaldehyde (a metabolite of ethanol) induces an increase in purine degradation by erythrocytes and then contributes to the ethanol-induced enhanced purine degradation in vivo. PMID- 8412779 TI - The impact of metformin therapy on hepatic glucose production and skeletal muscle glycogen synthase activity in overweight type II diabetic patients. AB - The effect of metformin therapy on glucose metabolism was examined in eight overweight newly presenting untreated type II diabetic patients (five males, three females). Patients were treated for 12 weeks with either metformin (850 mg x 3) or matching placebo using a double-blind crossover study design; patients were studied at presentation and at the end of each treatment period. Insulin action was assessed by measuring activation of skeletal muscle glycogen synthase (GS) before and during a 4-hour hyperinsulinemic euglycemic clamp (100 mU.kg-1 x h-1). Metformin therapy was associated with a significant decrease in fasting blood glucose (6.8 +/- 0.6 v 8.3 +/- 0.9 mmol.L-1, P < .01) and glycosylated hemoglobin ([HbA1] 7.7% +/- 0.4% v 8.5% +/- 0.5%, P < .01) levels. Fasting hepatic glucose production (HGP) was also significantly decreased following metformin therapy (1.98 +/- 0.13 v 2.41 +/- 0.20 mg.kg-1 x min-1, P < .02), whereas fasting insulin and C-peptide concentrations remained unaltered. The decrease in basal HGP correlated closely with the decrease in fasting blood glucose concentration (r = .92, P < .001). Insulin-stimulated glucose uptake was assessed using the hyperinsulinemic euglycemic clamp technique and was increased post-metformin (3.8 +/- 0.6 v 3.1 +/- 0.7 mg.kg-1 x min-1, P < .05). This was primarily the result of increased nonoxidative glucose metabolism (1.1 +/- 0.6 v 0.4 +/- 0.6 mg.kg-1 x min-1, P < .05); oxidative glucose metabolism did not change. Metformin had no measurable effect on insulin activation of skeletal muscle GS, the rate-limiting enzyme controlling muscle glucose storage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412780 TI - Growth hormone acutely stimulates skeletal muscle but not whole-body protein synthesis in humans. AB - In a previous study, a 6-hour local infusion of growth hormone (GH) into the brachial artery of normal subjects stimulated net muscle protein anabolism by augmenting skeletal muscle protein synthesis. In the present study, we examined whether systemically infused GH affects forearm and whole-body protein metabolism. Normal volunteers aged 18 to 24 years (n = 8) were given an 8-hour systemic infusion of 3H-phenylalanine and 14C-leucine. Between 90 and 120 minutes of tracer infusion, basal samples for determination of forearm and whole-body amino acid kinetics were taken. GH was then infused at 0.06 micrograms/kg/min, increasing GH concentration from 2.4 +/- 0.3 to 32 +/- 3 ng/mL. Systemic insulin like growth factor 1 (IGF-1) level increased from 224 +/- 20 to 262 +/- 21 ng/mL (basal v 6-hour, P < .01). By 6 hours, GH suppressed forearm phenylalanine and leucine net release (each P < .05) by increasing 3H-phenylalanine (66%, P < .05) and 14C-leucine (13%, P < .05) extraction or disposal (Rd). Whole-body leucine rate of appearance ([Ra] an index of whole-body proteolysis) and nonoxidative leucine Rd (whole-body protein synthesis) did not change over the course of the GH infusion, whereas oxidative leucine Rd decreased (20%, P < .03). Acute stimulation of muscle but not whole-body protein synthesis by systemically infused GH suggests that muscle protein is acutely and specifically regulated by GH. PMID- 8412781 TI - Biobehavioral mechanisms in lipid metabolism and atherosclerosis: an overview. PMID- 8412782 TI - Sex steroids, psychosocial factors, and lipid metabolism. PMID- 8412783 TI - Insulin, health behaviors, and lipid metabolism. PMID- 8412784 TI - Stress and lipoprotein metabolism: modulators and mechanisms. PMID- 8412785 TI - Lipids, growth, and development. PMID- 8412786 TI - Low or lowered cholesterol and risk of death from suicide and trauma. PMID- 8412787 TI - Introduction: blood coagulation. PMID- 8412788 TI - Proteolytic enzymes in coagulation, fibrinolysis, and complement activation. Part A. Mammalian blood coagulation factors and inhibitors. PMID- 8412789 TI - Modular design of proteases of coagulation, fibrinolysis, and complement activation: implications for protein engineering and structure-function studies. PMID- 8412790 TI - Factor VIII and factor VIIIa. PMID- 8412791 TI - Characterization of factor IX defects in hemophilia B patients. PMID- 8412792 TI - Characterization of dysfunctional factor VIII molecules. AB - Immunopurification and characterization of dysfunctional factor VIII-like molecules in CRM-positive and CRM-reduced hemophilia A permit correlation of structural changes with molecular defects. The technique described here is sufficiently sensitive to characterize the molecular mass and enzymatic fragments of the factor VIII chains in patients with as little VIII: Ag as 0.05 units/ml. Specific abnormalities have been identified in 5 of the first 24 samples tested. In each case, the mutation responsible for factor VIII dysfunction has been determined by sequencing a part of the abnormal gene. Mutations have been identified that abolish critical thrombin cleavage sites or which generate new N glycosylation sites. The technique provides a useful approach to the study factor VIII structure-function relationships, and it has the potential to clarify further the molecular basis of factor VIII procoagulant activity. PMID- 8412793 TI - Extrinsic pathway proteolytic activity. PMID- 8412794 TI - Tissue factor pathway inhibitor. PMID- 8412795 TI - Mutational analysis of receptor and cofactor function of tissue factor. PMID- 8412796 TI - Factor V. PMID- 8412797 TI - Site-directed mutagenesis and expression of coagulation factors VIII and V in mammalian cells. PMID- 8412798 TI - Assembly of prothrombinase complex. AB - The approaches described in this article have resulted in an increased understanding of the reaction steps involved in the stabilization and assembly of the prothrombinase complex. Because prothrombinase is considered an archetype for some of the other coagulation complexes, the quantitative information derived from these studies (Table I) provides the framework for future studies of prothrombinase and suggests experimental approaches for studies of the other analogous coagulation reactions. PMID- 8412799 TI - Procoagulant activities expressed by peripheral blood mononuclear cells. AB - These combined data support the concept that the procoagulant response elicited by mononuclear cells, particularly monocytes, is accomplished through regulated binding site-mediated (or perhaps "receptor"-mediated) assembly of proteolytic activities at their membrane surface. Because the work of several laboratories indicate that the monocytes provide the appropriate membrane surface for the assembly and function of all the coagulation complexes required for thrombin production in vivo, monocytes may provide a unique opportunity to investigate how coagulant reactions are regulated on cell surfaces through both receptor-mediated events as well as by channeling a product of one reaction to serve as a mediator of a second reaction. PMID- 8412800 TI - Meizothrombin: active intermediate formed during prothrombinase-catalyzed activation of prothrombin. PMID- 8412801 TI - Identification and characterization of mutant thrombins. PMID- 8412802 TI - Synthetic selective inhibitors of thrombin. PMID- 8412803 TI - Quantifying thrombin-catalyzed release of fibrinopeptides from fibrinogen using high-performance liquid chromatography. PMID- 8412804 TI - Protein C activation. PMID- 8412805 TI - Molecular approach to structure-function relationship of human coagulation factor XIII. PMID- 8412806 TI - Protein C inhibitor. PMID- 8412807 TI - Immunochemical techniques for studying coagulation proteins. PMID- 8412808 TI - Isolation of intact modules from noncatalytic parts of vitamin K-dependent coagulation factors IX and X and protein C. PMID- 8412809 TI - Role of propeptide in vitamin K-dependent gamma-carboxylation. PMID- 8412810 TI - Expression of recombinant vitamin K-dependent proteins in mammalian cells: factors IX and VII. PMID- 8412811 TI - Thioester peptide chloromethyl ketones: reagents for active site-selective labeling of serine proteinases with spectroscopic probes. PMID- 8412812 TI - Peptide chloromethyl ketones as labeling reagents. PMID- 8412813 TI - Factor XII: Hageman factor. PMID- 8412814 TI - Active site-specific assays for enzymes of coagulation and fibrinolytic pathways. PMID- 8412815 TI - Kinetic characterization of heparin-catalyzed and uncatalyzed inhibition of blood coagulation proteinases by antithrombin. PMID- 8412816 TI - Factor XI: structure-function relationships utilizing monoclonal antibodies protein modification, computational chemistry, and rational synthetic peptide design. PMID- 8412817 TI - Human factor IX and factor IXa. PMID- 8412819 TI - Modelling domain-knowledge: a step toward intelligent data management. PMID- 8412818 TI - Intelligent processing of loosely structured documents as a strategy for organizing electronic health care records. AB - Loosely structured documents can capture more relevant information about medical events than is possible using today's popular databases. In order to realize the full potential of this increased information content, techniques will be required that go beyond the static mapping of stored data into a single, rigid data model. Through intelligent processing, loosely structured documents can become a rich source of detailed data about actual events that can support the wide variety of applications needed to run a health-care organization, document medical care or conduct research. Abstraction and indirection are the means by which dynamic data models and intelligent processing are introduced into database systems. A system designed around loosely structured documents can evolve gracefully while preserving the integrity of the stored data. The ability to identify and locate the information contained within documents offers new opportunities to exchange data that can replace more rigid standards of data interchange. PMID- 8412820 TI - Marking up is not enough. PMID- 8412821 TI - The explanatory role of events in causal and temporal reasoning in medicine. AB - The logic of time and the way we reason about time is intrinsically connected with the way we reason about causality. In this paper, we focus our attention on some of the less obvious ways in which reasoning about time and causality interact. It is explained why in temporal reasoning a firm distinction has to be made between the ontology, i.e., what happens, and the way we describe the ontology. Temporal events need to be redescribed in such a way that they causally explain why some of the events are followed by the others. While building a temporal/causal theory, certain events may be omitted, not because they do not play a causal role, but because they do not play an explanatory role. In doing so, it is possible to eliminate the distinction between theories representing time as dense, and theories that represent time as discrete. PMID- 8412822 TI - Documenting an event and its evolving meaning. PMID- 8412823 TI - The Unified Medical Language System. AB - In 1986, the National Library of Medicine began a long-term research and development project to build the Unified Medical Language System (UMLS). The purpose of the UMLS is to improve the ability of computer programs to "understand" the biomedical meaning in user inquiries and to use this understanding to retrieve and integrate relevant machine-readable information for users. Underlying the UMLS effort is the assumption that timely access to accurate and up-to-date information will improve decision making and ultimately the quality of patient care and research. The development of the UMLS is a distributed national experiment with a strong element of international collaboration. The general strategy is to develop UMLS components through a series of successive approximations of the capabilities ultimately desired. Three experimental Knowledge Sources, the Metathesaurus, the Semantic Network, and the Information Sources Map have been developed and are distributed annually to interested researchers, many of whom have tested and evaluated them in a range of applications. The UMLS project and current developments in high-speed, high capacity international networks are converging in ways that have great potential for enhancing access to biomedical information. PMID- 8412824 TI - A new architecture for integration of heterogeneous software components. AB - An architecture is described that integrates existing applications in a network wide system. The architecture follows the new open software paradigm, and defines kernel and application services that collaboratively solve the tasks of end-users and provide them with an intuitive user-interface. This paper describes the message language and the kernel mechanism for addressing application services. The architecture has been developed as much as possible to conform with current standards. PMID- 8412825 TI - Forecasting survival in the medical intensive care unit: a comparison of clinical prognoses with formal estimates. AB - Physicians often need to make prognostic judgments. In the present study, the accuracy was explored of survival estimates for patients in the Medical Intensive Care Unit (MICU). Estimates were made by physicians and nurses several times during each patient's stay in the MICU and were compared to those of the APACHE II scale, a widely used quantitative index for critically ill patients. ROC curve and calibration curve analyses were performed to assess the accuracy of these estimates. Results revealed that MICU personnel were fairly accurate discriminators of patients who survived vs. who died, although there was a consistent tendency to underestimate survival. In addition, there was some relationship between the level of physician training and forecasting accuracy, but only within the patient's first 24 hours in the MICU. Finally, the estimates of physicians did not differ significantly from those of the APACHE II scale. Physicians tended to be better calibrated in their predictions, while the APACHE II scale was slightly superior in terms of discrimination. PMID- 8412826 TI - Computer-assisted instruction in probabilistic reasoning during the inpatient medicine clerkship. AB - The acceptability and utility of computer-assisted instruction in probabilistic reasoning was assessed for medicine clerkship students. After a pretest, the experimental (n = 40), but not the control students (n = 39), completed a program that we designed. The program contained the test and its answers. After program exposure, experimental students rated their knowledge of the program's content significantly higher (p = 10(-4)) than control students. On the identical posttest, experimental students also scored significantly higher than control students (p = 10(-4)) and improved their scores significantly more (p = 10(-3)). They rated ease-of-use items significantly higher than content-relevance items (p = 10(-4)). We conclude that computer-assisted instruction in probabilistic reasoning is acceptable to clerkship students, and that it may improve their knowledge and skills in this area. However, students may rate the vehicle of this instruction more highly than its content. PMID- 8412827 TI - Stratified initial values and change scores. AB - A common question that is encountered in many studies is relating a change in a variable to the variable's initial value. It is well known that regressing the change on the initial value creates a spurious association and misleads the unwary. One attempt to cope with this problem has been to stratify that data based on the initial value. We show that this approach does not solve the problem. Theoretical and simulation results for independent normal and independent exponential random variables are provided. PMID- 8412828 TI - A methodology for determining patients' eligibility for clinical trials. AB - The task of determining patients' eligibility for clinical trials is knowledge and data intensive. In this paper, we present a model for the task of eligibility determination, and describe how a computer system can assist clinical researchers in performing that task. Qualitative and probabilistic approaches to computing and summarizing the eligibility status of potentially eligible patients are described. The two approaches are compared, and a synthesis that draws on the strengths of each approach is proposed. The result of applying these techniques to a database of HIV-positive patient cases suggests that computer programs such as the one described can increase the accrual rate of eligible patients into clinical trials. These methods may also be applied to the task of determining from electronic patient records whether practice guidelines apply in particular clinical situations. PMID- 8412829 TI - Reconstructing medical problem solving competence: MACCORD. AB - The building of medical knowledge-based systems involves the reconstruction of methodological principles and structures within the various subdomains of medicine. ACCORD is a general methodology of knowledge-based systems, and MACCORD its application to medicine. MACCORD represents the problem solving behavior of the medical expert in terms of various types of medical reasoning and at various levels of abstraction. With MACCORD the epistemic and cognitive processes in clinical medicine can be described in formal terminology, covering the entire diversity of medical reasoning. MACCORD is close enough to formalization to make a significant contribution to the fields of medical knowledge acquisition, medical didactics and the analysis and application of medical problem solving methods. PMID- 8412830 TI - Semantics from loosely structured electronic health records. PMID- 8412831 TI - Midwifery legalised in British Columbia. PMID- 8412832 TI - Safe Motherhood--the first decade. AB - In 1987 an international project, the Safe Motherhood Initiative, was commenced with the aim of reducing, by half by the Year 2000, the 500,000 maternal deaths which occur each year throughout the world. In this paper the progress of the Initiative is described and reviewed. The crucial role of the midwife in reducing maternal mortality is recognised and the work to be done in the future is identified. This paper was given as a keynote address at the 23rd Congress of the International Confederation of Midwives in Vancouver in May, 1993. PMID- 8412833 TI - Testing for fetal abnormality in routine antenatal care. AB - The detection of fetal abnormality is a major component of routine antenatal care. A variety of techniques are now in use, although these are constantly being modified in the pursuit of more accurate and earlier detection. In this paper we draw attention to the distinction between screening and diagnostic tests, and describe the techniques which have been most commonly used in the UK: serum screening for neural tube defects; screening for Down's syndrome; ultrasound scanning; amniocentesis and chorionic villus sampling. PMID- 8412834 TI - Perceptions of pain and pain relief in labour: the gulf between experience and observation. AB - In a national sample of 10,702 women delivered in one week in 1990 data on their perceptions of pain and the effectiveness of pain relief methods were compared with those of the professionals attending them. Professionals' conceptions of pain relief tended to be restricted to pharmacological methods. Overall, the level of agreement about effectiveness of pain relief was quite low, with medical staff commonly believing they were providing adequate pain relief while the women reported it as unsatisfactory. This was particularly the case for pethidine, while 'Entonox' was rated more highly by women than staff. Professionals were significantly more likely to agree with one another about effectiveness than to agree with the women, and were less likely to have responded where the women judged their pain relief as poor. PMID- 8412835 TI - Satisfaction with postnatal care--the choice of home or hospital. AB - This paper reports the findings of a study of client satisfaction with postnatal midwifery care. Women could choose one of two forms of care; either domiciliary care following early discharge, or hospital care until discharge. Consumers' perceptions of their postnatal care were examined at the end of the period of care. Women assessed the midwives' interest and caring, education and information provided, their own progress with feeding and baby care, and their own physical and emotional health. They were also asked about their expectations of and gains from postnatal care. The findings indicated that women choosing domiciliary care and women choosing hospital care had different expectations of their postnatal care, but were largely satisfied with the quality of the care they chose. The women who chose domiciliary care rated their postnatal care more highly than the women who stayed in hospital. The findings reinforce the importance of providing women with choices for the maternity care which best suits their needs. PMID- 8412836 TI - An evaluation of postnatal care individualised to the needs of the woman. AB - Traditionally, in Scotland women have been visited daily in their own home until the 10th postnatal day. There has been concern about the ability of such a service to offer continuity of care. A 'before and after' study was undertaken at Glasgow Royal Maternity Hospital to assess the effects of introducing individualised postnatal care. In the first stage information was collected on 106 women to obtain baseline information. In the second stage information was collected on 114 women who had experienced individualised care. The women in the two stages were not significantly different in terms of age, parity, mode of delivery and problems encountered. The average number of midwives visiting during the postnatal period fell from 3.7 to 2.5 and the average number of visits fell from 6.5 to 5.7 visits. Both forms of care, standard and individualised, were popular with women, but the proportion who felt a daily visit necessary dropped significantly. Continuity of care was improved with individualising care to the needs of the woman and women were satisfied with this type of care. The change is likely to be cost effective, since a higher quality of care can be provided for the same, or slightly less cost. PMID- 8412837 TI - Health after childbirth: a comparison of normal and assisted vaginal delivery. AB - 413 women were included in the study: 100 who had had a normal delivery and 313 who had participated in a randomised controlled comparison of forceps or ventouse delivery. The study comprised a questionnaire and assessment at 24-48 hours after delivery and a questionnaire in the second year after delivery. There was significantly more morbidity in the women in the instrumental delivery group compared to women in the normal delivery group at both the early assessment and long-term follow-up. PMID- 8412838 TI - Position/policy statements adopted by ICM council at meeting in Vancouver--May 1993. PMID- 8412839 TI - Passage of the menopause is followed by haemostatic changes. AB - The passage of the menopause has been reported to be followed by a steadily increasing risk of cardiovascular disease (CVD). Changes in the concentrations of certain coagulation factors and fibrinolytic components are considered risk factors for CVD. We evaluated the differences in some of these variables between a premenopausal group (A) (n = 28) and two postmenopausal groups, one of women less than 18 months past the menopause (B) (n = 28), the other of women more than 18 months past the menopause (C) (n = 21). The variables measured were serum oestradiol content, plasma antithrombin III (AT III) activity, protein C activity and the plasma concentrations of tissue type plasminogen activator (t-PA) antigen, plasminogen activator inhibitor (PAI-1) antigen, and fibrinogen. As compared with the premenopausal women (group A), group C showed significantly higher values for AT III and protein C activity and for t-PA and PAI-1 antigen; and group B and C both showed significantly higher fibrinogen concentrations. This probably means that haemostatic balance was maintained in the postmenopausal women, although the increased concentrations of fibrinogen and PAI-1 might constitute risk factors for the development of cardiovascular disease. PMID- 8412840 TI - Blood serotonin levels in postmenopausal women: effects of age and serum oestradiol levels. AB - This study investigated the effect of ageing and reduction in ovarian function on whole blood serotonin levels and the effect of the oestrogen replacement in postmenopausal women on blood serotonin levels. Amenorrheic, natural postmenopausal and ovariectomised women had lower blood serotonin levels than regularly menstruating women. Blood serotonin levels increased after oestrogen replacement in postmenopausal women to values similar to those observed in regularly menstruating women. In ovariectomised women, the blood serotonin levels were inversely correlated with age (P < 0.01). In women with different serum oestradiol levels ranging 30-1335 pg/ml, a direct relationship was observed between serum oestradiol levels and whole blood serotonin levels (P < 0.001). It was concluded that whole blood serotonin concentration is reduced during menopause by suppression of ovarian function but may also be an effect of ageing. PMID- 8412841 TI - Characteristics of headache at menopause: a clinico-epidemiologic study. AB - The prevalence and characteristics of primary headaches in a large sample of postmenopausal women were investigated. Seventy-six out of 556 women (13.7%) were affected by headache of either the migraine or tension type. In 82% of cases onset had preceded the menopause. The postmenopausal course of headaches with a premenopausal onset differed according to type of headache and type of menopause. Indeed, while migraine improved in almost two-thirds of cases, tension-type headache worsened or did not change in 70% of cases. However, in women who had undergone surgical ovariectomy, the natural course of migraine was worse than in those who had a physiological menopause (P = 0.003). Among the symptoms covered by the Kuppermann Index, only anxiety and insomnia were correlated with headache. The favourable course of migraine in the postmenopausal period can be attributed primarily to the absence of variations in sex hormone levels although psychological factors also seem to play a fundamental role. PMID- 8412842 TI - Reproductive factors as predictors of bone density and fractures in women at the age of 70. AB - The importance of reproductive and other health and social factors as predictors of bone mineral density (BMD) and fractures was investigated in a longitudinal study of 70-year-old women. At 70 years of age there was no correlation between menarcheal age, menopausal age, duration of fertile period and parity and BMD or a history of fractures. Bilateral oophorectomy (mean age at surgery 45.3 +/- 3.6 years) and the duration of tobacco smoking were negatively correlated to BMD at 70 years of age. Tobacco smoking was positively correlated to a history of fractures and body mass index (BMI) was negatively correlated to a history of fractures at 70 years of age. In a stepwise multiple regression model, height, body mass index and a history of hysterectomy were found to have a contributory, positively-correlated independent explanatory value for BMD at 70 years. In a logistic multiple regression analysis, bilateral oophorectomy was the only independent explanatory factor for fractures at 70 years of age. BMD was lower in women who had undergone bilateral oophorectomy compared with controls at 70 years of age. Hysterectomized women without bilateral oophorectomy showed a higher BMD in the right calcaneus at 70 years of age compared with controls. When comparing the reduction in bone mineral density between 70 and 76 years of age, the fifth quintile (women with the highest BMD) at 70 years of age decreased 2.7 times more (P < 0.01) than the first quintile. A longitudinal examination between 70 and 76 years revealed that BMD at the age of 70 was a significant (P < 0.01) predictor for fractures occurring between 70 and 76 years of age. PMID- 8412843 TI - Effects of oestrogen on hypothalamic beta-endorphin in ovariectomized and old rats. AB - Immunocytochemical beta-endorphin (beta-EP) staining and ultrastructural observations in the hypothalamus were compared in normal mature female rats, ovariectomized rats, and aged female rats. The effects of oestradiol benzoate (EB) on the hypothalamus were studied. The female Wistar rats were divided into seven groups, as follows: 40-day-old rats (Mature), 54-day-old rats ovariectomized at 40 days (Ovx), rats ovariectomized at 40 days and injected with 0.1 mg EB daily for 7 days (Ovx + e) rats ovariectomized and injected with one dose of 1 mg EB (Ovx + E), 500-day-old (Old), old rats injected with 0.1 mg EB daily for 7 days (Old + e), and old rats injected with 1 mg EB (Old + E). In the Ovx and Old groups, beta-EP-positive cells in the arcuate nucleus were rarely seen, as compared with the Mature group. The staining of beta-EP-positive cells in Ovx + e was slightly recovered and that in Ovx + E was almost completely recovered. However, no recovery of beta-EP-positive cells was seen in the Old + e or Old + E groups. The number of nerve fibers in the median eminence were reduced in both the Ovx and Old groups, as compared with the Mature group. There was no relationship between changes in these numbers and oestrogen replacement in the Old group, but in the Ovx group oestrogen replacement brought about recovery of these numbers. The number of glial cells increased after oestrogen replacement in both the Ovx and Old groups. The frequency of giant mitochondria in the neurons in the arcuate nucleus decreased after oestrogen replacement in the Old group. PMID- 8412844 TI - Hormonal treatments modulate pulsatile plasma growth hormone, gonadotrophin and osteocalcin levels in postmenopausal women. AB - Oestrogen plays a role in modulating growth hormone (GH), luteinizing hormone (LH), follicle-stimulating hormone (FSH) and osteocalcin secretion in women. Indeed, the postmenopausal period is characterized by changes in plasma GH, LH, FSH and osteocalcin levels. The aim of the present study was to investigate the changes in the secretory patterns of these hormones in postmenopausal women under different therapeutic regimens. A total of 20 subjects took part in the study. They were subdivided into four groups comprising 5 untreated postmenopausal women (time since menopause 1-5 years), 5 postmenopausal women receiving steroid hormone replacement therapy, 5 postmenopausal women receiving salmon calcitonin and 5 fertile women with regular menstrual cycles. Blood samples were collected every 15 min for 4 h and hormone levels were measured by radioimmunoassay. Plasma GH levels fell in the postmenopausal women, but were restored under oestrogen progestogen treatment. The pulsatile patterns of plasma LH and FSH did not show significant differences in the treated and untreated women. Plasma osteocalcin levels showed episodic fluctuations and mean levels decreased under steroid hormone or calcitonin treatment. No significant correlation was observed between plasma GH and osteocalcin or gonadotrophin levels or body mass index. PMID- 8412845 TI - The menopausal age and climacteric complaints in Thai women in Bangkok. AB - Age at menopause, associated factors and related clinical symptoms were recorded in a sample of Thai women living in the Bangkok area. Interviews by trained nurses were offered to a sample of 2375 women aged 45-59 years selected at random in 19 of the 54 Bangkok Metropolis Administration health centers. Full reports were obtained from 2354 women. Fifty-six percent of the study population were classified as postmenopausal, i.e. having no vaginal bleeding during the last 12 months, 12% were perimenopausal, i.e. having irregular vaginal bleeding during the last 12 months and 31% were premenopausal, i.e. having regular vaginal bleeding during the last 12 months. The average age at menopause was 49.5 +/- 3.6 years and was not related to body weight, height, age at menarche, level of education, smoking or use of oral contraceptives. However, high parity was significantly correlated to delayed menopause. Clinical symptoms of oestrogen deficiency were reported at a significantly higher rate in the perimenopause group, but the premenopause and postmenopause groups did not differ in their complaints about any symptom, including hot flushes. The most striking effect of menopause was a dramatic loss of sexual desire in 86.9% of postmenopausal women. However, the situation did not induce any specific complaint or any request for medical support. PMID- 8412846 TI - Detection of bacteriocins produced by Lactobacillus plantarum strains isolated from different foods. AB - Bacterial strains (106 in toto) isolated from different foods and identified as Lactobacillus plantarum were screened for antagonistic activities against other bacteria under conditions which eliminated the effects of organic acids and hydrogen peroxide. Five isolates were shown to be bacteriocin producers, and the bacteriocins, on the basis of their host range inhibition and cross inhibition were all different. The bacteriocins were preliminarily characterized by temperature stability, sensitivity to proteolytic, lipolytic and glycolytic enzymes and precipitation with ammonium sulphate. PMID- 8412847 TI - A simple DNA polymerase chain reaction method to locate and define orientation of specific sequences in cloned bacterial genomic fragments. AB - A simple DNA polymerase chain reaction (PCR) method, to rapidly locate and define the orientation of a particular sequence within a cloned bacterial genomic fragment several kilobases (kb) long, is described. The technique is particularly useful when cloning (by DNA PCR amplification) a specific sequence of a conserved gene from several microorganisms following an homology probing approach. The method requires two universal primers derived from the vector, two specific primers derived from each end of the specific sequence in inverted tail-to-tail directions, and a single round of PCR. In addition, PCR conditions applicable to DNA inserts having a G + C content up to 75% (e.g. Pseudomonas and actinomycete genomic fragments), and allowing efficient amplification of DNA fragments up to 7 kb long, are described and discussed. PMID- 8412848 TI - Influence of fermentation conditions on specific activity of the enzymes alcohol and aldehyde dehydrogenase from yeasts. AB - The effects of anaerobic, semi-aerobic and short aeration fermentation conditions and the addition of ergosterol and oleic acid to musts on the specific activity of alcohol and aldehyde dehydrogenase (ADH and ALDH) from two yeast species, Saccharomyces cerevisiae and Torulaspora delbrueckii, were studied. ADH I biosynthesis only occurred during the first few hours of fermentation. ADH II from S. cerevisiae and ALDH-NADP+ from the two yeast species behaved as constitutive enzymes under all fermentation conditions. ADH II from T. delbrueckii was only synthesized in small amounts, and its activity was always lower than in S. cerevisiae, where it was responsible for the termination of alcoholic fermentation during the steady growth phase. PMID- 8412849 TI - Contributions of oxygen dissociation and convection to the behavior of a compartmental oxygen transport model. AB - We previously derived a compartmental model for oxygen transport in vascular blood in which lumped terms, representing convective and oxygen dissociative effects, were obtained from a distributed model by space averaging. In this paper, we compare the results of this model and those of another compartmental model from the literature in which these terms were selected arbitrarily. The use of space-averaged parameters resulted in a more distinct difference in calculated partial pressures of oxygen between capillary and venule compartments, more accurate distribution patterns for partial pressures of oxygen through the venule compartments, and a capability to simulate conditions under which mean oxygen partial pressure in tissue is higher than in the venules. These results are supported by available experimental findings. Results also showed that the employment of space-averaged convective terms had a greater effect on the distribution of compartmental partial pressures than did the use of the cross sectional averaged oxygen-hemoglobin binding function. The latter produced significant changes only for certain compartments, and only under extreme physiologic conditions. The results demonstrate our model's ability to reflect relationships among capillary, venule, and tissue compartmental partial pressures under varying conditions. PMID- 8412850 TI - Vasomotion in the totally denervated transplanted rabbit ear chamber. PMID- 8412851 TI - Intravital and electron microscopic observation of Ito cells in rat hepatic microcirculation. AB - Intralobular distribution of Ito cells (fat-storing cells) in hepatic microcirculatory units was investigated through intravital fluorescence microscopy. By using a low-level ultraviolet epi-illumination and a sensitive silicon intensified target camera, the intrahepatic autofluorescence was visualized and digitally processed. In rats fed an ordinary diet, the ultraviolet excited autofluorescence was composed of at least two different origins that could not be spectrophotometrically distinguished, namely, multiple patchy autofluorescence activities along the sinusoids and terminal hepatic venules and diffuse parenchymal autofluorescence. Multiple patchy activities showed a rapid photobleaching phenomenon under the continuous ultraviolet excitation. These fluorescent activities were completely eliminated by depleting the intrahepatic retinoid contents using a vitamin A-deficient diet for 4 weeks. Furthermore repeated administration of vitamin A significantly enhanced the patchy fluorescent activities. Electron microscopy revealed that these vitamin A fluorescent activities are colocalized with the fat droplets in Ito cells, providing evidence that Ito cells exist not only in perisinusoidal spaces but also in the perivascular spaces of the terminal hepatic and portal venules. The current intravital technique thus provides a new method to observe Ito cells in hepatic microcirculatory units in vivo. PMID- 8412852 TI - Effect of erythrocyte deformability on in vivo red cell transit time and hematocrit and their correlation with in vitro filterability. AB - Indicator dilution techniques were applied to measure mean transit time of fluorescently labeled red blood cells (RBCs) (TTRBC) and plasma (TTpl) between functionally paired arterioles and venules (A-V) in cremaster muscle (rat) for normal RBCs and cells hardened by in vitro incubation in graded concentrations of glutaraldehyde. Dispersion of a bolus introduced into the contralateral femoral artery permitted computation of TT by cross-correlation of fluorescence intensity time curves in A-V pairs. Parallel in vitro assessments of RBC deformability were made by filtration through 5-microns pore Nuclepore filters to express deformability in terms of the ratio of resistance to flow through a pore with RBCs present to that with suspending medium alone, beta. The average microvascular hematocrit (Hmicro) normalized with respect to systemic hematocrit (Hsys) was calculated from TTRBC and TTpl. For 26 A-V pairs of the third and fourth orders of branching, TTRBC averaged 0.63 sec for normal control cells (beta = 2.61), and TTpl averaged 0.85 sec with an average TTRBC/TTpl equal to 0.85. The corresponding value of Hmicro/Hsys was significantly < 1 and averaged 0.87. This greater value of Hmicro/Hsys compared to direct measurements in the literature was attributed to the unique ability of the indicator dilution technique to account for red cell flux throughout the network. For hardened RBCs with beta < 10, TTRBC/TTpl and Hmicro/Hsys increased on average 30%, but were weakly correlated with increasing beta due to redistribution of RBCs throughout pathways of lesser resistance. However, as beta rose from 10 to 20, these pathways became overwhelmed by hardened RBCs and TTRBC/TTpl increased threefold due to retardation of the RBC flux, with a concomitant rise in Hmicro/Hsys. These results clearly demonstrate the extent to which diminished RBC deformability of a magnitude found in clinical disorders may affect microvascular perfusion. PMID- 8412853 TI - Changes in vessel ultrastructure during ischemia and reperfusion of rabbit hindlimb: implications for therapeutic intervention. AB - Peripheral ischemia was induced in the rabbit by occlusion of the left iliac artery for 6 hr, followed by 24 hr of reperfusion. Biochemical and morphological investigations were performed to evaluate the extent of vascular and tissue injury. Blood samples for plasma enzyme determinations (creatine kinase (CK) and lactate dehydrogenase (LDH) activities) were obtained at times t = 0, t = 6, t = 30 hr. Plasma CK and LDH activities in ischemic animals were approximately twice as high as those in sham-operated animals at the end of reperfusion, although no difference was observed at the end of the period of ischemia. Morphological and morphometric analysis of extensor digitorum longus muscle from ischemic animals showed a reduction in the number of patent capillary vessels per muscle fiber (1.54 +/- 0.1 and 1.04 +/- 0.09, P < 0.05, in sham and ischemic groups, respectively; mean +/- SEM). In addition, the number of microvilli on endothelial surfaces were considerably increased in the ischemic group (0.14 +/- 0.02 and 0.41 +/- 0.01 microns -2, P < 0.001, in sham and ischemic groups, respectively). A great number of adhered leucocytes were found on the vessel surface with some leucocytes having migrated through the vessel wall. Microcirculatory damage was accompanied by the formation of microthrombi which sometimes occluded the entire vessel lumen. The infusion of 1 mg/kg/hr of cloricromene for 6 hr prevented ischemic injury in microvessels and also prevented swelling of muscle mitochondria. In the treated group the number of patent capillaries per muscle fiber was very similar to that found in sham-operated animals (1.49 +/- 0.08; P < 0.01 vs. ischemic control). In conclusion, several different cell types are involved in the pathophysiological changes which occur in microvessels during ischemia/reperfusion injury. Pharmacological interventions, which inhibit the interactions of blood cells with endothelium, may be of value in the treatment of peripheral ischemia/reperfusion injury. PMID- 8412854 TI - Microvascular endothelial cell shape and size in situ. AB - To estimate total cleft length per unit surface area, previous studies assumed that endothelial cell clefts were oriented randomly with respect to the axis of the microvessel (Bundgaard and Frokjaer-Jensen, Microvasc. Res. 23, 1-30, 1982). In the present study, silver precipitation along the intercellular clefts of capillary endothelium ("silver lines") in the frog mesentery allowed observation of cleft orientation as well as estimation of cell shape, cell area (CA), and cleft length per unit area (CL). In all vessels, the endothelial cells were highly elongated polygons, usually hexagonal, which were oriented along the vessel axis. The clefts were highly oriented with a preferred orientation which was parallel to the vessel axis. Clefts exhibited very little local meandering, with a contour length only 7% greater than endpoint-to-endpoint length. Therefore, the assumption of random cleft orientation was not valid, and its use yielded an overestimate of CL. New estimates of CL were 0.16, 0.12, and 0.12 microns-1 for arterial, true, and venous capillaries, respectively. Cell lengths (mean +/- SD, n) were 135 microns (+/- 28, 79) in arterial capillaries, 98 microns (+/- 28, 19) in true capillaries, and 139 microns (+/- 20, 21) in venous capillaries. PMID- 8412855 TI - Human lung microvessel endothelial cells: isolation, culture, and characterization. AB - The pulmonary vasculature is of great physiological/pathological significance. We have isolated and cultured microvessel endothelial cells (HuLEC) from lung tissue obtained from lung transplant recipients by modification of published methods. Pure cultures of HuLEC were isolated by mechanical disaggregation of the tissue prior to sequential dispase and trypsin digestion to obtain microvessel fragments. Magnetic beads (Dynabeads) coated with Ulex europaeus agglutinin-1 were then used to enhance the purity of cultures at the first passage. HuLEC formed contact-inhibited "cobblestone" monolayers on gelatin and fibronectin substrates and capillary-like "tubes" on Matrigel and accumulated acetylated low density lipoprotein. Immunofluorescent characterization of these cells revealed the presence of von Willebrand Factor, angiotensin-converting enzyme, and thrombomodulin and the expression of antigens for the endothelial cell-specific monoclonal antibodies EN4, PAL-E, and H4-7/33. The endothelial origin of these cells was confirmed by the demonstration of the cell adhesion molecules, platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31), and E-selectin (endothelial leukocyte adhesion molecule-1/ELAM-1) upon stimulation with TNF alpha. These cells should provide a useful tool for studying various aspects of pathology and biology of the pulmonary microvasculature in vitro. PMID- 8412856 TI - Teenage pregnancies--the challenge of prevention. Sir William Power Memorial Lecture 1992. PMID- 8412857 TI - Textbooks of midwifery--what do they tell us about professional attitudes to pregnant women? PMID- 8412858 TI - "Curling and cuddling" abdominal exercises during and after pregnancy. PMID- 8412859 TI - Case study: bulimia in pregnancy. PMID- 8412860 TI - Midwifery care in The Netherlands. PMID- 8412861 TI - A new curriculum in The Netherlands. PMID- 8412862 TI - Dutch and UK breast-feeding practices compared. PMID- 8412863 TI - Words do matter. PMID- 8412864 TI - HIV and AIDS in childbirth: are midwives responding to the needs of women? The Mabel Liddiard Memorial Lecture, 1993. PMID- 8412865 TI - Open learning--identifying the demand within midwifery education. PMID- 8412866 TI - Post-registration education and practice--where are we now with PREP? PMID- 8412867 TI - Learning contracts: an innovative approach within a statutory midwifery refresher course. PMID- 8412868 TI - The quality question in midwifery education. PMID- 8412869 TI - Rise in caesarean rate in Britain. PMID- 8412870 TI - Medical specialization in Europe: the way forward. Promotion of educational quality. AB - The medical specialties are being intensively reconsidered in the countries of Europe. The important need is for promoting improvement of educational training programmes. Evaluation and improvement of specialist training programmes is the priority and not the setting up of qualifying examinations. The first necessity is to safeguard 20 years of evolution in postgraduate training. Support for the educational process is essential, to a much greater extent than occurs at present, if standards are to be improved and the confidence of the public is to be retained. The EC medical education system is the only existing international structure in medical education which is controlled by law, and is on that basis alone of the greatest interest. PMID- 8412871 TI - The teaching of medicine at the University Centre for Health Sciences Yaounde, Cameroon: its concordance with the Edinburgh Declaration on medical education. AB - The University Centre for Health Sciences (UCHS) or as it is referred to in French 'Centre Universitaire des Sciences de la Sante' (CUSS), became functional in 1969 with the enrollment of the first group of students. The objective of this training programme was to produce a scientifically sound, multipurpose doctor who would be fully operational in a rural setting with minimal equipment and supplies (Monekosso 1970, 1972). The graduate had to be able to adapt readily to new situations and improvise whenever possible, calling for a high degree of competence and initiative. The training strategies adopted by UCHS in 1969 which met this requirement were later found to be in close concordance with the tenets of the World Conference on Medical Education held in Edinburgh in 1988, the Edinburgh Declaration. While some of the terminology may not have been worked out at the time, the programme developed embraced some new concepts hitherto untried or undeveloped: the problem-solving approach in the first to the sixth year; an integrated teaching approach during the first to sixth year of medical training; an integrated medicine internship in district hospitals in the sixth year; a community-based training approach throughout the training; team training of three different health professionals; competency-based training; health services linked research; health services linked training (Monekosso & Quenum 1978). The concordance of this programme to the Edinburgh Declaration is of great interest in realizing the World Federation for Medical Education programme and implementing the Edinburgh Declaration. The involvement of the three innovative medical schools in the planning stage of the curriculum explains this concordance. PMID- 8412872 TI - Selection of Auckland medical students over 25 years: a time for change? AB - The selection procedures used in Auckland have been reviewed, and the characteristics of those admitted over 25 years analysed. Students are admitted either as school-leavers, mature entrants, or through an affirmative action scheme. A further small number are admitted as part of overseas development assistance. School-leavers are invited for interview on the basis of their academic achievement. Mature students and the affirmative group must have a minimum acceptable academic standard, with the interview playing a dominant role. Two thousand four hundred and forty-eight students have been admitted. The mean age was 18.6 years, and 39.7% were women. Over one half of the students had a parent who had attended university and 13% had a medical parent. One in ten students failed to complete the course, academic failure and withdrawal being of equal importance. The high loss seen in the affirmative group was due to academic failure and has led to the introduction of extra tuition and support for these students. The emphasis on academic achievement by school-leavers has excluded many applicants with outstanding personal qualities. The academic staff has therefore decided to modify the selection procedure, the final rank order of these applicants being based on their personal attributes and life experiences. PMID- 8412873 TI - Medical school entrants: semi-structured interview ratings, prior scholastic achievement and personality profiles. AB - The Faculty of Medicine at Monash University has decided to take personal qualities as appraised by semi-structured interviews into account alongside academic merit for selection of undergraduate students. To develop competence in these techniques the Faculty interviewed entrants, rather than applicants, in 1991 and 1992. Interviewing panels consist of three members--a member of the Faculty of Medicine, a member of the association of Monash Medical Graduates Inc. and an outside person who is not involved in medicine. The qualities appraised fall into four fields--quality of motivation, appropriateness of cognitive style, appropriateness of interpersonal style and communication skill. Interviewers are also asked to rate the overall suitability of the interviewee for medical studies and medical practice. A pilot study has also been run to explore the value of a psychometric test, the California Psychological Inventory (CPI), as another adjunct to the selection procedure. In this report interview ratings are analysed in relation to both prior scholastic results and scores obtained on the CPI. The study showed that the interview scores have little overlap with prior scholastic results. The associations between interview ratings and CPI scores were often highly significant from a statistical point of view and thus confirm through a different instrument that the interviews do meaningfully appraise personal attributes. However, the correlations are modest in terms of the amount of variance which is shared, suggesting that the CPI could be advantageously used as another component of our selection process.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412874 TI - Surgeons' personalities: the influence of medical school. AB - Does medical school shape surgeons' personalities? This research retrospectively compares personality test results of 17 surgical and 44 non-surgical specialists trained at the same medical school. Fifty-five standardized personality measures at two developmental periods (the outset and the conclusion of medical school) revealed no pronounced differences between surgeons and other specialists. Interviews with 30 academic doctors (10 surgeons, 10 anaesthesiologists and 10 psychiatrists) explore the apparent discrepancy between personality tests scores and commonly held opinions that surgeons' personalities differ from other doctors. PMID- 8412875 TI - Behavioural assessment of the effectiveness of a communication programme for medical students. AB - This study evaluated the effectiveness of a communication programme taught to medical students at the University of Melbourne in their preclinical years. The effectiveness of the programme was assessed by comparing videotaped history taking interviews completed by a cohort of first-year clinical students in 1986, who had not undertaken the communication programme, with a similar cohort of first-year clinical students in 1992 who had undertaken the programme. The students from the 1986 cohort who had not undertaken communication training in their preclinical course completed their videotaped interviews as part of the experimental evaluation of a consulting skills training programme carried out in 1986-87. A comparison of ratings given by two experimentally naive, independent observers revealed that the 1992 student cohort demonstrated some significantly better skills at questioning and facilitating communication with patients. By contrast, the 1986 student cohort showed significantly greater skills at maintaining relevance in their interviews and greater capacity to explore patients' psychosocial concerns. These data suggest that students acquire the most effective interview skills when interacting with patients during their clinical training. PMID- 8412876 TI - Teaching clinical methods to medical students. AB - This paper demonstrates that it is feasible to teach clinical methods in general practice and describes the organization of an appropriate attachment. Willing practitioners, properly briefed, are competent to undertake clinical methods teaching and the attachment provides satisfaction to both students and teachers. It is possible to provide in primary care the elements which seem to be the key to this outcome--protected time, teaching in very small groups, direct observation and concentration on systematic examination rather than on particular physical signs. There is a strong case for the promoting of clinical methods teaching by general practitioners. PMID- 8412877 TI - Medical students' experiences with and perceptions of chronic illness prior to medical school. AB - Concerns have been expressed about the failure of the medical curriculum to address the health-care needs of the chronically ill. It has been shown in the literature that medical students develop cynicism and negative feelings towards chronic illness as they progress through their training, perhaps as a result of the attitudes and frustrations of their teachers. What has been inadequately addressed are the experiences with and perceptions about chronic illness that medical students have before entering medical school. Some recommendations are made for curriculum changes based on the findings reported. PMID- 8412878 TI - International health teaching: a survey of 100 medical schools in developed countries. AB - The survey reported here was designed to assess the availability of international health teaching to medical students in developed countries. For the purpose of this enquiry international health teaching was defined as any instruction on comparative morbidity or mortality, service provision, demographic change and disease prevalence in non-industrialized developing countries. The results indicated that this topic is not regarded as being of high priority in the majority of institutions. PMID- 8412879 TI - Medical student abuse: perceptions and experience. AB - A questionnaire containing 18 vignettes of common clinical educational situations with potentially abusive treatment of medical students and a 10-item attitude assessment about abusive behaviour were administered to the first- and fourth year medical students at a mid-west US university medical school. The first- and fourth-year groups did not differ significantly on perceived abusiveness of most of the vignettes, although several of the individual vignettes were perceived significantly differently by the two groups. As hypothesized, the fourth-year students had experienced such situations more frequently. Attitudes towards abusive behaviour did not differ between the two groups. The authors contrast teaching interactions perceived as educationally useful and not abusive with those seen as abusive and not useful and offer explanations for the differences observed. Finally, the possible implications of the results for medical education are discussed. PMID- 8412880 TI - A solution to the cueing effects of multiple choice questions: the Un-Q format. AB - Although the cueing effects inherent in conventional multiple choice questions (MCQs) present serious limitations, this format continues to dominate testing programmes. The present study was undertaken to estimate the effects of cueing when MCQs are used to test medical students, and to evaluate the reliability, validity and feasibility of an alternative testing format. Equivalent items in both MCQ and open-ended, or uncued (Un-Q), formats were administered to 34 third- and fourth-year medical students. The students' mean % correct score on the MCQs was 11 percentage points higher than their mean level of performance on equivalent Un-Qs. When a second set of more difficult items was administered to 16 of these students, their mean performance on the MCQ items was 22 percentage points higher than their performance on equivalent Un-Qs. The results support the feasibility of large group administration of tests constructed in an open-ended format that can be scored by computer. Not only is this format equally reliable and economical when compared with the MCQ, but it also provides important advantages that strengthen its face validity. The Un-Q format can be used to test either simple recall or certain higher level problem-solving skills that cannot be tested by MCQs. Even more important, the results also suggest that the Un-Q format may be a more effective discriminator of academically marginal examinees. PMID- 8412881 TI - Standardized patients: a long-station clinical examination format. AB - Data from the first 20 periods of a long-station clinical performance examination for a 4-week required clerkship in family medicine were examined in order to assess the reliability and validity of the examination. Data from 304 students were examined for station, case scenario and examiner effects and results compared to short-station formats. A significant examiner effect was found but there were no differences in student performance for station or case scenario. These findings reflect examiner specificity cited in the literature for short station examinations, but not case specificity. The source of variability for this examination appears to be primarily examiner effect. There was a significant correlation between student scores on the two cases, and raters tended to rank order students similarly in spite of variability in mean rater score. Scores on the CPE correlated with other measures of clinical performance as well as other methods of student evaluation for the clerkship providing some evidence for construct and criterion-related validity. CPE cases were developed from clerkship objectives but examination of the test blueprint revealed some gaps in the extent to which the CPE covers the course content. CPE developers are working to increase interrater reliability through examiner training and further standardize case scenarios through check-lists and patient training. Additional cases are being developed to increase the content validity of the examination. PMID- 8412882 TI - Dissatisfied patients: improving general practitioners' initial reactions. AB - General practitioners often have difficulty in dealing with dissatisfied patients. One underlying reason could be the disturbed relationship between the doctor and the dissatisfied patient. A training course has been developed taking the relationship as a starting-point. Based on Watzlawick et al.'s theory on communication GPs have been trained to react to a dissatisfied patient on a relational level ('Are you dissatisfied with my treatment?') rather than on a contents level ('How long have you been suffering from this?'). This method seeks to improve the relationship and the satisfaction of both doctor and patient. Three types of initial reaction to dissatisfied patients were offered to four groups of GPs (19 trainees in general practice and 19 trainers in general practice). Pre- and post-measurement were executed by means of registering the initial reactions on videorecorded vignettes of re-enacted dissatisfied patients. Subsequently the reactions were categorized blind by two judges. The 12 possible categories can be subdivided into categories primarily aimed at the contents or primarily aimed at the relationship. The results show that, as compared to the pre-measurements, GPs more frequently use empathic reactions and reactions in which they bring their own actions up for discussion. The number of responses in which doctors ask a further clinical question or in which GPs expect a solution whether from themselves or from others, decrease. It is concluded that the course appears to change for the better the GPs' initial reaction to dissatisfied patients. PMID- 8412883 TI - Allocating pre-registration house officer posts. PMID- 8412884 TI - Obstetrics in crisis? PMID- 8412885 TI - Whither genetic services? PMID- 8412886 TI - Steroid hormone receptors and breast cancer. PMID- 8412887 TI - Oral contraceptives--can we make them more effective? PMID- 8412888 TI - Oestrogen and progesterone receptor assays in breast tumours. The Prince Henry's Hospital experience, 1983-1990. AB - OBJECTIVE: To present and analyse the results of eight years of experience (1983 1990) in breast tumour receptor analysis. DESIGN: All female primary breast tumour samples received (4683) were analysed for seasonal variation, patient age, relative risk index, oestrogen receptor (ER) and progesterone receptor (PR) status, ER and PR status as a function of age, ER and PR levels as a function of age, and ER and PR levels as a function of month of analysis. SETTING: The assays were done at the Medical Research Centre, Prince Henry's Hospital, Melbourne, as a non-profit service to surgeons, oncologists and pathologists. RESULTS AND CONCLUSIONS: The numbers of samples referred for assay increased progressively each year, from 473 in 1983 to 1097 in 1990, but the receptor status (ER +/-, PR +/-) appeared not to vary from year to year. ER+PR+ tumours were the most common in all age groups, steadily increasing from between 50% and 60% in premenopausal women to 70% or more in those aged over 80. In postmenopausal women, levels of ER in ER+ tumours were three times those in premenopausal women; PR levels in PR+ tumours, however, were bimodal, with higher levels in the age groups 35-49 and 70 89 years than in women aged 50-69 years. No significant seasonal variation was seen, and the overall patterns of receptor status are similar to those seen in Northern hemisphere studies. PMID- 8412889 TI - Menopausal symptoms in Australian women. AB - OBJECTIVES: To describe Australian-born women's experience of symptoms during the natural menopause transition and the relative contribution of menopausal and health status, social factors and lifestyle behaviours. DESIGN: A community based cross-sectional survey by telephone interview was carried out on a randomly derived sample of Melbourne women. PARTICIPANTS: The participants were 2000 Australian-born women, aged between 45 and 55 years. OUTCOME MEASURES: A list of 22 symptoms was used. Explanatory variables were: sociodemographic variables; menopausal and health status; lifestyle behaviours; attitudes to ageing and to menopause. RESULTS: A 70% response rate was achieved for eligible women who could be contacted during the study. Premenopausal women were the least symptomatic and perimenopausal women the most symptomatic. Factor analysis found seven common factors from the 22 symptoms studied. Menopausal status based on menstrual history was significantly related to two groups of symptoms: vasomotor symptoms, which increased through the menopausal transition; and general somatic symptoms which were more frequent in the perimenopause. Analysis of variance of factor scores found fewer symptoms with increasing years of education, better self-rated health, the use of fewer non-prescription medications, the absence of chronic health conditions, a low level of interpersonal stress, the absence of premenstrual complaints, not currently smoking, exercise at least once a week, and positive attitudes to ageing and menopause. CONCLUSIONS: Many factors unrelated to hormonal changes contributed to the symptoms. Longitudinal investigation is needed to determine the relative importance of hormonal, psychosocial and lifestyle variables in the aetiology of mid-life symptoms. PMID- 8412890 TI - Diagnostic procedures and health outcomes. Upper gastrointestinal tract investigations in the elderly. AB - OBJECTIVES: To examine the use of upper gastrointestinal endoscopy in people 65 years of age and over in New South Wales, and compare its use between geographic areas, sex and age groups. Also, to investigate the relationship between endoscopy use and peptic ulcer mortality. DESIGN: A retrospective review of upper gastrointestinal tract (GIT) endoscopy hospital separation data for NSW from 1986 to 1989-1990 and peptic ulcer mortality data from 1979 to 1989. MAIN OUTCOME MEASURES: Rates per 1000 population aged 65 years and over were calculated for upper GIT endoscopy for each of the four-year age groups and by sex. Peptic ulcer and other digestive system disease rates for endoscopy patients were determined. Correlations between hospital standardised separation rates for endoscopy and standardised mortality rates for gastric cancer and peptic ulcers were calculated. Age and sex standardised death rates were calculated for each year between 1979 and 1989, and changes in death rates over time were determined by regression analysis. RESULTS: The rate of endoscopies increased significantly (P < 0.001) over the period 1986 to 1989-1990, with the highest rate and greatest increase occurring in patients aged 70 years and over. Men had significantly higher endoscopy rates than women (P < 0.001). Geographic variations in the use of endoscopy were found and were not explained by geographic differences in the proportion of peptic ulcers diagnosed, deaths from peptic ulcers or malignant neoplasms. Despite the increase in endoscopies performed, there was no corresponding significant increase in the rate of peptic ulcers detected. Between 1979 and 1989 peptic ulcer mortality rates for women over 74 years of age increased significantly (P < 0.05); the rate for men did not change. CONCLUSION: Many endoscopies are performed in people aged 65 years and over in NSW and there has been a substantial increase in endoscopy rates in recent years. While the increase in upper GIT endoscopy may have resulted in a number of health benefits, it does not appear to have resulted in a reduction in peptic ulcer related mortality. PMID- 8412891 TI - Community Health and Medical Practitioners Scheme: providers evaluate a pilot program of integration of services. AB - OBJECTIVE: To set up and evaluate a pilot scheme integrating salaried community health centre staff and fee-for-service medical practitioner services (CHAMPS). DESIGN: Preliminary interviews with both groups established the aims, logistics and financial arrangements of the project. The community health centre provided staff and the general practitioners provided premises and administrative services. Follow-up interviews evaluated the scheme and made recommendations. SETTING: A New South Wales country town, population 24,000, with 25 general practitioners and 23 community health centre professionals. RESULTS: Six general practitioners and 23 community health professionals determined the aims to be: improved access for patients to community health services; improved liaison between the two groups of providers; and a broadening of services offered at general practice locations. Two dietitians and three mental health workers were rostered for half a day per week in four general practices for six months. The dietitians continued after the project finished, but the mental health workers did not. The five community health staff, five of the general practitioners originally interviewed and six other general practitioner participants cited the major benefits as increased communication between providers and improved access for patients, and the major difficulties as lack of appropriate equipment and organisational logistics. CONCLUSIONS: The providers believe that the project succeeded in improving access to community health services and liaison between professionals. For future projects they recommended better communication, firmer role delineation and better planning for space and equipment. PMID- 8412892 TI - Parkinson's disease. AB - The prevalence of Parkinson's disease rises with age. The clinical features can be the typical ones or those associated with the "late-onset" type, with more gait and balance problems and a mild progressive dementia. Other disorders, such as stroke and essential tremor, may be mistaken for Parkinson's disease, or multiple conditions may be present which modify the response to treatment. The treatment of choice is a low dose of one of the L-dopa-decarboxylase inhibitor drugs. Other agents have a high incidence of side effects, particularly confusion, which also occurs with L-dopa in higher doses. PMID- 8412893 TI - Getting "value for money". Measuring the quality and outcome of general practice care. PMID- 8412894 TI - Influenza vaccination in Victoria, 1992. AB - OBJECTIVES: To determine the rates and patterns of vaccination against influenza in the Victorian population. Further, to determine rates of vaccination against influenza in residents of Victorian nursing homes and hostels. DESIGN: Between May and June of 1992 we conducted a telephone survey of 537 systematically selected Victorian residents. We obtained information on age, sex, vaccination status, and level of risk for the complications of influenza. We conducted a separate postal survey of 143 randomly selected Victorian nursing homes and hostels to establish rates of vaccination within these institutions. PARTICIPANTS: Residents of Victoria listed in the White Pages telephone directories in both metropolitan and rural areas. We made calls during working hours to both residential and business numbers, and spoke to the first person who answered the telephone. Participants in the nursing home survey were the matrons or directors of nursing of randomly chosen facilities in the State, who filled out questionnaires relating to their residents. RESULTS: The vaccination rate against influenza in the whole population was 13.3%. This and all other figures quoted are adjusted for age, unless otherwise specified. There was a significant difference in vaccination rates between rural (9%) and metropolitan (16%) areas (P < 0.05). Using the National Health and Medical Research Council (NHMRC) guidelines, 29% of the population had at least one indication for vaccination. Of people at low risk for complications of influenza, 7.8% were vaccinated. Of people with one or more specific indications for vaccination, 29.4% were vaccinated. Vaccination rates increased with increasing number of indications and with increasing age, approaching 45% in people over 65 years and 50% in people with three or more indications for vaccination. The postal survey of nursing homes and hostels revealed that a mean of 52% of residents per facility were vaccinated against influenza. This figure varied between 45% in rural facilities and 53% in metropolitan facilities. CONCLUSIONS: We estimate that 550,000 doses of influenza vaccine were used in Victoria in 1992. Approximately 50% of these were given to people at low risk for complications of influenza. These findings highlight the need to target high risk groups for vaccination, reassure the general public, and educate health care professionals. PMID- 8412895 TI - Who will deliver the next generation? PMID- 8412896 TI - Delivery of traditional hospital services to patients at home. AB - The high cost of traditional inpatient care, advances in the delivery of depot antibiotics and narcotics, and the demands of patients for realistic alternatives to frequent hospitalisation have led to programs for the provision of acute hospital care in the home. We review the progress of overseas hospital-in-the home programs and the issues that such service delivery has raised for clinicians and administrators. The potential application in Australia is examined and priorities for preparative research are suggested. It seems inevitable that for a select number of conditions in a select group of patients, home hospital services will become a reality in this country. PMID- 8412897 TI - Non-communicable diseases in the tropics. AB - The way of life in many developing countries in the tropics is in a state of transition--from traditional to "modern", with its associated industrialisation, urbanisation and cultural readjustment. Infectious diseases are giving way to non communicable diseases, such as cardiovascular diseases, adult onset diabetes, cancers, trauma and non-infectious respiratory diseases, as major causes of morbidity and premature mortality. The epidemiology, causality, prevention and treatment of these conditions is summarised in the context of developing countries. PMID- 8412898 TI - Familial hypobetalipoproteinaemia: a rare presentation to the lipid clinic. AB - OBJECTIVE: To report a case of familial hypobetalipoproteinaemia in a woman who presented after the incidental finding of marked hypocholesterolaemia during laboratory tests. CLINICAL FEATURES: An asymptomatic 37-year-old Lebanese woman presented to the lipid clinic with a serum total cholesterol concentration of 1.1 mmol/L, high density lipoprotein (HDL) cholesterol of 1.0 mmol/L, and triglycerides of 0.28 mmol/L. No secondary cause for the hypocholesterolaemia was established. INVESTIGATION AND OUTCOME: Her serum apolipoprotein B (apo B) levels were markedly reduced at 0.07 g/L. Except for one daughter (IV-4), all other family members including her husband (her first cousin) had apo B levels about 25% of normal. Daughter IV-4 had undetectable apo B levels. Family studies confirmed an autosomal dominant pattern of inheritance consistent with familial hypobetalipoproteinaemia. CONCLUSION: Familial hypobetalipoproteinaemia is a rare condition that should be considered in the differential diagnosis of hypocholesterolaemia. Absence of clinical features, autosomal dominant pattern of inheritance, and reduced apo B levels suggest the diagnosis. PMID- 8412899 TI - Gene therapy and related novel forms of treatment. PMID- 8412900 TI - Ethics, distribution and Oregon. PMID- 8412901 TI - Medical journals and the popular media. The International Committee of Medical Journal Editors. PMID- 8412902 TI - Medical litigation and the cost of premiums for obstetricians. PMID- 8412903 TI - Sexual transmission--a major HIV risk factor in people using drugs and alcohol. PMID- 8412904 TI - Rheumatic fever. PMID- 8412905 TI - Medicine in tropical Australia: the quality and price of snake antivenoms. PMID- 8412906 TI - Hepatitis C virus in the central province of Papua New Guinea. PMID- 8412907 TI - What south Australian GPs think about smart cards. PMID- 8412908 TI - The plagues of Egypt: what killed the animals and the firstborn? PMID- 8412909 TI - Liability for failure to diagnose HIV infection and AIDS. PMID- 8412910 TI - Cigarette smoking and teenagers: one novel approach. PMID- 8412911 TI - The tobacco company Rothmans Holdings. PMID- 8412912 TI - Telemedicine. A health care system to help Australians. PMID- 8412913 TI - The health care of young adults with cerebral palsy. AB - OBJECTIVE: To evaluate the health status and provision of health services to young adults with cerebral palsy. RESULTS: Ninety-seven per cent had ongoing health problems. Consultations with most health professionals declined markedly after leaving school. CONCLUSIONS: Young adults with cerebral palsy have considerable, continuing impairment and disability. Lack of contact with health services after they leave school may adversely affect their health status. PMID- 8412914 TI - Laparoscopic hysterectomy: a series of 100 cases. AB - OBJECTIVE: To introduce a new method of hysterectomy using the laparoscope and endoscopic stapling equipment. DESIGN: A prospective study of a series of patients. SETTING: Specialist private practice at two private hospitals. PATIENTS: Patients were accepted for laparoscopic hysterectomy except when clinical or ultrasound examination indicated that conventional vaginal or abdominal hysterectomy would be more appropriate or less hazardous. RESULTS: The procedure took longer than conventional surgery but was followed by an accelerated recovery, both in terms of inpatient stay and a short convalescence. CONCLUSIONS: Laparoscopic hysterectomy is an alternative to conventional hysterectomy with apparent advantages. Patient recovery is generally faster. With increasing experience, complications of the procedure are reduced to levels that are comparable with those of conventional surgical hysterectomy. PMID- 8412915 TI - The contribution of the undergraduate rural attachment to the learning of basic practical and emergency procedural skills. AB - OBJECTIVE: To assess the effect of a four-week rural attachment on the knowledge and competency of medical students in basic practical and emergency procedures. DESIGN: A before and after comparison of self-reported level of competence in 72 basic, emergency, diagnostic and therapeutic procedural skills. PARTICIPANTS: Eighty-five final year medical students at the University of Western Australia in 1991. OUTCOME MEASURE: A student was considered competent if he or she claimed to be able to do a procedure either alone or with assistance. RESULTS: Over 50% of students were competent in 57 procedures after the attachment compared with 37 procedures before it. There were 26 procedures in which more than 20% of students increased their competence. CONCLUSION: A higher priority needs to be paid to the undergraduate teaching of procedural skills. Rural attachments can play an important part in training medical students in the practical skills required of interns. PMID- 8412916 TI - The experience of a single Australian paediatric oncology unit. 1000 patients 1964-1987. AB - OBJECTIVE: To determine the survival for children with malignant disease diagnosed in the period 1964-1987 and treated in a single paediatric oncology unit. DESIGN: Records of patients treated by the Department of Haematology and Oncology at the Prince of Wales Children's Hospital were reviewed to determine the survival of children with cancer according to decade of diagnosis and diagnostic group. PATIENTS: Patients were eligible for the study if referred for treatment at or soon after diagnosis of malignancy. One thousand patients were treated during the study period. There were 363 with acute lymphoblastic leukaemia (ALL), 126 with tumours of the central nervous system (CNS), 86 with acute non-lymphoblastic leukaemia (ANLL), 81 with lymphoma, 79 with neural crest tumours, 69 with renal tumours, 66 with bone sarcomas, 53 with soft tissue sarcomas, and 77 with various other diagnoses. Age range was one day to 20.75 years. INTERVENTIONS: Treatment included surgery, radiotherapy and chemotherapy in a variety of protocols. RESULTS: Ten-year survival for the 1960s, 1970s and 1980s was 15%, 51% and 64% respectively (P < 0.001), excluding tumours of the CNS. From 1985 onwards, actual survival at five years has been 79%. Survival from Wilms' tumour and Hodgkin's disease remained high throughout the study period, and significant improvement in survival occurred with ALL, non-Hodgkin's lymphoma (NHL) and osteogenic sarcoma. Survival remained poor with neuroblastoma and ANLL. CONCLUSIONS: Significant improvement in outcomes for childhood malignancy has been achieved over the last three decades, with five-year survival currently at 79% (excluding tumours of the CNS). Some diagnostic groups have had only small improvements in outcome and require new strategies. PMID- 8412917 TI - Postpartum haemorrhoids--evaluation of a cooling device (Anurex) for relief of symptoms. AB - Postpartum haemorrhoids are a common problem, for which the recently marketed cooling device, Anurex (Roussel Uclaf Australia, Pennant Hills, NSW), was evaluated in 129 women randomly allocated to treatment and control groups. There was no difference in the rate of resolution of pain between the groups. The test group estimated a greater duration of pain relief, but this was not statistically significant. Anal bleeding and itching were not significantly reduced in the test group. There was no difference between discomfort scores related to application of the device in the groups, but a significant number in the test group encountered difficulty in insertion of the device. It is concluded that the introduction of the Anurex device as a standard method of treatment of postpartum haemorrhoids is not justified. PMID- 8412918 TI - For the new millennium: control of helminth diseases throughout the world. AB - In the latter half of this century there has been a dramatic change in the treatment of helminthic infections. Two factors have contributed to this: an understanding of the major differences between helminths and other infectious agents, and the development of effective low-cost, single-dose, broad spectrum drugs. A program of administration of these drugs to school-aged children in developing countries is now being implemented and will result in a significant improvement in their health at a crucial time in their growth and development. PMID- 8412919 TI - Worms in Australia. AB - Fortunately, Australia does not have many of the parasitic worm infections, such as filariasis, onchocerciasis and dracunculiasis, that are important on a global scale. Nevertheless, several medically important parasitic worms are endemic in Australia. Intestinal nematodes occur mostly in the tropical north although threadworm is ubiquitous. Hydatid disease and tapeworm infections occur especially in sheep and cattle farming regions. PMID- 8412920 TI - Sudden unexpected cardiac death among Tasmanian men. AB - OBJECTIVE: To examine sudden unexpected cardiac death (SUCD) in Tasmanian men and to assess the contribution of these deaths to differences in rates of mortality from ischaemic heart disease (IHD) between regions within Tasmania. DESIGN AND SETTING: Descriptive epidemiological study based in the community. SUBJECTS: Male residents of Tasmania aged 30 to 69 years who died from IHD suddenly and unexpectedly between 1987 and 1989. MAIN OUTCOME MEASURES: Rates and proportions of sudden cardiac death in men who had no prior overt signs of IHD, validated by necropsy and information from the attending doctors. RESULTS: SUCD accounted for approximately 24% of deaths from IHD among men aged 30 to 69 years in Tasmania. The ratios of observed to expected numbers of deaths occurring among manual and nonmanual workers were similar whereas there was an excess of events among men aged less than 65 years who were not working. Examination of data from necropsy reports revealed that 32% of cases showed evidence of previous "silent" myocardial infarction and 63% showed severe coronary artery disease in two or more vessels, with a further 28% having severe single vessel disease. The contribution of SUCD to total mortality from IHD varied from 20% in the north west region to 25% in the south and 26% in the north, but the limited number of events makes it uncertain whether the variation in the rate of SUCD is significantly different from that for total mortality from IHD. CONCLUSIONS: The proportion of deaths from IHD among men in Tasmania which are sudden and unexpected and the associated necropsy findings are consistent with those described in other population based studies of sudden cardiac death. Non participation in the workforce was a risk factor in SUCD. As yet, we cannot distinguish whether the higher mortality rates from IHD among men in northern regions of Tasmania (P < 0.01) were due to differences in SUCD or whether the same rate of SUCD obtains in all regions of the State and the differences in mortality from IHD reflect variation in non-sudden deaths and deaths in people with overt IHD. PMID- 8412921 TI - Data collection and changing health care systems. 1. United Kingdom. AB - The adoption of "Read Clinical Codes" for computerised patient records could profoundly alter the nature and quality of Western medicine in the next decade. The increasing awareness of the need for a standardised coding system has led to the funding of a pilot project to test Read codes in Australian general practice. Read codes are a comprehensive nomenclature of clinical terms incorporating over 100,000 codes in a structured hierarchical form. Designed by a general practitioner, they are now owned, controlled, and developed by the British National Health Service (NHS). Selected as the basis for clinical coding across the NHS, they form the cornerstone of computerised patient records. Computer use has been encouraged in general practice in the UK, with financing schemes and functional inducements resulting in 70% of practices being at least partly computerised, and 84% of these using Read codes. Their promotion has been backed by a major development program to broaden the codes to include all clinical specialities, nursing, and professions allied to medicine. The codes will require significant adaptation for Australian use, including the development of an administrative chapter and a pharmaceutical classification. The impact of information management systems on health care in the UK has relevance for the continuing development of the Australian National Health Information Strategy and for future record keeping in general practice in Australia. If the trial proves successful, the adoption of Read codes as a standard for information management in patient medical records will need to be considered. PMID- 8412922 TI - Data collection and changing health care systems. 2. New Zealand. AB - Radical changes planned in the New Zealand health system aim to improve its cost effectiveness, quality and consumer responsiveness. These changes will take place despite a paucity of data on the use of resources and outcomes. Data collection systems are to be introduced into hospitals and primary care with the use of the Read clinical codes (RCC) in addition to ICD-9-CM (International classification of disease--clinical modification) and ICPC (International classification of primary care). This paper discusses the proposed changes to the New Zealand health care system and describes their effect on the present and future data collection methods in general practice. PMID- 8412923 TI - The Somogyi effect. Has it ever existed and what harm has it caused? PMID- 8412924 TI - The Somogyi effect. The paediatric view. PMID- 8412925 TI - The problem of hypoglycaemia--Somogyi or not. PMID- 8412926 TI - Ketorolac and renal failure. Adverse Drug Reactions Advisory Committee. PMID- 8412927 TI - Ketorolac. PMID- 8412928 TI - Metronidazole resistant Trichomonas vaginalis in Brisbane. PMID- 8412929 TI - Good grief. PMID- 8412930 TI - Lichtenstein herniorrhaphy. PMID- 8412931 TI - Qualitative research in medical practice. PMID- 8412932 TI - Oral sodium phosphate bowel preparation for colonoscopy. PMID- 8412933 TI - Fluoxetine (Prozac) PMID- 8412934 TI - The doctor as expert witness. PMID- 8412935 TI - Evidence for meningitis in Ross River virus infection. PMID- 8412936 TI - Ozone therapy in AIDS--truly innocuous? PMID- 8412937 TI - A case of acute hepatitis E in Victoria. PMID- 8412938 TI - Epidemiology of Haemophilus influenzae type b (Hib) meningitis in South Australia. PMID- 8412939 TI - Society is becoming increasingly violent. PMID- 8412940 TI - A preventive health questionnaire in one general practice. PMID- 8412941 TI - A letter from Patrick White. PMID- 8412942 TI - Why doctors and nurses must not kill patients. PMID- 8412943 TI - Vitamin K and childhood cancer. PMID- 8412944 TI - Did Vietnam veterans get cancer from dapsone? PMID- 8412945 TI - A clinical school for north Queensland. PMID- 8412946 TI - The National Cancer Prevention Policy: will it sell itself? PMID- 8412947 TI - Achieving better health in Australia in the next five years. PMID- 8412948 TI - Smoking and the incidence of coronary heart disease in an Australian population. AB - OBJECTIVE: To estimate the impact of smoking on the incidence of coronary heart disease in Australia. Data collected for the WHO MONICA Project were used. DESIGN: Combined data from a community-based register of all suspected coronary events and a survey of risk factor prevalence in a random sample of the same population. SETTING AND PARTICIPANTS: All residents of the Hunter Region of New South Wales aged 35-69 years who had a first acute myocardial infarction or fatal heart attack (without a history of coronary heart disease) between 1 January 1986 and 31 December 1990. MAIN OUTCOME MEASURES: Acute myocardial infarction or coronary death, as defined by the WHO MONICA Project. RESULTS: Men who are current smokers are 2.9 times (95% CI, 2.7-3.1) more likely than non-smokers to have a first myocardial infarction or fatal heart attack, and for women the equivalent figure is 3.5 times (95% CI, 3.2-3.8), after adjusting for age. Current male smokers with a history of hypertension are 4.5 times more likely to have a coronary event (7.9 times in women) than are non-smokers without a history of hypertension. The age-adjusted excess rate was 566 per 100,000 per year in men and 373 per 100,000 per year in women. Smoking is a stronger predictor of coronary heart disease incidence than a history of hypertension (relative risk [RR] = 1.6 for men and 1.9 for women) or a known history of hypercholesterolaemia (RR not significantly different from 1). CONCLUSIONS: Cigarette smoking plays a more important role in the causation of a first myocardial infarction or fatal heart attack and appears to have more influence on the incidence of coronary heart disease in Australia than hypertension. PMID- 8412949 TI - Cell-mediated immunity in combat veterans with post-traumatic stress disorder. AB - OBJECTIVE: To explore immune function in patients with chronic post-traumatic stress disorder (PTSD). DESIGN: A case-control study using the standardised Cell mediated Immunity (CMI) Multitest. PARTICIPANTS: Cases were 25 Vietnam combat veterans undergoing treatment for clinically diagnosed PTSD. Controls had no diagnosed psychiatric disorders: 28 were civilians and 20 were servicemen who had had South-east Asian postings but no combat experience. MAIN OUTCOME MEASURES: The sum score, the number of reactions and the compound scores from the CMI Multitest were assessed as a measure of immune function. RESULTS: Psychometric tests distinguished between cases and controls at a highly significant level (P < 0.001). Veterans with PTSD had enhanced cell-mediated immunity compared with civilians (P = 0.008) and servicemen (P = 0.02). CONCLUSION: PTSD in combat veterans is associated with enhanced cell-mediated immune responsiveness. This could have wide implications for psychiatry and general medicine. PMID- 8412950 TI - Smoking in child family day care homes: policies and practice in New South Wales. AB - OBJECTIVES: To provide estimates of the numbers of New South Wales children in Family Day Care who may be exposed to environmental tobacco smoke while attending day care; to describe existing smoking policies; and to analyse these policies with the aim of providing guidelines for smoking policy in Family Day Care. SETTING: All 109 Family Day Care schemes in NSW. METHOD: Scheme coordinators were sent a questionnaire regarding the proportion of carers who smoked while caring for children; the nature, enforcement and experience of smoking policies; and barriers to implementation of a no-smoking policy. RESULTS: A mean of 10% of Family Day Care caregivers were reported to smoke while caring for children (range, 0-60%). An estimated 2045 children were potentially exposed to environmental tobacco smoke in the 86 schemes which provided this information. Thirty-five per cent of schemes had formal no-smoking policies. A range of advantages, disadvantages and perceived practical and legal barriers to implementation of a no-smoking policy in Family Day Care were described. Forty four per cent of schemes with no-smoking policies reported no implementation problems. CONCLUSIONS: There is considerable potential for exposure of children to environmental tobacco smoke in Family Day Care homes. There is legal support for Family Day Care caregivers not to expose children under their care to environmental tobacco smoke. A formal (and enforced) no-smoking policy should exist in every Family Day Care scheme, and a "top-down" directive is most likely to be successful. The issue of other smokers in the caregiver's household needs to be specifically addressed in any such directive. PMID- 8412951 TI - The adequacy of management of women with CIN 2 and CIN 3 Pap smear abnormalities. AB - OBJECTIVE: To assess the adequacy of management of Sydney women with Papanicolaou (Pap) smears showing cervical intraepithelial neoplasia (CIN) grades two and three. DESIGN: A prospective descriptive study of patient management. METHOD: All 206 general practitioners (GPs) who sent Pap smears which were reported as CIN 2 or CIN 3 to three Sydney laboratories between January and June 1990 were included in the sample. The GPs were contacted and management details for the women collected. The adequacy of management was evaluated by comparing it with management guidelines constructed for the study. Management steps were: notification of results, referral for colposcopy, biopsy, treatment and follow up. The number of women reaching each step, expressed as a proportion of those women who should have reached that step according to the management guidelines, was calculated. The points where management was inadequate were established and the reasons explored. RESULTS: All GPs provided management information about their patients. Ninety-nine per cent of women (95% confidence intervals [CI], 98% 100%) were informed of their Pap smear result and 94% (CI, 90%-97%) of the total sample subsequently underwent colposcopy. Ninety-three per cent (CI, 88%-97%) of the 160 women definitely requiring histological diagnosis had this and 98% (CI, 94%-100%) of the 140 women definitely needing treatment were treated. Following treatment (or equivalent if not indicated) 80% of women (CI, 74%-86%) were known to have had a follow-up Pap smear but only 31% (CI, 24-38%) of women needing follow-up colposcopy were known to have had it. At the time of the study (on average, 17 months after the index Pap smear) only 60% (CI, 54%-67%) of women were still being followed up. CONCLUSIONS: Virtually all women with the more severe cytological abnormalities are being referred by their GPs for further investigation and treatment. However, follow-up after treatment is often inadequate. The responsibility for this follow-up needs to be clarified, as well as communication about it between women, their GPs and gynaecologists. A cytology registry could increase the proportion of Sydney women known to be receiving follow-up. PMID- 8412952 TI - Undiagnosed post-traumatic stress disorder following motor vehicle accidents. AB - OBJECTIVES: To determine the pattern of emergence of post-traumatic stress disorder (PTSD) among motor vehicle accident victims and to examine the influence of PTSD on subsequent levels of disability. DESIGN: A longitudinal study of motor vehicle accident victims one month and 18 months after the accident. SUBJECTS: Twenty-four motor vehicle accident victims admitted by the trauma team at the Royal Adelaide Hospital. A 52% response rate was achieved. MAIN OUTCOME MEASURES: Post-traumatic stress disorder as diagnosed by the Diagnostic Interview Schedule and disability as measured with the Sickness Impact Profile. RESULTS: Eighteen months after their accidents, six of the 24 subjects had clinically significant PTSD and one was considered borderline. None had been previously diagnosed or treated. The group with PTSD had higher scores on all measures of psychological distress one month after the accident and were more likely to use immature psychological defences. There was no association between physical outcome (measured with the modified Glasgow Outcome Scale) at six months and subsequent diagnosis of PTSD. However, the group with PTSD had higher levels of disability on assessment with the Sickness Impact Profile, particularly in the domain of social functioning. The results suggest PTSD was associated with work-related dysfunction equal to that associated with severe physical handicap. CONCLUSION: The data from this pilot study suggest that PTSD after motor vehicle accidents is an important cause of disability, which may also become the focus for damages in litigation. Thus, there is a need for further investigation of the early patterns of distress and to design preventive programs for victims of road accidents. PMID- 8412953 TI - Excimer laser photorefractive keratectomy: 12 months' follow-up. AB - OBJECTIVE: To report the results of 12 months' follow-up of patients with myopia treated with excimer laser photorefractive keratectomy. DESIGN: A prospective study of patients undergoing excimer laser photorefractive keratectomy for myopia was commenced in September 1991. The study included 50 eyes (38 patients) with preoperative myopia of -1.50 to -6.00 dioptres and a maximum astigmatism of 0.75 dioptres. RESULTS: Forty-six (92%) of the 50 eyes had an uncorrected acuity of 6/6 or better at 12 months, and 49 eyes (98%) had 6/12 or better. No patients had lost their best corrected acuity at 12 months. Forty-five eyes (90%) were within 0.50 dioptres of emmetropia and 47 eyes (94%) were within 1.00 dioptre of emmetropia. CONCLUSIONS: Excimer laser photorefractive keratectomy is a safe and predictable procedure for low to moderate myopia. It will increasingly be offered to patients as an alternative to the use of spectacles and contact lenses. PMID- 8412954 TI - Improving influenza vaccine coverage in at-risk groups. Good intentions are not enough. AB - OBJECTIVES: To raise awareness of the need for annual influenza vaccination in the elderly (those 65 years of age and older), and of the efficacy of organisational practice strategies (specifically at-risk registers and reminder systems) to improve influenza vaccine coverage. DATA SOURCES: Extensive search of the existing literature using MEDLINE and manual searching techniques. Recent key review articles on organisational strategies were also studied. STUDY SELECTION: Primary source articles were included if they evaluated the efficacy of vaccination and the various strategies used to improve coverage. More attention was paid to randomised controlled trials, although other study designs were considered. A formal algorithm for estimating the effect of organisational strategies was not used; rather, appropriate studies which highlighted advantages and disadvantages were selected. RESULTS: Current levels of vaccination for influenza are low. Negative patient attitudes and lack of an organised and systematic approach are the major barriers to improving vaccination rates. Vaccination coverage can be improved by 10%-30% by use of at-risk registers and reminder systems. CONCLUSIONS: Further research is required to determine the impact of both patient attitudes and reminder systems on influenza vaccination rates. A randomised controlled trial is currently underway to look at both these issues. Provider and patient reminder systems should be incorporated into general practice to improve the influenza vaccination rate. PMID- 8412955 TI - The house of Glass: studying occupational health in New Zealand. PMID- 8412956 TI - HIV infection and AIDS in the tropics. AB - One decade after the first description of the acquired immunodeficiency syndrome (AIDS), the epidemic has become a world-wide public health problem. Infection with the human immunodeficiency virus (HIV) is now spreading most rapidly among the poorer populations in the industrialised world and in the developing world. It has become the most important cause of death among adults in the major African cities and it absorbs an ever increasing proportion of health care budgets. On a global scale heterosexual intercourse is now the most common mode of transmission of HIV, resulting in a growing problem of transmission of the virus from mother to child. As a result of the AIDS epidemic, the incidence of tuberculosis is rising in virtually all populations. Developing countries face a double, gigantic challenge in an unfavourable economic and political climate: caring for those infected with HIV and preventing new infections. PMID- 8412957 TI - Surgery in the tropics. AB - The practice of surgery in the tropics provides experience in the management of a broad range of conditions without many of the aids now regarded as essential in developed countries. Clinical judgement must play a greater role but the often advanced stage of disease at presentation may lessen the need for investigations and limit the treatment options. Subspecialisation is well advanced in many countries but most of the surgery performed in the world is by people not long out of medical school, much less by general surgeons. Western trained surgeons need to take into account local cultural attitudes to disease and disease management. Busy practical surgeons in tropical areas have made notable contributions to knowledge and practice. Burkitt and Huckstep, both in Kampala, and Brand in Vellore are outstanding examples. Each of these men was heavily dependent on a great many comparatively untrained but skilled paramedical workers. PMID- 8412958 TI - Anaesthesia. PMID- 8412959 TI - A case of acute hepatitis E in Victoria. PMID- 8412960 TI - Hepatitis B infection in Victoria 1992: time to review the high-risk vaccination strategy. PMID- 8412961 TI - A traveller returning from Nepal with hepatitis E. PMID- 8412962 TI - No role for short term tuberculosis chemoprophylaxis for travellers. PMID- 8412963 TI - Proof of immunisation status required for school entry. PMID- 8412964 TI - Cancer in pregnancy in Queensland. PMID- 8412965 TI - The 18 to 20 week obstetrical scan. PMID- 8412966 TI - Who will deliver the next generation? PMID- 8412967 TI - Paradoxical bronchoconstriction in patients with stable asthma. PMID- 8412968 TI - The cost of an after-hours visit. PMID- 8412969 TI - Late rupture of a mycotic cerebral aneurysm after mitral valve replacement for bacterial endocarditis in the inactive stage. PMID- 8412971 TI - High stakes for public health. PMID- 8412970 TI - Occupational exposure to glutaraldehyde associated with tachycardia and palpitations. PMID- 8412972 TI - Medical journals and the popular media. PMID- 8412973 TI - The doctor as expert witness. PMID- 8412974 TI - [Campaign against epilepsy]. PMID- 8412975 TI - [Hemorrhoids and other proctologic diseases]. PMID- 8412976 TI - [Nutrition in gout]. PMID- 8412977 TI - [Psychiatric ABC for the pharmacist. Part 3]. PMID- 8412979 TI - Corneal surgery for myopia. PMID- 8412978 TI - [Parkinson disease: transplantation of embryonic brain tissue]. PMID- 8412980 TI - Masoprocol for multiple actinic keratoses. PMID- 8412981 TI - Influenza vaccine: an additional note. PMID- 8412982 TI - Drugs for tuberculosis. PMID- 8412983 TI - Interactions between tumoral MCF7 cells and fibroblasts on matrigel and purified laminin. AB - A reconstituted basement membrane (matrigel) and/or fibroblasts promote the growth of human breast tumors in athymic nude mice. We have investigated in vitro the effect of matrigel or purified glycoproteins (laminin and fibronectin) on tumoral MCF7 cells-fibroblasts interactions. In coculture on matrigel, MCF7 cells organized into clusters attached on top of fibroblasts aggregates. During the process resulting in tumor cells-fibroblasts aggregation, fibroblasts actively migrated while MCF7 cells were passively transported. Using purified proteins, specific antibodies and synthetic peptides, we show that cell aggregation induced by immobilized and soluble laminin is antagonized by exogenous fibronectin or fibronectin synthesized by fibroblasts. PMID- 8412984 TI - Extracellular matrix deposition in cultured dermal fibroblasts from four probands affected by osteogenesis imperfecta. AB - Type I procollagen biosynthesis and matrix deposition were studied in cultured fibroblasts of four probands affected by Osteogenesis Imperfecta and in whom the mutations have been characterized. The mutations along the triple helix altered all biochemical parameters considered, i.e. thermal stability, kinetics of procollagen secretion and rate of maturation from procollagen to collagen. The biochemical findings were peculiar for each case considered, but there was no correlation between biochemical parameters and clinical phenotype. In all our probands, regardless of the clinical severity, mutant chains appeared in the insoluble matrix formed by fibroblasts cultured in the presence of dextran sulfate. The densitometric scanning revealed a relative increased amount of fibronectin, suggesting that the matrix contained a lower quantity of type I collagen. Furthermore, the amount of mutant chains found in the insoluble fraction was clearly less than expected, considering that 75% of new synthesized trimers are abnormal. Therefore, in the presence of a mutation, the protein available for extracellular matrix formation is reduced and the mutant trimers incorporated in the matrix probably interfere with normal fibril performance. The abnormal fibril morphology has a dramatic effect in bone, interfering presumably with a correct mineral deposition and interactions with non/collagenous bone proteins. PMID- 8412985 TI - Type I and type III collagen mRNA levels in kidney regions of old and young rats. AB - Interstitial fibrosis is a common feature of renal aging. The steady-state levels of type I and type III collagen mRNAs as well as DNA, protein and collagen deposition were investigated in the cortex, inner and outer medulla of aged (22 months old) rats in comparison to young (5 months old) controls. Our data show that the cortex and outer medulla of old rats expressed significantly higher percentage of type I collagen mRNA compared to the respective regions in the young rat kidneys. Moreover, within the group of the old rats, the cortex expressed significantly higher percentage of type I collagen mRNA compared to the inner medulla whereas in the group of the young rats the expression was similar in all kidney regions. The ratio of extracellular collagen to DNA was significantly higher in the cortex, inner and outer medulla of old compared to young rats. The ratio of collagen to total protein, although showing a similar age-related difference, attained statistical significance in the cortex only. Thus, the present study indicates a close relationship between the expression of the mRNA for type I collagen, the major structural constituent of fibrotic tissues, and the deposition of collagen in both the cortex and outer medulla of the kidney. Moreover, the clear differences found between old and young rat kidneys can serve as markers for renal aging and might explain at least some of the kidney impairments caused by fibrosis during senescence. PMID- 8412986 TI - Atherosclerosis in young white males: arterial collagen and cholesterol. AB - As part of a multicenter study on atherosclerosis, we examined defined segments of thoracic and abdominal aortas from 118 white males, age 15-34 years, who died from external causes. One half of each aorta specimen was graded for lesions. Intima-media preparations were assayed for collagen and cholesterol in two standardized regions (dorsal and ventral) derived from the alternate half of each segment. Even though the mean extent of intimal surface involvement with raised lesions remained minimal (0-6%), the data revealed a remarkable transition in vessel wall chemistry over this time span. For example, the amount of collagen per unit surface area increases with age in all vessel segments except the ventral domain of the thoracic aorta. The amount of collagen as a percent of total vessel protein rises with age only in the ventral and dorsal regions of the abdominal aorta. Free and esterified cholesterol levels per unit surface area increase with age in all vessel segments. There is a significant correlation between collagen and esterified cholesterol per unit surface area in all vessel regions with the exception of the abdominal ventral segment. In the latter segment increases in collagen per unit surface area occur without a corresponding increase in cholesterol level suggesting that connective tissue proliferation may actually precede lipid deposition in the genesis of atherosclerosis. Esterified cholesterol is present at higher levels in the dorsal domains of the thoracic and abdominal aortas than in the ventral domains. These findings provide chemical data confirming that the dorsal domains is the most lesion-prone region of these vessel segments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8412987 TI - Type A modules: interacting domains found in several non-fibrillar collagens and in other extracellular matrix proteins. AB - A 200-amino acid long motif first recognized in von Willebrand Factor (type A module) has been found in components of the extracellular matrix, hemostasis, cellular adhesion, and immune defense mechanisms. At present the extracellular matrix is the predominant site of expression of type A modules since at least four non-fibrillar collagens and two non-collagenous proteins contain a variable number of modules ranging from one to twelve. The modules conform to a consensus motif made of short conserved subregions separated by stretches of variable length. The proteins that incorporate type A modules participate in numerous biological events such as cell adhesion, migration, homing, pattern formation, and signal transduction after interaction with a large array of ligands. PMID- 8412988 TI - Valyl-alanyl-prolyl-glycine (VAPG) serves as a quantitative marker for human elastins. AB - Thermolysin digests of human elastins were examined for reliable elastin peptide markers as determined by HPLC followed by amino acid sequencing of promising peaks. The tetrapeptide VAPG was found to occur in the early portion of the chromatogram in a highly reliable fashion. The peptide appears to be significantly amplified, when compared with the other peptides, in that it is derived from the hexapeptide repeat in elastin, VGVAPG, which repeats itself in two three-piece segments in the c-terminal portion of the tropoelastin molecule. VAPG serves as a highly reliable quantitative measure for human elastins, allowing sensitivities to less than a microgram. Thus, it is a significantly more accurate measure than other existing methods. Precision also appears to be enhanced because of the directness of the measurement. The use of VAPG as a quantitative marker for human elastin has clinical application in the study of elastin-based connective tissue diseases. PMID- 8412989 TI - Hyaluronan synthesis by rat liver stellate cells is enhanced under endotoxic conditions. AB - As part of an investigation of the effect of sepsis on the sinusoidal cells of liver we studied the influence of conditioned media from Kupffer cells (KC) and liver endothelial cells (LEC) from normal rats and animals pretreated with endotoxin (ET) on the hyaluronan (HA) production by stellate cells (SC) in culture. SC proliferation, as measured by direct cell counting and [3H]thymidine incorporation, was also recorded. An SC from an ET-treated rat produces similar amounts of HA as a normal SC when grown in culture medium containing 10% fetal calf serum (FCS). When animals were treated with ET, there was, however, an increase in the yield of isolated SC and thus the whole cell population in ET treated rats has a potential to produce a larger amount of the polysaccharide. Factors present particularly in the KC conditioned media supplemented with 10% FCS from normal and endotoxemic animals greatly stimulated the proliferation and synthesis of HA in SC cultures, after a lag-phase which was four days for normal SC and three days from ET-treated animals. The rate of HA synthesis was closely related to the increase in the amount of thymidine incorporation into DNA. Conditioned media containing 10% FCS obtained from LEC cultures were also stimulatory but were less effective in inducing cell proliferation and production of HA. We suggest that the elevation of plasma HA found in severe endotoxemia and sepsis may be caused in part by the: (i) increased number of SC; (ii) an in vivo activation of SC population which predisposes for HA-production; and, (iii) action of factors or mediators from cells of the reticuloendothelial system of the liver on the cell proliferation and on the HA synthesis in the SC population. PMID- 8412990 TI - A mouse 3T6 fibroblast cell culture model for the study of normal and protein engineered collagen synthesis and deposition into the extracellular matrix. AB - Mouse 3T6 fibroblasts deposited an organized collagenous extracellular matrix during long-term culture in the presence of ascorbic acid. The matrix produced by the cells had a similar distribution of collagen types as the mouse dermal matrix, comprising predominantly type I with smaller amounts of types III and V collagens. By day 8 of culture more than 70% of the collagen in the 3T6 matrix was involved in covalent crosslinkages and required pepsin digestion for extraction. Incorporation of NaB3H4 into reducible crosslinks and aldehydes directly demonstrated the involvement of the alpha 1 (I)CB6 and alpha 2(I)CB3.5 in crosslinks. The pattern of reducible crosslinks in the in vitro 3T6 matrix was similar to that in mouse skin suggesting a comparable fibril organization. Processing of procollagen to collagen occurred efficiently throughout the culture period and the rate of collagen production was unaltered during 15 days of culture, indicating that the development of a collagenous matrix does not directly play a role in procollagen processing or biosynthetic regulation. The existence of a preformed matrix did however, increase the efficiency with which newly synthesised collagen was incorporated into the pericellular matrix. At day 0, when there was no measurable matrix present, 29% of the collagen synthesised was deposited, while by day 15, 88% of the collagen was laid down in the matrix. The development of this 3T6 culture system, where collagen is efficiently incorporated into an organized extracellular matrix, will facilitate detailed studies on matrix organization and regulation and provide a system in which protein-engineered mutant collagens can be expressed to determine their effects on the production of a functional extracellular matrix. PMID- 8412991 TI - A major non-collagenous 62 kDa protein from rat bone mineralized matrix is identical to pp63 a phosphorylated glycoprotein from liver. AB - A protein present as a M(r) 62 k monomer and as several differently sized disulfide-bonded oligomers has been isolated from rat bone mineralized matrix. Its overall tissue distribution determined by ELISA immunoassays showed the protein present only in bone, tooth and in serum while aorta, cartilage, intestine, kidney, liver, muscle, skin, spleen and tendon were all negative. Despite that the 62 kDa protein was abundant and selectively found in bone, no positive cDNA clone could be identified in several rat bone libraries. Positive clones were, however, identified in a rat liver expression library. A cDNA clone of 1.3 kb hybridized in a Northern blotting assay to a 1.8 kb mRNA in rat liver. No hybridization signal was detected with RNA from bone, brain, lung, muscle, spleen and kidney. Sequence analysis of the isolated cDNA clone revealed a 50-bp untranslated region followed by an open reading frame of 357 amino acids. The open reading frame can be divided into a 17-amino acid signal peptide followed by the mature protein of 340 amino acids with alanine as its N-terminal amino acid. A short N-terminal amino acid sequence from the isolated 62-kDa bone protein verified the molecular identity of the cDNA clone. The primary structure of the 62-kDa liver protein was identical to a that of a 63-kDa phosphorylated glycoprotein (pp63) from liver. PMID- 8412992 TI - Neuroblastoma with N-myc amplification detected by urine: mass screening in infants after the sixth month of life. PMID- 8412993 TI - SIOP Working Committee on Psychosocial Issues in Pediatric Oncology. AB - Recognizing the importance of psychosocial issues in the care and cure of the child with cancer, the board of the International Society of Pediatric Oncology (SIOP) in 1991 constituted a Working Committee on Psychosocial Issues in Pediatric Oncology, with Giuseppe Masera as chair and John Spinetta as co-chair. This committee met for the first time in Rhodes, Greece, in October 1991. The committee discussed various psychosocial issues and developed a document on Aims and Recommendations, summarizing the experiences of major centers. This document was approved by the SIOP board, which recommended diffusion of the document to the pediatric oncology community. PMID- 8412994 TI - Ectopic neuroblastoma of the thoracic wall: a case report. AB - A case of thoracic neuroblastoma arising at the anterior mammillary line from the inner surface of the second and third right rib in a 17-month-old male child is reported. Primary surgery allowed excision of the mass. According to the histologic, histochemical, and molecular features of the tumor, all consistent with the diagnosis of neuroblastoma stage 2, the patient underwent adjuvant chemotherapy and, after 2 years, he is alive and disease free at follow-up. In view of different clinical and therapeutical approaches, only a strict adherence to histopathologic criteria allows differentiation of neuroblastoma, even in a very unusual anatomical site, from other small-cell, peripheral neoplasms presumably arising from the embryonic neural crest, such as Askin's tumor, neuroepithelioma, and ectomesenchymoma. The site of origin of these tumors is an important diagnostic clue but can be misleading: only careful microscopic evaluation and molecular analysis lead to incontrovertible diagnosis and yield correct clinical management. PMID- 8412995 TI - Primitive malignant nonepithelial hepatic tumors in children. AB - Primary intrahepatic malignant nonepithelial tumors are very rare in children and account for 2% of all malignant mesenchymatous tumors under the age of 15 years. Clinical presentation, radiologic features, and histologic types are not unequivocal. The predominant role of surgery takes place either initially in small localized tumors or later, after initial reductive chemotherapy. In all cases, complete resection is the necessary but not sufficient condition for cure. Additional radiotherapy seems ineffective. High-dose chemotherapy and/or liver transplantation can be proposed for resistant cases. The disease-free survival rate is 37% at 2 years for the whole series. PMID- 8412996 TI - Incidence and etiology of ifosfamide nephrotoxicity: report of a meeting held in Rhodes, Greece, October 3, 1991, sponsored by Asta Medica, Frankfurt, Germany. AB - The nephrotoxic potential of ifosfamide is now clearly recognized but the true incidence is unknown and risk factors are uncertain. There are, as yet, few studies which systemically explore these issues although many centers have collected data from patients receiving ifosfamide. These support the need for collaborative studies to look at the influence of probable risk factors such as age, cumulative/dose, schedule, and exposure to other nephrotoxic drugs. The ability to detect acute subclinical changes in renal function may provide the opportunity to predict subsequent clinical toxicity. The consensus of opinion recorded so far provides the basis of recommendations for future studies. PMID- 8412997 TI - Adult type (nonembryonal) soft tissue sarcomas in childhood. AB - Nonembryonal soft tissue sarcomas are rare in childhood. We report our experience from a series of 28 patients treated at this centre, whose treatment was based on primary surgical resection. No patient required limb amputation. Anatomical distribution and histological subtypes differed from the adult sarcoma population. A total of 79 percent of the sarcomas were classified as high grade. Six patients (21%) developed local recurrence, and 6 patients (21%) developed distant recurrence, including 3 patients (11%) who developed both. Median follow up was 57 months. Actuarial 5 years survival was 82%, which compares favorably with the 5 year survival of adults with high grade soft tissue sarcomas treated at this centre (63%) and with the survival of children with embryonal sarcomas treated by chemotherapy. PMID- 8412998 TI - Anterior pituitary function and computed tomography/magnetic resonance imaging in patients with Langerhans cell histiocytosis and diabetes insipidus. AB - In order to document anterior pituitary dysfunction in patients with biopsy proven Langerhans cell histiocytosis (LCH) and diabetes insipidus and to correlate this with structural changes on imaging, we performed an insulin tolerance test, enhanced computed tomography (CT), and unenhanced magnetic resonance imaging (MRI) in nine patients. Six of the nine patients had growth hormone deficiency, which in two patients was part of panhypopituitarism and in one was associated with poor cortisol response to insulin hypoglycemia. One patient had an exaggerated growth hormone response and one who had had neck irradiation as an infant, had a high resting thyroid stimulating hormone (TSH) suggesting compensated primary hypothyroidism. All enhanced CTs were abnormal, bony defects being the only abnormality in two patients and opaque mastoids in one. The remaining six patients all had structural changes in the hypothalamic/pituitary region. Unenhanced MRI confirmed the CT findings except in one child who had been treated with radiotherapy in the intervening period, but, in addition, confirmed diabetes insipidus by showing absence of the posterior pituitary bright signal and picked up white matter changes in a child with clinical neurological dysfunction. Our findings indicate that the development of diabetes insipidus in LCH is commonly associated with anterior pituitary dysfunction and is usually associated with structural changes in the hypothalamic/pituitary axis. PMID- 8412999 TI - Avascular femoral head necrosis in pediatric cancer patients. AB - Avascular femoral head necrosis was identified in 15 patients treated at a major pediatric oncology center from 1974 to 1991. The predominant underlying diagnosis was acute leukemia (lymphoblastic, n = 6; non-lymphoblastic, n = 2); two patients had chronic myeloid leukemia, two Hodgkin's disease, and three other solid tumors. Patients ranged from 7 to 27 years of age at diagnosis of this complication, with a median interval of 25 months (range, 0-11 years) from primary diagnosis. Both steroids and radiation therapy appear implicated in the pathogenesis of avascular necrosis: nine patients had received high cumulative doses of prednisone (3.4-14 g/m2), four had received 35-64.8 Gy local irradiation involving the femoral head, and one underwent total body irradiation (12 Gy). Of the 11 surviving patients, six are asymptomatic and fully active. Two patients have joint pain that is responsive to conservative measures. Severe pain and joint deterioration necessitated arthroplasty in two cases, and a third child is undergoing orthopedic evaluation because of worsening symptoms. Thus, like adult cancer patients, children who receive high doses of steroids or local irradiation involving femoral heads are at risk for avascular necrosis. In patients with chronic myeloid leukemia, the complication may be disease related. Although outcomes in our series are better than those reported in adults, the long period of risk for these children and young adults precludes definitive conclusions. PMID- 8413000 TI - Favorable response of pediatric AIDS-related Burkitt's lymphoma treated by aggressive chemotherapy. AB - We describe 4 male children infected by the human immunodeficiency virus (HIV) who developed Burkitt's lymphoma during their disease. The clinical picture was characterized by an insidious appearance of symptoms. All the children suffered for several months from abdominal discomfort and a gradual elevation of their blood lactic dehydrogenase (LDH) level prior to diagnosis. Bone marrow involvement was found in 2 of the patients and jaw involvement in the other 2. After confirming the diagnosis of Burkitt's lymphoma, they were treated according to conventional protocols, with no need to reduce the dose intensity. They all went into complete remission and did not suffer from major opportunistic infections during chemotherapy. None of them relapsed. Two patients died from opportunistic infections 1 and 3 years after diagnosis. The other 2 are alive, 7 years and 6 months after diagnosis. The various characteristics of this unique pediatric group are described and the comparison of the clinical picture in adults is made, together with a review of the relevant literature. PMID- 8413001 TI - Anaplastic large cell lymphoma in childhood. PMID- 8413002 TI - Hemihypertrophy, bilateral Wilms' tumor, and clear-cell adenocarcinoma of the uterine cervix in a young girl. AB - We report the case of a young girl who had hemihypertrophy and developed Wilms' tumor in both kidneys and clear-cell adenocarcinoma of the uterine cervix with no maternal history of stilbestrol ingestion during pregnancy. Karyotype on the second Wilms' tumor showed monosomy 22. The link between hemihypertrophy and Wilms' tumor is well known, but their association with clear-cell carcinoma of the cervix has not been previously described. PMID- 8413003 TI - Hypertrophic osteoarthropathy in childhood malignancy. AB - Hypertrophic osteoarthropathy (HOA), well known in adults, is rarely encountered in children. The clinical features include clubbing of the fingers and toes, arthritis, and a sometimes painful ossifying periostitis of the tubular bones. Apart from a hereditary form (primary HOA), most of the cases encountered in children are secondary and associated with conditions such as chronic suppurative lung processes (e.g., cystic fibrosis), congenital heart disease, biliary atresia, and polyposis coli. The association with malignant disorders, which is relatively common in adults, is very rare in children. In 1986 the authors published a case report of a patient with carcinoma of the nasopharynx who developed HOA. Another similar patient has been encountered. In both, the appearance of HOA was associated with a very poor prognosis. A meticulous research of the literature from 1890 to 1990 revealed only 24 children (19 boys, 5 girls) under the age of 18, with malignancy and associated HOA. Among them were 10 patients with a carcinoma of the nasopharynx, 8 with osteosarcoma, 3 with Hodgkin's lymphoma, 1 with a periosteal sarcoma, 1 with mesothelioma of the pleura, and 1 with carcinoma of the thymus. In five patients with HOA, there were no abnormalities of the lungs, mediastinum, or pleura, and none developed during the course of the disease. Many authors mention the predictive value of HOA, especially in association with malignant tumors. In contrast to suppurative processes in the lungs, in those with neoplastic disease involving the chest, HOA may precede pulmonary symptoms by 1-18 months. A striking feature of HOA in these instances is the reversibility of the complaints after successful treatment of the disorder of the chest, both in benign and malignant conditions. The present case is the second reported by the authors and the first description of a girl with carcinoma of the nasopharynx developing HOA. PMID- 8413004 TI - Carboplatin-induced regression of an optic pathway tumor in a child with neurofibromatosis. AB - Optic pathway tumors are common in children with neurofibromatosis-1 (NF-1). The optimal management of these tumors is unknown, particularly when the optic chiasm and other brain structures are involved. We report the dramatic response to carboplatin in a 10-year-old girl with NF-1 and a progressive optic pathway tumor. Tumor shrinkage was accompanied by striking improvement in visual fields, return of color discrimination, and marked improvement in visual acuity. No significant toxicity was observed. One year following completion of chemotherapy the glioma remains as small or smaller than it was at the conclusion of therapy, and there has been no deterioration of vision. Carboplatin is a promising agent for the treatment of optic pathway tumors in children with NF-1. PMID- 8413005 TI - Non-midline endodermal sinus tumor in the head and neck region: a case report. AB - Germ cell tumors, in particular teratomas, are some of the most commonly found tumors in childhood. Six percent of all germ cell tumors are located in the head and neck region. Endodermal sinus tumors (yolk sac tumors) of the head and neck, exclusive of the central nervous system, are rare. This study reports a 20-month old girl with a mass of 5 x 5 cm on the left temporal area and with bone destruction on CT. The histopathological examination of the excised mass revealed an endodermal sinus tumor. The serum AFP and the LDH levels were elevated at diagnosis. The patient was treated by the BEP protocol (bleomycin, etoposide, and cisplatin), six cycles every three weeks. Partial response was attained after the first and complete response after the fourth cycle. The patient, who is being followed up, has been in remission for five months. PMID- 8413006 TI - Adrenocorticotropic hormone-secreting ganglioneuroblastoma associated with opsomyoclonic encephalopathy: a case report with immunohistochemical study. AB - This report describes a case of a 3-year-old female with Cushing's syndrome associated with ganglioneuroblastoma and opsomyoclonic encephalopathy. Immunohistochemistry showed the tumor to be adrenocorticotropic hormone secreting. At autopsy, a cerebellar degenerative lesion was also demonstrated. PMID- 8413007 TI - A sensitive noninvasive analysis of Pt in external tissues. Followup of Pt deposition following cisplatin treatment. AB - Noninvasive analysis of heavy elements in external tissues by diagnostic-x-ray spectrometry (DXS) is presented. Pt can be detected accurately with sensitivity below 1 microgram/g wet weight of tissue. In the present paper the possibility to monitor Pt accumulation and clearance in the external tissues of cancer patients treated with cisplatin [Cis-diamminedichloroplatinum (II)-cDDP] chemotherapy is reported. The DXS method is based on x-ray fluorescence analysis. Heavy elements in the small skin area of interest are analyzed by their excitation with a monochromatic soft x-ray beam of 14.6 KeV. Spectral L lines of heavy metals such as Pt are detected with minimum interference by other elements in the tissues. Skin Pt levels up to about 6 micrograms/g were observed following several courses of cDDP treatment. The Pt seemed to be homogeneously distributed in different skin areas with similar levels in the dermis and epidermis. The rate of clearance of Pt from the skin (50% in about 30 days) was slower by three orders of magnitude than its clearance from plasma. Further studies may use DXS to establish the accurate kinetics of Pt deposition and clearance in tissues of cDDP treated patients, as well as the exact relation between tissue Pt levels and the development of the drug related late complications. PMID- 8413008 TI - Image information transfer properties of x-ray intensifying screens in the energy range from 17 to 320 keV. AB - The image information transfer properties of a number of x-ray fluorescent screens have been measured for x-ray energies from 17 to 320 keV. The detective quantum efficiency of the screens at each x-ray energy has been determined by separate measurements of the x-ray absorption efficiency and the statistical factor associated with the emission of optical photons upon absorption of an incident x-ray. Data have been recorded for both rare-earth phosphor screens and calcium tungstate screens. The value of the statistical factor for optical photon emission tends toward a constant value as the incident energy increases. Comparisons of the image information transfer properties are presented for several screens, which have been measured over a ten year interval. The utility of the screens for high-energy radiography is discussed. PMID- 8413009 TI - Effects of x-ray tube-voltage ripple and current mode on patient dose. AB - This paper proposes a radiographic sensitivity attenuation curve for screen-films systems. This curve expresses the relative inverse value of mAs value needed to give the same photographic density. This curve decreases more slowly with increasing object thickness than the exposure attenuation curve generally used hitherto. By using the radiographic sensitivity attenuation curve and the exposure attenuation curve, the voltage-ripple dependence of mAs values and patient dose for the same photographic density was obtained. As predicted theoretically in a previous paper [Med. Phys. 18, 921 (1991)], it was verified quantitatively the reverse order that larger ripple values resulted in lower patient dose and lower mAs value for the unsaturating current mode at large ripple. This reverse order clearly appeared in two-pulse units. These phenomena were experimentally verified. As a result it was found that in radiographing with a two-pulse unit, larger mA value results in lower patient dose. PMID- 8413010 TI - Determination of x-ray tube focal spot position. AB - In x-ray spectral measurement using a semiconductor detector system, accurate positioning of the detector in front of the anode is very important. This needs accurate determination of the focal spot position on the target in an x-ray tube. We present a simple technique to determine the focal spot position accurately. Using a pinhole technique, the focal image made by x rays is overlapped on the target image made by visible rays emitted when filament heating takes places. Both images are made in the same geometry with the tube diaphragm and filter removed. PMID- 8413011 TI - Review of MR image segmentation techniques using pattern recognition. AB - This paper has reviewed, with somewhat variable coverage, the nine MR image segmentation techniques itemized in Table II. A wide array of approaches have been discussed; each has its merits and drawbacks. We have also given pointers to other approaches not discussed in depth in this review. The methods reviewed fall roughly into four model groups: c-means, maximum likelihood, neural networks, and k-nearest neighbor rules. Both supervised and unsupervised schemes require human intervention to obtain clinically useful results in MR segmentation. Unsupervised techniques require somewhat less interaction on a per patient/image basis. Maximum likelihood techniques have had some success, but are very susceptible to the choice of training region, which may need to be chosen slice by slice for even one patient. Generally, techniques that must assume an underlying statistical distribution of the data (such as LML and UML) do not appear promising, since tissue regions of interest do not usually obey the distributional tendencies of probability density functions. The most promising supervised techniques reviewed seem to be FF/NN methods that allow hidden layers to be configured as examples are presented to the system. An example of a self configuring network, FF/CC, was also discussed. The relatively simple k-nearest neighbor rule algorithms (hard and fuzzy) have also shown promise in the supervised category. Unsupervised techniques based upon fuzzy c-means clustering algorithms have also shown great promise in MR image segmentation. Several unsupervised connectionist techniques have recently been experimented with on MR images of the brain and have provided promising initial results. A pixel intensity-based edge detection algorithm has recently been used to provide promising segmentations of the brain. This is also an unsupervised technique, older versions of which have been susceptible to oversegmenting the image because of the lack of clear boundaries between tissue types or finding uninteresting boundaries between slightly different types of the same tissue. To conclude, we offer some remarks about improving MR segmentation techniques. The better unsupervised techniques are too slow. Improving speed via parallelization and optimization will improve their competitiveness with, e.g., the k-nn rule, which is the fastest technique covered in this review. Another area for development is dynamic cluster validity. Unsupervised methods need better ways to specify and adjust c, the number of tissue classes found by the algorithm. Initialization is a third important area of research. Many of the schemes listed in Table II are sensitive to good initialization, both in terms of the parameters of the design, as well as operator selection of training data.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8413012 TI - Mechanisms of signal loss in magnetic resonance imaging of stenoses. AB - Some of the factors affecting the signal losses that occur in magnetic resonance images at a stenosis or other region of complex flow have been evaluated. The important determinants of dephasing within a volume element are the net gradient moments, which can be kept small even at long echo times. For compact gradient wave forms, the echo time by itself is unimportant and does not affect signal losses. Reducing the fraction of echo sampled is an alternate method to velocity compensation for reducing gradient moment dephasing that keeps higher moments small. The effects of reducing the fraction of echo sampled on the signal losses in flow distal to a stenosis have been measured experimentally. Another source of signal loss at a stenosis is the variation of the mean phase within a volume element that occurs for flow that varies from one phase encoded view to another. Changes in flow behavior between acquisitions lead to signal displacement in the image. These view to view changes have also been quantified. PMID- 8413013 TI - Two-dimensional pulsatile hemodynamic analysis in the magnetic resonance angiography interpretation of a stenosed carotid arterial bifurcation. AB - A two-dimensional pulsatile hemodynamic analysis based on the finite-element technique was performed on a minimally stenosed carotid artery to identify the possible explanation for the differences in the x-ray and magnetic resonance carotid angiograms of a patient. The magnetic resonance angiogram was obtained by applying the maximum intensity projection algorithm to axial slices, acquired using the time-of-flight principle. The differences in the x-ray and magnetic resonance depictions were interpreted based on velocity profile, wall shear stress, and streamline data provided by the hemodynamic analysis. The specific contribution of the stenosis was further isolated from that of the bifurcation by comparing the flow patterns within the stenotic artery with those of its normal counterpart. The Doppler spectral velocity wave form of the patient constituted the basis for the pulsatile flow velocity specification. The analysis took into consideration the non-Newtonian viscosity of blood. The numerical procedure was validated through different convergence criteria and through shear stress comparisons. The importance of hemodynamic analyses in relation to magnetic resonance angiography was further discussed along with possible shortcomings of the technique. PMID- 8413014 TI - Long term study of random noise and signal uniformity in spin-echo brain imaging. AB - Random noise and MR signal uniformity were analyzed for a period of 9 months using two head coils on a 1.5 T commercial imager. Signal response from a cylindrical phantom filled with a 0.1 mmol/L CuSO4 solution was incorporated in a correction scheme and the effect of correction on uniformity and accuracy of tissue volume determination was measured. There was little change in random noise (CV < 5%) and image uniformity (CV < 15%) over the time of the study. The uniformity in a 130 cm2 region was 4% for a double saddle coil and 5.3% for a mirror coil. After correction, uniformity was improved to 1.1% for the double saddle coil and 1.2% for the mirror coil. In a 40 cm2 central region the uniformity was approximately three times better than in the 130 cm2 region. A second phantom, mimicking the relaxation times of CSF and the brain, consisted of saline encapsulated in vials of volume ranging from 35.6 ml to 249.6 ml and placed in a 0.1 mmol/L solution of MnCl2. Uniformity correction reduced the average error in volume measurement from 8.6% to 6.1%. PMID- 8413015 TI - An unbiased signal-to-noise ratio measure for magnetic resonance images. AB - Estimating the true signal-to-noise ratio (SNR) of magnetic resonance (MR) images with low signal is confounded by the magnitude presentation of the data. This paper suggests a simple solution to this problem. A common method of measuring SNR compares the mean signal to the standard deviation of the noise. This SNR measure was found to be satisfactory for high but not low signal-to-noise image regions because of noise bias. These inconsistencies are removed by introducing unbiased definitions of the signal and noise levels in terms of their root-mean square values. The approaches are compared by evaluating the SNR values for MR medical images. PMID- 8413016 TI - Considerations on the calibration of small thermoluminescent dosimeters used for measurement of beta particle absorbed doses in liquid environments. AB - An investigation has been carried out on the factors which affect the absolute calibration of thermoluminescent dosimeters (TLDs) used in beta particle absorbed dose evaluations. Four effects on light output (LO) were considered: decay of detector sensitivity with time, finite TLD volume, dose linearity, and energy dependence. Most important of these was the decay of LO with time in culture medium, muscle tissue, and gels. This permanent loss of sensitivity was as large as an order of magnitude over a 21-day interval for the nominally 20-microns thick disc-shaped CaSO4(Dy) TLDs in gel. Associated leaching of the dosimeter crystals out of the Teflon matrix was observed using scanning electron microscopy. Large channels leading from the outside environment into the TLDs were identified using SEM images. A possibility of batch dependence of fading was indicated. The second most important effect was the apparent reduction of light output due to finite size and increased specific gravity of the dosimeter (volume effect). We estimated this term by calculations as 10% in standard "mini" rods for beta particles from 90Y, but nearly a factor of 3 for 131I beta particles in the same geometry. No significant nonlinearity of the log (light output) with log (absorbed dose) over the range 0.05-20.00 Gy was discovered. Energy dependence of the LO was found to be not detectable, within measurement errors, over the range of 0.60-6.0 MeV mean energy electrons. With careful understanding of these effects, calibration via gel phantom would appear to be an acceptable strategy for mini TLDs used in beta absorbed dose evaluations in media.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413017 TI - Dosimetry considerations of bone-seeking radionuclides for marrow ablation. AB - Marrow ablation by radionuclide therapy for patients requiring bone marrow transplantation is possible by injecting bone-seeking radiopharmaceuticals. For each radionuclide under consideration, one should determine the (1) amount of activity required to deliver a given radiation dose to marrow, (2) waiting period before reinfusion to limit the radiation dose to the transplant marrow to an acceptable level, and (3) dose to other critical organs. In an attempt to answer these questions, dose estimates for several radionuclides of interest (32P, 90Y, 153Sm, 166Ho, 186Re, and 188Re) have been calculated. All doses are derived following the MIRD methodology. Biodistribution data of 153Sm-labeled phosphates in the rat are used to estimate uptake of similar radiopharmaceuticals in humans. Typically the skeleton retains 50% of the injected activity and 50% is excreted rapidly through the urine, permitting delivery of ablation doses to the marrow, with tolerable doses to other organs. Skeletal retention data measured from a diagnostic dose can be used to calculate the activity required to deliver a desired marrow ablation dose consistent with toxicity limits set by other critical organs. PMID- 8413018 TI - Compensation for attenuation, scatter, and detector response in SPECT reconstruction via iterative FBP methods. AB - Iterative filtered backprojection (FBP) reconstruction is a fast method for single photon emission computed tomography (SPECT) that can provide simultaneous compensation for nonuniform attenuation, scatter, and distance-dependent detector response. The iterative FBP method was applied to the reconstruction of SPECT images of a chest phantom consisting of nonuniform attenuating media. One hundred twenty projections containing 20 million counts were reconstructed on an image array of 128 x 128 x 64. The accuracy of reconstruction was significantly improved, in terms of concentration ratio, object shape, and noise reduction. Results of this experimental study indicate that interactive FBP can effectively reconstruct SPECT images, while other iterative methods, such as maximum a posteriori probability (MAP), will spend at least ten times more computing effort in the reconstruction on the same computer. The disadvantage of iterative FBP is its inability to model the noise properties of data collection accurately, as compared to the MAP methods. PMID- 8413019 TI - Effect of energy resolution on scatter fraction in scintigraphic imaging: Monte Carlo study. AB - The effect of energy resolution in detector systems on scatter fractions in scintigraphic imaging through Monte Carlo simulation is investigated. A 10-cm Tc 99m line source within a cylindrical water phantom 20 cm in diameter and 20 cm in length was modeled and energy spectra were calculated at three different line source positions. The energy resolution was changed from 8% to 16% FWHM at 140 keV and a symmetrical energy window width was varied from 8% to 23% on the photopeak of 140 keV in energy spectra corresponding to each energy resolution. The relationship between the scatter fraction and the symmetrical energy window width, and the relationship between the scatter fraction and the primary counts were presented for all energy resolutions investigated. Furthermore, the effect of the asymmetrical energy window on reducing the scatter fraction was also studied and compared with the narrow symmetrical energy window. The results quantitatively showed that improved energy resolution can considerably decrease the scatter fraction with a narrow symmetrical energy window or an asymmetrical energy window without significant primary count-loss compared to that obtained with lower quality energy resolution. The asymmetrical energy window could reduce scatter fraction as compared with the narrow symmetrical energy window when the same number of primary counts was required for both energy windows. Knowing the relationship between the scatter fraction and the primary counts is important in scintigraphic imaging to select the optimum energy window corresponding to the energy resolution. PMID- 8413020 TI - Dead time of an anger camera in dual-energy-window-acquisition mode. AB - Two side-by-side energy windows are sometimes employed in quantitative single photon emission computed tomography (SPECT) studies. The count-rate losses at high activities for a GE400AT camera were measured in such a dual-window acquisition mode by imaging a decaying source composed of a hot sphere within a warm cylinder. The data in each window was either kept separate or combined for purposes of dead-time correction and the paralyzable model was assumed. In addition, correction factors were derived from a "monitor" source at the edge of the camera. Finally, energy spectra for only the monitor-source region of interest and for the entire camera were obtained under both low- and high-count conditions and compared. With 99mTc (1) the spectral measurements show no peak shift but reveal pulse-pile-up spectral degradation. (2) The monitor-source corrections do not agree well with those from the model, presumably because of the differential effects of such degradation. For this camera the preferred correction method for patients is one using the model and two, effective, phantom derived dead times for the separate data from the two windows. The two effective dead times are needed to compensate for pulse pile-up adding more counts to the lower-energy window than to the higher-energy one at high rates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413021 TI - A vectorized Monte Carlo code for modeling photon transport in SPECT. AB - A vectorized Monte Carlo computer code has been developed for modeling photon transport in single photon emission computed tomography (SPECT). The code models photon transport in a uniform attenuating region and photon detection by a gamma camera. It is adapted from a history-based Monte Carlo code in which photon history data are stored in scalar variables and photon histories are computed sequentially. The vectorized code is written in FORTRAN77 and uses an event-based algorithm in which photon history data are stored in arrays and photon history computations are performed within DO loops. The indices of the DO loops range over the number of photon histories, and these loops may take advantage of the vector processing unit of our Stellar GS1000 computer for pipelined computations. Without the use of the vector processor the event-based code is faster than the history-based code because of numerical optimization performed during conversion to the event-based algorithm. When only the detection of unscattered photons is modeled, the event-based code executes 5.1 times faster with the use of the vector processor than without; when the detection of scattered and unscattered photons is modeled the speed increase is a factor of 2.9. Vectorization is a valuable way to increase the performance of Monte Carlo code for modeling photon transport in SPECT. PMID- 8413022 TI - A theoretical analysis using ratios of the major elements measured by neutron activation analysis to derive total body water, protein, and fat. AB - A theoretical analysis based upon Reference Man is used to derive the water, protein, and fat components of soft tissue using the ratios of carbon/oxygen and nitrogen/hydrogen determined by in vivo neutron activation analysis and measurement of body mineral content by dual energy x-ray absorptiometry. The standard deviations in the estimates of water, protein, and fat are 0.58, 0.59, and 0.53 kg, respectively, if the reproducibility measurement of the carbon/oxygen ratio is 2.5% and of the nitrogen/hydrogen ratio is 5%, the accuracy of measurement of bone mineral is 4%, and estimate of glycogen is 0.5 kg. New neutron activation technology may permit sufficiently precise measurements. PMID- 8413023 TI - A rho-theta technique for treatment verification in radiotherapy and its clinical applications. AB - A new technique based on a rho-theta coordinate system for determining differences in patient position between portal and simulator images is presented. Unlike the conventional point matching method, which requires the fiducial points to be labeled in pairs before the registration, the rho-theta technique avoids this manual procedure. It accomplishes the treatment verification in two major steps; image alignment and field displacement analysis. For the same number of fiducial points in the simulator and portal images, it first finds the corresponding paired points if the points are not distributed symmetrically about their centroid. This is followed by alignment of these paired points using the least squares matching method to find the optimal two-dimensional rigid body transformation parameters (shift, rotation, and scaling factor). The transformation parameters are then used to transform the portal field edge into the simulator image, so that the portal field can be compared with the prescribed field on the simulator image. A number of parameters were explored to describe the field displacement errors, including treatment field size, under/over irradiated size, the shift in center of gravity of the field, the field edge shift, and rotation of the field. The rho-theta technique as implemented is both fast and accurate. Experiments on the registration of radiological phantom portal images acquired with an on-line portal imaging system mounted on a linear accelerator indicate an accuracy on the order of 1 mm in detecting the shift of the field's center of gravity and approximately 1 degree in detecting the field rotation. The results of a clinical trial are also presented.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413024 TI - Automated determination of patient setup errors in radiation therapy using spherical radio-opaque markers. AB - Patient positioning accuracy can be quantified by the three-dimensional (3-D) translations and rotations required to transform the patient back to the desired position. Results of the current study show that the translations and rotations could be obtained from two projection images obtained radiographically on a linear accelerator when spherical radio-opaque markers were implanted inside or affixed to the surface of a skull phantom. In this study, film used to record the images were converted into digital gray scale images using a laser film digitizer. The marker images were located automatically by the computer using image processing techniques. By combining information from both projections, the 3-D locations of the markers were determined to submillimeter precision. Pairs of films were also analyzed for the phantom displaced from its original location by known amounts. The accuracy of the computed translations and rotations required for realignment of the phantom were found to be better than 1 mm and 0.3 degrees, respectively; comparable to the accuracy of the readout system of the equipment used. The general methodology could be coupled with an electronic portal imaging device for use in computer aided or automated correction of patient position in radiotherapy. PMID- 8413025 TI - Computer-assisted positioning of radiotherapy patients using implanted radiopaque fiducials. AB - For certain external beam radiotherapy procedures, precise alignment of patients with the treatment beam is essential for good treatment outcome. A method has been developed for quickly achieving precise patient alignment with the aid of stereoscopically located fiducial markers. The alignment algorithm is developed from standard rigid body mechanics using closed form solutions, obviating the need for iterative fitting methods. The technique is implemented with a digitizing tablet and plane film radiographs. The accuracy of alignment with this method in phantom studies is better than 1 mm and 1 deg relative to CT data. The repeatability of positioning is 0.5 mm (standard deviation) and 0.36 deg (standard deviation). PMID- 8413026 TI - Monte Carlo optimization of metal/phosphor screens at megavoltage energies. AB - The physics of imaging with metal/phosphor (Gd2O2S:Tb on brass) screens at megavoltage energies has been investigated using Monte Carlo simulation. It has been found that pair production is a significant contributor to energy deposition for Bremsstrahlung beams with energies greater than 6 MV. The effects of different thicknesses of phosphor and metal have been studied, and it is shown that the metal plays a significant role in establishing electronic equilibrium in the phosphor. The transport of optical photons through the phosphor has been modeled, and was found that only 10% to 20% of the light created in the phosphor escapes from the surface, with much of the loss being due to total internal reflection at the surface. Calculated results have been compared with experimental measurements of screen brightness for different phosphor and metal thicknesses. The SNR of a video electronic portal imaging device (VEPID) has been calculated as a function of x-ray and optical photon detection efficiency. The non-Poisson distribution of energy deposition in the phosphor is an important contributor to the SNR. The results of this paper should serve as a useful guide to the engineering design of future electronic portal imaging systems. PMID- 8413027 TI - The effect of strong longitudinal magnetic fields on dose deposition from electron and photon beams. AB - The use of strong, uniform, longitudinal magnetic fields for external electron and photon beam irradiation is considered. Using the EGS4 Monte Carlo code modified to account for the presence of magnetic fields, dramatic narrowing of penumbra for photon and electron irradiations is demonstrated. In the vicinity of heterogeneities, "hot" and "cold" spots due to multiple scattering in electron beams are reduced substantially. However, in the presence of strong magnetic fields, the effect of inhomogeneities can be observed far from the location of the inhomogeneity due to reduced "washout" caused by lateral multiple scattering. The enhanced "Bragg peak," proposed or calculated by other authors, is not observed on the central axis of broad beams, owing to lateral equilibrium. It is proven that for broad parallel beams, the central axis depth-dose curve is independent of the strength of the external longitudinal magnetic field, as long as it is uniform. However, strong longitudinal magnetic fields can induce enhancements by redirection of the electron fields coming from point sources. Strong uniform longitudinal magnetic fields provide a way of controlling the spreading of electron beams due to multiple scattering, making the electron beams more "geometrical" in character, simplifying dose-deposition patterns, possibly allowing electron beams to be used in new ways for radiotherapy. Photon therapy also benefits from strong uniform longitudinal magnetic fields since the penumbra or other lateral disequilibrium effects associated with lateral electron transport can be eliminated. PMID- 8413028 TI - Beam quality specification for photon beam dosimetry. AB - It is argued that %dd(10), the percentage depth dose at 10 cm in a 10 x 10 cm2 photon beam at a SSD of 100 cm, is a better beam quality specifier for radiotherapy beams than the commonly used values of TPR10(20) or nominal accelerating potential (NAP). For radiation dosimetry purposes, TPR10(20) is not an ideal beam quality specifier because (i) stopping-power ratios for the same value of TPR10(20) can vary by up to 0.7% for thick-target bremsstrahlung beams; (ii) the value of TPR10(20) becomes insensitive to beam quality changes for high energy beams; and (iii) it has little intuitive meaning. In contrast, %dd(10) in a pure photon beam specifies stopping-power ratios within 0.2% for all thick target bremsstrahlung beams, maintains its sensitivity for high-energy beams, and has a simple physical and clinical meaning. It is shown that for all thick-target bremsstrahlung beams the spr (water/air) = 1.2676-0.002 224[%dd(10)] with a rms deviation of 0.1%. The effects of electron contamination in typical high-energy clinical beams can be corrected for using previously published experimental results or by reducing electron contamination using lead scattering foils. PMID- 8413029 TI - Calculation of absorbed dose ratios using correlated Monte Carlo sampling. AB - A correlated sampling variance reduction technique has been implemented in the EGS4 (Electron Gamma Shower version 4) Monte Carlo code system in order to calculate correction factors for radiation dosimeters. These correction factors are calculated as the ratios of absorbed doses in various geometries irradiated by photon and electron beams. In this paper the efficiency and accuracy of the calculation of the absorbed dose ratios with correlated Monte Carlo sampling is discussed. Theoretical analysis shows that the calculation efficiency and accuracy increases with the degree of correlation between the calculated doses in the individual geometries. Calculations of the absorbed doses in thin slab geometries and in air-filled ionization chambers have been performed. The results show that the gain in computing efficiency is up to a thousand fold for the calculations of the effect of the central electrode material and size on the response of cylindrical ionization chambers. The calculation accuracy (systematic uncertainty) is significantly improved when good correlation exists between the calculated doses in similar thin-slab geometries. PMID- 8413030 TI - Optimization of the dose delivery in a few field techniques using radiobiological objective functions. AB - A method for finding optimal primary fluence profiles for multiple field external beam radiation therapy techniques has been developed using a radiobiologically based objective function that quantifies the probability of achieving complication-free tumor control, P+. The objective function P+ has the valuable property of giving the highest possible dose to the tumor without causing severe damage to normal tissues at risk. This radiobiologically based objective function selects suitable dose levels but also takes into account the dose homogeneity in the target volume to the extent that it is not causing an excessive risk of local recurrence or damage to surrounding normal tissues. The biological parameters used can either be patient specific, as determined by a predictive assay on biopsy specimens, or taken from a library of radiobiological parameter values characteristic for different tissue types of a reference patient. In its present form the method can be used to determine the optimum incident photon fluence profiles for each beam. The method has been used to investigate for a given target volume a large number of combinations of beam entry directions to find the best beam orientations with respect to the probability of achieving complication free tumor control. It is demonstrated that when nonuniform dose delivery is available it is unsuitable to combine parallel opposed beams in two-beam techniques and to a lesser extent also to use perpendicular beams. In two-beam techniques the best angle between the beams is generally in the 100 degrees-120 degrees range. The major symmetry characteristics of the P+ phase space for two beam techniques are also identified. The method can easily be extended from two to three dimensions and noncoplanar geometry, but it is presented here in its two dimensional form for clarity and speed of calculation. PMID- 8413031 TI - A mathematical basis for selection of wedge angle and orientation. AB - The treatment plan optimization criterion that dose be homogeneous over an irradiated volume is equivalent to the criterion that the magnitude of the dose gradient be zero throughout that volume. If the dose gradient due to an individual beam is represented by a vector, the dose gradient due to an ensemble of beams is given by the weighted vector sum of the constituent beams' individual gradients. Given a fixed ensemble of beams, the two ways in which the total dose gradient can be modified are (1) by changes in relative beam weights and/or (2) by changes in the direction and/or magnitude of the dose gradient of one or more of the individual beams. Conventional wedges provide a simple mechanism for altering the dose gradient of a single beam. This paper describes a mathematical basis for the selection of wedge angles, wedge orientations, and relative beam weights, with the goal of producing a field of zero gradient over the volume of beam intersection. The approach is based on 3-dimensional vector analysis of dose gradients, and is valuable not only for its formalism, but also for the conceptual basis it provides for discussing and solving general wedge selection problems. PMID- 8413032 TI - Use of an octree-like geometry for 3-D dose calculations. AB - A representation used for 3-D graphical objects, the "octree," has been applied to the geometrical calculations needed to perform photon beam does calculations for radiotherapy treatment planning. This representation allows the algorithm to attempt to minimize the number of distinct geometrical calculations that are needed to perform dose calculations to a particular resolution. In this way, the calculational time can be minimized, since the geometrical part of the dose calculations is often the most time-intensive part of the calculation process. The octree-like system used here has sped up the photon dose calculations described here by up to a factor of 10. PMID- 8413033 TI - Equivalent squares of irregular photon fields. AB - The tables of equivalent fields published by the British Journal of Radiology (BJR) are intended for calculation of depth-dose functions in rectangular photon fields. We have investigated the validity of the equivalent-field concept for fields of arbitrary shape over a range of photon energies, field sizes and depths. We show that the empirical scatter-radius function (Day function) used to generate the equivalent-field tables is a good approximation to the average over energy of normalized scatter-air ratios extracted from BJR beam data for depths up to 10 cm. However, this function tends to diverge from the data as depth increases. Accuracy can be improved by making the Day function depend on depth. Equivalent squares, determined by sector integration of the original or modified Day functions, are suitable for megavoltage photon-beam dose calculations at central-axis and off-axis points in irregular as well as rectangular fields. PMID- 8413034 TI - Evaluation of electron-beam uniformity during commissioning of a linear accelerator. AB - Electron-beam profiles measured during acceptance testing and commissioning of a linear accelerator were analyzed according to manufacturer's specifications and more stringent AAPM TG-25 recommendations. The difference between specifications of the AAPM report and those experienced commercially is discussed. Although all the electron beams met the manufacturer's specifications, the uniformity parameters for some of them were outside the limits suggested by the AAPM report. Profiles acquired at depths larger than dmax were extremely valuable for evaluation and adjustment of the beams. The following suggestions for practical machine commissioning are made: Profiles taken deeper than at dmax are extremely useful indicators of the beam nonuniformity; acquisition of these profiles should be a part of the routine acceptance testing procedure; and beam uniformity should be evaluated against AAPM recommendations, and this should be included in the purchase specifications. PMID- 8413035 TI - New method for shielding electron beams used for head and neck cancer treatment. AB - Shields and stents of metals with high atomic number, which are custom cast in molds from the melt, are the materials most widely used to protect surrounding tissues during treatment of skin or oral lesions with therapeutic electron beams. An improved fabrication method is to mix a polysiloxane-metal composite, which is readily cast at room temperature by combining a metal-powder/polysiloxane resin mixture with a hardening catalyst. The purpose of the present study is to compare the shielding effectiveness of two different metal-polysiloxane composites with that of conventional cast Lipowitz metal (50.1% Bi, 26.6% Pb, 13.3% Sn, 10% Cd). Also, a 2(3) factorial experiment was run to investigate the effects and interactions of metal particle size (20-microns vs 100-microns diameter), the atomic weight of the metal (304 stainless steel vs 70% Ag, 30% Cu alloy), and the presence or absence of a layer of unfilled polymer added to the forward-scatter side of the shield. The composites of different thicknesses were made by blending 90% (w/w) metal powder separately with 10% polysiloxane base and catalyst. A thin GafChromic dosimeter film was placed between the shielding material and a polystyrene base to measure the radiation shielding effect of composite disc samples irradiated with a 6-MeV electron beam normal to the flat surface of the disc. The results show that composite shields with the metal of higher atomic weight and density (Ag-Cu) combined with an additional unfilled layer are more effective than the stainless-steel composite with a similar additional unfilled layer, in terms of diminishing the dose at the surface of the polystyrene backing material.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413036 TI - An evaluation of the EGS4 and CYLTRAN Monte Carlo codes with regard to boundary beta-ray dosimetry by comparison with experimental beta-ray dose backscatter factors. AB - Beta-ray dose backscatter factors or dose ratios at planar soft-tissue boundaries were calculated using the EGS4/RESTA and CYLTRAN (version 2.1) Monte Carlo codes and these data were compared with experimental results. Since the beta-ray source was 32P, this work addressed the transport of, and energy deposition by, electrons less energetic than 2 MeV. In particular, the simulations targeted the codes' performances with regard to the transport of low energy electrons across material boundaries and the backscattering of low energy electrons. In general, backscatter factors calculated at 7.25 mg/cm2 from several soft-tissue interfaces agreed with experimental values to within about five percent. CYLTRAN was also used to calculate the variation of backscatter factor with distance from aluminum/soft-tissue and air/soft-tissue interfaces and was found to reproduce the shapes of experimental backscatter factor depth profiles. PMID- 8413037 TI - The effect of lead, gold, and silver backings on dose near 125I seeds. AB - Brachytherapy for ocular melanoma uses 125I seeds backed by a gold shield. Conflicting results are reported in the literature on the effect of the gold on dose close to the seeds. In this work, a small lucite jig was constructed such that the seed-to-detector separation remained fixed as high-Z materials of lead, silver, and gold were moved in and out of position behind the seed. The jig was clamped in place in the water filled tank of a beam scanning system. The response of two p-type silicon diodes was measured at several distances from the seed with and without the high-Z backings. The response with the high-Z backing relative to water, found to be the same for each diode and the same for lead and gold, decreased from about 1.01 at 1.5 mm to about 0.92 at 20 mm. It has been suggested in the literature that L-shell fluorescent x rays of approximately 10 keV from the gold backing might contribute significantly to the dose within 7 mm of the seed. To test this, the response with the gold backing relative to water was measured with an aluminum cap of 1-mm wall thickness covering the diode. The cap transmits about 70% of the 125I influence but is essentially infinitely thick to 10-keV photons. The relative response (gold/water) was the same with and without the cap showing that the contribution of 10-keV x rays is negligible. Compared to water, the silver backing was found to enhance the diode response by about 14% between 5 to 10 mm from the seed. PMID- 8413038 TI - The half-life of high dose rate Ir-192 sources. PMID- 8413039 TI - Medical accelerator safety considerations: report of AAPM Radiation Therapy Committee Task Group No. 35. AB - Ensuring safe operation for a medical accelerator is a difficult task. Users must assume more responsibility in using contemporary equipment. Additionally, users must work closely with manufacturers in promoting the safe and effective use of such complex equipment. Complex treatment techniques and treatment modality changeover procedures merit detailed, unambiguous written procedural instruction at the control console. A thorough "hands on" training period after receiving instructions, and before assuming treatment responsibilities, is essential for all technologists. Unambiguous written instructions must also be provided to guide technologists in safe response when equipment malfunctions or exhibits unexpected behavior or after any component has been changed or readjusted. Technologists should be given a written list of the appropriate individuals to consult when unexpected machine behavior occurs. They should be assisted in identifying aberrant behavior of equipment. Many centers already provide this instruction, but others may not. Practiced response and discussion with technologists should be a part of an ongoing quality assurance program. An important aspect of a safety program is the need for continuous vigilance. Table III gives a summary of a comprehensive safety program for medical accelerators. Table IV gives a list of summary recommendations as an example of how one might mitigate the consequences of an equipment failure and improve procedures and operator response in the context of the environment described. Most of these recommendations can be implemented almost immediately at any individual treatment center. PMID- 8413040 TI - An artificial neural network for estimating scatter exposures in portable chest radiography. AB - An adaptive linear element (Adaline) was developed to estimate the two dimensional scatter exposure distribution in digital portable chest radiographs (DPCXR). DPCXRs and quantitative scatter exposure measurements at 64 locations throughout the chest were acquired for ten radiographically normal patients. The Adaline is an artificial neural network which has only a single node and linear thresholding. The Adaline was trained using DPCXR-scatter measurement pairs from five patients. The spatially invariant network would take a portion of the image as its input and estimate the scatter content as output. The trained network was applied to the other five images, and errors were evaluated between estimated and measured scatter values. Performance was compared against a convolution scatter estimation algorithm. The network was evaluated as a function of network size, initial values, and duration of training. Network performance was evaluated qualitatively by the correlation of network weights to physical models, and quantitatively by training and evaluation errors. Using DPCXRs as input, the network learned to describe known scatter exposures accurately (7% error) and estimate scatter in new images (< 8% error) slightly better than convolution methods. Regardless of size and initial shape, all networks adapted into radial exponentials with magnitude of 0.75, perhaps implying an ideal point spread function and average scatter fraction, respectively. To implement scatter compensation, the two-dimensional scatter distribution estimated by the neural network is subtracted from the original DPCXR. PMID- 8413041 TI - Automated selection of regions of interest for quantitative analysis of lung textures in digital chest radiographs. AB - In order to implement a computerized scheme for quantitative analysis of interstitial lung disease in chest radiographs in clinical situations, a fully automated method of selecting many square regions of interest (ROIs) in peripheral lung areas are developed. First, the peripheral lung regions are identified, based on the automated detection of lung apices, ribcage edges, and diaphragm. Then a large number of ROIs are selected sequentially by filling in the identified peripheral regions. Finally, those ROIs containing sharp edges are removed based on an edge gradient analysis, for which a gradient-weighted edge orientation histogram is employed. Approximately 200-400 ROIs were automatically selected for each case with this method. The evaluation of using ROC analysis indicated that the automated ROI selection method was effective in quantitative analysis of lung textures in digital chest radiographs. PMID- 8413042 TI - Computer-aided diagnosis: detection and characterization of hyperparathyroidism in digital hand radiographs. AB - An automated method is developed for the detection and staging of skeletal changes due to hyperparathyroidism on digitized hand radiographs. Subperiosteal bony resorption, particularly along the radial margins of the middle and proximal phalanges, is among the earliest manifestations of secondary hyperparathyroidism. In order to quantify the severity of bone resorption in these regions, the computer method analyzes the roughness of the phalangeal margins, as projected on the radiograph. The regions of interest, which contain the phalanges, are obtained from the digitized hand radiographs by an image preprocessor. The radial margin of each phalanx is detected by a model-guided boundary-tracking scheme. The roughness of these boundaries is then quantified by the mean-square variation and the first moment of the power spectrum. A receiver operating characteristic (ROC) study comparing the computer detection of hyperparathyroidism with the diagnosis by three experienced skeletal radiologists was performed by evaluating 84 hand images from 22 patients. Our present computer method can achieve a true positive rate of 94% and a true-negative rate of 92%. Such a computer-aided diagnosis system may assist radiologists in their assessment of primary and secondary hyperparathyroidism, since it is both accurate and not subject to either intra- or interobserver variations. PMID- 8413043 TI - A novel algorithm for the edge detection and edge enhancement of medical images. AB - A novel algorithm, histogram shifting (HS) is presented, which performs edge detection or edge enhancement with the choice of two parameters. The histogram of a region surrounding each pixel is found and translated toward the origin, resulting in the new pixel value. Images from a variety of medical imaging modalities were processed with HS to perform detection and enhancement of edges. Comparison with results obtained from conventional edge detection (e.g., Sobel) and with conventional edge-enhancement algorithms is discussed. HS appears to perform the edge-detection operation without introducing "double-edge" effects often obtained with conventional edge-detection algorithms. HS also appears to perform edge enhancement without introducing extensive noise artifacts, which may be noticeable with many edge-enhancement algorithms. PMID- 8413044 TI - The need for investigating the optical transfer functions in several field orientations for nonisotropic radiographic systems. AB - In this paper, it is demonstrated that obtaining OTF's for only two directions, parallel and perpendicular to the x-ray tube axis, is insufficient to completely describe the performance of nonisotropic radiographic systems. By performing experiments with three radiological systems in which slit images were obtained for ten different directions, we confirmed that OTF's vary in a nonlinear fashion for directions that are intermediate to the parallel and perpendicular ones. Moreover, we have also identified for each system a range of field orientations- referred to as "Optimum Region"--where sharper images can be obtained. These experimental results can be accounted for by the transfer functions theory. PMID- 8413045 TI - [Patients with type-II diabetes mellitus and neuropathy have no deficiency of vitamins A, E, beta-carotene, B1, B2, B6, B12 and folic acid]. AB - The present study was aimed to determine the vitamin status of vitamins A, E, beta-carotene, B1, B2, B6, B12 and folate in plasma using HPLC and vitamins B1, B2 and B6 in erythrocytes using the apoenzyme stimulation test with the Cobas-Bio analyzer in 29 elderly type II diabetic women with (G1: n = 17, age: 68.6 +/- 3.2 years) and without (G2: n = 12, age: 71.8 +/- 2.7 years) diabetic polyneuropathy. The basic parameters as age, hemoglobin A1c, fructosamine and duration of the disease did not differ in both groups. Furthermore, retinopathy was assessed with fundoscopy and nephropathy with creatinine clearance. The creatinine clearance (G1: 50.6 +/- 3.4 vs. G2: 63.6 +/- 3.7 ml/min, 2p < 0.025) and the percentage of retinopathy (G1: 76.5% vs. G2: 16.7%, 2p = 0.002) were different indicating that G1 had significantly more severe late complications than G2. Current plasma levels of all measured vitamins (A, E, beta-carotene, B1, B2, B6, B12 and folate) and the status of B1, B2 and B6 in erythrocytes did not vary between the two groups (2p > 0.1). In summary, we found a lack of association between the actual vitamin condition in plasma and erythrocytes and diabetic neuropathy. PMID- 8413046 TI - [Serotonin concentration in serum of patients with generalized tendomyopathy (fibromyalgia) and chronic polyarthritis]. AB - The serum concentration of serotonin (S-5-HT) was measured in 31 patients with primary fibromyalgia, 21 patients with rheumatoid arthritis (RA) (15 of them with secondary fibromyalgia) and 20 healthy volunteers. Both patients with primary fibromyalgia and rheumatoid arthritis had significantly lower S-5-HT levels when compared to healthy controls, and S-5-HT concentrations in patients with secondary fibromyalgia were even significantly lower than those of RA-patients. Unlike the patients with rheumatoid arthritis, a significant correlation between S-5-HT level and the number of "tender points" as well as mean pressure tenderness at 24 different points was found in patients with primary fibromyalgia. Conversely, in patients with rheumatoid arthritis the S-5-HT level correlated significantly with erythrocyte sedimentation rate. These results suggest different pathological mechanisms of S-5-HT decrease in patients with primary fibromyalgia and rheumatoid arthritis. On the other hand, they raise the question whether secondary fibromyalgia may be a pathogenetically different syndrome mimicking symptomatically primary fibromyalgia. PMID- 8413047 TI - [Epidemiology of antirheumatic drug-induced ulcer in comparison with peptic gastroduodenal ulcer. Five-year analysis of a hospital patient sample]. AB - Within the framework of a five-year analysis of a hospital population (n = 503), the epidemiological and diagnostic data of patients with non-steroidal anti inflammatory drugs-associated ulcers of the stomach and duodenum (AIU) were evaluated and compared with those of patients with peptic ulcers (PUC) in the stomach and duodenum with no history of NSAIDs use. With respect to age and sex, patients with AIU were significantly older, in particular in the case of gastric ulcer, and revealed a predominance of female sex as compared with those with PU. Episodes of ulcer disease showed no seasonal dependence, either in the AIU or in the PU group. The use of tobacco and alcohol was comparable in both groups. Nor was any difference found between city and rural populations in either group. Laboratory data revealed a higher rate of pathological deviations in the AIU as compared with the PU group, which, however, was unequivocally due to the larger percentage of elderly patients in the former. With respect to blood group distribution, no differences were observed between the groups. An analysis of ulcer site showed a small preponderance of gastric ulcer (59%) over duodenal ulcer (41%) in the AIU group, while the distribution in the case of the PU group was virtually identical (gastric ulcer 51%, duodenal ulcer 49%), with only small differences in site within the stomach and duodenal bulb resp. Analysis of the risk indicators for ulcer bleeding (AIU 66%, PU 50%) revealed the following pattern: in the case of AIU, patients aged over 65, and in the case of AIU and PU, overweight males, bled significantly more often than, respectively, younger patients and female or normal-weight patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413048 TI - [Results of regular follow-up after removal of colorectal adenomas]. AB - After resection of colorectal adenoma 453 patients were included in a follow-up program between 1. 1. 1980 and 30. 6. 1991. 275 patients underwent regular follow up examinations, the primary drop-out rate was 39.2%. In the 275 participating patients 492 primary adenoma were removed. 174 patients (63.3%) had a single adenoma, while 101 patients showed multiple primary adenoma (36.7%). The average follow-up interval was 37 (+/- 28.2) months. During follow-up we found new adenoma in 105 patients (38.2%), the Kaplan-Meier estimation of the five-year recurrence rate was 55.9% (95% confidence-interval 46.9 to 64.8%). The risk of recurrence was increased in patients with multiple primary adenoma (p = 0.003) and multivariate risk analysis showed an independent influence of histology on recurrence rate (p = 0.07). In six patients colorectal carcinoma were detected during adenoma follow-up. During regular follow-up examinations after surgical treatment for colorectal adenoma in more than 30% of all patients new colorectal polyps were detected and removed. Therefore follow-up for colorectal adenoma is effective in prevention of colorectal carcinoma. Still frequency of follow-up examinations have to be adapted to the individual risk of recurrence for each patient. PMID- 8413049 TI - [Unusual course of anti-basement membrane antibody nephritis]. PMID- 8413050 TI - [Prostate cancer: cytogenetic and molecular biology principles]. PMID- 8413051 TI - [Local catheter thrombolysis]. PMID- 8413052 TI - [When must, when should, when can ACE inhibitors be administered in heart failure?]. PMID- 8413053 TI - [Endocarditis caused by Actinobacillus actinomycetemcomitans]. PMID- 8413054 TI - Morphine elimination and hepatic blood flow--a study in patients with portal hypertension. AB - To investigate the influence of changes in portal venous blood flow on the clearance of morphine, the elimination of a single intravenous dose of morphine sulphate was studied in twelve patients with periportal fibrosis undergoing distal splenorenal shunt for portal hypertension. All patients had almost normal preoperative liver function. Six patients (Group A) received morphine after induction of anesthesia but before surgery; a further six patients (Group B) received morphine after completion of the splenorenal shunt and removal of all vascular clamps. Blood was sampled for 24 hours after morphine administration and analysed for plasma morphine concentration using high performance liquid chromatography. The maximum concentration of morphine was significantly greater in those patients in Group B who received morphine after the shunt (P < .002). Clearance of morphine during the operation was also significantly reduced in these patients compared with the initial clearance (before release of the clamps) in Group A (P < .004). As a consequence of these changes the AUC was greater in this group (P < 0.004). However, there were no differences with the elimination rates after surgery had finished. The results suggest that in individuals with normal liver function, the clearance of morphine is dependent on hepatic blood flow. PMID- 8413055 TI - The anticonvulsant effects of propofol, diazepam, and thiopental, against picrotoxin-induced seizure in the rat. AB - The anticonvulsant effects of propofol administered intravenously against picrotoxin-induced seizure were studied and compared with those of diazepam and thiopental in the rat using picrotoxin-seizure threshold. Comparable doses of the three agents ranging between 1.25-20.0 mg/kg, produced dose-dependent increases in picrotoxin-threshold dose. At the lowest administered dose (1.25 mg/kg), there were no significant differences in picrotoxin-threshold dose among the three agents. Using 2.5 mg/kg dose, propofol and diazepam were equally effective and both were significantly more effective than thiopental in increasing picrotoxin threshold dose. At higher doses of anticonvulsants (10.0 and 20.0 mg/kg), propofol was significantly more effective than both diazepam and thiopental. These results indicate that propofol is an effective anticonvulsant against picrotoxin-induced seizure, and this effect is significantly greater than diazepam and thiopental at doses producing clear sedative and behavioral effects. PMID- 8413056 TI - Ketamine anesthesia for short transurethral urologic procedures. AB - Ketamine (K) is a good analgesic and anesthetic agent in short procedures, but the associated cardiovascular responses and emergence reactions limit its use. Benzodiazepines have been used to improve recovery with favourable reports for midazolam (M). Methylphenidate (MPH), the mild CNS stimulant, improves behaviour and mental concentration and can be used to improve recovery from K anesthesia. This was tested, alone and in combination with M by a double-blind study in 30 patients subjected to short transurethral urologic procedures. Patients were randomized into 3 equal groups to receive K-MPH, K-M or K-M-MPH. M (7.5 mg) was mixed with K and MPH (20 mg) was given at the end of urologic procedures. Perioperative monitoring included pulse rate, blood pressure, ECG, and plasma catecholamines. Recovery was assessed by a triad VAS and recovery area was calculated. Distribution-free statistics were used to assess intergroup differences of similar variables. Ketamine produced satisfactory anesthesia for short transurethral urologic procedures. Addition of M did not change the cardiovascular responses of K but resulted in smooth recovery with no changes in the recovery scores. MPH did not improve the recovery scores but increased the incidence of vomiting, excessive talking, and limb movements. PMID- 8413057 TI - Midazolam for prolonged intravenous sedation in patients with tetanus. AB - Midazolam was used for prolonged sedation in ten adult patients with severe tetanus in the Intensive Care Unit for periods ranging from 10 to 50 days. An infusion was started soon after induction and adjusted according to clinical needs. All the patients were intubated and ventilated and in addition received morphine 1-2 mg/hr and alcuronium or pancuronium infusion. Two patients died from complications of tetanus. Six patients recovered from the effects of midazolam within 24 hrs of the cessation of infusion. Recovery took 48 and 72 hrs respectively in the other two patients. Midazolam is a suitable drug for long term sedation in patients with tetanus. However, dose requirements are variable and recovery may be delayed in some patients. PMID- 8413058 TI - Regurgitation following induction of anesthesia in a patient with Zenker's diverticulum--a case report. PMID- 8413059 TI - Epidural morphine analgesia in Qatar. PMID- 8413060 TI - FIO2 of 0.5 in one lung anesthesia. PMID- 8413061 TI - The provision of anesthesia in east Africa--a conundrum. PMID- 8413062 TI - Transtracheal jet ventilation in an adult patient with severe hereditary angioneurotic edema. PMID- 8413064 TI - "Pre-emptive analgesia" a new era of pain control. PMID- 8413063 TI - Hypoglycemia of the diabetic patient during spinal anesthesia. PMID- 8413065 TI - Interpleural analgesia in post-cholecystectomy patients--a clinical study. PMID- 8413066 TI - Mental health services in the context of health insurance reform. AB - Health care reform offers opportunities to improve the care of persons with serious mental illness, but it also can lead to the disruption of innovative systems of care that have been developed in recent years through Medicaid and other public programs. The care of mentally ill persons must be organized to maximize possible trade-offs between inpatient and other community services in a way that will promote function and satisfactory adjustment. An indemnity approach emphasizes controls on demand; instead, better design of supply-side approaches is required, using incentives to integrate services through case management and other methods. Considerable capacity development is needed to integrate long-term care appropriately into the mainstream, but serious barriers include control of risk selection and the difficulties of fairly adjusting capitation rates for high utilization patients. Issues in mental health care are paradigmatic of many other areas of medical care and reflect changing family and community organizations and new challenges in care and rehabilitation. PMID- 8413067 TI - Economic incentives in the choice between vaginal delivery and cesarean section. AB - The dramatic rise in cesarean-section (C-section) rates, and their high costs and wide variation, has raised interest in understanding the factors affecting decisions to use this procedure. The economic incentives of physicians, hospitals, payers, and mothers are examined. In the economic framework, physicians must balance their short-term interests against their reputation, which is derived from efficiently providing what mothers want. Providers who encounter higher opportunity costs while attending to mothers in prolonged labor can reduce these costs by operating or by restructuring their practices. The mainly indirect evidence on financial incentives indicates that insured mothers have low marginal financial costs when they undergo C-section. Mothers with private, fee-for-service insurance have higher C-section rates than mothers who are covered by staff-model HMOs, who are uninsured, or who are publicly insured. In conclusion, research and payment reforms to reduce distortions to good practice are proposed. PMID- 8413068 TI - From evidence to practice in the United States, the United Kingdom, and Canada. PMID- 8413069 TI - Preparing and updating systematic reviews of randomized controlled trials of health care. AB - People providing health care tend to turn to review articles rather than reports of primary research when seeking information to guide their practices. For this reason, the process of reviewing the results of primary research must respect scientific principles. Reviews must be kept up to date and disseminated in appropriate ways to the people who make decisions about health care, including policy makers, practitioners, and users of the health services. This article describes the methods developed and used in an attempt to address these challenges for care during pregnancy and childbirth. PMID- 8413070 TI - Retailing research: increasing the role of evidence in clinical services for childbirth. AB - A current review of the structures and assumptions of research transfer for clinical care reveals some progress from "passive diffusion" to "active dissemination" models, but little or no progress has been made toward targeting local influences on practitioner behavior for "coordinated implementation" of clinically relevant research into childbirth (or other) medical practices. The implementation of scientifically valid research syntheses, such as Effective Care in Pregnancy and Childbirth (ECPC), is therefore constrained by a poorly developed marketplace for retailing research information to practitioners. A survey in Canada of the four most significant potential retailing groups demonstrated that whereas clinical and community groups were adopting the necessary knowledge and attitudes, public policy makers and administrators trailed well behind them. To increase the probability of thorough retailing of ECPC, a three-phase plan could be instituted that would identify product champions within potential retailing groups, develop implementation activities for each retailing group, and convene annual conferences. PMID- 8413071 TI - Improving the use of research-based evidence in policy making: effective care in pregnancy and childbirth in the United States. AB - The rigorous syntheses in Effective Care in Pregnancy and Childbirth (ECPC) raise the question of how these and future recommendations can effectively be used to influence the decisions of policy makers and to improve the care we receive. The fragmentation of U.S. health care is especially striking for pregnancy and childbirth care. Although the U.S. context does not contain an obvious focal point for ECPC, several organizations have leverage or influence over different facets of policy making in this area: professional associations, consumer advocates, regulatory bodies, and third-party payers. An alternative to current independent activities is the organization of a network to facilitate implementation of ECPC. The intention would be that, through interaction, members would strengthen their interest and improve their effectiveness in implementing ECPC. An initial activity might be to evaluate alternative methods for implementing a specific recommendation. PMID- 8413072 TI - Implementing the findings of Effective Care in Pregnancy and Childbirth in the United Kingdom. AB - Effective Care in Pregnancy and Childbirth (ECPC) is a comprehensive analysis of evidence. The findings are only now beginning to influence practice in the United Kingdom several years after its publication. This article assesses the extent to which policy makers and practitioners are prepared to use evidence and proposes ways to implement the ECPC findings. Such efforts would require leadership from the U.K. Department of Health and the commitment of the leaders of professional and consumer bodies to promote perhaps 10 or 12 key findings rather than try to push forward the whole program at once. A coordinated approach would then be needed to influence managers, professionals, and service users, first through informing them of the findings, and then by devising specific approaches, such as the use of opinion leaders, for each group. Finally, changes in practice should be monitored. PMID- 8413073 TI - The prospect of data-based medicine in the light of ECPC. AB - A new effort in medical information has gathered and organized data from randomized clinical trials and other sources in obstetrics and made them available in several forms. This accomplishment implies the possibility of data based medical practice that depends on systematic organization of medical findings. Possible limitations in data-based medical practice are treated and the benefits of such an approach are discussed. Will it be possible and useful to generalize this obstetric effort to all of medicine, and, if so, how? With the establishment of the Cochrane Centre, the United Kingdom has launched a program directed toward this end. New ideas for disseminating the gathered information are needed. Funding for core support will be needed both to create the medicine wide information and to maintain it. PMID- 8413074 TI - Behavioral and neuroendocrine effects of diethyl ether exposure in the mouse. AB - Diethyl ether has diverse behavioral effects and is known for its ability to stimulate stress hormones, yet little is known of the concentrations in which these effects occur. To more fully characterize these effects, adult male NIH mice were exposed to a range of concentrations of ether (1000-30000 ppm) in an inhalation chamber and both behavioral and neuroendocrine responses were assessed. When responding was maintained under FI-60 s schedules of milk presentation, 30-min exposures to 1000 ppm ether resulted in minimal behavioral effects, 3000-10000 ppm increased rates of responding over two-fold and higher concentrations decreased responding almost completely. Five-min exposures to the same range of concentrations resulted in concentration-related effects which were smaller than those produced by 30-min exposures. Exposure to a similar range of concentrations in naive mice increased adrenocorticotrophic hormone (ACTH) and corticosterone levels in a time- and concentration-dependent manner. Five-min exposures to 10000 ppm ether increased levels of ACTH from a baseline of 25.95 pg/ml to 310.5 pg/ml but did not affect corticosterone. Thirty-min exposures to the full range of concentrations of ether, increased corticosterone from control levels of 70 ng/ml to 418 ng/ml at 30000 ppm, and increased ACTH from control levels of 19.13 pg/ml to 80.5 pg/ml at 30000 ppm, in a concentration-dependent manner. The increase in ACTH for 30-min exposures was not as large as that observed for 5-min exposures at 10000 ppm, nor was it as large as that seen for corticosterone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413075 TI - Neurobehavioral toxicity of methanol reflected by operant running. AB - Eleven Long-Evans male rats were trained to respond for food delivery by running in wheels under a Fixed Ratio = FR 20 schedule of reinforcement. Each 360 degrees rotation counted as a single response. Three food pellets were delivered for each reinforcement. The wheels, which provided transverse rods to be gripped by the rats, were specifically designed to reflect motor deficits produced by neurotoxicants. Each animal received two replicates of three different doses of methanol (50% in water): 1.0, 2.0, and 3.0 g/kg by gavage. The sequence for each animal was determined by a counterbalanced design. Gavage was followed by admitting the animal to the running wheel compartment 10 min later. Running wheel sessions lasted for 1 hour daily and were conducted 6 days/week. Statistical analyses showed insignificant differences between water and no-treatment control days, indicating no effect of the gavage procedure. However, a dose-effect relationship between methanol dose and responses per session proved statistically significant and linear (p < 0.0001) down to a dose equivalent to 10% of the LD50. In addition, detailed analyses of intervals between successive rotations (IRTs) indicated a displacement of the distribution toward longer intervals (decreased velocities) with increasing dose. The absence of a corresponding rise in the incidence of long pauses suggested that impaired coordination, reduced endurance, or their combination, rather than nonspecific variables, accounted for these results. PMID- 8413076 TI - Lack of effect of chronic developmental lead treatment on biogenic amines and metabolites in monkey cerebrospinal fluid. AB - Concentrations of 3,4 dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), norepinephrine (NE), 3-methoxy-4-hydroxyphenylglycol (MHPG), and 5 hydroxyindoleacetic acid (5-HIAA) were assayed in the cerebrospinal fluid (CSF) of control and chronically lead-treated nursery-reared rhesus monkeys sampled periodically from infancy through adulthood. Blood lead levels peaked at 62 micrograms/dl at 1.5 months of age, averaged 45 micrograms/dl for the remainder of the first year postpartum, and were maintained at 14 micrograms/dl from 20-58 months of age. Cisternal CSF samples were collected monthly from 5-35 months of age and every 1-4 months from 36-58 months of age. Biogenic amine and metabolite concentrations were assayed by high-performance liquid chromatography with electrochemical detection. Overall concentrations of DOPAC, HVA, NE, MHPG, and 5 HIAA were not significantly different in the control and lead-treated groups nor were there any significant interactions between lead treatment and age for any measure. DOPAC, HVA, and 5-HIAA concentrations decreased gradually with age, whereas MHPG concentration decreased sharply between 35 and 40 months of age. NE concentration remained stable across development. PMID- 8413077 TI - Bitertanol, a triazole fungicide, increases operant responding but not motor activity. AB - Several recent reports indicate that triadimefon, a triazole fungicide, has effects on behavior that are similar to those of psychomotor stimulants. For example, triadimefon increases overall fixed-interval (FI) response rate, disrupts FI response patterning, increases motor activity, and produces stereotypies at high doses. The present study was designed to determine whether similar behavioral effects on FI performance and motor activity could be produced by another triazole fungicide, bitertanol. The effects of bitertanol (10-300 mg/kg, IP) were determined in rats on performance maintained under a multiple FI 1-min FI 5-min schedule of reinforcement. Intermediate doses of bitertanol increased response rates and disrupted response patterning in both FI components. A second experiment determined the effects of the same doses of bitertanol on motor activity. In contrast to its effects on operant responding, bitertanol did not increase motor activity at any of the doses tested. These findings indicate that the behavioral similarities between bitertanol and triadimefon are limited and that a dissociation exists between biteranol's effects on operant performance and motor activity. PMID- 8413078 TI - Delay in the development of amygdala kindling following treatment with 3,3' iminodipropionitrile. AB - A disruption in cognitive function and induction of astrogliosis in the brain have been reported as neurotoxic effects of 3-3'-iminodipropionitrile (IDPN). Kindling is a model of synaptic plasticity that produces functional and structural alterations in brain that may encompass those underlying learning and memory. We examined kindling development in rats exposed to IDPN as neonates (0 or 225 mg/kg on postnatal days 5-7) or as adults (0 or 200 mg/kg/day for 3 days at 2-3 months of age). As adults, animals were electrically kindled in the amygdala with once daily stimulation until fully generalized seizures were evoked. IDPN significantly retarded the rate of development of kindling and shortened the mean afterdischarge (AD) duration with successive stimulations in animals dosed as adults. Neonatally-treated animals were not significantly slower to kindle but did demonstrate protracted AD development. No effect on pentylenetetrazol-induced seizures was seen. IDPN's reported effects on biogenic amines or its olfactory toxicity cannot readily account for the delay in kindling. Alternatively, we propose that disruption of synaptic transmission consequent to IDPN-induced neuronal degeneration in the CNS may contribute to an impairment of kindling. PMID- 8413079 TI - Prenatal nicotine exposure and cognitive performance in rats. AB - In humans and animal models there is evidence that prenatal nicotine exposure causes lasting deficits in cognitive performance. The current study examined the cognitive effects of prenatal exposure of rats to 2 mg/kg/day of nicotine. This dose did not cause significant deficits in maternal weight gain, offspring litter size, or pup weight. The control offspring showed the normal ontogeny of spontaneous alternation from near chance (50%) performance to 80%-85% alternation. In contrast, the nicotine-exposed rats had the opposite progression with abnormally high alternation on days 22-30 and abnormally low alternation on days 35-52. Acquisition of choice accuracy performance on the radial-arm maze (RAM) was not altered in a major way by nicotine exposure. Minor nicotine-induced changes in choice accuracy were seen during the initial trials of acquisition. The nicotine exposed female offspring had a significantly longer response duration. Prenatal nicotine exposure did significantly alter the effects of subsequent drug challenges on choice accuracy performance. The nicotine-exposed male offspring were significantly more responsive to the amnestic effects of the nicotinic antagonist mecamylamine. In a subsequent challenge, the effects of the beta-adrenergic antagonist propranolol were examined. A significant dose-related impairment in choice accuracy was seen in the control rats. In contrast, the nicotine-exposed rats did not show any significant response to propranolol. This decreased responsiveness to adrenergic challenge parallels the reduction in adrenergic response to nicotine challenge we previously found in littermates to the rats of the current study. Prenatal nicotine exposure causes subtle alterations in cognitive performance that can be magnified by challenges of nicotinic and adrenergic systems. PMID- 8413080 TI - Environmental enrichment and the behavioral effects of prenatal exposure to alcohol in rats. AB - Animals exposed prenatally to alcohol (4 g/kg/day) via maternal peroral intubation or control offspring were reared after weaning either alone in standard steel/wire cages or in groups of eight, for 6 weeks. Rats exposed prenatally to alcohol and reared in isolation had a dysmetric stride length indicative of an ataxic gait. However, following postweaning environmental enrichment, prenatal alcohol-exposed rats showed no evidence of ataxia. In addition, the prenatal alcohol-exposed rats showed the same magnitude of improved Morris maze performance after enrichment as did the control offspring. These preliminary results suggest that postnatal environment can influence the expression of alcohol-related birth defects in rats, that rats exposed prenatally to alcohol can benefit from the effects of enriched postweaning environment and that postnatal factors can attenuate some of the deficits due to prenatal alcohol exposure. PMID- 8413081 TI - Cocaine exposure during the brain growth spurt: studies of neonatal survival, somatic growth, and brain development. AB - Neonatal Sprague-Dawley rat pups were assigned to one of five groups. Three cocaine-treated groups were injected SC with either 40, 60, or 80 mg/kg/day of cocaine from postnatal day (PND) 4 through 9. Control groups were either injected with equivalent volumes of sterile dH2O (vehicle control) or received no injections (normal control) from PND 4 through 9. This early postnatal period, corresponding to the third trimester of pregnancy in humans, is characterized as a period of rapid development within the central nervous system (CNS), generally termed the brain growth spurt. The survival rate, somatic growth, and brain development in response to the various dosages of postnatal cocaine administration were assessed. There was a dose-dependent relationship between cocaine administration and survival rate. Furthermore, significantly reduced somatic growth, assessed in terms of body weight, was found in animals given 80 mg/kg cocaine daily, as compared with controls. With respect to brain weight, no significant differences were obtained among the various doses of cocaine-treated and control animals and there was no evidence of regional vulnerability (forebrain, cerebellum, or brainstem) to the cocaine insult. Additionally, neither an effect of gender, nor the interactions of gender with various doses of cocaine treatment was found on somatic growth and brain development. Taken together, the present results suggest that the brain exhibits a greater resistance to the cocaine insults than does somatic growth. Several possible explanations regarding the somatic growth retardation are discussed. PMID- 8413082 TI - Naltrexone does not increase brain and body development in rats. AB - Two strains of preweanling rats (88 Long-Evans; 120 Sprague-Dawley) were given daily SC injections of either 50 mg/kg naltrexone or the equivalent volume of saline, from birth to postnatal day 21. Brain and body weights were assessed at the end of the injections, and in Long-Evans animals, significant differences were noted on both measures with the outcome favoring the control animals. No significant differences were observed between the naltrexone and control Sprague Dawley rats. These results disconfirm the hypothesis that increased postnatal growth is accomplished by prolonged early postnatal opioid receptor blockade. PMID- 8413083 TI - C. Randall Nelms, M.D. The new MMA president is all ears. PMID- 8413084 TI - Informed consent. PMID- 8413085 TI - The information solution. Public, private efforts seek to improve care, reduce costs through data collection. PMID- 8413086 TI - Results of the Committee on Women Physicians' survey. PMID- 8413087 TI - Asthma in childhood. A review. AB - Asthma is currently considered by many to be a disease out of control. The prevalence and severity of asthma have increased in recent years, especially in children. Asthma is a very diagnosable condition. Better methods for treatment and monitoring now exist. It is hoped that with the implementation of expanded knowledge of asthma, this very common condition in childhood can be brought under better control. PMID- 8413088 TI - Data initiatives under MinnesotaCare. PMID- 8413089 TI - Injury in the medical workplace. Educating employees to prevent back injury, stress, and slips and falls. PMID- 8413090 TI - Health care reform. Physicians and medical societies must lead the way. PMID- 8413091 TI - The numbers game. Quantifying quality of care. PMID- 8413092 TI - Goodbye to the old-fashioned family doctor. PMID- 8413093 TI - The 'unacceptable circumstances' confronting physicians. PMID- 8413094 TI - [The Italian Society of Dentistry and Maxillofacial Surgery (SIOCMF), Naples, 15 June 1993]. PMID- 8413095 TI - [Malignant mesenchymal neoplasms of the oromaxillofacial area. The diagnostic problems]. AB - Oral and maxillo-facial mesenchymal malignant neoplasms are rare. They involve nosologic, diagnostic and therapeutic problems. Clinical cases of soft and bone tissue are reported. They share difficulties in clinical, roentgenographic and histopathological diagnosis and often the disagreement between the clinical evidence and the histopathological report. The rule of the immunohistochemistry for a correct characterization of mesenchymal malignant neoplasms and the necessity for correct and early diagnostic definition in order to plain the treatment are emphasized. PMID- 8413096 TI - [The classification and histopathological aspects of jaw cysts: an update]. PMID- 8413097 TI - [A clinical analysis of a group of Down's patients]. AB - The authors report their experience of the clinical analysis of 40 handicapped subjects: 28 suffered from various degrees of mental handicap and 12 had Down's syndrome. In particular, the authors focused their attention on anomalies of the stomatognathic apparatus in Down subjects, and examined and compared all those factors which are normally attributed to this pathology. An analysis of these findings shows a heterogeneity of clinical conditions which is not revealed in the literature. In the light of these new experiences, before applying pre programmed protocols in Down's patients it is important to carry out a careful control of their dental, muscular and functional situation together with the prospects for growth and development. PMID- 8413098 TI - [Multiple peg-shaped teeth associated with acrocephalosyndactyly. A variant of the Saethre-Chotzen syndrome? A clinical case]. AB - INTRODUCTION: The SC syndrome is relatively common among craniosynostosis syndromes, and it is transmitted as an autosomal dominant trait. The syndrome was first recognized and described by S. and C. (1931-1932), but that most extensive discussion of the disorder was published by Pantke et al. in 1975. He systematically described the most and the least common marks of that syndrome. CLINICAL CASE: We thought it right to report a case we met and observed, because together with the S.C. syndrome's marks, there was the presence of multiple peg shaped teeth. This peculiar mark is common to many orger craniosynostotic syndromes, but, we think it has never been described in "acrocephalosyndactyly type three" clinical cases. As a matter of fact, in literature, they have described in that syndrome, only lateral incisors shape anomalies. CONCLUSIONS: Even if in our patient the syndrome had a poor expressivity (there were scarce cranio-facial anomalies), and even if it came out there was no familiarity, it was peculiar because of the presence of multiple peg-shaped teeth. Besides, we think it is important to recognize that syndrome (which diagnosis is today still clinical because of the absence of peculiar laboratory aids), in such a way as to advice the female patient and to suggest her, in case of pregnancy, to go to a Uman Genetic Service for a an advice. PMID- 8413099 TI - [Neurilemmoma (schwannoma) of the oral cavity. A report of 2 clinical cases]. AB - Neurilemmoma (schwannoma) is a benign, encapsulated perineural tumor of neuroectodermal derivation that originates from the Schwann cells of the neural sheath of motor and sensitive peripheral nerves; the etiology is still unknown. The tumor is normally solitary, smooth-surfaced, slow growing and generally asymptomatic. It may develop at any age and there is no gender predilection. Head and neck are one of the most frequent localizations, but intraoral development is quite uncommon. In this area the mobile portion of the tongue, and in a decreasing order, the palate, the cheek mucosa, the lip and gingiva are the most frequent locations. Although it origins from the nervous tissue, only in 50% of the cases a direct relation with a nerve is demonstrated. The term schwannoma has been attributed in the past either for neurofibroma and neurilemmoma. Their histogenesis remain anyway a controversial argument. Some authors think that both tumors originate from Schwann cells and perineural connective cells. Others think that the first one originate from perineural cells, while the latter from Schwann cells. Diagnosis is confirmed by microscopic examination. Neurilemmoma shows two different components: Antoni type A and Antoni type B tissue. The first one consists of Schwann cells arranged in compact, twisted bundles, associated with delicate reticulin fibres and spindle-shaped nuclei aligned in parallel rows forming a typical palisading pattern. Between the rows there are fine cytoplasmatic fibrils with acellular, eosinophilic masses called Verocay bodies. The second one is formed by irregularly arranged masses of elongated cells and fibers similar in appearance to neurofibroma, with areas of cystic degeneration and edema.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413101 TI - [The Italian Society of Dentistry and Maxillofacial Surgery (SIOCMF), Naples, 12 July 1993]. PMID- 8413100 TI - [A mucoepidermoid tumor of the mandible. A report of a clinical case]. PMID- 8413102 TI - [The evaluation of the efficacy of auxiliary retention in the preparation of partial crowns]. AB - An extracted maxillary human premolar was prepared for 3/4 artificial crown. Ten models of the tooth were made and different combinations of auxiliary retentive components were prepared on each model: proximal and lingual grooves, pins and boxes. For each model an artificial crown was cast cemented to it, mechanically tested for retention and graphed. Pins proved more retentive than grooves, being the latter more retentive when placed asymmetrically more than in symmetrical positioning. PMID- 8413103 TI - [An evaluation of the role of anxiety and vasoconstrictors in dental practice]. PMID- 8413104 TI - [An ultrastructural analysis of the peritubular dentin and of the tubular lumen in healthy teeth]. AB - In this study the authors examined some specimens of fractured dentine of embedded healthy teeth, of people aged between 20 and 30. Connecting the scanning electron microscopy to a system of computerised analysis of spectrometry (EDS) allowed the analysis of the qualitative composition of the specimens. Therefore, the authors described the tubular structure of the sound, the peri- and the intertubular dentine, the winding course of the dentinal tubules and in some specimens, the presence of cylindrical structures inside the tubules. The EDS analysis demonstrated that these tubular structures have the same composition as the intertubular dentine. Former studies described hollow cylindrical structures having a similar morphology solely as a response of the odontoblasts to white spots. On the contrary, in this study it was demonstrated that it is possible to find cylindrical structures which are definitely mineralized in sound teeth without caries. However, the presence of such formations is not concomitant with the disappearance of peritubular dentine which is a typical structural modification of the pathological advancement of the carious lesion. PMID- 8413105 TI - [Preformed round-section NiTi arch wires: the mechanical and clinical aspects]. AB - The authors emphasize the need to set up several products proposed by the manufacturers. Regarding to orthodontic wires, a suitable evaluation of their mechanical properties to fit them for clinical use is needed. After a short outline of the mechanical properties of the wires on the market, the authors focus their attention on the NiTi Japanese alloy superelastic wires. Therefore, an evaluation of the failure aspect of NiTi preformed arch wires with round section of 0.018" of diameter is reported. The tested wires underwent to clinical use as first arch wires, and were evaluated after 8 weeks from the beginning of the therapy, when they have shown failure at the control in the 8th week. After having cleansed and prepared the specimens, their failure zones have been observed by the SEM. The failure aspect was evaluated to check the reasons that produced it, basing on its surface characteristics. The aspect of the surface of the failure and the absence of permanent deformations showed that it is a fragile failure. The fragile failure occurs when materials are not able to tolerate plastic deformations and, typically concerning NiTi alloys constituent the tested wires, it joins the superelastic behaviour and the high wire resilience. It means that this wire is able to produce constant, continuous and light forces for a given stress range, and, furthermore, it is able to absorb energy without permanent deformations. Since 1971 NiTi wires have been on the market but without superelastic and crystallographic shape memory properties possessed by the wires produced by Furukawa El. Co. in 1978.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413106 TI - [An update of the classification and of the diagnostic criteria of oral lesions in HIV infection. The European Economic Community (EEC) and the Collaborative Center of the World Health Organization for the Oral Manifestations of HIV Infection]. PMID- 8413107 TI - [The incidence of craniomandibular disorders in patients with cervical dysfunctions. A clinico-statistical assessment]. AB - The aim of this research was to measure the incidence of craniomandibular disorders in a group of patients with functional-type cervical alterations. The group consisted of 50 patients undergoing treatment for disorders of the cervical sectors of the spine. Each patient was subjected to a medical examination to investigate the presence of CMD signs or symptoms. From the data statistical analysis a higher percentage of cases with muscular and joint pain, limited mouth opening, deviation and deflection, were found in comparison with the percentage found among the general population. This demonstrates an overloading of the entire masticatory apparatus. Joint noise was less frequent, probably due to its exclusion from our sample of patients with arthrosis-type degenerative pathology. PMID- 8413108 TI - [Analgesic efficacy and the tolerance for piroxicam-beta-cyclodextrin compared to piroxicam, paracetamol and placebo in the treatment of postextraction dental pain]. AB - INTRODUCTION: Acute postoperative pain is a common experience in oral surgery practice. Non-steroidal anti-inflammatory drugs (NSAIDs) are quite effective against mild to moderate pain and they are generally better suited in ambulatory outpatients than narcotic analgesics. The analgesic activity of piroxicam, a well known NSAID has been documented in many pain states. Piroxicam can be administered once daily because of its long half-life, but its absorption in the gastrointestinal tract is slow as it is its onset of action. Piroxicam-beta cyclodextrin (PBCD) is a new formulation of piroxicam which is the product of supermolecular encapsulation of piroxicam with the cyclic oligosaccharide beta cyclodextrin. PBCD is absorbed much faster than standard piroxicam, and its action as an analgesic is consequently more rapid. The purpose of this study was to assess the efficacy and the rapidity of action of piroxicam-beta-cyclodextrin in comparison with standard piroxicam, paracetamol and placebo following surgical extraction of impacted third molars. MATERIALS AND METHODS: The study population was composed of 32 patients of both sexes and in good health. To be included into the study, patients must have had third molar removal resulting in acute post surgical pain of at least moderate intensity. The patients were then randomly assigned to one of four treatment groups. PBCD 20 mg tablets; piroxicam 20 mg capsules; paracetamol 500 mg tablets, or placebo. The study was conducted according to a double-blind, double-dummy design. Pain intensity and pain relief were recorded at 0.5, 1, 1.5, 2, 3, 4 hours after a single dose of the study drugs, by means of a Keele-type rating scale. Rescue analgesics were not allowed before one and a half hour after taking the study drugs. A global evaluation of study drugs was expressed by patients at the end of the observation period. RESULTS: Treatment groups were homogeneous for demographic characteristics of the patients and for pain intensity at the time of medication with study drugs. All patients who received placebo requested supplemental analgesics, while none of the patients treated with the active drugs needed rescue analgesics. PBCD and paracetamol were comparable for their analgesic effect, while the time lag before a significant reduction of pain intensity with piroxicam was longer. Piroxicam and PBCD were superior to paracetamol because they showed a substantial analgesic effect through the 4-hour study duration, while paracetamol did not induce a complete relief from pain. DISCUSSION: One of the most commonly utilized model for the evaluation of analgesics is the third molar extraction pain. Our study clearly differentiated between active drugs and placebo. Furthermore, while PBCD and paracetamol showed a rapid effect, piroxicam was slow in inducing pain relief. The analgesic and anti-inflammatory activity of PBCD and piroxicam brought about the resolution of pain and inflammation consequent to the dental extraction. Paracetamol, a pure analgesic, was not equally active and pain persisted, even if at a low grade, throughout the observation period; probably this was due to local inflammation and edema. The results of our study appear to confirm the pharmacokinetic data on PBCD, which showed that therapeutic blood levels are reached faster with PBCD than with the standard piroxicam formulation. This results should be confirmed in studies with an adequate number of patients. PMID- 8413109 TI - [Dynamic magnetic resonance of the temporomandibular joint. Observations and findings]. AB - Condylar movements can be executed only if the TMJ morphology can satisfy functional needs of the masticatory apparatus. Articular components have to be in good relation with dento-skeletal functional anatomy: if this does not occur, mandibular dynamics may establish some functional stresses on the condylar head and the articular eminence. These overloaded structures and particularly their functional surfaces react to this situation to be remodelling and deformation of the articular cavity. In our work we have used a new method to analyse condylar dynamics using MRI. The reconstruction of mandibular movements, using this instrumentation, permits to make some useful observations on the functional capability of TMJ to adjust itself to different maxillofacial morphologies and, consequently, how this joint can fall in a dysfunctional and pathological condition. PMID- 8413110 TI - Psychiatric nursing and the emergency setting. PMID- 8413111 TI - Symptom management: inpatient nursing care of persons with schizophrenia. PMID- 8413112 TI - Psychiatric nursing and the context of care. AB - Our ability to provide nursing care for persons with severe mental illness is contingent on awareness of contextual factors and our ability to adapt and influence them. Briefer inpatient stays and more partial hospitalization programs must lead to changes in our theoretical orientation and types of intervention. Awareness of the shift to community mental health and the threat of the remedicalization of psychiatry should encourage nurses to broaden their roles and add new skills. Case management and political and systems advocacy are particularly crucial responses to the needs of the severely mentally ill and their families. We must never lose sight of individual patients and their pain. The alleviation or prevention of this pain is our ultimate goal. PMID- 8413113 TI - Residential care operators: perspectives on mental illness and caregiving roles. PMID- 8413114 TI - Beyond blame and shame: families coping with long-term mental illness. AB - Within this "Decade of the Brain," an accusatory veil lingers about the family's role in the genesis of mental illness. Health professionals need a framework to deal with chronic mental illness and caregiving. PMID- 8413115 TI - Familial touch and perceptual reactivity of children with severe mental illness. PMID- 8413116 TI - Caregiving and Alzheimer's-type disorders. PMID- 8413117 TI - A consumer-oriented program review process for mental health care. PMID- 8413118 TI - A nursing perspective on mental health policy. AB - Major mental health reform is under way to better serve the needs of people with severe mental illness. Reform will happen faster if psychiatric nursing wakes up from its long sleep to focus on services, programs, and systems of care for this population and, in so doing, sets a proper place for the discipline at the health policy table. PMID- 8413119 TI - [Verbal fluency and EEG coherence in Alzheimer's disease]. AB - EEG coherence of 16 derivations and a verbal fluency test were evaluated on 25 ambulatory patients suffering from Alzheimer's disease (age: 74 +/- 5.7). The aim of the study was to analyze coherence rate variations from different cortical areas in relation to the performance in the test. Coherence rates of each derivation with the other ones were calculated for four frequency bands from 0.5 to 13.5 Hz. Arithmetic averages of these rates were then calculated to obtain scalp averaged coherence rates. The patients were submitted to a verbal fluency test and divided into two groups according to their test score as compared to normative data: impaired (n = 10) and not impaired (n = 15). Results showed that averaged coherence rates of theta and alpha 1 frequency bands were significantly depressed in the impaired group in comparison with those of the not impaired group. Two cortical areas were concerned with this difference, the left temporo occipital and frontal zones, in an independent way for the same subject. Moreover, we observed that the patients who where impaired in the verbal fluency test, also presented a significantly greater ancientness of the clinical symptoms as compared to the not impaired patients. PMID- 8413120 TI - [Motor and staged sensory conduction velocities of the ulnar nerve in normal subjects]. AB - Normative data of ulnar motor and orthodromic sensitive conduction velocities are performed at different levels on 68 (19 to 49 year old) adult population which included 35 men and 33 women. In fact, 102 ulnar nerves are tested, 51 nerves for female population and male population respectively. For motor conduction velocities, the ulnar nerve is stimulated at five levels: at the wrist level, below elbow, at the elbow, above elbow, and at the axillary point; the motor responses were picked up on the hypothenar muscles (surface electrodes). For orthodromic sensitive conduction velocities, the fifth finger is stimulated and the electrical activities are picked up by surface electrodes which are placed at the same points. The difference between the conduction velocities values measured on the below elbow/wrist segment and the above elbow/wrist segment proved to be satisfactory for the possible detection of a compression of the ulnar nerve at the elbow. PMID- 8413121 TI - [Topographical study of event-related desynchronization: value in the psychophysiologic approach to cognitive processes]. AB - The interest of the event-related desynchronization study has already been evidenced, for example in the study of the cerebral activation modifications consecutive to cognitive processes. We observed here the evolution of this index during a recognition task where the material to memorize varied. Ten young subjects had to learn and then recognize a list of words or forms. The desynchronization of the alpha rhythm consecutive to each stimulus was recorded at 14 localizations on the scalp. We observed that the time evolution of the event-related desynchronization was different in each task. In the visual task, the phenomenon began sooner and was longer lasting. As long as the cortical area were concerned, any dominance nor interhemispheric difference were observed. During the visuospatial recognition task, the event-related desynchronization was however more widespread. PMID- 8413122 TI - Taking advantage of opportunities for change in learning disabilities nursing. PMID- 8413123 TI - Facing the future: credibility in a changing world. AB - As many rapidly changing and often conflicting influences come to bear on nurse education, and as nurse education strengthens links with higher education or integrates with this sector, nurse teachers need to reflect on their future role in terms of credibility. Four dimensions of credibility are suggested: teaching, knowledge, clinical and academic credibility. In considering these dimensions and the nature of future curricula, proposals for the future role are presented. It is suggested that the nurse teacher of the future must not only be a competent teacher, but a competent teacher of theory and practice. This implies not only teaching, but knowledge and clinical credibility. Academic credibility will be required by those who purchase nurse education and who will expect it to be at higher education level. It is concluded that the achievement of credibility across these four dimensions requires a degree of specialisation and that those who manage education must institute frameworks and practices to facilitate nurse teachers in their future role. PMID- 8413124 TI - Preparing the advanced practitioner in learning disabilities nursing. AB - This paper aims to describe the practical, therapeutic and educational dimensions of professional preparation in the field of Learning Disabilities Nursing. Through an exploration of the primary, secondary and tertiary dimensions of care, the authors identify the range of skills and competencies which they consider necessary for the achievement of advanced practice in this specialist field. Arguments are proposed which demonstrate the essential and continuing role of the nurse through a consolidation of the three dimensions of care. In so doing, an educational foundation for advanced practice which enables the nurse to work effectively within present and future health and social care initiatives is proposed. PMID- 8413125 TI - Developing a rationale for research based practice: some considerations for nurse teachers. AB - There is considerable interest in nursing literature regarding the development of research-based practice. This paper considers the implications for nurse teachers advocating a research focus on the problems that arise in practice utilising practitioner and action-based methodologies. The rationale for practitioner action-based approaches is outlined with consideration of the major claims made for the approach in educational literature. The paper concludes with a warning about the limitations of the approach and a statement of its contemporary relevance to nurse education. PMID- 8413126 TI - Curriculum issues related to the newly developed nursing diploma courses. AB - This paper discusses the findings related to curriculum issues in the newly developed nursing diploma courses. The study, of which this aspect is a part, explored the emerging role of the nurse teacher within the new programmes in England. Data were collected by a Delphi Survey followed by in-depth interviews with a sample of the Delphi respondents. The findings from the interviews revealed that the six colleges involved had all adopted a process approach to the development of their new curricular. The educational philosophy and the curriculum approach in each college focused on the development of the individual and the adult theory of learning. A strategy for curriculum evaluation was found to be in the early stages of development for which no model or framework had been identified. PMID- 8413127 TI - Decision-making in clinical practice: how do expert nurses, midwives and health visitors make decisions? AB - Decision-making is an essential and integral aspect of clinical practice. Preparation for clinical decision-making is haphazard and unplanned, in part because the process of making clinical decisions is not fully understood. This is one study of how expert nurses, midwives and health visitors make clinical decisions. The project involved a literature review and a series of workshops with expert practitioners to uncover the decision-making process in clinical practice. The study found that decision-making is an essential attribute of the expert practitioner, must be based on sound knowledge, may involve risk-taking and can only flourish in a supportive environment. Most importantly, clinical decision-making must take place within the context of a philosophy of care. Without such a philosophy, decisions will be arbitrary, uninformed and probably unsafe. PMID- 8413128 TI - Building on strengths through preceptorships. AB - Learning needs of registered nurses in post-RN degree programs vary from those of basic (generic) baccalaureate or diploma nursing students. Despite this awareness, there is a paucity of literature which addresses the utility of preceptorships with this group. This paper discusses various viewpoints regarding the preceptorship process experienced by one post-RN degree nursing program in Western Canada. Specifically, the paper describes how the process was used with the post-RN population and discusses outcomes from student, faculty and preceptor perspectives. PMID- 8413129 TI - Change the curriculum--or transform the conditions of practice? AB - This paper explores the notion that contemporary teaching practices reinforce and maintain the legitimacy of traditional relations of power between teachers and students of nursing. Nurse teachers and clinicians have socially constructed and legitimated power over students which acts to constrain the development of critical consciousness. Student-centred learning packages and strategies such as problem-solving, questioning and dialogue may give the impression of student empowerment while leaving the authoritarian nature of teacher-student relationships intact. Furthermore nursing education is premised on the belief that 'real' learning takes place in the classroom (where teaching occurs) and is consolidated by practice (where nursing occurs). This situation creates a major dilemma for all teachers since the contradictions between classroom knowledge and experiential clinical knowledge are seldom officially recognised. The rhetoric of critical social science however, suggests that emancipation and empowerment of teachers and students would follow their enlightenment as to the nature of these contradictions. This assumption discounts the ways in which hegemonic ideology shapes the consciousness of nurses to accept dominant views of what constitutes professional practice or legitimate knowledge and how that may be obtained. PMID- 8413130 TI - A study of attitudes to gender and nursing stereotypes in newly recruited student nurses. AB - This article presents the findings of a study which investigated the attitudes of 100 newly recruited student nurses towards gender and nursing stereotypes. The students were in their second day of the Common Foundation Programme at North Staffordshire College of Nursing and Midwifery. The data were collected using a researcher developed instrument which utilised a Likert scale for measurement of attitudes in relationship to statements pertaining to gender and nursing stereotypes. The findings reveal overall high mean scores and an implied propensity towards beliefs in gender and nursing stereotypes. Analysis related to demographic variables of gender, age and sex gives a number of significant results. The highest proportion of significant results were related to gender differences in the sample. The data provide a basis for evaluation of teaching and learning related to students in the sample course of studies. The findings also have more general implications for curricular issues and the development of teaching strategies. PMID- 8413131 TI - Project 2000: the gap between theory and practice. AB - This article explores the theory-practice gap in the context of Project 2000. Drawing on the findings of empirical research into the implementation of Project 2000 in one district health authority, we conclude that tensions between education and service staff have far from disappeared with the advent of Project 2000. The article discusses firstly the difference in emphasis between education and service staff over the question of equipping students with practical skills during their Common Foundation Programme. Secondly, we discuss the fact that whilst educationalists are intent on fostering self-directed learning in students, service staff are more concerned with producing safe practitioners. The article concludes on an optimistic note, suggesting that although service and education staff may differ in the relative importance they attach to teaching students particular types of skill; and although they may be operating on different timescales, fundamental disagreements between the two groups are absent. PMID- 8413132 TI - Maintenance of current clinical knowledge is essential to nurse teachers--based on the situation in China. AB - During the last decades, especially since the 1970s, an abundance of nurse education programs have been established in The People's Republic of China. Nurses have better opportunities for education than ever before. It is common knowledge that the quality of the student nurse is closely related to the quality of the nursing teaching. The level of teaching depends on faculty members' experience as well as on instructional material. Furthermore, it is important that the teaching is related to practical situations. Based on necessities, however, the instructional material can be changed by the educators. Consequently, faculty members are essential to the improvement and enhancement of the teaching quality. This paper has been written with a view to the present situation of nursing education in China in order to emphasize the importance of maintaining and renewing clinical knowledge to nurse teachers. PMID- 8413133 TI - 'But we're doing it already!' Exploring a response to the concept of reflective practice in order to improve its facilitation. AB - The paper uses the stimulus of a personal reflection to explore how the concept of reflective practice can be facilitated. The thoughts on facilitation combine the use of recognised knowledge, experience and reflection to offer a way of reducing the risk of rejection of reflective practice. In setting the context for such an exploration, reflective practice is defined and its potential contribution to nursing considered. PMID- 8413134 TI - The development of a BA in community health nursing/BA in nursing by distance learning. AB - A new degree course in community health care nursing has been developed at The Robert Gordon University, with validation having taken place on 30 June 1992. It is innovative as it is being offered to students by a part-time distance learning route only, over a minimum time of 18 months and a maximum time of 3.5 years, and is replacing the full-time diploma courses offered in district nursing, health visiting and occupational health nursing. As well as the community health nursing route, a BA in nursing has also been developed for any registered nurse who wishes to top-up to degree level. PMID- 8413135 TI - Congenital cystic adenomatoid malformation. AB - Congenital cystic adenomatoid malformation (CCAM) is an infrequently seen abnormality. The pathology of the various types has been well described, but the pathogenesis of the defect is less clear. A review of the literature revealed a lack of nursing information relevant to the disease itself. This article presents a review of the pathophysiology of CCAM, medical management, and differential diagnosis in addition to the intricate care involved in nursing an infant with this defect. PMID- 8413136 TI - Congenital hypothyroidism: diagnosis and management. AB - Every newborn infant should be screened for congenital hypothyroidism before discharge from the nursery. Neonatal screening for hypothyroidism was introduced to North America in 1972 and has been demonstrated to be cost effective in the prevention of neurological damage in children. Interpretation of diagnostic test results from such programs is based on understanding the physiology of the thyroid gland and recognition that neonatal hypothyroidism primarily stems from an embryological disorder of thyroid development rather than central causes involving the hypothalamic-pituitary axis. Early diagnosis of hypothyroidism is dependent on proper timing and collection of blood samples and an efficient screening program reporting accurate results. A thorough maternal and family history in conjunction with clinical signs and symptoms of hypothyroidism, biochemical tests, and radiological findings should be used to rapidly establish the diagnosis. Early detection and treatment with thyroxine normalizes skeletal maturation, physical growth, cognitive functioning, and motor development of affected newborns. Nurses play an important role in identification, management, and supportive care of infants with hypothyroidism so that maximal potential is achieved. PMID- 8413137 TI - Deciding what to do when the patient can't speak: a preliminary analysis of an ethnographic study of professional nurses in the neonatal intensive care unit. AB - The role of patient advocate is not easily implemented with infants in the neonatal intensive care unit because there is no way of knowing what the infant would wish to be done. Many of the nurse's actions are based on the concept of the "best interests of the infant." However, although widely used, this concept is rarely defined or described for the neonatal population. The purpose of this qualitative study was to identify themes related to the professional nurse's interpretation of the concept of advocacy. A case study design and inductive data analysis consisting of constant comparison and triangulation methodologies were used. This research resulted in multiple perspectives of a single phenomenon based on the similarities and differences in the nurse's perspective related to the concept of interest. PMID- 8413138 TI - Skin-to-skin (kangaroo) holding of the intubated premature infant. AB - Skin-to-skin holding has been reported to be a useful technique for helping mothers feel close to their nonintubated infants hospitalized in the NICU. This article describes our experience with skin-to-skin holding of 25 intubated infants in the NICU. We found this technique was safe for the intubated infant and promoted parental attachment, even in parents who were at high risk for attachment impairments. Our experience suggests that skin-to-skin holding with small intubated infants may offer some parents an effective method to overcome some of the barriers to attachment imposed by the infant's hospitalization. In addition, our observations raise several questions regarding parent-infant interaction. PMID- 8413139 TI - Isn't it time to do the right thing? PMID- 8413140 TI - The development of a tool to assess neonatal pain. AB - The objectives of this study were to (1) develop a behavioral assessment tool for the measurement of pain in the preterm and full-term neonate; (2) establish the construct and concurrent validity, interrater reliability, and internal consistency of the tool; and (3) examine the relationship between the pain scores and infant characteristics. Thirty-eight infants contributed to the 90 procedures videotaped for the study. The Neonatal Infant Pain Scale (NIPS) was used to score behavioral responses before, during, and after each intrusive procedure. The significant difference in NIPS scores over time indicates that the scale provides a measurement of intensity of infant responses to intrusive procedures. Concurrent validity was established by correlations, ranging from .53 to .84, between NIPS scores at each minute of observation and scores on the Visual Analogue Scale. Interrater reliability was high: Pearson correlations ranged from .92 to .97 across successive minutes of observation. The six component scores of the NIPS had high internal consistency: Cronbach's alphas were .95, .87, and .88 for before, during, and after the procedures, respectively. Although gestational age and five-minute Apgars were positively associated with NIPS scores over time, there was no association between these factors and responsiveness to pain, as measured by change in NIPS scores from before to during the procedure. Results are discussed in terms of the use of the NIPS in clinical trials and its clinical application in a neonatal intensive care unit. PMID- 8413142 TI - Mechanics of ventilation: resistance. PMID- 8413141 TI - Maternal views of preterm infants' responsiveness to social interaction. AB - Mothers of 47 very low birth weight ( < 1,500 gram) infants were interviewed concerning their feelings and views about interactions with them. Interviews were conducted within the first ten days and again three to five weeks after birth. Over the first month of the baby's hospitalization, mothers significantly increased their pleasure in interacting, knowledge of infant cues, and perception that their infants were responding. The most frequently mentioned behaviors were maternal talking and touching and infant eye opening and body activity. Many mothers appeared to be utilizing their infants' behaviors as a guide for their own behaviors and reported that their infants' behaviors had specific meanings. However, an equal number of mothers were not ascribing meaning to their infants' behaviors and did not appear to be using behavioral cues. These findings have implications for interventions by neonatal nurses. PMID- 8413143 TI - Procedural flowchart job aid for chest tube placement. PMID- 8413144 TI - Sexual harassment in the workplace. PMID- 8413145 TI - Instruments in neonatal research: measuring pain in the preterm infant. PMID- 8413146 TI - A review of gentamicin use in neonates. AB - Gram-negative sepsis contributes significantly to neonatal morbidity and mortality. Gentamicin is an aminoglycoside antibiotic used in the treatment of gram-negative infections. However, developmental differences of the neonate (compared with the older child or adult) influence the drug's disposition in the body. Administration, distribution, elimination, as well as susceptibility to toxicities may be altered in the neonatal period because of these pharmacokinetic differences. A literature review reveals pharmacokinetic differences of the neonate that affect gentamicin dosing. Nursing considerations affected by the developmental differences of the neonate include knowing appropriate dosages and routes of administration, pathophysiological and pharmacological conditions that affect gentamicin disposition, serum monitoring, and evaluation of adverse reactions and toxicities. PMID- 8413147 TI - Neonatal glucose determinations obtained from an umbilical artery catheter: evaluation for accuracy using an in vitro model. AB - Accuracy of glucose values is crucial in the management of the critically ill neonate. Sampling for these values may be obtained from an umbilical artery catheter (UAC), but dextrose solutions infused through these catheters may contaminate samples obtained for glucose determination. An in vitro study was conducted to evaluate the effect of blood withdrawal amount and normal saline irrigation on the accuracy of glucose measurements obtained from a UAC. Methods included randomization of blood withdrawal amounts and irrigation with normal saline solution prior to obtaining the blood for glucose measurement. Two hundred and eighty glucose samples were obtained. Results revealed a significant difference between measured and baseline glucose values for all withdrawal amounts and varying flush volumes. This inability to approximate the baseline glucose values could mean that a UAC sample may not reflect the neonate's actual glucose value. PMID- 8413148 TI - Collaborative experiences for NICU and early childhood education personnel. AB - Participation of NICU staff in special education teacher training is discussed as a method for improving follow-up care of high-risk infants and for helping agencies comply with Public Law 99-457, which mandates services for disabled preschoolers and encourages intervention for infants and toddlers. Through a collaborative training program involving a medical center and a university, education students spent 15 to 20 hours in the NICU. Nurses gained a greater understanding of community-based early intervention programs and available providers and made referrals with more confidence. Student teachers witnessed the medical conditions and treatments experienced by NICU patients, gaining an understanding of the types of medical histories their future clients would have. The collaborating hospital developed working relationships with agencies that provide follow-up care to NICU graduates. The article discusses medical and educational models of care, implications and benefits of incorporating developmental care into NICU practice, effects of the law on early intervention programs, and features of the model collaborative program. PMID- 8413149 TI - Stabilization and transportation of the infant with PPHN. AB - Infants with persistent pulmonary hypertension of the newborn (PPHN) present special nursing challenges. This article reviews signs and symptoms of this disease process and explores the implications of care for the referring facility during transport and subsequent tertiary care. An overview of basic physiology, fetal circulation, and diagnostic testing is incorporated. Associated complications and future trends are addressed. PMID- 8413150 TI - Memo: from baby to my nurse. PMID- 8413152 TI - Methodological issues: the measurement of long-term outcome. PMID- 8413151 TI - Pulmonary function testing: a tool for managing the mechanically ventilated patient. PMID- 8413153 TI - Chlorothiazide. PMID- 8413154 TI - Premature infant diapers. PMID- 8413155 TI - Understanding neonatal chest x-rays. Part I: What to look for. AB - Understanding chest x-rays is an important aspect of nursing care. It enables the nurse to (1) recognize and report abnormal findings that may be life threatening, (2) follow the progression of a disease process, and (3) optimize patient care. A systematic approach is necessary when reading a chest x-ray to reduce the possibility of missing important features, especially in the presence of an obvious or striking abnormality. This article discusses several factors to consider when reading a chest x-ray. It also reviews the clinical and radiographic manifestations of some neonatal respiratory disorders. PMID- 8413156 TI - Hantavirus infection--southwestern United States: interim recommendations for risk reduction. Centers for Disease Control and Prevention. AB - This report provides interim recommendations for prevention and control of hantavirus infections associated with rodents in the southwestern United States. It is based on principles of rodent and infection control and contains specific recommendations for reducing rodent shelter and food sources in and around the home, recommendations for eliminating rodents inside the home and preventing them from entering the home, precautions for preventing hantavirus infection while rodent-contaminated areas are being cleaned up, prevention measures for persons who have occupational exposure to wild rodents, and precautions for campers and hikers. PMID- 8413157 TI - Initial therapy for tuberculosis in the era of multidrug resistance. Recommendations of the Advisory Council for the Elimination of Tuberculosis. AB - These recommendations update previous CDC/American Thoracic Society (ATS) recommendations for the treatment of tuberculosis (TB) among adults and children. The most notable changes are in response to the increasing prevalence of drug resistant TB in the United States. These recommendations include the need for a) in vitro drug susceptibility testing of Mycobacterium tuberculosis isolates from all patients and reporting of these results to the health department, b) initial four-drug regiments for the treatment of TB, and c) initial directly observed therapy for persons with TB. Adherence to these recommendations will help prevent the occurrence of more cases of drug-resistant TB, reduce the occurrence of treatment failure, and reduce the transmission of TB in the United States. PMID- 8413158 TI - Prevalence of work disability--United States, 1990. AB - Work disability, defined as the inability to perform work as a result of a physical, mental, or other health condition, costs approximately $111.6 billion each year in direct and indirect medical costs and lost wages. National health objectives for the year 2000 are to increase the span of healthy life for persons in the United States and to reduce the proportion of persons experiencing disability from chronic conditions (as defined by CDC's National Health Interview Survey) to a maximum of 8% (baseline: 9.4% in 1988) (objective 17.2). This report presents national and state-specific prevalence rates of work disability in the United States for 1990 and compares rates with those for 1980. PMID- 8413159 TI - Prevalence of mobility and self-care disability--United States, 1990. AB - An estimated 43 million persons in the United States have a disability; the estimated annual economic impact of disabilities--representing loss of wages, medical-care expenditures, and additional household expenditures--is approximately $176.7 billion. The Institute of Medicine recently recommended surveillance and systematic collection of information at the national and state levels to assist in program planning and evaluation for state-based programs for the prevention of disabilities and secondary conditions (i.e., health conditions resulting from a disability). To characterize state-specific disability patterns and better plan for funding of disability services, the National Institute of Disability and Rehabilitation Research (NIDRR) and CDC assessed data from the 1990 census on two forms of disability: difficulty with mobility and self-care activities. This report summarizes the results of the assessment for persons aged > or = 16 years. PMID- 8413160 TI - Comprehensive delivery of adult vaccination--Minnesota, 1986-1992. AB - Despite the availability of safe and effective vaccines, many adults still suffer from vaccine-preventable diseases. For example, each year an estimated 40,000 60,000 adults die as a result of pneumococcal infection and influenza. In addition, from 1985 through 1992, 433 (92.7%) of 467 cases of tetanus occurred among adults (CDC, unpublished data, 1993). Although up to 90% of influenza related deaths occur among persons aged > or = 65 years, the 1991 National Health Interview Survey indicated that, during the preceding year, only 41% and 20% of persons aged > or = 65 years reported receiving influenza vaccine and pneumococcal vaccine, respectively (CDC, unpublished data, 1991). This report describes the efforts of the Hennepin County (Minneapolis) Community Health Department (HCCHD) (1990 population: 1.1 million) to provide comprehensive vaccination services to persons aged > or = 62 years. PMID- 8413161 TI - Progress in the development of hantavirus diagnostic assays--United States. PMID- 8413162 TI - Violence-related attitudes and behaviors of high school students--New York City, 1992. PMID- 8413163 TI - Unintentional carbon monoxide poisoning from indoor use of pressure washers- Iowa, January 1992-January 1993. PMID- 8413164 TI - Self-reported HIV-antibody testing among persons with selected risk behaviors- southern Los Angeles County, 1991-1992. PMID- 8413165 TI - Outbreaks of Salmonella enteritidis gastroenteritis--California, 1993. AB - Foodborne infections cause an estimated 6.5 million cases of human illness and 9000 deaths annually in the United States. Salmonella is the most commonly reported cause of foodborne outbreaks, accounting for 28% of such outbreaks of known etiology and 45% of outbreak-associated cases during 1973-1987. During 1985 1992, state and territorial health departments reported 437 Salmonella enteritidis (SE) outbreaks (Table 1), which accounted for 15,162 cases of illness, 1734 hospitalizations, and 53 deaths. This report describes three SE outbreaks in California during a 4-month period in 1993. PMID- 8413166 TI - Morbidity surveillance following the midwest flood--Missouri, 1993. AB - Heavy spring and summer rains during 1993 caused flooding by both the Mississippi and Missouri rivers and by streams in 84 of the 115 counties in Missouri; all 84 were declared federal disaster areas. The Mississippi River attained flood stage during July 15-August 2, and the Missouri River, during July 25-August 2; a total of 2,060,757 acres were submerged, and approximately 60,000 persons throughout Missouri were displaced (National Weather Service, unpublished data, 1993). At the request of the Missouri Department of Health, CDC provided assistance in implementing a surveillance system to monitor flood-related injuries and illnesses in the affected areas. This report presents preliminary findings of these surveillance efforts. PMID- 8413167 TI - Human rabies--New York, 1993. AB - In August 1993, a fatal case of human rabies in an 11-year-old girl was reported to the New York State Department of Health; this was the first indigenously acquired fatal case diagnosed in New York in 39 years. This report summarizes the investigation of this case. PMID- 8413168 TI - Pregnancy outcomes following systemic prenatal acyclovir exposure--June 1, 1984 June 30, 1993. AB - Herpes infections are common among women of reproductive age (i.e., aged 15-44 years). Acyclovir (Zovirax), an antiviral drug effective in the treatment of herpes simplex infection, was approved by the Food and Drug Administration (FDA) in 1984. Since its approval, the effects of acyclovir on human pregnancies have not been determined. However, inadvertent pregnancy exposures to acyclovir were expected to occur among women in whom treatment had been indicated for preexisting herpes simplex infections. Some physicians have reported intentional use of acyclovir during pregnancy for treatment of life-threatening herpes simplex infection. To assess the outcomes of pregnancies exposed to acyclovir, the Acyclovir in Pregnancy Registry was established on June 1, 1984, by the manufacturer, in collaboration with CDC. This report summarizes data on pregnancies reported to the registry through June 30, 1993. PMID- 8413169 TI - Measles--United States, first 26 weeks, 1993. AB - As of July 3, 1993 (week 26), local and state health departments had reported a provisional total of 167 measles cases for 1993--the lowest total reported for the first 26 weeks of any year since surveillance began in 1943 and a 99% decrease from the 13,787 cases reported during the first 26 weeks of 1990, the peak of the recent resurgence. Cases were reported from 18 states. This report summarizes the epidemiologic characteristics of measles cases reported for the first 26 weeks of 1993 and compares them with cases reported during 1989-1991. PMID- 8413170 TI - Update: hantavirus pulmonary syndrome--United States, 1993. AB - A unique hantavirus has been identified as the cause of the outbreak of respiratory illness (hantavirus pulmonary syndrome [HPS]) first recognized in the southwestern United States in May 1993. The habitat of the principal rodent reservoir for this virus, Peromyscus maniculatus (deer mouse), extends throughout most of the United States except the Southeast. Through October 21, 1993, HPS has been confirmed in 42 persons reported to CDC from 12 states (Figure 1). This report summarizes major clinical, pathologic, and diagnostic findings in patients with this newly recognized syndrome; addresses the use of the investigational antiviral drug ribavirin; and presents revised screening criteria for national surveillance. PMID- 8413171 TI - Home-health and hospice care--United States, 1992. AB - An estimated 9.5 million persons in the United States have difficulty performing basic life activities because of mental or physical health conditions. In recent years, an increasing range of home-care services--including home-health care and hospice care--have been created for persons requiring long-term care, and access to such care has been increased through public programs such as Medicare and Medicaid. To better characterize the use of these services in the United States, CDC's National Center for Health Statistics conducted the 1992 National Home and Hospice Care Survey (NHHCS), the first survey of home-health agencies and hospices and their patients. This report presents preliminary findings from the survey. PMID- 8413172 TI - Update: changes in notifiable disease surveillance data--United States, 1992 1993. AB - Since April 1990, CDC has graphically presented changes in reported cases of 14 notifiable infectious diseases reported through the National Notifiable Diseases Surveillance System (NNDSS). Figure I of each issue of MMWR displays on a log scale the ratio of the number of cases reported in the most recently ended 4-week period to that of the mean of the number of cases reported in 15 historical 4 week periods. During 1992-1993, Figure I has indicated a decline in incidence in selected reportable diseases (e.g., measles, mumps, and rubella). Declines in the incidence of these diseases could represent either true declines or changes in notifiable disease surveillance (e.g., a lower percentage of all cases was reported than previously). During June-August, 1993, CDC evaluated the surveillance system to determine the reasons for the declines. This report summarizes the results of the evaluation. PMID- 8413173 TI - National poliomyelitis immunization days--People's Republic of China, 1993. AB - In 1988, the World Health Organization (WHO) established the goal of global eradication of poliomyelitis by the year 2000. Based on cases officially reported to WHO, progress toward eradication has been substantial: in 1992, a total of 15,445 paralytic poliomyelitis cases were reported worldwide, compared with 32,419 cases in 1988. Beginning in December 1993, the People's Republic of China will conduct a series of two National Immunization Days that target approximately 100 million children (all children aged < 4 years) to receive oral poliovirus vaccine (OPV) in each of two separate rounds of vaccination, possibly representing the largest public health event of its kind in history. This report summarizes the plans for National Immunization Days and efforts in China to eradicate poliomyelitis by 1995. PMID- 8413174 TI - Diphtheria outbreak--Russian Federation, 1990-1993. AB - Despite high levels of vaccination coverage against diphtheria, an ongoing outbreak of diphtheria has affected parts of the Russian Federation since 1990; as of August 31, 1993, 12,865 cases had been reported. This report summarizes epidemiologic information about this outbreak for January 1990-August 1993, and is based on reports from public health officials in the Russian Federation. PMID- 8413175 TI - Pregnancy complications and perinatal outcomes among women with diabetes--North Carolina, 1989-1990. AB - Women with diabetes have a higher risk for complications of pregnancy than do women without diabetes; in addition, infants born to women with diabetes are at increased risk for adverse birth outcomes. Preconception counseling for women with established diabetes and early and continual prenatal care for women with established or gestational diabetes can reduce maternal and infant morbidity and mortality. Although the rate of pregnancy complicated by diabetes and the use of prenatal care varies by race of the mother, it is unknown whether the effect of diabetes on maternal and infant outcomes differs by race. Race reflects differing distributions of several risk factors for pregnancy outcomes (e.g., socioeconomic status and access to comprehensive health care) and is useful for identifying groups at greatest risk for adverse outcomes. To determine the prevalence of diabetes during pregnancy among women residing in North Carolina and to characterize differences in prenatal care and the risk for maternal complications and adverse pregnancy outcomes by race among mothers with diabetes, the North Carolina State Center for Health and Environmental Statistics examined birth certificates of infants of women who gave birth in the state during 1989-1990. This report summarizes the findings of the study. PMID- 8413176 TI - Behavioral Risk Factor Surveillance, 1991: monitoring progress toward the nation's year 2000 health objectives. AB - PROBLEM/CONDITION: Risk reduction is a major focus of the national health objectives for the year 2000. Progress toward several of these objectives can be evaluated by using data from the Behavioral Risk Factor Surveillance System (BRFSS). Year 2000 objective areas measurable by BRFSS data include those for overweight, lack of physical activity, smoking, safety belt use, and medical screening for breast and cervical cancer and elevated blood cholesterol. BRFSS data have been used to guide health promotion/disease prevention programs. REPORTING PERIOD: 1991. DESCRIPTION OF SYSTEM: BRFSS is a state-based random digit-dialing telephone survey of noninstitutionalized adults (> or = 18 years of age); 47 states and the District of Columbia participated in BRFSS in 1991. RESULTS: Some year 2000 objectives appear to be readily attainable for many states, whereas others do not. For example, among participating states, a median 57.8% (range = 45.6%-82.8%) of women ages > or = 50 years reported having had both a clinical breast examination and a mammogram in the previous 2 years (year 2000 objective: > or = 60%). In contrast, a median 37.3% (range = 22.1%-52.5%) of persons with annual family income < $20,000 reported that they did not engage in leisure-time physical activity--more than twice the year 2000 objective (> or = 17%). INTERPRETATION: BRFSS data demonstrate substantial state-to-state variation in progress toward year 2000 objectives and highlight areas (e.g., lack of leisure-time physical activity) in which substantial progress remains to be made in most states. Action taken: The BRFSS will continue to report data that relate to year 2000 health objectives. BRFSS data will enable states to monitor progress toward these objectives and develop health policies aimed at achieving them. PMID- 8413177 TI - Behavioral Risk Factor Surveillance System: summary of data for 1991. AB - PROBLEM/CONDITION: High-risk behaviors, such as smoking cigarettes and driving under the influence of alcohol, contribute heavily to morbidity and mortality from noninfectious disease and injury. Substantial variation exists among states in the prevalences of these behaviors. REPORTING PERIOD: 1991. DESCRIPTION OF SYSTEM: The Behavioral Risk Factor Surveillance System (BRFSS) is a state-based random-digit-dialing telephone survey of noninstitutionalized adults (> or = 18 years of age). In 1991, 47 states and the District of Columbia participated in BRFSS. The system focuses on behaviors that are related to one or more of the 10 leading causes of death. In 1991, BRFSS also began collecting data on self reported lack of health insurance. RESULTS: As in previous years, BRFSS data for 1991 indicate substantial state-to-state variation in the prevalence of risk factors such as chronic or binge alcohol consumption, sedentary lifestyle, and overweight. In addition to measures reported in previous years, the current report includes state prevalences of high blood cholesterol awareness (range = 13.5%-21.5%; median = 16.9%) and lack of health insurance (range = 7.2%-25.7%; median = 14.5%). INTERPRETATION: Because prevalence estimates vary considerably from state to state, state estimates may be preferable to national ones for use in planning programs. ACTIONS TAKEN: The BRFSS will continue to provide state specific data about health behaviors to allow states to monitor trends that affect the burden of chronic diseases in the United States. PMID- 8413178 TI - Specific chromosomal sites enhancing homologous recombination in Escherichia coli mutants defective in RNase H. AB - To clone new replication origin(s) activated under RNase H-defective (rnh-) conditions in Escherichia coli cells, whole chromosomal DNA digested with EcoRI was to with a Kmr DNA fragment and transformed into an rnh- derivative host. From the Kmr transformants, we obtained eight kinds of plasmid-like DNA, each of which contained a specific DNA fragment, termed "Hot", derived from the E. coli genome. Seven of the Hot DNAs (HotA-G) mapped to various sites within a narrow DNA replication termination region (about 280 kb), without any particular selection. Because Hot DNA could not be transformed into a mutant strain in which the corresponding Hot region had been deleted from the chromosome, the Hot DNA, though obtained as covalently closed circular (ccc) DNA, must have arisen by excision from the host chromosome into which it had initially integrated, rather than by autonomous replication of the transformed species. While Hot DNA does not have a weak replication origin it does have a strong recombinational hotspot active in the absence of RNase H. This notion is supported by the finding that Chi activity was present on all Hot DNAs tested and no Hot-positive clone without Chi activity was obtained, with the exception of a DNA clone carrying the dif site. PMID- 8413179 TI - Genetic analysis of human p34CDC2 function in fission yeast. AB - The p34cdc2 protein kinase plays a key role in the control of the mitotic cell cycle of fission yeast, being required for both entry into S-phase and for entry into mitosis in the mitotic cell cycle, as well as for the initiation of the second meiotic nuclear division. In recent years, structural and functional homologues of p34cdc2, as well as several of the proteins that interact with and regulate p34cdc2 function in fission yeast, have been identified in a wide range of higher eukaryotic cell types, suggesting that the control mechanisms uncovered in this simple eukaryote are likely to be well conserved across evolution. Here we describe the construction and characterisation of a fission yeast strain in which the endogenous p34cdc2 protein is entirely absent and is replaced by its human functional homologue p34CDC2. We have used this strain to analyse aspects of the function of the human p34CDC2 protein genetically. We show that the function of the human p34CDC2 protein in fission yeast cells is dependent upon the action of the protein tyrosine phosphatase p80cdc25, that it responds to altered levels of both the mitotic inhibitor p107wee1 and the p34cdc2-binding protein p13suc1, and is lethal in combination with the mutant B-type cyclin p56cdc13-117. In addition, we demonstrate that the human p34CDC2 protein is proficient for fission yeast meiosis, and examine the behaviour of two mutant p34CDC2 proteins in fission yeast. PMID- 8413180 TI - MSI3, a multicopy suppressor of mutants hyperactivated in the RAS-cAMP pathway, encodes a novel HSP70 protein of Saccharomyces cerevisiae. AB - The MSI3 gene was isolated as a multicopy suppressor of the heat shock-sensitive phenotype of the ira1 mutation, which causes hyperactivation of the RAS-cAMP pathway. Overexpression of MSI3 also suppresses the heat shock-sensitive phenotype of the bcy1 mutant. Determination of the DNA sequence of MSI3 revealed that MSI3 can encode a 77.4 kDa protein related to the HSP70 family. The amino acid sequence of Msi3p is about 30% identical to that of the Ssa1p of Saccharomyces cerevisiae. This contrasts with the finding that members of the HSP70 family generally show at least 50% amino acid identity. The consensus nucleotide sequence of the heat shock element (HSE) was found in the upstream region of MSI3. Moreover, the steady-state levels of the MSI3 mRNA and protein were increased upon heat shock. These results indicate that the MSI3 gene encodes a novel HSP70-like heat shock protein. Disruption of the MSI3 gene was associated with a temperature sensitive growth phenotype but unexpectedly, thermotolerance was enhanced in the disruptant. PMID- 8413181 TI - A heat shock-activated cDNA rescues the recessive lethality of mutations in the heterochromatin-associated protein HP1 of Drosophila melanogaster. AB - HP1 is a small nonhistone chromosomal protein of Drosophila melanogaster predominantly localized to the pericentric heterochromatin. We have shown previously that mutations in the HP1 coding sequences are associated with dominant suppression of heterochromatic position-effect variegation, and with recessive lethality. When fused to an Hsp70 heat shock gene promoter, the cDNA encoding HP1 supports the heat shock-inducible accumulation of HP1 protein in transgenic flies; this cDNA construct complements the dominant suppression of position-effect variegation associated with mutations in the HP1 gene. Here, we report experiments demonstrating that the heat shock-driven HP1 cDNA is capable of fully rescuing the recessive lethality associated with HP1 mutations in a heat shock-dependent fashion. If heat shock-induced HP1 expression is delayed for as long as 5 days, more than half of the mutant flies still survive until adulthood, consistent with a substantial maternal contribution to embryonic and larval viability. Elevating HP1 levels as late as 7-8 days of development is sufficient to enhance variegation three-fold, suggesting that the extent of heterochromatic position effect can be modified subsequent to the initial appearance of HP1 in the nuclei of syncytial blastoderm embryos. PMID- 8413182 TI - Osmoregulation of the fatty acid receptor gene fadL in Escherichia coli. AB - The fadL gene of Escherichia coli codes for an outer membrane protein that is involved in the uptake of long-chain fatty acids. Uptake is regulated by environmental osmolarity, and decreases when the cells are grown under conditions of high osmolarity. A temperature-sensitive mutant that requires fatty acid for growth at 42 degrees C was unable to grow at the high temperature even in the presence of fatty acid if the medium contained 10% sucrose. Promoter activity of the fadL gene in vivo was repressed by high osmolarity in a FadR repressor null mutant. Furthermore, in vitro transcription of the fadL gene was strongly repressed by the addition of OmpR and EnvZ proteins. The results of gel retardation and DNase I protection experiments indicated that OmpR, after incubation with the protein kinase EnvZ, specifically binds to at least four sites around the fadL promoter, two upstream and two downstream from the transcriptional start site. These results suggest that transcription of the fadL gene is osmotically regulated by the OmpR-EnvZ two-component system. PMID- 8413183 TI - Excess histidine enzymes cause AICAR-independent filamentation in Escherichia coli. AB - High-level expression of the hisHAFI genes in Escherichia coli, cloned under the control of an IPTG-inducible promoter, caused filamentation, as previously reported in Salmonella typhimurium. We speculated that this filamentation might be produced by an action of the HisH and HisF enzymes on their product AICAR (amino-imidazole carboxamide riboside 5'-phosphate), a histidine by-product and normal purine precursor, possibly by favouring the formation of ZTP, the triphosphate derivative of AICAR. However, filamentation occurred even in the absence of carbon flow through the histidine and purine pathways, as observed in a hisG purF strain lacking the first enzyme in each pathway. Filamentation thus does not require either the normal substrate or products of the overproduced histidine enzymes and must reflect another activity. PMID- 8413184 TI - AICAR is not an endogenous mutagen in Escherichia coli. AB - A number of observations in the Escherichia coli and Salmonella typhimurium literature could be explained by the hypothesis that a particular purine ribonucleotide precursor can be converted to the corresponding deoxyribonucleotide triphosphate, thereby becoming a base-analogue mutagen. The metabolite in question, AICAR (5-amino-4-carboxamide imidazole riboside 5' phosphate), is also a by-product of histidine biosynthesis, and its (ribo)triphosphate derivative, ZTP, has been detected in E. coli. We constructed E. coli tester strains that had either a normal AICAR pool (pur+ his+ strains cultivated without purines or histidine) or no AICAR pool (purF hisG mutant strains, lacking the first enzyme of each pathway and cultivated in the presence of adenine and histidine). Using a set of lacZ mutations, each of which can revert to Lac+ only by a specific substitution mutation, we found that no base substitution event occurs at a higher frequency in the presence of an AICAR pool. We conclude that the normal AICAR pool in E. coli is not a significant source of spontaneous base substitution mutagenesis. PMID- 8413185 TI - The pleiotropic effects of his overexpression in Salmonella typhimurium do not involve AICAR-induced mutagenesis. AB - Inhibition of cell division associated with overexpression of hisH and hisF in Salmonella typhimurium is strongly reminiscent of a cellular response to DNA damage. On these grounds, we investigated the involvement of a metabolite which appeared to represent a possible candidate for an endogenous mutagen: the base analog 5-amino-4-carboxamide imidazole riboside 5'-phosphate (AICAR), a by product of HisH and HisF activity. However, we showed that AICAR is not an endogenous mutagen in S. typhimurium. Other types of DNA damage induced by his overexpression seem also unlikely, since similar mutation rates are found in hisO+ and hisOc strains. We also show that AICAR production is not involved in the pleiotropic effects of his overexpression, since these are still observed in strains devoid of AICAR. Thus inhibition of cell division resulting from HisH and HisF overexpression must operate through a mechanism unrelated to the role of these proteins in histidine biosynthesis. PMID- 8413186 TI - A tobacco cDNA clone encoding a GATA-1 zinc finger protein homologous to regulators of nitrogen metabolism in fungi. AB - In higher plants, the expression of the nitrate assimilation pathway is highly regulated. Although the molecular mechanisms involved in this regulation are currently being elucidated, very little is known about the trans-acting factors that allow expression of the nitrate and nitrite reductase genes which code for the first enzymes in the pathway. In the fungus Neurospora crassa, nit-2, the major nitrogen regulatory gene, activates the expression of unlinked structural genes that specify nitrogen-catabolic enzymes during conditions of nitrogen limitation. The nit-2 gene encodes a regulatory protein containing a single zinc finger motif defined by the C-X2-C-X17-C-X2-C sequence. This DNA-binding domain recognizes the promoter region of N. crassa nitrogen-related genes and fragments derived from the tomato nia gene promoter. The observed specificity of the binding suggests the existence of a NIT2-like homolog in higher plants. PCR and cross-hybridization techniques were used to isolate, respectively, a partial cDNA from Nicotiana plumbaginifolia and a full-length cDNA from Nicotiana tabacum. These clones encode a NIT2-like protein (named NTL1 for nit-2-like), characterized by a single zinc finger domain, defined by the C-X2-C-X18-C-X2-C amino acids, and associated with a basic region. The amino acid sequence of NTL1 is 60% homologous to the NIT2 sequence in the zinc finger domain. The Ntl1 gene is present as a unique copy in the diploid N. plumbaginifolia species. The characteristics of Ntl1 gene expression are compatible with those of a regulator of the nitrate assimilation pathway, namely weak nitrate inducibility and regulation by light. PMID- 8413187 TI - The centromere and promoter factor, 1, CPF1, of Saccharomyces cerevisiae modulates gene activity through a family of factors including SPT21, RPD1 (SIN3), RPD3 and CCR4. AB - In Saccharomyces cerevisiae, the CPF1 gene encodes a centromere binding protein that also plays a role in transcription; cpf1 strains are methionine auxotrophs. In this paper we describe four strains that are methionine prototrophs despite containing a defective CPF1 gene. These strains, which contain mutations at either the SPT21, RPD1 (SIN3), RPD3 or CCR4 loci, have defective centromere function and a chromatin structure around the CDEI elements in the MET25 promoter characteristic of strains lacking CPF1. This indicates that the roles of CPF1 in transcription, centromere function and chromatin modulation around CDEI sites are different. We propose that CPF1 functions to overcome the repressing action, mediated via inactive chromatin, of proteins such as SPT21 or RPD1 (SIN3) on gene expression. The absence of proteins such as SPT21 or RPD1 (SIN3) relieves this repression and explains how methionine prototrophy is restored in the absence of CPF1. PMID- 8413188 TI - Five nitrate assimilation-related loci are clustered in Chlamydomonas reinhardtii. AB - Three overlapping clones covering a Chlamydomonas reinhardtii genomic region of about 32 kb appear to contain five genes potentially involved in nitrate assimilation in addition to the nitrate reductase structural locus nit-1. These new loci produced transcripts of 2.8, 2.2, 1.8 and 1.7 kb in nitrate-induced wild type cells that, like the 3.4 kb transcript of nit-1, were undetectable in cells grown in ammonium. In addition, in a mutant defective at the regulatory locus, nit-2 for nitrate assimilation, which does not express the nit-1 gene transcript, accumulation of the four other transcripts was also blocked. They have been named nar (nitrate assimilation related) genes. The nar-1 and nar-2 loci are transcribed in the same orientation as nit-1. The nar-3 and nar-4 loci are transcribed divergently from nit-1. DNA and RNA sequences from both nar-3 and nar 4 cross-hybridized with each other indicating that they share similar sequences. Four nitrate assimilation-deficient mutants (C2, D2, F6 and G1) were characterized. These mutants lack nar transcripts and have major deletions and/or rearrangements in the nar gene cluster. In contrast to other nitrate reductase deficient mutants and to wild type, deletion mutants and the regulatory mutant nit-2 were incapable of accumulating intracellular nitrate. Two of the mutants in which expression of all the nar loci did not occur, C2 and D2, grew in nitrite medium and showed wild-type levels of both nitrite uptake and nitrite reductase activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413189 TI - TdcA, a transcriptional activator of the tdcABC operon of Escherichia coli, is a member of the LysR family of proteins. AB - The tdcB and tdcC genes of the tdcABC operon of Escherichia coli encode threonine dehydratase and a threonine-serine permease, respectively. These proteins are involved in transport and metabolism of threonine and serine during anaerobic growth. In this study, we functionally characterized tdcA, which encodes a 35 kDa polypeptide consisting of 312 amino acid residues. Non-polar and partially polar mutations introduced into tdcA drastically reduced the expression of the genes down-stream from tdcA. Complementation studies using single-copy chromosomal integrants of a tdcB-lacZ fusion harboring an in-frame deletion of tdcA with chromosomal or plasmid-borne tdcA+ in trans showed complete restoration of tdc operon expression in vivo. The amino acid sequence at the amino-terminal end of TdcA revealed a significant homology to the helix-turn-helix motifs of typical DNA binding proteins. Sequence alignment of TdcA with LysR also showed considerable sequence similarity throughout their entire lengths. Our results suggest that TdcA is related to the LysR family of proteins by common ancestry and, based on its functional role in tdc expression, belongs to the LysR family of transcriptional activators. PMID- 8413190 TI - The atypical chlorophyll a/b/c light-harvesting complex of Mantoniella squamata: molecular cloning and sequence analysis. AB - cDNA species encoding precursor polypeptides of the chlorophyll a/b/c light harvesting complex (LHC) of Mantoniella squamata were cloned and sequenced. The precursor polypeptides have molecular weights of 24.2 kDa and are related to the major chlorophyll a/b polypeptides of higher plants. Southern analysis showed that their genes belong to the nuclear encoded Lhc multigene family; the investigated genes most probably do not contain introns. The chlorophyll a/b/c polypeptides contain two highly conserved regions common to all LHC polypeptides and three hydrophobic alpha-helices, which span the thylakoid membrane. The first membrane-spanning helix, however, is not detected by predictive methods: its atypical hydrophilic domains may bind the chlorophyll c molecules within the hydrophobic membrane environment. Homology to LHC II of higher plants and green algae is specifically evident in the C-terminal region comprising helix III and the preceding stroma-exposed domain. The N-terminal region of 29 amino acids resembles the structure of a transit sequence, which shows only minor similarities to those of LHC II sequences. Strikingly, the mature light harvesting polypeptides of M. squamata lack an N-terminal domain of 30 amino acids, which, in higher plants, contains the phosphorylation site of LHC II and simultaneously mediates membrane stacking. Therefore, the chlorophyll a/b/c polypeptides of M. squamata do not exhibit any light-dependent preference for photosystem I or II. The lack of this domain also indicates that the attractive forces between stacked thylakoids are weak. PMID- 8413191 TI - Localization of a cruciform cutting endonuclease to yeast mitochondria. AB - We have found a cruciform cutting endonuclease in the yeast, Saccharomyces cerevisiae, which localizes to the mitochondria. This activity apparently is associated with the mitochondrial inner membrane since the activity is not released into solution by osmolysis, in contrast to the matrix enzyme, isocitrate dehydrogenase. The cruciform cutting activity appears to be encoded by CCE1. This gene has been shown to encode one of the major cruciform cutting endonucleases present in yeast cell. In cce1 strains, which lack CCE1 endonuclease activity, the mitochondrial cruciform cutting endonucleolytic activity is also absent. Since CCE1 is allelic to MGT1, a gene required for the highly biased transmission of petite mitochondrial DNA in crosses between rho+ and hypersuppressive rho- cells, it seems likely that the CCE1 endonuclease functions within mitochondria. PMID- 8413192 TI - The NAM1/MTF2 nuclear gene product is selectively required for the stability and/or processing of mitochondrial transcripts of the atp6 and of the mosaic, cox1 and cytb genes in Saccharomyces cerevisiae. AB - The NAM1/MTF2 gene was firstly isolated as a multicopy suppressor of mitochondrial splicing deficiencies and independently as a gene of which a thermosensitive allele affects mitochondrial transcription in organello. To determine which step in mitochondrial RNA metabolism is controlled in vivo by the NAM1 gene, mitochondrial transcripts of seven transcription units from strains carrying an inactive nam1::URA3 gene disruption in various mitochondrial genetic backgrounds were analysed by Northern blot hybridisations. In a strain carrying an intron-containing mitochondrial genome, the inactivation of the NAM1 gene led to a strong decrease in (or total absence of) the mosaic cytb and cox1 mRNAs and in transcripts of the atp6-rf3/ens2 genes, which are co-transcribed with cox1. Neither the accumulation of unspliced cytb or cox1 pre-mRNAs, nor that of excised circular intron molecules of ai1 or ai2 were observed, but the abundance of the bi1 and ai7 lariats was comparable to that observed in the wild-type strain, thus demonstrating that transcription of the cytb and cox1 genes does occur. In strains carrying the intron-less mitochondrial genome with or without the rf3/ens2 sequence, wild-type amounts of cytb and cox1 mRNAs were detected while the amount of the atp6 mRNA was always strongly decreased. The abundance of transcripts from five other genes was either slightly (21S rRNA) or not at all (cox2, cox3, atp9 and 15S rRNA) affected by the nam1 inactivation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413193 TI - Functional analysis of the Lactococcus lactis usp45 secretion signal in the secretion of a homologous proteinase and a heterologous alpha-amylase. AB - The ups45 gene encodes the major extracellular protein from Lactococcus lactis. The deduced sequence of the 27 residue leader peptide revealed the tripartite characteristics of a signal peptide. This leader peptide directed the efficient secretion of the homologous proteinase (PrtP) in L. lactis, indicating that the putative signal peptide of PrtP can be replaced by the 27 residue Usp45 leader peptide. In addition, the 27 residue leader peptide could be used to secrete the Bacillus stearothermophilus alpha-amylase, encoded by the amyS gene. Fusion of the usp45 promoter region and various parts of the leader sequence to an amyS gene devoid of its signal sequence, showed that in Escherichia coli the first 19, 20, and 27 residues of the Usp45 leader are able to direct alpha-amylase secretion. In L. lactis the shorter signal peptides did not result in secretion of alpha-amylase, providing experimental evidence for the hypothesis that gram positive bacteria require a longer signal peptide for secretion than gram negative organisms. PMID- 8413194 TI - Genes at different regulatory levels are required for the ammonia control of nodulation in Rhizobium meliloti. AB - The expression of the nodulation genes nodABC of Rhizobium meliloti, which determine early response functions to plant host signals, is regulated by the level of ammonia, the primary product of symbiotic nitrogen fixation. We show that the pathway that links the ammonia-induced signal to the transcriptional control of the nodABC genes involves at least two regulatory levels. The fluctuating nitrogen level is sensed and the signal is mediated by the members of the general nitrogen regulatory (ntr) system, then transmitted to the syrM-nodD3 genes representing the nod-specific level of ammonia regulation. At low ammonia concentration, the activator protein NtrC exerts its effect via nodD3. In conditions of nitrogen excess ntrR, involved in the repression of nod genes, may function in coordination with the syrM gene. Finally, the NodD3 protein may relay the nitrogen status signal to the transcriptional control of the nodABC genes. PMID- 8413195 TI - Ribosomal protein gene rpl5 is cotranscribed with the nad3 gene in Oenothera mitochondria. AB - The rpl5 ribosomal protein gene was identified in the mitochondrial genome of the higher plant Oenothera berteriana. The gene is present in a unique genomic location upstream of the gene encoding subunit 3 of the NADH dehydrogenase (nad3). Both genes are cotranscribed, and the mRNA is modified at several cytidine residues by RNA editing. Analysis of the editing profiles of both genes by direct cDNA analysis and polymerase chain reaction (PCR) revealed that not all transcripts are fully edited at all sites. Eight of the nine C to U conversions in the rpl5 reading frame are non-silent and change the deduced amino acid sequence. The genes of the prokaryotic-like cistron that includes the rpsl9, rps3, rpl16, rpl5, and rpsl4 genes, which is at least partially conserved in the mitochondrial genomes of other higher and lower plants, are dispersed in the Oenothera mitochondrial genome. PMID- 8413196 TI - Photolyase-dimer-DNA complexes and exclusion stimulation in Escherichia coli: depolarization of the plasma membrane. AB - Using cells that overproduce DNA photolyase, we found that UV irradiation (3 J/m2) efficiently inactivates accumulation of methylthiogalactoside (TMG) when RexAB proteins of phage lambda are present. The effect requires both formation of photolyase-dimer-DNA (PDD) complexes and expression of the RexAB proteins. It is reversed completely by a flash of visible light if given immediately after UV and becomes irreversible after post-UV incubation for about 15 min. Inactivation is significant after only 5 min of post-UV incubation, is accompanied by a loss of previously accumulated TMG, and does not require de novo protein synthesis. Passive transport of O-nitrophenylgalactoside by inactivated cells is typical of energy-depleted membranes. We suggest that PDD complexes mimic a developmental intermediate of phage superinfection and stimulate formation of the RexB membrane channel recently proposed by others to explain classical "exclusion". This suggestion is supported by additional data showing an inactivation of colony forming ability by exclusion stimulation and an inability of PDD complexes to inactivate accumulation of TMG if RexB is present in larger relative amounts than RexA (a detail characteristic of exclusion stimulated by phage superinfection). PMID- 8413197 TI - Evidence from directed mutagenesis that positively charged amino acids are necessary for interaction of nitrogenase with the [2Fe-2S] heterocyst ferredoxin (FdxH) from the cyanobacterium Anabaena sp., PCC7120. AB - Sequence comparison of the heterocyst-type ferredoxin (FdxH) from Anabaena 7120 and type-1 ferredoxins (PetF) from the same organism and other cyanobacteria revealed a group of positively charged residues characteristic for FdxH. Molecular modeling showed that these basic amino acids are clustered on the surface of FdxH. The corresponding domain of PetF contained acidic or nonpolar residues instead. To identify amino acids that are important for interaction with nitrogenase, we generated site-directed mutations in the fdxH gene and assayed the in vitro activity of the resulting recombinant proteins isolated from Escherichia coli. In addition to the point mutants, two chimeric proteins, FdxH:PetF and PetF:FdxH, were constructed containing the 58 N-terminal amino acids of one ferredoxin fused to the 40 C-terminal amino acids of the other. Exchange of lysines 10 and 11 of FdxH for the corresponding residues of PetF (glutamate 10 and alanine 11) resulted in a ferredoxin with greatly decreased affinity to nitrogenase. This indicates an important function of these basic amino acids in interaction with dinitrogenase reductase (NifH) from Anabaena. In addition we checked the reactivity of the recombinant ferredoxins with ferredoxin NADP+ oxidoreductase (FNR) and photosystem I. The experiments with both the chimeric and point mutated ferredoxins showed that the C-terminal part of this protein determines its activity in NADP+ photoreduction. PMID- 8413198 TI - Analysis of a replication initiation sequence from the adenosine deaminase region of the mouse genome. AB - A 4-kb HindIII fragment that supported the efficient autonomous replication of plasmid vector pDY-, a replication-defective construct based on Epstein-Barr virus sequences, in human K562 cells was rescued from amplified double-minute chromosomes containing the murine adenosine deaminase locus. Polymerase chain reaction assays of size-fractionated nascent strands demonstrated that replication initiation occurred within the same 1- to 2-kb region of this fragment in autonomously replicating plasmids containing the sequence in either orientation, in double-minute chromosomes, and in the single-copy locus at its normal chromosomal location. The complete sequence of this fragment was determined; it contains a 248-bp polypurine tract and consensus binding site sequences for several putative transcription and replication factors. PMID- 8413199 TI - Fibroblast growth factor inhibits MRF4 activity independently of the phosphorylation status of a conserved threonine residue within the DNA-binding domain. AB - MRF4 is a member of the muscle-specific basic helix-loop-helix transcription factor family that also includes MyoD, myogenin, and Myf-5. Each of these proteins, when overexpressed in fibroblasts, converts the cells to differentiated muscle fibers that express several skeletal muscle genes, such as those for alpha actin, muscle creatine kinase, and troponin I. Despite the fact that MRF4 functions as a positive transcriptional regulator, the MRF4 protein is subject to negative regulation by a variety of agents, most notably by exposure of cells to purified growth factors, such as basic fibroblast growth factor (bFGF). In an effort to establish whether bFGF inhibits MRF4 activity through specific posttranslational modifications, we examined whether MRF4 exists in vivo as a phosphoprotein and whether the phosphorylation status of the protein regulates its activity. Our results indicate that MRF4 is phosphorylated predominantly on serine residues, with weak phosphorylation occurring on threonine residues. Both cyclic AMP-dependent protein kinase (PKA) and protein kinase C (PKC) phosphorylate MRF4 in vitro as well as in vivo, and the overexpression of each kinase inhibits MRF4 activity and thus blocks terminal differentiation. PKC directed phosphorylation of a conserved threonine residue (T-99) situated within the DNA-binding domain inhibits MRF4 from binding in vitro to specific DNA targets. However, although T-99 itself is essential for myogenic activity, our studies demonstrate that the phosphorylation status of T-99 does not play a major role in regulating MRF4 activity in vivo, since PKA, PKC, and bFGF inhibit the activity of MRF4 proteins in which the identified PKA and PKC sites have been mutated. We suggest that the negative regulation of MRF4 imposed by bFGF does not involve a direct modification of the protein at the identified PKA and PKC sites but instead may involve the modification of specific coregulators that interact with this muscle regulatory factor. PMID- 8413200 TI - Pi, a pre-B-cell-specific enhancer element in the immunoglobulin heavy-chain enhancer. AB - We have identified a new immunoglobulin heavy-chain enhancer element, designated pi, between the microE2 and microE3 elements. The pi enhancer element is transcriptionally active primarily during early stages of B-cell development but becomes virtually inactive during B-cell maturation at about the stage of immunoglobulin kappa light-chain gene rearrangement. Mutational analysis suggests that the pi element is crucial for immunoglobulin heavy-chain enhancer activity at the pre-B-cell stage but is almost irrelevant for enhancer activity at the mature B-cell or plasma-cell stage. The activity of the pi enhancer element correlates with the presence of an apparently pre-B-cell-specific protein-DNA complex. The similarity of the pi site to recognition sequences for members of the ets gene family suggests that the protein(s) interacting with the pi site most likely are ets-related transcription factors. PMID- 8413201 TI - ADA3: a gene, identified by resistance to GAL4-VP16, with properties similar to and different from those of ADA2. AB - We describe the isolation of a yeast gene, ADA3, mutations in which prevent the toxicity of GAL4-VP16 in vivo. Toxicity was previously proposed to be due to the trapping of general transcription factors required at RNA polymerase II promoters (S. L. Berger, B. Pina, N. Silverman, G. A. Marcus, J. Agapite, J. L. Regier, S. J. Triezenberg, and L. Guarente, Cell 70:251-265, 1992). trans activation by VP16 as well as the acidic activation domain of GCN4 is reduced in the mutant. Other activation domains, such as those of GAL4 and HAP4, are only slightly affected in the mutant. This spectrum is similar to that observed for mutants with lesions in ADA2, a gene proposed to encode a transcriptional adaptor. The ADA3 gene is not absolutely essential for cell growth, but gene disruption mutants grow slowly and are temperature sensitive. Strains doubly disrupted for ada2 and ada3 grow no more slowly than single mutants, providing further evidence that these genes function in the same pathway. Selection of initiation sites by the general transcriptional machinery in vitro is altered in the ada3 mutant, providing a clue that ADA3 could be a novel general transcription factor involved in the response to acidic activators. PMID- 8413202 TI - A U3 small nuclear ribonucleoprotein-requiring processing event in the 5' external transcribed spacer of Xenopus precursor rRNA. AB - A processing site has been identified within the 5' external transcribed spacer (ETS) of Xenopus laevis and X. borealis pre-RNAs, and this in vivo processing can be reproduced in vitro. It involves a stable and specific association of the pre rRNA with factors in the cell extract, including at least four RNA-contacting polypeptides, yielding a distinct complex that sediments at 20S. Processing also requires the U3 small nuclear RNA. This processing, at residue +105 of the 713 nucleotide X. laevis 5' ETS, is highly reminiscent of the initial processing cleavage of mouse pre-rRNA within its 3.5-kb 5' ETS, previously thought to be mammal specific. The frog and mouse processing signals share a short essential sequence motif, and mouse factors can faithfully process the frog pre-rRNA. This conservation suggests that this 5' ETS processing site serves an evolutionarily selective function. PMID- 8413203 TI - Control of calcitonin/calcitonin gene-related peptide pre-mRNA processing by constitutive intron and exon elements. AB - The calcitonin/calcitonin gene-related peptide (CGRP) primary transcript is alternatively spliced in thyroid C cells and neurons, resulting in the tissue specific production of calcitonin and CGRP mRNAs. Analyses of mutated calcitonin/CGRP transcription units in permanently transfected cell lines have indicated that alternative splicing is regulated by a differential capacity to utilize the calcitonin-specific splice acceptor. The analysis of an extensive series of mutations suggests that tissue-specific regulation of calcitonin mRNA production does not depend on the presence of a single, unique cis-active element but instead appears to be a consequence of suboptimal constitutive splicing signals. While only those mutations that altered constitutive splicing signals affected splice choices, the action of multiple regulatory sequences cannot be formally excluded. Further, we have identified a 13-nucleotide purine-rich element from a constitutive exon that, when placed in exon 4, entirely switches splice site usage in CGRP-producing cells. These data suggest that specific exon recruitment sequences, in combination with other constitutive elements, serve an important function in exon recognition. These results are consistent with the hypothesis that tissue-specific alternative splicing of the calcitonin/CGRP primary transcript is mediated by cell-specific differences in components of the constitutive splicing machinery. PMID- 8413204 TI - Dominant missense mutations in a novel yeast protein related to mammalian phosphatidylinositol 3-kinase and VPS34 abrogate rapamycin cytotoxicity. AB - Rapamycin is a macrolide antifungal agent that exhibits potent immunosuppressive properties. In Saccharomyces cerevisiae, rapamycin sensitivity is mediated by a specific cytoplasmic receptor which is a homolog of human FKBP12 (hFKBP12). Deletion of the gene for yeast FKBP12 (RBP1) results in recessive drug resistance, and expression of hFKBP12 restores rapamycin sensitivity. These data support the idea that FKBP12 and rapamycin form a toxic complex that corrupts the function of other cellular proteins. To identify such proteins, we isolated dominant rapamycin-resistant mutants both in wild-type haploid and diploid cells and in haploid rbp1::URA3 cells engineered to express hFKBP12. Genetic analysis indicated that the dominant mutations are nonallelic to mutations in RBP1 and define two genes, designated DRR1 and DRR2 (for dominant rapamycin resistance). Mutant copies of DRR1 and DRR2 were cloned from genomic YCp50 libraries by their ability to confer drug resistance in wild-type cells. DNA sequence analysis of a mutant drr1 allele revealed a long open reading frame predicting a novel 2470 amino-acid protein with several motifs suggesting an involvement in intracellular signal transduction, including a leucine zipper near the N terminus, two putative DNA-binding sequences, and a domain that exhibits significant sequence similarity to the 110-kDa catalytic subunit of both yeast (VPS34) and bovine phosphatidylinositol 3-kinases. Genomic disruption of DRR1 in a mutant haploid strain restored drug sensitivity and demonstrated that the gene encodes a nonessential function. DNA sequence comparison of seven independent drr1dom alleles identified single base pair substitutions in the same codon within the phosphatidylinositol 3-kinase domain, resulting in a change of Ser-1972 to Arg or Asn. We conclude either that DRR1 (alone or in combination with DRR2) acts as a target of FKBP12-rapamycin complexes or that a missense mutation in DRR1 allows it to compensate for the function of the normal drug target. PMID- 8413205 TI - Alternative splicing of Pax-8 gene transcripts is developmentally regulated and generates isoforms with different transactivation properties. AB - Pax-8, a member of the paired box-containing gene family, was shown to be coexpressed with Pax-2 in several human kidney carcinoma cell lines. Four different Pax-8 mRNA isoforms, a to d, were cloned from one of these cell lines by polymerase chain reaction amplification, and the Pax-8 gene was isolated from a human cosmid library. Analysis of the exon-intron structure of Pax-8 revealed that the four mRNA isoforms arise by alternative splicing, resulting in inclusion or exclusion of exon 7 and/or exon 8 sequences. All four Pax-8 proteins retain the paired domain as their DNA-binding motif and recognize DNA in the same manner as do the closely related Pax-2 and BSAP (Pax-5) proteins. The Pax-8a and Pax-8b isoforms end in a serine/threonine/tyrosine-rich sequence, while the C terminus of Pax-8c and Pax-8d is translated in a different, proline-rich reading frame. Transient transfection experiments revealed that Pax-8 isoforms a and b, but not c and d, strongly stimulate transcription from a promoter containing six copies of a paired-domain recognition sequence. The same four mRNA variants were also detected by RNase protection analysis in the mouse embryo and adult kidney, thus indicating evolutionary conservation of Pax-8 mRNA splicing. A different splice pattern was observed in the developing placenta, which expresses two new variants, Pax-8e and Pax-8f, instead of transcripts b to d. Expression of these mRNAs is high at embryonic day 9.5 and is gradually reduced until Pax-8a is the predominant transcript in the 12.5-day placenta. In the embryo, however, the synthesis of mRNAs b to d is initially low and then increases relative to that of Pax-8a. Hence, alternative splicing of Pax-8 gene transcripts not only generates six different Pax-8 variants but is also temporally and spatially regulated during early mouse development. PMID- 8413206 TI - Expression of bovine myf5 induces ectopic skeletal muscle formation in transgenic mice. AB - myf5 is one of a family of four myogenic determination genes that control skeletal muscle differentiation. To study the role of myf5 in vivo, we generated transgenic mice harboring the bovine homolog, bmyf, under control of the murine sarcoma virus promoter. Ectopic expression of the full-length bmyf transgene was detected in brain and heart tissue samples of F1 progeny from transgenic founder mice. Ectopic bmyf expression activated endogenous skeletal myogenic determination genes in the hearts and brains of transgenic animals. Incomplete skeletal myogenesis in most hearts gave rise to cardiomegaly and focal areas of cardiomyopathy. In brains in which ectopic expression led to a more complete myogenesis, focal areas of multinucleated, striated myotubes containing actin, desmin, and myosin were observed. These unexpected results show that myf5 can initiate myogenic differentiation in vivo, supporting the hypothesis that myf5 is responsible for determination of cells to the myogenic lineage in normal embryogenesis. PMID- 8413207 TI - Internalization of activated platelet-derived growth factor receptor phosphatidylinositol-3' kinase complexes: potential interactions with the microtubule cytoskeleton. AB - Phosphatidylinositol (PI)-3' kinase catalyzes the formation of PI 3,4-diphosphate and PI 3,4,5-triphosphate in response to stimulation of cells by platelet-derived growth factor (PDGF). Here we report that tyrosine-phosphorylated PDGF receptors, the p85 subunit of PI-3' kinase (p85), and activated PI-3' kinase are found in isolated clathrin-coated vesicles within 2 min of exposure of cells to PDGF, indicating that both receptor and activated PI-3' kinase enter the endocytic pathway. Immunofluorescence analysis of p85 in serum-starved cells revealed a punctate/reticular staining pattern, concentrated in the perinuclear region and displaying high focal concentration at the centrosome. In addition, partial coalignment of p85 with microtubules was observed after optical sectioning microscopy and image reconstruction. The association of p85 with the microtubule network was further evidenced by the microtubule-depolymerizing drug nocodazole, which caused a redistribution of p85 from the perinuclear region to the cell periphery. Interestingly, the most significant effect of PDGF on the distribution of p85 was an increase in the staining intensity of this protein in the perinuclear region, and this effect was eliminated by prior treatment of cells with nocodazole. These results suggest that PDGF receptor-p85 complexes internalize and transit in association with the microtubule cytoskeleton. In addition, the high concentration of p85 in intracellular structures in the absence of PDGF stimulation suggests additional roles for this protein independent of its association with receptor tyrosine kinases. PMID- 8413208 TI - Generation of a new adenovirus type 12-inducible fragile site by insertion of an artificial U2 locus in the human genome. AB - Infection with adenovirus type 12 (Ad12) induces four fragile sites in the human genome (H.F. Stich, G.L. van Hoosier, and J.J. Trentin, Exp. Cell Res. 34:400 403, 1964; H. zur Hausen, J. Virol. 1:1174-1185, 1967). The major site, at 17q21 22, contains the U2 gene cluster, which is specifically disrupted by infection in at least a percentage of the cells (D.M. Durnam, J.C. Menninger, S.H. Chandler, P.P. Smith, and J.K. McDougall, Mol. Cell. Biol. 8:1863-1867, 1988). For direct assessment of whether the U2 locus is the target of the Ad12 effect, an artificial locus, constructed in vitro and consisting of tandem arrays of the U2 6-kbp monomer, was transfected into human cells. We report that integration of this artificial locus on the p arm of chromosome 13 creates a new Ad12-inducible fragile site. PMID- 8413209 TI - The Rox1 repressor of the Saccharomyces cerevisiae hypoxic genes is a specific DNA-binding protein with a high-mobility-group motif. AB - The ROX1 gene encodes a repressor of the hypoxic functions of the yeast Saccharomyces cerevisiae. The DNA sequence of the gene was determined and found to encode a protein of 368 amino acids. The amino-terminal third of the protein contains a high-mobility-group motif characteristic of DNA-binding proteins. To determine whether the Rox1 repressor bound DNA, the gene was expressed in Escherichia coli cells as a fusion to the maltose-binding protein and this fusion was partially purified by amylose affinity chromatography. By using a gel retardation assay, both the fusion protein and Rox1 itself were found to bind specifically to a synthetic 32-bp DNA containing the hypoxic consensus sequence. We assessed the role of the general repressor Ssn6 in ANB1 repression. An ANB1 lacZ fusion was expressed constitutively in an ssn6 deletion strain, and deletion of the Rox1 binding sites in the ANB1 upstream region did not increase the level of derepression, suggesting that Ssn6 exerts its effect through Rox1. Finally, ROX1 was mapped to yeast chromosome XVI, near the ARO7-OSM2 locus. PMID- 8413210 TI - Retinoic acid repression of cell-specific helix-loop-helix-octamer activation of the calcitonin/calcitonin gene-related peptide enhancer. AB - We have investigated the mechanism underlying repression of calcitonin/calcitonin gene-related peptide (CT/CGRP) gene expression by retinoic acid. Retinoic acid treatment of the CA77 thyroid C-cell line decreased CT/CGRP promoter activity two to threefold, which correlates well with the decrease in calcitonin and CGRP mRNA levels. Repression is mediated through the nuclear retinoic acid receptors (RAR) on the basis of the retinoid specificity, the sensitivity of repression (half-maximal repression at 0.2 nM), and the additional repression caused by cotransfection of an alpha-RAR expression vector. The sequences required for retinoic acid repression were localized to an 18-bp element containing cell specific enhancer activity. The enhancer binds helix-loop-helix (HLH) and octamer transcription factors that act synergistically to activate transcription. Retinoic acid repression requires both these factors since mutations in either motif resulted in the loss of repression. Furthermore, repression was observed only in cell lines containing enhancer activity. We have used electrophoretic mobility shift assays to show that repression does not involve direct DNA binding of RAR or RAR-retinoid X receptor heterodimers. Instead, repression appears to involve interactions with the stimulatory enhancer factors. Following retinoic acid treatment, there was a specific decrease in an enhancer complex containing both HLH and octamer proteins. Formation of the HLH-octamer complex was also specifically blocked by the addition of exogenous RAR-retinoid X receptor protein. These results demonstrate that RAR can repress CT/CGRP gene transcription by interfering with combinatorial activation by cell-specific HLH and octamer proteins. PMID- 8413211 TI - NF-kappa B p100 (Lyt-10) is a component of H2TF1 and can function as an I kappa B like molecule. AB - NF-kappa B is an important transcription factor regulating expression of genes involved in immune function, inflammation, and cellular growth control. NF-kappa B activity is induced by numerous stimuli, such as phorbol esters, B- and T-cell mitogens, the cytokines tumor necrosis factor and interleukin-1, and serum growth factors. The standard model for the induction of NF-kappa B activity involves the release of the transcription factor from a cytoplasmic inhibitor termed I kappa B, allowing translocation of NF-kappa B to the nucleus. I kappa B contains multiple copies of the so-called ankyrin repeat, which are apparently necessary for its function. Subunits comprising NF-kappa B and related binding activities are members of the Rel multigene family. Two such subunits, p50 and p52 (also called p50B), are proteolytically processed from precursors of 105 kDa (also called p105 and NFKB1) and 100 kDa (also called p100, NFKB2, and Lyt-10), respectively. Both contain N-terminal Rel-homologous domains as well as multiple copies of C-terminal ankyrin repeats. We show here that NF-kappa B p100 is a component of the previously identified DNA-binding activity H2TF1. In addition, we show that p100 is localized in the cytoplasm in HeLa cells, where it is associated with c-Rel, p50, or p65 (RelA). In transient-transfection assays, p100 represses the ability of NF-kappa B p65 to activate a kappa B-containing reporter construct. Transfection of p100 also results in a loss of nuclear p65 DNA binding to a kappa B probe, as measured by an electrophoretic mobility shift assay, and a loss of nuclear p65 immunoreactivity, as measured by immunoblotting. This loss of nuclear p65 is paralleled by a gain of p65 DNA-binding activity and immunoreactivity in the cytoplasm. We interpret these data as demonstrating that p100 functions as an I kappa B-like molecule to sequester Rel family members in the cytoplasm. Proteolytic processing of p100 to the activator p52 is predicted to generate several new forms of Rel family heterodimers and therefore represents a form of regulation of NF-kappa B activity distinct from the classic I kappa B pathway. PMID- 8413212 TI - PUB1 is a major nuclear and cytoplasmic polyadenylated RNA-binding protein in Saccharomyces cerevisiae. AB - Proteins that directly associate with nuclear polyadenylated RNAs, or heterogeneous nuclear RNA-binding proteins (hnRNPs), and those that associate with cytoplasmic mRNAs, or mRNA-binding proteins (mRNPs), play important roles in regulating gene expression at the posttranscriptional level. Previous work with a variety of eukaryotic cells has demonstrated that hnRNPs are localized predominantly within the nucleus whereas mRNPs are cytoplasmic. While studying proteins associated with polyadenylated RNAs in Saccharomyces cerevisiae, we discovered an abundant polyuridylate-binding protein, PUB1, which appears to be both an hnRNP and an mRNP. PUB1 and PAB1, the polyadenylate tail-binding protein, are the two major proteins cross-linked by UV light to polyadenylated RNAs in vivo. The deduced primary structure of PUB1 indicates that it is a member of the ribonucleoprotein consensus sequence family of RNA-binding proteins and is structurally related to the human hnRNP M proteins. Even though the PUB1 protein is a major cellular polyadenylated RNA-binding protein, it is nonessential for cell growth. Indirect cellular immunofluorescence combined with digital image processing allowed a detailed comparison of the intracellular distributions of PUB1 and PAB1. While PAB1 is predominantly, and relatively uniformly, distributed within the cytoplasm, PUB1 is localized in a nonuniform pattern throughout both the nucleus and the cytoplasm. The cytoplasmic distribution of PUB1 is considerably more discontinuous than that of PAB1. Furthermore, sucrose gradient sedimentation analysis demonstrates that PAB1 cofractionates with polyribosomes whereas PUB1 does not. These results suggest that PUB1 is both an hnRNP and an mRNP and that it may be stably bound to a translationally inactive subpopulation of mRNAs within the cytoplasm. PMID- 8413213 TI - PUB1: a major yeast poly(A)+ RNA-binding protein. AB - The expression of RNA polymerase II transcripts can be regulated at the posttranscriptional level by RNA-binding proteins. Although extensively characterized in metazoans, relatively few RNA-binding proteins have been characterized in the yeast Saccharomyces cerevisiae. Three major proteins are cross-linked by UV light to poly(A)+ RNA in living S. cerevisiae cells. These are the 72-kDa poly(A)-binding protein and proteins of 60 and 50 kDa (S.A. Adam, T.Y. Nakagawa, M.S. Swanson, T. Woodruff, and G. Dreyfuss, Mol. Cell. Biol. 6:2932 2943, 1986). Here, we describe the 60-kDa protein, one of the major poly(A)+ RNA binding proteins in S. cerevisiae. This protein, PUB1 [for poly(U)-binding protein 1], was purified by affinity chromatography on immobilized poly(rU), and specific monoclonal antibodies to it were produced. UV cross-linking demonstrated that PUB1 is bound to poly(A)+ RNA (mRNA or pre-mRNA) in living cells, and it was detected primarily in the cytoplasm by indirect immunofluorescence. The gene for PUB1 was cloned and sequenced, and the sequence was found to predict a 51-kDa protein with three ribonucleoprotein consensus RNA-binding domains and three glutamine- and asparagine-rich auxiliary domains. This overall structure is remarkably similar to the structures of the Drosophila melanogaster elav gene product, the human neuronal antigen HuD, and the cytolytic lymphocyte protein TIA 1. Each of these proteins has an important role in development and differentiation, potentially by affecting RNA processing. PUB1 was found to be nonessential in S. cerevisiae by gene replacement; however, further genetic analysis should reveal important features of this class of RNA-binding proteins. PMID- 8413214 TI - Activation of the inducible orphan receptor gene nur77 by serum growth factors: dissociation of immediate-early and delayed-early responses. AB - We have characterized the genetic elements that mediate the transcriptional activation of nur77, a growth factor-inducible gene encoding a member of the steroid/thyroid hormone receptor superfamily. Although initially identified as a serum-inducible immediate-early gene with expression kinetics similar to those of c-fos, we found that transcriptional activation of nur77 by serum growth factors in fibroblasts is in fact composed of two components: an immediate-early component, which can occur in the absence of de novo protein synthesis, and a delayed-early component, which is dependent on de novo protein synthesis. The expression of nur77 following serum stimulation reflects the superimposition of immediate-early and delayed-early expression. Immediate-early and delayed-early expression can be dissociated from one another by deletion or base substitution mutations of the nur77 promoter. Immediate-early expression of nur77 is mediated primarily by sequences located between nucleotides -86 and -126 upstream of the transcription start site. This region includes a sequence that resembles but differs from the CArG element found in other serum-inducible promoters. Upstream of the CArG-like element is a potential binding site for a transcription factor of the Ets family; the presence of this site is required for significant transcriptional induction. Delayed-early expression of nur77 is mediated by multiple AP-1-like and GC-rich elements, which can interact with products of immediate-early genes such as Fos/Jun and Zif268, respectively. Furthermore, we show that Zif268 can activate transcription of the nur77 promoter, suggesting that it may play a role in the delayed-early expression of nur77. PMID- 8413215 TI - NF-kappa B subunit-specific regulation of the interleukin-8 promoter. AB - Interleukin-8 (IL-8), a chemotactic cytokine for T lymphocytes and neutrophils, is induced in several cell types by a variety of stimuli including the inflammatory cytokines IL-1 and tumor necrosis factor alpha TNF-alpha. Several cis elements, including a binding site for the inducible transcription factor NF kappa B, have been identified in the regulatory region of the IL-8 gene. We have examined the ability of various NF-kappa B subunits to bind to, and activate transcription from, the IL-8 promoter. A nuclear complex was induced in phorbol myristate acetate-treated Jurkat T cells which bound specifically to the kappa B site of the IL-8 promoter and was inhibited by addition of purified I kappa B alpha to the reaction mixture. Only antibody to RelA (p65), but not to NFKB1 (p50), NFKB2 (p50B), c-Rel, or RelB was able to abolish binding, suggesting that RelA is a major component in these kappa B binding complexes. Gel mobility shift analysis with in vitro-translated and purified proteins indicated that whereas the kappa B element in the human immunodeficiency virus type 1 long terminal repeat bound to all members of the kappa B/Rel family examined, the IL-8 kappa B site bound only to RelA and to c-Rel and NFKB2 homodimers, but not to NFKB1 homodimers or heterodimers of NFKB1-RelA. Transient transfection analysis demonstrated a kappa B-dependent expression of the IL-8 promoter in a human fibrosarcoma cell line (8387) and in Jurkat T lymphocytes. Cotransfection with various NF-kappa B subunits indicated that RelA and c-Rel, but neither NFKB1 nor heterodimeric NFKB1-RelA, was able to activate transcription from the IL-8 promoter. Furthermore, cotransfection of NFKB1 and RelA, although able to support activation from the human immunodeficiency virus type 1 long terminal repeat, failed to activate expression from the IL-8 promoter. Antisense oligonucleotides to RelA, but not NFKB1, inhibited phorbol myristate acetate-induced IL-8 production in Jurkat T lymphocytes. These data demonstrate the differential ability of members of the kappa B/Rel family to bind to, and activate transcription from, the IL-8 promoter. Furthermore, while providing a novel example of a kappa B-regulated promoter in which the classical NF-kappa B complex is unable to activate transcription from the kappa B element, these data provide direct evidence for the role of RelA in regulation of IL-8 gene expression. PMID- 8413216 TI - Identification of a rel-related protein in the nucleus during the S phase of the cell cycle. AB - The c-rel proto-oncogene encodes a 75-kDa protein (p75c-rel) which is present in the cytosol of chick embryo fibroblasts (CEF) associated with a distinct set of cellular proteins with molecular masses of 40, 115, and 124 kDa. CEF cultures arrested in S phase of the cell cycle, or enriched for G2 or mitotic cells, were examined to determine whether the expression of c-rel was altered during the cell cycle. Levels of p75c-rel remained constant in all portions of the cell cycle examined; however, a Rel-related protein with an apparent molecular mass of 64 kDa was detected in nuclei of S-phase cells. As cells enter G2, the level of this protein in the nucleus decreases. This protein reacts with antiserum generated against the carboxy terminus of p75c-rel in radioimmunoprecipitations and Western immunoblot experiments and was also detected in a Western immunoblot with antiserum generated against the first 161 amino acids of pp59v-rel. Thus, unlike other Rel/NF-kappa B family members, p64 has carboxy-terminal homology with c Rel. The majority of peptides generated by partial proteolytic cleavage of p64 are shared with peptides generated by digestion of p75c-rel and/or pp59v-rel. We suggest that this protein represents a new member of the Rel family of transcription factors and is located in the nucleus of avian fibroblasts during S phase of the cell cycle. PMID- 8413217 TI - Retinoic acid induction of major histocompatibility complex class I genes in NTera-2 embryonal carcinoma cells involves induction of NF-kappa B (p50-p65) and retinoic acid receptor beta-retinoid X receptor beta heterodimers. AB - Retinoic acid (RA) treatment of human embryonal carcinoma (EC) NTera-2 (NT2) cells induces expression of major histocompatibility complex (MHC) class I and beta-2 microglobulin surface molecules. We found that this induction was accompanied by increased levels of MHC class I mRNA, which was attributable to the activation of the two conserved upstream enhancers, region I (NF-kappa B like) and region II. This activation coincided with the induction of nuclear factor binding activities specific for the two enhancers. Region I binding activity was not present in undifferentiated NT2 cells, but binding of an NF kappa B heterodimer, p50-p65, was induced following RA treatment. The p50-p65 heterodimer was produced as a result of de novo induction of p50 and p65 mRNAs. Region II binding activity was present in undifferentiated cells at low levels but was greatly augmented by RA treatment because of activation of a nuclear hormone receptor heterodimer composed of the retinoid X receptor (RXR beta) and the RA receptor (RAR beta). The RXR beta-RAR beta heterodimer also bound RA responsive elements present in other genes which are likely to be involved in RA triggering of EC cell differentiation. Furthermore, transfection of p50 and p65 into undifferentiated NT2 cells synergistically activated region I-dependent MHC class I reporter activity. A similar increase in MHC class I reporter activity was demonstrated by cotransfection of RXR beta and RAR beta. These data show that following RA treatment, heterodimers of two transcription factor families are induced to bind to the MHC enhancers, which at least partly accounts for RA induction of MHC class I expression in NT2 EC cells. PMID- 8413218 TI - Nuclear dot antigens may specify transcriptional domains in the nucleus. AB - A bank of 892 human autoimmune serum samples was screened by indirect immunofluorescence on human tissue culture HT-29 cells. Seven serum samples that stain 4 to 10 bright dots in cell lines of several different mammals, including humans, monkeys, rats, and pigs, were identified. Immunofluorescence experiments indicate that these antigens, called nuclear dot (ND) antigens, are distinct from splicing complexes, kinetochores, and other known nuclear structures. An ND antigen recognized by these sera was cloned by immunoscreening a human lambda gt11 expression library. Analysis of seven cDNA clones for the ND antigen indicates that several mRNAs exist, perhaps derived through alternative splicing mechanisms. One major form of the message has an open reading frame of 1,440 bp capable of encoding a 53,000-M(r) protein. Treatment of cells with detergent, salt, or RNase A fails to remove the ND antigen from the nucleus. However, incubation with DNase I obliterates ND staining, indicating that the ND protein directly or indirectly associates with nuclear DNA. Fusion of the ND protein to a LexA DNA binding domain activates transcription in Saccharomyces cerevisiae. A 75 amino-acid domain that activates transcription in both yeast and primate cells has been identified. We suggest that ND antigens may participate in the activation of transcription of specific regions of the genome. PMID- 8413219 TI - Integration of simian virus 40 into cellular DNA occurs at or near topoisomerase II cleavage hot spots induced by VM-26 (teniposide). AB - Inhibition of DNA topoisomerase II in simian virus 40 (SV40)-infected BSC-1 cells with a topoisomerase II poison, VM-26 (teniposide), resulted in rapid conversion of a population of the SV40 DNA into a high-molecular-weight form. Characterization of this high-molecular-weight form of SV40 DNA suggests that it is linear, double stranded, and a recombinant with SV40 DNA sequences covalently joined to cellular DNA. The majority of the integrants contain fewer than two tandem copies of SV40 DNA. Neither DNA-damaging agents, such as mitomycin and UV, nor the topoisomerase I inhibitor camptothecin induced detectable integration in this system. In addition, the recombination junctions within the SV40 portion of the integrants correlate with VM-26-induced, topoisomerase II cleavage hot spots on SV40 DNA. These results suggest a direct and specific role for topoisomerase II and possibly the enzyme-inhibitor-DNA ternary cleavable complex in integration. The propensity of poisoned topoisomerase II to induce viral integration also suggests a role for topoisomerase II in a pathway of chromosomal DNA rearrangements. PMID- 8413220 TI - Characterization of the human interleukin-2 receptor beta-chain gene promoter: regulation of promoter activity by ets gene products. AB - The interleukin-2 receptor (IL-2R) beta chain (IL-2R beta) is an essential signaling component of high- and intermediate-affinity IL-2Rs. Our laboratory previously reported that a DNA fragment containing 857 bp of 5'-flanking sequence of the human IL-2R beta gene exhibited promoter activity. We have now further characterized the promoter and delineated cis-acting regulatory regions. The region downstream of -363 is critical for basal and phorbol myristate acetate inducible IL-2R beta promoter activity and contains at least three enhancer-like regions. Among them, the -56 to -34 enhancer was the most potent and had high level activity in two T-cell lines but not in nonlymphoid HeLaS3 and MG63 cells. This enhancer contains a GGAA Ets binding site which bound two Ets family proteins, Ets-1 and GA-binding protein in vitro. Mutation of the Ets motif strongly diminished both promoter and enhancer activities. We conclude that this Ets binding site plays a key role in regulating basal and phorbol myristate acetate-inducible IL-2R beta promoter activity and may also contribute to tissue specific expression of the IL-2R beta gene. PMID- 8413221 TI - Order of intron removal during splicing of endogenous adenine phosphoribosyltransferase and dihydrofolate reductase pre-mRNA. AB - Using a strategy based on reverse transcription and the polymerase chain reaction, we have determined the order of splicing of the four introns of the endogenous adenine phosphoribosyltransferase (aprt) gene in Chinese hamster ovary cells. The method involves a pairwise comparison of molecules that retain one intron and have either retained or spliced another intron(s). A highly preferred order of removal was found: intron 3 > 2 > 4 = 1. This order did not represent a linear progression from one end of the transcript to the other, nor did it correlate with the conformity of the splice site sequences to the consensus sequences or to the calculated energy of duplex formation with U1 small nuclear RNA. By using actinomycin D to inhibit RNA synthesis, the in vivo rate of the first step in splicing was estimated for all four introns; a half-life of 6 min was found for introns 2, 3, and 4. Intron 1 was spliced more slowly, with a 12 min half-life. A substantial amount of RNA that retained intron 1 as the sole intron was exported to the cytoplasm. In the course of these experiments, we also determined that intron 3, but not intron 4, is spliced before 3'-end formation is complete, probably on nascent transcripts. This result is consistent with the idea that polyadenylation is required for splicing of the 3'-most intron. We applied a similar strategy to determine the last intron to be spliced in a very large transcript, that of the endogenous dihydrofolate reductase (dhfr) gene in Chinese hamster ovary cells (25 kb). Here again, intron 1 was the last intron to be spliced. PMID- 8413222 TI - A V(D)J recombinase-inducible B-cell line: role of transcriptional enhancer elements in directing V(D)J recombination. AB - Rapid analysis of mechanisms that regulate V(D)J recombination has been hampered by the lack of appropriate cell systems that reproduce aspects of normal prelymphocyte physiology in which the recombinase is activated, accessible antigen receptor loci are rearranged, and rearrangement status is fixed by termination of recombinase expression. To generate such a system, we introduced heat shock-inducible V(D)J recombination-activating genes (RAG) 1 and 2 into a recombinationally inert B-cell line. Heat shock treatment of these cells rapidly induced high levels of RAG transcripts and RAG proteins that were accompanied by a parallel induction of V(D)J recombinase activity, strongly suggesting that RAG proteins have a primary role in V(D)J recombination. Within hours after induction, these cells began to rearrange chromosomally integrated V(D)J recombination substrates but only if the substrates contained an active transcriptional enhancer; substrates lacking an enhancer were not efficiently rearranged. Activities necessary to target integrated substrates for rearrangement were provided by two separate lymphoid-specific transcriptional enhancers, as well as an active nonlymphoid enhancer, unequivocally demonstrating that such elements enhance both transcription and V(D)J recombinational accessibility. PMID- 8413223 TI - Characterization of a functional NF-kappa B site in the human interleukin 1 beta promoter: evidence for a positive autoregulatory loop. AB - The -300 region of the interleukin 1 beta (IL-1 beta) promoter contains a functional NF-kappa B binding site composed of the decamer sequence 5'-GGGAAAATCC 3'. Probes representing the -300 region or the NF-kappa B site alone interacted with NF-kappa B proteins present in phorbol myristate acetate-, lipopolysaccharide-, or Sendai virus-induced myeloid cell extracts as well as recombinant NFKB1 (p50) and RelA (p65); furthermore, NF-kappa B protein-DNA complex formation was dissociated in vitro by the addition of recombinant I kappa B alpha. Mutation of the NF-kappa B site in the context of the IL-1 beta promoter reduced the responsiveness of the IL-1 beta promoter to various inducers, including phorbol ester, Sendai virus, poly(rI-rC), and IL-1 beta. A 4.4-kb IL-1 beta promoter fragment linked to a chloramphenicol acetyltransferase reporter gene was also preferentially inducible by coexpression of individual NF-kappa B subunits compared with a mutated IL-1 beta promoter fragment. When multiple copies of the IL-1 beta NF-kappa B site were linked to an enhancerless simian virus 40 promoter, this element was able to mediate phorbol ester- or lipopolysaccharide-inducible gene expression. In cotransfection experiments, RelA (p65) and c-Rel (p85) were identified as the main subunits responsible for the activation of the IL-1 beta NF-kappa B site; also, combinations of NFKB1 (p50) and RelA (p65) or c-Rel and RelA were strong transcriptional activators of reporter gene activity. The presence of a functional NF-kappa B binding site in the IL-1 beta promoter suggests that IL-1 positively autoregulates its own synthesis, since IL-1 is a strong inducer of NF-kappa B binding activity. Thus, the IL-1 beta gene may be considered as an important additional member of the family of cytokine genes regulated in part by the NF-kappa B/rel family of transcription factors. PMID- 8413224 TI - Conditional transformation of cells and rapid activation of the mitogen-activated protein kinase cascade by an estradiol-dependent human raf-1 protein kinase. AB - We report a strategy for regulating the activity of a cytoplasmic signaling molecule, the protein kinase encoded by raf-1. Retroviruses encoding a gene fusion between an oncogenic form of human p74raf-1 and the hormone-binding domain of the human estrogen receptor (hrafER) were constructed. The fusion protein was nontransforming in the absence of estradiol but could be reversibly activated by the addition or removal of estradiol from the growth media. Activation of hrafER was accompanied in C7 3T3 cells by the rapid, protein synthesis-independent activation of both mitogen-activated protein (MAP) kinase kinase and p42/p44 MAP kinase and by phosphorylation of the resident p74raf-1 protein as demonstrated by decreased electrophoretic mobility. The phosphorylation of p74raf-1 had no effect on the kinase activity of the protein, indicating that mobility shift is an unreliable indicator of p74raf-1 enzymatic activity. Removal of estradiol from the growth media led to a rapid inactivation of the MAP kinase cascade. These results demonstrate that Raf-1 can activate the MAP kinase cascade in vivo, independent of other "upstream" signaling components. Parallel experiments performed with rat1a cells conditionally transformed by hrafER demonstrated activation of MAP kinase kinase in response to estradiol but no subsequent activation of p42/p44 MAP kinases or phosphorylation of p74raf-1. This result suggests that in rat1a cells, p42/p44 MAP kinase activation is not required for Raf-1-mediated oncogenic transformation. Estradiol-dependent activation of p42/p44 MAP kinases and phosphorylation of p74raf-1 was, however, observed in rat1a cells expressing hrafER when the cells were pretreated with okadaic acid. This result suggests that the level of protein phosphatase activity may play a crucial role in the regulation of the MAP kinase cascade. Our results provide the first example of a cytosolic signal transducer being harnessed by fusion to the hormone-binding domain of the estrogen receptor. This conditional system not only will aid the elucidation of the function of Raf-1 but also may be more broadly useful for the construction of conditional forms of other kinases and signaling molecules. PMID- 8413225 TI - Potential RNA polymerase II-induced interactions of transcription factor TFIIB. AB - The ubiquitous transcription factor TFIIB is required for initiation by RNA polymerase II and serves as a target of some regulatory factors. The carboxy terminal portion of TFIIB contains a large imperfect direct repeat reminiscent of the structural organization of the TATA-binding component (TBP) of TFIID, as well as sequence homology to conserved regions of bacterial sigma factors. The present study shows that the carboxy-terminal portion of TFIIB, like that of TBP, is folded into a compact protease-resistant core. The TFIIB core, unlike the TBP core, is inactive in transcription but retains structural features that enable it to form a complex with promoter-bound TFIID. The protease-susceptible amino terminus appears to contain components responsible for direct interaction with RNA polymerase II (in association with TFIIF) either on the promoter (in association with TFIID) or independently. In addition, core TFIIB (but not intact TFIIB) extends the footprint of TBP on promoter DNA, suggesting that TFIIB has a cryptic DNA-binding potential. These results are consistent with a model in which TFIIB, in a manner functionally analogous to that of bacterial sigma factors, undergoes an RNA polymerase II-dependent conformational change with resultant DNA interactions during the pathway leading to a functional preinitiation complex. PMID- 8413226 TI - A growth factor-induced kinase phosphorylates the serum response factor at a site that regulates its DNA-binding activity. AB - A signaling pathway by which growth factors may induce transcription of the c-fos proto-oncogene has been characterized. Growth factor stimulation of quiescent fibroblasts activates a protein kinase cascade that leads to the rapid and transient phosphorylation of the serum response factor (SRF), a regulator of c fos transcription. The in vivo kinetics of SRF phosphorylation and dephosphorylation parallel the activation and subsequent repression of c-fos transcription, suggesting that this phosphorylation event plays a critical role in the control of c-fos expression. The ribosomal S6 kinase pp90rsk, a growth factor-inducible kinase, phosphorylates SRF in vitro at serine 103, the site that becomes newly phosphorylated upon growth factor stimulation in vivo. Phosphorylation of serine 103 significantly enhances the affinity and rate with which SRF associates with its binding site, the serum response element, within the c-fos promoter. These results suggest a model in which the growth factor induced phosphorylation of SRF at serine 103 contributes to the activation of c fos transcription by facilitating the formation of an active transcription complex at the serum response element. PMID- 8413227 TI - Connections between the Ras-cyclic AMP pathway and G1 cyclin expression in the budding yeast Saccharomyces cerevisiae. AB - We have identified two processes in the G1 phase of the Saccharomyces cerevisiae cell cycle that are required before nutritionally arrested cells are able to return to proliferative growth. The first process requires protein synthesis and is associated with increased expression of the G1 cyclin gene CLN3. This process requires nutrients but is independent of Ras and cyclic AMP (cAMP). The second process requires cAMP. This second process is rapid, is independent of protein synthesis, and produces a rapid induction of START-specific transcripts, including CLN1 and CLN2. The ability of a nutritionally arrested cell to respond to cAMP is dependent on completion of the first process, and this is delayed in cells carrying a CLN3 deletion. Mating pheromone blocks the cAMP response but does not alter the process upstream of Ras-cAMP. These results suggest a model linking the Ras-cAMP pathway with regulation of G1 cyclin expression. PMID- 8413228 TI - Transcriptional up-regulation of the mouse cytosolic glutathione peroxidase gene in erythroid cells is due to a tissue-specific 3' enhancer containing functionally important CACC/GT motifs and binding sites for GATA and Ets transcription factors. AB - Nuclear run-on experiments have shown that the high level of expression of the mouse cytosolic glutathione peroxidase mRNA in erythroid cells is due to up regulation of the gene at the transcriptional level. Studies of the chromatin structure around the cytosolic glutathione peroxidase gene have revealed a series of DNase I hypersensitive sites (DHSS) in the 3' flanking region of the gene in erythroid and other high-expression tissues that are lacking in low-expression cells, in addition to a DHSS over the promoter region in both high- and low expression tissues. Functional transfection experiments have demonstrated that one of the 3' DHSS regions functions as an enhancer in erythroid cells but not in a low-expression epithelial cell line; and site-directed mutagenesis and footprinting experiments reveal that the activity of the erythroid cell-specific enhancer requires a cluster of binding sites for the CACC/GT box factors and the GATA and Ets families of transcription factors. PMID- 8413229 TI - HSP78 encodes a yeast mitochondrial heat shock protein in the Clp family of ATP dependent proteases. AB - The Saccharomyces cerevisiae nuclear gene for a 78-kDa mitochondrial heat shock protein (hsp78) was identified in a lambda gt11 expression library through immunological screening with an hsp78-specific monoclonal antibody. Sequencing of HSP78 revealed a long open reading frame capable of encoding an 811-amino-acid, 91.3-kDa basic protein with a putative mitochondrial leader sequence and two potential nucleotide-binding sites. Sequence comparisons revealed that hsp78 is a member of the highly conserved family of Clp proteins and is most closely related to the Escherichia coli ClpB protein, which is thought to be an ATPase subunit of an intracellular ATP-dependent protease. The steady-state levels of HSP78 transcripts and protein varied in response to both thermal stress and carbon source with an approximately 30-fold difference between repressed levels in cells growing fermentatively on glucose at 30 degrees C and derepressed levels in heat shocked cells growing on a nonfermentable carbon source. The response to heat shock is consistent with the presence of a characteristic heat shock regulatory element in the 5'-flanking DNA. Submitochondrial fractionation showed that hsp78 is a soluble protein located in the mitochondrial matrix. Cells carrying disrupted copies of HSP78 lacked hsp78 but were not impaired in respiratory growth at normal and elevated temperatures or in the ability to survive and retain mitochondrial function after thermal stress. The absence of a strong mitochondrial phenotype in hsp78 mutants is comparable to the nonlethal phenotypes of mutations in other Clp genes in bacteria and yeast. HSP78 is the third gene, with SSC1 and HSP60, known to encode a yeast mitochondrial heat shock protein and the second gene, with HSP104, for a yeast ClpB homolog. PMID- 8413230 TI - A genetic analysis of the E2F1 gene distinguishes regulation by Rb, p107, and adenovirus E4. AB - The cellular transcription factor E2F appears to be a target for the regulatory action of the retinoblastoma tumor suppressor gene product. The recent isolation of the E2F1 cDNA clone, which encodes a polypeptide with properties characteristic of E2F, has now allowed a more detailed analysis of the regulation of E2F function by Rb as well as the Rb-related p107 protein and the adenovirus 19-kDa E4 gene product. Previous experiments have shown that each of these regulatory proteins can modulate the activity of cellular E2F. We find that each of these regulatory events can be mediated through the E2F1 product. Moreover, an examination of various E2F1 mutations reveals distinct specificities for these regulatory proteins. For instance, the ability of E4 to alter E2F1 function is dependent upon sequences within a putative leucine repeat of E2F1 as well as within the C-terminal acidic domain. In contrast, the leucine repeat element was not important for Rb- or p107-mediated inhibition of E2F1 activity. Although the C-terminal acidic domain of E2F1, previously shown to be important for Rb binding, appears to be a site for regulation of E2F1 by Rb and p107, point mutations within this region distinguish recognition by Rb and p107. These results underscore the complexity of E2F regulatory interactions and also demonstrate a qualitative distinction in the interactions of Rb and p107 with E2F1, perhaps reflective of functional differences. PMID- 8413231 TI - The stringency and magnitude of androgen-specific gene activation are combinatorial functions of receptor and nonreceptor binding site sequences. AB - The mechanism by which specific hormonal regulation of gene expression is attained in vivo is a paradox in that several of the steroid receptors recognize the same DNA element in vitro. We have characterized a complex enhancer of the mouse sex-limited protein (Slp) gene that is activated exclusively by androgens but not by glucocorticoids in transfection. Potent androgen induction requires both the consensus hormone response element (HRE) and auxiliary elements residing within the 120-bp DNA fragment C' delta 9. Multiple nonreceptor factors are involved in androgen specificity, with respect to both the elevation of androgen receptor activity and the inactivity of glucocorticoid receptor (GR), since clustered base changes at any of several sites reduce or abolish androgen induction and do not increase glucocorticoid response. However, moving the HRE as little as 10 bases away from the rest of the enhancer allows GR to function, suggesting that GR is repressed by juxtaposition to particular factors within the androgen-specific complex. Surprisingly, some sequence variations of the HRE itself, within the context of C' delta 9, alter the stringency of specificity, as well as the magnitude, of hormonal response. These HRE sequence effects on expression correspond in a qualitative manner with receptor binding, i.e., GR shows a threefold difference in affinities for HREs amongst which androgen receptor does not discriminate. Altering the HRE orientation within the enhancer also affects hormonal stringency, increasing glucocorticoid but not androgen response. The effect of these subtle variations suggests that they alter receptor position with respect to other factors. Thus, protein-protein interactions that elicit specific gene regulation are established by the array of DNA elements in a complex enhancer and can be modulated by sequence variations within these elements that may influence selection of precise protein contacts. PMID- 8413232 TI - Identification of AML-1 and the (8;21) translocation protein (AML-1/ETO) as sequence-specific DNA-binding proteins: the runt homology domain is required for DNA binding and protein-protein interactions. AB - The AML1 gene on chromosome 21 is disrupted in the (8;21)(q22;q22) translocation associated with acute myelogenous leukemia and encodes a protein with a central 118-amino-acid domain with 69% homology to the Drosophila pair-rule gene, runt. We demonstrate that AML-1 is a DNA-binding protein which specifically interacts with a sequence belonging to the group of enhancer core motifs, TGT/cGGT. Electrophoretic mobility shift analysis of cell extracts identified two AML-1 containing protein-DNA complexes whose electrophoretic mobilities were slower than those of complexes formed with AML-1 produced in vitro. Mixing of in vitro produced AML-1 with cell extracts prior to gel mobility shift analysis resulted in the formation of higher-order complexes. Deletion mutagenesis of AML-1 revealed that the runt homology domain mediates both sequence-specific DNA binding and protein-protein interactions. The hybrid product, AML-1/ETO, which results from the (8;21) translocation and retains the runt homology domain, both recognizes the AML-1 consensus sequence and interacts with other cellular proteins. PMID- 8413233 TI - The yeast KRE9 gene encodes an O glycoprotein involved in cell surface beta glucan assembly. AB - The yeast KRE9 gene encodes a 30-kDa secretory pathway protein involved in the synthesis of cell wall (1-->6)-beta-glucan. Disruption of KRE9 leads to serious growth impairment and an altered cell wall containing less than 20% of the wild type amount of (1-->6)-beta-glucan. Analysis of the glucan material remaining in a kre9 delta null mutant indicated a polymer with a reduced average molecular mass. kre9 delta null mutants also displayed several additional cell-wall-related phenotypes, including an aberrant multiply budded morphology, a mating defect, and a failure to form projections in the presence of alpha-factor. Double mutants were generated by crossing kre9 delta strains with strains harboring a null mutation in the KRE1, KRE6, or KRE11 gene, and each of these double mutants was found to be inviable in the SEY6210 background. Similar crosses with null mutations in the KRE5 and SKN1 genes indicated that these double mutants were no more severely affected than kre5 delta or kre9 delta single mutants alone. Antibodies were generated against Kre9p and detected an O glycoprotein of approximately 55 to 60 kDa found in the extracellular medium of a strain overproducing Kre9p. PMID- 8413234 TI - The Drosophila Polycomb-group gene Enhancer of zeste contains a region with sequence similarity to trithorax. AB - As is typical of Polycomb-group loci, the Enhancer of zeste [E(z)] gene negatively regulates the segment identity genes of the Antennapedia (ANT-C) and Bithorax (BX-C) gene complexes. A second class of loci, collectively known as the trithorax group, plays an antagonistic role as positive regulators of the ANT-C and BX-C genes. Molecular analysis of the E(z) gene predicts a 760-amino-acid protein product. A region of 116 amino acids near the E(z) carboxy terminus is 41.2% identical (68.4% similar) with a carboxy-terminal region of the trithorax protein. This portion of the trithorax protein is part of a larger region previously shown to share extensive homology with a human protein (ALL-1/Hrx) that is implicated in acute leukemias. Over this same 116 amino acids, E(z) and ALL-1/Hrx are 43.9% identical (68.4% similar). Otherwise, E(z) is not significantly similar to any previously described proteins. As this region of sequence similarity is shared by two proteins with antagonistic functions, we suggest that it may comprise a domain that interacts with a common target, either nucleic acid or protein. Opposite effects on transcription might then be determined by other portions of the two proteins. PMID- 8413235 TI - Molecular cloning of a human cDNA encoding a novel protein, DAD1, whose defect causes apoptotic cell death in hamster BHK21 cells. AB - The tsBN7 cell line, one of the mutant lines temperature sensitive for growth which have been isolated from the BHK21 cell line, was found to die by apoptosis following a shift to the nonpermissive temperature. The induced apoptosis was inhibited by a protein synthesis inhibitor, cycloheximide, but not by the bcl-2 encoded protein. By DNA-mediated gene transfer, we cloned a cDNA that complements the tsBN7 mutation. It encodes a novel hydrophobic protein, designated DAD1, which is well conserved (100% identical amino acids between humans and hamsters). By comparing the base sequences of the parental BHK21 and tsBN7 DAD1 cDNAs, we found that the DAD1-encoding gene is mutated in tsBN7 cells. The DAD1 protein disappeared in tsBN7 cells following a shift to the nonpermissive temperature, suggesting that loss of the DAD1 protein triggers apoptosis. PMID- 8413236 TI - RFX1 is identical to enhancer factor C and functions as a transactivator of the hepatitis B virus enhancer. AB - Hepatitis B virus gene expression is to a large extent under the control of enhancer I (EnhI). The activity of EnhI is strictly dependent on the enhancer factor C (EF-C) site, an inverted repeat that is bound by a ubiquitous nuclear protein known as EF-C. Here we report the unexpected finding that EF-C is in fact identical to RFX1, a novel transcription factor previously cloned by virtue of its affinity for the HLA class II X-box promoter element. This finding has allowed us to provide direct evidence that RFX1 (EF-C) is crucial for EnhI function in HepG2 hepatoma cells; RFX1-specific antisense oligonucleotides appear to inhibit EnhI-driven expression of the hepatitis B virus major surface antigen gene, and in transfection assays, RFX1 behaves as a potent transactivator of EnhI. Interestingly, transactivation of EnhI by RFX1 (EF-C) is not observed in cell lines that are not of liver origin, suggesting that the ubiquitous RFX1 protein cooperates with liver-specific factors. PMID- 8413237 TI - Palmitylation of an amino-terminal cysteine motif of protein tyrosine kinases p56lck and p59fyn mediates interaction with glycosyl-phosphatidylinositol anchored proteins. AB - Cross-linking of glycosyl-phosphatidylinositol (GPI)-anchored membrane proteins on T cells can trigger cell activation. We and others have shown an association between GPI-anchored proteins and the protein tyrosine kinases (PTKs) p56lck and p59fyn, suggesting a pathway for signaling through GPI-anchored proteins. Studies of decay-accelerating factor (DAF) or CD59 in either the C32 cell line or the HeLa cell line transfected with PTK cDNA demonstrated that the GPI-anchored proteins associated noncovalently with p56lck and p59fyn but not with p60src. Nonmyristylated versions of p56lck and p59fyn also failed to associate with the GPI-anchored proteins. Mutational analysis of the PTK demonstrated that the association with the GPI-anchored proteins mapped to the unique amino-terminal domains of the PTK. A chimeric PTK consisting of the 10 amino-terminal residues of p56lck or p59fyn replacing the corresponding amino acids in p60src was sufficient for association with DAF, but the converse constructs containing the first 10 amino acids of p60src plus the remainder of p56lck or p59fyn did not associate with DAF. Mutation of cysteine to serine at positions 3 and 6 in p59fyn or positions 3 and 5 in p56lck abolished the association of these kinases with DAF. Mutation of serine to cysteine at positions 3 and 6 in p60src conferred on p60src the ability to associate with DAF. Direct labeling with [3H]palmitate demonstrated palmitylation of this amino-terminal cysteine motif in p56lck. Thus, palmitylation of the amino-terminal cysteine residue(s) together with myristylation of the amino-terminal glycine residue defines important motifs for the association of PTKs with GPI-anchored proteins. PMID- 8413238 TI - Human ERCC5 cDNA-cosmid complementation for excision repair and bipartite amino acid domains conserved with RAD proteins of Saccharomyces cerevisiae and Schizosaccharomyces pombe. AB - Several human genes related to DNA excision repair (ER) have been isolated via ER cross-species complementation (ERCC) of UV-sensitive CHO cells. We have now isolated and characterized cDNAs for the human ERCC5 gene that complement CHO UV135 cells. The ERCC5 mRNA size is about 4.6 kb. Our available cDNA clones are partial length, and no single clone was active for UV135 complementation. When cDNAs were mixed pairwise with a cosmid clone containing an overlapping 5'-end segment of the ERCC5 gene, DNA transfer produced UV-resistant colonies with 60 to 95% correction of UV resistance relative to either a genomic ERCC5 DNA transformant or the CHO AA8 progenitor cells. cDNA-cosmid transformants regained intermediate levels (20 to 45%) of ER-dependent reactivation of a UV-damaged pSVCATgpt reporter plasmid. Our evidence strongly implicates an in situ recombination mechanism in cDNA-cosmid complementation for ER. The complete deduced amino acid sequence of ERCC5 was reconstructed from several cDNA clones encoding a predicted protein of 1,186 amino acids. The ERCC5 protein has extensive sequence similarities, in bipartite domains A and B, to products of RAD repair genes of two yeasts, Saccharomyces cerevisiae RAD2 and Schizosaccharomyces pombe rad13. Sequence, structural, and functional data taken together indicate that ERCC5 and its relatives are probable functional homologs. A second locus represented by S. cerevisiae YKL510 and S. pombe rad2 genes is structurally distinct from the ERCC5 locus but retains vestigial A and B domain similarities. Our analyses suggest that ERCC5 is a nuclear-localized protein with one or more highly conserved helix-loop-helix segments within domains A and B. PMID- 8413239 TI - Activity of chimeric U small nuclear RNA (snRNA)/mRNA genes in transfected protoplasts of Nicotiana plumbaginifolia: U snRNA 3'-end formation and transcription initiation can occur independently in plants. AB - Formation of the 3' ends of RNA polymerase II (Pol II)-specific U small nuclear RNAs (U snRNAs) in vertebrate cells is dependent upon transcription initiation from the U snRNA gene promoter. Moreover, U snRNA promoters are unable to direct the synthesis of functional polyadenylated mRNAs. In this work, we have investigated whether U snRNA 3'-end formation and transcription initiation are also coupled in plants. We have first characterized the requirements for 3'-end formation of an Arabidopsis U2 snRNA expressed in transfected protoplasts of Nicotiana plumbaginifolia. We found that the 3'-end-adjacent sequence CA (N)3 10AGTNNAA, conserved in plant Pol II-specific U snRNA genes, is essential for the 3'-end formation of U2 transcripts and, similar to the vertebrate 3' box, is highly tolerant to mutation. The 3'-flanking regions of an Arabidopsis U5 and a maize U2 snRNA gene can effectively substitute for the Arabidopsis U2 3'-end formation signal, indicating that these signals are functionally equivalent among different Pol II-transcribed snRNA genes. The plant U snRNA 3'-end formation signal can be recognized irrespective of whether transcription initiation occurs at U snRNA or mRNA gene promoters, although efficiency of 3' box utilization is higher when transcription initiation occurs at the U snRNA promoter. Moreover, transcripts initiated from the U2 gene promoter can be spliced and polyadenylated. Transcription from a Pol III-specific plant U snRNA gene promoter is not compatible with polyadenylation. Finally, we reveal that initiation at a Pol II-specific plant U snRNA gene promoter can occur in the absence of the snRNA coding region and a functional snRNA 3'-end formation signal, demonstrating that these sequences play no role in determining the RNA polymerase specificity of plant U snRNA genes. PMID- 8413240 TI - Elements and factors involved in tissue-specific and embryonic expression of the liver transcription factor LFB1 in Xenopus laevis. AB - LFB1 (HNF1) is a tissue-specific transcription factor found in the livers, stomachs, intestines, and kidneys of vertebrates. By analyzing the promoter of the Xenopus LFB1 gene, we identified potential autoregulation by LFB1 and regulation by HNF4, a transcription factor with a tissue distribution similar to that of LFB1. Injection of LFB1 promoter-chloramphenicol acetyltransferase constructs into Xenopus eggs revealed embryonic activation that is restricted to the region of the developing larvae expressing endogeneous LFB1. Proper embryonic activation was also observed with a rat LFB1 promoter. Deletion analysis of the Xenopus and rat promoters revealed that in both promoters embryonic activation is absolutely dependnet on the presence of an element that contains CCNCTCTC as the core consensus sequence. Since this element is recognized by the maternal factor OZ-1 previously described by N. Ovsenek, A. M. Zorn, and P. A. Krieg (Development 115:649-655, 1992), we might have identified the main constituents of a hierarchy that leads via LFB1 to the activation of tissue-specific genes during embryogenesis. PMID- 8413241 TI - Schizosaccharomyces pombe Spk1 is a tyrosine-phosphorylated protein functionally related to Xenopus mitogen-activated protein kinase. AB - Mitogen-activated protein kinase (MAPK) and its direct activator, MAPK kinase (MAPKK), have been suggested to play a pivotal role in a variety of signal transduction pathways in higher eukaryotes. The fission yeast Schizosaccharomyces pombe carries a gene, named spk1, whose product is structurally related to vertebrate MAPK. Here we show that Spk1 is functionally related to Xenopus MAPK. (i) Xenopus MAPK partially complemented a defect in the spk1- mutant. An spk1- diploid strain could not sporulate, but one carrying Xenopus MAPK could. (ii) Both Spk1 and Xenopus MAPK interfered with sporulation if overexpressed in S. pombe cells. (iii) Spk1 underwent tyrosine phosphorylation as does Xenopus MAPK. Tyrosine phosphorylation of Spk1 appeared to be dependent upon mating signals because it occurred in homothallic cells but not in heterothallic cells. Furthermore, this phosphorylation was diminished in a byr1 disruptant strain, suggesting that spk1 lies downstream of byr1, which encodes a MAPKK homolog in S. pombe. Taken together, the MAPKK-MAPK cascade may be evolutionarily conserved in signaling pathways in yeasts and vertebrates. PMID- 8413242 TI - Transmembrane topology of the mammalian KDEL receptor. AB - The mammalian KDEL receptor is an integral membrane protein with seven hydrophobic regions. Fusion proteins comprising a 37-kDa N-glycosylation reporter fused downstream of amino-terminal fragments of the KDEL receptor with varying numbers of hydrophobic regions were synthesized in an in vitro translation system containing canine pancreatic microsomes. The luminal or cytosolic orientation of the reporter, and hence of the hydrophilic region to which it is fused, was inferred from the presence or absence of glycosylation, which occurs only in the lumen of the microsomes. The cytosolic orientation of the N and C termini was also confirmed immunocytochemically. Our results suggest that the KDEL receptor is inserted into the membrane with only six transmembrane domains and that both the amino and carboxy termini are located in the cytoplasm. PMID- 8413243 TI - CYC2 encodes a factor involved in mitochondrial import of yeast cytochrome c. AB - The gene CYC2 from the yeast Saccharomyces cerevisiae was previously shown to affect levels of mitochondrial cytochrome c by acting at a posttranslational step in cytochrome c biosynthesis. We report here the cloning and identification of the CYC2 gene product as a protein involved in import of cytochrome c into mitochondria. CYC2 encodes a 168-amino-acid open reading frame with at least two potential transmembrane segments. Antibodies against a synthetic peptide corresponding to the carboxyl terminus of the predicted sequence were raised. These antibodies recognize multiple bands on immunoblots of mitochondrial extracts. The intensities of these bands vary according to the gene dosage of CYC2 in various isogenic strains. Immunoblotting of subcellular fractions suggests that the CYC2 gene product is a mitochondrial protein. Deletion of CYC2 leads to accumulation of apocytochrome c in the cytoplasm. However, strains with deletions of this gene still import low levels of cytochrome c into mitochondria. The effects of cyc2 mutations are more pronounced in rho- strains than in rho+ strains, even though rho- strains that are CYC2+ contain normal levels of holocytochrome c. cyc2 mutations affect levels of iso-1-cytochrome c more than they do levels of iso-2-cytochrome c, apparently because of the greater susceptibility of apo-iso-1-cytochrome c to degradation in the cytoplasm. We propose that CYC2 encodes a factor that increases the efficiency of cytochrome c import into mitochondria. PMID- 8413244 TI - PU.1 is a component of a multiprotein complex which binds an essential site in the murine immunoglobulin lambda 2-4 enhancer. AB - B-cell-specific enhancers have been identified in the immunoglobulin lambda locus 3' of each constant-region cluster. These enhancers contain two distinct domains, lambda A and lambda B, which are essential for enhancer function. lambda B contains a near-consensus binding site for the Ets family of transcription factors. In this study, we have identified a B-cell-specific protein complex which binds the lambda B motif of the lambda 2-4 enhancer in vitro and appears necessary for the activity of the enhancer in vivo, since mutations in lambda B which prevent this interaction also eliminate enhancer function. This complex contains PU.1, a member of the Ets family, and a transcriptional activator whose expression is restricted to cells of the hematopoietic system with the exception of T lymphocytes. In addition, it contains a factor which binds specifically to a region adjacent to the PU.1 binding site. This factor cannot bind lambda B autonomously but appears to require interaction with the PU.1 protein to stabilize its association with the DNA. This complex may be identical or related to the PU.1/NF-EM5 complex which interacts with a homologous DNA element in the immunoglobulin kappa 3' enhancer. PMID- 8413245 TI - The conserved C-terminal domain of the bovine papillomavirus E5 oncoprotein can associate with an alpha-adaptin-like molecule: a possible link between growth factor receptors and viral transformation. AB - The bovine papillomavirus E5 gene encodes an oncoprotein that can independently transform rodent fibroblasts. This small 44-amino-acid protein is thought to function through the activation of growth factor receptors. E5 activation of the epidermal growth factor receptor results in an increase in the number of activated receptors at the cell surface. This finding suggests that E5 may act by inhibiting the normal down regulation of activated epidermal growth factor receptor via coated pit-mediated endocytosis. We have constructed a fusion protein consisting of glutathione S-transferase and the conserved C-terminal domain of E5 (GST-E5) in order to identify E5-associated cellular proteins that may be involved in its transforming activity. We have identified a 125-kDa cellular protein with a strong associated serine kinase activity that specifically associated with GST-E5 in the reduced form but not with GST-E5 fusions that contained changes in several conserved amino acids. Microsequence and biochemical analyses suggest that p125 is a novel member of the alpha-adaptin family. Since alpha-adaptins have previously been shown to be involved in coated pit-mediated cell surface receptor endocytosis and down regulation, these results suggest that p125 may be an alpha-adaptin-like molecule involved in growth factor receptor down regulation and that E5 may act by inhibiting its activity. PMID- 8413246 TI - The MEF-3 motif is required for MEF-2-mediated skeletal muscle-specific induction of the rat aldolase A gene. AB - The rat aldolase A gene contains two alternative promoters and two alternative first exons. The distal promoter M is expressed at a high level only in skeletal muscle. Previous in vitro transfection studies identified the region from -202 to -85 as an enhancer that is responsible for dramatic activation during the differentiation of chicken primary myoblasts. This enhancer contains an A/T-rich sequence resembling the MEF-2 motif, which is an important element of muscle enhancers and promoters. In this study, we demonstrate that the MEF-2 sequence is essential but not sufficient for the activity of the enhancer. Another region required for the activity was recognized by a nuclear factor, tentatively named MAF1. MAF1 was found in both muscle cells and nonmuscle cells, and MAF1 from both cell types was indistinguishable by gel retardation and DNase I footprint experiments. The sequence required for MAF1 binding is very similar to the MEF-3 motif, which is an element of the skeletal muscle-specific enhancer of the cardiac troponin C gene. Because MAF1 and MEF-3 are closely related in both recognition sequence and distribution, MAF1 and MEF-3 probably represent the same nuclear factor which may play an important role in muscle gene transcription. Thus, the muscle-specific induction of the aldolase A gene is governed by muscle specific MEF-2 and existing MEF-3 (MAF1). PMID- 8413247 TI - XrpFI, an amphibian transcription factor composed of multiple polypeptides immunologically related to the GA-binding protein alpha and beta subunits, is differentially expressed during Xenopus laevis development. AB - XrpFI, first identified in the extract of Xenopus laevis oocyte nuclei, binds to a proximal sequence of the L14 ribosomal protein gene promoter. Its target sequence, 5'-TAACCGGAAGTTTGT-3', is required to fully activate the promoter, and the two G's of the central motif are essential for factor binding and transcriptional activation; our data also suggest that XrpFI may play a role in cap site positioning. The binding site of XrpFI is homologous to the sequence recognized by the family of ets genes. Antibodies specific for Ets-1 and Ets-2 proteins did not react with XrpFI, but those raised against the rat alpha and beta GA-binding proteins both supershifted the retarded bands formed by XrpFI. The Xenopus polypeptides related to GA-binding protein alpha interact with DNA both as monomers and as heterodimers associated with beta-related proteins. Oocyte nuclei contain multiple forms of alpha- and beta-related proteins: the alpha-like proteins remain throughout development, while the pattern of the beta species changes in the embryonic stages examined. beta-like proteins are undetectable in the cleavage period up to the neurula stage, but at later stages, when ribosomal protein genes are actively transcribed, two beta-related polypeptides reappear. PMID- 8413248 TI - A cis element required for induction of the interleukin 2 enhancer by human T cell leukemia virus type I binds a novel Tax-inducible nuclear protein. AB - The 40-kDa nuclear protein Tax encoded by human T-cell leukemia virus type I (HTLV-I) can transcriptionally activate the interleukin 2 (IL-2) enhancer even in the presence of the immunosuppressant cyclosporin A, which inhibits the activation of the IL-2 enhancer by T-cell mitogens. We have identified a Tax responsive element (TxRE) from -164 to -145 bp in the IL-2 enhancer which is sufficient to confer Tax responsiveness. A 45-kDa nuclear protein (TxRE-binding factor [TxREF]), present in Tax-expressing Jurkat cell lines but not in Jurkat cells without Tax, specifically interacts with the 5' TxRE sequence from -164 to 154. Deletion or mutation of this 5' TxRE sequence removes the binding of TxREF in vitro and dramatically reduces Tax activity in vivo. In addition, this site is responsible for the cyclosporin A-resistant expression of the IL-2 enhancer in the presence of Tax. Although the TxREF binding site contains an NF-kappa B like motif, UV cross-linking studies as well as gel retardation analysis reveal that TxREF is distinct from NF-kappa B. These results demonstrate that TxREF is a novel Tax-inducible DNA-binding protein and that TxRE plays a crucial role in mediating Tax-induced IL-2 gene expression. PMID- 8413249 TI - Inhibition of E2F-1 transactivation by direct binding of the retinoblastoma protein. AB - Loss of a functional retinoblastoma tumor suppressor gene product, pRB, is a key step in the development of many human tumors. pRB is a negative regulator of cell proliferation and appears to participate in control of entry into the S phase of the cell cycle. The recent demonstration that pRB binds to transcription factor E2F has provided a model for the mechanism of pRB-mediated growth regulation. Since adenovirus E1A proteins dissociate the pRB-E2F complexes and stimulate E2F dependent transcription, it has been suggested that pRB inhibits E2F transactivation. Although some evidence for this hypothesis has been provided, it has not been possible to determine the mechanism of pRB-mediated inhibition of E2F transactivation. In this study, we constructed mutants of E2F-1 that do not bind to pRB yet retain the ability to transactivate the adenovirus E2 promoter through E2F DNA-binding sites. We demonstrated that transactivation mediated by the wild-type E2F-1 protein was inhibited by overexpression of wild-type pRB but not by a naturally occurring mutant of pRB. Transactivation mediated by mutants of E2F-1 which do not bind to pRB was not affected by overexpression of wild-type pRB. Furthermore, when the E2F-1 transactivation domain was fused to the GAL4 DNA binding domain, pRB inhibited GAL4-E2F-1 transactivation through GAL4 sites. Expression of pRB did not inhibit transactivation mediated by GAL4-E2F-1 mutant constructs which were devoid of pRB binding. In conclusion, these data demonstrate that pRB inhibits E2F-dependent transactivation by direct protein protein interaction. PMID- 8413250 TI - DNA bending by retinoid X receptor-containing retinoid and thyroid hormone receptor complexes. AB - Retinoid X receptors (RXR) have been identified as common subunits in the regulation of multiple hormonal signaling pathways. Using circular permutation and phasing analysis of specific response elements, we present evidence that RXR retinoic acid receptor and RXR-thyroid hormone receptor heterodimer or RXR-RXR homodimer complexes induce directed DNA bends when bound to their cognate response elements. The extent of DNA bending induced by the RXR alpha-containing complexes varied and depended on the structure of the DNA-binding sites and the RXR partners. The overall bending orientation for RXR-containing complexes is directed toward the major groove of the DNA helix at the center of hormone response elements. Our observation implicates DNA bending as a possible mechanism underlying transcriptional regulation of distinct retinoid and thyroid hormone responsive genes. PMID- 8413251 TI - Nonrandom localization of recombination events in human alpha satellite repeat unit variants: implications for higher-order structural characteristics within centromeric heterochromatin. AB - Tandemly repeated DNA families appear to undergo concerted evolution, such that repeat units within a species have a higher degree of sequence similarity than repeat units from even closely related species. While intraspecies homogenization of repeat units can be explained satisfactorily by repeated rounds of genetic exchange processes such as unequal crossing over and/or gene conversion, the parameters controlling these processes remain largely unknown. Alpha satellite DNA is a noncoding tandemly repeated DNA family found at the centromeres of all human and primate chromosomes. We have used sequence analysis to investigate the molecular basis of 13 variant alpha satellite repeat units, allowing comparison of multiple independent recombination events in closely related DNA sequences. The distribution of these events within the 171-bp monomer is nonrandom and clusters in a distinct 20- to 25-bp region, suggesting possible effects of primary sequence and/or chromatin structure. The position of these recombination events may be associated with the location within the higher-order repeat unit of the binding site for the centromere-specific protein CENP-B. These studies have implications for the molecular nature of genetic recombination, mechanisms of concerted evolution, and higher-order structure of centromeric heterochromatin. PMID- 8413252 TI - Association of the human papillomavirus type 16 E7 protein with the S-phase specific E2F-cyclin A complex. AB - The transcription factor E2F has been shown to be involved in the expression of several cell cycle-regulated genes, and the activity of this factor is controlled by cellular proteins such as pRB and p107. E2F is also a target of the DNA virus oncoproteins (adenovirus E1A, simian virus 40 T antigen, and human papillomavirus [HPV] E7) (see the review by J. R. Nevins [Science 258: 424-429, 1992]). These viral oncoproteins dissociate an inactive complex between E2F and the retinoblastoma tumor suppressor protein (pRB), and this dissociation of the E2F pRB complex correlates with a stimulation of the E2F-dependent transcription. In the S phase of the cell cycle, E2F forms a complex with p107, cyclin A, and the cdk2 kinase (E2F-cyclin A complex). The cellular function of this S-phase specific complex is unclear. The adenovirus E1A protein dissociates the E2F cyclin A complex. The HPV type 16 (HPV-16) E7 protein, which possesses significant sequence homology with E1A, does not dissociate the E2F-cyclin A complex. We find that the HPV-16 E7 protein associates very efficiently with the E2F-cyclin A complex. This association is dependent on the sequences that are also necessary for the transforming activity of E7. Moreover, the E7 protein of a low-risk HPV (type 6b) is much less efficient in binding to the E2F-cyclin A complex compared with that of the high-risk type. We also find that the E2F cyclin A complex remains endogenously associated with the E7 protein in extracts of Caski cells, which express high levels of HPV-16 E7 protein. Finally, we have extensively purified the E2F-cyclin A complex from mouse L-cell extracts and show that, in cell extracts, the E2F-cyclin A complex remains associated with other cellular proteins. PMID- 8413253 TI - Binding of a phosphoprotein to the 3' untranslated region of the mouse protamine 2 mRNA temporally represses its translation. AB - The synthesis of the protamines, the predominant nuclear proteins of mammalian spermatozoa, is regulated during germ cell development by mRNA storage for about 7 days in the cytoplasm of differentiating spermatids. Two highly conserved sequences, the Y and H elements present in the 3' untranslated regions (UTRs) of all known mammalian protamine mRNAs, form RNA-protein complexes and specifically bind a protein of 18 kDa. Here, we show that translation of fusion mRNAs was markedly repressed in reticulocyte lysates supplemented with a mouse testis extract enriched for the 18-kDa protein when the mRNAs contained the 3' UTR of mouse protamine 2 (mP2) or the Y and H elements of mP2. No significant decrease was seen when the fusion mRNAs contained the 3' UTR of human growth hormone. The 18-kDa protein is developmentally regulated in male germ cells, requires phosphorylation for RNA binding, and is found in the ribonucleoprotein particle fractions of a testicular postmitochondrial supernatant. We propose that a phosphorylated 18-kDa protein plays a primary role in repressing translation of mP2 mRNA by interaction with the highly conserved Y and H elements. At a later stage of male gamete differentiation, the 18-kDa protein no longer binds to the mRNA, likely as a result of dephosphorylation, enabling the protamine mRNA to be translated. PMID- 8413254 TI - A family of human phosphodiesterases homologous to the dunce learning and memory gene product of Drosophila melanogaster are potential targets for antidepressant drugs. AB - We have isolated cDNAs for four human genes (DPDE1 through DPDE4) closely related to the dnc learning and memory locus of Drosophila melanogaster. The deduced amino acid sequences of the Drosophila and human proteins have considerable homology, extending beyond the putative catalytic region to include two novel, highly conserved, upstream conserved regions (UCR1 and UCR2). The upstream conserved regions are located in the amino-terminal regions of the proteins and appear to be unique to these genes. Polymerase chain reaction analysis suggested that these genes encoded the only homologs of dnc in the human genome. Three of the four genes were expressed in Saccharomyces cerevisiae and shown to encode cyclic AMP-specific phosphodiesterases. The products of the expressed genes displayed the pattern of sensitivity to inhibitors expected for members of the type IV, cyclic AMP-specific class of phosphodiesterases. Each of the four genes demonstrated a distinctive pattern of expression in RNA from human cell lines. PMID- 8413255 TI - Sequence-specific DNA primer effects on telomerase polymerization activity. AB - The ribonucleoprotein enzyme telomerase synthesizes one strand of telomeric DNA by copying a template sequence within the RNA moiety of the enzyme. Kinetic studies of this polymerization reaction were used to analyze the mechanism and properties of the telomerase from Tetrahymena thermophila. This enzyme synthesizes TTGGGG repeats, the telomeric DNA sequence of this species, by elongating a DNA primer whose 3' end base pairs with the template-forming domain of the RNA. The enzyme was found to act nonprocessively with short (10- to 12 nucleotide) primers but to become processive as TTGGGG repeats were added. Variation of the 5' sequences of short primers with a common 3' end identified sequence-specific effects which are distinct from those involving base pairing of the 3' end of the primer with the RNA template and which can markedly induce enzyme activity by increasing the catalytic rate of the telomerase polymerization reaction. These results identify an additional mechanistic basis for telomere and DNA end recognition by telomerase in vivo. PMID- 8413256 TI - Initiation and termination of DNA replication in human rRNA genes. AB - We have used the multicopy human rRNA genes as a model system to study replication initiation and termination in mammalian chromosomes. Enrichment for replicating molecules was achieved by isolating S-phase enriched populations of cells by centrifugal elutriation, purification of DNA associated with the nuclear matrix, and a chromatographic procedure that enriches for molecules containing single-stranded regions, a characteristic of replication forks. Two-dimensional agarose gel electrophoresis techniques were used to demonstrate that replication appears to initiate at multiple sites throughout most of the 31-kb nontranscribed spacer (NTS) of human ribosomal DNA but not within the 13-kb transcription unit or adjacent regulatory elements. Although initiation events were detected throughout the majority of the NTS, some regions may initiate more frequently than others. Termination of replication, the convergence of opposing replication forks, was found throughout the ribosomal DNA repeat units, and, in some repeats, specifically at the junction of the 3' end of the transcription unit and the NTS. This site-specific termination of replication is the result of pausing of replication forks near the sites of transcription termination. The naturally occurring multicopy rRNA gene family offers a unique system to study mammalian DNA replication without the use of chemical synchronization agents. PMID- 8413257 TI - Reconstitution of the Raf-1-MEK-ERK signal transduction pathway in vitro. AB - Raf-1 is a serine/threonine kinase which is essential in cell growth and differentiation. Tyrosine kinase oncogenes and receptors and p21ras can activate Raf-1, and recent studies have suggested that Raf-1 functions upstream of MEK (MAP/ERK kinase), which phosphorylates and activates ERK. To determine whether or not Raf-1 directly activates MEK, we developed an in vitro assay with purified recombinant proteins. Epitope-tagged versions of Raf-1 and MEK and kinase inactive mutants of each protein were expressed in Sf9 cells, and ERK1 was purified as a glutathione S-transferase fusion protein from bacteria. Raf-1 purified from Sf9 cells which had been coinfected with v-src or v-ras was able to phosphorylate kinase-active and kinase-inactive MEK. A kinase-inactive version of Raf-1 purified from cells that had been coinfected with v-src or v-ras was not able to phosphorylate MEK. Raf-1 phosphorylation of MEK activated it, as judged by its ability to stimulate the phosphorylation of myelin basic protein by glutathione S-transferase-ERK1. We conclude that MEK is a direct substrate of Raf 1 and that the activation of MEK by Raf-1 is due to phosphorylation by Raf-1, which is sufficient for MEK activation. We also tested the ability of protein kinase C to activate Raf-1 and found that, although protein kinase C phosphorylation of Raf-1 was able to stimulate its autokinase activity, it did not stimulate its ability to phosphorylate MEK. PMID- 8413258 TI - The intracisternal A-particle upstream element interacts with transcription factor YY1 to activate transcription: pleiotropic effects of YY1 on distinct DNA promoter elements. AB - Murine intracisternal A-particle long terminal repeats contain an intracisternal A-particle upstream enhancer (IUE) element that binds to a 65-kDa IUE binding protein (IUEB) present in both undifferentiated F9 embryonal carcinoma cells and differentiated parietal endoderm-like PYS-2 cells. This IUE element confers a CpG methylation-sensitive IUEB binding and enhancer activity. Using gel retardation, methylation interference, CpG methylation sensitivity binding, and cotransfection assays, we have now identified the 65-kDa IUEB as YY1 (also called NF-E1, delta, or UCRBP), a zinc finger protein related to the Kruppel family. YY1 binds to a number of similar but distinct DNA motifs, and cotransfection assays indicate that these motifs have different enhancer potentials in PYS-2 cells. The relative strengths of these elements are as follows: IUE > kappa E3' from the human immunoglobulin kappa light-chain 3' enhancer > upstream conserved region from the Moloney murine leukemia virus promoter. Results of DNA binding assays suggest that the differences in enhancer potentials are due to the different binding affinities of YY1 to the various motifs and the binding of two other transcription factors to the IUE sequence. PMID- 8413259 TI - A regulatory element in the beta 2-microglobulin promoter identified by in vivo footprinting. AB - Expression of the beta 2-microglobulin (beta 2-m) and major histocompatibility complex (MHC) class I genes is coordinately regulated. By ligation-mediated polymerase chain reaction, we have analyzed in vivo factor binding to the promoter region of the murine beta 2-m gene. In adult spleen, in which beta 2-m is expressed, strong protection was found in three elements. Two of these elements, the beta 2-m NF-kappa B binding site and the interferon consensus sequence, are homologous to the regulatory elements of the MHC class I genes and were also found to be protected in spleen. A third protected element, PAM, identified in this work, is unique to the beta 2-m gene. None of the elements showed protection in brain tissue, in which neither the beta 2-m nor the MHC class I gene is expressed. In vivo footprinting was also performed with F9 embryonal carcinoma cells, in which expression of the beta 2-m and MHC class I genes is induced at a low level only upon stimulation with retinoic acid (RA). No in vivo protection was detected before and after RA treatment of F9 cells, indicating that RA induction of beta 2-m (and MHC class I) expression occurs without detectable in vivo factor occupancy, whereas EL4 T lymphocytes expressing beta 2-m at a high level exhibited strong protection similar to that in spleen. Despite the lack of in vivo occupancy, the nuclear factors specific for each of the three elements were present in brain tissue and F9 cells as well as in spleen tissue and EL4 cells. We show that PAM, an element identified by its in vivo protection, binds nuclear factors ranging from 40 to 50 kDa in size and is capable of enhancing transcription of a reporter in F9 and other cells. Taken together, these results indicate that in vivo factor occupancy for the beta 2-m and MHC class I promoters is coordinated and occurs through a mechanism other than mere expression of relevant factors. PMID- 8413260 TI - Overproduction of Rb protein after the G1/S boundary causes G2 arrest. AB - The Rb protein is known to exert its activity at decision points in the G1 phase of the cell cycle. To investigate whether it may also play some role(s) at later points in the cell cycle, we used a system of rapid inducible gene amplification to conditionally overexpress Rb protein during G2 phase. A cell line expressing a temperature-sensitive simian virus 40 large T antigen (T-Ag) was stably transfected with plasmids containing the Rb cDNA linked to the simian virus 40 origin of replication: pRB-wt, pRB-fs, and pRB-Dra, carrying wild-type murine Rb cDNA, a frameshift mutation close to the beginning of the Rb coding region, and a single-amino-acid deletion in the E1A/T-Ag binding pocket, respectively. Numerous independent cell lines were isolated at the nonpermissive temperature; cell lines displaying a high level of episomal amplification of an intact Rb expression cassette following shiftdown to the permissive temperature were chosen for further analysis. Plasmid pRB-fs did not express detectable Rb antigen, while pRB Dra expressed full-length Rb protein. The Dra mutation has previously been shown to abrogate phosphorylation as well as T-Ag binding. Fluorescence-activated cell sorting (FACS) analysis revealed that cultures induced to overexpress either wild type or Dra mutant Rb proteins were significantly enriched for cells with a G2 DNA content. Cultures that amplified pRB-fs or rearranged pRB-wt and did not express Rb protein had normal cell cycle profiles. Double-label FACS analysis showed that cells overexpressing Rb or Rb-Dra proteins were uniformly accumulating in G2, whereas cells expressing endogenous levels of Rb were found throughout the cell cycle. These results indicate that Rb protein is interacting with some component(s) of the cell cycle-regulatory machinery during G2 phase. PMID- 8413261 TI - Insulin-stimulated oocyte maturation requires insulin receptor substrate 1 and interaction with the SH2 domains of phosphatidylinositol 3-kinase. AB - Xenopus oocytes from unprimed frogs possess insulin-like growth factor I (IGF-I) receptors but lack insulin and IGF-I receptor substrate 1 (IRS-1), the endogenous substrate of this kinase, and fail to show downstream responses to hormonal stimulation. Microinjection of recombinant IRS-1 protein enhances insulin stimulated phosphatidylinositol (PtdIns) 3-kinase activity and restores the germinal vesicle breakdown response. Activation of PtdIns 3-kinase results from formation of a complex between phosphorylated IRS-1 and the p85 subunit of PtdIns 3-kinase. Microinjection of a phosphonopeptide containing a pYMXM motif with high affinity for the src homology 2 (SH2) domain of PtdIns 3-kinase p85 inhibits IRS 1 association with and activation of the PtdIns 3-kinase. Formation of the IRS-1 PtdIns 3-kinase complex and insulin-stimulated PtdIns 3-kinase activation are also inhibited by microinjection of a glutathione S-transferase fusion protein containing the SH2 domain of p85. This effect occurs in a concentration-dependent fashion and results in a parallel loss of hormone-stimulated oocyte maturation. These inhibitory effects are specific and are not mimicked by glutathione S transferase fusion proteins expressing the SH2 domains of ras-GAP or phospholipase C gamma. Moreover, injection of the SH2 domains of p85, ras-GAP, and phospholipase C gamma do not interfere with progesterone-induced oocyte maturation. These data demonstrate that phosphorylation of IRS-1 plays an essential role in IGF-I and insulin signaling in oocyte maturation and that this effect occurs through interactions of the phosphorylated YMXM/YXXM motifs of IRS 1 with SH2 domains of PtdIns 3-kinase or some related molecules. PMID- 8413262 TI - Phosphatidylinositol 3-kinase activity is important for progesterone-induced Xenopus oocyte maturation. AB - In somatic cells, phosphatidylinositol 3-kinase (PI3 kinase) is a critical intermediary in growth factor-induced mitogenesis. We have examined the role of this enzyme in meiotic maturation of Xenopus laevis oocytes. PI3 kinase activity was present in immunoprecipitates of the p85 subunit of PI3 kinase from immature oocytes and markedly increased following progesterone stimulation. Injection of bacterially expressed protein corresponding to the C-terminal SH2 domain of p85 (SH2-C) inhibited progesterone-induced PI3 kinase activation and meiotic maturation. Injection of protein corresponding to the N-terminal SH2 domain or the SH3 domain of p85 did not inhibit PI3 kinase activation or maturation. SH2-C did not inhibit oocyte maturation induced by c-mos RNA injection. In addition, radiolabelled SH2-C was used to probe oocyte lysates, revealing that a novel 200 kDa protein bound to SH2-C. This protein may be an important mediator of progesterone-induced lipid metabolism in oocytes. PMID- 8413263 TI - Delayed chromosomal instability induced by DNA damage. AB - DNA damage induced by ionizing radiation can result in gene mutation, gene amplification, chromosome rearrangements, cellular transformation, and cell death. Although many of these changes may be induced directly by the radiation, there is accumulating evidence for delayed genomic instability following X-ray exposure. We have investigated this phenomenon by studying delayed chromosomal instability in a hamster-human hybrid cell line by means of fluorescence in situ hybridization. We examined populations of metaphase cells several generations after expanding single-cell colonies that had survived 5 or 10 Gy of X rays. Delayed chromosomal instability, manifested as multiple rearrangements of human chromosome 4 in a background of hamster chromosomes, was observed in 29% of colonies surviving 5 Gy and in 62% of colonies surviving 10 Gy. A correlation of delayed chromosomal instability with delayed reproductive cell death, manifested as reduced plating efficiency in surviving clones, suggests a role for chromosome rearrangements in cytotoxicity. There were small differences in chromosome destabilization and plating efficiencies between cells irradiated with 5 or 10 Gy of X rays after a previous exposure to 10 Gy and cells irradiated only once. Cell clones showing delayed chromosomal instability had normal frequencies of sister chromatid exchange formation, indicating that at this cytogenetic endpoint the chromosomal instability was not apparent. The types of chromosomal rearrangements observed suggest that chromosome fusion, followed by bridge breakage and refusion, contributes to the observed delayed chromosomal instability. PMID- 8413264 TI - A CRE/ATF-like site in the upstream regulatory sequence of the human interleukin 1 beta gene is necessary for induction in U937 and THP-1 monocytic cell lines. AB - Transfection of U937 and THP-1 cells with a recombinant plasmid, pIL1(4.0kb)-CAT, containing 4 kb of the interleukin 1 beta (IL-1 beta) gene upstream regulatory sequence resulted in inducer-dependent expression of chloramphenicol acetyltransferase activity. Treatment of the transfected cells with various combinations of the inducers lipopolysaccharide, phorbol myristate acetate, and dibutyryl cyclic AMP upregulated the IL-1 beta promoter. In U937 and THP-1 cells, maximum stimulation of both the endogenous IL-1 beta gene and pIL1(4.0kb)-CAT transfectants was observed following treatment with the combination of inducing agents lipopolysaccharide-phorbol myristate acetate-dibutyryl cyclic AMP. This combination of inducing agents was used to identify and study, at the molecular level, some of the regulatory elements necessary for induction of the IL-1 beta gene. A series of 5' deletion derivatives of the upstream regulatory sequence were used in transient transfection assays to identify an 80-bp fragment located between -2720 and -2800 bp upstream of the mRNA start site that was required for induction. Exonuclease III mapping, electrophoretic mobility shift assays (EMSA), and DNA sequence analysis of this region were used to identify a transcription factor binding sequence which contained a potential cyclic AMP response element (CRE/ATF)- and NF-kappa B-like binding site. Site-directed mutagenesis of the CRE/ATF-like site resulted in the loss of binding of a specific factor or factors as determined by EMSA. The loss of binding activity directly correlated with a loss of approximately 75% of promoter activity as determined in transient transfection assays. As determined by EMSA, the factor binding to the CRE/ATF like site was present in nuclear extracts prepared from both uninduced and induced THP-1 and U937 cells. However, the intensity of the band appeared to be increased when nuclear extracts from induced cells were used. In contrast to the CRE/ATF mutation, which resulted in the loss of promoter activity, mutation of the NF-kappa B-like site resulted in a moderate increase in activity in U937 cells. A similar increase in promoter activity was not observed in THP-1 cells. From these studies, we conclude that a CRE/ATF-like site and a factor or factors interacting with this site are essential for the maximum induction of the IL-1 beta gene in stimulated U937 and THP-1 cells. PMID- 8413265 TI - Identification of a killer cell-specific regulatory element of the mouse perforin gene: an Ets-binding site-homologous motif that interacts with Ets-related proteins. AB - The gene encoding the cytolytic protein perforin is selectively expressed by activated killer lymphocytes. To understand the mechanisms underlying the cell type-specific expression of this gene, we have characterized the regulatory functions and the DNA-protein interactions of the 5'-flanking region of the mouse perforin gene (Pfp). A region extending from residues +62 through -141, which possesses the essential promoter activity, and regions further upstream, which are able to either enhance or suppress gene expression, were identified. The region between residues -411 and -566 was chosen for further characterization, since it contains an enhancer-like activity. We have identified a 32-mer sequence (residues -491 to -522) which appeared to be capable of enhancing gene expression in a killer cell-specific manner. Within this segment, a 9-mer motif (5' ACAGGAAGT-3', residues -505 to -497; designated NF-P motif), which is highly homologous to the Ets proto-oncoprotein-binding site, was found to interact with two proteins, NF-P1 and NF-P2. NF-P2 appears to be induced by reagents known to up-regulate the perforin message level and is present exclusively in killer cells. Electrophoretic mobility shift assay and UV cross-linking experiments revealed that NF-P1 and NF-P2 may possess common DNA-binding subunits. However, the larger native molecular mass of NF-P1 suggests that NF-P1 contains an additional non-DNA-binding subunit(s). In view of the homology between the NF-P motif and other Ets proto-oncoprotein-binding sites, it is postulated that NF-P1 and NF-P2 belong to the Ets protein family. Results obtained from the binding competition assay, nevertheless, suggest that NF-P1 and NF-P2 are related to but distinct from Ets proteins, e.g., Ets-1, Ets-2, and NF-AT/Elf-1, known to be expressed in T cells. PMID- 8413266 TI - DNA topoisomerase I controls the kinetics of promoter activation and DNA topology in Saccharomyces cerevisiae. AB - Inactivation of the nonessential TOP1 gene, which codes for Saccharomyces cerevisiae DNA topoisomerase I, affects the rate of transcription starting at the ADH2 promoter. For both the chromosomal gene and the plasmid-borne promoter, mRNA accumulation is kinetically favored in the mutant relative to a wild-type isogenic strain. The addition of ethanol causes in wild-type yeast strains a substantial increase in linking number both on the ADH2-containing plasmid and on the resident 2 microns DNA. Evidence has been obtained that such an in vivo increase in linking number depends on (i) the activity of DNA topoisomerase I and of no other enzyme and (ii) ethanol addition, not on the release from glucose repression. A direct cause-effect relationship between the change in supercoiling and alteration of transcription cannot be defined. However, the hypothesis that a metabolism-induced modification of DNA topology in a eukaryotic cell plays a role in regulating gene expression is discussed. PMID- 8413267 TI - Dimerization mediated through a leucine zipper activates the oncogenic potential of the met receptor tyrosine kinase. AB - Oncogenic activation of the met (hepatocyte growth factor/scatter factor) receptor tyrosine kinase involves a genomic rearrangement that generates a hybrid protein containing tpr-encoded sequences at its amino terminus fused directly to the met-encoded receptor kinase domain. Deletion of Tpr sequences abolishes the transforming ability of this protein, implicating this region in oncogenic activation. We demonstrate, by site-directed mutagenesis and coimmunoprecipitation experiments, that a leucine zipper motif within Tpr mediates dimerization of the tpr-met product and is essential for the transforming activity of the met oncogene. By analogy with ligand-stimulated activation of receptor tyrosine kinases, we propose that constitutive dimerization mediated by a leucine zipper motif within Tpr is responsible for oncogenic activation of the Met kinase. The possibility that this mechanism of activation represents a paradigm for a class of receptor tyrosine kinase oncogenes activated by DNA rearrangement is discussed. PMID- 8413268 TI - Function of the growth-regulated transcription initiation factor TIF-IA in initiation complex formation at the murine ribosomal gene promoter. AB - Alterations in the rate of cell proliferation are accompanied by changes in the transcription of rRNA genes. In mammals, this growth-dependent regulation of transcription of genes coding for rRNA (rDNA) is due to reduction of the amount or activity of an essential transcription factor, called TIF-IA. Extracts prepared from quiescent cells lack this factor activity and, therefore, are transcriptionally inactive. We have purified TIF-IA from exponentially growing cells and have shown that it is a polypeptide with a molecular mass of 75 kDa which exists as a monomer in solution. Using a reconstituted transcription system consisting of purified transcription factors, we demonstrate that TIF-IA is a bona fide transcription initiation factor which interacts with RNA polymerase I. Preinitiation complexes can be assembled in the absence of TIF-IA, but formation of the first phosphodiester bonds of nascent rRNA is precluded. After initiation, TIF-IA is liberated from the initiation complex and facilitates transcription from templates bearing preinitiation complexes which lack TIF-IA. Despite the pronounced species specificity of class I gene transcription, this growth dependent factor has been identified not only in mouse but also in human cells. Murine TIF-IA complements extracts from both growth-inhibited mouse and human cells. The analogous human activity appears to be similar or identical to that of TIF-IA. Therefore, despite the fact that the RNA polymerase transcription system has evolved sufficiently rapidly that an rDNA promoter from one species will not function in another species, the basic mechanisms that adapt ribosome synthesis to cell proliferation have been conserved. PMID- 8413269 TI - Functional interaction of the v-Rel and c-Rel oncoproteins with the TATA-binding protein and association with transcription factor IIB. AB - Rel family proteins regulate the expression of genes linked to kappa B-binding motifs. Little is known, however, of the mechanism by which they enhance transcription. We have investigated the ability of the v-Rel and c-Rel oncoproteins to interact with components of the basal transcription machinery. Here we report that both the acidic transcription activation domain mapping to the unique C terminus of chicken c-Rel and the F9 cell-specific activation region common to both v-Rel and c-Rel interact with the TATA-binding protein (TBP) and transcription factor IIB (TFIIB) in vitro and in vivo. We also demonstrate that TPB interaction with Rel activation regions leads to synergistic activation of transcription of a kappa B-linked reporter gene. Combined with the observation that the mouse c-Rel and human RelA proteins also interact with TBP and TFIIB in vitro, these results suggest that association with basal transcription factors is important for the transcriptional activities of Rel family proteins. PMID- 8413270 TI - Regulation of the human cardiac/slow-twitch troponin C gene by multiple, cooperative, cell-type-specific, and MyoD-responsive elements. AB - The cardiac troponin C (cTnC) gene produces identical transcripts in slow-twitch skeletal muscle and in heart muscle (R. Gahlmann, R. Wade, P. Gunning, and L. Kedes, J. Mol. Biol. 201:379-391, 1988). A separate gene encodes the fast-twitch skeletal muscle troponin C and is not expressed in heart muscle. We have used transient transfection to characterize the regulatory elements responsible for skeletal and cardiac cell-type-specific expression of the human cTnC (HcTnC) gene. At least four separate elements cooperate to confer tissue-specific expression of this gene in differentiated myotubes; a basal promoter (between -61 and -13) augments transcription 9-fold, upstream major regulatory sequences (between -68 and -142 and between -1319 and -4500) augment transcription as much as 39-fold, and at least two enhancer-like elements in the first intron (between +58 and +1028 and between +1029 and +1523) independently augment transcription 4- to 5-fold. These enhancers in the first intron increase myotube-specific chloramphenicol acetyltransferase activity when linked to their own promoter elements or to the heterologous simian virus 40 promoter, and the effects are multiplicative rather than additive. Each of the major myotube regulatory regions is capable of responding directly or indirectly to the myogenic determination factor, MyoD.A MyoD expression vector in 10T1/2 cells induced constructs carrying either the upstream HcTnC promoter elements or the first intron of the gene 300- to 500-fold. Expression was inhibited by cotransfection with Id, a negative regulator of basic helix-loop-helix transcription factors. The basal promoter contains five tandem TGGGC repeats that interact with Sp1 or an Sp1-like factor in nuclear extracts. Mutational analysis of this element demonstrated that two of the five repeat sequences were sufficient to support basal level muscle cell specific transcription. Whereas the basal promoter is also critical for expression in cardiac myocytes, the elements upstream of -67 appear to play little or no role. Major augmentation of expression in cardiomyocytes is also provided by sequences in the first intron, but these are upstream (between +58 and +1028). The downstream segment of the first intron has no enhancer activity in cardiomyocytes. A specific DNA-protein complex is formed by this C2 cell enhancer with extracts from C2 cells but not cardiomyocytes. These observations suggest that tissue-specific expression of the HcTnC gene is cooperatively regulated by the complex interactions of multiple regulatory elements and that different elements are used to regulate expression in myogenic and cardiac cells. PMID- 8413271 TI - Molecular analysis of the differential hepatic expression of rat kininogen family genes. AB - Serum concentration of rat T1 kininogen increases 20- to 30-fold in response to acute inflammation, an induced hepatic synthesis regulated primarily at the transcriptional level. We have demonstrated by transient transfection analyses that rat T1 kininogen gene/chloramphenicol acetyltransferase (T1K/CAT) constructs are highly responsive to interleukin-6 and dexamethasone. In these studies we examined the regulation of a highly homologous K kininogen gene promoter and showed that it is minimally induced under identical conditions. The basal expression of the KK/CAT construct was, however, five- to sevenfold higher than that of the analogous T1K/CAT construct. Promoter-swapping experiments to examine the molecular basis of this differentially regulated basal expression showed that at least two K kininogen promoter regions are important for conferring its high basal expression: a distal 19-bp region (C box) constituted a binding site for C/EBP family proteins, and a proximal 66-bp region contained two adjacent binding sites for hepatocyte nuclear factor 3 (HNF-3). While the C box in the K kininogen promoter was able to interact with C/EBP transcription factors, the T1 kininogen promoter C box could not. In addition, HNF-3 binding sites of the K kininogen promoter demonstrated stronger affinities than those of the T1 kininogen promoter. Since C/EBP and HNF-3 are highly enriched in the liver and are known to enhance transcription of liver-specific genes, these differences in their binding activities thus accounted for the K kininogen gene's higher basal expression. Our studies demonstrated that evolutionary divergence of a few critical nucleotides may lead to subtle changes in the binding affinities of a transcription factor to its recognition site, profoundly altering expression of the downstream gene. PMID- 8413272 TI - Hyperactive recombination in the mitochondrial DNA of the natural death nuclear mutant of Neurospora crassa. AB - In Neurospora crassa, a recessive mutant allele of a nuclear gene, nd (natural death), causes rapid degeneration of the mitochondrial DNA, a process that is manifested phenotypically as an accelerated form of senescence in growing and stationary mycelia. To examine the mechanisms that are involved in the degradation of the mitochondrial chromosome, several mitochondrial DNA restriction fragments unique to the natural-death mutant were cloned and characterized through restriction, hybridization, and nucleotide sequence analyses. All of the cloned DNA pieces contained one to four rearrangements that were generated by unequal crossing-over between direct repeats of several different nucleotide sequences that occur in pairs and are dispersed throughout the mitochondrial chromosome of wild-type Neurospora strains. The most abundant repeats, a family of GC-rich sequences that includes the so-called PstI palindromes, were not involved in the generation of deletions in the nd mutant. The implication of these results is that the nd allele hyperactivates a general system for homologous recombination in the mitochondria of N. crassa. Therefore, the nd+ allele either codes for a component of the complex of proteins that catalyzes recombination, and possibly repair and replication, of the mitochondrial chromosome or specifies a regulatory factor that controls the synthesis or activity of at least one enzyme or ancillary factor that is affiliated with mitochondrial DNA metabolism. PMID- 8413273 TI - The HRIGRXXR region of the DEAD box RNA helicase eukaryotic translation initiation factor 4A is required for RNA binding and ATP hydrolysis. AB - eIF-4A is a eukaryotic translation initiation factor that is required for mRNA binding to ribosomes. It exhibits single-stranded RNA-dependent ATPase activity, and in combination with a second initiation factor, eIF-4B, it exhibits duplex RNA helicase activity. eIF-4A is the prototype of a large family of proteins termed the DEAD box protein family, whose members share nine highly conserved amino acid regions. The functions of several of these conserved regions in eIF-4A have previously been assigned to ATP binding, ATPase, and helicase activities. To define the RNA-binding region of eIF-4A, a UV-induced cross-linking assay was used to analyze binding of mutant eIF-4A proteins to RNA. Mutants carrying mutations in the ATP-binding region (AXXXXGKT), ATPase region (DEAD), helicase region (SAT), and the most carboxy-terminal conserved region of the DEAD family, HRIGRXXR, were tested for RNA cross-linking. We show that mutations, either conservative or not, in any one of the three arginines in the HRIGRXXR sequence drastically reduced eIF-4A cross-linking to RNA. In addition, all the mutations in the HRIGRXXR region abrogate RNA helicase activity. Some but not all of these mutations affect ATP binding and ATPase activity. This is consistent with the hypothesis that the HRIGRXXR region is involved in the ATP hydrolysis reaction and would explain the coupling of ATPase and RNA-binding/helicase activities. Our results show that the HRIGRXXR region, which is QRXGRXXR or QXXGRXXR in the RNA and DNA helicases of the helicase superfamily II, is involved in ATP hydrolysis dependent RNA interaction during unwinding. We also show that mutations in other regions of eIF-4A that abolish ATPase activity sharply decrease eIF-4A cross linking to RNA. A model is proposed in which eIF-4A first binds ATP, resulting in a change in eIF-4A conformation which allows RNA binding that is dependent on the HRIGRXXR region. Binding of RNA induces ATP hydrolysis, leading to a more stable interaction with RNA. This process is then linked to unwinding of duplex RNA in the presence of eIF-4B. PMID- 8413274 TI - The major histocompatibility complex class II promoter-binding protein RFX (NF-X) is a methylated DNA-binding protein. AB - A mammalian protein called RFX or NF-X binds to the X box (or X1 box) in the promoters of a number of major histocompatibility (MHC) class II genes. In this study, RFX was shown to have the same DNA-binding specificity as methylated DNA binding protein (MDBP), and its own cDNA was found to contain a binding site for MDBP in the leader region. MDBP is a ubiquitous mammalian protein that binds to certain DNA sequences preferentially when they are CpG methylated and to other related sequences, like the X box, irrespective of DNA methylation. MDBP from HeLa and Raji cells formed DNA-protein complexes with X-box oligonucleotides that coelectrophoresed with those containing standard MDBP sites. Furthermore, MDBP and X-box oligonucleotides cross-competed for the formation of these DNA-protein complexes. DNA-protein complexes obtained with MDBP sites displayed the same partial supershifting with an antiserum directed to the N terminus of RFX seen for complexes containing an X-box oligonucleotide. Also, the in vitro-transcribed translated product of a recombinant RFX cDNA bound specifically to MDBP ligands and displayed the DNA methylation-dependent binding of MDBP. RFX therefore contains MDBP activity and thereby also EF-C, EP, and MIF activities that are indistinguishable from MDBP and that bind to methylation-independent sites in the transcriptional enhancers of polyomavirus and hepatitis B virus and to an intron of c-myc. PMID- 8413275 TI - Structural requirements of 5S rRNA for nuclear transport, 7S ribonucleoprotein particle assembly, and 60S ribosomal subunit assembly in Xenopus oocytes. AB - Structural requirements of 5S rRNA for nuclear transport and RNA-protein interactions have been studied by analyzing the behavior of oocyte-type 5S rRNA and of 31 different in vitro-generated mutant transcripts after microinjection into the cytoplasm of Xenopus oocytes. Experiments reveal that the sequence and secondary and/or tertiary structure requirements of 5S rRNA for nuclear transport, storage in the cytoplasm as 7S ribonucleoprotein particles, and assembly into 60S ribosomal subunits are complex and nonidentical. Elements of loops A, C, and E, helices II and V, and bulged and hinge nucleotides in the central domain of 5S rRNA carry the essential information for these functional activities. Assembly of microinjected 5S rRNA into 60S ribosomal subunits was shown to occur in the nucleus; thus, the first requirement for subunit assembly is nuclear targeting. The inhibitory effects of ATP depletion, wheat germ agglutinin, and chilling on the nuclear import of 5S rRNA indicate that it crosses the nuclear envelope through the nuclear pore complex by a pathway similar to that used by karyophilic proteins. PMID- 8413276 TI - Extensive editing of CR2 maxicircle transcripts of Trypanosoma brucei predicts a protein with homology to a subunit of NADH dehydrogenase. AB - Several genes of the Trypanosoma brucei mitochondrial genome (the maxicircle) encode mRNAs that are so extensively altered by RNA editing that the gene cannot be identified by analysis of the DNA sequence. The 322-nucleotide preedited RNA of one of these genes, CR2, is converted into a 647-nucleotide transcript by the addition of 345 uridines and the deletion of 20 genomically encoded uridines. The fully edited transcript has an open reading frame that predicts a 194-amino-acid protein. This protein, which we name ND9 (NADH dehydrogenase subunit 9), has homology to a subunit of NADH dehydrogenase (respiratory complex I). Seven guide RNAs that can specify edited CR2 sequence have been identified. Steady-state levels of unedited ND9 transcripts are greater in bloodstream than in procyclic forms, but edited ND9 mRNA is present in similar abundance in both life cycle stages. PMID- 8413277 TI - Short artificial hairpins sequester splicing signals and inhibit yeast pre-mRNA splicing. AB - To examine the stability of yeast (Saccharomyces cerevisiae) pre-mRNA structures, we inserted a series of small sequence elements that generated potential RNA hairpins at the 5' splice site and branch point regions. We analyzed spliceosome assembly and splicing in vitro as well as splicing and nuclear pre-mRNA retention in vivo. Surprisingly, the inhibition of in vivo splicing approximately paralleled that of in vitro splicing. Even a 6-nucleotide hairpin could be shown to inhibit splicing, and a 15-nucleotide hairpin gave rise to almost complete inhibition. The in vitro results indicate that hairpins that sequester the 5' splice site have a major effect on the early steps of spliceosome assembly, including U1 small nuclear ribonucleoprotein binding. The in vivo experiments lead to comparable conclusions as the sequestering hairpins apparently result in the transport of pre-mRNA to the cytoplasm. The observations are compared with previous data from both yeast and mammalian systems and suggest an important effect of pre-mRNA structure on in vivo splicing. PMID- 8413278 TI - The tumor suppressor p53 and the oncoprotein simian virus 40 T antigen bind to overlapping domains on the MDM2 protein. AB - The oncogene mdm2 has been found to be amplified in human sarcomas, and the gene product binds to the tumor suppressor p53. In this report, we describe the dissection of the MDM2-binding domain on p53 as well as the p53-binding domain on MDM2. We also demonstrate that the oncoprotein simian virus 40 T antigen binds to the product of cellular oncogene mdm2. We have constructed several N- and C terminal deletion mutants of p53 and MDM2, expressed them in vitro, and assayed their in vitro association capability. The N-terminal boundary of the p53-binding domain on MDM2 is between amino acids 1 and 58, while the C-terminal boundary is between amino acids 221 and 155. T antigen binds to an overlapping domain on the MDM2 protein. On the other hand, the MDM2-binding domain of p53 is defined by amino acids 1 and 159 at the N terminus. At the C terminus, binding is progressively reduced as amino acids 327 to 145 are deleted. We determined the effect of human MDM2 on the transactivation ability of wild-type human p53 in the Saos-2 osteosarcoma cell line, which does not have any endogenous p53. Human MDM2 inhibited the ability of human p53 to transactivate the promoter with p53-binding sites. Thus, human MDM2 protein, like the murine protein, can inactivate the transactivation ability of human p53. Interestingly, both the transactivation domain and the MDM2-binding domain of p53 are situated near the N terminus. We further show that deletion of the N-terminal 58 amino acids of MDM2, which eliminates p53 binding, also abolishes the capability of inactivating p53 mediated transactivation. This finding suggests a correlation of in vitro p53 MDM2 binding with MDM2's ability in vivo to interfere with p53-mediated transactivation. PMID- 8413279 TI - Transcriptional activity of the zinc finger protein NGFI-A is influenced by its interaction with a cellular factor. AB - NGFI-A is an immediate-early gene that encodes a transcription factor whose DNA binding domain is composed of three zinc fingers. To define the domains responsible for its transcriptional activity, a mutational analysis was conducted with an NGFI-A molecule in which the zinc fingers were replaced by the GAL4 DNA binding domain. In a cotransfection assay, four activation domains were found within NGFI-A. Three of the activation domains are similar to those characterized previously: one contains a large number of acidic residues, another is enriched in proline and glutamine residues, and another has some sequence homology to a domain found in Krox-20. The fourth bears no resemblance to previously described activation domains. NGFI-A also contains an inhibitory domain whose removal resulted in a 15-fold increase in NGFI-A activity. This increase in activity occurred in all mammalian cell types tested but not in Drosophila S2 cells. Competition experiments in which increasing amounts of the inhibitory domain were cotransfected along with NGFI-A demonstrated a dose-dependent increase in NGFI-A activity. A point mutation within the inhibitory domain of the competitor (I293F) abolished this property. When the analogous mutation was introduced into native NGFI-A, a 17-fold increase in activity was observed. The inhibitory effect therefore appears to be the result of an interaction between this domain and a titratable cellular factor which is weakened by this mutation. Downmodulation of transcription factor activity through interaction with a cellular factor has been observed in several other systems, including the regulation of transcription factor E2F by retinoblastoma protein, and in studies of c-Jun. PMID- 8413280 TI - Transcription of alpha-specific genes in Saccharomyces cerevisiae: DNA sequence requirements for activity of the coregulator alpha 1. AB - Transcription activation of alpha-specific genes in Saccharomyces cerevisiae is regulated by two proteins, MCM1 and alpha 1, which bind to DNA sequences, called P'Q elements, found upstream of alpha-specific genes. Neither MCM1 nor alpha 1 alone binds efficiently to P'Q elements. Together, however, they bind cooperatively in a manner that requires both the P' sequence, which is a weak binding site for MCM1, and the Q sequence, which has been postulated to be the binding site for alpha 1. We analyzed a collection of point mutations in the P'Q element of the STE3 gene to determine the importance of individual base pairs for alpha-specific gene transcription. Within the 10-bp conserved Q sequence, mutations at only three positions strongly affected transcription activation in vivo. These same mutations did not affect the weak binding to P'Q displayed by MCM1 alone. In vitro DNA binding assays showed a direct correlation between the ability of the mutant sequences to form ternary P'Q-MCM1-alpha 1 complexes and the degree to which transcription was activated in vivo. Thus, the ability of alpha 1 and MCM1 to bind cooperatively to P'Q elements is critical for activation of alpha-specific genes. In all natural alpha-specific genes the Q sequence is adjacent to the degenerate side of P'. To test the significance of this geometry, we created several novel juxtapositions of P, P', and Q sequences. When the Q sequence was opposite the degenerate side, the composite QP' element was inactive as a promoter element in vivo and unable to form stable ternary QP'-MCM1-alpha 1 complexes in vitro. We also found that addition of a Q sequence to a strong MCM1 binding site allows the addition of alpha 1 to the complex. This finding, together with the observation that Q-element point mutations affected ternary complex formation but not the weak binding of MCM1 alone, supports the idea that the Q sequence serves as a binding site for alpha 1. PMID- 8413281 TI - AFR1 acts in conjunction with the alpha-factor receptor to promote morphogenesis and adaptation. AB - Mating pheromone receptors activate a G-protein signaling pathway that induces changes in transcription, cell division, and morphogenesis needed for the conjunction of Saccharomyces cerevisiae. The C terminus of the alpha-factor pheromone receptor functions in two complex processes, adaptation and morphogenesis. Adaptation to alpha-factor may occur through receptor desensitization, and alpha-factor-induced morphogenesis forms the conjugation bridge between mating cells. A plasmid overexpression strategy was used to isolate a new gene, AFR1, which acts together with the receptor C terminus to promote adaptation. The expression of AFR1 was highly induced by alpha-factor. Unexpectedly, cells lacking AFR1 showed a defect in alpha-factor-stimulated morphogenesis that was similar to the morphogenesis defect observed in cells producing C-terminally truncated alpha-factor receptors. In contrast, AFR1 overexpression resulted in longer projections of morphogenesis, which suggests that this gene may directly stimulate morphogenesis. These results indicate that AFR1 encodes a developmentally regulated function that coordinates both the regulation of receptor signaling and the induction of morphogenesis during conjugation. PMID- 8413282 TI - Homologous and illegitimate recombination in developing Xenopus oocytes and eggs. AB - Exogenous DNA is efficiently recombined when injected into the nuclei of Xenopus laevis oocytes. This reaction proceeds by a homologous resection-annealing mechanism which depends on the activity of a 5'-->3' exonuclease. Two possible functions for this recombination activity have been proposed: it may be a remnant of an early process in oogenesis, such as meiotic recombination or amplification of genes coding for rRNA, or it may reflect materials stored for embryogenesis. To test these hypotheses, recombination capabilities were examined with oocytes at various developmental stages. Late-stage oocytes performed only homologous recombination, whereas the smallest oocytes ligated the restriction ends of the injected DNA but supported no homologous recombination. This transition from ligation to recombination activity was also seen in nuclear extracts from these same stages. Exonuclease activity was measured in the nuclear extracts and found to be low in early stages and then to increase in parallel with recombination capacity in later stages. The accumulation of exonuclease and recombination activities during oogenesis suggests that they are stored for embryogenesis and are not present for oocyte-specific functions. Eggs were also tested and found to catalyze homologous recombination, ligation, and illegitimate recombination. Retention of homologous recombination in eggs is consistent with an embryonic function for the resection-annealing mechanism. The observation of all three reactions in eggs suggests that multiple pathways are available for the repair of double-strand breaks during the extremely rapid cleavage stages after fertilization. PMID- 8413283 TI - Transcriptional control of the chicken cardiac myosin light-chain gene is mediated by two AT-rich cis-acting DNA elements and binding of serum response factor. AB - Transcriptional control of the cardiac/slow skeletal alkali myosin light-chain (MLC1c/1s) gene is mediated, in part, by two highly conserved AT-rich cis-acting elements present in the immediate 5' flanking region. These elements cooperate to form an enhancer that can impart tissue specificity to heterologous promoters that are themselves not tissue specific in their pattern of expression. In the chicken, one of these elements matches the binding site for myocyte-specific enhancer-binding factor 2, while the other is a cis-acting element present in the transcriptional control regions of all striated alkali MLC genes (except MLC3f) and is referred to as the MLC box. The central decanucleotide core region of the MLC box closely resembles the CArG (CC[A/T]6GG) box of the serum response element, and the binding of muscle nuclear protein complexes to this element can be competed for with a synthetic serum response element. On the basis of their competition profiles and requirements for nonspecific competitor, two nuclear protein complexes, which compete for binding to the CArG-like region of the MLC box, have been identified. One of the complexes binds to a mutation of the CArG like region that inactivates transcription of a linked reporter gene, while binding of the other complex is inhibited by this mutation. This latter complex reacts with an antibody to serum response factor (SRF) and exhibits the same binding characteristics as purified SRF. These results demonstrate that transcriptional control of the chicken MLC1c/1s gene resides in an upstream enhancer that is composed of two separate AT-rich elements, both of which are required to drive expression of a linked reporter gene. The binding of a nuclear protein complex containing SRF to one of these elements, the MLC box, is required for gene activation and apparently inhibited by other nuclear factors whose binding overlaps that of the SRF complex. PMID- 8413284 TI - Identification of C/EBP basic region residues involved in DNA sequence recognition and half-site spacing preference. AB - C/EBP and GCN4 are basic region-leucine zipper (bZIP) DNA-binding proteins that recognize the dyad-symmetric sequences ATTGCGCAAT and ATGAGTCAT, respectively. The sequence specificities of these and other bZIP proteins are determined by their alpha-helical basic regions, which are related at the primary sequence level. To identify amino acids that are responsible for the different DNA sequence specificities of C/EBP and GCN4, two kinds of hybrid proteins were constructed: GCN4-C/EBP chimeras fused at various positions in the basic region and substitution mutants in which GCN4 basic region amino acids were replaced by the corresponding residues from C/EBP. On the basis of the DNA-binding characteristics of these hybrid proteins, three residues that contribute significantly to the differences in C/EBP and GCN4 binding specificity were defined. These residues are clustered along one face of the basic region alpha helix. Two of these specificity residues were not identified as DNA-contacting amino acids in a recently reported crystal structure of a GCN4-DNA complex, suggesting that the residues used by C/EBP and GCN4 to make base contacts are not identical. A random binding site selection procedure also was used to define the optimal recognition sequences for three of the GCN4-C/EBP fusion proteins. These experiments identify an element spanning the hinge region between the basic region and leucine zipper domains that dictates optimal half-site spacing (either directly abutted for C/EBP or overlapping by one base pair for GCN4) in high affinity binding sites for these two proteins. PMID- 8413285 TI - Ribosomal pausing during translation of an RNA pseudoknot. AB - The genomic RNA of the coronavirus infectious bronchitis virus contains an efficient ribosomal frameshift signal which comprises a heptanucleotide slippery sequence followed by an RNA pseudoknot structure. The presence of the pseudoknot is essential for high-efficiency frameshifting, and it has been suggested that its function may be to slow or stall the ribosome in the vicinity of the slippery sequence. To test this possibility, we have studied translational elongation in vitro on mRNAs engineered to contain a well-defined pseudoknot-forming sequence. Insertion of the pseudoknot at a specific location within the influenza virus PB1 mRNA resulted in the production of a new translational intermediate corresponding to the size expected for ribosomal arrest at the pseudoknot. The appearance of this protein was transient, indicating that it was a true paused intermediate rather than a dead-end product, and mutational analysis confirmed that its appearance was dependent on the presence of a pseudoknot structure within the mRNA. These observations raise the possibility that a pause is required for the frameshift process. The extent of pausing at the pseudoknot was compared with that observed at a sequence designed to form a simple stem-loop structure with the same base pairs as the pseudoknot. This structure proved to be a less effective barrier to the elongating ribosome than the pseudoknot and in addition was unable to direct efficient ribosomal frameshifting, as would be expected if pausing plays an important role in frameshifting. However, the stem-loop was still able to induce significant pausing, and so this effect alone may be insufficient to account for the contribution of the pseudoknot to frameshifting. PMID- 8413286 TI - V(D)J recombination coding junction formation without DNA homology: processing of coding termini. AB - Coding junction formation in V(D)J recombination generates diversity in the antigen recognition structures of immunoglobulin and T-cell receptor molecules by combining processes of deletion of terminal coding sequences and addition of nucleotides prior to joining. We have examined the role of coding end DNA composition in junction formation with plasmid substrates containing defined homopolymers flanking the recombination signal sequence elements. We found that coding junctions formed efficiently with or without terminal DNA homology. The extent of junctional deletion was conserved independent of coding ends with increased, partial, or no DNA homology. Interestingly, G/C homopolymer coding ends showed reduced deletion regardless of DNA homology. Therefore, DNA homology cannot be the primary determinant that stabilizes coding end structures for processing and joining. PMID- 8413287 TI - Activation of the beta-globin promoter by the locus control region correlates with binding of a novel factor to the CAAT box in murine erythroleukemia cells but not in K562 cells. AB - Four distinct factors in extracts from murine erythroleukemia (MEL) cells interacted with the human beta-globin gene promoter CAAT box: CP1, GATA-1, and two novel factors, denoted a and b, one of which is highly inducible in the MEL system. GATA-1 binding to the CAAT element was very unstable (half-life < 1 min), whereas bindings of a, b, and CP1 were comparatively stable, with half-lives of 18, 19, and 3.5 min, respectively. Stable transfections of MEL cells showed that in the presence of the beta-globin locus control region (LCR), the wild-type CAAT box, a mutant which bound to GATA-1 with increased stability over the normal sequences, and a mutant which bound a, b, and CP1 specifically could all stimulate transcription greater than ninefold over that induced by a null CAAT mutation in both uninduced and terminally differentiated MEL cells. A mutant which bound the a and b factors specifically gave only a twofold stimulation of promoter activity, and this lower activity correlated with a decrease in the stability of binding of the b protein. On the other hand, CP1 binding alone did not stimulate transcription. Taken together, these results suggest that in the context of the wild-type beta-globin CAAT element the b factor stimulates transcription directed by the LCR in MEL cells, although the LCR can also function through more stable GATA-1-binding sequences. However, in K562 cells, the wild-type beta-globin CAAT box alone was unable to stimulate gene expression directed by the LCR and high levels of transcription were obtained only upon inclusion of more upstream beta-globin promoter sequences. In contrast, a construct containing only the A gamma-globin CAAT box region did give high expression levels in K562 cells. Thus, there is a fundamental difference in the way the LCR functions in these two model systems in terms of its requirements at the promoter level. PMID- 8413288 TI - A portion of RNA polymerase II molecules has a component essential for stress responses and stress survival. AB - Cells respond to stress by altering gene expression, and these adjustments facilitate stress tolerance. Although transcriptional changes are integral to most stress responses, little is known about the mechanisms that permit the transcription apparatus itself to tolerate stress. Here we report that a major role of the RNA polymerase II subunit RPB4 is to permit appropriate transcriptional responses during stress. Yeast cells lacking RPB4 have essentially wild-type growth rates at moderate temperatures (18 to 22 degrees C), but their growth rates are substantially reduced at temperatures outside this range. When subjected to a heat shock, cells lacking RPB4 rapidly lose the ability to transcribe genes and experience a dramatic loss in viability. When cells lacking RPB4 are subjected to the nutrient stress that accompanies entry into stationary phase, they also exhibit a substantial decline in mRNA synthesis and in viability relative to wild-type cells. Interestingly, the portion of RNA polymerase II molecules that contain RPB4 is small in log phase but increases substantially as cells enter stationary phase. We propose that the association of RPB4 with the other RNA polymerase II subunits increases the tolerance of the enzyme to stress. PMID- 8413289 TI - UVB-induced DNA breaks interfere with transcriptional induction of c-fos. AB - Oxidative stress may play an important role in the carcinogenic action of UVB light (290 to 320 nm). UVB light induces the growth-related immediate-early gene c-fos in JB6 mouse epidermal cells, but at the same time it causes structural damage to DNA, in particular DNA strand breakage. We have studied the effect of the modulation of the frequencies of DNA breaks on the transcriptional induction of c-fos by changing the cellular antioxidant defense or by inhibiting break repair. Reduction of UVB-induced DNA breakage in a stable transfectant with an increased complement of glutathione peroxidase enhanced the induction of c-fos. In contrast, c-fos induction was diminished in stable transfectants with Cu,Zn superoxide dismutase. Increasing the stationary concentration of UVB-induced DNA breaks by inhibition of repair in the presence of the adenosine diphosphoribose (ADPR)-transferase inhibitor 3-amino-benzamide suppressed the induction of c-fos. We conclude that DNA breaks which are induced by UVB via oxidative processes interfere with the transcriptional induction of c-fos. DNA breaks appear to exert a long-range effect on chromatin conformation which is incompatible with efficient transcription. This notion is supported by the observation that inhibition of break rejoining by 3-amino-benzamide suppressed the UVB induction of the endogenous c-fos gene and of a stably integrated construct containing the c-fos regulatory sequences linked to a reporter gene. In contrast, the induction of the same construct was not inhibited when it remained extrachromosomal in transient transfection experiments. PMID- 8413290 TI - P-element-induced interallelic gene conversion of insertions and deletions in Drosophila melanogaster. AB - We studied the process by which whd, a P-element insertion allele of the Drosophila melanogaster white locus, is replaced by its homolog in the presence of transposase. These events are interpreted as the result of double-strand gap repair following excision of the P transposon in whd. We used a series of alleles derived from whd through P-element mobility as templates for this repair. One group of alleles, referred to collectively as whd-F, carried fragments of the P element that had lost some of the sequences needed in cis for mobility. The other group, whd-D, had lost all of the P insert and had some of the flanking DNA from white deleted. The average replacement frequencies were 43% for whd-F alleles and 7% for the whd-D alleles. Some of the former were converted at frequencies exceeding 50%. Our data suggest that the high conversion frequencies for the whd F templates can be attributed at least in part to an elevated efficiency of repair of unexpanded gaps that is possibly caused by the closer match between whd F sequences and the unexpanded gap endpoints. In addition, we found that the gene substitutions were almost exclusively in the direction of whd being replaced by the whd-F or whd-D allele rather than the reverse. The template alleles were usually unaltered in the process. This asymmetry implies that the conversion process is unidirectional and that the P fragments are not good substrates for P element transposase. Our results help elucidate a highly efficient double-strand gap repair mechanism in D. melanogaster that can also be used for gene replacement procedures involving insertions and deletions. They also help explain the rapid spread of P elements in populations. PMID- 8413291 TI - cis-acting sequences of the rat troponin I slow gene confer tissue- and development-specific transcription in cultured muscle cells as well as fiber type specificity in transgenic mice. AB - Transcription of the genes coding for troponin I slow (TnIslow) and other contractile proteins is activated during skeletal muscle differentiation, and their expression is later restricted to specific fiber types during maturation. We have isolated and characterized the rat TnIslow gene in order to begin elucidating its regulation during myogenesis. Transcriptional regulatory regions were delineated by using constructs, containing TnIslow gene sequences driving the expression of the chloramphenicol acetyltransferase (CAT) reporter gene, that were transiently transfected into undifferentiated and differentiated C2C12 cells. TnIslow 5'-flanking sequences directed transcription specifically in differentiated cells. However, transcription rates were approximately 10-fold higher in myotubes transfected with constructs containing the 5'-flanking sequences plus the intragenic region residing upstream of the translation initiation site (introns 1 and 2), indicative of interactions between elements residing upstream and in the introns of the gene. Deletion analysis of the 5' region of the TnIslow gene showed that the 200 bp upstream of the transcription initiation site is sufficient to confer differentiation-specific transcription in C2C12 myocytes. MyoD consensus binding sites were found both in the upstream 200 bp region and in a region residing in the second intron that is highly homologous to the quail TnIfast enhancer. Transactivation experiments using transfected NIH 3T3 fibroblasts with TnI-CAT constructs containing intragenic and/or upstream sequences and with the myogenic factors MyoD, myogenin, and MRF4 showed different potentials of these factors to induce transcription. Transgenic mice harboring the rat TnI-CAT fusion gene expressed the reporter specifically in the skeletal muscle. Furthermore, CAT levels were approximately 50-fold higher in the soleus than in the extensor digitorum longus, gastrocnemius, or tibialis muscle, indicating that the regulatory elements that restrict TnI transcription to slow twitch myofibers reside in the sequences we have analyzed. PMID- 8413292 TI - Identification of distinct roles for separate E1A domains in disruption of E2F complexes. AB - The adenovirus E1A protein can disrupt protein complexes containing the E2F transcription factor in association with cellular regulatory proteins such as the retinoblastoma gene product (Rb) and the Rb-related p107 protein. Previous experiments have shown that the CR1 and CR2 domains of E1A are required for this activity. We now demonstrate that the CR2 domain is essential for allowing E1A to interact with the E2F-Rb or the E2F-p107-cyclin A-cdk2 complex. Multimeric complexes containing E1A can be detected when the CR1 domain has been rendered inactive by mutation. In addition, the E1A CR1 domain, but not the CR2 domain, is sufficient to prevent the interaction of E2F with Rb or p107. On the basis of these results, we suggest a model whereby the CR2 domain brings E1A to the E2F complexes and then, upon a normal equilibrium dissociation of Rb or p107 from E2F, the E1A CR1 domain is able to block the site of interaction on Rb or p107, thereby preventing the re-formation of the complexes. PMID- 8413293 TI - Expression of a retroposon-like sequence upstream of the putative Trypanosoma brucei variant surface glycoprotein gene expression site promoter. AB - We have cloned the region spanning the putative promoter from two variant surface glycoprotein gene expression sites that are at each end of chromosome M4 of Trypanosoma brucei IsTat 7. Both expression sites contain a retroposon-like sequence (ESR) pseudogene whose 3' end is approximately 30 bp upstream of the putative expression site promoter. The ESRs from both expression sites share considerable sequence homology and are related to LINE-like elements, especially the T. brucei ingi retroposon. Other ESRs are located on large, but not intermediate or mini-, chromosomes in the IsTaR 1 serodeme, and the total copy number is 10 to 20, similar to that estimated for variant surface glycoprotein expression sites. No DNA rearrangements in the vicinity of the ESR and putative expression site promoter were detected following antigenic switches in the IsTaR 1 serodeme. ESR transcripts are present in bloodstream, but not procyclic, forms. Variation in transcript size and sequence between bloodstream variant antigenic types implies that only the ESR from the active expression site is transcribed. This pattern of expression reflects that of sequences downstream of the putative expression site promoter, suggesting that the region of coordinately controlled expression extends upstream of this promoter. PMID- 8413294 TI - The ZEBRA activation domain: modular organization and mechanism of action. AB - An RNA polymerase II activator often contains several regions that contribute to its potency, an organization ostensibly analogous to the modular architecture of promoters and enhancers. The regulatory significance of this parallel organization has not been systematically explored. We considered this problem by examining the activation domain of the Epstein-Barr virus transactivator ZEBRA. We performed our experiments in vitro so that the activator concentrations, stabilities, and affinities for DNA could be monitored. ZEBRA and various amino terminal deletion derivatives, expressed in and purified from Escherichia coli, were assayed in a HeLa cell nuclear extract for the ability to activate model reporter templates bearing one, three, five, and seven upstream ZEBRA binding sites. Our data show that ZEBRA contains four modules that contribute to its potency in vitro. The modules operate interchangeably with promoter sites to determine the transcriptional response such that the loss of modules can be compensated for by increasing promoter sites. Potassium permanganate footprinting was used to show that transcriptional stimulation is a consequence of the activator's ability to promote preinitiation complex assembly. Kinetic measurements of transcription complex assembly in a reconstituted system indicate that ZEBRA promotes formation of a subcomplex requiring the TFIIA and TFIID fractions, where TFIIA acts as an antirepressor. We propose a model in which the concentration of DNA-bound activation modules in the vicinity of the gene initiates synergistic transcription complex assembly. PMID- 8413295 TI - Identification and characterization of an Alu-containing, T-cell-specific enhancer located in the last intron of the human CD8 alpha gene. AB - Expression of the human CD8 alpha gene is restricted to cells of the lymphoid lineage and developmentally regulated during thymopoiesis. As an initial step towards understanding the molecular basis for tissue-specific expression of this gene, we surveyed the surrounding chromatin structure for potential cis-acting regulatory regions by DNase I hypersensitivity mapping and found four hypersensitive sites, three of which were T cell restricted. By using a reporter based expression approach, a T-cell-specific enhancer was identified by its close association with a prominent T-cell-restricted hypersensitive sites in the last intron of the CD8 alpha gene. Deletion studies demonstrated that the minimal enhancer is adjacent to a negative regulatory element. DNA sequence analysis of the minimal enhancer revealed a striking cluster of consensus binding sites for Ets-1, TCF-1, CRE, GATA-3, LyF-1, and bHLH proteins which were verified by electrophoretic mobility shift assays. In addition, the 5' end of the enhancer was composed of an Alu repeat which contained the GATA-3, bHLH, and LyF-1 binding sites. Site-directed mutation of the Ets-1 and GATA-3 sites dramatically reduced enhancer activity. The functional importance of the other binding sites only became apparent when combinations of mutations were analyzed. Taken together, these results suggest that the human CD8 alpha gene is regulated by the interaction of multiple T-cell nuclear proteins with a transcriptional enhancer located in the last intron of the gene. Comparison of the CD8 alpha enhancer with other recently identified T-cell-specific regulatory elements suggests that a common set of transcription factors regulates several T-cell genes. PMID- 8413296 TI - Biological and biochemical activities of a chimeric epidermal growth factor-Elk receptor tyrosine kinase. AB - Eph, Elk, and Eck are prototypes of a large family of transmembrane protein tyrosine kinases, which are characterized by a highly conserved cysteine-rich domain and two fibronectin type III repeats in their extracellular regions. Despite the extent of the Eph family, no extracellular ligands for any family member have been identified, and hence, little is known about the biological and biochemical properties of these receptor-like tyrosine kinases. In the absence of a physiological ligand for the Elk receptor, we constructed chimeric receptor molecules, in which the extracellular region of the Elk receptor is replaced by the extracellular, ligand-binding domain of the epidermal growth factor (EGF) receptor. These chimeric receptors were expressed in NIH 3T3 cells that lack endogenous EGF receptors to analyze their signaling properties. The chimeric EGF Elk receptors became glycosylated, were correctly localized to the plasma membrane, and bound EGF with high affinity. The chimeric receptors underwent autophosphorylation and induced the tyrosine phosphorylation of a specific set of cellular proteins in response to EGF. EGF stimulation also induced DNA synthesis in fibroblasts stably expressing the EGF-Elk receptors. In contrast, EGF stimulation of these cells did not lead to visible changes in cellular morphology, nor did it induce loss of contact inhibition in confluent monolayers or growth in semisolid media. The Elk cytoplasmic domain is therefore able to induce tyrosine phosphorylation and DNA synthesis in response to an extracellular ligand, suggesting that Elk and related polypeptides function as ligand-dependent receptor tyrosine kinases. PMID- 8413297 TI - Transcription of the rat glucagon gene by the cyclic AMP response element-binding protein CREB is modulated by adjacent CREB-associated proteins. AB - The cyclic AMP (cAMP) response element (CRE) of the rat glucagon gene (Glu-CRE, 5'-TGACGTCA-3') mediates transcriptional responses to 8-bromo-cAMP and protein kinase A (PKA) in a glucagon-producing hamster islet cell line (InR1G9). By several different DNA-protein binding assays, we show that the transcription factor CREB binds to the CRE octamer and that additional nuclear proteins bind to sequences adjacent to the CRE. Mutation of the Glu-CRE octamer attenuates both the binding of CREB and cAMP-dependent PKA-stimulated transcriptional activity in transient transfection experiments but does not affect the binding of adjacent CREB-associated proteins. Progressive deletions and clustered point mutations of the sequences flanking the Glu-CRE identify sequences (5'-TCATT-3') located both 5' and 3' to the core CRE octamer that bind several proteins. Two proteins with molecular masses of 80 and 100 kDa bind to each of the 5' and 3' TCATT sites. Formation of additional protein-DNA complexes containing 45- and 20-kDa proteins depends upon the integrity of both TCATT sequences. Deletion or point mutation of the TCATT motif located on the 3' side of the CRE octamer results in enhanced transcriptional responses to PKA, suggesting that the CREB-associated proteins decrease the ability of CREB to mediate PKA-stimulated transcription. Results from these studies demonstrate that nucleotides flanking the core CRE octamer can influence the activity of the CRE by serving as binding sites for proteins that modulate the function of CREB and suggest a mechanism to explain why some consensus palindromic CREs are less responsive to cAMP stimulation than others. PMID- 8413298 TI - urbs1, a gene regulating siderophore biosynthesis in Ustilago maydis, encodes a protein similar to the erythroid transcription factor GATA-1. AB - Ustilago maydis secretes ferrichrome-type siderophores, ferric-ion-binding compounds, in response to iron starvation. TA2701, a non-enterobactin-producing, non-ferrichrome-utilizing mutant of Salmonella typhimurium LT-2, was employed as a biological indicator in a novel screening method to isolate three N-methyl-N' nitro-N-nitrosoguanidine-induced U. maydis mutants defective in the regulation of ferrichrome-type siderophore biosynthesis. These mutants displayed a constitutive phenotype; they produced siderophores in the presence of iron concentrations that would typically repress siderophore synthesis in wild-type strains. A 4.8-kb fragment of U. maydis genomic DNA capable of restoring normal regulation of siderophore biosynthesis in the constitutive mutants was identified. This segment of DNA contains an intronless open reading frame that specifies a protein of 950 amino acids containing two finger motifs similar to those found in the erythroid transcription factor GATA-1. Disruption of this open reading frame in a wild-type strain gave rise to cells that produced siderophores constitutively. Genetic studies indicated that the disruption mutation was allelic to the chemically induced mutations, confirming that the structural gene for a regulator rather than a suppressor gene had been cloned. Northern (RNA) analysis of the gene revealed a 4.2-kb transcript that is expressed constitutively at low levels in wild-type cells. The data support the hypothesis that this gene, which we designate urbs1 (Ustilago regulator of biosynthesis of siderophores), acts directly or indirectly to repress biosynthesis of siderophores in U. maydis. PMID- 8413299 TI - Isolation and characterization of fifteen ecdysone-inducible Drosophila genes reveal unexpected complexities in ecdysone regulation. AB - Our insights into the regulatory mechanisms by which the steroid hormone ecdysone triggers Drosophila melanogaster metamorphosis have largely depended on puffs in the larval salivary gland polytene chromosomes as a means of identifying genes of interest. Here, we describe an approach that provides access to ecdysone inducible genes that are expressed in most larval and imaginal tissues, regardless of their ability to form puffs in the polytene chromosomes. Several hundred cDNAs were picked at random from subtracted cDNA libraries and subjected to a rapid and sensitive screen for their ability to detect mRNAs induced by ecdysone in the presence of cycloheximide. Of the 15 genes identified in this manner, 2 correspond to early puffs in the salivary gland polytene chromosomes, at 63F and 75B, confirming that this screen functions at the desired level of sensitivity and is capable of identifying novel primary-response genes. Three of the genes, Eig45-1, Eig58, and Eig87, are expressed coordinately with the salivary gland early genes; one of them, Eig58, maps to the 58BC puff that is active when the 74EF and 75B early puffs are at their maximal size. Another gene identified in this screen, Eig17-1, encodes a novel cytochrome P-450. On the basis of its sequence identity and temporal profile of expression, this gene may play a role in steroid hormone metabolism and thus could provide a mechanism for feedback regulation of ecdysone production. Although all 15 genes have patterns of transcription that are consistent with ecdysone regulation in vivo, 5 genes do not appear to be induced by the late larval ecdysone pulse. This indicates that ecdysone induction in larval organs cultured with cycloheximide is not always indicative of a primary response to the hormone. PMID- 8413300 TI - The SH2 domain is required for stable phosphorylation of p56lck at tyrosine 505, the negative regulatory site. AB - The catalytic function of Src-related tyrosine protein kinases is repressed by phosphorylation of a conserved carboxy-terminal tyrosine residue. Recent studies suggest that this inhibitory event is not the result of autophosphorylation but that it is mediated by another cytoplasmic tyrosine protein kinase, termed p50csk. In this report, we have evaluated the processes regulating the extent of phosphorylation of the inhibitory carboxy-terminal tyrosine residue of p56lck, a lymphocyte-specific member of the Src family. By analyzing kinase-defective variants of p56lck expressed in mouse NIH 3T3 cells, we have found that the noncatalytic Src homology 2 (SH2) domain, but not the SH3 sequence or the sites of Lck myristylation and autophosphorylation, is necessary for stable phosphorylation at the carboxy-terminal tyrosine 505. Further studies in which Lck and Csk were coexpressed in S. cerevisiae indicated that the absence of the SH2 domain did not affect the ability of Csk to phosphorylate p56lck at tyrosine 505. However, we observed that incubation of cells with the tyrosine phosphatase inhibitor pervanadate restored the tyrosine 505 phosphorylation of Lck polypeptides devoid of the SH2 motif. Additionally, the presence of the SH2 sequence protected tyrosine 505 from in vitro dephosphorylation by the hemopoietic tyrosine protein phosphatase CD45. Taken together, these findings raised the possibility that the SH2 motif contributes to the physiological suppression of the catalytic function of p56lck at least in part through its ability to stabilize phosphorylation at the inhibitory site. PMID- 8413301 TI - Biphasic ordered induction of heme synthesis in differentiating murine erythroleukemia cells: role of erythroid 5-aminolevulinate synthase. AB - During dimethyl sulfoxide (DMSO)-stimulated differentiation of murine erythroleukemia (MEL) cells, one of the early events is the induction of the heme biosynthetic pathway. While recent reports have clearly demonstrated that GATA-1 is involved in the induction of erythroid cell-specific forms of 5 aminolevulinate synthase (ALAS-2) and porphobilinogen (PBG) deaminase and that cellular iron status plays a regulatory role for ALAS-2, little is known about regulation of the remainder of the pathway. In the current study, we have made use of a stable MEL cell mutant (MEAN-1) in which ALAS-2 enzyme activity is not induced by DMSO, hexamethylene bisacetamide (HMBA), or butyric acid. In this cell line, addition of 2% DMSO to growing cultures results in the normal induction of PBG deaminase and coproporphyrinogen oxidase but not in the induction of the terminal two enzymes, protoporphyrinogen oxidase and ferrochelatase. These DMSO treated cells did not produce mRNA for beta-globin and do not terminally differentiate. In addition, the cellular level of ALAS activity declines rapidly after addition of DMSO, indicating that ALAS-1 must turn over rapidly at this time. Addition of 75 microM hemin alone to the cultures did not induce cells to terminally differentiate or induce any of the pathway enzymes. However, the simultaneous addition of 2% DMSO and 75 microM hemin caused the cells to carry out a normal program of terminal erythroid differentiation, including the induction of ferrochelatase and beta-globin. These data suggest that induction of the entire heme biosynthetic pathway is biphasic in nature and that induction of the terminal enzymes may be mediated by the end product of the pathway, heme. We have introduced mouse ALAS-2 cDNA into the ALAS-2 mutant cell line (MEAN-1) under the control of the mouse metallothionein promoter (MEAN-RA). When Cd and Zn are added to cultures of MEAN-RA in the absence of DMSO, ALAS-2 is induced but erythroid differentiation does not occur and cells continue to grow normally. In the presence of metallothionein inducers and DMSO, the MEAN-RA cells induce in a fashion similar to that found with the wild-type 270 MEL cells. Induction of the activities of ALAS, PBG deaminase, coproporphyrinogen oxidase, and ferrochelatase occurs. In cultures of MEAN-RA where ALAS-2 had been induced with Cd plus Zn 24 h prior to DMSO addition, onset of heme synthesis occurs more rapidly than when DMSO and Cd plus Zn are added simultaneously. This study reveals that induction of ALAS-2 alone is not sufficient to induce terminal differentiation of the MEAN RA cells, and it does not appear that ALAS-2 alone is the rate-limiting enzyme of the heme biosynthetic pathway during MEL cell differentiation. PMID- 8413302 TI - Two novel transmembrane protein tyrosine kinases expressed during Caenorhabditis elegans hypodermal development. AB - We describe our characterization of kin-15 and kin-16, a tandem pair of homologous Caenorhabditis elegans genes encoding transmembrane protein tyrosine kinases (PTKs) with an unusual structure: the predicted extracellular domain of each putative gene product is only about 50 amino acids, and there are no potential autophosphorylation sites in the C-terminal domain. Using lacZ fusions, we found that kin-15 and kin-16 both appear to be expressed during postembryonic development in the large hypodermal syncytium (hyp7) around the time that specific hypodermal cells fuse with hyp7. kin-15 and kin-16 were positioned on the genetic and physical maps, but extrachromosomal arrays containing wild-type kin-15 and/or kin-16 genes were unable to complement candidate lethal mutations. The results suggest that kin-15 and kin-16 may be specifically involved in cell cell interactions regulating cell fusions that generate the hypodermis during postembryonic development. PMID- 8413303 TI - The murine alpha B-crystallin/small heat shock protein enhancer: identification of alpha BE-1, alpha BE-2, alpha BE-3, and MRF control elements. AB - The murine alpha B-crystallin gene (a member of the small heat shock protein family) is expressed constitutively at high levels in the lens and at lower levels in many other tissues, including skeletal muscle. We have previously used the herpes simplex virus thymidine kinase promoter fused to the human growth hormone gene to identify an alpha B-crystallin enhancer at positions -427 to -259 that has high activity in muscle and low activity in lens cell lines. In the study reported here, we performed DNase I footprinting, transfection, mutagenesis, and electrophoretic mobility shift experiments using the murine C2C12 muscle and alpha TN4-1 lens cell lines and the rabbit N/N1003A lens cell line to identify sequences responsible for activity of this enhancer. Enhancer activity in both the muscle and lens cells was dependent on novel elements called alpha BE-1 (-407 to -397), alpha BE-2 (-360 to -327), and alpha BE-3 (-317 to 306). These elements were also weakly occupied by nuclear proteins in L929 cells, which appear to express the alpha B-crystallin gene at a very low level (detectable only by the polymerase chain reaction). A fourth element containing a consensus muscle regulatory factor-binding site called MRF (-300 to -288) was occupied and used only by the C2C12 muscle cells. Cotransfection in NIH 3T3 cells and antibody-gel shift experiments using C2C12 nuclear extracts indicated that MyoD, myogen, or a similar member of this family can activate the alpha B crystallin enhancer by interaction with the MRF site. Taken together, we conclude that the alpha BE-1, alpha BE-2, and alpha BE-3 elements are shared by both lens and muscle cells, but the MRF element is used only in muscle cells, providing the first example of a muscle-specific control element in a crystallin gene. PMID- 8413304 TI - MyoD and myogenin act on the chicken myosin light-chain 1 gene as distinct transcriptional factors. AB - Expression of MyoD, myogenin, MRF4, and Myf-5 converts nonmuscle cells to muscle cells. In an attempt to analyze the roles of these factors, we have investigated their effects on transcription driven by the promoter of the chicken myosin alkaline light-chain (MLC1) gene. The activation by CMD1 or c-myogenin (chicken MyoD or myogenin, respectively) was dependent on the existence of a muscle specific regulatory region located from positions -2096 to -1743. Its distal half, containing a pair of E boxes (CANNTG), had been previously characterized as an enhancer responsive to CMD1 but not to c-myogenin. In this study, we report the identification of another enhancer in the muscle-specific regulatory region which is preferentially responsive to c-myogenin. Deletion and mutation analyses indicated that this enhancer requires a single E box and its flanking sequences. Furthermore, analysis of chimeric proteins of CMD1 and c-myogenin indicated that regions outside the basic helix-loop-helix domain of c-myogenin are involved in the specificity of the enhancer. These results show that CMD1 and c-myogenin act on the MLC1 gene by recognizing different upstream DNA sequences and that direct or indirect interactions between the regions outside the basic helix-loop-helix domain and flanking sequences of E boxes are involved in the target sequence specificity. PMID- 8413305 TI - Ets proteins: new factors that regulate immunoglobulin heavy-chain gene expression. AB - We used a DNA-protein interaction screening method to isolate a cDNA, Erg-3, whose product binds to a site, designated pi, present in the immunoglobulin (Ig) heavy-chain gene enhancer. Erg-3 is an alternatively spliced product of the erg gene and contains an Ets DNA-binding domain. Fli-1 and PU.1, related Ets proteins, also bind to the same site. In addition, PU.1 binds to a second site, designated microB, in the Ig heavy-chain enhancer. We demonstrate that the pi binding site is crucial for Ig heavy-chain gene enhancer function. In addition, we show that Erg-3 and Fli.1, but not PU.1, can activate a reporter construct containing a multimer of protein-binding sites, synergistically with helix-loop helix protein E12. We discuss how combinatorial interactions between members of the helix-loop-helix and Ets families may account for the tissue specificity of these proteins. PMID- 8413306 TI - Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between C/EBP and NF-kappa B. AB - The interleukin-8 promoter is transcriptionally activated by interleukin-1, tumor necrosis factor alpha, phorbol myristate acetate, or hepatitis B virus X protein through a sequence located between positions -91 and -71. This region contains an NF-kappa B-like and a C/EBP-like binding site. We show here that several members of the NF-kappa B family, including p65, p50, p52, and c-Rel, can bind to this region, confirming an authentic NF-kappa B binding site in the interleukin-8 promoter. Further, C/EBP binds only weakly to the interleukin-8 promoter site. Electrophoretic mobility shift assays with proteins overexpressed in COS cells and with nuclear extracts from tumor necrosis factor alpha-stimulated HeLa cells demonstrated a strong cooperative binding of C/EBP to its site when NF-kappa B is bound to its adjacent binding site. Transfection studies lead to a model that suggests a highly complex regulation of interleukin-8 gene expression at multiple levels: independent binding of C/EBP and NF-kappa B to their respective sites, cooperative binding of C/EBP and NF-kappa B to DNA, and positive synergistic activation through the C/EBP binding site and inhibition through the NF-kappa B binding site by combinations of C/EBP and NF-kappa B. Thus, the ultimate regulation of interleukin-8 gene expression depends on the ratio of cellular C/EBP and NF-kappa B. PMID- 8413307 TI - Agonist-induced phosphorylation of the luteinizing hormone/chorionic gonadotropin receptor expressed in a stably transfected cell line. AB - Much of the definitive work on G-protein-coupled receptor phosphorylation and its impact on receptor function has been performed with the catecholamine receptors. Evidence for receptor phosphorylation is lacking, however, for G-protein-coupled receptors that bind larger ligands, such as LH/CG. Using immunoprecipitation techniques and a clonal cell line stably transfected with the LH/CG receptor, we show here for the first time that exposure of cells to hCG induces phosphorylation of its cognate receptor. The hCG-induced increase in receptor phosphorylation requires receptor activation because it cannot be elicited with a hCG antagonist and is mediated at least in part by the cAMP second messenger system. This hypothesis is supported by the finding that the hCG-induced receptor phosphorylation is greatly reduced (but not abolished) in a cell line that overexpresses cAMP phosphodiesterase and that receptor phosphorylation can be induced by activation of endogenous cAMP synthesis with prostaglandin E2 or by addition of 8-bromo-cAMP. Last, we show that LH/CG receptor phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. We also examined a potential correlation between LH/CG receptor phosphorylation and uncoupling of the receptor from its effector. Although the phorbol ester-induced phosphorylation of the LH/CG receptor can be correlated with uncoupling, other experiments indicate that hCG-induced uncoupling of the LH/CG receptor can occur under conditions where the cAMP-mediated receptor phosphorylation is greatly reduced (or abolished). PMID- 8413308 TI - Positive regulation of the vitamin D receptor by its cognate ligand in heterologous expression systems. AB - Hormonal vitamin D3 is a major regulator of calcium metabolism and is involved in basic cellular processes, such as those of proliferation and differentiation. These actions are mediated via an intracellular vitamin D3 receptor (VDR), which is a member of the evergrowing steroid hormone receptor superfamily. The interaction between the vitamin D3 ligand and its receptor is thought to be through a classic steroid hormone mechanism. It is notable, however, that 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3] has also been documented as an agent that directly up-regulates endogenous VDR in both intact animals and cultured cells. In this report, we confirm that the levels of recombinantly expressed VDR produced in transiently transfected COS-1 cells also increase several-fold when the cells are treated with 1,25-(OH)2D3. Additionally, we show that a similar pattern is exhibited in a Saccharomyces cerevisiae expression system. This indicates that the mechanism for VDR up-regulation is conserved in both yeast and mammalian cells. Our results show that up-regulation by 1,25-(OH)2D3 is specific to the VDR, and that 1,25-(OH)2D3 does not affect other expressed receptor proteins, such as those for estrogen and progesterone. Finally, we demonstrate that the mechanism of up-regulation apparently occurs at the level of the protein and is most likely due to altered stability of the occupied receptor. Our observations lead us to propose that in addition to the classically viewed role of hormone in receptor activation, 1,25-(OH)2D3 may serve to amplify signal response via homologous up-regulation. PMID- 8413309 TI - Characterization of the mouse FTZ-F1 gene, which encodes a key regulator of steroid hydroxylase gene expression. AB - The cytochrome P450 steroid hydroxylases are coordinately regulated by steroidogenic factor 1 (SF-1), a protein expressed selectively in steroidogenic cells. Based on its expression in steroidogenic tissues and DNA-binding specificity, we isolated a putative SF-1 cDNA from an adrenocortical cDNA library. As evidence that this cDNA encodes SF-1, we now show that it is selectively expressed in steroidogenic cells, that an antiserum against its protein product specifically abolishes the SF-1-related gel-shift complex, and that its coexpression increases promoter activity of the 21-hydroxylase 5' flanking region in transfection experiments. Sequence analyses of the SF-1 cDNA revealed that it is the mouse homolog of fushi tarazu factor I (FTZ-F1), a nuclear receptor that regulates the fushi tarazu homeobox gene in Drosophila. A second FTZ-F1 homolog, embryonal long terminal repeat-binding protein (ELP), was recently isolated from embryonal carcinoma cells. SF-1 and ELP cDNAs are virtually identical for 1017 base pairs, including putative DNA-binding domains, but diverge at their 5'- and 3'-ends. One genomic clone contained both SF-1- and ELP-specific sequences, confirming their origin from a single gene. Characterization of this gene defined shared exons encoding common regions and alternative promoters and 3'-exons leading to differences between the two FTZ-F1 transcripts. We used in situ hybridization with transcript-specific probes to study the ontogeny of SF-1 and ELP expression. ELP transcripts were not detected from embryonic day 8 to adult, consistent with its previous isolation from embryonal carcinoma cells and its postulated role in early embryonic development.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413310 TI - A point mutation in the second zinc finger of the DNA-binding domain of the androgen receptor gene causes complete androgen insensitivity in two siblings with receptor-positive androgen resistance. AB - We have analyzed the nucleotide sequence of complementary and genomic DNAs of the human androgen receptor (AR) gene in two siblings (patients 9006 and 9030) with receptor-positive complete androgen insensitivity (Rec(+)-CAI). Northern analysis indicated that mRNA of the AR was normal in size. However, its expression was relatively reduced in both patients. Consistent with the normal androgen-binding capacity (496 and 552 fmol/mg DNA for patients 9006 and 9030, respectively) but decreased DNA-binding ability (168 fmol/mg DNA) measured in genital skin fibroblasts, no mutation was found in both N-terminal and ligand-binding domains of the AR. However, a single base substitution (G-->A) was found in the second zinc finger of the DNA-binding domain at nucleotide 2372 of the AR cDNA in both cases. This resulted in the replacement of a highly conserved arginine residue (amino acid 614) by a histidine. When the mutated receptor plasmid was cotransfected into PC-3 cells together with the reporter chloramphenicol acetyltransferase gene, chloramphenicol acetyltransferase activity was not induced by 5 alpha-dihydrotestosterone treatment, confirming that the mutation renders the AR nonfunctional and can, therefore, be held responsible for the clinical features in these patients. These results highlight the importance of Arginine-614 in the second zinc finger of the DNA-binding domain of the AR in the protein-DNA interaction. PMID- 8413311 TI - Induction of antiestrogen resistance in human breast cancer cells by random insertional mutagenesis using defective retroviruses: identification of bcar-1, a common integration site. AB - Duration of response to antiestrogen therapy in metastatic breast cancer is limited due to the development of antiestrogen-resistant tumors. The mechanisms involved are not understood but could originate from (epi)genetic alterations within the tumor cells. We have applied in vitro random insertional mutagenesis with replication defective retroviruses to identify those genes playing a key role in development of antiestrogen resistance in human breast cancer cells. Eighty antiestrogen-resistant cell clones were isolated from 7 x 10(8) estrogen dependent ZR-75-1 cells, mass-infected with defective retroviruses and subjected to 4-OH-tamoxifen selection. Integration site-specific DNA probes were made by inverse polymerase chain reaction techniques and used to search for common integration sites. Six cell clones were identified with retroviral genome integrations in the same orientation in a single locus, designated breast cancer antiestrogen resistance locus-1 (bcar-1). These bcar-1 cell clones had lost estrogen receptor expression and had become estrogen independent. Our results strongly suggest that alteration of the bcar-1 locus is responsible for development of antiestrogen resistance in human breast cancer cells in vitro. In addition, we have shown that in vitro insertional mutagenesis using defective retroviruses can be applied for gene tagging in human cells. PMID- 8413312 TI - Involvement of the coding sequence for the estrogen receptor gene in autologous ligand-dependent down-regulation. AB - The estrogen receptor (ER) appears to be down-regulated by its own ligand in some estrogen (E2)-responsive tissues as well as in cell lines such as MCF-7 and GH3. Surprisingly, we observed ER down-regulation in a newly constructed E2-responsive cell line (Rat1 + ER), in which expression of the coding region of the ER cDNA was driven by the Moloney murine leukemia virus long terminal repeat. We present evidence that the coding region of the ER cDNA, but not the Moloney murine leukemia virus long terminal repeat, possesses a sequence(s) necessary for ER down-regulation. The observed down-regulation occurs at ligand-binding and protein, as well as mRNA, levels. Marked decreases in both protein and mRNA levels were observed as early as 3 h after E2 treatment. Furthermore, maximal down-regulation occurred by 18-24 h with ligand-binding, and mRNA levels reached approximately 20% that of controls. ER down-regulation in Rat1 + ER cells is only partially inhibited by the presence of cycloheximide and therefore suggests a direct participation of the ER in this process. E2 does not appear to influence the stability of the ER transcript, which implies that negative regulation is occurring at the transcriptional level. Finally, since we can demonstrate ER binding to a portion of the cDNA sequence, we propose a mechanism whereby ER binding to its putative negative element leads to transcriptional repression of the upstream promoter. PMID- 8413313 TI - Basic fibroblast growth factor is a testicular germ cell product which may regulate Sertoli cell function. AB - Previously, a proteinacious factor secreted by a mixture of rat testicular spermatocytes and round spermatids was shown to stimulate transferrin mRNA and protein levels in Sertoli cells. To identify the germ cell-secreted proteins which affect Sertoli cell functions, concentrated germ cell-conditioned medium was fractionated by reverse-phase HPLC. The fraction which eluted at 35% acetonitrile increased transferrin secretion in Sertoli cell cultures 2.4-fold above the basal level. Both the active fraction and a protein extract from cultured germ cells were positive for basic fibroblast growth factor (bFGF) as determined by Western blot analysis and immunoprecipitation. The apparent molecular sizes of the immunoreactive proteins were 30, 27, and 24 kilodaltons (kDa). By immunohistochemistry, bFGF was shown to be present in pachytene spermatocytes and Leydig cells. The bFGF receptor was also examined by immunohistochemistry and found to be present in Leydig cells, round and elongated spermatids, and Sertoli cells. The presence of receptors was more pronounced in stages I-VIII. Western blot analysis confirmed that the receptors were expressed in isolated round spermatids, elongated spermatids, and Sertoli cells. Two major receptor species with apparent molecular sizes of 120 and 145 kDa were detected in the rat testis. Germ cells contained both of these receptors, but Sertoli cells possessed only the 120-kDa receptor. From these experiments, it is evident that bFGF is a germ cell product which may regulate Sertoli cell function. The expression of bFGF and its receptor may be an important component of germ cell Sertoli cell and/or germ cell-germ cell communication during spermatogenesis. PMID- 8413314 TI - Gonadal steroid hormone regulation of human and mouse follicle stimulating hormone beta-subunit gene expression in vivo. AB - Transgenic mice that express a 10-kilobase human FSH beta (hFSH beta) gene exclusively in pituitary gonadotropes were used to study the regulation of hFSH beta gene expression by gonadal steroids. For comparison, the mouse FSH beta (mFSH beta) gene was studied in parallel in nontransgenic sibling (normal) mice. The hFSH beta gene showed a sexually dimorphic expression pattern, identical to mFSH beta, in the mouse environment. Intact normal and transgenic male mice had elevated (P < 0.05) levels of serum [16 +/- 2 ng/ml (normal); 38 +/- 6 (transgenic)] and pituitary FSH content [2 +/- 0.3 micrograms/mg protein (normal); 36 +/- 6 (transgenic)] and FSH beta mRNA [1.47 +/- 0.10 arbitrary density units (normal); 1.00 +/- 0.23 (transgenic)] compared to the corresponding female mice ([< 2.0 ng/ml (normal and transgenic)] [0.1 +/- 0.01 microgram/mg protein (normal); 0.2 +/- 0.03 (transgenic)] [< 0.03 arbitrary density units (normal and transgenic)]). Serum FSH levels were increased (P < 0.05) 2 weeks after castration of normal (22 +/- 2 ng/ml) and transgenic males (135 +/- 19 ng/ml) and were suppressed (P < 0.05) by testosterone [7 +/- 0.8 ng/ml (normal); 12 +/- 2 (transgenic)] or estradiol [14 +/- 1 ng/ml (normal); 16 +/- 1 (transgenic)] replacement. The increased serum FSH levels were associated with an inverse drop (P < 0.05) in pituitary FSH content to 1 +/- 0.1 microgram/mg protein in normal and 16 +/- 2 in transgenic males. Testosterone replacement further suppressed (P < 0.05) pituitary FSH content in transgenic (3 +/- 0.5 micrograms/mg protein) but not normal (1 +/- 0.1) males.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413315 TI - Glucocorticoid receptor binding site in the mouse alpha-amylase 2 gene mediates response to the hormone. AB - Amylase gene expression has been shown to be positively regulated by glucocorticoids. Previous reports have suggested that this effect is indirect. We have addressed this question in a mouse exocrine pancreas cell line, 266-6, in which basal level of expression of amylase mRNA is low but inducible by glucocorticoids. In these cells the effect of glucocorticoids is not inhibited by cycloheximide at early time points. Reporter plasmids containing 224 base pairs of mouse amylase 5'-flanking DNA are positively regulated by glucocorticoids in gene transfer experiments. Glucocorticoid receptor purified from rat liver binds to the amylase promoter from position -56 to -33 and at the start of transcription. Site-directed mutation at the upstream position (-47 to -42) eliminates response to glucocorticoids in transient gene transfer experiments. Thus, glucocorticoid regulation of the mouse amylase gene is a direct effect and is mediated via a receptor binding site in the promoter region of the gene. Inhibition of the hormone response by cycloheximide at later time points after induction suggests the additional requirement for a short-lived factor. The DNA binding domain of the glucocorticoid receptor binds to a single site in the amylase promoter as a monomer, suggesting that both receptor binding sites as well as an additional short-lived factor are required to obtain induction. PMID- 8413316 TI - The Ras and protein kinase C signaling pathways are functionally antagonistic in GH4 neuroendocrine cells. AB - Oncogenic Ras appears to act via protein kinase C (PKC)-dependent and PKC independent pathways. In several systems, oncogenic Ras cooperates with c-Jun to activate gene transcription from promoters containing an AP-1 site by augmenting phosphorylation of the transcriptional activation domain of c-Jun. We have previously shown that oncogenic valine 12 Ras and PKA each separately activate the rat PRL (rPRL) promoter but together are mutually antagonistic. The goal of this study was to determine whether oncogenic Ras acts through PKC and c-Jun to activate transcription of an rPRL-luciferase reporter construct transiently transfected into GH4 rat pituitary cells. Our results show that phorbol 12 myristate 13-acetate (TPA) activates rPRL promoter activity through PKC, and that TPA activation of PKC diminishes the Ras response in a dose-dependent manner. Additionally, inhibition of PKC with staurosporine does not block the oncogenic Ras effect. Similarly, rPRL promoter activity in GH4 cells expressing oncogenic Ras fails to respond to TPA activation of PKC. Finally, cotransfection of a c-Jun expression vector results in inhibition of basal, TPA, and oncogenic Ras stimulated activity of the rPRL promoter. Thus, we show that the mechanism of Ras signaling does not involve PKC, and that PKC does not signal via Ras. Taken together, these results verify that the Ras and PKC signaling pathways are separate and mutually antagonistic, and that c-Jun is not the nuclear mediator of either the Ras or PKC signal. These findings emphasize the possibility that the roles and/or functions of specific components in signaling pathways may be different in distinct cell types. PMID- 8413317 TI - Transcriptional and posttranscriptional regulation of human androgen receptor expression by androgen. AB - Autoregulation is a control mechanism common to several proteins of the steroid/thyroid hormone receptor superfamily. In this work, the effect of androgens and antiandrogens on the expression of the human androgen receptor (hAR) in prostate and breast cancer cell lines was studied. Northern blot analysis revealed a decrease in hAR steady state RNA levels in LNCaP cells by 3.3 nM of the synthetic androgen mibolerone. Maximal down-regulation of hAR RNA to 30% of control levels occurred 48 h after hormone addition. T47D breast cancer cells showed a similar effect with mibolerone, while hAR expression in normal skin fibroblasts did not respond to androgen treatment. As shown by nuclease S1 analysis, hAR transcripts initiate at three principal start sites, all of which are equally sensitive to androgen. Steroidal as well as nonsteroidal antiandrogens were capable of partially antagonizing androgen-mediated hAR RNA down-regulation in LNCaP and T47D cells, while not exerting a significant effect when administered alone. While hAR RNA stability was increased by hormone, nuclear run-on analysis revealed a 4-fold reduction of hAR gene transcription 96 h after androgen treatment. Although decreased hAR RNA levels did not coincide with a parallel decrease in AR protein levels, analysis of androgen-inducible reporter constructs demonstrated that prolonged androgen administration to cells results in a progressively impaired sensitivity of the intracellular androgen response mechanism. These results show that prolonged androgen exposure leads, besides its effect on hAR RNA levels, to functional inactivation of the AR. Thus, in vivo, posttranslational control of AR activity appears to be a novel mechanism of negative autoregulation of androgen effects on gene expression. PMID- 8413318 TI - The use of anchored polymerase chain reaction for the study of large numbers of human T-cell receptor transcripts. AB - Anchored-PCR (A-PCR) is an approach designed to amplify and clone sequences with unknown 5' or 3' extremities. A-PCR is therefore appropriate for studying variable region of T-cell receptors (TCRs) expressed in polyclonal T-cell populations since it does not prejudge which variable gene segments are actually being used. We report here some critical modifications in the initial procedure to make it easy to clone and sequence large series of TCR transcripts. They have been introduced to improve both the yield and specificity of TCR amplified products and include re-amplification, size selections of the material combined with the successive use of nested TCR constant region specific primers. This procedure has been successfully applied to the study of the repertoire of both TCR alpha/beta+ and gamma/delta+ human T-cells. The efficiency of the present A PCR protocol will help to precisely analyze TCR usage in normal and pathological situations. PMID- 8413319 TI - The mouse mammary tumor virus long terminal repeat encodes a 47 kDa glycoprotein with a short half-life in mammalian cells. AB - The mouse mammary tumor virus proviral DNA contains an open reading frame in the 3' long terminal repeat which can code for a 36 kDa polypeptide with a putative transmembrane sequence and five N-linked glycosylation sites. This gene is known to code for a superantigen which deletes a specific subset of CD4+ T lymphocytes in vivo. The superantigen encoded by the exogenous mouse mammary tumor virus of the GR strain acts specifically on V beta 14 bearing T cells. We produced recombinant vaccinia viruses to express either the complete or a truncated ORF protein after infection of primate cells in culture. The complete ORF gene in mammalian cells leads to the production of a 47 kDa protein which is specifically detected with an anti-ORF-peptide antiserum. The 47 kDa protein can be labeled with D-[2-3H]mannose and its synthesis is inhibited by tunicamycin, an N-linked glycosylation inhibitor, indicating that it is a glycoprotein. The truncated ORF protein beginning at the second ATG of the open reading frame is also modified, but the C-terminal half of ORF, starting at the fifth ATG, has the expected size of the non modified polypeptide. Pulse-chase experiments indicate that the ORF protein has a short half-life of about 1.5-2 hr. PMID- 8413320 TI - HIV-1 gp41 binds to several proteins on the human B-cell line, Raji. AB - Based on our findings that HIV-1 gp41 independently of CD4, can bind to the helper T-cell line H9, we characterized putative binding of HIV-1 gp41 to B-cell lines, Raji, Bjab and Ramos. Using fluorescence-activated cell sorter (FACS) we examined the binding of soluble gp41 (sgp41; Env amino acid 539-684) to these B cell lines. Using sgp41 attached to sepharose beads Raji cell lysates were absorbed. The sgp41-eluate of Raji cell lysates could inhibit the sgp41-binding to Raji cells. By SDS-PAGE of sgp41-eluate of Raji cell lysates four strong protein band, 37, 45, 49 and 62 kD, and a weak band of 92 kD were stained with Coomassie blue. By Western blot (ligand blot) analysis using sgp41 four protein bands, 37, 45, 49 and 62 kD, were observed in sgp41-eluate of Raji cell lysates. To test the individual proteins the five proteins were isolated from the sgp41 eluate of Raji cell lysates. Three proteins, 45, 49 and 62 kD, each could partially inhibit the sgp41-binding to Raji cells. The results suggest that these three proteins in Raji cell lysates are possible candidates for the putative gp41 receptor(s). PMID- 8413321 TI - The immunochemistry of sandwich ELISAs--VI. Greater than 90% of monoclonal and 75% of polyclonal anti-fluorescyl capture antibodies (CAbs) are denatured by passive adsorption. AB - Quantitative data are presented showing that the method most commonly used to immobilize antibodies in microtiter immunoassays functionally inactivates most of the antibodies. These results were collected using five affinity purified polyclonal antibodies (pAbs) and six monoclonal antibodies (mAbs) specific for fluorescein (FLU) as capture antibodies (CAbs). These CAbs were tested for their ability to capture FLU4.2-BSA after immobilization by passive adsorption, the Protein-Avidin-Biotin-Capture (PABC) system or using previously adsorbed anti globulins. Results indicate that under optimal conditions, < 10% of monoclonal capture antibody equivalents (CAbeqv) and congruent to 22% of polyclonal CAbeqv remain functional after passive adsorption. Immobilization via the PABC system improved the performance of mAbs by more than five-fold but had less than a two fold effect on pAbs. Many CAbs immobilized using an anti-globulin retained full activity including the ability to bind two molecules of FLU4.2-BSA/molecule of CAb. The latter result is not necessarily a recommendation for the use of anti globulin immobilization, since the number of functional CAbeqv per well is not significantly greater than that which can be achieved using passive adsorption. PMID- 8413322 TI - Post-translational processing of a major histocompatibility complex-encoded proteasome subunit, LMP-2. AB - Proteasomes are abundant, multisubunit protein complexes found in the cytoplasm and nucleus of eukaryotic cells that catalyze both ubiquitin-dependent and ubiquitin-independent protein degradation. In addition to their role in normal protein turnover, proteasomes are believed to be involved in the production of most antigenic peptides presented to T cells by major histocompatibility complex (MHC) class I molecules. A distinct subset of mouse proteasomes contain a subunit called LMP-2, which is encoded within the MHC. Here we demonstrate that a previously isolated proteasome cDNA clone encodes the LMP-2 subunit, and that two distinct forms of this subunit may be found in the proteasome complex. One form probably corresponds to the primary translation product, whereas the second form appears to be post-translationally processed by removal of the amino-terminal 20 amino acids. Determination of the location of intron/exon boundaries in the Lmp-2 gene indicated that these residues correspond precisely to the first exon of the gene. PMID- 8413323 TI - The murine NK2.1 antigen: a 130 kD glycoprotein dimer expressed by a natural killer cell subset of the spleen, thymus and lymph nodes. AB - Murine natural killer (NK) cells express a few antigens not found on other leukocyte subsets. The NK1.1 antigen, that is present in only a few mouse strains, has been extensively characterized whereas our knowledge of the NK2.1 antigen, which is more commonly expressed, remains, as yet, limited. Our laboratory has previously reported the production of a mAb (4LO3311) recognizing a murine NK cell-specific molecule with a similar strain distribution as the NK2.1 antigen formerly defined with an NZB anti-BALB/c antiserum. In this study, we demonstrate by sequential immunoprecipitation that 4LO3311 represents the first NK2.1 antigen-specific mAb. This reagent was used to immunoprecipitate the NK2.1 antigen from 125I-labeled lysates of fresh NK-enriched spleen cells. SDS PAGE analyses revealed that the NK2.1 antigen is expressed at the cell surface as a N-glycosylated disulfide-linked protein dimer with approximately 65 kD subunits. The NK2.1 antigen is likely to be anchored in the plasma membrane by a peptide moiety since its expression on NK cells was not affected by treatment with phosphatidylinositol-specific phospholipase C. In addition to be present on a splenic NK cell subset, the NK2.1 antigen is shown to be expressed by a small number of CD4-CD8-thymocytes and by a subset of CD4-CD8-IgG- lymph node cells. Finally, it is shown here that unlike the NKR-P1, the rat homologue of the murine NK1.1 antigen, neither the NK2.1 nor the NK1.1 antigen is expressed by polymorphonuclear leukocytes. PMID- 8413324 TI - Sequence of ovine Ig gamma 2 constant region heavy chain cDNA and molecular modelling of ruminant IgG isotypes. AB - Ovine mesenteric lymph node mRNA was used for PCR amplification of DNA coding for immunoglobulin gamma 1 and gamma 2 heavy chain constant regions. Primers complementary to regions of CH1 conserved between ruminants were used for upstream priming, with downstream priming on the poly-A segment. PCR products of the appropriate length were cloned and gamma positive clones selected with a CH1 conserved-region probe. Of these, gamma 1 clones were positively selected and gamma 2 clones negatively selected with a gamma 1 hinge-specific probe. Ovine gamma 2 cDNA has 93% identity of nucleotides with ovine gamma 1. Both ovine gamma 1 and gamma 2 CH1 domains encoded two consecutive cysteine residues (Cys-127, 128, Kabat numbering), an arrangement which is deduced to form a pair of disulphide bridges, one to the L chain and one as an intra-chain bridge to the uppermost Cys of the hinge, as in rabbit and goat IgG. The majority of the differences between the isotypes occur in the hinge region and an evolutionary pattern for ruminant IgG hinges can now be identified. IgG1 isotypes are typical, with hinges containing the C-terminal Cys-Pro motif, but deletion and replacement of nucleotides (in the ancestral gene) of ruminant gamma 2 has shortened the IgG2 hinge, removing the Cys-Pro motif and the consensus high affinity Fc gamma RI receptor motif at the start of CH2. An N-terminal glycosylation site and the peptide motif for complement C1q binding are present in CH2 of both isotypes. The hinge regions of gamma 1 and gamma 2 and predicted structures for ovine IgG1 and IgG2 have been modelled. Close apposition of Fab and Fc in IgG2 produces steric hindrance at the normally accessible Fab/hinge/Fc interface; the structural differences between the ruminant isotypes form a basis for understanding some of the differences in their effector properties. PMID- 8413325 TI - Post-infectious human serum antibodies inhibit IgA1 proteinases by interaction with the cleavage site specificity determinant. AB - Bacterial pathogens of the genera Neisseria and Haemophilus secrete IgA1 proteinases which cleave human IgA1 in the heavy chain hinge region. The exact peptide bond cleaved is strain-dependent, but remains invariant despite repeated subculture. Haemophilus influenzae and Neisseria meningitidis produce proteinases of two cleavage site specificities (type 1 and type 2). We examined serial acute and convalescent sera from patients recovering from meningitis due to N. meningitidis or H. influenzae, and found a significant rise in serum titer of inhibitory antibodies against these enzymes. In each case the proteinase from the infecting organism was more susceptible to inhibition than were proteinases from that genus that had different cleavage specificity. Inhibition of sixteen type 1 type 2 hybrid H. influenzae IgA1 proteinases revealed complete concordance between inhibitory titer and cleavage site specificity. Inhibition of hybrid proteinases differing in a 123 amino acid segment known to determine cleavage site specificity (termed the CSD) further localized the site of antibody action to this site. These results from a limited number of patients with natural infections suggest that inhibiting antibody recognizes epitopes within the CSD. Alternatively, antibody may bind to epitopes outside the CSD and inhibit via steric hindrance. PMID- 8413326 TI - Evidence for antigen driven selection in two monoclonal auto-antibodies derived from nonobese diabetic mice. AB - The nonobese diabetic (NOD) mouse is a model of human type I diabetes. This diabetes is due to massive infiltration of the pancreatic beta cell of islets by autoreactive T cells (insulitis) followed by the destruction of insulin-producing cells. Circulating autoantibodies are also detected, notably against glutamic acid decarboxylase, peripherin and insulin. Two monoclonal autoantibodies directed against insulin and peripherin were obtained by fusing NOD spleen and myeloma cells. We report here the nucleotide sequence of the genes encoding for the V regions of these two antibodies. Somatic mutations were identified by comparing the light chain nucleotide sequence of one of these autoantibodies with its germline counterpart precursor established from NOD mice after PCR gene amplification. The other one displays N additions on both sides of the D region. These results strongly suggest that both autoantibodies have undergone diversification, either N additions or somatic mutations, and therefore present structural features of antibodies derived from animals immunized against exogenous antigens. PMID- 8413327 TI - Rat IgG subclasses mediating binding and phagocytosis of target cells by homologous macrophages. AB - Attachment and ingestion of 51Cr-labelled TNP-SRBC sensitized by rat IgG1, IgG2a or IgG2b-type antibodies by homologous, elicited peritoneal macrophages were studied. IgG1 was found to be the most efficient isotype in mediating these functions. The antibody doses required for a significant attachment were found to differ with the isotype of Ab, while doses needed for a significant phagocytosis and antibody-dependent cellular cytotoxicity (ADCC) varied between 400-700 Ab/SRBC with all the isotypes studied. Both binding and phagocytosis were also influenced by the degree of hapten conjugation when target cells were sensitized by IgG1. Inhibition of these functions by soluble immune complexes and monomeric immunoglobulins suggests the involvement of two Fc gamma R in binding of the three isotypes. Based on the present work and on previous results we conclude that IgG2a interacts with a receptor binding complexed IgG only (Fc gamma RII), IgG2b binds to a different receptor which appears to bind monomeric ligand as well (Fc gamma RI), while IgG1 seems to interact with both types of receptor. We propose that phagocytosis can be mediated by both Fc gamma RI and Fc gamma RII. PMID- 8413328 TI - The interaction between different domains of staphylococcal protein A and human polyclonal IgG, IgA, IgM and F(ab')2: separation of affinity from specificity. AB - Binding properties of staphylococcal protein A (SpA) to different human immunoglobulins have been investigated. In this analysis, intact SpA as well as SpA-derived fragments containing one to five IgG-binding domains of different compositions, were used. The affinity binding constants of the different proteins to human polyclonal IgG, IgA, IgM and F(ab')2-fragments as well as their binding capacity to the immunoglobulin molecules were determined. The results show that although all the proteins bound to IgG, regardless of size or composition, the binding strength differed significantly. Proteins containing five domains have a stronger affinity for IgG than those containing one or two. There were no marked differences in binding strength between different domains. However, the binding ability to IgA and IgM showed a marked difference between the various SpA-derived proteins of different compositions. This discrepancy was correlated to differences in their relative binding properties to isolated F(ab')2-fragments of IgG. Hence, we conclude that the binding affinity is mainly affected by the number of domains, whereas the binding specificity is to a large extent determined by which domains are selected. PMID- 8413329 TI - Tumor necrosis factor alpha mediates a T cell receptor-independent induction of the gene regulatory factor NF-kappa B in T lymphocytes. AB - We investigated the molecular basis of the ability of DCEK experimental antigen presenting cells (APCs) to induce the nuclear form of the transcription factor NF kappa B in T lymphocytes without engagement of the T cell receptor. We found that NF-kappa B induction did not require contact between the APCs and T lymphocytes and could be achieved by medium conditioned by the APCs. The APCs were found to express low levels of mRNA for TNF alpha. The addition of antibody against TNF alpha blocked the ability of APCs to induce NF-kappa B. These observations were extended by the finding that NF-kappa B was also induced in T lymphocytes separated by a membrane from a mixture of T lymphocytes, splenic APCs and antigen by a TNF alpha-dependent mechanism. Together, these findings suggest that induction of NF-kappa B in antigenically stimulated or 'bystander' T cells may take place through stimulation by TNF alpha as well as in response to T cell receptor occupancy. PMID- 8413330 TI - IL-2 regulates the expression of the NK-TR gene via an alternate RNA splicing mechanism. AB - We have recently isolated and characterized human and mouse genes of a putative natural killer (NK) cell tumour-recognition protein (NK-TR) that is specifically expressed in NK cells. This gene codes for a 150 kD protein with a cyclophilin related amino terminus followed by several positively charged domains. We report here the discovery of two sites of alternate splicing in the 5' region of the NK TR mRNA. One of these events caused a frameshift in the open reading frame by splicing in a 28 bp exon within the cyclophilin coding region, resulting in the premature termination of the NK-TR protein. The second alternate splice stemmed from the use of an internal splice acceptor within an exon, producing a deletion of 25 amino acids in the NK-TR protein. The activation of NK cells by IL-2 produced a change in the splicing pattern that resulted in increased production of mRNAs capable of producing the complete NK-TR protein. PMID- 8413331 TI - Molecular cloning and characterization of a novel neutrophil chemotactic factor from a filarial parasite. AB - To compare the molecular structure of a parasite-derived neutrophil chemotactic factor (NCF) with host-derived NCFs or other NCFs, molecular cloning of cDNA encoding NCF derived from Dirofilaria immitis adult worm (DiNCF) was performed. A D. immitis cDNA library was screened with an antibody to DiNCF, and one DiNCF cDNA clone (pD-4) was isolated. A fusion protein of pD-4 and gene 10 protein showed significant neutrophil chemotactic activity whereas gene 10 protein itself showed marginal neutrophil chemotactic activity. The total nucleotide sequence analysis revealed that pD-4 was 994 bp long with a 432 bp open leading frame encoding a 143 residue protein. The NH2-terminal amino acid sequence of the natural DiNCF and the deduced amino acid sequence from the cDNA showed that the mature functional protein was comprised of 112 amino acids. Although the deduced amino acid sequence of this protein did not show overall homology to host-derived NCFs or other known proteins, it contained a similar sequence (Met-Phe-Lys) to the known chemotactic peptides. The possibility of the functional epitope of DiNCF is discussed. PMID- 8413332 TI - [Apnea in premature infants. Epidemiology, pathophysiology and possibilities for prevention]. AB - The pathogenesis of apnoea in preterm infants is poorly understood. Thus, it is yet unknown (1) whether the descriptive distinction between "central", "obstructive" and "mixed" apnoeas is also reflected by differences in their respective pathogenesis, (2) how and where the airway closure occurs during the so-called "obstructive" apnoeas, (3) whether "central" apnoeas do indeed result from a disturbance in the central nervous control of breathing, as implied by their name, or whether they constitute a reflex response to a disturbance in the periphery of the lung, and (4) whether non-apnoeic mechanisms contribute to the hypoxaemia so frequently observed during these episodes. This paper summarises the present knowledge regarding the above issues. It also reviews the current definitions for apnoea, bradycardia and hypoxaemia in preterm infants, describes indications for intervention, and discusses some strategies aimed at preventing these episodes. PMID- 8413333 TI - [Periventricular leukomalacia]. AB - The present knowledge on periventricular leucomalacia is reviewed. Its incidence is about 10% in preterms infants < 32 gestational weeks. By careful ultrasonographic scanning four stages can be distinguished: Prolonged flares, resolution, cystic leucomalacia, ventriculomegaly. Periventricular leucomalacia (PVL) is much more correlated with later neurologic impairment than intraventricular haemorrhage (apart from grade IV). PVL is caused by ischemia of the periventricular white matter mainly due to arterial hypotension and disturbed regulation of cerebral blood flow. The interactions between blood pressure, pCO2, pO2 and cerebral blood flow are complex and poorly understood. Regarding the clinical management it seems important to avoid hypotension, fluctuating pCO2 and hypoxemia/anemia in order to prevent white matter ischemia. Research should be focussed on the development of monitoring methods (doppler sonography, near infrared spectroscopy, time compressed electroencephalography), which allow to detect ischemic events to prevent neurologic impairment in about 1000 preterm infants per year in Germany. PMID- 8413334 TI - [Fatal course of a sequestration crisis in hemoglobin SC disease]. AB - A 14 year-old patient developed severe anemia and splenomegaly 2 days after the onset of a febrile upper airway infection. The hemoglobin concentration had dropped to 1.1 g/dl. Death occurred as consequence of the acute anemia and peripheral circulatory failure. The crisis was caused by an acute splenic sequestration. Hemoglobin SC disease could be identified as the underlying disorder. Hemoglobin SC disease usually has a milder course than sickle cell disease. However the patients may develop the same crisis-like symptoms. Splenic sequestration is caused by the occlusion of the splenic sinuses due to sickled and aggregated erythrocytes with subsequent trapping of large blood volumes and circulatory failure. Regular transfusions and/or splenectomy are recommended to prevent splenic sequestration crisis. PMID- 8413335 TI - [Williams-Beuren syndrome in combination with celiac disease]. AB - At age 5 years coeliac disease was diagnosed in a boy with Williams-Beuren syndrome. Gastrointestinal symptoms may be similar in both disorders. Short stature, not only in comparison to the normal population but also in comparison to syndrome-specific growth standards, and failure to thrive led to the diagnosis coeliac disease. During the first year of dietetic therapy the boy showed marked weight gain, catch-up growth and a normalisation of red blood count. Gliadin-IgG antibodies and endomysium-IgA antibodies who were positive before treatment were no longer detectable. Our case demonstrates that syndrome-specific growth standards are helpful even though the combination of Williams-Beuren syndrome with a second disorder is very unlikely (Williams-Beuren syndrome and coeliac disease are to be expected in 1:13.7-1:25 millions). PMID- 8413336 TI - [Ultrasound diagnosis of birth trauma lesion of the cervical spine]. AB - Ultrasonographic imaging in the newborn via a posterior approach allows the visualization of the craniocervical junction. We describe a case presenting with clinical signs of a spinal cord injury after delivery by forceps extraction. The sonographic examination demonstrated an increased cervical cord echogenicity during the first days of life, and a decrease of cord size at the craniocervical junction after 3 weeks. MRI confirmed these findings. Sonography is useful in showing severe spinal cord injury with minimal handling of these severely traumatized neonates. PMID- 8413337 TI - [Cardiac symptoms in 2 patients with Seckel syndrome]. AB - Seckel's Syndrome is a rare (< 1:10,000) malformation, presumably inherited as an autosomal-recessive trait. It is characterized by failure to thrive, mental retardation, bird-like malformation of the face and further abnormalities. Cardiac anomalies have not been described so far. We report two patients with cardiac malformations, a complex defect and a patient ductus arteriosus. PMID- 8413338 TI - [Hemorrhagic shock and encephalopathy syndrome]. AB - BACKGROUND: There are only a few case reports from Germany of hemorrhagic shock and encephalopathy syndrome (HSES), whereas the incidence of this entity is probably much higher than reported. Thus, it seems necessary to describe further observations of patients with HSES. METHODS: Seven patients (3 boys, 4 girls, aged 8 days to 9.5 months) were observed between 1987 and 1992, who fulfilled the diagnostic criteria of HSES. Their clinical data and laboratory findings were evaluated. RESULTS: Diarrhoea preceded onset of HSES in 4 patients, 3 patients had loose stools immediately after admission. Disturbed consciousness, severe shock, metabolic acidosis, fever and disturbed coagulation parameters were present in all of them. In 2 patients low concentrations of alpha-1-antitrypsin were found during the acute phase of HSES. Edema of the brain was diagnosed in 4 patients, additional cerebral infarction in 2 patients and cerebral hemorrhage in 1 patient. Three patients died, 3 of 4 survivors had severe brain damage. CONCLUSION: HSES is a clinical entity with unknown etiology, acute onset, fulminating course and poor outcome. Prognosis may be improved by thorough treatment of brain edema, shock and disseminated intravascular coagulation. PMID- 8413339 TI - [Ifosfamide-induced nephrotoxicity]. AB - BACKGROUND: A number of acquired De-Toni-Debre-Fanconi-Syndromes have been reported after cytostatic treatment with Ifosfamide. For the majority of these patients prognosis of renal function was poor. In our study we evaluated the frequency of subclinical tubulopathies, the influence of the cumulative Ifosfamide dose and other risk factors and the prognosis of once established tubular dysfunction. METHODS: 79 patients after polychemotherapy regimens employing Ifosfamide (n = 39), Ifosfamide plus Cisplatinum (n = 35) or Cisplatinum (n = 5) were examined at least 3 months after completion of therapy. Beside the creatinine-clearance we evaluated glomerular and tubular function by measurement of transferrin, immunoglobuline G, Alpha-1-microglobuline and N Acetyl-beta-D-Glucosamidaseexcretion and by tubular reabsorption of phosphate and aminoacids. RESULTS: Renal hyperaminoaciduria was found most often, and there was no linear correlation between the cumulative Ifosfamide-dose and phosphate reabsorption. A reduced glomerular filtration rate and glomerular proteinuria were found in about 10.5% of patients. Approximately half of the patients had tubular dysfunction. Impairment of phosphate reabsorption once established did not normalize in the majority of patients. Cisplatinum was found to worsen the Ifosfamide-induced tubulopathy. CONCLUSION: Our results indicate subclinical tubulopathy after Ifosfamide in a high proportion of patients. This Ifosfamide induced nephrotoxicity is worsened by Cisplatinum and seems to be irreversible for the majority of patients. To describe and to prevent Ifosfamide-induced nephrotoxicity, further studies should focus on: 1. the identification of the risk-groups/patients and additional risk factors, 2. the estimation of the prognosis of once established renal damage, 3. the clarification of the pathomechanism. PMID- 8413340 TI - [Necrotizing enterocolitis in the newborn infant. Pathogenetic risk factors in a 3 year analysis]. AB - In a clinical study possible risk factors causing necrotizing enterocolitis (NEC) were studied in 17 preterm infants with NEC and compared with a control group of 17 healthy preterm infants. We found a more frequent rate of blood exchange transfusions and a lower rate of breast milk feeding in patients who later developed NEC than in the control group. But there were no differences between both groups concerning factors promoting hypoxia of the gut, the number of infections, antibiotic treatment, or hyperosmolar enteral feeding. The preterm infants with NEC did not show any characteristics in history, physical examination and clinical course. These children could not be distinguished as a risk group for this illness from the newborn infant control group. Therefore, with the exception of breast milk feeding, no special prevention of the necrotizing enterocolitis can be recommended. PMID- 8413341 TI - [New conservative therapy of phimosis]. AB - A new conservative method for the treatment of phimosis is presented: similar to the labio-synechia of girls a local treatment with an oestrogen containing cream is performed successfully. After approximately three weeks about 90% of the boys could easily retract their prepuce. We hope that in future an operation will be less often necessary because of this successful conservative method. PMID- 8413342 TI - [Adrenogenital syndrome with 21-hydroxylase deficiency]. PMID- 8413343 TI - [Comments on Immune stimulation/-modulation with preparations consisting of extracts of Sun-flower or bacterial lysate]. PMID- 8413344 TI - [Halogen-induced panniculitis caused by potassium bromide]. AB - Necrotizing panniculitis due to potassium-bromide is a drug induced allergic reaction following stimulation of lymphocytes as demonstrated for the first time by a lymphocyte transformation test (LTT). We named the disease "halogen panniculitis" because of similar generally known reactions to iodides and describe the typical symptoms in three own cases. Bromoderma tuberosum tends to be a similar kind of allergy. For the first time pancreatitis to potassium bromide in men has been observed, which has already been described in epileptic dogs treated with potassium bromide. PMID- 8413345 TI - [Hemoglobin SD disease]. AB - A 7 year old Kurdish girl presented with a cerebral infarction and a resulting hemiparesis following a blood transfusion. Examination of the blood film suggested sickle cell anaemia. However a simultaneously carried out haemoglobin electrophoresis showed haemoglobin S and haemoglobin D. The diagnosis of haemoglobin SD disease was thereby established. This kind of haemoglobin disorder usually shows a mild clinical manifestation. According to our knowledge such serious cases have not been published before. With this disease the main emphasis is on the prevention of recurrent cerebral infarctions bay a long-term transfusion programme whereas the effects of the cerebral infarction are treated in the usual way. PMID- 8413346 TI - [Legionellosis in a newborn infant]. AB - We describe a case of successful therapy of a neonatal Legionellosis with Erythromycin. On his 6th day of life a full term newborn with normal body weight was affected by a severe pneumonia. This was at first resistant to therapy and required mechanical ventilation. Diagnosis of Legionella pneumophila serotype 1 was made by culture from bronchial lavage. Only few cases of neonatal Legionellosis have been reported until now. In three cases diagnosis was made post mortem. PMID- 8413347 TI - [Percentile curves of functional residual capacity of newborn infants]. AB - BACKGROUND: Helium dilution or nitrogen washout techniques are commonly used to measure the functional residual capacity (FRC) in newborn babies. For both techniques equipment is available. Its clinical application, however, was difficult since there were no FRC reference ranges for the different weight groups of newborn infants; reference studies are very time consuming and cause several technical and ethical problems. METHODS: Statistical methods were used to evaluate several results from the literature and to exclude significantly differing data of some authors. Measured values of FRC were related to body weight, and the percentiles of FRC/kg were estimated by the smoothed cumulative distribution function of the pooled data. RESULTS: No significant differences were found between the results of helium dilution and nitrogen washout techniques. The distribution of 178 results showed a significant skewedness (x5% = 18.3 ml/kg; x50% = 27.5 ml/kg x95% = 42.8 ml/kg), which could also be seen in the percentile curves calculated approximately, assuming a linear development of functional residual capacity with body weight. CONCLUSION: Percentile curves describe best the development and interindividual variability of FRC in newborn babies as our evaluation could show. We conclude that the nonparametric estimation of percentiles is more adequate for the description than the use of simple regression models. PMID- 8413348 TI - [Effect of 2 different dosages of a porcine surfactant on pulmonary gas exchange of premature infants with severe respiratory distress syndrome]. AB - BACKGROUND: Doses between 20 and 200 mg/kg body weight (bw) of different surfactant preparations have been recommended in clinical trials for the treatment of neonatal RDS; an optimal dose regimen of surfactant replacement therapy has not yet been defined. Aim of the present pilot study was the evaluation of pulmonary gas exchange in infants with severe RDS following the application of either a high (200 mg/kg bw) or a low (100 mg/kg bw) dose of a natural porcine surfactant (Curosurf). METHODS: 15 neonates were randomized to a high dose regimen, 17 infants to a low dose of surfactant. Apart from a lower 1 minute Apgar in the 100 mg/kg bw group, birth weight, gestational age, sex, 5 minute-Apgar and disease severity (arterial to alveolar oxygenation ratio (a/A ratio): 0.10 +/- 0.03 [high dose], 0.11 +/- 0.06 [low dose], mean +/- SD) were well matched in both groups. RESULTS: Following surfactant instillation there was a rapid improvement in oxygenation in both groups. The a/A-ratio was slightly higher in the 200 mg/kg bw group during the first 12 hours following surfactant replacement, but statistically this was significantly higher only 4 hours after treatment (0.38 +/- 0.11 vs. 0.24 +/- 0.13, mean +/- SD, p < 0.05). CONCLUSION: The dose of 100 mg/kg bw Curosurf resulted in a rapid improvement in oxygenation and ventilatory requirements; only during the first hours following surfactant replacement there was a slight further improvement with the higher dose of 200 mg/kg bw. The impact of different dose regimens on outcome parameters still has to be defined by a larger clinical trial. PMID- 8413349 TI - [Molecular genetic diagnosis of Wiskott-Aldrich syndrome]. AB - BACKGROUND/AIMS: Wiskott-Aldrich syndrome is a severe X-linked recessive disorder of the hematopoietic system. The gene locus for Wiskott-Aldrich syndrome was mapped on the proximal short arm of the X chromosome by demonstrating close linkage to the loci DXS255 and TIMP. Carriers for Wiskott-Aldrich syndrome are asymptomatic and, hence, can not be identified clinically. METHODS: For a better estimate of the carrier risk of female family members, an extended molecular genetic analysis has been carried out on two kindreds with Wiskott-Aldrich syndrome: We followed the allele segregation at the two marker loci mentioned above known to be closely linked to the disease locus (indirect genotype diagnostics); in addition, we determined the pattern of X-inactivation by analyzing the methylation status of the two X chromosomes. RESULTS: We show that carriers for Wiskott-Aldrich syndrome can reliably be identified by the combination of segregation and X chromosome inactivation studies. Helpful information can be obtained by such studies in sporadic cases, too, or in families, in which--due to the early death--no surviving affected males are available for an DNA study. CONCLUSION: Indirect genotype analysis combined with the study of X-inactivation pattern is a valuable diagnostic tool for genetic counselling of families with Wiskott-Aldrich syndrome. PMID- 8413350 TI - [Comment on the contribution by J. Krahe et al. Secondary rhabdomyolysis and acute renal failure in gastroenteritis with hypernatremic dehydration]. PMID- 8413351 TI - [Diabetes mellitus in childhood and adolescence]. PMID- 8413353 TI - The human sensory unit and pain: new concepts, syndromes, and tests. PMID- 8413354 TI - Central pain: "new" syndromes and their evaluation. AB - Central pain syndrome is defined as pain associated with a lesion of the central nervous system. It has a low incidence but is frequently intractable and does not have effective treatment. The cause of central pain is speculative; however, the single common sensory abnormality in patients with central pain is interruption of spinothalamocortical nociceptive pathways. It appears that severe central nervous system lesions, with total destruction of ascending sensory systems, do not lead to a central pain syndrome; and that setting of mild, moderate, or severe disruption of the anterolateral ascending system with partial or complete preservation of the dorsal column/medial lemniscus functions is most frequently associated with central pain syndrome. Furthermore, even during remission, dysesthesias and pain could be triggered by additional afferent input to the large fiber/dorsal column/medial lemniscus system and, once established, they may not be abolished by additional deafferentation. PMID- 8413355 TI - Microneurography, impulse conduction, and paresthesias. AB - It is possible to learn more about peripheral nerve function in human subjects than is obtainable with routine nerve conduction studies, and thereby to study the basis of "positive" symptoms, such as paresthesias. Using microneurography, ectopic impulse activity in cutaneous afferents has been recorded in patients suffering from neurologic disorders and in normal subjects in whom paresthesias were provoked by hyperventilation, prolonged tetanization of cutaneous nerves and ischemia. Using relatively simple modifications of standard nerve conduction techniques, the increases in axonal excitability responsible for this ectopic activity have been documented in human volunteers. Hyperventilation increases axonal excitability but does not change supernormality, probably because Na+ channels are activated by the decrease in [Ca2+] on the axonal membrane. Prolonged tetanic stimulation and ischemia probably share similar mechanisms. At least in motor axons, postischemic ectopic activity occurs when the hyperpolarization that results from activation of the Na+/K+ pump lowers the membrane potential below the equilibrium potential for K+. A high extracellular [K+] can then result in an inward current producing depolarization and possibly triggering regenerative processes. PMID- 8413356 TI - Muscle pain: animal and human experimental and clinical studies. AB - The search for the identification of the sensory apparatus encoding muscle pain sensation in humans is recounted. Basic neurophysiologic animal studies, leading to a description of slowly conducting afferent from muscle and definition of high threshold polymodal muscle nociceptors, and pioneer psychophysic human studies together with recent microneurographic experiments in humans are described. The phenomena of muscle pain broad localization and distant referral are discussed, and clinical implications are extrapolated to interpret muscle pain as a localizing sign of mononeuropathy or radiculopathy. The identification of human muscle nociceptors has defined the scientific standard to test emerging clinical descriptions having muscle pain as a symptom. PMID- 8413357 TI - An animal model of neuropathic pain: a review. AB - Recent work has succeeded in producing models of painful peripheral neuropathies in laboratory animals. There is evidence that the animals experience both abnormal spontaneous pain and abnormal evoked pains (allodynia and hyperalgesia). Experimental analyses of these models have demonstrated potential pathophysiologic mechanisms in both the peripheral and central nervous systems; it is likely that the model neuropathic pain syndromes are due to several different mechanisms. One line of evidence suggests that these pain states gradually become centralized due to an excitotoxic effect on spinal cord dorsal horn inhibitory interneurons. The role of the sympathetic nervous system appears to vary, depending on the type of nerve injury and the temporal evolution of the syndrome. There is evidence indicating that the abnormality of cutaneous temperature regulation that often accompanies painful peripheral neuropathy is not necessarily due to the activity of sympathetic vasomotor efferents. PMID- 8413358 TI - Use and misuse of conventional electrodiagnosis, quantitative sensory testing, thermography, and nerve blocks in the evaluation of painful neuropathic syndromes. AB - A number of laboratory tests are critically important in the quest to diagnose presence or absence of organic neuropathic dysfunction and to establish the relevance of such to the subjective pain complaints. However, none of these tests has absolute diagnostic value and their results must be interpreted in the light of the clinical picture. Conventional electrophysiology evaluates function of large caliber afferent and motor fibers leaving the function of small caliber afferent fibers unexplored, and cannot explore the basis for positive sensory phenomena. The quantitative somatosensory thermotest is the best test available to explore function of small caliber afferents. It allows documentation of positive sensory phenomena in the form of thermal hyperalgesia. Because it is a psychophysical test, it lacks localizing value. Thermography sensitively detects and precisely delineates areas of cutaneous thermal change of neural origin. Three types of diagnostic neurologic blocks are used in the clinic: compression ischemia, local anesthetic and sympathetic blocks. Although they may provide important information about the pathophysiology of pain and hyperalgesias, adequate placebo control is of the essence because chronic neuropathic pain patients may express a high incidence of placebo response. PMID- 8413359 TI - Human microneurography and intraneural microstimulation in the study of neuropathic pain. AB - Psychophysical experiments in combination with microneurography and intraneural microstimulation in awake human subjects have yielded some useful information on somatosensory functions under normal and pathologic conditions. Normally, pain is signaled by nociceptive afferents, and tactile sensations are evoked from activation of low-threshold mechanoreceptors. Following tissue injury, nociceptors are sensitized, and their enhanced responsiveness correlates with hyperalgesia to heat and in some cases to mechanical stimuli. In addition, ongoing activity in sensitized nociceptive afferents may lead to central sensitization in such a way that normally nonpainful gentle stroking the skin evokes pain from activation of low-threshold mechanoreceptors. This particular change in signal processing in the central nervous system is restored when the ongoing nociceptive input is interrupted, whereas other forms of central sensitization can outlast the duration of the nociceptive input. PMID- 8413360 TI - The cutaneous and mixed nerve silent period. PMID- 8413361 TI - Improvement of the exercise test after therapy in thyrotoxic periodic paralysis. PMID- 8413362 TI - Progressive unilateral hypertrophy myopathy. PMID- 8413363 TI - Neurologic improvement of adrenomyeloneuropathy after steroid replacement therapy. PMID- 8413364 TI - Correlation between steroid myopathy and the elevated serum lactic dehydrogenase (LDH) 1, 2, and 3 isoenzymes. PMID- 8413365 TI - Unilateral trigeminal motor neuropathy as a presenting feature of neurofibromatosis type 2 (NF2) PMID- 8413366 TI - The potential for gene therapy in Duchenne muscular dystrophy and other genetic muscle diseases. AB - Dystrophin cDNAs have been introduced into skeletal muscle fibers of dystrophin deficient mice (mdx) through direct DNA injection in plasmid expression vectors and by replication-defective recombinant adenovirus vectors. The introduced genes appear to protect those muscle fibers from necrosis in which they become expressed. By direct injection of dystrophin cDNA in plasmid expression vector, only 1-2% of adult mdx muscle fibers of the injected muscle expressed dystrophin. On the other hand, by recombinant adenovirus injection into very young mdx muscle, a better efficiency has been reported. We have discussed several putative and proven factors that may contribute to the thus far demonstrated relatively low efficiency of dystrophin gene transfer. These include poor uptake of gene constructs by muscle fibers, degradation of the injected DNA, and poor access of gene constructs to the nuclear compartment. Neutralization or elimination of these factors could improve the efficiency of gene transfer so that it might, in the future, qualify as an effective therapy for DMD and some other genetic diseases of muscle. PMID- 8413367 TI - Magnetic stimulation in hemifacial spasm and post-facial palsy synkinesis. AB - The facial nerve was stimulated trascranially with a magnetic stimulator in 14 normal controls, 14 hemifacial spasm patients, and 16 post-facial-palsy synkinesis patients. Magnetic stimulation in normal controls revealed muscle responses which had latencies with a mean value of 4.99 +/- 0.49 ms and amplitudes of 2.41 +/- 1.08 mV. In the same group, transosseal conduction time was calculated to be 1.20 +/- 0.13 ms. In the hemifacial spasm group, the amplitudes of the responses on the affected sides were lower as compared to the unaffected sides (mean values 1.78 vs. 2.41 mV, P = 0.01). Also, the threshold to magnetic stimulation was elevated on the affected sides. These findings are suggestive of the presence of a hypoexcitability to magnetic stimulation in the root entry zone. In the post-facial-palsy synkinesis patients, magnetic stimulation of the affected sides resulted in responses with long latencies and low amplitudes (mean latency 6.34 ms, mean amplitude 0.90 mV). In the recordings made with magnetic stimulation, the difference of the latencies between the two sides was larger as compared to those obtained by electrical stimulation. The transosseal conduction time was also remarkably prolonged on the affected side. These findings may suggest that magnetic stimulation can be an effective method of showing intracranially located lesions of the facial nerve. PMID- 8413368 TI - Molecular analysis of the Duchenne muscular dystrophy gene in patients with Becker muscular dystrophy presenting with dilated cardiomyopathy. AB - Molecular analysis of the Duchenne muscular dystrophy (DMD) gene was performed on 4 unrelated patients with Becker muscular dystrophy (BMD) presenting with dilated cardiomyopathy. Two patients with a deletion involving exon 1 were quite unique in that they developed fatal myocardial involvement in their teens, despite the absence of significant muscular weakness. The deletion found in these patients comprised the 3'-end of exon 1 and the greater part of intron 1. Two other patients with a deletion of exon 47 showed progressive muscular atrophy and weakness; they were considered to be typical BMD in both clinical features and the type of gene deletion. We speculate that a deletion around exon 1 may severely damage the expression and/or the function of dystrophin selectively in cardiac muscle, but not in skeletal muscle. PMID- 8413369 TI - Pediatric peroneal mononeuropathy: a clinical and electromyographic study. AB - Seventeen children with pediatric peroneal mononeuropathies evaluated between 1979 and 1991 are reported. Twelve boys and 5 girls, ranging in age from 1.5 months to 17 years, were referred for footdrop in 16 children (94%) or for lower extremity pain in 1 child (6%). Causes included compression in 10 children (59%), trauma in 3 children (18%), entrapment in 3 children (18%), and indeterminate in 1 child (5%). Based on nerve conduction studies and electromyography, the level of the pediatric peroneal mononeuropathic lesion was the common peroneal nerve in 10 children (59%), the deep peroneal nerve in 2 children (12%), and the superficial peroneal nerve in 1 child (5%). In 4 other children (24%), pediatric peroneal mononeuropathy at the knee was not more precisely identified. Surgical exploration in 3 children with progressive pediatric peroneal mononeuropathy was valuable. Improvement occurred in 13 of 17 children (76%). PMID- 8413370 TI - CMAP dispersion, amplitude decay, and area decay in a normal population. AB - The aim of this investigation was to define the boundaries of compound motor action potential (CMAP) dispersion, amplitude decay, and area decay in a control population and determine their dependence on external variables such as age and interelectrode distance. Measurements were made from median, ulnar, and common peroneal motor nerves of 110 normal subjects of ages 15-90 years. Significant differences between nerves were found in mean values of each parameter. Dispersion and amplitude decay increased with the square of age in all three nerves, while area decay increased with age in the median nerve. Dispersion was the main cause of amplitude decay. Use of regression equations to predict dispersion and amplitude decay in each nerve significantly tightened confidence limits and should therefore increase the accuracy of these parameters in detecting demyelinating pathology in peripheral nerve. PMID- 8413371 TI - Selective deep peroneal nerve injury associated with arthroscopic knee surgery. AB - We report 3 cases of isolated deep peroneal nerve injury as a complication of arthroscopic knee surgery. At the level of the knee joint, the deep and superficial peroneal nerves are usually joined as the common peroneal nerve. However, because of the fascicular structure, a partial nerve injury can result in an isolated injury to the deep peroneal nerve fibers. Due to the intraneural topography of the peroneal nerve, electrodiagnostic studies in a partial nerve injury may erroneously indicate a more distal lesion. PMID- 8413372 TI - Peripheral neuromuscular manifestations in systemic sclerosis (scleroderma). AB - Systemic sclerosis (scleroderma) is thought to be the least likely of the collagen vascular disorders to cause nervous system damage. We evaluated the peripheral neuromuscular manifestations in 32 patients with scleroderma. A clinically defined peripheral nervous system (PNS) lesion was manifest in 5 of 32 patients (16%), including 2 patients with trigeminal neuropathy and single cases of polyneuropathy, brachial plexopathy, and lumbosacral radiculopathy. Neurophysiological studies suggested subclinical PNS involvement in 6 additional patients (3 with distal axonal polyneuropathy, 1 with probable myopathy and superimposed polyneuropathy, 1 with trigeminal neuropathy, and 1 with focal ulnar neuropathy). Even though subjective muscular complaints were numerous (16 patients, 50%), a defined primary muscular disease could be demonstrated only in 5 patients (16%). Our results indicate that peripheral neuropathy in scleroderma is not as uncommon as previously estimated. PMID- 8413373 TI - Motor unit number estimation, isometric strength, and electromyographic measures in amyotrophic lateral sclerosis. AB - Pathologic progression in amyotrophic lateral sclerosis (ALS) results from motor neuron death, while the clinical expression also reflects the compensatory effects of collateral reinnervation consequent to lower motor neuron loss. In a cross-sectional study of ALS subjects, we made comparisons between motor unit number estimation (MUNE) values and several measures reflecting collateral reinnervation, including isometric strength, compound muscle action potential (CMAP) amplitude, surface motor unit action potential (S-MUAP) amplitude, fiber density (FD), macro-EMG potential amplitude, turns-to-amplitude (T/A) ratio, and amplitude and recruitment pattern of low threshold voluntary motor units in elbow flexor muscles. Before comparisons were made, test-retest reproducibility of these measures was assessed in ALS subjects, and is highest for isometric strength, and lower but similar for EMG measures. When the effects of multiple comparisons are considered, borderline significant correlations are found between MUNE values and isometric strength. Neither MUNE values nor isometric strength are significantly correlated with macro-EMG amplitude, FD, T/A ratio, or amplitude and recruitment rate of low threshold voluntary motor units. There are significant correlations of CMAP and S-MUAP with MUNE values, but these are statistical artifacts with no independent interpretation. We conclude that collateral reinnervation prevents isometric strength and EMG measures from accurately reflecting lower motor neuron death in ALS. MUNE measurements are better suited to provide insight into the true natural history of the disease process and may be clinically useful to follow progression and response in drug trials. PMID- 8413374 TI - Somatosensory evoked potentials induced by stimulating a variable number of nerve fibers in rat. AB - Somatosensory evoked potentials (SEPs) were recorded from rat spinal cord (sSEPs) and cerebral cortex (cSEPs). Stimulus sites included either one or both sural nerve branches having different fiber populations (group A), or distal to a lesion of controlled size of the sural nerve made 1 week earlier (group B). In the two groups of animals, amplitudes of SEPs correlated with the quantity of large myelinated nerve fibers. Peak latencies of sSEPs in group A related to the ratio of sizes of transmitting fibers. sSEPs and cSEPs in both groups A and B could be recorded in a reproducible fashion by stimulating sural nerve branches or lesioned nerve trunks containing only 100 or less nerve fibers greater than 4 microns in size. Thus, presence of sSEPs or cSEPs after stimulation distal to a lesion site does not insure that many nerve fibers have continuity with the central nervous system (CNS). PMID- 8413375 TI - Muscle cramp as the result of impaired GABA function--an electrophysiological and pharmacological observation. AB - We investigated the mechanism of cramps in 2 patients: a 48-year-old man with bulbospinal neuronopathy, and a 46-year-old man with amyotrophic lateral sclerosis. Cramps were quite easily induced by volitional exertion and high frequency stimulation of the peripheral nerves. When an ulnar nerve was blocked with lidocaine at the elbow, no cramp was induced despite the application of high frequency stimulation at the wrist. Diazepam (GABAA agonist) was effective in the first patient and baclofen (GABAB agonist) in the second, with no cramps induced in spite of increasing stimulation intensity. Impairment of interneurons mediated by GABA as the neurotransmitter is thought to be involved in the mechanism of the cramps. PMID- 8413376 TI - Screening of dominantly inherited Charcot-Marie-Tooth neuropathies. AB - Sixty-three families with dominantly inherited Charcot-Marie-Tooth (CMT) neuropathies including 730 subjects (total) from which 356 affected were studied clinically, electrophysiologically (MNCVs and EMGs), by genetic linkage, and screened for DNA duplication. Thirty-eight families (60.3%) were type 1A (demyelinating CMT mapped on chromosome 17). DNA duplication was present in 36 families (94.8% of CMT1A families). One CMT1A family (2.6%) showed no duplication but suggested genetic linkage with markers of chromosome 17. One CMT1A family (2.6%) revealed nonduplication in some affected members and duplication in other affected members. The disease in that family segregated with the same chromosome 17 markers regardless of duplication status. The other CMT families with dominant inheritance but without duplication included one family with CMT1B (demyelinating CMT mapped on chromosome 1) (1.6%), 14 families with CMT2 axonal neuropathy (22.2%), and 10 families with X-linked dominant CMT (15.9%). PMID- 8413377 TI - Development and reversal of fatigue in human tibialis anterior. AB - The lack of easily measurable, objective physiologic indices of muscle fatigue in humans has hampered the evaluation of interventions that putatively inhibit muscle fatigue. In 6 healthy subjects, isometric fatigue of right and left tibialis anterior (TA) was produced using 180 s of intermittent electrical stimulation to the motor point (40 Hz, 650-ms or 250-ms trains, 1 train/s). The time course, magnitude, and reproducibility of fatigue and recovery from fatigue was established, and the effect of duty cycle measured. During fatigue, force fell rapidly in all subjects over the first 70 s, then fell more slowly. Recovery was also biphasic--force increased rapidly over the initial 30 s then slowly--and was incomplete at 300 s. Comparing duty cycles of 0.65 versus 0.25, the longer duty cycle caused a greater fall in force (-65% versus -29%; P < 0.001) and lower recovery (P < 0.02). In all subjects, left leg muscles fatigued more than right. We conclude that this noninvasive fatigue and recovery protocol is highly reproducible and that it may prove useful for evaluating the effects of pharmacologic or restorative interventions on human muscle fatigue. PMID- 8413378 TI - Histochemical and contractile property changes during human muscle development. AB - When the histochemical and contractile properties of infant muscles change postnatally, and what influence muscle function has on these changes, were the focus of this study. Contractile properties were measured in the plantaflexor (PF) and dorsiflexors (DF) of 19 newborns and 36 infants aged 5-16 months. Infants were tested between one and four times at monthly intervals. Measurements included maximal twitch tension (Pt), time to peak tension (TPT), and half relaxation times (1/2RT). TPT was similar in PF (77 ms) and DF (73 ms) at birth, remained unchanged in DF, and slowed in PF to 110-120 ms between 9 and 12 months. Type I distributions were determined at autopsy in fetal through to adult muscles. Completion of differentiation occurred in soleus at about the age that contractile properties slowed and infants started to use these muscles more. A trend of higher percentages of type I distributions was also noted in children than either newborns or adults in other muscles. The implications of these findings and the clinical use of these methods for evaluating peripheral neuromuscular function is discussed. PMID- 8413379 TI - Orthostatic tremor: clinical and electrophysiologic characteristics. AB - Orthostatic tremor, sometimes known as "shaky legs syndrome," is a disorder of middle-aged or elderly people characterized by feelings of unsteadiness in the legs and a fear of falling when standing. Patients stand on a wide base but walk normally. These symptoms are due to high-frequency (13-18 Hz) burst firing in weight-bearing muscles. They are attenuated by walking and are abolished immediately by sitting. Some authors believe that the disorder is a variant of essential tremor. This study reports the clinical and electrophysiologic features of orthostatic tremor in 30 patients. The findings indicate that orthostatic tremor is distinct from essential tremor, both clinically and electrophysiologically. The major differences are the frequency of electromyographic burst firing, the invariable involvement of lower limb and paraspinal muscles, and the task-specific nature of the tremor in orthostatic tremor. The study shows that the diagnosis can be established rapidly with surface electromyographic recordings. PMID- 8413380 TI - Acute Lyme neuropathy presenting with polyradicular pain, abdominal protrusion, and cranial neuropathy. AB - A 53-year-old man developed multifocal radicular pain. The diagnosis of Lyme neuroborreliosis was delayed until bifacial paresis and right lower abdominal wall weakness developed, prompting further evaluation. Cerebrospinal fluid (CSF) examination showed aseptic meningitis. Antibodies directed against Borrelia burgdorferi were present in the serum; higher titers were present in the CSF, indicating local antibody production. Electrophysiologic studies showed both an axonal polyradiculopathy as well as demyelinative facial palsy. Ceftriaxone therapy led to marked improvement in pain and facial palsies. PMID- 8413381 TI - Sensitivity of electrophysiological studies and the carpal tunnel syndrome. PMID- 8413382 TI - An indirect immunofluorescence test for the detection of acetylcholine receptor antibodies. PMID- 8413383 TI - Beneficial effect of plasmapheresis on Fisher's syndrome. PMID- 8413384 TI - Motor evoked potentials in orbicularis oris muscle: no evidence of ipsilateral corticonuclear projections. PMID- 8413385 TI - Acute alcoholic rhabdomyolysis associated with abnormal ischemic exercise test. PMID- 8413386 TI - Diabetic proximal neuropathy. PMID- 8413387 TI - Prevention of preeclampsia with low-dose aspirin in healthy, nulliparous pregnant women. The National Institute of Child Health and Human Development Network of Maternal-Fetal Medicine Units. AB - BACKGROUND: Although low-dose aspirin has been reported to reduce the incidence of preeclampsia among women at high risk for this complication, its efficacy and safety in healthy, nulliparous pregnant women are not known. METHODS: We studied 3135 normotensive nulliparous women who were 13 to 26 weeks pregnant to determine whether treatment with aspirin reduced the incidence of preeclampsia. Of this group, 1570 women received 60 mg of aspirin per day and 1565 received placebo for the remainder of their pregnancies. We also evaluated the effect of aspirin on maternal and neonatal morbidity. RESULTS: Of the original group of 3135 women, 2985 (95 percent) were followed throughout pregnancy and the immediate puerperium. The incidence of preeclampsia was lower in the aspirin group (69 of 1485 women [4.6 percent]) than in the placebo group (94 of 1500 women [6.3 percent]) (relative risk, 0.7; 95 percent confidence interval, 0.6 to 1.0; P = 0.05), whereas the incidence of gestational hypertension was 6.7 and 5.9 percent, respectively. There were no significant differences in the infants' birth weight or in the incidence of fetal growth retardation, postpartum hemorrhage, or neonatal bleeding problems between the two groups. Subgroup analysis showed that preeclampsia occurred primarily in women whose initial systolic blood pressure was 120 to 134 mm Hg (incidence among such women, 5.6 percent in the aspirin group vs. 11.9 percent in the placebo group; P = 0.01). The incidence of abruptio placentae was greater among the women who received aspirin (11 women, vs. 2 in the placebo group; P = 0.01). CONCLUSIONS: Low-dose aspirin decreases the incidence of preeclampsia among nulliparous women, primarily through its effect in those who have elevated systolic blood pressure initially. This treatment does not decrease perinatal morbidity but increases the risk of abruptio placentae. PMID- 8413388 TI - Cyclosporine, methotrexate, and prednisone compared with cyclosporine and prednisone for prophylaxis of acute graft-versus-host disease. AB - BACKGROUND: Acute graft-versus-host disease (GVHD) following allogeneic bone marrow transplantation remains a serious problem. In a clinical trial, we tested the combination of cyclosporine and prednisone with and without methotrexate for the prevention of GVHD. METHODS: One hundred fifty patients with either acute leukemia in first complete remission, chronic myelogenous leukemia in first chronic phase, or lymphoblastic lymphoma in first complete remission were enrolled in the study. All the patients were given fractionated total-body irradiation (1320 cGy) and etoposide (60 mg per kilogram of body weight) in preparation for transplantation, and received bone marrow from genotypically histocompatible donors. To prevent GVHD, they were randomly assigned to prophylactic treatment with either cyclosporine, methotrexate, and prednisone or cyclosporine and prednisone without methotrexate. All the patients received standardized supportive care after transplantation, including intravenous gamma globulin. RESULTS: Patients receiving cyclosporine, methotrexate, and prednisone had a significantly lower incidence of acute GVHD of grades II to IV (9 percent) than those receiving cyclosporine and prednisone (23 percent, P = 0.02). Multivariate regression analysis demonstrated that an increased risk of acute GVHD was associated with an elevated serum creatinine concentration (P = 0.006) and treatment with cyclosporine and prednisone alone (P = 0.02). The lower incidence of acute GVHD was not associated with a higher rate of relapse of leukemia or lymphoma. There was no significant difference in disease-free survival at three years between the two treatment groups (64 percent with the three-drug regimen vs. 59 percent with the two-drug regimen, P = 0.57). CONCLUSIONS: The combination of cyclosporine, methotrexate, and prednisone was more effective in preventing acute GVHD of grades II to IV than was the combination of cyclosporine and prednisone without methotrexate. PMID- 8413389 TI - Overwhelming pulmonary blastomycosis associated with the adult respiratory distress syndrome. AB - BACKGROUND AND METHODS: Few cases of overwhelming pulmonary blastomycosis associated with the adult respiratory distress syndrome have been reported. We describe 10 patients with this condition who were treated at one center in Wisconsin. RESULTS: All 10 patients presented with fever, cough, and dyspnea; radiographic evidence of diffuse pulmonary infiltrates; and marked impairment of oxygenation. The mean alveolar-arterial oxygen gradient was 616 mm Hg. Six of the patients had no underlying disease associated with altered immunity, and two had no recent exposure to environmental reservoirs of Blastomyces dermatitidis. In all 10 patients, large numbers of broad-based budding yeasts were seen on microscopical examination of tracheal secretions. All patients were treated with intravenous amphotericin B (0.7 to 1.0 mg per kilogram per day). Of the five survivors, four received full doses of amphotericin B in the first 24 hours, and four required mechanical ventilatory support for 7 to 151 days. Long-term follow up of three survivors showed good recovery of pulmonary function. CONCLUSIONS: Overwhelming infection with B. dermatitidis can cause diffuse pneumonitis and the adult respiratory distress syndrome, even in immunocompetent hosts. With prompt diagnosis by microscopical examination of tracheal secretions, intensive therapy with amphotericin B, and ventilatory support, good recovery of pulmonary function is possible. PMID- 8413390 TI - Brief report: occult pseudoxanthoma elasticum in patients with premature cardiovascular disease. PMID- 8413391 TI - Images in clinical medicine. Pseudoxanthoma elasticum. PMID- 8413392 TI - The methodologic foundations of studies of the appropriateness of medical care. PMID- 8413393 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 42-1993. Jaundice and anemia two weeks after an aortic valvuloplasty in a 62-year-old woman with splenomegaly. PMID- 8413394 TI - The quality of care and the quality of measuring it. PMID- 8413395 TI - Preventing preeclampsia. PMID- 8413396 TI - Progress in radioimmunotherapy. PMID- 8413397 TI - Changing physicians' practices. PMID- 8413398 TI - Abdominal aortic aneurysm. PMID- 8413399 TI - Abdominal aortic aneurysm. PMID- 8413400 TI - Abdominal aortic aneurysm. PMID- 8413401 TI - Transcutaneous pacing in patients with asystolic cardiac arrest. PMID- 8413402 TI - Outcomes after heart-valve replacement. PMID- 8413403 TI - Recent advances in pulmonary medicine. PMID- 8413404 TI - Recent advances in pulmonary medicine. PMID- 8413405 TI - Fever in patients with neutropenia. PMID- 8413406 TI - Fever in patients with neutropenia. PMID- 8413407 TI - Fever in patients with neutropenia. PMID- 8413408 TI - Competitiveness, peer pressure, and career choice. PMID- 8413409 TI - Treatment of acute lymphoblastic leukemia. 30 years' experience at St. Jude Children's Research Hospital. AB - BACKGROUND: Therapy for childhood lymphoblastic leukemia has evolved during the past three decades, but key questions about what are the least toxic, most effective forms of treatment remain unanswered because of the lack of comprehensive follow-up information. METHODS: To assess long-term outcome in the series of clinical trials conducted at St. Jude Hospital, we compared the results of treatment typical of four eras: exploratory combination chemotherapy (era 1, 1962 to 1966; 91 patients), regimens for the control of meningeal leukemia (era 2, 1967 to 1979; 825 patients), limited intensification of therapy (era 3, 1979 to 1983; 428 patients), and extended intensification of therapy (era 4, 1984 to 1988; 358 patients). ("Intensification" refers to strategies of systemic chemotherapy that are more aggressive than conventional ones.) The major end points were survival and event-free survival; we also calculated the relative risk of treatment failure and the rate of relapse or death after treatment ended (post-treatment failure rate). RESULTS: The probability of event-free survival improved significantly in each successive era (P < 0.001 by the log-rank test), reaching 71 percent in era 4. There was a decrease of approximately 50 percent in the risk of treatment failure from one era to the next in each subgroup of patients defined according to different combinations of the leukocyte count, race, age, and sex. Leukemia appeared to be eradicated in patients who remained in complete remission for three years or more after treatment in era 4. The incidence of death due to nonleukemic causes remained 4 to 6 percent despite the trend toward more intensive treatment. An estimated 765 patients (45 percent) are long-term survivors; most of them (80 percent) have no health problems related to leukemia or its treatment. CONCLUSIONS: The development and successful application of preventive therapy for meningeal leukemia, followed by the intensification of systemic chemotherapy, has progressively improved the rate of cure of childhood lymphoblastic leukemia, with relatively few adverse sequelae. PMID- 8413410 TI - Pretransplantation burden of leukemic progenitor cells as a predictor of relapse after bone marrow transplantation for acute lymphoblastic leukemia. AB - BACKGROUND: We developed a test to discern small numbers of residual leukemic progenitor cells in the bone marrow of patients with acute lymphoblastic leukemia (ALL) in remission. Preliminary studies revealed that before undergoing bone marrow transplantation such patients differed in their burden of leukemic progenitor cells. These observations suggested that the burden of these cells might influence the risk of relapse after transplantation. METHODS: The number of residual leukemic progenitor cells before bone marrow transplantation was determined for 83 patients with high-risk ALL. We combined multiparameter flow cytometry and cell sorting with assays for leukemic progenitor cells in a quantitative method for the detection of minimal residual disease. RESULTS: The count of leukemic progenitor cells in bone marrow specimens from patients in remission varied markedly between patients, ranging from 0 to 12,546 cells per million mononuclear cells, or from 0 to 1.255 percent (median, 51 leukemic progenitor cells per million mononuclear cells, or 0.005 percent). Patients whose count of leukemic progenitor cells exceeded the median value had a higher likelihood of relapse than did patients with values below the median (relapse rate at one year, 100 percent vs. 41 percent; P < 0.001). There was a statistically significant inverse relation between the leukemic progenitor-cell content of bone marrow before transplantation and the duration of remission after transplantation (P < 0.001). The estimated risk of relapse for patients with > or = 51 leukemic progenitor cells per million mononuclear cells was more than 3.5 times the risk for patients with lower counts, after adjustment for the effects of other covariates (P = 0.005). CONCLUSIONS: The count of residual leukemic progenitor cells is a powerful predictor of relapse after autologous bone marrow transplantation, particularly among male patients. Its measurement may be useful for analyzing and improving the treatment of patients with high-risk ALL in remission. PMID- 8413411 TI - Brief report: non-corticosteroid-associated osteonecrosis of the femoral heads in two patients with inflammatory bowel disease. PMID- 8413412 TI - Images in clinical medicine. Sting of the fire ant (Solenopsis). PMID- 8413413 TI - Clinical implications of the p53 tumor-suppressor gene. PMID- 8413414 TI - Management of urinary tract infections in adults. PMID- 8413415 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 43-1993. A 71-year-old woman with confusion, hemianopia, and an occipital mass. PMID- 8413416 TI - Acute lymphoblastic leukemia--progress in children, less in adults. PMID- 8413417 TI - Participation of physicians in capital punishment. PMID- 8413418 TI - Screening for colorectal cancer. PMID- 8413419 TI - Screening for colorectal cancer. PMID- 8413420 TI - Screening for colorectal cancer. PMID- 8413421 TI - Screening for colorectal cancer. PMID- 8413422 TI - Screening for colorectal cancer. PMID- 8413423 TI - Screening for colorectal cancer. PMID- 8413424 TI - Blood transfusions and prognosis in colorectal cancer. PMID- 8413425 TI - Blood transfusions and prognosis in colorectal cancer. PMID- 8413426 TI - More on the eradication of Helicobacter pylori and the recurrence of duodenal ulcer. PMID- 8413427 TI - Antiemetic effects of ondansetron and metopimazine. PMID- 8413428 TI - Relapse of hyperprolactinemia revisited. PMID- 8413429 TI - The American Health Security Act. PMID- 8413430 TI - The American Health Security Act. PMID- 8413431 TI - A comparison of real-time compression ultrasonography with impedance plethysmography for the diagnosis of deep-vein thrombosis in symptomatic outpatients. AB - BACKGROUND: Impedance plethysmography performed serially over a one-week period has been shown to be an effective diagnostic strategy for patients with clinically suspected acute deep-vein thrombosis. Compression ultrasonography has a high sensitivity and specificity for the detection of proximal-vein thrombosis. The clinical value of repeated ultrasonography in the management of symptomatic deep-vein thrombosis is unknown. METHODS: We conducted a randomized trial in 985 consecutive outpatients with clinically suspected deep-vein thrombosis to compare the diagnostic value of serial impedance plethysmography (494 patients) and serial compression ultrasonography (491 patients). We compared the positive predictive values of both tests for the diagnosis of venous thrombosis, using contrast venography as a reference. The frequencies of venous thromboembolism during a six-month follow-up period were also compared in patients with repeatedly normal results in order to evaluate the safety of withholding anticoagulant therapy from such patients. RESULTS: The positive predictive value of an abnormal ultrasonogram was 94 percent (95 percent confidence interval, 87 to 98 percent), whereas the predictive value of impedance plethysmography was 83 percent (95 percent confidence interval, 75 to 90 percent) (P = 0.02). In patients with repeatedly normal results, the incidence of venous thromboembolism during the six-month follow-up period was 1.5 percent (95 percent confidence interval, 0.5 to 3.3 percent) for serial compression ultrasonography, as compared with 2.5 percent (95 percent confidence interval, 1.2 to 4.6 percent) for serial impedance plethysmography. CONCLUSIONS: In making the diagnosis of deep-vein thrombosis in symptomatic outpatients, serial compression ultrasonography is preferable to impedance plethysmography, in view of its superior performance in detecting venous thrombosis. PMID- 8413432 TI - A comparison of subcutaneous low-molecular-weight heparin with warfarin sodium for prophylaxis against deep-vein thrombosis after hip or knee implantation. AB - BACKGROUND: Deep-vein thrombosis is a potentially life-threatening complication of total hip or knee replacement. There are few data on the effectiveness and safety of warfarin as compared with low-molecular-weight heparin as prophylaxis against this problem. METHODS: We therefore performed a randomized, double-blind trial in 1436 patients to evaluate the effectiveness and safety of low-molecular weight heparin (given subcutaneously once daily) as compared with adjusted-dose warfarin to prevent venous thrombosis after hip or knee replacement. Treatment with the drugs was started postoperatively. The primary end point was deep-vein thrombosis as detected by contrast venography (performed a mean of 9.4 days after surgery in each group). RESULTS: Among the 1207 patients with interpretable venograms, 231 of 617 patients (37.4 percent) in the warfarin group and 185 of 590 patients (31.4 percent) in the low-molecular-weight-heparin group had deep vein thrombosis (P = 0.03). The reduction in risk with low-molecular-weight heparin as compared with warfarin was 16 percent, and the absolute difference in the incidence of venous thrombosis was 6 percent in favor of low-molecular-weight heparin (95 percent confidence interval, 0.8 to 11.4 percent). The incidence of major bleeding was 1.2 percent (9 of 721 patients) in the warfarin group and 2.8 percent (20 of 715 patients) in the low-molecular-weight-heparin group (P = 0.04), and the absolute difference was 1.5 percent in favor of warfarin (95 percent confidence interval, 0.1 to 3.0 percent). CONCLUSIONS: Our data demonstrate that the small reduction in the incidence of venous thrombosis with low-molecular-weight heparin, as compared with warfarin, was offset by an increase in bleeding complications. Although the use of low-molecular-weight heparin is simpler, because it is administered subcutaneously without the need for monitoring, it may be more costly than warfarin. Warfarin is inexpensive, but the overall cost of its use is increased by the need to monitor the intensity of anticoagulation. At this time it is unclear which of these approaches is the most cost effective. PMID- 8413433 TI - Selective bladder preservation by combination treatment of invasive bladder cancer. AB - BACKGROUND: For patients with invasive bladder cancer the usual recommended treatment is radical cystectomy, although transurethral resection of the tumor, systemic chemotherapy, and radiotherapy are each effective in some patients. We sought to determine whether these treatments in combination might be as effective as radical cystectomy and thus might allow the bladder to be preserved and the cancer cured. METHODS: We enrolled 53 consecutive patients with muscle-invading bladder cancer (stages T2 through T4, NXM0) in a trial of transurethral surgery, combination chemotherapy, and irradiation (4000 cGy) with concurrent cisplatin administration. Urologic evaluation of the tumor response directed further therapy: radical cystectomy in the 8 patients who had incomplete responses, additional chemotherapy and radiotherapy (6480 cGy) in the 34 patients who had complete responses or who were unsuited for cystectomy, and alternative care in the 11 patients who could not tolerate either irradiation or chemotherapy. RESULTS: After a median follow-up of 48 months, 24 of the 53 patients (45 percent) were alive and free of detectable tumor. In 31 patients (58 percent) the bladder was free of invasive tumor and functioning well, even though in 9 (17 percent) a superficial tumor recurred and required further transurethral surgery and intravesical drug therapy. Of the 28 patients who had complete responses after initial treatment, 89 percent had functioning tumor-free bladders. CONCLUSIONS: Conservative combination treatment may be an acceptable alternative to immediate cystectomy in selected patients with bladder cancer, although a randomized clinical trial that included a group for simultaneous comparison would be required to produce definitive results. PMID- 8413434 TI - Polycystic ovaries and hyperandrogenism in women taking valproate for epilepsy. AB - BACKGROUND: Reproductive endocrine disorders are more common among women with epilepsy than among normal women. These disorders have been attributed to epilepsy itself, but could be related to antiepileptic-drug therapy. METHODS: We studied 238 women with epilepsy who were seen regularly at the Outpatient Department of the University Hospital, Oulu, Finland. Their mean age was 33 years (range, 18 to 45), and the mean duration of therapy was 9 years (range, 0 to 31). Twenty-nine (12 percent) were treated with valproate, 120 (50 percent) with carbamazepine, 12 (5 percent) with valproate and carbamazepine, and 62 (26 percent) with other medications; 15 (6 percent) were untreated. Vaginal ultrasonography was performed to determine ovarian size, and serum sex-hormone concentrations were measured in 41 women with epilepsy and menstrual disturbances, 57 women with epilepsy and regular menstrual cycles, and 51 normal women. RESULTS: Menstrual disturbances were present in 13 of the women receiving valproate alone (45 percent), 3 of the women receiving valproate in combination with carbamazepine (25 percent), 23 of the women receiving carbamazepine (19 percent), and 8 of those receiving other medications (13 percent). Forty-three percent of the women receiving valproate had polycystic ovaries, and 17 percent had elevated serum testosterone concentrations without polycystic ovaries; 50 percent of the women receiving valproate and carbamazepine had polycystic ovaries, and 38 percent had elevated serum testosterone concentrations without polycystic ovaries. Eighty percent of the women treated with valproate before the age of 20 years had polycystic ovaries of hyperandrogenism. CONCLUSIONS: Menstrual disturbances, polycystic ovaries, and hyperandrogenism are often encountered in women taking valproate for epilepsy. PMID- 8413435 TI - Brief report: heavy-chain deposition disease. PMID- 8413436 TI - Images in clinical medicine. Sarcoidosis. PMID- 8413437 TI - The organization of medical care. Lessons from the Medicare end stage renal disease program. PMID- 8413438 TI - Vaccines against human immunodeficiency virus--progress and prospects. PMID- 8413439 TI - Clinical problem-solving. The heart of the matter. PMID- 8413440 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 44-1993. A 39-year-old man with fever, polyarthralgia, and purpuric skin lesions. PMID- 8413441 TI - The diagnosis and treatment of deep-vein thrombosis. PMID- 8413442 TI - New approaches to the treatment of bladder cancer. PMID- 8413443 TI - Ominous consequences of immunoglobulin deposition. PMID- 8413444 TI - Vitamin E and the risk of coronary disease. PMID- 8413445 TI - Vitamin E and the risk of coronary disease. PMID- 8413446 TI - Self-administration of Salmonella endotoxin. PMID- 8413447 TI - Self-administration of Salmonella endotoxin. PMID- 8413448 TI - More on pneumococcal appendicitis. PMID- 8413449 TI - Sleep-disordered breathing. PMID- 8413450 TI - Sleep-disordered breathing. PMID- 8413451 TI - Sleep-disordered breathing. PMID- 8413452 TI - Proximal-muscle weakness induced by herbs. PMID- 8413453 TI - Outpatient management without antibiotics of fever in selected infants. AB - BACKGROUND: In many academic centers it is standard practice to hospitalize all febrile infants younger than two months of age, whereas in community settings such infants are often cared for as outpatients. METHODS: We conducted a controlled study of 747 consecutive infants 29 through 56 days of age who had temperatures of at least 38.2 degrees C. After a complete history taking, physical examination, and sepsis workup, the 460 infants with laboratory or clinical findings suggestive of serious bacterial illness were hospitalized and treated with antibiotics. The screening criteria for serious bacterial illness included a white-cell count of at least 15,000 per cubic millimeter, a spun urine specimen that had 10 or more white cells per high-power field or that was positive on bright-field microscopy, cerebrospinal fluid with a white-cell count of 8 or more per cubic millimeter or a positive Gram's stain, or a chest film showing an infiltrate. The 287 infants who had unremarkable examinations and normal laboratory results were assigned to either inpatient observation without antibiotics (n = 148) or outpatient care without antibiotics but with reexaminations after 24 and 48 hours (n = 139). RESULTS: Serious bacterial illness was diagnosed in 65 infants (8.7 percent). Of these 65 infants, 64 were identified by our screening criteria for inpatient care and antibiotic treatment (sensitivity = 98 percent; 95 percent confidence interval, 92 to 100). Of the 287 infants assigned to observation and no antibiotics, 286 (99.7 percent) did not have serious bacterial illness. Only two infants assigned to outpatient observation were subsequently admitted to the hospital; neither was found to have a serious illness. Outpatient care without antibiotics of the febrile infants at low risk for serious illness resulted in a savings of about $3,100 per patient. CONCLUSIONS: With the use of strict screening criteria, a substantial number of febrile one-to-two-month-old infants can be cared for safely as outpatients and without antibiotics. PMID- 8413454 TI - Age-related increase in mortality among patients with first myocardial infarctions treated with thrombolysis. The Investigators of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardico (GISSI-2). AB - BACKGROUND: The overall rate of mortality due to ischemic heart disease is known to increase progressively with age. We evaluated the relation between the mortality rate and age in patients with first myocardial infarctions treated with thrombolytic therapy. METHODS: We studied 9720 patients with first infarctions who had been enrolled in the GISSI-2 trial. (This trial compared the efficacy of tissue plasminogen activator with that of streptokinase in patients with myocardial infarction.) Of these, only 35 percent had a history of angina. The relation between age and mortality during hospitalization and during the six months after discharge was determined by unadjusted and adjusted analyses. RESULTS: The in-hospital mortality rate was 1.9 percent among patients 40 years old or younger, but it increased to 31.9 percent among those more than 80 years old; however, values for indicators of infarct size did not increase with age. Autopsies were performed in 20 percent of the 772 patients who died in the hospital; the findings showed that the frequency of cardiac rupture increased from 19 percent among patients 60 years old or younger to 86 percent among those more than 70 years old. The mortality rate for the first six months after hospital discharge also increased significantly with age. After adjustment for confounding variables, older age continued to be significantly associated with a higher risk of in-hospital and post-discharge death. When age was introduced into a multivariate model as a continuous variable, the risk of death was estimated to increase by about 6 percent per year for both in-hospital and six-month mortality rates. CONCLUSIONS: In patients with first myocardial infarctions who received thrombolytic therapy, age was a powerful independent predictor of both in hospital and post-discharge mortality rates. The exponential, age-related increase in the mortality rate did not appear to be explained by larger infarcts. PMID- 8413455 TI - The diagnosis and treatment of baroreflex failure. AB - BACKGROUND: Baroreflexes originate in the great vessels of the neck and thorax and prevent arterial pressure from rising or falling excessively. METHODS: This study was undertaken to clarify the cause, clinical spectrum, and therapy of this disorder. We studied 11 patients with baroreflex failure presenting as severe, labile hypertension and hypotension, often with headache, diaphoresis, and emotional instability, and characterized by the failure of exogenous vasoactive substances to alter heart rate. Each underwent hemodynamic monitoring and biochemical, physiologic, and pharmacologic testing. RESULTS: The patients' maximal systolic blood pressures ranged from 164 to 280 mm Hg, and their minimal systolic pressures ranged from 58 to 96 mm Hg. Plasma norepinephrine and epinephrine concentrations were sometimes many times normal during blood-pressure surges. All the patients had excessive pressor and tachycardia responses to the mental-arithmetic and cold pressor tests and marked hypersensitivity to clonidine. The underlying causes of baroreflex failure included the familial paraganglioma syndrome, neck surgery or radiation therapy for pharyngeal carcinoma, bilateral lesions of the nucleus tractus solitarii, and surgical section of the glossopharyngeal nerves; in two patients the cause was unknown. Therapy with clonidine reduced the frequency of attacks by 81 percent and attenuated the elevated blood pressure and heart rate in the attacks that occurred. CONCLUSIONS: The syndrome of baroreflex failure should be considered in patients with otherwise unexplained labile hypertension. Clonidine attenuates the pressor and tachycardic surges in baroreflex failure. PMID- 8413456 TI - The effect of angiotensin-converting-enzyme inhibition on diabetic nephropathy. The Collaborative Study Group. AB - BACKGROUND: Renal function declines progressively in patients who have diabetic nephropathy, and the decline may be slowed by antihypertensive drugs. The purpose of this study was to determine whether captopril has kidney-protecting properties independent of its effect on blood pressure in diabetic nephropathy. METHODS: We performed a randomized, controlled trial comparing captopril with placebo in patients with insulin-dependent diabetes mellitus in whom urinary protein excretion was > or = 500 mg per day and the serum creatinine concentration was < or = 2.5 mg per deciliter (221 mumol per liter). Blood-pressure goals were defined to achieve control during a median follow-up of three years. The primary end point was a doubling of the base-line serum creatinine concentration. RESULTS: Two hundred seven patients received captopril, and 202 placebo. Serum creatinine concentrations doubled in 25 patients in the captopril group, as compared with 43 patients in the placebo group (P = 0.007). The associated reductions in risk of a doubling of the serum creatinine concentration were 48 percent in the captopril group as a whole, 76 percent in the subgroup with a baseline serum creatinine concentration of 2.0 mg per deciliter (177 mumol per liter), 55 percent in the subgroup with a concentration of 1.5 mg per deciliter (133 mumol per liter), and 17 percent in the subgroup with a concentration of 1.0 mg per deciliter (88.4 mumol per liter). The mean (+/- SD) rate of decline in creatinine clearance was 11 +/- 21 percent per year in the captopril group and 17 +/- 20 percent per year in the placebo group (P = 0.03). Among the patients whose base-line serum creatinine concentration was > or = 1.5 mg per deciliter, creatinine clearance declined at a rate of 23 +/- 25 percent per year in the captopril group and at a rate of 37 +/- 25 percent per year in the placebo group (P = 0.01). Captopril treatment was associated with a 50 percent reduction in the risk of the combined end points of death, dialysis, and transplantation that was independent of the small disparity in blood pressure between the groups. CONCLUSIONS: Captopril protects against deterioration in renal function in insulin-dependent diabetic nephropathy and is significantly more effective than blood-pressure control alone. PMID- 8413457 TI - Fetal thrombocytopenia and its relation to maternal thrombocytopenia. AB - BACKGROUND: Neonates with severe thrombocytopenia can have bleeding leading to death or lifelong residual defects. The predictors, frequency, and consequences of fetal thrombocytopenia are not known, nor is it known if there are maternal clinical features that could predict fetal thrombocytopenia. METHODS: We conducted a seven-year cross-sectional study in which platelet counts were determined in newborns' umbilical-cord blood and blood obtained from their mothers at consecutive deliveries in one obstetrical unit. The relations of the umbilical-cord platelet count to maternal risk factors were determined. RESULTS: Platelet counts were determined in blood samples from 15,471 mothers and 15,932 newborn infants. The cord-blood platelet count was less than 50,000 per cubic millimeter in 19 infants (0.12 percent; 95 percent confidence interval, 0.07 to 0.19 percent), whereas the platelet count was less than 150,000 per cubic millimeter in 6.6 percent of the mothers (95 percent confidence interval, 6.2 to 7.0 percent). One infant among those born to 756 mothers with incidental thrombocytopenia, 5 infants among those born to 1414 mothers with hypertension, and 4 infants among those born to 46 mothers with idiopathic thrombocytopenic purpura had cord-blood platelet counts between 20,000 and 50,000 per cubic millimeter. Only 6 infants (0.04 percent; 95 percent confidence interval, 0.01 to 0.08 percent) had cord-blood platelet counts of less than 20,000 per cubic millimeter; all their mothers were among the 18 whose 19 fetuses were at risk for neonatal alloimmune thrombocytopenia. Two of these infants had in utero intracranial hemorrhage. In addition, 3 infants born to these 18 women had cord blood platelet counts between 20,000 and 50,000 per cubic millimeter; there was 1 stillbirth due to intracranial hemorrhage. CONCLUSIONS: Moderate-to-severe fetal thrombocytopenia is a rare event. The only severely affected neonates with morbidity or mortality due to this condition are those born to mothers with antiplatelet alloantibodies. PMID- 8413458 TI - Images in clinical medicine. Pseudothrombocytopenia. PMID- 8413459 TI - Pathogenesis of Graves' ophthalmopathy. PMID- 8413460 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 45-1993. A 23-year-old asthmatic man with pulmonary infiltrates and hilar lymphadenopathy. PMID- 8413461 TI - Infants with fever. PMID- 8413462 TI - Baroreflex failure--a diagnostic challenge. PMID- 8413463 TI - Slowing the progression of diabetic nephropathy. PMID- 8413465 TI - Fetal-tissue transplantation for Parkinson's disease. PMID- 8413464 TI - Fetal-tissue transplantation for Parkinson's disease. PMID- 8413466 TI - Fetal-tissue transplantation for Parkinson's disease. PMID- 8413467 TI - Pancreatitis and the risk of pancreatic cancer. PMID- 8413468 TI - Tretinoin for hyperpigmentation in black patients. PMID- 8413469 TI - Visceral leishmaniasis in Desert Storm veterans. PMID- 8413470 TI - Cyanobacterium-like cyclospora species. PMID- 8413471 TI - Cyanobacterium-like cyclospora species. PMID- 8413472 TI - Clinical light damage by indirect ophthalmoscopy. PMID- 8413473 TI - An update on the vitamin D content of fortified milk from the United States and Canada. PMID- 8413474 TI - Increase in suicide by asphyxiation in New York City after the publication of Final Exit. PMID- 8413475 TI - Prophylactic administration of respiratory syncytial virus immune globulin to high-risk infants and young children. The Respiratory Syncytial Virus Immune Globulin Study Group. AB - BACKGROUND: Infants with cardiac disease or prematurity are at risk for severe illness caused by respiratory syncytial virus. Immune globulin with a high titer of antibodies against respiratory syncytial virus may offer infants and young children at risk protection from this serious, common respiratory illness. METHODS: We studied 249 infants and young children (mean age, eight months) who had bronchopulmonary dysplasia due to prematurity (n = 102), congenital heart disease (n = 87), or prematurity alone (n = 60). Respiratory syncytial virus immune globulin was given monthly to some of these children in either a high dose (750 mg per kilogram of body weight; n = 81) or low dose (150 mg per kilogram; n = 79); 89 controls received no immune globulin. Group assignments were random. Assessments of respiratory illness and management were conducted without knowledge of the children's group assignments. RESULTS: There were 64 episodes of respiratory syncytial virus infection: 19 in the high-dose group, 16 in the low dose group, and 29 in the control group. In the high-dose group there were fewer lower respiratory tract infections (7, vs. 20 in the control group; P = 0.01), fewer hospitalizations (6, vs. 18 in the control group; P = 0.02), fewer hospital days (43, vs. 128 in the control group; P = 0.02), fewer days in the intensive care unit (P = 0.05), and less use of ribavirin (P = 0.05). In the low-dose group there was a significant reduction only in the number of days in the intensive care unit (P = 0.03). Adverse events during the 580 infusions were generally mild and included fluid overload (in five children), oxygen desaturation (eight), and fever (six). Six children died: three in the high-dose group, three in the low dose group, and none in the control group (P = 0.15), but no death was attributed to the use of immune globulin or to illness caused by respiratory syncytial virus. CONCLUSIONS: Administration of high doses of respiratory syncytial virus immune globulin is a safe and effective means of preventing lower respiratory tract infection in infants and young children at high risk for this disease. PMID- 8413476 TI - Elevated serum inhibin concentrations in postmenopausal women with ovarian tumors. AB - BACKGROUND: Inhibin is an ovarian hormone that inhibits the secretion of follicle stimulating hormone (FSH) by the anterior pituitary gland. Women with granulosa cell tumors of the ovary have elevated serum inhibin concentrations, but whether the concentrations are increased in women with other ovarian tumors is unknown. METHODS: We measured serum inhibin and FSH concentrations before surgery in 212 postmenopausal women with suspected ovarian cancer and after surgery in 210 of them. RESULTS: Eighteen of the 22 women (82 percent) with mucinous carcinomas (mucinous cystadenocarcinomas and mucinous borderline cystic tumors) of the ovary had elevated serum inhibin concentrations, whereas only 9 of the 53 women (17 percent) with serous carcinomas (serous cystadenocarcinomas and serous borderline cystic tumors) had elevated levels. Serum inhibin concentrations were also elevated in 2 of 12 women (17 percent) with clear-cell carcinomas, 4 of 26 women (15 percent) with undifferentiated carcinomas, 3 of 3 women (100 percent) with granulosa-cell tumors, and 5 of 27 women (19 percent) with other ovarian cancers. The serum concentrations of inhibin were increased in 2 of 28 women (7 percent) with nonovarian pelvic cancers and 11 of 41 women (27 percent) with benign ovarian diseases. All women but one with initially elevated serum inhibin concentrations had low values one week after surgery. Serum inhibin concentrations correlated negatively with serum FSH concentrations (P = 0.05) in women with granulosa-cell tumors but not in women with other tumors, suggesting that the inhibin secreted by tumors in the latter group has decreased biologic activity. CONCLUSIONS: Serum inhibin concentrations are elevated in most postmenopausal women with mucinous carcinomas of the ovary and in some women with other types of epithelial ovarian tumors. The concentrations fall after tumor removal. PMID- 8413477 TI - A tobacco-specific lung carcinogen in the urine of men exposed to cigarette smoke. AB - BACKGROUND: Environmental tobacco smoke has been classified by the Environmental Protection Agency as a carcinogen causally associated with lung cancer in adults, but there have been no reports of lung carcinogens or their metabolites in the body fluids or tissues of nonsmokers exposed to environmental tobacco smoke. METHODS: Five male nonsmokers were exposed to sidestream cigarette smoke generated by machine smoking of reference cigarettes for 180 minutes on each of two days, six months apart. Sidestream smoke is the smoke that originates from the smoldering end of a cigarette between puffs. Twenty-four-hour urine samples were collected before and after exposure. The urine samples were analyzed for 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its glucuronide, which are metabolites of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a powerful lung carcinogen in rodents. NNAL is also a lung carcinogen in rodents. RESULTS: The urinary excretion of the metabolites increased after exposure to sidestream smoke in all the men. The mean (+/- SD) amount of NNAL and NNAL glucuronide was significantly higher after exposure than at base line (33.9 +/- 20.0 vs. 8.4 +/- 11.2 ng per 24 hours [127 +/- 74 vs. 31 +/- 41 pmol per day], P < 0.001) and was correlated with urinary cotinine excretion (r = 0.89, P < 0.001). The nicotine concentrations in the air to which the men were exposed were comparable to those in a heavily smoke-polluted bar. CONCLUSIONS: Nonsmokers exposed to sidestream cigarette smoke take up and metabolize a lung carcinogen, which provides experimental support for the proposal that environmental tobacco smoke can cause lung cancer. PMID- 8413478 TI - Images in clinical medicine. Neurofibromatosis type 1. PMID- 8413479 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 46-1993. A 75-year-old man with right-sided rigidity, dysarthria, and abnormal gait. PMID- 8413480 TI - The beginning of health care reform: the Clinton plan. PMID- 8413481 TI - The ductus arteriosus and its closure. PMID- 8413482 TI - Respiratory syncytial virus--successful immunoprophylaxis at last. PMID- 8413483 TI - Radiation therapy for in situ or localized breast cancer. PMID- 8413484 TI - Radiation therapy for in situ or localized breast cancer. PMID- 8413485 TI - Radiation therapy for in situ or localized breast cancer. PMID- 8413486 TI - Vitamins and breast cancer. PMID- 8413487 TI - The hazards of active and passive smoking. PMID- 8413488 TI - The hazards of active and passive smoking. PMID- 8413489 TI - More on the nicotine content of vegetables. PMID- 8413490 TI - Cutaneous disease and drug reactions in HIV infection. PMID- 8413491 TI - Chronic urticaria due to surgical clips. PMID- 8413492 TI - Prevention and treatment of traveler's diarrhea. PMID- 8413493 TI - Prevention and treatment of traveler's diarrhea. PMID- 8413494 TI - Prevention and treatment of traveler's diarrhea. PMID- 8413495 TI - Health care reform. The labyrinth of Congress. PMID- 8413496 TI - Experimental pathogenicity of Sporothrix schenckii preserved in water (Castellani). AB - Five strains of Sporothrix schenckii preserved in sterile distilled water were found morphologically stable, viable and pure after a period ranging from 16 to 13 years, in the mycology section culture collection from the Instituto de Medicina Tropical, Universidad Central de Venezuela. Each strain was inoculated into the testes of 3 hamsters. All the animals developed disseminated sporotrichosis. PMID- 8413497 TI - Effects of antimycotics on the biosynthesis of cellular macromolecules in Aspergillus niger protoplasts. AB - The effects of nine antimycotics on the biosynthesis of cellular macromolecules were analyzed using the regenerating system of protoplasts of Aspergillus niger. The incorporation of several specific radioactive precursors into major cellular components were measured in the presence or in the absence of respective agents. Miconazole, ketoconazole, and tolnaftate inhibited the lipid synthesis. 5 Fluorocytosine strongly inhibited the DNA and protein syntheses. Griseofulvin, however, specifically inhibited the synthesis of cell wall polysaccharides, i.e. chitin and glucan. Other agents showed non-specific inhibition effects. The significance of morphological change of hypha as an indicator of antimycotic action and its feasibility as a screening tool for novel antimycotic compounds are discussed. PMID- 8413498 TI - Epidemiology of the dermatophytoses in the Florence area of Italy: 1985-1990. Trichophyton mentagrophytes, Epidermophyton floccosum and Microsporum gypseum infections. AB - Between 1985 and 1990 we observed 2085 cases of dermatophytoses in the Florence area of Italy. Trichophyton rubrum and Microsporum canis were the major etiological agents. 126 cases of dermatophytoses observed during this period were caused by Trichophyton mentagrophytes. Most of the patients came from rural areas or they kept pets. Epidermophyton floccosum ranked fourth in frequency (95 cases). The patients were adults and mainly males, most of whom engaged in sports or lived in group environments. Microsporum gypseum was isolated in 31 cases. Infections occurred directly from contact with soil or domestic animals. We report the principal clinical data concerning these last three dermatophyte infections. PMID- 8413499 TI - Keratinophilic fungi in the antarctic environment. AB - Results of a study on the diffusion of keratinophilic fungi in an Antarctic environment are given. Nine soil samples collected from as many sites along the coast of Ross Sea, and six dust samples inside the Italian scientific base were examined by direct inoculation and hair baiting methods for soil samples and plate dilution method for dust samples. As regards the variety of species isolated and the counting of the numbers of colonies, plate dilution method proved to be very effective. Aspergillus spp., Cladosporium sp., Dematiaceae, Fusarium sp., Geomyces pannorum v. pannorum, G. pannorum v. vinaceus, Mycelia sterilia, Penicillium spp. were isolated from soil. From the dust the following moulds were isolated: Aphanoascus fulvescens, Aspergillus sp., Beauveria sp., Chrysosporium carmichaelii, Dematiaceae, Geomyces pannorum v. pannorum, G. pannorum v. vinaceus, Malbranchea gypsea, Mycelia sterilia, Nectria inventa, Penicillium spp., Scopulariopsis brevicaulis, Scopulariopsis sp. and Trichophyton mentagrophytes. The presence of four colonies of Trichophyton mentagrophytes is emphasized and correlated with the anthropization process. PMID- 8413500 TI - Mycotoxigenic potential of Aspergillus flavus strains isolated from groundnuts growing in Israel. AB - Two hundred strains of the Aspergillus flavus group isolated from groundnuts (peanuts) growing in Israel were examined for their ability to produce mycotoxins in potato dextrose (PD) broth. Almost 77% of the isolates produced aflatoxin; aflatoxins B1 and B2 were formed by most of the isolates. Simultaneous production of aflatoxins of groups B and G was detected in only 0.5% of the isolates. Microscopic examination revealed that 98% of the isolates were A. flavus and only 2% A. parasiticus. Cyclopiazonic acid (CPA) was detected in 22.5% of the isolates, including 3.5% that produced only CPA. Sterigmatocystin was detected in only 2% of the isolates and only one isolate produced aflatoxin simultaneously with CPA and sterigmatocystin. The dry weight (DW) of mycelium, 7 days after inoculating the medium, was between 71-110 mg/30 ml medium in more than 70% of the isolates. A general decrease in the pH was observed and 75% of the isolates reduced the pH to 4.5 or below. After 14 days, a small increase in DW and an increase in the pH toward neutrality was observed. On PD agar, 30% of the isolates produced sclerotia, including 5% that produced them profusely. No correlation between mycelial growth, changes in pH of the medium, sclerotium formation, and aflatoxin accumulation could be observed. The mycotoxigenic potential of the A. flavus strains isolated from groundnuts seems to be relatively high and may present a potential threat to human and animal health. PMID- 8413501 TI - Occurrence and toxicity of Fusarium subglutinans from Peruvian maize. AB - Twenty-five samples of maize kernels collected at harvest time from geographically different corn fields in Peru, were examined for the occurrence of toxigenic Fusarium species. The most frequently recovered species were F. subglutinans (48%), F. moniliforme (46%), and F. equiseti (5%). Other Fusarium species isolated (up to 1%) included F. graminearum, F. acuminatum, F. solani, F. oxysporum, and F. culmorum. Assays of Fusarium culture extracts using Artemia salina larvae, showed F. subglutinans as one of the most toxigenic species, and its toxicity was mostly correlated to the capability to produce beauvericin (BEA). All eight tested isolates of F. subglutinans grown on autoclaved corn kernels produced BEA (from 50 to 250 mg/kg) as well as moniliformin (M) (from 70 to 270 mg/Kg). This is the first report on BEA and M production by maize isolates of F. subglutinans from South America. PMID- 8413502 TI - Drug abuse among minority youth: methodological issues and recent research advances. July 17-18, 1991, Bethesda, Maryland. Proceedings. PMID- 8413503 TI - Network theory: a model for understanding drug abuse among African-American and Hispanic youth. PMID- 8413504 TI - Examining conceptual models for understanding drug use behavior among American Indian youth. PMID- 8413505 TI - Acculturation strain theory: its application in explaining drug use behavior among Cuban and other Hispanic youth. PMID- 8413506 TI - Validity of self-reports in student-based studies on minority populations: issues and concerns. PMID- 8413507 TI - Interviewing minority youth about drug use: telephone vs. in-person surveys. PMID- 8413508 TI - Hispanic dropouts and drug use: a review of the literature and methodological considerations. PMID- 8413509 TI - Getting into the gang: methodological issues in studying ethnic gangs. PMID- 8413510 TI - Identifying, gaining access to, and collecting data on African-American drug addicts. PMID- 8413511 TI - Surveying and tracking urban elementary school children's use of abusable substances. PMID- 8413512 TI - School and community politics: issues, concerns, and implications when conducting research in African-American communities. PMID- 8413513 TI - Substance use disorders among young minority refugees: common themes in a clinical sample. PMID- 8413514 TI - Orthogonal cultural identification: theoretical links between cultural identification and substance use. PMID- 8413515 TI - Current gaps and new directions for studying drug use and abuse behavior in minority youth. PMID- 8413516 TI - Acculturation: the broader view. Theoretical framework of the acculturation scales. PMID- 8413517 TI - Interactional theory: its utility in explaining drug use behavior among African American and Puerto Rican youth. PMID- 8413518 TI - Integrating mainstream and subcultural explanations of drug use among Puerto Rican youth. PMID- 8413519 TI - The impact of prescription drug control systems on medical practice and patient care: a summary of the NIDA technical review. PMID- 8413520 TI - Impact of prescription drug diversion control systems on medical practice and patient care. Technical review, May 30-June 1, 1991. Proceedings. PMID- 8413521 TI - The nonmedical use of prescription drugs in the United States. PMID- 8413522 TI - Drug diversion control systems, medical practice, and patient care. PMID- 8413523 TI - Existing methods to identify retail drug diversion. PMID- 8413524 TI - PADS approach to preventing prescription drug diversion. PMID- 8413525 TI - Diversion investigation units--methods, utilities, and limitations. PMID- 8413526 TI - OSTAR-Oklahoma Schedule II abuse reduction: an electronic point of sale diversion control system. AB - In its short history, OSTAR has proven to be a fast, accurate tracking system for Schedule II prescriptions within Oklahoma. Information is available immediately in many cases and, having been entered into the system by the pharmacist, is very accurate. Approximately 50 percent of the information is being transmitted through electronic means at this time, and that percentage is increasing daily due to software companies making modifications to their pharmacy programs. System deficiencies have proven to be minimal, even less than anticipated. Efforts are still underway to ensure that data is received in a uniform manner; i.e., some data systems include zeroes at the end of the patient number, which causes investigators to have to query in more than one form. These pharmacies and companies are currently being notified and are making the necessary changes. The technical problems usually encountered with a new system have been limited, and data has been useful from the first month of the program. Feedback from impaired physicians has indicated they would not have been so free to divert Schedule II substances through prescriptions if this system had been in place. OBN has active cases involving a large number of physician scammers that are a direct result of OSTAR and it has enhanced cases on at least one physician and provided valuable information for search warrants. Agents are able to make better cases for prosecutors while completing investigations in much shorter periods of time. Local law enforcement personnel who have accessed OSTAR have become convinced that it is a most effective tool, and use of the system has complemented their investigations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413527 TI - The Illinois experience in achieving the medical/regulatory balance required to control prescription drug diversion. PMID- 8413528 TI - New York State's Triplicate Prescription Program. PMID- 8413529 TI - Guiding principles of international and federal laws pertaining to medical use and diversion of controlled substances. PMID- 8413530 TI - Triplicate prescriptions in Washington State. AB - In summary, I believe that this program, although limited in scope, has been of value to our State in controlling the inappropriate prescribing of controlled substances. We have identified several problems that have been or are being addressed in order to make the program even more effective. The program targets only those prescribers who have been identified as having drug use or prescribing problems rather than subjecting all practitioners to a program for the purpose of finding a relatively small number of violators. Thus, the intimidation factor alleged to be present in full triplicate programs is absent in the limited program. It is flexible since the drugs included in the monitoring can range from a single controlled substance schedule, all schedules, or even to all prescriptions. The program includes both prescribed drugs and those administered in the office or dispensed by the practitioner. The responsibility for compliance is placed on the prescribers, not on the pharmacists. The way the law is written, the program could be expanded to include a sample of all prescribers, providing information that could be used to determine prescribing norms. Overprescribing appears to be in the eyes of the beholder. A pharmacist may view certain prescribing habits differently than a disciplinary board. The availability of norms would assist boards in making more scientific determinations regarding alleged overprescribing practices. PMID- 8413531 TI - The Medicaid Prescription Drug Initiative. PMID- 8413532 TI - Summary and conclusions of a review of prescription drug diversion control methods. PMID- 8413533 TI - An American Medical Association perspective on preventing prescription drug diversion. PMID- 8413534 TI - Prescription drug regulation: implications for nursing and health care delivery- response of the American Nurses' Association. PMID- 8413535 TI - Perspective of the American Pharmaceutical Association. PMID- 8413536 TI - Perspective of the Empire State Medical Association of the National Medical Association. PMID- 8413537 TI - Prescribing practices and drug abuse--perspective of the American Society of Addiction Medicine. PMID- 8413538 TI - Response of the American Academy of Child and Adolescent Psychiatry. PMID- 8413539 TI - State cancer pain initiatives. PMID- 8413540 TI - A Public Citizen Health Research Group perspective on federal triplicate prescription requirements for controlled substance prescription drugs. PMID- 8413541 TI - A research agenda for prescription drug diversion control. PMID- 8413542 TI - The impact of prescription drug control systems on medical practice and patient care--a summary of research recommendations. PMID- 8413543 TI - Therapeutic use of opioids: prescribing and control issues. PMID- 8413544 TI - Issues and controversies regarding benzodiazepine use. PMID- 8413545 TI - News on accident and emergency nursing. PMID- 8413546 TI - Minimising the hazards. PMID- 8413547 TI - AIDS. PMID- 8413548 TI - Pitfalls in the management of acute post-operative pain. PMID- 8413549 TI - Taking stock of midwifery. PMID- 8413550 TI - Understanding aggression in a child. PMID- 8413551 TI - Which NSAID? PMID- 8413552 TI - [Obtaining an adequate Pap cell test]. AB - Routine use of the Papanicolaou (Pap) smear has led to a decrease in mortality from cervical carcinoma. The Papanicolaou test examines exfoliated cells to detect preinvasive lesions (eg, dysplasia, carcinoma-in-situ) as well as invasive lesions. Cervical screening is recommended for women who are sexually active regardless of age. For the woman who is not sexually active, cervical screening should begin at 18 years of age. An essential part of a good Pap smear technique is that the specimen should consist of cells from the squamocolumnar junction. It is important that the specimen should be fixed quickly. PMID- 8413553 TI - My experience as a DEPAM facilitator (Decentralised Education Programme for Advanced Midwifery). PMID- 8413554 TI - Sexual harassment in nursing. PMID- 8413555 TI - Didanosine therapy in patients with HIV infection. PMID- 8413556 TI - Modern options in pain management. PMID- 8413557 TI - [X-rays taken by nurses]. AB - Medical technology makes use of ionising radiation in many ways to diagnose and treat disease. One of these is X-rays. In rural areas there are hospitals where the services of a radiographer are not available and nurses take over this task. No formal training is available for these nurses in order to equip them with the necessary knowledge and skills to take X-rays. This article looks into the nature of this phenomenon. PMID- 8413558 TI - South Africa and AIDS: seven years wasted. PMID- 8413559 TI - AIDS. The bumblers play on--a decade of debacle. PMID- 8413560 TI - Report from AIDS Conference 1992. PMID- 8413562 TI - Mentoring. PMID- 8413561 TI - [AIDS: ethical implications]. AB - Immunodeficiency syndrome or AIDS is a reality in South Africa and poses an urgent challenge to nurses. The caring for patients with AIDS and the prevention of AIDS has ethical implications for nurses. Professional secrecy, social stigma, the attitude of nurses and judicial aspects are analysed. Nurses from the frontline of health care givers and must take an active part to prevent AIDS from spreading. PMID- 8413563 TI - Nurses in tuberculosis control. PMID- 8413564 TI - The integration of research and practice. PMID- 8413565 TI - [Legal protection of the mentally ill]. PMID- 8413566 TI - Budesonide--effective treatment in newly detected asthmatics. PMID- 8413567 TI - Shaping a new nursing organization. PMID- 8413569 TI - Coping with death. PMID- 8413568 TI - The profession. New ways to bottle old wine. PMID- 8413570 TI - Cancer and the rural black family. PMID- 8413571 TI - [Why does the patient with cancer have malnutrition?]. AB - The effective nursing management of the nutritional needs of patients with carcinoma demands not only knowledge of the causes of malnutrition, but also that the nurse will consider these causes in her planning of nursing interventions. PMID- 8413572 TI - Tuberculosis control. Part 2: Process of diagnosis. PMID- 8413574 TI - What's in a name? PMID- 8413573 TI - Understanding laboratory and diagnostic tests. PMID- 8413575 TI - Activists now urge caution on approval of new AIDS drug. PMID- 8413576 TI - US plans to abandon Delaney Clause. PMID- 8413577 TI - Academies urge population control. PMID- 8413578 TI - Brain institute falls victim to war in ex-Yugoslavia. PMID- 8413579 TI - Scientists protest at 'clean' appointments. PMID- 8413580 TI - Whose research is it? PMID- 8413581 TI - Cell signalling. A tale of two messengers. PMID- 8413582 TI - Dopamine receptors. The D4 and schizophrenia. PMID- 8413583 TI - HIV. Cyclophilins unfold the Gag? PMID- 8413584 TI - New roles for G-protein beta gamma-dimers in transmembrane signalling. AB - When a membrane-bound receptor acts on a G protein, the GTP-binding or G alpha subunit dissociates from the G beta gamma dimer. Until recently, the G alpha subunit alone was thought to act on the enzymes and ion channels controlled by these proteins. Newer evidence indicates that the G beta gamma dimer also plays a major part in signal transmission, enhancing the complexity of the possible interactions between the G proteins and their targets. PMID- 8413585 TI - Association between proto-oncoprotein Rel and TATA-binding protein mediates transcriptional activation by NF-kappa B. AB - The c-Rel protein is able to associate in vitro and in vivo with the TATA-binding protein (TBP) of the TFIID complex. Coexpression of TBP with c-Rel augments transactivation from the kappa B site in Drosophila Schneider cells. DNA-binding mutants of TBP not only fail to cooperate, but they repress transactivation by c Rel. There may be a direct communication between kappa B enhancer binding proteins and basal transcription factors which leads to enhanced transcription. PMID- 8413586 TI - Telling tails explain the discrepancy in sexual partner reports. AB - An anomaly often noted in surveys of sexual behaviour is that the number of female sexual partners reported by men exceeds the number of male partners reported by women. This discrepancy is sometimes interpreted as evidence that surveys produce unreliable data due to sex-linked response and sampling bias. We report here that among the 90% of respondents reporting fewer than 20 lifetime partners, however, the ratio of male to female reports drops from 3.2:1 to 1.2:1. The anomaly thus appears to be driven by the upper tail of the contact distribution, an example of the general principle of outlier influence in data analysis. The implication is that sexual behaviour surveys provide reliable data in the main, and that simple improvements can increase precision in the upper tail to make these data more useful for modelling the spread of AIDS and other sexually transmitted diseases. PMID- 8413587 TI - Dopamine D4 receptors elevated in schizophrenia. AB - Although the biological basis of schizophrenia is not known, possible causes include genetic defects, viruses, amines, brain structure and metabolism, neuroreceptors, and G proteins. The hypothesis of dopamine overactivity in schizophrenia is based on the fact that neuroleptics block dopamine D2 receptors in direct relation to their clinical antipsychotic potencies. Moreover, dopamine D2 or D2-like receptors are elevated in postmortem schizophrenia brain tissue. This elevation, however, is only found in vivo using [11C]methylspiperone but not [11C]raclopride. The dopamine D4 receptor gene has not yet been excluded in schizophrenia because the 21 gene variants of D4 have not yet been tested. Because the link between D1 and D2 receptors is reduced in schizophrenia tissue, we tested whether one component of this link was sensitive to guanine nucleotide. We report here that the binding of [3H]raclopride to D2 receptors in schizophrenia was not sensitive to guanine nucleotide. This finding permitted analysis of data on the binding of [3H]emonapride to the D2, D3 and D4 receptors. We conclude that the combined density of D2 and D3 receptors (labelled by [3H]raclopride) is increased by only 10% in schizophrenia brain, as found by Farde et al., but that it is the density of dopamine D4 receptors which is sixfold elevated in schizophrenia. These findings resolve the apparent discrepancy, mentioned above, wherein the density of [11C]methylspiperone labelled sites (D2, D3 and D4), but not that of [11C]raclopride-labelled sites (D2 and D3), was found elevated in the schizophrenia striatum. PMID- 8413588 TI - Generating loss-of-function phenotypes of the fushi tarazu gene with a targeted ribozyme in Drosophila. AB - The ability to isolate gene sequences and analyse their expression patterns has generated demand for mutations created to assess their biological functions. In Drosophila melanogaster this can be achieved by traditional mutagenesis, but this is time-consuming, labour-intensive and not always successful. Moreover, the functions of genes that are expressed several times during development are often obscured in the later stages because of disruptions caused by the absence of early gene function. Here we propose a new strategy to create conditional knock out mutations using a targeted heat-inducible ribozyme. Ribozymes are catalytic RNA molecules that specifically cleave RNAs and are potentially useful for studying gene function during animal development because the expression of critical regulatory genes is usually low and their function is often dosage dependent. The ribozyme can be delivered to a specific region or at a particular developmental stage using a region-specific or inducible promoter. The Drosophila fushi tarazu (ftz) gene is a good candidate for testing this approach. We generated transgenic flies carrying a ribozyme against the ftz gene. The two developmental phases of ftz function can be distinguished by timed induction of the ribozyme. Activation of the ribozyme in the blastoderm disrupts the ftz seven stripe pattern and produces ftz-like pair-rule defects in larvae. The involvement of ftz in neurogenesis was verified by activation of the ribozyme during the early phase of formation of the central nervous system. PMID- 8413589 TI - Skeletal muscle myosin light chains are essential for physiological speeds of shortening. AB - In muscle each myosin head contains a regulatory light chain (LC2) that is wrapped around the head/rod junction, and an alkali light chain that is distal to LC2 (ref. 1). The role of these light chains in vertebrate skeletal muscle myosin has remained obscure. Here we prepare heavy chains that are free of both light chains in order to determine by a motility assay whether the light chains are necessary for movement. We find that removal of light chains from myosin reduces the velocity of actin filaments from 8.8 microns s-1 to 0.8 microns s-1 without significantly decreasing the ATPase activity. Reconstitution of myosin with LC2 or alkali light chain increases filament velocity to intermediate rates, and readdition of both classes of light chains fully restores the original sliding velocity. We conclude that even though the light chains are not essential for enzymatic activity, light-chain/heavy-chain interactions play an important part in the conversion of chemical energy into movement. PMID- 8413590 TI - Valosin-containing protein, VCP, is a ubiquitous clathrin-binding protein. AB - Clathrin is the structural protein of coated membranes involved in receptor mediated endocytosis and aspects of Golgi sorting in eukaryotic cells. We have now detected a stoichiometric complex of clathrin with a novel protein of M(r) approximately 100,000 (100K) in lysates of different mammalian cells. Formation of the complex, which also includes the 70K heat-shock protein Hsc70, occurs within 15 min of synthesis. The 100K protein has been identified as valosin containing protein (VCP; ref. 1), an early substrate for tyrosine phosphorylation on T-cell receptor activation. Further, VCP is the mammalian homologue of yeast Cdc48p (ref. 3) and is a member of a larger gene family that includes putative ATP-binding proteins involved in vesicle transport and fusion, 26S proteasome function, regulation of the expression of human immunodeficiency virus, and assembly of peroxisomes. The association with clathrin and the morphological and catalytic similarity to the chaperonin proteins indicate that VCP may modulate protein-protein interactions in membrane transport processes. PMID- 8413591 TI - Crystal structure of the DsbA protein required for disulphide bond formation in vivo. AB - Proteins that contain disulphide bonds are often slow to fold in vitro because the oxidation and correct pairing of the cysteine residues is rate limiting. The folding of such proteins is greatly accelerated in Escherichia coli by DsbA, but the mechanism of this rate enhancement is not well understood. Here we report the crystal structure of oxidized DsbA and show that it resembles closely the ubiquitous redox protein thioredoxin, despite very low sequence similarity. An important difference, however, is the presence of another domain which forms a cap over the thioredoxin-like active site of DsbA. The redox-active disulphide bond, which is responsible for the oxidation of substrates, is thus at a domain interface and is surrounded by grooves and exposed hydrophobic side chains. These features suggest that DsbA might act by binding to partially folded polypeptide chains before oxidation of cysteine residues. PMID- 8413592 TI - A new component of the transcription factor DRTF1/E2F. PMID- 8413593 TI - Parallel genome analysis by two-dimensional DNA typing. AB - By two-dimensional (2-D) DNA typing a restriction enzyme digest of genomic DNA can be resolved on the basis of both size and base-pair sequence and subsequently analysed by repeat probe hybridization to reveal sequence variants at multiple genomic sites in parallel. The system has been partly automated and allows for large-scale comparative analysis of complex genomes in a cost-effective manner. PMID- 8413594 TI - MRC research centre under threat from London health service plans. PMID- 8413595 TI - Rhone-Poulenc lifts Institut Merieux stake. PMID- 8413596 TI - Army AIDS vaccine trial faces new block. PMID- 8413597 TI - First anthropoid station at risk. PMID- 8413598 TI - Transcription. Opening the gateway. PMID- 8413599 TI - Peptide engineering. Catalytic molten globules. PMID- 8413600 TI - Nucleic acid recognition. Marriage of convenience. PMID- 8413601 TI - Palaeoanthropology. Renaissance of Europe's ape. PMID- 8413602 TI - Fatty acylated toxin structure. PMID- 8413603 TI - Perception of heading. PMID- 8413604 TI - Crystal structure of a yeast TBP/TATA-box complex. AB - The 2.5 A crystal structure of a TATA-box complex with yeast TBP shows that the eight base pairs of the TATA box bind to the concave surface of TBP by bending towards the major groove with unprecedented severity. This produces a wide open, underwound, shallow minor groove which forms a primarily hydrophobic interface with the entire under-surface of the TBP saddle. The severe bend and a positive writhe radically alter the trajectory of the flanking B-form DNA. PMID- 8413605 TI - Co-crystal structure of TBP recognizing the minor groove of a TATA element. AB - The three-dimensional structure of a TATA-box binding polypeptide complexed with the TATA element of the adenovirus major late promoter has been determined by X ray crystallography at 2.25 A resolution. Binding of the saddle-shaped protein induces a conformational change in the DNA, inducing sharp kinks at either end of the sequence TATAAAAG. Between the kinks, the right-handed double helix is smoothly curved and partially unwound, presenting a widened minor groove to TBP's concave, antiparallel beta-sheet. Side-chain/base interactions are restricted to the minor groove, and include hydrogen bonds, van der Waals contacts and phenylalanine-base stacking interactions. PMID- 8413606 TI - Synthesis, structure and activity of artificial, rationally designed catalytic polypeptides. AB - Biological macromolecules with catalytic activity can be created artificially using two approaches. The first exploits a system that selects a few catalytically active biomolecules from a large pool of randomly generated (and largely inactive) molecules. Catalytic antibodies and many catalytic RNA molecules are obtained in this way. The second involves rational design of a biomolecule that folds in solution to present to the substrate an array of catalytic functional groups. Here we report the synthesis of rationally designed polypeptides that catalyse the decarboxylation of oxaloacetate via an imine intermediate. We determine the secondary structures of the polypeptides by two dimensional NMR spectroscopy. We are able to trap and identify intermediates in the catalytic cycle, and to explore the kinetics in detail. The formation of the imine by our artificial oxaloacetate decarboxylases is three to four orders of magnitude faster than can be achieved with simple amine catalysts: this performance rivals that of typical catalytic antibodies. PMID- 8413607 TI - Recent discoveries of Dryopithecus shed new light on evolution of great apes. AB - The origin and early evolution of the great ape/human clade (Hominidae) is currently a subject of debate. The controversy is fuelled by the fragmentary nature of the fossils which renders it difficult to determine clearly derived features that permit the recognition of fossil members of this clade. We report here the recent discovery of a facial skeleton and a temporal fragment with the petrosal bone of Dryopithecus laietanus, which provides a way out of an impasse. The lack of the fossa subarcuata is a great ape and human clade synapomorphy, and proves unequivocally that Dryopithecus belongs to this clade. The zygomatic possesses derived characters which reveal that Dryopithecus is related to the Ponginae and not to the African apes/humans, as recently suggested. The remaining morphological features are plesiomorphic and thus provide a good model of a common ancestor of all Hominidae. PMID- 8413608 TI - A role for central vasopressin in pair bonding in monogamous prairie voles. AB - Monogamous social organization is characterized by selective affiliation with a partner, high levels of paternal behaviour and, in many species, intense aggression towards strangers for defence of territory, nest and mate. Although much has been written about the evolutionary causes of monogamy, little is known about the proximate mechanisms for pair bonding in monogamous mammals. The prairie vole, Microtus ochrogaster, is a monogamous, biparental rodent which exhibits long-term pair bonds characterized by selective affiliation (partner preference) and aggression. Here we describe the rapid development of both selective aggression and partner preferences following mating in the male of this species. We hypothesized that either arginine-vasopressin (AVP) or oxytocin (OT), two nine-amino-acid neuropeptides with diverse forebrain projections, could mediate the development of selective aggression and affiliation. This hypothesis was based on the following observations: (1) monogamous and polygamous voles differ specifically in the distribution of forebrain AVP and OT receptors; (2) AVP innervation in the prairie vole brain is sexually dimorphic and important for paternal behaviour; (3) central AVP pathways have been previously implicated in territorial displays and social memory; and (4) central OT pathways have been previously implicated in affiliative behaviours. We now demonstrate that central AVP is both necessary and sufficient for selective aggression and partner preference formation, two critical features of pair bonding in the monogamous prairie vole. PMID- 8413609 TI - African and North American populations of Drosophila melanogaster are very different at the DNA level. AB - Understanding genetic evolution within species requires an accurate description of variation within and between populations and the ability to distinguish between the potential causes of an observed distribution of variation. In the cosmopolitan species Drosophila melanogaster, previous studies suggested that gene flow within and between continents is extensive and that most of the nuclear gene variation is found within, rather than among, populations. Here we present evidence that a population from Zimbabwe is more than twice as variable as those from the United States of America at the DNA sequence level, that most variants are not shared between the two geographic regions, and that there are nearly fixed differences between the Zimbabwe and USA samples in genomic regions experiencing low recombination rates. It appears that there is an unappreciated degree of population structure in D. melanogaster and that equilibrium models of molecular evolution are inappropriate for this species. PMID- 8413610 TI - Macaque V1 neurons can signal 'illusory' contours. AB - We describe here a new view of primary visual cortex (V1) based on measurements of neural responses in V1 to patterns called 'illusory contours' (Fig. 1a, b). Detection of an object's boundary contours is a fundamental visual task. Boundary contours are defined by discontinuities not only in luminance and colour, but also in texture, disparity and motion. Two theoretical approaches can account for illusory contour perception. The cognitive approach emphasizes top-down processes. An alternative emphasizes bottom-up processing. This latter view is supported by (1) stimulus constraints for illusory contour perception and (2) the discovery by von der Heydt and Peterhans of neurons in extrastriate visual area V2 (but not in V1) of macaque monkeys that respond to illusory contours. Using stimuli different from those used previously, we found illusory contour responses in about half the neurons studied in V1 of macaque monkeys. Therefore, there are neurons as early as V1 with the computational power to detect illusory contours and to help distinguish figure from ground. PMID- 8413611 TI - Protein tyrosine kinase p56lck controls allelic exclusion of T-cell receptor beta chain genes. AB - During T-cell development, site-specific DNA rearrangements mediating assembly of beta- and alpha-chain genes of the T-cell receptor (TCR) are developmentally ordered. In particular, assembly and expression of a complete beta-chain gene blocks further rearrangements at the beta-locus (a process referred to as allelic exclusion) and drives the generation and expansion of CD4+8+ cells. Although the mechanism used by TCR beta chains to deliver such signals is unknown, studies in transgenic animals have suggested that the lymphocyte-specific protein tyrosine kinase p56lck may impinge on a similar signalling pathway. The hypothesis that TCR beta chains deliver intracellular signals via p56lck makes an explicit prediction: that interference with p56lck function will mitigate the effects of a simultaneously expressed TCR beta chain. Here we confirm this prediction through examination of allelic exclusion in mice expressing both a functional TCR beta chain transgene and a catalytically inactive form of p56lck. PMID- 8413612 TI - An activated Notch receptor blocks cell-fate commitment in the developing Drosophila eye. AB - The Notch locus of Drosophila melanogaster encodes a 2,703-amino-acid transmembrane protein required for a variety of developmental processes, including neurogenesis, oogenesis and ommatidial assembly. The Notch protein contains a large extracellular domain of 36 epidermal growth factor-like repeats as well as three Notch/Lin-12 repeats and an intracellular domain with 6 Cdc10/ankyrin repeats, motifs that are highly conserved in several vertebrate Notch homologues. Truncation of the extracellular domain of the Drosophila Notch protein produces an activated receptor, as judged by its ability to cause phenotypes similar to gain-of-function alleles of duplications of the Notch locus. Equivalent truncations of vertebrate Notch-related proteins have been associated with malignant neoplasma and other developmental abnormalities. We present here an analysis of activated Notch function at single-cell resolution in the Drosophila compound eye. We find that overexpression of full-length Notch in defined cell types has no apparent effects but that overexpression of activated Notch in the same cells transiently blocks their proper cell-fate commitment, causing them either to adopt incorrect cell fates or to differentiate incompletely. Moreover, an activated Notch protein lacking the transmembrane domain is translocated to the nucleus, raising the possibility that Notch may participate directly in nuclear events. PMID- 8413613 TI - Sphingosine-1-phosphate as second messenger in cell proliferation induced by PDGF and FCS mitogens. AB - Growth signalling networks that use glycerophospholipid metabolites as second messengers have been well characterized, but less is known of the second messengers derived from sphingolipids, another major class of membrane lipids. A tantalizing link between sphingolipids and cellular proliferation has emerged from the discovery that the sphingolipid metabolites sphingosine and sphingosine 1-phosphate stimulate growth of quiescent Swiss 3T3 fibroblasts by a pathway that is independent of protein kinase C. Sphingosine-1-phosphate is rapidly produced from sphingosine and may mediate its biological effects. Furthermore, sphingosine 1-phosphate triggers the dual signal transduction pathways of calcium mobilization and activation of phospholipase D, prominent events in the control of cellular proliferation. Here we report that activation of sphingosine kinase and the formation of sphingosine-1-phosphate are important in the signal transduction pathways activated by the potent mitogens platelet-derived growth factor (PDGF) and fetal calf serum (FCS). PMID- 8413614 TI - Cell-cycle control of a large-conductance K+ channel in mouse early embryos. AB - There have been few investigations into the role of ion channels in mammalian early embryonic development, despite studies showing that changes in ion channel activity accompany the early embryonic development of non-mammalian species and the proliferation of mammalian cells. Here we report that a large-conductance, voltage-activated K+ channel is active in unfertilized mouse oocytes but is rarely observed in later embryos. The channel activity is linked to the cell cycle, being active throughout M and G1 phases, and switching off during the G1 to-S transition. These changes in channel activity are accompanied by corresponding shifts in membrane potential. Inactivation of the channel during S/G2 can be prevented by exposing the oocytes to dibutyryl cyclic AMP or forskolin, an activator of adenylyl cyclase. Inhibition of protein synthesis with puromycin did not prevent inactivation of the channel at the end of G1 or its subsequent reactivation at the end of G2, indicating that the channel activity is not regulated by mitosis-promoting factor or cyclins. PMID- 8413615 TI - Residues in the TATA-binding protein required to mediate a transcriptional response to retinoic acid in EC cells. AB - The eukaryotic TATA-binding protein TBP, which is required for transcription by RNA polymerase II, is tightly associated with a particular set of factors in the TFIID complex, and as such provides a target for transcriptional regulation exerted by upstream factors. An embryonic carcinoma (EC) cell-specific activity like that of the viral factor E1A has been implicated in the mediation of transactivation from the retinoic acid receptor to human TBP, but yeast TBP cannot perform this function. Using TBP mutants with an altered TATA-box-binding specificity, we show here that yeast TBP can mediate transcriptional activation in mammalian cells and that its inability to convey retinoic acid-dependent transactivation in EC cells is due to specific residues in its core region. These residues preclude a functional association with the cellular E1A-like activity. TBP is thus a target for retinoic acid-dependent transactivation in EC cells by providing a surface for interaction with the EC cell-specific E1A-like activity. PMID- 8413616 TI - Senior German health officials sacked. PMID- 8413617 TI - Watson to take the helm at Soros institute. PMID- 8413618 TI - UK plans library of DNA profiles. PMID- 8413619 TI - France urged to change ethics rule. PMID- 8413620 TI - Nobel goes to discoverers of 'split genes'. PMID- 8413621 TI - Allain defended. PMID- 8413622 TI - G-protein-coupled receptors. Turned on to ill effect. PMID- 8413623 TI - Cell physiology. Fatty feedback and fluidity. PMID- 8413624 TI - Music and spatial task performance. PMID- 8413625 TI - MyoD and c-fos expression. PMID- 8413626 TI - The daf-4 gene encodes a bone morphogenetic protein receptor controlling C. elegans dauer larva development. AB - The bone morphogenetic protein (BMP) family is a conserved group of signalling molecules within the transforming growth factor-beta (TGF-beta) superfamily. This group, including the Drosophila decapentaplegic (dpp) protein and the mammalian BMPs, mediates cellular interactions and tissue differentiation during development. Here we show that a homologue of human BMPs controls a developmental switch in the life cycle of the free-living soil nematode Caenorhabditis elegans. Starvation and overcrowding induce C. elegans to form a developmentally arrested, third-stage dauer larva. The daf-4 gene, which acts to inhibit dauer larva formation and promote growth, encodes a receptor protein kinase similar to the daf-1, activin and TGF-beta receptor serine/threonine kinases. When expressed in monkey COS cells, the daf-4 receptor binds human BMP-2 and BMP-4. The daf-4 receptor is the first to be identified for any growth factor in the BMP family. PMID- 8413627 TI - Somatic mutations in the thyrotropin receptor gene cause hyperfunctioning thyroid adenomas. AB - The pituitary hormone thyrotropin stimulates the function, expression of differentiation and growth of thyrocytes by cyclic AMP-dependent mechanisms. Tissue hyperplasia and hyperthyroidism are therefore expected to result when activation of the adenylyl cyclase-cAMP cascade is unregulated. This is observed in several situations, including when somatic mutations impair the GTPase activity of the G protein Gsa (ref 6, 7). Such a mechanism is probably responsible for the development of a minority of monoclonal hyperfunctioning thyroid adenomas. Here we identify somatic mutations in the carboxy-terminal portion of the third cytoplasmic loop of the thyrotropin receptor in three out of eleven hyperfunctioning thyroid adenomas. These mutations are restricted to tumour tissue and involve two different residues (aspartic acid at position 619 to glycine in two cases, and alanine at position 623 to isoleucine in one case). The mutant receptors confer constitutive activation of adenylyl cyclase when tested by transfection in COS cells. This shows that G-protein-coupled receptors are susceptible to constitutive activation by spontaneous somatic mutations and may thus behave as proto-oncogenes. PMID- 8413628 TI - Prevention of lung reperfusion injury in rabbits by a monoclonal antibody against interleukin-8. AB - Re-establishing blood flow to ischaemic tissues causes greater injury than that induced during the ischaemic period. This type of tissue injury, reperfusion injury, is involved in frostbite, multiple organ failure after hypovolaemia and in myocardial infarction. Depletion of neutrophils alleviates reperfusion injury, implying a causal role of neutrophil infiltration. Among members of the recently discovered family of chemotactic cytokines (chemokines), interleukin-8 (IL-8) is a major neutrophil chemotactic and activating factor produced by various types of human cells. We investigated its pathophysiological role in a rabbit model of a lung reperfusion injury. Reperfusion of ischaemic lung caused neutrophil infiltration and destruction of pulmonary structure, as well as local production of IL-8. Furthermore, the administration of a neutralizing monoclonal antibody against IL-8 prevented neutrophil infiltration and tissue injury, proving a causal role of locally produced IL-8 in this model. PMID- 8413629 TI - Coupling of the Na+/Ca2+ exchanger, Na+/K+ pump and sarcoplasmic reticulum in smooth muscle. AB - The Na+/Ca2+ exchanger, driven by a transmembrane Na+ gradient, plays a key role in regulating Ca2+ concentration in many cells. Although the exchanger influences Ca2+ concentration, its activity in smooth muscle appears to be closely coupled to Ca2+ availability from intracellular stores. This linkage might result if the exchanger were positioned close to Ca2+ storage sites within the sarcoplasmic reticulum. To test this hypothesis we have developed methods to assess the relative three-dimensional distribution of proteins involved in Na+/K+ pumping, Na+/Ca2+ exchange, Ca2+ storage within the sarcoplasmic reticulum, and attachment of contractile filaments to the membrane in smooth muscle. Here we report that the Na+/Ca2+ exchanger is largely co-distributed with the Na+/K+ pump on unique regions of the plasma membrane in register with, and close to, calsequestrin containing regions of the sarcoplasmic reticulum in sites distinct from the sites where contractile filaments attach to the membrane. This molecular organization suggests that the plasma membrane is divided into at least two functional domains, and appear to provide a mechanism for the strong linkage seen in smooth muscle between Na+/K+ pumping and Na+/Ca2+ exchange, and between Na+/Ca2+ exchange and Ca2+ release from the sarcoplasmic reticulum. PMID- 8413630 TI - The GTP-binding protein Ran/TC4 is required for protein import into the nucleus. AB - Two cytosolic fractions (A and B) from Xenopus oocytes are sufficient to support protein import into the nuclei of digitonin-permeabilized cells. Fraction A recognizes the nuclear localization sequence (NLS) and binds the import substrate to the nuclear envelope, whereas fraction B mediates the subsequent passage of the bound substrate into the nucleus. Here we report that two interacting components are required for full fraction-B activity, purify one of these components to homogeneity, and show that it is the highly abundant GTP-binding protein Ran (Ras-related nuclear protein)/TC4. PMID- 8413631 TI - ATP-induced protein-Hsp70 complex dissociation requires K+ but not ATP hydrolysis. AB - The molecular chaperone proteins, particularly Hsp60 and Hsp70, have been implicated in essential cell functions under both normal and stress conditions (reviewed in refs 1-5). Members of the family of heat-shock proteins of M(r) 70K, Hsp70, bind to unfolded proteins and short peptides. Addition of Mg-ATP results in the dissociation of the substrate polypeptides from the chaperone, but as ATP gamma S (an ATP analogue that is only slowly hydrolysable) cannot substitute for ATP in this reaction, it has been concluded that ATP hydrolysis is necessary to dissociate Hsp70-substrate protein complexes. By independently measuring the rates of ATP hydrolysis and substrate protein dissociation, we show here that Mg ATP binding but not Mg-ATP hydrolysis is essential for substrate dissociation. We also show that there is an absolute requirement for K+ for the effect of Mg-ATP: only the combination of K+ and Mg-ATP will cause the conformational change in Hsp70 that is necessary for substrate dissociation. Moreover, in the absence of K+, Mg-ATP favours complex formation. We consider these results in terms of a G protein-like model. PMID- 8413632 TI - Quantitative detection of HIV-1 drug resistance mutations by automated DNA sequencing. AB - A comparison has been made between manual and automated DNA sequencing procedures to evaluate the ability to distinguish mixtures of wild-type and mutant sequences. Quantitative detection of such mixtures of HIV-1 drug resistance mutations was best achieved using an automated system that uses fluorescent labelled sequencing primers. This procedure has a wide range of applications in clinical research, including heterozygote analysis. Software that automatically reports mixed-base positions is presented. PMID- 8413633 TI - A window of opportunity. AB - Still in shock from sepsis, biotechnology now has to face healthcare Clinton style. But research in biotechnology is on the increase as its products surge towards the markets. PMID- 8413634 TI - Modest growth for NIH. PMID- 8413635 TI - Biotechnology rules: Spain falls into line. PMID- 8413636 TI - German changes face opposition. PMID- 8413637 TI - UK report criticizes EC directives. PMID- 8413638 TI - Nobel rewards two laboratory revolutions. PMID- 8413639 TI - Bid to launch 'European PhD'. PMID- 8413640 TI - French gene laboratory gets a new lease of life. PMID- 8413641 TI - Minister disbands German health bureau. PMID- 8413642 TI - US 'should re-create national bioethics forum'. PMID- 8413643 TI - Common inheritance. PMID- 8413644 TI - Electronic journals are already here. PMID- 8413645 TI - Localized learning in insects. PMID- 8413646 TI - Molecular motors. One giant step for kinesin. PMID- 8413647 TI - Recovery of antediluvian DNA. PMID- 8413648 TI - Child leukaemia after Chernobyl. PMID- 8413649 TI - Sexual orientation. PMID- 8413650 TI - Direct observation of kinesin stepping by optical trapping interferometry. AB - Do biological motors move with regular steps? To address this question, we constructed instrumentation with the spatial and temporal sensitivity to resolve movement on a molecular scale. We deposited silica beads carrying single molecules of the motor protein kinesin on microtubules using optical tweezers and analysed their motion under controlled loads by interferometry. We find that kinesin moves with 8-nm steps. PMID- 8413651 TI - Cope's rule, the island rule and the scaling of mammalian population density. AB - Cope's rule--the generalization that animal taxa tend to evolve toward larger body size--suggests that there are widespread net selective advantages to being large. Size-abundance relationships within bird and desert rodent guilds show that larger species usually do control more energy locally, and thus maintain larger populations than expected for their body size, implying that larger individuals are relatively better at obtaining and using local resources. But we report here results that show that this is not generally the case among mammal species. Within dietary groups containing only small species, larger species usually do better, but within those that contain the largest mammals, small species tend to control more energy. This suggests that in mammals there is an optimum body size for energy acquisition at about 1 kg. Thus, net adaptive advantages of large individuals for resource control cannot be used as a general explanation for evolutionary size increase in mammals, although other proposed explanations for Cope's rule are unaffected. Instead, these results suggest a partial explanation for another widespread ecotypic pattern, the 'island rule': that on islands, small mammal species evolve to larger size and large species to smaller size. If on an island a species' usual competitors and predators are absent, it should often tend to evolve toward the optimum body size, and the adaptive advantages of doing so would be greatest for populations starting at body-size extremes. PMID- 8413652 TI - Visual pattern recognition in Drosophila involves retinotopic matching. AB - Honeybees remember the shapes of flowers and are guided by visual landmarks on their foraging trips. How insects recognize visual patterns is poorly understood. Experiments suggest that they try to match retinotopically the incoming visual pattern with a previously stored memory image. But bees can be conditioned to individual pattern parameters such as orientation of contours, colour or size. These and other results are difficult to reconcile with simple template matching. In such investigations, freely moving animals are observed; their behaviour and visual input, therefore, are not well known. Mostly, processing strategies are inferred from stimulus design. We have studied visual pattern recognition with tethered flies (Drosophila melanogaster) in a flight simulator and report here that flies store visual images at, or together with, fixed retinal positions and can retrieve them from there only. Position invariance, an acknowledged property of human pattern recognition, may not exist as a primary mechanism in insects. PMID- 8413653 TI - Prefrontal neuronal activity in rhesus monkeys performing a delayed anti-saccade task. AB - Patients with damage to the dorsolateral prefrontal cortex are impaired on cognitive tasks such as the Wisconsin Card Sort Test, the Stroop Test and an anti saccade paradigm, in which sensory-guided habitual responses must be suppressed in favour of conceptually or memory-guided responses. We report here recordings from prefrontal neurons in rhesus monkeys trained to perform a delayed anti saccade task based on tests that have been used with humans. Activity in the same prefrontal neurons was recorded across conditions when saccades were made toward a remembered target, and also when this prepotent response was suppressed and a saccade in the opposite direction required. Our findings show that most prefrontal neurons code the location of the visual stimulus in working memory, and that this memory can be engaged to suppress as well as prescribe a response. These results establish, in a subset of prefrontal neurons, the iconic nature of the memory code, and suggest a role for visual memory in response suppression. PMID- 8413654 TI - MIF is a pituitary-derived cytokine that potentiates lethal endotoxaemia. AB - Cytokines are critical in the often fatal cascade of events that cause septic shock. One regulatory system that is likely to be important in controlling inflammatory responses is the neuroendocrine axis. The pituitary, for example, is ideally situated to integrate central and peripheral stimuli, and initiates the increase in systemic glucocorticoids that accompanies host stress responses. To assess further the contribution of the pituitary to systemic inflammatory processes, we examined the secretory profile of cultured pituitary cells and whole pituitaries in vivo after stimulation with bacterial lipopolysaccharide (LPS). Here we identify macrophage migration inhibitory factor (MIF) as a major secreted protein release by anterior pituitary cells in response to LPS stimulation. Serum analysis of control, hypophysectomized and T-cell-deficient (nude) mice suggests that pituitary-derived MIF contributes to circulating MIF present in the post-acute phase of endotoxaemia. Recombinant murine MIF greatly enhances lethality when co-injected with LPS and anti-MIF antibody confers full protection against lethal endotoxaemia. We conclude that MIF plays a central role in the toxic response to endotoxaemia and possibly septic shock. PMID- 8413655 TI - Pathophysiological role of endothelin revealed by the first orally active endothelin receptor antagonist. AB - Since its discovery, endothelin-1 has attracted considerable scientific interest because of its extremely potent and long-lasting vasoconstrictor effect and its binding to G-protein-coupled receptors. Plasma concentrations of endothelin-1 are low and its release by endothelial cells is polarized towards the basolateral side, suggesting that it is a paracrine factor and not a hormone. Consequently, the effect of injected endothelin-1 may not reflect the effect of endogenous endothelin-1. In contrast, blockade of the action of endogenous endothelin-1 using receptor antagonists should be a valuable means of investigating its physiological and pathological effects. We report here evidence for the pathophysiological role of endothelin-1 as brought by the first synthetic orally active nonpeptide antagonist of endothelin receptors, Ro 46-2005. PMID- 8413656 TI - Protein composition determines the anti-atherogenic properties of HDL in transgenic mice. AB - High-density lipoprotein (HDL) contains two major proteins, apolipoprotein A-I (apoA-I) and apolipoprotein A-II (apoA-II), comprising about 70% and 20% of the total HDL protein mass, respectively. HDL exists in human plasma in two main forms, one containing apoA-I with apoA-II (AI/AII-HDL) and another containing apoA-I without apoA-II (AI-HDL). A strong inverse relationship exists between total plasma HDL concentration and atherosclerosis, but the results of studies examining the relationship between AI-HDL and AI/AII-HDL and atherosclerosis have been conflicting. To determine whether these two HDL populations have different effects on atherogenesis, human apoA-I (AI) and human apoA-I and apoA-II (AI/AII) transgenic mice were produced in an atherosclerosis-susceptible strain. Following an atherogenic diet, despite similar total cholesterol and HDL cholesterol concentrations, the area of atherogenic lesions in the AI/AII mice was 15-fold greater than in the AI animals. These studies show that the protein composition of HDL significantly affects its role in atherogenesis and that AI-HDL is more antiatherogenic than AI/AII-HDL. PMID- 8413657 TI - South African cyclotron faces uncertain future. PMID- 8413658 TI - Childhood leukaemias remain unexplained. PMID- 8413659 TI - Cloning of human embryos draws fire from critics. PMID- 8413660 TI - German university faces ban on animal tests in class. PMID- 8413661 TI - The pathway to signal achievement. PMID- 8413662 TI - Molecular motors. Keeping out the rain. PMID- 8413663 TI - Palaeoanthropology. Rift on the record. PMID- 8413664 TI - Immunological modulation and evasion by helminth parasites in human populations. AB - Helminth parasites are highly prevalent in human communities in developing countries. In an endemic area an infected individual may harbour parasitic worms for most of his or her life, and the ability of these infections to survive immunological attack has long been a puzzle. But new techniques are starting to expose the diverse mechanisms by which these agents modulate or evade their hosts' defences, creating a dynamic interaction between the human immune system and the parasite population. PMID- 8413665 TI - The structure of crystalline profilin-beta-actin. AB - The three-dimensional structure of bovine profilin-beta-actin has been solved to 2.55 A resolution by X-ray crystallography. There are several significant local changes in the structure of beta-actin compared with alpha-actin as well as an overall 5 degrees rotation between its two major domains. Actin molecules in the crystal are organized into ribbons through intermolecular contacts like those found in oligomeric protein assemblies. Profilin forms two extensive contacts with the actin ribbon, one of which appears to correspond to the solution contact in vitro. PMID- 8413666 TI - Oldest Homo and Pliocene biogeography of the Malawi Rift. AB - The Malawi Rift and Pliocene palaeofaunas, which include a hominid mandible attributed to Homo rudolfensis, provide a biogeographical link between the better known Plio-Pleistocene faunal records of East and Southern Africa. The Malawi Rift is in a latitudinal position suitable for recording any hominid and faunal dispersion towards the Equator that was brought on by increased aridity of the Late Pliocene African landscape. The evidence suggests that Pliocene hominids originated in the eastern African tropical domain and dispersed to southern Africa only during more favourable ecological circumstances. PMID- 8413667 TI - Reverse transduction measured in the isolated cochlea by laser Michelson interferometry. AB - It is thought that the sensitivity of mammalian hearing depends on amplification of the incoming sound within the cochlea by a select population of sensory cells, the outer hair cells. It has been suggested that these cells sense displacements and feedback forces which enhance the basilar membrane motion by reducing the inherent damping of the cochlear partition. In support of this hypothesis, outer hair cells show membrane-potential-induced length changes at acoustic rates. This process has been termed 'reverse transduction'. For amplification, the forces should be large enough to move the basilar membrane. Using a displacement sensitive interferometer, we tested this hypothesis in an isolated cochlea while stimulating the outer hair cells with current passed across the partition. We show here that the cochlear partition distorts under the action of electrically driven hair cell length changes and produces place-specific vibration of the basilar membrane of a magnitude comparable to that observed near auditory threshold (about 1 nm). Such measurements supply direct evidence that cochlear amplification arises from the properties of the outer hair cell population. PMID- 8413668 TI - Inhibition of contractile vacuole function in vivo by antibodies against myosin I. AB - Myosin-I is thought to supply the force for movement of cell membranes relative to actin filaments (reviewed in refs 1, 2), but confirmation of this hypothesis has been difficult because of the presence of multiple isoforms of myosin-I and other unconventional myosins in most cells. We report here the first evidence that a myosin-I isoform is essential for a specific class of intracellular membrane movements in vivo. In Acanthamoeba, the contractile vacuole is an autonomous structure which fuses with the plasma membrane to control the water content of the cell. Because myosin-IC is the only myosin-I isoform concentrated in the contractile vacuole complex, and a protein antigenically related to myosin IC is located on or near the Dictyostelium (slime mould) contractile vacuole, we thought this organelle might provide the best opportunity to demonstrate a relationship between myosin-I and membrane motility. Antibodies that inhibit the activity of Acanthamoeba myosin-IC in vitro interfere with expulsion of excess water by the contractile vacuole in vivo, leading to overfilling of this organelle and cell lysis. Myosin-IC may generate the force required to contract the vacuole and may also be involved in transfer of water to the contractile vacuole during refilling. PMID- 8413669 TI - Cloning and characterization of the vasopressin-regulated urea transporter. AB - Urea is the principal end product of nitrogen metabolism in mammals. Movement of urea across cell membranes was originally thought to occur by lipid-phase permeation, but recent studies have revealed the existence of specialized transporters with a low affinity for urea (Km > 200 mM)2. Here we report the isolation of a complementary DNA from rabbit renal medulla that encodes a 397 amino-acid membrane glycoprotein, UT2, with the functional characteristics of the vasopressin-sensitive urea transporter previously described in in vitro-perfused inner medullary collecting ducts. UT2 is not homologous to any known protein and displays a unique pattern of hydrophobicity. Because of the central role of this transporter in fluid balance and nitrogen metabolism, the study of this protein will provide important insights into the urinary concentrating mechanism and nitrogen balance. PMID- 8413670 TI - Identification of a proline residue as a transduction element involved in voltage gating of gap junctions. AB - Gap junction channels are structurally distinct from other ion channels in that they comprise two hemichannels which interact head-to-head to form an aqueous channel between cells. Intercellular voltage differences together with increased intracellular concentrations of H+ and Ca2+ cause closure of these normally patent channels. The relative sensitivity to voltage varies with the subunit (connexin) composition of the channels. The third of four transmembrane-spanning regions (M3) in connexins has been proposed to form the channel lining, and a global 'tilting' of the hemichannel subunits has been correlated with channel closure. But specific components involved in transduction of channel gating events have not been identified in either gap junctions or other ion channel classes (however, see model in ref. 5). We have examined a strictly conserved proline centrally located in M2 of connexin proteins. Mutation of this proline (Pro 87) in connexin 26 causes a reversal in the voltage-gating response when the mutant hemichannel is paired with wild-type connexin 26 in the Xenopus oocyte system. This suggests that the unique properties associated with this residue are critical to the transduction of voltage gating in these channels. PMID- 8413671 TI - The protein IsK is a dual activator of K+ and Cl- channels. AB - The protein IsK (M(r) 14,500) is present in epithelial cells, heart, uterus and lymphocytes and induces slowly activating K+ currents when expressed in Xenopus oocytes. The finding that mutations of its single transmembrane segment altered channel gating or selectivity has suggested that IsK is a channel-forming protein. But IsK does not exhibit the K+ channel hallmarks (a conserved K+ selective pore (H5) flanked by either six or two membrane-spanning regions). Here we report that IsK expression in Xenopus oocytes also induces a Cl- selective current very similar to the Cl- current produced by phospholemman expression and with biophysical, pharmacological and regulation characteristics very different from those of the IsK-induced K+ channel activity. IsK mutagenesis identifies amino- and carboxy-terminal domains as critical for the induction of Cl- and K+ channel activities, respectively. Our data lead to a model in which the IsK protein (now called IsK, Cl) acts as a potent activator of endogenous and otherwise silent K+ or Cl- channels. PMID- 8413672 TI - Human xeroderma pigmentosum group D gene encodes a DNA helicase. AB - Xeroderma pigmentosum (XP), a genetically heterogeneous human disease, results from a defect in nucleotide excision repair of ultraviolet-damaged DNA. XP patients are extremely sensitive to sunlight and suffer from a high incidence of skin cancers. Cell fusion studies have identified seven XP complementation groups, A-G. Group D is of particular interest as mutations in this gene can also cause Cockayne's syndrome and trichothiodystrophy. The XPD gene was initially named ERCC2 (excision repair cross complementing) as it was cloned using human DNA to complement the ultraviolet sensitivity of a rodent cell line. We have purified the XPD protein to near homogeneity and show that it possesses single stranded DNA-dependent ATPase and DNA helicase activities. We tested whether XPD can substitute for its yeast counterpart RAD3, which is essential for excision repair and for cell viability. Expression of the XPD gene in Saccharomyces cerevisiae can complement the lethality defect of a mutation in the RAD3 gene, suggesting that XPD is an essential gene in humans. PMID- 8413673 TI - Phosphorylated CREB binds specifically to the nuclear protein CBP. AB - Cyclic AMP-regulated gene expression frequently involves a DNA element known as the cAMP-regulated enhancer (CRE). Many transcription factors bind to this element, including the protein CREB, which is activated as a result of phosphorylation by protein kinase A. This modification stimulates interaction with one or more of the general transcription factors or, alternatively, allows recruitment of a co-activator. Here we report that CREB phosphorylated by protein kinase A binds specifically to a nuclear protein of M(r) 265K which we term CBP (for CREB-binding protein). Fusion of a heterologous DNA-binding domain to the amino terminus of CBP enables the chimaeric protein to function as a protein kinase A-regulated transcriptional activator. We propose that CBP may participate in cAMP-regulated gene expression by interacting with the activated phosphorylated form of CREB. PMID- 8413674 TI - Molecular basis of crossreactivity and the limits of antibody-antigen complementarity. AB - Two major unanswered questions concerning the specificity of antibodies are: how do structurally different antigens bind with high affinity to the same antibody, and what are the limits of the antibody combining site complementarity and flexibility that contribute to such crossreactivity? We report here a comparative analysis of the X-ray structures of five conformationally different steroids in complex with the Fab' fragment of an anti-progesterone antibody DB3 at 2.7 A. This antibody is unable to complement completely the shape of the hydrophobic antigen so that crossreactivity occurs with other ligands without major structural rearrangements of the binding site. Antigen specificity can be explained through conserved interactions of DB3 with the steroid D-ring, whereas some of the crossreactivity is realized through different binding orientations of the steroid skeleton that place the A-ring into alternative pockets on the antibody surface. The restricted gene usage of the VGAM3.8 family in the generation of anti-progesterone monoclonal antibodies may be explained by the specific interaction of VH hallmark residues with the steroid D-ring. This first detailed structure of steroid interactions with a protein could be applied to the understanding of general mechanisms of steroid recognition as well as in the design of specific binding sites for small hydrophobic ligands. PMID- 8413675 TI - [Sugar analog inhibitors for glycosidases, tools for the elucidation of enzymatic hydrolysis of glycosides]. AB - Sugar derivatives with a basic group on C-1 (glycosylamines, 5-amino-5 deoxypyranoses, and 1,5-iminohexitols) are bound by most glycosidases 10(2)- to 10(5)-fold more tightly than their nonbasic counterparts. This high affinity and an up to 10(5)-fold better inhibition relative to hexoses by hexono-delta lactones and lactams point to a catalytic mechanism characterized by a transition state with a partial positive charge and planar geometry at the anomeric carbon of the substrate. Protonation of the glycosidic oxygen atom and stabilization of the positive charge by a carboxylate group strongly shielded from the aqueous environment lower the free energy of activation to an extent which causes an up to 10(14)-fold rate acceleration relative to the nonenzymatic hydrolysis of glycosides. PMID- 8413676 TI - [Patent protection of biotechnological developments]. AB - Patents can be granted with respect to microbiological processes and products therefrom. In principle it is necessary to deposit a culture of the microorganisms at a recognized depository institution. The deposited culture is available upon request to any person from the date of publication of the patent application. Consequently, there is a great risk that the applicant's product is used by other people. PMID- 8413677 TI - [The many faces of the acid-fast rod]. PMID- 8413678 TI - [The right to information in medical practice]. PMID- 8413679 TI - [Dissociation, hypnosis and multiple personality disorders]. PMID- 8413680 TI - [Dissociative dis, especially multiple personality disorder]. PMID- 8413681 TI - [Cell and disease. II. The cell nucleus in pathology]. PMID- 8413682 TI - [Routine intravenous pyelography not diagnostically useful in men with lower urinary tract infections]. AB - OBJECTIVE: Evaluation of intravenous urography (IVU) in men with a lower urinary tract infection. SETTING: Leyenburg Hospital, the Hague, the Netherlands. DESIGN: Retrospective. METHODS: In 28 male patients with a diagnosed lower urinary tract infection, an IVU was made in order to find causative urogenital tract abnormalities. We determined whether the IVU provided additional diagnostic information. RESULTS: Only I IVU (4%) of the 28 showed abnormalities (a bladder stone in a patient with epididymitis). The finding was not statistically significant (Fisher's exact test). CONCLUSION: In men with a symptomatic lower urinary tract infection showing symptoms such as frequent and burning micturition, whether or not complicated or not by epididymitis, an IVU gives no additional diagnostic information. PMID- 8413683 TI - [Laparoscopic cystectomy of the ovary; results of a new technique]. AB - The results of a prospective study of laparoscopic management of ovarian cysts are discussed. This technique of minimally invasive surgery was introduced at the department of Gynaecology of the Leiden University Medical Centre in 1991. Only premenopausal women with unilocular ovarian cysts smaller than 10 cm in diameter and with sonographically benign characteristics were included in this study. In a period of one year, 25 cystectomies and one ovariectomy were performed. 88% of the cysts were initially treated with hormone therapy. Mean age of the patients was 30 years (range: 21-54), average size of the cysts was 6 cm (range: 4-10) and mean operating time was 80 minutes (range: 35-120), there were no complications during operation or in the postoperative period. No laparotomies had to be performed. Three times (11.5%) pathological specimen examination revealed a corpus luteum cyst. The other 23 were non-functional cysts. No carcinoma was detected. The results of this minimally invasive surgery for removal of ovarian cysts are encouraging. Morbidity compared with the conventional cystectomy by laparotomy is low as appears from a shorter hospital stay and quicker recovery. Cystectomy of the ovary, compared with laparoscopic aspiration and fenestration, has a better therapeutic effect and the histological evaluation is more reliable. Persistent, unilocular ovarian cysts, which fulfill the criteria used in our study, are best treated by laparoscopic removal instead of laparotomy. PMID- 8413684 TI - [Should the physician be able to prove that he informed the patient satisfactorily?]. PMID- 8413686 TI - [Patient's rights in Europe]. PMID- 8413685 TI - [Patient information: observations, adjusted procedure, assessment]. PMID- 8413687 TI - [Sexual possibilities following total penis amputation]. PMID- 8413688 TI - [A Dutch family with familial Mediterranean fever]. PMID- 8413689 TI - [Endometriosis of the rectum]. PMID- 8413690 TI - [Dermatologically tested, can it be done in a better fashion?]. PMID- 8413691 TI - [Monoamine oxidase inhibitors: poisoning and interactions]. PMID- 8413692 TI - [Cell and disease. III. Apoptosis, the biological counterpart of mitosis]. PMID- 8413693 TI - [Home treatment of patients with cerebrovascular accidents]. AB - OBJECTIVE: In the Netherlands one-third of stroke patients are managed at home. This study describes the characteristics of these patients and the management by the GP from stroke onset till six months afterwards. METHOD: A community based cohort of 185 stroke patients were examined at fixed intervals during 6 months poststroke; mortality and functional status of patients managed at home (n = 73) were compared with those of patients admitted to hospital (n = 109); 3 patients were directly admitted to a nursing home; characteristics of the GPs' management were registered. RESULTS: Patients managed at home were somewhat older (p < 0.05), lived more often in a residential home for the elderly (p < 0.05) and were more frequently known with comorbidity such as diabetes mellitus and Alzheimer (p < 0.05). Their poststroke disabilities were less grave; mortality rate was much lower than in the hospitalised group (p < 0.05). Recovery showed the same pattern in both groups. In the acute phase the GP made frequent housecalls; after a few days there was a decline in contact frequency. The mean 'treatment duration' was 12 weeks. The amount of care provided by the GP did not correlate with stroke severity. CONCLUSION: In the acute phase, when the patients' clinical condition was still unstable and the patients' families needed support, the GPs made frequent house-calls. During the stroke episode the number of contacts and the length of the treatment duration were defined more by comorbidity and social factors than by the severity of stroke. Regarding the pattern of stroke recovery the mean treatment duration of 12 weeks appears rather short for optimal support and rehabilitation. PMID- 8413694 TI - [The treatment of myocardial infarction during the hospital stage and shortly thereafter: now and 10 years ago]. AB - OBJECTIVE: Comparison between the early and the late eighties of the application of thrombolysis and revascularisation in the acute phase of a myocardial infarction. LOCATION: University Hospital, Rotterdam. DESIGN: Prospective with historical comparison. METHODS: All patients admitted between May 1987 and May 1990 with a myocardial infarction and aged up to 71 years were included (n = 430). Numbers of procedures and survival during the following year were compared with data of patients admitted from 1981 to the end of 1983 (n = 706). RESULTS: In 1981-1983 thrombolytics were administered to 9% of the patients, in 1987-1990 to 40%. Revascularisation procedures during the next year were performed in 17% and 50% of the patients respectively. Hospital mortality decreased from 14% to 10% (p < 0.05), one-year survival increased from 75% to 83% (p < 0.05). For patients from 1987-1990 one-year survival was higher after thrombolysis treatment: 90% versus 78% without (p < 0.01), and after revascularisation: 94% versus 87% without (p < 0.01). CONCLUSION: Compared with 1981-1983 the treatment is at present more directed towards reperfusion and revascularisation of the ischaemic myocardium, resulting in invasive treatment in 50% of the patients now as opposed to 25% in the early eighties. The survival rate during the first year has improved. PMID- 8413695 TI - [Infants who cry a lot: incidence and management of this problem in family practice]. AB - OBJECTIVE: To determine how frequently general practitioners (GPs) are consulted because of excessively crying infants and how such infants are managed. SETTING: A stratified sample of 103 GP practices throughout The Netherlands. DESIGN: Descriptive. METHOD: 161 GPs from 103 practices for one year (in four groups, each for three months) recorded all their contacts with patients as a part of the National Study of Diseases and Items of Service in General Practice, conducted by the Netherlands Institute for Primary Health Care Research. An inventory was made of the infants who fulfilled the criteria of the 'colic syndrome' as to age, symptomatology and diagnosis. RESULTS: Of the infants seen by the GPs, 10% displayed symptoms of excessive crying, colic or restlessness. In over one-third of this group a colic-like diagnosis was made; of the children from zero to four months this proportion was two-thirds. The probability of a child in the age group of 0 to 4 months being seen by the GP with colic-like symptoms and diagnosis amounted to 7.1% (cumulative incidence). The majority of the GPs were certain of their (mostly somatic) diagnoses. The GPs were consulted more often about these children, apart from the colic problem, than about their contemporaries, the difference being statistically significant. In only 30% of the cases did the GPs prescribe medication or a diet, or made a referral. PMID- 8413696 TI - [Allergy to latex: an underestimated problem]. AB - Three patients who showed an allergic reaction to latex products are described. By means of history, skin tests, measurement of IgE antibodies against latex and challenge tests the diagnosis could be established. To prevent the spread of infectious diseases the use of latex gloves and condoms is increasing. Therefore, the incidence of allergic reactions to latex may also rise. Latex allergy should be considered when evaluating 'idiopathic' anaphylaxis during an operation. Moreover, latex allergy may be the cause of occupational rhinitis and asthma. PMID- 8413697 TI - [Peroperative anaphylactic reactions due to latex allergy]. AB - A 10-year-old girl with spina bifida underwent surgery to increase the capacity of her neurogenic bladder using an isolated loop of intestine. During the operation serious circulatory and respiratory problems occurred after the peritoneum was opened, when the ileum was manipulated using gloves containing latex. Latex allergy can lead to potentially fatal anaphylactic reactions. A survey is presented of the risk groups, together with measures recently advocated in the literature to detect latex allergy and to avoid major reactions. PMID- 8413698 TI - [Financing of health-related research. The pharmaceutical industry]. PMID- 8413699 TI - [Substantial decrease in number of hospital admissions for gonococcal infections in The Netherlands in 1981-1988]. PMID- 8413700 TI - [Korsakoff syndrome]. PMID- 8413701 TI - [The smart person will never sleep late; 'weekend headache' due to late and insufficient intake of caffeine]. PMID- 8413702 TI - [Is intravenous administration of immunoglobulins a first-choice method in autoimmune diseases?]. PMID- 8413703 TI - [Magnesium: a promising addition to the therapy in acute myocardial infarct]. PMID- 8413704 TI - [Cell and disease. IV. Cell pathological sequelae of ischemia and shock]. PMID- 8413705 TI - [Percutaneous gallbladder drainage a good treatment in patients with acute cholecystitis and and poor clinical status]. AB - OBJECTIVE: To evaluate the results of ultrasound-guided percutaneous cholecystostomy in patients with increased surgical risk. SETTING: University Hospital Rotterdam. DESIGN: Retrospective. MATERIAL AND METHOD: 24 patients with acute cholecystitis and an increased surgical risk were treated with ultrasound guided percutaneous cholecystostomy. Cholecystostomy was successfully performed in all patients. RESULTS: 23 patients showed clinical and biochemical improvement within one week. Minor complications were encountered in three cases (12%) which were treated conservatively. 16 patients suffered from calculous cholecystitis. One patient died after the cholecystostomy from an aspiration pneumonia. Cholecystectomy was performed at a later stage in six patients. One of these patients died six days after surgery from a myocardial infarction. Seven patients had percutaneous or endoscopic therapy for gallstones. In two cases no further therapy for gallstones was performed. Eight patients had acalculous cholecystitis. Four patients, suffering from cholangiocarcinoma and malignant obstruction of the cystic duct, subsequently underwent cholecystectomy (two patients) and percutaneous sclerotherapy of the gallbladder (two patients). Percutaneous cholecystostomy alone sufficed for the four remaining patients with acalculous cholecystitis. CONCLUSION: Percutaneous cholecystostomy for high-risk patients with acute cholecystitis is a low-invasive procedure with low morbidity and mortality. The catheter in the gallbladder offers diagnostic and therapeutic possibilities and does not interfere with cholecystectomy at a later stage. In some cases of acalculous cholecystitis drainage of the gallbladder is the only therapy needed. PMID- 8413706 TI - [Influenza epidemic in a nursing home caused by a virus not included in the vaccine]. AB - In the autumn of 1992 two-thirds of the population of a nursing home in Amsterdam was vaccinated against influenza. However, in March 1993 an outbreak of an influenza like illness occurred with a morbidity rate of 49% and a mortality rate of 10%. There was sufficient serological evidence to show that the vaccine as such had induced adequate immunity. As the causative agent an influenza A/H3N2 virus was identified. The failing activity of the vaccine in this instance was apparently caused by the absence of sufficient antigen similarity between the A/H3N2 vaccine component and the epidemic virus ('vaccine mismatch'). PMID- 8413707 TI - [A fatal influenza infection in a 10-year-old boy]. AB - A previously healthy 10-year-old boy died a few days after onset of septicaemia with non-specific clinical symptoms. Influenza B virus was isolated post mortem from pulmonary tissue. The histopathological findings did not indicate a virus disease. Specimens were taken for virus culture from other people in contact with the patient and affected with influenza-like illnesses. One other strain of influenza B virus was isolated. The strains could not be distinguished either serologically or genetically from other influenza B isolates of the season 1992/'93. PMID- 8413708 TI - [Intravenously-administered immunoglobulins as first-choice agent in juvenile dermatomyositis]. AB - Dermatomyositis is an acquired disease characterised by symmetric predominantly proximal muscle weakness of the arms and legs, and misery. It may be associated with myalgia and there is often a characteristic rash. The mainstay of therapy is corticosteroids. Recently efficacy of intravenous immunoglobulin (IVIg) in chronic refractory dermatomyositis was reported. Because corticosteroids can cause serious side effects, we treated a seven-year-old girl suffering from dermatomyositis with IVIg as initial therapy. After two courses of IVIg infusions at a dose of 0.4 g/kg/day for five consecutive days, the patient made a rapid and complete recovery. This case shows that IVIg may be effective as initial therapy in patients with dermatomyositis. Whether IVIg is really a better treatment than corticosteroids should be investigated in a randomised study. PMID- 8413709 TI - [Clinician, researcher and teacher Abraham Albert Hijmans van den Bergh (1869 1943)]. PMID- 8413710 TI - [Influenza in the 1992/93 season; vaccine composition for the 1993/94 season]. PMID- 8413711 TI - [Fertility-promoting drugs and ovarian carcinoma]. PMID- 8413712 TI - [Minor symptoms in family medicine; superficial skin burns]. PMID- 8413713 TI - [Depression sometimes the most important manifestation of hypothyroidism]. PMID- 8413714 TI - [Secondary poisoning (toxic substances in food chains): advice from the Public Health Council]. AB - The Dutch Health Council has issued a report on toxic contamination in food chains. The report discusses a model developed by the National Institute of Public Health and Environmental Protection and by the Tidal Waters Division of the Ministry of Transport, Public Works and Water Management. In this model, the accumulation of substances in aquatic and terrestrial food chains is described. The model uses the no-effect concentration for a fish-eating animal and the bioconcentration factor. Whether a toxic compound will accumulate in a food chain depends on the lipophilic character of the compound, its biodegradability, feeding habits of the particular animal involved, and the intrinsic toxicological susceptibility of the animal. The Health Council report emphasizes that the model has only limited value because the figures do not guarantee that higher animal species are sufficiently protected from toxic damage. Kinetic models such as physiology-based pharmacokinetic (PB-PK) models have not been used. These may increase the validity of extrapolation of animal experimental data to the human situation. PMID- 8413715 TI - [Chronic pancreatitis: the value of endoscopic therapy]. PMID- 8413716 TI - [What is the status of pancreas transplantation?]. PMID- 8413717 TI - [Cell and disease. V. Pathology of lysosomes]. PMID- 8413718 TI - [Good results of surgical treatment of idiopathic scoliosis using the Cotrel Dubousset technique]. AB - OBJECTIVE: To determine the results and complications of the Cotrel-Dubousset (CD) operation in idiopathic scoliosis. DESIGN: Retrospective. SETTING: Wilhelmina Children's Hospital, Utrecht, and University Hospital, Leiden. METHOD: Of the first 22 patients with idiopathic scoliosis operated on with CD instrumentation the results were determined: the Cobb angle, the percentage of rotational correction achieved, duration of the operation, amount of blood loss, complications and duration of the postoperative hospital stay. RESULTS: Compared with the Harrington technique, there were fewer complications resulting from instrumental failure (three in all), while the correction of the scoliosis was the same and in some cases even better. Advantages of the CD operation for the patient were rapid mobilization, short postoperative hospital stay and short duration of the after-treatment with use of a removable light plastic corset. CONCLUSION: The Cotrel-Dubousset operation appears to be a promising method for the treatment of patients with idiopathic scoliosis. PMID- 8413719 TI - [The treatment of liver metastases of colorectal tumors using intra-arterial fluorouracil via hepatic artery, administered with a completely implantable access system and a portable infusion pump]. AB - Thirty-seven patients with liver metastases from colorectal cancer were treated with hepatic arterial infusion of 5-FU. Before therapy an arterial access device (Port-A-Cath) was implanted. Treatment was given on an outpatient basis with a portable infusion pump. 5-FU was administered as a continuous infusion at a dose of 1000 mg/sq.m. per day during 5 or 6 days. Cycles were repeated every three weeks. In 12 (37.5%) of the 32 evaluable patients a partial remission was achieved. Median duration of remission was 11 months and median duration of survival was 16 months. Side effects of chemotherapy were usually mild. In 11 patients thrombosis of the hepatic artery was observed, occurring after a median period of 5 months after implantation of the access device. Further study is required to improve the treatment results in this disease with its unfavourable prognosis. PMID- 8413720 TI - [Reduction of the number of falls in hospitalized patients using a risk index and through preventive measures]. AB - OBJECTIVE: To investigate the effect of early identification of hospitalised patients with an increased risk of falling and of preventive measures on the frequency of falling. SETTING: Ten medical units (two neurology, five internal medicine, three surgery; 276 beds) in the St. Radboud Hospital in Nijmegen. METHODS: In a first case control study (5 months), a slightly modified falling risk index of Innes and Turman was validated. In a following (5.5 months) study period the index was computed for every hospitalised patient. Preventive measures were used in patients with a high score. The numbers of falls in both study periods were compared. All falls were carefully documented. RESULTS: High scores on the index were significantly associated with falls (p < 0.001). In both study periods (1 and 2) sensitivity (87% and 89%) and specificity (82% and 74%) of the index were high. A significant reduction in fall rate, 86 falls in period 1 versus 66 in period 2, was achieved, (corrected for number of patients and patient days). CONCLUSION: The modified index is a useful instrument for early identification of patients with a substantial risk of falling in the hospital. Early risk identification and preventive measures were successful in reducing the number of falls. PMID- 8413721 TI - [Acute pancreatitis in a patient with alpha 1-antitrypsin deficiency. Is there a causal relationship?]. AB - The clinical pattern and the pathophysiological mechanism of the hereditary disorder alpha 1-antitrypsin deficiency are outlined. It appeared that the patient described suffered not only from emphysema, cirrhosis of the liver and hepatocellular carcinoma, but also from acute haemorrhagic pancreatitis. The combination of acute pancreatitis and alpha 1-antitrypsin deficiency has not been described before. Screening of relatives is important because of possible treatment. PMID- 8413722 TI - [Mycetoma of the foot; a diseases from the tropics]. PMID- 8413723 TI - [2 children with discitis]. PMID- 8413724 TI - [Initial heparinization necessary in patients with thrombosis of the leg; comparison with acenocoumarol treatment]. PMID- 8413725 TI - [Motility disorders of stomach and proximal intestines in diabetes mellitus]. PMID- 8413726 TI - [Can the psychiatrist predict automobile driving ability]. PMID- 8413727 TI - [Disorders in the motility of stomach and small intestine in patients with insulin-dependent diabetes]. PMID- 8413728 TI - [Classification and diagnosis of personality disorders]. PMID- 8413729 TI - [Cell and disease. VI. Pathology of the cytoskeleton]. PMID- 8413730 TI - [Delay in hospitalization of patients with myocardial infarct]. AB - Timely treatment of patients with an evolving myocardial infarction improves the short and long term prognoses. Because of a wrong judgement of the situation by the patient, a significant other or by a general practitioner (GP), treatment may be delayed. To examine this delay 300 patients with myocardial infarction took part in a study between March 1990 and October 1991. After written consent was given, they were interviewed about the pre-hospital period. The significant others received a questionnaire about this period. Medical information was collected from the cardiologists. Fifty percent of all patients called for medical help within 30 minutes. The GP arrived within 11 minutes at the patient's place in 50% of all cases. However, in 50% of all cases the decision making of the GPs before the patient was sent to a hospital required more than 82 minutes. The ambulance arrived within 15 minutes at the patient's place in 90% of all cases. Stabilisation of the patient by the ambulance staff and transport to the hospital took slightly more time. Compared with earlier studies, the patient with a possible myocardial infarction calls for help sooner. Subsequently, in many cases it takes considerable time before the GP refers the patient to a hospital. Further research is needed to improve the diagnostic power of the GP. PMID- 8413731 TI - [Measurement of selective intestinal permeability using a new, simple sugar absorption test]. AB - OBJECTIVE: To study the clinical value of the sugar absorption test (SAT) as a function test of the selective permeability of the small intestine in various intestinal diseases. DESIGN: Inventory of the results of the SAT in a number of patient groups and controls. SETTING: Beatrix Children's Hospital, Groningen and Rijnstate Hospital, Arnhem. METHODS: The SAT was performed in 51 controls (25 children, 26 adults) and in a number of diseases: (suspected) allergy to cow's milk albumin (30 children), (suspected) coeliac disease (86 children and 35 adults), Crohn's disease (25 patients) with ulcerative colitis (9), and pancreatic insufficiency (31) due to cystic fibrosis or chronic pancreatitis. For the SAT, the fasting patient is given a solution of mannitol (M) and lactulose (L) following which the L/M ratio, as an indicator of the selective intestinal permeability, is determined in 5-hour urine by means of gas chromatography. RESULTS: In cow's milk protein allergy, the L/M ratio showed a statistically significant increase in clinically positive cow's milk provocation results, unlike that in clinically negative results. After pretreatment with cromoglycate this difference decreased to non-significant values. The L/M ratio was increased in active coeliac disease (with villous atrophy), first-degree relatives of coeliac disease patients, Crohn's disease, clinically active ulcerative colitis and pancreatic insufficiency. The L/M ratio was not increased in inactive coeliac disease (with normal villi), suspicion of coeliac disease because of short stature, dystrophy and/or aspecific gastrointestinal symptoms with normal villi, and in clinically inactive ulcerative colitis. CONCLUSION: The selective intestinal permeability can be determined by means of the SAT. This could be an important tool for diagnosis and evaluation of therapy in gastrointestinal disorders. PMID- 8413732 TI - [A mixed form of mucocele of the colon]. AB - The term mucocele is applied to an accumulation of mucus within a lumen lined with mucus-secreting epithelium as well as to the structure resulting from leakage of mucus into surrounding connective tissue. The case report is presented of a patient with a bizarre, mixed type of mucocele after perforation of a hemicolon retained 26 years previously, a clinical picture which could not be traced in the literature. PMID- 8413733 TI - [Lyme psychosis]. AB - A woman aged 64 was admitted to the psychiatric department because of a psychotic decompensation with visual hallucinations, disorientation in time and space and associative thinking. On psychotropic drugs the condition failed to improve; subsequently neurological symptoms developed. EEG abnormalities prompted a lumbar puncture. In the CSF a strong plasma cell reaction with atypical cells was observed. The enzyme immunoassay for Borrelia burgdorferi was positive and after treatment with penicillin the psychiatric and neurological signs and symptoms disappeared. From the history which could then be taken it appeared that the patient had been bitten by ticks. Her husband aged 66 years passed through a similar episode of disease. PMID- 8413734 TI - [Liberia; a 'juvenile' fight for control]. PMID- 8413735 TI - [Arthritis as complication of acute meningococcal infection]. PMID- 8413736 TI - [Treatment possibilities in basal cell carcinoma]. PMID- 8413737 TI - [Treatment possibilities in basal cell carcinoma]. PMID- 8413738 TI - [The diagnosis of spondylodiscitis]. PMID- 8413739 TI - [Vaccination against mumps successful]. PMID- 8413740 TI - [Vaccination against mumps successful]. PMID- 8413741 TI - Provider taxes, license fees, and assessments. PMID- 8413742 TI - Why medical students don't choose primary care practice. PMID- 8413743 TI - Current tobacco use policies in Nebraska public schools. AB - A critical effort to reduce the harmful effects of tobacco is to promote tobacco abstinence among the nation's adolescent population. In this regard, the message of tobacco tolerance within the education system of the State of Nebraska was examined. Although the majority of schools surveyed have an encouraging policy regarding student smoking, it has not yet been universally implemented. In addition, the tobacco-use policies for non-students within public school buildings is concerning. The Nebraska Indoor Clean Air Act and the recent Environmental Protection Agency warning on second-hand smoke provides a framework for change. Health care providers can be proactive in encouraging their school districts to promote the goal of a Tobacco-Free Nebraska. PMID- 8413744 TI - Etiology of syncope in 100 patients with associated pale facial appearance. AB - Since publication and discussion of the original patients, I have continued to evaluate patients with loss or near loss of consciousness using the patient's facial color as a guide to underlying etiology. This has consistently provided a quick, reliable method for trying to sort through these difficult cases. Others have similarly noted value of a pale, ashen facial color in diagnosing patients with transient loss of consciousness. I present now my experience with the first 100 patients presenting with syncope and associated pale facial appearance. PMID- 8413745 TI - Decoding the E-code. AB - Injury is a major public health problem. E-coding is an important epidemiologic tool that would allow for quantitative estimates of injury morbidity and mortality. This information could then be used to develop and evaluate injury control interventions. E-coding has been successfully implemented at the county level in Nebraska and efforts should be supported to expand this effort statewide. PMID- 8413746 TI - [Mechanisms of molecular development of human glioma]. AB - Malignant gliomas present a difficult therapeutic problem. Within the last ten years, however, some of the molecular mechanisms have been disclosed that are involved in brain tumour development. According to these results the occurrence of a malignant tumour cell is the result of a multistage process, which involves mutations in several genes. These mutations affect genes regulating cellular proliferation and differentiation, which are called protooncogenes and tumour suppressor genes (anti-oncogenes). Critical alterations in the proteins encoded by these genes may lead to uncontrolled cell proliferation. In astrocytic gliomas mutations of the tumour suppressor gene p53 are common; mutations of the c-erbB gene, encoding the Epidermal Growth Factor receptor (EGF-receptor), frequently affect glioblastomas. The vast majority of glioblastomas show deletions or loss of a copy of chromosome 10. Anaplastic astrocytomas, glioblastomas and oligodendrogliomas frequently carry deletions in one copy of the chromosome 9q and 19q region, respectively. Allelic loss of these regions suggest the presence of a tumour-suppressor gene on these chromosomal loci and a pathogenetic role of anti-oncogene allelic loss in brain tumour development. Molecular studies provide insight into the pathogenesis of brain tumour development. However, they may also have an impact on developing new forms of tumour-specific therapy. The efficacy of such therapeutic strategies is currently being evaluated in clinical studies. PMID- 8413747 TI - [Pregnancy, contraception and epilepsy]. AB - Epilepsy is the most common neurological disease of females in reproductive age. Problems concerning contraception, reproduction, teratogenicity and antiepileptic therapy preceding and during pregnancy are discussed and recommendations made. We underline the advantages of a planned pregnancy with optimal adjustment of antiepileptic drug therapy and recommend prophylactic treatment with folic acid before and during, and with vitamin K towards the end of pregnancy. PMID- 8413748 TI - [Genetic aspects of symptomatic epilepsy based on waking and sleep EEG recordings in siblings]. AB - Waking and sleep EEG-recordings were carried out in siblings of patients with various idiopathic and symptomatic seizure types. Rates of epileptic activity (e.a.) were found in the symptomatic ones varying between 24.1% (Complex partial seizures) and 46.7% (Symptomatic absences). 1/4 to 1/2 of the e.a. was recorded exclusively in sleep, so that sleep recordings are also necessary for such investigations. 2.5-4/sec. spike wave-complexes were predominantly seen; benign foci and photosensitivity were recorded in a smaller number of siblings. More e.a. was observed in idiopathic (72%) than in symptomatic absences (46.7%). On the other hand the same rates (42:41%) as well as almost the same EEG-patterns were found in idiopathic and symptomatic generalized tonic-clonic seizures. When counting the single epileptic discharges more e.a. was seen in siblings of patients with the idiopathic type than the symptomatic one (one discharge every 53 sec.:229.3 sec.). Most e.a. was found in the age group 6-14 years in siblings of all seizure types; therefore, this age dependent penetrance does not depend on the seizure type, but on the recorded spike wave-complexes, benign foci and photosensitivity which occur most frequently in this age range. A multi-factorial mode of inheritance is assumed. PMID- 8413749 TI - [Standardized analysis of visual spatial perception. Construction of the procedure and applications]. AB - Assessment of visuo-spatial perception (VS) in brain-damaged patients. So far, no comprehensive methods for reliable assessment of visuo-spatial deficits after brain damage have been described. The tests usually applied are subtests of intelligence tests, paper-pencil tests or visual-constructive tests which also require motor and planning abilities. Almost all these methods are unsuitable for measuring therapeutic changes and do not reveal specific impairments. The computational method presented here (VS) is suitable for the quantitative assessment of visuo-spatial functions, e.g. subjective visual vertical and horizontal; orientation, length, distance, form and position discrimination as well as line bisection. It can easily be installed on an IBM compatible PC. A clinical standard examination can be performed within 20-30 minutes including numerical and graphical analysis of the results. The method is sensitive for measuring therapeutic changes. Moreover, it offers a wide range of experimental task manipulations for more sophisticated experimental tests. PMID- 8413750 TI - [Local injection treatment with botulinum toxin A in severe arm and leg spasticity]. AB - In patients with predominantly focal spasticity, oral antispastic drugs are relatively ineffective or cause unwanted side effects of central origin. Therefore we treated patients disabled by focal spasticity with local injections of Botulinum-Toxin A (Porton Products BOTOX). Efficacy, dosage, side-effects and injection technique were examined. 11 patients (mean age 48 years) with severe focal spasticity of the flexor muscles of the hand and arm (5 patients), the adductor muscles of the legs (5) or the plantar flexors of the foot (1) due to multiple sclerosis, cervical myelopathy or stroke-related hemi-paresis were treated with BOTOX. Rating scales, including Ashford spasticity scale, pain scale and a hygienic rating scale, were used to evaluate the efficacy. 25 to 30 ng (1000-1200 MU Porton) were injected in the flexor group of the hand or arm and 42 to 50 ng (1680-2000 MU Porton) BOTOX in the adductor group of one leg. 10 of the patients showed an improvement of at least one point on the scales for spasticity, pain and hygiene. Effects could be observed after 4-7 days and lasted for 6-13 weeks. There were no unwanted side-effects. We conclude that BOTOX is an alternative to the systemic application of antispastic drugs. Focal spasticity and pain can be successfully reduced and hygienic care is facilitated. PMID- 8413751 TI - [Neuropathology in the neurosciences. A system in transition]. AB - Neuropathology (Np) is a full member of the neurosciences. As a basic neuroscience it is directed to the behaviour of nervous tissues under pathogenic conditions. The theoretical and methodical core of Np concerns the morphological features of pathological disorders and processes of the nervous system. The goal of Np data presentation is an objective description of the structural changes; their time course as processes, and if possible their causal constellations. Complementary to this analytical task is that of reconstructing the pathological process and at a higher level the conception of pathomorphological entities, e.g. as syndromes. Clinical Np is an alliance of Np with neurology, psychiatry and neurosurgery for representing the structural basis of diseases and the role of morphology in diagnosis and clinical management. Prerequisite for the proper functioning of Np is an integration with these other specialist fields. The clinical neuropathologist therefore has to be in certain respects also a neurologist. The same is true of the alliances of Np with other neurosciences, which is already reflected in recent neuropathological methodology. Detailed training programs are necessary for clinical Np, covering all aspects of its medical and social implications. Enough options should be offered for horizontal flexibility of curricula, futherance of secondary special training and support of good unconventional approaches by junior scientists. PMID- 8413752 TI - [Angiographic occult vascular abnormalities in the area of the pons. Two case reports]. AB - On the basis of two case reports of occult vascular malformation of the pons that had been followed since the first clinical manifestation, clinical picture, diagnosis using modern imaging methods (especially magnetic resonance imaging- MRI) and therapy of this disease are discussed. In addition to a precise localization diagnosis based on typical scans of varying signal intensity, particularly after bleeding, MRI also allows a preoperative qualitative identification of the lesion and provides important information about the dynamics of the process. Further experience with this uncommon clinical category will improve the early diagnosis and selection of an adequate therapy because the technique of stereotactic radiotherapy (gamma-knife) appears to be most suitable. PMID- 8413753 TI - [Necrotizing myopathy with antilipemic agents. Case report and review of the literature]. AB - A 73 year old male who had been prescribed fenofibrate for years developed a slightly asymmetric paraparesis of both lower extremities. CPK values rose to 9800 U/l, EMG of the quadriceps femoris muscle was myopathic. Muscle biopsy revealed a necrotic myopathy. Discontinuation of fenofibrate induced a rapid decline of CPK values, followed by a slower remission of muscular symptoms and persisting pseudo-myotonic discharges in EMG. The spectrum of neuromuscular side effects of cholesterol lowering agents, consisting of myalgia, cramps and reversible CPK elevation, is discussed. Only rarely necrotic myopathies have been described. PMID- 8413754 TI - [Acute cerebellar syndrome in preventive lithium treatment and atypical pneumonia in Q fever]. AB - CNS-involvement in Coxiella burnetii infection is rare. Severe cerebellar symptoms with incomplete restitution were observed in 3 patients who developed Q fever pneumonia associated with a long-term prophylaxis with lithium for manic depressive disorder. Several pathophysiological mechanisms for these complications are discussed and a specific toxic-infectious interaction is suggested. PMID- 8413755 TI - [Cerebellar atrophy and phenytoin poisoning. An MR study]. AB - Phenytoin has been considered a possible cause of cerebellar degeneration, especially after clinical intoxication. Magnetic resonance provides the diagnosis of anatomical structures in the posterior fossa without the limitation of beam hardening artefacts. The aim of this study was to evaluate the relationship of phenytoin medication and cerebellar atrophy in 11 patients with increased serum levels (21.4 micrograms/ml-95.6 micrograms/ml). Five patients had normal cerebellar structures, although three of them had a history of clinical intoxication and all had at least one episode of increased serum level of DPH. The remaining six patients had moderate severe cerebellar atrophy (n = 4) and atrophy of the vermis cerebelli (n = 5). Two of them had never experienced clinical intoxication. There was no correlation between the degree of atrophy and severity of clinical symptoms and evaluation of serum DPH levels (up to four times normal values). There was also no correlation between cerebellar atrophy, duration of epilepsy and frequency of seizures. We conclude that phenytoin overdosage does not necessarily result in cerebellar atrophy and it is unlikely that phenytoin medication was the only cause of cerebellar atrophy in the remaining patients. PMID- 8413756 TI - [Early life experiences and subsequent psychiatric disorders. Contributions of psychiatric epidemiology to a new viewpoint of early childhood]. PMID- 8413757 TI - [Significance of historical and current hallucinogenic research in psychiatry]. AB - Research in experimental psychosis has a long tradition extending through several stages. With the world wide abuse of psychoactive substances it came to an abrupt end. Systematic questions within this framework of research are still unresolved, and research on newly developed compounds is almost completely lacking. A review of the pharmacological and psychological effects of various psychoactive agents is provided, and results of recent experimental studies on arylalkylamines are discussed. It is argued that from a systematic as well as from a methodological viewpoint the traditional approach of experimental psychosis represents a valuable tool in psychiatric research. PMID- 8413758 TI - [The personality of unipolar depressive and bipolar manic-depressive patients]. AB - Starting from central hypotheses on the relationship between personality and affective disorders, the profiles in the Freiburg Personality Inventory (FPI-A) of 23 unipolar depressives, 26 bipolar manic-depressives and 30 healthy controls of the standardization sample were analysed. Assessment by the Freiburg Personality Inventory was carried out at the time of discharge under the instruction to refer the answers to the time before the beginning of the illness. Parallel to the personality assessment the degree of remission was documented by the scales "Depressive Syndrome", "Apathetic Syndrome" and "Manic Syndrome" of the AMDP system and the d2 concentration test. In intraclass covariance analyses the following scales showed significant differences between unipolar depressives, bipolar manic-depressives and normal controls: FPI3 "Depressiveness", FPI8 "Inhibition" and FPIE "Extraversion". The differences cannot be explained by age or sex. In contrast to manic-depressives, unipolar depressives characterize themselves as more depressive, inhibited and introverted. The bipolar manic depressives do not differ from the norms in any scale of the FPI. These findings are compared both with the starting hypotheses and with results reported in the literature. PMID- 8413759 TI - [Is psychoanalytic psychotherapy an "alternative treatment method"?]. AB - Psychoanalytic psychotherapy is a controversial treatment method even though it has an extensive theory and is widely used. Its assumptions continue to be challenged, especially by psychiatrists with a biologic-scientific orientation. Hence it is an appropriate model for examining the transitional area between textbook medicine and what are referred to as "alternative" treatment approaches. The historical development of psychiatry from the end of the 18th century was characterized on the one hand by the increasing influence of biological and technical methods of investigation and on the other by the increasing independence of the citizenry and societal acceptance of subjective emotional experience. The special contribution of Freudian psychoanalytic theory lies in its function as a medicator between these two approaches, whereby it develops analogies between the scientific knowledge of the time and psychic processes. At the same time it provides explanations of emotional illness that the individual can comprehend. This is what is unique about classical psychoanalysis and at the same time makes it problematic, as evidenced, for example, by the fact that so many different schools have developed. The integration of psychoanalysis in the form of psychoanalytic psychotherapy into textbook medicine gives the latter the advantage of being able to reach a clientele that in addition to wanting scientific-causal explanations also has a desire for existential interpretations of psychopathological states. PMID- 8413760 TI - [Modification of cognitive disorders in schizophrenic and schizoaffective psychoses using cognitive therapy in comparison with sociotherapy]. AB - In this study, 24 patients with subacute schizophrenic or schizoaffective psychoses were treated with "Integrated Psychological Therapy for Schizophrenic Patients" (IPT, Roder et al. 1988) and compared with 21 subacute schizophrenic or schizoaffective patients treated with sociotherapy, in respect of cognitive changes. Each therapy was administered at 8 sessions weekly for a period of 6 weeks. Advantages of the cognitively oriented therapy (IPT) appeared significantly greater with regard to interference and short-time memory improvement, and tendentially in respect to overinclusion. Neither type of therapy seems to affect schizophrenic concretism. Cognitive disorders, as reported by the patients themselves, show some remission with both forms of therapy. At 6 and 12 weeks after completion of therapy, differences between the two groups on these measures became insignificant. The findings suggest a quicker reduction of cognitive disorders by IPT in comparison with sociotherapy. PMID- 8413761 TI - [The psychiatric disease of Vincent van Gogh]. AB - From more than 650 letters of van Gogh psychopathologically striking phrases were collected. Their occurrence in the last 18 years of van Gogh's life was observed. The very different interpretations of his symptoms were compiled in a schedule. Finally the case of van Gogh's is used to discuss the borderline between psychosis and epilepsy, a topic which has long been neglected in German psychiatric teaching. PMID- 8413762 TI - [Fatalities in drug-dependent patients. A critical analysis]. AB - The disquieting increase in the mortality of drug addicts observed in recent years cannot be ascribed to only one or a few well defined causes, such as the spread of the HI virus or AIDS, changes in drug consumption habits of multiple dependents or ineffective therapeutic measures. Primarily, an interaction of medical, social and psychological causes is emerging. In a well-defined addict population, 35 deaths of intravenous drug users were registered in the period 1987-1990. By comparison, 7 deaths were recorded in the same geographical area between 1983 and 1986. A report of the UN Commission on Narcotic Drugs shows that in Western Europe, mortality in this high-risk group has doubled annually in the past few years. PMID- 8413763 TI - [Detoxication therapy according to section 64 StGB. Socio-demographic data and therapeutic results of a random 3 year sample]. AB - In the context of a prospective study of alcohol-dependent men, who were committed to compulsory treatment after violations of the law, all patients of a treatment unit in Rheinland/Pfalz during the years 1988-90 were studied. Sociodemographic data, results of psychological testing and records of therapeutic progress of 76 men are evaluated. Besides the evidence of severe social deficits, particular attention is paid to the negative effect of marked delinquency and the additional abuse of drugs and medicaments upon therapeutic outcome. PMID- 8413764 TI - [Bipolar affective disorder and neuropsychiatric complications in a chronic course of thrombotic thrombocytopenic purpura. A case report]. AB - A case report on a 43-year-old female patient with the longest chronic course of TTP known in the literature is presented. The results of comprehensive diagnostic investigations are described. In neuropsychiatric respect there exists a long standing bipolar affective disorder with a course showing a close association to this medical condition. This significant interaction is discussed in relation to the syndromal picture of the phasic bipolar psychosis and the various neuropsychiatric complications associated with recurrent cerebral thrombotic episodes. PMID- 8413765 TI - [Dementia and psychotic symptoms in amyotrophic lateral sclerosis]. AB - We present 2 case histories of patients, treated in psychiatric clinics because of psychotic disorders, in whom further investigation revealed the presence of amyotrophic lateral sclerosis (ALS) and dementia. The causal relationship between psychotic symptoms and dementia on the one hand and ALS on the other hand is discussed. Probably in ALS other functional systems besides motor-neuron pathways are involved. PMID- 8413766 TI - Zinc and cell-mediated immunity in chronic uremia. PMID- 8413767 TI - Regulation of apolipoprotein A-1 and E gene expression in liver and intestine of nephrotic and pair-fed rats. AB - Rats treated with puromycin aminonucleoside (PAN) developed characteristics of the nephrotic syndrome, including albuminuria, hypoalbuminemia and hyperlipidemia. To study the regulation of apolipoprotein (apo) A-1 and apo E gene expression in nephrotic rats, we analyzed the steady-state levels (SSLs) of hepatic and intestinal apo A-1 and apo E mRNA using the Northern technique, and the plasma levels of high-density lipoprotein (HDL) by biochemical methods. Male Wistar rats were treated with PAN and compared with pair-fed and untreated control rats at different stages of disease. Nephrotic rats presented with marked hypoalbuminemia and albuminuria at between 6 and 11 days after PAN treatment. During this stage of disease, plasma levels of HDL were elevated in correlation with an increase of both hepatic and intestinal apo A-1 mRNA. In liver of nephrotic rats, high levels of apo A-1 mRNA together with low levels of apo E mRNA caused an increase in the ratio of apo A-1/apo E mRNA, reaching a maximum 6 days after treatment. Apo E mRNA was barely detected in small intestine of pair fed controls and PAN-treated rats. However, contrary to nephrotic rats, the ratio apo A-1/apo E mRNA was inverted in liver of pair-fed rats due to an increase in apo E mRNA. In conclusion, in nephrotic rats, the SSL of apo A-1 mRNA is increased in liver and small intestine and appears to regulate the plasma levels of apo A-1. These results also suggest a coordinated regulation of the apo A-1 and apo E gene expression in liver of nephrotic and pair-fed rats. PMID- 8413768 TI - Inhibition of glucose utilization in isolated rat soleus muscle by pseudouridine: implications for renal failure. AB - Pseudouridine (psi) is an outstanding nucleoside which is not rebuilt into the tRNA once the parent tRNA is broken down. psi inhibits basal glucose utilization in isolated rat soleus muscle with intact membrane with AD(50)42 mumol/l and Cmax 66%. Psi at concentrations found in renal failure patients inhibits both the insulin- and tolbutamide-stimulated glucose utilization. Its inhibitory activity is partially additive with diltiazem inhibition and nonadditive in the case of magnesium depletion. It is concluded that psi inhibits glucose utilization at the level of Ca modulation in the insulin regulatory cascade. PMID- 8413769 TI - Differential regulation in the expression of hepatic genes in nephrotic and pair fed rats. AB - The messenger ribonucleic acid (mRNA) levels of albumin, fibrinogen, alpha 1-acid glycoprotein (pAGP) and transferrin were analyzed in puromycin aminonucleoside (PAN)-treated (nephrotic) and in pair-fed (PF) rats with the Northern and dot blot hybridization techniques. Albumin mRNA levels in nephrotic and PF rats were 2- and 1.5-fold higher, respectively, than in ad-libitum-fed control (C) rats 6 days after PAN treatment. On day 11, this mRNA in PAN-treated rats was 2.5-fold higher than in PF and 4-fold higher than in C rats. A differential expression at the level of specific mRNAs was also detected for fibrinogen, pAGP, and transferrin in nephrotic and PF rats. On day 6, the fibrinogen and transferrin mRNA levels were significantly higher (p < 0.05) in nephrotic than in PF rats. In contrast, pAGP mRNA levels were normal or low in nephrotic rats and increased 2 fold in PF rats. These studies indicate the differential expression of hepatic genes in nephrotic and PF rats and show that albumin gene expression is only partially regulated by diet during the nephrotic stage of the disease. PMID- 8413770 TI - The effect of dilazep on urinary protein excretion in spontaneous diabetic Chinese hamster. AB - The effect of dilazep, an adenosine potentiator and platelet aggregation inhibitor, on experimental diabetic nephropathy was investigated in spontaneous diabetic Chinese hamster. Prediabetic animals, 8 weeks of age, were divided into two groups. In one group, 5 mg/kg dilazep was injected i.p. once a day. In the other group, saline of the same amount was injected. Age- and sex-matched animals from a nondiabetic subline were used as controls. No difference was observed in body weight, mean blood pressure, fasting plasma glucose and glycated hemoglobin level between diabetic animals with and without dilazep administration throughout the entire period of experiment. Urinary protein excretions in untreated diabetic animals increased significantly compared to those of nondiabetic controls at 8 weeks (17.5 +/- 3.5 vs 2.0 +/- 0.1 mg/day), and at 24 weeks (25.3 +/- 5.1 vs 2.7 +/- 0.1 mg/day) of experiment. In diabetic animals with dilazep treatment, urinary protein excretions (4.1 +/- 0.7 at 8 weeks and 13.1 +/- 2.9 mg/day at 24 weeks of experiment) were significantly suppressed compared to those in untreated diabetic animals. Significant thickening of glomerular basement membrane (GBM) was observed in diabetic animals both with and without dilazep administration at 24 weeks of experiment compared to that in nondiabetic controls. The number of anionic sites in GBM, stained by polyethyleneimine, was reduced in untreated diabetic animals, but was not different in dilazep treated animals compared to that in nondiabetic controls. It was concluded that dilazep administration suppressed urinary protein excretion in diabetic Chinese hamster possibly through the preservation of charge barrier of the glomerulus. PMID- 8413771 TI - Unusual findings in a myeloma kidney: a light- and electron-microscopic study. AB - A case of IgD myeloma together with the existence of myelofibrosis is presented. More interesting is the concurrent presence of cast nephropathy, light chain deposition and amyloidosis in the kidney. Peculiar light-microscopic, immunohistochemical, immunofluorescence and ultrastructural findings were also noted. The possible mechanisms and implications for such findings were discussed. This case was analyzed together with a review of local pictures of renal involvement in multiple myeloma. PMID- 8413772 TI - An adult case of Fanconi syndrome due to a mixture of Chinese crude drugs. AB - We examined a 35-year-old male case of acquired Fanconi syndrome induced by a mixture of Chinese crude drugs. Renal glycosuria, hypokalemia, hypophosphatemia, metabolic acidosis, a low threshold of tubular bicarbonate excretion, and generalized aminoaciduria were observed after the patient had taken the drugs for 6 months. When he stopped taking them, all laboratory data improved. He took the drugs again on his own judgment, leading to a second bout of Fanconi syndrome. This is the first case in which Chinese crude drugs have been known to cause acquired Fanconi syndrome. PMID- 8413773 TI - Serum aluminum transport and aluminum uptake in chronic renal failure: role of iron and aluminum metabolism. AB - Several factors have been blamed for increasing gastrointestinal absorption of aluminum. The likely role of iron metabolism was suggested some years ago. As iron and aluminum share many chemical properties, it is reasonable to think they also share biological pathways. The aim of this study was: (a) to evaluate serum aluminum transport and its relationship with iron-binding capacity, and (b) to investigate aluminum hydroxide absorption as a function of iron and aluminum. We investigated 127 patients with chronic renal failure undergoing hemodialysis in a study divided into two phases: phase 1, a basal study to investigate serum iron and aluminum status, and phase 2 in which an aluminum absorption test was performed. In phase 1, we found that the lower basal serum iron and iron transferrin saturation the greater serum aluminum (p < 0.001). In phase 2, we found a negative relationship between serum aluminum increments after the test and basal levels of serum aluminum and iron (r = -0.70; p < 0.001). These results suggest that the amount of either aluminum or iron carried by transferrin may influence the transferrin capacity to bind the other element and also may modulate, together with other factors, the gastrointestinal absorption of iron and aluminum. PMID- 8413774 TI - Efficacy of dual lumen jugular venous catheter hemodialysis when venous lumen is used as arterial lumen. AB - In 12 patients who underwent hemodialysis by a dual lumen jugular venous catheter, urea recirculation rate was measured during conventional treatment under normal conditions (R1), as well as when the venous lumen of the catheter was used as arterial lumen and the arterial as venous lumen (R2). Although the mean value of R2 was, as expected, higher than R1 (8.25 +/- 2.7 versus 4.25 +/- 1.00; p = 0.0004), it was still within the acceptable rate. We conclude that during hemodialysis by a dual lumen jugular venous catheter, efficient treatment can be provided when the venous is used as arterial lumen. PMID- 8413775 TI - Pharmacokinetics and dialysability of warfarin in end-stage renal disease. PMID- 8413776 TI - Urinary endothelin and sodium excretion in essential hypertension. PMID- 8413777 TI - Piperacillin-induced acute interstitial nephritis. PMID- 8413778 TI - Vascular injury in acute renal failure due to leptospirosis is not associated with antineutrophil cytoplasmic antibody. PMID- 8413779 TI - Laparoscopic rescue of dysfunctional Tenckhoff catheters in continuous ambulatory peritoneal dialysis patients. PMID- 8413780 TI - Direct tubular toxicity of hymenoptera venom. PMID- 8413781 TI - The apparent "epidemic" increase in the incidence of renal failure from diabetic nephropathy. PMID- 8413782 TI - Disturbed intracellular calcium homeostasis as a major reason of vascular contractile dysfunction in hypertension of renal origin. PMID- 8413783 TI - Effect of pleurodesis with autoblood on hydrothorax due to continuous ambulatory peritoneal dialysis-induced diaphragmatic communication. PMID- 8413784 TI - Acute renal failure due to bilateral ureteral hematomas complicating anticoagulant therapy. PMID- 8413785 TI - Vasculitis associated with non-Hodgkin's lymphoma. PMID- 8413786 TI - Death risk in CAPD patients. The predictive value of the initial clinical and laboratory variables. AB - The characteristics, survival rate and risk factors associated with death in patients with end-stage renal failure treated with chronic ambulatory peritoneal dialysis (CAPD) were studied. This is a retrospective study of a cohort of 206 patients, from which the follow-up was complete in 190 patients (92%). Only 16 patients (8%) were lost. The study group is composed of 118 males and 88 females, with a mean age of 39 +/- 15 years. The origin of the renal disease was: unknown in 90 patients (44%); diabetes mellitus in 50 (24%); systemic lupus erythematosus in 16 (8%); obstructive uropathy in 15 (7%); glomerulonephritis in 14 (7%), and miscellaneous in 21 (10%). The average follow-up was 12 +/- 11 months. At the end of study, 66 patients were dead (32%). CAPD was discontinued in 12 (6%). Thirty eight patients (18%) received kidney transplantation. The survival rate for the whole group was 67 and 48% at 1 and 3 years, respectively. Multivariate survival analysis according to the Cox proportional-hazard model showed that the most powerful predictor associated with high risk of death was low serum albumin levels. According to the Cox model other independent variables significantly associated with increase in the probability of death while on CAPD were advancing age, low serum creatinine concentrations and elevated serum cholesterol levels. These results indicate that the risk factors associated with death in CAPD patients are similar to those observed for hemodialysis patients and suggest that using simple laboratory measurements at the enrollment in CAPD the relative risk of death for each patient can be estimated. PMID- 8413787 TI - The influence of salt sensitivity on the blood pressure response to exogenous kallikrein in essential hypertensive patients. AB - In order to verify the influence of salt sensitivity on the blood pressure response to orally administered kallikrein, we evaluated the efficacy of glandular kallikrein (derived from porcine pancreas) in 28 essential hypertensives (21 males and 9 females) aged between 40 and 62 years. After a placebo run-in period, the patients were assigned to receive oral kallikrein therapy (150 IU 3 times a day; n = 18 patients) or placebo (n = 10 patients) over a period of 8 days in a random double-blind fashion. In the salt-resistant patients (n = 8), kallikrein administration did not modify blood pressure levels. In the same group, natriuresis increased significantly after the treatment [from 94.51 +/- 10.76 to 111.65 +/- 23.19 mEq/24 h (mmol/24 h), p < 0.039]. In the salt sensitive patients (n = 10), blood pressure decreased with the kallikrein therapy (systolic: from 158.50 +/- 9.20 to 144.50 +/- 10.12 mm Hg, p < 0.005; diastolic: from 99.50 +/- 2.16 to 90.0 +/- 3.67 mm Hg, p < 0.024). In the same patients, urinary Na+ excretion increased considerably after the kallikrein treatment (from 101.07 +/- 18.36 to 134.34 +/- 18.27 mEq/24 h, p < 0.0001). Therefore, our data indicate that the oral kallikrein administration reduces blood pressure levels only in the salt-sensitive hypertensives. In both the salt-sensitive and the salt resistant groups a marked increase in the 24-hour urinary excretion of sodium was observed after the kallikrein treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413788 TI - Influence of calcium intake on calcitriol levels in idiopathic hypercalciuria in children. AB - Twenty-four children with idiopathic absorptive hypercalciuria (IAH) and a control group (CG) of 11 healthy children were studied. Plasma 1,25 hydroxyvitamin D3 (calcitriol), parathyroid hormone (PTH), calcium (Ca) and phosphate (P) levels were measured during dietary manipulation. The three diets analyzed were: (A) calcium-restrictive diet (400 mg/1.73 m2/day) for 7 days; (B) supplemented diet (1,000 mg/1.73 m2/day) for 3 days; (C) supplemented diet continued for 15 days. The IAH group had higher levels of serum calcitriol than the control group for all three diets. Serum calcitriol levels in the IAH group decreased in diet B compared to diet A, and returned to levels observed with diet A during diet C. Serum Ca, P and plasma PTH levels did not vary throughout the study in either group. In IAH, two subgroups were observed. In one, serum calcitriol levels were elevated and in the other, serum calcitriol levels were not different from the controls. This second group had a lower P and maximum rate of tubular reabsorption of phosphate per 100 ml of glomerular filtrate than the IAH group with elevated serum calcitriol levels and the control group. These results suggest that IAH in children may be related both to increased serum calcitriol levels and to an altered Tmp/GFR. PMID- 8413789 TI - Serum lipoprotein (a) levels in maintenance hemodialysis patients. AB - To further understand lipoprotein (a) [Lp(a)] and atherosclerosis, we measured serum Lp(a), lipoprotein, and apolipoprotein levels in 55 patients (males, 24-73 years old) on maintenance hemodialysis, and compared them with those of 82 controls (males, 21-81 years old). The serum Lp(a) levels in patients on maintenance hemodialysis were significantly higher than those of the normal controls, while serum total cholesterol (TC), high-density lipoprotein cholesterol, (HDL-C), HDL2-C, HDL3-C, apolipoprotein (apo) Al, apo All levels, and lecithin-cholesterol acyltransferase (LCAT) activities were significantly (p < 0.05) reduced in the patient group. The frequency distribution of serum Lp(a) levels in the patients was different from that in the control group, and no prognostic tendency of serum Lp(a) levels was noted by the etiology of renal failure as histologically determined by the renal biopsies. In the patient group, we also found that serum Lp(a) levels negatively correlated with serum triglycerides (TG) and total protein (TP) concentrations (p < 0.05), but no correlation was found between the duration of hemodialysis therapy or patient age and the serum levels of TC, TG, apo B and Lp(a) levels when tested for simple regression. Significant (p < 0.05) positive correlations were also found between TP and serum TG, apo B, and LCAT activities. These opposing tendencies of Lp(a) and serum TG, apo B, when measured against TP concentrations, indicate that serum TP levels may not affect serum lipoprotein and Lp(a) levels in the same direction. These data suggest that hemodialysis or end-stage renal disease itself, rather than hypoproteinemia, may hold the key to high serum Lp(a) levels in hemodialysis patients. PMID- 8413790 TI - Variability of glomerular response to injury. PMID- 8413791 TI - Iodine-induced hypothyroidism in patients on regular dialysis treatment. AB - Hypothyroidism with a serum concentration of thyroid-stimulating hormone (TSH) above 40 microU/ml was noted in 3 (3.2%) of 93 patients on regular hemodialysis or continuous ambulatory peritoneal dialysis. These 3 patients had no history of thyroid disease and were receiving no medication known to influence thyroid function. They had habitually eaten iodine-rich foods and showed an enlarged thyroid gland with a preserved radioactive iodine uptake and a markedly elevated serum inorganic iodine (II) level. In all 3 patients, both thyroidal microsomal antibody and thyroglobulin antibody titers measured by hemagglutination methods were less than 100, and TSH-binding inhibitory immunoglobulin was negative. Moreover, histologically no lymphocytic infiltrations were observed. With only iodine restriction, serum TSH level markedly decreased from 44.6 to 3.6 microU/ml in case 1, from 90.6 to 3.2 microU/ml in case 2 and from 43.2 to 9.4 microU/ml in case 3 in parallel with decreases in the serum II level. These results suggest that at least in an area like Japan, where the daily intake of iodine is high, iodine-induced hypothyroidism may be induced in patients undergoing regular dialysis treatment even in the absence of apparent underlying thyroid disease. PMID- 8413792 TI - Pattern of interleukins in minimal-change nephrotic syndrome of childhood. AB - Assays of interleukin-1 (IL-1) and IL-2 were done in supernatants from phytohaemagglutinin-activated lymphocyte cultures from 10 children suffering from minimal-change nephrotic syndrome (MCNS) to assess their role in the aetiopathogenesis of this disorder. Increased levels of IL-1 and IL-2 had been found in supernatants from patients having MCNS compared with controls, suggesting a significant role of these cytokines in the immunopathogenesis of proteinuria in this syndrome. PMID- 8413793 TI - Toxicity of free radicals to mesothelial cells and peritoneal membrane. AB - We studied the toxicity of free radicals to human mesothelial cells in vitro and to the peritoneal membrane of rats during peritoneal dialysis. Free radicals cause damage to mesothelial cells as measured by release of cytosolic markers such as 86Rb and lactate dehydrogenase. Vitamin E neutralized the toxic effect of free radicals in vitro. Human mesothelial cells exposed over 6 h to a mixture of essential and nonessential amino acids in medium are more vulnerable to the cytotoxic effect of free radicals than control cells exposed to medium alone. Cells exposed previously to glucose or glycerol are less vulnerable than controls. In rats free radicals generated intraperitoneally by a xanthine xanthine oxidase system induce changes in peritoneal permeability similar to those observed during peritonitis: loss of ultrafiltration, increased glucose absorption from the dialysate and augmented transperitoneal loss of albumin. In addition lipids in the peritoneum became peroxidated. The addition of vitamin E to the peritoneal fluid with xanthine-xanthine oxidase prevents peroxidation of lipids and the subsequent loss of ultrafiltration. Our results show that free radicals may exert a potentially toxic effect on the peritoneal membrane during peritonitis. In such circumstances the addition of free radical scavenger to the dialysis fluid may preserve intact structure and function of peritoneum. PMID- 8413794 TI - Atrial natriuretic peptide and hemodynamic changes after concentrated ascitic fluid reinfusion in cirrhotic patients. AB - In order to assess the role, if any, of atrial natriuretic peptide (ANP) and hemodynamic changes in diuretic response to concentrated ascitic fluid reinfusion (CAFR) in cirrhotic patients, we studied 10 patients with liver cirrhosis before and after aspiration, concentration and intravenous reinfusion of their ascitic fluid. Basal ANP levels were higher than in normal subjects, but not related to the degree of water and sodium excretion. CAFR induced a variable degree of ANP changes and diuretic response, and these two parameters were strictly correlated to each other. No major changes of cardiac output and systemic vascular resistances were observed after CAFR. In conclusion, the apparent resistance to ANP diuretic action observed in basal condition in cirrhotic patients seems to revert after expanding blood volume by CAFR. PMID- 8413795 TI - Effect of lactate infusion on renal transport of purine bases and oxypurinol. AB - To investigate whether or not lactic acid inhibits the renal transport of oxypurines and oxypurinol, we administered physiological saline containing 0.2 mol sodium lactate to 5 normal subjects intravenously. Lactate infusion decreased the fractional clearance of uric acid, but the fractional clearances of hypoxanthine, xanthine and oxypurinol were not affected. These results suggest that uric acid and lactic acid share the renal transport system of organic acids but hypoxanthine, xanthine and oxypurinol do not. It is further suggested that allopurinol treatment is reasonable in subjects with hyperuricemia accompanied by hyperlactatemia since only the urinary excretion of uric acid and not oxypurines (hypoxanthine and xanthine) was inhibited by lactate infusion. PMID- 8413796 TI - Artificial simulation of renal stone formation. Influence of some urinary components. AB - The effect of natural admixtures occurring in human urine (citrate, pyrophosphate and glycosaminoglycans) on the precipitation of stone-forming compounds was studied. Experiments were carried out under conditions closely simulating the early stages of renal stone formation. Among the studied admixtures, citrate was determined as the most effective substance preventing the phosphate particle formation. Indeed, in the presence of citrate, some calcium oxalate monohydrate crystals were found. Pyrophosphate induced the formation of calcium oxalate dihydrate crystals. Phosphate crystals appeared at pH 6 and never at pH 5. The easy formation of phosphate particles supports the hypothesis that these crystals represent a very important heterogeneous nucleus-initiating oxalocalcic calculus formation in the kidney. Reported results also indicated uric acid as a significant heterogeneous nucleus of calcium oxalate monohydrate crystals at urinary pH equal or lower than 5 and the important role of bacteria in increasing the organic detritus deposited on the solid surfaces. PMID- 8413797 TI - Suppressive effect of an oral sorbent on the accumulation of p-cresol in the serum of experimental uremic rats. AB - Serum p-cresol and phenol are markedly accumulated in uremic patients. To determine if an oral sorbent (AST-120) can decrease their serum concentrations in the uremic state, an oral sorbent was administered to experimental nephrectomized uremic rats. In uremic rats fed with oral sorbent, the serum concentration and the urinary excretion of p-cresol were markedly and significantly lower than those in control uremic rats, while those of phenol tended to be low in the uremic rats with oral sorbent as compared with control uremic rats. The concentrations of serum creatinine and blood urea nitrogen were significantly decreased in the uremic rats with oral sorbent. These findings demonstrate that oral sorbent adsorbs especially p-cresol in the intestine and prevents the accumulation of p-cresol in the serum of uremic rats. PMID- 8413798 TI - Development of renal failure in endotoxemic rats: can it be explained by early changes in renal energy metabolism? AB - Endotoxin shock not only causes renal failure, endotoxemia also leads to metabolic impairment, resulting in energy shortage and loss of cellular integrity; therefore, we tested the hypothesis that early changes in renal metabolism contribute to the development of acute renal failure during endotoxin shock. Endotoxin (Escherichia coli 127B8; 8 mg/kg from t = 0 to 60 min) was infused in three groups of 8 rats, in which renal biopsies were taken at t = 30, 50 and 90 min, respectively; a fourth group (n = 8) served as control. In the biopsies, glucose, lactate, ATP, ADP, AMP and creatine phosphate concentrations were determined. Renal plasma flow (RPF) and glomerular filtration rate (GFR) were measured from the clearances of 131I-hippurate and 125I-thalamate, respectively. We also assayed urine flow (V; catheter in the bladder), cardiac output (CO), blood pressure (MAP), heart rate (HR) and arterial lactate, glucose and creatinine concentrations. During the first 30 min of endotoxemia, we found no systemic hemodynamic or biochemical changes. From t = 30 to t = 90, CO and MAP decreased to 59 and 70%, respectively, while HR and serum levels rose to 110 and 800%, respectively (p < 0.05), indicating progression of shock. Renal function clearly deteriorated from t = 30; at t = 90 RPF, GFR and V had decreased by 86, 84 and 86%, respectively, plasma creatinine being 193% of the baseline value (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413799 TI - [Skull base surgery using intraluminal shunt tube]. PMID- 8413800 TI - [Risk factors for severe subarachnoid hemorrhage following aneurysmal rupture]. AB - Major reduction in disability and death from severe subarachnoid hemorrhage (SAH) can probably be brought about more effectively by prevention than by better medical or surgical treatment. Identification of the major risk factors for severe SAH should facilitate the preventive efforts. As risk factors for severe SAH, a patient's age, sex, location and size of aneurysms, hypertension, rebleeding including minor leak, and cigarette smoking were selected. The relation between the incidence of severe SAH, a surgical results, and the risk factors was examined in 81 severe SAH cases. Hypertension, rebleeding from aneurysms, large aneurysm exceeding 1.0 cm/sec in size were closely connected to severe SAH. Treatment of hypertension, acute surgery for aneurysms and aggressive surgery for unruptured large aneurysms were the only hope for achieving substantial reduction in the incidence of severe SAH and for improving the surgical results. PMID- 8413801 TI - [Newly developed blocking balloon catheter for PTA of internal carotid artery]. AB - We have developed a new blocking balloon system for percutaneous transluminal angioplasty (PTA) of the internal carotid artery (ICA). A latex balloon (BALT) is attached on the top of a Superselector infusion-type catheter (TORAY) which is used as a blocking balloon catheter. It can be navigated into the distal part of the ICA under torque control as well as flow control by withdrawing the core wire. Our PTA procedure is as follows, 1) insert the sheath catheter 8-9 F in size, 2) insert the PTA balloon catheter (Accent balloon, Cook) with a blocking balloon catheter through the sheath catheter, 3) navigate the blocking balloon into the distal part of the ICA, 4) introduce the PTA balloon to the stenotic portion after occlusion of the distal ICA by the blocking balloon, 5) PTA, 6) wash the lumen of the ICA with saline, 7) deflate the blocking balloon and withdraw the system. We performed PTA for 5 ICA lesions in 4 cases and got successful dilatations for all of them without complications. Our blocking balloon system is useful for performing the PTA of the ICA safely. PMID- 8413802 TI - [Neurological deterioration induced by giant intracranial aneurysms even though these aneurysms had not ruptured; mural hemorrhage and intraluminal thrombosis]. AB - The authors report two cases of intracranial giant aneurysms inducing acute neurological deterioration even though rupture of the aneurysms had not occurred. Neurological aggravation was attributable to the acute swelling of the aneurysmal mass after intramural hemorrhage in one case, and the formation of rapid and massive intraluminal thrombosis in another case. MRI confirmed these mechanisms in both cases. MRI is very useful for demonstrating mural and intraluminal pathologies involving giant aneurysms. PMID- 8413803 TI - [Clinical study in patients with perimesencephalic subarachnoid hemorrhage of unknown etiology]. AB - We have recognized a characteristic distribution of cisternal blood in 10 (43%) of 23 patients with subarachnoid hemorrhage (SAH) of unknown etiology. On the initial CT examination undertaken within 48 hours of the onset, blood from 10 patients was found to be more densely distributed in the cisterns around the brainstem. In this study, clinical characteristics and CT findings in those 10 cases were carefully evaluated and the CT findings were compared to those of 416 patients with aneurysmal SAH (anterior circulation aneurysm 368 cases, posterior circulation aneurysm 48 cases). There were seven men and three women, with an age ranging from 39 to 64 years (average age, 50.6 years). The follow-up period ranged from 4 to 45 months (average follow-up period, 23 months). Neurological grade (WFNS) on admission was I in 9 cases and III in one case. None of the patients suffered symptomatic vasospasm, hydrocephalus or rebleeding. All the patients had favourable outcome and were categorized as good recovery according to the Glasgow Outcome Scale. In comparison with the cases of aneurysmal SAH, especially in comparison with 48 cases with SAH caused by posterior circulation aneurysm, 43 cases could be easily distinguished on CT. The other 5 cases showed almost the same pattern of SAH on CT, but 4 cases could be differentiated by either the extension of SAH to the interhemispheric fissure or the presence of intraventricular hemorrhage. Only one case could not be differentiated on CT. Thus SAH located only around the brainstem differs from aneurysmal SAH in its clinical course, and in distribution and severity of bleeding on CT. This could be recognized as a new clinical entity and could be called benign SAH. PMID- 8413804 TI - [Evaluation of the postoperative regrowth of the acoustic neurinomas]. AB - Although recent advances in microsurgical removal of acoustic neurinomas has made it possible to preserve, morphologically, the cochlear nerve as well as the facial nerve, functional impairment of the facial nerve is often encountered following total removal of the tumor. In order to avoid such functional morbidity, a surgical procedure can be proposed in which the tumoral capsule attached to a functional structure such as the facial nerve is intentionally left. Along with this surgical procedure and postoperative course observation, postoperative regrowth of the tumor should be precisely evaluated. A few recent reports about this problem have been published since CT scan and MRI have become available. The authors reviewed 66 cases in which postoperative evaluation by high resolution CT scan of the acoustic neurinomas has been carried out in their department more than one year after surgery. Multiple factors such as age at onset, sex, preoperative period, tumor size, vascularity, extent of the tumor removal, residual site, and pathological findings on HE stain were studied in relation to the tumor regrowth. Four cases showed regrowth of the tumor, and one of these tumors was reoperated on. The percentage of regrowth was 7.5% among the tumors which were not totally removed. No significant risk factors for tumor regrowth were noticed in these cases, but tumor regrowth tended to be found in younger patients. The residual tumors which showed regrowth were always located in the internal auditory meatus. In this study, it is apparent that, percentagewise, tumor regrowth is not so prevalent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413805 TI - [Neuropsychological outcome of head injury in children]. AB - Wechsler Intelligence Scale for Children-Revised (WISC-R) and Yatabe-Guilford personality test were administered to 31 children who had been hospitalized for head injury and made a GR or MD by the Glasgow Outcome Scale (GOS). The type of lesion, as defined by CT scan categories, was an important factor to prognosticate the outcome of intellectual function. The IQ, especially performance IQ, of acute subdural hematoma (EDH) or severe diffuse brain injury (DBI) was lower than that caused by other types of lesion. Several children demonstrated improvement in IQ level during the initial year. The difference between the IQ of the children who could return to previous school life and that of the children who could not was significant. One of the causes of difficulty in returning to previous school life is decreasing IQ and personality change such as social disadaptability. Neuropsychological evaluation is important in predicting school recovery and deciding proper neuropsychological rehabilitation. PMID- 8413806 TI - [Falx metastasis of thymic carcinoma: a case report and review of literature]. AB - A patient, a 72-year-old male, with a thymic carcinoma spreading to the extrathoracic region, is reported. he had undergone a thoracotomy and received radiation-and chemotherapy. Four months later, the patient was noticed to have a mild left sided hemiparesis. Further evaluations revealed metastasis of thymic carcinoma to the bone and intracranial region. Surgical treatment for intracranial lesion was performed, and the patient's left sided hemiparesis was cleared. Thymic carcinoma is uncommon, and its intracranial metastasis is rare. We found 27 previous reports of intracranial metastasis of thymic carcinoma and malignant thymoma. This time, we report a case of thymic carcinoma with intracranial (falx) metastasis, and discuss the concept of thymic carcinoma and thymomas in a review of the literature. PMID- 8413807 TI - [A case of true human tail accompanied with spinal lipoma]. AB - Human tails have been attributed to a disturbance in fetal tail regression which normally occurs at the gestation age of about 6 weeks. To date, more than 100 cases of human tails have been reported. However, reports of true human tails, which involve the coccygeal vertebrae, are rare. We recently encountered a patient with a true human tail which involved the coccygeal vertebrae and was accompanied by lumbar spinal lipoma and spina bifida. A four-year-old boy was brought to our clinic with complaints primarily of painful mass. The boy had no neurological abnormalities. Physically, a tail bone projected, slightly in the lumbar area, with a linear depression in the center. A hard tail bone was palpable subcutaneously. A soft mass was palpable in the lumbar region, which was accompanied by hemangioma on the superficial layer of the skin in this region. On X-ray films, the tail bone lacked the normal curvature and it projected linearly in the posterior direction. CT scans revealed spina bifida at the level below L2. MRI disclosed spina bifida (at the level below L2), spinal lipoma and a tethered cord. During surgery, the tail bone was first resected over a distance of one and a half vertebral bodies. The lumbar tumor, which continued into the spinal canal, was removed as completely as possible after incision of the dura mater. To free the tethered cord, the tensioned, hypertrophic filum terminale was dissected. Although the morphological diagnosis of this condition is easy, the high incidence of complication by other anomalies makes it essential to perform thorough preoperative examinations with CT and MRI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413808 TI - [A rare case of middle fossa neuroepithelial cyst accompanied by chronic subdural hematoma]. AB - We reported a rare case of middle fossa neuroepithelial cyst accompanied by chronic subdural hematoma. An eight-year-old boy was admitted to our hospital due to headache, disturbed consciousness and right hemiparesis. CT disclosed a left marginal high density in the left subdural space and remarkable midline shift. MRI showed a hematoma in the left to temporal tip extending to the chronic subdural hematoma. Radiological examinations revealed a cystic lesion with chronic subdural hematoma. An operation was performed to remove the hematoma. Intraoperatively, the hematoma in the cyst and the chronic subdural hematoma were observed. Histopathological diagnosis of the cyst wall was neuroepithelial cyst. The relationship between the neuroepithelial cyst and chronic subdural hematoma was discussed. PMID- 8413809 TI - [Subarachnoid hemorrhage and pontine hemorrhage followed by an embolization procedure of left occipital giant arteriovenous malformation: a case report]. AB - A 42-year-old woman suddenly developed headache and nausea on July 26, 1991, and the computed tomography (CT) scan showed a moderate-sized hematoma in the left occipital lobe. After one month's conservative treatment, she had recovered to a neurologically intact state. Cerebral angiography demonstrated a giant arteriovenous malformation fed by enlarged branches of the left posterior cerebral artery as well as small branches arising from the middle cerebral artery, anterior cerebral artery and the meningeal branches of the middle meningeal artery and the occipital artery. Preoperative embolization was planned on February 24, 1992. During an attempt at catheterization of the basilar artery and the left posterior cerebral artery with a balloon catheter and a Tracker-18 catheter, the patient complained of an intensification of her headache, nausea and vomiting. So the embolization procedure was stopped. The CT scan taken immediately at that time showed a severe subarachnoid hemorrhage (SAH). She became comatose about 40 minutes later. CT scan taken next day revealed also a complication of the pontine hemorrhage. Neurologically, she had gradually recovered and could communicate with some simple words 3 months after SAH. The total removal of the AVM was performed on May 26, 1992. Postoperative course was uneventful. She showed rapid and remarkable improvement in her neurological state suggesting that the blood flow in the surrounding brain area had been corrected. A blood deficit had no doubt been caused when blood had been stolen by the giant AVM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413810 TI - [Convexity angiomatous meningiomas in a mother and a daughter without evidence of neurofibromatosis]. AB - The convexity angiomatous meningiomas that occurred in a mother and a daughter without any evidence of neurofibromatosis (NF) were reported. The 73-year-old mother was admitted to our clinic because of an episode of generalized convulsion and a five-month history of gait disturbance. She had the signs/symptoms of intracranial hypertension and frontal lobe dysfunctions. Computed tomography (CT) revealed a left frontal enhanced mass with a small intratumoral cyst and a remarkable perifocal edema. Angiography showed tumor stain fed from the external carotid artery. Frontal craniotomy was performed and a dark red tumor was totally resected. The nodular-surfaced tumor had adhered loosely to the dura mater. The coarse vascular meshwork and an intratumoral cyst were observed on the cut surface. When the patient was discharged she was able to leave the hospital on foot, but she died of acute pancreatitis in the local hospital. Histological examination of the tumor showed rich vasculatures with focally whorl-formed cells. Most tumor cells had intracytoplasmic microcysts. The pathological diagnosis, WHO's classification, 1991, was angiomatous meningioma. The patient's 41-year-old daughter was admitted due to an episode of fainting. All laboratory data were within normal limits, including the normal karyotype of the peripheral blood leukocytes. Papilloedemata were the only signs of neurological deficit. A CT scan and magnetic resonance images showed a left frontal convexity mass and angiography displayed the tumor strains from the middle meningeal artery. The convexity meningioma similar to her mother's was totally removed. The histological diagnosis was angiomatous meningioma, again.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413811 TI - [A case of glial microtumor treated with brachytherapy using MRI guided stereotactic system]. AB - A 34 year old male suffered from convulsion on his right side with loss of consciousness. Neither plain nor enhanced CT could clearly demonstrate a lesion. However, a Gd-DTPA enhanced T1 weighted image (T1WI) demonstrated an enhancing lesion (2 cm in diameter) in the left frontal lobe including the motor cortex. This lesion was not identified on plain T1WI. A T2 weighted image (T2WI) showed a high intensity lesion without mass effect slightly larger than the enhanced area on T1WI. This lesion was suspected to be a glial microtumor, but, because it was difficult to differentiate it from a cerebral infarction, it was confirmed to be a neoplasm by means of an MRI guided stereotactic biopsy. The histological diagnosis of the frozen section was a grade 2 astrocytoma. Subsequently brachytherapy with 192Ir seeds was performed, and about 80Gy of interstitial irradiation was given to the edge of the enhanced area on MRI. The minimum dose in the high intensity area on T2WI was about 50Gy. The patient was discharged with no neurological deficits. The Gd-DTPA enhanced lesion decreased in size six months after the treatment. MRI guided stereotactic system makes it possible to target a high intensity lesion on T2WI in brachytherapy. Brachytherapy using this system is considered to be useful in the treatment of glial microtumor in an area in which it is difficult to identify it on CT. PMID- 8413812 TI - [Venous thrombosis after closed head injury: a report of two cases presenting as intracranial hypertension]. AB - Although sinus thrombosis occurs owing to various causes, sinus thrombosis after closed head injury is rarely reported. The authors encountered 2 cases with sinus thrombosis in whom intracranial hypertension occurred after closed head injury. In case 1, left abducens and oculomotor palsy developed as a false localizing sign, together with symptoms of intracranial hypertension 10 days after head injury. Using angiography, we made a diagnosis of stenosis of the superior sagittal sinus. Since the symptoms were aggravated under conservative treatment, V-P shunt was performed, but impaired vision was not improved. In case 2, symptoms of intracranial hypertension occurred 10 days after head injury. We performed magnetic resonance angiography (MRA) and made a diagnosis of stenosis of the bilateral transverse sinuses. The symptoms improved under conservative treatment. The use of conventional angiography has been the most reliable for a diagnosis of sinus thrombosis, but MRA will become useful from now on. For diagnosis, it is important to suspect sinus thrombosis after closed head injury. Surgical decompression is required for patients in whom symptoms are aggravated under conservative treatment. PMID- 8413813 TI - [Spinal extradural angiolipoma: a case report]. AB - Growth of angiolipoma of the central nervous system is a rare occurrence. Only about 40 cases can be collected in the literature. A 67-year-old man was admitted because of intermittent severe pain in both legs after walking. He had chillness in both legs during the past 5 years which gradually worsened. MR image showed a spinal mass lesion at the level of Th12 to L1. The lesion was isointense on T1 weighted image and slightly hyperintense on T2-weighted image. The tumor was located on the left side of the spinal cord and was markedly enhanced after intravenous Gd-DTPA. At the operation, a soft, dark-red mass was found situated in the epidural space at the level of Th12 to L1, which was totally removed. On histological examination, the tumor was shown to be mainly composed of mature fatty cells and of numerous blood vessels with enlarged lumens. Postoperative course was uneventful and the symptoms subsided during the following few months. PMID- 8413814 TI - Paraventricular and supraoptic nuclei of the hypothalamus are not equally important for oxytocin release during stress. AB - The relative importance of the paraventricular (PVN) and the supraoptic nuclei (SON) for the secretion of oxytocin was evaluated by comparison of stress-induced oxytocin release under normal conditions, in the absence of vasopressin and/or corticoliberin (CRF). We introduced an incomplete anterolateral cut (iALC) around the mediobasal hypothalamus designed to leave intact the SON-neurohypophysial connections but to inflict damage to the nerve fibers from the PVN. The studies were performed in conscious cannulated rats using immobilization as the stress stimulus. Stress-induced oxytocin release was found in heterozygous Brattleboro rats as well as in homozygous animals lacking vasopressin, yet in the latter it was less pronounced and in both cases it was prevented by iALC. In Wistar rats, stress-induced oxytocin release was markedly reduced after iALC and absent after PVN lesion. Both hypothalamic interventions failed to influence basal oxytocin levels and resulted in a similar reduction of ACTH release. It is concluded that a functional diversity exists between the hypothalamic magnocellular nuclei. At least in relation to immobilization stress, the PVN is essential for stress induced oxytocin release and it is evident that the SON without the PVN cannot preserve oxytocin secretion during stress. PMID- 8413815 TI - Ontogenesis of the sexual dimorphism of growth hormone secretion by perifused rat hemipituitaries. AB - The aims of the present study were to investigate the sexual dimorphism and the role of sex steroids in GH secretion at the pituitary level, and to evaluate the ontogenesis of these effects. Towards these aims we used an in vitro perifusion system of hemipituitaries under a simulated milieu of hypothalamic factors: two 3 min pulses of GHRH at 3-hour intervals were separated by continuous flow of somatostatin. Rat GH was measured in 2.4-min fractions and analyzed by the pulse analysis program PULSAR. Pulses were similar in prepubertal male and female rats, but sexual dimorphism was evident in adults. In adult males, who had undergone neonatal gonadectomy, GH pulse amplitude and area under the curve (AUC) were lower compared to control. When gonadectomy had been performed at a prepubertal age, the pulse amplitude was still lower, but the AUC was not different from control. The gap between orchiectomy at neonatal and prepubertal age indicates the perinatal imprint, which induces an increase in AUC. Neonatal testosterone treatment of intact female rats had no effect on GH secretion by adult pituitaries. In neonatally gonadectomized female rats, under neonatal testosterone treatment, the pulse amplitude increased. A similar increase was observed after neonatal gonadectomy without testosterone treatment. We conclude that the sexual dimorphism of GH secretion is partially induced at the pituitary level and its response to the hypothalamic hormones. We assume that a neonatal imprint effect of testosterone in the male induces primarily an increase in AUC in response to GHRH. The imprint in females influences the GH pulse amplitude and AUC. PMID- 8413816 TI - Role of a T-type calcium current in supporting a depolarizing potential, damped oscillations, and phasic firing in vasopressinergic guinea pig supraoptic neurons. AB - Guinea pig magnocellular neurosecretory cells (MNCs) of the supraoptic nucleus (SON) were studied using the in vitro slice preparation. Intracellular recordings were made with biocytin-filled electrodes, permitting immunocytochemical identification of the recorded cells as arginine vasopressin- (AVP) versus oxytocin- (OT) containing. Only AVP cells displaying a depolarizing potential (DP) fired phasically. The DP was associated with a transient inward current measured in voltage clamp, which exhibited a number of properties of the T-type calcium current: activation threshold of -64 mV, time course of up to 250 ms, blockade by nickel and augmentation by barium chloride. This current has not been reported previously in SON neurons. The T-type current (IT) was always associated with a damped oscillation of the membrane following the offset from hyperpolarizing steps. In all cells tested, an apamin-sensitive afterhyperpolarization (AHP) was observed, similar to the calcium-dependent potassium current (IK, Ca) described in rat SON and other CNS regions. Therefore, as with other CNS regions displaying damped oscillations, guinea pig SON cells possess both an IT and an IK, Ca. We have previously described an Ih activating at hyperpolarized potentials in these cells, which depolarizes the membrane to a range in which the IT and IK, Ca can interactively support oscillations. In summary, the IT and associated depolarizing potential appears to be a requisite feature for phasic firing in AVP cells of guinea pig SON. PMID- 8413817 TI - Modulation of the hypothalamo-pituitary-adrenocortical responses to cytokines in the rat by lipocortin 1 and glucocorticoids: a role for lipocortin 1 in the feedback inhibition of CRF-41 release? AB - Our recent studies indicate that lipocortin 1 (LC1), a putative second messenger protein for the anti-inflammatory steroids in peripheral tissues, may also contribute to the regulatory actions of the glucocorticoids on the hypothalamo pituitary-adrenal axis. In the present study we have used in vitro and in vivo models to compare the effects of adrenalectomy, LC1 and dexamethasone on the cytokine-induced secretion of the 41-amino acid corticotrophin releasing factor (CRF-41) and arginine vasopressin (AVP) by the rat hypothalamus. In addition, western blot analysis was used to examine the influence of dexamethasone on the expression of LC1 in the hypothalamus. In vitro, interleukins- (IL-) 1 alpha (100 and 200 pg/ml), 1 beta (0.5 and 1.0 ng/ml) and 8 (0.25-1.0 ng/ml) readily initiated the release of CRF-41 and AVP from hypothalami removed from intact rats. IL-6 (10 and 20 ng/ml) was also an effective CRF-41 secretagogue but, unlike the other interleukins tested, it was ineffective with regard to AVP. Adrenalectomy 7-14 days prior to autopsy increased significantly (p < 0.01) the magnitude of the CRF-41 responses to IL-1 alpha, IL-1 beta and IL-6 but not to IL 8. In contrast however, while hypothalamic tissue from adrenalectomized rats, unlike that from intact animals, responded to IL-6 (5-20 ng/ml) with a pronounced hypersecretion of AVP, the AVP responses to IL-1 alpha and IL-1 beta were largely unaffected by adrenalectomy as too were those to IL-8. The marked increases in CRF-41 and AVP release from hypothalami from adrenalectomized rats initiated in vitro by IL1 alpha, IL-1 beta, IL-6 and IL-8 were readily overcome by preincubation of the tissue with dexamethasone (10(-7) M). In addition, the steroid caused 'externalization' of two species of immunoreactive (ir-) LC1 (37 and 58 kDa) by the hypothalamic cells but failed to influence the total LC1 content of the tissue. The inhibitory effects of dexamethasone on the cytokine induced release of CRF-41 in vitro were mimicked by LC1 (10 ng/ml) which alone had no effect on the basal release of the peptide. However, unlike dexamethasone, LC1 failed to influence the concomitant release of AVP from the hypothalamic tissue elicited by IL1 alpha, IL-1 beta or IL-8 and potentiated that evoked by IL 6.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8413818 TI - Rapid neurophysiological effects of corticosterone on medullary neurons: relationship to stress-induced suppression of courtship clasping in an amphibian. AB - Courtship clasping of females by male roughskin newts (Taricha granulosa) is rapidly blocked by exposure to corticosterone (CORT). This behavioral effect appears to result from CORT binding to a receptor in neuronal membranes. The present study investigated effects of intraperitoneal CORT administration on neurophysiological properties of extracellularly recorded single medullary neurons in acutely prepared newts. CORT produced multiple neurophysiological effects that emerged within 3 min of injection and increased in magnitude during the next 20-30 min. Spontaneously active and sensory-responsive neurons showed a decline or cessation of firing concomitant with a loss of sensory responsiveness, especially to cloacal pressure, a clasp-facilitating stimulus in behaving newts. After CORT administration, reticulospinal neurons that were backfired (antidromically activated) by spinal cord stimulation, exhibited reduced antidromic action potential amplitude, slowed rates of spike generation and other indications of reduced excitability. Comparable effects of CORT were also evident in newts with a premedullary brainstem transection, indicating a direct hormone action on the caudal neuraxis. Dexamethasone (DEX), a glucocorticoid that binds poorly to the CORT membrane receptor and has little effect on clasping, had little or no direct neurophysiological effect, but DEX injection 30 min before CORT interfered with the neurophysiological action of CORT. The rapidity, time course and specificity to CORT of these neurophysiological effects are consistent with mediation through the CORT membrane receptor. In addition, the pattern and dose sensitivity of these neurophysiological actions plus their occurrence in the medulla, suggest that they could underlie the CORT effect on courtship clasping. PMID- 8413819 TI - Secretion of polypeptide growth factors by human nonfunctioning pituitary adenoma cells in culture. AB - The growth-promoting activities of tumor-conditioned media (TU-CM) obtained from 23 cultured human nonfunctioning pituitary adenomas were studied in vitro. TU-CM obtained from adenoma cell cultures increased both cell counts (range: 108-179%; control = growth in serum-free medium = 100%) and 3H-thymidine incorporation (112 139%) of rat pituitary cell cultures, indicating that TU-CM contains growth stimulating substances. TU-CM was also able to stimulate the growth of normal fibroblasts (3H-thymidine incorporation: 164-178%; cell counts: 145-157%) and endothelial cells (3H-thymidine incorporation: 131-149%; cell counts: 181-217%), suggesting the presence of - possibly angiogenic-growth factors that act on these cell types. However, the growth of hormone-producing cells was also stimulated, since TU-CM increased 3H-thymidine incorporation into rat pituitary cells in the presence of D-Val-MEM, a medium specifically inhibiting growth of fibroblasts. Addition of neutralizing antibodies against transforming growth factor alpha (TGF alpha), epidermal growth factor (EGF), insulin-like growth factor I (IGF-I) and basic fibroblast growth factor (bFGF), either alone or in different combinations, reduced the growth-promoting activity of TU-CM on rat pituitary cells (range: 96 71%; control = growth effect of TU-CM without antibodies = 100%), strongly indicating the presence of these growth factors in TU-CM. All 4 antibodies together completely inhibited the growth-stimulatory activity of TU-CM, strongly suggesting that these growth factors play the major role among growth-stimulating substances in TU-CM. This is the first study giving evidence that TGF-alpha, EGF, IGF-I and bFGF are secreted by nonfunctioning adenoma cells indicating that the growth factors could be involved in growth regulation of pituitary adenomas by paracrine or autocrine mechanisms. PMID- 8413820 TI - Lack of glucocorticoids sustains the stress-induced release of noradrenaline in the anterior hypothalamus. AB - The release of endogenous noradrenaline in the anterior hypothalamus was studied with microdialysis perfusion in freely moving rats that were subjected to immobilization stress. Experiments were carried out in sham-adrenalectomized and adrenalectomized rats that were first given drinking water containing corticosterone for 5 days following surgery and then switched to a corticosterone free diet the day before stress application. One group of these adrenalectomized animals was injected with dexamethasone. Basal release of noradrenaline collected in 20-min fractions was similar in the three groups of animals and averaged 24 fmol. The recovery of the probe was about 10%. In sham-adrenalectomized rats application of 20-min immobilization stress increased noradrenaline release to 310% of baseline in the sample collected during stress application. A significant increase (+ 175% of baseline) was still observed in the next 20-min sample. Subsequent values were all identical to baseline. In adrenalectomized rats lacking exogenous corticosterone the stress-induced release of noradrenaline was prolonged with noradrenaline levels remaining elevated for 2 h after the onset of stress. The total noradrenaline release during this entire period was about 2.5 times higher in adrenalectomized than in sham-operated rats. However, the maximal increase during the period of immobilization was not significantly affected. Treatment with dexamethasone prevented the prolonged increase in noradrenaline release but did not affect the increase during the period of stress. While glucocorticoids do not appear to affect the increased release of NA in the anterior hypothalamus during the period of stress, they act to limit the duration of this activation after the application of stress.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413821 TI - Glucocorticoid regulation of angiotensinogen gene expression in discrete areas of the male rat brain. An in situ hybridization study. AB - The regulation of angiotensinogen gene expression by glucocorticoids has been described in several studies. Kalinyak and Perlman reported on a 60% increase of angiotensinogen expression in the rat brain after dexamethasone treatment with a single, high-dose injection. The purpose of the present study was to investigate whether a general upregulation of angiotensinogen expression or a region-specific upregulation underlies these findings. By using in situ hybridization and computer-assisted microdensitometry we evaluated the regional changes in angiotensinogen expression following dexamethasone treatment. Angiotensinogen expression was strongly stimulated in several areas (medial septum and locus ceruleus), while only moderately in others (hypothalamus, medulla). An interesting finding is the difference in glucocorticoid receptor regulation among the circumventricular organs. The subfornical organ displayed no changes in angiotensinogen-mRNA, whereas the area postrema showed an increase. Furthermore, the angiotensinogen expression in the median eminence decreased substantially. The nature of these effects will form the subject of further investigations. In conclusion we demonstrated an overall increase but certain heterogeneities in angiotensinogen expression after dexamethasone treatment, a pattern which suggests different degrees of glucocorticoid receptor regulation of the angiotensinogen gene in certain areas of the brain, probably mainly dependent on the degree of glial glucocorticoid receptor presence. PMID- 8413822 TI - Ethanol alters N-methyl-DL-aspartic acid-induced secretion of luteinizing hormone releasing hormone and the onset of puberty in the female rat. AB - In the present study, we have evaluated the effects of ethanol (ETOH) on luteinizing hormone releasing hormone (LHRH) secretion induced by N-methyl-DL aspartic acid (NMA) in vitro and the ability of NMA to induce precocious puberty in vivo. For the in vitro experiments, the basal and NMA-stimulated release of LHRH from arcuate nucleus-median eminence (AN-ME) fragments obtained from immature female rats was measured by RIA following static incubation in medium consisting of Krebs-Ringer bicarbonate glucose buffer. Control vials contained medium only and the test vials contained medium plus ETOH doses of 30, 50, and 70 mM. These data demonstrate that ETOH did not alter basal LHRH release, but dose dependently blocked (p < 0.01) the NMA-induced release of the peptide during anestrus, as well as first proestrus and estrus. For the in vivo experiment, 26 day-old females began receiving saline or saline-ETOH (3 g/kg) solution by gastric gavage daily at 12.30 p.m. Each day at 2.00 and 4.00 p.m., each of the animals received subcutaneous injections of a 40-mg/kg solution of NMA in saline. Other control animals received saline gastrically, as well as subcutaneously. The timing of puberty was assessed in all animals by monitoring vaginal opening (VO) and first diestrus (D1). The mean (+/- SEM) age at VO for animals in the non-ETOH group, which received NMA, was 31.7 +/- 0.40 days. Vaginal smears revealed that D1 occurred at a mean (+/- SEM) age of 33.0 +/- 0.42 days.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413823 TI - Differential effect of short- and long-term lithium treatment on m-CPP-induced neuroendocrine changes in rats. AB - Intravenous administration of the serotonergic agent m-chlorophenylpiperazine (m CPP) to rats produced increases in plasma prolactin and corticosterone and a decrease in plasma growth hormone concentrations. Short-term (2-4 days) and long term (21-23 days) lithium treatment did not affect baseline levels of prolactin, corticosterone or growth hormone. Long-term lithium treatment accentuated m-CPP's effect on plasma prolactin and corticosterone but not on growth hormone levels. On the other hand, short-term lithium treatment attenuated m-CPP's effect on plasma corticosterone but not on plasma prolactin or growth hormone levels. These findings demonstrate a differential effect of the duration of lithium treatment on m-CPP-induced neuroendocrine changes. One possible explanation for this differential effect may be that different 5-HT receptor subtypes are involved in mediating different neuroendocrine functions and, furthermore, these different 5 HT receptor subtypes may be affected differentially following lithium treatment. Alternatively, lithium-induced changes in other neurotransmitter systems may be responsible for this differential effect. PMID- 8413824 TI - Opiate receptor subtype involvement in the stimulation of prolactin release by beta-endorphin in female rats. AB - The prolactin secretory response to beta-endorphin and the involvement of opiate receptor subtypes in this response was determined in both diestrous and postpartum, lactating female rats. The involvement of the mu-, delta- and/or kappa-site was determined by administering specific antagonists for each of these sites prior to beta-endorphin. beta-Funaltrexamine (beta-FNA, 1 or 5 micrograms) was administered to block mu-sites, ICI 154,129 (5, 10 or 25 micrograms) blocked delta-sites and nor-binaltorphimine (norBNI, 8 micrograms) blocked kappa-sites. The ability of beta-FNA and ICI 154,129 to block prolactin secretion following morphine administration was also determined. A dose response study for beta endorphin indicated that beta-endorphin, at doses as low as 25 ng, was a potent stimulus for prolactin release producing an increase in prolactin that mimicked the suckling-induced prolactin increase. In addition, all three antagonists were capable of antagonizing the stimulatory effect of beta-endorphin in both diestrous and postpartum female rats. These results indicate that beta-endorphin is a potent stimulus for prolactin secretion and that these three opiate receptor subtypes interact to produce its stimulatory effect on prolactin release. PMID- 8413825 TI - Met-enkephalin secretion from mixed cultures of hypothalamic neurons and astrocytes. AB - Release of the endogenous opioid pentapeptide, met-enkephalin, from primary cultures of dissociated fetal rat hypothalamic cells was studied using an assay system which could both measure and differentiate between free met-enkephalin and the larger enkephalin-containing peptides (ECPs), which are the processing intermediates of the enkephalin precursor. The cultures were maintained in fully defined, serum-free medium and contained both neurons and astrocytes. Free met enkephalin was secreted from the cultures in significant quantities in response to nonspecific depolarisation with 56 mM potassium, by a mechanism dependent upon extracellular calcium. Under basal conditions, barely detectable amounts of free peptide were released, whereas ECPs were secreted in significant quantities which were not reduced by the removal of extracellular calcium. As the period of culture increased, so did the quantitative importance of this constitutive ECP secretion, relative to the stimulated release of free peptide. Treatment of the cultures with the cytotoxic agent, cytosine arabinoside, attenuated this temporal increase of ECP secretion, whilst leaving the stimulated release of free met enkephalin relatively unaffected. This suggested that the met-enkephalin secretion seen within the cultures reflected the presence of at least two distinct enkephalinergic cell types and that the change in the nature of the secreted enkephalin was at least in part, due to the proliferation of one of these cell populations. These results are consistent with secretion of met enkephalin from both neurons and astrocytes within these cultures. We propose that the neurons secreted essentially fully processed peptide in a regulated manner, whilst the mitotic glial cells constitutively secreted non- or partially processed precursor peptides.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413826 TI - Bacterial lipopolysaccharide induction of IL-6 in rat telencephalic cells is mediated in part by IL-1. AB - Interleukin-6 (IL-6) appears in the cerebrospinal fluid (CSF) of patients with acute infection of the central nervous system, and in the brains and CSF of experimental animals following systemic or intracerebral injection of bacterial endotoxin (Escherichia coli lipopolysaccharide, LPS). Since LPS is known to induce secretion of interleukin-1 (IL-1) in many cell types including those of the brain, and IL-1 can induce IL-6 in brain tissue it appeared reasonable to postulate that the effects of LPS on IL-6 production were mediated through IL-1 induction. To test this hypothesis, the effects of IL-1 receptor antagonist (IL 1Ra) on LPS and IL-1-induced IL-6 secretion were tested in a mixed brain cell culture from 17-day fetal rat, after 12-14 days in culture. IL-6 secretion was induced by IL-1 beta in a concentration as low as 1 x 10(-10) M (p = 0.0008); addition of IL-1Ra was shown to inhibit IL-1-induced changes by 87% (p = 0.0012) at a molar ratio of 100:1, and by 100% at a molar ratio of 1,000:1, LPS stimulated IL-6 secretion progressively over the concentration of 1-100 ng/ml (p = 0.0001). LPS 10 ng/ml-induced IL-6 secretion was inhibited by 66% by IL-1Ra in a concentration of 1,000 ng/ml (p = 0.0077). The inhibitory effect of IL-1Ra was not significantly greater even when used at a concentration of 5,000 ng/ml.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413827 TI - Influence of male rats on the luteinizing hormone-releasing hormone neuronal system in female rats: role of the vomeronasal organ. AB - Olfactory information processed by the vomeronasal system is reported to influence reproductive functions in a variety of mammals. The present studies were designed to determine if male-associated cues affect the luteinizing hormone releasing hormone (LHRH) neuronal system, and, if so, to determine the extent to which these cues are processed by the vomeronasal organ (VNO). Ovariectomized rats underwent VNO removal (VNX) or sham surgery (VN-Sham). Forty-eight hours after estrogen priming (5 micrograms), they were subjected to one of the following treatments: repeated mating, repeated exposure to male-soiled bedding or repeated exposure to clean bedding. In animals treated for 180 min, coronal brain sections were double labelled for Fos protein and LHRH. An intense Fos immunoreactivity was induced following mating in the majority of LHRH neurons in the VN-Sham females, whereas removal of the VNO significantly suppressed the mating-induced Fos staining. Exposure of female rats to male-soiled bedding or clean bedding did not induce appreciable Fos immunoreactivity in LHRH neurons. Following 90 min of mating or exposure to bedding, blood samples were assayed for luteinizing hormone (LH). Mating stimulated the release of LH in VN-Sham females, while the removal of the VNO significantly suppressed the mating-induced LH release. Exposure of the females to male-soiled bedding or clean bedding did not induce an LH surge. The present results demonstrate that male-originating sensory cues (i.e. repeated mating) can influence the LHRH neuronal system, as evidenced by the presence of Fos immunoreactivity in LHRH cell bodies, and indicate that this effect is mediated through the VNO to a certain extent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413828 TI - Reduction in the arginine vasopressin responses to metoclopramide and insulin induced hypoglycemia in normal weight bulimic women. AB - A low plasma arginine vasopressin (AVP) responsiveness to hypertonic saline infusion has been described in bulimic women. At present, it is unknown whether this phenomenon is peculiar for the osmotic regulation of AVP secretion or whether it represents an aspect of a more general disorder of AVP secretion in bulimia nervosa. In order to answer these questions, in the present study the AVP responses to metoclopramide (MCP) (20 mg in an i.v. bolus) and insulin (0.15 IU/kg)-induced hypoglycemia were tested in normal weight bulimic women and in weight- and age-matched normal women. Basal AVP concentrations were similar in normal and bulimic women. In the normal controls, plasma AVP levels rose 2.6 times after MCP and 2.2 times in response to hypoglycemia. Both AVP increments were significantly lower in bulimic patients. In this group, plasma AVP levels rose 2 times after MCP and 1.8 times in response to hypoglycemia. When data of the MCP and insulin tolerance test were combined, regression analyses showed a significant positive correlation between AVP peak responses to MCP and hypoglycemia in the bulimic group. These data show an impaired AVP response to different releasing stimuli in bulimia, suggesting that a more general disorder than a simple change in the sensitivity to osmotic stimulation affects the AVP secretory system in bulimic patients. It is likely that bulimic subjects are affected by a neuroendocrine alteration in the control of AVP secretion, whose mechanisms are still unknown. PMID- 8413829 TI - Estrogen induces synaptic plasticity in adult primate neurons. AB - The number of axosomatic synapses, the length of the synaptic plates and the perimeter of the post synaptic neuronal perikarya were assessed on thin sections from the infundibular hypothalamic nucleus and from the ventrobasal thalamus of 3 adult ovariectomized African green monkeys that were treated with estradiol valerate and of 3 control animals that were injected with vehicle. Estradiol valerate treatment resulted in a 61% [corrected] decrease in the number of axosomatic synapses in the infundibular hypothalamic nucleus. The length of the synaptic plates and the perimeter of the postsynaptic cells were not affected by the hormonal treatment. The decrease in the number of axosomatic synaptic inputs in the infundibular hypothalamic nucleus was accompanied by a significant increase in the glial ensheathing of neuronal somas. No effect of the hormonal treatment was detected in the ventrobasal thalamus. The results indicate that estrogen may induce glial and synaptic plasticity in the hypothalamus of adult primates. PMID- 8413830 TI - Progesterone, but not LHRH or prostaglandin E2, induces sequential inhibition of lordosis to various lordogenic agents. AB - In experiment I we studied the capacity of progesterone (P) and two nonsteroidal agents that activate lordosis, but do not bind to the progestin receptor (PR), i.e. luteinizing hormone-releasing hormone (LHRH) and prostaglandin E2 (PGE2) to induce sequential inhibition (SI) in ovariectomized estradiol-primed rats. The administration of 1 mg P, 5 micrograms LHRH or 100 micrograms PGE2 induced significant lordosis within 4 h. An injection of 1 mg P, 24 h after the administration of the above lordogenic agents, induced significant lordosis in rats pretreated with LHRH or PGE2, but not in those pretreated with P. Thus, only P induced SI (p < 0.025). In experiment II we investigated if progestin-induced SI results in a reduced capacity of the subjects to respond only to P or to other lordogenic agents. The synthetic progestin norgestrel (400 micrograms administered 24 h earlier) significantly reduced the responsiveness to P (p < 0.01), LHRH (p < 0.01), PGE2 (p < 0.025) and dibutyryl cyclic AMP (db cAMP p < 0.01). Results suggest that SI is triggered only by agents that bind to the PR (experiment I) and that it decreases the responsiveness of rats not only to P but also to other lordogenic agents (experiment II). PMID- 8413831 TI - Serotonin, catecholamines and metabolites in discrete brain areas in relation to lordotic responding on proestrus. AB - Levels of serotonin (5-HT) and metabolite, 5-hydroxyindole-acetic acid (5-HIAA), dopamine and metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), and norepinephrine (NE) were measured in microdissected brain areas at 14.30 and 19.30 h on diestrus and proestrus in rats. Lordosis, sexual behavior, was exhibited only by proestrous females at 19.30 h. The content of monoamines and/or metabolites changed from afternoon to early evening of diestrus in a number of brain regions. On proestrus, during the time when females became sexually receptive, additional changes appeared and many diestrous changes were reversed or amplified. Proestrus-specific changes were found in areas providing the descending circuit for regulation of lordosis, the medial preoptic nucleus (mPOA), ventromedial nucleus of the hypothalamus (VMN) and midbrain central gray (MCG), and in areas outside of endocrine control centers such as parietal cortex (Ctx) and dorsal raphe nucleus (DR). Thus, these data are consistent with previous studies showing functional changes in monoaminergic transmitters during the day/night cycle and during hormonal induction of neuroendocrine events. NE levels increased between 14.30 and 19.30 h on proestrus in most brain areas, ranging from 20% increases in mPOA and Ctx to a 220% increase in the DR. NE levels decreased on proestrus in the VMN. The direction of proestrous changes in NE were reversed in some, but not all of the areas, between the afternoon and evening of diestrus. Dopamine was detectable in all areas sampled, and the metabolites HVA and DOPAC were detectable in some but not all areas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413832 TI - Evidence that nitric oxide can act centrally to stimulate vasopressin release. AB - Nitric oxide (NO) is the endothelium-derived relaxing factor, which causes relaxation of vascular smooth muscle. NO synthetase, the enzyme for the synthesis of NO from its precursor L-arginine, is also widely distributed in neurons in the brain, and it has been suggested that NO may serve as an important neuromodulator. Because NO synthetase is present in the hypothalamus in relatively high concentration, we have determined whether NO can affect the release of vasopressin in conscious, chronically prepared rats. The intracerebroventricular (i.c.v.) injection of S-nitroso-N-acetylpenicillamine (12.5 and 25 micrograms; SNAP), that spontaneously breaks down to form NO, caused transient dose-related increases in the plasma vasopressin concentration of 1 and 2 microU/ml (p < 0.01), respectively. In control experiments in which N acetylpenicillamine (25 micrograms), the precursor for the preparation of SNAP, was injected i.c.v. there was a small, 0.4 microU/ml, increase (p < 0.01) in the plasma vasopressin level. The i.c.v. injection of L-arginine (0.5 and 1 mg), also the precursor for the biosynthesis of NO, resulted in dose-dependent increases in the plasma vasopressin concentration similar in magnitude to those caused by SNAP. When D-arginine (1 mg), which cannot serve as a substrate for NO synthetase, was injected i.c.v., there was only a slight delayed increase in the plasma vasopressin concentration. Thus, NO can act centrally to stimulate vasopressin release and may serve as a neuromodulator in the control of vasopressin release. PMID- 8413833 TI - Amino acid levels in the hypothalamus and response to N-methyl-D-aspartate and/or dizocilpine administration during sexual maturation in female rats. AB - Amino acid concentration in the anterior preoptic area and medial basal hypothalamus was determined by HPLC in female rats: (1) at 16 (prepubertal) vs. 30 (peripubertal) days of age and (2) after N-methyl-D-aspartate (NMDA) or dizocilpine (MK-801) administration in both groups. 30-day-old rats had higher levels of aspartate (Asp; 24%), glutamate (Glu; 49%) and glycine (Gly; 44%) and lower levels of taurine (Tau; 43%) than 16-day-old rats. In 16-day-old rats, NMDA (30 mg/kg, s.c., 10 min) increased the Glut concentration (48%). This effect was prevented by MK-801 pretreatment (1 mg/kg, s.c., 1 h), which did not modify amino acid concentrations per se. In 30-day-old rats, NMDA treatment increased Glut (24%) and asp (42%) levels. MK-801 pretreatment abolished NMDA-induced changes and reduced Tau (26%) and Gly (30%) levels. MK-801 administration alone reduced the concentration of Glut (39%), Asp (54%), Tau (33%) and Gly (31%). It is concluded that both (1) the concentration of Asp, Glu, Gly and Tau and (2) the changes induced by NMDA receptor activation or blockade are different at 16 vs. 30 days of age. The existence of a tonic (positive) control on amino acid levels linked to the NMDA receptor which would be immature or absent at 16 days of age is suggested. PMID- 8413834 TI - Steroid hormones and receptors of the GABAA supramolecular complex. I. Benzodiazepine receptor level changes in some extrahypothalamic brain areas of the female rat following sex steroid treatment. AB - The effects of sex steroid hormones on the different receptor binding sites of the GABAA molecule remain unclear. In this report we have demonstrated, using autoradiography techniques, that the distribution pattern of the benzodiazepine receptors (a component of the GABAA molecule) in some extrahypothalamic brain regions is altered by both in vivo and in vitro sex steroid hormone treatment. In vivo administration of the sex steroids estradiol and progesterone induced a significant change in [3H]flunitrazepam (benzodiazepine agonist) binding levels in the amygdala, and cortico and posterior brain nuclei of the female rat. In fact, elevated and diminished receptor-binding levels were obtained in the corticomedial amygdala nucleus and in the pontine central gray matter respectively, following the administration of estradiol. Significant hormonal effects were also shown for animals that received only a progesterone dose, as demonstrated by the increased and decreased receptor levels in the basolateral amygdala nucleus and cortex lamina VI and in the substantia nigra pars reticulata, respectively. It was interesting, at this point, to investigate whether the hormone effects on [3H]flunitrazepam binding changes might be mediated through a GABA-dependent activity, because the benzodiazepine and GABAA receptors are coupled to a chloride ion channel in an allosteric manner. When 50 microM GABA was added to the incubation medium, substantially altered binding levels were recorded in animals that received progesterone replacement therapy only. The GABA-induced progesterone effects both increased substantially the binding levels in the oriens-pyramidalis CA1 layer of the hippocampus and in the intermediate gray layer of the superior colliculus as well as reducing receptor levels in the substantia nigra pars reticulata.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413835 TI - Steroid hormones and receptors of the GABAA supramolecular complex. II. Progesterone and estrogen inhibitory effects on the chloride ion channel receptor in different forebrain areas of the female rat. AB - The inhibitory effect of female sex steroid hormones on the binding of [35S]t butylbicyclophosphorothionate [35S]TBPS to the chloride ion channel receptor in different forebrain areas of the female rat proved to be of a differential nature. The in vivo administration of estradiol and estradiol+progesterone were responsible for substantially lower chloride ion channel receptor levels in brain areas that contain elevated steroid receptors, such as the medial preoptic area, the cortico-medial amygdala nucleus, the vertical limb diagonal band-medial septal nucleus and the cortex lamina V. The administration of progesterone alone reduced receptor levels in the oriens-pyramidalis CA1 layer of the hippocampus, caudate putamen, cortex lamina VI (brain areas that contain little if any steroid receptors) and in the lateral and basolateral amygdala nucleus (brain sites that contain noninducible progesterone receptors). On the basis of the progesterone inhibitory activity on the chloride ion channel receptors, it was important to investigate whether progesterone per se or whether the potent progesterone metabolites 3 alpha-hydroxy-5 alpha-dihydroprogesterone (3 alpha, 5 alpha-THP) and 3 beta-hydroxy-5 beta-dihydroprogesterone (3 beta,5 beta-THP) were involved in the binding level changes, and to establish the specific brain sites where these steroid effects occur. In fact, in vitro addition of the 5 alpha-reduced progesterone metabolite produced even greater depressive effects on [35S]TBPS binding not only in the same brain areas as the in vivo progesterone replacement therapy but also in some sites that provided significant receptor level changes following the sequential administration of estradiol+progesterone. However, when the 5 beta-reduced metabolite was tested on the binding of [35S]TBPS to the chloride ion channel receptor, only the basolateral amygdala nucleus, the cortex lamina VI and the dorsolateral septal nucleus exhibited changes. Because the steroid-mediated chloride ion flux is regulated in a GABA-dependent manner, we next checked for the type of GABA effects on the chloride ion channel receptor levels and found that GABA not only intensified the 3 alpha,5 alpha-THP inhibitory effects but, together with this progesterone metabolite, was also involved in binding changes in the vertical limb diagonal band-medial septal nucleus. It is interesting to note that the GABA effects on 5 beta-metabolite induced receptor changes were not of the enhancing type, but tended, rather, to be inhibitory.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8413836 TI - BRL 46470 potently antagonizes neural responses activated by 5-HT3 receptors. AB - The effect of a novel 5-HT3 receptor antagonist, BRL 46470, has been studied on two electrophysiological models for 5-HT3 receptors: grease-gap recordings from rat isolated vagus nerve and whole-cell patch-clamp recordings from mouse neuroblastoma-rat glioma NG108-15 cells. Its action on the rat vagus nerve was compared to that of four other 5-HT3 receptor antagonists. On the rat vagus, BRL 46470 reduced the maximum depolarizing response to 5-HT in a concentration dependent manner with an IC50 of 0.3-1.0 nM, but the EC50 for 5-HT was not appreciably affected. This action was similar to that of granisetron and ICS 205 930, but differed from that of GR38032F and (+)-tubocurarine which produced clear rightward shifts of the concentration-response curve to 5-HT. The 5-HT-induced fast inward current of voltage-clamped NG108-15 cells was also antagonized by 1 nM BRL 46470 in an insurmountable manner. In contrast to (+)-tubocurarine, the action of BRL 46470 on the rat vagus nerve and NG108-15 cells did not readily reverse on washing with antagonist-free medium. It is concluded that BRL 46470 is a potent, insurmountable 5-HT3 receptor antagonist on the rat vagus and NG108-15 cells. PMID- 8413837 TI - [3H]paroxetine binding in rat frontal cortex strongly correlates with [3H]5-HT uptake: effect of administration of various antidepressant treatments. AB - Paroxetine is a selective and potent inhibitor of 5-hydroxytryptamine (5-HT) uptake into serotonergic neurones. [3H]Paroxetine binding to rat frontal cortex was of high affinity with a high percentage of specific binding. The binding data of both competition and saturation studies fitted a single site binding model. [3H]Paroxetine binding was potently inhibited by the selective 5-HT uptake inhibitors. In addition, a very good correlation was demonstrated between the ability of twenty-three compounds to inhibit [3H]paroxetine binding to rat frontal cortical membranes and [3H]5-HT uptake into rat frontal cortical synaptosomes. These data support the view that [3H]paroxetine binds to a single site which corresponds to the 5-HT uptake site. Using this ligand, the effects of repeated administration of antidepressant drugs with a wide range of pharmacological actions and electroconvulsive shock on 5-HT reuptake sites were examined. [3H]Paroxetine binding parameters (Kd and Bmax) were unaltered by all treatments. It would, therefore, appear that antidepressant therapy does not produce adaptive changes in 5-HT uptake sites. PMID- 8413838 TI - Cholinesterase inhibitor effects on extracellular acetylcholine in rat cortex. AB - A microdialysis technique was used to sample acetylcholine (ACh) from the cerebral cortex of conscious rats. We thus investigated the effects of systemically administered cholinesterase inhibitors (ChEI) such as physostigmine (300 micrograms/kg), heptylphysostigmine (5 mg/kg) and tetrahydroaminoacridine (tacrine, 5 mg/kg) on extracellular ACh levels. Baseline quantities of extracellular ACh could be detected, even in the absence of ChEI. Acetylcholine levels increased to 1100% over baseline within 30 min of physostigmine administration and returned to control levels after 1.25 hr. Heptylphysostigmine elicited a maximal increase of 1000% within 1.5 hr, and the effect persisted up to 9.5 hr. A 500% increase was observed 1.5 hr after tacrine administration, and ACh returned to control levels after 4 hr. Although the ACh effects observed in this study correlated with previously determined levels of acetylcholinesterase (AChE) inhibition, we conclude that measures of cortical AChE activity alone are not sufficient to predict extracellular ACh levels following systemic ChEI administration. PMID- 8413839 TI - Behavioural effect of pretreatment with opioid antagonists and sigma binding site ligands on the abnormal motor response produced by the kappa opioid agonist U50,488H in guinea pigs. AB - Dose-responsive motor activity induced by systemic injection of the kappa (kappa) preferring opioid agonist, U50,488H (1-10 mg/kg, s.c.) in guinea pigs was recently reported [Brent P. J. and Bot G. (1992) Psychopharmacology 107: 581 590], characterised at the higher doses used (5-10 mg/kg) by sustained postural abnormalities. The effects on the U50,488H-induced, abnormal, motor response of pharmacological manipulation of opioid receptors and sigma (sigma) sites was studied. The opioid antagonist naloxone, [5 and 15 mg/kg, subcutaneously (s.c.)], the kappa selective antagonist, norbinaltorphimine (NBNI), administered intracerebroventricularly (i.c.v., 20 and 50 nM) 0.5 hr before U50,488H, and the anticonvulsant phenytoin [25 and 50 mg/kg, intraperitoneally (i.p.)] given 1 hr before, attenuated the abnormal postures, whereas naloxone methobromide (15 mg/kg), a quaternary opioid which does not cross the blood-brain barrier, had no significant effect on the movements. In contrast, the drugs with varying affinity for sigma binding sites such as 1,3-di(2-tolyl)guanidine (DTG, 10 and 30 mg/kg), haloperidol (1 and 5 mg/kg, s.c.), dextromethorphan (1, 10 and 20 mg/kg, s.c.) and reduced haloperidol (1 mg/kg, s.c.), given 0.5-1 hr before U50,488H, exacerbated the severity of the abnormal motor activity in a dose-related manner by decreasing the latency to onset of maximum obtainable motor response and increasing the duration of the response. In addition, haloperidol (1 and 5 mg/kg, s.c.), dextromethorphan (10 mg/kg, s.c.) and DTG (30 mg/kg, s.c.), given in combination with U50,488H, induced behaviour characterised by marked oral activity. In contrast to the effect of haloperidol, pretreatment with the selective dopamine D-2 antagonist, raclopride (10 mg/kg, s.c.), had no significant effect on the abnormal movements induced by U50,488H, but did induce oral activity. These data indicate the possible involvement of kappa opioid receptors in the abnormal movement induced by U50,488H, and further demonstrate that there is an interaction between the kappa receptors and sigma sites which can influence the abnormal motor activity. PMID- 8413840 TI - Inhibitory effect of 4-phenyltetrahydroisoquinoline on locomotion and dopamine release induced by micro-injection of methamphetamine into the nucleus accumbens of the rat. AB - The inhibitory effects of 4-phenyl-1,2,3,4-tetrahydroisoquinoline (4-PTIQ) on methamphetamine-induced increases in dopamine and locomotion were investigated. Methamphetamine hydrochloride (10 micrograms) microinjected into the nucleus accumbens increased both locomotor activity and extracellular dopamine levels, measured by the brain microdialysis method. 4-PTIQ hydrochloride (20 micrograms) co-injected with methamphetamine inhibited both the increase in dopamine and the locomotor activity. The uptake blocker cocaine hydrochloride (20 micrograms) co injected with methamphetamine failed to inhibit the effect of methamphetamine. Thus 4-PTIQ but not cocaine inhibited the dopamine-releasing effect of methamphetamine, and 4-PTIQ is suggested to block methamphetamine-induced locomotion by inhibition of dopamine release. PMID- 8413841 TI - Effect of incomplete ischemia and reperfusion of the rat brain on the density and affinity of alpha-adrenergic binding sites in the cerebral cortex. Prevention of changes by stobadine and vitamin E. AB - The effect of incomplete 4 hr ischemia and subsequent 1 hr reperfusion of the rat brain on the density and affinity of alpha-adrenergic binding sites was investigated. To assess the involvement of oxygen-derived free radicals in the development of ischemic injury, we tested the effect of stobadine and vitamin E on putative changes of the binding parameters of alpha-adrenergic binding sites in ischemic and reperfused rat brains. Compared to the group of sham operated animals decreased density and increased affinity of [3H]dihydroergocryptine binding sites was found in cerebrocortical membranes of rats subjected to 4 hr incomplete ischemia and 1 hr reperfusion. The reduction of Bmax and Kd induced by ischemia and reperfusion of the brain was prevented by stobadine and vitamin E administration. Neither incomplete ischemia nor incomplete ischemia and subsequent reperfusion of the rat brain exerted significant effect on [3H]rauwolscine binding to alpha 2-adrenergic binding sites. Our results suggest that brain ischemia and reperfusion may affect the density and affinity of alpha 1- rather than of alpha 2-adrenergic binding sites. The beneficial effect of stobadine and vitamin E indicates that increased generation of oxygen free radicals might play a role in the development of these changes. PMID- 8413842 TI - Effects of minocycline on accumulation of cyclic AMP in cerebral cortex of rat. A comparison with lithium. AB - The tetracycline minocycline and lithium have been reported to share some biochemical properties. This study was aimed at investigating the effects of minocycline and lithium in vitro on accumulation of noradrenaline-, forskolin- and calcium-(Ca2+) stimulated cyclic AMP (cAMP) in the cerebral cortex of the rat. Minocycline and lithium dose-dependently inhibited noradrenaline-stimulated formation of cAMP in slices of cortex, but only lithium inhibited the formation of cAMP induced by forskolin. In contrast to lithium, minocycline did not affect either noradrenaline- or Ca(2+)-stimulated activity of adenylate cyclase in a preparation of cortical membranes. However, in slices of cortex ouabain-induced formation of cAMP (dependent on extracellular Ca2+ and blocked by the Ca2+ channel antagonist, verapamil) was reduced both by minocycline and lithium. The present results indicate that the mechanisms of action of minocycline and lithium on the cAMP signalling system in the brain of the rat differ. Minocycline does not seem to interact directly with the adenylate cyclase, as reported for lithium. The decreased agonist-stimulated production of cAMP in intact cells, in the presence of minocycline, might be due to the ability of minocycline to chelate Ca2+ ions. PMID- 8413843 TI - Resinferatoxin, an ultrapotent capsaicin analogue, has anti-emetic properties in the ferret. AB - Resinferatoxin (100 micrograms/kg, s.c.), the ultrapotent analogue of capsaicin, when given acutely blocked radiation-(200 rads) and copper sulphate (40 mg% 30 ml, p.o.)-induced emesis in ferrets and substantially decreased loperamide (0.5 mg/kg, s.c.)-induced vomiting, without significantly affecting the von Bezold Jarisch reflex or gag reflex. It also produced a decrease in core temperature as has been reported for capsaicin. The observation that resinferatoxin reduced or blocked emesis induced by both centrally (loperamide) and peripherally (CuSO4, radiation) acting stimuli suggests a novel anti-emetic action that may provide an insight into clinically useful innovative anti-emetics. The mechanism by which resinferatoxin has its anti-emetic effect is at present unknown, although the combination of results from the present study suggest a central site of action involving modulation of release of neurotransmitter, possibly in the nucleus tractus solitarius. PMID- 8413844 TI - Cocaine and dopaminergic actions in rat neostriatum. AB - Electrophysiological experiments were conducted in vivo to characterize the involvement of dopamine in effects of cocaine at the cellular level. L-DOPA (preceded by carbidopa) evoked excitation and depression equally at 0.25 mg/kg; however, at 0.5 and 1.0 mg/kg depression in the basal rate of discharge predominated. Cocaine evoked similar responses. At 0.25 and 0.5 mg/kg excitation and depression were elicited in about equal proportion, while at 1.0 mg/kg depression was predominant. Cocaine-induced neuronal changes were reversible by haloperidol. In experiments with specific dopamine antagonists, cocaine-induced neuronal depression was blocked with the D1 antagonist, SCH 23390 at a dose of 0.25 mg/kg. The D2 antagonist, eticlopride, did not alter cocaine-induced depression in any specific manner. Excitation, following cocaine, was not altered by intervention with SCH 23390, yet it was blocked with the D2 antagonist eticlopride at a dose of 0.25 mg/kg. The duration of these changes was consistent with the observed duration of the psychotropic actions of cocaine. PMID- 8413846 TI - Current bibliographies of neuropeptides prepared by the University of Sheffield Biomedical Information Service. PMID- 8413845 TI - Pharmacological characterization of the release of neuropeptide Y-like immunoreactivity from the rat hypothalamus. AB - Radioimmunoassay for NPY and detection by HPLC-EC were used to detect the concurrent release of norepinephrine and NPY from slices of the rat hypothalamus and to examine the modulation by adrenoceptors of the release of this neuropeptide. Basal and potassium-evoked (56 mM K+) release of both compounds were easily measured, with evoked release occurring in a calcium-dependent manner. The effect of the alpha 2-adrenoceptor agonist clonidine, the antagonists prazosin (alpha 1 selective) and yohimbine (alpha 2 selective) and the beta adrenoceptor antagonist propranolol were all shown to modulate the evoked release of NPY. The alpha 2 agonist clonidine decreased evoked release of NPY, while the alpha 2 antagonist yohimbine increased the potassium-evoked release. Prazosin decreased both the basal and potassium-evoked release of NPY. Propranolol had the most profound effect on release of NPY, causing a significant decrease in basal release and a large decrease in the potassium-evoked release of NPY from slices of hypothalamus. PMID- 8413847 TI - Structure and functional expression of a complementary DNA for porcine growth hormone-releasing hormone receptor. AB - Growth hormone-releasing hormone (GHRH) belongs to the family of gut-neuropeptide hormones which also includes glucagon, secretin and vasoactive intestinal peptide (VIP). All receptors for this peptide hormone family seem to involve similar signal transduction pathways. Upon hormone binding, these receptors interact with guanine nucleotide binding protein 'Gs' and cause the stimulation of adenylate cyclase. The secretin and VIP receptor cDNAs have recently been cloned and found to be homologous to those of calcitonin and parathyroid hormone receptors. Based on cDNA sequences of these receptors, we designed several oligonucleotide primers which were used to amplify two novel porcine pituitary cDNA fragments by the polymerase chain reaction. One novel receptor cDNA fragment was used to screen a porcine pituitary cDNA library and a full-length cDNA encoding a putative porcine GHRH receptor of 451 amino acids was isolated. This putative receptor mRNA is present specifically in porcine anterior pituitary cells and not in eight other porcine tissues as shown by Northern hybridization analysis. The receptor cDNA was subsequently cloned into a mammalian cell expression vector containing the cytomegalovirus promoter. A human kidney tumor cell line (293) stably transfected with this vector was found to express the receptor efficiently and to bind [125I] GHRH specifically. Furthermore, challenge of the 293 cells expressing the receptor by GHRH leads to efficient stimulation of cytoplasmic cAMP production. PMID- 8413848 TI - Distribution and cellular localization of vasopressin mRNA in the ovine brain, pituitary and pineal glands. AB - In this study, in situ hybridization histochemistry was used to determine the regional and cellular localization of vasopressin-neurophysin II (AVP) mRNA in the sheep brain and pituitary with an 35S-labelled synthetic 45-mer oligonucleotide probe complementary to the bovine AVP gene. The highest densities of labelled cell bodies were found in the paraventricular nucleus (PVN), supraoptic nucleus (SON) and suprachiasmatic nucleus (SCN) of the hypothalamus, though such cells were also found in other regions of the diencephalon, including the accessory magnocellular nuclei. Labelled cells were also observed sparsely distributed in every major cortical field as well as in choroid plexus and the pineal gland. No AVP mRNA-expressing cells were found in the bed nucleus of the stria terminalis, the amygdala, or in the medulla and brainstem. In the pituitary, a dense AVP mRNA signal was observed in the intermediate lobe whereas, cells in the anterior or neural lobe did not express AVP mRNA. The dense population of AVP-expressing neurons in both magnocellular and parvocellular fields of the hypothalamus support major roles of AVP in both posterior and anterior pituitary function. Finally, the extrahypothalamic distribution of AVP mRNA transcripts suggest that vasopressinergic neurons may be involved in diverse physiological functions, including the regulation of pineal function and cognition. PMID- 8413849 TI - Processing, release and metabolism of cholecystokinin in SK-N-MCIXC cells. AB - The human cholinergic neuroepithelioma cell line SK-N-MCIXC, which expresses high levels of cholecystokinin (CCK) mRNA and secretes intact CCK into the media, was used to examine CCK processing and metabolism. Our data provide evidence for the existence of specific candidate processing enzymes in SK-N-MCIXC cells which may be involved in processing proCCK in the brain and indicate that SK-N-MCIXC cells provide a model system for studying the regulation of these enzymes. mRNAs for the intracellular processing enzymes, prohormone convertase 1 (PC1), PC2 and furin were present in SK-N-MCIXC cells. PC1 and/or PC2 and/or furin may cleave at the dibasic amino acid pairs Arg-Arg at the C-terminal part of proCCK, and Arg-X X-Arg at the N-terminal of the CCK-58 sequence in proCCK. The SK-N-MCIXC cell line demonstrated spontaneous and regulated release of CCK and large amounts of CCK-precursors, as measured with region specific radioimmunoassays coupled to high performance liquid chromatography. Storage granules containing glycine extended CCK were shown in SK-N-MCIXC cells using indirect immunofluorescence. The extracellularly localized CCK-metabolizing enzyme, neutral endopeptidase 24.11 (EC 3.4.24.11), was present in membranes from both SK-N-MCIXC cells and in intact slices of rat cerebral cortex. The rat cerebral cortex is a brain region known to be rich in CCK. The SK-N-MCIXC cell line provides an in vitro model to study the regulation of CCK synthesis and metabolism in neuronal systems since it contains the storage granules, mRNA, intact peptide, and complement of enzymes necessary for biosynthesis and metabolism of CCK. PMID- 8413850 TI - The evidence of thyroliberin/triiodothyronine control of TSH secretory response from human peripheral blood monocytes cultured in vitro. AB - Immune system function has been shown to be under the influence of various neuromodulators and endocrine system peptides. This in vitro study describes the stimulatory effect of thyrotropin releasing hormone (thyroliberin, TRH) on thyrotropin (TSH) release from cultured human peripheral blood monocytes. The stimulatory effect of TRH on TSH release from monocytes is totally blocked by triiodothyronine (T3) administrations. These results indicate that TSH release from human monocytes is under the control of TRH/T3 mechanisms, similar to hypothalamic-pituitary-thyroid axis. PMID- 8413851 TI - Nitric oxide is not involved in the effects induced by non-adrenergic non cholinergic stimulation and calcitonin gene-related peptide in the rat mesenteric vascular bed. AB - The mechanism involved in the effects induced by the activation of perineural non adrenergic non-cholinergic (NANC) nerves or by exogenous calcitonin gene-related peptide (CGRP) was investigated in the rat mesenteric vascular bed (MVB) perfused with Kreb's solution containing methoxamine and guanethidine. The activation of NANC terminals of the tissue was carried out by means of electrical field stimulation (EFS). An increase in the perfusion pressure of the preparations was observed in the presence of two inhibitors of nitric oxide synthase: NG monomethyl-L-arginine (L-NMMA) (100 microM) and NG-nitro-L-arginine methyl ester (L-NAME) (100 microM). However L-NMMA and L-NAME did not modify the relaxant effect induced by EFS and exogenous CGRP. Furthermore the relaxant effect induced by EFS and exogenous CGRP was not affected by the removal of endothelium from the preparations. These results provide evidence that the vasodilation induced by NANC stimulation or by exogenous CGRP in MVB does not involve NO production. PMID- 8413852 TI - Frog prodermorphin expressed in mammalian cells is partly converted to the hydroxyproline containing precursor. AB - Using recombinant vaccinia virus, we have expressed in mammalian cells the cDNA coding for the precursor of dermorphin, a D-alanine containing opioid peptide from the skin of the South American frog Phyllomedusa sauvagei. HeLa cells and AtT-20 cells produced prodermorphin where proline-6 of dermorphin was partly hydroxylated. This was demonstrated by digesting the partially purified precursors with trypsin and carboxypeptidase B. After immunoprecipitation and separation by HPLC, two decapeptides were detected which differed by the presence of proline or hydroxy-proline at position 6. This demonstrates that HeLa cells as well as AtT-20 cells can perform the post-translational conversion of certain proline residues to hydroxyproline in a foreign hormone precursor expressed in these cells. PMID- 8413853 TI - Effect of CCK-8 on audiogenic epileptic seizure in P77PMC rats. AB - P77PMC rat is a breed of rat with congenital audiogenic seizure(AS). AS attacks were suppressed by cholecystokinin octapeptide (CCK-8) injected intraperitoneally (i.p.) at a dose of 50 micrograms/kg, but not at 25 micrograms/kg. Radioimmunoassay study showed that the CCK-8 immunoreactivity (IR) in the cerebrocortex and hippocampus is much lower in P77PMC rats than that of Wistar rats. The results suggest that a low cerebral content of CCK-8 may account for the high susceptibility of audiogenic seizure in P77PMC rats. PMID- 8413854 TI - Current bibliographies of neuropeptides prepared by the University of Sheffield Biomedical Information Service. PMID- 8413855 TI - Neurokinin A-like immunoreactivity in the cat brainstem. AB - We have studied the distribution of neurokinin A-like immunoreactive cell bodies and fibers in the cat brainstem. The densest clusters of perikarya containing the peptide were observed in the periaqueductal gray, inferior colliculus, postpyramidal nucleus of the raphe, medial nucleus of the solitary tract and in the lateral reticular nucleus. By contrast, the interpeduncular nucleus, magnocellular part of the red nucleus, central tegmental field, cuneiform nucleus, dorsal tegmental nucleus, nucleus sagulum and the medial and inferior vestibular nuclei had the lowest density, whereas a moderate density of immunoreactive cell bodies was found in the superior colliculus, medial division of the dorsal nucleus of the raphe, nucleus incertus, locus coeruleus and in the Kolliker-Fuse area. The highest density of immunoreactive fibers was observed in the substantia nigra, periaqueductal gray, marginal nucleus of the brachium conjunctivum, medial vestibular nucleus, medial nucleus of the solitary tract, laminar spinal trigeminal nucleus, inferior colliculus, medial division of the dorsal nucleus of the raphe, locus coeruleus, dorsal tegmental nucleus and in the spinal trigeminal tract. A moderate density of immunoreactive fibers was found in the dorsal motor nucleus of the vagus and in the postpyramidal nucleus of the raphe and a low density in the cuneiform nucleus, Kolliker-Fuse area, nucleus sagulum, inferior and superior central nuclei, lateral reticular nucleus and in the lateral and magnocellular tegmental fields. PMID- 8413856 TI - Failure of intravenous pentagastrin challenge to induce panic-like effects in rhesus monkeys. AB - We examined the ability of intravenous (i.v.) challenge with pentagastrin to induce behavioural and cardiovascular effects consistent with panic attack in conscious rhesus monkeys. For behavioural evaluation, 4 naive male rhesus monkeys familiar with minimal manual restraint necessary for drug administration received a rapid i.v. bolus of pentagastrin (4, 8 or 16 micrograms/kg) or water on four separate occasions according to a randomised cross-over design. Behaviour was rated by a blind observer continuously during, and for the first 5 min immediately following i.v. injections while the monkey sat on the handler's lap, and then for a further 25 min in an individual observation cage. In separate experiments, the ability of pentagastrin to alter cardiovascular parameters which may accompany panic or anxiety (elevated heart rate and blood pressure) was explored. For cardiovascular studies, 8 male or female rhesus monkeys with femoral artery catheters were chair restrained and received a bolus injection of pentagastrin (4, 8 or 16 micrograms/kg) or saline into the saphenous vein at 30 min intervals. Blood pressure and heart rate were monitored continuously using a Statham Gould pressure transducer. Pentagastrin induced no consistent behavioural or cardiovascular changes. Similar pilot studies using CCK4 also failed to reveal such effects. We conclude that CCK-induced panic-like effects may not be demonstrable following challenge with pentagastrin under laboratory conditions in rhesus monkeys. PMID- 8413857 TI - Kinetic parameters of antagonism by the delta opioid receptor selective peptide antagonist Boc-Tyr-Pro-Gly-Phe-Leu-Thr against selective and non-selective agonists in the mouse vas deferens. AB - The kinetic parameters of antagonism by the delta opioid receptor selective antagonist N-t-Boc-Tyr-Pro-Gly-Phe-Leu-Thr, obtained by using moderately selective or selective agonists, were compared in the mouse vas deferens bioassay. The apparent affinity for the preferred receptor type was 6.8 times higher when selective agonist was used, resulting in a Ke of 81.4 nM (66.3-99.9, n = 6) against [D-Ala2, D-Leu5]-enkephalin, with a 3700-fold delta over mu or kappa selectivity ratio. PMID- 8413858 TI - Differential processing of provasotocin: relative increase of hydrin 2 (vasotocinyl-Gly) in amphibians able to adapt to an arid environment. AB - Hydrin 2 (vasotocinyl-Gly) is an intermediate in pro-vasotocin processing found, along with vasotocin, only in the neurohypophysis of anuran amphibians. It increases cutaneous water permeability in the frog and is likely involved in neuroendocrine control of osmoregulation. The relative amounts of vasotocin and hydrin 2 stored in neurohypophysis have been measured on the one hand in amphibian species known not to adapt in dry areas, on the other hand in two species, Bufo regularis (Africa) and Bufo viridis (Near-East) able to survive in an arid environment. In the first group, the proportions of the two peptides are approximately equal whereas in the two toads the molar ratio hydrin 2 to vasotocin reaches 2. The ratio does not appear to vary significantly when these toads are either submitted to dehydration or placed in saline solutions. Predominance of hydrin 2 suggests an adaptive decrease of the activity of the alpha-amidating enzymatic system involved in the conversion of vasotocinyl-Gly into mature amidated vasotocin. PMID- 8413859 TI - Unusual effect of prolonged vasoactive intestinal peptide (VIP) administration on the adrenal growth and corticosterone secretion in the rat. AB - VIP is widely distributed in the body, and among others is present within the hypothalamo-pituitary-adrenal axis. The aim of this study was to investigate the effect of prolonged VIP administration on the structure and function of the rat adrenal cortex. Adult female rats were treated for 7 or 14 days with VIP (1.5 micrograms/100 g/day). This treatment did not change adrenal weight and evoked only slight changes in the stereologic parameters of adrenal cortex (volume of adrenocortical zones, average cell volume and number of cells in the gland). On the contrary, basal plasma corticosterone levels were markedly lowered (44% and 26% of control values in 7- and 14-day experiments, respectively), an effect accompanied by a significant decrease in corticosterone output by adrenal slices. After standardized ether stress, blood corticosterone concentration was similar in control and 14-day VIP-treated rats, while corticosterone secretion by adrenal quarters was higher in VIP-administered animals. Thus, prolonged VIP administration results in an unusual inhibition of corticosterone secretion in the rat, which is not coupled with a compensatory hypertrophy of the rat adrenal cortex. Such changes may depend on a modulatory action of VIP on the hypothalamo pituitary-adrenal axis, and its possible mechanism is discussed. PMID- 8413860 TI - Release of the prohormone convertase PC1 from AtT-20 cells. AB - AtT-20 cells are known to synthesize two molecular weight forms of the prohormone converting enzyme PC1 with molecular masses of 87 and 66 kDa. In this study we have analyzed basal and stimulated secretion of these proteins. Western blot results show that basal secretion medium of cultured AtT-20 cells contained low concentrations of both the 87 and 66 kDa forms of PC1 with the former protein predominant. During the stimulation period with CRF, cAMP and cAMP + BaCl2, increased release of both proteins was observed, but the 66 kDa protein predominated. Secretion medium obtained from stimulated and unstimulated cells was enzymatically active against the Cbz-Arg-Ser-Lys-Arg-AMC fluorogenic substrate as well as against 35S-proenkephalin. This activity was Ca+2 dependent and was inhibited by the chelating agent EDTA. The activity was insensitive to acid and thiol proteinase inhibitors as well as to N-alpha-p-tosyl-L-Lys chloromethyl ketone; it was slightly sensitive to phenylmethyl sulfonyl fluoride and was strongly inhibited by D-Tyr-Ala-Lys-Arg-chloromethyl ketone. This inhibitor profile exhibits strong similarities to furin and kexin. After partial purification of medium by gel filtration chromatography, a portion of the enzymatic activity and immunoreactivity for both 87 kDa and 66 kDa proteins eluted with an apparent molecular weight of 400 kDa (suggesting aggregation); however the highest activity appeared in the elution position of the 66 kDa monomer. When the 87 kDa protein was removed from the medium by means of an affinity column containing an antibody against the carboxyl terminal portion of PC1, the column flow-through, which included the 66 kDa protein, still remained enzymatically active. These data support the notion that the 66 kDa protein, which is the most concentrated PC1 product stored in AtT-20 cells and is released during stimulation, is enzymatically active. PMID- 8413861 TI - Immunoreactivity to vasoactive intestinal polypeptide (VIP) in the hypothalamus of the domestic fowl, Gallus domesticus. AB - The distribution of VIP-immunoreactive neurons and fibers was detected in the hypothalamus of the chick by immunohistochemistry and light microscopy. A large amount of VIP cellular bodies was localized in the anterior and medial area of the hypothalamus with the highest density of cells in supraoptic, magnocellular preoptic, suprachiasmatic and paraventricular nuclei. Only few VIP-immunoreactive neurons were observed in the caudal section of infundibuli nucleus. A considerable concentration of VIP-positive fibers was also detected in the external layer of the anterior and posterior median eminence. Their presence might have origin both from the neurons of the infundibuli nucleus and from the cells of the paraventricular nucleus. Few VIP-immunoreactive fibers were revealed in the organum vasculosum of the lamina terminalis. These results tend to suggest that VIP may play more than one role in the hypothalamic regions, particularly in the preoptico-hypothalamic area. The presence of this peptide in the median eminence supports even more the hypothesis that it may be released into the portal circulation and transported to the pars distalis of the pituitary gland. PMID- 8413862 TI - Opioid regulation of proopiomelanocortin (POMC) gene expression in the rat brain as studied by in situ hybridization. AB - Proopiomelanocortin (POMC) is the precursor of the potent opioid peptide beta endorphin as well as a number of other active peptides. On the basis of neuroanatomical data indicating the presence of contacts between POMC neurons in the rat arcuate nucleus, it has been proposed that POMC neurons could be autoregulated. In order to investigate the role of opiates in the regulation of POMC gene expression in the rat arcuate nucleus, we studied the effects of chronic administration of the opioid drug morphine and an opiate receptor antagonist naloxone on POMC mRNA levels as measured by in situ hybridization, 4 day treatment with naloxone (4 mg/kg/day) produced a 60% increase in the number of silver grains overlying POMC neurons. Conversely, morphine (40 mg/kg/day) also administered during 4 days decreased the hybridization signal by 30%. The concomitant administration of morphine and naloxone completely prevented the effect of morphine on POMC gene expression indicating that the inhibitory influence of morphine is likely to be mediated by opioid receptors. The data obtained clearly indicate that activation of opioid receptors decreased the biosynthetic activity of POMC neurons and that conversely opiate receptor blockade caused an increase in the activity of these neurons. They are consistent with the hypothesis of an autoregulation of the POMC neuronal system by endogenous opiate peptide(s). PMID- 8413863 TI - Renal osteodystrophy of the cervical spine: neurosurgical implications. AB - Renal osteodystrophy describes the changes in bone physiology seen in renal failure. Traditionally, these changes resulted in mild neurological sequelae that were effectively managed medically. Despite the often severe destruction seen on x-ray, surgical therapy has not been reported extensively. With increasing numbers of long-term hemodialysis survivors, however, a newly recognized erosive spondyloarthropathy with extensive bony destruction is seen more frequently. These changes can mimic the radiographic appearance of osteomyelitis, not an uncommon sequela of hemodialysis patients; therefore, this diagnosis must be considered. Although renal osteodystrophy can be seen throughout the spine, reports of significant cervical spine involvement are uncommon. We present our experience with six cases of renal osteodystrophy of the cervical spine requiring surgical intervention for instability or cord compression. Osteomyelitis was present in two of the patients. The pathophysiology and radiographic characteristics of renal osteodystrophy and the incidence of osteomyelitis are discussed. Fusion techniques, including the use of anterior and posterior internal and external stabilization, are presented. These patients represent a therapeutic challenge for the neurosurgeon, given the underlying bone pathology. PMID- 8413864 TI - Posterior cervical laminoforaminotomy for radiculopathy: review of 172 cases. AB - Laminoforaminotomy performed with the patient in the sitting position with our improved techniques represents an effective treatment for cervical radiculopathy. We present the results of laminoforaminotomies performed in 172 patients with cervical radiculopathy during a 7-year period. The posterior approach in the surgical management of cervical radiculopathy is not only acceptable, but in certain cases is preferable to the anterior approach. When the abnormality is central, broad based and anterior, posterior procedures are unlikely to achieve decompression. However, with lateral or foraminal nerve root compression, the simpler posterior keyhole laminoforaminotomy works well. In our opinion, physicians advocating either procedure exclusively are not providing the patient with the optimal level of care. Our purpose is to present in detail our surgical technique in conjunction with an analysis of our long-term results in clinical situations in which our technique is clearly indicated. PMID- 8413865 TI - Cervical myelomeningoceles. AB - Cervical myelomeningoceles are rare dysraphic lesions. Nine cases of cervical myelomeningoceles are reported. The external features of all nine myelomeningoceles were strikingly similar: They were sturdy, tubular protuberances from the back of the infants' necks, covered at the base by full thickness skin and covered on the dome by thick squamous epithelium. Internally, these were tethered cord lesions in which fibroneural bands or sagittal midline fibrous septa were tightly tethering the cervical spinal cord to the adjacent dural or intrasaccular soft tissues. Six of our early cases (Group 1) were initially treated with simple subcutaneous resection of the sac and ligation of the dural fistula without release of the internal tethering structures. Five of these children subsequently deteriorated 13 months to 8 years later, all with worsening hand function and spastic legs. All five were reexplored, and the tethering bands and septa were excised; all showed improvement. The other three neonates (Group 2) treated in the last 4 years underwent initial intradural exploration of the lesions; in one case, the tethering fibrous elements were only partially eliminated and the patient deteriorated 4 years later, but improved after a second operation for resection of a missed ventral fibrous septum. The other two Group 2 infants had a thorough release of the fibroneural stalks initially, and both were neurologically stable 3 years later. We recommend that cervical myelomeningoceles should be studied preoperatively with magnetic resonance imaging and computed tomographic myelography to identify the internal structures. The minimum initial surgical treatment should be a two-level laminectomy, intradural exploration, and excision of all tethering bands and septa, in addition to resection of the sac. If a split cord is revealed by imaging studies, both the ventral and dorsal surfaces of the hemicords must be carefully inspected to locate the median septum. PMID- 8413866 TI - Tethered cord syndrome and the conus in a normal position. AB - The intent of this article is to propose the existence of the tethered cord syndrome in patients whose conus is in a normal position. The tethered cord syndrome has been a well-recognized entity, occurring in the pediatric and adult population. A central tenet to this syndrome is that the conus medullaris must be "abnormally" low, regardless of the tethering lesion. Our 12-year series of 73 patients with tethered cord syndrome included 13 patients in whom the cord terminated at or above the L1-L2 space. These patients otherwise displayed characteristics usually associated with the patient with an abnormally low conus. Most patients with progressive neurological deterioration responded to surgery. The preoperative radiographic evaluation and the operative findings usually demonstrated a thickened filum. Tethered cord syndrome may occur in the patient who has a conus in the "normal" position. PMID- 8413867 TI - Pain, disability, and psychological functioning in chronic low back pain subgroups: myofascial versus herniated disc syndrome. AB - A series of patients with chronic low back pain evaluated at a tertiary referral center were the subjects for this study. Of 250 consecutive patients, 94 were diagnosed as having myofascial pain and 57 as having herniated disc syndrome. Before evaluation and diagnosis, all patients completed the McGill Pain Questionnaire, ratings of pain and disability, and the Symptom Checklist 90-R. Patients were also grouped on the basis of previous surgical history and workers' compensation benefits. Patients suffering from myofascial pain were significantly less likely to report periods of pain relief than patients with herniated disc syndrome. Those receiving workers' compensation benefits reported significantly greater levels of pain, disability, and psychological distress than those not receiving benefits, irrespective of diagnosis. Patients who underwent previous surgery did not differ significantly from those who never underwent surgery. All patients had elevated scores on the Somatization subscale of the Symptom Checklist 90-R. Patients with myofascial pain and workers' compensation benefits demonstrated the highest levels of somatization and phobia. These findings suggest that the effects of low back pain of myofascial origin have comparable, if not worse, consequences than disc herniation. These findings also reaffirm the importance of workers' compensation in understanding the differences in patients with chronic low back pain. PMID- 8413868 TI - Changes in cerebral blood flow and metabolism related to the presence of subdural hematoma. AB - Acute subdural hematoma (SDH) remains an important factor in head injury. The early effects of SDH on cerebral blood flow (CBF) and cerebral metabolic rate of oxygen consumption (CMRO2) in humans have not been clearly demonstrated. Patients admitted to the Medical College of Virginia with severe closed-head injury between 1982 and 1990 were studied with Xenon-133 regional CBF measurement. Data were reviewed retrospectively with regard to the presence of SDH (n = 54). A comparison group consisted of patients with head injuries without mass lesions or midline shift on admission computed tomographic scans (n = 76). CBF measurements made in patients less than 16 years of age, with concurrent administrations of mannitol or vasopressors, or with cerebral perfusion pressure under 50 mm Hg were excluded. CBF measurements were made on multiple occasions during the first 6 days after injury, and in many instances, simultaneous determinations of cerebral arteriovenous oxygen difference (AVDO2) were made through sampling of jugular bulb and arterial oxygen content. Not all patients underwent CBF measurements on each day. Differences in mean CBF, CMRO2, and AVDO2 were evaluated on each day after injury with the application of Student's t-test for independent groups. Significant reductions in CBF were demonstrated in patients with SDH on Days 1 (P < 0.0005) and 2 (P < 0.01). CMRO2 differed notably on Days 1 (P < 0.005) and 2 (P < 0.05) in patients with SDH, but when corrected for the lower Glasgow Coma Score in patients with SDH, the P values were only 0.07 and 0.12, respectively (analysis of covariance).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413869 TI - Grade zero removal of supratentorial convexity meningiomas. AB - Although meningiomas are benign intracranial tumors, their frequency of recurrence after surgery has not been as low as expected. The recurrence rate of meningiomas is clearly related to the degree of tumor removal. Simpson Grade I removal, which entails excising the tumor and its dural and sinus attachments, is associated with the lowest rate of recurrence. To further minimize the recurrence of convexity meningiomas, we removed an additional dural margin of about 2 cm around the tumor (Grade 0 removal). For tumors involving bone, we removed the hyperostotic bone with a healthy margin and pericranium in en bloc resection. Between 1982 and 1992, 37 patients (15 men, 22 women) with an average age of 52.1 years were operated on by the above technique. Nineteen had a follow-up period of more than 5 years. To date, no tumors have recurred and no morbid incidences have occurred with this maneuver. We believe that the recurrence rate of convexity meningiomas can be diminished by including in the resection a margin of dura that might harbor a foci of tumor cells. PMID- 8413870 TI - Perinidal hypervascular network on immediate postoperative angiogram after removal of large arteriovenous malformations located distant from the arterial circle of Willis. AB - Unexplained intraoperative or postoperative hemorrhage occurs in certain cases in the surgery of large arteriovenous malformations (AVMs), which are defined as those with maximal diameter of the nidus larger than 5 cm. Immediate postoperative angiograms were evaluated with reference to whether such hemorrhage could be predicted. Immediate postoperative angiograms were taken in 12 patients with large AVMs on the day after the operation during which the AVM was removed. In five of six AVMs located in the distal cerebral hemisphere, a hypervascular network was seen in the vicinity of the removed nidus on the immediate postoperative angiogram. The unexplained hemorrhage occurred in four of the five cases with the hypervascular network. In the remaining six patients with AVMs, the hypervascular network was not observed and they did not develop hemorrhage; the AVMs were located either close to the arterial circle of Willis or in the cerebellum. We believe that the hypervascular network seen on the immediate postoperative angiogram after the removal of a large distal AVM is an important sign predicting the likelihood of unexplained intraoperative and postoperative hemorrhage development. We call this perinidal hypervascular network "modja-modja vessels" because it resembles shaggy hair. The mechanism of its appearance and the pathogenesis of the unexplained intraoperative and postoperative hemorrhage are discussed. PMID- 8413871 TI - Effects of four intravenous anesthetic agents on motor evoked potentials elicited by magnetic transcranial stimulation. AB - The influence of four intravenous anesthetic agents on motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation (magnetic MEP) was examined in 77 subjects. The patients were anesthetized by a continuous intravenous infusion of one of the following anesthetic agents: propofol, etomidate, methohexital, or thiopental. Comparable anesthetic effects among the four agents were achieved by computing an infusion scheme for each drug. Infusion rates were increased slowly in a step-wise manner in order to reach minimal anesthetic blood concentrations within 15 minutes. During anesthesia induction, magnetic MEPs were recorded every 2 minutes from the abductor pollicis brevis muscle. The patient's level of consciousness was assessed and documented in the alternating minutes. A dose-related reduction of the MEP amplitudes was seen in all drug groups, while the latencies remained constant. Reduction of the amplitude was occasionally so prominent that the MEP was completely abolished before adequate anesthesia was achieved. MEPs were obtainable at the end of anesthesia induction in 14% of the propofol group (n = 22), 57% of the etomidate group (n = 21), 53% of the methohexital group (n = 19), and 20% of the thiopental group (n = 15). Propofol and thiopental showed significantly stronger suppression of MEP, when compared to etomidate (both P < 0.01) and to methohexital (P = 0.01 and 0.05, respectively). Etomidate was the least detrimental anesthetic agent for intraoperative monitoring of magnetic MEP. Nonetheless, the low incidence of 57% of preserved MEP in subjects without motor deficits indicated the inadequacy of this technique for intraoperative monitoring. More effective transcranial stimulation techniques are required for successful intraoperative MEP monitoring. PMID- 8413872 TI - Ceftizoxime versus vancomycin and gentamicin in neurosurgical prophylaxis: a randomized, prospective, blinded clinical study. AB - In a prospective, randomized, blinded study, 826 patients undergoing clean neurosurgical procedures received single intravenous doses of ceftizoxime (2 g) (n = 422) or a combination of vancomycin (1 g) and gentamicin (80 mg) (n = 404) 1 hour before an incision was made. Patients with infected or contaminated wounds and those receiving shunts or other implants were excluded. Primary wound infections occurred within 30 days in five patients in each group and were most common after spinal surgery and procedures through previous incisions. Secondary infections (pneumonias, urinary tract infections, and intravenous line-related bacteremia) occurred in 24 patients in the ceftizoxime group and 25 in the vancomycin/gentamicin group. The infection rates after transsphenoidal procedures (n = 129) were remarkably low in both groups. Ceftizoxime caused no adverse drug reactions, but six patients in the vancomycin/gentamicin group had clinically significant infusion-related hypotension or flushing. Placement of a temporary external drain, use of an operating microscope, preoperative steroids, and diabetes were not associated with increased infection rates. Analysis of routinely encountered ventricular cerebrospinal fluid and simultaneously obtained peripheral blood showed low but detectable levels of all three antibiotics within 2 hours; only ceftizoxime, however, achieved cerebrospinal fluid levels sufficient to inhibit the staphylococcus and Gram-negative bacilli most often associated with postneurosurgical infections. We conclude that ceftizoxime is as effective as vancomycin and gentamicin in neurosurgical prophylaxis but is less toxic and penetrates cerebrospinal fluid better. PMID- 8413873 TI - Duration of intracranial pressure monitoring does not predict daily risk of infectious complications. AB - A group of 205 patients (115 children and 90 adults) with a total of 212 intracranial pressure (ICP) monitors were retrospectively studied with attention to daily cerebrospinal fluid cultures, duration of monitoring, associated cranial injuries, and hospital site of the ICP monitor (intensive care unit or operating room). Only closed ICP monitoring systems without irrigation or compliance testing were used, and all patients received antibiotics as prophylaxis throughout the monitoring period. There were no complications associated with monitor placement. Incidence histograms and regression analysis were used to determine the daily risk of subsequent infections, in addition to evaluating the cumulative risk of infection, as has been previously described in the literature. No relation between the duration of ICP monitoring and the rate of daily infection through the period of maximal monitoring (1-2 weeks) was found in this series. The overall incidence of infection was 7.1% with a median duration of monitoring of 7.2 days. The age of the patient (adult vs. child), site of ICP monitor placement, and nature of the underlying disease (trauma vs. nontrauma) had no significant effect on the development of monitor-related infections in our study. These data indicate that the decision to continue ICP monitoring can be based solely on the clinical necessity for further monitoring rather than on concerns for monitor removal to prevent infection. PMID- 8413874 TI - Proof of equivalence. The inference of statistical significance. "Caveat emptor". PMID- 8413875 TI - Permissible temporary occlusion time in aneurysm surgery as evaluated by evoked potential monitoring. AB - To clarify the permissible time for temporary vascular occlusion during aneurysm surgery, we have undertaken a retrospective analysis of the results of intraoperative median nerve somatosensory evoked potential (SEP) monitoring in 97 patients with middle cerebral artery (MCA) (46 patients) or internal carotid artery (ICA) (51 patients) aneurysms. All patients underwent temporary vascular occlusion, lasting from 2 to 70 minutes, under the administration of a mixed solution of 20% mannitol with phenytoin and vitamin E. The mean occlusion time was 20.3 minutes for the MCA patients and 15.8 minutes for the ICA patients. The SEP disappeared during occlusion in 42 patients (30 MCA and 12 ICA). All but three eventually recovered the SEP to the baseline level after recirculation, and none of the 39 patients had postoperative sequelae. The time period from the start of occlusion until the complete loss of the SEP averaged 8.6 minutes among these 39 patients, and the occlusion time from total SEP loss until recirculation averaged 12 minutes. In the remaining 3 of the 42 patients (all 3 being ICA aneurysm patients), however, the SEP did not recover after recirculation and all 3 patients showed postoperative sequelae. In two of these three patients, vascular occlusions were performed at the multiple sites (i.e., at the ICA, MCA, anterior cerebral artery, and posterior communicating artery) and the SEP disappeared rapidly after the occlusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413876 TI - Variations in regional cerebral blood flow investigated by single photon emission computed tomography with technetium-99m-d, l-hexamethylpropyleneamineoxime = l-h during temporary clipping in intracranial aneurysm surgery: preliminary results. AB - Single photon emission computed tomography with technetium-99m-d, l hexamethylpropyleneamineoxime was used to assess variations in regional cerebral blood flow during temporary clipping in the course of intracranial aneurysm surgery and during the postoperative period in 20 patients, 14 of whom underwent temporary clipping. Of these 14 patients (Group A), 9 had aneurysms of the anterior communicating artery, 2 had aneurysms of the middle cerebral artery, and 3 had aneurysms of the carotid siphon. Temporary clips were applied, according to the site of the lesion, on A1, on the trunk of the middle cerebral artery, or on the trunk of the internal carotid artery. The occlusion time ranged from 2 to 31 minutes. The six patients who did not undergo temporary clipping served as controls (Group B), as follows: three had aneurysms of the posterior communicating artery, one of the anterior communicating artery, one of the middle cerebral artery, and one of the internal carotid artery. All patients were investigated with cerebral single photon emission computed tomography preoperatively, perioperatively, and postoperatively. In all the patients of Group A, the preliminary results of the study show a sharp fall in the perfusion of the territories of the temporarily clipped parent vessel and practically a complete recovery within 2 to 7 days of surgery, with no significant neurological symptoms. No similar disturbance of perfusion was found in the patients of Group B. PMID- 8413877 TI - Spinal cord evoked potential monitoring after spinal cord stimulation during surgery of spinal cord tumors. AB - Spinal cord evoked potentials (SCEPs) after spinal cord stimulation were used as a method of spinal cord monitoring during surgery of 6 extramedullary and 14 intramedullary spinal cord tumors. SCEPs were recorded from an epidural electrode placed rostral to the level of the tumor. Electrical stimulation was applied on the dorsal spinal cord from a caudally placed epidural electrode. The wave forms of SCEPs consisted of a sharp negative peak (N1) in 15 cases and two negative peaks (N1 and N2) in 5 cases. The N2 wave was markedly attenuated by posterior midline myelotomy, whereas the N1 activity showed less-remarkable changes by myelotomy. An increase in N1 amplitude was observed after the removal of the tumor in four extramedullary and three intramedullary cases. Of six patients that showed decreased N1 amplitude after the removal of the tumor, five patients developed postoperative motor deficits. However, there were four false-negative cases and one false-positive case in regard to changes of N1 amplitude and postoperative motor deficits. Four false results occurred in intramedullary cases. In two of them, postoperative symptoms indicated intraoperative unilateral damage to the spinal cord. The position of the stimulating electrode, the difference in thresholds of the axons for electrical stimulation between the right and left side of the spinal cord, or the change of the distance between the electrode and the spinal cord surface may account for these false results. Thus, our analysis of the changes of SCEP wave forms and early postoperative symptoms indicates that the sensitivity of this monitoring method to detect intraoperative insults to the spinal cord is unsatisfactory in spite of the reproducible wave forms. We conclude that SCEP monitoring can be used as an alternative method or in combination with other types of evoked potentials in patients with severe spinal cord lesions who show abnormal somatosensory evoked potentials preoperatively. PMID- 8413878 TI - Transpetrosal approach: surgical anatomy and technique. AB - Transpetrosal operations have been shown to offer distinct advantages over traditional operations in approaching lesions of the petroclival area. Confusion about these approaches exists due to the variety of names given to these procedures and the lack of detailed descriptions needed to perform them. After extensive review of the literature, we have determined that all transpetrosal techniques fall into one of two categories: anterior petrosectomy or posterior petrosectomy. Combining one of these procedures with existing conventional procedures accurately describes all existing transpetrosal operations and eliminates confusion over nomenclature. In addition, through a series of cadaveric dissections and operative experience, we have detailed each of these procedures as a series of steps that will enable the surgeon to understand the unfamiliar anatomy of the temporal bone and to perform these transpetrosal techniques. PMID- 8413879 TI - Spinal cord blood flow and evoked potential responses after treatment with nimodipine or methylprednisolone in spinal cord-injured rats. AB - This study examined the effect of nimodipine or methylprednisolone on spinal cord blood flow (SCBF) and electrophysiological function after spinal cord injury in rats. Three groups of male rats (n = 10 per group) were injured by compression of the cord at T1 for 1 minute with a 52-g clip. The hydrogen clearance technique was used to measure SCBF at the T1 segment. Motor and somatosensory evoked potentials were recorded. SCBF and evoked potentials were measured before injury and again at approximately 1 and 2.5 hours after injury. The methylprednisolone group received a bolus of methylprednisolone (30 mg/kg) at 5 minutes after injury and then at 15 minutes after injury, the group received an infusion of methylprednisolone at 5.4 mg/kg per hour. The nimodipine group received placebo at 5 minutes and then received an infusion of nimodipine at 0.02 mg/kg per hour at 15 minutes. The placebo group received placebo at both times. Physiological parameters were closely monitored and maintained within the normal range. Albumin was administered after injury to maintain mean arterial blood pressure at or above 80 mm Hg. The infusions were continued for approximately 3 hours after spinal cord injury. SCBF was not significantly different between the experimental groups at either 1 or 2.5 hours postinjury (P = 0.16 and 0.71, respectively), and evoked potential responses did not return in any rat at any time after injury. Thus, this experiment failed to demonstrate an improvement in SCBF or electrophysiological function with either drug.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413880 TI - Dexamethasone decreases the delivery of tumor-specific monoclonal antibody to both intracerebral and subcutaneous tumor xenografts. AB - The effect of dexamethasone on the delivery of monoclonal antibody L6 IgG to intracerebral and subcutaneous LX-1 small cell lung carcinoma xenografts was evaluated in nude rats (n = 157). Dexamethasone (0, 8, or 24 mg/m2) was given 18 hours before infusion of L6 IgG, with or without osmotic disruption of the blood brain barrier. Compared with controls, the 8 mg/m2 dose decreased delivery of L6 IgG (12-37%) to all tissues, but the only significant decrease (P < 0.001) was in the subcutaneous tumor (37%). In the 24 mg/m2 group, L6 IgG delivery was significantly (P < 0.001) decreased to all tissues (37-60%). Dexamethasone had no effect on plasma levels. Barrier disruption significantly (P < 0.0001) increased L6 IgG delivery to intracranial tumor and surrounding brain, but not to subcutaneous tumor or plasma. The percentage of decremental effect of dexamethasone on L6 IgG delivery was the same with and without barrier disruption and was not associated with the time the animals were killed (P > 0.05). Compared with controls, the ratio of intracranial tumor to normal brain showed no change with dexamethasone, but the ratios of both intracranial and subcutaneous tumors to plasma significantly (P < 0.002) decreased with both doses. The in vitro cell binding capacity of L6 IgG to LX-1 cells remained unchanged after incubation of cells with dexamethasone over a 3-log concentration for 4 days, demonstrating no effect on antigen expression. This study suggests that dexamethasone has a clinically relevant generalized (i.e., central nervous system and systemic) vascular effect on permeability to L6 IgG monoclonal antibody. PMID- 8413881 TI - Determination of the lethal dose of dexamethasone for early passage in vitro human glioblastoma cell cultures. AB - Previous investigators have supported the idea that glucocorticoids may be oncolytic. In this study, the percentage of cell death in two human glioblastoma cell cultures was related to the concentration of dexamethasone that was administered. It was determined that for Cell line 1, the median lethal dose was approximately 500-800 micrograms/ml and the completely lethal dose was about 900 1000 micrograms/ml; the 3H-thymidine uptake to approximate the mitotic rate was 16,607 cpm, and the dexamethasone receptor activity was 228 fmol/mg protein. The median lethal dose and completely lethal dose for Cell line 2 was approximately 500-600 micrograms/ml and 700-1000 micrograms/ml, respectively; the 3H-thymidine uptake was 8402 cpm, and the dexamethasone receptor activity was 137 fmol/mg protein. These lethal concentrations of dexamethasone are probably higher than can be tolerated by systemic delivery. However, it remains to be seen whether the interstitial administration of dexamethasone could achieve local concentrations resulting in the oncolysis of malignant gliomas. The clinical significance of these findings will depend on the local tolerance of normal brain parenchyma to very high doses of dexamethasone. A review of some of the literature is included. PMID- 8413882 TI - Modulation of proliferation and antigen expression of a cloned human glioblastoma by interleukin-4 alone and in combination with tumor necrosis factor-alpha and/or interferon-gamma. AB - As part of continuing studies to investigate the possible regulatory effects of cytokines on malignant astrocytes, we investigated the effects of interleukin-4 (IL-4) alone and in combination with tumor necrosis factor-alpha (TNF alpha) and/or interferon-gamma (IFN gamma) on the cell growth and major histocompatibility complex (MHC) antigen expression of a cloned human glioblastoma cell line (9C). The 9C cells were treated with IL-4 alone or in combination with TNF alpha and/or IFN gamma and were examined for proliferation by crystal violet assay and for Class II MHC antigen by flow cytometry. Results indicated that IL-4 alone did not affect 9C proliferation. In combination with TNF alpha or IFN gamma, however, IL-4 significantly and dose-dependently inhibited cell growth. As previous reports have shown, TNF alpha combined with IFN gamma exerted an additive growth suppressive effect on glioblastoma cells, probably by enhancing TNF receptor expression. This additive effect of TNF alpha and IFN gamma was further enhanced by IL-4. In contrast, IL-4 did not modulate expression of Class II MHC antigen on 9C cells, even in combination with IFN gamma, which predictably enhanced this antigen. These results suggest that IL-4 is capable of modulating glioblastoma growth only in the presence of other cytokines, such as TNF alpha and/or IFN gamma. Further, the effect of IL-4 on glioblastoma proliferation is selective and independent of the mechanisms involved in regulating MHC antigen expression. PMID- 8413883 TI - Protein kinase C inhibitors suppress cell growth in established and low-passage glioma cell lines. A comparison between staurosporine and tamoxifen. AB - We have previously demonstrated that the proliferation of established human glioma cell lines correlated with protein kinase C (PKC) activity and that a relatively selective PKC inhibitor, staurosporine, inhibits glioma cell proliferation. The purpose of this study was to determine whether low-passage glioma cell lines were also sensitive to staurosporine and to compare the antimitotic effects of staurosporine with tamoxifen, an antiestrogen with a known PKC inhibitory effect presently being investigated in the treatment of recurrent glioma. We measured the effects of treatment with staurosporine or tamoxifen on the proliferation rate of five established glioma cell lines (A172, U251, U87, U373, U563) and four low-passage glioma cell lines. The proliferation of all cell lines was inhibited by staurosporine, at an IC50 value (concentration at which activity is 50% inhibited) of approximately 2 nmol/L. All established lines, but only one low-passage line, were susceptible to tamoxifen, with an IC50 value of 10 mumol/L. Three of the four low-passage lines were poorly inhibited by tamoxifen. The IC50 values for the inhibition of cellular proliferation by staurosporine and tamoxifen closely corresponds to the IC50 data for the inhibition of particulate PKC activity in gliomas. We conclude that staurosporine is more effective in the inhibition of glioma proliferation than tamoxifen and that staurosporine is potentially useful in the adjuvant treatment of gliomas. The correspondence in IC50 results for proliferation and PKC activity further strengthens the hypothesis that an aberrant PKC system in gliomas drives their hyperproliferative state. PMID- 8413884 TI - Preoperative embolization of brain and spinal hemangioblastomas. AB - Large hemangioblastomas can be difficult to resect because of excessive bleeding. We report our experience with two patients whose large hemangioblastomas were embolized preoperatively and were totally resected with minimal blood loss and satisfactory postoperative outcome. Embolizations were carried out within 3 days of surgery, with the patient under general anesthesia, with 150- to 250-microns Contour emboli. A 65-year-old woman with progressive, severe myelopathy from a 4.5 x 2.1 x 1.4 cm intramedullary hemangioblastoma at T4 underwent embolization of the left T2 and T3 intercostal arteries; a 29-year-old man with Lindau's syndrome and posterior fossa hemorrhage from a 3.8 x 1.8 x 1.8 cm right cerebellar hemangioblastoma underwent embolization of the right posterior inferior cerebellar artery, two right anterior inferior cerebellar arteries, and a dural branch of the right vertebral artery. Embolization led to the complete obliteration of the tumoral blush without neurological sequelae. Both lesions were completely resected with a blood loss of 50 to 100 ml, with bleeding occurring mainly from the pseudocapsule of the lesion. Preoperative embolization of hemangioblastomas is a useful and relatively safe procedure that reduces blood loss at the time of surgery and allows complete resection. PMID- 8413885 TI - Primary pineal melanoma: case report. AB - Primary melanomas of the central nervous system are unusual, and those in the pineal region are exceedingly rare. We present a case of primary pineal melanoma in a 60-year-old man. The lesion was subtotally resected through an infratentorial, supracerebellar approach. The clinical features and the histological findings are discussed. Eight previous case reports are reviewed. PMID- 8413886 TI - Metastasis of malignant struma ovarii to the cranial vault during pregnancy. AB - Malignant struma ovarii is a rare type of ovarian teratoma; only 16 cases with distant metastases have been reported previously. We report an extremely rare case of malignant struma ovarii metastatic to the cranial vault, which developed during pregnancy. A 28-year-old woman in the 26th week of pregnancy, who had undergone resection of an ovarian tumor 3 years previously, noticed a mass in her left frontal region that had enlarged gradually in 6 months. Magnetic resonance imaging revealed a massive extradural tumor growing through the cranium. Under fetal heart monitoring, the patient underwent total resection of the tumor, including the adjacent cranial bone and dura, and a healthy infant was delivered at full term. Pathological examination showed that the tumor was a follicular adenocarcinoma. Because there was no abnormality in the patient's thyroid gland, this tumor was considered to be a metastasis from the ovarian tumor, a malignant struma ovarii, resected 3 years previously. The management of brain tumor during pregnancy is also discussed. PMID- 8413887 TI - Amyloid destructive spondyloarthropathy causing cord compression: related to chronic renal failure and dialysis. AB - Destructive spondyloarthropathy is a recently recognized disease that has not been reported in the neurosurgical literature. It is associated with spinal amyloid deposition in long-term renal failure and dialysis, and it occurs increasingly as the number of dialysis patients and their survival times increase. Clinically, there is a multisegmental and often rapidly progressive radiculomyelopathy that may require emergency stabilization. The radiological features are disc space narrowing with erosion of vertebral end plates and subarticular cysts. The pathological features include deposition of amyloid, which stains with Congo Red and antibodies to beta-2-microglobulin. We present two cases with clinical, radiological, and pathological features and a review of the literature. PMID- 8413888 TI - Intraoperative protection of cranial nerves and arteries by split silicone tube. AB - The authors describe the usefulness of split silicone tubing to protect the cranial nerves and arteries during microneurosurgery. The inner diameter of the tube varied from 1.0 to 3.3 mm with a thickness of 0.125 mm. Application of the tube protects the nerves and arteries from mechanical trauma, electrical injury, and dryness. PMID- 8413889 TI - Interchangeable and adjustable-length microneurosurgical instruments. AB - A new system of microneurosurgical instruments is presented in which a number of shafts with different tips are interchangeable and the length of the shafts are adjusted easily. With this system, the surgeon's tasks in microneurosurgery are quicker and smoother. PMID- 8413890 TI - Surgical management of high jugular bulb in acoustic neurinoma via retrosigmoid approach. PMID- 8413891 TI - Systemic approach to spinal reconstruction after anterior decompression for neoplastic disease of the thoracic and lumbar spine. PMID- 8413892 TI - Endoscopic cure of a giant sacral meningocele associated with Marfan's syndrome: case report. PMID- 8413893 TI - Surgical treatment of syringomyelia based on magnetic resonance imaging criteria. PMID- 8413894 TI - Targeted brain biopsy: a comparison of freehand computed tomography-guided and stereotactic techniques. PMID- 8413895 TI - Comment; ceftizoxime versus vancomycin and gentamicin in neurosurgical prophylaxis. PMID- 8413896 TI - A model for in utero endoscopic treatment of myelomeningocele. AB - An animal model for intrauterine surgical treatment of myelomeningocele is described using sheep. We report the technical feasibility of endoscopic intrauterine skin graft placement over surgically induced defects, including over exposed spinal cord. These grafts exclude amniotic fluid from the underlying lesion and provide a matrix for fetal skin growth below the graft. The potential for human application is discussed. PMID- 8413897 TI - Cross-modal associations and the human amygdala. AB - The role of the human amygdala in cross-modal associations was investigated in two subjects: SM-046, who had bilateral damage circumscribed to the amygdala; and the patient known as Boswell, whose damage in both temporal lobes includes the amygdala and surrounding cortices. Neither subject was impaired on Tactile-Visual or Visual-Tactile cross-modal tasks using the Arc-Circle test, suggesting that the amygdala is not involved in cross-modal associations involving perceptually "equivalent" basic stimulus properties. On the other hand, the results are compatible with the amygdala's involvement in higher-order associations between exteroceptive sensory data and interoceptive data concerned with correlated somatic states. PMID- 8413898 TI - Encoding ability is preserved in amnesia: evidence from a direct test of encoding. AB - The encoding ability of 17 amnesics of mixed aetiology and 17 matched normal controls was assessed directly using a novel procedure. On two separate occasions, subjects were shown 60 complex drawings each containing six pictures. On one occasion each drawing was shown for 6 sec, and on the other occasion it was shown for 25 sec. Immediately after presentation of each drawing subjects were asked a single unpredictable question about picture colours, location, size or semantic category. Amnesics performed normally in the 6-sec exposure condition indicating that all of the tested kinds of information were encoded at a normal rate. Performance in this condition correlated with short-term, but not long-term memory in the amnesics indicating that it depended largely on encoding and short term memory. However, the amnesics were impaired in the 25-sec condition where performance should have depended on (long-term) memory abilities at which they were impaired. The results are inconsistent with available encoding deficit accounts of amnesia. PMID- 8413899 TI - The role of right side objects in left side neglect: a dissociation between perceptual and directional motor neglect. AB - Based on a test introduced by Tegner and Levander, Brain 114, 1943-1951, 1991, right brain-damaged patients were assigned to a group with unilateral perceptual neglect and a group with directional motor neglect. Brain scans showed that all directional motor neglect patients had frontal lesions, whereas in perceptual neglect patients the frontal lobes were always spared. All patients were asked to execute two tasks, which were also administered to a control group. One task consisted in pointing to tokens symmetrically distributed on a display. The other task consisted in picking up the same tokens. The tasks were first executed with the aid of vision and then in a blindfolded condition. In the case of patients with perceptual neglect, performance on the left side was better in the pick-up task than in the pointing task and improved in the blindfolded condition. Neither patients with directional motor neglect nor control patients showed these effects. The results were explained in terms of the hyperattentional hypothesis of perceptual neglect, according to which, in this form of neglect, attention is captured by the objects that lie on the right side of space. PMID- 8413900 TI - Recognition and naming of famous faces in Alzheimer's disease: a cognitive analysis. AB - A Famous Faces Test designed to assess face recognition, spontaneous naming, verbal identification of un-named faces, and cued naming using semantic and phonetic cues was administered to 22 patients with dementia of Alzheimer type (DAT) and 25 matched controls. The DAT patients were significantly impaired in all test conditions with evidence of a temporal gradient for recognition, identification and naming with phonemic cues, but not for spontaneous naming or naming with semantic cues. Analysis of the effects of disease severity showed that performance in all five conditions of the test declined with increasing disease severity, but this did not reach significance for recognition or for naming with phonemic cues. The DAT patients identified and named a significantly smaller proportion of the faces that they recognized than did the controls, and at no stage was identification significantly better than spontaneous naming. These findings indicate that the primary deficit was not one of name access, but an actual loss of stored knowledge about the person represented. In keeping with this observation, semantic cueing did not aid naming. These findings are discussed in the context of contemporary cognitive models of face processing. PMID- 8413901 TI - Preserved pattern completion priming for novel, abstract geometric shapes in amnesics of several aetiologies. AB - Two experiments were conducted using a paradigm developed by Gabrieli et al., Neuropsychologia 28, 417-427, 1990, which assessed both indirect and direct memory performance in a completion task for novel abstract geometric patterns. The preferred method of scoring was the lines method, based on the number of correct and incorrect lines produced for each item. It was chosen because it is both the simplest and the most informative measure. Two methods of scoring were used in previous work, namely, the strict whole figure method and the lenient whole figure method (Gabrielli et al., 1990; Verfaelie et al., Brain Cognit. 18, 34-45, 1992). Therefore to facilitate comparisons between studies and to determine the characteristics of different scoring methods, results with all three measures were included. In Experiment 1, two different encoding strategies of naming and copying were used in order to explore the relationship between indirect and direct memory performance. Indirect memory performance in the naming condition was at baseline whereas in the copying condition it was significantly above baseline. Cued recall did not differ across encoding conditions but recognition was higher in the naming condition than the copying condition. In Experiment 2, an attempt was made to extend the findings of two studies, one with H.M. (Gabrielli et al., 1990) and one with nine Korsakoffs (Verfaelie et al., 1992), to a larger group of 14 amnesics of several aetiologies. Indirect memory performance was found to be equivalent for the amnesics and their matched controls, only when the lenient and the lines methods of scoring were used. Recognition and cued recall performance was impaired for the amnesics compared to the controls. PMID- 8413902 TI - Temporal lobe surface area measurements on MRI in normal and dyslexic readers. AB - In a neuroanatomical study of dyslexia, measurements were made of the superior surface of the temporal lobe (SSTL) on MRI scans in a sample of 17 dyslexics and 21 non-dyslexic subjects. Both anterior and posterior halves of the SSTL area showed significant leftward asymmetry in non-dyslexics, but showed symmetry in dyslexics. The total SSTL area showed greater leftward asymmetry in non-dyslexics than in dyslexics. The dyslexics also revealed a significant correlation (r = 0.69, P = 0.005) between Woodcock-Johnson Passage Comprehension scores and posterior SSTL asymmetry, such that those with higher scores had more leftward asymmetry. This suggests that among dyslexics the direction of SSTL asymmetry may serve as a risk factor and/or a marker for the severity of reading comprehension problems. PMID- 8413903 TI - Autobiographical recollection and frontal damage. AB - A neuropsychological experiment on autobiographical retrieval of incidental past events, checked by means of a standardized enquiry, was carried out on a series of 16 patients with CT-assessed frontal lobe lesions. Of 16 patients, six were impaired on autobiographical retrieval. Moreover, eight were impaired on supraspan verbal learning. Impairment on both resulted in three patients, whereas in five there was no impairment on either test. Concordance between autobiographical and learning impairment was far from significant. Poor autobiographical retrieval correlated significantly with "executive" test performances and with the CT-verified bilaterality of the frontal damage. Poor learning of new information did not correlate significantly with "executive" performances, nor did it appear to be related to the hemispheric side or to the bilaterality of the frontal lesion. We propose that the relationship between the attentional system of retrieving from remote memory and of learning new information is qualitatively different. PMID- 8413905 TI - Brain potentials in a phonological matching task using Chinese characters. AB - In readers of English, involved in a rhyme judgement task, mismatch trials are associated with an enhanced N450 component of the Event Related Potentials (ERPs). It has been suggested that N450 is related to orthographic or phonological priming. In this paper ERPs were recorded during a phonological matching task, using pairs of logographically dissimilar Chinese characters. A pair was considered to match if they sounded alike with identical phoneme sequences. The subjects (native Chinese speakers) were instructed to ignore vowel inflections, which in Chinese have lexical status. Since sublexical assembly of phonology is not used in reading Chinese characters, and the members of each pair were logographically dissimilar, match and mismatch trials did not suffer in the amount of orthographic or sublexical phonological priming. An enhanced negative component (latency near 400 msec), was observed in ERPs elicited by the second character in non-matching pairs. The negativity could be similar to N450. If this were so, then N450 could not be associated with orthographic priming, nor with sublexical phonology, but would probably be associated with postlexical processing. Also, in both readers of Chinese and English, the negativity enhanced in non-match trials is larger over the right side of the scalp, suggesting a similar brain lateralization of the underlying processes. PMID- 8413904 TI - Electrophysiological correlates of hierarchical stimulus processing: dissociation between onset and later stages of global and local target processing. AB - In the discussion of whether the processing of hierarchically structured stimuli proceeds from the more global to the more local level or vice versa, it is frequently assumed that the relative speed of global/local target identification (response time (RT) advantage) and the direction of interference from local/global distractors reflect the order of processing. Studies both in brain injured patients and in normals, however, have demonstrated that RT advantage and interference are dissociable, leading to the conclusion that they do not provide a valid index of the order of global/local processing. The aim of the present event-related brain potential (ERP) study was to assess electrophysiological correlates of global/local processing and to determine how the relative speed of responding to global and local targets is related to these ERP measurements. In a divided-attention paradigm, subjects were asked to respond to hierarchically structured letter stimuli that contained a target letter either at the global or at the local level. The behavioral results confirmed a dissociation between RT advantage and interference. ERP analysis revealed an early posterior negative component (denoted as N250) as a sign of early global/local target perception. It was found that RT advantage is not a valid measure of the onset nor of the time course of this component. Furthermore, the N250 components to global and local targets exhibited a different time course and a different topographical distribution, suggesting that they are determined by separate processing structures. Together, the behavioral and electrophysiological results support the view that global and local target perception may be mediated by separate brain systems acting, at least initially, in parallel. PMID- 8413906 TI - Hemispheric asymmetry for the auditory recognition of true and false statements. AB - A group of right-handed adult subjects (24 males and 24 females) were asked to judge phrases as to their truthfulness of falsehood, by listening to them via earphones either with the right or with the left ear. These phrases, which had been previously recorded by two non-actors (1 male and 1 female) and two actors (1 male and 1 female), were either true or false. The number of correct judgements made by the subjects was significantly higher than the number of wrong judgements. The subjects correctly recognised significantly more true phrases than false ones. Moreover, they better recognised true phrases pronounced by the actors and false phrases pronounced by the non-actors. True phrases pronounced by male speakers and false phrases pronounced by female speakers were better recognised. A left-ear advantage was found in the recognition of true statements. PMID- 8413907 TI - The behavioral effects of the destruction of the magnocellular basal nucleus of the forebrain of cats. AB - Behavioral experiments were carried out in cats following methodology which simulates complexly organized, nonautomatized behavior with elements of generalization and abstraction. A conclusion was reached regarding the participation of this formation in the structural-functional support of complex integrative forms of activity, cognitive and gnostic processes, was reached on the basis of the results of the performance of test tasks by the animals with partial destruction of the magnocellular basal nucleus. The proposed mechanism of the involvement of the basal nucleus in gnostic and cognitive processes is the nonspecific support of the system of structures which participate directly in thinking and learning. PMID- 8413908 TI - A spectral correlation analysis of the electrical activity of the sensorimotor cortex and the medial geniculate body in the presence of a motor polarizational dominant. AB - The dynamics of the changes in the structure of the coherence relationships of the electrical activity of the sensorimotor cortex and the medial geniculate body (MGB) of the rabbit in the presence of a motor polarizational dominant created by the action of a direct current anode on the region of the sensorimotor cortex were investigated by the method of spectral correlation analysis. It was demonstrated that, in addition to an increase in the components of the delta range in the interstimulus intervals in the presence of a dominant, a clear maximum in the alpha rhythm range in the electrical activity of the MGB of the "dominant" hemisphere appears in the power spectra of the electrical activity of the MGB during the action of an acoustic signal. An increase in the coherence of the delta range of the electrical activity of the MGB and of the sensorimotor cortex in the "dominant" half of the brain is manifested exclusively in the period of action of the acoustic stimulus. PMID- 8413909 TI - The influence of stimulation of the hypothalamus on the choice of food during maintenance of rats on various salt and water diets. AB - Electrical stimulation of the lateral hypothalamus of rats maintained on a normal or salty diet with water deprivation leads to an increase in drinking excitability and to a decrease in the thresholds for salty food consumed. The stimulation promotes attenuation of the effects of the "mental" desalting of food that is achieved by a spatial co-placement of the nonsaltiness signal with salty reinforcement, and accelerates the conversion of the signal significance of the conditional stimuli associated with nonsalty and salty reinforcement as well. Electrical stimulation of the hypothalamus accelerates the formation of conditioned reflex aversion to salty food and the inhibition of conditioned reflexes in the direction of the corresponding food dispenser, and intensifies the existing state of thirst as well, imparting to it characteristics of a dominant motivation. PMID- 8413910 TI - The dynamic of thermal fields of the brain of rats in the late post-resuscitation period under stress. AB - The temperature topography of the cerebral cortex of rats following clinical death and resuscitation was investigated with the technique of thermoencephaloscopy. Complete restoration of the neurological status of the animals was achieved over the course of one to two days. Marked disturbances in the background thermal maps and the thermal reactions of the cerebral hemispheres induced by a stressor were identified in the remote post-resuscitation period (up to two months). A pathological mosaicism of the thermal characteristics was detected under the conditions of relative rest, as were disturbances in the dynamics of the temperature reactions of the brain and in the character of the interhemispheric asymmetries under stress. The individual character of the post resuscitation pathology which is found both in the baseline thermal maps as well as following a functional load is emphasized. The results of the investigation point to the importance of an individual approach in the rehabilitation therapy of the post-resuscitation illness. PMID- 8413911 TI - The activity of Krebs cycle enzymes in the visual analyzer of rats in the norm and under stress. AB - Higher activity of the NAD-dependent dehydrogenases of the tricarboxylic acid cycle (TAC) is observed in the optic retina, and of FAD-dependent dehydrogenases in the occipital lobes of the brain, in the visual analyzer of intact rats. The influence of stress using Desiderato's method induces a compensatory increase in the activity of succinate dehydrogenase. Acute stress induces a change in the regulation of the activity of the TAC dehydrogenases, assessed on the basis of the reaction to functional load. The animals' remaining in the dark following stress promotes the restoration of the activity of the TAC cycle to the normal level. PMID- 8413912 TI - The influence on the realization of alimentary conditioned reflexes of the chronic injection of leu-enkephalin and its analog tetrapeptide into the caudate nucleus of dogs. PMID- 8413913 TI - The influence of microinjections of serotonin and dopamine into the dorsal raphe nucleus on the extinction of a conditioned reflex in rats. PMID- 8413914 TI - Destruction of the catecholaminergic system in newborn rats impedes the formation of cholinergic innervation of the neocortex. PMID- 8413915 TI - A comparative analysis of learning and exploratory behavior of rats with varying resistance to stressor influences and the level of brain monoamines. AB - Experiments have demonstrated that Wistar rats resistant (R) to acoustic stimulation differed from nonresistant (NR) rats by elevated exploratory activity under the condition of moderate stress in an open field test, by heightened reactivity to sensory stimuli of varying modality (somatosensory, visual, and olfactory), and by a reduced level of exploratory behavior in a burrow chamber. The rats of the R group differ from the NR animals by a greater capacity to learn a goal-directed reaction and by a lower capacity for discrimination of emotionally distinct influences. The results of a biochemical analysis of the content of biogenic amines in particular brain structures revealed in the resistant rats an increase in the level of norepinephrine, and in the non resistant rats, a higher level of dopamine and serotonin. PMID- 8413916 TI - The neuropharmacological restoration of cognitive functions of cats following a lesion to the basal nuclei of the forebrain (Meynert's nucleus). AB - The functions of generalization and abstraction in an experimental model of Alzheimer's disease (the lesion of Meynert's nuclei in cats) were studied against the background of stimulation and inhibition of the cholinergic, GABAergic, and dopaminergic systems of the brain. It was demonstrated that the cholinergic system is a key system for the formation of the function of generalization, the dopaminergic system improves simple forms of learning, while the GABAergic system is actively involved in the establishment of complex types of associations. PMID- 8413917 TI - Morphofunctional changes in the thyroid gland in different variants of chronic experimental stress. PMID- 8413918 TI - The features of the influence of psychotropic agents on the structural-functional state of the adrenals of intact and stressed animals. PMID- 8413919 TI - Investigation of neurons of the cell populations of the nuclei tractus solitarius in rats of different sexes. PMID- 8413920 TI - Localization of aspartate aminotransferase in structures of a human sensory neuron. PMID- 8413921 TI - The effect of increased nitrogen pressure on motor activity and the intercentral relationships of the brain of monkeys. AB - The influence of increased nitrogen pressure up to 19 ata was investigated in chronic experiments on monkeys of a species of Javan macaques (Macaca irus) with electrodes implanted in cortical and subcortical regions of the brain. The phasic character of the changes of motor activity of the animals, an increase in the spectral density of the average power of the EEG in the range of frequencies from 4 to 20 Hz, and a disturbance in the connectedness of the electrogenesis of the reticular formation of the midbrain with the bioelectrical processes in the substantia nigra, the head of the caudate nucleus, and the frontal and motoric areas of the cortex were identified in the course of nitrogen compression at a rate of 1.0 ata per 1 min. PMID- 8413922 TI - The ultrastructural bases of the effect of haloperidol on the brain. PMID- 8413923 TI - Antisense oligodeoxynucleotide inhibits D2 dopamine receptor-mediated behavior and D2 messenger RNA. AB - There are several subtypes of dopamine receptors in the central nervous system which mediate the actions of dopamine in producing its diverse motor and behavioral effects. In this study we determined whether an antisense oligodeoxynucleotide directed to the mRNA encoding one of the subtypes of the dopamine receptor can inhibit a specific dopamine-mediated behavior. Accordingly, the effects of a phosphorothioate-modified antisense oligodeoxynucleotide targeted toward the D2 dopamine receptor mRNA (D2 antisense) was studied in mice with unilateral 6-hydroxydopamine-induced lesions of the corpus striatum. Rotational behavior in response to different agents, and the levels of D2 and D1 dopamine receptors and D2 and D1 dopamine receptor mRNAs in corpus striatum were then measured. In control mice, lesioning resulted in a contralateral rotational behavior in response to the D1 dopamine receptor agonist SKF 38393, the D2 dopamine agonist quinpirole, and the muscarinic cholinergic agonist oxotremorine. Lesioning also caused an increase in D2 dopamine receptor mRNA levels in the dorsolateral striatum. Intraventricular injections of the D2 antisense inhibited rotational behavior induced by quinpirole but not that induced by SKF 38393 or that induced by oxotremorine. Repeated administration of the D2 antisense significantly reduced the levels of the D2 dopamine receptor and D2 dopamine receptor mRNA in the dorsolateral but not the dorsomedial striatum. Similar treatment failed to significantly alter the levels of the D1 dopamine receptor or D1 receptor mRNA in dorsolateral or dorsomedial striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413924 TI - CDRK and DRK1 K+ channels have contrasting localizations in sensory systems. AB - Molecular cloning of mammalian potassium channels has revealed an extensively heterogeneous superfamily of potassium channels derived from four basic subfamilies, Shaker, Shaw, Shal and Shab, each with multiple members. The families were first identified in Drosophila, in which subfamily heterogeneity is derived by alternative splicing, while in mammals mainly distinct genes give rise to channel subtypes. Further diversity of mammalian potassium channels is demonstrated by the identification of some which do not belong to any of the four main subfamilies. Although potassium channels are differentiated into fast inactivating and delayed rectifier types, differential functions of the many mammalian potassium channels are unclear. Moreover, potassium channels function as homotetramers, though in principle heterotetramers might have a physiological role as is the case with heteromers of neurotransmitter receptor subunits. Insight into differential functions of potassium channels may be provided by their regional and subcellular localizations. In the rat brain in situ hybridization and immunohistochemistry have revealed distinct regional localizations for various subfamilies. In one instance a particular subfamily predominated in cell bodies and another in axons. We demonstrated dramatically different localizations for two members of the Shab subfamily, circumvallate papilla delayed rectifier K+ channel (CDRK) and delayed rectifier potassium channel 1 (DRK1), which in major portions of their sequences display more than 90% amino acid identity. In a number of brain regions they occur in distinct neuronal cell types or subcellular compartments, with CDRK predominantly localized diffusely over soma and in fibers and DRK1 most evident in soma and dendritic process. PMID- 8413925 TI - Heat shock protects neuronal cells from programmed cell death by apoptosis. AB - The programmed cell death (apoptosis) of a proportion of the neurons which form plays a critical role in the development of the nervous system and ensures that the correct number of mature neurons are ultimately present. We show that the prior exposure of neuronal cells to an elevated temperature sufficient to induce the heat-shock response partially protects the cells from apoptotic cell death following subsequent transfer to serum-free medium. The degree of protection observed in experiments using different heat-shock or recovery times correlates with the extent of heat-shock protein synthesis. Similarly activation of heat shock protein synthesis by inducers other than elevated temperature also results in protection from apoptosis. The mechanism by which the heat-shock proteins may protect neuronal cells from apoptosis is discussed. PMID- 8413926 TI - Protective effects of SR 57746A in central and peripheral models of neurodegenerative disorders in rodents and primates. AB - Compounds possessing neurotrophic properties may represent a possible treatment for neurodegenerative disorders such as Alzheimer's disease. SR 57746A, 1-[2 (naphth-2-yl)ethyl]-4-(3-trifluoromethylphenyl)-1,2,5,6- tetrahydropyridine hydrochloride, is a new compound with neurotrophic activity in a number of in vitro preparations. The neurotrophic effects of this compound have been evaluated in vivo using four distinct rat models of neurodegeneration: transient global ischaemia produced by a four-vessel occlusion; septohippocampal lesion produced by injection of vincristine sulphate into the medial septum; sciatic nerve crushing; and acrylamide-induced peripheral neuropathy. Rats were administered vehicle or 2.5-10 mg/kg p.o. SR 57746A, after initiation of the degenerative process, then once daily for 10 days in the first two models, 16 days in the third and 26 days in the fourth model. Median scores for ischaemia-induced neuronal damage were reduced by 30-40% by SR 57746A treatment in hippocampal CA1, CA2, and CA3 regions, and in the dorsal striatum. Twelve days after intraseptal vincristine administration, there was a marked loss of septohippocampal cholinergic neurons, as indicated by reduced choline acetyltransferase activity in both the septum and hippocampus. SR 57746A dose-dependently reversed this reduction in both areas. These results were confirmed by histoenzymological evaluation of hippocampal acetylcholinesterase content. SR 57746A also reversed the loss of hippocampal choline acetyltransferase induced by intraseptal vincristine in marmosets. Behavioral deficits in these models (exploratory behaviour in the former and short-term social memory in the latter) were also significantly reduced by SR 57746A treatment. In the sciatic crush model, sensorimotor function improved more rapidly in rats treated with 10 mg/kg SR 57746A. In this same model, SR 57746A (10 mg/kg/day) also significantly increased the length of regenerated nerve eight days after the crush, as measured using the pinch test. Finally, SR 57746A retarded the onset, reduced the amplitude and accelerated the recovery of acrylamide-induced peripheral neuropathy. Thus, SR 57746A possesses notable neurotrophic activity in a variety of neurodegenerative models in vivo, suggesting that the compound may possess therapeutic potential for the treatment of neurodegenerative diseases. PMID- 8413927 TI - Block of long-term potentiation by intracellular application of anti phosphatidylinositol 4,5-bisphosphate antibody in hippocampal pyramidal neurons. AB - We examined the effects of black widow spider toxin and anti-phosphatidylinositol 4,5-bisphosphate antibody on the changes in excitatory postsynaptic currents and spontaneous excitatory postsynaptic currents accompanying long-term potentiation using whole-cell recording from hippocampal CA1 pyramidal neurons of rodents. In the presence of black widow spider toxin, tetanic stimulation of input fibers produced a short-lived potentiation followed by a gradual decline of the excitatory postsynaptic current amplitude. With an anti-phosphatidylinositol 4,5 bisphosphate antibody containing pipette, tetanus elicited only decremental potentiation of excitatory postsynaptic currents with a reduced frequency of spontaneous excitatory postsynaptic currents, suggesting inhibition of retrograde reinforcement from the antibody-injected neuron. With both black widow spider toxin and anti-phosphatidylinositol 4,5-bisphosphate antibody, neurons showed a rapid depression of excitatory postsynaptic currents after tetanus. The results indicate that time-dependent interactions between presynaptic terminals and the postsynaptic spikes take place during long-term potentiation. PMID- 8413928 TI - Changing the light intensity of the visual environment results in large differences in numbers of synapses and in photoreceptor size in the retina of the young adult rat. AB - A quantitative light- and electron-microscopic study has been made of the retinae of rats which were exposed to different lighting conditions for between one and 15 weeks in young adulthood, having been reared in identical conditions during development. The width of the inner and outer segments of the photoreceptors and the width of the outer plexiform layer varied inversely with the light intensity under diurnal lighting conditions of 10 h light/14 h dark. Linear regression analysis showed that the widths were inversely related to the fourth root of the light intensity as measured in lux. Both central and peripheral areas of retina showed a similar change. No change was seen in the widths of the inner plexiform layer, or of the inner and outer nuclear cell layers. Nor was there a difference in the packing density or size of the nuclei in the nuclear cell layers. The number of ribbon synapses in the outer plexiform layer also varied inversely with the intensity of diurnal light. Linear regression analysis showed that the number of synapses was inversely correlated with the fourth root of the light intensity and was positively correlated with the width of the outer plexiform layer. The number of ribbon synapses was increased by up to two and a half times in constant darkness compared to diurnal light of 35 lux. The increase was present but not maximal after one week of exposure. The length of synaptic ribbons was unchanged. The nerve terminals forming such synapses were increased in size but not in number. After one week, there was little or no additional change in the retinal widths and number of synaptic ribbons with time. However, there was a progressive increase with time in nerve terminal size (two-fold in area) in constant darkness. There was some evidence of a slight decrease in nerve terminal number and increase in size of retinal nuclei with age. It is concluded that the adult retina responds to a different lighting environment by a relatively rapid change in the size of photoreceptor segments, by a progressive and large change in number of ribbon synapses and by a slower progressive and large change in the size of photoreceptor nerve terminals. The response is quantitatively determined by the strength of the stimulus but not in a linear fashion. These results are compared with the effects of environmental stimulation of other areas of the nervous system. PMID- 8413929 TI - Immunocytochemistry and in situ hybridization of neuropeptide Y in the hypothalamus of Xenopus laevis in relation to background adaptation. AB - The amphibian Xenopus laevis is able to adapt to a dark background by releasing melanophore-stimulating hormone from the pars intermedia of the pituitary gland. The inhibition of melanophore-stimulating hormone release is accomplished by neuropeptide Y-containing axons innervating the pars intermedia. To determine the production site of neuropeptide Y involved in this inhibitory control, the distribution of neuropeptide Y in the brain has been investigated by immunocytochemistry and in situ hybridization. Immunoreactive cell bodies were visualized in, among others, the ventromedial and posterior thalamic nuclei, and the suprachiasmatic and ventral infundibular hypothalamic nuclei. A positive hybridization signal with a Xenopus-specific probe for preproneuropeptide Y-RNA was found in the diencephalic ventromedial thalamic nucleus and in the suprachiasmatic nucleus. With both immunocytochemistry and in situ hybridization, suprachiasmatic neurons appeared to be stained only in animals adapted to a white background; animals adapted to a black background showed no staining. Quantitative image analysis revealed that this effect of background adaptation is specific for suprachiasmatic neurons because no effect could be demonstrated of the background light condition on the ventral infundibular nucleus (immunocytochemistry) or the ventromedial thalamic nucleus (in situ hybridization). These results indicate that neurons in the suprachiasmatic nucleus enable the adaptation of X. laevis to a white background, by producing and releasing neuropeptide Y that inhibits the release of melanophore-stimulating hormone from the melanotrope cells in the pars intermedia of the pituitary gland. PMID- 8413930 TI - Effect of rapid eye movement sleep deprivation on rat brain monoamine oxidases. AB - Monoamine oxidase, monoamine oxidase-A, and monoamine oxidase-B activities were compared in free moving, rapid eye movement sleep-deprived, recovered, and control rat brains. The activities were estimated in the whole brain, cerebrum, cerebellum, whole brainstem, medulla, pons, and midbrain. The flowerpot method was used for continuing deprivation for one, two, or four days. Monoamine oxidase activity decreased significantly in the cerebrum and the cerebellum of the sleep deprived rats, whereas monoamine oxidase-A and monoamine oxidase-B were differentially affected. Medullary MAO-A was the first to be affected, showing an increase after just one day of rapid eye movement sleep deprivation, while longer deprivation decreased its activity. The activity of monoamine oxidase-B was not significantly affected in any brain areas of the deprived rats until after two days of rapid eye movement sleep deprivation. All the altered enzyme activities returned to control levels after recovery. Control experiments suggest that the decrease was primarily caused by the rapid eye movement sleep deprivation and was not due to nonspecific effects. These findings are consistent with past studies and may help to explain earlier observations. The results support the involvement of aminergic mechanisms in rapid eye movement sleep. The plausible reasons for the changes in the activities of monoamine oxidases, after rapid eye movement sleep deprivation, are discussed. PMID- 8413931 TI - Simultaneous intracellular recordings from enteric neurons reveal that myenteric AH neurons transmit via slow excitatory postsynaptic potentials. AB - Simultaneous intracellular electrical recordings were made from pairs of neurons separated circumferentially by 100-200 microns of the myenteric plexus of the guinea-pig ileum in vitro. The recording electrodes were filled with the dye neurobiotin which was injected into impaled nerve cells, and later revealed histochemically. Intracellular current pulses were used to evoke action potentials via the recording electrode in one type of myenteric neuron, in most cases an AH neuron, while a second electrode was used to record from a simultaneously impaled S neuron or AH neuron. AH neurons are thought to be primary sensory neurons, whereas S neurons are interneurons and motor neurons. Ninety pairs of neurons were adequately tested for interaction. From these, 17 S neurons and three AH neurons that responded to AH neuron stimulation were detected. In each case, the response was a slow depolarization that was seen only in response to a train of stimuli at 10 Hz. The slow depolarizations were enhanced by passing depolarizing current and diminished by hyperpolarization. Responses were also diminished by lowering external Ca.2+ and elevating Mg2+. In all cases in which intracellular recording indicated communication between neurons, morphological evidence of connection was seen. In no case was there communication without connection, but in four instances, morphological connections appeared to exist, although no physiological evidence of communication was obtained. PMID- 8413932 TI - Corticotropin-releasing factor release from the mediobasal hypothalamus of the rat as measured by microdialysis. AB - Procedures were developed to permit the measurement of corticotropin-releasing factor in perfusate collected from microdialysis probes implanted in various brain areas of anesthetized and awake rats. Initially in vitro experiments were carried out to optimize the recovery of corticotropin-releasing factor and the radioimmunoassay conditions. Addition of a specific antiserum against corticotropin-releasing factor to the perfusion medium (artificial cerebrospinal fluid) increased the relative in vitro recovery over a range of different flow rates (1-10 microliters/min) using commercially available microdialysis probes with a membrane cutoff of 20,000 mol. wt. This procedure increased recovery from 3% to 6% at flow rate of 2.5 microliters/min, and from 4% to 8% at a flow rate of 5 microliters/min. In vivo experiments were performed with a flow rate of 3.3 microliters/min and 50-microliters fractions were used for radioimmunoassay. In each experiment, the standard curve of the radioimmunoassay was constructed from aliquots of the same medium used to perfuse the probe. Basal levels of corticotropin-releasing factor in dialysate collected from the mediobasal hypothalamus of anesthetized rats were estimated to be 0.75 +/- 0.07 fmol/50 microliters. Raising the concentration of potassium (60 mM) in the perfusate increased corticotropin-releasing factor levels to 2.04 +/- 0.37 fmol/50 microliters. Hypertonic stress induced by intraperitoneal injection of 1.5M NaCl (20 ml/kg) elevated the levels to 1.32 +/- 0.07 fmol/50 microliters. A marked increase of corticotropin-releasing factor levels was also produced by a 10-min pulse of the potassium-channel blocker 4-aminopyridine (10 mM) included in the perfusate. A second stimulation pulse with 4-aminopyridine, administered 2 h after the first pulse again increased the levels, with a mean ratio between the first and second pulse of 0.97. Corticotropin-releasing factor efflux produced by the second stimulation pulse was completely inhibited by perfusion with calcium free medium containing calcium-chelating agent ethyleneglycol tetraacetic acid (10 mM). In separate experiments, microdialysis probes were implanted in several brain areas of anesthetized rats. Basal and potassium-evoked levels of corticotropin-releasing factor were measured in dialysate collected from the amygdala (1.20 +/- 0.22 and 2.05 +/- 0.48 fmol/50 microliters, respectively) and frontal cortex (0.51 +/- 0.10 and 1.64 +/- 0.15 fmol/50 microliters, respectively). Corticotropin-releasing factor levels in the dorsal part of the third ventricle and in the striatum were below the detection limits. In awake rats, corticotropin-releasing factor levels in the mediobasal hypothalamus were 0.98 +/- 0.03 fmol/50 microliters.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8413933 TI - Differential regulation of melanin-concentrating hormone gene expression in distinct hypothalamic areas under osmotic stimulation in rat. AB - Melanin-concentrating hormone and associated peptides represent a novel peptide neuronal system that may be involved in the control of water homeostasis in mammals. We have examined the effect of 24 h dehydration or salt-loading over a period of six days, on melanin-concentrating hormone messenger RNA levels in rat brains by using complementary methods of Northern blotting and in situ hybridization histochemistry. In response to one to six day salt-loading regimen, hypothalamic melanin-concentrating hormone messenger RNA content in male or female rats decreased by two to three-fold. Levels of melanin-concentrating hormone messenger RNA in the hypothalamus were also dramatically decreased following dehydration in female rats whereas contrasting responses were noted in male rats. In addition, no significant variation in the low levels of melanin concentrating hormone gene transcripts in medulla pons and cortex was found after osmotic stimulus. In agreement with Northern blot data, in situ hybridization studies revealed that the majority of the melanin-concentrating hormone expressing neurons in the anterior part of the lateral hypothalamus of dehydrated or salt-loaded rats expressed lower amounts of melanin-concentrating hormone messenger RNAs than those found in control rats. Interestingly, less variation was found in the posterior part of the lateral hypothalamus. Furthermore few clusters of cells, located in zona incerta and near the internal capsula and fornix, increased their contents in melanin-concentrating hormone messenger RNA in salt-loaded but not in dehydrated rats suggesting that melanin-concentrating hormone gene expression may be regulated differently by various osmotic stimuli. Finally, diurnal variations in melanin-concentrating hormone messenger RNA contents were observed in normal and dehydrated rats with highest levels around 22.00 h and lowest levels during daylight hours. However, the up-regulation of melanin-concentrating hormone gene activity at night was found lower in dehydrated rats than in control animals suggesting that osmotic stress may interfere with the generation of the diurnal pattern of melanin-concentrating hormone messenger RNA expression. Altogether, our results indicate that osmotic stimulations lead to a selective and conspicuous inhibition of melanin concentrating hormone gene activity in the whole hypothalamus of rat. We suggest that the melanin-concentrating hormone neuronal system plays an important role in integration processes relative to nocturnal regulation of water homeostasis and drinking behavior. PMID- 8413934 TI - Glucocorticoid modulation of calcium currents in growth hormone 3 cells. AB - Clonal malignant pituitary growth hormone 3 cells were used for the analysis of the influence of hydrocortisone and dexamethasone on voltage-gated calcium currents and hormone secretion. The whole-cell patch-clamp technique was used. The presence of low-threshold inactivating and high-threshold persisting components in the total calcium current was shown; they could be separated at less negative holding potential level. Some increase in current densities of both components was observed as early as 30 min after treatment with 10(-6) mol/l glucocorticoids. The increase was maximal for both types of currents after 2 h of incubation; however, the high-threshold component was affected much more strongly (current density increased by more than four-fold) than the low-threshold one (current density increased by about a three-fold). Potentiation of currents was blocked by actinomycine D (10(-4) M), suggesting that protein synthesis was required. A substantial increase in growth hormone secretion (measured by radioimmunoassay method) was observed in the same cells after 2 h of incubation with hydrocortisone, while the secretion of prolactin remained even slightly depressed. PMID- 8413935 TI - Spinal and hindbrain structures involved in visceroception and visceronociception as revealed by the expression of Fos, Jun and Krox-24 proteins. AB - We have used the evoked expression of the immediate early gene-encoded proteins (Krox-24, c-Fos, Fos B, Jun D, Jun B, c-Jun) to monitor visceral processing in both the spinal cord and hindbrain structures of rats undergoing either mechanical colorectal or chemical intraperitoneal stimulation. Experiments were conducted under controlled volatile anaesthesia to suppress affective reactions that visceral stimulations may induce. The results refer to the effects of anaesthesia alone, and of both innocuous and noxious stimulations. Non nociceptive and nociceptive stimulation but not anaesthesia were effective in evoking c-Fos, c-Jun, Jun B and Krox-24 expressions in the spinal cord. Intraperitoneal injections labelled cells mostly at the thoracolumbar junction levels, while colorectal distension labelled cells mostly at the lumbrosacral junction levels. Labelling was widely distributed throughout the gray matter including superficial layers, deep dorsal horn, lamina X and sacral parasympathetic columns. Krox-24- and, to a lesser degree, c-Jun-labelled cells were quite numerous in the superficial layers of the dorsal horn; Jun B, and especially c-Fos, were very effective in demonstrating inputs to all parts of the spinal cord. Both anaesthesia and noxious visceral stimulation were effective in evoking c-Fos, Krox-24 and Jun B expressions in discrete hindbrain subregions. The structures which are primarily labelled under anaesthesia are the rostral ventrolateral medulla, the external medial and lateral nuclei of the parabrachial area, the medial and dorsal subnuclei of the nucleus of the solitary tract, the area postrema, the central gray including pars alpha and nucleus O, the nucleus beta of the inferior olive, the locus coeruleus, and the inferior colliculi and adjacent parts of central gray. The structures which are primarily labelled following noxious visceral stimulation are the caudal intermediate reticular nucleus as part of the caudalmost ventrolateral medulla and the superior lateral nucleus of the rostrolateral parabrachial area. Labelling in the caudal intermediate reticular nucleus was maximal for colorectal distension. Labelling in the superior lateral nucleus was specific to peritoneal inflammation. The Edinger-Westphal nucleus is a structure in which noxious-evoked labelling was superposed onto the anaesthesia-evoked labelling. Nociception-evoked overexpression in this nucleus was maximal for intraperitoneal inflammation. The present work demonstrates that the central effects induced by either anaesthesia or visceroception including pain can be effectively monitored through the induction of an array of immediate early genes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8413936 TI - Calretinin immunoreactivity in the monkey hippocampal formation--I. Light and electron microscopic characteristics and co-localization with other calcium binding proteins. AB - Calretinin-containing neurons were visualized by immunocytochemistry in the monkey hippocampal formation, subicular complex, and entorhinal cortex. Calretinin-immunoreactivity was present exclusively in non-granule cells of the dentate gyrus and in non-pyramidal cells of Ammon's horn, subiculum and entorhinal cortex. Most frequently, calretinin-positive neurons were found at the hilar border of the dentate granule cell layer and in the stratum radiatum of CA1 3 areas. In the subicular complex, immunoreactive neurons were evenly distributed in all layers, whereas in the entorhinal cortex, they were accumulated in external layers above the lamina dissecans. Distinct bands of calretinin-positive fibers occupied the supragranular zone of the molecular layer in dentate gyrus, the pyramidal cell layer of the CA2 area in Ammon's horn and the upper two layers of presubiculum. The majority of calretinin-immunoreactive neurons were small, bipolar or fusiform neurons with a dendritic tree oriented parallel to the dendrites of principal cells (granule cells in dentate gyrus and pyramidal neurons elsewhere). Dendrites were smooth or sparsely spiny, displaying small spines of conventional type. Co-existence studies showed that these neurons were completely devoid of other calcium-binding proteins, parvalbumin and calbindin. Electron microscopic analysis revealed somata of immunoreactive neurons which contained a large nucleus and a small cytoplasmic rim, which contained only few organelles. The nucleus displayed deep infoldings and intranuclear rods. Input synapses of immunoreactive neurons were rare both on somata and dendrites and large surface areas were frequently apposed by glial processes. This was very prominent in the dentate gyrus and Ammon's horn. Axons of calretinin-positive neurons were thin, arborized in all layers and had small varicosities. Their terminals formed symmetric synaptic contacts mainly with dendrites and less frequently with somata of principal cells. Axon terminals of calretinin immunoreactive fiber bundles in the supragranular layer, as well as in the pyramidal layer of the CA2 area, formed asymmetric synaptic contacts with dendritic shafts. In addition, they established asymmetric axospinous and axosomatic synaptic contacts with granule cells of the dentate gyrus. In the presubiculum, the calretinin-positive axon bundle included a large number of immunoreactive myelinated axons, as well as axon terminals. The characteristic location and features of synapses suggests that these fibers derive from extra hippocampal afferents (Nitsch, R. and Leranth C. (1993) Neuroscience 55, 797-812) and not from the calretinin-immunoreactive neurons of the hippocampal formation. PMID- 8413937 TI - Massive exocytosis triggered by sodium-calcium exchange in sympathetic neurons is attenuated by co-culture with cardiac cells. AB - Entry of Ca2+ through voltage-dependent Ca2+ channels is known to be linked to the exocytotic release of transmitter from sympathetic neurons. In this paper we provide evidence that transmitter release can also be stimulated by Ca2+ influx via the Na-Ca exchanger. Furthermore, the release linked to Na-Ca exchange is regulated by cardiac target cells. Cultured sympathetic neurons of the chick embryo incubated in Ca2(+)-Mg(2+)-free Krebs solution for 20 min and then switched to Ca(2+)-containing solution exhibited 15-20-fold increases in [3H]noradrenaline release over the spontaneous release. Electrophysiologic studies showed that neurons were completely depolarized in Ca(2+)-Mg(2+)-free medium. Indo-1 fluorescence revealed a large and sustained increase in intracellular free Ca2+ concentration ([Ca2+]i) after addition of Ca2+ to Ca(2+) Mg(2+)-free medium. The increased [3H]noradrenaline release and [Ca2+]i were dependent on external Na+ and Ca2+, but were not affected by the Ca2+ channel blockers lanthanum, cadmium, verapamil or omega-conotoxin. A conventional depolarizing stimulus (125 mM K+) produced a 13-fold increase in [3H]noradrenaline release over spontaneous release. However, K(+)-induced release and rise in [Ca2+]i declined rapidly and were sensitive to the Ca2+ channel blockers. When sympathetic neurons were co-cultured with embryonic cardiac cells the release induced by change from Ca(2+)-Mg(2+)-free to Ca(2+)-Krebs solution was dramatically reduced. The change from Ca(2+)-Mg(2+)-free to Ca(2+)-Krebs solution was ineffective in evoking [3H]noradrenaline release from sympathetic neurons in situ using perfused hearts of 15-day-old chick embryos.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413938 TI - Ontogeny of 5-hydroxytryptamine2 receptor immunoreactivity in the developing rat brain. AB - In this study, we investigated the regional and temporal emergence of 5 hydroxytryptamine2 receptor immunoreactivity in the developing rat brain. In a qualitative immunocytochemical analysis using an antibody against the rat 5 hydroxytryptamine2 receptor protein, we visualized cells expressing the receptor in the pontine tegmentum, caudate nucleus, basal forebrain, hippocampus and neocortex of developing rats. Three potentially important periods in the developmental regulation of 5-hydroxytryptamine2 receptors were identified: the time of onset, a period of accelerated expression and hyper-elaboration, and a period of regression. In general, the onset of 5-hydroxytryptamine2 receptor immunoreactivity occurred relatively late in the ontogeny of cells in these regions, in the late prenatal and early postnatal periods. Following the perinatal onset of receptor expression, there was a rapid increase in the number of immunoreactive neurons during the first week after birth. In neocortex, there appeared to be a relative over-expression of the receptor, with an elevated density and hyper-elaboration of immunopositive neurons relative to the adult, reaching a peak at the end of the second week. There was then a gradual decrease in both the density and morphological complexity of cortical 5-hydroxytryptamine2 labelled neurons, until the adult pattern of expression was achieved at about four weeks of age. In all areas studied, cells positive for the 5 hydroxytryptamine2 receptor were first detected within the regions in which they would ultimately reside, and after the known periods of cell proliferation for these regions. These observations would argue against a role for the 5 hydroxytryptamine2 receptor as a transducer of the early developmental influences of serotonin in the central nervous system, but leave open the possibility that the receptor may participate in regulating some aspect of terminal differentiation or late maturation of the neurons on which it is found. The identification of important developmental periods in the ontogeny of 5 hydroxytryptamine2 receptors suggests time-points at which events that disrupt the normal ontogenetic pattern of expression could produce long-lasting effects on central serotonergic neurotransmission. PMID- 8413939 TI - Physical trauma and multiple sclerosis. PMID- 8413940 TI - The use of animals in science. PMID- 8413941 TI - Trauma and multiple sclerosis: a population-based cohort study from Olmsted County, Minnesota. AB - Utilizing the Olmsted County, Minnesota, population-based records-linkage resource at Mayo Clinic, we identified an incidence and a prevalence cohort with multiple sclerosis (MS), a head injury cohort, and a lumbar disk surgery cohort to evaluate the association between mechanical trauma and MS onset or exacerbation. The MS cohorts consisted of 225 incidence cases (1905 to 1991) and 164 prevalence cases (December 1, 1991) of definite MS in the population of Olmsted County. We assessed the effect of mechanical trauma in the form of spinal injury or extremity fracture with regard to precipitation of MS or exacerbation of an existing neurologic deficit. Fifty-four episodes of trauma, as defined, occurred among 39 MS prevalence cases; most occurred 10 years or more after the onset of disease and were associated with existing MS-related disability. We compared the final disability status of the groups with and without trauma. We found no correlation between the occurrence of peripheral fractures and the onset of MS, exacerbation of MS, or final disability due to MS in the prevalence cohort. In a cohort of 819 head injury cases from the Olmsted County population, none developed MS within 6 months of the trauma. In a lumber disk surgery cohort of 942 local residents, there were five with MS, but onset of MS had preceded the spinal surgery in four of the five. Thus, we found no association of head injury and spinal disk surgery with onset of MS. PMID- 8413942 TI - Neurologic function in the optimally healthy oldest old. Neuropsychological evaluation. AB - We examined cognition on a wide range of standardized neuropsychological tests in two groups of optimally healthy, elderly volunteers. One was composed of community-dwelling, functionally independent individuals aged 84 years and older, and the other group was nearly 20 years younger. The effect of aging was greatest on visual perceptual and constructional tasks rather than on memory tasks. Many cognitive functions were relatively well preserved in the optimally healthy oldest old. PMID- 8413943 TI - Is the incidence of dementing illness changing? A 25-year time trend study in Rochester, Minnesota (1960-1984) AB - We performed a time trend study of incidence of dementing illness in Rochester, Minnesota. We ascertained age- and sex-specific incidence rates for the five quinquennia, 1960 through 1984. The incidence rates sharply increased with advancing age, reaching a high of 2,922/100,000 person years in the group 85 years and older. For dementia caused either solely or predominantly by Alzheimer's disease, this figure was 2,600/100,000 person years for the oldest age group. There were no significant differences in the incidence of dementing illness between men and women. In the oldest age groups in the last two quinquennia of study, there appears to be a trend toward increasing incidence rates. Over the years, the proportion of cases attributed to dementia due to unknown causes has decreased while the proportion of cases attributed to Alzheimer's disease has increased. PMID- 8413944 TI - Auditory function in Alzheimer's disease. AB - The pattern of cerebral degeneration in Alzheimer's disease (AD) patients suggests that basic auditory capacities should be normal in AD, whereas progressively higher levels of auditory function should be increasingly impaired. To test this hypothesis, we administered tests of auditory capacities associated with primary auditory cortex (sound localization and perception of complex tones) and auditory association cortex (phoneme discrimination, timbre discrimination, and tonal memory) to 19 mildly to moderately demented AD patients, 21 elderly control subjects (ECS), and 14 young control subjects (YCS). The results showed significant differences between YCS and ECS on phoneme discrimination with synthetic speech and on tonal memory. The AD group differed from the ECS group on sound localization, one measure of synthetic speech discrimination, and timbre discrimination. Performance did not correlate with age, dementia severity, or duration of illness on any test condition. These findings indicate that although AD is accompanied by specific auditory deficits, the increase in neuropathologic change between primary auditory and auditory association cortices is not reflected in an increased impairment of functions that are mediated by these areas. Degraded aural language comprehension, which is characteristic of AD, likely reflects disruption of language processes, rather than dysfunction specific to auditory circuits. PMID- 8413945 TI - Nonoptic aphasia: aphasia with preserved confrontation naming in Alzheimer's disease. AB - Patients with Alzheimer's disease (AD) frequently exhibit anomia early in the course of the disease. Current theories of naming describe lexical retrieval in visual confrontation naming as requiring processing through the semantic system before lexical access. We report on three patients with AD who demonstrated severely impoverished spontaneous speech, impaired list generation, but preserved visual confrontation naming. Extensive investigation of one of these patients revealed an impairment of semantic knowledge. Our results support previous theories that there are two routes to visual naming, one via the semantic system and one directly from the internal visual representation to the lexicon. We labeled this aphasia resulting from an impaired semantic system but a preserved direct route to the lexical system as "nonoptic aphasia" and suggest that it is the earliest language deficit in some patients with AD. PMID- 8413946 TI - A relationship between particular reproductive endocrine disorders and the laterality of epileptiform discharges in women with epilepsy. AB - Cerebral limbic structures modulate the endocrine function of the hypothalamopituitary axis in both animals and humans. In an investigation of 30 women who had complex partial seizures with unilateral temporal lobe epileptiform discharges and reproductive endocrine disorders, there was a significant difference between the EEG laterality distributions associated with polycystic ovarian syndrome (PCO) and those associated with hypogonadotropic hypogonadism (HH). There was a strong predominance of left-sided discharges (15 versus one) with PCO, and right-sided discharges (12 versus two) with HH. Each distribution differed significantly from that of 30 women with epilepsy who had no reproductive endocrine disorder (left/right: 17/13). Moreover, among women with PCO who had unilateral non-temporal lobe foci, six of seven had right-sided epileptiform discharges. This represents a significant difference from the EEG laterality distribution among women with PCO who had temporal foci. These relationships between altered patterns of reproductive hormonal secretion and the predominant laterality of EEG epileptiform discharges in women with epilepsy are consistent with a lateralized asymmetry in cerebral influences on reproductive endocrine function. PMID- 8413947 TI - Serum anti-GQ1b IgG antibody is associated with ophthalmoplegia in Miller Fisher syndrome and Guillain-Barre syndrome: clinical and immunohistochemical studies. AB - To determine the significance of serum anti-GQ1b IgG antibody, we studied the disease spectrum associated with this antibody and GQ1b epitope in the human nervous system. We examined sera from 19 patients with typical Miller Fisher syndrome (MFS), five patients with acute postinfectious ophthalmoplegia without ataxia (atypical MFS), six patients with Guillain-Barre syndrome (GBS) with ophthalmoplegia (GBS-OP[+]), and 23 patients with GBS without ophthalmoplegia (GBS-OP[-]). We also examined sera from 84 patients with other neurologic or non neurologic disorders and from 16 normal control subjects. Eighteen of the 19 patients with typical MFS, all the patients with atypical MFS, and five of the six patients with GBS-OP(+) had increased anti-GQ1b IgG activity in ELISA, but none of the patients in the other groups, including GBS-OP(-), had it. All the patients' sera that had anti-GQ1b IgG antibody showed anti-GT1a IgG activity. Results of absorption studies suggested that the same antibody reacted with GQ1b and GT1a. An anti-GQ1b mouse monoclonal antibody immunostained the paranodal regions of the extramedullary portion of the human oculomotor, trochlear, and abducens nerves. Biochemical analysis showed that the human oculomotor nerve contained a larger amount of GQ1b than did the ventral and dorsal roots of the spinal cord. We conclude that serum IgG antibody against GQ1b is very closely associated with acute postinfectious ophthalmoplegia in MFS and GBS. PMID- 8413948 TI - Levodopa therapy and survival in idiopathic Parkinson's disease: Olmsted County project. AB - We studied survival in all Olmsted County Parkinson's disease (PD) patients seen at the Mayo Clinic from 1964 to 1978, attempting to answer two questions: (1) What effect does levodopa have on survival in PD? and (2) Does the timing of levodopa administration influence survival? We chose this period because it allowed us to study patient records with a spectrum of disease durations before levodopa treatment; in many patients, the treatment delay was exclusively due to levodopa being unavailable prior to 1969. Mortality of the entire PD cohort (N = 179; 61% levodopa-treated) was greater than that of the general population (matched chronologically, geographically, and by age and gender). Lower age at onset of motor symptoms, lower Hoehn and Yahr stage at first neurologic visit for parkinsonism, and treatment with levodopa were all independent predictors of improved survival. Using a time-dependent Cox regression model, we assessed the impact of the timing of levodopa administration during the course of illness on mortality, while statistically controlling for other factors (ie, patient selection for levodopa treatment, and independent predictors of survival). Risk of death following initiation of levodopa was significantly reduced (p < 0.001), regardless of pre-levodopa duration of illness. This reduction gradually diminished over a period of 4 years on levodopa, but continued to be significantly reduced. After 4 years, increasing survival benefit again progressively accrued over time to at least 17 years of levodopa treatment (p < 0.001). At no point in time was levodopa treatment associated with increased mortality, arguing against substantial levodopa toxicity. However, despite levodopa-improved survival, mortality continues to be increased in PD relative to the general population. PMID- 8413949 TI - Ocular motor nerve palsies in spontaneous dissections of the cervical internal carotid artery. AB - Cranial nerve palsies affecting ocular motor function in patients with spontaneous cervical internal carotid artery dissections are rare. Among 155 patients with spontaneous dissections of the cervical internal carotid artery, four (2.6%) had transient third, fourth, or sixth cranial nerve palsy. The third nerve was involved in two patients, the fourth nerve in one, and the sixth nerve in one. Three patients had ipsilateral headache or facial pain, one had bilateral headaches, and three had oculosympathetic palsy. None had any associated cerebral or retinal ischemic symptoms. Cervical internal carotid artery dissection should be included in the differential diagnosis of palsies of the third, fourth, or sixth cranial nerve, especially when associated with ipsilateral headache or facial pain. Interruption of the nutrient arteries supplying these cranial nerves could explain their involvement by internal carotid artery dissection. PMID- 8413950 TI - Sensory syndromes in parietal stroke. AB - We studied 20 patients with an acute parietal stroke with hemisensory disturbances but no visual field deficit and no or only slight motor weakness, without thalamic involvement on CT or MRI and found three main sensory syndromes. (1) The pseudothalamic sensory syndrome consists of a faciobrachiocrural impairment of elementary sensation (touch, pain, temperature, vibration). All patients have an inferior-anterior parietal stroke involving the parietal operculum, posterior insula, and, in all but one patient, underlying white matter. (2) The cortical sensory syndrome consists of an isolated loss of discriminative sensation (stereognosis, graphesthesia, position sense) involving one or two parts of the body. These patients show a superior-posterior parietal stroke. (3) The atypical sensory syndrome consists of a sensory loss involving all modalities of sensation in a partial distribution. Parietal lesions of different topography are responsible for this clinical picture, which probably represents a minor variant of the two previous sensory syndromes. Neuropsychological dysfunction was present in 17 patients. The only constant association was between conduction aphasia and right-sided pseudothalamic sensory deficit. We conclude that parietal stroke can cause different sensory syndromes depending on the topography of the underlying lesion. Sensory deficits can be monosymptomatic but never present as a "pure sensory stroke" involving face, arm, leg, and trunk together. PMID- 8413951 TI - Nonepileptic seizures and childhood sexual and physical abuse. AB - Nonepileptic seizures (NES) must be distinguished from epilepsy to avoid the adverse effects of unnecessary antiepileptic drugs and to initiate appropriate psychiatric treatment. A higher frequency of prior sexual abuse has been suspected in NES, although no prospective controlled study has compared patients with NES and epilepsy. A series of patients with conversion disorder presenting as epilepsy and 140 patients with complex partial epilepsy (CPE) without evidence of conversion were selected from a series of consecutive admissions to a comprehensive epilepsy center. The groups did not differ with respect to age, years of education, race, or marital status, but the percentage of women was greater in the conversion NES group (73.2%) than in the CPE control group (50.7%; p < 0.002). The frequency of a history of sexual or physical abuse was greater in the NES group (32.4%) than in the CPE controls (8.6%; p < 0.000). Severity of sexual but not physical abuse was significantly greater in the NES group relative to controls (p < 0.05). There was a trend for a closer relationship of the perpetrator of sexual abuse to the victim among the NES patients compared with CPE controls (p < 0.1). These results support the impression that childhood abuse is more common among patients with conversion NES than with epilepsy, and suggests that in some cases childhood abuse may be a contributory pathogenetic factor. PMID- 8413952 TI - Validity of family history data on severe headache and migraine. AB - This study evaluated the validity of family history data on severe headache and migraine obtained from adult probands with epilepsy in a family study of seizure disorders. We compared the probands' reports of severe headaches and migraines in their parents and full siblings with symptom-based diagnoses based on self reports. Overall sensitivity was 48% for severe headache and 44% for migraine. When we used reports of severe headaches to screen for migraine headaches in the relatives, sensitivity was 56%. These results indicate that severe headache and migraine are underreported in family history interviews with first-degree relatives. To obtain accurate information on headaches in families for clinical practice and genetic research, direct interviews with each relative may be required. PMID- 8413953 TI - Effects of radiation therapy on adult brain behavior: evidence for a rebound phenomenon in a phase 1 trial. AB - Although radiotherapy (XRT) is a necessary course of treatment to prolong life expectancy in patients with many types of brain neoplasms, it has damaging effects that are little understood. We used a comprehensive neuropsychological battery to evaluate five patients with low-grade brain tumors prior to XRT and then at 3-month intervals up to 9 months postcompletion of XRT. We matched patients by age and education with six normal control subjects to assess baseline impairment. In intrasubject comparisons, we examined change over time postcompletion of XRT. In spite of varying locations of tumors, all patients showed deterioration in long-term memory at a mean of 1.5 months postcompletion of XRT. Patients also consistently demonstrated a rebound effect between means of 4.7 and 7.6 months post-XRT. We found no changes over time in working memory, attention, visuospatial processes, or on any other neuropsychological test with the exception of information-processing speed, which quickened over time. Functional measures of fatigue and mood did not correlate significantly with the long-term memory scores. Long-term memory appears sensitive to the proposed white matter changes thought to be the mechanism for the early-delayed effects of XRT. Our findings suggest a neurobehavioral model for studying the effects of XRT on brain functioning. PMID- 8413954 TI - Frontal lobe epilepsy: clinical seizure characteristics and localization with ictal 99mTc-HMPAO SPECT. AB - We evaluated ictal 99mtechnetium hexamethyl propylene-amine-oxime single-photon emission computed tomography (SPECT) in 22 children with electroclinical features of frontal lobe epilepsy (FLE). Ictal SPECT demonstrated unilateral frontal hyperperfusion in 20 of 22 children (91%) (one lobar, two frontocentral, six dorsolateral, six frontopolar, three orbitofrontal, one medial frontal, and one insula), concordant with electroclinical lateralization in 19 of 20 (95%). Hyperperfusion was evident in the ipsilateral basal ganglia in 16 of 22 (73%) and the contralateral cerebellum in 14 of 22 children (64%). Interictal SPECT showed unilateral, localized frontal hypoperfusion concordant with electroclinical lateralization in only two of 22 children (9%). Ictal SPECT localization to the frontocentral, media frontal, or dorsolateral regions was associated with asymmetric tonic posturing, contralateral head/eye deviation, and unilateral clonic jerking (p < 0.01). Ictal SPECT localization to the frontopolar or orbitofrontal regions was associated with vocalization, hyperventilation, truncal flexion, and complex gestural automatisms (p > or = 0.05). Ictal SPECT has the potential to (1) localize seizures in patients with intractable FLE, and (2) advance understanding of the in vivo anatomico-clinical relationships of frontal lobe seizures. PMID- 8413955 TI - Monoamine oxidase B (MAO B) inhibitor therapy in Parkinson's disease: the degree and reversibility of human brain MAO B inhibition by Ro 19 6327. AB - The possibility of slowing the progression of Parkinson's disease (PD) with inhibitors of monoamine oxidase B (MAO B) has stimulated the development of new MAO B inhibitor drugs. Ro 19 6327 is a highly selective inhibitor of MAO B currently under clinical investigation. We used positron emission tomography (PET) and the MAO B tracer [11C]L-deprenyl to determine the degree and reversibility of human brain MAO B inhibition by Ro 19 6327 in six early Parkinson's disease patients who were treated with different doses of Ro 19 6327 (25 mg [n = 3], 50 mg [n = 2], and 100 mg [n = 1]; 0.34 to 1.4 mg/kg) every 12 hours for 1 week. Each patient had three PET scans to assess baseline MAO B activity, degree of trough inhibition, and reversibility. A control group of four elderly normal subjects was scanned twice to assess reproducibility of repeated measures. Four of the patients showed reduction of MAO B concentration to 1% to 7% of baseline on doses of 0.43 mg/kg or greater, and the remaining two at 0.34 mg/kg showed significant but incomplete inhibition (10% to 21% of baseline in the global region and in the thalamus, basal ganglia, and mesencephalon). Thus, 0.4 mg/kg or greater of Ro 19 6327 given every 12 hours is the minimum dose necessary to provide > 90% inhibition of brain MAO B in patients with early PD. Brain MAO B activity returned to baseline values by 36 hours after drug discontinuation. PMID- 8413956 TI - Isolated horizontal supranuclear gaze palsy as a marker of severe systemic involvement in Gaucher's disease. AB - Type 3 neuronopathic Gaucher's disease (GD3) is phenotypically heterogeneous. In many GD3 patients, progressive myoclonus and dementia dominate the illness, with death secondary to progressive CNS disease. We have designated this group as GD3a. We studied 14 children with Gaucher's disease, isolated horizontal supranuclear gaze palsy, and aggressive systemic disease, and designated this group as GD3b. In comparison with 13 children with type 1 non-neuronopathic Gaucher's disease, the GD3b children presented earlier, and were shorter, underweight, and more prone to cardiopulmonary, hepatic, and skeletal complications. One-half of the children died in childhood or adolescence of systemic complications. Patients with at least one copy of the mutation that causes substitution of asparagine for serine at amino acid 370 of glucocerebrosidase did not develop neurologic signs. Patients homoallelic for the mutation causing substitution of leucine for proline at position 444 had severe systemic disease; neurologic signs were frequently, but not invariably, present. Early diagnosis and timely enzyme replacement therapy promise to improve the prognosis in GD3b. PMID- 8413957 TI - In vivo cerebral metabolism and central benzodiazepine-receptor binding in temporal lobe epilepsy. AB - Positron emission tomography measured interictal cerebral glucose metabolism with [18F]fluorodeoxyglucose and central benzodiazepine-receptor binding with [11C]flumazenil in 10 mesial temporal lobe epilepsy (TLE) patients and in normal subjects. Eight TLE patients had mesial temporal, lateral temporal, and thalamic hypometabolism ipsilateral to EEG ictal onsets, with additional extratemporal hypometabolism in four. One had unilateral anterior mesial temporal hypometabolism only, and one had normal metabolism. Each patient had decreased benzodiazepine-receptor binding in the ipsilateral anterior mesial temporal region, without neocortical changes. Thus, interictal metabolic dysfunction is variable and usually extensive in TLE, whereas decreased central benzodiazepine receptor density is more restricted to mesial temporal areas. Metabolic patterns in TLE may reflect diaschisis, while benzodiazepine-receptor changes may reflect localized neuronal and synaptic loss that is specific to the epileptogenic zone. [11C]Flumazenil imaging may be useful in presurgical evaluation of refractory complex partial seizures. PMID- 8413958 TI - Measurement of whole temporal lobe and hippocampus for MR volumetry: normative data. AB - We measured the volumes of the entire length of temporal lobe and hippocampal formation from coronal images in 29 healthy young adults to take into account normal side-to-side variation. Although whole-brain, temporal lobe, and left hippocampal volumes were significantly smaller in women, normalizing measurements for the whole brain eliminated intersex temporal lobe and hippocampal differences. There was a weak but significant inverse correlation between age and normalized hippocampal, but not temporal lobe or whole-brain, volume. In contrast to previous studies, we found no significant side-to-side differences in the sizes of temporal lobes or hippocampi. When performing MR volumetry, it is important to include the entire length of the hippocampal formation. PMID- 8413959 TI - Pregnancy and delivery in Charcot-Marie-Tooth disease type 1. AB - We report the obstetric complications and the influence of pregnancy and delivery in 21 Charcot-Marie-Tooth disease type 1 (CMT 1) patients with 45 gestations. Sixteen patients had subjective disabilities from childhood or youth, and five with late onset had subclinical CMT when they became pregnant. The rate of obstetric complications in the study group was in accordance with that of the normal population, and there was no deleterious effect on fetal outcome. Of the 21 participants, 38% (8 of 21) reported an exacerbation of CMT in at least one pregnancy. These patients noted increasing weakness in 81% (17 of 21) of their gestations. A temporary worsening occurred in 35% (6 of 17) of these pregnancies, and neurologic disabilities persisted after 65% of the deliveries (11 of 17). Patients who had pregnancy-associated progression in the first gestation (7 of 21) experienced similar deterioration in subsequent pregnancies (10 of 11), ie, there is a high risk for recurrence of exacerbations. Four women (19%) stated that their last deliveries were responsible for either an exacerbation or the onset of the neuropathy. The remaining nine patients (43%) denied any effect of their gestations on the progression of the neuropathy. Among the patients who had subjective disabilities from childhood or youth, the risk of a noticeable exacerbation in at least one pregnancy was 50% (8 of 16) and affected 81% of their gestations (17 of 21), whereas there was no influence of pregnancy in the five patients with adult onset of CMT, although the first symptoms were noticed postpartum in two of them.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8413960 TI - Fatigue in Parkinson's disease. AB - Fatigue is a common but poorly understood symptom in Parkinson's disease (PD). Using previously validated scales, we asked 58 nondemented PD patients and 58 controls to fill out questionnaires assessing fatigue and depression and found that PD patients were more depressed and more fatigued than age-matched controls. Although fatigue correlated with depression but not with disease severity, many nondepressed patients had significant complaints of fatigue. PMID- 8413961 TI - Fulminant multiple system atrophy in a young adult presenting as motor neuron disease. AB - A 34-year-old man demonstrated rapidly progressive motor neuron disease and, late in his 9-month clinical course, exhibited ophthalmoplegia and dysautonomic symptoms. Neuropathology showed spinal and bulbar motor neuron disease with severe involvement of extraocular motor nuclei, degeneration of spinal sympathetic and bulbar parasympathetic nuclei, striatonigral degeneration, and early olivopontocerebellar atrophy. This case underscores the diversity of multiple system atrophy and demonstrates an unusually rapid course in a young patient. PMID- 8413962 TI - Choroid plexus infection in cerebral toxoplasmosis in AIDS patients. AB - We evaluated the postmortem incidence of choroid plexus infection in cerebral toxoplasmosis in 17 patients with acquired immune deficiency syndrome (AIDS) and cerebral toxoplasmosis and, by immunohistochemistry, identified Toxoplasma gondii tachyzoites in this structure in 53% of all cases. They were present in 78% of the nine cases with the acute necrotizing stages of CNS toxoplasmosis but were less frequent (20%) in patients with only the healed cystic lesions of toxoplasmosis. Large necrotizing abscesses of the choroid plexus were found in three of the patients. In one of these, the choroid plexus was the sole site of CNS infection, which presented as radiographically documented masses in the third and fourth ventricles associated with obstructive hydrocephalus. These results demonstrate that infection of the choroid plexus is common with cerebral toxoplasmosis and suggest that this infection should be included in the differential diagnosis of intra- or periventricular lesions in patients with AIDS. In addition, the high frequency of choroid plexus infection with acute cerebral toxoplasmosis suggests that cerebral toxoplasmosis in the immunosuppressed patient may be due to hematogenous spread to the choroid plexus from reactivation of latent organisms from systemic organs rather than to reactivation of latent organisms within the brain itself. Furthermore, the high frequency of choroid plexitis offers the potential for CSF dissemination of this infection. PMID- 8413963 TI - Genomic imprinting and anticipation in idiopathic torsion dystonia. AB - Idiopathic torsion dystonia (ITD) is a dominantly inherited disorder with variable penetrance and expressivity. Factors affecting the penetrance of the ITD gene have not yet been identified. The present study used four published series of cases to test specific hypotheses regarding factors that could affect the expression of ITD. Among the combined 253 families, transmission of ITD did not depend on either the sex of the affected offspring or that of the transmitting parent. Furthermore, neither the specific type of dystonia manifested, the site at which clinical signs of dystonia first appeared, nor age of onset differed significantly as a function of the gender of the transmitting parent. However, in familial cases of later onset (age > or = 20 years), nearly all involved a transmitting mother. There is evidence for consistency of age of onset within the subset of Jewish families. Although anticipation effects are apparent, sampling bias cannot be ruled out. PMID- 8413964 TI - A new peroxisomal disease with impaired phytanic and pipecolic acid oxidation. AB - Phytanic acid (PA) accumulates in patients with adult Refsum disease (ARD) and with peroxisomal disorders. In three related patients with ARD, PA levels were moderately increased in plasma, whereas phytanic oxidation was severely deficient in the fibroblasts. Two of these patients had a significant increase of pipecolic acid in plasma, a finding not reported in ARD, and a fourth related patient, a brother, died at age 17 from a progressive neurologic disorder with unusual clinical and neuropathologic (a spongy degeneration of the white matter) abnormalities for ARD. The first step of L-pipecolic acid degradation occurs in peroxisome. In these patients, the accumulation of PA could have resulted from an impaired capacity to degrade pristanic acid rather than PA. The activity of pristanic oxidase, measured in fibroblasts, was normal, as were two other peroxisomal enzymes, lignoceric acid oxidase and dihydroxyacetone phosphate transferase. Since both mitochondria and peroxisomes are involved in PA alpha oxidation, we propose that these four related patients presented various phenotypical variants of a novel peroxisomal disease with impairment of PA and pipecolic acid oxidation. PMID- 8413965 TI - Type IIa ('anti-Hu') antineuronal antibodies produce destruction of rat cerebellar granule neurons in vitro. AB - We reacted dispersed cultures of newborn rat cerebellar granule cells with serum, purified IgG, and CSF from patients with type IIa ("anti-Hu") antibody response accompanying paraneoplastic neurologic syndromes. All type IIa sera, IgGs, and CSFs, but not those of normal or cancer controls, produced bright nuclear immunofluorescence of cultured granule neurons. Type IIa serum and CSF labeled proteins of 35-42 kd in rat granule cell blots, identical in molecular weight to proteins labeled by type IIa antibodies in blots of human granule cells. IgGs eluted from the 35-42 kd band in blots of rat granule cells labeled proteins of similar molecular weights in blots of human granule cells and produced typical type IIa immunostaining of human cerebellar sections. Human IgG could be identified in nuclei and cytoplasm of neurons incubated for 72 hours with 2/4 type IIa sera tested, but not with normal sera. Type IIa sera or IgGs from 4/7 patients produced specific lysis of rat granule cells in the presence of complement, as compared with controls using normal serum or heat-inactivated complement. Prolonged (7-day) incubation of cultures with type IIa antibody without complement also resulted in specific lysis, whereas incubation with normal serum or serum from neurologically normal patients with small-cell carcinoma of the lung did not. Rat granule cell cultures provide a valuable in vitro system with which to study the interaction of type IIa antibody with neurons. The present study provides the first reported evidence that type IIa antibodies may cause cell injury directly, in the absence of lymphocyte-mediated immune response. PMID- 8413966 TI - Brain imaging in late-onset GM2 gangliosidosis. AB - We describe brain CT and MRI characteristics of 10 patients with late-onset GM2 gangliosidosis. Cerebellar atrophy, particularly of the vermis, was a prominent feature in all patients with normal-appearing cerebral hemispheres. The severity of these findings did not correlate with the age of onset, disease duration, severity of neurologic impairment, or mode and distribution of the various clinical presentations. In particular, no cerebral abnormality was found by neuroimaging in seven patients with intellectual decline and in six patients with recurrent psychosis, while prominent cerebellar atrophy was present in the only patient who was free of cerebellar signs. PMID- 8413967 TI - Correlation of evoked potential and MRI findings in Wilson's disease. AB - We recorded brainstem auditory evoked potentials (BAEPs) and somatosensory evoked potentials to median nerve stimulation (MSEPs) within 10 days of brain MRI in 20 patients with Wilson's disease (WD). MRI was abnormal in 90% of patients, demonstrating symmetric striatum and brainstem lesions with or without thalamic lesions. MSEPs were abnormal in 65% of patients, usually showing bilaterally prolonged N/P13-N20 latencies. BAEPs were abnormal in 40%, most often with bilateral prolongation of the III-V latency. The III-V and N/P13-N20 interpeak latencies correlated significantly with the severity of MRI lesions in the caudal pons, rostral pons, and caudal midbrain. Our results indicate that subclinical sensory dysfunction is common in WD, and that auditory and somatosensory pathways are most severely affected at the brainstem level. Both the localization and severity of evoked potential abnormalities correspond closely to the morphologic changes in the pons and caudal midbrain shown by MRI. PMID- 8413968 TI - Proton magnetic resonance spectroscopy in Creutzfeldt-Jakob disease. AB - We studied two patients with Creutzfeldt-Jakob disease by in vivo proton magnetic resonance spectroscopy and obtained spectra from an extract of biopsy tissue from a third patient. In vivo spectra from the two patients, 3 months and less than 1 month after symptom onset, revealed only minor changes. A second study of one of the patients 10 months after symptom onset found a decrease in N-acetylaspartate and other metabolites. Spectroscopy of the biopsy extract obtained 4 months after onset of symptoms showed no reduction in metabolites measured by in vivo spectroscopy, in accord with quantitative pathology showing no overall neuronal loss. Changes in metabolites detectable by proton magnetic resonance spectroscopy are not an early feature of this disease. PMID- 8413969 TI - Anticardiolipin antibodies are an independent risk factor for first ischemic stroke. The Antiphospholipid Antibodies in Stroke Study (APASS) Group. AB - We evaluated 255 consecutive first ischemic stroke patients and 255 age-/sex matched hospitalized nonstroke patients at 15 medical centers for anticardiolipin antibodies (aCL). Sera were obtained within 7 days of stroke onset (cases) or hospital admission (controls) and stored at -20 degrees C until the aCL level was determined at a central laboratory. Results were read as negative, low, moderate, or high positive based on standardized optical density values. We recorded clinical, laboratory, and radiologic data without knowledge of antibody status. A positive aCL level was present in 9.7% of stroke patients and 4.3% of controls. The odds ratio for stroke status, given aCL positivity, was 2.31 after adjustment for age, ethnicity, gender, hypertension, diabetes mellitus, coronary artery disease, and cigarette smoking. Thus the frequency of aCL is significantly increased in these patients with first ischemic stroke. Further, aCL appear to be an independent risk factor for stroke in these patients. PMID- 8413970 TI - The clinical spectrum of cerebral amyloid angiopathy: presentations without lobar hemorrhage. AB - Cerebral amyloid angiopathy is a common cause of spontaneous lobar hemorrhage in elderly patients. We discuss seven patients with amyloid angiopathy presenting without major lobar hemorrhage. The patients' presentations fell into two groups: recurrent transient neurologic symptoms and rapidly progressive dementia. The cases with transient episodes had a spread of symptoms to contiguous body areas during episodes. Each had evidence of small hemorrhage or subsequent large hemorrhage in the cortical location corresponding to the symptoms, suggesting petechial hemorrhage with focal seizure as the cause of the transient spells. Three cases of dementia developed with relatively rapid time courses, progressing from intact baseline to profound dementia in spans of a few days to 2 years. Pathologic abnormalities, in addition to amyloid angiopathy, included patchy white matter demyelination and tissue loss, petechial cortical hemorrhages, cortical infarctions, and a variable degree of neuritic plaques and neurofibrillary tangles. The clinical spectrum of cerebral amyloid angiopathy includes these two neurologic syndromes. PMID- 8413971 TI - Evidence for neuronal degeneration and dendritic plasticity in cortical pyramidal neurons of Huntington's disease: a quantitative Golgi study. AB - Aided by a computer microscope, the Eutectic Neuron Tracing System, we performed a quantitative analysis of 59 rapid Golgi-impregnated pyramidal neurons from the third and fifth prefrontal cortical layers (P III and P V neurons) in tissue sections obtained from seven autopsied Huntington's disease (HD) patients (grades 2 through 4) and 59 corresponding cells from eight age-matched control cases. Relative to controls, P III HD neurons had a significant increase in the number of primary dendritic segments arising from soma, total dendritic length, and total surface area. The HD cells also had significantly more dendritic branches at three intervals of measurement in a Sholl diagram (100 microns, 200 microns, and 400 microns from the soma) and a significant increase in the number of dendritic branching points. The dendritic spine density in P III HD neurons was comparable to that of control subjects and significantly lower than that in P V HD cells. The total number and the total density of dendritic swellings were significantly increased in both P III and P V neurons, being most numerous in grades 2 and 3 cases. Rare withered cells with shrunken dendritic trees, harboring few spines and numerous varicosities on their dendritic shafts, were present in HD but not in control cases. Thus, while a small fraction of prefrontal cortical pyramidals degenerates in HD, the plasticity of the remaining pyramidal neurons, evidenced as an orderly augmentation of the dendritic tree, may represent a compensatory response sufficient to maintain relatively normal metabolic function of the cortex in most adult-onset cases. PMID- 8413972 TI - Familial meningioma is not allelic to neurofibromatosis 2. AB - Meningiomas frequently lose parts of chromosome 22 (CHR 22), suggesting that a meningioma tumor-suppressor gene is located on CHR 22. Since meningiomas are common in neurofibromatosis 2 (NF2) and the NF2 gene is mapped to CHR 22, the NF2 gene is a candidate for the meningioma gene. To determine whether NF2 and familial meningioma are allelic mutations, we studied a family with multiple meningiomas and ependymomas in two generations using genetic linkage analysis with DNA markers known to flank the NF2 locus. Multipoint linkage analysis resulted in location scores < -2 for a region of 15 cM including the NF2 region. These results support the existence of a familial meningioma locus that is distinct from the NF2 locus. PMID- 8413973 TI - Reduced basal ganglia volume in HIV-1-associated dementia: results from quantitative neuroimaging. AB - Although brain atrophy is a common neuroradiologic and pathologic finding in patients with HIV-1 infection, especially those with HIV-1-associated dementia complex, it is not clear whether specific regions of the brain are differentially responsible for tissue loss. In this study, we measured volumes of basal ganglia structures on MRIs for three groups: HIV-1-infected homosexual men with HIV-1 associated dementia complex (HIV+ demented), HIV-1-infected homosexual men without HIV dementia (HIV+ nondemented), and noninfected homosexual men. All groups were comparable on age and years of education, and the HIV+ groups were comparable on level of immunosuppression. Total brain volume was smaller in the HIV+ nondemented patients in comparison with HIV- control subjects; the HIV+ demented patients demonstrated even smaller brain volumes than the HIV+ nondemented patients. Smaller basal ganglia volumes, after corrections for intracranial volume, distinguished HIV+ demented patients from the other two groups; there were no differences between the HIV+ nondemented and HIV- groups on basal ganglia volumes. This study suggests that HIV infection causes generalized brain atrophy, but that the clinical features of HIV dementia develop with selective basal ganglia atrophy, consistent with the characterization of HIV dementia as subcortical. PMID- 8413974 TI - Effect of brain irradiation on demyelinating lesions. AB - Demyelinating lesions, such as those in multiple sclerosis, may resemble primary or metastatic brain tumors on CT or MRI, and even be mistaken for neoplasms on biopsy. We encountered five such patients in whom an incorrect diagnosis of CNS neoplasm was made on the basis of radiologic appearance (five) and biopsy (four). All five received radiation therapy, and three chemotherapy. Review of the pathologic findings indicated that the original lesions were not neoplastic but demyelinative. The four patients who received radiation in full tumoricidal doses had unexpectedly poor clinical outcome, suggesting that radiation is especially injurious to patients with demyelinating disease. PMID- 8413975 TI - Crying seizures. AB - We report seven patients with crying during video-EEG-documented simple or complex partial seizures. During simple partial seizures, crying occurred with or without appropriate affect. Crying occurred postictally in two patients and was associated with persistent spiking in one of them. Six patients had ictal activity in the nondominant hemisphere, maximal in the anteromesial temporal region in five and in the mesial frontal region in another. These cases support theories proposing a lateralization of emotion, with right hemisphere dominance for negative affective states. PMID- 8413976 TI - Diagnosis of medial temporal lobe seizure onset: relative specificity and sensitivity of quantitative MRI. AB - Measurement of hippocampal volume by MRI is a new technique with potential value in the localization of epileptic regions, but whether reduced hippocampal volume predicts the location of electrical seizure onset in mixed patient groups is not known. We examined the sensitivity and specificity of this technique among 56 refractory epileptic patients for the diagnosis of medial temporal lobe epilepsy as judged by intracranial EEG recording of spontaneous seizure onset. Since these patients had intracranial EEG because of inconsistent or insufficient localization by noninvasive electrophysiologic, functional, and structural assessment, this patient population can be considered the most difficult to localize. Hippocampal atrophy by MRI volumetry was 75% sensitive to, and 64% specific for, ipsilateral medial temporal lobe seizure onset in this group. Hippocampal atrophy was significantly correlated with longer duration of epilepsy. MRI compared favorably with all other noninvasive means of localization, which had 41 to 73% sensitivity and 45 to 65% specificity to medial temporal ictal onset; of these, none had as acceptable a combination of both adequate sensitivity and specificity. We conclude that MRI volumetry of the hippocampus is a valuable, noninvasive localization method in chronic epilepsy, with a yield and accuracy surpassing all other noninvasive studies used to predict the presence of medial temporal seizure onset. PMID- 8413977 TI - High-dose zidovudine induction in HTLV-I-associated myelopathy: safety and possible efficacy. AB - Ten HTLV-I-associated myelopathy (HAM) patients (four men and six women aged 38 to 58 years) with Expanded Disability Status Scale (EDSS) scores ranging from 4.0 to 8.5 entered an open-label zidovudine study. A high-dosage induction (2 g/d for 4 weeks) was followed by 1 g/d for 20 weeks. Five patients were natives of the Caribbean island Hispaniola, and one each was from Colombia, Cuba, El Salvador, Jamaica, and the United States; all were positive by polymerase chain reaction, and nine had positive Western immunoblots for HTLV-I. Side effects included anxiety, insomnia, gastric upset, anorexia, and loss of taste. Preexisting leg cramps were increased in two and headaches in one. Hemoglobin decreased from a mean of 13.5 to 11.8 g/dl and the hematocrit from 40.7% to 34.9% at 8 weeks, and then stabilized. Neutropenia appeared regularly but did not necessitate drug withdrawal. Mean EDSS scores changed little for the group as a whole, but the seven ambulatory patients improved objectively, with their scores dropping from 5.5 to 4.0 and none worsening. Timed gait improved by at least 50%. Following withdrawal, four of the five who had improved regressed. Zidovudine appears to be safe in subjects with HAM who have no other major health problems and should be investigated further. PMID- 8413978 TI - End-of-dose dystonia in Parkinson's disease. AB - To evaluate the pathogenesis of end-of-dose dystonia in levodopa-treated patients with Parkinson's disease, we discontinued a steady-state optimal-dose levodopa infusion either abruptly or slowly. Although dystonic signs appeared sooner after sudden levodopa termination, in both situations dystonia emerged only when circulating drug levels had fallen to the same concentration and parkinsonian scores had declined by the same amount. Dystonia onset thus appears to reflect the degree, rather than the rate, of reduction in dopaminergic stimulation, and may involve the preferential interaction of dopamine with a receptor subpopulation that does not mediate its antiparkinsonian efficacy. PMID- 8413979 TI - Gangliosides and parkinsonism. AB - The protective effect of GM1 ganglioside against nerve cell death induced by 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was analyzed in monkey mesencephalon. Administration of GM1 before and during MPTP treatment improved motor performances compared with MPTP-treated animals that received saline rather than GM1. Postmortem analysis revealed that GM1 did not protect dopaminergic cell bodies from MPTP intoxication but resulted in an increased immunoreactivity of tyrosine hydroxylase in the perikarya and processes of the surviving neurons. These data suggest that GM1 may be potentially used as a palliative rather than a curative therapy in Parkinson's disease. PMID- 8413980 TI - Effects of bromocriptine on periodic limb movements in human narcolepsy. AB - We studied the effects of bromocriptine on periodic limb movements in sleep (PLMS) of narcoleptic patients. Bromocriptine significantly reduced the number and index of PLMS without improving the nocturnal sleep disruption in that disorder. These results support the hypothesis that dopaminergic mechanisms are involved in the physiopathology of PLMS, and suggest that PLMS do not play a major role in the nocturnal sleep disruption of narcoleptic patients. PMID- 8413981 TI - Interictal heart rate patterns in partial seizure disorders. AB - Epileptogenic mesial temporal damage may alter interictal autonomic patterning. Analysis of heart rate variability in 19 patients with complex partial seizures revealed cases of persistent, high-amplitude, 4 to 9 per minute fluctuations in heart rate during alert waking. This pattern was most pronounced in poor candidates for anterior temporal lobe resection (2/19). The 4 to 9 per minute heart-rate variability pattern may emerge following diffuse, extratemporal, or bilateral mesial temporal damage, which interferes with descending forebrain influences on cardiovascular regulation. PMID- 8413982 TI - Expectancy and response strategy to sensory stimuli. AB - We investigated the relationship between sensory discrimination and the selection of appropriate responses in subjects performing two different reaction-time tasks, in which three auditory stimuli were presented in random order and with a different likelihood of occurrence. Subjects anticipated the need to make different responses based on the likelihood that a particular stimulus would occur on a particular trial. This was determined by the occurrence and distribution of premovement potentials prior to the stimulus, which were consistent with preparation to respond to the most frequently occurring stimulus. These anticipatory cerebral events, however, could be altered after recognition that this frequent stimulus had not occurred. Thus, after the occurrence of a stimulus other than the anticipated frequent tone, the scalp distribution of cerebral potentials changed in a manner suggesting that the next most frequently occurring stimulus was anticipated. Nonetheless, subjects were able to respond to the least probable stimulus both accurately and rapidly despite a failure to anticipate it correctly, as judged by the cerebral "lateralized readiness potential." These results indicate that stimulus evaluation and response selection are integrated and dynamic cerebral processes, and raise doubt about the functional significance of the so-called premovement readiness potential. PMID- 8413983 TI - Guillain-Barre syndrome as the presenting manifestation of hepatitis C infection. PMID- 8413984 TI - Cytomegalovirus-associated transverse myelitis. PMID- 8413985 TI - 'Entomopia': a remarkable case of cerebral polyopia. PMID- 8413986 TI - Bilateral anterior lingual hypogeusia hypesthesia. PMID- 8413987 TI - Adenocarcinoma of the prostate presenting as a third nerve palsy. PMID- 8413988 TI - Ophthalmoparesis due to Burkitt's lymphoma following cardiac transplantation. PMID- 8413989 TI - Leg pain associated with subgluteal lipoma. PMID- 8413990 TI - Mycoplasma pneumoniae infection may cause striatal lesions leading to acute neurologic dysfunction. PMID- 8413991 TI - Disconjugate ocular bobbing. PMID- 8413992 TI - Synovial cyst of the upper cervical spine: MRI with gadolinium. PMID- 8413993 TI - Surgery for PD. PMID- 8413994 TI - Aspartame and seizures. PMID- 8413995 TI - Amantadine withdrawal and neuroleptic malignant syndrome. PMID- 8413996 TI - Lymphocytes in Creutzfeldt-Jakob disease. PMID- 8413997 TI - Giant axonal neuropathy. PMID- 8413998 TI - Hypokalemic periodic paralysis. PMID- 8413999 TI - Permanent global amnesia. PMID- 8414000 TI - Hypertension and seizures. PMID- 8414001 TI - Mononeuritis and cryoglobulins. PMID- 8414002 TI - Vascular dementia. PMID- 8414003 TI - Vascular dementia. PMID- 8414004 TI - Where will future neurologists and neuroscientists come from? PMID- 8414005 TI - Risk factors in young-onset Parkinson's disease. PMID- 8414006 TI - Tension-type and chronic daily headache. PMID- 8414007 TI - Expanding the international classification of seizures to provide localization information. PMID- 8414008 TI - Brain injury by global ischemia and reperfusion: a theoretical perspective on membrane damage and repair. PMID- 8414009 TI - Neurologists--1991 to 1992. Human Resources in Neurology Subcommittee of the American Academy of Neurology. AB - The American Academy of Neurology (AAN) membership survey was developed to define and monitor changing trends in the demography and practice profile of US neurologists. All 11,300 AAN members received a Demographic Information Form (DIF) and 2,600 members also received a Practice Profile Form (PPF) to gather this information. Response rates were 75% from the DIF group and 70% from those who received both. For responding members, age (median, 43 years), sex (83% male), race/ethnic origin (85% white), and medical schools (22% international medical graduates) are similar to those for other physicians in the United States. The ratio of neurologists to population ranges from a low of 1.3 per 100,000 in Wyoming to a high of 11.0 per 100,000 in the District of Columbia. One fourth of neurologists complete at least one residency in addition to neurology and 39% complete a fellowship. Although nearly two-thirds (63%) of neurologists have a full-time or clinical academic appointment, 70% indicate patient care as their primary medical activity. The number of working hours and professional activities of neurologists vary with practice type. Neurologists perform numerous neurodiagnostic tests for reimbursement, particularly neurophysiologic studies and lumbar puncture. One-third of office-based neurologists have an ownership interest in an imaging facility. The most common payment sources for professional fees are commercial insurers (32%) and Medicare (29%), the latter reflecting the large proportion of disabled and elderly treated by neurologists. Office-based neurologists provide 5% charity care and write off an additional 13% of unpaid charges. The practice of neurology is constantly changing.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414010 TI - Nonoperative management of cubital tunnel syndrome: an 8-year prospective study. AB - This study reports the results of a prospective trial of nonoperative management of the cubital tunnel syndrome in 128 patients. Forty-three of the patients had bilateral ulnar nerve compression. At study's end, information was available on 94% of patients and 164 managed extremities, at a mean of 58.6 months of follow up. We scored the degree of severity of cubital tunnel syndrome numerically, based on concepts of the pathophysiology of chronic nerve compression. For statistical purposes, a successful outcome of the nonoperative regimen was not having an operation. Life-table analysis demonstrated that 89% of patients with symptoms only, 67% of patients with abnormal sensorimotor thresholds, and 38% of patients with abnormal sensorimotor innervated density did not have surgery. These differences were significant (p < 0.001) by both parametric and nonparametric analysis. A history of elbow injury significantly worsened outcome (p < 0.02), but the results of the pretreatment electrodiagnosis did not. We conclude that a strict nonoperative regimen be supervised in the initial management of the cubital tunnel syndrome. PMID- 8414011 TI - Headaches in patients with brain tumors: a study of 111 patients. AB - We examined 111 consecutive patients with primary or metastatic brain tumors identified by CT or MRI to characterize brain tumor headache. The median age was 44 years; 34% had primary and 66% metastatic tumors. Headaches were present in 48%, equally for primary and metastatic brain tumors. Headaches were similar to tension-type in 77%, migraine-type in 9%, and other types in 14%. The typical headache was bifrontal but worse ipsilaterally, and was the worst symptom in only 45% of patients. Unlike true tension-type headaches, brain tumor headaches were worse with bending over in 32%, and nausea or vomiting was present in 40% of patients. The "classic" early morning brain tumor headache is uncommon. Nausea, vomiting, an abnormal neurologic examination, or a significant, change in prior headache pattern suggest that the headache may be caused by a tumor. PMID- 8414012 TI - Pathologic correlates of incidental MRI white matter signal hyperintensities. AB - We related the histopathologic changes associated with incidental white matter signal hyperintensities on MRIs from 11 elderly patients (age range, 52 to 82 years) to a descriptive classification for such abnormalities. Punctate, early confluent, and confluent white matter hyperintensities corresponded to increasing severity of ischemic tissue damage, ranging from mild perivascular alterations to large areas with variable loss of fibers, multiple small cavitations, and marked arteriolosclerosis. Microcystic infarcts and patchy rarefaction of myelin were also characteristic for irregular periventricular high signal intensity. Hyperintense periventricular caps and a smooth halo, however, were of nonischemic origin and constituted areas of demyelination associated with subependymal gliosis and discontinuity of the ependymal lining. Based on these findings, our classification appears to reflect both the different etiologies and severities of incidental MRI signal abnormalities, if it is modified to treat irregular periventricular and confluent deep white matter hyperintensities together. PMID- 8414013 TI - Antecedent clinical features associated with dementia in Parkinson's disease. AB - In a prospective cohort study, we examined clinical features in 250 nondemented patients with idiopathic Parkinson's disease (PD) and then evaluated their association with incident dementia during 5 years of follow-up. Seventy-four of the patients became demented. Odds ratios for incident dementia with PD were increased for the following: being older than 70 years of age (2.7; 1.4 to 5.5), having a PD rating scale score greater than 25 (3.0; 1.5 to 6.2), being depressed (2.7; 1.5 to 6.6), being confused or psychotic on levodopa (3.3; 1.3 to 8.7), or having facial masking as a presenting sign (6.1; 1.4 to 26.9). PMID- 8414014 TI - Risk factors for Parkinson's disease. AB - Parkinson's disease (PD) has been associated with rural living, well-water consumption, and pesticide exposure; however, the individual risk contribution of these variables has not been established. We examined social and medical histories of predominantly rural populations to determine relative risk factors for PD. Patients and controls were surveyed regarding residency, occupation, medical history, and social and dietary habits. An initial multiple logistic regression model was confounded by excessive variable colinearity. Principal factor analysis yielded three factors: rural living (including years of rural residency and ground-water use), pesticide use, and male lifestyle (male gender, head trauma, male-dominated occupations). Other variables did not load in factor analysis and were entered separately, with the three factor scores, in a second multiple logistic regression model. Significant predictors of PD emerged (in order of strength): pesticide use, family history of neurologic disease, and history of depression. The predicted probability of PD was 92.3% (odds ratio = 12.0) with all three predictors positive. Pesticide use (distinguishable from rural living) can be considered a risk factor for the development of PD, with family history of neurologic disease and history of depression serving as weaker predictors of PD. PMID- 8414015 TI - Double-blind, placebo-controlled, crossover study of duodenal infusion of levodopa/carbidopa in Parkinson's disease patients with 'on-off' fluctuations. AB - Ten patients with Parkinson's disease suffering severe motor fluctuations completed a double-blind, placebo-controlled, crossover trial of duodenal infusion of levodopa/carbidopa to determine if this technique improved the duration of functional time by reducing plasma levodopa level variability. With infusion, seven patients experienced increased functional "on" hours and decreased number of "off" episodes; however, two patients were slightly worse and one patient experienced no benefit. All 10 patients had significantly decreased variability in levodopa levels permitting better titration of levodopa dosage to individual requirements. Five patients continued to use infusion 12 to 20 months after completion of this study. Selected patients with Parkinson's disease who experience severe motor fluctuations may benefit from duodenal infusion with an improved and prolonged response to medication. PMID- 8414016 TI - The effect of L-dopa infusions with and without phenylalanine challenges in parkinsonian patients: plasma and ventricular CSF L-dopa levels and clinical responses. AB - We monitored the motor response and plasma and ventricular CSF (CSFv) concentrations of L-dopa during IV infusions of L-dopa in two patients with advanced Parkinson's disease. Concentrations of L-dopa in CSFv mirrored, but lagged behind, those in plasma. In the fasting state, the duration, but not the magnitude, of the motor response was greater with increasing plasma and CSFv levels of L-dopa. During IV infusions of L-dopa following oral administration of phenylalanine, a large neutral amino acid that shares a transport system into the brain with L-dopa, the duration of the motor response was markedly attenuated despite undiminished CSFv levels of L-dopa. These observations suggest that either L-dopa entry into CSFv and the brain are differentially affected by phenylalanine or that phenylalanine affects other steps in the motor response. These observations demonstrate that, except in the fasting state, L-dopa in CSFv is not a reliable predictor of motor response. PMID- 8414017 TI - Patterns of neuropsychological performance in Alzheimer's disease patients with and without extrapyramidal signs. AB - We investigated the relationship between extrapyramidal signs (EPS) and the cognitive function in 90 patients meeting NINCDS-ADRDA criteria for probable Alzheimer's disease (AD) by dividing them into those showing no evidence of EPS and those showing at least one EPS on the Unified Parkinson's Disease Rating Scale. Cognitive function was assessed by neuropsychological tests of verbal memory, orientation, abstract reasoning, language, and construction. Patients with EPS were significantly more impaired than patients without EPS on tests of short-term learning and memory, orientation, naming, verbal fluency, and construction, but not on tests of long-term memory, abstract reasoning, or verbal comprehension. These results could not be explained by any differences in age, education, or disease duration between the groups. Since this pattern of neuropsychological impairment resembles that of patients with Parkinson's disease and other EPS syndromes, we hypothesize that EPS are associated with neuropathologic and neuropsychological changes that are superimposed over the classic features of AD. PMID- 8414018 TI - The early course of the Tourette's syndrome clinical spectrum. AB - We retrospectively studied 101 children with Tourette's syndrome to characterize the early course of illness and associated behavioral disturbances of attention deficit hyperactivity disorder (ADHD), obsessive-compulsive disorder (OCD), disruptive behavior (DB), and school problems (SP). For patients without ADHD (45%), OCD (50%), DB (67%), or SP (52%) at the time of initial evaluation, 13% developed ADHD, 8% OCD, 28% DB, and 25% SP during the observation period of 1.6 +/- 1.3 years (range, 0.5 to 7 years). For patients with behavioral disturbances initially, the problems were controlled or resolved for many over time and with therapy: ADHD, 46%; OCD, 47%; DB, 50%; and SP, 67%. Medication changes, assessed after a drug adjustment period between the initial and first follow-up visits (6 +/- 6 months), showed that drug dosages remained largely unchanged and few patients required the addition of new drugs: tic suppressants, 10%; anti obsessional agents, 5%; and stimulants, 12%. Tic suppressants were withdrawn from 12%. PMID- 8414019 TI - Botulinum antibodies in dystonic patients treated with type A botulinum toxin: frequency and significance. AB - We measured serum antibodies to botulinum toxin (ABT) in 96 patients with focal dystonia who had been treated with type A botulinum toxin. The frequency of detectable ABT was 3% (three patients). Patients with ABT had received more than 50 ng of botulinum toxin, and the shortest time between two injections was significantly less than in patients without ABT. The clinical evolution of the three patients was heterogeneous: one had decreased effectiveness with repeated injections, another had persistent improvement, and the third never responded to toxin injections. PMID- 8414020 TI - Prognosis of photoparoxysmal response in nonepileptic patients. AB - Photoparoxysmal response (PPR) is sometimes incidentally encountered in EEGs performed for evaluation of nonepileptic symptoms. We conducted the first long term study of a cohort of nonepileptic patients to determine their risk of having seizures subsequent to incidental recording of PPR. After 6 to 12 years (mean, 9 years), none of the 33 patients had had epileptic seizures. To identify prognostic factors associated with PPR, we performed a case-control study comparing the cohort with 33 age-matched patients who had had epileptic seizures prior to PPR recording. There was no statistically significant difference between the two groups with respect to the clinical or EEG data. Morphology of the PPR discharge was not different between the two groups. Contrary to what is widely believed, persistence of PPR discharges beyond stimulation was not associated with a high risk of developing seizures. Prognosis of PPR is age-dependent and seems favorable in adults without previous epileptic seizures. PMID- 8414021 TI - Mollaret's meningitis associated with herpes simplex type 2 infection. AB - We describe three patients with benign recurrent aseptic meningitis (Mollaret's meningitis). For one of these cases, the episodes of meningitis were associated with herpetic outbreaks. Mollaret cells, which are a hallmark of Mollaret's meningitis, were present in the CSF from two of the three patients. In all cases, herpes simplex virus type 2 DNA was present in the CSF during the acute illness as detected by polymerase chain reaction amplification, although viral cultures from CSF were all negative. Herpesviruses, notorious for frequent and sporadic recurrence, are ideal candidates for the cause of Mollaret's meningitis. PMID- 8414022 TI - A familial syndrome with cutaneous malignant melanoma and cerebral astrocytoma. AB - We describe a family in which cutaneous malignant melanoma or cerebral astrocytoma, or both, developed in eight members over three generations. Other malignancies also occurred with a lesser frequency. In two patients with both malignant melanoma and astrocytoma, the brain tumor followed the diagnosis of melanoma by a period of 2 and 10 years and was the primary cause of morbidity and mortality. The findings in this family may represent a newly described genetic disorder. PMID- 8414023 TI - Thalamic infarctions: differential effects on vestibular function in the roll plane (35 patients). AB - We determined the subjective visual vertical (SVV), ocular torsion (OT), skew deviation, and lateral head tilt in 35 patients with acute thalamic infarctions (14 paramedian, 17 posterolateral, and four anterior polar) and in five patients with mesodiencephalic hemorrhages to obtain the tonic effects on vestibular function in the roll plane. Eight of 14 paramedian infarctions had complete ocular tilt reaction (OTR) with contraversive head tilt, skew deviation, OT, and SVV tilt. The OTR was due to ischemia of the rostral midbrain tegmentum, including the interstitial nucleus of Cajal (INC), and not to thalamic ischemia. Thus, the INC (and the rostral interstitial nucleus of the medial longitudinal fascicle) is the most rostral brainstem structure mediating eye-head coordination in roll. Eleven of 17 posterolateral infarctions exhibited moderate SVV tilts that were either ipsiversive or contraversive. In these 11 cases, vestibular thalamic nuclei (nucleus ventro-oralis intermedius, nucleus ventrocaudalis externus, and nucleus dorsocaudalis) were involved; infarctions in the remaining six were more ventromedial. Anterior polar infarctions did not affect vestibular function in roll. PMID- 8414024 TI - Hereditary congenital nystagmus and gaze-holding failure: the role of the neural integrator. AB - Congenital nystagmus (CN) may be due to an instability of the neural integrator responsible for gaze holding. This longitudinal study tests that hypothesis, investigates the saccadic instabilities of relatives, and assesses the effects of afferent stimulation on both the CN and the coexisting gaze-holding failure. We recorded four siblings who had CN and gaze-holding failure while fixating in primary position and lateral gaze. In lateral gaze, the CN waveforms were superimposed on the centripetal drift caused by the gaze-holding failure; the drift time constants ranged from 300 to 1,450 msec. CN waveforms lacked extended foveation periods. Saccadic instabilities were present in the father and two clinically unaffected siblings; the mother's eye movements were normal. We conclude that CN in the subjects of this study, and in others with idiopathic CN, is not due to gaze-holding abnormalities, and we speculate that development of the fixation reflexes that produce CN foveation periods requires some minimal foveation interval during which the target image is in the foveal area with low retinal slip velocity and acceleration. PMID- 8414025 TI - A dominant hereditary ataxia resembling Machado-Joseph disease in Arnhem Land, Australia. AB - A dominant ataxia among four families of the Arnhem Land Aboriginal people of Northern Australia has many clinical features in common with Machado-Joseph disease. Neuropathology on one case showed multiple system involvement, with sparing of the inferior olives and cerebellar cortex, consistent with Machado Joseph disease. Portuguese ancestry is possible, although not proven. PMID- 8414026 TI - Presymptomatic diagnosis for neurofibromatosis 2 with chromosome 22 markers. AB - Neurofibromatosis 2 (NF2) is a dominantly inherited disorder characterized by multiple tumors of the central nervous system, predominantly bilateral vestibular schwannomas. The gene for NF2 is located in the chromosomal region 22q12 between the loci D22S1 and D22S28. We have performed genetic linkage analysis on 13 NF2 families with a total of nine polymorphic DNA markers, including five which we have recently mapped to this region. Two loci, D22S32 and NEFH, are linked to the NF2 locus at 0% recombination (lod scores of 6.03 and 4.28, respectively). By multipoint linkage analysis, we assign the NF2 gene to an interval of 7 cM, between the loci D22S212 and D22S28. We have used this set of nine markers to construct chromosome 22 haplotypes for the 82 at-risk individuals in this pedigree set. It has been possible to determine, with a high degree of certainty, the carrier status of 70 (85%) of these at-risk individuals. Risk prediction was possible in every case where DNA was available from both parents. Fifty-three of the 70 (76%) informative individuals were assigned decreased risks of being carriers. The use of chromosome 22 probes for risk assessment should result in a greatly reduced number of individuals who require periodic screening for NF2. PMID- 8414027 TI - Prevalence, incidence, and characteristics of multiple sclerosis in Westlock County, Alberta, Canada. AB - We report a prevalence study of multiple sclerosis (MS) in the town of Westlock and surrounding county of Westlock, in Alberta, Canada. The prevalence rate for clinically definite MS on January 1, 1991, was 200/100,000. The average annual incidence rates for patients living in the area at onset were 1.91/100,000 for 1950-1959, 2.85/100,000 for 1960-1969, 3.82/100,000 for 1970-1979, and 7.26/100,000 for 1980-1989. Forty-eight percent of the patients were relapsing remitting. Sixty percent were still walking without assistance. The female-to male ratio was 1.4:1. Mean current age was 47, age at onset 30, and duration of illness 18 years. The majority of patients (40%) experienced multiple symptom onset. Forty percent were of single ethnic origin (primarily British); the remainder were predominantly north European combinations. Twenty-four percent of patients reported another MS relative, six first-degree and one second-degree relative. PMID- 8414028 TI - Pallidoluysian atrophy: dystonia and basal ganglia functional anatomy. AB - The clinical correlates of "pure" pallidoluysian atrophy are not well described. A 59-year-old man presented with 20 years of progressive generalized dystonia, dysarthria, gait disorder, supranuclear vertical gaze palsy, and bradykinesia. At autopsy there was severe bilateral atrophy of the external pallidum and subthalamic nucleus with neuronal loss and marked gliosis. This syndrome may epitomize the consequences of "pure" pallidoluysian atrophy. In this case, dystonia appears to occur in the setting of decreased excitation (increased inhibition) of medial pallidal neurons, a pathophysiologic condition common to several hyperkinetic states. PMID- 8414029 TI - Disruption of automatic speech following a right basal ganglia lesion. AB - Following a right basal ganglia lesion, a right-handed man, age 75, was unable to recite familiar verses. Serial automatic speech, singing, recitation of rhymes, and swearing were impaired, and only idioms and social greetings were preserved. Speech no longer contained overused phrases and he could comprehend automatic speech. In contrast, propositional speech was preserved in both French and Hebrew. Right basal ganglia lesions may impair production but not comprehension of automatic speech. PMID- 8414030 TI - Tuberculous meningitis in the southwest United States: a community-based study. AB - This community-based study analyzed 54 patients with definite or probable tuberculous meningitis (TBM) in New Mexico from 1970 through 1990. Patients ranged in age from 4 months to 86 years. The highest age-specific incidence occurred in the elderly, but 22% of patients were less than 10 years old. Native American patients were overrepresented. Patients were as likely to live in small towns as in large urban cities. Symptoms were present for a median of 13 days before admission. The majority of patients had fevers, headache, stiff neck, and mental changes, such as confusion or lethargy. No patient was admitted comatose. Focal neurologic signs were present in 33%. Laboratory testing found hyponatremia in 79%, pulmonary infiltrates on chest x-ray in 40%, ventricular dilatation on CT or MRI in 52%, and tuberculomas in 16%. PPD skin tests were positive in 64%. CSF cultures grew Mycobacterium tuberculosis in 50%, but colony counts were always lower than 10(2)/ml. As a consequence, acid-fast stains of CSF sediment were reported as positive in only 4%. Six patients were not diagnosed during the hospitalization and died of complications. Twenty-three percent of patients who were appropriately treated also died of complications during the initial hospitalization. Tuberculous meningitis continues to be an important disease in small communities, and affects all ages and ethnic and socioeconomic backgrounds. PMID- 8414031 TI - Incidence and prevalence of myasthenia gravis in western Denmark: 1975 to 1989. AB - We studied the epidemiology of myasthenia gravis (MG) in western Denmark from 1975 to 1989, basing case identification on records from all hospitals in the survey area. The population surveyed was 2.80 million in 1985. The average annual incidence rate was 5.0 per million population (women, 5.9; men, 4.2). The point prevalence rate (January 1, 1990) was 78 per million population (women, 102; men, 53). In men, the incidence increased after 40 years. In women, the incidence rates showed a bimodal pattern with a peak of 7.0 in the age group 20 to 29 years and a second peak of 11.7 in the age group 70 to 79 years. The differences in sex and age-specific incidence rates suggest that younger women are more susceptible to MG than younger men. Old men and postmenopausal women had similar rates. When last examined, 21% of the 220 prevalent cases were in remission and 18% were moderately or severely disabled. PMID- 8414032 TI - The Bereitschaftspotential preceding simple foot movement and initiation of gait in Parkinson's disease. AB - We compared the Bereitschaftspotential preceding a simple foot movement while sitting and a stepping movement while standing in a group of normal subjects and seven patients with mild to moderate Parkinson's disease (PD) while off medication. None had major difficulties stepping to initiate gait. Electromyographic signals from tibialis anterior triggered the averaging of electroencephalographic signals from the scalp. Bereitschaftspotential preceding a standing stepping movement were larger than those before a foot movement while sitting in normal subjects, but no difference was observed in patients with PD. The absence of an increase in the Bereitschaftspotential when stepping in PD may reflect an impairment of the preparation and assembly of the complex sequences of movement necessary to initiate walking, even in the early stages of the illness. PMID- 8414033 TI - Wada memory testing and hippocampal volume measurements in the evaluation for temporal lobectomy. AB - We examined the relationship of Wada memory performance and MRI hippocampal volume measurements to laterality of ultimate seizure localization in 20 patients with complex partial seizures who later underwent temporal lobectomy. Discriminant function analysis employing both Wada memory test asymmetries and hippocampal volume asymmetries correctly classified 100% of the patients into left and right temporal lobe groups. Wada memory asymmetries alone correctly classified 90% of the sample (80% of the sample when the discriminant function included all patients except the one being classified), and hippocampal volume asymmetries alone correctly classified 90% of the patients. A significant correlation was present between Wada memory asymmetries and hippocampal volume asymmetries (r = 0.78), indicating that structural evidence of reduced hippocampal volume has a functional correlate reflected by Wada memory performance. These data suggest that the combination of functional and structural measures is of value in the preoperative evaluation for epilepsy surgery. PMID- 8414034 TI - Detection of hippocampal pathology in intractable partial epilepsy: increased sensitivity with quantitative magnetic resonance T2 relaxometry. AB - Abnormal T2-weighted signal intensity in the hippocampus may be difficult to detect visually, and T2 mapping provides an objective means of assessing signal abnormality. We investigated 50 adult outpatients suffering from intractable partial epilepsy with MRI optimized to detect hippocampal and cortical gray matter abnormalities, and with MR T2 relaxation mapping. The range of normal hippocampal T2 relaxation times is small (99 to 106 msec), and the measurements are reproducible between observers. There were abnormal hippocampal T2 relaxation times in the hippocampus ipsilateral to the site of seizure origin in 70% of patients studied, with the more severe abnormality in the ipsilateral hippocampus in all cases. All hippocampal T2 measurements greater than 116 msec were associated with temporal lobe epilepsy and pathologic or MRI evidence of hippocampal sclerosis, or both. Bilateral abnormalities were present in 29% of cases with hippocampal sclerosis. PMID- 8414035 TI - MRI hippocampal volumes and memory function before and after temporal lobectomy. AB - We investigated the relationship between preoperative MRI hippocampal volumes and clinical neuropsychological memory test data obtained before and after temporal lobectomy and amygdalohippocampectomy for intractable epilepsy in 44 left (LTL) and 36 right (RTL) temporal lobectomy patients. In LTL patients, the difference (right minus left hippocampal volume) between hippocampal volumes (DHF) was significantly (p < 0.001) correlated (r = 0.61) with postoperative verbal memory change as measured by a delayed memory percent retention score from the Wechsler Memory Scale-Revised, Logical Memory subtest. DHF was also positively associated with postoperative memory for abstract geometric designs in LTL patients (r = 0.49, p < 0.005). Resection of a relatively nonatrophic left hippocampus was associated with poorer verbal and visual memory outcome. In RTL patients, larger right adjusted (for total intracranial volume) hippocampal volume was associated with decline in visual-spatial learning, but not memory, following surgery. MRI hippocampal volume data appear to provide meaningful information in evaluating the risk for memory impairment following temporal lobectomy. PMID- 8414036 TI - Nerve conduction studies in Charcot-Marie-Tooth polyneuropathy associated with a segmental duplication of chromosome 17. AB - We evaluated motor conduction velocities in a large group of patients and their unaffected kin from five families in which a segmental duplication of chromosome 17p has shown complete linkage to Charcot-Marie-Tooth disease type 1 (CMT1A). Slowing of conduction was completely concordant with the presence of the segmental duplication; two clinically normal patients had slowed conduction. Nonetheless, among the patients with the CMT1A duplication, conduction velocities varied widely, by > 30 m/sec overall, by > 20 m/sec within families, and often by more than 10 m/sec between siblings and between parents and children. One patient was homozygous for the chromosome 17p duplication and had the slowest conduction velocity observed. Conduction slowing was not age-dependent and was present early in childhood. Our findings demonstrate complete penetrance at an early age of the electrophysiologic phenotype associated with the chromosome 17p duplication and confirm the reliability of nerve conduction studies in establishing the affection status in CMT1A. The great variation in conduction velocity among CMT1A patients emphasizes the influence of factors apart from the shared genetic mutation on phenotypic expression. PMID- 8414037 TI - PCR identification of HIV-1 DNA sequences in brain tissue of patients with AIDS encephalopathy. AB - We analyzed brain tissue from 39 patients for the presence of proviral HIV-1 sequences, using the polymerase chain reaction (PCR) for the amplification of segments of the viral LTR and gag genes. A novel primer extension procedure allowed the detection of a single HIV-1 copy in 1 micrograms DNA. We detected proviral HIV-1 DNA in 16 of 25 brain samples from AIDS patients. Semiquantitative evaluation of the amplified DNAs indicated considerable variation in viral load. Highest levels of proviral DNA were present in brain samples from six patients with clinical evidence of HIV-associated cognitive/motor complex and the histopathologic correlate of HIV leukoencephalopathy or HIV encephalitis. An additional 11 brain samples contained smaller amounts of proviral DNA. In these patients, clinical data were inconclusive regarding the diagnosis of HIV-1 encephalopathy and histopathologically there was no evidence of HIV-1-induced tissue lesions. In nine of 25 seropositive patients with AIDS (36%), brain samples scored negative or did not contain an unequivocal signal indicating the presence of proviral DNA. HIV-1 sequences were not detected in any of 14 control brain samples from HIV-1 seronegative patients. Our data indicate that HIV-1 is present in the central nervous system of the majority (two thirds) of AIDS patients and that the highest levels of proviral DNA in brain tissue are associated with HIV encephalopathy. PMID- 8414038 TI - Sympathetic skin response: age effect. AB - We examined the sympathetic skin response in 100 normal subjects aged 20 to 91 years. The response was present in all younger subjects, but, in the elderly, was present in the lower limbs of only 50% and in the upper limbs of 73%. The amplitude of the response showed a marked decline with age in both the upper and lower limbs. PMID- 8414039 TI - Segmental anhidrosis in the spinal dermatomes in Sjogren's syndrome-associated neuropathy. AB - We describe two women with primary Sjogren's syndrome and sensory neuropathy who had anhidrosis segmentally along the dermatomes of the spinal segment, along with sensory loss. Intradermal administration of cholinergic agents elicited no sweat response in the spinal segments with anhidrosis, whereas a normal response was present in the segments with obvious sweating. These features suggest segmental involvement of the postganglionic sympathetic ganglion cells. PMID- 8414040 TI - Autonomic hyperreflexia in pure progressive autonomic failure: a case report. AB - A 60-year-old woman suffered from recurrent episodes of fever, hypertension, facial flushing, vomiting, stridor, slowly progressive symptoms of hypohidrosis, and orthostatic hypotension. The episodes were synchronous with elevated catecholamine concentration in plasma and urine. This is an example of paroxysmal autonomic hyperreflexia in a setting of pure progressive autonomic failure. PMID- 8414041 TI - Anti-neutrophil cytoplasmic antibodies in vasculitis peripheral neuropathy. AB - Anti-neutrophil cytoplasmic antibodies (ANCA) have been reported to be specific serologic markers of systemic necrotizing vasculitis. We looked for ANCA in 166 consecutive patients referred for evaluation of peripheral neuropathy, wondering if ANCA might be helpful in diagnosing vasculitic neuropathy. ANCA were found in four of six patients with vasculitic neuropathy. However, false-positive results limited the diagnostic usefulness of ANCA in peripheral neuropathy. PMID- 8414042 TI - Positron emission tomography and histopathology in Creutzfeldt-Jakob disease. AB - We studied a 62-year-old man with Creutzfeldt-Jakob disease (CJD), using positron emission tomography (PET) and (18F)-2-fluoro-2-deoxy-D-glucose (FDG). Glucose metabolism was heterogeneously decreased throughout the brain. At autopsy, regional distributions of spongiosis, astrogliosis, and neuronal loss correlated with premortem regional metabolic deficits. These results suggest that PET with FDG may provide metabolic regional markers for CJD neuropathology. PMID- 8414043 TI - Myelitis following chickenpox: a case report. AB - An 18-year-old woman presented with myelopathic symptoms and a T-8 sensory deficit during a primary varicella infection. There were significant MRI changes in both the cervical and thoracic regions. Following treatment with steroids and intravenous acyclovir, the patient improved clinically and the abnormalities on MRI improved. PMID- 8414044 TI - Plasma concentrations of inorganic sulfate in Alzheimer's disease. AB - Recent reports have suggested that plasma concentrations of inorganic sulfate may be lower in patients with Alzheimer's disease (AD). We measured sulfate concentrations in 10 patients with AD and found an average concentration of 0.28 mM, which was not significantly different from the mean concentration in age matched controls (0.32 mM) or young healthy controls (0.27 mM). These results indicate that plasma sulfate concentrations are not altered in AD and that previous reports suggesting altered metabolism of sulfur-containing xenobiotics in neurodegenerative diseases should be reevaluated. PMID- 8414045 TI - Geographic distribution of stroke mortality in the United States: 1939-1941 to 1979-1981. AB - I used data from the National Center for Health Statistics to map the geographic distribution of age-adjusted, race- and gender-specific stroke mortality rates in the United States from 1939-1941 to 1979-1981. Over this interval, stroke mortality rates declined dramatically with convergence of age-adjusted, state specific stroke rates both within and between the various race-gender groups. For each race-gender group, high age-adjusted stroke mortality rates were significantly clustered in the southeastern United States, particularly in the South Atlantic census division, with persistent extreme rates in Georgia and the Carolinas. For whites, low-rate states were concentrated in the Mountain census division and along the northern Atlantic coast. The nonrandom distribution of stroke mortality across the United States, the large magnitude of the difference between high- and low-rate areas, the persistence of the pattern over more than four decades, the similarity of the distribution for different race-gender groups, the lack of delimitation by administrative or political boundaries, and results of national cooperative studies completed in the late 1960s and early 1970s together suggest that the pattern of excess stroke mortality is not an artifact of different diagnostic and reporting practices. Some of the observed geographic variation may be due to both the effects of selective migration and variations in the distributions of stroke risk factors. PMID- 8414046 TI - Dihydroergotamine local reactions reduced by normal saline. PMID- 8414047 TI - Chickenpox stroke in an adult. PMID- 8414048 TI - Natural killer cell leukemia presenting with a peripheral neuropathy. PMID- 8414049 TI - Glossopharyngeal neuralgia from a posterior fossa arteriovenous malformation: resolution following embolization. PMID- 8414050 TI - Carbamazepine-induced urinary retention in long-standing diabetes mellitus. PMID- 8414051 TI - Basilar artery embolism after surgery under general anesthesia: a case report. PMID- 8414052 TI - Neurologic manifestations in Wegener's granulomatosis associated with alpha 1 antitrypsin deficiency. PMID- 8414053 TI - Slowly progressive aphasia. PMID- 8414054 TI - Extrapersonal neglect. PMID- 8414055 TI - Ictal bradycardia. PMID- 8414056 TI - Antiethinyl estradiol antibodies. PMID- 8414057 TI - Anticonvulsant withdrawal. PMID- 8414058 TI - Capsular genu syndrome. PMID- 8414059 TI - Brodmann's numbers. PMID- 8414060 TI - Cocaine-induced stroke. PMID- 8414061 TI - Asymptomatic microemboli. PMID- 8414062 TI - Asymptomatic microemboli. PMID- 8414063 TI - Lyme disease. PMID- 8414064 TI - Chronic fatigue syndrome. PMID- 8414065 TI - ATLS and the Gulf War. PMID- 8414066 TI - ATLS and the Gulf War. PMID- 8414067 TI - Debt and Desert Shield/Storm. PMID- 8414068 TI - Father of urology. PMID- 8414069 TI - Injuries and deaths from collecting war souvenirs in Operation Desert Storm. AB - Data were collected on 22 patients who sustained injuries or death from collecting war souvenirs during Operation Desert Storm from December 1990 to May 1991. Nine of 16 patients injured and 3 of 6 patients killed were non-combat arms personnel. Non-combat personnel closely followed combat units, and subsequently were exposed to unexploded ordnance. Combat and non-combat personnel should be warned that innocent-appearing material on or near a battlefield may be an explosive device. They should be instructed concerning the types of ordnance used, as well as the dangers of unexploded ordnance. PMID- 8414070 TI - A descriptive analysis of wounds among U.S. Marines treated at second-echelon facilities in the Kuwaiti theater of operations. AB - Medical data from 120 U.S. Marine Corps trauma admissions to second-echelon facilities during Operation Desert Shield/Storm were examined. Sixty-five percent of the admissions occurred between February 22 and February 28, the time frame corresponding to the ground war and the preliminary mobilization period. Penetrating wounds were the most prevalent types of injury, followed by lacerations, open fractures, and closed fractures. The most frequent anatomical regions sustaining injuries were the leg, head, hand, and arm. Fragments were the causative agent of 63% of the admissions that had this variable recorded, while gunfire was the cause in 20% of the cases. The median injury-to-admission interval increased from 0.67 hours in the non-ground war period to 4.41 during the ground war. PMID- 8414071 TI - Patient care and nursing practice when staff requirements exceed staff availability. AB - This study examined the Workload Management System for Nurses at a tertiary-care Army hospital to determine the incongruence between recommended nursing care hours and actual nursing care hours provided. The purpose of the study was to describe patient care and nursing practice when calculated staff requirements exceed actual staff availabilty. The findings of the study indicated that basic nursing care tasks were accomplished; however, professional development activities were sacrificed. The data reveal that nurses do not have the time to grow professionally through research or education, and they are reduced to assembly-line mentality as they go from task to task without being able to care for a patient as a person. PMID- 8414072 TI - Psychological and war stress symptoms among deployed and non-deployed reservists following the Persian Gulf War. AB - The present study examined the incidence of war-related psychological distress among Persian Gulf War veterans. A total of 591 Army, Navy, and Marine reservists were administered the Mississippi PTSD Scale, the Beck Depression Inventory, and the SCL-90R. Combat-deployed reservists showed significantly higher levels of psychological symptomatology that non-deployed reservists, generally corresponding to levels of stress exposure. No significant effects were found for race or prior combat exposure, but significant differences were found between combat-deployed male and female reservists. Despite the brevity and the outcome of the Persian Gulf War, significant symptomatology exists among this population. PMID- 8414073 TI - Radial keratotomy in the soldier-aviator. AB - Radial keratotomy (RK) is an ophthalmological procedure that alters the shape of the cornea, making it "flatter," causing the desired shift to farsightedness. Complications can be minor and "normal" in the immediate post-operative period, or can include problems that occur in many eyes, that persist but do not decrease best corrected visual acuity, or may include events that potentially or actually threaten vision and may produce blindness. At best, only 50% of patients can expect to have 20/20 uncorrected vision 2-3 years after surgery. Diurnal fluctuations in vision (two-five Snellen lines) can persist years after RK. Perhaps 1% of patients may have a two-three Snellen line loss of best corrected vision or ghost images that interfere with clear vision. Disabling glare can disrupt daily activities. Eyes that have undergone RK are at increased risk of corneal rupture after blunt eye trauma. The visual demands of the active duty military, and more dramatically the military aviator, are incompatible with RK. Therefore, RK should not be performed on active duty soldiers, nor should enlistees be accepted if they have undergone the procedure. PMID- 8414074 TI - Positive and negative symptoms of schizophrenia as predictors of length of military service: a retrospective study. AB - Forty-three chronic schizophrenic veterans had negative symptom, positive symptom, and smooth pursuit eye movement evaluations to determine if the severity of impairment in these dimensions could retrospectively predict their length of military service. For the entire patient group, there was a strong trend for severity of negative symptoms to predict the length of military service (p = 0.055). However, different associations for these variables emerged when the patients were divided into groups with more or less than 2 years of service. The length of military service might serve as a type of "filter" that meaningfully separates out different subtypes of schizophrenic veterans. PMID- 8414075 TI - Revitalizing division mental health in Garrison: a post-Desert Storm perspective. AB - The Division Mental Health Section (DMHS) has long been an essential component of the combat mental health mission. Relatively little attention has focused on its important functions in garrison. The authors, both veterans of Operation Desert Storm, review the DMHS mission in garrison and highlight forces currently jeopardizing the success of that mission. They describe Persian Gulf lessons with important implications for garrison mental health care and then discuss possible solutions to the varied problems facing DMHSs. They conclude, most importantly, that an effective DMHS must identify with the division, sharpen soldier and officer skills, respect organizational boundaries, and continuously prepare for combat. PMID- 8414076 TI - Utilization of portable ventilators in combat zone hospitals. PMID- 8414077 TI - Debriefing: a therapeutic intervention. AB - An injured soldier may not have the benefit of the mental health interventions available to other combatants. A team of psychiatric liaison nurses report on the therapeutic effects of debriefing in the treatment of the traumatically injured soldier. Debriefing not only affords an opportunity for the patient to share thoughts and feelings about the trauma, but provides additional data regarding effective coping mechanisms. This information also assists the health care team to identify individuals at risk for developing post-traumatic stress disorder. Patient teaching can then be directed toward helping each individual recognize the signs and symptoms of stress unique to him/her and mobilize a plan for obtaining assistance. Debriefing also assists the treatment team in identifying a soldier's specific information deficits regarding the system, i.e., medical boards and convalescent leaves. PMID- 8414078 TI - The constipated serviceman: prevalence among deployed U.S. troops. AB - The prevalence of constipation in deployed servicemen was determined in a sample of military personnel aboard the USS Iwo Jima LPH 2 during Operation Desert Shield. Results were obtained from a bowel function questionnaire issued to 500 deployed marines and sailors. When constipation is defined as no bowel movement for greater than 3 days, 3.9% of the Marine/sailor personnel are constipated when in their home environment as compared to 6.0% when they are aboard ship and 30.2% while in the field. Alternatively, when constipation is defined as the presence of certain anorectal complaints (hard stools, straining, painful defecation, and bleeding with defecation), the incidence is 7.2% when at home as compared to 10.4% aboard ship and 34.1% in the field. These results confirm that whether constipation is defined as infrequent bowel movements or presence of symptoms of constipation, significantly more servicemen will be constipated when in the field as compared to their home environment. Since approximately one-third of Navy/Marine Corps personnel deployed in a field environment will be constipated, preventive measures ought to be evaluated. PMID- 8414079 TI - Drafting a parenteral nutrition order form utilizing a physician assessment process. AB - The utilization of a physician critique process in the design of a parenteral nutrition order form, encouraging a standard order option, is assessed. In the critique process, physicians from surgery, medicine, and medical intensive care teams were tasked to use the draft order form and guideline sheet to prescribe a parenteral nutrition order for a hypothetical patient. Prescriber recommendations and problems with the nutrition order form and guideline sheet directed editorial improvements. The physician assessment process is an effective means to measure utility of a draft parenteral order form and prescribing guideline sheet. By utilizing physician expertise, deficiencies inherent in such an order form may be corrected prior to implementation. PMID- 8414080 TI - Refractive error distribution and incidence among U.S. Army aviators. AB - Spectacle incompatibility has been a major problem in the fielding of advanced Army avionic and electro-optical systems. As a result, routine contact lens wear may be instituted as an option to spectacles. Refractive error data were extracted from the Aviation Epidemiological Data Registry, a computer-accessible repository of flight physical medical information on the entire Army aviation population. Refractive error distribution patterns in the class 1 and class 2 flight physical populations were analyzed in order to provide a contact lens supply reference database, estimate the annual incidence of refractive error development, and estimate costs of possible spectacle-wearer flight-duty deselection. Contact lens wear appears to be a more cost-effective alternative than deselection. PMID- 8414081 TI - Common chronic disease in Vietnam-era veterans and nonveterans. AB - Little research has been conducted investigating the general experience of military service during Vietnam and its long-term relation to common nonpsychiatric problems. The relationship between Vietnam-era veteran status and common chronic disease was examined using data from the 1987 National Health Interview Survey. Compared to a cohort of nonveterans, Vietnam-era veterans were not more likely to have common chronic disease. Moreover, veterans were not significantly more likely than nonveterans to have limitations in activity. The findings indicate that military service during the Vietnam conflict does not place veterans at greater risk for common chronic disease. PMID- 8414082 TI - Protective ability of the standard U.S. Military Personal Armor System, Ground Troops (PASGT) fragmentation vest against common small arms projectiles. AB - This study analyzes the ballistic protection provided by the standard U.S. Military Personal Armor System, Ground Troops fragmentation vest against threats from handgun and shotgun projectiles. Damage assessment was conducted using 10% ordnance gelatin, a tissue simulant which has been proven to have a close correlation with living tissue. The standard fragmentation vest was shown to provide protection from commonly encountered handgun bullets and shotgun pellets, but not from shotgun slugs or center-fire rifle bullets. PMID- 8414083 TI - Primary closure of gunshot wounds caused by high-velocity rifles. AB - Sixty gunshot injuries in 48 patients were treated in Mobile Surgical Hospital (30-bed capacity) of the Mountain and Commando Brigade in Hakkari, Turkiye, between May 1990 and May 1991. The David Sisk method was used for the classification of the injuries. Eleven type 1 injuries were treated conservatively, 39 type 2 and 10 type 3 injuries were treated by extensive debridement with suction drainage and primary closure. No infection was seen in type 1 injuries. The infection rate was 2.56% in type 2 and 20% in type 3 injuries. The following results were obtained: type 1 injuries must be treated by conservative measures; type 2 injuries must be treated by primary closure following extensive debridement and suction drainage; type 3 injuries can be treated by extensive debridement with suction drainage and primary closure. PMID- 8414084 TI - The effects of litter carrying on rifle shooting. AB - This study investigated whether the use of a shoulder harness would affect shooting accuracy after patient litter carrying. Two- and four-person teams, 12 male and 9 female soldiers, fired at targets before and after (1) a 15-minute bout of rapid, short litter carries and lifts, and (2) a moderate speed 30-minute litter carry with and without a harness for both types of carries. Shooting accuracy was 10% poorer (p < 0.05) after the 15-minute bout (mean +/- SD = 8.9 +/ 1.9 mm) than after the 30-minute carry (8.1 +/- 1.7 mm). Four-person teams using litter-carriage harnesses had 17% tighter shot groups (45.5 +/- 30.4 mm2) (p < 0.05) than four-person teams that did not use harnesses (54.5 +/- 26.1 mm2) and two-person teams with (56.3 +/- 29.1 mm2) or without harnesses (54.9 +/- 30.7 mm2). The harness can potentially improve shooting accuracy after litter carrying. PMID- 8414085 TI - The maturing of MEDRETEs. AB - The Medical Readiness Training Exercise (MEDRETE) is generally regarded as an effective tool for enhancement of U.S. foreign policy in developing nations. However, if MEDRETEs are evaluated with parameters other than emotion, there is found to be little durable effect and virtually no improvement in the health status of the host nation. The classic MEDRETE is ineffective, costly, and provides only an evanescent benefit to the served population. It should be replaced by PMRETEs (Preventive Medicine Readiness Training Exercises) targeting preventive medicine concerns which would provide low-cost, durable contributions to the health status of growing nations. PMID- 8414086 TI - Orthopedic manifestations of hyperlipoproteinemia: an unusual case of knee pain. PMID- 8414087 TI - Diagnosis of superior sulcus tumors: a further use of the thoracoscope. AB - Most irradiation therapists require a tissue diagnosis of a superior sulcus tumor prior to initiating preoperative irradiation. In the rare case, obtaining a tissue diagnosis via the normally used procedures proves challenging. A technique for obtaining a tissue diagnosis using a thoracoscope in this circumstance is presented. PMID- 8414088 TI - [The thermodilution determination of right ventricular volumes and ejection fraction in the critical patient. Volumetric vs pressure measurement assessment]. AB - The widespread use of hemodynamic investigation techniques, allowed a better understanding of the right ventricle (RV) pathophysiology and led to progressive reevaluation of its role. A modification of the classical Swan-Ganz catheter, made possible the measurement of the ejection fraction (EF) and of the end systolic and end-diastolic volumes of RV just by the simple application of the thermodilution technique. In this paper, we first refreshed the basic theoretical principles of the technique and then presented our preliminary results of one year experience in ICU. A non-homogeneous group of 36 critically ill patients (septic shock 17, COPD 13 and ARDS 6) was studied. Specifically we found that neither the data of central venous pressure nor those of RV end diastolic pressure, were able to estimate the real preload, i.e. RV end diastolic volume (r = 0.01 and r = 0.03 respectively with "p" not significant). We compared the data of RV EF with that of the end systolic pressure/volume (P/V) ratio in a group of patients before and during the administration of dobutamine to evaluate their sensitivity to identify changes of contractility. The results obtained support the superiority of RV end systolic P/V ratio over RV EF to detect variations of contractile status. We conclude that the use of the thermodilution technique to measure the volumes of RV, allows a real evaluation of the preload. At the same time it avoids all the problems associated with the measurement of transmural pressure and with the changes produced by shifting of intrapleural pressure. Finally the end systolic volume may be combined with the pressure data to estimate the contractile status and, in our experience, this parameter has proved more sensitive than EF in order to detect changes of contractility of the right ventricle. PMID- 8414089 TI - [End-tidal CO2 as a predictive index of regional perfusion and its relation to aortic flow. A clinical study during peripheral vascular surgery]. AB - Using a new haemodynamic monitoring system, we prospectively measured the end tidal carbon dioxide (ETCO2) and the aortic blood flow (ABF) in 7 patients undergoing major vascular surgery to evaluate the usefulness of ETCO2 as a predictive indicator of regional blood flow. Previous studies demonstrated a high correlation between ETCO2 and Cardiac Output during CPR (r = 0.79), this allows us to conclude that ETCO2, under conditions of constant ventilation, reflects the circulatory status. We investigated the relationship between ETCO2 and ABF, and our observations confirmed that the two parameters undergo highly significant variations (p < 0.001), but low correlation was found (r = 0.15), so the increase in ETCO2 after aortic declamping depended on re-perfusion of ischaemic regions. The ETCO2 concentration increased immediately in 6 patients after declamping (p < 0.05). In 1 patient, the increase wasn't significant, but he underwent a new operation because of malfunction of the prosthesis. Our findings suggest that ETCO2 monitoring may provide clinically useful information about regional perfusion that can be used to guide therapy. PMID- 8414090 TI - [Plasmapheresis and high-dose immunoglobulins in the treatment of acute demyelinating polyneuropathy]. AB - The authors report two clinical cases of Guillain-Barre syndrome, treated with contemporaneous use of plasmapheresis and high dose gammaglobulin. An early diagnosis and immediate application of such a therapeutic association, are of paramount importance in order to obtain remission of the neurologic symptomatology within a short time. They report the therapies carried out to avoid complications connected with this syndrome as well as cause which led to quit steroid use in these patients. PMID- 8414091 TI - [Combined anesthesia (epidural-general) in thoracic surgery: the cardiocirculatory response to induction. Propofol vs midazolam plus alfentanil]. AB - Thoracic epidural block may unpredictably blunt the physiological sympathetic response to anaesthetic agents and profound hypotension could result at induction time. The authors evaluate two different induction regimens in 16 patients scheduled for major thoracic surgery in order to assess which of them would produce less hemodynamic changes. After the institution of high thoracic epidural block (6 ml lidocaine-CO2 2% + epinephrine 1/200,000 level: th.2-3), patients were randomly allocated in two induction groups: P and M. Group P received 2 mg/kg of propofol over 30 seconds; group M received midazolam 0.2 mg/kg plus alfentanil 30 mcg/kg. In Both groups induction was followed by a significant drop in blood pressure (-18% in group M and -37% i group P. In P group the fall in blood pressure is more pronounced than M group (p < 0.05). Noteworthy in both groups cardiac frequency do not increase and in M group significantly lowers. In the authors view the midazolam plus alfentanil induction technique compare well with propofol alone and would recommend its use in poor risk patients. PMID- 8414092 TI - [The treatment of postoperative shivering with nefopam hydrochloride]. AB - After having considered the physiopathological mechanism of shivering, the authors have estimated in a randomised study the effects of the resolution of shivering by nefopam hydrochloride. This drug a dose of 0.20 mg/kg resolved the symptoms in 100% of cases and the placebo hasn't resolved a case so that case it also obtained the same effects. The authors have proved the efficacy of the nefopam hydrochloride in the control of postoperative shivering in the recommended dose. PMID- 8414093 TI - [Intraoperative awareness: a true or false problem?]. AB - The authors base their remark on a few cases of awareness which have been reported in their hospital and on the history a patient who experienced intraoperative awareness during cesarean section. Attention is focused on the ethical, social and medico-legal aspects of this phenomenon. Awareness may severely affect the patient's psychic equilibrium with the consequent onset of severe neuroses leading to the possibility of legal action and claims for compensation. Clinical experience has shown that the use of halogenate anesthetics, even in low concentrations, may help to avoid this phenomenon. Correct relations between the anesthetist and patient before and after surgery may also, even in the event of awareness, reduce the incidence and severity of consequent psychological sequelae. PMID- 8414094 TI - [The organizational problems of the hospitals of the Nord Italia Transplant Program (NITp) engaged in the activities of organ retrieval for transplantation. The Collaborative Group of Resuscitation Anesthetists]. AB - If the quality of results of organ transplantation in NITp is highly satisfactory, the same cannot be said for the number of transplants performed, which only cover 20-25% of the requirement. To understand the causes of organ shortage, a study group of Anaesthesiologists and Transplant Coordinators from the North Italy Transplant Program (NITp) investigated, through a questionnaire addressed to the Heads of 103 Intensive Care Units (ICU) in 92 Hospitals in the NITp area, some of the organizational problems linked to donor identification and treatment and to organ retrieval. The questionnaire took into consideration the number of possible donors identified in 1990, those retrieved and the causes for non retrieving organs and examined a number of variables linked both to retrieval and to ICUs and Hospitals organization. The results show that potential donors were 461: 143 (31%) were used, 138 (30%) were lost due to family opposition to organ donation, 192 (20%) for clinical reasons and 88 (19%) for organizational reasons. The latter figure represents 5% of non retrieval in the most active ICUs and increases to 50% in the Hospitals that had procured no donors in 1990. The main obstacles for ICUs to procure organ donors are: convey the Medical-Legal Committee, carry out of complicated administrative procedures, availability of round the clock specialized equipment for neurological assessment (especially for ICU outside Neurosurgical and Neurological departments), inadequate number of medical and nursing staff, most of all at night, and finally the difficulty in the management of non-traumatic cerebrovascular patients. PMID- 8414095 TI - [The use of subarachnoid bupivacaine analgesia in malignant hyperthermia susceptible subjects. A clinical case report]. AB - The authors report a case of malignant hyperthermia in susceptible woman undergoing safenectomy. Spinal anesthesia with bupivacaine was performed after dantrolene prophylaxis per os. No complications occurred during and after surgery. This case confirms the safety of local anesthetics and it suggests that anaesthesia should not be refused to caffeine halothane contracture test positive patients. PMID- 8414096 TI - [Vascular surgery and vascular surgeons: yesterday and today]. PMID- 8414097 TI - [Benign tumors of the liver]. AB - Diagnostic and therapeutic findings of benign hepatic tumors are analysed. In particularly the authors describe the more frequent tumors such as angioma, adenoma and FNH, but also give a guidelines on how to approach less frequent lesions. PMID- 8414098 TI - [Acute cholecystitis in videolaparoscopy]. AB - The authors describe their experience in 15 cases of acute cholecystitis treated with laparoscopic cholecystectomy in emergency. Only in one patient, affected with an abscess of the upper right abdominal space, a laparotomy was performed. The results were excellent. The 14 patients treated with laparoscopic cholecystectomy were quickly discharged from the hospital (on an average 4 days after the surgical treatment) and the convalescence period was very short (range 10-15 days). In the patient treated with laparotomic cholecystectomy the postoperatory period was regular. PMID- 8414099 TI - [Post-traumatic hernia of the diaphragm]. AB - The paper reports the series of post-traumatic hernias of the diaphragm observed during the period between 1954 and June 1992. This group of 77 patients (59 males and 18 females) were aged between 6 and 75 years old. Both acute and chronic hernias were reported; in 60 cases the diaphragmatic lesion was on the left, in 9 cases on the right, and there were 8 cases of laceration to both hemidiaphragms. The most frequently herniated organ was the stomach, followed by the transverse colon and spleen. With regard to the access route used, the authors draw attention to the fact that during the first twenty years of the series thoracotomies were prevalently used, whereas during the last decade laparotomy has been used in cases of early post-traumatic hernia, thoracotomy in cases of chronic post-traumatic hernia or if there is suspected damage to organs and intrathoracic structures. PMID- 8414100 TI - [Cardiorespiratory changes induced in man by the acute occlusion of the main bronchus]. AB - Following the exclusion of the ventilation of a lung, perfusion is arrested as a consequence of the Von Euler and Liljestrand vasoconstrictive reflex. However it is as yet unknown how long this phenomenon takes to manifest itself in human beings. In order to verify whether hypoxic pulmonary vasoconstriction follows immediately on the arrest of ventilation or whether there is one some delay, in 21 candidates for pneumonectomy a bronchial block was provoked during fibrobronchoscopy and the hemogasanalytic modifications induced by the acute occlusion of the main bronchus have been studied. The analysis of the resulting data allows for the hypothesis that the exclusion of lung ventilation for 14 minutes is not sufficient to cause the complete manifestation of the hypoxic vasoconstrictive reflex. PMID- 8414101 TI - [Prognostic significance of cell-mediated immune response in patients with colorectal carcinoma. Preliminary results]. AB - Pre-operative skin test response has been evaluated in 50 cases of colorectal carcinoma. Cellular defense's depression was correlated with tumoral staging (p = < 0.001). In B2 and C groups, a higher incidence of metastatic and local recurrences was registered in subjects with low response. These last are suitable for a randomized trial including immunotherapy. PMID- 8414102 TI - [Infarction of the sigmoid. Clinical considerations]. AB - The authors present a case of sigma infarction. They examine the etiopathogenesis, underline the clinical and instrumental signs and discurs the vascular anatomy and diagnostic difficulties. Operation is considered to be necessary to reduce the severe of prognosis. PMID- 8414103 TI - [Progressive pneumoperitoneum in the surgical treatment of conspicuous ventral hernias. Importance of the procedure to restore a good respiratory function]. AB - A recent clinical report regarding a patient with a voluminous laparocele treated using progressive pneumoperitoneum justifies this short paper on the use of this method as a preparatory act to reparative surgery. The paper emphasises the undoubted advantages deriving from the use of this technique, especially in the case of those subjects with a particularly poor respiratory function secondary to hernia. PMID- 8414104 TI - [Spigelian hernia. Clinical and anatomo-surgical considerations]. AB - The authors describe a case of Spigelian hernia observed and treated. This hernia is uncommon, although the true incidence is probably greater than the small number of patients reported in literature. The signs and symptoms of the hernia are not always characteristic and then a correct diagnosis can be difficult. Sometime instrumental examination is essential for diagnosis, when clinical examination is not clear. Surgical repair as mandatory for a correct reconstruction of the abdominal wall and a prevention of recurrences. PMID- 8414105 TI - [Pneumatosis cystoides intestinalis. Etiopathogenic and clinical considerations regarding a case]. AB - Pneumatosis cystoides intestinalis is a rare condition characterized by gas filled cysts in the submucosa and/or subserosa of the intestine. The authors present a case of Pneumatosis cystoides intestinalis occurring in a patient with pyloric stenosis by duodenal ulcer and report a wide review of the literature. They discuss the major areas of study in its etiology, histopathology, diagnosis and treatment. PMID- 8414106 TI - [Eosinophilic granuloma of the maxillary bone. Review of the literature and report of a case of mandibular location in an HIV-positive patient]. AB - Following a short review of the literature, the authors report a case of an eosinophilic granuloma in bone in an immunodepressed patient (manifest AIDS) suffering from chronic hepatitis B and C. The possible link between the viral infection and immunodepression and the pathology in question in then discussed. PMID- 8414107 TI - [Angiomatoid malignant fibrous histiocytoma. Clinico-pathologic and immunohistochemical study of a case]. AB - The results of a clinico-pathologic and immunohistochemical study of an angiomatoid malignant fibrous histiocytoma are reported. This lesion is an uncommon tumor of the superficial soft tissue, of low-grade malignancy, typical of adolescence and early adult life. The patient, a 10-year-old female, presented with a mass of the left popliteal fossa, treated with surgical excision of the tumor and the surrounding cutaneous and subcutaneous tissue. The tumor was a well circumscribed, firm nodule measuring 2.5 x 1.0 cm. Histologically, it showed aggregates of spindled and rounded cells often lining cystic cavities filled with blood. The immunohistochemical analysis revealed a cytoplasmatic immunoreactivity for KP1 (CD68), which was taken as indicating that the tumoral mesenchymal cells had acquired phagocytic capacities. The patient is well without signs of local recurrence or metastatic disease 4 years after the surgical treatment. The case reported confirms that appropriate local surgery is the elective therapy for this type of soft tissue tumor. PMID- 8414108 TI - [Ethical problems in the treatment of breast carcinoma]. PMID- 8414109 TI - Endoscopic treatment of malignant dysphagia. AB - Palliative endoscopic treatment of dysphagia in patients with inoperable oesophageal cancer includes: dilation, Nd:YAG laser photocoagulation and intubation, used alone or in combination. Such procedures are usually performed on an outpatient basis and are associated with a low rate of morbidity and mortality. From 1978 to 1988, 476 patients (401 males, 75 females) were treated at the Endoscopy division of the National Cancer Institute of Milan for inoperable primary or recurrent malignancies of the oesophagus or cardia or for extra-oesophageal neoplasms causing dysphagia. Dilation was used in 172 cases, Nd:YAG laser photocoagulation in 90, prosthesis insertion in 72, dilation and laser in 97, and prosthesis and laser in 45. Functional improvement was reported in 75% of patients after dilation, in 89% after laser treatment, in 80% after intubation, in 80% after dilation and photocoagulation, and in 89% after laser and intubation. The median duration of dysphagia-free interval was 4 weeks in dilated patients, 6-8 weeks in photocoagulated patients and 20 weeks intubate patients. Overall median survival was 6.2 months. The complication rate was: 1.4% in dilation treatment, 1.4% in laser photocoagulation, and 8.8% in prosthesis intubation. Mortality related to endoscopic treatment was 2.1% (10/476 patients). Relief of dysphagia is one of the most important goals of palliative treatment in patients with inoperable oesophageal neoplasms. Moreover, endoscopic palliation improves the quality of life in the patients, with a low complication rate. PMID- 8414110 TI - [The operative stages for the reconstruction of perineal stoma continence. Our technique]. AB - The authors outline the stages of abdominoperineal surgery due to rectal cancer to achieve the reconstruction of perineal-stomic continence. The operation is simple to perform, without particular vascular risks and sepsis; it does not necessitate temporary abdominal colostomy and, lastly, does not oblige postoperative stomic functional rehabilitation. PMID- 8414111 TI - [Surgical alternatives in the treatment of perforated duodenal ulcer]. AB - The therapy of perforated gastroduodenal ulcer is still discussed, especially today that good drugs and new surgical devices are available for the treatment of this disease. The authors try to establish a valid protocol for the diagnosis and the therapy of patients affected with this disease. PMID- 8414112 TI - [Current surgical approach in the treatment of pancreatic pseudocysts]. AB - INTRODUCTION: The paper evaluates the role of surgery in the treatment of pancreatic pseudocysts in the light of current knowledge. The authors underline the important part played by ultrasonography and CT in monitoring the evolution of pseudocysts: ETG on account of its high levels of sensitivity and specificity and CT due to its diagnostic accuracy which is equivalent to almost 100%; these methods allow the surgeon to decide if and when to operate. MATERIALS AND METHODS: A series of operations were carried out by the authors' division in 15 patients suffering from pancreatic pseudocysts. Emphasis is placed on the high percentage of emergency operations, 40%, due to complicated pseudocysts. External drainage was always used in emergency surgery, whereas in elective surgery it was preferred to use cystojejunostomy with a Roux-en-Y loop, followed by cystogastrostomy; cystoduodenostomy was only chosen in one case. The operative mortality rate was 20%. DISCUSSION: In elective surgery the operation which currently gives the best results is cystojejunostomy with a Roux-en-Y loop, but cystogastrostomy should not be overlooked due to the ease with which it is performed. A recent acquisition is the use of needle aspiration and CT- or ETG guided percutaneous drainage in the treatment of post-necrotic pseudocysts, therefore after 4-8 weeks' maturity, both in a rapid-growth from and in the event of suppuration. Pancreatic resection is indicated for multiple atypical pseudocysts caused by retention, and this also serves to treat the underlying pancreopathy. Duodenocephalopancreatectomy is now considered to be excessive by the majority of authors. In emergency surgery, drainage of the pseudocysts is the method of choice using either a surgical or percutaneous method. PMID- 8414113 TI - [Surgical treatment of non-toxic multinodular goitre. Retrospective evaluation of a personal case series]. AB - The authors base their study on a retrospective evaluation of a personal series of patients and on a review of the literature on this topic. They focus their attention on surgical problems relating to the type of treatment of non-toxic multinodular goitre. On the basis of the data obtained, they underline that in this pathology total thyroidectomy is the most appropriate form of surgery and does not entail a higher morbidity in relation to more conservative resection. PMID- 8414114 TI - [Cardiac echinococcosis. Case report and review of the literature]. AB - Cardiac echinococcosis is a rare disease. We report the case of a patient affected by cardiac echinococcosis who underwent surgical treatment successfully. A forty year old woman was hospitalized referring palpitation and dyspnea. The patient had undergone surgical pericystectomy of the right lobe of the liver for echinococcosis 6 month before. Chest X-ray film showed a round opacity well delineated on the left side of the heart, Ghedini reaction was negative, ECG was normal. After a review of literature we analyze anatomo-clinic and therapeutic aspects of the disease. PMID- 8414115 TI - [Two cases of chronic constipation in the adult: Hirschsprung disease and idiopathic megacolon]. AB - Two clinical cases of megacolon with long-standing constipation in the adult are presented: the former associated with Hirschprung's disease and the latter idiopathic. Diagnostic procedures and surgical treatment are described. The different criteria of diagnosis and therapy are discussed after a literature review. PMID- 8414116 TI - [Primary non-Hodgkin's lymphoma of the breast. Clinico-pathologic study of 2 cases]. AB - The authors describe two cases of malignant non-Hodgkin's lymphoma in a mammary site observed at IRCCS Policlinico San Matteo (Pavia). Histologic, immunohistochemical and clinical features are illustrated. Correct diagnosis is essential so that appropriate multidisciplinary treatment may be applied. PMID- 8414117 TI - [Foreign body decubitus: unusual cause of acute appendicitis]. AB - The authors describe the origin and the physiopathological events in acute appendicitis. They show an uncommon case of acute appendicitis. The aetiology was an opaque radiation foreign body causing a subacute abdominal disease. At operation the authors discovered the above body free in the abdominal cavity; the appendix was not in ischemic phase but completely cut off bycecal plant. They suppose aetiology to be of a decubitus origin. PMID- 8414118 TI - [Pneumoperitoneum without perforation]. AB - Usually, pneumoperitoneum is due to perforation of a hollow abdominal organ. In the absence of peritoneal symptoms, pneumoperitoneum may be due to other causes. Two patients with pneumoperitoneum without perforation were treated in our department. A 41-year-old female patient was admitted to our hospital with abdominal pain for 12 hours, without vomiting. On abdominal examination fullness of the liver was absent, the abdomen was not distended and there were no signs of peritoneal irritation. The roentgenogram showed free air under the diaphragm. Laparotomy was performed because of the abdominal complaints. No perforation could be found. A 70-year-old female patient was admitted to the hospital because of mild abdominal pain. For a week she had a severe attack of coughing due to bronchitis. Big capacity of air had been blown in the stomach without abnormalities in the roentgenologic examination. The treatment was conservative. Both patients left the hospital in good health. More knowledge of the less frequent causes of pneumoperitoneun without perforation can possibly contribute to to refraining from exploratory laparotomy in these cases. PMID- 8414119 TI - [Starch granulomatous peritonitis. Description of a clinical case]. AB - The starch peritonitis is a rare disease which is present in the patients recently submitted to a laparotomy. The ethology is an allergic reaction to the surgical glove starch. The symptoms are a continuous temperature, a leucocytosis and a eosinophilia. Macroscopically there is a micronodular dissemination of peritoneum with adhesion and a limited ascitis. Microscopically, there are gigantocellular granulomas with birifrangent inclusions to polarizing light. A diagnosis is difficult before surgery. The laparoscopy and the microscopic examination of ascitis can give a piece of information. The therapy is medical. PMID- 8414120 TI - [Large retroperitoneal hemangiopericytoma. Presentation of a clinical case]. AB - Hemangiopericytoma is a rare and solitary solid tumor originating from pericytes, which are contractile pericapillary cells, first identified and described by Zimmermann in 1923. This neoplasm occurs most commonly in the lower extremities and retroperitoneum. The Authors describe the case of a patient affected with a giant retroperitoneal hemangiopericytoma. Echographic and computed tomographic scans have been fundamental in defining location and size of the neoplasm as well as its relations to the surrounding structures. However a definitive diagnosis was possible only by histologic examination, supported by the proper immunohistochemical stains. The Authors emphasize the difficulty to reliably predict the biological behaviour of these tumors and therefore the necessity of a follow-up of the patients for at least five years, even in the case of a histologically "benign" hemangiopericytoma. PMID- 8414121 TI - Popularity, friendship, and emotional adjustment during early adolescence. PMID- 8414122 TI - Adolescents and their friends. PMID- 8414123 TI - Conflict management among close peers. PMID- 8414124 TI - Close friendships in early and middle adolescence: typology and friendship reasoning. PMID- 8414125 TI - Close friendship and imaginary companions in adolescence. PMID- 8414126 TI - Theory is not a four-letter word: needed directions in the study of adolescent friendships. PMID- 8414127 TI - [Didanosine in the treatment of HIV infection]. PMID- 8414128 TI - [International collaboration among firms for the development of drugs for AIDS. An agreement among 15 pharmaceutical firms]. PMID- 8414129 TI - [Neuroimmunomodulation. The bi- and unilateral correlations between the nervous system and the immune system]. AB - The nervous system-immune system relationships, namely the modulatory effects of the former upon the latter and vice versa, have been considered both in physiological and pathological conditions. The nervous system communicates with the immune system in two biochemical ways, by means of neurohormones and neurotransmitters, using anatomical and functional pathways related to the pituitary system (the manifold endocrine axes between hypophysis and peripheral endocrine glands) and autonomic system (especially the orthosympathetic one, and to much lesser extent the parasympathetic system). The immune system is able to modulate some nervous functions via cytokines, peptidic molecules produced by immunocytes, that can also be secreted in certain conditions by neural cells, such as astrocytes, microgliocytes and even neurons. The knowledge of physiology of the neuroimmunomodulation and related pathological changes may be very useful for understanding the pathogenesis and the pathophysiology of neuroimmunological diseases, and for their diagnosis and treatment. PMID- 8414130 TI - [The incidence, etiology and clinical significance of visceral mycoses in patients with AIDS]. AB - The incidence, aetiology and clinical significance of visceral mycoses in HIV infected subjects were evaluated by a retrospective survey of the clinical and microbiological records of 237 consecutive AIDS patients followed-up since 1984. Seventy-four patients out of 237 (31.2%) (56 males, 18 females; 55 IV drug abusers, 7 heterosexuals, 6 homobisexuals, 3 blood recipients and 3 children with congenitally-acquired HIV infection) presented 77 different episodes of visceral fungal infection as a whole, represented by candidiasis in 56 cases (oesophageal 45, pulmonary 5, sepsis 2, eye involvement 2, endocarditis and invasive oropharyngeal infection in the remaining 2 patients), cryptococcosis in 17 cases (meningoencephalitis in all subjects, with disseminated infection in 11 of them), and aspergillosis in 4 cases (pulmonary 2, cerebral and cranio-facial in the remaining 2 patients). In 57 out of 74 patients (77%), visceral mycoses were diagnostic or concurrent with the diagnosis of AIDS. Fungal diseases, as a whole, showed a significantly higher incidence (p < 0.03) among drug abusers, whereas homobisexual men presented a significantly lower frequency (p < 0.001, chi-square test) than AIDS patients with other risk factors for HIV infection. The onset of cryptococcosis was significantly associated with the male sex (p < 0.005, Fisher exact test). All subjects suffering from a visceral mycosis were severely immunosuppressed, with a higher rate of neutropenia in patients developing Candida and Aspergillus spp. infection (23 out of 56 patients with visceral candidiasis and 3 out of 4 cases of aspergillosis had an absolute neutrophil count lower than 1500 cells/mm3), while a severe reduction in CD4+ lymphocyte count was more evident among patients with cryptococcosis (13 out of 17 patients had a CD4+ cell count lower than 50/mm3). After remission of the primary episode of fungal infection (obtained in 80.5% of cases), the incidence of relapse observed in a long follow-up period (mean time 57.6 +/- 39.2 weeks) was elevated both for patients with cryptococcosis (7 cases out of 17) and subjects with candidiasis (19 cases out of 53), with no significant difference among patients receiving a secondary prophylaxis or not (22 relapses observed in 53 patients treated with maintenance antifungals versus 4 episodes in 8 patients followed for a comparable mean time with no antimycotic treatment). Fifty-two out of 74 patients (70.3%) have died up to now; in 21 of them death was due to or associated with the visceral mycosis (cryptococcosis in 11 cases, candidiasis in 8, aspergillosis in 2).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8414131 TI - [Invasive thermometry and prognostic thermometry in radiological hyperthermia]. AB - The term radiological hyperthermia is used to describe the exposure of biological tissues to temperatures just above physiological ones, namely between 42 and 45 degrees C. A machine for hyperthermia (SAPIC SV03 produced by Aeritalia, Caselle, Turin) is been experimentally used at the authors' institute. The present study aims to analyse the technical and qualitative aspects of temperature control using invasive and previsional thermometry. The term invasive thermometry is used to describe the insertion of small teflon catheters into the biological body through which thermometric probes are introduced to control deep temperature. Previsional thermometry is the phrase used to describe the computer simulation of hyperthermic treatment, starting from knowledge of the tissue to be heated and the type of applicator used. The simulated treatment and the real temperature measured are then compared in order to optimize the treatment used. PMID- 8414132 TI - Platelet aggregation in presence of anticoagulants dependent pseudothrombocytopenia. AB - We report a case of pseudothrombocytopenia anticoagulants dependence in a healthy woman. Platelet count was performed on the automated impedance haematological analyzer utilizing peripheral blood samples anticoagulated with ethylene diamine tetra-acetate, heparin and sodium citrate. We pointed out that the severe thrombocytopenia was principally time and ethylene diamine tetra-acetate dependent. As regard both the temperature (37 degrees C) and the other anticoagulants (heparin, sodium citrate), the phenomenon was variable. The phenomenon "in vitro" was confirmed by the normal aggregation, moreover we showed that the platelets of a normal subject aggregated with plasma and ethylene diamine tetra-acetate of pseudothrombocytopenic subject. We report this case because often, in a healthy subject, it is possible to make a mistake in diagnosis and to employ more sophisticated and expansive investigations. Moreover it is very important to point out the pseudothrombocytopenia in case of heparinic therapy because it is possible to have a dilated thrombocytopenia. PMID- 8414133 TI - [Congestive gastropathy in liver cirrhosis]. AB - Congestive gastropathy has emerged as a new nosological entity that can be included among the complications of advanced liver cirrhosis. It has been defined as the macroscopic changes of gastric mucosa occurring in portal hypertension that are associated with vascular mucosal and submucosal dilatation and ectasia without significant inflammatory changes. The pathogenesis of congestive gastropathy has not been completely cleared up. Many epidemiological and clinical studies and some tests on animals lead most Authors to think that the cause of this disease is a chronic increase of pressure in the portal vascular system. However the involvement of humoral factors cannot be excluded as, for example, the presence of high plasma levels of gastrin and histamine or a decrease of E2 prostaglandin in the gastric mucosa. The macroscopic lesions typical of congestive gastropathy can be seen through endoscopy. Up to now mosaic-like pattern, red points, cherry-red and black-brown spots and erosions have been observed. These changes are prominent in the area near the gastric body and cardias, but can be present in all parts of the stomach. The frequently reported spontaneous bleeding corresponding to cherry-red spots make the presence of these lesions to be considered a sign of severe congestive gastropathy. The prevalence of congestive gastropathy in cirrhotic patients is between 30% and 70%. This condition is more frequent in patients with large esophageal varices and severe liver disease and in patients submitted to endoscopic variceal sclerotherapy. Congestive gastropathy is a frequent cause of acute and chronic bleeding: 10-20% of gastrointestinal bleeding episodes occurring in cirrhotic patients are caused by this condition and about 30% of cirrhotics with portal hypertension will have one or more acute bleeding in a four year follow-up. The percentage of subjects with chronic hemorrhage in the same period can reach 90%. At the moment is not possible to suggest a therapy able to prevent or cure the acute or chronic bleeding associated with congestive gastropathy. beta-blockers seem to be a promising treatment. However, further and larger clinical trials are necessary to settle definitively their efficacy. PMID- 8414134 TI - [The importance of the follow-up in patients operated on for breast cancer. A retrospective analysis of 2482 cases]. AB - INTRODUCTION: The aim of the follow-up in breast cancer patients is the early detection of recurrences and the diagnosis of second tumours to improve the survival rate and/or the life quality of patients. MATERIAL AND METHODS: In this study we analyze retrospectively 2482 cases, treated in the period 1964-1988; all patients underwent surgery: mastectomy in 1979 cases and conservative surgery in 503 cases. The follow-up was performed by clinical and radiological (chest X-ray, bone scintigraphic scanning, echotomography of the liver and mammography) examinations every 3-4 months in the first and second year, every 6 months for 5 years and then every year. The duration of the observation time ranges from 42 to 330 months (median 172). RESULTS: Relapses were detected in 928 cases (37.4%): 289 loco-regional recurrences (11.6%) and 639 distant metastases (25.8%). The 62.1% and the 92.5% of the failures appeared after 3 years and 10 years follow-up respectively. Local recurrences were observed in 14.1% of all relapses after mastectomy and in 9.3% after conservative surgery. 36.3% and 88.1% of the local failures appeared after 2 years and 5 years follow-up respectively. Regional lymph nodal recurrence occurred in 14.2% of all relapses: 1.6% in the axilla, 9.8% in the supraclavicular region, 2.8% in the mediastinal nodes. Distant metastases were observed in 68.9% of all relapses: 31.3% in the bone, 17.6% in the lung and 8.1% in the liver. An increase of the serum markers (CEA, TPA, CA 15 3) appeared 2-10 months before other clinical or radiological signs of disease; CA 15-3 showed the best predictive positive value. Metachronous tumours in the contralateral breast were detected in 103 cases (4.1%), all staged as T1-T2. The modalities of diagnosis of the relapses were studied in 350 patients. In these cases 76% of recurrences were detected by history (62%) and physical examination (14%), 10% by chest X-ray, 12% by bone scan and 2% by liver echotomography. 36% of relapsed patients were asymptomatic and the mean anticipation of diagnosis was 4 months. An increase of survival was observed only in patients with loco regional recurrences. CONCLUSIONS: The results obtained in our study and the literature data confirm the importance of long-term follow-up in the breast cancer patients. Clinical examination and mammography represent the most important modalities of diagnosis for the local and distant relapses and the second neoplasms in the contralateral breast. PMID- 8414135 TI - [The diagnosis of non-Hodgkin's lymphoma in a patient with Marfan's syndrome]. AB - The Marfan syndrome is classified as a heritable disorder of connective tissue whose pathogenetic mechanism has not yet been defined. An inborn error of protein metabolism, particularly in collagen has long been hypothesized, but conclusive evidence on the matter is not available. It is usually diagnosed in young patients and can be associated with other disease. We report a case of an unusual association with a malignant non-Hodgkin lymphoma in a 55-year-old patient, never described before. PMID- 8414136 TI - [Tertiary hyperparathyroidism during chronic kidney failure under dialysis treatment. Apropos a clinical case]. AB - The authors describes a case of tertiary hyperparathyroidism (HPTH) in a uremic patient on intermittent dialysis treatment: the term refers to an adenoma with ensuing uncontrolled parathormone (PTH) secretion rate arising on the ground of hyperstimulated hypertrophied parathyroid glands. The syndrome was heralded clinically by bone pain, psychiatric disorder and biochemically by increased levels of calcium and alkaline phosphatase (AP), while parathormone (PTH), did not change from basal very high levels as commonly found in uraemic patients. As hypercalcemia in the hemodialyzed is an infrequent finding the only alternative explanation could have been hypercalcemic secondary HPTH related to hyperplastic autonomous parathyroids. For no clinical and laboratory findings as well as US findings and double scintigraphy (99mTc and 201mTl) may suggest differential diagnosis the patient underwent total parathyroidectomy which actually revealed an adenoma of the left superior parathyroid gland. Bone pain and psychiatric disturbances disappeared and now get well on chronic dialysis treatment and 1.25 OH Vit D3 supplement. PMID- 8414137 TI - [Gianni Bonadonna receives the Bristol Myers Squibb award for cancer research]. PMID- 8414138 TI - [Invasive procedure in obstetrics: a journey in the field of fetal medicine. Four videocasettes proposed by the Italian Association for the Study of Malformations and by "Recordati" (new)]. PMID- 8414139 TI - [Serum beta hCG levels and transvaginal echography in the early phases of pregnancy]. AB - One hundred thirty two patients at an early gestational age were monitored every other day to establish beta hCG levels at which the gestational sac, the yolk sac and fetal heart motion can be sonographically detected. These structures were observed at significantly lower beta hCG levels by means of the vaginal probe in comparison with the abdominal one. In normal pregnancy a gestational sac of 1-3 mm was detected at a mean hCG level of 1150 UI/l (range 800-1500); the yolk sac was detected at a mean hCG level of 6000 UI/l (range 4500-7500); fetal hart motion was visible at a mean hCG level of 10,425 UI/l (range 8650-12,200). The yolk sac and fetal heart motion were always detected when the gestational sac was greater than 11 and 17 mm respectively. A practical method to evaluate early first trimester pregnancy is suggested. PMID- 8414140 TI - [Transvaginal sonography as a screening method for the identification of patients at risk of postmenopausal endometrial pathology]. AB - Starting from the anatomopathological assumption that endometrial thickness in postmenopausal women never exceeds 3 mm, and in view of the reliability of measurements made using an echographic probe, the authors evaluated the value of transvaginosonography (TVS) as a mass screening method for postmenopausal endometrial pathologies. A group of 74 patients were examined who were recruited from those attending the out-patient menopause clinic. All subjects conformed to the following admission criteria: amenorrhea for the past two years; absence of oestroprogestin therapy for at least six months; absence of vaginal blood loss. A 5 MHz probe was used to measure maximum endometrial thickness on the longitudinal plane; values were divided by two if the surfaces were adjacent. Patients were monitored according to the following protocol: endometrial thickness under 1 mm control every 12 months; thickness between 1-3 mm--control every 3 months; thickness equal to or over 4 mm--hysteroscopy, targeted biopsy, possible scraping and TVS control after 3 months. The group was subdivided as follows: 65 patients (87.8%) were without risk; 3 patients (4%) belonged to the intermediate risk group; 6 patients (8.2%) belonged to the high risk group. Of the latter, 4 revealed an endometrial polyp, one presented uterine polymyomatosis and one a proliferative-type endometrium. The authors' experience is still limited but the absence of false positives encourages them to continue their research using this simple and well tolerated method. It might represent a valid alternative to hysteroscopy as a screening method in the asymptomatic population at risk for endometrial carcinoma. PMID- 8414141 TI - [Use of transvaginal ultrasonography in the early diagnosis of endometrial neoplasia]. AB - The paper examined the efficacy of transvaginal sonography in measuring the endometrium in menopausal women. The study included 50 women, both asymptomatic and with metrorrhagia, all of whom had been in menopause for at least 1 year and were under 75 years old. In symptomatic patients sonographic findings were compared with histological tests. Histological correlation was only performed in asymptomatic patients with endometrial findings of over 8 mm or in those undergoing surgery for other pathologies. Sonography revealed a good level of correlation with histological findings. Endometrial lines measuring less than 8 mm had no pathological significance. On the contrary, in those cases over 8 mm it was important to evaluate the overall morphological significance of sonographic findings and for this purpose histological diagnosis was necessary. Ten hyperplastic forms and 2 endometrial adenocarcinoma were diagnosed in this group. In terms of the sonographic image the hyperplastic forms appeared as a thick, highly reflective and well defined endometrial layer; on the contrary, carcinoma appeared as an irregular, highly reflective area with the loss of the hypoechogenic peripheral halo. The authors conclude that this easily performed and minimally invasive method is of value as as screening method for endometrial carcinoma and should be recommended for women receiving replacement therapy. PMID- 8414142 TI - [Asymptomatic peritoneal endometriosis following cesarean section]. AB - Endometriotic peritoneal implants of small dimensions were discovered in five patients undergoing laparoscopic examination for diseases unrelated to endometriosis. All patients were asymptomatic for endometriosis and each had undergone at least one previous cesarean section. Biopsies taken were studied by light microscopy which evidenced variably predominating epithelial or stromal components of lesions Perls' method was used to evidence tissue ferric iron resulting from long-standing micro-haemorrhagic foci. From this study, isolated, asymptomatic peritoneal implants of endometrium are suspected to occur frequently after cesarean section although the progression of this condition to a symptomatic state is considered infrequent and independent of specific histologic aspects of lesions. PMID- 8414143 TI - [Fetal macrosomia. Maternal and fetal implications]. AB - Ninety-six cases of fetal macrosomia have been tested to focus on the factors necessary to select women running the risk of delivering macrosomic fetuses. The results of these studies are the following ones: a) pregnant women with pathology are multiparas; b) they are usually characterized by peculiar weight increase in pregnancy; c) they have gone beyond pregnancy term; d) they have previously delivered macrosomic fetuses. The Authors focus on the necessity to anticipate diagnosis and to plan caesarean section in order to avoid the traumatic lesions this kind of babies are frequently affected by. PMID- 8414144 TI - [Risk factors of endometrial carcinoma]. AB - The authors base their experience on 12 women affected by endometrial carcinoma. Problems have been diagnosed in the post-menopausal and climacterium period, considering the clinical, hormonal and lipidic profile and some tumoral markers. The most risk factors for the endometrial adenocarcinoma have been collected in the adiposity with hypertension and hyperglycemia, above all in nullipara women with late menopause and dyslipidemia. PMID- 8414145 TI - [Primary dysmenorrhea treated with sublingual piroxicam]. AB - Open non comparative study for the evaluation of the efficacy and tolerability of the new piroxicam formulation, the Fast Dissolving Dosage Form tablets for sublingual administration in the treatment of primary dysmenorrhea. The 54 patients enrolled in the study have been treated with piroxicam sublingual tablets: 40 mg single daily dose for the first two days and 20 mg for the following three days for a total of 5 days of therapy to be repeated for two consecutive menstrual cycles. The drug efficacy was evaluated on the basis of variation of intensity in pain and associated symptoms, as cephalgia, nausea, vomiting, etc. The ability to perform normal daily activity has also been evaluated. The intensity of spontaneous pain in the first and second cycles showed a statistically significant improvement (p = 0.0001) only 15 minutes after drug administration. This improvement increased in the first and in the following days. The relief from pain was stronger in the second cycle, in fact none of the patients had to assume a further analgesic drug, as happened during the first cycle of therapy. As regards the symptoms associated to pain, as cephalgia and low back pain, they significatively decreased starting from the first cycle of treatment. The improvement became more marked during the second cycle of therapy. Local and systemic tolerability was good. Only 5 patients experienced systemic side effects in the first cycle, and 12 in the second cycle. Three patients experienced local side effects in the first cycle, and 1 in the second cycle. These effects were well tolerated and did not cause the treatment discontinuation. Moreover, most of the side effects occurred, as nausea and diarrhoea, are symptoms commonly associated to dysmenorrhea. In conclusion, piroxicam fast dissolving dosage form for sublingual administration, in the treatment of primary dysmenorrhea, showed its analgesic efficacy 15 minutes after the drug administration. It has also a good local and systemic tolerability. PMID- 8414146 TI - [Fluconazole in the treatment of vaginitis caused by Candida albicans]. AB - The authors verified the local activity and tolerance of the antimycotic drug Fluconazole in 125 cases of women suffering from clinical diagnosed vulvovaginitis caused by Candida albicans. Results were positive with regard to the efficacy and safety of the treatment. PMID- 8414147 TI - [Use of seat belts during pregnancy]. AB - After a review of the world literature on the advantages and compulsiveness of seat belts, the authors recommend pregnant women to use them. Moreover, they invite gynaecologists and midwives to favour their use during psycho-prophylaxis courses. PMID- 8414148 TI - [Prevalence of endemic goiter in Sicilian pregnant women in the Alte Madonie region]. AB - The "Alte Madonie" is a hill-mountain community in Sicily with a population of approximately 30,000 inhabitants. It includes the towns of Alimena, Bompietro, Blufi, Castellana Sicula, Fasano, Gangi, Geraci, Petralia Sottana, Soprana and Polizzi generosa. This is an area where goitre is endemic. Between January 1990 and September 1991 out of a population of 400 pregnant women examined at various stages of pregnancy, goitre was found in 1 out of 7.8 women (51 cases). 74.5% (38 cases) of those with goitre had undergone endocrinological tests, in all except two cases, a long time before the start of pregnancy. Three pregnant women (5.8%) were receiving suppressive therapy with L-thyroxine. 58.8% (30 cases) had undergone hormonal controls and laboratory data had revealed cases of hypothyroidism; in the remaining cases hypothyroidism had been excluded on the basis of medical examination. One patient, not included in this series who had received IVG, had been treated a few months before with strumectomy and radio iodio-therapy for non-differentiated carcinoma which had developed on the goitre. In conclusion, these findings underline the high frequency of goitre in pregnant women from the "Alte Madonie", and the need for greater care in the prevention and treatment of this pathology. There is a clear need for integrated health education programmes and early diagnosis to be carried out in schools and work places. The role played by the gynecologist who first sees the pregnant women with goitre is vitally important, especially in the case of those patients which dedicate little time to the health of their own bodies either for cultural reasons or due to the demands of work both inside and outside the family. PMID- 8414149 TI - Molecular genetics of Alport syndrome: the clinical consequences. PMID- 8414150 TI - ACE inhibition: antihypertensive treatment of choice in progressive chronic renal failure? PMID- 8414151 TI - Adhesion molecules: receptors dictating local immune responses. PMID- 8414152 TI - Chronic renal failure in India. AB - In a series of 2028 patients with chronic renal failure, the diseases leading to renal failure, the presence or absence of reversible factors and their nature, and the rate of decline of renal function of the most common conditions have been described and analysed. Seven diseases: chronic interstitial nephritis (27.85%), diabetic nephropathy (26.76%), chronic glomerulonephritis (18.20%), benign nephrosclerosis (10.06%), chronic pyelonephritis (7.29%), focal glomerulosclerosis (3.20%), and autosomal dominant polycystic disease of the kidneys (2.07%), accounted for 95.43% of all the patients. These diseases were studied in greater detail and the results are presented here. It was found that there was a great variation in the rate of decline of renal function in the different groups, with chronic glomerulonephritis and focal glomerular sclerosis progressing most rapidly, diabetic nephropathy slightly slower, and the others at a less alarming pace. However, once serum creatinine had reached 177 mumol/l there was an inexorable decline in renal function and the end stage was reached in almost all patients. PMID- 8414153 TI - Hereditary nephritis (Alport's syndrome)--clinical profile and inheritance in 28 kindreds. AB - Sixty-three patients, (52 males and 11 females) from 28 kindreds of hereditary nephritis (Alport's syndrome) were identified over a 14-year period from 1977 to 1991. Group I included 51 patients with (a) positive family history of haematuria with or without chronic renal failure, (b) characteristic GBM changes on electron microscopy, (c) characteristic ocular signs, and (d) high-frequency sensorineural deafness. Group II included 12 patients with a negative family history. All of them had evidence of renal disease with characteristic ocular signs and deafness and four had characteristic GBM changes on electron-microscopy. The main clinical features were haematuria in 96.8%, deafness in 82.5%, and diminished visual acuity in 66.7% of affected subjects. Hypertension was present in 71.4% patients. Pure tone audiometry revealed high-frequency sensorineural deafness in 96.8%. Ocular examination showed bilateral anterior lenticonus in 37.8%, retinal flecks in 22.2%, cataract in 20%, and keratoconus in 6.7% patients. Proteinuria (> 2.0 g/24 h) was detected in 31.8%. Sixteen (57.1%) of the 28 index patients (all males) were diagnosed for the first time when they presented with end-stage renal disease. Serum creatinine in the overall group ranged from 0.9 to 18.7 mg/dl(7.81 +/- 5.37 mg/dl). Adequate renal tissue was obtained by biopsy in 14 patients. Light-microscopy revealed focal segmental glomerulosclerosis in five, mesangial proliferation in four, chronic interstitial nephritis in three, and mesangiocapillary and crescentic glomerulonephritis in one each. Electron microscopy showed characteristic changes in the GBM in seven specimens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414154 TI - IgG subclasses of antineutrophil cytoplasm autoantibodies (ANCA). AB - Sera that had been positive in routine ELISA for ANCA were studied retrospectively for the IgG subclass distribution of these autoantibodies. An ELISA previously developed for measurement of IgG subclasses of anti-GBM antibodies was modified for this purpose. Of a total of 247 sera, 114 were found to be positive in at least one of the assays for IgG subclasses of anti proteinase 3, 72 of these patients were men and 42 were women, giving a ratio of 1.8. Also 134 sera were positive in at least one of the IgG subclass assays for antimyeloperoxidase (MPO), with a male/female ratio of 0.97. The ANCA seem to consist mainly of IgG1 and IgG4 autoantibodies. Among the anti-MPO group, IgG2 is relatively common and IgG3 is scarce. Contrasting with this, IgG3 is relatively common in the antiproteinase 3 group. In this group high IgG2 titres are rare. Twelve sera were found to be positive for both autoantigens. Clinical data were studied for 44 patients. Prognosis for old patients was found to be poor. Patients with inactive disease were often positive in only one subclass assay, while patients with active disease were positive in two or more subclass assays (P < 0.01). PMID- 8414155 TI - Antigen size influences the type of glomerular pathology in chronic serum sickness. AB - Chronic serum sickness was induced in four groups of Wistar rats by immunization with BSA, cationized BSA (cBSA), human IgG (HuIgG), or human IgM (HuIgM), followed by repeated intraperitoneal (i.p.) injection of the antigen used, to study the effect of the characteristics of an antigen on renal immunopathology. Renal tissue sampled 2, 4, 7, and 9 weeks after the start of the i.p. injections was examined by light-, immunofluorescence-, and electron-microscopy. Proteinuria was measured in urine collected over 24 h. All animals given BSA, cBSA, or HuIgG developed progressive renal disease characterized by initial deposition of antigen and rat Ig in the mesangium of rats given BSA or HuIgG, and minimal amounts in those given cBSA, followed by the appearance in the first instance of subendothelial deposits in the animals receiving BSA or HuIgG, and later subepithelial deposits in those given BSA, cBSA, or HuIgG. The appearance of immunoglobulin deposits along the glomerular capillary wall was associated with the onset of massive proteinuria reaching average levels of 450 mg/24 h for rats given BSA or cBSA, and 500 mg/24 h for those given HuIgG. Animals injected with HuIgM showed only mesangial deposits of human IgM and rat Ig without the development of proteinuria. Under light-microscopy, rats given BSA, cBSA, or HuIgM showed minimal abnormalities, whereas those receiving HuIgG showed transient but severe influx of granulocytes in glomeruli with the development of diffuse proliferative glomerulonephritis in association with a long-lasting phase characterized by subendothelially localized immune aggregates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414156 TI - Exaggerated natriuresis induced by sodium chloride infusion in essential hypertension is accompanied by an exaggerated urinary 3' 5' guanosine monophosphate excretion. AB - The effects of an intravenous infusion of physiological saline on plasma atrial natriuretic peptide (ANP), guanosine 3' 5' monophosphate (cGMP) concentrations, and on urinary cGMP and sodium excretion were studied in 13 patients with essential hypertension, class I according to WHO criteria, and in 10 healthy subjects. It was found that the groups did not differ as to basal and infusion induced plasma ANP and cGMP and basal urinary cGMP and sodium excretion, but the sodium chloride infusion resulted in a significantly greater urinary cGMP and sodium excretion and creatinine clearance in hypertensive than in control subjects. The results of this study demonstrate that patients with essential hypertension respond to an intravenous sodium chloride load not only with exaggerated natriuresis, but also with augmented urinary cGMP excretion. The latter finding may in part be due to a greater glomerular filtration of cGMP, but increased renal contribution cannot be excluded. Apart from the possible stronger intrarenal effect of ANP on cGMP production in patients with hypertension, independent direct effect of volume expansion on cGMP excretion and modified activity of other cGMP generating systems may all be responsible for the higher urinary cGMP excretion in essential hypertension. PMID- 8414157 TI - Determination of total body water in uraemic patients by bioelectrical impedance. AB - The measurement of total body water by bioelectrical impedance in a group of renal patients was evaluated against the tritium dilution method. The effect of haemodialysis and the presence of peritoneal dialysate on the impedance were also investigated. The correlation between the two methods is r = 0.90 with a residual standard deviation of 3.7. The standard deviation of the differences between the two methods against the means was 3.66 which means that total body water (TBW) estimated by the bioelectrical impedance (BEI) method may be 6.18 l (X +/- 2 SD) above or 8.38 l below the 3H2O method. The BEI method overestimated the actual weight loss after haemodialysis (3.87 +/- 1.7 l versus 2.43 +/- 1.8 l) but underestimated the volume of peritoneal dialysate in situ. The BEI method would not be appropriate for use in assessing total body water and monitoring acute volume changes in patients with renal failure who are on strict fluid restriction. PMID- 8414158 TI - High-dose 15-deoxyspergualin monotherapy surpasses methylprednisolone in its therapeutic effect on advanced lupus nephritis in New Zealand black/white F1 hybrid mice, and low-dose combination may be synergistic. AB - To compare the therapeutic effect of 15-deoxyspergualin with that of methylprednisolone on advanced lupus nephropathy of New Zealand black/white F1 hybrid (B/W) mice, and also to study the possible synergistic effect of both drugs, B/W mice were heminephrectomized at 32 weeks of age, and were divided into six groups. Each group of mice was treated with 50 microliters phosphate-buffered saline (PBS), 3 mg/kg methylprednisolone, 20 mg methylprednisolone, 0.6 mg DSP, 6 mg DSP, or with 3 mg methylprednisolone plus 0.6 mg DSP, s.c., four times per week for 8 weeks. Urine and blood samples (by tail vein venipuncture), as well as renal tissue specimens, were taken at 32 and 40 weeks of age. The degree of proteinuria, and serum anti-DNA activity (by ELISA) were determined. Renal specimens were evaluated with light- and immunofluorescence (C3)-microscopy, the degree of pathological changes being semi-quantitated and expressed as total light-microscopy (LM) and immunofluorescence (IF) scores. The survival rate at 40 weeks of age was significantly elevated in 0.6 mg DSP, 6 mg DSP, and methylprednisolone + DSP groups of mice compared with the control group. The appearance rate of significant post-treatment proteinuria was comparable among all groups. The difference (post-treatment titre--pretreatment titre) of serum anti-DNA activity in the 6 mg DSP and methylprednisolone + DSP groups were significantly less, while that of the 3 mg methylprednisolone group was greater compared with the control level. As for the total LM score, the levels significantly decreased in the 6 mg DSP, methylprednisolone + DSP and 3 mg methylprednisolone groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414159 TI - Long-term follow-up study of 268 diabetic patients undergoing haemodialysis, with special attention to visual acuity and heterogeneity. AB - We studied the long-term outcome of 268 patients suffering from diabetic end stage renal disease (DM-ESRD) treated with long-term haemodialysis between 1978 and 1991, with special emphasis on visual acuity as well as the heterogeneity of DM-ESRD. The 50% patient survival on haemodialysis was 60 months. Visual disturbances were found in 73.1% (392/536) of eyes at the start of haemodialysis. Chronological assessment of visual acuity demonstrated the stabilization of visual acuity and 87.1% (364/418) of eyes were stable, 4.8% (20/418) were improved, and 8.1% (34/418) were aggravated in the long term respectively. The change of visual acuity was frequently seen in the short term, and rapid shifts of body fluid to correct overhydration induced abrupt changes of glycaemic control as well as retraction of macular oedema. Hence it might be one of the factors affecting rapid change of visual acuity in the short term. Meanwhile, long-term deterioration of visual acuity resulted from either hypertension unresponsive to medical treatment or poor glycaemic control. Some DM-ESRD patients had only background retinopathy at the start of haemodialysis and these were likely to have the nephrosclerotic glomerular lesion. They were old, not nephrotic and had a mild degree of diabetes during the predialysis stage. Thus, DM-ESRD patients seem to have some heterogeneity in their clinical characteristics, and old DM-ESRD patients with only background retinopathy have the appearance of diabetic macroangiopathy rather than microangiopathy. PMID- 8414160 TI - Effect of age on protein catabolic rate, morbidity, and mortality in uraemic patients with adequate dialysis. AB - To test the validity of the assumption that the protein catabolic rate (PCRn g/kg/day) is dependent on the normalized dose of dialysis (Kt/V urea), and to try to define the characteristics of the patients in the undefined domain A of the mechanistic map of the National Cooperative Dialysis Study (NCDS), which should include patients with adequate amount of dialysis but inadequate PCRn, urea kinetic modelling was performed over 12 months on 85 patients undergoing haemodialysis. All the patients were managed to maintain a Kt/V urea > or = 0.9. During the entire period of study the total number of hospitalizations and the number of days of hospitalization were recorded. Total serum proteins and serum albumin concentrations were measured at the start and at the end of the study. The results of the study show that there was no correlation between Kt/V and PCRn nor between Kt/V and patient's age, but there was a strong inverse correlation between age and PCRn (r = 0.578; P < 0.0001). Further division of the patients into four groups according to age showed that the lowest values of PCRn were for the group of patients > or = 75 years old. Twelve patients with PCRn < or = 0.8 and Kt/V > or = 0.9 were included in domain A of the mechanistic map. Eleven (92%) of these 12 patients were > or = 65 years old. No correlations were found between the total number of hospitalizations, the total days of hospitalization, Kt/V, time on HD, body weight and PCRn by multiple regression analysis, while the inverse correlation between PCRn and age was confirmed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414161 TI - Effect of cyclosporin A on endothelin synthesis by cultured human renal cortical epithelial cells. AB - We report here for the first time that human renal proximal tubular cells secrete endothelin, clear evidence of de-novo endothelin synthesis by these cells and the effect of cyclosporin A (CsA) on endothelin synthesis both in short-term (24 h) and medium-term (5-day) culture. Human renal cortical epithelial cells were cultured and shown to possess proximal tubular characteristics. These cells produced endothelin in culture in a time-dependent manner, as measured by radioimmunoassay (291.6 +/- 51.4 pg/well/24 h). Furthermore, endothelin production by these cells was significantly decreased by up to 80% by cycloheximide (1051.8 +/- 54.9 pg/mg cell protein/24 h versus 253.2 +/- 12.6 pg/mg cell protein/24 h), showing that these cells actively synthesize endothelin. In short-term culture (24 h), CsA significantly inhibited endothelin synthesis at a medium concentration of 10,000 micrograms/l. No change in endothelin synthesis was seen at lower CsA concentrations. In contrast, over a 5 day period, a non-significant increase in endothelin synthesis was observed at CsA concentrations of 2000 micrograms/l (152.5 +/- 20.4%); however, cell growth was significantly decreased at this concentration (71.33 +/- 6.39%). Using a newly developed two-site immunoradiometric assay specific for endothelin-1 (ET 1), we demonstrate that ET-1 is the major endothelin isoform produced by human renal proximal tubular cells. PMID- 8414162 TI - Pauci-immune glomerulonephritis associated with bacterial infection. PMID- 8414163 TI - Echinococcosis of the kidney. PMID- 8414164 TI - Klatskin's tumour 10 years after successful cadaveric renal transplantation. PMID- 8414165 TI - Mesangiocapillary glomerulonephritis associated with hydatid disease. PMID- 8414166 TI - Relation between serum IGF-I and anaemia in predialysis chronic renal failure patients. PMID- 8414167 TI - Pseudoaneurysm of femoral artery. Therapeutic role of interventional radiology. PMID- 8414168 TI - Recovery of renal function in a patient with haemolytic-uraemic syndrome and severe renal involvement. PMID- 8414169 TI - Minimal-change nephropathy: how does the immune system affect the glomerulus. PMID- 8414170 TI - Preliminary results of a magnetic resonance imaging (MRI) study of the pig brain placed in stereotaxic conditions. AB - Magnetic resonance imaging (MRI) of two pig brains was realized. The animals were placed in the stereotaxic conditions currently used in experiments. To allow the positioning of the animal in the MRI instrument, landmarks were previously traced on the snout of the pig. To avoid movements, animals were deeply anesthetized. MRI were taken in frontal, horizontal and sagittal directions. Afterwards, the brains of the pigs were frozen and cut into sections, frontal for one animal and sagittal for the other. Histologic and MR images were compared. The usefulness of this technique is discussed. PMID- 8414171 TI - Effect of prolactin on the sleep-wake cycle in the rat. AB - The purpose of this study was to examine the effects of prolactin on the sleep wake cycle. Ovine prolactin was injected subcutaneous at a dose of 10 micrograms/animal or intra-cerebro-ventricularly at doses of 100, 10 and 1 ng/animal. The subcutaneous injections were given during either the diurnal or nocturnal period. Results indicate that oPRL decreases paradoxical sleep duration when injected during the dark period and increases it when injected during the light period. The i.c.v. injections were given only during the dark period and the effects were similar to those obtained with the s.c. injections. There was no effect on slow-wave sleep duration irrespective of injection time or injection site. PMID- 8414172 TI - Corticosterone response to the serotonergic agonist D-fenfluramine may be independent from corticotropin-releasing factor (CRF). AB - This study was designed to assess the involvement of corticotropin-releasing factor (CRF) in the corticosterone response to the acute administration of the serotonergic indirect agonist D-fenfluramine in the rat. In addition to plasma corticosterone, D-fenfluramine-induced hyperglycemia (which is independent from the hypothalamo-pituitary-adrenal axis) was also analyzed. Acute i.v. injection of sheep anti-CRF antiserum (15 min beforehand) markedly diminished either stress induced corticosterone release (but not ether stress-induced increases in plasma glucose levels), thereby indicating that passive immunization was efficient. Acute administration of D-fenfluramine (3 mg/kg i.v.) increased plasma corticosterone and glucose levels to similar extents in control rats (i.e. injected with normal sheep serum) and in anti-CRF antiserum-injected rats. These results indicate that, under our experimental conditions, D-fenfluramine-induced corticosterone elevation is of peripheral origin (through pituitary and/or adrenocortical pathways). PMID- 8414173 TI - Ontogeny of gene expression of inositol 1,4,5-trisphosphate receptor in the rat brain: high mRNA levels in the cerebellar Purkinje cells. AB - The release of intracellular Ca, which is involved in many neuronal functions, is regulated by the second messenger inositol 1,4,5-trisphosphate (InsP3) interacting with specific receptor. The distribution of the mRNA coding for the recently cloned InsP3 receptor was studied in the developing rat brain using oligonucleotides derived from the rat cDNA sequence and in situ hybridization. The localizations of the mRNA in the postnatal brain were exactly superimposable to that previously reported in the adult [Mailleux et al., Neuroscience, 49 (1992)577-590]. Higher mRNA levels were consistently found in the adult neurons over their postnatal counterpart. Hybridization signal was first visible in the cerebellar Purkinje cells which express dramatically higher mRNA levels of the receptor than any other neurons in the brain. In conclusion, the levels of InsP3 receptor mRNA per neuron increased with synaptogenesis. This finding suggests the occurrence during this critical developmental period of a more complex regulation of Ca fluxes, perhaps requiring higher intraneuronal levels of InsP3 receptor. PMID- 8414174 TI - Interaction of ethanol with muscarinic receptor-stimulated phosphoinositide metabolism during the brain growth spurt in the rat: role of acetaldehyde. AB - The developing brain is extremely sensitive to the neurotoxicity of ethanol; however, the mechanism(s) of its developmental neurotoxicity are still elusive. In the developing rat brain, ethanol exerts an age-, brain region-, and receptor specific inhibitory effect on muscarinic receptor-stimulated phosphoinositide metabolism, which may be linked to some of the neurotoxic effects of ethanol found in children with fetal alcohol syndrome. Since some studies have suggested that the ethanol metabolite acetaldehyde may mediate, at least in part, the developmental effects of ethanol, in the present study we have examined whether acetaldehyde would inhibit carbachol-stimulated phosphoinositide metabolism in brain slices from immature rats. We also tested propionaldehyde, the corresponding aldehyde of n-propanol, another alcohol shown to cause microencephaly and to affect phosphoinositide metabolism in the developing rat. Neither acetaldehyde nor propionaldehyde, at concentrations up to 1 mM, had any inhibitory effect on this system, while the two alcohols did, as previously reported. These results suggest that ethanol itself may be the primary agent responsible for its developmental neurotoxicity. PMID- 8414175 TI - Identification and functional significance of nicotinic cholinergic receptors in the rat pineal gland. AB - The existence and subunit identification of the nicotinic cholinergic receptors in the rat pineal gland were examined by autoradiography, using [3H]cytisine and [125I]alpha-bungarotoxin as labelled ligands. The experiments performed with radioactive cytisine did not reveal specific binding, while iodinated alpha bungarotoxin disclosed moderate specific binding density, suggesting that the nicotinic cholinergic receptor in the rat pineal is structurally organized with the alpha 7 or alpha 8 subunits present, the only ones that bind alpha bungarotoxin with high affinity. In vitro functional experiments using pineal explants demonstrated that the binding site may represent a readily accessible nicotinic cholinergic receptor. Nicotine, though having no effect per se on the synthesis and release of melatonin, significantly diminished, in a dose-dependent manner, the norepinephrine-stimulated melatonin accumulation. This effect could be blocked by coincubation with the cholinergic antagonist d-tubocurarine, suggesting that the nicotinic cholinergic receptor in the rat pineal could be involved in the functional regulation of the gland. PMID- 8414176 TI - Inhibition of volume regulation and efflux of osmoregulatory amino acids by blockers of Cl- transport in cultured astrocytes. AB - Regulatory volume decrease (RVD) in astrocytes was inhibited by the Cl- exchanger blockers 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), dipyridamole and niflumic acid but not by the Cl- channel inhibitors diphenylamine-2 carboxylate (DPC) and anthracene-9-carboxylate (9-AC). The volume activated efflux of [3H]taurine and [3H]D-aspartate (as marker for glutamate) was similarly affected by these compounds. However, neither RVD nor osmolyte fluxes were significantly reduced by removal of external Cl-, suggesting that an anion exchanger activity is not required for the volume regulatory process. Alternatively, these results suggest that the anion exchanger molecule may function as an unidirectional Cl- channel possibly permeable also to amino acids. PMID- 8414177 TI - Mossy fiber sprouting in epileptic rats is associated with a transient increased expression of alpha-tubulin. AB - Kainate-induced seizures lead to marked increases of alpha-tubulin mRNA and protein immunoreactivity in the rat dentate gyrus. The increase in alpha-tubulin mRNA was restricted to the granule cell bodies. alpha-Tubulin immunoreactivity was enhanced in granule cell dendrites and axons (the mossy fibers), in the molecular layer. These changes peaked 6-12 days after kainate treatment and preceded the collateral sprouting of mossy fibers which occur 12 to 30 days after seizures. The present results suggest that microtubule formation contributes to the synaptic rearrangements which take place in the hippocampus after seizures. PMID- 8414178 TI - Glutathione modulates the N-methyl-D-aspartate receptor-activated calcium influx into cultured rat cerebellar granule cells. AB - The effects of reduced (GSH) and oxidized (GSSG) glutathione and dithiothreitol (DTT) and L-cysteine on the influx of 45Ca2+ were studied with cultured cerebellar granule cells. DTT slightly enhanced the basal influx but strongly activated the influx stimulated by glutamate or N-methyl-D-aspartate (NMDA). The effects on the kainate- or quisqualate-induced influx were less pronounced. Extracellular GSH had no effect on the basal influx of Ca2+. A concentration of 0.5 mM GSH slightly activated the glutamate- and NMDA-induced influx while GSSG was inhibitory. The enhancement by DTT and cysteine of the responses to excitatory amino acids was attenuated by GSH and GSSG. We propose that both the accessibility and redox state of the functional sulfhydryl groups in NMDA receptor-ionophores may be regulated by endogenous glutathione. These effects are attributed to the gamma-glutamyl moiety and sulfhydryl group in the tripeptide molecule. PMID- 8414179 TI - Hippocampal interneuron activity in unanesthetized rats: relationship to the sleep-wake cycle. AB - Evoked population spikes and interneuronal discharges were recorded throughout the sleep-wake cycle in hippocampal regions CA1 and dentate gyrus (DG) of ten chronically implanted rats. During quiet wakefulness (QW) and slow-wave-sleep (SWS) (non-theta rhythm states), the primary shock of paired stimuli evoked in CA1 both high amplitude population spikes and multiple interneuron discharges when compared to active wakefulness (AW) and rapid-eye-movement (REM) sleep (theta rhythm states). A second shock was delivered to CA1 afferents 60 ms after the first shock. This second shock evoked a small population spikes during non theta states, whereas it evoked higher amplitude population spikes in theta states. The second shock also evoked unit interneuron discharges in non-theta states but not in theta states. In the dentate gyrus, identical primary afferent stimulation evoked similar interneuron activity and uniform amplitude population spikes throughout the sleep-wake cycle. In contrast, the secondary shocks evoked a striking potentiation of the field population spike during sleep, SWS and REM sleep compared to AW and OW. Evoked DG interneuron spikes following the second population spike were greater in number during SWS compared to the other stages. Our findings suggest that hippocampal field potentials and interneuron activity recorded in vivo are regionally regulated, have unique state-dependent expression and are strongly influenced by inhibitory feed-forward mechanisms. PMID- 8414180 TI - Differential expression of GABA transporter-1 messenger RNA in subpopulations of GABA neurones. AB - Using in situ hybridisation with an oligonucleotide probe, the regional distribution of GABA transporter-1 messenger RNA was determined in the adult rat CNS. Overall, the distribution of GABA transporter-1 mRNA was similar to that of glutamate decarboxylase-67 mRNA, consistent with an expression in GABA neurones. However, in cerebellar cortex, Purkinje cells which express high levels of glutamate decarboxylase-67 mRNA did not express GABA transporter-1 mRNA, whereas Bergmann glia expressed GABA transporter-1 mRNA at high levels. In some other brain regions, including the inferior colliculus and reticular thalamus, there was no GABA transporter-1, although there were neurones which expressed glutamate decarboxylase-67 messenger RNA at high levels. PMID- 8414181 TI - Spinal and supraspinal mechanisms contribute to the silent period in the contracting soleus muscle after transcranial magnetic stimulation of human motor cortex. AB - In the voluntarily activated muscle, transcranial magnetic stimulation (TMS) of motor cortex produces subsequently to the motor evoked potential (MEP) a silent period (SP) in the electromyogram. We studied the time course of soleus motoneuron (MN) pool excitability after conditioning TMS by Hoffmann reflex (HR) testing, to determine whether inaccessibility of MNs after corticospinal input contributes to the SP. Coincidently with the early part of the SP, and only in the contracting soleus, MN depression was obtained that covaried with the degree of preinnervation, and with the size of the preceding MN discharge. However, MN excitability recovered significantly prior to the end of the SP. It is concluded that in the contracting soleus spinal mechanisms (most likely Renshaw inhibition and MN afterhyperpolarization) contribute to the early part of the SP, while the late part of the SP is supraspinal (probably cortical) in origin. PMID- 8414182 TI - Increased loss of brain DNA in the neonatal vasopressin-deficient Brattleboro rat, but not in normal rat treated with vasopressin antagonist. AB - In order to establish whether vasopressin (VP) influences brain cell survival, [3H]thymidine was injected in 10-day-old vasopressin-deficient Brattleboro rat pups, as well as in Wistar pups treated, neonatally, with the VP antagonist dP[Tyr(Me)2]VP followed by subsequent measurement of [3H]DNA in olfactory bulbs and cerebellum days and weeks thereafter. Results show, first of all, that the incorporation of [3H]thymidine into DNA was enhanced in the homozygous (HOM) Brattleboro, when compared with the heterozygous (HET; non-vasopressin-deficient) controls. The difference is due to the greater and prolonged tissue availability of [3H]thymidine, possibly pointing to an altered thymidine uptake and/or metabolism. Between postnatal days 25 and 39 no differences were seen in [3H]DNA content of the brain parts of the HET and Wistar control rats. For the HOM rats, however, a loss of [3H]DNA was seen (up to 8%), indicating that increased postnatal brain cell death might occur in the mutant. The antagonist treatment in Wistar rat up to 21 days of age failed to show a similar effect. It is proposed that general growth impairments, rather than VP receptor-mediated effects, lead to the brain cell loss. PMID- 8414183 TI - Factors in the cerebrospinal fluid of multiple sclerosis patients interfering with voltage-dependent sodium channels. AB - The effect of cerebrospinal fluid (CSF) from patients with multiple sclerosis (MS) on voltage-dependent Na+ channels in human myoballs was studied. The transient Na+ currents, elicited by whole-cell depolarization from -85 to -20 mV, were decreased to 75-25% the control value in the presence of CSF from all 7 MS patients investigated. The effect was complete in about 5 s and was fully reversible on admission of standard external fluid. Such decrease was not or only to a minor extent observed with 10 out of 11 control CSFs from patients without inflammatory neurological disease. The origin of the factors interfering with the Na+ channels is unknown. It is suggested that, in addition to demyelination, impaired Na+ channel function might cause the symptoms in MS. PMID- 8414184 TI - Immunohistochemical demonstration of protein gene product 9.5 (PGP 9.5) in the primary olfactory system of the rat. AB - Immunohistochemical localization of protein gene product 9.5 (PGP 9.5) was examined in the primary olfactory system of the rat. Receptor cells were intensely immunopositive in the olfactory epithelium, vomeronasal organ and septal olfactory organ of Masara. The mitral cells and some of the short-axon cells of the main olfactory bulb (MOB) were also intensely immunopositive, while the tufted cells of the MOB and the mitral/tufted cells of the accessory olfactory bulb varied in immunoreactivity. PMID- 8414185 TI - Ethanol inhibits tetraethylammonium chloride-induced synaptic plasticity in area CA1 of rat hippocampus. AB - Dendritic recordings of the rat hippocampal formation were used to assess the ability of ethanol to inhibit the induction of synaptic plasticity due to bath application of the K+ channel blocker, tetraethylammonium chloride (TEA). Brief application of TEA resulted in the enhancement of the population excitatory postsynaptic potential (EPSP) slope by an average of 26 +/- 3% over the pre-TEA baseline. However, coapplication of TEA and ethanol (100 mM) resulted in enhancement of only 10 +/- 2% of the EPSP slope. These data suggest that LTP (long-term potentiation) due to the activation of voltage-gated Ca2+ channels is also inhibited by ethanol. PMID- 8414186 TI - The NMDA receptor antagonist MK-801 markedly reduces the induction of c-fos gene by haloperidol in the mouse striatum. AB - The influence of the N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 (dizocilpine), on the haloperidol-induced increase of c-fos mRNA levels in the striata of mice was examined using the quantitative Northern blot analysis method. Administration of haloperidol (1 mg/kg. i.p.) increased striatal c-fos mRNA hybridization signal about 7-fold, as measured 30 min after injection. MK 801 (0.2-4.5 mg/kg, i.p.) dose-dependently reduced this effect of the dopamine antagonist. Higher doses of MK-801 (1.5-4.5 mg/kg) diminished c-fos mRNA increase produced by haloperidol by 80%. Such a marked inhibition of this haloperidol effect by MK-801 has not been previously reported. It suggests that the induction of c-fos gene occurring in the striatum after dopamine D2 receptor blockade is mediated in great part by endogenous glutamate acting at NMDA receptors. Since the c-fos gene protein product (Fos) is a transcription factor, our results also indicate the involvement of glutamate in the regulation of expression of other genes in the striatum. PMID- 8414187 TI - Sustained hyperalgesia can be induced in the rat by a single formalin injection and depends on the initial nociceptive inputs. AB - The present study was undertaken to find out if a hyperalgesia can be observed 1 week after a conditioning stimulation of the orofacial area of the rat. Sprague Dawley rats received a left infraorbital infiltration with either bupivacaine or saline and then, 30 min thereafter, an injection of either saline or 10% formalin solution in the left upper lip. Four groups of animals were thus made up depending on their conditioning treatment. Seven days later, an algesimetric test initiated by a contralateral orofacial formalin injection was carried out. The duration of lip rubbing was significantly increased in formalin-conditioned groups. The hyperalgesia observed at 7 days was suppressed by an infraorbital nerve block carried out at the time of the conditioning stimulus. These results tend to indicate that a nociceptive message of short duration induces a trace in the central nervous system which can be retained for 1 week. PMID- 8414188 TI - Effect of adrenal demedullation on neuropeptide Y content of the capsule/glomerulosa zone of the rat adrenal gland. AB - The capsule/glomerulosa zone of the adrenal gland is richly innervated by neuropeptide Y (NPY)-containing nerve fibers. The content and concentration of NPY in the capsule/glomerulosa zone of the female rat adrenal were determined by radioimmunoassay both in unoperated females (controls) and in operated ones, a week after in situ unilateral demedullation which excludes both the medulla and the fasciculata/reticularis zones. Demedullation induced a significant weight increase of the capsule/glomerulosa zone of the operated gland (compared to contralateral intact one) as well as of the medulla/fasciculata-reticularis zone of the contralateral intact one (compared to the corresponding part of the gland of controls). Both NPY content and concentration in the capsule/glomerulosa zone of the demedullated adrenal were significantly reduced in comparison with those in the corresponding part of the contralateral intact gland. NPY immunoreactive fibers, revealed by immunofluorescence, were present in the capsule/glomerular zone of both intact and contralateral demedullated adrenal gland. In the former, NPY fibers were regularly distributed in this part of the gland, while in the latter, some areas were devoid of immunoreactive fibers. NPY content, but not concentration, was increased in the medulla/fasciculata-reticularis zone of the contralateral intact gland. Present data support a dual origin for the NPY nerves present in the capsule/glomerulosa zone of the adrenal cortex: one part could arise from extra-adrenal site possibly the suprarenal ganglia while the other part could arise from intra adrenal ganglia cells which also contain NPY. PMID- 8414189 TI - Glucocorticoid regulation of cannabinoid receptor messenger RNA levels in the rat caudate-putamen. An in situ hybridization study. AB - The influence of adrenalectomy on cannabinoid receptor gene expression in the adult rat dorsal striatum was investigated by in situ hybridization with specific oligonucleotide probes. The caudate-putamen of adrenalectomized rats contains approximately 50% significantly higher mRNA levels than the controls. This increase could be counteracted by dexamethasone treatment. Together, these results suggest glucocorticoid down-regulation of cannabinoid receptor gene expression in the striatum. PMID- 8414190 TI - VIP-induced stellation and immediate early gene expression in astrocytes: effects of dexamethasone. AB - To investigate the actions of glucocorticoids (GCs) on astrocyte functions, interactions of dexamethasone and immediate early genes (IEGs) were studied in cell cultures of rat cerebral cortical astrocytes. Vasoactive intestinal peptide (VIP) induces rapid c-fos mRNA expression and morphological changes (stellation) in cultured astrocytes. Dexamethasone pretreatment decreases this ligand-induced stellation without affecting levels of c-fos mRNA. Moreover VIP does not induce c jun, jun-B, and NGFI-A mRNA, suggesting that these IEGs may not mediate ligand induced stellation. The expression of c-fos, c-jun, jun-B, and NGFI-A mRNA are rapidly induced in cultured astrocytes after treatment with phorbol ester, epidermal growth factor, and basic fibroblast growth factor. Dexamethasone pretreatment has no effect on the IEG response induced by any of these agents, suggesting that GCs may not have direct effects on the promoter of these IEGs in cortical astrocytes. PMID- 8414191 TI - Okadaic acid enhances abnormal phosphorylation on tau proteins. AB - Tau proteins are one of the microtubule-associated proteins (MAPs) and show promoting activity on microtubule assembly. Tau proves to be the major constituent of Alzheimer's paired helical filaments, in which tau is found to be different from normal tau in that it is abnormally phosphorylated. To examine the effect of the abnormal phosphorylation on microtubule assembly, we obtained abnormally phosphorylated tau that was made in vitro by hyperphosphorylation with ATP or with ATP and okadaic acid, a drug inhibiting phosphatase, mainly 1 and 2A. We confirmed the biochemical properties of abnormally phosphorylated tau based on its retarded gel mobility and immunoreactivity to anti-PHF. We found that abnormally phosphorylated tau was able to promote the polymerization of microtubules but showed less activity as compared with normally phosphorylated tau. This effect of ATP on abnormal phosphorylation of tau was enhanced when okadaic acid was added in the phosphorylation reaction mixture during microtubule assembly. It is of significance that phosphatase activity as well as kinase activity are involved in the formation of abnormal tau. The present evidence suggests the simultaneous occurrence of microtubule disassembly and the pathogenesis of paired helical filaments following the abnormal phosphorylation of tau. PMID- 8414192 TI - Vesicle monoamine transporters 1 and 2: differential distribution and regulation of their mRNAs in chromaffin and ganglion cells of rat adrenal medulla. AB - The expression and synthesis regulation of the vesicle monoamine transporter was investigated in rat adrenal medulla. Previous studies established two genes for monoamine transporters by molecular techniques. In rat adrenal medulla, a differential expression of the corresponding mRNAs was found by in situ hybridization. The mRNA of monoamine transporter 2 was localized in chromaffin cells whereas monoamine transporter 1 mRNA occurred only in ganglion cells of the adrenal medulla. Insulin-induced hypoglycemia, a model for short neurogenic stimulation of the adrenal medulla, did not alter steady-state mRNA levels of both monoamine transporters. PMID- 8414193 TI - NMDA receptors at excitatory synapses in the hippocampus: test of a theory of magnesium block. AB - Excitatory postsynaptic currents (EPSCs) were studied using the whole-cell patch clamp technique to record from rat hippocampal pyramidal neurons in both slices and dissociated cultures. The voltage- and magnesium-dependence of the N-methyl-D aspartate (NMDA) component of the EPSCs was quantified by measuring the amplitude of this component at a range of holding potentials (-80 mV to 60 or 80 mV) in several bath magnesium concentrations (0.2 microM, 0.1, 1 and 10 mM). All of our results were well-fitted by a theory of magnesium block developed from single channel studies of extrajunctional NMDA channels in culture. It is concluded that extrasynaptic and subsynaptic NMDA channels both in culture and in situ have identical properties of magnesium block. PMID- 8414194 TI - Barbiturate anaesthesia reduces the neurotoxic effects of quinolinate but not ibotenate in the rat pedunculopontine tegmental nucleus. AB - We have previously demonstrated that ibotenate (IBO) injected into the pedunculopontine tegmental nucleus (PPTg) damages all neurones there while quinolinate (QUIN) makes relatively selective lesions of cholinergic neurones. We now compare the effects of two anaesthetics, sodium pentobarbitone and Avertin (tribromoethanol/tert-amylalcohol dissolved in ethanol, saline and phosphate buffer) on three doses of IBO and QUIN in the PPTg. Diaphorase-positive cell loss after QUIN was attenuated under barbiturate, the relative selectivity of QUIN for diaphorase-positive neurones was lost and lesion volumes were uniformly small compared with lesions made under Avertin anaesthesia. IBO toxicity was unaffected by anaesthesia. These data are discussed with reference to the actions of excitotoxins at glutamate receptor subtypes and interactions of barbiturates with the GABAA receptor. PMID- 8414195 TI - Proline-directed kinase systems in Alzheimer's disease pathology. AB - Immunohistochemical analysis was used to assess the distribution of the proline directed kinase, cdc2, in Alzheimer's disease (AD) pathology. A robust signal was most prominent in the neurofibrillary tangle (NFT) of affected neurons that also contained abnormally phosphorylated tau protein. Biochemical analysis identified a pool of cdc2 in bovine brain microtubules that contain normal tau. These results strongly support the hypothesis that cdc2 is involved in the abnormal phosphorylation of tau in AD pathology and they raise important issues regarding regulation of tau phosphorylation in normal and diseased neurons. PMID- 8414196 TI - Expression of smg p25A/rab 3A guanine nucleotide dissociation inhibitor (GDI) in neurons and glial cells from rat brain. AB - smg p25A/rab 3A is a member of the rab family of small guanosine 5'-trisphosphate (GTP)-binding proteins, which are postulated to act as specific regulators of membrane trafficking in exocytosis and endocytosis. In neural tissues, smg p25A/rab 3A is expressed only in neurons but not in glia. In this study, expression of a regulatory protein for smg p25A/rab 3A, guanine nucleotide dissociation inhibitor (smg p25A/rab 3A GDI), in neurons and glia isolated from rat brain was examined. The smg p25A/rab 3A GDI mRNa was detected as early as on gestational day 14 in the fetal brain, and its level reached a maximum 5 days after birth. By immunoblot analysis, smg p25A/rab 3A GDI was detected in the homogenate of rat hippocampus as well as in that of astrocytes in primary culture. By immunocytochemical analysis, the smg p25A/rab 3A GDI immunoreactivity was detected in hippocampal neurons and oligodendrocytes. The immunoreactivity of astrocytes was much lower than that of neurons and oligodendrocytes. These results indicate that smg p25A/rab 3A GDI may play a fundamental role in membrane trafficking both neurons and glia by interacting with multiple rab family proteins. PMID- 8414197 TI - The ventral pallidum area is involved in the acquisition but not expression of the amphetamine conditioned place preference. AB - The present study examined the roles of the ventral pallidum area, one output system of the nucleus accumbens, in the acquisition and expression of the amphetamine conditioned place preference (CPP). Pre-conditioning NMDA lesions of the ventral pallidum area completely abolished the acquisition of the CPP. By contrast, post-conditioning NMDA lesions of the same area had no effect on the expression of the CPP. These results suggest that the ventral pallidum area mediates some process that involves the primary, but not conditioned rewarding effects of amphetamine. PMID- 8414198 TI - Ubiquitin-reactive neurites in cerebral cortex of subjects with Huntington's chorea: a pathological correlate of dementia? AB - We studied the prevalence of ubiquitin-reactive dystrophic neurites in neocortex of cases with Huntington's disease (HD), with a history of dementia lasting from 5 to 8 years before death, and in four age-matched controls. The ubiquitin reactive neurites, identified as round structures localized outside neuronal and glial cells, were quantified in six microscopic fields of cingulate, 2nd temporal and 2nd parietal cortical gyri. The number of ubiquitin-reactive neurites in HD was 12 to 16 times that of controls in the three cortical areas examined. The finding indicates that in HD neocortex there is a severe degeneration of neuronal processes and suggests that it may represent a pathological correlate of dementia. PMID- 8414199 TI - 2-Deoxyglucose autoradiographic evidence that fetal substantia nigra grafts exert contralateral effects in 6-hydroxydopamine-lesioned animals who recover motor asymmetries. AB - Fetal substantia nigra grafts decrease turning behavior induced by dopaminergic agonists in rats with unilateral 6-hydroxydopamine lesions of the nigrostriatal system. The purpose of this study was to determine, through the [14C]2 Deoxyglucose autoradiographic method, whether substantia nigra grafts have an effect on efferent striatal nuclei and other related structures. The results showed an unexpected enhancement of metabolic activity in the substantia nigra pars reticulata and striatum on the side contralateral to the lesion and a bilateral enhancement in the thalamus and frontoparietal cortex, in rats bearing fetal substantia nigra grafts. The results are discussed in terms of the possibility that recovery of turning behavior may in part be caused by an increased activity of the striatum contralateral to the grafted side. PMID- 8414200 TI - Pancreatic pseudocyst infected with Candida albicans. AB - We present a case of a pancreatic pseudocyst secondarily infected with Candida albicans. The authors discuss the risk factors for pseudocyst infection, especially with Candida, diagnostic procedures, and current management including percutaneous catheter drainage. PMID- 8414201 TI - Treatment of upper airway obstruction associated with goiter. AB - The authors examined the resolution of spirometric indices of upper airway obstruction (UAO) following surgical treatment for goiter. The results of a recent study suggest that the integrity of the upper airway remains intact but prolonged recovery is required. PMID- 8414202 TI - Reductionism and encephalitis lethargica, 1916-1939. AB - This paper is the 1992 winner of the Annual Stephen Wickes Prize in Medicine, presented by the Medical History Society of New Jersey. The history of encephalitis lethargica reflects a complex interaction between research-oriented science and clinical medicine. PMID- 8414203 TI - Evaluation and treatment of post-traumatic stress disorder. AB - Post-traumatic stress disorder (PTSD) is a common anxiety disorder seen by general practice physicians as well as by specialists. The authors review current assessment criteria, psychotherapy procedures, and psychopharmacological management of PTSD patients. PMID- 8414204 TI - Clinical imaging rounds: hypertension in a postpartum patient. AB - The authors report a case of a 26-year-old female with gestational headaches, hypertension, weight loss, and tachycardia. Symptoms continued postpartum and a hypertensive workup revealed an adrenal mass and elevated catecholamines. PMID- 8414205 TI - Advance directives for HIV-positive patients. AB - The authors surveyed HIV-positive patients to determine their knowledge of and interest in advance directives and their preferences for end-of-life care. The results of this study make clear the need to institute a program of education for those desiring to know more about advance directives. PMID- 8414206 TI - Salmonella abscess in an ovarian endometrioma. AB - The authors present a patient with an abscess due to Salmonella in an ovarian endometrioma. The apparent inoculation occurred nine months before surgery. This patient had no signs of active infection; her symptoms suggested endometriosis only. PMID- 8414207 TI - Case report: possible Lyme endocarditis. AB - A 56-year-old male presented with clinical and serological data suggestive of Lyme disease. The patient was found to have an aortic valvular vegetation by transesophageal echocardiography. This report represents the first case of Lyme disease with possible valvular involvement. PMID- 8414208 TI - Laboratory improvement: the dirty dozen for OSHA. AB - Safety practices are a necessary part of the physician office laboratory operation. The Occupational Safety and Health Administration (OSHA) inspects office laboratories constantly and a physician must be aware of the condition of the laboratory at all times. PMID- 8414209 TI - Crisis intervention on a house call: a family doctor's way. AB - Having made over 50,000 house calls during his lengthy career, the author writes of a special house call. The author uses this house call as a learning experience by relating the events that took place during the house call. The lesson to learn is that the physician can heal a patient in many ways. PMID- 8414210 TI - Chondrosarcoma of the cricoid: the role of larynx preservation. AB - Chondrosarcoma of the cricoid cartilage historically has been treated by total laryngectomy. Recent reports lend support for the role of local excision. We present one case and discuss the surgical management for a 63-year-old patient with excellent results. PMID- 8414211 TI - Imaging of herpes simplex encephalitis. AB - The authors discuss computed tomography (CT) and magnetic resonance imaging (MRI) findings of herpes encephalitis as important adjuncts to the early diagnosis of this entity. The patient was a 42-year-old HIV-negative male who presented with a history of dizziness, headaches, and fever. PMID- 8414212 TI - An intensivist's view: who should be admitted to the intensive care unit? Society of critical care. PMID- 8414213 TI - Extracellular matrix proteins with neurite promoting activity and their receptors. AB - Characteristic features of the nervous system converge into network formation during the development. The neurons recognize precisely their target cells and form synapses, and these steps are complex, but well organized spatially and temporally. The neurite promotion from the neurons is one of the most important events for synapse formation. It is well known that extracellular matrix proteins such as laminin and their receptors, and cell adhesion molecules such as NCAM participate in cell migration and synaptic formation. We have isolated a neurite outgrowth factor (NOF) which promotes neurite outgrowth from various neurons and belongs to laminin family, and also its receptor which is identified to be an immunoglobulin superfamily protein by cDNA cloning. This ligand-receptor system is a unique example that a receptor with immunoglobulin-like structure interacts with an extracellular matrix protein. PMID- 8414214 TI - A cerebral ischemia model produced by injection of microspheres via the external carotid artery in freely moving rats. AB - We produced an improved microembolism model of cerebral focal ischemia by injection of 1000-2000 microspheres (50 +/- 5 microns diameter) via a tube retrogradely inserted into the right external carotid artery in freely moving rats. The group injected with 2000 spheres showed a much more severe mortality rate as well as neurological signs than did the 1000-sphere group. Brain water content of the 2000-sphere group was examined and found to show an increase from 4 to 24 h after embolization in the right hemisphere, indicating serious brain edema. Severe neurological signs and individual deaths by embolization were most likely related to the extent of development of brain edema. Examination of learning behavior by shuttle-box avoidance revealed partial but significant impairment of learning in the 1000-sphere group. Autoradiographic studies for muscarinic acetylcholine receptors and protein kinase C binding sites were conducted. Both these binding sites decreased in number, but protein kinase C seems to be more susceptible to ischemic injury than muscarinic acetylcholine receptors. The observation was considered to be closely related with an impairment of learning. The present study suggests that our microembolism model in freely moving rats is useful for investigations of the early phase and late phase of cerebral ischemia. PMID- 8414215 TI - Immobilization stress induces alterations of second-messenger systems in the gerbil brain. AB - The effects of immobilization stress on the cerebral second messenger (adenylate cyclase and phosphoinositide) were investigated autoradiographically in mongolian gerbils. After 10 min (10-min stress group, n = 7), or after 6 h (6-h stress group, n = 7) of fixation on a flat board while supine, in vitro autoradiography was performed using [3H]forskolin (3H-FK) and [3H]phorbol-12,13-dibutyrate (3H PDBu) as specific ligands to identify the distribution of adenylate cyclase and protein kinase C, respectively. In another group of 7 gerbils (control group), the same autoradiographic procedure was performed immediately after the animals were removed from the cage. In the 10-min stress group, FK binding was significantly decreased in the hypothalamus and amygdala, but significantly increased in the basal ganglia including the caudate-putamen and globus pallidus. FK binding in the 6-h stress group tended to increase throughout the brain, rising significantly in the basal ganglia. PDBu binding in either stress group did not change significantly compared to the control group in any region except the hippocampal CA3 region of the 6-h stress group. Under immobilization stress, the adenylate cyclase system may undergo time-dependent and regionally specific changes, while the phosphoinositide system remains relatively stable. PMID- 8414216 TI - Cortical cell types from spike trains. AB - The patterns of cell activity recorded extracellularly in the motor cortex of behaving monkeys were classified into the following three groups using a combination of cluster and discriminant analyses of 1925 spike trains: (a) cells with low discharge rate and low bursting (67.1%), (b) cells with low discharge rate but bursting (20.2%), and (c) cells with high discharge rate and low bursting (12.7%). The percentage of directionally tuned cells and of cells engaged during a memorized delay task were very similar in all three cell groups. PMID- 8414217 TI - Transient expression of calretinin during development of chick cerebellum. Comparison with calbindin-D28k. AB - Calcium ions play a critical role in neural development. Insights into the ontogeny of Ca2+ homeostasis were gained by investigating the developmental expression of two E-F hand calcium-binding proteins. Calretinin and calbindin were monitored through their immunoreactivity in the developing chick cerebellum (from E6 to E20). Calbindin was detected from E13 and in Purkinje cells only. Intensity of labelling increased with Purkinje cell development. Calretinin presented a transitory immunoreactivity between E11 and E20 in the internal granular cell layer. This cell layer contains cells which will differentiate into Golgi and granular cells which are calretinin-negative in adult chick cerebellum. Calretinin immunoreactivity presented a peak (both in number of cells and in intensity) at E15 and fell dramatically after E20 while calbindin immunoreactivity was restricted to the Purkinje cells and increased with the development of these cells. PMID- 8414218 TI - Topographic relationship between anteromedial thalamic nucleus neurons and their cortical terminal fields in the rat. AB - The present study has examined the topographic relationship between cells in the anteromedial thalamic nucleus (AM) and their cortical terminal fields, with retrograde transport of Fluoro Gold in the rat. Projections to the frontal area 2 originate from the ventrolateral part of the AM and the entire interanteromedial nucleus (IAM). Projections to the anterior cingulate area originate from the peripheral part of the rostral AM and the entire IAM. Fibers to the rostral retrosplenial area arise from the caudodorsal part of the AM, whereas those to the caudal retrosplenial area arise from the rostralmost and the rostrodorsomedial parts. Fibers to the rostral area 29D originate from the rostrocentral part of the AM, whereas those to the caudal area 29D originate from the rostroventrolateral and the ventromedial parts. Projections to the medial half of the entorhinal area originate from the rostrodorsomedial part of the AM. In contrast, projections to the lateral half of the entorhinal area originate from the IAM and the central part of the AM. The results show a complex topographic relationship between cells of origin of the AM and their cortical terminal fields, suggesting complex functional roles played by the AM in learning behavior such as discriminative avoidance behavior. PMID- 8414219 TI - Regulation of nerve growth factor secretion in L-M cells by catechol derivatives. AB - We investigated the mechanism responsible for the stimulation of nerve growth factor (NGF) secretion by catechol derivatives in L-M cells, using L-threo-3,4 dihydroxyphenylserine (L-DOPS). Treatment of the cells with L-DOPS increased the NGF content in the L-M cell medium by approximately 3-fold. This stimulatory effect was not blocked by a decarboxylase inhibitor, or by alpha- or beta adrenergic blockers. Intracellular cAMP levels were not changed by exposure to L DOPS. The antioxidants, ascorbic acid and sodium pyrosulfite, completely prevented the stimulatory effect of L-DOPS, and radical scavengers (superoxide dismutase plus catalase) caused a significant partial inhibition of the response to L-DOPS. Quinone derivatives (adrenochrome, 4-n-propyl-1,2-benzoquinone), which are the oxidative products of the catechol derivatives, increased the NGF content in the medium, and their potency was greater than that of the catechol derivatives themselves. These findings suggest that L-DOPS and other catechol derivatives might be oxidized in the medium to form quinone derivatives, and that it is these which predominantly express a stimulatory effect on NGF secretion by a novel cAMP-independent mechanism in L-M cells. PMID- 8414220 TI - Activities of the intralaryngeal muscles during electrically induced vocalization in decerebrate cats. AB - Electromyograms were recorded from the intralaryngeal muscles (the posterior cricoarytenoid (PCA) and thyroarytenoid (TA) muscles) and the ventilatory muscles (the diaphragm (DA) and abdominal (RA) muscle) during electrically induced vocalization in precollicular postmammillary decerebrate cats. Electrical stimulation (rectangular pulses, 0.2 ms, 10-60 microA, 100 Hz, lasting 5-10 s) delivered to the rostral pons (Horsley-Clarke coordinates, A 1.0 to P 2.0, L or R 3.0 to 5.0, H -4.5 to -6.0) induced alternate inspiration and vocalization. The normal respiratory rhythm was reset to a new one and the intralaryngeal and ventilatory muscles were coordinately activated to produce vocalization. During the period of the electrical stimulation, preceding activity of PCA to DA and postinspiratory activity of DA, which were normally observed during quiet breathing, disappeared. These results suggested that the tonic electrical stimulation delivered to the rostral pons acted as a command signal to alter teh pattern generation of the ventilatory and laryngeal systems from quiet breathing to vocalization. PMID- 8414221 TI - Termination mode and branching patterns of reticuloreticular and reticulospinal fibers of the nucleus reticularis pontis oralis in the cat: an anterograde PHA-L tracing study. AB - By utilizing an anterograde neural tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), pontomedullary reticuloreticular connections and reticulospinal connections were studied, including their fiber trajectories and distribution of PHA-L labeled terminals in close apposition to target reticular and spinal neurons, and branching patterns of axon collaterals at the levels of the cervical and upper thoracic cord. PHA-L was focally microinjected into the medial pontine reticular formation corresponding to the nucleus reticularis pontis oralis. A great number of PHA-L labeled thin fibers descended bilaterally coursing through the medial part of the pontine and medullary reticular formation with an ipsilateral predominance. Labeled terminal boutons were closely apposed to somata of various sized pontomedullary reticular neurons. Labeled thick fibers descended ipsilaterally coursing through the ventral half of the medial longitudinal fasciculus, and further descended through the ventral funiculus of the spinal cord. At the levels of the cervical and upper thoracic cord, these reticulospinal fibers gave off axon collaterals sending terminal fibers to small- to large-sized neurons in Rexed's laminae VII and VIII. Some of the axon collaterals innervated not only ipsilateral but also contralateral gray matter. By reconstructing branching patterns of axon collaterals, each axon collateral was found to innervate spinal neurons located in a disk-like spinal segment with a width less than 1 mm. PMID- 8414222 TI - Vitamin B6 and cognitive development: recent research findings from human and animal studies. AB - Numerous studies have suggested that pregnant and lactating women may have dietary intakes of vitamin B6 that are well below the recommended dietary allowance, which may affect the vitamin B6 status of their offspring. This nutrient is an essential cofactor in the developing central nervous system and may influence brain development and cognitive function. Recent work in animal models suggests that vitamin B6 deficiency during gestation and lactation alters the function of N-methyl-D-aspartate receptors, a subtype of receptors of the glutamatergic neurotransmitter system thought to play an important role in learning and memory. PMID- 8414223 TI - Plant sources of provitamin A and human nutriture. AB - Hypovitaminosis A is a problem in many parts of the developing world. Beyond the stop-gap measures of capsule distribution and food fortification, increased consumption of accessible sources of vitamin A, specifically of the carotenoid provitamin A in yellow, orange, and green plants, has been promoted as the sustainable, long-term solution. However, a search of the available literature reveals few examples of human studies to support the effectiveness of this solution. Evidence from feeding studies shows an almost universally poorer uptake of intact carotenoids from plant sources as opposed to pure, chemical sources. With notable exceptions, the bioconversion of plant carotenoids to preformed vitamin A also seems to be inefficient. Epidemiologic observations in poor Third World populations and in vegetarians in an industrialized nation indicate a relatively greater potency for animal sources of vitamin A. In developing countries, low fat intakes, intestinal roundworms, recurrent diarrhea, and tropical enteropathy all may contribute to reduced utilization of plant provitamin A. The accepted 6:1 equivalency of beta-carotene to preformed vitamin A must be challenged and reexamined in the context of dietary plants. The consequences of operating on a miscalculation could be serious indeed for public health programs designed to alleviate and eradicate hypovitaminosis A. PMID- 8414224 TI - Lipofuscin, the age pigment. AB - Lipofuscin is the fluorescent mixture of compounds that accumulates increasingly with age in human tissues, including the pigment epithelium of the retina. A substantial component of lipofuscin has been identified in the retina as N retinylidene-N-retinylethanolamine. This compound presumably is derived from the reaction of retinaldehyde with the membrane lipid phosphatidylethanolamine. PMID- 8414225 TI - Food that "disappears" into the food supply: is it all consumed? AB - Trends in food and nutrient intake are estimated by per capita food supply data and surveys of individual consumption. A recent study suggests that these two types of estimates do not reflect the same trends in macronutrient intake. These data should be used with caution when developing associations between diet and risk of disease. PMID- 8414226 TI - Proposed nutrient and energy intakes for the European community: a report of the Scientific Committee for Food of the European community. PMID- 8414227 TI - Calcium and colon cancer. PMID- 8414228 TI - Paroxetine: a new selective serotonin uptake inhibitor for depression. PMID- 8414229 TI - Zolpidem: a nonbenzodiazepine hypnotic. PMID- 8414230 TI - Atovaquone: a new oral treatment for pneumocystis. PMID- 8414231 TI - A comparison of self-reported self-care practices of pregnant adolescents. AB - Recent research has suggested that effective prenatal care is significantly related to positive outcomes for the teen mother and her infant. One aspect of prenatal care is the efforts of the pregnant teen to care for herself, often called self-care practices. The purpose of this article is to compare the self reported self-care activities of pregnant teenagers who sought prenatal care during the first trimester of their pregnancies with those of pregnant teens who delayed prenatal care until the third trimester. The subjects were adolescent girls who were enrolled in prenatal care programs or who had recently delivered in Florida. Interview analysis suggested that both groups had an adequate knowledge level regarding diet, exercise, and other topics related to self-care during pregnancy. Although pregnant teens practice similar self-care behaviors regardless of when they enter prenatal care, there are many problems that may lead to poor pregnancy outcomes and can remain undetected when prenatal care is delayed. PMID- 8414232 TI - Use of terbutaline sulfate in a subcutaneous infusion. Pump to treat preterm labor. AB - The subcutaneous infusion of terbutaline sulfate (Brethine) by an external, portable pump offers an innovative new approach for treating preterm labor. Care of women receiving this therapy modality requires a basic understanding of the pump and its function. Researchers indicate pump tocolysis has numerous advantages over oral tocolysis: daily medication doses are lower, causing fewer side effects; onset of the medication is faster, resulting in the ability to control contractions before cervical changes occur; there is improved patient compliance; and there are higher pregnancy prolongation rates. Successful use of the pump requires in-depth education of the patient and her family. Home support and follow-up are also critical to the success of outpatient management of patients being treated with pump tocolysis. PMID- 8414233 TI - Evaluating suicide risk in the medically ill patient. AB - Common medical conditions, treatments, medications, and depression can increase the risk of suicide, which is the eighth leading cause of death in the United States. Health care providers are in key positions to prevent suicide because most people seek medical treatment for physical symptoms and they give clues before they commit suicide. Unfortunately, suicide risk typically remains undetected because suicide clues, including depression, are overlooked. This article reviews assessment of suicide risk, incidence of completed suicide, and interventions in primary care settings. Suicide lethality, therapeutic strategies, and referrals are considered. Depression is discussed because it is the most common precipitant of suicide in primary care. PMID- 8414234 TI - Certification test development at the American Nurses Credentialing Center. PMID- 8414235 TI - A contemporary view of alternative healing modalities. AB - Alternative forms of healing have increasingly gained popularity in the United States as evidenced by books, healing groups, and a burgeoning variety of healing modalities. Knowledge about these modalities can facilitate health care providers' effectiveness in managing care by understanding the healing practices selected by patients, and incorporating alternative approaches into holistic case management. A matrix is presented that classifies these modalities according to preparation of the healer and methods of healing. A brief description is provided of various types of alternative healing with some of their potential risks and benefits to health. Suggestions are made regarding selection of a healer and sources of information. PMID- 8414236 TI - Learning by degrees. PMID- 8414237 TI - Never alone. PMID- 8414238 TI - Drug alert: packaging leads to fatal errors. PMID- 8414240 TI - A kiss for Mr. Bob. PMID- 8414239 TI - H.I.V.-positive patient. Planning revenge. PMID- 8414241 TI - Documenting your patient's valuables. PMID- 8414242 TI - Your role in intrapleural drug administration. PMID- 8414243 TI - Helping your patient live with osteoporosis. PMID- 8414244 TI - Caring for the depressed patient. PMID- 8414245 TI - Nasal packing: stopping excessive blood loss. PMID- 8414246 TI - Patient assessments: what J.C.A.H.O. requires you to do. PMID- 8414247 TI - Managing pressure ulcers. PMID- 8414248 TI - Preventing falls in the hospital. PMID- 8414249 TI - Traumatic amputation. PMID- 8414250 TI - Three cancer complications that can't wait. PMID- 8414251 TI - Latex allergies and condom use. PMID- 8414252 TI - More on pulmonary edema. PMID- 8414253 TI - Close-up on posterior shoulder dislocation. PMID- 8414254 TI - Caring for culturally diverse patients. PMID- 8414255 TI - The price of heartstrings. PMID- 8414256 TI - Helping patients manage intensified insulin regimens. PMID- 8414257 TI - Skipper's run. PMID- 8414258 TI - Surviving a malpractice lawsuit. One nurse's story. PMID- 8414259 TI - The trouble with sister. PMID- 8414260 TI - IV technology helps you stay out of harm's way. PMID- 8414261 TI - Protecting yourself against T.B. PMID- 8414262 TI - Drug update: T.B., H.I.V., and hepatitis B. PMID- 8414263 TI - Evaluating T.S.H. PMID- 8414264 TI - Myths & facts ... about organ and tissue donation. PMID- 8414265 TI - 8 ways to make yourself more promotable. PMID- 8414266 TI - Controlling a child's pain. PMID- 8414267 TI - Making the most of your resume. PMID- 8414268 TI - The write stuff. PMID- 8414269 TI - Undernutrition in Nigeria: dimension, causes and remedies for alleviation in a changing socio-economic environment. AB - Undernutrition in Nigeria is a long standing problem which has persisted since the 1960s and whose magnitude is on the increase. This is because food consumption, both in quality and quantity, has decreased appreciably, especially with the commencement of the structural adjustment programme (SAP) in 1986. Available studies from limited data have indicated that the introduction of economic reforms more than anything else has contributed to reduced food intake and the near collapse of nutrition oriented health delivery services. Since the economic reforms may continue into the next decade and beyond, sustainable remedies for alleviation of the problem are urgently needed. Suggested remedial programmes include increased support for the rural farmers, strengthening of the rural credit schemes that are specifically targeted at the poor, distribution of vitamin A and iron supplements in rural health centres, encouraging production of low cost weaning diets and integrating nutrition education in primary health care schemes and in educational curricula. PMID- 8414270 TI - The diet of girls and young women at the beginning of the century. AB - Recent research has shown that retarded growth during fetal life and infancy is linked to the development of cardiovascular disease (coronary heart disease and stroke) in adult life. Maternal nutrition has an important effect on early growth and the diets of young women may therefore influence cardiovascular disease in the next generation. Samples of women aged 80 years and over were interviewed in six areas of England and Wales with different cardiovascular death rates. The women, 281 in total, were asked about their diets when they were aged 10 to 15 years. Those who grew up in areas which now have low cardiovascular mortality tended to eat four meals a day rather than three, to live in households which had gardens, kept hens or livestock, and to go into domestic service, where diets were generally good. Those who grew up in areas which now have high cardiovascular mortality tended to eat less red meat, to live in houses without gardens, to enter industrial occupations and have higher fertility rates. PMID- 8414271 TI - Obesity among secondary school students in Bahrain. AB - The aim of this study is to estimate the prevalence of obesity and factors associated with it in Bahraini secondary school students. A cross-sectional study involving a sample of 825 students (417 boys and 408 girls) aged 15 to 21 years was obtained from secondary schools. Obesity was determined using body mass index (BMI = Wt/Ht2). The findings revealed that 15.6% of boys and 17.4% of girls were either overweight or obese (BMI > or = 25). Family size, parents education, and family history of obesity were significantly associated with obesity among boys, while family history was the only socio-economic factors statistically associated with obesity among girls. Meal patterns such as eating between meals, number of meals per day, and method of eating were not associated with obesity in students. Boys who ate alone were 3 times more likely to be obese than those who ate with family members (odd ratio = 3.4). Measures to prevent and control obesity among children are suggested. PMID- 8414272 TI - Breast versus bottle feeding. AB - Significant or highly significant differences were found between groups of breast and bottle-fed infants with respect to the incidence of jaundice, time to passage of first yellow stool and early weight changes. Experimental parameters were carefully defined, and a simple stool colour comparator has been designed and developed. Breast feeding should be encouraged for its incidence is low in many European countries and it is the responsibility of the attending midwife to ensure that correct suckling techniques are achieved by mothers and infants at the earliest possible time after delivery. PMID- 8414273 TI - Antioxidant nutrients and disease. PMID- 8414274 TI - The effects of food shortage on human reproduction. PMID- 8414275 TI - A new approach to Russia's health problems. PMID- 8414276 TI - Chronic fatigue syndrome (ME) PMID- 8414277 TI - Low follicular oestrogen levels in New Zealand women consuming high fibre diets: a risk factor for osteopenia? AB - AIMS: To study effects of anthropometry, diet and exercise on plasma 17 beta oestradiol concentrations in healthy premenopausal women. METHODS: Fifty-five healthy female volunteers aged 19-29 (Mean (SD) = 22.2 (2.6) years) were studied. Anthropometric measurements were made, food intakes were estimated using a food frequency questionnaire, physical activity was assessed, menstrual histories were taken and blood 17 beta-oestradiol values were measured in the follicular and luteal phases of the menstrual cycle. RESULTS: Adiposity and dietary fat intake did not influence circulating plasma 17 beta-oestradiol but dietary fibre intake was negatively correlated with both follicular (r = -0.43, p < 0.001) and luteal (r = -0.28, p < 0.05) 17 beta-oestradiol concentrations. Follicular oestradiol values were abnormally low in 53% of the population. Subjects with high fibre intakes (> or = 25 g fibre daily) had significantly lower concentrations of 17 beta-oestradiol in the follicular phase of the menstrual cycle (p < 0.01). Physical activity was similar in groups consuming high and low fibre diets. Subjects with oligomenorrhoea or amenorrhoea (n = 7) had higher fibre intakes/energy (p < 0.05) than subjects with eumenorrhoea (n = 48). CONCLUSION: Our study suggests that healthy New Zealand women of premenopausal age have lower 17 beta-oestradiol levels on high fibre diets than on low fibre diets. Because hypoestrogenism is a known risk factor for osteoporosis we propose that high fibre intakes may influence bone mass adversely and be a risk factor for osteoporosis. The effects of high fibre intakes on bone mass warrant further investigation. PMID- 8414278 TI - Lead exposure during recreational use of small bore rifle ranges. AB - AIMS: Following detection of symptomatic lead toxicity in two users of an indoor small bore rifle range, we studied users of several similar facilities to determine if significant recreational lead exposure occurred. METHOD: Red cell lead levels were measured at the end of a six month (winter) indoor shooting season and prior to commencement of shooting in the following year. Lead levels in air and dust sampled at one range were also measured. RESULTS: REd cell lead levels were elevated at the end of season (mean 2.64 mumol/L) and lower (mean 1.60 mumol/L) in the preseason samples. The average red cell lead level of the male shooters was 2.4 times normal and is comparable to the levels found in many occupationally exposed groups. Maximum air lead levels were 210 micrograms/m3, more than 2 times the Department of Labour OSH workplace exposure standard TWA of 100 micrograms/m3. Analysis of dust samples showed that dust at this range contained 24% to 36% lead. CONCLUSION: Although the mean time spent shooting was only 70 minutes per week the blood lead levels are similar to those previously reported for full time instructors at pistol ranges. This data confirms that lead exposure in recreational users of indoor small bore rifle ranges is a significant problem. PMID- 8414279 TI - Strengthening the chain of survival. PMID- 8414280 TI - Patterns of smoking during pregnancy in Canterbury. AB - AIMS: To examine the prevalence and patterns of smoking in pregnancy with the object of improving smokefree programmes in the region. METHODS: A postal questionnaire on smoking in pregnancy was sent to all 1916 mothers giving singleton births in the Canterbury region over a five month period. There was a 71.7% response rate, however, smokers were significantly under represented. Data from nonresponders was obtained from obstetric records. RESULTS: Of the total sample, 30% smoked during their last pregnancy. There were significant differences between responders and nonresponders. The responders contained only 60% of all smokers. Nonresponders had twice the incidence of smoking, were more likely to identify as Maori, were younger and had lower birth weights. Nonresponders contained 40% of all smokers. Of the responders, 333 mothers smoked during at least some part of pregnancy: 113 (34%) quit, 168 (50%) cut down, and 48 (15%) made no change. Most (90%) of those who did quit did so during the first trimester. Lighter smokers (less than 10 per day) were more likely to quit or cut down. But smoking rates subsequently increased after the birth. CONCLUSIONS: In Canterbury, 30% of pregnant women smoke. Although 64% indicated a wish to quit and 30% to cut down, this contrasted with what they actually achieved: 34% quit and 50% cut down. Pregnancy influences smoking patterns and is an opportune time for smokefree promotion. PMID- 8414281 TI - Criteria for gestational diabetes: a cautionary tale. AB - The diagnosis of gestational diabetes was developed to predict neonatal outcome (particularly perinatal mortality, macrosomia and hypoglycaemia) and future maternal diabetes. A variety of criteria for this diagnosis have evolved over time, assessed predominantly among European women. We describe a Pacific Islands woman with multiple risk factors for future diabetes yet a borderline 100 g glucose tolerance test result, who delivered a stillborn macrosomic baby weighing 6.7 kg at 38 weeks' gestation. Six weeks postpartum, diabetes was diagnosed by 75 g oral glucose tolerance test. This case highlights the need for caution when interpreting the glucose tolerance test in pregnancy and suggests that closer fetal monitoring and involvement of the diabetes team may be necessary among women with a borderline glucose tolerance test in the presence of additional risk factors for future diabetes (eg obesity, ethnic group). PMID- 8414282 TI - Trainee interns: education and service roles. AB - AIMS: To examine the education and service roles of trainee interns in New Zealand. METHODS: A questionnaire was completed by 123 trainee interns from the four clinical teaching schools. Areas examined were the amount of teaching received and the groups providing teaching, service work load, debt levels and the value of electives. RESULTS: In hospitals trainee interns are taught for 30% of their week. The rest is spent performing service work, 58% of which is unsupervised and 12% supervised. Registrars and house surgeons provide the bulk of total teaching, 33% and 32% respectively. On average, trainee interns in general practice were working 40 hours/week and while on the wards 50 hours/week. On relevant wards trainee interns performed 38% of the ward work. The position of acting house surgeon had been filled by 88% of trainee interns. A debt greater than $10,000 was carried by 40%. Without the trainee intern salary 89% would not have been able to afford their electives, 25% performed electives in New Zealand. Electives were viewed as very beneficial in a number of areas. CONCLUSIONS: Trainee interns are working many hours, providing valuable service work in New Zealand hospitals. The majority of teaching is practical, on the wards, from registrars and house surgeons. Debt affects a substantial number. Electives are a valuable part of the trainee intern course. PMID- 8414283 TI - Biphasic positive airway pressure for airway obstruction secondary to adenotonsillar hypertrophy. PMID- 8414284 TI - Hepatitis C: the tracing dilemma. PMID- 8414285 TI - Spontaneous remission of cancer. PMID- 8414286 TI - Cot death rates amongst Maori. PMID- 8414287 TI - Pregnancy associated with a combined oral contraceptive and itraconazole. PMID- 8414288 TI - Presentation of data. PMID- 8414289 TI - Alcohol and pregnancy. PMID- 8414290 TI - Maternal mortality. PMID- 8414291 TI - Simultaneous small and large bowel ulceration associated with short term NSAID use. PMID- 8414292 TI - Why nurses lose their licenses--Part I. PMID- 8414293 TI - Take the high road, Mr. President. PMID- 8414294 TI - Managing innovative technology. PMID- 8414295 TI - When must others choose? PMID- 8414296 TI - Care manager/nurse manager: a blending of roles. AB - Excellent clinicians often are "promoted" to management positions. Lack of involvement in patient care can lead to diminished nurse manager satisfaction, so job enrichment and enhancement must be as important as efficiency and economy as healthcare undergoes reform. A study of 314 patients from 20 DRG categories revealed a statistically significant shortened length of stay and a savings of at least $552 per care-managed patient. Satisfaction was widespread among nurses, patients and physicians and a collaborative spirit developed between nurses and physicians. The blended care manager/nurse manager role alleviates much frustration and conflict and enriches the performance of the manager. PMID- 8414297 TI - Starting a hospital-based home health agency: Part III--Marketing. AB - Successfully marketing home healthcare involves not only community awareness, but the system's support as well--nurses, physicians, administration, social services. Working together with common goals and commitments is essential to the program's success. Addressing questions and concerns ensures a successful business start-up and ongoing implementation. A service benefit profile, target markets, and a feasibility analysis are provided in this final section of a three part series on establishing a home health agency. PMID- 8414298 TI - Catch 22: manipulative family members. AB - When a Medicare patient from another state, who is not deemed mentally incompetent, vehemently demands treatment but then defers all procedures and treatment to her questionably competent son who, in turn, curtails any progress toward diagnosis, what are the hospital's and attending physicians' rights to continue basic care to the tune of $90,000 and beyond? Such multifaceted and complex dilemmas are illustrated by this case study which outlines the challenges and restraints encountered in dealing with an unusual situation. Also, it offers guidelines as a framework for action when a manipulative family member or patient is encountered. PMID- 8414299 TI - Retention can be improved! AB - A unique program has been developed to encourage retention of nurses who prefer to remain in direct patient care roles. Both recognition and monetary reward are available to nurses who join the "Professional Excellence in Nursing" (PEN) program. The underlying philosophy is that stable employment, clearly defined and attainable goals and recognition for a higher level of performance are keys to job satisfaction--and thus to retention. Evaluative data on this young program are not available but informal surveys indicate a positive outcome. PMID- 8414300 TI - Nurse manager performance appraisal: a collaborative approach. AB - The job description and performance appraisal criteria for the Clinical Nurse Manager (CNM) were revised to provide clarity of role responsibilities and promote collegiality. The new formats reflected much better the complexities and differences existing in the 15 distinct inpatient and ambulatory practice settings. Six categories were defined which summarized the CNM role and a qualitative tool was developed for performance evaluation. PMID- 8414301 TI - Applying a political model to program development. AB - Politics encompasses personal behaviors that dominate much of day-to-day decision making in organizations. Conflict arises when various groups with different goals, interests, values and perceptions attempt to influence policy or decision making to meet their own interests, needs, goals or values. In this paper, Baldridge's political model is used to analyze the conflict experienced in development of a hospital-based discharge planning program. PMID- 8414302 TI - The nursing process: a tool for healing the environment. AB - Assessment, planning, implementation and evaluation can be useful in addressing environmental issues. "Eco-teams" can develop comprehensive waste management programs through implementing a "reduce, reuse, recycle, buy recycled" approach. Serious commitment to such behavior can result in significant savings while reducing dramatically the resources, energy and pollution associated with manufacture and disposal of many items commonly used in healthcare settings. PMID- 8414303 TI - Providing acute geriatric care. PMID- 8414304 TI - Improving patient education documentation. PMID- 8414305 TI - Twelve nurses tell their ethical stories. PMID- 8414306 TI - Managing change: introducing ancillary caregivers to the ICU. AB - How to introduce ancillary care personnel in a critical care setting is chronicled in this article. The goal was to try assigning three patients to one RN working with a critical care nurse technician (CCNT) in an effort to deal with a 30-35 percent vacancy rate in CCU/MICU staffing, which had resulted in closing of beds. The approach to change described by Moore and Gergen enabled this management team to move toward their goal by including both nursing staff and CCNTs in the process. PMID- 8414307 TI - Autonomy through androgyny. AB - The perception of contemporary nursing as being subordinate and subservient to medicine can be compared to the role of chloroform, which in its day served the purpose intended. However, like nursing, today's anesthetic agents require greater expertise to maximize their effectiveness. As nursing enters a new era in its struggle for autonomy, androgynous approaches promise increased independent thinking, action and self-esteem. Nurses possessing these qualities can address effectively, many of the challenges today's nursing practice poses. PMID- 8414308 TI - Stress and the single sybarite. PMID- 8414309 TI - Nursing "cannibalistic" toward its elders, too ... PMID- 8414310 TI - The impact of I. C. Rubin, Md, on obstetrics and gynecology. PMID- 8414311 TI - Antenatal diagnosis of choroid plexus cyst: suggested management. PMID- 8414312 TI - Sexual assault. AB - Sexual assault continues to represent the most rapidly growing violent crime in America. Vital legal reforms are underway, but statistics prove a persistent rise in rape incidence with poor conviction rates. This knowledge, along with the vast multitude of emotional sequelae of rape and self-perceived inferior legal status of women, results in a high percentage of unreported cases. It is imperative that health care providers understand the horrific nature of sexual assault in order to provide appropriate care. All medical care personnel involved in the care of potential rape victims should be briefed in historic and modern legalities of sexual assault. Specific training in emergent and chronic care, both physical and mental, in conjunction with an understanding of rape legislation is vital if health care professionals are to appropriately care for victims of rape. PMID- 8414313 TI - How to increase the cost-effectiveness of IVD programs. PMID- 8414314 TI - The law and digitized images. PMID- 8414315 TI - Primary and preventive care services provided by obstetrician-gynecologists. AB - OBJECTIVE: To determine the level and types of primary and preventive care services delivered by obstetrician-gynecologists. METHODS: A self-administered questionnaire was mailed to a random sample of 1250 obstetrician-gynecologists practicing in the United States. The response rate was 71%. RESULTS: Fifty-three percent of the respondents indicated that they provide primary and preventive care during more than half of their practice time. Although obstetrician gynecologists provide a wide range of preventive services, the proportion of doctors providing any specific service varies. Whereas virtually all (92% or more) obstetrician-gynecologists provide or order blood pressure screening, breast examinations, mammography, and Papanicolaou tests, only six of ten report regular cholesterol screening for most of their patients. A higher percentage of female obstetrician-gynecologists, who are on average younger than their male counterparts, report that they provide primary preventive services to most of their patients. CONCLUSION: The majority of obstetrician-gynecologists provide a wide range of primary and preventive care services to their patients, although there is variability in the proportion of doctors providing any specific service to most (60% or more) of their patients. PMID- 8414316 TI - The accuracy of endometrial Pipelle sampling with and without sonographic measurement of endometrial thickness. AB - OBJECTIVE: To evaluate the accuracy of Pipelle endometrial sampling with and without sonographic measurement of endometrial thickness. METHODS: We studied prospectively 176 consecutive patients (23% after and 77% before menopause) scheduled for D&C. Sonographic measurement of the endometrium and endometrial biopsy with the Pipelle were performed before the curettage. RESULTS: In 159 cases (90%), the endometrial histologic results of curettage agreed with those of the Pipelle biopsy. All three cases of endometrial cancer were identified by Pipelle aspiration. In seven cases (4%), the Pipelle aspiration failed to detect hyperplasia. Sonographic endometrial thickness of more than 5 mm slightly increased the sensitivity and slightly decreased the specificity of Pipelle aspiration from 82 to 92% and from 99 to 96%, respectively. In postmenopausal patients admitted for bleeding, the sensitivity and specificity reached 100%. CONCLUSIONS: Our data suggest that normal Pipelle aspirates in premenopausal patients with abnormal uterine bleeding are highly accurate. In postmenopausal patients with sonographic endometrial thickness of 5 mm or less, the accuracy reached 100%. PMID- 8414317 TI - The use of large-loop excision of the transformation zone in an inner-city population. AB - OBJECTIVE: To determine whether large-loop excision of the transformation zone (LLETZ) can be used in our population to treat patients in a routine colposcopy clinic without diminishing the capability to see the large number of patients who require care, and to compare these results to our previous experience with laser surgery. METHODS: Patients found to have squamous intraepithelial lesions on colposcopically directed biopsies were offered treatment with LLETZ during counseling regarding their biopsy findings. Procedures included "ablation equivalents" and "cone biopsy equivalents" using local anesthesia. Follow-up examinations were used to determine cure and included cytology, colposcopy, and directed biopsies when indicated. The cure rate was compared to our previous experience with laser surgery. RESULTS: Two hundred thirty-six patients were treated in the colposcopy clinic without diminishing the capability to see all patients requiring care. Complications were few. The mean (+/- standard deviation) follow-up period was 50.7 +/- 25.3 weeks. The overall cure rate of 91.3% (95% confidence interval [CI] 87.1-95.5) was not influenced by the severity of the disease, but positive endocervical margins significantly lowered the cure rate to 69.2% compared with those who had negative margins. Before this series, only 73.1% of our patients scheduled for laser surgery returned for treatment. Assuming a 90% cure rate among those who returned, this means that the actual cure rate was only 65.8%. The likelihood of cure was 1.37 times greater (95% CI 1.27-1.52; P < .0001) using LLETZ in the clinic at the time the patient was counseled regarding her biopsy findings than using laser at a later date. In 33 patients, the LLETZ specimen showed no evidence of disease. The relative risk of negative histology was 3.31 (95% CI 1.78-6.13; P < .001) when LLETZ was done for a discrepancy between cytology and histology as opposed to any other indication. Cancer was found on the LLETZ specimen in four patients (two microinvasive, two frankly invasive), but was not suspected preoperatively in any of the patients. CONCLUSION: In our inner-city clinic, treatment with LLETZ at the time the patient was counseled regarding her biopsy findings improved the actual cure rate. The LLETZ procedure can be done safely in a clinic setting without diminishing the capability to care for a large number of patients. PMID- 8414318 TI - Transcervical hysteroscopic resection of submucous fibroids for abnormal uterine bleeding: results regarding the degree of intramural extension. AB - OBJECTIVE: To examine the results of transcervical resection of submucous fibroids in relation to the degree of intramural extension. METHODS: A prospective 3-year observational study was performed of transcervical resection of submucous fibroids for abnormal uterine bleeding. The mean follow-up was 20 months (range 10-34). Fifty-one patients with a mean age of 38 years (range 23 55) were treated with transcervical resection after classification according to the degree of intramural extension of the submucous fibroids. The intention was to perform complete resection, established at control hysteroscopy. A repeat procedure was performed in cases of incomplete resection unless the patient denied further hysteroscopic treatment. Outcome measures were control of bleeding, subsequent surgery, number of procedures, number of complete resections, and number of recurrences. RESULTS: Bleeding was controlled in 48 (94.1%) of all patients after final resection. Hysterectomy was performed in three patients (5.9%) because of persistent complaints: in two cases after incomplete resection and in one case after complete resection. Three patients were lost to follow-up. Of the remaining 45 patients (42 with complete and three with incomplete final resection), three (6.7%) had a recurrence (one after complete and two after incomplete final resection). With more extensive intramural involvement, the chance to achieve complete resection decreased and the mean number of procedures to achieve complete resection increased. CONCLUSIONS: Complete resection improves the long-term results of transcervical resection of submucous fibroids for control of abnormal uterine bleeding. Transcervical resection of submucous fibroids with more than 50% intramural extension should be performed only in selected cases, as complete resection usually necessitates repeat procedures. PMID- 8414319 TI - Laparoscopic para-aortic lymphadenectomy in gynecologic malignancies. AB - OBJECTIVE: To determine the feasibility, safety, limiting factors, and advantages of laparoscopic para-aortic lymphadenectomy in a series of patients with gynecologic malignancies. METHODS: During a 2-year period, 61 women underwent laparoscopic para-aortic lymph node dissection as part of their management for invasive gynecologic malignancies. A transperitoneal incision directly over the aorta was used. Initially, only the right-side infra-inferior mesenteric artery nodes were removed. The technique of removal of left-side low para-aortic nodes was then developed, followed by the technique for removal of right- and left-side nodes above the transverse duodenum. A total of 52 right para-aortic lymphadenectomies were performed, 12 of which were combined with left-side lymphadenectomies. A total of 17 left-side lymphadenectomies were performed, 12 of which were bilateral. Four patients had nodes removed above the inferior mesenteric artery. RESULTS: The procedure could not be performed in four instances because of obesity or adhesions. Twenty-four patients had their laparoscopic surgery combined with another procedure, which increased their hospital stays: radical hysterectomy (five), laparoscopy-assisted vaginal hysterectomy (17), transperineal interstitial irradiation (one), and anterior posterior colporrhaphy (one). The remaining 33 patients had laparoscopic surgical staging only. One patient required laparotomy to control bleeding from the vena cava; however, the others had no short- or long-term complications, and the average hospital stay was 1.3 days. CONCLUSION: Laparoscopic para-aortic lymphadenectomy is a safe, effective procedure that allows a shorter hospitalization than traditional laparotomy. PMID- 8414320 TI - Second-look laparotomy in carcinoma of the fallopian tube. AB - OBJECTIVE: To evaluate the role of second-look laparotomy following platinum based chemotherapy in patients with fallopian tube cancer. METHODS: We conducted a retrospective chart review of 35 patients with tubal cancer who underwent a second-look laparotomy following cytoreductive surgery and platinum-based combination chemotherapy. RESULTS: The distribution by stage was as follows: I, three (9%); II, five (14%); III, 24 (69%); and IV, two (6%); one patient was inadequately staged. The tumor grade was recorded in 31 patients, and 80% of these were grade 2 or 3. Twenty-one of the 35 patients (60%) were found to be free of disease at second-look operation. Neither stage nor grade were predictive of findings at the second-look procedure, although none of the five patients with stage I disease and/or grade 1 tumor had disease found at second-look surgery. The absence of gross residual disease following primary surgery was the strongest predictor of disease-free status at second-look laparotomy (P < .01). With a mean follow-up of 50 months among the survivors, only four (19%) of the patients with no evidence of tumor at reexploration have had a recurrence. CONCLUSION: Second look laparotomy provides useful prognostic information in patients with tubal cancer; approximately 80% of patients who have a negative second-look following platinum-based chemotherapy will remain disease-free. PMID- 8414321 TI - Borderline and invasive epithelial ovarian tumors in young women. AB - OBJECTIVE: To review the occurrence, morbidity, and mortality of borderline and invasive epithelial ovarian tumors in young women. METHODS: We conducted a 15 year retrospective review of the case records of the Women's Cancer Center, University of Minnesota, and the JL McKelvey Tumor Registry. RESULTS: We identified 67 patients under age 40 with borderline or invasive epithelial ovarian tumors. Fifty patients (75%) had borderline tumors and 17 (25%) had invasive tumors. The mean age at presentation was 31 years (range 14-39) for the borderline group and 34 years (range 23-39) for the invasive group. Pelvic pain and a palpable mass, present for less than 6 months, were the predominant presenting symptom and sign. There was no difference in the age at menarche between the patients with borderline (mean 12 years) and invasive tumors (mean 13 years). Fifty-seven patients were optimally cytoreduced to less than 2.0 cm after primary surgery. Thirty-five patients underwent second-look laparotomy, 15 of which were positive for tumor. A minority of patients in both groups had stage I tumors (17 in the borderline and one in the invasive group). Among patients with borderline tumors, there was no difference between "very young" and "young" patients in the stage at presentation or outcome. Similar proportions of patients presented with early- and late-stage disease. Three very young women (14%) and five young women (17%) have died. Among patients with invasive tumors, no difference existed between young and very young patients for stage at presentation, whereas grade and outcome differed significantly between the age groups (P < .05). Very young patients were more likely to present with grade 1 lesions, whereas patients aged 30-40 years were more likely to have grade 2 or 3 tumors (P < .05). Three (100%) of the very young patients have died, whereas seven (50%) of the young patients aged 30-40 years have died. The median survival of patients with borderline tumors was 36 months (range 2.0-150.5), significantly different from those with invasive tumors, whose median survival was 21 months (range 2.9-89.7) (P < .001). CONCLUSION: Borderline and invasive epithelial ovarian tumors are encountered in young women. Despite the implication of the term "borderline," such tumors are associated with considerable morbidity and mortality. PMID- 8414322 TI - Hysterectomy and race. AB - OBJECTIVE: To investigate black-white differences in factors related to hysterectomy. METHODS: Discharge summary data were analyzed for 53,159 hysterectomies that occurred in Maryland from 1986-1991. RESULTS: The average annual age-adjusted hysterectomy rate was higher for black women (49.5 per 10,000) than for white women (41.2 per 10,000). For 65.4% of the hysterectomies in black women, the principal diagnosis was uterine fibroids, compared to 28.5% for white women. Logistic regression was used to measure the effect of race on complications, length of stay, and mortality after adjustment for a variety of factors including age, comorbidities, diagnosis, route (abdominal, vaginal, or subtotal), hospital characteristics, and source of payment. In comparison to white women, black women having hysterectomy were found to have an increased risk of one or more complications of surgical or medical care (odds ratio 1.4, 95% confidence interval [CI] 1.3-1.5), a length of stay of more than 10 days (odds ratio 2.7, 95% CI 2.5-3.1), and in-hospital mortality (odds ratio 3.1, 95% CI 2.0 4.8). CONCLUSIONS: In a study of more than 53,000 hysterectomies, black women were more than twice as likely to have a diagnosis of uterine fibroids as white women, were more likely to have complications, had a longer hospitalization, and had more than three times the in-hospital mortality rate. PMID- 8414323 TI - Pelvic inflammatory disease in human immunodeficiency virus-infected women. AB - OBJECTIVE: To determine whether the clinical course of pelvic inflammatory disease differs between women with human immunodeficiency virus (HIV) infection and seronegative controls. METHODS: All admissions for acute pelvic inflammatory disease from January 1, 1986 to December 31, 1992 at San Francisco General Hospital were reviewed, identifying 23 HIV-seropositive women. Their clinical course was compared with a control group of 108 seronegative women admitted for acute pelvic inflammatory disease. RESULTS: Human immunodeficiency virus seropositive women with acute pelvic inflammatory disease had significantly lower abdominal tenderness scores (P < .05), lower admission and discharge white blood cell counts (WBC) (P < .01, P < .05), and fewer gonococcal infections (odds ratio 0.3, 95% confidence interval [CI] 0.1-0.9; P < .05) than the seronegative controls. There were no significant differences in duration of treatment, length of hospitalization, or incidence of tubo-ovarian abscess. Significantly more HIV positive women with acute pelvic inflammatory disease required surgical intervention than seronegative women (odds ratio 5.5, 95% CI 1.0-29.3; P < .05). CONCLUSION: Human immunodeficiency virus-seropositive women with acute pelvic inflammatory disease may have an altered immune response, resulting in inadequate response to antimicrobial agents and the need for more surgical intervention. Future studies must include larger numbers of HIV-infected women, and the results must be stratified for CD4 counts, clinical HIV stage, and other measures of immunocompromise. PMID- 8414324 TI - Treatment of menstruation-associated migraine headache with subcutaneous sumatriptan. AB - OBJECTIVE: To evaluate the efficacy and safety of sumatriptan, a 5-HT1 receptor agonist, in patients with menstruation-associated migraine. METHODS: Two double blind, placebo-controlled, single-attack parallel group studies of subcutaneous sumatriptan were conducted for the acute treatment of migraine. A retrospective analysis of 1104 patients produced 157 women who were treated for a menstruation associated migraine (defined as a migraine beginning between 1 day before and 4 days after the onset of menstrual flow) and 512 women treated for nonmenstrual migraine. We excluded 435 other patients who were either male (123), women with hysterectomies (260), or women with missing data (52). Patients with moderate or severe pain were treated with 6 mg subcutaneous sumatriptan or placebo. One hour after treatment, response rates of headache severity and associated symptoms were measured. Menstruation-associated migraine patients were compared to female patients with nonmenstrual migraine. Migraine recurrence was analyzed retrospectively for 24 hours. RESULTS: At 1 hour, 80% of the sumatriptan-treated menstrual-migraine patients had pain relief (reduction of severe or moderate pain to mild or no pain), compared to 19% of the placebo patients (P < .001). Sumatriptan also treated nausea and photophobia more effectively in menstrual migraine patients than did placebo. Response rates for pain and associated symptoms were similar between patients with menstruation-associated and nonmenstrual migraines. Adverse effects were also similar between the groups. CONCLUSION: Sumatriptan was as effective and well tolerated for menstruation associated migraine as it was for nonmenstrual migraine. PMID- 8414325 TI - Comparison of transdermal and oral sequential gestagen in combination with transdermal estradiol: effects on bleeding patterns and endometrial histology. AB - OBJECTIVE: To assess prospectively the effect on bleeding patterns, transformation of the endometrium, and rate of endometrial hyperplasia of transdermal norethisterone acetate when administered sequentially in combination with transdermal estradiol (E2), and to compare it to a regimen using oral medroxyprogesterone acetate. METHODS: Two hundred eighteen women were randomized to receive either transdermal E2 0.05 mg/day for 14 days followed by transdermal E2 0.05 mg/day plus transdermal norethisterone acetate 0.25 mg/day for 14 days, or transdermal E2 0.05 mg/day for 25 days with the addition of oral medroxyprogesterone acetate 10 mg/day on days 16-25, followed by a 3-day treatment-free period. Treatment was planned for 13 cycles of 28 days. The subjects kept daily bleeding diaries, and endometrial biopsies were taken before and at the end of treatment. RESULTS: The mean duration of bleeding (regular) induced by the gestagen was 7.33 days in the transdermal gestagen-treated group, which was 1.54 days longer than in the oral gestagen-treated group (P = .0001). The mean cycle day of onset was 25.74. Bleeding was spotting or light in the transdermal group in 77% of the days in which bleeding occurred. When comparing the two groups, there were no differences in the overall mean cycle day of onset or in the intensity of gestagen-induced bleeding. Breakthrough (irregular) bleeding episodes occurred in 42% of the transdermal subjects, lasted a mean of 4.18 days, and were spotting or light in 87% of the days when they occurred. There were no differences between the treatment groups. There was only one case (1.3%) of confirmed simple hyperplasia and five cases of failure of gestagenic transformation of the endometrium in 77 women treated for a mean of 367 days with transdermal gestagen. CONCLUSION: A transdermal system delivering a combination of E2 and norethisterone acetate for 14 days in sequence with E2 delivered transdermally for 14 days produced bleeding patterns that are clinically acceptable and comparable to those produced by oral medroxyprogesterone acetate given in sequence with E2 delivered transdermally for 25 days. The use of the combination system was not associated with a significant risk of endometrial hyperplasia. PMID- 8414326 TI - A randomized and longitudinal study of human menopausal gonadotropin with intrauterine insemination in the treatment of infertility. AB - OBJECTIVE: To study in a randomized and longitudinal manner the efficacy of human menopausal gonadotropin (hMG) superovulation combined with intrauterine insemination (IUI) versus IUI alone in the treatment of various causes of infertility in the presence of normal ovulation. METHODS: An initially randomized and subsequently longitudinal study of infertile couples was performed at a university-based clinical research center. One hundred nineteen couples with longstanding infertility (average duration 3.7 years) associated with male factor infertility, unexplained infertility, and/or endometriosis were enrolled. All patients were randomized in the initial cycle to treatment with either hMG/IUI or urine LH-timed IUI alone. They were then followed longitudinally as they alternated subsequent cycles between the two modalities. Outcome indices measured were cycle fecundity, pregnancy outcome, and cumulative pregnancy rates evaluated by life-table analysis. RESULTS: Human menopausal gonadotropin/IUI therapy was consistently more effective than IUI alone in the treatment of endometriosis, male factor infertility, and unexplained infertility, with cycle fecundities ranging from 7.1-19.0% versus 0-6.7%, respectively, during the first seven cycles. CONCLUSION: Human menopausal gonadotropin/IUI is a more effective therapy for enhancing fertility than is IUI alone for the treatment of endometriosis, male factor infertility, and unexplained infertility. PMID- 8414327 TI - Measles in pregnancy: a descriptive study of 58 cases. AB - OBJECTIVE: To describe the effects of measles in pregnancy using a large case series. METHODS: Pregnant women with measles were identified by county health department records, and their hospital and clinic records were reviewed. When available, records for the infants of case patients were also reviewed. RESULTS: Fifty-eight pregnant women with measles were identified. Thirty-five (60%) were hospitalized for measles, 15 (26%) were diagnosed with pneumonia, and two (3%) died of measles complications. Excluding three induced abortions, 18 pregnancies (31%) ended prematurely; five were spontaneous abortions and 13 were preterm deliveries. All but two of the 18 pregnancies that terminated early did so within 14 days of rash onset. Two term infants were born with minor congenital anomalies, but their mothers had measles late in the third trimester. No newborns were diagnosed with congenital measles. CONCLUSIONS: The incidence of death and other complications from measles during pregnancy may be higher than expected for age-comparable, nonpregnant women. Measles in pregnancy may lead to high rates of fetal loss and prematurity, especially in the first 2 weeks after the onset of rash. PMID- 8414328 TI - Early-pregnancy proteinuria in diabetes related to preeclampsia. AB - OBJECTIVE: To test the hypothesis that the risk of preeclampsia in diabetic mothers is increased with incipient diabetic nephropathy as well as with overt nephropathy. METHODS: Pregnancy outcome was studied in 311 women with class B-RF diabetes from two institutions. Using 104 women without chronic hypertension followed at the University of California, San Francisco, we constructed a receiver-operating characteristic curve relating 24-hour urinary total protein before 20 weeks' gestation to the subsequent development of preeclampsia. From the curve, a predictive cutoff level of proteinuria was selected and tested in two validation groups not used to construct the curve: 158 women without chronic hypertension followed at the University of Cincinnati and 49 women with chronic hypertension from both institutions. RESULTS: The receiver-operating characteristic curve showed an increased risk of preeclampsia with early pregnancy proteinuria of 190 mg/day or more. In the Cincinnati validation group, the rate of preeclampsia was 7% in women with early-pregnancy proteinuria of less than 190 mg/day, 31% with proteinuria of 190-499 mg/day, and 38% with proteinuria of 500 mg/day or more. In the chronic-hypertension validation group, the rates were 0, 50, and 58%, respectively. By multiple logistic regression, the increased risk of preeclampsia with proteinuria above 190 mg/day persisted after controlling for the effects of parity, chronic hypertension, retinopathy, and glycemic control. CONCLUSIONS: Diabetic gravidas with early-pregnancy proteinuria of 190-499 mg/day are at increased risk for preeclampsia. The risk is comparable to that in women with overt diabetic nephropathy and is independent of chronic hypertension. We speculate that diabetic women with proteinuria in this range have incipient or subclinical diabetic nephropathy. PMID- 8414329 TI - Maternal serum CA 125 levels in the diagnosis of abruptio placentae. AB - OBJECTIVE: To evaluate measurements of maternal serum CA 125 for the diagnosis of abruptio placentae. METHODS: This investigation was conducted at Swedish Medical Center/Seattle from December 1991 through April 1992. During the study period, we identified 21 consecutive patients with a clinical diagnosis of abruptio placentae. In addition, we enrolled two groups of patients whose pregnancies were not complicated by placental abruption (28 preterm controls and 53 term controls). RESULTS: Mean maternal serum levels of CA 125 were significantly higher among women with abruptio placentae (40.6 +/- 29.6 U/mL) than among preterm controls (26.6 +/- 17.2 U/mL) or term controls (22.0 +/- 11.4 U/mL). Using a positive threshold of 35 U/mL, the sensitivity and specificity of maternal serum CA 125 for abruptio placentae were 42.9% (95% confidence interval [CI] 21.7-64.1) and 92.5% (95% CI 85.4-99.6), respectively. CONCLUSIONS: These findings support an earlier report documenting higher mean maternal serum levels of CA 125 among women with pregnancies complicated by abruptio placentae than in control subjects. However, measurement of maternal serum CA 125 does not appear to be a useful marker for the diagnosis of abruptio placentae. At present, clinical diagnosis of abruptio placentae, with the aid of ultrasonography to rule out other causes of late gestational hemorrhage, should be considered the most sensitive and specific method of detecting this pregnancy complication. PMID- 8414330 TI - Fetal urinary biochemistry predicts postnatal renal function in children with bilateral obstructive uropathies. AB - OBJECTIVE: To investigate the ability of fetal urinalysis to predict in utero the renal function of children with bilateral uropathy who survive to the second year of life. METHODS: This was a prospective cohort study of 100 consecutive patients with prenatal diagnosis of bilateral uropathy who underwent fetal urine sampling. Fetal urinary concentrations of sodium, chloride, calcium, phosphorus, ammonium, urea, creatinine, glucose, proteins, and beta 2 microglobulin were measured. Prenatal findings were matched with renal function of survivors at 1-2 years. The single end point was serum creatinine, which was considered abnormal when greater than 50 mumol/L (0.56 mg/dL) during the second year of life. RESULTS: Elevated serum creatinine was found in 17 of 42 children with isolated uropathy who survived more than 1 year. For prediction of elevated serum creatinine during the second year of life, the fetal urinary concentration of beta 2 microglobulin was both specific (0.83) and sensitive (0.80); sodium, chloride, and urea levels were sensitive (0.70 or greater) but lacked specificity (less than 0.65); and fetal urinary glucose, phosphorus, calcium, ammonium, and total proteins were specific (0.70 or greater) but lacked sensitivity (0.65 or less). CONCLUSIONS: Our results provide a new approach to prenatal management of congenital obstructive uropathies by identifying those fetuses at risk for survival with suboptimal renal function. These fetuses might benefit from intrauterine therapy. In contrast, no attempt at prenatal uro-amniotic shunting should be made when a spontaneously good outcome is predicted by fetal urinalysis. PMID- 8414331 TI - Elevated first-trimester serum relaxin concentrations in pregnant women following ovarian stimulation predict prematurity risk and preterm delivery. AB - OBJECTIVE: To determine whether ovarian stimulation would result in higher circulating relaxin concentrations and whether this hyperrelaxinemia would be associated with prematurity. METHODS: Two groups of women were studied: 1) women achieving pregnancy after ovarian stimulation (n = 114) and 2) women achieving pregnancy without treatment (n = 37). Serum was obtained at 6-12 weeks' gestational age; fetal number was determined by transvaginal ultrasound. Prematurity risk or preterm delivery was determined from the obstetric record. A specific human relaxin enzyme-linked immunosorbent assay was used to measure serum relaxin concentrations. Hyperrelaxinemia was defined as levels greater than 3 standard deviations above the weighted mean of levels in normal unstimulated singleton pregnancies at 6-12 weeks' gestation. RESULTS: An association was found between prematurity risk or premature delivery and peripheral relaxin concentrations during weeks 6-12 of pregnancy in women having ovarian stimulation and in women having multiple gestations. Circulating relaxin concentrations greater than 16 ng/mL in women having ovarian stimulation and levels greater than 7 ng/mL in women who had multiple gestations predicted prematurity risk or premature delivery in 50% of the women. CONCLUSIONS: These data demonstrate that after ovarian stimulation, some women have highly elevated circulating first trimester relaxin concentrations. First-trimester hyperrelaxinemia identifies a group of women at risk for prematurity who can be monitored aggressively. PMID- 8414332 TI - Evaluation of threatened preterm delivery by transvaginal ultrasonographic measurement of cervical length. AB - OBJECTIVE: To establish a nomogram for the length of the uterocervical canal and to determine whether this can be used to predict preterm delivery. METHODS: Cervical length was measured by transvaginal ultrasonography in 32 women (21 primigravid, 11 multigravid) with threatened preterm delivery, and in 177 normal singleton pregnancies between 18-37 weeks' gestation. Regression analysis was used to create the nomogram. Student t test was used to compare the groups. RESULTS: A linear relationship was found between cervical length and gestational age (r = -0.4, P < .001). Comparison of cervical length on admission in the patients with threatened preterm delivery showed that 11 preterm deliveries occurred in women who had a mean cervical length of 23.2 mm (range 17-29), whereas 21 term deliveries occurred in women with a mean cervical length of 31.7 mm (range 21-42). The difference was significant (P < .001). A cervical length of less than 20 mm on admission had a positive predictive value of 100%. These patients had preterm delivery despite tocolytic therapy during hospitalization. CONCLUSION: The risk of preterm delivery is high in women whose cervical length on admission is less than 30 mm, and strict management is required for those with a cervical length of less than 20 mm. PMID- 8414333 TI - Magnetic resonance imaging to avoid laparotomy in pregnancy. AB - OBJECTIVE: To determine whether magnetic resonance imaging (MRI) in pregnancy would help define a benign pelvic mass, thereby avoiding laparotomy. METHODS: During a 2-3-year period, five pregnant women with adnexal masses suspected to be leiomyomas underwent MRI. RESULTS: Four patients had evidence of leiomyoma and one had a benign cystic teratoma. All avoided laparotomy because of the almost certain radiologic findings of a benign process. In two women, there was an important effect on the pregnancy; one had fetal growth retardation and the other an outlet obstruction precluding vaginal delivery. Four of the patients underwent cesarean delivery. CONCLUSIONS: Magnetic resonance imaging can be used in the differential diagnosis of an adnexal mass in pregnancy. This will enable some pregnant patients to avoid laparotomy and its concomitant risks. PMID- 8414334 TI - Comparison of maternal serum lipids before and during parturition. AB - OBJECTIVE: To investigate peripartum progesterone metabolism by measuring its lipid precursors. METHODS: Serum was collected from eight pregnant women not in labor during the third trimester and again during active labor. Samples were analyzed using quantitative spectrophotometric absorbance techniques for apolipoprotein A-1, high-density lipoprotein cholesterol (HDL), and total cholesterol. RESULTS: Remote from labor, the mean gestational age antepartum was 35.6 weeks; during active labor it was 39.2 weeks. The amount of apolipoprotein A 1 present was 182.0 mg/dL before labor and 156.8 mg/dL during active labor; HDL cholesterol was 59.3 mg/dL before labor and 37.5 mg/dL during active labor, and total cholesterol was 226.6 mg/dL before labor and 144.1 mg/dL during active labor. All differences were significant at P < .05. CONCLUSION: Maternal serum lipids associated with progesterone metabolism appear to decrease in the peripartum period. Progesterone metabolism and parturition may be correlated with these lipid changes, thus identifying patients about to enter labor. PMID- 8414335 TI - Treatment of twin-twin transfusion syndrome. AB - OBJECTIVE: To determine whether therapeutic amniocentesis may improve outcomes in patients with twin-twin transfusion syndrome. METHODS: Thirteen patients with possible twin-twin transfusion syndrome were evaluated for treatment. Therapeutic amniocenteses were performed on nine, and four patients were managed conservatively depending on the clinical severity of the twin-twin transfusion syndrome. RESULTS: Therapeutic amniocenteses resolved the syndrome in three of nine cases, with an overall neonatal survival rate of 83.3% (15 of 18) and neonatal morbidity of 53.3% (eight of 15) among the survivors. The survival rate in patients with expectant management was 75% (six of eight), with a neonatal morbidity of 33.3% (two of six). An association between amniotic fluid status and fetal outcomes was observed. Patients with normalization of polyhydramniosoligohydramnios had the best outcomes. CONCLUSION: Early, aggressive amniocentesis may be an effective therapy for twin-twin transfusion syndrome. Therapeutic amniocentesis may have the capability to alter inter-fetal blood flow, possibly as a result of changes in intravascular pressure, which are related to changes in intra-amniotic pressure. PMID- 8414336 TI - Cephalic replacement for shoulder dystocia: present status and future role of the Zavanelli maneuver. AB - OBJECTIVE: To describe the pertinent clinical data, outcomes, and maternal and fetal complications of cephalic replacement for the management of shoulder dystocia. METHODS: A registry for the reporting of cases of cephalic replacement was established 5 years ago. Features of the registry cases were reviewed. RESULTS: Fifty-nine women underwent replacement of the fetal head following unsuccessful attempts at vaginal delivery. All but six infants were successfully replaced and delivered by cesarean without excessive maternal or fetal morbidity. CONCLUSION: Cephalic replacement is a useful technique that need not be used as a last resort, but may be considered if any undue difficulty is encountered. It may have a place as an initial technique for those inexperienced with shoulder dystocia treatment. PMID- 8414337 TI - Effects of carbon dioxide-saturated normal saline and Ringer's lactate on postsurgical adhesion formation in the rabbit. AB - OBJECTIVE: To evaluate the effect of combining carbon dioxide gas (CO2) with normal saline versus CO2 with lactated Ringer's solution on adhesion formation in the rabbit model. METHODS: Sixty New Zealand white rabbits underwent surgery based on a proven experimental adhesion model. Following abdominal closure, the animals were randomly assigned to three groups: Group 1 underwent abdominal CO2 insufflation only; group 2 underwent abdominal irrigation with CO2-saturated normal saline; group 3 underwent abdominal irrigation with CO2-saturated lactated Ringer's solution. Three weeks later, the rabbits were sacrificed and the adhesions were scored in a blinded fashion based on the extent, type, and tenacity, with a maximum possible score of 11. RESULTS: The mean (+/- standard deviation) adhesion scores were 7.75 +/- 2.82 in group 1, 7.85 +/- 2.58 in group 2, and 4.75 +/- 2.95 in group 3. There was no difference in severity of adhesions between groups 1 and 2. However, the mean adhesion score was significantly lower in group 3 (lactated Ringer's with CO2) than in either group 1 (CO2) or group 2 (normal saline with CO2) (P = .004 and P = .002, respectively). CONCLUSION: It appears that when CO2 is the insufflating gas, lactated Ringer's solution has a protective effect against adhesion formation in the rabbit model. PMID- 8414338 TI - Immunization as therapy for recurrent spontaneous abortion: a review and meta analysis. AB - OBJECTIVE: To determine the efficacy of immunotherapy as treatment for recurrent spontaneous abortion (three or more consecutive abortions without an intervening pregnancy of more than 20 weeks' gestation) using meta-analysis. DATA SOURCES: The search began with MEDLINE and was supplemented by reference lists from original research, review articles, and textbooks. We also corresponded with investigators and consulted experts in the field of infertility. METHODS OF STUDY SELECTION: We identified four randomized controlled trials and 19 case-series reports that examined the use of immunotherapy for recurrent spontaneous abortion. DATA EXTRACTION AND SYNTHESIS: A meta-analysis performed on four randomized controlled trials of leukocyte immunotherapy or trophoblast membrane infusion revealed a fixed-effects model odds ratio of 1.3 (95% confidence interval [CI] 0.77-2.3) and a random-effects model odds ratio of 1.3 (95% CI 0.44 3.8). CONCLUSION: The quality of evidence regarding this treatment is class I (best), and the strength of recommendation is D (fair evidence against its use). Unless its efficacy can be established through other randomized controlled trials, this treatment should be abandoned. PMID- 8414339 TI - Whither ultrasonic prediction of fetal macrosomia? AB - During the past 15-20 years, ultrasonic estimation of fetal weight (EFW) has been used increasingly to make management decisions regarding the induction of labor or delivery route. The propriety of this approach depends on proof that its use improves newborn or maternal outcome without disproportionate increases in morbidity and mortality. A barrier to achieving this goal is the inaccuracy associated with ultrasonic EFW. The current ultrasonic EFW procedures are not accurate enough for detecting macrosomia defined by weight criteria. Even if clinicians could determine fetal weight accurately, the frequency of persistent fetal injuries associated with vaginal birth of the macrosomic fetus is so low that induction of labor or cesarean birth is not justified on that basis. Furthermore, the inaccurate ultrasonic determination of fetal weight leads to inappropriate obstetric interventions. Evidence-based decision-making should be an important goal for all obstetric care givers. Delivery decisions based on inaccurate EFW should be avoided. PMID- 8414340 TI - Pelvic floor evaluation with dynamic fluoroscopy. AB - OBJECTIVE: To evaluate dynamic fluoroscopy of the pelvic floor for the study of women with pelvic floor disorders. METHODS: In a prospective, observational study in a tertiary care referral center, 30 women with prolapse beyond the introitus underwent comprehensive fluoroscopic imaging of the pelvic floor. RESULTS: Dynamic fluoroscopy of the pelvic floor was technically possible in all patients. Whereas the physical examinations appeared relatively similar in these patients, the fluoroscopic examination revealed distinct differences. Of the 30 women, 25 had a cystocele, 25 had a rectocele, and 26 had an enterocele. Eleven patients had their surgical plan modified accordingly. Pelvic floor fluoroscopy is not a test for urinary or fecal incontinence. However, incontinence was demonstrated objectively in ten women (eight with fecal incontinence, two with urinary incontinence). CONCLUSION: Dynamic fluoroscopy of the pelvic floor is a useful adjunct in the clinical evaluation of women with prolapse. This imaging can be used to individualize the operative approach to prolapse. It is superior to clinical examination for the detection of enterocele formation. In addition, it provides information regarding the emptying function of the rectocele, which is not obtainable on physical examination. PMID- 8414341 TI - Digital imaging colposcopy: basic concepts and applications. AB - The technique of colposcopy has changed little since the introduction of the green filter to improve viewing of vascular structures. However, the recent dramatic improvements in computer technology now make it practical to combine computerized image processing with colposcopy, which we have termed "digital imaging colposcopy." Image processing techniques permit contrast enhancement of features, such as white epithelium and abnormal vasculature. The digital colposcope allows rapid measurement of the cervical area involved with visible lesions, providing an improved means of following the progression of these lesions over time. PMID- 8414342 TI - Sacrospinous suspension made easy. AB - An orthopedic instrument, the Shutt Suture Punch System, has allowed rapid (less than 1 minute) and safe introduction of the suspending suture through the sacrospinous ligament, especially in the morbidly obese patient. Residents-in training have performed the sacrospinous vaginal suspension on 30 patients without complications. The remaining aspects of the operation are unchanged. PMID- 8414343 TI - Gaining access to the embryonic-fetal circulation via first-trimester endoscopy: a step into the future. AB - OBJECTIVE: To assess the feasibility of gaining access to the embryonic-fetal circulation via first-trimester transcervical embryoscopy. METHODS: A fiberoptic endoscope with a 3.5-mm, wide-angle lens was passed transcervically through the chorion and into the exocoelomic cavity of women undergoing pregnancy termination. A 26-gauge heparinized needle was passed through the sideport of the endoscope and inserted into the blood vessels of the chorionic plate or umbilical cord to obtain blood samples. RESULTS: With a modified endoscope, we have been able to gain access into the embryonic-fetal circulation and obtain a small aliquot of blood in five of eight cases. CONCLUSIONS: Our experience establishes the feasibility of gaining access to the human embryonic-fetal circulation. This work is expected to serve as a basis for further studies of the diagnosis and treatment of congenital diseases in early pregnancy. PMID- 8414344 TI - A randomized trial of intrapartum electronic fetal heart rate monitoring versus intermittent auscultation. PMID- 8414345 TI - Ultrasound diagnosis of uterine myomas and complications in pregnancy. PMID- 8414346 TI - A 'suitable and sufficient' display screen assessment. AB - Valerie Woods describes a systematic approach to the assessment of display screen work and equipment to enable compliance with recent legislation. PMID- 8414347 TI - Work-related back pain. AB - Sylvia Newman and Cathy Callaghan suggest that much can be done to reduce the incidence of musculoskeletal pain among nursing and midwifery staff following their survey of Brighton Health Care employees. PMID- 8414348 TI - Learning to study the independent way. AB - For those OHPs returning to study at diploma level educational approaches encouraging independent thought can present unexpected new challenges. James Garvey discusses teaching strategies in higher education and argues that it is time to get away from the formal learning situations of the past. PMID- 8414350 TI - AIDS charity calls for nurse education. PMID- 8414349 TI - Crime in the workplace. AB - Witnessing a criminal act in the workplace can place the OHP in a compromising position. Linda Goldman describes the duties of the OHP who is asked to provide evidence of an employee's misdemeanour. PMID- 8414351 TI - AIDS update. Latest WHO estimate. PMID- 8414352 TI - Profile: Graham Johnson. PMID- 8414353 TI - Risk assessments and the law. PMID- 8414355 TI - Unused drug samples. PMID- 8414354 TI - Working in extreme temperatures--cold. PMID- 8414356 TI - Shiftwork and the offshore worker. PMID- 8414357 TI - A clear policy: eyesight testing. AB - Like it or not employers are now required to provide employee eye and eyesight tests on request. Following last month's article on the Display Screen Equipment Regulations Hazel Montague looks at this specific duty and describes how one company has taken a proactive approach to setting up a vision screening programme. PMID- 8414358 TI - Keeping up standards: BS5750. AB - The occupational health department of Dumfries and Galloway Health Board is the first of its kind to gain BS5750. Alison Sloan describes how her department achieved the British Standards Institute's quality assurance accreditation. PMID- 8414359 TI - AIDS update. Nurses ignore procedures. PMID- 8414360 TI - A friend in need or a counsellor? AB - Should workplace counselling be an in-house responsibility or does an external service provide greater benefits? JOHN GOLDMAN weighs up the arguments. PMID- 8414361 TI - NIHL advice: falling on deaf ears? PMID- 8414362 TI - Policing the railways: the unexpected risk. AB - High profile issues--such as the King's Cross disaster--have obscured the daily risks to health encountered by the British Transport Police. ANNETTE CHORLEY describes her efforts to raise HIV and hepatitis B awareness in officers policing the railways. PMID- 8414363 TI - Working in a hot environment. PMID- 8414364 TI - Project 2000 and the working environment. PMID- 8414365 TI - Dermatitis on construction sites. PMID- 8414366 TI - Valuing, empowering employees vital to quality health & safety management. PMID- 8414367 TI - Solving 'white collar' pain problems. PMID- 8414368 TI - Healthcare's environment. PMID- 8414369 TI - Occupational health nurse practitioner has broader legal scope, accessibility. OHNPs receive high ratings for the cost-effective delivery of quality healthcare services. PMID- 8414370 TI - Worksite health promotion bringing companies positive economic impact. PMID- 8414371 TI - National safety congress emphasizes that workplace safety equals quality. PMID- 8414372 TI - Outfitted to filter out respiratory risk. PMID- 8414373 TI - OSHA's respiratory protection standard. PMID- 8414374 TI - Exxon adds respiratory training to modules. PMID- 8414375 TI - Personal protection. PMID- 8414376 TI - Engineering measures can muffle hazards heard in noisy work areas. AB - The OSHA noise standard establishes only minimum levels of protection and exposure. Legally acceptable noise levels may result in a hearing loss among some employees. Noise hazards are best controlled through engineering methods such as enclosures, air exhaust mufflers and vibration damping. If engineering controls do not reduce noise to allowable levels, employee exposure must be controlled via a hearing conservation program that includes provisions for audiometric testing, protective equipment and employee training. PMID- 8414377 TI - A biographical sketch of Henry Hofstetter. AB - This paper describes Dr. Hofstetter's early family life and the influences which shaped his attitudes to his friends, his work, and his profession. It chronicles his early experiences as a young professor of optometry and later as a highly successful administrator. PMID- 8414378 TI - Graduate education in optometry--what do we need? AB - Current needs in graduate education are addressed. In particular, the areas of biological sciences need to be addressed. Mechanisms to achieve a combined degree program are proposed. PMID- 8414379 TI - A history of M.S. and Ph.D. programs offered by schools and colleges of optometry in North America. AB - This paper gives an overview of the history of M.S. and Ph.D. graduate programs offered by schools and colleges of optometry. It also highlights some of the contributions of Henry Hofstetter to graduate education. PMID- 8414380 TI - Who will be tomorrow's optometric educators? PMID- 8414381 TI - Optometric history--who cares? PMID- 8414382 TI - International cooperative exchange programs in clinical optometry. AB - A reciprocal exchange program between the University of Waterloo and the Hong Kong Polytechnic in clinical optometry is presented. Costs and benefits are described. PMID- 8414383 TI - An international perspective on optometric education. AB - In many of the world's nations, optometry hardly exists because resources are not sufficient to educate optometrists nor to fund their services. In others, tradition and accommodation with other forces have rendered optometry incapable of change that would expand its scope of services. But, in a growing number of countries, there is an accelerating trend toward expansion of education and scope of practice. Optometry is coming to be defined in those parts of the globe as that independent primary health profession whose practitioners are educated in vision and health sciences, and who meet standards that qualify them to diagnose and treat visual problems and ocular disease. Review of this change, wherever it has transpired, leads to the conclusion that the scope of optometric practice expands only after corresponding expansion in optometric education. These goals are being achieved in a group of highly developed countries in which optometry has long been a major eye care provider, and in countries in which socio-economic and political conditions are improving, but where there is no significant source or tradition of primary eye care of any scope. PMID- 8414384 TI - Teaching the traditional optometry with the new optometry. AB - This paper addresses the problem of integrating new technology into the preclinical and clinical teaching programs. The use of senior students to teach their juniors is advocated. A team approach to patient care is discussed. PMID- 8414385 TI - The optometric literature: an editor's perspective. AB - I reviewed optometric literature trends and compared them with those of the medical literature. Several journals were surveyed and the published articles classified and counted over 5 or more years. Parallel changes seem to be occurring in the medical and optometric literature, including shorter articles, more authors per paper, and fewer case reports and anecdotal reports. It is impractical and unproductive to attempt to classify a paper as basic science or clinical science because the disciplines overlap. Fads wax and wane over cycles of about 10 years. Clinical research needs more support. PMID- 8414386 TI - The optometric educators' role in public health issues. AB - The optometric educator has a very important role in the development, analysis, interpretation, and promotion of public health issues. Because public health provides the framework for making rational decisions about the complexities of health care, the optometric educator has a high-order responsibility to educate students in the principles, concepts, and practice of public health. As both a creator and a deliverer of knowledge, the optometric educator also must ensure that the outcomes of his scientific inquiry are translated into relevant health policy and used effectively and efficiently for the benefit of all society. Advancement of the public health philosophy within the venue of optometric education should be the responsibility of all faculty, not just those members of the faculty who have direct responsibility for the public health curriculum. PMID- 8414387 TI - 1% Cyclopentolate hydrochloride: another look at the time course of cycloplegia using an objective measure of the accommodative response. AB - The time course of cycloplegia was measured by monitoring residual accommodation after the application of 1 drop (29.3 microliters) of 1% cyclopentolate hydrochloride. Three different measures of residual accommodation were made, one objective assessment with an optometer, and two subjective assessments similar to those used by previous investigators. Pupil diameter was also measured in a subgroup of individuals to compare the time course of the induced mydriasis to that of the cycloplegia. When residual accommodation is measured objectively, maximum cycloplegia occurs 10 min after the application of 1% cyclopentolate hydrochloride in individuals with light irides. This result suggests that the standard clinical protocol of delaying refraction 30 to 60 min after the application of cyclopentolate hydrochloride may be too conservative for individuals with light irides. For individuals with dark irides, 30 to 40 min is required for maximum cycloplegia, and the magnitude of residual accommodation in these individuals is similar to that found in light iris individuals at 10 min. When subjective measures are used to estimate residual accommodation, more accommodation is present and the time at which maximum cycloplegia occurs is delayed for individuals with light irides. These results are in agreement with previous studies using subjective techniques. Regardless of iris color or measurement method, the time course for pupil dilation is not the same as the time course for cycloplegia. PMID- 8414388 TI - Keating's asymmetric dioptric power matrices expressed in terms of sphere, cylinder, axis, and asymmetry. AB - Twelve years ago Keating pointed out that dioptric powers existed which could not be represented by the familiar three parameters sphere, cylinder, and axis. They are the equivalent powers of optical systems (including many eyes) with separated obliquely crossing astigmatic elements. Four parameters are required to represent such powers, and all four are unfamiliar to most clinicians and researchers. This note shows that it is, in fact, possible to transform the four parameters so that the three familiar parameters are retained and only one (called asymmetry) remains unfamiliar. The consequence is that it is always possible to represent a power by means of sphere, cylinder, axis, and asymmetry. Powers commonly used in practice all have asymmetry equal to zero which is why only the first three are usually necessary. Powers, however, do exist, and are of potential interest in optometry, for which asymmetry is not zero and cannot be omitted from the representation. Two numerical examples are given, including Keating's model eye. PMID- 8414389 TI - Normal values of eye position and head size in Chinese children from Hong Kong. AB - We measured the eye position and head dimensions of Chinese children in Hong Kong. Values for exophthalmos, interpupillary distance (IPD), interorbital distance (IOD), distance between medial canthi, and head dimensions were found to be larger than those for Caucasian, Black, or other Chinese groups. PMID- 8414390 TI - Parameter consistency in disposable lenses. AB - An evaluation of parameter consistency and edge defects as they might relate to premature lens fracture was undertaken for 2 disposable lens types, 50 Johnson & Johnson Acuvue lenses and 50 Wesley-Jessen FreshLook lenses. Diameter measurements varied from 13.97 to 14.48 mm (mean = 14.28 mm, SD = 0.138 mm) for Acuvue and 14.30 to 14.63 mm (mean = 14.41 mm, SD = 0.064 mm) for FreshLook. Based on previously reported information, these diameter variations for Acuvue may be sufficient to affect lens fitting characteristics. Calculated back surface sagittal depth varied from 3.27 to 3.38 mm (mean = 3.32, SD = 0.029 mm) for Acuvue and 3.68 to 3.87 mm (mean = 3.75 mm, SD = 0.039 mm) for FreshLook. Center thickness was generally consistent across the power range evaluated (-0.50 to 4.00 D) for the FreshLook lenses, and appeared to increase significantly for low minus powers for the Acuvue lenses. Twenty-one (44%) Acuvue lenses and 6 (12%) FreshLook lenses showed edge defects that might contribute to premature lens fracture. PMID- 8414391 TI - Three-year changes in refraction and its components in youth-onset and early adult-onset myopia. AB - We compared refractive components, and their changes during a 3-year period, for 79 young adults of whom 29 were youth-onset myopes, 26 were early adult-onset myopes, and 24 were emmetropes. In the initial evaluation we found that mean corneal power was greater for both groups of myopes than for the emmetropes, whereas mean vitreous chamber depth and mean axial length were greater for the youth-onset myopes than for the other two refractive error groups. However, the differences between the two groups of myopes appear to be related to the fact that the mean amount of myopia was significantly greater for the youth-onset myopes than for the early adult-onset myopes. During the 3-year period, mean spherical equivalent refraction for subjects in all three refractive error groups changed in the direction of increasing myopia. For each of the three groups, the only refractive component changes having significant correlations with changes in refraction were vitreous chamber depth and axial length. We interpret these results as indicating that: (1) whether axial elongation occurs before, during, or after the completion of the normal growth period, the result is an eye whose cornea is significantly steeper, whose vitreous chamber depth and axial length are significantly greater, and whose lens differs little from that of an emmetropic eye of a person of the same age and (2) when myopia progresses with time, the progression is due to an increase in axial length that is not fully compensated by a decrease in lens power.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414392 TI - Increasing knowledge about the older population: the role of geriatric curriculum in optometry. AB - Education in the area of geriatrics/gerontology alleviates misconceptions about the elderly and lessens ageism. Ageism, negative feelings toward the elderly, represents a substantial obstacle to the delivery of quality health care to older adults. As future primary health care professionals, optometry students will encounter a large segment of the older population. This study looks at the effect that a didactic course and a clinical rotation in geriatrics have on the knowledge and attitude of optometry students toward older adults. Palmore's Facts on Aging Quiz: Part 1 was administered to two classes of optometry students. One class experienced a geriatric course and clinic concurrently; the other class experienced the geriatric clinic before the course. Scores increased significantly post-test for both classes. Exposure to geriatric patients before didactic instruction did not improve pretest scores; however, the number of patient exposures was significant at post-test. Antiage bias scores also improved with intervention of a didactic course and clinical rotation in geriatrics. The results of this study indicate that ageism in optometrists may be ameliorated by providing formal instruction in geriatrics in the optometric curriculum. PMID- 8414393 TI - Vernier acuity through night vision goggles. AB - Night vision goggles (NVG's) are being used increasingly in military and civilian environments. Despite the use of these devices, relatively few tests exist to assess visual performance through NVG's. Hyperacuity tasks may provide a sensitive index of performance through night vision devices. In this study, grating vernier acuity was measured through NVG's. As reported previously, a power law relation was observed between vernier acuity and stimulus contrast. Comparison of vernier acuity with and without NVG's indicated that performance is limited by the contrast transfer of the device. Vernier acuity measurements can be used to assess the quality of vision and quantity of contrast transferred through night vision devices. PMID- 8414394 TI - Mechanisms of accommodation. PMID- 8414395 TI - Cryotherapy of conjunctival melanoma. PMID- 8414396 TI - Criteria for success of glaucoma surgery. PMID- 8414397 TI - A-scan in Graves ophthalmopathy. PMID- 8414398 TI - Globe perforation during anesthetic injection. PMID- 8414399 TI - Complications of optic nerve sheath decompression. PMID- 8414400 TI - The sensitive period for strabismic amblyopia. PMID- 8414401 TI - Incentives for healthcare screening. PMID- 8414402 TI - A randomized trial of intraocular lens fixation techniques with penetrating keratoplasty. AB - PURPOSE: Pseudophakic corneal edema is the principal indication for penetrating keratoplasty in the United States. Currently, three techniques of intraocular lens (IOL) fixation during penetrating keratoplasty for this condition are commonly used--flexible anterior chamber IOL (AC IOL) implantation, iris suture fixation of a posterior chamber IOL (PC IOL), and transscleral suture fixation of a PC IOL. This study represents the first prospective, randomized comparison of these three techniques. METHODS: One hundred seventy-six consecutive patients with pseudophakic corneal edema who underwent penetrating keratoplasty with IOL exchange were randomized to one of the three implantation techniques. Standardized evaluations were performed at baseline and at 6, 12, and 18 months postoperatively. Life-table analysis provided cumulative risk estimates for specific complications. RESULTS: Randomization produced comparable groups at baseline. The cumulative risk of macular edema was significantly less for the iris fixation cohort than for either the AC IOL or scleral fixation group. A complications index was constructed based on the major adverse outcomes of glaucoma escalation, cystoid macular edema, IOL dislocation, and graft failure. A significantly lower risk of complication was found for iris compared with scleral fixation of PC IOLs. CONCLUSION: The authors conclude that transscleral fixation of the PC IOL at the time of penetrating keratoplasty for pseudophakic corneal edema is associated with a greater risk of adverse outcome than iris fixation of a PC IOL. PMID- 8414403 TI - Clinical and diagnostic use of in vivo confocal microscopy in patients with corneal disease. AB - BACKGROUND: The purpose of this article is to introduce the practicing ophthalmologist to the optical principles and images produced by a tandem scanning confocal microscope (recently approved by the Food and Drug Administration for general clinical use). The tandem scanning confocal microscope allows real-time viewing of structures in the living cornea at the cellular level in four dimensions (x, y, z, and time). METHODS: Nine patients (2 males, 7 females), ranging in age from 7 to 52 years, were examined. Images were recorded on super VHS videotape, digitized and processed on a computer workstation, and photographed for presentation. RESULTS: Two-dimensional (x, y) 400 x 400-microns images (9-microns z-axis thickness) are presented for normal corneal structures and for the clinical conditions of herpetic keratitis, wound healing after myopic excimer ablation, Acanthamoeba infection, corneal dystrophies (granular, Reis Buckler), contact lens abrasion, and the irido-corneal endothelial syndrome. CONCLUSION: Clinical confocal microscopy has the unique potential of providing noninvasive assessment of corneal injury and disease at the cellular level that is not available currently from other technologies. PMID- 8414404 TI - Spheroidal keratopathy associated with subepithelial corneal amyloidosis. A clinicopathologic case report and a proposed new classification for spheroidal keratopathy. AB - PURPOSE: An unusual case of primary spheroidal degeneration associated with subepithelial corneal amyloidosis is described clinically and histopathologically, and a new classification for spheroidal keratopathy is proposed. METHODS: The corneal button obtained by lamellar keratoplasty was examined by light and transmission electron microscopy. Amyloid deposits were evaluated further by means of the thioflavin T fluorescence technique, the potassium permanganate reaction, and immunohistochemical studies using antibodies against immunoglobulin light chains and proteins AA and AP. RESULTS: Spheroidal droplets were observed in basement membrane, Bowman's layer, and anterior stroma, and exhibited characteristic staining with Verhoeff's iron hematoxylin. Amyloid deposits were found beneath the epithelium, almost exclusively in the central area of the corneal button. The deposits showed no immunohistochemical staining with antibodies to amyloid proteins AA and AP or kappa and lambda light chains. CONCLUSIONS: The histochemical properties of the spheroidal droplets in this case were similar to those observed by other authors and suggested a sulfur-rich protein other than keratin. The combination of clinical and pathologic findings permitted the differential diagnosis of subepithelial amyloidosis and distinguished this condition from that of gelatinous drop-like dystrophy. To the authors' knowledge, this is the first description of primary spheroidal keratopathy associated with secondary subepithelial corneal amyloidosis in an eye that had not undergone previous keratoplasty. PMID- 8414405 TI - Ocular and ocular adnexal injuries treated by United States military ophthalmologists during Operations Desert Shield and Desert Storm. AB - BACKGROUND: Ocular and ocular adnexal injuries, both combat-related and accidental, are common during wartime. In a combat setting, the eye is particularly vulnerable to serious injury from tiny flying particles that might minimally affect other parts of the body. The purpose of this study is to examine the incidence of serious ocular and ocular adnexal injuries that occurred during Operations Desert Shield and Desert Storm. METHODS: The authors retrospectively reviewed serious ocular and ocular adnexal injuries treated by United States Army and Navy ophthalmologists that occurred during Operations Desert Shield and Desert Storm. Only those injuries that resulted in, or would have resulted in, hospital admission because of the ocular or ocular adnexal injury alone are presented. RESULTS: During Desert Shield, 20 patients (23 eyes) suffered serious ocular or ocular adnexal injuries compared with 160 patients (198 eyes) in Desert Storm. During Desert Storm, 78% of all serious injuries were caused by blast fragmentation from munitions. More than one third of the 98 globe lacerations reported in this article were 10 mm or less in size. Of 35 enucleations performed during Desert Storm, 94% were the result of munitions fragments. CONCLUSIONS: During Operation Desert Storm, fragmentation wounds from munitions were the most common cause of ocular and ocular adnexal morbidity. The authors' findings indicate that polycarbonate ballistic protective eyewear could have prevented many of the ocular injuries that they report. PMID- 8414406 TI - Infectious endophthalmitis after penetrating injuries with retained intraocular foreign bodies. National Eye Trauma System. AB - PURPOSE: To determine the risk factors and prognostic indicators of infectious endophthalmitis in eyes with penetrating injury and retained intraocular foreign body. METHODS: From the National Eye Trauma System (NETS) Registry, 492 eyes with intraocular foreign bodies were reviewed for signs of infectious endophthalmitis. RESULTS: Thirty-four eyes (6.9%) with intraocular foreign bodies had evidence of infectious endophthalmitis, and 31 (91.2%) of those eyes had signs of infection at the time of removal of the intraocular foreign body. The majority of eyes with an intraocular foreign body with or without endophthalmitis were in patients between 10 and 39 years of age, but the risk of endophthalmitis developing increased with age, especially in patients 50 years of age or older with delayed primary repair (P = 0.005). Endophthalmitis was more likely to develop in eyes with home or occupational injuries (33/358, 9.2%) than in those with injuries from other settings (1/128, 0.8%; P = 0.001). Infectious endophthalmitis was much less likely to develop in eyes with primary repair within 24 hours of the injury (10/287 = 3.5%) than in eyes with primary repair more than 24 hours after the injury (22/164, 13.4%; P < 0.0001). Bacilli or staphylococci were isolated in 21 (95%) of 22 eyes with positive cultures. Visual prognosis was reasonably good with 15 (58%) of 26 eyes attaining a visual acuity of 20/200 or better. CONCLUSIONS: Removal of a retained intraocular foreign body within 24 hours of injury markedly reduces the risk of infectious endophthalmitis developing. Older persons are at high risk for endophthalmitis developing after retaining an intraocular foreign body when there is delayed surgical repair. PMID- 8414407 TI - Hydraulic orbital injection injuries. AB - BACKGROUND: A high-pressure stream from mechanical equipment may inject gas or liquids deep into the orbit with few initial clinical signs. Aggressive surgical debridement as used in the extremities for the treatment of injection injuries is not possible in the orbit. METHODS: Four patients with orbital injection injuries from farm or industrial equipment are presented. Previously reported cases of high-pressure injection injury are reviewed. RESULTS: Two patients suffered localized anterior orbital inflammation partially responsive to steroidal and nonsteroidal anti-inflammatory agents. Late debridement was required in one patient for a persistent lipogranuloma. Two patients suffered more dramatic and diffuse injections of hydrocarbon mixtures, requiring emergent early surgical debridement and decompression for compressive orbital signs. All patients attained an adequate functional outcome, with one patient's vision limited by a coexisting ocular injury. CONCLUSIONS: High-pressure orbital injection injuries manifest a spectrum of signs ranging from acute inflammation with tissue necrosis and compressive visual loss to late chronic inflammation with a pseudotumor-like course. The authors recommend the initial treatment of orbital injection injuries with systemic antibiotics followed by prompt neuroradiologic imaging. Systemic corticosteroids should be added for confirmed injection injuries with surgical debridement of discrete masses and orbital decompression when indicated. Continued therapy with anti-inflammatory medication may be required to suppress chronic inflammation with selective late surgical debridement of lipogranulomas. PMID- 8414408 TI - The prevalence of neurologic dysfunction in children with strabismus who have superior oblique overaction. AB - BACKGROUND: Children with certain neurologic diseases (hydrocephalus, meningomyelocele, or cerebral palsy) have been reported to manifest a high frequency of A-pattern strabismus and superior oblique overaction. However, it is not generally recognized whether children with strabismus who have superior oblique overaction are more likely to have concurrent neurologic diseases than those without superior oblique overaction. In this study, the authors examine this issue. METHODS: The authors retrospectively reviewed the medical records of all patients (n = 168) with overdepression of the downturned eye in adduction, who were examined between October 1989 and March 1992. A randomly selected population of children with strabismus who did not have overdepression of the eye on infraduction and adduction served as controls (n = 98). Patients with simulating or confounding conditions such as pseudo-superior oblique overaction, inferior rectus skew deviation (alternating skew on lateral gaze), and restrictive or paralytic strabismus, and who were older than 20 years of age, were excluded. RESULTS: One hundred twelve patients with true superior oblique overaction were analyzed. Of these 112 patients, 45 (40.2%) had concurrent neurologic abnormalities, compared with less than one fifth (17.3%) of control subjects (17 of 98) (P < or = 0.001). CONCLUSIONS: Children with strabismus who have superior oblique overaction were found to have higher prevalence of concurrent neurologic diseases than control subjects. Superior oblique overaction may represent a clinical marker for an associated neurologic dysfunction, possibly representing a form of skew deviation in some cases. PMID- 8414409 TI - Improvement of visual acuity after surgery for nystagmus. AB - PURPOSE: The authors compared the preoperative and postoperative binocular visual acuities and eye movement recordings of patients who underwent eye muscle surgery consisting of the Anderson-Kestenbaum procedure or the artificial divergence procedure modeled after Cuppers, or a combination of both procedures, for the treatment of infantile nystagmus with head turn. METHODS: Binocular visual acuities and eye movement recordings by electro-oculography (EOG) were compared preoperatively with those done within 3 weeks postoperatively. Shifting and broadening of the minimal intensity zone and foveation time and changing of the waveform were measured. The treatment of 6 of 18 patients was based on the artificial divergence principle; for 7 patients, treatment was in accordance with the Anderson-Kestenbaum principle; and 5 patients had combined procedures. RESULTS: The improvement in binocular visual acuities was two Snellen lines or more in one of six patients in the artificial divergence group and four of five patients in the combined treatment group. The EOG recordings showed shifting of the minimal intensity zone toward the primary position in all three groups. A broadening of the minimal intensity zone occurred mostly in the artificial divergence and combined groups. Increases in foveation time and changes in waveforms from jerk to jerk with foveation were found in three of six patients in the artificial divergence group and in two of five patients in the combined group. CONCLUSION: With less muscle surgery, the artificial divergence and combined operations gave better vision improvement than the Anderson-Kestenbaum operation. PMID- 8414410 TI - Comparisons of methods to detect glaucomatous optic nerve damage. AB - PURPOSE: Various techniques of optic disc and nerve fiber layer evaluation may be used to detect structural glaucomatous damage. The authors compared several qualitative and quantitative methods to determine their relative sensitivities and specificities to detect the presence of glaucomatous visual field loss. METHODS: Fifty-one healthy eyes, 169 ocular hypertensive eyes with normal visual fields, and 132 glaucomatous eyes with early visual field defects were evaluated with qualitative and quantitative measures of structural damage to the optic nerve and nerve fiber layer. Qualitative evaluations were performed by three experienced masked observers who independently graded stereoscopic color disc and monochromatic nerve fiber layer photographs. Quantitative measurements of disc rim area and nerve fiber layer height were made with digitized image analysis of videographic images. Manual planimetric measurements of disc rim area were made from enlarged prints of stereoscopic optic disc photographs. Diagnostic precision was defined as the total proportion of correct diagnoses for the presence or absence of visual field loss. RESULTS: The diagnostic precision of results of a quantitative disc examination (81%) was greater than those of a qualitative nerve fiber layer examination (75%). Quantitative nerve fiber height measurement had the highest sensitivity rate (73%) and results of the qualitative disc examination had the highest specificity rate (87%) of the methods tested. CONCLUSION: The diagnostic precision of disc evaluation was superior to other methods, including nerve fiber layer examination, in correctly determining the presence of structural glaucomatous damage at the early visual field loss stage. PMID- 8414411 TI - The diagnosis of diabetic retinopathy. Ophthalmoscopy versus fundus photography. AB - PURPOSE: To compare fundus photography with ophthalmoscopy in the detection of diabetic retinopathy. METHODS: Ophthalmoscopy and fundus photographs with a nonmydriatic camera, both performed through dilated pupils, were compared to diagnose retinopathy in a cohort of 410 Oklahoma Indians with noninsulin dependent diabetes mellitus. A total of 795 eyes were examined using both methods. The mean age of participants was 60.3 years, with a mean duration of diabetes of 17.3 years. RESULTS: An overall agreement of 86.3% with a kappa statistic kappa of 0.74 was found between ophthalmoscopy and fundus photography with a nonmydriatic camera. For the diagnosis of proliferative diabetic retinopathy, kappa = 0.84 with an agreement of 98.1%. With a total of 61 cases of proliferative retinopathy diagnosed by either method in our study, ophthalmoscopy alone detected 88.5% and fundus photography, 78.7%. When compared on a lesion-by lesion basis, agreement between the two diagnostic methods was highest for nonproliferative retinopathy, as well as fibrous proliferation. CONCLUSION: The fundus photography with a nonmydriatic camera, performed with mydriasis, is comparable to ophthalmoscopy for the detection of retinopathy. It may prove to be a suitable, cost-effective method for routine screening in diabetes clinics, provided ophthalmologic referral is ensured for those with a diagnosis of any form of retinopathy, questionable retinopathy, nondiabetic retinopathy, those with poor quality photographs, as well as those with acute changes in visual acuity. PMID- 8414412 TI - Assessment of vision in idiopathic macular holes with macular microperimetry using the scanning laser ophthalmoscope. AB - BACKGROUND: Visual loss in eyes with full-thickness macular holes has been thought to be due to the absence of retinal function in the area of neurosensory defect as well as loss or reduction of retinal function in the surrounding area of neurosensory retinal detachment. With the advent of surgical techniques to treat macular holes, it is increasingly important to better characterize this visual dysfunction. METHODS: Thirty eyes of 30 patients with full-thickness idiopathic macular holes were evaluated with microperimetry using the scanning laser ophthalmoscope to detect and quantitate absolute and relative scotomata within the central 40 degrees of visual field. A log 2 scale of test stimulus intensities was established. Results of microperimetry were compared with best corrected visual acuities as measured on the logarithmic Early Treatment of Diabetic Retinopathy Study chart as well as duration of symptoms. RESULTS: All 30 eyes showed an absolute scotoma in the area of neurosensory defect as well as surrounding relative scotomata in the area of neurosensory detachment. Best corrected visual acuity was correlated with the size of the absolute and relative scotomata (P < 0.002). The sizes of the scotomata were correlated with the duration of symptoms of the macular holes (P < 0.05). CONCLUSION: Microperimetry using the scanning laser ophthalmoscope demonstrates that the visual loss associated with macular holes is related to the reduction of retinal function in the area of the surrounding neurosensory detachment as well as the absence of retinal function in the area of neurosensory defect. The size of the scotomata, determined by microperimetry, is correlated with the patient's visual acuity as well as the duration of symptoms of the macular hole. PMID- 8414413 TI - Idiopathic juxtafoveolar retinal telangiectasis. Update of classification and follow-up study. AB - BACKGROUND: Idiopathic juxtafoveolar retinal telangiectasis may cause visual loss. The treatment of this disease is controversial. METHODS: The authors reviewed the records of 140 patients with idiopathic juxtafoveolar telangiectasis. A classification scheme based on biomicroscopic and fluorescein angiographic findings is presented. In addition, the effect of photocoagulation on the natural history of the disorder is evaluated. FINDINGS: Patients are categorized into three groups. Group 1 comprises 39 male patients with nonfamilial, easily visible telangiectasis and intraretinal exudation. The telangiectasis is unilateral in 94% of patients. The telangiectasia in this group is probably of developmental origin (Coats syndrome). Group 2 comprises 94 patients with occult juxtafoveolar telangiectasis, minimal exudation, superficial retinal crystalline deposits, and right-angle venules. Late in the course of the disease, foveolar atrophy, intraretinal pigment plaques, and subretinal neovascularization develop. The telangiectasis is acquired during middle age and is bilateral in 98% of patients. Group 3 comprises seven patients with bilateral easily visible telangiectasis, minimal exudation, and capillary occlusion. All of these patients had systemic disease, which was probably related to their eye disease. CONCLUSION: Slow visual loss beginning in adulthood characterizes most of these patients. The telangiectasis appears to be caused primarily by retinal capillary leakage in group 1, capillary diffusion abnormalities in group 2, and capillary occlusion in group 3. Photocoagulation is probably beneficial for patients in group 1 and not for patients in group 2, at least before their development of subretinal neovascularization. PMID- 8414414 TI - Helium ions versus iodine 125 brachytherapy in the management of uveal melanoma. A prospective, randomized, dynamically balanced trial. AB - PURPOSE: Optimal radiation therapy for uveal melanoma is uncertain, and the relative efficacies of radioactive plaques and charged particles are unclear. METHODS: The authors prospectively studied helium-ion irradiation and iodine 125 (125I) brachytherapy in a randomized, dynamically balanced trial. Of the 184 patients who met the eligibility criteria, 86 were treated with helium ions and 98 with 125I brachytherapy. RESULTS: No patients with uveal melanoma had a history of systemic malignancy. Tumors were less than 15 mm in maximum diameter and less than 10 mm in thickness. A minimum tumor dose of 70 GyE was delivered to the tumor apex. There was a significantly higher local recurrence rate after 125I brachytherapy than after helium-ion irradiation. Enucleations occurred more frequently after brachytherapy (relative risk = 1.99; 95% confidence interval, 0.78-5.78). More anterior segment complications occurred after helium-ion irradiation. To date, there has been no measurable impact on survival. CONCLUSIONS: Most uveal melanomas can be managed with radiation with retention of the eye. There was better tumor control with helium-ion irradiation; however, there were more anterior segment complications. PMID- 8414415 TI - The time course of irradiation changes in proton beam-treated uveal melanomas. AB - BACKGROUND: Although histopathologic changes in uveal melanomas treated by irradiation have been well characterized, the timing of these effects after irradiation has not been previously evaluated. METHODS: The series consisted of a total of 92 eyes with uveal melanoma, enucleated at various intervals after proton irradiation (range, 1 month to 8 years) due to complications (n = 66) or tumor growth (n = 22), or at autopsy (n = 4). Study slides were read, using a standard protocol, by two pathologists masked to the timing of the enucleation after irradiation and the reason for the surgery. RESULTS: The prevalence of inflammation decreased whereas fibrosis increased with time from 15% of cases enucleated within 12 months (early cases) to 61% of those enucleated more than 30 months (late cases) after irradiation (P for trend = 0.0005). Tumor necrosis and blood vessel damage occurred early, and the prevalence of these changes was constant over time. Excluding tumors with evidence of growth after irradiation, mitotic figures became progressively less common as the interval between irradiation and enucleation increased (P for trend = 0.015); no mitotic figures were present in 40 high-power fields more than 30 months after irradiation. CONCLUSION: Histopathologic changes in irradiated melanomas vary according to the time elapsed since irradiation. Inflammation decreases with time whereas fibrosis becomes more prevalent with time after irradiation. Among controlled tumors, mitotic figures appear only in recently irradiated tumors. PMID- 8414416 TI - Retinal hemodynamics using scanning laser ophthalmoscopy and hemorheology in chronic open-angle glaucoma. AB - PURPOSE: Recent studies suggest that elevated intraocular pressure is not the only causative factor for the development of visual field loss and optic nerve damage in glaucomatous eyes. The authors determine whether retinal hemodynamics or blood fluidity are alternated in eyes of patients with open-angle glaucoma compared with those of age- and sex-matched healthy subjects. METHOD: High quality video fluorescein angiograms were obtained from single eyes of 51 patients with chronic open-angle glaucoma. From these angiograms, the arm-retina time, mean dye velocity, and arteriovenous passage time were quantified. The data from patients were compared with those of an age- and sex-matched group of healthy subjects. RESULTS: In patients with chronic open-angle glaucoma, an 11% reduction of the mean dye velocity (P < 0.05) and a 41% prolongation of the arteriovenous passage time (P < 0.01) was observed relative to the values obtained among the control subjects. Among hematocrit values, plasma viscosity, and erythrocyte aggregation, only plasma viscosity showed a significant increase (4%; P < 0.01) in patients with chronic open-angle glaucoma. CONCLUSION: These results indicate that a pronounced circulatory deficit exists within the retinal vasculature of glaucomatous eyes, which may coexist with, but cannot be fully attributed to, an increase in plasma viscosity. PMID- 8414417 TI - Diode laser surgery. Ab interno and ab externo versus conventional surgery in rabbits. AB - BACKGROUND: Fibroblastic proliferation of subconjunctival tissues remains a primary mechanism of failure in filtration surgery. Minimizing the surgical manipulation of episcleral tissues may reduce scarring. Laser sclerostomy surgery involves minimal tissue dissection, and is gaining attention as a method of potentially improving filter duration in high-risk cases. METHODS: Twenty-five New Zealand rabbits underwent filtration surgery in one eye, and the fellow eye remained as the unoperated control. Ten rabbits underwent ab externo diode laser sclerostomy surgery, ten underwent ab interno diode sclerostomy surgery, and five had posterior sclerostomy procedures. Filtration failure was defined as a less than-4-mmHg intraocular pressure (IOP) difference between the operative and control eyes. RESULTS: The mean time to failure for the ab externo, ab interno, and conventional posterior sclerostomy techniques measured 17.4 +/- 11.5, 13.1 +/ 6.7, and 6.0 +/- 3.1 days, respectively. In a comparison of the laser-treated groups with the conventional procedure, the time to failure was significantly longer (P = 0.02) for the ab externo filter. The mean ab interno sclerostomy duration was longer than the posterior lip procedure, but this difference was not statistically significant (P = 0.15). The overall level of IOP reduction was similar in the three groups. CONCLUSIONS: These data suggest that diode laser sclerostomy is a feasible technique in rabbits, and the ab externo approach resulted in longer filter duration than the conventional posterior lip procedure in this model. PMID- 8414418 TI - Aqueous and vitreous concentration of mitomycin C by topical administration after glaucoma filtration surgery in rabbits. AB - PURPOSE: The authors investigated the aqueous and vitreous pharmacokinetics of mitomycin after postoperative topical administration in rabbits. METHODS: Filtration surgery was performed in one eye of each rabbit. On the first postoperative day, mitomycin solution (0.4 mg/ml) was administered by either topical drop or cellulose sponge in both eyes of each rabbit. Aqueous and vitreous paracenteses were performed at 0.25, 0.5, 1, 2, 3, and 4 hours thereafter. Three rabbits were tested at each time interval for each type of administration. RESULTS: After topical drop administration, the peak aqueous concentration was 0.03 +/- 0.02 microgram/ml (mean +/- standard error) in surgical eyes and 0.06 +/- 0.03 microgram/ml in control eyes. After sponge administration, the peak aqueous concentration was 0.10 +/- 0.03 microgram/ml in surgical eyes and 0.08 +/- 0.04 microgram/ml in control eyes. Peak aqueous concentrations from drop and sponge applications were achieved at 1 to 2 hours after administration. Vitreous levels were well below the concentration known to cause retinal toxicity at all time intervals tested. CONCLUSION: Postoperative topical administration of mitomycin was successful in delivering mitomycin into the aqueous humor of rabbit eyes. Alternative methods of mitomycin application from the currently popular intraoperative administration may be beneficial in situations in which mitomycin delivery across intact conjunctiva may be desirable. PMID- 8414419 TI - An association between hypothyroidism and primary open-angle glaucoma. AB - PURPOSE: To test the hypothesis that there is an association between hypothyroidism and primary open-angle glaucoma. METHODS: The study was conducted in a case-control fashion. Sixty-four patients with primary open-angle glaucoma were evaluated for hypothyroidism by history and by undergoing a thyroid stimulating hormone immunoradiometric assay. Sixty-four control subjects from the general eye clinic were evaluated in the same manner. Patients found to have elevated thyroid-stimulating hormone immunoradiometric assay were evaluated by an endocrinologist for hypothyroidism. RESULTS: Of the primary open-angle glaucoma group, 23.4% had hypothyroidism. A diagnosis was made previously in 12.5% patients, and 10.9% were newly diagnosed. Of the control subjects, 4.7% had hypothyroidism. A diagnosis had been made previously in 1.6% of the control subjects, and 3.1% were newly diagnosed. The difference between the two groups was found to be statistically significant. CONCLUSION: A statistically significant association between hypothyroidism and primary open-angle glaucoma is demonstrated. There is a large group (10.9%) of patients with primary open-angle glaucoma with undiagnosed hypothyroidism. PMID- 8414420 TI - Prospective study of sub-Tenon's versus retrobulbar anesthesia for inpatient and day-surgery trabeculectomy. AB - PURPOSE: Several retrospective studies have demonstrated the safety and efficacy of sub-Tenon's anesthesia in ocular surgery. This is the first prospective randomized study comparing sub-Tenon's versus retrobulbar anesthesia for glaucoma surgery. METHODS: Thirty-nine patients undergoing both inpatient and day surgery trabeculectomy were randomized to receive retrobulbar or sub-Tenon's anesthesia. Retrobulbar anesthesia consisted of a 1.5-ml injection of a 1:1 mixture of 2% lidocaine without epinephrine and 0.5% plain bupivacaine with hyaluronidase. Sub Tenon's anesthesia consisted of 2% lidocaine without epinephrine injected over the superior, medial and lateral recti muscles. Both groups received a van Lint lid block and a standardized sedative. Outcome parameters evaluated included patient demographics, operative complications, intraoperative and postoperative patient comfort, and volume of anesthetic. RESULTS: Seventeen patients were randomized to the retrobulbar group and 22 to the sub-Tenon's group. Patient discomfort was statistically similar between the groups. There was no statistical difference between groups with respect to quantity of sedative received, surgical exposure, eye movements, or operative complications. A retrobulbar hemorrhage, however, developed at the time of retrobulbar anesthesia in one patient, requiring cancellation of surgery. A larger volume of local anesthetic was required in the retrobulbar group versus the sub-Tenon's group (1.8 versus 1.1 ml; P < 0.01). Patients receiving retrobulbar injections were more likely to require additional anesthesia (P < 0.01) and postoperative analgesics (P < 0.05) compared with patients undergoing sub-Tenon's injection. There was no significant difference between the groups with respect to age, sex, or operated eye. CONCLUSIONS: Sub-Tenon's anesthesia is safe and effective for patients undergoing either inpatient or day-surgery trabeculectomies, and it requires less local anesthetic than retrobulbar anesthesia. PMID- 8414421 TI - Malignant acanthosis nigricans. A case report. AB - PURPOSE: The authors present a case of malignant acanthosis nigricans that has two unusual aspects. The first is involvement of the lid margins with confluent papillomata, causing severe visual impairment, and the second is the association with lung malignancy. METHODS: A 65-year-old Portuguese man presented with decreased vision, papillomatous lid lesions, pruritus, and progressive thickening of the skin of his face, neck, axillae, and inguinal creases. A diagnosis of acanthosis nigricans was made after skin biopsy. Systemic investigations showed a suprahilar mass, and biopsy was positive for squamous cell carcinoma of the lung. RESULTS: The patient underwent excision of papillomata from all four lid margins, and this was repeated 1 year later. He also completed a course of palliative radiotherapy. CONCLUSION: Malignant acanthosis nigricans is rarely associated with lung neoplasms, being more commonly a manifestation of intra-abdominal malignancies. Of note in this case is the extensive ocular involvement and visual impairment. PMID- 8414422 TI - The response to particulate debris. AB - The most common cause of failure of total hip and other arthroplasties is aseptic loosening of the prosthesis. Substantial evidence suggests that the adverse tissue response to prosthesis wear particles is an important contributor to bone loss around implants. The results may be an increased risk of loosening and severe bone loss, which makes revision surgery more difficult. Studies in humans have demonstrated that the appearances of the periprosthetic tissues are related to the type, number, and size of wear particles. The appearance and tissue response around any given prosthesis is related to the balance among the rate of production of wear particles, the ability of the tissues to deal with the particles, and the rate of clearance of the particles from the joint. Prosthesis wear particles may be produced by wear at the articulating surface, particularly if there is three-body wear, by abrasion due to movement at the prosthesis-bone interface of uncemented prostheses, by abrasion at the prosthesis-cement and cement-bone interfaces of cemented prostheses, or at any articulation of the components of modular prostheses. Future in vivo and in vitro studies of the effects of wear particles should focus on quantitative measurement of the effects of particles and on the use of particles of comparable size and in concentrations similar to those found in the tissues surrounding failed prostheses. PMID- 8414423 TI - Mechanisms of bone loss associated with total hip replacement. AB - The pathogenesis of bone loss after total hip arthroplasty is discussed. Two separate processes are considered: adaptive bone remodeling ("stress-shielding") and osteolysis. Clinical and experimental studies that provide the basis for current and future strategies to minimize the occurrence and severity of these phenomena are summarized. PMID- 8414424 TI - Cementless versus cemented total hip arthroplasty. A scientific controversy? AB - The controversy surrounding cementless versus cemented total hip arthroplasty cannot be resolved at this stage on a strictly scientific basis. Various aspects of this controversy are presented. The focus of some specific discussion sections include acrylic cement, component design of both the socket and the stem, and function and wear. PMID- 8414425 TI - The rationale for cemented total hip arthroplasty. AB - Long-term follow-up of early Charnley cemented THAs demonstrates excellent survival (Table 1). New techniques in cementing have improved the quality of the femoral cement fixation and have shown consistently good performance at 10-year follow-up. Noncemented designs have not yet demonstrated similar long-term results. Recent reports reveal osteolysis in noncemented prostheses, which is observed earlier than in cemented designs. This implies a greater role for polyethylene debris in the etiology of osteolysis. Polyethylene debris is produced not only at the articulation but also at the nonarticular metal polyethylene interface in modular metal-backed acetabular components. This results in the production of additional plastic debris. Metal backing has not demonstrated any clinical improvement in the long-term performance of cemented acetabular components. Furthermore, it may be detrimental due to decreased polyethylene thickness, increased stress within the polyethylene, and an increased rate of polyethylene wear of both the articular and nonarticular surfaces. Multiple reports have demonstrated that titanium alloy is not an acceptable articulating surface because it has a poor resistance to abrasion. It can result in severe metallosis in the periarticular tissues, leading to progressive osteolysis and early failure of the arthroplasty. Caution is suggested in the widespread application of polymodular femoral components because the production of metallic debris may prove excessive. Cemented THA remains the gold standard by which other methods of fixation must be assessed. The enviable long-term results with early cementing techniques and the Charnley prosthesis will be difficult to match, even with developing technology. PMID- 8414426 TI - Total hip replacement in the middle-aged patient. Contemporary cementing for fixation of the femoral component. AB - Important advances have been made in the preparation and use of cement and the design of femoral components. Additionally, we have a greater understanding of the biologic basis of osteolysis, previously considered a manifestation of 'cement disease.' The results now emerging as an outcome of modern cementing techniques and component designs represent a major step forward and raise serious doubts about the justification for cementless fixation of the femoral component, given the present state of the art. PMID- 8414427 TI - The rationale for cementless total hip replacement. AB - Current cementing techniques of distal plugging, pulsatile water cleaning of the canal, and retrograde filling with PMMA in a low-viscous state prepared using porosity reduction techniques and then pressurized have been shown to give excellent results at 10 or more years after operation. However, physical and aging characteristics of PMMA do not guarantee that those results will hold up in the 20- to 30-year time frame. Fifteen-year experience with bony ingrowth systems indicate the development of a durable interface without PMMA, although the interface does appear more vulnerable to attack by the biologic reaction to HMWPE debris. Bone remodeling appears more favorable around proximally fixed bone ingrowth prostheses than around distally fixed prostheses. Softening of the physical characteristics of the stem tip may reduce the incidence of thigh pain. Patients with high activity potential and a life expectancy of greater than 25 years should be strongly considered for a proximally fixed bone ingrowth or ongrowth prosthesis. PMID- 8414428 TI - The rationale for cementless revision total hip replacement with contemporary technology. AB - Undoubtedly, the number of revisions for failed cemented total hip arthroplasties will increase over time based on current survival statistics. Although contemporary cementing techniques may improve the outcome of some procedures, long-term results of cemented revisions are less than ideal. Poor bone quality with cortical deficiencies and endosteal sclerosis make long-term stability and implant survival less likely with cemented techniques. In the very elderly patient requiring revision, PMMA may provide a simpler option with an adequate short-term result. Using uncemented techniques, the complications associated with the inherent properties of cement can be avoided. Bone grafting is a viable option, with the hope that long-term implant stability can be achieved by biological ingrowth into porous surfaces. Reconstitution of previously attenuated cortical bone is seen after bone grafting in association with a noncemented femoral or acetabular implant, although ingrowth into porous-coated surfaces via allograft bone is unlikely. In general, short- and mid-term results with respect to clinical performance are good or excellent in at least 90% of the cases evaluated. With the increased application of uncemented components to revision situations, attention may shift from "cement" disease to "polyethylene" disease. Although not the focus of this article, recent literature with respect to primary uncemented hip and knee arthroplasty supports this contention. We believe that proximal canal fill and biologic ingrowth can provide implant stability, avoid proximal stress shielding of the femur, and improve long-term implant survival and clinical success in the revision situation. PMID- 8414429 TI - Revision total hip arthroplasty. Long-term results without cement. AB - The authors present their results for cementless femoral revision arthroplasty using extensively porous coated implants. Patients have been prospectively followed for 5 to 11 years (mean 7.4 years). The revision rate was 5.7%, and the radiographic loosening rate was 1.1%, to give a mechanical failure rate of only 6.9%. As a result of the revision procedure, the pain level was improved for 89.1% of the patients, walking status was improved for 82.8% of the patients, and functional level was improved for 88.5% of the patients. These results compare very favorably with any reports on revision arthroplasty using cement with a similar length follow-up. PMID- 8414430 TI - Thigh pain in cementless total hip arthroplasty. A comparison of two systems at 2 years' follow-up. AB - Cementless total hip arthroplasties using 110 porous-coated anatomic (PCA) and 105 consecutive Mallory-Head (MH) prostheses were reviewed using prospectively gathered clinical data and a detailed radiographic analysis. In an earlier experience with the PCA, 13% of patients experienced thigh pain at 1 year and 23% at 2 years. Positive radiographic correlates with thigh pain included subsidence, distal cortical hypertrophy, a tight diaphyseal fit, and stem tip sclerosis. In the current MH study group, the incidence of thigh pain was 7% at 1 year and 3% at 2 years. No radiographic findings correlated positively with thigh pain. The results of this short-term clinical comparison suggest that a long, straight, tapered design offered better initial stability than its anatomic counterpart as reflected in the much lower incidence of thigh pain. These results may translate into improved osseous integration and long-term success. PMID- 8414431 TI - Preoperative assessment of the acetabulum. AB - Preoperative assessment of the acetabulum requires a good knowledge of the normal acetabular anatomy, good plain radiographs, and a knowledge of the various defect patterns that may influence the surgeon's ability to achieve secure fixation of the component at surgery. In certain situations, special radiographic techniques are advantageous. PMID- 8414432 TI - Management of the deficient acetabulum using cementless fixation without bone grafting. AB - Based on the extensive experience with bulk femoral head augmentation of the acetabulum that began in 1973 and the disturbing incidence of failure that has been substantial and progressive, it is argued that sufficient justification, apart from exceptional circumstances, for the use of this reconstructive technique no longer exist. Instead, technical and technologic advances in the use and design of components that stabilize by biointegration now offer a more attractive alternative. PMID- 8414433 TI - Cemented fixation with bone grafts. AB - Loosening of the acetabular cup in cemented total hip arthroplasty is always accompanied by a loss of bone stock. Acetabular lesions can be reconstructed in several ways. Preoperative planning must be through, and specifically if graft procedures are considered, infection must be ruled out. The treatment of choice is a standardized cemented acetabular revision procedure with tight impaction of morsellized cancellous grafts. The clinical success of the technique is supported by the results of an animal experiment. PMID- 8414434 TI - Bone grafts in hip replacement surgery. The pelvic side. AB - Replacement arthroplasty of the hip may require restoration of bone stock to provide support for the acetabular implant and to restore anatomy and leg lengths. This article discusses the indications, surgical technique, results, and controversies of using bulk autograft bone to provide cup support for primary hip replacement in hip dysplasia. In addition, the use of allograft bone to restore bone stock in the revision situation is covered. The indications, surgical technique, and results of the use of both morsellized and bulk allograft bone are presented. PMID- 8414435 TI - Restoration of femoral bone stock in revision total hip arthroplasty. AB - The presence of femoral deficiencies in revision total hip arthroplasty may contribute to loosening, subsidence, and fracture. Between 1984 and 1990, 174 patients were treated with revision total hip arthroplasty, preformed by a standardized operative technique using cortical onlay strut allografts to restore the structural integrity of the femur and a calcar deficient proximally porous coated femoral prosthesis, composed of titanium alloy. The grafts united 98% of the time and showed evidence of revascularization and, in some cases, complete incorporation. Cortical onlay strut grafting in conjunction with a proximal load- bearing prosthesis is a viable method of reconstructing the structurally deficient femur in revision total hip arthroplasty. PMID- 8414436 TI - Revision arthroplasty of the proximal femur using allograft bone. AB - Revision arthroplasty of the hip may require restoration of bone stock on the femoral side to provide bone support for the new implant and restore anatomy and leg length. Large cortical defects can be reconstructed best with allograft bone because of the quality and quantity of the bone required. In this article, the surgical technique and results of cortical allograft as full circumferential and cortical strut grafts are presented. PMID- 8414437 TI - Contained morselized allograft in revision total hip arthroplasty. Surgical technique. AB - The current technique and instrumentation for contained morselized allograft with cement in dealing with femoral loosening associated with loss of bone stock is described. With a follow-up extending to 6 years, this method has proved effective in the management of these difficult problems. PMID- 8414438 TI - Custom-made segmental femoral replacement prosthesis in revision total hip arthroplasty. AB - Patients who present with failed total hip arthroplasty and significant proximal femoral bone loss pose a challenging reconstructive problem. When the integrity of the host proximal femur can be salvaged with cortical strut grafts, conventional long-stem femoral components can be used. If there is massive proximal femoral bone loss, hip salvage can be accomplished with a proximal femoral replacement prosthesis or an allograft-prosthesis composite. Although our early results with a proximal femoral replacement prosthesis were good, long-term results reveal a high rate of dislocation and acetabular component loosening, with an overall survivorship of 73% at 11 years. We have, therefore, limited the use of a proximal femoral replacement prosthesis to the elderly and inactive patient. We are encouraged by the preliminary results of total hip arthroplasty with a second-generation, modular, porous-coated, proximal femoral replacement prosthesis. Longer follow-up is required to determine whether these design modifications improve the results of this type of reconstructive procedure in revision total hip arthroplasty. PMID- 8414439 TI - Prevention of infection in the 1990s. AB - Deep sepsis after arthroplasty surgery represents a catastrophe. People remain the primary source of bacterial contamination. This paper discusses the potential sources of infection and reviews the appropriate measures required to reduce contamination and combat infection. PMID- 8414440 TI - Diagnosis of infection and the role of permanent excision arthroplasty. AB - The symptoms, signs, and investigation of the infected hip replacement are reviewed. Aspiration, combined with needle biopsy of the interface tissue, gives the best opportunity for establishing a bacteriologic diagnosis. The surgical technique for excision arthroplasty is described, and the complications of this procedure reviewed. The functional results of permanent excision arthroplasty are described in relation to the altered biomechanics of the hip pseudarthrosis. PMID- 8414441 TI - A temporary antibiotic-loaded joint replacement system for management of complex infections involving the hip. AB - Management of the infected hip replacement associated with substantial loss of bone, especially segmental loss of the proximal femur, is a challenging problem. In this report, the authors outline the development of an immediate-fit, custom made, antibiotic-selective replacement system for the hip and proximal femur, to help maintain limb length, joint stability, and mobility of the patient. Despite the initial relatively crude design, it was a valuable adjunct in the management of 15 consecutive cases. A more sophisticated and versatile system has since been developed for use in straightforward and complex cases. PMID- 8414442 TI - Exchange arthroplasty for infection. Perspectives from the United Kingdom. AB - A number of factors are considered with a bearing on outcome assessment and survivorship analysis. The results achieved in more than 440 exchange arthroplasties are discussed. Despite the apparent threefold increased risk of recurred infection associated with a one-stage approach, this statistic is an oversimplification of the many complex factors involved in the analysis of these results and in the decision between a one- and a two-stage approach to treatment, when presented with an infected hip replacement. The authors think that a one stage approach remains a reasonable alternative, given certain prerequisites. PMID- 8414443 TI - Unlicensed assistive personnel: a bane or boon for patient care? PMID- 8414444 TI - What are your risks, rights, responsibilities for bloodborne disease exposure in the workplace? PMID- 8414445 TI - RNs face risks as patient advocates. PMID- 8414446 TI - [Proteoglycans (their structure, function and role in liver diseases)]. AB - Proteoglycans are macromolecules containing a core protein to which glycosaminoglycan chains are covalently attached. The family contains several members with different structures and various functions. Some of them are elements of the extracellular matrix, while others are located to the cell surface playing important role in cell-cell and cell-extracellular matrix interactions. Present paper discusses the possible consequences of the alterations of proteoglycans observed in liver cirrhosis and liver tumors. It has to be emphasized however, that they are also involved in the pathomechanism of arteriosclerosis, Alzheimer-disease, immune diseases, arthritis, tumor progression and metastasis formation. PMID- 8414447 TI - [Preventive measures for early and late locomotor complications from thoracoplasty and myoplasty]. AB - Two basic methods of prevention of early- and late-occurring complications after thoracoplastic and myoplastic surgery are defined by the authors: I. The function protective surgical method and technique: by the retention of rib 1, the extramuscular periosteal thoracoplastic surgery, resection of as few ribs as possible, total performance of thoracoplastic and myoplastic surgery, epidural assuaging of pain. II. Perioperative physiotherapy and correct rehabilitative treatment, where gymnastics plays the most important role, and also combined application of the different special methods of physiotherapy, can also contribute to favourable results. 17 patients data have been processed and set out in the report. They show the efficiency of the applied methods. Planned and purposeful co-operation between the surgeon and the physiotherapist is needed in the medical treatment of patients. PMID- 8414448 TI - [Comparison of invasive and non-invasive measuring of blood pressure in patients following open heart surgery]. AB - Accuracy of blood pressure measurements with a new noninvasive ambulatory blood pressure monitor (Meditech KFT Budapest) was studied in 12 postoperative patients following open heart surgery. We compared 532 systolic and diastolic blood pressure recordings taken simultaneously both invasively and noninvasively. The values obtained noninvasively were subtracted from the corresponding values obtained invasively, and the differences were assessed according the British Hypertension Society recommendations. Values obtained with the two methods demonstrated good correlation (R: 0.85; p < 0.0001). 60 per cent of the differences in diastolic and 76 per cent of the differences in systolic blood pressure recordings fell into the range of +/- 5 mmHg. Thus the accuracy of diastolic blood pressure measurements was classified as "A" (characteristic fo the best equipment) and those of the systolic recordings was classified as "B" (characteristic of good equipment). Based on our findings the new blood pressure monitor provides accurate recordings and its use is highly recommended in the everyday practice. PMID- 8414449 TI - [Histological and clinical progression of a B-cell type non-Hodgkin lymphoma of low-grade malignancy with primary cutaneous manifestations]. AB - The development of primary cutaneous immunoblastic lymphoma, then its leukaemic phase was observed by authors in the course of disease of the young man with primary cutaneous centrocytic lymphoma. Authors think this case counts on interest because of the unusual appearance and course of centrocytic lymphoma. PMID- 8414450 TI - [Lymphangiomyoma of the thoracic duct]. AB - From the right superior mediastinum of a 31-year old woman the authors extirpated a lymphangiomyoma originating from the thoracic duct. The clinical feature and pathology of this rare lesions are discussed. This case is the first successfully removed thoracic duct lymphangiomyoma in Hungary. PMID- 8414451 TI - [Chemoluminescence induced by t-butyl hydroperoxide increases under the effect of cigarette smoke (preliminary report)]. AB - Cigarette-smoke in determined quantity was streamed through physiologic saline solution, or blood plasma or ex vivo excised rat lung. The solutions as well as the supernatants from lung tissue homogenate showed a significantly increased chemiluminescence after t-butyl hydroperoxide (BHP)-induction. The method reflects an expressive activity of free radical reactions caused by the smoke. PMID- 8414452 TI - [Ovarian cancer and the surgeon's knife. Early cancer of the ovary]. AB - The surgical aspect of ovarian cancer, based on the author's personal experience upon several hundreds patients and on the literature data, has been reviewed. Primary laparotomy is the cornerstone of the management of early stage ovarian cancer. This should include removal of the tumour and staging laparotomy. Better understanding of the tumour biological behavior is important to make a firm decision on conservative surgery in young patients with unilateral tumour who desire preservation of fertility. Rutin second-look laparotomy does not seem to be indicated in early stage disease. PMID- 8414453 TI - [Aeropollinologic and allergologic studies for the clarification of "poplar tree hay fever"]. AB - The authors performed allergen research on patients reported hay fever symptoms during the poplar pollen season (March), and in patients with hay fever symptoms during the time of the year that the seed hairs of the poplar trees are blowing in the air (May). Skin prick tests (Epipharm) and serum specific IgE tests (Epignost IgE Quick and Phadezym Populus deltoides RAST, Pharmacia) were performed on the basis of the pollen calendar of Szeged region on 30 patients. The pollen containt of the air was measured by means of a Lanzoni sampler. According to the pollen calendar of our region a large amount of grass pollens could be found in the air at the same time as the seed hairs of poplar trees are present (in May). The season of poplar pollen is in March in this area. Poplar pollen sensitivity was found on 8 patients. This is 6.8% of the total number of hay fever patients. They were found to be sensitive to other tree pollens too. The 23 patients complaining about hay fever symptoms in May, during the flaying of the cottons of the poplar trees in the air were all found to be sensitive to grass pollens. On the basis of our results the poplar pollen sensitivity is a relatively rare cause of hay fever. Our patients having complained about the seed hairs were all found to be sensitive to grass pollens. It seems that the grass pollens are the real cause of their disease. PMID- 8414454 TI - [Ursodeoxycholic acid therapy in biliary cirrhosis]. AB - Six female patients (mean age 48.5 years) suffering from primary biliary cirrhosis were evaluated during ursodeoxycholic acid (UDCA, Ursofalk, 10 mg/kg/day) therapy. The following examinations were performed: liver function tests, immunological studies [electrophoresis, immunoglobulin determination, T-B lymphocyte rate, antibody dependent cellularly cytotoxicity (ADCC), natural killer (NK) activity, blast transformation]. The patients showed a definite clinical improvement. Short term therapy (1 month) resulted in a decrease of gamma-GT, SGPT (ALT), SGOT (AST), bilirubin, alkaline phosphatase, cholesterol, triglyceride. The longer period of therapy (9 month) was associated with decrement in transaminase and gamma-GT levels. The statistically significant decrease of the IgG level, NK activity and spontaneous blast transformation during therapy suggest a potential immunosuppressive effect of the UDCA treatment. The authors believe that the use of UDCA could be effective in the treatment of primary biliary cirrhosis. PMID- 8414455 TI - [Use of current methods in the preservation of the health status of the population of the town of Szerencs (results of the Szerencs survey)]. AB - The author describes the methods and results of a health education project carried out by WHO and coordinated by the Outpatient Clinic and the adult and children health care district in Szerencs between 1989-1991. The results have indicated an extended presence of the cardiovascular diseases, malignant tumors, endocrine diseases and locomotor disorders in this town. The behaviour questionnaire have proved a lack of patterns of health promotion behaviour based on health knowledge in a significant part of the studied population. The results lead to the conclusion that health behaviour is also a result of socialization. PMID- 8414456 TI - [Bugat, the gardener]. PMID- 8414457 TI - [New aspects of the pathogenesis of progressive chronic polyarthritis (rheumatoid arthritis)]. AB - The author summarizes the newest results and knowledges which are related to the pathogenesis of polyarthritis chronica progressiva. Pursuing the composed target the feasible roles of gamma delta TcR T lymphocytes and stress proteins as well as that of CD5 positive B lymphocytes and IgG-glycosylation status are discussed. Reaching the end of the paper it is established that the solution of etiology and pathogenesis of polyarthritis chronica progressiva, in spite of the increasing theoretical knowledge, awaits for further extensive research work. PMID- 8414458 TI - [Monitoring respiratory mechanism and gas exchange in patients on respirators (side stream spirometry, capnography, oxigraphy, measurement of anesthetic agent concentration, pulse oximetry)]. AB - Author describes the working principles of the side stream spirometry, capnography, oxygraphy, anaesthetic agent concentration measurement, pulse oximetry, all of them incorporated in a new type of monitor, as well as the informations obtained by using them, concerning lung mechanics and gas exchange. The graphic and digital data, provided by the monitor are visualizing the correlations among the components of lung mechanics and are helping in setting up optimal ventilatory parameters, both during anaesthesia and during ventilation in an intensive care unit. The monitor approaches gas exchange from several aspects, which enhances the judgement of the efficacy of ventilation. PMID- 8414459 TI - [Retrospective survey of nursery-age children 5 years after Chernobyl]. AB - In their paper, the authors are dealing with the effect of the Chernobyl's nuclear catastrophe (1986) for the outcome of pregnancy and some parameters of the status of health. Their empirical study is based on a sample (sample size is 1168) of pregnant of the critical year 1986, and also based on a control sample (sample size is 1068) of the year 1985. They compared the five old year children's parameters of status of health between the critical and control groups. Their retrospective epidemiologic survey is extended to study some prenatal, perinatal and childhood parameters also. Finally, they summarise the biometric evaluation of their questionnaire survey. No significant difference of health conditions of the two groups defined above was found. PMID- 8414460 TI - [Smoldering myeloma]. AB - The authors present the history of a 74 year old woman. The patient was observed from 1972. Initially the bone marrow and serum protein dischanges were identical to multiple myeloma, therefore she was treated with melphalan-prednisolone regimen for one year. She was symptom free for 12 years with none of any treatment. Her bone marrow picture and serum protein aberrations were stabil. In 1985, the clinical signs and laboratory parameters of multiple myeloma arised. Major clinical symptoms, myelomatous bone lesions were detectable and the monoclonal protein level increased. In activation period repeated local irradiation and VCMP and VACP polychemotherapy was performed. Her history was typical to smoldering myeloma. The presentation of that disease is not available in Hungarian literature. The authors present the criteria of diagnosis, the therapeutic possibilities and the differential diagnosis of benign and malignant gammopathies. PMID- 8414461 TI - [Multiple endocrine neoplasia]. AB - The authors review present knowledge on multiple endocrine neoplasias types 1 and 2. They discuss in the light of recent literature data, the natural history, diagnosis, differential diagnosis and the optimal diagnostic approaches of these disorders. A short review on the therapeutic modalities draws attention to important differences between sporadic and inherited endocrine tumors. Finally, the authors emphasize recent developments about the genetic background and the importance of molecular biologic techniques, including family screening in the diagnosis of these disorders. PMID- 8414462 TI - [Surgical treatment of gallbladder diseases in patients over 70 years of age]. AB - The authors give a retrospective analysis of the results of 162 biliary operations performed in a five-year period on patients over 70 years of age. The mortality rate of 87 elective operations was 3.45%, of 75 acute operations 6.17%. During the same period 321 ERCPs and 120 ESTs were carried out in patients over 70, out of whom 2 were operated on and lost (mortality: 1.67%) because of bleeding after EST. The total mortality rate was 7.41% (n = 12). The circumstances of negative and positive choledochotomies were analysed together with the relationship between the operating time and the course of the disease. Out of the negative choledochotomised patients (n = 10) 3, out of the positive ones (n = 40) 5 patients died. For each patient over 70, to shorten the operative time and to prevent perioperative complications, the authors recommend the consideration of ERCP prior to biliary operation, and in certain cases, instead of elective biliary surgery, for bile duct stone, EST and stone extraction is advisable, leaving the calculous gallbladder "in situ". Further investigations are necessary, in the authors opinion, to decide whether prior to emergency operation on patients over 70 urgent ERCP and, when bile duct stone is diagnosed, EST with stone extraction is justified, too. PMID- 8414463 TI - [Autoimmune cytopenia in pernicious anemia]. AB - Authors report on their 5 patients with pernicious anaemia whose autoimmunocytopenia was diagnosed at different times after the diagnosis and treatment of their original disease. All of the patients were women. They have found autoimmune thrombocytopenic purpura in two cases, agranulocytosis in two cases and autoimmune haemolytic anaemia in one case. They compare their patients with the cases of literature. PMID- 8414464 TI - [Longitudinal socio-gerontologic geronto-epidemiologic study in Hajduszoboszlo (SZOLOVI, 1964-1991)]. AB - The tendencies of the demographic changes in Hungary, the trend and pace of the expected development of the age distribution and social stratification anticipated even in the early sixties that the problem of the ageing population (of the agriculture) would raise to national level and a considerable part of its consequent tasks will be the share of the national health care. So began in 1964 in Hajduszoboszlo a social gerontological-geroepidemiological study of those who were 60 and over and had agricultural profession. This study was developed into a longitudinal one (in the years 1986, 1990, 1991 and 1993), and is being continued even today. PMID- 8414465 TI - [Dr. Istvan Hamburger (1893-1952)]. PMID- 8414466 TI - ["Stroke"--"stabbing pain". Mythical elements in Hungarian disease terminology]. PMID- 8414467 TI - [Ovarian cancer and the surgical knife. Advanced ovarian cancer]. AB - Surgery of advanced ovarian carcinoma includes total abdominal hysterectomy, bilateral salpingo-oophorectomy, partial or total omental resection and removal as much secondaries as possible. Distinction between optimal and suboptimal debulking is important in terms of survival. This is based on the size of the diameter of the largest residual tumour following primary surgery (< 2 cm vs > 2 cm). The value of lymphadenectomy is still controversial. However, it should be explored in prospective studies. The role of second-look laparotomy in advanced ovarian carcinoma remains debatable. Secondary debulking appears to be beneficial only if it is followed by adjunctive chemotherapy. PMID- 8414468 TI - [Iatrogenic sinus node disease and iatrogenic AV block]. AB - The appearance of long lasting bradycardia due to damage of the sinus node (iatrogen sick sinus syndrome; or the atrioventricular conduction system, iatrogen AV block) is not a rare event after open heart surgery. In the course of 5093 heart operations the development of iatrogen sick sinus syndrome was observed in 234 patients (4.6%) and iatrogen AV block in 91 cases (1.8%). The incidence rate of iatrogen sick sinus syndrome or iatrogen AV block was quite divergent depending mostly on the type of operation. Comparing the data of our earlier (1977-1982) and later (1983-1991) cardiac surgical interventions, the prevalence of iatrogen sinus node disease and iatrogen AV block seems to decrease, mainly due to the progress in techniques of cardiac surgery. With the appearance of iatrogen sick sinus syndrome or AV block, urgent temporary pacing is indicated to prevent the deleterious hemodynamic effect of bradycardia. In the cases of persistent iatrogen sick sinus syndrome and iatrogen AV block, permanent pacemaker implantation is needed. It is remarkable that although the incidence rate of iatrogen sick sinus syndrome is greater than that of iatrogen AV block, in iatrogen sick sinus syndrome the regression is quite frequent, while in iatrogen AV block the 2nd or 3rd degree AV block is usually permanent. We performed pacemaker implantation in 56 cases of iatrogen sick sinus syndrome (24%) and in 57 patients of iatrogen AV block (63%). With pacemaker therapy the outcome of iatrogen sick sinus syndrome and iatrogen AV block is very favourable. PMID- 8414469 TI - [Myofibrosis in childhood. Differential diagnosis in chest pain]. AB - It is emphasised in connection with the differential diagnosis of chest complaints that after excluded the serious and urgent conditions, a searching for painful muscle bundles and trigger points at the characteristic locations in the muscles can explain the origin of the complaints. By finding of myofibrosis, other expensive or invasive diagnostic tests can be omitted and the patient gets rid off his or her claims. PMID- 8414470 TI - [Effect of paracetamol on the acid hemolysis of erythrocytes, with special reference to the intracellular glutathione level]. AB - The authors give account of their study concerning the effect of paracetamol on the acid haemolysis, and glutathione content of human erythrocytes. In vitro, paracetamol decreased intracellular glutathione content in a dose dependent manner. This may lead to an increased sensitivity of erythrocytes to the haemolytic effect of hydrochloric acid, what may explain the dose dependent decrease of time of haemolysis, presented in form of dynamic haemolysis curve. In vivo four hours following the oral intake of 500 mg paracetamol, although the intracellular glutathione content did not decrease significantly, shortening of the time of acid haemolysis could be demonstrated in more than half of the persons studied. No haemolysis was caused by the given dose of the drug. PMID- 8414471 TI - [Effect of penicillamine D, nitric oxide, or both?]. PMID- 8414472 TI - [Information about the "scarcity" of disulfiram implant]. PMID- 8414473 TI - [Irene Zach, the new president of the Austrian Nursing Association]. PMID- 8414474 TI - [Austrian Nursing Association-provincial sector Vienna/Burgenland: preparations for the congress, a two year project. Interview by Harald Verworner]. PMID- 8414475 TI - [Lumbar spinal stenosis. Classification and clinical presentation]. AB - A review is presented of the nomenclature, taxonomy and clinical presentation of the lumbar canal and lateral stenosis. A historical review of the literature illustrates the growing importance of this clinical entity in the narrow spinal canal. The definition of Verbiest is introduced and explained. It distinguishes two main groups of spinal stenosis: congenital and developmental stenosis. Developmental stenosis contains several subgroups. Special attention is paid to "central" lumbar canal stenosis and lateral spinal stenosis. Both entities are discussed with a description of the etiology, anatomy, and symptomatology. The importance of nerve root infiltration as a diagnostic tool in asymptomatic radiculopathy is emphasized. PMID- 8414476 TI - [Clinical aspects of spinal stenosis]. AB - Spinal stenosis is a common problem in the elderly population. Thanks to better knowledge it is being diagnosed more frequently and in many cases successfully treated by surgery. Clinically, patients with a lumbogluteal and in later stages bilateral sciatic pain syndrome can be differentiated from patients with monoradicular pain. Typically, the posture deteriorates with walking distance. Spinal stenosis must be differentiated from peripheral vascular disorders and compressive disc herniation. The possibilities of conservative therapy are limited and their effect is lasting only in mild cases. PMID- 8414477 TI - [Radiological assessment of degenerative lumbar stenosis]. AB - The evaluation of central and lateral lumbar spinal stenosis is equally well performed by CT, CT-myelography and MRI. Stenosis can be quantified and localized in order to optimize orthopedic treatment. Each of these modalities has inherent strengths and weaknesses. CT-myelography is still the best modality for evaluation of central spinal stenosis because absolute stenosis can be more precisely quantified than by plain CT and MRI. Myelography can image the entire spine and localize stenosis that is induced by functional positions. Lateral lumbar spinal stenosis can be localized at the lateral recessus or more distally at the neural foramina. Stenosis of the lateral recessus occurs only at the lower levels of the lumbar spine. Rarely stenosis at the entry of the lateral recessus can be missed by MRI and is only depicted by thin section (1-2 mm) CT. The anatomy and therefore stenosis of the neural foramina are exquisitely demonstrated by direct sagittal MRI. PMID- 8414478 TI - [Histological changes in the ligamentum flavum in patients with spinal stenosis]. AB - In 21 patients with spinal stenosis the ligamentum flavum was removed, histologically processed and the calcification was measured with an image analyzing computer. Nearly all the ligaments were calcified: on average 0.17%, with a maximum of 3.8%. There was a significant correlation between age and calcification. We compared these findings with those of a group of "normals". There we only found minimal calcification in 3 of 20 segments (0.025%, 0.049%, 0.22%). As these people were on average 85 years of age, this can be looked upon as normal degenerative changes. This illustrates the important role of calcification of the ligamentum flavum for spinal stenosis. PMID- 8414479 TI - [Anamnesis and myelography in the preoperative assessment of lumbar spinal stenosis. Results of a postoperative follow-up study]. AB - Between 1987 and 1991, 33 patients with spinal stenosis of the lumbar spine were treated by decompression (33 patients) and posterior fusion (30 patients). Indication for decompression was based on case history and lumbar myelography with flexion/extension views. At follow-up 1-5.5 years later, 28 patients were happy with the results of the treatment and would be willing to be operated on again in a similar situation. Two other patients also presented objectively good results, but were dissatisfied for reasons not related to the operation. Our study shows that myelography and case history are adequate investigations for determination of the level of pathology and for making a decision about operative decompression in spinal stenosis of the lumbar spine. CT or MRI are only needed if the symptoms of the patient are not explained by the myelogram. Although MRI is advocated as the investigation of first choice for lumbar spinal stenosis, we still prefer the myelography, which is easier to interpret during the operation. Our study also shows that operative treatment of spinal stenosis is very rewarding, since 9 out of 10 patients will have good results. We usually combine decompression and fusion. Decompression alone is only performed in patients without any back pain and with stable motion segments after adequate decompression. PMID- 8414480 TI - [Degenerative lumbar scoliosis and spinal stenosis]. AB - Of the types of degenerative lumbar spine deformities, the combination of scoliosis and spinal stenosis is one of the most disabling and progressive conditions in elderly patients. Therapeutic resignation is frequent. In spite of old age and severe osteoporosis 31 patients were treated operatively by reduction, fusion and decompression. There were 24 good results, 2 fair and one poor result. During the postoperative 30-month follow-up period (1-5 years), 4 patients died from unrelated causes. There were no serious primary complications; 3 patients had a reoperation. All patients were living independently at their homes and had a walking capacity of more than 2 km. The good overall result is attributed to the selection of patients, which is the most important factor for the outcome. Nine criteria for patient selection are listed. In multisegmental lumbar spine fusions in elderly patients, the critical areas are sacral fixation of the implant, a loss of lumbar lordosis and, if the fusion is performed from L2 caudad, vertebrae L1 and T12 being located between the lumbar spondylodesis and the stiff thoracic spine. PMID- 8414481 TI - [Significance of simultaneous fusion and surgical decompression in lumbar spinal stenosis]. AB - In a prospective study 45 patients with clinically relevant spinal stenosis of the lumbar spine were randomized in three groups. Group 1 underwent isolated decompression without fusion, group 2 had decompression and selected fusion, and group 3 underwent fusion of all decompressed segments. Patients with previous surgery of the lumbar spine or obvious instability, such as spondylolisthesis, were excluded from the study. The results were dominated by the relevant clinical improvement in the symptoms of spinal stenosis in all three groups. There were no statistical significant differences in the clinical results between the patients with and without fusion. We conclude that in the absence of obvious segmental instability, no fusion is required for decompressive surgery in degenerative lumbar spinal stenosis. PMID- 8414482 TI - [Lumbar spinal stenosis in systemic diseases and in hereditary syndromes]. AB - In the general category of congenital spinal stenosis, there is a group of patients with particular features: patients suffering from a hereditary or systemic disease with congenital narrowing of the spinal canal. Evaluation of their neurological symptoms is influenced by the peculiarities of the underlying disease. The tendency toward paralysis and paraplegia is much higher in those patients. Attention must especially be given to technical details when operating on achondroplastic dwarfs. Early, most extensive surgery is recommended. We report our experience between 1987 and 1992 with operations on 6 cases of stenosis of the lumbar canal with hereditary or systemic disease. PMID- 8414483 TI - [Lumbar spinal stenosis. An overview 50 years following initial description]. AB - About 50 years after the first descriptions of lumbar stenosis and its most frequent symptom, neurogenic intermittent claudication, this update gives an overview of present-day concepts of the disease and of new experience in this area. Stenosis of the central and lateral lumbar spine is chiefly held to be one of the results of segmental degenerative instability throughout the several stages of spondylosis. The degenerative process of the spine has its starting point in regressive changes of the disc. This leads to instability of the motion segment, which explains the pathophysiological dynamics of the stenosis and its symptoms, including intermittent neurogenic claudication. Segmental instability is the crucial lesion causing all the changes in the degenerative process, which are not to be considered as separate entities but as part of the dynamics of the same disease. The spine's congenital anatomic individual patterns, which confirm the population thinking of the evolutionary biology (since they are different from one motion segment to the other) help to determine the outline of single cases. The disparity between radiological and clinical patterns is pointed out: severe stenosis may be asymptomatic or cause just modest monoradicular trouble as well as serious multiradicular deficit. The reason for the discrepancy is unknown. One must be careful to avoid surgery on a silent, purely radiological stenosis. Experience of more than 15 years confirms the uselessness of performing a complete laminectomy to achieve sufficient decompression. The author's method of selective decompression is described briefly. The English term "undercutting decompression" runs the risk of being misunderstood, since it is also used for rather destroying procedures. Finally, we point out that surgery for spinal stenosis must resolve both root compression and degenerative instability in the majority of cases. In most cases of lumbar stenosis, if decompression alone is performed, only the consequence, and not the cause of the disease, segmental degenerative instability, is treated. Osteophyte formation is an attempt by nature to stabilize the motion segments by stiffening its components. Spinal fusion tries to achieve the same effect. For most cases of spinal stenosis, we suggest our own technique, which combines safe and preserving ("selective") decompression according to Benini [1,7] with the translaminar screw fixation of Magerl [7]. In cases of degenerative spondylolisthesis, however, transpeduncular fusion is mandatory. PMID- 8414484 TI - [The effect of ultrasonography screening of hips in newborn infants on femur head necrosis and the rate of surgical interventions]. AB - We compared two similarly sized groups of sonographically unstable or decentered hips with comparable initial findings, both treated by standardized conservative primary treatment. The results show that an early definite diagnosis improves the prognosis. If built on a secure initial diagnosis rationally grounded and efficient conservative therapy can be initiated and followed through on. Since the treatment is started extremely early, the hip can benefit from the considerably greater potential for spontaneous development present in the first three months of life. In this way we can shorten the treatment time considerably and also complete it at a younger age. Finally, early treatment also reduces the risk of the complication of necrosis of the femoral head and significantly decreases the rate of surgical procedures. Only a sufficiently well executed and comprehensively organized sonographic neonatal screening program can produce a definite very early diagnosis and with it almost 100% successful early treatment of all grades of disturbances of hip maturity. Sonographic screening of the newborn is a very important piece in the jigsaw of prophylaxis and primary prevention in our public health care program. PMID- 8414485 TI - [Results of ultrasonographic screening. An epidemiological study]. AB - The possible influence of early harness treatment for hip dysplasia--a treatment made possible by hip sonography according to Graf--on the incidence of hip luxations was examined in an epidemiological study. In a region with 8,000 births per year three periods were investigated: the pre-ultrasonographic era with 19 luxations per year (2.2/1000), the introductory era with 11 (1.3/1000) and the last period with 6 luxations so far (0.8/1000). A diagnosis can now be made at an earlier time due to the possibility of viewing the hip with sonography, a fact which significantly reduces the amount of hip luxations (pre-ultrasonographic era versus screening era: P = 0.00002), verified by the significant decrease in open and closed reductions. PMID- 8414486 TI - [The differential diagnosis of juvenile hip pain in the ultrasonographic picture. Transient coxitis. Legg-Calve-Perthes disease, epiphysiolysis of the femur head]. AB - The efficiency of ultrasonography was tested in examinations of 117 children with hip pain, transient synovitis (n = 41), Legg-Calve-Perthes disease (n = 61) and epiphysiolysis capitis femoris (n = 15). The sonographic criteria for the evaluation are presented. In the case of transient synovitis the effusion persisted for < 2 weeks. In the early stage of Legg-Calve-Perthes disease all patients had had capsular distension for > 6 weeks. A differential diagnosis between transient synovitis and Legg-Calve-Perthes disease is possible. In the case of epiphysiolysis capitis femoris, it is possible to diagnose slipping of the epiphysis and capsular distension. The significance of synovitis in the pathogenesis of Legg-Calve-Perthes disease is discussed. PMID- 8414487 TI - [Ultrasonographic appearance of the rotator cuff in elderly subjects]. AB - Some special points must be taken into consideration when diagnosing complaints of the shoulder joint in older patients by means of sonography. Such patients not only have a higher incidence of systemic disease, but also--as we know from autopsies--a large percentage of rotator cuff tears. These changes--with the exception of small partial tears--can be recognized sonographically with a high degree of certainty. Prospective sonographic studies have provided us with information that the incidence of rotator cuff tears rises from 25% in patients in their fifties to 50% in patients in their seventies. These studies enabled us to explain the hitherto unknown clinical relevance of these changes. In general, the patients complained of little pain, but when large tears were present, there was usually a clear loss of function, which however was astonishingly well tolerated. These findings would call for caution in indicating complicated reconstructive surgery. PMID- 8414488 TI - [Sonographic diagnosis in shoulder joint instability. Possibilities and limitations]. AB - Ultrasound evaluation expands the possibilities of diagnostic imaging techniques for patients with shoulder instabilities. Static ultrasound investigation documents the acute or chronic dislocated shoulder. Dynamic evaluation demonstrates the direction of instability. Concomitant Hill-Sachs lesions or rotator cuff tears can be diagnosed with high diagnostic accuracy. Clinically relevant documentation of labrum defects (Bankart lesions) by ultrasound is not possible with the technique currently available. PMID- 8414489 TI - [Ultrasonographic diagnosis in inflammatory-rheumatic changes of the shoulder joint]. AB - Inflammatory rheumatoid changes of the shoulder joint are often diagnosed rather late, so therapy is largely directed towards pain reduction and less towards maintenance of full joint function. When considering the mutilating type of chronic polyarthritis, it becomes evident how important the role of the shoulder joint is in the patients performance of basic everyday activities. In this paper the important structures of the shoulder joint that can be visualized by ultrasound are described as to their rheumatological significance, and pathological findings are discussed along with the appropriate therapy. PMID- 8414490 TI - [Current developments in ultrasonography of the meniscus]. AB - Sonography of the meniscus is a new diagnostic method, the instructive value and clinical relevance of which are subject to controversy. Sonographic assessment of the meniscus requires the use of focusing that can cope with the different anatomical conditions of the individual parts of the meniscus and can largely eliminate artifacts by means of wide sound-wave propagation. The dynamic examination technique allows the identification of both longitudinal and flap tears, which are relatively easy to diagnose, and of tear formations that are harder to visualize, such as horizontal and transversal tears. A sonographic criterion of a tear is the detection of a clearly visible, strong-signal double or single-stroke reflection pattern that can be traced in several section planes. While sonographic examination allows location of the tear, definite conclusions as to the shape of the tear are not possible on the basis of the echo pattern. In the literature, the statistical results, depending on the patient group and technical equipment, vary between 76% and 100% for sensitivity and between 50% and 97% for specificity, 74% and 93% for accuracy and between 61% and 95% for the positive prediction value. Sonographic examination of the meniscus is an easily available, noninvasive imaging technique that, as a supplement to clinical findings, can be used to optimize preoperative diagnosis and to check the indication for arthroscopy. Careful consideration of the technical requirements and systematic performance of the dynamic examination should lead to further improvement in the examination results and to growing clinical significance in the future. PMID- 8414491 TI - [Ultrasonographic monitoring of conservative-functional treatment of rupture of the Achilles tendon]. AB - Rupture of the Achilles tendon has become more common in recent years. Operative repair is the treatment generally used but non-surgical therapy is being accepted by an increasing number of surgeons because of the reports of good results with conservative treatment. Ultrasonographic examination not only allows an exact diagnosis to be made in muscle and tendon injuries, but also observation of the healing processes. Therefore, conservative functional treatment of rupture of the Achilles tendon can be used with more success, which is a very comfortable concept for these patients. We present a series of 22 subcutaneous ruptures of the Achilles tendon treated non-operatively to emphasize the value of conservative functional therapy with ultrasonographic monitoring. After immobilization of the lower leg by a walking cast in the equinus position for 3-6 weeks, the functional treatment follows using a decreasing heel lift and physiotherapy. Ultrasound examinations were performed frequently to recognize new diastasis early. We observed complete healing in all patients, absence of complications and excellent functional results. The morbidity of the patients was lower than with operative treatment and no hospital stay was needed. In conclusion, in our opinion ultrasound-controlled conservative functional treatment of rupture of the Achilles tendon should be used more frequently in future. PMID- 8414492 TI - [Ultrasonographic anatomy and ultrasonographic assessment of the transverse arch of the foot]. AB - A new and original sonographic technique to document the morphology of normal feet and splay feet is presented: a standard sectional plane cuts the transverse arch from the plantar side using the sesamoids of the first metatarsal bone and the head of the fifth metatarsal bone as reference points. A 5-MHz linear transducer and a rather simple custom-made platform with an integrated gel-pad are required in order to get reproducible results. The morphology of the transverse arch can be characterized by the "transverse arch index Q" as the parameter of the relative height of the transverse arch. Based on this transverse arch index Q, splay feet can be distinguished from normal feet in a statistically significant way (p < 0.01). Furthermore, grading of splay feet is possible and a documentation of rigidity/flexibility of the forefoot in splay feet; thus, the decision on whether to use conservative or operative treatment can additionally be based on sonographic documentation, and the results of surgical reconstruction of the transverse arch can be checked and documented sonographically. PMID- 8414493 TI - Two novel functions associated with the Rel oncoproteins: DNA replication and cell-specific transcriptional activation. AB - The v-Rel oncoprotein and its cellular homolog c-Rel belong to the Rel/kappa B family of transcription factors. Members of this family share extensive sequence similarity in their N-terminal halves, a region referred to as the Rel Homology Region (RHR), bind to NF-kappa B DNA motifs and form heterodimers with one another. Whereas c-Rel activates transcription of kappa B-linked genes, v-Rel behaves as a dominant-interfering mutant of c-Rel- and kappa B-mediated transcription activation. Here we describe two novel activities of the Rel oncoproteins. One induces kappa B-site dependent stimulation of polyomavirus (Py) DNA replication and maps to the N-terminus of the RHR, a region where no transcription activation function was detected. This activity is common to v-Rel, c-Rel, p52 (p49/lyt10), RelA (p65) and the p50 subunit of NF-kappa B. The second promotes transcriptional activation in undifferentiated F9 cells and maps 3' to the RHR, a region essential for the transforming activity of v-Rel. PMID- 8414494 TI - Retroviral infection can abrogate the factor-dependency of hematopoietic cells by autocrine and non-autocrine mechanisms depending on the presence of a functional viral oncogene. AB - The mechanisms responsible for abrogation of the growth factor-dependency of a hematopoietic cell line were investigated. FDC-P1 cells were infected with retroviral constructs containing the neo gene and either a wild-type or a temperature-sensitive v-src oncogene. v-srcwt abrogated the factor-dependency of these cells since each G418r colony gave rise to factor-independent cells and no autocrine growth factor activity was detected. Moreover, the vast majority (< 99%) of cells infected with the v-srcts mutant gave rise to conditional factor independent cells. Therefore a functional v-src gene product was required for growth factor-independence which occurred by a non-autocrine mechanism. A minority of factor-independent cells which arose after v-srcts infection, grew at the non-permissive temperature and one-half secreted granulocyte/macrophage colony stimulating factor (GM-CSF) which supports the growth of the parental cells. Since the v-srcts viral stock contained a helper virus, Murine Leukemia Virus (MuLV), the ability of this virus to relieve factor-dependency was examined. A low frequency of factor-independent transformants was recovered after MuLV infection and one-half secreted GM-CSF. Therefore, retroviruses such as MuLV which lack an oncogene, can transform cells by stimulating autocrine growth factor secretion. Subsequent experiments performed with helper-free v-src preparations indicated that they could abrogate factor-dependency directly by a non-autocrine mechanism. These results demonstrate that a hematopoietic cell line can be transformed by two different mechanisms after retroviral infection and may be relevant for understanding hematopoietic cell transformation after persistent viral infection. PMID- 8414495 TI - Characterization of the ret proto-oncogene products expressed in mouse L cells. AB - The ret proto-oncogene (proto-ret) encodes a receptor type tyrosine kinase with a cadherin-related sequence in the extracellular domain. To investigate whether the proto-Ret protein functions as a cell adhesion molecule like cadherins, we transfected the human proto-ret gene fused to the SV40 promoter or cytomegalovirus (CMV) promoter into mouse L cells in which cadherins are not expressed. Three transfectants with high levels of expression of the proto-Ret proteins were obtained. The proto-Ret proteins were expressed as 150 kDa and 170 kDa glycoproteins in transfectants as observed in human neuroblastoma cells. Cell fractionation experiments revealed that the 170 kDa protein but not the 150 kDa protein was detected predominantly in the plasma membrane fraction, indicating that the 170 kDa protein represents the mature glycosylated form of the proto-Ret protein present on the cell surface. Both 150 kDa and 170 kDa proto-Ret proteins showed tyrosine kinase activity in immunocomplex kinase assay. It is known that cadherins have Ca(2+)-dependent homophilic binding activity and are resistant to trypsinization in the presence of Ca2+. When L cells expressing the proto-Ret proteins were treated with trypsin in the presence of Ca2+, the 170 kDa protein was resistant to its digestion. On the other hand, it was completely digested in the presence of EGTA, suggesting the possibility that the proto-Ret protein interacts with Ca2+ like cadherins. However, the transfectants did not show clear adhesive properties in cell aggregation assays. PMID- 8414496 TI - A complete description of the EGF-receptor exon structure: implication in oncogenic activation and domain evolution. AB - In this paper, we report that the chicken Epidermal Growth Factor Receptor (EGF R), encoded by the proto-oncogene c-erbB, is comprised of 28 exons and spans over 75 Kb. The four previously identified domains which make up the extracellular ligand binding region of the receptor are coded for by two copies of a 300 amino acid repeat. We have demonstrated that the 3' end of each repeat coincides with the 3' end of the last exon making up the repeat. This alignment suggests that an exon-duplication may have occurred in the ligand-binding region of the gene. The transmembrane domain is encoded within a single exon and the exon boundaries of the catalytic domain closely match those defined by structural homology with other kinases. Along with the 54 chicken splice sites, the region 5' to the first exon was also sequenced. The proposed promoter region is greater than 70% GC, contains five repeats of the consensus Sp1 binding sequence and does not have a CCATT or TATA box. In addition to the presence of these characteristic housekeeping features, expression analysis confirms the promoter activity of this region and four sets of TCC repeats similar to those found in the human EGF-R promoter have been identified. To our knowledge this represents the first complete description of the exon-intron structure of the EGF-receptor family. The elucidation of the EGF-R exon structure provides insight into the domain evolution of the receptor-kinases and the mechanisms for the oncogenic conversion of EGF-R in erythroleukemias and glioblastomas. PMID- 8414497 TI - HTLV-I encoded Tax in association with NF-kappa B precursor p105 enhances nuclear localization of NF-kappa B p50 and p65 in transfected cells. AB - NF-kappa B is a heterodimeric protein composed of two subunits, p50 and p65, and is sequestered in the cytoplasm as an inactive form through the association with an inhibitory protein, I kappa B. In the present study, the effect of HTLV-I encoded Tax on the exogenously expressed NF-kappa B p105, which encodes a precursor of p50, was investigated using a COS-7 expression system. When p105 was expressed in COS-7 cells, the precursor p105 and the processed p50 were retained in the cytosolic fraction by associating each other, and p50 was not detected in the nuclear fraction. In the cells co-expressing p105 and Tax, the p50/p105 ratio in the cytosolic fraction reduced with an induction of p50 in the nuclear fraction, which gave rise to a significant increase in NF-kappa B binding activity. Enhancement of NF-kappa B binding activity was not observed by adenovirus encoded E1A and bovine papilloma virus encoded E2, and correlated well with the ability of Tax to associate with p105. When p105 delta X which lacks the repeats of the ankyrin motif was expressed in this system, p105 delta X and processed p50 were detected in the nuclear fraction and p50/p105 delta X ratio was not affected by co-expression with Tax. In the same expression system, exogenously expressed NF-kappa B p65 was retained in the cytoplasm by p105 and further expression of Tax allowed entry of p65 in the nucleus. These results suggest that, in this model system, p105 acts as an I kappa B to sequester p50 and p65 in the cytoplasm and that Tax by inhibiting I kappa B activity of p105, enhances nuclear localization of p50 and p65. These findings raise a possibility for a novel mechanism for the induction of NF-kappa B in the nucleus by Tax. PMID- 8414498 TI - Phosphorylcholine: a novel second messenger essential for mitogenic activity of growth factors. AB - Growth factor stimulation of quiescent cells induces a series of intracellular early and late events that ultimately lead to DNA synthesis and cell division. We describe here that production of phosphorylcholine is an essential component of the late events involved in the induction of DNA synthesis by platelet-derived growth factor (PDGF-BB), a prototype mitogen for fibroblasts. Moreover, phosphorylcholine itself is mitogenic when added exogenously to NIH3T3 cells, further indicating its role as a crucial intracellular messenger for DNA synthesis. Choline kinase, the first step in the route of phosphatidylcholine synthesis appears to be the critical regulatory enzyme in phosphorylcholine production, indicating that regulation of choline kinase represents a key step during mitogenic stimulation. We also describe that several growth factors (PDGF AA, basic FGF, EGF and phorbol esters) rely on their ability to generate phosphorylcholine for their proliferating activity. In contrast, DNA synthesis induced by serum did not require phosphorylcholine. Moreover, the requirement for phosphorylcholine production in PDGF-stimulated cells can be over-ruled by addition of insulin. Thus, cell proliferation in NIH3T3 cells can be triggered off by alternative pathways and one of them involves generation of phosphorylcholine. PMID- 8414499 TI - MHC class I expression in HPV 16 positive cervical carcinomas is post transcriptionally controlled and independent from c-myc overexpression. AB - Squamous cell carcinomas of the uterine cervix (n = 23) were selected for the presence of human papillomavirus type 16 (HPV 16) using the polymerase chain reaction (PCR). Localization of transcripts coding for the E7 protein was demonstrated in neoplastic cells with RNA in situ hybridization. Consecutive tissue sections were investigated for expression of the major histocompatibility complex class I (MHC-I) and c-myc using immunohistochemical double staining procedures, since a role has been suggested for the c-myc protein in MHC-I down regulation and c-myc overexpression has been described in cervical carcinomas. Reduced expression of class I heavy chains was observed in neoplastic cells from 18 out of 23 carcinomas (78%). Varying levels of c-myc overexpression were observed in 12 carcinomas (52%), from which four showed positive MHC-I expression in c-myc overexpressing cells. In the remaining eight c-myc overexpressing carcinomas MHC-I down-regulation was observed. Additional RNA in situ hybridization with class I heavy chain locus-specific RNA-probes revealed presence of class I mRNAs in those neoplastic cells that show negative staining for MHC-I protein. These data strongly indicate that MHC-I down-regulation in cervical carcinomas involves post-transcriptional mechanisms, not directly related to E7 transcription and overexpression of c-myc. PMID- 8414500 TI - Rb may act as a transcriptional co-activator in undifferentiated F9 cells. AB - The reversible interaction of the retinoblastoma susceptibility gene product (Rb) with the cellular transcription factor E2F has recently been demonstrated. Activation of the adenovirus E2a promoter by the products of the viral E1a gene correlates with the ability of both early E1a proteins to sequester Rb, thereby releasing E2F from inactive complexes with this protein. The E2a promoter is also efficiently stimulated by a product (17.5 kDa) of the viral E4 gene. The specific interaction of this E4 protein with E2F results in the formation of complexes that bind cooperatively to the two neighboring E2F binding sites in the E2a promoter. We have previously shown that in undifferentiated F9 cells (F9EC) the E2a promoter is refractory to E2F-mediated activation by E1a, but not by E4. Using both band-shift and transfection experiments, we now demonstrate (i) that in F9EC cells the E4 product, in combination with E2F, recruits Rb into a stable multiprotein complex and (ii) that in these undifferentiated cells, as opposed to their differentiated counterpart, Rb is actively involved in the transcriptional stimulation of the E2a promoter by E4. Our results suggest that, depending on the cell state, Rb may behave either as a transcriptional activator (F9EC cells) or as a transcriptional inhibitor (differentiated F9 cells). PMID- 8414501 TI - Modulation of cell growth, p34cdc2 and cyclin A levels by SV-40 large T antigen. AB - Immortalization of rat lung epithelial cells by either wild-type SV-40 T antigen, a mutant form of T antigen that cannot bind pRb, or a temperature-sensitive T antigen increased by five- to 20-fold the steady state levels of p34cdc2 and cyclin A, positive regulators of progression through the cell cycle. Increased abundance of p34cdc2 was not accompanied by equivalent increases in cdc2 mRNA, indicating that increased expression of p34cdc2 is due, at least partially, to post-transcriptional mechanisms. Levels of p34cdc2 and cyclin A protein in cells immortalized with a temperature-sensitive T antigen remained elevated at the restrictive temperature unless T antigen was reduced to levels significantly below those where proliferation ceased, indicating that these two functions can be dissociated. These results show that SV-40 T antigen can dramatically enhance the expression of certain cell cycle regulatory proteins by mechanisms that are independent of pRb binding and cell growth status. PMID- 8414502 TI - Analysis of p53 transactivation through high-affinity binding sites. AB - Alterations or elimination of the p53 protein is frequently occurring during human carcinogenesis. Overexpression of wild-type p53 has a profound growth inhibitory effect on many cell lines, including strong and apparently non sequence specific repression of a number of promoters. Consistent with the hypothesis that it acts as transcriptional regulator, wild-type p53 protein binds DNA and activates transcription of several promoters. We have studied DNA binding and transactivation (TA) properties of human wild-type and mutant p53 proteins representing four major mutational hotspots. DNA-gel retardation was used to detect specific p53-DNA complexes in nuclear extracts, with radiolabelled oligonucleotides representing high affinity p53-binding sites (HBS) as a probe. p53-specific complexes were identified by competition with unlabelled 'self' oligos and by double band-shifts in the presence of anti-p53 antibodies. To show transactivation by p53, TK promoter-driven CAT reporter gene was placed 3' of the p53-binding site. CAT activity was assayed after co-transfection of reporters with either wild-type (WT) or mutant p53 expression constructs into human cells that do not express p53 (SKOV3). We found that wild-type p53 has strong transactivating effect on the reporter. All mutants, with the exception of His273, were inactive in TA-assay. p53 is a target of several oncogenes found in DNA tumor viruses. We examined the effect of either SV40 T-ag or 55 kDa EIB protein of Ad5 on DNA binding and transactivation by p53 in transformed COS-1 and 293 cell lines, respectively. COS-1 extracts produced strong p53-dependent band shift of the HBS oligos, that was doubleshifted by anti-p53 but not anti-T-ag antibodies, indicating that T-ag is not part of the complex. COS-1 cells had a high level of WT p53-dependent expression of transfected CAT reporter, indicating the presence of transactivation-competent p53, acting through the HBS element. In human Ad-transformed 293 cells, endogenous p53 was also transactivation competent and capable of DNA binding. In summary, we found efficient transactivation of HBS motif by WT and His273-p53. Studies of COS-1 and 293 cells suggest that a proportion of p53 in transformed cells display wild-type DNA binding and TA properties and that expression of transcriptionally inactive mutant p53 proteins in these cells does not interfere with WT-dependent transactivation. PMID- 8414503 TI - Stabilization of wild-type p53 in human T-lymphocytes transformed by HTLV-I. AB - Adult T-cell leukemia/lymphoma is an aggressive malignancy associated with infection by the human T-lymphotropic virus type-I (HTLV-I). We now demonstrate that p53 expression is elevated in the HTLV-I-transformed T-lymphocyte lines C81, MT-2, MT-4 and HUT 102. In pulse-chase experiments, the p53 protein demonstrated a prolonged half-life of 2 to 8 h in HTLV-I-transformed cells compared with 0.5 to 1.0 h for wild-type p53 in primary human and murine fibroblasts, or human peripheral blood lymphocytes. In cell lines C81 and HUT 102, which exhibited the longest p53 protein half-life, the wild-type-related PAb1620 epitope was detected at reduced levels. The PAb240 mutant-related p53 epitope was not detected in any of the transformed cell lines. By direct sequence analysis of RT-PCR products, the entire p53 cDNA coding sequence was determined to be wild-type in all four cell lines. Stabilization of wild-type p53 may represent its functional inactivation and contribute to lymphocyte transformation by HTLV-I. PMID- 8414504 TI - Expression of the TAL1 proto-oncogene in cultured endothelial cells and blood vessels of the spleen. AB - The TAL1 proto-oncogene encodes a basic helix-loop-helix (bHLH) protein that has been implicated in the pathogenesis of T-cell acute lymphoblastic leukemia. Normal expression of TAL1 is observed in erythrocytic, megakaryocytic and mastocytic cells of the hematopoietic lineage. We now report that both RNA transcripts and polypeptide products of TAL1 are present in human umbilical vein endothelial cells cultured in vitro. Moreover, in situ hybridization revealed a restricted pattern of TAL1 expression in endothelial cells in vivo, including vessels within the white pulp and follicles of the spleen. In view of its presumptive role as a transcriptional factor, the TAL1 gene product may serve during normal development as a regulator of endothelial cell growth or differentiation. PMID- 8414505 TI - Hepatocyte growth factor transforms immortalized mouse liver epithelial cells. AB - Transforming activity of hepatocyte growth factor (HGF) was demonstrated utilizing immortalized but not fully transformed mouse hepatocytes (MLE-10). Rat HGF cDNA, expressed under the control of a cytomegalovirus promoter, was transfected together with the neomycin resistance gene (PSV2neo) into MLE-10 cells by the calcium phosphate method, and propagated G418-resistant colonies were harvested colony by colony. After checking for integration and expression of exogenous HGF, five cell lines (MLE-10-HGF-1-5) were established. Three cell lines transfected with the vector only (MLE-10-CMV-1-3) were also established in the same manner. All MLE-10-HGF cell lines grew much faster than the MLE-10-CMV and original MLE-10 cells in culture and produced large colonies in soft agar, which colony production was blocked by the addition of anti-HGF antibody to the agar. After addition of HGF, original and MLE-10-CMV lines produced colonies in soft agar. The high-HGF-production lines (MLE-10-HGF-4 and -5) also gave rise to tumors within 2 weeks when implanted into the nude mice subcutis. In contrast, all MLE-10-CMV and original MLE-10 cells were negative in these growth assays. A rough parallelism between the level of HGF expression and the growth rate in both soft agar and nude mice subcutis was evident among MLE-10-HGF cell lines. Those with higher HGF production tended to grow in a scattered fashion in culture. High affinity HGF receptor, HGFR/met, was expressed in MLE-10 and all the derived cell lines. Since HGF and/or HGFR/met gene expression is seen in various tumors and the serum HGF level is elevated in patients with hepatic disease, the present results indicate a possible significance of HGF and its receptor system in carcinogenesis, most probably via autocrine and/or paracrine mechanisms. PMID- 8414506 TI - Increased sequence-specific p53-DNA binding activity after DNA damage is attenuated by phorbol esters. AB - Damage to cellular DNA greatly increases the levels of the tumor-suppressor gene p53 and induces cell cycle arrest in G1. A critical function of wild-type p53 is its ability to bind to specific DNA sequences. The effect of DNA damage on the sequence-specific DNA-binding properties of cellular p53 was investigated using DNA gel mobility-shift assays with nuclear extracts from NIH-3T3 cells. DNA damage (initiated by radiation) induced a rapid, cycloheximide-sensitive increase in the levels of nuclear p53-DNA binding activity and an increase in the half life of the p53 protein. Increased p53-DNA binding activity could be detected at low (0.2 Gy), non-lethal doses of radiation. The tumor promoter 12-O tetradecanoyl phorbol 13-acetate (TPA) attenuated the DNA damage-induced increase in p53-DNA binding activity by decreasing the half-life of the p53 protein. The tumor promoter properties of TPA may therefore be mediated by interfering with the cellular p53 response to DNA damage. The increased levels of p53 bound to specific DNA sequences following DNA damage may induce cell cycle arrest. p53 mediated growth arrest could occur by inhibition of DNA replication and/or alterations in transcription of cell cycle genes. PMID- 8414507 TI - Molecular events including p53 and k-ras alterations in the in vitro progression of a human colorectal adenoma cell line to an adenocarcinoma. AB - The aim of the current study was to identify genetic abnormalities in human colorectal adenoma and carcinoma derived cell lines, and to determine whether the genetic changes which occur in vitro are relevant to the in vivo situation. Loss of 1p(33-35) region was shown to be the most common chromosome 1 abnormality and loss of heterozygosity (LOH) of the DCC gene and/or adjacent sequences was detected in all adenoma derived cells as well as the carcinoma cell lines. The level of p53 protein was also investigated as increased cellular p53 protein had previously been associated with mutation of the p53 gene. A further aim was to investigate genetic changes in our in vitro model of tumour progression, where the adenoma derived PC/AA cell line has previously been converted in vitro to two distinct tumorigenic phenotypes, producing either an adenocarcinoma or a mucinous carcinoma in athymic nude mice. Progression to the adenocarcinoma phenotype was shown to involve a specific chromosome 1 rearrangement, loss of both normal copies of chromosome 18 (although DCC gene sequences were retained), loss of the remaining wild type allele of k-ras resulting in homozygosity for the k-ras codon 12 mutation and increased cellular p53 protein as detected by SDS-PAGE Western blotting. The increase in p53 protein was shown not to be due to the acquisition of a mutation in the p53 gene. Interestingly, progression of the adenoma derived PC/AA cell line to the mucinous malignant phenotype did not involve any of these molecular rearrangements, suggesting that different genetically distinct pathways are involved in colorectal carcinogenesis. These studies show that the genetic changes in our in vitro model of human colorectal tumour progression are similar to those observed in in vivo studies. PMID- 8414508 TI - BCL2 complex rearrangement in follicular lymphoma: translocation mbr/JH and deletion in the vcr region of the same BCL2 allele. AB - Two rearrangements affecting the same allele of the BCL2 gene were characterized by molecular analysis of an untreated follicular lymphoma. The first rearrangement interested the major breakpoint region (mbr) on chromosome 18 and a JH segment on chromosome 14. The other one was located at the 5' end of the BCL2 gene, in the so called variant cluster region (vcr), and consisted of a series of deletions that removed part of a DNA region where initiation of transcription normally occurs. Interestingly, both rearrangements involved the same BCL2 allele. The simultaneous presence of mbr (or mcr) translocations and of minor rearrangements in vcr has been previously suggested by restriction map analysis in a significant number of follicular lymphomas. The significance of these abnormalities on the oncogenic process is discussed. PMID- 8414509 TI - The catalytic domain of the mouse sos1 gene product activates Ras proteins in vivo and in vitro. AB - Nucleotide exchange factors (NEFs) which are structurally related to the yeast Saccharomyces cerevisiae CDC25 gene product have recently been identified in mammals. One of these NEFs, cdc25, has been shown to activate RAS in yeast and to promote nucleotide exchange on RAS proteins in vitro. The cdc25 from mammals is expressed at high levels in brain tissue but not in a variety of other tissues examined. The vertebrate sos1 and sos2 gene products have a domain structurally related to the catalytic domain of the yeast CDC25NEF. The expression pattern of sos1 and sos2 is widespread, showing detectable levels of expression in all tissues examined, although the levels vary dramatically in various tissues. In this report we demonstrate that the catalytic domain of SOS1NEF can complement the loss of CDC25 function in yeast, can bind tightly to the nucleotide-free form of H-ras in vitro and can promote nucleotide exchange on the H-ras protein in vitro. PMID- 8414510 TI - MLLT3 gene on 9p22 involved in t(9;11) leukemia encodes a serine/proline rich protein homologous to MLLT1 on 19p13. AB - Recently, the MLL gene at 11q23 was found to be involved in a subset of leukemias with an 11q23 abnormality. In the present study, we isolated chimeric cDNAs between the MLL and a gene designated MLLT3 at 9p22 from a cDNA library of an IMS M1 cell line with a t(9;11)(p22;q23) translocation, a representative karyotypic abnormality seen in acute monocytic leukemia. We also isolated a normal MLLT3 cDNA and found an open reading frame encoding at least 318 amino acids with high serine/proline content (24.8%). The chimeric mRNAs were demonstrated to be fused to MLL in frame, as found in t(11;19) and t(4;11) leukemias. The predicted MLLT3 protein demonstrated a significant homology to that of the MLLT1 gene at 19p13 involved in t(11;19) leukemia. The highest homology, up to 74.1%, was found in 86 amino acids of the C-terminus, suggesting that this region is of particular importance for leukemogenesis in t(9;11) leukemia. Northern blot analysis with the MLLT3 cDNA probe against normal tissues revealed multiple transcripts in lymphoid organs. A survey of hematopoietic cell lines demonstrated relatively stronger signals in cells belonging to megakaryocytic and erythroid lineages. As previously found in t(11;19) leukemia, heterogeneous MLL-MLLT3 chimeric mRNAs could be detected by the reverse transcriptase-polymerase chain reaction (RT-PCR) in t(9;11) leukemia samples. PMID- 8414511 TI - SCL, the gene implicated in human T-cell leukaemia, is oncogenic in a murine T lymphocyte cell line. AB - SCL (TAL-1) is implicated in the generation of human T-cell acute lymphoblastic leukaemia. To directly examine the role of this putative oncogene, an SCL retrovirus was constructed and used to infect a v-ABL transformed T-lymphocyte cell line. Thirteen independent SCL-infected and four control cell lines were established and injected subcutaneously into syngeneic mice. Mice injected with SCL-infected clonal cell lines died significantly more rapidly than control animals. By day 200 46% (40/87) of animals injected with SCL-infected cell lines had died due to disseminated transplantable lymphoid tumours. In contrast only 22% of control mice were dead by day 200 (P < 0.0015). Of possible relevance to the enhanced tumourigenesis, some SCL-infected cell lines displayed increased clonogenicity in agar. Increased cell growth was even more striking when ex-vivo tumour-derived cell lines were studied. Thus, SCL can co-operate with v-ABL to hasten T-cell tumourigenesis. This is the first direct evidence demonstrating that SCL can behave as an oncogene. PMID- 8414512 TI - Overexpression of c-erbA proto-oncogene enhances myogenic differentiation. AB - Triiodothyronine (T3) positively regulates both the expression of the MyoD gene, a key myogenic regulator, and C2 muscle cell differentiation. To directly examine the role of its nuclear receptors in the control of myogenesis, we introduced a c erbA expression vector into C2 muscle cells by transient or stable transfection. Our results show that c-erbA can play a potent role in the triggering of muscle terminal differentiation since its overexpression leads to: (1) a complete abrogation of the activity of the myogenesis inhibitor AP-1 (fos/jun) transcription factor; (2) an enhanced induction of MyoD expression upon T3 treatment; (3) the acquisition by T3 of the ability to trigger both growth arrest and terminal differentiation in the presence of large amounts of serum mitogens, a property that is otherwise specific to retinoic acid (RA). Thus, c-erbA is one of the two protooncogenes (with c-ski) that acts as positive regulator of muscle differentiation. Furthermore, the fact that c-erbA overexpression allows T3 to largely mimic the RA effects indicates that their biological differences in the modulation of myogenic program primarily rely on the differential expression of their receptors in C2 muscle cells rather than on an intrinsic specificity of their target genes. PMID- 8414513 TI - Retroviral enhancer insertion 5' of c-myc in two translocation-negative mouse plasmacytomas upregulates c-myc expression to different extents. AB - Essentially all murine plasmacytomas have deregulated c-myc expression that is typically brought about by chromosomal translocations between the c-myc/Pvt-1 locus and one of the immunoglobulin loci. ABPC 22 and RFPC 2782 are BALB/c plasmacytomas that lack chromosomal translocations yet have Southern blot evidence of c-myc gene rearrangements. In this report we show that proviral integrations 5' of the c-myc gene can deregulate c-myc expression in mouse plasmacytomas. Analysis of DNA sequences 5' of the c-myc genes from both tumors demonstrated that rearrangements were caused by retroviral integrations 5' of c myc exon 1. The proviral insertion in RFPC 2782 was associated with a high steady state c-myc mRNA level comparable to that seen in plasmacytomas with typical translocations. An analogous proviral insertion in ABPC 22 was associated with a c-myc RNA level that was only 38% of that of RFPC 2782. Nuclear run-on studies of c-myc transcription showed that ABPC 22 has both a lower rate of transcription and a greater degree of transcriptional attenuation than RFPC 2782. DNA sequencing of the long terminal repeat of each tumor provirus showed that the ABPC 22 provirus harbors a deletion of one of the two direct repeats in the viral enhancer, whereas both repeats are present in the RFPC 2782 provirus. These data indicate that maximum LTR enhancer effectiveness in plasmacytomas in vivo requires the presence of both LTR direct repeats. The documentation of the low level of steady-state c-myc mRNA in ABPC 22 supports the notion that deregulated c-myc expression, even at low steady state levels, is effective in supporting the development of plasmacytomas. PMID- 8414514 TI - Characterization of the transcriptional regulatory region of the human WT1 gene. AB - Specific control of the expression of the Wilms tumor gene WT1 is important for normal development of the kidney. In order to characterise the transcriptional control region of the WT1 gene we have isolated genomic clones spanning the upstream region, the first WT1 exon and the 5' end of the Wit1 gene. DNA sequencing revealed that the WT1 promoter lacks a TATA box or CCAAT motif and has a GC content of 71%. Four transcriptional start sites are clustered within a 32 bp region. GC-boxes are present at nucleotide positions -413, -160, +84 and +158. DNAase I protection assays with purified Sp1 protein revealed the existence of 11 different binding sites in the WT1 promoter. WT1 and Wit1 promoter activities were tested in COS-7 cells with luciferase reporter gene constructs either containing or lacking an SV40 enhancer. WT1 promoter activity was found in a fragment extending from 449 bp upstream to 201 bp downstream of the WT1 start site. It was 26 fold lower in the absence of the SV40 enhancer than in the presence. Cotransfection with a Sp1 expression vector stimulated both constructs 3-4 fold. Wit1 promoter activity was identified in a DNA fragment extending from 200 bp upstream of the putative Wit1 TATA box to 130 bp downstream. Several potential recognition sites for WT1/EGR, Pax-8, and GAGA-like transcription factors are present in the WT1 promoter. PMID- 8414515 TI - Redox regulation of a src family protein tyrosine kinase p56lck in T cells. AB - Protein tyrosine phosphorylation was examined after T cells were exposed to oxidative stress in vitro to investigate the possible involvement of redox regulation in T-cell signaling. Oxidative reagents such as hydrogen peroxide (H2O2) and diamide, which oxidize the free sulfhydryl groups in the cells, markedly induced tyrosine phosphorylation of multiple cellular proteins, especially a 55-kDa protein, of cultured peripheral blood T lymphocytes (PBL blasts). The 55-kDa molecule phosphorylated by diamide turned out to be a src family protein tyrosine kinase, p56lck. The immune complex kinase assay showed that the kinase activity of p56lck of diamide-treated PBL blasts was enhanced. The tryptic peptide mapping of p56lck demonstrated that diamide induced the phosphorylation both at Tyr-394 (autophosphorylation site) and at Tyr-505 (negative regulatory site). Taken together, the tyrosine phosphorylation and presumably kinase activity of p56lck were swiftly enhanced by oxidative stress, indicating that T cells have a redox-sensitive signaling mechanism, which is partly mediated by the lymphocyte-specific protein tyrosine kinase p56lck. PMID- 8414516 TI - Modulation of epidermal growth factor receptor activity and related responses by the 7-deazaguanine derivative, queuine. AB - Epidermal growth factor (EGF) induces autophosphorylation of its cognate receptor at tyrosine residues. Here we show that queuine (q), a widely distributed modified guanine analogue occurring free or as a tRNA wobble base, modulates this EGF receptor activity in vitro and in intact cells. Autophosphorylation of the immunopurified receptor from human A431 epidermoid carcinoma cells was enhanced three to fourfold in the presence of physiological concentrations of q. Using a membrane fraction of A431 cells, a twofold increase in autophosphorylation activity in the presence of q was observed, however, only when the receptor was activated by the ligand. In intact A431 cells, q enhanced the initial ligand induced autophosphorylation of the EGF receptor three to fourfold. However, upon longer treatment of the cells with EGF in the presence of q, significantly less autophosphorylated receptor was detectable compared with stimulation of cells in the absence of q. A similar q-dependent modulation of EGF receptor autophosphorylation was observed also in human cervical carcinoma cells HeLa-S3. Treatment of q-deficient HeLa cells with EGF induced the c-fos gene expression, transiently increased the activity of the anoxic stress protein LDH k, and stimulated proliferation. Treatment of HeLa cells with EGF in the presence of q resulted in a delayed c-fos gene expression and an accelerated increase and decrease of LDH k activity. The stimulatory effect of low doses of EGF on HeLa cell proliferation was completely antagonized in the presence of q. The results suggest that the mitogenic signalling initiated by the EGF receptor is modulated by q. PMID- 8414517 TI - Characterization of the retinoblastoma binding proteins RBP1 and RBP2. AB - The retinoblastoma gene product, pRB, regulates cell proliferation by binding to and inhibiting the activity of key growth promoting proteins. Several cellular proteins have been shown to bind directly to pRB and the genes encoding a number of them have been isolated. The protein product of one of these genes is the transcription factor E2F. We have now isolated cDNA clones that contain the full length coding sequence of two other proteins, RBP1 and RBP2, cloned originally by their interaction with pRB. The products of the RBP1 and RBP2 genes are ubiquitously expressed, large (200 kDa for RBP1 and 195 kDa for RBP2) nuclear phosphoproteins with structural motifs that suggest a role in transcriptional regulation. In addition we have been able to identify complexes of pRB and RBP1 in vivo that are dissociated in the presence of purified human papillomavirus E7 protein. PMID- 8414518 TI - A method for identifying genes within yeast artificial chromosomes: application to isolation of MLL fusion cDNAs from acute leukaemia translocations. AB - The majority of chromosomal translocations breakpoints are within regions of the genome where few DNA probes are available. The use of yeast artificial chromosomes (YACs) containing long stretches of human DNA allows dispersed DNA markers to be used to identify the position of breakpoints but does not readily allow subcloning of the precise breakpoint within the YAC DNA nor the cDNAs containing the affected genes. We describe a procedure allowing rapid isolation of cDNAs corresponding to genes within a YAC clone. Random cDNA is hybridised to PCR-generated biotinylated fragments of total DNA from a yeast strain harbouring a YAC clone. The hybrids can be recovered to facilitate subsequent cloning of the cDNA molecules. The application of this method to the cloning of cDNA molecules carrying sequences involved in the translocation t(4;11)(q21;q23) is illustrated. PMID- 8414519 TI - bmi-1 transgene induces lymphomas and collaborates with myc in tumorigenesis. AB - The bmi-1 gene was discovered as a frequent target of Moloney virus insertion in virally accelerated B-lymphoid tumors of E mu-myc transgenic mice and hence is thought to collaborate with the myc gene in lymphomagenesis, but its oncogenic potential has not previously been tested directly. To determine whether bmi-1 overexpression can contribute to hematopoietic neoplasia in vivo, strains of transgenic mice were generated in which bmi-1 expression was directed to the lymphoid compartment by a coupled immunoglobulin heavy chain enhancer (E mu). Although the E mu-bmi-1 transgene was expressed in both B and T cells, lymphoid development was not perturbed. Nevertheless, 14% of the mice in the strain with highest expression have developed lymphoma. Unexpectedly, most tumors were of the T-cell lineage, although one case of B lymphoma was observed. Furthermore, cross breeding E mu-bmi-1 and E mu-myc mice established that the bmi-1 transgene markedly accelerated the onset of pre-B and B lymphomas. These results demonstrate directly that bmi-1 can contribute to lymphomagenesis in the T and B cell lineages and collaborate with the myc gene in tumor development. PMID- 8414520 TI - Human p53 binds DNA as a protein homodimer but monomeric variants retain full transcription transactivation activity. AB - Wild-type human p53 protein is able to self-associate and consists predominantly of homotetramers in solution. In earlier work we identified the protein sequence motifs involved in p53 quaternary structure and showed that while monomeric p53 protein retains tumour suppressor function, monomeric tumour mutant p53 lacks dominant transforming activity. In this report we use point mutated and truncated cDNA genes encoding self-association defective human p53 proteins to investigate the relationship between p53 protein quaternary structure and the associated activities of transcription transactivation and target specific DNA binding. We show that p53 binds to a target oligonucleotide as a protein homodimer and that p53 dimerisation is required for detectable DNA binding. We found no evidence for p53 tetramer: DNA complexes and we suggest that the quaternary structure status of p53 may regulate a DNA binding associated activity. Monomeric p53 proteins failed to bind DNA in these assays but exhibited increased transactivating activity. Thus, both transcription transactivation and tumour suppressor functions act independently of p53 protein self-association and DNA binding. We propose that our results validate the p53 dimerisation motif as a target for rational anticancer drug design. We predict that compounds able to block p53 dimer assembly would inhibit the dominant transforming activities of mutant p53 in tumours retaining expression of a mutant allele, while leaving intact the wild type p53 associated activities of transcription transactivation and transformation suppression in unaffected tissue. PMID- 8414521 TI - Specific association of activated MAP kinase kinase kinase (Raf) with the plasma membranes of ras-transformed retinal cells. AB - Plasma membrane-enriched fractions were prepared from human embryonic retinal cells transformed with either adenovirus E1A and oncogenic ras DNA, or E1A and E1B DNA. Ras comprised 5-10% of the membrane protein from the E1A/ras transformed cells, whereas the membranes from E1A/E1B transformed cells did not overexpress Ras. The membranes from E1A/ras cells contained MAP kinase kinase kinase (MAPKKK) activity, even after washing in 0.5 M NaCl, whereas the membranes from E1A/E1B cells did not. Neither membrane fraction contained MAP kinase kinase or MAP kinase activity after washing with 0.5M NaCl. Immunoblotting experiments revealed about 10-fold more c-Raf in the membranes from E1A/ras cells than from E1A/E1B cells, and 50-60% of the MAPKKK activity in Triton X100-solubilised membranes from E1A/ras cells was immunoprecipitated with anti-Raf antibodies. A striking enrichment of c-Raf in the plasma membranes of E1A/ras cells was also demonstrated by immunocytochemistry, where it was co-localized with Ras. The MAPKKK activity in E1A/ras membranes was unaffected by incubation with protein phosphatases or by inclusion of protein phosphatase inhibitors during isolation, nor was it activated by GTP-Ras or inhibited by GDP-Ras. The results support the view that Ras and c-Raf interact with one another, but that neither c-Raf phosphorylation nor its interaction with GTP-Ras are alone sufficient for activation. The identification of MAPKKK activity in the membranes of ras transformed cells may prove useful in elucidating the mechanism by which Raf is activated by Ras. PMID- 8414522 TI - Ribozyme-mediated cleavage of the BCRABL oncogene transcript: in vitro cleavage of RNA and in vivo loss of P210 protein-kinase activity. AB - The t(9,22) chromosomal translocation generating the Philadelphia chromosome and the BCRABL oncogene has been shown both cytogenetically and molecularly to be the etiologic event in chronic myelogenous leukemia (CML). We have designed a ribozyme to cleave the BCRABL mRNA by targeting a GUU triplet adjacent to the junction of the c-BCR and c-ABL fused genes. This ribozyme efficiently cleaved BCRABL RNA transcripts as demonstrated by in vitro cleavage reactions. To determine the effect of constitutive expression of the ribozyme on the elimination of the BCRABL gene product, the ribozyme cDNA sequence was inserted into different retroviral expression vectors. Introduction of the recombinant retroviruses into the CML blast crisis cell-line K562, resulted in the elimination of the P210 protein-kinase activity in several single cell clones infected with the ribozyme expression cassette. Therefore BCR-ABL specific ribozymes may provide a potential genetic therapy for CML. PMID- 8414523 TI - What strategies have you used RN/MD relationships at your institution? PMID- 8414524 TI - Inflammatory diseases of the sinuses:anatomy. AB - Inflammatory sinus disease most commonly results from or is exacerbated by areas of obstruction within the sinuses. Recent revision of our understanding of mucociliary clearance has placed a renewed emphasis on sinus anatomy, particularly in identifying natural sinus ostia. A discussion of paranasal sinus anatomy is presented in this article along with special reference to these areas as well as contiguous sites that may be involved as a complication of inflammatory disease. PMID- 8414525 TI - Inflammatory diseases of the sinuses:physiology. Clinical applications. AB - Physiologic principles form the foundation upon which rational medical and surgical decisions are based. Over the years, a growing understanding of the physiology of the nose and sinuses has evolved, allowing us to better treat our patients with sinonasal disease. As clinicians, we are challenged to keep abreast not only of advances in therapeutic options but also of advances in physiology. In this manner, we may employ the new therapies with both skill and intelligence. PMID- 8414526 TI - Imaging of inflammatory sinus disease. AB - Sinusitis, especially as it relates to allergies, is one of the most commonly overlooked and misunderstood diseases in clinical practice. This article explores the radiographic appearance and anatomic characteristics that must be considered in the diagnosis and treatment of sinusitis. CT scans and MR imaging also are discussed in relation to the diagnosis of this disease. PMID- 8414527 TI - Inflammatory diseases of the sinuses: bacteriology and antibiotics. AB - This article provides an overview of the antimicrobial agents most commonly employed against the bacteria that cause infections in the ear, nose, throat, head, and neck. Because new bacterial resistances appear so regularly, as do new antibiotics, the reader is encouraged to supplement the information provided within this article with current information from the available literature. Specific treatment strategies for both acute and chronic sinusitis also are reviewed. PMID- 8414528 TI - Inflammatory diseases of the sinuses: an overview of immunology. AB - We have provided an overview of the immune system and its components. These components--cell-mediated and humoral immunity, the complement system and the different phagocytic cells--all function in concert to protect the body from foreign invaders. When this powerful immune system behaves in a misdirected fashion, however, disease states develop from its injurious effects. One such disease process, intimately related to sinus disease, is allergic rhinitis, which starts as an IgE-mediated response and evolves into an extensive reaction with the contribution of several inflammatory cells and mediators (Fig. 5). PMID- 8414529 TI - Endoscopic sinus surgery. AB - Successful endoscopic sinus surgery requires a thorough knowledge of the anatomy of the paranasal sinuses and, in particular, the lateral nasal wall. This article discusses a variety of topics relating to endoscopic sinus surgery, including physiology, patient evaluation, several surgical techniques, postoperative care, and complications. PMID- 8414530 TI - Chronic frontal sinus disease. AB - A variety of procedures are now available to manage chronic frontal sinus diseases. The choice of procedures depends upon both the extent of the disease and the experience of the surgeon. PMID- 8414531 TI - Pediatric sinusitis. AB - Interest in pediatric sinusitis is growing in response to better diagnostic techniques and to the implication of better results with newer surgical techniques. In addition, sinusitis has the potential for a significant impact on overall health in growing numbers of children with chronic pulmonary disease and immunodeficiency. PMID- 8414532 TI - Inflammatory sinus diseases affecting the orbit. AB - Paranasal infections and inflammatory processes involving the orbits have been described, as have primary orbital conditions that may mimic acute sinus disease. Most common are bacterial and fungal sinus infections with secondary orbital spread, diseases for which the otolaryngologist-head and neck surgeon must be skilled in diagnosis and management in order to minimize sequelae. Optic neuritis arising from such infections requires prompt recognition and aggressive treatment if vision is to be preserved. The most common causes of unilateral proptosis, Graves' orbitopathy, and pseudotumor may in their early stages mimic orbital spread of paranasal infections. Recognition of these entities and ophthalmologic consultation are critical to their appropriate management. PMID- 8414533 TI - Surgery of the anterior skull base. AB - Inflammatory disease of the frontal, ethmoid, and sphenoid paranasal sinuses may extend to the adjacent anterior skull base and then intracranially. The potential for this serious complication of sinus disease must be recognized by the primary otolaryngologist-head and neck surgeon, and alternative management strategies must be taken once it is diagnosed. Topics discussed in this article include cranial anatomy, surgical techniques and variations, complications, and special considerations. PMID- 8414534 TI - Difficult decisions in endoscopic sinus surgery. AB - In this article, case studies from four separate clinicians are presented along with comments. These cases offer unusual examples of problems that occur with endoscopic sinus surgery. PMID- 8414535 TI - [Magnetic resonance imaging in otolaryngology]. AB - In the paper authors describe fundamental physical properties of a phenomenon of the radio-frequency excitation and relaxation of nuclei ordered in a strong magnetic field and the usefulness of MRI in medical diagnostic procedures. Basic interpretations principles of MR imaging due to signal intensity differences between organs and tissues in T1- and T2- weighted sequences and proton density are presented. Both, literature review and experience of authors suggest application of MRI in otolaryngology, it is illustrated by a lot of examples. The MR imaging studies were compared with results obtained from CT in otolaryngology field. PMID- 8414536 TI - [Prognostic factors in case of metastases to the cervical lymph nodes of the laryngeal cancer]. AB - Affection of the elements of the lymphatic system in the cancer of the larynx or hypopharynx is making the prognosis worse. We know that this is the simplification of the problem. We are trying to find and define factors which have the main meaning in prognosis of the treatment and the survival of the treated patients. In order to attain this we analyse histopathological material and results of clinic observations. PMID- 8414537 TI - [Laser aryntenoidectomy in bilateral vocal cord paralysis treatment]. AB - The method of surgical treatment of bilateral vocal cord paralysis with the use of laser CO2 is presented. This technique was used in 20 patients. The vaporization and extirpation of arytenoid cartilage was performed together with resection of 1/3 of the posterior vocal cord which was removed. Analyzing other methods of treatment of bilateral vocal cord paralysis it was emphasized that with the use of the presented method good results in preserving breathing and phonation of the operated part was archived. It was also emphasized that the fast process of healing and lack of granulation in the operated field allows for the quick recovering patient. This method allows to obtain good wide larynx lumen without the necessity to perform tracheotomy--in 17 out of 20 our patients we could perform laser arytenoidectomy without tracheotomy. PMID- 8414538 TI - [The diagnostic value of Magnetic Resonance in otolaryngological diseases]. PMID- 8414539 TI - [The results of treatment of juvenile laryngeal papillomatosis in the light of experience of the ENT department in Krakow]. AB - In the years 1973-1990, 48 children with laryngeal papillomatosis were treated in the Department of Otolaryngology, Medical University of Cracow. The data was obtained in 42 children. In 51% children the entire and long-lasting (from 4 to 8 years) remission of papillomatosis was ascertained. In 39% patients the considerably limited papillomatosis with distinctly prolonged remission time was obtained and in 9% of the patients the development of papillomatosis in low respiratory tract was noted. PMID- 8414540 TI - [Smell and taste in patients with neoplastic tumors of palatine tonsils treated by radiation]. AB - The smell and taste investigations were made in group of 47 subjects with neoplasms tumors of palatine tonsils surgical treated with radiation and in group of 7 subjects treated only by radiation. The investigation presents mostly quantitative disturbance of smell in both groups but with temporary character. PMID- 8414541 TI - [Endoscopic head: the ultrasound examination of mouth, pharynx and nasopharynx]. AB - The objective evaluation of the character of a change, localized in the area of palatine tonsil and tongue, especially in case of defining the extension of the change may cause many problems. So far the ultrasound examination of the tongue and tonsil was carried out by way of placing the endoscopic head on the skin in submandibular area or just behind the branch of mandibular, which in many cases didn't allow to achieve a satisfactory image of the changed tissues. This paper presents an attempt to use endoscopic head to examine changes localized in the area of tongue, palatine tonsil and nasopharynx and on soft palate. The paper also evaluates the usefulness of 2 types of endoscopic heads to carry out such examinations. The obtained initial results of the examinations point at substantial usefulness of this type of examination to the evaluation of changes localized in the area of tongue and palatine tonsil. PMID- 8414542 TI - [The role of haemophilus in respiratory tract diseases in children from Silesian Center of Children Rehabilitation in Rabka]. AB - In this work the frequency of isolation of rods the genus Haemophilus from children suffering from various clinical sharpes of respiratory tract diseases. All isolated strains were tested in respect of species, biotype, the ability of adhesion to oral and laryngeal epithelia and their susceptibility to routinely applied antibiotics. A strong correlation between the species and biotype of rods from the genus Haemophilus and clinical shape of respiratory tract diseases was found. It was observed that the Haemophilus rods show differences in the ability of adhesion to oral and laryngeal epithelia. Only 52% of the isolated strains were susceptible to bactrim and 88% to tetracycline. PMID- 8414543 TI - [Sonographic imaging of internal jugular venous blood flow in juvenile hyperfunctional dysphonia]. AB - Relative blood flow velocity peaks of the internal jugular vein were examined by simultaneous Doppler and B-mode sonography in 4 boys with hyperfunctional dysphonia. During phonation patients showed remarkably greater reduction of relative blood flow velocity peaks than a normal speaker. PMID- 8414544 TI - [A case of rhabdomyosarcoma of trachea in a 7-year-old child]. PMID- 8414545 TI - [Congenital nasal lipoma]. AB - The authors describe a rare case of congenital lipoma of a nose and nasopharynx in a 7 months old girl. The tumor, about 7 x 2 cm, was situated in the right nasal cavity and the nasopharynx. The tumor caused complete obstruction of the right side of the nose. The tumor was excised from intranasal approach. Histological examination disclosed lipoma. The duration of follow up was 10 months without any sight of recurrence. PMID- 8414546 TI - [Salivary gland lipoma]. PMID- 8414547 TI - [The exceptional way of spreading of the parapharyngeal space abscess]. AB - The authors described the exceptional way of spreading of the parapharyngeal space abscess. Succession of this process were hypostatic abscesses of the lumbo sacral region. Incision and drainage parapharyngeal abscess and as well hypostatic abscesses pointed out the usefulness in treatment. PMID- 8414548 TI - [Extensive cervical emphysema as a result of laryngeal injury]. AB - The authors describe a case of rare larynx injury in the subglottic region. A fracture of cricoid cartilage caused a presence of air in prevertebral and perithyroid space. The diagnosis has been established on the ground of conventional x-ray examination. PMID- 8414549 TI - [Castleman's disease: an unusual neck neoplasm]. AB - Castleman's tumour of unusual location on the neck of 12 years old girl has been described. From two forms of Castleman's disease which exist--the hyaline vascular kind of disease. The authors presented their opinion on the affection arising and differential diagnosis Castleman's tumour from tumours of thymus gland origin. PMID- 8414550 TI - [Difficulties in histopathologic diagnosis of the metastases of the renal clear cell carcinoma to the nasal cavity]. PMID- 8414551 TI - [Investigations and evaluation of occupational exposure of the upper respiratory tract on the derivatives of the petroleum at employees in petrochemical industry]. PMID- 8414552 TI - Heinrich Neumann: multifaceted otologic surgeon. PMID- 8414553 TI - Patterns of hearing loss resulting from cis-platinum therapy. AB - The purpose of this study was to provide a clinically useful classification of the hearing loss that occurs with chemotherapy using cis-diamminedichloroplatinum (cis-platinum). Serial audiometric testing was performed on 66 patients who underwent cis-platinum chemotherapy (2 to 6 courses, 100 mg/m2). Subsequent hearing losses were classified according to (1) whether hearing loss occurred after the first or second course of chemotherapy, (2) the magnitude of loss, and (3) the frequencies affected. Of 39 patients who could be classified, 21 (54%) had no loss or mild loss, 14 (36%) had early loss, and 4 (10%) had late loss. Different patterns of hearing loss were observed. The observation that different patterns of hearing loss occur implies that different mechanisms of hearing loss exist. Some mechanisms must be more important in different patients. Our data are important with regard to continuation of chemotherapy treatment after hearing loss occurs. Most significant hearing loss (90%) occurred early or not at all. If early hearing loss occurred and treatment was continued, the speech frequencies were eventually affected in 71% of patients. Recognition of the patterns of hearing loss are important considerations when discontinuation of therapy is being considered. PMID- 8414554 TI - Evaluation of patients with acoustic neuroma with dynamic posturography. AB - The purpose of the present work was to consider the use of posture tests in patients who had known acoustic tumors. The results suggest that it might be possible to determine whether the tumor is on the inferior or superior branch of the vestibular nerve. This would have clinical significance for planning a surgical approach to the tumor. PMID- 8414555 TI - Autorotation test of the horizontal vestibulo-ocular reflex in Meniere's disease. AB - Sixty-four patients with the diagnosis of Meniere's disease were tested at the House Ear Clinic with an active head-rotation test system, the Vestibular Autorotation Test (VAT). The VAT is a portable, computerized test that measures the horizontal vestibulo-ocular reflex (VOR) with the use of high-frequency (2 to 6 Hz) active head movements to obtain gain and phase. The purpose of this study was to characterize the horizontal VOR at high frequencies in patients with Meniere's disease. At frequencies from 5 to 6 Hz, all patients demonstrated horizontal phase greater than 180 degrees and 85% showed abnormal VAT results. The most common patterns were decreased gain or increased phase values, or both, relative to normative data. No significant differences in the degree of abnormality in gain and phase were noted among groups of patients when the patients were clinically staged. We conclude that, in our test population of patients with Meniere's disease, the VAT shows common gain and phase patterns and abnormalities of the horizontal VOR. This may contribute to high retinal image velocities, which render the patient unable to stabilize retinal images during locomotion (visual field image slip), in as many as 85% of the patients tested, regardless of clinical stage. Such high-frequency testing can reveal abnormalities of the horizontal VOR not apparent from conventional vestibular testing. Thus VAT provides additional information about the functioning VOR when combined with the present vestibular test battery. PMID- 8414556 TI - Pathogenesis of tympanosclerosis. AB - In spite of the wealth of information on the clinical, histologic, and pathologic aspects of tympanosclerosis, the pathogenesis of tympanosclerosis is still unclear. In an attempt to understand the pathogenesis, 319 human temporal bones from 196 individuals with otitis media were studied. The extent and nature of tympanosclerosis and the characteristics of the otitis media associated with it were studied. Forty-five temporal bones from 35 individuals with otitis media were found to have tympanosclerosis, giving an incidence of 14.1%. It was seen most commonly in individuals over 40 years of age (86.7%). The male-to-female ratio was 1.6:1. The most common site of occurrence was the tympanic membrane (88.9%). Tympanosclerosis was seen more often in the anterior and posterior inferior quadrants of the tympanic membrane and that, too, in a central position. Tympanosclerosis was seen more commonly in temporal bones with irreversible inflammatory changes, and in this group, late plaques were more commonly seen than early or intermediate plaques. Audiometric charts failed to show any direct relationship between extent of tympanosclerosis and the severity of hearing loss. The only audiometric finding of any consequence was a mixed hearing loss in the presence of middle ear tympanosclerosis. PMID- 8414557 TI - Is the vestibular system affected by middle ear effusion? AB - The association between middle ear effusion (MEE) and vestibular pathology is controversial. To investigate this point, 50 children with MEE scheduled for myringotomy and grommet, and 20 normal hearing children without MEE scheduled for adenotonsillectomy, underwent vestibular investigations by craniocorpography and rotatory chair, preoperatively and postoperatively. Most of the correlations, preoperative vs. postoperative, and study vs. control groups, were nonsignificant. Assuming that serous labyrinthitis is responsible for vestibular involvement in MEE, the lack of significant vestibular pathology in our study could probably be explained by the fact that no children with acute otitis media episodes or otalgia were included. PMID- 8414558 TI - Mycobacterial infections of the head and neck. AB - Despite the decline of pulmonary tuberculosis in the Western world, the incidence of cervical mycobacterial infections has remained relatively unaffected. A retrospective review was conducted of 20 patients treated for head and neck tuberculosis from 1984 to 1991. They were mostly an immigrant group coming from underdeveloped countries close to the equator. All cases were treated with antituberculous chemotherapy; 18 patients also underwent surgical excision of their lymphadenopathy. Sixteen patients showed complete response to combined treatment and one relapse was successfully retreated with antituberculous medication. Three patients died from unrelated causes--two from human immunodeficiency virus infection and one from nasopharyngeal carcinoma. The most reliable indicator of mycobacterial infection was the pathologic specimen, making the need for surgical intervention an important diagnostic consideration. As with other reports, most patients in our series had negative chest x-ray films, variable responses to skin testing, and negative cultures. The epidemiology, pathogenesis, and management of this disease are discussed. PMID- 8414559 TI - The surgical workstation: surgical planning using generic software. AB - Computer software for rendering and display of three-dimensional data is becoming readily available for all types of computers. Such programs typically accept data from any source, compute a three-dimensional volume of data, and display it with a variety of rendering options. Although not specifically designed for medical image processing, these programs can provide very detailed and finely rendered images that are useful for surgical planning. We use one such program to display data from standard computed tomography scans, which gives us a photorealistic three-dimensional view of patient anatomy. This view can be modified to render tissues transparent, translucent, or opaque, and thus allows the surgeon to selectively enhance bony architecture, tumors, or other details. Images can be rotated, sliced, and displayed in the surgical position. Image animation can be added to facilitate the display of complex anatomic relationships. Our experience with this technology suggests that such programs can provide the basis for personal surgical workstations for medical image analysis and surgical planning. Further development of such generic imaging systems should allow this useful technology to become widely available for surgical planning and education. We discuss our experience with a typical generic imaging workstation. PMID- 8414560 TI - Acute laryngeal trauma: a review of 77 patients. AB - Acute laryngeal trauma is a rare injury. In the past 18 years, 77 patients with acute laryngeal trauma have been evaluated at our institution. Each patient's care was overseen by the senior author (E.S.P.). The 61 patients who were seen within 48 hours of their accident are compared with those treated after 48 hours. All patients are classified by both injury (groups 1 through 5) and treatment (types I through III). Results are reported for voice, airway, and swallowing. Our methods of evaluation and treatment are outlined, and controversial aspects of patient management are addressed. We conclude that conservative treatment of group 1 and 2 injuries is 100% effective, expeditious repair of laryngeal injuries greatly reduces poor outcome, and the type of injury can be used to roughly predict patient outcome. Further, with use of current methods of diagnosis and management, almost all patients will be decannulated (98%) with functional speech (100%) and normal deglutition (100%). PMID- 8414561 TI - Intranasal histamine challenge in normality and allergic rhinitis. AB - A series of investigations was performed in which histamine challenge was used to compare nasal responsiveness in 20 normal subjects and 20 with allergic rhinitis. There was found to be a lower threshold of reactivity (D100) to histamine in allergic subjects as measured by resistance changes (geometric mean, 0.53 mg/ml; normal subjects, 2.15: p = 0.022). This may represent increased number or sensitivity of histamine receptors on the nasal capacitance vessels. The loss of a laser Doppler response to a supramaximal histamine stimulus (normal subjects, 102% increase in flux at 3 minutes; p < 0.05) was observed in patients with allergic rhinitis and indicates either a down-regulation of the capillary system or an altered effect of histamine on superficial vessels, perhaps mediated by a shift in histamine receptor type. There was an observed increase in neutrophils at the mucosal surface under baseline conditions (rhinitis median, 49.6%; normal subjects, 32.72%: p < 0.05), which suggests an important primary role in the pathogenesis of this condition for this active cell. The observed increase in secretory volume response to histamine in allergic subjects, which persisted beyond 40 minutes after a single D100 challenge, may be related to an altered sensitivity of glandular tissue. There are important changes in nasal reactivity to histamine challenge in allergic rhinitis that may have implications for its pathogenesis. PMID- 8414562 TI - Acute epiglottitis: changing epidemiologic patterns. AB - We determined incidence of acute epiglottitis in the Northern California Region of the Kaiser Permanente Medical Care Program during an 11-year period, from 1980 through 1990. The health plan membership is representative of the ethnicity, age, sex, and occupation of the Northern California population. The study included 135 children and 258 adults. Annual incidence in adults was relatively stable, ranging from 1.16 to 2.12 cases per 100,000. Annual incidence in children, however, decreased from 3.47 cases per 100,000 in 1980 to 0.63 cases per 100,000 in 1990. The ratio of annual incidence in children to that in adults decreased from 2.6 in 1980 to 0.4 in 1990. This changing epidemiologic pattern may be the effect of the Hemophilus influenzae type b vaccine (Hib), which was first given to children in 1985. Acute epiglottitis, classically considered a disease of children, is now becoming a disease of adults. PMID- 8414563 TI - The effects of etidronate disodium on progressive hearing loss from otosclerosis. AB - A 2-year prospective double-blind study was performed to evaluate the role of etidronate disodium for the treatment of progressive hearing loss in patients with otosclerosis. A pulsed dosage regimen was used during the 2-year period and the patients were followed up with otologic and audiometric examinations. Although statistically significant differences were not achieved between the study and control groups, the study did reveal a trend toward stabilization or improvement in air conduction thresholds in some frequencies (1000 and 4000 Hz) and in bone conduction thresholds at other frequencies (500, 1000, and 2000 Hz). The incidence of adverse side effects was similar in the treatment and control groups. Although no definite conclusions can be drawn from this pilot study, the findings provide encouragement for performing a larger and longer-term study. PMID- 8414564 TI - Management of laryngotracheal stenosis on the basis of site and severity. AB - Seventy-five cases of laryngotracheal stenosis treated between 1981 and 1991 were reviewed to determine the effectiveness of surgical treatment on the basis of site and severity of stenosis. Decannulation and absence of exertional dyspnea were the criteria of successful management. The treatment methods used were endoscopic laser incision and dilatation, expansion laryngotracheoplasty, and segmental resection. Endoscopic procedures were effective in treating thin (< 1 cm) stenoses in the subglottis and trachea. Laryngotracheoplasty was most effective in treating thick stenoses of the glottis and subglottis. Tracheal stenoses were most effectively treated by segmental resections. The probability for decannulation decreased with longer narrower stenoses and with increasing clinical stage. PMID- 8414565 TI - Adult supraglottitis. AB - Airway management of supraglottitis in the adult is controversial as regards the choice between observation and acute intervention. This study was undertaken to determine whether factors exist that place patients at higher risk of airway obstruction and to review conservative treatment outcome. Of 46 cases during an 8 year period, seven required tracheostomy and 39 were observed. Odynophagia and fever were present in all patients. Yearly clusters of up to six cases in 1 month were detected. White blood cell count and heart rate were significantly more elevated in those who underwent tracheostomy. A symptom duration of less than 24 hours was associated (p < 0.05, chi 2 analysis) with airway compromise as well. There was no associated morbidity or mortality, and mean hospital stay was 5.2 days for those observed and 7.9 days for those who underwent tracheostomy. PMID- 8414566 TI - Clinical guideline development for otitis media: a report on methodology. AB - Clinical guideline development is a major emphasis of recent health policy efforts. Interest in clinical guidelines is the result of multiple factors, including economic pressures and the desire to achieve a baseline level of practice in clinical settings. Guidelines have always been of fundamental importance in medicine. This article will discuss the place clinical guidelines have in contemporary practice and the difficulties encountered in the process of developing a meaningful guideline for managing otitis media with effusion. PMID- 8414567 TI - A comparison of growth rates of acoustic neuromas: nonsurgical patients vs. subtotal resection. AB - A conservative approach to the management of acoustic neuromas in elderly patients has been used since 1971. Elderly patients without symptoms of brain stem compression are initially treated by observation and yearly radiographic imaging. A translabyrinthine radical-subtotal resection is performed if brain stem compression is present or if tumor is growing rapidly. Twenty-three patients, ages 65 to 86 years, had initial nonsurgical management of their tumors. Growth rates could be determined for 16 patients. Thirteen patients not requiring surgery had an average tumor growth rate of 0.6 mm/yr. Three patients with an average growth rate of 6.8 mm/yr eventually required surgery. No patient whose tumor was < 15 mm at initial evaluation has experienced brain stem symptoms or demonstrated rapid tumor growth. Twenty-four patients ages 65 to 86 years underwent planned subtotal tumor excision. Eighteen patients followed postoperatively for more than 1 year demonstrated an average rate of regrowth of tumor of 0.7 mm/yr. PMID- 8414568 TI - Middle ear air injection after chronic ear surgery. AB - In the early period after chronic ear surgery, the reasons for conductive hearing loss may be difficult to determine. Patients who cannot autoinflate the middle ear after 3 weeks, or who have a negative Rinne test result with the 512 Hz tuning fork, are treated with a transtympanic injection of 0.5 cc of air with a 27-gauge needle and tuberculin syringe. This represents 20% of patients who had chronic ear surgery. Results show that hearing may be immediately improved, the sensation of pressure in the ear may be reduced, and fluid may be cleared from the middle ear. Other benefits may include the release of adhesions. The surgeon is better able to assess the thickness of the graft, and the status of the ossicular chain reconstruction can be determined. There have been no complications of middle ear infection or failure of the micropuncture site to heal. In our practice, middle ear air injection is a routine procedure in patients with inadequate eustachian tube function after chronic ear surgery. This report describes the results of 100 patients over 14 years who received middle ear air injections after chronic ear surgery compared with a control group of 100 patients who did not meet the criteria for requiring air injection. Hearing was immediately improved in 74% of patients as determined by Rinne testing. Audiograms were performed in 25 of these patients, documenting a mean improvement in pure-tone average of 16 dB. The long-term hearing results in patients undergoing air injection, who by definition had evidence of poor eustachian tube function, are similar to the results in the control group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414569 TI - Teflon vocal fold augmentation: failures and management in 28 cases. AB - Although vocal fold augmentation by Teflon injection has been the mainstay of treatment for glottic insufficiency for three decades, the success and safety of this treatment have been overstated. Twenty-eight patients who manifested poor or complicated Teflon results between 1984 and 1991 were evaluated using acoustic, aerodynamic, videostroboscopic, perceptual, and subjective patient self evaluation of voice, both before and after our management of these complications. Most of these had Teflon granulomas; subglottic overfilling was the most common condition. In most instances such management included microsurgical removal of the Teflon granuloma. Voice measures that were abnormal before correction tended to improve and move into the normal range, although the resultant voices were not totally normal. Degree of improvement varied depending on the Teflon-induced tissue changes and the methods of correction subsequently used. The worst results were in patients with scarring, atrophy, and bilaterally mobile vocal folds, for whom Teflon should never have been injected. Teflon injection should be reserved for those instances in which it is clearly indicated and the surgeon is skilled in the technique of intrafold injection. PMID- 8414570 TI - Injuries of the hard palate and the horizontal buttress of the midface. AB - Traumatic injuries to the hard palate occurred in 20% of all Le Fort midfacial fractures. The side of a palatal split was directly related to the side that received the highest Le Fort injury. In a symmetric midfacial injury, the palate had a true midline split. Repair of palatal injuries from blunt trauma involved either wire and splinting (eight patients) or miniplate fixation without a splint (11 patients). There were fewer complications in those treated by the miniplate fixation technique (two of 11), as compared to those treated with wire and splinting (four of eight). In miniplate fixation of midfacial fractures, attention is focused on the reconstruction of the supporting buttresses of the midface, both vertical and horizontal. In regard to the palate, this requires union of the inferior horizontal buttress. Gunshot wounds accounted for 21% of the patients and resulted in large bony disruption of the palate, which required free soft tissue and bone grafts for repair of the inferior horizontal buttress. PMID- 8414571 TI - Facial sensibility in patients with unilateral facial nerve paresis. AB - This study evaluated facial sensibility in 29 patients with unilateral lower motoneuron facial nerve paresis using standard clinical tests of sensory evaluation used at other anatomic sites, most commonly the hand. Vibratory and cutaneous pressure thresholds and moving and static two-point discrimination were measured. Statistically significant differences were found between the affected and unaffected sides of the face, with vibration threshold, cutaneous pressure threshold, and static two-point discrimination being greater on the affected side. Vibration thresholds and two-point discrimination (moving and static) progressively decreased, moving down the face from the forehead to the cheek, chin, and then the lip. Sensibility thresholds are altered in patients with unilateral lower motor neuron facial nerve paresis. These findings document a relationship between sensory disturbance and lower motoneuron facial nerve paresis. The potential functional significance of this relationship has clinical significance for patients undergoing rehabilitation training. PMID- 8414572 TI - Temporal bone pathology in scuba diving deaths. AB - Scuba diving has long been associated with otologic injuries; however, little is known about temporal bone pathology in diving-related deaths. We examined 18 temporal bones from 11 divers who died, primarily from complications of rapid ascent. Bleeding into the middle ear and mastoid air cells was nearly universal. Inner ear damage included hemorrhage around Reissner's membrane and the round window membrane and rupture of the utricle and saccule. Most of the observed inner ear damage was not surgically treatable. PMID- 8414573 TI - Formation of mucosal polyps in the nasal and maxillary sinus cavities by infection. AB - Unilateral maxillary sinusitis was experimentally induced in New Zealand White rabbits with Streptococcus pneumoniae serotype 3, Bacteroides fragilis NCTC 9343, and Staphylococcus aureus V8. In another group of rabbits, sinusitis was induced by blocking of the sinus ostium only. Bacteriologic and light microscopic analysis was performed after 5 days to 1 month. Granulation-like polyps developed after deep mucosal inflammatory trauma initiating fibroblast proliferation, angiogenesis, and epithelial migration to cover the polyp. In regions of a more superficial trauma-characterized by epithelial desquamation and fibroblast growth proliferation and differentiation of basal cells resulted in the formation of microcavities dissecting off edematous polyps. Polyps could be found in all sinusitis groups, irrespective of inducing agent. The cellular events of polyp formation appear to be the result of a continuous inflammatory reaction and are not directly related to the presence of a certain microorganism. Instead, the potential of any microorganism to induce a deep mucosal trauma or epithelial desquamation seems essential for its ability to initiate polyp formation. PMID- 8414574 TI - Aneurysm of the thoracic duct presenting as an asymptomatic left supraclavicular neck mass. PMID- 8414575 TI - Nasopharyngeal gliomas: diagnostic and treatment considerations. AB - Nasopharyngeal masses in newborns may produce life-threatening airway obstruction. Distinction between encephaloceles and gliomas is important for surgical planning, but cannot always be accurately assessed on the basis of high resolution computed tomography or magnetic resonance imaging. Preoperative or intraoperative nuclide imaging or contrast injection studies may assist with selection of appropriate surgical approach. PMID- 8414576 TI - Metastatic soft tissue calcification presenting as a tongue mass. PMID- 8414577 TI - Multiple giant pilomatrix carcinomas of the head and neck. PMID- 8414578 TI - Inflammatory pseudotumor of the larynx: comparison with orbital inflammatory pseudotumor with clinical implications. PMID- 8414579 TI - Extruded nasal implant. PMID- 8414580 TI - Tinnitus aureum as an effect of increased tension in the lateral pterygoid muscle. PMID- 8414581 TI - Creation of a knowledge base in otolaryngology-head and neck surgery. PMID- 8414582 TI - Lassa fever and sudden hearing loss. PMID- 8414583 TI - Ventilation tubes and prophylactic antibiotic eardrops. PMID- 8414584 TI - An opinion on perilymph fistulas. PMID- 8414585 TI - IgE-mediated hypersensitivity reactions. AB - The immunologic mechanisms responsible for allergic reactions are well studied and are among the best defined of any immunopathologic disorder. New knowledge is expanding rapidly, and the type I Gell and Coombs immediate hypersensitivity reaction, which is the basis of the IgE-mediated allergic disease, is more accessible to accurate diagnosis and specific therapy than ever before. The mechanisms of the immediate, late-phase, and dual reactions are discussed. The role of the cyclic nucleotide system, nonimmune mast cell degranulation, and participation of cytokines in atopic allergy are investigated. The principles and pitfalls of specific immunotherapy are reviewed briefly. PMID- 8414586 TI - Immunologic changes after nasal provocation with allergen. AB - The high prevalence of allergic rhinitis requires all physicians and surgeons to have a fundamental understanding of this disease. Fortunately, patients with allergic rhinitis can be diagnosed accurately and prescribed effective treatment for symptoms. In the future, as the natural history of allergic rhinitis is appreciated and the pathophysiology better understood, additional points for therapeutic intervention will emerge. Research efforts should focus not solely on acquiring additional symptomatic therapies but also on altering the natural course of the illness. PMID- 8414587 TI - Introduction to in vivo allergy testing. AB - Available in vivo skin tests for assessment of hyperreactivity are essentially similar in principle, in that they rely on the reactivity and sensitivity of dermal mast cells sensitized with specific immunoglobulin E to reflect allergic sensitivity. In this article the immunology of skin testing is examined and the various factors that may influence reactivity are noted. In addition, the respective benefits and drawbacks of such familiar epicutaneous skin tests as the scratch and prick tests and intradermal (or intracutaneous) methods, including skin end point titration, are explored. Efforts to standardize testing technology are also discussed. It is concluded that each method of in vivo skin testing is associated with both advantages and disadvantages and that the most important factors are sensitivity and reproducibility. PMID- 8414588 TI - A comparative study of the diagnostic characteristics of the modified RAST and Pharmacia CAP System. AB - New in vitro technology for the detection of allergen-specific IgE antibody continues to be developed. The manufacturer of one such system, the Pharmacia CAP System, claims that it has test qualities similar to those of the modified RAST. To evaluate this assertion we compared the results obtained with 12 allergens by both procedures on six negative control sera and four sera obtained from atopic patients; a total of 120 tests were performed. The data suggest that attempts to match the sensitivity of the modified RAST are accompanied by a significant loss in test specificity in the Pharmacia CAP System. PMID- 8414589 TI - Chronic sinusitis, steroid-dependent asthma, and IgG subclass and selective antibody deficiencies. AB - In the past 20 years considerable progress has been made in understanding the ways in which subtle immunologic defects can adversely affect health. Immunoglobulin G subclass deficiencies have been identified and are related to an increased susceptibility to respiratory tract infections in certain patients. To assess the immunocompetence of such patients, the quantity and quality of their antibody response must be evaluated. Immunologic evaluation is best performed by measuring selective antibodies before and after a specific challenge. Patients with mild immunodeficiency may benefit from prophylactic antibiotic therapy; those with profound immunodeficiency require antibody replacement therapy. PMID- 8414591 TI - Fighting degradation of medical profession. PMID- 8414590 TI - The role of IgE-mediated hypersensitivity in the development of otitis media with effusion: a review. AB - Otitis media with effusion is the most common cause of hearing loss in children today. This report examines the role of immunoglobulin E-mediated hypersensitivity in the development of otitis media with effusion. It is emphasized that immunoglobulin E-mediated hypersensitivity, or allergy, represents only one variable in a very complex disease entity. Bacterial infection, viral infection, and mucociliary clearance are important variables that must be considered and may be effected by the allergic response. On the basis of the world literature and laboratory investigations at the Children's Hospital of Buffalo, it is concluded that otitis media is associated with allergy in 35% to 40% of cases. Furthermore, the middle ear mucosa itself is rarely a target organ for allergy. Release of biologic mediators of inflammation from basophils and mast cells occurs in the nasal mucosa and nasopharyngeal mucosa. These mediators most likely produce eustachian tube edema and inflammation. Over a long period this chronic inflammatory response, along with viral or bacterial infection, produces middle ear effusion. PMID- 8414592 TI - Not all doctors fit the charges. PMID- 8414593 TI - Pennsylvania takes lead in drug education venture. AB - The Omnibus Budget Reconciliation Act (OBRA) of 1990 required all states to implement drug utilization review programs for outpatient medications covered by Medical Assistance. The State Society is taking a lead role in its response to the retrospective portion of the program. This article explains that role and overviews how OBRA requirements will affect Pennsylvania physicians. PMID- 8414594 TI - Direct-to-the-consumer advertising of drug products. AB - Direct-to-the consumer advertising of prescription drug products is increasingly scrutinized by the federal Food and Drug Administration, spurred in part by growing concern about the potential for abuse posed by such advertising in the absence of effective regulation. Drug industry officials claim they are providing the public with valuable information, but many physicians and health care professionals are distressed by the new trend. PMID- 8414595 TI - A case for tort reform. AB - The current national debate on health care focuses on two issues: access and cost. While Pennsylvania Medical Society positions on health system reform and managed care--overviewed in this publication in January and June 1993 respectively--have dealt with both issues, this article analyzes one specific aspect of the cost issue: the need for tort reform. PMID- 8414597 TI - Caring for the dying patient. Interview by Juli C. McGreevy. PMID- 8414596 TI - Considering AIDS in the elderly. AB - While physicians have been inundated with information about AIDS, they may overlook the possibility of the disease in the elderly, where occurrence is not common. But, as the author points out in the following case study, elderly patients can have the same risk factors for AIDS as younger patients. PMID- 8414598 TI - How are we doing? AB - Following a recent review of the efficiency and direction of this publication, the Society's board directed that a summary of an evaluation completed by the Publication Committee be made available to the entire membership. For new members, this is an explanation of this publication's purpose and form; for all members, this is an open accounting and request for comments. PMID- 8414599 TI - HIV and AIDS in rural Pennsylvania. PMID- 8414600 TI - Understanding the impaired medical student. AB - Medical students encounter some of the stressors of their chosen profession even before receiving their medical degree, and unfortunately they sometimes turn to alcohol and drugs as an escape. This article outlines the stressors of medical school and their impact on students, and offers resources to avoid becoming a victim of impairment. PMID- 8414601 TI - Recognizing, avoiding physician burn-out. PMID- 8414602 TI - A nostalgia trip: has PRO involvement been "worth it"? PMID- 8414603 TI - Vigilance brings victories on economic issues. PMID- 8414605 TI - National Medicare fraud alert issued. PMID- 8414604 TI - Medicare to deny claims from non-CLIA labs. PMID- 8414606 TI - Pondering dangerous waters. PMID- 8414607 TI - Considering the basics of system reform. PMID- 8414608 TI - Alternative advocated to finance reform. PMID- 8414610 TI - What your Society likes in the Governor's plan. PMID- 8414609 TI - Caring for the health and culture of migrant workers. PMID- 8414611 TI - An analysis of cesarean deliveries among Pennsylvania Blue Shield subscribers. AB - In recent years, increased attention has focused on the relatively high rate of Cesarean deliveries in the U.S. Though the medical necessity and indications for Cesarean deliveries have been heavily debated, there still remains considerable disagreement among professionals about which Cesarean deliveries are inappropriate. To contribute to an understanding of these issues, Pennsylvania Blue Shield prepared a profile of Cesarean deliveries in Pennsylvania relative to Blue Shield's claim experience. The results of the study are presented in this article. PMID- 8414612 TI - Screening lab studies in nursing homes: are they useful? PMID- 8414613 TI - A new PRO direction. PMID- 8414614 TI - Violence prevention in Philadelphia. Interview by Peter B. Kang. PMID- 8414615 TI - Health care needs of State's American Indians addressed. PMID- 8414616 TI - New doctors face stagnant market. The Health Care Group. PMID- 8414617 TI - From one special interest to another. PMID- 8414618 TI - [Nutritional and fluid requirements of the child involved in sports]. AB - Between generally accepted recommendations and individually ascertained values for need of energy, nutrients and water, which are evaluated more exactly only by laboratories of nutritional physiology and of sportsmedicine, wide variations are possible. For practical purpose regular controls of physical development, competitiveness, regular weighing, measuring and testing alone, give the possibility to assess, whether food-intake meets quantitatively the need. Qualitatively deficient nutrient-intake is only shown after some time elapsed by reduced competitiveness or reduced nutritional recommendations are to performed, which are summarized in the ten rules of prudent nutrition (DGE). In competitive sports, basic for optimal, sporty nutrition are controls of physical development and of nutritional intake (nutritional anamnesis). To avoid nutritional deficiencies and their consequences in children doing intensive sport, following minimal demands should be performed: 1. Children, regularly highly trained are classified to "children at risk" and are to be looked after by physician with a degree in sportsmedicine, familiar with the development of children (pediatrician). 2. Physical development of children performing competitive sports has to be investigated and documented in two-monthly periods (training-protocols, growth-charts) minimal. 3. Minimum twice a year, by a nutritional well qualified person (physician in sports, nutritional scientist, dietitian) a nutritional advisory has to be done to the children together with parents and trainer. 4. If physical development or competitiveness will be disturbed, the individual need of energy and nutrients has to be ascertained, basic for an following nutritional advisory. 5. Assumption of energy-need for children performing competitive sports has to be done individually. Basic for assumption is the "biological age".(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414619 TI - [The Nutrition Committee of the Austrian Society of Pediatrics. Antigen-reduced infant nutrition]. AB - In infants at risk of developing allergies exclusive breast feeding during the first 4-6 months and introduction of solids after four months of age reduces the incidence of atopic syndromes at least up to 36 months of age. The protective effects of two antigen reduced infant formulae (marketed as "Hypoallergenic" in Austria) against atopy in infants at risk have been confirmed in longitudinal studies. The incidence of allergic symptoms was significantly reduced during the first 18-36 months if introduction of solids was delayed until 4-6 months of age. Antigen-reduced infant formulae and no solids should be given to infants at risk who are not exclusively breastfed during the first 4-6 months. PMID- 8414620 TI - [Publications from Austrian pediatric institutions--summary and analysis 1989 to 1991]. AB - Between 1989 and 1991 16 Austrian pediatric institutions published 232 scientific papers with a total impact-factor of 200,406 in internationally recognized journals. Seventy-three percent of them came from only three institutions, the University departments of pediatrics in Graz and Vienna and the St. Anna Children's Hospital (77, 63 and 30 publications, respectively). The impact-factor (IF) of these publications ranged from 7.591 to 0.010. The average IF was 0.864 and the median IF 0.430 per paper. Whereas 70 papers (30%) were published in journals with an IF over 1, only one third appeared in pediatric journals. PMID- 8414621 TI - The clinical efficacy of Vertigoheel in the treatment of vertigo of various etiology. AB - In this paper the authors describe the clinical efficacy in treatment of vertigo of various etiology. A group of 31 patients were treated with Vertigoheel medication: 14 patients suffered from vertebrobasilar arterial insufficiency, 8 patients were diagnosed as Meniere's disease, 5 patients complained of vertigo of traumatic origin and 4 patients suffered from neuronitis vestibularis. The authors found regression of clinical symptoms in the majority of cases in the investigated group who were treated with Vertigoheel. PMID- 8414622 TI - Perinatal mortality and morbidity in premature birth. AB - In a study carried out in 100 cases of premature birth in Section "A" of the Department of Gynecology and Obstetrics of the University of Turin, the authors report a perinatal mortality rate of 5.4%. Perinatal morbidity was found to be very high (68.2%). These findings--which are perfectly in line with those reported in the literature--explain why premature birth is one of the most important topics of research in modern obstetrics. PMID- 8414623 TI - Evaluation of chemical parameters in breast cyst. AB - During our studies on breast diseases, the following biochemical values were measured: chlorine, sodium, potassium and albumin. We examined 153 syringe biopsies from women aged between 34 and 55 years, and an analysis of the above mentioned parameters enabled us to classify the breast cyst fluid into 2 groups: group I was definitely benign, while group II was calculated as having a risk factor two to four times greater than the control group in developing breast cancer. PMID- 8414624 TI - Is there really any significant link between a high fat diet and breast cancer? AB - The possibility of a connection between a diet high in saturated fatty acids and breast cancer is examined and is found to lack conclusive proof as yet. Prevention by means of early clinical and instrumental examinations appears to offer a greater chance of success. PMID- 8414625 TI - Proton-therapy, present status. AB - At the moment, proton-therapy is the most advanced radiotherapeutic technique in cancer treatment. The use of the high energy proton beam (from 70 MeV to 200 MeV) lets a Bragg's peak be moved to different depths, so allowing personal radiotherapeutic treatment. In recent years, many proton-therapy centers have grown up throughout the world with very satisfactory clinical results, first of all in eye melanoma treatment. The future expectations are very promising, even if the very high installation and maintenance expenses of a synchrotron (for proton production) hinder the development of such a method. PMID- 8414626 TI - The artificial endocrine pancreas in clinical practice and research. The present position and future perspectives. AB - The artificial endocrine pancreas (AEP) is a controlled glucose and/or insulin infusion system in which continuously monitored blood sugar values are fed to a computerised analyser that uses predetermined algorithms to establish the doses to be administered. Since its first appearance in clinical practice and diabetological research during the Sixties, the AEP has been modified in various ways to overcome technical problems associated with the gluco-sensor and algorithms so as to make better use of the glucose-insulin feedback mechanism, and hence obtain a closer correspondence to physiological islet cell activity. As a result of these changes, the AEP can be employed in accordance with the physiopathological principles of insulin secretion in a variety of clinical conditions to secure the short-term control of metabolic alterations in the diabetic. Surgery is one field in which the AEP is used to great advantage, since this and its accompanying anaesthetics are the source of stress, which in turn may result in a rapid and sometimes serious postoperative metabolic derangement, including an increased secretion of anti-insulin hormones. The AEP has also been proposed for diabetic pregnancy and for the treatment of subjects in diabetic coma. It has proved useful in the diagnosis and management of hypoglycaemia due to organic hyperinsulinism, in diabetics with renal failure, in the honeymoon period, and in cases of unstable diabetes. The versatility of its application and its underlying physiopathological principles have enabled the AEP to be predominantly employed in research.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414627 TI - Inter/intra-observer variability of carotid and femoral bifurcation intima-media thickness measurements. AB - Arterial wall thickening may be quantitatively assessed by measuring the intima media thickness (IMT) with high resolution ultrasound. Previous studies have shown a good inter/intraobserver variability of IMT measurements in the common carotid. In this study we evaluated the inter/intraobserver variability of IMT measurements in 10 randomly selected asymptomatic subjects (age 55.4 +/- 6). Two carotids and two femorals were studied in each subject. IMT for each patient was the average of five IMT measurements at the artery bifurcation. Three observers repeated the scanning and the measurements twice with no knowledge of the previous readings. The between observer coefficient of variation (CV) was 8.45%; the intraobserver CV (mean of carotids and femorals) varied from 4.4 to 5.1% for the three observers who measured IMT three times. The mean absolute difference between the first and the third measurement was 0.0738 mm. In conclusion IMT measurement variability is mostly due to differences between observers. The intraobserver variability is very small. IMT measurements at the carotid and femoral bifurcations have a low variability and are a good expression of atherosclerosis as they consider early lesions at the bifurcation level which may not be observed in the common carotid. PMID- 8414628 TI - Immunological and clinical surveillance in Recklinghausen's neurofibromatosis (NF1). AB - Neurofibromatosis evolution is described in relation to the factors that may favour its expansion and immune modifications. NF1 monitorization should employ periodic clinical and immune surveillance. Such an approach would allow the application of immunomodulating treatment (e.g. adapted therapy) only when indicated, thereby reducing its duration and potentiating its efficacy. PMID- 8414629 TI - Intraluminal high dose rate brachytherapy combined with external radiotherapy in the treatment of oesophageal cancer. AB - Twenty eight patients with previously untreated oesophageal carcinoma without distant metastases were divided into two groups: Group A consisted of 18 pts. treated with conventional external radiotherapy only. Another group of 10 pts. (Group B) received treatment with external beam irradiation with further high dose rate intraluminal brachytherapy up to a dose of 4-12 Gy delivered in 2-3 sessions of 4 Gy (one session a week). All pts. were evaluated clinically, radiologically and endoscopically every 3 months. At the end of treatment there was a marked difference in relief of dysphagia (39% in Group A vs. 90% in Group B), local control (56.7% in Group A vs. 100% in Group B) and time to progression of dysphagia (20.8 weeks in Group A vs. 67.7 weeks in Group B). No marked difference was observed in overall survival. The complication rate was low in both groups and major complications were observed in pts. treated with external radiotherapy alone (two fistulas). The association of external beam and intraluminal radiotherapy can give a better local control of the disease, improving the quality of life. PMID- 8414630 TI - Transfer in vitro of tubercolin hypersensitivity by sensitised lymphocytes. AB - An account is given of the experimental in vitro transfer of PPD-specific delayed hypersensitivity to peripheral leukocytes pulsed with a very small portion of autologous lymphocytes pre-sensitised with PPD-specific transfer factor (TF). Transfer was assessed by means of the leukocyte migration inhibition test, the lymphocyte locomotion test, and the leukocyte adherence inhibition test. These tests were positivized in all the experiments. It is suggested that these results indicate that TF does not act in itself, but triggers a reaction that expands the effect of hypersensitivity. PMID- 8414631 TI - Two-point discrimination thresholds in spinal cord injured patients with dysesthetic pain. AB - We questioned whether deafferentation following SCI would result in an increase in somatic sensitivity possibly due to cortical reorganization. Dysesthetic pain syndrome (DPS) below the level of a spinal cord injury (SCI) is a common complication. We hypothesized that DPS patients would show increased cortical reorganization because of high levels of sensory stimulation following injury. Sixteen dysesthetic pain SCI patients, 15 SCI patients without pain, and 16 control subjects were examined for two-point discrimination thresholds (2PDT) of the forearm, neck, and spine. The SCI pain group had significantly smaller 2PDTs than either SCI no pain or control groups, particularly over the neck and spine. The SCI pain group had a significant inverse correlation between perceived degree of pain (visual analogue scale) and 2PDT in the spinal skin area. The findings indicate that SCI patients with severe DPS have a higher sensitivity to somatosensory stimuli, particularly in skin areas with projections to primary somatosensory cortex areas adjacent to the deafferentated region. The increase in 2PDT may be due to an increase in the size of the somatosensory cortical areas allotted to the corresponding skin areas. PMID- 8414632 TI - Bacterial biofilm formation on the bladder epithelium of spinal cord injured patients. II. Toxic outcome on cell viability. AB - As a follow up to our first study of 10 spinal cord injured patients, a further 8 patients were investigated over 2 months for biofilm formation on their bladder epithelial cells and for evidence that these uropathogens damage the host bladder. All the patients were found to be colonized with uropathogens, regardless of whether or not they were receiving antibiotics. Using vital staining, it was discovered that there was a significant reduction (33%) in bladder cell viability in the presence of bacterial biofilms compared to controls. This was not associated with cell turnover rates. In vitro tests showed a similar reduction in cell viability when uropathogens were incubated with bladder cells. In addition, white cell counts were significantly elevated in the patients' urine, indicative of an infectious and/or inflamed state. There was no difference between symptomatic and asymptomatic patients in their mean bacterial adhesion counts. Patients were just as likely to be symptomatic as asymptomatic when on antibiotics. In summary, the presence of virulent organisms in the bladder does adversely affect the host, even when the patient has insignificant signs and symptoms of infection, thereby raising concerns over the decision not to treat the patient. Unless specific antibiotics are used which eradicate adherent biofilms from the bladder, the treatment of symptomatic patients will only impact upon the signs and symptoms in some patients, and not alter their susceptibility to reinfection. PMID- 8414633 TI - Ultrasound in the early diagnosis of heterotopic ossification in patients with spinal injuries. AB - Heterotopic ossification (HO) is a potentially disabling complication of spinal injuries and other chronic disorders. It is of unknown aetiology and currently there is no easy or convenient diagnostic method that will allow very early confirmation of the inflammatory changes that precede osteoid and, later, true bone formation. Clinical experience, however, indicates that early treatment with radiotherapy, antiinflammatory agents or diphosphonates is needed to control the progression. This study was undertaken to assess the role of ultrasound (US) in the very early diagnosis of HO in patients with spinal injuries. US was found to be very sensitive in detecting focal soft tissue abnormalities around joints and in the muscles of these patients. If combined with a Doppler study to exclude deep venous thrombosis (DVT), and infection or tumour could be excluded clinically, US was extremely accurate in predicting the presence or absence of early HO changes within hours of the clinical manifestation. In 2 patients it successfully predicted HO in the opposite leg before clinical signs were evident. This study also provided supportive evidence of the theory of microtrauma in the aetiology of HO. As ultrasound is portable, safe, cheap, reproducible and accurate, it is the method of choice in the early diagnosis of HO. It allows early treatment to prevent the formation of osteoid and subsequent bone formation. PMID- 8414634 TI - The association between deep venous thrombosis and heterotopic ossification in patients with acute traumatic spinal cord injury. AB - The medical records of 209 patients with acute traumatic spinal cord injury (SCI) admitted to the SCI rehabilitation unit from 7/1/88 through 12/31/92 were reviewed. Whereas the incidence of heterotopic ossification (HO) and deep venous thrombosis (DVT) in this population were 16.7% and 14.3%, respectively, 36.6% of the individuals with DVT had HO. 31.4% of those with HO developed DVT at some time during their acute or rehabilitation hospitalization. The overall incidence of coexistent DVT and HO was 5.3%. The correlation between the occurrence of HO and DVT in this SCI population reached statistical significance (chi 2 = 9.97; p < 0.005). The results of this study suggest that there exists an association between the occurrence of DVT and HO following traumatic SCI. We hypothesize that venous compression from expanding heterotopic bone can result in lower limb DVT following traumatic SCI. PMID- 8414635 TI - Indomethacin and disodium etidronate for the prevention of recurrence of heterotopic ossification after surgical resection. Two case reports. AB - Two spinal cord injured patients had extensive heterotopic ossifications around their hips surgically removed. The use of disodium etidronate from 2 weeks presurgery to 2-5 months postsurgery and in particular indomethacin 25 mg twice daily, from 2 weeks prior to surgery to 6 weeks postsurgery may have helped to prevent recurrence of the ossifications in the follow up periods of 4 and 5 years respectively. PMID- 8414636 TI - Intrathecal baclofen: does tolerance occur? AB - Concern over the development of tolerance in patients on continuous intrathecal baclofen therapy has arisen as this new form of treatment for spasticity has gained wider use. We have studied time-dose relationships in 18 spinal cord injured patients who have undergone intrathecal baclofen infusion pump implantation since February 1988 in our facility. Our data show that there was a significant increase in baclofen dosage needed to control spasticity during the first 12 months post implantation. After 12 months, however, no significant changes in dosage requirement was detected. In addition, there was no significant difference between completely and incompletely spinal cord injured patients with regard to both the initial dose and the tolerance trend. PMID- 8414637 TI - Red Cross spinal injury project in Bucharest, Romania. AB - Inadequate medical facilities with lack of specialised drugs and other essential medical items, scientific isolation of medical staff, unavailability of orthoses and wheelchairs: such were a few of the problems identified in Bucharest in July 1991. The aim of the federation at the start of the project was to launch an action in many directions using two basic principles: using local means including human resources, and improving the existing facilities instead of erecting new buildings. Training of the staff goes side by side with the practice of total initial care. The family is taught to position and turn patients, practice chest physiotherapy and has become a major psychological and emotional support. An orthopaedic workshop has been set up to provide orthoses and to supervise the construction of wheelchairs in a local factory. The social situation of patients is assessed to initiate social integration in the community. Home visits are organised to offer advice for adaptations. The implementation of these actions is difficult in a country which has to solve many economic problems. PMID- 8414638 TI - The causes, sequelae and attempts at prevention of cervical spine injuries in Poland. AB - There are certain differences between the causes of spinal injuries in Poland and in the developed countries of the West. The most common causes of injuries to the cervical spine in Poland are: falls from a horse-cart, diving into shallow water and automobile accidents. This paper presents an analysis of the causes of cervical spine injuries in a series of 1937 patients treated at the Spinal Cord Injury Department of the Metropolitan Rehabilitation Center in Konstancin, Poland, in the years between 1965 and 1991, admitted to hospital within the first hours or days after spinal injury. The relations between the mechanism of injury and the cause of injury, as well as the neurological sequelae of the injuries, are analysed. PMID- 8414639 TI - Electrical stimulation-assisted rowing exercise in spinal cord injured people. A pilot study. AB - Recently a FES (functional electrical stimulation)-assisted rowing machine was developed to enhance cardiovascular training in people with spinal cord injuries. The machine was assessed in terms of its efficacy as a training tool. Six patients who were quadriplegic (C6-T1) and 2 who were paraplegic (T3-6) completed a series of three tests in succession: (1) leg stimulation only (quadriceps and hamstring groups)--'Stim', (2) arm row only--'Row' and (3) simultaneous row and stimulation--'R & S'. Measurements recorded included oxygen uptake (VO2), minute ventilation (Ve), respiratory exchange ratio (RER), heart rate (HR) and blood pressure (BP). In addition, 6 out of the 8 subjects took part in a qualitative assessment comprising a guided interview exploring the subject's perception of the machine and test. Significant increases in VO2 were demonstrated between the three tests with R & S producing mean steady-state values of 16.34 nm (+/- 0.74) ml/kg/min (83% of maximum). These values represented a 12% increase over Row alone. Of interest was the qualitative assessment which revealed that subjects perceived R & S to be easier than Row despite the higher levels of VO2 observed. The results suggest that the rowing machine represents a potentially valuable hybrid training device that may significantly reduce risk factors for cardiovascular disease and improve the quality of life of people with SCI. PMID- 8414640 TI - Complete dislocation with burst fracture of the lower cervical spine. Case report. AB - A patient with a complete fracture dislocation of the C6 vertebra without any neurological deficit is presented. The absence of neurological changes in an area of the spinal canal which is almost completely filled with the spinal cord is remarkable. The mechanism of injury in this patient is believed to be hyperextension associated with significant axial loading. Hyperextension first ruptured the anterior and the posterior longitudinal ligaments and then injured the pedicles of C6. The associated axial loading resulted in a 'bursting' of the posterior elements, thereby providing room for the spinal cord to move dorsally with the avoidance of a major neurological deficit. PMID- 8414641 TI - Localization of the A1.12/9 antigen family to the neurones, putative sensory receptors and tegument of Schistosoma mansoni. AB - The A1.12/9 antigen family were localized to the periphery of vesicle-like structures in neurones and in the putative sensory receptors of cercariae by immuno-electron microscopy. After transformation into schistosomula, the antigens were rapidly lost until, after 18 h, no immunoreactivity could be detected. Expression resumed at approximately 36 h post-transformation and had returned to high levels by 4 days post-infection. This level was maintained through to the adult worm stage. In 4 day lung schistosomula and in 14 day and 21 day liver schistosomula immunolabelling was observed within the matrix of the tegument itself and also in association with the tegumental membrane and vesicles within the tegument. These properties suggest that the A1.12/9 antigens may be involved in neuropeptide processing pathways. In particular, this family may represent the schistosome homologue of the granin family with which they share common properties and some sequence homology. PMID- 8414642 TI - Suppression of Schistosoma bovis egg production in cattle by vaccination with either glutathione S-transferase or keyhole limpet haemocyanin. AB - Two of the antigens which have shown vaccine potential in animal experiments against Schistosoma mansoni are glutathione-S-transferase (GST) and GP38, protective epitopes of which are shared with keyhole limpet haemocyanin (KLH). We therefore tested S. bovis GST and KLH for vaccine efficacy against S. bovis in the natural Zebu cattle host. In a preliminary experiment three vaccinations with a total of 1.39 mg of native GSTs of S. bovis induced specific antibody at the time of challenge as detected by Western blotting and ELISA and mean faecal egg counts between weeks 6-10 post-challenge were reduced by 56.4 to 82.5% compared to non-vaccinated controls. Mean adult worm recoveries and tissue egg densities in large intestine and liver samples were also reduced in the vaccinated group, but these differences were not statistically significant. In a subsequent experiment one group of calves was vaccinated with a similar schedule to that used above; a second group of calves was given only two injections of GST (total 0.48 mg protein); a third group of calves was vaccinated twice with a total of 2.0 mg KLH in PBS. All three vaccination schedules induced specific antibody. Both GST vaccination schedules induced significant reductions in faecal egg counts compared to non-vaccinated controls and in this experiment tissue egg densities were also significantly reduced. A striking finding, however, was that adult worm counts were not reduced by vaccination. An essentially similar outcome resulted from KLH vaccination, since there were significant reductions in faecal and tissue egg counts in the absence of a reduction in adult worm numbers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414643 TI - Immunological relationships during primary infection with Heligmosomoides polygyrus (Nematospiroides dubius): parasite specific IgG1 antibody responses and primary response phenotype. AB - IgG1 antibody responses to Heligmosomoides polygyrus were measured in eight mouse strains supporting acute (< 8 weeks, SJL, SWR), intermediate (10-20 weeks, NIH, BALB/c) or chronic (> 25 weeks, C57BL/0, CBA, C3H, AKR) primary infections. Mice supporting acute or intermediate infections produced more intense antibody responses and total serum IgG1 concentrations were higher than in mice tolerating chronic infections. Positive correlations across mouse strains between the intensity of the antibody response and the percentage loss of worms in weeks 6 and 10 were established. No correlation was found between the response within mouse strains and loss of worms by individual mice. Heavy infections gave marginally higher antibody titres than low intensity infections, but few significant differences were detected and it was concluded that infection intensity did not markedly influence the magnitude of the antibody response. Male and female mice responded similarly despite the earlier loss of worms from females. No association was found between the primary response phenotype and recognition of particular antigens in Western blot analysis, nor did intensity of infection or host gender affect recognition. The possibility that immunomodulatory properties of adult worms may have had a differential influence on ability of strains of contrasting response phenotype to mount IgG1 responses was discussed. PMID- 8414644 TI - Immunological relationships during primary infection with Heligmosomoides polygyrus (Nematospiroides dubius): downregulation of specific cytokine secretion (IL-9 and IL-10) correlates with poor mastocytosis and chronic survival of adult worms. AB - Mice were infected either with Trichinella spiralis (day 0), Heligmosomoides polygyrus (day-14) or concurrently with both species and were killed in groups, together with naive control mice, on 2 occasions (day 8 and 15 post infection with T. spiralis, corresponding to days 22 and 29 p.i. with H. polygyrus). The expulsion of T. spiralis was slowed significantly in concurrently infected mice and this was associated with a reduced mastocytosis and lower serum mucosal mast cell protease levels. Mesenteric lymph node (MLN) lymphocytes from all three experimental groups secreted IL-3 and IL-4 in copious amounts when stimulated in vitro by Concanavalin A (Con-A), but the secretion of high levels of IL-9 and IL 10 was essentially confined to mice infected with T. spiralis alone. It is suggested that adult H. polygyrus selectively modulate cytokine secretion by Th2 cells within the MLN during infection and that this is brought about as a direct consequence of the mechanism employed by H. polygyrus to depress mucosal inflammatory responses in order to facilitate its own survival. PMID- 8414645 TI - The effect of irradiation on the third stage larvae of Brugia pahangi. AB - The effect of irradiation on the third stage larvae of the filarial nematode Brugia pahangi was investigated. Labelling with 35S methionine of control or irradiated L3, post-infective L3 or L4 revealed no consistent alterations in the pattern of proteins synthesized. The only significant difference observed was in 125I labelling, where the specific activity of labelling of soluble cuticular proteins was lower in irradiated than in control parasites. This difference may be related to the reduced size of irradiated parasites rather than to a specific effect of irradiation on the expression of cuticular proteins. Irradiated parasites recovered on day 14 post-infection were significantly shorter than control parasites. Irradiation also appeared to have a lethal effect on male parasites, as no recognizable males were recovered from animals given irradiated L3, nor were microfilariae ever observed in these animals. The mechanisms by which irradiation may enhance the immunogenicity of L3 of filarial nematodes are discussed. PMID- 8414646 TI - [Autogeny in bloodsucking biting midges of the genus Culicoides (Diptera: Ceratopogonidae)]. AB - The phenomenon of autogenity has been examined by means of different criteria both in nature and in laboratory in 25 blood-sucking species of the genus Culicoides, which were collected in Russia, Uzbekistan, Azerbaijan, Armenia, Georgia and Kazakhstan during 1979-1990 years. An autogenetic maturation of eggs has been observed in 6 species, and the absence of this phenomenon has been observed in 19 species. It is suggested, that autogenity or non-autogenity is a species characteristic of blood-sucking midges. The processus of autogenetic vitellogenesis takes the time from later pupa (more than 2 days of this stage) to earlier imago (lesser than 2-3 days of this stage). PMID- 8414647 TI - [The influence of reproductive system function in female mosquitoes on their capacity for infection with the causative agent of malaria]. AB - The study of the vector-agent model (Anopheles sacharovi-Plasmodium gallinaceum) has shown that the development of ovaries of the vector stimulates its receptivity to the malaria agent. In the case of braking of follicle development the mosquitos were nearly not infected. The capability of mosquitos to be infected without previous carbohydrate feed is confirmed. The influences of the age of mosquitos and the quantity of blood soaked up on the egg development and on the receptivity to malaria agent are demonstrated. PMID- 8414648 TI - [2 new species of mites in the genus Neotrombicula (Trombiculidae) from the western Caucasus]. AB - Two new species of the genus Neotrombicula Hirst, 1925 are described from Western Caucasus. Neotrombicula alexandrae sp. n. is similar to N autumnalis (Shaw, 1790) but differs from it by several measurable characters and by sensillae barbed only in distal half. N. abagoensis sp. n. slightly resembles N. autumnalis and differs from it by significantly greater value of the majority of standard measurements, NDV and by more thick barbs of D. Specimens of new species were collected on rodents near the towns Maikop, Anapa, and in the area of the Caucasian reserve. All holotypes and paratypes of new species are deposited in the Zoological Institute, Russian Academy of Sciences (Saint-Petersburg). PMID- 8414649 TI - [The virulence, metacyclogenesis and respiratory enzymes of Leishmania isolated in culture from laboratory animals]. AB - It has been shown that an increase of virulence of Leishmania major, L. tropica, L. braziliensis as a result of passing through animals and its decrease during the cultivation are accompanied by certain changes of biochemical characteristics of these promastigotes. In the former case the activity of NADP-H-diaphorase and peroxidase of promastigotes and their ability to be transformed into final (invasional) metacyclic forms increase and in the latter case these characteristics decrease. The level and duration of virulence in culture depend not only on absolute value of the above-mentioned characteristics but also on the graduality of their change. Metacyclogenesis and activity of oxidative enzymes are suggested to be the correlates of virulence of various Leishmania species. PMID- 8414650 TI - [One more biological characteristic of coccidia in the genus Cryptosporidium (Sporozoa: Apicocomplexa)]. AB - Suckling rats were used as a model host for our previous EM studies of the endogenous development of Cryptosporidium parvum isolated from spontaneously infected calves (Beier e. a., 1990; Beier, Sidorenko, 1990). In the course of repeated infections it was noticed that the oocysts discharged by the recipient host (rat) were obviously larger than those in the donor's (calf's) isolates. Keeping in mind the presumable taxonomic significance of coccidian oocysts as a constant and rather reliable tool for species discrimination we made a comparative quantitative and statistical analysis of the mean values of oocysts of C. parvum, originating from several sources (table 2): a random human isolate (N 1), several random isolates from spontaneously infected calves (N 2-5), isolates from calves (N 6-8) used as the infecting material for suckling rats, and fecal samples from the experimentally infected rats (N 9-15). The results obtained have shown that the oocysts discharged by rats (N 9-15) were larger that those of calf origin (N 6-8), with the differences being statistically significant with 95% confidence. Besides, within the same host (rat) at least two oocyst groups were distinguished (N 9-11 and 12-15, resp.) whose differences in mean values also appeared statistically significant. The larger oocysts displayed differences in morphology bearing distinct walls which were never observed either in the donor isolates or in the smaller population of the recipient oocysts. The established differences in oocyst dimensions lay presumably within the frames of the normal reaction of C. parvum and other Cryptosporidium species, due to biological peculiarities of unusual life cycles of these unique coccidia: their homoxenous (i.e. confined to one host body only) development is combined with polyxeny (i.e. a wide host specificity that involves the number of host species representing different environmental conditions). The parasite's ability to change its functional morphology and size may appear some kind of preadaption to the number of varying conditions met by these polyxenous coccidia. This and other relevant assumptions are discussed in the paper. PMID- 8414651 TI - [Lactate dehydrogenase activity and changes in its isoenzyme spectrum in the liver of the bream Abramis brama parasitized by pleurocercoids of the cestode Digramma interrupta]. AB - Changes of lactate-dehydrogenase (KF 1.1.1.27) activity have been studied in the liver of the bream been infected with pleurocercoids of the cestode Digramma interrupta. It is showed, that an increase of enzyme activity depends upon a slowly migrating from LDH-6. Problems of interaction at the level of substrate within a system parasite-host is considered. PMID- 8414652 TI - [The chromosome sets of trematodes]. AB - This report provides the data on diploid and haploid chromosome number and structure of karyotypes of 230 species of trematodes. Also, it presents here original author's materials on peculiarities of chromosome sets of 88 trematode species, most of which have been studied for the first time. PMID- 8414653 TI - [A new design of canopied counting trap for estimating the number of bloodsucking Diptera]. AB - A modified trap for quantitative estimations of bloodsucking insects is described. It is shown, that this trap has a number of advantages. Technical description, recommendations on constructing and exploitation are provided. PMID- 8414654 TI - Surgical techniques for arthroscopic meniscal repair. AB - Meniscus repair surgery should involve: (1) concomitant anterior cruciate ligament (ACL) reconstruction in ACL-deficient knees, (2) rasping of the perimeniscal synovium and both tear surfaces, (3) the use of a posterior incision and popliteal retractor, (4) closely spaced, vertically placed suture repair with good coaptation of the tear surfaces, (5) the implantation of an exogenous fibrin clot in the defect, and (6) a well-supervised rehabilitation program. This article reviews the different surgical techniques for arthroscopic meniscal repair. PMID- 8414655 TI - Selection criteria for knee arthroscopy in the osteoarthritic patient. AB - In the current literature concerning arthroscopy in the osteoarthritic patient, there are few well-controlled studies with long-term follow-up that establish selection criteria. The effectiveness of abrasion arthroplasty has not been proved in prospective studies. Prospective factors that are associated with a better outcome include normal limb alignment, a history of mechanical symptoms, minimal roentgenographic degeneration, and a short duration of symptoms. Variables associated with poor outcomes include varus or valgus malalignment, loading symptoms, severe roentgenographic degeneration, previous surgeries, and chronic symptoms. Advanced age, per se, is not a contraindication to arthroscopy. PMID- 8414656 TI - Treating gunshot femoral shaft fractures with immediate reamed intramedullary nailing. AB - Reamed intramedullary nailing was used within 36 hours of patient admission to treat 32 consecutive femoral shaft fractures caused by low-velocity gunshot wounds. The femoral shaft fracture was classified according to the AO system, and comminution was graded according to the classification of Winquest and Hansen. Patients were followed for an average of 14.7 months (range, 6 to 36 months), and the average time to union was 18.6 weeks. The average hospital stay was 7 days, which is approximately one half the average stay reported in the literature for treating this kind of injury with delayed nailing. The shorter hospital stay represents potential savings of up to $9,000 per patient. Immediate intramedullary rodding is a safe, effective, and economic option for the treatment of a femoral shaft fracture caused by a low-velocity gunshot wound. However, a lack of compliance with instructions concerning weight-bearing in this patient population needs to be taken into account when planning postoperative care. PMID- 8414657 TI - Sacroiliac joint pyarthrosis. AB - Sacroiliac (SI) joint infection is rare, and symptoms are idiosyncratic and often confusing. This paper reviews six cases of SI joint infection with regard to their clinical manifestations, diagnostic imaging, and treatment. The six patients were seen over a 15-year period and had a mean age of 16.2 years and a mean follow-up of 44.2 months. The most common presenting symptom was fever (71.4%). The most common physical findings were elevated temperature and limited ipsilateral hip motion. In most cases the white blood cell count and erythrocyte sedimentation rate were elevated, and 71% of the cases had positive blood cultures. The most specific imaging study was the technetium bone scan. All patients were treated with 3 to 6 weeks of appropriate antibiotics and were asymptomatic and had a normal physical examination on follow-up. Routine anteroposterior roentgenograms of the pelvis demonstrated sclerosis of the affected SI joint in one third of the cases. There appeared to be no long-term sequelae in this group of patients. PMID- 8414658 TI - Is pinning in situ a safe procedure for slipped capital femoral epiphysis? AB - This report reviews the results of 37 patients with 46 involved hips treated with pinning for slipped capital femoral epiphysis. Particular attention was given to incidence of complications, including avascular necrosis, chondrolysis, and penetration of the joint. Of 46 hips with 1-year follow-up, there were no cases of chondrolysis. Of 23 hips with 2-year follow-up, there was only 1 case of avascular necrosis. Pin penetration, recognized at the time of surgery, produced no significant sequelae. Pinning can produce satisfactory results without a significant incidence of major complication. PMID- 8414659 TI - Methohexital for orthopaedic procedures in the emergency department. AB - The past two decades have seen more and more orthopaedic procedures performed in the emergency department. Methohexital would seem to be a useful adjunct drug for the performance of these procedures because of its well-known attributes (eg, rapid induction and recovery, brief duration, and minimal hemodynamic changes). A search of the literature revealed no previous studies on the use of methohexital in the emergency department. Therefore, the authors undertook a 1-year prospective study of all patients in their emergency department who received methohexital for orthopaedic procedures. The study's hypothesis was that methohexital is a safe drug for use in orthopaedic procedures in the emergency department. Additionally, the authors sought to determine the drug's indications for use, patterns of usage, and effects on the respiratory and cardiovascular systems. The data presented here are a subset of data previously presented and published. PMID- 8414660 TI - Old displaced fracture of the scaphoid. An unusual cause of carpal tunnel syndrome. AB - The case of a 37-year-old man with carpal tunnel syndrome and a palpable mass in the proximal wrist is reported. Roentgenograms revealed an old middle-third scaphoid fracture with the proximal pole displaced into the proximal carpal tunnel. Excision of this mass led to a complete resolution of the patient's carpal tunnel problem. PMID- 8414661 TI - Management of a multiply injured Jehovah's Witness with severe acute anemia. AB - The refusal of blood products by Jehovah's Witnesses creates ethical and medicolegal dilemmas for the treating physician. Appropriate management involves some understanding of the beliefs of the Jehovah's Witnesses and knowledge of a variety of techniques to minimize blood loss. This case report describes the treatment of a Jehovah's Witness with severe anemia and multiple skeletal injuries. The need to keep blood loss to a minimum influenced the management of this patient. PMID- 8414662 TI - Luxatio erecta: persistent displacement of the greater tuberosity after reduction. AB - The case of a 36-year-old man who sustained a luxatio erecta dislocation of his shoulder is reported. He had persistent displacement of a fracture of the greater tuberosity after closed reduction of the shoulder. The two-part fracture required operative treatment. This is the first occurrence of this complication reported in the literature. PMID- 8414663 TI - A simple way to make traction pins safer. AB - Prevention of inadvertent injuries to patients and hospital staff becomes more important than ever in today's hospital environment. The sharp ends of traction pins, a previous hazard, can be made safe using materials easily found throughout the hospital. Our method uses items that are easily replaced and easy to apply. PMID- 8414664 TI - A 77-year-old man with a gradually enlarging mass of the left ring finger. AB - The following case is presented to illustrate the roentgenographic and clinical findings of a condition of interest to the orthopaedic surgeon. Initial history, physical findings, and roentgenographic examinations are found on the first page. The final clinical and roentgenographic differential diagnoses are presented on the following pages. PMID- 8414665 TI - A skin and soft-tissue protector for intramedullary reaming. AB - A 60-mL plastic syringe can be modified to make a reliable and inexpensive skin and soft-tissue protector for use during intramedullary reaming of the femur. This device is effective and simple to use. It frees the surgeon's hands during reaming and does not require any attachment to the drapes to prevent the device from falling onto the operating room floor. PMID- 8414666 TI - Tumour necrosis factor-alpha and macrophages in Plasmodium berghei-induced cerebral malaria. AB - The effect of tumour necrosis factor-alpha on malaria-infected mice was studied. C57Bl/6J mice infected with Plasmodium berghei K173 exhibited an increased sensitivity to exogenous TNF. Injection of 15 micrograms TNF was lethal to some of the animals when given 5-7 days after infection, while when given later on in the infection (i.e. days 8-10) amounts as low as 2.5 micrograms TNF appeared to be lethal in all mice. The pathology in infected mice treated with TNF resembled that found in the brains of infected mice dying with cerebral malaria. Infected mice treated with TNF, however, also developed severe pathological changes in other organs. On the contrary, treatment with sublethal amounts of TNF (1.0 micrograms or less) given on days 8 and 9 after infection, protected mice against the development of cerebral malaria. In addition, infected mice exhibited and enhanced sensitivity for treatment with lipopolysaccharide (LPS). Sublethal amounts of LPS, however, did not prevent mortality as in TNF-treated mice (LPS treated mice died at about the same time as infected mice that developed cerebral malaria), but no cerebral haemorrhages were found in the majority of LPS treated, infected animals. Treatment with dexamethasone during infection protected mice against the development of cerebral malaria, but did not suppress their increased sensitivity to exogenous TNF. Treatment of mice with liposome-encapsulated dichloromethylene diphosphonate (lip-Cl2MDP), used to eliminate macrophages (an important source of TNF), prevented the development of cerebral malaria, but only when given before day 5 of infection. Mice protected by treatment with lip Cl2MDP, however, remained sensitive for LPS on the eighth day of infection. PMID- 8414667 TI - Detection and characterization of DNA polymerase activity in Toxoplasma gondii. AB - A DNA polymerase activity has been detected and characterized in crude extracts from tachyzoites of Toxoplasma gondii. The enzyme has a sedimentation coefficient of 6.4 S, corresponding to an approximate molecular weight of 150,000 assuming a globular shape. Like mammalian DNA polymerase alpha, the DNA polymerase of T. gondii was sensitive to N-ethylmaleimide and inhibited by high ionic strength. However, the enzyme activity was not inhibited by aphidicolin which is an inhibitor of mammalian DNA polymerases alpha, delta and epsilon and also cytosine beta-D-arabinofuranoside-5'-triphosphate which is an inhibitor of alpha polymerase. The activity was inhibited by 2',3'-dideoxythymidine-5'-triphosphate which is an inhibitor of mammalian DNA polymerase beta and gamma. Magnesium ions (Mg2+) were absolutely required for activity and its optimal concentration was 6 mM. The optimum potassium (K+) concentration was 50 mM and a higher concentration of K+ markedly inhibited the activity. Activity was optimal at pH 8. Monoclonal antibodies against human DNA polymerase alpha did not bind to DNA polymerase of T. gondii. Thus the T. gondii enzyme differs from the human enzymes and may be a useful target for the design of toxoplasmacidal drugs. PMID- 8414668 TI - Rickettsia-like organisms and chitinase production in relation to transmission of trypanosomes by tsetse flies. AB - Rickettsia-like organisms (RLO) from testse midguts and mosquito cell cultures showed high levels of endochitinase activity. A line of Glossina morsitans morsitans highly susceptible to midgut trypanosome infection and with high incidence of RLO infection showed significantly greater chitinolytic activity than G. austeni which had low RLO incidence and were correspondingly refractory to midgut infection. Midgut infection rates of Trypanosoma brucei rhodesiense in G. m. morsitans showed a dose-related increase when flies were fed N-acetyl-D glucosamine (GlcNAc) in the infective meal and for 4 subsequent days. A model is proposed for susceptibility to trypanosome infection based on the generation of GlcNAc by RLO endochitinase activity in tsetse pupae inhibiting midgut lectin in teneral flies. PMID- 8414669 TI - Small subunit (18S) ribosomal RNA gene divergence in the genus Schistosoma. AB - An entire 18S rRNA gene sequence from Schistosoma spindale (1990 bases) and partial 18S rRNA gene sequences from S. haematobium (1950 bases) and S. japonicum (1648 bases) have been determined. Together with the previously published sequence of the S. mansoni 18S rRNA gene, these data encompass the 4 recognized Schistosoma species groups. Although Schistosoma 18S rRNA genes are highly conserved, the sequences permit a preliminary molecular phylogeny to be established for the genus. This identifies S. haematobium and S. spindale as sister taxa in a clade with S. mansoni. S. japonicum does not appear to be closely related to this clade. Much of the observed variation occurs within a 'hypervariable' stretch of the gene corresponding to part of the V4 region of 18S rRNA. Despite this variation, the 3 new sequences fit models of 18S rRNA secondary structure predicted from the S. mansoni sequence. PMID- 8414670 TI - Discrimination between six species of Theileria using oligonucleotide probes which detect small subunit ribosomal RNA sequences. AB - The complete small subunit ribosomal RNA (srRNA) gene of Theileria parva was cloned and sequenced. Two primers were designed which permitted the specific amplification of part of the Theileria srRNA gene from Theileria-infected cell line samples which were predominantly (> 95%) bovine DNA. The sequence of the central (variable) region of the srRNA genes of T. annulata, T. taurotragi, T. mutants and two unidentified parasites referred to as Theileria sp. (buffalo) and Theileria sp. (Marula) were obtained. An alignment of the sequences was generated from which 6 oligonucleotide probes, corresponding to species-specific regions, were designed. These probes were demonstrated to provide unequivocal identification of each of the 6 species either by direct detection of parasite srRNA or by hybridization to amplified parasite srRNA genes. The probes were not able to distinguish buffalo-derived T. parva, the causal agent of Corridor disease, from cattle-derived T. parva, the causal agent of East Coast fever. PMID- 8414671 TI - Epidemiology of Toxocara canis in red foxes (Vulpes vulpes) from urban areas of Bristol. AB - A descriptive epidemiological survey was undertaken of the ascarid nematode Toxocara canis in 521 red foxes (Vulpes vulpes) during the period January 1986 to July 1990. Age-prevalence and age-intensity profiles show that worm burdens are significantly higher in cubs than in subadult or adult foxes and higher in subadult than in adult foxes. Seasonal variations in worm burdens occur, with the highest prevalences and intensities being found during the spring, when cubs are born, and in the summer months. Prevalences and intensities then decrease during the autumn and winter months in both subadult and adult foxes, but, during this period, prevalences are significantly higher in male than in female foxes. Variations in worm burdens in the fox population are likely to be related to the reproductive cycle of the fox, with a high proportion of cubs becoming infected in utero. The role of the fox in the transmission of T. canis in the urban environment is discussed. PMID- 8414672 TI - Viability of Onchocerca volvulus in vitro. AB - The use of a selective schedule of tests to identify a viable population of isolated adult Onchocerca volvulus (Nematoda: Filarioidea) has been investigated in a large worm population. The study was initiated to develop methodology appropriate to test new candidate macrofilaricides for their in vitro activity against O. volvulus. After removal from the host the viability of isolated intact parasites was estimated by assessing the motility indices of male worms, and the colorimetric quantification of the reduction of the bioreducible tetrazolium reagent XTT and lactate output by female worms. Additionally the motility of whole females and the movement of inner organs of female worms were scored quantitatively. These response parameters were used to sort the adult worms into viability groups at the start of the in vitro culture. The adult worms were then observed for 6 days and viability was assessed regularly during the culture period. At the end of the culture period, the reduction of the water-insoluble tetrazolium reagent MTT was used to determine the formazan formed by the entire male and female worms. The response parameters used at the start of the culture proved to be highly predictive for detecting viable and non-viable adult worms. In the group of worms selected as 'viable' around 70% kept their motility and metabolic activity at a high level until the end of the culture compared to the initial level. In contrast, none of the female worms and only 13% of the male worms categorized as 'poorly viable' demonstrated a motility index or metabolic level at the end of the culture period that was comparable to that of the worms in the 'viable' groups. For female worms the lactate output correlated significantly with weight whereas no correlation was seen between MTT reduction and weight. PMID- 8414673 TI - Social behaviour, stress and susceptibility to infection in house mice (Mus musculus): effects of duration of grouping and aggressive behaviour prior to infection on susceptibility to Babesia microti. AB - Unrelated and initially unfamiliar male CFLP mice, maintained for different periods in groups of 6, differed in both their rate of clearance of Babesia microti and the time taken to reach peak parasitaemia in relation to their aggressive behaviour within groups prior to infection. Males maintained in groups for shorter periods and showing more aggression within their group were slower to clear infection and males showing more marked external evidence of aggressive interaction reached a peak of parasitaemia sooner. Serum IgG and corticosterone analyses were consistent with increased aggression causing stress-induced immunodepression but relationships with aggression and social status were not simple. Males showing more aggression tended to enter their groups with higher levels of corticosterone and, to a lesser extent, reduced levels of IgG compared with other mice. The results thus suggest that increased susceptibility to disease may be a cost to males aggressively maintaining high social status. PMID- 8414674 TI - Caecal threadworms Trichostrongylus tenuis in red grouse Lagopus lagopus scoticus: effects of weather and host density upon estimated worm burdens. AB - Trichostrongylus tenuis eggs were counted in faeces from individually marked wild red grouse for 8 years. Egg counts varied seasonally and annually. In some years, a sudden increase in mid-April was consistent with delayed maturation of larvae which had overwintered in the birds in a hypobiotic state. A more gradual increase in summer was probably due to uninterrupted maturation of larvae ingested then. Despite 30-fold year-to-year variation in mean egg counts, relative differences in egg counts among known individuals within years tended to persist across years. Rainfall in previous summers explained much of the year-to year variation in egg counts, probably because parasite recruitment was greatest during wet summers. Grouse density was only weakly related to worm egg counts. The data were not consistent with the hypothesis that the cyclic-type population fluctuation in red grouse numbers observed at the time of this study was caused by the parasites. PMID- 8414675 TI - Inhibition of T cell receptor dependent activation pathways by HIV-1. PMID- 8414676 TI - [Cytokines and HIV infection]. PMID- 8414677 TI - [In vivo regulation of c-myc proto-oncogene: contribution of transgenic mice]. PMID- 8414678 TI - [Brucellosis among occupationally exposed people: evaluation of immunological tests after vaccination]. AB - For the evaluation of immunological tests during an epidemiological survey and of vaccination with the PI brucellin vaccine, in an occupationally exposed environment, a sample group of 354 subjects was studied. The vaccinal strategy was based on the outcome of a skin test for hypersensitivity: the PS brucellin test. In this framework, the serological status and evolution of individuals with positive or negative reactions to this test were analysed. Sera were studied using the buffered antigen test, indirect fluorescence immunoassay and Wright's agglutination test, as well as by PACIA and ELISA techniques with assay of IgG, IgA and IgM antibodies. The PS test, which was pivotal in this study, was compared with the lymphoblastic transformation test. One prominent aspect of this evaluation was the establishment of conventional prognostic indices for the PS and serology. The PS is definitely shown to be a convenient, reliable tool for screening. Although it does not generate sensitivity it may modify the serological status of both positive and negative individuals. PMID- 8414679 TI - [Thyrotropin assay by chemiluminescence in the diagnosis of dysthyroidism with low thyrotropin and normal thyroid hormones levels]. AB - The performances of a new "3rd generation" chemoluminescence TSH assay (TSH ICMA) with a functional sensitivity of 0.005 mU/l were compared with those of an "ultrasensitive" TSH immunoradiometric assay (TSH IRMA) in a series of patients characterised by a TSH IRMA less than 0.20 mU/l and normal free thyroxin (T4 L) and triiodothyronine (T3 L) concentrations. The 95% cut-off value for hyperthyroidism was 0.03 for TSH ICMA and 0.05 for TSH IRMA. In a first group of 41 subjects undergoing Tc99m thyroid scan, images of multifocal increased uptake or toxic adenoma were associated with a lower TSH ICMA than in patients with a normal isotope scan. TSH ICMA was also lower than TSH IRMA (p < 0.01). At the cut off value of 0.03 mU/l, the specificity of TSH ICMA was higher than that of TSH IRMA, but the sensitivity were identical. In a second group of 36 patients with severe non-thyroid diseases, TSH ICMA was lower than the cut-off value for hyperthyroidism in 30% of cases, while TSH IRMA was lower than the cut-off value in 40% of cases. A satisfactory concordance was observed between the two methods. In conclusion, the two TSH assays, IRMA and ICMA, provide globally comparable information in subjects with a low TSH and normal T4 L and T3 L. However, the better specificity of TSH ICMA and a smaller overlap with the frank hyperthyroid zone in patients with non-thyroid disease argue in favour of the use of this new assay method. PMID- 8414680 TI - [In vitro pharmacological study of fluconazole. Evaluation of an antifungal assay]. AB - Fluconazole is a new antifungal drug which is now very often used for the treatment of yeasts infections: mucocutaneous candidosis, systemic candidosis and cryptococcosis. Some resistance seems to occur in clinical practice. It turns out now that it must be necessary to test the drug susceptibility in vitro of the yeast strains isolated from patients. Unfortunately, a current laboratory technique to test this agent doesn't exist. In this work a useful method for Minimal inhibitory concentration (MIC) evaluation in a commercial available broth medium, and a technique for testing fluconazole disks on an agar medium are studied. 150 strains of different yeasts species are tested. A regression curve is performed to correlate the two methods. PMID- 8414681 TI - [Changes in slow and fast electrical activity of the gastro-duodeno-jejunal junction after cholecystectomy in humans]. AB - The re-establishment of regular gastrointestinal electrical activity (Migrating Myoelectrical Complex-MMC- and slow wave) after cholecystectomy for gallstones was investigated in six female patients (50-77 years) by means of serosal electrodes implanted in the wall of the antrum, duodenum and jejunum during surgery. Five hour recordings were made on the first, third and fifth postoperative days, in patients who were on a liquid diet. The recordings were made after a twelve hour fast. On the fifth postoperative day, a test meal (250 g yoghurt) was given to the patients ant its effects on electrical activity were monitored for 2 hours. Even though MMC were present on the first and third postoperative days, a detailed study of their origin, the length of the Phase 3 and the speed of gastro-jejunal propagation showed that there existed an inhibition of gastric MMC until the fifth postoperative day and that intestinal MMC was slower than normal until the third postoperative day. On the fifth postoperative day, gastric inhibition disappeared since the length of the Phase 3 of the MMC of the stomach corresponds to those of the duodenum and jejunum, and the speed of propagation corresponded to that in normal subjects. Further, the slow wave frequency peculiar to each segment studied increased progressively from the first to the fifth postoperative day and the same was true of the slow waves with superimposed action potentials. In conclusion, notwithstanding the presence of MMC from the first postoperative day, it was only on the fifth day that normal coordination was restored and hence that patients were able to eat again. PMID- 8414682 TI - [Changes in plasma lipoproteins in adolescents with trisomy 21 in response to a physical endurance test]. AB - In a previous study, the authors recorded a free fatty acids (FFA) concentration decrease in 11 Down's Syndrome (DS) subjects after an incremental maximal exercise until exhaustion. The aim of this study was to determine if lipid metabolism parameters in a group of DS subjects could be changed after an endurance training period prior to a sustained physical exercise test. After an "adapted" exercise programme, 6 healthy DS subjects, 4 boys and 2 girls aged from 16 to 22 years, performed a sustained exercise lasting 40 minutes on an cycle ergometer. They also compared their values with those taken from the data of laboratory for similarly aged normal subjects. The results of this trial indicate: (a) at rest: (i) a lipid metabolism that included normal level of triglycerides (Trig), but low level of total cholesterol (TC), (ii) a pathological pattern of lipoproteins: low level in high density lipoprotein (HDL), and high level in pre beta very low density lipoprotein (VLDL); (b) after an "adapted" exercise programme period concluding with a final test: (i) an approximate adjustment to normal lipid profiles: rise in HDL and fall in VLDL (ii) a rise of free fatty acids (FFA) between the start and the end of the test. It is noted that physical activity of an endurance type appears to have a favourable long term effect on some manifestations of this genetic disease and that, consequently, such endurance training may be promoted. PMID- 8414683 TI - [Human cytomegalovirus infection: new methods for virological diagnosis]. AB - Recently developed methods have greatly increased the sensitivity and speed of virological diagnosis of cytomegalovirus infection. Virus can be detected in infected cell cultures within 24 or 48 hours of specimen inoculation by using monoclonal antibodies to immediate-early antigens in immunocytochemistry procedures or DNA sequences in hybridisation in situ assays. CMV antigens can also be detected directly in infected cells within clinical specimens. An early antigen can be visualized in nuclei of circulating leukocytes from viremic patients. DNA hybridization is used for CMV analysis in Dot-blot, Southern-blot and in situ hybridization assays. DNA amplification, by polymerase chain reaction (PCR), has proven to be a very sensitive method for diagnosis of CMV infection and should be useful for investigation of CMV pathogenesis and latency. Serologic assays such as ELISA and latex agglutination assays are accurate for screening donors and recipients of blood and organ or marrow graft. Studies of viral protein epitopes recognized by human sera are in progress. PMID- 8414684 TI - [Soluble antigens of intra-erythrocyte stages of Plasmodium falciparum: diagnostic and vaccinal value]. AB - A wide variety of soluble antigens also called exoantigens or excretory-secretory antigens are released by asexual blood stages of P. falciparum at the time of schizont burst and subsequent merozoite invasion. At present, most of these soluble malarial proteins have been purified and have had their genes cloned; the primary sequence displays considerable polymorphism upon tandem repeats. However, its function and the relevance of the polymorphism for the induction of host immune response, predominantly IgM and T-cell-independent type, has yet to be determined. The potential of P. falciparum exoantigens as immunodiagnostic tools has been the focus of numerous studies. Enzyme immunoassays or radioimmunoassays for the detection of malarial antigens in blood of suspected peoples have been reported. Their development requires increasing both specificity and sensitivity. Recently, some P. falciparum exoantigens have been proposed as candidates for inclusion in an anti-disease vaccine which induces a clinical but not parasiticidal immunity against malaria. There is much interest in this concept and their efficiency remains to be confirmed, as new strategies are needed in the prevention of malaria. PMID- 8414685 TI - [Infectious complications of venous catheters]. AB - Infection is the most frequent complication of venous catheters. The various types of infection are defined, followed by a review of the modalities of contamination of a catheter. The microorganism most frequently involved is Staphylococcus. The bacteriological diagnosis of a venous catheter infection is essentially based on semiquantitative culture according to Maki's technique or a quantitative method (Cleri or Brun-Buisson). Bacteriological diagnostic methods are also being developed to allow maintenance of the catheter. Treatment is primarily preventive, while, in addition to systemic antibiotics, curative treatment may also consist of attempts to maintain the infected catheter in place and the so-called "antibiotic lock" technique. PMID- 8414686 TI - A pediatrician's view. The eyes have it. PMID- 8414687 TI - Neonatal conjunctivitis. PMID- 8414688 TI - Acute conjunctivitis of childhood. PMID- 8414689 TI - Treatment of conjunctivitis in infants and children. PMID- 8414690 TI - Chronic conjunctivitis in infants and children. PMID- 8414691 TI - Serious eye infections in children. PMID- 8414692 TI - A pediatrician's view. Collaboration between pediatric subspecialists and generalists in the care of HIV-infected children. PMID- 8414693 TI - The epidemiology of pediatric HIV-1 infection. AB - While the incidence of HIV infection has leveled off in older homosexual men, infection in women, children, and adolescents is rising. Unless effective intervention measures are implemented, the number of pediatric patients with HIV and related illnesses will continue to increase. Interventions aimed at curtailing perinatal transmission potentially will have the greatest impact on decreasing the incidence of pediatric AIDS. Studies evaluating strategies aimed at this are currently underway. Educational initiatives targeting adolescents also may lower the number of pediatric patients with HIV infection. As treatments improve, the majority of HIV-infected children will survive for years and perhaps decades. The number of children with HIV infection in the upcoming decades will have a profound impact on our communities and our practice of pediatric medicine. PMID- 8414694 TI - HIV infection in pregnancy. PMID- 8414695 TI - Maternal-fetal transmission of HIV-1 infection. AB - Studies to examine the mechanisms responsible for perinatal HIV transmission and clinical trials to reduce this transmission are ongoing. A more complete understanding of the timing and modalities of vertical transmission will permit more precise targeting of future clinical trials. Perinatal transmission of HIV infection is most likely due to multiple factors. The degree of viral replication, neutralizing antibody and other maternal immune factors, genetic factors, and an intact placental barrier are most likely important and may be interrelated. Multiple different combination therapies may be necessary to interrupt maternal-fetal HIV transmission. Pregnancy is a well-defined and limited time period, and offers a model for HIV transmission where clinical, virological, and immunological factors can be evaluated in relation to the occurrence of HIV infection in the infant. The analysis of perinatal trials may provide insight into factors involved in protection and have implications. As HIV infection rates in the heterosexual populations of US inner cities and in impoverished areas of the third world continue to rise rapidly, strategies are needed urgently to curb the AIDS epidemic in children. Targeting the HIV-infected pregnant woman for therapies is one of the few hopes we have of preventing infection in children at risk of HIV infection. Without such therapeutic strategies, many of these at-risk infants are destined to acquire a chronic viral infection with significant morbidity and mortality. PMID- 8414696 TI - Clinical aspects of HIV infection in children. PMID- 8414697 TI - Current management of HIV infection in children. PMID- 8414698 TI - HIV and adolescents. AB - Human immunodeficiency virus has entered the adolescent population, and pediatricians will be caring for youngsters who are HIV positive or at risk of being infected. We should remember that it is risky sexual behavior and injection drug use that places a teen at risk for HIV infection, not their sexual orientation, ethnicity, or gender. Parents and their teenagers desire and expect their pediatricians to provide care and guidance for the diseases and problems that adolescents face. A screening psychosocial assessment of teenagers can identify those who need more in-depth counseling. Although the subjects of sex and drug use initially may be uncomfortable for a physician, practice using the HEADSS assessment will rapidly lead to comfort in discussing these important subjects. An understanding of HIV testing and pre- and post-test counseling will prepare the physician for the inevitable patient who wishes testing or who is HIV positive. Much of the treatment of HIV-positive adolescents involves patience and support while the adolescent grapples with the serious implications of being HIV positive. The initial history and physical exam establishes baselines regarding previous infections and illnesses that may bear on HIV infection, as well as determining which symptoms and signs of HIV infection are present. The initial laboratory tests further define the patient's current clinical state and will determine what therapies are immediately needed. Human immunodeficiency virus continues to evolve toward a manageable chronic illness that responds most favorably to early intervention. PMID- 8414699 TI - Cephalosporin properties and dosing. PMID- 8414700 TI - Evaluating the potential for serious bacterial infection in infants and young children. PMID- 8414701 TI - The prevalence of serious bacterial infections by age in febrile infants during the first 3 months of life. PMID- 8414702 TI - Defining fever and other aspects of body temperature in infants and children. PMID- 8414703 TI - Evaluation and management of the febrile infant 60 days of age or younger. AB - The old dogma of conservative management and hospitalization for all young febrile infants may no longer be necessary or considered the best option for many febrile infants encountered by practitioners today. All infants will continue to require individualized decisions regarding their care by astute clinicians well versed in the best options for the management of these children. The suggestions outlined in this article are meant to be a guide for the clinician to the available options for the evaluation and management of the febrile infant 60 days of age or younger. PMID- 8414704 TI - Occult bacteremia in the 3-month-old to 3-year-old age group. AB - Occult bacteremia precedes many serious infections in children. The vast majority of patients with occult bacteremia have an elevated temperature (> or = 39 degrees C), but fever is an extraordinarily common presenting complaint in the 3- to 36-month-old age group, which is at highest risk for S pneumoniae, H influenzae type b, and N meningitidis bacteremia. On examination, most patients with bacteremia will have no findings that distinguish them from nonbacteremic children. A white blood cell count of > 15,000/microL in a child with fever will identify about two thirds of children with occult bacteremia. Blood culture remains the most definitive test. It is important to establish a rational strategy to identify and treat these children before significant sequelae occur. PMID- 8414705 TI - Management of infants and children 3 to 36 months of age with fever without source. AB - The evidence and guidelines presented in this article are meant to assist clinicians who manage children with FWS. However, physicians may choose to individualize therapy based on unique clinical circumstances or to adopt a variation of these guidelines based on a different interpretation of the evidence concerning these issues. No guidelines can eliminate all risk nor confine antibiotic treatment only to children likely to have occult bacteremia. The optimal management strategy reduces risk to a minimum at a reasonable cost and can be used in most practice settings. PMID- 8414706 TI - Urinary tract infection in outpatient febrile infants and children younger than 5 years of age. PMID- 8414707 TI - Extracorporeal membrane oxygenation. Will this therapy continue to be as efficacious in the future? AB - The efficacy of ECMO has been discussed. If only a dichotomous live/die outcome is used as a measure of utility, ECMO is efficacious for infants with a greater than 20% mortality rate. Using a Bayesian approach and neonatal follow-up data, one concludes that ECMO, as used at present, is effective. Future measures of efficacy will vary depending on utility measures used, the populations studied, and the relative efficacy of alternative therapies. PMID- 8414708 TI - Neonatal and infant heart transplantation. AB - Since the advent of cyclosporine A, cardiac transplantation has been offered routinely for adolescents and children with cardiomyopathy refractory to medical management. Recently, cardiac transplantation has been offered to neonates with severe structural congenital heart disease. This article reviews the indications, pretransplant evaluation, and post-transplant management of neonates and infants undergoing cardiac transplantation. PMID- 8414709 TI - Necrotizing enterocolitis. New thoughts about pathogenesis and potential treatments. AB - Necrotizing enterocolitis (NEC) remains a major cause of morbidity and mortality in premature infants. An incomplete understanding of its pathogenesis has hampered efforts to devise an effective preventative strategy. New insights into the pathogenesis of NEC, particularly at the cellular and biochemical level, however, offer a rational basis for the development of new approaches to this disease. PMID- 8414710 TI - Hypoxic-ischemic brain damage in the term infant. Lessons from the laboratory. AB - Term infants who have sustained brain damage from intrauterine asphyxia can often be classified into one of two distinct clinical and radiologic syndromes. The first consists of damage to the cerebral hemispheres. The second consists of damage predominantly involving the basal ganglia and thalamus. Results of studies of asphyxia in experimental animals are presented. PMID- 8414711 TI - Screening for metabolic disorders. How are we doing? AB - Routine screening for phenylketonuria and congenital hypothyroidism has become an integral part of pediatric practice in the United States. Screening for several other metabolic disorders is now entering the second or third decade of use. New information is available for the pediatrician for both groups of disorders that will be of help in caring for children in the years to come. PMID- 8414712 TI - Tuberculosis in infancy in the 1990s. AB - In 1990, 25,701 cases of tuberculosis (TB) were reported in the United States, the largest annual increase since 1953. Children younger than 15 years of age accounted for 1596 new cases. The resurgence of TB can largely be contributed to the HIV epidemic. The clinical course, diagnosis, therapy, and prevention of TB in the perinatal period and in infancy are discussed in view of the epidemics of HIV and TB in the adult population. PMID- 8414713 TI - Advances in neonatology. View from a practicing pediatrician. AB - After reviewing the many problems that may be seen by the physician in follow-up care of the preterm infant and family, it is important to take a step back and evaluate the infant as a whole. In the vast majority of instances, the preterm infant will turn out to be normal. In an unfortunate minority, there may be difficult problems that the baby, parents, and caretakers must face. The pediatrician must remain diligent to attend to those problems that are correctable and to assist patients to their full potential. PMID- 8414714 TI - Apnea spells, sudden death, and the role of the apnea monitor. AB - SIDS is the most common cause of death between the ages of 1 week and 1 year. It affects 1 out of every 500 to 600 live births. The etiology of SIDS is unknown. There are no tests currently available to predict the infant who will die from SIDS prior to death. SIDS cannot be prevented. Many infants experience serious apneic spells, however, that require diagnostic evaluation and treatment. Even if the treatment of these infants does not have a large impact on the SIDS rate for the general population, thorough diagnostic evaluations and appropriate use of home apnea-bradycardia monitoring is indicated for this population and may reduce their risk of morbidity or mortality. PMID- 8414715 TI - Surfactant replacement therapy for pulmonary diseases. AB - Surfactant therapy has clearly been a meaningful addition to the therapeutic armamentarium in the management of premature infants with RDS. Pediatricians and others involved in the care of newborn infants should familiarize themselves with the various surfactant preparations, the indications for their use, the techniques of administration, and the possible side effects. All such care provides should also be skilled in endotracheal intubation and ventilation of neonates; recognition of the clinical and radiographic signs of RDS; and have the appropriate equipment to monitor cardiopulmonary status, oxygenation, and ventilation in these infants until transport to a tertiary care facility can be accomplished. In addition to the two current FDA-approved surfactants, several other surfactants are in various stages of evaluation. When administered to infants with established RDS, both natural and synthetic surfactants have clearly been shown to improve survival, decrease requirements for ventilatory support, and reduce the incidence of air leak complications. Although by no means conclusively demonstrated, certain infants, particularly those delivered at < 30 week gestation, may benefit from immediate treatment in the delivery room. It should be emphasized that, except under extenuating but controlled circumstances and except in the hands of an experienced physician, surfactant treatment should not be viewed as an integral part of neonatal resuscitation. Adequate treatment requires the administration of a minimum of two surfactant doses, although some infants may benefit from additional doses or treatment with an alternative preparation. Massive pulmonary hemorrhage, although rare, is observed with prophylactic and rescue treatment protocols and may result from hemorrhagic pulmonary edema due to a hemodynamically significant PDA. Currently there are no data to recommend the use of one surfactant preparation over another. The short- and long-term benefits may be similar with different products. Therefore, we must await results of trials with then necessary power (large number of subjects) and unbiased design to discern any clinically relevant differences. Results of studies directly comparing the relative efficacy of Survanta and Exosurf, conducted under the auspices of the National Institutes of Health, are expected in 1993. Multicenter trials comparing prophylactic and rescue administration of Exosurf versus CLSE and Survanta versus CLSE are currently underway. It is encouraging to note that follow-up studies up to 2 years of age do not reveal an increase in physical or neurodevelopmental handicaps, BPD, or other problems in preterm infants who received surfactant preparations either for prophylaxis or rescue therapy. Results of long-term follow-up studies, however, are not yet available.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8414716 TI - Developmental outcome in extremely premature infants. Impact of surfactant. AB - Advances in neonatology, particularly surfactant, have enabled us to significantly improve mortality in the extremely low-birth-weight prematurely born infant. The impact on morbidity remains less clear but decidedly optimistic as the preponderance of the data currently available suggests that although we have not improved outcome in this high-risk group of infants, neither have we increased the percentage of infants with significant impairments. But overall morbidity remains high, especially for the smallest and earliest survivors. There is a growing body of research that supports the idea that with modification of the stressful impact of the NICU environment on the physiologically unstable and vulnerable premature infant, we may be able to improve developmental outcome. We are already seeing even very small infants extubated at a younger age and ready for discharge home well before their originally anticipated "due date." Within this setting we have unequalled opportunity to positively impact development by caring for the infants in a supportive environment that integrates parents as primary caretakers who are comfortable and competent at the time of the infant's discharge home. Further research endeavors should be enlightening in this regard. PMID- 8414717 TI - Meconium staining and the meconium aspiration syndrome. Unresolved issues. AB - Meconium staining of the amniotic fluid and the meconium aspiration syndrome will likely remain common occurrences faced by health care providers. Unfortunately, our understanding of these entities is incomplete. There are a number of issues which need to be adequately evaluated regarding the pathophysiology of MAS, the delivery room management of the meconium-stained infant, and the neonatal intensive care unit management of MAS. Currently, there is much dogma in various hospitals throughout this country regarding these issues, dogma which has yet to be substantiated by quality scientific investigations. PMID- 8414718 TI - Advances in the treatment of persistent pulmonary hypertension of the newborn. AB - Despite an association with meconium and blood aspiration, pneumonia, sepsis, pneumothorax, prematurity, and congenital diaphragmatic hernia, no cause for persistent pulmonary hypertension of the newborn can be found in many cases. This article discusses the advances in current therapies including the promising new therapy of inhaled nitric oxide. PMID- 8414719 TI - Strength training and the immature athlete: an overview. AB - The developing musculoskeletal structures of the immature athlete are uniquely susceptible to injury, particularly at the physes. These growth plates are present in arm and leg bones, and some may not close until the late teen years. Early literature suggested that weight training might be inappropriate for these athletes. However, recent evidence suggests that, properly done, strength/resistance training may not only be safe, it may also help reduce the risk of injury for the young athletes. Nurses are often called upon to advise coaches of formal and community athletic programs, and need to know the underlying physiology of developing bone and muscle as well as the current recommendations related to training. PMID- 8414720 TI - Sports-related injuries in youths: prevention is the key--and nurses can help!. Interview by Gus A. Ostrum. AB - The number of American children participating in youth sports within and outside of school settings has increased dramatically in recent years, leading to an increased number of sports-related injuries that include both physical and psychological problems. In this interview, Temple University Sports Medicine Centers' Ted Quedenfeld emphasizes that prevention is of paramount concern among those involved in youth athletic participation, and that nurses need to play a key role in prevention. PMID- 8414721 TI - Bicycle safety programs: targeting injury prevention through education. AB - Children especially are prone to bicycle injuries, many of which are preventable. Data from an assessment of pediatric bicycle-related injuries at a major Southeastern Tennessee trauma center were used to support the need for a public awareness program about children's bicycle safety. Results of the assessment are reviewed and program planning, implementation, and evaluation are described. PMID- 8414722 TI - Comparison of two skin-level gastrostomy feeding tubes for infants and children. AB - Gastrostomy tubes have improved technically over the last 10 years. New to the market are skin-level devices, which are low-profile in design and avoid many problems of standard gastrostomy tubes. Two skin-level devices, the Button and the MIC-KEY, are appropriately designed for infants and children and are compared. PMID- 8414723 TI - A pediatric protocol for management of extravasation injuries. AB - The administration of bioactive agents through IV can lead to serious complications, including partial-or full-thickness skin loss, infection, nerve and tendon damage, or loss of limb. Children's Hospital of The King's Daughters (CHKD) implemented a protocol to uniformity identify and treat infiltrates in order to avoid these serious potential complications. PMID- 8414724 TI - School-based clinics and the pediatric nurse: an interview with Surgeon General designee M. Joycelyn Elders. Interview by Judith E. Vessey and Margo N. Swanson. AB - Among Dr. Joycelyn Elders' goals as Surgeon General is to work toward a national school-based clinic program. In this interview, she speaks with Pediatric Nursing regarding the implementation of school-based clinics in Arkansas, arguments for and against school-based clinics, the financial responsibility for these clinics, and the role of pediatric nurses in this segment of health care reform. PMID- 8414725 TI - Managing your risk of legal liability. AB - Pediatric practitioners face significant risk of liability and are becoming increasingly conscious of the need to manage their risks of legal liability. Providers must understand what constitutes negligence or malpractice and steps they can take to reduce their risks of legal liability. PMID- 8414726 TI - Technical update brief: the child with a chest tube. AB - Caring for a child with a chest tube is a nursing challenge. By following a logical system of assessment, the critical care nurse will be able to master the art of chest drainage with little difficulty. This update provides the quick assessment procedure to assure proper chest drainage. PMID- 8414727 TI - Clarifying roles and expectations in home care. AB - Home care nurses work in an entirely different environment from where they were trained. This unique situation requires understanding the difference in the nurse's role in the home, and establishing rules that promote a therapeutic environment. PMID- 8414728 TI - Pediatric management problems. Iron-deficiency anemia. PMID- 8414729 TI - Health behaviors of adolescent male football athletes. AB - Heart disease is the number one cause of death in the United States. Primary prevention efforts have focused on adult populations even though there is evidence that heart disease risk factors are sometimes present in early childhood. Due to the nature of the game, adolescent male football athletes may participate in behaviors that increase heart disease risk such as power weight training and no aerobic exercise, increased calorie and fat intake, and increased substance use. PMID- 8414730 TI - Heparin vs. saline for peripheral i.v. locks in children. AB - PURPOSE: To determine the efficacy of saline versus heparin flush solution to maintain peripheral i.v. locks in a pediatric population. METHOD: A prospective, randomized, double-blind design was used. A sample of 124 peripheral i.vs. were flushed with either saline or heparin in saline. Subjects were infants over 28 days of age and children. FINDINGS: The heparin and saline groups were comparable for total hours duration of the i.v. and for incidence of complications. CONCLUSIONS: Saline is efficacious in maintaining patency of peripheral i.v. locks in children over 28 days of age. PMID- 8414731 TI - Update on common allergic diseases. PMID- 8414732 TI - Balancing personal risk with professional responsibility: the case of AIDS. AB - Generally, nurses have an ethical and legal obligation to provide treatment to persons with AIDS. When it is unclear whether nurses are morally required to provide treatment, they should engage in moral analysis, discuss their concerns with their colleagues and administrators, work within their institution's framework to resolve their concerns, seek legal counsel, or as a last resort consider working in another practice setting. PMID- 8414733 TI - Several resources are available regarding the death experience. PMID- 8414734 TI - Acute lower respiratory infection in Argentinian children: a 40 month clinical and epidemiological study. AB - In a total of 1,003 children (805 inpatients and 198 outpatients) with acute lower respiratory infections (ALRI), clinical, social, and environmental data were analyzed. The major clinical entities were bronchiolitis, pneumonia, bronchitis, and laryngitis. The first two of these predominated in inpatients; pneumonia and bronchitis were more common in older children, while bronchiolitis was observed in infants. Respiratory rates of > 50/min. were more common in younger children and in cases with bronchiolitis and bronchitis. Retractions showed markedly less age-dependent variations and were present in all severe cases with different clinical diagnoses. Retractions alone or associated with cyanosis were the best indicators for severity of ALRI. Among outpatients, fever and wheezing were more common; inpatients were younger, more frequently malnourished, and from a lower socioeconomic level; family history of chronic bronchitis, crowding, and parental smoking also prevailed in this group. Family asthma and exposure to domestic aerosols was more common among outpatients. Prematurity rate (17 and 15%) of all ALRI cases was twice that of the general pediatric population and a significant difference existed between in- and outpatients under 6 months of age when perinatal respiratory pathologies predominated among inpatients. It is suggested to consider the need for assessing personal, family, and environmental risk factors in addition to clinical signs and symptoms when severe cases of ALRI are evaluated. PMID- 8414735 TI - Relationship between exposure to dust mite allergen and bronchial response to exercise in schoolchildren sensitized to dust mites. AB - We assessed the relationship between the exposure to dust mite allergens and a bronchial response to exercise in 8-year-old schoolchildren. Dust was collected from the mattresses of 1,291 children and the concentration of mite allergens was estimated by a commercially available ELISA test using monoclonal antibodies (ALK, Copenhagen) against the major allergens of Dermatophagoides pteronyssinus (Der pt) and Dermatophagoides farinae (Der f). A positive bronchial response to exercise (decrease of peak expiratory flow > or = 15% after exercise) occurred in 21 (22.6%) of 101 children sensitized to mite allergens (wheal size > or = 4 mm) and in 51 (4.8%) of 1,070 nonsensitized children. In the highest exposure groups (> 10 micrograms allergen/g dust), 15% of children sensitized to Der f and 20% of children sensitized to Der pt were responsive to exercise. Corresponding figures for the lowest exposure groups (< 0.4 micrograms allergen/g dust) were 11 and 28%, respectively. This negative finding may indicate that measurement of allergen concentration in mattresses does not reflect true exposure or alternatively that at the age of 8 years high exposure to dust mite allergens does not affect bronchial response to exercise in sensitized children. PMID- 8414736 TI - Spirometry in the asthmatic child: is FEF25-75 a more sensitive test than FEV1/FVC? AB - The use of spirometry in the assessment of children with asthma is taking on new importance with the realization that considerable airway obstruction may exist in the absence of clinically detectable abnormalities. There has been controversy over the value and relative sensitivity of various spirometry indices. This study evaluated the forced expiratory flow between 25% and 75% of vital capacity (FEF25 75), forced expired volume in 1 second (FEV1), and the ratio between the FEV1 and the forced vital capacity (FVC) in 100 asthmatic children aged 6-17 years, 29 of whom were wheezing at the time of the evaluation. All children with clinical wheezing had a FEV25-75 < 2 standard deviations (SD) below the mean (-2 SD), whereas 8 had a normal FEV1. The majority of the wheezing children had abnormalities of all 3 indices, whether expressed as < or = -2 SD or, in the case of the FEV1/FVC, arbitrarily taken as < 80%. Sixty-seven children of the entire study group had at least 1 abnormal spirometric index, but 38 of these had no clinical abnormalities. Twelve children had a reduced FEF25-75 as the only abnormality. These results suggest that FEF25-75 is a sensitive index of airway obstruction. PMID- 8414737 TI - Lung function in children and adolescents with tetralogy of Fallot after intracardiac repair. AB - We studied lung function in 41 patients, aged 6-27 years, 1-5 years after intracardiac surgical repair (ICR) of tetralogy of Fallot (TOF) and about 5 years after the establishment of the Pediatric Cardiac Center in Prague. The measurements included vital capacity (VC), total lung capacity (TLC), functional residual capacity (FRC), residual volume (RV), forced expiratory flows (FEF), specific airway conductance (SGaw), lung recoil pressure (Pst), and specific static lung compliance (SC1st). Single or multiple abnormal lung function parameters were found in 83% of patients. Lung function was not related to shunting operations prior to ICR, hemoglobin concentration, and hematocrit, and had no specific pattern. Pst at 100% TLC and 90% TLC declined with increasing age at ICR and at lung function testing, while SC1st rose, as did the ratio FRC/TLC. Fifteen patients were studied before and after surgery. Single or multiple lung function tests were abnormal in 93% before and in 84% after ICR. After ICR the ratios FRC/TLC and also RV/TLC, FEF at 25% VC, and FEF at 60% TLC were less frequently abnormal, while Pst at 100% TLC and at 90% TLC, as well as SGaw and TLC, were more frequently abnormal after ICR. The results indicated a regression of smaller airway obstruction and lung hyperinflation after ICR. The evolution of abnormally compliant (emphysematous) lungs with growth of the patients might be a sign of permanent sequelae of early lung damage from abnormal pulmonary hemodynamics. PMID- 8414738 TI - Gender related differences in airway tone in children. AB - The effects of gender, volume history, and inhaled atropine and isoproterenol on lung mechanics were assessed in 16 normal boys and 14 normal girls using lung volumes, flow-volume curves, and oscillatory resistances. Flows were measured from full and partial forced expiratory flow-volume curves. Six girls and 6 boys were studied before and after inhaled atropine, and 10 boys and 8 girls before and after inhaled isoproterenol. Girls demonstrated a significant increase in flows on full and partial curves with a deep inspiration [Vmax-partial 0.73 +/- 0.34 (SD) to Vmax-full 0.80 +/- 0.37 and 0.83 +/- 0.20 to 1.06 +/- 0.29 TLC/s in each group] and following inhalation of isoproterenol on the partial curves only (0.73 +/- 0.34 to 0.93 +/- 0.40 TLC/s). Boys showed a small but significant increase in Vmax with isoproterenol on full curves but not on partial curves. Following atropine, boys demonstrated a significant increase in Vmax on partial flow-volume curves (0.78 +/- 0.28 to 1.00 +/- 0.35 TLC/s) and a significant decrease in specific respiratory resistance (7.6 +/- 2.7 to 5.1 +/- 0.9 cmH2O/s), whereas girls had no such changes. These data suggest that boys have greater resting airway tone than girls and that this tone is less responsive to deep inspiration and isoproterenol independently, although a combination of isoproterenol and a deep inspiration will produce increased flows in boys. Atropine reduces airway tone predominantly in boys, suggesting that the increased resting airway tone in boys is partially mediated via the vagus nerve. PMID- 8414739 TI - Dexamethasone treatment suppresses collagen synthesis in infants with bronchopulmonary dysplasia. AB - Collagen is an essential component of connective tissue and is present in the pulmonary interstitium. Collagen deposition is known to increase in many acquired chronic diseases, including bronchopulmonary dysplasia (BPD). Urinary excretion of hydroxyproline has been used as a specific index of collagen synthesis. Many studies have demonstrated that dexamethasone therapy is associated with respiratory improvement in infants with BDP but the mechanism of this effect is not well understood. We postulated that in infants with BDP who receive dexamethasone, suppression of collagen synthesis may cause respiratory improvement. Therefore, we studied the effect of dexamethasone on respiratory status and urinary excretion of hydroxyproline in 14 ventilator-dependent infants with BDP. Infants received 0.5 mg/kg/day dexamethasone, tapered by half every 3 days to complete a 12 day course. Eleven of the 14 infants were extubated at a mean +/- SD of 8.7 +/- 4.9 days after starting dexamethasone. Mean urinary hydroxyproline/creatinine ratios at 3, 6, 9, and 12 days of dexamethasone therapy were significantly lower than the mean pretreatment value, but after discontinuation rapidly rose toward baseline values. Decreased urinary excretion of hydroxyproline indicates that dexamethasone suppressed collagen synthesis in these infants. We speculate that suppression of collagen synthesis reduced pulmonary inflammation and fibrosis, resulting in respiratory improvement. PMID- 8414740 TI - Pulmonary sarcoidosis in children: serial evaluation of bronchoalveolar lavage cells during corticosteroid treatment. AB - The clinical course of sarcoidosis in children has not been well defined. Eight children with symptomatic sarcoidosis included in this study underwent pulmonary function tests and bronchoscopy with bronchoalveolar lavage (BAL) before treatment and during steroid therapy. At the start of therapy, functional parameters, mostly dynamic lung compliance and lung transfer factor for CO, were impaired. This was associated with abnormalities of BAL cell populations: increased total cell number with a high proportion of lymphocytes, modifications of lymphocyte subpopulation with an elevated CD4+/CD8+ ratio, and enhanced ability of alveolar macrophages to release hydrogen peroxide. Although respiratory abnormalities seemed to be similar at the initial stage of sarcoidosis in children and adults, the course of the disease appeared to be different. Despite the absence of respiratory symptoms and disappearance of chest radiographic abnormalities on prolonged steroid treatment, we found slow improvement of pulmonary functions associated with persistence of BAL lymphocytosis and elevated CD4+/CD8+ ratios. However, the ability of alveolar macrophages to release hydrogen peroxide was significantly reduced after 6 months of steroid treatment, and it remained identical to the control group. Therefore, the evaluation of disease activity appears to be critical for therapy in pediatrics, and for this purpose studies of alveolar macrophage activation may be of particular interest. PMID- 8414741 TI - Maturation affects the maximal pulmonary response to methacholine in rabbits. AB - Maximal bronchoconstriction in normal adults produces only small reductions in pulmonary function; however, in normal infants severe airway obstruction limits testing to low agonist concentrations. In this study, maximal pulmonary response to methacholine was evaluated in 5 immature (1 month old) and 5 mature (6 months old) rabbits. Animals were anesthetized, paralyzed, and mechanically ventilated via a tracheostomy tube. Changes in pulmonary function were assessed from maximal deflation flow volume curves following inhalation of doubling concentrations of methacholine between 0.6 and 320 mg/mL. Following 320 mg/mL methacholine, the immature rabbits had a greater percent decline in forced vital capacity (FVC) than the mature animals (55 +/- 15% vs. 36 +/- 10%; P < 0.05). At 50% FVC, isovolume flows were measurable in the 5 mature rabbits, and 4 of 5 had plateaus in their dose-response curves. At the higher methacholine doses, only 1 of 5 immature animals had measurable isovolume flows because of the decrease in FVC. There was no significant difference between immature and mature animals in the methacholine dose required to decrease baseline flows by 50%. We conclude that in rabbits maturation affects maximal pulmonary response but not the sensitivity to methacholine. PMID- 8414742 TI - Airway responsiveness in infants: comparison of inhaled and nasally instilled methacholine. AB - Airway responsiveness of infants is evaluated during sleep and the infants inhale the bronchial challenge agent via the nasal airway. Since stimulation of the nasal airway may produce bronchoconstriction, it is unclear whether the observed response in the infants results from deposition of the aerosol in the lower airways or from stimulation of nasal receptors. Therefore, in 6 healthy infants we compared the changes in partial expiratory flow-volume (PEFV) curves produced by aerosol inhalation of methacholine and the changes produced by instillation of equivalent doses of methacholine liquid into the nares. Following aerosol, the peak expiratory flow and the flow at functional residual capacity decreased, PEFV curves became concave in shape, and the oxygen saturation (SaO2) decreased. The highest methacholine concentration inhaled by any infant was 1.25 mg/mL. In contrast to aerosol delivery, a maximal methacholine concentration of 10.0 mg/mL was instilled into the nares of all 6 infants without any change in maximal flow at functional respiratory capacity (VmaxFRC) or SaO2. There was a significant decrease in peak flow and flattening of the PEFV curves at higher lung volumes; however, the PEFV curve remained convex in shape at the lower lung volumes. The changes in the PEFV curve following nasal instillation of methacholine are consistent with an increase in nasal resistance and no change in the lower airways. We conclude that the bronchoconstriction observed following inhaled methacholine does not result from stimulation of nasal receptors. PMID- 8414743 TI - Association between pulmonary and gastric inflammatory cells on the first day of life in preterm infants. AB - It has been shown that inflammatory cells in the newborn lung are fetal in origin, whereas those in the amniotic fluid are maternal. In order to explore the relationship between fetal amnionitis and neonatal pneumonitis, we collected paired samples of gastric aspirate within 2 hours of birth, and bronchoalveolar lavage fluid within 24 hours of birth from intubated preterm infants. Leukocyte counts in bronchoalveolar lavage fluid correlated with the duration of membrane rupture (r = 0.68, P = 0.0001). There was a high degree of correlation between leukocyte counts in the two fluids (r = 0.86, P = 0.0001). The factors responsible for this association are unknown. PMID- 8414744 TI - Fractal and morphometric analysis of lung structures after canine adenovirus induced bronchiolitis in beagle puppies. AB - Acute viral respiratory infections are commonly associated with alteration in lung growth and with chronic obstructive disease. However, it is difficult to quantify these changes in lung function. We determined that the recently described techniques of fractal analysis gave additional information about the changes in lung function after viral illness compared to standard morphometric techniques. Fractal and morphometric parameters change with lung growth after acute infection with canine adenovirus type 2 (CAV2, n = 5) or no infection (controls, n = 6) in beagle puppies. Lung pathological studies showed areas of obliterative bronchiolitis and chronic small airways inflammation but no emphysema in the CAV2-infected puppies. Morphometric studies at approximately 236 days of age demonstrated accelerated lung growth in the CAV2-infected dogs as evidenced by significant increases in lung volume (VL) and internal surface area (ISA). Fractal analysis showed an increased fractal dimension (Df) of the alveolar perimeter length in the CAV2 group associated with increased growth that was similar to the percentage change in VL and ISA. These data suggest that a single infection with CAV2 in beagle puppies accelerates lung growth and increases the complexity (Df) of the alveolar structure. PMID- 8414745 TI - Unusual form of endobronchial Aspergillosis in a patient with cystic fibrosis. AB - The isolation of Aspergillus fumigatus from airway secretions from patients with cystic fibrosis (CF) is common and usually denotes asymptomatic colonization or allergic broncho-pulmonary aspergillosis (ABPA). A 12-year-old boy with CF acutely developed moderately severe symptoms of unremitting cough, fever, dyspnea, weight loss, and cyanosis. Chest radiographs demonstrated widespread unilateral infiltrates and volume loss. By bronchoscopy tenacious mucous plugs were seen occluding the left lower lobe bronchus. Cultures from sputum and sequential bronchoalveolar lavage grew Aspergillus fumigatus, but other significant criteria for diagnosing ABPA were lacking. No improvement was seen with a 3 week course of systemic corticosteroid and antibiotic therapy. Treatment with amphotericin B and short-term mechanical ventilation resulted in rapid resolution of all symptoms. This form of endobronchial aspergillosis has not been described previously. PMID- 8414746 TI - New associations of primary ciliary dyskinesia syndrome. AB - Ten of 32 patients with primary ciliary dyskinesia syndrome (PCDS) also had other conditions. Five esophageal problems, 4 congenital heart disease, 2 scoliosis, and 4 miscellaneous and probably coincidental conditions were discovered. Additionally, a patient in this series, who has a normal heart, had a brother who died after surgery for complex congenital heart disease. In retrospect, he too probably had PCDS. The association of severe esophageal and cardiac disease with primary ciliary dyskinesia has not been described before. The diagnosis of PCDS may carry more implications than previously recognized. PMID- 8414747 TI - John A. Kirkpatrick, Jr.: a biography. PMID- 8414748 TI - Cervical neuroblastoma in eleven infants--a tumor with favorable prognosis. Clinical and radiologic (US, CT, MRI) findings. AB - Cervical neuroblastoma, a disease primarily of infants, has a favorable prognosis. Eleven patients are reported. Clinical presentations (other than mass) included stridor and swallowing problems. Masses when felt were commonly mistaken for infectious adenitis. Imaging studies (US, CT, MRI) showed solid masses with vascular displacement and narrowing; intraspinal extension was absent though extension into the adjacent sites of mediastinum and skull occurred. Horner syndrome was seen in five patients with accompanying heterochromia iridis in one. Five tumors had calcification. A high index of suspicion will lead to biopsy and less delay in diagnosis once a mass is felt or imaged. PMID- 8414749 TI - Cystic retrocerebellar malformations: unification of the Dandy-Walker complex and the Blake's pouch cyst. AB - Twenty-six cases of developmental retrocerebellar cyst (RCC) formation are studied with respect to determining the usefulness and anatomic relevance of separate terms currently in use, including Dandy-Walker complex, Dandy-Walker malformation, Dandy-Walker variant, mega-cisterna magna (MCM), and Blake's pouch cyst. An anatomic and embryological continuum between Dandy-Walker complex and Blake's pouch cyst is proposed. A method for the useful assessment of RCC is outlined. The patency or closure of the aqueduct is crucial to the evaluation and management of hydrocephalus associated with RCC formation. PMID- 8414750 TI - Assessment of pediatric near-drowning victims: is there a role for cranial CT? AB - Previous studies have suggested that CT examinations of the brain in children soon after near-drowning incidents are not helpful in predicting clinical outcome and are not necessary. The clinical and CT findings of 19 pediatric near-drowning victims were reviewed for correlation with clinical and neurologic outcome. As expected, a normal initial CT scan was poorly predictive of outcome, whereas an abnormal CT scan in the initial 36 h following an immersion incident was associated with a dismal prognosis. Three children with abnormal initial CT examinations were identified and all died within 3 days of admission. A CT scan performed in the immediate near-drowning period, therefore, may be helpful in identifying some patients who have sustained severe neurologic injury. PMID- 8414751 TI - Sonography of the knee in normal and abused infants. AB - Sonography of an infant's knee is relatively easy and can show the non-ossified cartilage and adjacent soft tissues with great clarity. The normal anatomic features are characteristic and easy to learn and recognize. In abused infants, sonograms can confirm subtle or questionable radiographic abnormalities. In infants with metabolic bone disease, detection of occult fractures of the epiphyses is clinically important. Metaphyseal abnormalities due to neonatal ricktes should not be mistaken as signs of child abuse. PMID- 8414752 TI - Acute experimental neuronal injury in the newborn lamb: US characterization and demonstration of hemodynamic effects. AB - BACKGROUND AND PURPOSE: Microinjection into the brain with N-methyl-D-aspartate (NMDA), a synthetic analogue of glutamate, has been used as a chemical model of perinatal hypoxic-ischemic injury. Little is known about the sonographic characteristics and hemodynamic consequences of these cytotoxic lesions. An understanding of these features may be useful in the early sonographic identification of stroke in newborns. METHODS: Twenty newborn lambs were anesthetized, paralyzed, and mechanically ventilated. Between 0.5 and 5 mu mole NMDA in 0.2 ml phosphate buffered saline, n = 18), or buffered saline only (n = 2) was injected into the right putamen under sonographic guidance. Serial grey scale and color Doppler images of the brain, Doppler spectra of the middle cerebral and thalamostriate arteries, cerebral blood flow (CBF) determinations using radiolabeled microspheres (n = 9), and cerebral oxygen extraction (n = 4) were obtained before, and at 15.60, and 120 min after NMDA injection. Pathologic examination was obtained in 11 animals. RESULTS: Homogeneous, well defined, moderately echogenic lesions surrounded by marked focal hyperemia on color Doppler were identified in every animal injected with 5 mu mole NMDA within minutes of injection. Lesions were characterized by focal areas of chromatolysis and cytoplasmic shrinkage, with scattered petechial hemorrhage. No lesions or hyperemia were observed in the animals injected with normal saline. Mean supratentorial CBF increased from 64 +/- 9 ml/min/100 g (control) to 152 +/- 30, 115 +/- 19, and 102 +/- 8 ml/min/100 g at 15, 60, and 120 min after injection respectively. The most marked increases occurred in right midbrain (467% of control), diencephalon (388%), and temporal lobe (282%), but were also observed in homotopic regions of the left hemisphere, and in pons, medulla, and cerebellum. Mean blood flow velocity in the middle cerebral artery, and thalamoperforator artery correlated well with changes in hemispheric and midbrain. CBF respectively. (r = 0.57-0.74, p = 0.0001, and r = 0.65-067, p = 0.0001 respectively). CONCLUSIONS: Focal brain lesions may by identified by sonography within minutes after experimentally induced neuronal injury. Alterations in echotexture are primarily due to intracellular cytoplasmic changes and microscopic hemorrhage. Local intracerebral injection of NMDA in newborn lambs increases both local and global CBF. PMID- 8414753 TI - MIBG detection of hepatic neuroblastoma: correlation with CT, US and surgical findings. AB - Metaiodobenzylguanidine (MIBG) imaging is used in the diagnosis, staging and follow-up of virtually every case of neuroblastoma seen at our institution. Normal sites of MIBG uptake include the liver and therefore difficulties have been predicted and encountered in the diagnosis of hepatic neuroblastoma due to inability to separate abnormally increased tracer deposition from normal hepatic activity. We reviewed every MIBG (I123 and I131) study performed at our pediatric hospital over a 4 year period encompassing 88 patients, 67 of whom had biopsy proven neuroblastoma. Hepatic findings onMIBG studies were compared with concurrent abdominal CT and US studies in all 67 patients. The clinical records of all patients with abnormal MIBG scans or abnormal CT or US studies of the liver were also reviewed. Eight patients were found to have abnormal liver findings on one or more imaging studies (MIBG, CT, or US). There were 3 true positive MIBG studies, one of which was an early study in a patient who later went on to have one of the false positive studies. Two patients had false positive MIBG scans for liver neuroblastoma. MIBG failed to detect liver involvement in 4 patients. PMID- 8414754 TI - Rational use of CT in acute pyelonephritis: findings and relationships with reflux. AB - Enhanced renal CT scanners were performed in 38 children (82% girls) to rule out acute pyelonephritis. Patients were divided in 2 groups on the basis of clinical presentation and bacteriology data. In patients of group A (n = 16, preliminary study), upper urinary tract infection (UTI) was certain. CT confirmed the diagnosis in all but 3 patients (a 2-year-old child and 2 patients with UTI developed on prior obstruction). In subsequently studied patients of group B (n = 22), clinical findings or bacteriology data were negative or questionable. CT made the diagnosis of acute pyelonephritis in 11 patients. As well as DMSA scintigraphy, CT scanner can help to diagnose or to rule out upper UTIs in difficult cases. In all boys of both groups, ipsilateral vesico-ureteric reflux (VUR) was found by subsequent voiding cystourethrography (VCUG) on the side of pyelonephritis. In girls, this correlation was shown in only 7 of the 25 kidneys with pyelonephritis. This result supports the hypothesis of a gender-dependent contamination. We believe that absence of radiologic reflux cannot exclude the possibility of bacterial crossings of ureteric meatus capable to lead to genuine upper UTIs. PMID- 8414755 TI - Renal abnormalities in children with hypertrophic pyloric stenosis--fact or fallacy? AB - Retrospective review of the abdominal ultrasound (US) examination of 274 children studied for hypertrophic pyloric stenosis (HPS) was undertaken to determine if there is an increased incidence of renal disease as previously reported. Five major abnormalities were detected in the 126 children with HPS. Three lesions were newly diagnosed and two had been diagnosed previously. Five children had abnormalities classified as minor or normal variants. Renal abnormalities were found in six of the 148 children who did not HPS. Only three of these were newly diagnosed and medically important. Eight children without HPS had minor abnormalities or normal variants of the kidneys. Newly diagnosed medically important renal lesions were present in 2.4% of children screened for HPS. The incidence of the finding was the similar in children with and without HPS. PMID- 8414756 TI - Perforation associated with barium enema in acute appendicitis. AB - A child with the suspected diagnosis of atypical acute appendicitis underwent a diagnostic barium enema. The study was complicated by perforation with leakage of a large amount of barium into the peritoneal cavity. The complex hospital course that resulted has prompted us to reevaluate the barium enema in the diagnosis of appendicitis and review the literature for contraindications. We conclude that this particular complication is extremely rare and that barium enema still has a part to play in some patients where the clinical diagnosis is uncertain. PMID- 8414757 TI - Retained fetal lung liquid in congenital lobar emphysema: a possible predictor of polyalveolar lobe. AB - The purpose of this study was to determine whether retention of fetal lung liquid is more prevalent in polyalveolar congenital lobar emphysema than in conventional congenital lobar emphysema. Two patients with congenital lobar emphysema were prospectively identified in a 3-year period. Twenty-five such patients were identified in a retrospective study covering 39 years. Medical records were available for 22 patients who had 23 emphysematous lobes. Both babies from the prospective study and six subjects from the retrospective group had respiratory symptoms and underwent chest X-ray in the first day of life. Six of the eight babies with respiratory symptoms and chest imaging in the first day of life had retention of fetal lung liquid in an emphysematous lobe. All six of these lobes were polyalveolar. The lobe in one child was a polyalveolar lobe but without retained fetal lung liquid, and one child exhibited conventional lobar emphysema also without retained fetal lung liquid. One polyalveolar lobe caused no neonatal symptoms and was not imaged until the child was 3 months old. No baby with conventional lobar emphysema was shown to have retained fetal lung liquid. There seems to be a correlation between polyalveolar lobe and onset of respiratory symptoms in the first day of life. Retention of fetal lung liquid within the affected lobe was documented only in cases of polyalveolar lobe. PMID- 8414758 TI - Fibromuscular dysplasia with clotted renal artery aneurysm. AB - A six-year-old girl presented with hypertension and headaches. IVP demonstrated a subtle calcification medial to the left kidney and a non-functioning lower pole raising concern for tumor. Color Doppler ultrasound demonstrated normal flow to the upper pole of the left kidney and a severely attenuated avascular lower pole, which was confirmed with selective angiography. Pathology of the surgically resected left kidney showed a calcified large renal artery aneurysm containing thrombus associated with fibromuscular dysplasia. Unique findings in this case were the presence of wall calcification and intraluminal clot in the aneurysm which resulted in nonvisualization of the aneurysm and absent flow to the lower pole. PMID- 8414759 TI - CT and MR in the management of advanced spinal tuberculosis. AB - In regions where tuberculosis is endemic, CT and MR are of limited use in making the initial diagnosis of spinal tuberculosis. Their optimum use is for monitoring the management of cases involving paralysis. CT best evaluates the integrity of posterior bony structures which provide spinal stability and MR best demonstrates the content and extent of the extradural mass that is causing the paralysis. Their use is illustrated by three recent cases. PMID- 8414760 TI - Comparison of CT and MRI in the evaluation of therapeutic response in thoracic Hodgkin disease. AB - Current imaging modalities are accurate in establishing the diagnosis and extent of thoracic Hodgkin disease. After treatment, however, it is extremely difficult to differentiate potential residual active neoplastic disease from scar tissue, or identify early recurrence. We evaluated the contribution of MRI in the assessment of the response to treatment of thoracic Hodgkin disease in the assumption that scar formation would be characterized by low signal intensity in all pulse sequences, whereas active tumor should maintain a degree of high signal intensity on T2-weighted images. In 47 occasions (23 patients) both CT and MRI were able to identify correctly active disease, but had low specificity in confirming remission because of residual tissues masses. High signal intensity on T2-weighted MR images often persisted despite remission, probably because of edema, necrosis, granulation or other factors. MRI was somewhat more specific than CT and may be quite valuable to confirm remission in patients with residual masses that no longer appear hyperintense on T2 after treatment. PMID- 8414761 TI - Cystic retroperitoneal lymphangioma: CT, ultrasound and MR findings. AB - A case of cystic retroperitoneal lymphangioma complicated by hemorrhage is reported in a 7-year-old boy who presented with an abdominal mass. The mass which was partially obstructing the ureter was successfully resected. The imaging findings with emphasis on MR features are described. PMID- 8414762 TI - Gallium scanning in children with fever of unknown origin. AB - Scintigraphy with Gallium 67 is frequently used in the evaluation of children with fever of unknown origin (FUO). Its usefulness in this setting, however, has not been definitely established. We reviewed the clinical records and imaging studies of 30 children with FUO who had Gallium scans. We defined FUO as a febrile illness of greater than two weeks duration which remained undiagnosed after initial clinical, laboratory and radiographic evaluation. 4 of 30 children had positive Gallium scans. Of 25 children with only systemic signs and symptoms in addition to fever, 1 had a positive scan. Of 5 children with more focal complaints, 3 had positive studies: all had localized infections which had remained occult despite imaging with other modalities. We conclude that in most children with FUO, who have only systemic complaints, Gallium scanning is rarely useful. It may be very helpful, however, when there is a suspicion of localized infection, even if other imaging studies are negative. PMID- 8414763 TI - Multiple, symmetrical non-ossifying fibromata without extraskeletal anomalies: report of two related cases. AB - Two patients, mother and daughter, with multiple, symmetrical, non-ossifying fibromata without extraosseous anomalies are reported. We believe that this is a separate syndrome different from multiple non-ossifying fibromata with extraskeletal anomalies (Jaffe-Campanacci syndrome). PMID- 8414764 TI - The Keutel syndrome. Report of a case and review of the literature. AB - A 2-month-old boy presented with cartilage calcification, brachytelephalangism, peripheral pulmonary stenosis, hearing loss, short stature and slight psychomotor delay. This case is similar to the two cases described by Keutel in 1972. Since then, four other cases have been reported; we report the seventh case and discuss the clinical findings and the incidence of the disease. PMID- 8414765 TI - Abdominal ultrasound in Noonan syndrome: a study of 44 patients. AB - Noonan syndrome affects approximately 1 in 1500 live births. Affected individuals may have characteristic phenotypic features some of which are shared with Turner syndrome, although in Noonan syndrome the karyotype is normal, unlike the 45X karyotype of Turner syndrome. Renal anomalies have been described in both syndromes and in Turner syndrome they are both common and frequently severe. The frequency and spectrum of renal anomalies in Noonan syndrome have not been well documented. Upper abdominal ultrasound was performed to establish the frequency of renal anomalies in Noonan syndrome. Forty-four individuals with Noonan syndrome, aged between 9 months and 38 years, were studied. Sixteen scans (36%) were normal and 28 (64%) were abnormal. Five patients (11%) had renal anomalies. Twenty-three patients (53%) had splenomegaly, 6 of these with associated hepatomegaly. One patient had a choledochal cyst and a midgut malrotation. The frequency of renal anomalies in Noonan syndrome is 11%, which is lower than that seen in Turner syndrome. However, splenomegaly with or without hepatomegaly occurs commonly. Choledochal cyst and solitary kidney, previously unreported in Noonan syndrome, are documented. PMID- 8414766 TI - Giant cell reparative granuloma of the base of the skull in a 4-month-old infant- CT findings. AB - An unusual case of giant cell reparative granuloma of the base of the skull of a 4-month-old infant is described. Computerized tomography was useful in defining extent of the lesion and soft tissue abnormalities. Differential diagnosis with other giant cell lesions is discussed. PMID- 8414767 TI - Acromesomelic-spondyloepiphyseal dysplasia associated with congenital optic atrophy: report of a family. AB - We report a family with a unique combination of radiological manifestations of acromesomelic dysplasia and spondyloepiphyseal dysplasia in two members (a man and his daughter) associated with congenital optic atrophy in four generations. The inheritance pattern of this complex anomaly appeared to be autosomal dominant. PMID- 8414768 TI - Lethal short rib syndrome of the Beemer type without polydactyly. AB - A new case of Beemer short-rib dwarfism is reported and the clinical and radiological differences between this and Majewski type are discussed. The clinical variability related to the lack or presence of polydactyly is underlined, together with the importance of prenatal diagnosis. PMID- 8414769 TI - Case report: calculus in the seminal vesicle. AB - Calculus of the seminal vesicle has not been described in children previously. A case is described in which a seminal vesicle calculus was mistakenly thought to lie within the bladder leading to fruitless cystolithotomy. PMID- 8414770 TI - Radiographic patterns of PLH/LIP in HIV positive children. AB - A retrospective study was performed of the radiographic patterns of the lungs in children being followed in the Children's Hospital Aids Program (CHAP). The most consistent finding was a diffuse reticulonodular pattern in the lungs. In addition, lung consolidation, either isolated or in combination with nodules, was noted. There was variation in presentation of the nodules, and prominent linear patterns were frequently encountered. Biopsy specimens of the lungs of these patients demonstrated a correlation between the presence of PLH/LIP and the various radiographic patterns. The lymphoid infiltration of PLH was occasionally observed in a circumferential peri-bronchiolar pattern suggesting an association with the prominent linear patterns on the films. PMID- 8414771 TI - Gas gangrene in a pediatric liver transplant due to infection by Enterobacter cloacae. PMID- 8414772 TI - Ultrasonography in Legg-Calve'-Perthes disease. PMID- 8414773 TI - Salmonella typhi infection in children younger than five years of age. PMID- 8414774 TI - Safety and immunogenicity of Haemophilus influenzae type B-Neisseria meningitidis group B outer membrane protein complex conjugate vaccine mixed in the syringe with diphtheria-tetanus-pertussis vaccine in young Gambian infants. AB - To ensure compliance and to reduce costs it is important, especially in less developed countries, that programs of child immunization should require as few clinic attendances and as few injections as possible. Therefore we have investigated whether a Haemophilus influenzae type b conjugate vaccine could be given safely and effectively with diphtheria-tetanus-pertussis vaccine (DTP). One hundred twenty-six Gambian infants were given both polyribosylribitol phosphate (PRP)-outer membrane protein complex (PedvaxHIB) and DTP on the same day at 8, 12 and 16 weeks of age; 60 were given the vaccines mixed in the syringe and 66 were given the vaccines separately. To minimize the injection volume the dose of PRP OMPC used in both groups was 7.5 micrograms, which is half the usual dose. There were no significant differences in anti-PRP antibody titers between the groups after 1, 2 or 3 doses. The geometric mean titers of antibody for the two groups combined were 0.29 micrograms/ml 1 month after the first dose, 1.03 micrograms/ml after the second dose and 1.11 micrograms/ml after the third dose. Concentrations of antibodies to diphtheria, tetanus and pertussis 1 month after the third dose were not significantly different between the two groups. Systemic side effects were reported with equal frequency in the two groups and were similar to those reported elsewhere for DTP. Tenderness at the injection site was more common where the combined injection (0.75 ml) had been given than where DTP alone (0.5 ml) had been given. The main drawback to the use of these 2 vaccines together is the complexity of the mixing procedure used in this clinical trial. PMID- 8414775 TI - Haemophilus influenzae type b polysaccharide-tetanus protein conjugate vaccine does not depress serologic responses to diphtheria, tetanus or pertussis antigens when coadministered in the same syringe with diphtheria-tetanus-pertussis vaccine at two, four and six months of age. AB - The safety and immunogenicity of a vaccine against Haemophilus influenzae type b consisting of purified polyribosylribitol phosphate conjugated to tetanus toxoid (PRP-T) were evaluated in 277 Chilean infants who were randomly assigned to one of three treatment groups: Group A, PRP-T mixed with diphtheria-tetanus-pertussis (DTP) vaccine in a single syringe and given as a single inoculation in one arm and placebo in the other arm; Group B, PRP-T given in one arm and DTP in the other arm; Group C, DTP given in one arm and placebo in the other. Infants were immunized at 2, 4 and 6 months of age and examined daily for 4 days after each immunization. Serum PRP antibodies; tetanus, diphtheria and pertussis antitoxin; pertussis agglutinins; and antibodies to Bordetella pertussis filamentous hemagglutinin were measured at baseline and 2 months after each dose. PRP-T was well-tolerated. After three doses of PRP-T vaccine 100% of infants attained PRP antibody concentrations > or = 0.15 micrograms/ml and 96 to 99% achieved high anti-PRP concentrations (> or = 1.0 micrograms/ml). The post-third dose anti-PRP geometric mean titer was high (6.94 micrograms/ml) in infants who were given PRP T combined with DTP, although it was somewhat lower than the geometric mean titer of the group who received PRP-T in a separate arm (9.93 micrograms/ml) (P not significant).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414776 TI - Efficacy and safety of dapsone prophylaxis against Pneumocystis carinii pneumonia in human immunodeficiency virus-infected children. AB - Dapsone (4,4'-diaminodiphenylsulfone) is recommended as an alternative agent for prophylaxis against Pneumocystis carinii in children with human immunodeficiency virus infection. We reviewed our experience over the past 100 months with 20 children (age range, 2 months to 13 years) who received dapsone and examined the safety and efficacy of this regimen. Dapsone was taken for an average of 7.33 months/patient or a total of 4410 days by those children in whom safety could be assessed. Three of the 20 patients had an adverse reaction to dapsone. One had mild elevation of blood methemoglobin values (5.6%) and transient elevation of serum transaminases that resolved without discontinuing drug. The other two developed allergic skin rashes which necessitated discontinuation. Efficacy of dapsone in preventing P. carinii pneumonia (PCP) was assessed in 16 children at high risk for developing PCP (defined by CD4 counts or prior PCP infection). These 16 children took dapsone for an average of 6.88 months and a total of 3300 days. Two of the 16 high risk children, one who had had a previous P. carinii pneumonia, developed PCP while taking dapsone. Both had CD4 counts < or = 200 cells/mm3 and were taking dapsone for > or = 12 months before developing PCP. Dapsone is well-tolerated in children and appears to be as effective in preventing PCP in children with human immunodeficiency virus infection as it is in adults. PMID- 8414777 TI - Stimulation of nonspecific immunity to reduce the risk of recurrent infections in children attending day-care centers. The Epicreche Research Group. AB - A randomized, double blind, placebo-controlled clinical trial was performed in 423 children attending day-care centers to assess whether stimulating nonspecific immunity would reduce the incidence of recurrent infections. The drug used for the trial (Imocur) is an extract obtained from eight different species of bacteria. At the end of the total follow-up period (3 months with treatment and 4.5 months without), the risk for > or = 4 episodes of upper respiratory infections was not significantly lower in the treated group than in the placebo group (26.7% vs. 33.8%, relative risk, 0.79; 95% confidence interval, 0.59 to 1.06). In an exploratory analysis limited to the 3-month treatment period, however, we observed a 48% reduction in the risk of presenting > or = 3 episodes of upper respiratory infections: 9.5% vs. 18.3%, respectively, in the treatment group and the placebo group (relative risk, 0.52; 95% confidence interval, 0.31 to 0.86). Similar results were found for the risk of > or = 1 episode of gastroenteritis. We also observed a strong correlation between the drug efficacy and age; this observation is coherent with the underlying pathophysiologic model in which the immune system matures with age. PMID- 8414778 TI - Alpha-2A-interferon for treatment of bronchiolitis caused by respiratory syncytial virus. AB - Alpha-2a-interferon (IFN) has been shown to provide important local defense against some viral infections. In vitro, IFN inhibits the growth of respiratory syncytial virus in bovine monolayer cultures, but respiratory syncytial virus is known to be a poor inducer of IFN in infected infants. When IFN was administered by daily intramuscular injection in a double blind, placebo-controlled study of 22 infants with bronchiolitis caused by respiratory syncytial virus, there was no statistically significant difference in clinical course, duration of oxygen requirement or physical assessment between the treatment and control groups. Likewise viral isolation showed no difference between the groups. Further studies are needed to determine whether higher dosing or aerosol or other alternative route of delivery of IFN might lead to therapeutic benefit. PMID- 8414779 TI - Respiratory syncytial virus infections in hospitalized Canadian children: regional differences in patient populations and management practices. The Pediatric Investigators Collaborative Network on Infections in Canada. AB - Respiratory syncytial virus (RSV) is the most frequent cause of hospitalization for respiratory tract infection during the first 2 years of life. The optimal approach to management remains controversial. During the 1991 to 1992 RSV season RSV-infected children admitted to eight Canadian tertiary care pediatric centers were followed to: (1) assess the morbidity and mortality attributable to RSV infection among hospitalized patients with and without known risk factors for severe disease; and (2) assess regional variation in the management of RSV infection. Of 529 RSV-infected patients 69% (363) had one or more of the risk factors for severe disease and the remaining 31% (166) had none. There were significant differences (P < or = 0.01) between the high and low risk groups, respectively, for: intensive care unit admission (27%, 2%), assisted ventilation (14%, 0.6%), ribavirin therapy (20%, 2%), supplemental oxygen (75%, 34%), antibiotic therapy (69%, 58%) and length of hospital stay > or = 7 days (39%, 6%). Among low risk patients, centers varied significantly (P < or = 0.01) in the use of systemic corticosteroids (from 3 to 69% of patients), supplemental oxygen (13 to 74%), bronchodilators (68 to 93%) and ribavirin (0 to 10%). The observed regional variation in management of hospitalized children with RSV infection has implications for both the costs of hospital care and the conduct of multicenter trials of ribavirin and other therapies for RSV infection. PMID- 8414780 TI - The immune system of human milk: antimicrobial, antiinflammatory and immunomodulating properties. PMID- 8414781 TI - Invasive Aspergillus infections in a pediatric hospital: a ten-year review. PMID- 8414782 TI - Erythromycin-associated hypotension. PMID- 8414783 TI - Pentamidine-induced torsades de pointes. PMID- 8414784 TI - Cardiac arrest after intravenous pentamidine in an infant. PMID- 8414785 TI - Clarithromycin treatment of posttraumatic Mycobacterium fortuitum wound infection. PMID- 8414786 TI - Pertussis and necrotizing enterocolitis in a previously healthy neonate. PMID- 8414787 TI - Recurrent hemorrhagic colitis caused by Escherichia coli O157:H7. PMID- 8414788 TI - Gastrospirillum hominis in a child with chronic gastritis. PMID- 8414789 TI - Painful discoloration of the fingernails in a 15-year-old boy. PMID- 8414790 TI - Hubless syringes and hepatitis B vaccine. PMID- 8414791 TI - Cefixime therapy for otitis media. PMID- 8414792 TI - Vancomycin dosing for neonates. PMID- 8414793 TI - Pet programs in hospitals. PMID- 8414794 TI - Presentation of the Distinguished Physician Award to Stanley A. Plotkin, M.D., by Michael Katz, M.D. PMID- 8414795 TI - Presentation of the Young Investigator Award to Peter J. Hotez, M.D., Ph.D., by Eugene D. Shapiro, M.D. PMID- 8414796 TI - Bayesian forecasting of gentamicin pharmacokinetics in pediatric intensive care unit patients. AB - The predictive performance of a one compartment Bayesian forecasting program was evaluated in pediatric intensive care unit patients with normal renal function. Gentamicin pharmacokinetic parameters were determined in 44 PICU patients (0.8 month to 14 years old) from all available serum concentrations and doses by nonlinear least squares regression. Population pharmacokinetic parameter estimates were established from 27 of the PICU patients. Mean prediction error (ME) and mean absolute error (MAE) for 2 future sets of peak and trough gentamicin serum concentrations with the use of the population parameter estimates with and without feedback were evaluated in the remaining 17 patients. Mean clearance (+/- SD) and volume of distribution for all 44 patients were 0.123 +/- 0.041 liter/hour/kg and 0.424 +/- 0.116 liter/kg, respectively. Bayesian forecasting of the second set of peak and trough concentrations with feedback from the first set of peak and trough concentrations resulted in smaller bias (peak ME, -0.15 mg/liter; trough ME, 0.13 mg/liter) and better accuracy (peak MAE, 0.91 mg/liter; trough MAE, 0.28 mg/liter) compared with the population parameter estimates alone (peak ME, 0.4 mg/liter; trough ME, 0.28 mg/liter; peak MAE, 1.21 mg/liter; trough MAE, 0.57 mg/liter). This study indicates that gentamicin volume of distribution in PICU patients is larger than non-PICU literature values. The Bayesian program, with specific population parameter estimates for PICU patients, provides accurate initial and subsequent predictions of gentamicin serum concentrations. PMID- 8414797 TI - Passive immunization for prevention of rotavirus illness in healthy infants. AB - Infant formula supplemented with bovine antibody (BCIg) to human rotavirus has been reported to prevent or modify rotavirus infection and illness. The purpose of this field trial was to determine the safety and feasibility of passive immunization with bovine antibody for the prevention of rotavirus infection and illness in healthy infants. Sixty-four infants, 31 of whom received BCIg and 33 placebo, participated in the study. Rotavirus infection was detected in 11 (35%) and 14 (42%) infants in the 2 groups, respectively. Symptomatic rotavirus infection was documented in 1 (3%) of the infants receiving BCIg and in 6 (18%) of the infants who received the placebo (P > 0.05). The number of days with diarrhea/1000 days of observation was significantly less in the BCIg group, 4.2, than in the placebo group, 16.8 (P < 0.01). Similarly the number of days with rotavirus-associated diarrhea/1000 days was less in the BCIg group, 1.0, than in the placebo group, 13 (P < 0.01). This study establishes the feasibility of providing passive immunity for the prevention of rotavirus illness in healthy infants. PMID- 8414798 TI - Neonatal tetanus experience at the National Institute of Pediatrics in Mexico City. AB - Clinical charts of 44 neonates admitted to the National Institute of Pediatrics with the diagnosis of neonatal tetanus from 1970 to 1990 were reviewed. All patients had an epidemiologic and clinical findings compatible with neonatal tetanus. Delivery had occurred at the homes of the patients in 89% of the cases and in 11% at clinics. The incubation periods ranged from 2 to 10 days, with a mean of 6.2 days. Cole's periods varied from 1 to 144 hours, with a mean of 21 hours. Spasticity, irritability, refusal to feed, lack of sucking and trismus were present in all cases. Thirty-three patients (70.4%) developed complications, the most frequent being sepsis and bronchopneumonia. The most frequent noninfectious complication was atelectasis, followed by renal failure and electrolytic imbalance. Overall mortality was 25%. It is noteworthy that in the most recent decade (1980 to 1990) mortality was 12.9%, considerably lower than that of the previous decade (1970 to 1980) which was 46.6% (P < 0.008). This decrease was probably a result of the greater availability of mechanical ventilation and the intensive care offered at neonatal services. Mortality was associated with the severity of the disease (P < 0.003) and with the presence of complications (P < 0.025). PMID- 8414799 TI - A comparative evaluation of cefaclor and amoxicillin in the treatment of acute otitis media. AB - In an earlier study of 214 children with acute otitis media (AOM) randomly assigned to a 14-day course of either cefaclor or amoxicillin, 55.7% of cefaclor treated subjects were effusion-free compared with 41.2% of amoxicillin-treated subjects at the end of treatment. The present study was conducted to determine whether, in a 1-year period, subjects treated with cefaclor for each episode would have middle ear effusion for significantly less time than those treated with amoxicillin for each episode. One hundred fifty-seven subjects with AOM were enrolled. No statistically significant differences were found between the cefaclor-treated and amoxicillin-treated groups in number of subjects effusion free immediately after first treatment (47.9% vs. 42.3%, respectively), average percent of time with effusion (22.2% vs. 23.4%, respectively), or in rate of new episodes of AOM (3.05 vs. 3.26, respectively). We conclude that there is no clinically significant advantage to the use of cefaclor rather than amoxicillin as the routine first line drug in the treatment of AOM. PMID- 8414800 TI - Respiratory syncytial virus illnesses in human immunodeficiency virus- and noninfected children. AB - Respiratory syncytial virus (RSV) lower respiratory tract and febrile upper respiratory tract illnesses were prospectively assessed in cohorts of 83 infants born to human immunodeficiency virus (HIV)- and of 48 infants born to non-HIV infected mothers. Of the infants born to HIV-infected mothers, 18 were themselves infected with HIV, 26 were indeterminant and 39 were free from HIV. Ten RSV illnesses occurred in 8 HIV-infected, 2 illnesses in 2 indeterminant and 17 illnesses occurred in 17 non-HIV-infected children. RSV shedding was prolonged in HIV class P2- vs. non-HIV-infected children, at medians of 30 days (range, 1 to 199 days) and 6 days (range, 1 to 21 days), respectively (P = 0.02). Ribavirin and intravenous immunoglobulin failed to eradicate RSV from one child who shed virus for 199 days. Wheezing occurred in 1 of 4 vs. 9 of 10 episodes of lower respiratory tract illness in HIV-infected and non-HIV-infected children, respectively (P = 0.04). No differences were noted in duration of illness, temperature, respiratory rate or oxygen saturation between HIV- and non-HIV infected children. Infection control and public health concerns regarding prolonged shedding of RSV in HIV-infected children must be recognized. PMID- 8414801 TI - Early onset Haemophilus influenzae sepsis in the newborn infant. AB - Neonatal sepsis caused by Haemophilus influenzae is characterized by an early onset syndrome associated with pneumonia, shock and neutropenia. Over a 30-month period 13 infants referred to this hospital had early onset H. influenzae sepsis. Obstetric complications included preterm labor (92%), prolonged rupture of membranes > 12 hours (63%), maternal fever (64%), chorioamnionitis (43%), vaginal discharge (44%) and premature rupture of membranes (15%). All 13 infants were symptomatic at delivery and 7 required immediate intubation. Pneumonia and respiratory distress were the prominent clinical findings. H. influenzae was isolated from infant blood, maternal blood, placenta and genital tract. Isolates were predominantly non-type b, beta-lactamase-negative. A study to determine the prevalence of H. influenzae colonization of the genital tract among women attending clinic at the hospital with the most cases showed a rate of 0.3%. Perinatal risk factors and clinical findings in the infants are similar to disease caused by other organisms associated with early onset sepsis. PMID- 8414802 TI - Role of anaerobic bacteria in liver abscesses in children. AB - Aspirates from pyogenic liver abscesses obtained from 14 children were cultured for aerobic and anaerobic bacteria. Of 29 organisms that were recovered, 17 were anaerobic and 12 were aerobic or facultative. The predominant organisms were Peptostreptococcus spp. (5 isolates). Bacteroides fragilis group (4), Fusobacterium spp. (3) and Staphylococcus aureus (4). Aerobic or facultative bacteria only were recovered in five cases, anaerobic bacteria only in four and mixed aerobic and anaerobic bacteria in five. Anaerobic bacteria were recovered in liver abscesses that were associated with other infection in which these organisms were predominant (i.e. abdominal infection, abscesses). This study highlights the potential importance of anaerobic bacteria in pyogenic liver abscess. PMID- 8414803 TI - Intrauterine parvovirus B19 infection may cause an asymptomatic or recurrent postnatal infection. AB - Although infection with parvovirus B19 (B19) during pregnancy may cause fetal demise, the true incidence of intrauterine infection is unknown. For 19 women with serologically confirmed B19 infections between 4 and 38 weeks of gestation, we performed follow-up examinations of their infants. Serial sonograms of the 19 fetuses showed that none developed hydrops. All 19 women delivered healthy term infants. Cord sera of four infants were tested for IgM to B19 and three were positive. Between 3 and 21 months of age, all 19 infants had normal physical examinations, developmental evaluations and hematocrits; and 16 lacked IgG to B19. One infant who was IgM-positive to B19 at birth was IgM-positive at age 7 months when he also had an IgG titer to B19 of 1:500,000 (mother's concurrent titer, 1:10,000), and had B19 DNA in serum detected by polymerase chain reaction. The other two infants who were IgM-positive at birth were IgM- and IgG-negative by 11 and 16 months of age. These results suggest that intrauterine B19 infection may be frequent and occasionally cause an asymptomatic postnatal infection. PMID- 8414804 TI - Persistent diarrhea in children of developing countries. PMID- 8414805 TI - Streptococcal toxic shock in three Alabama children. PMID- 8414806 TI - Absence of antibiotic-resistant Streptococcus pneumoniae pharyngeal colonization in Cleveland children with sickle cell disease. PMID- 8414807 TI - Bacteremia after exchange transfusion in neonates. PMID- 8414808 TI - Lack of clinical usefulness of a positive latex agglutination test for Neisseria meningitidis/Escherichia coli antigens in the urine. PMID- 8414809 TI - Miliary tuberculosis and symptomatic hypercalcemia. PMID- 8414810 TI - Meningitis with beta-lactam-resistant Streptococcus pneumoniae: the need for early repeat lumbar puncture. PMID- 8414811 TI - Stomatococcus mucilaginosus fatal sepsis in a child with leukemia. PMID- 8414812 TI - Hepatitis B immunization in preterm infants. PMID- 8414813 TI - Viability of Chlamydia trachomatis in eye cosmetics. PMID- 8414814 TI - Human herpesvirus 6 and intussusception. PMID- 8414815 TI - Meningococcal prophylaxis. PMID- 8414816 TI - Serologic tests for Chlamydia pneumoniae. PMID- 8414817 TI - Administration of pertussis and influenza vaccines concurrently. PMID- 8414818 TI - Radiographic sinusitis: illusion or delusion? PMID- 8414819 TI - Controlled trial of oral prednisone in the emergency department treatment of children with acute asthma. AB - BACKGROUND: Recent studies have shown that the use of parenteral corticosteroids in the emergency department decreases the hospitalization rate for patients with acute asthma. We studied the efficacy of oral corticosteroids in the emergency department treatment of moderately ill children with acute asthma. METHODS: Emergency department patients aged 1 through 17 years whose chief complaint was acute asthma were assigned a pulmonary index, based on clinical evaluation. Those with a moderate exacerbation (pulmonary index = 9 through 13) received either 2 mg/kg of oral prednisone or placebo in a randomized, double-blind fashion. Patients in each group were then treated with an identical regimen of frequent aerosolized albuterol, for up to a maximum of 4 hours. RESULTS: Seventy-five patients were assessed. Overall, 11 (31%) of 36 in the prednisone group required hospitalization compared with 19 (49%) of 39 in the placebo group (P = .10). Among the sickest patients (initial pulmonary index > 10), 7 (32%) of 22 prednisone-treated patients required hospitalization compared with 13 (72%) of 18 placebo-treated patients (P < .05). Among patients who had a suboptimal response to initial beta 2-agonist therapy and who therefore would have been hospitalized had treatment been restricted to 2 hours, 9 (45%) of 20 in the prednisone group ultimately required hospitalization when duration of care was extended 2 additional hours compared with 15 (83%) of 18 in the placebo group (P < .05). In addition, prednisone-treated patients had a significantly greater improvement in median pulmonary index (5.0 vs 3.0, P < .001). CONCLUSIONS: These data demonstrate that oral prednisone, within 4 hours of its administration, reduced the need for hospitalization among a subset of children treated in the emergency department for acute asthma. PMID- 8414820 TI - Early treatment of premature infants with recombinant human erythropoietin. AB - OBJECTIVES: The specific objectives of this study were (1) to assess the safety and efficacy of recombinant human erythropoietin (rhEPO) in reducing postnatal hemoglobin decline in premature infants of less than 33 weeks' gestation, and thus reducing the need for transfusion; and (2) to determine the optimal dosage of rhEPO. MATERIALS AND METHODS: Three groups of premature infants of less than 33 weeks' gestation were treated with rhEPO: group 1 (n = 10) received 300 U/kg per week; group 2 (n = 11), 600 U/kg per week; and group 3 (n = 10), 900 U/kg per week. These three groups were compared to a reference group of 20 infants of the same gestational age and birth weight. Treatment started on the 10th day of life and lasted 6 weeks. All infants were given oral iron and vitamin E supplements. RESULTS: Treated infants had significantly higher reticulocyte counts, and the effect was dose dependent (P = .009). Postnatal decline of hemoglobin and hematocrit was lessened in the treated groups; the percent of decrease of hemoglobin and hematocrit was significantly reduced in the treated infants at 35 days of age (P = .0025 and P = .0036, respectively). The need for blood transfusion was also reduced in the rhEPO-treated groups: 19% of treated vs 45% of reference infants received transfusions, and the treated infants received less blood. Serum iron and transferrin saturation percentage dropped significantly during the study and a dose-dependent relationship in treated infants was displayed, suggesting high iron consumption (P = .0008 and P = .006, respectively). No dose effect on hemoglobin level and the need for blood transfusion was found, possibly because of the higher degree of illness severity and iron consumption in groups 2 and 3. No side effects related to rhEPO therapy were observed. CONCLUSIONS: It is concluded that rhEPO therapy is safe in premature babies when given in the three dosages used in this study; in addition, it enhances erythropoiesis and reduces the need for blood transfusions. rhEPO therapy seems more efficient when given in higher dosages; however, illness severity and iron consumption represent major limiting factors. Controlled, randomized studies are warranted to confirm these data and to determine precise modalities and indications of rhEPO therapy in premature infants. PMID- 8414821 TI - The futility of the chest radiograph in the febrile infant without respiratory symptoms. AB - OBJECTIVE: Major pediatric textbooks advocate a chest radiograph as part of the diagnostic evaluation for a sepsis workup for febrile infants less than 3 months old. Very few studies have addressed the value of performing a chest radiograph in this situation. Two studies previously published lack the numbers to statistically justify a conclusion about the need to perform a chest radiograph in the febrile infant. METHODS: Evaluated were 197 febrile infants 3 months old or less with a history, physical examination, chest radiograph, and other laboratory studies to determine the cause of their fever. This group of infants was combined with the group of infants from two similar studies published previously in the literature using cumulative meta-analysis. The combined group resulted in 617 infants. RESULTS: The combined group of infants had 361 infants who had no clinical evidence of pulmonary disease on history or physical examination. All 361 infants had normal chest radiograph. These results gave a 95% confidence interval that the chance of a positive chest radiograph in a patient with no pulmonary symptoms would occur less than 1.02% of the time. CONCLUSIONS: The generally advocated policy of obtaining a chest radiograph as part of the sepsis workup in febrile infants should be discontinued, and chest radiographs should be obtained only in febrile infants who have clinical indications of pulmonary disease. PMID- 8414822 TI - Can seizures be the sole manifestation of meningitis in febrile children? AB - OBJECTIVE: It is frequently taught that lumbar puncture is a mandatory procedure in many or all children who have fever and a seizure, because the convulsion may represent the sole manifestation of bacterial meningitis. We attempted to determine the incidence of this occult manifestation of meningitis. DESIGN: Retrospective case series. SETTING AND PATIENTS: 503 consecutive cases of meningitis in children aged 2 months to 15 years seen at two referral hospitals during a 20-year period. MAIN OUTCOME MEASURES: Signs and symptoms of meningitis in patients having associated seizures. RESULTS: Meningitis was associated with seizures in 115 cases (23%), and 105 of these children were either obtunded or comatose at their first visit with a physician after the seizure. The remaining 10 had relatively normal levels of consciousness and either were believed to have viral meningitis (2) or possessed straightforward indications for lumbar puncture: nuchal rigidity (6), prolonged focal seizure (1), or multiple seizures and a petechial rash (1). No cases of occult bacterial meningitis were found. CONCLUSION: In our review of 503 consecutive children with meningitis, none were noted to have bacterial meningitis manifesting solely as a simple seizure. We suspect that this previously described entity is either extremely rare or nonexistent. Commonly taught decision rules requiring lumbar puncture in children with fever and a seizure appear to be unnecessarily restrictive. PMID- 8414823 TI - Risk factors for acute wheezing in infants and children: viruses, passive smoke, and IgE antibodies to inhalant allergens. AB - OBJECTIVE: To examine the prevalence of viral infection, passive smoke exposure, and IgE antibody to inhaled allergens in infants and children treated for acute wheezing. DESIGN: Case-control study of actively wheezing children who were compared with children without respiratory tract symptoms. SETTING: University of Virginia Pediatric Emergency Room. PATIENTS: Convenience sample of 99 wheezing patients (2 months to 16 years of age) and 57 control patients (6 months to 16 years of age). MEASUREMENTS AND RESULTS: Serum IgE antibody to inhalant allergens, measured by radioallergosorbent test (RAST), was uncommon in wheezing and control patients under age 2. After 2 years of age, the percentage of RAST positive patients increased markedly and was significantly higher in wheezing patients than controls after age 4 (72%, n = 54, and 30%, n = 40, respectively, P < .001). Total IgE levels and nasal eosinophilia were strongly correlated with a positive RAST after age 2. Viral pathogens, predominantly respiratory syncytial virus, were identified in nasal washes from 70% (n = 20) of wheezing patients younger than 2 years of age compared with 20% of controls (n = 10), P < .05. After age 2, viruses, particularly rhinovirus, were isolated in washes from 31% (n = 70) of wheezing patients, 64% of whom were also RAST-positive. Levels of cotinine, a nicotine metabolite, were elevated (> or = 10 ng/mL) in saliva from a large percentage of smoke-exposed, wheezing patients under 2 (74%, n = 19) compared with those over 2 (14%, n = 51), P < .001. Odds ratios for wheezing were significant for virus (8.2, confidence interval [CI] = 1.3 to 5.0), and cotinine (4.7, CI = 1.0 to 21.3) in children under 2, and IgE antibody by RAST (4.5, CI = 2.0 to 10.2), virus (3.7, CI = 1.3 to 10.6), and the combination of IgE antibody and virus (10.8, CI = 1.9 to 59.0) were significant risk factors after age 2. CONCLUSION: Wheezing children younger than 2 years of age had a high rate of viral infection and a low rate of IgE antibody to inhalant allergens. When these children were exposed to passive smoke, salivary cotinine levels were elevated suggesting heavy exposure. After 2 years of age, sensitization to inhaled allergens became increasingly important and viruses remained a significant risk factor for wheezing. These data support recommendations to reduce tobacco smoke exposure at home, especially for young patients, and to consider sensitization to inhaled allergens and allergen avoidance in wheezing children at an early age, particularly after age 2 years. PMID- 8414824 TI - Attitudes and practices regarding analgesia for newborn circumcision. AB - STUDY OBJECTIVE: To determine attitudes regarding routine use of analgesia for newborn circumcision among primary care physicians in southwestern Ontario. DESIGN: Questionnaire survey conducted among all family physicians and pediatricians belonging to the London Academy of Medicine. The majority of pediatric primary care in this region is provided by family physicians. SETTING: Metropolitan center in Southwestern Ontario. RESPONDENTS: A questionnaire was mailed to 279 physicians; 171 (61%) responded. RESULTS: Seventy-four (43%) respondents performed circumcisions. Among physicians performing circumcisions, a minority (17; 24%) used any form of analgesia. The most commonly used analgesic (14; 20%) was oral ethanol. Only 3 (4%) physicians used dorsal penile nerve block. The most common reasons listed for not employing analgesia were lack of familiarity with analgesia use among neonates (56%), lack of familiarity with the technique of dorsal penile block (50%), and concern over adverse effects of analgesic drugs (49%) and regional analgesia (44%). Only a small minority of physicians (7; 12%) responded that they believe that neonates do not feel pain, and 20 (35%) believe that neonates do not remember pain. CONCLUSIONS: Despite evidence that neonates perceive pain and that there is a physiologic stress response to circumcision which can be reduced if analgesia is employed, the vast majority of physicians performing newborn circumcisions either do not employ analgesics or employ analgesics of questionable efficacy. Lack of familiarity with the use of analgesics among neonates and with dorsal penile block in particular are the most common reasons cited for lack of analgesic use. Educational efforts and research into less invasive techniques of analgesia for newborn circumcision are urgently required. PMID- 8414825 TI - Childhood injury prevention counseling in primary care settings: a critical review of the literature. AB - OBJECTIVES: The American Academy of Pediatrics (AAP) believes that health education, through office-based counseling, can contribute to childhood injury prevention. This report presents the results of a critical review of the scientific literature on the effectiveness of primary care-based counseling to prevent childhood unintentional injury. METHODS: A panel selected from the AAP Committee and the AAP Section on Injury and Poison Prevention searched the English-language scientific literature for all articles about childhood unintentional injury prevention counseling. A standardized format was developed to record data on each study. Two members of the panel independently reviewed each article. Articles that were original reports and in which unintentional injury prevention counseling took place in a primary care setting were included. Articles were encoded and analyzed by computer and then grouped by quality of evidence using the US Preventive Services Task Force (USPSTF) method of categorizing results of medical care evaluation. Articles were rated by strength of study design in order to compare studies within each USPSTF group. RESULTS: Twenty articles met the criteria for inclusion. Of these, 18 showed positive effects of injury prevention counseling including five randomized/controlled, 10 non-randomized/controlled, two multiple time series, and one descriptive study. In 15 of the positive studies, physicians performed the counseling. Positive outcomes as measured by increased knowledge, improved behavior, or decreased injury occurrence were reported for both motor vehicle and non-motor vehicle injuries. CONCLUSIONS: The literature review supports the recommendation of the AAP to include injury prevention counseling as part of routine health supervision. This recommendation has implications for health care reimbursement and care content. PMID- 8414826 TI - Crying and motor behavior of six-week-old infants and postpartum maternal mood. AB - OBJECTIVE: To determine whether prepartum and postpartum emotional distress in first-time mothers is associated with crying and fussing behavior and activity level in 6-week-old infants, after eliminating potential biases. DESIGN: Observational study examining predictive and concurrent associations between maternal emotions and infant behaviors. SETTING: General community. PARTICIPANTS: Expectant primiparas attending obstetricians' offices for routine prenatal care were recruited in the third trimester. Mothers and babies were required to be free of medical complications to complete the study. Of 113 mothers who enrolled, complete crying/fussing data were obtained in 88 dyads. Activity data were obtained in a designated subgroup of 50 infants. MEASUREMENTS: Mothers completed a self-report scale of emotional distress, the 28-item General Health Questionnaire, at 34 weeks of gestation and at 6 weeks postpartum. Crying/fussing data were obtained using a previously validated parent diary of infant behavior during the sixth week of life. Motor activity was measured objectively in the home setting with actometers. RESULTS: Third-trimester distress was not related to either infant crying/fussing or activity. Postpartum distress was significantly related to crying/fussing duration and bout frequency (r[88] = .45 and .28, respectively; both P < .01). These relations were not diminished after controlling statistically for background and/or potential mediating variables, nor could they be accounted for by different diary-recording styles in the mothers. Postpartum distress was not, however, related to activity level (r[50] = -.09; not significant). Furthermore, the pattern of maternal distress was associated differentially with crying levels. Distress levels increased from prepartum to postpartum among mothers of infants who met predefined clinical criteria for "colic," while decreasing in the others ("colic" status x period interaction: F(1,86) = 8.2; P < .01). Also, infant crying varied among four groups of mothers who differed according to presence and timing of clinically significant emotional disturbance (one-way analysis of variance, F (3,86) = 9.4; P < .001). Infants of mothers who became significantly distressed postpartum ("reactive") cried more than those in the other groups, even mothers who had been distressed both prepartum and postpartum ("depressed") (3.7 vs 2.7 h/d; P = .05, post hoc Tukey). CONCLUSIONS: Maternal emotional distress and infant crying behavior are associated in the postpartum period independent of reporting or referral biases. Clinically significant levels of crying (or "colic") are differentially associated with different patterns of clinically significant maternal distress. Reported concern about either maternal mood or infant crying behavior should be taken seriously as a possible indicator of a stressed mother infant relationship. PMID- 8414827 TI - Association of the common cold in the first trimester of pregnancy with birth defects. AB - OBJECTIVE: To examine the association between the common cold with or without fever in the first 3 months of pregnancy and birth defects in offspring. DESIGN: A case-control study. SETTING: Data are from the Shanghai Birth Defects Monitoring Program, conducted in 29 hospitals in Shanghai, China from October 1, 1986 to September 30, 1987. SUBJECTS: A total of 986 birth defects cases, 990 frequency-matched live birth controls, and 159 stillbirth controls. RESULTS: Modestly elevated risk of birth defects was identified among women who reported having a cold with or without fever in the first trimester of pregnancy. Notably increased relative risks were observed for anencephalus (odds ratio [OR] = 3.9, 95% confidence interval [CI] = 2.0 to 7.7), spina bifida (OR = 4.1, 95% CI = 1.7 to 9.7), hydrocephalus (OR = 2.3, 95% CI = 1.1 to 5.1), cleft lip (OR = 2.2, 95% CI = 1.4 to 3.4), and undescended testicle (OR = 1.8, 95% CI = 1.0 to 3.0). Our study further found that the overall relative risks were consistent by using two different control groups, suggesting that this association was unlikely to be due to recall or report bias. CONCLUSION: Common cold in the first trimester of pregnancy may be associated with an increased risk of birth defects in offspring. However, these findings should be interpreted cautiously. PMID- 8414828 TI - Increased leukocyte elastase of the tracheal aspirate at birth and neonatal pulmonary emphysema. AB - OBJECTIVE: The authors have previously shown the association of elevated neonatal serum IgM and chorioamnionitis in infants in whom pulmonary emphysema characteristic of Wilson-Mikity syndrome subsequently developed. This paper extends the observation to the measurement of polymorphonuclear leukocyte elastase-alpha 1-proteinase inhibitor complex (PMN elastase-alpha 1-PI) in tracheal aspirates of infants with chronic lung disease. PATIENTS: Tracheal aspirates were obtained from 90 very low birth weight neonates within 24 hours of birth. Serum also was collected within 72 hours of birth, and placentas were examined for signs of inflammation. RESULTS: The mean PMN elastase-alpha 1-PI was significantly elevated (21.8 micrograms/mg albumin) in infants with a pulmonary emphysema syndrome like that designated by Wilson-Mikity compared either with those with bronchopulmonary dysplasia (1.5 micrograms/mg albumin, P < .01) or those with respiratory distress syndrome in whom bronchopulmonary dysplasia did not develop (2.3 micrograms/mg albumin, P < .01). Infants with pulmonary emphysema had a significantly elevated mean serum IgM and a high incidence of chorioamnionitis. CONCLUSIONS: The level of PMN elastase-alpha 1-PI was increased in the tracheal aspirates of newborns in whom pulmonary emphysema developed. Intrauterine inflammation may increase the level of PMN elastase in the fetal respiratory tract. This increase in PMN elastase-alpha 1-PI in fetal lung tissue may cause lung injury in utero, resulting in postnatal pulmonary emphysema consistent with the Wilson-Mikity syndrome following ventilation. PMID- 8414829 TI - Plasma phenylalanine and tyrosine responses to different nutritional conditions (fasting/postprandial) in patients with phenylketonuria: effect of sample timing. AB - OBJECTIVE: To evaluate the adequacy of dietary treatment in patients with phenylketonuria, the monitoring of plasma phenylalanine and tyrosine concentrations is of great importance. The preferable time of blood sampling in relation to the nutritional condition during the day, however, is not known. It was the aim of this study to define guidelines for the timing of blood sampling with a minimal burden for the patient. DESIGN: Plasma concentrations of phenylalanine and tyrosine were measured in nine patients with phenylketonuria who had no clinical evidence of tyrosine deficiency. These values were measured during the day both after a prolonged overnight fast, and before and after breakfast. RESULTS: Phenylalanine showed a small rise during prolonged fasting, while tyrosine decreased slightly. After an individually tailored breakfast, phenylalanine remained stable, while tyrosine showed large fluctuations. CONCLUSION: It is concluded that the patient's nutritional condition (fasting/postprandial) is not important in the evaluation of the phenylalanine intake. To detect a possible tyrosine deficiency, however, a single blood sample is not sufficient and a combination of a preprandial and postprandial blood sample on the same day is advocated. PMID- 8414830 TI - Hepatitis B virus infection in Honolulu students. AB - OBJECTIVES: To determine hepatitis B virus (HBV) carrier and infection rates and HBV epidemiology in Honolulu students. DESIGN: Cross-sectional survey. SETTING: Honolulu public elementary, intermediate, and high schools. PATIENTS: A volunteer sample of 4936 students from 43 Honolulu schools. MAIN OUTCOME MEASURES: HBV carrier rate (presence of hepatitis B surface antigen) and infection rate (presence of either hepatitis B surface antigen, antibody to hepatitis B surface antigen, antibody to hepatitis B core antigen, or any combinations of these) by school grade, ethnicity, and place of birth. RESULTS: Eighty-three (1.68%) students were identified as HBV carriers, and a total of 353 (7.15%) students had serologic evidence of infection. Infection rates increased from elementary school age to high school age, more so in Asian/Pacific Island-born students. The prevalence of infection was higher in Asian/Pacific Island-born students than in those born in the United States. The majority of infected Hawaiian-born students were first-generation Asian/Pacific Island Americans. CONCLUSIONS: Horizontal transmission of HBV occurs in Honolulu school-age children. HBV immunization of all infants in Hawaii, not just those born to carrier mothers, is necessary to reduce HBV transmission in Hawaii. PMID- 8414831 TI - Is there a relationship between dietary fat and stature or growth in children three to five years of age? AB - STUDY OBJECTIVE: To determine whether a moderately reduced fat diet affects the stature or growth of healthy preschool children. DESIGN: Cohort study with mean of 25 months of follow-up. SETTING: Primary care pediatrics practice at a large urban medical center. SUBJECTS: A predominantly Hispanic group of 215 children aged 3 to 4 years at baseline. MEASUREMENTS AND MAIN RESULTS: The children's diet was assessed using four 24-hour recalls and three Willett semiquantitative food frequency questionnaires administered to the children's mothers over a 1-year baseline period. Stature was defined in terms of height, weight, and body mass index at baseline. Growth was defined in terms of change during follow-up in height, weight, and body mass index. Total fat provided a mean of 27.1% of caloric intake in the lowest quintile of intake compared with 38.4% in the highest quintile. There were no differences in stature or growth across quintiles of children defined by consumption of total fat, saturated fat, or cholesterol. These findings were consistent across the two methods of diet assessment. Children who consumed a smaller percentage of total calories from fat consumed significantly less total calories, saturated fat, cholesterol, calcium, and phosphorus, as well as more carbohydrates, iron, thiamine, niacin, vitamin A, and vitamin C. CONCLUSIONS: These data support the safety of a moderately reduced fat diet in healthy preschool children. Maintenance of calcium and phosphorus intake should be part of any program of dietary fat reduction. Substitution of low-fat milk for whole milk, rather than elimination of whole milk, is one such strategy. PMID- 8414832 TI - Assessing the utilization of in-patient facilities in a Canadian pediatric hospital. AB - STUDY OBJECTIVE: To validate the pediatric appropriateness evaluation protocol (P AEP) for use in the Canadian health care system and then to use it to assess the extent of inappropriate utilization in a Canadian tertiary care pediatric facility. METHOD: The P-AEP was applied to a 10% random sample of all general pediatric admissions during 1990 and to a sample of 547 subsequent days of care. The reliability of the P-AEP was assessed using a subsample of 72 admissions and 72 days of care. Validity was tested by comparing the P-AEP judgment on a sample of 50 admissions and 50 days of care with the subjective opinion of panels of three pediatric staff physicians. RESULTS: In the reliability test, there was a high level of agreement between the two independent observers applying the P-AEP. In validity testing, the physicians found a slightly lower rate of inappropriateness relative to the P-AEP, but the validity was good overall. In the main study, 136 of 477 admissions (29%) were found to be inappropriate. Factors associated with inappropriate admission included nonurgent or emergent admission, surgical (versus medical) cases, residence outside the Greater Vancouver area, and admission on Sundays or Mondays. Fifty-five percent of inappropriate admissions were judged necessary but premature, whereas 45% were judged medically unnecessary. Of 547 subsequent days of care, 121 (22%) were found to be medically inappropriate. Inappropriate days of care were associated with girls, Mondays, and patients older than 14 years of age. CONCLUSIONS: The P AEP seems to be a valid and useful instrument for assessing the utilization of pediatric beds in a Canadian health care setting. Using the P-AEP made it possible to identify several service and policy developments which would help improve the efficiency of utilization at the hospital. PMID- 8414833 TI - Fluticasone propionate aqueous nasal spray is safe and effective for children with seasonal allergic rhinitis. AB - INTRODUCTION: Fluticasone propionate aqueous nasal spray, a new topical corticosteroid preparation, is effective when given as 200 micrograms once daily in patients (> 12 years of age) with seasonal allergic rhinitis. STUDY OBJECTIVE: To evaluate the efficacy and safety of fluticasone proprionate aqueous nasal spray in children aged 4 to 11 years with seasonal allergic rhinitis. STUDY DESIGN: Multicenter, randomized, double-blind, placebo-controlled, parallel group. PATIENTS: Two hundred fifty children aged 4 to 11 years with moderate-to severe nasal symptoms, a positive skin test reaction to a late-summer or autumn allergen, a history of seasonal allergic rhinitis, and documentation of an unsatisfactory response to conventional treatment. INTERVENTIONS: Children were randomly assigned to receive fluticasone propionate, either 100 micrograms or 200 micrograms, or placebo, given by intranasal spray once daily in the morning for 14 days. MEASUREMENTS AND RESULTS: Severity of nasal symptoms (obstruction, rhinorrhea, itching, and sneezing) was recorded on visual analog scales by investigators at weekly visits and by patients (or adult guardian) daily in the evening. According to investigator and patient ratings, both fluticasone propionate 100 micrograms/d and 200 micrograms/d lowered total nasal symptom scores when compared with placebo. Both dosages of fluticasone propionate were more effective than placebo on the basis of investigator-rated overall clinical evaluation of efficacy at the end of treatment, with significant improvement (as opposed to moderate or mild improvement, no change or worsening) noted in 21% to 29% of the active-treatment groups vs 9% in the placebo group. There were no significant differences between the two fluticasone propionate dosages in any efficacy measurement. Morning plasma cortisol concentrations and frequency of drug-related adverse events were similar in the fluticasone propionate and placebo groups. CONCLUSION: In children as young as 4 years, 100 micrograms of fluticasone propionate aqueous nasal spray given once daily is as effective as 200 micrograms given once daily, the usual adult dose for the treatment of seasonal allergic rhinitis. Both fluticasone propionate dosages were well tolerated and neither dosage appears to interfere with the hypothalamic-pituitary adrenal axis in children. PMID- 8414834 TI - 'One-touch' electrically operated motor vehicle windows: a potential hazard for children. PMID- 8414835 TI - Treatment of congenital telangiectatic vascular malformations with the pulsed-dye laser (585 nm). PMID- 8414836 TI - Inhaled nitric oxide in the management of a premature newborn with severe respiratory distress and pulmonary hypertension. PMID- 8414837 TI - Advertising in Pediatrics. PMID- 8414838 TI - Toward improving the outcome of pregnancy, 1993: perinatal regionalization revisited. PMID- 8414839 TI - Is neonatal medicine in the United States out of step? PMID- 8414840 TI - Recombinant erythropoietin in anemia of prematurity: five years later. PMID- 8414841 TI - Pediatric surgery: the long road to recognition. PMID- 8414842 TI - American Academy of Pediatrics Task Force on Pediatric AIDS: Addressing concerns of pediatric trainees caring for patients with human immunodeficiency virus infection. PMID- 8414843 TI - American Academy of Pediatrics Task Force on Pediatric AIDS: Adolescents and human immunodeficiency virus infection: the role of the pediatrician in prevention and intervention. PMID- 8414844 TI - American Academy of Pediatrics Committee on Adolescence: Homosexuality and adolescence. PMID- 8414845 TI - International statistics--incomparable. PMID- 8414846 TI - Complication of intranasal midazolam. PMID- 8414847 TI - Hair burning syncope. PMID- 8414848 TI - Different views on adolescent abortion. PMID- 8414849 TI - Different views on adolescent abortion. PMID- 8414850 TI - The principles for family-centered neonatal care. PMID- 8414851 TI - Neonatal hyperbilirubinemia and long-term outcome: another look at the Collaborative Perinatal Project. AB - OBJECTIVE: To examine the association between neonatal bilirubin levels and subsequent neurodevelopmental outcome. DESIGN: Prospective cohort study. SETTING: 12 US medical centers from 1959 (first births) to 1974 (last follow-up). PARTICIPANTS: 41,324 singleton white or black infants with birth weight > or = 2500 g who had neonatal bilirubin measurements recorded and survived at least 1 year. MAIN OUTCOME MEASURES: Wechsler Intelligence Scale for Children Intelligence Quotient (IQ) at age 7 years, blinded neurologic examination at age 7 years, and sensorineural hearing loss at age 8 years. RESULTS: There was no association between IQ and bilirubin. For example, comparing children who had maximum bilirubin levels > or = 342 mumol/L (20 mg/dL) with those who had lower bilirubin levels, adjusted mean IQs were 105.0 and 103.4 in whites (difference + 1.6; 95% confidence interval [CI]: -0.4 to +3.5) and 91.0 and 93.3 in blacks (difference -2.3; 95% CI: -4.8 to +0.2). Abnormal neurologic examination results were reported in 12 of 268 children (4.5%) with bilirubin > or = 342 mumol/L (20 mg/dL) compared with 1249 of 33,004 children (3.8%) with lower levels (relative risk [RR] = 1.2; 95% CI: 0.7 to 2.1). The frequency of abnormal or suspicious neurologic examination results increased in a stepwise fashion with increasing bilirubin level (P < .001), from 4346/29,258 (14.9%) of those with bilirubin levels < 171 mumol/L (10 mg/dL) to 60/268 (22.4%) of those with bilirubin levels. > or = 342 mumol/L (20 mg/dL), apparently due to increasing minor motor abnormalities at higher bilirubin levels. Sensorineural hearing loss was not associated with high bilirubin levels (RR = 1.0; 95% CI: 0.3 to 3.0). CONCLUSIONS: Neonatal bilirubin levels seem to have little effect on IQ, definite neurologic abnormalities, or hearing loss. Higher bilirubin levels are associated with minor motor abnormalities, but the clinical importance of this finding is limited by the weakness of the association, the mild nature of the abnormalities, and the lack of evidence that they are prevented by treatment. PMID- 8414852 TI - Safety of continuous nebulized albuterol for bronchospasm in infants and children. AB - OBJECTIVE: To determine the incidence of cardiotoxicity in infants and children who receive continuous nebulized albuterol (CNA) for bronchospasm. DESIGN: Prospective, case series. SETTING: A university pediatric intensive care and pediatric subacute units. PATIENTS: Nineteen infants and children who received CNA for at least 24 hours. INTERVENTIONS: None. MEASUREMENTS: Creatinine phosphokinase (CK) was measured at the time of admission and then at 12, 24, 48, and 72 hours while the patient received CNA. Isoenzyme CK-MB fractions were measured if CK concentration was > or = 250 IU/L. One electrocardiogram was obtained for each patient during CNA treatment. All patients had continuous cardiac monitoring during continuous nebulization therapy. MAIN RESULTS: Creatinine phosphokinase levels remained within normal limits for 16 patients during CNA treatment. Three patients had elevated CK and in two CK-MB fractions were elevated at one measurement. None of the electrocardiograms showed evidence of ischemia and no arrhythmias were noted during CNA therapy, even in the patients with elevated CK-MB fractions. CONCLUSIONS: Continuous albuterol therapy appears to be safe in our patient population as there was no significant evidence of cardiotoxicity. The significance of the transient elevation of CK-MB without other evidence of cardiotoxicity remains to be determined. PMID- 8414853 TI - After-hours telephone coverage: the application of an area-wide telephone triage and advice system for pediatric practices. AB - BACKGROUND: After-hours telephone calls are a stressful and frustrating aspect of pediatric practice. At the request of private practice pediatricians in Denver, a metropolitan area-wide system was created to manage after-hours pediatric telephone calls and after-hours patient care. This system, the After-Hours Program (AHP), uses specially trained pediatric nurses with standardized protocols to provide after-hours telephone triage and advice for the patients of 92 Denver pediatricians, representing 56 practices. OBJECTIVES: This report describes the AHP, presents data from 4 years' experience with the program, and describes results of our evaluation of the following aspects of the program: subscribing physician satisfaction, parent satisfaction, the accuracy and appropriateness of telephone triage, and program costs. METHODS: After-Hours Program records (including quality assurance data) for all 4 years of operation were retrospectively reviewed, tabulated, and analyzed. The results of two subscribing physician surveys and one parent caller satisfaction survey are presented. A retrospective review of after-hours patient care encounter forms assessed the necessity for after-hours visits triaged by the AHP. An analysis of the total cost of this program to 10 randomly selected subscribing physicians was conducted using current AHP data and a survey of the 10 physicians. RESULTS: In 4 years, 107,938 calls have been successfully managed without an adverse clinical outcome. Minor errors in using protocols occurred in one call out of 1450 after hours calls. After-hours phoen calls necessitated an after-hours patient visit 20% of the time and generated one after-hours hospital admission out of every 88 calls. Just over half of the patients were managed with home care advice only, and 28% were given home care advice after-hours and seen the next day in the primary physician's office. Of all patients directed by the telephone triage nurses to be seen after hours, 78% were determined to have a condition necessitating after-hours care. Data are presented regarding call volumes by time of day, day of week, patient age, and patient's initial complaint. The 6 most common complaints accounted for more than one half of the calls, and 38 complaints accounted for more than 95% of all after-hours calls. Utilization by subscribing physicians is described. Satisfaction among subscribing pediatricians was 100%, and among parents was 96% to 99% on a variety of issues. The total cost to participating Denver pediatricians (which includes revenues "given up" as a result of not seeing patients after hours) ranged from 1% to 12% of their annual net income, depending on a variety of factors. CONCLUSIONS: Large-scale after hours telephone coverage systems can be effective and well-received by patients, parents, and primary physicians. Data presented in this report can assist in planning the training of personnel who provide after-hours telephone advice and triage. Controversies associated with this type of program are discussed. Suggestions are made regarding the direction of future programs and research. PMID- 8414854 TI - Breast-feeding and the risk of life-threatening rotavirus diarrhea: prevention or postponement? AB - PURPOSE: To assess the relationship between breast-feeding and the risk of life threatening rotavirus diarrhea among Bangladeshi infants and children younger than 24 months of age. DESIGN: Case-control study. SETTING: A rural Bangladesh community. PARTICIPANTS: One hundred two cases with clinically severe rotavirus diarrhea detected in a treatment center-based surveillance system during 1985 and 1986, and 2587 controls selected in three surveys of the same community during the same calendar interval. OUTCOMES: Cases and controls were compared for the frequency of antecedent breast-feeding patterns. RESULTS: Compared with other feeding modes, exclusive breast-feeding of infants was associated with significant protection against severe rotavirus diarrhea (relative risk (RR) = 0.10; 95% confidence interval [CI] = 0.03, 0.34). However, during the second year of life, the risk of this outcome was higher in breast-fed than in non-breast-fed children (RR = 2.85; 95% CI = 0.37, 21.71), and no overall protection was associated with breast-feeding during the first 2 years of life (RR = 2.61; 95% CI = 0.62, 11.02). CONCLUSIONS: Although exclusive breast-feeding appeared to protect infants against severe rotavirus diarrhea, breast-feeding per se conferred no overall protection during the first 2 years of life, suggesting that breast-feeding temporarily postponed rather than prevented this outcome. While not detracting from efforts to promote breast-feeding to alleviate the burden of diarrhea due to nonrotaviral enteropathogens, our findings cast doubt on whether such efforts will impact on the problem of severe rotavirus diarrhea. PMID- 8414855 TI - Oxygen saturation and breathing patterns in children. AB - OBJECTIVE: To obtain information on breathing patterns and oxygenation in children. DESIGN: Overnight tape recordings of arterial oxygen saturation (SaO2; Nellcor N200 in beat-to-beat mode), photoplethysmographic waveforms, and breathing movements in 70 healthy children (mean age 8.0 years, range 2 through 16). Analysis of recordings for pauses in breathing movements of > or = 4 seconds (apneic pauses), for episodes in which SaO2 fell to < or = 90% (desaturations) and, only during the state of regular breathing, for baseline SaO2, heart rate, and respiratory rate. RESULTS: Both baseline heart rate and respiratory rate decreased with increasing age (r = -.7 and -.3, respectively, P < .01). Baseline SaO2 was similar to that previously observed in infants (median 99.5%, range 95.8 to 100, 5th centile 96.6%). Every recording showed apneic pauses, with a frequency that did not vary consistently with age (median 7.7/h, range 0.6 to 25.5). One hundred nineteen apneic pauses in 43 recordings lasted for 15 to 19.9 seconds, and 23 lasted for > or = 20 seconds (longest 28.8 seconds). The number of episodic falls in SaO2 to < or = 90% decreased with age (r = -.3, P < .01); such episodes were found in 47% of children aged 2 through 6 years, but in only 13% of those aged 12 through 16 years. The 95th centile for desaturation frequency in the total group was 0.6/h. In six episodes in four patients, SaO2 fell to < or = 80%. CONCLUSIONS: Apneic pauses, some of which can last for more than 20 seconds, are a normal phenomenon in healthy children and adolescents, but only a small minority of apneic pauses affect blood gas homeostasis. Information concerning oxygenation may be more relevant to our understanding of the maturation of respiratory control than the recording of breathing signals alone. PMID- 8414856 TI - Utility of routine laboratory testing for detecting intra-abdominal injury in the pediatric trauma patient. AB - OBJECTIVE: To assess the prevalence of laboratory abnormalities (complete blood cell count, electrolytes, blood urea nitrogen, creatinine, glucose, aspartate aminotransferase, alanine aminotransferase, amylase, lipase, urinalysis [U/A]) and the sensitivity and specificity of the physical examination (PE) and screening laboratory tests for identifying intra-abdominal injury (IAI) in moderately injured pediatric patients. DESIGN, PARTICIPANTS, AND SETTING: Phase I: Retrospective chart review of 285 consecutive level II (moderately injured) trauma patients seen at a children's hospital emergency department/pediatric trauma center. All patients were received directly from the scene and had the following data recorded: mechanism of injury, Glasgow coma score, trauma score, pediatric trauma score, systematically recorded PE findings, laboratory results, and injuries detected during hospitalization. Phase II: To confirm the sensitivity of the PE and U/A found in phase I, the model was applied to 91 additional trauma patients identified by International Classification of Diseases, 9th revision (ICD-9) codes as having IAI. INTERVENTION: None. RESULTS: Phase I: A total of 3939 tests were ordered for the 285 patients entered in phase I. Aspartate aminotransferase and alanine aminotransferase values were obtained in 59% of patients; glucose level was obtained in 78% of patients; complete blood cell count, U/A, and levels of electrolytes, blood urea nitrogen, creatinine, amylase, and lipase were obtained in more than 85% of patients. The overall prevalence of laboratory abnormalities was 5.7%. Fourteen patients (4.8%) were identified who had a total of 23 significant IAIs (9 pancreatic, 6 splenic, 5 renal, 3 hepatic). The PE combined with U/A showing more than five red blood cells per high-power field had a sensitivity of 100%, specificity of 64%, positive predictive value of 13%, and negative predictive value of 100% for the detection of IAI. The presence of laboratory abnormalities suggesting injury did not increase the sensitivity of the model and significantly decreased both specificity and positive predictive value. Phase II: The PE combined with U/A identified an abnormality in 89 (97.8%) of 91 cases (95% confidence interval = 94.8% to 100%). CONCLUSIONS: In the moderately injured pediatric trauma patient, (1) there is a low prevalence of laboratory abnormalities; (2) the PE combined with U/A is a highly sensitive screen for IAI; and (3) in patients with a normal PE of the abdomen and a normal U/A, laboratory testing seldom identifies unsuspected IAI. PMID- 8414857 TI - Human immunodeficiency virus seropositivity in adolescents with syphilis. AB - OBJECTIVE: The purpose of this study was to assess the relationship between syphilis and human immunodeficiency virus (HIV) infection among inner-city, minority group adolescents. METHODS: From August 1989 through June 1990, serum from all positive serologic tests for syphilis, obtained from patients attending a comprehensive adolescent health center in an acquired immunodeficiency syndrome epicenter and its two school-based clinics, were frozen without patient identifiers and were subsequently screened for HIV by enzyme-linked immunosorbent assay with confirmatory Western blot for positives. In addition, a retrospective chart review was performed for all patients with a positive serologic test for syphilis during the study period. RESULTS: Of the 59 specimens with a positive syphilis serologic test, 9 (15.3%) were HIV seropositive. Of the patients with syphilis, 57.4% were black and 42.6% were Hispanic; 16.4% were male (mean age 18.1) and 83.6% were female (mean age 17.8). Only 1 subject (female) was an injection drug user; 4 of the male subjects self-identified as having had sex with other males. Of the subjects, 27.8% had primary, 19.7% had secondary, and 52.5% had latent syphilis at the time of diagnosis. A prior or concurrent sexually transmitted disease was present in 90% of the males and 80% of the females; gonorrhea was the most prevalent sexually transmitted disease in the males (89%) and chlamydia was most prevalent in the females (35%). A history of chancroid and/or herpes was present in 16.4% of the subjects. CONCLUSIONS: It is concluded that the diagnosis of syphilis in an adolescent is a risk factor for HIV infection. All sexually active adolescents should be routinely screened for syphilis, regardless of sexual practices. Those with syphilis should be specifically counseled about their increased risk for HIV infection and the importance of consistent condom use, and they should be referred for formal HIV pretest counseling. PMID- 8414858 TI - Effect of ambient temperature on capillary refill in healthy children. AB - OBJECTIVES: To assess the effect of moderately decreased ambient temperature on capillary refill (CR) time in healthy children, and to measure the reliability of CR measurements between observers. DESIGN: Prospective interventional study with cross-over design. SETTING: Urban pediatric emergency department. PARTICIPANTS: 32 well-hydrated children aged 1 month to 12 years brought to the emergency department for care of minor illness or injury. INTERVENTIONS: Participants were assigned in random order to a 15-minute waiting period in each of two rooms, with and without air-conditioning (cool and warm rooms, respectively). At the end of each waiting period, fingertip CR was measured with a stopwatch, three times by each of one or more three trained observers. RESULTS: Mean CR time was 0.85 +/- 0.45 seconds in the warm room (mean ambient temperature (25.7 degrees C) vs 2.39 +/- 0.76 seconds in the cool room (mean temperature 19.4 degrees C). The mean overall difference in CR time between the two environments was 1.53 seconds (95% confidence interval [CI]: 1.31, 1.75; P < .001); the difference was significant regardless of age or sequence of exposure. 100% of patients were considered to have normal CR (less than 2 seconds) in the warm room, whereas only 31% were considered normal in the cool room. In the 16 patients with CR measured by three observers, interobserver reliability was fair, with an intraclass correlation coefficient of 0.70 (95% CI: 0.56, 0.85), and kappa of 0.54 (95% CI: 0.33, 0.73). CONCLUSIONS: Decreases in ambient temperature within a range found in typical office/emergency department settings may cause significant prolongation of CR time in children with normal circulatory status. There is marked interobserver variability in the measurement of CR even when performed by experienced observers. These findings suggest limitations to the use of CR in the assessment of ill or injured children. PMID- 8414859 TI - Factors affecting left ventricular mass in childhood: the Muscatine Study. AB - OBJECTIVE: To examine the contribution of age, body size, and blood pressure to left ventricular mass (LVM) in childhood and develop a population-based reference of normative LVM data. METHODS: Age, sex, height, weight, and auscultatory systolic and diastolic blood pressures were measured and an echocardiogram was performed to estimate LVM in 904 normal children, aged 6 to 16 years, in Muscatine, IA. Pearson product-moment correlation coefficients were determined to describe the degree of linear association between LVM and age, body size, and blood pressure. Age-sex-, weight-sex-, and height-sex-specific Z scores were determined for LVM, age, weight, height, and blood pressure. Sex-specific LVM prediction equations were derived using weighted-least-squares regression analysis. RESULTS: A strong positive linear association of LVM with age, weight, height, Quetelet index, and systolic and diastolic blood pressure was demonstrated. Z scores for eight different LVM quintile patterns revealed that age, height, weight, and blood pressure each exert an independent influence on LVM in children. Sex-specific predicted M-mode LVM and upper limits of the 90% prediction intervals based on age and height are presented. CONCLUSION: Since age, height, weight, and blood pressure may each exert an independent influence on LVM in children, each factor must be considered when interpreting LVM in childhood. While age, sex, and height are unalterable, both weight and blood pressure can be modified. Thus the pathologic contribution of excess weight and blood pressure ought not be masked by statistical adjustments in reference values for LVM. Sex-specific values of LVM and the 90% and 95% prediction intervals of LVM that do not factor out the effects of obesity or blood pressure are presented. These provide the upper-limit reference values of LVM for the evaluation of children in whom increased LVM is suspected. PMID- 8414860 TI - Safety and efficacy of a topical anesthetic for neonatal circumcision. AB - OBJECTIVE: Circumcision is a common neonatal surgical procedure routinely performed without the use of anesthesia. The purpose of this study was to evaluate the safety and efficacy of topical lidocaine cream as an anesthetic for circumcision. METHODS: Thirty newborns were studied in a randomized, double blind, placebo-controlled study; 15 received a topical 30% lidocaine cream and 15 received the cream base alone. Vital signs were recorded, and preoperative and postoperative serum beta-endorphin and lidocaine concentrations were measured. A videotape of the newborn was used to score behavioral changes. RESULTS: Comparisons of the vital signs precircumcision and postcircumcision showed no differences between the placebo and treatment groups, with the exception of mean systolic blood pressure, which significantly increased in the placebo-treated newborns (P < .05). Serum beta-endorphin concentrations increased postoperatively in 11 of 15 subjects receiving placebo, but decreased or remained unchanged in 10 of 15 subjects receiving lidocaine (P = .03, Fisher's exact test). When stress related behaviors in the precircumcision and post-circumcision periods were compared, the mean increase in their occurrence was greater in the placebo than in the treatment group. There was no significant absorption of lidocaine as measured in the serum. CONCLUSION: Topical application of a 30% lidocaine cream as used in this study may be a safe and efficacious anesthetic for circumcision. PMID- 8414861 TI - Glucocorticoid-responsive hypotension in extremely low birth weight newborns. PMID- 8414862 TI - Hepatitis B immunization in infants of hepatitis B surface antigen-negative mothers. PMID- 8414863 TI - A eutectic mixture of lidocaine and prilocaine for alleviating vaccination pain in infants. PMID- 8414864 TI - Fluoxetine hydrochloride (Prozac) toxicity in a neonate. AB - A case of fluoxetine toxicity in a newborn of 38 weeks' gestation has been presented. The total drug concentration in cord blood was 80 ng/mL. The fluoxetine level, 26 ng/mL, is below the adult therapeutic level; the norfluoxetine cord blood level, 54 ng/mL, is at the adult therapeutic level. At 96 hours the fluoxetine level was not measurable and the norfluoxetine level was 55 ng/mL. The parent compound is fluoxetine, which is metabolized in the liver to norfluoxetine. The half-life of fluoxetine is 2 to 3 days, and that of norfluoxetine is 7 to 9 days. Interestingly, in our patient the fluoxetine was absent in the blood at 4 days, but norfluoxetine was present. The most common side effects of Prozac in adult patients involve primarily the central nervous system and include nervousness, tremor, jitteriness, and occasionally seizures. Central nervous system symptoms were most prominent in this newborn. He also had an increased heart rate. Cardiovascular side effects are less prominent in adults who are taking Prozac. The neonate in this case was asymptomatic at 96 hours of age, indicating that the parent compound, fluoxetine, may be the active part of the drug and side effects may be caused by the parent compound. PMID- 8414865 TI - Capillary refill: is it a useful clinical sign? PMID- 8414866 TI - Parent dissatisfaction with neonatal intensive care unit care and suggestions for improvements. PMID- 8414867 TI - American Academy of Pediatrics Committee on Careers and Opportunities: Pediatric workforce statement. PMID- 8414868 TI - American Academy of Pediatrics Committee on Psychosocial Aspects of Child and Family Health: The pediatrician and the "new morbidity". PMID- 8414869 TI - American Academy of Pediatrics Committee on Child Abuse and Neglect and Committee on Community Health Services: Investigation and review of unexpected infant and child deaths. PMID- 8414870 TI - Questions about electrographic seizures in newborns. PMID- 8414871 TI - Shunts in patients with respiratory distress syndrome. PMID- 8414872 TI - Prevention of fetal alcohol syndrome. PMID- 8414873 TI - Prevention of fetal alcohol syndrome. PMID- 8414874 TI - Hearing loss not associated with DTP vaccination or fever. PMID- 8414875 TI - Arterial catheters and retinopathy of prematurity risk: need for a multicenter controlled trial. PMID- 8414876 TI - Unlisted ingredients. PMID- 8414877 TI - Focal visual attention and pattern discrimination. AB - Pattern discrimination in the presence of distractor patterns is improved when the stimulus display is preceded by a precue designating the location of the target pattern. Experiments were conducted to determine how big an improvement the precue produced. The specific question of whether the observer is able to process selectively the stimulus pattern in the cued location of the display and ignore the patterns of the noncued locations was addressed. In order to study this, reaction time for pattern discrimination on a blank background (no distractors) was compared with the reaction time when the observer performed the same discrimination task in the presence of distractors and a precue had indicated the location of the stimulus pattern to be discriminated. The results showed that these two reaction times were equal if the cue preceded the stimulus patterns at intervals which were longer than some minimum time. Hence, stimuli outside the 'aperture' of focal attention can be ignored. These results could not be attributed to eye movements, because the longest duration of the whole sequence of precue and stimulus patterns was only 200 ms. PMID- 8414878 TI - Voluntary and stimulus-induced attention detected as motion sensation. AB - Attention may be drawn passively to a visually salient object. We may also actively direct attention to an object of interest. Do the two kinds of attention, passive and active, interact and jointly influence visual information processing at some neural level? What happens if the passive and active attentions come into conflict? These questions were addressed with the aid of a novel psychophysical technique which reveals an attentional gradient as a sensation of motion in a line which is presented instantaneously. The subjects were asked to direct attention with voluntary effort: to the side opposite to a stimulus change, to an object with a predetermined colour, and to an object moving smoothly. In every case the same motion sensation was induced in the line from the attended side to the unattended side. This voluntary attention, however, can easily and quickly be distracted by a change in the periphery, though it can be regained within a period of 200 to 500 ms. The results suggest that the line motion can be induced in voluntary (top-down) as well as stimulus-driven (bottom up) situations, thus indicating the truly attentional nature of the effect, rather than it being some kind of retinotopic sensory artifact or response bias. The results also suggest that these two kinds of attention have facilitatory effects acting together on a relatively early stage of visual information processing. PMID- 8414879 TI - Up-down asymmetry in vertical induced motion. AB - Induced motion (IM) is the illusory movement of an object in the direction opposite to the real motion of adjacent detail. One theory of IM suggests that it results, in part, from suppression of optokinetic nystagmus (OKN) by fixational (smooth-pursuit) effort. In several studies an asymmetry in human vertical OKN has been reported, with upward optokinetic stimulation eliciting higher OKN gain than downward motion. This provides a test of the nystagmus-suppression theory of IM. If suppression of OKN contributes significantly to IM, upward inducing stimuli should result in a greater magnitude of the illusion than should downward stimulus motion. Additionally, the asymmetry of vertical OKN should become more pronounced at higher stimulus velocities. Therefore, the asymmetry of vertical IM should be greater at higher inducing-stimulus velocities. Twelve subjects viewed a large, random-dot stimulus, which moved either upward or downward at a velocity of 10, 40, or 70 deg s-1. Subjects fixated a horizontally moving laser spot and adjusted a rod to match the apparent slope of the motion path of the spot. IM magnitude was derived from these measures. Mean IM velocity was significantly higher with upward than with downward stimulation, and the difference was maximal at velocities of 40 and 70 deg s-1. The results are discussed within the context of the nystagmus-suppression theory and other theories of IM. PMID- 8414880 TI - Apparent relative size and depth of moving objects. AB - The effects of the relative velocities of moving objects on their apparent relative size and depth were investigated with the aid of square patterns generated on a CRT display by a microcomputer. The observer matched the apparent sizes of squares, arranged in two or more rows that moved with different velocities, and made judgments of the apparent relative depth of the rows. In many conditions, squares moving more slowly were perceived as larger in size than those moving faster, regardless of the kind of depth responses, but in some conditions which contained strong depth cues the size responses seemed to be affected by perceived depth. The size-change effect of moving objects is discussed from the viewpoint of size-depth relationship. PMID- 8414881 TI - Estimates of time to contact based on changing size and changing target vergence. AB - Apparent constant-velocity approaches of an outline circle were generated by changing the size and/or the lateral separation of dichoptically presented circles at a physically constant distance from subjects. The lateral separation of the circles defines target vergence which corresponds proximally to combinations of ocular vergence and (absolute) retinal disparity. In the first experiment, estimates of time to contact were found to depend both on changing size and on changing target vergence; in the case of conflicting information, changing size turned out to be the more powerful stimulus. In the second experiment, the size of the stimulus object was varied. The relatively stronger influence of changing size on estimates of time to contact was again found for the larger stimulus objects, but for the smaller stimulus objects changing target vergence became more powerful. The results of both experiments show that estimates of time to contact are not uniquely determined by changing size, as seems to be implied by some tests of the so-called "tau hypothesis', but that they are based on multiple sources of information that have situationally specific effects on the estimates. PMID- 8414882 TI - Effects of element type and spatial grouping on symmetry detection. AB - The influence of local and global attributes of symmetric patterns on the perceptual salience of symmetry was investigated. After tachistoscopic viewing, subjects discriminated between symmetric and either random patterns (experiment 1) or their perturbed counterparts (experiment 2) created by replacing one third of the mirror element-pairs of symmetric stimuli with 'random' elements. In general, it was found that perceptibility of symmetry, measured by response time and detection accuracy, was not influenced in a consistent way by type of pattern element (dots or line segments oriented vertically, horizontally, obliquely, or in all three orientations about the symmetry axis). Nor did axis orientation (vertical, horizontal, oblique), advance knowledge of axis orientation, practice effects, or subject sophistication differentially affect detection. A highly salient global percept of symmetry emerged, on the other hand, when elements were clustered together within a pattern, or grouped in symmetric pairs along a single symmetry axis or two orthogonal axes. Results suggest that mirror symmetry is detected preattentively, presumably by some kind of integral code which emerges from the interaction between display elements and the way they are organized spatially. It is proposed that symmetry is coded and signalled by the same spatial grouping processes as those responsible for construction of the full primal sketch. PMID- 8414883 TI - Parallelism and the perception of illusory contours. AB - The role of symmetry in the perception of illusory contours has been a subject of controversy ever since Kanizsa proposed his theory of illusory contours based on Gestalt principles. Today it is widely agreed that illusory contours do not necessarily occur more readily with inducers that can be 'amodally' completed to symmetrical objects than with inducers that cannot. But the question of whether symmetrical inducers produce weaker illusory contours than do unsymmetrical ones is still controversial. A novel determinant of illusory contour strength, parallelism, is proposed. Experiments are reported which indicate that illusory contours induced by 'blobs' which have boundaries that are nearby and parallel to the illusory contour are weaker than illusory contours induced by blobs that do not have this property. It is suggested that the display that has been most widely used by researchers to support their claims for a weakening of illusory contours with symmetrical inducers is weak primarily because of parallelism. PMID- 8414884 TI - Toward a computational model of constraint-driven exploration and haptic object identification. AB - A conceptual model of the human haptic system in relation to object identification is presented. The model encompasses major architectural elements including representations of haptically accessible object properties and exploratory procedures (EPs)--dedicated movement patterns that are specialized to extract particular properties. These architectural units are related in processing-specific ways. Properties are associated with exploratory procedures in keeping with the extent to which a given procedure delivers information about a given property. The EPs are associated with one another in keeping with their compatibility, as determined by parameters of motor execution and interactions with the object and the workspace. The resulting architecture is treated as a system of constraints which guide the exploration of an object during the course of identification. The selection of the next step in a sequence of exploration requires that constraints be optimally satisfied. A network approach to constraint satisfaction is implemented and shown to account for a number of previous empirical results concerning the time course of exploration, object classification speed, and incidental learning about object properties. This system has potential applications for robotic haptic exploration. PMID- 8414885 TI - Discrimination of the shape of the masked inducing figure precedes perception of the illusory triangle. AB - Using the Kanizsa triangle in a free-inspection paradigm, Gellatly has shown that subjects report seeing an illusory triangle while apparently not phenomenally aware of the inducing areas. It is argued that Gellatly's procedure may induce response processes which camouflage early sensory processes. By forcing subjects to choose between two response alternatives, it was shown that when they could just perceive the illusory triangle, they could also correctly discriminate the shape of the inducing elements. Under the free-inspection procedure, the duration threshold for seeing the illusory triangle was about 9 ms while the threshold for discriminating the shape of the inducing elements was about 2 ms. A static representation of the dynamic procedure used by Gellatly has been derived, which shows a novel effect of contour completion across boundaries resulting in a ghostly white triangle. PMID- 8414886 TI - Tempo sensitivity in auditory sequences: evidence for a multiple-look model. AB - Differential thresholds for tempi (with interonset intervals ranging from 100 to 1,500 msec) were measured using an adaptive 2IFC paradigm for several types of auditory sequences. In Experiment 1, the number of intervals in an isochronous sequence was varied to compare the sensitivity for single intervals with that for sequences of two to six intervals. Mean relative just noticeable differences (JNDs) decreased as the number of intervals increased (single intervals = 6%, two intervals = 4%, four intervals = 3.2%, six intervals = 3%) and were optimal at intermediate tempi for both sequences and single intervals (as low as 1.5% in the range between 300 and 800 msec). In Experiment 2, the sensitivity for different types of irregular sequences was studied. Globally, JNDs for irregular sequences were of an intermediate level between that observed for single intervals and that observed for regular sequences. However, the closer a sequence was to regularity, the lower its relative JND. Experiment 3 demonstrated that musicians were more sensitive than nonmusicians to changes in tempo, and this was true for single intervals and for regular and irregular sequences, demonstrating the role of training on these abilities. The results are discussed in terms of possible underlying mechanisms, in particular those providing a mental representation of the mean and dispersion of successive interval durations. PMID- 8414887 TI - Phonotactic knowledge of word boundaries and its use in infant speech perception. AB - The development of a lexicon critically depends on the infant's ability to identify wordlike units in the auditory speech input. The present study investigated at what age infants become sensitive to language-specific phonotactic features that signal word boundaries and to what extent they are able to use this knowledge to segment speech input. Experiment 1 showed that infants at the age of 9 months were sensitive to the phonotactic structure of word boundaries when word-like units were presented in isolation. Experiments 2 to 5 demonstrated that this sensitivity was present even when critical items were presented in context, although only under certain conditions. Preferences for legal over illegal word boundary clusters were found when critical items were embedded in two identical syllables, keeping language processing requirements and attentional requirements low. Experiment 6 replicated the findings of Experiment 1. Experiment 7 was a low-pass-filtered version of Experiment 6 that left the prosody of the stimulus items intact while removing most of the distinctive phonotactic cues. As expected, no listening preference for legal over illegal word boundary clusters was found in this experiment. This clearly suggests that the preferential patterns observed can be attributed to the infants' sensitivity to phonotactic constraints on word boundaries in a given language and not to suprasegmental cues. PMID- 8414888 TI - Variability of olfactory threshold and its role in assessment of aging. AB - Olfactory thresholds of elderly persons (over 65 years) average one to two orders of magnitude higher than those of young adults (under 30 years). Past studies reveal enormous spreads (typically about three orders of magnitude) of individual thresholds within each age group and extensive overlap between the two groups- enough to question how typically decline in sensitivity characterizes the individual aged person. The present study shows that much of the observed overlap is misleading, because the brief threshold tests usually administered tend to exaggerate individual differences. A more representative assessment of an individual's threshold (for 1-butanol) was achieved by averaging the thresholds from two to eight separate short tests, spread over 4 days. The spread of each group's thresholds (12 young and 12 elderly subjects) narrowed strikingly as the number of tests averaged increased from one to four; further tests accomplished no additional narrowing of spread. Based on a single test, thresholds of young and elderly overlapped in the usual way; but based on four or more tests, thresholds of young and elderly overlapped little or not at all. The outcome (1) argues that decline in smell sensitivity seems to be, after all, a common feature of aging, and (2) sheds light on the sources of variability of sensory thresholds. PMID- 8414889 TI - The role of timbre in the segregation of simultaneous voices with intersecting F0 contours. AB - When the fundamental frequency (F0) contours of two speakers' voices intersect, the listener is presented with a problem. The listener must decide which of the F0 contours emerging from the intersection is a continuation of which contour entering the intersection: have the F0 contours crossed or merely approached and parted? In the present experiment, subjects listened to two simultaneous diphthong-like sounds with F0 contours that either approached and diverged or crossed over. The task was to report whether the pitches "crossed" or "bounced" away from each other. Despite the changing timbres of the two sounds, the subjects were able to discriminate crossing and bouncing F0s, provided that the timbres of the vowels differed at the moment when their F0s were the same. When the timbres were the same, the subjects could not make the discrimination and tended to hear a bouncing percept. These results are consistent with the idea that listeners use continuity of timbre rather than continuity of F0 movement to disambiguate F0 intersections. PMID- 8414890 TI - Developmental trends in the interaction between auditory and linguistic processing. AB - The developmental course of multidimensional speech processing was examined in 80 children between 3 and 6 years of age and in 60 adults between 20 and 86 years of age. Processing interactions were assessed with a speeded classification task (Garner, 1974a), which required the subjects to attend selectively to the voice dimension while ignoring the linguistic dimension, and vice versa. The children and adults exhibited both similarities and differences in the patterns of processing dependencies. For all ages, performance for each dimension was slower in the presence of variation in the irrelevant dimension; irrelevant variation in the voice dimension disrupted performance more than irrelevant variation in the linguistic dimension. Trends in the degree of interference, on the other hand, showed significant differences between dimensions as a function of age. Whereas the degree of interference for the voice-dimension-relevant did not show significant age-related change, the degree of interference for the word-dimension relevant declined significantly with age in a linear as well as a quadratic manner. A major age-related change in the relation between dimensions was that word processing, relative to voice-gender processing, required significantly more time in the children than in the adults. Overall, the developmental course characterizing multidimensional speech processing evidenced more pronounced change when the linguistic dimension, rather than the voice dimension, was relevant. PMID- 8414891 TI - Detection of three-dimensional surfaces from optic flow: the effects of noise. AB - Previous research (Andersen, 1989) has suggested that the recovery of 3-D shape from nonsmooth optic flow (motion transparency) can be performed by segregating surfaces according to the distributions of velocities present in the flow field. Five experiments were conducted to examine this hypothesis in a surface detection paradigm and to determine the limitations of human observers to detect 3-D surfaces in the presence of noise. Two display types were examined: a flow field that simulated a surface corrugated in depth and a flow field that simulated a random volume. In addition, two types of noise were examined: a distribution of noise velocities that overlapped or did not overlap the velocity distribution that defined the surface. Corrugation frequency and surface density were also examined. Detection performance increased with decreasing corrugation frequency, decreasing noise density, and decreasing surface density. Overall, the subjects demonstrated remarkable tolerance to the presence of noise and, for some conditions, could discriminate surface from random conditions when noise density was twice the surface density. Discrimination accuracy was greater for the nonoverlapping than for the overlapping noise, providing support for an analysis based on the distribution of velocities. PMID- 8414892 TI - Estimating local shape from shading in the presence of global shading. AB - In theory, global shading may help with the estimation of local surface structure from shading (e.g., in specifying the illuminant direction). Empirically, we do not know whether human observers combine the information given by the local and global shading to estimate local shape. Observers had to indicate the orientation of a local elongated perturbation with or without global shading information provided by a background surface. Our psychophysical results show the following: 1. Observers do not estimate the orientation of the local perturbation more accurately with global shading information than they do in the absence of such information. 2. Responses depend dramatically on the inclination between the illuminant direction and the viewing direction. For an inclination of 20 degrees, observers indicate more or less the orientation of the local ridge; however, for an inclination of 40 degrees, they indicate either the direction of the illuminant or an orientation close to the shadow edge of the perturbation. Most subjects show some combination of these behaviors. This behavior is not altered by global shading information. We conclude that in our paradigm, global shading information does not aid the estimation of local shape. PMID- 8414893 TI - Both perceptual and conceptual factors influence taste-odor and taste-taste interactions. AB - Observers are often asked to make intensity judgments for a sensory attribute of a stimulus that is embedded in a background of "irrelevant" stimulus dimensions. Under some circumstances, these background dimensions of the stimulus can influence intensity judgments for the target attribute. For example, judgments of sweetness can be influenced by the other taste or odor qualities of a solution (Frank & Byram, 1988; Kamen et al., 1961). Experiments 1 and 2 assessed the influence of stimulus context, instructional set, and reference stimuli on cross quality interactions in mixtures of chemosensory stimuli. Experiment 1 demonstrated that odor-induced changes in sweetness judgments were dramatically influenced when subjects rated multiple attributes of the stimulus as compared with when they judged sweetness alone. Several odorants enhanced sweetness when sweetness alone was judged, while sweetness was suppressed for these same stimuli when total-intensity ratings were broken down into ratings for the sweetness, saltiness, sourness, bitterness, and fruitiness of each solution. Experiment 2 demonstrated a similar pattern of results when bitterness was the target taste. In addition, Experiment 2 showed that the instructional effects applied to both taste-odor and taste-taste mixtures. It was concluded that the taste enhancement and suppression observed for taste-odor and taste-taste mixtures are influenced by (1) instructional sets which influence subjects' concepts of attribute categories, and (2) the perceptual similarities among the quality dimensions of the stimulus. PMID- 8414894 TI - Prismatic displacement of vision induces transient changes in the timing of eye hand coordination. AB - Eye-hand coordination was investigated during a task of finger pointing toward visual targets viewed through wedge prisms. Hand and eye latencies and movement times were identical during the control condition and at the end of prism exposure. A temporal reorganization of eye and hand movements was observed during the course of adaptation. During the earlier stage of prism exposure, the time gap between the end of the eye saccade and the onset of hand movement was increased from a control time of 23 to 68 msec. This suggests that a time consuming process occurred during the early prism-exposure period. The evolution of this time gap was correlated with the evolution of pointing errors during the early stage of prism exposure, in such a way that both measures increased at the onset of prism exposure and decreased almost back to control values within about 10 trials. However, spatial error was not entirely corrected, even late in prism exposure when the temporal organization of eye and hand had returned to baseline. These data suggest that two different adaptive mechanisms were at work: a rather short-term mechanism, involved in normal coordination of spatially aligned eye and hand systems, and a long-term mechanism, responsible for remapping spatially misaligned systems. The former mechanism can be strategically employed to quickly optimize accuracy in a situation involving misalignment, but completely adaptive behavior must await the slower-acting latter mechanism to achieve long-term spatial alignment. PMID- 8414895 TI - A comparison of thresholds for 1/3-octave filtered clicks and noise bursts in infants and adults. AB - Behavioral thresholds of 6-month-old infants and adults were determined for 1/3 octave filtered clicks and 300-msec noise bursts with center frequencies ranging from .5 to 8 kHz. For noise bursts, differences between infant and adult thresholds were largest at low frequencies and smallest at 8 kHz. For clicks, infants' thresholds were most like adults' at 4 kHz, and age differences increased at both lower and higher frequencies. Differences between click and noise thresholds were significantly larger for infants than for adults at .5, 1, and 8 kHz, but not at 2 and 4 kHz. These results suggest that improvements in threshold for long-duration stimuli during infancy may not be accompanied by comparable changes in threshold at short durations. The delayed development of sensitivity to low- and high-frequency clicks appears consistent with maturational trends recently described for the auditory brainstem response. PMID- 8414896 TI - The effect of tempo and tone duration on rhythm discrimination. AB - Rhythm constancy was investigated in two experiments. In Experiment 1, the first rhythm was presented at one tempo, the second rhythm was presented at a different tempo, and subjects judged whether the relative timing structures were identical (i.e., was the first rhythm merely sped up or slowed down to generate the second rhythm?). For the nonmetric rhythms used here, subjects perceived the rhythm in terms of the figural grouping of the tones, and rhythm constancy broke down between slower and faster tempos. In Experiment 2, the first rhythm was presented in tones of one duration; the second rhythm was presented in tones of a different duration; and subjects judged whether the timing structures of the tone onsets were identical (the two rhythms were presented at the same tempo). These results indicated a high degree of constancy; subjects found it easy to discriminate the timing structures. These results confirm that the onset timing is critical to rhythm perception and suggest that rhythm perception at slower rates (2 elements/sec) differs from rhythm perception at faster rates (3-4 elements/sec). PMID- 8414897 TI - Duration discrimination of empty and filled intervals marked by auditory and visual signals. AB - Experiments 1 and 2 compared, with a single-stimulus procedure, the discrimination of filled and empty intervals in both auditory and visual modalities. In Experiment 1, in which intervals were about 250 msec, the discrimination was superior with empty intervals in both modalities. In Experiment 2, with intervals lasting about 50 msec, empty intervals showed superior performance with visual signals only. In Experiment 3, for the auditory modality at 250 msec, the discrimination was easier with empty intervals than with filled intervals with both the forced-choice (FC) and the single stimulus (SS) modes of presentation, and the discrimination was easier with the FC than with the SS method. Experiment 4, however, showed that at 50 and 250 msec, with a FC-adaptive procedure, there were no differences between filled and empty intervals in the auditory mode; the differences observed with the visual mode in Experiments 1 and 2 remained significant. Finally, Experiment 5 compared differential thresholds for four marker-type conditions, filled and empty intervals in the auditory and visual modes, for durations ranging from .125 to 4 sec. The results showed (1) that the differential threshold differences among marker types are important for short durations but decrease with longer durations, and (2) that a generalized Weber's law generally holds for these conditions. The results as a whole are discussed in terms of timing mechanisms. PMID- 8414898 TI - Parameters affecting gap detection in the rat. AB - The present research used a startle amplitude reduction paradigm to investigate the ability of the rat's auditory system to track rapidly changing acoustic transients. Specifically examined was the ability of brief gaps in otherwise continuous noise to reduce the amplitude of a subsequently elicited acoustic startle reflex. The duration of the gap, time between gap offset and startle elicitation (the interstimulus interval or ISI), and rise-fall characteristics of the gap were systematically varied. Consistent with previous research, gaps reliably reduced startle amplitude. Gaps 2 msec long were reliably detected, and a 50-msec ISI resulted in the greatest amplitude reduction. Gaps presented at short ISIs produced amplitude reduction that followed a different time course than did gaps presented at longer ISIs. These results may reflect differences in the length of time available for the processing of the stimulus and may involve two different processes. PMID- 8414899 TI - Visual influences on auditory pluck and bow judgments. AB - In the McGurk effect, visual information specifying a speaker's articulatory movements can influence auditory judgments of speech. In the present study, we attempted to find an analogue of the McGurk effect by using nonspeech stimuli- the discrepant audiovisual tokens of plucks and bows on a cello. The results of an initial experiment revealed that subjects' auditory judgments were influenced significantly by the visual pluck and bow stimuli. However, a second experiment in which speech syllables were used demonstrated that the visual influence on consonants was significantly greater than the visual influence observed for pluck bow stimuli. This result could be interpreted to suggest that the nonspeech visual influence was not a true McGurk effect. In a third experiment, visual stimuli consisting of the words pluck and bow were found to have no influence over auditory pluck and bow judgments. This result could suggest that the nonspeech effects found in Experiment 1 were based on the audio and visual information's having an ostensive lawful relation to the specified event. These results are discussed in terms of motor-theory, ecological, and FLMP approaches to speech perception. PMID- 8414900 TI - Binocular rivalry of equiluminant targets. PMID- 8414901 TI - A two-step model of secretion control in neuroendocrine cells. AB - Recent experiments on a variety of neuroendocrine cells indicate that intense stimuli readily depress the secretory response. The most likely explanation for this depression is that a pool of release-ready granules is depleted. We present a two-step model of secretion that allows one to simulate the dynamics of such a pool for different time courses of free intracellular Ca concentration [Ca2+]i. We derive rate constants of the model from two types of experiment and find that, for the simplest type of model, not only the rate of consumption (exocytosis) but also the rate of vesicle supply to the pool of release-ready granules must be made Ca-dependent. Given these functional dependences a variety of results from the literature can be simulated. In particular, the model predicts the occurrence of secretory depression and augmentation under appropriate conditions. PMID- 8414902 TI - Comparison of DNA synthesis in white and brown adipose tissue in rats with ventromedial hypothalamic lesions. AB - Ventromedial hypothalamic (VMH) lesions cause excessive fat accumulation in white adipose tissue (WAT), and brown adipose tissue (BAT); however, little information is available on whether or not cell proliferation occurs in WAT and BAT after VMH lesioning. In this study, we determined the DNA content and thymidine incorporation in unilateral parametrial WAT and interscapular BAT 0, 1, 3, and 7 days after VMH lesioning, and examined the mechanism of increased DNA content in WAT. In rats with VMH lesions, the weight of WAT and BAT had increased significantly at 7 days, and the DNA content and thymidine incorporation of WAT had increased significantly at 3 days and continued to increase for up to 7 days, while those of BAT did not increase for as long as 7 days after VMH lesioning. Restricted food intake according to the pair-feeding method partially inhibited the increased DNA content in WAT. The increased DNA content in WAT was mostly restored but not completely by the administration of anti-insulin antibody, and by administration of propranolol, a beta-adrenergic blocker. The results demonstrated that VMH lesions induced DNA synthesis in WAT early after VMH lesioning, but did not induce DNA synthesis in BAT, and suggested that either hyperinsulinemia or a beta-adrenergic receptor mechanism or both may be responsible for the increased DNA content in WAT. PMID- 8414903 TI - Taurocholate depolarizes rat hepatocytes in primary culture by increasing cell membrane Na+ conductance. AB - Rat hepatocytes in primary culture were impaled with conventional microelectrodes. Addition of 5-100 mumol/l taurocholate led to a slowly developing depolarization that was maximal at 50 mumol/l (10.5 +/- 1.5 mV, n = 15) and not reversible. The effect was Na+ dependent and decreased in cells preincubated with 1 mumol/l taurocholate. Increasing external K+ tenfold depolarized the cells by 12.3 +/- 2.3 mV under control conditions and by 6.3 +/- 1.2 mV with 50 mumol/l taurocholate present (n = 7). Depolarization by 1 mmol/l Ba2+ was 7.6 +/- 0.8 mV and 6.0 +/- 0.7 mV (n = 9) before and after addition of taurocholate, respectively. Cable analysis and Na+ substitution experiments reveal that this apparent decrease in K+ conductance reflects an actual increase in Na+ conductance: in the presence of taurocholate, specific cell membrane resistance decreased from 2.8 to 2.3 k omega x cm2 x Na+ substitution by 95% depolarized cell membranes by 8.9 +/- 2.9 mV (n = 9), probably due to indirect effects on K+ conductance via changes in cell pH. With taurocholate present, the same manoeuvre changed membrane voltages by -0.8 +/- 2.6 mV. When Na+ concentration was restored to 100% from solutions containing 5% Na+, cells hyperpolarized by 3.5 +/- 3.6 mV (n = 7) under control conditions and depolarized by 4.4 +/- 2.9 mV in the presence of taurocholate, respectively. In Cl- substitution experiments, there was no evidence for changes in Cl- conductance by taurocholate. These results show that taurocholate-induced membrane depolarization is due to an increase in Na+ conductance probably via uptake of the bile acid. PMID- 8414904 TI - Lack of control by adrenal steroids of oxidative enzymes and Na/K-ATPase development in the rat proximal tubule. AB - The activities of citrate synthase, 3-oxoacid CoA-transferase, and Na/K-ATPase were determined in the proximal convoluted tubules (PCT) of midcortical nephrons from 16-, 21- and 30-day-old and adult rats. Enzyme microassays based on NAD amplification were run on tubule segments microdissected from lyophilized tissue sections, and the activities were expressed per unit of tissue dry weight. The activities of 3-oxoacid CoA-transferase (+ 155%) and citrate synthase (+ 44%) increased between 16 and 30 days, while no significant change in Na/K-ATPase activity occurred during this period. The results obtained in PCT from subcapsular nephrons were similar. It is concluded that active transport of Na+ coupled to mitochondrial ATP production might be mature in the PCT by the time of weaning, consistent with data on the development of Na+ reabsorption. Since adrenalectomy on day 16 induced no changes in the activities of oxidative enzymes or Na/K-ATPase on day 21 in midcortical or subcapsular PCT, the physiological rise in circulating glucocorticoids, characteristic of the weaning period, does not trigger the development of oxidative enzymes and Na/K-ATPase in the PCT of the developing rat kidney. PMID- 8414905 TI - Spatial and temporal control of intracellular free Ca2+ in chick sensory neurons. AB - Digital imaging of fura-2 fluorescence and the voltage-clamp technique were combined to study cytoplasmic free Ca2+ concentration, [Ca]i, in neurons cultured from chick dorsal root ganglia. Depolarizing pulses raised [Ca]i to a new steady state level which was achieved earlier in neurites than in the soma. The rise in [Ca]i during stimulated bursting or rhythmic activity was also faster in neurites. After stimulation [Ca]i recovered monoexponentially in the soma and biexponentially in neurites. Application of 50 mM KCl produced membrane depolarization and a concomitant increase of [Ca]i. During wash-out [Ca]i often declined to an intermediate steady-state level at which it stayed for several minutes. Thereafter the resting level of [Ca]i was quickly restored. [Ca]i recovery was delayed after treating the cell with 2 microM thapsigargin, an inhibitor of the Ca2+ pump of internal Ca2+ stores. Caffeine (10 mM) transiently increased [Ca]i. A second caffeine application produced smaller [Ca]i changes due to the prior depletion of Ca2+ stores, which could be replenished by brief exposure to KCl. Thapsigargin (2 microM) transiently increased [Ca]i both in the standard and Ca(2+)-free solution. [Ca]i transients due to caffeine and thapsigargin started in the cell interior, in contrast to [Ca]i changes evoked by membrane depolarization, which were noticed first at the cell edge. Caffeine and thapsigargin induced a transient inward current which persisted in the presence of 1 mM La3+ and in Ca(2+)-free solutions, but which was greatly diminished in Na(+)-free solutions. The effects of caffeine and thapsigargin were mutually exclusive both in the generation of [Ca]i transients and in the inward current induction. PMID- 8414906 TI - Mechanical tuning characteristics of the hearing organ measured at the sensory cells in the gerbil temporal bone preparation. AB - The micromechanical behaviour of the inner ear in response to sound stimulation was investigated in an in vitro preparation of the gerbil temporal bone. Using laser heterodyne interferometry it was possible to measure the vibration responses directly at the level of the sensory and supporting cells within the hearing organ rather than from the underlying basilar membrane as has been done in previous studies. There was a tuned mechanical response of the cellular structures within the hearing organ. The resonance frequency measured at cells in the apical (third) turn was around 200Hz. The frequency of the mechanical tuning varied along the length of the cochlea. In the second turn the resonance frequency was around 500-700Hz. The cellular response in the second turn was more sharply tuned as compared to the response in the apical turn. In both cochlear turns the amplitude of the vibratory response changed with the cellular location radially across the hearing organ. PMID- 8414907 TI - Localization of NaPi-1, a Na-Pi cotransporter, in rabbit kidney proximal tubules. I. mRNA localization by reverse transcription/polymerase chain reaction. AB - We have recently isolated from a rabbit cortex cDNA library a cDNA clone (NaPi 1), which, after in vitro transcription (cRNA) and injection into Xenopus laevis oocytes, expresses Na-dependent Pi uptake [Werner A, et al. (1991) Proc Natl Acad Sci USA 88:9608-9612]. The aim of the present work was to study the nephron location of the NaPi-1-related mRNA(s) by combining nephron microdissection procedures, reverse transcription (RT) and amplification of the resultant cDNA by the polymerase chain reaction (PCR). RT-PCR using NaPi-1-specific primers (different combinations) and either total kidney cortex RNA or microdissected proximal tubule segments resulted in two PCR products, both of approximately the expected length (but differing by about 30 base pairs). Restriction-enzyme analysis and nucleotide sequencing confirmed that both PCR products are related to NaPi-1 and that the "longer" PCR product has an insert of 26 base pairs containing an AluI restriction site. Nephron microdissection documents expression of NaPi-1-related mRNA(s) in superficial and deep proximal tubules (S1, S2 and S3 segments) and their absence in glomeruli, thin descending limb and thick ascending limbs of Henle's loop, distal convoluted tubules and cortical and inner medullary collecting ducts. These experiments suggest a "microheterogeneity" of NaPi-1-related mRNA(s) (which is not detected in Northern blot analysis) and proximal tubular expression of NaPi-1. PMID- 8414908 TI - Localization of NaPi-1, a Na/Pi cotransporter, in rabbit kidney proximal tubules. II. Localization by immunohistochemistry. AB - Polyclonal antibodies have been raised against a C-terminal peptide of NaPi-1, a recently cloned Na-Pi cotransport system of rabbit kidney cortex with a predicted (unglycosylated) molecular mass of 52 kDa. By Western blot analysis using brush border membranes isolated from rabbit kidney cortex, two proteins with apparent molecular masses of 64 kDa and 35 kDa were specifically recognized (peptide protectable) by the antiserum obtained. The 64-kDa protein was found to migrate in parallel with the luminal membrane during separation by free-flow electrophoresis of brush-border and basolateral membranes. In immunofluorescence studies using cryostat sections of rabbit kidney, specific binding of antibodies was observed in proximal tubules (including S1, S2 and S3 segments) of superficial and deep nephrons. Anti-(NaPi-1)-antibody-mediated fluorescence was restricted to the brush border of proximal tubular cells. No specific immunoreaction was observed in other tubular segments. The results suggest that the native NaPi-1-related protein (Na-Pi cotransport system) has an apparent molecular mass of 64 kDa and is uniformly expressed in the apical membrane of proximal tubules of all nephron generations in the rabbit kidney. Immunohistochemical localization of the Na-Pi cotransport system NaPi-1 confirms the segmental localization within the nephron of NaPi-1-related mRNA as revealed by the reverse transcriptase/polymerase chain reaction (see preceding paper). PMID- 8414909 TI - Effect of short-chain fatty acids on cell volume and intracellular pH in rat distal colon. AB - Superfusion of isolated crypts from the rat colon with sodium-butyrate-containing solutions induced an increase in the crypt diameter indicating a swelling of the crypt cells. The response to butyrate (50 mmol l-1) was not uniform along the crypt axis, the most pronounced swelling being observed in the upper third of the crypt. The butyrate effect was concentration-dependent and was completely suppressed by amiloride, suggesting that it is caused by activation of the Na+/H+ exchanger. Acetate, propionate and isobutyrate had a similar action. In HEPES buffered solution the butyrate-induced change in cell volume was monophasic, i. e. only a swelling took place, whereas in HCO3- buffer it was biphasic, i. e. swelling was followed by a regulatory volume decrease. This decrease was suppressed by K+ and Cl- channel blockers as well as inhibitors of leukotriene synthesis. Measurements of intracellular pH with the fluorescent dye 2',7'-bis(2 carboxyethyl)-5(6)-carboxyfluorescein (BCECF) revealed that butyrate induced an acidification of the cell, which was stronger in HEPES than in HCO3- buffer. Estimation of Na+/H+ exchange activity, tested as recovery of intracellular pH from an acid load via an NH4Cl prepulse, revealed a much lower Na+/H+ exchange activity in the fundus region compared to the upper third of the crypt. The smaller volume response evoked by butyrate in the fundus region probably reflects the smaller Na+/H+ activity compared to the more differentiated cells near the opening of the crypt. It is concluded that cell swelling caused by short-chain fatty acids is a physiological stimulus for volume regulation. This response is restricted to the more differentiated cells. PMID- 8414910 TI - Renal and single-nephron function is comparable in thiobutabarbitone- and thiopentone-anaesthetised rats. AB - The thiobutabarbitone(TB, Inactin)-anaesthetised rat is an extremely widely used preparation for the study of renal function at the whole-organ and nephron levels. The recent withdrawal of TB from the market has made it essential to find an anaesthetic producing experimental conditions as similar as possible to TB to allow comparison of past and future data. Blood gas analysis, clearance and micropuncture studies were therefore performed in rats anaesthetised with TB or the related thiobarbiturate thiopentone (TP) (both 100 mg/kg body weight) to establish whether the latter meets this requirement. Both barbiturates caused similar transient respiratory depression and acidosis. Mean values (TP versus TB) over the total 8-h observation period for glomerular filtration rate (0.94 versus 1.05 ml/min), urine flow (3.8 versus 4.4 microliters/min) and K+ excretion (0.98 versus 1.18 mumol/min) were slightly lower (P < 0.05) in TP rats, whereas renal blood flow (6.26 versus 6.24 ml/min), filtration fraction (0.31 versus 0.34) and Na+ excretion (0.11 versus 0.098 mumol/min) did not differ. The single-nephron filtration rate (SNGFR) (42.1 versus 41.1 nl/min) and fractional reabsorption (42% versus 47%), both measured in the proximal tubule, did not differ, although in the TP group SNGFR rose with time (4.4%/h) whereas the fractional reabsorption did not change significantly; in the TB group SNGFR was constant but fractional reabsorption declined with time (1.5%/h). Fractional reabsorption up to the distal convoluted tubule declined with time, this was more pronounced in the TP group. SNGFR measured at this site did not differ between TP and TB (30.3 versus 30.1 nl/min) but increased with time with TP (2.7%/h).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414911 TI - Intracellular calcium ions activate a low-conductance chloride channel in smooth muscle cells isolated from human mesenteric artery. AB - Calcium-activated chloride currents were studied by the patch-clamp technique in vascular smooth muscle cells (VSMC) isolated from human mesenteric arteries. Bath application of 20 mM caffeine caused the cell membrane to depolarize by a calcium activated inward current that peaked to -654 +/- 230 pA (holding potential -50 mV). Cell-attached, at the same time inwardly directed single-channel currents were detected with an amplitude of -0.22 pA. In open-cell-attached patches channel activity was triggered by elevating [Ca2+]i to 10 microM. At -60 mV the mean amplitude of the current was -0.24 pA and the mean open time of the channels was 28 ms. Plotting the amplitude of the current versus the test potential yielded a single-channel conductance of 2.8 +/- 0.5 pS. The currents disappeared when [Cl-] was reduced from 150 mM to 5 mM at the cytosolic side of the inside out patch at a holding potential of -60 mV (calculated reversal potential -58 mV) suggesting that the calcium-activated current was a chloride current. This suggests that, in human mesenteric VSMC, elevation of [Ca2+]i activates a low conductance chloride channel, which may mediate the agonist-induced depolarization of the cell membrane. PMID- 8414912 TI - Spike conduction properties of T-shaped C neurons in the rabbit nodose ganglion. AB - The electrical activity of C-type neurons was recorded intracellularly in the rabbit nodose ganglion maintained in vitro. The initial segment of their axon is spirally wound close to the cell body and a primary branching point divides it into a central process (CP) projecting to the nucleus of solitary tract in the medulla oblongata and a peripheral process (PP) which conveys sensory inputs from the viscera. Stimulation of the CP induced either somatic ("S") spikes or low amplitude axonal ("A") spikes ("A1" or "A2"). In some cases abrupt changes in the latency of "S" or "A" spikes (jumps) were observed by gradually increasing the stimulus intensity. They are discussed in relation to a secondary branching on the central axon located inside or near the ganglion. Collision experiments showed that antidromic "A" spikes are blocked at the primary bifurcation of the axon (T-shaped neuron). Stimulation of the PP induced either "S" spikes or high amplitude "A" spikes ("A3" or "A4"). Orthodromic spikes could be blocked either before or after the primary bifurcation. When blocking occurs after the bifurcation on the stem axon, the spike can invade the central axon without invading the soma. The study of the refractory periods of the two processes and the application of high frequency stimulation showed that the PP allows higher frequencies than the soma and the CP, and thus that branching and the CP act as low-pass filters. These data support the view that the primary branching point and the CP of these T-shaped cells represent a strategic area to modulate visceral afferent messages. PMID- 8414913 TI - Molybdenum uptake through the anion exchanger in human erythrocytes. AB - Human red blood cells were incubated in the presence of Na2MoO4 and the initial rate of molybdenum uptake was measured by using inductively coupled plasma emission spectroscopy. About 99% of molybdenum uptake was inhibited by DIDS or by SITS. DIDS-sensitive molybdenum uptake was inhibited by external chloride, bicarbonate, sulphate and phosphate in the range of concentrations previously described for anion carrier fluxes. Trace elements, previously described to be translocated by the anion carrier, i.e. copper, zinc and cadmium, slightly inhibited molybdenum uptake. Molybdenum uptake was strongly stimulated by acidification, suggesting that the monovalent HMoO4- anion species, which is formed in acidic media (pK approximately 4.1), can be more rapidly translocated than the divalent anion complex MoO4(2-), which is the predominant form at physiological pH. In conclusion, the anion carrier can catalyse rapid molybdenum movements across red cells membranes. This supports previous reports of an enterohepatic circulation of molybdenum, with red blood cells acting as molybdenum carrier between the intestine and the liver. PMID- 8414914 TI - Transcellular sodium transport and basolateral rubidium uptake in the isolated perfused cortical collecting duct. AB - The relation between transcellular Na+ absorption, intracellular Na+ concentration and Na+/K(+)-ATPase activity (the last estimated by the rubidium uptake across the basolateral cell membrane) was examined in the different cell types of the rabbit cortical collecting duct (CCD). Experiments were performed on isolated perfused CCD in which Na+ absorption was varied by perfusing the tubule with solutions containing different Na+ concentrations (nominally Na(+)-free, 30 mM and 144 mM). Experiments were terminated by shock-freezing the tubules during perfusion. Precisely 30 s before shock-freezing, the K+ in the bathing solution was exchanged for Rb+. Intracellular element concentrations, including Rb+, were determined in freeze-dried cryosections of the tubules using energy-dispersive X ray analysis. Increasing Na+ concentration in the perfusion solution caused significant rises in intracellular Na+ concentration and Rb+ uptake of principal cells. Principal cell Na+ and Rb+ concentrations were 7.8 +/- 0.9 and 7.0 +/- 0.8 mmol/kg wet weight respectively, when the perfusion solution was Na(+)-free, 10.1 +/- 0.7 and 11.6 +/- 0.6 mmol/kg wet weight with 30 mM Na+ in the perfusion solution, and 14.5 +/- 1.5 and 14.9 +/- 0.9 mmol/kg wet weight with 144 mM Na+ in the perfusion solution. In contrast, a comparable relationship between lumen Na+ concentration, intracellular Na+ concentration and basolateral Rb+ uptake was not seen in intercalated cells. These results support the notion that principal, but not intercalated, cells are involved in transepithelial Na+ absorption. In addition, the data demonstrate that apical Na+ entry and basolateral Na+/K(+) ATPase activity are closely coupled in principal cells of the rabbit CCD.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414915 TI - Tidal variation of pulmonary blood flow and blood volume in piglets during mechanical ventilation during hyper-, normo- and hypovolaemia. AB - Effects of changes in blood volume on changes in pulmonary blood flow and pulmonary blood volume during the ventilatory cycle during mechanical ventilation with a positive end-expiratory pressure of 2 cm H2O were determined in six pentobarbital anaesthetized, curarized pigs weighing about 10 kg. Haemodynamic variables were analysed for each cardiac cycle in eight ventilatory cycles in four consecutive series under hyper-, normo- and hypovolaemic conditions. Cardiac output was highest in hypervolaemia. Compared with normo- and hypovolaemia, it decreased less during inflation, due to a smaller rise in central venous pressure and presumably a larger filling state of the venous system. The smaller decrease in right ventricular output in hypervolaemia coincided with a larger fall in transmural central venous pressure (right ventricular filling pressure), due to right ventricular action at a higher, less steep part of its function curve. The difference between right ventricular-output (electromagnetic flow measurement) and left ventricular-output (pulse contour) indicated changes in pulmonary blood volume. In hypervolaemia less blood shifted from the pulmonary circulation into the systemic system during inflation than in normo- and hypovolaemia. This difference can be explained by two mechanisms namely, the smaller fall in input into the pulmonary vascular beds and a smaller pulmonary vascular volume decrease as a result of transmural pressure fall at a steeper part of the pressure-volume curve. PMID- 8414916 TI - Arterial baroreflex inhibition by midbrain periaqueductal grey in anaesthetized rats. AB - Midbrain periaqueductal grey (PAG) provokes the defense reaction when stimulated. The present study was conducted to determine whether, and how, the PAG produces baroreflex inhibition, a feature characterizing the hypothalamic defense reaction. In chloralose-urethane anaesthetized rats, baroreflex vagal bradycardia and baroreflex hypotension were provoked by aortic depressor nerve stimulation. When the PAG was electrically stimulated baroreflex vagal bradycardia was remarkably suppressed; suppression of baroreflex hypotension was observed following bilateral vagotomy. In contrast, chemical stimulation of the PAG by D,L homocysteic acid microinjection markedly suppressed baroreflex vagal bradycardia but only minimally suppressed baroreflex hypotension. These findings suggest that whereas overall PAG stimulation inhibits not only cardiac but also vascular components of baroreflexes, inhibition of the latter component either depends largely on activation of passing fibers or requires recruitment of a larger number of PAG cell bodies. PAG inhibition of baroreflex vagal bradycardia was not affected following spinal cord transection at C1, indicating that the inhibition was exclusively central in origin and not due to peripheral, prejunctional inhibition of vagal acetylcholine release by increased cardiac sympathetic nerve activities. The PAG inhibition of baroreflexes was greatly attenuated following electrolytic as well as chemical destruction of the parabrachial region. On the other hand, when the PAG was extensively lesioned, baroreflex inhibition produced by hypothalamic defense area stimulation was markedly diminished. PAG excitation thus causes powerful inhibition of arterial baroreflexes which is mediated by the parabrachial region; the PAG also mediates a major fraction of hypothalamic inhibition of the baroreflexes. PMID- 8414917 TI - Acute adaptation and resetting of the baroreflex control of vascular resistance in the canine hindquarters and mesentery. AB - To determine whether acute adaptation and resetting occur in the baroreflex control of regional vascular resistance, experiments were conducted in anesthetized and vagotomized dogs. The carotid sinuses were vascularly isolated to regulate the carotid sinus pressure (CSP) in an open-loop fashion. The hindquarters (n = 12) and mesenteric (n = 10) beds were perfused with constant flow and arterial perfusion pressures (HPP and MPP) were used to reflect changes in hindquarters and mesenteric resistance respectively. We first observed alterations in HPP and MPP during the course of CSP holding (conditioning pressure) at various levels for 15 min. Thereafter, the CSP was lowered to 50 mm Hg and increased stepwise to obtain the CSP-HPP and CSP-MPP baroreflex function curves. In experiments in the hindquarters bed, HPP stabilized at an average of 104.7 mm Hg during the initial conditioning pressure at 100 mm Hg. When conditioning pressure decreased to 50 mm Hg, the HPP increased to 125.5 mm Hg and then gradually declined to a steady level (115.6 mm Hg) in 5 min. An increase in conditioning pressure from 100 to 150 mm Hg caused HPP to decrease to 54.8 mm Hg followed by an upward adaptation to a steady level (80.2 mm Hg) in 5 min. The CSP/HPP curves constructed from the CSP step protocol were also affected by conditioning pressure. There were significant increases in the threshold and saturation pressures as conditioning pressure was elevated. However, the resetting was characterized by a parallel shift of the CSP/HPP curves without significant changes in baroreflex gain or sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414918 TI - Modulation of glycogen metabolism of rat skeletal muscles by endurance training and testosterone treatment. AB - The effects of training and/or testosterone treatment on glycogen content and the activities of glycogen synthase, glycogen phosphorylase, and fructose-6-phosphate kinase were studied in extensor digitorum longus (EDL) and soleus muscles of intact adult female rats. One group of rats remained sedentary, whereas another group was trained for 7 weeks. Thereafter, both the sedentary and trained rats were subdivided into two control and four testosterone-treated subgroups. Testosterone was administered by a silastic implant. Training was continued for 2 weeks. On the final day of the experiment rats from one trained control and one trained testosterone-treated subgroup ran for 60 min submaximally. Upon testosterone treatment of sedentary rats the glycogen concentration was not changed. However, in the soleus, but not in the EDL, the glycogen content was increased by training (P < 0.05) which could, at least partly, be explained by a decrease in activity of active glycogen phosphorylase (P < 0.05). In the EDL of trained rats testosterone treatment increased glycogen content significantly by both an increase in activity of active glycogen synthase and a decrease in activity of active glycogen phosphorylase (P < 0.05). In the EDL and soleus of testosterone-treated animals from the exercised subgroup a significant sparing of glycogen was observed, which could be explained by an increase in activity of active glycogen synthase and, in the soleus, could also be explained by a concerted decrease in active glycogen phosphorylase (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414919 TI - Electrogenic Na+/K(+)-transport in human endothelial cells. AB - Na+/K+ pump currents were measured in endothelial cells from human umbilical cord vein using the whole-cell or nystatin-perforated-patch-clamp technique combined with intracellular calcium concentration ([Ca2+]i) measurements with Fura-2/AM. Loading endothelial cells through the patch pipette with 40 mmol/l [Na+] did not induce significant changes of [Ca2+]i. Superfusing the cells with K(+)-free solutions also did not significantly affect [Ca2+]i. Reapplication of K+ after superfusion of the cells with K(+)-free solution induced an outward current at a holding potential of 0 mV. This current was nearly completely blocked by 100 mumol/l dihydroouabain (DHO) and was therefore identified as a Na+/K+ pump current. During block and reactivation of the Na+/K+ pump no changes in [Ca2+]i could be observed. Pump currents were blocked concentration dependently by DHO. The concentration for half-maximal inhibition was 21 mumol/l. This value is larger than that reported for other tissues and the block was practically irreversible. Insulin (10-1000 U/l) did not affect the pump currents. An increase of the intracellular Na+ concentration ([Na+]i) enhanced the amplitude of the pump current. Half-maximal activation of the pump current by [Na+]i occurred at about 60 mmol/l. The concentration for half-maximal activation by extracellular K+ was 2.4 +/- 1.2 mmol/l, and 0.4 +/- 0.1 and 8.7 +/- 0.7 mmol/l for Tl+ and NH4+ respectively. The voltage dependence of the DHO-sensitive current was obtained by applying linear voltage ramps. Its reversal potential was more negative than -150 mV. Pump currents measured with the conventional whole-cell technique were about four times smaller than pump currents recorded with the nystatin-perforated-patch method.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414920 TI - Effect of vasoactive intestinal peptide, carbachol and other agonists on the membrane voltage of pancreatic duct cells. AB - The regulation of pancreatic exocrine secretion involves hormonal, neural and neurohormonal components. Many agonists are known to be effective in pancreatic acinar cells, but less is known about the ducts. Therefore, we wanted to investigate the influence of various agonists on isolated perfused pancreatic ducts and, as a physiological response, we measured the basolateral membrane voltage of the duct cells (Vbl) with microelectrodes. Pancreatic ducts were dissected from pancreas of normal rats and bathed in a HCO(3-)(-containing solution. Under control conditions, the average Vbl was between -50 and -70 mV. Vasoactive intestinal peptide (VIP) and carbachol (CCH) reversibly depolarized Vbl when applied to the bath. VIP (9 x 10(-9) mol/l) depolarized Vbl from -72 +/- 3 mV to -53 +/- 3 mV (n = 20) and CCH (10(-5) mol/l) from -62 +/- 3 to -35 +/- 4 mV (n = 10). Furthermore, a decrease of the Cl- concentration in the lumen led to an increase of VIP-induced depolarization of Vbl, suggesting that a luminal Cl- conductance was increased. Cholecystokinin (CCK, 10(-10)-10(-7) mol/l) and bombesin (10(-8), 10(-5) mol/l), which stimulate pancreatic exocrine secretion in acini or whole glands, showed no significant effect on Vbl of the duct cells tested in our preparation (n = 7, 6). Neurotensin (10(-8) mol/l) had a marked depolarizing effect in two out of ten cases; Vbl depolarized from about -65 mV to -29 mV and the effect was reversible. Substance P (2 x 10(-7) mol/l), alone or in combination with secretin, had no effect on Vbl of the tested duct cells (n = 11).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414921 TI - Increase in cytosolic Ca2+ regulates exocytosis and Cl- conductance in HT29 cells. AB - Increases of cytosolic Ca2+, as occur with agonists such as ATP, neurotensin (NT), hypotonic cell swelling and ionomycin, enhance the membrane conductance (GM) and hence the input conductance (GI) of HT29 cells. In the present study we have examined whether these increases in GM are paralleled by exocytosis. To this end the membrane capacitance (CM) of HT29 cells was measured by patch clamp techniques. Two methods to monitor CM were used: a direct method (DM) and a phase tracking method (PTM). With the DM the following results were obtained. NT (10( 8) mol/l, n = 9) increased GM and CM significantly from 2.4 +/- 0.3 nS and 23.5 +/- 3 pF to 32 +/- 8 nS and 27.3 +/- 3.1 pF respectively. ATP (10(-4) mol/l, n = 29) had a very similar effect. GM and CM were increased from 5.7 +/- 1 nS and 36 +/- 4.4 pF to 111 +/- 21 nS and 44 +/- 5.4 pF respectively. Hypotonic cell swelling (160 mosmol/l, n = 18) had a comparable effect: GM and CM were increased from 4.9 +/- 1 nS and 30 +/- 4.1 pF to 46 +/- 10 nS and 37 +/- 4.9 pF respectively. Ionomycin (10(-7) mol/l, n = 4) gave similar results. With the PTM it was possible to monitor the rapid changes in GM and CM, as they were induced by ATP (n = 42) and NT (n = 29), with high time resolution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414922 TI - Beta adrenergic and muscarinic receptors in compensatory cardiac hypertrophy of the adult rat. AB - The beta adrenergic (beta AR) and muscarinic (MR) receptors have been quantitated in parallel, using 125I-pindolol and 3H-quinuclidinylbenzilate, in a model of compensatory left ventricular (LV) hypertrophy (LVH), which developed in rats 4-6 weeks after an abdominal aortic stenosis. Since aortic banding resulted in a pronounced LVH of 62%, the results were expressed both in terms of density (fmol/mg protein) and quantity (fmol per LV). In addition, competition curves using either a specific beta 1-antagonist or isoproterenol or carbachol allowed the determination of the two beta AR subtypes and of the low and high affinity sites (defined by the inhibitory constant Ki) for both beta 1AR and MR. In LVH, receptor density decreased for each of total beta AR, beta 1AR subtype, high affinity (Ki 6-8 nM) beta 1AR sites (from 26 +/- 2 to 19 +/- 3 fmol/mg protein, P < 0.05), total MR and high affinity (Ki 12 nM) MR sites (from 63 +/- 6 to 40 +/- 4 fmol/mg protein, P < 0.001). The beta AR and MR densities dropped in parallel so that the MR/beta AR ratio remained unchanged. In sharp contrast (because the LVs were bigger) the quantities of total beta AR, beta 1AR subtype, beta 1AR high affinity sites, total MR and MR high affinity sites per LV were unmodified.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8414923 TI - A linear 16 pS Cl- channel in the apical membrane of Caco-2 cells. AB - A linear Cl- channel with a mean conductance of 16.3 +/- 0.4 pS (n = 7), as determined for inwardly directed currents in the cell-attached mode, was identified by the patch clamp technique in the apical membrane of Caco-2 cells in the early stage of postconfluential differentiation. This channel exhibited a low spontaneous activity (in 3 out of 137 experiments) and was not activated by strong polarization of the holding potential. After stimulation of the cAMP signal transduction cascade at different levels the 16 pS Cl- channel was activated in 7 out of 67 experiments during continuous cell-attached recording and remained active after excision of the patch. This channel differed markedly from both a linear low conductance Cl- channel and an intermediate conductance, outwardly rectifying (ICOR) Cl- channel, already reported for Caco-2 and other colonic cell lines. Thus the 16 pS channel may represent a further type of apical Cl- channel in colonocytes. PMID- 8414924 TI - Brain-stem cooling by face fanning in severely hyperthermic humans. PMID- 8414925 TI - [Morphological analysis of the secondary pulmonary lobule with inflated fixed lung specimen. Part 2: Three-dimensional measurement of intralobular structures]. AB - In this study, we made three-dimensional measurements of intralobular structures in normal and diseased conditions using inflated fixed lung specimens. Lung blocks obtained from the subpleural and hilar areas were cut into slices 0.5 mm thick and radiographed. After tracing intralobular structures on these radiograms, trace data were transferred to a three-dimensional computer system and three-dimensional measurement was made. Autopsied lungs with diseased conditions including idiopathic pulmonary fibrosis, aging lung and panacinar emphysema were also used. The distance from the tip of the intralobular arteriole accompanying the terminal bronchiole to the corresponding lobular border was 2.4 +/- 0.5 mm in the subpleural area and 2.1 +/- 0.5 mm in the hilar area. The corresponding values in emphysema, aging lung and idiopathic pulmonary fibrosis were 4.1 +/- 0.6, 3.4 +/- 0.4 and 1.4 +/- 0.2 mm, respectively. In most of the lobules, the volume ranged from 300 to 600 mm3 and was closely related in linear fashion with the number of terminal bronchioles. Branching angles of the pulmonary artery in the lobule ranged from 80 to 90 degrees, and there was no relationship to the pattern of branching. This information about the arrangement of pulmonary lobular structures in normal and diseased conditions will help in the interpretation of HRCT images of the lung. PMID- 8414926 TI - [Rounded atelectasis with emphasis on its wide spectrum]. AB - While rounded atelectasis (RA) is considered to be rather common in the United States and Europe, the total number of RA cases reported from Japan still remains approximately 30. We have long been aware that there are many variations in the radiographic appearance of so-called RA and that RA has never been clearly defined. We retrospectively reviewed 22 cases collected as RA and its variants from several institutions. We defined RA as "peripheral atelectasis mimicking tumor secondary to shrinkage or bending of the pleura of various degrees, and accompanied by lung distortion." The diagnostic criteria of typical RA include (1) peripheral tumoral shadow in contact with pleural effusion or thickened pleura, (2) acute angle between the pleura and the shadow, (3) convergence of the pulmonary vessels and bronchi and (4) volume loss of the affected lobe. However, there are cases which lack some of these criteria but are considered to be included in the broad category of RA. We propose that RA should be considered to be an entity having a wide spectrum. Typical lesions showing "cranial tilting" of Hanke are on one side of the spectrum and small linear or strand shadows extending from the thickened pleura are on the other. PMID- 8414927 TI - [Visualization of coronary arteries with helical scanning CT: diastolic reconstruction]. AB - The purpose of this study was the visualization of coronary arteries by the diastolic reconstruction method with helical scanning (HES) CT. We chose diastolic phase images from the image data acquired with the HES technique using a continuous nutate-rotate fast CT scanner, because few motion artifacts are induced by cardiac pulsation during the slow filling diastolic phase. We assessed coronary diseases using this method based on HES-CT, and evaluated its clinical effectiveness. We also studied the clinical value of multiplanar reconstruction and three-dimensional surface reconstruction. The subjects consisted of 31 patients with coronary disease, aged 41-77 years. When a diastolic reconstruction HES-CT was employed, the average rate of visualization of the coronary arteries was 72%. An examination can be completed during a 30-second single breath-hold, permitting the acquisition of coronary artery images with excellent continuity. Calcified coronary arteries can be identified. PMID- 8414928 TI - [X-ray VTR analysis of esophagocardiac motion in cases of cervical dysphagia]. AB - Using X-ray VTR motion analysis to diagnose a total of 103 cervical dysphagial cases, we obtained the following conclusions. Among these dysphagial cases, there were only 32 cases (31%) of clear organic or functional disease which we commonly considered to be the cause of dysphagia, and there were no less than 71 cases (69%) in which we could not identify the cause of the dysphagia. In 29 of these 71 cases, recording conditions of whole esophageal systole and diastole through the passage of contrast medium (Barium) were insufficient. We compared the remaining 42 cases with 42 normal cases, focussing on the lower esophagocardiac movement. In these dysphagial cases, angle of barium inflow tended to steepen statistically, and Barium discharge of the ampulla was tended towards incomplete discharge. In conclusion, this method is thought to contribute to the diagnosis of dysphagial cases except clear organic or functional diseases which we commonly consider to be the cause of the dysphagia. PMID- 8414929 TI - [Evaluation of the blood flow in lung disease with color Doppler flow imaging]. AB - Blood flow within the lesions was evaluated in 6 patients of lung cancer and in 4 patients with pulmonary tuberculosis with color Doppler flow imaging. Three different signal types were noted: pulsatile signal was detected in all patients with lung cancer and in 2 patients with pulmonary tuberculosis, continuous signal was only detected in 3 patients with lung cancer, triphasic signal was detected in 4 patients with pulmonary tuberculosis and in 2 patients with lung cancer. It was seemed to be that triphasic signal was blood flow signal in peripheral vessel of pulmonary vein and color Doppler flow imaging was useful for evaluation of blood flow patterns in lung disease. PMID- 8414930 TI - [Treatment results of breast cancer patients who hoped for breast preservation]. AB - From 1983 to August 1990, 143 patients (148 breasts) were treated with breast conservation therapy in our department. Age ranged from 27 to 75 years (median: 45 years). Forty-one were stage I, 91 were stage II, 6 were stage III, and 5 were stage IV. Surgery consisted of lumpectomy and axillary dissection up to level II. When axillary lymph node(s) were pathologically involved (n(+)), chemotherapy (CMF) was performed. Radiation dose was 50 Gy to the entire breast, and booster irradiation was not performed. After October 1989, patients without palpable axillary node(s) underwent lumpectomy without axillary dissection. Instead, almost all patients started to receive chemotherapy, and not only the breast but also lymph node areas were also included in the radiation field. In 143 patients received conservation therapy, 5-year survival rate was 94%, and 5-year freedom from relapse rate was 81%, respectively. Microscopic surgical margin was positive in 39%. Excluding 5 stage IV patients, 14 relapsed out of 138 patients. Initial relapse sites were as follows: the breast in 3, the axilla 2, the distant organ in 10 patients. One patient relapsed in the axilla and the distant site simultaneously. Distant sites were the lung (4), the bone (4), the bone and the other organs (2). Three stage II patients showed in-breast recurrences. Microscopic surgical margins at initial lumpectomy were positive in one and negative in other 2 patients. One patient was salvaged with re-lumpectomy, and no relapse was seen after that. PMID- 8414931 TI - [Clinical evaluation of myocardial perfusion imaging with 99mTc-tetrofosmin]. AB - Myocardial perfusion imaging with Tc-99 m-1,2-bis (bis (2-ethoxyethyl) phosphino) ethane (Tc-99 m-tetrofosmin) was performed in 26 patients with ischemic heart disease (IHD) or suspected to have IHD. The agent was administered both at rest and at peak exercise. Exercise and rest studies were performed on the same day or on different days. The injected dose ranged from 296 MBq-740 MBq. Image quality was adequate for diagnosis, and was superior to that of T1-201 scintigraphy. Results from the tetrofosmin imaging corresponded well with findings of T1 scintigraphy, with a segmental concordance of 87%. Sensitivity and specificity were 70% and 93%, respectively. Our results suggested that tetrofosmin could be a promising Tc-99m labeling agent for the evaluation of myocardial perfusion. PMID- 8414932 TI - [Evaluation of the blood flow in tumors before and after irradiation with color Doppler flow imaging]. AB - The blood flow in tumors of three patients with lung cancer, cancer in oral cavity and metastatic lymphnode was respectively evaluated before and after irradiation with color Doppler flow imaging. The arterial pulsating flow in lung cancer disappeared after 30 Gy irradiation and that in cancer of oral cavity disappeared after 15 Gy irradiation with intraarterial infusion chemotherapy. But the arterial pulsating flow in metastatic lymphnode was detected during irradiation and disappeared after 40 days post- irradiation. Therefore color Doppler flow imaging seemed to be useful for evaluation of the blood flow in tumors before and after irradiation. PMID- 8414933 TI - [Superselective bronchial arterial infusion therapy with cisplatin and epirubicin hydrochloride, mitomycin C-iohexol-Lipiodol emulsion (EMILE) for hilar lung adenocarcinoma: preliminary clinical experience]. AB - A case of hilar lung adenocarcinoma was treated by superselective bronchial arterial infusion therapy with cisplatin and epirubicin hydrochloride, mitomycin C-iohexol-Lipiodol emulsion (EMILE) using Tracker -18 infusion catheter. The tumor size was reduced on follow-up CT scans. However, EMILE was also distributed to nontumorous lung tissues around the tumor, and a shrinkage of the right upper lobe and elevations of the right hilus and diaphragm followed. No major complaints and clinical complications during and after the treatment occurred. This therapy was safe and effective for local tumor reduction in a case of hilar lung adenocarcinoma. PMID- 8414934 TI - [Dynamic MR imaging of the pituitary gland by the fast spin echo (RARE) sequence]. AB - The Fast Spin Echo (RARE: Rapid Acquisition with Relaxation enhancement) sequence for the dynamic MRI of the pituitary gland was performed in 18 patients suspected of the intracranial lesions. The SNR of the plain image of 5 pituitary glands was measured on the FSE 400 and 200/17/8/2 (TR/effective TE/echo train length/excitation) and the spin echo 100/11/2 (TR/TE/excitation) sequence. The FSE (TR = 400) provided the highest SNR than others, The FSE sequence was able to acquire increased spatial resolution and reduced acquisition time, and was the significant sequence for the dynamic MRI of the pituitary gland. PMID- 8414935 TI - [Clinical trial of IVC filter to temporary placement]. AB - We developed a new inferior vena caval filter to prevent pulmonary embolism for a patient with IVC thrombus. This filter made by covering top half of Dotter Intravascular Retriever catheter with Dacron mesh sheet, was placed in IVC through a 10 F sheath from right jugular vein. In one case, after anticoagulant therapy and thrombectomy using balloon catheter, many free-floating thrombus were trapped in this filter and were taken out from the IVC. This filter was useful for preventing pulmonary embolism when removing thrombus in IVC, iliac and lower extremity veins. We propose to call this device "Hino's filter". PMID- 8414936 TI - [Apoptosis in radiosensitive malignant tumors]. AB - This study reported the radiation induced apoptosis in 6 cases with malignancies that were observed rapid tumor regression with radiotherapy up to 10 Gy. Microscopic examinations of 6 cases revealed the tumor cell number reduction and the appearance of apoptotic cells in the irradiated cases. It is suggested that the radiation-induced apoptosis plays an important role in rapid tumor regression by radiotherapy and occurs not only in lymphoma which has been recently reported in vitro but also in carcinoma. PMID- 8414937 TI - [Morphological analysis of the secondary pulmonary lobule with inflated fixed lung specimen. Part 1: Evaluation of the branching pattern and course of the intralobular pulmonary artery]. AB - This study was undertaken to clarify the relationship between the pulmonary artery and airway by three-dimensional analysis of inflated fixed lung specimens. Lung blocks obtained from the subpleural and hilar areas were cut into slices 0.5 mm thick and radiographed. After tracing intralobular structures on these radiograms, trace data were transferred to a three-dimensional computer system. By observing each specimen and the three-dimensional reconstructed images, we analyzed the intralobular structures and the course of the pulmonary artery in particular. The lobular and intralobular pulmonary arteries closely followed the corresponding bronchial tree in the subpleural area. In contrast, lung areas supplied by the daughter bronchus showed a very different appearance. In these areas, the pulmonary artery and bronchus were usually separated from each other until the tip of terminal bronchiole. A pattern in which a single pulmonary lobule was supplied by a few pulmonary arterial branches was seen in 70% of the lobules. This information will help in the interpretation of HRCT (high resolution CT) findings related to the bronchovascular bundle especially in the area supplied by the daughter bronchus. PMID- 8414938 TI - HIV/AIDS train the trainer program for nursing home educators. PMID- 8414939 TI - It's a long story. PMID- 8414940 TI - [Who is responsible for the "wave of laparoscopy"]. PMID- 8414941 TI - [Laparoscopic surgery. A shift in paradigm?]. AB - Since it was first introduced at the beginning of the century, laparoscopy has been developed by pioneers in the field of gynaecological surgery from a diagnostic aid to a high tech tool for use in various branches of surgery. In the near future, this rapidly developing technique, with three-dimensional video and robot-assisted surgery, will require well planned theoretical and practical training. PMID- 8414942 TI - [Endocrine surgery today]. AB - Endocrine surgery is an important branch of modern surgery, and includes surgery of the thyroid and parathyroid as well as operative treatment of endocrine tumours in the adrenals, pancreas and the alimentary tract. Close cooperation between endocrine surgeons, medical endocrinologists, pathologists and radiologists is a prerequisite. The development of endocrine surgery has a long tradition in the Scandinavian countries, characterised by cooperation both in surgical training and research. PMID- 8414943 TI - [Screening for colorectal cancer--does it help?]. AB - During the last decade, evidence from retrospective as well as prospective case control studies suggest that screening for colorectal cancer in average-risk persons above 50 years of age may be worthwhile. Screening with proctoscopy may reduce the mortality from rectal cancer and screening with fecal occult blood tests (Hemoccult-II) may reduce the mortality from colorectal cancer. Final results from ongoing population studies will appear within a few years and there are reasons to believe that mortality for colorectal cancer may be reduced with at least 30 per cent by a combination of Hemoccult-II and flexible 60 cm sigmoidoscopy. PMID- 8414944 TI - [Hormone treatment in the climacteric]. AB - To improve the quality of life among women during the three decades following the menopause, every effort should be made to prevent the adverse effects of ageing. Broad joint initiatives, involving various disciplines such as, cardiology, urology, oncology, orthopaedics and gynaecology, are needed to optimise medical care during the climacteric. Oestrogen-progesterone treatment is the most important means currently available for the prevention of postmenopausal problems. This form of hormone therapy also appears to provide protection against osteoporosis and cardiovascular disease. PMID- 8414945 TI - [Peter F. Hjort--with close contact and a bird's eye view of public health. Interview by Eva Oldinger]. PMID- 8414946 TI - [Scandinavian Federation for Medical Education. Education news]. PMID- 8414947 TI - [Epidemiological research with or without registries?]. PMID- 8414948 TI - [The use of disease registries--a question of ethics]. AB - The Nordic countries have unique possibilities for performing register-based epidemiological research. Current proposals by the European Community and the Council of Europe may radically hamper such research, however. It is important that researchers enter into the debate and demonstrate the value of such epidemiological research. The article consists in an account of the registers now available that are suitable for use from the point of view of research, and some of the results obtained. The conclusion is that it would be unethical not to take advantage of the abundant source of data represented by the disease registers. PMID- 8414949 TI - [Medical birth registries a gold mine for research]. AB - Systematic monthly monitoring of the occurrence of congenital abnormalities has been practised in Norway since 1971, and has made important contributions in the field of perinatal epidemiology. The registry would also be a unique source of data for generation studies of reproductive outcome. Hitherto, little advantage has been taken of the possible uses of the registry for research into factors of importance to children's development, health and function. Important contributions can be made by using the registry in conjunction with other relevant registers within the health and social sectors. A research co-operation between the birth registers in the five Nordic countries was established in 1970. PMID- 8414950 TI - [Cancer registry and data security]. AB - Population-based cancer registration has existed in the five Nordic countries for 35-50 years. With the exception of Sweden, the Nordic cancer registries not only collect figures of annual incidence, but also serve as institutes for cancer statistics and epidemiological research. The article gives an account of the scope of research based on cancer registries in the Nordic countries, as illustrated by examples from Finland. PMID- 8414951 TI - [Ethics in a market-oriented health care system]. PMID- 8414952 TI - [Apoplexy--the acute stage]. AB - This review concerns the acute phase of stroke. It describes incidence, prevalence, etiology, diagnosis and treatment together with the possibilities for prevention. The incidence of stroke in the Danish population is about 2/1000 person years and has been largely unchanged during the last 20 years. About 85 per cent of strokes are caused by cerebral infarcts, ten percent by intracerebral haemorrhages and about five per cent by subarachnoid bleeding. The incidence increases with age. Up till age 65 years the ratio between men and women is two to one, while the ratio in the oldest age group approaches one to one. The most important risk factors for stroke are smoking, arterial hypertension, previous cerebrovascular disease, heart disease and diabetes mellitus. Till now, no treatment has been documented as effective in reducing the cerebral damage caused by acute stroke. Ongoing controlled clinical trials in the acute state of ischaemic stroke are testing the effect of thrombolytic therapy, treatment with calcium antagonists, glutamate receptor antagonists, aspirin and heparin. The general medical treatment including nursing and physiotherapy in the acute phase is described. Within recent years benefit of various strategies of stroke prevention has been documented. PMID- 8414953 TI - [Smoking habits among pregnant women in Norway--suggestions for intervention]. AB - Multiple surveys have shown that approximately 35 percent of women in Norway are daily cigarette consumers during their pregnancies. Smoking is the single most important risk factor in pregnancy leading to excessive rates of low birth weight, prenatal neonatal and infant mortality, sudden infant death syndrome and learning problems in school. Norway provides a decent prenatal care program free of charge to all pregnant women. But efforts to reduce smoking have been inadequate. A multilevel strategy including a media campaign, the policy changes recommended in the Action Plan "Smokefree Norway by the year 2000" and a standardized intervention program using generally available resources is suggested as a possible way to reduce the problem throughout the country. PMID- 8414954 TI - [Do pregnant women need supplementation of folic acid?]. AB - Neutral tube defects (NTDs) occur at an incidence of about 0.01 per cent, and in Denmark 40-50 new cases are recorded annually. Although the metabolism of folic acid increases during pregnancy, the dietary supply is marginal. Finding in recent studies suggest that folic acid supplementation as prophylactic treatment of gravidae reduces the incidence of NTDs. Folic acid prophylaxis is already being recommended as a general policy, with treatment starting before conception. PMID- 8414955 TI - [What is about Iceland--is not in Iceland]. PMID- 8414956 TI - [Human rights and medical activities--what can Scandinavian physicians do?]. PMID- 8414957 TI - [Large differences in public health among the Baltic countries]. PMID- 8414958 TI - [EEC support for education in community medicine in Hungary]. AB - Several East European countries are faced with far-reaching changes both in the health care system and in medical education. In Hungary, this is partly due to the background of political change, but also to the manifest deterioration of public health during the past 10-20 years. A Hungarian aim has been to reform both undergraduate and postgraduate medical education, with a greater emphasis on community medicine. Via its TEMPUS programme, the EC is therefore supporting the modernization of Hungarian education in community medicine. PMID- 8414959 TI - [Leukemia. History of concepts. Campaign against dogma]. PMID- 8414960 TI - Diagnosis and management of foetal thrombocytopenia. AB - Platelet counts remain stable during intrauterine life (245 +/- 65 x 10(9)/litre, mean +/- SD). Before diagnosing thrombocytopenia (< 150 x 10(9)/litre), a foetal blood sample must be checked for contamination with amniotic fluid, since even slight contamination can activate coagulation and lead to a false positive result. In this paper, we review the major causes of thrombocytopenia and discuss their pathogenesis and management. Foetal thrombocytopenia can be caused by maternal complications (immune thrombocytopenic purpura, neonatal alloimmune thrombocytopenia, gestational thrombocytopenia, preeclampsia, alloimmune haemolytic disease) or infectious diseases (toxoplasmosis, cytomegalovirus, rubella) or be of true foetal origin (chromosomal abnormalities, malformations, congenital thrombocytopenia, intrauterine growth retardation. PMID- 8414961 TI - Plasma cell leukaemia mimicking acute monocytic leukaemia in the course of multiple myeloma. AB - We report a case of acute leukaemia occurring early in the course of IgA multiple myeloma. Ultrastructural studies, immunophenotyping and karyotyping were required to identify the origin and clonality of the leukaemic cells. Although ultrastructural examination of the blast cells revealed both monocytoid and plasma cell features, all cells expressed the CD 38 antigen and intracytoplasmic kappa light chains, while karyotyping revealed a clone with numerous abnormalities, leading to the diagnosis of clonal plasma cell disease. The occurrence of leukaemia in multiple myeloma is discussed. PMID- 8414962 TI - Plasma erythropoietin in essential thrombocythaemia. AB - Plasma erythropoietin levels were frequently decreased in patients with essential thrombocythaemia (14/31 cases), as in polycythaemia vera. These two diseases or some forms of them might be two facets of a single disease process. PMID- 8414963 TI - Decreased protein S activity in sickle cell disease. AB - In sickle cell disease (SCD), vaso-occlusion is a complex process involving cellular, vascular and humoral factors and possibly thrombotic events. We studied three physiological inhibitors of the coagulation system, antithrombin III (AT III), protein C (PC) and protein S (PS), in three groups of subjects: 27 homozygous patients observed either in crisis or in a steady state, 23 heterozygous patients and 30 healthy subjects. PS study included the measurement of total and free PS antigen, PS activity and C4bBP antigen. In heterozygous subjects the results were similar to those of controls, but in homozygous subjects abnormalities of PS and to a lesser extent PC were observed. Values of PC were extremely variable with 10 cases lower than the normal range (2 SD of the mean) and 17 others within this range. In all cases total PS antigen was slightly reduced (77 +/- 18%, M +/- SD) with a more marked decrease of free antigen (59 +/ 17%) and normal values of C4bBP. Levels of PS activity were greatly reduced and lower than those of free antigen with a mean ratio of PS activity to free antigen of 0.6. These abnormalities were associated with significantly high concentrations of fibrinogen D-dimers. PS deficiency in SCD may be at least partly due to adsorption of free PS to aminophospholipids abnormally expressed on sickle cells membranes, microvesicles and activated platelets, while the discrepancy between PS activity and free antigen could reflect proteolytic inactivation of PS by traces of thrombin. PMID- 8414964 TI - Platelet production in idiopathic thrombocytopenic purpura. AB - 522 patients with chronic thrombocytopenia were studied over a period of three years. 252 of them suffered from idiopathic, acquired, thrombocytopenic purpura with excessive platelet destruction (platelet count less than 50,000 and mean survival less than 3 days). All of these cases were studied by using Indium oxinate labelled autologous platelets. The essential finding in these cases was the absent or only slight reactive hyperproduction, regardless of age and regardless of the severity of platelet destruction. No significant modification in platelet production was observed in the presence or in the absence of anti platelet antibodies. No significant difference in the haematopoietic response, which was only exceptionally greater than twice the normal value, was observed whether thrombocytopenia was or not very long-standing, very severe, or affected a child or an adult. Megakaryocytosis, assessed on smears or biopsy sections, and the mean platelet volume do not constitute parameters for measuring platelet production. These findings confirm previous findings reported in the 1970s using radioactive chromium as the tracer. They demonstrate that the regulation of megakaryocyte production differs from that of erythropoietic production. They also show that determination of platelet production based on the survival of autologous platelets is able to distinguish between excessive destruction, even in the absence of a haematopoietic response, and defective production, demonstrated when the haematopoietic response is less than 50% of the normal value. PMID- 8414965 TI - Modality treatment in adult stage I-II Hodgkin's disease with large mediastinal masses: study of 30 cases and review of the literature. AB - From January 1980 to December 1989, 30 untreated patients with supradiaphragmatic Hodgkin's disease (HD) stage IA to IIBE, presenting a mediastinal mass with mediastinal to thoracic ratio (MTR) > or = 0.33, were treated by combined modality therapy. None had staging laparotomy and the range of MTR was 0.33 to 0.60 (mean 0.43). In the entire group. MOPP chemotherapy (2 or 3 cycles) was followed by mantle irradiation of 40-45 Grays over 26 to 30 days and paraaortic splenic pedicle irradiation of 30 Grays over 19 days. Complete remission (CR) was achieved in 26 of 30 patients (87%). The remaining 4 patients who failed to respond to initial chemotherapy received salvage chemotherapy and/or autologous bone marrow transplantation (ABMT). With a median follow-up time of 72 months (range 11 to 131 months), overall survival and disease free survival (DFS) were 86% and 78% respectively at 10 years. Five relapses were observed after 10 to 63 months of CR. Among five patients (3 relapsing and 2 failing to respond) who received intensive chemotherapy and/or ABMT, complete remission was obtained in 4 (80%). Univariate analysis of different risk factors such as age, systemic symptoms, E-lesions, histological subtype, sex, MTR and response to initial chemotherapy indicated that early response to chemotherapy was the only significant factor influencing overall survival (p < 0.001). Intensive chemotherapy with ABMT is suggested for patients failing to respond to initial chemotherapy or relapsing after combined modality treatment. PMID- 8414966 TI - Founder effect in beta-thalassaemia in Portneuf, Quebec. AB - The genealogies of seven individuals with beta-thalassaemia minor carrying the (beta+IVS-1, nt110) mutation were reconstructed to determine the origin of the mutation in the French Canadian population of Portneuf County. A set of 55 ancestors common to all seven carriers was defined. These founders included 38 born in Europe of whom 34 came from 13 different regions of France and in particular from Languedoc (2 ancestors), a French province along the Mediterranean sea in which the mutation is still present. Descendants of these two individuals settled in Portneuf County where the gene frequency increased due to a high level of endogamy. However, present results cannot exclude a possibility that the (beta+IVS-1, nt110) mutation was introduced into the French Canadian population by settlers originating from a non-malarial region of France. The beta-thalassaemia gene has since spread from Portneuf County over the last century. PMID- 8414967 TI - Residual mediastinal mass in malignant lymphoma: value of magnetic resonance imaging and gallium scan. AB - Following treatment for mediastinal lymphoma, residual masses are defined as a mass greater than 2 cm observed on the CT scan in the absence of other evolutive signs of lymphoma. In this study, we examined 55 patients with residual mediastinal mass after optimal therapy, using gallium scan (37 cases) or magnetic resonance imaging (MRI, 44 cases). The group comprised 41 subjects with Hodgkin's disease and 14 with non Hodgkin's lymphoma, stages I and II (32 cases) and stages III and IV (23 cases); 35 subjects (64%) having bulky mediastinal involvement at diagnosis. A negative gallium scan or fibrotic signal on MRI was correlated with complete remission in 48 cases (87%). In seven of these 48 patients, MRI was not conclusive with a high signal of indeterminate tissue following radiotherapy, but four of the seven had fibrotic tissue on biopsy and none relapsed. Positive gallium scan was observed in six cases. In conclusion, we suggest performing a gallium scan at the end of induction chemotherapy and when it is negative, treatment may be continued without surgical biopsy or salvage therapy. MRI is of value when it shows fibrotic tissue but can be inconclusive. PMID- 8414968 TI - L-DNAs as potential antimessenger oligonucleotides: a reassessment. AB - Unnatural L-2'-deoxyribonucleosides L-T, L-dC, L-dA and L-dG were prepared from L arabinose and assembled, by solution or solid phase synthesis, to give L oligonucleotides (L-DNAs), which contain all four natural bases. The affinity of these modified oligomers for complementary D-ribo- and D-deoxyribo-oligomers was studied with NMR, UV and CD spectroscopies and mobility shift assay on native PAGE. All experimental results indicate that L-DNAs do not, in general, recognize single-stranded, natural DNA and RNA. Hence, contrary to previous suggestions, it is not possible to envisage their use as wide scope antimessenger agents in the selective control of gene expression. PMID- 8414969 TI - In vitro phosphorylation studies of a conserved region of the transcription factor ATF1. AB - A large family of mammalian transcription factors including multiple variants of CREB, CREM and ATF1 have been implicated in signal transduction by cAMP and other cellular pathways. Although the roles of some members of the family have been characterised the function of ATF1 is poorly understood. We have identified one or more key serine residues that are required for a phosphorylation-induced conformational change in ATF1. The critical serines map to a putative transcriptional activation domain of ATF1 and affect the stability of ATF1 DNA binding. Intriguingly phosphorylation is modulated by ATF1 homodimerization and by ATF1 binding to DNA. One of the key serine residues required for ATF1 phosphorylation is not conserved in CREB and CREM suggesting that it is likely to determine some specialised function of ATF1. PMID- 8414970 TI - A molecular basis for human hypersensitivity to aminoglycoside antibiotics. AB - We have investigated the distribution of mitochondrial DNA polymorphisms in a rare maternally transmitted genetic trait that causes hypersensitivity to aminoglycoside antibiotics, in the hope that a characterization of its molecular basis might provide a molecular and cellular understanding of aminoglycoside induced deafness (AGD). Here we report that the frequency of a particular mitochondrial DNA polymorphism, 1555G, is associated nonrandomly with aminoglycoside-induced deafness in two Japanese pedigrees, bringing the frequency of this polymorphism to 5 occurrences in 5 pedigrees of AGD, and in 4 of 78 sporadic cases in which deafness was thought to be the result of aminoglycoside exposure; both frequencies are significantly different from the occurrence of this mutation in the hearing population, which was 0 in 414 individuals surveyed. The 1555G polymorphism occurred in none of 34 aminoglycoside-resistant individuals. We propose a specific molecular mechanism for aminoglycoside hypersensitivity in individuals carrying the 1555G polymorphism, based on the three-dimensional structure of the ribosome, in which the 1555G polymorphism favors aminoglycoside binding sterically, by increasing access to the the ribosome cleft. PMID- 8414971 TI - A TBP-containing multiprotein complex (TIF-IB) mediates transcription specificity of murine RNA polymerase I. AB - TIF-IB is a transcription factor which interacts with the mouse ribosomal gene promoter and nucleates the formation of an initiation complex containing RNA polymerase I (Pol I). We have purified this factor to near homogeneity and demonstrate that TIF-IB is a large complex (< 200 kDa) which contains several polypeptides. One of the subunits present in this protein complex is the TATA binding protein (TBP) as revealed by copurification of TIF-IB activity and TBP over different chromatographic steps including immunoaffinity purification. In addition to TBP, three tightly associated proteins (TAFs-I) with apparent molecular weights of 95, 68, and 48 kDa are contained in this multimeric complex. This subunit composition is similar--but not identical--to the analogous human factor SL1. Depletion of TBP from TIF-IB-containing fractions by immunoprecipitation eliminates TIF-IB activity. Neither TBP alone nor fractions containing other TBP complexes are capable of substituting for TIF-IB activity. Therefore, TIF-IB is a unique complex with Pol I-specific TAFs distinct from other TBP-containing complexes. The identification of TBP as an integral part of the murine rDNA promoter-specific transcription initiation factor extends the previously noted similarity of transcriptional initiation by the three nuclear RNA polymerases and underscores the importance of TAFs in determining promoter specificity. PMID- 8414972 TI - A damage-recognition protein which binds to DNA containing interstrand cross links is absent or defective in Fanconi anemia, complementation group A, cells. AB - A DNA binding protein with specificity for DNA containing interstrand cross-links induced by 4,5',8-trimethylpsoralen (TMP) plus long wavelength ultraviolet (UVA) light has been identified in normal human chromatin. Protein binding to DNA was determined using a gel mobility shift assay and an oligonucleotide containing a hot spot for formation of psoralen interstrand cross-links. Specificity of the damage-recognition protein for cross-links was demonstrated both by a positive correlation between level of cross-link formation in DNA and extent of protein binding and by effective competition by treated but not undamaged DNA for the binding protein. Chromatin protein extracts from cells from individuals with the genetic disorder, Fanconi anemia, complementation group A (FA-A), which have decreased ability to repair damage produced by TMP plus UVA light, failed to show any protein binding to TMP plus UVA treated DNA. We have previously shown that these chromatin protein extracts contain a DNA endonuclease complex, pI 4.6, which specifically recognizes and incises DNA containing interstrand cross-links and which in FA-A cells is defective in its ability to incise this damaged DNA (Lambert et al. (1992) Mutation Res., 273, 57-71). Together, these findings suggest that the DNA binding protein identified is involved in recognition and repair of DNA interstrand cross-links. PMID- 8414973 TI - Mutagenesis analysis of a hepatitis delta virus genomic ribozyme. AB - We conducted extensive mutagenesis analysis on a hepatitis delta virus (HDV) genomic ribozyme to study the sequence specificity of certain region and to derive the secondary structure associated with the catalytic core. The results confirmed that the autocatalytic domain of HDV genomic RNA contained four base pairing regions as predicted in the 'pseudo-knot' model [Perrotta & Been (1990) Nature 350, 434-436]. The size and sequence of one of the base-pairing regions, i. e. stem-and-loop, could be flexible. Helix 3 and the first basepair of helix 1 required specific sequence to retain self-cleavage activity. The structural requirement of helix 2 was less stringent than the other base-pairing regions. Moreover, the size of helix 1 affected self-cleavage whereas the length of hinge could be variable even though the first three residues of hinge had stringent sequence requirement. PMID- 8414974 TI - Mutations dissociating the inhibitory activity of the pokeweed antiviral protein on eukaryote translation and Escherichia coli growth. AB - The pokeweed antiviral protein is a ribosome inactivating protein acting on eukaryotic as well as on prokaryotic ribosomes thus is toxic for both cell types. Using the PCR technique to clone the PAP open reading frame, we characterized two cDNAs coding for proteins inhibiting eukaryotic translation process and which are not toxic for Escherichia coli, unlike the wild type protein. The sequence of the two cDNAs showed that the proteins contain only one and two point mutations. This result suggest that the wild type amino acids in the mutated positions participate in the prokaryotic ribosome recognition. These mutants might be useful for the construction of immunotoxins containing the pokeweed antiviral protein as toxin. PMID- 8414975 TI - DNA fragmentation during apoptosis is caused by frequent single-strand cuts. AB - One of the hallmarks of apoptosis is the digestion of genomic DNA by an endonuclease, generating a ladder of small fragments of double-stranded DNA. We have examined the nature of the DNA breaks produced in mouse thymocytes triggered to undergo apoptosis by steroids or by stimulation of the T cell receptor. Whereas the typical ladder pattern of oligonucleosomal fragments was observed after agarose gel electrophoresis, numerous single-strand cuts were detected after electrophoresis under denaturing conditions. Single-strand nicks were found to be very frequent in the internucleosomal regions, but also to occur in the core particle-associated DNA. An identical pattern of single-strand nicks was obtained when chromatin DNA was exposed to the single-strand cleaving deoxyribonuclease I. The nicked DNA fragments, extracted from apoptotic thymocytes, were sensitive to the action of S1-nuclease. We propose that DNA fragmentation induced during apoptosis is not due to a double-strand cutting enzyme as previously postulated, but rather is the result of single-strand breaks. This ensures the dissociation of the DNA molecule at sites where cuts are found within close proximity. PMID- 8414976 TI - Increase in the basal transcriptional activity of the human foamy virus internal promoter by the homologous long terminal repeat promoter in cis. AB - The human foamy or spumaretrovirus HFV is a complex and exogenous retrovirus that encodes several bel genes besides the three classical retroviral genes gag, pol, and env. HFV was recently reported to contain two functionally active promoters that are both strongly trans-activated by the HFV trans-activator protein Bel 1. The occurrence of a second internal cap site underscores the complexity of the HFV genome. We have analysed whether there is interference between the HFV long terminal repeat promoter and the internal promoter located in the 3' end of env upstream of the bel genes. Recombinant clones were constructed that carry two different indicator genes, one under the control of the U3 promoter, the other under the control of the internal promoter. The portion of the basal transcriptional activity of the internal promoter that is not trans-activated by Bel 1 was increased two- to threefold in the presence of the long terminal repeat promoter. The rate of trans-activation by Bel 1 of both HFV promoters was not altered in these constructs. PMID- 8414977 TI - Roles of novobiocin-sensitive topoisomerases in chloroplast DNA replication in Chlamydomonas reinhardtii. AB - We have examined DNA replication in Chlamydomonas reinhardtii chloroplasts in vivo when chloroplast type II topoisomerases are inactivated with sublethal doses of novobiocin. DNA replication is at first inhibited under these conditions. However, after a delay of several hours, chloroplast chromosomes initiate a novobiocin-insensitive mode of DNA replication. This replication starts preferentially near a hotspot of recombination in the large inverted repeats, instead of from the normal chloroplast origins, oriA and oriB. It replicates one, but not the other single-copy region of the chloroplast chromosome. We speculate that novobiocin-insensitive DNA replication in chloroplasts requires recombination in this preferred initiation region. PMID- 8414978 TI - Stability, structure and complexity of yeast chromosome III. AB - The complete sequence of yeast chromosome III provides a model for studies relating DNA sequence and structure at different levels of organisation in eukaryotic chromosomes. DNA helical stability, intrinsic curvature and sequence complexity have been calculated for the complete chromosome. These features are compartmentalised at different levels of organisation. Compartmentalisation of thermal stability is observed from the level delineating coding/non-coding sequences, to higher levels of organisation which correspond to regions varying in G + C content. The three-dimensional path reveals a symmetrical structure for the chromosome, with a densely packed central region and more diffuse and linear subtelomeric regions. This interspersion of regions of high and low curvature is reflected at lower levels of organisation. Complexity of n-tuplets (n = 1 to 6) also reveals compartmentalisation of the chromosome at different levels of organisation, in many cases corresponding to the structural features. DNA stability, conformation and complexity delineate telomeres, centromere, autonomous replication sequences (ARS), transposition hotspots, recombination hotspots and the mating-type loci. PMID- 8414979 TI - How much hydration is necessary for the stabilisation of DNA-duplex? AB - A combination of NOESY and ROESY experiments show that the higher stabilities (T(m)) of phenazine tethered matched (2) and G-A mismatched (4) DNA duplexes are due to the decrease of the exchange-rate (i.e. increase of the life-time) of the imino-protons and the reduced water activity in their minor grooves compared to their non-tethered counterparts (1) and (3). PMID- 8414980 TI - Organization, inducible-expression and chromosome localization of the human HMG I(Y) nonhistone protein gene. AB - Members of the HMG-I(Y) family of mammalian nonhistone proteins are of importance because they have been demonstrated to bind specifically to the minor groove of A.T-rich sequences both in vitro and in vivo and to function as gene transcriptional regulatory proteins in vivo. Here we report the cloning, sequencing, characterization and chromosomal localization of the human HMG-I(Y) gene. The gene has several potential promoter/enhancer regions, a number of different transcription start sites and numerous alternatively spliced exons making it one of the most complex nonhistone chromatin protein-encoding genes so far reported. The putative promoter/enhancer regions each contain a number of conserved nucleotide sequences for potential binding of inducible regulatory transcription factors. Consistent with the presence of these conserved sequences, we found that transcription of the HMG-I(Y) gene is inducible in human lymphoid cells by factors such as phorbol esters and calcium ionophores. Detailed sequence analysis confirms our earlier suggestion that alternative splicing of precursor mRNAs gives rise to the major HMG-I and HMG-Y isoform proteins found in human cells. Furthermore, the gene's exon-intron arrangement fully accounts for all of the previously cloned human HMG-I(Y) cDNAs (1,2). Also of considerable interest is the fact that each of the three different DNA-binding domain peptides present in an individual HMG-I(Y) protein is coded for by sequences present on separate exons thus potentially allowing for exon 'shuffling' of these functional domains during evolution. And, finally, we localized the gene to the short arm of chromosome 6 (6p) in a region that is known to be involved in rearrangements, translocations and other abnormalities correlated with a number of human cancers. PMID- 8414981 TI - Influence of the three nucleotides upstream of the initiation codon on expression of the Escherichia coli lacZ gene in Saccharomyces cerevisiae. AB - By introducing synthetic oligonucleotides into a lacZ-yeast expression vector a set of 47 plasmids (out of 64 possible) was generated, differing only in the three bases immediately upstream of the AUG initiation codon of the Escherichia coli lacZ gene. Expression of the beta-galactosidase fusion protein encoded by the different plasmids was determined in Saccharomyces cerevisiae by immunogel electrophoresis. Among the clones tested we found a factor 3 difference in expression. A slight nucleotide preference was found in positions -3(A > G > C = U) and -2 (G > C = U > A). The choice of the nucleotide at position -1 immediately 5' of the AUG did not effect translation efficiency. Increasing homology to the yeast consensus sequence (AAAAAAAUGUCU) was not concomitant with an increased translation efficiency. Our results indicate that the choice of nucleotides immediately preceding the initiation codon in yeast does not dramatically influence translation efficiency, as in prokaryotes or higher eukaryotes. PMID- 8414982 TI - Identification of differentially expressed mRNA species by an improved display technique (DDRT-PCR). AB - We have significantly improved a method originally developed by Liang and Pardee [Science 257 (1992) 967-971] to display a broad spectrum of expressed genes and to detect differences in expression between different cell types. We have analysed various aspects of the technique and have modified it for both, the application to fast and efficient identification of genes and the use with automatic analysis systems. Based on the mathematical background we have devised the appropriate number of optimal PCR primers. We have also introduced nondenaturating gels for separating double stranded fragments as single bands. By applying the method to regenerating mouse liver, we have identified, out of a total of 38,000 bands, about 70 fragments where the expression of the corresponding genes seems to be differentially regulated at different time points. Application of the method to an automatic DNA sequencer was successfully done. Thus, we have confirmed the usefulness and increased the power of the RNA display technique, which we named differential display reverse transcription PCR (DDRT-PCR), and have extended the range of its application. PMID- 8414983 TI - (+)-CC-1065 as a structural probe of Mu transposase-induced bending of DNA: overcoming limitations of hydroxyl-radical footprinting. AB - Phage Mu transposase (A-protein) is primarily responsible for transposition of the Mu genome. The protein binds to six att sites, three at each end of Mu DNA. At most att sites interaction of a protein monomer with DNA is seen to occur over three minor and two consecutive major grooves and to result in bending up to about 90 degrees. To probe the directionality and locus of these A-protein induced bends, we have used the antitumor antibiotic (+)-CC-1065 as a structural probe. As a consequence of binding within the minor groove, (+)-CC-1065 is able to alkylate N3 of adenine in a sequence selective manner. This selectivity is partially determined by conformational flexibility of the DNA sequence, and the covalent adduct has a bent DNA structure in which narrowing of the minor groove has occurred. Using this drug in experiments in which either gel retardation or DNA strand breakage are used to monitor the stability of the A-protein--DNA complex or the (+)-CC-1065 alkylation sites on DNA (att site L3), we have demonstrated that of the three minor grooves implicated in the interaction with A protein, the peripheral two are 'open' or accessible to drug bonding following protein binding. These drug-bonding sites very likely represent binding at at least two A-protein-induced bending sites. Significantly, the locus of bending at these sites is spaced approximately two helical turns apart, and the bending is proposed to occur by narrowing of the minor groove of DNA. The intervening minor groove between these two peripheral sites is protected from (+)-CC-1065 alkylation. The results are discussed in reference to a proposed model for overall DNA bending in the A-protein att L3 site complex. This study illustrates the utility of (+)-CC-1065 as a probe for protein-induced bending of DNA, as well as for interactions of minor groove DNA bending proteins with DNA which may be masked in hydroxyl radical footprinting experiments. PMID- 8414984 TI - Direct estimation of base-pair exchange kinetics in oligo-DNA by a combination of NOESY and ROESY experiments. AB - A new method for the determination of the kinetics of exchange of the imino protons of DNA duplex is reported using a combination NOESY and ROESY experiments at short mixing times (< or = 20 ms). These results have been compared with the commonly used longitudinal relaxation approach through the T1 measurement. To calculate kex and pi ex by ROESY-NOESY experiment, the volume of the cross-peaks between imino protons and water in the NOESY and ROESY spectra have been measured separately from the magnetization term. This work shows that the present approach for the measurement of the kinetics of slow exchanging imino protons of DNA duplex is comparable to the saturation recovery experiment in which the exchange rate can be accelerated by the addition of a base catalyst. The present ROESY NOESY approach has been found to be particularly useful and reasonably accurate for the measurement of exchange kinetics of both the fast- and slow-exchanging imino protons in DNA duplex both under non-physiological and physiological condition where the saturation recovery method can not be used. PMID- 8414985 TI - DNA replication facilitates the action of transcriptional enhancers in transient expression assays. AB - We demonstrate a general role for DNA replication in the activation of gene transcription in transient transfection assays. The effect is observed for a wide range of genes and cell types, transfected by a number of protocols and is independent of increased template copy number. Replication does not stimulate transcription driven by proximal promoter elements alone but requires a functional enhancer element. This synergy between an active replication origin and an enhancer is not confined to elements from viruses such as SV40, which undergo an early to late switch in gene expression that is tightly coupled to replication, since the enhancer-containing long terminal repeats from retroviruses are strongly stimulated by replication. Furthermore, synthetic enhancers consisting of multimerised binding sites for one or two factors are also subject to replication-activation. The diversity of synthetic and natural enhancers used in this study suggests that replication and transcription do not share a common protein factor. We propose that replication leads to chromatin modifications that facilitate enhancer action. PMID- 8414986 TI - Frequent amplification of a short chain dehydrogenase gene as part of circular and linear amplicons in methotrexate resistant Leishmania. AB - The H locus of Leishmania codes for a short chain dehydrogenase gene (ltdh) that is involved in antifolate resistance. Leishmania tarentolae cells, selected in a step by step fashion for resistance to the antifolate methotrexate (MTX), frequently amplified ltdh in response to drug selection. Both circular and linear extrachromosomal amplicons were generated de novo from the chromosomal H locus and several contained inverted duplications. At least four different rearrangement points were used during the formation of amplicons, with one of them used preferentially. All mutants highly resistant to MTX, whether or not they have the H locus amplified, showed a decreased steady-state accumulation of MTX. Nevertheless, two types of transport mutants were clearly discernable. In the first type, accumulation was reduced four to five-fold, whereas in the second class of mutants, accumulation was reduced more than 50-fold. The ltdh gene was amplified in all the mutants with the transport mutation of the first type, but not in all the mutants with a more pronounced decrease in the steady-state accumulation of MTX. Both types of transport mutation, leading to the reduction in MTX accumulation, arose early during the selection process and were stable even when cells were grown in absence of the drug for prolonged period. PMID- 8414987 TI - Trypanosoma brucei minicircles encode multiple guide RNAs which can direct editing of extensively overlapping sequences. AB - Small guide RNAs (gRNAs) may direct RNA editing in kinetoplastid mitochondria. We have characterized multiple gRNA genes from Trypanosoma brucei (EATRO 164), that can specify up to 30% of the editing of the COIII, ND7, ND8, and A6 mRNAs and we have also found that the non-translated region of edited COIII mRNA of strain (EATRO 164) differs from that of another strain. Several of the gRNAs specify overlapping regions of the same mRNA often specifying sequence beyond that required for an anchor duplex with the next gRNA. Some gRNAs have different sequence but specify identical editing of the same region of mRNA. These data indicate a complex gRNA population and consequent complex pattern of editing in T. brucei. PMID- 8414988 TI - An upstream promoter element of the Acanthamoeba castellanii TBP gene binds a DNA sequence specific transcription activating protein, TPBF. AB - We have characterized a positive-acting element in the upstream portion of the Acanthamoeba TBP gene promoter. The 27 bp element (TPE), located within the promoter between -97 and -70, stimulates transcription in an orientation independent fashion and tolerates modest changes in its distance from the TATA box. The TPE does not, however, function synergistically nor when positioned 3000 bp 5' or 260 base pairs 3' of the transcription start site. The TPE binds a DNA sequence-dependent factor, TPBF, which we have partly purified. TPBF was characterized using in vitro transcription, DNase I footprinting, methylation interference and electrophoretic mobility shift assays. TPBF does not have a counterpart in HeLa cells, but nonetheless strongly stimulates transcription of the Acanthamoeba TBP gene in mammalian extracts. Our results also suggest that there are additional positively and negatively acting elements within the TBP gene promoter, for which a model is presented. PMID- 8414989 TI - Use of PCR primers containing a 3'-terminal ribose residue to prevent cross contamination of amplified sequences. AB - Cross-contamination with previously amplified products poses a serious limitation in the use of PCR for clinical testing and in certain research applications as well. In the present study we report the use of novel primers containing a 3' terminal ribose residue to circumvent this problem. Extension of the primer by Taq DNA polymerase generates a cleavable ribonucleotide linkage within the amplified product. Cleavage of the primer by base or with a ribonuclease interferes with further replication of the product should carry over to another sample occur. Primers terminating in any of the 4 ribose residues function equally well as all DNA primers. Taq DNA polymerase is thus able to both efficiently extend and copy the single ribose residue. In translating from all DNA primers to ones containing a 3'-ribose residue no modification of the PCR protocol is required. The products formed can be used in all applications of the PCR. Since neither the original sample DNA, the primers or the extension products are modified by base or ribonuclease treatment both pre- and post-amplification sterilization can be carried out. Pre-amplification treatment with RNase A can yield as high as 10(4)-fold sterilization. Under these conditions the addition of beta-mercaptoethanol or other sulfhydryl reducing agent is necessary to inactivate the enzyme during thermocycling. Post-amplification treatment with NaOH readily yields at least 10(6)-fold sterilization. This alone is sufficient for most, if not all, applications of PCR. It is especially useful for quantitative RT-PCR, since the original target RNA sequence, which may be present in high copy numbers, is also destroyed. PMID- 8414990 TI - Cloning and characterization of the C. elegans histidyl-tRNA synthetase gene. AB - In this paper, we report the cloning and sequencing of the C. elegans histidyl tRNA synthetase gene. The complete genomic sequence, and most of the cDNA sequence, of this gene is now determined. The gene size including flanking and coding regions is 2230 nucleotides long. Three small introns (45-50 bp long) are found to interrupt the open reading frame. The open reading frame translates to 523 amino acids. This putative protein sequence shows extensive homology with the human and yeast histidyl-tRNA the histidyl-tRNA synthetase gene is a single copy gene. Hence, it is very likely that it encodes both the cytoplasmic and the mitochondrial histidyl-tRNA synthetases. It is likely to be trans-spliced since it contains a trans-splice site in its 5' untranslated region. PMID- 8414991 TI - Dimerization of leucine zippers analyzed by random selection. AB - The leucine zipper is a coiled coil that mediates specific dimerization of bZIP DNA-binding domains. A hydrophobic spine involving the conserved leucines runs down the coiled-coil and is thought to stabilize the dimer. We used the method of random selection to further define the primary sequence requirements for homodimer formation and heterodimer formation with Fos. When positions on either side of the hydrophobic spine of GCN4 are diversified to include the corresponding residues of Jun, a large percentage of the resulting sequences form homodimers, and a large percentage form heterodimers with Fos. Basic residues were preferred, but not essential, at position e of zippers which heterodimerize with Fos. When random sequences containing 5 heptad repeat of leucines are subject to a selection for homodimer formation, a diverse set of sequences is isolated. Certain residues are preferred at each position in the heptad repeat, although no essential primary sequence determinants could be identified. No pair of residues not involving the conserved leucines could be identified which strongly promotes homodimerization. These results suggest that factors determining leucine zipper dimerization are complex, with numerous interactions contributing to the association. PMID- 8414992 TI - rseB, a chromosomal locus that affects the stability of a temperature-specific surface protein mRNA in Tetrahymena thermophila. AB - In Tetrahymena thermophila, the expression of a temperature-specific surface protein known as SerH3 is primarily controlled by a temperature-dependent change in the stability of the mRNA that encodes this protein. At 30 degrees C the SerH3 mRNA displays a half-life of 60 minutes while at 40 degrees C the half-life decreases to only 3 minutes. We used a Tetrahymena mutant cell line (rseB) defective in expression of SerH3 at 30 degrees C to explore the mechanisms involved in temperature-dependent mRNA stability. The results of in vitro nuclear run-off assays and Northern and slot blot analysis of cytoplasmic and nuclear RNAs show that the rseB locus encodes a temperature-sensitive product that has no effect on SerH3 gene transcription or the steady-state levels of SerH3 nuclear RNA. However, the product of this locus does have a dramatic effect on cytoplasmic levels of the SerH3 mRNA at 30 degrees C, indicating that SerH3 gene expression is affected post-transcriptionally within the cytoplasm. To explore the possibility that the rseB locus controls SerH3 mRNA stability we developed an in vitro mRNA decay assay. This assay successfully duplicates the differential decay of the SerH3 mRNA observed in wild-type cells grown at different temperatures. The apparent half-life of the SerH3 mRNA in cytoplasmic extracts derived from cells grown at 30 degrees C is approximately 45 minutes while in cytoplasmic extracts derived from cells grown at 40 degrees C it is only 6 minutes. When similar experiments are performed using extracts prepared from the Tetrahymena rseB cell line, we find that the SerH3 mRNA is only stable in extract prepared from cells grown under conditions in which the mRNA accumulates to detectable levels in the cytoplasm. These results indicate that the product of the rseB locus is a trans-acting cytoplasmic factor that exerts its effect on SerH3 gene expression by regulating SerH3 mRNA stability. PMID- 8414993 TI - A topoisomerase II-like protein is part of an inducible DNA-binding protein complex that binds 5' of an immunoglobulin promoter. AB - We previously identified a B cell-specific protein-DNA complex 5' of an immunoglobulin mu heavy chain promoter. The sequences to which this protein complex bound were required for induction of immunoglobulin mRNA levels with interleukin-5 + antigen. Further studies identified a second sequence 5' of these regulatory sequences that bound to both the nuclear matrix and to a similar interleukin-5 + antigen inducible protein complex. Therefore, we sought to identify the putative regulatory proteins that comprised this DNA-binding complex. In this study, we have used anti-topoisomerase II antibodies to demonstrate that one of the proteins found in the interleukin-5 + antigen inducible complexes is serologically related to topoisomerase II. To our knowledge, this is the first report where a topoisomerase II related protein participates in an inducible mobility shifted protein-DNA complex. These data suggest a model in which the enhanced immunoglobulin gene transcription observed after treatment with interleukin-5 + antigen might be explained by the induction of a protein complex that acts to relieve torsional stress along the gene. PMID- 8414994 TI - dHMG-Z, a second HMG-1-related protein in Drosophila melanogaster. AB - We report the identification of dHMG-Z, a gene related to dHMG-D and encoding a second invertebrate homologue of HMG 1 protein. The encoded proteins is 65% identical to dHMG-D protein, and also contains a single HMG-box as the DNA recognition motif. Analogous to dHMG-D, two transcripts are observed for dHMG-Z which are differentially regulated, and are the product of zygotic transcription unlike the dHMG-D transcripts which arise from both maternal and zygotic transcription. The genes for dHMG-D and dHMG-Z are located on adjacent loci in the genome and each contains two introns. The position of the second intron in the coding region is conserved between the two genes suggesting a common origin via gene duplication. PMID- 8414995 TI - The tyrosine phosphatase cdc25 selectively inhibits transcription of the Xenopus oocyte-type tRNAtyrC gene. AB - The Xenopus tyrosine tRNAtyrC (TyrC) genes are developmentally regulated. These multicopy genes are expressed in early oocytes and inactivated as oocytes reach maturity. As shown here, this developmental regulation can be reproduced in vitro in extracts of early and late stage oocytes: the TyrC gene is transcribed in early oocyte extracts but is virtually inactive in mature oocyte extracts. The inability to transcribe the TyrC gene is not due to the lack of functional pol III transcriptional components, since the somatic-type TyrD gene is fully active in mature oocyte extracts. Instead, the loss of TyrC transcription appears to be due to a change in the template specificity of transcription factor TFIIIC: addition of TFIIIC from immature extracts restores TyrC transcription in mature extracts. In mixtures of immature and mature extracts, the transcriptional activity of the TyrC gene is reduced. The presence of sodium vanadate, an inhibitor of tyrosine phosphatases, increases the level of TyrC transcription in the extract mixtures. Also, cdc25 phosphatase treatment of immature extracts causes a decrease in TyrC transcription which is reversed by addition of exogenous TFIIIC. These findings indicate that changes in phosphorylation state alters the template specificity of TFIIIC leading to the selective inactivation of oocyte type TyrC genes. PMID- 8414996 TI - Autoregulation of GAL4 transcription is essential for rapid growth of Kluyveromyces lactis on lactose and galactose. AB - Transcriptional induction of genes in the lactose-galactose regulon of the yeast Kluyveromyces lactis requires the GAL4 transcription activator protein. Previous data indicated that the concentration of GAL4 was tightly regulated under basal, inducing, and glucose repressing conditions but the mechanisms were unknown. In this paper we demonstrate that transcription of the GAL4 gene (KI-GAL4) increases 3- to 4-fold during induction of the regulon. This increase requires a KI-GAL4 binding site, UASG, in front of the KI-GAL4 gene, indicating that the KI-GAL4 protein autoregulates transcription of its own gene. Our data demonstrate that the autoregulatory circuit is essential for full induction of the lactose galactose regulon and, hence, for rapid growth on lactose or galactose. Other data indicate that basal transcription of the KI-GAL4 gene is governed by unidentified promoter elements. The existence of the autoregulatory circuit reveals an important difference between the lactose-galactose regulon and its homologue in Saccharomyces cerevisiae, the melibiose-galactose regulon. This difference may have evolved in response to different selective pressures encountered by the two organisms. PMID- 8414997 TI - Quantitative evaluation of intracellular sense: antisense RNA hybrid duplexes. AB - Previous studies have demonstrated that for an antisense RNA to be effective in attenuating gene expression, a large but indeterminate excess of antisense RNA is required. To quantitatively evaluate RNA hybrid duplex formation, expression vectors containing antisense dihydrofolate reductase (DHFR) cDNAs were transfected into KB and KB-1BT (a DHFR overexpressing variant) cells and transfectants expressing antisense transcripts of exon 1 through intron I (ex1-I) or exons 1 through 4 (ex1-4) were analyzed for hybrid duplex formation. Stable duplexes were detectable in KB-1BT but not in KB cells. Approximately 5-9% of antisense ex1-I RNA and 20-37% of antisense ex1-4 RNA were found in duplexes. The amount of each hybrid duplex RNA was found to be a linear function of intracellular single-stranded antisense RNA levels and a hybrid index, Hs:as, was devised to describe this relationship. Based upon the value of Hs:as for each antisense RNA:mRNA duplex, it is calculated that an approximate 2,800- and 600 fold excess of ex1-I and ex1-4 antisense RNA are respectively required for 50% of DHFR mRNA to be present in duplexes. Results support the hypothesis that intracellular sense:antisense RNA hybrid duplex formation is inefficient and dependent upon the levels, lengths and possibly the structures of the RNAs involved. PMID- 8414998 TI - Sequence of the mitochondrial control region, tRNA(Thr), tRNA(Pro) and tRNA(Phe) genes from the black rhinoceros, Diceros bicornis. PMID- 8414999 TI - Genomic sequence of mitochondrial genes coding for ATPase subunit 6 and small subunit ribosomal RNA from Penicillium chrysogenum: a key for molecular systematics on fungi. PMID- 8415000 TI - A cDNA sequence of human ribosomal protein, homologue of yeast S28. PMID- 8415001 TI - A new human ribosomal protein sequence, homologue of rat L9. PMID- 8415002 TI - Tsp45I, a new thermostable site-specific endonuclease that cleaves the recognition sequence 5'-decreases GTSAC-3'. PMID- 8415003 TI - Identification of two human homologues to Drosophila SOS (son of sevenless) localized on two different chromosomes. PMID- 8415004 TI - Primary sequence of the mouse ribosomal protein L37a. PMID- 8415005 TI - Nucleotide sequence of a portion of the 26S rDNA of Acanthamoeba. PMID- 8415006 TI - Predicted editing of additional transfer RNAs in Acanthamoeba castellanii mitochondria. PMID- 8415007 TI - Nucleotide sequence of a cDNA encoding an alternative form of LEF-1. PMID- 8415008 TI - Nucleotide sequence analysis of the A protein of the U1 small nuclear ribonucleoprotein particle: the murine protein contains a 5' amino-terminal tag. PMID- 8415009 TI - A family of nuclear homing endonucleases. PMID- 8415010 TI - Conserved sites in the 5'-3' exonuclease domain of Escherichia coli DNA polymerase. PMID- 8415011 TI - A simple mechanical procedure to produce encapsulated cells. PMID- 8415012 TI - Anhydrotetracycline, a novel effector for tetracycline controlled gene expression systems in eukaryotic cells. PMID- 8415013 TI - An efficient method to generate phosphatase insensitive 3' labelled DNA probes using Taq polymerase. PMID- 8415014 TI - High efficiency transformation of Saccharomyces cerevisiae by electroporation. PMID- 8415015 TI - Antibody detection of protein complexes bound to DNA. PMID- 8415016 TI - Simultaneous randomization of antibody CDRs by a synthetic ligase chain reaction strategy. PMID- 8415017 TI - Direct autoradiography after radio-ligand binding to identify mammalian cell clones expressing hormone receptor cDNA. PMID- 8415018 TI - cDNA selection from total yeast DNA containing YACs. PMID- 8415019 TI - Addition of a class IIS enzyme site in the mutagenic primer to improve two-step PCR-based targeted mutagenesis. PMID- 8415020 TI - Elongation factor Tu as a control gene for mRNA analysis of lung development and other differentiation and growth regulated systems. PMID- 8415021 TI - Incorporation of dITP or 7-deaza dGTP during PCR improves sequencing of the product. PMID- 8415022 TI - Effect of drugs on gene expression in mammalian cells: a highly efficient procedure to test large numbers of samples. PMID- 8415023 TI - Sequence and secondary structure of 5.8S rRNA in the tick, Ixodes scapularis. PMID- 8415024 TI - A novel method to stabilise antisense oligonucleotides against exonuclease degradation. PMID- 8415025 TI - The challenge of reform. PMID- 8415026 TI - Research! America: building public support for research. PMID- 8415027 TI - A creative clinical education model: three views. AB - To effectively educate nurses who are able to meet changing health care needs, new models for clinical experiences must be considered. Collaborative arrangements between practice and education are particularly useful. The preceptor approach is used widely for seasoned students in senior experiences nearing graduation. The "paired" model, a variation on the preceptor model, has worked well in one setting for beginning students. It needs to be tried in other settings with different faculty and perhaps in other types of courses, and it needs to be tested. Collaboration between education and practice not only benefits education, but lays the groundwork for shared scholarly endeavors. Ultimately these relationships have a positive impact on patient care. PMID- 8415028 TI - Posology errors by sophomore nursing students. PMID- 8415029 TI - The chemically dependent student nurse: guidelines for policy development. PMID- 8415030 TI - Nutrition in nursing curricula: recent developments and recommendations. AB - In summary, nursing and nutrition have matured as scholarly professional disciplines. Both have bodies of knowledge and professional practice arenas that are unique, yet interrelated. Professional maturity and social necessity are resulting in increased collaboration and cooperation between nurses and dietitians after a time of separation and lack of communication. The status of nutrition education for nurses at this time is complex. There is consensus that nurses have a role in the nutritional care of clients and that nutrition content is essential. But what that content should be has not been generally accepted. Consequently, questions related to what content should be emphasized, its proper placement in the curriculum, and the best way to teach nutrition have not been answered. The eight guidelines developed by Morse offer a basis for the development, implementation and evaluation of essential nutrition content in undergraduate nursing curricula. The SNE content used in conjunction with these guidelines can help direct the selection of specific content. The professional background of the educator who teaches nutrition in nursing curricula influences content. We believe that both nurses and dietitians must be actively involved in the nutrition education of nurses. Both professions have as part of their perspective the health of people. The focus of nursing has been described as "the study of caring in the human health experience." The focus of dietetics has been described as the study of the impact of food and nutrients on the health of people.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415031 TI - Factors and work settings that may influence nurse practitioner practice. PMID- 8415032 TI - A new agenda for nursing education. PMID- 8415033 TI - A new agenda for nursing education. PMID- 8415034 TI - A new agenda for nursing education. PMID- 8415035 TI - A new agenda for nursing education. PMID- 8415036 TI - What's in a name? PMID- 8415037 TI - What does the NCATS (Nursing Child Assessment Teaching Scale) measure? AB - The purpose of this study was to examine what aspects of the mother-child relationship are measured by the Nursing Child Assessment Teaching Scale (NCATS). A racially heterogeneous sample of 128 mothers completed questionnaires measuring maternal depression, parenting self-efficacy, knowledge of developmental and parenting principles, and perceived difficult toddler temperament. Mothers and children were also videotaped during home visits while completing two teaching tasks that were later scored using NCATS. NCATS Parent subscale scores were significantly related to maternal knowledge and education but unrelated to depression and self-efficacy. Child subscale scores were unrelated to all of the study variables, including perceived difficult temperament. Significant differences were noted among African-American, Hispanic, and white mothers. The findings suggest that the NCATS taps cognitive factors more reliably than affective factors underlying the mother-child relationship and the cognitive factors may be culturally biased. PMID- 8415038 TI - Further psychometric evaluation of the Spanish language health-promoting lifestyle profile. AB - The reliability and validity of the Spanish language version of the Health Promoting Lifestyle Profile (Spanish-HPLP) was evaluated for a predominantly Central American sample. A convenience sample (N = 106) completed the demographic sheet and Spanish-HPLP. Alpha coefficients were 0.94 for the total scale and ranged from 0.64 to 0.89 for the subscales. Significant Pearson correlations were found between Spanish-HPLP scores and the variables of age, education, income, length of residence in the United States, and perceived health status. Significant differences based on sex and marital status were noted. The Spanish HPLP was found to be reliable for this sample. Although content and construct validity were supported, the instrument is in need of further convergent, or criterion-related validation. PMID- 8415039 TI - Influence of functional, urological, and environmental characteristics on urinary incontinence in community-dwelling older women. AB - The purpose of this study was to explore the relationships of functional, urological, and environmental characteristics to the frequency of urinary incontinence in 131 community-dwelling older women. Subjects with detrusor instability with or without concomitant genuine stress incontinence had significantly more impaired physical functioning, slower gait speeds, smaller bladder capacities, and less ability to delay voiding than subjects with genuine stress incontinence alone. Age, distance to toilet used most, and toilet-gait speed explained 17% of the variance in incontinence severity. Younger age, slower mobility, and a shorter distance to reach the toilet were associated with a higher frequency of incontinence. Physical functional status and urological characteristics, including urodynamic diagnosis, did not predict incontinence severity. These findings confirm, in part, the commonly held assumption that mobility and the environment influence urinary incontinence. PMID- 8415040 TI - Theoretical and methodological differentiation of moderation and mediation. PMID- 8415041 TI - The applicability of sequential analysis to nursing research. PMID- 8415042 TI - Examining amount and pattern of change: comparing repeated measures ANOVA and individual regression analysis. PMID- 8415043 TI - Permuted block design for randomization in a nursing clinical trial. PMID- 8415044 TI - Systematic sampling: theory and clinical methods. PMID- 8415045 TI - To correlate or not to correlate: what is the question? PMID- 8415046 TI - Instrument format issues in assessing the elderly: the Iowa Self-Assessment Inventory. PMID- 8415047 TI - Self-as-instrument in qualitative research. PMID- 8415048 TI - Nonnutritive sucking: effects on crying and heart rate in intubated infants requiring assisted mechanical ventilation. PMID- 8415049 TI - Evaluating abstracts: preparing a research conference. AB - Refusal to recognize the impossibility of designing a fair scholarly screening system for selecting papers from abstracts results in the use of inappropriate criteria and checklists. Since no checklist can be used universally to identify the rigorous and significant research contributions from abstracts, we suggest that a more flexible and sensible method of screening presentations is to select the maximum number based on the time and space available for presentations, the program objectives, and content needs. PMID- 8415050 TI - Coding dummy variables and calculating the relative risk in a logistic regression. PMID- 8415051 TI - Keeping abreast of the literature electronically. PMID- 8415052 TI - Inventing nursing research. PMID- 8415054 TI - Are highly dependent clients put at risk by community care? PMID- 8415053 TI - Teaching and learning 1. (i) The learning experience. PMID- 8415055 TI - Fields of conflict. PMID- 8415056 TI - Trial after error. PMID- 8415057 TI - Market mayhem. PMID- 8415058 TI - Travel sickness. PMID- 8415059 TI - Patient knows best. PMID- 8415060 TI - Summertime blues. PMID- 8415061 TI - There's no place like home--early discharge. PMID- 8415062 TI - Is home best?--early discharge. PMID- 8415063 TI - Transplants--moral costs. PMID- 8415064 TI - Mental health--inside the walls. PMID- 8415065 TI - Complementary medicine: nutritional therapies. PMID- 8415066 TI - Cervical screening among Asian women. PMID- 8415068 TI - Open learning--a learning experience. PMID- 8415067 TI - IV heparin therapy and urinalysis. PMID- 8415069 TI - Cow's milk allergy. PMID- 8415070 TI - 1. Teaching and learning (ii). How adults learn. PMID- 8415071 TI - Culture clash. PMID- 8415072 TI - Tortuous treatment? PMID- 8415073 TI - The missing link. PMID- 8415074 TI - Identities paraded. PMID- 8415075 TI - Do the continental. PMID- 8415077 TI - All snakes and no ladders. PMID- 8415076 TI - It's a man's world. PMID- 8415078 TI - Poor relations. PMID- 8415079 TI - Sunshine after the rain. PMID- 8415080 TI - Speak up. PMID- 8415081 TI - Child health. Acting with restraint. PMID- 8415083 TI - A short-sighted policy. PMID- 8415082 TI - Comparing qualified nurse and auxiliary roles. AB - This paper describes the contribution of nursing auxiliaries (NAs) compared with qualified nurses (QNs) in terms of activities performed. It also examines how this was affected by three organisational types, namely primary, team and functional nursing. Twelve QNs and 12 NAs were chosen randomly in each organisational type. Data were collected using non-participant observation and a computerised event recorder. The most interesting findings were found across organisational type, with QNs and NAs within each type engaging in similar patterns of work. Both grades of nursing staff in primary wards spent a greater amount of time in direct patient care but less time in supplementary patient care and staff activities. Team and functional nursing staff spent more time with patients in domestic and administrative activities. The implications of these findings for the employment of NAs in care of the elderly wards are discussed. PMID- 8415084 TI - Inappropriate use of acute psychiatric beds. PMID- 8415085 TI - Travel precautions. PMID- 8415086 TI - Practice nurses. Training questions. PMID- 8415087 TI - Practice nurses. Partner potential. PMID- 8415088 TI - Palliative care. Age-related problems. PMID- 8415089 TI - Managing cancer pain. PMID- 8415090 TI - The learning context (i). Examining the learning context. PMID- 8415091 TI - Redesigning life. PMID- 8415093 TI - Putting pay to nurses' hopes. PMID- 8415092 TI - Theory in practice. PMID- 8415094 TI - Wintry prospects. PMID- 8415095 TI - Eye contact. PMID- 8415096 TI - Day-release cataracts. PMID- 8415097 TI - Time for action. PMID- 8415098 TI - Meeting a need. PMID- 8415100 TI - Kill or cure? PMID- 8415099 TI - Child health. A family affair. PMID- 8415101 TI - Back in control. PMID- 8415102 TI - Birth trauma. PMID- 8415103 TI - The appliance of science. PMID- 8415104 TI - Continence. Reformed role. PMID- 8415105 TI - Continence. Target practice. PMID- 8415106 TI - Continence. Island life. PMID- 8415107 TI - Continence. Action plans. PMID- 8415108 TI - The learning context (ii). Curriculum models. PMID- 8415109 TI - Cutting decision. Interview by Tricia Reid. PMID- 8415110 TI - Home rules. PMID- 8415112 TI - Whither supervision? PMID- 8415111 TI - Acute uncertainty. PMID- 8415113 TI - Making sense of Swan-Ganz monitoring. PMID- 8415114 TI - Heartfelt support. PMID- 8415115 TI - Recommended reading. PMID- 8415117 TI - Manual handling. Too heavy to handle? PMID- 8415116 TI - Hypnotherapy. PMID- 8415118 TI - Down with ivory towers. PMID- 8415119 TI - Community care. Outside risk. PMID- 8415120 TI - Computing. Achieving acceptance. PMID- 8415121 TI - Learning by numbers. PMID- 8415122 TI - Health visitors. Gaining ground. PMID- 8415123 TI - Health visitors. Worthwhile changes. PMID- 8415124 TI - Health visitors. Courses of action. PMID- 8415125 TI - Inhibition of tumor promoter-induced hydrogen peroxide formation in vitro and in vivo by genistein. AB - Here we report that genistein, a soybean isoflavone, strongly inhibits tumor promoter-induced H2O2 formation both in vivo and in vitro. Genistein suppressed H2O2 production by 12-O-tetradecanoylphorbol-13-acetate- (TPA) stimulated human polymorphonuclear leukocytes (PMNs) and HL-60 cells in a dose-dependent manner over the concentration range 1-150 microM. Human PMNs were more sensitive to the inhibitory effect of genistein than HL-60 cells (50% inhibitory concentration 14.8 and 30.2 microM, respectively). In addition, genistein moderately inhibited superoxide anion formation by HL-60 cells and scavenged exogenously added H2O2 under the same conditions as in cell culture. However, the H2O2-scavenging effect of genistein was about 50% lower than its inhibition of cell-derived H2O2 formation at all concentrations. In the CD-1 mouse skin model, genistein strongly inhibited TPA-induced oxidant formation, edema, and PMN infiltration in mouse skin. Inhibition of TPA-mediated H2O2 in vivo may result from decreased cell derived H2O2 formation, scavenging of H2O2 produced, and/or suppression of PMN infiltration into the dermis. The antioxidant properties of genistein may be responsible for its anticarcinogenic effects, and the dietary availability of genistein makes it a promising candidate for the prevention of human cancers. PMID- 8415126 TI - Correlations of colon cancer mortality with dietary factors, serum markers, and schistosomiasis in China. AB - To determine correlates of the geographic variation in colon cancer mortality within China, dietary variables, biochemical markers, and other factors from an ecological survey in 49 Chinese rural counties were examined. High consumption of animal foods, salt-preserved vegetables, and beer was associated with increased mortality of colon cancer, whereas the rates were significantly inversely related with intake of green vegetables. Serum levels of total cholesterol, urea nitrogen, and lipid peroxide were positively correlated with colon cancer mortality, after adjustment for each other and for other blood nutrients. No appreciable associations, however, were found between colon cancer and serum levels of beta-carotene, alpha-tocopherol, vitamin C, and selenium. In addition, prevalence of schistosomiasis was significantly correlated with increased colon cancer mortality. This ecological study indicates that observations from earlier analytic investigations in Western societies may apply to a Chinese rural population and suggests that schistosomiasis and dietary factors may contribute to the remarkable geographic variation of colon cancer in China. PMID- 8415127 TI - Intake of potentially anticarcinogenic flavonoids and their determinants in adults in The Netherlands. AB - Flavonoids are strong antioxidants that occur naturally in foods and can inhibit carcinogenesis in rodents. Accurate data on population-wide intakes of flavonoids are not available. Here, using data of the Dutch National Food Consumption Survey 1987-1988, we report the intake of the potentially anticarcinogenic flavonoids quercetin, kaempferol, myricetin, apigenin, and luteolin among 4,112 adults. The flavonoid content of vegetables, fruits, and beverages was determined by high performance liquid chromatography. In all subjects, average intake of all flavonoids combined was 23 mg/day. The most important flavonoid was the flavonol quercetin (mean intake 16 mg/day). The most important sources of flavonoids were tea (48% of total intake), onions (29%), and apples (7%). Flavonoid intake did not vary between seasons; it was not correlated with total energy intake (r = 0.001), and it was only weakly correlated with the intake of vitamin A (retinol equivalents, r = 0.14), dietary fiber (r = 0.21), and vitamin C (r = 0.26). Our use of new analytic technology suggests that in the past flavonoid intake has been overestimated fivefold. However, on a milligram-per-day basis, the intake of the antioxidant flavonoids still exceeded that of the antioxidants beta-carotene and vitamin E. Thus flavonoids represent an important source of antioxidants in the human diet. PMID- 8415128 TI - Increased levels of S-adenosylmethionine in the livers of rats fed various forms of selenium. AB - The levels of S-adenosylmethionine (SAM) were determined in the livers of male weanling rats fed six different forms of selenium for five weeks. The following forms of selenium were administrated in the diet in logarithmic doses: sodium selenite, sodium selenate, sodium selenide, selenomethionine, selenocystine, and selenium sulfide. An overall increase in hepatic SAM was observed, and all compounds resulted in at least one observation where SAM was significantly elevated (p < 0.01). No dose-response relationship was found to exist, however. A comparison of the relative toxicity of each of the selenicals was based on a dose of 10 ppm dietary selenium for each chemical form. The elevation of SAM resulting from the subchronic administration of selenium may be one mechanism involved in the well-known chemopreventive effects in experimental models. PMID- 8415129 TI - Effects of rice bran hemicellulose on 1,2-dimethylhydrazine-induced intestinal carcinogenesis in Fischer 344 rats. AB - The effect of rice bran hemicellulose (RBH) on 1,2-dimethylhydrazine- (DMH) induced intestinal carcinogenesis was studied in male Fischer 344 rats. Rats were fed a basal control diet or a diet containing 2% or 4% RBH at five weeks of age. At 6 weeks of age, all animals were given an intraperitoneal injection of DMH (20 mg/kg body wt) at weekly intervals for 20 weeks and autopsied 7 weeks after the last injection. The incidence of DMH-induced colon tumors was significantly lower in rats fed the 4% RBH diet than in rats fed the basal control diet (p < 0.05). The number of colon tumors per rat was also significantly lower in rats fed the 4% RBH diet than in rats fed the basal control diet (p < 0.05). The present study suggested that the water-soluble RBH played a preventive role in DMH-induced large bowel carcinogenesis in Fischer 344 rats. PMID- 8415130 TI - Undernutrition as a risk factor for cervical intraepithelial neoplasia: a case control analysis. AB - To evaluate the relationship between cervical intraepithelial neoplasia (CIN) and undernutrition, a pair-matched case-control study was conducted in a low-income urban population. As a broad measure of nutritional status, serum albumin, serum ferritin, hematocrit, percent desirable weight, and percent calories consumed as protein were examined. Cases (n = 102) had biopsy-confirmed CIN I, II, or III, and clinic controls (n = 102), matched on age, race, and clinic, had normal Pap smears. Survey-collected data and frozen serum were utilized to study the hypothesized association. Crude and adjusted odds ratios and 95% confidence intervals were estimated using conditional logistic regressions. Results suggest a protective role for serum ferritin for those in the highest quartile relative to those in the lowest quartile. Controlling for smoking and monthly personal income, an adjusted odds ratio of 0.2 with a corresponding 95% confidence interval of 0.1-0.7 was observed. Similar findings were noted when all other available CIN risk factors were controlled. In addition, a dose gradient was present for dietary iron intake (p = 0.01). No associations were observed between each of the other undernutrition indexes and CIN. Although only high levels of serum ferritin were associated with a protective effect against CIN, when coupled with the results from other studies that suggest carotenoids, folates, and vitamin C to be protective, the overall hypothesis that poor nutriture is associated with CIN remains viable. Lack of an association with the other nutritional indexes may reflect the relatively sufficient nutritional status of low-income individuals residing in the United States, as opposed to the undernourished population of the Third World. PMID- 8415131 TI - Oral and pharyngeal cancer, diet, smoking, alcohol, and serum vitamin A and beta carotene levels: a case-control study in men. AB - A case-control study was conducted in Melbourne, Australia. Forty-one men with histologically confirmed squamous cell oral or pharyngeal cancer were compared with 398 male community controls. A statistically significant increase in risk was found for alcohol (ethanol) consumption and for smoking, and there was a synergistic effect for these two exposures. Statistically significant protection was noted with increasing intake of dietary vitamin C, dietary beta-carotene, fruit, vegetables, and dietary fiber. The mean serum levels of beta-carotene and vitamin A were statistically significantly lower when the cases were compared with another set of 88 male controls of a similar age who were hospitalized for minor surgical operations. This study confirms a causal effect of smoking and alcohol and a protective role for a high dietary intake of fruit, vegetables, cereals, and, particularly, beta-carotene- and vitamin C-containing foods. PMID- 8415132 TI - Perceived change in food frequency among women at elevated risk of breast cancer. AB - Survey reports indicate that women perceive that their diets have changed in ways consistent with dietary guidelines recommended by national agencies. We have attempted to determine whether perceived change in food intake is a useful tool for estimation of either past intake or pattern of change in food consumption. Twin sisters of breast cancer patients, at obvious high risk of breast cancer, were aware of dietary guidelines concerning cancer, as indicated by beliefs about specific foods that should be increased or decreased to prevent cancer. Perception of change, as reported by them, was not found to be a reliable indicator of actual change in food frequency, as measured by the difference between sequential food frequency questionnaires. Consumption of high-fat foods was observed to have decreased over the interval, regardless of perceived change in consumption. Moreover, prediction of past food frequency on the basis of perceived change and current intake combined resulted in a less accurate appraisal of past diet than did the use of current intake alone. Perceived change in food frequency appears to be biased in different ways for different foods and seems to be influenced by beliefs about the role of diet in cancer. Studies of etiology should probably not rely on such methods. PMID- 8415133 TI - Ayurvedic agents produce differential effects on murine and human melanoma cells in vitro. AB - This study was performed to evaluate the relative efficacy of Maharishi Amrit Kalash ambrosia (MAK-5) and Maharishi Amrit Kalash nectar (MAK-4) on murine (B 16) and human (SK-Mel) melanoma cells in culture. Ethanol extract (EE) of MAK-5 (EE-MAK-5) induced morphological differentiation (enlargement of soma and nuclei and formation of long dendritic processes) and growth inhibition in mouse melanoma cells, whereas EE-MAK-5 inhibited only growth in human melanoma cells. Murine melanoma cells were more sensitive (about 3 times) than human melanoma cells in culture to EE-MAK-5; the aqueous extract (AE) of MAK-5 (AE-MAK-5) was ineffective in both cells. Boiling EE-MAK-5 for 10 minutes or exposing it to light at room temperature for 72 hours did not alter growth-inhibiting potency. Ethanol extract of another herbal agent, MAK-4 (EE-MAK-4), inhibited growth in human melanoma cells but not in mouse melanoma cells. AE-MAK-4 was ineffective for both cells. These results suggest that murine and human melanoma cells respond differently to MAK-5 and MAK-4 and that human melanoma growth-inhibiting agents are present in both EE-MAK-5 and EE-MAK-4. PMID- 8415134 TI - Effects of high-fiber diets on pathological changes in DMH-induced rat colon cancer. AB - The protective role of dietary fibers on tumorigenic effects of 1,2 dimethylhydrazine (DMH) on the rat colon was studied using histochemical, immunohistochemical, and biochemical analyses. Rats were injected with DMH (20 mg/kg sc) for five weeks, once a week, and were fed laboratory chow (Control I), semisynthetic fiber-free diet (Control II), or 18% or 25% assorted fiber (cellulose, beans, corn) diet. The rats were sacrificed 12, 16, and 24 weeks after the carcinogenic injections. Adenomatous tumors developed in 90-100% of the animals fed chow or fiber-free diets. A diet with high concentrations (25%) of corn dietary fiber significantly decreased the tumor incidence (40% and 42%) and tumor yield (p < 0.01). Diets with lower concentrations of fibers (18%) did not protect against tumorigenic effects of DMH. No differences were found in pathological parameters of tumors obtained from different dietary groups: the damage index (percentage of damaged cells per section) was very similar in all groups studied. Diets enriched with dietary fibers (25%) had a significant protective effect in DMH-induced rat colon cancer. PMID- 8415135 TI - Care and cure as healing processes in nursing. AB - Today's health care requires more than technology to accomplish healing. Nursing has a heritage of healing that was highly valued in the past. The care-versus cure controversy during the past two decades has not identified a clear healing role in contemporary nursing practice. The author explores the concepts of care, cure, and healing as interdependent and indivisible processes within health care and particularly within nursing. PMID- 8415136 TI - Toward a science of nursing--a method for developing unique content in nursing. 1964. PMID- 8415137 TI - Settling an ongoing debate with one entry level. AB - It is time that nursing settle this controversy regarding entry level to practice. In order to obtain full colleagueship with other professionals, improve the quality of care and the image of nursing, and receive a sound educational base, nursing should require the Bachelor of Science in Nursing degree as the minimal entry level for practice. PMID- 8415138 TI - Are nurses sexists? PMID- 8415139 TI - Use of the code of ethics for accountability in discharge planning. AB - Nine selected items from the Code of Ethics for nurses are used to illustrate how the code can be used as an excellent resource for accountability with discharge planning. Research findings identify the need for expanded awareness of the code and how to apply it to the discharge planning process. Components of the teaching/learning process are compared to the code to suggest that nurses need increased educational preparation to be accountable for discharge planning. PMID- 8415140 TI - Nursing education for better health: the vision beyond care. PMID- 8415141 TI - Protecting, promoting and supporting breast feeding: the special role of maternity services. PMID- 8415142 TI - Administration of nursing services in Rajasthan State: a report. PMID- 8415143 TI - An economy of words. PMID- 8415144 TI - Reader comments on article. PMID- 8415145 TI - Reader suggests family's role should be considered in PAD. PMID- 8415146 TI - Religiousness and hope in Hispanic- and Anglo-American women with breast cancer. AB - Religiousness has been associated with coping with cancer in the general population, but cultural influences have not been well explicated. The purpose of this study was to compare a sample of Hispanic-American women to a matched sample of Anglo-American women on selected religious variables and on a measure of hope. A sample of 25 Hispanic and 25 Anglo women diagnosed with breast cancer completed a hope scale, a spiritual well-being scale, and a religiousness scale and responded to selected demographic and medical questions. The only significant difference between the two groups was in intrinsic religiousness, with Hispanic women scoring higher (t = 2.07, df = 24, p < 0.05). Among Hispanics, neither intrinsic nor extrinsic religiousness was more important in predicting either existential well-being or hope. However, intrinsic religiousness was a more important predictor of religious well-being and total spiritual well-being than was extrinsic religiousness. Among Anglos, intrinsic religiousness was a stronger predictor of spiritual well-being and of hope. Religiousness may be an important variable affecting both the spiritual and the psychological health of women with breast cancer; this study also suggests cultural differences. PMID- 8415147 TI - The influence of guided imagery on chemotherapy-related nausea and vomiting. AB - The purpose of this study was to determine if the addition of guided imagery to a standard antiemetic regimen decreased nausea, vomiting, and retching occurrence and distress in patients receiving cisplatin-based chemotherapy. A convenience sample of patients (N = 28) was selected from an oncologist's patient population and randomized into two groups. Both groups received the same standard antiemetic regimen, while the experimental group additionally used a chemotherapy-specific guided-imagery audiotape. The Rhodes Index of Nausea and Vomiting Form 2 was used to measure the nausea and vomiting experience. Findings revealed no statistically significant difference in this measurement between the two groups when measured at five different times during chemotherapy administration. The Chemotherapy Experience Survey was used to evaluate the participants' overall perceptions of the chemotherapy experience. The guided-imagery group expressed a significantly more positive experience (p = 0.0001) with chemotherapy. These findings have definite implications for developing effective nursing interventions to promote patient involvement in self-care practices and to increase patient coping abilities during symptom occurrence. PMID- 8415148 TI - Freedom of choice: an issue concerning alternate therapies chosen by patients with cancer. AB - Patients with cancer who choose alternate therapies often use limited sources of information from lay literature to influence their therapy decisions. The biomedical assessment of these alternate therapies usually is not made available to patients in the lay literature; therefore, patients' true freedom of choice must be questioned. This paper discusses alternate therapies in light of two recent ethnographic studies that examined the alternate-care beliefs and practices chosen by hospital patients in Canadian prairie cities. The alternate cancer therapies that patients have chosen are presented along with the biomedical perspective of these therapies. The patients' own descriptions of their searches for cures are included, and implications for nursing are provided. PMID- 8415149 TI - Functional status and hope in elderly people with and without cancer. AB - The relationships and differences in hope and functional status in elderly people with and without cancer were studied to enhance knowledge concerning these patients' quality of life. A conceptual framework was established using Roy's Adaptation Model of nursing and hope was measured as an indicator of adaptation using Miller's Hope Scale (MHS). In addition, functional status was assessed by six domains using the Philadelphia Geriatric Center's Multilevel Assessment Instrument (MAI). The with-cancer group was a convenience sample of 86 patients 65 years of age and older with cancer who were attending an oncology clinic. The without-cancer group consisted of 88 elderly people in the community who perceived their health as good or excellent and who had never been diagnosed with cancer. Using a regression model, physical health was the only MAI domain, along with the demographic variables of income and education, that was related significantly to MHS scores. The results of this study indicate that declining physical health is a threat to hope and that lower socioeconomic status may be a threat to hope; however, age, gender, or a diagnosis of cancer are not. PMID- 8415150 TI - Central venous catheter "pinch-off" and fracture: a review of two under recognized complications. AB - Although uncommon, "pinch-off syndrome" and catheter fracture are reported complications of central venous catheters (CVCs). Pinch-off syndrome is characterized by intermittent catheter malfunction in conjunction with radiologic evidence of catheter compression. Warning signs of pinch-off syndrome include difficulty with-drawing blood samples and resistance to infusion of IV fluids. CVC fracture is characterized by migration of the distal catheter fragment through the heart and, often, into the pulmonary artery; it may be accompanied by the sudden onset of chest pain, palpitations, and arrhythmias. Twenty-seven cases of CVC fracture were reviewed, including 22 cases reported in the literature as well as an analysis of 5 cases that occurred at the authors' institution. Among the 22 cases reported in the literature, the average length of time between catheter insertion and fracture was 6.7 months. In 82% of these cases, the fracture occurred at the clavicle/first rib junction, where mechanical friction against the catheter has been well established as the mechanism for most fractures. In 9 of these 22 cases, evidence of catheter compression was noted on chest x-ray prior to fracture. This paper discusses assessment criteria for pinch off syndrome and catheter fracture, as well as nursing implications regarding the prevention and early detection of these potentially serious complications. PMID- 8415151 TI - Influences of nutrition and stress on people at risk for neutropenia: nursing implications. AB - Neutropenia may be influenced by malignancy type, treatment, age extremes, inadequate nutrition, or psychological stress. Of these five factors, only nutrition and stress are amenable to nursing intervention and management. The increasing trend of providing treatment in the outpatient setting and managing the patient with neutropenia in the home challenges nurses to develop innovative methods of care. This article offers suggestions to assist nurses in the creative management of individuals at risk for neutropenia by maximizing nutrition and minimizing psychological stress. This discussion addresses the physiology of the inflammatory immune response; pathophysiology of neutropenia; factors that may influence the risk of infection, such as sustained stress, dietary fiber, antioxidant vitamins, and food-borne bacteria; and interventions that reduce the potential for neutropenic sepsis. Nursing implications that reduce the risk of neutropenic infection include patient education related to nutrition, stress management, and self-care. PMID- 8415152 TI - BSE Rap: intergenerational ties to save lives. AB - This article presents an innovative public-education strategy that was created to promote breast health awareness and early breast cancer detection among minority and low-income adolescent females. Given the importance of teaching breast self examination (BSE), program development focused on creation of the BSE Rap, a lively music-video presentation. Increasing adolescents' knowledge and awareness of BSE is viewed as a springboard for disseminating information to their mothers and grandmothers. Funding was obtained for production of a video and a breast health diary, which are the program's key components. Marketing strategies included contacts with community organizations and healthcare professionals. Program evaluations reveal that the BSE Rap serves as a positive motivator for participants to discuss BSE and mammography with their mothers and grandmothers. The BSE Rap offers oncology nurses the opportunity to save lives using a unique and creative tool that focuses on intergenerational ties. PMID- 8415153 TI - Community screening programs made more effective. PMID- 8415154 TI - Metal increases radiation therapy-related mucositis. PMID- 8415155 TI - New oral care products treat radiation therapy discomfort. PMID- 8415156 TI - Antifungal agent used topically to control odor. PMID- 8415158 TI - Cancer resources in the United States. PMID- 8415157 TI - Yogurt helps to control wound odor. PMID- 8415159 TI - ONS Chapter Directory. PMID- 8415160 TI - ONS Special Interest Group (SIG) Directory. PMID- 8415161 TI - Microhardness and porosity of Class 2 light-cured composite restorations cured with a transparent cone attached to the light-curing wand. AB - A new technique for curing class 2 composite fillings was investigated with respect to microhardness and porosity in the cervical part of the restorations. The technique is based on a plastic transparent cone that is attached to the curing wand. Before polymerizing the cervical portion, the cone is pressed down into the material in the direction of the floor of the approximal box. When the light is activated, it concentrates in the tip of the cone, from where it is distributed into the composite material. In order to compare this technique with conventional curing, standardized class 2 cavity preparations were made in brass blocks. Four different composite materials were used for 20 restorations each: Heliomolar, Herculite XR, Occlusin, and P-50. Ten restorations of each material were cured using the conventional technique (in two portions), and in the remaining 10 restorations the cervical portion was cured with the transparent cone. The irradiation time was 60 seconds in all instances. The Vickers hardness of the cervical approximal surfaces was measured after one and 24 hours. The surfaces were photographed in a stereomicroscope, and the numbers of porosities were counted on black-and-white prints.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415162 TI - Use of a caries-disclosing agent to improve dental residents' ability to detect caries. AB - Clinicians often have difficulty determining which dentin should be removed during cavity preparation. Caries-disclosing agents have proven to be useful in the identification of carious dentin that requires removal. Recent dental school graduates are able to improve caries identification and removal when a caries disclosing dye is used to check their attempts at complete caries removal. PMID- 8415163 TI - Improving casting contours by regaining horizontal space. PMID- 8415164 TI - Clinician of the Year Award. PMID- 8415165 TI - Distinguished Member Award. PMID- 8415166 TI - Award of Excellence. PMID- 8415167 TI - Reasons for placement, replacement, and age of gold restorations in selected practices. AB - A total of 745 gold restorations were surveyed concerning the reasons for their placement and replacement. Cast gold restorations (n = 480) were placed due to primary caries (23%), replacement of amalgam (39%), and composite restorations (9%), or due to failure of existing gold restorations (29%). The main reasons for replacement of cast gold restorations were fracture of tooth (36%) and secondary caries (22%). The main reasons for the placement of compacted gold restorations (n = 265) were treatment of primary caries (40%), failure of an existing compacted gold restoration (26%), removal of another type of restoration (18%), and repair of a defective margin of a casting (16%). The main reasons for replacement of compacted gold restorations were inclusion into larger restorations (29%) and secondary caries (21%). The longevity of failed gold restorations was recorded for 111 restorations. The mean age for failed cast and compacted gold restorations was about 18.5 years (range 5-41 years). The ages of 2564 gold restorations in situ, 1689 castings, and 875 compacted gold restorations were recorded. The median and mean ages for gold castings were about 15 and 16 years and for compacted gold restorations about 17 and 18 years. PMID- 8415168 TI - Retention of Class 3 composite restorations: retention grooves versus enamel bonding. AB - This study compared the tensile loads required to dislodge class 3 composite restorations with and without retention grooves. Thirty extracted human maxillary central incisors were divided into two groups of 15 teeth each. A C-shaped class 3 preparation with lingual access was cut into one approximal surface of each incisor to standardized dimensions using a #329 bur. A 0.5-1.0 mm 45 degrees bevel was prepared on the lingual and gingival enamel margins. In one group, an incisal retention point and a gingival retention groove were prepared with a #1/4 round bur to a depth of 0.25-0.5 mm; the other group had no retention grooves/points. A nonretentive 2 mm round "well" with diverging walls was cut 0.5 1.0 mm into the axial dentin to accommodate the head of a pin that was inserted prior to composite restoration. Each pin shaft extended approximally from its incisor. A bonding agent (Universal Bond 3 Primer and Adhesive, L D Caulk) was applied to each preparation, and composite resin (Prisma APH, L D Caulk) was inserted incrementally. Each increment was exposed to 40 seconds of polymerization light. Restoration surfaces were finished and polished with Sof Lex disks (3M Dental Products). Specimens were thermocycled 6 to 60 degrees C, for 500 cycles, with a 30-second dwell time. They were then positioned in an aligning device, and pins were loaded in tension in an Instron Testing Machine at a head speed of 2 mm/minute to restoration failure. Mean (SD) failure loads in Newtons were: no grooves 83.6 (19.8); grooves 69.6 (18.1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415169 TI - A clinical comparison of glass ionomer (polyalkenoate) and silver amalgam restorations in the treatment of Class 5 caries in xerostomic head and neck cancer patients. AB - Fifty-four pairs of restorations (one glass ionomer and the other amalgam) were placed in the mouths of 36 xerostomic head and neck cancer patients. Patients were either fluoride users or fluoride nonusers. In patients who used a daily application of a mildly acidic (pH 5.8) sodium fluoride gel, glass-ionomer cements failed and amalgam restorations did not (P < 0.0001). In patients who neglected to use their topical fluoride as directed, glass-ionomer cement restorations did not fail, but amalgam restorations did (P < 0.001). The mean time to restoration loss for both restorative materials was 8.5 months. In severely xerostomic patients these findings were exaggerated. PMID- 8415170 TI - [Neonatal cholestasis: the viewpoint of the pathologist]. AB - The authors make a review of paediatric-neonatal cholestasis, a troublesome field in clinical and hepatopathology. They focus the main topics and stress the differential aspects in extrahepatic biliary atresia, "neonatal hepatitis", cholestatic syndromes associated with paucity of interlobular biliary ducts and metabolic diseases. The "overlap" areas and differential diagnosis in these chapters are a challenge for the pathologist. The importance of a close collaboration with clinicians is stressed. PMID- 8415171 TI - [Neonatal cholestasis: the viewpoint of the surgeon]. PMID- 8415172 TI - [Neonatal cholestasis: the viewpoint of the pediatrician]. AB - The heterogeneous nature of the diseases causing neonatal cholestasis, and the wide range of overlap among the specific entity frequently create serious challenges in their diagnosis. Moreover, the spectrum of correlated nutritional abnormalities in children with chronic cholestasis dictates a complex approach to their management. Nevertheless, pediatricians should be aware of the importance to recognize rapidly a specific treatable metabolic, infectious or malformative disease, in order to institute an early, specific, and effective treatment. However, despite all the still unanswered questions, in the last few years a tremendous progress has been achieved in the development of instruments and clinical procedures for diagnosis and therapy. Moreover, since orthotopic liver transplantation has been accepted as the treatment of choice of end-stage disease, a great deal of interest has been focused on the modalities of intervention applicable during progression of the disease. In this regard, nutritional support plays an increasing important role, since improved growth of these infants has a direct impact on their survival and suitability as candidates for liver transplant surgery. In the present article, Authors make an up-to-date of the main problems correlated with the diagnosis and management of chronic cholestasis. PMID- 8415173 TI - [The newborn infant of the diabetic mother: the clinical findings in 431 subjects]. AB - 431 newborns of diabetic mothers (NDM) were studied between the years 1980 and 1990. They were divided into two groups: a first group of 227 patients born from mother with gestational diabetes mellitus; a second group of 204 patients born from mother with pregestational diabetes mellitus. The first and the second group were subdivided into two subgroups: the A, referring to the period 1980-85, and the B, referring to the period 1986-90. Subgroups B were characterized by a better metabolic control of pregnancies. Our protocol consisted of anamnestic study, clinical, metabolic, instrumental examinations and clinical follow-up for a period ranging from 1 to 10 years. The study of our data suggests that embryo feto-neonatal mortality doesn't show significantly difference between the first and the second group and between subgroups A and B. The percentage of preterm infants was significantly more elevated in the second group, the percentage of full-term newborns was significantly higher in the first group. The incidence of cesarean sections was significantly higher in subgroups B. Mean birthweight was more elevated in the first group and the incidence of macrosomia was higher in subgroups A. The incidence of asphyxia, hypotonia, seizures, transient cardiomegaly, transient hypoglycaemia and hypocalcemia, was higher in subgroups A, without significant difference between the first and the second group. Congenital anomalies were heterogeneous and there were no significant difference between the first and the second group. Only the incidence of congenital cardiopathies was significantly more elevated in the second group. At follow-up neurologic sequelae were mainly characterized by behavioural anomalies. PMID- 8415174 TI - [Neonatal weight: an analysis of the distribution]. AB - We examined birthweight distribution in relation to gestational age from 25 to 42 weeks in a series of 3.526 single newborns and in whom reliability of gestational age was rigorously controlled. To verify the distribution normality the Shapiro Wilk and the Kolmogorov tests have been applied. Birthweight data follow a gaussian distribution for each gestational age week. Therefore in the birthweight standards estimation the parametric method can safely be applied also when the series includes pathologic and preterm newborns. PMID- 8415175 TI - [Specific IgE in the cord blood of infants from allergic parents]. AB - We evaluated the cord blood level of specific IgE in newborn having first grade relatives affected by allergic disease. We aimed: a) to look for a significant threshold (if any) of sp. IgE predictive of atopy; b) to verify the clinical reliability of the CAP System by Pharmacia. 9 out of 60 patients with positive family history had sp. IgE > 0.35 KU/L; 7 out of these 9 children were symptomatic to a clinical follow-up at 18 months. Moreover 65% of children with positive history but no sp. IgE and 100% of patients with both a negative history and no sp. IgE were completely free of allergic symptoms in the same period. We conclude that a careful history and cord blood sp. IgE measurement might be of value in early diagnosis of atopy. PMID- 8415176 TI - [The clinical significance of fetal hydronephrosis of moderate extent]. AB - Twelve fetuses with renal pelvis dilatation < 15 mm have been followed up until a year after birth. Dilatation cleared up in 59% of the cases within the last weeks of pregnancy and in 33% in the first months of life, while was asymptomatically detected in one case only a year after birth. A grade I and II vesico-ureteral reflux was diagnosed in two babies. Scheduled ultrasonographic examinations together with routine biochemical tests only are suggested by the kidney anatomical-functional normal parameters. Possible recurrent infections may be prevented through urine culture. Voiding cystourethrography is performed only when a recurrent infection is present because a reflux may be detected. Lacking a definite prognostic parameter, pre- and postnatal ultrasonographic follow-up is necessary although the little renal pelvis dilatation is to be considered negligible. PMID- 8415177 TI - [Elevated atrial natriuretic peptide (ANP) levels and normotension in Bartter's syndrome in childhood]. AB - The authors describe a case of Bartter's syndrome, a variety of disease in the newborn, and they point out the most specific of the disease symptoms, that is the normal PA in presence of elevated plasma renin activity. So it was necessary to determine the ANP, the values of which, controlled for a period of 18 months, appeared above the normal ones. The authors conclude that this date would better explain the apparently contradictory date of the normotension. PMID- 8415178 TI - [A pathogenetic hypothesis based on the use of chlorpromazine of organic disorders probably due to microcirculatory changes]. AB - In the introduction it is noted that, in the physiopathology, specific pathogenetic elements are missing concerning irritative stimulation, turbid fat pathosis, digital hippocratism of chronic affections (for example, pulmonary affections), the most frequent onset of telarche and of the swelling of the areola of the breast on the left hemithorax in the premenstrual syndrome, fibrosis, cyrrosis, certain types of insipid diabetes, etc. In the opinion of the author, the use of chloropromazine, in doses that have proved to be harmless, has contributed to the clearing up of some questions concerning a few pathologies of internal organs: the liver, the spleen, the brain-and enable us to pose some hypotheses about the swelling of the liver, the origin of scleroses and cirrhoses and some splenic and encephalic swellings. The author suggests that the fundamental reason is to be sought in changes in the microcirculation which are linked to insufficient capillary and sinusoidal circulation. Two cases of insipid diabetes are mentioned which were treated with chloropromazine and for which an improvement in the trophism of the diencephalic cells was hypothesized, due to an improvement in the local circulation. A similar physiopathological microcirculatory behaviour is attributed to digital hippocratism, the P. Marie and Bamberger syndrome (similar to those determined by cyanotic congenital cardiopathies), both due to chronic suppurative processes, and the slightly more frequent onset of telarche on the left hemithorax. It is expected that other pathologies may be explained by a similar physiopathological mechanism, malignant tumor inclusive. PMID- 8415179 TI - [Chronic diarrhea due to Cryptosporidium: the efficacy of spiramycin treatment]. AB - Cryptosporidium has received increasing attention as a pathogen in normal as well as immunocompromised persons. We report a case of cryptosporidiosis occurred in a 18-months-old-girl who was treated with spiramycin for 8 days with complete resolution of diarrhea within the first days of therapy; follow-up stool examination was negative for the protozoan. The incidence of diarrhea due to Cryptosporidium seems to be higher than it was reported before: so fecal samples, when negative for other etiological agents, should be examined for Cryptosporidium oocysts. Spiramycin seems to be an effective and well-tolerated drug: treatment is recommended in children with prolonged and/or severe diarrhea when Cryptosporidium seems to be the etiological cause of the disease. PMID- 8415180 TI - [Rheumatic manifestations and dental foci: a review of the cases in 12 years of activity in a pediatrics division]. AB - By examining 8244 clinical records, in a period of 12 years of paediatric activity, the authors point out the connection between dental caries and rheumatic fever. They suggest fluoride supplementation since the early age, in countries where the fluoride is lack in drink able-water. PMID- 8415181 TI - [Acute fetofetal transfusion in dichorionic twins: a clinical case report]. AB - Fetofetal transfusion is due to the twin to twin blood shunt caused by vascular anastomoses. It has been suggested that two types of FFT exist: a chronic form existing during pregnancy, and an acute form occurring only during parturition. Cross-circulation may be demonstrated in monochorionic as well as dichorionic placentas. However, such communications were found in 85-100% of monochorionic placentas, compared with one of 68 dichorionic fused placentas. Authors describe one case of acute feto-fetal transfusion in dichorionic twins. PMID- 8415182 TI - [Pulmonary arteriovenous fistula: a case in childhood]. AB - The authors describe a rare case of congenital, bilateral arteriovenous fistulas of the lung, without other symptoms of hemorrhagic telangiectasis. The disease was diagnosed in a ten-year-old child, because of a chronic hypossiemia state. The angiography with digitalized technique defined, at the best, the vascular anomalies; the examen cannot be substituted by other imaging methods. First a therapeutic embolisation in angiography was tried without success because of the dimension of the arteriovenous shunt. Then a local excisional therapy was carried out, on one lung. One year after the operation, the oxygenative and clinical conditions of the patient are excellent. PMID- 8415183 TI - [An anomalous vena cava inferior with azygos continuation: a clinical case report]. AB - We report a case of azygos continuation of inferior vena cava. The diagnosis is confirmed by C.T. scans. We also report possible diagnosis among other pathologies which substain similar radiographic pictures bat which are connected with silent clinic evolution. PMID- 8415184 TI - [Traumatic extradural hematoma in a newborn: a case report]. AB - The authors report the case of a twenty-six-day-old patient presenting a traumatic extradural haematoma. The main clinical aspects of this feature are illustrated and the approach to diagnosis and management is discussed. PMID- 8415185 TI - [Axial polydactyly: a clinical case]. AB - After a short review of the normal embryonal development of lower limbs the authors describe a case of polydactyly associated with a 2nd-grade hypospadias observed in a 4-month-age patient. The authors, after the surgical treatment of both defects, have made the follow-up of the patient until six year age. PMID- 8415186 TI - [Current advances in the cytogenetics and molecular biology of the diagnosis of malformation syndromes]. AB - Consistent progress has been obtained in recent years in the diagnosis and understanding of genetic disease. High resolution chromosome analysis has narrowed the gap between cytogenetics and Mendelism by proving that some diseases considered to be due to monogenic inheritance result in fact from deletion of contiguous genes or breakage of a specific gene. Gene "imprinting" has been found to be the cause of some syndromes, such as the Prader-Willi, Angelman, and Beckwith-Wiedemann. Molecular cytogenetics has become a tool for analysing small rearrangements not easily seen using traditional chromosome analysis, for detecting mosaicisms, and narrowing the "critical" regions of chromosomal syndromes. "Positional cloning" techniques have improved our ability to map disease-genes. Linkage studies and direct analysis of cloned genes are now having a large impact on the analysis of genetic diseases segregating in families, and in testing for healthy heterozygotes and presymptomatic patients. Discovery of mutations in the mitochondrial genome has widened our understanding of disease inheritance. A few disorders have been shown to be due to tandem amplification of GC-rich intragenic triplets. Genetic counseling has taken advantage of these improvements which have a large impact on our knowledge of genetic heterogeneity and variability. PMID- 8415187 TI - [The importance of the clinical record in the follow-up of malformation syndromes]. AB - In view of the pediatrician's need to know the natural history of these patients so as to guarantee appropriate clinical and instrumental follow-up it is necessary to collect information, in a clinical file and in as much detail as possible, on the classic phenotype and the different clinical manifestations of the various syndromes. The aim of this is to identify the most suitable times for applying possible therapy, also bearing in mind the increase in the mean age of these patients. PMID- 8415188 TI - [The assessment of neuropsychic aspects in dysmorphic syndromes (an introduction to the discussion)]. AB - The author reports brief notes on the neuropsychic aspects of dysmorphic syndromes. He suggests a diagnostic and follow-up protocol for the evaluation of learning capabilities, linguistic development and personality. PMID- 8415189 TI - [The evolution of epilepsy in the most common genetic forms with mental retardation (Down's syndrome and the fragile X syndrome)]. AB - Some chromosomal abnormality syndromes carry a higher risk of seizures than that found in the general population. Down's syndrome is considered to be the first and the most frequent chromosomal abnormality causing mental retardation. In spite of numerous reports and epidemiological surveys, the outcome of epileptic syndromes in patients with Down's syndrome (DS) is still largely unknown. We retrospectively studied 34 DS patients with epilepsy (14M; 20F). Epileptic syndromes were classified as: infantile spasms, 10 cases, i.e. 31%; Lennox Gastaut syndrome, 5 cases, 15.5%; symptomatic generalized epilepsy, 1 case; idiopathic generalized epilepsy, 6 cases, 17.6%; partial symptomatic epilepsy, 10 cases, i.e. 31%. In 2 patients the epilepsy was unclassifiable. In all the patients the following evolutive particularities were noted: a) the infantile spasms to have a relatively mild prognosis, as 8/10 patients remained seizure free, 3 of whom without treatment; b) no patient experienced febrile convulsions prior to the onset of epilepsy; c) Lennox-Gastaut syndrome had a relatively late onset (mean age 10 years, range 8-11.5); d) 7 patients (20.6%) developed reflex seizure. The fragile X syndrome is considered to be the second most frequent chromosomal abnormality causing mental retardation. The prevalence of epilepsy varies from 9.1% to 45% in the different series. In order to evaluate the prevalence rate of epilepsy and the previously hypothesized association with a particular electroclinical picture, we retrospectively studied 90 fragile X syndrome patients (80M, 10F) aged 4 to 25 years (mean age 13y6m).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415190 TI - [Cognitive and psychological profiles in dysmorphic syndromes]. AB - The recognizable patterns of human malformations have recently received much attention, particularly because of the decline of other diseases. Patients with a congenital malformation syndrome come to the Child Neuropsychiatrist for various reasons, such as: mental retardation of variable degree, learning disabilities, speech delay or absence of speech, behaviour disorders, various neurological impairment. Parents, however, seem to be mainly concerned about the prognosis of cognitive and psychological aspects. We have studied 83 patients with a specific pattern of malformations (35 affected by the Sotos syndrome; 25 by the Williams syndrome; 9 by the Cohen syndrome; 8 by the Cornelia De Lange syndrome; 6 by the Rubinstein-Taybi syndrome) and have particularly investigated their cognitive and psychological profiles. 13/83 showed a normal cognitive level (9 Sotos syndrome; 4 Williams syndrome), while 70/83 showed a cognitive deficit ranging from mild moderate (56 cases) to moderate-severe (14 cases). Linguistic deficits are prominent in the Sotos, Cornelia De Lange, and Rubinstein-Taybi patients, while practo-gnosic deficits are frequent in the Williams and particularly in the Cohen syndrome patients. The personality structure is characterized by immaturity and anxiety in all but the Williams syndrome patients, where some peculiar neurotic traits may be observed. All patients showed good communicative abilities. PMID- 8415191 TI - [The analysis of nervous system functions in dysmorphic syndromes]. AB - The involvement of the C.N.S. in dysmorphic syndromes is very frequent; therefore a systematic analysis of the functions of the nervous system is important in the clinical definition of these syndromes. Besides the morphological aspects, studied by magnetic resonance imaging, investigations should be carried out in the neuroelectrophysiological and neuropsychological fields. For the former, the following examinations are proposed: EEG in wakefulness and sleep, multimodal evoked potentials (VEP, BAEP, SEP), cortical magnetic stimulation and P300 (P3). For the neuropsychological field, a general intelligence test appropriate to the mental age of the subject (the Wechsler, Terman-Merrill, or Brunet-Lezine scale) and, whenever possible, the following complementary tests: Raven's Progressive Matrices, Bender's and Santucci's graphic tests, go-no go, Goodenough draw-a person, reading and writing tests, Langeot's scale for development of the logical thinking, sorting test and verbal and spatial memory tests. In some cases, the behaviour should be defined, through Conner's scale for attention deficit hyperactivity disorders, the Autism Diagnostic Interview, the Adaptive Behaviour Scale and the Brief Psychiatric Rating Scale. PMID- 8415192 TI - [Cognitive and neuropsychological profiles in malformation syndromes: a comparison between Down's syndrome and Williams' syndrome within a follow-up]. AB - The present work compares the neuropsychological and cognitive profiles of 4 children with the Williams and Down syndrome. The Authors stress the importance of an accurate analysis of each child's developmental story. They monitor children with physical malformations associated to Mental Retardation or other psychopathological problems in order to have a better understanding of their cognitive strategies and true potential and of their social and emotional implications. This procedure allows immediate action to be taken as soon as physical, emotional or cognitive problems arise. It also enables parents to be fully aware of their children's problems and to be able to meet their different developmental needs. PMID- 8415193 TI - [The incidence and prevalence of malformation syndromes]. AB - Data on the real incidence and prevalence of malformation syndromes are very scarce. The authors suggest the creation of "ad hoc" registers to improve the knowledge of these syndromes. PMID- 8415194 TI - [Malformation syndromes: the diagnostic approach and follow-up in ocular pathology]. AB - Ocular defects are often found in association with many systemic disorders. Some are already present at birth, but in other cases they develop in early infancy or later in life. They are found associated with connective tissue disorders, neurological conditions, chromosomal abnormalities and dermatological disorders. Some of these syndromes have a genetic basis but often it is uncertain whether they are inherited and their cause is unknown. PMID- 8415195 TI - [The current role of neuroradiology in evaluating malformation syndromes]. AB - CT and MRI play complementary roles in the study of malformation syndromes in pediatric age: CT in particular is indispensable for adequately assessing the skeletal picture in subjects suffering from cranio-facial malformations, while MRI allows a morphological study and supplies informations on the maturation state of the white matter. PMID- 8415196 TI - [Dental pathology in malformation syndromes: the methodology and intervention timing]. AB - Due to the complexity and potential vulnerability of the cranio-facial area (skull, face, nose, oral cavity), specific alterations of this region are associated to genetic and acquired malformations in a high percentage of cases (75%). Often, the specificity and extent of the pathological symptoms occurring in the cranio-facial area are the dominant aspects of the malformation syndrome according to which it is classified. When multiple development anomalies in a newborn suggest the existence of a syndrome, the patient should be placed in the care of a pediatrician, who, together with a team of specialists, will follow the child and face the various problems related to the syndrome at the right time. The frequent marked involvement of the craniofacial area in malformation syndromes suggests the opportunity of a specialist methodology to standardize odontostomatological therapy. PMID- 8415197 TI - [Malformation syndromes and deafness]. AB - Deafness is one of the most common clinical findings in patients affected by malformation syndromes. It may be congenital, neurosensory or transmissive in nature, or acquired as the consequence of phlogistic processes due to craniofacial malformations. It is important to identify and classify the hypoacusis in order to start therapy as early as possible, using surgery or prosthesis according to the indications, to avoid the child suffering both verbal and intellectual impairment. PMID- 8415198 TI - [The correction with plastic surgery of congenital malformations: the times for intervention]. PMID- 8415199 TI - [The usefulness of the skeleton in the diagnosis of malformation]. AB - Most malformation syndromes (70%) show evidence of skeletal anomalies, mainly in the vertebral bodies and in the bony segments of the limbs. This explains the importance of studying the skeleton when faced with any multi-malformation syndrome, using techniques like CT, MRI, echography and densitometry as well as standard radiology. To increase the quantity of information available for diagnostic purposes it would be profitable to create a data bank containing all the possible radiological findings encountered in the numerous multi-malformation syndromes. PMID- 8415200 TI - [The pathogenesis of cerebral malformations]. AB - Although data on the true incidence of brain malformation pathologies do not exist in Italy, such pathology is nevertheless becoming a question of current debate thanks to new neuroradiological and contrastographic techniques. For some of these brain malformations the time when the agent causes the anomalies and the various causative agents have been well identified, whether materno-foetal in origin or multifactorial. Subjects with brain malformations must be followed-up clinically with instrumental tests. PMID- 8415201 TI - [The natural history of syndromes with a metabolic origin]. AB - Notable progress has been recently made in the field of metabolic diseases, particularly for the therapeutic approach, which may be assessed in terms of survival, reproductive capacity and social adaptation. Therapy is often applied at different levels, e.g. the physical appearance, metabolism or only using medical symptomatic therapy. Positive results are expected from the administration of the functional protein, and from a possible treatment directly at the level of the somatic genotype. PMID- 8415202 TI - [Syndromes from birth to adulthood]. AB - A follow-up program of Down's syndrome is reported. Divided into age groups, it outlines the importance of a periodic clinical checks involving many disciplines and covering the entire lifespan of the patient. To do that we need a specific knowledge of the natural history of the syndromes. PMID- 8415203 TI - [Syndromes in outpatient experience]. AB - We report our experience as clinical geneticists and pediatricians in the management of patients affected by multiple congenital malformations and genetic syndromes. Our aim is to demonstrate that etiological diagnosis, crucial for successful management and counselling, has to be considered the first step in the care of these patients and their families. Coordination of medical interventions, clinical follow-up and psychosocial support have to be included in every out patient program in this field. Diagnostic approach has to be carried out through a specific methodology which includes extensive clinical expertise in dealing with children affected by multiple congenital anomalies, protocols for major clinical problems, facilities for specific investigations (cytogenetic laboratory, molecular genetics, ecc.), computerized programs for recognising rare syndromes, network of motivated specialists (neurology, ophthalmology, orthopedics, etc.) and possibility of collegial discussion of undiagnosed patients. Psychosocial support and follow-up have to be achieved through trained professionals. Our 5 years of experience in this field (1420 patients) suggest the need for an organized network of services. This could provide the families of children affected by genetic syndromes with a comprehensive management and support program which is lacking in the present health care system. PMID- 8415204 TI - [The therapy of malformation syndromes]. AB - The therapeutic possibilities in malformative syndromes are basically surgical, rehabilitative and, in a few cases, pharmacologic. The possibilities of using drugs are limited to some hormonal disorders caused by diencephalic and hypophyseal dysfunctions with clinical signs appearing at the level of the various target organs. The clinical signs most commonly found are hypogonadism, short stature and obesity. The Authors discuss on the opportunity of the use of the growth hormone in syndromic patients with short stature. PMID- 8415205 TI - [The follow-up of malformation syndromes: the new frontier]. AB - Much progress has been made over the last decade in the field of genetic diseases, mainly because of the advances in molecular biology. However, this has not been paralleled by similar progress in the assistance provided for such patients, owing to the lack of adequate follow-up of such chronic disorders. The author analyses the main objectives of the follow-up. PMID- 8415206 TI - [Syndrome X--associated diabetes]. PMID- 8415207 TI - [Behavior of C-peptide, insulin and glucose levels in blood during conditions of evaluating selected antihypertensive drugs in patients with hypertension and non insulin-dependent diabetes (type 2)]. AB - In 60 patients divided in three groups, each of 10 non-diabetic patients with essential hypertension (h) and of 10 hypertensive type 2 (non-insulin-dependent) diabetics (h+c), aged 31-63 years, the effect of 2-week treatment with nifedipine, captopril and prazosin on glycaemia, serum insulin (IRI) and C peptide (CP) after oral and i.v. glucose loading was compared. Nifedipine resulted in higher glycaemia levels in the oral test in both groups. This drug caused in group (h), but not in group (h+c), reduction of the glucose-dependent early increases of serum IRI and CP, more marked in respect to CP, what was expressed by the decrease of the serum CP:IRI ratio. These results prove that in non-diabetic patients nifedipine reduces the early response of the B-cells to glucose, but this effect is partly compensated by decreased insulin uptake by the liver. In patients with type 2 diabetes this phenomenon has not become manifest because of absence or reduction of early glucose-dependent insulin release. After captopril in both groups lower values of glycaemia and serum IRI and CP were found. Prazosin did not change the determined blood parameters. CONCLUSION: nifedipine, captopril, prazosin have a small influence on secretory function of pancreatic B-cells and may be recommended for the treatment of hypertension in patients with type 2 (non-insulin-dependent) diabetes. PMID- 8415208 TI - [Behavior of insulin, C-peptide and proinsulin levels in serum in the course of acute pancreatitis]. AB - 14 patients with acute pancreatitis during 16 episodes of the disease were studied. The concentration was measured of blood glucose, serum insulin (IRI), serum C-peptide (CP) using two methods: with Byk-Mallinckrodt kits and with more specific M-1221 antibodies Novo, and of serum proinsulin (Pro) in fasting state on days 1, 2, 3, 5 and 10 after the acute onset. Apart from some sporadic rises in the initial period of the disease, the blood glucose level did not change significantly, and had rather a decreasing trend. The mean serum IRI concentration was 0.17 +/- 0.17 (SD) nmol/l, and it decreased on the 5th day to 0.06 nmol/l 0.04 nmol/l, rising again to 0.11 +/- 0.15 nmol/l on the 10th day. The serum Pro concentration was on the same days: 11.1 +/- 12.6, 4.2 +/- 2.4 and 7.5 +/- 10.8 pmol/l, while the serum CP concentration determined with M-1221 antibodies was 0.48 +/- 0.50, 0.34 +/- 0.19 and 0.52 +/- 0.25 nmol/l respectively. However, when for serum CP determinations the Byk-Mallinckrodt kits were used, the concentration of this peptide was on the 1st day 1.90 +/- 1.12 nmol/l, and it decreased over the following days to 1.08 +/- 0.98 on the 5th day, but remained on the same level on the 10th day (1.11 +/- 0.46 nmol/l).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415209 TI - [Alcohol tolerance in patients with non-insulin-dependent diabetes (type 2) treated orally with drugs--derivatives of sulphonylurea]. AB - The oral ethanol loading test (0.5 g per kg b.m. given as 40% solution) was carried out in 5 groups, each of 10 patients with non-insulin-dependent (type 2) diabetes before and after 10 days of treatment with one of the following sulphonylurea derivatives: tolbutamide 0.5 t.i.d., chlorpropamide 0.5 once daily morning, glibornuride 0.025 t.i.d, glibenclamide 0.005 t.i.d. and glipizide 0.005 t.i.d. The response to alcohol (facial flush, heart rate, blood pressure) were compared, and blood concentration of ethanol, acetaldehyde, pyruvate, lactate, carbonates as well as blood pH, pO2 and pCO2 were determined in fasting state and during 6 hours after alcohol ingestion. In all patients the family history of diabetes and the presence and degree of vascular complications were registered. Evident flushing phenomenon was observed in 6 patients treated with chlorpropamide, in 3 treated with tolbutamide, in 2 treated with glibenclamide, in one receiving glibornuride and in none treated with glipizide. All drugs caused a greater rise of blood ethanol and acetaldehyde levels in relation to the control tests, but the difference reached statistical significance only in the group receiving chlorpropamide. Moreover, patients (pooled) with positive thermographic response had also significantly higher blood levels of ethanol and acetaldehyde during the second test. The ratio of acetaldehyde to ethanol concentration in blood (mumol:mmol) was not significantly changed in any group indicating parallel impairment of both steps of ethanol metabolism. All studied drugs intensified to a similar degree the alcohol-induced hypoglycaemia, but had no significant effect on the decrease of blood pyruvate level neither on the increase of blood lactate level. They didn't change the post-alcohol decrease of blood bicarbonate and pH, and didn't modify the behaviour of partial gas pressure. There was also no difference between pooled groups of patients with positive and negative thermographic reaction with respect to family history of diabetes and frequency and intensity of vascular complications. It is concluded that in patients with non-insulin-dependent (type 2) diabetes the second generation sulphonylurea derivatives are associated with lower risk of alcohol intolerance in case of its incidental ingestion in small amounts. The hypothesis of association of positive thermographic reaction to alcohol during treatment with sulphonylurea derivatives with more frequent occurrence of diabetes in family members and lower tendency to vascular complications was not confirmed. PMID- 8415210 TI - [Impairment of bronchial reactivity in patients with diabetes mellitus type I]. AB - Bronchial provocation tests (histamine and acetylcholine) were performed in 40 subjects (30 of them with type 1 diabetes, and 10 healthy volunteers) without any history of respiratory disease, not smoking and not taking any bronchodilating drugs. Bronchial reactivity was assessed using PC20 estimated spirographically by measuring FEV1. The patients were classified into three groups according to the duration of the disease: group I (0-7 yrs), group II (8-15), and group III (> 15). In all the three groups diabetes was at a similar degree of compensation, as evaluated by the mean circadian glycaemia, serum fructosamine and the Schlichtkrull Mw index. Bronchial reactivity to acetylcholine and histamine decreased with diabetes duration. Reaction to acetylcholine was statistically lower after 7 years of diabetes. An autonomic neuropathy was detected within the respiratory system, parallel to tachycardia at rest, alteration of the Valsalva test and orthostatic hypotension. PMID- 8415211 TI - [Immunogenicity of semisynthetic human insulin Novo--five year prospective studies]. AB - The aim of the study was to compare the immunogenicity of semisynthetic human and porcine monocomponent (MC) insulin Novo during--5-year observation. Thirty-one diabetic patients, never previously treated with insulin were randomly allocated to treatment with one of the two coded insulin preparations in a double blind trial. 15 patients aged 20-58 years (mean age--36 years) were treated with human insulin and 16 diabetics aged 19-61 years (mean age--34 years) with MC pork insulin. The insulin was injected twice a day (Actrapid plus Monotard). Serum insulin-binding antibodies were determined according to Christiansen (radioimmunoelectrophoretic method). The development of insulin-binding antibodies during the first year of observation was more rapid in patients treated with MC pork insulin but thereafter it was similar in the two studied groups. After 5 years of treatment with human insulin serum insulin-binding antibodies were found in 14 patients. The level of antibodies was very low (< 0.130 mU/ml) in 5 subjects, low (0.171-0.401 mU/ml) in 6 patients and relatively high (0.885; 1.186; 5, 162 mU/ml) in 3 patients. In the group of diabetics treated with MC pork insulin after 5 years of observation serum insulin-binding antibodies were found in 12 patients. The level of antibodies was low (0.131 0.576 mU/ml) in 9 subjects and relatively high (1.034; 3.954; 5.639 mU/ml) in 3 patients. The results obtained after 2-5 years of the study did not differ significantly (Wilcoxon's test, p > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415212 TI - [Personal experience with combined treatment of gliclazide and insulin in patients with non-insulin-dependent diabetes (type 2) and late secondary failure of sulphonylureas]. AB - The study was carried out on 20 patients, aged 20-60 years, with non-insulin dependent (type 2) diabetes of 3-17 years duration and with secondary failure of sulphonylurea derivatives. In the initial run-in-period (8 weeks) the patients were treated only with insulin in a dose decreasing glycaemia in fasting state to 160 mg% (8.9 mmol/l), then during 16 weeks they received insulin and gliclazide 160-320 mg per day, and finally during the final period (8 weeks) they were transferred again to exclusive insulin therapy. The association of gliclazide with insulin resulted in better metabolic control expressed by significant decrease of basal and postprandial glycaemia and small (non-significant) decrease of serum triglycerides level--with simultaneous reduction of daily insulin requirement by 34% on the average. In this period an increase of basal and stimulated (test meal) C-peptide secretion was observed. The improvement of the metabolic state and the increased C-peptide secretion persisted also after gliclazide withdrawal (during the second period of exclusive insulin therapy), although the determined parameters were less favourable than when combined treatment was applied. The mechanism of metabolic improvement of patients with type 2 diabetes and secondary failure of sulphonylurea derivatives, which follows the addition of gliclazide to insulin, is not fully clarified. Probably it is the consequence of both, the increased secretion of endogenous insulin and the enhanced activity of this hormone in insulin-sensitive tissues. PMID- 8415213 TI - [Amylin--new peptide from B-cells of pancreatic islets]. PMID- 8415214 TI - Hemodynamic correlates of clinical severity in isolated ventricular septal defect. AB - To test the hypothesis that the clinical assessment of severity in ventricular septal defect would be more related to variables which define tissue oxygen delivery than variables which define the left-to-right shunt, cardiac catheterization data from 40 children < 3 years of age were assessed. Variables which were considered indicative of clinical severity included the need for digoxin and diuretics, resting heart rate, and severity of growth failure. Variables measured at cardiac catheterization, including those which related to oxygen transport, and assessment of left-to-right shunt, were considered independently. Patients receiving digoxin and diuretics were more tachycardic (142 +/- 18 vs. 111 +/- 26 beats/min, p < 0.001) and had lower superior vena cava oxygen saturation (64 +/- 6 vs. 69 +/- 5%, p < 0.01). Variation in heart rate (r2 = 0.46) was best explained by oxygen consumption, hemoglobin concentration, cardiac index, and pulmonary vascular resistance. Variation in growth failure (r2 = 0.15) was related only to the left ventricular forward stroke index. These data suggest that variables related to oxygen delivery, including oxygen consumption, hemoglobin concentration, cardiac index, forward stroke index, and superior vena cava oxygen saturation, are the major contributors to the clinical assessment of severity in ventricular septal defect. PMID- 8415215 TI - Heart rate variability in diabetic children: sensitivity of the time- and frequency-domain methods. AB - Heart rate variability (HRV) is a noninvasive index of the neural activity of the heart. Although also influenced by the sympathetic activity of the heart, HRV is essentially determined by the vagal stimulation of the heart. Several HRV abnormalities have been described in adults with diabetes mellitus. However, there are few data on HRV in children with diabetes mellitus. In the present study, HRV was assessed in seven healthy children, 10 diabetic children with good glycemic control and 11 diabetic children with poor glycemic control. All had normal standard cardiac autonomic function tests, obtained from 24-h Holter tapes. HRV was measured by calculating six time-domain (mean R-R interval (RR), standard deviation of the R-R interval [SDRR], standard deviation of the mean of 288 R-R intervals [SDANN], the mean of the 288 standard deviations computed for each 5-min period [SD], percentage of differences of adjacent R-R intervals of > 50 msec for the entire 24 h [pNN50], and the root mean square of successive differences [rMSSD]) and four frequency-domain (low frequency [LF], high frequency [HF], total heart rate power spectra, and LF/HF ratio) indexes. SD, pNN50, rMSSD, LF, HF and total heart rate power spectra were markedly and significantly reduced in diabetic children with poor metabolic control. The 24-h variation of low- and high-frequency components of heart rate power spectra of the latter children had a different shape. Thus, diabetic children with poor metabolic control (elevated HbA1c and B2M levels) have a low HRV compared to those diabetic children with good control and healthy children.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415216 TI - Ambulatory arrhythmia screening in symptomatic children and young adults: comparative effectiveness of Holter and telephone event recordings. AB - Effective initial identification of potential cardiac arrhythmias in symptomatic children is difficult due to the infrequency, brief duration, and vague nature of subjective complaints in the young. Although both telephone event and Holter monitoring are used for this purpose, no comparative studies of the initial screening efficacy of either have been performed. A total of 202 consecutive symptomatic children (age 11 days to 26 years, mean 10.2 years) were evaluated for potential cardiac arrhythmias with either 24-h Holter (97 patients) or telephone event (105 patients) recorders and grouped according to the presence or absence of congenital heart defects, normal or abnormal resting electrocardiogram (ECG), and presence or absence of cardiac surgery. The results showed 30% of all recordings (61% event; 14% Holter) failed to substantiate any arrhythmias in spite of subjective symptoms. Event recordings showed a better correlation of sensed symptoms with arrhythmias (32%) compared to Holters (5%) (p < 0.01) with 73% of Holter recordings performed during both asymptomatic and arrhythmia-free 24-h periods. Holter monitoring was more effective in detecting nonsensed and asymptomatic events (8% versus 0.5%, (p < 0.01), among high-risk children. This study demonstrates that although both monitoring devices are applicable to children, each has inherent limitations and usefulness. These must be considered in choosing either device to permit their most optimal and cost-effective application. PMID- 8415217 TI - Effect of patent ductus arteriosus on Doppler-derived right ventricular systolic time intervals. AB - Right ventricular systolic time intervals (RVSTI) and noninvasive Doppler-derived pulmonary blood flow were measured before and after surgical ductus ligation in 18 otherwise healthy infants and children who were older than 3 months of age. Right ventricular preejection period (PEP) and the ratio of preejection period and right ventricular ejection time (PEP/RVET), both corrected or uncorrected for heart rate, decreased significantly following surgery (PEP 71 +/- 14 vs. 50 +/- 13, p < 0.001 and PEP/RVET 0.29 +/- 0.06 vs. 0.21 +/- 0.05, p < 0.001). The volume of pulmonary blood flow correlated with PEP/RVET (r = 0.48, p = 0.003). The magnitude of the change in pulmonary blood flow correlated with the change in PEP/RVET (r divided by 0.56, p = 0.016). The velocity of circumferential fiber shortening (VCFc) increased after surgery, but not significantly. We speculate that patent ductus arteriosus has a similar effect on right ventricular performance when other congenital heart defects are present. PMID- 8415218 TI - Aberrant right subclavian artery with left aortic arch: associated cardiac anomalies. AB - Anomalous origin of the right subclavian artery (ARSA) from the aorta distal to the normally positioned left subclavian artery is a relatively frequent congenital anomaly in subjects with left aortic arch. The purpose of this study was to determine the relative frequency of associated cardiovascular anomalies in individuals with this anomaly. From the records of approximately 11,000 pathologic specimens in the Registry of Cardiovascular Disease of United Hospital (St. Paul, MN, USA), we found 128 (1.2%) with ARSA. Of the 128 ARSA, 117 (2.9%) occurred among 4102 instances of congenital heart disease. The 117 cases with congenital heart disease and ARSA were conotruncal anomalies in 38%, septal defects in 28%, obstructive anomalies of the left side of the heart in 21%, right heart anomalies in 5%, and miscellaneous conditions in the other 8%. Down syndrome existed in 14 (12%) of the 117 specimens with ARSA and some congenital cardiac anomaly; nine of the latter had an atrioventricular canal (AVC) malformation. PMID- 8415219 TI - Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) decrease in diastolic left ventricular function assessed by echocardiography. AB - Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) are known to be associated with cardiomyopathy. Systolic and diastolic left ventricular functions were assessed by M-mode and Doppler echocardiography in four patients with MELAS and in 14 normal controls. The interventricular septal thickness and left ventricular posterior wall thickness were greater (11.0 +/- 1.6 mm vs. 5.8 +/- 0.7 mm and 11.0 +/- 2.2 mm vs. 5.9 +/- 0.8 mm) in patients with MELAS than in a control group. Parameters of systolic left ventricular functions (ejection fraction, shortening fraction, systolic time intervals, and mean Vcf) and left ventricular dimensions were not significantly different between the two groups. To assess the diastolic function, blood flow velocity across the mitral valve was measured by Doppler echocardiography and various indexes were obtained. In patients with MELAS, the impairment of diastolic left ventricular filling was demonstrated by decrease in the following indexes: peak flow velocity in the early passive filling period (E) (0.76 +/- 0.10 m/s vs. 0.94 +/- 0.09 m/s), integrated velocity for total E (10.2 +/- 1.3 vs. 13.0 +/- 0.9), the ratio of E and late atrial filling integrated velocities (1.72 +/- 0.06 vs. 2.49 +/- 0.29). PMID- 8415220 TI - Echocardiographically guided balloon atrial septostomy during extracorporeal membrane oxygenation (ECMO). AB - Use of extracorporeal membrane oxygenation (ECMO) in infants with congenital heart disease is becoming more frequent. We present the first reported use of balloon atrial septostomy during ECMO support and describe possible complications of such procedures unique to ECMO therapy. PMID- 8415221 TI - Coronary arterial lesions due to Kawasaki disease: selective coronary angiography in five cases with difficult-to-detect localized stenosis. AB - We report five cases of localized stenosis in coronary arterial lesions due to Kawasaki disease which were difficult to demonstrate by routine selective coronary angiography. Of these cases, three had localized stenosis overlapping the aneurysm, one had localized stenosis overlapping another adjacent branch, and the fifth had localized stenosis at the proximal left main trunk of the coronary artery. The reasons for difficulty in demonstrating the stenoses by routine study include: the localized stenosis was often superimposed on the aneurysm and/or the other adjacent branches, and the catheter was pushed into the inlet of the aneurysm on the proximal left main trunk. For a precise demonstration of a localized stenosis by selective coronary angiography, many angiograms from different perspectives should be taken. In addition, when a large aneurysm exists in the left main coronary artery, selective coronary angiography should be taken without pushing the catheter into the inlet of the aneurysm, and each frame of the cine coronary angiography should be carefully examined. Care should also be taken to compare with the initial view of the projection. PMID- 8415222 TI - Congenital coronary artery fistula suggesting a "steal phenomenon" in a neonate. AB - The case of a neonate reported herein demonstrates a congenital fistula between the left circumflex coronary artery and the right ventricle. Intraoperative stress to the patient's heart suggested a coronary "steal phenomenon." PMID- 8415223 TI - Transcatheter closure of congenital coronary arterial fistula with a detachable balloon. AB - Coronary arterial fistulae are rare congenital cardiac defects that typically are treated by surgery. A case of transcatheter closure of a left anterior descending coronary artery to right ventricular fistula with a detachable balloon is described in a 16-month-old child. The fistula was easily occluded without complication. Follow-up 1.5 years later revealed normal ventricular function and no recurrence of the fistula. Detachable balloon occlusion of coronary arterial fistula is feasible in patients as young as 4 months. PMID- 8415224 TI - Coronary artery fistula complicating the evaluation of Kawasaki disease. AB - Two patients clinically diagnosed with Kawasaki disease were found to have a coronary artery to pulmonary artery fistula. The dilemma of deciding the etiology of coronary artery dilatation in these patients is discussed along with management. PMID- 8415225 TI - Primary hypertrophic osteoarthropathy: another heritable disorder associated with patent ductus arteriosus. AB - Patent ductus arteriosus (PDA) is a congenital malformation that has been linked to diverse heritable and chromosomal disorders. Primary hypertrophic osteoarthropathy (HOA) is a rare heritable syndrome in which digital clubbing and periostosis become evident without any underlying illness. The objective of this article is to describe four patients with coexisting PDA and primary HOA. Surgical closure of the ductus showed no effect on the skeletal changes. Primary HOA should be included among the heritable disorders that may be associated with PDA. PMID- 8415226 TI - Endocarditis in neonatal intensive care unit. AB - The clinical spectrum of infective endocarditis (IE) in infants is examined in four infants between 3 and 9 months of age. None of the patients had signs of IE; all four had an anatomically normal heart. Echocardiograms showed echo-dense vegetations in the left side of heart in three cases and in the right side in one. Three of the four patients recovered after the episode of endocarditis. Three of the four patients had necrotizing enterocolitis in the neonatal period. The important predisposing factor was the presence of indwelling central catheter for intravenous nutrition. Unlike previously reported cases, coagulase-negative Staphylococci and Enterococci were important causative organisms in this high risk nursery population. PMID- 8415227 TI - Left ventricular pseudoaneurysm diagnosed by magnetic resonance imaging in a nine year-old boy. AB - An asymptomatic 9-year-old boy was found to have a cardiac murmur in a heart disease survey. Two-dimensional echocardiogram identified a large left ventricular pseudoaneurysm. Color flow imaging showed a transtunnel "to-and-fro" mosaic pattern between the pseudoaneurysm and left ventricle. ECG-gated magnetic resonance imaging demonstrated the exact size and related location of the left ventricular pseudoaneurysm and its ostium. Without cardiac catheterization or angiography, he was successfully operated on by closure of the communicating tunnel. PMID- 8415228 TI - Juxtaductal aortic atresia masquerading as coarctation. AB - Two cases of juxtaductal aortic atresia diagnosed as coarctation on clinical and Doppler echocardiographic grounds are presented. The misleading nature of the Doppler flow velocity characteristics in this condition is discussed and raises questions as to the source of these flow velocities in coarctation. PMID- 8415229 TI - Discrete subvalvular aortic stenosis in the Beckwith-Wiedemann syndrome. AB - Various congenital cardiac malformations have been described in patients with Beckwith-Wiedemann (BW) syndrome, including reversible obstructive subaortic stenosis in one patient. We herein present a case of a 2.5-year-old black boy with BW syndrome and discrete subvalvular aortic stenosis of the membraneous type. Such association of these two entities has previously not been documented. PMID- 8415230 TI - Diagnostic assessment. AB - Precision in diagnosis is the basis for treatment. The use of structured assessments of diagnosis, severity of illness, and side effects through various rating instruments likely will become of greater importance to clinicians in the next decade. PMID- 8415231 TI - Treatment of acute mania. AB - Treatment of acute mania requires attention to both specific and nonspecific antimanic medications. The choice of specific agents now includes lithium, carbamazepine, and valproate; and nonspecific agents include benzodiazepines, calcium channel blockers, alpha adrenergic agonists, as well as neuroleptic drugs. Treatment-resistant manic states are best treated by careful sequential strategies that may include polypharmacy and electroconvulsive therapy. PMID- 8415232 TI - Treatment of acute depression. AB - This article presents information needed by the clinician to diagnose and treat acute depression. The emphasis is on the pharmacologic treatment of depression. Both clinical and preclinical pharmacology of antidepressants are reviewed. The data presented will help the clinician make a rational choice of antidepressant for his or her patient. In particular, these data will help the clinician avoid or minimize certain adverse effects and drug-drug interactions. Guidelines for treating depression as well as strategies for treating the refractory patient are presented. PMID- 8415233 TI - Atypical depression. A valid clinical entity? AB - The history of atypical depression is summarized, and the results of several treatment outcome studies are reviewed. A number of clinical course, family, and biologic variables in patients with atypical depression are investigated, and these patients are compared with patients with other depressive conditions. The Atypical Depression Diagnostic Scale Question Book also is presented. PMID- 8415234 TI - Electroconvulsive therapy. AB - Modern ECT practice has evolved far from its beginnings more than 50 years ago. ECT is effective, safe, and rewarding in the clinical setting. This discussion complements the 1990 APA Task Force report and elaborates on some of the clinical and scientific factors that could not be fully addressed by the report. The future of ECT lies in understanding the mechanisms by which it relieves depressive symptoms and causes amnesia and related cognitive deficits. PMID- 8415235 TI - Continuation and maintenance pharmacotherapy for unipolar and bipolar mood disorders. AB - There is evidence that the risk for recurrence increases with each additional mood episode. Substantial data show that the probability of relapse and recurrence in both unipolar and bipolar disorders can be decreased by extended pharmacologic treatment. In recommending prolonged treatment to prevent recurrence, however, the values of each patient need to be considered, along with the age of onset, the meaning of each mood episode, its severity, and the clinical and social circumstances. Several questions remain to be answered. For instance, are there characteristics of first-episode unipolar or bipolar patients that will indicate whether extended treatment will prevent the next episode? How long should treatment be continued, and at what dose? Given the variety of treatments available for mood disorders, are there particular subsets of patients that are better suited or matched for one agent as opposed to another? What are the characteristics of bipolar patients whose depressive episodes can be treated safely with antidepressants, without risk of switching to mania? What are the costs and benefits of extended treatment, not only in terms of side effects and health service utilization, but also in terms of whether the agents we use may actually worsen the subsequent course of illness? Further questions concern the role of psychotherapy in the continuation and maintenance treatment of recurrent mood disorders. Although few clinicians would advocate psychotherapy alone for treatment of unipolar and bipolar patients at high risk for relapse and recurrence, the specific role of psychotherapy remains to be evaluated. It will be important to determine whether structured psychotherapies can provide additional benefit in terms of both episode prevention and functional outcome. This latter issue is particularly important in view of the substantial social and occupational morbidity even in remitted unipolar and bipolar patients--morbidity which is clearly significant in terms of the overall costs of mental illness. Clearly, more work needs to be done. Even the best studies indicate that a substantial proportion of patients are not helped by current treatments. PMID- 8415236 TI - Treatment resistance in unipolar depression and other disorders. Diagnostic concerns and treatment possibilities. AB - The disorders discussed in this article share common problems with treatment resistant depression both in research and clinical treatment. Lack of clear treatment-resistant definitions leads to difficulties in comparing the results of research studies. The absence of double-blind, placebo-controlled studies that match alternative treatments against each other creates confusion for the clinician who has to decide on a treatment paradigm. The future will, no doubt, see an increase in interest in treatment-resistant research and the answer to some of these questions. Although this review emphasizes the work that remains to be done before treatment resistance becomes a curiosity of the past, it also highlights the wealth of treatment options available to clinicians and their patients. As long as treatment-resistant patients are amendable to trying new therapies, hope that their depressions will lift remains. PMID- 8415237 TI - Therapeutic drug monitoring. Principles and practice. AB - TDM represents an important tool in the optimal use of selected psychiatric medications. Its role in therapy varies depending on the drug being prescribed. For some medications, such as lithium and tricyclic antidepressants, it should be a standard aspect of care. For other drugs, such as benzodiazepines, it is not helpful. Unfortunately, the use of TDM often follows an "all or none" pattern. Some clinicians rarely use it even for drugs like TCAs where it is critical to optimal care. Others use it excessively; repeating levels when it is not necessary or for drugs for which it is not necessary. It is hoped that this article has provided a basis for the thoughtful application of TDM in psychiatry as well as a review of the available data. PMID- 8415238 TI - Drug interactions in psychopharmacology. AB - The importance of psychotropic drug interactions has become increasingly evident in recent years. Although drug interactions may lead to therapeutic benefits, they also may result in diminished efficacy of drug therapy or may cause toxic or life-threatening reactions. To avoid these unwanted effects, it is important for the clinician to be aware of the basic principles that govern drug interactions. PMID- 8415239 TI - [Role of oxygen free radicals in ischemic reperfusion lesions to the myocardium]. AB - Free oxygen radicals are highly active oxygen compounds contributing to several pathological states. A possible source of free oxygen radicals during reversible ischemic heart disease and mechanisms of their toxicity to the myocardium are discussed. Despite controversial results of clinical trials with the use of antioxidants in reducing an area of necrosis in the experimental myocardial infarction, it seems that free oxygen radicals play an important role in ischemic reperfusion heart disease. Some promising results with the use of antioxidants (significant decrease in dyskinesis of ischemic left ventricular muscle and reduction of cardiac arrhythmias incidence) may be of clinical importance. PMID- 8415240 TI - [Effect of a single cooling of the entire body in the cryogenic chamber on selected hemodynamic parameters and blood serum hormone levels in healty subjects]. PMID- 8415241 TI - [Activation of blood platelets in patients after myocardial infarction]. PMID- 8415242 TI - [Evaluation of arrhythmias during the perioperative period in patients with various forms of goiter during 24-hour electrocardiographic holter monitoring]. PMID- 8415243 TI - [Effect of indomethacin and ibuprofen on blood pressure of patients treated with nifedipine or captopril]. AB - Non-steroidal anti-inflammatory agents may moderately increase blood pressure thus affecting hypotensive agents effectiveness. This study aimed at evaluating arterial blood pressure, serum 6-keto-PGF1 and aldosterone as well as plasma renin activity in hypertensive patients treated with indomethacin or ibuprofen with captopril or indomethacin or ibuprofen with nifedipine. Captopril given at the same time as indomethacin or ibuprofen did not decrease both systolic and diastolic blood pressures. However, no interaction between nifedipine administered simultaneously with indomethacin or ibuprofen was noted. Serum 6 keto-PGF1 and aldosterone as well as plasma renin activity were decreased in both groups of treated patients. Mechanism of indomethacin and ibuprofen hypertensive action seems to se related with their ability to inhibit prostacyclin biosynthesis. PMID- 8415244 TI - [Thrombolytic therapy in acute myocardial infarction hospitalized in a general internal medicine ward]. AB - Well known benefits of the thrombolytic therapy in early phases of the myocardial infarction justify such a treatment also in these hospitals in which cardiac catheterization or cardiosurgery are inaccessible. The authors discuss their two year experience with treating patients with myocardial infarction in such conditions. PMID- 8415245 TI - [Captopril treatment--2-year period of observation]. AB - Captopril was administered to 50 carefully selected patients with severe circulatory failure (18 patients classified as class III and 32 as class IV according to NYHA) in daily dose of 37.5-75 mg for two years. Patients were also given digoxin, diuretic agents and iso-dinitrosorbide. Clinical improvement increased with duration of captopril therapy. A significant improvement following the correction of therapy was achieved in 15% of patients, following one month in 28%, three months--in 70%, and after 1 and 2 years in 84% of the treated patients. All patients survived for one year, and 44--for two years (88%). Clinical improvement was manifested by: diminished of dyspnoea, edema, pulmonary and liver congestion, increase in left ventricle ejection fraction, change of disease staging by one or two NYHA classes, and reduced ventricular rate during atrial fibrillation (in 30% of patients within one year). More noticeable improvement was seen in patients with baseline ejection fraction > 40% than those with EF < 30%, in hypertensive patients than normotensive, and in patients classified to III NYHA class. Ejection fraction increased from 37.9 +/- 9.2% before the treatment to 54.6 +/- 7.7% after a two-year captopril therapy (p < .01). Captopril greatly contributes to the successful therapy of the chronic severe heart failure. PMID- 8415246 TI - [Sotalol--pharmacology, effect on hemodynamics and interactions]. PMID- 8415247 TI - [Clinical, electrocardiographic and electrophysiologic evaluation of intravenous and oral ipratropium bromide in patients with symptomatic bradycardia]. AB - Both oral and intravenous ipratropium bromide were assessed clinically, electrocardiographically and electrophysiologically in 15 patients with symptomatic bradycardia, aged between 32 and 79 years. Ipratropium bromide administered intravenously accelerated heart rate by more than 30% in the majority of patients. Electrophysiologically it normalized conduction time and sinus node response. Electrophysiological studies did not enable to foreseen the efficiency of oral form which may be useful in the treatment of symptomatic bradycardia in increased parasympathetic tone and both mild, and moderate sinus node insufficiency. PMID- 8415248 TI - [Trimetazidine therapy in atrio-cochlear disturbances of vascular origin]. AB - Therapeutic effects of TMZ treatment were evaluated in 31 patients with atrio cochlear disorders caused by an insufficiency of cerebral circulation. It was found that the results of vestibular or cochlear disorders remission was dependent on the time interval between the onset of symptoms and the beginning of trimetazidine therapy. The best results were also found in patients suffering from atrio-cochlear disorders lasting not longer than two years and due to arterial hypertension or cerebral basilar artery insufficiency. PMID- 8415249 TI - [Bepridil--a new antianginal agent with antiarrhythmic properties]. PMID- 8415250 TI - [Natriuretic peptide receptors in brain structures responsible for blood circulation and homeostasis of body fluids]. PMID- 8415251 TI - [Heart and systemic lupus erythematosus. A survey of available literature]. PMID- 8415252 TI - [Evaluation of the ST segment in ambulatory ECG monitoring with the Holter technique]. AB - Holter ECG monitoring is a valuable diagnostic technique in both cardiac arrhythmias and ischemic heart disease, known for over 30 years. Current methods of ST segment analyses require the use of computers. Myocardial ischemia is diagnosed if ST segment deviation from isoelectric axis is at least 1 mm, duration of this change is at least 1 minute, and interval is at least 1 minute. There are four ways to measure ST segment deviation from isoelectric axis: measurement of "area of given surface", mean length of ST segment, measurement of the real time of ST segment duration, and optic technique. Use of appropriate leads system is always necessary. PMID- 8415253 TI - [Exercise echocardiography]. PMID- 8415254 TI - [Biological properties of cytokines and their role in lung pathology]. AB - In the past decade, cytokines came into focus as important multifunctional mediators regulating cellular functioning and intercellular connections. Particular cytokines may both inhibit and stimulate cells playing an important role in maintaining homeostasis of the normal tissue. In the majority of pathologies not individual cytokines but their complexes are involved. Numerous studies on cytokines are mainly aimed at evaluating the role of the following cytokines in pulmonary diseases: tumour necrosis factor alpha (TNF-alpha), transforming growth factor beta (TGF-beta), insulin-like growth factor I (IGF-I), interleukin-1 (IL-1), and recently interleukin-6 (IL-6). The possible physiologic significance and role of cytokines in lung diseases are surveyed. PMID- 8415255 TI - [Five years of home oxygen therapy in Poland]. AB - Long-term oxygen therapy (LTOT) is known since the second half of the present century. In Poland it was introduced in 1986. By the end of 1991, 37 regional LTOT centers, using almost 1000 oxygen concentrators as oxygen source, were organized all over the country. To LTOT are classified two groups of patients with the advanced chronic pulmonary disease complicated with fixed respiratory failure and frequently with cor pulmonale. One group consist of patients with severe hypoxemia (PaO2 < or = 7.3 kPa, i.e. < or = 55 mm Hg) accompanied by the signs of cor pulmonale or tissue hypoxia whereas another consists of patients with moderate hypoxemia (PaO2 7.4-7.8 kPa, i.e. 55-65 mm Hg). A 5-year history of the regional LTOT centres in Poland was assessed. According to questionnaires received from 27 centres, 971 patients (662 males and 309 females), aged 59.3 years on the average, were classified to the long-term oxygen therapy between August 1, 1986 and December 31, 1991. Six hundred fifty seven patients (67.6%) has COPD, 229 (23.6%) had non-COPD pulmonary disease, and 85 patients (8.8%) other diseases. Six hundred forty four patients (66.3%) were given oxygen concentrators for severe hypoxemia and 327 patients (33.7%) for moderate hypoxemia wit co-existing signs of the cor pulmonale and tissue hypoxia. Mean time of oxygen use in 839 patients was 13.8 hours a day. Long-term oxygen therapy was ceased in 31 patients (3.2%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415256 TI - [Value of additional qualifying tests for home oxygen therapy in patients with chronic obstructive diseases and moderate hypoxemia]. AB - A value of pulmonary artery catheterization, exercise test and blood oxygenation at sleep was assessed. Such tests are considered as the additional possibility for classifying the patients with chronic obstructive disease, with moderate hypoxia, to the treatment with oxygen at home. Hemodynamic tests have shown pulmonary hypertension in 73% of patients. Clinical evaluation of the cor pulmonale seemed satisfactory. A decrease in arterial blood oxygenation during exercise below 88%, being a border value classifying to the treatment with oxygen at home, was found in 82% of patients. Marked decrease in blood oxygenation at sleep has been characteristic feature of all patients. Exercise test and oximetry at night may also valuable for classifying the patients to the treatment with oxygen under home conditions. PMID- 8415257 TI - [Evaluation of the combined action of muscarinic receptor antagonists and beta receptor agonists on reactivity of basophils isolated from blood of patients with atopic bronchial asthma]. AB - The study aimed at evaluating effects of selective M1 receptors antagonist pirenzepine-and selected beta-receptor-agonists-orciprenaline or salbutamol, given alone or in combination, on histamine release from basophils isolated from patients with atopic asthma. Histamine concentration in the cells was assayed with spectrofluorimetric technique described by Shor and modified by Scov and Norn, using anti-IgE and metacholine as liberators. It was showed, that pirenzepine inhibits histamine release caused by both anti-IgE and metacholine. In the latter case this inhibitory effect was more significant. Beta sympathicomimetics acted conversely, significantly inhibiting immunological histamine release. This effect was weaker, if metacholine was used. Combination of these agents increased protective effect of pirenzepine on histamine release induced by metacholine. PMID- 8415258 TI - [Dynamic blood viscosity in selected respiratory system diseases]. AB - The dynamic viscosity of blood was examined in 90 persons with different pulmonary diseases ill and in 56 with a normal state of health. Age was found not to have any effect on blood viscosity both in the male and female members of the control group, whereas in women aged below 40 blood viscosity was lower from that in men both before and after 40. Blood viscosity in women after 40 did not differ from blood viscosity in men, which may be explained by the effect of hormonal factors on blood viscosity in women. In the group of ill subjects, blood viscosity values were found to be increased to various degree; the highest values of that parameter were found in persons with lungs cancer. These values depended on blood protein disturbances found in that group, especially on the high concentration in blood of fibrinogen and gamma, alfa 2 and alfa 1 globulins. PMID- 8415259 TI - [Non-invasive assessment of ventricular function and cardiac arrhythmias in patients with chronic nonspecific broncho-pulmonary disease]. PMID- 8415260 TI - [Personal studies of threshold values in planimetric measurements of pulmonary pressure]. AB - Assessment of the pulmonary pressure is of importance in cardiopulmonary diseases, the most exact data are achieved with direct measurements. However, this invasive technique, which is rarely available. Therefore, the authors have elaborated the radiological technique of pulmonary pressure assessment, based on chest X-rays. Eighty patients were examined radiologically, and pulmonary pressure was evaluated planimetrically by measurements of the intermediate artery and both arteries of the anterior upper segments diameters on X-ray. These results were compared with the measurements obtained with microcatheterization technique. The authors recommend planimetric measurements for usual practice and screening of larger number of patients. PMID- 8415261 TI - [Epidemic occurrence of respiratory tract infections due to chlamydia]. AB - An analysis included 92 children aged between 3 and 7 years, 88 children aged between 3 months and 17 years and 29 their parents with diagnosed chlamydial respiratory infections. Incidence of respiratory infections in the studies pre school children was 53.2% while in family environment -87.5% in children, and 68.9% in their parents. Infection with chlamydia was found in all members in 57.8% the examined families. All children in 70.3% families were infected. PMID- 8415262 TI - [Immediate effect of hemodialysis with cuprophane membrane and acetate containing dialysis fluid on respiratory function in patients with chronic renal failure]. AB - In 20 patients with chronic renal failure on a hemodialysis (HD) programme with a cuprophane membrane and acetate-containing dialysis fluid, gasometric parameters, ventilation disturbances and peripheral blood leucocyte count were determined. A significant fall was observed of the arterial blood oxygen pressure persisting during the first hour of HD, which was parallelled by a fall of the leucocyte count. The loss of CO2 in the dialysate is attributed the major cause of hypoxemia due to alveolar hypoventilation. In 18 patients we observed a ventilation disturbances of restrictive type which were demonstrated with decreased vital capacity (VC), reduced maximal breathing capacity (MBC), increased residual volume (RV) and lower one-second forced expiratory volume (FEV1). PMID- 8415263 TI - [Catalet--vaccine against hay fever]. AB - Catalet is the Polish vaccine against pollinosis. It contains pollens of 13 species (mainly grass) and is used in the treatment of pollinosis. It is standardized in protein units. Allergen adsorption on aluminium hydroxide prolongs its absorption and decreases a number of the necessary injections. A four-year investigations in 30 patients (including 5 children) showed that Catalet tolerability is comparable with that of foreign-made preparations such as Alavac P, Desalerga F, and Allpyral-5 Grass mix. PMID- 8415264 TI - [Nedocromil in treating chronic bronchial asthma of a moderate course]. AB - Randomized, double blind, placebo controlled clinical studies aimed at evaluating the efficiency of nedocromil sodium (Tilade) in the form of metered dosimeter aerosol. Studies involved patients with moderate chronic bronchial asthma controlled with beta 2-agonists and theophylline in the form of sustained release preparations. Forty patients completed the studies. All patients were examined clinically (staging of the symptoms and doses of drugs) and spirometrically prior to and after 4 and 8 weeks of the treatment with nedocromil sodium. Statistically significant clinical improvement and spirometric improvement as well in patients treated with nedocromil sodium were noted. It may be concluded that nedocromil sodium is effective and well tolerated adjuvant therapy in the bronchial asthma. PMID- 8415265 TI - [Clinical evaluation of budesonide forte action in patients with atopic bronchial asthma]. AB - An effect of a 6-week therapy with Budesonide forte aerosol Polfa on the course of bronchial asthma has been investigated in 30 atopic asthmatics with double blind trial. The drug has been administrated in the dose of 200 micrograms four times daily and the results have been compared with placebo and foreign made analogue. Twelve patients required chronic administration of oral glucocorticosteroids in the daily dose of 9.2 mg of prednisone. Dyspnoea, cough, doses of bronchodilators and oral corticosteroids, pulmonary function and bronchial reactivity to histamine have been considered in the clinical analysis. A decrease in the severity of dyspnoea and in the doses of used bronchodilators as well as intensity of cough have been observed during the treatment with inhaled steroids. The mean daily dose of prednisone has decreased to 5.8 mg and oral corticosteroids could be withdrawn in 4 of the patients treated chronically with these drugs. Budesonide forte significantly reduced bronchoconstriction and bronchial reactivity to histamine, compared with baseline values. PMID- 8415266 TI - [Diagnostic difficulty in a case of mucoepidermoid bronchial carcinoma]. AB - A case of a 50-year male patient is presented. A cause of hemoptysis lasting for several months could not be identified in this patient. Chest X-ray and laboratory tests were normal. Temporarily atypical tubercle (Mycobacterium scrofulaceum) were seen in sputum, making proper diagnosis difficult. Further follow-up suggested cancer of the left lung. The diagnosis was ultimately confirmed intraoperatively. Histologically exceptionally malignant cancer- mucoepidermoid carcinoma--was diagnosed. PMID- 8415267 TI - [Biomarkers of metabolic disposition to lung neoplasms]. PMID- 8415268 TI - [Enzyme converting angiotensin I. Characteristics and diagnostic meaning]. PMID- 8415269 TI - Is there any room left for non-Anglo-Saxon orthopedic literature? PMID- 8415270 TI - Dislocation following THA: comparison of two acetabular component designs. AB - One hundred ninety-seven consecutive primary cemented total hip arthroplasties using 22 mm heads were evaluated for the rate of dislocation. All surgery was performed by one surgeon through a posterior approach. A Charnley type femoral component was used in each case. Patients were divided into three groups based on the acetabular component used. Group I had 60 Charnley HPW cups implanted between January 1985 and December 1986; group II had 70 Tibac cups implanted between January 1987 and August 1987; and group III had 67 Charnley HPW cups implanted between September 1987 and February 1988. The groups were similar with regards to age, sex, original diagnosis, and surgical technique. There was a total of 11 dislocations (5.6%), of which 8 (11.4%) occurred in group II (Tibac cup). Furthermore, 6 patients (3%) developed recurrent hip dislocations, 5 (7.1%) of which were from group II. Group II had a statistically significant increase in the dislocation rate (P < .05). The authors conclude that the dislocation rate with the 22 mm Tibac cup is unacceptably high and that the design of the Charnley cup affords greater stability to the artificial hip joint than the hemispherical design of the Tibac cup. PMID- 8415271 TI - Gait analysis of dysvascular below-knee and contralateral through-knee bilateral amputees: a preliminary report. AB - Four elderly peripheral vascular insufficiency below-knee amputees, average age 58, underwent contralateral through-knee amputation for gangrene. All four became household ambulators with end-weight bearing designed prosthetic sockets and four bar linkage knees. Gait analysis was performed with two AMTI (Newton, Mass) Biomechanics Force Platforms and a Watsmart Motion Monitoring System (Waterloo, Ontario). All four were observed to apparently "lock" the four-bar linkage prosthetic knee into extension during midstance and double limb support phases of gait. All subjectively felt that their through-knee limb was their more stable limb. Weight-bearing occurred during 63% of the gait cycle on the below-knee limb, 54% on the through-knee limb, and 17% in double limb support. Walking propulsion, as measured by forefoot impulse, was similar in the two limbs. The first peak of vertical force, corresponding to the elevation of the center of body weight as it passes over the weight-bearing limb, averaged 98% of body weight on the through-knee limb and only 93% on the below-knee limb. The second peak, corresponding to the kinetic energy of the falling trunk and muscle function providing linear acceleration of the center of body weight during propulsion, averaged 96% of body weight on the through-knee limb, and only 73% on the below-knee limb. Progression of the center of pressure, a qualitative measure of limb stability, was more orderly in the through-knee limbs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415272 TI - One-stage reconstruction of the nailfold. AB - Management of nailfold deformity is a difficult problem, with only seven documented techniques of reconstruction to date. We report 17 who underwent nailfold reconstruction with use of a one-stage, dorsal rotational flap with full thickness skin graft. Results show increases in range of motion and improved nail appearance. PMID- 8415273 TI - History and current application of bone transplantation. PMID- 8415274 TI - Osteoblastoma in the metaphysis of the distal femur. PMID- 8415275 TI - Seizure-induced femur fracture after total hip replacement. PMID- 8415276 TI - Infection of a total knee arthroplasty following percutaneous transluminal coronary angioplasty. PMID- 8415277 TI - Femur fracture with ipsilateral knee dislocation. PMID- 8415278 TI - Radiologic case study. Radiation-induced osteochondroma. PMID- 8415279 TI - [Symposium--reconstructive operations in injuries of the ligaments of the knee joint. Wroclaw, 9 April 1992. Proceedings]. PMID- 8415280 TI - [Alloplastic augmentation in reconstruction of the knee joint ligaments]. AB - Alloplastic augmentation of crucial ligaments of the knee is made with absorbable or non-absorbable grafts. Surgical technics is an important factor having a decisive influence on the final result. Encouraging remote results after reconstructions of the crucial ligaments with a graft taken from the ligament of the patella and allogenic augmentation show that progress in this kind of surgery should be seen in the improvement of the quality of allogenic grafts, their biocompatibility and in perfecting of the surgical technic. PMID- 8415281 TI - [Reconstruction of the posterior cruciate ligament of the knee joint by using the patellar ligament]. AB - The method of the reconstruction of the posterior crucial ligament of the knee joint with the use of the patellar ligament in 6 patients with good stability of the joint has been presented. Properly managed rehabilitating treatment effects a good result of the operative treatment. PMID- 8415282 TI - [Reconstruction of the posterior cruciate ligament with Bruckner's method. Principles of pre- and postoperative physiotherapeutic treatment]. AB - Description of a case of reconstruction of the posterior crucial ligament in the 27 year-old patient according to the method given by Bruckner with the use of a free graft from the proper ligament of the patella is presented. The knee joint was reconstructed from two incisions: anterior--medial of Payr and a short posterior--medial. The principles of pre- and postoperative physiotherapy are included. PMID- 8415283 TI - [Postoperative management after alloplasty of the cruciate ligaments]. AB - Post-operative therapeutic rehabilitation in ligamentous-capsular injuries has a great importance and for the final result it is as the very operation. We begin it from making the patient realize that the good final result can be obtained only with patient, persistence and discipline. Early therapeutic rehabilitation after surgical treatment of ligamentous-capsular injuries is possible only when the ligament was reconstructed in a motor stable way. Painless, dosed, passive motion exercises with a limited range of movements did on a mechanical splint TELOS have a great importance for the final results. PMID- 8415284 TI - [Rehabilitative treatment after surgery of the knee ligaments]. AB - Proper management of a patient begins basically before the operation of the ligament system of the knee. Before as well as after the operation the state of the muscular force of the quadriceps muscle of thigh which conditions the proper functioning of the knee should be taken into consideration. In managing rehabilitation exercises in the first phase one should concentrate on the flexion of the knee. Deficit of extension is removed only in 3 month after the operation, never, however, with the re-dressment method. Returning of the patient to work and to sport is in the 9th month. The last control takes place 2 years after the operation. In the evaluation of the stability results of the knee is more important than the range of its mobility. PMID- 8415285 TI - [Material and technical stipulations for synthetic augmentation materials]. AB - Generally used notion of "augmentation" does not present the protective effect of the ligament which has been sutured or reconstructed by the autogenic implant. "Protection" seems to be more proper notion presenting better the fulfilled function. Cruciate ligament, sutured or reconstructed by the autogenic implant from the 1/3 of the patellar ligament, is submitted to time specified healing and rebuilding processes, lasting over 1 year. Synthetic implants application creates undoubtedly protection of mentioned above rebuilding processes from functional overloading. Implants made from the resorbable materials don't fulfill the protective role for such a long period of time. That's why the non-resorbable materials implants application becomes indispensable. We keep using the polyester woven prosthesis (TREVIRA HOCHFEST) with 200 mu mesh. This kind of ligament can be strictly connected, by means of the real augmentation, with the structure it protects and can be fixed in the bones from both distal and proximal end. It is possible also to fix it following the "over the top" method in the proximal portion. For the reconstruction of ligaments with the augmentation there is a need for the synthetic prosthesis with extensibility similar to the normal cruciate ligament extensibility. In the protective plastic operation following the "over the top" method relatively stiff implants should be applied to obtain the protective "stress shield". That's why our company's offer of the cruciate ligaments implants Trevira is so varied. We use implants of 1 mm thick and of various widths: for the ligaments prosthesis--8 mm, for the so called protective plastics--5 mm and for the augmentation--3 mm of width. PMID- 8415286 TI - [Biomechanics of the knee joint]. AB - The capsular and ligamentous structures as control system of a healthy knee-joint supported by the muscular system are responsible for the rolling and gliding motion of the femoral condyles on the tibial plateau. Both the condyles and the tibial plateau have individually developed but to each other adjusted shapes and fine structures thereby. These structures consist of hyaline cartilage at their three-dimensional surfaces and of closely packed fibrils (lamina splendens) as the final gliding zone for tensile load. The orientation of the collagenous fibres can be made visible by split lines. The chondral surfaces are indirectly in contact to each other and orthogonally stressed at the particular point of contact. The indirect contact of the cartilaginous surfaces happens under interposition of the menisci. The meniscus serves to reduce and equalize the surface pressure by its own projected surface on the one hand and by maintaining of a hydraulic pressure of the synovial fluid on the other hand. Deviations of the condylar position as a result on ligamentous instabilities or ruptures with a following occurring loss of congruence, meniscal lesions or traumatic ruptures lead to a rapid discharge of the synovial fluid under load. The result is a hydraulic head loss with direct contact of the chondral surfaces under stress leading to arthrotic deformations. Severe arthrotic deformations or very much every meniscectomy produce intraarticular lumped loads resulting in a hyper physiologic chondral pressure and malnutrition thereby. Further on there develop subchondral stress concentrations (caused by the lumped loads) leading to osseous damages, too. MR-pictures can make visible these damages. Chondromalacia, fissure or even chondrolysis are arthroscopically detectable sometimes. As after-effects of deficient knee ligaments occur pathological deviations of the femoral condyles and resulting destructions of the articular surfaces under stress enormously intensified by strongly dynamic stress after ruptures of the cruciate ligaments with a too late muscular compensation. The reconstruction of cruciate ligaments is consequently required from biomechanical view to preserve the physiological position of the articular surfaces.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8415287 TI - [Technical, material and biomechanical requirements for alloplastic replacement of the anterior cruciate ligament surgical technic]. AB - Chronically instable knee-joints progressively draw to an arthrosis as a result of the irregular kinematic. Different operative techniques to replace the anterior cruciate ligament using exclusively autologous, homologous or heterologous grafts in all cases of chronic instabilities, but the primary sole suture of acute ACL injuries, too, didn't show long-term stable results in many cases. Recurrent ruptures were found first of all at the intraarticular edge of the femoral tunnel caused by permanent abrasions of the replacement. The reason why is the large angle of the ACL and each replacement respectively under motion. The question of what tissue could or should be used for renewed autogenous reconstructions in case of re-instabilities and after using bone-tendon-bone-mid third patellar-tendon-grafts isn't yet been discussed to the end. Because of the fact that there is no isometric behaviour of the complete ACL but of a few fibres only depending on the position of the knee the complete reconstruction of the ACL is absolutely impossible, but only a partial substitute. The anatomically limited intraarticular joint-space (notch) confines the replacements in their dimension and endangers voluminous reconstructions by an impending impingement especially as a result of the immediately postoperative swelling of the transplants. The use of the mid-third-patellar-tendon-graft destroys a lot of proprioceptors and causes a partly loss of the neuromuscular balance previously impaired by the loss of the ACL including its receptors. In addition to this the still existing proprioceptors of the ligamentous stumps can be definitively destroyed by big tunnels. Each autologous graft passes a long-term transformation and at best achieves to 50% of the maximum stress capacity of an original ACL after two years. Resulting from these reasons synthetic and unlimited-ly available ligaments presented themselves to be used for ACL replacements. The Trevira ligament of polyethylenetherephthalat (= TRE-VIRA HOCHFEST 730R) seems to be best qualified therefore at the present. It meets the material and technical requirements and makes allowance for all biomechanical knowledge. A modified over the-top technique by arthroscopy or mini-arthrotomy (minimized operative trauma) enables the preservation of still existing ligamentous tissue on the one hand and guarantees the immediate postoperative functional therapy on the other hand. A recurrent instability independent of cause doesn't bring about a worse starting position for the following renewed stabilization and enables corrective operative techniques including the substitute of a ruptured synthetic ligament if required. PMID- 8415288 TI - [Experimental evaluation of using polyester prosthesis of various textures for alloplasty in cruciate ligaments of the knee]. AB - In the last decade hundreds of the knee cruciate ligaments prostheses have been implanted in patients. Late follow up results of this kind of operations are under further discussion and therefore research of such implants of the cruciate ligaments are still on the way. The author has evaluated 5 kinds of polyester ACL polyester prosthesis Trevira of the same mechanical properties with a difference only in texture. He implanted them in place of primarily removed ACL in piglets of 20-30 kg and in rabbits into dorsal muscles to evaluate their biocompatibility. After 3 and 6 months period the autopsy of piglets has been performed and macroscopic image, histologic examination and X-ray image has been evaluated. The autopsy of rabbits has been performed after 14 days and 3 months. All examination described above proved the high biocompatibility of polyester prostheses. Quite significant quantitative differences in fibro-connective tissue ingrowing into the different implants of their intraarticular, intraosseous and extraarticular portion. Prostheses of loose texture have very good connective tissue ingrowing ability and seem to be more useful in cruciate ligaments replacement. Especially useful they can be in augmentation of the autogenic tissues because of their better junction with the implant, so they created functional unit. Increased amount of connective fibre tissue ingrowing the implant can be the factor of diminishing its degradation and negative influence of the knee articulation. PMID- 8415289 TI - [Clinical picture of a fresh injury of the ligamentous system of the knee]. AB - On the basis of the experience of many surgical centres advanced in the reconstruction of the knee ligaments guiding principles for our team has been worked out. In the lack of the proper devices, like arthroscopes, the authors stress the values of the clinical investigation as well as a precise examination of the instability. It leads to increase of the detectability of injuries in course of achieving experience by the surgical team. PMID- 8415290 TI - [Radiologic diagnosis of the knee ligaments]. AB - The X-rays diagnostics of the knee joint is based in a large extend on roentgenological diagnostics. All intra-articular injuries make a lot of diagnostic problems. In suspicion of an injury of a ligament system of the knee joint function films are the main examination. But NMR is the most efficient picture examination of those injuries. PMID- 8415291 TI - [Contemporary views on treatment of cruciate ligament injuries]. AB - It is commonly assumed that the anterior crucial ligament is injured most often from all ligaments of the knee. Complexity of the structure and function of the whole capsular-ligamentous system of the knee makes the operative treatment extremely difficult. In the light of the contemporary views the problems concerning the treatment of the failure of the anterior crucial ligament belong to the main orthopedic problems requiring solution. PMID- 8415292 TI - [Reconstruction of the anterior cruciate ligament of the knee joint using Eriksson's method]. AB - The reconstruction of the anterior crucial ligament of the knee joint with the method of Eriksson has been performed in 10 patients of 17 to 42 years. In some patients complicated injuries of joint structures were observed. Control examinations included 7 patients in the period of observation over 6 months after the operation. The results were evaluated according to the 100-point scale of Lysholm-Gilquist. All patients evaluated the results of the treatment as good and obtained from 84 to 91 points. PMID- 8415293 TI - [Anterior-medial rotatory instability of the knee joint. Follow up results of surgical treatment]. AB - In the years 1979-1988 65 patients (62 men and 3 women) with rotary anterior medial instability of the knee were treated surgically. The average age was 34 years. The cause of injuries (65%) were accidents during work in mines of pit coal. 52 patients were treated surgically in the period of 7 days after the accident. The remaining 13 ones were operated on after the period of 6 weeks. In the group of patients operated on immediately the most frequently applied procedure was the suture according to O'Donoughe of the injured ligaments and shift of the goose foot according to Slocum and Larson. But when the anterior crucial ligament was broken in the shape of a "brush", its reconstruction was formally made according to Jones. For patients with chronic instability extra articular plastic operations were performed i.e. shift of the semi-tendinous muscle and shift of the goose foot, as well as intra-articular--in case of injury of the anterior crucial ligament. The period of observation was from 4 to 12 years. Better results were obtained in patients operated on immediately: good result--62%, satisfactory--32% and bad--6%. For all patients with chronic instability the obtained results were only satisfactory. PMID- 8415294 TI - [Treatment of anterior instability of the knee joint with particular regard to free grafts of the specific patellar ligament]. AB - 48 patients with injuries of the anterior crucial ligament of the knee joint have been operated on. 27 patients were included the in control in the remote observation (1-3 years). In the evaluation of the results the following things were taken into consideration: feeling of stability of the knee, rest and exercise pains or lack of them, the degree of limitation of the function, lack of ability to practice sport, and the degree of improvement of the topical state in comparison with the period before the operation. As it can be seen from the obtained clinical results and the subjective evaluation the patients bore small residual anterior instability relatively well but they felt the same failure of the collateral ligament. PMID- 8415295 TI - Nevus depigmentosus systematicus with partial yellow scalp hair due to selective suppression of eumelanogenesis. AB - We report a Japanese patient with congenital hypomelanosis with a segmental pattern on the left abdomen, whorl-like pattern on the back; mosaic pattern on the chest, right abdomen, and proximal extremities; and with yellow hair on a portion of the scalp. Chemical analysis of the yellow hair revealed decreased eumelanin content, whereas the pheomelanin content was normal. PMID- 8415296 TI - Eczematous skin reaction from patch testing with aeroallergens in atopic children with and without atopic dermatitis. AB - To determine whether aeroallergens could induce eczematous lesions, 30 patients with atopic dermatitis were studied in comparison with 30 patients with respiratory atopy without atopic dermatitis. All patients were between 2 and 14 years of age. Patch testing with five aeroallergens--housedust, mite, cockroach, mold mix, and grass mix--was done on skin that was stripped by 10 applications of adhesive tape. Intradermal tests with the same antigens were done on the forearm. In 27 (90%) children with atopic dermatitis, patch testing with aeroallergens induced eczematous lesions at one or more sites. Mite, cockroach, house dust, mold mix, and grass mix caused reactions in 21 (70%), 21 (70%), 19 (63%), 15 (50%), and 13 (43%) patients, respectively. Three patients had a dermatitis flare at the antecubital and popliteal fossae during testing. Only three (10%) atopic children without atopic dermatitis had eczematous lesions, which was significantly different from children with atopic dermatitis (P < 10(-6)). Intradermal skin tests in both groups were not significantly different. This study supports previous reports that aeroallergens plays an important role in causing eczematous skin lesions. PMID- 8415297 TI - Electron microscopic studies of harlequin fetuses. AB - Four cases of harlequin fetus of various estimated gestational ages (16, 20, 21, 24 weeks) were examined by light and electron microscopy. When the epidermis was keratinized the following features were commonly found: hyperkeratosis with or without granular cells; dilated hair follicles with plugged keratin; a large number of dense or particle-cored granules in the upper malpighian layer; absence of cementsomes (lamellar bodies); large vacuoles with peripherally located laminations; large mitochondria with vesicular or membranous cristae; and early formation of the marginal band in keratinocytes and abnormal formation of the same in luminal cells of the acrosyringium. A 16-week specimen had no sign of keratinization, which made it difficult to detect these abnormalities; however, it did have large mitochondria in the keratinocytes. The mucous membrane of the lip was thickened but not keratinized. PMID- 8415298 TI - Ultrastructural study of two patients with both piebaldism and neurofibromatosis 1. AB - Piebaldism was associated with neurofibromatosis 1 (NF-1) in two patients, an association not previously reported. Dopa staining (tyrosinase) and electron microscopy were performed: no melanocytes or melanosomes were found in hypomelanotic skin of patient 2 and in the white forelock skin of patient 1; in patient 2, normal melanocytes and melanosomes were present in the white forelock epidermis but absent from the cortex, cuticles, and inner root sheath of the white forelock hair. Because these structures receive melanosomes from melanocytes in the hair bulb, it was assumed that there were no melanocytes in the hair matrix. Melanocytes and melanosomes were normal by ultrastructural criteria and in terms of their distribution in a normally pigmented macule within a hypomelanotic patch of patient 2. These and earlier report findings led to three conclusions: subtypes of piebaldism exist, including our patients showing a combination of piebaldism and NF-1; the most commonly reported subtype has no melanocytes in the white forelock and hypomelanotic skin, although microscopic islands of melanocytes may exist within hypomelanotic skin; and the ultrastructure of white forelock skin and hair of patient 2 is consistent with a mouse model of piebaldism, in which the hair follicle has no active melanocytes, but the interfollicular epidermis is normally melanized. PMID- 8415299 TI - Piebaldism in a mentally retarded girl with rare deletion of the long arm of chromosome 4. AB - A 4-year-old mentally retarded girl had congenital depigmentations of ventrolateral parts of the chest, abdomen, and legs. She also showed dysmorphic features of the head, thorax, and extremities, a pigmented ring in both irises, and a hernia of the left obliquus muscle. Cytogenetic investigations revealed deletion of chromosome 4 for the long arm segment q12-q21. The typical depigmentations, reported in four other patients with a similar chromosomal deletion, correspond with those in the autosomal dominant piebald trait. Mutations in the Kit protooncogene (mapped to the chromosome (4q11-4q12 region) have been found in patients affected with this dominant disorder. Piebaldism in children with developmental delay and dysmorphic features should alert the physician to the possibility of a deletion of the long arm of chromosome 4. PMID- 8415300 TI - Sporadic cases of Heck disease in two Polish girls: association with human papillomavirus type 13. AB - Two sporadic cases of Heck disease in Polish girls were associated with human papillomavirus 13. No other children and nobody from their surroundings had similar lesions. The course of the disease was chronic in both patients. The lesions regressed spontaneously in one girl within several years after some of the papules were excised. The second girl's warts persist after eight years. PMID- 8415301 TI - Focal epithelial hyperplasia: Heck disease. AB - Two sisters of Mexican ancestry had focal epithelial hyperplasia (FEH). The lesions on the oral mucosa of the older child were initially misinterpreted as representing sexual abuse. Microscopic evaluation of a hematoxylin and eosin stained section from a lower lip papule demonstrated the histologic features of FEH. Although human papillomavirus (HPV) type 13 and HPV32 have been most consistently present in FEH lesions, types 6, 11, 13, and 32 were not detected in the paraffin-embedded tissue specimen of our patient using an in situ hybridization technique. The lesions persisted or recurred during management using destructive modalities; subsequently, they completely resolved spontaneously. PMID- 8415302 TI - Trichoepitheliomatous infiltration of the skin simulating leprosy. AB - A 13-year-old girl had a six-year history of infiltrated erythematous plaques on the face, alopecia of the eyebrows, diffuse alopecia of the scalp, and absence of body hair. Histologically, the lesions on the face and body corresponded to trichoepitheliomas. The lesions on the face clinically simulated lepromatous leprosy. This case probably represents an entity not previously described. PMID- 8415303 TI - Chronic bullous disease of childhood and ulcerative colitis. AB - A 12-year-old girl with ulcerative colitis (UC) treated by colectomy with rectal stump preservation, salazopyrin, and systemic steroids developed persistent mouth ulcers, cutaneous targetlike lesions, papulopustules, and bullae. The clinical, histologic, and immunopathologic features were typical of chronic bullous disease of childhood (CBDC). Although resistant to combined immunosuppressive therapy with low-dose prednisolone, cyclosporin, and thalidomide, striking remission of the mucocutaneous symptoms resulted with surgical resection of the diseased rectal stump. An etiologic association between CBDC and UC is postulated. PMID- 8415304 TI - Staphylococcus aureus as a cause of perianal dermatitis. AB - Perianal dermatitis has been reported to be caused by group A beta-hemolytic Streptococcus. We present a case caused by Staphylococcus aureus. A clinical clue pointing to this organism was the presence of satellite pustules. Identifying the pathogen in perianal dermatitis is therapeutically important, as oral penicillin VK will not be effective if S. aureus is the true cause. Other streptococcal and staphylococcal cutaneous infections may exhibit overlapping clinical features, including scarlet fever, impetigo, toxic shock syndrome, and cellulitis. PMID- 8415305 TI - Hypertrichosis universalis congenita: a separate entity, or the same disease as gingival fibromatosis? AB - Hypertrichosis universalis congenita is an extremely rare disorder characterized by generalized hypertrichosis. It is generally accepted as being inherited as an autosomal dominant trait with varying expression. Many aspects of this disease are still unknown. Several reports associating hypertrichosis and gingival fibromatosis raise the question of whether they are separate entities or the same disease with different expressions of the underlying process. Hypertrichosis universalis congenita occurred in a 6-year-old girl without known family history. Her facial features were simian-like and her gingiva was moderately hyperplastic. We pose the question of whether or not these phenomena are related. PMID- 8415306 TI - Hemorrhagic complications in a rapidly growing, congenital hemangiopericytoma. AB - A 3-week-old infant had a massive hemangioma-like vascular neoplasm that had enlarged rapidly since being noted at birth. Less than a day after this initial evaluation the tumor underwent spontaneous ulceration and the infant had a near fatal hemorrhage. Histologically, the tumor was a spindle cell neoplasm most consistent with the diagnosis of congenital hemangiopericytoma. Local excision appears to have been curative after almost two years of follow-up. The case is instructive in pointing out the importance of considering nonhemangioma vascular neoplasms in the evaluation of newborns with vascular tumors. PMID- 8415307 TI - Subcutaneous fat necrosis of the newborn and hypercalcemia: case report and review of the literature. AB - Subcutaneous fat necrosis of the newborn (SCFN) alone is an uncommon condition. Its association with hypercalcemia has been reported in 19 neonates since 1926. The two occur in full-term to postterm newborns with perinatal complications associated with delivery. Erythematous to violaceous, firm, subcutaneous nodules appear approximately 1 to 4 weeks after delivery, preceding the development of signs and symptoms of hypercalcemia. Although SCFN and hypercalcemia are rare complications in neonates with perinatal problems, death due to the sequelae of hypercalcemia occurred in 3 of the 19 patients. A neonate who develops skin lesions consistent with SCFN should be followed for possible onset of hypercalcemia and treated in a timely fashion. PMID- 8415308 TI - Infantile generalized pustular psoriasis associated with lytic lesions of the bone. AB - Generalized pustular psoriasis is rare in children, especially in those less than 1 year of age. Lytic lesions of the bone have been reported in children with psoriasis, but are rare. We describe an infant with the clinical and histopathologic features of generalized pustular psoriasis that began in the first few weeks of life. In addition, this patient had sterile lytic lesions of the bone. Despite significant improvement in the bone lesions, his skin condition was resistant to therapy. PMID- 8415309 TI - Recurrent giant aphthous ulcers in a child: protracted treatment with thalidomide. AB - A child affected by giant recurrent aphthous ulcers was treated successfully over the long term with thalidomide, with no adverse reactions or reduction of therapeutic efficacy. The use of thalidomide in children for serious aphthosis is proposed. PMID- 8415310 TI - Misunderstanding of cosmetic products: a paradigmatic clinical case in a baby. AB - A 6-month-old baby had erythema, fissuring of the skin, and craquele due to prolonged use of a liquid soap mistakenly believed to be a moisturizer. We think these kinds of skin disorders will become increasingly common due to such misunderstandings. The proliferation of cosmetic products for baby skin care, the increasing prescription of cosmetics by dermatologists for skin disease, and the increase in kinds of cosmetics with similar or unusual contents all encourage such accidents. PMID- 8415311 TI - What syndrome is this? Sjogren-Larsson syndrome. AB - Sjogren-Larsson syndrome is an inborn error of metabolism with its primary clinical manifestations being congenital ichthyosis, spastic hemiplegia or quadriplegia, and moderate to severe mental retardation. Definitive diagnosis can be made by measuring fatty aldehyde dehydrogenase activity in cultured skin fibroblasts. Treatment of dermatologic manifestations is best accomplished with topical emollients and frequent bathing or showering. Some evidence supports medium-chain triglyceride diets and oral retinoids. These modalities, however, remain controversial. PMID- 8415312 TI - Pruritic pustular eruption in an infant. PMID- 8415313 TI - The Australian NHMRC Twin Registry: a resource for pediatric research. PMID- 8415314 TI - Efficacy of topical erythromycin in treatment of perianal streptococcal dermatitis. PMID- 8415315 TI - A new syndrome? PMID- 8415316 TI - Acute febrile neutrophilic dermatosis in childhood (Sweet syndrome) PMID- 8415317 TI - Nevus depigmentosus in India: experience with 50 patients. PMID- 8415319 TI - Diffuse cutaneous mastocytosis: a rare entity. PMID- 8415318 TI - Accessory nipples and urinary tract malformation. PMID- 8415320 TI - [Molecular and cellular aspects of cis-diamminedichloroplatinum (II) interactions with DNA]. AB - Cis-DDP is an antitumor drug with activity against various human tumors. It is generally accepted that DNA is the cellular target of cis-DDP. The present paper describes the binding models of cis-DDP to DNA, structural distortion in DNA induced by covalent binding of cis-DDP and repair studies of Pt-DNA adducts in pro- and eukaryotic systems. We also discuss the possible mechanisms of acquired resistance of cancerous cells to cis-DDP and analogues, and potential clinical implication of this phenomenon. PMID- 8415321 TI - [Clinical features of the genetic variants of alpha-1-proteinase inhibitor]. AB - Main variants of alpha-1-proteinase inhibitor are reviewed. Some clinical aspects of alpha-1-proteinase inhibitor deficiency concerned with emphysema, hepatic cirrhosis and panniculitis are discussed. PMID- 8415322 TI - [Lipoxins--new arachidonic acid-derived molecules]. AB - The paper deals with the current state of knowledge on lipoxins. The structure, multiple biosynthesis pathways, possible mechanisms of action, biological activities and participation of lipoxins in human pathology are characterised. PMID- 8415323 TI - [Specific inhibitors of plasminogen activators]. AB - The article contains a survey of the recent papers on the fibrinolytic inhibitors which regulate the process of plasminogen activation--PAI-1 and PAI-2. Both of them belong to serine protease inhibitors family and are specific for tPA as well as uPA, forming stable complexes in molar ratio 1:1 with them. PAI-1, secreted mainly by endothelial cells and platelets, is the main plasmic inhibitor of plasminogen activation. PAI-2, originated essentially from placenta, appears in blood during pregnancy. PMID- 8415324 TI - [Professor Henryk Matej]. PMID- 8415325 TI - [Prions]. AB - Prions are proteinaceous infectious particles. They are probably devoid of nucleic acid. Prion protein (PrP) is encoded by the normal cellular gene. Pathological isoform of prion protein has different conformation than the normal PrP protein. The exact nature of the posttranslational modification of prion protein is unknown. This modification may be caused by infection with prions (infectious disease) or occurs spontaneously (sporadic disease). Some mutations in PrP gene results in the production of pathological protein (familial disease). PMID- 8415326 TI - Antiarrhythmic therapy in patients with congestive heart failure. AB - Current information suggests that in patients with congestive heart failure (CHF) who have asymptomatic ventricular arrhythmias, coronary artery disease (dilated ischemic cardiomyopathy), and a positive signal-averaged electrocardiogram, electrophysiologic studies are useful for stratifying risk and guiding therapy. Therapy with amiodarone hydrochloride (Cordarone) appears to improve survival in patients after myocardial infarction. In CHF patients with asymptomatic ventricular arrhythmias and dilated nonischemic cardiomyopathy, electrophysiologic studies are of little value in risk stratification because of their low yield of sustained monomorphic tachycardias. There is little evidence that therapy with conventional antiarrhythmic agents improves survival. Although amiodarone can suppress ventricular ectopic beats, no trial yet conducted has detected an effect on mortality. Randomized, controlled trials with low-dose amiodarone are needed for more definitive information. Most symptomatic patients should undergo electrophysiologic testing and receive guided therapy, either with antiarrhythmic drugs or with an implantable cardioverter-defibrillator. PMID- 8415327 TI - Clinton, Griffin, Nader, and Dole do agree on some things. PMID- 8415328 TI - When the lungs are involved by connective tissue disease. AB - Pulmonary involvement by a connective tissue disease can result in clinically important complications. Pathogenic mechanisms vary from granulomatous reaction and interstitial inflammation to primary vasculitis and immune complex-mediated disease. Understanding the pulmonary complications of connective tissue diseases is challenging in that several distinct patterns of involvement are associated with the same disease but the same lung abnormalities are found with several different diseases. Early recognition and treatment of pulmonary involvement may offer the patient a better chance of recovery from serious conditions that often carry a grim prognosis if undetected. PMID- 8415329 TI - Myasthenia gravis. Diagnostic methods and control measures for a chronic disease. AB - Myasthenia gravis should be entertained as a possible diagnosis in any patient who presents with muscle weakness or visual disturbance. The most common symptom is exacerbation of muscle fatigue with repeated use of the muscle and improvement with rest. Several inexpensive diagnostic tests for myasthenia gravis that can be performed by primary care physicians are available. Standard therapy in most cases is early thymectomy, followed by a highly individualized medication program that usually includes the anticholinesterase drug pyridostigmine bromide (Mestinon, Regonol). Corticosteroids and other immunosuppressive agents (usually azathioprine [Imuran]) may be added or substituted if response to anticholinesterase therapy is inadequate. Although myasthenia gravis is a chronic disease, it can be well controlled in most patients, provided they comply with treatment. Patient education is therefore essential. PMID- 8415330 TI - Osteomyelitis. A commonsense approach to antibiotic and surgical treatment. AB - Therapy for osteomyelitis requires a multidisciplinary approach. A precise microbiologic diagnosis and adequate debridement of necrotic tissue are essential. Acute hematogenous osteomyelitis usually responds to antimicrobial therapy. The presence of an abscess or a metaphyseal cavity in hematogenous osteomyelitis and evidence of spinal cord compression in vertebral osteomyelitis require surgical treatment. Chronic osteomyelitis usually implies that dead bone is present, which requires surgical debridement. Because of the chronicity of the infection and various presentations and surgical approaches, antibiotic treatment must be individualized. In general, however, at least 4 weeks of therapy is required. PMID- 8415331 TI - Parkinson's disease camouflaging early signs of hypothyroidism. AB - Development of hypothyroidism in a patient with Parkinson's disease may be overlooked because the clinical manifestations of the two disorders are similar. In addition, drugs used to treat Parkinson's disease may mask the slight rise in thyrotropin level that is characteristic of the early stages of hypothyroidism. In this article, the authors discuss a case in which the diagnosis of hypothyroidism was delayed in a patient who had previously been diagnosed with signs and symptoms of Parkinson's disease. PMID- 8415332 TI - Prostatitis. Sorting out the different causes. AB - Both acute and chronic bacterial prostatitis are generally caused by gram negative organisms. However, acute infections are seen in younger men and cause fever, difficulty in voiding, low back and perineal pain, and other systemic symptoms. Chronic infections are more often seen in older men and may or may not be associated with symptoms of prostatitis. Nonbacterial prostatitis is the most common type. Patients have symptoms and signs of prostatitis, but infecting organisms cannot be demonstrated. Studies attempting to identify a causative organism have not been conclusive. Prostatodynia is a complex of symptoms similar to prostatitis that occurs without objective findings that definitely implicate the prostate gland. Bladder disorders such as internal sphincter dyssynergia, tension myalgia of the pelvic floor, or, at times, stress and emotional problems may be associated. Management depends on the precipitating factor. PMID- 8415333 TI - 'The dwindles'. Failure to thrive in older patients. AB - Geriatric failure to thrive has three elements: deterioration in the biological, psychological, and social domains; weight loss or undernutrition; and lack of any obvious explanation for the condition. It results from the combined effects of normal aging, malnutrition, and specific physical, social, or psychological precipitants (eg, chronic disease, dementia, medication, dysphagia, depression, social isolation). Failure to thrive can be managed with a commonsense approach by primary care physicians and healthcare providers such as social workers and dietitians; extensive referral is not necessary. The key to effective care is to identify all of the precipitants and intervene early to prevent progression. PMID- 8415334 TI - A possible hazard in labeling of generic products. PMID- 8415335 TI - Pneumonia in the elderly. Special considerations in a special population. AB - Bacterial infections of the lower respiratory tract in the elderly may not be as atypical in presentation as traditional wisdom once held. Recent studies indicate that more than one in three elderly patients have fever, cough, and leukocytosis; nevertheless, some elderly patients present with none of the features typically associated with pneumonia. An important and consistent clinical difference between younger and older patients is the broader range of bacterial respiratory pathogens found in the elderly, including gram-negative bacilli such as Haemophilus influenzae, Proteus mirabilis, and Moraxella catarrhalis. Little is gained by the initial use of narrow-spectrum antibiotic therapy, and much may be lost. Parenteral third-generation cephalosporins and oral fluoroquinolones are active against the major pathogens and can be used for empirical broad-spectrum therapy. Recent trials indicate that results are equally good with agents of either type. Perhaps a third of elderly patients with pneumonia do not require or benefit from hospitalization. The availability of excellent new broad-spectrum oral antimicrobial agents makes treatment at home or in a nursing home an attractive way to avoid the costs and many complications of hospitalization for acute care of these frail patients. PMID- 8415336 TI - Chemotherapy-induced nausea and vomiting. Easing patients' fear and discomfort with effective antiemetic regimens. AB - Patients receiving chemotherapy should be given optimal antiemetic therapy to maximize their comfort initially and to prevent development of delayed and anticipatory nausea and vomiting. Understanding the mechanisms of chemotherapy induced nausea and vomiting allows the healthcare team to design drug regimens capable of avoiding these side effects. Prevention is important, because side effects can be debilitating and sometimes dose-limiting, and up to 10% of patients refuse chemotherapy altogether to avoid them. In general, combination antiemetic therapy is preferred over single-agent therapy for chemotherapeutic regimens that produce moderate to severe adverse effects. PMID- 8415337 TI - What to try while congestive heart failure patients are still ambulatory. AB - As understanding of the mechanisms of congestive heart failure (CHF) has improved, it has become apparent that the previously applied stepped-care approach (ie, diuretic, digitalis, then vasodilator) is no longer valid. There is compelling evidence that use of vasodilators increases survival in CHF, and angiotensin-converting enzyme (ACE) inhibitors are the vasodilators of choice. Use of an ACE inhibitor in patients with New York Heart Association classes II, III, and IV CHF improves survival over that achieved with use of placebo or direct-acting vasodilators. In patients with asymptomatic left ventricular dysfunction after myocardial infarction, long-term administration of captopril improved survival and reduced morbidity and mortality from major cardiovascular events. Using an ACE inhibitor as preventive therapy in post-myocardial infarction patients without overt CHF but with evidence of muscle dysfunction (ie, left ventricular ejection fraction 40% or less) should be considered. The role of a newer vasodilator (eg, amlodipine besylate [Norvasc]) as an adjunct to therapy remains to be defined. If current theories on the pathophysiology of CHF are correct, continued interest in beta blockers is justified, especially in newer agents that have actual vasodilatory action in addition to their other beneficial properties. PMID- 8415338 TI - Advanced congestive heart failure. Inpatient treatment and selection for cardiac transplantation. AB - Establishment of an effective oral medical regimen for a patient with advanced congestive heart failure often requires hospitalization for simultaneous adjustment of volume status and vasodilation. In the hospital, intravenous therapy is begun with nitroprusside sodium (Nipride, Nitropress) and/or nitroglycerin (Nitro-Bid IV, Tridil), combined with intravenous diuretics; it is sometimes necessary to add infusions of dobutamine (Dobutrex) or dopamine hydrochloride (Dopastat, Intropin). After 48 hours of intravenous therapy, most patients can be weaned onto oral vasodilators and eventually discharged. In patients in whom further escalation of therapies is necessary, first intravenous amrinone lactate (Inocor) or milrinone lactate (Primacor) may be added, and then epinephrine or, rarely, norepinephrine (Levophed). Steps beyond that include a trial of intra-aortic balloon counterpulsation, implantation of a mechanical assist device, or use of an external centrifugal pump. Cardiac transplantation is indicated for patients in critical or unstable conditions who have no contraindications to the procedure. The patient's profile for compliance must be carefully evaluated. Reform of the selection process is needed to identify patients who, though not critically ill, will not survive without early transplantation. PMID- 8415339 TI - Travellers' diarrhoea: slow but steady progress. PMID- 8415340 TI - Timing of intervention in acute pancreatitis. AB - This review examines the appropriate timing of intervention in acute pancreatitis. In gallstone pancreatitis, it is now clear that cholecystectomy during the primary admission carries no greater risk of complications than delayed cholecystectomy and enables earlier recovery to normal activity. This course of action pre-empts a second, possibly fatal attack of acute pancreatitis. Cholecystectomy should be done after the acute phase has settled, before discharge from hospital. Patients with gallstones should now be offered endoscopic sphincterotomy within 48 hours of admission. This approach is safe, and reduces the risk of complications. When complications develop, early necrosectomy is only indicated if conservative measures fail. Delayed (> 10 days) necrosectomy is appropriate if there is evidence of sepsis, or clinical failure to improve. Pancreatic pseudocysts can often be managed expectantly; a high proportion will resolve spontaneously. After a delay of 12 weeks, persistent cysts require evaluation by endoscopic pancreatography, which gives crucial information in the choice between percutaneous or surgical drainage of the pseudocyst. A patient with pancreatitis is usually treated under the care of a surgeon, who has traditionally taken the decision on the timing of any intervention, and has performed such intervention at open operation. Recently, the development of alternative techniques has enabled the surgeon to call on the skills of his colleagues in endoscopy and interventional radiology. However, the availability of these alternatives to surgery should not affect the timing of intervention unless it can be clearly shown that such a change in timing combined with the minimally invasive technique can improve the outcome for the patient. Intervention may be required to deal with gallstones in the gallbladder or in the bile duct, to deal with, or ideally prevent, the deleterious systemic effects of pancreatic and peripancreatic necrosis, or to drain a peripancreatic abscess. Peripancreatic fluid collections and pancreatic pseudocysts may also require either internal or external drainage to relieve symptoms or prevent complications. PMID- 8415342 TI - Anterior pituitary function after adrenalectomy in patients with Cushing's syndrome. AB - We assessed anterior pituitary function in five patients with Cushing's syndrome before and after the removal of cortisol-secreting adrenal adenomas. Before surgery, all patients lacked response of growth hormone to hypoglycaemia, four had low responses of thyrotrophin to thyrotrophin releasing hormone, three had hypogonadism and two had low prolactin reserve. After successful removal of the adrenal adenoma, all patients developed postoperative hypoadrenocorticism and recovered all impaired anterior pituitary hormones within a period of 3 months. Our results point to a direct inhibiting action of glucocorticoids at the pituitary level as the explanation for the impaired anterior pituitary function. Moreover, direct gonadal suppression by glucocorticoids may be an additional mechanism of hypogonadism in some patients. PMID- 8415343 TI - The age of onset and sex distribution of insulin-dependent diabetes mellitus in Africans in South Africa. AB - Type I (insulin-dependent) diabetes mellitus (IDDM) appears to be rare in indigenous African populations, but little detailed information has been published. We have therefore analysed the age of onset of diabetes in 176 African patients with IDDM (age of onset < 35 years), and in 149 consecutive white patients of European extraction for comparison. In the Africans the peak age of onset occurred at 22-23 years (median 22 years) with an earlier peak from 14 to 17 years. In the Whites, the peak was found at 12-13 years (median 12 years). In only 7% of Africans did diabetes start under the age of 12 years. There was a slight female preponderance in the Africans, especially at the ages of greatest incident (20-25 years)-F:M 1.55:1. When patients with duration of diabetes less than 5 years only were analysed (the period during which early mortality among African patients was greatly reduced) the patterns of age distribution were similar to the total respective groups. A peak incidence in the winter months was noted. PMID- 8415344 TI - The prevalence of macrovascular disease and lipid abnormalities amongst diabetic patients in Sri Lanka. AB - The prevalence of macrovascular disease and hyperlipidaemia was examined in 500 patients with non-insulin-dependent diabetes mellitus attending a diabetic clinic in a Sri Lankan teaching hospital and 250 controls matched for age and gender. Macrovascular disease was assessed using a modified World Health Organisation questionnaire and modified Minnesota coding of electrocardiogram recordings. Twenty-one per cent of diabetic patients and 14.3% of controls had hypercholesterolaemia (P < 0.05). Macrovascular disease was present in 13.4% of diabetic patients and 8.2% of controls. Significant differences were seen in the prevalence of hypertension (15.6% vs 4.8%, P < 0.05), obesity (16.2% vs 9.7%, P < 0.05), peripheral vascular disease (5.6% vs 2%, P < 0.05) and electrocardiographic abnormalities (12% vs 6%, P < 0.05) in diabetic patients when compared to controls. Hyperlipidaemia and macrovascular disease is common in non-insulin-dependent diabetic patients in Sri Lanka and accounts for significant morbidity. PMID- 8415341 TI - Nephrology, dialysis and transplantation. PMID- 8415345 TI - The management of male breast cancer in Nigerians. AB - The outcome in 57 male patients with breast cancer has been analysed. Four patients with early disease had simple mastectomy (one case) and radical mastectomy (three cases). All four completed a 12-cycle cyclophosphamide/methotrexate/5-fluorouracil (CMF) course of chemotherapy. Two required orchiectomy after 23 and 38 months, respectively. Two patients were alive at 79 and 63 months. Fifty-three others with advanced disease had simple mastectomy (10 cases), radical mastectomy (38 cases) and no surgery (five cases), but only 33 completed chemotherapy and 11 were submitted to orchiectomy for recurrence after a mean interval of 19 months. Two patients were alive at 63 and 69 months, respectively. The overall survival rate was 7%. It is considered that in addition to late presentation (mean (SEM): 16.4 (2.1)) months and advanced disease (93%), ineffectiveness of the CMF regimen in male patients may have contributed to the poor survival rate in these Nigerian patients. PMID- 8415346 TI - Psychosocial aspects of chronic pelvic pain, with special reference to sexual abuse. A study of 164 women. AB - Patients with chronic pelvic pain attending a tertiary referral centre show certain social, developmental and psychological characteristics. Specifically, they appear to have fewer children and to report more paternal overprotection, and a trend towards low maternal care compared to normals. They also show more depression, free-floating anxiety and somatic anxiety than such populations. The levels are similar to those found in other outpatient populations presenting with migraine or irritable bowel syndrome. Hostility levels are greater than those in normal subjects. Overall the present patient population reports the same degree of childhood sexual abuse as do many other female clinic and community sample populations. Sexual abuse is unlikely to be a specific aetiological factor in the development of chronic pelvic pain though it may yet be found to be important in subsets of the population. PMID- 8415347 TI - Severe metabolic acidosis complicating massive ibuprofen overdose. AB - We report the progress of a patient who presented following the ingestion of ibuprofen in overdose. He survived despite developing an extremely severe metabolic acidosis. PMID- 8415348 TI - Notes: a suitable case for audit. AB - The increasing emphasis on management responsibilities and audit led us to investigate the state of surgical notes in our hospital. Twelve criteria were evaluated regarding information that should be contained in the notes and the absence of this important information was documented. Deficiencies were revealed in all criteria selected. Until hospital patients files are well kept and maintained, retrospective audit whether financial or medical will not be valid. A significant injection of resources is required to redress the situation. Hospitals which have not carried out detailed studies into the contents of their notes need to do so as a matter of urgency. PMID- 8415349 TI - Gas-forming infections. AB - A 89 year old woman was admitted with increasing confusion and difficulty in walking. Her left thigh was swollen, erythematous and had associated crepitus. PMID- 8415350 TI - Hypomagnesaemia and hypocalcaemia in a patient with ovarian carcinoma. AB - A patient with disseminated ovarian carcinoma presented with symptoms of hypocalcaemia secondary to hypomagnesaemia. The low serum magnesium (0.4 mmol/l) appeared to be due to renal leakage with no evidence of ureteric obstruction or hydronephrosis on intravenous urogram. Parathyroid hormone activity (serum levels and cAMP response) was normal, despite hypomagnesaemia. The patient's complaints resolved after magnesium and calcium supplementation. Hypomagnesaemia of malignancy is a rare but important complication, and is both poorly recognized and understood. PMID- 8415351 TI - Primary lymphoma of the gallbladder. AB - A case of primary lymphoma of the gallbladder is described which is rare in the medical literature. A 76 year old man presented with acute cholecystitis and septicaemia. Investigation showed a lung abscess and a gallbladder mass. The mass was thought to be an empyema and cholecystostomy was performed. Biopsy of the gallbladder wall showed high-grade B cell lymphoma. The patient unfortunately succumbed to overwhelming septicaemia in the postoperative period. Postmortem examination confirmed primary lymphoma of the gallbladder without dissemination. PMID- 8415352 TI - Systemic lambda light-chain deposition presenting with predominant cardiac involvement. AB - An 82 year old woman with suspected Bence Jones myeloma developed intractable fluid retention presumed secondary to cardiac failure. In addition she experienced angina pectoris, and required permanent cardiac pacing for symptomatic sinus bradycardia. Postmortem studies revealed prominent myocardial and renal deposits of lambda light-chains which were Congo Red negative, and had a non-fibrillar ultrastructure. Non-amyloidotic light-chain deposition is uncommon, and a rare cause of cardiac disease. Previous work regarding possible pathogenetic mechanisms, clinical and laboratory features and treatment is reviewed. PMID- 8415353 TI - Lower segment uterine scar rupture during induction of labour with vaginal prostaglandin E2. PMID- 8415354 TI - Haemophilus influenzae meningitis in an elderly patient despite treatment with oral cefuroxime. PMID- 8415355 TI - Reversible hyporeninaemic hypoaldosteronism and life-threatening cardiac dysrhythmias: the interaction of non-steroidal anti-inflammatory drugs and autonomic dysfunction. PMID- 8415356 TI - Carcinoma of the pancreas in a young male. PMID- 8415357 TI - Measurement of visual acuity by hospital physicians. PMID- 8415358 TI - Chicken antibodies: taking advantage of evolution--a review. AB - Laying hens are highly cost-effective as producers of antibodies compared with the mammals traditionally used for such production. Also, chicken antibodies have biochemical advantages over mammalian antibodies due to the phylogenetical differences between avian and mammalian species, resulting in increased sensitivity as well as decreased background in immunological assays. In contrast to mammalian antibodies, chicken antibodies do not activate the human complement system nor will they react with rheumatoid factors, human anti-mouse IgG antibodies, or bacterial and human Fc (fragment crystallizable)-receptors. Thus, chicken antibodies offer many advantages over mammalian antibodies and may replace such antibodies in the future. PMID- 8415359 TI - The use of Saccharomyces cerevisiae to suppress the effects of aflatoxicosis in broiler chicks. AB - Three hundred and sixty day-old commercial broiler chicks were assigned to a 3 x 2 factorial arrangement of treatments to examine the effects of three levels of Saccharomyces cerevisiae 0, .05, and .1% of feed, and two levels of aflatoxin, 0 and 5 ppm, singly and in combination on aflatoxicosis. Each treatment group was replicated three times. The S. cerevisiae and aflatoxin were incorporated in a standard commercial broiler ration and fed to chicks for 4 wk. Data were collected weekly on body weights, and at the end of the experiment on the relative weights of the liver, proventriculus, pancreas, and heart. Serum concentrations of the albumin, total protein, cholesterol, uric acid, triglycerides, and enzyme activities of alanine transaminase, aspartate aminotransferase, lactase dehydrogenase, and creatine phosphokinase were also evaluated. Results showed that chicks receiving aflatoxin-contaminated feed had suppressed body weight (456 g), which significantly improved (516 g) with the inclusion of .1% S. cerevisiae. The relative weights of liver (3.58%), heart (.916%), and proventriculus (.770%), which increased significantly with the addition of 5 ppm of aflatoxin, were restored to 3.00, .783, and .680%, respectively, with the dietary inclusion of .1% S. cerevisiae. The serum concentrations of albumin and total protein (.66 and 1.62 g/100 mL), which were significantly decreased by aflatoxin, were elevated to .88 and 2.24 g/100 mL, respectively, with the inclusion of .1% S. cerevisiae.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415360 TI - The effect of 5-fluorouracil treatment of chicks: a cell depletion model for the study of avian polymorphonuclear leukocytes and natural host defenses. AB - Two-week-old Leghorn chicks injected intravenously with 5-fluorouracil (5-FU) were found to have a three- to fivefold reduction (P < .05) in circulating polymorphonuclear leukocytes (PMN) 5 to 10 days later. The reduction in PMN was dose-dependent with a dosage of 200 mg/kg body weight of 5-FU, causing suppression of PMN to almost undetectable levels. Increasing the dosage of 5-FU to 400 mg/kg had no further effect on reducing the number of PMN in the circulation nor were overt clinical signs of toxicity observed. Single per os administration of 10(6) viable Salmonella enteritidis (SE) to individual groups of chicks treated or not treated with 5-FU at 2, 4, 6, 8, 10, or 12 days postinjection resulted in a two- to sixfold increase in the incidence of SE organ invasion (liver and spleen) beginning on Day 4 postinjection and continuing through Day 12 postinjection. The greatest increase in organ invasion occurred on Days 8 through 10 postinjection and was inversely proportional to the greatest reduction in the number of circulating PMN. Using 5-FU, a granulocytopenic chicken model was developed that can be used to study the defensive role of avian heterophils against infectious agents. The results from these preliminary experiments indicate that PMN play a significant role in reducing organ invasion by SE in Leghorn chicks. PMID- 8415361 TI - The effect of orally administered thyrotropin-releasing hormone on growth and carcass characteristics of seven- to ten-week-old broilers. AB - In contrast to some earlier studies with young broiler chickens, several recent studies have shown growth responses to exogenous growth hormone (GH) administration in broiler chickens of advanced age (Vasilatos-Younken et al., 1988; Scanes et al., 1990). It was therefore of interest to determine whether intermittent feeding of thyrotropin-releasing hormone (TRH), a known GH secretagogue, would elevate plasma GH of 7- to 10-wk-old broilers and alter their growth characteristics. Four replicate pens of 15 males and four pens of 15 females 7 to 10 wk of age were given access to feed containing 3.5 mg/kg of TRH for four 2-h periods each 24 h with 4-h periods of feed removal between. Control groups either were fed the basal ration on the same intermittent schedule or had continuous access to it. Growth hormone concentrations, measured 45 to 75 min after the start of TRH feeding periods, were significantly elevated in males on the 1st day of the experiment and after 7, 14, and 21 days when they were 7, 8, 9, and 10 wk of age, respectively. Concentrations in females were elevated on the 1st day of treatment, but not thereafter. Treatment with TRH had no effect on BW or relative gain of males at any time. It significantly increased relative gain of females during the 1st wk, but not thereafter. There were no treatment effects on 10-wk shank length, Pectoralis major weight, Gastrocnemius weight, or liver weight in either sex, but the percentage fat pad of males fed TRH was lower than that of controls fed intermittently. PMID- 8415362 TI - Chicken gonadotropin-releasing hormones enhance soluble and insoluble fibronectin production by granulosa cells of the domestic fowl in vitro. AB - Experiments were conducted in vitro to examine the effect of two chicken gonadotropin-releasing hormones, cGnRH-I ([Gln8]-GnRH) and cGnRH-II ([His5,Trp7,Tyr8]-GnRH), on fibronectin (soluble and insoluble) production by chicken granulosa cells isolated from the largest (F1; about 35 mm in diameter), and third largest (F3; 15 to 20 mm in diameter) preovulatory follicles as well as from a pool of immature small yellow follicles (SYF; 6 to 8 mm in diameter). The amounts of soluble fibronectin (fibronectin secreted into the incubation medium) and insoluble fibronectin (fibronectin associated with cells plus fibronectin attached to culture substratum) were quantified with a specific ELISA. Fibronectin secreted into the incubation medium (soluble fibronectin) by unstimulated cells increased with advanced stages of follicular maturation. Addition of both cGnRH-I and -II increased the amount of fibronectin secreted into the incubation medium by all follicular cell types. The amount of insoluble fibronectin in culture wells that contained unstimulated cells also increased with advanced stages of follicle development. Both cGnRH-I and -II increased the quantity of insoluble fibronectin by granulosa cells from all follicle types. Total (soluble plus insoluble) fibronectin production was elevated when cGnRH-I or -II was added to F1, F3, and SYF granulosa cells. The magnitude of cGnRH-I or II stimulation (percentage increase) of soluble, insoluble, or total fibronectin production was calculated as a multiple of the unstimulated (control) value for each follicle type, and they were greatest in cells derived from developing and immature follicles. These results indicate that homologous cGnRH-I and -II are capable of directly modulating the physiology of the avian ovary. PMID- 8415363 TI - A problem with pain relief. PMID- 8415364 TI - Fungal nail infections. PMID- 8415365 TI - Orthopaedic day surgery. PMID- 8415366 TI - Chronic low back pain. PMID- 8415367 TI - Options for referral. PMID- 8415368 TI - The business of practice. PMID- 8415369 TI - Renal medicine. PMID- 8415370 TI - Diuretics in clinical practice. PMID- 8415371 TI - Urinary tract infection. PMID- 8415372 TI - Chronic renal failure. PMID- 8415373 TI - Investigation of haematuria. PMID- 8415375 TI - Old versus new antidepressants. PMID- 8415374 TI - Prescribing within a budget. PMID- 8415376 TI - Illness after foreign travel. PMID- 8415377 TI - Vasectomy reversal by microsurgery. PMID- 8415378 TI - Teaching resuscitation techniques. PMID- 8415379 TI - A protocol for care. PMID- 8415380 TI - Eye problems. PMID- 8415381 TI - Ocular emergencies. PMID- 8415382 TI - Advances in cataract surgery. PMID- 8415383 TI - Contact lenses and their complications. PMID- 8415384 TI - The ageing eye. PMID- 8415385 TI - Anxiety and psoriasis. PMID- 8415386 TI - Tympanometry in general practice. PMID- 8415387 TI - Measuring anxiety and depression. PMID- 8415388 TI - Advances in coronary care. PMID- 8415389 TI - Asthma management. PMID- 8415390 TI - Immunisation--125 years of progress. PMID- 8415391 TI - Drugs for preterm labour. PMID- 8415392 TI - Practice formularies. PMID- 8415393 TI - Tertiary amines related to brompheniramine: preferred conformations for N oxygenation by the hog liver flavin-containing monooxygenase. AB - The metabolism of racemic, (D)- and (L)-brompheniramine, a widely used antihistamine, was studied with microsomes and with highly purified flavin containing monooxygenase (FMO) from hog liver. In addition, a number of other similar tertiary amines were evaluated as substrates for FMO activity from hog liver and the kinetic constants obtained were compared with brompheniramine. Although some N-demethylation was observed, the major metabolite of brompheniramine and the other tertiary amines examined in hog liver microsomes was the metabolite containing an aliphatic nitrogen N-oxide. Brompheniramine was extensively N-oxygenated by the highly purified FMO from hog liver. N-Oxygenation of brompheniramine in both microsomes and with highly purified FMO from hog liver was enantioselective. The Km for N-oxygenation of (D)-brompheniramine was markedly lower than the Km for (L)-brompheniramine. (E)- and (Z)-zimeldine are less conformationally flexible model compounds of brompheniramine, and these compounds were also examined and were found to be stereoselectively N-oxygenated by the highly purified FMO from hog liver. The similarities and differences in Km and Vmax values were evaluated in terms of possible conformations of the substrates determined by SYBYL molecular mechanics calculations. Distance map data indicated that FMO preferentially accommodated selected conformations of tertiary amines. Thus, (D)-brompheniramine and (Z)-zimeldine presumably have the aliphatic tertiary amine nitrogen atom and aromatic ring center at a defined distance and geometry and were more efficiently N-oxygenated than their respective isomers. PMID- 8415394 TI - Proteolysis of human growth hormone by rat thyroid gland in vitro: application of electrospray mass spectrometry and N-terminal sequencing to elucidate a metabolic pathway. AB - The present studies were designed to provide structural characterization of peptide metabolites of biosynthetic human growth hormone (hGH) formed by rat thyroid gland proteases in vitro. Electrospray ionization mass/spectrometry (ESI MS) and N-terminal sequencing were used to characterize the peptide metabolites. The predominant enzyme in the thyroid gland preparations was a chymotrypsin-like serine protease which was biochemically similar to rat mast cell protease-I. Metabolic intermediates were formed by cleavage of hGH exclusively at Tyr/Phe/Leu Xaa bonds. After a 5- or 45-min incubation of hGH with thyroid gland S9 pellet fraction, the majority of metabolites formed were two-chain variants of hGH having masses ranging from 16,002 to 22,143 Da. These metabolites were formed as a result of proteolysis in the large disulfide loop region of hGH in combination with processing at Tyr42-Ser43. Based upon the time-related appearance and structural characterization of these intermediates, a sequence of metabolic events is proposed. The initial event appears to be cleavage by the chymotrypsin like protease between Tyr143-Ser144 to form a two-chain hGH. This intermediate is then cleaved between Tyr42-Ser43, liberating the N-terminal peptide, Phe1-Tyr42. Two other metabolites were generated as a result of the deletion of the peptides Lys140-Tyr143 and Ser144-Phe146 from the large loop region. The identification of similar metabolites truncated by a single amino acid at the carboxyl terminus indicated the action of a carboxypeptidase on these metabolic products. After a 4.5-hr incubation, the protease isolated from the S9 pellet fraction degraded hGH to > 20 small peptides, having masses < or = 2300 Da.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415395 TI - Gelatin-acacia microcapsules for trapping micro oil droplets containing lipophilic drugs and ready disintegration in the gastrointestinal tract. AB - Nonhardened gelatin-acacia microcapsules were studied for encapsulation of microdroplets of oil solution containing a lipophilic drug as core material and ready disintegration with release of micro oil droplets in the gastrointestinal tract. Probucol and S-312-d, a Ca-channel blocker, were employed as model lipophilic drugs. Glyceryl tricaprylate and tricaprate mixture solutions containing these drugs were encapsulated according to the complex coacervation method and were recovered as free-flowing powders without any hardening (cross linking) step. The microcapsules obtained were disintegrated, and the emulsion was reproduced within 3 min at 37 degrees C in the first or second test solution defined in the Japanese Pharmacopeia XII. When the microcapsules were stored as a powder at room temperature in a closed bottle, no significant change in their appearance or disintegration time upon rehydration was observed even after 1 year. Oral bioavailabilities of model drugs from the microcapsules were tested in rats and dogs and compared with those from other conventional formulations. Gastrointestinal absorption of both probucol and S-312-d from the microcapsules was remarkably more efficient than that from other formulations such as powders, granules, or oil solution. The proposed method for microencapsulation could be useful for powdering drug-containing oil solutions or O/W emulsions while maintaining excellent bioavailability. PMID- 8415396 TI - Selective paracellular permeability in two models of intestinal absorption: cultured monolayers of human intestinal epithelial cells and rat intestinal segments. AB - New data on the permeabilities of hydrophilic markers in two commonly used in vitro models, i.e., excised intestinal segments from the rat and monolayers of Caco-2 cells, are presented. The results are compared to human in vivo data. Two groups of hydrophilic marker molecules were tested: (1) monodisperse polyethylene glycols of molecular weights ranging from 194 to 502 g/mol and (2) a heterogeneous group of molecules consisting of urea, creatinine, erythritol, and mannitol (60-182 g/mol). The permeabilities of the marker molecules showed a nonlinear dependence on the molecular weight and decreased in the order rat ileum > rat colon > Caco-2 cells. Surprisingly, the polyethylene glycols permeated more easily than the other marker molecules, indicating that characteristics other than molecular weight, e.g., the flexibility of the structure, may also be important for permeation through the membrane. Comparisons with the published permeability profiles of polyethylene glycols in human intestinal segments in vivo (i.e., calculated permeability coefficients as a function of molecular weight) indicate that the human intestine is more permeable than the in vitro models. However, the permeability profiles of the corresponding segments in the human intestine and the in vitro models were comparable. Thus, good correlations were established between permeabilities of the human ileum and rat ileum and between those of human colon, rat colon, and the Caco-2 cells. We conclude that the paracellular absorption in humans can be studied mechanistically in these in vitro models. PMID- 8415397 TI - Liposome-mediated delivery of gallium to macrophage-like cells in vitro: demonstration of a transferrin-independent route for intracellular delivery of metal ions. AB - Gallium (Ga) prevents the activation of macrophages and might be useful as an immunosuppressive agent. It is taken up by the malignant cells through the transferrin (Tf) receptor pathway, but this pathway may be insufficient in the case of non-malignant cells. We studied the Tf-independent, liposome-mediated delivery of Ga to macrophage-like cells in vitro by a growth inhibition assay. The growth inhibitory properties of Ga for other types of cells was also evaluated. Ga complexed with nitrilotriacetate (GaNTA) and encapsulated in DSPG liposomes was 16 and 48 times more potent for RAW 264 cells than free GaNTA and Ga-nitrate, respectively. CV1-P cells were also somewhat sensitive to liposomal Ga, but other cell lines with lower endocytotic capacity were insensitive. The inhibition of RAW 264 cell growth induced by liposomal or free GaNTA was partially reversed with iron-loading of the cells, indicating that this form of Ga causes an intracellular iron deficiency similar to that produced by Tf-bound Ga. Our results indicate that encapsulation of Ga in negatively charged liposomes provides a transferrin independent route for intracellular delivery of the compound to macrophages, which is of special interest in the treatment of autoimmune diseases, such as rheumatoid arthritis. PMID- 8415398 TI - Solubilization of thiazolobenzimidazole using a combination of pH adjustment and complexation with 2-hydroxypropyl-beta-cyclodextrin. AB - The thiazolobenzimidazole 1-(2,6-difluorophenyl)-1H,3H-thiazolo[3,4-a] benzimidazole, TBI, is an experimental drug for the treatment of AIDS which exhibits a low water solubility (11 micrograms/mL) and is therefore difficult to administer in an injectable solution dosage form at a target solution concentration of 10 mg/mL. The compound has a single ionizable functional group and exhibits an increase in solubility with decreasing pH consistent with a pKa of 3.55, but the maximum solubility attainable by pH adjustment has been shown to be only 0.4 mg/mL (at pH 2). TBI has been found to form inclusion complexes in either its neutral or its protonated form with 2-hydroxypropyl-beta-cyclodextrin (HPCD). The equilibrium constants for 1:1 complex formation were found to be 81 and 1033 M-1 for the protonated and neutral species, respectively. Although the formation of protonated complex is less favored in comparison to the neutral complex, the contribution of this species to the overall solubility of TBI predominates at low pH. Thus, using a combined approach of pH adjustment and complexation with HPCD, a solubility enhancement of 3 orders of magnitude is possible. NMR proton spectroscopy and molecular modeling studies, conducted to understand the orientation of TBI in the complex and the effect of protonation, are described. PMID- 8415399 TI - Programmable drug delivery from an erodible association polymer system. AB - An erodible association polymer system based on blends of cellulose acetate phthalate (CAP) and Pluronic F127, a block copolymer of poly(ethylene oxide) and poly(propylene oxide), has been investigated for its applicability to rate programmed drug delivery. The compatibility and thermal properties were characterized by DSC and FTIR. Results from the thermal analysis indicate that the blends are compatible above 50% CAP, as revealed by a single composition dependent glass transition temperature (Tg). The existence of molecular association through intermolecular hydrogen bonding between the carboxylic acid and the ether oxygen groups is supported by the observation of an upward shift in the IR carbonyl stretching frequency at increasing Pluronic F127 concentrations. Using theophylline as a model drug, the in vitro polymer erosion and drug release characteristics of the present polymer system were evaluated at different buffer pH's on a rotating-disk apparatus. The results show that the rates of both polymer erosion and drug release increase with the Pluronic F127 concentration in the blend. Further, at pH 4, the polymer erosion is minimal and the theophylline release appears to be governed mainly by diffusion through the polymer matrix. In contrast, at pH 7.4, the theophylline release is controlled primarily by the polymer surface erosion. To demonstrate the unique approach to programmed drug release based on the concept of nonuniform initial drug distribution, pulsatile patterns of drug release have been achieved successfully from the present surface erodible polymer system using a multilaminate sample design with alternating drug loaded layers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415400 TI - Characterization of oil-in-water emulsions prepared from solid-state emulsions: effect of matrix and oil phase. AB - Emulsions (o/w) were prepared from solid-state emulsions comprised of various matrix materials and oils and the resultant particle size properties determined. Results suggest that for those matrices that can form solid-state emulsions, the droplet size decreased as a function of time, as previously observed. The final droplet size was dependent on the oil utilized but was independent of the matrix material. The use of mineral oil resulted in the smallest droplet diameter (approximately 1.5 microns) while isopropyl myristate resulted in the largest droplet diameter (approximately 3 microns). With the exception of mineral oil, the oil/water interfacial tension was found to be directly proportional to the droplet diameter. The rate of emulsification appeared to be biphasic. The initial emulsification phase appeared to be independent of the matrix material while the terminal phase was a function of the matrix material. Most importantly, it was found that solid state emulsions could be prepared from a diverse, yet specific, list of matrices. PMID- 8415401 TI - Biopharmaceutics of didanosine in humans and in a model for acid-labile drugs, the pentagastrin-pretreated dog. AB - Didanosine is a purine nucleoside analogue approved for the treatment of human immunodeficiency virus infection. It is extremely unstable at pH values less than 3 and requires protection against gastric acid-induced hydrolysis. Beagle dogs pretreated with pentagastrin, an analogue of gastrin that reproducibly stimulates gastric acid secretion, have been used to screen different didanosine formulations. The absolute bioavailability of didanosine from a saline solution decreased from approximately 43% in untreated dogs to 8% after pretreatment with pentagastrin. Administration of buffered solution of didanosine to untreated and pretreated dogs yielded bioavailability estimates of 37 and 30%, respectively. In humans, the bioavailability from a similar buffered solution was approximately 40%. Pentagastrin-pretreated dogs were used to evaluate four new products relative to a citrate-phosphate buffer sachet, the formulation selected for large scale clinical trials in humans. Two of these new formulations, a chewable tablet and an antacid suspension, were more bioavailable then the reference sachet. This also proved to be true in man, necessitating an adjustment in the dose of didanosine when administered as the chewable tablet. Dogs pretreated with pentagastrin accurately predicted the improved bioavailability of new didanosine formulations prior to clinical use. This animal model may be helpful in evaluating the biopharmaceutics of other acid-labile drugs. PMID- 8415402 TI - Pharmacokinetics of diltiazem and its metabolites in dogs after oral administration of a multiparticulate sustained-release preparation. AB - Pharmacokinetics of diltiazem and its six metabolites were compared after oral administration in dogs of a multiparticulate sustained-release diltiazem preparation (HER-SR, QD) and a conventional diltiazem preparation (HER, TID). The plasma concentration of diltiazem, its two active basic metabolites (M1, N monodesmethyl diltiazem; M2, deacetyl diltiazem), and four acidic metabolites [A1, (+)-(2S,3S)-2-(4-methoxyphenyl)-3-acetoxy-4-oxo-2,3,4,5,-tetrahydro-1,5- benzothiazepin-5-acetic acid; A2, 3-deacetyl-A1; A3, O-demethyl-A1; A4, O demethyl-3-deacetyl-A1] following several administration routes were determined using high-performance liquid chromatography with UV detector (UV-HPLC). Following the oral administration of HER to dogs, plasma concentrations were in the descending order of A2, diltiazem, M1, and M2. The absolute bioavailability of diltiazem was about 30%. Diltiazem conversion to its metabolites (M1, M2, A2) was 31.0, 2.1, and 14.6%, respectively. Following intraduodenal and mesenteric venous administration of diltiazem, M1 and A2 were produced mainly in the intestine and liver. Oral administration of HER-SR and HER to dogs resulted in almost-identical plasma concentrations of A2, diltiazem, M1, and M2 (descending order). Supported evidence was the effective absorption of diltiazem from all gastrointestinal tract regions and similar formation ratios of diltiazem basic metabolites (M1, M2) from the duodenum, ileum, and colon. PMID- 8415403 TI - Stereoselective interactions of organic cations with the organic cation transporter in OK cells. AB - Recent studies have suggested that certain organic cations, such as pindolol and the diastereomers, quinine and quinidine, may be stereoselectively secreted by the kidney in humans. The goal of this study was to determine if the enantiomers of pindolol, verapamil, and disopyramide and the diastereomers, quinine and quinidine, interact stereoselectively with the organic cation transporter in the brush border membrane of the opossum kidney cell line. All organic cations tested inhibited the uptake of tetraethylammonium (TEA). The IC50 values (mean +/- SD) were as follows: quinine (17 +/- 2 microM). quinidine (51 +/- 13 microM), S-(-) pindolol (23 +/- 4 microM), R-(+)-pindolol (30 +/- 4 microM), S-(-)-verapamil (0.4 +/- 0.04 microM), R-(+)-verapamil (7 +/- 2 microM), R-(-)-disopyramide (27 +/- 4 microM), and S-(+)-disopyramide (66 +/- 12 microM). Each individual organic cation pair showed significant stereoselective differences in their IC50 values, with quinine, S-(-)-pindolol, S-(-)-verapamil, and R-(-)-disopyramide being the more potent species. Both enantiomers of pindolol, quinine, and quinidine appear to exhibit simple competitive inhibition of TEA uptake based upon a derived slope similar to 1.0, using a sigmoidal inhibition model. The enantiomers of verapamil and disopyramide exhibited a slope of much less than 1.0, suggesting a more complex interaction of these organic cations with the TEA transporter. Our results suggest that organic cations stereoselectively interact with the organic cation transporter in the brush border membrane of OK cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415404 TI - Demethylation kinetics of aspartame and L-phenylalanine methyl ester in aqueous solution. AB - The kinetics of demethylation of aspartame and L-phenylalanine methyl ester were studied in aqueous solution at 25 degrees C over the pH range 0.27-11.5. The pseudo-first-order rate constant for aspartame was resolved into individual contributions from methyl ester hydrolysis and diketopiperazine formation. pH rate profiles were quantitatively described by chemically reasonable kinetic schemes. Aspartame is maximally stable at pH 4 (t90 = 53 days at 25 degrees C); phenylalanine methyl ester, at pH 3. The potentiometrically measured pKa values were pKa1 3.19 and pKa2 7.87 for aspartame and pKa 7.11 for phenylalanine methyl ester. PMID- 8415405 TI - Phenytoin-lipid conjugates: chemical, plasma esterase-mediated, and pancreatic lipase-mediated hydrolysis in vitro. AB - Phenytoin-lipid conjugates obtained by covalent binding of hydroxymethylphenytoin to diacylglycerides and to 3-acyloxy-2-acyl-oxymethylpropionic acids formed dispersions with a particle size of 10-200 microns when briefly sonicated in a sodium taurodeoxycholate-containing ethanol-water mixture. In contrast to the corresponding bis-deacyl derivatives, the lipids were not significantly hydrolyzed in aqueous buffers and in plasma. Incubation with pancreatic lipase yielded primarily the bis-deacyl compounds, which are comparable to monoglycerides, and subsequently liberated phenytoin. The glyceride-derived prodrugs were better substrates for the enzyme than the 3-acyloxy-2-acyloxymethyl propionic acid derivatives. It is concluded that the phenytoin lipid conjugates are hydrolyzed by pancreatic lipase in a similar manner as natural triglycerides. PMID- 8415406 TI - Substituted xanthones as selective and reversible monoamine oxidase A (MAO-A) inhibitors. AB - 1,3-Dihydroxy-2-methylxanthone (X1), its 4-chloro and 4-bromo derivatives (X1-Cl and X1-Br), and 1,3-dihydroxy-4-methylxanthone were investigated for their inhibition activities toward MAO. A hyperbolic function was derived to fit the data and to calculate IC50 values. The compounds proved to be reversible and selective inhibitors of MAO-A, with X1 displaying the highest activity (IC50 = 3.7 microM). PMID- 8415407 TI - Synthesis and evaluation of morpholinoalkyl ester prodrugs of indomethacin and naproxen. AB - Morpholinoalkyl esters (HCl salts) of naproxen 1 and indomethacin 3 were synthesized and evaluated in vitro and in vivo for their potential use as prodrugs for oral delivery. Prodrugs were freely soluble in simulated gastric fluid (SGF) and pH 7.4 phosphate buffer and showed a minimum of a 2000-fold increase in solubility over the parent drugs. All prodrugs were more lipophilic than parent drugs as indicated by n-octanol/pH 7.4 buffer partition coefficients but less lipophilic in terms of n-octanol/SGF partition coefficients. Potentiometrically determined pKa's for prodrugs were in the range of 6.89 to 8.62 at 25 degrees C. All prodrugs were quantitatively hydrolyzed to their respective parent drugs by enzymatic and/or by chemical means. An increase in carbon chain length rendered the prodrugs more stable at pH 7.4 but less stable in SGF. The esters were generally found to be hydrolyzed rapidly in rat plasma at 37 degrees C, the half-lives being in the range of 1.2-31.0 min. Based on in vitro results, prodrugs 2c and 4c were chosen to evaluate solid-state stability, in vivo bioavailability, and ulcerogenicity. At elevated temperatures, the solid state decomposition of 2c and 4c followed biphasic kinetics, with rapid decomposition occurring initially. The prodrugs were 30-36% more bioavailable orally than the parent drugs following a single equimolar solution dose in rats. Prodrugs 2c and 4c were significantly less irritating to gastric mucosa than parent drugs following single-dose and chronic oral administration in rats. PMID- 8415408 TI - Conformational analysis of the opioid phenylmorphan and its 9 alpha-methyl analogue in solution using high-resolution nuclear magnetic resonance spectroscopy. AB - The solution conformations of the opioid phenylmorphan (5-m-hydroxyphenyl-2 methylmorphan) and its 9 alpha-methyl analogue were studied using one- and two dimensional high resolution NMR techniques. The NMR spectra were analyzed by interpreting the phase-sensitive 2-D COSY and double quantum filtered COSY spectra, 1H-1H vicinal coupling constants, and nuclear Overhauser effects in the phase-sensitive 2-D NOESY spectra. The results show that, for both compounds, a chair-chair conformation of the cyclohexane and piperidine rings is exclusively preferred with some distortion of the rings from perfectly staggered chairs. For phenylmorphans, the phenyl ring is oriented to fit into the cleft formed by the cyclohexane and piperidine rings. Thus, for the (+)-enantiomer, the phenyl group assumes the same orientation with regard to the piperidine ring as morphine consistent with the morphine-like properties of the compound. For the 9 alpha methyl analogue, the plane of the phenyl ring essentially bisects the piperidine ring to which it is attached and is outside of the required range of opioid agonists. This is consistent with the atypical properties of the two enantiomers. The NMR results are compared to the conformations of (-)-phenylmorphan and the (+)-9 alpha-methyl analogue in the crystal state and to the results of molecular mechanics (MM2) studies. PMID- 8415409 TI - The hypolipidemic effects of 1-acetyl-4-phenyl-1,2,4-triazolidine-3,5-dione in rodents. AB - 1-Acetyl-4-phenyl-1,2,4-triazolidine,5-dione (APTD), a potent hypolipidemic agent, lowered both serum cholesterol and triglyceride levels in normo- and hyperlipidemic rats at 10 or 20 mg/kg/day. The agent effectively lowered VLDL cholesterol (VLDL-C) and LDL-C content and raised HDL-C content in normal and hyperlipidemic rats treated from 4 to 8 weeks. Similar effects on the incorporation of cholesterol into the lipoprotein fractions were observed after drug treatment. Tissue lipids, e.g. cholesterol, were lowered, whereas fecal cholesterol levels were increased. APTD's primary targets were acyl CoA cholesterol acyl transferase (ACAT) for cholesterol ester synthesis and sn glycerol-3-phosphate acyl transferase (GPAT) and phosphatidylate phosphohydrolase (PPH) for triglyceride synthesis. PMID- 8415410 TI - Evaluation of moisture sorption by tablet cores containing superdisintegrants during the aqueous film coating process. AB - A physical-chemical analysis of the extent of sorption of water by tablets containing superdisintegrants was carried out following the aqueous film coating of formulated tablets. Characterization of the uncoated and coated tablet properties was conducted using thermogravimetric analysis, differential scanning calorimetry, mercury intrusion porosimetry, and measurement of the tablet tensile strength. Tablet residual moisture content, pore system characteristics, tensile strength, and glass transition temperature of the amorphous polymer components of the tablet matrix were significantly affected after the coating operation. These findings were attributed to the penetration of water from the aqueous film coating solution into the tablet matrix. PMID- 8415411 TI - Dissolution, stability, and morphological properties of conventional and multiphase poly(DL-lactic-co-glycolic acid) microspheres containing water-soluble compounds. AB - Multiphase microspheres of poly(DL-lactic-co-glycolic acid) (PLGA) containing water-soluble compounds were prepared by a multiple-emulsion solvent evaporation technique. These compounds were dissolved in the aqueous phase of a W/O emulsion with soybean oil as the oil phase. This emulsion was dispersed throughout the matrix of the microsphere. The morphological properties of the multiphase microspheres during in vitro dissolution studies were compared to those of conventional microspheres prepared from the same polymer. Drug release from the multiphase microspheres was characterized by an initial uniform release for the first 20 days followed by a more rapid phase of drug release. Chlorpheniramine maleate (CPM) and brilliant blue (BB) were the soluble model compounds investigated. The release rates of these agents from the multiphase microspheres were independent of the drug content in the microspheres. The release profiles from the conventional microspheres showed a lag time of 10 and 16 days for the CPM and BB, respectively. The dissolution rate of the model soluble compounds from the conventional microspheres increased as the loading in the microspheres increased. No differences in the degradation rate of the PLGA from the multiphase and the conventional microspheres were seen during the dissolution studies. PMID- 8415412 TI - Stability of liposomes in vitro and their uptake by rat Peyer's patches following oral administration. AB - To evaluate the usefulness of liposomes as a carrier for the targeted delivery of antigens to gut-associated lymphoid tissue, liposomal stability and uptake by rat Peyer's patches were investigated. Liposomes composed of distearoylphosphatidylcholine, phosphatidylserine, and cholesterol (DSPC liposome), or dipalmitoylphosphatidylcholine, phosphatidylserine, and cholesterol were stable in acidic solution (pH 2.0), diluted bile, and pancreatin solution. Following the oral administration of liposomes to rats, rhodamine B-PE incorporated in the lipid phase of DSPC-liposomes was preferentially taken up by Peyer's patches in the lower ileum. The uptake of rhodamine B-PE from DSPC liposomes larger than 374 nm in mean diameter was high. Orally administered DSPC liposomes of a large diameter thus appear to serve effectively as a vehicle for delivering antigens to Peyer's patches. PMID- 8415413 TI - Decreased protein-stabilizing effects of cryoprotectants due to crystallization. AB - The stabilizing effects of various additives against inactivation of an enzyme (beta-galactosidase from Aspergillus oryzae) during freeze-drying were studied, with a focus on their crystallinity. The crystalline morphology of mannitol and inositol in freeze-dried cakes depended on the solute concentrations before freezing and the freeze-drying method used. The additives in their amorphous state showed concentration-dependent stabilization of the enzyme, whereas additive crystallization during freeze-drying decreased their effects. Heat treatment before freeze-drying also caused crystallization and diminished the stabilizing effects. Noncovalent soluble aggregates were observed in the inactivated enzyme solution. These results show the importance of maintaining the amorphous state of additives used as stabilizing agents during freeze-drying. PMID- 8415414 TI - Transforming growth factor-alpha (TGF-alpha) in a semisolid dosage form: preservative and vehicle selection. AB - The selection of an ideal semisolid vehicle for growth factors presents a challenge. Some antimicrobial agents are known to delay wound healing. The objective of this investigation was to identify appropriate preservatives and vehicles for TGF-alpha. Criteria for acceptance are noninterference with the mitogenic activity of TGF-alpha as well as adequate product preservation. Vehicles considered were o/w creams, ointments, and a gel. Combinations of six preservatives were tested. Selection was determined using both microbial preservative challenge and TGF-alpha mitogenic assay. In the former, 10 species of microorganisms were inoculated into formulation samples. At selected time intervals, it was determined whether colonies decreased, increased, or remained constant. In the mitogenic assay, samples of either preservatives or formulation prototypes were introduced to TGF-alpha-stimulated fibroblast cell cultures. Mitogenesis was determined by measuring 3H-dThd uptake into newly synthesized DNA. As preservatives, sorbic acid and quaternium-15 appear to satisfy both selection criteria. A thermosetting gel appears most promising as vehicle. PMID- 8415415 TI - Oral immunization of rats with proteinoid microspheres encapsulating influenza virus antigens. AB - Influenza virus antigen microspheres were prepared by a pH-dependent process using a protein-like polymer (proteinoid) made by thermal condensation of amino acids. The efficacy of these preparations to induce specific IgG responses when used as oral vaccines in rats was evaluated. A single enteric dose of M1 entrapped in proteinoid microspheres was able to induce a significant IgG response to M1 as early as 2 weeks postdosing, while rats dosed orally with the same M1 total dose (no microspheres) showed no detectable antibody response. An unencapsulated hemagglutinin and neuraminidase (HA-NA) preparation induced a moderate anti HA-NA IgG response. A single enteric dose of HA-NA spheres induced a response in 33% of the rats; this response was up to eight times higher than that observed in the rats dosed with unencapsulated antigen. PMID- 8415416 TI - The use of statistical indices to gauge the mixing efficiency of a conical screening mill. PMID- 8415417 TI - Factors and conditions affecting the glucuronidation of oxazepam. AB - The aim of the present work was to investigate the impact of disease states and environmental and host factors on the glucuronidation of oxazepam. Glucuronidation represents quantitatively one of the most important metabolic conjugation pathways (phase II) in man for the inactivation and detoxication of xenobiotics and endogenous compounds and the liver is the major site for it to take place. Far less attention has been paid to the conjugation reactions in previous clinical research in this field compared to the immense interest in the oxidative biotransformation pathways (phase I). This fact is mainly due to the latter giving rise to active or reactive metabolites with a toxicological potential. The metabolism of oxazepam expresses exclusively the capacity for glucuronide formation. It was a prerequisite to establish the bioavailability of oxazepam prior to succeeding studies on the oral disposition of the drug. A preparation for intravenous administration was created. Clearance was chosen as measurement of the capacity to glucuronidate oxazepam. Severe decompensated liver disease was associated with a significant decrease in oxazepam clearance, that became even more obvious when corrected for by a diminished binding to plasma proteins. This increase in free fraction of oxazepam was substantial and could mainly be accounted for by low plasma albumin values. The results are in part a settlement with earlier studies on glucuronidation in liver disease and they may undoubtedly be ascribed to the severe degree of liver disease. For the first time it was shown that hypothyroidism led to a decline in the clearance and metabolism of oxazepam and paracetamol that is mainly biotransformed by glucuronidation. It was concluded that the enzymes responsible for glucuronidation in hypothyroidism are under the influence of thyroid hormones as is the case with oxidative enzymes. Further studies focused on the effect of host and environmental factors on glucuronidation. A commercially available very low calorie product for the treatment of obesity resulted in a decrease in oxazepam clearance and a lack of co-factors as a consequence of the low calorie intake was explanatorily proposed. Beta-adrenoceptor antagonists are often prescribed together with other drugs and close knowledge on interactions is mandatory but insufficient in regard of drugs being glucuronidated. Despite the mutual metabolic pathway labetalol exerted no dispositional alterations concerning oxazepam. It was moreover suggested that very elderly subjects between the age of 80 to 94 years had a reduced clearance of oxazepam.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8415418 TI - Fetal renal artery flow velocity waveforms in the presence of congenital renal tract anomalies. AB - Colour Doppler flow mapping of the renal arteries and subsequent pulsed Doppler measurement of impedance to flow in these vessels were attempted in 33 fetuses with postnatally confirmed renal pathology. The majority presented with unilateral or bilateral hydronephrosis (n = 21) and bilateral renal agenesis (n = 8). Renal artery blood flow could be visualized in all, except for the eight cases of bilateral renal agenesis. Bilateral flow velocity recordings were collected in six out of 12 cases of bilateral hydronephrosis and in five out of nine cases of unilateral hydronephrosis. The pulsatility index (PI), as a measure of downstream impedance, was in the normal range in 16 out of 18 kidneys (88 per cent) in bilateral hydronephrosis and in 12 out of 14 kidneys (85 per cent) in unilateral hydronephrosis. The PI was significantly higher in severe hydronephrosis compared with mild hydronephrosis. In four cases of unilateral multicystic kidney, the PI was always higher on the affected side. Colour Doppler flow mapping and pulsed Doppler evaluation may be helpful in our understanding of renal vascularization in renal pathology and in confirming the diagnosis of renal agenesis. PMID- 8415419 TI - The diagnostic potential of fetal renal biopsy. AB - The feasibility of fetal renal biopsy has been investigated in order to assess the diagnostic value of the histological specimen. Two fetuses with a severe bilateral renal abnormality (multicystic dysplastic kidney, Meckel-Gruber syndrome with polycystic kidney) and one fetus with Down syndrome (no detectable structural anomaly) were sampled. Histological findings in the biopsy specimens of cases 1 and 2 were diagnostic of an early obstructive renal disease. In case 3, the findings were consistent with normal development for gestational age of the kidney. Fetal renal biopsy is technically feasible; histological examination of the samples showed a good correlation with postnatal findings. Further studies of its diagnostic value are required. PMID- 8415420 TI - First-trimester maternal serum Schwangerschafts protein 1 (SP1) in pregnancies associated with chromosomal anomalies. AB - The relationship between first-trimester maternal serum Schwangerschafts protein 1 (SP1) and the karyotype of the pregnancy was examined in 692 women who underwent chorionic villus biopsy at 6-12 weeks. There were 30 pregnancies with abnormal karyotypes, consisting of 14 Down's syndrome (DS), eight trisomy 18, and eight other anomalies, two of which were mosaics. The normal ranges and medians for gestation were defined from the 662 cases in which the karyotype was normal. The median SP1 (0.5 MOM) of the abnormal group was significantly lower than that of the normal group (1.0 MOM). This relationship was maintained for the DS pregnancies (0.4 MOM) and for anomalies other than trisomy 18 (0.43 MOM) but not trisomy 18 (1.1 MOM). It is possible that the use of SP1 as a screening test for chromosome anomalies in the first trimester could have a 43 per cent detection rate for a 5 per cent false-positive rate. PMID- 8415421 TI - A rare inherited euchromatic heteromorphism on chromosome 1. AB - Extra genetic material that is euchromatic is generally regarded to be associated with phenotypic abnormalities. However, recent studies suggest that this is not always the case. Chromosome analysis was performed on amniotic fluid cells from a 37-year-old phenotypically normal patient referred for advanced maternal age. All the cells analysed showed a karyotype of 46,XY,1p+. The 1p+ chromosome had extra genetic material of uncertain origin in chromosome band region 1p21-->31. Chromosome analysis on the father revealed a normal 46,XY male karyotype. The mother's karyotype showed the same 1p+ chromosome. C and Q banding, as well as silver staining studies, in both the mother and the fetus support the interpretation that the extra chromosomal material was euchromatic in nature. This 1p+ chromosome may be characterized as a euchromatic heteromorphism. Euchromatic heteromorphisms not associated with phenotypic abnormalities have been reported for chromosomes 9 and 16. To the best of our knowledge, this is the first report involving this type of cytogenetic anomaly on chromosome number 1 in a phenotypically normal mother and infant. PMID- 8415422 TI - Walker-Warburg syndrome: prenatal ultrasound findings. AB - The prenatal sonographic findings in a case of Walker-Warburg syndrome are described. The patient was not at risk for this condition. Ultrasound examination at 34 weeks' gestation revealed hydrocephaly, Dandy-Walker anomaly, and striking ocular abnormalities. From a review of the literature it appears that while intracranial abnormalities can lead to the diagnosis in cases at risk for this syndrome, ocular abnormalities are rather characteristic for this syndrome and they should be searched for in every case of hydrocephaly or encephalocele. PMID- 8415423 TI - Prenatal diagnosis by in situ hybridization on uncultured amniocytes: reduced sensitivity and potential risk of misdiagnosis in blood-stained samples. AB - Maternal cell contamination was assessed in 18 macroscopically blood-stained amniotic fluid samples from male fetuses. The samples were analysed by double target fluorescent in situ hybridization (ISH) with Y and X chromosome-specific probes. The only sample with an aberrant karyotype (47,XY, + 18) was also analysed by hybridization with a chromosome 18-specific probe. An interpretation of extensive maternal cell contamination was made in two samples, one of which was the sample with trisomy 18. ISH with the chromosome 18-specific probe on this latter sample showed that the sensitivity of the ISH method for chromosome enumeration of uncultured amniotic fluid samples may be reduced in blood-stained samples. It was calculated that by using ISH for chromosome enumeration of the two extensively contaminated samples, a case of trisomy 21 might have been overlooked in both samples, while a case of trisomy 18 might only have been overlooked in one of the samples. It is concluded that ISH should not be used for chromosome enumeration of uncultured amniotic fluid samples that are macroscopically blood-stained without further technical developments. PMID- 8415424 TI - First-trimester prenatal diagnosis of osteogenesis imperfecta type II by DNA analysis and sonography. AB - Osteogenesis imperfecta type II was diagnosed prenatally by analysis of DNA obtained from chorionic villus sampling (CVS) performed at 12 weeks of gestation in a woman who previously had had an affected child. The father had been shown to be mosaic for a mutation in the gene (COL1A2) which encodes the alpha 2(I) chain of type I collagen. An affected fetus was predicted by detection of the mutation in amplified chorionic villus genomic DNA. Ultrasound examination at 13 weeks 4 days demonstrated femoral deformity and virtual absence of calvarial mineralization. In pregnancies at risk for osteogenesis imperfecta type II, sonographic evidence of skeletal abnormalities may be evident by 13 weeks' gestation. PMID- 8415425 TI - Transvaginal sonographic detection of skeletal anomalies in the first and early second trimesters. PMID- 8415426 TI - Emotional reactions in women in late pregnancy (24 weeks or longer) following the ultrasound diagnosis of a severe or lethal fetal malformation. AB - We studied the emotional reactions of 41 women in late pregnancy shortly after they had been informed of the diagnosis of 'severe or lethal fetal malformations' and 3 months after delivery. In addition, situational variables were explored as determinants of grieving. While grief did not diminish during the study period, psychological instability was less pronounced at 3 months after delivery. More grief reactions were evoked by self-reported easily versus self-reported not easily initiated pregnancy, gestational age between 24 and 34 weeks versus beyond 34 weeks, multiparity versus primiparity, and viewing versus not viewing the baby. PMID- 8415427 TI - Attitudes of women of childbearing age towards prenatal diagnosis in southeastern France. AB - The objective of this study was to explore women's attitudes towards prenatal diagnosis of trisomy 21 and to examine some of the factors possibly responsible for these attitudes before implementing in real practice serological screening of pregnant women at risk for trisomy 21. We carried out a telephone survey on a representative sample of women who had recently had a normal livebirth delivery in the Marseille district in 1990. The participation rate was 80 per cent and the average age of the mothers was 28.9 years. Among the 514 women interviewed, 78 per cent stated that they would ask for an amniocentesis for a 1 per cent risk of trisomy 21 at their next pregnancy. When adjusting for confounding factors, the decision to have or not to have an amniocentesis was found to depend not only on the women's attitude towards induced abortion, but also on their understanding of the risk involved and on the social context (knowing a handicapped child, discussion with the father). It also depended on the women's age and on what they knew about amniocentesis from the medical point of view. The risk of miscarriage can influence a woman's choice but this objection was not found to affect the women's decisions significantly in our survey. The data showed the existence of a high potential demand for fetal karyotyping. PMID- 8415428 TI - Prenatal diagnosis of gastrointestinal obstruction: a correlation between prenatal ultrasonic findings and postnatal operative findings. AB - The impact of prenatal sonographic diagnosis of oesophageal and gastrointestinal obstructions has been analysed over a 10-year period. Three groups of patients were evaluated. The first group consisted of 46 newborns with abnormal prenatal sonograms, 41 of which were confirmed to have intestinal obstruction postnatally. The second group consisted of 17 neonates with normal prenatal sonograms who had intestinal obstruction postnatally. The third group included 56 newborns who did not undergo a prenatal sonogram but who had intestinal obstruction confirmed at surgery. Polyhydramnios without the appearance of a stomach on ultrasound was diagnostic of pure oesophageal atresia. Polyhydramnios with intestinal dilation was diagnostic of intestinal obstruction. Although surgery was performed earlier in the infants diagnosed prenatally with ultrasound, mortality was no less than in the group that did not undergo a prenatal sonogram, probably because of the high incidence of associated anomalies. PMID- 8415429 TI - 46,XY/47,XY, + 17p + mosaicism in amniocytes associated with fetal abnormalities despite normal fetal blood karyotype. AB - 46,XY/47,XY, + 17p + mosaicism was found in two primary amniotic fluid cultures (AFCs). Fetal blood karyotype was normal, but ultrasonography revealed Dandy Walker malformation and bilateral choroid plexus cysts. Following termination of pregnancy, fetal examination revealed post-axial polydactyly and neuroblastoma-in situ affecting both adrenals in addition to the cerebellar abnormalities. Mosaicism for the aberrant cell line was confirmed in all fetal tissues sampled and in the placenta. PMID- 8415430 TI - Bilateral pleural effusion at 8.5 weeks' gestation with Down syndrome and Turner syndrome. PMID- 8415431 TI - Prenatal diagnosis of Huntington's disease. PMID- 8415432 TI - [The disease entity as a heuristic principle]. PMID- 8415433 TI - [Persorption of microparticles]. AB - Solid, hard microparticles, such as starch granules, pollen, cellulose particles, fibres and crystals, whose diameters are well into the micrometre range, are incorporated regularly and in considerable numbers from the digestive tract. Motor factors play an important part in the paracellular penetration of the epithelial cell layer. From the subepithelial region the microparticles are transported away via lymph and blood vessels. They can be detected in body fluids using simple methods: only a few minutes after oral administration they can be found in the peripheral blood-stream. We observed their passage into urine, bile, cerebrospinal fluid, the alveolar lumen, the peritoneal cavity, breast milk, and transplacentally into the fetal blood-stream. Since persorbed microparticles can embolise small vessels, this touches on microangiological problems, especially in the region of the CNS. The long-term deposit of embolising microparticles which consist of potential allergens or contaminants, or which are carriers of contaminants, is of immunological and environmental-technical importance. Numerous ready-made foodstuffs contain large quantities of microparticles capable of persorption. PMID- 8415434 TI - [Morphology of bladder cancer with reference to cystectomy specimen]. PMID- 8415435 TI - [Characterization of various types of stomach cancer with differential metastasis in liver, peritoneum and bone]. PMID- 8415436 TI - [Human papillomavirus infections and carcinoembryonic antigen expression in cervix intra-epithelial neoplasia of intermediate grade]. AB - The objective was to investigate a possible correlation between expression of carcino-embryonic antigen (CEA) and human papilloma virus (HPV) infection in moderate cervical intraepithelial neoplasia (CIN II). We examined the correlation of the incidence in different cases and the topographical correspondence. CEA was detected by immunohistochemistry and HPV infection was diagnosed by in situ hybridization (including typing of 6/11, 16/18, 31/33) and well-established histologic criteria. Samples from 32 women were examined. In 29 cases (91%) CEA was expressed, mainly in the superficial layer of the epithelium. Twenty-three tissue samples (72%) showed HPV infection. There was topographical correspondence of the two markers (mainly in superficial cell layers) but no correlation concerning the incidence of HPV infection or particular HPV subtypes. The highest prevalence of HPV was found in dysplasias of the large-cell reserve cell type. PMID- 8415437 TI - [Association and concordance of parameters of Helicobacter pylori infection of gastric antrum mucosa]. PMID- 8415438 TI - [Genetic morphologic fatal syndromes. Neu-Laxova syndrome]. PMID- 8415439 TI - [Genetic morphologic fatal syndromes. Roberts syndrome]. PMID- 8415440 TI - [Alveolar rhabdomyosarcoma with primary manifestation in bone marrow]. PMID- 8415441 TI - [Pleural defect, cause of massive, right-sided hepatic pleural effusion]. PMID- 8415442 TI - [Mycotic panophthalmitis in generalized aspergillosis]. PMID- 8415443 TI - [Food remnant embolism of the lung]. PMID- 8415444 TI - [The history of pathology in Bonn. The difficulties in founding an institute]. PMID- 8415445 TI - [Major hepatic excisions]. PMID- 8415446 TI - [Early diagnosis of silent atherosclerosis and detection of cardiovascular risk factors. A new strategy for the approach of cardiovascular prevention]. AB - The authors present a new strategy for cardiovascular disease prevention based on risk factor detection in occupational medicine and silent atherosclerosis detection in a specialized investigation centre. Employees from several firms in the Paris region were searched, in their working place, for cardiovascular risk factors, including blood cholesterol measurement. The subjects at risk thus selected underwent non-invasive explorations aimed at an early detection of silent atherosclerosis. Extracoronary plaques in the carotid, aorta and femoral arteries were detected by high-resolution ultrasonography, and coronary calcifications by ultrafast CT. The prevalence of arterial lesions and their relationship with risk factors were analysed in a subgroup of 208 untreated male subjects with high blood cholesterol level: 74 percent of these subjects had extracoronary plaques and 65 percent had coronary calcifications. This high prevalence of silent arterial lesions suggested that hypercholesterolaemia, even when moderate, has an early but inconsistent atherogenic effect. Moreover, extracoronary plaques and coronary calcifications were related to risk factors other than blood lipids, and among these factors age was predominant. The simultaneous detection of extracoronary and coronary lesions has demonstrated that extracoronary ultrasonography of several arteries is a good diagnostic test predicting the presence of coronary calcifications in the absence of coronary symptoms. Detection of silent atherosclerosis in subjects at risk therefore is an original and helpful complement to risk factor detection. It should better refine and individualize the diagnosis of risk and evaluate the effects of preventive cardiovascular treatments on atherosclerosis. PMID- 8415447 TI - [Alpha heavy chain disease. Molecular analysis of a new case]. AB - The NH2-terminal structure of serum abnormal protein, as well as the sequence of the corresponding mRNA, were determined in a new case of alpha heavy chain disease. The patient presented with typical clinical features of the disease. Intestinal and mesenteric lymphoplasmic infiltration was monoclonal as assessed by the study of the configuration of heavy and light chain genes. The serum abnormal alpha chains included two molecular species: one starting at the beginning of the hinge region and the other being two amino acids shorter, missing the two first amino acids of the hinge region. The sequence of the mRNA displayed a leader exon, a 93 bp sequence of unknown origin and the second and third constant exons of human alpha 1 chain. These data are discussed in the light of previously reported molecular studies in heavy chain diseases. PMID- 8415448 TI - [From embryogenesis to pathogenesis of effects of diethylstilbestrol on the female genital tract]. AB - From studies on embryogenesis, the authors describe the development, not conditioned by sex steroids, of the upper female genital tract and, in particular, the formation of its cervico-vaginal portion and the shaping of the uterine cervix in about the 16th week of pregnancy. Basing themselves on the physiology of the foeto-placental unit and on the hormonal profile of the foetal ovary, they show that the action of oestrogens on the foetal uterine cervix can be observed only from the 32nd week onward. The action of diethylstilboestrol--a potent oestrogen that is not steroidal and therefore not degraded by foetal metabolism--on the foetal genital tract is explained. An excessive development of the uterine canal caudal part determines structural anomalies of the uterine cervix at birth, the T shape of the uterine cavity and the vaginal diaphragms. The aetiogenesis of clear-cell adenocarcinoma in the cervico-vaginal portion is discussed. PMID- 8415449 TI - [Acute respiratory distress in adults disclosing myopathy caused by acid maltase deficiency]. PMID- 8415450 TI - [Thyroid metastasis of rectal adenocarcinoma]. PMID- 8415451 TI - [Antiphospholipid antibodies associated with cardiac myxoma simulating collagen vascular disease]. PMID- 8415452 TI - [What is still valid in Maranon's law?]. PMID- 8415453 TI - [Pulmonary site of visceral leishmaniasis in HIV infection]. PMID- 8415454 TI - [Shock wave: mechanical effect of electric shock on the atrium]. PMID- 8415455 TI - ["Ciguatera", an uncommon form in La Reunion]. PMID- 8415456 TI - [Hydatid cyst of the pancreas. A case]. PMID- 8415457 TI - [Primary Toxoplasma infection in patients with HIV infection]. PMID- 8415458 TI - [Creutzfeldt-Jakob disease secondary to treatment with growth hormone]. PMID- 8415459 TI - [Erythroblastopenia: chronic idiopathic or associated with chronic lymphoid leukemia. Value of cultures of erythroblastic progenitors and therapeutic strategy]. AB - Fifty cases of chronic pure red cell aplasia in adults (idiopathic in 32 cases, associated with chronic lymphocytic leukaemia in 18) were followed up after studying their erythroblastic precursors in vitro. Analysis of response to immunomodulators confirmed the existence, in patients with idiopathic chronic pure red cell aplasia, of a relationship between in vitro behaviour and obtention of a remission: patients with normal differentiation of erythroblasts consistently responded to treatments, whereas treatments were usually ineffective when the erythroblasts did not differentiate in vitro. When in vitro differentiation was below normal, responses to treatments were varied. In patients with chronic lymphocytic leukaemia, the erythroblastic precursors were normal in 15/18 cases, and response to immunomodulators was observed in 13/18 cases. A second objective of this study was to determine, on a large series, the most adequate immunomodulator treatment. In idiopathic chronic pure red cell aplasia the most efficient therapy was corticosteroids and cyclophosphamide administered separately or together. The antilymphocyte serum gave disappointing results. In pure red cell aplasia associated with leukaemia cyclophosphamide was the most frequently effective drug, with corticosteroids taking second rank; however, infections were frequent. The treatments to be considered when both corticosteroids and cyclophosphamide fail are discussed. PMID- 8415460 TI - [Predictive factors for healing of esophagitis. Prospective study during ranitidine therapy]. AB - This study was conducted to identify factors predicting favourable outcome after ranitidine 300 mg o.d. treatment for oesophagitis. Patients with grade 2 or 3 erosive oesophagitis were included in the study. Each investigator recruited 2 patients with the same grade of lesion in order to minimize bias in assessing symptoms. All patients received the same 6-week treatment: 300 mg ranitidine daily in the form of an effervescent tablet after the dinner. Symptomatology was assessed after 2 and 6 weeks of treatment and endoscopy was performed at the end of the treatment period. A successful outcome was defined as the absence of erosive lesions (grade 0 or 1). A total of 515 patients were recruited and 500 retained for analysis (15 patients did not follow the treatment protocol correctly): 267 grade 2 and 233 grade 3 (mean age: 52.5 years; sex ratio M/F: 1.7). Patients with grade 3 oesophagitis had more severe symptoms; hiatal hernia was also more frequent in this group. Alcohol intake and smoking habits were similar in both groups. After 6 weeks of treatment, successful healing was observed in 87 percent of the grade 2 patients and in 72 percent of the grade 3 patients. Three factors predicting successful outcome were found: grade 2 (odds ratio (OR): 1.50; p = 0.001), symptom duration less than 30 days after treatment onset (OR: 1.50; p = 0.001) and complete relief of symptoms after two weeks of treatment (OR: 1.31; p = 0.03). Disease history and the course of symptoms during treatment with ranitidine could help predict endoscopic outcome of oesophagitis. PMID- 8415461 TI - [Post-traumatic schizophrenia. A case]. AB - We present a case of post-traumatic schizophrenia the interest of which lies in the plurality of aetiological factors and in the signs announcing the disease observed: many mental patients among collaterals and obsessive disorders during childhood. After a brief review of the literature, this case incites us to question the existence of such a thing as post-traumatic schizophrenia and to wonder about the role played by multifactoriality in the determinism of schizophrenia. PMID- 8415462 TI - [Thoracic splanchnectomy under video-thoracoscopy]. AB - The technique of thoracic splanchnicectomy under video thoracoscopic control is described. This little aggressive surgical operation is indicated for very painful forms of pancreatic cancer and for some cases of chronic pancreatitis. It should relieve pain for a longer period than splanchnic nerve injection or radiotherapy. PMID- 8415463 TI - [Microalbuminuria. Signification and value]. AB - The term microalbuminuria means a slightly elevated urinary albumin level. It indicates an abnormal passage of plasma albumin into urine through the glomerular filter. Its prognostic significance was first established in diabetics: in insulin-dependent diabetics microalbuminuria predicts a renal disease, while in non-insulin-dependent diabetics it predicts an excessive cardiovascular mortality. Recently, microalbuminuria has been shown to predict cardiovascular morbidity and mortality in the general population, especially in elderly subjects. Pathogenetic mechanisms common to both atherosclerosis and renal failure may account for the predictive value of microalbuminuria in the above mentioned conditions. PMID- 8415464 TI - [False multiple coin lesions in prostatic cancer under hormone therapy]. PMID- 8415465 TI - [Extensive algodystrophy of the lower limbs in pregnancy]. PMID- 8415466 TI - [Pasteurella multocida, a case of meningitis in a newborn infant]. PMID- 8415467 TI - [Erythroblastopenia in chronic lymphoid leukemia: value of corticoids cyclosporine combination]. PMID- 8415468 TI - [Primary non-Hodgkin's lymphoma of the esophagus in an AIDS infected female]. PMID- 8415469 TI - [Chronic sinusitis caused by Schizophyllum commune in AIDS]. PMID- 8415470 TI - [Intra-parotid nodule caused by Leishmania in a case of apparently cured visceral leishmaniasis]. PMID- 8415471 TI - [Paraneoplastic pemphigus]. PMID- 8415472 TI - [Arthritis in HIV infection. A prospective study of 76 cases in Rwanda]. AB - A 16-month prospective study of adult patients who had been suffering from arthritis for less than one year was conducted in the Internal Medicine department of the hospital of Kigali, Rwanda. Seventy-six patients were included in the study. Out of 72 patients whose sera were tested for human immunodeficiency virus (HIV), 52 were positive, this being 72.2 percent compared with the 20.7 percent figure observed in the urban population of adults in Rwanda. Two main groups arthropathies were individualized: septic arthritis (24 patients including 19 seropositive subjects) and inflammatory rheumatism with criteria of spondyloarthropathies in some of them (34 patients, 32 tested, 28 serum positive). Thus, the finding of acute arthritis in a region with high HIV infection prevalence should suggest a possible HIV infection, especially when the arthritis itself is suggestive of this diagnosis. PMID- 8415473 TI - [Anxiety-related and depressive disorders in women during the premenopausal and menopausal period. Study of the efficacy and acceptability of tianeptine versus maprotiline]. AB - This study was conducted to evaluate the effectiveness and acceptability of Tianeptine (T) versus Maprotiline (M) in the management of anxiodepressive disorders in menopausal and premenopausal women. Anxio-depressive women with a Montgomery Asberg Depression score (MADRS) > 20 and a Hamilton Anxiety score (HARS) > 15 were included in the study. T or M were the only psychotropes taken by the patients during the study. Eighty-three women were enrolled and given, by double blind assignment T 37.5 mg/day or M 75 mg/day (T n = 42; M n = 41). The effectiveness of therapy was assessed on D5, D15, D30 and D60 by the MADRS, HARS, CGI and CHESS scores; acceptability was assessed by the CHESS scale. To exclude placebo responders, the patients were treated with a placebo for 7 days prior to enrollment. A significant improvement in the MADRS score, compared with the previous score, remained until D60 for the T group and M group (p < 0.01). The improvement in the MADRS score was higher (p = 0.025) in the T group than in the M group. At D60, there was a statistically significant difference between T and M in favour of T (p = 0.04). Similar results were obtain with HARS and CGI (item 1). The incidence of side effect was significantly lower in the group treated with T than in the group treated with M. At any time point, there were more patients in the M group (p < 0.001) who complained of side effects than in the T group. The patients were divided into subgroups according to whether or not they were also receiving Hormone Replacement Therapy (HRT). The group taking HRT and T had a more significant reduction in HARS (p = 0.038) and CHESS (p = 0.015) than the group taking T only. T administrated as a single psychotropic agent showed an improvement in the symptoms of anxio-depression which were significantly more important than in the control group with better control of associated complaints. PMID- 8415474 TI - [Caustic rectal stenosis. Trans-anal resection using an EEA stapler]. AB - An original treatment of caustic rectal stenosis due to abuse of analgesic suppositories is presented. The stenosis had been excluded by distal colostomy during 2 years before reconstruction of the circuit was attempted. Anatomically, the stenosis appeared as a complete diaphragm which could be recanalized by an EEA stapler for circular anastomosis, installed in two sites: endoluminal from the colostomy and perineal. After a 2-year follow-up, the anatomical and functional results are satisfactory. PMID- 8415475 TI - [New applications of electricity in clinical practice]. AB - The living matter is sensitive to electric field impulses, and in particular the membrane structure of cells can be transiently modified under calibrated impulses. This, combined with permeabilization, gives access to the cytoplasmic volume. Moreover, fusion of cells by contact or insertion of receptors is made possible. These acquisitions of fundamental research on electricity-living matter interactions has recently led to practical achievements. PMID- 8415476 TI - [Post-traumatic anterior pituitary insufficiency. Value of magnetic resonance imaging]. PMID- 8415477 TI - [Acute respiratory distress syndrome caused by intra-alveolar hemorrhage in leptospirosis]. PMID- 8415478 TI - [Intraspinal calcifications after nucleorthesis: cortivazol too]. PMID- 8415479 TI - [Hashimoto's thyroiditis, adenocarcinoma and malignant lymphoma of the thyroid. A case of triple association]. PMID- 8415480 TI - [Exacerbation of systemic lupus erythematosus following renal puncture-biopsy complicated by hematoma]. PMID- 8415481 TI - [Finger necrosis related to cold agglutinins disclosing Mycoplasma pneumoniae infection]. PMID- 8415482 TI - [Pure cerebellar infarction. 30 cases]. PMID- 8415483 TI - [Cytokines and anti-cytokines. Promising therapeutic application in rheumatic practice]. PMID- 8415484 TI - [Bronchial provocation tests. Comparison of 2 inhalation methods]. AB - This study was performed to compare two methods of aerosols delivery and inhalation in a bronchial provocation test with carbachol : the stocked method (S) where the aerosol is previously stocked in a spirometer bell before inhalation and the dosimeter method (D) where the aerosol is directly inhaled after nebulization. Fourteen subjects (seven normal and seven asthmatic) underwent bronchial challenges with the two techniques. Bronchial sensitivity was calculated on SRaw-dose response curves to carbachol. SGaw-dose response curves were also constructed and the slopes of these curves were used to measure bronchial reactivity. Within subjects sensitivity values were lower with the D method than with the S method (40 +/- 5 micrograms (S E) versus 450 +/- 50 micrograms respectively in asthmatics, 340 +/- 30 micrograms versus 2350 +/- 130 micrograms respectively in healthy subjects). Reactivity values were higher with the dosimeter method (2,7 +/- 0,7 x 10(-5) versus 27 +/- 4 x 10(-5) in healthy and 10 +/- 1 x 10(-5) versus 106 +/- 16 x 10(-5) in asthmatics). The dosimeter method had greater efficacy, was less time consuming and appeared to be a useful method for carrying out standardised non-specific bronchoprovocation test. PMID- 8415485 TI - [Uveo-papillitis associated with paraparesis caused by HTLV-1 virus]. AB - The human T-lymphotropic retrovirus type I (HTLV-1), isolated in 1980, has been shown to be responsible for two distinct systemic diseases: adult T-cell leukemia and HTLV associated myelopathy (HAM). Recently, an ever increasing number of publications have described other disorders associated with HAM. We report a case of uveitis-optic disc neuritis which, to our knowledge, had not been previously described. The fact that both uvea and optic disc were inflamed would suggest that immune-mediated reactions are involved in the pathogenesis of HAM. PMID- 8415486 TI - [Holter monitoring. Value in silent myocardial ischemia]. AB - The inclusion of the digital system in the Holter recording and analyzing systems and the development of micro-computers have multiplied the possibilities and increased the accuracy of this technique. Analysis of the ST segment has resulted in a definition of the silent myocardial ischaemia syndrome. It has also made it possible to evaluate the importance of myocardial ischaemia in daily life and to assess the efficacy of anti-ischaemic treatments. PMID- 8415487 TI - [Successful repair of 2 cases of tracheobronchial tear in operated patients ventilated by Carlen's tube]. PMID- 8415488 TI - [Diffuse interstitial pneumonia, vitiligo and insulin-dependent diabetes in common variable hypogammaglobulinemia of late manifestation]. PMID- 8415489 TI - [Eosinophilic gastroenteritis]. PMID- 8415490 TI - [Endoscopic ablation of gallbladder calculi through the orifice of cholecystostomy]. PMID- 8415491 TI - [Salmonella enteritidis infection with dilatation of the bile ducts]. PMID- 8415492 TI - [Cardiogenic shock caused by occlusive thrombosis of a mitral valve prosthesis treated by fibrinolysis]. PMID- 8415493 TI - [Crohn disease and systemic lupus erythematosus, a rare association]. PMID- 8415494 TI - [Diabetic nephropathy: detection by a strip test for microalbuminuria (Micral Test)]. PMID- 8415495 TI - [Hypertension in pregnancy, Von Recklinghausen disease and pheochromocytoma]. PMID- 8415496 TI - Ten-year follow-up of the Oslo Youth Study Smoking Prevention Program. AB - BACKGROUND: The Oslo Youth Study was designed to demonstrate that health education in schools can result in lowered rates of smoking and alcohol consumption, improved eating patterns, and increased physical activity. The study was conducted from 1979 to 1981 in six schools in Oslo, Norway. This article presents the 10-year follow-up outcomes of the Oslo Youth Study Smoking Prevention Program. METHODS: In the fall of 1989, former Oslo Youth Study participants were contacted through the mail and asked to complete a short questionnaire containing questions regarding smoking behavior. A total of 796 subjects participated (participation rate of 74.0%). RESULTS: Overall, there were no significant effects of the Oslo Youth Study Smoking Prevention Program on the onset of experimental smoking (ever having smoked) or regular smoking (weekly smoking). Significant intervention effects were observed, however, among baseline nonsmoking males; intervention students reported 30% less weekly smoking (35% vs 50%) and 25% greater nonsmoking status (58% vs 44%) in 1989 than did males having attended the reference schools. No treatment effects were observed for females. CONCLUSIONS: The results suggest that the smoking prevention program might have contributed to reduced long-term onset of smoking among participating male subjects. PMID- 8415497 TI - Do drug prevention effects persist into high school? How project ALERT did with ninth graders. AB - BACKGROUND: This article reports follow-up results during grade 9 for a multisite drug prevention program that curbed both marijuana and cigarette use during junior high. Based on the social influence model of prevention, the curriculum sought to motivate young people against drug use and to teach them skills for resisting pro-drug pressures. METHODS: Thirty schools drawn from eight urban, rural, and suburban communities in California and Oregon were randomly assigned to three experimental conditions, two treatment groups and one control. Students in 20 schools received 11 lessons, 8 during grade 7 and 3 in grade 8; in 10 of the treatment schools, older teens assisted an adult teacher in program delivery. Students were pretested prior to the program (grade 7) and post-tested 24 months later (grade 9). RESULTS: Earlier effects on cognitive risk factors (perceived consequences of drug use, normative beliefs, resistance self-efficacy, and expectations of future use) persisted through grade 9 in the teen leader schools; in the condition under which adults taught the lessons without teens, the prior beneficial effects on beliefs largely eroded. All of the earlier effects on actual use disappeared by grade 9, regardless of who taught the lesions. CONCLUSION: Continued reinforcement of earlier lessons may be required to sustain prevention gains through the transition to high school. PMID- 8415498 TI - School-based substance use prevention programs: why do effects decay? PMID- 8415499 TI - The Conference on Comprehensive School Health Education, Washington D.C. and Child Health Day Symposium, New York, 1992. Proceedings. PMID- 8415500 TI - The year 2000 initiative: implications for comprehensive school health. AB - Implementing comprehensive school health programs presents one of our nation's key challenges for the 1990s and beyond. Whether we are talking about Goals 2000 or Healthy People 2000, we will meet the challenge only through the combined efforts of the education and health sectors working as partners at the national, state, and community levels and, perhaps most importantly, at the level of the individual elementary and secondary schools. PMID- 8415501 TI - Schools as healthful environments: prerequisite to comprehensive school health? AB - The development of healthy schools to support and nature the well-being of students, teachers, and staff is proposed as a first step toward the goal of comprehensive health education. A focus on healthy schools incorporates elements of an expanded concept of comprehensive health education that demands careful consideration of the physical, psychological, and social environment of the nation's schools--worksite to 51 million (students, teachers, and staff). The active participation of all stakeholders in environmental assessments, health and safety audits, and restructuring of schools is an essential part of the school reform movement. A healthy worksite concept supports the transformation of the school environment to increase "productivity" by enhancing the ability of teachers, staff, and students to function well. PMID- 8415502 TI - Prevention of heart disease beginning in childhood through comprehensive school health: the Heart Smart Program. PMID- 8415503 TI - Improving the health of U.S. children: the need for early interventions in tobacco use. AB - Despite considerable progress, tobacco use continues to be a major public health problem in the United States, killing more U.S. citizens each year than alcohol, cocaine, crack, heroin, homicide, suicide, car accidents, fires, and AIDS combined. There is cause for particular concern about smoking and other tobacco use by children. Although adolescent smoking declined in the late 1970s and early 1980s, the combination of a now-flat daily smoking prevalence rate among high school seniors, high smoking rates among high school dropouts, and increased use of smokeless tobacco argues for renewed efforts in tobacco use prevention. The greater risk associated with tobacco use at early ages suggests the need for comprehensive interventions (i.e., involving policy change, advertising restrictions, health professionals, and comprehensive school health education) before the prime age for tobacco use initiation (i.e., 12 to 14 years old), for repeated reinforcement of these interventions through adolescence, for innovative ways of reaching underserved--particularly high-risk and minority--youth with effective tobacco use prevention efforts, and for research to determine both the short-term and the long-term effectiveness of these approaches. PMID- 8415504 TI - Childhood fitness: what is happening? What needs to be done? AB - BACKGROUND: There is some controversy as to the fitness levels of U.S. school-age youth. Some experts claim that U.S. youth are fit. Others feel that there has been a decline, despite a recent adult fitness boom. METHODS: The purpose of this article is to examine the research conducted on the cardiovascular endurance and body composition levels of U.S. youth. Research studies noting the activity patterns of youth outside and during physical education are presented. The frequency and duration of physical education requirements are also reviewed. The effects of these patterns and requirements on cardiovascular endurance and body composition are presented. RESULTS: U.S. youth do not engage in physical activity, within our outside physical education, sufficient to develop cardiovascular endurance. Our youth are, therefore, at risk of developing a myriad of diseases associated with sedentary lifestyles. CONCLUSIONS: Exercise is known to have a prophylactic effect on disease, death, and disability. Young people must be instructed and encouraged to be involved in lifetime fitness activities if we are going to control health care costs, reduce disease incidence, and improve the overall quality of life of our citizens. PMID- 8415505 TI - Perspectives on teaching comprehensive school health. AB - This paper seeks to give some perspective to health education teaching as it has developed over a period of time. First, I review the source of our modern comprehensive sequential curriculum design--just to remind us of its roots. Following that, I describe current developments in teaching, highlighting some examples of widely used curricula and sources of other teaching aids for health. Finally, I review some relevant findings from a recent evaluation of comprehensive health education in U.S. public schools to show us where we are today and to help answer the questions: "Is comprehensive school health education worthwhile?" PMID- 8415506 TI - An essential strategy to improve the health and education of Americans. PMID- 8415507 TI - A report card to the nation on adolescents and sexually transmitted diseases. AB - During this 10-min presentation, at least 50 adolescents across the United States will acquire one or more sexually transmitted diseases. Some of these diseases can fortunately be treated and cured; however, other diseases will remain with these young people for the rest of their lives, causing dramatic physical and psychological repercussions. During the brief time allotted to me, I will discuss some of the pertinent statistics related to sexually transmitted diseases among adolescents, review the Health Objectives for the Year 2000, and suggest means for realizing these objectives. PMID- 8415508 TI - Youth tobacco use in the United States--problem, progress, goals, and potential solutions. AB - BACKGROUND: Efforts to control tobacco use and tobacco-related morbidity and mortality in the United States continue to be generally successful. In the quarter century since the publication of the first Surgeon General's Report on Tobacco and Health, adult smoking rates in the United States have been reduced by nearly 34%. Controlling tobacco use among our nation's youth, however, has not been as successful. Although there was considerable success in reducing adolescent tobacco use in the late 1970s and early 1980s, tobacco use among youth has remained essentially stable for the past decade. METHODS: The health and economic burden of tobacco use, current knowledge about youth tobacco use, and youth-related national tobacco reduction goals for the Year 2000 are reviewed. RESULTS: Analysis of the research of the past two decades clearly indicates that there is no "magic bullet" in existence or in sight for the reduction of tobacco use, either among youth or among adults. This does not mean that opportunities for significant advances through, for example, pharmacological therapies or the broad application of media or policy strategies should not continue to be explored, but that for the moment no single approach appears to work best. Rather, a comprehensive approach that applies multiple prevention and cessation strategies simultaneously appears to be most effective in tobacco use control. CONCLUSIONS: Among youth, the combination of tobacco control strategies that may work best includes those that involve the family, primary care physicians, and other health professionals such as nurses and dentists; programs that are carried out in schools and/or through the media; and societal approaches such as access and advertising restrictions and increased taxes. PMID- 8415509 TI - Environmental risks to the health of American children. AB - The major environmental health issue for children today is the extremely high prevalence of unacceptable exposure to lead, especially in inner cities, but occurring throughout the country. It is now generally accepted that lead is toxic to the developing nervous system at levels that were thought only a decade ago to be without effect. Children are more susceptible to the effects of lead than the adults who live in the same environments. Although lead-based paint is no longer used and lead is now removed from gasoline, children will continue to live in housing with the potential for lead poisoning for perhaps another generation. Research into the prevention of exposure and prevention of the consequences of unavoidable exposure is now under way. PMID- 8415510 TI - Youth health report card: asthma. AB - Asthma is a serious, chronic illness among U.S. children. Approximately 6% of all children under 18 years of age in the United States suffer from asthma- prevalence rates in our inner cities are even higher. The impact this disease has on children, their families, and society is profound and easily translatable into loss of time at work and school, a concomitant loss of productivity, and added stress to an already overburdened health care system. Recognizing the significance of asthma as a serious public health problem, Healthy People 2000 has targeted improvements in asthma care as one of its objectives and the National Heart, Lung, and Blood Institute has research and education programs to improve the health of our children with asthma. PMID- 8415511 TI - Communication disorders in children: a challenge for health care. AB - BACKGROUND: Today's children will participate in a society whose economic basis will be information and communication and they must have optimal communication skills to be able to be functional optimally within the information society. Heretofore, in the United States, there has been no systematic attempt to diagnose and care for children with communication disorders. METHODS: It is essential that the child be diagnosed early because there appears to be a critical period for language development. CONCLUSION: If interventions do not occur early, in the area of communication disorders, there is a high probability of significant morbidity because of impaired communication abilities. This morbidity can manifest itself in a social activity and loss of economic opportunity. PMID- 8415512 TI - Youth fitness--problems and solutions. AB - Review of the current data in the area of youth fitness reveals some alarming trends. Children in the United States are fatter, slower, and weaker than their counterparts in other developed nations. In addition, U.S. children seem to be adopting a sedentary lifestyle at earlier ages. Although there is no easy solution to this problem, there are specific recommendations that can ensure improvement in this area. The cornerstone for any meaningful change must involve programs that seek to increase physical activity both in school and at home. Daily, quality physical education in grades K-12 should be mandated in all states. Parents should be educated regarding the critical importance and the multitude of benefits to be derived from their involvement in fitness-related activities with their children. A healthy balance must be established between sedentary activities, e.g., television and video games, and physical activity. All schools should establish fitness testing programs for children and these should be based on health-fitness parameters rather than on athletic performance variables. To ensure improvements in youth fitness across our nation, other interventions are also necessary. These include appropriate involvement of local communities, state and federal governments, the medical health professions, and the media. Specific strategies are available for each group. PMID- 8415513 TI - Summary: school health education. PMID- 8415514 TI - Alcohol and other drug use. PMID- 8415515 TI - A child health report card: 1992. AB - It is because of statistics like the ones included above in the Report Card that the health of our children has become a topic of great concern. These statistics, however, reflect only a small piece of a much larger problem, which includes child poverty, child neglect, child abuse, family disintegration, educational failure, violence, and crime. Indeed the biggest threats to child health have roots in the past and present core of our social and environmental conditions. Improving the health of our children will require innovative and comprehensive approaches that include health education, health services, and family support. The cost of our failure to fund preventive programs in the area of child and family health is significant and mounting. Prenatal care for a pregnant women for 9 months cost about $600; however, medical care for a premature baby for only one day may cost more than four times as much ($2,500). Similar comparisons for the cost of prevention versus treatment are listed in Table 8. It is clear that unless we as a nation place more emphasis on funding preventive medicine, the health of our children will continue to suffer, with grave consequences for the future of our country. PMID- 8415516 TI - [The effect of dietary fiber on proteolysis]. AB - In the presence of food fibers, proteolysis occurs with the formation of sorption complexes. At the initial stages of hydrolysis, the adsorption of both casein and pepsin is observed, the adsorption influencing the initial rate of hydrolysis. The composition of the sorption layer changes during hydrolysis in favor of intermediates. The initial and stationary rates of hydrolysis depend on the protein/fiber ratio. The catalytic activity of the system decreases both with a little amount and with an excess of fibers. The highest rate of hydrolysis is observed at a protein/fiber ratio of 1/2. PMID- 8415517 TI - [Identification of aurofusarin in Fusarium graminearum isolates, causing a syndrome of worsening of egg quality in chickens]. AB - Twenty isolates of Fusarium graminearum have been isolated from infested grain. When laying hens were fed with biomass of most isolates, the quality of eggs deteriorated. It was found that all the isolated produced a yellow-orange metabolite with an antibiotic activity against mycelial fungi and yeasts. The metabolite was identified by physico-chemical methods as the dimeric naphthoquinone aurofusarin. The production of the other mycotoxins zearalenone and deoxynivalenol by the fungal isolates did not evoke the syndrome of egg quality deterioration. PMID- 8415518 TI - [Serum apoproteins and lipoproteins in diabetic children]. AB - Serum levels of total cholesterol, triglycerides, lipoproteins (VLDL, LDL, HDL) and apoproteins (apo-AI, apo-AII, apo-B) were measured in 112 children (70 boys, 42 girls) aged 6 to 13 with type I diabetes mellitus and healthy controls. Poorly controllable diabetes was associated with elevated concentrations of total cholesterol and low-density lipoproteins C and lowered levels of high-density lipoproteins C, apo AI and apo-AII, while differences in triglyceride, very low density lipoproteins, and apo-B were insignificant. Children with a good metabolic control (HbA1 8.8 +/- 1.2%) had a much more favorable lipid profile. The differences may be explained by lipid-reducing effects of diets and insulin therapy. Deficiency of HDL-C, apo-AI and apo-AII in type I diabetes in children indicates the importance of HDL subclasses and of lipolytic enzymes evaluation for an early detection of atherosclerosis risk. PMID- 8415519 TI - [Use of domestic human insulin in patients with type I diabetes mellitus]. AB - Twenty-five patients with type I diabetes mellitus (8 women and 7 men) aged 19 to 54 with a disease duration of 3 to 19 years were treated by Russian semisynthetic short-acting human insulin mono-H. The daily dose varied from 24 to 44 U. The drug therapeutic efficacy was assessed from the terms of attainment of carbohydrate metabolism compensation, glycosylated hemoglobin level, and changes in daily insulin requirement. Clinical safety of the drug was assessed from changes in antiinsulin antibody level, incidence and severity of hypoglycemia, absence of lipodystrophy and other adverse reactions. The drug was found effective for the treatment of patients with type I diabetes. Its hypoglycemic effect was similar to that of actrapid, a porcine insulin. Fractionated insulin therapy resulted in the majority of patients in a reduction of glycosylated hemoglobin level, of daily requirement in insulin, and of antiinsulin antibody titer. PMID- 8415520 TI - [Improvement of diet therapy in children with insulin-dependent diabetes mellitus]. AB - The authors emphasize that determination of the glycemic post-alimentary effects for various foodstuffs (white bread, mashed potatoes, vermicelli, milk gruels: semolina, buckwheat, rice, porridge, millet) represents a new approach to dietetic therapy of diabetes mellitus. Low glycemic indexes of rice gruel and vermicelli, compatible to those of buckwheat gruel, recommend these food stuffs for diabetics' rations. Determination of glycemic indexes will help classify the food by its glycemic effect and thus help optimize dietetic therapy of diabetes mellitus. PMID- 8415521 TI - [Glurenorm in the treatment of patients with non-insulin-dependent diabetes mellitus with diseases of the liver and bile ducts]. AB - Glurenorm, a IInd generation sulfanylurea preparation, was used for a year as a sugar-reducing drug in 20 patients with non-insulin-dependent diabetes mellitus and concomitant diseases of the liver (cirrhosis, chronic hepatitis, n = 5) and biliferous duct (cholelithiasis, a state following cholecystectomy, chronic cholecystitis, n = 15). A year follow-up has not shown deterioration of liver function as indicated by results of liver tests (AST, ALT, acid phosphatase, gamma-glutamyltranspeptidase, bilirubin, cholesterol, triglycerides). The hypoglycemic effect of the drug proved to be inferior to that of sulfanylurea derivatives, but the absence of side effects permit higher doses of glurenorm (up to 4-6 tablets daily) as against other oral sugar-reducing drugs. PMID- 8415522 TI - [Pathogenetic rationale for the treatment of obesity]. AB - On the basis of experience gained in prolonged follow-up of 1095 obese patients, studies of their hormonal homeostasis using computer-aided cluster analysis the author comes to a conclusion that division of obesity cases into constitutional exogenous and hypothalamic is conditional. The hypothalamic syndrome develops in 100% of obesity cases. A course of pathogenetic therapy paralleled by a subcaloric diet and exercise therapy is suggested. Bitemporal inductothermy proved to be effective. The new complex for obesity treatment includes low caloric diets combined with pathogenetic drug therapy and physiotherapeutic exposure of the mid-brain. PMID- 8415523 TI - [Effect of squid liver fat and krill meat on blood serum atherogenicity in patients with type I diabetes mellitus]. AB - Study of the effects of squid liver fat and krill meat on blood serum atherogenic characteristics in patients with type I diabetes has shown that these foodstuffs had a certain atherogenic effect, for their consumption was associated with a marked reduction of blood serum atherogenicity. The mechanism of this effect is still to be researched. PMID- 8415524 TI - [Cytogenetic changes in peripheral blood lymphocytes in people with impaired thyroid status]. AB - Increased counts of cells with chromosome aberrations were revealed in peripheral blood lymphocytes of subjects with thyroid dysfunction: 5.5 +/- 0.6% in hypothyrosis, 9.0 +/- 1.8% in hyperthyroidism, and 2.5 +/- 0.1% in controls (20 donors). The aberrations are mainly presented by paired and solitary fragments, rarely by chromatid translocations; in two patients with hyperthyroidism dicentrics were detected. Distribution of aberration types varied. In hypothyrosis chromosome aberrations were two times more incident than chromatid ones. In hyperthyroidism chromosome and chromatid aberrations were approximately equally incident. These results may be used to explain the causes of unfavorable outcomes of pregnancy in patients with thyroid diseases. PMID- 8415525 TI - [Clinical aspects of screening and diagnosis of congenital hypothyroidism in neonates in Moscow]. AB - Screening of the newborns helped diagnose congenital hypothyrosis in infants of Moscow and define its incidence as 1 per 5000 newborns. Seventeen newborns (20%) with congenital hypothyrosis were detected among 65 ones with positive tests of more than 5 microU/ml. Clinical manifestations of this disease may be detected by specialists within the first weeks of life. Blood TTH level higher than 100 microU/ml is an evidence of congenital hypothyrosis, though one case with a lower TTH level in a newborn was detected. Levothyroxine therapy in a daily dose 50 micrograms is the optimal for the majority of such patients. For infants with transitory hypothyrosis with blood TTH levels from 40 to 80 micrograms/ml short levothyroxine therapy courses in daily doses 25-50 micrograms for 2-3 weeks may be recommended to preserve the intellectual potential of the child. All the children with blood TTH levels higher than 20 microU/ml should be regularly examined by endocrinologists and neuropathologists starting from the first days of life. PMID- 8415526 TI - [Diet therapy of diabetes mellitus (lecture)]. PMID- 8415527 TI - [Principles of training patients with diffuse toxic goiter]. PMID- 8415528 TI - [Age-related differences in macromolecule biosynthesis reactions in rat pituitary cells to catecholamine, serotonin and acetylcholine]. AB - In experiments using primary monolayer cultures norepinephrine, isoproterenol, and serotonin (10(-6)-10(-5) M) inhibited the rate of total protein biosynthesis more effectively in pituitary cells of pubertal and sexually mature rats as compared to neonates and prepubertal animals. Acetylcholine (10(-5)M) reduced the rate of total protein synthesis in pituitary cells of pubertal rats but did not change this parameter of functional activity in pituitary cells of neonatal animals. Norepinephrine and isoproterenol inhibited the rate of DNA synthesis in pituitary cells of newborn rats whereas serotonin and acetylcholine were of no avail in this respect. These results permit us suggest an important role of neurohumoral agents in regulation of rat pituitary cells growth and functional activity during the postnatal development. PMID- 8415529 TI - [Experimental study of adrenergic modulation of insulin sensitivity]. AB - Relationships between adrenergic exposure (stress, administration of adrenomimetics and adrenoblockers) and rat sensitivity to exogenic insulin were studied. The participation of beta-adrenoceptors in regulation of insulin sensitivity of the body was revealed. Drugs characterized by beta-adrenomimetic activity (adrenalin, novodrin, partusisten) enhanced insulin sensitivity upon prolonged (for 24 h) administration. One-hour immobilization stress also enhanced insulin sensitivity. The authors discuss beta-adrenoceptor desensitization phenomenon as a possible mechanism of the before events. Obsidan (a nonselective beta-adrenoblocker) also augmented insulin sensitivity. A single administration of partusisten (an adrenomimetic with a predominant affinity to beta 2 adrenoceptors) reduced insulin sensitivity. Augmentation of beta-adrenergic processes at the expense of beta-adrenoblockers (aminasin, haloperidol, butyroxan) effects also reduced insulin sensitivity. An inverse correlation between the capacity of adrenergic exposures to influence glycogen content in striated muscles and their effects on insulin coma latency (insulin sensitivity market) was detected. This relationship confirms the role of beta-adrenoceptors in insulin sensitivity regulation. PMID- 8415530 TI - [Development of hypothalamic regulation of gonadotropic function of the pituitary in hypoinsulinemic rats]. AB - Effects of temporary hypoinsulinemia induced by streptozotocin in neonatal age on the development of the mechanism of hypothalamic regulation of hypophyseal gonadotropic function in rats were studied. With this aim the authors analyzed the status and functional capacity of feedback system mediating hypothalamic regulation of gonadotropin secretion in female and male rats at the age when this system starts functioning in health. Streptozotocin injection in the neonatal age induced a reduction of estradiol concentration and of estradiol and progesterone stimulated preovulatory peak of luteinizing hormone (LH) in 28-day-old females and a reduction of LH concentration in 28-day-old castrated males. Reduced estradiol and LH peak were observed in adult females injected streptozotocin during the first week of life and reduced LH level in adult males. These results permit a hypothesis that streptozotocin administration to rats in the neonatal age disturbed the mechanism of hypophyseal gonadotropic function regulation in mature rats. PMID- 8415531 TI - [Role of norepinephrine in the regulation of thyroid gland functional activity in rabbits]. AB - By collecting in situ the blood flowing from the thyroid, the authors have demonstrated that noradrenaline injected in a dose 1 microgram/kg into rabbit thyroid artery enhanced triiodothyronine secretion without changing thyroxin secretion. This process was inhibited by prazosin, an alpha-adrenoblocker, in a dose 1 microgram/kg, injected 5 min before noradrenaline, but not by propranolol, a beta-adrenoblocker. Phenylephrine, an alpha-adrenergic agonist, in a dose 1 microgram/kg, injected into the thyroid artery, influenced thyroid hormone secretion similarly as noradrenaline. The T3/T4.100 ratio taken as an indicator of T4 thyroid conversion increased under the effect of noradrenaline injection after prazosin. Thus, in our experiment noradrenaline activated thyroid function via stimulation of alpha-adrenoceptor system. Hence, catecholamines are capable of directly influencing thyroid functional activity. PMID- 8415532 TI - [A hypothetical model of the action of steroid hormones]. AB - The authors suggest the existence of at least two different binding sites of hormone receptor complex in a nucleus. These nuclear binding sites have various functions: metabolic and specific acceptors. This hypothesis may explain some peculiarities of steroidal action. PMID- 8415533 TI - [Correlations between specific binding of labeled adrenoligands of the brain and plasma testosterone in mice]. AB - Correlations between specific binding of labeled adrenoligands in the frontal cortex and hypothalamus and peripheral blood plasma testosterone were studied in BALB/cLac, CBA/Lac, and S57BL/6J male mice. A negative correlation was detected between blood testosterone and the extent of specific binding of 3H-clonidine, a 1.6 nM alpha 2-adrenoceptor agonist, in the hypothalamus. No correlations were revealed between specific binding of 3H-clonidine in the frontal cortex and binding of the rest radioligands (0.4 nM alpha 1-adrenoceptor antagonist 3H prazosin and 1.6 nM beta-adrenoceptor antagonist 3H-dihydroalprenolol) in both brain segments and blood testosterone level. Further experiments in six mouse strains (A/He, BALB/cLac, CBA/Lac, C57BL/6J, DD, and YT) revealed a reliable negative genotypic correlation between hypothalamic maximal specific binding of 3H-clonidine and testosterone level. The detected feedback may be an important regulatory mechanism of the hypothalamo-hypophyseo-gonadal system in male mice. PMID- 8415534 TI - [Effect of passive lymphocyte transfer on the course of autoimmune thyroiditis in guinea pigs]. AB - Effects of adoptive transfer of T lymphocytes from normal animals (intact and in vitro pretreated with tactivin) on the expression of experimental autoimmune thyroiditis in guinea pigs were studied. Thyroiditis development was characterized by immunocompetent cell imbalance, formation of antithyroid antibodies, development of diffuse and focal lymphoid-plasmacytic infiltration in thyroid tissues. A single injection of intact T lymphocytes normalized the immunity status and significantly (by three times) reduced the lymphoid infiltration area but did not augment thyroxin secretion in animals with thyroiditis. Immunotherapy of the condition with tactivin-activated lymphocytes was found more effective, for it enhanced serum thyroxin secretion, normalized the immunity status, and reduced lymphoid-plasmacytic infiltration of the thyroid. PMID- 8415535 TI - [Thyroglobulin]. PMID- 8415536 TI - [Hormonally-inactive pituitary adenomas]. PMID- 8415537 TI - [Microcirculatory hemostasis in children with insulin-dependent diabetes mellitus]. AB - Study of microcirculatory hemostasis in children with insulin-dependent diabetes mellitus helped the authors to make new conclusions on metabolic changes underlying blood coagulation cellular factors, namely, the red cell and platelet levels. Fifty children aged 7 to 15, suffering from type I condition, were examined. Ten of these were examined during the compensation stage, 23 during decompensation without ketosis, and 17 with ketoacidosis. Twelve age-matched healthy children were controls. Noticeable disorders in the microcirculatory hemostasis system were revealed in diabetic children, these changes manifesting by changed functional activities of red cells and platelets. Reduced share of platelet deaggregation, increased red cell aggregation time, reduced thromboelastogram chronometric constant and reduction of its structural parameters may be considered as the early signs of these changes. The risk of microcirculatory hemostasis disturbances augments with the disease duration and progress. PMID- 8415538 TI - Presence of an endogenous superoxide dismutase activity in three rodent malaria species. AB - Superoxide dismutase (SOD) was investigated in three species of rodent malaria (Plasmodium berghei, P. yoelii and P. vinckei). The isoelectric points (pI) of isozymes found in purified parasites were identical. SOD activities detected by isoelectrofocusing at pl 5.0, 5.6, and 6.4 were cyanide-sensitive and could be considered as having been adopted by the parasites from the host red blood cell. The three rodent malaria parasites also contained a cyanide-resistant, hydrogen peroxide-sensitive SOD activity not found in the host red blood cell. It is therefore concluded that the three rodent malaria parasites possess an endogenous SOD. Two bands of endogenous SOD were found at pl 6.2 and 6.8 for the three species, and one additional band was detected at pl 5.7 for P. berghei and P. vinckei. This first report in rodent Plasmodium of a cyanide-resistant, hydrogen peroxide-sensitive SOD suggests that these parasites may be capable of at least partly resisting activated oxygen species using an endogenous SOD. PMID- 8415539 TI - Entamoeba histolytica: changes in the zymodeme of cloned nonpathogenic trophozoites cultured under different conditions. AB - In this paper we report the occurrence of changes in the migration of certain isoenzymes of a cloned strain (MAV-CINVESTAV) of Entamoeba histolytica. This strain was isolated from an asymptomatic carrier and showed an initial nonpathogenic zymodeme I. The transfer of polyxenic trophozoites from Robinson's medium (7% serum) to Jones' medium (30% serum) provoked changes in isoenzyme expression, resulting in the conversion of zymodeme I to a zymodeme that was similar to the XVII zymodeme except for two slow-running bands and a single fast running band that were detected for hexokinase (HK). This XVII-like zymodeme reverted to zymodeme I when trophozoites were cultured under monoxenic conditions in TY1-S-33 medium (10% serum). When we cultured cloned strain MAV-CINVESTAV under axenic conditions in TY1-S-33 medium, trophozoites expressed a pathogenic zymodeme with two fast-running HK bands and a beta-phosphoglucomutase band. In addition, phagocytosis and the ability of the trophozoites to destroy cell culture monolayers were expressed only in trophozoites cultured under axenic conditions. The axenization procedure required the presence of live Fusobacterium symbiosum and is also described herein. PMID- 8415540 TI - Ultrastructural hepatic alterations in hamsters and jirds after experimental infection with the liver fluke Opisthorchis viverrini. AB - Changes in the hepatocytes of male hamsters (Mesocricetus auratus) and jirds (Meriones unguiculatus) at 220 days after experimental infection with the liver fluke Opisthorchis viverrini were studied by light and electron microscopy. The hepatocytes of the control group were characterized by an intracellular compartmentation. A globular nucleus was located centrally. The main features of the perinuclear zone were the cisternae of the rough endoplasmic reticulum (RER) and interjacent mitochondria, lysosomes, and peroxisomes. The peripheral cell region was dominated by glycogen fields and scattered lipid droplets, which were surrounded by anastomosing tubules of the smooth endoplasmic reticulum (SER). An immense proliferation of the SER was striking in the hepatocytes of animals infected with O. viverrini. Coincidentally, the intracellular compartmentation disappeared. Glycogen rosettes, RER, lysosomes, and lipid droplets were distributed irregularly all over the cell, the latter being observed more frequently than in control animals. The nuclei showed lobe-like protrusions and were enlarged. The mitochondria were often dumbbell-shaped and showed pathologic degenerations up to lysis. Our results resemble those of numerous investigations concerning hepatocellular alterations caused by N-nitroso compounds. Therefore, these observations suggest a synergistic effect for trematode infection and N nitroso compounds in the pathogenesis of opisthorchiasis. The cellular alterations observed in the hepatocytes of Opisthorchis-infected animals together with the accumulation of intermediate filaments seen in the adjacent bile-duct epithelia and in the epithelium of the gall-bladder seem to indicate a disturbance of the cell metabolism and might be related to a neoplastic transformation. PMID- 8415542 TI - Philophthalmus species, probably P. palpebrarum, in Israel: description of the eye fluke from experimental infection. AB - Following a recent incident of human philophthalmosis in Israel, the intramolluscan larval trematode fauna in snails randomly collected from the suspected water source was checked. Of the snails examined, only Melanopsis praemorsa shed cercariae, including a Philophthalmus cercaria. To identify the philophthalmid species involved, chicks were experimentally infected with metacercariae of the trematode, subsequently yielding mature trematodes resembling those of P. palpebrarum. The majority of trematodes obtained, whether from one-worm infections or from multiple-worm infections resulting in a single trematode in one of the eyes, were relatively small and showed only immature eggs in their uteri. This finding suggests that the existing descriptions of two species of Philophthalmus purportedly harbouring eggs with non-oculate miracidia, namely P. palpebrarum and P. nyrocae, are actually based on immature specimens from one-worm infections that precluded cross-fertilisation. If this be true, then all species of the genus Philophthalmus produce eggs that, when mature, contain oculate miracidia. The species encountered in Israel is thus most likely P. palpebrarum. PMID- 8415541 TI - Biochemical and immunological studies on soluble antigens of Entamoeba histolytica. AB - The soluble antigens of Entamoeba histolytica trophozoites were analysed in detail by biochemical and immunochemical methods. The antigen was highly complex and heterogeneous as revealed by Sephacryl S-300 column chromatography, which showed four distinct fractions. The molecular mass of fractions FI, FII, FIII and FIV was 660, 170, 65 and 13 kDa, respectively. Protein was the major constituent in crude soluble antigen (CSA) and fractions FI and FII (67%, 80% and 90%, respectively). Polysaccharide was predominant in the FIII fraction (59%). Antigenic activity observed after different physico-chemical treatments revealed that CSA and FI antigens were predominantly glycoprotein in nature. However, the antigenicity of FIII antigen was greatly reduced after sodium meta-periodate treatment, whereas no alteration in reactivity was discerned after trypsin treatment. Sodium dodecyl sulphate-polyacryl-amide gel electrophoresis (SDS-PAGE) analysis demonstrated nearly 28 Coomassie blue bands for CSA and 20, 16, 15 and 3 polypeptide bands for the FI, FII, FIII and FIV fractions, respectively. The molecular mass of the polypeptides of these bands ranged from 210 to 20 kDa. Antigenic activity was observed in CSA and in the first three fractions, both in counter immunoelectrophoresis (CIEP) and in enzyme-linked immunosorbent assay (ELISA). However, the highest antigenic activity was noted in fraction FI. Major immunoreactive polypeptides of CSA and FI antigens against whole trophozoite antibody were observed in the 10- to 170-kDa regions. However, major differences in the immunoreactivity of the two antigens were noted at 116 and 14 kDa for FI antigen and at 84, 30 and 20 kDa for CSA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415543 TI - Serum-free cultivation of Plasmodium falciparum gametocytes in vitro. AB - Several components were tested for their ability to replace human serum in cultures of Plasmodium falciparum set up for the development of gametocytes. Besides a serum-free medium, A*I*M*V (Gibco BRL), RPMI medium supplemented with commercially available serum substitutes was used to culture gametocytes. The following substances served as serum replacements: Basal Medium Supplement (Biochrom), Ultroser G (Gibco BRL) and Nutridoma-SR (Boehringer Mannheim). All serum-free additives supported some parasite growth, but only in RPMI supplemented with Nutridoma-SR were morphologically mature gametocytes obtained. The asexual forms developed almost as well as in RPMI with human serum added. PMID- 8415544 TI - Isoenzyme variation within the genus Cryptosporidium. AB - Soluble extracts of the oocysts of Cryptosporidium parvum had demonstrable, but low, activities of malate dehydrogenase (MDH, EC. 1.1.1.37), carboxylesterase (ES, EC 3.1.1.1) and lactate dehydrogenase (LDH, EC. 1.1.1.27) following thin layer starch-gel electrophoresis. Much higher activities of glucose phosphate isomerase (GPI, EC. 5.3.1.9) and phosphoglucomutase (PGM, EC. 2.7.5.1) were found, and zymograms of these two enzymes were used to characterise isolates of C. parvum from human, bovine, ovine and cervine sources, C. muris from the brown rat and C. baileyi from young turkeys. PGM and GPI zymograms clearly distinguished between C. parvum, C. muris and C. baileyi. The five isolates of C. parvum showed the same electrophoretic mobility for GPI, whereas the PGM mobility of the single human isolate of C. parvum examined was clearly different from that of the other isolates. This is the first report of the use of isoenzymes to distinguish between species and isolates of Cryptosporidium. PMID- 8415545 TI - Comparative effects of anthelmintics on c-MDH from Molinema dessetae (Nematoda: Filarioidea) and from a mammal. AB - Malate dehydrogenase (MDH) (E.C.1.1.1.37) activity was detected in the filaria Molinema dessetae at a level similar to those found in other filariae. In M. dessetae, the cytoplasmic form (c-MDH) predominated and the study was performed on partially purified fractions. The pH optimum for oxaloacetate reduction was 6.1, with maximal activity at 7811 nmol min-1 mg protein-1, but high concentrations of oxaloacetate inhibited MDH activity. The Km value for oxaloacetate was determined as 22 microM for M. dessetae c-MDH and 33 microM for mammalian c-MDH. Anthelmintic drugs were compared as potential inhibitors of filarial and mammalian c-MDH. Among the compounds evaluated, amocarzine showed a specific inhibitory effect on filarial c-MDH through only at high concentrations. Suramin alone showed an inhibitory effect at low concentrations (Ki = 1.15 microM) but without selective action towards filarial c-MDH. The suramin type of inhibition was found to be competitive. Suramin probably acts on both enzymes in the same manner. Nevertheless, M. dessetae c-MDH is proposed as a suitable enzyme assay model to screen MDH inhibitors as potential filaricides. PMID- 8415546 TI - Kinetics and pattern of organelle exocytosis during Toxoplasma gondii/host-cell interaction. AB - The fate of Toxoplasma gondii dense-granule (GRA2, GRA3), rhoptry (ROP1), and surface (SAG1) proteins was followed by immunofluorescence assay (IFA) and immunoelectron microscopy at different stages after infection. Dense-granule exocytosis occurred in the apical area of the tachyzoite within minutes of invasion. Several exocytic events were found simultaneously in the same organism, both by serial sectioning and by freeze-fracture studies. Dense-granule contents were first found as a dense material trapped between parasite and vacuole membranes before either the vacuolar network or the vacuole membrane could be immunolabeled with specific antibodies. The vacuolar network was strongly labeled with dense-granule antibodies but not with the SAG1-specific probe, which suggests that the network is not enriched in membrane proteins. In addition to strongly labeling the vacuole membrane, GRA3 antibodies also labeled strands extending from the parasitophorous vacuoles into the host-cell cytoplasm. PMID- 8415547 TI - Electron immunogold labeling of regulatory peptide immunoreactivity in the nervous system of Moniezia expansa (Cestoda: Cyclophyllidea). AB - An electron immunogold-labeling technique was used in conjunction with a post embedding procedure to demonstrate for the first time the ultrastructural distribution of the parasitic platyhelminth neuropeptide, neuropeptide F (NPF), in the nervous system of the cestode Moniezia expansa. Two axon types, distinguished by their populations of different-sized electron-dense vesicles, were identified. Immunogold labeling demonstrated an apparent homogeneity of PP, FMRFamide and NPF (M. expansa) antigenic sites throughout the larger dense-cored vesicles within the central nervous system. Triple labeling clearly demonstrated the co-localisation of immunoreactivities (IR) for NPF, PP and FMRFamide within the same dense-cored vesicles. The presence of NPF-IR within the vesicles occupying the perikaryon of the neuronal cell body indicated that the peptides had undergone post-translational C-terminal amidation prior to entering the axon. Antigen pre-absorption experiments using NPF prevented labeling with either PP or FMRFamide antisera, and the failure of these antisera to block NPF-IR supports the view that some, if not all, of the PP/FMRFamide-IR is due to NPF-like peptides. PMID- 8415548 TI - Physico-chemical conditions necessary for the in vitro excystment of Zygocotyle lunata (Trematoda:Paramphistomatidae). AB - Various physico-chemical factors associated with the in vitro excystment of metacercariae of Zygocotyle lunata were studied. Metacercariae could excyst with or without the presence of trypsin or bile salts, although the rate of excystation was slower in the absence of both. Crude ox-bile extract promoted faster excystment than did more refined sodium taurocholate. Pretreatment of cysts with pepsin also resulted in faster excystment rates. Optimal excystation occurred at 40 degrees C and between pH 5.5 and 7.5. The broad pH and temperature ranges and the non-specific enzyme/bile salt requirements for excystation were correlated with the wide range of vertebrate hosts used by this trematode. PMID- 8415549 TI - Genetic processes within an epidemic of sleeping sickness in Uganda. AB - Reproductive processes within the current Ugandan epidemic of sleeping sickness are investigated. Genotype frequencies derived from isoenzyme patterns in 44 stocks of Trypanosoma brucei s.l. collected in 1988 from Tororo, south-east Uganda are analysed by single and multiple loci methods. In the single locus method, the hypothesis of random mating is tested by agreement with Hardy Weinberg equilibrium. The multiple loci method uses a contingency table approach to detect non-random associations between pairs of loci; this equates to the detection of disequilibrium. The results do not support the concept of a randomly mating population of T. brucei within the current epidemic. Results from the epidemic data set are discussed in relation to the broader problem of genetic exchange in Trypanozoon. PMID- 8415550 TI - The common frog (Rana temporaria) as a potential paratenic and intermediate host for Angiostrongylus vasorum. AB - Common frogs (Rana temporaria) were exposed either to third-stage larvae (L3) or to first-stage larvae (L1) of Angiostrongylus vasorum. Following exposure to L3, viable larvae could be detected in the frogs for at least 2 weeks. Following exposure to L1, the frogs developed viable L3 in their tissues within 30 days. In a test of the infectivity of these larvae, one fox was fed frogs previously infected with L3 and another fox was fed frogs previously infected with L1. On autopsy it was found that adult A. vasorum populations had been established in both foxes. Thus, it could be concluded that frogs can act not only as paratenic hosts but also as intermediate hosts for A. vasorum. PMID- 8415551 TI - Immune defense and eosinophilia in congenitally IgE-deficient SJA/9 mice infected with Angiostrongylus costaricensis. AB - The roles of IgE in protective immunity and eosinophilia in Angiostrongylus costaricensis infection were examined by comparing IgE-deficient SJA/9 mice and IgE-producing SJL/J mice. In primary infection, mean total IgE levels increased to a maximum of 390 ng/ml, which was more than 10 times greater than the 29 ng/ml measured preinfection in SJL/J mice but less than the 10 ng/ml found in SJA/9 mice throughout the experiment. Immune defense as determined by recovery of adult worms and eosinophilia were similar in SJL/J and SJA/9 mice. Protective immunity was induced by infection with A. costaricensis followed by treatment with levamisole for 4-6 days postinfection. After the challenge infection, the numbers of adult worms and eosinophils in SJA/9 mice were not significantly different from those in SJL/J mice. Anti-A. costaricensis IgE antibody was not detected in either strain of mice during the experiment. These results indicate that A. costaricensis infection induced the production of IgE not specific for parasite antigens in IgE-producing mice. Potentiated nonspecific IgE played no significant role in immune defense and eosinophilia. PMID- 8415552 TI - Identification of truncated forms of lipophosphoglycan in mutant cloned lines of Leishmania major that are deficient in mature lipophosphoglycan. PMID- 8415553 TI - Detection of chitin in spores of Myxobolus muelleri and M. subepithelialis (Myxosporea, Myxozoa). PMID- 8415554 TI - Effects of mebendazole on Angiostrongylus costaricensis in mice, with special reference to the timing of treatment. AB - Mebendazole was given to mice infected with Angiostrongylus costaricensis at a single dose of 5 mg/kg at 6, 11, 16 or 21 days post-infection (p.i.) and in five successive doses at 5 mg/kg daily at 6, 11 or 16 days p.i. The effects were comparatively assessed by examining various parameters in host mice and worms. As a whole, the effects of mebendazole were caused more conspicuously by five successive treatments than by a single treatment. In both treatment modalities, the effects were more remarkable in earlier treatments, and nearly complete effects were caused by five successive treatments before 15 days p.i. These results suggest that the inhibition of egg formation and/or oviposition will inhibit the pathological changes caused in the disease by A. costaricensis, especially before the onset of the changes. PMID- 8415555 TI - Prevalence of agglutinating anti-Leishmania antibodies in two multi-thousand Bengoli communities. AB - Control of endemic visceral leishmaniasis (VL) in large communities requires a feasible epidemiological indicator capable of monitoring on-going transmission rather than mere exposure to the parasite. Following confirmation of the desired level of reliability for laboratory diagnosis of VL, the direct agglutination test (DAT) was employed to estimate VL sero-prevalence in the endemic upazilas (subdistricts) of Trishal and Shahjadpur within Mymensingh and Sirajganj districts of Bangladesh. DAT antigen production was duly increased to allow coverage of a study population of 17826 inhabitants, 9619 of whom resided in Trishal, 7328 in Shahjadpur and 879 in Teknaf (Cox's Bazar), a known Leishmania free district in Bangladesh. Despite large-scale production in batches of 1120 4000 ml (each sufficient for 1176-6400 screening doses), all DAT antigen batches performed as required in quality control tests for sensitivity, specificity and stability. It was convenient for both collection and testing to take the required samples of whole blood by finger prick. A cross-sectional survey revealed VL point prevalences of 4.40% in Trishal and 6.75% in Shahjadpur, compared with an extremely low rate of 0.34% in non-endemic Teknaf. In both endemic upazilas (Trishal and Shahjadpur) VL was more prevalent (2.56-4.5%) in persons up to 20 years of age than in those 21 years of age and older (1.84-2.25%). Of 918 subjects recorded as seropositives, 539 were VL-asymptomatic and 379 were VL symptomatic with various degrees of suspicion. Diagnosis of VL was established in 125 symptomatic seropositives subjects, either by demonstrating the presence of Leishmania amastigotes (29), or by positive DAT results combined with presentation of typical VL signs (96).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415556 TI - Molecular biology studies of tubercidin resistance in Trypanosoma cruzi. AB - Trypanosomatids are incapable of de novo purine synthesis; purines are obtained through the scavenging of exogenous nucleosides. To advance our understanding of purine utilization, we mutagenized a Trypanosoma cruzi stock and selected for resistance to high levels of tubercidin (7-deazaadenosine, TUB), a purine analog. The TUB-resistant stocks were > 100 times more resistant to TUB than was the parental stock. TUB and uridine transport in the TUB-resistant stocks decreased by 50%-90%, whereas thymidine and adenosine transport were unaffected. These data imply that TUB-resistant stocks have defects in the pathways involved in the transport of TUB and uridine but not in the thymidine and adenosine transport pathways. Karyotype analyses using specific probes showed that the deletion of a 950-kb chromosome-size DNA occurred in both of the TUB-resistant stocks. These data suggest that genes involved in nucleoside transport are located in this DNA region. This study will facilitate the identification and characterization of the specific genes involved in nucleoside transport and aid in the elucidation and development of new chemotherapeutics for Chagas' disease. PMID- 8415557 TI - In vitro encystment and experimental infections of Blastocystis hominis. AB - Cultures of Blastocystis hominis were induced to encyst using three encystation media: (a) an encystation medium (EM) comprising yeast extract in buffered saline containing 50% horse serum, (b) an encystation medium (CEM) comprising EM conditioned with bacterial soluble products and (c) an encystation medium (TEM) containing 0.5% trypticase in EM. Two isolates of B. hominis were studied, an axenized isolate C and a non-axenized isolate MS. In EM, isolate C did not encyst, whereas 6.1% of isolate MS had encysted by day 1. However, in CEM and TEM, 17.4% and 25.7% of isolate C, respectively, had encysted by day 5. In all three media, isolate MS encysted more readily than isolate C, with as much as 91.7% of the former encysting in TEM. As viewed by phase-contrast microscopy, cyst-like stages appeared highly refractile. Direct stool examination of juvenile Wistar rats infected with 10,000 cyst-like stages of both C and MS isolates showed Blastocystis at day 2 post-infection. At autopsy on day 7, large numbers of Blastocystis were seen in the cecum, with smaller numbers being observed in the large intestine. In contrast, rats fed with various inocula of the vacuolar stages of isolates C and MS did not become infected, indicating the importance of the encysted stages in the transmission of the parasite. PMID- 8415558 TI - Differentiation of Trichinella isolates by polymerase chain reaction. AB - Oligonucleotide primers were synthesized for the polymerase-chain-reaction amplification of target DNA from two sequences of Trichinella spiralis. Six strains belonging to T. spiralis, T. nativa, T. britovi, T. pseudospiralis, and T. nelsoni were tested. Amplification products were obtained with T. spiralis, T. britovi, and T. nelsoni DNA from a 53-kDa antigen cDNA sequence and with T. spiralis and T. nelsoni DNA from a 1.6-kb repetitive DNA sequence. Differences in the length of the amplification products obtained from the repetitive sequence would enable a differentiation between T. spiralis and T. nelsoni, suggesting that the 1.6 kb repetitive DNA sequence would not be specific for T. spiralis. No amplification was detected for T. nativa or T. pseudospiralis DNA from the two sequences and for T. britovi DNA from the 1.6-kb repetitive DNA sequence. PMID- 8415559 TI - Theileria sergenti infection in the Bo-RBC-SCID mouse model. AB - We have previously developed the Bo-RBC-SCID mouse model for Theileria sergenti infection. In the present study, this model was further examined to delineate the models of parasite infection. The Bo-RBC-SCID mice were prepared by periodically transfusing uninfected bovine erythrocytes (Bo-RBCs) into splenectomized SCID mice via the intraperitoneal (i.p.) route. The mice, separated into three groups, were inoculated i.p., intravenously (i.v.), or subcutaneously (s.c.) with T. sergenti-infected Bo-RBCs. Examination of samples of peripheral blood demonstrated that the parasite infected mice inoculated via any one of the three routes. The mice inoculated i.v., however, developed parasitemia earlier than those inoculated i.p. or s.c. When Bo-RBC-SCID mice prepared without splenectomy were infected with T. sergenti, a high-level parasitemia appeared only once. After that, not only the level of parasitemia but also the number of Bo-RBCs in the peripheral blood rapidly decreased despite the continuation of Bo-RBC transfusions. The results suggest that T. sergenti proliferates primarily in the circulating blood in Bo-RBC-SCID mice and that in response to the parasites growth, the spleen may play an important role in the removal of both parasitized and unparasitized Bo-RBCs from the blood circulation. PMID- 8415560 TI - Fatty acid composition of Echinostoma trivolvis (Trematoda) rediae and adults and of the digestive gland-gonad complex of Helisoma trivolvis (Gastropoda) infected with the intramolluscan stages of this echinostome. AB - Gas-liquid chromatographic studies were done to determine the fatty acid composition of the digestive gland-gonad (DGG) complex of Helisoma trivolvis snails infected with the intramolluscan stages of Echinostoma trivolvis, of rediae freed from the DGG, of uninfected DGG, and of 41-day-old adult worms grown in golden hamsters. The DGG of infected snails showed significantly higher levels of stearic acid (18:0), hexatrienoic acid (16:3n-4), and docosahexanoic acid (22:6n-3) than that of uninfected snails. However, the DGG of uninfected snails showed significantly higher levels of 20:2 non-methylene-interrupted diene (NMID) and adrenic acid (22:4n-6) than that of infected snails. The profiles of other fatty acids were remarkably similar in both infected and uninfected snails. Adult worms showed significantly higher amounts of numerous saturated fatty acids and dienes as compared with the rediae. However, the rediae showed significantly higher amounts of certain monoenes and trienes as compared with the adults. Fatty acid differences between rediae and adults probably reflect differences in either the available lipid pools in the immediate host sites or the metabolic activity of each stage of this echinostome. PMID- 8415561 TI - Molecular modeling: a tool for predicting anthelmintic activity in vivo. AB - The structural and electronic features of the broad-spectrum benzimidazole anthelmintic mebendazole [MBZ, methyl 5-(benzoyl)-benzimidazole-2-carbamate] have been determined using a combination of quantum mechanics, molecular graphics, and molecular modeling techniques. Using conformational analyses and quantum mechanics, we found that the three-dimensional structure and electronic features of MBZ were consistent with those previously reported for highly active broad spectrum benzimidazole anthelmintics and that in vivo drug efficacy against Hymenolepis diminuta depends upon the orientation of the benzoyl group at position 5 on the heterocyclic ring system, the magnitude of the molecular dipole moment, and the percentage of polar surface area. The chemotherapeutic actions of MBZ on H. diminuta in vivo were accompanied by marked changes in worm weight and chemical composition. Tapeworms recovered from rats that had received a therapeutically effective dose of MBZ 24 h earlier were significantly smaller and contained much less glycogen (as a percentage of the wet weight) than worms from untreated controls. In MBZ-treated worms, protein concentrations rose at a rate sufficient to offset the decline in glycogen concentration. Glycogen/protein ratios in MBZ-treated worms were considerably lower than the corresponding control values. Differences in the absolute amounts of glycogen between control and drug-treated worms were even more profound. Administration of a curative dose of MBZ to the rat host produced in H. diminuta another change, the onset of which coincided with the gross alterations in worm weight and chemical composition.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415562 TI - Variant proteins associated with ionophore resistance in sporozoites of Eimeria tenella (Coccidia). AB - Protein samples prepared from sporozoites of two ionophore-sensitive strains (WIS and Penn St) and three resistant strains (FS139, FS459, and FS462) of Eimeria tenella were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), native polyacrylamide gel electrophoresis (native PAGE), or two-dimensional electrophoresis with native PAGE and SDS-PAGE. Variant proteins that might be associated with ionophore resistance were observed in resistant field strains on native PAGE. When two-dimensional electrophoresis was conducted, four over-expressed peptides with approximate molecular weights of 97, 71, 70, and 50 kDa were observed in the FS139 field strain in comparison with the WIS laboratory strain. PMID- 8415563 TI - Coexistence of heterogeneous populations of Toxoplasma gondii parasites within parasitophorous vacuoles of murine macrophages as revealed by a bradyzoite specific monoclonal antibody. AB - The expression of bradyzoite-specific antigens (Bsa) of Toxoplasma gondii was studied in murine bone-marrow-derived macrophages that had previously been infected with tachyzoites. Growth conditions that allowed only restricted replication of Toxoplasma gondii resulted in heterogeneous populations; (1) Bsa positive and Bsa-negative parasites could be observed within one parasitophorous vacuole (heterogeneous vacuole community), and (2) homogeneous Bsa-positive and homogeneous Bsa-negative vacuole communities coexisted within one macrophage host cell. These observations suggest that stage conversion does not seem to be a synchronous event for a vacuole community. PMID- 8415564 TI - Coprological study of the Dicrocoelium dendriticum (Digenea) egg elimination by cattle in highland areas in Leon Province, northwest Spain. AB - Dicrocoelium dendriticum egg output in cattle from five locations of the Porma river basin (Leon Province, Northwest Spain) was studied at monthly intervals between March 1986 and March 1987. We found D. dendriticum eggs in 37.64% of the 1251 samples examined, and the number of eggs per gram (epg) ranged from 10 to 1000 (average, 41.65 +/- 2.73). The main egg-elimination period, for prevalence, was autumn-winter. The average epg values were quite similar during all months except March, when the maximum was detected. The infection prevalence increased in cattle aged up to 2 years and then a slight, gradual decrease was observed with increasing host age. The average epg values ranged from 40.08 +/- 3.24 to 48.33 +/- 12.42 in all age groups except the oldest animals, for which the figures were lower. The rate of egg elimination was higher in animals that came from locations situated at a greater altitude. PMID- 8415565 TI - Influence of Leishmania infection on blood-meal digestion in the sandflies Phlebotomus papatasi and P. langeroni. AB - The presence of amastigote-initiated infections of Leishmania major parasites caused a significant suppression in alkaline protease, trypsin and aminopeptidase activity during the first 30 h after ingestion of the infected bloodmeal in Phlebotomus papatasi, the natural vector of L. major. Protease levels were significantly higher in infected flies after 72 h than in the control group, where digestion had ceased. Evidence for the suppression of protease activity in infected P. langeroni, a sympatric but un-natural vector of L. major, was less clear; there was no difference in alkaline protease activity between control and infected groups in the first 24 h. However, protease, trypsin and aminopeptidase activities were elevated after 72 h in infected P. langeroni, indicating a delay in the time to the end of digestion and passage of the bloodmeal. The potential advantages for parasite development in suppressing protease activity and extending the period of bloodmeal digestion are discussed. PMID- 8415566 TI - Schizodeme and zymodeme analysis of trypanosomes of the subgenus Schizotrypanum from the bat. AB - Two stocks and nine clones of trypanosomes of the subgenus Schizotrypanum were analyzed based on their electrophoretic pattern of EcoR1 digestion products of kinetoplast DNA (k-DNA) minicircles (schizodeme) and isoenzymes (zymodeme). The trypanosome stocks were isolated from Phyllostomus hastatus bats collected in different locations in Minas Gerais State, Brazil. The k-DNA pattern showed major variations between stocks and microvariations among the clones. Furthermore, in one of several cloning attempts, two different populations could be isolated. Among the six enzymes studied, glucose phosphate isomerase presented different patterns for stocks and clones. No similarity was found among bat trypanosomes and Trypanosoma (Schizotrypanum) cruzi standard zymodemes. This shows that bat trypanosomes are distinct from T. (S.) cruzi. PMID- 8415567 TI - Immunogenicity and antigenic reactivity of a carrier-free synthetic peptide complex derived from a 70-kDa Plasmodium falciparum exoantigen. AB - A 70-kDa Plasmodium falciparum exoantigen was purified from supernatant fluids of continuous in vitro P. falciparum cultures using sequential cation-exchange and high performance liquid chromatographic procedures. The purified protein was then digested with chymotrypsin and amino-acid sequences were determined for the resulting fragments. Four peptides (termed C2, C3, C5, and C10) were subsequently selected for synthesis based on their predictability for antigenic sites. The peptides were effectively used as a synthetic immunogen (SPf70) when they had been copolymerized with glutaraldehyde in the absence of a carrier. When given with Freund's adjuvant, the synthetic peptide complex was found to be highly immunogenic in rabbits. Serologic reactivity to the peptide complex and peptides C2 and C5 was uniformly high, followed by the responses to peptides C3 and C10. Peptide antigenicity was also assessed with human anti-P. falciparum sera from malaria-endemic regions of Uganda and Venezuela. Enzyme-linked immunosorbent assay (ELISA) data showed that anti-P. falciparum antibodies were specific for and reactive to the peptides. The specificity of the rabbit anti-SPf70 antibodies for P. falciparum antigen was shown by immunoprecipitation of metabolically labeled proteins and by immunoblotting. Herein we describe the peptide sequences of a 70-kDa P. falciparum exoantigen (Pf70) that, when synthesized and constructed as a copolymer (SPf70), are capable of inducing the formation of antibodies that are reactive with the native malarial protein. The high immunogenicity and antigenic reactivity of SPf70 indicate the potential use of this synthetic peptide polymer as an immunogen and a diagnostic reagent. PMID- 8415568 TI - Compatibility of one Brazilian and two Venezuelan strains of Schistosoma mansoni with various strains of Biomphalaria glabrata. AB - For evaluation of the degree of genetic heterogeneity in parasite and snail strains, the compatibility between Biomphalaria glabrata and Schistosoma mansoni from different geographical areas was studied. Venezuelan snails from Barbula, Manuare (areas of schistosome transmission) and Tinaquillo (non-transmission area) and Brazilian BH snails were exposed to infection with miracidia of both the Venezuelan YT and SM strains and the Brazilian BH strain of S. mansoni. Snail parasite compatibility was evaluated by quantifying the number of snails shedding cercariae during a period of 35 to 60 days post-infection. The best compatibility appeared to be between the Brazilian parasites and the Brazilian or Venezuelan snails. In contrast, the Brazilian snails appeared to be resistant to infection by the Venezuelan parasites tested. Paradoxically, the compatibility between the sympatric pair of Venezuelan parasites and Venezuelan snails appeared to be lower in comparison with the allopatric association of the Brazilian parasite and the Venezuelan snails. The results suggest an important degree of heterogeneity in the snail and parasite isolates studied and yield biological markers for both organisms. PMID- 8415569 TI - Climate and cuticular hydrocarbon variation in Rhipicephalus sanguineus ticks (Acari: Ixodidae). AB - An association between the climate and variation of the cuticular hydrocarbon pattern was found for several populations of Rhipicephalus sanguineus ticks. Extreme parameters of climate (absolute minimal and maximal temperatures) correlated positively with six compounds (mainly methylalkanes) detected in cuticular hydrocarbon mixtures. In the light of work conducted on cuticular transpiration in arthropods and biochemical phase changes in cuticle lipids, it is suggested that the adaptation of ticks to potential colonization areas is accomplished by detectable changes in cuticular hydrocarbon relationships. These tick strains can be biochemically determined and their adaptation potential, analyzed. PMID- 8415570 TI - Dose- and time-dependent functional and structural damage to the colon mucosa by Entamoeba histolytica trophozoite lysates. AB - Analysis of the initial interaction between Entamoeba histolytica trophozoites and the large intestine is impossible in humans and very difficult in experimental animals. To circumvent this obstacle we treated the luminal side of full-thickness rabbit colon segments mounted in Ussing-type chambers with trophozoite lysates of the E. histolytica HM1 virulent strain. Exposure to lysates for up to 90 min produced dose- and time-dependent effects on the colon, consisting of (a) increased decay rates for potential difference, short-circuit current, and transmural resistance and (b) mucosal damage ranging from vacuolation at the bases and shortening of epithelial cells to the loss of intercellular junctions, destruction of microvilli, and necrosis of interglandular epithelial zones. This acute model of intestinal amebiasis is sensitive, fast, and reliable. PMID- 8415571 TI - Chitin: a cell-surface component of Phytomonas francai. AB - The occurrence of chitin as a structural component of the surface of the phytopathogenic protozoan Phytomonas francai was demonstrated by paper and gas liquid chromatographic analysis of the products of enzymatic and chemical hydrolysis of alkali-resistant polysaccharides, lectin binding, glycosidase digestion, and infrared spectra. Chitin was characterized by its insolubility in hot alkali and chromatographic immobility as well as by the release of glucosamine on hydrolysis with strong acid and of N-acetylglucosamine (GlcNAc) on hydrolysis with chitinase. The presence of chitin was also shown directly by binding of wheat-germ agglutinin (WGA), which recognizes GlcNAc units, to the parasite surface. Fluorescein-labeled WGA binding was completely abolished by treatment with chitinase. This effect was specific since it could be prevented by incubating the enzyme with chitin before treatment of the phytomonads. These findings indicate that chitin is an exposed cell-surface polysaccharide in Phytomonas francai. The data were confirmed by the infrared spectrum of an alkali insoluble residue, which showed a pattern typical of chitin. PMID- 8415572 TI - Experimental transmission of murine malaria by the oral route. AB - A total of 116 young male CD1 mice were orally inoculated with mouse blood; half of the animals received 0.2 ml of uninfected blood and the others were given 0.2 ml of Plasmodium berghei yoelii-infected blood in six experiments performed at different times. Almost 30% of the experimental mice acquired malaria as demonstrated by the observation of parasites in their blood. In no case were parasites found in the blood of control mice. Rodent malaria parasites may be transmitted to CD1 mice by the ingestion of mouse blood parasitized by P. b. yoelii. As far as we know, this study represents the first demonstration of oral transmission of murine malaria. Oral transmission studies in this mouse Plasmodium model may produce very important information on the biology of the malaria parasites. PMID- 8415573 TI - Topological and stereochemical restrictions in beta-sandwich protein structures. AB - Chain topology in beta-structured protein domains and handedness associated with it are discussed. Previously, other workers have shown that by considering just two restrictions--structures that are left-handed and/or have loops that cross can be disregarded--the number of topologies associated with such structures is expected to be severely limited. By way of example, we determine the number of topologies compatible with a six-stranded antiparallel beta-sandwich. Without restriction on the type of strand-strand connection allowed but with elimination of symmetry related structures 360 topologies are possible. If connections between parallel strands are disqualified the number is reduced, 10-fold, to 36. The figure is cut to 24 when structures with loop crossings are eliminated. Handedness in these structures is examined in detail and from this a rationale for the observed predominance of right-handed forms of beta-structures is presented. The 24 structures can be considered as a set of right- and left-handed pairs of 12 topologies. All but two of these pairs can be assigned hands on the basis of existing rules. Six of the structures are found to occur in the Brookhaven Protein Databank and all are right-handed. This study provides a basis for protein design projects which might, for example, attempt the synthesis of unobserved protein topologies. Of the 24 structures in the final set eight are examples of the classic Greek key fold. Thus, the predominance of this motif among all-beta proteins can be attributed in part to these topological constraints. The possible physicochemical origins of the structural selection rules and additional factors which might contribute to the particular favourability of certain structures are also explored. PMID- 8415574 TI - The structure of a designed peptidomimetic inhibitor complex of alpha-thrombin. AB - Thrombin displays remarkable specificity, effecting the removal of fibrinopeptides A and B of fibrinogen through the selective cleavage of two Arg Gly bonds between the 181 Arg/Lys-Xaa bonds in fibrinogen. Significant advances have been made in recent years towards understanding the origin of the specificity of cleavage of the Arg16-Gly17 bond of the A alpha-chain of human fibrinogen. We have previously proposed a model for the bound structure of fibrinopeptide A7-16 (FPA), based upon NMR data, computer-assisted molecular modeling and the synthesis and study of peptidomimetic substrates and inhibitors of thrombin. We now report the structure of the ternary complex of an FPA mimetic (FPAM), hirugen and thrombin at 2.5 A resolution (R-factor = 0.138) and specificity data for the inhibition of thrombin and related trypsin-like proteinases by FPAM. The crystallographic structures of FPA and its chloromethyl ketone derivative bound to thrombin were determined. Although there are differences between these structures in the above modeled FPA structure and that of the crystal structure of FPAM bound to thrombin, the phi, psi angles in the critical region of P1-P2-P3 in all of the structures are similar to those of bovine pancreatic trypsin inhibitor (BPTI) in the BPTI-trypsin complex and D-Phe Pro-Arg (PPACK) in the PPACK-thrombin structure. A comparison between these and an NMR-derived structure is carried out and discussed. PMID- 8415575 TI - Theoretical examination of the mechanism of aldose-ketose isomerization. AB - Two mechanisms for an aldose-ketose isomerization have been examined using high level ab initio and semiempirical molecular orbital methods. The proton transfer pathway via an enediol intermediate is shown to be favored in the absence of a metal ion, while the hydride transfer pathway becomes favored in the presence of a metal ion. Our calculations explain why the proton transfer pathway is operative in most aldose-ketose isomerization reactions. These calculations also provide further support for the previously proposed metal ion-mediated hydride transfer mechanism for xylose isomerase. PMID- 8415576 TI - Identification and classification of protein fold families. AB - We have developed a method for identifying fold families in the protein structure data bank. Pairwise sequence alignments are first performed to extract families of homologous proteins having 35% or more sequence identity. Representatives are selected with the best resolution and R-factor to give a nonhomologous data set. Subsequent structure comparisons between all members of this set detect homologous folds with low sequence identity but highly conserved structures. By softening the requirement on structural similarity, families of analogous proteins are obtained that have related folds but more diverse structures. Representatives are selected to give a non-analogous data set. Starting with 1410 chains from the Brookhaven Data Bank, we generate a set of 150 nonhomologous folds and a set of 112 non-analogous folds. Analysis of sequence and structure conservation within the larger families shows the globins to be the most highly conserved family and the TIM barrels the most weakly conserved. PMID- 8415577 TI - An evaluation of the performance of an automated procedure for comparative modelling of protein tertiary structure. AB - A 3-D model of a protein can be constructed from its amino acid sequence and the 3-D structures of one or more homologues by annealing three sets of fragments: the structurally conserved regions, structurally variable regions and the side chains. The method encoded in the computer program COMPOSER was assessed by generating 3-D models of eight proteins whose crystal structures are already known and for which 3-D structures of homologues are available. In the structurally conserved regions, differences between modelled and X-ray structures are smaller than the differences between the X-ray structures of the modelled protein and the homologues used to build the model. When several homologues are used, the contributions of the known structures are weighted, preferably by the square of sequence similarity; this is especially important when the similarities of the homologues to the modelled structure differ greatly. The 'collar' extension approach, in which a similar region of different length in a homologue is used to extend the framework, can result in a more accurate model. If known homologues comprise more than one related group of proteins and they are both distantly related to the unknown, then alignment of the sequence to be modelled with each group of homologues facilitates identification of structurally conserved regions of the unknown and leads to an improved model. Models have root mean square differences (r.m.s.d.s) with the structures defined by X-ray analysis of between 0.73 and 1.56 A for all C alpha atoms, for seven the eight models. For the model of mucor pepsin, where the closest homologue has 33% sequence identity and 20% of the residues are in structurally variable regions, the r.m.s.d. for the framework region is 1.71 A and the r.m.s.d. for all C alpha atoms is 3.47 A. PMID- 8415578 TI - Introduction of disulfide bonds into Bacillus subtilis neutral protease. AB - The effects of engineered disulfide bonds on autodigestion and thermostability of Bacillus subtilis neutral protease (NP-sub) were studied using site-directed mutagenesis. After modelling studies two locations that might be capable of forming disulfide bonds, both near previously determined autodigestion sites in NP-sub, were selected for the introduction of cysteines. Analysis of mutant enzymes showed that disulfide bonds were indeed formed in vivo, and that the mutant enzymes were fully active. The introduced disulfides did not alter the autodigestion pattern of the NP-sub. All mutant NP-subs exhibited decreased thermostability, which, by using reducing agents, was shown to be caused by the introduction of the cysteines and not by the formation of the disulfides. Mutants containing one cysteine exhibited intermolecular disulfide formation at elevated temperatures, which, however, was shown not to be the cause of the decreased thermostability. Combining the present data with literature data, it would seem that the introduction of disulfide bridges is unsuitable for the stabilization of proteases. Possible explanations for this phenomenon are discussed. PMID- 8415579 TI - Making tissue-type plasminogen activator more fibrin specific. AB - The fibrin specificity of tissue-type plasminogen activator can be increased by mutagenesis within at least four sites in the protease domain. These sites include residue I276, the new N-terminus formed by conversion to a two-chain structure, residues on either side of the active site cleft, KHRR 296-299 or DDD 364-366, a charged surface involved in fibrin interactions, which includes residues H432, R434, D460, R462 and a loop structure, PQANL 466-470, near the fibrin-binding patch. Variants with mutations at any of these sites have low fibrinogen-stimulated activity, whereas fibrin-stimulated activity is at least normal. Kinetic analysis reveals that mutations at these positions reduce the kcat in the presence of fibrinogen, but leave the molecules with normal kinetic constants in the presence of fibrin. A significant exception is found at positions 296-299, where the presence of fibrin manifests significant increases in both kcat and Km. Combinations of mutations at these sites appear to be additive with respect to fibrin specificity. PMID- 8415581 TI - George Orwell's '1984' revisited; in public bureaucracy, failure is success. PMID- 8415580 TI - Errors in the information deposited in gene data banks. PMID- 8415582 TI - Diabetic neuropathy--the presence and future of a common but silent disorder. PMID- 8415583 TI - Long survival with giant cell myocarditis. AB - Several aspects of giant cell myocarditis remain controversial, including the natural history of the disease and the nature of the giant cells. We have observed three patients who had long survival with chronic active giant cell myocarditis. The first patient was a 59-yr-old female who had a 10-yr history of complete heart block which was found at autopsy to have been caused by giant cell myocarditis. The second patient is a 36-yr-old female who received a heart transplant 5 yr after a biopsy proven episode of active myocarditis, and examination of the explanted heart revealed giant cell myocarditis. The third patient was a 41-yr-old male who received a heart transplant 2 yr after developing progressive heart failure, and the explanted heart had giant cell myocarditis. On immunohistochemical study of the three hearts, the giant cells stained with the macrophage markers lysozyme and KP-1 (CD-68). Staining of the same cells with desmin and actin was focally positive in a punctate pattern, correlating with the ultrastructural presence of myofibrils within giant cell phagolysosomes. The associated lymphocytic infiltrate stained primarily for the T cell markers CD-3, CD-45RO, and CD-43 whereas only a few of the lymphocytes stained with the B-cell marker CD-20. The long histories of cardiac dysfunction in the three patients show that giant cell myocarditis may have a protracted course. The morphologic studies show that the giant cells are of histiocytic origin but can contain phagocytosed components of myocytes, observations that may account for the controversy surrounding the nature of the giant cells in giant cell myocarditis. PMID- 8415584 TI - DNA histogram typing in retinoblastomas and neuroblastomas. AB - The nuclear DNA content characterization was carried out by means of both DNA index and DNA histogram type assessments in a series of 21 retinoblastomas, 11 neuroblastomas, 1 ganglioneuroblastoma, and 4 medulloblastomas. These measurements were performed by means of the cytophotometric digital cell image analyses of Feulgen-stained nuclei. The results indicate that as far as nuclear DNA content is concerned, retinoblastomas seem to be very different from neuroblastomas. In fact, in terms of DNA index, retinoblastomas are significantly more aneuploid than neuroblastomas. The DNA histogram type shows that the high level of aneuploidy found in retinoblastomas corresponds to genotypically polymorphic tumors, and this could reflect a serious degeneration of the genomic material in retinoblastomas. This type of degeneration seems to be much less frequent in neuroblastomas, which basically seem to be either diploid or hypertriploid. PMID- 8415585 TI - Pancreatic lipase is a useful phenotypic marker of intrahepatic large and septal bile ducts, peribiliary glands, and their malignant counterparts. AB - Expression of pancreatic lipase in normal, proliferating, and carcinomatous epithelia of the intrahepatic biliary tree was examined by immunohistochemistry in 82 normal livers, 35 hepatolithiatic livers, 11 cholangiocarcinomas (CCs) associated with hepatolithiasis, 34 CCs, and four combined hepatocellular cholangiocellular carcinomas. The intrahepatic biliary tree was anatomically divided into large ducts, septal ducts, interlobular ducts, bile ductules, and peribiliary glands. In hepatolithiasis, large ducts, septal ducts, and peribiliary glands showed marked proliferation and dysplasia. In normal livers and hepatolithiasis, expression of pancreatic lipase was found in large ducts in 91% and 94%, in septal ducts in 95% and 94%, and in peribiliary glands in 93% and 94%, respectively. Interlobular ducts, bile ductules, and hepatocytes were negative for pancreatic lipase. The immunoreactivity of pancreatic lipase was coarse granular, and was regularly present in the supranuclear and to a lesser degree paranuclear cytoplasm of the epithelial cells. All cases of CCs with hepatolithiasis, which arised from large ducts, expressed pancreatic lipase. In CCs, pancreatic lipase was expressed in the perinuclear cytoplasm of cancer cells in 67% in the hilar type and in 24% in the peripheral type (P < 0.02). The combined hepatocellular-cholangiocellular carcinomas failed to express pancreatic lipase in both elements. These data suggest that large ducts, septal ducts, and peribiliary glands contain pancreatic lipase in normal and proliferative conditions, and that CCs probably arising from these ductal elements continue to express pancreatic lipase. Thus, pancreatic lipase could be a phenotypic marker of large ducts, septal ducts and peribiliary glands as well as their malignant counterparts. PMID- 8415586 TI - Brain lymphomas of immunocompetent and immunocompromised patients: study of the association with Epstein-Barr virus. AB - The Epstein-Barr virus (EBV) has been described in association with lymphomas of the central nervous system. To test if the association between EBV and central nervous system lymphomas was limited to patients with immunosuppression or whether the association also held for patients who were immunocompetent and to determine the true prevalence of any association, we studied 37 formalin-fixed, paraffin-embedded cases of central nervous system lymphomas occurring in immunocompromised and immunocompetent patients. EBV DNA-RNA in situ hybridization was performed using a 30-base biotinylated antisense oligonucleotide complementary to the EBER1 gene of EBV. Immunohistochemistry was also performed, using a monoclonal antibody to the latent membrane protein of EBV. The most common histologic subtypes of lymphoma were high grade immunoblastic (57%), diffuse large cell (22%), and small, noncleaved, non-Burkitt's (11%). Eighty-six percent of all tumors in our series were of B-cell lineage, as confirmed by CD20 expression, but only 37% of the primary tumors showed restricted CD20 expression. EBV RNA was identified in all or virtually all of the malignant lymphoid cells in 11 of the 37 cases (30%), including 10 primary brain lymphomas and one metastasis from systemic disease. Latent membrane protein expression was identified in 64% of the EBER1-positive cases. All 10 patients who had a history of immunosuppression had tumors that expressed EBV RNA. We confirm the strong association of EBV with brain lymphomas occurring in immunocompromised patients, whether due to AIDS or to the immunosuppression associated with organ transplantation. Our findings also demonstrate a low rate of EBV-positivity in immunocompetent patients. PMID- 8415587 TI - Low prevalence of human papillomavirus types 16 and 18 in cervical adenocarcinoma in situ, invasive adenocarcinoma, and glandular dysplasia by polymerase chain reaction. AB - Using the polymerase chain reaction on paraffin blocks for human papillomavirus (HPV) types 16 and 18, a positive result was seen in three of 20 cases of invasive adenocarcinoma, 15 of 36 cases of adenocarcinoma in-situ, and one of five cases of glandular dysplasia of the uterine cervix. Types 16 and 18 were found with equal frequency. In adenocarcinoma in situ there was no age difference between HPV-positive and or HPV-negative cases. Concomitant squamous dysplasia was slightly more frequent in HPV-positive adenocarcinoma in-situ. HPV positivity rates are lower than most of those previously reported. Possible reasons for this are discussed. PMID- 8415588 TI - Concurrent mesothelioma and adenocarcinoma of the lung in a patient with asbestosis. AB - We report the apparently unique simultaneous development of a mesothelioma and an adenocarcinoma of the lung in a patient with asbestosis. Pathologists should be aware that very rarely these two malignancies may occur concurrently, an event with potential prognostic, therapeutic, and medical-legal implications for the patient and his family. Although occupational asbestos exposure is well recognized as a risk factor in the development of both mesothelioma and lung carcinoma, this case report emphasizes the rarity of the synchronous occurrence of these tumors in asbestos exposed individuals suggesting that the mechanism by which asbestos fibers induce lung carcinoma is different from that by which they induce mesothelioma. PMID- 8415589 TI - Mutations in the p53 gene in human astrocytomas: detection by single-strand conformation polymorphism analysis and direct DNA sequencing. AB - The p53 gene was examined in a series of formalin-fixed paraffin-embedded astrocytic neoplasms of various types by polymerase chain reaction (PCR), single strand conformation polymorphism analysis (SSCP), and direct sequencing of amplified DNA. PCR primers were designed to amplify three DNA fragments encompassing exons 5, 7, and 8 with splice sites, including all four mutational "hot spots" within this gene. SSCP was performed in a polyacrylamide gel containing 10% glycerol. Two mutations were found among the 20 high and intermediate grade adult astrocytomas studied by this sensitive screening technique and confirmed by sequencing of the PCR product. (1) An anaplastic astrocytoma disclosed a T-A transversion in Codon 246 giving rise to a methionine to lysine amino acid substitution. (2) A giant cell glioblastoma disclosed a G to A transition in Codon 285 resulting in a glutamic acid to lysine substitution. Both mutations were associated with loss of the normal allele. Twenty-three DNA fragments that disclosed no mutation by SSCP analysis were confirmed to be negative by direct sequencing of amplified DNA. No mutations were detected in a series of eight juvenile cerebellar astrocytomas, a biologically distinct form of low-grade astrocytoma. Mutations of the p53 gene may play an important pathogenetic role in a subset of human astrocytomas. PMID- 8415590 TI - pS2 immunostaining of colorectal carcinoma. AB - This study was aimed at assessing the significance of pS2 immunostaining in colorectal adenocarcinoma and adjacent tissue. Paraffin sections of 63 surgically resected colorectal adenocarcinomas were stained with a pS2 specific monoclonal antibody using the avidin-biotin complex immunoperoxidase technique. Several tumor sections also included adjacent nonneoplastic mucosa. Six tubular adenomas and five hyperplasic polyps found in the resected bowels were also examined. Focal staining for pS2 was seen in 32 tumors (51%). In all but one case, less than 10% of the tumor cells were stained. pS2 staining was more common in right sided than in left-sided tumors (p < 0.05), and was more prevalent in Dukes C than combined Dukes A and B tumors (p < 0.05). No significant relationship was found between pS2 positivity and the degree of tumor differentiation, patients' sex, or outcome of the disease as judged by the development of recurrence or metastasis. Strong pS2 positivity was seen in hyperplastic polyps and in nonneoplastic mucosa adjacent to many tumors. Tubular adenomas were either negative or showed focal superficial staining. It is concluded that pS2 immunostaining in colorectal adenocarcinoma does not seem to have prognostic significance, but may reflect developmental differences between the right and left side of the colon. The presence of pS2 staining in adjacent nonneoplastic mucosa and in hyperplastic polyps suggests that the epithelium in these areas is of a regenerative or reactive nature. PMID- 8415591 TI - Nonpapillary and papillary renal cell carcinoma: a cytogenetic and phenotypic study. AB - Cytogenetic and molecular genetic studies allow the common renal cell neoplasms to be separated into two main types: (1) Nonpapillary renal cell carcinomas (RCC) which have a loss of 3p13-pter sequences and (2) Papillary renal cell tumors having tri- or tetrasomies of chromosome 7 and trisomy 17. To investigate renal proximal (PT) and distal (DT) tubular epithelial phenotype expression in these genetically distinct neoplasms, a panel of antibodies and lectins selectively reactive with normal adult PT and DT was applied to 10 nonpapillary and seven papillary RCC. All tumors except one papillary RCC demonstrated characteristic karyotypes. Phenotype expression varied depending upon changes in the histopathologic patterns within a tumor. Among tumors composed of only one cell type, columnar, eosinophilic cells showed only PT staining and small, basophilic cells showed only DT staining. One tumor revealed a transition from small, basophilic cells to columnar, eosinophilic cells. The basophilic cells stained for DT markers and the eosinophilic cells for PT markers. One tumor consisted of nests of clear cells between indistinct papillary structures. The clear cells stained for both PT and DT markers. All 10 nonpapillary RCC demonstrated PT staining; nine exhibited DT markers. Staining was most intense in areas of tumor showing higher nuclear grades, tubuloglandular differentiation or in granular, eosinophilic cells and was absent or weak in solid groups of low nuclear grade clear cells. Papillary and nonpapillary RCC demonstrated lectin-binding or antigens associated with both PT and DT indicating a capacity for multipotential metanephric differentiation in each type of neoplasm.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415592 TI - Immunoreactivities of Reed-Sternberg cells and their variants in the sequential biopsy of Hodgkin's disease. AB - Reactivities of Reed-Sternberg (RS) cells for antibodies against T-lymphocytes and B-lymphocytes together with CD30 and CD15 were examined in the sequential biopsies of Hodgkin's disease (HD). For this purpose, six patients with lymphocyte predominant (LP) HD, each three of nodular and diffuse types, and three with mixed cellularity (MC) HD were examined. Numbers of times of biopsy in these cases were 25, the intervals between each biopsy ranged from 1 to 92 (median 12) mo. Positive reactivities of the RS cells for CD20, 45R, w75, w74, and/or MB1 were observed in all but one with nodular LPHD. Five of six cass with LPHD showed positive reaction for CD30 and/or CD15. RS cells in all of the present MCHD cases showed a positive reaction for CD30 with each one case showing positive reaction for CD15, CD20, and w75. RS cells in two of three cases with MCHD showed a positive reaction for CD20 or w75. RS cells did not show positive reaction for any antibodies against T-lymphocytes. Sequential biopsy revealed that two of three cases with nodular LPHD and one of three with diffuse LPHD changed to MCHD and two of three with MCHD changed to LDHD. Reactivity of RS cells for CD30 was consistent even when the histologic subtypes changed. Expression of CD15 by RS cells was labile; appearance of disappearance of this antigen during sequential biopsy was frequent. Positive reactivities of RS cells for anti-B-lymphocytes antibodies in LPHD cases disappeared in two cases when histology changed to MCHD. In one case with MCHD, the RS cells became reactive for anti-B-lymphocytes antibodies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415593 TI - Aberrant expression of macrophage-associated antigens (CD68 and Ki-M1P) by Schwann cells in reactive and neoplastic neural tissue. Light- and electron microscopic findings. AB - The monoclonal antibodies KP1 (CD68), PG-M1 (CD68), and Ki-M1P can be used to detect normal and neoplastic monocytes/macrophages in formalin-fixed, paraffin embedded tissue. However, systematic investigations undertaken on various tissues have revealed that reactivity with these antibodies is also found in a few cells that do not belong to the mononuclear phagocyte system. The immunoreactivity of normal, reactively altered, and neoplastic Schwann cells with these antibodies was investigated using intact peripheral myelinated nerves, nerves exhibiting Wallerian degeneration, traumatic neuromas, appendixes with neurogenic appendicopathy, granular cell tumors, neurofibromas, and neurogenic sarcomas. The results obtained by light microscopy showed that Schwann cells of nerves with Wallerian degeneration and those in traumatic neuroma, neurofibroma, and granular cell tumor exhibit intracytoplasmic immunoreactivity, which is usually intense, with KP1, Ki-M1P, and PG-M1, but normal myelinated nerves, neurogenic sarcoma, and Schwann cells in neurogenic appendicopathy do not react with these antibodies. No Schwann cells were stained by MAC387 or anti-lysozyme. The site of immunoreactivity with these antibodies was also investigated by electron microscopy. One of the granular cell tumors and macrophages in lymphoid tissue were investigated by the immunogold technique using both pre- and postembedding methods. In granular cell tumor the reaction product was located in phagolysosomes; in macrophages it was found in phagosomes and/or lysosome-like granules. Our findings therefore indicate that immunoreactivity with KP1, Ki-M1P, and PG-M1 can also be expected in cells that do not belong to the mononuclear phagocyte system if they exhibit phagocytosis and/or autophagy. PMID- 8415594 TI - Estrogen and progesterone receptor detection in neoplastic and non-neoplastic thyroid tissues. AB - Previous reports have shown that estrogen and progesterone receptors (ER and PgR) are detectable in neoplastic and non-neoplastic thyroid tissues using a variety of techniques. However, differences in relative expression of ER and PgR according to tumor type have not been emphasized. To determine whether such differences occur and to assess clinicopathologic correlations, we have evaluated ER and PgR expression in sections of 64 thyroid lesions stained by indirect immunoperoxidase. The lesions studied included 39 papillary carcinomas, 3 follicular adenomas, 2 follicular carcinomas, 15 Hurthle cell tumors, 3 medullary carcinomas, and 2 multinodular goiters. Immunostaining was achieved using monoclonal antibodies H222 to ER and JZB39 to PgR. Additionally, the monoclonal antibodies D75p3 to ER and mPRI to PgR were also used in some cases. Overall, ER and/orPgR were detected in 26 of 64 lesions (40%) using any of the above antibodies. ER was identified in eight of 64 (12.5%) tumors that were all papillary carcinomas. No staining for ER was seen in follicular neoplasms, Hurthle cell tumors, or medullary carcinomas. PgR staining was present in 13 of 39 (33%) papillary carcinomas, 8 of 15 (53%) Hurthle cell tumors, 2 of 7 (28%) follicular tumors, and in none of 3 medullary carcinomas. Non-neoplastic thyroid tissue was negative for ER and PgR in all but one case in which weak staining for PgR was present adjacent to a follicular adenoma. There were no correlations between ER and/or PgR staining and age, gender, tumor size, presence of capsular or vascular invasion, or lymph node status in any tumor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415595 TI - Immunostaining with HAM56 in the diagnosis of adenocarcinomas. AB - Data has been presented suggesting that the monoclonal antibody HAM56 reacts with many adenocarcinomas but not with colonic adenocarcinomas. Previous studies have suggested that immunostaining with HAM56 may be useful in the differential diagnosis of adenocarcinoma of unknown origin. To test the specificity of HAM56 immunostaining in the diagnosis of adenocarcinomas, we studied formalin-fixed, paraffin-embedded tissue from 213 adenocarcinomas from a variety of sites using an avidin-biotin immunohistochemical technique. Positive reactivity was seen in some tumors of all types including 39% of colonic adenocarcinomas. We conclude that immunostaining with HAM56 is not specific in the differential diagnosis of adenocarcinoma of unknown origin. PMID- 8415596 TI - Research by pathologists in the United States: analysis of publications. AB - Academic pathology is a unique profession in which the ability to pursue almost any type of academic or scholarly endeavor can be supported. Although it is difficult to define methods of comparing individual achievements in medicine, for investigative activity, common measures of academic productivity are numbers of peer-reviewed publications and National Institute of Health support. Articles from pathology departments were randomly selected from MEDLINE. One hundred and five articles were examined for the following features: financial support, human subjects, the presence of a photomicrograph, molecular biology techniques, and the number of times each article was cited as determined using the Science Citation Index. The articles were assigned to one of the following categories: anatomical pathology, clinical pathology, or research. The following percentages of articles were federally funded: total, 43.8; anatomical pathology, 33.3; clinical pathology, 9.5 and research, 74.4. The citations per article for federally funded and not federally funded articles, respectively, for each as these categories was as follows: total, 18.7 and 5.3; anatomical pathology, 15.7 and 5.5; clinical pathology, 2.0 and 4.9; and research, 21.4 and 5.7. It can be concluded from the data presented that federally funded articles from departments of pathology are cited more often than articles that are not federally funded. The most impressive finding in reviewing the articles is the wide diversity of research conducted in pathology departments. The use of federal funding or citation analysis to evaluate the performance of an individual pathologist would be a mistake. PMID- 8415597 TI - Congenital peribronchial myofibroblastic tumor (so-called "congenital leiomyosarcoma"). A distinct neonatal lung lesion associated with nonimmune hydrops fetalis. AB - An unusual solid tumor composed of myofibroblasts in an infant with nonimmune hydrops fetalis is presented together with a review of previous cases reported under a variety of different names, most notably congenital leiomyosarcoma. Immunocytochemical, ultrastructural, and flow cytometric data plus an analysis of fetal lung specimens from various times during lung development suggest that this tumor arises from the condensed mesenchyme that surrounds those respiratory ducts that go on to form large bronchi. Because of its age of onset, cellular composition, and benign biological behavior we believe that this tumor is analogous to two other congenital myofibroblastic tumors, congenital mesoblastic nephroma, and spindle cell tumor of the intestinal tract. Although the term myofibroblastoma has gained popularity in recent reports describing localized proliferations of myofibroblasts in adults, we would like to emphasize the unique developmental origin of this lesion and suggest the term congenital peribronchial myofibroblastic tumor. PMID- 8415598 TI - Calcium oxalate crystal deposition in necrotizing otomycosis caused by Aspergillus niger. AB - Numerous calcium oxalate crystals were present within fruiting heads of Aspergillus niger and among necrotic debris in a case of bilateral invasive otomycosis occurring in a diabetic female with end stage renal disease. This is the first report of in vivo calcium oxalate crystal deposition associated with Aspergillus niger at this anatomic site. The presence of localized oxalate crystals within necrotic tissue from the external auditory canal is presumptive of otomycosis caused by Aspergillus niger, and may serve as a diagnostic clue to the etiologic agent before histologic demonstration of hyphae or growth in culture. PMID- 8415599 TI - Cellular angiolipoma of the breast. AB - Cellular angiolipomas are benign fatty tumors that occur as multiple subcutaneous nodules on the extremities and trunks of young adults. Although clinically benign, they may occasionally mimic Kaposi's sarcoma or angiosarcoma histologically. We report the first case of cellular angiolipoma occurring in the subcutaneous tissue of the breast, a rare but well-recognized site of benign and malignant vascular tumors. PMID- 8415600 TI - Demonstration of coexistent B-cell lymphoma and therapy-related acute myelogenous leukemia in lymph nodes by flow cytometry and immunohistochemistry. Case report and review of the literature. AB - A 58-yr-old white woman, with a 9-yr history of non-Hodgkin's lymphoma with recurrences and multi-modality therapy, presents with acute leukemia and residual lymphadenopathy. By flow cytometric analysis, cytochemistry, and cytomorphology, the leukemia is classified as acute myelogenous leukemia (AML), M4, and the lymph nodes are found to contain residual NHL in addition to an AML infiltrate. Immunohistochemistry performed on the lymph nodes correlates with the flow cytometric data. A review of the literature of AML secondary to therapy for non Hodgkin's lymphoma is also conducted. There are no documented cases in the English literature of simultaneous involvement of lymph nodes by residual lymphoma and an AML infiltrate. Thus, this report is unique in the simultaneous involvement of lymph nodes by residual lymphoma and therapy-related AML. It emphasizes the application and usefulness of flow cytometry and immunohistochemistry in such cases. PMID- 8415601 TI - Correspondence re: A. G. Nerlich, S. Majewski, N. Hunzelmann, R. E. Brenner, B. Wiebecke, P. K. Muller, T. Krieg, and K. Remberger. Distinctive case. Excessive collagen formation in fibrolamellar carcinoma of the liver: a morphological and biochemical study. Mod Pathol 5:580, 199. PMID- 8415602 TI - High titers of retrovirus (vesicular stomatitis virus) pseudotypes, at last. PMID- 8415603 TI - DNA methylation: a phoenix rises. PMID- 8415604 TI - Molecular analysis of the microtubule motor dynein. AB - Dynein is a large enzyme complex that has been found in recent years to be responsible for a variety of forms of intracellular movement associated with microtubules. Molecular analysis of several of the polypeptide components of dynein and a related complex has provided important new insight into their structural organization and mechanism of action in the cell. PMID- 8415605 TI - Tissue culture-induced DNA methylation variation in maize. AB - Twenty-one progeny lines derived from tissue cultures of two embryo sources of maize inbred strain A188 were examined for DNA methylation changes. Total DNA was cut with the isoschizomers Hpa II and Msp I and probed with 18 single-copy Pst I genomic clones and two cDNA clones. Eight of these probes could detect both increases and decreases in methylation. With these probes 39% of the families were found to contain an altered methylation pattern. All changes represented a decrease in methylation. The other 12 probes could detect only increases in methylation; no methylation variation was seen with these probes. Fifteen percent of the methylation changes were homozygous in the original regenerated plant. Changes were stably inherited upon two generations of self-pollination. No sequence variation was observed in Msp I-digested DNA from the same 21 progeny lines. Certain probes detected methylation changes much more often than others. Our study provides evidence that demethylation occurs at a high frequency and could be an important cause of tissue culture-induced variation. Occurrence of the frequent homozygous alterations in original regenerated plants implies a non random mutational mechanism. PMID- 8415606 TI - Weighting in sequence space: a comparison of methods in terms of generalized sequences. AB - Four methods for weighting aligned biological sequences have recently appeared that differ mathematically, philosophically, and in their results. Thus, while there is consensus about the need to weight sequences, the method to use is contentious. A geometric analysis based on a continuous sequence space is presented that provides a common framework in which to compare the methods. It is concluded that there are two "best" methods. When the sequences are known to be phylogenetically related and a tree can be generated without introducing excessive stress into the data, the method of Altschul et al. [Altschul, S. F., Carroll, R. J. & Lipman, D. J. (1989) J. Mol. Biol. 207, 647-653] is appropriate. When the sequences are not known to be phylogenetically related or a tree cannot be produced without unduly distorting the distances between the sequences, a modification of the method of Sibbald and Argos [Sibbald, P. R. & Argos, P. (1990) J. Mol. Biol. 216, 813-818] is preferable. PMID- 8415607 TI - The disaggregation theory of signal transduction revisited: further evidence that G proteins are multimeric and disaggregate to monomers when activated. AB - We have compared the sedimentation rates on sucrose gradients of the heterotrimeric GTP-binding regulatory (G) proteins Gs, G(o), Gi, and Gq extracted from rat brain synaptoneurosomes with Lubrol and digitonin. The individual alpha and beta subunits were monitored with specific antisera. In all cases, both subunits cosedimented, indicating that the subunits are likely complexed as heterotrimers. When extracted with Lubrol all of the G proteins sedimented with rates of about 4.5 S (consistent with heterotrimers) whereas digitonin extracted 60% of the G proteins with peaks at 11 S; 40% pelleted as larger structures. Digitonin-extracted Gi was cross-linked by p-phenylenedimaleimide, yielding structures too large to enter polyacrylamide gels. No cross-linking of Lubrol extracted Gi occurred. Treatment of the membranes with guanosine 5'-[gamma thio]triphosphate and Mg2+ yielded digitonin-extracted structures with peak sedimentation values of 8.5 S--i.e., comparable to that of purified G(o) in digitonin and considerably larger than the Lubrol-extracted 2S structures representing the separated alpha and beta gamma subunits formed by the actions of guanosine 5'-[gamma-thio]triphosphate. It is concluded that the multimeric structures of G proteins in brain membranes are at least partially preserved in digitonin and that activation of these structures in membranes yields monomers of G proteins rather than the disaggregated products (alpha and beta gamma complexes) observed in Lubrol. It is proposed that hormones and GTP affect the dynamic interplay between multimeric G proteins and receptors in a fashion analogous to the actions of ATP on the dynamic interactions between myosin and actin filaments. Signal transduction is mediated by activated monomers released from the multimers during the activation process. PMID- 8415608 TI - Phosphorylation-dependent binding of a signal molecule to the flagellar switch of bacteria. AB - Regulation of the direction of flagellar rotation is central to the mechanism of bacterial chemotaxis. The transitions between counterclockwise and clockwise rotation are controlled by a "switch complex" composed of three proteins (FliG, FliM, and FliN) and located at the base of the flagellar motor. The mechanism of function of the switch is unknown. Here we demonstrate that the diffusible clockwise-signal molecule, the CheY protein, binds to the switch, that the primary docking site is FliM, that the extent of CheY binding to FliM is dependent upon the phosphorylation level of CheY, and that it is unaffected by the other two switch proteins. This study provides a biochemical demonstration of binding of a signal molecule to the bacterial switch and demonstrates directly that phosphorylation regulates the activity of this molecule. PMID- 8415609 TI - Physiological compensation in antisense transformants: specific induction of an "ersatz" glucan endo-1,3-beta-glucosidase in plants infected with necrotizing viruses. AB - Plant class I glucan endo-1,3-beta-glucosidases (beta-1,3-glucanase; 1,3-beta-D glucan glucanohydrolase, EC 3.2.1.39) have been implicated in development and defense against pathogen attack. Nevertheless, beta-1,3-glucanase deficiencies generated by antisense transformation of Nicotiana sylvestris and tobacco have little biological effect. We report here that another beta-1,3-glucanase activity is induced in these deficient mutants after infection with necrotizing viruses. Induction of class I beta-1,3-glucanase was markedly inhibited in leaves of N. sylvestris and tobacco antisense transformants infected with tobacco necrosis virus and tobacco mosaic virus, respectively. A serologically distinct beta-1,3 glucanase activity was present in the infected antisense transformants but was absent in both healthy and infected control plants and in antisense transformants treated with the stress hormone ethylene. Immunoblot analyses, localization studies, and measurements of antibody specificity indicate that this compensatory beta-1,3-glucanase activity is an intracellular enzyme different from known tobacco beta-1,3-glucanases. Therefore, plants can compensate for a deficiency in enzyme activity by producing a functionally equivalent replacement--i.e., "ersatz"--protein or proteins. The fact that compensation for beta-1,3-glucanase activity occurs in response to infection argues strongly for an important role of these enzymes in pathogenesis. PMID- 8415610 TI - Differential usage of multiple brain-derived neurotrophic factor promoters in the rat brain following neuronal activation. AB - The rat brain-derived neurotropic factor (BDNF) gene consists of four 5' exons linked to separate promoters and one 3' exon encoding the prepro-BDNF protein. To gain insights into the regulation of BDNF mRNA expression, probes specific for the different 5' exons were used to study the expression of BDNF mRNA in the brain. Following a systemic injection of the glutamate analog kainic acid, exon I, II, and III mRNAs increased transiently in hippocampus and cerebral cortex. A modest increase was seen for exon IV, where a new transcription initiation site was induced by this treatment. Pretreatments with the N-methyl-D-aspartate (NMDA) receptor antagonist MK801 or the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor antagonist 2,3-dihydroxy-6 nitrosulfanoylbenzo(f)quinoxaline revealed two region-specific patterns of glutamate receptor-mediated regulation. The first pattern found in neocortex, piriform cortex, and amygdala involves regulation of BDNF exon I, II, and III mRNAs through NMDA and AMPA/kainate receptors. The second pattern found in the hippocampus involves regulation of BDNF exon I, II, and III mRNAs by high affinity kainate or metabotropic receptors. Treatment with the gamma-aminobutyric acid subtype A (GABAA) receptor antagonist bicuculline increased exon I and III mRNAs in the denate gyrus, and the muscarinic receptor agonist pilocarpine increased exon I mRNA mainly in the neocortex. These data show that the four BDNF promoters allow multiple points of BDNF mRNA regulation and suggest that the activation of different subtypes of glutamate receptors differentially regulates the expression of BDNF exon-specific mRNAs in the brain. PMID- 8415611 TI - Molecular cloning of the gene for the allatostatin family of neuropeptides from the cockroach Diploptera punctata. AB - Allatostatins (ASTs) are insect neuropeptides that inhibit juvenile hormone biosynthesis by the corpora allata. We have isolated a cDNA from the cockroach Diploptera punctata that encodes a 41.5-kDa precursor polypeptide containing the AST family of peptides. Translation of the cDNA revealed a 370-amino acid pre-pro peptide consisting of 13 AST-type peptides and appropriate processing sites for endoproteolytic cleavage and amidation. The 13 potential AST sequences are characterized by the C-terminal AST corestructure Phe-Gly-Leu-NH2, with only one exception. Separating the clustered ASTs in the precursor, three acidic spacer regions are found. Contained within the largest of these are two potentially related peptides that may also be processed. Southern blot analysis revealed the presence of a single copy of the AST gene per haploid genome, as well as the probability that the gene may be present in at least two allelic forms. In situ hybridization indicated the AST-encoding gene is expressed in neurosecretory cells of D. punctata brain. PMID- 8415612 TI - Growth of the cyanobacterium Anabaena on molecular nitrogen: NifJ is required when iron is limited. AB - The nifJ gene of Klebsiella pneumoniae encodes an oxidoreductase required for the transfer of electrons from pyruvate to flavodoxin, which reduces nitrogenase. The nifJ gene of Anabaena 7120, isolated from a cosmid bank, was found to contain an open reading frame encoding a 1197-aa protein. The deduced amino acid sequence shows 50% identity to the Klebsiella homolog. The nifJ gene in Anabaena 7120 was inactivated by chromosomal interruption. The resulting mutant was unable to grow on medium depleted of both iron and combined nitrogen but grew normally, fixing nitrogen, when iron was present. NifJ transcripts of 2.7 and 4.3 kb are induced by iron depletion irrespective of nitrogen status. One particular stretch of the Anabaena 7120 nifJ gene encodes 12 aa with no complementary matches in the Klebsiella protein. This insert contains five tandem repeats of the heptamer CCCCAGT. These heptamers, as well as heptamers and octamers of other related sequences, have been located in a number of cyanobacterial genomes but are usually not found within the coding region of a gene. The site of the Anabaena 7120 heptamers in the nifJ genes of other filamentous cyanobacteria contains a surprising diversity of repeated sequences, both octamers and heptamers. The corresponding protein inserts range in length from 1 to 21 aa, relative to Klebsiella NifJ. PMID- 8415613 TI - Transgenic mouse model of malignant skin melanoma. AB - Tyr-SV40E transgenic mice are specifically susceptible to melanoma due to expression of the oncogene in pigment cells. Mice of the more susceptible lines die young of early-onset eye melanomas, when skin melanomas are still infrequent and benign. To surmount this obstacle, skin from donors of two high susceptibility lines was grafted to Tyr-SV40E hosts of a low-susceptibility line of the same inbred strain, thereby enabling the skin to outlive the donors and continue to grow in immunocompetent but tolerant hosts. Unexpectedly, donor pigment cells in all the grafts soon selectively proliferated close to areas of greatest wound healing, forming a dense black tracery, especially at the outer rim of the grafts. These lesions slowly grew radially within the grafts, producing irregular greyish patches. Local vertical thickenings then appeared and developed into small melanomas, which soon ulcerated through the epidermis. The tumors rapidly enlarged and became deeply invasive. Discrete black nevi also arose, with many becoming larger and distinctly blue, but those not near areas of pronounced wound healing did not progress to malignancy. In this first series, malignant melanoma resulted in all the grafts from the more susceptible of two donor lines and in some grafts from the other line. Distant metastases occurred in some cases from each line. Most tumors were hypomelanotic and heterogeneous, with lobes or areas differing in melanization. The results strongly suggest that growth factors and cytokines--known to be produced in wound repair--are triggering the growth and malignant conversion of these genetically susceptible melanocytes and that in the graft situation we are merely witnessing a caricature -a usefully exaggerated manifestation of the true events underlying the genesis of melanomas. The striking resemblance to the human malignancy, the genetic uniformity and different susceptibilities of the transgenic lines, and the experimental possibilities in the grafted mice all make them an excellent model of the disease. PMID- 8415614 TI - Histopathogenesis of malignant skin melanoma induced in genetically susceptible transgenic mice. AB - Animal models of human malignant skin melanoma were created in melanoma susceptible inbred-strain transgenic mice by grafting skin from donors of high susceptibility lines to hosts of a low-susceptibility line, thereby overcoming the problem of early death of the more susceptible animals from eye melanomas. As already described [Mintz, B. & Silvers, W. K. (1993) Proc. Natl. Acad. Sci. USA 90, 8817-8821], melanocytes within the grafts selectively proliferated in close proximity to areas of greatest wound healing, presumably in response to mitogenic factors from cells contributing to wound repair. An orderly sequence of externally visible events culminated in malignant melanoma. We examine here the histogenetic concomitants of these changes and find that they define a stepwise sequence strikingly comparable to that leading to human cutaneous melanoma. Moreover, the histological details suggest some of the underlying mechanisms. While the early lesions are first seen in the superficial dermis in the mouse, and in the basal layer of the epidermis in the human, both progress by radial growth followed by vertical growth. Melanocytic hyperplasia resulted in nests of densely melanized fusiform cells which were losing their dendrites. Some discrete lesions in the deep dermis appeared as blue nevi. As radial proliferation advanced, cellular atypia increased and the previously independent melanocytes cohered closely and formed a small solid tumor; the cells were usually then hypomelanotic or amelanotic. Ulceration of tumor through the epidermis occurred early. The tumor mass grew rapidly in the deep dermis and invaded and destroyed subcutaneous tissue and muscle. Primary tumors in the skin were often heterogeneous, with lobules or regions differing in pigmentation or atypia. However, the cells in circulating emboli, or in metastases in lymph nodes and lungs, appeared relatively homogeneous. These genetically uniform transgenic mouse models provide experimental access to the multistage genesis of melanoma. PMID- 8415615 TI - Protein phosphatase inhibitors induce the selective breakdown of stable microtubules in fibroblasts and epithelial cells. AB - In many cell types, a small subset of microtubules (MTs) are unusually long-lived compared with the majority of the MTs. These "stable" MTs may be important mediators of differentiative events since they are usually found aligned with developing asymmetries of cells undergoing morphogenesis. In addition to their longevity, the stable MTs are more resistant to drug depolymerization and are enriched in post-translationally detyrosinated tubulin (Glu-tubulin). To determine the role of protein phosphorylation in the regulation of these stable MTs, we treated NIH 3T3 fibroblasts and TC-7 monkey kidney epithelial cells with okadaic acid (OA) and calyculin A, potent inhibitors of protein phosphatases 1 and 2A (PP1 and PP2A), and then localized dynamic MTs and stable MTs with antibodies specific for tyrosinated tubulin (Tyrtubulin) and Glu-tubulin, respectively. OA at 0.1-10 microM caused a rapid and complete breakdown of Glu MTs (MTs enriched in Glu-tubulin) in both cell types without substantially affecting the number of Tyr-MTs. While all concentrations of OA over this range resulted in a complete loss of Glu-MTs, the onset of Glu-MT breakdown was proportional to the logarithm of the OA concentration. The inactive analog of OA, 1-norokadaone, had no effect at any concentration. Calyculin A also caused a selective loss of Glu-MTs but was effective at 10 nM, consistent with its more potent inhibition of PP1. That the loss of Glu-MTs reflected the loss of stable MTs from the cells was shown by the absence of nocodazole-resistant MTs in OA treated cells. OA did not appear to activate a MT-severing activity, since no MT fragments were observed after OA treatment of cells pretreated with taxol. These results suggest that PP1 and perhaps PP2A are involved in the regulation of MT stability in cells and show that the dynamic and stable subsets of MTs are regulated differentially by protein phosphorylation. PMID- 8415616 TI - Interaction of human thyroid hormone receptor beta with transcription factor TFIIB may mediate target gene derepression and activation by thyroid hormone. AB - The human thyroid hormone receptor beta (hTR beta) is capable of both transcriptional silencing and hormone-dependent activation. However, the detailed mechanism of this transcriptional regulation remains to be elucidated. One possibility is that hTR beta interacts directly with factors of the basal transcriptional machinery, thereby modulating basal promoter activity in a direct manner, as has been shown for other transcription factors. Here, we show that hTR beta interacts specifically with the human basal transcription factor TFIIB. Deletion analysis revealed two contact sites in the receptor: one is located in the N terminus, while the other is part of the ligand-binding domain (LBD) and is located at the C terminus. Interestingly, each receptor contact site interacts with different sites in TFIIB. Cotransfection experiments revealed that, when fused to the DNA-binding domain of yeast transcription factor GAL4, the C terminal interaction site of hTR beta was transcriptionally inactive; however, when it was cotransfected with the remaining part of the LBD on a separate molecule, silencing function was restored. In agreement with that, we show that thyroid hormone is able to significantly decrease the interaction of its receptor LBD with TFIIB. Our data suggest that hTR beta acts as a transcriptional silencer by interacting with TFIIB and that thyroid hormone may act in part by preventing transcriptional repression at this level. PMID- 8415617 TI - Identification of an essential gene, l(3)73Ai, with a dominant temperature sensitive lethal allele, encoding a Drosophila proteasome subunit. AB - Proteasomes are multicatalytic proteinase complexes that function as a major nonlysosomal proteolytic system in all eukaryotes. These particles are made up of 13-15 nonidentical subunits, and they exhibit multiple endopeptidase activities that promote the intracellular turnover of abnormal polypeptides and short-lived regulatory proteins. Although the biochemical characterization of proteasomes has been quite extensive, and although a number of the genes encoding proteasome subunits have been cloned from various organisms, there is still much to be learned about their function in vivo and what role(s) they might play during development. Here, we report the identification of the l(3)73Ai1 allele of Drosophila melanogaster as a dominant temperature-sensitive lethal mutation in a gene encoding a component of the proteasome, thus opening the way for future genetic and developmental studies on this important proteolytic system in a higher eukaryote. PMID- 8415618 TI - Physical association between the high-affinity IgG receptor (Fc gamma RI) and the gamma subunit of the high-affinity IgE receptor (Fc epsilon RI gamma). AB - To investigate the structural basis of transmembrane signaling via the high affinity IgG receptor (Fc gamma RI), the identity of Fc gamma RI-associated proteins in THP-1 human monocytic cells was examined. Anti-Fc gamma RI monoclonal antibody (mAb) 197 immunoprecipitates from 125I-labeled THP-1 cells solubilized in 1% digitonin buffer were found to contain a protein migrating at 12 kDa on reduction. This protein comigrated with the 12-kDa protein precipitated by the anti-high-affinity IgE receptor gamma chain (Fc epsilon RI gamma) mAb 4D8. Similarly, a 70-kDa band immunoprecipitated by mAb 4D8 comigrated with the 70-kDa protein band corresponding to Fc gamma RI in mAb 197 immunoprecipitates. On two dimensional nonreducing-reducing gel analysis, the 12-kDa protein present in both mAb 197 and mAb 4D8 immunoprecipitates migrated as a disulfide-linked homodimer. Analysis of 1% Nonidet P-40 eluates of the digitonin immunoprecipitates under reducing conditions demonstrated the presence of a 12-kDa band in the mAb 197 immunoprecipitate that could be reprecipitated with mAb 4D8. Conversely, a 70-kDa band in the mAb 4D8 immunoprecipitate could be reprecipitated with mAb 197. Similar to these findings, both mAb 197 and mAb 4D8 precipitated a 12-kDa disulfide-linked homodimeric protein from digitonin lysates of 125I-labeled human neutrophils after induction of Fc gamma RI expression with interferon gamma but not from unstimulated neutrophils. Northern blot analysis confirmed the presence of Fc epsilon RI gamma mRNA in interferon gamma-induced human neutrophils. We conclude that Fc epsilon RI gamma, a member of a family of proteins implicated in transmembrane signaling via immune recognition receptors, associates with Fc gamma RI in human cells. PMID- 8415619 TI - Loss of resistance to dietary cholesterol in the rat after hypophysectomy: importance of the presence of growth hormone for hepatic low density lipoprotein receptor expression. AB - This investigation was undertaken to determine the role of pituitary function and, in particular, the possible influence of growth hormone (GH) on hepatic low density lipoprotein (LDL)-receptor expression in response to dietary cholesterol. Feeding normal rats with 2% cholesterol for 5 or 6 days did not alter LDL receptor numbers, LDL-receptor mRNA levels, or plasma cholesterol, although hepatic cholesterol increased 5-fold. When hypophysectomized rats received the same diet, the LDL-receptor number and its mRNA levels were reduced by 75%, plasma cholesterol increased 6-fold, and hepatic cholesterol increased 12-fold. Stepwise hormonal substitution of cholesterol-fed, hypophysectomized rats revealed that substitution with GH was important to restore hepatic LDL-receptor number and mRNA levels. The presence of GH was also important to reduce the hypercholesterolemia in cholesterol-fed hypophysectomized rats. We conclude that the presence of GH is important for hepatic LDL-receptor expression, both at the protein and the mRNA level. The resistance to suppression of rat hepatic LDL receptors by dietary cholesterol depends, at least in part, on the presence of GH. PMID- 8415620 TI - Identification of a receptor for gamma melanotropin and other proopiomelanocortin peptides in the hypothalamus and limbic system. AB - Corticotropin (ACTH) and melanotropin (MSH) peptides (melanocortins) are produced not only in the pituitary but also in the brain, with highest concentrations in the arcuate nucleus of the hypothalamus and the commisural nucleus of the solitary tract. We have identified a receptor for MSH and ACTH peptides that is specifically expressed in regions of the hypothalamus and limbic system. This melanocortin receptor (MC3-R) is found in neurons of the arcuate nucleus known to express proopiomelanocortin (POMC) and in a subset of the nuclei to which these neurons send projections. The MC3-R is 43% identical to the MSH receptor present in melanocytes and is strongly coupled to adenylyl cyclase. Unlike the MSH or ACTH receptors, MC3-R is potently activated by gamma-MSH peptides, POMC products that were named for their amino acid homology with alpha- and beta-MSH, but lack melanotropic activity. The primary biological role of the gamma-MSH peptides is not yet understood. The location and properties of this receptor provide a pharmacological basis for the action of POMC peptides produced in the brain and possibly a specific physiological role for gamma-MSH. PMID- 8415621 TI - Localization of D1 and D2 dopamine receptors in brain with subtype-specific antibodies. AB - Five or more dopamine receptor genes are expressed in brain. However, the pharmacological similarities of the encoded D1-D5 receptors have hindered studies of the localization and functions of the subtypes. To better understand the roles of the individual receptors, antibodies were raised against recombinant D1 and D2 proteins and were shown to bind to the receptor subtypes specifically in Western blot and immunoprecipitation studies. Each antibody reacted selectively with the respective receptor protein expressed both in cells transfected with the cDNAs and in brain. By immunocytochemistry, D1 and D2 had similar regional distributions in rat, monkey, and human brain, with the most intense staining in striatum, olfactory bulb, and substantia nigra. Within each region, however, the precise distributions of each subtype were distinct and often complementary. D1 and D2 were differentially enriched in striatal patch and matrix compartments, in selective layers of the olfactory bulb, and in either substantia nigra pars compacta or reticulata. Electron microscopy demonstrated that D1 and D2 also had highly selective subcellular distributions. In the rat neostriatum, the majority of D1 and D2 immunoreactivity was localized in postsynaptic sites in subsets of spiny dendrites and spine heads in rat neostriatum. Presynaptic D1 and D2 receptors were also observed, indicating both subtypes may regulate neurotransmitter release. D1 was also present in axon terminals in the substantia nigra. These results provide a morphological substrate for understanding the pre- and postsynaptic functions of the genetically defined D1 and D2 receptors in discrete neuronal circuits in mammalian brain. PMID- 8415622 TI - Chimeric-transgenic mice represent a powerful tool for studying how the proliferation and differentiation programs of intestinal epithelial cell lineages are regulated. AB - An in vivo system has been developed for examining the effects of wild-type or mutant proteins on cell fate determination in the mouse intestinal epithelium or on the proliferation and differentiation programs of its component epithelial lineages. This system takes advantage of the fact that at the conclusion of gut morphogenesis, each intestinal crypt is composed of a monoclonal population of cells descended from a single active multipotent stem cell, each villus is supplied by several monoclonal crypts, and the four principal cell types of the intestinal epithelium differentiate during a rapid, geographically well-organized migration along the crypt-to-villus axis. Embryonic stem (ES) cells (129/Sv origin) are initially transfected with recombinant DNAs consisting of a reporter of interest linked to transcriptional regulatory elements that control the cell lineage-specific, differentiation-dependent, and axial patterns of expression of fatty acid binding protein genes in the gut. Stably transfected ES cells are subsequently introduced into host C57BL/6 blastocysts to generate chimeric transgenic mice. At the borders of ES cell-derived and host blastocyst-derived epithelium, intestinal villi are found that are supplied by both ES cell- and host blastocyst-derived crypts. These villi can be rapidly identified in fixed whole-mount preparations of intestine using the alpha-L-fucose-specific Ulex europaeus agglutinin type I (UEA-I) lectin. They appear striped because UEA-I recognizes a cell-surface carbohydrate polymorphism between the inbred strains used to generate the chimeric animals. The strength of this system derives from the fact that two gut epithelial populations can be compared and contrasted that occupy virtually identical positions along the crypt-to-villus and duodenal-to colonic axes within the same animal and differ only by the presence or absence of a single gene product. The band of blastocyst-derived epithelium in these striped, polyclonal villi can be used as an internal control to assess the biological effect of the transfected gene product produced in the adjacent stripe of ES-derived cells. The system can be used for either gain-of-function or loss of-function experiments. PMID- 8415623 TI - Cell lineage-specific and differentiation-dependent patterns of CCAAT/enhancer binding protein alpha expression in the gut epithelium of normal and transgenic mice. AB - The proliferation and differentiation programs of gut epithelial cells are expressed rapidly and perpetually along an anatomically well defined pathway. The mouse intestine thus provides an excellent in vivo model system to define the contributions of CCAAT enhancer binding protein alpha (C/EBP alpha) and related bZIP proteins to these processes. Immunocytochemical studies revealed that C/EBP alpha is produced in villus-associated enterocytes located in the duodenum and jejunum of adult mice. The protein is located in the cytoplasmic and nuclear compartments of these cells. C/EBP alpha is not detectable in proliferating and nonproliferating epithelial cells situated in small intestinal crypts nor is it evident in any gut epithelial cell lineage located in the ileum and colon. The related C/EBP beta and C/EBP delta proteins are not detectable by sensitive immunocytochemical methods in epithelial cells distributed along the duodenal-to colonic axis. Developmental surveys indicate that C/EBP alpha is confined to postmitotic, villus-associated epithelial cells during conversion of the polyclonal intervillus epithelium to monoclonal crypts. Analyses of intestinal isografts reveal that these developmental stage-specific, lineage-specific, differentiation-dependent, and regional patterns of C/EBP alpha expression can be established and maintained in the absence of exposure to luminal contents. Transgenic mice containing nucleotides -1178 to +28 of the rat intestinal fatty acid binding protein gene (I-FABP-1178 to +28) linked to the simian virus 40 large tumor antigen (T antigen) gene express T antigen in villus-associated enterocytes. This results in reentry of enterocytes into the cell cycle and a silencing of C/EBP alpha expression without an apparent effect on the accumulation of several markers of this lineage's terminal differentiation program or on gut morphogenesis. These findings indicate that there is a relationship between expression of C/EBP alpha in enterocytes and their exit from the cell cycle and suggest that I-FABP-1178 to +28/simian virus 40 T antigen transgenic mice could provide a screening assay for examining the role of C/EBP alpha in regulating the activity of genes known to be transcribed during differentiation of this gut epithelial cell lineage. PMID- 8415624 TI - Monoclonal anti-idiotypic antibodies as functional internal images of enzyme active sites: production of a catalytic antibody with a cholinesterase activity. AB - Monoclonal antibody 9A8 was selected by immunizing mice with AE-2, a monoclonal antibody directed against the active site of acetylcholinesterase. In accordance with the idiotypic network theory, monoclonal anti-idiotypic antibody 9A8 displayed internal-image properties of the original immunogen, the acetylcholinesterase active site. Hydrolysis of acetylthiocholine and related esters of thiocholine by 9A8 follows saturation kinetics and kinetic parameters were determined. The hydrolytic activity is characterized by a lowered kcat value (81 s-1) and an increased Km value (0.6 mM) when compared with the original enzyme. However, the rate acceleration (kcat/kuncat = 4.15 x 10(8) remains higher than for the esterase activities usually described for catalytic antibodies directed against transition-state analogs. The 9A8 activity exhibits a relaxation of specificity toward both substrates and inhibitors. This specificity does not correspond to a known enzymatic activity. The anti-idiotypic approach should be valuable for producing different structural and functional copies of the same enzyme active site. This should allow further insights into structure-activity relationships. Furthermore, use of chemically modified enzymes as immunogens may result in anti-idiotypic antibodies with catalytic activities not found in the native enzymes. PMID- 8415625 TI - The amino-terminal 200 amino acids of the plasma membrane Na+,K+-ATPase alpha subunit confer ouabain sensitivity on the sarcoplasmic reticulum Ca(2+)-ATPase. AB - Cardiac glycosides such as G-strophanthin (ouabain) bind to and inhibit the plasma membrane Na+,K(+)-ATPase but not the sarcoplasmic reticulum (SR) Ca(2+) ATPase, whereas thapsigargin specifically blocks the SR Ca(2+)-ATPase. The chimera [n/c]CC, in which the amino-terminal amino acids Met1 to Asp162 of the SR Ca(2+)-ATPase (SERCA1) were replaced with the corresponding portion of the Na+,K(+)-ATPase alpha 1 subunit (Met1 to Asp200), retained thapsigargin- and Ca(2+)-sensitive ATPase activity, although the activity was lower than that of the wild-type SR Ca(2+)-ATPase. Moreover, this Ca(2+)-sensitive ATPase activity was inhibited by ouabain. The chimera NCC, in which Met1-Gly354 of the SR Ca(2+) ATPase were replaced with the corresponding portion of the Na+,K(+)-ATPase, lost the thapsigargin-sensitive Ca(2+)-ATPase activity seen in CCC and [n/c]CC. [3H]Ouabain binding to [n/c]CC and NCC demonstrated that the affinity for this inhibitor seen in the wild-type chicken Na+,K(+)-ATPase was restored in these chimeric molecules. Thus, the ouabain-binding domains are distinct from the thapsigargin sites; ouabain binds to the amino-terminal portion (Met1 to Asp200) of the Na+,K(+)-ATPase alpha 1 subunit, whereas thapsigargin interacts with the regions after Asp162 of the Ca(2+)-ATPase. Moreover, the amino-terminal 200 amino acids of the Na+,K(+)-ATPase alpha 1 subunit are sufficient to exert ouabain dependent inhibition even after incorporation into the corresponding portion of the Ca(2+)-ATPase, and the segment Ile163 to Gly354 of the SR Ca(2+)-ATPase is critical for thapsigargin- and Ca(2+)-sensitive ATPase activity. PMID- 8415626 TI - Testis-specific expression of a metallothionein I-driven transgene correlates with undermethylation of the locus in testicular DNA. AB - Mice carrying a chimeric transgene of the human testis-specific lactate dehydrogenase cDNA driven by mouse metallothionein I promoter have been reported to express the transgene in a testis-specific manner in six founder lines. To study the mechanism by which this testis-specific expression is mediated, we have examined genomic placement, expression pattern, and methylation status of the transgene. Our results indicate that transgene expression is repressed in all somatic tissues examined even when heavy metals are administered. Nuclear run-on assays indicate that failure of expression in the liver (in which the metallothionein I promoter is highly active) occurs at the transcriptional level. In contrast, the transgene mRNA is transcribed in male germ cells and is developmentally regulated during spermatogenesis. Examination of the transgene methylation status reveals that expression is inversely correlated with hypermethylation of the locus; all CpG dinucleotides examined in the promoter region were found to be fully methylated in kidney and liver but were undermethylated in testis. Since methylation of the murine metallothionein I promoter is sufficient to inhibit its activity, it is likely that suppression of the transgene in somatic tissues is mediated by methylation. PMID- 8415627 TI - Expression of an exogenous eukaryotic DNA methyltransferase gene induces transformation of NIH 3T3 cells. AB - Abnormal regional increases in DNA methylation, which have potential for causing gene inactivation and chromosomal instability, are consistently found in immortalized and tumorigenic cells. Increased DNA methyltransferase activity, which is also a characteristic of such cells, is a candidate to mediate these abnormal DNA methylation patterns. We now show that, in NIH 3T3 mouse fibroblasts, constitutive overexpression of an exogenous mouse DNA methyltransferase gene results in a marked increase in overall DNA methylation which is accompanied by tumorigenic transformation. These transformation changes can also be elicited by dexamethasone-inducible expression of an exogenous DNA methyltransferase gene. Our findings provide strong evidence that the increase in DNA methyltransferase activity associated with tumor progression could be a key step in carcinogenesis and provide a model system that can be used to further study this possibility. PMID- 8415628 TI - A second transcriptionally active DNA-binding site for the Wilms tumor gene product, WT1. AB - The putative Wilms tumor suppressor gene, wt1, encodes a zinc-finger protein that binds to the DNA sequence 5'-GCGGGGGCG-3'. We previously reported that WT1 has separable domains that function either to activate or suppress transcription. We now have identified a second WT1 binding sequence (5'-TCCTCCTCCTCCTCTCC-3') 3' to the transcription initiation site of the platelet-derived growth factor A-chain gene by DNase I footprinting and gel mobility shift assays. WT1 requires both 5' and 3' binding sites for transcriptional suppression; however, WT1 functions as a transcriptional activator when it binds to either the 5' or 3' site alone. This second WT1 binding sequence functions equally well as the previously identified 5'-GCGGGGGCG-3' sequence when analyzed in transient transfection assays. A core DNA sequence recognized by WT1 was defined by using related synthetic oligonucleotides. We also identified sequences similar to the WT1 binding site within the promoter regions of five other growth-related genes and demonstrated that each of these sequences also binds WT1 in gel mobility shift assays. These results thus identify a second WT1 binding site and suggest that additional growth-related genes may be transcriptionally influenced by WT1. PMID- 8415629 TI - Repair of 8-hydroxyguanine in DNA by mammalian N-methylpurine-DNA glycosylase. AB - 8-Hydroxyguanine is one of the major base lesions implicated in mutagenesis induced by ionizing radiation and radiomimetic agents. This lesion appears to be repaired by human cells via multiple pathways including the one that involves a base glycosylase. Mouse N-methylpurine-DNA glycosylase, responsible for the removal of N-alkylpurines in DNA that are induced by simple monofunctional alkylating agents, also releases 8-hydroxyguanine from DNA in vitro and in vivo in Escherichia coli. The human N-methylpurine-DNA glycosylase, with a lower preference for N-alkylguanine than the mouse protein, removes the oxidized base less efficiently than the mouse protein. The recombinant mammalian glycosylases can rescue E. coli lacking MutM (Fpg) protein, the DNA glycosylase that is primarily responsible for removing 8-hydroxyguanine from the bacterial DNA. PMID- 8415630 TI - Aging and resistance to oxidative damage in Caenorhabditis elegans. AB - The dauer larva state and the age-1 mutation, both of which extend life-span in Caenorhabditis elegans, were tested for hyperresistance to cellular damage that may be relevant to aging. The age-1 strain TJ401 displayed hyperresistance to oxidative stress relative to its parental strain. The activities of two enzymes that protect cells from oxidative damage, superoxide dismutase (SOD) and catalase, showed an age-dependent increase in mutant animals, which was not seen in the parental strain. These increases in activities paralleled the time course of the hyperresistance. The results are consistent with the age-1 gene product functioning as a negative regulator of SOD and catalase activities. In wild-type and age-1 dauer larvae, elevated levels of SOD activity, but not of catalase activity, were present when compared with young adults. The common increase in SOD activity prompted cloning the C. elegans Cu/Zn SOD gene. Its position on the physical map of the genome was in the region to which the age-1 gene has been genetically mapped, but it is unlikely that a mutation at the SOD locus confers the Age phenotype. Results support the free radical theory of aging by suggesting that the increased resistance to oxidative stress may be among the causes of increased longevity in both strain TJ401 and in the dauer larva. PMID- 8415631 TI - Cell proliferation, DNA repair, and p53 function are not required for programmed death of prostatic glandular cells induced by androgen ablation. AB - Androgen ablation induces programmed death of androgen-dependent prostatic glandular cells, resulting in fragmentation of their genomic DNA and the cells themselves into apoptotic bodies. Twenty percent of prostatic glandular cells undergo programmed death per day between day 2 and 5 after castration. During this same period, < 1% of prostatic glandular cells enter the S phase of the cell cycle, documenting that > 95% of these die in G0. During the programmed death of these G0 glandular cells, a futile DNA repair process is induced secondary to the DNA fragmentation. This futile DNA repair is not required, however, since inhibition of this process by > 90% with an appropriately timed hydroxy-urea dosing regimen had no effect upon the extent of the programmed death of these cells after castration. Likewise, p53 gene expression is not required since the same degree of cell death occurred in prostates and seminal vesicles after castration of wild-type and p53-deficient mice. PMID- 8415632 TI - On achieving better than 1-A accuracy in a simulation of a large protein: Streptomyces griseus protease A. AB - Computational methods are frequently used to simulate the properties of proteins. In these studies accuracy is clearly important, and the improvement of accuracy of protein simulation methodology is one of the major challenges in the application of theoretical methods, such as molecular dynamics, to structural studies of biological molecules. Much effort is being devoted to such improvements. Here, we present an analysis of a 187-ps molecular dynamics simulation of the serine protease Streptomyces griseus protease A in its crystal environment. The reproduction of the experimental structure is considerably better than has been achieved in earlier simulations--the root mean square deviation of the simulated structure from the x-ray structure being less than 1 A, a significant step toward the goal of simulating proteins to within experimental error. The use of a longer cutoff with truncation rather than a switching function, inclusion of all crystalline water and the counterions in the crystallization medium, and use of the consistent valence force field characterize the differences in this calculation. PMID- 8415633 TI - Structure of hydrated oligonucleotides studied by in situ scanning tunneling microscopy. AB - We have used the scanning tunneling microscope (STM) to image several synthetic oligonucleotides adsorbed onto a positively charged Au(111) electrode. The molecules were deposited and imaged in aqueous electrolyte under potential control, a procedure that eliminated the problem of the substrate artifacts that had limited some previous STM studies. Experiments were carried out with two types of single-stranded molecules (11 and 20 bases long) and three types of double-stranded molecules (20 and 61 base pairs and 31 bases with 25 bases paired and 6-base "sticky" ends). The molecules lie along symmetry directions on the reconstructed (23 x square root of 3) gold surface, and length measurements indicate that they adopt simple base-stacked structures. The base stacking distances are, within experimental uncertainty, equal to the 0.33 nm measured for polymeric aggregates of stacked purines by direct imaging in identical conditions. The images show features consistent with helical structures. Double helices have a major-groove periodicity that is consistent with a 36 degrees twist. The single helices appear to be more tightly twisted. A simple tunneling model of STM contrast generates good agreement between measured and calculated images. PMID- 8415634 TI - Origin of a novel allele in a mammalian hybrid zone. AB - The occurrence of rare or novel alleles has been documented in at least 23 different hybrid zones spanning vertebrate and invertebrate taxa. As most novel alleles either occur at high frequencies in hybrid populations or are exclusively restricted to hybrids, it has seemed probable that hybridization has a role in their origin; however, the molecular nature of these novel alleles and the mechanisms responsible for their origin remain obscure. We examined the complete coding sequences of six alleles of alcohol dehydrogenase in a mammalian hybrid zone between two species of pocket gophers (Geomys). One of these sequences encodes a novel electromorph that had been identified in earlier allozyme studies; this novel allele differs from one of the parental alleles by a single base substitution. This substitution generates an amino acid replacement that affects the net charge of the translated protein. This resultant charge change is congruent with the observed allozyme mobility patterns. Our data thus provide evidence for simple DNA substitution as a mechanism for the origin of this novel hybrid-zone allele. PMID- 8415635 TI - Selective loss of dopamine D3-type receptor mRNA expression in parietal and motor cortices of patients with chronic schizophrenia. AB - The expression of dopamine D3-subtype-receptor mRNA was analyzed in defined anatomic regions of brain obtained postmortem from patients with chronic schizophrenia and from controls. The specific amplification of D3-encoding cDNA by PCR allowed the identification of D3 mRNA expression in a wide variety of anatomic regions in both control brains and brains obtained from schizophrenic patients. However, in the parietal cortex (Brodmann areas 1, 2, 3, and 5) and motor cortex (Brodmann area 4), a selective loss of D3 mRNA expression was found in schizophrenia. A different D3 mRNA species was identified that appears to be widely expressed and that is still found in those regions of schizophrenic brains where D3 mRNA could not be detected. Compared with D3 mRNA this RNA is significantly less abundant, and at present its function (if any) is unclear. Many variables associated with either the course and/or the therapeutic management of the disease may account for the selective loss of D3 mRNA in the motor, somatosensory, and somatosensory association areas of schizophrenic brains. PMID- 8415636 TI - Translation of reovirus RNA species m1 can initiate at either of the first two in frame initiation codons. AB - The m1 species of reovirus RNA, which encodes the minor protein component mu 2, possesses two initiation codons, one "strong" according to Kozak rules and preceded by 13 residues (IC1), the other "weak" and located 49 codons downstream of the first (IC2). In reovirus-infected cells only IC2 is used, but initiation from IC1 can be activated, and efficiency of initiation from either initiation codon modulated over a wide range, by coupling unrelated sequences to either or both ends of m1 RNA. For example, when the M1 genome segment is cloned into the thymidine kinase gene of vaccinia virus in such a way that various "irrelevant" stretches of nucleotides comprising restriction endonuclease cleavage sites or promoter remnants are coupled to the 5' end of m1 RNA, translation of the resultant transcripts is also initiated at IC2, with frequencies controlled by the nature of the attached sequences. However, in rabbit reticulocyte lysates these same transcripts are translated from IC1 as well as from IC2, and transcripts in which m1 RNA is preceded by long sequences of encephalomyocarditis virus RNA (from the T7 polymerase-controlled pTM1 vector) are translated exclusively from IC1. By contrast, m1 RNA itself is translated only from IC2. It appears that the most important factor that controls the extent to which translation is initiated from IC1 and IC2 is their "availability," which is likely to be a function of the extent to which the regions on either side of them interact with each other (and also, to a lesser extent, with the 3' untranslated region) either directly or via interaction with host cell proteins. The effects described here are of considerable potential significance when genetic material is rearranged as a result of translocations, insertions, deletions, and amplifications--that is, when sequences that are normally separated are brought into apposition. PMID- 8415637 TI - Selective amplification of additional members of the ADP-ribosylation factor (ARF) family: cloning of additional human and Drosophila ARF-like genes. AB - The ADP-ribosylation factor (ARF) family is one of four subfamilies of the RAS superfamily of low molecular weight GTP-binding proteins (G proteins). Highly degenerate oligonucleotides encoding two conserved regions were used in a PCR reaction to amplify cDNAs encoding each of the known ARF proteins and eight additional cDNA fragments encoding previously unreported human members of the ARF family. Additional sequences were obtained from yeast or fly libraries by using this technique. These oligonucleotides specifically amplify members of the ARF family but not the structurally related G protein alpha subunits or members of the other three subfamilies of the RAS superfamily. Fragments obtained by PCR were used to obtain full-length sequences encoding highly homologous ARF-like (ARL) gene products from human and Drosophila melanogaster libraries, termed ARL2 and Ar184F, respectively. The encoded proteins are each 184 amino acids long and are 76% identical, with 40-45% identity to human ARF1 and Drosophila arf-like (arl) proteins. These genes appear to be generally expressed in human tissues and during Drosophila development. The purified human ARL2 protein differed in several biochemical properties from human ARF proteins, including the complete absence of ARF activity. Thus, the ARF family of low molecular weight GTP-binding proteins includes at least 15 distinct but structurally conserved members, including both the functionally conserved ARF proteins and the functionally disparate ARL proteins. The latter proteins currently comprise two distinct gene products in Drosophila (arl and ARL84F) and one in man (ARL2). PMID- 8415638 TI - Processing of a fusion protein by endoprotease in COS-1 cells for secretion of mature peptide by using a chimeric expression vector. AB - The subtilisin-related proprotein convertase furin is expressed in various mammalian tissues. Expecting that COS-1 cells have a furin-like endoprotease, we constructed a fusion expression vector for production of a recombinant foreign protein having no signal peptide or a protein in truncated form into secreted mature protein. A cDNA fragment encoding N-terminal procalcitonin (pro-CT) of human calcitonin precursor was inserted into the mammalian expression vector pME18S. We used PCR techniques to generate four kinds of cDNAs encoding the C terminus of the pro-CT with Arg residues at P4 (Arg-Xaa-Lys-Arg), P6 (Arg-Xaa-Xaa Xaa-Lys-Arg), or both (Arg-Xaa-Arg-Xaa-Lys-Arg), in addition to the Lys-Arg motif at the cleavage site, in order to determine the conditions for efficient processing in nonendocrine cells, such as COS-1 cells. The cDNA coding for the Fc fragment of human immunoglobulin G1 was fused in-frame to the cDNA encoding pro CT at its C terminus. Upon transfection of the chimeric plasmids into COS-1 cells, almost all of the fusion protein with the Arg residues at both P4 and P6 were processed into secreted Fc product, even without cotransfection of furin. These results indicate that COS-1 cells have a furin-like endoprotease and suggest that pro-CT, with the Arg residues at both P4 and P6, can be used as a carrier peptide for expression of a foreign protein having no signal peptide or a protein in truncated form in COS-1 cells. PMID- 8415639 TI - I kappa B alpha-mediated inhibition of v-Rel DNA binding requires direct interaction with the RXXRXRXXC Rel/kappa B DNA-binding motif. AB - Rel family proteins bind to kappa B DNA sites, form heterodimers with one another, and modulate expression of genes linked to kappa B motifs. I kappa B factors associate with Rel proteins, inhibit Rel DNA binding in vitro, and displace DNA from DNA-bound Rel complexes. We have investigated the mechanism by which the p40/I kappa B alpha inhibitor interfers with Rel DNA-binding activity. Here, we report that p40 contacts the RXXRXRXXC DNA-binding motif conserved in all Rel family proteins, in addition to associating with the nuclear localizing sequence. Competition assays with a Rel-derived peptide comprising the DNA binding region specifically alleviated p40-mediated inhibition of v-Rel DNA binding activity, whereas a covalently modified Rel peptide was inactive. Combined, these results indicate that I kappa B alpha interaction with the RXXRXRXXC motif is required for inhibition of v-Rel DNA binding and suggest that nuclear I kappa B factors may be critical for regulating transcription by Rel family proteins. PMID- 8415640 TI - ApcMin, a mutation in the murine Apc gene, predisposes to mammary carcinomas and focal alveolar hyperplasias. AB - ApcMin (Min, multiple intestinal neoplasia) is a point mutation in the murine homolog of the APC gene. Min/+ mice develop multiple intestinal adenomas, as do humans carrying germ-line mutations in APC. Female mice carrying Min are also prone to develop mammary tumors. Min/+ mammary glands are more sensitive to chemical carcinogenesis than are +/+ mammary glands. Transplantation of mammary cells from Min/+ or +/+ donors into +/+ hosts demonstrates that the propensity to develop mammary tumors is intrinsic to the Min/+ mammary cells. Long-term grafts of Min/+ mammary glands also gave rise to focal alveolar hyperplasias, indicating that the presence of the Min mutation also has a role in the development of these lesions. PMID- 8415641 TI - Precommitment of CD4+CD8+ thymocytes to either CD4 or CD8 lineages. AB - CD4+ and CD8+ mature T cells arise from CD4+CD8+ precursors in the thymus. During this process, cells expressing T-cell receptors (TCRs) reactive with self major histocompatibility complex (MHC) class I or II molecules are positively selected to the CD8 or CD4 lineage, respectively. It is controversial whether lineage commitment of CD4+CD8+ thymocytes is controlled directly by TCR specificity for MHC (instructional model) or, alternatively, by processes that operate independently of TCR specificity (stochastic model). We show here that CD4+CD8+ thymocytes bearing a MHC class I-restricted transgenic TCR can be subject to two alternative developmental fates. One population of CD4+CD8+ cells is positively selected by MHC class I molecules to the CD8 lineage as expected, whereas the other CD4+CD8+ population rearranges endogenous TCR genes and is positively selected by MHC class II molecules to the CD4 lineage. Blocking TCR-MHC class II interactions in vivo does not interfere with the generation of CD4+CD8+ cells expressing endogenous TCRs but does prevent their subsequent maturation to CD4+ cells. These data support a version of the stochastic model in which CD4+CD8+ thymocytes are precommitted to the CD4 or CD8 lineage independently of TCR specificity for MHC and prior to positive selection. PMID- 8415642 TI - Activation and differentiation requirements of primary T cells in vitro. AB - The progression of T cells from a quiescent or resting state to fully activated, proliferating cells is a crucial step in the initiation of an immune response. We have developed an in vitro system to study the requirements for triggering or hindering this pathway by using naive T cells derived from T-cell antigen receptor alpha beta transgenic animals and peptide-major histocompatibility (MHC) complexes coated on plates. Whereas previously stimulated T cells require only peptide-MHC complexes to produce interleukin 2 (IL-2), naive cells require at least one additional signal, which can be provided by either an anti-CD28 antibody or the protein kinase C stimulant phorbol 12-myristate 13-acetate. In contrast, the anti-CD28 antibody augments IL-2 production by primed T cells but is not required, and phorbol 12-myristate 13-acetate has no discernable effect. Thus we find that native T cells have significantly more stringent requirements for IL-2 production than primed cells and that this fits well with previous observations in other in vitro systems as well as in vivo models of autoimmunity. We also find that peptide-MHC complex stimulation of naive T cells, together with exogenous IL-2, is sufficient to convert these cells to primed T cells in vitro in 2 days, as assayed both by surface marker analysis and stimulation requirements. Taken together with the above results, this suggests that the activation of primary T cells requires at least two signals and that IL-2 produced by naive T cells in vivo may act in an autocrine fashion to allow them to proliferate and differentiate. PMID- 8415643 TI - Genes for E1, E2, and E3 small nucleolar RNAs. AB - We have found earlier three small nucleolar RNA (snoRNA) species, named E1, E2, and E3, that have unique nucleotide sequences and may participate in ribosome formation. The present report shows that there is a monophosphate at the 5' end of each of these three snoRNAs, suggesting that their 5' termini are formed by RNA processing. E1, E2, and E3 human genomic sequences were isolated. Apparently, the E2 and E3 loci are genes for the main E2 and E3 RNA species, based on their full homology, while the E1 locus is a gene for an E1 RNA sequence variant in HeLa cells. These loci do not have any of the intragenic or flanking sequences known to be functional in other genes. The E1 gene is located within the first intron of the gene for RCC1, a protein that regulates onset of mitosis. There is substantial sequence homology between the human E3 gene and flanking regions, and intron 8 and neighboring exons of the gene for mouse translation initiation factor 4AII. Injection of the human E1, E2, and E3 genes into Xenopus oocytes generated sequence-specific transcripts of the approximate sizes of the respective snoRNAs. We discuss why the available results are compatible with specific transcription and processing occurring in frog oocytes. PMID- 8415644 TI - RNase E activity is conferred by a single polypeptide: overexpression, purification, and properties of the ams/rne/hmp1 gene product. AB - Ribonuclease E, an enzyme that processes pre-5S rRNA from its precursor, is now believed to be the major endoribonuclease participating in mRNA turnover in Escherichia coli. The product of the ams/rne/hmp1 gene, which is required for RNase E activity, was overexpressed, purified to near homogeneity by electroelution from an SDS/polyacrylamide gel, and renatured. The purified polypeptide possesses nucleolytic activity in vitro with a specificity identical to that observed for crude RNase E preparations. In addition, both UV crosslinking and RNA-protein blotting unambiguously showed that the Ams/Rne/Hmp1 polypeptide has a high affinity for RNA. Our results demonstrate that RNase E activity is directly attributable to, and is an inherent property of, an RNA binding protein, the ams/rne/hmp1 gene product. PMID- 8415645 TI - SecA protein is required for translocation of a model precursor protein into inverted vesicles of Escherichia coli plasma membrane. AB - We have investigated whether the SecA protein is required for in vitro translocation of a model presecretory protein into inverted vesicles (INV) of the Escherichia coli plasma membrane. Contrary to previous reports, we found that urea-extracted INV that contained only the membrane-integral form of SecA were fully translocation active. Proteoliposomes that were reconstituted from a detergent extract of INV did contain a full complement of membrane-integral SecA but < 1% of SecY. These proteoliposomes were fully translocation active. However, immunodepletion of > 90% of the SecA from the detergent extract yielded proteoliposomes that were translocation inactive. Addition of purified SecA to the SecA-depleted proteoliposomes restored translocation. The amounts of SecA required to saturate translocation activity were equivalent to those present as membrane-integral SecA in INV. These data indicate that SecA is necessary for protein translocation, and reinforce our previous conclusion that SecY is not required. Contrary to previous reports, we find that membrane-integral SecA is not irreversibly inactivated by 6 M urea and that membrane-integral SecA and SecY do not form a stoichiometric protein complex in the membrane. PMID- 8415646 TI - A genotype of hepatitis D virus that occurs in northern South America. AB - Hepatitis D virus (HDV) is the cause of an unusually severe form of liver disease with distinct histologic features (morula cell) that occurs throughout northern South America and certain other areas of the world. Clinical studies of HDV disease worldwide indicate that there is, in fact, a wide variation in pathogenesis, and the reasons for these differences are presently unknown. One possible explanation is that factors associated with the viral genotype are determinants of HDV pathogenesis. In this study, nucleic acid sequences were determined for three different northern South American HDV isolates which were obtained from individuals with severe disease or a family history of severe disease, in areas that are hyperendemic for this disease pattern. The sequences of these three isolates are very similar to one another but only distantly related to other published HDV sequences. Comparison of the sequence of a semiconserved region from a total of 14 isolates indicates that there are at least three HDV genotypes. Most published HDV sequences, including those from North America, Europe, the Middle East, the South Pacific, and Asia, belong to a single genotype which may have some geographically based subtypes. A single Japanese isolate is the sole representative of a second HDV genotype. The South American sequences reported here constitute a third genotype. The association of a particular genotype with the severe form of type D hepatitis that occurs in northern South America supports the hypothesis that HDV genetic factors are important determinants in the pathogenesis of type D hepatitis. PMID- 8415647 TI - A chromatin folding model that incorporates linker variability generates fibers resembling the native structures. AB - The "30-nm" chromatin fibers, as observed in eukaryotic nuclei, are considered a discrete level in a hierarchy of DNA folding. At present, there is considerable debate as to how the nucleosomes and linker DNA are organized within chromatin fibers, and a number of models have been proposed, many of which are based on helical symmetry and imply specific contacts between nucleosomes. However, when observed in nuclei or after isolation, chromatin fibers show considerable structural irregularity. In the present study, chromatin folding is considered solely in terms of the known properties of the nucleosome-linker unit, taking into account the relative rotation between consecutive nucleosomes that results from the helical twist of DNA. Model building based on this premise, and with a constant length of linker DNA between consecutive nucleosomes, results in a family of fiber- and ribbon-like structures. When the linker length between nucleosomes is allowed to vary, as occurs in nature, fibers showing the types of irregularity observed in nuclei and in isolated chromatin are created. The potential application of the model in determining the three-dimensional organization of chromatin in which nucleosome positions are known is discussed. PMID- 8415648 TI - Altered expression of the cyclin D1 and retinoblastoma genes in human esophageal cancer. AB - We have examined DNA from four human esophageal carcinoma cell lines and 50 primary esophageal carcinomas obtained from China, Italy, and France for amplification of the cyclin D1 gene. We also examined 36 of these 50 carcinomas for expression of the cyclin D1 and retinoblastoma (RB) proteins by immunohistochemistry. We found a 3- to 10-fold amplification of the cyclin D1 gene in 16 of the 50 (32%) tumors and in two of the four cell lines. Cyclin D1 protein was overexpressed in 12 of 13 tumors and the two cell lines that showed gene amplification when compared to normal controls. Studies on RB protein expression indicated that 6 of the 36 (17%) tumor samples examined and one cell line did not show detectable expression of this protein. The tumors and cell lines that had cyclin D1 gene amplification and overexpression exhibited normal levels of expression of RB protein. By contrast, the tumors and cell line that did not appear to express the RB protein did not show amplification of the cyclin D1 gene and expressed only low levels of the cyclin D1 protein (P = 0.03). These results suggest that the inhibitory effect of RB on cell cycle progression can be abrogated during tumor development either by loss of expression of the RB gene or by increased expression of the cyclin D1 gene. PMID- 8415649 TI - Quaternary ligand binding to aromatic residues in the active-site gorge of acetylcholinesterase. AB - Binding sites of Torpedo acetylcholinesterase (EC 3.1.1.7) for quaternary ligands were investigated by x-ray crystallography and photoaffinity labeling. Crystal structures of complexes with ligands were determined at 2.8-A resolution. In a complex with edrophonium, and quaternary nitrogen of the ligand interacts with the indole of Trp-84, and its m-hydroxyl displays bifurcated hydrogen bonding to two members of the catalytic triad, Ser-200 and His-440. In a complex with tacrine, the acridine is stacked against the indole of Trp-84. The bisquaternary ligand decamethonium is oriented along the narrow gorge leading to the active site; one quaternary group is apposed to the indole of Trp-84 and the other to that of Trp-279, near the top of the gorge. The only major conformational difference between the three complexes is in the orientation of the phenyl ring of Phe-330. In the decamethonium complex it lies parallel to the surface of the gorge; in the other two complexes it is positioned to make contact with the bound ligand. This close interaction was confirmed by photoaffinity labelling by the photosensitive probe 3H-labeled p-(N,N-dimethylamino)benzenediazonium fluoroborate, which labeled, predominantly, Phe-330 within the active site. Labeling of Trp-279 was also observed. One mole of label is incorporated per mole of AcChoEase inactivated, indicating that labeling of Trp-279 and that of Phe-330 are mutually exclusive. The structural and chemical data, together, show the important role of aromatic groups as binding sites for quaternary ligands, and they provide complementary evidence assigning Trp-84 and Phe-330 to the "anionic" subsite of the active site and Trp-279 to the "peripheral" anionic site. PMID- 8415650 TI - Skeletal troponin C reduces contractile sensitivity to acidosis in cardiac myocytes from transgenic mice. AB - Depressed contractile function plays a primary role in the pathophysiology of acute myocardial ischemia. Intracellular acidification is an important factor underlying the inhibition of force production in the ischemic myocardium. The effect of acidosis to depress contractility is markedly greater in cardiac as compared to skeletal muscle; however, the molecular basis of this difference in sensitivity to acidosis is not clearly understood. In this report, we describe transgenic mice that express the fast skeletal isoform of troponin C (sTnC) in cardiac muscle. In permeabilized single cardiac myocytes the shift in the midpoint of the tension-pCa relationship (i.e., pCa50, where pCa is -log[Ca2+]) due to lowering pH from 7.00 to 6.20 was 1.27 +/- 0.03 (n = 7) pCa units in control cardiac TnC (cTnC) expressing myocytes and 0.96 +/- 0.04 (n = 11) pCa unit in transgenic cardiac myocytes (P < 0.001). The effect of pH to alter maximum Ca(2+)-activated tension was unchanged by TnC isoforms in these cardiac myocytes. In a reciprocal experiment, contractile sensitivity to acidosis was increased in fast skeletal muscle fibers following extraction of endogenous sTnC and reconstitution with purified cTnC in vitro. Our findings demonstrate that TnC plays an important role in determining the profound sensitivity of cardiac muscle to acidosis and identify cTnC as a target for therapeutic interventions designed to modify ischemia-induced myocardial contractile dysfunction. PMID- 8415651 TI - Site-directed mutations in a highly conserved region of Bacillus thuringiensis delta-endotoxin affect inhibition of short circuit current across Bombyx mori midguts. AB - Bacillus thuringiensis delta-endotoxins (Cry toxins) are insecticidal proteins of approximately 65 kDa in the proteolytically processed and active form. The structure of one of these toxins, CryIIIA, has been determined by Li et al. [Li, J., Carroll, J. & Ellar, D. J. (1991) Nature (London) 353, 815-821] and contains three domains. It is believed that other delta-endotoxins adopt similar three dimensional structure. Li et al. proposed that the first domain is the membrane pore-forming domain. Previous work from our laboratory has shown that the second domain is the receptor binding domain, but the function of the third domain is unclear. Site-directed mutagenesis was used to convert the "arginine face" of one of five highly conserved regions, QRYRVRIRYAS of CryIAa (residues 525-535), to selected other residues. This sequence corresponds to the beta-sheet 17 of CryIIIA in the third domain. Mutations in the second and third arginine positions resulted in structural alterations in the protein and were poorly expressed in Escherichia coli. Toxins from genes mutated to replace lysine for the first and fourth arginines were unaltered in expression and structure, as measured by trypsin activation, CD spectra, and receptor binding, but were substantially reduced in their insecticidal properties and inhibition of short circuit current across Bombyx mori midguts. It is proposed that this region plays a role in toxin function as an ion channel. PMID- 8415652 TI - A herpes simplex virus type 1 immediate-early gene product, IE63, regulates small nuclear ribonucleoprotein distribution. AB - Herpes simplex virus 1 (HSV-1), a nuclear replicating DNA virus, has 73 identified genes of which only 4 contain introns. For this reason the virus probably makes only minimal use of the cellular RNA-splicing machinery. Antigens associated with the small nuclear ribonucleoprotein particles (snRNPs) that are subunits of splicing complexes have been reported to redistribute in the nucleus and become concentrated into the intranuclear structures, the interchromatin granules, after HSV-1 infection [Martin, T. E., Barghusen, S. C., Leser, G. P. & Spear, P. G. (1987) J. Cell Biol. 105, 2069-2082]. We observe this snRNP redistribution upon HSV-1 infection, in which the widespread snRNP staining pattern changes to a restricted punctate distribution with a concomitant loss of coiled bodies in HSV-1-infected cells. We show here that expression of the immediate-early (IE) subset of HSV-1 genes is necessary and sufficient for snRNP redistribution. Using a series of HSV-1 mutants in different IE genes, we have established that specifically the product of the viral IE63 (ICP27) gene is essential for this effect, and transfection experiments revealed that IE63 expression alone can cause the snRNP redistribution. Further, we show that the IE63 gene product colocalizes with the redistributed snRNP in the nucleus. The snRNP redistribution caused by HSV-1 infection resembles the effect seen after inhibition of transcription in uninfected cells. In HSV-1-infected cells, however, the snRNP redistribution is under the control of viral IE gene products and occurs during active virus gene transcription. PMID- 8415653 TI - Increasing antigenic and genetic diversity of the V3 variable domain of the human immunodeficiency virus envelope protein in the course of the AIDS epidemic. AB - Population-wide variation in genomic RNA of human immunodeficiency virus type 1 (HIV-1) encompassing the V3 loop of the envelope protein was studied in serum samples of 74 newly infected individuals from three Dutch cohorts: 30 homosexual men, 32 drug users, and 12 hemophiliacs. During acute infection, HIV-1 RNA sequences present in serum are relatively homogeneous, which makes direct sequencing feasible. This offered an opportunity to study the infecting virus variants before mutations had accumulated in the new host. The sampling dates ranged from 1980 to 1991, thus spanning the entire AIDS epidemic in The Netherlands. The diversity in the sequenced region increased over time in both the homosexual and the drug-user risk groups. Furthermore, this increase was associated with an increase in antigenic variation, as witnessed by serum reactivity to a V3 peptide panel. Despite this diversification, some 1990 sequences still closely resembled the earliest 1980 sequence, making ancestral inferences problematic. No evidence was found of a change in the master sequence of the virus quasi-species over time. At the amino acid level, no risk-group associated variation was found, but at the nucleotide level, the drug-user and homosexual/hemophiliac sequences could be distinguished on the basis of a single silent nucleotide change in the sequence encoding the tip of the V3 loop. Hemophiliac sequences could not be distinguished from those of homosexuals. In spite of the large and increasing genetic variability, all sequences were more similar to the European/American HIV consensus sequence than to that of non Western strains. PMID- 8415654 TI - An interdigitated columnar mosaic of cytochrome oxidase, zinc, and neurotransmitter-related molecules in cat and monkey visual cortex. AB - There is considerable physiological evidence for the compartmentalization of mammalian visual cortex into functional columnar modules, representing features of visual information processing such as eye and orientation specificity. However, anatomical markers of visual cortical compartmentalization have been described only for primate visual cortex. In this report, we describe an interdigitated mosaic of four neuroactive molecules which demarcate two distinct columnar systems in the kitten visual cortex. Serotonin 1C receptors and synaptic zinc were found to demarcate columns within layer IV of kitten visual cortex, which were interdigitated with a second, patchy system characterized by increased levels of cytochrome oxidase and acetylcholinesterase. In primate visual cortex, as well as in the kitten, synaptic zinc was periodically distributed in a manner precisely complementary to cytochrome oxidase. These findings provide an anatomical framework on which unifying hypotheses of the functional organization of columnar systems in mammalian visual cortex can be built. PMID- 8415655 TI - Dominant expression of mRNA for prostaglandin D synthase in leptomeninges, choroid plexus, and oligodendrocytes of the adult rat brain. AB - Glutathione-independent prostaglandin D synthase [prostaglandin-H2 D-isomerase; (5Z,13E)-(15S)-9 alpha,11 alpha-epidioxy-15-hydroxyprosta-5,13-dienoate D isomerase, EC 5.3.99.2] is an enzyme responsible for biosynthesis of prostaglandin D2 in the central nervous system. In situ hybridization with antisense RNA for the enzyme indicated that mRNA for the enzyme was predominantly expressed in the leptomeninges, choroid plexus, and oligodendrocytes of the adult rat brain. The findings agree with those obtained by immunohistochemical staining with antibodies against the enzyme. It was further revealed that prostaglandin D synthase activity was considerably greater in the isolated leptomeninges (14.2 nmol per min per mg of protein) and choroid plexus (7.0 nmol per min per mg of protein) than the activity in the whole brain (2.0 nmol per min per mg of protein). These results, taken together, indicate that the enzyme is mainly synthesized and located in the leptomeninges, choroid plexus, and oligodendrocytes in the brain. PMID- 8415656 TI - Mice with reduced levels of p53 protein exhibit the testicular giant-cell degenerative syndrome. AB - Transgenic mice which carry hybrid p53 promoter-chloramphenicol acetyltransferase (CAT) transgenes were found to express CAT enzymatic activity predominantly in the testes. Endogenous levels of p53 mRNA and protein were lower than in the nontransgenic control mice. The various p53 promoter-CAT transgenic mice exhibited in their testes multinucleated giant cells, a degenerative syndrome resulting presumably from the inability of the tetraploid primary spermatocytes to complete meiotic division. The giant-cell degenerative syndrome was also observed in some genetic strains of homozygous p53 null mice. In view of the hypothesis that p53 plays a role in DNA repair mechanisms, it is tempting to speculate that the physiological function of p53 that is specifically expressed in the meiotic pachytene phase of spermatogenesis is to allow adequate time for the DNA reshuffling and repair events which occur at this phase to be properly completed. Primary spermatocytes which have reduced p53 levels are probably impaired with respect to DNA repair, thus leading to the development of genetically defective giant cells that do not mature. PMID- 8415657 TI - Regulation of bicarbonate-dependent ductular bile secretion assessed by lumenal micropuncture of isolated rodent intrahepatic bile ducts. AB - While intrahepatic bile duct epithelial cells secrete bile through transport of ions and water, the physiological mechanisms regulating ductular bile secretion are obscure, in part because of the lack of suitable experimental models. We report here the successful micropuncture of the lumen of isolated intrahepatic bile ducts and direct measurements of ductular ion secretion. Intact, polarized bile duct units (BDUs) were isolated from livers of normal rats by enzymatic digestion and microdissection. BDUs were cultured and mounted on a microscope in bicarbonate-containing buffer, and the lumens were microinjected with 2',7'-bis(2 carboxyethyl)-5-(and -6)carboxyfluorescein (BCECF)-dextran. Lumenal pH was measured by ratio imaging of BCECF fluorescence using digitized video fluorescent microscopy. After 36 hr in culture, the ends of BDUs sealed, forming closed compartments. After lumenal microinjection of BCECF-dextran, fluorescence was stable at the pH-insensitive wavelength, indicating no dye leakage. Serial changes in pH of extralumenal buffers containing pH-gradient collapsing ionophores allowed us to establish reliable standard curves relating fluorescence ratio to lumenal pH (r = 0.99; P < 0.001). By this approach, the basal pH inside the lumen of BDUs was 7.87 +/- 0.08 units (n = 9), 0.47 unit higher (P < 0.001) than the bathing buffer pH. Addition of 100 microM forskolin increased (P = 0.02) the lumenal pH from 7.78 +/- 0.06 to 7.97 +/- 0.06 units (n = 5); the forskolin effect was completely abolished by incubation of BDUs in HCO3-/CO2-free buffer. Moreover, forskolin caused a 50-fold increase in cAMP levels in BDUs. The observations are consistent with cAMP-dependent, active lumenal HCO3- secretion by BDUs. Furthermore, they demonstrate the suitability of the BDU model for studying regulatory and mechanistic aspects of ductular bile secretion. PMID- 8415658 TI - The human gastrin/cholecystokinin type B receptor gene: alternative splice donor site in exon 4 generates two variant mRNAs. AB - Gastrin and its carboxyl-terminal homolog cholecystokinin (CCK) exert a variety of biological actions in the brain and gastrointestinal tract that are mediated in part through one or more G protein-coupled receptors which exhibit similar affinity for both peptides. Genomic clones encoding a human gastrin/CCKB receptor were isolated by screening a human EMBL phage library with a partial-length DNA fragment which was based on the nucleotide sequence of the canine gastrin receptor. The gene contained a 1356-bp open reading frame consisting of five exons interrupted by 4 introns and was assigned to human chromosome 11p15.4. A region of exon 4, which encodes a portion of the putative third intracellular loop, appears to be alternatively spliced to yield two different mRNAs, one containing (452 amino acids; long isoform) and the other lacking (447 amino acids; short isoform) the pentapeptide sequence Gly-Gly-Ala-Gly-Pro. The two receptor isoforms may contribute to functional differences in gastrin- and CCK mediated signal transduction. PMID- 8415659 TI - The primary signal in the biological perception of temperature: Pd-catalyzed hydrogenation of membrane lipids stimulated the expression of the desA gene in Synechocystis PCC6803. AB - One of the well-characterized phenomena associated with the acclimation of organisms to changes in ambient temperature is the regulation of the molecular motion or "fluidity" of membrane lipids via changes in the extent of unsaturation of the fatty acids of membrane lipids. The enzymes responsible for this process when the temperature is decreased are the desaturases, the activities of which are enhanced at low temperature. To examine whether the change in the fluidity of membrane lipids is the first event that signals a change in temperature, we studied the effect of the Pd-catalyzed hydrogenation of membrane lipids on the expression of the desA gene, which is responsible for the desaturation of fatty acids of membrane lipids in the cyanobacterium Synechocystis PCC6803. The Pd catalyzed hydrogenation of plasma membrane lipids stimulated the expression of the desA gene. We also found that, for unexplained reasons, the hydrogenation was much more specific to a minor phospholipid, phosphatidylglycerol, than to members of other lipid classes. These results suggest that the organism perceives a decrease in the fluidity of plasma membrane lipids when it is exposed to a decrease in temperature. PMID- 8415660 TI - Three-dimensional structure of a membrane-containing virus. AB - The structure of Sindbis virus was determined by electron cryomicroscopy. The virion contains two icosahedral shells of viral-encoded proteins separated by a membrane bilayer of cellular origin. The three-dimensional structure of the ice embedded intact Sindbis virus, reconstructed from electron images, unambiguously shows that proteins in both shells are arranged with the same icosahedral lattice of triangulation number T = 4. These studies also provide structural evidence of contact between the glycoprotein and the nucleocapsid protein across the membrane bilayer. The structural organization of Sindbis virus has profound implications for the morphogenesis of the alphaviruses. The observed interactions confirm stoichiometric and specific protein associations that may be crucial for virion stability and predict a mechanism for assembly. PMID- 8415661 TI - Favored and suppressed patterns of hydrophobic and nonhydrophobic amino acids in protein sequences. AB - Hydrophobic amino acids of the group Leu, Ile, Val, Phe, and Met (LIVFM) are distributed in favored or suppressed patterns within protein sequences. The frequencies of all five-position combinations of [symbol: see text] = LIVFM and [symbol: see text] = non-LIVFM residues were analyzed in 48 proteins of known crystallographic structure. Some motifs were strongly preferred or suppressed; e.g., [symbol: see text] was favored (z = 3.5), while [symbol: see text] was suppressed (z = -3.4). In longer patterns, [symbol: see text] followed by [symbol: see text] and one [symbol: see text] was favored ([symbol: see text], z = 5.1), while conversion of the single hydrophobic residue to a pair was not ([symbol: see text], z = 0.8). Distributions of certain non-LIVFM amino acids around [symbol: see text] positions in strongly favored patterns were also favored or disfavored (Asp, Glu, Lys, Arg, Asn, Cys, Tyr, and Pro; for each magnitude of z > 2.0). While the strongly favored pattern [symbol: see text] was found in both alpha-helical and beta-strand sequences, it associated significantly with alpha-helices (z = 3.6 for the second-position alpha-helical phi and psi angles) but not with beta-strands (z = -1.1). Certain motifs of LIVFM and non-LIVFM residues might be selected if they lead efficiently to the local nucleations hypothesized to characterize molten globule intermediates in the folding of proteins. PMID- 8415662 TI - DNA polymerase delta from embryos of Drosophila melanogaster. AB - We have purified a DNA polymerase activity from 0- to 2-hr embryos of Drosophila melanogaster to near homogeneity. The purified enzyme consists of a single 120 kDa polypeptide, which contains polymerase and 3'-->5' exonuclease activities. Exonuclease activity is inhibited by deoxynucleoside triphosphates, suggesting that the polymerase and exonuclease activities are coupled. The polymerase is more active with poly(dA-dT) than with activated DNA or poly(dA)/oligo(dT) as template. It shows a low degree of processivity with poly(dA)/oligo(dT). The polymerase is sensitive to aphidicolin and carbonyldiphosphonate but resistant to N2-[p-(n-butyl)phenyl]-2-deoxyguanosine triphosphate, 2-[p-(n-butyl)anilino]-2 deoxyadenosine triphosphate, and dideoxythymidine triphosphate. The 120-kDa polypeptide can be distinguished from the large subunit of Drosophila DNA polymerase alpha on the basis of the peptides generated by partial cleavage with N-chlorosuccinimide and by its failure to react with a monoclonal antibody directed against the large subunit of DNA polymerase alpha. The DNA polymerase is inhibited by 200 mM NaCl and is unable to use poly(rA)/oligo(dT) as a template, thus differentiating it from DNA polymerase gamma. On the basis of these properties, we propose that the DNA polymerase that we have purified from 0- to 2 hr Drosophila melanogaster embryos is DNA polymerase delta. PMID- 8415663 TI - Functional characteristics of a cloned epithelial Na+/H+ exchanger (NHE3): resistance to amiloride and inhibition by protein kinase C. AB - We previously cloned an isoform Na+/H+ exchanger (NHE3), which was expressed only in intestine, kidney, and stomach. We show here the functional characteristics of NHE3 as a Na+/H+ exchanger by stably transfecting NHE3 cDNA into PS120 cells, a fibroblast cell line that lacks endogenous Na+/H+ exchangers. NHE3 was 39- and 160-fold more resistant to inhibition by amiloride and ethylisopropyl amiloride, respectively, than NHE1, the housekeeping Na+/H+ exchanger isoform. Although both exchangers were stimulated by serum, NHE3 was inhibited by phorbol 12-myristate 13-acetate (PMA), which stimulated NHE1. Mechanistically, serum and PMA stimulated NHE1 by an increase in the apparent affinity of the exchanger for intracellular H+. In contrast, serum stimulated and PMA inhibited NHE3 by a Vmax change. When NHE3 was stably expressed in Caco-2 cells, an intestinal epithelial cell line, NHE3 was functionally expressed in the apical membrane. Thus, NHE3 is a good candidate to be an epithelial brush border Na+/H+ exchanger. Furthermore, Na+/H+ exchangers can be rapidly regulated by mechanisms that change either the Vmax or the affinity for intracellular H+, depending on the Na+/H+ exchanger subtype. PMID- 8415664 TI - Fascin, an echinoid actin-bundling protein, is a homolog of the Drosophila singed gene product. AB - A cDNA for fascin, an actin-bundling protein in echinoderms, has been cloned, sequenced, and expressed. The predicted mass of the protein is approximately 55 kDa, similar to that observed for fascin purified from sea urchin eggs. Bacterially expressed fascin reacts with antibodies prepared against sea urchin egg fascin. Fascin has a strong sequence similarity to the singed gene (sn) product in Drosophila and has similarities with a 55-kDa human actin-bundling protein. No extensive similarities were found with other known actin binding/bundling proteins, indicating that this is a separate gene family. PMID- 8415665 TI - Expression and function of the interleukin 7 receptor in murine lymphocytes. AB - A monoclonal antibody, A7R34, that recognizes the high-affinity interleukin 7 receptor (IL-7Ra) and blocks the binding between IL-7 and IL-7Ra has been produced. Cell surface staining with A7R34 demonstrated that IL-7Ra is expressed in both B- and T-cell lineages. In the bone marrow, immature B-lineage cells that do not express surface IgM were IL-7Ra+. In the thymus, IL-7Ra was detected in CD4-8- T cells and also in CD4 or CD8 single-positive cells but not in CD4+8+ double-positive cells. In the peripheral lymphoid tissues, both CD4 and CD8 single-positive cells were the major cell types that express IL-7Ra. Addition of A7R34 to a long-term B-precursor-cell culture inhibited proliferation of the B lineage cells, indicating that IL-7 is an absolute requirement for in vitro B cell genesis. Consistent with this in vitro result, continuous injection of A7R34 into an adult mouse resulted in a decrease of B-precursor cells and also of thymocytes, whereas a considerable fraction of mature B and T cells in the peripheral tissues persisted over 2 weeks of the experiment. When A7R34 injection is started from day 14 of gestation, it is possible to produce mice that lack B cells. These results indicate that IL-7 is an essential molecule for generation of both B and T cells in murine bone marrow and thymus, respectively. Moreover, IL-7Ra would be the sole receptor system regulating these processes. PMID- 8415666 TI - Evidence that transporters associated with antigen processing translocate a major histocompatibility complex class I-binding peptide into the endoplasmic reticulum in an ATP-dependent manner. AB - We have investigated the role of the putative peptide transporters associated with antigen processing (TAP) by using a permeabilized-cell system. The main objective was to determine whether these molecules, which bear homology to the ATP-binding cassette family of transporters, translocate antigenic peptides across the endoplasmic reticulum membrane for assembly with major histocompatibility complex (MHC) class I molecules and beta 2-microglobulin light chain. The pore-forming toxin streptolysin O was used to generate permeabilized cells, and peptide translocation was determined by measuring the amount of added radiolabeled peptide bound to endogenous class I molecules. No radiolabeled peptide was associated with MHC class I glycoproteins from unpermeabilized cells. We found that efficient peptide binding to MHC class I molecules in permeabilized cells is both transporter dependent and ATP dependent. In antigen-processing mutant cells lacking a functional transporter, uptake occurs only through a less efficient transporter and ATP-independent pathway. In addition, short peptides (8 10 amino acids) known to bind MHC class I molecules compete efficiently with a radiolabeled peptide for TAP-dependent translocation, whereas longer peptides and a peptide derived from an endoplasmic reticulum signal sequence do not compete efficiently. This result indicates that the optimal substrates for TAP possess the characteristics of MHC-binding peptides. PMID- 8415667 TI - Protein hydration elucidated by molecular dynamics simulation. AB - Molecular dynamics (MD) simulation covering a wide range of hydration indicate that myoglobin is fully hydrated by 350 water molecules, in agreement with experiment. These waters, originally placed uniformly about the protein, form clusters that hydrate every charged group throughout the entire simulation. Some atoms in charged groups are hydrated by two water layers while 37% of the protein surface remains uncovered. The locations of the 350 waters are consistent with those of crystallographic waters resolved by x-ray and neutron diffraction. Hydration by 350 waters at 300 K stabilizes the conformation of carboxymyoglobin measured by x-ray diffraction throughout the entire protein, halves the rate of torsional transitions, and promotes alternative conformations for surface atoms. The glass transition observed experimentally in hydrated myoglobin near 220 K is also seen in the simulations and correlates with an increase in the number of dihedral angles undergoing transitions. The anharmonic protein motion above 220 K is enhanced by protein hydration. PMID- 8415668 TI - Replication-incompetent herpesvirus vector delivery of an interferon alpha gene inhibits human immunodeficiency virus replication in human monocytes. AB - Human monocytes and macrophages are nondividing cells that serve as a major reservoir for human immunodeficiency virus (HIV) at all stages of infection. To investigate viral-mediated gene delivery as a means of inhibiting HIV replication in human monocytes, a replication-incompetent herpes simplex virus vector was developed that expressed human interferon alpha. Monocytes infected with this herpes simplex virus vector and then challenged with HIV showed dramatically reduced cytopathic effects and HIV replication compared to control treated monocytes. Similar effects on HIV replication were observed if monocytes were first infected with HIV and then treated with the recombinant vectors. These results demonstrate that replication-incompetent herpes simplex virus gene delivery of interferon alpha directly to human monocytes can greatly decrease HIV replication and suggest that such a vector might deliver therapeutically important genes directly to sites of HIV infection. PMID- 8415669 TI - Amylin modulates beta-cell glucose sensing via effects on stimulus-secretion coupling. AB - The release of insulin from the pancreatic beta cell is dependent upon a complex interplay between stimulators and inhibitors. Recently, amylin, a peptide secreted by pancreatic beta cells, has been implicated in the development of type II (noninsulin dependent) diabetes through its modulation of the peripheral effects of insulin. However, the effect of amylin on insulin secretion from the beta cell has remained controversial. It is reported here that in single beta cells exhibiting normal glucose sensing, amylin causes membrane hyperpolarization, increases in net outward current, and reductions in insulin secretion. In contrast, in cells with abnormal glucose sensing (e.g., from db/db diabetic mice), amylin has no effect on electrical activity or secretion. Thus, amylin's effects on excitation-secretion coupling in the beta cell of the pancreas appear to be linked to the cell's capacity for normal glucose sensing. PMID- 8415670 TI - Epstein-Barr virus latent membrane protein 1 is essential for B-lymphocyte growth transformation. AB - The gene encoding latent-infection membrane protein 1 (LMP1) was specifically mutated in Epstein-Barr virus (EBV) recombinants by inserting a nonsense linker after codon 9 or codon 84 or into an intron 186 bp 3' to the latter insertion site. EBV recombinants with the LMP1 intron mutation were wild type for LMP1 expression and for growth transformation of primary B lymphocytes. In contrast, EBV recombinants with the mutations in the LMP1 open reading frame expressed N terminally truncated crossreactive proteins and could initiate or maintain primary B-lymphocyte transformation only when wild-type LMP1 was provided in trans by a coinfecting, transformation-defective EBV, P3HR-1. These data indicate that LMP1 is essential for EBV-mediated transformation of primary B lymphocytes, that the first 43 amino acids are critical for LMP1's function, and that codon 44 initiated LMP1 does not have a dominant negative effect on transformation. PMID- 8415671 TI - Salivary gland extracts from the deerfly contain a potent inhibitor of platelet aggregation. AB - Salivary gland extracts of the deerfly contain a potent inhibitor of platelet aggregation, which assists the insect in obtaining a blood meal. The extract prevents platelet aggregation induced by ADP, thrombin, and collagen and inhibits fibrinogen binding to the glycoprotein IIb/IIIa receptor on platelets. The active component in deerfly salivary gland extract appears to be a protein that is comparatively more potent than the disintegrins present in viper venoms. Isolation and characterization of this protein may provide different directions in therapeutics and studies of normal platelet physiology. PMID- 8415672 TI - Selection of CUG and AUG initiator codons for Drosophila E74A translation depends on downstream sequences. AB - Selection of a translation initiation site is thought to be determined by relative proximity to the 5' end and sequence context of a potential initiator codon. These guidelines seem insufficient to explain translation of the Drosophila E74A mRNA, whose 5' untranslated region is exceptionally long (1.8 kb) and contains many AUG triplets preceding the long open reading frame. In an effort to understand how the appropriate initiator codon is chosen, we have undertaken a study of E74A translation in transfected Drosophila cells. The results show that translation of the E74A protein utilizes at least three initiator codons: two minor forms of the protein are initiated at a CUG and an AUG, while the most abundant form initiates at a CUG. This main initiator CUG appears to be in a good context; however, it lies downstream of 17 AUG and 24 other CUG codons, several of which are also in good contexts. Unexpected results were obtained from sequence perturbations upstream and downstream of the main CUG initiator. Creating an AUG with a good context 72 bases 5' to the main CUG has only a modest inhibitory effect on initiation frequency at that CUG. Replacing sequences 44 bases 3' to the main CUG has an inhibitory effect on its use as an initiator as well as on the CUG 72 bases further upstream. These results indicate that factors other than context and relative proximity to the 5' end must be involved in initiator codon selection and may include elements such as secondary or tertiary structure of the RNA. PMID- 8415673 TI - Transport of glutathione diethyl ester into human cells. AB - Glutathione monoesters in which the carboxyl group of the glycine residue is esterified were previously found, in contrast to glutathione itself, to be effectively transported into various types of cells and to be converted intracellularly into glutathione. Glutathione monoesters are thus useful for prevention of oxidative stress, certain toxicities, and for treatment of glutathione deficiency. Glutathione diethyl ester is rapidly split to the glutathione monoethyl ester by mouse plasma glutathione diester alpha-esterase activity. Thus, as expected, glutathione mono- and diesters have similar effects on cellular glutathione levels in mice. However, human plasma lacks glutathione diester alpha-esterase; thus, it became of interest to compare the transport properties of glutathione mono- and diesters in human cells. We found that human cells (erythrocytes, peripheral blood mononuclear cells, fibroblasts, ovarian tumor cells, and purified T cells) transport glutathione diethyl ester much more effectively than the corresponding monoethyl (glycyl) ester. Human cells rapidly convert glutathione diethyl ester to the monoester, whose intracellular levels rise to levels that are significantly higher than levels found after application of the monoester to the cells. High levels of the monoester provide the cells with a means of producing glutathione over a period of time. We conclude that glutathione diethyl ester is highly effective as a delivery agent for glutathione monoester, and thus for glutathione, in human cells and therefore could serve to decrease oxidative stress and toxicity. Hamster (and certain other animals) also lack plasma glutathione diester alpha-esterase and therefore would be suitable animal models. Previously reported toxicity of certain glutathione ester preparations appears to reflect the presence of impurities rather than effects of the esters. PMID- 8415674 TI - Purification and characterization of recombinant G16 alpha from Sf9 cells: activation of purified phospholipase C isozymes by G-protein alpha subunits. AB - A cDNA encoding G16 alpha, the alpha subunit of a heterotrimeric guanine nucleotide-binding protein, was expressed in Sf9 cells using recombinant baculovirus. G16 alpha in membrane extracts of Sf9 cells activated phospholipase C-beta 1 (PLC-beta 1) in the presence of guanosine 5'-[gamma-thio]triphosphate; the system could not be activated by Al3+, Mg2+, and F-. The G16 alpha in the cytosolic fraction of Sf9 cells did not stimulate PLC-beta 1. Concurrent expression of the G-protein beta gamma subunit complex increased the amount of G16 alpha in Sf9 cell membranes. The guanosine 5'-[gamma-thio]triphosphate activated form of G16 alpha was purified from cholate extracts of membranes from cells expressing G16 alpha, and the G-protein beta 2 and gamma 2 subunits. G16 alpha activated PLC-beta 1, PLC-beta 2, and PLC-beta 3 in a manner essentially indistinguishable from that of Gq alpha. G16 alpha-mediated activation of PLC beta 1 and PLC-beta 3 greatly exceeded that of PLC-beta 2. G16 alpha did not activate PLC-gamma 1 or PLC-delta 1. Thus, two distantly related members of the Gq alpha family, Gq alpha and G16 alpha, have the same ability to activate the known isoforms of PLC-beta. PMID- 8415675 TI - A genetic method for defining DNA-binding domains: application to the nuclear receptor NGFI-B. AB - A method is described that allows for rapid and efficient generation of functional mutations in DNA-binding domains of proteins. The target DNA-binding domain is attached to the Gal4p transcriptional-activating domain and expressed in yeast. The binding site recognized by the target domain is placed upstream of a gene that produces a protein toxic to yeast cells, so that the chimeric protein activates its expression, providing a selection against DNA-binding domain function. The chimeric protein also activates expression of a gene necessary for histidine prototrophy, using a second DNA-binding domain included in the chimera (lexA), providing a selection against general activator mutations. Therefore, requiring growth in the absence of histidine focuses mutations to the target DNA binding domain. This method was applied to the DNA-binding domain of the nuclear receptor NGFI-B. Nearly all mutations obtained concurred with previous studies of NGFI-B and other nuclear receptors, verifying the functional validity of the mutational profile obtained. In addition, by coupling this selection scheme with the two-hybrid system [Chien, C.-t., Bartel, P. L., Sternglanz, R. & Fields, S. (1991) Proc. Natl. Acad. Sci. USA 88, 9578-9582], mutations that alter protein interaction domains could also be obtained. PMID- 8415676 TI - Protein phosphorylation inhibits production of Alzheimer amyloid beta/A4 peptide. AB - The major component of amyloid plaque cores and cerebrovascular amyloid deposits found in Alzheimer disease is the beta/A4 peptide, which is derived from the Alzheimer amyloid protein precursor (APP). Recent evidence suggests that abnormalities in beta/A4 peptide production or beta/A4 peptide aggregation may underlie cerebral amyloidosis. In the present study, treatment of cells with phorbol dibutyrate, which activates protein kinase C, and/or okadaic acid, which inhibits protein phosphatases 1 and 2A, reduced beta/A4 peptide production by 50 80%. These effects were observed with APP695 and APP751 expressed in stably transfected CHO cells, as well as with endogenous APP in human glioma (Hs 683) cells. Phorbol dibutyrate also decreased beta/A4 peptide production in cells expressing various mutant forms of APP associated with familial Alzheimer disease, one of which was reported to manifest greatly increased beta/A4 peptide production in cultured cells. Mastoparan and mastoparan X, compounds which can activate phospholipase C and hence protein kinase C, also decreased beta/A4 peptide production in CHO cells stably transfected with APP695. A model is presented in which decreases in beta/A4 peptide production can be achieved by accelerating the metabolism of APP through a nonamyloidgenic secretory pathway. PMID- 8415677 TI - Homologous recombination in the nuclear genome of Chlamydomonas reinhardtii. AB - Nuclear transformation of the unicellular green alga Chlamydomonas reinhardtii has thus far been characterized by integration of the introduced DNA into nonhomologous sites. In this study, the occurrence of homologous recombination events during transformation was investigated with the intent of developing strategies for gene targeting and gene disruption. Homologous recombination was monitored by using nonfunctional 5' and 3' deletion derivatives of the wild-type C. reinhardtii nit1 gene (encodes nitrate reductase) as selectable markers (p5' delta and p3' delta respectively) and the low reverting nit1-305 strain as the transformation recipient. After introduction of the DNA into the cell, intermolecular recombination between p5' delta and p3' delta occurs at a high frequency to restore a functional nit1 gene, indicating the presence of homologous recombination machinery in mitotic cells. Gene-targeting events at the nit1 locus were selected by restoring nit1-305 cells to prototrophy after transformation with only p5' delta and were confirmed by analysis of genomic DNA. By comparing the number of transformants obtained after transformation with p5' delta to the number obtained after transformation with a functional nit1 gene, the frequency of homologous-to-random integration events ranged between 1:1000 after glass bead-mediated transformation and 1:24 after bombardment with DNA coated tungsten microprojectiles. PMID- 8415678 TI - Adhesion of Bordetella pertussis to eukaryotic cells requires a time-dependent export and maturation of filamentous hemagglutinin. AB - Bordetella pertussis, the human pathogen of whooping cough, when grown at 22 degrees C is nonvirulent and unable to bind eukaryotic cells. In response to a temperature shift to 37 degrees C, the bacterium acquires the ability to bind eukaryotic cells in a time-dependent fashion. By studying in vitro the temperature-induced transition, from the nonvirulent to the virulent state, we found that binding to CHO cells is mediated by the Arg-Gly-Asp-containing domain of filamentous hemagglutinin (FHA), a protein with multiple binding specificities. This protein is synthesized as a 367-kDa polypeptide within 10 min after temperature shift, but requires 2 hr before it is detected on the bacterial cell surface and starts to bind CHO cells. Mutations affecting the cell surface export of FHA abolish bacterial adhesion to CHO cells, while mutations in the outer membrane protein pertactin strongly reduce binding. This suggests that multiple chaperon proteins are required for a correct function of FHA. Finally, several hours after maximum binding efficiency is achieved, the N-terminal 220 kDa portion of FHA that contains the binding regions is cleaved off, possibly to release the bacteria from the bound cells and facilitate spreading. The different forms of FHA may play different roles during bacterial infection. PMID- 8415679 TI - Mutations at U2555, a tRNA-protected base in 23S rRNA, affect translational fidelity. AB - A plasmid carrying a mutation in the highly conserved base U2555 in Escherichia coli 23S rRNA was isolated by selecting for suppression of the -1 frameshift mutation trpE91. U2555 is normally protected in chemical footprinting experiments by the aminoacyl residue of A-site-bound tRNA. Substitution of U2555 by adenine or guanine (but not by cytosine) increased readthrough of all three stop codons and +1 and -1 frameshifting. These effects on translational fidelity demonstrate the importance of U2555 for selection of the correct tRNA at the ribosomal A site. PMID- 8415680 TI - Bone morphogenetic protein 2 transiently enhances expression of a gene, Id (inhibitor of differentiation), encoding a helix-loop-helix molecule in osteoblast-like cells. AB - Bone morphogenetic protein 2 (BMP-2) is a potent inducer of differentiation of osteoblasts both in vivo and in vitro. We examined the action of BMP-2 on expression of a helix-loop-helix-type transcription factor, Id (inhibitor of differentiation), in osteoblast-like cells, as well as in osteoblast-enriched cells and possible precursor cells. To our surprise, BMP-2 enhanced Id gene expression in the cell types of osteoblastic lineage we examined. The maximal BMP 2 enhancement was observed within 24 hr in early proliferating cultures and the enhancement lasted up to 96 hr. The BMP-2 effect was not blocked by actinomycin D, while it was blocked by cycloheximide, suggesting that BMP-2 regulates Id gene expression at least in part via posttranscriptional events, which require protein synthesis. Other experiments indicated that BMP-2 did not further enhance Id mRNA levels promoted by dexamethasone, while BMP-2 did not resume the Id mRNA levels suppressed by 1,25-dihydroxyvitamin D3. Similar BMP-2 enhancement of Id message expression was also observed in osteoblast-enriched fetal rat calvaria cells as well as C3H10T1/2 cells. These results indicate that BMP-2 enhances expression of Id in early cultures of osteoblastic cells and suggest that enhancement of Id expression may somehow be involved in the promotion of differentiation by this cytokine in these osteoblastic cells and in their precursor cells. PMID- 8415681 TI - X-ray crystallographic identification of a protein-binding site for both all trans- and 9-cis-retinoic acid. AB - The elucidation of how a protein-binding site might specifically recognize both the all-trans and 9-cis isomers of retinoic acid is of particular interest because of the recently discovered binding specificities of the nuclear receptors for retinoic acid. Two families of nuclear receptors for retinoic acid have been described, which are designated RAR (for retinoic acid receptor) and RXR (for retinoid-X receptor). The RXR family of receptors is specific for 9-cis-retinoic acid, whereas the RAR-type receptor is activated by either 9-cis- or all-trans retinoic acid. During the x-ray structure determination of a secreted epididymal retinoic acid-binding protein, with and without retinoic acid, we observed an electron density for the bound all-trans-retinoic acid that indicates the protein bound all-trans form of the vitamin/hormone adopts a horseshoe-like conformation that resembles the structure of the 9-cis isomer of the ligand. We detail here the experiments that indicate the electron density is indeed due to all-trans retinoic acid and that protein can also bind the 9-cis isomer. This observation and the fact that the same protein also binds the synthetic retinoid (E)-4-[2 (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1- propenyl]-benzoic acid (TTNPB), a retinoic acid analog that activates RAR but does not activate RXR, suggest that the mechanism by which this protein recognizes both 9-cis- and all trans-retinoic acids may be analogous to the mechanism used by RAR. Three crystallographic structures of retinol-binding proteins have been described. In each of these structures the retinol binds with the isoprene tail fully extended. This report represents an x-ray crystallographic description of a protein-bound retinoid conformer that adopts a nonextended conformation, and we believe this observation is relevant to the ligand specificities described for the retinoic acid receptors. PMID- 8415682 TI - Specificity determinants of the P1 and P7 plasmid centromere analogs. AB - The cis-acting parS sites of P1 and P7 are similar in sequence and promote active partition of their respective plasmid prophages to daughter cells when the cognate Par proteins are supplied. Forty of the 94 relevant bases differ between the P1 and P7 parS sites, and the protein-site interactions show complete species specificity. A method was developed to predict which subset of the differing parS bases is responsible. When the four P1 bases thus identified were substituted into the P7 parS site, a complete switch to P1 specificity was observed. The P1 specific bases constitute two CG dinucleotide elements situated 66 bp apart. They lie within repeats of the TCGCCA sequence implicated in secondary contacts with the P1 ParB protein. The equivalent TC dinucleotides in the P7 site were found to be involved in P7 specificity. However, three other P7 bases can also contribute, including two in the heptamer repeats primarily responsible for ParB binding, and the P7-specific information shows some redundancy. The motifs containing the specificity dinucleotides and the primary ParB binding (heptamer) sites bear no obvious relationship of spacing or orientation to each other. For the ParB protein to contact both types of motif at the same time, the topology of the interaction must be complex. PMID- 8415683 TI - Functional analysis suggests unexpected role for conserved active-site residue in enzyme of known structure. PMID- 8415684 TI - The Epstein-Barr virus immortalizing protein EBNA-2 is targeted to DNA by a cellular enhancer-binding protein. AB - The Epstein-Barr virus nuclear antigen EBNA-2 is essential for Epstein-Barr virus induced immortalization of B cells. EBNA-2 is a transcriptional activator capable of modifying the expression of specific viral and cellular genes. However, the mechanism of EBNA-2 transactivation has been an enigma. We used a fractionated extract of CA46 lymphoblastoid cells and bacterially expressed EBNA-2 polypeptides to demonstrate that EBNA-2 is targeted to the Epstein-Barr virus latency C promoter (Cp) through interaction with a cellular DNA binding protein designated Cp binding factor 1 (CBF1). A glutathione S-transferase-EBNA-2 fusion protein containing aa 252-425 of EBNA-2 interacted with CBF1 to yield a slowly migrating complex in an electrophoretic mobility shift assay. Mutation of EBNA-2 aa 323 and 324, which lie within a highly conserved amino acid motif, abolished the interaction with CBF1. This same mutation also abolished the ability of EBNA 2 to activate the Cp in a cotransfection assay. The binding site for CBF1 was localized to residues -359 to -388 of the Cp by using an electrophoretic mobility shift assay and DNase I footprinting. Introduction of multiple copies of the CBF1 binding site upstream of a minimal heterologous promoter conferred EBNA-2 responsiveness on that promoter. Mutation of a core sequence CNGTGGGAA abolished CBF1 binding, and the mutated sequence was unable to mediate EBNA-2 transactivation. The CBF1 core sequence also occurs in other EBNA-2-responsive promoters suggesting that CBF1 may mediate EBNA-2 transactivation of both cellular and viral targets. PMID- 8415685 TI - The boundaries of partially edited transcripts are not conserved in kinetoplastids: implications for the guide RNA model of editing. AB - We have studied partially edited molecules for the cytochrome-c oxidase subunit III (COIII) transcript from two species of the insect trypanosome Herpetomonas. We found unexpected patterns of editing, in which editing does not proceed strictly 3' to 5', in 24 of 61 partially edited clones. A comparison of the partially edited molecules between the two kinetoplastid species revealed an 8- to 10-nt shift in precisely defined editing boundaries, sites at which editing pauses before binding of the next guide RNA after formation of a stable duplex between a guide RNA and mRNA. This suggests that the region of base pairing between individual guide RNAs and the COIII transcript is not strictly conserved in kinetoplastids, implying gradual evolution of the editing process. PMID- 8415686 TI - Raf-1 protein kinase activates the NF-kappa B transcription factor by dissociating the cytoplasmic NF-kappa B-I kappa B complex. AB - Addition of mitogenic growth factors to quiescent cells triggers complex signal transduction cascades that result in the reprogramming of gene expression and entry into the cell cycle. We have found that an oncogenic variant of the c-Raf-1 protein kinase stimulated the expression of promoters containing NF-kappa B binding sites. In situ immunofluorescence analysis revealed elevated nuclear levels of the p65 subunit of NF-kappa B in v-raf-transformed NIH 3T3 cells. Incubation of HeLa cell cytoplasmic extracts with a purified recombinant glutathione S-transferase-raf fusion protein in the presence of ATP released active NF-kappa B that could be detected by electrophoretic gel mobility shift assay. Coincubation of purified recombinant I kappa B and glutathione S transferase-raf in the presence of ATP resulted in the phosphorylation of I kappa B. Coexpression of GAL4 (activation domain)-I kappa B and GAL4 (DNA-binding domain)-raf fusion proteins in yeast resulted in stimulation of a GAL4-responsive reporter gene, indicating that I kappa B and Raf interact physically in vivo. These results indicate that the Raf-1 kinase functions in signal transduction in part by activating the NF-kappa B transcription factor by phosphorylating I kappa B in the cytoplasmic I kappa B-NF-kappa B complex to release active NF-kappa B. PMID- 8415687 TI - Modulation of cell proliferation and gene expression by a p53-estrogen receptor hybrid protein. AB - We report that p53her, a chimeric protein consisting of the complete human wild type p53 and the human estrogen receptor hormone-binding domain, strongly suppresses proliferation and induces characteristic morphological changes in Saos 2 human osteosarcoma cells when induced by 17 beta-estradiol. In contrast, p53her constitutively transactivates a p53-responsive promoter in transfection assays, so that transactivation is not regulated by estradiol. However, coexpression of p53her and oncoprotein MDM-2, which associates with and presumably inactivates p53, results in suppression of p53her-mediated transactivation in the absence, but not the presence, of estradiol. Similarly, p53her induces expression of an endogenous MDM-2 transcript only in the presence of estradiol. These results suggest a correlation between the growth suppressor function of p53her and release of a transactivation block mediated by MDM-2. PMID- 8415688 TI - Vitamin D-influenced gene expression via a ligand-independent, receptor-DNA complex intermediate. AB - A lingering question regarding the regulation of target gene expression by 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3] has been the delineation of vitamin D receptor (VDR)-DNA binding and transactivation. This report confirms that initial VDR-DNA interaction occurs in a ligand-independent fashion. An electrophoretic mobility shift analysis demonstrated that VDR, derived from extracts of the small intestines of vitamin D-deficient rats, is capable of binding a vitamin D response element (DRE). Additional mobility-shift studies using either porcine derived VDR or recombinant rat VDR from insect cells revealed DRE-binding capability in the absence of 1,25-(OH)2D3. The reactions were performed in various salt environments, with the maximum of porcine VDR-DRE and rat VDR-DRE binding detected at 100 mM and 150 mM KCl, respectively. The addition of 1,25 (OH)2D3 to an identical set of reaction mixtures resulted in increased DRE binding with greater affinities exhibited by both VDR types. These two phenomena were confirmed upon examination of an elution profile of VDR bound to DRE-linked Sepharose. When a linear KCl gradient was used for elution without the addition of 1,25-(OH)2D3, the peak of VDR was 205 mM KCl; the presence of exogenous hormone shifted the maximum VDR elution to a position corresponding to 265 mM KCl. Based on these data and previous reports on VDR-mediated transactivation, we propose a model for 1,25-(OH)2D3-influenced target gene expression. PMID- 8415689 TI - Replacement of serine-871 of hamster 3-hydroxy-3-methylglutaryl-CoA reductase prevents phosphorylation by AMP-activated kinase and blocks inhibition of sterol synthesis induced by ATP depletion. AB - An AMP-activated protein kinase has been reported to phosphorylate rodent 3 hydroxy-3-methylglutaryl-coenzyme A reductase [HMG-CoA reductase; (S)-mevalonate: NAD+ oxidoreductase (CoA-acylating), EC 1.1.1.88] at Ser-871, thereby lowering its catalytic activity [Clarke, P. R. & Hardie, D. G. (1990) EMBO J. 9, 2439 2446]. To explore the physiologic role of this reaction, we prepared a cDNA encoding a mutant form of hamster HMG-CoA reductase with alanine substituted for serine at residue 871. When overexpressed in transfected cells, the wild-type enzyme, but not the Ser-871 to Ala mutant, was labeled with [32P]phosphate, confirming Ser-871 as the site of phosphorylation. The wild-type enzyme, but not the mutant enzyme, showed reduced activity when the cells were harvested with the phosphatase inhibitor KF, confirming phosphorylation as a mechanism for inactivation within the cell. Despite the lack of phosphorylation, the posttranscriptional feedback regulation of the mutant enzyme was normal, as indicated by reduced activity when cells were incubated with mevalonate, 25 hydroxycholesterol, or low density lipoprotein. Moreover, the mutant enzyme showed a normal acceleration of degradation when the transfected cells were incubated with sterols. Cells expressing the wild-type enzyme showed a decreased incorporation of [14C]pyruvate into sterols when ATP was depleted by incubation with 2-deoxy-D-glucose. No such reduction was seen in cells expressing the Ser 871 to Ala mutant enzyme. We conclude that the AMP-activated protein kinase does not play a role in end-product feedback regulation of HMG-CoA reductase, but rather it comes into play when cellular ATP levels are depleted, thereby lowering the rate of cholesterol synthesis and preserving the energy stores of the cell. PMID- 8415690 TI - Endothelin receptors in rat liver: lipocytes as a contractile target for endothelin 1. AB - The endothelins (ETs) form a group of three vasoactive peptides (ET-1, ET-2, and ET-3) for which two types of cellular receptors have been identified, types A and B ET receptors (ETA and ETB receptors, respectively). To address possible targets for ETs within the liver, we isolated the four principal liver cell populations and placed them in short-term primary culture. By ligand-binding assay and mRNA levels, expression of ET receptors was greatest on hepatic lipocytes (Ito cells or fat-storing cells), which are perisinusoidal cells exhibiting features of smooth muscle cells. Moreover, lipocytes expressed both ETA and ETB receptors. The mRNA for ETB receptor, but not for ETA receptor, was detectable in sinusoidal endothelial cells and Kupffer cells; neither mRNA was detectable in hepatocytes. Both ET-1 and ET-3 elicited contraction of activated lipocytes cultured on collagen lattices; the EC50 value for ET-1 was 3 +/- 1 nM and for ET-3 was 17 +/- 12 nM. In cell isolates from injured liver (after administration of carbon tetrachloride), expression of ET receptors was unchanged. However, mRNA for ET-1 was significantly increased in activated lipocytes, suggesting an autocrine loop for the initiation of lipocyte contraction. The findings imply that ET-1 may play a role in regulating sinusoidal perfusion through its effect on lipocytes, particularly in injury states. PMID- 8415691 TI - Chromosome walking on the TCL1 locus involved in T-cell neoplasia. AB - The TCL1 locus on chromosome 14 band q32.1 is frequently involved in the chromosomal translocations and inversions with the T-cell receptor genes observed in several T-cell tumors, including T-prolymphocytic leukemias, acute and chronic leukemias associated with the immunodeficiency syndrome ataxia-telangiectasia, and adult T-cell leukemia. All breakpoints cloned in this area have been mapped to 14q32.1, an area distant approximately 10,000 kb from the immunoglobulin heavy chain gene locus on chromosome 14q band 32.3. Except for two cases of inversion, no physical linkage of the cloned breakpoints has been reported, nor has a gene been identified in this region. Taking advantage of chromosome-walking techniques and of the P1 phage, we cloned and characterized 450 kb of the germ-line TCL1 locus, starting from the breakpoints of two independent T-cell leukemias. We show that all molecular rearrangements characterized so far map to these clones, indicating not only that this region is the target of chromosomal rearrangements occurring in this area but also that both inversion and translocations occur within a 300-kb region in the T-cell leukemias. In the attempt to identify a candidate oncogene responsible for the malignant transformation, a CpG island centromeric to the inversions and to the translocations has been identified. Two probes near the CpG island have detected sequences conserved among species, as well as two transcripts in the K562 human erythroleukemia cell line. On the basis of these data, a model of activation of the putative TCL1 oncogene is suggested. PMID- 8415692 TI - Cloning, sequence determination, and regulation of the ribonucleotide reductase subunits from Plasmodium falciparum: a target for antimalarial therapy. AB - Malaria remains a leading cause of morbidity and mortality worldwide, accounting for more than one million deaths annually. We have focused on the reduction of ribonucleotides to 2'-deoxyribonucleotides, catalyzed by ribonucleotide reductase, which represents the rate-determining step in DNA replication as a target for antimalarial agents. We report the full-length DNA sequence corresponding to the large (PfR1) and small (PfR2) subunits of Plasmodium falciparum ribonucleotide reductase. The small subunit (PfR2) contains the major catalytic motif consisting of a tyrosyl radical and a dinuclear Fe site. Whereas PfR2 shares 59% amino acid identity with human R2, a striking sequence divergence between human R2 and PfR2 at the C terminus may provide a selective target for inhibition of the malarial enzyme. A synthetic oligopeptide corresponding to the C-terminal 7 residues of PfR2 inhibits mammalian ribonucleotide reductase at concentrations approximately 10-fold higher than that predicted to inhibit malarial R2. The gene encoding the large subunit (PfR1) contains a single intron. The cysteines thought to be involved in the reduction mechanism are conserved. In contrast to mammalian ribonucleotide reductase, the genes for PfR1 and PfR2 are located on the same chromosome and the accumulation of mRNAs for the two subunits follow different temporal patterns during the cell cycle. PMID- 8415693 TI - Endothelial-cell heme uptake from heme proteins: induction of sensitization and desensitization to oxidant damage. AB - Iron-derived reactive oxygen species are implicated in the pathogenesis of various vascular disorders including atherosclerosis, vasculitis, and reperfusion injury. The present studies examine whether heme, when liganded to physiologically relevant proteins as in hemoglobin, can provide potentially damaging iron to intact endothelium. We demonstrate that reduced ferrohemoglobin, while relatively innocuous to cultured endothelial cells, when oxidized to ferrihemoglobin (methemoglobin), greatly amplifies oxidant (H2O2)-mediated endothelial-cell injury. Drawing upon our previous observation that free heme similarly primes endothelium for oxidant damage, we posited that methemoglobin, but not ferrohemoglobin, releases its hemes that can then be incorporated into endothelial cells. In support, cultured endothelial cells exposed to methemoglobin--in contrast to exposure to ferrohemoglobin, cytochrome c, or metmyoglobin--rapidly increased their heme oxygenase mRNA and enzyme activity, thereby supporting heme uptake; ferritin production was also markedly increased after such exposure, thus attesting to eventual incorporation of Fe. These cellular methemoglobin effects were inhibited by the heme-scavenging protein hemopexin and by haptoglobin or cyanide, agents that strengthen the liganding between heme and globin. If the endothelium is exposed to methemoglobin for a more prolonged period (16 hr), it accumulates large amounts of ferritin; concomitantly, and presumably associated with iron sequestration by this protein, the endothelium converts from hypersusceptible to hyperresistant to oxidative damage. We conclude that when oxidation of hemoglobin facilitates release of its heme groups, catalytically active iron is provided to neighboring tissue environments. The effect of this relinquished heme on the vasculature is determined both by extracellular factors--i.e., plasma proteins, such as haptoglobin and hemopexin--as well as intracellular factors, including heme oxygenase and ferritin. Acutely, if both extra- and intracellular defenses are overwhelmed, cellular toxicity arises; chronically, when ferritin is induced, resistance to oxidative injury may supervene. PMID- 8415694 TI - A site required for termination of packaging of the phage lambda chromosome. AB - Lambda chromosomes are cut and packaged from concatemeric DNA by phage enzyme terminase. Terminase initiates DNA packaging by binding at a site called cosB and introducing staggered nicks at an adjacent site, cosN, to generate the left cohesive end of the DNA molecule to be packaged. After DNA packaging terminase recognizes and cuts the terminal cosN, an event that does not require a wild-type cosB. In this work a site, called cosQ, has been identified that is required for termination of DNA packaging. cosQ, defined by mutations in a sequence called R4, is located approximately 30 bp upstream from cosN. The order of sites is cosQ cosN-cosB. Helper packaging of repressed, tandem prophage chromosomes demonstrated that a cosQ point mutation affects DNA packaging only when placed at the terminal cos site, whereas cosB mutations only affect packaging initiation. In vitro packaging studies confirmed that cosQ mutations do not affect packaging initiation. In vivo studies indicated that cosQ mutations do not affect cutting of initial cos sites but do cause a defect in packaging termination. cosQ mutants accumulated expanded phage heads, indicating that cosQ mutations affect a step that occurs after packaging of a substantial length of phage DNA. These results show that cosQ mutations define a site required for use of cos sites present at the ends of lambda chromosomes undergoing packaging. Available evidence suggests that other viruses, including phages T3 and T7 and the herpesviruses, may ultimately prove to use cosQ-like sites for packaging termination. PMID- 8415695 TI - Transformation of Tetrahymena thermophila by microinjection of a foreign gene. AB - Tetrahymena thermophila has been transformed to paromomycin-resistant phenotypes by microinjection of an aminoglycoside 3'-phosphotransferase (neo) gene under the control of the T. thermophila histone H4-I promoter. This chimeric neo gene, by itself or on a vector containing a rRNA-encoding DNA (rDNA) origin of replication, transforms T. thermophila. In cells transformed with the rDNA origin vector, the neo gene is usually found integrated into the endogenous rDNA molecules and is present in high copy number. In transformants obtained by microinjecting only the linear chimeric gene, the neo gene is found to have replaced the histone H4-I gene or is found integrated into the 5' flanking region of the H4-I gene. The relative transcript levels of the neo gene in T. thermophila transformed by the linear chimeric gene are much higher than in cells transformed with the vector. The neo gene provides an effective selectable marker for transformation of T. thermophila. PMID- 8415696 TI - glh-1, a germ-line putative RNA helicase from Caenorhabditis, has four zinc fingers. AB - We have cloned a family of putative RNA helicases from the free-living nematode Caenorhabditis elegans. One of these, a cDNA that we call glh-1, most closely matches in sequence and expression the previously described germ-line helicases PL10 from mouse and vasa from Drosophila. The amino terminus of the predicted protein of glh-1 contains a set of glycine-rich repeats similar in location and sequence to those in the predicted vasa protein. However, unlike all other putative RNA helicases, glh-1 also contains four retroviral-type zinc fingers. The RNA expression pattern of this Caenorhabditis helicase correlates with the presence of germ-line tissue in the parasitic nematode Ascaris lumbricoides var. suum and with the presence of germ cells in wild type and several germ-line mutants of Caenorhabditis. In the germ-line mutants glp-4 and glp-1, additional larger species of glh-1 RNA exist, which correspond to different adenylylated forms of the glh-1 transcript; these may be specified by motifs in the 3' untranslated region of glh-1 that are similar to adenylylation control elements and nos response elements. PMID- 8415697 TI - Regional expression and chromosomal localization of the delta opiate receptor gene. AB - The delta opiate receptor gene has been cloned from the mouse neuroblastoma-rat glioma hybrid cell NG108-15. The clone that we isolated is apparently identical to that reported by Evans et al. [Evans, C. J., Keith, D. E., Jr., Morrison, H., Magendzo, K. & Edwards, R. H. (1992) Science 258, 1952-1955] and essentially identical with that of Kieffer et al. [Kieffer, B. L., Befort, K., Gaveriaux Ruff, C. & Hirth, C. G. (1992) Proc. Natl. Acad. Sci. USA 89, 12048-12052]. We have found full-length transcripts of the gene in mouse brain but in no other tissues examined. Within the brain the gene is expressed at low levels in many regions but transcripts are found in particularly large amounts in the anterior pituitary and pineal glands. Since these tissues are located outside the blood brain barrier, opioid peptides easily can reach receptors in these areas from the blood. The gene, which is present as a single copy, has been mapped to the distal region of mouse Chromosome 4. PMID- 8415698 TI - Maize mitochondrial manganese superoxide dismutases are encoded by a differentially expressed multigene family. AB - We have isolated maize cDNAs encoding three manganese-containing superoxide dismutases (MnSODs) distinct from the one previously reported. Molecular analyses indicate that multiple MnSOD transcripts are encoded by different, though similar, genes in the maize genome. A single MnSOD gene has been reported in all other organisms examined to date. The deduced amino acid sequences show that these maize MnSOD proteins have a mitochondrial transit peptide and that the first 9 amino acids (matrix-targeting sequence) in the transit peptide are conserved. This suggests that all the maize MnSOD proteins are mitochondria associated isozymes. RNA blot analysis demonstrated that each member of the maize MnSOD multigene family is both spatially and developmentally regulated. One gene, Sod3.3, was predominantly expressed in the embryo late in embryogenesis. Patterns of increased Mn-SOD transcript accumulation are shown to be associated with increased mitochondrial activity during plant growth and development. The influence of mitochondrial metabolism on the expression of the nuclear MnSOD genes is discussed. PMID- 8415699 TI - Multiple nucleosome positioning with unique rotational setting for the Saccharomyces cerevisiae 5S rRNA gene in vitro and in vivo. AB - A simple no-background assay was developed for high-resolution in vivo analysis of yeast chromatin. When applied to Saccharomyces cerevisiae 5S rRNA genes (5S rDNA), this analysis shows that nucleosomes completely cover this chromosomal region, occupying alternative positions characterized by a unique helical phase. This supports the notion that sequence-intrinsic rotational signals are the major determinant of nucleosome localization. Nucleosomal core particles reconstituted in vitro occupy the same positions and have the same helically phased distribution observed in vivo, as determined by mapping of exonuclease III resistant borders, mapping by restriction cleavages, and by DNase I and hydroxyl radical digestion patterns. PMID- 8415700 TI - Recognition of specific DNA sequences by the c-myb protooncogene product: role of three repeat units in the DNA-binding domain. AB - The DNA-binding domain of c-Myb consists of three homologous tandem repeats of 52 amino acids. The structure of the third (C-terminal) repeat obtained by NMR analysis has a conformation related to the helix-turn-helix motif. To identify the role of each repeat in the sequence recognition of DNA, we analyzed specific interactions between c-Myb and DNA by measuring binding affinities for systematic mutants of Myb-binding DNA sites and various truncated c-Myb mutants. We found that specific interactions are localized unevenly in the AACTGAC region in the consensus binding site of c-Myb: The first adenine, third cytosine, and fifth guanine are involved in very specific interactions, in which any base substitutions reduce the binding affinity by > 500-fold. On the other hand, the interaction at the second adenine is less specific, with the affinity reduction in the range of 6- to 15-fold. The seventh cytosine involves a rather peculiar interaction, in which only guanine substitution abolishes the specific binding. The binding analyses, together with the chemical protection analyses, showed that the c-Myb fragment containing the second and third repeats covers the AACTGAC region from the major groove of DNA in such an orientation that the third repeat covers the core AAC sequence. These results suggest that the third repeat recognizes the core AAC sequence very specifically, whereas the second repeat recognizes the GAC sequence in a more redundant manner. The first (N-terminal) repeat, which covers the major groove of DNA only partially, is not significant in the sequence recognition, but it contributes to increase the stability of the Myb-DNA complex. The presence of an N-terminal acidic region upstream of the first repeat, which is important for the activation of c-myb protooncogene, was found to reduce the binding affinity by interfering with the first repeat in binding to DNA. PMID- 8415701 TI - Synergism in replication and translation of messenger RNA in a cell-free system. AB - Combination of the Q beta replicase reaction with the Escherichia coli cell-free translation system markedly enhances replication of a recombinant RQ-DHFR RNA consisting of the dihydrofolate reductase (DHFR) mRNA sequence inserted into RQ135(-1) RNA, an efficient naturally occurring Q beta replicase template. The enhancement is associated with a replication asymmetry previously described for the replication of Q beta phage RNA in vivo; the sense (+)-strands are produced in large excess over the antisense (-)-strands. This, in turn, results in increased synthesis of the functionally active DHFR. These effects are not observed when DHFR mRNAs or RQ135(-1) RNAs are used as templates, if the translation system is not complete, or if it is inhibited by puromycin. The coupled replication-translation of nonviral mRNA recombinants can serve as a useful model for studying the fundamental aspects of virus amplification and can be implemented for large-scale protein synthesis in vitro. PMID- 8415702 TI - A hypothesis for the HLA-B27 immune dysregulation in spondyloarthropathy: contributions from enteric organisms, B27 structure, peptides bound by B27, and convergent evolution. AB - Several human rheumatic diseases occur predominantly in persons who carry the histocompatibility (HLA) class I allele B27. They have also been related to Gram negative enteric microorganisms. In addition, the recent recovery of peptides bound to B27 has allowed an understanding of the structural requirements for their binding. Using the accumulated data base of protein sequences, we have tested a series of hypotheses. First, we have asked whether the primary amino acid sequence of the hypervariable regions of HLA-B27 shares short sequences with the proteins of Gram-negative enteric bacteria. The data demonstrate that, unique among the HLA-B molecules, the hypervariable regions of HLA-B27 unexpectedly share short peptide sequences with proteins from these bacteria. Second, we have asked whether the enteric proteins tend to satisfy the structural requirements for peptide binding to B27 in those regions of the sequence shared with B27. This hypothesis also tends to be true, especially in an allelically variable part of the B27 sequence which is predicted to bind B27 if it were to be presented as a free peptide. We conclude that HLA-B27 and enteric Gram-negative bacteria have undergone a previously unappreciated form of convergent evolution which may be important in the process leading to these rheumatic diseases. Moreover, the regions of the enteric bacterial proteins which are contiguous with the short sequences shared with B27 tend to have structures which are also predicted to bind B27. These observations suggest a mechanism for autoimmunity and lead to the prediction that the B27-associated diseases are mediated by a subset of T-cell receptors, B27, and the peptides bound by B27. PMID- 8415703 TI - Rod photoreceptor cGMP-phosphodiesterase: analysis of alpha and beta subunits expressed in human kidney cells. AB - The bovine alpha and murine beta subunits of rod-photoreceptor cGMP phosphodiesterase (PDE alpha and PDE beta) were expressed in adenovirus transformed 293 human embryonic kidney cells. RNA blots from transfected cells showed transcripts of 3.0 and 2.8 kb corresponding to PDE alpha and PDE beta, respectively. Protein expression was analyzed by using affinity-purified antibodies against cGMP-PDE on immunoblots and by immunoprecipitation. PDE alpha and PDE beta exhibited the expected mobility (and thus apparent molecular size) and had cGMP hydrolytic activity. Reconstitution of the PDE alpha beta heterodimer with the expressed proteins increased by approximately 6-fold the activity of the individual alpha and beta subunits. Addition of expressed beta subunit to retinal extracts from 9- to 10-day-old rd/rd mice (which have only normal alpha and gamma subunits of rod cGMP-PDE and thus minimal activity) increased enzyme activity by approximately 3-fold. Our results therefore demonstrate that photoreceptor-specific cGMP-PDE can be synthesized in human kidney cells with consequent expression of enzymatic activity. PMID- 8415704 TI - The PML-retinoic acid receptor alpha translocation converts the receptor from an inhibitor to a retinoic acid-dependent activator of transcription factor AP-1. AB - We report here that the fusion of PML, a nuclear protein defined by the t(15;17) chromosomal translocation in acute promyelocytic leukemia, with retinoic acid receptor alpha (RAR alpha) changes the RAR alpha from a retinoic acid (RA) dependent inhibitor to a RA-dependent activator of AP-1 transcriptional activity. The PML-RAR alpha chimera cooperates with c-Jun and, strikingly, with c-Fos to stimulate the transcription of both synthetic and natural reporter genes containing an AP-1 site. Stimulation is dependent on the concentration of RA and its dose-response curve is comparable to that for activation by RAR alpha of transcription on RA-responsive genes. Further, in the absence of RA, a circumstance in which RAR alpha has no effect on AP-1 activity, PML-RAR alpha is an inhibitor. Deletion of the dimerization, transactivation, or DNA-binding domains of c-Jun and removal of the PML dimerization domain in the PML-RAR alpha hybrid abrogates their transcriptional cooperatively. In view of the association between AP-1 activity and hemopoietic differentiation, we suggest that these properties of PML-RAR alpha could contribute to the leukemic phenotype and its response to RA. PMID- 8415705 TI - Functional complementation of an Escherichia coli ribonuclease H mutation by a cloned genomic fragment from the trypanosomatid Crithidia fasciculata. AB - A gene designated Cfa RNH1 has been cloned by complementation of an RNase H deficiency in an Escherichia coli rnhA mutant by using a genomic DNA library from the trypanosomatid Crithidia fasciculata. The encoded RNase H is predicted to have 494 amino acid residues and a molecular mass of 53.7 kDa. The carboxyl half of the protein is homologous to the 155-residue E. coli RNase HI (41% identity) and the 166-residue Saccharomyces cerevisiae RNase HI (33% identity). The recombinant protein has been purified as a six-histidine-tagged fusion protein by metal chelate chromatography and was shown to have RNase H activity. Antibodies against the recombinant protein recognize proteins of approximately 65 kDa and 56 kDa on Western blots of C. fasciculata extracts. These results demonstrate the feasibility of cloning trypanosome genes by complementation of appropriate E. coli mutants with genomic DNA libraries. PMID- 8415706 TI - Tie-1 and tie-2 define another class of putative receptor tyrosine kinase genes expressed in early embryonic vascular system. AB - We report the molecular cloning and characterization of two structurally related putative receptor tyrosine kinases, encoded by distinct genes (tie-1 and tie-2) on mouse chromosome 4. Both tie-1 and tie-2 encode receptor proteins possessing unique multiple extracellular domains: two immunoglobulin-like loop domains flanking three epidermal growth factor repeats followed by three fibronectin-type III repeats. Both genes are expressed in early embryonic vascular system and in maternal decidual vascular endothelial cells, where the vasculature undergoes an active angiogenesis. tie-2, but not tie-1, expression was also detected in extraembryonic mesoderm of the amnion. tie-1, but not tie-2, is expressed in an acute myelogenic cell line in vitro. tie-1 and tie-2 may form another class within the receptor tyrosine kinase gene family, and further characterization of these genes and identification of their putative ligands should define the nature of the signal-transduction cascades underlying early vascular system development, as well as their differential roles in mesodermal cells of the amniotic and myeloid lineages. PMID- 8415707 TI - Cell-type-specific expression of rat steroid 5 alpha-reductase isozymes. AB - The enzyme steroid 5 alpha-reductase (EC 1.3.99.5) is a component of an intercellular signaling pathway that determines cell fate in the primordium of the mammalian reproductive tract. During male phenotypic sexual differentiation, the dihydrotestosterone product of this enzyme binds to the androgen receptor and initiates development of the external genitalia and prostate. Genes encoding two isozymes of steroid 5 alpha-reductase with different biochemical properties and tissue distributions have recently been isolated. In the current study, we utilize in situ hybridization analysis to determine cell-type-specific expression patterns of the 5 alpha-reductase isozyme mRNAs in two androgen target tissues (regenerating ventral prostate and epididymis) and a peripheral tissue (liver). In regenerating ventral prostate, the type 1 mRNA is expressed in basal epithelial cells whereas expression of the type 2 mRNA is largely confined to stromal cells. These results were confirmed by immunohistochemical analysis and are consistent with distinct roles played by the isozymes in the prostate. In the epididymis, both 5 alpha-reductase isozyme mRNAs are expressed in epithelial cells. Only the type 1 mRNA is present in the liver. This mRNA is distributed in a striking spatial gradient extending from hepatocytes surrounding the portal triad (high expression) to those surrounding the central vein (low to absent expression). These findings demonstrate cell-type-specific expression of the steroid 5 alpha-reductase isozymes and underscore their distinct and overlapping functions in androgen physiology. PMID- 8415708 TI - Twist constraints on linker DNA in the 30-nm chromatin fiber: implications for nucleosome phasing. AB - Previous work has shown that nucleosome repeat lengths, and hence linker DNA lengths, are preferentially quantized to a set of values differing by integral multiples of the helical twist of DNA. An explanation was proposed in which this preferential quantitation is due to twist constraints on linker DNA arising from nucleosome-nucleosome interactions in folded chromatin. Here we report the results of a study, using ethidium intercalation, designed to test whether twist constraints do indeed exist. Electron microscopy reveals that ethidium intercalation causes decondensation of dinucleosomes. Direct measurement of the free energy of intercalation by fluorescence spectroscopy reveals competition between chromatin folding and ethidium intercalation. Results from other laboratories establish that these effects of ethidium are due to ethidium-induced changes in the twist of linker DNA, and not to a variety of other possible effects. We conclude that twist constraints on linker DNA do exist. These may explain the observation of preferentially quantized linker DNA lengths. Implications of these results for mechanisms of nucleosome phasing and the mechanisms of drug action are discussed. PMID- 8415709 TI - JunD mutants with spontaneously acquired transforming potential have enhanced transactivating activity in combination with Fra-2. AB - Although a replication-competent retrovirus that carries junD has no transforming activity in chicken embryo fibroblasts, we have isolated mutant viruses that have spontaneously acquired transforming activity. The molecularly cloned junD genes of three such mutant viruses (T1, T2, and T3) were shown to be responsible for the cellular transformation. DNA sequence analysis indicated that a specific polynucleotide in the junD sequence was tandemly multiplied three times of five times in T1 and T2, respectively. The repeated polynucleotide encodes 16 amino acid residues that are located in a highly conserved region among Jun family proteins. The junD mutation in T3 involved an inversion, a translocation, and nucleotide substitutions that caused drastic amino acid exchanges in another well conserved region among Jun family proteins. The transcriptional activity of these mutants was analyzed by means of transient expression experiments in F9 cells using a reporter gene containing a single AP-1 binding site. Compared with the wild-type JunD, none of them showed enhanced transactivating activity in the forms of homodimers or of heterodimers with c-Fos or Fra-1. However, they did exhibit much higher transactivating activity than the wild type when they formed heterodimers with Fra-2, indicating that the mutated regions function as transactivation domains in a partner-specific manner. Since we have previously reported that there is a basal level of Fra-2 expression in chicken embryo fibroblasts, the results may indicate that protein complexes between JunD mutants and Fra-2 play a crucial role in the cellular transforming activity. PMID- 8415710 TI - Superfamily Gondwanatherioidea: a previously unrecognized radiation of multituberculate mammals in South America. AB - Multituberculates were the longest-lived order of the Class Mammalia and, during the Mesozoic and early Cenozoic, were among the most diverse and abundant representatives of the class. However, until the recent discovery of two Cretaceous teeth, one from South America and one from Africa, they were known only from northern continents. Additional material of the South American form Ferugliotherium has confirmed its multituberculate affinities and indicates that it may be a derived member of the Suborder Plagiaulacoidea. New specimens provide evidence that two other South American forms, Gondwanatherium and Sudamerica, are also multituberculates and that they are closely related to Ferugliotherium. Gondwanatherium and Sudamerica, each possessing highly specialized hypsodont molars, were previously thought to be the earliest known representatives of the Edentata, to be involved in the origin of edentates, or to represent a previously unknown higher taxon of mammals. However, there are detailed similarities in gross dental morphology, enamel microstructure, and inferred direction of jaw movement among Ferugliotherium, Gondwanatherium, and Sudamerica. All three genera are here regarded as representatives of a highly derived, endemic radiation of South American multituberculates and are allocated to the superfamily Gondwanatherioidea. Multituberculates were therefore more common elements of Late Cretaceous and early Paleocene mammalian faunas of South America than previously recognized. PMID- 8415711 TI - Transnuclear retrotransposition of the Tad element of Neurospora. AB - Tad is a LINE-like DNA element found in Neutrospora crassa. A Neurospora artificial intron based on the first intron of the am (glutamate dehydrogenase) gene was constructed and introduced, in the correct orientation, into a unique Nru I site in open reading frame 1 of an active Tad element, Tad1-1. Transformants containing the Tad element with the artificial intron were placed in forced heterokaryons with strains lacking Tad elements. Tad was shown to transpose between nuclei in these heterokaryons. Examination of the transposed Tad elements showed that the intron had been precisely removed in all cases. This confirms that Tad is a retrotransposon and that there is a cytoplasmic phase in these retrotransposition events. PMID- 8415712 TI - Identification of additional members of human G-protein-coupled receptor kinase multigene family. AB - Human neutrophils express several distinct guanine nucleotide binding (G)-protein coupled receptors that mediate their responsiveness to chemoattractants. Phosphorylation by receptor-specific and second messenger-activated protein kinases is a common mechanism for regulation of G-protein-coupled receptors. To explore the possibility that chemoattractant receptors are regulated by unique receptor kinases, we utilized PCR to identify receptor kinases in human neutrophils. Here, we report the isolation of three G-protein-coupled-receptor kinase (GPRK)-like sequences termed GPRK5, GPRK6, and GPRK7 in addition to the beta-adrenergic receptor kinase (beta ARK) 1 and 2 isoforms (beta ARK1 and beta ARK2). Two, GPRK5 and GPRK6, showed high homology at the amino acid level to the recently identified receptor-kinase-like sequence localized close to the Huntington disease locus. GPRK7 is of interest in that it contains a DLG (Asp-Leu Gly) amino acid motif of receptor kinases preceded by a DFD (Asp-Phe-Asp) motif. We isolated cDNAs corresponding to GPRK6; the complete sequence shows > 66% identity and 81% similarity at the amino acid level to the GPRK from the Huntington disease locus. The GPRK6 cDNA probe hybridizes to two mRNAs of 2.9 and 2.1 kb that were expressed in all the tested human tissues including HL-60 cells and neutrophils. Genomic Southern blot analysis and chromosome mapping showed that GPRK6 hybridizes to two closely related genes located on chromosomes 5 and 13 and are, therefore, distinct from the GPRK located near the Huntington disease locus on chromosome 4. The identification herein of three putative receptor kinases indicates that in addition to beta ARK and rhodopsin kinase subfamilies, there are other receptor-kinase subfamilies that regulate the broad spectrum of G protein-coupled receptors. PMID- 8415713 TI - Effects of Mn2+ and Mg2+ on assimilation of NO3- and NH4+ by soil microorganisms. AB - Although it has been demonstrated that Mn2+ and Mg2+ can influence the activity of glutamine synthetase in various organisms, there is little information concerning the effects of these cations on the activity of this enzyme in soil microorganisms or on ability of these microorganisms to assimilate NO3- and NH4+. We studied the effects of different concentrations of Mn2+ and Mg2+ on assimilatory NO3- reduction and NH4+ assimilation in cultures of two microorganisms commonly found in soil [Pseudomonas fluorescens (ATCC 13525) and Azotobacter chroococcum (ATCC 9043)] and in an enrichment culture of soil microorganisms. We found that Mn2+ strongly inhibited NH4+ assimilation by soil microorganisms and blocked the inhibitory effect of NH4+ on assimilatory NO3- reductase (ANR) activity, thereby uncoupling ANR activity from nitrogen assimilation and causing the NH4+ formed by ANR activity to be released to the environment. Mg2+ counteracted the effect of Mn2+ on microbial metabolism of nitrogen, which suggests that the overall effect of these cations on nitrogen assimilation by soil microorganisms will depend on the ratio of their concentrations in soil. PMID- 8415714 TI - Reproducing the three-dimensional structure of a tRNA molecule from structural constraints. AB - The three-dimensional structure of yeast tRNA(Phe) was reproduced at atomic resolution with the automated RNA modeling program MC-SYM, which is based on a constraint-satisfaction algorithm. Structural constraints used in the modeling were derived from the secondary structure, four tertiary base pairs, and other information available prior to the determination of the x-ray crystal structure of the tRNA. The program generated 26 solutions (models), all of which had the familiar "L" form of tRNA and root-mean-square deviations from the crystal structure in the range of 3.1-3.8 A. The interaction between uridine-8 and adenosine-14 was crucial in the modeling procedure, since only this among the tertiary pairs is necessary and sufficient to reproduce the L form of tRNA. Other tertiary interactions were critical in reducing the number of solutions proposed by the program. PMID- 8415715 TI - Early Cretaceous mammal from North America and the evolution of marsupial dental characters. AB - A mammal from the Early Cretaceous of the western United States, represented by a lower jaw exceptional in its completeness, presents unambiguous evidence of postcanine dental formula in an Early Cretaceous marsupial-like mammal, and prompts a reconsideration of the early evolution of marsupial dental characters. A marsupial postcanine dental formula (three premolars and four molars) and several marsupial-like features of the lower molars are present in the new taxon, but a hallmark specialization of marsupials (twinning of the hypoconulid and entoconid on lower molars) is lacking. This, coupled with recent evidence from the Late Cretaceous of the western United States, suggests that the distinctive marsupial dental formula evolved prior to the most characteristic specialization of lower molars and that apomorphies presumed to be diagnostic of the upper molars (such as auxiliary stylar cusps) were relatively more recent developments in marsupial history. Dental evidence supports the monophyly of higher (tribosphenic) mammals and suggests that the predominantly Old World Deltatheroida, recently proposed as a sister taxon to marsupials, represents a primitive and unrelated group of higher mammals; by this interpretation, early marsupials and their presumed close relatives are restricted to North America. This, together with the hypothesized relationships of South American/Australian marsupials (in the context of the North American Cretaceous radiation) and evidence from the fossil record of South America, in turn supports a North American origin for the group. PMID- 8415716 TI - Cloning and molecular characterization of a human intracellular serine proteinase inhibitor. AB - We describe a cDNA encoding a serine proteinase inhibitor present in placental tissue and the cytosolic fraction of K562 cells. On the basis of its interaction with thrombin, through which it was discovered, the inhibitor has been operationally named the placental thrombin inhibitor (PTI). Amino acid sequence comparisons suggest that its reactive center is located at Arg-341 and Cys-342, that it lacks a classical N-terminal signal sequence, and that it has the highest degree of similarity to intracellular serine proteinase inhibitors (serpins), such as the human monocyte/neutrophil elastase inhibitor and the equine leukocyte elastase inhibitor. PTI also resembles these inhibitors in that it contains oxidation-sensitive residues adjacent to the reactive site. The PTI cDNA was expressed in rabbit reticulocyte lysate and in COS-7 cells and a 42-kDa protein was produced. Recombinant PTI formed a 67-kDa complex when incubated with thrombin. The ability of native PTI to bind thrombin was destroyed by incubation with iodoacetamide. Analysis of human tissue mRNA indicated that PTI is expressed widely with the highest levels in cardiac and skeletal muscle and placenta. We conclude that PTI is a member of an emerging class of intracellular serpins. PMID- 8415717 TI - Significant enlargement of a specific subset of CD3+CD8+ peripheral blood leukocytes mediating cytotoxic T-lymphocyte activity during human immunodeficiency virus infection. AB - We have obtained a monoclonal antibody termed BY55 that defines an 80-kDa cell surface structure on a subset of circulating peripheral blood mononucleocytes. This structure, which was not detected on most cell lines or activated lymphocytes, was expressed exclusively on 15-25% of CD2+ circulating lymphocytes, including a major subset within the CD3- and the T-cell receptor gamma delta + lymphocytes and a small percentage of the CD3+CD8+ cells. Moreover, we have shown that the BY55 molecule delineated the competent killer circulating lymphocytes. In the present report, additional two- and three-color immunofluorescence studies of peripheral blood lymphocytes were done to precisely determine BY55 expression within the T-cell population. In normal individuals, peripheral blood CD3+CD8+BY55+ cells represented only 5-6% of the lymphocytes, and these cells possessed cytolytic activity. Interestingly, we found that the percentage of total BY55+ lymphocytes as well as the percentage of CD3+CD8+BY55+ was significantly increased in peripheral blood lymphocytes of human immunodeficiency virus-seropositive individuals. PMID- 8415718 TI - K+ channel openers prevent global ischemia-induced expression of c-fos, c-jun, heat shock protein, and amyloid beta-protein precursor genes and neuronal death in rat hippocampus. AB - Transient global forebrain ischemia induces in rat brain a large increase of expression of the immediate early genes c-fos and c-jun and of the mRNAs for the 70-kDa heat-shock protein and for the form of the amyloid beta-protein precursor including the Kunitz-type protease-inhibitor domain. At 24 hr after ischemia, this increased expression is particularly observed in regions that are vulnerable to the deleterious effects of ischemia, such as pyramidal cells of the CA1 field in the hippocampus. In an attempt to find conditions which prevent the deleterious effects of ischemia, representatives of three different classes of K+ channel openers, (-)-cromakalim, nicorandil, and pinacidil, were administered both before ischemia and during the reperfusion period. This treatment totally blocked the ischemia-induced expression of the different genes. In addition it markedly protected neuronal cells against degeneration. The mechanism of the neuroprotective effects involves the opening of ATP-sensitive K+ channels since glipizide, a specific blocker of that type of channel, abolished the beneficial effects of K+ channel openers. The various classes of K+ channel openers seem to deserve attention as potential drugs for cerebral ischemia. PMID- 8415719 TI - Molecular dissection of subunit interfaces in the acetylcholine receptor: identification of residues that determine curare selectivity. AB - The acetylcholine receptor from vertebrate skeletal muscle is a transmembrane channel that binds nerve-released acetylcholine to elicit rapid transport of small cations. Composed of two alpha subunits and one beta, one gamma, and one delta subunit, the receptor is a cooperative protein containing two sites that bind agonists, curariform antagonists, and snake alpha-toxins. Until recently the two binding sites were thought to reside entirely within each of the two alpha subunits, but affinity labeling and expression studies have demonstrated contributions by the gamma and delta subunits. Affinity labeling and mutagenesis studies have identified residues of the alpha subunit that contribute to the binding site, but the corresponding gamma- and delta-subunit residues remain unknown. By making gamma-delta chimeras and following the nearly 100-fold difference in curare affinity for the two binding sites, the present work identified residues of the gamma and delta subunits likely to be near the binding site. Two sets of binding determinants were identified in homologous positions of the gamma and delta subunits. The determinants lie on either side of a disulfide loop found within the major extracellular domain of the subunits. This loop is common to all acetylcholine, gamma-aminobutyrate, and glycine receptor subunits. PMID- 8415720 TI - Bacteriorhodopsin is involved in halobacterial photoreception. AB - The bacterio-opsin gene was introduced into a "blind" Halobacterium salinarium mutant that (i) lacked all the four retinal proteins [bacteriorhodopsin (BR), halorhodopsin, and sensory rhodopsins (SRs) I and II] and the transducer protein for SRI and (ii) showed neither attractant response to long wavelength light nor repellent response to short wavelength light. The resulting transformed cells acquired the capability to sense light stimuli. The cells accumulated in a light spot, demonstrating the BR-mediated orientation in spatial light gradients. As in wild-type cells, a decrease in the intensity of long wavelength light caused a repellent response by inducing reversals of swimming direction, but, in contrast to wild-type cells, a decrease in the intensity of short wavelength light also repelled the cells. An increase in light intensity evoked an attractant response (i.e., a transient suppression of spontaneous reversals). Signal processing times and adaptation kinetics were similar to the SRI-mediated reactions. However, compared to SR-mediated photoresponses, higher light intensities were necessary to induce the BR-mediated responses. The light sensitivity of the transformant was increased by adding 1 mM cyanide and decreased by the addition of arginine, agents that respectively reduce and increase the light-independent generation of the electrochemical potential difference of H+ ions (delta mu H+). A decrease in irradiance to an intensity that was still high enough to saturate BR-initiated delta mu H+ changes failed to induce the repellent effect, but the addition of a protonophorous uncoupler sensitized the cell to these light stimuli. The BR D96N mutant (Asp-96 is replaced by Asn) with decreased proton pump activity showed strongly reduced BR-mediated responses. Azide, which increases this mutant's H+ pump efficiency, increased the photosensitivity of the mutant cells. Moreover, azide diminished (i) the membrane potential decreasing and (ii) repellent effects of blue light added to the orange background illumination in this mutant. We conclude that the BR-mediated photoreception is due to the light-dependent generation of delta mu H+. Our data are consistent with the assumption that the H. salinarium cell monitors the membrane energization level with a "protometer" system measuring total delta mu H+ changes or its electric potential difference component. PMID- 8415721 TI - Electromigration behavior of polysaccharides in capillary electrophoresis under pulsed-field conditions. AB - Various polysaccharides can successfully migrate through entangled polymer solutions during high-voltage capillary electrophoresis. For neutral polysaccharides, complexation with borate provides the electric charge needed for electromigration, while a fluorescent tag is needed to detect the solute bands with adequate sensitivity. At constant potentials between 50 and 300 V/cm, the charged polysaccharides undergo molecular stretching, resisting the desired separation according to their molecular mass. This problem can be overcome through the use of variable fields, pulsed along the separation capillary at a 180 degree angle. Variables of the pulsing experiment appear to have a profound influence on molecular shape rearrangements of polysaccharides with respect to the separation medium, as demonstrated here with highly efficient separations of polydextrans (8,000-2,000,000 Da). PMID- 8415722 TI - The yeast Candida albicans has a clonal mode of reproduction in a population of infected human immunodeficiency virus-positive patients. AB - To ascertain the population structure of Candida albicans, we have carried out a multilocus enzyme electrophoresis study based on the analysis of 21 gene loci. We have thus characterized 55 strains isolated one each from 55 human immunodeficiency virus-positive patients. There is considerable polymorphism among the strains. A population-genetic analysis indicates that the two fundamental consequences of sexual reproduction (i.e., segregation and recombination) are apparently absent in this population of C. albicans. The population structure of C. albicans appears to be clonal, a state of affairs that has important medical and biological consequences. PMID- 8415723 TI - Estrogen receptor and progesterone receptor genes are expressed differentially in mouse embryos during preimplantation development. AB - Estrogen and progesterone play an important role in the development and implantation of preimplantation embryos. However, it is controversial whether these hormones act directly on the embryos. The effects of these hormones depend on the existence of their specific receptors. To determine whether estrogen receptor (ER) and progesterone receptor genes are expressed in mouse preimplantation embryos, we examined RNA from embryos at different stages of preimplantation development by reverse transcription-polymerase chain reaction techniques. ER mRNA was found in oocytes and fertilized eggs. The message level began to decline at the two-cell stage and reached its lowest level at the five- to eight-cell stage. ER mRNA was not detectable at the morula stage but reappeared at the blastocyst stage. Progesterone receptor mRNA was not detectable until the blastocyst stage. The embryonic expression of ER and progesterone receptor genes in the blastocyst suggests a possible functional requirement for ER and progesterone receptor at this stage of development. These results provide a basis for determining the direct role of estrogen and progesterone in preimplantation embryos. PMID- 8415724 TI - High-mobility-group 1 protein mediates DNA bending as determined by ring closures. AB - High-mobility-group 1 protein (HMG1) is an abundant eukaryotic DNA-binding protein, the cellular role of which remains ill-defined. To test the ability of HMG1 itself to mediate curvature in double-stranded DNA, we examined its effect on the phage T4 DNA ligase-dependent cyclization of short DNA fragments. HMG1 caused circle formation for fragments > or = 87 bp. Fragments of 123, 100, 92, and 87 bp did not cyclize in the absence of protein but formed covalently closed circular monomers efficiently in the presence of HMG1, indicating that the protein is capable of introducing bends into the duplex. The bending activity was maintained by a 79-amino acid polypeptide corresponding to a single HMG-box domain of HMG1. The binding affinity for the DNA minicircle was greater than for the corresponding linear fragment. These findings indicate that the role of HMG1 could involve both structure-specific recognition of prebent DNA and distortion of the DNA helix by bending and that the HMG-box domain may actually be responsible for this activity. PMID- 8415725 TI - Identification in the HLA class I region of a gene expressed late in keratinocyte differentiation. AB - A gene designated S has been identified in the class I region of the human major histocompatibility complex. The S gene is located 160 kb telomeric of HLA-C. It is expressed at high levels as 2.2-kb and 2.6-kb mRNAs in human skin. No homologous transcripts were detected in other tissues including placenta, liver, spleen, thymus, and brain. In situ hybridization showed that S gene expression was restricted to the differentiating keratinocytes in the granular layer of the epidermis. The predicted amino acid sequence of the S protein was remarkable for its high content of serine, glycine, and proline. There were significant similarities with the amino acid sequences of loricrin, keratin 1, and keratin 10, all major components of the granular-cell layer. The selective expression of the S gene in the granular-cell layer in the epidermis suggests a role in the developmental program of differentiating keratinocytes. Furthermore, in light of the recognized association of psoriasis vulgaris, a disorder of keratinocyte proliferation, with alleles of HLA-C, this gene may contribute primarily to the pathogenesis of this common disorder. PMID- 8415726 TI - Identification of a Drosophila activin receptor. AB - Activins are cytokines of the transforming growth factor beta superfamily that control various events during vertebrate embryo development and cell differentiation in the adult, and act through transmembrane receptors that contain a cytoplasmic protein-serine/threonine kinase domain. We describe the identification, deduced primary structure, and expression pattern of Atr-II, a receptor serine/threonine kinase found in Drosophila. With the exception of the spacing of 10 cysteine residues, the extracellular domain of Atr-II is very dissimilar from those of vertebrate activin receptors, yet it binds activin with high affinity and specificity. The kinase domain sequence of Atr-II is 60% identical to those of activin receptors from vertebrates, suggesting similarities in their signaling mechanisms. Maternal Atr-II transcript and its product are abundant in the oocyte. During development, the highest levels of Atr-II transcript and protein are observed in the mesoderm and gut. The possible role of an activin signaling system in Drosophila development is discussed. PMID- 8415727 TI - Measurement of the g-tensor of the P700+. signal from deuterated cyanobacterial photosystem I particles. AB - We report high-field continuous wave EPR spectra of P700+. in preparations obtained from deuterated cyanobacteria (Synechococcus lividus). Measurements were performed with photosystem I (PS-I) preparations, whole cells from cyanobacteria grown in 2H2O, and photosystem II (PS-II) preparations, as well as with protonated PS-I preparations. Because of the significantly improved resolution of our 140-GHz spectrometer (as compared with X- or Q-band EPR) the principal values of the g-tensor of the primary donor P700+. could be resolved and measured with high accuracy as g11 = 2.00304, g22 = 2.00262, and g33 = 2.00232. Other signals arising from Mn2+ and a dark signal from PS-II at g approximately 2.00266 are distinguished from the P700+. g-tensor powder pattern. The measured g values are compared with those of several bacterial reaction center donors. PMID- 8415728 TI - Chromosomal rearrangement segregating with adrenoleukodystrophy: a molecular analysis. AB - The relationship between X chromosome-linked adrenoleukodystrophy and the red/green color pigment gene cluster on Xq28 was investigated in a large kindred. The DNA in a hemizygous male showed altered restriction fragment sizes compatible with at least a deletion extending from the 5' end of the color pigment genes. Segregation analysis using a DNA probe within the color pigment gene cluster showed significant linkage with adrenoleukodystrophy (logarithm of odds score of 3.19 at theta = 0.0). These data demonstrate linkage, rather than association, between a unique molecular rearrangement in the color pigment gene cluster and adrenoleukodystrophy. The DNA changes in this region are thus likely to be helpful for determining the location and identity of the responsible gene. PMID- 8415729 TI - Chromosomal rearrangement segregating with adrenoleukodystrophy: associated changes in color vision. AB - A patient from a large kindred with adrenoleukodystrophy showed profound disturbance of color ordering, color matching, increment thresholds, and luminosity. Except for color matching, his performance was similar to blue-cone "monochromacy," an X chromosome-linked recessive retinal dystrophy in which color vision is dichromatic, mediated by the visual pigments of rods and short-wave sensitive cones. Color matching, however, indicated that an abnormal rudimentary visual pigment was also present. This may reflect the presence of a recombinant visual pigment protein or altered regulation of residual pigment genes, due to DNA changes--deletion of the long-wave pigment gene and reorganized sequences 5' to the pigment gene cluster--that segregate with the metabolic defect in this kindred. PMID- 8415730 TI - Expression of alpha-thymosins in human tissues in normal and abnormal growth. AB - Radioimmunoassays specific for the N and C termini of human prothymosin alpha and the N terminus of human parathymosin alpha were employed for the measurement of the levels of alpha-thymosins in human thymus, spleen, and liver during normal growth and intestine and breast in malignant growth. A differential expression of the two alpha-thymosins was observed in thymus (prothymosin alpha-rich) and liver (parathymosin alpha-rich). A decline in the levels of both alpha-thymosins was found with age, with prothymosin alpha in thymus showing the sharpest change (15- to 30-fold). The levels of both alpha-thymosins were higher in malignant tissues as compared with healthy ones. In breast cancer, in particular, the mean increase for prothymosin alpha and parathymosin alpha was 17.9- and 11.5-fold, respectively. The major crossreactive material was characterized in all cases as intact prothymosin alpha and parathymosin alpha. These results suggest an in vivo relationship of the expression of alpha-thymosins with the human tissue cell proliferation activity. PMID- 8415731 TI - Molecular characterization of monoclonal CRIA-positive anti-arsonate antibodies derived from idiotype-negative mice bearing a light chain polymorphism. AB - We have elicited anti-arsonate antibodies bearing the major cross-reactive idiotype (CRIA) in a double congenic idiotype-negative strain (C.C58.AL-20) bearing a light chain polymorphism that has previously been shown serologically not to complement idiotype-positive heavy chains. Using the idiotype cascade (Ab1 ->Ab2-->Ab3-->-->Ab1'), CRIA-positive antibodies were raised and monoclonal antibodies were isolated and characterized serologically and by nucleotide sequence analysis. Two types of idiotype-positive anti-arsonate antibodies were generated in the C.C58.AL-20 strain. One group of hybridomas used the canonical VH1.8 heavy chain gene segment with V kappa 10 variant light chains. A second group used a VHGAM3.8 heavy chain with V kappa 10 variant light chains. This latter heavy-light pairing has been observed in CRIA-like responses previously in BALB/c mice after idiotypic manipulation (or rarely after antigen alone). These studies demonstrate the plasticity of the immune response when manipulated with idiotype reagents as well as its structural variability. Additionally, they provide important insights into the potentials of idiotype vaccines. PMID- 8415732 TI - Human neurons derived from a teratocarcinoma cell line express solely the 695 amino acid amyloid precursor protein and produce intracellular beta-amyloid or A4 peptides. AB - The beta-amyloid or beta/A4 peptides that accumulate as filamentous aggregates in the extracellular space of Alzheimer disease (AD) brains are derived from one or more alternatively spliced amyloid precursor proteins (APPs). The more abundant APPs in the central nervous system are the 695-(APP695), 751- (APP751), and 770- (APP770) amino acid isoforms, and each could be the source of beta/A4 peptide that accumulates in the AD brain. It is plausible that altered metabolism of these APPs by central nervous system neurons could lead to the release and deposition of beta/A4 peptide in brain parenchyma. Thus, we examined the expression and processing of the three major brain APPs in nearly pure human neurons (NT2N cells) derived from a teratocarcinoma cell line (NTera2/c1.D1 or NT2 cells) after retinoic acid treatment. NT2N neurons expressed almost exclusively APP695, whereas NT2 cells expressed predominantly APP751/770. Furthermore, the processing of the APPs in NT2N cells was distinct from NT2 and nonneuronal cells. Most significantly, the NT2N neurons but not the NT2 cells constitutively generated intracellular beta/A4 peptide and released it into the culture medium. This work demonstrates the intracellular production of beta/A4 peptide and suggests that cultured NT2N cells may provide a unique model system for understanding the contribution of neurons and APP695 to amyloidogenesis in the AD brain. PMID- 8415733 TI - Multiple Ca2+ channel types coexist to regulate synaptosomal neurotransmitter release. AB - The regulation of excitation-secretion coupling by Ca2+ channels is a fundamental property of the nerve terminal. Peptide toxins that block specific Ca2+ channel types have been used to identify which channels participate in neurotransmitter release. Subsecond measurements of [3H]-glutamate and [3H]dopamine release from rat striatal synaptosomes showed that P-type channels, which are sensitive to the Agelenopsis aperta venom peptide omega-Aga-IVA, trigger the release of both transmitters. Dopamine (but not glutamate) release was also controlled by N-type, omega-conotoxin-sensitive channels. With strong depolarizations, where neither toxin was very effective alone, a combination of omega-Aga-IVA and omega conotoxin produced a synergistic inhibition of 60-80% of Ca(2+)-dependent dopamine release. The results suggest that multiple Ca2+ channel types coexist to regulate neurosecretion under normal physiological conditions in the majority of nerve terminals. P- and N-type channels coexist in dopaminergic terminals, while P-type and a omega-conotoxin- and omega-Aga-IVA-resistant channel coexist in glutamatergic terminals. Such an arrangement could lend a high degree of flexibility in the regulation of transmitter release under diverse conditions of stimulation and modulation. PMID- 8415734 TI - A light stabilizer (Tinuvin 770) that elutes from polypropylene plastic tubes is a potent L-type Ca(2+)-channel blocker. AB - A pharmacologically active agent was easily extracted by aqueous or organic solvents from laboratory plastic tubes (Falcon Blue Max) and has been chemically identified as bis(2,2,6,6-tetramethyl-4-piperidyl) sebacate. This compound (approximately 12 micrograms per tube approximately 25 nmol) blocked 1,4 dihydropyridine-sensitive 45Ca2+ uptake into GH3 cells with an IC50 value of 3.6 microM, inhibited Sr2+ currents through L-type Ca2+ channels in A7r5 smooth muscle cells in whole-cell patch-clamp experiments after extracellular application, and affected the high-affinity binding of Ca2+ entry-blocker ligands to a variety of preparations. Bis(2,2,6,6-tetramethyl-4-piperidyl) sebacate is a highly potent (IC50 values < 10 nM) inhibitor at the phenylalkylamine- and benzothiazepine-selective drug-binding domains of the alpha 1 subunit of L-type Ca2+ channels. This compound behaves as a heterotropic allosteric regulator for the 1,4-dihydropyridine-selective domain in purified Ca(2+)-channel preparations from rabbit skeletal muscle. (+)-Tetrandrine stimulation of 1,4-dihydropyridine binding to the membrane-bound L-type Ca2+ channel is inhibited by the compound in a competitive manner (Ki value = 6.8 nM). Bis(2,2,6,6-tetramethyl-4-piperidyl) sebacate is therefore classified as the prototype of another class of L-type Ca(2+)-channel blockers that binds to the alpha 1 subunit at the drug-binding domains selective for (+)-tetrandrine or (+)-cis-diltiazem. This compound is identical to Tinuvin 770, which is used worldwide as a light stabilizer for polyolefins. PMID- 8415735 TI - Herpes simplex virus infected cell polypeptide 4 preferentially represses Sp1 activated over basal transcription from its own promoter. AB - Herpes simplex virus type 1 infected cell polypeptide 4 (HSV-1 ICP4) is a multifunctional phosphoprotein that is essential for viral infection. It is both a repressor and an activator of viral gene expression depending upon the promoter. ICP4 represses transcription from its own promoter. In the present study, we used general transcription factors from HeLa cell nuclear extracts, recombinant TATA binding protein (TBP) and TFIIB, and the transcriptional activator Sp1 to reconstitute in vitro transcription for the ICP4 promoter and to examine the effects of purified ICP4 on transcription. ICP4 was able to effectively repress Sp1-induced transcription from ICP4 promoter templates that contain one or multiple Sp1 binding sites. The observed inhibition required the ICP4 binding site that spans the transcription initiation site. ICP4 did not inhibit basal transcription as inferred by its inability to inhibit transcription when (i) Sp1 was not included in transcription reactions, (ii) the templates contained no Sp1 binding sites, and (iii) TBP was used in place of TFIID in the reactions. The in vitro observations were consistent with the behavior of the same constructs expressed in cells from the herpes simplex virus type 1 genome. DNase I footprinting experiments revealed that ICP4 could co-occupy the ICP4 promoter region with TBP-TFIIB, indicating that ICP4 does not necessarily exclude these factors from binding to the TATA region. The data suggest that the repressive effects of ICP4 observed in this study result from ICP4 interfering with the interactions contributing to Sp1-induced transcription. PMID- 8415736 TI - Purification and characterization of a soluble salicylic acid-binding protein from tobacco. AB - Previously, we identified a soluble salicylic acid (SA)-binding protein (SABP) in tobacco whose properties suggest that it may play a role in transmitting the SA signal during plant defense responses. This SA-binding activity has been purified 250-fold by conventional chromatography and was found to copurify with a 280-kDa protein. Monoclonal antibodies capable of immunoprecipitating the SA-binding activity also immunoprecipitated the 280-kDa protein, indicating that it was responsible for binding SA. These antibodies also recognized the 280-kDa protein in immunoblots of the partially purified SABP fraction or the crude extract. However, when the crude extract was prepared in the presence of antioxidants, only a 57-kDa protein was recognized. Since the SABP has a native molecular mass of 240 kDa, it appears that the SABP is a complex which contains a 57-kDa subunit and perhaps one or more additional proteins which are covalently crosslinked in the absence of antioxidants. The ability of a variety of phenolic compounds to compete with SA for binding to the SABP was both qualitatively and quantitatively correlated with their biological activity in inducing defense-related genes. Moreover, the inducibility of the pathogenesis-related (PR)-1 genes by SA was proportional to the abundance of the SABP in different organs. These correlations are consistent with a role for the SABP in perceiving and transducing the SA signal in plant defense. PMID- 8415737 TI - An accuracy center in the ribosome conserved over 2 billion years. AB - The accuracy of translation in Escherichia coli is profoundly influenced by three interacting ribosomal proteins, S12, S4, and S5. Mutations at lysine-42 of S12, originally isolated as causing resistance to streptomycin, increase accuracy. Countervailing "ribosomal ambiguity mutations" (ram) in S4 or S5 decrease accuracy. In the eukaryotic ribosome of Saccharomyces cerevisiae, mutations in SUP46 and SUP44, encoding the proteins equivalent to S4 and S5, lead to omnipotent suppression--i.e., to less accurate translation. The evolution of ribosomal protein S12 can be traced, by comparison with archaebacteria and Tetrahymena, to S28 of S. cerevisiae, even though the two proteins share only very limited regions of homology. However, one region that has been conserved contains a lysine residue whose mutation leads to increased accuracy in E. coli. We have introduced into S28 of yeast the same amino acid substitutions that led to the original streptomycin-resistant mutations in E. coli. We find that they have a profound effect on the accuracy of translation and interact with SUP44 and SUP46, just as predicted from the E. coli model. Thus, the interplay of these three proteins to provide the optimal level of accuracy of translation has been conserved during the 2 billion years of evolution that separate E. coli from S. cerevisiae. PMID- 8415738 TI - Peptide nucleic acid (PNA) conformation and polymorphism in PNA-DNA and PNA-RNA hybrids. AB - Two hydrogen-bonding motifs have been proposed to account for the extraordinary stability of polyamide "peptide" nucleic acid (PNA) hybrids with nucleic acids. These interresidue- and intraresidue-hydrogen-bond motifs were investigated by molecular mechanics calculations. Energy-minimized structures of Watson-Crick base-paired decameric duplexes of PNA with A-, B-, and Z-DNA and A-RNA polymorphs indicate that the inherent stability of the complementary PNA helical structures is derived from interresidue, rather than from intraresidue, hydrogen bonds in all hybrids studied. Intraresidue-hydrogen-bond lengths are consistently longer than interresidue hydrogen bonds. Helical strand stability with interresidue hydrogen bond stabilization follows the order: B-(DNA.PNA) > A-(DNA.PNA) congruent to A-RNA.PNA > Z-(DNA.PNA). In the triplex hybrids A-(RNA.PNA2) and B (DNA.PNA2), differences between stabilities of the two decamers of thyminyl PNA with lysine amide attached to the C terminus (pnaT)10 strands are small. The Hoogsteen (pnaT)10 strands are of slightly higher potential energy than are the Watson-Crick (pnaT)10 strands. Antiparallel arrangement of PNAs in the triplex is slightly favored over the parallel arrangement based on the calculations. Examination by molecular mechanics of the PNA.DNA analogue of the NMR-derived structure for the B-double-stranded DNA dodecamer d(CG-CAAATTTGCG)2 in solution suggests that use of all bases of the genetic alphabet should be possible without loss of the specific interresidue-hydrogen-bonding pattern within the PNA strand. PMID- 8415739 TI - Simulation of the kinetics of ligand binding to a protein by molecular dynamics: geminate rebinding of nitric oxide to myoglobin. AB - We have begun to use molecular dynamics to simulate the kinetics of nitric oxide rebinding to myoglobin after photodissociation. Rebinding was simulated using a potential function that switches smoothly between a nonbinding potential and a binding potential as a function of the position and orientation of the ligand, with no barrier arising from the crossing of potential surfaces of different electron spin. In 96 of 100 trajectories, the ligand rebound in < 15 ps. The kinetic progress curve was obtained by determining the time in each trajectory at which the ligand rebound and then calculating the fraction of unbound ligands as a function of time. The curve can be well reproduced by a simple model based on the dynamics of a Langevin particle moving on a one-dimensional potential of mean force calculated from nonreactive protein trajectories. The rate of escape from the energy well adjacent to the heme is in good agreement with the value calculated from experimental data, suggesting that a multiple-well model provides a plausible explanation for the nonexponential rebinding kinetics. A transition state analysis suggests that protein conformational relaxation coupled to the displacement of the iron from the heme plane is an unlikely cause for the nonexponential rebinding of nitric oxide. PMID- 8415740 TI - Mitochondrial DNA sequence evolution in the Arctoidea. AB - Some taxa in the superfamily Arctoidea, such as the giant panda and the lesser panda, have presented puzzles to taxonomists. In the present study, approximately 397 bases of the cytochrome b gene, 364 bases of the 12S rRNA gene, and 74 bases of the tRNA(Thr) and tRNA(Pro) genes from the giant panda, lesser panda, kinkajou, raccoon, coatimundi, and all species of the Ursidae were sequenced. The high transition/transversion ratios in cytochrome b and RNA genes prior to saturation suggest that the presumed transition bias may represent a trend for some mammalian lineages rather than strictly a primate phenomenon. Transversions in the 12S rRNA gene accumulate in arctoids at about half the rate reported for artiodactyls. Different arctoid lineages evolve at different rates: the kinkajou, a procyonid, evolves the fastest, 1.7-1.9 times faster than the slowest lineage that comprises the spectacled and polar bears. Generation-time effect can only partially explain the different rates of nucleotide substitution in arctoids. Our results based on parsimony analysis show that the giant panda is more closely related to bears than to the lesser panda; the lesser panda is neither closely related to bears nor to the New World procyonids. The kinkajou, raccoon, and coatimundi diverged from each other very early, even though they group together. The polar bear is closely related to the spectacled bear, and they began to diverge from a common mitochondrial ancestor approximately 2 million years ago. Relationships of the remaining five bear species are derived. PMID- 8415741 TI - A single-base-pair mutation changes the specificities of both a transcription activation protein and its binding site. AB - The C1 protein of bacteriophage P22 binds to a unique site in the -35 region of the PRE promoter and activates transcription of the phage c2 repressor gene. This -35 target has an approximate direct repeat that overlaps the 5' end of the c1 coding region. We have isolated a single-base-pair mutation in this region that changes the PRE -35 target as well as the amino-terminal region of the C1 protein. Although the mutant C1 protein activates the mutant PRE promoter, it fails to activate the wild-type PRE promoter. This suggests that a single-base pair mutation changes the specificities of both a protein and its target site. These studies also indicate that C1 binding to DNA is influenced by contacts made through residues near the amino terminus. PMID- 8415742 TI - An Abd transgene prevents diabetes in nonobese diabetic mice by inducing regulatory T cells. AB - Susceptibility to the human autoimmune disease insulin-dependent diabetes mellitus is strongly associated with particular haplotypes of the major histocompatibility complex (MHC). Similarly, in a spontaneous animal model of this disease, the nonobese diabetic (NOD) mouse, the genes of the MHC play an important role in the development of diabetes. We have produced transgenic NOD mice that express the class II MHC molecule I-Ad in addition to the endogenous I Ag7 molecules in order to study the role of these molecules in the disease process. Although the inflammatory lesions within the islets of Langerhans in the pancreas appear similar in transgenic and nontransgenic animals, transgenic mice develop diabetes with greatly diminished frequency compared to their nontransgenic littermates (10% of transgenic females by 30 weeks of age compared to 45% of nontransgenic females). Furthermore, adoptive transfer experiments show that T cells present in the transgenic mice are able to interfere with the diabetogenic process caused by T cells from nontransgenic mice. Thus, the mechanism by which I-Ad molecules protect mice from diabetes includes selecting in the thymus and/or inducing in the periphery T cells capable of inhibiting diabetes development. PMID- 8415743 TI - Regulation of G1/S transition by cyclins D2 and D3 in hematopoietic cells. AB - Identification of the genes that control passage through the G1 phase of the cell cycle in mammalian cells is of particular interest because virtually all external events that regulate proliferation act primarily or exclusively during G1. Cyclins are likely to play a key role in controlling cell cycle progression, although their role during G1 in higher eukaryotic cells is unclear. In the hematopoietic cell line 32Dcl3, both cyclins D2 and D3 were expressed in proliferating cells, while cyclin D1 was undetectable. Expression of D2, and to a lesser extent D3, was interleukin 3 (IL-3) dependent and declined rapidly in the absence of this growth factor. To investigate the potential role of D cyclins in regulating cell growth, cell lines overexpressing either D2 or D3 were generated by transfection. Constitutive overexpression of either D2 or D3 did not affect cell viability, rate of cell proliferation, or dependence on IL-3 for growth. However, the distribution of cells through the cell cycle was dramatically altered, with both cyclins causing an increase in the fraction of cells in S phase, apparently related to a shortening of G1. Also, when deprived of IL-3, D3 overexpressing cells failed to arrest in G1, and apoptotic cell death in the absence of IL-3 was delayed. These results suggest a role for cyclins D2 and D3 in controlling passage of hematopoietic cells through G1 in the presence of growth factors and in effecting G1 arrest in the absence of growth factors. PMID- 8415744 TI - The conformation of the sarcin/ricin loop from 28S ribosomal RNA. AB - The sarcin/ricin loop is a highly conserved sequence found in the RNA of all large ribosomal subunits. The cytotoxins alpha-sarcin and ricin both inactivate ribosomes by cleaving a single bond in that loop. Once it has been attacked, ribosomes no longer interact with elongation factors properly, and translation stops. We have determined the conformation of the sarcin/ricin loop by multinuclear NMR spectroscopy using E73, a 29-nucleotide RNA that has the sarcin/ricin loop sequence and that is sensitive to both toxins in vitro. The sarcin/ricin loop has a compact structure that contains several purine.purine base pairs, a GAGA tetraloop, and a bulged guanosine adjacent to a reverse Hoogsteen A.U pair. It is stabilized by an unusual set of cross-strand base stacking interactions and imino proton to phosphate oxygen hydrogen bonds. In addition to having interesting structural features, this model explains many of the biochemical observations made about the loop's structure and its reactivity with cytotoxins, and it sheds light on the loop's interactions with elongation factors. PMID- 8415745 TI - Tissue-specific light-regulated expression directed by the promoter of a C4 gene, maize pyruvate,orthophosphate dikinase, in a C3 plant, rice. AB - Pyruvate,orthophosphate dikinase (PPDK; EC 2.7.9.1) activity is abundant in leaves of C4 plants, while it is difficult to detect in leaves of C3 plants. Recent studies have indicated that C3 plants have a gene encoding PPDK, with a structure similar to that of PPDK in C4 plants. However, low expression makes PPDK detection difficult in C3 plants. This finding suggests that high PPDK expression in C4 plants is due to regulatory mechanisms which are not operative in C3 plants. We have introduced a chimeric gene consisting of the gene encoding beta-glucuronidase (GUS; EC 3.2.1.31) controlled by the PPDK promoter from a C4 plant, maize, into a C3 cereal, rice. The chimeric gene was exclusively expressed in photosynthetic organs, leaf blades and sheaths, and not in roots or stems. Histochemical analysis of GUS activity demonstrated high expression of the chimeric gene in photosynthetic organs, localized in mesophyll cells, and no or very low activity in other cells. GUS expression was also regulated by light in that it was low in etiolated leaves and was enhanced by illumination. These observations indicate that the mechanisms responsible for cell-specific and light inducible regulation of PPDK observed in C4 plants are also present in C3 plants. We directly tested whether rice has DNA-binding protein(s) which interact with a previously identified cis-acting element of the C4-type gene. Gel retardation assays indicate the presence in rice of a protein which binds this element and is similar to a maize nuclear protein which binds PPDK in maize. Taken together, these results indicate that the regulatory system which controls PPDK expression in maize is not unique to C4 plants. PMID- 8415746 TI - Two tropinone reductases with different stereospecificities are short-chain dehydrogenases evolved from a common ancestor. AB - In the biosynthetic pathway of tropane alkaloids, tropinone reductase (EC 1.1.1.236) (TR)-I and TR-II, respectively, reduce a common substrate, tropinone, stereospecifically to the stereoisomeric alkamines tropine and pseudotropine (psi tropine). cDNA clones coding for TR-I and TR-II, as well as a structurally related cDNA clone with an unknown function, were isolated from the solanaceous plant Datura stramonium. The cDNA clones for TR-I and TR-II encode polypeptides containing 273 and 260 amino acids, respectively, and when these clones were expressed in Escherichia coli, the recombinant TRs showed the same strict stereospecificity as that observed for the native TRs that had been isolated from plants. The deduced amino acid sequences of the two clones showed an overall identity of 64% in 260-amino acid residues and also shared significant similarities with enzymes in the short-chain, nonmetal dehydrogenase family. Genomic DNA-blot analysis detected the TR-encoding genes in three tropane alkaloid-producing solanaceous species but did not detect them in tobacco. We discuss how the two TRs may have evolved to catalyze the opposite stereospecific reductions. PMID- 8415747 TI - Single amino acid substitutions uncouple the DNA binding and strand scission activities of Fok I endonuclease. AB - Single alanine substitution mutations at Asp-450 or Asp-467 of the type IIS restriction enzyme Fok I have no effect on the ability of the enzyme to bind strongly and selectively to its recognition site but completely eliminate its ability to cleave either strand of substrate DNA. Since wild-type Fok I shows no kinetic preference or required order of strand cleavage, these results indicate that Fok I, which evidently functions as a monomer, uses a single catalytic center to cleave both strands of DNA. In this respect, Fok I may resemble other monomeric enzymes that cleave double-stranded DNA. PMID- 8415748 TI - A 30-kDa alternative translation product of the CCAAT/enhancer binding protein alpha message: transcriptional activator lacking antimitotic activity. AB - Full-length (42 kDa) CCAAT/enhancer binding protein alpha (C/EBP alpha) (p42) has been implicated in the transcriptional activation of adipocyte genes including the 422(aP2) and C/EBP alpha genes during differentiation of 3T3-L1 preadipocytes. We have identified a 30-kDa isoform (p30) of C/EBP alpha that is expressed by 3T3-L1 adipocytes, mouse adipose tissue, and rat liver. In vitro translation of wild-type C/EBP alpha mRNA or transient transfection with a wild type C/EBP alpha vector gave rise to similar levels of p42 and p30. Mutational analysis revealed that p30 is an alternative translation product initiated at the third in-frame methionine codon of the C/EBP alpha message. p30C/EBP alpha binds to the C/EBP sites within and activates reporter gene expression driven by the 422(aP2) and C/EBP alpha gene promoters. Although transfection of 3T3-L1 preadipocytes with a strong p30C/EBP alpha expression vector is insufficient to induce differentiation, this vector advances the differentiation program. Unlike p42C/EBP alpha, which inhibits cell proliferation, p30C/EBP alpha is not antimitotic. Thus, the N-terminal 12-kDa segment of full-length C/EBP alpha contains an amino acid sequence necessary for antimitotic activity. During differentiation of 3T3-L1 preadipocytes and during hepatocyte development, the cellular p42C/EBP alpha/p30C/EBP alpha ratio changes, raising the possibility of a regulatory role. PMID- 8415749 TI - Tumor necrosis factor-induced c-myc expression in the absence of mitogenesis is associated with inhibition of adipocyte differentiation. AB - Tumor necrosis factor (TNF) inhibits and reverses differentiation of mouse adipogenic TA1 cells. We have found that TNF induces c-myc in a sustained manner in both preadipocytes and adipocytes; in contrast, serum induces c-myc transiently and only in preadipocytes. This TNF-mediated c-myc induction is not coupled with cell proliferation but is correlated with TNF-mediated inhibition of adipocyte differentiation. We prepared an inducible c-myc transformant of TA1 cells by transfection of the mouse c-myc gene under the control of the metallothionein-I promoter. These cells are unable to differentiate to adipocytes in the presence of Zn2+/Cd2+, and in differentiated TA1 cells, Zn2+/Cd2+ causes reduction of adipocyte-specific gene expression as does TNF. Lastly, exposure of TA1 cells to antisense c-myc oligonucleotide partially blocked the TNF-mediated reduction of adipocyte-specific gene expression. Thus, TNF-mediated c-myc expression is distinct in character from that involved in mitogenic responses but appears to play an important role in inhibition and reversal of adipocyte differentiation. PMID- 8415750 TI - PCR analysis of DNA from 70-year-old sections of rodless retina demonstrates identity with the mouse rd defect. AB - Rodless retina (gene symbol, r) was discovered in mice by Keeler 70 years ago and was first described in this journal as an autosomal recessive mutation leading to "the absence of the visual cells (rods), the external nuclear layer, and the external molecular layer" [Keeler, C. E. (1924) Proc. Natl. Acad. Sci. USA 10, 329-333]. The mutation was studied by Keeler and others in the United States and Europe over the next decade, but Keeler's stock was destroyed in 1939, and mice definitively related to his by pedigree and progeny tests also appeared to have been lost by the end of World War II. In the early 1950s Bruckner in Basel recognized mice with a similar retinal phenotype. Investigators in London and Strasbourg analyzed descendants of Bruckner's mice and concluded, on the basis of different pathogenesis from r, that they carried a new mutation, which came later to be called retinal degeneration, rd. The relationship of r and rd has been unsettled ever since. Now that the rd phenotype is known to be due to a nonsense mutation in the rod photoreceptor cGMP phosphodiesterase beta-subunit gene, we hoped to settle the question by direct analysis of r DNA. DNA was liberated from 70-year-old histological sections of +/r and r/r eyes, the only extant r DNA, and the regions encompassing the nonsense mutation amplified by the polymerase chain reaction (PCR). Sequence analysis of the PCR products revealed the presence of the same nonsense mutation and two intron polymorphisms in r DNA. PCR and direct sequence analysis of 11 strains of mice known to carry rd (or a similar allele) also revealed the presence of the nonsense mutation and the same intron polymorphisms. The fact that all r and rd mice contain an identical defect and intron polymorphisms in the phosphodiesterase beta-subunit gene settles beyond reasonable doubt that a single mutation arising > 70 years ago is now widely distributed through inbred mouse strains. Because of the extensive use of the name in publications of the past 40 years, we propose that the gene continue to be designated retinal degeneration, rd. PMID- 8415751 TI - Hematopoietic stem-cell defects underlying abnormal macrophage development and maturation in NOD/Lt mice: defective regulation of cytokine receptors and protein kinase C. AB - The immunopathogenesis of autoimmune insulin-dependent diabetes in NOD mice entails defects in the development of macrophages (M phi s) from hematopoietic precursors. The present study analyzes the cellular and molecular basis underlying our previous finding that the Mo growth factor colony-stimulating factor 1 (CSF-1) promotes a reduced level of promonocyte proliferation and M phi development from NOD bone marrow. CSF-1 stimulation of NOD marrow induced Mos to differentiate to the point that they secreted levels of tumor necrosis factor alpha equivalent to that of controls. However, CSF-1 failed to prime NOD M phi s to completely differentiate in response to gamma-interferon, as shown by their decreased lipopolysaccharide-stimulated interleukin 1 secretion. These defects, in turn, were associated with an inability of CSF-1 to up-regulate c-fms (CSF-1 receptor) and Ifgr (gamma-interferon receptor) expression. Even though the combination of CSF-1 and gamma-interferon up-regulated c-fms and Ifgr transcript levels in NOD M phi s to levels induced in control M phi s by CSF-1 alone, the protein kinase C activities coupled to these receptors remained 4-fold lower in NOD M phi s than in M phi s derived from the marrow of diabetes-resistant NON and SWR control mice. Despite expressing the diabetogenic H-2g7 haplotype, M phi s derived from cytokine-stimulated marrow of the NON.H-2g7 congenic stock were functionally more mature than similarly derived M phi s from NOD mice. Whereas diabetes resistance was abrogated in 67% of irradiated (NOD x NON)F1 females reconstituted with NOD marrow, no recipients became diabetic after reconstitution with a 1:1 mixture of marrow from NOD and the congenic stock. Thus, failure to develop functionally mature monocytes may be of pathogenic significance in NOD mice. PMID- 8415752 TI - Single unit components of the hypothalamic multiunit electrical activity associated with the central signal generator that directs the pulsatile secretion of gonadotropic hormones. AB - Vertebrate reproduction is dependent on the operation of a central signal generator that directs the episodic release of gonadotropin-releasing hormone, a neuropeptide that stimulates secretion of the pituitary gonadotropic hormones and, thereby, controls gonadal function. The electrophysiological correlates of this pulse generator are characterized by abrupt increases in hypothalamic multiunit electrical activity (MUA volleys) invariably associated with the initiation of secretory episodes of luteinizing hormone. Using cluster analysis, we extracted single units from the multiunit signals recorded from the mediobasal hypothalamus of four intact and four ovariectomized rhesus monkeys. Of the 40 individual units identified in this manner, 24 increased their frequency with the MUA volleys. The onset and termination of these single-unit bursts occurred coincidently with those of the MUA volleys in both intact and ovariectomized animals, indicating that the longer duration of the MUA volleys characteristic of the gonadectomized animals was due not to the sequential activation of different units but to the longer bursts of the individual cells. Four other units showed decreases in firing rate during the MUA volleys, while the frequency of the remainder did not change. All the examined units were active during the intervals between the volleys of electrical activity. The results indicate that the MUA volleys associated with the activity of the gonadotropin-releasing hormone pulse generator represent the simultaneous increase in firing rate of some individual hypothalamic neurons and the decrease in the frequency of others. PMID- 8415753 TI - [d-Ala2]deltorphin I-like immunoreactivity in the adult rat brain: immunohistochemical localization. AB - Using a specific antiserum recently raised against [D-Ala2]deltorphin I (DADTI: Tyr-D-Ala-Phe-Asp-Val-Val-Gly-NH2), a highly selective ligand for delta-opioid receptors, we have previously demonstrated the occurrence of positive immunostaining in several structures of mouse brain. We describe here the neuroanatomical distribution patterns of DADTI-immunoreactive neuronal bodies, axons, and tanycytes in rat brain. Positive neuronal somata were localized mainly in the ventral mesencephalon, including the ventral tegmental area and the pars compacta of the substantia nigra. A minor population of positive somata was found in the pars reticulata and pars lateralis of the substantia nigra, raphe nuclei, supramammillary nucleus, and retrorubral reticular nucleus. All these regions, except for the supramammillary nucleus, contain dopamine cell bodies. Intensely stained positive nerve fibers could be traced along the medial forebrain bundle. Dense positive terminals were seen in the neostriatum, nucleus accumbens shell, olfactory tubercle, septal areas, cingulate, and medial prefrontal cortex. Double immunostaining study revealed that, in the substantia nigra, almost all (97.8%) DADTI-positive neurons colocalized with tyrosine hydroxylase (TH), and the doubly stained cells occupied about one-third (29.1%) of the total population of TH positive neurons. Only a few DADTI/TH-positive cells also stained for 28-kDa calbindin D, although many neurons double-stained for 28-kDa calbindin D and TH. In contrast, the supramammillary nucleus contained a number of DADTI-positive cells, which nearly always stained positively for 28-kDa calbindin D but did not stain for TH. The association of DADTI-like immunoreactivity with certain dopaminergic pathways seems of particular interest. A small population of DADTI immunostained tanycytes was present in the ventral part of the third ventricle wall. PMID- 8415754 TI - Integrin overexpression induced by high glucose and by human diabetes: potential pathway to cell dysfunction in diabetic microangiopathy. AB - The nature of the process leading to the acellular nonperfused capillaries of diabetic microangiopathy remains unknown. Because these capillaries manifest thickened basement membranes, we asked whether the process causing deposition of excess extracellular matrix in diabetes modifies cell-matrix interactions in a direction that would compromise cell renewal. In 44 individual isolates of human umbilical vein endothelial cells we observed that high glucose concentrations (30 mM) induce coordinate increases in the levels of mRNAs encoding fibronectin and the fibronectin-specific integrin receptor alpha 5 beta 1 as well as in the cognate proteins. Expression of the integrin subunit alpha 3, component of the alpha 3 beta 1 polyspecific receptor for fibronectin, laminin, and collagen, was also up-regulated by high glucose. Overexpression of integrins correlated with increased cell attachment to exogenous fibronectin and laminin as well as to complex matrix. Moreover, cells exhibited firmer steady-state adhesion to their own matrix. To correlate these in vitro observations with events in human diabetic retinopathy we measured integrin levels in retinal trypsin digests prepared from 10 patients with 8.2 +/- 1.6 (mean +/- SE) years of diabetes and 10 age- and sex-matched nondiabetic controls. Microvessels of diabetic patients showed increased immunostaining for beta 1 integrin (P = 0.025) when compared with control microvessels. These data show that high glucose and diabetes increase integrin expression and thus alter the interaction of vascular endothelial cells with their basement membranes in the direction of firmer cell matrix adhesion. This could compromise the migration and replication critical to the reendothelialization process and contribute to microvascular occlusion. PMID- 8415755 TI - Cocaine as a naturally occurring insecticide. AB - Although cocaine has a fascinating and complex medicinal history in man, its natural function in plants is unknown. The present studies demonstrate that cocaine exerts insecticidal effects at concentrations which occur naturally in coca leaves. Unlike its known action on dopamine reuptake in mammals, cocaine's pesticidal effects are shown to result from a potentiation of insect octopaminergic neurotransmission. Amine-reuptake blockers of other structural classes also exert pesticidal activity with a rank order of potency distinct from that known to affect vertebrate amine transporters. These findings suggest that cocaine functions in plants as a natural insecticide and that octopamine transporters may be useful sites for targeting pesticides with selectivity toward invertebrates. PMID- 8415756 TI - Increased amyloid beta-peptide deposition in cerebral cortex as a consequence of apolipoprotein E genotype in late-onset Alzheimer disease. AB - Amyloid beta-peptide (A beta) deposition in senile plaques and cerebral vessels is a neuropathological feature of Alzheimer disease (AD). We examined the possibility that commonly observed variability in A beta deposition in late-onset AD might be related to apolipoprotein E genotype (APOE gene; the two most common alleles are 3 and 4), since APOE4 is a susceptibility gene for late-onset AD and apolipoprotein E interacts strongly with A beta in vitro. In an autopsy series of brains of late-onset AD patients, we found a strong association of APOE4 allele with increased vascular and plaque A beta deposits. Late-onset AD patients with one or two APOE4 alleles have a distinct neuropathological phenotype compared with patients homozygous for APOE3. PMID- 8415757 TI - Postsynaptic factors in the expression of long-term potentiation (LTP): increased glutamate receptor binding following LTP induction in vivo. AB - Several lines of evidence indicate that LTP in the hippocampus is associated with a change in the properties of postsynaptic glutamate receptors. In the present study, we used quantitative autoradiography to examine the binding properties of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA) and N-methyl-D aspartate subclasses of glutamate receptors in frozen brain sections obtained from rats in which perforant-path LTP was induced in vivo. Induction of LTP resulted in a selective increase in [3H]AMPA binding in those hippocampal subfields receiving perforant-path axons. Increases in [3H]AMPA binding in dentate gyrus (stratum moleculare) were highly correlated with the magnitude of LTP recorded in this structure. Scatchard analyses of [3H]AMPA and 6-cyano-7 nitro-[3H]quinoxaline-2,3-dione (an AMPA receptor antagonist) binding in the dentate gyrus indicated that LTP induction resulted in an increase in the number of AMPA receptor binding sites. No changes in the binding of 3H-labeled N-[1 (thienyl)cyclohexyl]piperidine (an N-methyl-D-aspartate receptor antagonist) were observed in any hippocampal subfield. These results suggest that a modification in postsynaptic AMPA receptors plays a role in the expression of synaptic enhancement following LTP induction in the hippocampus. PMID- 8415758 TI - Cloning and expression of a Kv1.2 class delayed rectifier K+ channel from canine colonic smooth muscle. AB - A cDNA (CSMK1) encoding a delayed rectifier K+ channel of the Kv1.2 class was cloned from canine colonic circular smooth muscle and expressed in Xenopus oocytes. These channels appear to be uniquely expressed in gastrointestinal muscles and may participate in the electrical slow wave activity. Functional expression of CSMK1 in Xenopus oocytes demonstrated a K+ current that activated in a voltage-dependent manner upon depolarization. This current was highly sensitive to 4-aminopyridine (IC50, 74 microM). A low-conductance K+ channel was identified in inside-out patches from oocytes injected with CSMK1. This channel displayed a linear current-voltage relation with a slope conductance of 14 pS. The channels were blocked in a concentration-dependent manner by 4-aminopyridine. Northern blot analysis demonstrated that CSMK1 is expressed in a wide variety of gastrointestinal smooth muscles. Portal vein, renal artery, and uterus do not express CSMK1, suggesting that, among smooth muscles, expression of this K+ channel may be restricted to gastrointestinal smooth muscles. CSMK1 is 91% homologous to RAK, a delayed rectifier K+ channel cloned from rat heart, but displays unique pharmacological properties and tissue distribution. PMID- 8415759 TI - Soft x-ray magnetic circular dichroism: a probe for studying paramagnetic bioinorganic systems. AB - Soft x-ray magnetic circular dichroism was used to study a paramagnetic bioinogranic system. We measured the Fe L edges of Pyrococcus furiosus rubredoxin, using circularly polarized synchrotron radiation, a split-coil super conducting magnet, low sample temperatures, and fluorescence detection. The observed dichroism effect is strong (30%) and in general agreement with the calculation. The method is element- and oxidation state-specific, and the data can be interpreted by established theoretical procedures. Soft x-ray magnetic circular dichroism demonstrates enormous potential as a probe for studying paramagnetic systems in biology, chemistry, and material science. PMID- 8415760 TI - Thermal motions and function of bacteriorhodopsin in purple membranes: effects of temperature and hydration studied by neutron scattering. AB - The internal dynamics of bacteriorhodopsin, the light-driven proton pump in the purple membrane of Halobacterium halobium, has been studied by inelastic neutron scattering for various conditions of temperature and hydration. Light activation can take place when the membrane is vibrating harmonically. The ability of the protein to functionally relax and complete the photocycle initiated by the absorption of a photon, however, is strongly correlated with the onset of low frequency, large-amplitude anharmonic atomic motions in the membrane. For a normally hydrated sample, this occurs at about 230 K, where a dynamical transition from a low-temperature harmonic regime is observed. In moderately dry samples, on the other hand, in which the photocycle is slowed down by several orders of magnitude, no transition is observed and protein motions remain approximately harmonic up to room temperature. These results support the hypothesis, made from previous neutron diffraction studies, that the "softness" of the membrane modulates the function of bacteriorhodopsin by allowing or not allowing large-amplitude motions in the protein. PMID- 8415761 TI - Cloning and properties of the Caenorhabditis elegans TATA-box-binding protein. AB - The nematode Caenorhabditis elegans has become an organism of choice for the study of developmental processes at the genetic level. We have undertaken to develop an in vitro system to study transcription in C. elegans. As a first step we report here the cloning of the cDNA encoding the C. elegans TATA-box-binding protein (CeTBP). We used "touch-down PCR" to generate a specific DNA probe derived from the C-terminal region conserved in all TBP genes cloned to date. Several clones encoding an extended open reading frame were isolated from a phage lambda cDNA library. The complete amino acid sequence of CeTBP deduced from the cDNA reveals a protein of 37 kDa with an extended sequence similarity in the C terminal region with all other TBP cDNAs sequenced so far. The N-terminal region of CeTBP (amino acids 1-153), however, does not show any homology with TBPs from other organisms. Interestingly, the N-terminal portion of the molecule contains three short direct repeats. Purified recombinant CeTBP binds specifically to the TATA box sequence, interacts with transcription factors TFIIA and TFIIB, and is able to substitute for the TFIID basal activity when assayed by in vitro transcription in both HeLa and C. elegans nuclear extracts. CeTBP is therefore a basal transcription factor. PMID- 8415762 TI - Reconstruction of neural tube-like structures in vitro from primary neural precursor cells. AB - Vertebrate central nervous system develops from a neural tube derived from the embryonic ectoderm. In mouse, the neural tube around embryonic day 10 primarily consists of neural precursor cells (NPCs). During the development of embryonic central nervous system, NPCs proliferate and migrate outward; thus later stages show NPCs toward the lumen of the neural tube and neurofilament-positive differentiated cells toward the periphery. In conventional liquid culture, NPCs isolated from mouse on embryonic day 10 proliferate and differentiate into neurofilament-positive neurons. In the present communication, we show that fragments of neural tubes and aggregates of NPCs, when placed into collagen gel matrix, form three-dimensional structures which resemble the neural tube formed in vivo in the developing embryos. Even dissociated NPCs form the three dimensional structures in the collagen gel matrix. Our results indicate that individual NPCs or fragments of neural tubes carry morphogenetic information which allows them to reconstruct neural tube-like structures in vitro. PMID- 8415763 TI - Pheromone action regulates G-protein alpha-subunit myristoylation in the yeast Saccharomyces cerevisiae. AB - Myristic acid (C14:0) is added to the N-terminal glycine residue of the alpha subunits of certain receptor-coupled guanine nucleotide-binding regulatory proteins (G proteins). The G alpha subunit (GPA1 gene product) coupled to yeast pheromone receptors exists as a pool of both myristoylated and unmyristolyated species. After treatment of MATa cells with alpha factor, the myristoylated form of Gpa1p increases dramatically, and the unmyristoylated form decreases concomitantly. This pheromone-stimulated shift depends on the function of STE2 (alpha-factor receptor), STE11 (a protein kinase in the response pathway), and NMT1 (myristoyl-CoA:protein N-myristoyltransferase) genes and uses the existing pool of fatty acids (is not blocked by cerulenin). Myristoylated Gpa1p persists long after pheromone is removed. Because myristoylation is essential for proper G alpha-G beta gamma association and receptor coupling, pheromone-dependent stimulation of Gpa1p myristoylation may be an important contributing factor in adaptation after signal transmission. PMID- 8415764 TI - The hobo transposable element of Drosophila can be cross-mobilized in houseflies and excises like the Ac element of maize. AB - The hobo transposable element from Drosophila melanogaster was found to be capable of excision, resulting in donor sites unlike those reported for any other transposable element currently known in animals. These empty sites most closely resemble those left by the transposable elements Ac and Tam3 in Zea mays and Antirrhinum majus, respectively. Like Ac and Tam3, the hobo element was found to function in a distantly related species, in this case the housefly Musca domestica. Hobo excision in M. domestica was found not to require the expression of hobo-encoded transposase but instead appears to be driven by an endogenous hobo transposase-like activity. DNA sequences displaying high homology to the hobo transposase gene were isolated from the genome of M. domestica and appear to be part of a mobile-element system related to hobo, Ac, and Tam3. PMID- 8415765 TI - The segment of invariant chain that is critical for association with major histocompatibility complex class II molecules contains the sequence of a peptide eluted from class II polypeptides. AB - Major histocompatibility complex class II molecules present peptides from an extracellular source of antigens to CD4+ T lymphocytes. The class II-associated invariant chain affects this role of alpha and beta polypeptides by restriction of peptide loading to endocytic vesicles. Up to now no specific portion of the invariant chain has been defined as the class II binding site. We constructed recombinant invariant chain genes and inspected association of the mutant invariant chains with class II polypeptides. Here we demonstrate that an extracytoplasmic sequence of the invariant chain (aa 81-109) that is only 23 residues away from the transmembrane region is essential for contact with class II polypeptides, whereas the remaining C-terminal part is dispensable for binding. The sequence of invariant-chain-derived peptides that were eluted from class II molecules is contained in this segment and may define the class II binding site of the invariant chain. The membrane-proximal position of this region suggests that the invariant chain and invariant-chain-derived peptides isolated from class II molecules bind to a domain distinct from the class II pocket. PMID- 8415766 TI - Molecular dissection of the mouse interleukin-4 promoter. AB - Understanding the molecular mechanisms regulating the expression of interleukin 4 (IL-4) may shed light on the differentiation of lymphokine-producing phenotypes of CD4+ T cells. We have identified two DNA segments that are necessary for full phorbol 12-myristate 13-acetate (PMA)-induced activity of the IL-4 promoter region in the thymoma cell line EL4. Through deletion and mutation analyses, one of these segments (-57 through -47) was shown to be indispensable for promoter function. We designated this sequence consensus sequence 1 (CS1), as it shares homology with a sequence (ATTTTCCNNTG) that appears five times in the proximal 302-base-pair (bp) region 5' of the gene. We examined CS1 in further detail, as well as a second consensus sequence, CS2, located at nucleotides -75 through -65; both are within a minimal 83-bp construct that expresses full promoter activity. CS1- and CS2-spanning oligonucleotides bound apparently distinct PMA-inducible, sequence-specific factors in mobility-shift assays. Multimer constructs linking CS1- or CS2-spanning oligonucleotides to a heterologous promotor revealed that the CS1 construct had the greater enhancer activity in EL4 cells. Mutating the CS1 sequence within the context of the 302-bp promoter abolished all activity of the promoter, while mutating the CS2 sequence alone had little effect. Furthermore, a CS1 multimer could drive a heterologous promoter in an IL-4 producing [helper T-cell type 2 (TH2-type)] T-cell clone but not in a non-IL-4 producing (TH1-type) clone, suggesting a mechanism by which IL-4 production could be differentially regulated in TH subsets. PMID- 8415767 TI - Ambient glucose and aldose reductase-induced myo-inositol depletion modulate basal and carbachol-stimulated inositol phospholipid metabolism and diacylglycerol accumulation in human retinal pigment epithelial cells in culture. AB - Physiological hyperglycemia has been speculated to alter phosphoinositide (PPI; inositol phospholipid) signal transduction in cells prone to diabetic complications by two separate mass-action mechanisms with antiparallel putative effects on diacylglycerol (DAG): (i) sorbitol-induced depletion of myo-inositol leads to diminished PPI synthesis and turnover and DAG release, and (ii) elevated glucose-derived DAG precursors enhance de novo DAG synthesis. Because the first mechanism is mediated by aldose reductase (AR2), which converts glucose to sorbitol, the effects of glucose on basal and stimulated PPI signaling were explored in lines of cultured human retinal pigment epithelial cells differing widely in their basal AR2 gene expression and enzymatic activity. The results suggest that the effects of glucose on PPI signaling vary inversely with the level of AR2 activity and parallel the extent of AR2-induced myo-inositol depletion. PMID- 8415768 TI - A gene affecting Wallerian nerve degeneration maps distally on mouse chromosome 4. AB - When a nerve axon is cut or crushed, the nerve fibers in the distal part of the axon, separated from the cell body, undergo a form of spontaneous degeneration, known as Wallerian degeneration. A substrain of the mouse inbred strain C57BL, known as C57BL/Ola, carries a mutant form of a gene involved in Wallerian degeneration in the peripheral and central nervous systems, and in retrograde degeneration of retinal ganglion cells. Wallerian degeneration in this substrain is abnormally slow. Previously the defect had been shown to be due to an autosomal dominant gene. The locus has been given the name and symbol Wallerian degeneration Wld, with the mutant allele Wlds (Wallerian degeneration-slow). The Wld locus has now been mapped, by using conventional and molecular markers, to the distal end of chromosome 4, near the locus of pronatriodilatin (Pnd). The order of loci (with recombination distances in centimorgans, cM) is cen-D4Mit11 8.9 +/- 1.7 cM-Fuca-2.5 +/- 0.93 cM-Akp-2-3.2 +/- 1.1 cM-D4Mit48-3.5 +/- 1.1 cM (Wld, Pnd, D4Mit49)-0.71 +/- 0.50 cM-(Eno-1, D4Mit33)-1.4 +/- 0.70 cM-D4Mit42-2.5 +/- 0.93 cM-D4Smh6b. The information on the position of the Wld locus should be valuable in further characterization of this gene involved in nerve degeneration and regeneration. PMID- 8415769 TI - Effects of exercise on plasma lipids and lipoproteins of women. AB - This review summarizes the cross-sectional and training studies (acute and chronic) that have examined the relationship between exercise and plasma lipid and lipoproteins in women. Because women experience major fluctuations in reproductive hormones throughout the life cycle, the effects of the endogenous sex steroid status on the association between exercise and plasma lipoproteins also are addressed. In general, cross-sectional studies report a positive association between exercise and high density lipoprotein-cholesterol (HDL-C) in both pre- and postmenopausal women. Women on hormone replacement therapy who report exercising have higher HDL-C than sedentary women on hormone replacement therapy. Results from longitudinal training studies have been inconsistent because of experimental design, i.e., inadequate type, duration, and intensity of exercise intervention, lipid measurements made across the menstrual cycle, and studies carried out in women with high baseline HDL-C. Since lipids vary approximately 10-25% throughout the menstrual cycle, menstrual phase should be controlled when determining lipid changes after an exercise intervention. In approximately half of the intervention studies, an increase in HDL-C was demonstrated; the magnitude of the response that can be expected is approximately 10%. The responsiveness of pre- versus postmenopausal women to an exercise intervention is unknown. Studies are needed to clarify the interactive effects of exercise and sex hormones on plasma lipoproteins in women of all ages. This information will be useful in developing intervention programs to reduce the risk of coronary heart disease in women. PMID- 8415770 TI - Orally administered interferons suppress bone marrow function. AB - The accepted routes of interferon (IFN) administration in clinical applications are intramuscular, subcutaneous, intraperitoneal, intratumor, and intravenous. Recently, oral administration of interferons has been shown to cause a suppression of peripheral white blood cell (WBC) counts. Moreover, orally administered interferons mediate their peripheral WBC suppression via a different mechanism than that of intraperitoneally administered interferons. This study extends the previous studies to show that the peripheral WBC suppression induced by oral interferon treatment reflects an actual bone marrow suppression. The bone marrow-suppressive effects of orally and subcutaneously administered recombinant human IFN-alpha A/D (rHuIFN-alpha A/D) have been partially characterized in kinetics studies and compared with the peripheral WBC-suppressive effects of orally and subcutaneously administered rHuIFN-alpha A/D. Oral and subcutaneous administrations of rHuIFN-alpha A/D cause a significant suppression of peripheral WBC counts with 1 day of rHuIFN-alpha A/D administration. This suppression reaches its maximum level with 3 days of rHuIFN-alpha A/D administration and plateaus over a 12-day treatment time. Similarly, oral and subcutaneous administrations of rHuIFN-alpha A/D cause a significant suppression of bone marrow function with 1 day of rHuIFN-alpha A/D administration. This suppression reaches its maximum level with 3 days of rHuIFN-alpha A/D administration and plateaus over a 12-day treatment time. Thus, the WBC-suppressive and bone marrow suppressive effects of rHuIFN-alpha A/D administered either orally or subcutaneously parallel each other. The peripheral WBC-suppressive activities of orally and subcutaneously administered rHuIFN-alpha A/D diminish at the same rate, after cessation of rHuIFN-alpha A/D treatment. Peripheral WBC suppression is lost by 5 days after cessation of rHuIFN-alpha A/D treatment. The mechanisms by which orally and subcutaneously administered interferons exert their bone marrow-suppressive effects differ, however. Bone marrow suppression mediated by subcutaneous administration of murine IFN-alpha/beta (MuIFN-alpha/beta) is blocked by the presence of circulating antibodies to MuIFN-alpha/beta. In contrast, the bone marrow suppression mediated by oral administration of MuIFN alpha/beta occurs even in the presence of circulating antibodies to MuIFN alpha/beta. These results continue to support a potential clinical role for oral administration of interferons, particularly for the control of diseases of myelogenous origin. PMID- 8415771 TI - Intestinal mucosal ornithine decarboxylase and brush border membrane vesicle Na(+)-H+ exchange activities in diabetic rats. AB - To determine the possibility that intestinal mucosal ornithine decarboxylase activity can modulate mucosal brush border membrane Na(+)-H+ exchange activity, we studied the relationship between jejunal mucosal ornithine decarboxylase activity and mucosal brush border membrane Na(+)-H+ exchange activity in adolescent streptozotocin-diabetic and normal control rats. Diabetes was associated with enhanced intestinal mucosal ornithine decarboxylase and Na(+)-H+ exchange activities. Groups of diabetic and control rats were given difluoromethylornithine in drinking water to suppress intestinal mucosal ornithine decarboxylase activity. As expected, 10 days after induction of diabetes, intestinal mucosal weight (67.7 mg/cm vs 56.1 mg/cm), DNA (47.3 micrograms/mg protein vs 32.7 micrograms/mg protein), ornithine decarboxylase activity (1107 units/hr vs 654 units/hr), and brush border membrane vesicle Na(+) H+ exchange activity, assessed as Vmax of 22Na+ uptake (32.5 nmol/mg protein/15 min vs 15.2 nmol/mg protein/15 min), were significantly greater in diabetic than in control rats. Treating diabetic and control rats with difluoromethylornithine suppressed jejunal mucosal growth by over 30%, ornithine decarboxylase activity by over 80%, and brush border membrane vesicle 22Na+ uptake by over 60%. Highly significant direct correlations (r > 0.900) were observed between jejunal DNA content, mucosal ornithine decarboxylase activity, and brush border membrane vesicle Na(+)-H+ exchange activity. The above findings suggest that jejunal mucosal ornithine decarboxylase activity can modulate mucosal epithelial proliferation and mucosal brush border membrane Na(+)-H+ exchange activity. PMID- 8415772 TI - Drugs that suppress hepatic fat synthesis in starved-refed BHE/cdb rats also have an effect on muscle protein synthesis. AB - Simultaneous lipogenesis and protein synthesis as influenced by LY79771, testosterone, or dehydroepiandrosterone in starved/refed rats were studied. Starved-refed BHE/cdb rats were injected with one of these compounds during the 2 day refeed period. Hepatic de novo fatty acid synthesis using tritium incorporation into fatty acids and protein synthesis using [14C]phenylalanine incorporation into hepatic and muscle protein were determined. Hepatic lipogenesis was decreased by all three drugs and these drugs had a differential effect on protein synthesis. We did not observe a corresponding increase in protein synthesis in the liver when fat synthesis was decreased, but we did observe a corresponding increase in muscle protein synthesis. We concluded that in the acute hyperlipogenic state induced by starvation/refeeding, these drugs induced a reciprocal increase in muscle protein synthesis along with a suppression of fatty acid synthesis. PMID- 8415773 TI - Age-related response to dietary fructose in the rat: discrepancy in triglyceride and apolipoprotein B synthesis as a possible mechanism for fatty liver induction in adult rats. AB - The effects of fructose feeding on plasma and liver lipids, triglyceride secretion, and plasma apolipoprotein B and their liver mRNA level were studied in young and adult rats. We have shown that the responsiveness of adult rats to dietary fructose differs from that of young rats with regard to body parameters as well biochemical analyses. In young rats, fructose diet causes a coupled induction of liver triglyceride and apolipoprotein B synthesis via increased mRNA level. In adult rats it appears that triglyceride secretion is lower and less inducible by dietary fructose than in young rats. This insufficient export of the excess of synthesized triglycerides may cause fatty liver in adult animals. Reduced adaptation of liver lipoprotein secretion to dietary carbohydrates in adult animals may be explained by the failure to stimulate apolipoprotein B synthesis at the mRNA level in these rats. PMID- 8415774 TI - Plasma corticosterone response to chronic ethanol consumption and exercise stress. AB - Acute exposure to ethanol induces a stress response in mice that is manifested by increased plasma corticosterone (PC) concentration. However, during chronic intake of 7.5% w/v ethanol, diurnal fluctuation of PC is dampened. Whether chronic consumption of 20% w/v ethanol alters normal diurnal fluctuation of plasma glucocorticoids is not known. Investigating the PC response in 20% w/v ethanol-consuming mice is of interest because glucocorticoids are known suppressants of natural killer (NK) cell activity and increased concentration or altered diurnal fluctuation of PC may have a modulatory role on NK cells in these mice. Mice given 20% w/v ethanol for at least 7 days and for as long as 10 weeks have suppressed splenic NK cell cytolytic activity. Thus, the purpose of this study was to examine whether mice given 20% w/v ethanol exhibited normal concentrations and diurnal variation of PC. To further define the glucocorticoid response in chronic ethanol-consuming mice, PC concentration was evaluated in response to a secondary stress of physical exercise. After 1 week, ethanol consuming mice exhibited abnormal diurnal PC periodicity that was progressively dampened during the remaining 9 weeks. Acute physical exercise during Week 1 induced a 2-fold increase in PC concentration compared with pre-exercise values, a response that was independent of ethanol intake. After 6 and 10 weeks, the postexercise PC concentration was attenuated in ethanol-consuming compared with water-drinking mice. It was concluded that suppressed NK cell activity typically observed with this model of chronic ethanol intake is not directly associated with dampened diurnal fluctuation in PC. PMID- 8415775 TI - Gonadotropin hormone-releasing hormone induced luteinizing hormone responses in young and old female C57BL/6J mice. AB - The estrogen-induced luteinizing hormone (LH) surge is markedly attenuated in old (24 months) female mice compared with young (6 months) females. To test whether or not this attenuated LH response is due to a diminished capacity of the pituitary to respond to hypothalamic gonadotropin hormone-releasing hormone (GnRH), the pituitary response to exogenous GnRH was measured in young (5-6 months), normally cycling (n = 6) and old (24 months), acyclic constant diestrus (n = 6) C57BL/6J female mice. Mice were ovariectomized and estrogen-treated for 7 days. After intrajugular catheterization on Day 6, serial blood samples were taken at 15-min intervals for 165 min on Day 7. Serum LH was measured in samples obtained before and after infusion of either saline or GnRH (5 micrograms/5 microliters saline/kg body wt) and 15 micrograms/15 microliters saline/kg body wt 1 hr apart). Saline-treated animals demonstrated no LH response in either young (0.09 +/- 0.02 ng/ml baseline) or old (0.11 +/- 0.01 ng/ml baseline) females. However, a significant release of LH was obtained in response to each challenge of GnRH in both young (0.3 +/- 0.04 ng/ml first challenge, 0.69 +/- 0.1 ng/ml second challenge) and old (0.78 +/- 0.1 ng/ml first challenge, 1.76 +/- 0.2 ng/ml second challenge) mice. The LH response in the aged group was significantly greater (P < 0.05, analysis of variance) than in the young group. These results show that pituitaries of old female mice were at least as capable of responding to exogenous stimulation by GnRH as those of young. We conclude that alteration in the capacity of the pituitary to respond to GnRH is not likely to be a factor contributing to altered LH secretion with age in C57BL/6J mice. PMID- 8415776 TI - Stimulation of phosphoinositides by agents that stimulate proton secretion in toad urinary bladder. AB - The urinary bladder of Bufo marinus excretes H+ and this excretion is increased by metabolic acidosis (MA), insulin (IN), prostaglandin E2 (PGE2), increases CO2, and aldosterone. The purpose of this experiment was to determine whether MA, IN, PGE2, CO2, and aldosterone stimulate inositol phosphate's (IP) formation in isolated cells of toad urinary bladder. Cells were prepared by treating bladder sacs with collagenase. Cells were obtained from 10 toads in MA and 10 normal toads, suspended in 2 ml of Ringer's solution containing LiCl (10 mM), myo inositol (5 mM), and [3H]myo-inositol (10 microCi), and then incubated for 2 hr at 25 degrees C. Cells were homogenized and the IP fractions quantitated by column chromatography and liquid scintillation counting. The results were expressed as dpm (mu MPO4)-1 (hr)-1. The IP in MA cells was 44,202 +/- 4,646 and in normal toad cells it was 31,637 +/- 3,613 (P < 0.05). In a separate experiment, cells from 10 paired hemibladders were isolated from normal toads. The cells were treated exactly as above except there were no LiCl in the bath. LiCl was added to all baths after 2 hr and the experimental cells were challenged with IN, PGE2, increases CO2, and aldosterone for 20 min. The IP were quantitated as above. IN treatment stimulated inositol bisphosphate and inositol triphosphate (P < 0.01). PGE2 and increases CO2 also stimulated inositol triphosphate (P < 0.05). Aldosterone did not alter formation of any of the IP fractions. We conclude that MA, IN, PGE2, and increases CO2 stimulate IP formation in cells of toad urinary bladder and inositol triphosphate may be an important second messenger in mediating the response of MA, IN, PGE2, and increases CO2. PMID- 8415777 TI - Chronic effects of camostate on growth and endocrine function of the pancreas in streptozotocin-induced diabetic rats. AB - Chronic feeding of normal rats with camostate, a synthetic trypsin inhibitor, stimulates the growth of the pancreas chiefly by increasing cholecystokinin release. We examined the effects of camostate on the growth and the endocrine function of the pancreas in streptozotocin-induced diabetic rats. The pancreatic weight of the diabetic rats given camostate (200 mg/kg/day) by oral gavage for 14 days was significantly elevated by 120% over that of diabetic rats not given camostate, and was comparable to that in the nondiabetic rats given camostate. The total pancreatic contents of DNA, RNA, and protein in diabetic rats given camostate were also significantly higher than those in diabetic rats not given camostate, and did not differ from those observed in nondiabetic rats given camostate. The pancreatic growth seen in diabetic rats treated with camostate was associated with moderate hyperplasia and pronounced hypertrophy. By contrast, treatment with camostate did not improve hyperglycemia, hypoinsulinemia, or low pancreatic content of insulin seen in diabetic rats. These findings demonstrate a marked growth of the pancreas in diabetic rats stimulated by camostate, and suggest that camostate-induced pancreatic growth is not affected by the reduced level of the endogenous insulin. The present study also indicates that camostate has no beneficial effects on the function of residual B cells, failing to improve diabetes. PMID- 8415778 TI - Transcobalamin II mediated delivery of albumin-bound hydroxocobalamin to human liver cells. AB - We show that hydroxocobalamin bound to human serum albumin can dissociate and bind to transcobalamin II present in serum. Human liver cells in culture exposed to hydroxocobalamin bound to albumin incorporated less of the vitamin than when similar amounts of unbound hydroxocobalamin or cyanocobalamin were present. In the presence of transcobalamin II, a 4.5-fold increase in cellular uptake occurred, but this amount was less than when hydroxocobalamin or cyanocobalamin were added to transcobalamin II. These results indicate that albumin, by binding hydroxocobalamin, can alter the dynamics of binding to transcobalamin II and the subsequent cellular incorporation of this form of the vitamin. PMID- 8415779 TI - Involvement of cAMP and protein kinase C in cytomegalovirus enhancement of human immunodeficiency virus replication. AB - Cytomegalovirus (CMV) infection constitutes a serious threat to patients with acquired immune deficiency syndrome. Recently we reported that human immunodeficiency virus (HIV) infection of CD4+ cells was associated with sustained elevation of cellular levels of cAMP. Moreover, cyclic nucleotide modulators enhanced HIV replication by increasing intracellular levels of cAMP. In this study, the effect of CMV on HIV replication in CMV/HIV mixed infection and its relationship to cAMP were examined. MT-4 cells, CMV strain AD169, and HIV strain IIIB were used. Optimal enhancement (4.4-fold increase) of HIV replication was observed when MT-4 cells were infected with CMV at Day 0 followed by HIV on Day 4 after infection, as determined by reverse transcriptase activity on Day 11 after infection. cAMP (measured by radioimmunoassay) levels in cells infected with CMV alone, HIV alone, or CMV/HIV together were 2-, 3-, and 5-fold above untreated cells, respectively. CMV also enhanced the replication of UV-irradiated HIV 4-fold and this was associated with a 2-fold increase in cAMP as well. Moreover, UV-irradiated CMV enhanced HIV replication 8.8-fold. The same dose of viable and UV-irradiated CMV used in the above experiments increased protein kinase C activity in these cells 3.0- and 8.0-fold, respectively. These findings might suggest that cAMP and protein kinase C are involved in CMV enhancement of HIV replication. These findings may have relevance to the identification of novel target sites for development of antiviral therapeutics. PMID- 8415780 TI - Influence of prolactin and growth hormone on the activation of dwarf mouse lymphocytes in vivo. AB - The influence of recombinant bovine prolactin (PRL) and recombinant bovine growth hormone (GH) was examined on the popliteal lymph node (PLN) expression of interleukin-2 receptors (IL-2R) in female Snell dwarf mice and normal litter mates after concanavalin A footpad injection. The absolute number of PLN CD4+, CD8+, or B+ cells of dwarf mice was less than that observed for normal litter mates, but when adjusted for the difference in body weight, only the absolute number of B cells was lower in dwarf animals when compared with normal litter mates. The injection of PRL or GH did not alter the observation. The administration of recombinant bovine PRL to normal animals, but not recombinant bovine GH, increased the expression of IL-2R or unstimulated PLN CD4+ and CD8+ subsets. Hormone administration to dwarf animals, however, did not alter the expression of IL-2R on unstimulated PLN T cell subsets. PLN cells from dwarf animals were poorly activated in vivo after injection of concanavalin A and the level of IL-2R expression induced was only 50% of that seen in the PLN of normal animals. The administration of PRL and GH completely corrected the defective induction of IL-2R expression on PLN from dwarf animals after concanavalin A stimulation. These findings strongly suggest that PRL and/or GH play an important role at some stage of the T cell activation process in vivo. Further studies are needed to precisely identify the defect in the dwarf mice. PMID- 8415781 TI - Effect of immobilization stress on plasma luteinizing hormone, testosterone, and corticosterone concentrations and on 3 beta-hydroxysteroid dehydrogenase activity in the testes of adult rats. AB - We have examined the effect of 3 hr of immobilization stress on plasma luteinizing hormone, testosterone, and corticosterone levels, and on the activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) in microsomal and mitochondrial fractions of the testis from adult rats. Immobilization for 3 hr increased plasma corticosterone and reduced plasma testosterone concentrations by 57%. Plasma luteinizing hormone levels were lower, although not significantly (P = 0.093) so, in stressed animals. Immobilization (3 hr) reduced the Vmax values of 3 beta-HSD in the mitochondria and in the microsomal fraction of the testis by 40% and 34%, respectively, but had no effect on the Km values of 3 beta-HSD in the two cellular compartments. These results suggest that the inhibition of the activity of 3 beta-HSD may be partially responsible for the disruption of testicular steroidogenesis during immobilization stress. PMID- 8415782 TI - Contracts for improvement. A contractual basis for implementing nursing audit. AB - 1. Explicit contracts between partners in care are needed for the effective audit of nursing. 2. Questioning the validity and reliability of audit tools can enhance their use. 3. The pain of change should be seen to be shared by all disciplines. 4. Quality in nursing is a dynamic and developing concept. PMID- 8415783 TI - An overview of current research into diabetes. AB - 1. Current research into diabetes will influence a decade of change in care and treatment. 2. Several new oral drugs are currently under development to treat non insulin dependent diabetes. 3. Clinical trials are underway into new routes to deliver insulin therapy. PMID- 8415784 TI - Cutting the delay reduces the risk. Assessment of the risk of developing pressure sores among elderly patients in A&E. AB - 1. A fractured neck of femur is one of the most common presentations in elderly people within A&E departments, and they have a high incidence rate of pressure sores. 2. Patients should have their pressure sore risk calculated on admission and a triage category assigned. 3. A district approach to pressure sore prevention and equipment should be undertaken by health authorities. 4. Preventive measures should be utilised within A&E departments to reduce the risk of pressure sore development. PMID- 8415785 TI - More information equates with less anxiety. Reducing anxiety in cervical screening. AB - 1. Anxiety may be a factor influencing attendance for cervical screening. 2. Anxiety may be caused by stress and embarrassment of the test procedure and/or misunderstanding its purpose. 3. Women who were given the routine procedure were compared with a study group who were given an extra 10 minutes discussion by nurses. 4. The study group rated the explanation as being clearer, and after receiving the result they reported significantly less anxiety than those in the routine group. PMID- 8415786 TI - Professional development through distance learning. The Professional Nurse Accredited Learning Scheme. AB - 1. Professional Nurse Accredited Learning Scheme enables nurses to achieve CATS points towards a diploma in higher education. 2. The scheme also helps nurses fulfil their PREP requirements. 3. Each module in the scheme is presented in four parts over four months. 4. Modules are based on specific learning outcomes, tested through a two-part written assessment. 5. Nurses can undertake only part one of the assessment and gain 7.5 level 2 credits, both parts of the assessment and gain 15 level 2 credits, or they can choose not to be assessed and receive no academic credits. PMID- 8415787 TI - When savings are costly for patients. PMID- 8415788 TI - Management of asthma in pregnancy. AB - 1. Approximately a third of mothers find their asthma improves during pregnancy, while one third stay the same and the remainder's asthma worsens. 2. Asthma needs to be controlled; if it is not, the fetus may receive insufficient oxygen. 3. The two main types of asthma arise from different causes and have different patterns of inheritance, although in many cases the originating stimulus is the same. 4. A polluted environment contributes to the production of allergens and irritants which trigger asthma in susceptible people. PMID- 8415789 TI - Access to the coping strategies. Managing anxiety in elective surgical patients. AB - 1. The prospect of surgery increases anxiety and reduces cognitive thinking, reducing the ability to cope. 2. Patients use information to reduce their anxiety. 3. Preoperative information has been shown to be effective in improving patient outcomes after surgery. 4. Theatre nurses must base their practice on research, and embrace a perioperative role, using such interventions as preoperative teaching, preparatory sensory information and active listening. PMID- 8415790 TI - Homoeopathy: an energy level therapy. AB - 1. Homoeopathy is still in its infancy as a developing scientific and effective medical system. 2. Homoeopathy is an energy level therapy, not a chemical level therapy. 3. The homoeopathic philosophy and treatment of illness can be considered to be diametrically opposed to orthodox western medicine's philosophy and treatment. 4. People consulting homoeopaths should ensure they are licensed from a college, or registered with The Society of Homoeopaths. PMID- 8415791 TI - A strategy to enhance skills. Developing intravenous therapy skills for community nursing. AB - 1. Information abounds on teaching clients, partners and relatives about home i.v. therapy, but there is a lack of training for community nurses in this area. 2. Training for IV therapy has been difficult to develop in community settings, but must involve managers, specialists, and practitioners. 3. Community-led IV therapy training requires resources across acute and community sectors, with skills development through contract learning and focusing skills to specific practice contexts. 4. Training and experience in managing central venous access developed by community nurses are transferable to all clients and to other colleagues. PMID- 8415793 TI - Preventing and managing work-related strain injuries. PMID- 8415792 TI - Take the strain out of repetitive movement. Management of tenosynovitis and upper limb disorders. AB - 1. Rapid repetitive movements in connection with machine-pacing can lead to the development of upper limb disorders and repetitive strain injuries. 2. The main treatment of these disorders is rest and physiotherapy. 3. Operating VDUs is not a high risk occupation, but can lead to work-related upper limb disorders. 4. Occupational health nurses have a major part to play in preventing repetitive strain injuries in industry. PMID- 8415794 TI - What is a nursing audit facilitator? PMID- 8415795 TI - Animal models of human lipid metabolism. PMID- 8415796 TI - Trans monounsaturated fatty acids in nutrition and their impact on serum lipoprotein levels in man. AB - Trans-C18:1 in the diet originate predominantly from partially hydrogenated oils, with beef, mutton and dairy products being an additional source. These fatty acids are absorbed and incorporated into lipids. Their estimated consumption is about 5-7% of total fatty acids, although reliable data are lacking. In addition, large variations between individuals exist. There is no evidence that trans fatty acids accumulate in human tissues. Elaidic acid and its positional isomers do, however, raise LDL cholesterol and apoprotein B and Lp(a) and probably depress HDL cholesterol and apoprotein A-I, compared with the cis isomer, oleic acid. In view of these adverse effects, patients at high risk for atherosclerosis, in addition to reducing their intake of saturated fatty acids and of cholesterol might also do well to avoid excessive intakes of trans fatty acids. Still, trans fatty acids form only a minor component of the diets of most patients and therefore even marked relative reductions in intake will probably have less of an impact on LDL cholesterol than a sizeable reduction in saturated fatty acids and cholesterol will produce. PMID- 8415797 TI - Sphingolipids in lower animals. PMID- 8415798 TI - Phospholipid biosynthesis in protozoa. AB - Because of the diverse nature of the organisms which are all classed as 'protozoa' (and because of the lack of detailed information on phospholipid metabolism about most of them), it will probably never be possible to generalize phospholipid metabolism to the degree that it has been possible to characterize a mammalian metabolism. Nonetheless, patterns have begun to emerge (i.e. the similarities among the ciliates Entodinium, Paramecium and Tetrahymena) and will not doubt be expanded upon in the future. PMID- 8415799 TI - Hyperlipidaemia and hypercoagulability. PMID- 8415800 TI - Free radical-lipid interactions and their pathological consequences. AB - In this review we have tried to present the current thinking on the consequences for lipids of their interactions with free radicals and the pathological implications. In particular, atherosclerosis and cancer have been addressed. In the case of the former, it is not clear whether the initial oxidative event is an enzymic or free radical-mediated process as yet. However, the importance of the antioxidants in controlling LDL oxidation, macrophage uptake of oxidatively modified LDL and progression of atheroma in animal models certainly suggests an important propagative role for free radical-mediated events. With regard to cancer, oxidative modification of cell lipids has potential consequences for tumour cell proliferation. Whilst lipid hydroperoxides can serve as an origin of prostaglandins with tumour inhibitor (or immunosuppressive) properties, they may also influence cellular growth regulatory proteins normally dependent on membrane lipid integrity. Alternatively, they may function as a source of aldehydic breakdown products capable of 'down-regulating' cell proliferation through covalent modification of regulatory proteins. Oils rich in n-3 polyunsaturated fatty acids have toxic effects towards tumour cells. This toxicity is not mediated by prostaglandins but rather through the capacity of such agents to elevate the levels of lipid peroxides. This may be enhanced by active oxygen species released constitutively from tumour cells. PMID- 8415801 TI - Potential antitumor agents. XX (1). 6-Anilinoimidazo[2,1-b]thiazoles. AB - The synthesis of 6-anilinoimidazo[2,1-b]thiazoles, related to the well-known antitumor agent amsacrine, is reported. The cytotoxic activity of the new compounds was evaluated on HeLa cells. Compound 3a, the most closely related to amsacrine, was significantly active. PMID- 8415802 TI - Monthly prostaglandin bibliography prepared by the University of Sheffield Biomedical Information Service. PMID- 8415803 TI - Migraine: implication of arachidonic acid metabolites. PMID- 8415804 TI - Effect of 5-lipoxygenase and cyclooxygenase blockade on porcine hemodynamics during continuous infusion of platelet-activating factor. AB - We hypothesized that 5-lipoxygenase and cyclooxygenase products might be mediators of cardiopulmonary and systemic vascular effects induced by a 4 h continuous infusion of platelet-activating factor (PAF, 10 ng/kg/min) in anesthetized pigs. Indomethacin (cyclooxygenase inhibitor) potentiated and CGS 8515 (5-lipoxygenase inhibitor) attenuated PAF-induced increases in total peripheral resistance (TPR) from 2.5 to 4 h. However, the 5-lipoxygenase inhibitor failed to modify pulmonary vasoconstriction and hypertension caused by PAF. Except for a delay in onset (approximately 44 s) and rate of development of pulmonary hypertension during the initial 10 min of PAF infusion, the pulmonary hemodynamic changes were also not attenuated by indomethacin. On the other hand, at 4 h, the PAF-induced pulmonary hypertension and systemic vasoconstriction were completely or partially reversed, respectively, by WEB 2086 (PAF receptor antagonist). The PAF-induced increases in plasma thromboxane B2 (TXB2) were blocked by indomethacin but not by CGS 8515, and at 4 h the 5-lipoxygenase inhibitor potentiated the levels of TXB2 in pigs treated with PAF. The plasma concentrations of 6-keto-PGF1 alpha and leukotriene B4 (LTB4) were not modified by PAF or CGS 8515 + PAF. We conclude that PAF-induced increases in TPR (2.5-4 h) are potentiated by indomethacin and are dependent on 5-lipoxygenase products other than LTB4. Although the early pulmonary vascular response (< 10 min) to PAF is dependent on cyclooxygenase products, the sustained response (after 10 min) cannot be explained by either 5-lipoxygenase or cyclooxygenase products but may be mediated directly by PAF receptors. PMID- 8415805 TI - Dietary fish oil supplementation alters LTB4:LTB5 ratios but does not affect the expression of acute graft versus host disease in mice. AB - One of the mechanisms by which corticosteroids may modify acute graft vs host disease (GvHD) is via inhibition of arachidonic acid (AA) metabolism. Leukotriene B4 (LTB4) is a product of that pathway which may take part in the pathogenesis of GvHD through the stimulation of T-lymphopoiesis and T-lymphocyte activation. LTB4 is a metabolite of AA (20:4n-6). Alternate dietary sources of polyunsaturated fatty acids (PUFA), specifically eicosapenteinoic acid (20:5n-3) (EPA) and docosahexaenoic acid (22:6n-3) (DHA), shift the LTs formed with a decrease in LTB4 an increase in LTB5. LTB5 is a less potent agonist than LTB4 and this results in a theoretical decrease of LTB4 mediated events. Supplementation of in vitro bone marrow cultures with EPA or DHA had no detrimental effect on myeloid colony formation. Dietary EPA/DHA supplementation in mice with induced GvHD appeared to be safe and well tolerated. The LTB4:LTB5 ratio shifted from 7.65 +/- 1.75 in control-fed animals to 1.03 +/- 0.18. Fish-oil-supplementation did not compromise engraftment or stem cell content. Alone, this therapy was unable to modify GvHD. PMID- 8415806 TI - Evening primrose oil and fish oil in non-insulin-dependent-diabetes. AB - The effects of two anti-thrombotic and anti-lipidemic oils, evening primrose oil and fish oil, on glucose and lipid metabolism, prostaglandin (PG) levels and body composition were studied in patients with non-insulin-dependent diabetes. Seven patients were administered 4 g evening primrose oil, 2.4 g sardine oil and 200 mg vitamin E for 4 weeks. Fasting plasma glucose, hemoglobin A1c, total cholesterol, body weight and % body fat mass were significantly decreased after the treatment, and levels of changes in these parameters were not different from 11 patients who did not receive the oils. In the treatment group, concentrations of (e) icosapentaenoic acid (EPA) increased significantly in all the lipoprotein fractions, but dihomo-gamma-linolenic acid (DGLA) increased only in the high density lipoprotein (HDL) fraction. The treatment decreased urinary 11-dehydro thromboxane B2 excretion (32.7% decrease, P < 0.05), but did not alter significantly plasma PGE1 or 6-keto-PGF1 alpha levels. The ratio of 6-keto-PGF1 alpha and PGE1 to 11-dehydro-thromboxane B2 increased significantly after the treatment. These results suggest that these oil treatments are useful in improving abnormal lipid and thromboxane (TX)A2 metabolism in diabetic patients. PMID- 8415807 TI - Role of leukotrienes and platelet activating factor in allergic bronchoconstriction and their interactions in guinea pig airway in vivo. AB - Membrane-derived lipid mediators have been considered to play a major role in pathogenesis of bronchial asthma. Recently specific antagonists and synthetase inhibitors of some chemical mediators have been developed and many studies on their anti-asthmatic effects are ongoing. But the importance of and the interactions of each mediator are still unclear. We examined the role of leukotrienes (LTs) and platelet activating factor (PAF) in immediate asthmatic response (IAR) and interactions between these lipid mediators in guinea pig airways in vivo using a specific LTs antagonist AS-35 and a specific PAF antagonist Y-24180. We confirmed the activity of each antagonist, as AS-35 and Y 24180 inhibited bronchoconstriction induced by respective agonist inhalation. AS 35 inhibition IAR but Y-24180 did not, indicating that LTs play a major role in IAR but PAF does not. AS-35 did not influence PAF-induced bronchoconstriction and Y-24180 did not inhibit LTs-induced bronchoconstriction, showing that there is no interaction between LTs and PAF. PMID- 8415808 TI - Effects of a garlic-derived principle (ajoene) on aggregation and arachidonic acid metabolism in human blood platelets. AB - When garlic cloves are chopped or crushed several dialkyl thiosulfinates are rapidly formed by the action of the enzyme alliin lyase or alliinase (EC 4.4.1.4) on S(+)-alkyl-L-cysteine sulfoxides. Allicin (diallyl thiosulfinate or allyl 2 propene thiosulfinate) is the dominant thiosulfinate released. A variety of sulfur containing compounds are formed from allicin and other thiosulfinates depending on the way in which garlic is handled. One such compound identified recently is ajoene which has been reported to possess antithrombotic properties. We present here data on the antiplatelet properties of ajoene together with its effects on the metabolism of arachidonic acid (AA) in intact platelets. Thus, ajoene was found to inhibit platelet aggregation induced by AA, adrenaline, collagen, adenosine diphosphate (ADP) and calcium ionophore A23187; the nature of the inhibition was irreversible. In washed platelets stimulated by labelled arachidonate, ajoene inhibited the formation of thromboxane A2; 12-lipoxygenase product(s) were reduced at higher ajoene concentrations. This garlic-derived substance inhibited the incorporation of labelled AA into platelet phospholipids at higher concentration. In labelled platelets, on stimulation with either calcium ionophore A23187 or collagen, reduced amounts of thromboxane and 12-HETE (12-hydroxyeicosatetraenoic acid) were produced in ajoene-treated platelets compared to control platelets. This substance had no effect on the deacylation of platelet phospholipids. The results suggest that at least one of the mechanisms by which ajoene shows antiplatelet effects could be related to altered metabolism of AA. PMID- 8415809 TI - Direct interactions between platelets and cultured rat mesangial cells. AB - Platelets seem to be involved in the pathogenesis of some kidney diseases, but the exact relationships between platelets and the changes in renal function are incompletely known. Mesangial cells (MC) were incubated with platelet supernatants (PS) and cellular surface area (CSA) and myosin light-chain phosphorylation (MLCP) were measured. CSA of PS-incubated MC (PS-MC) significantly diminished, as compared to control MC (70 +/- 6% vs. 100 +/- 5%). PS induced a significant increase in MLCP with respect to control cells (150 +/- 23% vs. 100 +/- 18%). When platelets were pretreated with indomethacin, the PS dependent contraction was abolished. Pretreatment with sulotroban (SU) or BN 52021 (BN), a thromboxane A2 (TXA2) and a platelet-activating factor (PAF) receptor blocker respectively, also completely blocked the PS effects. In other experiments, platelets were activated with thrombin (T), adding the so obtained PS to MC. Moreover, cells were also preincubated with T and then added PS. No changes in CSA were observed in either case. It may be concluded that PS contracted cultured MC, and these changes could be related to the decreased glomerular filtration rate (GFR) observed in some diseases in which platelets seem to be involved. TXA2 and PAF may be responsible for this effect. In contrast, T incubation inhibited the effect of PS, perhaps through a direct relaxing effect of T in MC. PMID- 8415810 TI - Oxy-radicals, lipid peroxides and essential fatty acids in patients with glomerular disorders. AB - It is now believed that free radicals and eicosanoids participate in the pathogenesis of immune mediated mesangial cell injury and renal tissue damage including glomerulonephritis. But relatively little information is available about the role of essential fatty acids, (EFAs), the precursors of eicosanoids, in renal diseases. We studied the levels of superoxide anion, hydrogen peroxide, lipid peroxides and the concentrations of various metabolites of EFAs in the plasma phospholipid fraction of patients with glomerular disorders. A significant increase in the production of superoxide anion and hydrogen peroxide both by unstimulated and phorbol-myristate acetate (PMA)-stimulated leukocytes, increase in the plasma lipid peroxide levels and a marked reduction in the levels of arachidonic acid (AA) and eicosapentaenoic acid, the precursors of 2 and 3 series prostaglandins (PGs) respectively, were observed in these patients. Since eicosapentaenoic acid and prostacyclin, which is derived from AA, are believed to be of benefit in the prevention of progression of renal disease, and as free radicals can cause renal dysfunction, our results suggest that free radicals and the metabolites of EFAs play a significant role in the pathogenesis of glomerular disorders. PMID- 8415811 TI - Synthesis of 12-hydroxyeicosatetraenoic acid by human endometrium and decidua. AB - In order to clarify the possible involvement of lipoxygenase metabolites of arachidonic acid (AA) in implantation, monohydroxylic acid production was examined using human endometrium and decidua in early pregnancy. As 14C-AA was mainly converted to 12-hydroxyeicosatetraenoic acid (12-HETE) as well as prostaglandins (PGs) by the microsomal fraction of the endometrium, a subsequent study was carried out on the endometrium (at various stages) and decidua in early pregnancy using tissue incubation and high performance liquid chromatography (HPLC) analysis for 12-HETE. The amounts of 12-HETE released into the culture medium by proliferative and secretory endometrium were 165.1 +/- 123.4 (ng/mg.protein/24 h, mean +/- SD) and 502.3 +/- 127.2, respectively, whereas, decidua produced a smaller amount of 12-HETE that declined as gestation progressed. This lipoxygenase product as well as PGs might have a role in implantation. PMID- 8415812 TI - Inhibition of the thermogenic and pyrogenic responses to interleukin-1 beta in the rat by dietary N-3 fatty acid supplementation. AB - The thermogenic (increase in oxygen consumption, VO2) and pyrogenic (Tc) responses to the cytokine interleukin-1 beta (IL-1 beta) were studied in rats fed a n-3 fatty acid supplemented diet (8.75% n-3 fatty acids/kg diet). 4-6 weeks after commencing the diets, the n-3 supplemented rats exhibited reduced pyrogenic (0.5 +/- 0.1 degrees C versus 1.1 +/- 0.2 degrees C in control animals) and thermogenic (9 +/- 3% versus 22 +/- 6% in control animals) responses to intraperitoneal (i.p.) injection of IL-1 beta (1 micrograms/rat). However, responses to centrally administered IL-1 beta (5ng intracerebroventricular (i.c.v.)) were similar in both groups at this time. After 8-9 weeks of supplementation, n-3 supplemented animals exhibited attenuated responses to both ip IL-1 beta (VO2 responses reduced by 68% and Tc by 0.8 degrees C) and also i.c.v. IL-1 beta (VO2 responses reduced by 56% and Tc by 0.7 degrees C). N-3 supplementation did not, however, influence the thermogenic capacity of these animals since responses to noradrenaline were similar in control and n-3 fed animals (50% increase in VO2). These findings demonstrate that n-3 supplementation modifies the pyrogenic and thermogenic responses to IL-1 beta, probably via changes in eicosanoid metabolism. Modification of central responses to IL-1 are delayed compared to the effects of peripheral administration indicating separate mechanisms of IL-1 on fever and thermogenesis in the brain and the periphery. PMID- 8415813 TI - The effect of prostaglandin E1 on renal function after cardiac surgery involving cardiopulmonary bypass. AB - This study was performed to evaluate the effect of prostaglandin E1 (PGE1) on renal function after cardiac surgery in patients undergoing cardiopulmonary bypass (CPB). Haemodynamic and renal functional response to low dose PGE1 (0.02 microgram kg-1 min-1) (group A) or saline (group B) infusion via peripheral vein during CPB was evaluated in 20 patients who underwent cardiac surgery. The perfusion pressure was maintained at about 60 mmHg during CPB in both groups. Urine beta 2-microglobulin (UBMG) (P < 0.01), and urine N-acetyl-beta-D glucosaminidase (NAG) (P < 0.05) demonstrated significantly high values after CPB in both groups compared with the presurgical value. Free water clearance (CH2O) decreased significantly at the first postoperative day compared with the pre surgical value in both groups (P < 0.01). Statistical analysis of NAG, UBMG and CH2O demonstrated significant differences between both groups, in CH2O values at the third (P < 0.05) and fifth days (P < 0.05) after surgery, in NAG values at the fifty (P < 0.01) and seventh days (P < 0.01), and in UBMG values at the first (P < 0.05) and third (P < 0.05) postoperative day, respectively. Cardiac output (co) did not change in either group throughout this study. Pulmonary capillary wedge pressure (PCWP) in group A decreased significantly at 30 and 60 min after CPB, but in group B did not change throughout the study. Cardiac index (CI) decreased significantly at 60 min after CPB in group A (P < 0.05) and at 30 min in group B (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415814 TI - The effect of nordihydroguaiaretic acid on leukotriene C4 and prostaglandin E2 production following different reperfusion periods in rat brain after forebrain ischemia correlated with morphological changes. AB - Leukotriene C4 (LTC4) and prostaglandin E2 (PGE2) are the 5-lipoxygenase and cyclooxygenase metabolites of arachidonic acid (AA). They constrict blood vessels and enhance vascular permeability inducing vasogenic edema that may hurt the ischemic penumbra after cerebral ischemia and reperfusion. Nordihydroguaiaretic acid (NDGA) is known as the most potent inhibitor of 5-lipoxygenase in different tissues. Furthermore, it has considerable inhibitory activity against cyclooxygenase. In this study, after developing a global ischemic model in the rat, the levels of LTC4 and PGE2 in the forebrain were measured, following different reperfusion periods after 10 min ischemia including 8 rats for each reperfused group. Sham operations were performed for each corresponding control group (n = 8). AA metabolites were then correlated with neuropathological findings. In the combined reperfused groups both metabolites increased significantly when compared with 10 min, ischemic group (P < 0.05). In the 8 min reperfused group, PGE2 and LTC4 increased significantly compared with each corresponding control group (P < 0.005). These mediators also increased to high levels compared with the 4 min reperfused group (P < 0.05, P < 0.005). PGE2 and LTC4 were reduced significantly at the 15th and 60th min of reperfusion compared with the 8 min reperfused group (P < 0.05, P < 0.005). NDGA (0.1 mg/kg) reduced both metabolites in the 8 min reperfused group significantly (P < 0.05). Brain cortex specimens were taken for light and electromicroscopical investigations. No significant differences were noted between the structural changes in the 4, 8 and 15 min of reperfusion and NDGA administered groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415815 TI - The 5-HT3 antagonist zacopride attenuates cocaine-induced increases in extracellular dopamine in rat nucleus accumbens. AB - Pretreatment with the serotonin-3 (5-HT3) antagonist racemic (+/-)-Zacopride hydrochloride (ZAC, 0.1 mg/kg, IP) has been previously found to completely abolish the locomotor activity induced by cocaine (10 mg/kg, IP). To determine if this effect was mediated by fluctuations in the extracellular levels of forebrain dopamine (DA), we examined the ability of ZAC to attenuate cocaine-induced increases in extracellular DA levels. Microdialysis samples were collected from the nucleus accumbens region (NAS) of awake, male, Sprague-Dawley rats. ZAC treatment alone (0.1 mg/kg, IP) did not alter DA levels relative to baseline. However, this dose of ZAC given 1 h prior to cocaine challenge (10 mg/kg, IP) caused a 27% reduction in the peak level of extracellular DA produced by cocaine, relative to saline-pretreated control animals. These results suggest that the ability of ZAC to attenuate cocaine-induced increases in extracellular DA levels may contribute to ZAC's ability to suppress cocaine-induced locomotor activity in the rat. However, additional neurochemical mechanisms are likely to be important in mediating the robust behavioral effects previously reported. PMID- 8415816 TI - Chronic, combined treatment with desipramine and mianserin: enhanced 5-HT1A receptor function and altered 5-HT1A/5-HT2 receptor interaction in rats. AB - A greater percentage of depressed patients respond to combined treatment with a tricyclic antidepressant and the tetracyclic antidepressant mianserin than to treatment with these drugs given alone. The functional sensitivity of the 5 hydroxytryptamine (5-HT)1A receptor, and the functional interaction between the 5 HT1A and the 5-HT2 receptors were investigated after treatment with desipramine and mianserin either alone or combined for 21-28 days. Pretreatment with desipramine and mianserin in combination induced the most intense 5-HT syndrome and the greatest fall in colonic temperature after injection of the 5-HT1A agonist 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT). The rats pretreated with desipramine alone had the largest elevation of the response temperature in the increasing temperature hot-plate test after injection of 8-OH-DPAT. After the combined pretreatment with desipramine and mianserin, no enhanced functional response in these tests was found when the 5-HT1A and the 5-HT2 receptors were stimulated simultaneously using 8-OH-DPAT and the 5-HT2 agonist, (+/-)-1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane HCl (DOI), contrasting the findings for desipramine or mianserin treatments given alone, where an increased functional response was found for the colonic temperature and the response temperature in the increasing temperature hot-plate test. In vitro receptor binding using [3H]-8 OH-DPAT as ligand revealed an increase in Kd and Bmax in the spinal cord after chronic treatment with the combination of desipramine and mianserin. In the hippocampus and the frontal cortex the changes were small.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415817 TI - Diurnal rhythms of 5-HT1A and 5-HT2 receptor binding in euthermic and torpor prone deermice, Peromyscus maniculatus. AB - Deermice display both spontaneous and induced daily torpor bouts, attaining minimum body temperatures of 15-20 degrees C. There is evidence that brain serotonin may be involved in the initiation and/or maintenance of torpor. Inhibition of serotonin [5-hydroxytryptamine (5-HT)] synthesis markedly reduces the duration and depth of torpor. Because a certain percentage of deermice will not enter torpor under any circumstances, we were able to compare 5-HT receptor subtypes in deermice that readily enter into torpor (TP) and in non-torpor prone (NTP) animals. Deermice were trapped in the wild and subjected to food rationing and low ambient temperature and then sacrificed either in a normothermic or torpid state at 11:00 p.m. or 11:00 a.m. Whole brain was assayed for 5-HT1A and 5 HT2 receptor differences using [3H]8-OH-DPAT and [3H]ketanserin, respectively. The Bmax values for 5-HT1A receptors were significantly greater in both TP and NTP animals sacrificed at 11:00 p.m. compared to animals sacrificed at 11:00 a.m. In contrast, the density of 5-HT2 receptors was significantly greater in animals sacrificed at 11:00 a.m. compared to animals sacrificed at 11:00 p.m. This is consistent with the opposing functions of these receptors in the regulation of temperature and sleep. The affinity (Kd) of each receptor was unchanged. A comparison of TP and NTP animals sacrificed at the same time of day revealed no significant differences in either Bmax or in Kd values, indicating that differences in 5-HT1A and 5-HT2 receptors may not explain the heterogeneity of deermice in their ability to enter torpor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415818 TI - A genetic comparison of behavioral actions of ethanol and nicotine in the mirrored chamber. AB - Human alcoholics are almost invariably heavy users of tobacco, perhaps because both ethanol and nicotine may have anxiolytic activity. However, studies in humans have not uniformly detected anxiolytic effects because significant individual differences in anxiolytic actions of these agents seem to exist. One factor that seems to contribute to these individual differences is tolerance to ethanol. Individuals who are more sensitive to depressant actions of alcohol seem to show anxiolytic actions more readily. Consequently, we examined the relative sensitivities of the ethanol-sensitive (to the anesthetic actions of ethanol) long-sleep (LS) and ethanol-resistant short-sleep (SS) mouse lines to diazepam, ethanol, nicotine, and ethanol-nicotine combinations in the mirrored chamber test. This test measures approach-conflict behavior. Ethanol and nicotine evoked changes in mirrored chamber activities that resembled those elicited by diazepam. These effects were seen at doses that did not markedly affect locomotor activity, thereby suggesting that these changes in behavior represent anxiolytic actions. The LS-SS mice did not differ in sensitivity to diazepam, but the SS were more uniformly responsive to the other drugs. Only the SS showed clear evidence for interactions between ethanol and nicotine. If the changes in mirrored chamber behavior elicited by ethanol, nicotine, and combinations of the two drugs occur because of anxiety reduction, it seems that the SS mouse line is more responsive to anxiolytic actions of these drugs. PMID- 8415819 TI - Forebrain circumventricular organs mediate captopril-enhanced ethanol intake in rats. AB - Chronic peripheral treatments with low doses of the angiotensin-converting enzyme inhibitor, captopril, enhance daily intakes of dilute ethanol solutions in rats as they do the intakes of water and saline solutions. Placing captopril into the drinking water or infusing it SC increases daily intake of 6% (v/v) ethanol from 30-100% over 4-12 days of treatment. The present study examined the effects of electrolytic lesions either of the subfornical organ (SFO) or of the organum vasculosum laminae terminalis (OVLT), on captopril-enhanced ethanol intake. Captopril was infused in minipumps at 5 mg/day for 14 days. The intake of 6% (v/v) ethanol was abolished by SFO lesions and was temporarily reduced by OVLT lesions. The SFO, in particular, is essential for the expression of enhanced ethanol intake during low-dose peripheral captopril administration. Local angiotensin II synthesis and receptor activation at the SFO appear to be the mechanism of the enhanced ethanol drinking during captopril. PMID- 8415820 TI - Pica in rats is analogous to emesis: an animal model in emesis research. AB - Mitchell et al. (1976, 1977) suggested that pica, eating of nonnutritive substances such as kaolin, is an illness-response behavior in rats. In the present study, we first confirmed their suggestion and then examined the effects of antiemetics on emetic-induced pica in rats. Intraperitoneal injection of apomorphine induced dose-dependent kaolin consumption. Pretreatment with domperidone inhibited apomorphine-induced kaolin intake. Oral administration of copper sulfate and intraperitoneal injection of cisplatin also induced dose dependent kaolin consumption. Pretreatment with ondansetron inhibited cisplatin induced kaolin intake. These findings suggest that pica in rats was induced through 1) dopamine D2 receptors in the chemoreceptor trigger zone, and 2) the stomach, partly via 5-HT3 receptors in the visceral afferents in the stomach wall. The present findings support the conclusion that pica in rats is analogous to vomiting in other species and suggest that pica in rats is mediated by the same mechanisms as vomiting in humans. Accordingly, we extended the utility of the animal model to pharmacological research of emesis with pica as an analogue to emesis. PMID- 8415821 TI - Effects of water immersion stress on convulsions induced by pentylenetetrazol. AB - Increasing durations of water immersion stress were associated with increased serum corticosterone levels, decreased severity of pentylenetetrazol-induced convulsions, and increased latencies to convulse in rats. The results were interpreted in terms of the effects of stress and pentylenetetrazol's actions on GABAergic mechanisms. PMID- 8415822 TI - Stressor controllability, social interaction, and benzodiazepine systems. AB - Effects of benzodiazepine receptor-active compounds on inescapable shock-produced changes in social interaction were studied in the rat. Inescapably shocked animals exhibited less social interaction in a novel situation than did escapably shocked or unshocked rats 24 h after shock. Administration of the selective benzodiazepine receptor antagonist flumazenil at the time of shock prevented the decrease in social interaction. Social interaction was unaffected by the same treatment at the time of measurement. Reduction in social interaction induced by inescapable stress endured for 48-72 h following stressor exposure but was absent 168 h after stress. It was subject to antagonist blockade at all measured time points. Stress-induced decreases in social interaction were also blocked by the benzodiazepine chlordiazepoxide given at the time of shock treatment. The receptor antagonist did not reverse this blockade. An inverse agonist, the beta carboline FG 7142, administered in place of inescapable shock, produced an identical pattern of social interaction in a dose-dependent manner. The inverse agonist effect was also reversed by the antagonist. The results from antagonist, agonist, and inverse agonist treatments all suggest that an endogenous benzodiazepine receptor inverse agonist is released at the time of inescapable shock and is involved in producing the changes in social interaction subsequently measured. PMID- 8415823 TI - Pargyline-induced increase in serotonin levels: correlation with inhibition of lordosis in rats. AB - The effect of intrahypothalamic infusion of the monoamine oxidase inhibitor pargyline on lordosis behavior and monamine levels in the preoptic area and hypothalamus was examined. Progesterone-facilitated lordosis was blocked by pargyline in half the treated rats. The inhibition of lordosis was correlated with increases in serotonin and dopamine levels in the ventromedial nucleus of the hypothalamus and serotonin levels in the arcuate nucleus-median eminence when compared to controls or pargyline-treated rats with high levels of lordosis responding. Changes in norepinephrine levels were not correlated with changes in behavior. The results provide further evidence for an inhibitory role of basomedial hypothalamic serotonin in the control of female sexual behavior. PMID- 8415824 TI - Respiratory patterning following cerebral ventricular administration of cocaine. AB - Intravenous (IV) cocaine in the conscious cat causes extreme tachypnea and reduction in breath-to-breath variability. In this study, we examined respiratory patterning following administration of cocaine into the cerebral ventricles. Intraventricular cocaine elicited a tachypnea that was nearly identical to that for IV cocaine. At the high dose, peak respiratory rate increased by 380%. Breath to-breath variability was dramatically reduced by cocaine, especially in the early stages of the intoxication; during these stages, the tachypnea was occasionally interrupted by prolonged inspiratory efforts. Procaine was administered as a control for the anesthetic effects of cocaine and caused an initial tachypnea that was similar to that for cocaine. For both cocaine and procaine, the mean ratios of inspiratory to expiratory durations were unaffected, indicating that the tachypnea was accomplished by approximately equal reductions in inspiratory and expiratory durations. We conclude that the tachypnea following cocaine administration results principally from central rather than peripheral mechanisms. In addition, the data suggest that anesthetic actions mediate the principal respiratory effects of cocaine. PMID- 8415825 TI - Arrest of seizure series induced by an intracortical injection of penicillin in the awake rat. AB - Experiments were performed to answer the question, whether series of generalized tonic-clonic seizures, induced in the awake rat by a local injection of Na penicillin (PCN) solution into the motor cortex, terminates at the same critical concentration Ct of PCN within the focal area independently of the concentration C0 of PCN injected. Using the PCN diffusion coefficient D = 3.52 x 10(-4) mm2/s and the tortuosity factor lambda = 1.62, the concentration Ct at the onset of the last generalized seizure was calculated. The median duration of seizure series increased from 32 to 190 min, when the dose of injected PCN was raised from 32 to 1000 IU. At the onset of the last seizure, the median concentration Ct within the artificial focus ranged from 2.1 to 4.0 IU/0.5 microliter saline in rats treated with 32 to 125 IU PCN. After induction of convulsive behaviour with C0 = 250, 500, or 1000 IU PCN/0.5 microliter saline, however, Ct was at a higher level between 6.1 and 7.4 IU PCN/0.5 microliter saline. The difference between the cumulated data from the low-dose vs. the high-dose range was significant (p < 0.01). It is concluded that during long-lasting series of generalized seizures, the brain takes advantage of its plastic properties. By forming a counteracting mechanism, it protects itself from extreme epileptiform activity. This autoprotection may be due to the activation of neuronal networks which probably needs a certain frequency of seizures to become operatively. PMID- 8415826 TI - Intrathecal coadministration of serotonin and morphine differentially modulates the tail-flick reflex of intact and spinal rats. AB - In a previous study, we found that the antinociceptive effect of IT-administered morphine on the tail-flick (TF) reflex of rats was potentiated within 1 day after spinal transection. This suggested that the analgesic effect of spinal morphine in the intact animal was tonically suppressed, presumably by the release of a transmitter(s) from descending supraspinal pathway(s), and that the potency of IT morphine was increased because these inputs were removed by spinalization. Because spinally projecting serotonin [5-hydroxytryptamine (5-HT)] fibers are known to be involved in modulating nociception at this site, the present studies examined the possibility that 5-HT might be the proposed "antiopiate" at the spinal cord. Separate groups of intact and spinal rats were pretested on the TF and then injected IT with either morphine (intact: 0.25-5.0 micrograms, spinal: 0.0312-0.5 microgram) or 5-HT (1-200 micrograms), or combinations of these two agents, in a single solution. All rats were then retested 15 min later and the difference in latency was used to compare the effect of these treatments. The results confirmed that the antinociceptive effect of IT morphine was significantly increased by spinalization, whereas the antinociceptive effect of 5 HT was essentially abolished. In intact rats, morphine-induced analgesia was potentiated by a low (10 micrograms) dose of 5-HT but not by higher doses. However, in the spinal rat morphine-induced antinociception was antagonized by the same (10 micrograms) dose. The data suggest that IT 5-HT promotes antinociception in intact rats but acts pro-nociceptively in spinal rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415827 TI - Attenuation of anticonvulsant effects of diazepam after chronic treatment with bicuculline. AB - Changes in the GABAergic system after chronic treatment with bicuculline were examined in two strains of inbred rats, Fischer 344 (F344) and Lewis (LEW). Rats received an IP injection of either bicuculline (2 mg/kg) or vehicle once a day for 12 days. After this chronic treatment, the effects of diazepam (1 mg/kg, IP) and pentobarbital (20 mg/kg, IP) on bicuculline-induced convulsions were measured. Bicuculline was acutely infused into a tail vein at 0.0415 mg/min, and the infusion was terminated when rats showed seizure. Following the chronic bicuculline treatment, the anticonvulsant effect of diazepam, but not of pentobarbital, was significantly reduced as compared to its effect following chronic vehicle treatment in both strains. Both diazepam and pentobarbital showed a significant difference in anticonvulsant effects between strains (F344 > LEW). The hypnotic effects of muscimol, barbital, pentobarbital, and ethanol following chronic bicuculline treatment were examined. There was no significant difference in sleep time induced by these drugs between bicuculline- and vehicle-treated rats. These results suggest that the attenuation of diazepam's anticonvulsant effect after chronic bicuculline treatment may result from functional changes in benzodiazepine receptors and that the anticonvulsant effects of diazepam and pentobarbital may be influenced by genetic factors. Moreover, the hypnotic effects of several drugs tested are apparently not affected by chronic bicuculline treatment. PMID- 8415828 TI - Models of simple partial and absence seizures in freely moving rats: action of ketamine. AB - The action of ketamine was studied in two models of seizures: a) bilateral neocortical discharges produced by topical application of pentylenetetrazol (model of simple partial seizures); and b) rhythmic spike-and-wave activity induced by systemic administration of pentylenetetrazol (model of absence seizures). Ketamine exerted biphasic effects. In the first model, the dose of 20 mg/kg ketamine significantly suppressed the ictal neocortical discharges (i.e., continuous spiking or ictal activity) accompanied by clonic motor seizures. However, at the dose of 40 mg/kg ketamine significantly accentuated the onset and increased the number of individual discharges (interictal spikes) in bilateral neocortical foci. In the model of rhythmic spike-and-wave activity, the lower dose of ketamine (20 mg/kg) decreased the number of rhythmic spike-and-wave episodes when compared to the higher dose (40 mg/kg) of ketamine, which increased the number of episodes. However, neither result differed significantly from control values. The present results suggest a dose-dependent action of ketamine: Lower doses (10 and 20 mg/kg in the rat) are able to suppress seizure activity, whereas a higher dose (40 mg/kg) potentiates the seizures. Moreover, the action of ketamine may be dependent upon the seizure model used. The study presents a new model of acute epileptic focus in freely moving rats. PMID- 8415829 TI - Strain and housing affect cocaine self-selection and open-field locomotor activity in mice. AB - We recently conducted an experiment to investigate the possible cooperation between genetic makeup and differential housing on cocaine self-administration in male and female C57BL/6J and DBA/2J mice. Cocaine self-selection was measured in a two-choice test with one choice being cocaine-HCl solution of 40 mg% in tap water and the other choice being plain tap water. Housing conditions began at weaning (21-23 days of age) and consisted of group housed (GH) with 2-3 mice per cage, and isolated housed (IH) with 1 mouse per cage. The results of this study revealed overall strain, sex and housing differences, with C57BL/6Js consuming more cocaine solution than DBA/2J subjects, females consuming more cocaine solution than males, and group housed consuming more than isolate housed subjects. In a second study, the effect of differential housing on open-field locomotor activity was investigated. Testing was conducted on two consecutive days, with subjects receiving an IP injection of saline on day 1, and 15 mg/kg cocaine HCl on day 2. Four behaviors were recorded, including: total distance, nosepokes, stereotypy, and margin time. Overall, the results revealed significant strain differences for stereotypy and nosepokes, and males were found to be more activated by cocaine than females. Additionally, DBA males tended to be differentially affected by housing condition, with IH showing suppressed locomotor activity as compared to GH subjects. Last, significant strain by housing interactions occurred in nosepokes and stereotypy time. PMID- 8415830 TI - Adenosinergic modulation of the EEG and locomotor effects of the A2 agonist, CGS 21680. AB - The present study in rats was designed to identify the respective roles of A1 and A2 adenosine receptor activation in the anticonvulsant and behavioral actions of adenosine. Intracaudate injections of the highly selective A2 agonist, CGS 21680, did not affect caudate seizures. However, seizure threshold was increased in the presence of CGS 21680 after blockade of the A1 receptor with CPX, or following activation of the A1 receptor with R-PIA or NECA. Additionally, CGS 21680 led to a dose-related inhibition of locomotor activity when injected into the caudate. These results implicate the involvement of the A2 adenosine receptor in the locomotor depressant actions of adenosine and also suggest possible A2 anticonvulsant effects may depend upon the activation of the A1 receptor. PMID- 8415831 TI - A subpopulation of dopamine D1 receptors mediate repetitive jaw movements in rats. AB - Repetitive jaw movements (RJM) in rats, a potentially useful animal model of tardive dyskinesia, appears to be mediated by the dopamine D1 receptor as evidenced in part by their induction and inhibition with D1 agonists and D1 antagonists, respectively. Selective destruction of 60-90% of D1 receptors by EEDQ, measured in several CNS dopaminergically innervated areas, preceded by protection of D2, 5-HT2, alpha 1 and alpha 2 receptors, however, failed to reduce D1 agonist-augmentable RJM. Further, the affinity of dopamine toward displacement of 3H-SCH-23390 binding from striatal D1 receptors was significantly decreased by administered EEDQ, a counter-intuitive result in relation to D1 responsitivity and RJM. Thus, at present it is suggested that an EEDQ-resistant D1 receptor subpopulation may exist. PMID- 8415832 TI - Scopolamine reversal of nicotine enhanced delayed matching-to-sample performance in monkeys. AB - The basis for the memory enhancing action of nicotine was evaluated in five adult monkeys (Macaca fascicularis) well trained in the performance of a delayed matching-to-sample (DMTS) paradigm. Nicotine (1.25-20 micrograms/kg, IM) produced a dose-dependent improvement in performance of the task. The optimal dose of nicotine for each monkey also improved performance when the animals were tested 24 h later in the no-drug situation. In the same animals, low doses of scopolamine produced a dose-dependent decrement in DMTS performance. A subthreshold dose (defined by DMTS performance decrement) of scopolamine was administered 20 min prior to the optimal dose of nicotine. Scopolamine pretreatment completely blocked the enhanced performance observed earlier with nicotine. The results of this study are consistent with the hypothesis that the enhanced cognitive performance associated with nicotine is due to central acetylcholine release and subsequent muscarinic receptor stimulation. PMID- 8415833 TI - Novel environment and cat odor change GABA and 5-HT release and uptake in the rat. AB - In hippocampal and cortical slices taken from rats moved in their home cages to a novel environment for 5 min, there were decreases in basal and K(+)-evoked [14C]GABA release and an increased [14C]GABA uptake compared with slices taken from rats remaining undisturbed in the animal house. The changes in 5 hydroxytryptamine (5-HT) release and uptake in response to a novel environment were markedly time dependent. In rats killed immediately after the 5-min exposure, there was decreased hippocampal [3H]5-HT uptake and higher hippocampal basal release, whereas in rats killed 30 min later there was increased [3H]5-HT uptake and lower basal release in both hippocampal and cortical slices. Rats exposed to cat odor in the novel environment showed increased release and decreased uptake of GABA in both brain areas compared with the group exposed to a neutral odor in the same novel environment, and these differences between the two odor groups were found both immediately and 30 min after the odor exposure. In contrast, only one measure of 5-HT function differed between the neutral and cat odor groups, with the latter showing increased cortical [3H]5-HT uptake 30 min after odor exposure. PMID- 8415834 TI - Thermoregulatory responses following injection of 5-hydroxytryptamine into the septohippocampal complex in rats. AB - The present study investigated the change in thermoregulatory responses following microinjection of 5-hydroxytryptamine (5-HT) into the lateral septum and the hippocampus of unanesthetized, unrestrained rats. Intraseptal injection of 5-HT (5 to 20 micrograms) caused a dose-related fall in core temperature (Tb), which was associated with a decrease in heat production (HP). As the decrease in HP can not completely account for the magnitude of the decrease in Tb, increase in heat loss may also be involved in the 5-HT-induced hypothermia. In contrast to observed changes following intraseptal injection, no significant change in either Tb or HP was observed after microinjection of the same doses of 5-HT into the hippocampal areas, indicating that the hypothermic response to intraseptal injection of 5-HT is site specific. Further, the hypothermic response to intraseptal injection of 5-HT was only attenuated by systemic pretreatment with cyproheptadine, but not by naloxone or scopolamine, indicating that the hypothermic response is mediated by 5-HT receptor, but not by endogenous opioid and cholinergic systems. PMID- 8415835 TI - Effect of naloxone and antidepressants on hyperphagia produced by peptide YY. AB - Central injection of peptide YY (PYY) elicits a powerful feeding response with a short latency in satiated rats. Because of this effect, PYY has been implicated as a neurochemical signal in bulimia nervosa. Serotonin agonists and opioid antagonists induce anorectic effects upon feeding behavior in humans and animals. Therefore, to investigate a possible interaction between PYY-induced eating and these anorexigenic agents rats were given injections of either naloxone (100 micrograms/3 microliters, ICV, and 10 mg/kg, SC), fluoxetine (3-30 micrograms/3 microliters and 5-10 mg/kg, IP), or clomipramine (3-30 micrograms/3 microliters and 5-10 mg/kg, IP) prior to fourth ventricular injections of PYY (15 micrograms/18 microliters). Central and peripheral naloxone and IP but not central injections of fluoxetine blocked PYY-induced intake. Clomipramine had no effect. This suggests that PYY-stimulated feeding may require the action of endogenous opioids and may be inhibited by serotonergic function. PMID- 8415836 TI - In utero exposure to fluoxetine HCl increases hematoma frequency at birth. AB - The present study was undertaken to determine if fluoxetine HCl (Prozac, Dista Products Ltd., Liverpool, UK) might cause adverse vascular effects, such as hematomas, in rats exposed in utero. Gravid Sprague-Dawley rats were administered 5.62 mg/kg fluoxetine HCl by oral gavage beginning on day 7 of gestation and ending the day of birth. A control group received distilled water by oral gavage during gestation. At birth, offspring of both groups were assessed for visible adverse vascular effects. Fluoxetine HCl-exposed offspring showed a statistically higher frequency of skin hematomas when compared to water controls. This result is consistent with known adverse effects of fluoxetine and lends support to a recently published report that attempted to link fluoxetine HCl use to bleeding episodes in eight patients being treated for obsessive-compulsive disorder. The results of this study suggest caution in the prolonged use of this medication during pregnancy and in patients with predisposing conditions that may increase the chances of bleeding. PMID- 8415837 TI - Preliminary findings with the indirect 5-HT agonist dexfenfluramine on heroin discrimination and self-administration in rats. AB - In the present study, the effect of the 5-HT releaser/reuptake inhibitor dexfenfluramine in heroin self-administration and morphine (3 mg/kg) drug discrimination paradigms was examined. Dexfenfluramine (1 mg/kg) reduced heroin self-administration (heroin dose 0.03 mg/kg/infusion; FR5 schedule; 1-h session/day). This effect was antagonised by the 5-HT1/2 receptor antagonist metergoline (1 mg/kg). In the drug discrimination model, dexfenfluramine (0.5-2.5 mg/kg) produced no significant generalisation to a morphine cue, and also failed to modify the generalization curve to heroin. Dexfenfluramine (1 mg/kg) produced a slight decrement in response rate in the drug discrimination model and this effect was potentiated by heroin. The mechanism(s) by which dexfenfluramine reduces heroin self-administration remain to be determined, however substitution for heroin would seem unlikely. Furthermore, this effect of dexfenfluramine is probably mediated by either 5-HT1 or 5-HT2 receptors. PMID- 8415838 TI - alpha-Difluoromethylornithine does not antagonize the behavioral effects of putrescine. AB - alpha-Difluoromethylornithine (DFMO), a specific inhibitor of putrescine synthesis, is widely used in studies of polyamine function as well as clinically. We studied the effect of DFMO on the tendency to explore in the Greek cross maze that provides the rat with the choice to enter either white or black compartments. After a single injection of 400 mg/kg DFMO, the entries into white compartments were significantly decreased. A similar decrease had been observed previously with 200 mg/kg putrescine. Simultaneous administration of DFMO (400 mg/kg) and putrescine (200 mg/kg) resulted in decreased entries into both white and black compartments. When 400 mg/kg DFMO plus 400 mg/kg putrescine were injected, the entries into both compartments were further decreased and the time spent in white compartments was also decreased. This pattern mimicked that found with anxiogenic drugs. Injection of DFMO (400 mg/kg) produced no change in either putrescine, spermidine, or spermine concentration measured in brain cortex. Putrescine (200 mg/kg) plus DFMO produced the same transient increase in cortical putrescine as putrescine alone. We conclude that DFMO is mildly anxiogenic and that this activity is independent of its inhibition of putrescine synthesis. PMID- 8415839 TI - The functional relevance of the lateral parabrachial nucleus in lithium chloride induced aversion learning. AB - Lesions to the lateral parabrachial nucleus (PBN), one of the subnuclei that make up the pontine parabrachial complex, impairs the acquisition of taste aversion learning (TAL) with LiCl as the toxic stimulus. In this experiment, PBNl-lesioned and control rats were trained to learn a delayed task with a 15-min interval between presentation of the gustatory and the aversive stimulus. The impairment in learning observed after lesions of the PBNl is discussed in terms of disruption of the transmission of toxic stimuli (LiCl) processed by the humoral pathway and the area postrema (AP). PMID- 8415840 TI - Centrally administered mu- and delta-opioid agonists increase operant responding for saccharin. AB - In previous reports, ICV administration of selective mu- or delta-opioid receptor agonists was found to stimulate the intake of saccharin and salt solutions in nondeprived rats. In the present study, we measured the effects of selective mu-, delta-, and kappa-agonists on operant responding for saccharin. The selective mu agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) and the selective delta agonist [D-Thr2]-leucine enkephalin-Thr (DTLET) increased responding, whereas the kappa-agonist dynorphin A analog kappa ligand (DAKLI) had no significant effect. These results agree with previous studies on saccharin and salt intake and are consistent with the possibility that the effects of opioids on the intake of these fluids are mediated via enhancement of activity in brain reward pathways. PMID- 8415841 TI - Effect of D-cycloserine on rapid tolerance to ethanol. AB - We recently reported that the noncompetitive NMDA antagonists, (+)MK-801 and ketamine, block the development of rapid tolerance to ethanol. In the present article, we show that D-cycloserine (CS), an agonist at the glycine site of the NMDA receptor that enhances learning and memory, also enhances the development of rapid tolerance to ethanol. Rats were pretreated on day 1 with saline or CS, followed 30 min later by ethanol (2.3 g/kg, IP) or saline. At the end of motor impairment testing on the tilt-plane apparatus, a second injection of CS (3 mg/kg, IP, each time) or saline was given, followed 30 min later by ethanol or saline. Ethanol pretreatment alone (at this dose) did not result in rapid tolerance to ethanol on day 2. However, the group pretreated with CS and ethanol on day 1 showed significant tolerance on day 2 compared to other groups. Pretreatment with CS on day 1 did not affect the motor impairment response to the first exposure to ethanol whether this was on day 1 or day 2. In another experiment, administration of (+)MK-801 (0.25 mg/kg, IP) prior to CS abolished the rapid tolerance enhancement by CS. These findings are further evidence that the NMDA system, which requires activation by the glycine receptor, plays a major role in the development of at least some forms of ethanol tolerance. PMID- 8415842 TI - Effects of caffeine and PD 116,600 on the differential-reinforcement-of-low rate 72-S (DRL 72-S) schedule of reinforcement. AB - Caffeine and PD 116,600 were found to decrease the reinforcement rate and increase the response rate in rats performing under a differential-reinforcement of-low rate 72-s (DRL 72-s) schedule of reinforcement. In contrast, antidepressant drugs previously have been found to increase the reinforcement and decrease the response rate. Caffeine has been found to test similar to antidepressant drugs on at least one other behavioral screen, but caffeine does not possess clinical antidepressant properties. These results provide further support for the DRL 72-s schedule as a behavioral screen for antidepressant drugs. PMID- 8415843 TI - Glucocorticoids antagonize the sedative action of ethanol in mice. AB - The effect of corticosterone on sleep time in mice following a hypnotic dose of ethanol (3 g/kg) was determined. An acute dose of the steroid (10 mg/kg) administered 15 min prior to ethanol injection significantly shortened the sleep time (by 55%). Brain levels of ethanol were not affected by the steroid treatment. The effect was specific to glucocorticoids because steroids without glucocorticoid activity including testosterone and 17 beta-estradiol were ineffective. These results indicate that glucocorticoids have an antagonistic effect to the acute action of ethanol in the brain. The rapid onset of the corticosterone action in antagonizing ethanol-induced sedation suggests that the action is mediated by a membrane mechanism rather than the classical steroid mechanism involving an intracellular receptor and gene expression. PMID- 8415844 TI - Combined serotonergic-cholinergic lesions do not disrupt memory in rats. AB - Rats were trained on a delayed nonmatching to position task, divided into four groups and given the following lesions: (a) SHAM (vehicle injection into nucleus basalis magnocellularis (NBM) and raphe nuclei (RN), (b) RN (5,7 dihydroxytryptamine lesions of raphe, vehicle into NBM), (c) NBM (quisqualic acid lesion of NBM, vehicle into RN), and (d) COMB (lesions of both RN and NBM). RN lesions had no effect on performance measures including accuracy (percent correct), errors of omission, bias, latencies, and magazine response rate. NBM lesions produced delay-independent (nonmnemonic) disruptions, but performance improved over the 20 days' test. The effects of COMB lesions were no worse than NBM lesions alone. The results suggest that (a) the serotonergic system is not essential for performance in this task, (b) NBM lesions transiently impair nonmnemonic aspects of performance, and (c) serotonergic-cholinergic interactions may not be essential for some cognitive processes. PMID- 8415845 TI - [Agonists at the histamine H1 receptor: structure and pharmacology]. PMID- 8415846 TI - [Inhibitors of phospholipase A2]. PMID- 8415847 TI - Synthesis and positive inotropic activity of several 5-aminopyrido[2,3 d]pyrimidines. Part 5: Compounds with positive inotropic activity. AB - Starting from 6-amino-1,3-dimethyluracil two approaches were developed for the preparation of 5-amino-pyrido[2,3-d]pyrimidine derivatives as potential cardiotonic agents. 1. Gould-Jacobs reaction followed by chlorination of the intermediate 5-hydroxypyrido-[2,3-d]pyrimidine using DMF/POCl3. 2. Cyclization of C-acetylated as well as C-cyano acetylated 6-amino-1,3-dimethyluracil by an application of the Vilsmeier reaction yielding 5-chloropyrido[2,3-d]pyrimidines. Subsequent nucleophilic substitution reactions formed the target compounds which were examined for positive inotropic activity on isolated left atria and papillary muscles from guinea-pig hearts. Structure-activity relationships indicated that the effect depended on the 4-aminopyridine-3-carboxylic acid derivative structure. PMID- 8415848 TI - [Synthesis of new pyrido-(3',2':4,5)-thieno-(3,2-d)-1,2,3-triazine derivatives as antianaphylactics]. AB - Some new pyrido[3',2':4,5]thieno[3,2-d]1,2,3-triazine-4(3H)-ones (C) were synthesized from 2-thioxo-1,2-dihydro-3-carbonitriles (A) via the 3-amino thieno[2,3-b]pyridine-2-carboxamides (B). Substances of structure A were converted to 3-amino-thieno[2,3-b]pyridine-2-carbonitriles (G) which yielded the desired 4-amino substituted compounds I via the tricyclic 4-chloro pyrido[3',2':4:5]thieno[3,2-d]1,2,3-triazines (H) by the reaction with N nucleophiles. Some of the investigated compounds showed a respectable antianaphylactic activity. PMID- 8415849 TI - [Polycyclic aromatic alkaloids. 9. Synthesis and antifungal activity of diazafluoroanthene alkaloids]. AB - The total syntheses of the alkaloids eupolauridine-N-oxide (2) and eupolauridine di-N-oxide (3) are described. The compounds 2 and 3 possess lower anticandidal activity than eupolauridine (1). PMID- 8415850 TI - [The endocrinologic profile of metabolites of the progestin dienogest]. AB - Two identified metabolites of the orally active progestin dienogest, the compounds STS 749 (17 alpha-cyanomethyl-11 beta,17 beta-dihydroxy-estra-4,9-dien 3-one) and STS 825 (17 alpha-cyanomethyl-estra-1,3,5(10),9(11)-tetraene-3,17 beta diol), furthermore 4 microbially formed metabolites and 10 chemically prepared analoga of dienogest were characterized by endocrinological tests. The compounds were investigated for progesterone-receptor binding, progestational and antiprogestational, estrogenic and antigonadotropic activities, furthermore for inhibition of fertility. In no case an increased progestational activity of metabolites or analoga was found, compared to the parent substance. Therefore, it should be excluded, that dienogest acts as a prodrug. Additionally, the dosages necessary to produce the progestational effects are quite similar using oral or subcutaneous application. Therefore, a first-pass effect can be neglected. Regarding the low endocrine side effects of dienogest, the antiprogestational activity may be caused, at least in part, by metabolites. PMID- 8415851 TI - Determination of N-butyl-scopolamine bromide in human plasma and urine by a HPLC method. Evidence of negligible absorption after oral administration. PMID- 8415852 TI - Preparation, characterization and anti-inflammatory effect of defibrotide liposomes. PMID- 8415853 TI - [Cytotoxic activity of new cyclic lactones in the "yeast test"]. PMID- 8415854 TI - [Absorption and excretion of apigenin, apigenin-7-glycoside and herniarin after oral administration of extracts of Matricaria recutita (L.) (syn. Chamomilla recutita (L.) Rauschert)]. PMID- 8415855 TI - Interaction of sanguinarine with human serum albumin. PMID- 8415856 TI - [Synthesis of 11-aryl-7,8,9,10-tetrahydro-1,2,3-triazino-(4',5':4,5)-thienol- (2,3-b)-quinolines with antianaphylactic action]. AB - Title compounds of structure B with an oxo function in position 4 of the triazine ring were synthesized by reaction of the aminocarboxamides A with sodium nitrite in acetic acid. Aminocarbonitriles of structure H yielded with sodium nitrite in acetic acid and hydrochloric acid the 4-chloro derivatives I. These compounds gave with N-nucleophiles, methoxide or thiourea the substances J. Tetracyclic compounds with a hydroxyethyl or a piperidinoethyl residue in position 3 in the triazine ring (E, G) were also prepared. Some of the investigated compounds showed an antianaphylactic activity. PMID- 8415857 TI - [Synthesis and anti-inflammatory activity of some indazole derivatives. 36. Azoles]. AB - The synthesis of water soluble hydrochlorides of indazole derivatives 1b, 8 and 9 is described. By treating of 2,5-dinitroindazole with thiomorpholine 3 thiomorpholino-5-nitroindazole (10) and 3,5-dinitroindazole (11) in the form of the molecular compound 11a are obtained. The known indazole derivatives 1 and 7 as well as the newly synthesized hydrochlorides of 1b, 8 and 9 are, except of 8.HCl, less toxic than benzydamine hydrochloric (BZD). The same compounds show with some excepts a comparable or greater antiinflammatory effect than BZD in the carrageenin induced oedema test. PMID- 8415858 TI - Erythromycin-antacid interaction. AB - The in vitro release of erythromycin stearate in the presence of aluminium hydroxide, aluminium trisilicate, magnesium oxide, magnesium trisilicate, sodium hydrogen carbonate and a combination of aluminium hydroxide gel, magnesium hydroxide and dimethyl polysiloxane has been studied by the USP XX dissolution method. The dissolution of erythromycin was found to be markedly retarded in the presence of all the antacids studied except sodium hydrogen carbonate. An attempt was made to elucidate the mechanism of this effect. PMID- 8415859 TI - Evaluation of sterically stabilized liposomes as a vehicle for targeting technetium-99m labelled radiopharmaceuticals. AB - Sterically stabilized neutral liposomes (multilamellar vesicles) were prepared by sonicating phosphatidylcholine and cholesterol (molar ratio 4:1) film in phosphate buffered saline (50 mM, pH 7.4) containing 4% Tween 20. Tc-99m-GHA was incorporated in these liposomes by treating 0.5 ml of the suspension with lyophilized GHA kit (5 mg GHA and 250 micrograms SnCl2 x 2 H2O) followed by addition of 1 ml 99mTcO4- (1-3 mCi). The labelling yield was 60-70%. Tween 20 has provided significant stability of the radiolabel as compared to that without its addition, when radiolabelled liposomes were incubated in serum up to 24 h. With respect to Tc-99m-GHA alone, radiolabelled liposomes exhibited 4- to 6-fold greater radioactivity in the blood of rabbits (15 min-24 h). Comparison of biodistribution data of radiolabelled liposomes and Tc-99m-GHA in mice demonstrated a 10- to 12-fold greater hepatic accumulation of radiolabelled liposomes with respect to that of Tc-99m-GHA throughout the period of study (15 min-24 h), though their concentration in the kidneys was comparable. PMID- 8415860 TI - [Gas chromatographic determination of gamma-hydroxybutyric acid in human plasma]. PMID- 8415861 TI - Non-interaction of ranitidine with hydroxylation of diclofenac in rat and human liver microsomes. PMID- 8415862 TI - Excretion of 7-hydroxycoumarin conjugates in human urine. PMID- 8415863 TI - Inhibition of mobility of Tetrahymena pyriformis by certain new local anaesthetics: an alternative in vitro approach to drug screening. PMID- 8415864 TI - Participation of intracellular calcium stores in serotonin-induced contractions in rat aorta. AB - Serotonin 1 mumol/l induces a contractile response in the isolated rat aorta in both the presence or absence of extracellular Ca. The present study analyzes the influence of temperature and caffeine on subsequent serotonin-induced contractions. In Ca-free medium, the contraction elicited by serotonin was higher at 25 degrees C than at 37 degrees C. In addition, the existence of two independent intracellular Ca pools releasable by serotonin, one of them also sensitive to caffeine, is postulated. The results also showed that addition of serotonin decreases the contractile response to this agonist in Ca-free medium. PMID- 8415865 TI - Effects of phorbol 12,13-dibutyrate and H-7 in extravascular smooth muscle contraction. AB - The effect of the activator, phorbol 12,13-dibutyrate (PDB), and the inhibitor, H 7, of protein kinase C (PKC) has been assayed in rat uterus. PDB increases the amplitude of spontaneous contractions of rat uterus and this effect does not occur in the presence of H-7 or nifedipine. PDB did not modify the KCl-induced tonic contraction but H-7 relaxed it, in a concentration-dependent way. PDB inhibited the contraction induced by oxytocin in rat uterus incubated in Ca-free solution and relaxed the tonic contraction induced by oxytocin in this medium. The relaxing effect of PDB on oxytocin-induced contraction was not modified by H 7. Thus H-7 relaxed, in a concentration-dependent way, the tonic contractions induced by oxytocin and vanadate in the rat uterus incubated in Ca-free medium. Our results suggest a dual effect of PDB related to calcium, and a direct and PKC independent inhibitory effect of H-7. PMID- 8415866 TI - Effects of bupivacaine on contraction and membrane potential in isolated canine papillary muscles. AB - The effects of bupivacaine on myocardial contraction and membrane potential were examined in isolated canine right ventricular papillary muscles. Bupivacaine (10( 6) to 10(-4) mol/l) produced a dose-dependent and reversible decrease in the electrically induced contractile response of canine right ventricular papillary muscles. The inhibitory effects of bupivacaine on contraction were more pronounced at high stimulation frequencies (2 and 3 Hz) than at low frequency (< 1 Hz). The resting membrane potential was not affected by bupivacaine. The maximal upstroke velocity of the action potential was reduced by 10(-6) mol/l bupivacaine (74 +/- 28% of control), and these effects were also dose dependent. At 10(-4) mol/l, bupivacaine blocked fast action potentials in normal Tyrode's solution. Furthermore, bupivacaine (10(-4) and 10(-3) mol/l) decreased both slow action potential duration and associated contractions in high-K+ (26 mmol/l) Tyrode's solution in the presence of isoproterenol. Our results suggest that low concentrations of bupivacaine depress contraction mainly due to an Na+ channel block, whereas at higher concentration, this local anesthetic may block Ca2+ channels. PMID- 8415867 TI - G619, a dual thromboxane synthase inhibitor and thromboxane A2 receptor antagonist, reduces myocardial damage and polymorphonuclear leukocyte accumulation following coronary artery occlusion and reperfusion in rats. AB - We investigated the effect of G 619, a dual thromboxane synthase inhibitor and thromboxane A2 (TxA2) receptor antagonist, in pentobarbital-anaesthetized rats subjected to left main coronary artery ligation (1 h) followed by reperfusion (1 h; MI/R). Sham-operated rats were used as controls (sham MI/R). Survival rate, myocardial necrosis, myocardial myeloperoxidase (MPO) activity (investigated as an index of leukocyte adhesion and accumulation) and serum creatine phosphokinase (CPK) activity were studied. MI/R injury significantly reduced survival rate (45%), caused a marked myocardial necrosis, increased serum CPK activity (sham MI/R = 35 +/- 12 U/ml; MI/R = 205 +/- 13 U/ml) and produced an increase in myocardial MPO activity in the area at risk and in the necrotic area (6.3 +/- 0.5 and 6.6 +/- 0.9 U x 10(-3)/g tissue, respectively). The administration of G 619 significantly increased survival rate, lowered the area of necrosis, blunted the increase in serum CPK activity and reduced the increase in MPO activity in both the area at risk and the necrotic area. These data are consistent with an involvement of TxA2 in MI/R injury and suggest that G 619 may represent a novel therapeutic approach to the treatment of acute myocardial infarction. PMID- 8415868 TI - Antihypertensive activity of ABBOTT-81282, a nonpeptide angiotensin II antagonist, in the renal hypertensive rat. AB - ABBOTT-81282, 4-(N-butyl-N-[(2-'-[1H-tetrazol-5yl]biphenyl-4-yl)methyl]ami no) pyrimidine-5-carboxylic acid is a novel nonpeptide angiotensin II (AII) antagonist. In vivo studies were performed to evaluate ABBOTT-81282 for its antihypertensive effect, pharmacological mechanism(s) of action, and cardiovascular safety. In the conscious renal artery-ligated (RAL) hypertensive rat, a model of high renin hypertension, ABBOTT-81282 (1-10 mg/kg p.o. and 0.1 1.0 mg/kg i.v.) lowered mean arterial pressure (MAP) in a dose-dependent manner with the ED30 values of 2.2 mg/kg for p.o. administration and 0.08 mg/kg for i.v. administration. At 10 mg/kg p.o., ABBOTT-81282 lowered blood pressure in the RAL rat (delta MAP 66 +/- 9 mm Hg from control MAP 167 +/- 7 mm Hg, n = 6) to a normotensive level (MAP, 115 +/- 5 mm Hg) for greater than 24 h and did not change heart rate. The i.v. administration of 1 mg/kg of ABBOTT-81282 also produced a sustained, long-lasting decrease (delta MAP 27-52 mm Hg) in blood pressure that was significantly different from the vehicle group at 8 h postdosing (143 +/- 3 mm Hg, n = 4 for ABBOTT-81282 vs. 181 +/- 3 mm Hg, n = 6 for vehicle group, p < 0.01). When blood pressure in the renal hypertensive rat was maximally lowered (delta MAP 72 +/- 9 mm Hg, n = 4) following the 1 mg/kg i.v. dose (cumulative) of ABBOTT-81282, additional administration of captopril (3 mg/kg i.v.) produced no further decline in blood pressure. In the conscious normotensive rat, 10 mg/kg p.o. of ABBOTT-81282 had no effect on basal MAP (119 +/- 3 vs. 115 +/- 4 mm Hg, pre- vs. 3.5 h postdosing, n = 4) and heart rate (364 +/- 18 vs. 363 +/- 14 beats/min, pre- vs. 3.5 h postdosing, n = 4) but inhibited the AII (0.1 micrograms/kg i.v.)-induced increase in MAP by 64-70%, while the MAP responses to norepinephrine (0.3 micrograms/kg i.v.), vasopressin (0.03 IU/kg i.v.) and bradykinin (3 micrograms/kg i.v.) remained intact. ABBOTT-81282 was also administered to conscious normotensive rats (n = 4) instrumented with ECG telemetry transmitters. At an i.v. dose of 10 mg/kg, which is 125 times greater than the i.v. ED30, ABBOTT-81282 caused a minimal decrease (< 14%) in MAP and had no effect on ECG waveforms. These data demonstrate that ABBOTT-81282 is a safe and efficacious antihypertensive agent with selective AII antagonism.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8415869 TI - Attenuation of ischemic acute renal failure by phosphoramidon in rats. AB - The protective effects of phosphoramidon, a dual inhibitor of endothelin converting enzyme and neutral endopeptidase (E.C. 24.11), on renal function in ischemic acute renal failure were investigated in anesthetized rats. Intravenous infusion of phosphoramidon (0.03 and 0.1 mg/kg per min) significantly suppressed tubular sodium wasting (measured by fractional excretion of sodium) and proteinuria in the postischemic kidney without modifying functional parameters in the contralateral normal kidney. Phosphoramidon (0.1 mg/kg/min) was associated with increased glomerular filtration in the ischemic kidney. In comparison, SCH 42354, a highly selective inhibitor of neutral endopeptidase at 0.3 mg/kg/min, did not inhibit endothelin-converting enzyme or afford renal protection. The data suggest that the protective action of phosphoramidon against ischemic acute renal failure is most likely mediated by inhibition of endothelin formation. PMID- 8415870 TI - Effect of vasoactive intestinal peptide and naloxone combination on urinary N acetyl-beta-D-glucosaminidase level and kidney histology of rats exposed to severe hemorrhage. AB - Renal hypoperfusion which occurs in hemorrhagic shock creates an environment in which cellular injury and organ dysfunction can occur during the episode of shock as well as reoxygenation and reperfusion. At the same time, mast cell degranulation which is observed during hemorrhage may have an additional deleterious effect on the kidney. Twenty-two (Mus norvegicus albinos) rats (200 250 g) of either sex were used. The animals were divided into three groups. Group 1, the control group, was exposed to a 40% hemorrhage. Group 2 was exposed to 40% hemorrhage and then shed blood reperfused. Group 3 was exposed to 40% hemorrhage, and in addition to shed blood reperfusion 25 ng kg-1 vasoactive intestinal peptide (VIP) + 5 mg kg-1 naloxone (NLX) were given. At the end of the experiment the kidneys were evaluated either histologically or by measurement of the urinary N-acetyl-beta-D-glucosaminidase (NAG) activity. Shed blood reperfusion caused continuation of ischemic tissue damage and elevation of urinary NAG activity. Addition of VIP and NLX to the blood reperfusion caused a decrease in urinary NAG excretion, and the histology of renal tissue was almost normal. PMID- 8415871 TI - Carbon-tetrachloride-induced urinary excretion of formaldehyde, malondialdehyde, acetaldehyde and acetone in rats. AB - Previous studies have demonstrated that the hepatotoxin carbon tetrachloride rapidly promotes lipid peroxidation and inhibits microsomal calcium sequestration, microsomal glucose-6-phosphatase activity and cytochrome P-450. Due to its profound effects on lipid peroxidation, we have examined the oral administration of 2.5 ml/kg carbon tetrachloride on the urinary excretion of the lipid metabolites formaldehyde, malondialdehyde, acetaldehyde and acetone. Urine samples were collected up to 48 h after treatment. The urinary metabolites were identified and quantitated by gas chromatography-mass spectrometry and high pressure liquid chromatography. Time-dependent increases in the urinary excretion of the four metabolites were observed after carbon tetrachloride administration. At 48 h after treatment, the increases in the excretion of malondialdehyde, formaldehyde, acetaldehyde and acetone were approximately 55, 78, 57 and 268%, respectively, relative to control values. The data were expressed in nanomoles per kilogram body weight per 4.5 h. The results clearly demonstrate that carbon tetrachloride increases the urinary excretion of four lipid metabolites which may serve as noninvasive biomarkers of xenobiotic-induced lipid peroxidation. PMID- 8415872 TI - Neuromuscular transmission in nematode parasites and antinematodal drug action. AB - Some anthelmintic drugs interfere selectively with nematode neuromuscular transmission. These drugs include: the nicotinic agonists, e.g. levamisole, the gamma-amino butyric acid agonist piperazine, and the avermectins which open Cl- channels. The physiology and pharmacology of neuromuscular transmission in nematodes is reviewed and the actions of antinematodal drugs which interfere with the transmission described. The results of experiments on the large porcine intestinal nematode parasite, Ascaris suum, form the basis of the account presented but experiments on other nematodes suggest that these observations may be generalized. Results of some experiments on the small free living nematode Caenorhabditis elegans are also included. PMID- 8415873 TI - Organophosphate poisoning. AB - The present review discusses the structure of the anticholinesterase organophosphates (OPs), which are used predominantly as insecticides. OP poisoning can occur in a variety of situations and can be accidental or suicidal. It is common in developing countries. The cholinergic syndrome is caused by acetylcholinesterase inhibition, and diagnosis is based on the clinical signs and symptoms as well as the measurement of inhibition of erythrocyte acetylcholinesterase and/or plasma cholinesterase activity. Antidotal treatment is with atropine, an enzyme reactivator such as pralidoxime and diazepam. Anticholinesterase OPs may produce effects other than the acute cholinergic syndrome, including the intermediate syndrome. Later effects may include organophosphorus-induced delayed neuropathy. Certain OPs are exploited for their anticholinesterase effects, including defoliants such as 'DEF', herbicides such as glyphosate, fire retardants and industrial intermediates. The toxicology of this group is heterogeneous and they may or may not possess anticholinesterase activity. PMID- 8415874 TI - Liver cytoprotection by prostaglandins. AB - During the last decade intensive work on the relationships between the liver and the arachidonic acid cascade has greatly expanded our knowledge of this area of research. The liver has emerged as the major organ participating in the degradation and elimination of arachidonate products of systemic origin. The synthesis in the liver of arachidonate products derived from the cyclooxygenase, lipoxygenase and cytochrome P450 system pathways has been demonstrated. The participation of leukotriene B4 and cysteinyl-leukotrienes as mediators of liver damage and the possible therapeutic usefulness of prostaglandins (PGs) in acute liver injury has attracted the interest of clinicians. This article reviews the essential features regarding the role of arachidonate metabolites in liver disease and specially focuses on the cytoprotective effects on the liver displayed by PGE2, PGE1, PGI2 and synthetic PG analogs in experimental models of liver damage induced by ischemia-reperfusion injury, carbon tetrachloride, bacterial lipopolysaccharide and viral hepatitis and on the possible mechanisms underlying liver cytoprotection in these experimental models. The therapeutic usefulness of PGs in clinical practice is critically analyzed on the basis of available evidence in patients with fulminant hepatic failure and primary graft nonfunction following liver transplantation. PMID- 8415875 TI - Structural, functional and evolutionary implications of the three-dimensional crystal structure of murine interferon-beta. AB - The crystal structure of recombinant murine interferon-beta as elucidated by Senda et al. (Proc. Jap. Acad. 66B: 77-80 (1990); EMBO J. 11: 3193-3201 (1992)) appears to represent the basic structural framework of all Type I interferons including interferons-beta and all subtypes of interferons-alpha of various mammalian origin. Now the huge accumulated data on the structure-activity relationship of Type I interferons using various chemical and genetic techniques can be systematically evaluated in terms of the three-dimensional structure. Structural comparison with other cytokines, for which three-dimensional structures have been established, including interferon-gamma and considerations on the evolution of cytokines and cytokine receptors are also given. PMID- 8415876 TI - Mechanisms of halothane toxicity: novel insights. AB - Exposure of individuals to halothane causes, in 20% of patients, a mild form of hepatotoxicity. In contrast, a very small subset of individuals only develops halothane hepatitis, which is thought to have an immunological basis. Sera of halothane hepatitis patients contain antibodies directed against some discrete liver trifluoroacetyl (TFA)-protein adducts, which arise upon oxidative biotransformation of halothane and include protein disulfide isomerase, microsomal carboxylesterase, calreticulin, ERp72, GRP 78 and ERp99. No immune response occurs in the majority of human individuals, although evidence suggests that TFA-protein adducts arise in all halothane-exposed individuals. The lack of immunological responsiveness of individuals might be due to tolerance, induced by a presumed repertoire of self-peptides that molecularly mimic TFA-protein adducts. Thus, constitutively expressed proteins of 52 and 64 kDa have been identified that confer molecular mimicry of TFA-protein adducts. The 64 kDa protein corresponds to the E2 subunit of the mitochondrial pyruvate dehydrogenase complex. Lipoic acid, the prosthetic group of the E2 subunit, is involved in the molecular mimicry process. A fraction of halothane hepatitis patients exhibit irregularities in the expression levels of the 52 kDa protein and the E2 subunit protein. Molecular mimicry of TFA-protein adducts by the 52 kDa protein and the E2 subunit protein might play a role in the susceptibility of individuals to development of halothane hepatitis. PMID- 8415877 TI - Chemical disasters. AB - Chemical disasters are sufficiently common that plans are needed to prevent them, or to mitigate their effects should they occur. However, they are sufficiently rare that any individual is likely to have little or no experience of them. This article reviews chemical disasters which have been reported in the medical literature, and briefly discusses plans for medical involvement in the prevention and tackling of such events. PMID- 8415878 TI - Clinical pharmacology of corticosteroids in bronchial asthma. AB - It is now recognised that suppression of the inflammatory cascade should be the cornerstone of management in bronchial asthma. Inhaled corticosteroids are the most effective and widely used form of anti-inflammatory therapy for use in patients with asthma. The limited data available on dose-response relationships for inhaled corticosteroids suggest that a plateau occurs for antiasthmatic efficacy above 1600 micrograms either for budesonide and beclomethasone dipropionate with no appreciable differences between the two drugs. However, in most cases it should be possible to achieve adequate asthma control with doses of either drug less than 1000 micrograms with a use of an optimal inhaler device and good compliance. In contrast to topical anti-inflammatory activity in airways, for both local and systemic adverse effects there is a steep dose-response above 1600 micrograms with budesonide and beclomethasone dipropionate. In comparison with oral prednisolone there is still a better risk-benefit ratio even with higher doses of inhaled corticosteroids. There is evidence to suggest that inhaled budesonide may have a slightly more favourable profile in terms of the ratio of topical to systemic activity, particularly for effects on bone metabolism. A significant degree of adrenal suppression is unlikely at doses less than 1600 micrograms of budesonide or beclomethasone, although there is a degree of interindividual variability in the dose-response relationship for this effect, as well as for antiasthmatic activity. Thus, doses of inhaled corticosteroid should be titrated on an individual basis in order to achieve adequate disease control. At doses in excess of 800 micrograms it would seem rational to use a large volume spacer device since this will lessen local adverse effects such as oral candidiasis and dysphonia, as well as reducing systemic absorption and improving lung deposition. Mouth washing may reduce local and systemic adverse effects when using dry powder devices at high doses. Another possible strategy to improve efficacy with higher doses is to increase the dosing frequency from twice to four times daily, although this may at the same time produce an increase in local adverse effects. Fluticasone propionate is a novel inhaled corticosteroid with very high topical anti-inflammatory activity and minimal systemic bioavailability and might, therefore, provide a favourable therapeutic profile at the high end of the dose range. The next decade of research into the clinical pharmacology of inhaled corticosteroids is, therefore, eagerly awaited and will hopefully consolidate improvements in asthma management with this important class of anti-inflammatory drugs. PMID- 8415879 TI - Biological actions of oncogenes. AB - Cancer, in many cases, results from multistep genetic mutation. Certain genes can have a predisposed susceptibility to mutations that lead to cancer because of chromosome location or their importance in the control of cell cycles. Mutations that deregulate the expression or activity of enzymes involved in the biochemical pathways of growth and differentiation or that suppress the expression of negative cell cycle control factors result in activation of oncogenesis. The study of oncogenes and tumor suppressor genes has greatly influenced our understanding of the molecular origins of cancer. We focus here on the normal biological action of proto-oncogenes compared with the transforming activities of oncogenes and tumor suppressor genes, and we discuss possible mechanisms of oncogenic transformation. PMID- 8415880 TI - In utero drug therapy. AB - Drug therapy directed toward the fetus would be intended for either treating a fetal disorder or improving the capacity for later intrauterine or postnatal adaptation. Most reported trials involve single cases or small numbers of fetuses receiving the drug transplacentally after the first trimester, but before attaining maturity. Studies usually involve a single drug administered shortly before delivery. Treatments that are more direct or begun earlier in gestation are being attempted, but our limited understanding of fetal pharmacokinetics forces us to proceed cautiously. Studies to date have shown no risks to the mother and newborn infant, but long-term follow-up is necessary. PMID- 8415881 TI - Rhythms in the immune system. AB - This report reviews the evidence that cells within the immune system are subject to rhythmic influences that affect numbers of circulating cells and their function both in vitro and in vivo. It is concluded that, although periodicity has clearly been demonstrated for numbers of immunocompetent cells in the circulation, significant functional changes have not been consistently observed. A number of neuroendocrine hormones, which modulate immune responsiveness in vitro and which are released in a rhythmic manner, are considered as mediators of the observed effects on the immune system and this is related to changes in expression and activity of immune-mediated diseases such as rheumatoid arthritis. PMID- 8415882 TI - Health care research: old wine in new bottles. PMID- 8415883 TI - The new path to medical education at age twenty-one: the Kansas City experiment. PMID- 8415884 TI - AIDS in the 1990s: a global analysis. PMID- 8415885 TI - You can break your contract but not my heart. PMID- 8415886 TI - The healer's power: the dark side of the force--a conference. PMID- 8415887 TI - Relevance of John Donne to medical career preparation. PMID- 8415888 TI - Medical education and social needs. PMID- 8415889 TI - Questions and reflections. Be prepared. PMID- 8415890 TI - Dr. Ji replies to Dr. Brenner. PMID- 8415891 TI - Defensive medicine. PMID- 8415892 TI - Defensive medicine. PMID- 8415893 TI - Past and present. "The patient versus the costs". PMID- 8415894 TI - Pisse-prophets and puritans: Thomas Brian, uroscopy, and seventeenth-century English medicine. PMID- 8415895 TI - Serotonin syndrome-like symptoms in a patient with obsessive-compulsive disorder, following inappropriate increase in fluvoxamine dosage. PMID- 8415896 TI - EEG alterations and seizures during treatment with clozapine. A retrospective study of 283 patients. AB - In a retrospective study, 1863 EEG recordings made during clozapine treatment of 283 patients with normal pretreatment EEG evaluations were analyzed. Furthermore, they were compared to the EEGs of the same patients without clozapine (i.e., during other neuroleptic medication). Moreover, the data of all patients who had seizures during treatment with clozapine were evaluated in case reports. Classical clinical EEG evaluation criteria for normal versus abnormal were used (including diffuse slowing and grouped alterations according to Jung 1953 and Kugler 1983). Of the 283 patients investigated, 61.5% (174) showed at least one abnormal EEG under clozapine according to these criteria. Evaluating all recorded EEGs of these patients in order to get some longitudinal information, we found a rate of 53.4% abnormal EEG recordings during clozapine treatment. Most of the EEG changes were evaluated as slight (22.5%) to moderate (10.1%) diffuse slowing and some as groups of nonparoxysmal waves (39.8%) or sharp waves (16.2%) rendering the EEGs abnormal according to the above criteria. Potential signs of increased bioelectrical cerebral reagibility such as paroxysmal activity (4.3%) or severe diffuse slowing (0.2%) were rare. A nearly linear correlation with the daily dose was found in the range up to 300 mg clozapine/day for both diffuse and grouped alterations. Possibly due to selection, adaptive mechanisms/habituation, and/or other unknown factors, the rate of alterations decreased slightly at doses above 300 mg and rose again sharply for doses over 600 mg/d. Three of the clozapine treated patients, equivalent to 1.1%, developed seizures.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415897 TI - Double-blind multicenter study of paroxetine and amitriptyline in depressed inpatients. AB - Paroxetine is a new compound in the group of the selective serotonin-reuptake inhibitors. The results of several open and double-blind control-group studies demonstrate clear antidepressive efficacy of paroxetine. However, most data were collected in samples of outpatients. To overcome this restriction, a six-week double-blind control-group study, comparing 30 mg paroxetine with 150 mg amitriptyline per day, was performed in a sample of inpatients suffering from major depression. Generally speaking, the efficacy analysis of 160 patients was not able to demonstrate statistically significant differences in the antidepressive activity of paroxetine or amitriptyline, either with respect to the total score on the Hamilton Depression Scale (HAMD) and the Clinical Global Impressions or with respect to the subscores of the HAMD. One exception was the retardation subscore, in which amitriptyline showed a greater degree of reduction. Both drugs had a characteristic side-effect profile. Paroxetine was characterized by a lack of anticholinergic side-effects and a higher rate of nausea. PMID- 8415898 TI - Effects of low-dose dexamethasone on sleep EEG patterns, plasma cortisol, and the TSH response to TRH in major depression. AB - Dexamethasone (DEX) (0.5 mg, P.O.) and placebo were administered at 2300 h in randomized design to 19 patients with major depression and the effects on the sleep electroencephalogram (EEG) were studied. In addition, the thyroid stimulating hormone (TSH) response to thyrotropin releasing hormone (TRH) and basal plasma cortisol concentrations were assessed the following morning. DEX did not affect sleep architecture or continuity variables, including rapid eye movement (REM) latency, REM activity and REM density. Similarly, DEX did not significantly influence the TSH response to TRH (delta max TSH). In contrast, plasma cortisol concentrations were significantly suppressed by DEX. The results indicate that, as opposed to higher dosages of glucocorticoids, 0.5 mg DEX had minimal effects on the sleep EEG or delta max TSH in depressed patients. PMID- 8415899 TI - Adverse psychic reactions to psychotropic drugs--a report from the AMUP study. AB - The AMUP study (AMUP = Arzneimitteluberwachung in der Psychiatrie (Drug Monitoring in Psychiatry)) was conducted from 1979 to 1989 in order to provide for a systematic and standardized assessment of all adverse reactions to psychotropic drugs under the conditions of routine clinical treatment at two psychiatric hospitals. This paper presents data from the AMUP study on the type and frequency of adverse psychic reactions to psychotropic drug groups and relevant single drugs. Psychic ADR leading to drug discontinuation were observed in 4.5% of 15,264 inpatients monitored over an eight-year period. Only neurological ADR were more frequent (4.9%). Neuroleptics and antidepressants were involved with similar frequencies in ADR that were at least "probably" drug related (3.3 and 3.5%). Lithium salts and benzodiazepines were only rarely involved in psychic ADR. Toxic delirium (1.0%), agitation (0.9%), and sedation (0.8%) were the most frequent single events, usually rated as "probably" drug related. Depression and psychotic states were next in frequency, but judged as only "possibly" drug-related in a considerable proportion of cases. Haloperidol, the most common high-potency neuroleptic, was imputed mainly for depression, sedation, agitation, and (malignant) neuroleptic syndrome; with medium-potency perazine, toxic delirium and sedation prevailed; among the most common antidepressants, amitriptyline was above all connected with toxic delirium, while with clomipramine agitation predominated. The paper discusses the particular difficulties encountered in the field of psychic ADR in psychiatric patients regarding causality assessment, and emphasizes the need for continuous ADR assessment studies including state hospitals. PMID- 8415900 TI - Phospholipase A2 activity in serum of neuroleptic-naive psychiatric inpatients. AB - The activity of phospholipase A2 (PLA2), an enzyme that, besides a number of other functions, is involved in prostaglandin synthesis, regulation of membrane stability, and presynaptic neurotransmitter release, has been reported to be greater in the serum and plasma of schizophrenic patients than in that of other psychiatric patients and normal controls. Furthermore, it is said to be significantly reduced by haloperidol treatment. We decided to replicate these findings in an independent sample of patients by using a highly sensitive assay for determining PLA2 activity.-- METHODS: Of 167 consecutive male admissions giving informed consent 35 were found never to have received neuroleptics previously. Ten had DSM-III diagnoses of schizophrenia, schizophreniform disorder, or atypical psychosis, eight of other psychiatric disorders and 17 of alcohol or drug dependence (Table 1). Blood samples were drawn before medication was initiated. For patients still in hospital (Table 3), this was repeated after 21 days of treatment (n = 10). Samples from 10 healthy subjects were also obtained. PLA2 activity was determined according to Marki and Franson (1986), using a radiochemical assay.-- RESULTS: On day 0, there were no statistically significant intergroup differences in PLA2 serum activity. After 21 days, it was not possible to detect any significant decrease in PLA2 activity (Table 4). The PLA2 activity of the patient groups did not differ from that of the group of healthy subjects (Table 5).-- CONCLUSION: It was not possible to replicate the finding of increased PLA2 activity in schizophrenic patients in our sample of patients never treated with neuroleptics before.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415901 TI - Correction for photon attenuation in SPECT: analytical framework, average attenuation factors, and a new hybrid approach. AB - We provide a unified framework for the investigation of convergence properties of the iterative algorithms for photon attenuation correction in SPECT, including the iterative Chang method--a commonly used approach in which an average attenuation factor calculated over all projection angles is employed in a pointwise correction scheme. A new average attenuation factor calculated along the projection line is introduced, which can compensate exactly for the attenuation effect in the case of a uniform activity distribution. We propose a new hybrid approach in which we use this new average attenuation factor initially and then shift to the iterative Chang method in later iterations. This hybrid approach was evaluated in a simulation study by use of a computer-generated phantom with both non-uniform activity and non-uniform attenuation distributions. The results demonstrate that this hybrid approach improves the convergence speed of the iterative Chang method and produces reconstructed images of high quality. PMID- 8415902 TI - A new composite model of objects for Monte Carlo simulation of radiological imaging. AB - A composite object model is proposed for Monte Carlo simulation of radiological imaging systems. The composite model contains four components: a set of regular and 'voxelized' primitives, a 'modular' inclusion tree, a set of designated constructive solid geometry (CSG) trees, and a mapping from the set of CSG trees to the inclusion tree. The voxelized primitive is a primitive containing a stack of voxels whose intersections with a photon path are calculated based on Siddon's method. The inclusion tree is employed to describe the inclusion relationships of homogeneous subregions of material characteristics in larger regions in an object. The model is designed so that the 'divide-and-conquer' principle for modular software design can be used to construct an inclusion tree for a complex object. The designated CSG trees are used to model source distributions. The mapping from the inclusion tree to the CSG trees provides the fundamental information for the initial location of a photon history in the tree. Computational issues are addressed for the new model. For the modular inclusion tree, a computationally efficient algorithm is proposed in conjunction with the determination of the photon-ray intersections with primitive and voxel boundaries as well as with the identification of the material characteristics for each ray segment between two adjacent intersections. For source distributions, the designated CSG trees are defined based on the set intersections-difference-unions (IDU) operation sequence. The IDU operations present computation advantages over existing source modelling based on a single inclusion tree. For voxelized primitives, a new sampling technique called the fractional photon emission technique is introduced to reduce sampling computations for existing techniques based on single photon emission. The composite model is an extension of existing simple-geometry, solid-geometry, and voxel-based models and hence provides greater flexibility for model optimization. Based on the composite model, two objects were simulated: an anthropomorphic thorax with lesion inserts for transmission studies and a 3D Hoffman brain phantom with lesion inserts for emission studies. The simulations demonstrate the flexibility of the composite approach in modelling objects for clinically significant applications. PMID- 8415903 TI - A distributed quasi-static ionic current source in the 3-4 day old chicken embryo. AB - We report measurements of slowly varying magnetic field patterns close to fertilized eggs of the chicken Gallus domesticus during the first few days of incubation. These fields are generated by ionic currents within the egg that are associated with the development of the embryo. Since they are very weak (no greater than tens of pT) and vary over distances of a few millimetres, it has been necessary to develop specialized instrumentation and analysis techniques. We describe the use of high-spatial-resolution SQUID magnetometers to measure the field patterns and appropriate imaging algorithms to model the current sources responsible for producing the fields. Our results provide strong evidence for a distributed source in the extra-embryonic membranes. There is also indication of a more localized source within the embryo itself. PMID- 8415904 TI - Early skin reactions in head and neck malignancy treated by twice-daily fractionated radiotherapy--estimation of alpha/beta of LQ model. AB - Conventional radiotherapy, the five times a week regime, fails to achieve locoregional control in a large proportion of advanced carcinomas of head and neck and cervix. Different non-conventional fractionation schedules are being investigated to improve the tumour control rate but, during treatment of advanced malignancy with curative intent, it is seldom possible to deliver a radical dose to the tumour because of the constraint of inducing irreparable normal tissue complications. To compare the probability of occurrence of normal tissue damage and gain in therapeutic ratio various empirical models such as NSD, TDF, CRE and TSD have been suggested, and recently the linear quadratic (LQ) model has been gaining popularity and is claimed to be superior to earlier empirical models. We have studied twice-daily fractionated schedules in head and neck malignancy with the aim of estimating the alpha/beta value of the LQ model for early acute skin effects, and found it to be 9.16 Gy, 7.66 Gy and 8.59 Gy for mild erythema, intensive erythema and dry desquamation respectively. PMID- 8415905 TI - Adenosine-mediated photoreaction of 4,5',8-trimethylpsoralen with alcohols. AB - Irradiation of thin films consisting of 4,5',8-trimethylpsoralen (TMP), adenosine and small amounts of alcohols led to TMP-alcohol photoadducts in addition to TMP adenosine photoadducts. Four TMP-ethanol and two TMP-methanol adducts have been separated and characterized. Covalent bonds were formed between the 4-carbon of TMP and the alpha-carbon to the hydroxy group in the alcohols. The TMP-alcohol photoadducts were formed only in the TMP film containing small amounts of alcohol and adenosine. Furthermore, no photoadduct of TMP and ribose was detected upon photolysis of a TMP-ribose film, suggesting that the adenine moiety plays a specific role in the reaction. The interaction of adenosine with psoralens in a dry film may be related to the DNA sequence selectivity observed for the photoreaction of psoralens with DNA. PMID- 8415906 TI - Influence of hydration on the internal dynamics of hen egg white lysozyme in the dry state. AB - Proteins exist in a predominantly aqueous solvent environment. Hydration of the protein surface significantly affects many aspects of the protein's structure and function; these effects may be related to the molecular dynamics of the protein. We have examined the influence of hydration on the internal dynamics of hen egg white lysozyme using room-temperature phosphorescence from the intrinsic tryptophan residues. Powders of lyophilized lysozyme were hydrated in a phosphorimeter using a flow system that allowed for continuous manipulation of relative humidity over the range 0-92%; this system allowed us to directly compare intensity differences that result from changes in hydration. Lysozyme phosphorescence intensity decreased as a function of hydration over the entire relative humidity range; the decrease was not linear but appeared to occur in distinct phases. The phosphorescence intensity decays were multiexponential over the hydration range studied, and hydration had the largest influence on the long lifetime component. These data suggest that the protein exists in multiple, static conformations in the dry state and that water binding to polar (as opposed to charged) sites on the protein surface induces local and/or global softening of the protein structure. PMID- 8415907 TI - Electronic effects on the fluorescence of tyrosine in small peptides. AB - It is shown for a series of tyrosine-derivatives and tyrosine-containing peptides that the amide group in combination with electron-withdrawing substituents quenches the fluorescence of the phenol moiety. The ammonium group has the strongest electron-withdrawing effect and thus the largest influence on the quenching rate. The peptide group itself does not quench the fluorescence. In a series of peptides with an increasing number of alanines the decreasing quenching efficiency of the peptide group due to the greater distance of the ammonium group is demonstrated. In tyrosine-containing di- and tripeptides a linear correlation between the 13C-NMR chemical shift delta of the C alpha atom of various aliphatic amino acids and the fluorescence-quenching constant confirms the hypothesis that electron-withdrawing and -donating groups are modulating the fluorescence quenching efficiency of the peptide group. In small peptides the fluorescence lifetime of tyrosine is characteristic for the neighboring amino acids. Using model substances the redox properties of a peptide group and the phenol ring were studied electrochemically. The highest occupied molecular orbital of the tyrosine (1.4 V vs saturated calomel electrode [SCE]) and the lowest unoccupied molecular orbital of the peptide group (-3.12 V vs SCE) have appropriate energies for a photoinduced electron transfer reaction. For solute-quenching experiments quencher molecules can be systematically selected. PMID- 8415908 TI - Mechanisms of quenching of the fluorescence of a benzo[a]pyrene tetraol metabolite model compound by 2'-deoxynucleosides. AB - The hydrophobic interactions of bulky polycyclic aromatic hydrocarbons with nucleic acid bases and the formation of noncovalent complexes with DNA are important in the expressions of the mutagenic and carcinogenic potentials of this class of compounds. The fluorescence of the polycyclic aromatic residues can be employed as a probe of these interactions. In this work, the interactions of the (+)-trans stereoisomer of the tetraol 7,8,9,10 tetrahydroxytetrahydrobenzo[a]pyrene (BPT), a hydrolysis product of a highly mutagenic and carcinogenic diol epoxide derivative of benzo[a]pyrene, were studied with 2'-deoxynucleosides in aqueous solution by fluorescence and UV spectroscopic techniques. Ground-state complexes between BPT and the purine derivatives 2'-deoxyguanosine (dG), 2'-deoxyadenosine (dA), and 2'-deoxyinosine (dI) are formed with association constants in the range of approximately 40-130 M(-1). Complex formation with the pyrimidine derivatives 2'-deoxythymidine (dT), 2'-deoxycytidine (dC), and 2'-deoxyuridine (dU) is significantly weaker. Whereas dG is a strong quencher of the fluorescence of BPT by both static and dynamic mechanisms (dynamic quenching rate constant k(DYN) = [2.5 +/- 0.4] x 10(9) M( 1)s(-1), which is close to the estimated diffusion-controlled value of approximately 5 x 10(9) M(-1)s(-1), both dA and dI are weak quenchers and form fluorescence-emitting complexes with BPT. The pyrimidine derivatives dC, dU, and dT are efficient dynamic fluorescence quenchers (k(DYN) approximately [1.5-3.0] x 10(9) M (-1)s(-1), with a small static quenching component due to complex formation evident only in the case of dT. None of the four nucleosides dG, dA, dC and dT are dynamic quenchers of BPT in the triplet excited state; the observed lower yields of triplets are attributed to the quenching of single excited states of BPT by 2'-deoxynucleosides without passing through the triplet manifold of BPT. Possible fluorescence quenching mechanisms involving photoinduced electron transfer are discussed. The strong quenching of the fluorescence of BPT by dG, dC and dT accounts for the low fluorescence yields of BPT-native DNA and of pyrene DNA complexes. PMID- 8415909 TI - Fluorescence lifetime and quenching studies of sarcoplasmic reticulum Ca(2+) adenosine-5'-triphosphatase. AB - Different classes of tryptophan residues in sarcoplasmic reticulum calcium-ATPase were investigated with respect to their exposure to quenchers and sensitivity to high-affinity calcium binding to the ATPase. The charged quenchers, iodide and cesium, produced only slight quenching of ATPase fluorescence, whereas noncharged acrylamide and notably oxygen produced significant quenching. This finding gives support to the proposed location of most of the tryptophan residues of the ATPase in transmembrane domains of this protein (MacLennan et al., 1985, Nature 316r, 696-700). Among the different quenchers tested, oxygen quenching alone was sensitive to calcium binding to the ATPase, indicating that oxygen quenched tryptophan residues located in regions of the ATPase molecule which undergo conformational changes upon calcium binding. Time-resolved oxygen quenching data were analyzed with a recently described model that takes into account the existence of two different classes of emitters in the ATPase (Ferreira and Verjovski-Almeida, 1991, J. Lumin. 48, 430-434): a short-lived blue-shifted exponential component plus a long-lived red-shifted continuous lifetime distribution. Oxygen quenching of the single-exponential lifetime component was found to be insensitive to calcium, whereas quenching of the distributed lifetime component was significantly (ca 25%) enhanced by calcium binding. The different sensitivities of the two tryptophan classes to calcium binding to the ATPase are interpreted in terms of the proposed location of tryptophan residues in relation to the calcium transport sites in the ATPase molecule. PMID- 8415910 TI - Photosensitization with bacteriochlorins. AB - Biophysical and photobiological properties of a group of bacteriochlorins were compared with efficacy of these products for photodynamic therapy of murine tumors. Predictive factors for selective photosensitization in vivo include affinity binding to lipoproteins greater than albumin, extinction coefficient at the wavelength of irradiation and tumor/skin distribution. Efficacy was correlated with circulating plasma levels of the different sensitizers but not with the photodynamic therapy response in cell culture. PMID- 8415911 TI - Photodynamic effects of new silicon phthalocyanines: in vitro studies utilizing rat hepatic microsomes and human erythrocyte ghosts as model membrane sources. AB - Photodynamic therapy (PDT) of cancer is a modality that relies upon the irradiation of tumors with visible light following selective uptake of a photosensitizer by the tumor tissue. There is considerable emphasis to define new photosensitizers suitable for PDT of cancer. In this study we evaluated six phthalocyanines (Pc) for their photodynamic effects utilizing rat hepatic microsomes and human erythrocyte ghosts as model membrane sources. Of the newly synthesized Pc, two showed significant destruction of cytochrome P-450 and monooxygenase activities, and enhancement of lipid peroxidation, when added to microsomal suspension followed by irradiation with approximately 675 nm light. These two Pc named SiPc IV (HOSiPcOSi[CH3]2[CH2]3N[CH3]2) and SiPc V (HOSiPc OSi[CH3]2[CH2]3N[CH3]3+I-) showed dose-dependent photodestruction of cytochrome P 450 and monooxygenase activities in liver microsomes, and photoenhancement of lipid peroxidation, lipid hydroperoxide formation and lipid fluorescence in microsomes and erythrocyte ghosts. Compared to chloroaluminum phthalocyanine tetrasulfonate, SiPc IV and SiPc V produced far more pronounced photodynamic effects. Sodium azide, histidine, and 2,5-dimethylfuran, the quenchers of singlet oxygen, afforded highly significant protection against SiPc IV- and SiPc V mediated photodynamic effects. However, to a lesser extent, the quenchers of superoxide anion, hydrogen peroxide and hydroxyl radical also showed some protective effects. These results suggest that SiPc IV and SiPc V may be promising photosensitizers for the PDT of cancer. PMID- 8415912 TI - Alterations of proteoglycans in ultraviolet-irradiated skin. AB - The effect of UVB exposure on the distribution and synthesis of dermal proteoglycans was measured in the skin of hairless mice. Two groups of mice were included: one was irradiated for 10 weeks; the other was kept as control. After intraperitoneal injection of sodium 35-S-sulfate, punch biopsies were taken for histology and proteoglycans were extracted from the remaining skin with 4 M guanidinium chloride, containing 3-[(3-cholamidopropyl)dimethylammonio]-1 propanesulfonate (0.5%, weight per volume). Following proteolytic digestion, the glycosaminoglycan constituents were isolated and analyzed by quantitative cellulose acetate electrophoresis and enzymatic digestibility. Under the influence of UVB radiation, newly synthesized proteoglycans measured by 35SO4 uptake increased as much as 60%. In addition, the irradiated skin had a higher average content of proteoglycan than had control skin (4981 micrograms vs 4134 micrograms/g dry weight). This could be ascribed to an increase in heparin (1400 vs 533 micrograms/g dry weight) and heparan sulfate (472 vs 367 micrograms/g dry weight), whereas no change in the concentration of hyaluronic acid (1243 vs 1372 micrograms/g dry weight) and dermatan sulfate (1866 vs 1863 micrograms/g dry weight) was observed. The irradiated animals also exhibited a marked increase in the synthesis of heparan sulfate and heparin (62% and 71%, respectively). These results demonstrate that chronic doses of UVB altered proteoglycan metabolism through both quantitative and qualitative changes. PMID- 8415913 TI - In vivo repair of cytosine hydrates in DNA of cultured human lymphoblasts. AB - Ultraviolet irradiation of DNA in vitro results in the production of a wide variety of pyrimidine base alterations, including cytosine hydrates. Enzymes that initiate the repair of monomeric pyrimidine damage have been identified in both bacterial and mammalian systems; however, the in vivo formation and repair of cytosine photohydrates has not been demonstrated in cellular DNA. Using Escherichia coli endonuclease III as a damage-specific probe, we have shown that ring-saturated pyrimidines are formed in cultured human cells by irradiation with broad-spectrum UV light. In addition, these types of base damage are removed from the DNA of human lymphoblasts within 5 h following the irradiation. Analysis of the action spectrum for the formation of cytosine hydrates in DNA reveals that these photoproducts are formed most efficiently by irradiation in the range of 255-265 nm light, coinciding with the wavelengths that are maximally absorbed by the DNA bases. PMID- 8415914 TI - Activation of the human immunodeficiency virus promoter by UVA radiation in combination with psoralens or angelicins. AB - The effects of mono- and bifunctional furocoumarins plus UVA radiation (PUVA and related treatments) on the human immunodeficiency virus-1 (HIV-1) promoter were studied using HeLa cells stably transfected with the chloramphenicol acetyl transferase gene under the control of the HIV-1 promoter. The experiments were performed with three psoralens (5-methoxypsoralen, 5-MOP; 8-methoxypsoralen, 8 MOP; and 4'-aminomethyl-4,8,5'-trimethylpsoralen, AMT) and four angelicins (angelicin; 4,5'-dimethylangelicin, 4,5'-DMA; 6,4'-dimethylangelicin, 6,4'-DMA; and 4,6,4'-trimethylangelicin, TMA). The drugs alone and UVA radiation alone showed no effect on the HIV promoter. However, when the cells were incubated with the furocoumarins at 0.1-40 micrograms/mL and then irradiated, the HIV promoter was activated in distinct fluence ranges, i.e. (1) no promoter activity was discernible at low fluences (e.g. at 0.1 microgram/mL of 8-MOP up to 100 kJ/m2), (2) as the fluence was increased, the promoter activity increased to reach a maximum (10-50-fold with respect to the unexposed samples), and (3) as the fluence was further increased, the promoter activity decreased. Similar (although shifted on the fluence scale) patterns were observed with either > 340-nm UVA radiation or with UVA radiation contaminated with a small amount of UVB radiation (typical for PUVA lamps). The effective fluences were inversely related to the drug concentration. Experiments with 5-MOP and 8-MOP indicated reciprocity of the drug concentration and radiation fluence. The HIV promoter response patterns were similar for monofunctional angelicins and bifunctional psoralens.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8415915 TI - In vivo fluorescence kinetics of phthalocyanines in a skin-fold observation chamber model: role of central metal ion and degree of sulfonation. AB - The fluorescence pharmacokinetics of a series of metallosulfophthalocyanines, chelated with either aluminum or zinc and sulfonated to different degrees, was studied by fluorescence measurements in vivo. Dyes were administered systemically to female WAG/RIJ rats with an isogeneic mammary carcinoma transplanted into the subcutis in a transparent observation chamber located on their backs. Following an intravenous injection of 2.5 mumol/kg of the dye, fluorescence dynamics was observed up to 7 h postinjection. The phthalocyanines were excited at 610 nm with a power density of 0.1 mW/cm2 without causing photodynamic damage to the vasculature. Fluorescence was detected above 665 nm using a fluorescence imaging system based on an image intensifier. Dye retention in the blood vessels and tumor tissue was expressed as ratios relative to the fluorescence signal of the surrounding subcutaneous tissue. Phthalocyanines chelated with aluminum gave the highest fluorescence signal with tumor-over-subcutis ratios of up to a value of 4. The zinc complexes exhibited the highest vascular-over-subcutis ratios with maximum values exceeding a value of 6. They also displayed the longest retention times in the vascular system of well over 7 h. Overall, decreasing the degree of sulfonation of the metallophthalocyanines results in lower tumor-over-normal tissue fluorescence ratios, and furthermore aluminum-based dyes seem superior tumor localizers over zinc-based dyes. The advantages of phthalocyanines over porphyrins with respect to tumor localization and photodynamic therapy are discussed. PMID- 8415916 TI - 8-Methoxypsoralen DNA interstrand cross-linking of the ribosomal RNA genes in Tetrahymena thermophila. Distribution, repair and effect on rRNA synthesis. AB - The distribution and repair of 8-methoxypsoralen-DNA interstrand cross-links in the ribosomal RNA genes (rDNA) in Tetrahymena thermophila have been studied in vivo by Southern blot analysis. It is found that the cross-links at a density of < or = 1/2 x 10(4) base pairs (bp) are distributed equally between three domains (terminal spacer, transcribed region and central spacer) as defined by restriction enzyme analysis (BamHI and ClaI). It is furthermore shown that a dosage resulting in approximately one cross-link per rDNA molecule (21 kbp, two genes) is sufficient to block RNA synthesis. Finally, it is shown that the cross links in the rDNA molecules are repaired at equal rate in all three domains within 24 h and that RNA synthesis is partly restored during this repair period. The majority of the cells also go through one to two cell divisions in this period but do not survive. PMID- 8415917 TI - The anti-HIV activities of photoactive terthiophenes. AB - Various synthetic analogues of the naturally occurring terthiophene, alpha terthienyl (alpha T), were evaluated for anti-human immunodeficiency virus (HIV) activity. The compounds were incubated individually with a known amount of the virus, with or without UVA radiation (long-wavelength ultraviolet) and residual virus was monitored for its ability to produce cytopathic effects in cell culture and the production of virus-specific protein (p24). The basic terthiophene structure was essential for good anti-HIV activity, although various side chains, such as alcohols, bromo, methyl, thiomethyl and trimethylsilyl groups, permitted retention of maximum activity. Under optimum conditions, as little as 12 ng/mL of these compounds (i.e. approximately 3 x 10(-8) M) could inactivate 10(3) infectious virions. None of the compounds however were more active than alpha T itself. In all cases, UVA radiation was essential. Several side chains decreased the antiviral efficacy, and some side chains abrogated the activity. PMID- 8415918 TI - The effects of photodynamic therapy using differently substituted zinc phthalocyanines on vessel constriction, vessel leakage and tumor response. AB - The effects of four different zinc phthalocyanines were studied during and after photodynamic therapy (PDT). Measurements of vessel constriction, vessel leakage, tumor interstitial pressure, eicosanoid release, and tumor response of chondrosarcoma were made in Sprague-Dawley rats. Animals were injected intravenously with 1 mumol/kg of mono-, di-, or tetrasulfonated zinc phthalocyanine, or 1 mumol/kg of a zinc phthalocyanine substituted with four tertiary butyl groups. Tissues were exposed to 400 J/cm2 670 nm light 24 h after photosensitizer injection. An additional group of animals was given indomethacin before treatment. The use of the monosulfonated and tertiary butyl substituted zinc phthalocyanines in PDT caused the release of specific eicosanoids, caused vessel constriction, and induced venule leakage and increases in tumor interstitial pressure. Tumor cures of 27% and 7% were observed. Photodynamic therapy using the disulfonated zinc phthalocyanine did not induce vessel constriction or the release of eicosanoids, however, tumor cure was 43%. The use of the tetrasulfonated zinc phthalocyanine caused intermediate effects between the mono- and disulfonated compounds. The administration of indomethacin to animals completely inhibited the effects of PDT using the monosulfonated compound but had minimal effects on treatment using the disulfonated compound. This suggests that the monosulfonated and disulfonated compounds act by different mechanisms of destruction. PMID- 8415919 TI - Large mutagenic lesions are induced by photodynamic therapy in murine L5178Y lymphoblasts. AB - Mutagenic lesions at the thymidine kinase locus (tk) in mouse lymphoma L5178Y (LY) cells treated with red light and either Photofrin (PF) or chloroaluminum phthalocyanine (AlPc) as the photosensitizer were compared in the relatively photodynamic therapy (PDT)-sensitive strain LY-R16 and the relatively resistant strains LY-S1 and LY-SR1. Southern blot analysis revealed that 92% (36/39) of the PDT-induced thymidine kinase (TK-/-) mutants of strains LY-R16 and LY-SR1 lost the entire active tk allele. (Strain LY-S1 lacks a known tk polymorphism and has not been analyzed for loss of the active tk allele.) A decrease in galactokinase (GK) activity in the TK-/- mutants has been taken as an indication that the mutagenic lesion extends from the tk gene to the closely linked galactokinase gene (gk). Using PF as the photosensitizer, GK activity was decreased in 45% of the LY-R16 mutants and in 22% of the LY-S1 and LY-SR1 mutants. With photoactivated AlPc, 59% of the TK-/- mutants of strains LY-S1 and LY-SR1 showed GK inactivation. (LY-R16 mutants were not analyzed because of the low LY-R16 mutant frequency induced by PDT with AlPc) Thus, many of the TK-/- mutants of LY cells induced by PDT with either PF or A1Pc harbor multilocus lesions. PMID- 8415920 TI - General purpose lamps induce polyoma DNA replication in H3 cells. AB - We have previously demonstrated the ability of UVC (254 nm) radiation to induce asynchronous polyoma replication in rat fibroblast cells (H3 line) that contain an integrated copy of polyoma virus. In the present study we show that general purpose lamps can induce polyoma replication in these cells as well. The amount of UV radiation emitted by three different light sources was determined and the effects of each source on the replication of polyoma DNA was assessed. Our findings indicate that a 100 W incandescent lamp had a minimal effect on replication, whereas a 90 s exposure to a halogen lamp or a 160 W mercury vapor lamp induced replication 1.5-fold and 2-fold, respectively, in comparison with nontreated controls. We have previously shown that asynchronous polyoma replication in H3 cells involves UV-inducible cellular protein factors. Our present results indicate that these factors are also activated by exposure to commonly used lamps that emit comparable doses of UV radiation. PMID- 8415921 TI - A characterization of the fluorescent properties of circulating human eosinophils. AB - This paper presents a characterization of the fluorescence properties of human eosinophils isolated from peripheral blood of normal donors over a wide range of excitation and emission wavelengths. Circulating eosinophils possess three fluorescence excitation emission maxima: one at 280 nm excitation, 330 nm emission, attributable to tryptophan fluorescence, and currently unassigned peaks at 360 nm excitation, 440 nm emission and 380 nm excitation, 415 nm emission. Fluorescence microscopy studies show that the fluorescence of eosinophils may be site dependent; specifically, when observed at 365 nm excitation, circulating eosinophil fluorescence appears blue-violet, while the fluorescence of tissue dwelling eosinophils appears amber-gold. These results should be considered in developing an optical biopsy technique to identify eosinophils in human tissue. PMID- 8415922 TI - Evidence for the photoprotective effects of vitamin E. AB - The antioxidant vitamin E (alpha-tocopherol) may protect both animal and plant cell membranes from light-induced damage. The various biochemical and biophysical modes of protection are considered. An examination is made of the evidence that vitamin E plays an important prophylactic role against a number of serious light induced diseases and conditions of the eye (cataractogenesis and retinal photodeterioration) and skin (erythrocyte photohemolysis, photoerythema, photoaging and photocarcinogenesis) that are mediated by photooxidative damage to cell membranes. PMID- 8415923 TI - Protein serine/threonine phosphatases: structure, regulation, and functions in cell growth. AB - It is clear that much remains to be discovered regarding the roles of protein phosphatases in mitogenic signaling pathways. The ability of okadaic acid to activate MAPK/ERKs demonstrates that alteration in serine/threonine dephosphorylation can have significant effects on common steps in growth stimulation induced by different types of mitogens. As in the case of cell cycle control, protein serine/threonine phosphatase plays a central role in the reentry of quiescent cells into the cycle. Because the only known targets of okadaic acid are the catalytic subunits PP1 and PP2A, these enzymes are crucial components of two basic functions carried out by cells: growth and division. Important and obligatory roles for PP2B, PP2C, and newly discovered serine/threonine phosphatases are also likely. However, the limited tissue distribution, unique regulatory properties, and limited substrate specificities of these forms suggest more specialized functions in restricted cell types. The available information on the specific functions of different forms of protein serine/threonine phosphatases, let alone their individual isoforms and different multimeric holoenzymes, is still severely limited. Years of biochemical characterization and cDNA cloning have left us with far more forms than functions. This has led to the gratifying situation, at least for the biochemists, in which genetics and cell biology identify protein phosphatases for which a wealth of biochemical information is already available. The appreciation of the importance of these enzymes in the coming years can only increase as the functions for individual forms are discovered. PMID- 8415924 TI - Cholecystokinin and regulation of pancreatic acinar cell function. PMID- 8415925 TI - Mechanisms mediating renal secretion of organic anions and cations. PMID- 8415926 TI - Neutrophil signal transduction and activation of the respiratory burst. PMID- 8415927 TI - Gastroduodenal mucosal protection. AB - The barrier that protects the undamaged gastroduodenal mucosa from autodigestion by gastric juice is a dynamic multicomponent system. The major elements of this barrier are the adherent mucus gel layer, which is percolated by the HCO3- secretion from the underlying epithelial cells; the epithelial layer itself, which provides a permeability barrier and can rapidly repair superficial damage by a process of cell migration referred to as reepithelization or restitution; and a specially adapted vasculature, which provides a supply of HCO3- for transcellular transport and/or diffusion into the mucus layer. Passive diffusion of intestinal HCO3- into the lumen is particularly important when there is superficial damage resulting in increased leakiness of the mucosal epithelium. The process of reepithelization occurs by the migration of performed cells from gastric pits or duodenal crypts. This process is quite distinct from the wound healing and associated inflammatory response that accompany more severe injury or chronic damage. The adherent mucus gel acts as a physical barrier against luminal pepsin and provides a stable unstirred layer that supports surface neutralization of acid by mucosal HCO3-. Surface neutralization by mucosal HCO3- provides a major mechanism of protection against acid in the proximal duodenum. In the stomach, where luminal acidity can fall to around pH 1, other mechanisms of protection must exist, since the surface pH gradient is reported to collapse when luminal H+ exceeds approximately 10 mM. This collapse of the surface pH gradients may reflect, at least in part, that such studies have been mostly performed on non-acid-secreting mucosa where the supply of HCO3- to the interstitium from the parietal cells will be reduced. However, because the gastric mucosa can withstand prolonged exposure to acid without apparent damage, this implies an intrinsic resistance of the epithelial apical surface. This is amply illustrated within the gastric glands that do not secrete mucus and HCO3- yet are exposed to undiluted pepsin and an isotonic solution of HCl. Bicarbonate and mucus secretions together with mucosal blood flow are under paracrine, endocrine, and neural control. The rate of reepithelialization will depend on local chemotactic factors, adhesion mechanisms, and the creation of an acid/pepsin/irritant-free environment under a protective gelatinous or mucoid cap. If optimal conditions are met, then the rate of reepithelialization appears to depend primarily on the intrinsic properties of the migrating cells themselves rather than control by exogenous mediators.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8415929 TI - 1993 Ray G. Daggs Award. PMID- 8415928 TI - Scientific illiteracy: impact on research and what can be done. PMID- 8415930 TI - Signal Transduction and Gene Regulation, APS Conference. San Francisco, California, November 17-20, 1993. Abstracts. PMID- 8415931 TI - Effects of caffeine and gender on physiology and performance: further tests of a biobehavioral model. AB - The present study extended testing of our biobehavioral model describing the effects of arousal and caffeine to include an examination of gender differences and their interactions with habitual and acute caffeine ingestion. Males and females selected as high or low habitual caffeine users were randomly assigned to receive either caffeine or a placebo and exposed to novel and repetitive recall tasks and to simple auditory stimuli. Electrodermal activity and recall task performance were recorded. The four major factors examined, including habitual caffeine use, acute ingestion, gender, and stimulus novelty, affected behavior, physiology, or both. Results showed that habitual caffeine usage systematically affected tonic arousal (skin conductance level) and improved recall task performance. Acute caffeine ingestion increased phasic arousal (skin conductance response amplitude) and reduced habituation rates. Gender interacted with other factors to significantly affect both tonic and phasic arousal, and females performed better than males on the recall tasks. These results were partially supportive of the theoretical model, and further work is needed to examine the interactions of acute and chronic caffeine intake with gender and novelty. PMID- 8415932 TI - Amygdala kindling, anxiety, and corticotrophin releasing factor (CRF). AB - Wistar rats were kindled electrically in the anterior or posterior medial amygdala of the right hemisphere. One week after the fourth stage 5 seizure, anxiety was assessed in the elevated plus maze test. Anxiety levels of rats kindled in posterior medial amygdala were reduced relative to implanted controls, but not relative to unoperated controls. Kindling of the anterior medial amygdala increased anxiety relative to implanted and unoperated controls. The different effects of kindling on behavior were unrelated to any parameter of kindling. The stress of an ICV injection of saline increased anxiety in unkindled controls but reduced anxiety in anterior medial amygdala-kindled rats. Injection stress effects on behavior were blocked by 50 micrograms of alpha-helical CRF (the CRF receptor blocker). These findings suggest that CRF released by the stress of the injection procedure mediates the behavioral effects in both kindled and control rats. In contrast, injection of CRF (2 micrograms, ICV) has no greater effect than ICV saline in anterior medial amygdala kindled rats, whereas it was anxiogenic in unkindled rats. ICV vehicle and CRF reduce kindling-induced anxiety equally. These findings suggest that CRF released during the injection procedure saturates available CRF receptors. Finally, kindling did not alter basal plasma corticosterone levels. These and other findings suggest that the anxiety modulating actions of CRF are at central CRF receptors. PMID- 8415933 TI - Pain sensitivity in dietary-induced obese rats. AB - Previous literature indicates possible interrelationships between the endogenous opioids or endorphins, pain response, and obesity or eating behaviour. The pain response was, therefore, examined in a rat model of obesity induced by palatable food high in unsaturated fats. Pellet-fed control and energy-dense obese and nonobese rats were tested for latency of response to a thermal stimulus using the tail flick test. Obese rats exhibited a statistically significant increase in tail flick latency compared to controls. In addition, the observed latencies were significantly correlated to the body weight of the rats (r = 0.52, p < 0.01). These data suggest that dietary-induced obese rats are similar to obese humans in being less sensitive to painful stimuli, consistent with an increase in endogenous opioids in obesity. PMID- 8415934 TI - Satiety signals in sheep: involvement of CCK, propionate, and vagal CCK binding sites. AB - The satiety effects of the hormone, cholecystokinin (CCK), and propionate, an important gluconeogenic substrate, were studied in ad lib-fed sheep. Hepatic portal infusion of either sodium propionate (1.2 mmol/min) or sulphated CCK-8 (sCCK-8; 18.3 pmol/kg/min) had no effect on food intake. However, together, they decreased intake by 44%; similar to the effect of 2.4 mmol/min propionate alone. CCK infusions reduced the frequency of reticular contractions in the presence or absence of propionate. The effects of infusions on motility and food intake were, therefore, dissociated. Further studies demonstrated axonal transport of CCK binding sites in the ovine vagus. Binding sites accumulated to a similar extent on both sides of a ligature indicating the existence of both anterograde and retrograde transport which was limited to a small proportion of fibres. Binding incubations carried out in the presence of the CCK receptor antagonists, MK-329 and L-365,260, indicated that the majority of binding sites, if not the total population, possessed pharmacology typical of type B CCK receptors. PMID- 8415935 TI - Impairment of acquisition but not retention of a simple operant discrimination performance in aged Fischer 344 rats. AB - Although deterioration of learning and memory in aged rodents has been reported using a variety of tasks, little information is available on the effects of aging on acquisition and retention of reference memory in food-motivated tasks. In the present study, to examine reference memory, a simple operant discrimination task was used. Aged (24 month) and young (5 month) Fischer 344 rats were trained to discriminate brightness for food reward in an operant chamber. Aged rats showed significantly impaired learning compared to young rats, and the level of discrimination of aged rats was significantly lower than that of young rats. The aged rats also exhibited impaired learning in discriminating an auditory cue. Aged rats as well as young rats had good retention of brightness discrimination performance 30 days after the last training session. Scopolamine (0.3 mg/kg) did not impair discrimination learning nor affect well-trained discrimination performance in young rats. These data suggest that acquisition but not retention of reference memory is impaired in aged rats, and that the impaired acquisition observed in aged rats may not be due to dysfunction of the central cholinergic systems. PMID- 8415936 TI - Dopamine antagonists act on central, but not peripheral, receptors to inhibit sham and real feeding. AB - We examined the relative contribution of dopamine (DA) receptors in the brain and periphery in the control of sham and real feeding of sucrose solutions. Intraperitoneal (IP) administration of pimozide, an antagonist of peripheral and brain DA receptors, suppressed both sham and real feeding in a dose-related manner. In contrast, IP injections of domperidone, a DA antagonist restricted to peripheral receptors, had no effect on either sham or real feeding. The inability of domperidone to influence sucrose intake did not result from a lack of biological activity of the drug because the identical doses of domperidone that failed to alter eating significantly inhibited gastric acid secretion. The results implicate central, but not peripheral, DA receptors in the control of the ingestion of palatable foods and also suggest that sham feeding is more sensitive to DA antagonism than real feeding. PMID- 8415937 TI - Proceedings of the First International Behavioral Neuroscience Conference. San Antonio, Texas, May 21-24, 1992. PMID- 8415938 TI - Activin A: serum levels and immunohistochemical brain localization in rats given diets deficient in L-lysine or protein. AB - When a L-lysine (Lys)-deficient diet is given to rats, Lys in plasma and brain declines and rats will then select a Lys solution from among other L-amino acids (AAs). The recording of single-unit activity in the lateral hypothalamic area of these rats suggested that neural plasticity occurred, specifically responding to the deficient nutrient, Lys, centrally and during ingestion of AA. Possible neurotrophic factors in serum from rats with or without deficiency of either protein or Lys was assayed by Hydra japonica. An increase in serum inhibin and activin A was observed in rats fed a Lys-sufficient and nonprotein diet, respectively. However, serum activin A-like activity was severely suppressed under Lys deficiency. Additionally, the immunohistochemical distribution of activin A in the brain was found in the nucleus tractus solitarius, the area postrema, and the arcuate nucleus. These facts indicate that ingestion of Lys deficient or nonprotein diet caused a change in serum levels of activin A as a possible neurotrophic factor. This release may elicit plasticity in the sensitivity of neurons to deficient AA in the nuclei that could selectively drive ingestive behavior for its particular AA (e.g., Lys) to maintain AA homeostasis. PMID- 8415939 TI - Abnormal weight gain in rats with amygdaloid lesions. AB - Marked weight gain was observed in female rats given small electrolytic lesions in the dorsal posterior portion of the amygdala. With a standard lab pellet diet, weight gains typically ranged between 20-30 g during the first 3 postoperative days, and between 60-100 g over the first 20 days. Rats with sham lesions generally gained only 5-15 g in 20 days. The results are consistent with much older studies that reported obesity in cats, dogs, and primates with lesions of the amygdala. PMID- 8415940 TI - Olfactory bulbectomy in rats modulates feeding pattern but not total food intake. AB - The role of olfactory input in the regulation of food intake and feeding patterns in rats was investigated by performing bilateral olfactory bulbectomy. Compared to control rats, bulbectomized rats ate the same amounts of food, but did so via a decrease in meal size, and a doubling in meal number. Although no increase in meal duration occurred, the exploratory behavior of sniffing during meals and between meals also increased significantly. While it is not yet clear how the olfactory bulbs participate in regulating food intake configuration resulting in changed feeding patterns, their clinical role can be appreciated by observing the acute changes in feeding pattern that occur when their input to the lateral hypothalamic area is damaged experimentally. PMID- 8415941 TI - Insulin and satiety from feeding in pancreatic-normal and diabetic rats. AB - Insulin's role in food ingestion and satiety was investigated in streptozotocin diabetic and pancreatic-normal rats. Markedly diabetic rats (60-65 mg/kg b.wt. streptozotocin with mean glycemia of 380 mg/dl) were observed for daily food/water intake and body weight changes and meal pattern in a standard operant chamber. Diabetic animals showed an immediate hypophagia (days 1-4) by decreasing meal size (MS). As animals lost weight, a significant hyperphagia appeared, accomplished by an elevation in MS. Treatment with insulin infused via osmotic minipump at a stable rate (6.0 U/24 h) reduced polyuria and glycemia, eliminated glycosuria, and restored weight gain. MS did not, however, return to baseline immediately. In a second experiment with milder diabetes (20-22 mg/rat streptozotocin, which produced glycemia ranging from 148 to 304 mg/dl), significant hyperphagia appeared, again attributable to an increase in MS. Minipumps infusing 2.4-4.0 U insulin/24 h reversed this hyperphagia. In pancreatic-normal rats, pumps infusing insulin at 1.2 or 2.4 U/24 h produced a modest hypophagia accomplished by a primary decrease in nocturnal intake (11% suppression in dark feeding). Subdiaphragmatically vagotomized rats showed an attenuation of this suppression (no significant effects on food or water intake produced by insulin infused). Insulin appears to participate in satiety by limiting MS, without significantly shortening latency to take the next meal. PMID- 8415942 TI - Abnormalities in obese Zuckers: defective control of histaminergic functions. AB - Histaminergic functions in the hypothalamus of Zucker obese rats were investigated. Blockade of postsynaptic H1-receptor after infusion of chlorpheniramine into the third cerebroventricle (ICV) failed to affect feeding in obese Zuckers, although feeding was potently elicited in Wistar King A control rats. Presynaptic increase in histamine by an H3-receptor antagonist, thioperamide, suppressed feeding in Wistar controls, but not in obese Zuckers. Under high ambient temperature, Wistar controls decreased food intake and maintained their rectal temperature normally. However, obese Zuckers and histamine depleted rats due to alpha-fluoromethyl-histidine (FMH), a specific "suicide" inhibitor of a histamine synthesizing decarboxylase enzyme (HDC), failed to show this decrease in food intake as adaptive behavior. Their rectal temperature concomitantly elevated in response to heated circumstance. ICV infusion of thioperamide increased the blood glucose level in Wistar controls, but not in obese Zuckers. The defect in all these regulatory functions found in obese Zuckers may be derived from an excessive decrease in hypothalamic histamine content due to inactivity of HDC. The histamine-depleted model sufficiently mimicked the abnormalities in obese Zuckers. PMID- 8415943 TI - Effects of chronic stress and time of day on preference for sucrose. AB - Rats were allowed to lever press for two types of food pellets of equivalent caloric value and total carbohydrate content. One type of food pellet provided more of the calories as sucrose. During a 16-day prestress period, lever presses for 12 rats were recorded hourly. Following the baseline period, four rats (stressed group) were shaped to pull a ceiling chain to avoid or escape signalled foot shock presented intermittently around-the-clock. Four additional rats (yoked group) were each paired to one of the chain-pulling rats such that the rat trained to pull the ceiling chain controlled stressor termination for both rats. A third group of four rats served as the control group and received no shock. We have previously reported that rats in this model of chronic stress tolerate the paradigm well, continuing to gain weight, eat, drink water, and groom and escape more than 99% of the trials presented. During the baseline period, the sweeter pellet was preferred by most rats, but differences in preference among rats and in preference at different times of day were observed. The preference for the high-sucrose pellet was most marked in the hours preceding lights off. Overall, no changes in food preference were seen as a function of stress condition during the 14-day stress period, although one rat in the yoked group increased preference for the sweeter pellet during stress and returned to prestress food preferences when stress was terminated. PMID- 8415944 TI - Mid-life onset of dietary restriction extends life and prolongs cognitive functioning. AB - Fourteen-month-old C57BL/6 (NIA) mice were placed on a nutritionally complete diet providing 139.4 kcal/week. Over a 2-month period the food ration of experimental mice (AE) was reduced to 85 kcal/week, where it remained for the duration of the study. An aged control group (AC) continued with the higher calorie diet. At age 22 months, AC mice and half of the AE mice (AE22) were given a battery of behavioral tests. The remaining AE mice (AE25) were given the test battery at age 25 months. Also, a middle-aged control group (MC) was tested at age 13 months. Midlife onset caloric restriction (CR) increased longevity and preserved strength, coordination, and spontaneous alternation behavior, and altered responses to enclosed alleys. A spatial discrimination in the Morris water maze and a spatial delayed matching-to-sample water-escape task were insensitive to age and diet. The aged mice were adversely affected by testing. PMID- 8415945 TI - The effects of cocaine on dietary self-selection in female rats. AB - Cocaine was administered via an oral route to 18-h food deprived female rats for 14 consecutive days. Following administration of the drug or vehicle control each animal was presented with separate isocaloric rations of protein, fat, and carbohydrate in a dietary self-selection situation. Amounts consumed of each component were measured at 30 min, 60 min, 2 h, and 6 h following the drug treatment. The intake of all three macronutrients was suppressed by cocaine for 1 h. Between 2 and 6 h after administration, there was a compensatory increase in fat and carbohydrate, but not protein consumption. The results are discussed in terms of protein deficiency caused by cocaine in pregnant and/or lactating females being a causal factor in the deleterious effects on offspring. PMID- 8415946 TI - Effects of phenylpropanolamine on regulatory and nonregulatory ingestion in adult rats. AB - This experiment examined the effects of phenylpropanolamine (0.0, 5.0, 10.0, 20.0 mg/kg PPA) on regulatory (RG) and nonregulatory (NRG) eating and drinking in rats using a within-subjects design. Administration of PPA produced dose-dependent reductions in eating in animals deprived to 80-85% of baseline weight, and reduced drinking after 23.5-h of water deprivation. Nonregulatory eating, elicited by tail pinch in nondeprived animals, was similarly inhibited. Nonregulatory drinking was elicited in the schedule-induced polydipsia (SIP) paradigm. Water consumption, locomotion, licking, lick efficiency (licks/ml water), and entries into the food magazine were simultaneously measured. At the lowest dose, only locomotion was significantly reduced. At 10.0 mg/kg, lick efficiency and entries into the food magazine were also significantly reduced, while all measured behaviors, including licking and water consumption, were decreased by the highest dose of PPA. The reduction in lick efficiency suggested a PPA-induced motor impairment in the capacity for licking. Considered together, these results indicated that the observed decreases in regulatory and nonregulatory eating and drinking could be at least partially accounted for by the drug's effects on behaviors contributing to ingestion, as well as apparent motor impairments in ingestive behavior at higher doses. PMID- 8415947 TI - Effects of phenylpropanolamine infusion and withdrawal on body weight and dietary composition in male and female rats. AB - Male and female rats with ad lib access to separate sources of carbohydrate, fat, and protein were implanted with minipumps providing one of three dosages (0.0, 40.0, or 80.0 mg/kg/day) of phenylpropanolamine (PPA) for 2 weeks. Body weight, macronutrient intake, and water consumption were measured daily before, during, and after PPA treatment. Phenylpropanolamine lowered body weight and caloric intake in males and females, and water consumption in females, but did not alter dietary composition in either sex. After PPA termination, caloric intake returned to control levels in both males and females. However, body weight returned to control levels in males only, while PPA-treated females continued to weigh less than controls. Phenylpropanolamine termination was associated with significant increases in water consumption and the percentage of total calories consumed from protein and reductions in the percentage of calories from carbohydrate in males. In contrast, water and macronutrient consumption was similar comparing PPA treated females to controls after drug termination. These results suggest there are sex differences in the effects of PPA termination on water and macronutrient consumption that result in differential weight gain in males and females. PMID- 8415948 TI - Partial limbic kindling--brain, behavior, and the benzodiazepine receptor. AB - Partial kindling (PK) of the left perforant path (PP) lastingly increased feline defensiveness. Perforant path PK produced long-term potentiation (LTP) in the amygdalo-ventromedial hypothalamic (AM-VMH) pathways in both hemispheres, and in the ventroamygdalofugal (VAF)-VMH efferents of the amygdala of the left hemisphere. Long-term potentiation paralleled behavioral changes. Perforant path PK did not affect recurrent inhibition in area CA3 of the ventral hippocampus. Long-term potentiation of CA3 EPSP and population spikes appeared, but before behavioral changes. Changes in excitability of the periaqueductal grey also accompanied behavioral changes. After kindling, the benzodiazepine receptor antagonist, flumazenil, reduced defensive response to rats in a drug-dependent manner. Flumazenil also reduced LTP in the AM-VMH pathway, but did not affect LTP in the VAF-VMH pathway. Therefore, flumazenil was acting in the amygdala, and not at the VAF-VMH synapse. Kindling caused flumazenil to act like an agonist on behavior, and in the amygdala, and as an agonist or an inverse agonist in area CA3 depending upon the physiological measure taken. PMID- 8415949 TI - Behavioral and electrophysiological comparison of ketamine with dizocilpine in the rat. AB - We have compared the effects of MK 801 and ketamine on a measure of anesthesia (loss of righting reflex) and two measures of basal ganglia dopamine (DA) function: apomorphine (APO)-induced stereotypy and APO-induced excitation of type II globus pallidus (GP) neurons. As expected, ketamine induced anesthesia. High dose MK 801 administered IP induced ataxia, but not anesthesia. When administered i.v., high-dose MK 801 induced anesthesia in only three of five rats. Using a modified stereotypy scale, it was found that pretreatment with MK 801 blocked APO induced stereotypic sniffing. Intravenous ketamine also blocked APO-induced stereotypy, but IP ketamine did not. Similar results were observed in neurophysiological studies; MK 801 altered the excitation of type II GP neurons by APO. Intravenous ketamine (5 mg/kg) also altered the responsiveness of these cells to APO, but ketamine anesthesia (150 mg/kg, IP) had no effect. These findings suggest that MK 801 is not an effective anesthetic in rats, and the method of administration of ketamine plays a role in its ability to exert NMDA receptor blockade. PMID- 8415950 TI - Effects of light stimulation on the activity of the autonomic nerves in anesthetized rats. AB - The existence of the retino-hypothalamic pathway suggests that light stimulation may influence the activity of the autonomic outflows. Efferent activities of the pancreatic, hepatic, and gastric branches of the vagus nerve and those of pancreatic, hepatic, splenic, adrenal, and renal branch of the splanchnic nerve were recorded. Light stimulation with 2000 1x for 10 min to the left eye increased the splanchnic (sympathetic) outflows and suppressed the vagal outflows. The effects lasted for several hours. The minimal effective stimulation was 20 1x for 1 min or 200 1x for 0.1 min. These responses were observed in the light period as well as dark period. However, in the suprachiasmatic nucleus (SCN) lesioned rat, the changes in autonomic outflows following light stimulation were absent. The observations suggest that light stimulation modulates visceral functions through changes in the autonomic nervous system activities via the SCN. PMID- 8415951 TI - NMDA receptors modulate long-term habituation to spatial novelty: dose- and genotype-dependent differential effects of posttrial MK-801 and CPP in rats. AB - To investigate the role N-methyl-D-aspartate (NMDA) receptors play in behavioral plasticity, adult male rats of the Naples high-(NHE) and low-excitability (NLE) lines, and of a random-bred Sprague-Dawley strain (NRB) received, the noncompetitive (MK-801:0.01 or 2.5 mg/kg) or the competitive (CPP: 0.01 or 5 mg/kg) NMDA receptor antagonists, or vehicle IP soon after a 10-min test in a Lat maze. Retention was tested 1 week later. Habituation of activity and defecation score was monitored by the between-test decrement (LTH) in the frequency of corner-crossings (HA) and rearings (VA), with prevailing cognitive and noncognitive meaning, respectively, and of fecal boli. (i) In the NLE-rats, low and high doses of MK-801 facilitate LTH of HA, and a high dose of CPP facilitates LTH of HA. (ii) In the NRB-rats, MK-801 facilitates LTH of HA at a low dose and inhibits LTH of VA at a high dose, whereas CPP inhibits LTH of HA at a high dose only. In contrast, (iii) in the NHE-rats, high doses of MK-801 impair LTH of HA, and low doses of CPP facilitate LTH of HA. In conclusion, the dose- and genotype dependent differential effects of allosteric and isosteric receptor blockade support the hypothesized modulatory role of NMDA receptors in behavioral plasticity; and the dissociation between retention of cognitive and noncognitive behavioral components suggests that NMDA receptors are involved in their parallel processing. PMID- 8415952 TI - Nimodipine's functional benefits depend on lesion completeness in medial septal area. AB - The effects of a 4-day nimodipine treatment (70 micrograms/kg IP beginning on the day of surgery) given to rats with lesions directed at the medial septal area were monitored for 120 days. Body weight, water intake, open-field activity, rearing, hole-poking, and repetitive motor acts were periodically measured through 120 postsurgical days. Although no differences were found in water intake between any of the groups, the body weights of rats with any medial septal damage, whether treated with nimodipine or not, were lower than rats with control operations by postsurgery day 120. Rats with any medial septal damage, whether treated with nimodipine or not, had lower rearing frequencies, rearing durations, and hole-poking frequencies than controls on all test days. However, rats with complete medial septal lesions treated with nimodipine exhibited movement in the open field and frequencies of stereotyped, species-typical acts similar to those of control rats by postsurgery day 60. This nimodipine effect was not observed in rats with partial lesions of the medial septal region. This study emphasizes that a brief administration of nimodipine shortly after brain damage can influence behavioral changes 40-60 days after surgery, but that this effect was not apparent in rats with only partial medial lesions. PMID- 8415953 TI - An analog technique for recording leg position learning in the cockroach. AB - Past studies of shock avoidance learning in the cockroach have used an all-or none method of recording leg position. Leg position was recorded indirectly in terms of the number of shocks the leg initiated. The measure of learning was the decrease in the number of times the leg initiated shock. An analog procedure is described that allows the pattern of leg behavior change to be assessed directly. Thus, the time course and relative magnitude of leg flexions and extensions prior to, during, and following shock can be studied. It will be possible to record peripheral motor nerve activity simultaneously with the behavior and to examine the excitatory and inhibitory interactions of individual neurons that may be involved in such learning. PMID- 8415954 TI - Transplant-induced working memory deficits in hippocampectomized rats. AB - This experiment determined the effects of transplantation of fetal hippocampus on the ability of male rats with hippocampal lesions to acquire versions of a radial arm maze that depended on either extramaze cues or intramaze cues for solution. Rats receiving transplants took significantly more trials than control rats to emit three consecutive errorless trials in the extramaze cue (spatial) variation of the maze. Rats with just hippocampal lesions never differed from any other group. No differences in this measure were found for the intramaze cue condition. Rats receiving transplants made more repeat entries into reinforced arms in both versions of the maze than control rats and more reentries into neverbaited arms in the spatial maze. Rats with hippocampal lesions failed to differ from any other group on this measure in the spatial maze, but were different from normal rats in the intramaze cue maze. These data suggest that in some tasks transplants of fetal tissue lead to greater behavioral impairment than lesions alone. PMID- 8415955 TI - Reduction of motor impairment by adrenal medulla transplants in aged rats. AB - Recently we have reported that drugs that enhance dopaminergic transmission, such as L-dopa and D-amphetamine, substantially improve the age-related deterioration of extrapyramidal motor functions, as assessed by the narrow uphill beam test. Here we report the effect of fetal adrenal medullary transplants upon the motor performance of aged rats in such a test. Rats were grafted with 300,000 cultured adrenal medullary cells, placed into the head of either the left caudate nucleus, or into both caudates. A third group was grafted with one freshly dissected adrenal medulla placed as a block of tissue into the lateral ventricle, whereas the control group sustained sham grafting. Evaluation of the motor performance of cultured and sham-grafted rats showed no improvement along the testing phase. Only adrenal block-grafted rats exhibited a significant recovery of motor coordination. Histologically, cultured cell grafts had a deteriorated appearance with poor survival rates, while block grafts exhibited chromaffin cells with round and neuron-like shapes. The results suggest that cultured adrenal grafts may not induce motor improvement due to their extremely low survival and poor integration, at least in the aged host brain, while fresh adrenal transplants may improve motor coordination for as long as 84 days. PMID- 8415956 TI - Heavy water lengthens the period of free-running rhythms in lesioned hamsters bearing SCN grafts. AB - Heavy water (D2O) lengthens the period of free-running circadian rhythms in most organisms. We compared the effect of D2O on free-running locomotor activity rhythms in intact and SCN-lesioned (SCN-X) hamsters that had recovered circadian rhythmicity following implantation of SCN grafts. The animals were housed individually in cages equipped with running wheels, and locomotor activity was monitored using a computer-based data acquisition system. At the end of the behavioral tests, animals were anesthetized and perfused. Brain sections were immunostained for vasoactive intestinal peptide (VIP) and vasopressin (VP) to evaluate the extent of the lesion and the presence of a functional graft. The D2O similarly lengthened the period of free-running activity without affecting amount of activity in both intact and in SCN-X grafted animals. The results indicate that D2O acts directly on the SCN to lengthen the free-running period, and suggest that coupling between pacemakers within the grafted SCN is as efficient as in the intact SCN. PMID- 8415957 TI - Functional mapping of the rat brain during drinking behavior: a fluorodeoxyglucose study. AB - Autoradiographic techniques using the radiolabeled glucose analog [14C]2-fluoro-2 deoxy-D-glucose (FDG) were used to map the functional activity in the CNS during drinking behavior. Rats were trained to drink water during a 1-h session each day. Half of the rats were injected with FDG and allowed to drink, while the other half were satiated prior to FDG injection. Uptake of FDG for drinking and control groups of rats was quantified in 60 brain structures from frontal cortex to cervical spinal cord. The largest percent increase in activity (96%) during drinking was in the lateral hypothalamus. Limbic structures with significant metabolic increases included the lateral septum (48%), lateral habenula (44%), and nucleus accumbens (32%). Thalamic nuclei activated included intralaminar (60%), zona incerta (51%), ventroposteromedial (50%), anterior ventral (47%), and dorsal medial (40%). Other structures with increases were the caudal caudate nucleus (53%) and the spinal trigeminal nucleus (45%). The findings were interpreted in light of related metabolic mapping studies of the effects of orofacial stimulation, dehydration, ingestion, arousal, and reward. It was concluded that this FDG study revealed primarily the involvement of structures linked to rewarding and arousal components of motivated drinking behavior, as well as sensorimotor correlates of the orofacial stimulation. The findings provide the first comprehensive functional map of brain systems related to drinking behavior in adult animals. PMID- 8415958 TI - The impact of the media on women with breast implants. AB - Because of the considerable attention given to breast implants by the media, these two surveys were conducted to evaluate how women with breast implants think and feel about the current issues. A telephone questionnaire was administered to 60 women prior to the Food and Drug Administration's moratorium and again following the advisory panel recommendation. The second survey revealed that most subjects felt the media information was accurate, a significant (p = 0.02) change from the first survey responses. The most frequently cited concern about breast implants on the first survey was silicone leakage. It continued to be a major concern on the follow-up survey but was replaced by mammography concerns in the women with augmentation. Concern regarding autoimmune disease increased significantly on the follow-up survey (p = 0.0001). Satisfaction with the decision to have implants changed from 98 percent of the sample before the moratorium to 71 percent of augmentation patients and 79 percent of reconstruction patients after. Women willing to choose the procedure again also declined from 72 percent of reconstructive patients and 58 percent of augmentation patients before to 59 and 52 percent, respectively, after (p = 0.01). The majority (67 percent) acknowledged that the moratorium had an effect on their feelings about breast implants. Negative media information appears to have been a persuasive influence on patients' opinions regarding their implants. PMID- 8415959 TI - Breast reconstruction utilizing implants: a local experience and comparison of three techniques. AB - The breast reconstruction patient requires accurate information about the complications, failures, and additional surgeries associated with implant reconstruction. In this series of consecutive patients, we examine three methods of implant reconstruction: (1) immediate with a permanent implant/expander (86 patients/107 breasts), (2) delayed with a permanent implant/expander (57/73), and (3) delayed serial expansion with gel/saline (54/68). Implant/expander reconstructions had an equal interval to completion (immediate 118 days, delayed 127 days), a significant need for revisions (immediate 57 percent, delayed 30 percent), and a similar complication rate (10 percent) and failure rate (3.5 percent). The delayed serial expansion patients were completed with one operation 66 percent of the time, and only 9 percent required revisional surgery. The data obtained from the largest private health care provider in the state confirmed a significant difference (p = 0.003) in the need for revisional surgery between immediate and delayed reconstructions. The data showed immediate implant reconstructions to (1) be safe (low failure rate), (2) require more capsular intervention procedures, (3) have a greater expense, and (4) have good aesthetic results (90 percent Baker class I or II) because of revisional surgery. The immediate implant breast reconstruction patients are committed to their reconstruction and undergo revisional surgery to improve their aesthetic result. Delayed implant reconstruction patients are less likely to undergo further surgery to correct capsular deformities. PMID- 8415960 TI - Breast cancer diagnosis and survival in women with and without breast implants. AB - The stage at diagnosis and the survival experience of 41 women who developed breast cancer after cosmetic breast augmentation were compared with those of all other patients with breast cancer (n = 13,246) diagnosed in Alberta from 1973 to 1990 (inclusive). The tumors in women with breast implants were smaller (65.9 percent < or = 2 cm) as compared with the tumors in women without implants (34.1 percent < or = 2 cm), but lymph node and distant metastases were equally frequent in the two groups. The distribution of tumor histologic types did not differ significantly between women with or without implants. Women who had an implant were younger at diagnosis of breast cancer compared with women with breast cancer and no breast implants. The relative 5- and 10-year survival rates did not differ significantly between the two groups, and the Kaplan-Meier survival estimate also was similar. It is concluded that women with breast implants in whom breast cancer develops are not diagnosed in a later stage and do not experience an impaired survival as compared with breast cancer patients without implants. PMID- 8415961 TI - The endoscopic breast augmentation: the transumbilical insertion of saline-filled breast implants. AB - Initial aesthetic breast augmentation with inflatable saline implants has always had the distinct advantage of insertion through a small breast or axillary skin incision. Adapting established techniques utilized in suction-assisted lipectomy, operative endoscopy, and tissue expansion with inflatable saline implants, breast augmentation is possible through an umbilical incision. Under general anesthesia, an incision is made in the umbilicus; a new tubular instrument with an obturator [designated an endotube (Johnson) or mammascope (Christ)] is inserted into the umbilical incision like a suction cannula; it is pushed over the abdominal fascia across the costal margin until it literally pops under the breast fascia; through this tunnel is then inserted an implant coiled like a tobacco leaf; the implant is then inflated to 50 percent more than the final volume and manipulated to help expand the pocket; finally, the excess volume is removed, methyl-prednisolone acetate is placed in the final volume, and the fill tube is removed. The endoscope (laparoscope) is utilized to visualize positioning and to document the absence of bleeding. The umbilical incision is closed after insertion of both implants through the same incision. A series of 91 young women have undergone this procedure with 188 breast implantations without significant bleeding. Implants appear to ride high initially, but they settle into place by 6 weeks. Patients have reported less chest discomfort and some visible temporary upper abdominal swelling. The long-term follow-up is currently being monitored. PMID- 8415962 TI - Preoperative and postoperative nipple-areola sensibility in patients undergoing reduction mammaplasty. AB - Objective data on nipple and areola sensibility are scarce. For women with macromastia, there is little published information available indicating the incidence and intensity of postoperative nipple and areola sensibility. This prospective study was undertaken to evaluate nipple and areola sensibility in "small-breasted" control subjects as well as in patients with macromastia before and after reduction mammaplasty. Preoperative and postoperative Semmes-Weinstein pressure threshold testing was performed on 84 breasts in 43 patients and on 12 breasts of A or B cup size in the control group. The patients underwent reduction mammaplasty by the central parenchymal pedicle technique or the laterally based inferior pedicle technique. Nipple-areola sensibility was retained in 96 percent of breasts when the excision of breast tissue was less than 550 gm and 85 percent of breasts when the excision was greater than 550 gm. Overall, nipple-areola sensibility was retained in 90.5 percent of the 84 breasts tested. In those breasts in which nipple-areola sensibility was retained after surgery, there was no statistical difference in the preoperative and postoperative Semmes-Weinstein pressure threshold values. When pressure threshold values were compared in patients who had less than 550 gm of tissue resected, patients who had greater than 550 gm of tissue resected, and controls who had not undergone surgery, the trend of decreasing nipple-areola sensibility with increasing breast size was clearly seen. PMID- 8415963 TI - Treatment of extensive cranial bone defects using computer-designed hydroxyapatite ceramics and periosteal flaps. AB - We performed cranioplasty using hydroxyapatite ceramics and periosteal flaps in three patients with extensive cranial bone defects. These defects were left after post-brain surgery infection forced the removal of cranial bone. Hydroxyapatite ceramics made by using computer-aided design from three-dimensional computed tomographic image data were implanted in these patients. The defects were relatively large (the largest was 16.5 x 7.5 cm) and had a high degree of curvature. Three pieces were required in one patient, although one piece was sufficient in the other two patients. The surgical technique consisted of removal of the epidural granulation tissue, exposure of the cranial bone defect site, and shaping of the hydroxyapatite ceramics to fit the defect entirely, followed by the implantation of the hydroxyapatite ceramics. In anticipation of induction of the bone to hydroxyapatite, we covered the hydroxyapatite ceramics with periosteal flaps of cranial bones; however, based on only these three patients, our knowledge of the ossification-promoting effect is incomplete. More clinical cases should be investigated to evaluate further the clinical efficacy of this method for treatment. As we have reported here, the treatment of cranial bone defects by using computer-designed hydroxyapatite ceramics and a periosteal flap is safe and highly effective. PMID- 8415964 TI - Longitudinal assessment of mental development in infants with nonsyndromic craniosynostosis with and without cranial release and reconstruction. AB - The effect of cranial release and reconstruction on the mental development of infants with nonsyndromic craniosynostosis was evaluated. Longitudinal assessment of mental development for infants before and after cranial release and reconstruction and for infants not undergoing surgical treatment was obtained by using the mental scale of the Bayley Scales of Infant Development. Severity of anatomic craniofacial deformity, perinatal medical risk factors, and age at time of surgery also were investigated. None of the infants displayed mental retardation [Mental Development Index (MDI) score < 70] before or after cranial release and reconstruction. Scores ranged from borderline retardation to very superior following a normal distribution. Severity of anatomic craniofacial deformity and perinatal risk factors were unrelated to mental development. Cranial release and reconstruction did not affect mental development positively or negatively but did result in improvement of the original craniofacial deformity. PMID- 8415965 TI - Multidisciplinary treatment results for patients with isolated cleft palate. AB - Fifty-eight patients with cleft palate only who had received treatment in the Department of Otolaryngology-Head and Neck Surgery at the University of Iowa were examined for treatment results. Forty-one (70.7 percent) of the 58 patients showed a syndrome or suggestive factors. An unusually high percentage (36 percent) of the 58 patients required secondary surgery for velopharyngeal dysfunction or showed indications for surgery at examination. Some but not all of the relatively low success rate appears related to surgical experience. Speech proficiency, hearing acuity, and dental status were within normal limits or nearly so. The 20 patients with pharyngeal flap surgery were doing well, with minimal indications of functional obstruction. PMID- 8415966 TI - Unilateral microtia reconstruction: is the position symmetrical? AB - A method of analyzing the head position of the reconstructed unilateral microtic ear relative to the normal side was developed and applied in the assessment of 78 microtic ear reconstructions. Two groups of patients were assessed. Group I included 58 unilateral microtia patients, some with no clinical evidence of lateral facial dysplasia and others with mild to moderate lateral facial dysplasia, as judged by three experienced clinical observers. Group II included 20 unilateral microtia patients with severe lateral facial dysplasia involvement. Of special interest was evidence of lateral facial dysplasia in 88 percent of the patients reviewed. A reproducible technique was developed to assess the position of the reconstructed ear. Three slides showing the normal ear, reconstructed ear, and anteroposterior views of each patient were projected onto a centimeter grid to provide life-size (5-cm) proportions. The normal side served as the control. Length, angulation, and height were compared between normal and reconstructed ears by using a clear film rhomboid pattern designed by choosing specific reproducible landmarks. In group I, the reconstructed ear was of acceptable length and angulation in 91 and 95 percent of the patients, respectively, and of proper height in 88 percent. The corresponding values in group II were 90, 95, and 90 percent. Efforts should be made at the time of original surgery to ensure that the reconstructed ear is not only aesthetically pleasing but also positioned correctly. The technique used by us is one example of an attempt to evaluate the resultant ear position in microtia reconstruction. PMID- 8415967 TI - Management of the ear in rhytidectomy. AB - We have found that patients evaluate the results of their rhytidectomy on the basis of the frontal view. Failure to visualize the ear from the side may result in "ear neglect" on the part of the patient. The surgeon's impression of patient satisfaction may contribute to the lack of attention paid in the aesthetic literature to the ear in rhytidectomy. We present our approach for an increased awareness of the role of the ear in rhytidectomy. Our technique for analysis of the ear is based on a long-standing aesthetic standard of the ear with a well documented surgical and anatomic basis. This paper discusses how attention and consideration to the incision, axis of the ear, and size and position of the ear lobule will lead to an improved result in rhytidectomy. PMID- 8415968 TI - The submental island flap: a new donor site. Anatomy and clinical applications as a free or pedicled flap. AB - This paper describes the use of a new island flap based on the submental artery. Previously described cervical flaps have inherent problems ranging from limited mobility to unacceptable donor-site scars, to unpredictable outcomes. The flap design and technique were studied on 20 fresh cadavers and 8 patients who underwent radical neck dissections. The flap was then used successfully for reconstruction of orofacial defects in 8 patients. The flap has a long (up to 8 cm), reliable pedicle, and cutaneous dimensions can reach up to 7 x 18 cm. It can be used as a cutaneous, musculofascial (cervicofascial and platysma), or osteocutaneous flap. This flap has an excellent skin color match and a wide arc of rotation, and can extend to the whole homolateral face, except for a part of the forehead and the whole oral cavity. The anatomy, the technique, and clinical experiences are presented. PMID- 8415969 TI - Fetal tibial bone healing in utero: the effects of miniplate fixation. AB - Although clinical and experimental findings have demonstrated that fetal soft tissue wounds heal without scarring, very little is known about the process of fetal bone healing. This study examined fetal long bone healing in utero, both histologically and biochemically, with and without fracture fixation in a fetal sheep model. Our study group consisted of 25 live fetuses (from 16 ewes). There were 50 fetal tibias in this group; 12 were control, 17 were fixed (miniplate fixation), and 21 were nonfixed. A midshaft osteotomy of the tibia, either fixed or non-fixed, was performed on fetal sheep at 95 days' gestation (term = 145 days) in utero. The sheep were then killed at one of five postoperative time intervals (weeks 1, 2, 3, 4, and 7), and fetal bone healing was examined. The variables reviewed included gross morphology, histology, radiology, and collagen analysis (proportions of types II to I and III to I collagen). Fetal bone healing without fixation was accompanied by a large callus with rapid and abundant cartilage and collagen deposition. Bone healing was characterized by malunion or nonunion at 7 weeks. However, with miniplate and screw fixation, callus formation was minimal; primary bone healing occurred by 3 weeks and did not adversely affect long bone growth. Analysis of callus samples revealed a minimal amount of type III collagen, whereas the proportion of type II collagen was variable and proportional to the content of callus cartilage. PMID- 8415970 TI - A comparison of obsidian and surgical steel scalpel wound healing in rats. AB - There are several anecdotal clinical articles claiming wound healing and scar superiority using obsidian (volcanic glass) scalpels. In order to determine if skin incisions made with obsidian were superior to those made with standard surgical steel, wound tensile strength, scar width, and histology were assessed in 40 adult male Sprague-Dawley rats. Each rat received two parallel 8-cm dorsal skin incisions, one with an obsidian scalpel and the other with a surgical steel scalpel (no. 15 blade). Data were analyzed by ANOVA. Tensile strength of the two wound types was not different at 7, 14, 21, and 42 days. Scar width, however, was significantly less in the obsidian wounds at 7, 10, and 14 days (p < 0.005). At 21 days, scar width was not different in the two groups. At 42 days, all wounds were barely detectable, thus precluding scar width analysis. A blinded histologic review suggested that obsidian wounds contained fewer inflammatory cells and less granulation tissue at 7 days. PMID- 8415971 TI - Mesovascularized island flexor tendon: new concepts and techniques for flexor tendon salvage surgery. AB - Hand surgeons have long sought ways of avoiding adhesions in flexor tendon repair. We report a new concept in tendon surgery--the use of flexor tendons with perfect mesotendon blood supply and an inherent gliding mechanism composed of the paratendon and the common carpal sheath to minimize adhesions during tendon healing. This technique, inspired by reversed ulnar island forearm transfers, makes use of the flexor superficialis of the ring finger, which is supplied by a vascular mesotendon emerging from the ulnar pedicle just before Guyon's canal, and ensures a perfect blood supply, a favorable environment for suturing, and, consequently, impressive digital excursion. PMID- 8415972 TI - Microvascular reconstruction of the foot: weight-bearing patterns, gait analysis, and long-term follow-up. AB - Between 1980 and 1989, 46 free-tissue transfers (32 skin flaps, 14 muscle flaps) were performed in 44 patients for foot reconstruction. Patient age averaged 25.8 years (range 2 to 74 years). Length of follow-up averaged 43 months. Flap survival rate was 96 percent. Debulking was done in 16 skin and 8 muscle flaps. The ulceration rate in 25 patients with weight-bearing flaps was 32 versus 11 percent in 19 patients with non-weight-bearing flaps. Time before ambulation averaged 6.4 months following weight-bearing reconstruction and 4.5 months following non-weight-bearing reconstruction. All patients with either abnormal foot-mat diagrams or major gait abnormalities had significant underlying radiologic foot abnormalities or nerve injuries. We conclude that both skin and muscle free flaps frequently need secondary debulking procedures to improve function. The ulceration rate tends to be higher in weight-bearing flaps than in non-weight-bearing flaps. Underlying bony architecture and nerve function affected weight-bearing patterns and gait more than type of wound coverage. PMID- 8415973 TI - Cartilage warping: an experimental model. AB - Cadaveric cartilage was cut into blocks with a newly devised cartilage cutter. Over one-hundred pieces of cartilage were used to define a kinetics curve of cartilage warping. Kinetics curves were developed for a control group of cartilage blocks placed in saline-soaked gauze (n = 46). In addition, kinetics curves were developed for cartilage placed in hypotonic saline (n = 14), hypertonic saline (n = 14), and cyanoacrylate glue (n = 6). Photographs of all groups were taken at timed intervals in order to plot the cartilage warping. It was found that pieces of cartilage which were cut peripherally (n = 6) warped twice as much as those cut centrally (n = 40). This was significant to p = 0.001. Within 15 minutes, centrally cut pieces of cartilage warped to approximately 90 percent of their end warpage; on the other hand, peripherally cut pieces of cartilage required 30 minutes to warp 90 percent of their destined warpage. The variables used did not significantly alter the kinetics curves as compared with control. PMID- 8415974 TI - Regeneration following rejection of peripheral nerve allografts of rats on withdrawal of cyclosporine. AB - Sequential changes in the sciatic nerve function and morphology were evaluated in transplanted nerve allografts from ACI-RT1a into Lewis RT1I rats after withdrawal of cyclosporine, which had been administered subcutaneously daily (5 mg/kg) for 12 weeks. Experimental groups were established as follows: (1) allograft with cyclosporine (evaluated and sacrificed at 12, 14, 16, 20, 24, and 36 weeks, 10 rats at each week), (2) allograft without cyclosporine (12 and 24 weeks, 10 rats at each week), (3) isograft with cyclosporine (12 and 24 weeks, 6 rats at each week), and (4) isograft without cyclosporine (12 and 24 weeks, 10 rats at each week). Regeneration was evaluated through walking track analysis, electrophysiologic studies, weight of the anterior tibial muscle, and axon counts, diameter, and myelin thickness. Regeneration was observed through 14 weeks after grafting; by 16 weeks, all rats had demonstrated a clear rejection phase, but regeneration indices then recovered quickly by 24 weeks. Electron microscopy of both the graft and distal nerve suggested that both Schwann cells and axons were affected by the rejection phenomenon. Allografts without cyclosporine showed inferior regeneration histologically at 24 weeks. Morphologically, allografts were equivalent to isografts treated with cyclosporine at 24 weeks. Although nerve allografts are rejected after cyclosporine withdrawal, they may still serve as effective nerve conduits. PMID- 8415975 TI - Immunocytochemistry of skeletal muscle basal lamina grafts in nerve regeneration. AB - The influence on nerve regeneration of the extracellular matrix glycoprotein laminin was studied after sciatic nerve transection in 90 outbred Sprague-Dawley rats. Nerve regeneration through basal lamina grafts was comparable with regeneration through traditional nerve grafts across gaps up to 2.0 cm in length. True axonal regeneration rather than axonal branching was demonstrated by retrograde horseradish peroxidase labeling of nerve cables. Pretreatment of basal lamina grafts with antilaminin antibodies reduced the total number of regenerated axons by 90 percent with a significant decrease of nerve conduction velocity and a significant impairment of walking track patterns. The basement membrane glycoprotein laminin serves a critical role in the regeneration of peripheral nerves through basal lamina grafts. PMID- 8415976 TI - Take Duluth ... please. PMID- 8415977 TI - The missing postoperative photograph and other evasions. PMID- 8415978 TI - Microvascular ear replantation with no vein anastomosis. AB - In replantation of a totally amputated ear, the artery only was repaired with no vein repair. Venous stasis was successfully prevented by daily skin punctures during the first 4 days postoperatively. The elastic cartilage framework with no internally circulating blood constitutes the major percentage of the auricle mass. Thus the metabolic demand of the ear is relatively small, according to its small caliber nutrient vessels. Although the successful result in this single case report means neither a consistent procedure nor uniformly safe choice of treatment, the potential use of the single-artery repair with no accompanying vein anastomosis in ear replantations, we believe, deserves to be considered. PMID- 8415979 TI - Galactorrhea after reduction mammaplasty. AB - A case of extremely painful swelling of the breasts following a reduction mammaplasty is presented. There were no signs of an abscess or hematoma. A milky white fluid due to galactorrhea was evacuated at operation, and further galactorrhea was inhibited by medication. The pathogenesis of galactorrhea and its treatment are discussed. PMID- 8415980 TI - Use of a compression splint after immediate ear reconstruction. PMID- 8415981 TI - Standardization in reduction mammaplasty: a comparison of techniques. AB - Reduction mammaplasty is enhanced by the use of more than one technique. By comparing the results of my reduction mammaplasties, over the past 20 years, aided by computer imaging and standardization. I set up some guidelines. The following parameters were selected: age, size of breast, weight, torso build, skin tone and texture, skin color, hereditary factors, degree of ptosis, and the patient's size request. The postoperative evaluations are based on volumetric reduction of size, appearance of scar, geometric appearance of the breasts in harmony with the torso build, nipple position, and patient satisfaction. These comparisons enabled me to provide recommendations for a variety of techniques to address the needs of different patients. PMID- 8415982 TI - A template for the planning of rhombic skin flaps. AB - I have devised a template for the Limberg rhombic flap. I originally designed this template as a teaching aid, but I now find it indispensable in deciding the precise orientation of defect and flap without time-wasting and sometimes confusing trial skin marking. It is a simple, autoclavable device that can be added to any soft-tissue surgical set, and in my experience, it has significantly improved the quality of Limberg flap design. PMID- 8415983 TI - RRCPS policies on pregnancy and parental leave. Residency Review Committee for Plastic Surgery. PMID- 8415984 TI - Pregnancy and plastic surgery residency. PMID- 8415985 TI - Pregnancy and plastic surgery residency. PMID- 8415986 TI - Pregnancy and plastic surgery residency. PMID- 8415987 TI - Pregnancy and plastic surgery residency. PMID- 8415988 TI - Preparation techniques for silicone gel in the evaluation of its in vivo effects. PMID- 8415989 TI - Costochondral graft as replacement of a dysplastic mandibular condyle. PMID- 8415990 TI - Refraction changes after brow lift. PMID- 8415991 TI - "Twin-pedicled" scapular and latissimus dorsi free flaps. PMID- 8415993 TI - AIDS, aesthetic surgery, and the plastic surgeon. PMID- 8415992 TI - Hypertrophic sternal scars. PMID- 8415994 TI - Vertical mammaplasty. PMID- 8415995 TI - Conservatism in liposuction. PMID- 8415996 TI - [Investigation into the formation of proportions of "realistic thinking vs magical thinking" in paranoid schizophrenia]. AB - Both magical thinking among healthy persons and magical and symbolic thinking in schizophrenia were discussed. The investigation covered 100 paranoid schizophrenics. They also underwent an examination in connection with the formation of the remaining 3 proportions. Both "realistic thinking and magical thinking" scales were used. An ability to think realistically was preserved, to a varying degree, in all patients, with 50% of those examined having shown an explicit or very explicit ability to follow realistic thinking. The above findings deviate from a simplified cognitive model within the discussed range. It was further confirmed that realistic thinking may coexist with magical thinking, and, in some cases, it concerns the same events. That type of disorders of the content of thinking are referred to as magical-realistic interpenetration. The results, and particularly high coefficient of negative correlation within the scales of the examined proportions, confirm the correctness of the assumption that the investigated modes of thinking form an antithetic bipolarity of proportions, aggregating antithetic values, therefore being also complementary. PMID- 8415997 TI - [Research into the formation of proportions of "insight vs lack of insight" in paranoid schizophrenia]. AB - The relevant literature was reviewed, focusing on available evidence which emphasizes common occurrence of lack of insight among the sufferers of schizophrenia. The opinion has long been considered one-sided as it ignores occurrence of partial insight. Therefore, the purpose of this research was to attempt to find out to what extent the patient can afford to have insight and when he or she fails in doing so. A group of 100 paranoid schizophrenics were examined and both "insight and lack of insight" scales were applied. The results negate the hypothesis that insight does not take place in schizophrenia. A quarter of those examined displayed partial insight, with 16% of them showing pronounced insight indicating some awareness of changes in their psychic state due to "some" unidentified pathological factors. Different approaches and hypotheses were taken into consideration that would oscillate between the two extreme attitudes of insight and lack of insight. Contiguity between the examined phenomenon and that of coexistence of logically correct trains of thought with incorrect ones, as it occurs in hypomaniacal states, were indicated. An attempt to explain the so-called McEvoy's riddle was made. Final conclusion: It can be assumed that the interpenetration of the two antithetic trains of thought, where one displays partial lack of insight and the other partial insight, may be particularly characteristic of schizophrenia, more so than lack of insight. PMID- 8415998 TI - [Formation of fundamental psychopathological proportions in autistic and syntonic schizophrenias]. AB - This paper recapitulates the research conducted in recent years, including the results of the first stage of research presented in 1988. The research is characterized by meticulous preparation of 8 scales which enables us to observe the formation of 4 fundamental proportions. Of vital importance for the interpretation of the results are correlations among the proportions and among scales. Explicit correlation (r = 0.69) between the syntonic-autistic proportion and that of "vitality and emotional adequacy--emotional dementia" was recorded. This observation is of great significance in isolating paranoid schizophrenia subgroups. Emotional dementia is also significant (r = 0.64) in cases characterized by profound autism. Thus, syntony is not only a quality which strongly correlates negatively with autism (r = 0.79) but above all there exists a high positive correlation between syntony and emotional vitality (r = 0.58). Not included here is a discussion on complex relationships between syntonic autistic proportion and the one of "insight vs lack of insight". The obtained results confirm the correctness of earlier hypothesis and correlate with the results of the research on the formation of syntonic-autistic proportion. In other words, they verify a hypothesis which makes us isolate autistic and syntonic schizophrenias within paranoid schizophrenia. PMID- 8415999 TI - [Delusional psychoses during first hospitalization: age of onset and diagnosis of schizophrenia by six diagnostic criteria]. AB - The histories of illness of 167 patients with syndromological diagnosis of delusional psychosis were analysed with respect to their nosological qualification based on clinical diagnostics and 5 selected, operationalised criteria of schizophrenia. Its purpose was to establish dependence between diagnostics of schizophrenia and morbidity age (early, middle and late). A conclusion was that with application of clinical criteria, diagnosis of schizophrenia was a prevailing nosological qualification within the examined group (67%). Such diagnosis was less frequent in the group of late morbidity psychoses. It was also concluded that the dependence on morbidity age was displayed by these criteria which were strongly premised on a defined diagnostic approach. The Schneider's primary symptoms allowed more often for diagnosis of schizophrenia in later age. The Bleuler's (axial) symptoms more often facilitated diagnosis of schizophrenia in earlier age. Mediating criteria, belonging to the DSM-III-R or ICD-IO classification, did not display dependence on morbidity age. PMID- 8416000 TI - [Prognosis in delusional psychoses: comparison of prognostic value of schizophrenia by six diagnostic criteria]. AB - A group of 107 patients with diagnosis of syndromological dilusional psychosis underwent a catamnestic examination after 11.2 years from contracting the disease (8.7 years after the first hospitalization). During the first hospitalization with application of different criteria, the dependence of the early (clinical improvement after the first hospitalization) and late stages of the disease, qualified on the basis of catamnesis (intensification of the residual complex, life functioning, hospitalization intensity) on nosological qualification of disorders (schizophrenia vs no-schizophrenia) was analysed. It was concluded that the prognostisity of the course of illness depended in negligible degree on the kind of applied criteria. Of greater prognostic value, however, were the criteria in case of which diagnosis of schizophrenia depended, from the very beginning, on meeting the conditions formulated either directly (DSM-III-R) or indirectly (AS, VRC) of illness chronisity. PMID- 8416001 TI - [Brain evoked potential in schizophrenic patients]. AB - The authors reviewed the literature on evoked potentials in mentally ill patients, with particular emphasis on schizphreniacs. The commonly observed abnormalities were as follows: 1) higher SEPs amplitudes with less waveshape variability during first 100 ms in non-depressed chronic, paranoid or undifferentiated patients with florid psychotic symptoms; normal SEPs amplitudes in acute or latent schizophrenics and in chronic depressed schizophrenics but without florid psychotic symptoms; 2) reduced SEPs and VEPs amplitude recovery and faster latency recovery; 3) reduced AEPs amplitude and latency; 4) greater VEPs waveshape variability and tendency to be "reducers" in hallucinating patients; reduced amplitude and latency recovery; prolonged latencies in patients with positive family history (schizophrenia or affective disorders in close relatives); prolonged N2 latency in motor responses to "easy" and "difficult" stimuli; reduced activity of "late potentials"; 5) greater waveshape variability in all modalities in chronic schizophrenics, abnormal P300 (reduced amplitude, lack of P300 or negative "effect of uncertainty") and abnormal CNV (less "readiness" potential, prolonged negativity with motor responses). PMID- 8416002 TI - [Relationship between normal aging and Alzheimer's dementias. Practical applications]. AB - During the process of normal ageing significant loss of neural brain cells is known to take place. To a greater or lesser degree, all ageing persons undergo the process which does not deviate in a qualitative sense from the morphological changes observed in the dementias. It is therefore reasonable to assume that the following 3 relationships can occur between normal ageing and the dementias. Firstly, dementia may results directly from the ageing process of the C.N.S. alone. Secondly, a number of diverse factors either augments normal ageing deficits or accelerates the very process. Finally, normal ageing and dementias are assumed to be coincidental. There exists considerable evidence that most dementias are generated by an accelerating rate of ageing of the C.N.S. or by augmentation of the process by diverse factors. The acceptance of the above hypothesis may be beneficial not only from the theoretical point of view but could also produce new treatment techniques and, particularly, assist in preventing Alzheimer's and other dementias. PMID- 8416003 TI - [Computer tomography of the brain in psychiatric patients. Comparison with other indices of organic changes]. AB - A group of 80 psychiatric patients--40 males and 40 females--aged 13 to 72--were examined in order to compare the results of computer tomography (CT) of the brain with indicators of organic changes obtained from EEG, psychological and neurological tests. The most frequent organic pathology present in the computer tomography was brain atrophy, displayed in 40 patients (50% of the examined) and constituting the only change in 29 patients (36% of the examined, including 19 males and 10 females). The comparison of 29 patients with brain atrophy with those with normal CT revealed that the former produced pathological EEG more frequently. However, it did not indicate any predominance of pathological results of psychological and neurological examination, neither did it show one of defined psychopathological image. Within the group of "other organic pathology", the most frequent abnormality was hyperostosis frontalis interna. It was present in 6 persons, including 5 females, and was usually associated with brain atrophy and pathological results of psychopathological tests. Other abnormalities found in the "other group of organic changes" included celebral tumours, hydromas, ischenic foci and inherited defects of the brain. No explicit relationship between the CT and the indicators of organic changes was found in this group. PMID- 8416004 TI - [Organic risk factors and developmental psychopathology in a group of 100 hospitalized adolescents]. AB - In the Psychiatric Clinic for Children and Adolescents, a group of 100 hospitalized adolescents were assessed with respect to organic risk factors and developmental psychology, using original questionnaires drawn up by the clinic. Pathology during infancy, early childhood and the results of psychological tests assessing changes in the C.N.S. differentiated 4 diagnostic subgroups: psychoses, neuroses, anorexia and conduct and personality disorders. Organic risk factor indicators were most frequent in the subgroup of conduct and personality disorders. Some indicators of developmental pathology such as speech impediments, eating disorders and conduct disorders, correlated highly with psychiatric diagnosis during adolescence. PMID- 8416005 TI - Designing new psychosocial treatments for schizophrenia. AB - Schizophrenia is a disease characterized by cognitive, psychophysiological, and interpersonal deficits that result in a marked vulnerability to stress (Dawson and Neuchterlein 1984; Nuechterlein 1977; Strauss et al. 1987). Episodes of illness occur in vulnerable individuals who experience stressful life events (G. W. Brown and Rutter 1966; Lukoff et al. 1984) or stressful interactions with family members (G. W. Brown et al. 1972; Imber Mintz et al. 1987; Leff and Vaughn 1985). Similarly, overstimulating therapeutic environments have been shown to exacerbate psychosis (Drake and Sederer 1986; Liberman 1982; Linn et al. 1980; Van Putten 1976). A full understanding of disease-specific deficits resulting from stress and vulnerability is necessary for developing psychosocial treatment programs that augment pharmacotherapies in significantly ameliorating the symptoms and disabilities of schizophrenia. PMID- 8416006 TI - Studying the treatment contract in intensive psychotherapy with borderline patients. AB - A pilot study on the process of psychodynamic psychotherapy of borderline personality disorder at the Cornell University Medical College is designed to investigate the teaching and application of a specific model of treatment for borderline patients (Clarkin et al. 1992; Kernberg and Clarkin 1992). The project has involved teaching a group of self-selected trainees and faculty the manualized therapy (Kernberg et al. 1989); taping each of the twice-weekly therapy sessions over a period of 2 years; and rating (1) each therapist's adherence to the manual (Koenigsberg et al. 1985), (2) each therapist's skill, and (3) patient change. The patients are women with borderline personality disorder, between 20 and 40 years of age, diagnosed by DSM-III-R criteria (American Psychiatric Association 1987), SCID-II (Spitzer et al. 1987), and a self-report questionnaire for level of personality organization. At regular intervals, the patients are evaluated for symptom status, change in BPD criteria, and functioning. The therapists are evaluated for adherence to the manual and level of therapeutic skill. In teaching and carrying out the manualized therapy, it became clear that a critical moment in the treatment was the setting up of the treatment contract. A study was organized to look systematically at the adherence of the therapists to the model of treatment with regard to this initial phase of the therapy. PMID- 8416007 TI - The cerebral neurobiology of hope and hopelessness. AB - Hope and hopelessness are useful constructs that have been employed by clinicians in theory making regarding the pathogenesis and course of disease and in the application of various psychological and medical treatments to illness. French (1952) and Frank (1968) viewed hope as a necessary motivating force in influencing an individual to try to overcome inner psychological conflicts and seek to resolve a psychoneurosis. Melges and Bowlby (1969) classified the types of hopelessness in psychopathological processes. Perley et al. (1971), using an objective method for content analysis of small samples of speech (Gottschalk 1974), found that elevated hope scores predicted continuation of psychiatric treatment rather than dropping out. Gottschalk et al. (1967, 1969) found that hope scores derived from verbal samples predicted the duration of survival of patients with terminal cancer receiving irradiation treatment (1969) and predicted relatively favorable outcome in psychotherapy (1967). PMID- 8416008 TI - Intensive clinical studies: the psychotherapy of schizophrenia. AB - Although the value of intensive psychotherapy for persons with schizophrenia has been challenged (e.g., Gunderson et al. 1984; May 1968), there are still many clinicians and others who believe that some kind of interpersonal work with severely disordered patients, including people with schizophrenia, is extremely helpful and perhaps essential (Tomecek 1990). A crucial problem for exploring this contention is the complex interaction among four components: the measurement of efficacy, the specific nature of the interpersonal encounters, the patients' broader lives and treatment contexts, and the exact characteristics of the patients for whom these encounters may be helpful. The former blanket acceptance among many groups of the value of psychotherapy for schizophrenia is clearly not valid. But this by no means indicates that intensive interpersonal contacts of some kind for some people with schizophrenia are not important. For example, Bullard's (1960) classic paper on dealing with paranoid patients may still be extremely valuable for working with such people. In order not to bring excessively quick closure to this issue, there needs to be far more inquiry to identify specifically what kinds of contacts may be important for which people. Then the efficacy and optimal context of these contacts can be tested. PMID- 8416009 TI - Sandra: successful psychotherapeutic work with a schizophrenic woman. AB - This presentation concerns a young woman who gradually developed a schizophrenic picture on the basis of a borderline personality, and who recovered after more than 20 years of incapacitating symptoms. Today she sees her psychotherapy as decisive for the outcome. The material was collected from case records as well as from interviews with the patient and her therapist. Special emphasis is laid on the attitude and working methods of the therapist as these were viewed retrospectively by the two participants in the therapeutic process. PMID- 8416010 TI - Schizophrenic strategies of survival. AB - This quotation contains a message as to how a robotlike and dead reality comes alive, or let us say-how the innate ego potential emerges as a personal ego experience. An individual can on the basis of this felt experience leave an early isolation and enter into a personal being, that being the individual came into the world equipped to take over but has been unable to do so because of a neglected ego development with its attendant deficient capacity to integrate an inner and outer reality. PMID- 8416011 TI - Commentary: psychosocial treatment of schizophrenia. PMID- 8416012 TI - Commentary: schizophrenic patients. AB - The results of the Boston Psychotherapy of Schizophrenia Study were reported with a mixture of hopes and fears (Gunderson et al. 1984; Stanton et al. 1984). The hope was that those therapists, hospitals, and training programs that promoted and promulgated the idea that an exploratory, insight-oriented (EIO) psychotherapy was an important, usual, and practical way of treating schizophrenia would step back from this claim and see this approach in a more restricted and selective perspective. The fear was that the results would be reduced to a broader indictment of this modality such that its proper role with schizophrenic patients would be buried. Behind both this wish and this fear were the types of vivid personal experience in working with schizophrenic patients that the papers by Sandin and by Levander and Cullberg illustrate. PMID- 8416013 TI - Asceticism: creative spiritual practice or pathological pursuit? AB - Asceticism in a religious context refers to a voluntary and sustained practice of self-denial in which immediate or sensual gratifications are renounced in order to attain a higher spiritual state (Kaelber 1987). Virtually all of the major world religions have within them a way in which the individual, through ascetic practices, can strive to achieve a more thorough absorption in the sacred. Although many psychiatrists might consider any ascetic or religious practice to be pathological, others take a more neutral view by emphasizing that religious or mystical practice can also be adaptive and creative (Group for the Advancement of Psychiatry 1976). PMID- 8416014 TI - The Tridimensional Personality Questionnaire as a predictor of six-month outcome in first episode mania. AB - The authors administered the Tridimensional Personality Questionnaire (TPQ) near hospital discharge to 27 patients with first episode mania. All patients met DSM III-R criteria for bipolar disorder, manic type, as assessed by the Structured Clinical Interview for DSM-III-R. Associations of TPQ scores with operationalized outcome variables were analyzed. Outcome variables included syndromic recovery at discharge and at 6 months, syndromic recurrence, and functional recovery. Patients who failed to achieve functional recovery by 6 months had significantly higher Novelty-Seeking dimensional scores at the time of hospital discharge. This association between TPQ scores and short-term outcome suggests that elevated Novelty-Seeking scores may reflect either personality characteristics that impair functional recovery or subclinical manic symptomatology that is not reflected in other symptom measures. The TPQ may provide useful prognostic measures in patients with new onset mania. PMID- 8416015 TI - Abnormal processing of irrelevant information in schizophrenia: the role of illness subtype. AB - In a study using a trial by trial version of the Stroop color naming task, we previously found that unmedicated patients with schizophrenia show a pattern of abnormal performance characterized by increased facilitation (speeding) of color naming, color-congruent words but normal amounts of interference (slowing) of color-naming, color-incongruent words (Carter et al., 1992). Since a similar finding had recently been reported in patients with Parkinson's disease, we suggested that this finding was consistent with hypotheses about the neurobiological substrates of cognitive impairment that draw upon parallel patterns of cognitive performance in the two illnesses. We now report results from an enlarged group of unmedicated patients with schizophrenia that extend our original finding by allowing us to evaluate the role of illness subtype in abnormal performance on the Stroop task. We found that patients with the undifferentiated subtype of the disorder account for the increased Stroop facilitation effect. Patients with the paranoid subtype show their own pattern of abnormal performance, with normal amounts of facilitation and increased interference. These findings are consistent with the results of other studies which suggest that illness subtype is an important source of variability in studies of cognitive functioning in schizophrenia. PMID- 8416016 TI - Search for a genetic event in monozygotic twins discordant for schizophrenia. AB - When monozygotic twins are discordant for the diagnosis of schizophrenia, this discordance has been traditionally attributed to environmental factors acting upon a genome susceptible for the schizophrenia phenotype. The study presented here was designed to examine the occurrence of a genetic event, such as a postzygotic mitotic crossover, that could account for the discordance. Such a postzygotic event could affect cis-acting sequences and result in a phenotype of variable severity. We used molecular genetic methods to evaluate such an event with 94 microsatellite repeat polymorphic markers distributed on all autosomes and the X chromosome in five pairs of monozygotic twins discordant for schizophrenia. In this search, no genetic marker discordances were identified between the co-twins. The lack of a genetic difference may implicate nongenetic factors that are responsible in eliciting or suppressing the phenotype. However, the experiments performed in this study cannot eliminate the possibility that a tissue-specific mitotic crossover might have occurred in one of the discordant twins, which could not have been detected in our current study. PMID- 8416017 TI - A family history study of rapid-cycling bipolar disorder. AB - Previous studies have yielded mixed evidence as to whether rapid-cycling bipolar disorder (four or more episodes per year) is associated with a distinctive pattern of patient characteristics and familial aggregation of affective disorder. In this study, Family History Research Diagnostic Criteria (FH-RDC) were used to interview 165 patients with rapid-cycling bipolar disorder, non rapid-cycling bipolar disorder, or recurrent unipolar depressive disorder about the psychiatric history of 812 adult first-degree relatives. In a validity study, FH-RDC diagnoses were demonstrated to agree reasonably well with best-estimate diagnoses by two psychiatrists/psychologists, based on direct interviews with the Structured Clinical Interview for DSM-III-R. Relatives of patients with recurrent unipolar depression were less likely to have bipolar disorder and more likely to have unipolar depression than were relatives of rapid-cycling or non-rapid cycling bipolar patients. Rapid-cycling patients were younger and more likely to be female than non-rapid-cycling patients. The relatives of rapid cyclers did not differ significantly from those of non-rapid cyclers in the prevalence of bipolar disorder, unipolar disorder, rapid-cycling bipolar disorder, or substance abuse. However, there were nonsignificant trends for the relatives of rapid-cycling bipolar patients, compared with those of non-rapid-cycling bipolar patients, to have more substance abuse and less bipolar disorder given the presence of affective disorder. PMID- 8416018 TI - Obstetrical complications in patients with bipolar disorder and their siblings. AB - Although indirect evidence suggests that obstetric complications are risk factors for bipolar disorder, few studies have directly addressed this question. Probands with bipolar disorder and their adult siblings were diagnosed according to DSM III-R criteria by clinicians who had no knowledge of the subjects' obstetrical histories. Hospital records on gestations and births of 16 probands and 20 of their siblings without major mood disorders were scored for obstetric complications without knowledge of diagnosis. The assessment of obstetrical history was based on rating scales that have proved reliable and that reflect the number and severity of complications. Overall complication scores were significantly more severe in probands than siblings. Differences were most marked for perinatal complications. PMID- 8416019 TI - Growth hormone response to clonidine in neuroleptic-free patients with multidagnostically defined schizophrenia. AB - The role of alpha 2-adrenergic receptor sensitivity in schizophrenia was examined by measuring growth hormone (GH) response after the intravenous administration of clonidine (1.3 micrograms/kg) in 26 healthy control subjects and 26 neuroleptic free, acutely psychotic patients with at least 1 out of 11 possible diagnoses of schizophrenia derived from a multidiagnostic psychopathological assessment. GH responses were significantly (0.01) lower than control values in schizophrenias defined by E. Bleuler, M. Bleuler, Schneider, Langfeldt, Taylor and Abrams, and Cloninger, but not in DSM-III, World Health Organization, Feighner, Kraepelian, and Research Diagnostic Criteria (RDC) schizophrenias. Eight patients with RDC schizoaffective disorder also had a blunted response. However, there were no correlations with any symptom measures. There were no differences between paranoid and nonparanoid patients, although there was a significant difference between nonparanoid patients and control subjects. These findings support the presence of noradrenergic dysfunction in some patients within the schizophrenia syndrome, possibly those whose illnesses have an affective component. The study also illustrates the need for simultaneous investigation of several different sets of diagnostic criteria for schizophrenia in neurobiological research. PMID- 8416020 TI - Tridimensional Personality Questionnaire and serotonin in bulimia nervosa. AB - To determine the relationship between Tridimensional Personality Questionnaire (TPQ) scales and bulimia nervosa, TPQ scores of 27 bulimic women, age range 21 59, were compared with values for an age-matched sample of 128 normal control women drawn from the national norming sample by Przybeck. Scores for Novelty Seeking and Harm Avoidance were significantly higher, while scores for Reward Dependence were significantly lower for the bulimic women. A stepwise regression model of severity of purging on TPQ selected Novelty Seeking and a composite depression score, with Novelty Seeking being the stronger of the two predictors. Whole blood serotonin levels did not relate to TPQ scores or to purging frequency. PMID- 8416021 TI - Effects of intranasal vasopressin and oxytocin on physiologic responding during personal combat imagery in Vietnam veterans with posttraumatic stress disorder. AB - This study measured heart rate, skin conductance, and lateral frontalis electromyographic (EMG) responses in 43 male Vietnam veterans with posttraumatic stress disorder during personal combat imagery. In a double-blind research design, subjects were randomly assigned to receive intranasal arginine vasopressin (20 IU), placebo, or oxytocin (20 IU) an hour before the experiment. The group order of physiologic responding was as predicted: vasopressin > placebo > oxytocin. The most specific effect was exerted by vasopressin on EMG responses. This drug effect was not accounted for by nonspecific changes in responsiveness. Results are consistent with enhancing and inhibiting effects on memory retrieval and conditioned responding of vasopressin and oxytocin, respectively. PMID- 8416022 TI - Effects of the serotonin reuptake inhibitor fluvoxamine on yohimbine-induced anxiety in panic disorder. AB - To assess the effects of the selective serotonin reuptake blocker fluvoxamine on noradrenergic function in patients with panic disorder, an intravenous yohimbine challenge test was administered to eight patients with panic disorder before and after 8 weeks of fluvoxamine treatment and to a parallel group of eight patients treated with placebo. Fluvoxamine treatment reduced yohimbine-induced anxiety while placebo treatment had no effect on this variable. Both fluvoxamine and placebo treatment had little effect on biochemical or physiologic responses to yohimbine. PMID- 8416023 TI - Ubiquitination as a probe for neurodegeneration in the brain in schizophrenia: the prefrontal cortex. AB - Abnormalities in brain structure and brain function have been described in schizophrenia. It is not yet known whether these are caused by an abnormality of brain development, some form of birth injury, or a neurodegenerative process. Using immunocytochemical methods and a marker for neurodegeneration (ubiquitin), we examined an area of prefrontal cortex from elderly schizophrenic and control subjects for the presence of ubiquitin-positive degeneration products. There was no statistical difference in the degree of ubiquitination between the control and the patient samples. The findings provide no evidence to support a neurodegenerative process. PMID- 8416024 TI - Influence of partial sleep deprivation on the secretion of thyrotropin, thyroid hormones, growth hormone, prolactin, luteinizing hormone, follicle stimulating hormone, and estradiol in healthy young women. AB - The influence of partial sleep deprivation during the second half of the night on the secretion of thyroid stimulating hormone (TSH), thyroxin (T4), free T4 (fT4), triiodothyronine (T3), prolactin (PRL), growth hormone (GH), luteinizing hormone (LH), follicle stimulating hormone (FSH), and estradiol (E2) was investigated in 10 healthy young women. Blood samples were drawn at hourly intervals over a 64 hour period (i.e., 3 consecutive days and nights). During night 2, all subjects were awakened at 1:30 a.m. During partial sleep deprivation, TSH concentrations increased significantly and remained elevated throughout the following day. Levels of T4, fT4, and T3 were enhanced during the partial sleep deprivation hours only, and changes in these hormones seemed to be independent of TSH. PRL levels decreased, LH and E2 concentrations increased, and GH and FSH secretion remained unchanged during partial sleep deprivation. This pattern of change of different endocrine axes during partial sleep deprivation resembles those seen after total sleep deprivation, suggesting that similar neurochemical changes are induced by both forms of antidepressant therapy. The late evening GH peak occurred almost exclusively before the onset of sleep. Partial sleep deprivation did not influence the chronobiological profiles of any of the hormones investigated. The chemical changes underlying these alterations are speculated to involve enhancement of central norepinephrine and dopamine activity with a concomitant increase in the activity of the sympathetic nervous system. PMID- 8416025 TI - Serum homovanillic acid levels in schizophrenic patients and normal control subjects. AB - Schizophrenic patients with an early age at onset of illness had low baseline levels of homovanillic acid (HVA) in serum compared with schizophrenic patients with a late age at onset. After adjustments were made for age at onset, there was a significant partial correlation between positive symptoms and serum HVA. The relationship between positive symptom scores and serum HVA was shifted to the left in the early onset patients, suggesting a relatively increased sensitivity of dopamine-associated response. Patients with severe negative symptoms also had an earlier age at onset and a trend toward lower serum HVA. This study found no difference between mean serum HVA values in schizophrenic patients and normal control subjects. PMID- 8416026 TI - Aggression and the endangered self. AB - Discussions of aggression in the psychoanalytic literature are dominated, both explicitly and implicitly, by loyalty to, or rejection of, Freud's theory of the aggressive drive, which has caused an unfortunate polarization. An approach to aggression is presented that preserves essential features of both drive theory and nondrive theory traditions. It is suggested that this way of thinking about aggression is actually prevalent in both contemporary Freudian and non-Freudian thought without being recognized as such. PMID- 8416027 TI - Sexualization and desexualization. AB - This paper is a reconsideration of the role of sexualization in psychoanalysis. After a review of the accepted definitions of sexualization as defense and/or part of normal sexuality, a new explanation is offered. This posits sexualization as a psychological manifestation of a structural deficit and suggests analytic treatment as a process of structuralization. These terms are defined and illustrated. A case example demonstrates the thesis, and some implications of this concept are discussed. PMID- 8416028 TI - Self and sign in free association. AB - Some depressive symptoms may be mitigated early in psychoanalytic treatment, apparently as the consequence of the free association process itself. We may account for these changes by considering possible modification of the self as the container of hitherto unavailable signifying chains, now expressed in the analytic discourse. At the same time, it is useful to consider how similar favorable changes may arise in other forms of treatment, and also in experiences of ordinary life. PMID- 8416029 TI - The perverse attitude toward reality. AB - Freud's (1940a) tentative distinction between the defensive maneuvers in neurosis and those in perversion can be extended to good clinical effect. In general, neurotic defenses may be thought of as directed against wishes, whereas perverse defenses are directed against perceived reality. It is suggested that the perverse approach to reality is not limited to frank sexual perversions; it defines a class of operations that involve taking certain liberties with reality. Clinical material is used to demonstrate the perverse attitude and some of its implications for technique. The role of the superego is considered. PMID- 8416030 TI - The word "just": an essay on resistance, words, and multiple meanings. AB - This is an essay on the word, just, often used by patients as an ordinary trivial word but also as part of resistance functioning to derail further associations. But there are other, more weighty meanings of the word connected with justice and fairness, and thus, ethics and morality. Besides these, there are hidden meanings that certain words can convey consciously and unconsciously. "Just" is part of a "family" of words, some now obsolete, that range from aspects of law to various passionate derivatives of sexuality and aggression. Therefore, the word, just, can serve as an example to which the insights of psychoanalysis can be applied when seeking answers to questions about the origins, hidden meanings, changes, and apparent losses of meaning of other "ordinary" words. PMID- 8416031 TI - Do left-handers die sooner than right-handers? Commentary on Coren and Halpern's (1991) "Left-handedness: a marker for decreased survival fitness". AB - Cross-sectional life span studies of handedness typically show decreasing percentages of left-handers in older age groups. In an article in Psychological Bulletin, S. Coren and D. F. Halpern (1991) argued that this age trend reflects the shorter life span of left-handers than right-handers. They presented 2 studies of their own providing what they regard to be direct evidence that left handers, on average, die sooner than right-handers. They also proposed a variety of reasons for what they called left-handers' "decreased survival fitness." I discuss Coren and Halpern's reasons for rejecting a more conventional explanation of the life span data; the 2 studies that they offered in support of their argument; their analysis of other evidence they invoked to account for left handers' putative decreased survival fitness; and, finally, new studies in which the longevity explanation was tested by more direct means than have been used thus far. I conclude that the case for the "decreased survival fitness" hypothesis cannot be sustained. PMID- 8416032 TI - Mediators and moderators of change in cognitive therapy of depression. AB - This article reviews the theoretical and empirical literature associated with the mediators and moderators of change in cognitive therapy (CT) of depression. Covariation between change in cognition and change in depression, specific effects for cognitive versus behavioral components of CT, specific effects for CT versus other treatments, moderating influence of nonspecific and technical aspects of the therapeutic environment, and moderating influence of client characteristics are reviewed. The clinical implications of prior research and suggestions for future research on identifying the critical ingredients of change in CT of depression are discussed. PMID- 8416033 TI - Predictors of differences between Type A and B individuals in heart rate and blood pressure reactivity. AB - Past estimates of the magnitude of Type A-B differences in cardiovascular reactivity are probably overly conservative. In addition, it is unclear which situations are more likely to elicit excessive reactivity in Type As. The present meta-analysis found that, overall, Type As had greater heart rate (mean d = .22), diastolic blood pressure (d = .22), and especially systolic blood pressure responses (d = .33) than Type Bs; these effect sizes were small but relatively consistent. However, Type As showed especially greater cardiovascular reactivity in situations characterized as having (a) positive or negative feedback evaluation, (b) socially aversive elements such as verbal harassment or criticism, and (c) elements inherent in playing video games. Measures of time urgency, Type A assessment method, and gender were not found to be strongly related to A-B differences in cardiovascular reactivity. Future studies that use more "Type A-relevant" situations will probably find greater effects. PMID- 8416034 TI - Cardiac psychophysiology and autonomic space in humans: empirical perspectives and conceptual implications. AB - Contemporary findings reveal that autonomic control of dually innervated visceral organs does not lie along a single continuum extending from parasympathetic to sympathetic dominance. Rather, a bivariate autonomic space bounded by sympathetic and parasympathetic axes is the minimal representation necessary to capture the modes of autonomic control. We here empirically instantiate a quantitative bivariate model for the chronotropic control of the heart in humans. This model provides a more comprehensive characterization of psychophysiological response than simple measures of end-organ state and permits a differentiation of behavioral states and processes that would otherwise remain obscure. The model also illuminates and subsumes general principles such as the law of initial values and reveals a fundamental physiological rationale for the selection of heart period over heart rate as a metric for cardiac chronotropy. The present article also considers strategies for psychophysiological investigations within the autonomic space model, the limitations of these methods, and analytical tools for assessing their validity. PMID- 8416035 TI - Taxonomy, assessment, and diagnosis of depression during adolescence. AB - Research on depressive phenomena during adolescence has focused on 3 separate constructs: depressed mood, depressive syndromes, and depressive disorders. Approaches to the assessment, taxonomy, and diagnosis of these 3 conceptualizations are reviewed. Each of the approaches is represented by different assessment tools measuring related but distinct aspects of depressive phenomena. The constructs share a common set of symptoms reflecting negative affectivity but differ in their inclusion of symptoms of anxiety, somatic problems, and disrupted concentration and in the duration and severity of the symptoms they include. Depressed mood, syndromes, and disorders are integrated as 3 levels of depressive phenomena in a hierarchical and sequential model, and moderating factors are hypothesized to account for the relationships among the 3 levels. The need for a stronger developmental focus to understand depressive phenomena during adolescence is emphasized. PMID- 8416036 TI - Mathematical disabilities: cognitive, neuropsychological, and genetic components. AB - Cognitive, neuropsychological, and genetic correlates of mathematical achievement and mathematical disability (MD) are reviewed in an attempt to identify the core deficits underlying MD. Three types of distinct cognitive, neuropsychological, or cognitive and neuropsychological deficits associated with MD are identified. The first deficit is manifested by difficulties in the representation or retrieval of arithmetic facts from semantic memory. The second type of deficit is manifested by problems in the execution of arithmetical procedures. The third type involves problems in the visuospatial representation of numerical information. Potential cognitive, neuropsychological, and genetic factors contributing to these deficits, and the relationship between MD and reading disabilities, are discussed. Finally, suggestions for the subtyping of mathematical disorders are offered. PMID- 8416037 TI - Statistical difficulties of detecting interactions and moderator effects. AB - Although interaction effects are frequently found in experimental studies, field researchers report considerable difficulty in finding theorized moderator effects. Previous discussions of this discrepancy have considered responsible factors including differences in measurement error and use of nonlinear scales. In this article we demonstrate that the differential efficiency of experimental and field tests of interactions is also attributable to the differential residual variances of such interactions once the component main effects have been partialed out. We derive an expression for this residual variance in terms of the joint distribution of the component variables and explore how properties of the distribution affect the efficiency of tests of moderator effects. We show that tests of interactions in field studies will often have less than 20% of the efficiency of optimal experimental tests, and we discuss implications for the design of field studies. PMID- 8416038 TI - Metrical information load of lines and angles in line patterns. AB - Acute angles and long lines are more complex than obtuse angles and short lines. This source of complexity is called metrical information, for which a measure was proposed by Leeuwenberg (1982). According to this measure, the metrical information of a static pattern is specified as the sum of the mechanical impulses necessary to move an object according to this pattern. The perceptual relevance of this metrical load was tested in an experiment in which judged complexities were gathered for a number of simple line patterns. PMID- 8416039 TI - A diffusion model of early visual search: theoretical analysis and experimental results. AB - Meinecke (1989, Exp. 1, cond. HO) showed that the detectability of a visual target embedded in a linear noise array decreases with increasing retinal eccentricity, while the reaction time (RT) of the hits increases. One of the most interesting features of her results was that the RT of the correct rejections is consistently larger than the RT for signals presented near the fovea. This finding suggests that initially visual attention is concentrated near the fixation point and then diffuses across the stimulus array to perform a serial, exhaustive search. We present a diffusion model of early visual-search processes that quantitatively describes this evolution of attention in time and space; in contrast to most previous conceptions, it is based on a genuine relation between the spatial and temporal dimensions of the search processes performed. The model predicts quantitatively both detection performance and RT. We conducted an experiment similar to that of Meinecke (1989), but with an additional variation of the presentation time. All the main features of the 189 predictions could be explained by the model. The interpretation of the four model's parameters is discussed in some detail and compared with previous estimates of the microscopic search speed derived from alternative models. Finally, we consider some possible modifications related to results of Kehrer (1987, 1989), and some generalizations to multi target detection and two-dimensional stimulus arrays. PMID- 8416040 TI - The role of attention for the Simon effect. AB - It has been claimed that spatial attention plays a decisive role in the effect of irrelevant spatial stimulus-response correspondence (i.e., the Simon effect), especially the way the attentional focus is moved onto the stimulus (lateral shifting rather than zooming). This attentional-movement hypothesis is contrasted with a referential-coding hypothesis, according to which spatial stimulus coding depends on the availability of frames or objects of reference rather than on certain attentional movements. In six experiments, reference objects were made available to aid spatial coding, which either appeared simultaneously with the stimulus (Experiments 1-3), or were continuously visible (Experiments 4-6). In contrast to previous experiments and to the attentional predictions, the Simon effect occurred even though the stimuli were precued by large frames surrounding both possible stimulus positions (Experiment 1), even when the reference object's salience was markedly reduced (Experiment 2), or when the precueing frames were made more informative (Experiment 3). Furthermore, it was found that the Simon effect is not reduced by spatial correspondence between an uninformative spatial precue and the stimulus (Experiment 4), and it does not depend on the location of spatial precues appearing to the left or right of both possible stimulus locations (Experiment 5). This was true even when the precue was made task relevant in order to ensure attentional focusing (Experiment 6). In sum, it is shown that the Simon effect does not depend on the kind of attentional operation presumably performed to focus onto the stimulus. It is argued that the available data are consistent with a coding approach to the Simon effect which, however, needs to be developed to be more precise as to the conditions for spatial stimulus coding. PMID- 8416041 TI - The effects of size, clutter, and complexity on vanishing-point distances in visual imagery. AB - The portrayal of vanishing-point distances in visual imagery was examined in six experiments. In all experiments, subjects formed visual images of squares, and the squares were to be oriented orthogonally to subjects' line of sight. The squares differed in their level of surface complexity, and were either undivided, divided into 4 equally sized smaller squares, or divided into 16 equally sized smaller squares. Squares also differed in stated referent size, and ranged from 3 in. to 128 ft along each side. After subjects had formed an image of a specified square, they transformed their image so that the square was portrayed to move away from them. Eventually, the imaged square was portrayed to be so far away that if it were any further away, it could not be identified. Subjects estimated the distance to the square that was portrayed in their image at that time, the vanishing-point distance, and the relationship between stated referent size and imaged vanishing-point distance was best described by a power function with an exponent less than 1. In general, there were trends for exponents (slopes on log axes) to increase slightly and for multiplicative constants (y intercepts on log axes) to decrease as surface complexity increased. No differences in exponents or in multiplicative constants were found when the vanishing-point was approached from either subthreshold or suprathreshold directions. When clutter in the form of additional imaged objects located to either side of the primary imaged object was added to the image, the exponent of the vanishing-point function increased slightly and the multiplicative constant decreased. The success of a power function (and the failure of the size-distance invariance hypothesis) in describing the vanishing-point distance function calls into question the notions (a) that a constant grain size exists in the imaginal visual field at a given location and (b) that grain size specifies a lower limit in the storage of information in visual images. PMID- 8416042 TI - Codes and operations in picture matching. AB - Ellis and Allport (1986; see also Ellis, Allport, Humphreys & Collis, 1989) proposed a model of object perception wherein successively more abstract descriptions are generated as a function of processing time. The aspect of their model that is examined here is the proposal that viewer-centred representations of objects decay rapidly whereas object-centred or semantic-level representations do not. To test the model, a picture-matching task was used in which subjects decided whether successively presented pictures rotated in the frontal plane had the same name. The pictures were either identical pictures, pictures of different objects with the same name, or pictures of objects with different names. The two successive pictures could be in the same orientation or in a different orientation. In Experiment 1, two orientations (0 degrees upright and 120 degrees) and two ISIs were examined (100 ms and 2 s). In Experiment 2, two orientation (0 degrees and 60 degrees) and three ISIs were examined (100 ms, 2 s, and 5 s). In neither experiment was there any evidence that viewpoint-specific representations disappeared at longer ISIs. These results, although consistent with other research on the perception of rotated objects, did not replicate the results of Ellis and Allport (1986) and are inconsistent with their model. PMID- 8416043 TI - Perceiving, its component stream of perceptual experience, and Gibson's ecological approach. AB - Gibson's theory nearly explicitly distinguishes the activity or process of perceiving from its component stream of perceptual experience (awareness). An activity of perceiving is a total process of a perceiver's using a perceptual system to perceive something in the environment or of himself or herself in that environment. An activity of perceiving includes, inter alia, an obtained stimulus energy flux at the respective receptors, as well as a stream of perceptual experience (awareness) which proceeds at certain brain centers of the respective perceptual system. Obtaining stimulation, though this be highly structured and nomically specific to environmental properties, is not the having of perceptual experience (awareness); in addition to information pick-up, there must take place, in the nervous system, extraction of informational features (invariants and variants) of the stimulus energy flux. But the Gibsonian Lombardo argues that perceptual awareness is not a brain process; it occurs, rather, at the ecological level of organization. In effect, this contradicts Gibson's theory, which holds (a) that information pick-up, but not extraction, occurs at the interface between perceiving and environment, and (b) perceptual experience (awareness), in contrast to perceiving, is not publicly observable, as it would be by definition if it occurred at the ecological level of organization. PMID- 8416044 TI - Prolactin response to dl-fenfluramine in panic disorder. AB - The objective of this study was to test the hypothesis that serotonin receptors are hypersensitive in patients with panic disorder. Eleven patients and 12 controls received a single PO dose of 60 mg of dl-fenfluramine at 0900h on a single occasion. Blood samples were collected with an indwelling intravenous catheter at 30-min intervals from 0930h to 1530h and prolactin determined by radioimmunoassay. In both groups, fenfluramine induced a rise in the plasma prolactin concentration from baseline. The patients showed a greater increase in prolactin response than the normal controls. This result is consistent with the hypothesis of increased serotonin receptor function in patients with panic disorder. PMID- 8416045 TI - Personality characteristics and platelet MAO activity in women with congenital adrenal hyperplasia (CAH). AB - Personality traits and platelet monoamine oxidase (MAO) activity were studied in 22 women, 17-34 years old, with prenatal virilization due to congenital adrenal hyperplasia (CAH) (21-hydroxylase deficiency) and 22 healthy controls. The CAH group differed significantly on two of the eight scales of the Karolinska Scales of Personality (KSP), which have earlier shown significant gender differences. Both differences were in the masculine direction, with a high, male level, score for Detachment and a lower score for Indirect Aggression. The Detachment scale reflects distance in social relations, and has earlier been shown to be strongly gender differentiating. There was no significant difference in platelet MAO activity between the CAH group and the controls. Although an influence of psychosocial factors cannot be excluded, the results suggest a possible association between prenatal androgen exposure and the high Detachment score for the CAH group. Gender differences in empathy, affiliation motivation, intimacy and maternal behavior may be relevant parallels. PMID- 8416046 TI - Cortisol concentrations in serum of borderline hypertensive men exposed to a novel experimental setting. AB - We tested the effect of repeated exposure to a novel experimental setting on resting, morning blood cortisol concentrations in borderline hypertensive men. Borderline hypertension is thought to be accompanied by increased central nervous system activation, which would predict enhanced anticipation and cortisol secretion in a novel experimental situation. Twenty-two borderline hypertensive and 23 normotensive male volunteers were tested in four sessions all separated by 2 or more days. Borderline hypertensives had higher cortisol concentrations than normotensives during the first and second days (p < .005) but not on the third and fourth days. Significant reductions in cortisol concentrations across days were noticed in borderline hypertensives but not in normotensives. All subjects reported feeling more activated on the first day than on any other day (p < .01). These results indicate that exposure to a novel experimental situation may enhance secretion of stress related hormones such as cortisol in borderline hypertensives. PMID- 8416047 TI - Dependency of growth hormone (GH) stimulation following releasing hormones on the spontaneous 24-hour GH secretion in healthy male and female subjects. AB - The purpose of this investigation was to evaluate whether in healthy subjects the GH response following stimulation with releasing hormones is dependent on the spontaneous GH secretion within 24 hr prior to the stimulation test. In 18 male and 9 female healthy subjects (21-59 years) GH was measured every 15 min over 26 hr. Twenty-four hours after the beginning of blood sampling, a GH stimulation test was performed by using a combined releasing hormone test. Sleep was recorded in three consecutive nights. A positive correlation was found between the AUCs of the 24-hr GH secretion and the AUCs of GH stimulation, which could not be explained by an age effect only. This study demonstrates that subjects with comparatively high amounts of GH secreted within 24 hr also show good GH secretory responses when immediately after the 24-hr sampling period a stimulation test is undertaken. Therefore, a low GH response to stimulation cannot be explained by feedback effects of high GH amounts secreted during the 24 hr before the test or by empty pituitary GH storages. PMID- 8416048 TI - Consequences of restraint stress on natural killer cell activity, behavior, and hormone levels in rhesus macaques (Macaca mulatta). AB - Three experiments were performed to determine the effect of stress on the neuroendocrine-immune system in nonhuman primates. In Experiment 1 the diurnal variation in cell and hormone levels was determined. The percentages of neutrophils, monocytes, and eosinophils fluctuated throughout the 24-hr period, while white blood cell (WBC), neutrophil-to-lymphocyte ratio (N:L), hemoglobin (Hgb), natural killer cell cytotoxicity (NK activity) and beta-endorphin levels did not. Experiment 2 investigated the effects of ketamine and restraint on behavior. Scratching was increased in control monkeys and animals receiving ketamine, whereas passivity was increased in chair-restrained animals. In Experiment 3, eight adult male rhesus monkeys were restrained in primate chairs at 0600h. Behavior was filmed for 3 hr and blood samples were collected at 0700, 0800, and 0900. Whole blood was analyzed for total WBC and percentage of each leukocyte type. NK activity was also measured. Plasma levels of cortisol and beta endorphin were determined and behavior was quantitated from video-records. WBC and the percentage of neutrophils increased during the restraint period, while the percent lymphocytes and monocytes decreased. NK activity also decreased over time after restraint whereas plasma cortisol and beta-endorphin levels increased significantly. Although after the 3 hr of restraint stress, changes were found in hormone levels, behavior, and NK activity, there were no significant correlations between the parameters measured. Thus, our results indicate that there is not a common neuroendocrine response or single neuroendocrine mediator that results in predictable behavioral changes and immune suppression following stress. PMID- 8416049 TI - Stressful life events and affective disorders inhibit pulsatile LH secretion in hypothalamic amenorrhea. AB - The aim of this study was to evaluate relationships between emotional state and hypothalamic activity in patients with hypothalamic secondary amenorrhea. Sixty seven normal weight patients with hypothalamic amenorrhea were submitted to concomitant psychological and LH pulsatility evaluation. Structured clinical interview for anxiety and depressive disorders (DSM III-R) as well as life events investigations (Paykel test) were performed. LH pulses (blood sampling every 10 min for 4 hr) were analyzed through DETECT program and Instantaneous Secretory Rate were computed. Twenty-one patients reporting life events associated to the onset of amenorrhea had LH pulse frequency (2.28 +/- 1.10 pulses/4 hr) lower than those without life events (3.40 +/- 1.46 p = .007). LH pulses amplitude was lower in patients meeting a DSM III-R (21 cases: 1.22 +/- 0.96 mIU/ml) diagnosis than in those without (1.99 +/- 1.20 p = .04) diagnosis. Plasma estradiol and FSH levels as well as duration of amenorrhea and Body Mass Index were similar among groups. It is concluded that psychogenic factors (namely the presence of life events related to the onset of menstrual disorder) are associated with significant and specific changes of hypothalamic activity which could be involved in determining hypogonadism. PMID- 8416050 TI - Effects of beta-funaltrexamine treatment and sexual isolation in the perinatal period on the development of mu-opioid receptors and nociception. AB - Male and female rats were segregated from birth (sexually isolated animals). Additional litters containing both male and female pups were kept as controls (mixed-housed animals). beta-FNA (5 mg/kg) or water was administered SC at day 0 or day 7 to isolated and mixed-housed animals. The development of mu-opioid receptors and nociceptive responses in the two groups was assessed at day 7 or 14, respectively. Mu-receptor binding was measured in whole brain using (3H) DAGO as a binding ligand and nociception assessed using the tail immersion test. beta FNA treatment depressed mu-receptors when measured 1 but no 7 days later. However, male and female rats treated at day 0 with beta-FNA had lower brain protein content. Sexual isolation had little effect on mu-receptor number and did not augment the beta-FNA effect. However, isolation increased pain sensitivity in 7-day-old animals and in 14-day-old females. beta-FNA treatment had little effect on nociceptive threshold but reversed the effects of sexual isolation. PMID- 8416051 TI - No evidence for menstrual synchrony in lesbian couples. AB - Menstrual synchrony was investigated in a sample of 29 cohabiting lesbian couples, ranging in age from 22 to 48 years. One or both partners kept prospective daily records of variables including menses onset dates, intimate contact, and sexual activity. All women reported daily intimate interaction with their partners; none reported intimate interaction with men. Despite these potentially optimal conditions for the manifestation of synchrony, the differences between dyad members in menses onset dates were distributed randomly, and there was no evidence of convergence. In fact, most dyads exhibited divergence of onset dates. Reasons for lack of synchrony in this sample are discussed; one conclusion is that there is no solid evidence that menstrual synchrony is a stable attribute of past or contemporary human populations. PMID- 8416052 TI - Thyroid function before and after four-week light treatment in winter depressives and controls. AB - Thyroid function in patients in a current major depressive episode during the course of recurrent major mood disorder with seasonal pattern according to DSM IIIR was compared to that of controls before and after 4 weeks' light treatment, and to that of controls at baseline and after 4 weeks' of arising early without exposure to bright light. No consistent abnormalities in thyroxine, free thyroxine index, triiodothyronine, reverse triiodothyronine, thyrotropin, thyrotropin response to TRH infusion, or thyroid autoantibodies were seen in depressives at baseline. No differences in these parameters were seen at baseline between depressives and controls. No intergroup differences were seen with treatment, although reverse T3 decreased significantly during the protocol in all groups. These data do not support the hypothesis that the thyroid axis plays a role in the pathogenesis of winter depressive symptoms or their response to light treatment. PMID- 8416053 TI - Short-lasting behavioural effects of thyrotropin-releasing hormone in depressed women: results of placebo-controlled study. AB - The rapid and short-lasting behavioral effects of thyrotropin-releasing hormone (TRH) were investigated in female patients with DSM-III-R major depression syndrome (MDS). Twenty-six depressed patients free of any medication received 0.2 mg of Protirelin (synthetic TRH) intravenously and 16 received placebo. All patients completed the Zung Self-Rating Depression Scale and the Spielberger State and Trait Anxiety Inventory (SSAI and STAI) twice: before and 2 hr after protirelin or placebo administration. The significant improvement in patients' emotional state after TRH injection was observed on STAI (p < .001) and SSAI (p < .01). Protireline was superior to placebo on STAI (p < .005). There was no significant correlation between behavioral effects of Protirelin and changes in thyroid hormones and TSH secretion. The improvement in patients' emotional state was more evident in depressed patients without associated panic attacks than in MDS with panic. These findings suggest that TRH has rapid positive effects on depression and that they depend more on patients' emotional state than on the function of the hypothalamus-pituitary-thyroid axis. PMID- 8416054 TI - Effects of thyroid hormones on central nervous system in aging. PMID- 8416055 TI - Effects of aging on event-related brain potentials and reaction times in an auditory oddball task. AB - Auditory event-related potentials (ERPs) were recorded from 71 healthy individuals between 18 and 82 years of age during performance of a disjunctive reaction time task in an auditory oddball paradigm. The effects of aging on reaction times and on the latencies, amplitudes, and distributions of each of the main ERP components were examined. No significant slowing of the reaction times of the elderly subjects was observed in relation to the younger ones. The peak latencies of both the N1 and P2 components elicited by standard tones were slightly but significantly slowed with age. In the ERPs of target tones, the later, endogenous components (N2, P3, and SW) showed linear increases in latency as a function of age; the later the component, the longer the age-related delay. In general, aging was associated with less negativity (both N2 and SW) and more positivity (P3) over the anterior scalp, together with a smaller P3 and a more pronounced N2 over posterior scalp areas. Most of the effects observed in target ERPs were also evident in the difference waves derived from subtraction of the standard from the target ERPs, although the slope of the age-related latency increase of N2 was shallower and that of the P3 was steeper in the difference ERPs. These findings are discussed in relation to previous accounts of ERP changes with aging. PMID- 8416056 TI - Validation of surface EMG as a measure of intravaginal and intra-abdominal activity: implications for biofeedback-assisted Kegel exercises. AB - This study validates surface EMG as a measure of pelvic muscle and abdominal activity by showing its high correlation to internal pressure data. Using standardized scores, between-subjects correlation of perineal EMG and intravaginal pressure was r = .75, and the correlation of abdominal EMG and intra abdominal pressure was r = .72. Discriminant validity was also demonstrated by showing low correlation between standardized abdominal and perineal EMG measurements (r = .10). A repeated measures multivariate analysis of variance demonstrated that visual and auditory biofeedback of EMG during pelvic floor contractions increases intravaginal pressure when compared with trials without biofeedback. Potential benefits of fabric electrodes include reduced invasiveness and risk and the ease with which patients can utilize this technology for home practice. PMID- 8416057 TI - Regional cerebral blood flow during experimental phobic fear. AB - Positron emission tomographic measurements of regional cerebral blood flow (rCBF) were used to investigate central nervous system correlates of fear and anxiety. Volunteers with symptomatic snake phobia were studied while exposed to visual phobogenic, aversive, and neutral stimuli. Anxiety ratings and the number of nonspecific electrodermal fluctuations increased as a function of phobic stimulation. Phobic, compared to neutral and aversive, stimulation elevated rCBF in the visual associative cortex. The basal ganglia were not activated more by phobic than aversive or neutral stimulation. However, cortical and thalamic rCBF were always correlated during phobic but not aversive or neutral stimulation. This indicates that the thalamus could be a relay station for phobic stimulus processing and affect. PMID- 8416058 TI - Event-related brain potential correlates of self-reported hunger and satiety. AB - Perception of hunger and satiety was examined by means of event-related potential (ERP) recordings. On the basis of Helson's frame-of-reference theory, it was hypothesized that hunger is perceived with reference to an actual adaptation level. ERPs were recorded to adjectives (hungry, thirsty, tired) combined with one of five adverbs (very, almost, somewhat, hardly, not). Subjects indicated whether a particular adverb-adjective combination accurately described their actual feeling of hunger, thirst, or fatigue. They were tested after they had fasted for 16 hr and another time when they were satiated. Area of P300 of the ERP was smallest for adverb-adjective combinations agreeing with the subject's actual state of hunger (i.e., adaptation level), which was almost hungry in fasting subjects and hardly hungry in satiated subjects. These P300 minima were flanked by significantly enhanced (p < 0.001) P300 areas for the immediately adjacent adverbs combined with hungry. P300 areas for adverbs combined with thirsty and tired did not depend on the subject's state of hunger. The data suggest that perception of internal states, such as hunger, refers to an adaptation level that is sensitively indicated by P300 area. PMID- 8416059 TI - The association between lithium carbonate and smooth pursuit eye tracking among first-episode patients with psychotic affective disorders. AB - The association between treatment with lithium carbonate and smooth pursuit eye tracking performance was investigated in first-episode patients with psychotic affective disorders. The horizontal pursuit performance of patients with major depression and bipolar disorder who were receiving lithium carbonate was contrasted with that of patients not receiving lithium carbonate. In addition, the accuracy and quality of pursuit eye tracking was examined in bipolar patients whose lithium status changed from the time of initial testing to the time of retest 10 months later. For the combined group of depressed and bipolar patients, treatment with lithium carbonate was not associated with worse pursuit performance. Bipolar disordered patients on lithium did not differ in tracking proficiency from those not on lithium; bipolar patients whose lithium status changed from intake to retest also did not display a significant change in pursuit performance. PMID- 8416060 TI - Predicting ambulatory blood pressure during school: effectiveness of social and nonsocial reactivity tasks in black and white adolescents. AB - We evaluated a newly developed stress task, the Social Competence Interview, and three nonsocial tasks (video game, mirror drawing, mental arithmetic) for ability to predict ambulatory blood pressure in 237 black and white adolescents. Blood pressure was measured in laboratory, classroom, and transition (between-class) settings. A resting laboratory baseline explained 10-49% of the variance in ambulatory blood pressure levels; the ability of the stress tasks to explain additional variance was assessed in multiple regression analyses. Only the blood pressure response to the interview enhanced prediction of classroom and transition systolic and diastolic pressures in the total sample and in blacks, whites, females, and males--even when the interview data were entered into a hierarchical regression model after those for the other three tasks were entered. Mirror drawing improved prediction of transition systolic blood pressure in the total sample, and mental arithmetic plus the interview improved prediction of classroom diastolic pressure in black males; however, video game failed to enter any predictive equation. Racial subgroup analyses disclosed that the interview data predicted systolic pressure in whites but predicted diastolic pressure in blacks, indicating biological differences in blood pressure regulation. An interview that elicits characteristic thoughts and social behaviors appears to represent a promising approach to examining environmental influences on blood pressure. PMID- 8416061 TI - Patterns and stability of cardiovascular responses to variations of the cold pressor test. AB - Test-retest reliabilities and patterns of heart rate and blood pressure responses were examined using variations in the cold pressor test in 113 normotensive white college men. Comparisons were made of stimulus site (forehead vs. foot) and bodily posture (seated vs. supine) across four separate groups of men. The stability of cardiovascular responses was examined over a 2-week-test-retest interval. Different cardiovascular response patterns emerged as a function of stimulation site and posture. Systolic and diastolic blood pressure increases were accompanied by bradycardia in the forehead cold pressor task but by tachycardia in the foot cold pressor task. Systolic blood pressure increases were larger for foot than for forehead stimulation. Heart rate increases were larger for supine than for seated men. Effects on response were independent of postural differences at baseline, and there were no stimulation site by posture interactions. The cardiovascular responses to stimulation did not attenuate across sessions in any experimental condition but were more reliable for foot than for forehead stimulation and for supine than for seated posture. Short-term stability for changes to the task approached that for baseline and task and was higher than has been reported elsewhere. PMID- 8416062 TI - Semantic priming and stimulus degradation: implications for the role of the N400 in language processing. AB - Two experiments explored the effects of stimulus degradation on behavioral and event-related potential (ERP) measures of semantic priming. The primary goal was to help elucidate the psychological processes that underlie the N400 component. In both experiments, subjects made speeded lexical decisions to words and pseudowords preceded by either semantically related or unrelated prime words. In one block of trials, the target stimuli were intact, and in a second block they were degraded by removing a random 33% of the elements making up each letter of the target (Experiment 1) or by overlaying a matrix of dots on the target (Experiment 2). In both experiments, subjects responded faster and more accurately to related targets than to unrelated targets (behavioral semantic priming effect), and this priming effect was greater when the target was degraded. However, although the N400 component was larger for unrelated than related targets (ERP semantic priming effect), there was no evidence that this difference was larger in the degraded block of either experiment. These results indicate that the behavioral and ERP measures reported here appear to be tapping into different components of the process(es) involved in semantic priming. The implication of the results for the linguistic processes underlying the N400 are discussed. PMID- 8416063 TI - Correspondence between brain ERP and behavioral asymmetries in a dichotic complex tone test. AB - Electrophysiologic correlates of perceptual asymmetry for dichotic pitch discrimination were investigated in 20 normal subjects. Brain event-related potentials (ERPs) elicited by dichotic pairs and binaural probe tones in the Complex Tone Test (Sidtis, 1981) were recorded from homologous scalp locations over left and right hemispheres (F3, F4; C3, C4; P3, P4; O1, O2). Baseline-to peak amplitudes were measured for N100, P200, and a late positive complex consisting of P350, P550, and slow wave. A left ear advantage (LEA) was evident in 70% of the subjects, and hemispheric asymmetries related to this behavioral asymmetry were found for P350 and P550 amplitudes to probe stimuli. Subjects with a strong LEA had greater amplitudes over the right hemisphere than the left, whereas subjects with little or no LEA showed a nonsignificant trend toward the opposite hemispheric asymmetry. Hemispheric asymmetry of these late ERPs at parietal and occipital sites was highly correlated with behavioral asymmetry. These findings suggest the utility of electrophysiological measures in assessing hemispheric asymmetries for processing complex pitch information. PMID- 8416064 TI - Interstimulus interval and the selective-attention effect on auditory ERPs: "N1 enhancement" versus processing negativity. AB - The attention effect on the auditory event-related potential (ERP) in dichotic conditions was studied as a function of the interstimulus interval (ISI). Subjects attended to stimuli delivered to a designated ear and responded to infrequent pitch deviants in this input. The mean ISI was either 80, 160, 480, or 800 ms. Negative difference waves (Nds) were computed by subtracting ERPs to unattended standards from ERPs to the same stimuli when attended. The exogenous N1, as estimated from unattended standard ERPs, was larger contralaterally to the stimulus and inverted in polarity at mastoids. With decreasing ISIs, N1 diminished in amplitude much faster than did the Nd. In addition, N1 latency remained stable, whereas Nd peaked markedly earlier with shorter ISIs, almost perfectly coinciding with the exogenous N1. However, this temporal coincidence found in grand averages proved to be illusory in single subjects. The early Nd showed no contralateral asymmetry at its peak, but asymmetry was apparent during the ascending slope. These lateral asymmetries resembled those of the exogenous N1 but occurred later. The early Nd peak was, at least mainly, caused by an endogenous attention effect, the processing negativity (PN), even with very short ISIs, but an effect on the exogenous N1 could not be excluded. PMID- 8416065 TI - Frontal brain asymmetry and emotional reactivity: a biological substrate of affective style. AB - Individuals differ dramatically in the quality and intensity of their response to affectively evocative stimuli. On the basis of prior theory and research, we hypothesized that these individual differences are related to variation in activation of the left and right frontal brain regions. We recorded baseline brain electrical activity from subjects on two occasions 3 weeks apart. Immediately following the second recording, subjects were exposed to brief positive and negative emotional film clips. For subjects whose frontal asymmetry was stable across the 3-week period, greater left frontal activation was associated with reports of more intense positive affect in response to the positive films, whereas greater right frontal activation was associated with more intense reports of negative affect in response to the negative film clips. The methodological and theoretical implications of these data are discussed. PMID- 8416066 TI - On the neural generators of the P300 component of the event-related potential. AB - The triarchic model of P300 amplitude (Johnson, 1986, 1988a) postulated that the overall amplitude of the P300 recorded at any given electrode site represented the summation of activity from different neural generators, each related to the processing of a different type of information. However, neither of these original accounts provided an explicit description of the methods required to establish experimentally the presence of multiple neural sources. This paper reviews the triarchic amplitude model, the subsequently obtained data that support the postulated presence of multiple generators underlying the P300, and the methods used to demonstrate the presence of these multiple sources. These methods are straightforward because it is only necessary to show that the portions of P300 amplitude associated with different experimental variables have different scalp distributions. The implications of the multiple-generator basis of P300 on such factors as component definition, neural source analyses, and the cognitive processes underlying its activity are discussed. PMID- 8416067 TI - Distortion of ERP averages due to overlap from temporally adjacent ERPs: analysis and correction. AB - In studies of event-related potentials (ERPs), short interstimulus intervals (ISIs) are often employed to investigate certain neural or psychological phenomena. At short ISIs, however, the ERP responses to successive stimuli may overlap, thereby distorting the ERP averages. This paper describes a signal processing approach for analyzing the distortion of ERP averages due to such overlap. In general, the distortion is modeled in terms of mathematical convolutions of the ERP waveform elicited by each type of adjacent stimulus with the corresponding distribution in time of those stimuli relative to the averaging epoch. Using this framework, a number of implications of ERP overlap for experimental design and interpretation are examined, with special emphasis given to selective attention paradigms. It is shown that the possibility of confound due to ERP overlap is widespread in short-ISI experiments, and even the widely used procedure of stimulus randomization does not necessarily control for differential distortion of the ERPs to attended versus unattended stimuli. Problems due to ERP overlap can be particularly serious in short-ISI studies that examine how ERPs (and associated perceptual processes) are influenced by the nature of the preceding stimulus (i.e., stimulus sequence effects). A set of algorithms is presented for estimating and removing the residual distortion due to response overlap from recorded ERP averages. The use of these algorithms, collectively termed the Adjacent Response (Adjar) Technique, can alleviate many of the overlap-related problems that arise when short ISIs are used, thereby enhancing the power of the ERP technique. PMID- 8416068 TI - An evaluation of the automaticity of sensory processing using event-related potentials and brain-stem reflexes. AB - Selective attention effects on reflexes and evoked potentials are reviewed with the aim of evaluating three theories regarding sensory automaticity. (a) The peripheral-gating theory, which assumes that ignored stimuli can be filtered out soon after transduction, was tentatively rejected because neither auditory-nerve nor retinal potentials are reliably affected by attention. (b) At the other extreme, the assumption that sensory analyses are obligatory and cannot benefit from attentional resources (i.e., strong-automaticity theory) was also rejected, because longer latency components were found to be modifiable by attention. (c) An intermediate theory provides the best fit to present electrophysiological data. The earliest sensory analyses are assumed to be strongly automatic and then, at forebrain levels, there is a transition from strong to weak automaticity (i.e., analyses are obligatory but modifiable by attention). This transition can begin as early as about 15 ms for audition and about 80 ms for vision. PMID- 8416069 TI - Effects of intermodality change on electrodermal orienting and on the allocation of processing resources. AB - Two experiments studied the effects of intermodality change on electrodermal responses and on reaction time to a secondary task probe stimulus after 24 habituation training trials with either a tone or a vibrotactile stimulus. The probe was a visual stimulus of 500 ms duration, and within-stimulus probes occurred 300 ms following stimulus onset. Experiment 1 crossed change versus no change with modality of the training stimulus. Skin conductance responses (SCRs) were larger in the experimental group than in the control on the test trial, and in the experimental group, test trial responses were larger than those on the first training trial. Probe reaction time was slower on the test trial in the experimental group than in the control, and within-stimulus probe reaction time was slower than interstimulus interval reaction time early in the habituation series. Experiment 2 crossed change versus no change with the presence of a secondary task. Test trial SCRs were larger in the experimental group than in the control, regardless of whether or not the secondary task was present. In addition, test trial responses in the experimental group were larger than those on the first training trial in both the task and no-task conditions. Within the task condition, reaction time was slower in the experimental condition than in the control on the test trial. In addition, reaction time in the experimental condition was slower during the change trial than during the first training stimulus. The data provide difficulties for noncomparator theories of habituation and seem to be most easily explained by theories of habituation that emphasize the importance of an extrapolatory process. PMID- 8416070 TI - Attention and mismatch negativity. AB - The mismatch negativity (MMN) component of the auditory event-related potential (ERP) is elicited by infrequent, physically deviant stimuli in a sequence of frequent homogeneous stimuli (standards). It has been suggested that the MMN is generated by an automatic (attention-independent) neural mismatch process with a memory trace that encodes the physical features of the standard stimulus. The proposed MMN independence of attention was addressed in the present study. Standard stimuli and two types of deviant stimuli, differing from standards either in frequency or intensity, were dichotically presented in random order and at a rapid rate. The subject attended either to left- or right-ear stimuli, counting the number of a designated type of deviants in that ear. In the present conditions of very strongly focused attention, the MMN was elicited even by frequency change in the ignored input stream, and its amplitude was very similar to that of the MMN elicited by equivalent deviant stimuli (targets) in the attended input stream. In contrast, the MMN to intensity deviation was clearly attenuated in the absence of attention. This effect is, however, probably due to the attention effect on the MMN generator itself rather than the antecedent sensory-analysis and -storing functions. PMID- 8416071 TI - The reliability of ERP components in the auditory oddball paradigm. AB - Nineteen adolescents (average age 15 years, 3 months) were tested and retested using a standard 40 target, auditory oddball ERP paradigm across an interval of 1 year, 10 months to determine reliability of the ERP components, both in terms of intersubject stability and score agreement and in terms of trait (between-session reliability) versus state (within-session reliability). Significant trait stability was found for the N100, P200, and P300 latencies (r = .48, .51, and .74, respectively), and for P300 amplitude (r = .62), supporting the P300 as a reliable measure, with the stability required for group research but not necessarily for clinical applications. Discussion and examples illustrate the application of reliability information to the planning and evaluation of ERP paradigms. PMID- 8416072 TI - Backward masking and skin conductance responses after conditioning to nonfeared but fear-relevant stimuli in fearful subjects. AB - The present study examined two issues. Are skin conductance responses conditioned to fear-relevant stimuli, as contrasted with responses conditioned to fear irrelevant stimuli, elicited after merely an automatic, nonconscious analysis of the stimulus content? Do fearful subjects show better conditioning to nonfeared but fear-relevant stimuli (e.g., conditioning to spiders in snake-fearing subjects) than do nonfearful subjects? Subjects afraid of snakes, but not of spiders, or vice versa (n = 32) and nonfearful subjects (n = 32) were shown either fear-relevant stimuli (snakes or spiders and rats) or fear-irrelevant stimuli (flowers and mushrooms) in a differential conditioning paradigm, where one of the stimuli was followed by an electric shock. During a subsequent extinction phase, the conditioned stimuli were presented under backward masking conditions, preventing their conscious recognition. Consistent with our hypothesis, during the masked extinction of the conditioned stimuli, differential skin conductance responses to conditioning and control stimuli remained only for subjects conditioned to fear-relevant stimuli. Both fearful and nonfearful control subjects had significantly larger differential electrodermal responses to fear-relevant than to fear-irrelevant stimuli. However, contrary to our hypothesis, fearful subjects did not show enhanced conditionability to their nonfeared but fear-relevant stimuli as compared with nonfearful control subjects. PMID- 8416073 TI - On the relation between cardiodynamics and heartbeat perception. AB - The mediation of heartbeat perception by cardiodynamic parameters was investigated by experimentally introducing changes in heart performance. Twenty five subjects (14 women, 11 men) underwent bicycle ergometric exercises (0-75 W) at different tilt angles (90 degrees-0 degree). After each of the 31 tilt/exercise phases, the following measures of cardiac performance were taken using impedance cardiography: stroke volume, Heather Index (as an indicator of contractile force), heart rate, and an estimate of the momentum of the ejected blood mass. Additionally, a heartbeat perception task was performed. The different tilt/exercise combinations induced profound changes in all cardiodynamic measures. The correlation between heartbeat perception and cardiodynamic parameters was highest for "momentum" (r = .67) and for stroke volume (r = .59). From our data, we infer that intraindividually occurring changes in cardiac perception are mediated partially by characteristics of heart dynamics and that the source of these interoceptive signals lies in the heart or in its close vicinity. PMID- 8416074 TI - Early and late VEPs for reading stimuli are altered by common binocular misalignments. AB - Effects of mild binocular anomalies on early and late evoked response complexes to word stimuli were examined in visual evoked potentials (VEPs) recorded over occipital cortex, Wernicke's area, and its right side homolog in university students with (fixation disparity group) and without (normal group) fixation disparity. Stimuli were monocularly or binocularly viewed words of a paragraph presented individually for 100 ms, one per second, and a binocular control condition without linguistic content. An early complex (P125-N170) recorded at Oz and a late complex (N170-late P) recorded from the temporoparietal placements were analyzed. The early complex was not influenced by linguistic content. Binocular stimuli generally resulted in larger early and late VEP amplitudes than did monocular stimuli except that the fixation disparity group had no binocular amplitude enhancement at Oz. Left hemisphere amplitude was greater than right for all conditions, more so for words than for nonlinguistic stimuli. For words, the left hemisphere advantage was significant only for the normal group. Relationships between basic visual processing and language processing in relation to early and late complexes are considered from several perspectives. PMID- 8416075 TI - Respiratory sinus arrhythmia, cardiac vagal tone, and respiration: within- and between-individual relations. AB - Respiratory sinus arrhythmia (RSA) is frequently employed as an intra- and interindividual index of cardiac parasympathetic tone, although the relationship of RSA to interindividual differences in cardiac vagal tone remains questionable. Our study examined between- and within-subject relations among RSA, cardiac vagal tone, and respiratory parameters. Twenty-nine young adults performed two sessions of tasks under no medication and single and double autonomic blockade (intravenously administered propranolol and atropine). Parasympathetic tone was determined from heart period responses to complete vagal blockade. Results indicated the following. Resting RSA does not accurately predict individual differences in cardiac vagal tone. However, RSA and heart period together do predict such individual differences reasonably well. The relationship between individual variations in RSA and vagal tone is not improved by controlling respiratory parameters. Substantial cardiac vagal activity occurs during inspiration, and intraindividual variations in respiratory measures confound the association between RSA and cardiac vagal tone. PMID- 8416076 TI - An event-related potential study of semantic congruity and repetition in a sentence-reading task: effects of context change. AB - This experiment examined the joint effects of semantic congruity and repetition on event-related brain potentials (ERPs) elicited in a sentence priming task. In an initial training phase, subjects were familiarized with a list of 60 congruous and incongruous sentences. During the second phase, ERPs were recorded as subjects silently read a set of 180 unconnected sentences. One third of the sentences were presented exactly as they had been seen in training (Old sentences), one third were presented for the first time (Completely New sentences), and one third involved a re-pairing of the frames and completions of the congruous and incongruous Old sentences (New Pair sentences). The N400 congruity effect was reduced for Old as compared with Completely New and New Pair sentence completions. These results suggest that N400 reflects processes that are sensitive to both existing semantic associations and representations of previous episodes that include the context of the eliciting stimulus. A late positive component (LPC) involving a sustained positive shift in the waveform after 600 ms was largest for incongruous completions and occurred somewhat earlier for Old sentences. This pattern of results is consistent with the notion that the LPC is an index of episodic retrieval and elaborative processes. The data also suggest the presence of an early onset slow positive shift that is only evident for New Pair congruous sentences. PMID- 8416077 TI - Stimulus presentation rate dissociates sequential effects in event-related potentials and reaction times. AB - The present study explored the impact of the stimulus presentation rate on sequential effects in event-related potentials (ERPs) and reaction times (RTs). Random series of equiprobable tones were presented at interstimulus intervals (ISIs) of 1.3, 2.1, and 2.9 s. Fast and accurate choice responses to the tones were required. Although sequential effects in RTs were stable across all ISIs, the common first-order repetition effect in P300 amplitude was only observed at the 1.3-s ISI and not at the slower presentation rates. This dissociation between the first-order effects in RTs and ERPs speaks against an explanation of both effects by a common expectancy mechanism. In addition, sequential effects were observed as early as about 100 ms after stimulus onset in the lateralized readiness potential. Together with similar sequential effects in P300 latency, this finding supports a continuous flow model of information processing. PMID- 8416078 TI - An alternative method for significance testing of waveform difference potentials. AB - Guthrie and Buchwald (1991) proposed an ad hoc procedure for assessing the statistical significance of waveform difference potentials that may arise in a variety of psychophysiology research contexts. In our paper, an alternative method is presented and demonstrated that has fewer underlying assumptions than does the Guthrie-Buchwald test and may, therefore, produce better results in some situations. In particular, the test proposed here (a) is distribution free, (b) requires no assumption of an underlying correlation structure (e.g., first-order autoregressive), (c) requires no estimate of the population autocorrelation coefficient, (d) is exact, (e) produces p values for any number of subjects and time points, and (f) is highly intuitive as well as theoretically justifiable. This procedure may be used to carry out multiple comparisons with exact specification of experiment-wise error, however, this test is based on permutation principles and may require large amounts of computer time for its implementation. PMID- 8416079 TI - A simple test of the Law of Initial Values. AB - The conceptualization and testing of the Law of Initial Values remains a controversial issue. We critically review common procedures to test the Law, and a simple test of the Law is described and illustrated. The reduction of variance from the pretest to the posttest is crucial in this test. PMID- 8416080 TI - Does activation of the baroreceptors reinforce differential Pavlovian conditioning of heart rate responses? AB - High- and low-pitched tones (CS+ and CS-) signalled baroreceptor stimulation or inhibition (US+ and US-) on 6-s conditioning trials (n = 128). Baroreceptor stimulation was induced by the phase-related external suction (PRES) method of Rau et al. (1992) in which a brief pulse of negative external pressure is applied to the neck at systole and one of positive pressure at diastole within each cardiac cycle (the reverse sequence is used for baroreceptor inhibition). Changes in heart period (R-R intervals) confirmed that PRES manipulated the baroreceptors in the presence of CS+ and CS- without habituation over conditioning trials. However, conditioned heart period responses were not observed on test trials (n = 32) in which CS+ and CS- were presented with the baroreceptor manipulation removed. Subjects were unable to state which CS had signalled baroreceptor stimulation and inhibition when given PRES-alone trials after the conditioning phase (differential attention thus controlled). These results (a) confirm that the differential effect of the two PRES stimuli was specific to the baroreceptors and (b) support earlier studies that have found that differential conditioning is impaired when CS-US relations are not processed in attention. We discuss implications regarding when baroreceptor firing might be discriminable and reinforcing. PMID- 8416081 TI - Tonotopic auditory cortex and the magnetoencephalographic (MEG) equivalent of the mismatch negativity. AB - Two tone stimuli, one frequent (standard) and the other infrequent (a slightly higher, deviant tone), were presented in random order and at short intervals to subjects reading texts they had selected. In different blocks, standards were either 250, 1,000, or 4,000 Hz, with the deviants always being 10% higher in frequency than the standards of the same blocks. Magnetic responses elicited by the standard and deviant tones included N1m, the magnetoencephalographic equivalent of the electrical N1 (its supratemporal component). In addition, deviant stimuli elicited MMNm, the magnetic equivalent of the electrical mismatch negativity, MMN. The equivalent dipole sources of the two responses were located in supratemporal auditory cortex, with the MMNm source being anterior to that of N1m. The dipole orientations of both sources in teh sagittal plane depended on stimulus frequency, suggesting that the responses are generated by tonotopically organized neuronal populations. The tonotopy reflected by the frequency dependence of the MMNm source might be that of the neural trace system underlying frequency representation of auditory stimuli in sensory memory. PMID- 8416082 TI - Pictures as prepulse: attention and emotion in startle modification. AB - The effects of an emotional stimulus prepulse on probe startle response were examined here. Pleasant, neutral and unpleasant pictures were viewed for 6 s, and an acoustic startle probe was presented either 300, 800, 1,300, or 3,800 ms after slide onset, or 300 or 3,800 ms after slide offset. Blink magnitude and onset latency demonstrated (a) an early (prepulse) inhibition effect in which reflexes elicited immediately after slide onset were smaller than reflexes elicited later in the viewing interval, and (b) affective modulation, in which unpleasant stimuli prompted larger reflexes than pleasant. Interactive effects of probe time and picture valence indicated attention/arousal effects early and pleasantness effects late in the picture interval. Effects of both attention and emotion can be simultaneously measured using this startle-probe paradigm, encouraging its use in both basic and clinical contexts. PMID- 8416083 TI - Definition of a responder: analysis of behavioral, cardiovascular, and endocrine responses to varied workload in air traffic controllers. AB - Individual differences in behavioral, cardiovascular, and endocrine responses to varying workload among 381 air traffic controllers were assessed using random regression modeling. Although most men showed significant increases in behavioral arousal associated with increasing planes, there were major individual differences in response in systolic and diastolic blood pressure, heart rate, and cortisol. Approximately 20% to 25% of those studied had large increases in each of these domains, along with a smaller group showing inverse responses in heart rate and cortisol. There was also evidence of a smaller number of enhanced responders within the highest groups, who tended to have more missing values at higher levels of workload. There was convergence in the definition of responders using three statistical strategies: random regression, correlational analyses, and ANOVA. Response in one physiological/behavioral domain was essentially independent of response in another, supporting the conclusion of specificity, rather than a global tendency to respond to increasing work load. PMID- 8416084 TI - Abnormal stress responses in patients with diseases affecting the sympathetic nervous system. AB - Diseases that cause malfunction of the sympathetic nervous system provide insight into how the sympathetic nerves normally modulate responses to stress. This paper discusses insight from a number of such diseases. Transection of the cervical spinal cord leads to autonomic dysreflexia. This syndrome causes episodic hypertension in quadriplegic patients from excess sympathetic activity reflexly activated by bowel or bladder distention. These patients lack cerebral control of spinal sympathetic reflexes. Radiotherapy to the neck can destroy the arterial baroreceptors that monitor blood pressure fluctuations. Patients who lack baroreceptors have exaggerated blood pressure responses to stress. They have episodes of hypertension and hypotension that cause headaches and dizziness. Diabetics and uremics often develop a peripheral sympathetic neuropathy. They have postural hypotension and diminished blood pressure responses to stress. They are often unable to tolerate heat, exercise, or fluid deprivation. Patients with heart failure deplete sympathetic neuronal norepinephrine stores. The continual stress of heart failure diminishes their ability to respond to further stresses such as standing upright or exercising. Patients with diseases of the sympathetic nervous system illustrate that everyday occurrences such as a change in posture or ambient temperature are stresses requiring a marked change in sympathetic nervous activity. Both physical and psychological stresses elicit large initial sympathetic neuronal responses that are subsequently damped by feedback inhibition from structures such as the baroreceptors. Damage to part of these feedback loops leads to exaggerated pressor responses to stress. PMID- 8416085 TI - Regular exercise and aerobic fitness in relation to psychological make-up and physiological stress reactivity. AB - This study assessed the association of aerobic fitness with psychological make-up and physiological stress-reactivity in a group of untrained men, as well as the effects of 4 and 8 months of exercise training on these parameters. Psychological assessment included questionnaires on personality (Neuroticism, Type A, Hostility), coping styles (Anger In, Anger Out), negative affect (Depression, Anxiety), and self-esteem. Stress reactivity was measured as the cardiovascular and urinary catecholamine response to two competitive reaction time tasks and the cold pressor test. No cross sectional relationships were found between aerobic fitness, defined as the maximal oxygen consumption during an exhaustive exercise test, and any of the psychological variables. In addition, psychological make-up did not change as a consequence of exercise training. In further contrast to our hypothesis, aerobic fitness was associated with high, rather than low, cardiovascular reactivity. Longitudinal effects of training were limited to a reduction in the overall levels of heart rate and diastolic blood pressure. This suggests that regular exercise does not increase the resistance to stress-related disease by influencing psychological make-up or acute psychophysiologic reactivity. PMID- 8416086 TI - Stress and immunity in humans: a meta-analytic review. AB - This article presents a meta-analysis of the literature on stress and immunity in humans. The primary analyses include all relevant studies irrespective of the measure or manipulation of stress. The results of these analyses show substantial evidence for a relation between stress and decreases in functional immune measures (proliferative response to mitogens and natural killer cell activity). Stress is also related to numbers and percent of circulating white blood cells, immunoglobulin levels, and antibody titers to herpesviruses. Subsequent analyses suggest that objective stressful events are related to larger immune changes than subjective self-reports of stress, that immune response varies with stressor duration, and that interpersonal events are related to different immune outcomes than nonsocial events. We discuss the way neuroendocrine mechanisms and health practices might explain immune alteration following stress, and outline issues that need to be investigated in this area. PMID- 8416087 TI - The concept of neurosis: a reassessment is overdue. PMID- 8416088 TI - Cognitive interventions in behavioral medicine. AB - In this report an overview is given of the contribution of cognitive approaches to behavioral medicine. The (possible) contribution of cognitive therapy is reviewed in the area of coronary heart disease, obesity, bulimia nervosa, chronic pain, benign headache, cancer, acquired immunodeficiency syndrome/human immunodeficiency virus and asthma. Although the relative contribution of cognitive therapy varies across these various disorders, its positive effects are now well established and new advances undoubtedly will be made in the next few years. PMID- 8416089 TI - Psychosocial factors in gastrointestinal illness. AB - Gastrointestinal (GI) illnesses represent a paradigm of psychosomatic medicine. Nearly half of patients seen in GI practice present with functional illnesses, and patients commonly complain of symptoms that have coexisting organic and functional etiologies. This chapter addresses the connected nature of psychosocial factors and GI function, disease and outcome in the context of the biopsychosocial model, which allows illness to be examined from the encompassing perspective of interacting system, from the cellular to the environmental. This perspective also helps explain why biologic events such as oncogene alteration can produce heterogeneous clinical and biological responses. Links between gut and brain, involving neuroendocrine associations of the enteric nervous system and its connections with the spinal, autonomic and central nervous systems, are well documented. Neural connections allow information to affect GI secretion and motility. Disturbances in one component of the system can lead to brain-gut effects, such as dysmotility and mood disturbance. Appropriate diagnosis and treatment require clear understanding of biologic, psychologic, and social contributory events. In chronic unexplained GI illnesses, the appropriate clinical approach may be to suspend the search for structural disease, and work instead to assess and treat the physical symptoms and psychosocial problems at hand. PMID- 8416090 TI - In-patient and ward psychosomatic psychotherapy: concepts, effectiveness and curative factors. AB - Based on the situation in the Federal Republic of Germany, this article reviews the development, organization and theoretical framework of in-patient and ward psychosomatic psychotherapy. Indications for treating psychosomatic patients in hospital, the effects of treatment and some results on the process of in-patient psychotherapy are summarized. Some of the specific features of in-patient and ward psychotherapy are adopted as components for a theoretical model of in patient psychotherapy. Our experience with this form of therapy suggests that it can play an important role within the care system, is a fruitful field for training psychotherapists and acts as an excellent model for further research into psychotherapy. PMID- 8416091 TI - Content analysis of follow-up interviews with inpatient/outpatient group psychotherapy patients. AB - In the follow-up project of the psychosomatic clinic at the University of Heidelberg, 45 patients were interviewed who had begun group psychotherapy in a closed group over 2.5 years with an inpatient treatment period of 3 months. The follow-up interviews carried out 2-5 years after the end of the outpatient treatment period are studied by content analysis. It could be demonstrated that the content of the two main categories 'Beginning' and 'End' of the inpatient period differs very much. Two patterns of 'beginners' could be distinguished. The importance of the evaluation of the former patients' spontaneous report as involved experts for psychotherapy research is discussed. PMID- 8416092 TI - Is there a typical conflict in gastric complaints? An epidemiologic psychoanalytic contribution to the question of conflict specificity in psychosomatic medicine. AB - The question of conflict specificity for subjects with gastric complaints was investigated in the context of a representative, epidemiologic-psychoanalytic field study. Out of a population of 600 adults probands with dyspepsia were compared with individuals with musculoskeletal symptoms and 'healthy' subjects. Using interview protocols 2 'blinded' raters judged whether a dependency independency conflict was present in different aspects of life. Findings supported the higher prevalence of the conflict among subjects with dyspepsia. Therefore, the conflict was considered typical, but not specific for the experimental group. PMID- 8416093 TI - Alexithymia and movie preferences. AB - The goal of this study was to further our understanding of the relationship between the ability to identify one's emotions and the kinds of emotion arousing experiences that people prefer. One hundred and eighty-six undergraduate students completed the Toronto Alexithymia Scale, the Beck Depression Inventory, and a questionnaire inquiring about subjects' movie preferences. Individuals who reported having difficulty identifying their emotions were more likely than those without such difficulty to prefer negatively valenced movies relative to happy movies. Individuals who had both high levels of depression and alexithymia were more likely to prefer fear-arousing rather than anger-arousing movies. PMID- 8416094 TI - An investigation of hostile and alexithymic characteristics in breast cancer patients. AB - Two measures of personality characteristics, the Personality Deviance Scale and the Toronto Alexithymia Scale, were administered to a total sample of 448 women and a subsample of 100 women, respectively, attending two breast-screening centers in Athens. The relative risk of breast cancer between women with low denigratory attitudes towards others and women with high scores in this scale was 2.31. Statistically significant relations were not noted between the mean scores of alexithymia factors and breast cancer diagnosis. Our findings point to the conclusion that breast cancer patients can be distinguished from healthy women on the basis of their cognitive activity, characterized by rare denigratory thoughts. PMID- 8416095 TI - Cortisone image and emotional support by key figures in patients with bronchial asthma. An empirical study. AB - In a subgroup of patients with bronchial asthma, irrational fears of steroid medication can be observed beyond justified worries about side-effects. A cortisone image which involves overemphasizing the threatening aspects of cortisone often underlies non-compliant illness behaviour. In the present study, cortisone image and the relationship with a key figure was investigated in 62 patients with bronchial asthma, all of whom had participated in a structured asthma treatment and teaching programme (ATTP). The data show that (1) perceiving cortisone as threatening predicts poor adherence to ATTP guidelines how to manage severe attacks, and (2) patients having a supportive relationship with their key figure are less likely to experience threatening aspects of cortisone. PMID- 8416096 TI - Alexithymia and the recognition of facial expressions of emotion. AB - Slides of photographs depicting posed facial expressions of nine different emotions were presented to 131 females and 85 males who were asked to identify the emotion(s) being experienced by the person in each photograph. Subjects were then administered the 20-item version of the Toronto Alexithymia Scale; the 33rd and 66th percentiles were used to categorize subjects into high, moderate, and low alexithymia groups. Results showed that the high alexithymia group was significantly less able to recognize facial expressions of emotions than the low alexithymia group. There was no significant effect for gender on the ability to recognize facial emotions. The results suggest the presence of deficits in the perception of nonverbal emotion in alexithymia. PMID- 8416097 TI - The impact of dream interpretation using psychological kinesiology on the frequency of recurring dreams. AB - Subjects reporting a recurring dream at least 4 times a month for a year or more were assigned to a dream recording control group and an experimental group which used muscle testing to guide dream interpretation. Dream frequency was recorded prior to and following a dream interpretation intervention (experimental subjects) or dream reporting session (control subjects). Dream frequency declined in the experimental group, suggesting that recurring dream frequency is a useful dependent variable, and that psychological kinesiology dream interpretation is a powerful intervention. PMID- 8416098 TI - Robert Kellner, MD, PhD. PMID- 8416099 TI - The neurobiology of stereotypies. PMID- 8416100 TI - [Coping with occupational stresses in health professions]. AB - A model of coping with job stressors is presented, whereby the mediating effect of coping always is goal directed. There exists a variety of stressors in health professions which call for flexible adaptation. Results of studies by this author on coping with job stressors in nursing and dentists are presented. Coping strategies of nurses are not always optimal: in factor analysis coping by will and problem solving was followed by social support; less favourable is diversion and compensation by consumption, fatalistic-depressed withdrawal or aggressive acting out. Religions support seems to be especially helpful in a subgroup. Job satisfaction remains high (80% satisfied), with progressive satisfaction going with little stress values, but poor satisfaction with high stress. In the coping process of doctors and dentists there is a better correspondence between stressors and coping strategies. Quite in contrast to the rich literature on stressors there are few studies on coping of healthcare professionals, so that it is difficult to derive general conclusions. Nevertheless, based on qualitative analysis (repertory grid by Kelly) certain typical patterns were elaborated like "manager; denier; pleaser; problem seeker". Finally intervention strategies to improve coping of health professionals are reviewed. PMID- 8416101 TI - [Polyarthritis patients with and without detected rheumatoid factor: a comparison of the psychological personality]. AB - The study examined again the question whether the presence of rheumatoid factor in rheumatoid arthritis (rh.A.) patients has a bearing on their personality scores. For this purpose 20 seropositive rh.A. patients were compared by means of the FPI-R (Freiburg Personality Inventory) with 20 rh.A. patients in whom the rheumatoid factor was absent. Groups were matched for sex and age; severity and duration of disease were taken into account as covariates. The covariates showed significant correlations with some personality variables. Diagnostic subgroups, however, did not differ in any of the FPI scales. PMID- 8416102 TI - [Interplay between defenses and coping during the course of illness--a study of Crohn disease patients]. AB - The psychological coping concept and the psychoanalytic theory of defense are the two most important models for assessing coping with chronic illness. Their interplay has hardly been investigated so far. A "layer model" is proposed to describe the connections between defense and coping. This model is empirically tested by applying a path analysis (latent trait model) to patients suffering from Crohn's disease during the course of the illness. The layer model can be verified, but is valid only during the acute phase of the illness. The results convincingly show that the relationship between defense and coping can only be described as a dynamic interplay that changes in the course of the disease process. PMID- 8416103 TI - [The therapeutic relationship within the scope of inpatient psychotherapy]. AB - Within psychotherapy-research a kind of paradigm has almost established itself, maintaining that--with high probability--a positive therapeutic relationship early on permits to expect a favourable therapeutic outcome. These conclusions were mainly derived from an outpatient psychotherapy setting of under 30 hours. In our own study we wanted to examine whether an early prognosis of success under such conditions was also possible for psychoanalytically oriented inpatient psychotherapy. We examined a sample--representative by age, sex and diagnosis--of 76 patients with an average duration of treatment of 12 weeks. For the patients' self-rating of therapeutic alliance we translated a questionnaire from Luborsky (1984) into German, whereas for the therapists' rating we conceived our own questionnaire. Contrary to our expectations, we were not able to demonstrate clinically significant correlations between the initial quality of the therapeutic relationship and therapy success later on (patients' rating: r = 0.28, p < 0.01, therapists' rating: r = 0.09, p > 0.10). One hypothesis on this surprising finding seems to be especially plausible: In our opinion a substantial strength of inpatient psychotherapy (regardless of it being shaped more by psychoanalytic or behavioral orientation) is the following: it offers less motivated patients of lower social status and level of education an opportunity to experience essential processes of communicative learning and motivation especially within the first weeks of treatment. PMID- 8416104 TI - [Spontaneous awakening and dreams of patients with psychophysiologic sleep disorders]. AB - The effect of dreaming on the formation of psycho-physiological sleep disorders is studied by an investigation of 26 patients and 15 control persons who were interviewed as to the occurrence of dream memory after spontaneous awaking and after being awaken systematically in the sleep laboratory. It turned out that the patients had less dream memory than the control persons after spontaneous awaking from REM sleep, but they did not differ after being awaken. In addition, dream reports after spontaneous awaking from REM sleep contained more abstract dream thoughts and less visible dream action. These findings are interpreted as a manifestation of disturbed "dream work" in patients with psychophysiological sleep disorders, i.e. disturbance of connecting thoughts and scenes from the unconscious. Thus, awaking in sleep disorder is interpreted to be a progression into awakeness as a matter of defense. PMID- 8416105 TI - [Long-term effects of inpatient psychotherapy: explanatory and transfer forms of the psychotherapy experience into everyday life]. AB - We describe the longterm effects of inpatient psychotherapy based on biographic interviews with 46 ex-patients two years after their treatment in a psychotherapy ward which is part of a psychiatric hospital. The concept of the treatment is based on cognitive behavior therapy. In a structural hermeneutic analysis of the interviews we found 4 different types of dealing with the experience of psychic disorder and psychological treatment and, combined with it, transfer of the knowledge about psychological disorder and treatment into everyday life: 1 the "development"-type-psychotherapy as a method to reframe the individual development. 2 the "deficit"-type-psychotherapy as a system to learn how to deal with individual deficits, 3 the "deviance"-type-psychotherapy as an instrument to renormalize deviation, 4 the "overburden"-type-psychotherapy as a private area to recover and regenerate. PMID- 8416106 TI - [Psychoneuroimmunologic research in psychosomatic medicine]. AB - In German psychosomatics there appears to exist a selective appraisal of studies investigating the correlations of psychological stress and immunity within the much wider spectrum of psychoneuroimmunological research. This is so despite the fact that so far these studies show contradictory results which are difficult to apply to clinical questions. Immuno-behavioral or immuno-psychological effects are mainly dealt with within a biological psychiatric framework. The network theories referred to in the text do not yield a satisfactory integration as yet. Using the question of mind, behaviour and multiple sclerosis as an example, it can be shown that psychoneuroimmunological research may contribute to empirically testable hypotheses for a better integration of (immune-)biological effects on the mind--without giving in to incompatibly reductionist theories. PMID- 8416107 TI - [Neurosis and religiosity. Is there a causal correlation?]. AB - The interrelation between religiosity, mental health and psychopathology is reviewed on the basis of the current literature. Attributional labels, such as the German term "ecclesiogenic neurosis" are submitted to a critical reevaluation, and their validity as a construct is questioned. Neurotic symptomatology in religious patients has to be seen against the background of their underlying pathology, their biography, and the way in which they integrate religion into their life styles. The religiosity of a patient cannot be singled out as a causative and thus primary pathogenetic factor. Rather religion has to be seen as one forming element among others contributing to the content of a neurotic process. Therapists working with religious patients should try to understand the religious background of their patients in order to help them integrate their religiosity in a larger framework of therapeutic goals. PMID- 8416108 TI - [A map of dyadic relations in the Giessen Test]. AB - A series of random samples was carried out to validate a new model for diagnosing couple relationships using the Giessen test. The model delineates all theoretically possible types of couple relationships in the course of which individual scores can be compared simultaneously with standard scores. The interpretation is based on a two-dimensional diagram, which is to be understood as a reduction of a more complex structure. PMID- 8416109 TI - One voice, one vision--uniting to improve Hispanic-Latino health. PMID- 8416110 TI - Improving access to health care in Latino communities. AB - Public debate about health care reform often focuses on the need for health insurance coverage, but in Latino communities many other barriers also inhibit access to medical care. In addition, basic public health services often go underfunded or ignored. Thus, health care reform efforts, nationally and in each State, must embrace a broader view of the issues if the needs of Latino communities are to be served. This report reviews and summarizes information about the mounting problems Latino communities face in gaining access to medical care. Access to appropriate medical care is reduced by numerous financial, structural, and institutional barriers. Financial barriers include the lack of health insurance coverage and low family incomes common in Latino communities. More than 7 million Latinos (39 percent) go without health insurance coverage. Latinos without health insurance receive about half as much medical care as those who are insured. Structurally, the delivery system organization rarely reflects the cultural or social concerns of the communities where they are located. Therefore, providers and patients fail to communicate their concerns adequately. These communication problems are exacerbated by the extreme shortage of Latino health care professionals and other resources available. Institutional barriers often reflect the failure to consider what it means to provide good service as well as high-quality medical care. Reducing these barriers to medical care requires modifying governmental and institutional policies, expanding the supply of competent providers, restructuring delivery system incentives to ensure primary care and public health services, and enhancing service and satisfaction with care. PMID- 8416111 TI - Improving data collection strategies. AB - The authors examined 21 major health data systems of the U.S. Department of Health and Human Services (DHHS) and concluded that data on Hispanics are not included in several Departmental national health data collection systems, and that even when collected, data on Hispanic subpopulations are found in few of the systems. Of the 21 data systems, 6 do not collect Hispanic population data, including the Medicare statistical system. Only the National Vital Statistics System was found to collect data for all major Hispanic subpopulation groups. Seventeen of the 21 data systems do not collect sample sizes adequate for analyzing any one of the four major Hispanic subpopulation groups. To address that lack, Hispanic health leadership agendas have recommended collecting data on Hispanics in all systems, where possible, to provide samples of a size adequate for detailed analysis of Hispanic subpopulation groups, for support of researchers and a Hispanic research infrastructure, and for broad dissemination of data findings, including dissemination in useful formats to Hispanic community based organizations. PMID- 8416112 TI - The development of a relevant and comprehensive research agenda to improve Hispanic health. AB - The development of an appropriate research agenda for Hispanics requires progress in three areas: (a) developing an appropriate research infrastructure, (b) increasing the availability of appropriate research instrumentation, and (c) identifying and assigning priority areas. In addition, a Latino health research agenda must identify mechanisms for increasing the number of trained Hispanic researchers and the number of Latino professional staff members at the Department of Health and Human Services. It is recommended that an Office of Hispanic Health be established within the Office of Minority Health at the Department to oversee the implementation of the recommendations made as part of the Surgeon General's National Hispanic Health Initiative. PMID- 8416113 TI - Increasing the representation of Hispanics in the health professions. PMID- 8416114 TI - Health promotion and disease prevention strategies. AB - The rapid growth and relative youth of the Latino population of the United States, soon to become the largest ethnic-racial group in the country, presents numerous opportunities and a special challenge to those concerned with the health and well-being of this segment of people. The challenge is to develop and implement efficacious strategies for improving Latino health at a time when the overall socioeconomic profile of this population is in striking contrast to that of the rest of the American people. Much can be done to improve Latino health by implementing health promotion and disease prevention interventions designed to achieve Latino parity in reaching year 2000 national health care objectives. The problems encountered by Latinos in obtaining health promotion and disease prevention services, previous recommendations on how these problems could be addressed, and new strategies for implementing more effective services for Latinos are summarized. PMID- 8416115 TI - Mortality attributed to misuse of psychoactive drugs, 1979-88. AB - To assess mortality attributed to misuse of psychoactive drugs in the United States from 1979 through 1988, the authors obtained from death certificates the annual number of, and age-, sex-, and race-specific data for, deaths in which psychoactive drugs were coded as the underlying or contributing cause. Deaths with psychoactive drugs specified as underlying cause (drug-induced) increased from 6,500 (2.9 per 100,000) in 1979 to more than 10,000 (3.8 per 100,000) in 1988. Deaths with psychoactive drugs specified as either underlying or contributing cause (drug-related) increased from 7,200 (3.2 per 100,000) in 1979 to more than 14,400 (5.5 per 100,000) in 1988. The drugs that primarily accounted for this increase were illicit, in particular, the opiates (heroin) and cocaine, with most of the remainder accounted for by misuse of various legal drugs. The largest increases between 1979 and 1988 occurred among black men ages 35-44 whose drug-induced death rates rose from 8 to 36 per 100,000 and whose drug-related death rates from 10 to 82 per 100,000. These data identify a high-risk group for targeting efforts to prevent deaths due to misuse of psychoactive drugs. PMID- 8416116 TI - Patterns of hospital use by patients with diagnoses related to HIV infection. AB - The authors analyzed the use of hospitals by patients with a diagnosis of human immunodeficiency virus (HIV) infection, using data from the National Hospital Discharge Survey. In the period 1984-90, the rates of both discharges and days of care for HIV-infected patients rose dramatically. For 1988-90, black males had the highest HIV-related discharge rate, followed by white males and black females, whose rates were similar. The discharge rate for patients with HIV related diagnoses increased more in the Northeast than in the three other regions of the country. By 1990 the rate for the Northeast was nearly triple the rate for other major regions. More than half of female and black patients with HIV-related diagnoses were hospitalized in the Northeast. Private insurance was the principal expected source of payment for the care of half of the HIV-infected patients discharged in 1985, but for only a third in 1990. Medicaid covered 40 percent of the patients with HIV-related diagnoses discharged in 1990. Larger proportions of female than male patients and of black patients than white patients were covered by Medicaid. Acquired immunodeficiency syndrome was the diagnosis coded for most patients with an HIV-related diagnosis, but in larger proportions for patients who were male or white patients. Nonspecific HIV diagnoses were coded for larger proportions of female and black patients. HIV-infected patients had an average of 3.6 diagnoses in addition to their HIV diagnosis. Nearly a fourth of the additional diagnoses were for other infectious diseases, such as pneumocystosis or candidiasis. Anemia, pneumonia, and drug use and dependence also were frequent diagnoses. PMID- 8416117 TI - Changes in sexual behavior by young urban heterosexual adults in response to the AIDS epidemic. AB - Young adults residing in four States were enrolled in the period 1985-88 in a multicenter study of cardiovascular disease risk factors. In 1989, 2,729 members of the group were given a self-administered questionnaire that included questions on changes in sexual behavior that subjects had made in response to the epidemic of acquired immunodeficiency syndrome (AIDS). The final sample of 1,601 young, heterosexual, urban respondents included 412 white men, 568 white women, 224 black men, and 397 black women, all ages 21 to 40 years. Overall, nearly 50 percent of the sample reported having made at least one change in their sexual behavior in response to the AIDS epidemic to decrease their risk of becoming infected by the human immunodeficiency virus. The mean number of changes was 0.8 for white men, 1.1 for white women, 1.6 for black men, and 1.5 for black women. Change was reported more frequently by black respondents than white, with no significant sex differences. The categories of respondents reporting behavior changes were more often young, with a history of recreational drug use, with more sex partners, or having had anal intercourse. The most commonly reported behavior changes were reducing the number of sex partners and being more careful in partner selection. Of the 54 percent of respondents who did not report any change in sexual behavior, about 70 percent reported unprotected sex with more than one partner in the previous year. Significant sexual behavior change in response to the AIDS epidemic remains a goal for health education efforts. PMID- 8416118 TI - Estimation of the extent of the cutoff region from the spatial distribution of labelling and mitotic indices of intestinal crypts of a fixed length. AB - Current understanding of the pattern of proliferation within intestinal crypts involves the notion of a cutoff region introduced by Cairnie et al. (Exp. Cell. Res. 39, 539-553, 1965b). (Cells produced above the cutoff are non-cycling, whereas cells produced below the cutoff are cycling.) They contrasted the predicted distribution of proliferation in the extreme cases of a cutoff of width 0 (a sharp cutoff) with one eight cells wide (a slow cutoff) and concluded that the data were better explained by the latter. We have shown that crypt size variation artificially broadens the apparent distribution of proliferating cells in the crypt (Totafurno et al., Biophys. J. 54, 845-858, 1988). Here we show that the measurement and analysis of crypts of a specified height reduces this artifact. This work introduces the use of distance from the crypt base (in microns) to specify the location of cells within the crypt as an improvement over the cell position ordering traditionally used in the determination of the distribution of proliferating cells. We also show how to explicitly correct for several artifacts in the measurement of the labelling index. We conclude that cell proliferation within the crypt is more localized than previously realized; in fact, a cutoff as slow as eight cells wide is rejected. PMID- 8416119 TI - Transesophageal echocardiography in valvular heart disease. PMID- 8416120 TI - Cardiac magnetic resonance imaging. AB - All four cardiac valves can be imaged using MRI. Gradient-echo imaging is the preferred mode of imaging. Valvular regurgitation seen as an area of signal loss in the more proximal chamber can be diagnosed with a high degree of accuracy when compared with 2-D Doppler echocardiography and catheterization angiography. Aortic and mitral stenosis can be semiquantitatively diagnosed, but no method for determining valve areas is currently available. Cardiac prosthetic valves can be imaged but appear only as localized signal loss. Prosthetic valve regurgitation can be diagnosed in the same way as native valve regurgitation. At present, MRI, though not a widely used modality, can contribute significantly to the diagnosis of cardiac valve disorders. With the addition of fast magnetic resonance scanning, which can eliminate the need for electrocardiographic gating, it will be possible for patients with cardiac rhythm irregularities to be scanned, thus broadening the base of patients with valve disease who can be diagnosed. PMID- 8416121 TI - Endocarditis: recognition, management, and prophylaxis. PMID- 8416122 TI - Valvular disease in the elderly. AB - As patients survive to advanced age, they commonly develop degenerative valvular heart disease as well as degenerative diseases of other organ systems. In addition, a reservoir of patients with other forms of valvular heart disease develop progressive symptomatology with advancing age. These patients often present a challenge to the cardiologist in both diagnosis and management. Inasmuch as these patients tolerate cardiovascular surgery less well than their younger counterparts, criteria for surgical intervention may often need modification. Chronologic age must be recognized as but one of many factors affecting physiologic function. Knowledge of aging-related alterations in function must be employed in both diagnostic and therapeutic algorithms. At times, input from other health-care providers who specialize in the care of the elderly may assist in the assessment of these patients. Surgery should be reserved for higher-risk patients who are severely symptomatic or for those in whom severe symptoms are likely to soon develop based on the natural history of the disease process involved. Those less symptomatic elderly patients with otherwise preserved physiologic functions also may be offered valvular surgery. The availability of nonsurgical, albeit at times palliative, techniques to relieve aortic or mitral stenosis provides an alternative therapeutic option to cardiothoracic surgery. Advances in understanding the pathophysiology and medical therapy of heart failure will continue to contribute to an improved quality of life for those for whom only medical options exist. PMID- 8416123 TI - Timing of surgery for valvular heart disease. PMID- 8416124 TI - Mechanical prostheses: old and new. AB - The four mechanical valve prostheses currently available on the U.S. market have evolved from a field of more than 50 valves produced for human implantation since the early 1950s (Tables 11-1 to 11-4). This literature establishes that good results can be achieved with a number of cardiac prostheses if properly used and monitored after implant. The current generation of valves have demonstrated ease of implantation, improved durability, good hemodynamic performance, and reduced thromboembolism and thrombosis with proper anticoagulation. The cost and complexity of completing PMA by the FDA, concern over product liability, and patent rights on design and raw materials have narrowed the choice of devices for surgeons in the United States and slowed the pace of new market entries. The evolution of mechanical valves has been reviewed and modes of valve failure reviewed when pertinent. Clinical expectations for earlier generation devices and present valves also are reviewed. Prostheses under evaluation are discussed along with considerations for valve implantation, surveillance, and anticoagulation. We have employed the SJM valve since about 1985. The proven good hemodynamic performance in small sizes and low profile have made its application well suited to the pediatric population and for smaller aortic roots. The well-guarded hinge mechanism and low probability for disc entrapment have facilitated its use in chordal sparing mitral replacements in our experience. Application in the tricuspid position also has been successful but requires close attention to anticoagulation. PMID- 8416125 TI - Valvular heart disease of congenital origin. PMID- 8416126 TI - Mitral valve reconstruction. PMID- 8416127 TI - Tricuspid valve surgery: indications, methods, and results. PMID- 8416128 TI - Immediate postoperative management. AB - The management of the patient after valvular surgery is influenced by the preoperative left ventricular function and by the specific type of valvular lesion that has been corrected. The hemodynamics of the patient immediately after surgery must be evaluated carefully, and the patient can be supported with a variety of pharmacologic and mechanical measures as ventricular function recovers. Perioperative ischemia, arrhythmias, and bleeding may occur and should be specifically treated. Initiation of anticoagulation and adjustment of warfarin doses is an important aspect of postoperative care. Finally, the patient needs to be counseled that the presence of a prosthetic valve carries a risk for infective endocarditis and instructed on appropriate regimens for antibiotic prophylaxis. PMID- 8416129 TI - Valvuloplasty. PMID- 8416130 TI - Acute rheumatic fever. AB - During the first half of this century rheumatic fever was a common disease with significant morbidity and mortality in the United States. In the 1980s, when many clinicians were hoping this disease was a disease of the past, anxieties were renewed when outbreaks were reported in several areas around the country. Although the etiology still eludes us, insight has been gained. Environmental and genetic factors are believed to play a role in the epidemiology of this disease. Additionally, the implicated organism, the group A streptococcus, has many strains, and differences in its many proteins may determine their potential for rheumatic fever. The mechanisms leading to disease are not clear, but the streptococcus has been implicated as a source of antigens with cross-reactivity with human tissues and has been shown to modify immune mechanisms. Clinical aspects are briefly reviewed and physicians are reminded to consider rheumatic fever as a diagnostic possibility in the appropriate settings. PMID- 8416131 TI - The electrocardiogram in valvular heart disease. PMID- 8416132 TI - Arrhythmias in valvular heart disease. PMID- 8416133 TI - Echo-Doppler in valvular heart disease. AB - Although Doppler echocardiography plays an important role in evaluating patients with suspected valvular heart disease, it should not replace a careful history, a meticulous physical examination, an electrocardiogram, and well-performed posteroanterior and lateral chest x-rays. Two-dimensional echocardiography can reliably evaluate anatomic valvular lesions, estimate left and right ventricular function, and exclude associated pericardial disease. Doppler echocardiography provides accurate hemodynamic parameters of the severity of aortic and mitral stenosis and the degree of pulmonary hypertension. In addition, color-flow Doppler is helpful in providing semiquantitative information with regard to the degree of MR, AR, or TR. Doppler echocardiography is very useful in evaluating patients before and after valvuloplasty but may be inaccurate when compared with cardiac catheterization immediately following mitral balloon valvuloplasty. However, in the long-term followup, after valvuloplasty, Doppler echocardiography is ideally suited to predict restenosis. A properly performed echo-Doppler study may allow the clinician to send a young patient for surgery when warranted by the clinical symptoms. However, in older patients, especially those with suspected coronary artery disease, and in multivalvular disease, cardiac catheterization may still be required. PMID- 8416134 TI - Histological examination of 2859 breast biopsies. Analysis of adequate sampling. PMID- 8416135 TI - A rational approach to the understanding and management of the dysplastic nevus syndrome concept. PMID- 8416136 TI - Non-neoplastic polypoid lesions and adenomas of the gallbladder. PMID- 8416137 TI - Placental pathology for the nineties. AB - Examination of the placenta in a problem pregnancy is potentially very valuable, yielding information about the nature and duration of processes occurring in gestation. Such evaluation requires thoughtful gross examination, careful sectioning, and an understanding of the basic microscopic changes in a variety of pathologic processes. As there are very few absolute associations in placental pathology, it is most important to appreciate the range of normal in assessing the potential importance of lesions. This chapter has tried to provide the essential relevant material, with emphasis on commonly encountered problem areas, and a variety of topics potentially related to maternal and fetal morbidity and mortality. PMID- 8416138 TI - Distribution of C cells in the normal and diseased thyroid gland. PMID- 8416139 TI - Pulmonary aspergillosis: pathologic and pathogenetic features. AB - Pulmonary aspergillosis is a relatively common fungal infection in individuals who are immunocompromised or have intrinsic lung disease. Clinical, radiological, and pathologic manifestations are quite varied and depend to a large extent on the type and severity of local or systemic host defense abnormalities. In individuals with only structural lung damage, saprophytic growth alone is the rule. Patients with atopy or other hypersensitivity state typically develop allergic disease, most often allergic bronchopulmonary aspergillosis. Individuals with other immunologic abnormalities, particularly immunodeficiency, and with granulocytopenia characteristically develop invasive disease, which may take several morphological forms. Identification of Aspergillus as the cause of all these disease variants is usually not a problem. However, recognition of the different patterns of disease is useful in understanding the pathogenesis of disease and in interpreting premortem clinical and radiographic abnormalities. PMID- 8416140 TI - Vascular tumors in children and adolescents: a clinicopathologic study of 228 tumors in 222 patients. AB - Vascular tumors in 222 children and adolescents accounted for 25 percent of all soft tissue tumors in our series of over 900 pediatric cases. Based upon standard histopathologic criteria, 203 (89 percent) were benign, 21 (9 percent) were borderline or indeterminant, and 4 (2 percent) were malignant. The mean age at diagnosis was 12 years; 26 percent (58) of cases were diagnosed in the first year of life and 57 percent (127) in the first decade. The male:female ratio was 1:2. The head and neck (80 cases, 35 percent), extremities (79 cases, 35 percent), and trunk (65 cases, 28 percent) were respectively the regional sites. Seven patients with localized angiomas had more than one lesion; 22 required re-excision for recurrent tumors. Hemangiomas of one type or another accounted for 64 percent and lymphangiomas for 29 percent of the benign group. Capillary hemangioma (73 cases, 32 percent of entire group) was the most common type, followed by lymphangioma (59 cases, 26 percent). Nine patients had diffuse angiomatosis. Borderline tumors comprised 9 percent (21 cases) of cases including ten histiocytoid hemangiomas and 7 hemangiopericytomas. Angiosarcoma (three cases) and Kaposi's sarcoma (one case) constituted the malignant tumors. PMID- 8416141 TI - Caring for People with Physical Impairment. The Journey Back. Formulated by the Committee on Handicaps. Group for the Advancement of Psychiatry. Report No. 135. PMID- 8416142 TI - Overview of surgery: 1993. PMID- 8416143 TI - The current management of varicose and telangiectatic veins. PMID- 8416144 TI - Cardiomyoplasty for end-stage heart failure. PMID- 8416145 TI - Techniques for the treatment of distal hypospadias. PMID- 8416146 TI - Urban bomb blast injuries: patterns of injury and treatment. AB - A review of many series reporting injuries following blasts, data allows certain conclusions to be made: 1. Most patients sustain minor injuries, which may be treated on an outpatient basis. 2. Injuries predominantly affect the head and neck and the periphery, which suggests that clothing plays a major role in protection from secondary injuries. 3. Injuries to the chest and abdomen are relatively uncommon but have a high mortality, also associated with head injury. 4. Primary blast injuries are uncommonly seen in a hospital setting, because they usually result in immediate death. PMID- 8416147 TI - The principles and applications of surgical adhesives. PMID- 8416148 TI - Minimizing perioperative homologous blood exposure with recombinant human erythropoietin. AB - Stimulation of erythropoiesis is an attractive alternative to the risks of homologous transfusion. The availability of rHuEPO has made erythropoietic acceleration possible. The use of rHuEPO perioperatively and in preoperative autologous donation will likely find firm indications. Combinations of present methods of autologous blood use with the various rHuEPO regimens will be the best methods of minimizing perioperative homologous blood exposure. PMID- 8416149 TI - Molecular basis of epithelial tumorigenesis: the thyroid model. AB - Although a minor cause of cancer mortality, thyroid tumors represent a simple and hence powerful experimental model for studying the cell and molecular biology of tumorigenesis in human epithelial cells. This review uses current knowledge of the physiology of growth control in the thyroid as a framework for discussing the somatic genetic abnormalities responsible for follicular cell tumors. Specific emphasis is placed on the predictable involvement of the G-protein oncogene gsp, the key early role of the ras oncogene family, and the apparent rarity of mutations in the p53 tumor-suppressor gene. Potential contributions of the thyroid model to our understanding of interactions between growth regulatory genes are discussed, particularly the relationships between ras and IGF-1 and between p53 and TGF-beta. Throughout, thyroid tumor data are related to that from other tumor types and interpreted in the context of a general model of cell proliferation. PMID- 8416150 TI - The epidemiology of thyroid carcinoma. AB - Thyroid cancer is one of the rarest forms of cancer, yet there are wide variations in the degree of malignancy, ranging from the most rapidly fatal to the relatively benign. This is almost entirely dependent on the histological type. Generally speaking, data available on changing trends of incidence and mortality are subject to reservation, dependent on the degree to which they have been influenced by changing diagnostic criteria and the precision of histopathological description. Nevertheless, there is evidence that mortality is slowly decreasing, while incidence is slowly increasing. The purpose of this review is to try to interpret the temporal trends of incidence and mortality rates for thyroid carcinoma in the last 3 decades in light of the problems arising from changes in diagnostic standards and histological classification regarding thyroid neoplasms. Attention is also drawn to the implications, from a public health viewpoint, of the present intensive detection and treatment of occult thyroid carcinomas. PMID- 8416151 TI - Impact of the cholesterol education program for nurses: a pilot program evaluation. PMID- 8416152 TI - Effect of fecal occult blood testing on mortality from colorectal cancer. A case control study. AB - OBJECTIVE: To estimate by case-control methods the effect of screening using the fecal occult blood test (FOBT) on mortality from colorectal cancer and to examine the relation of that effect to the interval since the most recent screening test. DESIGN: A case-control study. SETTING: The Kaiser Permanente Medical Care Program of Northern California. PATIENTS: A total of 485 persons who developed fatal colorectal cancer after 50 years of age and 727 age- and sex-matched controls. MEASUREMENTS: History of screening FOBTs during the 5 years before case diagnosis. RESULTS: After adjustment for potentially confounding factors, an odds ratio of 0.69 (95% Cl, 0.52 to 0.91) was observed for exposure to at least one screening FOBT during the 5-year interval. The odds ratio was lowest for the first year after the most recent FOBT and rose to 1.00 three years after the last screening examination. False-negative results among cases in the 1 to 2 years before diagnosis contributed substantially to lowering the estimate of efficacy. CONCLUSIONS: These data suggest that a program of annual or biennial screening using FOBTs might lower population risk for mortality from colorectal cancer sufficiently to have important public health implications. However, the confidence intervals around our odds ratio estimates were wide. We therefore believe that additional data will be needed before making recommendations that FOBT screening be expanded. PMID- 8416153 TI - Predicting the occurrence of adverse events after coronary artery bypass surgery. AB - OBJECTIVE: To determine whether adverse events occurring after coronary artery bypass surgery in Medicare patients can be predicted from clinical variables representing illness severity at admission. DESIGN: Retrospective analysis of clinical data abstracted from hospital charts, with development and validation using half-samples of the database. A logistic model was developed using illness severity at admission to predict the occurrence of an adverse event after bypass surgery. SETTING: Hospitals in seven states. PATIENTS: Random sample of 2213 Medicare patients 65 years of age or more who underwent bypass surgery between January 1985 and June 1986. OUTCOME MEASURE: The occurrence of death within 30 days of admission or any of 13 nonfatal postoperative adverse events (for example, myocardial infarction, congestive heart failure, and wound infection). RESULTS: Thirty-three percent of patients had one or more postoperative adverse events or died within 30 days of admission. Mortality within 30 days of admission was 6.6%; each adverse event was associated with increased mortality (range, 7.5% to 66.7%). Admission predictors of the occurrence of an adverse event included a history of bypass surgery, emergent surgery, a history of chronic obstructive pulmonary disease, the presence of an infiltrate on admission chest radiograph, a pulse of 110 beats/min or more, age, blood urea nitrogen of 10.7 mmol/L (30 mg/dL) or more, acute myocardial infarction at admission, and a history of myocardial infarction; the presence of one- or two-vessel disease was negatively associated with the occurrence of an adverse event. The model c-statistic was 0.64. CONCLUSIONS: Severity of illness at admission has modest predictive power with respect to adverse-event occurrence in Medicare patients who undergo bypass surgery. PMID- 8416154 TI - Limitations of impedance plethysmography in the diagnosis of clinically suspected deep-vein thrombosis. AB - OBJECTIVE: To re-evaluate the accuracy of impedance plethysmography (IPG) for the detection of proximal deep-vein thrombosis (DVT). PATIENTS: A total of 384 of 390 consecutive outpatients referred with their first episode of clinically suspected deep-vein thrombosis. SETTING: University-based tertiary care medical center associated with a cancer clinic. DESIGN: A retrospective analysis of a cohort of patients whose data were recorded and stored prospectively on a computerized data base over a 22-month period. MEASUREMENTS: Patients were evaluated by a physician and underwent IPG testing. Patients with abnormal IPG tests and those with normal IPG results in whom there was a high clinical suspicion of DVT or in whom follow up IPG testing was not feasible were referred for venography. Venography and IPG results were interpreted by a panel of independent observers. Two models of the IPG instrument were used (Codman 200 and Electrodiagnostic Instruments 800). RESULTS: Venography (or compression ultrasound) was done in 57 patients with an abnormal IPG test and in 85 patients with normal IPG results. Impedance plethysmography was abnormal in only 37 of 56 patients with confirmed proximal vein thrombosis (sensitivity, 66%; 95% Cl, 52% to 78%). Of the 57 patients with an abnormal IPG result, 37 had DVT (positive predictive value, 65%). The sensitivity for the detection of proximal DVT did not differ between the IPG 200 and 800 instruments (sensitivity, 63% and 71%, respectively; P > 0.2). Of the 19 proximal-vein thrombi not detected by IPG, 12 (63%) were occlusive and 11 (58%) involved at least the popliteal and superficial femoral veins. CONCLUSIONS: At our center, IPG has a far lower sensitivity for proximal-vein thrombosis than has been previously reported for symptomatic outpatients. The reason for this low sensitivity is unclear. Our findings indicate that centers using IPG as the initial diagnostic test for suspected DVT should be aware of this potential problem and should consider re-evaluating the sensitivity of their IPG machines by performing venography in a cohort of their patients with normal test results. PMID- 8416155 TI - High-dose ifosfamide is associated with severe, reversible cardiac dysfunction. AB - OBJECTIVE: To determine the incidence and characterize the occurrence of cardiac toxicity with high-dose ifosfamide. DESIGN: Retrospective chart review. SETTING: Biomedical research referral center. PATIENTS: Fifty-two consecutive patients with advanced lymphoma or carcinoma enrolled in phase I trials of high-dose ifosfamide as part of combination chemotherapy with autologous bone marrow transplantation. INTERVENTIONS: Patients were given escalating doses (10 to 18 g/m2) of ifosfamide in combination with carboplatin and etoposide or with lomustine and vinblastine. MEASUREMENTS: The chart review focused on clinical, radiographic, or electrocardiographic evidence of cardiovascular dysfunction. Data from invasive hemodynamic monitoring, radionuclide cineangiography, and echocardiography were also reviewed. RESULTS: Nine of the patients treated with ifosfamide developed congestive heart failure (17%; 95% Cl, 8% to 30%). Eight of these patients, experiencing dyspnea, tachycardia, weight gain, and signs of pulmonary edema, required admission to an intensive care unit. Left ventricular contractility was found to be depressed when evaluated by radionuclide cineangiography, echocardiography, or both. Most patients responded to diuretic, vasodilator, and inotropic therapies. Two patients developed malignant ventricular arrhythmias. One patient died of intractable cardiogenic shock. Five patients died of multiorgan failure, despite showing improvement in left ventricular ejection fraction. Three patients survived and regained baseline left ventricular ejection fraction. CONCLUSIONS: High-dose ifosfamide is associated with severe but usually reversible myocardial depression and malignant arrhythmias. PMID- 8416156 TI - Lactic acidosis complicating the acquired immunodeficiency syndrome. PMID- 8416157 TI - Objective evaluation of endoscopy skills during training. AB - OBJECTIVE: To evaluate the number of supervised gastrointestinal endoscopic procedures required to achieve initial competency using a simple objective grading system. DESIGN: Prospective, cross-sectional study. SETTING: A gastroenterology and surgical training program at a large, university-affiliated county hospital. PARTICIPANTS: Seven gastroenterology fellows and five fourth year surgery residents. INTERVENTIONS: Trainees were graded postprocedure using a microcomputer program. Grading criteria for esophagogastroduodenoscopy included entering the esophagus (esophageal intubation), traversing the pylorus into the duodenum, and recognizing whether the upper gastrointestinal tract was abnormal. Criteria for colonoscopy were traversing the splenic flexure, intubating the cecum, and recognizing whether the colon was abnormal. RESULTS: When presented with a case mix representative of practice, esophageal intubation did not reach 90% until more than 100 procedures had been done. Cecal intubation remained at only 84% after 100 procedures. CONCLUSIONS: More than 100 supervised upper gastrointestinal endoscopies or colonoscopies are necessary to achieve technical competence in gastrointestinal endoscopy. PMID- 8416158 TI - The history of medicine. An annotated list of key reference works. PMID- 8416160 TI - Are we always? When do we stop? PMID- 8416159 TI - Malignant polyps: are they sheep in wolves' clothing? AB - One of the arguments supporting the concept that benign adenomatous polyps of the colon degenerate into cancer is the observation of malignancy in polypoid growths known to have been present for years. Although a latent phase--the duration of time between the initial development of a malignancy and the subsequent occurrence of a clinical problem--must exist, this argument implies that this time span is not many years. If both the prevalence of malignant polyps and the incidence of consequent symptomatic colon cancer were known, the average latent phase could be calculated. In order to estimate this prevalence, I used autopsy data; the estimate was validated using independent data fro three colonoscopic screening studies. The annual incidence of all colon cancer in the United States is approximately 150,000 cases. I estimate that 725,000 people in the United States harbor at least one malignant polyp. Even if all 150,000 cases of colon cancer were associated with symptoms and began as malignant polyps, the average latent phase is 4.8 years. Because some colon cancers are removed while the patient is still asymptomatic (discovered on screening examination) and because at least some colon cancers arise de novo from the mucosa, the average latent phase must be even longer. These estimates suggest that it cannot be assumed that the histologic finding of cancer in a polyp that has been observed for many years represents "malignant degeneration" of a previously benign neoplasm; such a malignancy may have had that histologic characteristic from the start. Further, histologically ominous lesions (malignant polyps) may often have prolonged benign clinical courses. PMID- 8416161 TI - Tolrestat for mild diabetic neuropathy. A 52-week, randomized, placebo-controlled trial. AB - OBJECTIVE: To evaluate the effectiveness and safety of tolrestat, an aldose reductase inhibitor, in patients with mild diabetic autonomic and peripheral neuropathy. DESIGN: Randomized, placebo-controlled, double-blind 52-week trial. SETTING: University hospital clinic. PATIENTS: Forty-five diabetic patients with asymptomatic autonomic neuropathy identified by at least one pathologic cardiovascular reflex test result. INTERVENTIONS: All patients were given placebo during a 4-week run-in period (single-blind). Twenty patients were randomly assigned to continue to receive placebo, and 25 were assigned to treatment with tolrestat (200 mg/d given in the morning). MEASUREMENTS AND RESULTS: At 12 months, improvements in nerve functions occurred in patients receiving tolrestat. Compared with baseline values, postural hypotension decreased by a value of 5.9 mm Hg (95% Cl, 1.6 to 8.7); deep-breathing, maximum/minimum heart rate (expiration/inspiration ratio) increased by a value of 0.026 (Cl, 0.015 to 0.036); and lying-to-standing heart rate ratio (30:15 ratio) increased by a value of 0.032 (Cl, 0.027 to 0.052). In the placebo group, all test results except postural hypotension deteriorated. Vibration perception threshold at the malleolus and great toe of the dominant leg improved in the tolrestat group ( 1.4; Cl, -3.69 to -1.09) but tended to worsen in the placebo group during the study period. No important side effects were detected in either group. CONCLUSIONS: The progression of mild diabetic autonomic and peripheral neuropathy may be halted or even reversed by pharmacologic intervention with the aldose reductase inhibitor tolrestat. PMID- 8416162 TI - Colonic polyps, cancer, and fecal occult blood. PMID- 8416163 TI - On endoscopic training and procedural competence. PMID- 8416164 TI - Eliciting patient preferences. PMID- 8416165 TI - Clusters of multidrug-resistant tuberculosis. PMID- 8416166 TI - The dismantling of tuberculosis control programs. PMID- 8416167 TI - Calculation errors in meta-analysis. PMID- 8416168 TI - Reflections on the doctor's anguish. PMID- 8416169 TI - Reflections on the doctor's anguish. PMID- 8416170 TI - Reflections on the doctor's anguish. PMID- 8416171 TI - Reflections on the doctor's anguish. PMID- 8416172 TI - Reflections on the doctor's anguish. PMID- 8416174 TI - Moving points in nephrology. Festschrift in honor of Geoffrey M. Berlyne on the occasion of his 60th birthday. PMID- 8416173 TI - Reflections on the doctor's anguish. PMID- 8416175 TI - The hyperkalemia of renal failure: pathophysiology, diagnosis and therapy. PMID- 8416176 TI - Aluminum and renal disease. PMID- 8416177 TI - The use of desferrioximine in dialysis-associated aluminium disease. PMID- 8416178 TI - Effects of chronic renal failure with and without excess parathyroid hormone on calmodulin content of pancreatic islets and on the response of their Ca2+ ATPase to calmodulin. PMID- 8416179 TI - How is urine concentrated by the renal inner medulla? PMID- 8416180 TI - Renal function and water metabolism in the dromedary. PMID- 8416181 TI - Water immersion: lessons from antiquity to modern times. PMID- 8416182 TI - The kidney in familial Mediterranean fever. PMID- 8416183 TI - Ischaemic heart disease, lipids and the kidney. PMID- 8416184 TI - Nephrology and the rationing of health care. PMID- 8416185 TI - Acute renal failure: a multifactorial syndrome. Pathogenesis and prevention strategies. AB - Professor Geoffrey M. Berlyne has devoted many years of his productive life to improve our understanding of kidney physiopathology and to enhance our treatment modalities for renal failure. This review of acute renal failure, with emphasis on some recent progress in our understanding of this syndrome, is dedicated to honor Professor Berlyne's 60th birthday. PMID- 8416186 TI - Perspectives on dialysis in the Third World: a problem of economics. PMID- 8416187 TI - Diet and chronic renal failure. PMID- 8416188 TI - Toxins versus abnormal cell metabolism in the pathophysiology of chronic renal failure. PMID- 8416189 TI - A case study of a recent decline in the dialysis fatality rate. AB - Geoffrey Berlyne, whom we honor in this Festschrift, has contributed much to the modern understanding of diseases of the kidney, and of the clinical and metabolic disorders that occur in acute and chronic renal failure. The Festschrift highlights the many areas of his contributions. It is important to note that underlying them all is astute clinical observation, incisive analysis and reasoning, a breadth of approach, experimentation that facilitated understanding of the relevant clinical issues, and an unusual clarity of exposition in the literature. As author, editor, and teacher, Geoffrey Berlyne has brought this clarity of approach to a generation in nephrology. The following analysis of clinical data from a working dialysis unit, rendering care to a predominantly underprivileged patient population, is presented in the hope that it throws light on everyday problems in nephrology in a manner similar to that in which Geoffrey has guided the nephrology community. PMID- 8416190 TI - High-flux hemodialysis: overcoming the tyranny of time. PMID- 8416191 TI - Recent perspectives on peritoneal dialysis. PMID- 8416192 TI - Analysis of anti-Tax antibody of HTLV-I carriers in an endemic area in Japan. AB - Sera from 1197 adult residents in Miyazaki district, an area in Japan endemic for human T-cell leukemia virus type I (HTLV-I), were tested for anti-Tax antibody by the recombinant Tax (r-Tax) Western blot assay. Among HTLV-I-seropositive individuals, including 21.5% of 484 males and 28.6% of 713 females, the prevalence of anti-Tax antibody were 59.6% and 58.3% respectively, with no apparent difference in age. There was a significant 6-fold difference in the prevalence of anti-Tax among seropositive subjects with titer > or = 1:8192 (84.6%) compared with those with the lowest titer of 1:16 (14.3%), suggesting the increased production of antibodies to viral structural proteins in anti-Tax positive individuals. Furthermore, among those anti-Tax-positive subjects, the intensity of serum reactivity to r-Tax protein in the high antibody titer (1:1024 or higher) group was significantly stronger than that in the lower antibody titer (1:512 or lower) group. We also found that 1.6% (14/889) of individuals without detectable levels of HTLV-I antibody had anti-Tax antibody. HTLV-I pro-viral DNA signals could not be detected in DNA sample from the lymphocytes of these individuals by the nested polymerase chain reaction method. Further evaluation is needed to clarify the significance of an anti-Tax-only status population in which HTLV-I is endemic. PMID- 8416193 TI - Gangliosides and organ-specific metastatic colonization. AB - To investigate whether metastatic specificity is associated with a variation in the ganglioside profile of the parent cell lines, we have analyzed and compared the ganglioside content of subcutaneous (s.c.) tumors to that of the corresponding metastases. C57BL/6 mice were injected with either the Lewis lung carcinoma (3LLc) or its cloned variants, M27, exclusively metastatic to the lung, and H59, metastasizing preferentially to the liver. Gangliosides were extracted, purified and separated on HPTLC. H59 liver metastases contained significantly more GM2 (27.8%) than the H59 s.c. tumor (7.6%). The ganglioside profiles of 3LLc or M27 s.c. tumors were no different from those of their corresponding metastases. GM2 predominated in the liver (90.8%) while GM3 (48.8%) and GM2 (33.8%) were prevalent in the lung. Unidentified gangliosides designated G1, G2, G4 and G5 were present in tumor cells but absent from normal lung and liver. This study indicates that the ganglioside compositions of the 3LLc cell line and of its M27 variant were not modified under the influence of different sites of growth. Furthermore, the ganglioside profiles of the metastases were distinct from those of their respective target organs. The results of these studies suggest a possible relationship between GM2 and the establishment of H59 metastases in the liver. PMID- 8416194 TI - Loss of heterozygosity at chromosome 17p is associated with HER-2 amplification and lack of nodal involvement in breast cancer. AB - Sixty DNA samples from breast carcinoma (BC) patients were analyzed by Southern blot to examine certain oncogene and anti-oncogene alterations. Amplification of the HER-2 oncogene was detected in 15 tumours (25%), c-myc in 2 (3%) only and HER 1 in none. Distribution of Hras-1 oncogene alleles in BC did not significantly differ from that seen in healthy donors. Not a single case of RB-1 anti-oncogene alteration and only one of p53 suppressor gene abnormality was found when appropriate cDNA copies were used as probes. With the use of DNA polymorphic markers, loss of heterozygosity (LOH) was revealed at chromosome 17p (probe YNZ 22) in 18 of 39 (46%) informative cases, at 17q (probe THH-59) in 10 of 34 (29%) and at 11p (probe Hras-1) in 8 of 30 (27%). The only significant correlation between these genetic alterations and the clinical characteristics of the tumours studied was the association of LOH at 17p with node-negative BC (p < 0.02). Also, the tendency for correlation (p < 0.2) between HER-2 amplification and loss of 17p sequences was revealed: 7 of 10 amplification-positive, but only 11 of 29 amplification-negative BC possessed LOH of YNZ-22 locus. PMID- 8416195 TI - Interaction of topoisomerase I inhibitors with radiation in cis diamminedichloroplatinum(II)-sensitive and -resistant cells in vitro and in the FSAIIC fibrosarcoma in vivo. AB - The cytotoxicity of the topoisomerase-I inhibitors, camptothecin and topotecan, toward the SCC-25 human head-and-neck squamous-carcinoma cells and the SCC 25/CDDP sub-line made resistant to cis-diamminedichloroplatinum(II) was assessed alone and in combination with radiation. Topotecan was less cytotoxic than camptothecin in cell culture and the SCC-25/CDDP cell line was more sensitive to either topoisomerase-I inhibitor than was the parental SCC-25 cell line. Both camptothecin and topotecan were effective radiation sensitizers of hypoxic SCC-25 and SCC-25/CDDP cells under normal pH or acidic pH conditions. Sensitizer enhancement ratios ranged between 1.5 and 1.6 for hypoxic SCC-25 cells and between 1.3 and 1.5 for hypoxic SCC-25/CDDP cells. When the ability of camptothecin or topotecan to sensitize the FSallC fibrosarcoma to single-dose radiation was assessed using the tumor-cell-survival assay, a sensitizer enhancement ratio of 1.2 was found with each drug. However, using tumor growth delay of the FSaIIC fibrosarcoma to determine the effect of camptothecin or topotecan to enhance the efficacy of a daily fractionated radiation regimen, topotecan produced a sensitizer-enhancement ratio of 1.4, while that for camptothecin was 1.2. These results indicate that topoisomerase-I inhibitors may retain activity in CDDP-resistant cells and may be effective adjuncts to radiation therapy. PMID- 8416196 TI - Induction of apoptosis in human leukemic cells by the ether lipid 1-octadecyl-2 methyl-rac-glycero-3-phosphocholine. A possible basis for its selective action. AB - Ether-linked glycerophospholipids (ether lipids, EL) are membrane-interactive drugs selectively cytotoxic toward neoplastic cells compared with normal cells. No conclusive explanation has yet been provided for this selectivity. We now present data indicating that the drug 1-octadecyl-2-methyl-rac-glycero-3 phosphocholine (ET-18-OMe) induces apoptosis, or programmed cell death, in human leukemic cells. Apoptotic death is induced selectively by ET-18-OMe in HL60 cells, which are sensitive to the drug's cytotoxic action, but not in the resistant K562 cell line. Enrichment of HL60 cells with cholesterol (HL60-CHOL cells) significantly protects the cells from the cytotoxic effect and from the induction of apoptosis by ET-18-OMe; the percentage of fragmented DNA is only 17% for HL60-CHOL, compared with 50% in native HL60 cells after exposure to 20 microns ET-18-OMe for 24 hr. Our study provides a possible explanation for differences in sensitivity to EL among different cell types and illustrates an indirect interaction of EL with cellular DNA. PMID- 8416197 TI - Macrophage-induced cytotoxicity and anti-metastatic activity of a 43-kDa human urinary protein against the Lewis tumor. AB - Resident, inflammatory or bone-marrow macrophages from C57Bl/6 mice incubated in vitro with a pure human urinary protein (HGP.43) decreased the growth rate of Lewis tumor cells (3LL). This inhibition of 3LL growth was the result of a cytotoxic activity of these macrophages which was independent of oxygen metabolites and nitrous oxide. Murine monoclonal antibodies (MAbs) against HGP.43 inhibited macrophage-mediated cytotoxicity. This cytotoxic activity was not due to the release of cytotoxic factors in the culture supernatant, showing that a contact between macrophages and tumor cells was required to express cytotoxicity. The presence of HGP.43 was absolutely necessary during the incubation of macrophages with target cells. In vivo, in HGP.43-treated mice, the growth of the primary tumor was not delayed but the size and number of lung metastases were significantly reduced 21 days after tumor inoculation. PMID- 8416198 TI - Induction of the fundic mucosa-specific glycolipid with dimethylformamide in gastric-cancer cell lines. AB - We have previously reported that a glycolipid GalNAc beta 1-4[NeuAc alpha 2-3]Gal beta 1-4GlcNAc beta 1-3Gal beta 1-4Glc-Cer named NGM-1 is present specifically in the human gastric fundic mucosa, but not in other organs. In gastric-cancer tissue and cancer cell lines, this glycolipid completely disappears. These findings imply that NGM-1 is expressed only in well-differentiated fundic mucosa. The purpose of this study is to examine the expression of NGM-1 as a differentiation-related molecule. A gastric cell line AZ521 was cultured in the medium with various reagents which had been reported to induce differentiation in cancer cells. The growth of AZ521 was suppressed by the addition of 0.8% dimethylformamide (DMF) to the medium, but not by addition of dimethylsulfoxide (DMSO), retinoic acid or butyric acid. In the ganglioside fraction of the cells cultured with DMF, a glycolipid regarded as NGM-1 which had not been present before treatment was detected using a monoclonal antibody. Suppression of the proliferation of AZ521 by eliminating the serum from the medium could not induce the expression of NGM-1. A colonic cell line treated with DMF also failed to express the glycolipid. The synthase activity of NGM-1 was elevated in the AZ521 cells treated with DMF, but not with DMSO. These results demonstrate that the expression of NGM-1 is induced by DMF specifically in gastric-cancer cells, and suggest the possibility that NGM-1 is a differentiation-related molecule. PMID- 8416199 TI - Effect of drug-light interval on photodynamic therapy with meta tetrahydroxyphenylchlorin in malignant mesothelioma. AB - The influence of the time interval (TI) between drug administration and laser activation on selectivity of meta-tetrahydroxyphenylchlorin(mTHPC)-mediated photodynamic therapy (PDT) for tumour tissue was assessed in BALB/c nude mice bearing human malignant mesothelioma xenografts. Following i.p. administration of 0.3 mg/kg mTHPC, a light dose of 10 J/cm2 and 0.1 W/cm2 was delivered at 650 nm on the tumour and an equal-sized area of the hind leg after 4, 12, 24 and 36 hr and 2, 3, 4, 5 and 6 days to groups of 6 animals (surface irradiance). Then, 72 hr after light delivery, the depth of necrosis was measured in the tumour and in the skin and underlying muscle of the hind leg. Photosensitized necrosis occurred in normal tissue at TI from 4 hr to 3 days and in the tumour at TI from 12 hr to 4 days. The therapeutic ratio of mTHPC-PDT varied significantly with the time interval between drug administration and laser activation and was greatest at an interval of 3 days. mTHPC concentration was measured in 3 control unirradiated animals at all time points in normal tissues and in tumour tissue, and found to be the same in both tissues. Thus the tissue concentration of mTHPC was of limited use as regards the prediction of photosensitizing effects in the tumour model. PMID- 8416200 TI - Production and characterization of a mouse/human chimeric antibody directed against human neuroblastoma. AB - Hybridoma CE7 produces a murine antibody (gamma 1/kappa) which binds to a 190-kDa cell-surface glycoprotein of human neuroblastoma. Because of its tumor specificity, it has been used routinely in clinical pathology to confirm diagnosis of neuroblastoma. We have isolated the gene segments coding for the variable regions of the immunoglobulin H and L chain of this hybridoma. These V genes were used to construct mouse/human chimeric H and L chain genes (gamma 1/kappa) which were then expressed in SP2/0 cells. A cell-binding inhibition assay showed that the specificity of the chimeric CE7 antibody (chCE7) is identical to that of the original CE7. Radioiodinated chCE7 binds to approximately 43,000 sites per neuroblastoma cell with an affinity of 10(10) M-1. In neuroblastoma-bearing nude mice, biodistribution studies with [125I]chCE7 were performed and tumor accumulations of up to 32% of injected dose/g tissue together with low blood and organ uptake were found. PMID- 8416201 TI - Epstein-Barr virus (EBV) nuclear-antigen-2-induced up-regulation of CD21 and CD23 molecules is dependent on a permissive cellular context. AB - The Epstein-Barr virus (EBV) induces unlimited growth of B lymphocytes in vitro, a phenomenon known as immortalization. The elucidation of the mechanisms by which EBV de-regulates B-cell proliferation in vitro will permit an understanding of how the virus contributes in vivo to the genesis of Burkitt's lymphoma (BL) and of lymphoproliferations in immunosuppressed patients. At present, no single EBV immortalizing gene has been identified, and the hypothesis has been made that many viral genes cooperate in establishing an autocrine loop of secretion leading to immortalization. Constitutive expression of B-cell surface molecules such as CD21 and CD23, specifically implicated in the control of B-cell proliferation, is indeed induced at the surface of immortalized B lymphocytes. The expression of the viral nuclear antigen 2 (EBNA2) has been shown to be in part responsible for CD21 and CD23 up-regulation, and EBNA2 is suspected to be a transactivator of cellular genes, although this point remains to be demonstrated. The role of EBNA2 gene, independently of other viral genes, has been investigated by transfection into B-lymphoma lines, but conflicting results have been reported. To further investigate its role in the regulation of CD21 and CD23 molecules, we have compared the effects of EBNA2 expression in 2 sets of B-lymphoma lines infected with P3HR1 EBV strain, and/or transfected with EBNA2 gene. We report here that: (i) EBNA2 expression is not a sufficient condition to induce CD21 and CD23 upregulation, EBNA2's effects are highly dependent on the cellular context, and moreover can be modified by infection with P3HR1 virus; (ii) EBNA2 induces activation of CD23 expression in a very particular way, namely, an increased quantity of CD23 steady-state RNA coding for the form A of the protein, which is not detectable at the cell surface but directly secreted. PMID- 8416202 TI - Increased expression of p34cdc2 and its kinase activity in human gastric and colonic carcinomas. AB - We examined the expression of p34cdc2 and its kinase activity in human gastric and colonic carcinoma cell lines and carcinoma tissues and studied its relation with a tumor-suppressor gene product, p53. All the gastric and colonic cancer cell lines expressed p34cdc2 and showed its kinase activity at various levels. When the cells were arrested in mitotic metaphase by the use of nocodazole, p34cdc2 kinase activity was induced and p53 was apparently phosphorylated. Of 12 gastric carcinoma cases, 11 (91.7%) showed higher p34cdc2 kinase activity in tumor tissues than in corresponding non-neoplastic mucosa. The protein kinase activities in the individual cases were well correlated with the levels of p34cdc2 protein expression. A good correlation was also found between the expression of p34cdc2 and proliferating cell nuclear antigen (PCNA). Almost all the colonic carcinomas showed higher cdc2 kinase activity and increased p34 expression when compared with non-neoplastic mucosa. Interestingly, most of the gastric and colonic carcinomas having high cdc2 kinase activity expressed high levels of p53. These findings suggest that the increased p34cdc2 kinase activity might cause the development and proliferation of gastric and colonic carcinomas, partly through abnormal p53 accumulation. PMID- 8416203 TI - Flow cytometry in comparison with mitotic index in predicting disease outcome in transitional-cell bladder cancer. AB - The DNA content and S-phase fraction were measured by flow cytometry in 448 tumour biopsy specimens from transitional-cell bladder cancer (TCC). The samples were also analyzed for mitotic index, WHO grade and papillary status, and histological and flow cytometric data were then correlated to clinical behaviour of tumours during a mean follow-up period of 9.9 years. TNM classification, WHO grade, papillary status, mitotic index, DNA ploidy and S phase fraction were significantly interrelated. Twenty-four percent of tumours showed heterogeneous DNA indices when measured from multiple samples (measured in 94 cases). Of the histological parameters, independent predictors of progression in superficial tumours were the S-phase fraction and mitotic index. In superficial tumours, S phase fraction and the mitotic index included all the available independent prognostic information in survival analysis, whereas in muscle-invasive tumours T category was the most important prognostic factor. The results suggest that DNA ploidy has no independent prognostic value in transitional-cell bladder cancer, whereas proliferation indices (SPF, mitotic index) are important prognostic factors. Accordingly, malignancy classification of papillary bladder tumours can be based on proliferation indices alone. Nodular tumours run an unfavourable course and their malignancy grading by flow cytometry or by mitotic index is not relevant. PMID- 8416204 TI - Non-Hodgkin lymphoma in Gabon and its relation to HTLV-I. AB - A case-control study was performed in Libreville, Gabon, to determine whether a relationship can be established between the relatively high proportion of non Hodgkin lymphoma (NHL) among all cancer cases and the high seroprevalence rate of HTLV-I observed, and to discover whether cases of adult T-leukemia/lymphoma (ATLL) related to HTLV-I exist in Gabon. From November 1987 to April 1989, a total of 32 patients with NHL were recruited; 6 were infants with Burkitt's lymphoma and 26 were adults with NHL. Each patient was matched with 2 asymptomatic controls for age, sex and ethnic group. HTLV-I serology was done by ELISA and Western blot. Comparison of the groups was done by chi-square analysis. None of the 6 infants with Burkitt's lymphoma and none of their controls had antibodies to HTLV-I. Of the 26 patients with NHL, 7 (26.9%) had HTLV-I antibodies. Among the 52 controls, the HTLV-I rate was 13.4% (n = 7). There was no difference between cases and controls (Fisher's exact test, p = 0.16). Among the 26 NHL, 4 cases fitted the criteria of ATLL and were HTLV-I-positive; 3 others who were positive for HTLV-I were a woman with lymphoblastic gastric NHL and 2 old men with an unclassified lymphoma. From the results of this limited series it is not possible to state that there is an association between NHL and HTLV-I infection. Nevertheless, cases of ATLL related to HTLV-I are reported from this area. Based on the HTLV-I seroprevalence rates reported in Gabon, the estimated incidence rate of ATL among seropositive people in Gabon appears much lower than in Japan. Different explanations can be proposed, but under-diagnosis of ATLL is probably one of the main factors. PMID- 8416205 TI - A sub-site-specific analysis of the relationship between colorectal cancer and parity in complete male and female Norwegian birth cohorts. AB - Register and census data for complete cohorts of Norwegian men and women born between 1935 and 1969 have been used to examine the relationship between reproductive factors and the incidence of colorectal cancer. Among 1.1 million men and 1.1 million women under observation, 491 male and 859 female cases of colorectal cancer were diagnosed during the period of follow-up. Our hazard model estimates clearly show that in these young cohorts, women with 2 or more children run a lower risk of having a malignant tumor in the cecum or ascending colon than do other women of the same age and in the same birth cohort. No association with parity is found with respect to cancer in the transverse or descending colon, whereas a downward trend in the effect estimates, followed by an upturn, appears for the rectum, sigmoid colon and rectosigmoid junction. Such correlations, which also hold when we control for education and place of residence, are not found for men. The observed relationship between parity and cancer incidence is not likely to be exclusively explained by life-style differentials. Presumably, there is a biological effect of the number of pregnancies and deliveries, net of age at first birth and other reproductive factors, on the development of colorectal cancer. PMID- 8416206 TI - Fundal atrophic gastritis as a risk factor for gastric cancer. AB - The role of atrophic gastritis of the gastric corpus (fundal atrophic gastritis) as a high-risk factor was investigated by studying operative findings and follow up data on 690 patients with benign gastric diseases recorded at the Osaka Cancer Registry. The extent of fundal atrophic gastritis was determined by the endoscopic Congo red test. The patients were followed-up from the time of endoscopic examination (1968 to 1976) to December 31, 1987. The vital status of 654 patients (94.8%) at the end of the observation period was determined. During the follow-up period, 22 patients were found to have gastric cancer. The extent of fundal atrophic gastritis was shown to be closely related with the risk of developing gastric cancer. Patients who had been diagnosed as having severe fundal atrophic gastritis showed significantly higher risk of gastric cancer than patients who had been diagnosed as having little or no fundal atrophic gastritis (5.76-fold, calculated with adjustments for age, sex and the follow-up period). A positive linear relationship was found between the risk of developing gastric cancer and the extent of fundal atrophic gastritis. The observed number of gastric cancers was compared with the expected number calculated from the incidence in Osaka Prefecture. Analysis of the results showed that the observed and expected numbers of gastric cancers in patients with severe fundal atrophic gastritis were 11 and 4.8, respectively, the ratio of observed to expected numbers being 2.3 (p < 0.05). These findings indicate that severe fundal atrophic gastritis is a major risk factor for gastric cancer. PMID- 8416207 TI - Interferon-gamma modulates retinoblastoma gene mRNA in monocytoid cells. AB - To study the effect of interferon gamma (IFN-gamma) on the expression of the retinoblastoma (RB) susceptibility gene, we performed Northern-blot analysis on RNA extracted from Wish, HEL and monocytoid cell lines U-937 and THP-1 treated with 1,000 IU/ml of recombinant IFN-gamma. In U-937 and THP-1 cells, IFN-gamma increased the abundance of RB mRNA. In Wish and HEL cells, co-treatment with cycloheximide was required for IFN-gamma to increase the level of RB mRNA. Pre treatment of THP-1 cells with cycloheximide prior to IFN-gamma treatment augmented the effects of IFN-gamma on RB gene expression. The effect of IFN-gamma in THP-1 cells was observed after 3 hr of treatment, being more pronounced after 6 hr and persisting until at least 18 hr, although at a lower level. These results suggest that IFN-gamma regulates the level of RB mRNA by different mechanisms in the different cell types. This cytokine increases the abundance of RB mRNA in monocytoid cell lines, reinforced by prior treatment with cycloheximide. Inhibition of protein synthesis is required in Wish and HEL cell lines before IFN-gamma has an effect on RB gene expression. PMID- 8416208 TI - Biologic properties of a CH2 domain-deleted recombinant immunoglobulin. AB - Monoclonal antibody (MAb) B72.3 reacts with TAG-72, a high-molecular-weight mucin expressed on several types of human carcinoma, and is currently being used in clinical trials for the diagnosis and therapy of human carcinoma. An expression construct containing cDNA encoding an immunoglobulin (Ig) heavy chain, with the variable region of murine MAb B72.3 and a human Ig constant region with a deletion of the CH2 domain, was generated. Immunoglobulin from the transfectoma with the highest expression of the TAG-72 immunoreactive antibody was designated MAb chimeric (c) B72.3 delta CH2. The pharmacokinetics of serum clearance of iodine-labeled MAbs cB72.3 delta CH2 and the intact cB72.3 were compared in athymic mice. By 24 hr, less than 1% of the cB72.3 delta CH2 was left in the plasma, while 36% of the cB72.3 still remained. The T1/2 alpha values of the cB72.3 delta CH2 and cB72.3 MAbs were 1.7 and 2.4 hr, respectively. The T1/2 beta values were 7.8 hr for the domain-deleted cMAb and 48.9 hr for cB72.3. Biodistribution studies in athymic mice bearing LS-174T xenografts showed a reduction in the percentage of injected dose per gram in tumor with 131I-cB72.3 delta CH2; however, the 131I-cB72.3 delta CH2 both localized to tumors faster and cleared from the blood faster than the 125I-cB72.3 MAb. Only trace amounts of the 131I-cB72.3 delta CH2 were detected in normal tissues, including kidney. The faster clearance rate, more rapid tumor targeting and lack of metabolic uptake in normal tissues demonstrated with the iodine-labeled CH2 domain-deleted cMAb may be an advantage for certain clinical protocols. PMID- 8416209 TI - Publication policies. PMID- 8416210 TI - Computational flow and aerosol concentration profiles in lung bifurcations. AB - Flows in lung bifurcations are complicated by geometry, and it is recognized that accurate lung dosimetric models require realistic calculations of the flow and particle deposition patterns. A computational fluid dynamics study of flow and particle concentration has been carried out for a lung bifurcation based on the model developed by Weibel. The predicted flow patterns match well with previously reported experimental data. Secondary flow patterns and locations close to the walls having high particle concentrations are clearly seen. PMID- 8416211 TI - Factors affecting indoor radon concentrations in the United Kingdom. AB - Data collected in a nationwide study on natural radiation exposure in UK dwellings (Wrixon et al. 1988) were re-analyzed to investigate the effects of rock type and various building and lifestyle characteristics, taken into account simultaneously, on indoor radon concentrations. A multiplicative model which takes into consideration the outdoor radon concentration is used. Indoor radon concentrations were found to be influenced by type of rock underlying the dwelling, double glazing, house type, floor level of rooms in which measurements were taken, window opening habits in the main bedroom, building materials used in the construction of the walls, floor type, and draught proofing. However, these eight factors together account for only 22% of the variation between dwellings. Estimates of the size of the effect associated with each factor are given. PMID- 8416212 TI - Metabolically consistent breathing rates for use in dose assessments. AB - Assessments of doses resulting from exposures to airborne gases and particles are based almost exclusively on inhalation rates that are inconsistent with the quantities of oxygen needed to metabolize dietary intakes of fats, carbohydrates, and protein. This inconsistency leads to erroneous estimates of inhalation exposures and can distort the relative importance of inhalation and ingestion based exposures to environmental contaminants that are present in foods, air, and water. As a means of dealing with this problem, a new methodology for estimating breathing rates is presented that is based on the oxygen uptake associated with energy expenditures and a ventilatory equivalent that relates minute volume to oxygen uptake. Three alternative energy-based approaches for estimating daily inhalation rates are examined: (1) average daily intakes of food energy from dietary surveys, adjusted for under reporting of foods; (2) average daily energy expenditure calculated from ratios of total daily expenditure to basal metabolism; and (3) daily energy expenditures determined from a time-activity survey. Under the first two approaches, inhalation rates for adult females in different age cohorts ranged from 9.7 to 11 m3 d-1, whereas for adult males the range was 13 to 17 m3 d-1. Inhalation rates for adults determined from activity patterns were higher (i.e., 13 to 18 m3 d-1), however, those rates were shown to be quite sensitive to the energy expenditures used to represent light and sedentary activities. In contrast to the above estimates, the ICRP 23 reference values for adult females and males are 21 and 23 m3 d-1 (Snyder et al. 1975). Finally, the paper provides a technique for determining the short-term breathing rates of individuals based on their basal metabolic rate and level of physical activity. PMID- 8416213 TI - Vertical transport of radiocesium in surface soils: model implementation and dose rate computation. AB - A mathematical model on the Radionuclide Behaviour in Soil for the study of the migration of radionuclides in undisturbed soil profile has been developed. The model has been calibrated using 134Cs and 137Cs concentrations along soil samples gathered in a natural grassland and beech wood, respectively, located in the Friuli-Venezia Giulia region (northeastern part of Italy). In these sampling sites, the external exposure due to 134Cs and 137Cs distribution along the soil profile has been also assessed. PMID- 8416214 TI - Bone cancer occurrence among beagles given 239Pu as young adults. AB - The occurrence of skeletal malignancies has been documented among 234 young adult beagles given single intravenous injections of monomeric 239Pu citrate. Occurrence has also been documented among 132 comparable control group animals surviving the minimum latent time period of 2.79 y for radiation-induced bone cancer, who were maintained for lifespan observation. Injected amounts ranged from about 0.02-106 kBq kg-1 body mass with factors of 2 or 3 between dose levels. There were 84 radiographically apparent bone tumors in 76 plutonium injected dogs and one tumor in a control group dog. Most of these were osteosarcomas except for seven chondrosarcomas, one liposarcoma, and one plasma cell myeloma of bone. The relationship between percent of dogs at any dose level with bone malignancy and average skeletal dose at the presumed time of tumor initiation of 1 y before death appeared to be linear below about 1.3 Gy average skeletal dose. The observed data can be approximated by the expression A = 0.76 + 75 D, where A = percent of dogs with bone cancer at any dose level, D = average skeletal dose in Gy (for doses up to 1.3 Gy) at tumor initiation, and 0.76 represents the percent tumor response in the control animals not given plutonium. Similar analysis of our corresponding data for beagles given 226Ra, excluding the two highest dose levels (approximately 100% occurrence), yielded the expression A = 0.76 + 4.7 D, where D = the average skeletal dose in Gy (for doses up to 20 Gy) at 1 y before death. The ratio of coefficients indicates the effectiveness for bone cancer induction of 239Pu relative to 226Ra, or [(75 +/- 22.5)(4.7 +/- 0.47) 1] = 16 +/- 5 for a single, brief intake of either nuclide into blood. PMID- 8416215 TI - 129I and 137Cs fission products in thyroids of animals, 1984-1991. AB - Iodine is intensely concentrated in the thyroid of animals, while 137Cs is not. In this study, 129I and 137Cs concentrations were determined in animal thyroids from selected areas during 1984-1991. The thyroids were from deer killed within the Savannah River Site (SRS) in South Carolina; from the environs of Oak Ridge (OR), Tennessee; West Tennessee; and Florida. Thyroids from sheep slaughtered routinely in Birmingham, England (UK), were also tested. The glands were analyzed by x-ray spectroscopy using a high-purity germanium well detector. 129I concentrations of 1 to 102 Bq 129I (g thyroid)-1 were found in 6.8% of deer thyroids from SRS. Eighty-nine percent of the thyroids from SRS and 38% of those from OR contained 129I concentrations of 0.01 to 1.0 Bq 129I g-1. No thyroids from West Tennessee or Florida had more than 4 mBq 129I g-1. Cesium was found to be distributed differently; 38% of the thyroids from SRS contained 0.1 to 0.65 Bq 137Cs g-1, and the highest concentrations appeared periodically, during November and December. Ten to 100 mBq of 137Cs were found in 60% of thyroids from SRS, 79% of those from Florida, 52% of those from OR, and 22% of those from West Tennessee. In thyroids from SRS and Florida, most of the 137Cs has been from worldwide fallout on sandy soil which permits 137Cs-rich vegetation; if a fraction of the 137Cs is from other sources, it has not been defined. The unique finding of this report is the high incidence and persistence of > 0.01 Bq 129I (g thyroid)-1 in deer from OR and SRS. 129I is a marker for fission products, but it is not a radiological hazard because of its very slow radioactive decay and its continual dilution by additions of nonradioactive iodine to the environment. PMID- 8416216 TI - The effect of the static magnetic field on the response of radiation survey instruments. AB - The effects on the response of eight radiation survey instruments in static magnetic fields up to 0.03 T (300 G) have been investigated. The instruments studied are the Xetex 303B Pacer, the Bicron Micro Rem survey meter, the Victoreen 450p survey meter, the Victoreen 440 survey meter, the SLAC orange meter, the Keithley 36150 survey meter, the Anderson-Braun neutron remmeter, and the Victoreen 488 neutron survey meter. The results show that the effect may depend on several factors such as instrument design, alignment of the instrument axis with the magnetic flux lines, whether the instrument is stationary or moving relative to the magnetic field, the direction of the movement relative to the magnetic flux lines, and the magnetic field intensity. Also presented are results of work to enhance magnetic shielding of some of the instruments. PMID- 8416217 TI - Determining the specific alpha activity of thick sources using a large-area zinc sulfide detector. AB - A simple method using a large-area zinc sulfide detector to determine the total specific alpha activity of thick sources is presented. A previous paper shows how the linear absorption properties of weightless alpha sources can be applied to thick sources placed in direct contact with a varying thickness of window material. A quadratic relationship between the detector response and absorber thickness was derived for sources whose thickness exceeds the range of the alpha particle. The coefficient of the linear term in the quadratic expression is used to calculate the total specific alpha activity of a source in contact with the window of the detector. This relationship is tested by obtaining alpha absorption data from solid sources of known specific alpha activity, fitting the data to the theoretical relationship and comparing the results to the known activities. PMID- 8416218 TI - 137Cs concentrations in lichens before and after the Chernobyl accident. AB - 137Cs activities were measured in a variety of epigeic and epiphytic lichens in Austria before and after contamination by the Chernobyl fallout. For comparison, the activity of the naturally occurring 40K was also determined in each lichen sample. The high 137Cs activities found after Chernobyl suggest that lichens are suitable and inexpensive biological detectors of the fallout pattern. PMID- 8416219 TI - Evaluation of natural radiation in houses built with black schist. AB - Natural radiation in houses built with black schist slabs located at an altitude of 1,000 m in the mountainous southern part of Taiwan were investigated by studying the naturally occurring radionuclides present in the black schist. Both indoor and outdoor radon concentrations were monitored. The cosmic-ray contribution to the dose received by the inhabitants was also estimated. Gamma ray spectroscopy was performed for radionuclide analyses. In situ measurements were carried out using a survey meter coupled to a sodium iodide detector. Cellulose nitrate films, ZnS (Ag) scintillation cells, and alpha spectroscopy were used to study radon and radon daughters. Radiation doses due to all natural sources were calculated and compared with that incurred in common concrete dwellings at lower altitudes. PMID- 8416220 TI - The air-kerma rate constant of 192Ir. AB - The air-kerma rate constant gamma delta (and its precursors), as one of the basic radiation characteristics of 192Ir, was determined by many authors. Analysis of accessible data on this quantity led us to the conclusion that published data strongly disagree. That is the reason we calculated this quantity on the basis of our and many other authors' gamma-ray spectral data and the latest data for mass energy-transfer coefficients for air. In this way, a value was obtained for gamma delta of 30.0 +/- 0.9 a Gy m2 s-1 Bq-1 for an unshielded 192Ir source and 27.8 +/ 0.9 a Gy m2s -1Bq-1 for a standard packaged radioactive source taking into account attenuation of gamma rays in the platinum source wall. PMID- 8416221 TI - Impacts of atmospherically deposited radionuclides on drinking water supply after a nuclear power plant accident. AB - This paper analyzes the various possible situations in which a drinking water supply in China may be contaminated by the deposition of airborne radioactive effluent after reactor accidents occur. A model was utilized to calculate the impact of ingestion of contaminated drinking water from a reservoir after a design basic accident of a 900-MWe pressurized water reactor power station. It showed that the maximum individual effective dose caused by a contaminated drinking water supply was at most 0.02% of the total individual effective dose from a reactor design basic accident. This factor was not considered when evaluating the consequence of a reactor design basic accident since, under several unfavorable conditions, the contamination of a drinking water supply from airborne radioactive effluent after a reactor design basic accident is not a major contributor to the impact on individual effective dose. It may be necessary to consider surface water contamination by deposition of airborne radioactive effluent for an emergency plan after a serious accident. PMID- 8416222 TI - Liquid scintillation sample analysis in microcentrifuge tubes. AB - Local regulations prohibiting drain disposal of "biodegradable" liquid scintillation cocktails prompted investigation of volume reduction for these materials. Microcentrifuge tubes were used with aqueous and filter media samples of 3H, 14C, 32P, and 125I. Backgrounds, counting efficiencies, figures of merit, and spectral distributions obtained for microcentrifuge tubes compared favorably to conventional vials. Differences in 32P spectra for solid support samples appeared related to filter material and sample volume. Decreases in sample costs and waste volume and disposal costs were approximately 50-75%. PMID- 8416223 TI - Thoralf M. Sundt, Jr., M.D., 1930-1992. PMID- 8416224 TI - Colloid cyst of the third ventricle. A comparative immunohistochemical study of neuraxis cysts and choroid plexus epithelium. AB - In an effort to shed light upon the nature of the colloid cyst, the immunohistochemical properties of 21 examples of this lesion were compared with those of other neuraxial cysts and choroid plexus epithelium. The neuraxial cysts included the following: eight Rathke's cleft cysts, 25 pituitaries containing follicular cysts of the pars intermedia, and four enterogenous cysts. Fifteen examples of normal choroid plexus and 12 choroid plexus papillomas were studied as well. These lesions were examined for localization of the following antigens: cytokeratins, epithelial membrane antigen, secretory component, carcinoembryonic antigen, prealbumin, vimentin, glial fibrillary acidic protein (GFAP), S-100 protein, neuron-specific enolase, 68-kD neurofilament protein, chromogranin, serotonin, and lysozyme, and with Leu-7 monoclonal antibodies. Five colloid cysts were immunostained with monoclonal antibodies that were specific for Clara-cell antigens and surfactant, respectively. Sugar moieties were localized using Ulex europaeus I, and Ricinus communis agglutinin I lectins. All Rathke's cleft cysts and follicular cysts of the pars intermedia as well as three selected colloid cysts were examined for pituitary hormones. The epithelial cells of colloid and enterogenous cysts, as well as those lining follicular and Rathke's cleft cyst, showed uniformly strong reactivity for cytokeratins, epithelial membrane antigen, secretory component, and vimentin, and bound Ulex europaeus lectin. Occasional cells in colloid cysts were positive for Clara cell-specific antigens. Reaction for carcinoembryonic antigen was present on the apical surface of scattered cells of colloid, follicular, and Rathke's cleft cysts. Many cells of follicles in the pars intermedia as well as individual cells of five Rathke's cleft cysts were also immunoreactive for chromogranin, S-100 protein, GFAP, and pituitary hormones. Colloid and enterogenous cysts were negative for prealbumin, S-100 protein, GFAP, and neuron-specific enolase; in all but a few instances, they failed to bind Ricinus communis agglutinin. In contrast, normal choroid plexus and choroid plexus papillomas were positive for prealbumin, S-100 protein, neuron specific enolase, cytokeratin, vimentin, and Ricinus communis agglutinin receptors; they lacked Ulex europaeus lectin, 56/66-kD cytokeratins, and epithelial membrane antigen. Unlike normal choroid plexus, choroid plexus papillomas were often GFAP-positive. All tissues studied were nonreactive for lysosome, serotonin, and neurofilament, and with Leu-7 antibodies. This study indicates that the immunophenotype of epithelium lining colloid cysts is similar to that of other cysts showing endodermal or ectodermal differentiation and to respiratory tract mucosa. Epithelium of colloid cysts is immunohistochemically different from that of normal or neoplastic choroid plexus. These findings indicate an endodermal rather than neuroepithelial nature for colloid cysts. PMID- 8416225 TI - Effect of systemic hypotension on cerebral energy metabolism during chronic cerebral vasospasm in primates. AB - The influence of systemic hypotension on cerebral blood flow (CBF) and energy metabolism during chronic cerebral vasospasm after subarachnoid hemorrhage was studied in 15 monkeys. Changes in the phosphorus spectrum, as demonstrated by in vivo phosphorus-31 (31P) magnetic resonance (MR) spectroscopy, or in regional CBF were measured in the parietal cortex during graded hypotension. Sequential changes in the phosphorus spectrum were observed during moderate hypotension in the animals 7 days after the introduction of an autologous blood clot around the right middle cerebral artery (MCA). Angiograms revealed a reduction in vessel caliber by approximately 50% in the right MCA. The mean CBF in the spasm side decreased in parallel with a decrease in the mean arterial blood pressure (MABP) from 120 to 40 mm Hg, indicating the abolition of autoregulation. There were no significant differences in the mean percentage totals of inorganic phosphate (Pi), phosphocreatine (PCr), adenosine triphosphate (ATP), and pH between the hemispheres at baseline MABP before hypotension. The values of PCr, ATP, and pH decreased significantly (p < 0.05) and Pi increased significantly (p < 0.05) at an MABP of less than 60 mm Hg in the involved hemisphere. The ratio of PCr:Pi decreased in parallel with a decrease in MABP. The ATP showed a stepwise decrease during moderate hypotension (MABP 60 mm Hg) and was reduced significantly 20 minutes after the beginning of hypotension (p < 0.05). The results indicate that, during chronic vasospasm, changes in cerebral energy metabolism are coupled with changes in CBF in the state of impaired autoregulation. There exists a critical level for ischemia below which high-energy phosphorus metabolites become markedly depleted. It is suggested that 31P MR spectroscopy may be useful to evaluate the ischemic vulnerability of brain tissue in order to prevent delayed neurological deficit during cerebral vasospasm. PMID- 8416226 TI - Seizure control after surgery on cerebral arteriovenous malformations. AB - Prediction of seizure control after surgery on cerebral arteriovenous malformations (AVM's) is currently unavailable. Between 1982 and 1990, 54 patients (30 males, 24 females) with epilepsy caused by a supratentorial cerebral AVM, without prior manifestation of intracranial hemorrhage, were surgically treated. Patients ranged in age from 11 to 59 years at seizure onset and from 13 to 70 years at surgery; the duration of seizure history ranged from several months to 27 years. The AVM's were located in the temporal (17 cases), frontal (15), parietal (10), rolandic (two), and occipital (two) regions; eight were multilobular. All patients underwent preoperative electroencephalography, intraoperative electrocorticography, and total excision of the AVM; additional cortical excision was performed in 25 cases. Remote seizure foci were identified in the ipsilateral mesial temporal structure in 10 patients with AVM's located in the lateral or posterior temporal lobe and in one with an AVM in the anterior frontal region. Two patients required a second operation to remove a remote seizure focus. Among the 54 patients, there were no operative deaths. After surgical treatment, two patients developed hemiparesis, one had contralateral paresthesia of limbs, two suffered partial visual field defects, and five experienced temporary speech disturbances. Postoperative results of seizure control during follow-up study (mean duration 4.8 years) were excellent in 38 patients (70.4%), good in 10 (18.5%), fair in five (9.3%), and poor in one (1.9%). Results appear to correlate with age at seizure onset, duration of seizures, location of lesions, and cortical excision. Excellent results were shown in 18 (60%) of 30 patients whose age at seizure onset was 30 years or less and in 20 (83.3%) of 24 whose age at seizure onset was greater than 30 years. Eighteen (90%) of 20 patients had excellent results when seizure duration was 1 year or less; only 25% of these underwent cortical excision. Twelve (71%) of the 17 temporal AVM's were associated with demonstrable epileptic foci. Secondary epileptogenesis can occur in humans with supratentorial cerebral AVM's; cortical excision in selected patients can improve the outcome of seizure control. Early surgery of a cerebral AVM in young patients presenting with epilepsy is an important consideration. PMID- 8416227 TI - Posttraumatic spinal cord tethering. Case report. AB - A 53-year-old woman with a complete C-7 traumatic quadriplegia developed progressive neurological deterioration, including bulbar symptoms, 3 years after her initial injury. Magnetic resonance imaging showed tethering of the spinal cord at the level of her injury, with marked stretching of the cervical cord associated with medullary and tonsillar herniation. Following transection of the spinal cord, there was some improvement in her condition. Possible etiological factors accounting for this unique presentation are discussed. PMID- 8416228 TI - Intraspinal extradural meningeal cyst demonstrating ball-valve mechanism of formation. Case report. AB - A diverse collection of unverified theories as to the etiology of extradural meningeal cysts have been previously proposed. One case of intraspinal extradural meningeal cyst of the thoracolumbar region is presented in which a ball-valve mechanism involving an idiopathic dural rent and a herniated segment of an underlying dorsal rootlet was suggested by the operative findings. Closure of the dural rent with marsupialization of the meningeal cyst obliterated this extradural lesion. The ball-valve mechanism of formation and other previously proposed theories are discussed. PMID- 8416229 TI - Rapid spontaneous resolution of acute extradural and subdural hematomas. Case report. AB - The rapid spontaneous resolution of two traumatic acute hematomas, one extradural and one subdural, is reported in a 17-year-old young man. The authors believe that this is the first report of simultaneous resolution of both types of hematoma. PMID- 8416230 TI - Cerebral sparganosis diagnosed and treated with stereotactic techniques. Report of two cases. AB - Infestation of the central nervous system with sparganum, the larva of Spirometra mansoni, is rare. Only 27 cases have been reported in the literature; however, the true incidence might be underestimated. Two cases are reported that were definitively diagnosed by stereotactic biopsy techniques. Without a positive histological diagnosis, the first case would have been diagnosed as having a metastatic brain tumor and a course of radiotherapy would probably have been initiated. Differentiation between granuloma of cerebral sparganosis and brain tumors such as meningiomas, gliomas, and metastatic tumors is often difficult before operation. Detailed magnetic resonance imaging might offer some help in such cases. Emphasis is placed on the increasing importance of stereotactic surgery in the diagnosis and treatment of an intracerebral mass lesion, including cerebral sparganosis. PMID- 8416231 TI - Parvalbumin and calbindin D-28K immunoreactivity in central ganglioglioma and dysplastic gangliocytoma of the cerebellum. Report of two cases. AB - Calbindin D-28K and parvalbumin immunocytochemistry were used in the study of central ganglionic cell tumors. Most neurons in the ganglioglioma were immunoreactive to calbindin D-28K, but a few cells were labeled with antibodies against parvalbumin. In contrast, most cells in dysplastic gangliocytoma of the cerebellum were parvalbumin immunoreactive, but fewer reacted with anti-calbindin antibodies. These latter cells had two or three dendrites with claw-shaped terminals and axons with recurrent collateral branches and varicose terminals filled with strings and buttons. These observations suggest that central ganglionic cell tumors, including dysplastic gangliocytoma of the cerebellum, are composed of neurons which, on the basis of their calcium-binding protein content, have particular metabolic and electrophysiological properties. PMID- 8416232 TI - Implantation of intracerebral depth electrodes for monitoring seizures using the Pelorus stereotactic system guided by magnetic resonance imaging. Technical note. AB - The Pelorus stereotactic system guided by magnetic resonance imaging was used to implant intracerebral depth electrodes for monitoring seizure activity. This stereotactic system is frameless and does not require the use of a computer. It is based on the concept of a ball-and-socket type stereotactic arc director and uses the center-of-arc principle to establish a trajectory for electrode placement. The system not only allows the use of the orthogonal approach, but also provides ample working space and flexibility to choose different entry points and trajectory angles. PMID- 8416233 TI - Herbert Olivecrona: founder of Swedish neurosurgery. AB - Herbert Olivecrona (1891-1980) singlehandedly founded Swedish neurosurgery. At the International Congress in Neurology in Bern in August, 1931, Harvey Cushing invited the cream of the world's medical society to a private banquet. Among the 28 specially invited guests was Herbert Olivecrona. At 40 years old, Olivecrona took his seat with pioneers such as Otfrid Foerster, Percival Bailey, Hugh Cairns, Geoffrey Jefferson, and Sir Charles Sherrington. This suggests that Cushing was impressed by the Swedish aristocrat's didactic deeds when he visited the Serafimer Hospital in Stockholm 2 years earlier. During the mid-1920's, the radiologist Erik Lysholm greatly improved the technique of ventriculography and, challenged by Olivecrona, his diagnostic neuroradiology became of superior quality. In the early 1930's, utilizing technical innovations of his own, Lysholm became a master at demonstrating and localizing posterior fossa tumors, which Olivecrona then operated on. Olivecrona's clinic became the mecca to which many scholars, thirsting for more knowledge, went on a pilgrimage. The international reputation of the clinic was founded, not on epoch-making discoveries, but by the resolute and practical application of methods already launched elsewhere and the exemplary organization that Olivecrona had established in collaboration with Lysholm. In spite of hardships and primitive working conditions, the clinic at the Serafimer Hospital gradually developed into the ideal prototype for a modern neurosurgical department. Olivecrona trained many colorful personalities who later were to lay the foundation for neurosurgery in their home countries; these included Wilhelm Tonnis of Germany, Edvard Busch of Denmark, and Aarno Snellman of Finland. Olivecrona was a true pioneer who made major contributions in practically all fields of conventional neurosurgery. PMID- 8416234 TI - Preservation of hearing in acoustic neurinoma surgery. PMID- 8416235 TI - Perfusion pressure and risk of AVM hemorrhage. PMID- 8416236 TI - Gliomatosis cerebri following radiation and chemotherapy. PMID- 8416237 TI - Control of temporal lobe epilepsy following en bloc resection of low-grade tumors. AB - Thirty-one patients with a mean age of 18.9 years (range 3 to 53 years) who underwent temporal lobe surgery for tumor-related epilepsy over a 14-year period are presented. All had suffered chronic drug-resistant temporal lobe seizures (mean age at onset 6.9 years, range 0 to 30 years; mean duration of condition 11.9 years, range 3 to 39 years). Preoperative interictal scalp electroencephalography tracings indicated unilateral localized epileptic foci in 90% of patients, and computerized tomography scans showed abnormalities within the temporal lobe in 87%. All patients underwent en bloc temporal lobectomy. No patient received adjuvant radiotherapy or chemotherapy. Review of the histological material showed dysembryoplastic neuroepithelial tumor in 27 (87%) of the specimens and microscopic evidence of incomplete removal of tumor in 22 (71%). At long-term follow-up evaluation (mean duration 5.8 years, range 1 to 14 years), 81% of patients were completely free of seizures (Engel grade I) and 10% were almost seizure free (Engel grade II) with no deaths reported in either early or late follow-up review. Only one patient in the series failed to benefit from the surgery. Four patients suffered permanent neurological deficit causing a mild disability. Psychological assessment showed no significant fall in verbal or performance intelligent quotient for the group, but a mild memory impairment was evident in 32%. Behavioral and social aspects improved in nearly all (94%) cases. Relief of seizures could not be predicted by intraoperative electrocorticography, and outcome was independent of the completeness of tumor resection. Postoperative electroencephalographic findings identified epileptiform potentials in 65% of patients, which were associated with a worse seizure-control outcome grade. PMID- 8416238 TI - Vagus nerve stimulation for complex partial seizures: surgical technique, safety, and efficacy. AB - Electrical stimulation of the vagus nerve has shown efficacy in controlling seizures in experimental models, and early clinical trials have suggested possible benefit in humans. Eleven patients with complex partial seizures were subjected to implantation of vagus nerve stimulators. Electrode contacts embedded in silicone rubber spirals were placed on the left vagus nerve in the low cervical area. A transcutaneously programmable stimulator module was placed in an infraclavicular subcutaneous pocket and connected to the electrode. One patient required replacement of the system due to electrode fracture. Another patient developed delayed ipsilateral vocal-cord paralysis; the technique was then modified to allow more tolerance for postoperative nerve edema. A third patient showed asymptomatic vocal-cord paresis on immediate postoperative laryngoscopy. Vagus nerve stimulation produces transient vocal-cord dysfunction while the current is on. Nine patients were randomly assigned to receive either high- or low-current stimulation, and seizure frequency was recorded. The high-current stimulation group showed a median reduction in seizure frequency of 27.7% compared to the preimplantation baseline, while the low-current stimulation group showed a median increase of 6.3%. This difference approached statistical significance. The entire population then received maximally tolerable stimulation. The high-current stimulation group showed a further 14.3% reduction, while the low-current stimulation group showed a 25.4% reduction compared to the blinded period. The efficacy of vagus nerve stimulation seemed to depend on stimulus parameters, and a cumulative effect was evident. These results are encouraging, and further study of this modality as an adjunct treatment for epilepsy is warranted. PMID- 8416239 TI - Odontoid fractures in elderly patients. AB - This retrospective analysis describes the clinical characteristics, treatment, and outcome of 19 patients aged 80 years or older with odontoid fractures. The fractures were due to falls in 15 patients (78.9%) and were associated with motor vehicle accidents in four. Type III fractures were seen in three patients and type II fractures in 16. No patient suffered a neurological injury associated with the fracture. Five patients (26.3%) died during hospitalization; factors contributing to their death included prolonged bed rest, associated injuries, and concomitant medical illnesses. The mean follow-up period in the remaining 14 patients was 28.8 months (range 5 to 72 months). Eight patients with a posterior displacement of 5 mm or less were treated with cervical immobilization, three of whom showed a stable non-union of the fracture site at follow-up review. One patient with 10-mm displacement refused operative treatment. Three of the patients without surgical treatment subsequently died from unrelated causes; all remaining patients resumed their routine activity. Five patients with displacement of 5 mm or greater and instability at the fracture site were treated with posterior cervical fusion of C1-2 using wire and autologous iliac bone grafts. In this group, no operative morbidity or mortality occurred and stable constructs developed in all patients; one patient died from an unrelated cause during the follow-up period and the other patients resumed their normal activity. Prolonged bed rest caused respiratory complications in two of six patients who survived initial hospitalization; two of three patients treated with rigid immobilization developed complications that required alternative treatments. PMID- 8416240 TI - The natural history and management of symptomatic and asymptomatic vertebral hemangiomas. AB - Fifty-nine cases of vertebral hemangioma were seen at the Mayo Clinic between 1980 and 1990. Vertebral hemangiomas were discovered incidentally in 35 patients, while pain was the presenting complaint in 13 patients. Five patients presented directly with progressive neurological deficit requiring surgery, and six patients had surgery elsewhere for spinal cord compression and were referred for follow-up evaluation. To better define the natural history of these lesions, a historical review of these patients was conducted; progression of an asymptomatic or painful lesion to neurological symptoms was found in only two cases (mean follow-up period 7.4 years, range 1 to 35 years). New-onset back pain followed by subacute progression (mean time to progression 4.4 months, range 0.25 to 12 months) of a thoracic myelopathy was the most common presentation for patients with neurological deficit. Initially, all 11 patients with spinal cord compression underwent decompressive surgery with full neurological recovery. Recurrent neurological symptoms were observed in three of six patients following subtotal tumor resection and postoperative administration of 1000 cGy or less radiation therapy (mean follow-up period 8.7 years, range 1 to 17 years). No recurrences were noted in four patients who had subtotal excision plus radiotherapy between 2600 and 4500 cGy. One other patient had gross total tumor removal without radiotherapy and has not had a recurrence. Based on these patients and a review of the literature, the authors recommend annual neurological and radiological examinations for patients with hemangiomas associated with pain, especially young females with thoracic lesions in whom spinal cord compression is most likely to develop. Radiation therapy or embolization is an effective therapeutic alternative for patients with severe medically refractory pain. Regular follow-up monitoring for patients with asymptomatic lesions is unnecessary unless pain develops at the appropriate spinal level. It is concluded that management of patients with a progressive neurological deficit should include preoperative angiography and embolization, decompressive surgery with the approach determined by the degree of vertebral involvement and site of spinal cord compression, and postoperative radiation therapy in patients following subtotal tumor removal. Operative management and complications are discussed. PMID- 8416241 TI - Treatment of spasmodic torticollis with intradural selective rhizotomies. AB - To determine the effects of ventral cervical and selective spinal accessory nerve rhizotomy on spasmodic torticollis, 58 patients who had undergone surgery between 1979 and 1987 were reviewed retrospectively. At the time of surgery, each nerve rootlet was electrically stimulated to determine its effect on the nuchal musculature prior to sectioning. Forty-nine patients (85%) had a marked improvement in their condition, with 33 (57%) attaining an excellent result and 16 (28%) noting significant improvement. Patients complained of abnormal head posture, nuchal muscle spasms, and pain prior to surgery. Muscle spasms were completely relieved in 42 patients (72%) and markedly reduced in 10 (17%). Of the 47 patients with preoperative pain, 30 (64%) were free of their pain and eight (17%) noted that the pain was reduced in intensity and frequency. Thirty-four patients (59%) reported that their resting head posture was restored to a neutral position. The likelihood that a patient's head posture returned to normal was inversely proportional to the preoperative duration of the spasmodic torticollis. Twenty-six patients (45%) suffered mild transient difficulty with swallowing solid foods in the immediate postoperative period. In most cases these minor difficulties abated in the months following surgery. PMID- 8416242 TI - Seizure outcome in patients with surgically treated cerebral arteriovenous malformations. AB - A series of 280 cases of cerebral arteriovenous malformations (AVM's) treated surgically between June, 1970, and June, 1989, is reviewed with particular focus on the preoperative seizure history and follow-up seizure status. Follow-up evaluation (mean duration 7.5 years) was achieved in 98% of cases and was accomplished through re-examinations, telephone interviews, and written questionnaires. Overall, 89% of the surviving patients with a follow-up period of greater than 2 years were free of seizures at last examination. Of the 280 patients in this series, 163 had experienced no seizures preoperatively. A recent follow-up study (with a minimum duration of 2 years or to death) was available in 157 of these 163 cases; 21 patients had died. Of the 136 surviving patients, only eight (6%) were having new ongoing seizures. In the 128 (94%) who had remained seizure-free, 73% were receiving no anticonvulsant agents while 27% were taking anticonvulsant prophylaxis. The 2-year minimum follow-up study in 110 of the 117 patients with preoperative seizures revealed that eight (7%) had died. Of the 102 surviving patients, 85 (83%) were seizure-free (with 48% no longer receiving anticonvulsant therapy), while 17 (17%) still suffered intermittent seizures. However, of these 17 patients, 13 reported their seizures to be improved compared to preoperatively; the seizures were the same in two patients and were worse in two patients. An actuarial analysis was conducted comparing the life expectancy of patients following surgery for AVM's with the expected survival of a general white population of the same age and sex in the West Northcentral region of the United States. No statistically significant difference was found. There were seven perioperative deaths (three from cerebral hemorrhage, two from pulmonary emboli, and two from obstruction of venous drainage) and 22 deaths during the follow-up period. Of these 22 deaths, the cause was unknown in four patients, apparently unrelated to the AVM in 13, and directly or indirectly related to the patient's neurological condition prior to surgery or due to surgery performed for resection of the AVM in five. There was a statistically significant relationship between the size and location of the AVM and the clinical presentation. Patients with small AVM's (< 3 cm) were more likely to present with hemorrhage whereas those with large AVM's were more likely to present with seizures.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8416243 TI - Effect of THAM upon outcome in severe head injury: a randomized prospective clinical trial. AB - Although mortality and morbidity rates from head injury have been reduced substantially by improved prehospital interventions, intensive care, and aggressive management of intracranial pressure (ICP), successful treatment of the primary brain injury has been elusive. In experimental models, tromethamine (THAM) has been effective in treating head injury; this drug acts by entering the cerebrospinal fluid compartment, reducing cerebral acidosis and ICP, and reversing the adverse effects of prophylactic hyperventilation on early recovery. In this randomized prospective clinical trial, THAM was studied to determine if it had beneficial effects in the early management of severe head injuries and if the adverse effects of hyperventilation could be prevented. A total of 149 patients with severe head injury (Glasgow Coma Scale scores of < or = 8) were randomly assigned to either a control or a THAM group. Both groups of patients matched in terms of clinical parameters, including age, sex, number of surgical mass lesions, number in each Glasgow Coma Scale stratum, and first ICP measurement. All patients were treated by a standard management protocol, intubated, mechanically ventilated, and maintained in the pCO2 range of 32 to 35 mm Hg for 5 days. Tromethamine was administered as a 0.3-M solution in an initial loading dose (body weight x blood acidity deficit, average 4.27 cc/kg/hr) given over 2 hours, followed by a constant infusion of 1 ml/kg/hr for 5 days. Outcome was measured at 3, 6, and 12 months postinjury. Although analysis indicated no significant difference in outcome between these two groups at 3 months, 6 months, and 1 year, there was a difference regarding ICP. The time that ICP was above 20 mm Hg in the first 48 hours postinjury was less in patients treated with THAM (p < 0.05). Also, the number of patients requiring barbiturate coma was significantly less in the THAM group (5.48% vs. 18.4%, p < 0.05). The authors conclude that THAM ameliorates the deleterious effect of prolonged hyperventilation, may be beneficial in ICP control, and warrants further study as to the dosage and timing of administration. PMID- 8416244 TI - The effect of embolization with N-butyl cyanoacrylate prior to surgical resection of cerebral arteriovenous malformations. AB - Endovascular therapy of cerebral arteriovenous malformations (AVM's) is an accepted adjunct to surgical therapy. However, the literature has not characterized the benefits or the liabilities of preoperative embolization. This series compares two groups of patients who underwent surgical resection of a cerebral AVM; one group (20 patients) received preoperative transfemoral selective embolization with N-butyl cyanoacrylate (NBCA) and the other group (13 patients) did not. In the group with preoperative embolization, the AVM's were larger (3.9 vs. 2.3 cm) and of a higher Spetzler-Martin grade (3.2 vs. 2.5) as compared to the nonembolized group. The NBCA embolization facilitated surgical resection. Arteries supplying the vascular malformation were readily distinguished from those supplying the normal brain parenchyma. Embolized vessels were compressible and easily cut with microscissors. No bleeding occurred from transected vessels. Operative time and intraoperative blood loss for the two groups were not statistically different, despite the significant differences in lesion size and grade. Endovascular complications included immediate and delayed hemorrhage (15%) and transient ischemia (5%); there were no embolization-related deaths. Postoperative complications for both groups included hemorrhage (15%), residual AVM (6%), and cerebrospinal fluid leak (3%); the mortality rate was 3%. There was no statistically significant difference in surgical complications between the embolized and nonembolized groups. Most patients (91%) in both groups had an excellent or good late neurological outcome, with no significant difference between the groups. This study concludes that preoperative NBCA embolization of AVM's makes lesions of larger size and higher grade the surgical equivalent of lesions of smaller size and lower grade by reducing operative time and intraoperative blood loss, with no statistically significant difference in surgical complications or long-term neurological outcome. PMID- 8416245 TI - Multiloculated hydrocephalus: craniotomy and fenestration of intraventricular septations. AB - Ten pediatric patients with multiloculated hydrocephalus caused by neonatal meningitis, ventriculitis, or intraventricular hemorrhage were surgically treated over a 14-year period (January 1, 1976, to December 31, 1990). Six patients underwent craniotomy and transcallosal fenestration of intraventricular septations followed by placement of a shunt, while the other four were treated by shunting procedures alone. Craniotomy resulted in reduction of the shunt revision rate from a median of 2.75 per year prior to fenestration to 0.25 per year following fenestration, with median observation periods of 44.5 and 27 months, respectively. This was compared to a median revision rate of 0.55 per year for patients treated with shunting procedures alone. There were no deaths in either group. Although no surgical complications were encountered, one patient did require a second fenestration procedure. The important aspects of multiloculated hydrocephalus, including pathophysiology, radiographic correlates, and treatment options, are discussed. The goal of treatment is to eliminate the need for multiple shunt revisions, minimizing the accompanying morbidity and expense. It is concluded that craniotomy and transcallosal fenestration of intraventricular septations is a successful treatment of multiloculated hydrocephalus. PMID- 8416246 TI - Neurosurgical treatment of hypothalamic hamartomas causing precocious puberty. AB - Five children, three girls and two boys, were treated for precocious puberty secondary to hypothalamic hamartoma by resection of the hamartoma. The patients' ages at onset of pubertal development ranged from 6 to 19 months. The hamartomas ranged in size from 6 to 10 mm; four were pedunculated, one was sessile, and all were located below the tuber cinereum. The hamartomas were excised via a right subtemporal approach, with transection at the inferior surface of the hypothalamus; two were adherent posteriorly to the basilar artery and brain stem, and the adhesions were divided. Postoperatively, three children exhibited a transient oculomotor paresis and one other child required eye-muscle surgery. The symptoms and signs of precocious puberty completely regressed postoperatively in all patients. Preoperative hormone assays of testosterone, luteinizing hormone, and follicle-stimulating hormone were within the pubertal range in all five children; postoperative assays fell to prepubertal levels. The children have been followed for 0.5 to 10.5 years (mean 5.0 years) postoperatively, without evidence of recurrence of precocious puberty. One child has begun spontaneous puberty at a normal age. It is concluded that complete resection of hypothalamic hamartomas causing precocious puberty is curative. PMID- 8416247 TI - Metabolism in the globus pallidus after fetal implants in rats with nigral lesions. AB - The effect of fetal mesencephalic implants on glucose utilization in selected brain structures and on apomorphine-induced rotational behavior was measured in rats with a unilateral lesion of the substantia nigra. Ipsilateral, but not contralateral, implants decreased the rotational behavior induced by apomorphine. In addition, the nigral lesion decreased glucose utilization in the dorso- and ventrolateral quadrants of the striatum and in the entopeduncular nucleus but increased glucose utilization in the ipsilateral globus pallidus and lateral habenula. The increased metabolism in the globus pallidus was attenuated by ipsilateral, but not contralateral, mesencephalic implants which also decreased glucose utilization in the dorsomedial caudate nucleus. These results indicate that the ability of an ipsilateral mesencephalic graft to ameliorate the motor behavior in rats with nigral lesions is associated with changes in the functional activity of the ipsilateral globus pallidus. PMID- 8416248 TI - Comparison of regeneration across nerve allografts with temporary or continuous cyclosporin A immunosuppression. AB - The efficacy of short-term immunosuppression in a nerve allograft model was examined by comparing regeneration across peripheral nerve allografts with either temporary (12 weeks) or continuous (30 weeks) cyclosporin A treatment. One hundred fifty Lewis rats received 2-cm nerve grafts from allogeneic ACI or syngeneic Lewis rat donors and were allocated to the following groups: allogeneic grafts and continuous cyclosporin A, with 18 weeks (20 rats) or 30 weeks (20 rats) of survival after graft placement; allogeneic grafts and temporary cyclosporin A, with 12 weeks (10 rats), 18 weeks (20 rats), or 30 weeks (20 rats) of survival; and control rats with allogeneic and syngeneic grafts, no cyclosporin A, with 12, 18, or 30 weeks (10 rats each) of survival. Functional regeneration across the nerve grafts was serially assessed with walking-track analysis. Endpoint evaluations included electrophysiological, histological, and morphometric studies. Both walking-track and electrophysiological function reached a plateau at a significantly worse level in nonimmunosuppressed allograft recipients than in syngeneic or treated allograft recipients. The group with temporary therapy experienced significant worsening in both motor and electrophysiological function at Week 18, 6 weeks after cyclosporin A withdrawal, compared to the group with continuous treatment. At Week 30, motor and electrophysiological function in the temporary-treatment group recovered to levels similar to those of the syngeneic and continuous cyclosporin A groups. Histological assessment of the graft segments from the temporary cyclosporin A group at 18 weeks showed evidence of rejection, with mononuclear cell infiltration and demyelination; morphometric evaluation demonstrated significantly decreased numbers of nerve fibers in the distal host segment. These histological and morphometric changes were no longer present in the nerves from the temporarily immunosuppressed rats at Week 30. Withdrawal of immunosuppression after successful regeneration through nerve allografts results in short-term graft rejection. Eventual restoration of graft histological and function parameters is comparable to continuously immunosuppressed rats. Temporary immunosuppression of nerve allograft recipients is feasible. PMID- 8416249 TI - California registries expand SEER's minority coverage. PMID- 8416250 TI - Patients' quality of life under study in Europe. PMID- 8416251 TI - RAC defers gene therapy approval. PMID- 8416252 TI - Breast cancer after treatment of Hodgkin's disease. AB - BACKGROUND: Most studies of survivors of Hodgkin's disease have shown a low risk for subsequent breast cancer, even though much lower doses of radiation than those used for Hodgkin's disease have been shown to induce breast cancer in other settings. PURPOSE: This study quantifies the risk of breast cancer following Hodgkin's disease treatment according to age at treatment and type of treatment. METHODS: To evaluate the risk of breast cancer from irradiation, we reviewed records of 885 women treated for Hodgkin's disease between 1961 and 1990 (mean follow-up, 10 years). Risks for breast cancer incidence and mortality were calculated by comparison with expected rates for a general female population matched by age and race. RESULTS: Twenty-five patients have developed invasive breast cancer, yielding a relative risk (RR) of 4.1 (95% confidence interval [CI] = 2.5-5.7). An additional patient developed multifocal carcinoma in situ. Age at irradiation strongly influenced risk: RR was 136 for women treated before 15 years of age (95% CI = 34-371). RR declined with age at irradiation (P for trend < .0001), but the elevation remained statistically significant for subjects less than 30 years old at the time of irradiation (for those 15-24, RR = 19 [95% CI = 10.3-32]; for those 24-29, RR = 7 [95% CI = 3.2-14.4]). In women above 30 years of age, the risk was not elevated (RR = 0.7; 95% CI = 0.2-1.8). Risk of breast cancer increased significantly with time since treatment (P for trend < .0001). The RR was 2.0 (95% CI = 1.0-3.5) with follow-up under 15 years and 13.6 (95% CI = 7.9-18.2) with follow-up equal to or exceeding 15 years. The addition of mechlorethamine, vincristine, procarbazine, and prednisone chemotherapy to irradiation increased the risk within the first 15 years. Most breast cancers (22 of 26) arose within or at the margin of the radiation field and were infiltrating ductal carcinomas. Stage distribution and outcome suggest that the increased incidence was not solely attributable to vigilant screening. RR of death from breast cancer was 5.1 (95% CI = 2.2-10.0). CONCLUSIONS: Women treated for Hodgkin's disease with radiation before 30 years of age are at markedly increased risk for breast cancer, with risk increasing dramatically more than 15 years after therapy. IMPLICATIONS: The high RR for development of breast cancer in women exposed to therapeutic radiation under 30 years of age raises important issues about optimal treatment strategies for patients with Hodgkin's disease, breast cancer, and other cancers. PMID- 8416253 TI - Treatment failure and dietary habits in women with breast cancer. AB - BACKGROUND: Epidemiological and experimental evidence suggests that breast cancer risk can be reduced by dietary measures. Study of the relationships between dietary habits and prognosis in patients with breast cancer is essential to the design of diet intervention trials. PURPOSE: Our purpose was to determine whether dietary habits are associated with disease-free survival in patients with breast cancer who have undergone treatment. METHODS: We interviewed 240 women about their dietary histories. These women were 50-65 years old and had pathological stage I-II breast cancer with subsequent follow-up for 4 years; 209 of these women were postmenopausal. Differences in dietary variables between groups of patients were analyzed with bivariate and multivariate statistical methods. RESULTS: Cancers were classified as estrogen receptor (ER) rich (> or = 0.10 fmol/micrograms of DNA) in 149 patients and as ER poor (< 0.10 fmol/micrograms of DNA) in 71 patients. Fifty-two patients had treatment failure during follow-up. The 30 patients with ER-rich tumors who had treatment failure reported higher intakes of total fat, saturated fatty acids, and polyunsaturated fatty acids than did the 119 patients with ER-rich tumors who did not have treatment failure. The multiple-odds ratio (OR) for treatment failure in these women was 1.08 for each 1% increment in percentage of total energy (E%) from total fat. For treatment failure within the first 2 years, the OR was 1.19 for each 1-mg increase in vitamin E intake per 10 megajoules of energy. In women with treatment failure 2-4 years after diagnosis, ORs were 1.13 and 1.23 for each E% increment in total fat or saturated fatty acids, respectively. No association between dietary habits and treatment failure was found for women with ER-poor cancers. There was a tendency to a dose-response relationship (in quartiles) between intake of saturated fatty acids and disease-free survival, but the observed differences were not statistically significant. CONCLUSIONS: Dietary habits at the time of diagnosis may affect prognosis, at least for patients with ER-rich breast cancers. Dietary fat may have an effect on growth or spread of breast cancer, both of which may vary according to type of fat. Total fat and saturated fatty acids were the dietary parameters most strongly associated with risk for treatment failure. IMPLICATIONS: Dietary intervention might serve as an adjuvant treatment to improve breast cancer prognosis. PMID- 8416254 TI - Second cancer after Hodgkin's disease--the price of success? PMID- 8416255 TI - Prolonged continuous infusion of fluorouracil with weekly bolus leucovorin: a phase II study in patients with disseminated colorectal cancer. AB - BACKGROUND: Biochemical modulation of bolus fluorouracil (5-FU) by addition of leucovorin to the treatment regimen has increased response in patients with disseminated colorectal cancer from fewer than 20% to more than 40%. In view of the short half-life of 5-FU and its cell cycle specificity, it may be that infusion rather than intravenous bolus injection would increase efficacy. Furthermore, the advent of safer indwelling intravenous catheters and pump technology, allowing home and ambulatory treatment, has made protracted infusion clinically feasible. To examine these questions, we conducted a phase I trial using protracted infusion of 5-FU by indwelling catheter and pump, with leucovorin given by bolus injection, and reported 40% partial response. PURPOSE: We have now initiated a phase II study of 5-FU given by prolonged continuous infusion with weekly bolus injections of leucovorin in previously untreated patients with measurable, disseminated colorectal cancer. METHODS: Forty-one patients were treated. The regimen consisted of treatment for 4 weeks with 5-FU at a dose of 200 mg/m2 daily as a continuous infusion by indwelling intravenous catheter and pump, followed by a 2-week rest and then by monthly cycles of 3 weeks of treatment and 1-week rest until disease progression. Leucovorin was given as a bolus injection of 20 mg/m2 at the beginning of each week of treatment with 5-FU. RESULTS: Nineteen (46%) of 41 patients had objective response: Three complete responses and 16 partial responses were seen. Overall, the median duration of response was 8 months. The median duration of survival was 16 months: 18 months for responders and 10 months for nonresponders. In general, toxic effects were mild and consisted primarily of stomatitis and palmar-plantar erythrodysesthesia (hand-foot syndrome). Neither grade 4 toxic effects nor treatment-related deaths were observed. The only serious side effects were catheter thrombosis (three patients) and catheter sepsis (one patient). CONCLUSION: We conclude that this safe regimen is one of the most effective for the treatment of disseminated colorectal cancer. IMPLICATIONS: The regimen should be tested prospectively against other regimens in use for this disease. It is currently included in a phase III study of the Southwest Oncology Group for this purpose. That study will assess quality of life as well as response rates and survival duration. PMID- 8416256 TI - Regression of oral leukoplakia with alpha-tocopherol: a community clinical oncology program chemoprevention study. AB - BACKGROUND: Oral leukoplakia is an important model for developing chemoprevention approaches for lesions in the upper aerodigestive tract. These lesions most often result from exposure to carcinogens such as tobacco and alcohol and may precede development of invasive cancer. The potent antioxidant alpha-tocopherol (vitamin E) has prevented the development of cancers of the oral cavities in animal models. PURPOSE: The objectives of this study were to evaluate the toxicity and efficacy of alpha-tocopherol in patients with oral leukoplakia and to assess the feasibility of performing chemoprevention trials through the network of the Community Clinical Oncology Program (CCOP). METHODS: A single-arm phase II study using the nontoxic agent alpha-tocopherol to treat oral premalignant leukoplakia was performed at seven institutions affiliated with the CCOP through The University of Texas M. D. Anderson Cancer Center. Patients with symptomatic leukoplakia or dysplasia were treated orally with alpha-tocopherol (400 IU) twice daily for 24 weeks. Follow-up was performed at 6, 12, and 24 weeks after the start of treatment to assess toxicity and response, and serum alpha-tocopherol levels were determined at baseline and at 6 and 24 weeks. RESULTS: Of the 43 patients who have completed 24 weeks of treatment, 20 (46%) had clinical responses and nine (21%) had histologic responses. Mean serum alpha-tocopherol levels were 16.1 micrograms/mL at baseline and increased to 34.29 micrograms/mL after 24 weeks of treatment. Patient-recorded drug calendars, as well as serum drug levels, indicated excellent patient compliance; an average of 95% of the prescribed pills were taken. Treatment was extremely well tolerated; no grade 3 or 4 toxic effects were reported. CONCLUSIONS: Administration of alpha-tocopherol resulted in both clinical and histologic responses in premalignant leukoplakia lesions. The study also demonstrated that chemoprevention trials can be performed through the CCOP. The major problems were that a high percentage of patients were not assessable for response, some patients withdrew because expenses were not reimbursable, and there was limited participation within the CCOP network. These problems may reflect difficulties inherent in the implementation of multi institutional chemoprevention trials. IMPLICATIONS: The efficacy of alpha tocopherol alone and in combination with other chemopreventive agents for carcinogenesis in the upper aerodigestive tract should be explored in future trials. PMID- 8416257 TI - Nutrition and breast cancer. PMID- 8416258 TI - Environmental tobacco smoke: the price of scientific uncertainty. PMID- 8416259 TI - Radar guns. PMID- 8416260 TI - Steady tumor growth and the spontaneous disappearance of cancer. PMID- 8416261 TI - Drugs acting on the intracellular signalling system. AB - Cellular response to extracellular messages is a basic process to maintain and to support cell life. Several signalling molecules important as sites of therapeutic drug action are involved in the response. Recent studies on life sciences have elucidated molecular properties of intracellular signalling factors and mechanisms of cascading. Novel drugs acting on signalling molecules and possessing new sites and mechanisms of action have been found. This article summarizes the properties (subtypes, structures, functions) of signalling factors (receptors, ion channels, GTP binding proteins, second messenger-generating enzymes, second messenger-metabolizing enzymes, second messengers protein kinases, protein phosphatases) and lists in Tables A-H drugs that act on signalling molecules and which should find clinical use. PMID- 8416262 TI - Growth factor second messenger systems: oncogenes and the heterotrimeric GTP binding protein connection. AB - We feel that there is now compelling evidence that the GTP-binding proteins play more than just a coordinating role in the actions of both tyrosine kinase and nontyrosine kinase receptor signal transduction. These similarities appear to represent just a small component of the convergence in the signaling pathways for structurally dissimilar receptor subsets. Future years will see further understanding of the intricacies of these G-protein-proto-oncogene interactions, and the extension into the potential role in growth factor action played by the expanding number of known members of this G-protein family. PMID- 8416263 TI - Ivermectin, an antiparasitic agent. PMID- 8416264 TI - Neuropharmacology of potassium ion channels. PMID- 8416265 TI - The diagnosis of thoracic aortic dissection by noninvasive imaging procedures. AB - BACKGROUND AND METHODS: This study was designed to assess the safety and reliability of new noninvasive imaging methods as compared with aortography in the diagnosis of dissection of the thoracic aorta. One hundred ten patients with clinically suspected aortic dissection followed a diagnostic protocol that included transthoracic and transesophageal color-flow Doppler echocardiography (TTE and TEE), contrast-enhanced x-ray computed tomography (CT), and magnetic resonance imaging (MRI). Imaging results were compared in a blinded fashion and validated independently against intraoperative findings in 62 patients, autopsy findings in 7, and the results of contrast angiography in 64. RESULTS: The sensitivities of MRI, TEE and x-ray CT for detecting dissection were similar, at 98.3, 97.7, and 98.3 percent, respectively; TTE had a sensitivity of only 59.3 percent (P < 0.005). The specificities of both TTE (83.0 percent) and TEE (76.9 percent) were lower than those of x-ray CT (87.1 percent) and MRI (97.8 percent; P < 0.05), mainly as a result of false positive findings in the ascending aorta. MRI and x-ray CT were more sensitive than TTE in detecting the formation of thrombus in the entire thoracic aorta (P < 0.05), but were not superior to TEE in this regard. CT was not effective in detecting an entry site or aortic regurgitation, but MRI and TEE accurately identified both. Two patients died during or soon after CT and TEE, and three died between retrograde angiography and surgery. CONCLUSIONS: A noninvasive diagnostic strategy using MRI in all hemodynamically stable patients and TEE in patients who are too unstable to be moved should be considered the optimal approach to detecting dissection of the thoracic aorta. Comprehensive and detailed evaluation can thus be reduced to a single noninvasive diagnostic test in the investigation of suspected dissection of the thoracic aorta. PMID- 8416266 TI - Association between polymorphism of the glycogen synthase gene and non-insulin dependent diabetes mellitus. AB - BACKGROUND: The storage of glucose as glycogen in skeletal muscle is frequently impaired in patients with non-insulin-dependent diabetes mellitus (NIDDM) and their nondiabetic relatives. Despite an intensive search for candidate genes associated with NIDDM, no data have been available on the gene coding for the key enzyme of this pathway, glycogen synthase. METHODS AND RESULTS: Using a human complementary DNA probe, the restriction enzyme Xbal, and Southern blot analysis, we identified two polymorphic alleles, A1 and A2, in the glycogen synthase gene. The gene was localized to chromosome 19. The A1A2 or A2A2 genotype was found in 30 percent of 107 patients with NIDDM but in only 8 percent of 164 nondiabetic subjects without a family history of NIDDM (P < 0.001). The diabetic patients with the A2 allele had a stronger family history of NIDDM (P = 0.019), a higher prevalence of hypertension (P = 0.008), and a more severe defect in insulin stimulated glucose storage (P = 0.001) than the diabetic patients with the A1 allele. The concentration of the glycogen synthase protein in biopsy specimens of skeletal muscle from the patients with the A2 allele was normal, however, suggesting that expression of the gene was unaltered. The Xbal polymorphism was due to a change of a single base in an intron. CONCLUSIONS: The Xbal polymorphism of the glycogen synthase gene identifies a subgroup of patients with NIDDM characterized by a strong family history of NIDDM, a high prevalence of hypertension, and marked insulin resistance. PMID- 8416267 TI - Comparison of low-dose isotretinoin with beta carotene to prevent oral carcinogenesis. AB - BACKGROUND: High-dose isotretinoin therapy has been determined to be an effective treatment for leukoplakia. However, a high rate of relapses and toxic reactions led us to conduct a trial of a much lower dose of isotretinoin in the hope of maintaining a response and limiting toxicity. METHODS: In the first phase of the study, 70 patients with leukoplakia underwent induction therapy with a high dose of isotretinoin (1.5 mg per kilogram of body weight per day) for three months; in the second phase, patients with responses or stable lesions were randomly assigned to maintenance therapy with either beta carotene (30 mg per day) or a low dose of isotretinoin (0.5 mg per kilogram per day) for nine months. RESULTS: In the first phase, the rate of response to high-dose induction therapy in the 66 patients who could be evaluated was 55 percent (36 patients). The lesions of seven patients progressed, and therefore they did not participate in the second phase of the trial. Of the 59 patients included in the second phase, 33 were assigned to beta carotene therapy and 26 to low-dose isotretinoin therapy; these two groups did not differ significantly in prognostic factors. Of the 53 patients who could be evaluated, 22 in the low-dose isotretinoin group and 13 in the beta carotene group responded to maintenance therapy or continued to have stable lesions (92 percent vs. 45 percent, P < 0.001). In situ carcinoma developed in one patient in each group, and invasive squamous-cell carcinoma in five patients in the beta carotene group. Toxicity was generally mild, though greater in the group given low-dose isotretinoin therapy. CONCLUSIONS: When preceded by high dose induction therapy, low-dose isotretinoin therapy was significantly more active against leukoplakia than beta carotene and was easily tolerated. PMID- 8416268 TI - Acute bacterial meningitis in adults. A review of 493 episodes. AB - BACKGROUND AND METHODS: To characterize acute bacterial meningitis in adults, we reviewed the charts of all persons 16 years of age or older in whom acute bacterial meningitis was diagnosed at Massachusetts General Hospital from 1962 through 1988. We included patients who were admitted after initial treatment at other hospitals. RESULTS: During the 27-year period, 445 adults were treated for 493 episodes of acute bacterial meningitis, of which 197 (40 percent) were nosocomial. Gram-negative bacilli (other than Haemophilus influenzae) caused 33 percent of the nosocomial episodes but only 3 percent of the community-acquired episodes. In the 296 episodes of community-acquired meningitis, the most common pathogens were Streptococcus pneumoniae (37 percent), Neisseria meningitidis (13 percent), and Listeria monocytogenes (10 percent); these organisms accounted for only 8 percent of the nosocomial episodes. Only 19 of the 493 episodes of meningitis (4 percent) were due to H. influenzae. Nine percent of all patients had recurrent meningitis; many had a cerebrospinal fluid leak. Seizures occurred in 23 percent of patients with community-acquired meningitis, and 28 percent had focal central nervous system findings. Risk factors for death among those with single episodes of community-acquired meningitis included older age (> or = 60 years), obtunded mental state on admission, and seizures within the first 24 hours. Among those with single episodes, the in-hospital mortality rate was 25 percent for community-acquired and 35 percent for nosocomial meningitis. The overall case fatality rate was 25 percent and did not vary significantly over the 27 years. CONCLUSIONS: In our large urban hospital, a major proportion of cases of acute bacterial meningitis in adults were nosocomial. Recurrent episodes of meningitis were frequent. The overall mortality rate remained high. PMID- 8416269 TI - Diagnostic imaging in the evaluation of suspected aortic dissection. Old standards and new directions. PMID- 8416270 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 1-1993. A seven-year-old girl with recurrent bouts of sore throat, cough, dyspnea, and fever. PMID- 8416271 TI - Diabetes genes in non-insulin-dependent diabetes mellitus. PMID- 8416272 TI - Biologic modifiers and chemoprevention of cancer of the oral cavity. PMID- 8416274 TI - Kidney-related Munchausen's syndrome and the Red Baron. PMID- 8416273 TI - Kidney-related Munchausen's syndrome and the Red Baron. PMID- 8416275 TI - Kidney-related Munchausen's syndrome and the Red Baron. PMID- 8416276 TI - Kidney-related Munchausen's syndrome and the Red Baron. PMID- 8416277 TI - Kidney-related Munchausen's syndrome and the Red Baron. PMID- 8416278 TI - The prevention and treatment of osteoporosis. PMID- 8416279 TI - The prevention and treatment of osteoporosis. PMID- 8416280 TI - Hair analysis as a marker for fetal exposure to maternal smoking. PMID- 8416281 TI - Ernst Richard Jaffe--a tribute. PMID- 8416282 TI - Band 3 peptides inhibit deoxy S polymerization: viscosity studies. AB - We have previously obtained evidence that N-terminal band 3 peptides inhibited deoxyhemoglobin S (deoxy S) polymerization as determined by equilibrium solubility assays. An N:1-15AA fragment binds to the 2,3-diphosphoglycerate (2,3 DPG) receptor locus of deoxy S with five to seven amino acids (AA) extending internally, while ten to eight AA remained external to deoxy S and inhibited polymerization by steric hindrance. A true mirror-image peptide, corresponding to two N:1-8AA + lysine (K) linked by coupler, binds to the 2,3-DPG loci of two deoxy S molecules, tethering them together to form "binary complexes" incapable of entering the polymer chains. The reduction in the concentration of deoxy S available for extended chain formation decreased polymerization. We now report time:viscosity profiles of the sol-gel transformation of purified solutions of deoxy S with and without peptides and studies of the gel solidity at equilibrium. Samples with peptides had longer lag times than controls of similar deoxy S concentrations. The mirror-image peptide was a more effective inhibitor than the N:1-15AA peptide. When the mirror-image peptide was present in peptide:hemoglobin molar ratios of 0.25-1:1, the increases in lag time were equivalent to decreasing the deoxy S concentrations by 15-25%, comparable to projected major therapeutic effects. Gel solidity, determined by yield temperature, was less in the sample with mirror-image peptide compared to control. These results support the proposed mechanisms of inhibition of deoxy S polymerization by band 3 peptides. PMID- 8416283 TI - Some properties of hemoglobin A2. AB - While no significant physiologic function of hemoglobin A2 (Hb A2), the minor basic component of human hemoglobin, has been recognized, only its oxygen equilibria have been studied in detail. Since hemoglobin A2 and its oxidative denaturation product, hemichrome A2, bind to the red cell membrane, particularly to band 3, to a greater extent than do Hb A or hemichrome A, some of the properties of Hb A2 that might influence hemoglobin-membrane association were examined. Hemoglobin A2 exhibited slightly increased susceptibility to autoxidation to methemoglobin. No differences were noted between methemoglobins A and A2, including the rates of enzymatic reduction and stability of the heme globin linkage. Oxyhemoglobin A2 had a slightly lower solubility in phosphate buffer than did hemoglobin A. While the hemichromes (prepared with phenylhydrazine) of hemoglobins A2 and A had the same optical spectra, the A2 hemichrome exhibited greater stability. It is suggested that the differences in products of oxidative denaturation may provide the basis for functional differences between hemoglobins A2 and A. PMID- 8416284 TI - The use of hemoglobin as a blood substitute. PMID- 8416285 TI - Composition of the hemoglobin S polymer. PMID- 8416286 TI - Evolution of clinical understanding: paroxysmal nocturnal hemoglobinuria as a paradigm. AB - In recounting the history of the development of knowledge about PNH, the roles of the clinician, the basic scientist, and the clinician-investigator are apparent. Without the observations of the clinicians, the problem could not be posed. Without the contributions from basic science (the biochemistry of complement, the biology of glycosyl-phosphatidylinositol anchors, etc), the information necessary to the solution of the problem would not be available. Without the synthesis of the clinician-investigator, the two elements would not be fused to result in knowledge about the disease. PMID- 8416287 TI - Adult and childhood acute lymphocytic leukemia: are they different diseases? AB - Age has long been recognized as an important factor in predicting response to treatment for acute lymphocytic leukemia (ALL). Specifically, the results of treatment of childhood ALL have been far superior to the treatment of what appears to be the same disease in adults. However, from an analysis of the clinical and biological prognostic factors known to be predictive in childhood ALL, there is a striking difference in their distribution in adults with ALL. It appears that there is a special form of ALL seen in children of some populations with a peak incidence of three to seven years. This treatment responsive leukemia appears to be different clinically, biologically, and epidemiologically from adult ALL. PMID- 8416288 TI - Evidence that NADPH-dependent methemoglobin reductase and administered riboflavin protect tissues from oxidative injury. AB - NADPH-dependent methemoglobin reductase, first detected in erythrocytes sixty years ago, has subsequently been purified and characterized as a methylene blue reductase and a flavin reductase. The reductase plays no role in methemoglobin reduction under normal conditions, but its activity serves as the basis for the treatment of methemoglobinemia with methylene blue or flavin. On-going studies demonstrate that this cytosolic protein is also present in liver and that its primary structure distinguishes it from other known proteins. The bovine erythrocyte reductase tightly binds hemes, porphyrins, and fatty acids with resulting loss of activity. Pyrroloquinoline quinone serves as a high-affinity substrate of the reductase, suggesting that this naturally-occurring compound may be a physiological substrate. The ability of the reductase to catalyze the intracellular reduction of administered riboflavin to dihydroriboflavin suggested that this system might be exploited to protect tissues from oxidative damage. This hypothesis was supported by our finding that dihydroriboflavin reacts rapidly with Fe(IV)O and Fe(V)O oxidation states of hemeproteins, states that have been implicated in tissue damage associated with ischemia and reperfusion. Preliminary studies demonstrate that, as predicted, administration of low concentrations of riboflavin protects isolated rabbit heart from reoxygenation injury, rat lung from injury resulting from systemic activation of complement, and rat brain from damage caused by four hours of ischemia. Data from these animal studies suggest that flavin therapy holds promise in protecting tissue from the oxidative injuries of myocardial infarction, acute lung injury, stroke, and a number of other clinical conditions. PMID- 8416289 TI - Agranulocytosis during antibiotic therapy: drug sensitivity or sepsis? AB - Forty-three patients reviewed from the literature and five cases of agranulocytosis during antibiotic therapy studied by the author are presented. Time required to develop agranulocytosis with antibiotics was < 19 days in comparison to > 40 days required with nonantibiotic drugs. In all, agranulocytosis occurred concomitantly with drug treatment and became normal as treatment was discontinued. Retrospective rechallenge studies suggest that agranulocytosis may be dose related. In all cases PMNs were almost completely deleted and marrows were devoid of granulocyte precursors. In contrast, leukopenia secondary to overwhelming sepsis displayed persisting granulocytes in peripheral blood and marrow. While leukagglutinins were not found in nine cited cases, four serums were toxic to test PMNs as measured by suppression of postphagocytosis respiratory burst. Clindamycin directly suppressed development of CFU-G in one sensitive patient but not in 16 normal controls. The hazard of antibiotics in suppressing granulocytopoiesis is emphasized by these observations. PMID- 8416290 TI - Inhibition of the activation of hageman factor (factor XII) by eosinophils and eosinophilic constituents. AB - Several syndromes characterized by striking eosinophilia may be complicated by thrombosis. The experiments described indicate that, paradoxically, eosinophils and certain of their constituents inhibit the activation of Hageman factor (HF, factor XII). In earlier studies, suspensions of mixed types of granulocytes, other nucleated peripheral blood cells, and platelets inhibited activation of Hageman factor by ellagic acid, glass, and sulfatides. After these cells were sedimented by centrifugation, the supernatant fluids were also inhibitory. No attempt had been made earlier to distinguish among different granulocytic species. In the present study, suspensions of eosinophils and the supernatant fluid after eosinophils had been separated by centrifugation inhibited activation of Hageman factor by ellagic acid. The protein concentration of that amount of supernatant fluid that inhibited activation by about half was 16 micrograms/ml, approximately the same as had been described for suspensions of peripheral blood mononuclear cells. Activation of Hageman factor by ellagic acid was also inhibited by certain constituents of eosinophils, including eosinophil peroxidase, eosinophil major basic protein and eosinophil cationic protein. Inhibition was not specific for ellagic acid-induced activation of Hageman factor, as inhibition was also observed with sulfatide-induced activation. Inhibition was presumably related to neutralization of the negative charge of activators of Hageman factor. Thus, bismuth subgallate, a particulate activator of Hageman factor, was no longer effective after it had been exposed to eosinophil cationic protein. The observations reported here raise the question of whether in vivo eosinophils modulate certain of the defense reactions ascribed to Hageman factor. PMID- 8416291 TI - Defining the architecture of the red blood cell membrane: newer biophysical approaches. AB - For many years the red cell membrane has served as an extraordinarily valuable model for membrane structure and function. During the past 2 decades, the biochemical concept of the membrane skeleton was established, and, with the help of electron microscopy, a partial depiction of this structure evolved. Newer biophysical approaches designed to measure distances between various components of membrane skeleton as well as distances between the membrane skeleton and the overlying bilayer should now help to define this structure more realistically. Fluorescence resonance energy transfer, single photon radioluminescence, and total internal reflectance are three biophysical techniques that will enable us to measure such distances over a substantial range, which extends from a few Angstroms to approximately 2 microns. The ability to make such measurements in intact cells and in fully hydrated, undenatured membrane preparations should add a new dimension to our understanding of the structure of the red cell membrane. PMID- 8416292 TI - Hereditary hemolytic disease with increased red blood cell phosphatidylcholine and dehydration: one, two, or many disorders? AB - We have compared characteristics of red cells from patients who were originally diagnosed as having two different disorders, high phosphatidyl choline hemolytic anemia (HPCHA) and hereditary xerocytosis (HX). Both types of cells had reduced intracellular potassium, with attendant cell dehydration and an increase in the relative amount of membrane phosphatidyl choline. Neither these observations nor a review of previous studies of HX and HPCHA revealed any means of distinguishing between the two disorders. Measurements of chloride-dependent potassium transport revealed flux characteristics in both HX and HPCHA red cells that were different from those in simultaneously run control samples. HX and HPCHA red cells did not show the same kinds of deviations from the normal pattern. However, extensive characterization of transport behavior under a variety of controlled conditions will be required to determine whether these differences represent intrinsic differences in chloride-dependent transport properties. It appears likely that HX and HPCHA both represent a spectrum of disorders resulting from a variety of defects that produce the same general pattern of abnormalities in cation content and membrane phospholipid composition. PMID- 8416293 TI - Historical note: methemoglobinemia--long ago and far away. PMID- 8416294 TI - Concise review: pyruvate kinase deficiency: historical perspective and recent progress of molecular genetics. PMID- 8416295 TI - Acute episodic hemolysis in the African black rhinoceros as an analogue of human glucose-6-phosphate dehydrogenase deficiency. AB - Sudden episodes of massive hemolysis have become the most common cause of death among captive black rhinoceroses, and there is evidence that they occur in the wild as well. We have observed radically unique enzyme and metabolite profiles in normal rhinoceros erythrocytes compared to humans and other mammals, including marked deficiencies of intracellular adenosine triphosphate (ATP), catalase, adenosine deaminase, and other enzymes involved in glycolysis, glutathione cycling, and nucleotide metabolism. Minimal concentrations of ATP appear to impair effective acceleration of hexosemonophosphate shunt activity in response to oxidants by restricting substrate generation at the hexokinase step. Antioxidant defenses are further compromised by catalase deficiency, which may be a general characteristic of rhinoceros erythrocytes, perhaps related to the common occurrence of severe mucocutaneous ulcerative disease. It is proposed that erythrocyte ATP deficiency in rhinoceroses may be an evolutionary adaptation conferring selective advantage against common hemic parasites, comparable to the role of human glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in falciparum malaria. PMID- 8416296 TI - Mechanism of red blood cell aging: relationship of cell density and cell age. AB - The human red cell has a life span of 120 days. The mechanism that determines cell removal from the circulation with such precision remains unknown. Most studies of red cell aging have been based on analysis of cells of progressively increasing age separated by density. The relationship between red cell age and density has been recently challenged, and the hypothesis has been put forward that cell death is not the result of a progressive deterioration of essential cell constituents. This theory was based on preliminary observations in transient erythroblastopenia of childhood, which could not later be confirmed. When the relationship between cell aging and increasing density is critically reviewed, it appears to be based on firm experimental evidence, confirmed by in vivo demonstration of decreasing survival of cells of increasing age. Analysis of studies using buoyant density gradients reveals that this technique can easily distinguish the single exponential slope of decline for those cell components that change progressively throughout the red cell life span from the biphasic decline of those that decrease drastically at the reticulocyte-mature red cell transition. The view that the aging of the red cell and its removal from the circulation result from a progressive series of events during the 120 days of its life span appears to be the most consistent with the available data. Density separation, validated by much experimental evidence, remains a most useful technique for the study of the mechanism of aging of the red cell. PMID- 8416297 TI - Study of glucose-6-phosphate dehydrogenase: history and molecular biology. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency was discovered in the 1950s. The history of the development of knowledge about G6PD deficiency is reviewed here. In the first decade after its discovery, the clinical manifestations of G6PD deficiency began to be understood. In the second decade, attention was focused on the degree of variability of this enzyme and the distinction of the various biochemical variants from one another. In the last decade, it has been possible to understand the mutations that effect this enzyme at the DNA level. Some 40 different mutations have now been characterized. Analysis of these mutations indicates that, while diversity sometimes exists within a mutation considered biochemical homogeneous, more often variants thought to be distinct prove to be identical. The study of G6PD mutations is beginning to provide insight into structure-function relationships. PMID- 8416298 TI - Bioactivity of heme and its containment. PMID- 8416299 TI - Cobalamin absorption and hematologic status after two types of gastric surgery for obesity. AB - A series of morbidly obese patients was treated surgically with a gastric bypass, and a subsequent series received a vertical banded gastroplasty. To compare some of the nutritional effects of these two procedures we measured serum vitamin B12 levels, absorption of food vitamin B12, frequency of microcytosis of erythrocytes, and frequency of anemia at one or more years after surgery. Patients with a gastric bypass showed greater weight loss, a greater frequency of microcytosis and anemia, more frequent subnormal serum levels of vitamin B12, and impressive failure to absorb food vitamin B12. Boiling the food containing vitamin B12 led to increased absorption. PMID- 8416300 TI - A concise review: iron absorption--the mucin-mobilferrin-integrin pathway. A competitive pathway for metal absorption. AB - Newly identified iron binding proteins isolated from rat duodenal homogenates permit better understanding of iron absorption. Mucins bind iron at acid pH to keep iron soluble and available for absorption at the more alkaline pH of the duodenum; this explains iron deficiency following prolonged achlorhydria. Integrin (90/150 kD) was identified on the absorptive surface of enterocytes in association with radioiron and is believed to facilitate transit of iron through the microvillous membrane. Mobilferrin, a 56 kD iron binding protein, was isolated from enterocyte cytosol. It coprecipitates with integrin and appears in close association with integrins in the apical cytoplasm. We postulate it accepts dietary iron from integrin and acts as the shuttle protein for iron in the cytoplasm. Since iron in enterocytes remains in equilibrium with body stores, we postulate mucosal iron uptake is regulated by the number of iron binding sites either occupied or unoccupied by iron on mobilferrin. Iron repletion of enterocytes from body stores is accomplished via transferrin receptors on the posterolateral membranes of enterocytes. Increased transfer of iron from blood into absorptive enterocytes occurs in iron replete animals to inhibit mucosal uptake of dietary iron. Little transfer of iron from plasma to enterocytes occurs in iron deficiency. Enhanced mucosal transfer of iron into the body occurs with increased body need for iron. The exact mechanism for mucosal transfer of iron into the plasma has not been defined but may also be mediated by an integrin. PMID- 8416301 TI - Concise review: methemoglobinemia. AB - The ferrous iron of hemoglobin is exposed continuously to high concentrations of oxygen and, thereby, is oxidized slowly to methemoglobin, a protein unable to carry oxygen. To restore hemoglobin function, methemoglobin (ferrihemoglobin) must be reduced to hemoglobin (ferrohemoglobin). Under physiological conditions, methemoglobin reduction is accomplished mainly by red cell NADH-cytochrome b5 reductase (NADH-methemoglobin reductase) so efficiently that there is insignificant amounts of methemoglobin in the circulating blood. However, should methemoglobin formation be increased--e.g., due to the presence of oxidant drugs, or an abnormal methemoglobin not amenable to reduction (hemoglobin M), or a deficiency in red cell cytochrome b5 reductase--methemoglobinemia will result. Most methemoglobinemias have no adverse clinical consequences and need not be treated. Under certain conditions, such as exposure to large amounts of oxidant or in young infants, rapid treatment is necessary. In hereditary cytochrome b5 deficiency, treatment is often directed at improving the poor cosmetic effect of persistent cyanosis with the minimum amount of drugs to give satisfactory clinical results. PMID- 8416302 TI - Hepatic iron stores and plasma ferritin concentration in patients with sickle cell anemia and thalassemia major. AB - To examine the relationship between hepatic iron stores and plasma ferritin concentration in individuals treated with red cell transfusion and iron chelation therapy, 37 patients with sickle cell anemia and 74 patients with thalassemia major were studied. In each patient, hepatic iron stores were measured by an independently validated noninvasive magnetic method, and plasma ferritin was determined by immunoassay. The correlation between hepatic iron and plasma ferritin was significant both in patients with sickle cell anemia (R = 0.75, P < 0.0001) and in those with thalassemia major (R = 0.76, P < 0.0001). Regression analysis showed no significant difference between the two groups in the linear relationships between hepatic iron stores and plasma ferritin. Considering all 111 transfused patients as a group, the coefficient of correlation between hepatic iron stores and plasma ferritin was highly significant (R = 0.76, P < 0.0001). Regression analysis found that variation in body iron stores, as assessed by magnetic determinations of hepatic iron, accounted for only approximately 57% of the variation in plasma ferritin, suggesting that the remainder was the result of other factors, such as hemolysis, ineffective erythropoiesis, ascorbate deficiency, inflammation, and liver disease. The 95% prediction intervals for hepatic iron concentration, given the plasma ferritin, were so broad as to make a single determination of plasma ferritin an unreliable predictor of body iron stores. Variability resulting from factors other than iron status limits the clinical usefulness of the plasma ferritin concentration as a predictor of body iron stores. PMID- 8416303 TI - Postnatal changes in the quantities of globin chains and hemoglobin types in two babies with Hb H disease. AB - We have studied two babies with Hb H disease from birth to about six months of age and analyzed the changes in the relative quantities of the five globin chains (zeta, alpha, beta, G gamma, A gamma) and the four hemoglobins (Hb F, Hb A, Hb Bart's, Hb H) using different high performance liquid chromatography procedures. The types of Hb H disease were -(SEA)/-alpha(3.7 kb) and -(Fil)/-alpha(3.7 kb); the larger -(Fil) deletion includes the functional zeta 2-globin gene, explaining the higher zeta chain level in the baby with the -(SEA)/-alpha(3.7 kb) type. The functional hemoglobin level at birth (Hb A+Hb F) was 11 to 12 g/dl with 3 to 4 g/dl Hb Bart's (gamma 4). Only 5% of the "fast-moving" hemoglobin was Hb H (beta 4). The level of Hb F at birth was low (less than 50% of the total Hb A+Hb F). After birth, the alpha and gamma chain production decreases rapidly resulting in a severe anemia (total functional hemoglobin approximately 7 g/dl) at 30 to 60 days postnatally, improving gradually to 8.5-9.5 g/dl at age of three months. The preferential formation of Hb A over Hb F at birth, and presumably prenatally, has the advantage that the level of the highly unstable Hb H is kept low; it also results in low levels of Hb F impairing the oxygen transfer capability of the fetal blood. PMID- 8416304 TI - Sickle cell anemia is a multigene disease: sickle painful crises, a case in point. PMID- 8416305 TI - Clinical and genetic studies of renal cell carcinomas in a family with a constitutional chromosome 3;8 translocation. Genetics of familial renal carcinoma. AB - OBJECTIVE: To describe the clinical course and genetic studies of renal carcinoma in members of a family with the constitutional chromosome translocation, t(3;8) (p14;q24). DESIGN: A follow-up study that updates our 1979 report of renal carcinoma in 10 of these relatives. SETTING: A cancer center and university hospital. PATIENTS: Members of the family, including five carriers of the 3;8 translocation who were in remission of renal cancer. MEASUREMENTS: Clinical follow-up of the family and genetic analyses of the renal cancer specimens of three patients. RESULTS: Renal carcinoma recurred in all five patients in the family at 1 to 16 years of follow-up. Three patients have died of renal cancer, and two are in a second remission. The renal cancers from three family members consistently reveal loss of the entire derivative chromosome 8, which bears the chromosome 3p segment spanning band p14 to the telomere. In contrast, no genetic change was detected in the derivative chromosome 3 or in normal chromosomes 3 and 8. CONCLUSIONS: This family illustrates the importance of clinical follow-up of patients with a hereditary cancer that can develop at multiple foci and recur over time. The inherited 3;8 translocation and loss of the translocated distal chromosome 3p in tumor specimens of family members may help localize the gene or genes involved in the pathogenesis of both familial and sporadic renal carcinoma. PMID- 8416306 TI - The use of bile acid sequestrants to lower serum thyroid hormones in iatrogenic hyperthyroidism. PMID- 8416307 TI - Reversible neurologic toxicity in patients treated with standard-dose fludarabine phosphate for mycosis fungoides and chronic lymphocytic leukemia. PMID- 8416308 TI - Transmission of infection by gastrointestinal endoscopy and bronchoscopy. AB - OBJECTIVE: To review reports on the transmission of infections by flexible gastrointestinal endoscopy and bronchoscopy in order to determine common infecting microorganisms, circumstances of transmission, and methods of risk reduction. DATA SOURCES: Relevant English-language articles were identified through prominent review articles and a MEDLINE search (1966 to July 1992); additional references were selected from the bibliographies of identified articles. STUDY SELECTION: All selected articles related to transmission of infection by gastrointestinal endoscopy or bronchoscopy; 265 articles were reviewed in detail. DATA SYNTHESIS: Two hundred and eighty-one infections were transmitted by gastrointestinal endoscopy, and 96 were transmitted by gastrointestinal endoscopy, spectrum of these infections ranged from asymptomatic colonization to death. Salmonella species and Pseudomonas aeruginosa were repeatedly identified as the causative agents of infections transmitted by gastrointestinal endoscopy, and Mycobacterium tuberculosis, atypical mycobacteria, and P. aeruginosa were the most common causes of infections transmitted by bronchoscopy. One case of hepatitis B virus transmission via gastrointestinal endoscopy was documented. Major reasons for transmission were improper cleaning and disinfection procedures; the contamination of endoscopes by automatic washers; and an inability to decontaminate endoscopes, despite the use of standard disinfection techniques, because of their complex channel and valve systems. CONCLUSIONS: The most common agents of infection transmitted by endoscopy are Salmonella, Pseudomonas, and Mycobacterium species. To prevent endoscopic transmission of infections, recommended disinfection guidelines must be followed, the effectiveness of automatic washers must be carefully monitored, and improvements in endoscope design are needed to facilitate effective cleaning and disinfection. PMID- 8416309 TI - Effect of antihypertensive therapy on the kidney in patients with diabetes: a meta-regression analysis. AB - OBJECTIVE: To assess the relative effect of different antihypertensive agents on proteinuria and renal function in patients with diabetes. DATA SOURCES: We used MEDLINE and bibliographies in recent articles to identify studies of the effects of antihypertensive agents on renal function in patients with diabetes. STUDY SELECTION: We selected 100 controlled and uncontrolled studies that provided data on renal function, proteinuria, or both, before and after treatment with an antihypertensive agent. DATA EXTRACTION: Data on blood pressure, renal function, proteinuria, patient characteristics (for example, age, sex, and type of diabetes), and study design (for example, random allocation and the use of a placebo) were extracted from selected studies. DATA SYNTHESIS: Multiple linear regression analysis indicated that angiotensin-converting enzyme (ACE) inhibitors decreased proteinuria independent of changes in blood pressure, treatment duration, and the type of diabetes or stage of nephropathy, as well as study design (P < 0.0001). Reductions in proteinuria from other antihypertensive agents could be entirely explained by changes in blood pressure. Blood pressure reduction in itself was associated with a relative increase in glomerular filtration rate (regression coefficient [+/- SE], 3.70 +/- .92 mL/min for each reduction of 10 mm Hg in mean arterial pressure; P = 0.0002); however, compared with other agents, ACE inhibitors had an additional favorable effect on glomerular filtration rate that was independent of blood pressure changes (3.41 +/- 1.71 mL/min; P = 0.05). CONCLUSION: Angiotensin-converting enzyme inhibitors can decrease proteinuria and preserve glomerular filtration rate in patients with diabetes. These effects occur independent of changes in systemic blood pressure. PMID- 8416310 TI - Correctional health care: a public health opportunity. AB - The approximately 1.2 million inmates in U.S. correctional institutions have a high prevalence of communicable diseases, such as human immunodeficiency virus (HIV) infection, tuberculosis, hepatitis B virus infection, and gonorrhea. Before their incarceration, most inmates had limited access to health care, which, together with poor compliance because of lifestyle, made them difficult to identify and treat in the general community. Because of the high yearly turnover (approximately 800% and 50% in jails and prisons, respectively), the criminal justice system can play an important public health role both during incarceration and in the immediate postrelease period. A public policy agenda for criminal justice should include an epidemiologic orientation, as well as resources for education, counseling, early detection, and treatment. Taking advantage of the period of confinement would serve both the individual and society by controlling communicable diseases in large urban communities. PMID- 8416311 TI - Assessment of cardiovascular risk: a return to basics. PMID- 8416312 TI - Outpatient liver biopsy: how safe is it? PMID- 8416313 TI - Estrogen and postmenopausal osteoporosis. PMID- 8416314 TI - Estrogen and postmenopausal osteoporosis. PMID- 8416315 TI - Estrogen and postmenopausal osteoporosis. PMID- 8416316 TI - Electronic thermometers and nosocomial infections. PMID- 8416317 TI - Thrombolysis and ventricular tachycardia. PMID- 8416318 TI - Sarcoidosis and sickle cell disease. PMID- 8416319 TI - Correctional health care. PMID- 8416320 TI - The bellman always rings thrice. PMID- 8416321 TI - Preventing anticoagulant-related bleeding. PMID- 8416322 TI - Value of the history and physical in identifying patients at increased risk for coronary artery disease. AB - OBJECTIVE: To determine whether information from the physician's initial evaluation of patients with suspected coronary artery disease predicts coronary anatomy at catheterization and 3-year survival. DESIGN: Prospective validation of regression model estimates in an outpatient cohort. SETTING: University medical center. PATIENTS: A total of 1030 consecutive outpatients referred for noninvasive testing for suspected coronary artery disease; 168 of these patients subsequently underwent catheterization within 90 days. MEASUREMENTS: Information from the initial history, physical examination, electrocardiogram, and chest radiograph was used to predict coronary anatomy (the likelihood of any significant coronary disease, severe disease [left main or three-vessel], and significant left main disease) among 168 catheterized patients and to estimate 3 year survival among all patients. These estimates were compared with those based on treadmill testing. Cardiovascular testing charges were calculated for all patients. RESULTS: Predicted coronary anatomy and survival closely corresponded to actual findings. Compared with the treadmill exercise test, initial evaluation was slightly better able to distinguish patients with or without any coronary disease and was similar in the ability to identify patients at increased risk for dying or with anatomically severe disease. Based on arbitrary definitions, 37% to 66% of patients were at low risk and responsible for 31% to 56% of the charges for cardiovascular testing. CONCLUSIONS: The physician's initial evaluation, despite the subjective nature of much of the information gathered, can be used to identify patients likely to benefit from further testing. The development of strategies for cost-conscious quality care must begin with the history, physical examination, and simple laboratory testing. PMID- 8416323 TI - Dietary risk factors for the incidence and recurrence of colorectal adenomatous polyps. A case-control study. AB - OBJECTIVE: To investigate the association of dietary factors with the incidence and recurrence of colorectal adenomatous polyps. DESIGN: Two case-control studies. SETTING: Three university-based colonoscopy practices in New York City. PATIENTS: For the incidence study, 286 patients with pathologically confirmed incident adenomatous polyps (162 men and 124 women) were compared with 480 controls (210 men and 270 women) with no current or previous neoplasia. For the recurrence study, 186 patients with recurrent polyps (130 men and 56 women) were compared with 330 controls (187 men and 143 women). These patients had a history of polyps but no current neoplasia. MEASUREMENTS: Structured interviews using the Block food frequency questionnaire were conducted on all participants and were compared over quartiles of crude nutrient intake of total and saturated fat; fiber; protein; carbohydrates; carotene; vitamins A, C, and E; and various food groups. Data were adjusted for age, Quetelet index, and caloric intake by multiple logistic regression for men and women separately. RESULTS: For incident polyps, elevated adjusted odds ratios (high to low quartile) for women were found for saturated fat (odds ratio, 2.3; 95% CI, 0.9 to 5.8) and the ratio of red meat to chicken and fish intake (odds ratio, 1.9; CI, 1.0 to 3.6). Protective associations were observed for fish and chicken (odds ratio, 0.6; CI, 0.3 to 1.2) and vitamin A intake (odds ratio, 0.4; CI, 0.2 to 0.9). Among women, recurrent polyps showed an association with total dietary fat (odds ratio, 4.4; CI, 1.0 to 19.5), saturated fat (odds ratio, 3.8; CI, 1.0 to 14.9; P = 0.15 for the trend), and total fiber (odds ratio, 0.2; CI, 0.1 to 0.5; P = 0.01) and a borderline association with carbohydrates (odds ratio, 0.3; CI, 0.1 to 1.3; P = 0.10). No consistent relations were observed for men, although increased caloric intake increased the risk for incidence and recurrence in both men and women. CONCLUSIONS: These findings are consistent with previously described relations between diet and the incidence of colon cancer and suggest that, at least in women, dietary modification may be warranted in patients diagnosed with polyps. PMID- 8416324 TI - Outcome of patients hospitalized for complications after outpatient liver biopsy. AB - OBJECTIVE: To determine the safety of outpatient liver biopsies by analyzing the outcome of patients hospitalized for complications. DESIGN: Retrospective review. SETTING: Large clinic referral center. PATIENTS: All patients admitted after outpatient liver biopsy at the Mayo Clinic from 1 April 1989 to 1 April 1991. RESULTS: During this period, 405 outpatients underwent biopsy. Of the 405 patients, 13 (3.2%) were admitted with complications after biopsy. Five patients (38%) were admitted with persistent localized pain, five (38%) with orthostatic hypotension, one (8%) with both pain and hypotension, one (8%) with peritoneal signs, and one (8%) with lightheadedness but no orthostatic changes. All complications were noted within 3 hours after the biopsy. Bleeding, potentially the most serious complication, was radiographically defined in 5 of the 13 patients (38%) admitted. Only two patients, however, required blood transfusions. No patient required invasive management such as surgery or chest tube placement. The average length of the hospital stay was 1.5 days. CONCLUSION: Complications after outpatient liver biopsy occur early and rarely require invasive management. Outpatient liver biopsy is safe when done on carefully selected patients in a setting that provides close observation for at least 3 hours after liver biopsy. PMID- 8416325 TI - Beta-blockers after myocardial infarction: influence of first-year clinical course on long-term effectiveness. AB - OBJECTIVE: To develop a strategy for evaluating drug efficacy over time that accounts for heterogeneous clinical courses evolving after initiation of therapy and to demonstrate its use in assessing the long-term therapeutic benefit of propranolol after myocardial infarction. DESIGN: Analysis of data from the Beta Blocker in Heart Attack Trial (BHAT), a randomized, double-blind, placebo controlled trial that enrolled patients from 1978 to 1980 and followed participants for vital status to April 1982. SETTING: Thirty-one clinical centers in the United States and Canada. PATIENTS: Eligible patients included 3297 men and women 30 to 69 years of age who survived 1 year after trial entry. INTERVENTION: Patients were classified as being on treatment at 12 months after randomization if they were receiving beta-blocker therapy at the 12-month visit and off treatment if they were not receiving beta-blocker therapy at that time. OUTCOME MEASURE: Vital status evaluated at 720 days of follow-up. RESULTS: A total of 2914 patients (88%) was classified as being at lower risk (strata I and II). For these patients, survival curves by treatment at 12 months were virtually indistinguishable. Among the 383 patients categorized as being at high risk on the basis of recurrent ischemic events, arrhythmias, congestive heart failure, or severe comorbidity during the first 12 months, the use of beta-blockers was associated with a 43% proportional decline in the subsequent risk for death (P = 0.01 by log-rank test). CONCLUSIONS: In patients who survived to 1 year with low- to moderate-risk clinical courses, beta-blocker therapy did not have long-term beneficial effect. In contrast, among patients who had a high-risk clinical course during the first year, beta-blockers significantly reduced mortality in the follow-up period. PMID- 8416326 TI - Discordant results of visual and quantitative estimates of stenosis severity before and after coronary angioplasty. AB - The ability to accurately estimate the severity of epicardial coronary stenoses is critical in the assessment of the immediate and long-term results of percutaneous transluminal coronary angioplasty (PTCA). We prospectively compared visual estimates, performed by experienced interventional cardiologists, with computerized quantitative angiographic measurements of stenosis severity in a group of patients (n = 305) before, immediately after and 6 months after PTCA. Before PTCA the visual estimate of the mean (+/- SD) percentage stenosis severity, 80.6 (+/- 9.7)%, was significantly (P < 0.001) higher than the equivalent value, 73.4 (+/- 11.1)%, obtained with use of quantitative angiography. Immediately after PTCA the visual estimate of the mean residual stenosis, 18.8 (+/- 12.3)%, was significantly (P < 0.0001) lower than the equivalent quantitative estimate, 37.4 (+/- 14)%. Additionally, the residual stenosis was more frequently (18% vs. 3%) classified as significant (> 50%) by quantitative angiography. At follow-up, quantitative measurements of stenosis severity showed a Gaussian distribution with a mean of 54.8 (+/- 21)%, whereas visual estimates had a bimodal distribution with populations greater than 70% and less than 50%. Visual estimates provide an inaccurate assessment of the immediate and medium term results of PTCA. Quantitative measurements suggest that the immediate results of PTCA are frequently misclassified as successful by the operator. Conversely, restenosis rates are underestimated by the operators suggesting that a more objective method must be used in trials to assess the impact of new therapies on the rate of restenosis. PMID- 8416327 TI - Endomyocardial biopsy in heterotopic heart transplant recipients via the femoral vein. AB - The results of endomyocardial biopsy (EMB) via the femoral vein in heterotopic heart transplant recipients were retrospectively analyzed and compared with those obtained using the right internal jugular vein approach. A total of 139 EMB were performed in 8 patients using the femoral (35) or the jugular (104) approach. Twenty three (64.7%) of the procedures performed via the femoral vein were part of the yearly hemodynamic and coronary artery study, and 12 (35.3%) constituted a routine postoperative evaluation of the myocardium rejection state in patients with imperviousness of the right internal jugular vein. Comparing the results obtained with the femoral approach, we observed a higher overall success rate (94.3 vs 88.5%, NS) and obtained more samples that were useful for histologic evaluation (95.5 vs 85.9%, NS); with the jugular procedure, the fragments were significatively larger in diameter (1.28 +/- 0.55 vs 1.61 +/- 0.85 mm, mean +/- SD) and in area (1.49 +/- 1.16 vs 2.28 +/- 2.24 mm2, mean +/- SD). No cardiac or local complications were noted when the femoral approach was used, while two attempts to perform biopsy via the jugular vein resulted in obstruction of this vessel. Our data suggest that the femoral venous approach for endomyocardial biopsy in heterotopic heart transplant recipients is a valid alternative to the more commonly used routes. PMID- 8416328 TI - Pulmonary wedge angiography for prediction of pulmonary vascular disease in Down syndrome. AB - We performed high resolution pulmonary wedge angiography (PWA) and conventional hemodynamics to predict the reversibility of structural pulmonary vascular disease. Sixty-one pulmonary wedge angiograms were performed on 41 patients with intracardiac shunts and Down syndrome (median age 8 months). Balloon occlusion wedge angiograms were analyzed for (1) monopedial branches from the distal 10 mm of muscular arteries, (2) capillary blush, (3) tapering indices, and (4) tortuosity. Twenty-five patients had open lung biopsy, graded by the Health Edwards classification, and analyzed morphometrically. Pulmonary vascular resistance of > or = 6 units was 100% sensitive and 94% specific for Heath Edwards Grade III-IV. A monopedial count < 3 vessels was 83% sensitive and 100% specific for Heath Edwards Grade III-IV. Abnormal capillary blush was 83% sensitive and 69% specific for Heath Edwards Grade III-IV. Tapering indices and tortuosity showed no significant correlation with lung biopsy. A combination of pulmonary vascular resistance < 6 units, monopedial count > or = 3, and normal capillary blush was 100% sensitive and specific for Heath Edwards Grade 0-II, and a combination of pulmonary vascular resistance > or = 6 units, monopedial count < 3, and abnormal capillary blush was 100% sensitive and specific for Heath Edwards Grade III-IV. Using the 3 criteria, Heath Edwards Grade was accurately predicted in 17 patients. In 4 patients, only 2 criteria were available. Morphometric analysis showed an inverse relationship between the lowest monopedial count and the number of occlusive vessels per cm of tissue, r = -0.74 p < 0.001. Arteries showing intimal and/or medial thickening causing > 90% luminal narrowing were scored as "occlusive." These results show that when the hemodynamic and pulmonary wedge angiography data are concordant, the structural changes of pulmonary vascular disease can be accurately predicted and lung biopsy may be avoided. PMID- 8416329 TI - Early ambulation following diagnostic 7-French cardiac catheterization: a prospective randomized trial. AB - There is a paucity of randomized studies concerning transfemoral cardiac catheterization and its complications, in particular that of 7F catheterization. Accordingly, we conducted a prospective, randomized trial comparing early ambulation (group A) 6 hr after diagnostic 7F cardiac catheterization versus late ambulation (group B) the following morning. A total of 273 patients were randomized in the study; 142 in group A and 131 in group B (NS). Except for a difference in the indications for catheterization, the baseline and procedure related parameters were similar between the 2 groups. Early hematoma (formed within 6 hr) developed in 6 (4%) and 7 (5%) patients in groups A and B, respectively (NS). Similarly, there was no difference in the incidence of late hematoma formation (2% in each group). All hematomas detected were small and required no surgical intervention or extension of hospital stay. Our data showed that early ambulation following 7F left heart catheterization is feasible and safe. The access site complication rate is acceptably low and minor in nature. PMID- 8416330 TI - Acute myocardial infarction complicating urokinase infusion for total saphenous vein graft occlusion. AB - Saphenous vein graft occlusions have been successfully treated with extended urokinase infusions. We report a case of myocardial infarction complicating this treatment. A review of reported cases suggests that this complication may not be uncommon. The optional drug, dose, and infusion technique for intra-graft lytic therapy has not been determined. The costs, risks, and difficulty of this technique may limit its application. PMID- 8416331 TI - Use of excimer laser in the treatment of chronic total occlusion of a coronary artery that cannot be crossed with a balloon catheter. AB - The inability to advance a balloon across a chronically occluded coronary artery that has been crossed with a guidewire occurs in 5% of patients with chronic occlusions. In this report, a new indication for the excimer laser is described: the recanalization of a chronic occlusion that has been crossed with a guidewire that cannot be crossed with a balloon catheter. PMID- 8416332 TI - Coronary artery rupture and pseudoaneurysm formation resulting from percutaneous coronary angioscopy. AB - We describe a case in which coronary angioscopy was complicated by inability to deflate the device's occlusion balloon. Rapid over-inflation to rupture the balloon resulted in massive dissection of the artery, pseudoaneurysm formation, and ultimately coronary bypass. While the cause of failure of balloon deflation remains obscure, deliberate over-inflation to cause rupture may be hazardous. PMID- 8416333 TI - Interpretation of cardiac pathophysiology from pressure waveform analysis: simultaneous left and right ventricular pressure measurements. AB - In addition to demonstrating constrictive and restrictive cardiac physiology, simultaneous right and left ventricular pressure measurements can be helpful to identify various aspects of myocardial dysfunction. Intracardiac conduction defects will displace the right ventricular pressure under the left ventricular pressure upstroke and identify differences in the timing of ventricular contraction. Right ventricular dysfunction will also produce abnormal right ventricular pressure waveforms which may overlap left ventricular pressure and contribute to abnormalities in right atrial and ventricular pressure waveforms. PMID- 8416334 TI - Limitations of the zero crossing detector in the analysis of intracoronary Doppler: a comparison with fast Fourier transform analysis of basal, hyperemic, and transstenotic blood flow velocity measurements in patients with coronary artery disease. AB - The current clinical standard for the analysis of intracoronary Doppler signals is the application of a zero-crossing (ZC) detector. However, the accuracy of the method is questionable, especially in areas of disturbed flow, as confirmed by in vitro studies, animal experiments, and intraoperative observations. The aim of this study is the comparison of a conventional ZC detector and a custom-designed spectral analyzer (fast Fourier transform, FFT) in the analysis of intracoronary Doppler signals obtained in 19 patients undergoing coronary angioplasty. A 3F catheter with an end-mounted Doppler ceramic crystal was placed over an 0.014" guidewire in a normal or near-normal segment proximal to the lesion to be dilated. The Doppler signal was recorded before and after intracoronary infusion of 12.5 mg of papaverine. In 9 patients high flow velocities could be recorded when the catheter was advanced across the stenosis. The blood flow velocity measurements obtained with ZC were significantly lower than the maximal FFT flow velocity measurements (16 +/- 12 cm/s vs. 29 +/- 18 cm/s, p < .001). In all the conditions of Doppler signal acquisition (baseline, hyperemia, stenosis) a large scattering of the signed differences between corresponding measurements was observed. The standard deviation of the difference ZC-FFT was +/- 11 cm/s and +/- 5 cm/s for the maximal and mean FFT flow velocity, corresponding in both cases to +/- 37% of the mean of the ZC and FFT measurements.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416335 TI - Platelet adhesion/aggregation in an in vitro model of coronary artery stenosis. AB - Platelet adhesion/aggregation (PAA) at a site of coronary artery stenosis is believed to be a process strongly modulated by local shear rates and the functional state of neighboring endothelium. One purpose of the present work, therefore, is to describe an in vitro model for the direct imaging of such PAA. Another is to apply the model to the question as to whether the use of nonionic vs. ionic contrast media (CM) in the presence of vascular endothelium contributes to PAA at the stenosis site. Toward these ends, we utilized a special flow chamber which incorporates a monolayer of endothelial cells (ECs), a step 66% flowpath constriction at a site preadsorbed with microfibrillar collagen, and arterial shear rates. By epifluorescence microscopy and digital image analysis of video recordings, PAA was found to be greater with dysfunctional ECs (pretreated with lysine acetylsalicyclate) than with normal ECs, thereby confirming a modulatory role in PAA of functionally intact ECs. When nonionic (iohexol) or ionic (ioxaglate, diatrizoate) CM was added to the flowing blood at a concentration of 20% by non-red cell volume, PAA was inhibited in the order diatrizoate > ioxaglate > iohexol > saline control. No inhibition by any CM was seen, however, when chamber prefill culture medium containing 20% by volume CM was displaced by CM-free blood, in simulation of bolus administration of CM. In terms of inhibition of PAA during percutaneous transluminal coronary angioplasty (PTCA), therefore, our model provides a conceptual basis by which one may anticipate in flowing blood no clear benefit of ionic over nonionic CM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416336 TI - Laser facilitated angioplasty and thrombolysis in acute myocardial infarction complicated by prolonged or recurrent chest pain. AB - To date, application of laser angioplasty in acute myocardial infarction (MI) has not been reported. In nine patients with acute myocardial infarction complicated by continuous or recurrent severe ischemia and chest pain, a mid-infrared, solid state, pulse-wave holmium/thulium:YAG coronary laser was applied. In six of these patients the laser was specifically utilized for the purpose of coronary thrombolysis. In each case a guidewire was placed across the stenosis and a multifiber laser catheter was utilized, emitting 250-600 mJ/pulse at 5 Hz, followed by adjunctive balloon angioplasty. Laser success (defined as ability to cross the lesion, reduction of > or = 20% in stenosis and thrombolysis when a thrombus is present) was achieved in all patients. Final angiograms revealed residual stenosis < or = 30%, adequate thrombolysis and no major complication (MI, perforation, emergency CABGS, CVA, death) in each patient. Clinically, all nine patients improved, survived the acute infarction and were discharged. This initial clinical experience demonstrates the feasibility and safety of holmium/thulium:YAG laser application in thrombolysis and plaque ablation in selected patients who experience acute myocardial infarction complicated by prolonged or recurrent ischemia and chest pain. PMID- 8416337 TI - Parallel angioplasty dilatation catheter and guide wire: a new technique for the dilatation of calcified coronary arteries. AB - Percutaneous transluminal coronary angioplasty (PTCA) of heavily calcified rigid coronary arteries has decreased success and increased complication rates. Three cases are presented describing a new technique for the dilatation of severely calcified coronary arteries that were not dilatable by conventional angioplasty methods. This technique involves the use of a balloon dilatation catheter system parallel to a guide wire. PMID- 8416338 TI - Triple balloon-on-a-wire or "menage a trois" coronary angioplasty. AB - We have described here the triple balloon or "menage a trois" technique of coronary angioplasty of trifurcation lesions or closely approximated bifurcation lesions. Although the need for this technique is uncommon on anatomic grounds, it can facilitate excellent angiographic results. PMID- 8416339 TI - Direct deployment of the iliofemoral balloon expandable (Palmaz) stent utilizing a small (7.5 French) arterial puncture. AB - A new, smaller (7.5F) stent delivery sheath system is described for deployment of the iliofemoral large balloon expandable (Palmaz) stainless steel stent which allows a brachial or ipsilateral retrograde femoral or popliteal approach. The procedure described does not require predilatation of the lesion prior to stent deployment. PMID- 8416340 TI - Successful limb salvage after recanalization of an occluded infrapopliteal artery utilizing a balloon expandable (Palmaz-Schatz) stent. AB - This case describes the first reported successful management of an infrapopliteal artery intimal dissection utilizing the Palmaz-Schatz balloon expandable stent. Stent deployment relieved the compromised blood flow caused by the intimal flap which could not be corrected by prolonged balloon inflation or removed with directional atherectomy, and averted any emergency surgical procedure. PMID- 8416342 TI - Using a Touhy Borst system for flushing thin catheter lumens which do not have a hub for a syringe. PMID- 8416341 TI - Continuous cardiac output monitoring by the Fick method. AB - Current methods for longitudinal assessment of cardiac output in severely ill patients are intermittent only and in many respects appear unsatisfactory. We have developed a computerized on-line system for continuous Fick cardiac output monitoring, utilizing fiberoptic arterial and pulmonary arterial probes with a metabolic analyzer for VO2. In 15 patients, cardiac output ranged 1.9-6.8 L/min and 12 were within 5% of thermodilution values. Continuous output monitoring during interventions in two patients (saline infusion and coronary angioplasty) illustrate the utility of the technique. Two additional patients had unsatisfactory VO2 data due to low airflow velocity. The results of this pilot study suggest that cardiac output monitoring by the Fick method may have clinical and investigational uses in intensive care units and during cardiac catheterization or surgical procedures. PMID- 8416343 TI - Translumbar approach to coronary angiography. PMID- 8416344 TI - Latex allergy in children with myelodysplasia: a survey of Shriners hospitals. AB - We surveyed the incidence of recognized latex sensitivity in myelodysplasia patients in 16 Shriners Hospitals. The percentage of involved patients ranged from 0 to 22% (mean 5%). Twenty-two children had a significant anaphylactic reaction. We provide recommendations for preoperative prophylaxis and sources of listings of latex and latex-free hospital products and home products. PMID- 8416345 TI - Refracture of adolescent femoral shaft fractures: a complication of external fixation. A report of two cases. AB - Malunion is often a problem in adolescent femoral shaft fractures treated with traction and subsequent casting. We used external fixation as an alternative treatment method to control alignment and allow early mobilization. After fracture healing and fixator removal, two patients experienced refracture. This unusual complication is the basis of our report. Prolonged rigid or static fixation appears to be detrimental to sufficient callus formation and healing. PMID- 8416346 TI - Locke, Rousseau, and the modern surgeon. PMID- 8416347 TI - Relationship between acquired spastic talipes equinovarus and dystonia musculorum deformans. AB - Acquired spastic talipes equinovarus associated with dystonia musculorum deformans (DMD) presents orthopaedic surgeons with a difficult treatment problem. We reviewed the natural history, diagnostic evaluation, and treatment of this deformity in 12 patients. Two patients initially had the generalized form of dystonia, and 10 patients had the focal form (in which acquired spastic talipes equinovarus was the only manifestation). With time, five of these 10 developed the generalized form. A positive family history and age < 14 years at onset were prognostic of this progression. PMID- 8416348 TI - Split tibialis posterior tendon transfer and tendo-Achillis lengthening for spastic equinovarus feet. AB - Twenty-one patients with a minimum follow-up of 2 years who underwent combined split tibialis posterior tendon transfer and tendo-Achilles lengthening for equinovarus foot deformities were evaluated. The results in 15 of 18 ambulatory patients were graded as excellent or good; patients had marked improvement of their equinovarus foot deformity in both stance and swing phases of gait and became brace-free postoperatively. All non-ambulatory patients had a plantigrade foot. The complication rate was low, and patient satisfaction with the procedure was very high. In three patients, however, the procedure failed because of technical errors. We address the causes of failure and methods to avoid these errors. Preoperative computerized gait analysis does not appear to be essential in achieving a good result. PMID- 8416349 TI - Preoperative and postoperative assessment of surgical intervention for equinus gait in children with cerebral palsy. AB - We compared the effects of different methods of surgical correction of equinus gait in children with spastic cerebral palsy (CP) using pre- and postoperative measurements of gait, electromyography (EMG), range of motion (ROM), and dynamic ankle motion. Operative results significantly improved all variables that produced more normal ankle motion in subjects, although no differences were observed between surgical methods. We conclude that preoperative gait analysis can assist in surgical planning regardless of surgical method for Achilles lengthening and can provide an objective measure of results after surgical correction of equinus deformity. PMID- 8416350 TI - Long-term follow-up of hip subluxation in cerebral palsy patients. AB - Spastic subluxation of 64 hips in 45 patients with cerebral palsy (CP) was reviewed in a 19-year follow-up. Varus osteotomy prevented dislocation in all patients. All nine hips that later dislocated were in quadriplegic patients who had not received treatment or who had had muscle releases. Dislocated hips had the most degenerative arthritis, pain, and the least movement. Subluxated hips also had more degenerative arthritis but approximately the same level of pain as reduced hips. PMID- 8416351 TI - Incidence of spondylolisthesis in ambulatory cerebral palsy patients. AB - We prospectively studied 50 consecutive ambulatory cerebral palsy (CP) patients to determine the incidence of isthmic spondylolisthesis. In addition, we examined the relationship of hip flexion contractures to development of spondylolisthesis and low back pain. Three patients who had undergone previous spine operation were eliminated from the study group. Of the remaining 47 patients, one patient (2%) demonstrated an asymptomatic grade I spondylolisthesis. Another patient (2%) demonstrated spondylolysis without spondylolisthesis. Only six patients reported occasional low back pain. Pain did not correlate with increasing age, increasing hip flexion contracture, or decreasing sacrofemoral angle. The incidence of spondylolisthesis in this group of ambulatory CP patients with hip flexion contractures is similar to that in the general population. Hip flexion contractures did not predispose the group to spondylolisthesis or low back pain. Periodic screening of asymptomatic ambulatory CP patients for spondylolisthesis is not recommended. PMID- 8416352 TI - Evolution of femoral head deformity during the healing phase of Legg-Calve Perthes disease. AB - Progressive changes in the roundness of the femoral head were noted during the healing phase of hips with Legg-Calve-Perthes disease. Forty-nine of 136 hips studied became progressively rounder, and 15 hips became progressively flatter after onset of reossification of the femoral head. The femoral head was more likely to undergo progressive flattening in older patients, in those with more severe lateral pillar involvement, and in those with prolonged reossification. These changes occurred during a 3- to 4-year reossification period after cessation of treatment. PMID- 8416353 TI - Epiphyseal distraction for bony bridges: a biomechanical and morphologic study. AB - Partial physeal bridges were produced by resecting part of the lower femoral physis in 8-week-old rabbits. After 4 weeks, an external fixator, strain-gauged to measure distraction forces, was applied across the physis. Distraction was performed for 15 days: 1.0 mm/day medially and 0.5 mm/day laterally. Weekly radiographs were obtained. Animals were killed at 0, 3, and 6 weeks after distraction. Characteristic force patterns were evident if the bridge fractured. Medial fixator forces just before bridge fracture were very high (mean 89 N, SD 28 N), although forces in the lateral fixator were low (mean 25 N, SD 15 N). All animals killed 3 and 6 weeks after distraction ceased showed complete physeal closure. Results suggest that distraction epiphysiolysis may have high potential for producing premature physeal fusion. PMID- 8416354 TI - Metatarsal epiphyseal bracket: treatment by central physiolysis. AB - Longitudinal epiphyseal bracket (LEB) is a rare ossification anomaly in which an epiphysis brackets the diaphysis of a phalanx, metacarpal, or metatarsal. This abnormal epiphysis tethers longitudinal growth, resulting in a shortened and oval shaped bone. Four patients with five LEBs were treated by central physiolysis and followed for a mean of 6 years. The patients had significant hallux varus deformity. Three patients had duplicated great toes, and two had tibia hemimelia significant enough to require epiphysiodesis as they neared adolescence. Resection of the LEB allowed the proximal and distal epiphysis to resume untethered growth. Silastic or methyl methacrylate was placed over the resected physis to prevent bony rebridging. The associated hallux varus deformity was corrected by capsulorrhaphy and K-wire fixation. In all patients, the metatarsal resumed longitudinal growth and correction of the hallux varus was maintained. PMID- 8416355 TI - Closed femoral shortening. AB - Eighteen patients underwent closed intramedullary femoral shortening at a children's teaching hospital. Their average age was 16 years 2 months (range 14 18 years). Clinical and radiographic union was obtained in < or = 3 months in all patients. Shortening averaged 4.4 cm (range 3-5 cm). Complications included one case of acute respiratory distress and three cases of fixation loss. We describe the procedure for closed intramedullary rodding without use of a fracture table and recommend (a) that reaming proceed slowly through an enlarged proximal portal, and (b) that in all cases nails be interlocked both proximally and distally. PMID- 8416356 TI - Femoral lengthening with the Barnes device. AB - Twenty femoral lengthenings were performed with a staged lengthening technique with the Barnes device. After the plate is attached to the femur, the femur is lengthened 2 cm initially and at two or more subsequent stages. Average length gained was 5.4 cm (range 2.5-8.3 cm), and there were no malunions or nonunions. There were two transient peroneal palsies, but no permanent nerve injuries. There were no pin tract problems, no chronic infections, and no knee subluxations. The complication rate with this technique compares favorably with those of other methods. PMID- 8416357 TI - Reconstruction of the proximal end of the femur after hematogenous osteomyelitis. AB - We report a method of surgical treatment of sequela of septic hips and osteomyelitis of the proximal end of the femur in seven children. Roentgenologic and clinical studies before operation showed unstable hip with loss of the femoral neck and head. Reconstruction osteotomy of remnants of the neck and/or head of the femur and a callus distraction procedure in the direction of the acetabulum with the Kalnberz external fixation device was performed. This allowed stabilization of the hip joint, new bone formation in anatomic place of the femoral head, and retention of hip joint function. PMID- 8416358 TI - Myoelectric and body-powered prostheses. AB - One hundred twenty below-elbow child amputees compared cosmetically identical myoelectric (MYO) and body-powered (BP) hands after wearing each for 3 months; 78% chose the MYO and 22% chose the BP. However, at 2-year follow-up, only 44% wore the MYO, 33% used a BP hand or hook, and 23% became nonwearers (NW). Eighty percent of children aged > or = 8 years used active prosthetic prehension for both the MYO and BP prostheses. Comparatively, only 30% aged < 8 years and 0% aged < 5 years demonstrated any active prosthetic prehension. Active BP wearers were clearly most pleased with the features of their prostheses. PMID- 8416359 TI - Skeletal age determination in young children: analysis of three regions of the hand/wrist film. AB - The determination of skeletal age in young children is useful in predicting eventual leg length discrepancies, but underreading of skeletal age may result in predicting the ultimate leg length to be much longer than it actually will be, with a correspondingly larger discrepancy as well. We evaluated 100 hand/wrist radiographs of 45 children aged between 14 months and 9.8 years. Separate readings were taken of the distal radius and ulna, the carpal bones, and the metacarpals and phalanges. The discrepancy between bone ages and chronologic age was significantly different between regions of the hand and between sexes. When determining bone age in children under age 10 years, care should be taken to consider the entire hand, perhaps with less emphasis on the carpal bones. PMID- 8416360 TI - Adolescent bunion deformity treated with double osteotomy and longitudinal pin fixation of the first ray. AB - Surgical correction of bunion deformity in adolescents is performed infrequently. Double osteotomy and longitudinal pin fixation of the first ray was performed on 15 feet. This procedure is technically easy, provides excellent correction and stability, and has a low rate of recurrence of deformity. Five numerical determinations were obtained from preoperative, intraoperative, and final follow up radiographs. They included intermetatarsal angle, first-fifth metatarsal angle, and metatarsophalangeal angle, first-second metatarsal distance, and length of the first and second metatarsals. The mean length of follow-up was 32.5 months. The operation is indicated for severe deformities and should be applicable to adults. PMID- 8416361 TI - Percutaneous reduction of displaced radial neck fractures in children. AB - Various techniques have been used for reduction of displaced radial neck fractures in children. Eighteen children with displaced radial neck fractures were treated by percutaneous reduction with a Steinmann pin after failing closed reduction. Reduction was successfully accomplished for 15 patients. The three patients failing percutaneous pin reduction had comminuted radial head and neck fractures (two patients) or a completely rotated and displaced radial head (one patient). Percutaneous pin reduction of angulated and displaced radial neck fractures is a simple, safe alternative to open reduction. PMID- 8416362 TI - Peritalar dislocations in children. AB - Five patients with peritalar dislocations were identified in a search for injuries of the talus. We report the mechanism, management, and outcome of their injuries and compare and contrast the injuries with the same injury in adults. To our knowledge, our series is the only small series of this rare injury in children. Four of our 5 patients had acceptable results with respect to pain and gait. In two patients, the peritalar dislocation was missed, necessitating a delay in management because more attention is focused on obvious fractures, when they exist. PMID- 8416363 TI - Vascular abnormalities of the extremities: clinical findings and management. AB - A retrospective review identified 41 patients with angiodysplastic lesions of the extremities. Twenty-two patients had a mass, 11 had limb length discrepancy and/or hemihypertrophy, and two had recurrent joint effusions. Enhanced computed tomography (CT) and magnetic resonance imaging (MRI) were most valuable for both diagnosis and preoperative planning. Many patients were diagnosed clinically and treated symptomatically. Twenty-nine patients (71%) required operation. Descriptive pathology and histologic diagnosis were not determinants of clinical outcome, whereas anatomic location and overall size were predictive of symptomatology. Subcutaneous hemangiomas irritated sensory nerves, intramuscular lesions mimicked compartment syndromes, intraarticular lesions caused recurrent hemarthroses, and periarticular or large lesions resulted in hypertrophy or limb length discrepancy. Simple excision under tourniquet control with incomplete exsanguination is possible but often incomplete (48% recurrence) owing to the invasive nature of the lesion. PMID- 8416364 TI - Determination of medial epicondylar epiphyseal angle for supracondylar humeral fractures in children. AB - A new radiographic angle, the medial epicondylar epiphyseal (MEE) angle, was studied in 100 anteroposterior (AP) radiographs of normal children aged 3-12 years. The mean value was 38.2 degrees +/- 4.17 degrees (SD) (range 25 degrees-46 degrees), and the MEE angle could be drawn satisfactorily in 96 radiographs. The MEE angle did not vary significantly from this value in 25 Jones' view radiographs (paired t test, t = 0.06). The MEE angle was useful in assessing the accuracy of reduction of 20 supracondylar fractures of the humerus with satisfactory final results. PMID- 8416365 TI - External fixation maintained until fracture consolidation in the skeletally immature. AB - In a prospective study, we investigated the effects of external fixation on fracture union in skeletally immature patients. Twenty-five fractures of the femur and tibia in 21 patients were fixed externally with the intention of leaving the fixator in place until the fractures consolidated or nonunion was established. The main indications were head injury and multiple trauma. One hundred percent of the fractures consolidated with the fixator in place; 45 of 50 joints had 100% motion, three had 95% motion, and two had 60% motion owing to scarring from traumatic wounds. Eight-four percent of the fractures lost no position in the fixator, and 16% lost < or = 5 degrees. PMID- 8416366 TI - The minor capsid protein VP1 of the autonomous parvovirus minute virus of mice is dispensable for encapsidation of progeny single-stranded DNA but is required for infectivity. AB - The two capsid proteins of minute virus of mice, VP1 and VP2, are generated from a single large open reading frame by alternate splicing of the capsid gene mRNA. Examination of the replication of a series of mutants that express only VP1, only VP2, or neither capsid protein demonstrates that VP2 is necessary for the accumulation and encapsidation of virus progeny single-stranded DNA. VP1 is dispensable for these functions but is required to produce an infectious virion. Virus that lacks VP1 binds to cells as efficiently as wild-type minute virus of mice but fails to initiate a productive infection. Because neither capsid protein is required for viral-DNA replication, these results suggest that virus lacking VP1 is blocked at a step during virus entry, subsequent to cell binding and prior to the initiation of DNA replication. PMID- 8416367 TI - Differentiated liver cell specificity of the second enhancer of hepatitis B virus. AB - Hepatitis B virus is a hepatotropic virus. Its replication and gene expression are mainly restricted to hepatocytes in the infective process. The viral gene expression thus provides a unique system with which to study the control of tissue-specific gene expression. We have previously reported the identification and characterization of the second enhancer (enhancer II) of hepatitis B virus. In this report, we further demonstrate that the minimal functional constituents of the second enhancer, box alpha and box beta, display liver cell and differentiation state specificity. Moreover, box alpha exhibits the same liver cell and differentiation state specificity when functioning as an upstream regulator for the basal core promoter. Gel shift experiments reveal a unique box alpha-binding protein, protein a, which is present only in differentiated liver cells, where enhancer II is functional. The converse is true for another box alpha-binding protein, protein f, which is present only in poorly differentiated liver cells and nonliver cells. The simplest hypothesis that explains these results is that protein a activates and/or protein f suppresses the enhancer and upstream regulator functions. Although C/EBP is a candidate for a transcription factor that interacts with box alpha or box beta, none of the binding factors identified in the gel shift assays, including protein a and protein f, is likely to be C/EBP because they differ from C/EBP in heat lability and sequence preference. PMID- 8416368 TI - Characterization of ribosomal frameshifting for expression of pol gene products of human T-cell leukemia virus type I. AB - For study of the pol gene expression of human T-cell leukemia virus type I (HTLV I), RNA was transcribed in vitro from proviral DNA and translated in rabbit reticulocyte lysates. This cell-free translation resulted in two major translation products representing the Gag and Gag-Pro polyproteins. By contrast, the Gag-Pro-Pol polyprotein could be readily observed only when translation was performed with mutant mRNA in which the protease (pro) reading frame was aligned to gag to eliminate the frameshifting event in the gag-pro overlap. The results indicated that two independent ribosomal frameshifting events are required for expression of the HTLV-I pol gene product. Studies with mutant DNAs facilitated the characterization of the primary structure of the HTLV-I mRNA responsible for the ribosomal frameshift in the pro-pol overlap and demonstrated that the frameshift occurs at the signal sequence UUUAAAC. Direct amino acid sequencing of the transframe protein localized the site of the frameshift to the asparagine codon AAC. PMID- 8416369 TI - DNA strand exchange catalyzed by proteins from vaccinia virus-infected cells. AB - Vaccinia virus infection induces expression of a protein which can catalyze joint molecule formation between a single-stranded circular DNA and a homologous linear duplex. The kinetics of appearance of the enzyme parallels that of vaccinia virus DNA polymerase and suggests it is an early viral gene product. Extracts were prepared from vaccinia virus-infected HeLa cells, and the strand exchange assay was used to follow purification of this activity through five chromatographic steps. The most highly purified fraction contained three major polypeptides of 110 +/- 10, 52 +/- 5, and 32 +/- 3 kDa. The purified protein requires Mg2+ for activity, and this requirement cannot be satisfied by Mn2+ or Ca2+. One end of the linear duplex substrate must share homology with the single-stranded circle, although this homology requirement is not very high, as 10% base substitutions had no effect on the overall efficiency of pairing. As with many other eukaryotic strand exchange proteins, there was no requirement for ATP, and ATP analogs were not inhibitors. Electron microscopy was used to show that the joint molecules formed in these reactions were composed of a partially duplex circle of DNA bearing a displaced single-strand and a duplex linear tail. The recovery of these structures shows that the enzyme catalyzes true strand exchange. There is also a unique polarity to the strand exchange reaction. The enzyme pairs the 3' end of the duplex minus strand with the plus-stranded homolog, thus extending hybrid DNA in a 3'-to-5' direction with respect to the minus strand. Which viral gene (if any) encodes the enzyme is not yet known, but analysis of temperature-sensitive mutants shows that activity does not require the D5R gene product. Curiously, v SEP appears to copurify with vaccinia virus DNA polymerase, although the activities can be partially resolved on phosphocellulose columns. PMID- 8416370 TI - Mutations in the cytoplasmic domain of human immunodeficiency virus type 1 transmembrane protein impair the incorporation of Env proteins into mature virions. AB - In-frame stop codons were introduced into the coding region of human immunodeficiency virus type 1 (HIV-1) transmembrane protein (gp41). Truncation of 147 amino acids from the carboxyl terminus of gp41 (TM709) significantly decreased the stability and cell surface expression of the viral Env proteins, while truncation of 104 amino acids (TM752) did not. Truncation of 43 or more amino acids from the carboxyl terminus of gp41 generated mutant viruses which were noninfectious in several human CD4+ T lymphoid cell lines and fresh peripheral blood mononuclear cells. Analysis of the noninfectious mutant virions revealed significantly reduced incorporation of the Env proteins compared with the wild-type virions. Comparable amounts of Env proteins were detected on the surfaces of wild-type- and TM752-transfected cells, suggesting that the structures of gp41 required for efficient incorporation of Env proteins were disrupted in mutant TM752. Truncation of the last 12 amino acids (TM844) from the carboxyl terminus of gp41 did not significantly affect the assembly and release of virions or the incorporation of Env proteins into mature virions. However, the TM844 virus had dramatically decreased infectivity compared with the wild-type virus. This suggests that the cytoplasmic domain of gp41 also plays a role in other steps of virus replication. PMID- 8416371 TI - A micromolar pool of antigenically distinct precursors is required to initiate cooperative assembly of hepatitis B virus capsids in Xenopus oocytes. AB - Assembly of hepatitis B virus capsid-like (core) particles occurs efficiently in a variety of heterologous systems via aggregation of approximately 180 molecules of a single 21.5-kDa core protein (p21.5), resulting in an icosahedral capsid structure with T = 3 symmetry. Recent studies on the assembly of hepatitis B virus core particles in Xenopus oocytes suggested that dimers of p21.5 represent the major building block from which capsids are generated. Here we determined the concentration dependence of this assembly process. By injecting serially diluted synthetic p21.5 mRNA into Xenopus oocytes, we expressed different levels of intracellular p21.5 and monitored the production of p21.5 dimers and capsids by radiolabeling and immunoprecipitation, by radioimmunoassay, or by quantitative enzyme-linked immunosorbent assay analysis. The data revealed that (i) p21.5 dimers and capsids are antigenically distinct, (ii) capsid assembly is a highly cooperative and concentration-dependent process, and (iii) p21.5 must accumulate to a signature concentration of approximately 0.7 to 0.8 microM before capsid assembly initiates. This assembly process is strikingly similar to the assembly of RNA bacteriophage R17 as defined by in vitro studies. PMID- 8416372 TI - Analysis of the adenovirus type 5 terminal protein precursor and DNA polymerase by linker insertion mutagenesis. AB - A series of adenovirus type 5 precursor terminal protein (pTP) and DNA polymerase (Ad pol) genes with linker insertion mutations were separately introduced into the vaccinia virus genome under the control of a late vaccinia virus promoter. The recombinant viruses were used for overexpression of the mutant genes in HeLa cells. In total, 22 different mutant pTP and 10 different Ad pol vaccinia virus recombinants were constructed, including some that expressed carboxyl-terminus truncated forms of both proteins and one that produced the mutant H5ts149 Ad pol. To investigate the structure-function relationships of both proteins, extracts from cells infected with the recombinant viruses were tested for in vitro complementation of the initiation and elongation steps in adenovirus DNA replication. The results were in accordance with those of earlier in vivo experiments with these insertion mutants and indicate that multiple regions of both proteins are essential for adenovirus DNA replication. The carboxyl termini of both pTP and Ad pol were shown to be essential for proper functioning of these proteins during initiation of adenovirus DNA replication. Three different DNA replication-negative pTP mutants were shown to have residual activity in the initiation assay, suggesting not only that pTP is required for initiation but also that it may play a role in DNA replication after the deoxycytidylation step. PMID- 8416373 TI - Release, uptake, and effects of extracellular human immunodeficiency virus type 1 Tat protein on cell growth and viral transactivation. AB - During acute human immunodeficiency virus type 1 (HIV-1) infection or after transfection of the tat gene, Tat protein is released into the cell culture supernatant. In this extracellular form, Tat stimulates both HIV-1 gene expression and the growth of cells derived from Kaposi's sarcoma (KS) lesions of HIV-1-infected individuals (AIDS-KS cells). Tat protein and its biological activities appear in the cell supernatants at the peak of Tat expression, when the rate of cell death is low (infection) or cell death is undetectable (transfection) and increased levels of cytoplasmic Tat are present. Tat containing supernatants stimulate maximal AIDS-KS cell growth but only low to moderate levels of HIV-1 gene expression. This is due to the different concentrations of exogenous Tat required for the two effects. The cell growth promoting effects of Tat peak at between 0.1 and 1 ng of purified recombinant protein per ml in the cell growth medium and do not increase with concentration. In contrast, both the detection of nuclear-localized Tat taken up by cells and the induction of HIV-1 gene expression or replication require higher Tat concentrations (> or = 100 ng/ml), and all increase linearly with increasing amounts of the exogenous protein. These data suggest that Tat can be released by a mechanism(s) other than cell death and that the cell growth-promoting activity and the virus-transactivating effect of extracellular Tat are mediated by different pathways. PMID- 8416374 TI - Mutational analysis of the p50 subunit of NF-kappa B and inhibition of NF-kappa B activity by trans-dominant p50 mutants. AB - The NF-kappa B family of DNA-binding proteins regulates the expression of many cellular and viral genes. Each of these proteins has an N-terminal region that is homologous to the c-Rel proto-oncogene product, and this Rel homology region defines both DNA binding and protein dimerization properties of the individual proteins. Most of the NF-kappa B family members have been shown to associate with themselves or with each other to form homodimers or heterodimers, and previous studies have shown that dimerization of NF-kappa B factors is necessary to provide a functional DNA binding domain. We have used site-directed mutagenesis to identify regions in the Rel homology domain of the p50/NF-kappa B protein that are important for DNA binding and protein dimerization. Our studies have identified mutations of p50 that interfere with DNA binding only and those that interfere with protein dimerization. Mutations of p50 which disrupt only DNA binding were still able to associate with other members of the NF-kappa B protein family. We demonstrate that such heterodimeric complexes inhibit transcriptional activation mediated in trans through a cis-acting kappa B motif; therefore, we have identified trans-dominant negative mutants of p50. PMID- 8416375 TI - Role of the large hepatitis B virus envelope protein in infectivity of the hepatitis delta virion. AB - The hepatitis delta virus (HDV) is coated with large (L), middle (M), and small (S) envelope proteins encoded by coinfecting hepatitis B virus (HBV). To study the role of the HBV envelope proteins in the assembly and infectivity of HDV, we produced three types of recombinant particles in Huh7 cells by transfection with HBV DNA and HDV cDNA: (i) particles with an envelope containing the S HBV envelope protein only, (ii) particles with an envelope containing S and M proteins, and (iii) particles with an envelope containing S, M, and L proteins. Although the resulting S-, SM-, and SML-HDV particles contained both hepatitis delta antigen and HDV RNA, only particles coated with all three envelope proteins (SML) showed evidence of infectivity in an in vitro culture system susceptible to HDV infection. We concluded that the L HBV envelope protein, and more specifically the pre-S1 domain, is important for infectivity of HDV particles and that the M protein, which has been reported to bear a site for binding to polymerized albumin in the pre-S2 domain, is not sufficient for infectivity. Our data also show that the helper HBV is not required for initiation of HDV infection. The mechanism by which the L protein may affect HDV infectivity is discussed herein. PMID- 8416376 TI - Characterization of human immunodeficiency virus type 1 integrase expressed in Escherichia coli and analysis of variants with amino-terminal mutations. AB - Replication of a retroviral genome depends upon integration of the viral DNA into a chromosome of the host cell. The integration reaction is mediated by integrase, a viral enzyme. Human immunodeficiency virus type 1 integrase was expressed in Escherichia coli and purified to near homogeneity. Optimum conditions for the integration and 3'-end-processing activities of integrase were characterized by using an in vitro assay with short, double-stranded oligonucleotide substrates. Mutants containing amino acid substitutions within the HHCC region, defined by phylogenetically conserved pairs of histidine and cysteine residues near the N terminus, were constructed and characterized by using three assays: 3'-end processing, integration, and the reverse of the integration reaction (or disintegration). Mutations in the conserved histidine and cysteine residues abolished both integration and processing activities. Weak activity in both assays was retained by two other mutants containing substitutions for less highly conserved amino acids in this region. All mutants retained activity in the disintegration assay, implying that the active site for DNA cleavage-ligation is not located in this domain and that the HHCC region is not the sole DNA-binding domain in the protein. However, the preferential impairment of processing and integration rather than disintegration by mutations in the HHCC region is consistent with a role for this domain in recognizing features of the viral DNA. This hypothesis is supported by the results of disintegration assays performed with altered substrates. The results support a model involving separate viral and target DNA-binding sites on integrase. PMID- 8416377 TI - Identification and characterization of the p35 gene of Bombyx mori nuclear polyhedrosis virus that prevents virus-induced apoptosis. AB - Nucleotide sequence analysis of the Bombyx mori nuclear polyhedrosis virus (BmNPV) genome revealed the existence of a gene homologous to the p35 gene of Autographa californica NPV (AcNPV), which has been shown to prevent virus-induced apoptosis. The BmNPV p35 gene showed 96.1% nucleotide and 89.6% predicted amino acid sequence identity to the AcNPV p35 gene. A mutant BmNPV (BmP35Z) lacking a functional p35 gene induced apoptosis-like cell degradation in infected BmN cells. However, unlike the p35-deleted AcNPV mutant (vAcAnh), BmP35Z replicated normally and produced polyhedral inclusion bodies. The patterns of protein synthesis and the percentages of viable BmN cells remaining following infection with either wild-type BmNPV or BmP35Z were nearly identical. BmP35Z also replicated in silkworm larvae without showing any apparent apoptotic response in infected hemocytes, fat body, or other tissues. Time to death of larvae infected with BmP35Z was similar to that for wild-type-infected larvae, and significant numbers of polyhedral inclusion bodies were produced. These results indicate that viral factors (or genes) other than p35 or host cell factors play a role in inducing, accelerating, or interfering with apoptotic processes. The evolution of baculovirus genomes is also discussed with reference to comparative analysis of the p35 and p94 gene sequences. The p94 gene is found immediately upstream of p35 in AcNPV; in BmNPV, however, the p94 gene was nearly completely missing, presumably because of large deletions in a BmNPV ancestor virus having a gene similar to the AcNPV p94 gene. PMID- 8416378 TI - Immunodominance of the VP4 neutralization protein of rotavirus in protective natural infections of young children. AB - Natural infection by very similar strains of rotavirus during the 1988-1989 rotavirus season in Cincinnati, Ohio, provided complete protection of young children against subsequent rotavirus illnesses for a period of at least 2 years. Using this limited strain variability, we characterized the association between the titers of antibody to either the VP4 or the VP7 neutralization protein and protection against subsequent rotavirus disease. This was done by using reassortants that contained only one of the two rotavirus neutralization proteins of 89-12, a culture-adapted isolate representative of the protective rotavirus strains. The other neutralization protein in these reassortants was derived from a heterologous rotavirus (WC3 or EDIM) to which the infected subjects made little or no neutralizing antibody (titers, < or = 20). The geometric mean titer (GMT) of antibody to 89-12 in convalescent-phase sera from the 21 subjects analyzed was 2,323. The GMT of antibody to a reassortant (strain WC-4) that contained the VP7 protein of 89-12 and VP4 of WC3 was 387. In contrast, the GMT of antibody to a reassortant (strain EDIM-7) that contained the VP4 protein of 89-12 and the VP7 protein of EDIM was 1,078. Thus, the major neutralization response was directed against VP4 rather than VP7, a finding that has important implications for development of appropriate rotavirus vaccines. PMID- 8416379 TI - Identification of specific adenovirus E1A N-terminal residues critical to the binding of cellular proteins and to the control of cell growth. AB - Adenovirus early region 1A (E1A) oncogene-encoded sequences essential for transformation- and cell growth-regulating activities are localized at the N terminus and in regions of highly conserved amino acid sequence designated conserved regions 1 and 2. These regions interact to form the binding sites for two classes of cellular proteins: those, such as the retinoblastoma gene product, whose association with the E1A products is specifically dependent on region 2, and another class which so far is known to include only a large cellular DNA binding protein, p300, whose association with the E1A products is specifically dependent on the N-terminal region. Association between the E1A products and either class of cellular proteins can be disrupted by mutations in conserved region 1. While region 2 has been studied intensively, very little is known so far concerning the nature of the essential residues in the N-terminal region, or about the manner in which conserved region 1 participates in the binding of two distinct sets of cellular proteins. A combination of site-directed point mutagenesis and monoclonal antibody competition experiments reported here suggests that p300 binding is dependent on specific, conserved residues in the N terminus, including positively charged residues at positions 2 and 3 of the E1A proteins, and that p300 and pRB bind to distinct, nonoverlapping subregions within conserved region 1. The availability of precise point mutations disrupting p300 binding supports previous data linking p300 with cell cycle control and enhancer function. PMID- 8416380 TI - Borna disease virus in mice: host-specific differences in disease expression. AB - We developed a mouse model of Borna disease to facilitate immunopathogenesis research by adaptation of Borna disease virus to mice through serial passage in mouse brain tissue. Borna disease virus replication, antibody production, inflammation, and Borna disease expression in several different strains of mice were examined. PMID- 8416381 TI - Transcriptional mapping of a late gene coding for the p12 attachment protein of African swine fever virus. AB - The transcriptional characterization of the gene coding for the p12 attachment protein of the African swine fever virus is presented. The results obtained have been used to generate the first detailed transcriptional map of an African swine fever virus late gene. Novel experimental evidence indicating the existence of major differences between the mechanisms controlling the transcription of late genes in African swine fever virus and poxviruses is provided. PMID- 8416382 TI - The cytoplasmic domain of the human T-cell leukemia virus type I envelope can modulate envelope functions in a cell type-dependent manner. AB - C-terminal truncations of the human T-cell leukemia virus type I envelope affected the intracellular maturation and syncytium formation in a cell type dependent manner. The intracytoplasmic domain appears dispensable for syncytium formation, but its truncation can modulate the envelope functionality in some cell types. PMID- 8416383 TI - Successful replication of parvovirus B19 in the human megakaryocytic leukemia cell line MB-02. AB - The pathogenic human parvovirus B19 has been shown to undergo productive replication in the erythroid lineage in primary normal human hematopoietic progenitor cells. However, none of the established erythroleukemia cell lines has allowed B19 virus replication in vitro. The remarkable erythroid tissue tropism of B19 virus was evaluated with a human megakaryocytic leukemia cell line, MB-02, which is dependent on the growth factor granulocyte-macrophage colony-stimulating factor but can be induced to undergo erythroid differentiation following treatment with erythropoietin (Epo). Whereas these cells did not support B19 virus DNA replication in the presence of granulocyte-macrophage colony stimulating factor alone, active viral DNA replication was observed if the cells were exposed to Epo for 5 to 10 days prior to B19 virus infection, as detected by the presence of the characteristic B19 virus DNA replicative intermediates on Southern blots. No replication occurred if the cells were treated with Epo for 3 days or less. In addition, complete expression of the B19 virus genome also occurred in Epo-treated MB-02 cells, as detected by Northern blot analysis. B19 progeny virions were released into culture supernatants that were biologically active in secondary infection of normal human bone marrow cells. The availability of the only homogeneous permanent cell line in which induction of erythroid differentiation leads to a permissive state for B19 virus replication in vitro promises to yield new and useful information on the molecular basis of the erythroid tissue tropism as well as parvovirus B19-induced pathogenesis. PMID- 8416384 TI - Immune response of rhesus macaques to recombinant simian immunodeficiency virus gp130 does not protect from challenge infection. AB - Simian immunodeficiency virus (SIV) infection of rhesus macaques is a model for human immunodeficiency virus (HIV) infection in humans. Inactivated and modified live whole-virus vaccines have provided limited protective immunity against SIV in rhesus macaques. Because of safety concerns in the use of inactivated and live whole-virus vaccines, we evaluated the protective immunity of vaccinia virus recombinants expressing the surface glycoprotein (gp130) of SIVmac and subunit preparations of gp130 expressed in mammalian cells (CHO). Three groups of animals were immunized with recombinant SIV gp130. The first group received SIV gp130 purified from genetically engineered CHO cells (cSIVgp130), the second group was vaccinated with recombinant vaccinia virus expressing SIVmac gp130 (vSIVgp130), and the third group was first primed with vSIVgp130 and then given a booster immunization with cSIVgp130. Although anti-gp130 binding antibodies were elicited in all three groups, neutralizing antibodies were transient or undetectable. None of the immunized animals resisted intravenous challenge with a low dose of cell free virus. However, the group primed with vSIVgp130 and then boosted with cSIVgp130 had the lowest antigen load (p27) compared with the other groups. The results of these studies suggest that immunization of humans with HIV type 1 surface glycoprotein may not provide protective immunity against virus infection. PMID- 8416385 TI - Glycosylation is necessary for the correct folding of human immunodeficiency virus gp120 in CD4 binding. AB - Conflicting results have been reported regarding the role of carbohydrate on human immunodeficiency virus (HIV) envelope glycoprotein gp120 in CD4 receptor binding. Glycosylated, deglycosylated, and nonglycosylated forms of HIV type 1 (HIV-1) and HIV-2 gp120s were used to examine CD4 receptor-binding activity. Nonglycosylated forms of gp120 generated either by deletion of the signal sequence of HIV-1 gp120 or by synthesis in the presence of tunicamycin failed to bind to CD4. In contrast, highly mannosylated gp120 bound to soluble CD4 molecules well. Enzymatic removal of carbohydrate chains from glycosylated gp120 by endoglycosidase H or an endoglycosidase F/N glycanase mixture had no effect on the ability of gp120 to bind CD4. An experiment which measured the ability of gp120 to bind to CD4 as an assay of the proper conformation of gp120 showed that carbohydrate chains on gp120 are not required for the interaction between gp120 and CD4 but that N-linked glycosylation is essential for generation of the proper conformation of gp120 to provide a CD4-binding site. PMID- 8416386 TI - Theiler's virus infection of beta 2-microglobulin-deficient mice. AB - Theiler's virus, a murine picornavirus, persists in the central nervous systems of susceptible mice and induces a chronic demyelinating disease. Susceptibility or resistance to this disease is controlled in part by the H2-D locus of the major histocompatibility complex (MHC). For this reason, it has been proposed that CD8+ class I-restricted cytotoxic T cells play a main role in the pathogenesis of this viral infection. We recently reported the existence of anti virus CD8+ cytotoxic T cells in the course of Theiler's virus infection. In the present study, we examined the role of these effector cells in mice in which the beta 2-microglobulin gene had been disrupted. These mice fail to express class I MHC molecules and therefore lack CD8+ T cells. The mice are derived from a C57BL/6 x 129/Ola cross and are H-2b, a haplotype associated with resistance to Theiler's virus infection. beta 2-Microglobulin-deficient mice (beta 2m-/-mice) failed to clear the virus, developed demyelination, and, interestingly, did not succumb to early infection. These results demonstrate that CD8+ T cells are required to clear Theiler's virus infection. In contrast with a current hypothesis, they also demonstrate that CD8+ T cells are not major mediators of the demyelinating disease. PMID- 8416387 TI - A mutant CHO-K1 strain with resistance to Pseudomonas exotoxin A is unable to process the precursor fusion glycoprotein of Newcastle disease virus. AB - RPE.40, a mutant strain of CHO-K1 cells isolated for resistance to Pseudomonas exotoxin A and cross-resistant to alphaviruses, is also highly resistant to virulent strains of Newcastle disease virus. The resistance of RPE.40 cells to Newcastle disease virus results from the failure to cleave the viral envelope precursor glycoprotein Fo to fusion glycoprotein F1 at the consensus sequence (Lys/Arg)-Arg-Gln-(Lys/Arg)-Arg. PMID- 8416388 TI - Multipartite nature of potato virus X. AB - Potato virus X (PVX) was among the first viruses to be purified. Nonetheless, properties of the purified virus remain contentious. The literature has been heavily influenced by the concept of a virus as a monopartite entity. Despite the fact that electron micrographs invariably show large proportions of shorter virus particles, the latter are universally ignored. Seven distinct classes of particle lengths were detected. Seven RNA species of approximate sizes 6.4, 3.6, 3.0, 2.1, 1.8, 1.4, and 0.9 kb were extracted from these purified virus preparations. This study shows clearly that shorter PVX particles are not breakage products and indicates that they may reflect fundamental properties of the genome strategy. Furthermore, other potexviruses have been found to contain many shorter particles, and the level of these particles is stable during purification. PVX is generally believed to consist of particles of single length even though the literature does not confirm this conclusion. The notion of a single particle length appears to reflect historical concepts of what a virus should be rather than what PVX is. This report considers whether shorter rods present in virus preparations of PVX are distinctive products of infection. The problem addressed is significant because if affects conclusions concerning the mechanisms of PVX biosynthesis and replication. PMID- 8416389 TI - Cell fusion induced by the murine leukemia virus envelope glycoprotein. AB - To determine whether ecotropic murine leukemia virus (MuLV) envelope glycoproteins are sufficient to cause cell-to-cell fusion when expressed in the absence of virus production, we used an ecotropic MuLV, AKV, to construct env expression vectors that lack the gag and pol genes. The rat cell line XC, which undergoes cell-to-cell fusion upon infection with ecotropic MuLV, was transfected with wild-type env expression vectors, and high levels of syncytium formation resulted. Transfection of the murine cell line NIH 3T3 with expression vectors containing the wild-type or mutated env region did not result in syncytium formation. Immunoprecipitation analysis of the envelope glycoproteins expressed in NIH 3T3 and XC cells showed that the mature surface glycoprotein expressed in XC cells was of a much lower apparent molecular weight than that expressed in NIH 3T3 cells. Further characterization showed that most if not all of this difference was the result of differences in glycosylation. Finally, site-directed mutagenesis was used to introduce several conservative and nonconservative changes into the amino-terminal region of the transmembrane protein. Analysis of the effect of these mutations confirmed that this region is a fusion domain. PMID- 8416390 TI - Somalia operation just one of many demands on US military medicine. PMID- 8416391 TI - Khat abuse fuels Somali conflict, drains economy. PMID- 8416392 TI - PolioPlus: poliomyelitis eradication by year 2005. PMID- 8416393 TI - Delayed trial of HIV immune globulin to protect infants of infected mothers is likely to resume. PMID- 8416394 TI - From the Centers for Disease Control and Prevention. Population-based mortality assessment: Baidoa and Afgoi, Somalia, 1992. PMID- 8416395 TI - From the Centers for Disease Control and Prevention. Poliomyelitis outbreak- Netherlands, 1992. PMID- 8416396 TI - From the Centers for Disease Control and Prevention. Knowledge of the purpose of community water fluoridation--United States, 1990. PMID- 8416397 TI - Patient mix and utilization of resources. PMID- 8416398 TI - Student loan deferment. PMID- 8416399 TI - Qualifications for administrative medicine: is an MD enough? PMID- 8416400 TI - Laser vs cryotherapy for CIN and a LEEP of faith. PMID- 8416401 TI - The failure of death certificates to record the performance of autopsies. PMID- 8416402 TI - Transdermal medication exchange: the case of the light-headed lover. PMID- 8416404 TI - Measuring health care quality. PMID- 8416403 TI - AIDS and confidentiality: making exemptions to encourage research. PMID- 8416405 TI - A piece of my mind. Given the choice. PMID- 8416406 TI - Adult immunization with acellular pertussis vaccine. AB - OBJECTIVE: To evaluate the safety and immunogenicity in adults of several different concentrations of an acellular pertussis vaccine. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: Medical center immunization clinic. PARTICIPANTS: One hundred eighteen healthy adult volunteers. INTERVENTIONS: Participants received standard adult tetanus-diphtheria vaccine alone or combined with full-strength, half-strength, or quarter-strength concentrations of a currently licensed acellular pertussis vaccine used for booster doses in young children. Full-strength vaccine contained 40 micrograms of pertussis proteins, consisting of 86% filamentous hemagglutinin, 8% pertussis toxin, 4% 69-kd outer-membrane protein, and 2% agglutinogens. MAIN OUTCOME MEASURES: Local and systemic reactions were assessed for 14 days after vaccination. Serum samples for antibody assay were obtained before, 1 month after, and 1 year after immunization. RESULTS: Adverse reactions were few and minor and did not differ in frequency or severity among the four study groups. The groups receiving acellular pertussis vaccine showed strong antibody responses to pertussis antigens, which did not significantly differ by concentration of vaccine. After 1 year, levels of antibody to pertussis had declined by approximately 50% but remained substantially higher than preimmunization levels. The four groups did not differ in antibody responses to tetanus or diphtheria toxoids. CONCLUSIONS: Routine reimmunization of adults with a vaccine containing acellular pertussis antigens in addition to diphtheria and tetanus toxoids can substantially enhance pertussis antibody levels without an increase in adverse reactions or diminution in response to the diphtheria and tetanus components. Such a program might materially reduce respiratory illness among both adults and children. PMID- 8416407 TI - Disease-specific survival following routine prostate cancer screening by digital rectal examination. AB - OBJECTIVE: To assess prostate cancer mortality in men undergoing routine screening by routine digital rectal examination. DESIGN: Cohort study with a median follow-up period of 75 months. SETTING: Population consisted of volunteers at a university clinic and men in an institutional health maintenance clinic. PATIENTS: Fifty-six men with a mean age of 65 years (range, 52 to 79 years) diagnosed with prostate cancer. INTERVENTIONS: Patients treated initially by observation, external or interstitial radiotherapy, radical prostatectomy, hormone therapy, or combination. MAIN OUTCOME MEASURES: Kaplan-Meier analysis of time to local progression, distant metastases, death from all causes, and death from prostate cancer. Mantel-Haenszel log-rank statistic was used to compare outcome in men diagnosed on initial examination with those diagnosed on subsequent examinations. RESULTS: Clinically localized prostate cancer was diagnosed in 73% during an initial examination and 83% on subsequent examinations and (P.35). Grade distribution of tumors was similar in both groups. Overall 5 and 10 year survival of all cancer patients was 85% and 67%, respectively. Death from prostate cancer was 8% (3/38) in men diagnosed on initial examination and 33% (6/18) during subsequent examinations. Five- and 10-year disease-specific survival was 97% and 86%, respectively, for men diagnosed during the first rectal examination compared with only 81% and 57%, respectively, for men diagnosed on subsequent rectal examinations (P = .02). CONCLUSION: Routine screening for prostate cancer by annual digital rectal examination alone may be insufficiently frequent and/or sensitive to prevent significant mortality from this disease [corrected]. PMID- 8416408 TI - Predictors and prognosis of inability to get up after falls among elderly persons. AB - OBJECTIVE: To identify the predictors and prognosis associated with inability to get up after falling. DESIGN: Cohort study with a mean 21-month follow-up. SETTING: General community. SUBJECTS: 1103 New Haven, Conn, residents aged 72 years and older who were able to follow simple commands and walk unassisted. MAIN OUTCOME MEASURES: Self-reported inability to get up without help after falls not resulting in serious injury; activity restriction and hospitalization after a fall; death; and placement in a nursing home. RESULTS: Inability to get up without help was reported after 220 of 596 non-injurious falls. Of 313 non injured fallers, 148 (47%) reported inability to get up after at least one fall. Compared with non-fallers, the risk factors independently associated with inability to get up included the following: an age of at least 80 years (adjusted relative risk [RR], 1.6; 95% confidence interval [CI], 1.2 to 2.1); depression (RR, 1.5; CI, 1.1 to 2.0); and poor balance and gait (RR, 2.0; CI, 1.5 to 2.7). Previous stroke (RR, 1.6; CI, 1.0 to 2.4) and sedative use (RR, 1.5; CI, 0.9 to 2.2) did not achieve significance. Among fallers, older age and poor balance and gait were associated marginally with inability to get up. Compared with fallers who were able to get up, fallers who were unable to get up were more likely to suffer lasting decline in activities of daily living (35% vs 26%). Fallers who were unable to get up were more likely to die, to be hospitalized, and to suffer a decline in activities of daily living for at least 3 days, and were less likely to be placed in a nursing home than were fallers who were able to get up, but these trends were not statistically significant. CONCLUSIONS: The risk factors for inability to get up were similar to those for falling, although certain factors imparted a particular risk of inability to get up without help. The frequency of inability to get up and the short- and long-term morbidity associated with this inability suggest the need for preventive and treatment efforts. PMID- 8416409 TI - Diagnosis and treatment of Clostridium difficile colitis. AB - Pseudomembranous colitis associated with antibiotic therapy is almost always due to an overgrowth of Clostridium difficile. If untreated, pseudomembranous colitis can lead to severe diarrhea, hypovolemic shock, toxic dilatation of the colon, cecal perforation, hemorrhage, and death. However, C difficile-associated colitis can mimic the more common "benign" antibiotic-associated diarrhea that is not caused by C difficile. An algorithm for diagnosis management of hospitalized patients with antibiotic diarrhea and C difficile colitis is presented in this review. Diagnosis depends on sigmoidoscopy and/or stool tests for C difficile toxins in all patients with antibiotic-associated diarrhea. If the results of these tests are positive, either metronidazole or vancomycin is recommended for treatment of mild illness, and vancomycin is recommended for treatment of severe illness. Oral therapy is always preferred because it is more reliable. In patients with recurrent or relapsing colitis, treatment with either metronidazole or vancomycin is effective for that episode, but novel approaches, such as the oral or rectal introduction of competing nonpathogenic organisms, may prove to be more successful in prevention of relapses. PMID- 8416410 TI - Cats and toxoplasmosis risk in HIV-infected adults. AB - OBJECTIVE: To establish the prevalence and incidence of toxoplasmosis in an adult human immunodeficiency virus (HIV) population, and to determine if cat ownership contributes to the risk of toxoplasmosis. DESIGN: Retrospective record and laboratory review, coupled with a patient survey. SETTING: A tertiary-care military hospital HIV program. PATIENTS: A total of 723 HIV-infected adults, all former or current US military personnel. MAIN OUTCOME MEASURES: Prevalence and incidence of serologic evidence of toxoplasmosis infection. RESULTS: A total of 723 HIV-infected patients had serial Toxoplasma IgG antibody determinations. Seventy patients (9.7%) were positive on their initial screen; the seronegative patients were tested annually for 1 to 5 years (mean duration of follow-up, 2.1 years). Only 13 patients (2.0%) who were initially seronegative acquired antibodies to Toxoplasma gondii. None of the patients who seroconverted developed clinical disease. A pet history was available on 12 of 13 patients who seroconverted; only one (8.3%) had owned or lived in a household with a cat during the period of seroconversion. The calculated attributable risk of cat ownership/exposure for toxoplasmosis seroconversion in this population is -2.9 per 100 persons annually. CONCLUSION: Toxoplasma antibody seroconversion in an adult HIV-infected population is unusual and appears unrelated to cat ownership or exposure. PMID- 8416411 TI - Immunotoxin therapy for cancer. PMID- 8416412 TI - Mandatory parental consent to abortion. Council on Ethical and Judicial Affairs, American Medical Association. AB - This report analyzes the ethical issues raised by requirements that parents be involved when minors seek an abortion. Parents are generally supportive and understanding and can provide helpful guidance to their children. In some cases, however, parents may respond abusively to the knowledge that their minor child is pregnant or is considering an abortion. In addition, privacy in matters of health care is a profound need of minors as well as adults. Accordingly, the Council concludes that, while minors should be encouraged to discuss their pregnancy with their parents and other adults, minors should not be required to involve their parents before deciding whether to undergo an abortion. PMID- 8416413 TI - The effect of providing health coverage to poor uninsured pregnant women in Massachusetts. AB - OBJECTIVES: There has been substantial policy interest in whether the provision of health coverage to poor uninsured pregnant women affects access to prenatal care and birth outcomes. We therefore examined whether the statewide provision of health coverage to uninsured low-income pregnant women affects access to prenatal care and infant birth outcomes. DESIGN: Natural experiment. PATIENTS: All in hospital, single-gestation live births in 1984 (N = 57,257) and 1987 (N = 64,346). INTERVENTION: In 1985, Massachusetts instituted Healthy Start, a program providing health coverage to uninsured pregnant women with incomes below 185% of the federal poverty level. MAIN OUTCOME MEASURES: Rates of satisfactory prenatal care, care initiated before the third trimester, and adverse infant outcome for uninsured women and for two concurrent control groups, women with Medicaid, and women with private insurance. We calculated the difference in rates between the uninsured and each concurrent control. To assess the effect of the program, we examined the change in these interpayer differences in rates between 1984 and 1987. MAIN RESULTS: Between 1984 and 1987, the rate of satisfactory prenatal care declined from 96.4% to 93.8% for all women in Massachusetts (P < .001). There was no statewide change in the overall incidence of adverse birth outcome (6.6% in both years). In 1984, uninsured women were less likely than privately insured women to receive satisfactory prenatal care (90.5% and 98.1%, respectively; interpayer difference, -7.6%) and to initiate care before the third trimester (94.2% and 99.1%; interpayer difference, -4.9%), and were more likely to suffer an adverse birth outcome (7.1% and 5.8%; interpayer difference, 1.3%). Between 1984 and 1987, there were no statistically significant changes in the interpayer differences in rates for any of the outcome measures relative to either control group. CONCLUSIONS: Our findings suggest that access to prenatal care may have declined for all women in Massachusetts between 1984 and 1987. In the setting of this statewide decline in access, the expansion of health coverage to uninsured low-income pregnant women was not associated with an improvement in access to prenatal care or birth outcomes. PMID- 8416414 TI - AIDS-associated primary central nervous system lymphoma. Oncology Core Committee, AIDS Clinical Trials Group. PMID- 8416415 TI - Pertussis and pertussis vaccines in adults. PMID- 8416416 TI - The next era for prostate cancer. Controlled clinical trials. PMID- 8416417 TI - Effects of continuous infusion of nitroglycerin on pulmonary hemodynamics, lung lymph balance, and prostanoid products in the response to endotoxin in awake sheep. AB - We examined the effects of continuous intravenous infusion of nitroglycerin (NTG) on lung dysfunction induced by endotoxemia in awake sheep chronically instrumented with lung lymph fistula. We measured the responses of hemodynamics, lung lymph balance, and thromboxane (Tx)B2 and 6-keto-prostaglandin (PG) F1 alpha levels in plasma and lung lymph to endotoxin administration (1 microgram/kg, intravenously [IV], over 30 min) with and without continuous infusion of NTG (1 microgram/kg/min). Continuous infusion of NTG alone (n = 5) over 5 hr did not significantly alter systemic, pulmonary hemodynamics, and/or lung lymph fluid filtration. Infusion of endotoxin alone (n = 7) caused remarkable increases in pulmonary artery pressure (Ppa) and lung lymph flow (Qlym) in the early phase. Continuous infusion of NTG (n = 6) significantly prevented the early increases in Ppa and Qlym after endotoxin. The increased values of TxB2 and 6-keto-PGF1 alpha in both plasma and lung lymph after endotoxemia showed the same increases in groups with and without NTG. These findings suggest that the reduction of pulmonary artery pressure induced by NTG decreased the filtration of fluid into the lungs associated with endotoxemia in sheep, and that the mechanism of vasodilating action of NTG is not due to modifications of constrictive-dilated cyclo-oxygenase products of arachidonate, such as TxA2 and PGI2. PMID- 8416418 TI - [123I]HIPDM pulmonary imaging demonstrates elastase-induced pulmonary emphysema. AB - Because more than 90 percent of [123I]hydroxyiodobenzyl-propanediamine (HIPDM) is localized in the lung after intravenous injection, the radiopharmaceutical has been proposed as a lung imaging agent. Its potential usefulness for the detection of pulmonary emphysema was evaluated in an animal model of elastase-induced emphysema along with [99mTc]MAA lung perfusion imaging. To induce lung emphysema, Long-Evans rats (200-250 g) were given 400 IU/Kg elastase intratracheally under ether anesthesia. Four weeks after elastase treatment, 15 treated and 15 nontreated rats were paired and simultaneously imaged under a scintillation camera immediately following intravenous injection of 0.25-0.3 mCi[99mTc]MAA. The procedure was repeated 48 hr later using 0.25-0.3 mCi[123I]HIPDM. Activity in the region of interest (ROI) over the lungs was recorded after the injections. Total counts per ROI from each rat were measured and normalized by lung volume. The normalized lung activity ratio of treated/nontreated rats was computed. The mean ratios of HIPDM and MAA were 0.847 and 0.802, respectively. The significant decrease in uptake of both HIPDM (p < 0.021) and of MAA (p < 0.025) in the elastase-treated lungs indicates decrease in functioning vascular endothelium and decrease in number of pulmonary capillary vessels, respectively, reflecting damage in capillaries and small arterioles. The decrease in treated/nontreated ratios lung is consistent with a significant alteration in pulmonary function and a significant increase in mean linear intercept (p < 0.005) in treated lung. Since the imaging reflects pulmonary endothelial receptor function, [123I]HIPDM lung imaging may serve as an alternative diagnostic modality for pulmonary emphysema. PMID- 8416419 TI - Influence of lung volume hysteresis on collateral resistance in intact dogs. AB - Collateral resistance (Rcoll) is highly lung volume dependent. We studied 12 dogs in an attempt to evaluate the influence of hysteresis on this volume dependency. Rcoll measurements were obtained at baseline and at different lung volumes as modified by the application of negative or positive extrathoracic pressure (ETP) in an iron lung. ETP was modified in 5 cmH2O steps from +20 to -25 cmH2O) and back to +20 cmH2O on the first day (cycle 1), and, in 4 of these dogs, from -25 to +20 cmH2O and back to -25 cmH2O on a second day (cycle 2). The behavior of Rcoll for both cycles was the same in all dogs, varying inversely with the changes in end-expiratory lung volume (EELV). All dogs presented a similar pattern: during cycle 1, for a same lung volume, Rcoll was much higher during the deflation limb than during the inflation limb. For cycle 2, Rcoll at a given lung volume was similar whether measured during the inflation or deflation limb. Changes in EELV were higher for any given negative ETP during the deflation than during the inflation limb of the pressure cycles (e.g., for the 4 dogs who had both cycles, at -15 cmH2O ETP of cycle 2 the increase in EELV was 1167 +/- 121 ml during deflation and 525 +/- 102 ml during the subsequent inflation [mean +/- SEM]). In conclusion, the effects of hysteresis on Rcoll and on lung volumes are similar, except at low lung volumes during the deflation limb of cycle 2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416420 TI - Chest wall configuration assessed at total lung capacity during acute asthma and after recovery. AB - During acute asthma reversible increase in lung compliance and total lung capacity (TLC) have been observed. The magnitude of the increase in TLC, however, was found to be relatively small when TLC was measured radiographically. We wondered if structural distortion of the respiratory bellows develops during acute asthma attacks, accounting for the limited increase in TLC. We analyzed the chest wall dimensions using chest roentgenograms obtained in 32 patients who had previously participated in the study of radiographic evaluation of TLC. We found that the dimensional changes accompanying the small increase in TLC during acute asthma were nonuniform, consisting of mainly an increase in lung height (0.45 +/- 0.15 cm) with limited coordinated expansion of other dimensions and a small but significant distortional contraction of lung width at the lowest portion of the rib cage (-0.25 +/- 0.12 cm). These findings were interpreted to mean that in response to the decrease in elastic lung recoil (internal load) occurring during acute asthma, the diaphragm makes limited but effective further caudad descent without provoking serious structural distortion and that rib cage muscles, working at mechanical disadvantage at high lung volume, act largely as fixators. PMID- 8416421 TI - Brief report: recognition of acute myocarditis masquerading as acute myocardial infarction. PMID- 8416422 TI - Prenatal diagnosis. PMID- 8416423 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 2-1993. A 72-year-old woman with a coagulopathy and bilateral thigh masses. PMID- 8416424 TI - Radiation and pituitary dysfunction. PMID- 8416425 TI - Prevention of Lyme disease after tick bites. PMID- 8416426 TI - Prevention of Lyme disease after tick bites. PMID- 8416427 TI - Prevention of Lyme disease after tick bites. PMID- 8416428 TI - Prevention of Lyme disease after tick bites. PMID- 8416429 TI - Prevention of Lyme disease after tick bites. PMID- 8416430 TI - Prevention of Lyme disease after tick bites. PMID- 8416431 TI - Prevention of Lyme disease after tick bites. PMID- 8416432 TI - Possibly disappointing results of treatment with gemfibrozil. PMID- 8416433 TI - Tamoxifen and chemotherapy for refractory metastatic malignant melanoma. PMID- 8416434 TI - Correction: location of bcl-2 oncogene. PMID- 8416435 TI - Normal reference laboratory values? PMID- 8416436 TI - Endogenous retroviruses in xenografts. PMID- 8416437 TI - The marketplace in health care reform. The demographic limitations of managed competition. AB - BACKGROUND: The theory of managed competition holds that the quality and economy of health care delivery will improve if independent provider groups compete for consumers. In sparsely populated areas where relatively few providers are required, however, it is not feasible to divide the provider community into competing groups. We examined the demographic features of health markets in the United States to see what proportion of the population lives in areas that might successfully support managed competition. METHODS: The ratios of physicians to enrollees in large staff-model health maintenance organizations were determined as an indicator of the staffing needs of an efficient health plan. These ratios were used to estimate the populations necessary to support health organizations with various ranges of specialty services. Metropolitan areas with populations large enough to support managed competition were identified. RESULTS: We estimated that a health care services market with a population of 1.2 million could support three fully independent plans. A population of 360,000 could support three plans that independently provided most acute care hospital services, but the plans would need to share hospital facilities and contract for tertiary services. A population of 180,000 could support three plans that provided primary care and many basic specialty services but that shared inpatient cardiology and urology services. Health markets with populations greater than 180,000 would include 71 percent of the U.S. population; those with populations greater than 360,000, 63 percent; and those with populations greater than 1.2 million, 42 percent. CONCLUSIONS: Reform of the U.S. health care system through expansion of managed competition is feasible in medium-sized or large metropolitan areas. Smaller metropolitan areas and rural areas would require alternative forms of organization and regulation of health care providers in order to improve quality and economy. PMID- 8416438 TI - Hypothalamic-pituitary dysfunction after radiation for brain tumors. AB - BACKGROUND: Patients with brain tumors who are treated with radiation frequently have growth hormone deficiency, but other neuroendocrine abnormalities are presumed to be uncommon. METHODS: We studied endocrine function in 32 patients (age, 6 to 65 years) 2 to 13 years after they had received cranial radiotherapy for brain tumors. The doses of radiation to the hypothalamic-pituitary region ranged from 3960 to 7020 rad (39.6 to 70.2 Gy). Nine patients also received 1800 to 3960 rad (18.0 to 39.6 Gy) to the craniospinal axis. Serum concentrations of thyroid, gonadal, and pituitary hormones were measured at base line and after stimulation. RESULTS: Nine patients (28 percent) had symptoms of thyroid deficiency, and 20 patients (62 percent) had low serum total or free thyroxine or total triiodothyronine concentrations. Of the 23 patients treated only with cranial radiation, 15 (65 percent) had hypothalamic or pituitary hypothyroidism. Of the nine patients who also received spinal (and thus direct thyroid) radiation, three (33 percent) had evidence of primary thyroid injury. Seven of the 10 postpubertal, premenopausal women (70 percent) had oligomenorrhea, and 5 (50 percent) had low serum estradiol concentrations. Three of the 10 men (30 percent) had low serum testosterone concentrations. Overall, 14 of the 23 postpubertal patients (61 percent) had evidence of hypogonadism. Mild hyperprolactinemia was present in 50 percent of the patients. Responses to stimulation with corticotropin-releasing hormone and corticotropin were normal in all patients except one, who had panhypothalamic dysfunction. However, serum 11 deoxycortisol responses to the administration of metyrapone were low in 11 of the 31 patients (35 percent) tested. Three of the 32 patients, (9 percent) had no endocrine abnormalities, 9 (28 percent) had an abnormal result on tests of thyroid, gonadal, prolactin, or adrenal function, 8 (25 percent) had abnormalities in two axes, 8 (25 percent) in three axes, and 4 (12 percent) in all four axes. CONCLUSIONS: Cranial radiotherapy in children and adults with brain tumors frequently causes abnormal hypothalamic-pituitary function. The most frequent changes are hypothyroidism and gonadal dysfunction, although subtle abnormalities in adrenal function may also be present. PMID- 8416439 TI - Microsporidia infection in patients with the human immunodeficiency virus and unexplained cholangitis. AB - BACKGROUND: Cholangitis in patients with the acquired immunodeficiency syndrome (AIDS) is usually associated with opportunistic infections by cryptosporidium species or cytomegalovirus, but in about a third of cases no opportunistic agent is identified. We suspected some of these cases of biliary disease might be explained by infection with the microsporidia species Enterocytozoon bieneusi, an obligate intracellular protozoan that causes chronic diarrhea in patients infected with the human immunodeficiency virus (HIV). METHODS: We studied eight HIV-infected homosexual men (in either group IV of the classification of the Centers for Disease Control and Prevention or group II, with a CD4 cell count of < or = 10 per cubic millimeter) who were referred because of cholangitis for which no causative agent had been found by standard tests. All the patients underwent abdominal ultrasonography and endoscopic ultrasonography or endoscopic retrograde cholangiopancreatography with collection of bile from the common bile duct. One patient had transhepatic biliary catheterization, and two others had cholecystectomy. Bile samples, duodenal- and liver-biopsy specimens, and gallbladder tissue were studied by light and electron microscopy. RESULTS: All eight patients with unexplained AIDS-related cholangitis had biliary microsporidosis. Intraepithelial E. bieneusi spores (1 to 2 microns) and supranuclear plasmodia (3 to 8 microns) were identified in the six duodenal biopsy specimens. May-Grunwald-Giemsa staining of bile samples revealed free forms of microsporidia in all eight patients, and the presence of E. bieneusi was confirmed by electron microscopy. E. bieneusi was also identified in ductal biliary cells on a liver biopsy, in one common-bile-duct smear, and in gallbladder epithelium (in two patients). Four patients were found to have associated but previously undetected biliary or duodenal cryptosporidiosis, whereas another had biliary infection associated with cytomegalovirus. CONCLUSIONS: Infection of the biliary tract with E. bieneusi is associated with and may be a cause of AIDS-related cholangitis. PMID- 8416440 TI - Domestic violence: let our voices be heard. PMID- 8416441 TI - The prevention of bone loss in young women treated with GnRH analogues with "add back" estrogen therapy. AB - OBJECTIVE: To determine whether the addition of a low dose of oral estrogen replacement therapy (ERT) taken daily can prevent the bone loss associated with continuous GnRH analogue use. METHODS: In a double-blind, placebo-controlled study, 60 women aged 21-45 years were randomized to one of three treatment groups: placebo implant every 4 weeks plus placebo ERT tablets daily, Zoladex (goserelin 3.6 mg) implant every 4 weeks plus placebo ERT tablets daily, or Zoladex (3.6 mg) implant every 4 weeks plus estradiol valerate, 2 mg/day, with norethisterone 5 mg from days 22-28. A dual x-ray bone density scan was performed before treatment and again after six treatment cycles. The percentage bone change with respect to the initial bone density was calculated. RESULTS: There was a significant loss of bone density at both the lumbar spine and proximal femur in the group receiving Zoladex plus placebo after 6 months compared with both pre treatment values and with the group receiving placebo plus placebo. The addition of estrogen "add-back" therapy to GnRH analogue treatment (Zoladex plus ERT) resulted in no significant change in bone density compared with either pre treatment values or the group receiving placebo plus placebo. The study had a dropout rate of 32%. CONCLUSION: The addition of "add-back" estrogen therapy to continuous GnRH analogue use can prevent bone loss. PMID- 8416442 TI - Management of severe ovarian hyperstimulation syndrome by ascitic fluid aspiration and intensive intravenous fluid therapy. AB - OBJECTIVE: To assess the value of intensive intravenous (IV) fluid therapy and ascitic fluid aspiration in the management of severe ovarian hyperstimulation syndrome. METHODS: Forty-two women with severe ovarian hyperstimulation syndrome were treated by ultrasonically guided transvaginal aspiration of ascitic fluid and IV fluid infusion. Ten women with the same condition treated conservatively constituted a comparison group. The main outcome measures included percentage change in hematocrit, creatinine clearance, and urine output before and after aspiration. The duration of hospital stay was compared between the groups. RESULTS: Marked improvement of symptoms and general condition followed soon after aspiration. Hematocrit readings decreased by 22%, creatinine clearance increased by 79.3%, and urine output increased by 220.7%. The average volume of aspirated fluid was 3900 mL. The average duration of hospital stay was 3.8 days in the treated women. In the comparison group, severe symptoms and electrolyte imbalance continued for an average of 9 days, and the average hospital stay was 11 days. CONCLUSION: Intensive IV fluid therapy and transvaginal aspiration of ascitic fluid are safe and effective in improving symptoms, preventing complications, and shortening the hospital stay in severe ovarian hyperstimulation syndrome. PMID- 8416443 TI - Intrauterine device failure: relation to its location within the uterine cavity. AB - OBJECTIVE: To investigate the possible role of position of the intrauterine device (IUD) in accidental pregnancies. METHODS: We examined the location of the IUD in 97 normal women 45-60 days post-insertion, and in 25 pregnant women with the device in situ. RESULTS: A cervically located IUD was identified in seven of 97 women (7.2%) after insertion and in 13 of 25 pregnant women (52%) with the device in situ. The odds ratio for a woman with an intracervical IUD to be pregnant compared with a woman with an IUD in the uterus was 13.93 (95% confidence limits 4.13-48.96). Sonographic follow-up of the pregnant women revealed no change in IUD location during early gestation. CONCLUSIONS: We suggest that cases of failed contraceptive action of the IUD may be secondary to a malpositioned device. A sonographic survey can identify displaced devices. Reinsertion of the IUD in such cases is recommended. PMID- 8416444 TI - Single-dose ampicillin prophylaxis does not eradicate enterococcus from the lower genital tract. AB - OBJECTIVES: To determine the carriage rate of enterococcus in the lower genital tract of women having a cesarean delivery and to determine whether a single 2-g intraoperative dose of ampicillin eradicates enterococcus from the lower genital tract. METHODS: Lower genital tract cultures were taken in 84 women who were in labor or had ruptured membranes and who were about to have an indicated cesarean delivery. The subjects were randomized to receive either a single 2-g dose of ampicillin or a cephalosporin as prophylaxis. Cultures were repeated 24 hours postpartum. RESULTS: Enterococcus was isolated preoperatively in 33 subjects (39.3%) and postoperatively in 36 (42.9%). The enterococcus was eradicated in five of 17 women (29.5%) who received ampicillin. CONCLUSION: These results suggest that a single 2-g dose of ampicillin does not eradicate enterococcus from the lower genital tract. PMID- 8416445 TI - Effects of colchicine on pelvic adhesions associated with the intrauterine inoculation of Neisseria gonorrhoeae in rabbits. AB - OBJECTIVE: To determine whether colchicine, an anti-inflammatory drug, is effective in reducing adhesion formation in a rabbit model in which pelvic inflammation was produced by injection of a suspension of Neisseria gonorrhoeae into the uterine horn. METHODS: Following inoculation, the rabbits were divided into four groups of 11 animals each. The control group received 1 mL saline intramuscularly (IM) for 14 days. Another group received one 100-mg dose of ceftriaxone IM 2-4 hours after inoculation. The third group received one 100-mg dose of ceftriaxone IM 2-4 hours after inoculation and 1 mg colchicine IM daily for 14 days. The fourth group received 1 mg colchicine IM daily for 14 days. The day after the last injection of colchicine, the rabbits were sacrificed and the abdominal cavities were explored to determine the number and grade of adhesions. RESULTS: The incidence of adhesions in the control group was similar to that in the antibiotic-treated group, suggesting that antibiotics do not prevent adhesions in this model. Similarly, the incidence of adhesions in the colchicine and antibiotic group was not different from that in the group receiving colchicine alone. However, the colchicine-treated groups had significantly fewer adhesions than the groups not treated with colchicine (P < .0005). CONCLUSIONS: In this model, antibiotics alone did not prevent the formation of adhesions. Colchicine, alone or in combination with antibiotics, was effective in preventing bacteria-induced pelvic adhesions. Therefore, colchicine may offer a novel approach to the prevention of adhesions associated with pelvic inflammatory disease in women. PMID- 8416446 TI - Comparison of laser-assisted anastomosis, laser welding, and microsurgical anastomosis of rabbit uterine tubes. AB - OBJECTIVE: To assess laser-assisted anastomosis in fallopian tube reconstruction. METHODS: Fifty-two rabbit uterine tubes were transected and subjected to laser welding or to microsurgical or laser-assisted anastomosis in a randomized paired design, and compared with the contralateral side. The time required for the procedure, patency of the uterine tubes, amount of adhesion formation, and quality of scar tissue (grossly and histologically) were evaluated 4 weeks after surgery. RESULTS: Traditional microsurgical procedures required significantly longer operative time than laser-assisted anastomosis (64.4 +/- 2.2 versus 41.3 +/- 2.2 minutes) or laser welding (47.8 +/- 3.0 versus 24.5 +/- 3.0 minutes). Patency was established in all 52 uterine tubes. Adhesion formation was animal dependent rather than procedure-dependent. All scars healed well and most were not visible beyond residue suture. Inflammatory cells were noted around the suture sites; however, gross observation revealed that all mucosal surfaces over the scars were smooth. One of 18 laser-welded uterine tubes developed dehiscence, whereas there was no dehiscence in laser-assisted or microsurgically anastomosed uterine tubes. CONCLUSION: Laser-assisted anastomosis is superior to laser welding or microsurgical anastomosis in rabbit uterine tubes. Laser-assisted anastomosis has potential use in fallopian tube reconstruction. PMID- 8416447 TI - The "less than optimal" cytology: importance in obstetric patients and in a routine gynecologic population. AB - OBJECTIVE: To determine whether patients with less than optimal Papanicolaou tests constitute a low-risk group for developing subsequent abnormalities and thus do not need early repeat screening. METHODS: For the 10-month period October 1989 to August 1990, all screening Papanicolaou tests were classified by the 1988 Bethesda System. Tests designated as less than optimal solely on the basis of lack of an endocervical component were the subject of the study. Prenatal patients with less than optimal tests had repeat tests at the postpartum visit (delayed-repeat group), whereas gynecologic less than optimal tests were repeated within 4 weeks (early-repeat group). The frequency of cytologic abnormalities in our routine gynecologic population was compared with that for both the delayed- and early-repeat testing groups. RESULTS: The less than optimal rate in obstetric patients was 10.2% (153 of 1492), which was significantly higher than the 5.6% rate (473 of 8411) in the routine gynecologic population (P < .0001). The rates of dysplasia or combined abnormalities (dysplasia, human papillomavirus, or atypia) in the delayed-repeat group did not differ significantly from those in the routine gynecologic population (P = .69 and P = .33, respectively). However, the rates of dysplasia or combined abnormalities were significantly lower in the early-repeat group than in the routine gynecologic population (P = .02 and P = .003, respectively). CONCLUSIONS: Less than optimal cervical cytologies occurred almost twice as often in obstetric as in gynecologic patients. Prenatal less than optimal test results were not associated with important cervical pathology, and repeat testing may safely be deferred until postpartum. In addition, early repeat testing in gynecologic patients is a low-yield procedure. PMID- 8416448 TI - Knowledge, attitudes, and practices of obstetricians-gynecologists regarding the prevention of human immunodeficiency virus infection. AB - OBJECTIVE: To determine the knowledge, beliefs, attitudes, and practices of obstetricians-gynecologists regarding human immunodeficiency virus (HIV) prevention. METHODS: Office-based obstetricians-gynecologists in the Washington, DC metropolitan area who reported providing primary care were interviewed by telephone. The survey response rate was 62% (N = 268). RESULTS: The percentages of obstetricians-gynecologists who reported regularly assessing the HIV risk of new adolescent and adult patients were 67 and 40%, respectively. Seventy-two percent reported regularly counseling patients at risk to use condoms for vaginal intercourse, and 60% regularly counseled patients at risk to limit their number of sexual partners. The level of general risk-factor assessment and confidence in the ability to reduce patients' HIV risk were the strongest correlates of the frequency and thoroughness of HIV risk assessment and counseling. CONCLUSIONS: The percentage of obstetricians-gynecologists who assess and counsel patients about HIV risks is below the 75% goal for the year 2000 established by the United States Department of Health and Human Services. Continuing medical education for obstetricians-gynecologists is needed to improve their knowledge and skills in HIV prevention. PMID- 8416449 TI - Laparoscopic sterilization under local or general anesthesia? A randomized study. AB - OBJECTIVE: To assess the safety, acceptability, and economy of local anesthesia and intravenous (IV) sedation versus short-term general anesthesia for laparoscopic sterilization. METHODS: We randomly allocated 125 of 150 consecutively sterilized women to either local or general anesthesia. No women were excluded, but 25 chose not to participate. The women were interviewed before surgery, and they returned a standardized questionnaire after discharge from the hospital. All laparoscopic tubal sterilizations were performed by senior gynecologists. Midazolam was used as premedication. In the local-anesthesia group, lidocaine with adrenaline was infiltrated infraumbilically and bupivacaine was applied to each tube. Midazolam and alfentanil were used as IV sedation. In the general-anesthesia group, intubation anesthesia was accomplished with alfentanil and propofol; atracurium was used for muscle relaxation. RESULTS: In the local-anesthesia group, operation time was shorter, perioperative discomfort was modest, and the costs of equipment were lower than in the general-anesthesia group. There was less postoperative abdominal pain and less need of analgesics, and the patients were more awake in the evening. The rise in heart rate and blood pressure were higher in the local-anesthesia group, and external oxygen was necessary to avoid apnea. Anesthetic surveillance was therefore mandatory. CONCLUSIONS: Local analgesia was highly acceptable to the majority of patients as well as to the gynecologists. The operation time was less, postoperative recovery was quicker, and the women were less bothered by abdominal pain and sore throat. There was a substantial reduction in anesthesia costs. Anesthetic surveillance during surgery was necessary. PMID- 8416450 TI - Adolescent pregnancy in Portugal: effectiveness of continuity of care by an obstetrician. AB - OBJECTIVE: To improve health among newborn infants of pregnant adolescents in Portugal, we developed and evaluated a prenatal care intervention. METHODS: The study group consisted of 80 adolescents recruited as they registered for prenatal care at Hospital Santa Maria in Lisbon. The intervention consisted of three components: initiation of prenatal care at registration, continuity of care by the same obstetrician, and emphasis on the specific nutritional and other health needs of pregnant adolescents. Controls were recruited after delivery on the maternity ward of the same hospital and were matched for age at conception, race, socioeconomic status, years of education, planned versus unplanned pregnancy, and previous body mass index. Controls received the routine care provided to pregnant adult women, including initiation of prenatal care at some point after registration and care by different general practitioners at each visit. RESULTS: Twenty mothers in the control group did not receive prenatal care and were excluded from further analysis. Mothers in the intervention group had their first prenatal visit 2 weeks sooner (P = .02) and had on average almost twice the number of prenatal visits (P = .0001) as controls. They gained 2.0 kg more on average compared with controls (P = .05). Infants of the mothers in the intervention group weighed an average of 181 g more than those in the control group. Fewer infants in the intervention group needed care in the high-risk pediatric unit (P = .005). When potentially confounding variables were controlled in a multivariate analysis, infants in the intervention group weighed on average 174 g more than those in the routine-care group (P = .02). CONCLUSION: This intervention, based on traditional tenets of good medical care, is effective in improving the outcomes of infants born to adolescent mothers in Portugal. PMID- 8416451 TI - Isolated fetal pleural effusion: a prenatal management dilemma. AB - OBJECTIVE: To determine the etiology, pregnancy complications, and outcome of isolated fetal pleural effusion diagnosed antenatally and to evaluate the benefits of prenatal fetal interventions. DATA SOURCES: A literature search of MEDLINE was performed for relevant English language publications between 1985 1991. In addition, reference lists of articles were used to identify reported cases of isolated fetal pleural effusion. METHODS OF STUDY SELECTION: Our search uncovered 31 papers published in peer review journals. From these reports, 82 cases met our selection criteria: All fetuses were diagnosed antenatally with pleural effusion and had no other signs of hydrops at initial diagnosis. DATA EXTRACTION AND SYNTHESIS: The etiology of isolated fetal pleural effusion was unknown in most cases. Possible causes included congenital chylothorax, goiter, lung tumors, and infection. Cardiac defects (4.9%), Down syndrome (4.9%), and polydactyly (1.2%) may be associated with isolated fetal pleural effusion. Perinatal mortality was high (36%) and was related to the development of nonimmune hydrops, prematurity, and pulmonary hypoplasia. Early gestational age at diagnosis of isolated fetal pleural effusion (32 weeks or less) was associated with poor outcome and a neonatal death rate of 55%. In contrast, the neonatal death rate approached 31% as gestational age at diagnosis exceeded 32 weeks. Fifty-four cases were managed conservatively whereas 24 received intrauterine intervention, which included either pleuroamniotic shunt or repeated thoracenteses. Neonatal death rates were 37 and 33%, respectively. CONCLUSION: Not enough data exist to support either the conservative approach or intrauterine pleural drainage in cases of isolated fetal pleural effusion diagnosed antenatally. PMID- 8416452 TI - Laparoscopic trocar-assisted colpotomy. AB - Trocar-assisted transvaginal colpotomy was performed in 17 patients to remove large pathologic specimens during laparoscopic operations when the specimen could not be extirpated through the laparoscopic ports. Under direct laparoscopic visualization, colpotomy was performed using a 10- or 11-mm disposable trocar without the sleeve. The trocar was removed, a ring forceps was placed through the incision, the incision was enlarged, and the mass was grasped with the ring forceps and removed. This technique is safe, fast, and easy to perform. In addition, smoke is not created and potential electrocautery damage to pelvic structures is avoided. PMID- 8416453 TI - Transcervical sonography: an investigational technique for visualization of the embryo. AB - A catheter-based, miniature ultrasound transducer, operating at a frequency of 12.5 MHz, was introduced transcervically into the uterine cavity of 18 pregnant women about to undergo first-trimester abortion. Transcervical sonography showed the yolk sac and embryonic structures, such as brain vesicles, limb buds, liver, spinal canal, and umbilical cord with blood flow, in eight embryos at 5-8 menstrual weeks of age. As an investigational technique, catheter-assisted transcervical sonography offers a new diagnostic approach to imaging of the first trimester human embryo. PMID- 8416454 TI - The relationship of the umbilicus to the aortic bifurcation: implications for laparoscopic technique. PMID- 8416455 TI - Clues to enhancing the identification of human immunodeficiency virus-infected women. PMID- 8416456 TI - Cyst fluid CA 125 levels in ovarian epithelial neoplasms. AB - OBJECTIVE: To assess the feasibility of using cyst fluid CA 125 levels to distinguish between benign and malignant ovarian cystic neoplasms. METHODS: CA 125 levels were measured in ovarian cyst fluid and in serum obtained at surgery in 44 women with ovarian cystic tumors of epithelial origin (25 benign, 12 malignant, and seven borderline). RESULTS: The median cyst fluid CA 125 level in malignant neoplasms (671 U/mL) was higher than in benign tumors (175 U/mL), and 86% of the malignant tumors contained levels higher than 100 U/mL in the cyst fluid, compared with only 62% of the benign tumors. However, these differences were not statistically significant. CONCLUSION: Cyst fluid levels of CA 125 cannot be used to distinguish between benign and malignant ovarian cysts of epithelial origin. PMID- 8416457 TI - The decline of CA 125 level after surgery reflects the size of residual ovarian cancer. AB - OBJECTIVE: To determine whether the decline in CA 125 level after surgery can predict the extent of residual disease. METHODS: In a prospective, nonrandomized clinical trial, 27 women with advanced ovarian cancer (stage III or IV) underwent primary or secondary cytoreductive surgery. CA 125 levels were measured before surgery and 3-14 days postoperatively. The mean postoperative decline was compared by size of residual disease. RESULTS: In women with all visible cancer resected, the mean (+/- standard error of the mean) decline in CA 125 was 91 +/- 4% (1032 +/- 473 U/mL preoperatively to 90 +/- 31 U/mL postoperatively). With less than 2.0 cm residual, there was an 85 +/- 2% decline in CA 125 after surgery (3061 +/- 835 U/mL preoperatively to 456 +/- 146 U/mL postoperatively). In patients with residual disease larger than 2.0 cm, the levels dropped 36 +/- 10% (2958 +/- 1587 to 1904 +/- 708 U/mL). If CA 125 declined less than 60% from the preoperative level, the sensitivity, specificity, and positive and negative predictive values for residual disease larger than 2.0 cm were all 100%. CONCLUSION: By using the ratio of CA 125 levels before and after cytoreductive surgery, one can predict the likelihood that the patient was left with less than 2.0 cm residual disease. PMID- 8416458 TI - Treatment of gonorrhea in pregnancy. AB - OBJECTIVE: To evaluate prospectively the 1989 Centers for Disease Control recommendations for treatment of gonorrhea in pregnancy. METHODS: Two hundred fifty-two women referred with probable endocervical gonorrhea had pre-treatment endocervical, rectal, and oral cultures for Neisseria gonorrhoeae and direct fluorescent antibody testing for Chlamydia trachomatis. They were assigned randomly to receive ceftriaxone 250 mg intramuscularly (IM), spectinomycin 2 g IM, or amoxicillin 3 g orally plus probenecid 1 g orally. Treatment was unblinded and in a 1:1:1 distribution. RESULTS: Two hundred forty-five women (97%) had endocervical infection, 68 (27%) had rectal infection, and 17 (7%) had pharyngeal infection. One hundred two of 252 women (40%) had concomitant endocervical C trachomatis. The overall efficacy was 235 of 252 subjects (93%) (95% confidence interval [CI] 90.1-96.4%). Ceftriaxone was effective in 80 of 84 cases (95%) (95% CI 90.6-99.9%), amoxicillin with probenecid was effective in 75 of 84 cases (89%) (95% CI 82.5-96%), and spectinomycin was effective in 80 of 84 cases (95%) (95% CI 90.6-99.9%). No significant difference was noted in overall efficacy or by site of infection. There was no increased incidence of congenital malformations in the offspring spring of any treatment group. CONCLUSIONS: Ceftriaxone and spectinomycin are safe and effective for the treatment of gonorrhea in pregnancy. Amoxicillin with probenecid has lower efficacy and is not recommended for treatment of gonococcal infection in pregnancy. PMID- 8416459 TI - Birth weight percentiles by gestational age in Canada. AB - OBJECTIVE: To develop current birth weight norms by gestational age for singleton and twin births in Canada. METHODS: Birth weight data were obtained from vital statistics and health department birth registrations for over one million live births in Canada from 1986-1988. Unlikely combinations of birth weight and gestational age were defined within each stratum of multiplicity, gender, and gestational age as records with birth weights more than two interquartile ranges above the 75th percentile or below the 25th percentile. Birth weight percentiles (from first to 99th) by gestational age and sex were calculated for singleton and twin live births. RESULTS: Of the total records, 0.4% were missing data on birth weight or gestational age, and an additional 0.4% were identified as extreme outliers and were excluded from the analysis. Charts of birth weight by gestational age show percentiles 1, 3, 5, 10, 25, 50, 75, 90, 95, 97, and 99 by sex for singleton and twin live births. CONCLUSIONS: The large data base assembled for this analysis provides current, stable birth weight-gestational age percentiles for classifying newborns from a developed country as small, appropriate, or large for gestational age. Compared with birth weight distributions from the 1970s, these current norms are heavier for full-term infants and the interdecile range for preterm infants is narrower. We recommend that birth weight norms be updated every 5-10 years. PMID- 8416460 TI - Maternal hydration increases amniotic fluid index in women with normal amniotic fluid. AB - OBJECTIVE: To test the hypothesis that maternal oral hydration would increase the amniotic fluid (AF) index in pregnancies with normal AF. METHODS: Forty women with a normal AF index (7.0-24.0 cm) were randomized to either the control or hydration group. Women in the hydration group drank 2 L of water and returned for the post-treatment AF index in 4-6 hours, whereas women in the control group drank only 100 mL of water during the same time period. The investigator performing the AF index was blinded to the subject's group. The pre- and post treatment AF indexes and maternal urine specific gravities were compared between the groups. RESULTS: The mean AF index in the hydration group increased significantly by 3.0 +/- 2.4 cm (P < or = .0001) whereas it declined significantly by 1.5 +/- 2.7 cm in the control group (P < or = .02). The maternal urine specific gravities also changed significantly in the expected direction, with those in the hydration group decreasing and those in the control group increasing (P < or = .0001). There was a regression coefficient of -0.6 (P < or = .0001) between the change in urine specific gravity and the change in AF index. The mean time between the pre- and post-treatment AF indexes was not different between the groups. CONCLUSIONS: Maternal oral hydration increased the AF index by approximately 16%, whereas fluid restriction decreased the AF index by 8% in women with normal AF. These findings support previous data that maternal hydration increased the AF index by 31% in women with decreased AF and suggest that maternal fluid volume or osmolality may have a role in maintaining the AF volume. PMID- 8416461 TI - Prenatal diagnosis of gastroschisis: development of objective sonographic criteria for predicting outcome. AB - OBJECTIVE: To determine which sonographic findings predict intestinal damage in fetuses with gastroschisis, and to develop objective criteria that may be useful in selecting candidates for preterm delivery. METHODS: Twenty-four consecutive fetuses at two perinatal centers were assessed retrospectively or prospectively. Maternal, perinatal, and sonographic data were recorded and correlated with postnatal outcome. RESULTS: Bowel diameter of at least 18 mm was associated with a significantly longer time to oral feeding and with significantly greater need for bowel resection. When gestational age was plotted against bowel diameter, a threshold curve was generated, above which all patients had prolonged hypoperistalsis and below which only 30% had prolonged hypoperistalsis. Two infants were delivered at 33 weeks' gestation, both of whom had complications potentially related to prematurity. Only one of 22 patients who delivered later than 33 weeks had similar complications. CONCLUSIONS: Bowel dilatation may be a marker of prenatal bowel damage in fetuses with gastroschisis, especially when it presents late in gestation. Prenatal sonography may be useful in selecting appropriate fetuses for preterm delivery. PMID- 8416462 TI - Ear length in second-trimester aneuploid fetuses. AB - OBJECTIVE: To test the hypothesis that small ears have diagnostic value in detecting second-trimester aneuploid fetuses by ultrasound. METHODS: We prospectively studied 452 patients with singleton pregnancies undergoing ultrasound examination for genetic amniocentesis at 14-25 weeks and an additional 30 singleton pregnancies at 20-25 weeks with a negative anomaly screen. Standard fetal biometry measurements were obtained, including ear length (from helix to end of lobe). RESULTS: Of these patients, 424 (88%) had ear measurements obtained, and a nomogram for ear length by gestational age was compiled. The relationship between ear length and gestational age was linear across the second trimester (r = 0.84, P < .001). Fourteen fetuses had aneuploidy by amniocentesis, of whom ten had ear lengths at or below the tenth percentile. The sensitivity was 71% and the specificity 92% (377 of 410). Positive and negative predictive values were 23% (ten of 43) and 99% (377 of 381), respectively. CONCLUSION: Fetal ear length may be useful in identifying aneuploid fetuses sonographically during the second trimester. PMID- 8416463 TI - Preterm premature rupture of membranes: a randomized study of home versus hospital management. AB - OBJECTIVE: To compare length of latency period, gestational age at delivery, and safety in a carefully selected group of patients with preterm premature rupture of the membranes (PROM) randomized to home versus hospital management. METHODS: After meeting strict inclusion criteria, 67 patients with preterm PROM were randomized by sealed envelope to home versus hospital expectant management. The groups were managed similarly with pelvic and bed rest. Management included recording of temperature and pulse every 6 hours, daily charting of fetal movements, twice-weekly nonstress test and complete blood count, and weekly ultrasound and visual examination of the cervix. RESULTS: There was no significant difference in clinical characteristics or perinatal outcome between the groups. There was, however, a significant decrease in both the days of maternal hospitalization and maternal hospital expenses in the home group. CONCLUSION: Only a very small proportion of cases of preterm PROM (18%) could meet the strict safety criteria for inclusion used in the study. In the home management group, length of the latency period and gestational age at delivery were not significantly different than in hospitalized patients. PMID- 8416464 TI - Preterm delivery among black and white enlisted women in the United States Army. AB - OBJECTIVE: To examine black-white differences in preterm delivery in a healthy population who had unrestricted access to prenatal care. METHODS: We conducted a retrospective cohort study of 842 black and 1026 white enlisted servicewomen who delivered a singleton infant of 20 or more weeks' gestation from July 1, 1987 through September 30, 1990 at four Army Medical Centers in the United States. Data were collected by reviewing maternal and newborn records. We used logistic and proportional hazards regression models to analyze outcomes defined by length of gestation, cause of preterm delivery, and jointly by length and cause. RESULTS: Black enlisted women had a cumulative probability of preterm delivery (13.5%) that was higher than that for white enlisted women (10.5%) (hazard ratio 1.31, 95% confidence interval [CI] 1.002-1.70). However, the ratio of black-to white hazards was not uniform. Black-white differences were small and nonsignificant from 33-36 weeks' gestation, when most preterm deliveries occur. The differences were also small and nonsignificant for deliveries related to spontaneous rupture of membranes or idiopathic preterm labor, the most common causes of preterm delivery. The black-to-white hazard ratios were greatest for all deliveries before 33 weeks' gestation and for medically indicated preterm deliveries. CONCLUSIONS: Efforts to reduce black-white differences in preterm delivery must go beyond providing prenatal care and eliminating recreational drug use. Future studies should consider black-white differences in environments during the mother's own development and in psychosocial and physical stresses during pregnancy. PMID- 8416465 TI - The effect of prophylactic heparin treatment on enhanced thrombin generation in pregnancy. AB - OBJECTIVE: To assess the activation status of blood coagulation in thrombosis prone pregnant women receiving a reduced dosage of heparin. METHODS: Forty-three women were given subcutaneous heparin treatment during pregnancy; the dosage was monitored so that the heparin (anti-Xa) level was within the stipulated range of 0.08-0.15 anti-Xa units 3 hours after injection. Several coagulation variables were investigated and some routine analyses were performed. The results were compared with those of a control group of 26 healthy pregnant women. RESULTS: Compared with the control values, hemoglobin (P < .01), fibrinogen (P < .05), and total protein S (P < .01) were increased already before heparin treatment was started and continued to be increased significantly throughout pregnancy. During heparin treatment, protein C levels (P < .01) and the factors measured by prothrombin time (P < .001) were increased compared with the controls. Antithrombin decreased, though not as much as when patients are given therapeutic dosages. The fibrinolytic inhibitors behaved as in normal pregnancy. Three variables measuring thrombin formation or coagulation activation (ie, thrombin antithrombin complexes, D-dimers, and soluble fibrin) were increased in a high proportion before heparin treatment was started; they normalized somewhat during treatment. CONCLUSIONS: In early pregnancy, women with previous thrombotic episodes have high plasma levels of coagulation variables and of markers of coagulation activation compared with controls. Such changes may be used to predict the need for treatment and, in the future, to control treatment. PMID- 8416466 TI - Measurement of embryotoxic factors is predictive of pregnancy outcome in women with a history of recurrent abortion. AB - OBJECTIVE: To determine whether the production of embryotoxic factors in early pregnancy is predictive of pregnancy outcome in women with a history of two or more unexplained recurrent abortions. METHODS: Between July 1987 and June 1991, 450 nonpregnant women were evaluated for recurrent abortion. Embryotoxic factors were found in 328 of 346 women with an otherwise unexplained etiology. These women were given progesterone immunosuppressive therapy in a subsequent conception cycle, and embryotoxic factors were measured at 5 weeks' gestation. RESULTS: Data on embryotoxic factors and pregnancy outcome were available for 141 of 208 reported subsequent pregnancies. Of women still positive for embryotoxic factors, 40 had a repeat first-trimester spontaneous abortion whereas 16 delivered a viable infant; of women who no longer produced embryotoxic factors, only 11 had a repeat first-trimester abortion and 74 delivered a viable infant. CONCLUSIONS: The embryotoxic factor assay may be useful in predicting pregnancy outcome in women with a history of unexplained recurrent abortion. Further work is needed to evaluate the effectiveness of immunosuppression for this condition. PMID- 8416467 TI - Glycosaminoglycans in cervical connective tissue during pregnancy and parturition. AB - OBJECTIVE: To investigate the content and distribution patterns of glycosaminoglycans in the human cervix during pregnancy and parturition. METHODS: We obtained a total of 87 specimens from nonpregnant and pregnant women. Biopsies (weight 50-200 mg) were taken from the posterior lip of the cervix. Hyaluronic acid, dermatan sulfate, chondroitin sulfates, and heparan sulfate were separated on a Dowex 1 x 2 column and identified. RESULTS: The total amount of glycosaminoglycans increased during pregnancy from 2800 to 5000 nmol/g dry weight. The highest values were observed at the onset of labor (7100 nmol/g dry weight), followed by a sharp decrease during parturition. The clinical features of cervical ripening and dilatation were also associated with remarkable changes in glycosaminoglycan patterns. CONCLUSION: Besides collagenolysis during pregnancy, the glycosaminoglycans are also important regulators of cervical function. The different clinical features of the human cervix are characterized not only by variation in the total glycosaminoglycan content but also by changes in the proportions of the different glycosaminoglycans. PMID- 8416468 TI - Essential fatty acids in the treatment of premenstrual syndrome. AB - OBJECTIVE: To determine whether essential fatty acids are effective in the treatment of premenstrual syndrome (PMS). METHODS: In a randomized, double-blind, crossover trial, we studied 27 women diagnosed with PMS over ten menstrual cycles and 22 symptom-free controls over one cycle. The first cycle was used for diagnostic assessment. For the women with PMS, placebos were administered during the second cycle. This was followed by randomization to four cycles of active treatment with essential fatty acids and four cycles of placebo, with a crossover after completion of the fourth cycle. Assessment of symptoms and diagnosis of PMS were based on daily self-ratings made by the women throughout the study. RESULTS: Treatment with essential fatty acids did not reduce premenstrual symptoms or symptom cyclicity. However, time had a significant effect on a number of symptoms, indicating either a placebo effect or an effect from participation in the study. Women with PMS had a significantly higher frequency of dysmenorrhea and familial PMS than did the symptom-free controls. CONCLUSION: Treatment with essential fatty acids is ineffective therapy for PMS. The improvement we observed over time can be ascribed to either a placebo effect or participation in the study. PMID- 8416469 TI - CA 125 levels measured in different phases of the menstrual cycle in screening for endometriosis. AB - OBJECTIVE: To examine variations in CA 125 levels during the three phases of the menstrual cycle in women with and without endometriosis. METHODS: One hundred infertile women were studied prospectively. CA 125 levels were measured during menses and during the follicular and luteal phases before diagnostic laparoscopy. Subjects were divided into four groups: no evidence of endometriosis (35 women), stage I endometriosis (30 women), stage II endometriosis (21 women), and stages III and IV endometriosis (14 women). RESULTS: In the endometriosis groups, there was a significant difference in the mean CA 125 levels drawn at menses and those drawn in the follicular phase. In patients with severe endometriosis, there was also a difference in the mean CA 125 levels drawn at menses and in the luteal phase. This finding led to the development of a screening test based on the ratio of CA 125 levels at menses to levels in the follicular phase. The test based on this ratio (with a cutoff of 1.5) had a sensitivity of 62.5% and specificity of 75%, compared with a sensitivity of 26.8% and specificity of 100% for the test based on a single CA 125 level drawn at menses (with a cutoff of 35 U/mL). CONCLUSIONS: CA 125 levels during menses are elevated compared with those during the follicular phase in patients with endometriosis. Screening tests based on the relationship of multiple CA 125 levels taken throughout the menstrual cycle were more sensitive for detection of endometriosis than tests based on a single CA 125 level. PMID- 8416470 TI - Missed opportunities for childhood vaccinations in office practices and the effect on vaccination status. AB - To determine the rate of childhood under-vaccination, rate and types of missed opportunities (MOs) for vaccinations, and the contribution of MOs to the undervaccination of preschool-age children, the authors conducted a retrospective medical chart review in seven primary care settings in the Rochester, NY, area: a hospital clinic, a neighborhood health center, a group-model health maintenance organization, an urban group practice, a suburban group practice, a rural health center, and a rural private practice. The random sample included 1124 children having birth dates between March 15, 1988, and September 15, 1989. The main outcome measures were cumulative undervaccination rate, defined as the proportion of patients from each practice who were ever > 60 days past-due for a vaccination by 12, 18, or 24 months of age; undervaccination time, defined as the median number of months during which children were undervaccinated; number of MOs; visit types and conditions associated with the MOs; and the duration of undervaccination time attributable to MOs. The cumulative undervaccination rate by 12 months was at least 20% in each practice except for the suburban practice, where it was 4%. The frequency of MOs varied from a high of 1.8 MO per patient per year at the rural private practice to a low of 0.3 MO per patient per year at the suburban practice. More than one quarter of MOs occurred during either health supervision or follow-up visits in all practices. In 28% of visits during which an MO occurred, patients had no fever or acute illness.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416471 TI - Effects of antiepileptic drugs on reaction time, attention, and impulsivity in children. AB - Simple, choice, and complex reaction times, attention (variability of responses and omission errors), and impulsivity (commission and wrong-hand errors on choice and complex reaction time) were repeatedly measured in 111 epileptic children, aged 5 to 13 years, tested a total of 232 times. Antiepileptic drugs (AEDs) were started, stopped, and adjusted throughout the study period, for a variety of clinical indications, and AED serum levels were monitored. The relationship of performance to AED serum level was examined. Overall the nonspecific effect of AEDs was minimal: higher total serum levels of AEDs correlated with more impulsive errors on complex reaction time testing only. In contrast, in 54 children receiving carbamazepine monotherapy, we found a dose-related beneficial effect upon reaction time, with higher serum levels associated with faster responses and fewer omission errors, particularly on complex reaction time. Phenobarbital caused minimal dose-related effects: only variability and impulsive errors increased with increasing serum levels, and only on one segment of the test (73 subjects). PMID- 8416472 TI - Relationship between epidemiologic risk factors and clinicopathologic findings in the sudden infant death syndrome. AB - The risk of sudden infant death syndrome (SIDS) is said to be enhanced by factors such as prematurity, low birth weight, and perinatal distress. The significance of risk factors for SIDS research was questioned because the majority of SIDS victims seem to lack them. Therefore, postmortem records of 1144 infants who died suddenly and unexpectedly in King County, Washington, over a 25-year period were studied. Deaths were classified as "explained" if a cause was apparent, "classic" SIDS if the history and autopsy were unrevealing or, where the diagnosis of SIDS was doubtful, as "probable" or "possible" SIDS. The infants' birth certificates were compared with those of 3647 infants born during a similar period. Seventy nine deaths (7%) were explained. The 1065 previously certified as SIDS were reclassified classic SIDS (82%), probable SIDS (13%), and possible SIDS (5%). Low birth weight, small size for gestational age, prematurity, and low 5-minute Apgar scores each form a "continuum"; the possible-SIDS group had the highest proportion of such infants, followed by the probable- and classic-SIDS groups, which exhibit extensive overlap with the control population. A 5-minute Apgar score of less than 7 and delayed postnatal growth rate are not risk factors for classic SIDS. Risk factors are more prevalent in SIDS infants where the diagnosis may be doubtful. The great majority of SIDS victims possess fewer risk factors. To avoid the bias of confounding variables, SIDS research should focus on as "pure" a SIDS population as is possible. PMID- 8416473 TI - Predicting preschool behavior problems from temperament and other variables in infancy. AB - There is uncertainty about the relationship between difficult temperament in infancy and reported problem behaviors later in childhood. In this study data from a large, representative community cohort (total N studied = 1583) were used to determine whether preschool behavior problems (at age 4 to 5 years) could be predicted from difficult temperament and other variables in infancy. Maternal ratings of difficult temperament on the Revised Infant Temperament Questionnaire predicted only 17.5% of those with preschool behavior problems, a percentage not significantly greater than the 14% of the total sample rated as having problems. There was some improvement in prediction when difficult temperament was added to other variables such as male sex (28%). However, mothers' overall rating of temperament was a more powerful predictor of preschool behavior problems, both alone (26.0%) and in combination with other variables such as perinatal stress (36.8%), male sex (29.5%), and non-Australian parent (29.4%). Similarly, maternal reports of infant behavior problems was a more powerful predictor of preschool behavior problems both alone (21.8%) and in combination with male sex (24.6%), low socioeconomic status (26.1%), non-Australian parent (21.8%), and nurse's overall rating of temperament (21.8%). The best consistent predictor of later problems was the combination of mothers' overall rating of temperament and maternal reports of infant behavior problems (27.0%), especially when combined with other infant variables such as perinatal stress (35.3%), male sex (31.5%), and non-Australian parent (30.0%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416474 TI - Foreign body ingestions in children: risk of complication varies with site of initial health care contact. Pediatric Practice Research Group. AB - Current recommendations for the management of pediatric foreign body ingestions are based on studies of patients cared for at tertiary care hospitals; they call for aggressive evaluation because of a high incidence of complications. Two hundred forty-four children with suspected foreign body ingestions were prospectively followed to analyze adverse outcomes, ie, procedures, complications, and hospitalizations. Patient enrollment into the study was from three sources: (1) patients who referred themselves to a tertiary pediatric emergency department, (2) patients referred to the same tertiary pediatric emergency department after an initial evaluation by another hospital or physician, and (3) patients who reported their foreign body ingestions to a private pediatric practitioner participating in the study. Most children were well toddlers in normal circumstances, under parent supervision at the time of ingestion. Coins were the most common item ingested (46%). Procedures were done in 53 (24%) of 221 patients and complications occurred in 48 (22%) of 221. Complications were higher in patients referred to the emergency department (63%) than in emergency department self-referred patients (13%) or private practice patients (7%) (chi 2, P < .01). These findings demonstrate the risk of drawing conclusions regarding a universal standard of care from studies involving only hospital-based patients. PMID- 8416475 TI - Electrographic seizures in preterm and full-term neonates: clinical correlates, associated brain lesions, and risk for neurologic sequelae. AB - Electrographically confirmed seizures in preterm and term neonates were compared with respect to clinical correlates, incidence, associated brain lesions, and risk for neurologic sequelae. Over a 4-year period, 92 neonates from a neonatal intensive care unit population of 4020 admissions at a large obstetric hospital with 40,845 livebirths had electrographically confirmed seizures. Sixty-two neonates were preterm and 30 were full-term for gestational age. Chi-square calculations were used to compare the two groups. While the incidence of seizures for all neonates admitted to a neonatal intensive care unit was 2.3%, outborn neonates were more likely to have seizures than inborn neonates. Preterm neonates of < or = 30 weeks gestational age had a seizure frequency of 3.9%, which was significantly higher than that of older preterm neonates and full-term neonates. Clinical criteria contemporaneous with electrographic seizures were noted in only 28 (45%) of 62 preterm, and 16 (53%) of 30 full-term neonates. Subtle seizures coincident with electrographically confirmed seizures were the most predominant clinical type for both term and preterm neonates (71% and 68%, respectively). The distribution of clonic, myoclonic, and tonic seizures was also similar for both groups. Autonomic signs coincident with electrographically confirmed seizures (ie, blood pressure, heart rate, oxygenation, respiration changes) were more frequently observed in preterm than full-term neonates with subtle seizures; 7 (37%) of 19 compared with 1 (6%) of 16. Electrical seizures without clinical correlates were noted more frequently than electroclinical seizures for both populations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416476 TI - Bronchial hyperresponsiveness in children with atopic dermatitis. AB - Using histamine provocation tests, bronchial responsiveness was measured in 43 children with atopic dermatitis, aged 7 to 15 years, who had attended pediatric dermatology clinics within the past 5 years. The children were divided into two groups, one group of 21 children in whom asthma had been previously diagnosed and in whom symptoms of asthma had occurred in the preceding year, and the other comprising 22 children who denied any such symptoms. Bronchial hyperresponsiveness (as defined by a 20% fall in forced expiratory volume in 1 second at a provoking dose of histamine of 7.8 mumol or less [PD20]) was demonstrated in all but 1 of the children with atopic dermatitis and asthma, in 18 of the 22 children with atopic dermatitis alone, but in only 3 of a control group of 18 children without atopic dermatitis or asthma. The asthmatic subjects (median PD20 = 0.22 mumol) had more severe bronchial hyperresponsiveness than the nonasthmatic subjects (median PD20 = 2.10 mumol). This study confirms the strong association between atopic dermatitis and bronchial hyperresponsiveness, even in the absence of overt asthmatic symptoms. PMID- 8416477 TI - Inadvertent administration of positive end-distending pressure during nasal cannula flow. AB - In the clinical setting, nasal cannulas are frequently used to deliver supplemental oxygen to neonates and are not believed to affect the general respiratory status. In contrast, it was hypothesized that clinical changes associated with nasal cannula gas flow may be related in part to the generation of positive end-distending pressure. To test this hypothesis, alterations in esophageal pressure were quantified as an indication of end-distending pressure and thoracoabdominal motion was quantified as an indication of breathing patterns in 13 preterm infants at gas flow levels of 0.5, 1, and 2 L/min delivered by nasal cannula with an outer diameter of either 0.2 or 0.3 cm. Changes in esophageal pressure were assessed by esophageal balloon manometry. Ventilatory patterns were assessed from thoracoabdominal motion by using respiratory inductive plethysmography. Thoracoabdominal motion was quantitated as a phase angle (theta); larger values represent greater asynchrony. The 0.2-cm nasal cannula did not deliver pressure or alter thoracoabdominal motion at any flow. In contrast, the 0.3-cm nasal cannula delivered positive end-distending pressure as a function of increasing levels of gas flow (r = .92) and reduced thoracoabdominal motion asynchrony. The mean pressure generated at 2 L/min was 9.8 cm H2O. These data demonstrate that nasal cannula gas flow can deliver positive end-distending pressure to infants and significantly alter their breathing strategy. This finding raises important concerns about the indiscriminate therapeutic use, size selection, and safety of nasal cannulas for the routine delivery of oxygen in preterm infants. PMID- 8416478 TI - Treatment of attachment disorder of infancy in a neonatal intensive care unit. PMID- 8416479 TI - Dishwasher effluent burns in infants. PMID- 8416480 TI - Neonatal hyperparathyroidism and skeletal demineralization in an infant with familial hypocalciuric hypercalcemia. PMID- 8416481 TI - The senior pediatrician and day care. PMID- 8416482 TI - Home phototherapy: not seeing the light. PMID- 8416483 TI - Group B streptococcal infections: an obstetrical viewpoint. PMID- 8416484 TI - Thrombocytopenia associated with acute hepatitis B infection. PMID- 8416485 TI - Necrotizing lymphadenitis (Kikuchi's disease). PMID- 8416486 TI - Retraction of the umbilicus during voiding as an initial sign of a urachal anomaly. PMID- 8416487 TI - Sample size: profound implications of mundane calculations. PMID- 8416488 TI - American Academy of Pediatrics Committee on School Health. Basic life support training school. PMID- 8416489 TI - Could the Childhood Vaccine Act be bad? PMID- 8416490 TI - Mythology of lead poisoning. PMID- 8416491 TI - Mythology of lead poisoning. PMID- 8416492 TI - SSI benefits and functional assessment. PMID- 8416493 TI - Chronic fatigue in children: illness or disease? PMID- 8416494 TI - Questions about bilirubin. PMID- 8416495 TI - Aural, oral, or rectal--does it make any real difference? PMID- 8416496 TI - New recommendations not very different from current practice. PMID- 8416497 TI - Overtreatment of neonates. PMID- 8416498 TI - Overtreatment of neonates. PMID- 8416499 TI - Modified chickenpox in children immunized with the Oka/Merck varicella vaccine. AB - Oka/Merck varicella vaccine has been studied in this institution since 1981. Persistence of antibody for 6 to 8 years has been demonstrated; however, cases of chickenpox have been seen in immunized children. The severity of chickenpox in healthy children who have received Oka/Merck varicella vaccine since 1981 is described. All vaccinees who developed chickenpox-like rashes more than 6 weeks postimmunization were examined. Of 2163 vaccinees, 164 were examined, of whom 114 had rashes consistent with chickenpox. When sera were available (46%), antibody studies uniformly confirmed varicella-zoster virus infection. Chickenpox occurred 2 to 96 months (median of 44 months) postimmunization. The range for the number of skin lesions was 1 to 285 (median 18) in seroconverters. Symptoms included itching in 39%, fever in 9%, headaches in 7%, lymphadenopathy in 3%, and malaise in 2%; 54% were asymptomatic, except for the rash. The median time to total healing was 5 days. The median time lost from school was 2 days. Thirteen of the children in whom infections developed had failed to seroconvert after immunization. Their infections were similar in severity to those of children who had seroconverted originally. When varicella was introduced into families as a result of chickenpox in an immunized family member (index case), the rate of secondary chickenpox among immunized siblings was 12.2%. Eleven such secondary cases were similar in severity to the 9 index cases. It is concluded that chickenpox is generally mild in previously immunized children. PMID- 8416500 TI - Overtreatment of neonates. PMID- 8416501 TI - Overtreatment of neonates. PMID- 8416502 TI - Single-dose intramuscular ceftriaxone for acute otitis media in children. AB - This study evaluated the efficacy of a single dose of intramuscular ceftriaxone for acute otitis media in children, using amoxicillin as a control. (There is currently no established single-dose treatment for this condition.) In a prospective, randomized, double-blind, clinical trial, 233 children, aged 5 months to 5 years, with uncomplicated acute otitis media were randomly assigned to receive either a single intramuscular injection of ceftriaxone (50 mg/kg) plus placebo oral suspension for 10 days, or a placebo injection plus amoxicillin oral suspension (40 mg/kg per day divided three times per day) for 10 days in a double blind fashion. Demographic and clinical characteristics were similar in both groups. Treatment was successful in 107 of 117 given amoxicillin (91%, 95% confidence interval 86% to 97%) and 105 of 116 given ceftriaxone (91%, 95% confidence interval 85% to 96%). Rates of improvement, failure, relapse, and reinfection were similar in both groups, as were the otoscopic and tympanometric evaluations at the 14- and 60-day follow-up visits. It is concluded that a single intramuscular injection of ceftriaxone (50 mg/kg) is as effective as 10 days of oral amoxicillin for the treatment of uncomplicated acute otitis media in children. PMID- 8416503 TI - Transmission of human genital papillomavirus disease: comparison of data from adults and children. AB - A substantial body of evidence has demonstrated that the primary means of transmission of genital warts in sexually active adults is through sexual contact. However, the epidemiology and social significance of anal-genital warts in prepubertal children is controversial. Debate continues regarding the frequency with which these lesions have resulted from sexual abuse or transmission by other means. An accurate understanding of the dominant means of transmission of anal-genital warts in children is of particular importance because that understanding influences the extent to which child protective services may become involved following a diagnosis. This paper reviews the evolution of the data on the means of transmission of human papilloma virus disease of the genital tract of adults and compares those data with the information available concerning the transmission of anal-genital human papillomavirus-related disease in children. Methods for the diagnosis of child sexual abuse that have developed in the past decade form one of the bases for the evaluation of studies of the transmission of anal-genital human papillomavirus related diseases to children. PMID- 8416504 TI - Situational and sociodemographic characteristics of children infected with human immunodeficiency virus from pediatric sexual abuse. AB - This study assessed the situational and sociodemographic characteristics of children infected with human immunodeficiency virus (HIV) from pediatric sexual abuse. A letter of inquiry was sent to 2147 professionals across health and social service disciplines involved with child abuse assessment, treatment, and prevention. Respondents working in programs where HIV antibody testing of abuse victims occurs and who had identified HIV infection in one or more abused children were sent a survey to assess the demographics of victims, the family/living situation where abuse occurred, alternative risks for HIV infection, bases for diagnosis of sexual abuse and for HIV antibody testing, and profiles of the perpetrator and type of abuse. Of 5622 estimated HIV antibody tests conducted during 113,198 sex abuse assessments, 28 children were infected with HIV and lacked any alternative transmission route to that of sexual abuse. A total of 41 HIV-infected children with a history of sexual abuse were identified. Thirteen cases had alternative risk factors and were excluded from analysis. Sixty-four percent of the 28 victims with sexual abuse as the sole risk factor were female and 71% were African-American. The mean age was 9 years. Coinfection with another sexually transmitted disease (STD) occurred in 9 (33%) cases. Sexual abuse was diagnosed on the basis of a victim disclosure in 21 (75%) cases. The basis for HIV antibody testing was physical findings suggestive of HIV infection in 9 (32%) cases, HIV-seropositive or high-risk perpetrator in 6 (21%) and 2 (7%) cases, respectively, and the presence of another STD in the victim in 4 (14%) cases. Perpetrators were a child's parent in 10 (42%) cases and another relative in 6 (25%) cases.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416505 TI - Childhood asthma and poverty: differential impacts and utilization of health services. AB - Data from the 1988 National Health Interview Survey on Child Health showed that 4.3% of all children younger than 17 years of age had asthma, with poor children demonstrating a slightly greater prevalence than nonpoor (4.8 poor vs 4.2 nonpoor). This poor/nonpoor differential was even greater for children younger than 6 years old (4.2 vs 3.1). Poor children were also more likely to have had more than 7 bed days in the past year because of their asthma. Measures of health service utilization showed that poor children had 40% fewer doctor visits (3.2 vs 5.4) and had 40% more hospitalizations in the previous year (10.6% vs 7.4%). Although more than 90% of all children had a usual source of routine and/or sick care, poor children were more likely to receive routine care in a neighborhood health center (15.1% vs 1.6%) or hospital-based clinic (11.1% vs 2.8%) than in a doctor's office (67.2% vs 91.1%) and, when sick, then were more than four times more likely to report an emergency department as a usual source of care (8.1% vs 1.7%). Diminished accessibility to appropriate outpatient health services for poor children with asthma was associated with increased morbidity, measured by hospitalization, and bed days. These findings have significant implications for the development of comprehensive models of care and the potential role that community clinics could play with increased funding as a result of Medicaid changes that were instituted as part of the 1989 Omnibus Budget Reconciliation Act. PMID- 8416506 TI - Hispanic children with asthma: morbidity. AB - Hispanic children represent a large and growing segment of the poor and disadvantaged children in our country. Asthma and other chronic respiratory diseases have a significant impact on poor children. Yet there are few descriptions of the specific morbidities and barriers to health that Hispanic children with asthma encounter, and data on predictors of morbidity among these children are unavailable. The purpose of this study is to describe the morbidity associated with asthma in Hispanic children and to identify factors that predict morbidity. A group of Hispanic children with moderate asthma followed in the clinics of the University of Texas Health Science Center at San Antonio were studied. Children aged 6 to 16 years with at least two acute-care visits or one hospitalization for asthma during the previous year were enrolled. Data sources included standardized questionnaires, spirometry, medical records, and school attendance records. Seventy-eight Hispanic children were enrolled in the study (mean age = 9.4 +/- 2.7 [SD]; 62% male). Fifty-two (67%) of children had been hospitalized previously. The other morbidity variables (mean +/- SD) were number of days/week impaired (1.1 +/- 1.2), number of days absent from school per year (13 +/- 9.6), number of acute-care visits per year (3.3 +/- 2.4), and number of hospital admissions per year (0.6 +/- 0.8). The mean forced expiratory volume in 1 second/forced vital capacity was 79.3% (+/- 9.1) and the mean forced expiratory flow, mid-expiratory phase, percent predicted was 69.9% (+/- 25.1). Thirty-four children (44%) were exposed to cigarette smoke in the home.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416507 TI - Appropriateness of hospitalization in a Canadian pediatric hospital. AB - Allocation of limited resources in the Canadian health care system is hampered by a lack of studies addressing the appropriateness of the pediatric patient days in hospital. The authors retrospectively reviewed one hospital day per month in 1988, using a Pediatric Appropriateness Evaluation Protocol previously used in the United States. Of 878 inpatients, 852 charts were reviewed, and 26 charts were unavailable for study. The patients ranged in age from premature newborns to 20 years old. There were 475 medical days, 359 surgical days, and 18 patients to other services. Statistical significance was tested using the chi 2 test for contingency tables. Twenty-four percent of patient days were inappropriate. Younger children and shorter lengths of stay were more likely to result in appropriate hospital days. For infants younger than 60 days, 11% of days in hospital were inappropriate, 21% of days for infants between 2 months and 1 year of age, 25% for children between 1 and 5 years, and 36% for children older than 5 years of age. Children hospitalized 2 days or less had inappropriate hospital days accounting for 16% of the reviewed days. This increased to 33% for 3 to 14 days of hospitalization. Inappropriate hospital days did not vary significantly from month to month. Surgical patients had more appropriate hospital days than medical patients. Admission route (elective, emergency, or transferred from another hospital) did not affect the appropriateness of the subsequent day reviewed. It is concluded that inappropriate hospitalization in a Canadian pediatric hospital occurs only slightly more frequently than in an American pediatric hospital.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416508 TI - Patterns of smokeless tobacco use by young adolescents. AB - The use of smokeless tobacco products by adolescents has reportedly increased. The purpose of this study was to examine the use of smokeless tobacco by young adolescents across geographic locations and to look at patterns of use and variables associated with continued use beyond experimentation. Participants were 2018 students in sixth through ninth grades. Forty-five percent were male and 76% were white. Use of smokeless tobacco products was reported by 12% of the total population, and 25% reported smoking. Smokeless tobacco use was associated with cigarette smoking, alcohol use, and parental substance abuse. Those reporting alcohol use were more than four times more likely to be users of smokeless tobacco than nondrinkers. We found increased age, being male, being white, smoking, drinking, perceived effects of smokeless tobacco use, and friends' smoking behavior to be significantly associated with continued use beyond experimentation. Smokeless tobacco use was reportedly greater in rural areas. Adolescents who reported initiating use between 6 and 8 years of age were using on a more frequent basis than those who had initiated use when older. This study demonstrated the need for targeting elementary schools for educational interventions aimed at reducing smokeless tobacco use. PMID- 8416509 TI - Prevalence of respiratory symptoms in children with atopic dermatitis attending pediatric dermatology clinics. AB - A postal questionnaire, completed by parents, was used to study the prevalence of respiratory symptoms in 250 children with atopic dermatitis who had attended pediatric dermatology clinics in London within the past 5 years. A response rate of 84% was achieved. Each child had a control matched for age and sex, whose parents filled in an identical questionnaire. The prevalence of wheezing was 76% in the atopic dermatitis group and 12% in the control group. Of the children with atopic dermatitis who wheezed, 87% had been given a diagnosis of asthma by a doctor, compared with 40% of the control children who wheezed. Overall, 68% of the children with atopic dermatitis had been given a diagnosis of asthma by a doctor. This prevalence of respiratory symptoms and of diagnosed asthma is substantially higher than has been shown in previous studies. As the test population consisted of children who had been referred to a tertiary center for management of their skin disease, this higher prevalence may partially reflect the increased severity of atopic dermatitis in the study group, as well as the heightened awareness of the association between these two diseases by their parents and physicians. PMID- 8416510 TI - Toxic hydroperoxides in intravenous lipid emulsions used in preterm infants. AB - The unsaturated fatty acids that make up a large component of the lipid emulsion Intralipid are highly susceptible to peroxidation, and the products of this reaction could explain the toxicity that has been associated with the administration of some emulsions. Lipid peroxidation produces hydroperoxides, which can alter arachidonic acid metabolism or react to form organic free radicals, which then stimulate a cascade of damage to endogenous lipids. The lipid hydroperoxides and their breakdown products are also mutagens and carcinogens. To determine the degree of lipid peroxidation in Intralipid, we measured the lipid hydroperoxide content of three lots of 20% Intralipid using high-performance liquid chromatography with chemiluminescence detection. The average concentration was 290 +/- 29 mumol/L (SEM) lipid hydroperoxides (n = 15), a large portion of which was made up of trilinoleate derivatives. Measurements made on Intralipid samples collected from the end of the intravenous tubing after a 20-hour infusion cycle were not significantly different from measurements made on newly opened bottles. The lipid hydroperoxide content of some lipid emulsions may represent a clinically significant risk to premature infants, particularly those with preexisting lung disease. PMID- 8416511 TI - Comparison of maternal sera, cord blood, and neonatal sera for detecting presumptive congenital syphilis: relationship with maternal treatment. AB - The incidence of congenital syphilis has increased rapidly over the past few years. Most infected mothers and their newborns are asymptomatic at birth and diagnosis depends on serologic testing during pregnancy and at delivery. This study was initiated to compare maternal sera, cord blood, and neonatal sera for detecting presumptive congenital syphilis and to assess the role of maternal treatment (administration of penicillin to the mother at least 1 month before delivery) on the serologic results at the time of delivery. The serologic results from all live deliveries complicated by a positive maternal and/or neonatal test for syphilis during a 12-month period were compared using chi 2 analysis and multiple comparisons for proportions. Of 3306 livebirths, 73 (2.2%) were complicated by a positive maternal or neonatal serology. At delivery, the serologic test was positive in 68 (94%) of 72 maternal sera, 30 (50%) of 60 cord sera, and 43 (63%) of 68 neonatal sera. In the absence of maternal treatment, 95% of the maternal sera, 66% of the cord blood samples, and 86% of the neonatal sera were positive. If the mother had been treated, 94% of maternal sera, 36% of cord sera, and 39% of neonatal sera were positive. Cord blood and neonatal sera appear to be inferior to maternal sera for detecting prenatal exposure to syphilis. Cord serology is also inferior to neonatal serology at 2 to 3 days of age. The most effective way to identify newborns at risk for congenital syphilis is to obtain a maternal serologic diagnosis during pregnancy and to test maternal and neonatal sera at delivery. PMID- 8416512 TI - Primary hyperlipidemia in a pediatric population: classification and effect of dietary treatment. AB - Patients seen at a pediatric lipid clinic over a 27-month period were reviewed retrospectively to evaluate types of primary lipid disorders and effect of dietary treatment at the first follow-up visit. One hundred eighty-two patients were classified into one of four categories: (1) elevated low-density lipoprotein cholesterol (LDL-C) with LDL-C > 95th percentile (32%); (2) isolated triglyceride (TG)/high-density lipoprotein cholesterol (HDL-C) abnormalities, with TG > 95th percentile and/or HDL-C < 5th percentile (30%); (3) borderline LDL-C, TG, or HDL C (29%); (4) normal (9%). The American Heart Association Step-One Diet was prescribed for all patients older than 2 years, and they received extensive nutritional and risk-management counseling. Of these patients, 59 (32%) returned for at least one follow-up visit and mean changes in lipid values between initial and first follow-up visits were evaluated. Levels of LDL-C decreased by 24 mg/dL in 22 patients with elevated LDL-C levels. Triglyceride levels decreased by a mean of 22 mg/dL and HDL-C increased by a mean of 4 mg/dL in 21 patients with isolated TG/HDL-C abnormalities. Levels of LDL-C tended to rise in this group, but not to a significant degree. A new finding of this report is that screening for total cholesterol results in the identification of many children with TG or HDL-C abnormalities alone and that the Step-One Diet appears to be effective in improving both TG and HDL-C levels in these patients. PMID- 8416513 TI - Home phototherapy: use and attitudes among community pediatricians. AB - Rather than using home phototherapy (HP), many pediatricians admit neonates to the hospital for the treatment of unconjugated hyperbilirubinemia. This study investigates the concerns and experiences of pediatricians related to HP use. A questionnaire was sent to 150 pediatricians in the Philadelphia, PA, area and 94 returned questionnaires. Of the responders, 30 of 94 had used HP. Twenty-nine users and 62 of 64 nonusers answered detailed questions regarding HP. Few practitioners follow American Academy of Pediatrics guidelines in selection of HP candidates. Among 29 pediatricians, prematurity (7), Rh incompatibility (13), positive direct Coombs (21), and ABO incompatibility (22) were not considered contraindications, although the American Academy of Pediatrics recommends all as contraindications. Almost all (29/30) HP users identified at least one problem with home treatment. The most commonly reported problems were parental anxiety, blood testing difficulties, and reimbursement issues. Nonusers (62) were asked their reasons for not using HP. The most common replies related to concerns about noncompliance (25), medical-legal issues (23), and administrative difficulties (22), none of which were cited as problems by HP users. PMID- 8416514 TI - The cloverleaf skull anomaly: managing extreme cranio-orbitofaciostenosis. AB - The cloverleaf skull anomaly represents the most manifestly extreme form of cranio-orbitofaciostenosis with hydrocephalus that has to date been associated with a uniformly poor outcome and frequent death in infancy. Assessment of the primary deformity and the approach to treatment in a consecutive series of 10 patients with the cloverleaf skull anomaly in the one unit are presented. Early predictable surgical correction of the cranio-stenosis is possible by fronto orbital advancement and lambdoid craniectomy. Less satisfactory correction of hydrocephalus and orbitostenosis is possible with the attendant increased morbidity. Early survival and the potential for a satisfactory long-term outcome depend on the management of the faciostenosis and an airway that may be anomalous at multiple levels. The initial approach was routine tracheostomy, which provided short-term relief but with the morbidity associated with such airway maintenance in any environment other than the most sophisticated health services. More recently, upper airway surgery (uvulopalatopharyngoplasty, adenoidectomy, and soft palatal split) has ensured airway control and avoided the progression to tracheostomy. When utilized later (1 to 10 years), such surgery may alleviate the symptoms of upper airway obstruction and sleep apnea and permit delay of midfacial advancement. PMID- 8416515 TI - Use of the osteocutaneous free scapular flap on the lower extremities. AB - Reports on the use of osteocutaneous free scapular flap transfers have focused primarily on mandible reconstruction. Yet the scapula is a nearly straight bone, and it is also useful in the reconstruction of long bones. We would like to report on 23 patients undergoing osteocutaneous free scapular flap transfers to the lower extremities in whom we obtained extremely satisfactory results. These 23 surgical patients included 10 with pseudoarthrosis with osteomyelitis, 3 with pseudoarthrosis due to trauma, and 5 with osteomyelitis of the lower extremities, as well as 5 with other miscellaneous problems. Surgery was successful in all patients. There was a recurrence of osteomyelitis in 2 patients. In 1 patient, the problem was minor and was treated on an outpatient basis. In the other patient, an infection arose in surgical dead space, and a second operation was required. The level of weight bearing achieved with the osteocutaneous free scapular flap is the same as obtained with a vascularized free fibular graft used for the treatment of pseudoarthrosis. We therefore believe that the osteocutaneous free scapular flap is extremely effective in the treatment of chronic and severe lower leg problems. PMID- 8416516 TI - Extracorporeal circulation for tissue transplantation (in the case of venous flaps). AB - During free-tissue transplantation, recipient vessels are often crushed as a result of trauma or radiation therapy, making anastomosis of vessels difficult. In some cases, a second operation is unavoidable because of formation of a postoperative thrombus at sites of the anastomosis. To solve such problems, it should be possible to transplant tissue anywhere in the body without vascular anastomosis by extracorporeal circulation. This study mainly concerned venous flaps that were spindle-shaped and 4 x 3 cm in size. One hundred and forty-six male Japanese White rabbits, weighing 2.0 to 2.5 kg, were used. Stabilized hemoglobin, lactated Ringer's solution, whole blood, and plasma were used as perfusate. Each of these perfusates was injected at a rate of 5 ml per hour continuously for 3 days in good recipient sites and for 7 days in poor recipient sites. Five rabbits were used in each group. The flaps were examined histologically and electron microscopically. Moreover, in the plasma group, microangiography was performed to observe blood circulation between the flap and the surrounding tissue. The flaps survived in all cases in which the plasma group had good recipient beds but did not survive in any of the other cases, in which degeneration of the endothelial cells was detected prior to edema. According to the results of this examination, tissue viability of the venous flap could be maintained for at least 3 days by perfusion of plasma. During this time, in the cases of good recipient sites, blood may begin to recirculate between the flap and the surrounding tissue. PMID- 8416517 TI - Blood flow and tissue survival in the rabbit venous flap. AB - The extent of venous flow, revascularization, local fluid imbibition, and metabolic status was evaluated in an experimental venous flap model. Thirty-six rabbits divided into six groups of six rabbits each had a 3.5 x 2.5 cm venous flap elevated along the thoracoepigastric vein, connected only by its proximal and distal vein, and sutured back. A composite graft of the same size was created on the contralateral side. Venous flaps survived 14 days, while composite grafts consistently did not. The vascular network was partially filled with fluorescein tracer within an hour after flap creation, even with an underlying Silastic sheet. Filling improved over several days, consistent with rapid revascularization. Composite grafts showed no immediate filling and delayed revascularization. Venous flow was apparently insufficient to enhance metabolism, since both glucose and lactate levels were equivalent between venous flaps and composite grafts. This supports the concept that an enhanced revascularization may be the primary mechanism of survival for venous flaps. PMID- 8416518 TI - Learning from the great masters. PMID- 8416519 TI - Changes and choices. PMID- 8416520 TI - Meeting mania. PMID- 8416521 TI - A rare case of arhinia with severe airway obstruction: case report and review of the literature. AB - A rare case of a newborn suffering from arhinia with complete airway obstruction is reported. The complexity of the life-threatening airway obstruction is described and adequate treatment is reported. The relevant literature is reviewed, and the different approaches to treatment are discussed. PMID- 8416522 TI - Congenital skin fossae in the zygomatic region. AB - A unique case of a congenital skin fossa in the zygomatic region in a 3-year-old girl is reported. Little has been written about congenital fossae, or dimples. They are thought to develop in the wound resulting from the fetal tissue being compressed between a sharp bony point and the uterine wall. The skin and subcutaneous tissue become compressed and adherent, and when the pressure is released, surrounding parts can stand up, while the attached part remains tied down, forming small dimples or fossae, what have been called "pressure dimples." This is the first report of a skin fossa located in the zygomatic region, as far as we know. PMID- 8416523 TI - Clam nail deformity of the little finger. AB - A case of a 2-month-old infant who had an extremely rare congenital nail deformity of the right little finger was reported. The nail looked like a clam, wrapping three-quarters of the distal phalanx. A neurovascular pedicle island flap was transposed to reconstruct the pulp and lateral nail fold with good results. Early surgery is justifiable to avoid growth disturbance of the fingertip. PMID- 8416524 TI - Lip augmentation for correction of thin lips. AB - Lip augmentation using a specially designed double V-Y advancement flap has been performed on eight patients, and a total of 14 lips were done. The results in the patients who are now over 1 year postoperative are encouraging as a permanent solution to augment thin lips. Relatively long postoperative swelling and stiffness have been a drawback and require careful patient selection. PMID- 8416525 TI - Refinements in pterygomaxillary dissociation for total midface osteotomies: instrumentation, technique, and CT scan analysis. PMID- 8416526 TI - Inverted running interlocking suture (IRIS): a technique for rapid closure of wounds. AB - I have used the IRIS technique during the past several years on a number of patients. Impressive time saving, simplicity of technique, and excellent tissue approximation have been observed. The technique also provides transverse and vertical immobilization of the wound, a favorable condition for better healing. PMID- 8416527 TI - Evaluating a plastic surgery academic faculty position. PMID- 8416528 TI - International commemoration of the 200th birthday of Johann Friedrich Dieffenbach, February 1-2, 1992 in Berlin. PMID- 8416529 TI - Dermatolipectomy of the lower thigh. PMID- 8416530 TI - Flumazenil: a new benzodiazepine antagonist. PMID- 8416531 TI - Infection of Proplast malar implants and dental injections. PMID- 8416532 TI - Symbrachydactyly instead of atypical cleft hand. PMID- 8416533 TI - Hypnotic doses of propofol block ketamine-induced hallucinations. PMID- 8416534 TI - A simple method to avoid depressed scarring in laser treatment of elevated lesions. PMID- 8416535 TI - Ultrasound and seromas. PMID- 8416536 TI - Complications of prolonged bilateral lower extremity tourniquet. PMID- 8416537 TI - Reconstruction of the bone--bone marrow organ by osteogenin, a bone morphogenetic protein, and demineralized bone matrix in calvarial defects of adult primates. AB - Information concerning the efficacy of osteogenin, a bone morphogenetic protein, and demineralized bone matrix in orthotopic sites in nonhuman primates is a prerequisite for potential clinical application in humans. After exposure of the calvaria, 84 cranial defects, 25 mm in diameter, were prepared in 26 adult male baboons (Papio ursinus). Defects were implanted with insoluble collagenous bone matrix (ICBM, the inactive collagenous residue after dissociative extraction of bone matrix with 4 M guanidine hydrochloride) reconstituted with osteogenin fractions isolated from baboon bone matrix by chromatography on heparin-Sepharose and hydroxyapatite-Ultrogel (Og Hep-HA) or osteogenin further purified using Sephacryl S-200 gel filtration chromatography (Og S-200). Baboon osteogenin with the highest biologic activity in a rodent bioassay, as determined by alkaline phosphatase activity, calcium content, and histologic analysis, was used for orthotopic implantation in baboons. Additional defects were implanted with baboon demineralized bone matrix (DBM) or ICBM without osteogenin as control. Defects also were grafted with corticocancellous bone harvested from the iliac crest or left ungrafted to monitor the spontaneous regeneration potential of the adult baboon calvaria. Undecalcified bone sections at 7 microns were prepared from the harvested specimens 30 and 90 days after surgery. Histomorphometry demonstrated that Og S-200 induced copious amounts of bone and osteoid as early as day 30 (P < 0.01 versus ICBM, autogenous grafts and untreated defects). At day 90, in implants of Og S-200, Og Hep-HA, and DBM, bone and marrow formation was extensive, culminating in complete regeneration of the craniotomies. In implants of DBM, bone formed with an intervening phase of cartilage development. This provides the phenotypic evidence of endochondral bone differentiation by induction in defects of membranous calvarial bone in adult primates. These results establish the potential therapeutic application of osteogenin and demineralized bone matrix for the architectural reconstruction of the bone-bone marrow organ in humans. PMID- 8416538 TI - The prolabial unwinding flap method for one-stage repair of bilateral cleft lip, nose, and alveolus. AB - This paper describes a one-stage reconstruction of the complete bilateral cleft lip, nose, and alveolus using an asymmetrically designed prolabial flap. The columella of the nose and the central lip are produced by "unwinding" the columellar and labial sections of the prolabium around a small central tab, which is used to center the junction between the lip and columella. Only two scars are produced, both of which run vertically along each philtral column. Accurate preoperative orthopedics without lip adhesion is considered essential to this repair. The principal advantage/disadvantage to the method is its asymmetrical design. If asymmetries result, they have been easily corrected. Eight consecutive cases are presented with follow-up ranging from 1.5 to 6.3 years. PMID- 8416539 TI - Tip graft: a 20-year retrospective. AB - This paper chronicles a personal experience with nasal tip grafts over 20 years. In the first period (1968-1975), the original graft was designed for use in secondary rhinoplasty cases to obtain both projection of the tip and increased angulation at the columellar-lobular junction. The use was soon expanded to primary patients with inadequate tip projection. During the middle period (1975 1982), the applications for tip grafting were expanded to include many kinds of tip problems (both primary and secondary), cleft lip noses, and various ethnic noses, especially those with thick skin. The incidence of postoperative displacement and/or visibility of the graft was reduced as multiple grafts of solid, bruised, and crushed cartilage became routine. Ear cartilage was first used and ethmoid was abandoned as a primary graft. The incidence of infection was significantly reduced. The current period (1982-1991) is marked by refinements in technique and materials. Greater versatility with graft composition and materials makes possible a variety of tip configurations, custom-made for individual requirements. PMID- 8416540 TI - Risk factors for infection following operative treatment of mandibular fractures: a multivariate analysis. AB - Utilizing a retrospective study design and a study sample of 284 consecutive patients, we measured the association between five different risk factors and the development of infection following the operative management of mandibular fractures. The five risk factors analyzed were age, sex, number of fractures per patient, time from admission to treatment, and treatment modality. The overall infection rate was 5.3 percent. The infection rate for each treatment modality was (1) 0 percent for closed reduction, (2) 20.0 percent for open reduction and internal fixation with wire osteosynthesis, and (3) 6.3 percent for open reduction and rigid internal fixation. To measure the association between the various risk factors and postoperative infection, we used multivariate logistic regression. After controlling for potential confounding variables, the only risk factor statistically associated with postoperative infection was treatment modality. PMID- 8416541 TI - Successful pregnancies following TRAM flap surgery. PMID- 8416542 TI - Expansion of fascial flaps: histopathologic changes and clinical benefits. AB - In order to meet the requirements of large defects created by sacral and trochanteric pressure ulcers, tensor fasciae latae and lumbosacral fasciocutaneous flaps were expanded with a silicone expander in six patients (10 flaps). In 8 of the flaps the location of pressure ulcers was trochanteric, and in 2 it was sacral. In each case the period of expansion was 4 weeks. Histopathologic examination after expansion showed an increase in the vascularity and overall thickness of fascia in both types of flaps. After 4 weeks of expansion, the perifascial areolar tissue was replaced with thick granulation tissue in the tensor fasciae latae flaps. This markedly vascular layer of granulation tissue interposed between the fasciae and the subcutaneous tissue augmented the internal matrix of the flap, thereby lowering the potential for shearing during flap elevation. Conceivably, owing to the absence of an areolar tissue layer in the lumbosacral fasciocutaneous flaps, no layer of granulation tissue was observed. The average thickness of the fibrous capsule formed around the expanders was 573.2 microns, which was composed of three structurally different zones. Prior tissue expansion obviously assisted primary closure of the flap donor site. In addition, it seems that the tissue-expansion process rendered the distal portion of fascial flaps more robust because of increased vascularity. It is therefore proposed that preparatory tissue expansion of fascial flaps has several advantages. The obvious benefits include the ability to close larger defects while closing the donor site primarily. As a result of this study, the additional benefits may include a reduction in the mechanical shear potential of these flaps and an improvement in their vascularity. PMID- 8416543 TI - Cyclosporine for pyoderma gangrenosum. AB - A case of pyoderma gangrenosum that occurred in the absence of underlying illness and was initially misdiagnosed is described. Hemorrhagic pustules and ulcers appeared over vein-graft sites after open heart surgery and were treated with cyclosporine. The result was rapid and complete resolution. It is the purpose of this report to emphasize pyoderma gangrenosum as a cause of ulceration in healthy individuals and to highlight the addition of cyclosporine to the therapeutic armamentarium. PMID- 8416544 TI - Data on first recurrence after treatment for malignant melanoma in a large patient population. AB - Recurrent malignant melanoma is a clinical entity that behaves distinctly differently from the primary disease. The purpose of this study was to determine whether there are significant trends in the prognostic factors or in the timing and sites of recurrence. We reviewed 250 patients with recurrent malignant melanoma from the Sydney Melanoma Unit in Australia who were followed in their Tumor Registry between 1960 and 1981 (median follow-up was 7 years). Fifty-two percent of all first recurrences were to regional nodes, 17 percent were local recurrences, 8 percent were in-transit metastases, and 23 percent of all first recurrences were to a distant organ. We found that 67 percent of all recurrences were diagnosed within 24 months and 81 percent were diagnosed by 36 months after the primary melanoma. Regional nodal recurrences were diagnosed earlier (with 63 percent at 18 months and 74 percent at 24 months). Local recurrences and in transit metastases were comparable (with 55 percent of each at 18 months and about 66 percent of each at 24 months), and systemic recurrences were diagnosed later (52 percent at 24 months and 71 percent at 36 months). Of significant interest was the fact that survival after the diagnosis of recurrence was independent of thickness of the primary tumor or duration of disease-free interval (local, in-transit, or regional nodal). Only the diagnosis of distant organ metastasis correlated with shorter survival; local recurrence, in-transit metastasis, and regional nodal metastasis were all associated with comparable survivals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416545 TI - Microvascular treatment of impotence. AB - Occlusion and/or stenosis of arteries supplying the penis was the cause of erectile impotence in 11 patients. They were chosen from 124 patients who were assessed by a multidisciplinary team for impotence. Indirect revascularization of the corpora cavernosa was performed by anastomosing the inferior epigastric artery to the deep dorsal penile vein. This arterialized vein was, in turn, anastomosed side to side to one of the corpora cavernosa. In seven patients the anastomosis remained patent. There was one documented thrombosis. Three patients were unavailable for Doppler examination but are known to have achieved erection. The indirect revascularization of the corpora cavernosa appears to be an acceptable surgical treatment of vasculogenic impotence. PMID- 8416546 TI - Gradient-echo MR imaging: techniques and acronyms. AB - This overview briefly traces the history and nomenclature of gradient-recalled echo (GRE) techniques used in magnetic resonance (MR) imaging. GRE sequences, which are now offered commercially by 11 major manufacturers of MR imagers, are presented to illustrate their structural similarities and subtle differences in implementation. A classification scheme is proposed for these sequences employing a vendor-independent nomenclature. A glossary of GRE abbreviations and acronyms found in the current MR marketplace also is presented. PMID- 8416547 TI - Epidermal inclusion cyst after reduction mammoplasty. AB - Reduction mammoplasty has become an increasingly common surgical procedure in the United States. Because the technique generally involves repositioning of the nipple-areolar unit with infolding of a vascularized tissue pedicle, epidermal inclusion cysts may develop after the procedure. The authors encountered three cases of epidermal inclusion cysts after reduction mammoplasty and describe the associated mammographic and sonographic characteristics. The authors found that stereotaxic needle biopsy is useful for the diagnosis of these lesions. PMID- 8416548 TI - Morphologic effects of cigarette smoking on airways and pulmonary parenchyma in healthy adult volunteers: CT evaluation and correlation with pulmonary function tests. AB - A prospective computed tomographic (CT) study was performed to determine the prevalence of lung changes in smokers. The study group comprised 175 healthy adult volunteers (current smokers, n = 98; ex-smokers, n = 26; nonsmokers, n = 51). The subjects underwent clinical examination, pulmonary function tests, chest radiography, and conventional and high-resolution CT (HRCT). Significant differences between current smokers, ex-smokers, and nonsmokers were observed with HRCT in the identification of subpleural (P = .11) and parenchymal (P < .001) micronodules, emphysema (P < .001), and areas of ground-glass attenuation (P = .0001). All subjects had normal pulmonary function. Parenchymal micronodules, areas of ground-glass attenuation, and emphysema were observed with a significant predominance in the upper lung zones (P < .01). Presence of emphysema and abnormal bronchial wall thickening were the only HRCT signs associated with significantly lower values of functional parameters. These data support the concept that parenchymal abnormalities can be detected in healthy smokers with normal findings at chest radiography and pulmonary function tests. PMID- 8416549 TI - Expectant management of ectopic pregnancy. PMID- 8416550 TI - Dynamic pulmonary CT: findings in healthy adult men. AB - The authors examined 10 healthy male subjects with dynamic computed tomography to determine normal dynamic and expiratory findings. In both the supine and prone positions, the average increase in lung attenuation during exhalation was found to be significantly greater in dependent lung regions than in nondependent lung regions. In each of the lung zones studied, there was a significant correlation between the decrease in cross-sectional lung area during exhalation and the increase in lung attenuation. At the lung bases, a greater increase in lung attenuation was noted during exhalation for a given change in cross-sectional lung area than in the upper lungs. All 10 subjects showed a distinct lobar attenuation gradient in the supine position. In general, lung attenuation increased homogeneously during exhalation, but four of the 10 subjects showed regions of inhomogeneity in lung attenuation during rapid exhalation indicative of air trapping, despite normal results on pulmonary function tests. PMID- 8416551 TI - Pulmonary talcosis: CT findings in three cases. AB - The authors describe the computed tomographic (CT) appearances in three patients with pulmonary talcosis resulting from chronic intravenous drug abuse. There was widespread ground-glass attenuation in one case and an appearance similar to that of progressive massive fibrosis in two cases. In the latter cases, there were confluent perihilar masses with areas of high attenuation. While the CT appearances may be suggestive of pulmonary talcosis, tissue sampling is required for definitive diagnosis. PMID- 8416552 TI - T1 lung cancer: prevalence of mediastinal nodal metastases and diagnostic accuracy of CT. AB - To determine the prevalence of mediastinal lymph node metastases in T1 non-small cell lung cancer and assess the sensitivity and specificity of computed tomography (CT) in detection of such metastases, the CT scans and surgical findings in 104 patients with T1 lesions were reviewed. Nodes longer than 10 mm on the short or long axis were considered abnormal. All patients underwent thorough mediastinal lymph node dissection at mediastinoscopy or thoracotomy. A total of 362 lymph nodes were sampled. Nodal metastases were present in 22 patients (21%). The sensitivity of CT for metastases to individual nodal stations was 41% for nodes measured on the short axis and 55% for those measured on the long axis. The specificity was 93% and 86%, respectively. When the adjacent nodal stations were included in the analysis, the sensitivity of CT was 59% for nodes measured on the short axis and 77% for those measured on the long axis; the specificity was 91% and 73%, respectively. T1 lung cancer has a higher prevalence of lymph node metastasis than previously reported, and CT is recommended in the preoperative staging of this disease. PMID- 8416553 TI - Clinical PET: Aesop's tortoise? PMID- 8416554 TI - Hypervascular liver lesions: differentiation of focal nodular hyperplasia from malignant tumors with dynamic gadolinium-enhanced MR imaging. AB - The differentiating points between focal nodular hyperplasia (FNH) and malignant hypervascular liver lesions were studied at dynamic gadolinium-enhanced magnetic resonance (MR) imaging. Thirty-six patients with 50 hypervascular lesions (28 FNH, 12 hepatocellular carcinoma, nine metastases, and one cholangiocarcinoma) underwent unenhanced spin-echo (SE) T1- and T2-weighted imaging and T1-weighted gradient-recalled-echo imaging before and repeatedly for 10 minutes after intravenous bolus injection of gadopentetate dimeglumine. On unenhanced SE images, the signal intensity of 25 FNH lesions (89%) and 10 malignant tumors (45%) was homogeneous. A central scar was detected in 12 FNH lesions (43%) and in none of the malignant tumors. On dynamic gadolinium-enhanced images, all lesions had early vigorous enhancement that was homogeneous in 27 FNH lesions (96%) and in seven malignant tumors (32%) (P < .001). After administration of gadopentetate dimeglumine, central scars were seen in 22 FNH lesions (79%) and in one malignant tumor (4%) (P < .001). All FNH lesions (100%) and six malignant tumors (27%) had well-defined enhancement (P < .001). There was overlap in the enhancement pattern between hypervascular malignant lesions and FNH, but by using the combination of unenhanced and enhanced images, they could be distinguished. PMID- 8416555 TI - Acute liver rejection: evaluation with cell-directed MR contrast agents in a rat transplantation model. AB - Enhancement patterns with gadolinium (4s)-4-(4-ethoxybenzyl)-3,6,9-tris (carboxylatomethyl)-3,6,9-triazaundecandioic acid (EOB) diethylenetriaminepentaacetic acid (DTPA), a hepatocyte-directed magnetic resonance (MR) contrast agent, and superparamagnetic iron oxide particles (SPIO), a Kupffer cell-directed contrast agent, were compared at MR imaging in rat models of allogeneic and syngeneic liver transplantation. Light and electron microscopy were performed to monitor the morphologic changes in these livers and revealed typical findings in acute rejecting liver (allogeneic transplants) and normal architecture in syngeneic transplants. Liver enhancement with Gd-EOB-DTPA at MR imaging was delayed and prolonged with transplantation, and even more with rejection, but the level of enhancement when compared with that of control rats remained unaffected. Uptake of SPIO was decreased in the acutely rejecting livers but was normal in the syngeneic transplantation model. Enhanced MR imaging with cell-directed contrast agents depicted decreased phagocytotic activity in acute transplant rejection and enabled the excretory function of hepatocytes to be monitored. Additionally, Gd-EOB-DTPA may be applied advantageously for detection of focal lesions in transplanted livers. PMID- 8416556 TI - Three-dimensional spiral CT angiography of the abdomen: initial clinical experience. AB - Spiral computed tomography (CT) is a new technology that couples continuous tube rotation with continuous table feed. This allows compilation of a data set that has continuous anatomic information without the establishment of arbitrary boundaries at section interfaces as in conventional CT. The unique method of data collection of the spiral scanner has been combined with a dynamic intravenous contrast material bolus to image abdominal vasculature, specifically, the aorta, renal arteries, and splanchnic circulation. Through various techniques of image processing, including surface renderings and maximum-intensity projections, it is possible to obtain excellent anatomic detail of the aorta and its major branches. The authors applied this technique in 15 patients and reliably saw third-order aortic branches as well as third-order splenic-portal venous anatomic detail with remarkable clarity. Pathologic conditions detected include stenotic renal arteries, abdominal aortic dissection, abdominal aortic aneurysm, and celiac bypass graft occlusion. PMID- 8416557 TI - Evaluation of the prostate and prostatic carcinoma with gadolinium-enhanced endorectal coil MR imaging. AB - Thirteen patients who underwent subsequent radical prostatectomy for prostate cancer were studied with spin-density-T2-weighted and gadolinium-enhanced and unenhanced T1-weighted magnetic resonance (MR) imaging performed with an endorectal surface coil. With gadopentetate dimeglumine, the central gland showed inhomogeneous enhancement due to benign hyperplasia, whereas the peripheral zone demonstrated more uniform and less intense enhancement. With the T1-weighted sequence, depiction of the zonal anatomy, prostate capsule, surgical capsule, fibromuscular stroma, and periprostatic venous plexus was better with contrast enhancement, but these were best demonstrated on T2-weighted images. The trend was similar for prostatic tumor extent and capsular integrity. The seminal vesicles had a septal enhancement pattern and in several patients were best evaluated with enhanced T1-weighted images. These findings suggest that gadopentetate dimeglumine is not warranted for routine use in endorectal MR imaging of the prostate but may be useful for evaluation of the seminal vesicles in selected patients. PMID- 8416558 TI - Zonal anatomy of the cervix: correlation of MR imaging and histologic examination of hysterectomy specimens. AB - High-resolution magnetic resonance (MR) images of nine hysterectomy specimens were correlated with light microscopy, computer-aided image analysis, and immunohistochemical examination to investigate the histologic counterparts of the zonal anatomy of the cervix. A central stripe of high signal intensity, a surrounding middle layer of low signal intensity, and an outer layer of intermediate signal intensity were found in all specimens. Histologic correlation indicated that the central stripe most likely represents the secretions in the canal, the cervical mucosa, and the plicae palmatae; the other two layers represent fibromuscular stroma. The percentage of nuclear area in the inner zone of the fibromuscular stroma is 2.5 times greater than in the outer zone, which may account for the lower signal intensity of the inner zone. No difference in distribution of collagen, laminin, and fibronectin (common components of the extracellular matrix) was found between the two zones of the cervical fibromuscular stroma. PMID- 8416559 TI - Malignant ureteral obstruction: preliminary results of treatment with metallic self-expandable stents. AB - Self-expandable metal stents 7 mm in diameter were percutaneously implanted into 13 ureters in 10 patients with malignant ureteral obstruction not amenable to double-J stent placement. In nine ureters, one stent was placed, and in four ureters, two overlapping stents were placed. Primary reconstitution of ureteral patency was achieved in all ureters. After 1-2 weeks, four ureters showed a urothelial reaction encroaching on the lumen of the ureter, and a double-J stent was placed coaxially. One ureter was occluded by urothelial hyperplasia 4 weeks after stent placement, and a double-J stent was therefore placed. One ureter was occluded 8 months after stent placement by distal tumor overgrowth. The other ureters showed no signs of obstruction during a follow-up of 3-14 (average, 5.8) months. Peristalsis was preserved at both ends of the stent in all ureters. The use of this stent alone or in combination with a double-J stent alleviated upper urinary tract obstruction and avoided external drainage in all patients. PMID- 8416560 TI - Dual mechanisms for change in myocardial signal intensity by means of a single MR contrast medium: dependence on concentration and pulse sequence. AB - To determine whether gadodiamide injection can provide sufficient enhancement on both T1- and T2-weighted spin-echo magnetic resonance (MR) images of the heart and skeletal muscles, anesthetized rats were divided into five groups. Groups 1-3 received 0.1 (n = 9), 0.3 (n = 8), or 0.5 (n = 8) mmol/kg gadodiamide injection, respectively, and T1-weighted images were obtained. Groups 4 and 5 received 0.3 or 0.5 mmol/kg gadodiamide injection, respectively, and T2-weighted images were obtained. Gadolinium concentration was measured in myocardium by means of inductively coupled plasma-atomic emission spectroscopy. On T1-weighted images, the 0.1 and 0.3 mmol/kg doses produced a dose-dependent increase in myocardial signal intensity proportional to gadolinium concentration. A dose of 0.5 mmol/kg, which correlated with higher gadolinium concentration and did not further increase myocardial signal intensity, prolonged the imaging window. On T2 weighted images, the 0.3 mmol/kg dose caused a transient decrease in myocardial signal intensity; the 0.5 mmol/kg dose produced greater and persistent loss of signal intensity. In conclusion, the changes in signal intensity induced by gadodiamide injection depend on the dose, pulse sequence, and type of tissue. PMID- 8416561 TI - Nephrotoxicity of nonionic low-osmolality versus ionic high-osmolality contrast media: a prospective double-blind randomized comparison in human beings. AB - To test whether a nonionic, low-osmolality contrast medium (iopamidol) administered for coronary angiography was less harmful to renal function than ionic, high-osmolality medium (sodium diatrizoate), a prospective, double-blind randomized study of 70 patients with normal or mildly depressed renal function (serum creatinine < or = 2.0 mg/dL (175 mumol/L) was performed. Creatinine clearance was determined before coronary angiography and 24 and 48 hours after. There were no significant differences between the low- and high-osmolality groups with regard to age, baseline creatinine clearance, or dose of contrast medium given. In patients receiving low-osmolality medium (n = 35), creatinine clearance decreased by 19% +/- 13 (1 standard deviation) at 24 hours and recovered by 48 hours. In patients receiving high-osmolality medium (n = 35), creatinine clearance decreased by 40% +/- 16 at 24 hours and remained depressed by 47% +/- 14 at 48 hours. In patients with normal or mildly depressed renal function, use of a non-ionic, low-osmolality contrast medium minimized nephrotoxicity as measured by reductions in creatinine clearance after coronary angiography. PMID- 8416562 TI - Aortic regurgitation: quantitation with MR imaging velocity mapping. AB - Aortic regurgitation (AR) in five healthy volunteers and 26 patients (mean age, 60.3 years; range, 25-83 years) was quantitatively measured with magnetic resonance (MR) imaging velocity mapping. Cine transverse images of the ascending aorta (32 phases per cardiac cycle) were acquired by using a gradient-echo sequence with a velocity-encoding bipolar pulse applied in the section-selection direction with a 1.5-T MR imaging unit. The aortic flow was calculated by integrating the product of area and mean velocity of the ascending aorta at each phase over a cardiac cycle. The negative and positive velocity values indicated antegrade and regurgitant flow, respectively, which allowed calculation of forward and regurgitant flow. Inter- and intraobserver variation of regurgitant fraction (RF) measurement was small (r = .956, standard error of the estimate [SEE] = 1.2%, n = 31; and r = .998, SEE = 0.35%, n = 10, respectively). RF determined with MR imaging agreed well with Doppler echocardiographic (n = 26) and aortographic (n = 9) grading of AR. Reproducible, quantitative, and noninvasive measurement of AR is possible with MR velocity mapping. PMID- 8416563 TI - Pneumothorax: detection with upright versus decubitus radiography. AB - To evaluate the value of decubitus chest radiography in the clinical assessment of pneumothorax, the authors compared the detectability of pneumothorax on expiratory, upright chest radiographs with that on expiratory, lateral decubitus radiographs obtained with the suspected side up. Five radiologists reviewed 19 sets of radiographs with pneumothoraces and 20 sets of radiographs without pneumothoraces. All five radiologists were more accurate and had greater confidence in the evaluation of the upright studies. A pneumothorax was detected 21% (77 versus 57 of 95) more often on the erect radiographs. In three cases, however, decubitus positioning was more definitive. These results suggest that when clinically feasible, the expiratory, upright chest radiograph is the procedure of choice for the evaluation of small pneumothoraces. PMID- 8416564 TI - Aortic aneurysm and dissection: normal MR imaging and CT findings after surgical repair with the continuous-suture graft-inclusion technique. AB - The normal range of postoperative imaging findings are described in 34 asymptomatic patients studied 5-66 months (mean, 28 months) after undergoing the continuous-suture graft-inclusion technique for repair of aortic aneurysms (n = 20) and dissections (n = 14) involving the ascending aorta. All 34 patients underwent magnetic resonance (MR) imaging, and 24 patients also underwent computed tomography (CT). Perigraft thickening was seen in 19 patients (56%) with MR imaging and in eight patients (33%) with CT. Flow outside the graft but contained within the native wrap was noted in five patients (15%) with MR imaging and in four patients (17%) with contrast material-enhanced CT. Thrombus was identified outside the graft and within the wrap in seven patients (21%) with MR imaging and in six patients (25%) with CT. Mass effect on the graft was depicted in four patients (12%) with MR imaging and in three patients (13%) with CT. Of the 14 patients who underwent repair of aortic dissections, an intimal flap was seen distal to the graft in seven of the 14 (50%) evaluated with MR imaging and in four of the 10 (40%) evaluated with contrast-enhanced CT. An accurate postoperative imaging evaluation requires precise knowledge of the surgical technique performed and its anatomic consequences. PMID- 8416565 TI - Intraportal US with 20-MHz and 30-MHz scanning catheters. Work in progress. AB - Intraportal ultrasonography (US) with scanning catheters was attempted in nine patients with various biliary tract diseases, to evaluate the portal venous system for tumor invasion. Intraportal US was performed successfully in seven patients. Intraportal US scans were compared with images obtained with arterial portography, direct portography, and intraoperative US. Intraportal and intraoperative US revealed cancer invasion into the wall of the portal venous trunk in two patients. Negative findings for cancer invasion with intraportal US were verified with intraoperative US and laparoscopic examination in five cases. Arterial portography was unable to delineate the fine portal venous wall configuration in six of the seven patients, and direct portographic images were not fully diagnostic of tumor invasion in three cases. The improved detection of tumor invasion to the portal venous system with intraportal US was helpful in determining the appropriate treatment for biliary tract disease in these patients. PMID- 8416566 TI - Iliac arteries: reanalysis of results of balloon angioplasty. AB - In this reanalysis of the 667 percutaneous transluminal balloon angioplasties (PTAs) of the iliac arteries, the variables that are predictive of early and late success were identified. For 82 iliac occlusions, the success rate at 1 month was 75.6% +/- 4.7 (mean +/- standard error). With exclusion of the 15 technical failures that were associated with one complication, the success rate at 1 month was 91.0% +/- 3.5 and at 3 years, 58.8% +/- 7.1. For PTA of an iliac occlusion at one site, the predicted 3-year success rate was 66%, whereas for PTA of an iliac occlusion with a tandem lesion, the success rate was 17%. For the 313 common iliac PTAs, the success rate at 1 month was 97.1% +/- 0.9; at 1 year, 81.1% +/- 2.3; at 2 years, 70.6% +/- 2.9; at 3 years, 67.8% +/- 3.0; at 4 years, 64.9% +/- 3.3; at 5 years, 60.2% +/- 4.0; and at 6 years, 52.0% +/- 5.7. There were no variables that were statistically predictive of late results. For the 209 external iliac PTAs, the predicted 3-year success rate was 57% for men and 34% for women. For 58 PTAs of both common and external iliac stenoses, the predicted 3-year success rate was 73% for patients with good runoff and 30% for those with poor runoff. Serious complications were recorded in 3.9% (26 of 667): Death occurred in 0.3%, operation was necessary in 1.0%, and hospital discharge was delayed in 2.6%. PMID- 8416567 TI - Use of preprocedural tests by interventional radiologists. AB - A direct survey was taken of 1,238 members of the Society of Cardiovascular and Interventional Radiology (SCVIR) concerning the routine use of 14 tests (most frequently, prothrombin and partial thromboplastin times, complete blood cell counts, and blood urea nitrogen and serum creatinine levels) performed before 10 invasive percutaneous procedures (peripheral angiography, neuroangiography, transluminal angioplasty, thrombolysis, percutaneous needle biopsy, abscess drainage, percutaneous nephrostomy, biliary drainage, myelography, and venography). The survey was undertaken to determine the current practices and appropriateness of current routine use of preprocedural tests. The response rate was 34%, representing a cumulative annual volume of 322,208 cases. The practice of performing routine preprocedural tests is common among interventional radiologists. Data provided by this survey suggest that use of these tests is excessive. Adherence to suggested proposals derived from previously reported experience would result in an annual estimated savings of $20.0-$34.9 million (extrapolated for all procedures performed by SCVIR members in 1989). PMID- 8416568 TI - Reactive soft-tissue mass associated with osteoid osteoma: correlation of MR imaging features with pathologic findings. AB - The authors retrospectively reviewed three cases of histologically documented osteoid osteoma in which magnetic resonance (MR) imaging was performed prior to surgical excision. In all three cases, MR imaging demonstrated abnormal signal intensity characteristics, and use of gadopentetate dimeglumine resulted in enhancement. These findings correlated with a reactive soft-tissue mass with myxomatous change, as well as with cell-depleted, juxtanidal bone marrow that contained proteinaceous material. These MR imaging findings can easily be confused with those of a malignant tumor or osteomyelitis. PMID- 8416569 TI - Osteoid osteomas of the femoral neck: report of four cases evaluated with isotopic bone scanning, CT, and MR imaging. AB - Four patients, ranging in age from 6 to 32 years, with surgically proved parosteal lesions of the femoral neck are presented. In one case, magnetic resonance (MR) imaging was the initial study ordered prior to plain radiography, and in three cases MR imaging was performed after plain radiography. Findings in the MR images were abnormal in all four cases; however, the nidus was not identified prospectively. Incorrect diagnoses based on findings at MR imaging of secondary marrow edema and/or synovitis included Ewing tumor, osteonecrosis, stress fracture, Lyme arthritis, and juvenile inflammatory arthritis. In all four cases, the correct diagnosis of intracapsular osteoid osteoma was made following (a) review of plain radiographs leading to a high index of suspicion and (b) performance of thin-section computed tomographic (CT) studies (in one case a screening CT study with 1-cm-thick sections failed to depict the lesion). In three cases, isotopic bone scans provided useful guidance for planning the CT examination. PMID- 8416570 TI - Pneumothorax detection: a problem in experimental design. PMID- 8416571 TI - MR imaging of the tarsal sinus and canal: normal anatomy, pathologic findings, and features of the sinus tarsi syndrome. AB - After definition of the normal anatomic features of the tarsal sinus and canal at magnetic resonance (MR) imaging, 123 ankle MR imaging studies in 116 patients were reviewed. Abnormalities of the tarsal sinus and canal were seen on MR images in 33 cases (26.8%), were highly associated with tears of the lateral collateral ligament, and could be categorized according to the pathologic findings in patients with sinus tarsi syndrome: (a) diffuse infiltration with low T1- and T2 weighted signal intensity (n = 17) consistent with fibrosis, (b) diffuse infiltration with low T1-weighted signal intensity and increased T2-weighted signal intensity (n = 11) consistent with chronic synovitis and nonspecific inflammatory changes, and (c) multiple abnormal fluid collections (n = 5) consistent with synovial cysts. Absence of the anterior microrecesses of the posterior subtalar joint was a common finding on normal MR imaging studies (46 of 90) and may reflect lack of iatrogenic joint distention. Tears of the posterior tibial tendon may have a previously unrecognized association with the sinus tarsi syndrome. PMID- 8416572 TI - Vascularized iliac crest grafts: evaluation of viability status with marrow scintigraphy. AB - Forty-three vascularized iliac crest grafts in the femoral neck region in 38 patients were assessed with marrow scintigraphy. Twenty grafting procedures had been performed for treatment of fractures, 16 for nontraumatic osteonecrosis, and seven for focal bone lesions. Scintiscans were obtained preoperatively and in the early and late postoperative period. Anterior images of the hips were obtained 30 minutes after intravenous injection of 4 mCi (148 MBq) of technetium-99m tin colloid by using a gamma camera with a low-energy, general-purpose collimator. The viable graft was detected as an area of tracer uptake corresponding to the configuration of the graft on radiographs. Marrow scintigraphy at 12 weeks showed that 24 grafts were viable, 11 were nonviable, and eight were indeterminate. Early and late results concurred in 36 of 43 (84%) grafts. Nonviable grafts occurred most frequently in fractures in the elderly. In seven patients who subsequently required total hip replacement, six grafts were nonviable while one was indeterminate at 12 weeks. Results indicate that marrow scintigraphy is useful in the treatment of patients with vascularized iliac crest bone grafts. PMID- 8416573 TI - Atypical idiopathic scoliosis: MR imaging evaluation. AB - The authors analyzed the clinical and imaging findings in 30 consecutive pediatric, adolescent, and young adult patients who underwent MR imaging because of atypical features of idiopathic scoliosis. Atypical clinical and plain radiographic features included early onset (n = 4), rapid progression (n = 19), pain (n = 17), other neurologic symptoms or signs (n = 12), associated syndromes (n = 4), a convex left thoracic or thoracolumbar curve (n = 18), a kyphotic component (n = 7), and pedicle thinning (n = 4). MR imaging demonstrated 17 abnormalities in 11 patients: lumbar disk protrusions (n = 1), patulous intradural space (dural ectasia) (n = 3), hydrosyringomyelia (n = 7), Chiari I malformation (n = 5), and cord astrocytoma (n = 1). Significant associations with abnormal MR imaging findings were shown for patients with pain, weakness, abnormal neurologic findings, and atypical curvatures. Furthermore, there was a striking association of convex left thoracic or thoracolumbar scoliosis with hydrosyringomyelia (six of seven cases). On the basis of these results, "atypical" spinal curvatures at radiography and "atypical" clinical features should prompt performance of additional diagnostic studies. PMID- 8416574 TI - Diagnosis of hyperparathyroidism with US after autotransplantation: results of a prospective study. AB - The best treatment for reactive hyperparathyroidism, a complication of chronic renal failure, consists of resection of all four parathyroid glands and subsequent autotransplantation of parathyroid fragments into the brachioradialis muscle of the forearm. These procedures normalize the elevated parathyroid metabolism in at least two-thirds of the patients. In this prospective study, the parathyroid transplant was examined at least once with ultrasound (US) during postoperative follow-up for a maximum of 84 months in 22 women and 13 men who were undergoing long-term hemodialysis for reactive hyperparathyroidism. In 32 of these 35 patients, the graft-bearing forearm had a normal appearance on US scans. Three patients had one or two hypoechoic lesions 5-11 mm in diameter. Graft dependent recurrent hyperparathyroidism was verified in two of these patients by means of clinical, biochemical, and histologic examination. It is concluded that US is effective in the recognition and localization of graft-dependent recurrent hyperparathyroidism. PMID- 8416575 TI - Real-time system for angle-independent US of blood flow in two dimensions: initial results. AB - The authors developed an ultrasound system that enables the speckle patterns produced by echoes from moving blood to be tracked in real time. Unlike current color Doppler flow imagers, this system allows the measurement of blood velocities in any direction within the imaging plane. The authors used this device to image flow in the human jugular vein and contrasted the image with one obtained under similar circumstances with color Doppler flow imaging. The authors demonstrated that this system can display in vivo lateral blood flow in real time. Further development of the system, including the incorporation of wall filters to enhance weak blood echoes and parallel techniques to reduce data acquisition time, will allow clinical imaging of flow with velocities of several meters per second in any direction without aliasing or dependence on the Doppler angle. PMID- 8416576 TI - Receiver operating characteristic analysis of fracture and pneumonia detection: comparison of laser-digitized workstation images and conventional analog radiographs. AB - Forty pneumonia and 40 fracture cases were matched by patient age and gender with normal cases. Pediatric cases constituted 25% of this sample. All 160 examinations were laser-digitized at a spot size of 210 microns, a contrast resolution of 12 bits, and a spatial resolution of 2.35 line pairs per millimeter. The digitized images were transmitted 5 miles over a dedicated telephone line at a T1 rate from Francis Scott Key Hospital to Johns Hopkins Hospital (both in Baltimore, Md). At Johns Hopkins Hospital, eight radiologists interpreted the original analog radiograph and corresponding soft-copy display (1,280 x 1,024 pixels). Findings, confidence ratings, and image quality were reported for each reading. A receiver operating characteristic (ROC) analysis was conducted to compare observer performance under the analog and digital reading conditions. The overall sensitivity of the analog method was 89% versus 78% for the digital method (P < .001), while the specificity values were 96% and 92%, respectively (P > .20). ROC analysis similarly indicated a statistically significant difference in favor of analog radiographs (P = .028). PMID- 8416577 TI - Attenuation properties of lead composite aprons. AB - Traditionally, the absorption properties of protective aprons used in diagnostic radiology have been specified in units of lead equivalent thickness. This is appropriate and accurate when lead is the only high-atomic-numbered component in the apron. In an attempt to manufacture light-weight protective apparel, however, some manufacturers have included other elements with k absorption edges in the energy range of interest, to provide equivalent absorption properties with less weight. With these other high-atomic-numbered elements added, the lead equivalence of the apparel becomes a function of the photon energy. This must be recognized and specified by the supplier, because lead apparel is used in environments other than diagnostic radiology, where the shielding benefits may be substantially less than expected when specifications are based on the diagnostic x-ray energy range. PMID- 8416578 TI - Extracranial head and neck: PET imaging with 2-[F-18]fluoro-2-deoxy-D-glucose and MR imaging correlation. AB - The aim of this study was to define and quantitate the normal anatomy of the extracranial head and neck with 2-[fluorine-18]fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET). This information was used to study 12 patients with primary squamous cell carcinomas. In all cases, the lymphoid tissue of the Waldeyer ring and the palatine and lingual tonsils could be differentiated from the airway, striated muscle, osseous structures, and salivary glands. Striated muscle had markedly less activity than lymphoid or salivary gland tissue. In the 12 patients with primary tumors, FDG PET depicted the tumor as an area of increased activity significantly higher than that of normal tissue. In one instance, FDG PET allowed detection of a tumor not seen at magnetic resonance (MR) imaging or computed tomography. Of the 34 lymph nodes positive for carcinoma, 24 were positive according to MR size criteria and 25 were detected with FDG PET. FDG PET allowed detection of three nonenlarged metastatic nodes that were negative at MR imaging. PMID- 8416579 TI - Cutaneous T-cell lymphoma: CT in evaluation and staging. AB - Among 63 patients with cutaneous T-cell lymphoma (CTCL), 29% (n = 18) had positive computed tomographic (CT) findings, with frequencies of 65% (n = 13) among patients thought to have stages II-IV disease at clinical examination and 12% (n = 5) among patients thought to have stage I. Among eight patients with atypical CTCL variants such as cutaneous large-cell lymphoma, only one had negative findings at CT; extracutaneous disease was not suspected in five before they underwent CT. In contrast, CT findings were positive in only 5% (n = 2) of patients with classic early mycosis fungoides-type CTCL (scaling patches, small epidermotropic CD4+ cells), and CT is unlikely to provide substantial information in this patient subgroup. Contrary to earlier reports, the authors' data suggest that body CT is extremely useful in staging and evaluating patients with CTCL. CT should be included in the evaluation of atypical CTCL variants, Sezary syndrome, advanced-stage mycosis fungoides, and cases in which the CTCL subtype is unclear. PMID- 8416580 TI - Development and evaluation of a needle for percutaneous ethanol injection therapy. AB - To avoid the disadvantages associated with use of the traditional percutaneous transhepatic cholangiography (PTC) needle in ultrasonically guided percutaneous ethanol injection therapy (PEIT) for hepatocellular carcinoma, the authors developed a needle with a three-sided tip and three holes within 3 mm of the tip. In comparison with a PTC needle, the PEIT needle passed into the liver along a straighter line with less resistance and reflected a higher echo from the tip. PMID- 8416581 TI - Book reviews. PMID- 8416582 TI - Relationships between radiology and industry. PMID- 8416583 TI - Air versus barium for monitoring reduction of intussusception. PMID- 8416584 TI - Brain metastases from non-central nervous system tumors: evaluation with PET. AB - Positron emission tomography (PET) with fluorine-18 fluorodeoxyglucose (FDG) was performed in 19 patients with brain metastases from non-central nervous system (CNS) neoplasms and one patient with a primary CNS lymphoma. Various histopathologic types were represented by the primary neoplasms in the patients with metastases. Only 21 of the 31 lesions (68%) were detected with FDG PET as discrete, metabolically active foci (relative to surrounding structures). Six of the nondetected lesions may have been nondiscernible owing to their small size and/or isointensity relative to closely apposed normal gray matter. However, four lesions of at least 1.2 cm in diameter showed frankly decreased FDG accumulation relative to normal brain. These findings suggest that studies of FDG accumulation by a variety of non-CNS neoplasms and their CNS metastases are in order and that extrapolation of the successes of FDG PET in imaging of primary glial tumors to imaging of brain metastases should proceed with caution. PMID- 8416585 TI - Postgraduate radiologic education in Latin America: an international model for combining the resources of organized radiology and a U.S. university. PMID- 8416586 TI - High-resolution (2.6-mm) PET in partial complex epilepsy associated with mesial temporal sclerosis. AB - Eleven patients with medically refractory partial seizures underwent positron emission tomography (PET) with a 600-crystal tomograph and fluorine-18 fluorodeoxyglucose. All patients had been selected for temporal lobe resection, and the side of the epileptogenic focus had been demonstrated with electroencephalography (EEG). Only patients in whom structural lesions had been excluded with magnetic resonance (MR) imaging were studied. Ten of 11 patients were found to have temporal cortical hypometabolism on the same side as the focal abnormality that was demonstrated with EEG. In two patients, PET showed hypometabolism in the mesial temporal cortex only. There were no incorrectly lateralizing PET results. MR imaging showed an abnormality in the corresponding temporal lobe in seven patients. In one patient, both PET and MR images were normal. All patients underwent anterior temporal resection, and histologic examination of resected tissue showed mesial temporal sclerosis in all cases. PMID- 8416587 TI - Functional importance of ventricular enlargement and cortical atrophy in healthy subjects and alcoholics as assessed with PET, MR imaging, and neuropsychologic testing. AB - The authors assessed the relationship between ventricular enlargement, cortical atrophy, regional brain glucose metabolism, and neuropsychologic performance in 10 alcoholics and 10 control subjects. Regional brain glucose metabolism was measured with fluorine-18 fluorodeoxyglucose (FDG) and positron emission tomography (PET). Cortical atrophy and ventricular size were evaluated quantitatively with magnetic resonance (MR) imaging. Alcoholics had decreased brain glucose metabolism and more cortical atrophy but did not have significantly greater ventricular size than did control subjects. The degree of ventricular enlargement and of cortical atrophy was associated with decreased metabolism predominantly in the frontal cortices and subcortical structures in both alcoholics and control subjects. There were no significant correlations between neuropsychologic performance and MR imaging structural changes, whereas various subtest scores were significantly correlated with frontal lobe metabolism. These data show that F-18 FDG PET is a sensitive technique for detecting early functional changes in the brain due to alcohol and/or aging before structural changes can be detected with MR imaging. PMID- 8416588 TI - Unholy alliances: physician investment for self-referral. PMID- 8416589 TI - Self-referral: opportunity for ethical dialogue, appropriate prohibition, and quality assessment. PMID- 8416590 TI - What to do about self-referral? PMID- 8416591 TI - Anti-self-referral legislation: a solution that is long overdue. PMID- 8416592 TI - Endovaginal US and Doppler findings after first-trimester abortion. AB - Endovaginal ultrasound (US) imaging and color Doppler flow imaging techniques were used to evaluate the uterus and its contents and to establish characteristics of a normal post-abortion appearance in 19 women who underwent elective first-trimester abortions. Twenty-two examinations were performed between 2 and 17 days after the procedure. Thirteen of the 22 examinations (59%) showed different amounts of intrauterine material of varying echogenicity. Seven of the 22 examinations (32%) showed a thick endometrial stripe, and only two showed a normal stripe. Color Doppler flow imaging demonstrated typical peritrophoblastic flow in four of eight patients on the second and third days after the abortions were performed. After the third day, flow was observed in only two of 11 patients, and intrauterine material was also seen. These results indicate that intrauterine material and low-impedance flow are frequently observed after an abortion and do not necessarily indicate clinically important retained products of conception. PMID- 8416593 TI - Sedating pediatric patients: is propofol a panacea? PMID- 8416594 TI - Intravenous sedation for MR imaging of the brain and spine in children: pentobarbital versus propofol. AB - The authors present a prospective study of single-agent pediatric sedation regimens for patients older than 2 years of age undergoing magnetic resonance (MR) imaging of the brain and spine. Thirty patients underwent MR imaging after intravenous administration of pentobarbital in successive boluses of 2.5 mg/kg to a maximum of 7.5 mg/kg. Thirty-one patients received an intravenous bolus followed by continuous infusion of propofol. The dosage schedule for propofol was 2 mg/kg (with supplemental 1 mg/kg boluses) followed by continuous infusion of 6 mg/kg per hour. There was no significant difference in the physiologic response to sedation between the two groups, although the magnitude of the drop in pulse was significantly greater in the group receiving propofol. Three patients receiving propofol experienced transient decreases in oxygen saturation, at variable times over the course of the procedure. However, patients recovered significantly faster from sedation with propofol. While propofol may represent a viable alternative to pentobarbital in selected patients, propofol requires constant physician supervision and meticulous technique. PMID- 8416595 TI - Impact of patient-controlled compression on the mammography experience. AB - The authors tested the hypothesis that giving women control over the compression portion of the mammography examination results in a less painful experience, greater overall patient satisfaction, and a radiographic image as good as that produced by means of technologist-controlled compression. One hundred nine women undergoing screening mammography at a hospital-based outpatient clinic were studied. Each underwent two-view, screen-film mammography performed in routine fashion except that, by random assignment, one breast was compressed by the technologist and the other breast, by the patient. Patient-controlled compression was significantly (P = .003) less painful than technologist-controlled compression. Overall patient satisfaction (96% [105 of 109]) and willingness to repeat the experience were extremely high. The majority of images (93.5% [202 of 216]) were rated as having good to excellent compression. With minimal patient education, self-compression resulted in an image at least as good as that produced with technologist-applied compression. Further study of this technique is warranted. PMID- 8416596 TI - 5-HT-1A receptor-mediated effects of antidepressants. AB - 1. Antidepressant (AD) drugs in general induce subsensitivity of behavioural functions associated with activation of 5-HT-1a receptors in animals. 2. Electrophysiological studies in animals in general indicate increased serotonergic transmission after AD administration, mediated partly by increased functioning of post-synaptic 5-HT-1a receptors in the hippocampus. 3. Binding studies have in general shown no change in 5-HT-1a receptor number either pre-or post-synaptically, while results of second messenger studies (inhibition of adenylate cyclase) indicate subsensitivity after AD administration. 4. Human studies also indicate subsensitivity of 5-HT-1a receptors after ADs. PMID- 8416597 TI - Reduced haloperidol/haloperidol ratios after oral haloperidol and decanoate administration in schizophrenics. AB - 1. Haloperidol and reduced haloperidol plasma concentrations were measured in thirteen stable schizophrenic patients that received both oral haloperidol and haloperidol decanoate. 2. Significant correlations between reduced haloperidol/haloperidol ratios from oral haloperidol and haloperidol decanoate occurred at week two and week 16, respectively. 3. The formation of RH was consistent during haloperidol decanoate treatment. PMID- 8416598 TI - Sleep deprivation accelerates the response to nortriptyline. AB - 1. The authors examined the effect of total sleep deprivation (SD) in combination with nortriptyline in 20 patients with major depressive disorder (MDD). Patients underwent a 36-hour SD procedure followed by nortriptyline started on the evening after SD, with ratings for two weeks. 2. Eleven (55%) patients were responders; they showed a rapid and sustained remission after SD, whereas non-responders demonstrated the delayed results expected with nortriptyline. 3. High initial depression scores and absence of depersonalization were associated with response to SD, while being female and middle insomnia were associated with response to the combined regimen. 4. The combination of SD with antidepressants proves to be an effective and safe treatment modality. PMID- 8416599 TI - The sleep EEG and nocturnal hormonal secretion studies on changes during the course of depression and on effects of CNS-active drugs. AB - 1. The sleep EEG and nocturnal hormone secretion were studied simultaneously in normal male controls and in male patients with major endogenous depression before treatment with tricyclics and after recovery and drug cessation. 2. Several studies were performed in normal male controls to investigate the effect of antidepressants (brofaromine, moclobemide, amitriptyline, clomipramine and trimipramine) and of neuropeptides (CRH and the ACTH (4-9) fragment analog ebiratide) on the sleep EEG and sleep-associated hormone secretion. 3. Elevated cortisol and blunted testosterone secretion are state markers of acute depression, whereas sleep EEG, GH and prolactin variables do not show marked differences between acute depression and recovery. Except for trimipramine, all antidepressants investigated suppress REM sleep. No systematic relationship between the sleep EEG and endocrine effects of antidepressants is detectable. Pulsatile application of CRH in controls mimicks some of the neurobiological symptoms of acute depression. More shallow sleep occurs under ebiratide, whereas hormonal secretion remains unchanged. 4. Our data demonstrate that antidepressants exert distinct effects on sleep. However, these substances do not induce changes in sleep structure which persist after their withdrawal in remitted patients. Pulsatile application of neuropeptides leads to specific effects on CNS activity which are not mediated by changes of peripheral hormone secretion. The view that CRH plays a key role in the pathophysiology of affective disorders is corroborated. PMID- 8416600 TI - Iron, melanin and dopamine interaction: relevance to Parkinson's disease. AB - 1. Interaction between iron and melanin may provide a reasonable explanation for the vulnerability of the melanin containing dopaminergic neurons in the substantia nigra (SN) to neurodegeneration in Parkinson's disease (PD). 2. Scatchard analysis of the binding of iron to synthetic dopamine melanin revealed a high-affinity (KD = 13 nM) and a lower affinity (KD = 200 nM) binding sites. 3. The binding of iron to melanin is dependent on the concentration of melanin and on pH. 4. Iron chelators, U74500A, desferrioxamine and to a lesser extent 1,10 phenanthroline and chlorpromazine could displace iron from melanin. In contrast, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its metabolite 1-methyl-4 phenyl-pyridinium (MPP+), which cause Parkinsonism, were unable to displace iron. 5. Melanin alone reduced lipid peroxidation in rat cortical membrane preparations. However, iron induced lipid peroxidation, which could be inhibited by desferrioxamine, was potentiated by melanin. 6. Iron bound to neuromelanin in melanized dopamine neurons was detected only in parkinsonian brains and not in controls. The interaction of iron with neuromelanin as identified by x-ray defraction technique was identical to iron interaction with synthetic dopamine melanin. 7. In the absence of an identified exogenous or endogenous neurotoxin in idiopathic Parkinson's disease, iron-melanin interaction in the SN may serve as a candidate for the oxygen-radical induced neurodegeneration of the melanin containing dopaminergic neurons. PMID- 8416601 TI - Effects of single and repeated immobilization stress on corticotropin-releasing factor concentrations in discrete rat brain regions. AB - 1. Corticotropin-releasing factor (CRF) has been suggested to regulate many responses to stress. The authors investigated the effect of single and repeated stress on brain CRF immunoreactivity (CRF-ir) and plasma corticosterone levels in rats, using radioimmunoassay. 2. Single immobilization stress significantly increased plasma corticosterone levels but did not change CRF-ir in the discrete brain regions at all. Repeated immobilization stress (a 180 min session, once a day for 10 days) did not affect plasma corticosterone levels at 24 hr poststress. However, it increased CRF-ir in the median eminence (ME) though not in the other brain regions. 3. The increased level of CRF in the ME after chronic intermittent stress suggests that repeated stimulation by stress may increase the storage pool of CRF in the ME. PMID- 8416602 TI - Mechanisms of drug actions against neuronal damage caused by ischemia--an overview. AB - 1. Oxygen and energy deficits induces a cascade of pathological processes which lead to neuronal dysfunction and cell death. 2. The pathogenesis of ischemia induced neuronal damage includes a disturbed calcium homeostasis, an excessive release of EAA and an enhanced formation of free oxygen radicals. 3. Calcium antagonists inhibit Ca2+ influx into the neuronal cell via VSCC. 4. Glutamate antagonists reduce intracellular Ca2+ concentration by inactivation of NMDA receptor-associated calcium channels (NMDA antagonists) or AMPA/quisqualate receptor-linked sodium channels (non-NMDA antagonists). 5. Furthermore, oxygen radical scavengers can avoid neuronal damage. 6. Agonists of the adenosinergic and serotonergic transmitter systems contribute to neuroprotection by hyperpolarization of the neuronal membrane due to an increase of K+ permeability. PMID- 8416603 TI - 5-HT1A receptor ligands in animal models of anxiety, impulsivity and depression: multiple mechanisms of action? AB - 1. Preclinical and clinical studies suggest that 5-HT1A receptor agonists are a new class of mixed anxiolytics/antidepressants with, possibly, impulsivity reducing properties. 2. The anxiolytic effects of 5-HT1A receptor agonists result predominantly from an interaction with presynaptic 5-HT1A receptors (resulting in a decrease of serotonergic transmission), whereas the antidepressive and, possibly, the impulse control enhancing effects, result predominantly from an interaction with postsynaptic 5-HT1A receptors. 3. These proposed mechanism(s) of action fit well with the generally held view that anxiety is the result of a hypersensitive 5-HT system; whereas impulsivity and depression is the result of a hyposensitive 5-HT system. 4. However, it appears very likely that activation of pre- and postsynaptic 5-HT1A receptors is additionally involved in the antidepressive and impulse control enhancing effects, on the one hand, and in the anxiolytic effects of these compounds, on the other hand. 5. These latter, seemingly paradoxical, findings can be explained by assuming that (1) the presynaptic mechanism reflects an anxiolytic component in the animal models of impulsivity and depression, (2) antagonism of postsynaptic 5-HT1A receptors by these compounds contributes to their anxiolytic effects, (3) postsynaptic 5-HT1A and 5-HT2 receptors have functionally opposing effects or, alternatively, that (4) downregulation of postsynaptic 5-HT2 receptors contributes to the therapeutic effects of these compounds. PMID- 8416604 TI - Ethics in current medical imaging. PMID- 8416605 TI - Sonography of the shoulder in patients with tears of the rotator cuff: accuracy and value for selecting surgical options. AB - OBJECTIVE: The management of patients with signs and symptoms referable to the rotator cuff depends on the presence of cuff injury and the size of the tear. Treatment options include conservative nonsurgical management for patients with an intact or partially torn cuff, arthroscopic decompression of the coracoacromial space for those not responding to nonsurgical management, and a range of surgical techniques to repair full-thickness tears. This study was designed to determine whether sonographic evaluation with classification of the extent of cuff injury is accurate for purposes of treatment planning. SUBJECTS AND METHODS: Preoperative sonography of the rotator cuff was performed on 225 patients, and findings were classified into intact, partial tear, small full thickness tear, large full-thickness tear, and massive tear groups. Surgical correlation with the predicted sonographic classification was provided by arthroscopic inspection or open surgery. RESULTS: The sonographic findings were surgically confirmed for 206 (92%) of the 225 patients. More extensive cuff injury was encountered during surgery than had been predicted sonographically in 11 patients (5%); less extensive injury than predicted was found during surgery in eight patients (4%). CONCLUSION: Our results show a high correlation between the sonographic classification of rotator cuff injury and the surgical findings. The selection of appropriate treatment programs can be reliably based on the sonographic classification. PMID- 8416606 TI - Stress fractures of the tarsal navicular bone: CT findings in 55 cases. AB - OBJECTIVE: The purpose of this article is to present the CT findings in 55 cases of tarsal navicular stress fracture before and after treatment and to describe the CT protocol used. MATERIALS AND METHODS: Fifty-five navicular stress fractures in 54 patients whose initial and follow-up CT scans were available for study were retrospectively reviewed. In most cases, contiguous 1.5-mm axial and 3 mm coronal scans of the navicular bone had been obtained. Both feet were scanned for comparison. Follow-up scans were obtained between 4 weeks and 6 years. RESULTS: On review, fractures were evident in all cases, but six small fractures (11%) were missed at the initial interpretation. All fractures involved the central third of the proximal dorsal margin of the navicular bone. Fifty-three fractures (96%) were partial. Forty-three partial fractures were linear, five were linear with bone fragments, and five were rim defects with ossicles. In 13 cases (24%) the fracture was small, 10% or less of bone height. The earliest sign of healing, slight dorsal cortical bridging, was seen in three of eight cases in which follow-up was done at 6 weeks. Firm cortical union was noted in 10 (32%) of 31 by 4 months. Nonunion occurred in 12 and was indicated by the persistence of the fracture gap and lack of cortical healing. Medullary cysts (five) and cortical notching (two) were noted to persist after complete healing. CONCLUSION: CT scanning is a suitable method for detecting navicular stress fracture and for performing follow-up examinations. Small fractures may be overlooked owing to lack of familiarity with their appearance. PMID- 8416607 TI - Intrinsic and extrinsic carpal ligaments: evaluation by three-dimensional Fourier transform MR imaging. AB - OBJECTIVE: As evaluation of the anatomic and functional integrity of intrinsic and extrinsic carpal ligaments with conventional imaging methods is difficult, we designed a study to evaluate the ability of three-dimensional Fourier transform MR imaging to show the carpal ligaments. SUBJECTS AND METHODS: We obtained coronal MR images of 15 cadaveric wrists and 15 wrists of patients, using three dimensional volume acquisition with a gradient-recalled echo sequence and a 1.5-T magnet. The MR findings were compared with the findings on dissection in the 15 cadaveric wrists and with the surgical findings in eight patients. RESULTS: All the volar ligaments had a striated appearance on MR images, with alternating bands of low and intermediate signal intensity. No tears of the extrinsic ligaments were seen. The intrinsic scapholunate and lunotriquetral ligaments were seen as structures with more homogeneously intermediate intensity attaching to their adjacent bone through high-signal-intensity hyaline cartilage. For the cadaveric wrists, MR indications of a tear of the scapholunate ligament were true positive in three and false-negative in three; indications of a tear of the lunotriquetral ligament were true-positive in five, false-positive in two, and false-negative in one. For the eight patients with surgical confirmation, the MR findings regarding tears were true-positive in two and true-negative in six. Neither surgery nor MR imaging showed any tears of the scapholunate ligament. CONCLUSION: Three-dimensional Fourier transform MR imaging with thin slices provides the resolution necessary to visualize the anatomic detail of the extrinsic and intrinsic ligaments of the wrist, but additional clinical experience with this technique will be required to determine its diagnostic capabilities. PMID- 8416608 TI - Viral infections of the CNS in children: imaging features. AB - In this paper, we review the imaging findings associated with the major viral infections of the CNS in infants and children. We approach these infections by grouping them into several categories: congenital infections in neonates, aseptic meningitis and encephalitis, acute disseminated encephalomyelitis and other postviral syndromes in older children, and HIV infection. PMID- 8416609 TI - Posttraumatic carotid artery dissection in children: evaluation with MR angiography. PMID- 8416610 TI - An unusual first-trimester sonographic finding associated with development of hydatidiform mole: the hyperechoic ovoid mass. PMID- 8416611 TI - Aneurysm of a nonpatent ductus arteriosus in a neonate: CT findings. PMID- 8416612 TI - Pseudotumor cerebri: CT findings and correlation with vision loss. AB - OBJECTIVE: The purpose of this study was to determine if orbital and cerebral CT can be used to distinguish normal patients from those with pseudotumor cerebri, and to correlate CT findings with the severity of visual impairment. SUBJECTS AND METHODS: Seventeen patients with a clinical diagnosis of pseudotumor cerebri were compared with 20 age- and sex-matched control subjects. Thin-section coronal and axial CT scans of the orbit and whole-brain axial CT scans were available for all subjects. The diameter of the optic nerve sheath, the degree of reversal of the optic nerve head, the presence and degree of empty sellae, and ventricular and sulcal sizes were evaluated without knowledge of whether or not the subject had pseudotumor cerebri. The same parameters were compared for two subgroups of patients with pseudotumor cerebri: those with mild vision loss and those with severe vision loss. RESULTS: Patients with pseudotumor cerebri had significantly larger optic nerve sheaths than did control subjects (6.5 +/- 0.83 mm vs 5.4 +/- 0.69 mm). Radiologic evidence of papilledema with reversal of the optic nerve head was found in 12 of 17 patients compared with one of 20 control subjects. An empty sella was seen more frequently and to a greater degree in patients with pseudotumor cerebri than in control subjects (16 vs seven). Eight of nine patients with severe vision loss and four of eight patients with mild to moderate vision loss had reversal of the optic nerve head; the degree was greater in the group with severe vision loss. No difference in ventricular size or sulcal enlargement was seen between any of the groups. The opening CSF pressures of the two groups with vision loss were not significantly different (348 +/- 80 mm H2O vs 391 +/- 98 mm H2O). CONCLUSION: In addition to the role of CT in excluding intracranial disease as a basis for the clinical syndrome of pseudotumor cerebri, thin-section CT of the orbits reveals a constellation of findings, including enlarged optic nerve sheaths, reversal of the optic nerve head, and empty sellae in patients with pseudotumor cerebri. Furthermore, severe vision loss in these patients correlates with more frequent and more severe reversal of the optic nerve head. PMID- 8416613 TI - Late CT findings in brain trauma: relationship to cognitive and behavioral sequelae and to vocational outcome. AB - OBJECTIVE: The purpose of this study was to determine the degree of correlation between cerebral atrophy observed on CT scans after severe blunt brain trauma and later neuropsychological status, as well as to evaluate the relative prognostic values of a number of indexes of cerebral atrophy. MATERIALS AND METHODS: The study group comprised 32 previously healthy men 18-65 years old who had severe blunt trauma of the brain (initial scores on Glasgow coma scale of 7 or less). Their cognitive and behavioral statuses were evaluated when the patients were discharged from the hospital, which occurred when the recovery process showed a plateau. Overall vocational status was evaluated 1 year after discharge. The clinical evaluation was performed by a multidisciplinary team. Multiple linear indexes derived from brain CT scans obtained about 3 months after injury in patients with blunt brain trauma were correlated with cognitive and behavioral sequelae of brain damage and with vocational placement, as evaluated by a rehabilitation team about 1 year after trauma. RESULTS: A high correlation was found between the width of the third ventricle and outcome. The prognostic value of the width of the third ventricle was superior to that of any other index studied, and it correlated best with late cognitive status (Spearman r = .57, p < .01). CONCLUSION: The width of the third ventricle is a useful prognostic index in cases of diffuse brain trauma. It indicates diencephalic atrophy, caused either by diffuse axonal injury or by hypoxia. It may indicate a role of diencephalic structures in higher cortical functions. PMID- 8416614 TI - Disseminated histoplasmosis in AIDS: findings on chest radiographs. AB - OBJECTIVE: Our objective was to determine the findings of disseminated histoplasmosis on chest radiographs of patients with AIDS. MATERIALS AND METHODS: Chest radiographs of 50 AIDS patients with documented extrapulmonary histoplasmosis were analyzed retrospectively. The radiographs were evaluated for the presence of parenchymal opacities, pleural effusions, adenopathy, cavitation, and calcified granulomas and lymph nodes. A modification of the International Labour Office scheme was used to classify parenchymal abnormalities as nodular, linear or irregular, reticulonodular, or air-space opacities. RESULTS: Abnormalities were present on radiographs in 23 patients. Nodular opacities were present in 10 patients and were diffusely distributed in nine patients. Linear or irregular opacities were present in seven patients, with diffuse distribution in four and limited involvement in three. Air-space opacities were present in seven patients; the distribution varied from segmental to diffuse involvement of the lung. Small pleural effusions were present in five patients. Adenopathy and Kerley's B lines were each present in three patients. In 27 patients, the chest radiographs were normal. Four of these patients had clinical or microbiological evidence of lung involvement. CONCLUSION: The chest radiographic findings of disseminated histoplasmosis in AIDS patients are varied and nonspecific. The presence of diffuse nodular or linear/irregular opacities in an AIDS patient, especially one who resides in or has resided in an endemic area, should suggest the possibility of disseminated histoplasmosis. Normal findings on chest radiographs do not exclude disseminated infection or lung involvement. PMID- 8416615 TI - MR imaging of the brain in patients with AIDS: value of routine use of i.v. gadopentetate dimeglumine. AB - OBJECTIVE: A prospective study was conducted to explore the value of routine administration of IV gadopentetate dimeglumine for MR imaging of the brain in patients with AIDS. SUBJECTS AND METHODS: Over a 19-month period, MR images of the brain in 51 consecutive AIDS patients were obtained routinely both with and without IV gadopentetate dimeglumine. Unenhanced and contrast-enhanced images from the resulting 63 studies were viewed together. Findings were classified into one or more of three categories: normal, mass or focal lesions, or white matter disease. The number of focal or mass lesions was recorded. Lesion conspicuity on the unenhanced and enhanced images was compared. Ventricular enlargement was also graded. Available medical records and laboratory data of the patients were reviewed. RESULTS: Of the 63 MR studies reviewed, 39 (62%) were abnormal. In no case was a normal unenhanced MR study rendered abnormal after the administration of gadopentetate dimeglumine. Overall, T2-weighted images showed twice as many focal or mass lesions than contrast-enhanced T1-weighted images did. Most lesions detected on the T2-weighted images did not show enhancement with contrast material. White matter disease was the most common abnormality detected. The group of patients with white matter disease also had the highest occurrence of ventriculomegaly. CONCLUSION: Our study does not support the routine use of gadopentetate dimeglumine for MR imaging of the brain in patients with AIDS. Our experience emphasizes the importance of a normal T2-weighted image. PMID- 8416616 TI - More nominal dysphasia in medical nomenclature. PMID- 8416617 TI - Neurofibromatosis: MR imaging findings involving the head and spine. AB - MR imaging can be used to identify abnormalities of the head and spine in patients with neurofibromatosis. In this pictorial essay, we illustrate the craniospinal MR imaging findings in a large series of patients with neurofibromatosis. PMID- 8416618 TI - Fluid flow during percutaneous drainage procedures: an in vitro study of the effects of fluid viscosity, catheter size, and adjunctive urokinase. AB - OBJECTIVE: An in vitro study was performed to determine the range of flow times of different bodily fluids through catheters of different diameters and to test the hypothesis that urokinase might decrease the viscosity of purulent material. MATERIALS AND METHODS: A standard viscometer was used to measure the viscosities of water, blood, pseudocyst fluid, purulent material, and purulent material with admixed urokinase. For each fluid, Poiseuille's law was used to calculate the kinematic viscosity, from which theoretical drainage times through seven different sizes of catheters were calculated. These theoretical times were compared with the actual measured values to verify that flow was according to Poiseuille's law. RESULTS: The calculated kinematic viscosities (in 10(-6) stokes) were as follows: water, 0.695 +/- 0.006; pseudocyst fluid, 2.185 +/- 0.008; blood, 3.001 +/- 0.049; abscess fluid without urokinase, 5.729 +/- 0.064; and abscess fluid with urokinase, 4.416 +/- 0.070. The viscosity of abscess fluid decreased by 23% with the addition of urokinase. Drainage time was considerably shorter with larger catheters. CONCLUSION: Flow of various bodily fluids, including pus, is according to Poiseuille's law, confirming that for more viscous fluid, larger catheters provide more rapid drainage. Urokinase decreases viscosity of purulent material and increases flow for all sizes of catheters. PMID- 8416619 TI - Safety of intracavitary urokinase with percutaneous abscess drainage. AB - OBJECTIVE: Percutaneous drainage of abscesses is an effective treatment, but the success rate is lower for abscesses that have septa and are multilocular. Several clinical and in vitro studies suggest urokinase may be useful in such cases. Our study was designed to determine the safety of urokinase administered into an abscess cavity during the course of percutaneous drainage. SUBJECTS AND METHODS: Our study included 26 consecutive patients with 31 abscesses treated with percutaneous drainage. Exclusion criteria included age less than 18 or more than 95 years, CNS disorders (e.g., tumor, vascular problems), coagulation impairments, hepatic failure, pregnancy, and abscesses in the spleen, pancreas, or interloop area. Three doses were used: group 1 (nine patients), 1000 IU of urokinase per centimeter of abscess diameter; group 2 (11 patients), 2500 IU of urokinase per centimeter of abscess diameter; and group 3 (nine patients), 5000 IU of urokinase per centimeter of abscess diameter. These doses were administered every 8 hr for 3 days along with percutaneous drainage. Charts were reviewed to determine success and to detect adverse clinical events. Studies included sequential CT scans; serial serum determinations of hematocrit, prothrombin time, partial thromboplastin time, platelet count, fibrinogen levels, and levels of fibrin degradation products; and serial laboratory analysis of purulent material for fibrinogen and fibrin degradation products. Percutaneous drainage was considered successful if no surgical intervention was required. RESULTS: Our results showed no significant change in hematologic studies and no bleeding complications. Analysis of purulent material indicated that urokinase remained active in the abscess milieu. Drainage was successful in seven of 11 patients in group 1, all nine patients in group 2, and 10 of 11 patients in group 3. All eight abscesses with septa were successfully drained. CONCLUSION: Intracavitary urokinase can be given safely during percutaneous drainage of an abscess, with no associated bleeding complications or changes in coagulation parameters. PMID- 8416620 TI - Value of having a cytopathologist present during percutaneous fine-needle aspiration biopsy of lung: report of 55 cancer patients and metaanalysis of the literature. AB - OBJECTIVE: Percutaneous fine-needle aspiration biopsy is an accepted procedure for diagnosing intrathoracic malignant disease. The value of having a cytopathologist present during the procedure was studied with respect to the number of needle passes, the accuracy of the procedure, and complications. A metaanalysis was performed on the combined results of the present and previous series. SUBJECTS AND METHODS: We analyzed data from 55 adult patients who underwent percutaneous CT-guided fine-needle (22-gauge) aspiration biopsy of the lung for a lesion that either the biopsy or another subsequent invasive procedure showed to be nonlymphomatous and malignant. A cytopathologist was present for the first 25 procedures and not present for the next 30 procedures. When present, the cytopathologist stained the aspirated material with toluidine blue and gave an immediate opinion on the diagnostic adequacy of the specimen based on microscopic evaluation. If appropriate, the radiologist obtained additional biopsy specimens. When a cytopathologist was not present, the radiologist assessed the adequacy of the specimen by gross examination. A metaanalysis was performed of 211 cases from the present study and two previous series with respect to the effect of the presence of a cytopathologist on the diagnostic accuracy of the procedure. RESULTS: Biopsy specimens showed cancer in 25 (100%) of 25 patients when obtained while the cytopathologist was present, and in 24 (80%) of 30 patients when obtained while the cytopathologist was absent (p < .05, Fisher exact test). No significant differences in the number of needle passes performed or in the frequency of pneumothorax when aerated lung was traversed were noted between the two groups. Although two previous studies showed nonsignificant trends toward increased accuracy of thoracic fine-needle aspiration when a cytopathologist participated in the procedure, metaanalysis revealed significantly increased accuracy when a cytopathologist was present (p < .02, Mantel-Haenszel chi 2 test). CONCLUSION: An accurate diagnosis from fine-needle aspiration biopsy of intrathoracic cancer was more likely when a cytopathologist was present than when not present during the procedure. PMID- 8416621 TI - Devices for transcatheter closure of intracardiac defects. AB - Transcatheter closure of intracardiac defects is an investigational procedure that is in use at a number of centers in North America and Europe. A radiologist should be able to recognize these devices on a chest radiograph, understand their actual physical appearance, and be able to recognize their expected location. This essay summarizes the indications for, technical aspects of, and radiologic appearance of these devices. PMID- 8416622 TI - An automated radiology reporting system that uses HyperCard. AB - SCRIBE is an automated radiology reporting system that uses the HyperCard environment on Macintosh computers. Radiologic findings and anatomic terms are presented in graphic form, and the appropriate terms are selected by using a trackball or touch-sensitive video screen. Additional lists of more specific terms and differential diagnoses can be requested by the user for abnormal findings. The system is suited to the reporting of plain films and is being used in the emergency room of a large academic radiology department. Advantages of the system include low cost, operational familiarity to Macintosh users, and elimination of transcription costs. Finished reports are immediately available in both printed and electronic forms. PMID- 8416623 TI - Early history of diagnostic ultrasound: the role of American radiologists. PMID- 8416624 TI - Extravasation injury with nonionic contrast material. PMID- 8416625 TI - High-resolution sonography of the diaphragmatic pleura. PMID- 8416626 TI - Benign fibrous mesothelioma of the pleura: MR findings. PMID- 8416627 TI - Bronchiolitis obliterans organizing pneumonia with migratory infiltrates: a late complication of radiation therapy. PMID- 8416628 TI - Conversion of gastrostomy tube to gastrojejunostomy tube by using a peel-away sheath. PMID- 8416629 TI - Bacillary angiomatosis in a patient with HIV infection. PMID- 8416630 TI - Dilated small bowel simulating a submucosal gastric mass. PMID- 8416631 TI - Use of transperineal sonography to locate an obstructing urethral calculus. PMID- 8416632 TI - Blood-fluid level within a renal cyst. PMID- 8416633 TI - Adrenal pheochromocytoma. PMID- 8416634 TI - Diagnostic success of bronchoscopic biopsy in immunocompromised patients with acute pulmonary disease: predictive value of disease distribution as shown on CT. AB - OBJECTIVE: The purpose of the study was to determine if the distribution of pulmonary opacities on CT scans could be used to predict the outcome of bronchoscopic biopsy procedures in immunocompromised non-AIDS patients with acute pulmonary complications. MATERIALS AND METHODS: Thirty-three consecutive immunocompromised patients without AIDS who had acute pulmonary complications and who had had CT, bronchoscopic biopsy procedures, and proved diagnoses were included in the study. The distribution and dominant pattern of pulmonary opacities on CT were assessed independently by two observers. The pathologic diagnoses were invasive aspergillosis (eight), Candida pneumonia (six), bronchiolitis obliterans with or without organizing pneumonia (six), drug-induced lung disease (four), Pneumocystis carinii pneumonia (four), cytomegalovirus pneumonia (three), pulmonary hemorrhage (one), and recurrent lymphoma (one). RESULTS: The results of bronchoscopic techniques established a specific diagnosis in 17 patients (52%). In the remaining 16 patients, results of bronchoscopic biopsy could not be used to establish a specific diagnosis; open lung biopsy (15 patients) or transthoracic needle biopsy (one patient) were required for diagnosis. The results of bronchoscopic procedures were diagnostic more often in patients in whom pulmonary opacities involved the central third of the lung than in patients in whom the central third was spared (70% vs 23%, p = .02). Results were diagnostic more often in cases in which the causes of acute pulmonary complications were infectious than in cases in which the causes were noninfectious (71% vs 17%, p < .005). CONCLUSION: We conclude that the presence or absence of central disease as shown by CT can be used to suggest whether results of bronchoscopic procedures in immunocompromised non-AIDS patients will be diagnostic. PMID- 8416635 TI - Subtle clue to duplication of inferior vena cava. PMID- 8416636 TI - CT-guided excision of osteoid osteoma. PMID- 8416637 TI - In utero clue to congenital lethal osteogenesis imperfecta. PMID- 8416638 TI - Left paracardiac mass caused by dilated pericardiacophrenic vein: report of four cases. AB - OBJECTIVE: The pericardiacophrenic vein is a possible route of collateral circulation when either the superior or the inferior vena cava is obstructed. When the vein is dilated, it can cause an abnormal appearance on chest radiographs. We describe four such patients. MATERIALS AND METHODS: We studied four patients who had abnormalities on conventional chest radiographs due to a grossly dilated pericardiacophrenic vein, confirmed by means of venacavography, CT, or MR imaging. We analyzed the cause of the dilatation and the radiologic findings associated with the dilated pericardiacophrenic vein. RESULTS: The cause of the dilated pericardiacophrenic vein was the membranous obstruction of the inferior vena cava in all four patients. Chest radiographs revealed an undulating vascular shadow (in two cases) or a shadow of several masses (in one case) along the left border of the heart or a poorly defined haziness mimicking a pulmonary parenchymal infiltrate (in one case). A direct communication between the left hepatic vein and the left inferior phrenic vein was seen near their insertion into the inferior vena cava in three cases. Hepatic venous outflow and some of the systemic venous return were directed to this left inferior phrenic pericardiacophrenic route. CONCLUSION: When obstruction of the inferior vena cava is clinically suspected and the chest radiograph shows abnormalities at the left paracardiac area, a dilated pericardiacophrenic vein due to obstruction in the suprahepatic portion of the inferior vena cava, most likely a membrane, should be considered in the differential diagnosis. PMID- 8416639 TI - Value of fluoroscopy in patients with suspected bilateral hemidiaphragmatic paralysis. PMID- 8416640 TI - Blistered Feet. PMID- 8416641 TI - Subclavian vein thrombosis detected with spiral CT and three-dimensional reconstruction. PMID- 8416642 TI - MR angiography of the portal and hepatic venous systems: preliminary experience with echoplanar imaging. AB - OBJECTIVE: The purpose of this study was to evaluate the ability of echoplanar MR angiography to depict the major hepatic and portal venous structures. SUBJECTS AND METHODS: Echoplanar and conventional MR angiographic examinations were performed in 10 subjects (seven healthy volunteers, three patients with focal hepatic lesions). A gradient-recalled echo (GRE) time-of-flight technique (125/10 [TR/TE], 90 degrees flip angle) was used for echoplanar angiography. Eight complete single-excitation images were acquired at each level in 1.5 sec and then collapsed into a single maximal intensity projection. Conventional time-of-flight MR angiography (34/13, 30 degrees flip angle) also was performed. The vascular anatomy from the right atrium to the splenic vein was imaged (6-mm contiguous levels) in three 10.5-sec breath-holds with echoplanar imaging, as compared with seven 11.5-sec breath-holds with conventional MR angiography. Echoplanar and conventional images were compared quantitatively and qualitatively. RESULTS: Echoplanar imaging was 61% faster than conventional MR angiography. Vessel-to liver signal-intensity ratios were significantly higher for echoplanar imaging (p < .0001), signal-to-noise ratios were significantly higher for conventional MR angiography (p < .0001), and contrast-to-noise ratios were comparable. Qualitatively, echoplanar imaging and conventional MR angiography provided similar anatomic information about the hepatic and portal veins. CONCLUSION: Angiograms of the hepatic and portal venous systems that are of diagnostic quality can be acquired much more quickly with echoplanar imaging than with conventional MR angiography. PMID- 8416643 TI - Recurrent bleeding after variceal hemorrhage: predictive value of portal venous duplex sonography. AB - OBJECTIVE: Risk assessment of recurrent variceal bleeding is essential for therapeutic decisions and is usually performed by endoscopy of the upper gastrointestinal tract. We studied the value of portal venous duplex sonography in predicting subsequent variceal bleeding in patients with cirrhosis. SUBJECTS AND METHODS: Thirty patients with cirrhosis who received sclerotherapy because of acute variceal hemorrhage for the first time (hemorrhage group), 30 patients with cirrhosis who had no previous hemorrhage (nonhemorrhage group), and 30 control subjects were examined prospectively. With the use of portal duplex and color Doppler sonography, flow direction, flow velocity, vein diameter, and response to respiration of portal vein vessels were measured and portosystemic collaterals and thrombosis of portal vessels were visualized. The results of these measurements and imaging findings were combined into a Doppler sonoscore. At entry into the study, all patients were classified on the basis of a sonoscore as having a low (sonoscore, < 4) or a high (sonoscore, > or = 4) risk for subsequent hemorrhage. During a mean follow-up period of 2 years (range, 15-36 months), the predictive value of this Doppler sonoscore was studied. RESULTS: In the hemorrhage group, the prevalence of recurrent hemorrhage was 40%, despite sclerotherapy, and the mortality rate was 60%. In patients with a Doppler sonoscore of 4 or more, the prevalence of recurrent hemorrhage was 67%, whereas in patients with a score less than 4, the prevalence was only 22% (p < .02). After sclerotherapy, endoscopic criteria showed no significant correlation with the prevalence of bleeding. In the nonhemorrhage group, the prevalence of variceal hemorrhage occurring was 13%, and Doppler sonographic criteria showed no significant correlation with the prevalence of subsequent hemorrhage. CONCLUSION: We conclude that Doppler sonography, performed after the first occurrence of variceal hemorrhage, provides useful prognostic information regarding the risk of recurrent hemorrhage. If these results are confirmed, Doppler sonography may be used to select the best method of treatment. PMID- 8416644 TI - Metastases to the pancreas and peripancreatic lymph nodes from carcinoma of the right side of the colon: CT findings in 12 patients. AB - OBJECTIVE: Our objective was to describe the CT findings of metastases to the pancreas from carcinoma of the colon and to discuss the pathways of metastasis based on the anatomic relationship between the mesocolon and the pancreas. MATERIALS AND METHODS: Clinical features and CT scans of 12 patients who had proved metastases to the pancreas from adenocarcinoma of the colon were retrospectively reviewed to define the characteristics of pancreatic lesions. The primary tumors were in the cecum (three patients), ascending colon (five patients), and transverse colon (four patients). Direct extension of the tumor to the pancreas was excluded. Metastases were diagnosed by aspiration or surgical biopsy. RESULTS: Seven patients (58%) had obstruction of the bile duct and/or pancreatic duct. Four others had symptoms related to the mass, including pain and gastrointestinal obstruction. In eight patients (67%), metastatic tumors involved the pancreas as a focal mass; in the other four (33%), the masses were lobulated and engulfed the pancreas and were indistinguishable from peripancreatic nodal disease. The masses were hypodense in nine patients (75%) and isodense in three patients (25%). Extra-pancreatic metastatic disease was seen in nine patients (75%). CONCLUSION: Clinical features and CT findings in patients with pancreatic metastases from carcinoma of the colon are similar to those of primary pancreatic ductal adenocarcinomas. The diagnosis of metastasis should be considered when a patient has a pancreatic mass and a history of colon carcinoma. PMID- 8416645 TI - Edward B. D. Neuhauser Lecture. Pediatric neuroradiology: its evolution as a subspecialty. AB - Pediatric neuroradiology emerged as the first formal subspecialty of pediatric radiology during the late 1960s. The history of its development as an unusual and effective combination of an age-related and a specific organ-directed clinical subspecialty, and the considerable technical adaptation and innovation within the diagnostic imaging so required, merits its inclusion within the history of the modern matrix of radiology. This Neuhauser Lecture outlines the odyssey of this subspecialty until the present: the adaptation of techniques and equipment to accommodate imaging of patients of all sizes--from 1-kg infants to young adults- and the understanding of the wide spectrum of CNS diseases, many often extremely complex. The evolution of this special body of knowledge and experience, the established and productive fellowship programs, and the significant part pediatric neuroradiology now plays in major scientific and professional associations and societies have led to this subspecialty becoming a lifelong persuasion for a growing number of radiologists. PMID- 8416646 TI - CT of blunt trauma to the bowel and mesentery. AB - The high mortality and morbidity rates associated with traumatic rupture of the hollow viscera have been attributed to the clinical difficulty in establishing an early diagnosis. CT has been shown to be accurate for detecting bowel and mesenteric injuries caused by blunt trauma and may be useful in predicting the need for either surgical repair or conservative management. However, many major gastrointestinal injuries have subtle CT findings. In this pictorial essay, we illustrate the broad spectrum of gastrointestinal abnormalities that can be shown by CT in patients with blunt abdominal trauma. PMID- 8416647 TI - Gastric syphilis: radiologic findings. AB - Syphilis, a venereal infection caused by the spirochetal bacterium Treponema pallidum, has long been considered a primary public health concern in the United States. With the onset of the antibiotic era, the prevalence of the disease dramatically plummeted, as did interest in its radiologic manifestations. Rolfs and Nakashima [1] have shown that the prevalence of primary and secondary syphilis increased 34% from 1981 to 1989, to its highest level since 1949. Given this dramatic increase, classic manifestations of syphilis may warrant renewed attention. In its secondary and tertiary stages, syphilis can cause a wide range of gastric lesions that can mimic many other entities, from gastritis or benign ulcer disease to gastric carcinoma. Indeed, the acute gastritis of early secondary syphilis produces the earliest radiologically detectable signs of the disease. Cases of gastric syphilis submitted to the Armed Forces Institute of Pathology and cases drawn from the University of Cincinnati teaching file are used to illustrate the varied findings in this disease. PMID- 8416648 TI - Cathode rays and controversy. PMID- 8416649 TI - Leukemic infiltration of the gallbladder wall mimicking acute cholecystitis. PMID- 8416650 TI - Diagnosis of arteriogenic impotence: efficacy of duplex sonography as a screening tool. AB - OBJECTIVE: The results of duplex sonography of the cavernosal artery were compared with the results of pharmacoarteriography in a series of impotent men in order to assess the validity of sonography as a screening tool for the diagnosis of arteriogenic impotence. SUBJECTS AND METHODS: Duplex sonography was performed in 30 impotent men after intracavernosal injection of papaverine and phentolamine. Maximal acceleration, peak systolic velocity, and resistive index were determined for the cavernosal artery. All patients had selective pharmacoarteriography and cavernosometry, which were used to diagnose arterial disease (bilateral hemodynamically significant stenoses or occlusions), suspected arteriolar dysfunction (inconspicuous helicine arterioles), or venoocclusive insufficiency (pharmacologic maintenance erectile flow greater than 25 ml/min). Seventeen patients had arterial insufficiency, four had arteriolar dysfunction, and nine had no significant arterial disease. RESULTS: Significant differences in Doppler measurements were found between patients without significant arterial disease and those with arterial insufficiency, including peak systolic velocity (50 +/- 25 cm/sec vs 33 +/- 17 cm/sec, p = .04), acceleration (524 +/- 261 cm/sec2 vs 199 +/- 111 cm/sec2, p < .001), and resistive index (88 +/- 12% vs 73 +/- 11%, p = .002). Differences were also noted between the group without significant arterial disease and the group with arteriolar dysfunction. When peak systolic velocity of less than 25 cm/sec or acceleration less than 400 cm/sec2 was used as an indication of inadequate arterial patency, the sensitivities were 35% and 100%, the specificities were 61% and 46%, and the negative predictive values were 42% and 100%, respectively, in the diagnosis of arterial insufficiency. CONCLUSION: Duplex sonography of the cavernosal arteries may be a useful screening tool in patients with suspected arteriogenic impotence only when acceleration is evaluated in addition to peak systolic velocity. The specificity of the method may be partially limited by the inability to distinguish between arterial and arteriolar disease. PMID- 8416651 TI - Correlation of duplex sonography with arteriography in patients with erectile dysfunction. AB - OBJECTIVE: Our objective was to assess the accuracy of using measurements of peak systolic velocity in the cavernosal artery for the diagnosis of arteriogenic impotence. MATERIALS AND METHODS: Twenty consecutive men with erectile dysfunction had duplex sonography after intracavernosal injection of papaverine to induce an erection. Peak systolic velocities in the right and left cavernosal arteries were measured by using Doppler sonography. Right and left selective penile arteriography was performed with low-osmolality contrast media after intracavernosal injection of papverine and intraarterial tolazoline. On the basis of the angiographic findings, penile arterial function was classified as normal, moderately insufficient, or severely insufficient. Doppler measurements of peak systolic velocity were correlated with arteriographic results. RESULTS: All 11 cavernosal arteries with peak velocities less than 25 cm/sec were associated with arterial disease, nine severe and two moderate. Thirteen of 17 carvernosal arteries with peak systolic velocities 25-34 cm/sec were associated with arterial disease, five severe and eight moderate. Only one of the 12 cavernosal arteries with peak velocity at or greater than 35 cm/sec was associated with arterial disease. CONCLUSION: We conclude that peak systolic velocity in the cavernosal artery as measured on duplex sonography is an accurate predictor of arterial disease in patients with erectile dysfunction. A peak systolic velocity of at least 35 cm/sec indicates normal arterial supply. At peak systolic velocities less than 35 cm/sec, the likelihood and severity of arterial disease increase as the peak systolic velocity decreases, with a peak velocity less than 25 cm/sec indicating a high likelihood of severe arterial disease. PMID- 8416652 TI - Transitional cell carcinoma. PMID- 8416653 TI - Imaging of en bloc renal transplants: normal and abnormal postoperative findings. AB - OBJECTIVE: Cadaveric kidneys from donors less than 5 years old, previously considered inferior graft material, are now being successfully transplanted en bloc into children and adults. On the basis of our experience with 132 patients, we describe the general principles of the procedure and review the spectrum of normal and abnormal imaging findings in patients who have undergone this promising transplantation procedure. MATERIALS AND METHODS: Paired cadaveric kidneys obtained from donors less than 5 years old (mean age, 24 months) were transplanted en bloc to 132 patients (mean age, 37 years) at our institution between 1981 and 1991. All available medical, surgical, pathologic, and imaging records were retrospectively reviewed to define the surgical technique, 1-year survival rate of the graft, appearance of the transplant on postoperative imaging studies, and the prevalence of and imaging findings caused by vascular, urinary, infectious, and neoplastic complications after transplantation. Complications were confirmed by a definitive imaging study, surgical exploration, or study of a pathologic specimen. RESULTS: Paired donor kidneys were transplanted en bloc extraperitoneally into the recipient's right or left iliac fossa, with intact portions of the donor aorta and inferior vena cava anastomosed to the recipient's external iliac artery and vein. One-year graft survival was 70% during the first 8 years of the study and 78% during the last 2 years. Postoperative imaging, particularly sonography and scintigraphy, clearly depicted the normal individual kidneys, urinary collecting systems, and en bloc vasculature. Postoperative complications were vascular (arterial stenoses and thromboses, venous thromboses, and pseudoaneurysms) in 18%, urinary (obstruction and anastomotic leak) in 11%, infectious (caliceal fungal balls) in 1%, and neoplastic (posttransplant lymphoma) in 1%. The complications involved one kidney in 60% of the patients and both kidneys in 40%. The imaging findings caused by these complications were similar to those caused by complications occurring after transplantation of single cadaveric kidneys; however, their detection was more difficult because of the complexity of the en bloc graft. CONCLUSION: Because of the shortage of available donor organs, en bloc renal transplantation will most likely become increasingly popular. Familiarity with the imaging appearance of the normal transplant and of posttransplantation complications will allow radiologists to perform effective postoperative imaging evaluations. PMID- 8416654 TI - Kidney dimensions at sonography: correlation with age, sex, and habitus in 665 adult volunteers. AB - OBJECTIVE: The purpose of this study was to investigate the normal sonographic measurements of the kidney in adult volunteers. SUBJECTS AND METHODS: Length, width, and thickness of the kidney and its central echogenic area and the parenchymal thickness of the upper pole were measured in an age- and sex stratified random sample of 665 volunteers 30, 40, 50, 60, and 70 years old. Measurements were made with the volunteers prone. Volumes of the kidney, the central echogenic area, and the renal parenchyma were calculated. Renal dimensions and renal and parenchymal volume were correlated with age, height, weight, body mass index, and total body area. In 94 subjects, renal length was measured with the volunteers supine also. RESULTS: Median renal lengths were 11.2 cm on the left side and 10.9 cm on the right side. Median renal volumes were 146 cm3 in the left kidney and 134 cm3 in the right kidney. Renal size decreased with age, almost entirely because of parenchymal reduction. Renal volume correlated best with total body area. Renal length correlated best with body height. Measurements of renal length obtained with the subjects supine were not significantly different from those obtained with the subjects prone. CONCLUSION: The most exact measurement of renal size is renal volume, which showed the strongest correlation with height, weight, and total body area. Clinically, measurement of renal length is most practical and can be done with the subject prone or supine. PMID- 8416655 TI - Nongynecologic applications of transvaginal sonography. AB - Transvaginal sonography has become an invaluable technique for examining the uterus and adnexa, primarily because it provides better spatial resolution than transabdominal sonography does. This characteristic also makes it useful for evaluating the nongynecologic structures in the pelvis. As many of these structures are imaged incidentally during gynecologic transvaginal sonography, familiarity with their normal and abnormal appearances is important. With minor modifications in technique, targeted studies of these structures are easily performed. We illustrate the technique used, normal anatomy seen, and abnormalities commonly encountered in transvaginal sonography of nongynecologic pelvic structures. PMID- 8416656 TI - Trauma to the upper thoracic spine: anatomy, biomechanics, and unique imaging features. AB - This review summarizes the anatomic and biomechanical features of the thoracic spine, which are different from those of the more mobile segments of the spine, and emphasizes their role in trauma. The distinguishing characteristics of the thoracic spine are the presence of the ribs and their articulations. The rib cage restricts motion and adds stiffness to the spine. During trauma, it provides the thoracic spine with additional strength and energy-absorbing capacity. Above the T10 level, most injuries produce a basic pattern consisting of an anterior fracture-dislocation involving two contiguous vertebrae, often with associated neurologic impairment. The definition of spinal instability remains controversial. CT is the imaging technique of choice for evaluation of spine fractures; however, MR imaging is superior in the evaluation of spinal cord injury and posttraumatic disk herniation. MR imaging also provides prognostic information not obtainable with other imaging methods. PMID- 8416657 TI - SI or bust! PMID- 8416658 TI - Early changes of body composition in human immunodeficiency virus-infected patients: tetrapolar body impedance analysis indicates significant malnutrition. AB - Total body water, body fat, body cell mass (BCM), extracellular mass (ECM), and the ECM-BCM ratio by impedance analysis were determined in 193 human immunodeficiency virus (HIV)-infected patients and 340 control subjects. Walter Reed (WR) classification was WR 2 in 26, WR 3-5 in 85, and WR 6 in 82 patients. Whereas resistance was increased, reactance and the phase angle were significantly reduced in all patient groups. Neither body weight nor body mass index (BMI) was affected in WR 2 patients, but BCM was reduced (31.9 +/- 4.3 vs 35.8 +/- 7.3 kg, P < 0.007) and ECM (31.2 +/- 4.4 vs 28.8 +/- 3.8 kg) as well as ECM-BCM ratio increased (0.99 +/- 0.14 vs 0.83 +/- 0.16, P < 0.001). In contrast to WR classification, diarrhea did not correspond with malnutrition. A loss of BCM (malnutrition) occurred already in otherwise symptomless HIV-infected patients (WR 2). This effect can be measured by tetrapolar impedence analysis but not by body weight or BMI. PMID- 8416659 TI - Forty years of the American Journal of Clinical Nutrition. PMID- 8416660 TI - Differential health benefits of weight loss in upper-body and lower-body obese women. AB - Upper-body obesity (UB Ob) is more strongly associated with adverse health consequences; however, few obesity-treatment studies have examined outcome according to body-fat distribution. To examine whether diet and formal- or informal-exercise instruction causes differential changes in health and lipid profiles, ten LB Ob and nine UB Ob premenopausal women received dietary intervention (2.1 MJ-deficit/d for 16 wk) and were randomly assigned to either formal- or informal-exercise instruction. Weight loss was similar between groups (approximately 8 kg), and no change occurred in lean body mass or basal metabolic rate. Baseline cholesterol and triglycerides were greater (P < 0.01) in UB Ob than LB Ob women and decreased more (P < 0.01) in response to treatment in UB Ob women. Formal exercise instruction increased high-density-lipoprotein cholesterol (P < 0.05) especially in UB Ob women. Future studies on treatment of obesity should include consideration of regional fat distribution. PMID- 8416661 TI - Dietary carbohydrate, muscle glycogen, and exercise performance during 7 d of training. AB - The effects of moderate- or high-carbohydrate diets on muscle glycogen and performance in runners and cyclists over 7 consecutive days of training were determined. Muscle biopsies were performed on 4 separate days before exercise for 1 h at 75% peak oxygen consumption (VO2) followed by five, 1-min sprints. After the training session on day 7, subjects ran or cycled to exhaustion at 80% peak VO2. Muscle glycogen for cyclists and runners was maintained with the high carbohydrate diet but was reduced 30-36% (P < 0.05) with the moderate carbohydrate diet. All subjects completed all training sessions, and there were no differences in times to exhaustion on day 7. For cyclists and runners, consuming a moderate-carbohydrate diet over 7 d of intense training reduces muscle glycogen but has no apparent deleterious effect on training capability or high-intensity exercise performance. A high-carbohydrate diet maintains muscle glycogen, but this has no apparent benefit on training capability or high intensity exercise performance. PMID- 8416662 TI - Energy expenditure in obese women before and during weight loss, after refeeding, and in the weight-relapse period. AB - In 10 moderately obese women, 24-h energy expenditure (24EE) was measured in a respiration chamber under four conditions: 1) before weight loss (body weight = 77.9 kg), 2) during weight loss (63.9 kg), 3) after realimentation (62.5 kg), and 4) 6-15 mo after the study diet with ad libitum diet (67.7 kg). The 14 +/- 8 kg (mean +/- SD) weight loss produced a decrease in 24EE of 1498 +/- 1138 kJ/d (P < 0.001), ie, a decrease of weight of 107 kJ.kg body wt-1.d-1. The subsequent 24EE (conditions 3 and 4) remained lower than the value before weight loss. A significant correlation was found between changes before and after weight regain in basal respiratory quotient (RQ) and the spontaneous rate of body-weight gain after cessation of the period of low energy intake (r = 0.89, P < 0.01); this suggests that the value of the postabsorptive RQ may be a predictor of relapse of weight gain. After discontinuation of the low energy diet, an elevated postabsorptive RQ shows that the endogenous lipid oxidation is low, a condition favoring weight gain. PMID- 8416663 TI - Changes in dietary intake, urinary nitrogen, and urinary volume across the menstrual cycle. AB - The effect of four menstrual cycle phases on changes in dietary intake, urinary nitrogen excretion, and urinary volume was examined in nine women confined to a metabolic unit, maintained at a constant activity level, and fed an ad libitum, rotating, staff-weighed diet. No significant changes in intakes of energy, protein, and fat occurred throughout the menstrual cycle although significant changes were found for intakes of ascorbic acid and water in food. A significant increase in consumption of carbonated, sugar-containing beverages was found in the luteal phase as was a significant increase in urine volume. Results suggest food choices and urine volume may be responsive to physiological regulators associated with hormonal changes in the menstrual cycle. Findings also suggest that energy intake is not altered across the menstrual cycle when physical activity is controlled and an accurate dietary assessment method is employed. PMID- 8416664 TI - Vitamin B-12, vitamin B-6, and folate nutritional status in men with hyperhomocysteinemia. AB - We measured the vitamin B-6, vitamin B-12, and folic acid nutritional status in a group of apparently healthy men (n = 44) with moderate hyperhomocysteinemia (plasma homocysteine concentration > 16.3 mumol/L). Compared with control subjects (n = 274) with normal plasma homocysteine (< or = 16.3 mumol/L) concentrations, significantly lower plasma concentrations of pyridoxal-5' phosphate (P < 0.001), cobalamin (P < 0.001), and folic acid (P = 0.004) were demonstrated in hyperhomocysteinemic men. The prevalence of suboptimal vitamin B 6, B-12, and folate status in men with hyperhomocysteinemia was 25.0%, 56.8%, and 59.1%, respectively. In a placebo-controlled follow-up study, a daily vitamin supplement (10 mg pyridoxal, 1.0 mg folic acid, 0.4 mg cyanocobalamin) normalized elevated plasma homocysteine concentrations within 6 wk. Because hyperhomocysteinemia is implicated as a risk factor for premature occlusive vascular disease, appropriate vitamin therapy may be both efficient and cost effective to control elevated plasma homocysteine concentrations. PMID- 8416665 TI - Brush border calcium uptake in short-bowel syndrome in rats. AB - Resection of distal small intestine causes calcium malabsorption in humans and in a rat model of 50% distal resection. We tested the hypothesis that this calcium malabsorption is caused in the rat model by a brush border defect. We compared brush border membrane vesicles from the proximal small intestine of control (transection and anastomosis at mid-small intestine) with distally resected rats. Mucosal protein was 25% greater in the resected group and the vesicles were enriched 37-fold in sucrase activity when compared with homogenate. Kinetic constants Vmax (maximal initial rate of saturable calcium uptake at infinite concentration), kT (calcium concentration for saturable calcium uptake rate at half Vmax), and KD (rate constant for nonsaturable calcium uptake per unit concentration) were slightly but not significantly greater in the resected as compared with the transected group, ruling out the brush border as the cause for decreased transmucosal calcium transport. PMID- 8416666 TI - The effect of fish oil on blood pressure in mild hypertensive subjects: a randomized crossover trial. AB - We conducted a double-blind, crossover trial with 18 healthy, untreated mildly hypertensive subjects to test the effect on blood pressure of 6 or 12 g fish oil/d (50% n-3 fatty acids) as compared with an olive oil placebo. Blood pressure was measured every 6 wk in the clinic and three times daily by subjects using a semiautomated device in their homes. Compliance was determined biochemically. No significant changes in home or clinic blood pressure measurements were noted for either dose after 6 or 12 wk of treatment. Clinic blood pressure after 12 g fish oil/d was slightly lower than after placebo treatment by -0.8/-0.4 mm Hg [95% CI: systolic blood pressure (-4.4, +2.8); diastolic blood pressure (-3.2, +2.4)]. Blood pressure changes were not correlated with compliance, baseline dietary fish consumption, or blood pressure. Moderate doses of fish oil did not have a substantial effect on blood pressure. We conclude that fish oil is not a practical treatment for mild hypertension. PMID- 8416667 TI - Evaluation of immunocompetence and nutritional status in patients with bulimia nervosa. AB - Nutritional status of patients suffering from bulimia nervosa was evaluated by anthropometric, hematological, and immunological parameters in comparison with a healthy control group. Anthropometry showed no signs of malnutrition in either group. Bulimic patients showed a significant decrease in most of the routine clinical indexes related to red blood cells. Lymphocyte subsets CD2 and CD4 in bulimic patients were lower than in controls, whereas CD8 remained unmodified, leading to a lower CD4:CD8 in bulimic patients. The decreases in CD2 and CD4 numbers might be linked to an impaired cell-mediated immune function, as evaluated by the response to skin tests. The B lymphocyte subset was increased in the bulimic group. Innate immunity was impaired in the bulimic patients. Although anthropometric and most of the routine clinical parameters were within the normal range, these results might suggest that the depleted immune function accounts for a subclinical malnutrition status in bulimic patients. PMID- 8416668 TI - Percentage distribution of fatty acids in subcutaneous adipose tissue of patients with peptic ulcer disease. AB - Dietary linoleic acid has been implicated in the pathogenesis of peptic ulcer disease because its metabolite arachidonic acid may be converted to cytoprotective prostaglandins. In addition, it has been suggested that the falling incidence and virulence of duodenal ulcer disease is related to increased dietary polyunsaturated essential fatty acid intake. In the present study the percentage content of linoleic acid in subcutaneous adipose tissue microbiopsies were used to see whether changes in percentage of fatty acids correlate with the presence or absence of an ulcer in individual patients. No significant difference in the adipose tissue content of linoleic acid was found, ie, 11.2 +/- 0.7% (n = 15) vs 10.9 +/- 0.5% (n = 15) in patients with peptic ulcer disease and matched control subjects, respectively. PMID- 8416669 TI - Marked resistance of normal subjects to tube-feeding-induced diarrhea: the role of magnesium. AB - The tolerance of healthy subjects to increasing rates of tube feeding was studied to better understand the etiology of diarrhea among tube-fed patients. Five volunteers were fed Osmolite HN by continuous duodenal infusion beginning at 314 kJ (75 kcal).kg body wt-1 x d-1 and progressing each 24 h until no longer tolerated. The five subjects were able to tolerate maximum 24-h infusions of 331 511 kJ.kg-1 x d-1 (198-340 mL/h). Diarrhea developed in only three subjects. Compared with nondiarrheal stools, the high-speed supernatant of the diarrheal stools had significantly higher concentrations of magnesium (192 +/- 22 mmol/L vs 139 +/- 17 mmol/L, P = 0.005), lower concentrations of potassium and phosphorus, and similar concentrations of calcium. The mean carbohydrate, fat, and nitrogen contents were not significantly different. We conclude that normal adult males are remarkably tolerant to duodenal infusion of this typical, isotonic tube feeding product. The diarrhea that occurred in three of the volunteers at very high infusion rates appeared to be osmotic and attributable predominantly to magnesium. PMID- 8416670 TI - Physical activity is a significant predictor of body density in women. AB - The purposes of this study were to 1) develop and cross-validate a prediction equation for body density with leisure-time physical activity (LTA) serving as a potential predictor, and 2) determine whether the addition of LTA to previously published equations would improve their prediction. Body density (hydrostatic weighing) was measured in a validation group of 111 women (aged 18-81 y) and was predicted by age, mass, body mass index (BMI), abdominal skinfold thickness, thigh skinfold thickness, and LTA. This equation was cross-validated on an independent group of women (n = 56). Consequently, groups were combined (n = 167) to yield a new equation that also included age, mass, BMI, abdominal skinfold thickness, thigh skinfold thickness, and LTA (R = 0.92, SEE = 7.9 g/L, P < 0.0001). LTA improved the prediction of equations published on women with relatively low body densities, but not on women with higher densities. In conclusion, LTA is an independent predictor of body density in healthy women and its prediction becomes stronger as body density decreases. PMID- 8416671 TI - Stable-isotope-tracer studies of amino acid balance and human indispensable amino acid requirements. PMID- 8416672 TI - Egg consumption and serum cholesterol. PMID- 8416673 TI - Fructose and blood cholesterol. PMID- 8416674 TI - Slipping to scapegoating in international nutrition. PMID- 8416675 TI - TSH beta subunit gene expression in human lymphocytes. AB - Production of neuroendocrine peptides by human lymphocytes is thought to facilitate control of the immune response. The presence of neuroendocrine peptide gene expression, specifically the TSH beta subunit gene, was studied in human lymphocytes using Northern blot analysis and polymerase chain reaction (PCR) techniques. Northern blot analysis of human lymphocyte RNA probed with a TSH beta cDNA probe failed to demonstrate TSH beta subunit steady state message levels. PCR amplification of lymphocyte-derived cDNA using TSH beta subunit complementary primers resulted in amplification of a .38 Kb DNA fragment, confirming expression and initial exonic splicing of TSH beta subunit gene exons 2 and 3 in human lymphocytes. Sequence analysis of this .38 Kb DNA fragment demonstrated conservation of exon borders after splicing (exons 2 and 3) and predicted an amino acid translation product similar, if not identical, to human TSH beta peptide sequence. Hybridization with a TSH beta subunit cDNA probe of PCR amplified reverse-transcribed lymphocyte RNA suggested that: (1) the abundance of TSH beta subunit gene transcripts in human lymphocytes is less than the relative abundance in T3-treated pituitary; and (2) this messenger RNA may be modulated by the presence of certain thyromimetic compounds (T2, T3, TRIAC). PMID- 8416676 TI - Ethanol enhances desialylation of transferrin by liver endothelial cells in the rat. AB - Desialylated transferrin is emerging as a reliable index of alcoholism, and liver endothelial cells are known to partially desialylate transferrin. The effect of a single intraperitoneal injection of ethanol on the desialylation of transferrin in the rat was studied. In pulse-chase experiments, fully sialylated diferric transferrin labeled with 125I (protein moiety) or 3H (sialyl residues) was incubated with isolated, fractionated liver endothelial cells from rats that were given ethanol. The supernatants then were subjected to column chromatography using RCA120-agarose to separate and quantify the desialylated fraction of transferrin. Saline-treated animals served as controls. Incubation of the endothelial cells derived from the liver of ethanol-treated rats resulted in a 263% increase in desialylation level of transferrin. Our data indicate that administering ethanol in rats enhances the desialylation of transferrin by rat liver endothelial cells. PMID- 8416677 TI - Lepromin skin testing in the classification of Hansen's disease in the United States. AB - Hansen's disease, or leprosy, although a relatively uncommon disease in the United States, continues to be important because of its implications--physical, psychological, and social--for the patient. Prognosis and treatment of the disease are based largely on clinical classification, which ranges from the multibacillary "lepromatous" to the paucibacillary "tuberculoid" forms, depending on the patient's specific immune capabilities. Traditionally, skin testing with lepromins--suspensions of the etiologic agent of Hansen's disease, Mycobacterium leprae--have been used as adjuncts to clinical parameters for classification in endemic areas. However, these have not been systematically studied in the United States. This report describes the results obtained from skin testing 38 volunteers (22 patients and 16 uninfected persons) with standard lepromin preparations. These results support the adjunctive value of lepromins for clinically classifying Hansen's disease in our "hypoendemic" population. PMID- 8416678 TI - Case report: azotemia secondary to enalapril and diuretic use and the diagnosis of renovascular hypertension. AB - When administered to a 45-year-old woman who was ill with severe hypertension, combined enalapril and hydrochlorothiazide therapy resulted in acute renal failure, leading to a successful search for renal artery stenosis. Acute renal failure as an adverse reaction to enalapril therapy occurs in approximately 20% of patients with secondary hypertension from renovascular disease. On the other hand, in a group of essential hypertensives this occurs at a rate of approximately 0.2%. Thus, a likelihood ratio of 100 is produced, and given the prevalence of renovascular hypertension of approximately 2% in all hypertensives, a posterior probability of 0.67 can be calculated. The occurrence of this adverse effect should indicate that a diagnostic work-up to rule out renovascular hypertension in such patients should be pursued. The sensitivity and specificity of this "test" are 0.20 and 0.998, respectively. This illustrates that the mathematic aids in decision theory need not be applied solely to those more traditional aspects of diagnostic testing, but may be useful in other situations as well. PMID- 8416679 TI - Case report: cerebral venous thrombosis as a manifestation of acute ulcerative colitis. AB - Ulcerative colitis is associated with a number of extraintestinal complications, including the infrequent occurrence of thromboembolic disease. Cerebral venous thrombosis is extremely rare, with only 15 cases reported in the literature. The outcome in this group of patients is poor; the result in more than 80% of the cases is permanent neurologic sequelae or death. The precise mechanisms involved in thrombogenesis remain unclear. A hypercoagulable state may occur in ulcerative colitis because of well-documented associations of thrombocytosis, elevated factors V and VIII and fibrinogen, and decreased antithrombin III. Treatment regimens for cerebral venous thrombosis remain controversial and include anticoagulation with heparin, surgical thrombectomy, and systemic and local infusion of fibrinolytic regimens. A conservative approach with antiedemic agents, anticonvulsants, antiplatelet therapy, and acetazolamide also may be beneficial and offers a substantially reduced hemorrhagic potential. A case of primary superior sagittal sinus thrombosis associated with active ulcerative colitis treated by the latter method is reported. The patient's neurologic recovery was complete, with recanalization of the thrombosed venous sinus being demonstrated by magnetic resonance imaging studies. PMID- 8416680 TI - Case report: acyclovir neurotoxicity and nephrotoxicity--the role for hemodialysis. AB - Severe neurotoxicity and acute renal failure developed in a patient with newly diagnosed AIDS while receiving high-dosage intravenous acyclovir for disseminated herpes zoster. Hemodialysis resulted in a rapid resolution of neurologic symptoms and was associated with a reduction in plasma acyclovir concentration. Acute hemodialysis therapy should be considered in cases of serious neurotoxicity secondary to acyclovir, especially when accompanied by renal failure. PMID- 8416681 TI - New approaches to the therapy of autoimmune diseases: rheumatoid arthritis as a paradigm. AB - Several therapeutic agents currently are used to treat rheumatoid arthritis (RA). However, there is no compelling evidence that any of these agents substantially alters the long-term destructive course of RA. Advances in biotechnology have led to a better understanding of mechanisms that underlie autoimmune diseases such as RA. Although the etiology of RA remains unknown, there now is considerable insight regarding the immune and inflammatory pathways that ultimately lead to cartilage and bone destruction. Therapies with monoclonal antibodies directed against cell surface constituents, fusion toxins against cell activation markers, and cytokine inhibitors all have been shown to be safe and possibly efficacious in early open trials in RA. They now are being more rigorously tested in double blind, placebo-controlled trials. Early experience with these biologic agents in humans, as well as data obtained from the use of these agents in animal models of autoimmune disease, are reviewed. In addition, experimental studies with "blocking peptides" and immunization with autoreactive T cell receptor peptides will be reviewed, and implications for therapy in RA will be discussed. PMID- 8416682 TI - Tuberculosis--immunopathogenesis and therapy. AB - Infection with Mycobacterium tuberculosis (TB) has returned to the forefront of public and medical concern because of the recent sharp increase in the number of cases. Major strides have been made in understanding the pathogenesis of TB, and some of these basic advances are being applied clinically. This review focuses on current concepts of the host response to TB, the changing epidemiology of TB, and optimal treatment strategies. PMID- 8416683 TI - Measurement of phosphocreatine in cutaneous tissue by high pressure liquid chromatography. AB - A high pressure liquid chromatography technique for measuring phosphocreatine and adenine nucleotides in human cutaneous tissue is described. The presence of phosphocreatine in human skin was confirmed by this analytic method. Molar concentration of phosphocreatine and adenine nucleotides were determined in normal skin and in benign and malignant skin lesions. The preliminary results suggest that absolute amounts of phosphocreatine and adenine nucleotides and phosphocreatine/adenosine triphosphate ratios in malignant skin neoplasms and benign cutaneous lesions are different from those measured in normal nonischemic human skin. PMID- 8416684 TI - Fast spin-echo imaging of the brain. AB - Fast spin-echo (FSE) imaging is a recently modified rapid-acquisition relaxation enhanced technique that affords rapid magnetic resonance (MR) imaging while retaining true spin-echo (SE) contrast features. By manipulating factors such as echo train length, echo spacing, and order of phase encoding, dual-echo brain images currently may be obtained many times faster than with conventional SE techniques. The time advantage of FSE may be used simply to obtain images much more rapidly or to acquire images of much higher contrast or spatial resolution compared with conventional SE images in a comparable period of time. Lesion appearance and conspicuity are usually quite similar to those on conventional SE images, with the important exception of some hemorrhagic blood products, which may appear less conspicuous on FSE images because of reduced magnetic susceptibility effects. Another important difference is the bright appearance of fat on all routine FSE sequences (T1-weighted, proton-density, and T2-weighted), although this may be eliminated easily by using routine fat-suppression techniques. Recent applications of FSE include three-dimensional multislab imaging and high-resolution imaging with large-matrix, small-field of view, thin slice, and multiple-excitation imaging parameters. FSE techniques appear quite promising for rapid MR imaging of the brain, and they have largely replaced conventional SE technique at some institutions. PMID- 8416685 TI - Ultrafast imaging of brain tumors. AB - Ultrafast magnetic resonance (MR) imaging allows the acquisition of anatomic images of the brain in a fraction of a second. Recently developed imaging techniques have the potential to increase the specificity of information provided by MR imaging. MR imaging techniques that are based on ultrafast scanning have enabled the collection of functional as well as anatomic information concerning tumor microvasculature. These techniques promise to increase knowledge of the tissue characteristics of human brain tumors, their progress, and their responses to therapy. PMID- 8416686 TI - Diffusion and perfusion magnetic resonance imaging in brain tumors. AB - The article reviews recent progress made in the field of diffusion and perfusion magnetic resonance (MR) imaging and presents possible applications in brain tumors. Diffusion, a new parameter, provides useful data to assess tissue structure and function. Perfusion MR imaging gives results that are somewhat similar to those obtained with classic non-MR imaging methods, but it offers several potential advantages. PMID- 8416687 TI - In vivo magnetic resonance spectroscopy of human brain tumors. AB - In vivo nuclear magnetic resonance (NMR) spectroscopy is a rapidly developing, noninvasive analytical technique that allows sequential studies of brain tumor metabolism. It can be implemented on most conventional high-field magnetic resonance imaging scanners. In animal tumor models, NMR spectroscopy has been used extensively to characterize the metabolic changes associated with tumor growth and response to therapy. Preliminary data in humans indicate the feasibility of recording phosphorus and proton spectra from focal lesions within the brain. It is hoped that the technique will provide useful prognostic information in terms of tumor growth behavior and prediction and detection of response to therapy. However, the technique suffers from relatively coarse spatial resolution, and the methodology is still under development. More studies with larger patient groups are required before the clinical utility of the technique can be fully evaluated. PMID- 8416688 TI - Magnetic resonance scanning technique for neuroimaging in pediatrics. AB - This article reviews the magnetic resonance imaging technique employed in children. It discusses preparing children for the examination, sedation techniques, imaging sequences, and the use of contrast media. PMID- 8416689 TI - Magnetic resonance angiography in pediatric neuroradiology. AB - Magnetic resonance angiography (MRA) is a novel technique that uses gradient echo pulse sequences and computer postprocessing technology to create vascular-flow images. The technique is easy to apply, noninvasive, and frequently yields information not available by other noninvasive means. The utility and benefits of MRA are amply documented by clinical studies in the neuroimaging literature. This article reviews the techniques of MRA and the current literature in the application of MRA in pediatric imaging. Illustrative cases demonstrating the use of MRA in pediatric neuroradiology are presented. The utilization of MRA in congenital lesions such as aberrant internal carotid artery and arteriovenous malformation and in differing causes of ischemic disease is highlighted. There is an ever-expanding number of applications of MRA in pediatric neuroimaging. Despite the lack of standardized protocols, with a thorough knowledge of the basic principles, MRA can be successfully applied in many imaging situations. The ease of use and noninvasive nature of MRA make it an ideal tool in the evaluation of the pediatric neurovasculature. PMID- 8416690 TI - [Incidence rates of cerebral palsy, severe mental and motor retardation, and Down syndrome in the city of Kokubunji in suburban Tokyo]. AB - We investigated the incidence rates of cerebral palsy, severe mental and motor retardation, and Down syndrome in the City of Kokubunji in suburban Tokyo with a total population of about 100,000. The number of liveborn babies during 1985-1989 was 5,475. The number of children with cerebral palsy (CP) was 11 and the incidence rate was 2.01/1,000, which was equal to the rates in reports from several countries and was higher than those of the previous reports from Japan. Two out of 10 patients with CP had mild motor handicaps and were expected to "outgrow" their handicaps. The number of children with "severe mental and motor retardation" (SMMR) was 6 and the incidence rate was 1.10/1,000. Prenatal brain damage played a major role in the pathogenesis of CP and SMMR. The number of children with Down syndrome (DS) was 11 and the incidence rate was 2.01/1,000 which was higher than the previous rates. Incidence rates of CP, SMMR, and DS still remain high and further strategy to prevent pediatric neurological diseases is necessary. PMID- 8416691 TI - [Electrophysiological study of Bell's palsy in children]. AB - An electrophysiological study of Bell's palsy in children was reported. Direct response to facial nerve stimulation was tested in 28 patients. Of these, 17 patients were further tested for electrically elicited blink reflex. The reduction in amplitude of direct response on affected side was characteristic of Bell's palsy. The direct response was evoked on affected as well as nonaffected side. The amplitude of the direct response on the affected side was expressed as percentage of that on the nonaffected side (ratio of M response amplitude:RMA). 13 patients showed incomplete recovery after 3 months, and RMA of the 11 patients of them were reduced below 30%. In the patients showing good clinical recovery within 2 months, RMA returned to normal promptly. In the blink reflex, R1 R2 components were found to be persistent on the first examination, suggesting a satisfactory functional outcome in Bell's palsy. As these electrophysiological methods are conventional assessing facial nerve function, it seemed to be valuable for the follow up of children with Bell's palsy. PMID- 8416692 TI - [Long-term clinical course of sequelae in patients with neonatal anoxic encephalopathy resulting in profound mental retardation and motor disturbance]. AB - A long-term observation has been made in 58 patients (30 males and 28 females) with severe sequelae of neonatal anoxic encephalopathy. They aged from 8 months to 65 years. All of them had motor disturbances and profound mental retardation. Motor function was improved in 4 patients with aging. In contrast, motor activity deteriorated in 11 cases, of which 4 showed a mental regression. Among them, patients who had originally better motor ability than sitting were likely to deteriorate by uncontrollable epilepsy and/or excessive administration of anticonvulsants. Regression of the patients with worse motor ability like bedridden appeared to attributable hypertonia of muscles and bodily deformation. Fifteen cases showed an exacerbation of general condition which originated predominantly to respiratory distress. Twelve patients died including 6 exacerbated cases. Exacerbation or death may have occurred frequently in specific periods of infancy, adolescence and youth with the patients who showed very low motor function such as bedridden and no locomotion. PMID- 8416693 TI - [Posture in preterm infants is not a good indicator of brain damage]. AB - The development of posture during the prenatal period was studied longitudinally in 12 matched pairs of low-risk and high-risk preterm infants. The gestational age of the infants in both groups ranged from 28 to 36 weeks. The two members of each pair had the same gestational age at birth and the video recordings were performed at the same postmenstrual age. They were filmed more than three times during the prenatal period, each video recording lasting 1 h. Time of occurrence and duration of each posture were analyzed from replay of the video recordings. No age-specific preference posture described in the literature was found from birth until the term age between two groups. There was no age-related tendency in duration and frequency of postures between two groups. There was a clear-cut dominant head position to the right side from 31 weeks onward. Asymmetrical tonic neck postures (ATN) were very inconsistently present before the term age. And there was no significant difference in the occurrence of ATN postures between two groups. We could not find any specific abnormal postures in the high-risk group. Our study suggested that posture in preterm infants is not a good indicator of brain damage in the prenatal period. PMID- 8416694 TI - [Prognostic significance of auditory brainstem responses in full-term newborn infants with intracranial hemorrhages]. AB - Auditory brainstem response (ABR) was recorded in 41 full-term newborn infants with intracranial hemorrhages (ICH; 10 infants with and 31 infants without neurological sequela) and the data were compared with those obtained in normal full-term newborn infants as controls. The wave-I peak latency was significantly prolonged in the non-sequela group than in the control group and in the sequela group than in the non-sequela group. The wave-III and wave-V peak latencies were significantly prolonged in the ICH group (sequela and non-sequela groups) than in the control group but did not significantly differ between the sequela and non sequela group. The wave I-V interpeak latency did not significantly differ among the three groups. Among 10 infants in the sequela group, 9 had a V/I amplitude ratio (the amplitude of wave-I divided by the amplitude of wave- V) of less than 1.0. This suggests that the V/I amplitude ratio is of prognostic value in ICH infants. PMID- 8416695 TI - [Natural killer cell activity in severely handicapped children: correlation with malnutrition and frequent infections]. AB - We studied natural killer cell (NK) activity in 20 severely handicapped children and investigated the relationship between NK activity and malnutrition. We divided our patients into two groups with regard to frequency of infections: high frequency group (group 1) and low frequency group (group 2). NK activity in group 1 was low in comparison with group 2. Calorie intake and Rohrer's index were lower in group 1. Serum albumin (Alb) and iron (Fe) in group 1 were also lower than those in group 2, and serum zinc (Zn) and hemoglobin (Hb) were slightly lower in group 1. NK activity was positively correlated with calorie intake, Alb, Zn and Hb. Six patients with low NK activity were given supplementary diet; two patients were given protein-enriched diet; three patients were given protein enriched diet and zinc sulfate; one patient was given increased diet. NK activity rose in 4 patients. These results indicated that the depression of NK activity was associated with frequent infections and correlated with nutritional conditions of the severely handicapped children. PMID- 8416696 TI - [Long-term neurological outcome in children with convulsions during exanthema subitum]. AB - We examined long-term neurological outcome of 12 patients, whose first febrile seizures had occurred during exanthema subitum (ES) and who had been treated with continuous daily administration of anticonvulsant drugs. Six of 12 children had a family history of febrile or afebrile convulsions and in a sister of one patient, athetoid palsy followed the status of afebrile convulsion (AC) during ES. Twelve cases were clinically classified into four groups; (A) 2 patients in whom ACs were repeated as soon as the fever of ES had dropped, (B) 6 patients in whom frequent recurrences of febrile convulsion followed ES, showing epileptic EEG abnormalities, (C) 1 patient who had suffered from later epilepsy, (D) 3 patients who showed complex type of febrile convulsion only during ES without any EEG abnormalities. Except one patient in group (A), who showed residual hemiparesis on the left, 11 cases demonstrated normal psychomotor development, and in 5 of all 12 cases, the treatment with anti-convulsant drugs could be discontinued. Unless severe neurological sequelae are complicated, long-term prognosis is not so poor in children whose first febrile convulsion occurred during ES. PMID- 8416697 TI - [Prognosis of epilepsy in psychoneurologically normal children]. AB - Prognosis of 175 psychoneurologically normal children with the onset of epilepsy after 18 months of age were studied. The remission rate of partial epilepsy (81 of 107 cases, 76%) was higher than that of generalized epilepsy (34 of 56 cases, 61%). Prognoses for patients with sleep epilepsy (52 of 62 cases, 84%) were better than those for patients with waking epilepsy (63 of 101 cases, 62%). Most (95%) of the children with sleep epilepsy had partial epilepsy, including benign childhood epilepsy with centrotemporal spike (BECT, 36 cases) and partial epilepsy other than BECT (23 cases). Benign courses were not limited to BECT patients. Generalized waking epilepsy other than absence had the lowest remission rate. Eleven children exhibited mild mental retardation on last examination. Most of them had atypical absence and/or myoclonic seizures with the onset of seizure before 3 years of age. It appeared important to classify the epileptic children with regard to time of seizure occurrence as well as seizure type to determine their prognoses. PMID- 8416698 TI - [A female child with Down syndrome complicated by spinal cord compression by atlanto-axial dislocation]. AB - A 3-year-old girl with Down syndrome was admitted to our hospital, associated with hypotonia and dyspnea. She presented frog-posture and shallow breathing. The blood gas analysis revealed hypercapnic acidosis with pH 7.371, PO2 74.6 mmHg, PCO2 52.6 mmHg, and BE 3.5. The cervical X-ray films with flexion and extension of head showed anterior dislocation of the atlanto-axial articulation. Magnetic resonance imaging clearly showed a severe cord compression between C1 and C2. In children, spinal cord compression induced by dislocation of the atlanto-axial articulation is very rare. However, we should take into consideration of this insidious risk associated often with Down syndrome. PMID- 8416699 TI - [A case of multiple sclerosis associated with chronic inflammatory demyelinating polyradiculoneuropathy]. AB - We report a 19-year-old girl with multiple sclerosis (MS) who had chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). At the age of 9 (1981), she was diagnosed as having Devic disease, including optic neuritis and transverse myelitis. For two years after the onset of the disease, she suffered from three relapses of MS and was treated with prednisone. At the age of 11, she noticed muscle weakness and sensory disturbance of four limbs. On examination, all tendon reflexes were absent, CSF protein was elevated, and motor nerve conduction velocity of the ulnar nerves was markedly slowed. Sural nerve biopsy showed decreased density of myelinated fibers and de-re-myelinated fibers with onion-bulb formations. Between 11 and 19 years, she had a number of episodes of the central nervous system (CNS) and peripheral nervous system (PNS) deficits with partial or complete recovery by oral administration of prednisone. The clinical course and laboratory data are consistent with MS associated with CIDP. Such our case of MS with CIDP in childhood is very rare. Patients with MS and CIDP may represent a subpopulation who had a common pathogenetic factor for both CNS and PNS demyelination. PMID- 8416700 TI - [A case of degenerative type of progressive myoclonus epilepsy]. AB - This report describes a patient with degenerative type of progressive myoclonus epilepsy (PME), who showed slowly progressive deterioration of the central nervous system; intellectual impairment, dysarthria, and involuntary movements, particularly action myoclonus and dystonia. The patient was a 19-year-old woman who had no hereditary factors. At the age of 4, she developed action myoclonus in the upper limbs bilaterally. Her condition became gradually worse, and at the age of 15, she was admitted to our hospital because of involuntary movement in the upper limbs. First physical examination revealed mild mental retardation, action myoclonus, dystonia, and delayed adolescence. As giant SEP characteristic of PME and Ramsay Hunt syndrome was found, she was tentatively diagnosed as having Ramsay Hunt syndrome without epilepsy, and delayed adolescence. Now, she is 19 years old, and unable to walk alone because of involuntary movements and paralysis. But she has not developed epilepsy. As she has not been compatible with progressive myoclonus epilepsy (PME) and progressive myoclonic ataxia (PMA) classified by Marseille Consensus Group, she has been diagnosed as having an atypical PME syndrome. PMID- 8416701 TI - [A boy with Down syndrome and adrenoleukodystrophy]. PMID- 8416702 TI - [Serial MR imaging of possible Pelizaeus-Merzbacher disease]. AB - We performed serial magnetic resonance imaging (MRI) toward 4 male children with possible Pelizaeus-Merzbacher disease (PMD). They were clinically suspected as PMD on the basis of typical neurological, electrophysiological and MRI examinations. MRI revealed that myelinated fibers were localized in brainstem, cerebellum, internal capsule, optic radiation and proximal corona radiata in case 1, while myelinated fibers were almost absent in case 2-4. On repeated MRI investigation, we found no further extension of myelination in case 1 and the absence of myelination in case 2-4. This study demonstrated that myelination in the brain is arrested immediately after birth in case 1, while no myelination developed before birth in case 2-4. It was speculated that MRI in case 1 and case 2-4 might correspond to neuropathologic findings of classical and connatal form as reported in the literature. PMID- 8416703 TI - [Clinical evaluation of evoked potential examination for monitoring in multiple sclerosis]. PMID- 8416704 TI - [A case of Kearns-Sayre syndrome like mitochondrial encephalomyopathy with brain stem and cerebellar manifestations]. PMID- 8416705 TI - Cell kinetics and prognosis in gastric cancer. AB - METHODS: The potential proliferative activity of primary gastric cancer was determined using the in vitro tritiated thymidine labeling index (LI) technique. RESULTS: The proliferative rate had a wide range (0.1-28.4%) with a median value of 9.3%. The cell kinetics of the primary tumor were not related to clinicopathologic features, such as the patient's age and sex or the tumor's histologic type and stage. The contribution of cell kinetics to prognosis was investigated in a series of 28 patients (median follow-up, 34 months). The 3-year survival rate was 50% for patients with slowly proliferating tumors compared with only 13% for those with rapidly proliferating tumors. Moreover, in patients with high-LI tumors, the risk of death was more than sixfold greater than for those with low-LI tumors. CONCLUSIONS: These data suggest that cell kinetic studies might be an important discriminant to predict prognosis in gastric cancer. PMID- 8416706 TI - Leukemic infiltration of the esophagus. AB - BACKGROUND: Leukemic infiltrates of the esophagus have been described occasionally in autopsy series, but there are no reports of antemortem diagnosis. METHODS: Case reports are presented for three patients with acute myeloid leukemia in whom leukemic infiltration of the esophageal mucosa was diagnosed histologically and cytologically by endoscopic examination. Autopsies of patients with leukemia from 1976-1988 were reviewed. RESULTS: The autopsy review of 207 patients with leukemia showed evidence of leukemic infiltration in the esophagus in 7.2% of cases. The only clinical factor identified to be significantly associated with esophageal involvement by leukemic cells was a high initial leukocyte count. Esophageal involvement was associated with leukemic infiltration of other soft tissues and organs. CONCLUSIONS: Although the etiology of dysphagia in patients with acute leukemia is usually related to infection, reflux, chemotherapy toxicity, or benign strictures, the frequency of esophageal leukemic infiltration in this autopsy series suggests that this diagnosis must be considered. Esophageal leukemia is usually associated with widely disseminated soft tissue and visceral infiltrates. PMID- 8416707 TI - Acute neurotoxicity after intrathecal cytosine arabinoside in two adolescents with acute lymphoblastic leukemia of B-cell type. AB - BACKGROUND: Two adolescents with acute B-cell leukemia (Burkitt leukemia) had acute severe neurotoxicity after treatment with intrathecal (IT) cytosine arabinoside (AraC) at a dose of 50 mg/day for three consecutive days. RESULTS: A 16-year-old boy had a rapidly ascending myelopathy and encephalopathy 20 hours after receiving the third dose of IT AraC. He remained quadriplegic and required ventilatory assistance for 10 months until his death from progressive tumor. A 12 year-old girl had acute encephalopathy, seizures, and focal neuroimaging abnormalities in the cerebellum and brain stem within 32 hours of the third AraC dose and 8 hours after IT methotrexate (MTX, 12 mg). Her clinical neurologic deficits resolved during the ensuing month. Patient 1 represents the first report to the authors' knowledge of acute severe neurotoxicity after AraC administered as the only IT drug. In Patient 2, IT AraC neurotoxicity may have been potentiated by the single dose of MTX. CONCLUSION: IT AraC administered for 3 or more consecutive days may lead to profound neurologic dysfunction and require discontinuation of therapy. PMID- 8416708 TI - Specific skin manifestations in acute leukemia with monocytic differentiation. A morphologic and immunohistochemical study of 11 cases. AB - BACKGROUND: Monocytic differentiation is present in the myelomonocytic (M4) and monocytic (M5) type of acute myeloblastic leukemia. Infiltration of the skin in acute myelomonocytic leukemia occurs in 10-20% of patients, the skin lesions occasionally being the first symptom, even preceding monocytosis. METHODS: Eleven patients with myelomonocytic (n = 2) and monocytic leukemia (n = 9) were studied who had skin manifestations. RESULTS AND CONCLUSIONS: Clinically, all patients showed disseminated papules or nodules that corresponded histologically to nodular or diffuse infiltrates of monocytoid cells, occasionally displaying a whorled pattern. The currently available antibodies for paraffin-embedded sections (lysozyme, elastase, leukocyte common antigen (CD45), MT1 (CD43), Leu-M1 (CD15), LN2 (CD74), MB2, MB1 (CD45RA), LN1 (w75), Mac387, L26 (CD20), UCHL1 (CDR0), MT2 (CD45RA), and KP-1 (CD68)) and chloracetate-esterase are not more helpful in diagnosis than are the histologic findings. By contrast, the antibodies used on frozen sections (Leu-4 (CD3), Leu-3a (CD4), BA1 (CD24), B4 (CD19), Leu-M5 (CD11c), Vim12 (CD11b), VimD5 (CD15), KiM6 (CD68), KIM7 (CD68), My7 (CD13), and My9 (CD33) allow the definition of a reaction pattern that is diagnostic for acute myeloid leukemia with monocytic differentiation. PMID- 8416709 TI - Central nervous system involvement of adult T-cell leukemia-lymphoma with multinucleated giant cells in the brain, skin, and kidney. AB - BACKGROUND: A case of central nervous system (CNS) involvement in a patient with adult T-cell leukemia-lymphoma (ATLL) with multinucleated giant cells (MNGC) is presented. METHODS: A 48-year-old woman with human T-lymphotropic virus type I (HTLV-I) antibody titer had multiple focal brain symptoms and skin eruptions without lymphadenopathy, hepatosplenomegaly, or increased abnormal lymphocytes in the peripheral blood. No spastic paraparesis of the lower limbs was found. The encephalopathy was progressive, and she died 5 months later despite repeated intrathecal administration of methotrexate, cytosine arabinoside, and prednisolone and monthly systemic chemotherapy with doxorubicin, cyclophosphamide, vincristine, and prednisolone. RESULTS: Postmortem examinations identified unusual ATLL lesions composed of marked infiltrations of atypical mononuclear cells and bizarre MNGC with histiocytic granulomatous reactions in the leptomeninges, brain tissues along the Virchow-Robin spaces, skin, and kidney. Immunohistochemical stains confirmed the T-cell nature of such mononuclear cells and partially T-cell and partially macrophage nature of the MNGC, although no evidence of HTLV-I expression was found. CONCLUSIONS: ATLL presenting with CNS symptoms is rare. It was assumed that the direct cytopathic effects of HTLV-I were responsible for the formation of the MNGC after considering the similarity with MNGC in encephalopathy caused by the human immunodeficiency virus. PMID- 8416710 TI - Diagnosis of brain metastases from a primary hemangiosarcoma of the spleen with magnetic resonance imaging. AB - BACKGROUND: Primary malignant splenic neoplasms are rare tumors. Apart from involvement of the spleen by malignant lymphoma, hemangiosarcoma is the most common form of primary malignant neoplasm in the spleen. The diagnosis often is made at autopsy or after spontaneous splenic rupture. The prognosis is usually poor, and widespread metastases soon occur. METHOD: Currently, no case of metastasis to the brain has been reported to the authors' knowledge. RESULTS: This case report describes a patient who, as the first sign of metastatic disease, had symptoms from brain metastases 3 months after splenectomy. CONCLUSION: Magnetic resonance imaging was diagnostic. PMID- 8416711 TI - Stenting in malignant vena caval obstruction. AB - BACKGROUND: For patients with recurrent vascular obstruction of the vena cava due to tumor regrowth after chemotherapy or radiation therapy and occasionally surgery, no current therapy is available. With the development of intravascular stenting, a new option becomes available in the treatment of a vena caval syndrome. METHODS: Twenty-two patients were treated for malignant obstruction of the vena cava by single, double, and triple Z-type metal stents. All patients had been pretreated extensively and their disease was not amenable to other therapeutic techniques at the time of stenting. RESULTS: Correct positioning of the stents was achieved in all patients. In 68% of patients (n = 15), the symptoms completely disappeared without recurrence until death as a result of tumor progression. In 18% of patients (n = 4), a remarkable improvement was observed, but there was no complete disappearance of symptoms. In 14% of patients (n = 3), reocclusion of the vena cava occurred. The median survival time was 3.0 months (range, 1 week to 9 months). CONCLUSIONS: The application of Z-type metal stents in patients with recurrent malignant obstruction of the vena cava appears to be a useful palliative procedure. PMID- 8416712 TI - The prognosis of breast cancer in males. A report of 335 cases. AB - BACKGROUND: Because of the relative rarity of breast cancer in males, data have not been sufficient to support a definitive analysis of pertinent prognostic factors. Remarkably, no studies of male patients with breast cancer have presented survival information based on the number of histologically positive axillary nodes, the most sensitive single indicator of prognosis in women with breast cancer. METHODS: In this study, the clinical course of breast cancer was documented for 335 male patients registered from 1965 through 1986. For patients to be eligible, diagnoses had to be made within 3 months of registration and the patients had to have histologic confirmation and receive part or all of their initial treatment at 1 of 11 cancer centers participating in the International Patient Data Exchange System. RESULTS: The survival rate at 10 years was 84% for patients with histologically negative nodes, 44% for those with one to three positive nodes, and 14% for the group with four or more histologically positive nodes. The survival rates at 5 years were 90%, 73%, and 55%, respectively. In a multivariable analysis, the risk of death due to breast cancer for a patient with four or more histologically positive nodes was 6.75 times that of a patient with negative nodes. CONCLUSIONS: The findings of the authors indicate the following: (1) The number of histologically positive axillary nodes and, to a lesser degree, tumor diameter are significant prognostic factors for breast cancer in male patients. (2) The prognosis of breast cancer is the same in male and female patients when compared on the basis of the number of histologically positive nodes. PMID- 8416713 TI - Steroid hormone receptor immunohistochemistry and amplification of c-myc protooncogene. Relationship to disease-free survival in breast cancer. AB - BACKGROUND: It is important to develop parameters that aid in prognosticating which patients with breast cancer are more likely to have a rapid disease course and therefore might benefit from early aggressive therapies. METHODS: Specimens from two groups of women, deliberately selected because their clinical courses differed greatly, were studied to detect amplification of the protooncogenes c myc, int-2, and C-erbB-2/neu by slot-blot assay, the estrogen receptor (ER), and the progesterone receptor (PR) by both biochemical and immunohistochemical procedures (ERICA and PRICA). One group of 50 patients had a prolonged disease free interval after initial surgery (mean, 6.4 years); the other group of 52 women had had rapid disease recurrence (mean, 1.4 years) or progression (5 patients died of disease within 1 year of diagnosis). The patients were selected from 1700 consecutively accessioned cases if they fit the study criteria and sufficient tissue was available for oncogene hybridization studies. RESULTS: The two groups differed statistically by stage, number of involved axillary lymph nodes, ERICA and PRICA results (P = 0.001), and amplification of c-myc (P = 0.003). The percentage of patients with rapid disease recurrence and progression increased from 0-93% when risk factors changed from best case (ERICA and PRICA results, positive; c-myc, not amplified; and axillary nodes, not involved) to worst case (ERICA and PRICA findings, negative; c-myc, amplified; and axillary nodes, involved). CONCLUSIONS: Women with these worst-case parameters were more likely to have a recurrence sooner and rapidly progressive disease. They might benefit from early aggressive therapeutic measures. PMID- 8416714 TI - Survival of first and second primary breast cancer. AB - BACKGROUND: A second primary tumor (SPT) in the breast is the most common one seen in clinical practice. There are conflicting reports regarding the incidence and survival of patients with SPT in the breast. METHODS: To elucidate this, data on 139,932 patients with primary breast cancer, reported to the Surveillance, Epidemiology, and End Results Program between 1973 and 1986, were analyzed. Of these, 3431 patients had a contralateral metachronous breast cancer (interval, > 6 months). RESULTS: Survival from the date of diagnosis of the metachronous breast tumors was compared with that of patients with single breast tumors, controlling for age at diagnosis, stage, race, and treatment. In the multivariate analysis, age and stage at diagnosis of first and second tumors were the only variables that significantly influenced survival. The survival of patients with localized SPT was similar to that of patients with a localized single breast tumor. Patients with regionally advanced SPT lived for a shorter time than did corresponding patients with only one tumor. CONCLUSIONS: Patients with single breast cancer were at increased risk of having a SPT in the breast. As a result, efforts for early detection by physical examination and mammography of survivors of single breast cancer are advocated because the survival of patients with a localized SPT in the breast is as good as that of those with a single localized tumor. PMID- 8416715 TI - Type II estrogen binding sites and antiproliferative activity of quercetin in human meningiomas. AB - Eleven cases of meningiomas were investigated for the presence of estrogen and progesterone receptors. These tumors specifically bound estradiol. This binding activity almost exclusively resulted from the presence of high numbers of type II estrogen binding sites (EBS). Estrogen receptors were absent or present at low concentrations. Competition analysis showed that the flavonoid, quercetin, competed for tritiated estradiol binding to type II EBS; both rutin and hesperidin did not. In addition, 10 microM quercetin, unlike rutin or hesperidin, was effective in inhibiting in vitro bromodeoxyuridine incorporation by meningioma cells. Although the mechanism of the antiproliferative activity of quercetin remains to be clarified, it is possible that this flavonoid may regulate cell growth through a ligand interaction with type II EBS. PMID- 8416716 TI - Proliferative potential of brain tumors. Analyses with Ki-67 and anti-DNA polymerase alpha monoclonal antibodies, bromodeoxyuridine labeling, and nuclear organizer region counts. AB - Histochemical analyses with Ki-67 and anti-DNA polymerase alpha monoclonal antibodies were done, and argyrophilic nucleolar organizer regions (Ag-NOR) were counted to estimate the proliferative potential of 200 brain tumors. The findings were compared with the bromodeoxyuridine labeling index (BrdU LI), or S-phase fraction. The proliferating cell indexes (PCI), determined by Ki-67 and anti-DNA polymerase alpha monoclonal antibodies were higher and Ag-NOR more numerous in medulloblastomas, glioblastomas, and metastatic carcinomas than in astrocytomas or meningiomas. The Ki-67 and DNA polymerase alpha PCI correlated with the BrdU LI (r = 0.87 and r = 0.84, respectively) and with each other (r = 0.94). The number of Ag-NOR correlated less strongly with these indexes in some tumors. These findings suggest that Ki-67 and anti-DNA polymerase alpha monoclonal antibodies may be useful for estimating the proliferative potential of individual tumors in routine clinical practice. The number of Ag-NOR, however, does not always reflect the growth potential of each tumor. PMID- 8416717 TI - Role of early splenectomy in malignant lymphomas with prominent splenic involvement (primary lymphomas of the spleen). A study of 59 cases. AB - The outcomes were analyzed retrospectively of 59 cases of non-Hodgkin lymphoma (NHL) that included prominent splenic involvement (LPS). Forty-three patients had low-grade NHL, and 16 had intermediate or high-grade NHL. Forty of the 59 patients underwent splenectomy. Four patients died postoperatively before any treatment, and 10 others received no chemotherapy or radiation therapy. Twenty nine splenectomized and 16 patients whose spleens were not removed received chemotherapy or radiation therapy. One or more cytopenias were present in 45 patients (77%). Five (18%) of the 28 patients who initially were cytopenic underwent splenectomies that did not correct their blood disorders. The median actuarial survival was 108 months in splenectomized patients and 24 months in those not treated surgically (P = 0.0001). For the 40 splenectomized patients, a normal postoperative platelet count, an initial hemoglobin level of 110 g/l or more, and a postoperative hemoglobin level 110 g/l or more were associated with prolonged survival. These results suggest that cytopenias are frequent in LPS and that their reversal is observed after early splenectomy in 82% of cases. The absence of cytopenia after early splenectomy is associated with prolonged survival. PMID- 8416718 TI - Screening of esophageal and gastric cancer by occult blood bead detector. AB - A previously described bead method used to detect blood is simple, cheap, and effective; it can be repeated periodically. In 11 provinces in China, 233,825 persons (age range, 30-70 years) were screened. A positive occult blood test result was found in 28,557 persons (12%). Of these, 16,918 underwent a gastroscopy, resulting in the detection of 581 cancers that were located in similar frequencies in the esophagus, gastric cardia, and gastric body; 70% of the lesions were in an early or moderately advanced stage. Among 119 patients with early-stage cancer, the 3-year survival rate was 98.3%. After preliminary screening of cancer, the 4-year follow-up found a mortality rate in the group with negative occult blood test results that was only 25% of that of the positive group. Preliminary yearly screening would detect more early cancers and fewer advanced cancers. The mortality rate of esophageal and gastric cancer might be reduced drastically. PMID- 8416719 TI - Physician use among patients receiving cancer chemotherapy. AB - BACKGROUND: Little research has examined the volume and pattern of physician use or the scope of multiple physician use in patients with cancer. METHODS: The authors studied a sample of 259 patients with advanced cancer who received outpatient chemotherapy at two hospital clinics and eight private oncology practices. RESULTS: These patients reported regularly seeing an average of three different physicians an average of 15 times in 3 months. The number of physicians seen was strongly correlated with the number of reported visits (r = 0.65). Demographic and disease characteristics were associated only moderately with visit volume. Patients without a regular physician had a less concentrated pattern of visits to many doctors than did those with a regular doctor. CONCLUSIONS: This pattern of physician use among active treatment patients has not been described before and has implications for continuity of care. PMID- 8416720 TI - Dose-ranging antiemetic evaluation of the serotonin antagonist tropisetron in patients receiving anti-cancer chemotherapy. AB - BACKGROUND: Tropisetron (ICS 205-930) antagonizes the serotonin type 3 receptor and has antiemetic activity in animals given cisplatin. Its mean serum half-life in 11.1 hours. METHODS: In this dose-ranging trial, 22 patients undergoing anti cancer chemotherapy received 24 courses of a single intravenous infusion of tropisetron beginning 30 minutes before chemotherapy. Four dose levels were explored (range, 12-48 mg/m2). RESULTS: Toxicities were mild and included headache, transient elevations of serum alanine transaminase and/or aspartate transaminase levels, and sedation. No akathisia or acute dystonic reactions were observed. Thirty-six percent of patients had no emesis, and 58% had two or fewer emetic episodes. Ten patients received high-dose cisplatin (dose, > or = 100 mg/m2) as initial chemotherapy. Of these, 30% had no emesis, and 60% had two or fewer episodes. CONCLUSIONS: Tropisetron can be administered safely in the doses tested with no dose-limiting toxicities. The encouraging antiemetic efficacy, mild toxicities, lack of extrapyramidal effects, and convenience of a single 15 minute infusion regimen make this drug appropriate for study in additional trials. PMID- 8416721 TI - Cytokine levels in whole blood cell cultures as parameters of the cellular immunologic activity in patients with malignant melanoma and basal cell carcinoma. AB - For the determination of cellular immunity status, mitogen-induced lymphocyte proliferation tests are used, along with measurements of cytokine secretion. The authors have established a test system with whole blood cell cultures in which they measured the following cytokines: alpha-interleukin-1 (alpha-IL-1), interleukin-2 (IL-2), gamma-interferon (tau-IFN), and alpha-tumor necrosis factor (alpha-TNF) in the supernatants by enzymoimmunologic methods. With this system, the authors tested blood samples of 72 patients with malignant melanoma, 38 patients with basal cell carcinoma, and 315 healthy control subjects. In the blood cell cultures of the patients with melanoma, significantly lower values of the lymphokines tau-IFN and IL-2 were found, compared with those of the control subjects, and the levels of the monokines alpha-IL-1 and alpha-TNF were reduced. tau-IFN values correlated with different clinical stages. In contrast, the patients with basal cell carcinoma had equal values for all four cytokines as an age-matched control group. PMID- 8416722 TI - Differential expression of leukocyte differentiation antigens in small round blue cell sarcomas. AB - BACKGROUND: Small round blue cell sarcomas (SRBCS) comprise a group of cytomorphologically poorly differentiated neoplasms that are characterized by different histogenesis and biologic behavior. METHODS: Twenty-seven well characterized SRBCS were examined immunohistochemically to detect the expression of a panel of leukocyte differentiation (CD) antigens and class I (HLA-A,B,C) and class II (HLA-DR) major histocompatibility complex antigen. RESULTS: Although various cell surface antigens were detectable in SRBCS, the pan-leukocytic histiocytic CD53 antigen was absent in the neoplastic population of all tumors studied; this finding allowed the authors to discriminate these lesions from lymphomas and leukemias. Some antigens had a differential pattern of expression in the SRBCS group, in particular in the undifferentiated tumor cell populations. In most instances, neuroblastomas (NB) and ganglioneuroblastomas (GNB) were CD9+/CD24+/CD56+ but CD40-/HLA-A,B,C-. Rhabdomyosarcomas (RMS) were CD56+ in all specimens and CD9+ in many samples; generally, they showed CD24-/CD40-/HLA-A,B,C . Ewing sarcomas (ES) and peripheral primitive neuroectodermal tumors (pPNET) were HLA-A,B,C+/CD40+ but CD9-/CD24-/CD56- in most instances. Thus, with few exceptions, the expression of CD9 and CD56 and the simultaneous absence of HLA A,B,C and CD40 differentiated GNB, NB, and RMS from ES and pPNET. GNB, NB, and RMS differed in regard to their CD24 expression. CONCLUSIONS: These data show that various types of SRBCS have different patterns of cell surface antigens. Therefore, these antigens are considered to be helpful in the immunophenotypic subclassification of SRBCS: The immunophenotypic similarities between ES and pPNET, however, might be an additional argument for a close relationship between these two lesions. PMID- 8416723 TI - Aggressive rectal lymphoma of large granular lymphocytes with the histologic feature of an angiocentric growth pattern. AB - The authors report an unusual large cell lymphoma of the rectum composed of large granular lymphocytes (LGL) with histologic characteristics of an angiocentric growth pattern. Immunophenotyping showed that most of the tumor cells were CD3-, CD4-, CD8+, CD16+, CD56+, and CD57-. Fine structural analysis of the tumor cells found substantial numbers of electron-dense granules. Genotypic investigation showed a germline configuration of the T-cell receptor beta and gamma chain genes and the immunoglobulin heavy chain gene. The clinical course was aggressive, with rapid dissemination to the lungs, liver, and subcutis. The lesion was resistant to chemotherapy. There was, however, no evidence for peripheral blood or bone marrow involvement. This case report demonstrates the need for continued inquiry into the possible association of LGL with angiocentric lymphoproliferative lesions and gut-associated T-lymphocyte lesions. PMID- 8416724 TI - Severe aplastic anemia preceding acute lymphoblastic leukemia. AB - BACKGROUND: Bone marrow aplasia preceding acute lymphoblastic leukemia (ALL) is a rare condition that usually affects children. The ALL generally follows the recovery of normal blood counts and most commonly occurs within 6 months of the onset of aplasia. The case of a patient with severe aplastic anemia is reported in whom ALL developed 15 months after the initial diagnosis of aplastic anemia. A literature search found 23 cases of ALL after a period of aplasia or hypoplasia. This patient's disease, however, was different from all previously reported ones. The severe aplasia lasted 15 months before being followed by ALL. There was no recovery of blood counts before the onset of ALL. METHODS: A review of the literature found 23 case reports in which aplasia or hypoplasia preceded ALL; these patients also had pancytopenia of the peripheral blood. Excluded from this review were patients whose bone marrow was hypoplastic, but who did not have pancytopenia because these did not have "aplastic anemia" as their initial disease. RESULTS: Analysis of the reported patients showed that most were girls 10 years of age or younger. There was an overwhelming prevalence of fever, which in several instances, might have had an infectious cause. ALL most commonly occurred within 6 months of the aplasia and usually followed the recovery of normal blood counts. CONCLUSIONS: Patients with ALL after a prolonged period of aplasia have several common characteristics including female sex, young age, and the prevalence of fever, often associated with an infectious illness. ALL usually follows the recovery of blood counts and occurs within 6 months of the onset of aplasia. The pathophysiology of this patient's disease(s) is still unclear. He could have had two unrelated disorders or a two-step leukemic process that followed a stem cell "insult." This patient had an antecedent hepatitis A infection 3 months before aplasia occurred. However, the authors were unable to identify with certainty any other event that might have caused additional bone marrow injury. PMID- 8416725 TI - Fluorescence detection of tumors. Early diagnosis of microscopic lesions in preclinical studies. AB - BACKGROUND: The growth of microscopic tumor lesions at or beyond treatment field margins poses a major problem in the diagnosis and treatment of cancer. Early detection techniques that clearly define the location or field spread of disease may improve the planning of disease treatment. METHODS: In vivo fluorescence photometry is a non-imaging technique that digitally displays relative fluorescence values in volts proportional to the luminescence intensity detected by a silicon photodiode. The sensitivity of the instrument has allowed the detection of micrometastases in preclinical studies. RESULTS: Statistical analysis demonstrates that the photosensitizer Photofrin (dihematoporphyrin ether and/or ester) (Quadra Logic Technologies, QLT, Vancouver, B.C., Canada), currently used for photodynamic therapy, administered in doses lower than those used in clinical studies, is useful for the detection of occult disease. With the drug doses used, cutaneous photosensitivity was avoided in the animal models tested. The results in Lobund-Wistar rats with transplantable prostatic adenocarcinoma (PA-III) demonstrated the utility of this technique in detecting clinically occult disease, with a prediction rate of approximately 94% with drug doses as low as 0.25-0.5 mg/kg. CONCLUSIONS: With the use of the hamster buccal cavity model involving the initiation and promotion of premalignant and malignant conditions by 9,10 dimethyl-1,2-benzanthracene, the technique could discern these two stages of disease with significance levels that were less than 0.05 and 0.01, respectively. PMID- 8416726 TI - Conservation therapy for invasive lobular carcinoma of the breast. PMID- 8416727 TI - Comparison of esophagocardiac and more distal gastric cancer in patients with prior ulcer surgery. AB - BACKGROUND: Adenocarcinoma of the gastric cardia and/or lower esophagus differs from cancer of the more distal stomach. METHODS: The authors compared the proportion of patients with these two types of cancer who underwent surgery for peptic ulcers more than 5 years before the diagnosis of cancer during the periods 1977-82 and 1983-88. RESULTS: The total number of patients with esophagocardiac cancer increased from 99-241 between the two periods, but the number with previous ulcer surgery remained the same (n = 7) in each period. The total number of more distal gastric cancers decreased from 262 to 237 between the two periods, but the number with previous ulcer surgery increased from 9-26 (P = 0.002). Only 4 of 14 patients with esophagocardiac cancer and a previous ulcer had a partial gastrectomy compared with 27 of 35 patients with more distal gastric cancers (P = 0.005). CONCLUSION: These data indicate a relationship between partial gastrectomy and the late development of gastric cancer, but this finding is confined to cancers not involving the cardia. PMID- 8416728 TI - Recurrent acromegaly resulting from ectopic growth hormone gene expression by a metastatic pancreatic tumor. AB - BACKGROUND: Acromegaly is usually the result of a pituitary growth hormone (GH) cell adenoma or is more rarely due to ectopic secretion of GH-releasing hormone (GHRH). The authors previously described a more unusual form of acromegaly secondary to ectopic GH synthesis by a pancreatic islet cell tumor. METHODS: One year after tumor resection and transient disease remission, multiple abdominal metastases were identified with accompanying elevated levels of circulating GH and insulin-like growth factor-1 (IGF-1). Serial 24-hour GH sampling was performed before and after intravenous GHRH or thyrotropin releasing hormone (TRH) administration during treatment with bromocriptine; treatment with the somatostatin (SRIF) analogue octreotide; or no treatment. RNA from abdominal tumor tissue was extracted and subjected to Northern gel electrophoresis and GH hybridization analysis. RESULTS: Neither GHRH nor TRH resulted in stimulation of the elevated GH levels. Bromocriptine and octreotide did not suppress GH secretion but attenuated the thyroid stimulating hormone (TSH) response to TRH administration. Octreotide (as much as 1500 micrograms/d) was clinically, biochemically, and radiographically ineffective. GH-secreting abdominal tumor tissue expressed a 0.9-kb mRNA transcript consistent with the size of authentic human GH mRNA. CONCLUSION: The natural history and ectopic nature of a GH producing pancreatic carcinoma has been documented, with biochemical remission occurring after initial tumor resection, with autonomous GH hypersecretion after tumor recurrence, and with RNA analysis demonstrating ectopic activation of GH gene transcription. PMID- 8416729 TI - Expression of blood group-related antigens in normal and malignant pancreatic tissue correlated with genotype of the patient defined by saliva glycoprotein. AB - BACKGROUND: Seven murine monoclonal antibodies (MoAb) detecting blood group antigens of the Lewis system and their sialylated derivatives were used to define their immunohistochemical distribution in normal and malignant pancreatic tissues. The specific blood group antigens studied included Lewis(a) (Le(a)), Lewisb (Leb), Lewisx (Le(x)), Lewis(y) (Le(y)), sialyl-Lewis(a) (s-Le(a)), sialyl lacto-N-tetraose (s-LNT), and sialyl-Lewisx (s-Le(x)). METHODS: The expression of these antigens was analyzed by immunoperoxidase technique in pancreatic tissue of patients with (n = 27) and without (n = 19) pancreatic cancer. The genetic background of their secretor status and their Lewis phenotypes were determined by the enzyme-linked immunosorbent assay using purified salivary glycoprotein and MoAb against eight different blood group-related antigens (Le(a), Leb, Le(x), Le(y), H1, H2, A, B2), and the putative genotypes of the patients were classified as follows: SeLe, Sele, seLe, and sele. RESULTS: The following results were obtained: (1) In normal pancreas, Le(a), s-Le(a), s-LNT, and s-Le(x) were expressed in ducts, but their expression was restricted to the luminal side of the cytoplasm (cytoplasmic type with positive polarity). Leb and Le(y) showed similar patterns of expression in ducts and some parts of the acinus. In contrast, Le(x) was absent in most cases. (2) Expression of Leb was observed in normal pancreatic ducts in all 46 patients, regardless of their genotypes. Le(a) and s-Le(a) were not expressed in specimens from patients with the seLe and sele saliva phenotypes. (3) In pancreatic cancer, the following antigens were expressed cytoplasmically in the proportion of cases indicated: Le(a) (78%), Leb (85%), Le(x) (33%), Le(y) (44%), s-Le(a) (78%), s-LNT (89%), and s-Le(x) (85%). They also were detected in the surrounding stroma. This pattern of expression is distinct from that found in normal pancreatic tissue. Patients with Sele and sele genotypes did not express Le(a) or s-Le(a), except in one case. (4) Serum levels of carbohydrate antigen 19-9 (CA 19-9) were not elevated in patients with pancreatic cancer with the Sele and sele genotypes but were elevated in most patients with SeLe and seLe genotypes. CONCLUSION: The overall findings indicate that Lewis-related antigens act as pancreatic tissue-related antigens, depending in part on salivary phenotypes of the patient. With proper antigen selection and the determination of secretor status, these anti-blood group MoAb may be of clinical utility in the diagnosis of pancreatic cancer. PMID- 8416730 TI - Papillary cystic tumors of the pancreas. Assessment of their malignant potential. AB - BACKGROUND: Although the biologic characteristics of papillary cystic tumors (PCT) generally indicate a good prognosis, a malignant form has been reported. METHODS: Twenty-two examples of PCT were examined to assess their malignant potential by histologic, flow cytometric, and immunohistochemical studies. RESULTS: Three had hepatic, peritoneal, and/or lymph node metastases (metastasizing tumors [MT]); the other 19 PCT had no such metastatic features (nonmetastasizing tumors [NMT]). Venous invasion was detected in all three MT and two NMT. Compared with the 19 NMT, the MT had a higher nuclear grade and more prominent necrobiotic nests characterized by aggregates of cells with pyknotic nuclei and eosinophilic cytoplasm. In the flow cytometric analysis of cellular DNA content, one MT was aneuploid, and eight NMT were diploid. Immunohistochemically, there was no difference between the MT and NMT. CONCLUSIONS: These results support the assumption that venous invasion, nuclear grade, and prominent necrobiotic nests are useful as histologic parameters to detect the malignant potential of PCT. PMID- 8416731 TI - Recent advances in the treatment of advanced colorectal cancer. AB - The treatment of advanced colorectal cancer has improved in recent years. Prospective randomized trials comparing innovative therapies with the "standard" bolus dose of 5-fluorouracil (5-FU) found increased response rates after biochemical modification of the drug, infusion administration of 5-FU, and direct intrahepatic arterial infusion. Although the impact on survival of these techniques has been minimal, it is possible that these innovative approaches provide an incremental survival advantage for certain subgroups of patients that may be the foundation for additional therapeutic improvements in the future. PMID- 8416732 TI - Cardiac tamponade caused by primary lung cancer and the management of pericardial effusion. AB - BACKGROUND AND METHODS: Between 1978 and 1990, 51 cases of pericardial effusion secondary to lung cancer were treated at the National Cancer Center Hospital by creating a pericardial window, using the subxiphoid approach, that was connected to a water-sealed drainage system. RESULTS: Most patients had advanced disease, such as distant metastasis (76%), pleural effusion (88%), and clinical Stage N2 or N3 disease (98%). Forty-five patients had cardiac tamponade, and six had no symptoms attributable to pericardial effusion. Cardiac tamponade was the initial manifestation of lung cancer in only 3 patients; it was a late manifestation in 48. Of those specimens that were examined cytologically, 92% had positive findings. The interval from creation of the pericardial window until removal of the drainage tube ranged from 4-135 days (median, 11 days). The interval was significantly longer in patients who previously had received thoracic radiation therapy (P < 0.05). The overall median survival was 80 days, and the 1-year survival rate was 10.5%. Postmortem examination showed that constrictive heart failure caused by pericardial lesions was the major contributory cause of death in 32% of patients. Using multivariate analysis, factors indicating a poor prognosis were: (1) the interval from the diagnosis of lung cancer to pericardial effusion development (P = 0.005) and (2) the absence of prior surgery (P = 0.007). CONCLUSIONS: The creation of a pericardial window effectively treated pericardial effusion in 85% of cases. However, the role of intrapericardial instillation of anticancer or sclerosing agents was unclear in this retrospective analysis. PMID- 8416733 TI - Development and characterization of an isolated and perfused tumor and skin preparation for evaluation of drug disposition. AB - The purpose of this study was to develop a model in which the regional pharmacokinetics of a drug in tumor and nontumorous tissue could be evaluated under a variety of physiological conditions. To this effect, the growth of a human choriocarcinoma cell line (JAR) was evaluated in pigs immunosuppressed with 25 mg cyclosporine/kg every 24 h. During an initial study, we demonstrated that suspensions containing approximately 3 million JAR cells with and without 1 million normal human fibroblasts injected s.c. into the inguinal region of pigs resulted in the growth of tumors consisting primarily of polygonal neoplastic cells. Multinucleate tumor cells, inflammatory cells, necrotic debris, and vascular endothelial cells were also present. Maximal tumor size was noted on day 12, after which time tumor regression occurred. The coinoculation of fibroblasts resulted in significantly larger tumors. Two single pedicle, axial pattern tubed flaps were created in the inguinal area of 4 pigs. JAR cells and fibroblasts were transplanted to one flap to allow for tumor formation. The other flap served as a nontumorous control. Both flaps were removed for perfusion with a physiological solution 11 days later. Glucose utilization, lactate concentrations, lactate dehydrogenase activities, and microscopic evaluation of skin samples were used to assess flap viability. All flaps remained viable for 8 h of perfusion. The only differences detected between nontumorous and tumor flaps was the initial perfusion pressure which was significantly lower in tumor flaps (P < 0.05). The isolated perfused tumor and skin flap is unique in that it consists of a tumor surrounded by normal tissue with an intact microvascular system and can be utilized to design regional pharmacokinetic studies describing drug distribution in tumor tissue. PMID- 8416734 TI - Modulation by adjuvants and carriers of the immunogenicity in xenogeneic hosts of mouse anti-idiotypic monoclonal antibody MK2-23, an internal image of human high molecular weight-melanoma associated antigen. AB - The mouse anti-idiotypic monoclonal antibody (mAb) MK2-23 recognizes an idiotope in the antigen-combining site of the immunizing anti-human high-molecular-weight melanoma-associated antigen (HMW-MAA) mAb 763.74. Administration with an adjuvant of mAb MK2-23 conjugated to a carrier has been shown to induce anti-HMW-MAA antibodies both in syngeneic hosts and in patients with malignant melanoma. Adjuvant and carrier are required for the induction of anti-HMW-MAA immunity in BALB/c mice immunized with mAb MK2-23. Whether both adjuvant and carrier are required also in patients with malignant melanoma is not known and cannot be deduced from results obtained in a syngeneic animal model system. Therefore the present study has evaluated for the first time the effect of a carrier and an adjuvant on the immunogenicity of mAb MK2-23 in a xenogeneic host. Rabbits were selected for this purpose, since they have a constitutive expression of HMW-MAA in their normal tissues with a distribution similar to that in humans. The combined use of an adjuvant and a carrier enhances the immunogenicity of mAb MK2 23 in rabbits markedly more than each of them individually. Specifically, all the rabbits immunized with mAb MK2-23 conjugated to keyhole limpet hemocyanin (KLH) and mixed with Freund's adjuvant (FA) produced antibodies which were shown with serological and immunochemical assays to be specific for HMW-MAA and to be both IgG and IgM. In contrast anti-HMW-MAA antibodies were detected in only one of the 3 rabbits immunized with mAb MK2-23 mixed with FA and were not detected in the rabbits immunized with mAb MK2-23 conjugated to KLH or with mAb MK2-23 without KLH and FA. These results indicate that active specific immunotherapy with mAb MK2-23 in patients with malignant melanoma is likely to benefit from the use of a carrier and an adjuvant, provide additional evidence that mAb MK2-23 bears the internal image of HMW-MAA, and suggest that the immune response elicited by mAb MK2-23 is T-cell dependent. PMID- 8416735 TI - Oxidative injury rapidly activates the heat shock transcription factor but fails to increase levels of heat shock proteins. AB - When cells are exposed to heat shock, heavy metals, amino acid analogues, and other stresses, the heat shock transcription factor (HSF) is activated. The HSF then binds to the promoter of the heat shock genes, stimulating transcription of the heat shock proteins. Here, we demonstrate that exposure of NIH-3T3 cells to oxidants (H2O2 or menadione) also causes activation of the HSF. This activation is not blocked by inhibitors of protein synthesis (cycloheximide) or by inhibitors of protein kinases (2-aminopurine or genistein). In addition, the oxidant activated HSF is located in the nucleus of the cells. However, oxidant activation of the HSF does not result in the accumulation of hsp70 mRNA or of heat shock proteins. This is in contrast to the accumulation of heat shock proteins seen after heat shock activation of the HSF. This suggests that oxidant induced activation of HSF binding may have a function different from that of heat induced activation of HSF binding. PMID- 8416736 TI - Lactotetraose series ganglioside 3',6'-isoLD1 in tumors of central nervous and other systems in vitro and in vivo. AB - Two monoclonal antibodies, DMAb-21 and DMAb-22, directed against the lactotetraose series ganglioside-associated epitope IV3NeuAc,III6-NeuAcLcOse4Cer (3',6'-isoLD1), were found to define the minimum binding epitope NeuAc(or NeuGc)alpha 2-3Gal beta 1-3(NeuAc or NeuGc)alpha 2-6GlcNAc. The distribution of 3',6'-isoLD1 in cultured cell lines and derived xenografts of primary tumors of the human central nervous system and of embryonal or neuroectodermal tumor derivation was determined. Only 4 of 26 cell lines, 3 teratomas and 1 pancreatic adenocarcinoma, expressed detectable 3',6'-isoLD1 when cultured in vitro; none of 14 tested glioma lines, including 2 that expressed the monosialo-precursor IV3NeuAcLcOse4Cer in vitro, expressed detectable levels. Expression of 3',6' isoLD1 was more frequent when neoplastic cells were grown in xenograft form in athymic mice; 4 of 10 glioma and 2 of 2 teratoma xenograft ganglioside extracts were positive for 3',6'-isoLD1. The absence of 3',6'-isoLD1 in cultured tumor cells of the central nervous system and its proportional increased presence in tumor cells of the same origin grown in vivo further supports previous studies suggesting that ganglioside expression may be modified by environmental forces. The expression of lacto series gangliosides both in vitro and in vivo by teratoma and pancreatic adenocarcinoma cells, as opposed to only in vivo expression by glioma cells, suggests that tissue-specific forces may also exist. Immunohistochemical localization of 3',6'-isoLD1 in frozen sections of primary central nervous system neoplasms including those of glial and nonglial origin was performed; 20 of 30 (67%) of glial tumors were positive. Among nonglial tumors, 21 of 34 (62%) of epithelial cancers were reactive with anti-3',6'-isoLD1 monoclonal antibodies; notably negative were carcinomas of the ovary and lung carcinomas of all subtypes. Lymphomas and infiltrative lymphocytes were uniformly negative. The restriction of 3',6'-isoLD1 expression within the human central nervous system to periods of fetal-neonatal astroglial proliferation, to intense reactive astrocytosis, and to primary neoplasms, and the production of specific monoclonal antibodies to this epitope provide a specific complex for immunolocalization and, eventually, immunotherapy. PMID- 8416737 TI - Theophylline induced differentiation provides direct evidence for the deregulation of c-myc in Burkitt's lymphoma and suggests participation of immunoglobulin enhancer sequences. AB - Most of the evidence that supports the hypothesis that the c-myc gene is abnormally regulated in Burkitt's lymphoma (BL) is indirect. The putative abnormal expression of c-myc is likely, at least in part, to be a consequence of the usurpation of its regulatory sequences by immunoglobulin enhancer elements, which are invariably juxtaposed to c-myc by the translocations associated with this tumor (C. M. Croce, J. Erikson, A. Ar-Rushdi, D. Aden, and K. Nishikura, Proc. Natl. Acad. Sci. USA, 81: 3170-3174, 1984). We have developed a differentiation induction model system to examine this issue more directly. In a variety of non-BL cell lines, differentiation induction results in the down regulation of c-myc (G. P. Studzinski, A. K. Bhandal, and Z. S. Brelvi, Proc. Soc. Exp. Biol. Med., 179: 288-295, 1985; Y. Matsui, R. Takahasi, K. Minara, T. Nakagawa, T. Koizumi, Y. Nakao, T. Sugiyama, and T. Fugita, Cancer Res., 49: 1366 1371, 1985; T. Mitchell, E. Sariban, and D. Kufe, Mol. Pharmacol., 30: 398-402, 1986; Z. S. Brelvi, and G. P. Studzinski, J. Cell. Physiol., 128: 171-179, 1986). Since BL is of B-cell origin, differentiation is associated with persistent or increased expression of immunoglobulin genes. Therefore, if c-myc and c-mu are coregulated in BL via immunoglobulin enhancer sequences, persistent or increased expression of the c-myc gene, rather than down-regulation, should occur in differentiated BL cells. Differentiation was induced in four BL cell lines with theophylline (7 x 10(-3) M), and mRNA was examined by Northern blot analysis. In all four BL lines studied (JD38, AG876, KK124, and Daudi), there was persistent or increased expression of both c-mu and c-myc genes (detected with a third exon c-myc probe), in contrast to the decreased expression of the c-myc gene observed in the three Epstein-Barr virus transformed lines studied (A3317, TC84, and CB34). In the BL cell line, JD38, the c-myc gene is truncated (the second and third exons are translocated to chromosome 14 while the first exon remains on chromosome 8). In this line, we demonstrated that theophylline induced differentiation results in down-regulation of the first exon while the level of expression of the translocated second and third exons remains unchanged or increases.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8416738 TI - M(r) 92,000 type IV collagenase is increased in plasma of patients with colon cancer and breast cancer. AB - Overproduction of matrix metalloproteinases (MMPs) is a common characteristic of metastatic cancer cells. Since MMPs can be identified in plasma, we proposed that enhanced MMP-9 secretion by invasive cancer cells may be detected by plasma assay. To this end, we developed a specific sandwich enzyme-linked immunosorbent assay which uses two mouse monoclonal antibodies to human M(r) 92,000 type IV collagenase (MMP-9). The plasma concentration of MMP-9 (mean +/- SD) in 60 healthy subjects (9 +/- 11 ng/ml), 136 patients without cancer, and 179 patients with cancer of the lung, genitourinary tract, or lymphomas-leukemias did not differ significantly. In contrast, plasma MMP-9 was significantly increased (P < 0.01) in 122 patients with gastrointestinal tract cancer and breast cancer (18 +/ 23 and 21 +/- 22 ng/ml, respectively). Whereas carcinoembryonic antigen levels were significantly increased in patients with stage IV gastrointestinal cancer, MMP-9 concentrations were not significantly increased in patients with metastatic disease as compared to those with nonmetastatic cancer. Combining both assays improves sensitivity of detection of colon cancer. MMP-9 was also significantly increased during pregnancy which is consistent with the extensive ongoing tissue remodeling and the leaching of the tissue proteinase into plasma. PMID- 8416739 TI - Tumor-secreted vascular permeability factor/vascular endothelial growth factor influences photosensitizer uptake. AB - The role of vascular permeability in the preferential accumulation of photosensitizers in tumor tissue was investigated. Two murine tumors [experimental mammary tumor carcinoma (EMT-6) and methylcholanthrene-induced rhabdomyosarcoma (M1S)] and a human bladder carcinoma (EJ) were grown s.c. on the flank in athymic nude mice and analyzed for in vivo vessel permeability, vascular permeability factor (VPF) secretion, and accumulation of the photosensitizer, chloroaluminum sulfonated phthalocyanine. In vivo tumor vessel permeability and vascular volume were quantitated by measuring Evans blue extravasation and accumulation of a high molecular weight fluoresceinated dextran, respectively. VPF was isolated from serum-free tumor cell conditioned medium using heparin Sepharose affinity chromatography. Dot and Western blots stained with anti-VPF antiserum positively identified VPF in samples from each tumor. Chloroaluminum sulfonated phthalocyanine pharmacokinetics in tumor-bearing mice were measured using a fiber-based spectrofluorometer. In vivo vessel permeability was found to be greatest in M1S tumors, next in EMT-6 tumors and finally in EJ tumors. Consistent with in vivo data, M1S and EMT-6 tumor cells in culture secrete significantly more VPF than EJ tumor cells. Chloroaluminum sulfonated phthalocyanine accumulation was approximately 2 times greater in M1S and EMT-6 tumors compared to EJ tumors. Our data present evidence that photosensitizer accumulation can be correlated to in vivo tumor vessel permeability and VPF secretion of that tumor. Taken together, the data support the hypothesis that vascular permeability differences among tumors play a significant role in the uptake and retention of photodynamic agents. PMID- 8416740 TI - Overexpression of basic fibroblast growth factor complementary DNA in Ha-ras transformed cells correlates with a decreased incidence of tumor necrosis. AB - Solid tumors often contain poorly vascularized necrotic regions. In order to determine how such regions are formed within tumors and to identify substances which affect their formation, we have transformed nontumorigenic BALB/c 3T3 cells with an activated Ha-ras oncogene. Cells that were derived from independent clones of Ha-ras-transformed cells were injected s.c. into BALB/c mice. When the resulting tumors reached a weight of about 4 g, the mice received i.v. injections of 51Cr-labeled RBC. The distribution of the labeled RBC in various areas within the tumors was determined. The peripheral parts of these tumors contained viable cells, numerous blood vessels, and high concentrations of labeled RBC. The cores of the tumors on the other hand appeared necrotic, accumulated much lower concentrations of labeled RBC, and contained largely fibrous material and almost no viable cells. An expression vector containing the complementary DNA of human basic fibroblast growth factor was stably transfected into cells derived from two of the Ha-ras-transformed clones. Transfected clones of cells which produced low or intermediate amounts of basic fibroblast growth factor developed, following their injection into BALB/c mice, into tumors resembling the tumors that develop from the parental Ha-ras-transformed cells. In contrast, about one-half of the clones which produced large amounts of basic fibroblast growth factor developed into tumors which were composed almost totally of live tissue and were almost completely devoid of necrotic areas. In these tumors the labeled RBC were distributed evenly throughout the tumor. PMID- 8416741 TI - MDM2 gene amplification in metastatic osteosarcoma. AB - The human homologue of the murine double minute 2 gene (MDM2), a p53-binding protein which may act as a regulator of p53 protein function, has recently been cloned. Initial studies of this gene in a variety of human tumors have shown frequent gene amplification in most types of sarcomas, including osteosarcomas. Amplification of the MDM2 gene may produce a functional inactivation of the p53 protein. To examine possible clinical or pathological correlates of MDM2 gene amplification in osteosarcoma, we studied 28 specimens on 26 patients with high grade osteosarcoma (16 primary, 11 metastatic, and 1 local recurrence) for MDM2 gene amplification by Southern blot analysis, using two MDM2 complementary DNA probes isolated by polymerase chain reaction. Four specimens (14%) showed amplification, including 3 metastases and 1 local recurrence. None of the primary osteosarcoma specimens had detectable MDM2 gene amplification. None of the specimens tested showed MDM2 gene rearrangement. In the present series, MDM2 gene amplification was detected significantly more frequently in metastatic or recurrent osteosarcomas than it was in primary osteosarcomas (P = 0.02). Our data suggest that MDM2 gene amplification may be associated with tumor progression and metastasis in osteosarcoma. Further investigation is warranted on the potential clinicopathological correlates of MDM2 gene amplification in osteosarcoma. PMID- 8416742 TI - Expression of oligodendrocyte-associated genes in cell lines derived from human gliomas and neuroblastomas. AB - Two putative human oligodendroglioma cell lines were examined for the expression of the oligodendrocyte-associated genes, 2',3'-cyclic nucleotide-3' phosphodiesterase, myelin basic protein, myelin proteolipid proteins, and myelin associated glycoprotein. The expression of these genes also was examined in control astrocytoma and neuroblastoma cell lines. In addition, the expression of the non-oligodendrocyte-specific genes, glial fibrillary acidic protein (GFAP), neuron-specific enolase and neurofilaments (NF) NF-L and NF-M also were examined. All the cell lines expressed 2',3'-cyclic nucleotide 3'-phosphodiesterase, neuron specific enolase, and vimentin, and none expressed myelin-associated glycoprotein. The "oligodendrocyte-specific" myelin proteolipid protein mRNAs and the "neuron-specific" NF-L mRNA were expressed in the two astrocytoma cell lines, which also expressed GFAP. Expression of intermediate filament protein genes was more restricted. The astrocytoma, neuroblastoma, and oligodendroglioma cell lines expressed only GFAP, NF-M, and cytokeratin K7, respectively. These results: (a) provide molecular data confirming the classification of the two cell lines as oligodendrogliomal and suggest that their molecular profiles are indicative of immature oligodendrocytes; (b) demonstrate the expression of cytokeratins in oligodendrogliomal cell lines and suggest that apparent GFAP expression in oligodendrogliomas detected by immunocytochemical methods may be due to cross reactivity with cytokeratins, with which they share common polypeptide sequence; and (c) indicate that astrocytoma cell lines can exhibit a "mixed" phenotype, expressing genes associated with fully differentiated oligodendrocytes and neurons. PMID- 8416743 TI - Exploiting the lacZ reporter gene for quantitative analysis of disseminated tumor growth within the brain: use of the lacZ gene product as a tumor antigen, for evaluation of antigenic modulation, and to facilitate image analysis of tumor growth in situ. AB - We extend use of the lacZ reporter gene for tumor biology. Intracerebral growth of 9L/lacZ, a gliosarcoma cell line that stably expresses lacZ, was evaluated in syngeneic rats. The reporter gene product, Escherichia coli-derived beta galactosidase (beta-gal), was detected histochemically on tissue sections. This permits visualization of disseminated tumor and, as shown here, facilitates image analysis. We show that the beta-gal marker protein itself can serve as a tumor antigen in appropriate contexts. Quantitative image analysis of tumor areas is used to show that immunization with beta-gal protects against tumor growth. Abnormal beta-gal- areas are easily detected, facilitating study of antigenic modulation. The tumor studied did not escape through this mechanism. All abnormal beta-gal- areas examined were shown to reflect accumulation of inflammatory or reactive cells, not tumor. Taken together, these findings show several ways in which the lacZ reporter gene can be exploited to facilitate quantitative analysis of disseminated tumor growth within the brain. They draw attention to the growing appreciation that tumor antigens need not be cell surface molecules. PMID- 8416744 TI - Immunoelectron microscopic localization of transforming growth factor beta 1 and latent transforming growth factor beta 1 binding protein in human gastrointestinal carcinomas: qualitative difference between cancer cells and stromal cells. AB - Transforming growth factor beta 1 (TGF-beta 1) is secreted as an inactive complex associated with latent TGF-beta 1 binding protein (LTBP). Tissue localization of these proteins has not been fully understood in human pathological conditions. We examined the immunohistochemical localization of TGF-beta 1 precursor (proTGF beta 1) and LTBP in carcinomas and granulation tissue in the human gastrointestinal tract at the light and electron microscopic levels. In normal tissue, endothelial cells and granulocytes sporadically showed immunoreactivity for proTGF-beta 1, while epithelial cells were all negative. In cancer tissue, both cancer cells and stromal cells (fibroblasts, macrophages, and endothelial cells) were positive for proTGF-beta 1, more frequently in diffuse-type gastric carcinomas than in differentiated-type adenocarcinomas. Immunoelectron microscopy revealed that proTGF-beta 1 was localized in rough endoplasmic reticulum and perinuclear cisternae in fibroblasts, macrophages, and endothelial cells in cancer stroma and in fibrous granulation tissue. In contrast, the intracellular localization of proTGF-beta 1 in carcinoma cells was predominantly observed in the cytosol (cytoplasmic matrix). This finding suggests disarranged or blocked intracellular transportation of proTGF-beta 1 in cancer cells. The immunoreactivity for LTBP was not observed in the normal epithelial cells. It was localized in cancer stroma, not in cancer cells. Ultrastructurally, LTBP was located in the extracellular matrix around fibroblasts and smooth muscle cells. The intracellular immunoreactivity for LTBP was observed in rough endoplasmic reticulum of fibroblasts and smooth muscle cells, the same as in granulation tissue. These results suggest that gastrointestinal carcinoma cells produce no or a small amount of LTBP in vivo. Our investigation suggests that extensive fibrosis in both cancer stroma and granulation tissues may be promoted by TGF beta 1 mainly secreted from stromal cells. PMID- 8416745 TI - 2,2'-Bipyridyl-6-carbothioamide and its ferrous complex: their in vitro antitumoral activity related to the inhibition of ribonucleotide reductase R2 subunit. AB - 2,2'-Bipyridyl-6-carbothioamide (BPYTA), a synthetic compound with antitumoral activity, is characterized by chelating properties because of the N*-N*-S* tridentate ligand system and is therefore comparable to alpha-(N)-heterocyclic carboxaldehyde thiosemicarbazones which are potent inhibitors of ribonucleotide reductase (RR). Electron paramagnetic resonance studies on the small subunit of mouse recombinant RR (R2) demonstrated that BPYTA can destroy the R2 tyrosyl radical only if Fe(II) is present (73% destruction at 50 microM, after 20 min of contact). The R2 inhibition was reversible and time dependent. Studies on tumoral lines confirmed that the main cell target of BPYTA is RR and demonstrated that the iron-complexed form compared to the nonchelated form has some difficulty in crossing the cell membrane. Spectrophotometric and electron paramagnetic resonance studies clearly indicated that BPYTA chelates iron only when this is reduced and that the BPYTA-Fe(II) complex is stable in the presence of oxygen. From reported results we conclude that BPYTA is a powerful RR inhibitor (R2 subunit) which has a different mechanism of action from that of Desferal. It has some properties in common with alpha-(N)-heterocyclic carboxaldehyde thiosemicarbazones, but they are not identical. It would be interesting to do further studies on the BPYTA mechanism of action and evaluate the in vivo antitumoral activity of the preformed complex. PMID- 8416746 TI - Inhibition of skin tumor promotion by restriction of fat and carbohydrate calories in SENCAR mice. AB - The purpose of this research was to compare the influence of calorie restriction by removal of fat with calorie restriction by removal of carbohydrate on the promotion of skin tumorigenesis in mice. Female SENCAR mice were initiated with 7,12-dimethylbenz(a)anthracene (10 nmol, single topical treatment) and fed calorie-restricted diets during and following promotion with 12-O tetradecanoylphorbol-13-acetate (2 micrograms, topically, twice a week for 20 weeks). Control diet (American Institute of Nutrition-based formulation) was compared with a diet in which calories from fat and calories from carbohydrate were similar [balanced high fat (BHF)] in ad libitum-fed groups. Restricted animals were fed diets such that 35% of the calories from fat [high carbohydrate, calories restricted from fat (HCR)] or from carbohydrate (high fat, calories restricted from carbohydrate) were restricted, but other intake was equivalent to the BHF group. Results showed an inhibition of papilloma number in both restricted groups and the inhibition was greatest in the HCR mice. Larger papillomas were observed on mice in the control, BHF, and high-fat, calories restricted from carbohydrate diet groups than on mice in the HCR group. The pattern of carcinoma development was similar in the mice in the freely fed control and BHF groups. Restriction of calories from either fat or carbohydrate delayed the rate and reduced the incidence of carcinoma development. Carcinoma incidence did not differ between mice fed the high-fat, calories restricted from carbohydrate and HCR diets. PMID- 8416747 TI - Effects of sodium nitrite and catechol, 3-methoxycatechol, or butylated hydroxyanisole in combination in a rat multiorgan carcinogenesis model. AB - Effects of simultaneous treatment with NaNO2 and butylated hydroxyanisole, catechol, or 3-methoxycatechol were examined in a rat multiorgan carcinogenesis model. Groups of 15 animals were given a single i.p. injection of 100 mg/kg of body weight diethylnitrosamine, 4 i.p. injections of 20 mg/kg of body weight N methylnitrosourea, 4 s.c. injections of 40 mg/kg of body weight dimethylhydrazine, p.o. treatment with 0.05% N-butyl-N-(4 hydroxybutyl)nitrosamine in the drinking water for the first 2 weeks and p.o. treatment with 0.1% 2,2'-dihydroxy-di-n-propylnitrosamine in the drinking water for the next 2 weeks of the initial 4-week initiation period. Starting 3 days after the completion of these carcinogen treatments, animals were given diets containing 2% butylated hydroxyanisole, 0.8% catechol, 2% 3-methoxycatechol, or basal diet either alone or in combination with 0.3% sodium nitrite until week 28, when complete autopsy was performed. Histological examination showed that NaNO2 strongly enhanced development of forestomach lesions but inhibited that of glandular stomach lesions in rats simultaneously given catechol or 3 methoxycatechol with or without prior carcinogen exposure. 3-Methoxycatechol promoted esophageal carcinogenesis either with or without NaNO2, but promoting effects of catechol were evident only in the presence of NaNO2. In addition, treatment with NaNO2 after carcinogen exposure enhanced forestomach carcinogenesis. These results indicate that NaNO2 can modify phenolic antioxidant induced cell proliferation and/or carcinogenesis, particularly in the upper digestive tract. PMID- 8416748 TI - Mechanism of induction of c-fos by ultraviolet B (290-320 nm) in mouse JB6 epidermal cells. AB - The UVB (290-320 nm) portion of the solar spectrum possesses the highest activity for the induction of skin cancer and has the capacity to stimulate epidermal proliferation. We report that UVB is a transcriptional inducer of the c-fos protooncogene in mouse JB6 epidermal cells. Induction is biphasic with an immediate early peak at 30-60 min and a second broader peak 8 h following irradiation. The immediate early phase is suppressed by inhibitors of nuclear adenosine diphosphoribose transferase. For UVB induction, the formation of full length messages is less efficient than of early, short messages, while both types of messages are produced at similar rates following serum stimulation. Experiments with stable transfectants with reporter constructs linked to 5' upstream sequences of c-fos indicate that UVB and serum stimulation both require the sequences from -345 to -285 which contain the joint DSE-AP-1 enhancer motifs for efficient induction. Mobility shift data reveal that the complement of c-Fos and c-Jun proteins which bind to the fos-AP-1 octanucleotide decrease immediately following irradiation. Increased binding of Fos and Jun is observed 8-24 h later. UVB did not cause an observable change in the nuclear proteins which bind to the dyad symmetry element oligonucleotide in vitro. Fos protein was detected among the binding proteins. We propose that the two phases of UVB-induced c-fos expression occur by quite different mechanisms. The immediate early phase is inhibited by adenosine diphosphoribose transferase inhibitors because poly-ADP ribosylation of chromosomal proteins is required for the resealing of UVB-induced DNA strand breaks which otherwise retard message elongation. The production of an autocrine factor may be responsible for the late phase of c-fos induction. PMID- 8416749 TI - Caloric restriction and intervention in pancreatic carcinogenesis in the rat. AB - In two experiments, the effects of caloric restriction during the postinitiation phase of pancreatic carcinogenesis were evaluated. Male Lewis rats were given injections of azaserine at 14 days of age and weaned to the postinitiation test protocols at 21 days of age. In the first experiment, the caloric content of the diets was restricted by 10, 15, 20, and 30% of the intakes of the ad libitum-fed rats. A sixth group was fed diet ad libitum for only 5-6 h/day; i.e., they were "meal-fed". The development of putative preneoplastic lesions (henceforth termed foci) was evaluated by quantitative stereological (morphometric) analysis of the pancreas. Caloric restriction during the 4-month postinitiation phase resulted in a significant reduction in focal development beginning at 10% caloric restriction and increasing with more severe restriction. The caloric intake of the meal-fed group closely matched the caloric intake of the 10 or 15% caloric restriction groups and the focal response of the meal-fed rats was similar to the groups restricted in calories by 15 to 20%. In the second experiment, rats were initiated with azaserine and weaned to one of four groups: ad libitum; meal-fed; meal-fed for 2 months and ad libitum thereafter; or ad libitum for 2 months and meal-fed thereafter. Foci were evaluated at 2 and 4 months; neoplasm incidence and multiplicity were determined at 14 months postinitiation. Compared to the ad libitum group, the meal-fed group had significantly fewer foci at all times of evaluation and significantly fewer neoplasms. When rats were meal fed for 2 months and then switched to ad libitum feeding for the remainder of the experiment, the focal outcome at 4 months was similar to the group meal fed for all 4 months; and at 14 months the neoplastic outcome was intermediate between the ad libitum and the meal-fed group. Intervention in the ad libitum feeding regimen at 2 months by meal feeding for the remainder of the experiment resulted in a significant decrease in the focal and neoplastic development, as compared to the group fed ad libitum continuously. These two intervention groups were intermediate in response between the meal-fed and ad libitum-fed groups. These results indicate that the postinitiation phase of pancreatic carcinogenesis can be modulated by relatively simple dietary interventions such as moderate caloric restriction. PMID- 8416750 TI - Differential expression of MAGE-1, -2, and -3 messenger RNA in transformed and normal human cell lines. AB - The MAGE-1 gene codes for a tumor-specific antigen, MZ2-E, that elicited a cytotoxic T-lymphocyte response in the melanoma patient from whom it was derived. We have developed a simplified method, using polymerase chain reaction amplification of exon 3 followed by restriction enzyme pattern analysis, to distinguish expression of the MAGE-1 gene from MAGE-2 and MAGE-3, other members of this gene family. MAGE-1 mRNA was expressed in 53% of 17 melanoma lines, two of seven Epstein-Barr virus-transformed B-cell lines, and 2 of 5 breast cell lines including a line established form normal breast epithelium. MAGE-1 is not likely to be the common melanoma antigen recognized by the other HLA-A1- or HLA A2-restricted cytotoxic T-lymphocytes examined in this study, but the fact that it is expressed in about 50% of melanoma cell lines makes it a reasonable target for the immunotherapy of patients bearing HLA-A1. PMID- 8416751 TI - Wavelength dependence of skin cancer induction by ultraviolet irradiation of albino hairless mice. AB - Information on the variation in carcinogenicity with wavelength is crucial in risk assessments for skin cancers induced by UV radiation. Until recently the wavelength (lambda) dependencies of other detrimental UV effects, such as sunburn, have been used as substitutes. Direct information on the lambda dependency can only be obtained from animal experiments. To this end we accumulated a large data set on skin tumors induced by chronic UV exposure of albino SKH:HR1 mice (14 different broadband UV sources and about 1100 mice); the data come from the Photobiology Unit of the former Skin and Cancer Hospital in Philadelphia and from the Department of Dermatology of the University of Utrecht. The lambda dependency was extracted from this data set (a statistically satisfactory description with chi 2 = 13.4, df = 7) and represented by the Skin Cancer Utrecht-Philadelphia action spectrum, i.e., a set of factors to weight the exposures at different wavelengths according to their respective effectiveness (inversely proportional to the daily exposure required for a median tumor induction time of 300 days). The fits obtained with other already available action spectra proved to be poor (chi 2 > 60, df = 11). The maximum effectiveness was found at 293 nm, and above 340 nm the effectiveness showed a shoulder at about 10(-4) of the maximum. A sensitivity analysis of the final solution for the lambda dependency showed a large margin of uncertainty above 340 nm and an information gap below 280 nm. The large variation in tumor responses in the present data set can be transformed to a coherent, common dose-response relationship by proper spectral weighting with this single action spectrum. PMID- 8416752 TI - A prospective cohort study on dietary fat and the risk of postmenopausal breast cancer. AB - In 1986 a prospective cohort study on diet and cancer was started in the Netherlands among 62,573 women ages 55-69 years. Baseline information on diet and other risk factors was collected with a questionnaire. Cancer incidence was measured by record linkage with cancer registries and a pathology register. A case-cohort approach was used, in which the accumulated person time in the cohort was estimated by follow-up of a randomly selected subcohort (n = 1812). After 3.3 years of follow-up, 471 incident breast cancer cases were available for analysis. Questionnaire data for these cases and the 1716 female subcohort members without a history of cancer other than skin cancer were analyzed. In a multivariate analysis, controlling for traditional risk factors, the relative rates for breast cancer in increasing quintiles of energy-adjusted total fat intake were 1.00, 1.00, 1.34, 1.22, 1.08 (P-trend, 0.32). For saturated fat there was some evidence for a weak positive association when quintiles were used (relative rates in quintiles 1-5, 1.00, 1.22, 1.22, 1.38, 1.39; P-trend, 0.049). The 95% confidence interval (CI) for the top quintile was 0.94-2.06, however; and when saturated fat was used as a continuous variable, the effect was no longer significant (P = 0.20). Relative rate estimates for the highest versus lowest quintiles of monounsaturated fat, polyunsaturated fat, and cholesterol intake were 0.75 (95% CI, 0.50-1.12), 0.95 (95% CI, 0.64-1.40) and 1.09 (95% CI, 0.74-1.61), respectively, with no evidence for significant trends. This prospective study does not support a major role of dietary fat in the etiology of postmenopausal breast cancer. PMID- 8416753 TI - What is syndrome X? PMID- 8416754 TI - Evaluation of myocardial perfusion defects by means of "bull's eye" images. AB - The purpose of this study was to evaluate the usefulness of so-called "bull's eye" imaging as a simplified display of tomographic slices in the detection of coronary artery disease (CAD). A total of 64 patients were studied at stress and at rest, by single photon emission computed tomography (SPECT), either with thallium-201 (201Tl) or with technetium-99m methoxy-isobutyl-isonitril (Tc-MIBI). The myocardial perfusion defects detected by bull's eye image alone and in combination with visual analysis of tomographic images were evaluated in all cases, taking coronary arteriographic results as a gold standard. The overall sensitivity and specificity for detection of CAD were as follows: bull's eye imaging, 100% and 70.8%; tomographic imaging (SPECT), 90% and 91.6%. The results of bull's eye imaging and SPECT interpreted together were 96.6% and 83.3%. The regional sensitivity and specificity of bull's eye for individual coronary arteries were: right coronary artery (RCA), 100% and 73.7%; left anterior descending (LAD), 100% and 87.2%; left circumflex (LCx), 100% and 97.3%. For SPECT they were: RCA, 93.7% and 89.5%; LAD, 86.6% and 92.3%; LCx, 73.3% and 97.4%. For bull's eye with SPECT they were: RCA, 94.4% and 86.1%; LAD, 87.5% and 92.1%; LCx, 82.3% and 97.2%. We conclude that the bull's eye image display allows an easier and more objective assessment of myocardial perfusion defects and shows higher sensitivity. However, it has a relatively low specificity which can cause an overestimation of perfusion defects. Thus, visual analysis of bull's eye imaging is a useful diagnostic tool but must be evaluated in conjunction with tomographic imaging. PMID- 8416755 TI - Usefulness of CKMB and troponin T determinations in patients with acute myocardial infarction complicated by ventricular fibrillation. AB - The usefulness of creatine kinase (CK) time activity curves for diagnosis of acute myocardial infarction in patients who have been defibrillated for ventricular fibrillation is limited due to the release of the enzyme as a result of the countershock. The present study of four patients with acute infarction complicated by primary ventricular fibrillation indicates that analysis of CKMB and even more so of troponin T, a specific cardiac antigen, permits reliable diagnosis of acute myocardial infarction in this particular setting. Furthermore, the data indicate that noninvasive assessment of coronary artery patency may be also possible in these patients by means of these two serum markers. PMID- 8416756 TI - Duplex sonographic detection of internal jugular venous thrombosis after removal of central venous catheters. AB - Venous thrombosis is a very common complication of central venous catheters inserted via the jugular or subclavian vein. The aim of this prospective study is to test the suitability of duplex sonography in detecting thrombi in the internal jugular vein after catheter removal and to analyze the number and extent of such thrombus formations. The study group comprised 64 consecutive patients with an average age of 59 +/- 16 years admitted to an internal intensive care unit. In 54 cases the catheters were removed under sonographic control. In 10 cases duplex sonography was carried out within 24 h after removal of the catheters. The data of 63 patients were evaluated. In 40 patients (63.5%), thrombi of varying extent were present in the jugular vein. No significant correlations were found between thrombus formation and the basic disease, duration of cannulation, the type of catheters used, and the mode of heparinization (therapeutic vs. low-dose). Local inflammation signs and local hematoma showed a significantly higher incidence (p < 0.01) in patients with thrombus formation. Duplex sonography lends itself as a valuable diagnostic tool in the diagnosis of thrombus formation in the internal jugular vein after removal of central venous catheters. Sonographic examinations should be carried out in all long-stay patients at an intensive care unit, in whom central venous catheters are inserted repeatedly via the internal jugular vein. PMID- 8416757 TI - Mitral regurgitation following first-time acute myocardial infarction--early and late findings by Doppler echocardiography. AB - A total of 61 patients with first-time mild to moderate acute myocardial infarction and no reinfarction within the following 2 months were studied prospectively by Doppler echocardiography before hospital discharge and after 2 months to evaluate the prevalence of mitral regurgitation. Twenty-one age-matched healthy subjects served as controls. At baseline, the prevalence of Doppler recorded mitral regurgitation was 74% and 29% in patients and controls, respectively. In the patients, the regurgitant flow measured by color flow Doppler was 1.04 cm2 (range 0.2-8 cm2) and occupied 7.5% (range 2-45%) of the left atrial area. Corresponding figures for controls were 0.35 cm2 (0.1-0.6) and 2.4% (0.7-4.5), respectively. On continuous wave Doppler, most patients (33/45) had Doppler signals similar to those of healthy controls. The prevalence of mitral regurgitation was about the same in anterior and inferior infarction (75 and 72% respectively). In the patients, the prevalence was similar after 2 months (79%) with minor changes in the Doppler characteristics of the regurgitation (regurgitant flow 1.12 cm2 and occupying 8.1% of left atrial area). The study demonstrates that in a group of patients with first-time mild to moderate myocardial infarction the prevalence of Doppler-recorded mitral regurgitation is high and mild in severity in the majority of the cases. The changes remain almost similar even after 2 months. PMID- 8416758 TI - Abnormal QT intervals associated with negative T waves induced by antiarrhythmic drugs are rapidly reduced using magnesium sulfate as an antidote. AB - This study was undertaken to determine whether prolonged QTc interval as a consequence of abnormal repolarization induced by coronary disease or antiarrhythmic drugs could be shortened by intravenous administration of magnesium sulfate. A total of 21 patients with basal prolonged QTc intervals (QTc > 500 ms) were divided in two groups: 7 with ischemic coronary disease and negative T waves (Group A), and 14 treated with antiarrhythmic drugs (Group B). Nine of the latter had negative T waves (Subgroup B-1) and five had positive T waves (Subgroup B-2) recorded in precordial leads. Nine patients were taking amiodarone and six quinidine. Magnesium sulfate was given intravenously in a bolus of 3.75 g (25% solution) over 3 min. Patients had normal electrolyte serum levels. The prolonged QTc and JTc intervals were shortened after magnesium sulfate in patients of Subgroup B-1 from the basal values [QTc 20.7% and JTc 25.4%, (p = < 0.0001 and 0.02, respectively)]. None of the patients in Group A or Subgroup B-2 experienced altered QTc or JTc intervals. While some antiarrhythmic drugs are capable of altering the refractoriness of ventricular cells, probably by causing changes in the intracellular metabolic pathways, in patients with coronary disease gaps in the membrane induced by ischemic injury let calcium enter the cells parallel with dispersion of ventricular repolarization. When secondary negative T waves are present, magnesium sulfate as an antidote probably acts as a blocking agent at the sarcoplasmic reticulum, thus reducing both QTc and JTc intervals. PMID- 8416759 TI - Asymmetrical septal hypertrophy in patients with hypertension: a type of hypertensive left ventricular hypertrophy or hypertrophic cardiomyopathy combined with hypertension? AB - To determine whether asymmetrical septal hypertrophy (ASH) in patients with essential hypertension (HT) is a type of hypertensive left ventricular (LV) hypertrophy or hypertrophic cardiomyopathy (HCM) combined with HT, we investigated a group of 7 hypertensive patients with ASH compared with 12 HCM patients and 10 healthy controls using radionuclide angiography and right ventricular endomyocardial biopsy. The LV time-volume curve and its first and second derivative curves were constructed from cardiac output and time-activity curves constructed by combined forward and reverse-gating from the R wave. The LV wall thickness and ejection fraction were significantly greater in both the HT and HCM groups than in the control group, whereas there were no differences in these indices between the HT and HCM groups. Rapid filling volume index and rapid filling fraction showed significantly lower values in the HCM group than in the control group (p < 0.005). In contrast to the HCM group, these indices in the HT group did not differ from those in the control group. The time to peak filling rate was prolonged in the control, hypertension, and HCM groups in increasing order. Histopathological study revealed a higher incidence of myocardial cell disarray in the HCM than in the HT group. The above results suggest that ASH in hypertensive patients is a type of hypertensive LV hypertrophy. PMID- 8416760 TI - Combination of ramipril and hydrochlorothiazide in the treatment of mild to moderate hypertension--Part 2: An open long-term study of efficacy and safety. AB - In an open, multicenter extension of a short-term study, 159 patients with mild to moderate hypertension were treated with either ramipril monotherapy or a combination of ramipril and hydrochlorothiazide for up to 1 year. Patients started with either 5 mg ramipril once daily (responders in the short-term study) or a combination of ramipril 5 mg plus hydrochlorothiazide 25 mg once daily. The dose could be adjusted and nonresponders to ramipril monotherapy could have hydrochlorothiazide added. In the 38 patients treated with ramipril monotherapy, the largest drop in mean blood pressure (BP) had already occurred in the previous short-term study; from Week 2 in the long-term study, the BP remained stable with means below 150/90 mmHg. In the 83 patients treated with the combination for 50 weeks or more, mean BP continued to decrease until around Week 10 in the long term study while therapy was being adjusted. Thereafter, it also remained stable with means below 150/85 mmHg. Both treatment groups showed good mean reductions at end point, as did the group of 38 patients treated with the combination for less than 50 weeks. High response rates (84-95%) were seen in all groups at end point. The combination was well tolerated and the efficacy of ramipril in combination with hydrochlorothiazide was maintained over the 1-year period of investigation. PMID- 8416761 TI - Inotropic therapy of the failing myocardium. AB - The clinical syndrome of congestive heart failure remains a therapeutic dilemma and challenge for the physician in 1992. This is a disease process that appears to be increasing in frequency and continues to carry an unacceptably high mortality rate. For years it has been well recognized that the combination of digoxin, Lasix and vasodilator therapy improved symptoms in these patients and decreased hospitalization, but did not increase survival. It was not until 1986 that the combination of digoxin, Lasix, Isordil, and hydralazine was shown to increase survival. Further significant improvement in quality of life and survival has recently been established in three large clinical trials, and it is now safe to say that the standard of care for symptomatic congestive heart failure in 1992 is digoxin, furosemide, and an ACE inhibitor, with the survival trials favoring the ACE inhibitor enalapril. The IV inotropic drug dobutamine remains the mainstay of pharmacological therapy for the treatment of severely refractory heart failure. Unfortunately, the phosphodiesterase inhibitors- amrinone, milrinone, and enoximone--have demonstrated unacceptable clinical side effects and have been withdrawn from further clinical study. In spite of these promising developments, the mortality and morbidity of congestive heart failure remains unacceptably high, and continued investigation in the new fields of pharmacology and the pathophysiology of congestive heart failure still must be aggressively pursued. PMID- 8416762 TI - Long-term double-blind evaluation of amlodipine and nadolol in patients with stable exertional angina pectoris. The Investigators of Study 152. AB - The efficacy and tolerability of amlodipine 2.5-10 mg once daily was compared with nadolol 40-160 mg once daily in a long-term double-blind parallel-group study in patients with stable exertional angina pectoris. A total of 80 patients were randomized to receive amlodipine or nadolol for 26 weeks after a 2-week single-blind placebo run-in period. The effects of amlodipine and nadolol on total exercise time were minimal and not significantly different. However, amlodipine produced a slightly but not significantly greater increase in time to onset of angina than nadolol (+21% amlodipine; +8% nadolol). No significant differences were noted between amlodipine and nadolol on ST-segment depression, angina attack rate, or nitroglycerin consumption. A slightly greater improvement was attained after amlodipine on patient and investigator assessments of treatments. A statistically significant difference (p < 0.0001) was found between treatments on the effects on myocardial oxygen requirements (as assessed by the rate pressure product). Nadolol produced a reduction of 29% compared with a slight reduction of 4% with amlodipine. Fewer side effects were reported with amlodipine (43%) than with nadolol (83%) (p < 0.0001), resulting in discontinuation of therapy in three amlodipine and four nadolol patients. Long term treatment with amlodipine and nadolol produced comparable effects in patients with angina pectoris, with fewer side effects being reported after treatment with amlodipine. PMID- 8416763 TI - Acute myocardial infarction with papillary muscle rupture. AB - The subject of this report is a 57-year-old obese, hypertensive woman who had been well until the onset of severe chest pain and hypotension. She had to be defibrillated four times on her way to the hospital. The diagnosis of acute inferior-posterior infarction was made by electrocardiogram (ECG) and there was a markedly elevated serum creatine kinase (CK) (including the MB fraction). The patient had a very low cardiac output and ejection fraction. A lung scan revealed possible pulmonary embolism for which she was anticoagulated. She remained hypotensive and hypoxemic and, on Day 17 of her hospital stay, she had a bout of severe dyspnea. A new systolic murmur was heard and the clinical diagnosis of ruptured papillary muscle was made and confirmed by echocardiography, and later at autopsy. All three coronary arteries were severely atherosclerotic and, in addition, the right coronary artery was completely closed by a thrombus. This case clearly illustrates the major pathological changes in the heart that correlate with the clinical findings in patients with a myocardial infarct that is complicated by left ventricular papillary muscle rupture. The pathophysiological effects of this condition, as illustrated in this case report, include the following:1. The posterior papillary muscle wa s almost completely separated from its base, with only a thin strip of muscle intact. The mistral valve thus was insufficient (a "flail valve''); this markedly reduced the ejection fraction of the left ventricle, increased its end-diastolic volume and pressure, produced a damming of blood in the pulmonary circulation, and this resulted in the pulmonary edema seen on the chest x-ray.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8416764 TI - Effects of physical exercise training in syndrome X. AB - Syndrome X categorizes myocardial disease characterized by impaired myocardial flow without macrovessel injury. The pathogenesis of this condition remains obscure. Similarly, therapy is not well defined. We report the effects of physical exercise training in a patient with Syndrome X. PMID- 8416765 TI - Inadvertent thrombolytic therapy for cardiovascular diseases masquerading as acute coronary thrombosis. AB - We report three cases of inadvertent thrombolytic administration to patients with cardiovascular diagnoses masquerading as acute coronary thrombosis presenting to tertiary care private hospital. Despite a final diagnosis of myocarditis, aortic dissection, and pericarditis, the initial presentation and electrocardiogram were believed to indicate an acute myocardial infarction due to coronary thrombosis. Intravenous thrombolytic agents were administered early in their presentation. Cardiac catheterization in two of the patients revealed normal coronary arteriography and in the third patient confirmed an aortic dissection. The patient with an aortic dissection died while the other two recovered without adverse consequences of the thrombolytic agents. Prior reports of five patients, treated with intravenous thrombolytic agents for suspected coronary thrombosis, who proved to have a final diagnosis of pericarditis or aortic dissection are reviewed. Death or tamponade occurred in four of five. The consequences of inadvertently administering intravenous thrombolytic agents to patients with nonthrombolytic cardiac disorders can be serious. If the diagnosis of acute myocardial infarction due to coronary thrombosis is uncertain, serial electrocardiograms, bedside echocardiography, or urgent cardiac catheterization may be appropriate before administering these agents. PMID- 8416766 TI - Infective endocarditis due to Fusobacterium nucleatum: case report and review of the literature. AB - Endocarditis caused by Fusobacterium nucleatum is exceedingly rare. We report a case of F. nucleatum right-sided endocarditis in a patient with no known intravenous drug abuse. A detailed computer and manual search of the literature using Med-Line and Index Medicus, respectively, revealed a total of five cases of endocarditis due to this species. The clinical characteristics and the patient's hospital course are summarized and reviewed. PMID- 8416767 TI - Albion Walter Hewlett. PMID- 8416768 TI - Detection and localization of early lung cancer by imaging techniques. AB - Two new technologic developments may have a significant impact on the detection and localization of early lung cancer. These two developments work together in a complementary way. The first is a solid-state microscope that can be applied in the prescreening of sputum cytology specimens. The finding that malignancy associated changes (MACs) are present in ostensibly normal bronchial epithelial cells may be used to improve the sensitivity of sputum cytology to detect cancer. Once abnormal or MAC cells are found, a second device, a fluorescence bronchoscope, can be employed to localize the source of the abnormal cells. Fluorescence bronchoscopy is also a potentially useful tool for procuring premalignant tissue for molecular biology studies and for monitoring the progress of patients in chemoprevention studies. PMID- 8416769 TI - Chemoprevention strategies in lung carcinogenesis. AB - Chemoprevention entails using specific agents to suppress carcinogenesis and thereby prevent the development of primary or second primary cancers. Because the concept of chemoprevention in patients with or at risk of lung cancer is new, ongoing clinical trials are based on data from epidemiologic and preclinical research, as well as on results of chemoprevention studies in head and neck cancer. The latter studies have provided a model for such studies in lung cancer, considering the two diseases have a similar etiology and biology of field carcinogenesis. Beta-carotene, natural vitamin A, and the retinoids may be effective chemopreventive agents. However, chronic administration of such agents may be required to prevent the development of cancer. Results of chemoprevention trials in head and neck cancer have demonstrated effective inhibition of the development of second primary tumors with the synthetic retinoid 13-cis-retinoic acid; investigators are hopeful this will be repeated in patients with lung cancer. Results of ongoing phase III trials and continued advances in the epidemiologic and biologic study of lung carcinogenesis should contribute to future research in this area. PMID- 8416770 TI - The epidemiology of lung cancer. AB - Lung cancer rates and mortality have risen in epidemic proportions in the United States and other industrialized nations during the 20th century. Case-control and cohort studies performed in the 1950s and 1960s firmly established cigarette smoking as the single greatest risk factor for lung cancer. In the United States, overall lung cancer mortality rates in men and women rose progressively from the 1950s. Fortunately, lung cancer incidence and mortality are now declining in middle-aged men. Smoking has significantly increased lung cancer rates among women and is on the rise in developing countries. Environmental agents found in the home and workplace, including radon and asbestos, have also been shown to increase lung cancer risk in both smokers and nonsmokers. Government regulations have helped curtail quantities of these and other atmospheric carcinogens. Efforts to reduce lung cancer risk must be continued and their scope expanded in order to have a global impact on the incidence and mortality of this fatal malignancy. PMID- 8416771 TI - Chemoprophylaxis strategies in high-risk groups with an emphasis on lung cancer. AB - The incidence of lung cancer in the United States has stabilized in recent years, but it remains a major cause of death in the United States. Whereas the single most effective primary prevention of this disease would be to eliminate tobacco use from society, this is currently an unrealistic goal. Secondary prevention, however--that is, chemoprophylactic treatment of smokers, exsmokers, and others at risk--represents a viable option. Agents proven effective in both laboratory models and humans include vitamin A and its synthetic derivatives, the retinoids and the carotenoids. It is fairly easy to identify patients at risk of lung cancer compared with other cancers. Yet aside from patients who are under a physician's care and aware of their risk, it can be difficult to target individuals for chemoprophylactic treatment, especially those who are healthy but at high risk and not seeing a physician or other health care provider. Screening for the presence of predictive cellular and molecular changes may facilitate more accurate selection of individuals for chemoprophylactic treatment. PMID- 8416772 TI - Surveillance in hereditary nonpolyposis colorectal cancer: an international cooperative study of 165 families. The International Collaborative Group on HNPCC. AB - During its second meeting at Amsterdam in 1990, the International Collaborative Group on Hereditary Non-Polyposis Colorectal Cancer (ICG-HNPCC) decided to carry out a pilot study on colorectal cancer surveillance in HNPCC. The objectives of the study were to ascertain in each of the participating centers the number of HNPCC families, the recommended screening procedures, the age at diagnosis of colorectal cancer (CRC), and the occurrence of interval cancers. Nine centers in seven countries including Denmark, Finland, Italy, Japan, The Netherlands, Switzerland, and the United States participated. Data were derived from a total of 165 families. With respect to screening, half of the centers advise colonoscopy as the only procedure. The interval between the consecutive examinations varies from one to three years. In the majority of the centers, screening begins at 20 to 25 years. Lifelong screening is recommended by three centers, while the rest advise discontinuation at age 60 to 75 years. The family material included 840 patients with colorectal cancer. The mean age at diagnosis was 45 years, and about 15 percent were diagnosed at age 60 or later. A total of 682 high-risk relatives are being followed. After the follow-up of 1 to 10 years in these families, only six cases of interval cancers were encountered. PMID- 8416773 TI - Arrangement of the anal striated musculature. AB - A study of the anatomy of the anal striated musculature is performed on 34 adult specimens to reconsider some debated morphologic aspects. The original "eight shaped" arrangement of the fibers of the pubococcygeus is pointed out, suggesting a definite role of the muscle as a superior part of the continence muscular complex; the intermediate part seems to be formed by the puborectalis loop and the deep external anal sphincter, not always separable. Finally, the inferior portion is represented by the superficial sphincter, which shows anatomic behavior similar to that of the pubococcygeus. PMID- 8416774 TI - Mechanism of sphincter impairment after stapled low anterior resection is questioned. PMID- 8416775 TI - Creation of a pedicle valve unit (PVU) for establishment of enteric continence. Experimental observations. AB - The aim of this study was to develop a natural tissue valve that could be anastomosed into any area of the gastrointestinal (GI) tract to act as a fecal "brake" and so establish enteric continence at that site. A 4-cm-long valve created from an intussuscepted small bowel pedicle was anastomosed into the cecum and brought out through the abdominal wall as a stoma in 11 rabbits. The animals were re-explored five weeks later for assessment of valve viability and continence and microscopic appearance. In all cases, the valve was fully continent in vivo. All valves were viable, and there was no anastomotic leakage. Pressure testing of the valve at reoperation revealed that 7 of 10 valves tested withstood pressures of 30 mmHg before and after catheterization and 6 of 10 were fully continent to cecal "blanching" pressure (50 mmHg). Valve failure was due to deintussusception in three cases. In four cases, valves were continent over 50 mmHg and showed no tendency to incontinence to bursting pressure of the cecum. We conclude that a continent pedicle valve unit (PVU) for placement in a variety of locations in the GI tract is feasible. The PVU has implications in the management of short-gut syndrome, incontinent ileostomy, continent cecostomy, and as a continent valve placed in the perineum for restoration of perineal defecation following proctectomy. PMID- 8416776 TI - Laparoscopic colectomy: a critical appraisal. AB - A multicenter retrospective study was undertaken to assess the efficacy and safety of laparoscopy in colon and rectal surgery. To minimize potential bias in interpretation of the results, all data were registered with an independent observer, who did not participate in any of the surgical procedures. Sixty-six patients underwent a laparoscopic procedure. Operations performed included sigmoid colectomy (19), right hemicolectomy (15), low anterior resection (6), colectomy with ileal pouch-anal anastomosis (IPAA) (5), and abdominoperineal resection (APR) (3). The conversion rate from laparoscopic colectomy to celiotomy was 41 percent. Major morbidity and mortality were 24 percent and 0 percent, respectively. Length of stay, hospital costs, and lymph node harvest were compared between the sigmoid resection and right hemicolectomy subgroups. Data from traditional sigmoid colectomies and right hemicolectomies were obtained from the same institutions for comparison. Mean postoperative stay for laparoscopically completed sigmoid and right colectomies was significantly less than that for either the converted or the traditional groups (P < 0.02). Total hospital cost for traditional right hemicolectomy was significantly less than that for the converted group (P < 0.05) but not the laparoscopic group. Laparoscopic sigmoid resection showed no significant total hospital cost difference among traditional, converted, and laparoscopic groups. Lymph node harvest in resections for carcinoma was comparable in all groups. These preliminary data suggest that laparoscopic colon and rectal surgery can be accomplished with acceptable morbidity and mortality when performed by trained surgeons. Length of stay is shorter, but there is no proven total hospital cost benefit. Appropriate registries will be necessary to adequately assess long-term outcome. PMID- 8416777 TI - Fecoflowmetry: a new parameter assessing rectal function in normal and constipated subjects. AB - Fecoflowmetry is a new technique by which the fecal flow rate is studied through recorded curves representing the changes that occur in the flow against time. Fecal flow rate is the product of rectal detrusor action against outlet resistance. The technique was performed on 36 normal volunteers and 88 chronically constipated patients. Simultaneous recording of the fecal flow rate and intra-abdominal and rectal neck pressures were performed. A water or paste enema was given to the individual. Upon feeling the desire to defecate, he or she was placed on a fecoflowmeter commode and was asked to defecate. Evaluation of the obtained defecation flow curve comprises the reporting on the defecated volume, flow time, mean and maximum flow rates, time to maximum flow, and shape of the curve. In the 88 constipated patients, two fecoflowmetric patterns were recognized: nonobstructive (inertia) and obstructive. They differ from each other in parameters and curve configuration. The defecated volume as well as mean and maximum flow rates were lower in outlet obstruction than in the inertia type, whereas flow time and time to maximum flow were longer. The ascending limb in the obstructive-type curve rose less steeply than in inertia; the curve had a long plateau, and the descending limb sloped more gradually. To conclude, fecoflowmetric studies could differentiate between defecation of normal and constipated subjects, and in the latter between the obstructive and inertia types of constipation. The technique was developed to simulate natural defecation. It provides quantitative and qualitative data concerning the defecation act. The technique is simple, easy, noninvasive, and nonradiologic. It can be used as a screening tool in defecation disorders. PMID- 8416778 TI - Clinical studies in surgical journals--have we improved? AB - A critical appraisal of all clinical studies published in 1980 and 1990 in three surgical journals--Diseases of the Colon and Rectum (DCR), Surgery (SURG), and the British Journal of Surgery (BJS)--was made to ascertain the frequency with which various research designs appeared, the standard of individual clinical studies, and a comparison of changes in the past decade. Clinical studies were classified into case studies or comparative studies. Comparative studies included randomized controlled trials (RCT), nonrandomized controlled trials, retrospective cohorts, and case-control studies. A 10-point index score (range, 0 10) was used to assess the comparative studies. A sample of articles was analyzed for interobserver and intraobserver variation, with strong agreement between reviewers for classification of trials (unweighted kappa, 0.87) and index scores (0.67). Of 1,481 articles reviewed, 1,060 were classified as clinical studies. Sixteen percent of all clinical studies were comparative studies in 1980, compared with 17 percent in 1990. Of these, 7 percent were RCT in both years. In 1980, 6 percent of clinical studies in DCR were comparative studies, 19 percent in BJS, and 18 percent in SURG. In 1990, 11 percent, 18 percent, and 18 percent, respectively, were comparative studies. In 1980, the proportion of RCT in DCR was 0 percent, in BJS 12 percent, and in SURG 4 percent, compared with 3 percent, 8 percent, and 8 percent, respectively, in 1990. Overall, 52 of 76 (68 percent) RCT were published in BJS. The standard of comparative studies increased overall from 5.49 to 6.04 (P = NS), and that of RCT increased from 7.06 to 7.70 (P = NS). The standard of comparative studies in DCR in 1980 was lower than those in BJS (P < 0.001) and SURG (P < 0.001). The standard of comparative studies in DCR improved from 1.67 in 1980 to 5.47 in 1990 (P < 0.001). There was no significant difference in the standard of comparative studies among the three journals in 1990. In conclusion, there has been no overall increase in the proportion of stronger clinical trial designs in the journals reviewed. A small increase seen in the overall standard of comparative studies was not statistically significant. PMID- 8416779 TI - Clinicopathologic correlations of ABO and Rhesus blood type in colorectal cancer. AB - There is no specific association established between colorectal cancer and blood group type. In this study, the distribution of ABO and Rh blood groups was studied in 838 patients with colorectal cancer. There was no difference in distribution of ABO blood groups between patients who were Rh+ and Rh-. There was no difference in ABO blood group or Rh factor and tumor location. The highest A/O ratio was found in rectal cancer. Although there was no difference in stage distribution for each ABO blood group, there was a significant difference between the Rh+ and Rh- groups (P < 0.037). It is not clear, however, whether the prognosis is different between the two groups since there were more early tumors as well as incurable tumors in the Rh- group. All patients with synchronous cancer were Rh+. Further studies on blood group antigens are needed to elucidate the relationship between these antigens and colorectal cancer. PMID- 8416780 TI - Cerebral potentials evoked by electrical stimulation of the anal canal. AB - We describe the procedure with which cortical potential responses are evoked by a stimulation of the anal canal to assess the integrity of its sensory pathways. These potentials were recorded in 66 patients. In 44 patients, a cortical evoked response was obtained with a succession of positive and negative peaks, W shaped (35 cases) or V shaped (nine cases). In seven cases, cortical responses were interpreted differently by two independent observers. In these seven patients, such differences could be explained by an insufficient amplification of the recorded electrical waves recorded on paper (< 10 mm). Fifteen patients gave no cortical response. Eight had a neurologic disease that could explain the lack of response. In the seven others, the absence of response was considered as false negative, but six of these stimulations had been carried out during the first part of the study. There is some evidence that cortical evoked potentials may be obtained after an electrical stimulation of the anal canal, but a training period seems necessary to master the technique and obtain reproducible and recognizable responses. PMID- 8416781 TI - Diagnostic laparoscopy and laparoscopic cecostomy for colonic pseudo-obstruction. AB - Marked cecal dilatation due to colonic psuedo-obstruction (Ogilvie's syndrome) is most often treated by colonoscopic decompression. When this fails, cecostomy is usually indicated if the bowel is not infarcted. We describe a new technique of laparoscopy-guided percutaneous cecostomy using T-fasteners to retract and anchor the cecum to the anterior abdominal wall and using a Foley catheter as a cecostomy tube. We performed this procedure successfully in a patient with colonic pseudo-obstruction who had marked cecal dilatation that could not be decompressed by colonoscopy. Laparoscopic inspection showed that the cecum was viable, and a laparoscopic cecostomy was placed. This procedure can be performed easily and safely and with much less morbidity than laparotomy and open cecostomy. PMID- 8416782 TI - Strictureplasty for Crohn's disease with multiple long strictures. AB - Strictureplasty of both the Heineke-Mikulicz and Finney varieties has safely corrected certain small bowel strictures associated with Crohn's disease. A new technique incorporating both these varieties in a single strictureplasty has further expanded the technical repertoire of strictureplasty and is hereby described. PMID- 8416783 TI - Triangulating stapling technique: an alternative approach to colorectal anastomosis. AB - The triangulating stapling technique was employed to perform colorectal anastomosis in 259 patients. In 220 patients, the anastomosis was performed between the colon and nonperitonealized rectum. This anastomotic technique is safe and reliable and is an effective alternative to a circular stapling device, with minimal morbidity. The incidence of leak rate is comparable to anastomoses created by a circular stapling device. The main advantage seems to be the very low incidence of anastomotic stenosis. PMID- 8416784 TI - Etiology and management of fecal incontinence. AB - Fecal incontinence is a challenging condition of diverse etiology and devastating psychosocial impact. Multiple mechanisms may be involved in its pathophysiology, such as altered stool consistency and delivery of contents to the rectum, abnormal rectal capacity or compliance, decreased anorectal sensation, and pelvic floor or anal sphincter dysfunction. A detailed clinical history and physical examination are essential. Anorectal manometry, pudendal nerve latency studies, and electromyography are part of the standard primary evaluation. The evaluation of idiopathic fecal incontinence may require tests such as cinedefecography, spinal latencies, and anal mucosal electrosensitivity. These tests permit both objective assessment and focused therapy. Appropriate treatment options include biofeedback and sphincteroplasty. Biofeedback has resulted in 90 percent reduction in episodes of incontinence in over 60 percent of patients. Overlapping anterior sphincteroplasty has been associated with good to excellent results in 70 to 90 percent of patients. The common denominator between the medical and surgical treatment groups is the necessity of pretreatment physiologic assessment. It is the results of these tests that permit optimal therapeutic assignment. For example, pudendal nerve terminal motor latencies (PNTML) are the most important predictor factor of functional outcome. However, even the most experienced examiner's digit cannot assess PNTML. In the absence of pudendal neuropathy, sphincteroplasty is an excellent option. If neuropathy exists, however, then postanal or total pelvic floor repair remain viable surgical options for the treatment of idiopathic fecal incontinence. In the absence of an adequate sphincter muscle, encirclement procedures using synthetic materials or muscle transfer techniques might be considered. Implantation of a stimulating electrode into the gracilis neosphincter and artificial sphincter implantation are other valid alternatives. The final therapeutic option is fecal diversion. This article reviews the current status of the etiology and incidence of incontinence as well as the evaluation and treatment of this disabling condition. PMID- 8416785 TI - Clinical significance of diminutive polyps of the rectum and sigmoid colon. AB - A retrospective review of 637 consecutive colonoscopies with polypectomy in 526 patients was performed to determine the association of small polyps of the rectum and sigmoid colon with more proximal colonic neoplasms. All colonic polyps were proximal to the sigmoid colon in 117 procedures. Proximal neoplasms were found in 32 percent of patients with a single polyp in the rectum or sigmoid colon. The incidence increased to 83 percent for those with three or more polyps. The occurrence of proximal colonic neoplasms was not affected by the size or histologic type of the rectosigmoid polyps. These findings would suggest that total colonic evaluation be considered in all patients with a polyp in the rectum or sigmoid colon regardless of the size or histologic type of the polyp. PMID- 8416786 TI - What are the appropriate controls for a colonoscopic screening program? PMID- 8416787 TI - Theoretical considerations on the formation of secretory granules in the rat pancreas. AB - Rat pancreatic zymogen granule sizes were determined by analysis of electron micrographs of the pancreas from adult and newborn rats. Areas of granule profiles were measured and converted to equivalent volumes. Histograms of the equivalent volumes showed integral multimodal distributions which were evaluated for goodness of fit with two models, unit addition and random fusion. Previous analyses of zymogen granule size distributions have failed to recognize the multimodality we have observed. Distributions of equivalent volumes for the two models were developed using Monte Carlo simulation. In the case of the granules from the newborn rats, the distribution of granule sizes gave a better fit with the random fusion model, whereas the granules from the adult rats had distributions with a better fit to the unit granule addition model. The estimated unit granule sizes for the two different ages were the same. Both unit addition and random fusion models propose that following formation of secretory granules from Golgi-derived material, the granules fuse with one another to create a wide dispersion of granule sizes. The present results extend the evidence for fusional growth of secretory granules, originally developed for the mast cell, to the zymogen granules of pancreas. All normal cells previously studied have yielded secretory granule distributions most consistent with unit addition. The basis for the expression of random fusion in the newborn rather than the more usual unit addition is not known. PMID- 8416788 TI - Monitoring motion of confluent cells in tissue culture. AB - Cell movements in confluent fibroblastic cell layers have been followed with a new instrumental method. Using the electric cell-substrate impedance sensor (ECIS), cell motions on the scale of nanometers are manifested as fluctuations in the impedance of small gold film electrodes that serve as cell substrata. These fluctuations have been recorded and numerically processed for cells at 27 and 37 degrees C, for cells deprived of all external nutrients over extended periods of time, and for cells exposed to different levels of cytochalasin D. Results suggest that the movements in confluent layers detected via ECIS can be correlated with cell metabolic activity. PMID- 8416789 TI - Micromanipulation of chromosomes in PTK2 cells using laser microsurgery (optical scalpel) in combination with laser-induced optical force (optical tweezers). AB - An optical scalpel and optical tweezer have been combined to perform intracellular microsurgery and micromanipulation in vivo. When only laser microsurgery was performed on metaphase chromosomes, the dissected sister chromatid fragments drifted off to either the side of the spindle or completely off the spindle. At anaphase the fragments separated and the two arms generally moved to their respective daughter cells. When the chromosome arm was cut during anaphase A and B, the distal chromosome fragment separated from the rest of the chromosome and moved toward the pole, following the proximal chromosome fragment. Distal chromosome fragments laser-dissected during metaphase were held together throughout anaphase using the optical trap. Optical trapping of dissected chromosome fragments during anaphase A and B inhibited movement of the chromosome fragment to its pole. As a result, the trapped chromosome fragments were (1) incorporated into the opposite daughter cell, (2) lost in the cleavage furrow during cytokinesis, or (3) eventually incorporated into the correct daughter cell. These results indicate that optical traps are effective in holding laser dissected chromosome fragments throughout mitosis. This new tool should be useful for studies on chromosome movement and cell genetics. PMID- 8416790 TI - Type I collagen promotes modulation of cultured rabbit arterial smooth muscle cells from a contractile to a synthetic phenotype. AB - The phenotypic transition of smooth muscle cells (SMC) from a contractile to a synthetic state appears to be an early event in the pathogenesis of atherosclerosis. We examined the effects of extracellular matrix components on the phenotypic modulation of rabbit arterial SMC in primary culture by flow cytometry. The results demonstrate that freshly isolated SMC attached, spread, and started to proliferate on type I collagen as well as on fibronectin. Moreover, type I collagen was as efficient as fibronectin in promoting the transition of the cells into the synthetic phenotype without exogenous mitogens. However, unlike on fibronectin, the synthetic peptide GRGDSP (Gly-Arg-Gly-Asp-Ser Pro) and the peptide KDGEA (Lys-Asp-Gly-Glu-Ala), which contains the recognition sequence for alpha 2 beta 1 integrin in type I collagen, interfered little with the attachment, spreading, and phenotypic modulation of the cells on type I collagen. On the other hand, the phenotypic modulation of the cells was counteracted by the anti-beta 1 integrin antibody. These findings indicate that type I collagen promotes the phenotypic transition of the rabbit arterial SMC by interacting with a cell surface receptor (beta 1 integrin family) for a cell binding sequence without RGD and DGEA. In contrast, elastin, a major constituent of the media, suppressed the cell attachment and spreading and maintained the cells in the contractile phenotype as laminin. These results suggest diverse roles of type I collagen and elastin as well as of fibronectin and laminin in the control of the differentiated properties of arterial SMC. PMID- 8416791 TI - AP-1 activity during normal human keratinocyte differentiation: evidence for a cytosolic modulator of AP-1/DNA binding. AB - Increased levels of c-fos and c-jun expression have been observed in differentiating epithelial cells. However, no data are available on activator protein 1 (AP-1) activity during keratinocyte differentiation. In this work we investigated c-fos and c-jun gene expression and AP-1-(12-O-tetradecanoylphorbol 13-acetate)-responsive enhancer element (TRE) binding activity during keratinocyte differentiation utilizing both authentic and in culture reconstituted human epidermis. We demonstrate that: (i) in reconstituted epidermis, non-differentiated and differentiated keratinocytes express equivalent levels of c-Jun, while in reconstituted epidermis permanently grafted onto athymic mice, as well as in authentic epidermis, c-Jun is predominantly expressed in the granular layer of the tissue. Equivalent levels of c-fos expression have been found in all the layers of both reconstituted and authentic epidermis. (ii) Nuclear extracts from cultures enriched in differentiated keratinocytes display an 80-90% reduction of AP-1 activity when compared to extracts from cultures enriched in nondifferentiated cells. (iii) Cytosolic extracts obtained from cultures enriched in differentiated cells reduce, in a concentration-dependent manner, the AP-1 activity present in nuclear extracts of both mammalian and Drosophila cells. (iv) The specific TRE binding activity of a recombinant c-Jun protein is significantly reduced by cytosolic extracts of differentiated keratinocytes, while the specific DNA binding of the purified recombinant human homeoprotein HOX4B is not. (v) The dephosphorylation, by alkaline phosphatase, of cytosolic extracts increases the inhibitory activity already present or makes evident a latent activity. PMID- 8416792 TI - The retinoblastoma gene product is bound in the nucleus in early G1 phase. AB - The product of the retinoblastoma susceptibility gene (pRB) exerts its growth regulatory effects during the G1 phase of the cell cycle, where all pRB present has been assumed to be in the underphosphorylated form. We demonstrate here that pRB is underphosphorylated and firmly bound in the nucleus only in early G1 phase. All G0 cells contain bound, underphosphorylated pRB. The duration of the cell cycle and of the G1 phase seems to be determined by the time during which pRB is underphosphorylated and bound in the nucleus. The observed time lag between the phosphorylation and release of pRB in the G1 phase and entry into S phase was 6.5 h and independent of the G1 transit time. The data suggest that pRB is not directly involved in initiation of DNA replication. PMID- 8416793 TI - Effect of hydrogen peroxide on cytoskeletal proteins of Drosophila cells: comparison with heat shock and other stresses. AB - Hydrogen peroxide, which was shown to trigger the heat-shock response by activating the immediate binding of the heat-shock factor to DNA heat shock regulatory elements in the promoter of heat-shock genes of Drosophila cells, has also been reported to enhance the synthesis of actin. We show here that very short and transient H2O2 treatments, from 1 s to 2 min, are sufficient to induce an increase of actin synthesis. This increase becomes apparent 2 to 3 h after the short H2O2 treatment. It is inhibited if actinomycin D is present during the short H2O2 treatment. An increase of actin synthesis was also observed during the recovery period after two other stresses: reoxygenation after anoxia and ethanol treatment. The synthesis of two cytoskeletal proteins, tubulin and a 46-kDa insoluble protein of the intermediate filament fraction, was also slightly increased by H2O2 in Drosophila cells, but this increase was not actinomycin D dependent. H2O2 does not provoke the translocation of the 46-kDa protein to the nuclear fraction as does heat shock. The very rapid stimulation of actin synthesis by H2O2 and the involvement of cytoskeletal elements in many stress situations suggest that actin may play a key role in the response to external stimuli. PMID- 8416794 TI - Mechanisms through which gangliosides inhibit PDGF-stimulated mitogenesis in intact Swiss 3T3 cells: receptor tyrosine phosphorylation, intracellular calcium, and receptor binding. AB - Several potential mechanisms through which gangliosides could modulate PDGF stimulated events in Swiss 3T3 cells were studied using intact cells. Of the gangliosides studied, at low micromolar concentrations GM2 was the only one that inhibited PDGF-stimulated DNA synthesis, but GT1b was the most potent between 25 and 100 microM; GM1 was generally the least effective, and GD1a and GM3 had intermediate effects. All gangliosides tested inhibited the PDGF-stimulated increases in free intracellular calcium concentrations ([Ca2+]i) with the rank order of potency being GM1 > or = GT1b > GM2 > GM3. PDGF stimulated phosphorylation on tyrosine of a protein with apparent M(r) = 170 kDa which was immunoprecipitated by an anti-PDGF receptor (beta) antibody, indicating that it is a PDGF receptor. Preincubating the cells with specific gangliosides inhibited tyrosine phosphorylation of this protein in a dose-responsive fashion with the following rank order of potency GD1a = GT1b > GM1 > GM2 > GM3. Autoradiography showed that this was due to a decrease in the proportion of cells synthesizing DNA, and a time study showed that ganglioside did not delay entry of the cells into S phase. These effects were not due to gangliosides interfering with PDGF binding to its receptor because results of competitive binding studies showed that none of the gangliosides studied had an effect on either receptor number or affinity, and did not bind to PDGF in solution. These results show that gangliosides affect several specific components of the complex responses to PDGF in intact cells. The relative effectiveness of individual gangliosides, however, varied among the different cellular and molecular responses. This is interpreted to mean that specific gangliosides modulate to different degrees several molecular mechanisms which converge on the common biological response of mitogenesis, and suggests that gangliosides as a family of molecules may function as coordinators of different molecular events involved in complex cellular processes. PMID- 8416795 TI - Synthesis and expression of smooth muscle phenotype markers in primary culture of rabbit aortic smooth muscle cells: influence of seeding density and media and relation to cell contractility. AB - Rabbit aortic smooth muscle cells (SMC) were seeded at moderate or high densities and grown either in the presence of serum or in the serum-substitution formula Monomed. Expression and synthesis of marker proteins caldesmon, calponin, smooth muscle myosin, and vinculin were monitored during SMC cultivation. Contractility was tested by the ability of cultured SMC to deform silicone membranes following ionomycin treatment. The results show that cells of moderate density grown in Monomed, as opposed to those grown in 5% serum, have the smooth muscle isoform of caldesmon 1.6-fold higher, calponin 1.4-fold and smooth muscle myosin 1.4-fold higher on Day 14 of cultivation. Synthesis of these proteins corresponded to their expression in SMC. The metavinculin:vinculin ratio slightly decreased over the first days with a following reestablishment on Day 8. Contraction was observed until Day 13, compared with Day 7 for cells grown in the presence of serum. High seeding density also prevented a decrease in the expression of smooth muscle markers with the exception of smooth muscle caldesmon whose content in the high density SMC culture was not significantly different from that in the moderate density culture. The period of contractility of SMC in the high density culture was also similar to that in the moderate density culture in the presence of serum. We conclude that cultivation of primary SMC in Monomed allows the maintenance of cells in the contractile phenotype more effectively than high initial seeding density. PMID- 8416796 TI - Activation of apoptosis by serum deprivation in a teratocarcinoma cell line: inhibition by L-acetylcarnitine. AB - P19 teratoma cells differentiate to neural-like cells in the presence of retinoic acid. If they are plated in N2 synthetic, serum-free medium without being exposed to retinoic acid, they die within 48-72 h. This model has allowed the discovery of the neuron survival-promoting capacity of activin. We have studied the death process triggered by serum removal, and showed that it has the characteristics of apoptosis. In addition we have used this model to study the mechanism of action of L-acetyl-carnitine. This endogenous molecule has been successfully employed as a drug retarding Alzheimer's disease progression. Many pharmacological actions have been reported for this compound. However, so far, it has been difficult to explain the observed results with a single mechanism of action. We have demonstrated that the addition of 100 microM L-acetylcarnitine to the N2 medium, at the time of plating, enhances cell survival, retarding DNA fragmentation and nuclear condensation. We have ruled out the possibility of a role of oxidative stress in the activation of apoptosis, under our conditions. Therefore the protective action of L-acetylcarnitine does not seem to be due to a putative antioxidant activity. Our data, demonstrating a retardation by L-acetylcarnitine of apoptotic cell death, could provide a unifying hypothesis for the explanation of several described actions of this drug. In the view that some of the degenerative diseases in the nervous system could be due to the presence of abnormal stimuli, or the absence of trophic factors that trigger programmed cell death, this model of serum deprivation-induced cell death seems to be relevant for the study of neuroprotective molecules. PMID- 8416797 TI - Different subcellular localizations of discoidin I monomer and tetramer in Dictyostelium discoideum cells: using conformation-specific monoclonal antibodies. AB - We characterized four monoclonal antibodies that recognize monomeric and/or tetrameric forms of Dictyostelium lectin (discoidin I) to study intracellular localization of this lectin in early development. Three different PAGEs (native-, urea-, and SDS-PAGE) following immunoblot showed that three of the four mAbs exhibit preference for the tetrameric form while mAb DC2 reacts only with unreduced monomer. By immunofluorescence studies of Dictyostelium NC-4, we found that the anti-tetramer antibodies mainly stain multilamellar bodies, which are food vacuoles to be externalized from cells. In contrast, mAb DC2 stains the cytosol weakly but not the multilamellar bodies. The same pattern of distribution was confirmed by immunoelectron microscopy. These results clearly indicate that, upon packaging discoidin I into multilamellar bodies, the tetrameric form is selectively packaged from the cytosolic pool. In addition, to clarify the relation between the multilamellar bodies and the tetramer, we examined the localization of tetramer using both the cells fed by Escherichia coli and the A3 cells axenically grown. When the cells were fed by E. coli, the cells made multilamellar bodies that contained no lectin; instead the tetramer was found in the cytoplasm and the cells still exclude the lectin around the cells. In contrast, A3 cells grown axenically without bacteria do not make multilamellar bodies but the tetramer also was found in the cytoplasm evenly (also, see W.F. Loomis, Dictyostelium Discoideum, pp. 160-161, Academic Press, New York, 1975). The data suggest that, under these conditions, the tetramer can be excluded from cells via some organelle such as normal secretory vacuole but not multilamellar bodies. Moreover this represents the first example of differential localization of tetramer and monomer of discoidin I in the cellular slime mold cells. PMID- 8416798 TI - Relationship of levels and kinetics of H-ras expression to transformed phenotype and loss of TGF-beta 1-mediated growth regulation in intestinal epithelial cells. AB - Immortalized, nontumorigenic rat intestinal epithelial cells (IEC-18) can be transformed to tumorigenicity by expression of an activated human H-ras gene. Here we describe the characteristics of an IEC-18 cell line in which the activated human H-ras gene has been introduced under the control of the steroid sensitive promoter of the mouse mammary tumor virus long-terminal repeat. The clonal cell line (IEC-18 C125) is phenotypically normal in the absence of the transcription inducer, dexamethasone, and transformed when treated with high levels of inducer. Transformed morphology and growth characteristics are dependent on levels of H-ras expression. IEC-18 C125 cells have been used to demonstrate a general relationship (dose and kinetic) between H-ras expression and loss of TGF-beta 1-mediated growth regulation. This effect occurs concomitantly with a ras-dependent change in the profile of TGF-beta 1 binding proteins. PMID- 8416799 TI - Effect of alpha 1-adrenergic blockade on nucleolar growth, chromatin relaxation, and histone H1(0) content in regenerating liver. AB - alpha 1-Adrenergic agonists are known to be involved in the regulation of hepatocyte proliferation after a partial hepatectomy. The blockade of alpha 1 adrenergic receptors with the specific antagonist prazosin inhibits DNA synthesis which peaks at 24 h after surgery. In this report we have studied the effects of prazosin administration on several events occurring during liver regeneration. The results show that the nuclear volume and nucleolar volume density of hepatocytes were increased and that the relative amount of heterochromatin decreased at 24 h. The increase in hepatocyte nucleolar volume density and the decrease in the relative amount of heterochromatin were partially abolished by prazosin administration while the increase in the nuclear volume was not affected. The relative amount of the histone H1 variant H1(0) was reduced in 24-h regenerating liver and prazosin treatment prevented this reduction. PMID- 8416800 TI - Differentiation-linked expression of prothymosin alpha gene in human myeloid leukemic cells. AB - Prothymosin alpha (ProT alpha) is a nuclear protein related to cell proliferation. Its gene is highly activated during postnatal development at stages containing many proliferating but also differentiating cells. In this report, a study on ProT alpha gene expression during differentiation of human myeloid leukemic (HL-60) cells was undertaken to analyze the possible association of ProT alpha to cell differentiation. When HL-60 cells were induced to differentiate to granulocytes (using retinoic acid) or monocyte/macrophages (using 12-O-tetradecanoylphorbol-13-acetate), a marked down-regulation in the levels of ProT alpha transcript was found. When cell division of immature HL-60 cells was interrupted by either treatment with hydroxyurea or serum starvation, ProT alpha gene expression was not significantly altered. These findings suggest that loss of ProT alpha mRNA in induced HL-60 cells is a differentiation-related event. Examination of the stability of ProT alpha mRNA showed that the stabilization of the ProT alpha transcript is differentially regulated in the two HL-60 lineages. Nuclear run-on experiments revealed that during HL-60 differentiation, the transcriptional activity of the ProT alpha gene does not experience significant variations. PMID- 8416801 TI - Quantitative determination of oxidative base damage in DNA by stable isotope dilution mass spectrometry. AB - For understanding of the role of oxidative DNA damage in biological processes such as mutagenesis and carcinogenesis, it is essential to identify and quantify this type of DNA damage in cells. This can be achieved by gas chromatography/mass spectrometry. The present study describes the quantification of modified bases in DNA by isotope-dilution mass spectrometry with the use of stable isotope-labeled analogues as internal standards. A number of isotopically labeled DNA bases were synthesized. The mass spectra of their trimethylsilyl derivatives were recorded. Calibration plots were obtained for known quantities of modified bases and their isotope-labeled analogues. Quantification of various modified DNA bases by isotope-dilution mass spectrometry was demonstrated in isolated chromatin exposed to ionizing radiation. The results indicate that gas chromatography/stable isotope-dilution mass spectrometry is an ideally suited technique for selective and sensitive quantification of modified bases in DNA. PMID- 8416802 TI - Bibrotoxin, a novel member of the endothelin/sarafotoxin peptide family, from the venom of the burrowing asp Atractaspis bibroni. AB - A new member of the endothelin/sarafotoxin family of vasoconstrictor peptides, bibrotoxin (BTX), was isolated from the venom of the burrowing asp Atractaspis bibroni by reversed-phase FPLC. The amino acid sequence of BTX differs from SRTX b in the substitution Ala4 instead of Lys4, which suggests that it represents the peptide isoform of Atractaspis bibroni corresponding to SRTX-b. BTX competed for [125I]ET-1 binding to human ETB-type receptor with a Ki of 3.2 x 10(-9) M compared to 4.2 x 10(-9) M for SRTX-b. In rat thorax aorta BTX induced vasoconstrictions with a threshold concentration of 3 x 10(-8) M compared to 1 x 10(-9) for ET-1. PMID- 8416803 TI - The NMR determination of the IIA(mtl) binding site on HPr of the Escherichia coli phosphoenol pyruvate-dependent phosphotransferase system. AB - The region of the surface of the histidine-containing protein (HPr) which interacts with the A domain of the mannitol-specific Enzyme II (II(Amt1)) has been mapped by titrating the A-domain into a solution of 15N-labeled HPr and monitoring the effects on the amide proton and nitrogen chemical shifts via heteronuclear single quantum correlation spectroscopy (HSQC). Fourteen of the eighty-five HPr amino acid residues show large changes in either the 15N or 1H chemical shifts or both as a result of the presence of II(Amt1) while a further seventeen residues experience lesser shifts. Most of the residues involved are surface residues accounting for approximately 25% of the surface of HPr. Phosphorylation of HPr with catalytic amounts of Enzyme I (EI), in the absence of II(Amt1) resulted in chemical shift changes in a sub-set of the above residues; these were located more in the vicinity of the active site phospho-histidine. Phosphorylation of the HPr/II(Amt1) complex resulted in a HSQC spectrum which was indistinguishable from the P-HPr spectrum in the absence of II(Amt1) indicating that, as expected, the complex P-HPr/P-II(Amt1) does not exist even at the high concentrations necessary for NMR. PMID- 8416804 TI - Selective cleavage of 28S rRNA variable regions V3 and V13 in myeloid leukemia cell apoptosis. AB - Vigorous apoptosis is induced 3-4 hours after activation of cAMP-dependent protein kinase I in the rat myeloid leukemia cell line IPC-81 [J. Cell. Physiol. 146 (1991) 73-80]. We will report a novel feature of apoptosis in these cells: a selective and temporarily ordered cleavage within the two largest 28S ribosomal RNA variable regions (V3 and V13). The cleavage of 28S rRNA coincided with internucleosomal DNA fragmentation and cessation of cellular protein synthesis. The implication of 28S variable regions as targets in apoptosis is a clue to the function of these so far apparently superfluous parts of eukaryotic ribosomes. PMID- 8416805 TI - Nitric oxide preferentially stimulates auto-ADP-ribosylation of glyceraldehyde-3 phosphate dehydrogenase compared to alcohol or lactate dehydrogenase. AB - Recently we demonstrated that the radical nitric oxide (NO) stimulates the auto ADP-ribosylation of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) resulting in enzyme inhibition. To further characterize this auto-ADP-ribosylation reaction we studied alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH) for comparison. Whereas auto-ADP-ribosylation of ADH was stimulated to a minor extent by the NO-liberating agent 3 morpholinosydnonimine (SIN-1), LDH was unaffected. The susceptibility of dehydrogenases towards auto-ADP-ribosylation correlated with the potency of NO to decrease enzyme activity. Again, GAPDH was much more sensitive compared to ADH, whereas LDH again was unaffected. Interestingly, the efficiency of the SH alkylating agent N-ethylmaleimide (NEM) to inhibit the enzymatic activity of the chosen dehydrogenases correlates with the sensitivity of dehydrogenases towards NO. These studies demonstrate the requirement of a reactive SH-group besides the NAD+ binding site as a prerequisite for NO-stimulated auto-ADP-ribosylation reactions. Furthermore, we establish that under physiological conditions and among the dehydrogenases tested, only GAPDH is a potential target for this post translational protein modification mechanism. PMID- 8416806 TI - Transfer of foreign DNA into the cells of developing mouse embryos by microprojectile bombardment. AB - Mouse cells of developing embryos at the 2-4 cell, morula and blastocyst stages, were bombarded by high velocity tungsten microprojectiles. About 70% of developing embryos survived the bombardment. The general embryo structure did not change as a result of the bombardment. Penetration of the tungsten microparticles into the embryo cell nuclei was found at all stages being investigated, and tungsten particle localization on mitotic chromosomes was demonstrated. The total DNA of the mice born from the bombarded embryos was analyzed by dot-blot hybridization and PCR with post-hybridization. The most important results were obtained in experiments with blastocysts. In three cases of blastocyst bombardment, the presence of transferred plasmid DNA (pSV3-neo) was revealed. Transfected cells were shown to be located in the fetal membrane as well as in the embryo. The bombardment of mouse culture cells resulted in their transfection and the production of G418-resistant clones. PMID- 8416807 TI - Staurosporine stimulates phospholipase D activation in human polymorphonuclear leukocytes. AB - Treatment of 1-O-[3H]alkyl-2-acyl-phosphatidylcholine-prelabeled human polymorphonuclear leukocytes (PMNs) with staurosporine (50 nM to 1 microM) induced a time- and concentration-dependent generation of tritiated phosphatidic acid (PA), reaching approximately 225% of the control value at 15-20 min. In the presence of ethanol, staurosporine induced a production of phosphatidylethanol (PEt) reaching, 250% of control values, and partial inhibition of PA production, consistent with PLD activation. The amount of ether-linked acylglycerol (EAG) was weakly enhanced (29%) after 5 min of PMN treatment; longer treatment resulted in no significant EAG production, suggesting a possible late inhibition of PA hydrolase activity. Staurosporine concentrations that induced an elevation in PA completely depressed protein kinase C (PKC) activity in both soluble and particulate cell fractions, suggesting that PLD activation may occur independently from PKC activation. PLD may thus represent a potential cellular target for staurosporine action. PMID- 8416808 TI - X-ray crystallographic structure of a papain-leupeptin complex. AB - The three-dimensional structure of the papain-leupeptin complex has been determined by X-ray crystallography to a resolution of 2.1 A (overall R-factor = 19.8%). The structure indicates that: (i) leupeptin contacts the S subsites of the papain active site and not the S' subsites; (ii) the 'carbonyl' carbon atom of the inhibitor is covalently bound by the Cys-25 sulphur atom of papain and is tetrahedrally coordinated; (iii) the 'carbonyl' oxygen atom of the inhibitor faces the oxyanion hole and makes hydrogen bond contacts with Gln-19 and Cys-25. PMID- 8416809 TI - Effect of carnitine feeding on the levels of heart and skeletal muscle carnitine of elderly mice. AB - Aging has been associated with an increase in muscle dysfunction and weakness. We found a decrease in muscle carnitine with age [Biochem. Biophys. Res. Commun., 161 (1989) 1135-1143]. Prolonged oral administration to both young (2-month-old) and adult (7-month-old) mice with L-carnitine increased its content in blood by 50%. The levels of carnitine in skeletal and heart muscle of old treated animals became higher than in untreated mice of the same age. However, this extensive restoration did not reach the maximum values present in skeletal muscle of young mice. Our findings indicate that an alteration of the carnitine carrier in the sarcolemma could be responsible for the decrease with age of carnitine in skeletal but not in heart muscle. PMID- 8416810 TI - Effects of beta 6 amino acid hydrophobicity on stability and solubility of hemoglobin tetramers. AB - The relationship between different amino acids at the beta 6 position of hemoglobin and tetramer stability was addressed by a site-directed mutagenesis approach. Precipitation rates during mechanical agitation of oxyhemoglobins with Gln, Ala, Val, Leu and Trp at the beta 6 position increased 2, 5, 13, 21 and 53 times, respectively, compared with that for Hb A. There was a linear relationship between the log of the precipitation rate constant and amino acid hydrophobicity at the beta 6 position, suggesting that enhanced precipitation of oxy Hb S during mechanical agitation results in part from increased hydrophobicity of beta 6 Val. Deoxyhemoglobin solubility increased in the order of beta 6 Ile, Leu, Val, Trp, Gln, Ala and Glu suggesting that hydrophobic interactions between beta 6 Val and the acceptor site of another hemoglobin molecule during deoxy-Hb S polymerization not only depend on hydrophobicity but also on stereospecificity of the amino acid side chain at the beta 6 position. Furthermore, our results indicate that hydrophobic amino acids at the beta 6 position which promote tetramer instability in the oxy form do not necessarily promote polymerization in the deoxy form. PMID- 8416811 TI - The DNA-binding properties of an artificial 42-residue polypeptide derived from a natural repressor. AB - Bacteriophage 434 repressor recognizes the operator sequences ACAAG and ACAAT. As the same or similar sequences occur in the enhancer region of HIV-1, 434 repressor was a potential HIV enhancer-binding protein. We found that the interaction of the DNA-binding domain of 434 repressor with a 57-bp HIV enhancer DNA was very weak whereas a 42-residue construct, comprising the recognition helix and four copies of a positively charged segment of the repressor, bound strongly. The results of footprint and cell-free in vitro transcription studies showed that the 42-residue peptide bound preferably to the enhancer region of HIV 1 and acted as an artificial repressor. Replacement of an essential glutamine of the recognition helix by glutamic acid resulted in a partial shift of the sequence specificity of the 42-residue peptide. PMID- 8416812 TI - Methyl green. A DNA major-groove binding drug. AB - Interaction and binding geometries of complexes of Methyl green with poly(dA dT)2, poly(dA).poly(dT), and triplex poly(dA).2poly(dT) complexes have been studied by linear dichroism. For both of the complexes with double helical DNAs, the z symmetry axis of Methyl green is found to be approximately parallel to the DNA bases while the x symmetry axis lies at 40-44 degrees relative to the local DNA helix axis, in agreement with a groove binding mode. However, in contrast to minor-groove binders (such as DAPI and Hoechst 33258) Methyl green is found to be excluded from binding to the triple helical poly(dA).2poly(dT) in which the major groove is filled by the third strand. While most so far studied groove-binding dyes bind in the minor groove of DNA, Methyl green thus appears to be an exception. PMID- 8416813 TI - Multiple UDP-glucuronyltransferases for the glucuronidation of thyroid hormone with preference for 3,3',5'-triiodothyronine (reverse T3). AB - We have studied the glucuronidation of the thyroid hormones T4, T3 and rT3 by liver microsomes of Wistar, Gunn and WAG rats. Gunn rats have a defect in the gene coding for bilirubin and phenol UDP-glucuronyltransferase (UGT) isoenzymes; WAG rats have a genetic defect in androsterone UGT. In normal Wistar rats UGT activity was approximately 5-fold higher for rT3 than for T4 or T3. UGT activities for T4 and rT3, but not for T3, were impaired in Gunn rats. Conversely, UGT activity for T3, but not for T4 or rT3, was impaired in WAG rats. Thus, in rat liver rT3 is glucuronidated much more rapidly than T4 and T3. Our results support the view that T4 and rT3 are glucuronidated by bilirubin and phenol UGTs and T3 by androsterone UGT. PMID- 8416814 TI - Biochemical evidence that phytochrome of the moss Ceratodon purpureus is a light regulated protein kinase. AB - The phytochrome gene of the moss Ceratodon purpureus (phyCer) codes for a novel phytochrome polypeptide with a predicted molecular mass of 145 kDa that has a COOH-terminal domain which is homologous to the catalytic domain of eukaryotic protein kinases. In this paper we report the first biochemical evidence that in fact, as predicted from the gene sequence, PhyCer represents an active, light regulated protein kinase. In vitro phosphorylation experiments with protonemata extracts revealed the existence of a 140 kDa protein, phosphorylated in a red/far red light dependent manner. The binding of a polyclonal antibody directed to the protein kinase catalytic domain of PhyCer enhanced the phosphorylation of a 140 kDa band when assayed in a renaturation-auto-phosphorylation experiment with nitrocellulose bound protein. These findings strongly implicate that the phyCer gene product has protein kinase activity and is capable of auto-phosphorylation. The results of the renaturation-phosphorylation experiments were essentially the same, no matter whether protein extracts from light grown or dark adapted moss protonemata were used. Thus, phyCer expression most likely is not light regulated. PMID- 8416815 TI - Identification of toxigenic Vibrio cholerae from the Argentine outbreak by PCR for ctx A1 and ctx A2-B. AB - A polymerase chain reaction (PCR) to detect a region of the A1 cholera toxin gene was applied to the identification of 43 Vibrio cholerae strains isolated from the recent outbreak in Argentina. A good correlation was observed between the GM1 enzyme-linked immunosorbent assay (GM1-ELISA) to detect the B subunit of the enterotoxin and PCR. However, a V. cholerae non-01 strain that was negative by the ELISA test, was positive by the PCR assay for the A1 region. A second PCR test to detect the A2-B coding region was developed to solve this case. We propose that routine detection of toxigenic V. cholerae by PCR should include analysis of A2-B coding region or the whole cholera toxin operon. PMID- 8416816 TI - Human liver microsomal glutathione transferase. Substrate specificity and important protein sites. AB - Human liver microsomal glutathione transferase displays the following glutathione peroxidase/transferase activities: dilinoleoylphosphatidylcholine hydroperoxide (0.03 and 0.17 mumol/min.mg, unactivated and N-ethylmaleimide-activated enzyme, respectively), linoleic acid hydroperoxide (0.09 and 0.15 mumol/min.mg), cumene hydroperoxide (0.04 and 3 mumol/min.mg), methyl linoleate ozonide (0.02 and 1.2 mumol/min.mg) and 1-chloro-2,4-dinitrobenzene (1.9 and 24 mumol/min.mg). The activation of glutathione peroxidase activities are much higher than previously observed. The activity towards a phospholipid hydroperoxide is noteworthy since protection against lipid peroxidation has been implied. Methyl linoleate ozonide has not previously been characterised as substrate for any microsomal glutathione transferase. Human liver microsomal glutathione transferase displays an isoelectric point of 9.4 and a structure in agreement with that deduced from the cDNA sequence. Gel electrophoretic analysis shows that proteolytic activation of the human enzyme corresponds to cleavage at Lys-41, thus defining the critical activation site. PMID- 8416817 TI - Continuous presence of phorbol ester is required for its IL-1 beta mRNA stabilizing effect. AB - The protein kinase C (PKC) activating phorbol esters are known to prevent the decay of mRNA of several cytokines and proto-oncogenes. To examine whether the phorbol ester signal is continuously required for this stabilizing effect, THP-1 monocytic cells were stimulated either with phorbol 12,13-dibutyrate (PDBu), which can be removed from the cells by washings, or with the more hydrophobic phorbol 12-myristate 13-acetate (PMA). Both of these stimuli induced high levels of interleukin-1 beta (IL-1 beta) mRNA. When the cells were washed at the peak of the IL-1 beta mRNA expression, this mRNA decayed rapidly in the PDBu stimulated cells while in PMA stimulated cells the mRNA levels were not affected. Moreover, this mRNA degradation induced by the removal of PDBu could be inhibited by readdition of the phorbol ester. This restabilization could be prevented by pharmacologic inhibitors of PKC, but not by inhibiting protein or RNA synthesis. Thus these data suggest that the phorbol ester must be continuously present to exert its mRNA stabilizing effect and that its effect is PKC-mediated but does not require active protein or RNA synthesis. PMID- 8416818 TI - Disulfide bridges in human complement component C3b. AB - The disulfide bridges of human complement component C3b, derived from C3 by removal of the 77-residue C3a, have been determined. The 10 bridges are Cys537 Cys794, Cys605-Cys640, Cys851-Cys1491, Cys1079-Cys1136, Cys1336-Cys1467, Cys1367 Cys1436, Cys1484-Cys1489, Cys1496-Cys1568, Cys1515-Cys1639, and Cys1615-Cys1624. Including the 3 bridges in C3a (Cys670-Cys698, Cys672-Cys705, and Cys685-Cys706) previously determined by high-resolution X-ray crystallography [Hoppe-Seyler's Z. Physiol. Chem. 361 (1980) 1389-1399] all disulfide bridges of C3 are localized. C3 and the strongly related C4 and C5 are members of the alpha 2-macroglobulin superfamily. The predicted bridge patterns of C4 and C5 are discussed and compared with that of alpha 2-macroglobulin. PMID- 8416819 TI - VAT-1 from Torpedo is a membranous homologue of zeta crystallin. AB - VAT-1 is a major protein from Torpedo synaptic vesicles. A protein data-base search revealed a striking homology to zeta crystallin from guinea pig lens. The overall amino-acid identity is 27%, and 58% similarity is reached by including conserved substitutions. The highest similarity (60% to 85%) between the two proteins is observed in five discrete domains, which are also conserved in zinc dependent dehydrogenases, particularly in the alcohol dehydrogenase family. The cofactor-binding domain of oxidoreductases is conserved in VAT-1 and in zeta crystallin. VAT-1 preferably binds NADPH in the presence of zinc. In contrast with its homologous proteins, VAT-1 is an integral membrane protein of synaptic vesicles. PMID- 8416820 TI - The basic principles of infectious diseases as related to dental practice. AB - A variety of bacterial, viral, fungal, and protozoan microbes may present hazards to the dental team and to patients. After contamination with a microbe, three major factors determine if an infectious disease results: (1) virulence of the microbe, (2) the dose or number of microbial cells that contaminate the body, and (3) the resistance of the body to the microbe. Because nothing can be done about microbial virulence, prevention of infectious diseases involves decreasing the dose and increasing body resistance, coupled with education about disease spread and modes of prevention. The steps in the development of an infectious disease are (1) source of the microbe, (2) escape of the microbe from the source, (3) spread of the microbe to a new person, (4) entry of the microbe into the new person, and (5) infection and damage. The prevention of infectious disease can be accomplished by interfering with any of the above steps. PMID- 8416821 TI - Computer applications in oral diagnosis. AB - Eventually, computer technology may be used in the home or practice to satisfy continuing education requirements. Additionally, this technology may be used by state licensing agencies as a component of their testing procedure. At the present time, a Patient Simulation Consortium and Electronic Curriculum Consortium have been established within the American Association of Dental Schools.7, 15 These groups, and others, will continue to develop and test new technologies for use in dentistry. PMID- 8416822 TI - Forensic and legal issues in oral diagnosis. AB - The forensic odontologist has assumed a more visible role in the last decade, having been called upon repeatedly to assist both law enforcement agencies and the judicial system. It has become quite common for dentists to establish the identity of unknown bodies; to quickly and positively identify the victims of mass disasters such as airliner crashes, floods, and earthquakes; as well as to provide testimony in court concerning bite marks or other matters that require dental expertise. Litigation directed at dental defendants for failure to diagnose, properly treat, or obtain valid consent from plaintiff patients is also on the rise. In each of these fields, dentists who have been specially trained in or who have devoted their practices to the dental diagnostic sciences have been looked to widely as experts. This article discusses in detail these areas and others that affect the dental diagnostician. PMID- 8416823 TI - Oral manifestations of human immunodeficiency virus infection. AB - Oral lesions were included in some of the first descriptions of the acquired immunodeficiency syndrome (AIDS), and it has become clear in the ensuing years that these lesions may represent some of the earliest signs of infection with the human immunodeficiency virus (HIV), may be of prognostic significance in the subsequent development of AIDS, and may, if not treated, produce significant morbidity. Understanding oral lesions may shed light on the other manifestations of HIV disease. Oral examination is an important part of any physical examination, and nowhere is this more important than in the case of suspected HIV infection. PMID- 8416824 TI - Diagnosis and management of taste disorders and burning mouth syndrome. AB - Clinically significant taste loss is less common than abnormal tastes (dysgeusias). Both may be caused by a previous viral upper respiratory infection, head trauma, iatrogenic causation (medication, irradiation, surgery), neurologic or psychiatric disorders, toxic chemical exposure, systemic conditions, xerostomia, severe nutritional deficiencies, and some oral or dental disorders. Beyond treatment targeted toward causative conditions, there is no proven intervention to either enhance taste acuity or abolish dysgeusia. The prevalence of oral burning sensations has been estimated at 2.6% for the general population. The burning typically increases throughout the day, and may be associated with taste alterations and psychological effects. Differential diagnoses considered include psychiatric illness, menopause, nutritional disorders, oral and dental conditions, and diabetes mellitus. Low doses of tricyclic antidepressants may be effective in some patients with idiopathic oral burning, and spontaneous remissions without intervention have been reported. PMID- 8416825 TI - Clinical implications of drugs taken by our patients. AB - An awareness of the various medications commonly prescribed for patients to self administer will assist clinicians to anticipate the most commonly encountered medical diagnoses; will give clues to a patient's physical and emotional ability to undergo and respond to dental care; will alert clinicians to potential drug drug and drug-disease interactions, and to the presence of drug-induced illness; and provide invaluable information that will help the clinician identify high risk patients who may experience a life-threatening medical emergency while in the dental office. PMID- 8416826 TI - Advances in diagnostic imaging in dentistry. AB - Advances in optical and x-ray imaging will affect dramatically the practice of dentistry in the next decade. "Hard copy" of these images will be on thermographic paper from videoprinters as well as x-ray film. Reconstruction of images in both (two and three dimensions) will allow better visualization of disease processes and allow quantification of disease changes over time, thus assisting the treatment planning decisions of the dentist and improving patient care. Some of these innovations are here, others are in the research and development stage. All dentists eventually will use these techniques routinely. PMID- 8416827 TI - Different members of the fibroblast growth factor receptor family are specific to distinct cell types in the developing chicken embryo. AB - Single-stranded RNA probes for the three chicken fibroblast growth factor (FGF) receptors, cek-1, cek-2, and cek-3, in conjunction with in situ hybridization were used to characterize the distribution of the corresponding mRNAs in the developing chicken embryo. Cek-1 was expressed diffusely in most tissues examined, whereas the expression of cek-2 and cek-3 was more restricted. The highest levels of FGF receptor expression were seen in the developing bones; in skeletal, cardiac, and smooth muscle; and in some areas of the brain. Although all three receptors were expressed in a number of the same tissues, the expression of each receptor within a given tissue was generally specific for different cell types. In addition, the distribution of each of these receptors did not correlate with the previously characterized distributions of individual FGFs. These results suggest that the members of the FGF receptor family may represent cell-type-specific receptors rather than ligand-specific receptors. Thus, the interaction between a growth factor of the FGF family and a given FGF receptor is likely to be controlled to a large extent by spatial constraints, rather than exclusively by high binding affinities. PMID- 8416828 TI - Biogenesis of surface domains during spermiogenesis in the guinea pig. AB - During mammalian spermiogenesis a spherical spermatid is transformed into a highly asymmetric sperm cell. Concurrently, the plasma membrane of the cell develops into a mosaic of discrete membrane regions, with each region containing a unique set of proteins. Biogenesis of these surface domains was studied by following the surface expression and localization of nine different antigens during spermiogenesis. Each of these antigens exhibits one of four distinct patterns of localization on testicular sperm (whole cell, whole head, anterior tail, and posterior tail), indicating that there are at least three distinct surface domains on testicular sperm. Our results on the timing of antigen localization suggest that the generation of surface domains in mammalian sperm is a complex process. This process involves temporal and spatial regulation of surface expression of the antigens, as well as the specific removal of antigens from inappropriate domains after they have reached the cell surface. PMID- 8416829 TI - Morphological and physiological development of vestibular hair cells in the organ cultured otocyst of the chick. AB - The inner ear of the embryonic chick forms an oval-shaped sac or otocyst, on Embryonic Day 3, which contains presumptive sensory and support cells. After 3 weeks in organ culture the otocyst had sensory epithelia with an average of 325 +/- 41 hair cells. Using light and transmission electron microscopy most of these cells were identified morphologically as type II vestibular hair cells. Whole cell tight-seal recordings, using potassium chloride-filled micropipetes, showed that mature cultured hair cells had four different types of K+ currents. These included: a voltage-gated delayed rectifier K+ current (IK), an inactivating K+ current (IA), a calcium-dependent K+ current (IK(Ca)), and a K+ inward rectifier (IIR). These currents were similar to those recorded from cristae ampullares cells isolated from 2- to 3-week-old posthatched chicks. We also determined the timing of K+ current acquisition in vitro. Initially, recordings showed that cells isolated from Embryonic Day 3 otocysts had no voltage-dependent outward currents at physiological membrane potentials. Eventually, K+ currents were acquired in the order of: IK and IIR after 9 days, IA after 12 days, and IK(Ca) after 17 days in vitro. In addition, recordings using cesium chloride-filled micropipetes showed that there were two types of inward currents that were elicited in response to membrane depolarizations. These two currents included a rapidly activating, noninactivating Ca2+ current and a tetrodotoxin-sensitive Na+ current. Both currents were elicited in hair cells grown in vitro for 13 days. Although INa was previously unreported in avians, both INa and ICa were also represented in hair cells isolated from the cristae ampullares of the posthatched chick. These results indicate that hair cells can acquire voltage-gated currents in vitro which are qualitatively similar to ionic currents found in crista ampullaris cells that differentiate in vivo. Thus, this organ culture system provides a means to study regulation of ionic currents in developing hair cells. PMID- 8416830 TI - t haplotypes in the mouse compromise sperm flagellar function. AB - The t haplotypes are variant forms of the proximal portion of chromosome 17 in the mouse. The t haplotypes alter spermatogenesis and many also contain lethal factors. Although the lethal factors vary between t haplotypes, all t haplotypes have the same effect on sperm, that of altering sperm function in fertilization. It is not clear, however, whether the nature of the sperm dysfunction is the same in all t haplotypes. Studies to date have focused on only one or two aspects of sperm function or have not examined sperm from males carrying different t haplotypes. In addition, factors within the t haplotypes interact with the genetic background, so that comparisons to sperm from t/+ or +/+ mice having different alleles at loci outside the t haplotypes may not be valid. To determine the nature of the sperm dysfunction caused by the t haplotypes, we have studied sperm from mice of the same genetic strain carrying none, one, or two t haplotypes. Sperm from tw32/+, tw5/+, and tw32/tw5 mice exhibited premature hyperactivation, a type of vigorous but nonprogressive motility correlated with fertility, while their rates of capacitation (the ability to undergo the acrosome reaction in response to zona proteins) and spontaneous acrosome reaction were similar to those of wild-type sperm. In addition, sperm flagellar curvature was abnormal: flagella from heterozygotes had an acute bend in the midpiece, giving the sperm a "fishhook" appearance, while the entire flagellum of sperm from tw32/tw5 mice was curled. Also, fewer sperm from tw32/tw5 mice were initially motile. Since all of these motility defects were dependent on exogenous calcium, the t haplotypes could specify an abnormal, calcium-sensitive component of the flagellum. The motility defects could also contribute to the dysfunction of these sperm by inhibiting their passage to the site of fertilization in vivo. PMID- 8416831 TI - Transient, localized accumulation of alpha-spectrin during sea urchin morphogenesis. AB - The mRNA and protein of alpha-spectrin in the sea urchin embryo is shown here to be transiently overexpressed in cells initiating certain morphogenetic changes. This expression was detected by use of a 4.5-kb cDNA clone that encodes alpha spectrin isolated from a Lytechinus variegatus cDNA library. DNA sequence analysis demonstrated 80% similarity of this cDNA to human nonerythroid alpha spectrin. Sea urchin alpha-spectrin RNA accumulated to a uniform basal level in all cells of the embryo with a marked, transient increased accumulation in the primary mesenchyme cells of the vegetal plate just prior to gastrulation. Cells of the endoderm also express increased levels of spectrin during gastrulation and these levels remained high in the myoepithelial cells of the sphincter constrictions that separates the gut into foregut, midgut, and hindgut domains. Antibodies made to recombinant sea urchin spectrin synthesized in Escherichia coli were used to localize the protein in situ. These experiments showed that all cells of the embryo accumulated alpha-spectrin protein confined to the cortical regions of cell-cell contact. In epithelial cells, this localization showed a distinct honeycomb pattern. Primary mesenchyme cells and the myoepithelial cells of the gut sphincters showed significantly enhanced spectrin signal corresponding to the pronounced RNA localization pattern seen by in situ RNA hybridization. These data demonstrated that alpha-spectrin from the sea urchin is a highly conserved member of the spectrin superfamily of proteins and that its elevated expression in specific cell types anticipates overt morphogenesis. PMID- 8416832 TI - Alpha 5 integrin is a critical component of adhesion plaques in myogenesis. AB - We investigated the distribution and expression of alpha 5 beta 1 and alpha 3 beta 1 integrin in differentiating myogenic cells in culture. The myogenic cells expressed both alpha 5 and alpha 3 integrin subunits with the same molecular sizes as those expressed by chicken embryo fibroblasts (CEF). However, the ratio of total alpha 5 to alpha 3 was threefold higher in the muscle cultures than that in CEF cultures. A new method is described whereby adhesion plaque-associated integrin was cross-linked to its extracellular matrix ligand on the substrate using a nonpenetrating cross-linker, BS3, and integrin not involved in substrate adhesion as well as cytoskeletal proteins were removed with a zwitterionic detergent. alpha 5 and beta 1 integrin, but not alpha 3 could be cross-linked to fibronectin at adhesion plaques throughout myogenesis in culture. Alpha 5 beta 1 integrin was found only at the edge of myoblasts 4 hr after plating but became distributed under their entire surface by 1 day in culture. When the muscle cells became elongated, a morphology they express after the initiation of terminal differentiation, and as they began to fuse, alpha 5 was found redistributed in small adhesion plaques along the lateral edges of the postmitotic myocytes and early myotubes. In mature myotubes, which are large multinucleated branched structures, alpha 5 beta 1 integrin was localized to parallel streaks underneath their entire substrate surface. Throughout the different stages of myogenesis, vinculin colocalized with alpha 5 beta 1 integrin in adhesion plaques, but alpha actinin only colocalized to the adhesion plaques in myoblasts, not in myotubes. These studies suggest that alpha 5 beta 1 integrin through its dynamic interaction with both fibronectin and the cytoskeleton is important for both the signals which initiate the differentiation process and for subsequent morphological and structural changes during the differentiation process. PMID- 8416833 TI - Costimulation-induced rounding in Tetrahymena thermophila: early cell shape transformation induced by sexual cell-to-cell collisions between complementary mating types. AB - Mixing of starved cells of complementary mating types of Tetrahymena thermophila induces shortening of their longitudinal length within 10 min of mixing. This early morphogenetic transformation in preconjugant sexual interaction (costimulation period) was named "costimulation-induced rounding" (CIR). CIR is the earliest morphological change that has ever been found in the costimulation period and differs from "synchronous rounding" in the vegetative cell cycle, because CIR cells are still able to form food vacuoles, while cells in synchronous rounding do not have this ability. When sexual cell-to-cell collisions between two mating types were hampered by unidirectional stirring for 20 min after mixing of the two mating types, both CIR and conjugation were delayed by 20 min. When secreted materials needed for the onset of costimulation were removed by washing the cells with 10 mM Tris-HCl, pH 7.4, before mixing the two mating types, both CIR and conjugation were delayed by about 30 min. CIR-like rounding was not induced by cell-free medium either from the opposite mating type or from mixed costimulated cells. These results indicated that CIR is induced when cells are activated to form conjugating pairs by cell-to-cell collisions between complementary mating types in the presence of secreted molecules. PMID- 8416834 TI - Reevaluation of electrophoresis in the Drosophila egg chamber. AB - To evaluate the hypothesis of electrophoretic transport of cytoplasmic components, the transfollicle potentials of Drosophila oocytes and nurse cells were measured using improved techniques. We found input resistances 20 to 1000 times higher than those in previous reports. Measurements were made in a large variety of conditions: in external potassium concentrations from 1 to 100 mM, over the concomitant membrane potential range -84 to -23 mV, from developmental stages 5 to 10, and with or without using hemolymph, anesthetics, or collagenase. In all of these circumstances, no voltage gradient was detectable with intracellular microelectrodes from nurse cells to oocyte or between nurse cells. No voltage gradient was detected with external suction electrodes. Our results do not support the electrophoretic theory. PMID- 8416835 TI - Major temporal and spatial patterns of gene expression during differentiation of the sea urchin embryo. AB - We have investigated the temporal and spatial patterns of accumulation of mRNAs randomly selected from the sea urchin gastrula polyadenylated RNA population. Three different assays show that the predominant temporal pattern of expression, exhibited by about three-fourths of these messages, consists of a large (mean 80 fold) increase in mRNA abundance between egg and gastrula stages. Most mRNAs are present in the maternal population and are detectable on blots as single mature sized messages; however, a large number of high-molecular-weight, heterodisperse transcripts containing these same sequences also exist in the egg cytoplasm. The majority of gastrula messages are not embryo specific but are present in total adult urchin RNA at concentrations similar to those in embryos. Fine-scale RNA blot analysis indicates that the majority of mRNAs begin to accumulate at very early blastula stages, although there is considerable diversity in the time when these messages reach peak abundance. Most gastrula mRNAs are also spatially regulated during development. The observed distributions can be categorized into three major functional or regulatory classes: (1) Forty percent of mRNAs accumulate in cells which are cycling or preparing for growth. (2) About one third of the messages accumulate in one or more differentiating cell types. (3) Only slightly more than one-fourth of the messages are present in all cell types throughout development. Most tissue-specific messages are relatively abundant, indicating that the differentiated functions of cells are executed through mRNAs operating at the level of hundreds of copies per cell. In contrast, most rare messages are expressed in most or all cell types, in which they function at only a few copies per cell. All messages which begin to accumulate before hatching blastula stage are initially distributed broadly, and their distribution becomes progressively restricted during embryogenesis. In contrast, all messages which begin to accumulate after the onset of gastrulation accumulate only in discrete subsets of cells. The results presented here illustrate much more extensive temporal regulation of gene expression during sea urchin embryogenesis than previously detected. This is accompanied by spatial regulation of expression of most genes which is itself temporally modulated as the cellular requirements for cell division and differentiation change during development. PMID- 8416836 TI - A positive role in differentiation for the male doublesex protein of Drosophila. AB - The doublesex (dsx) locus encodes male-specific and female-specific polypeptides that are essential for the proper differentiation of sexually dimorphic somatic features of Drosophila melanogaster. Ectopic expression of the male-specific dsx polypeptide was obtained by P-element-mediated transformation of flies with a construct bearing a fusion between the hsp70 heat shock promoter and dsx male specific cDNA sequences. Heat shock-induced expression of the male cDNA in either sex resulted in three novel phenotypes: transformation of bristles on all legs toward a sex comb-like morphology, pigmentation of dorsal spinules and ventral setae in third-instar larvae, and lethality. These results were not predicted by previous models of dsx function, and provide evidence that the role of the male dsx protein includes activation of some aspects of male differentiation as well as repression of female differentiation. PMID- 8416837 TI - Promiscuous germination and growth of wildtype pollen from Arabidopsis and related species on the shoot of the Arabidopsis mutant, fiddlehead. AB - When pollen lands upon the stigmatic surface of a receptive flower, recognition events take place that permit germination, pollen tube growth, and tube penetration into the cell walls of the stigmatic papillae. Previously, we have described a mutant of Arabidopsis thaliana, named fiddlehead (fdh), where noncarpel organs of the shoot fuse late in ontogeny (Lolle et al., 1992). Here we demonstrate that wildtype Arabidopsis pollen grows on noncarpel organs of the fdh mutant. Pollen grains adhere, germinate, and emit pollen tubes when applied to vegetative and nonreproductive floral organs. Some of the emergent pollen tubes penetrate into the cell wall. Although pollen from a number of closely related species responds, tomato, tobacco, and snapdragon pollen does not. In addition, we show that organ fusion is not a prerequisite for pollen growth and that root epidermis does not express this activity. Based on these findings we propose that the fdh mutation identifies an important regulatory gene that controls the expression of an epidermis-specific developmental program normally expressed only during gynoecial ontogeny. PMID- 8416838 TI - Origin and development of adventitious shoot meristems initiated on plant chimeras. AB - Most studies concerning the initiation and development of the shoot apical meristem have been performed on meristems that developed during embryogenesis. We characterized the in situ formation of adventitious shoots originating from cells located near leaf axils in a series of six interspecific periclinal tobacco chimeras. Shoots were generated by decapitating the plants and removing all of the axillary buds. Eighty-four of the 413 shoots regenerated were chimeral. Many of the shoots were complex mericlinal chimeras, with several of their axillary buds possessing meristems arranged as periclinal chimeras. The adventitious shoots originated from derivatives of only the second and third cell layers in the meristem of the source plant since derivatives of the outermost meristem layer were removed with the removal of the axillary buds. With time, nearly all adventitious shoot apices that were initially chimeral became nonchimeral or stabilized as periclinal chimeras. A statistical analysis indicated that the arrangement of genetically distinct tissues in the cell layers of adventitious meristems was influenced by competition between cell types. Our method for generating these shoots can be used to create small genetically distinct sectors analogous to radiation-induced sectors as well as a complete series of periclinical chimeras, both of which have potential for use in determining tissue to-tissue interactions. An analysis of sector length and position on chimeral shoots indicates that the first one to three leaves of adventitious shoots do not arise as derivatives of the shoot apical initials of a meristem proper. PMID- 8416839 TI - Development of the taste bristles on the labellum of Drosophila melanogaster. AB - The taste bristles of the adult labellum develop from the labial discs within the first 30 hr of pupation. The neuron-specific antibody Mab22C10 and the tissue specific beta-galactosidase activity in the A37 strain were used as cell markers for the developing sensilla. These experiments revealed that the sensory progenitors of all the labellar bristles are specified in three waves occurring 0, 6, and 16 hr after pupation. The incorporation of the thymidine analog 5-bromo 2'-deoxyuridine (BrdU) into the cells of the developing proboscis further supports the temporal pattern of the sensillum specification. Mitotic activity in the bristles specified in the first wave is completed before the next wave is initiated. The eight cells that constitute a single bristle share a common lineage. The trichogen and tormogen cells are siblings from a single progenitor and the thecogen and the five neurons share a common ancestry. Our results provide a basis for the identification of molecules that specify the development of the polyinnervated bristles of the adult and the specification of neurons with different stimulus specificities. PMID- 8416840 TI - Early amphibian (anuran) morphogenesis is sensitive to novel gravitational fields. AB - Anuran amphibian embryos (Xenopus laevis and Rana dybowskii) are sensitive to novel gravitational fields. Under simulated weightlessness, (i) the location of the first horizontal cleavage furrow was shifted toward the vegetal pole at the eight-cell stage; (ii) the position of the blastocoel was more centered, and the number of cell layers in the blastocoel roof was increased at the blastula stage; (iii) the dorsal lip appeared closer to the vegetal pole at the gastrula stage; and (iv) head and eye dimensions were enlarged at the hatching tadpole stage. Effects of simulated hypergravity were opposite to those of simulated weightlessness, except that hypergravity, unlike simulated weightlessness, reduced the number of primordial germ cells in feeding tadpoles. Despite those dramatic differences in the early embryogenesis, tadpoles at the feeding stage are largely indistinguishable from controls. PMID- 8416841 TI - 43K protein and acetylcholine receptors colocalize during the initial stages of neuromuscular synapse formation in vivo. AB - The 43K protein is a cytoplasmic peripheral membrane protein concentrated subsynaptically in skeletal muscle. Recombinant 43K has been shown to cause clustering of acetylcholine receptors (AChRs) in cultured cells. However, the role of 43K in vivo is disputed, because in some cases it appears only after AChRs have clustered. We therefore examined the expression and distribution of 43K and AChRs during synapse formation in embryonic mouse muscles. Messenger RNA for 43K was detected on Embryonic Day (E) 12, a day prior to the first AChR clusters. Immunofluorescence showed that both AChRs and 43K were colocalized in patches by E13, the stage at which intramuscular nerves were first detected. The AChR/43K patches were nerve associated, and more than 98% of AChR patches were accompanied by 43K. The precise colocalization of 43K and AChRs persisted through development. These results are consistent with 43K being involved in the nerve induced clustering of AChRs during synapse formation. PMID- 8416842 TI - Cranial neural crest cells synthesize and secrete a latent form of transforming growth factor beta that can be activated by neural crest cell proteolysis. AB - Transforming growth factor beta (TGF-beta) is an important regulator of cell growth and differentiation. TGF-beta is usually secreted in a latent form (i.e., not biologically active) that can be activated by limited exposure to low pH or specific proteolytic cleavage. In this study, we (1) assayed cranial neural crest (NC) cell-conditioned medium for the presence of active and latent TGF-beta, (2) determined whether TGF-beta was activated by NC-generated plasmin, and (3) examined whether active TGF-beta 1 regulates NC cell plasminogen activator activity. Results show that under serum-free conditions, essentially all of the TGF-beta secreted by NC cells is in a latent form. However, 24 hr after adding plasminogen to the cultures, active TGF-beta was detectable. Treatment of NC cells with active TGF-beta 1 significantly decreased NC cell plasminogen activator activity. These data suggest that NC cells secrete a latent form of TGF beta that can be activated under conditions favoring the generation of local proteolytic activity and that levels of plasminogen activator activity may be autoregulated via an autocrine effect of this growth factor. PMID- 8416843 TI - 3T3 cell integration and differentiative potential during limb development in the mouse. AB - In this study we have investigated the ability of embryonic mouse limb cells to induce the differentiation of 3T3 cells both in vivo and in vitro. Using the exo utero surgical technique we introduced labeled 3T3 cells into the embryonic mouse limb and monitored their subsequent development. We found no indication that the normal pattern of tissue differentiation was perturbed; rather we observed that 3T3 cells were closely associated with a variety of limb mesodermal tissues which include muscle, tendon, and ligament. In very rare instances did we observe 3T3 cells associated with cartilage. To further investigate the differentiative potential of 3T3 cells we cocultured labeled 3T3 cells with limb cells under conditions that favored either cartilage or muscle differentiation. The results from these experiments suggest that 3T3 cells avoid regions of chondrogenesis and rarely participate in myotube formation. We conclude that 3T3 cells are not responding to differentiation signals present in the developing limb, but rather are responding to signals involved in growth and tissue morphogenesis. PMID- 8416844 TI - Overlapping expression of Xwnt-3A and Xwnt-1 in neural tissue of Xenopus laevis embryos. AB - Xwnt-3A is a member of the Xenopus-Wnt gene family, a class of secreted, cysteine rich proteins implicated in intercellular signaling during early development. Here we describe the full-length coding sequence of Xwnt-3A, as well as the spatial expression pattern of this Xwnt gene as determined by whole-mount in situ hybridization analysis. While Xwnt-3A shares considerable amino acid identity with both Wnt-3 (87%) and Wnt-3A (85%), its spatial expression pattern is most like that of Wnt-3A. Xwnt-3A, which is first detected at the neurula stage of development, is expressed exclusively along the dorsal midline of the developing brain and neural tube and along the dorsal surface of the otic vesicle. While the expression of Xwnt-1 extensively overlaps that of Xwnt-3A, Xwnt-1 is uniquely expressed along the midbrain/hindbrain boundary and is absent from the otic vesicle. The expression of Xwnt-3A in neural ectoderm is dependent upon neural induction as determined by experiments with recombined ectoderm and mesoderm tissue. These results suggest that Xwnt-3A may participate in patterning the central nervous system during early Xenopus development. Last, the ectopic expression of Xwnt-3A induces the formation of a secondary axis at the anterior end of the embryo. PMID- 8416845 TI - Centrosome inheritance in starfish zygotes. II: Selective suppression of the maternal centrosome during meiosis. AB - Although both gametes may contribute a centrosome to the zygote at fertilization, only one of these centrosomes is used in development. Thus, specific mechanisms must exist to control centrosome inheritance in all sexually reproducing organisms. We use starfish as a model system to characterize these control mechanisms because the eggs complete meiosis I and meiosis II after fertilization; this allows us to directly follow the fate of all parental centrosomes in vivo. Only the paternal centrosome is used in starfish development. Although the microtubule organizing center activity of the maternal centrosome persists, the functional loss of this centrosome involves the suppression of its ability to double, or reproduce, at successive mitoses (Sluder et al., 1989. Dev. Biol. 131, 567-579). To determine when the reproductive capacity of the maternal centrosome is degraded, we transfer meiosis I and meiosis II spindles from just fertilized eggs into other zygotes that are in prophase of first mitosis. Meiosis I spindles are stable during first mitosis and are disassembled in first telophase in concert with the host spindle. In 61% of the cases a variable number of formerly meiotic centrosomes are active at second mitosis and reproduce in a normal fashion between subsequent mitoses. However, when meiosis II spindles are transferred in the same manner, in only 26% of the cases do any of the centrosomes persist past first mitosis or reproduce in a normal fashion thereafter. In the remainder of the cases the remnants of the maternal centrosomes organize a single monaster that does not double between mitoses. Control transfers of first mitosis spindles indicate that these results are not due to nonspecific damage to the meiotic spindles or to the recipient zygotes. These observations indicate that the reproductive capacity of maternal centrosomes is degraded during meiosis I, not during oogenesis. Our results also show that the cytoplasmic conditions which eliminate this reproductive capacity are no longer active once the zygote has entered the first mitotic cell cycle. PMID- 8416846 TI - Direct induction of DNA hypermethylation in sea urchin embryos by microinjection of 5-methyl dCTP stimulates early histone gene expression and leads to developmental arrest. AB - The role of DNA methylation in gene inactivation has been studied extensively in vertebrates but it is not clear whether it serves similar functions in other organisms. We devised a novel approach to induce hypermethylation of both endogenous and injected DNA in the sea urchin Lytechinus pictus in order to study the effect of DNA methylation on gene expression in this invertebrate. By injecting 5-methyl dCTP either alone or together with a cloned DNA construct into fertilized sea urchin eggs, replicating DNA became hypermethylated from the random incorporation of the methylated nucleotide in place of cytosine during DNA synthesis. During subsequent rounds of replication, the injected 5-methyl dCTP became depleted but methylation at CpG sites was still elevated presumably due to the action of a methyltransferase enzyme. Using this approach, we studied the effect of hypermethylation on two members of the sea urchin multigene family, the early H2B and the late H2B genes. De novo methylation was shown to occur at known cis-regulatory regions of the genes. The effect of methylation on gene activity was probed using RNase protection assay. Methylation resulted in increased early H2B histone gene expression but had no effect on late H2B histone gene expression. These results demonstrate that methylation does not necessarily inactivate genes in the sea urchins as previously thought. Interestingly, the development of embryos injected with 5-methyl dCTP typically was arrested at the blastula stage, and analysis of the genomic DNA extracted from injected embryos showed a significant increase in the endogenous methylation content. These data suggest that perturbation of methylation patterns in developmental sea urchin embryos may be responsible for the developmental arrest through altering the gene expression pattern. PMID- 8416847 TI - Localization and quantification of Wnt-2 gene expression in mouse mammary development. AB - The Wnt gene family encodes a group of proteins probably involved in cell-cell communication during several stages of vertebrate development. More than 10 members of this family have been identified and shown to be expressed mainly in developing neural tissue. Using a reverse transcription-polymerase chain reaction (RT-PCR)-based approach with degenerate oligonucleotides directed against conserved sequences in the Wnt genes, Wnt-2 transcripts were detected in RNA isolated from mammary glands of 4- to 6-week-old virgin C3H mice, a period characterized by extensive end bud and ductal proliferation. The spatial and temporal expression of Wnt-2 in the developing mouse mammary gland was studied by in situ hybridization, quantitative RT-PCR, and Northern analysis. Wnt-2 is expressed during the ductal phase of mammary gland development, primarily in the basal layer of mammary ducts and in the body cells of end buds. Wnt-2 RNA transcripts were readily detected in poly(A) RNA isolated from 5-week-old C3H and Balb/c mice. RNA transcript levels measured as molecules per nanogram of total RNA by RT-PCR decreased 10- to 40-fold within 2 days after the onset of pregnancy and remained low during pregnancy and lactation. This is in contrast to the patterns of expression of other Wnt family members, Wnt-5a and -5b, whose expression was either barely or not detectable in the 4- to 6-week-old mammary gland, but increased markedly during pregnancy. These results confirm the differential expression of Wnt gene family members during mammary gland development. Furthermore, they suggest that Wnt-2, as well as several other family members, may play a role in pattern formation during early mammary gland development. PMID- 8416848 TI - Heparan sulfate proteoglycan (perlecan) expression by mouse embryos during acquisition of attachment competence. AB - Cell surface expression of the basement membrane form of heparan sulfate proteoglycan (HSPG), perlecan, basic fibroblast growth factor (bFGF) binding forms of heparan sulfate, and laminin has been examined by immunofluorescence during the attachment and outgrowth phases of mouse embryo development in vitro and in utero. The results indicate that both HSPG and laminin are expressed at the exterior surface of the trophectoderm during the attachment phase of implantation in vivo. HSPG expression correlated with acquisition of attachment competence by embryos in vitro. The intensity of immunofluorescence signal appeared to decrease by the outgrowth stage of development in vitro. It is concluded that expression of HSPG at the trophectodermal surface is coordinated with development of attachment competence by mouse embryos in vitro and in utero. PMID- 8416849 TI - DentaScan imaging of the mandible and maxilla. AB - DentaScan is a unique new computer software program which provides computed tomographic (CT) imaging of the mandible and maxilla in three planes of reference: axial, panoramic, and oblique sagittal (or cross-sectional). The clarity and identical scale between the various views permits uniformity of measurements and cross-referencing of anatomic structures through all three planes. Unlike previous imaging techniques, the oblique sagittal view permits the evaluation of distinct buccal and lingual cortical bone margins, as well as clear visualization of internal structures, such as the incisive and inferior alveolar canals. Several case reports are presented to demonstrate the clinical usefulness of DentaScan. PMID- 8416850 TI - Management of squamous cell carcinoma of the floor of mouth. AB - Between 1964 and 1987, 194 patients with previously untreated squamous cell carcinoma of the floor of mouth were managed at the University of Florida. A retrospective analysis was undertaken in order to evaluate the treatment results and associated complication rates. Surgery or irradiation alone was found to result in similar local control rates for stage I and II lesions, whereas more advanced tumors had better local control rates with a combination of surgery and irradiation. Radiotherapy had a higher incidence of minor and moderate complications, whereas a greater number of severe complications occurred after surgery. We recommend surgery for early lesions due to the lower overall incidence of associated complications. Despite a higher risk of severe complications, combination therapy is recommended for more advanced lesions due to improved local control as compared to single modality therapy. PMID- 8416851 TI - Association of hyperparathyroidism with nonmedullary thyroid carcinoma: review of 31 cases. AB - In a series of 948 patients operated on for primary hyperparathyroidism (HPT) by one surgeon (JNA) from 1952 to 1992, there were 242 (26%) instances of coincidental thyroid and parathyroid disease. Of these, 211 had benign thyroid lesions. In the remaining 31 cases HPT was associated with nonmedullary thyroid carcinoma; all were treated by resection of parathyroid adenomas and thyroidectomy. One patient died of unrelated cause (carcinoma of breast) 11 years following surgery. The remaining 30 patients are living and well 2 to 20 (mean 8.2 years) years after surgery; there was no recurrence or thyroid cancer-related mortality in the series. In contrast to prior reports, only six (20%) of our patients had a history of prior radiotherapy. We suggest that during neck exploration for HPT, the entire thyroid gland be evaluated and all palpable nodules resected and submitted to pathologic study. PMID- 8416852 TI - Surgery for parathyroid adenoma and hyperplasia: relationship of histology to outcome. AB - Recent histopathologic evidence challenges the teaching that enlargement of a solitary parathyroid gland is invariably caused by an adenoma, whereas multiple gland enlargement results from hyperplasia. We have re-examined the parathyroid tissue obtained from 152 consecutive patients undergoing surgery for primary hyperparathyroidism and compared it with their clinical outcome. Our approach was to excise enlarged glands and biopsy the remainder. In 124 patients (82%) at least three glands were biopsied or removed. The ratio of adenoma to hyperplasia was reversed by our histologic re-examination; adenomas were found in only 27 patients (25 single, two double), whereas hyperplasia was found in 117 patients (one gland, 87 patients; two glands, 16 patients; three glands, five patients; four glands, nine patients). Normal tissue only was reported in eight patients. During a 2-year follow-up, five patients (3%) developed hypocalcemia and none developed recurrent hypercalcemia. Our results indicate that a full neck exploration with removal of all enlarged glands is more important than the histologic diagnosis in planning a successful surgical strategy for primary hyperparathyroidism. PMID- 8416853 TI - Local control of squamous cell carcinoma following marginal and segmental mandibulectomy. AB - The records of 130 consecutive patients undergoing marginal or segmental mandibulectomy for squamous cell carcinoma of the oral cavity or oropharynx were reviewed. An attempt was made to correlate incidence of recurrence with characteristics of the primary tumor and extent of mandibulectomy. The local recurrence rate was 19% following marginal mandibulectomy and 6% following segmental mandibulectomy. The incidence of local recurrence was independent of the size of the primary tumor or the extent of lymph node metastases. Neither mandibular invasion by tumor nor the addition of radiotherapy influenced local recurrence. Ten of 15 patients recurring locally after marginal mandibulectomy were salvaged by further surgery. This study suggests that local control following marginal mandibulectomy is comparable to that following segmental mandibulectomy regardless of the size of the tumor. Segmental mandibulectomy should be reserved for those tumors invading deeply into the mandible or wrapping around it. PMID- 8416854 TI - Accuracy of intraoperative frozen section diagnosis in head and neck surgery: experience at a university medical center. AB - We performed 2,210 intraoperative frozen sections on 258 patients from the Otolaryngology-Head and Neck Surgery Service in 1990 and 1991. Surgery involved a wide variety of benign and malignant lesions. Techniques included biopsies for diagnosis, simple excision, thyroid and salivary gland surgeries, lymph node biopsies, composite resections with radical neck dissections, laryngectomies, and skull base surgeries. During the 2-year period, 1,947 (88.1%) sections were requested for evaluation of surgical margins, 258 (11.7%) for diagnosis, and five (0.2%) cases for tissue identification. There was disagreement between the frozen section and permanent section in 46 (2.1%) cases, and the deferral diagnosis rate was 0.8%. Disagreements were the result of sampling errors in 33 and interpretive errors in 13 cases. There were six (0.3%) false-negative diagnosis of malignancy and four (0.2%) false-positive diagnoses of malignancy. Three of these had an impact on patient care as discussed. We recommend careful sampling and sectioning of small biopsies and the need for vigilant communication between surgeon and pathologist. PMID- 8416855 TI - Primary carcinoma involving the petrous temporal bone. AB - Between 1975 and 1985, 29 patients with the diagnosis of carcinoma of the petrous temporal bone were seen at the Princess Margaret Hospital. Twenty-seven carcinomas were graded: 13 were well-differentiated; the remaining 14 were either moderately or poorly differentiated tumors. Fifteen patients were managed with a combination of surgery and radiotherapy, 13 were treated with radiotherapy only, and one patient was treated by surgery alone. Median follow-up time was 6.1 years, and the 5-year actuarial local control and cause-specific survival rates for the entire group were 40% and 50%, respectively. Age greater than 60 years, poor grade of tumor, and involvement of the facial nerve were three significant variables associated with poor outcome. A superior 5-year actuarial local control was achieved with surgery plus postoperative radiotherapy (54%) compared to other treatment approaches. Based on the results from this review, we would continue to recommend a combined modality approach of surgery followed by postoperative radiotherapy in the management of this rare, but life-threatening disease. PMID- 8416856 TI - Relative roles of computed tomography and endoscopy for determining the inferior extent of pyriform sinus carcinoma: correlative histopathologic study. AB - Ten laryngopharyngectomy specimens were dissected to determine the relative accuracy of computed tomography (CT) and endoscopy under anesthesia for evaluating the inferior extent of pyriform sinus carcinoma. Endoscopic examination failed to detect involvement of the pyriform sinus apex in one patient, and considerably underestimated disease at this level in two others; CT accurately predicted the status of the apex in these three and all other patients. Endoscopy underestimated the inferior extent of tumor in six patients; CT revealed the inferior limit relative to the esophageal verge more accurately in all six of these submucosal extensions. Endoscopy revealed one case of mucosal spread to the esophageal verge not demonstrated by CT. Endoscopy and CT were both correct in revealing the esophageal verge to be involved (one case) and free of disease (two cases). This study also confirmed a common tendency of pyriform fossa cancer to spread through thyrohyoid membrane adjacent to the course of superior laryngeal neurovascular bundle (six cases). High-resolution CT, in experienced hands, is a useful adjunct to endoscopy for detecting submucosal, inferior extension of pyriform sinus carcinoma. This information can influence the choice of the lower margin of resection and method of pharyngeal reconstruction. PMID- 8416857 TI - Actinomycosis of the tongue: a diagnostic challenge. AB - Actinomycotic infections of the cervicofacial area are rare and frequently show a confusing clinical picture. We report a case of actinomycosis that mimicked a neoplasm of the tongue. The clinical and pathologic features and the differential diagnosis of this unusual oral infection are discussed. PMID- 8416858 TI - Cervical and mediastinal abscess as a manifestation of histoplasmosis. AB - Histoplasmosis is the most common endemic respiratory mycosis in the United States. Disseminated histoplasmosis generally is initially seen with systemic or respiratory complaints. Presentation of histoplasmosis as a neck mass is rare. Due to this unusual presentation and the diagnostic implications, we present a case of disseminated histoplasmosis that presented as a neck abscess. Unusual presentations of histoplasmosis may occur even in patients who are not immunocompromised. The diagnosis and management of histoplasmosis is reviewed. Histoplasmosis should be considered in the workup of a cervical abscess even in the immunocompetent host. PMID- 8416859 TI - Phaeohyphomycosis of the paranasal sinuses masquerading as a neoplasm: a case report. AB - A case of fungal infection of the paranasal sinuses simulating a neoplasm is reported. Fonsecaea pedrosoi, a dematiacious fungus, which has not been previously reported, was responsible for this unusual infection. Fungal infection may mimic malignant lesions of the paranasal sinuses; however, coexisting malignancy must be ruled out. PMID- 8416860 TI - Cavernous sinus involvement in head and neck cancer. AB - Spread of tumor to intracranial structures is an infrequent and late manifestation of head and neck cancers. We recently encountered six patients with a distinct clinical syndrome due to involvement of the cavernous sinus, which forms the basis of this report. This syndrome was a source of significant morbidity and mortality, with a mean survival of only 4 months. The diagnosis is often elusive, but is now made more commonly than previously. Whether this reflects increased incidence (due to alterations in the natural history of disease by therapy) or improved diagnosis (due to modern imaging modalities) is unknown. Cavernous sinus involvement may be the first evidence of distant disease in head and neck cancer. Although survival is poor, palliation is worthwhile. Awareness of this syndrome can lead to earlier diagnosis and alteration of treatment. PMID- 8416861 TI - Acinic cell carcinoma of the parapharyngeal space: case report. AB - We describe a case of acinic cell carcinoma (ACC) of the parapharyngeal space in a 58-year-old woman. A tumor originating from the deep lobe of the parotid gland was totally excised by an external cervical approach. The occurrence of ACC in the parapharyngeal space is extremely rare. We discuss the management, especially surgical procedure, of ACC in this area. PMID- 8416862 TI - Metastatic head and neck carcinoma in a percutaneous gastrostomy site. AB - Two unusual cases of metastatic head and neck carcinoma to the exit site of a percutaneous endoscopic gastrostomy (PEG) tube are reported. These patients presented with squamous cell carcinomas of the supraglottic larynx and oropharynx. In both patients, a PEG tube was inserted using the "pull" technique prior to any extirpative measures. Within 9 months after surgical treatment, each patient was diagnosed with metastatic squamous cell carcinoma at the PEG tube exit site. A review of the literature revealed three cases of metastatic head and neck neoplasms to a PEG site and a single case report of spread to the wound of an "open" gastrostomy. The hematogenous route is the probable mechanism of metastasis, but direct implantation cannot be dismissed. Until further data become available, the authors recommend that, when possible, PEG tube placement using the "pull" technique be deferred until after extirpation of head and neck tumors. PMID- 8416863 TI - Immunology of head and neck cancer: perspectives. PMID- 8416864 TI - Neural damage after carotid artery surgery. PMID- 8416865 TI - Evaluation of microwave interstitial antennas in the phantom with varying cross section. AB - Dipole-regular microwave interstitial antennas are characterized with a "dead" space located along the tip segment of the antenna. The length of the "dead" space is on the order of 2 cm or larger, depending on the antenna's insertion depth. If the insertion depth is smaller than 4 cm, then coupling of the antennas to tissue becomes a problem. Catheters that facilitate the placement of antennas into tumor frequently protrude beyond the tissue. This provides the opportunity of exposing part of the antenna tips (with low radiation output) beyond the tissue. Decoupling of this part of the antennas from the tissue reduced the dead space and improved microwave power transfer to the tissue. This concept was investigated using a muscle equivalent phantom consisting of five segments with thicknesses varying from 3 cm to 8 cm. The transfer of microwave power to the phantom and SAR distributions along the central axis of a rectangular array of four antennas were evaluated by measuring rates of temperature rise. The protrusion lengths that improved the array performance were found for each segment of the phantom. PMID- 8416866 TI - Ethmoidal cancers: a retrospective study of 22 cases. AB - From April 1978 to June 1990, 22 patients with ethmoidal cancer were treated at Fondation Bergonie by a combination of surgery and radiation therapy. The mean age was 59.6 years (range 34-79 years) and the sex ratio is 2.7 (16 males/6 females). Histologic types were: adenocarcinoma, 13 cases; squamous carcinoma, 4 cases; undifferentiated carcinoma, 3 cases and esthesioneuroblastoma, 2 cases. Exposure to wood dust was encountered in 11 patients, especially in cases of adenocarcinoma: 10/13 (77%). Staging according to the classification of the University of Florida was: Stage I, 10 patients; Stage II, 5 patients and Stage III, 7 patients. Resection was considered as complete in 16 cases and only one orbital exenteration was performed. The postoperative radiation therapy delivered a mean given dose of 55.7 Gy (range 50-70 Gy) expressed to the hot spot using a technique adapted to tumor location and extension. Complete remission was achieved in 20 cases. Median follow-up is 28 months. The 5-year overall and disease-free survival are 44% and 38%, respectively. Analysis of recurrences according to staging gives: 5/10 Stage I, 2/5 Stage II and 5/7 Stage III. Recurrence is pejorative since death occurs in all cases within an average of 6 months following salvage treatment, except for three patients still alive within less than 6 months and in second remission. Prognosis of ethmoidal cancer depends on staging and local control. PMID- 8416867 TI - Measurement of mechanical accuracy of isocenter in conventional linear accelerator-based radiosurgery. AB - PURPOSE: Five Varian linear accelerators were studied to determine whether their mechanical isocentric accuracies were sufficient for radiosurgery and, if not, if the observed errors were sufficiently consistent and predictable to be correctable by some form of secondary collimator steering device to maintain isocentric alignment. METHODS AND MATERIALS: A 0.3 mW 670 nm diode laser was mounted in the secondary collimator insert of a radiosurgery extended collimator assembly. A cylindrical lens was used to create a laser fan beam that passed through isocenter and could be oriented parallel or perpendicular to the plane of rotation. A position sensitive photo-diode having an electrical output that varied with the portion of its surface illuminated was mounted at isocenter in a rotational mount. This mount tracked the accelerator gantry such that the surface of the photo-diode remained perpendicular to the laser beam during gantry rotation. An X/Y recorder was connected to the gantry-angle potentiometer of the accelerator and to the photo-diode and plotted the positional variation from isocenter with gantry rotation. RESULTS: The root-mean-square error for the five machines was +/- 0.06 to +/- 0.08 mm in the plane of rotation and +/- 0.17 to +/- 0.35 mm out of (perpendicular to) the plane of rotation. The in-plane-of-rotation errors tended to be maximal near the diagonal gantry angles and the out-of-plane of-rotation errors were maximal in the over and under vertical positions. CONCLUSIONS: Both types of errors were predictable but only the out-of-plane-of rotation errors were considered large enough to warrant consideration of correction (although the need is debatable). On all the tested machines, the out of-plane-of-rotation error curve was a relatively smooth bell-shaped function that would be readily amenable to correction. The diode laser/photo-detector system used should prove useful in accurately defining isocenter and facilitating the precise adjustment of the laser isocenter lights. PMID- 8416868 TI - VISTAnet: interactive real-time calculation and display of 3-dimensional radiation dose: an application of gigabit networking. AB - Three-dimensional treatment planning can allow the clinician to create plans that are highly individualized for each patient. However, in lifting the constraints traditionally imposed by 2-dimensional planning, the clinician is faced with the need to compare a much larger number of plans. Although methods to automate that process are being developed, it is not yet clear how well they will perform. VISTAnet is a 3 year collaborative effort between the Departments of Radiation Oncology and Computer Science at the University of North Carolina, the North Carolina Supercomputing Center, BellSouth, and GTE with the medical goal of providing real-time 3-dimensional radiation dose calculation and display. With VISTAnet technology and resources, the user can inspect 3-dimensional treatment plans in real-time along with the associated dose volume histograms and can fine tune these plans in real-time with regard to beam position, weighting, wedging, and shape. Thus VISTAnet provides an alternate and, possibly, complementary approach to computerized searches for optimal radiation treatment plans. Building this system has required the development of very fast radiation dose code, methods for simultaneously manipulating and modifying multiple radiation beams, and new visualizations of 3-dimensional dose distributions. PMID- 8416869 TI - Anatomy of the celiac axis and superior mesenteric artery and its significance in radiation therapy. AB - PURPOSE: In the radiation therapy of upper gastrointestinal malignancies, treatment of lymph nodes in the region of the celiac axis and superior mesenteric axis is often mandated. This study was undertaken to determine the relationship of the celiac axis and superior mesenteric arteries to the vertebral bodies--the radiographically visualized reference structures during simulation. METHODS AND MATERIALS: Twenty-three celiac angiograms and 24 superior mesenteric angiograms performed preoperatively in 24 patients treated at the University of Pennsylvania from 1984 to 1989 for pancreatic carcinoma were examined. The location of the origin of the celiac and superior mesenteric arteries was determined in each case. RESULTS: In 48% of the celiac angiograms, the celiac axis arose from the aorta high at the pedicle of the T-12 vertebral body, contrary to the common belief that the celiac axis arises near the T12-L1 interspace. The superior mesenteric artery arose at the level of L-1 in 83% of the 24 angiograms and below the pedicle of L-1 in 5 (21%). However, none arose below the L1-2 interspace. CONCLUSION: The variability demonstrated in the levels from which these vessels arise strongly suggests individualized treatment planning, including angiographic, CT or MRI data should be performed if tight margins are used. These studies would additionally optimize treatment of the tumor bed. Consideration for dose at field edges (i.e., "buildup") and day to day variation in set-up is required in determining the field borders. Treatment volumes tightly encompassing T12 and L1 could risk undertreating regional lymph nodes associated with these vessels. PMID- 8416870 TI - Modeling of normal tissue response to radiation: the critical volume model. AB - PURPOSE: A model for calculating normal tissue complication probability in response to therapeutic doses of radiation is presented. METHODS AND MATERIALS: The model which we call the "critical volume model" is based on a concept of functional subunits defined either structurally (e.g., nephrons) or functionally, and an assumption that normal tissue complication probability is fully determined by the number or fraction of surviving functional subunits composing an organ or tissue. The essential features of the model are that it takes into account variations in tissue radiosensitivity and architecture of an organ for a single patient and for a patient population, and predicts the normal tissue complication probability under conditions of 3-dimensional inhomogeneity of the dose distribution. The model can be used for Integral Response, or "parallel," organs (where all functional subunits are performing the same function in parallel and the output of the organ is the sum of the outputs of the functional subunits and for Critical Element, or "serial," organs (where damage to one functional subunit results in an expression of damage for the whole organ). The model combines into one compact scheme new concepts and several ideas and models which have been previously developed by other investigators. RESULTS: The behavior of the model is presented and discussed for the example of the kidney, with clinical nephritis as the functional endpoint. CONCLUSIONS: The model has the potential to be a useful tool for evaluation and optimization of 3-dimensional treatment plans for a variety of types of normal tissues. PMID- 8416871 TI - Cellular radiosensitivity as predictors of treatment outcome: where do we stand? PMID- 8416872 TI - Relative clinical influence of tumor dose versus dose per fraction on the local control and late normal-tissue morbidity after larynx radiotherapy. PMID- 8416873 TI - Critical volume model. PMID- 8416874 TI - Radiation as adjunctive therapy to cystectomy for bladder cancer. PMID- 8416875 TI - Should tumors be clamped in radiobiological fractionation experiments? PMID- 8416876 TI - Postoperative radiation therapy for squamous cell carcinomas of the oral cavity and oropharynx: impact of therapy on patients with positive surgical margins. AB - PURPOSE: The presence of a positive or close margin after resection of a squamous cancer of the head and neck is associated with a significant risk of local recurrence. To determine the efficacy of postoperative radiation therapy for patients with advanced oral cavity and oropharyngeal cancers with inadequate margins of resection, the present retrospective analysis was undertaken. METHODS AND MATERIALS: One hundred and two patients were treated with surgery and postoperative radiation therapy for advanced squamous cell carcinomas of the oral cavity and oropharynx. The anatomic subsites treated include oral tongue (n = 29), floor of mouth (n = 22), base of tongue (n = 31) and tonsillar fossa (n = 20). Twenty-five patients (25%) had positive margins, 41 patients (40%) had close margins (< or = 0.5 cm from the surgical margin) and 36 (35%) had negative margins. The median radiation dose was 6000 cGy. RESULTS: With a median follow-up of 7 years, the actuarial control rate for patients with positive, close and negative margins was 79%, 71%, and 79%, respectively. When postoperative doses of > or = 60 Gy were delivered to patients with positive/close margins (excluding patients with oral tongue lesions), the 7-year actuarial control was 92%. In similar patients receiving < 60 Gy, the actuarial control was 44% (p = 0.0007). Compared to other anatomic subsites, inferior control rates were obtained with oral tongue lesions. For this subsite, the control rates for positive, close, and negative margins were 50%, 62% and 69% respectively. CONCLUSION: We conclude that excellent local control can be achieved with postoperative radiation therapy, despite the presence of inadequate margins of resection, when doses of > or = 60 Gy are used. Future strategies must be directed at further improving these results in patients with oral tongue lesions. PMID- 8416877 TI - Relative clinical influence of tumor dose versus dose per fraction on the occurrence of late normal tissue morbidity following larynx radiotherapy. AB - A study was made of 242 cases of T2 No glottic cancer treated by definitive radiotherapy over a 15-year period. The aim was to examine the relative influences of tumor biological dose (indicated by locoregional control) versus dose per fraction on the occurrence of late normal tissue effects; in addition, the impact of cord mobility on outcome was analyzed. The 5-year survival corrected for intercurrent deaths was 84% and local disease-free (larynx and/or nodes) survival was 76%. Using Cox regression analysis the only factor significant for local control was cord mobility (p < 0.0001) which also had an effect on overall survival (p < 0.0001); subdivision of T2 glottic staging should be reintroduced into staging classifications. It is evident that comparison of clinical results between centers is potentially prejudiced by an array of factors relating not only to fractionation differences but also variation in clinical and organizational aspects of care. Nevertheless, using other published data for comparison, it seems likely that the serious morbidity rate of 4.1% seen in this study is due in some part to the high tumor biological dose (resulting in high local control). The influence of fraction size was difficult to discern as equivalence in local control was not seen in the data chosen. PMID- 8416878 TI - Results of cesium needle interstitial implantation for carcinoma of the oral tongue. AB - One hundred thirty previously untreated patients with invasive squamous cell carcinoma of the oral tongue received interstitial radiotherapy with curative intent using cesium needles. Ninety-nine patients were treated with interstitial radiotherapy alone and 31 patients received interstitial radiotherapy combined with external beam irradiation. The recurrence-free rates in the primary lesions were 94.4% (17/18) in T1, 91.2% (52/57) in T2, and 70.9% (22/31) in T3 lesions. The local recurrence-free rates with single-plane and two-plane implantation were good: 89.7% (70/78) and 85.7% (12/14), respectively. The rate of 64.2% (9/14) for volume implantation was significantly poorer (p < 0.05). It is evident that tumor volume is an important factor in the control of cancer following interstitial therapy. The overall incidence of ulceration of the tongue and mandibular complication was 20% (26/130) and 13% (17/130), respectively. Using both interstitial and external radiotherapy, the incidence was 22.5%, compared with 10.1% using interstitial radiotherapy alone. The mandibular complication incidence of 8.9% with single-plane implants was much lower than 20.8% for two plane and 23.5% for volume implants. Interstitial radiotherapy is most suitable in T1 and T2 cases in which single-plane implantation is possible; for these patients interstitial radiotherapy, which has the advantage of preserving the structure and function of the tongue, should continue to be used in the future in spite of the progress in reconstructive surgery. PMID- 8416879 TI - Predictive value of in vitro radiosensitivity parameters in head and neck cancers and cervical carcinomas: preliminary correlations with local control and overall survival. AB - PURPOSE: To determine whether in vitro radiosensitivity parameters are predictive of treatment outcome. METHODS AND MATERIALS: Biopsies were obtained from patients with head and neck cancers (57) and cervical carcinomas (20) and in vitro radiosensitivity parameters were obtained using the CAM plate assay. RESULTS: In most cases (75%) patients were treated with radiation alone. The median follow up was 461 days. When the whole group of head and neck cancers and cervical carcinomas was considered, patients with a SF2 value below 0.36 had a higher 2 year local control rate (93% versus 68%) and a higher 2-year survival rate (71% vs. 62%) than those with SF2 values above that threshold, but differences were not significant. These trends persisted when head and neck cancers were considered alone with a higher local control rate (86% vs. 67%) and a higher survival rate (75% vs. 52.5%) obtained for patients with a SF2 value below 0.36. When the alpha value was evaluated for the whole group of patients a significantly higher local control rate (80.5% vs. 40.5%) and overall survival rate (71% versus 37.5%) at 2 years were obtained for patients with alpha values above 0.07 Gy-1. When only the group of head and neck cancers was considered, local control rate was significantly higher (79% vs. 33%) but overall survival rate (65.5% vs. 33%) was not significantly higher for alpha values above 0.07 Gy 1. CONCLUSION: These results are encouraging but need to be confirmed with a larger number of patients with a longer follow-up. PMID- 8416880 TI - The efficacy of radiation therapy for a malignant melanoma in the mucosa of the upper jaw: an analytic study. AB - An analysis has been made of the effect of radiation therapy in 28 patients with a malignant melanoma (Stage I: 18 cases; Stage II: 10 cases) in the mucosa of the upper jaw. Treatment had been provided by one of the following methods: intraoral mold (10 cases), interstitial brachytherapy (two cases), intraoral electron therapy (nine cases), or external irradiation (seven cases). The results have shown that the survival rate for all 28 patients was 25%, and that the survival rate for stage I patients treated by intraoral electron or brachytherapy was 47%. The primary tumor control rate in percentages was 79% (22/28) in all 28 radiotherapy patients; 92% (11/12) for tumors treated by a mold or an interstitial implant; 67% (6/9) for tumors treated by an intraoral cone; and 71% (5/7) for tumors treated by external irradiation with or without surgery. A neck metastasis that was found in 19 patients was treated by surgery, radiotherapy, and/or immunochemotherapy, and the result was successful in nine patients. The major factor in the failure of treatment was a distant, metastatic dissemination. This analysis revealed that radiotherapy has achieved similar or better results than surgery and may be advocated for the management of a localized malignant melanoma in the mucosa of the upper jaw. PMID- 8416881 TI - Major salivary gland function in patients with radiation-induced xerostomia: flow rates and sialochemistry. AB - Radiation therapy for cancer of the head and neck region often causes salivary gland dysfunction and xerostomia. Several reports suggest that the submandibular/sublingual (SM/SL) glands may be less radiosensitive than the parotid. The purpose of this study was to evaluate differential radiation effects on the major salivary glands. Fifty patients with radiation-induced xerostomia were evaluated (33 males, 17 females; mean age 52.7). The average total tumor dose was 6034 cGy. Major salivary gland function was compared with that of 50 non irradiated controls. Salivary flow rates included unstimulated and stimulated flows of both the parotid and SM/SL glands. Sialochemical analyses included total protein, lysozyme, lactoferrin, sodium, chloride, and potassium. All four measures of salivary flow were significantly reduced in patients as compared to controls (p = .0001). Like the parotid, submandibular/sublingual gland dysfunction appears to be radiation dose- and field-dependent. Patients in the lowest radiation dose quartile (< or = 5000 cGy) had significantly increased salivary flow compared to those in the highest dose quartile (> or = 6800 cGy; p = .025). Glands that were partially irradiated were more likely to have some residual function than fully irradiated glands (p = .003). Lactoferrin content was increased in parotid saliva of radiation patients (p = .0001). Chloride content was significantly increased also (p = .0001). The SM/SL glands are clearly dysfunctional in post-irradiation xerostomia patients compared to controls, in terms of both flow rates and sialochemistry. PMID- 8416882 TI - Enhancement of radiation response on human carcinoma cells in culture by pentoxifylline. AB - PURPOSE: Pentoxifylline, a methylxanthine, has been shown to improve tumor tissue oxygenation in hypoxic murine tumors and prevent the late radiation injury of normal tissues in mice. The present cell culture studies were carried out to determine whether pentoxifylline would enhance the cellular radiation response of several human carcinoma cells in culture under various culture conditions. METHODS AND MATERIALS: Experiments were carried out with three human carcinoma cells grown in Eagle's MEM supplemented with 10% FCS. Cell survival was assayed by the colony forming ability of single plated cells to obtain dose-survival curves. RESULTS: Cells irradiated and exposed to pentoxifylline for 24 hr showed a significant enhancement of radiation-induced cytotoxicity. Pre-irradiation treatment with the drug did not change cellular response to radiation. The magnitude of radiation enhancement was dependent on the concentration of drug and exposure time up to the time corresponding to one cell cycle time. Among the three human carcinoma cells used (HeLa S-3 cervix carcinoma, MCF-7 breast carcinoma, and HT-29 colon carcinoma), HeLa S-3 cells were most susceptible to the combined treatment with the enhancement ratio of 1.4 at 1 mM. Among the derivatives of methylxanthines, caffeine and pentoxifylline were equally effective on a molar basis. The combined effect of pentoxifylline and purine nucleosides, including guanosine and deoxyguanosine, further enhanced the sensitizing effect of pentoxifylline. CONCLUSION: The present data along with other radiobiological effects of the drug suggest that pentoxifylline should be considered as a radiation enhancer for clinical radiotherapy. PMID- 8416883 TI - Lipid peroxidation does not appear to be a factor in late radiation injury of the cervical spinal cord of rats. AB - PURPOSE: We tested the role of lipid peroxidation in the demyelination and white matter necrosis associated with radiation injury of the central nervous system. METHODS AND MATERIALS: We irradiated the cervical spinal cords of female F344 rats (23 Gy) and assayed for the accumulation of the peroxidation byproducts malondialdehyde and hydroxyeicosatetraenoic acids, and for the consumption of the endogenous free radical scavengers vitamins E and C. We further tested the role of lipid peroxidation in radiation injury of the central nervous system by determining the sensitivity of the cervical spinal cord to radiation in rats on diets containing deficient, normal, and supplemental levels of the antioxidant vitamin E. Rats were placed on these diets at 4 weeks of age and irradiated (18.5 21.5 Gy) 16 weeks later. RESULTS: During the 5 months between irradiation and the onset of paralysis, no accumulation of peroxidation byproducts or consumption of endogenous scavengers was seen in the cervical spinal cords of the irradiated rats. The cervical spinal cords of some of the rats placed on the diets with deficient, normal, and supplemental levels of vitamin E were analyzed at the time of irradiation and contained 197 +/- 57, 501 +/- 19, and 717 +/- 35 pmol vitamin E/mg protein, respectively. Despite the statistical differences in these levels, the radiation sensitivity of the cervical spinal cord (ED50 for white matter necrosis) in rats receiving the three diets was not different (20.4, 20.7, and 20.6 Gy). CONCLUSION: These data do not support a role for free radical-induced lipid peroxidation in the white matter damage seen in radiation injury of the central nervous system. PMID- 8416884 TI - Combined modality treatment of the rhabdomyosarcoma R1H of the rat: influence of sequence of cisplatin and fractionated irradiation. AB - The effect of irradiation with 30 fractions of 2 Gy in 6 weeks combined with a single dose of 5mg/kg cisplatin was studied in the rhabdomyosarcoma R1H of the rat and tumor response and normal tissue toxicity were assessed for various combinations of radiotherapy and chemotherapy. Cisplatin was given 3 days before, during (after the 5th and after the 10th fraction), or 3 and 17 days after radiotherapy. Five of the 12 tumors treated with cisplatin injected 3 days after radiotherapy were locally controlled (42%; 95% confidence intervals: 14-70%) as compared to 0/10 for radiotherapy alone (0%; 0-21%; p < 0.02). A similar trend was found for cisplatin injected 17 days after irradiation (2/6 local controls; 33%; 0-71%). With cisplatin given 3 days before radiotherapy 1/13 local controls were observed (8%; 0-22%; p < 0.05 when tested vs. cisplatin 3 days after radiotherapy). Tumor cure was dependent upon tumor size at time of cisplatin administration with 7/9 small tumors (< 2 mm3) cured versus only 2/35 cures of larger tumors (> 2 mm3). By contrast, net growth delay and skin damage were the same for combined modality treatment and for irradiation alone. General toxicity as assessed by body weight change was significantly higher for animals treated with cisplatin before or after radiotherapy, whereas cisplatin during radiotherapy showed equal effects as compared to radiotherapy alone. Although for the rhabdomyosarcoma R1H of the rat the combined modality treatment was shown to be more effective than radiotherapy alone when cisplatin was applied after radiotherapy, general toxicity was also higher for this mode of treatment. PMID- 8416885 TI - Hyperthermia and radiation in advanced malignant melanoma. AB - Advanced melanoma (48 lesions in 40 patients) was treated with external microwave hyperthermia combined with radiation therapy between 1980-1988. Thirty-three lesions in 28 patients were evaluable for tumor response (mean age 64 years, 19 male, 9 female). Evaluable lesions received 13 to 66 Gy (mean 37 +/- 2 Gy) over 5 to 16 fractions (mean of 10) in 14 to 56 elapsed days (mean of 25). Tumor volume (pi/6*length*width*depth) was 62 +/- 16 cm3 (1-377 cm3). Hyperthermia was administered in 6.6 +/- 0.4 sessions (range 1-14), there were 3.2 +/- 0.4 thermal sensors per tumor (range 1-11) and 27 fields were treated twice-weekly (82%). Of the 33 evaluable lesions, 12 exhibited a complete response (36%), and 17 had a partial response (52%). Among the 12 complete responders four recurrences (33%) were observed at 8.6 +/- 1.4 months (median of 8.2 months). In superficial tumors with depth < or = 3 cm and with lateral dimensions within 2 cm of the boundaries of the microwave applicator, the complete response rate was 50% (11/22); whereas for patients with deeper tumors with depth > 3 cm, the complete response rate was 9% (1/11), p = 0.02. The minimal tumor thermal dose during the first hyperthermia treatment session correlated with response (t43min1 = 20 +/- 7 vs. 6 +/- 3 minEq43 degrees C for complete responders and noncomplete responders, respectively, p = 0.06); and 7 of 10 lesions that had t43min 1 > or = 8 minEq43 degrees C achieved a complete response whereas only 5 of 22 lesions (23%) that had t43min1 < 8 minEq43 degrees C did so (p = 0.01). However, neither the minimum tumor temperature during the first treatment, the median minimum tumor temperature over all treatment sessions nor the sum of minimum thermal dose over all treatment sessions correlated with tumor response. Twenty-three patients with 28 lesions died during follow-up (82%). The survival for complete responding patients with superficial lesions was 21.3 +/- 1.5 months compared to 4.5 +/- 0.5 months for patients with superficial lesions that did not experience a complete response (p = 0.0001). For patients with noncomplete responding lesions deeper than 3 cm survival was 4.4 +/- 0.6 months. Twenty lesions were treated without any skin reaction (42%, 20/48). Of the rest, 23 had erythema (48%, 23/48), seven had blistering (14%, 7/48) and one had ulceration of the skin.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8416886 TI - Is reseeding from the primary a plausible cause of node failure? AB - In a previous analysis of node failures in 1251 consecutive patients with node positive oropharyngeal and pharyngolaryngeal squamous cell carcinomas treated by external radiotherapy alone at the Institut Curie, the main reasons for patient exclusion were node recurrence associated with primary failure (N+T failures) and doses less than 55 Gy. These exclusions reduced the number of node failures from 399/1251 (32%) to 77/798 (10%). Multivariate analysis of node recurrence indicated that node size and fixity, treatment duration, and T stage of primary were significant (higher probability of isolated node failure for the T1-T2 primaries). In the present analysis, it is noted that 60% of the N+T failures were observed less than 1 month after the completion of the irradiation and, therefore, were not likely the result of reseeding from the primary tumor. When all 1251 patients were included in the analysis, the probability of nodal failure increased for larger nodes, T4 primaries, lower nodal doses, presence of contralateral node metastases, and nodal fixation to the surrounding structures. No influence of the primary site was found. Treatment duration was closely associated with total dose to the nodes. The best description of the data was obtained with a model including total dose and not treatment time. However, as in the previous analysis, the exclusion of low-dose (less than 55 Gy) treatments resulted in the loss of a significant dose-control relationship. We conclude that the majority of node failures is unlikely to result from reseeding from the primary tumor, and therefore should not be excluded from local-control analyses. From a more radiobiological point of view, the exclusion of palliative treatments is questionable when studying the effect of dose on local control. PMID- 8416887 TI - Enhancement of hyperthermia effect in vivo by amiloride and DIDS. AB - PURPOSE: Intracellular pH is regulated mainly by Na+/H+ antiport and Cl-/HCO3- exchange through the cell membrane. Amiloride (3,5-diamino-6-chloro-N (diaminomethylene)pyrazine carboxamide) is a diuretic drug that blocks Na+/H+ antiport and DIDS (4,4-diisothiocyanatostilbene-2,2'-disulfonic acid) is an inhibitor of Cl-/HCO3- exchange. We investigated the potency of these drugs to lower pHi and increase the thermosensitivity of tumors in vivo. MATERIALS AND METHODS: The cytocidal effect of heat in combination with drug effect in vivo was studied using the in vivo-in vitro clonogenic assay method and the tumor growth delay method with SCK tumors, a mammary adenocarcinoma, on the hind limbs of A/J mice. The effects of amiloride and DIDS on tumor pHi and high energy phosphate levels were investigated using 31P-NMR. RESULTS: We observed that amiloride or DIDS alone increased the effect of hyperthermia at 42.5 degrees C or 43.5 degrees C to suppress tumor growth. The thermosensitization was greater when the two drugs were combined. For example, hyperthermia at 43.5 degrees C alone resulted in a tumor growth delay of about 4 days. When 10 mg/kg amiloride or 25 mg/kg DIDS was injected prior to heating, the growth delay increased to about 6 days. When both drugs were injected prior to heating, a total growth delay of 8 days was obtained. In vivo-in vitro excision assays for cell survival demonstrated that these drugs enhanced the heat-induced tumor cell death. An i.p. injection of 10 mg/kg amiloride plus 25 mg/kg DIDS did not lower the tumor pHi over a 120 min interval. Heating the tumors at 42.5 degrees C for 1 hr significantly lowered the pHi and when the tumor-bearing mice were injected i.p with amiloride and DIDS, and the tumors were heated 1 hr later, the drop in pHi was greater relative to that by heating alone. Heating alone significantly lowered the tumor energy levels as indicated by PCr/Pi and beta-ATP/Pi ratios and an i.p. injection of 25 mg/kg amiloride prior to heating further reduced the energy status in the tumors. CONCLUSION: Amiloride or its analogs and DIDS may be useful in increasing the therapeutic efficacy of hyperthermia treatments by enhancing the reduction in tumor pHi. PMID- 8416888 TI - Leprosy, tuberculosis, and the new genetics. PMID- 8416889 TI - Genomic restriction map of the extremely thermophilic bacterium Thermus thermophilus HB8. AB - A physical map of the chromosome of the extremely thermophilic eubacterium Thermus thermophilus HB8 has been constructed by using pulsed-field gel electrophoresis techniques. A total of 26 cleavage sites for the rarely cutting restriction endonucleases HpaI, MunI, and NdeI were located on the genome. On the basis of the sizes of the restriction fragments generated, the genome size was estimated to be 1.74 Mbp, which is significantly smaller than the chromosomes of Escherichia coli and other mesophiles. Partial digestion experiments revealed the order of the six HpaI bands on the chromosome. Hybridization of isolated restriction fragments to pulsed-field gel-separated restriction digestions confirmed the deduced order of the HpaI fragments and allowed ordering and alignment of the NdeI and MunI fragments. In addition, 16 genes or gene clusters cloned from several different Thermus strains were located on the T. thermophilus HB8 chromosomal map by hybridization of gene probes to pulsed-field gel-resolved restriction digestions. PMID- 8416890 TI - Proteins antigenically related to methyl-accepting chemotaxis proteins of Escherichia coli detected in a wide range of bacterial species. AB - The four methyl-accepting chemotaxis proteins of Escherichia coli, often called transducers, are transmembrane receptor proteins that exhibit substantial identity among the sequences of their cytoplasmic domains. Thus, antiserum raised to one of these proteins recognizes the others and might be expected to recognize related proteins in other bacteria. We used antiserum raised to the transducer Trg in immunoblot experiments to probe a wide range of bacterial species for the presence of antigenically related proteins. Such proteins were detected in over 20 different species, representing 6 of the 11 eubacterial phyla defined by analysis of rRNA sequences as well as one archaebacterial group. Species containing proteins antigenically related to the transducers of E. coli included members of all four subdivisions of the phylum in which E. coli is placed, members of four of the six subdivisions of spirochetes, and two gliding bacteria. These observations provide substantial support for the notion that methyl accepting taxis proteins are widely distributed among the diversity of bacterial species. PMID- 8416891 TI - Characterization of the Rickettsia prowazekii pepA gene encoding leucine aminopeptidase. AB - The pepA gene, encoding a protein with leucine aminopeptidase activity, was isolated from Rickettsia prowazekii, an obligate intracellular parasitic bacterium. Nucleotide sequence analysis revealed an open reading frame of 1,502 bp that would encode a protein of 499 amino acids with a calculated molecular weight of 53,892, a size comparable to that of the protein produced in Escherichia coli minicells containing the rickettsial gene. Also, heat-stable leucine aminopeptidase activity was demonstrable in an E. coli peptidase deficient strain containing R. prowazekii pepA. Comparison of the amino acid sequence of the R. prowazekii PepA with the characterized leucine aminopeptidases from E. coli, Arabidopsis thaliana, and bovine eye lens revealed that 39.8, 34.9, and 34.0% of the residues were identical, respectively. Residues proposed to be part of the active site or involved in the binding of metal ions in the bovine metalloenzyme were all conserved in R. prowazekii PepA. However, despite the structural and enzymatic similarity to E. coli PepA, the R. prowazekii protein was unable to complement the cer site-specific, PepA-dependent recombination system found in E. coli that resolves ColE1-type plasmid multimers into their monomeric forms. PMID- 8416892 TI - Cloning, sequence, and expression of a chitinase gene from a marine bacterium, Altermonas sp. strain O-7. AB - The gene encoding an extracellular chitinase from marine Alteromonas sp. strain O 7 was cloned in Escherichia coli JM109 by using pUC18. The chitinase produced was not secreted into the growth medium but accumulated in the periplasmic space. A chitinase-positive clone of E. coli produced two chitinases with different molecular weights from a single chitinase gene. These proteins showed almost the same enzymatic properties as the native chitinase of Alteromonas sp. strain O-7. The N-terminal sequences of the two enzymes were identical. The nucleotide sequence of the 3,394-bp SphI-HindIII fragment that included the chitinase gene was determined. A single open reading frame was found to encode a protein consisting of 820 amino acids with a molecular weight of 87,341. A putative ribosome-binding site, promoter, and signal sequence were identified. The deduced amino acid sequence of the cloned chitinase showed sequence homology with chitinases A (33.4%) and B (15.3%) from Serratia marcescens. Regardless of origin, the enzymes of the two bacteria isolated from marine and terrestrial environments had high homology, suggesting that these organisms evolved from a common ancestor. PMID- 8416893 TI - Identification of a Saccharopolyspora erythraea gene required for the final hydroxylation step in erythromycin biosynthesis. AB - In analyzing the region of the Saccharopolyspora erythraea chromosome responsible for the biosynthesis of the macrolide antibiotic erythromycin, we identified a gene, designated eryK, located about 50 kb downstream of the erythromycin resistance gene, ermE. eryK encodes a 44-kDa protein which, on the basis of comparative analysis, belongs to the P450 monooxygenase family. An S. erythraea strain disrupted in eryK no longer produced erythromycin A but accumulated the B and D forms of the antibiotic, indicating that eryK is responsible for the C-12 hydroxylation of the macrolactone ring, one of the last steps in erythromycin biosynthesis. PMID- 8416894 TI - Detection of a nitrous oxide reductase structural gene in Rhizobium meliloti strains and its location on the nod megaplasmid of JJ1c10 and SU47. AB - The gene encoding a denitrification enzyme, nitrous oxide reductase (EC 1.7.99.6), in Rhizobium meliloti and other gram-negative bacteria was detected by hybridization to an internal 1.2-kb PstI fragment of the structural gene (nosZ) cloned from Pseudomonas stutzeri Zobell (W.G. Zumft, A. Viebrock-Sambale, and C. Braun, Eur. J. Biochem. 192:591-599, 1990). Homology to the probe was detected in the DNAs of two N2-fixing strains of P. stutzeri, two denitrifying Pseudomonas species, one Alcaligenes eutrophus strain, and 36 of 56 R. meliloti isolates tested. Except for two isolates of R. meliloti, all showed nitrous oxide reduction activity (Nos+). Therefore, at least part of the nosZ sequence appears to be conserved and widely distributed among denitrifiers, which include free living and symbiotic diazotrophs. By using Agrobacterium tumefaciens transconjugants harboring different megaplasmids of R. meliloti JJ1c10 and SU47, sequence homology with the nosZ probe was unequivocally located on the nod megaplasmid. A cosmid clone of JJ1c10 in which nosZ homology was mapped on a 4.2 kb BamHI fragment was selected. This cosmid, which conferred Nos+ activity to the R. meliloti wild-type strains ATCC 9930 and Balsac (Nos- and nondenitrifying, respectively) also restored Nos+ activity in the mutants of JJ1c10 and SU47 in which the 4.2-kb BamHI segment was deleted. Therefore, this segment contains sequences essential for nos gene expression in JJ1c10 and SU47 and thus confirms that the nod megaplasmid in JJ1c10 and SU47 which carries genes essential for symbiotic dinitrogen fixation also carries genes involved in the antagonistic process of denitrification. PMID- 8416895 TI - Effects of insertions and deletions in glnG (ntrC) of Escherichia coli on nitrogen regulator I-dependent DNA binding and transcriptional activation. AB - Phosphorylated nitrogen regulator I (NRI, also called NTRC), encoded by glnG (ntrC), stimulates transcription in Escherichia coli and other enteric bacteria from sites analogous to eukaryotic enhancers. We isolated 30 mutants, obtained without phenotypic selection, that have either a small insertion or deletion within glnG. Mutants were classified by the ability of NRI to repress the glnAp1 and glnL promoters and to activate two versions of the nitrogen-regulated glnAp2 promoter; each activity was measured in cells grown with three concentrations of NRI. The results were interpreted within the framework of the three-domain hypothesis of NRI structure. NRI is thought to contain a phosphorylated regulatory domain that controls binding of ATP, a central domain that hydrolyzes ATP and interacts with RNA polymerase, and a DNA-binding region of unknown extent. Our results suggest that the 70 amino acids from residue 400 to the carboxyl terminus constitute a DNA-binding domain that includes residues for specific contacts and dimerization. Our results also suggest that (i) transcription from glnAp2 without specific NRI-binding sites requires binding to sites with some similarity to the specific sites, and (ii) if an NRI variant can stimulate transcription, then increasing the concentration of NRI diminishes glnA expression for all mutants but one. PMID- 8416896 TI - Osmotic repression of anaerobic metabolic systems in Escherichia coli. AB - The influence of the osmolarity of the growth medium on anaerobic fermentation and nitrate respiratory pathways was analyzed. The levels of several enzymes, including formate dehydrogenase, hydrogenase, and nitrate reductase, plus a nickel uptake system were examined, as was the expression of the corresponding structural and regulatory genes. While some functions appear to be only moderately affected by an increase in osmolarity, others were found to vary considerably. An increase in the osmolarity of the medium inhibits both fermentation and anaerobic respiratory pathways, though in a more dramatic fashion for the former. fnr expression is affected by osmolarity, but the repression of anaerobic gene expression was shown to be independent of FNR regulatory protein, at least for hyd-17 and fdhF. This repression could be mediated by the intracellular concentration of potassium and is reversed by glycine betaine. PMID- 8416897 TI - Role of the carboxyl-terminal domain of TolA in protein import and integrity of the outer membrane. AB - The TolA protein is involved in maintaining the integrity of the outer membrane of Escherichia coli, as mutations in tolA cause the bacteria to become hypersensitive to detergents and certain antibiotics and to leak periplasmic proteins into the medium. This protein also is required for the group A colicins to exert their effects and for many of the filamentous single-stranded bacteriophage to infect the bacterial cell. TolA is a three-domain protein, with the amino-terminal domain anchoring it to the inner membrane. The helical second domain is proposed to span the periplasmic space to allow the carboxyl-terminal third domain to interact with the outer membrane. A plasmid that allowed the synthesis and transport of the carboxyl-terminal third domain into the periplasmic space was constructed. The presence of an excess of this domain in the periplasm of a wild-type cell resulted in an increased sensitivity to deoxycholate, the release of periplasmic alkaline phosphatase and RNase into the medium, and an increased tolerance to colicins E1, E2, E3, and A. There was no effect on the cells' response to colicin D, which depends on TonB instead of TolA for its action. The presence of the free carboxyl-terminal domain of TolA in the periplasm in a tolA null mutation did not restore the wild-type phenotype, suggesting that this domain must be part of the intact TolA molecule to perform its function. Our results are consistent with a model in which the carboxyl terminal domain of TolA interacts with components in the periplasm or on the inner surface of the outer membrane to function in maintaining the integrity of this membrane. PMID- 8416898 TI - Analysis of mRNA decay and rRNA processing in Escherichia coli multiple mutants carrying a deletion in RNase III. AB - RNase III is an endonuclease involved in processing both rRNA and certain mRNAs. To help determine whether RNase III (rnc) is required for general mRNA turnover in Escherichia coli, we have created a deletion-insertion mutation (delta rnc-38) in the structural gene. In addition, a series of multiple mutant strains containing deficiencies in RNase II (rnb-500), polynucleotide phosphorylase (pnp 7 or pnp-200), RNase E (rne-1 or rne-3071), and RNase III (delta rnc-38) were constructed. The delta rnc-38 single mutant was viable and led to the accumulation of 30S rRNA precursors, as has been previously observed with the rnc 105 allele (P. Gegenheimer, N. Watson, and D. Apirion, J. Biol. Chem. 252:3064 3073, 1977). In the multiple mutant strains, the presence of the delta rnc-38 allele resulted in the more rapid decay of pulse-labeled RNA but did not suppress conditional lethality, suggesting that the lethality associated with altered mRNA turnover may be due to the stabilization of specific mRNAs. In addition, these results indicate that RNase III is probably not required for general mRNA decay. Of particular interest was the observation that the delta rnc-38 rne-1 double mutant did not accumulate 30S rRNA precursors at 30 degrees C, while the delta rnc-38 rne-3071 double mutant did. Possible explanations of these results are discussed. PMID- 8416899 TI - A single amino acid substitution in elongation factor Tu disrupts interaction between the ternary complex and the ribosome. AB - Elongation factor Tu (EF-Tu).GTP has the primary function of promoting the efficient and correct interaction of aminoacyl-tRNA with the ribosome. Very little is known about the elements in EF-Tu involved in this interaction. We describe a mutant form of EF-Tu, isolated in Salmonella typhimurium, that causes a severe defect in the interaction of the ternary complex with the ribosome. The mutation causes the substitution of Val for Gly-280 in domain II of EF-Tu. The in vivo growth and translation phenotypes of strains harboring this mutation are indistinguishable from those of strains in which the same tuf gene is insertionally inactivated. Viable cells are not obtained when the other tuf gene is inactivated, showing that the mutant EF-Tu alone cannot support cell growth. We have confirmed, by partial protein sequencing, that the mutant EF-Tu is present in the cells. In vitro analysis of the natural mixture of wild-type and mutant EF-Tu allows us to identify the major defect of this mutant. Our data shows that the EF-Tu is homogeneous and competent with respect to guanine nucleotide binding and exchange, stimulation of nucleotide exchange by EF-Ts, and ternary complex formation with aminoacyl-tRNA. However various measures of translational efficiency show a significant reduction, which is associated with a defective interaction between the ribosome and the mutant EF-Tu.GTP.aminoacyl tRNA complex. In addition, the antibiotic kirromycin, which blocks translation by binding EF-Tu on the ribosome, fails to do so with this mutant EF-Tu, although it does form a complex with EF-Tu. Our results suggest that this region of domain II in EF-Tu has an important function and influences the binding of the ternary complex to the codon-programmed ribosome during protein synthesis. Models involving either a direct or an indirect effect of the mutation are discussed. PMID- 8416900 TI - Control of transcription of gal repressor and isorepressor genes in Escherichia coli. AB - Two regulatory proteins, Gal repressor and isorepressor, control the expression of the gal and mgl operons in Escherichia coli. The transcription start sites for galR and galS, the genes for the repressor and isorepressor, were determined by primer extension of in vivo transcripts. Study of the promoter-lacZ gene fusions introduced into the chromosome indicated that galS expression was elevated in cells in which the normal galS gene was interrupted, but not in cells in which the galR gene was deleted. When both genes were disrupted, galS expression was further elevated. Expression from the galS promoter was stimulated by the addition of D-fucose, repressed by glucose, and dependent on cyclic AMP receptor protein (CRP). Expression of a similar gene fusion of the galR promoter to lacZ was unregulated. Both galR and galS genes contain two potential operator sites (OE and OI) and a CRP-binding site. The arrangement of OE, OI, and the CRP binding site in the galS gene is analogous to the arrangement in the gal and mgl promoters, but the arrangement in galR is atypical. The increased concentration of the isorepressor when inducer is present may facilitate early shutoff of the isorepressor-regulated genes of the gal regulon when inducer (substrate) concentration falls. PMID- 8416901 TI - Osmotic regulation of rpoS-dependent genes in Escherichia coli. AB - The rpoS gene, which encodes a putative alternative sigma factor (sigma S), is essential for the expression of a variety of stationary-phase-induced genes as well as for stationary-phase-specific multiple-stress resistance. As previously shown for the otsA and otsB genes (R. Hengge-Aronis, W. Klein, R. Lange, M. Rimmele, and W. Boos, J. Bacteriol. 173:7918-7924, 1991), we demonstrate here that additional rpoS-controlled genes (bolA, csi-5) as well as at least 18 proteins on two-dimensional O'Farrell gels could be induced in growing cells by osmotic upshift via an rpoS-dependent mechanism. Also, rpoS-dependent thermotolerance and resistance against hydrogen peroxide could be osmotically stimulated. In contrast, the expression of glgS, while exhibiting strong stationary-phase induction, was only weakly increased by elevated osmolarity, and several rpoS-dependent proteins previously identified on two-dimensional gels were not osmotically induced. During osmotic induction of rpoS-dependent genes, rpoS transcription and the level of sigma S remained unchanged. We conclude that osmotically regulated genes represent a subfamily within the rpoS regulon that requires differential regulation in addition to that provided by sigma S. PMID- 8416902 TI - Identification and characterization of the Escherichia coli RecT protein, a protein encoded by the recE region that promotes renaturation of homologous single-stranded DNA. AB - Recombination of plasmid DNAs and recombination of bacteriophage lambda red mutants in recB recC sbcA Escherichia coli mutants, in which the recE region is expressed, do not require recA. The recE gene is known to encode exonuclease VIII (exoVIII), which is an ATP-independent exonuclease involved in the RecE pathway of recombination. A 33,000-molecular-weight (MW) protein was observed to be coexpressed with both exoVIII and a truncated version of exoVIII, pRac3 exo, when they were overproduced under the control of strong promoters. We have purified this 33,000-MW protein (p33) and demonstrated by protein sequence analysis that it is encoded by the same coding sequence that encodes the C-terminal 33,000-MW portion of exoVIII. p33 is expressed independently of exoVIII but is probably translated from the same mRNA. p33 was found to bind to single-stranded DNA and also to promote the renaturation of complementary single-stranded DNA. It appears that p33 is functionally analogous to the bacteriophage lambda beta protein, which may explain why RecE pathway recombination does not require recA. PMID- 8416903 TI - Host RecJ is required for growth of P22 erf bacteriophage. AB - Growth of bacteriophage P22 erf is known to require host RecA recombination function. We show that the RecA function is necessary but not sufficient to restore the plaque-forming ability of phage P22 erf; such mutant phage also requires host RecJ function. The residual efficiency of plaquing of P22 erf in a recJ background (0.03%) is completely abolished in recJ recB hosts (< 0.001%), suggesting that the RecBCD nuclease can provide an alternative function allowing phage growth. One tentative explanation is that circularization of P22 erf DNA mostly proceeds through the RecF pathway of recombination; however, less efficient circularization via the RecBCD pathway may also occur. In a recJ background, lysates obtained upon induction of an erf prophage show reduced yield (10%), suggesting that growth of P22 erf may require host RecJ in a step(s) other than circularization of phage DNA. PMID- 8416904 TI - Chemotactic signalling in Rhodobacter sphaeroides requires metabolism of attractants. AB - Rhodobacter sphaeroides showed chemotaxis towards L-alanine but not towards the analog 2-aminoisobutyrate. 2-Aminoisobutyrate and alanine were shown to share a common transport system, but 2-aminoisobutyrate was not metabolized. Chemotaxis towards alanine was inhibited by structurally unrelated metabolites, suggesting cross-inhibition by common metabolic intermediates. PMID- 8416905 TI - A genetic region downstream of the hydrogenase structural genes of Bradyrhizobium japonicum that is required for hydrogenase processing. AB - Deletion of a 2.9-kb chromosomal EcoRI fragment of DNA located 2.2 kb downstream from the end of the hydrogenase structural genes resulted in the complete loss of hydrogenase activity. The normal 65- and 35-kDa hydrogenase subunits were absent in the deletion mutants. Instead, two peptides of 66.5 and 41 kDa were identified in the mutants by use of anti-hydrogenase subunit-specific antibody. A hydrogenase structural gene mutant did not synthesize either the normal hydrogenase subunits or the larger peptides. Hydrogenase activity in the deletion mutants was complemented to near wild-type levels by plasmid pCF1, containing a 6.5-kb BglII fragment, and the 65- and 35-kDa hydrogenase subunits were also recovered in the mutants containing pCF1. PMID- 8416906 TI - Tryptophan biosynthesis genes trpEGC in the thermoacidophilic archaebacterium Sulfolobus solfataricus. AB - A DNA fragment containing the trpEGC gene cluster was isolated from the thermoacidophilic archaebacterium Sulfolobus solfataricus. The products of trpE, trpG, and trpC from S. solfataricus were compared to the homologous products from a eukaryote, a eubacterium, and two archaebacteria, namely, a methanogen and an extreme halophile. They appeared to be equally related to the proteins from Escherichia coli and Saccharomyces cerevisiae, the percentages of conserved amino acids being roughly the same as those measured when comparing the eubacterial and eukaryotic sequences directly. These percentages did not rise significantly when a comparison with the proteins from Haloferax volcanii was drawn, while a slightly closer relationship with the proteins from Methanococcus thermoautotrophicum was found. PMID- 8416907 TI - Mutational uncoupling of the transcriptional activation function of the TyrR protein of Escherichia coli K-12 from the repression function. AB - The tyrosine repressor (TyrR) protein of Escherichia coli can function either as a transcriptional enhancer or as a repressor. The structural basis for these opposite effects was analyzed in specific tyrR deletion mutants constructed in vitro. The functional behavior of the mutant TyrR proteins was evaluated in vivo by using single-copy lacZ reporter systems based on the mtr promoter (10-fold activation by wild-type TyrR protein, mediated by phenylalanine or tyrosine) or the aroF promoter (over 20-fold repression by wild-type TyrR protein, mediated by tyrosine). A mutant TyrR protein lacking amino acids 2 to 9 was completely devoid of transcriptional activation function. Five additional mutant TyrR proteins lacking progressively greater numbers of N-terminal amino acids were likewise activation defective. The mutant TyrR proteins lacking amino acid residues 2 to 9 or 2 to 19 were essentially identical to the wild-type TyrR protein in their ability to repress the aroF promoter. Three other TyrR mutant proteins, lacking up to 143 amino acid residues from the N-terminal end of the protein, retained the ability to repress the aroF promoter, to different extents, in a tyrosine dependent manner. PMID- 8416908 TI - Complete nucleotide sequence of a linear plasmid from Streptomyces clavuligerus and characterization of its RNA transcripts. AB - The complete nucleotide sequence of a small linear plasmid (pSCL1) from Streptomyces clavuligerus has been determined. This plasmid is 11,696 bp in length, has a 72% G+C content, and has approximately 900-bp inverted terminal repeat sequences. A comparison of the inverted terminal repeats of pSCL1 with those of a linear plasmid from S. rochei shows that the two terminal sequences have a high degree of similarity (approximately 70%). Several small inverted repeats found in the long terminal sequences of both plasmids are also conserved. An analysis of the sequence and codon preferences indicates that pSCL1 has seven or eight highly probable protein-coding open reading frames (ORFs). However, only two RNA species encoded by pSCL1 were detected in S. clavuligerus grown in liquid culture. The larger of these transcripts (900 nucleotides) corresponds to an ORF and is likely to be an mRNA for a protein similar to the KorA protein of pIJ101. The smaller transcript (460 nucleotides) does not correspond to any ORF; however, its 5' end is complementary to the 5' end of a predicted mRNA, suggesting that it may function as an antisense RNA. The larger of the two RNA species was present at a high level during the early stage of growth in liquid medium, and then its apparent rate of transcription decreased and remained at a lower level through the later stages; the level of the smaller RNA species remained relatively constant through all stages of growth. PMID- 8416909 TI - Role of Clp protease subunits in degradation of carbon starvation proteins in Escherichia coli. AB - When deprived of a carbon source, Escherichia coli induces the synthesis of a group of carbon starvation proteins. The degradation of proteins labeled during starvation was found to be an energy-dependent process which was inhibited by the addition of KCN and accelerated when cells were resupplied with a carbon source. The degradation of the starvation proteins did not require the ATP-dependent Lon protease or the energy-independent proteases protease I, protease IV, OmpT, and DegP. During starvation, mutants lacking either the ClpA or ClpP subunit of the ATP-dependent Clp protease showed a partial reduction in the degradation of starvation proteins. Strains lacking ClpP failed to increase degradation of starvation proteins when glucose was added to starving cells. The clpP mutants showed a competitive disadvantage compared with wild-type cells when exposed to repeated cycles of carbon starvation and growth. Surprisingly, the glucose stimulated, ClpP-dependent degradation of starvation proteins did not require either the ClpA or ClpB protein. The patterns of synthesis of starvation proteins were similar in clpP+ and clpP cells. The clpP mutants had reduced rates of degradation of certain starvation proteins in the membrane fraction when a carbon source was resupplied to the starved cells. PMID- 8416910 TI - Regulatory circuit for responses of nitrogen catabolic gene expression to the GLN3 and DAL80 proteins and nitrogen catabolite repression in Saccharomyces cerevisiae. AB - We demonstrate that expression of the UGA1, CAN1, GAP1, PUT1, PUT2, PUT4, and DAL4 genes is sensitive to nitrogen catabolite repression. The expression of all these genes, with the exception of UGA1 and PUT2, also required a functional GLN3 protein. In addition, GLN3 protein was required for expression of the DAL1, DAL2, DAL7, GDH1, and GDH2 genes. The UGA1, CAN1, GAP1, and DAL4 genes markedly increased their expression when the DAL80 locus, encoding a negative regulatory element, was disrupted. Expression of the GDH1, PUT1, PUT2, and PUT4 genes also responded to DAL80 disruption, but much more modestly. Expression of GLN1 and GDH2 exhibited parallel responses to the provision of asparagine and glutamine as nitrogen sources but did not follow the regulatory responses noted above for the nitrogen catabolic genes such as DAL5. Steady-state mRNA levels of both genes did not significantly decrease when glutamine was provided as nitrogen source but were lowered by the provision of asparagine. They also did not respond to disruption of DAL80. PMID- 8416911 TI - Turnover and recycling of the murein sacculus in oligopeptide permease-negative strains of Escherichia coli: indirect evidence for an alternative permease system and for a monolayered sacculus. AB - Turnover of murein in oligopeptide permease-negative Escherichia coli cells appeared to be minimal or nonexistent. In one strain in which it was possible to measure turnover during the first generation of chase, it was found that the rate of turnover was constant throughout a chase of three generations. This result suggests that an "inside-to-outside" mode of growth of the sacculus does not occur in E. coli. Turnover, though minimal, was significantly higher from cells labeled uniformly than from cells labeled only in the lateral wall, suggesting that a significant portion of the observed turnover is related to cell separation. Actually, turnover only appeared to be minimal in opp mutant strains. Tripeptides were being released by turnover at a rate of about 50% per generation and then were efficiently recycled. This suggests that in addition to opp, a low affinity uptake system for tripeptide derived from the sacculus may exist. PMID- 8416912 TI - Effect of glpT and glpD mutations on expression of the phoA gene in Escherichia coli. AB - In vivo 31P nuclear magnetic resonance analysis of Escherichia coli cells showed that the intracellular concentration of P(i) remained constant in wild-type and in a glpT mutant strain whether the cells were grown on excess (2 mM) P(i) or sn glycerol-3-phosphate as a phosphate source. The function of the phoA promoter (measured by beta-galactosidase activity in a phoA-lacZ fusion strain) was repressed when glpT+ cells were utilizing sn-glycerol-3-phosphate as the sole source of phosphate. These cells were devoid of alkaline phosphatase activity. However, the phoA promoter was fully active in a glpT mutant. These results indicated that the repression of the enzyme synthesis was not due to a variation in the level of cytoplasmic P(i) but was due to the P(i) excreted into the periplasm and/or to the medium. PMID- 8416913 TI - Membrane ATPase from the aceticlastic methanogen Methanothrix thermophila. AB - A new isolate of the aceticlastic methanogen Methanothrix thermophila utilizes only acetate as the sole carbon and energy source for methanogenesis (Y. Kamagata and E. Mikami, Int. J. Syst. Bacteriol. 41:191-196, 1991). ATPase activity in its membrane was found, and ATP hydrolysis activity in the pH range of 5.5 to 8.0 in the presence of Mg2+ was observed. It had maximum activity at around 70 degrees C and was specifically stimulated up to sixfold by 50 mM NaHSO3. The proton ATPase inhibitor N,N'-dicyclohexylcarbodiimide inhibited the membrane ATPase activity, but azide, a potent inhibitor of F0F1 ATPase (H(+)-translocating ATPase of oxidative phosphorylation), did not. Since the enzyme was tightly bound to the membranes and could not be solubilized with dilute buffer containing EDTA, the nonionic detergent nonanoyl-N-methylglucamide (0.5%) was used to solubilize it from the membranes. The purified ATPase complex in the presence of the detergent was also sensitive to N,N'-dicyclohexylcarbodiimide, and other properties were almost the same as those in the membrane-associated form. The purified enzyme revealed at least five kinds of subunits on a sodium dodecyl sulfate polyacrylamide gel, and their molecular masses were estimated to be 67, 52, 37, 28, and 22 kDa, respectively. The N-terminal amino acid sequences of the 67- and 52-kDa subunits had much higher similarity with those of the 64 (alpha)- and 50 (beta)-kDa subunits of the Methanosarcina barkeri ATPase and were also similar to those of the corresponding subunits of other archaeal ATPases. The alpha beta complex of the M. barkeri ATPase has ATP-hydrolyzing activity, suggesting that a catalytic part of the Methanothrix ATPase contains at least the 67- and 52-kDa subunits. PMID- 8416914 TI - The asiA gene of bacteriophage T4 codes for the anti-sigma 70 protein. AB - The anti-sigma 70 factor of bacteriophage T4 is a 10-kDa (10K) protein which inhibits the sigma 70-directed initiation of transcription by Escherichia coli RNA polymerase holoenzyme. We have partially purified the anti-sigma 70 factor and obtained the sequence of a C-terminal peptide of this protein. Using reverse genetics, we have identified, at the end of the lysis gene t and downstream of an as yet unassigned phage T4 early promoter, an open reading frame encoding a 90 amino-acid protein with a predicted molecular weight of 10,590. This protein has been overproduced in a phage T7 expression system and partially purified. It shows a strong inhibitory activity towards sigma 70-directed transcription (by RNA polymerase holoenzyme), whereas it has no significant effect on sigma 70 independent transcription (by RNA polymerase core enzyme). At high ionic strength, this inhibition is fully antagonized by the neutral detergent Triton X 100. Our results corroborate the initial observations on the properties of the phage T4 10K anti-sigma 70 factor, and we therefore propose that the gene which we call asiA, identified in the present study, corresponds to the gene encoding this T4 transcriptional inhibitor. PMID- 8416915 TI - Siderophore-mediated iron transport correlates with the presence of specific iron regulated proteins in the outer membrane of Rhizobium meliloti. AB - A universal chemical assay used to detect the production of siderophores in a range of Rhizobium strains showed that production is strain specific. Iron nutrition bioassays carried out on Rhizobium meliloti strains to determine cross utilization of their siderophores showed that R. meliloti 2011, 220-5, and 220-3 could each use the siderophores produced by the other two but not the siderophore produced by R. meliloti DM4 (and vice versa). Mutants of R. meliloti 2011 and 220 5 defective in siderophore production were isolated by Tn5-mob mutagenesis. The Tn5-mob-containing EcoRI fragment of mutant R. meliloti 220-5-1 was cloned into pUC19. By using this fragment as a probe, the presence of a homologous region was observed in R. meliloti 2011 and 220-3 but not in R. meliloti DM4. A complementing cosmid from a gene bank of R. meliloti 2011 was identified by using the same probe. Introduction of this cosmid into R. meliloti 102F34, a strain not producing a siderophore, resulted in the ability of this strain to produce a siderophore and also in the ability to utilize the siderophores produced by R. meliloti 2011, 220-5, and 220-3 but not the siderophore produced by R. meliloti DM4. A comparative analysis of the outer membrane proteins prepared from iron deficient cultures of R. meliloti 102F34 and 102F34 harboring the cosmid revealed the presence, in the latter, of a low-iron-induced outer membrane protein corresponding to a low-iron-induced protein in R. meliloti 2011, 220-5, and 220 3. This protein is not present in R. meliloti DM4. The results suggest that R. meliloti 2011, 220-5, and 220-3 produce siderophores that are identical or sufficiently similar in structure to be transported by the membrane transport system of each strain while also indicating that utilization of a particular siderophore is correlated with the presence of specific outer membrane proteins. PMID- 8416916 TI - Origins of DNA replication in metazoan chromosomes. AB - In metazoan chromosomes, initiation of DNA replication occurs primarily at specific sites (0.5-3 kb; the OBR) using the same replication fork mechanism favored by simple genomes. Nucleosome segregation is distributive. However, initiation events also occur at other sites randomly distributed throughout a larger initiation zone (8-55 kb). These nonspecific initiation events presumably occur at a lower frequency than those at the OBR. Although origins in native chromosomes appear to be genetically determined, ori sequences have been convincingly demonstrated only during programmed gene amplification. It seems likely that metazoan origins include both specific DNA sequences as well as chromatin structure and nuclear organization, something that may be difficult to reproduce with plasmid DNA. Definitive answers will come only with a functional assay for initiation that exhibits the sequence-specific characteristics observed in vivo. PMID- 8416917 TI - Regulation of elongation factor G GTPase activity by the ribosomal state. The effects of initiation factors and differentially bound tRNA, aminoacyl-tRNA, and peptidyl-tRNA. AB - The elongation factor G (EF-G) is responsible for the translocation of the ribosome along the mRNA chain. Under in vitro conditions, EF-G exhibits a very active uncoupled GTPase activity which is dependent on the presence of ribosomes and is modulated by mRNA-dependent binding of tRNA. In the absence of tRNA, uncoupled EF-G GTPase is inhibited by initiation factors IF1 and IF3, but not by initiation factor IF2. In the presence of N-fMet-tRNAfMet and poly(A,U,G) or in the presence of N-acetyl-Phe-tRNAPhe and poly(U), initiation factor IF2 causes an additional decrease of the uncoupled EF-G GTPase activity. This effect, however, is dependent on the presence of IF1 and IF3 and is obviously due to the mRNA- and initiation factor-dependent binding of N-fMet-tRNAfMet and N-acetyl-Phe-tRNAPhe, respectively, to the ribosomal P-site. Non-enzymatic binding of N-fMet-tRNAfMet and N-acetyl-Phe-tRNAPhe, however, causes a stimulation of uncoupled EF-G GTPase activity. The same effects are observed for Met-tRNA, Phe-tRNAPhe and uncharged tRNA. These findings are discussed in the light of the three-site model of the ribosome and the mechanism of translocation. PMID- 8416918 TI - Evidence for an alternative pathway for colchicine binding to tubulin, based on the binding kinetics of the constituent rings. AB - The kinetics of tropolone methyl ether binding to tubulin were measured by following the loss of colchicine binding capacity upon preincubation of tubulin with tropolone methyl ether. At 25 degrees C a bimolecular association rate constant of 2.7 (+/- 0.2) M-1 min-1 was determined, and from the temperature dependence an activation energy of 37 (+/- 8) kJ.mol-1 was calculated. By displacement experiments a dissociation rate constant of 2.9 (+/- 0.6) x 10(-2) min-1 was determined at 25 degrees C. The effect of 3',4',5' trimethoxyacetophenone (TMA) is 2-fold. TMA reduces the apparent association rate constant of colchicine, indicating that it equilibrates very rapidly and reversibly with the colchicine binding site. From this reduction the binding constant for TMA can be obtained. At 25 degrees C a value of 112 (+/- 13) M-1 is estimated. The binding of TMA is practically thermoneutral. Preincubation of tubulin with TMA over 30 min not only reduces the subsequent binding rate constant of colchicine but also the amplitude. This indicates that TMA also binds slowly in a second mode or site. Stopped-flow kinetic studies reveal that fast TMA binding competes for the initial binding of colchicine. From these results it is concluded that colchicine binds initially with its trimethoxybenzene ring and in a subsequent step with the tropolone ring. PMID- 8416919 TI - Structural elucidation of a variety of GalNAc-containing N-linked oligosaccharides from human urinary kallidinogenase. AB - Fifteen different structures of terminal GalNAc-containing N-linked oligosaccharides from human urinary kallidinogenase have been identified. These N linked oligosaccharides were mostly neutral, because sialic acid content was lower than 0.13 mol of sialic acid/mol of sugar chain, and sulfate was not detected. The oligosaccharides were released from pepsin-digested protein by glycoamidase A (from almond) digestion. The reducing ends of the oligosaccharide chains were aminated with a fluorescent reagent, 2-aminopyridine. The resulting mixture of pyridylamino derivatives of the oligosaccharides were separated by high performance liquid chromatography on an ODS-silica column, and 15 oligosaccharides were isolated. The structure of each oligosaccharide fraction was analyzed by two-dimensional sugar mapping, component sugar analysis, high resolution proton nuclear magnetic resonance and methylation analysis. It was found that each N-linked oligosaccharide associated with human urinary kallidinogenase contains unsubstituted GalNAc residues at the nonreducing terminal. These 15 oligosaccharides include 5 biantennary, 7 triantennary, and 3 tetraantennary oligosaccharides. PMID- 8416920 TI - Exchangeable GTP binding site of beta-tubulin. Identification of cysteine 12 as the major site of cross-linking by direct photoaffinity labeling. AB - After direct photoaffinity cross-linking of [3H]GTP to the beta-subunit of tubulin, followed by tryptic digestion and alkaline phosphatase treatment, we employed cis-diol-specific boronate gel chromatography and reversed-phase high pressure liquid chromatography to purify a peptide containing most of the covalently bound radioactivity. The sequence of this peptide corresponded to that of residues 3-19 of beta-tubulin. Residue 10 of the peptide, which is Cys-12 in beta-tubulin, could not be identified. The fast atom bombardment mass spectrum of this peptide showed the presence of a predominant species with a molecular mass of 2022 kDa (2021 kDa for the 12C variant), which is 255 Da greater than the molecular mass of the peptide. Fast atom bombardment collision-activated decomposition mass spectrometry analysis produced fragments which are consistent with the beta(3-19) peptide but having a unit of mass of 358 at position 12. Thermolysin digestion of the tryptic peptide restricted the cross-linking site to the 9-amino acid sequence, I(L)QAGQXGNQ. The molecular mass of this peptide was 1174 kDa, which is equal to the mass of the beta(7-15) peptide containing an extra group of mass 255. To explain the molecular masses of the two labeled peptides, which are 26 atomic mass units less than expected, a mechanism of photolabeling is proposed that involves opening of the guanine ring and loss of the C-6 carbonyl function as CO2. PMID- 8416921 TI - Ca(2+)-dependent stimulation of retinoblastoma gene product phosphorylation and p34cdc2 kinase activation in serum-stimulated human fibroblasts. AB - In serum-deprived human fibroblasts IMR-90 and WI-38 cells, the addition of fetal calf serum or basic fibroblast growth factor stimulates DNA synthesis in an extracellular Ca(2+)-dependent manner; the effect of serum on [3H]thymidine incorporation into DNA is 4-16-fold greater at 2.0 mM CaCl2 as compared with that at 0.03 mM CaCl2. By contrast, in SV40 virus-transformed WI-38 (SV-WI-38) cells DNA synthesis is essentially independent of the extracellular calcium concentration ([Ca]out) and serum growth factors. To explore the role of Ca2+ in mitogenic signal transduction through G1 to S phase cell cycle progression, we studied and compared the effect of [Ca]out on phosphorylation of RB protein, the product of a tumor suppressor retinoblastoma gene. In IMR-90 and WI-38 cells, serum or basic fibroblast growth factor induces an increase in the amount of hyperphosphorylated forms of RB protein in a manner strictly dependent on [Ca]out. In sharp contrast, in SV-WI-38 cells, the extent of RB phosphorylation is little affected by [Ca]out or the presence or absence of serum growth factors. In addition, potent calmodulin antagonists W-7 and calmidazolium, but not an inactive analogue W-12 or W-5, strongly inhibit serum-induced increases in DNA synthesis and RB phosphorylation in IMR-90 and WI-38 cells, whereas in SV-WI-38 cells, the inhibitory effect is much more limited. Under the same treatment conditions, we measured histone H1 kinase activity associated with anti-p34cdc2 immunoprecipitate and found that the serum-induced increase in p34cdc2 kinase activity is strongly dependent on [Ca]out and is potently inhibited by the active calmodulin antagonists in IMR-90 and WI-38 cells, but not in SV-WI-38 cells. In IMR-90 cells that have been incubated with serum in 0.03 mM [Ca]out for 24 h, restoration of [Ca]out to 2.0 mM results in initiation of DNA synthesis after 13 h and concomitant increases in RB phosphorylation and p34cdc2 histone H1 kinase activity. These results suggest that in human fibroblasts, Ca2+/calmodulin regulates the signaling cascade leading to cdc2 kinase activation, RB protein phosphorylation, and DNA synthesis and that this Ca(2+)-dependent regulation is abrogated in SV40-transformed cells. PMID- 8416922 TI - Characteristics of the operon for a putrescine transport system that maps at 19 minutes on the Escherichia coli chromosome. AB - The nucleotide sequence of the operon for the putrescine transport system that maps at 19 min on the Escherichia coli chromosome was determined. It contained four open reading frames encoding potF, -G, -H, and -I proteins. The potF protein (M(r) = 38,000) was inferred to be a putrescine-specific binding protein existing in a periplasmic fraction from the results of Western blot analysis of the cell fractions and from measurements of polyamine binding to the protein. The potG protein (M(r) = 45,000) had consensus amino acid sequences for the nucleotide binding site. The potH (M(r) = 35,000) and potI (M(r) = 31,000) proteins consisted of six putative transmembrane-spanning segments linked by hydrophilic segments of variable length as shown by hydropathy profiles. The spermidine putrescine transport system, which is mainly involved in spermidine transport, consisted of potA, -B, -C, and -D proteins (Furuchi, T., Kashiwagi, K., Kobayashi, H., and Igarashi, K. (1991) J. Biol. Chem. 266, 20928-20933). The homologies of the corresponding two proteins between those two systems, F and D, G and A, H and B, and I and C, were 35, 42, 37, and 36%, respectively. The initiation point of the transcription of the operon for the putrescine transport system was determined by primer extension and S1 nuclease mapping. Transcription started from the T residue located either 149 or 150 nucleotides upstream from the initiator AUG codon of potF protein mRNA. By making several subclones and a mutant lacking the potF gene, we showed that the expression of all four proteins was necessary for maximal putrescine transport activity. These results indicate that the putrescine transport system can also be defined as a bacterial periplasmic transport system. PMID- 8416923 TI - Cloning and characterization of the Saccharomyces cerevisiae gene encoding NAD dependent 5,10-methylenetetrahydrofolate dehydrogenase. AB - Saccharomyces cerevisiae possess a monofunctional, cytoplasmic NAD-dependent 5,10 methylenetetrahydrofolate (THF) dehydrogenase that converts 5,10-methylene-THF to 5,10-methenyl-THF (Barlowe, C. K., and Appling, D.R. (1990) Biochemistry 29, 7089 7094). We have now isolated the gene encoding this enzyme from a yeast genomic library using oligonucleotide probes based on internal peptide sequences from the purified protein. Nucleotide sequence analysis reveals a 320-amino acid open reading frame that contains both of the internal peptide sequences. The predicted molecular weight (36,236) is consistent with the estimated size (33,000-38,000) of the purified protein. Disruption of the chromosomal copy of the gene resulted in loss of NAD-dependent 5,10-methylene-THF dehydrogenase activity and led to a purine requirement in certain genetic backgrounds, confirming a role for this enzyme in the oxidation of cytoplasmic one-carbon units. A single gene was mapped to chromosome XI by hybridization to a yeast chromosomal blot. We propose MTD1 as the name for this gene. Northern analysis of total yeast RNA revealed a single transcript of approximately 1,100 nucleotides. Multiple transcription initiation sites were identified between 58 and 83 base pairs upstream of the start of translation. The amino acid sequences derived from the nucleic acid sequences of seven other methylene-THF dehydrogenases cloned to date have been found to be highly homologous. Although the predicted amino acid sequence of the yeast NAD dependent enzyme shows slight homology to the other sequences, it appears to be only distantly related to the other 5,10-methylene-THF dehydrogenases. PMID- 8416924 TI - ATP synthase of yeast mitochondria. Isolation and disruption of the ATP epsilon gene. AB - The nuclear gene encoding the subunit epsilon of the catalytic sector F1 of the yeast Saccharomyces cerevisiae ATP synthase was cloned and sequenced. Degenerated oligonucleotide primers were constructed from primary structure data. A part of the ATP epsilon gene was amplified by polymerase chain reaction from yeast genomic DNA. From the amplified DNA sequence a nondegenerated oligonucleotide probe was constructed and used for isolating a 2040-base pair EcoRI fragment bearing the whole gene. A 186-base pair open reading frame encoding a 62-amino acid polypeptide is described. The deduced amino acid sequence was one amino acid longer than the mature protein. A null mutant was constructed. The mutant strain was unable to grow on glycerol medium. The mutant mitochondria had no detectable oligomycin-sensitive ATPase activity. The catalytic sector appeared unstable during purification but F0-subunits were still bound to F1. The mutation promoted a highly oligomycin-sensitive uncoupling of the mitochondrial respiration rate. PMID- 8416925 TI - Cold-sensitive cytosolic 3,5,3'-triiodo-L-thyronine-binding protein and pyruvate kinase from human erythrocytes share similar regulatory properties of hormone binding by glycolytic intermediates. AB - Similar cold-sensitive properties, values of dissociation constants (Kd = 1 x 10( 10) M), and regulatory effectors were found for the cold-sensitive cytosolic 3,5,3'-triiodo-L-thyronine (L-T3)-binding protein (CTBP) and pyruvate kinase from human erythrocyte. Various metabolites of the blood cell were assayed for their effects on CTBP activity after heat and cold preincubation treatments. Among these compounds, five- and six-carbon phosphorylated sugars were effective in protecting the CTBP activity against cold inactivation, whereas only ATP and dATP blocked activation by heat treatments. The effects of fructose 1,6-bisphosphate, fructose 2,6-bisphosphate, and ATP were obtained at physiological concentrations. Three-carbon phosphorylated intermediates of glycolysis, ADP, AMP, cAMP, and GTP had no effect on cold and heat treatments. The monomer-tetramer interconversion of the enzyme was also regulated by fructose 1,6-bisphosphate and ATP. The association is under the control of fructose 1,6-bisphosphate, whereas the dissociation is under ATP control. This regulation may have physiological relevance since the hormone binds to the tetrameric form of the enzyme at a site other than the active site. PMID- 8416926 TI - Interrelations between assembly and secretion of recombinant human acetylcholinesterase. AB - Transport and secretion of recombinant human acetylcholinesterase (rHuAChE) were studied in transfected human 293 cells expressing either the oligomerized soluble enzyme or a monomeric mutant derivative in which Cys-580 was substituted by alanine (C580A). In cells expressing the wild-type enzyme, the gradual assembly of newly synthesized intracellular rHuAChE monomers into oligomers occurs within the endoplasmic reticulum. Secretion of mature wild-type enzyme into the medium is efficient and appears to be exclusive to multimeric forms. Consistently, intracellular oligomers, but not monomers, are endoglycosidase H-resistant, indicating that only oligomers undergo terminal glycosylation in the wild-type enzyme. In contrast, in cells expressing the dimerization-defective C580A mutant, newly synthesized rHuAChE monomers undergo terminal glycosylation and are secreted into the medium as efficiently as wild-type multimers. No significant difference between the intracellular transport rates of wild-type rHuAChE oligomers and mutant C580A monomers was revealed by probing with specific lectins. In both systems, transport and processing prior to the trans-Golgi galactosylation compartment appear to be rate-limiting, whereas the following passage to the cell surface is rapid. In conclusion, we suggest that in the presence of a free cysteine at the COOH terminus of the rHuAChE polypeptide, secretion of monomers is not effectuated, whereas in its absence, monomers are exported from the endoplasmic reticulum and are capable of traversing the entire secretory pathway. PMID- 8416927 TI - Post-translational cleavage of a histone H1-like protein in the sperm of Mytilus. AB - Starting with total RNA from spermatogenic cells of Mytilus trossulus and using random priming, we have cloned and sequenced the c-DNAs corresponding to two variants of the sperm-specific protein PLII* (phi 2B). DNA sequencing in conjunction with mass spectrometry and protein sequence data have allowed us to establish that of the three sperm-specific proteins present in the sperm of Mytilus (PL-II*(phi 2B), PL-III (phi 1), PL-IV (phi 3)), the first and the last one are the result of post-translational cleavage of a common precursor. This common precursor is a member of the histone H1 family, and it exhibits inter- and intraspecific microheterogeneity. PMID- 8416928 TI - Phe-Met-Arg-Phe-NH2 (FMRFa)-related peptides inhibit Na(+)-Ca2+ exchange in cardiac sarcolemma vesicles. AB - The molluscan cardioexcitatory tetrapeptide FMRF-amide (Phe-Met-Arg-Phe-NH2) and related peptides inhibit Na(+)-Ca2+ exchange in calf cardiac sarcolemma vesicles. FMRFa itself has a low inhibitory potency (IC50 = 750 microM) which completely resides in its COOH-terminal RFa portion. The physiologically active analog FLRFa is 10-fold more potent (IC50 = 60 microM). Two other substitutions of the Met2 in FMRFa, by either Ile or Lys increase inhibitory potency 7- and 50-fold, respectively. The inhibitory potency increases 300-500-fold if the NH2-terminal Phe1 in FMRFa is substituted by either Val or His (IC50 = 1-2 microM). The inhibitory activity of WnLRFa (IC50 = 40 microM) is lost when either the NH2 terminal amino group is acylated or the NH2-terminal Trp1 is deleted. These data suggest that the COOH-terminal portion is essential for the basic low potency inhibition of Na(+)-Ca2+ exchange, whereas the NH2-terminal portion is important for the potentiation of the inhibitory activity. Although the IC50 values of various peptides range widely (10(-6)-10(-3) M), all of them induce a complete inhibition. The dose-response pattern of the peptide-induced inhibition is identical for the Na(+)-Ca2+ exchange and its partial reaction, the Ca(2+)-Ca2+ exchange. The inhibitory effect is reversible and affects both Nai(or Cai) dependent 45Ca uptake and Nao-dependent 45Ca efflux, suggesting that the bidirectional movements of ions are altered. A mild pretreatment of vesicles with trypsin augments the Na(+)-Ca2+ exchange 1.5-fold but diminishes the inhibitory potency 3-4-fold, suggesting that the inhibition is mediated by an extravesicular membrane protein. The characteristics of the peptide-induced inhibition resemble the effect of opiates on Na(+)-Ca2+ exchange. FLRFa and dextrorphan (a non-opioid stereoisomer of an opiate agonist) are mutually exclusive inhibitors, suggesting that they may bind to the same site. This putative site lacks the pharmacological properties of opiate receptors and may be located either on the Na(+)-Ca2+ exchanger or at its vicinity. Endogenous analogs of FMRFa may regulate intracellular calcium via Na(+)-Ca2+ exchange. PMID- 8416929 TI - Structure-activity of deleted and substituted systemin, an 18-amino acid polypeptide inducer of plant defensive genes. AB - The primary structure-activity relationships of systemin, an 18-amino acid polypeptide from tomato leaves that regulates the expression of two wound inducible proteinase inhibitor genes in tomato and potato plants, were investigated. Analogs of systemin, the only example of a polypeptide signal from plants, were synthesized with progressive deletions of amino acids from both the NH2 terminus and COOH terminus and assayed in young excised tomato plants. All of the analogs exhibited severely decreased proteinase inhibitor-inducing activities, indicating that the entire 18-amino acid sequence is necessary for maximal activity. Deletion of the COOH-terminal Asp abolished inducing activity. Progressive replacement of each amino acid of the entire polypeptide with Ala revealed two regions, near residues Pro13, where Ala substitution reduced activity to less than 0.2%, and Thr17, which totally inactivated the analog. Other replacements with Ala had little or only moderate effects on activity. The two inactive analogs, des-Asp18 systemin and Ala17 systemin, were potent inhibitors of the inducing activity of the native systemin. These analogs, therefore, contain structural conformations sufficient for competition with systemin, but they are not competent for proteinase inhibitor gene induction. A synthetic COOH-terminal tetrapeptide, Met-Gln-Thr-Asp, retained low proteinase inhibitor inducing activity, but virtually any replacements with other amino acids either eliminated activity or reduced the activity to very low or nearly undetectable levels. These results indicate that residues near the COOH terminus of systemin are necessary for activity, possibly involving a phosphorylation at Thr17, and that other regions of the systemin sequence are important for interacting with a receptor(s) but are not sufficient to activate proteinase inhibitor gene expression. PMID- 8416930 TI - Identity of Saccharomyces cerevisiae tRNA(Trp) is not changed by an anticodon mutation that creates an amber suppressor. AB - A C35-->T mutation in an Escherichia coli tRNA(Trp) gene creates an amber suppressor which efficiently inserts glutamine in response to UAG codons in vivo (Soll, L., and Berg, P. (1969) Nature 223, 1340-1342). We have introduced the same change in a yeast tRNA(Trp) gene and demonstrated that the tRNA acts as an efficient amber suppressor in vivo. Amino acid sequence analyses were performed on chitinase produced by cells carrying the corresponding gene with a UAG codon at position 8 of the mature protein plus the mutant tRNA(Trp) gene. In contrast to comparable experiments with E. coli, tryptophan is inserted at a frequency > or = 80% by the yeast suppressor tRNA(Trp). Furthermore, in vitro charging experiments with the mutant tRNA(Trp) reveal no detectable increase in glutamine acceptor activity results from the C35-->T transition. The identity elements in E. coli tRNA(Gln) are well characterized (Jahn, M., Rogers, J., and Soll, D. (1991) Nature 352, 258-260). Sequence comparisons of the tRNA(Trp) and tRNA(Gln) molecules from E. coli reveal that the amber suppressor tRNA(Trp) has four of five identity elements required for glutaminyl-tRNA synthetase recognition. A similar comparison in the yeast system shows only two of the five potential identity elements are present. We conclude that, in spite of substantial structural similarities between yeast and E. coli aminoacyl-tRNA synthetases, fundamental differences can exist with regard to tRNA recognition. PMID- 8416931 TI - Isolation of vacuolar membrane H(+)-ATPase-deficient yeast mutants; the VMA5 and VMA4 genes are essential for assembly and activity of the vacuolar H(+)-ATPase. AB - The vacuolar membrane H(+)-ATPase of the yeast Saccharomyces cerevisiae is a multisubunit enzyme complex composed of an integral membrane V0 sector, and a peripherally associated V1 sector. Deletion of one of several structural genes for vacuolar H(+)-ATPase subunits was previously demonstrated to prevent proper assembly of the remaining V1 subunits onto the vacuolar membrane (Kane, P.M., Kuehn, M.C., Howald-Stevenson, I., and Stevens, T.H. (1992) J. Biol. Chem. 267, 447-454). A genetic screen was designed to identify new genes whose products were essential for the synthesis, assembly, and/or function of the yeast vacuolar H(+) ATPase. Mutants were identified based on phenotypes associated with vacuolar membrane H(+)-ATPase loss of function (vma), including an inability to grow on media buffered at neutral pH. Representatives in five complementation groups were identified, including four novel mutant vma5, vma21, vma22, and vma23, all of which were defective in vacuolar ATPase enzyme activity. We report here the characterization of two genes, VMA4 and VMA5, that encode peripheral subunits of the vacuolar H(+)-ATPase. We determined that VMA5 encodes the 42-kDa subunit of the vacuolar H(+)-ATPase. The VMA4 gene, originally described by Foury (Foury, F. (1990) J. Biol. Chem. 265, 18554-18560), was determined to encode the 27-kDa subunit of the purified yeast vacuolar H(+)-ATPase. Characterization of the vma5 and vma4 mutants revealed that the 42- and 27-kDa subunits are essential for the assembly of the peripheral membrane portion of the H(+)-ATPase onto the vacuolar membrane. PMID- 8416932 TI - Veratryl alcohol-dependent production of molecular oxygen by lignin peroxidase. AB - Veratryl alcohol- and H2O2-dependent production of oxygen by lignin peroxidase isozyme H2 (LiPH2) from Phanerochaete chrysosporium was investigated. Veratryl alcohol oxidation by LiPH2 decreased with increasing concentrations of H2O2 while oxygen evolution increased. The absorption spectrum of the LiPH2 in these experiments indicated that it was in the compound II state. We propose that O2 production results from the one electron oxidation of H2O2 by the veratryl alcohol cation radical to yield superoxide, as the addition of superoxide dismutase stimulated oxygen production. It has been reported previously that oxygen is consumed in reaction mixtures containing lignin peroxidase, H2O2, veratryl alcohol, and oxalate (Popp, J. L., Kalyanaraman, B., and Kirk, T.K. (1990) Biochemistry 29, 10475-10480). In the presence of oxalate, we observed oxygen consumption that was dependent on the H2O2 concentration. The ability of other methoxybenzenes to mediate oxygen production appeared to be related to their redox potential. It was concluded that cation radicals can oxidize H2O2 by one electron which results in the production of superoxide and the evolution of molecular oxygen. Thus, the rates of LiPH2-catalyzed O2 consumption or O2 production are dependent on the relative concentrations of H2O2 and oxalate. PMID- 8416933 TI - Labeling of cysteine 231 in acetylcholinesterase from Torpedo nobiliana by the active-site directed reagent, 1-bromo-2-[14C] pinacolone. Effects of 2,2' dipyridyl disulfide and other sulfhydryl reagents. AB - Acetylcholinesterase (AcChE, EC 3.1.1.7) was isolated from the electric organ of T. nobiliana and treated with the active-site-directed alkylating agent 1-bromo-2 [14C]pinacolone ([14C]BrPin), or with BrPin, which acts initially as a competitive inhibitor, Ki = 0.18 mM, and then inactivates the enzyme, k2 = 1.8 x 10(-4) s-1. AcChE aliquots were digested with trypsin and fractionated by reversed phase high performance liquid chromatography. Inactivation caused a decrease in one absorption peak and an increase in another, identified as the peptide beginning at Ala-222 and extending to Arg-242. 5-Trimethylammonio-2 pentanone, a competitive inhibitor, isosteric with acetylcholine, retarded the inactivation and decreased the quantity of labeled peptide. On sequencing, the 14C label was found associated with Cys-231. This was confirmed by comparison with synthesized S-pinacolonylcysteine, by study of effects of blocking the sequencing by o-phthalaldehyde, and by inactivation by 2,2'-dipyridyl disulfide (2-PDS), a thiol-specific reagent that acts initially as a competitive inhibitor, Ki = 0.042 mM, and then inactivates the enzyme, k2 = 5.0 x 10(-4) s-1. This is retarded by 5-trimethylammonio-2-pentanone, and prior inactivation by 2-PDS prevents subsequent reaction of [14C]BrPin in the active site. BrPin inactivates AcChEs from Electrophorus electricus and from human erythrocyte, but 2-PDS does not. Neither reagent inactivates butyrylcholinesterases from human and horse serum. PMID- 8416934 TI - Active sites of the cytochrome p450cam (CYP101) F87W and F87A mutants. Evidence for significant structural reorganization without alteration of catalytic regiospecificity. AB - Ferricyanide oxidation of the aryl-iron complexes formed by the reaction of cytochrome P450 enzymes with arylhydrazines causes in situ migration of the aryl group from the iron to the porphyrin nitrogen atoms. The regiochemistry of this migration, defined by the ratio of the four possible N-arylprotoporphyrin IX isomers, provides a method for mapping the topologies of cytochrome P450 active sites. The method has been validated by using it to examine the active site of cytochrome P450cam (CYP101), for which a crystal structure is available. In agreement with the crystal structure, reaction with phenylhydrazine gives a 5:25:70 ratio of the NA:NC:ND (subscript indicates pyrrole ring) N phenylprotoporphyrin IX isomers. Naphthylhydrazine, however, yields exclusively the NC regioisomer and 4-(phenyl)phenylhydrazine the NA:NC:ND isomers in a 14:40:46 ratio. These isomer ratio differences are readily explained by topological differences between the upper and lower reaches of the active site. Having validated the aryl-iron shift as a topological probe, we used it to investigate the structural changes caused by mutation of Phe-87, a residue that provides the ceiling over pyrrole ring D in the crystal structure of cytochrome P450cam. Mutation of Phe-87 to a tryptophan causes no detectable change in the regiochemistry of camphor hydroxylation and only minor changes in the N-aryl isomer ratios. However, mutation of Phe-87 to an alanine, which was expected to open up the region above pyrrole ring D, severely decreased the proportion of the ND in favor of the NA isomer. Less rather than more space is therefore available over pyrrole ring D in the F87A mutant despite the fact that the regiochemistry of camphor hydroxylation remains unchanged. These results provide evidence for significant structural reorganization in the upper regions of the substrate binding site without alteration of the camphor hydroxylation regiospecificity in the F87A mutant. PMID- 8416935 TI - Reserpine binding to a vesicular amine transporter expressed in Chinese hamster ovary fibroblasts. AB - The potent antihypertensive drug reserpine inhibits the transport of biogenic amines into adrenal chromaffin granules and synaptic vesicles. Reserpine acts by binding almost irreversibly to the vesicular amine transporter, and this interaction has been used both to study the mechanism of transport and to purify the protein responsible. Recent isolation of a cDNA for the rat chromaffin granule amine transporter (CGAT) by selection in the neurotoxin 1-methyl-4 phenylpyridinium now permits an analysis of the interaction with reserpine at a molecular level. Using membranes from stable transformants expressing the transporter, we show that reserpine binds specifically and quantitatively to CGAT. As with the native protein in bovine chromaffin granules, a pH gradient accelerates reserpine binding, and amine substrates compete for binding with reserpine. However, 1-methyl-4-phenylpyridinium and tetrabenazine, the other principal inhibitor of vesicular amine transport, compete very poorly with reserpine for binding, suggesting that they interact with CGAT at distinct sites. PMID- 8416936 TI - Potentiation of calcium- and caffeine-induced calcium release by cyclic ADP ribose. AB - Cyclic ADP-ribose (cADPR) is a naturally occurring metabolite of NAD+ that is as potent as inositol 1,4,5-trisphosphate (IP3) in mobilizing Ca2+ in sea urchin eggs. Previous pharmacological evidence suggests that cADPR acts through a system similar to the Ca(2+)-induced Ca2+ release (CICR). Here I showed that in the presence of low concentrations of cADPR addition of Ca2+ to egg homogenates stimulated further release of Ca2+ in a concentration-dependent manner. In the absence of cADPR, no induced release was seen, and the added Ca2+ was, instead, sequestered by a thapsigargin-sensitive transport system. High concentrations of strontium (> 50 microM) could also induce Ca2+ release. The effective concentrations of Sr2+, however, were reduced 10-20-fold in the presence of low concentrations of cADPR. Barium, at up to 0.4 mM, did not stimulate Ca2+ release with or without cADPR. The potentiation between divalent cations and cADPR was mutual since the Ca2+ releasing activity of cADPR was also increased in the presence of strontium. Ionomycin and thapsigargin both released Ca2+ but neither potentiated Ca2+ release induced by divalent cations. Caffeine also released Ca2+ in a concentration-dependent manner, and its potency was greatly increased by low concentrations of cADPR, while no such simulation was seen with IP3. Conversely, low concentrations of caffeine that were not sufficient to release Ca2+ increased the effectiveness of cADPR 10-fold. Isocaffeine, an isomer of caffeine, was four to five times less effective, demonstrating the specificity of the caffeine effect. These results suggest that cADPR can function as an endogenous regulator of CICR in eggs. PMID- 8416937 TI - Aspartic acid 320 is required for optimal activity of rat pancreatic cholesterol esterase. AB - The acidic amino acid residue required for the catalytic activity of rat pancreatic cholesterol esterase has been identified in this study by sequence comparison with other serine esterases and by site-directed mutagenesis experiments. The sequence comparison studies identified 3 acidic residues in homologous domains between cholesterol esterase, acetylcholinesterase, cholinesterase, and Geotrichum candida lipase that may potentially be the catalytic acidic residue in these proteins. The role of Glu78, Asp79, and Asp320 in the catalytic activity of rat cholesterol esterase was then addressed by mutagenesis and expression of the cDNA. Results showed that replacement of Glu78 or Asp79 with alanine has no effect on the ability of the cholesterol esterase to hydrolyze the artificial water-soluble substrate p-nitrophenyl butyrate. In contrast, the Asp320-->Ala320 substitution abolished the enzyme activity of the cholesterol esterase. The specific requirement of Asp320 for optimal enzyme activity was demonstrated by substitution of the aspartic acid with glutamic acid, thus retaining the charge unit at this position. The Asp320-->Glu320 substitution resulted in an enzyme that displayed normal interaction with bile salt. However, catalytic activity of this mutagenized protein was reduced by approximately 50%. These results strongly suggested that aspartic acid 320 is an important component of the catalytic triad of pancreatic cholesterol esterase. The specific requirement of aspartic acid, instead of glutamic acid, for optimal activity is different from that of other members of the serine esterase gene family. PMID- 8416938 TI - Studies of the DNA binding properties of histone H4 amino terminus. Thermal denaturation studies reveal that acetylation markedly reduces the binding constant of the H4 "tail" to DNA. AB - The effect of acetylation on the DNA binding properties of the rigidly conserved histone H4 amino-terminal tail has been studied in detail using the technique of thermal denaturation. The quantitative DNA-binding parameters for both the non- and fully acetylated H4 amino terminus have been determined from thermal denaturation data for complexes of the peptides bound to mixed sequence 146-base pair DNA. We find that under dilute buffer conditions (5 mM Tris-HCl) the binding constant for the non-acetylated peptide to double-stranded DNA is 5 x 10(11) M-1 and that acetylation of lysine residues in the peptide reduces the binding constant to 1 x 10(5) M-1. The dramatic differences observed in the binding constants for the non- and fully acetylated peptides are probably due to the effect of acetylation on the even distribution of positively charged residues in the H4 amino terminus. In other experiments, the binding of both peptides to a 30 base pair oligonucleotide has been studied in solution with varying concentrations of sodium, magnesium, and phosphate ions. These experiments demonstrate that both magnesium and phosphate ions have strong effects on the binding of the H4 tail to DNA, especially weakening the binding of the acetylated peptide. For instance, the dissociation of the non-acetylated peptide from DNA requires 6 mM magnesium, yet the binding of the acetylated peptide is abolished in only 30 microM magnesium. The modulation of the DNA binding interactions of the H4 amino terminus by physiologically relevant ionic conditions, in addition to the effect of acetylation, can be important in the regulation of chromatin structure and function. PMID- 8416939 TI - Heteronuclear NMR analysis of unsaturated fatty acids in poly(3 hydroxyalkanoates). Study of beta-oxidation in Pseudomonas putida. AB - Poly(3-hydroxyalkanoates) (PHAs) were isolated from Pseudomonas putida KT2442 cultivated on petroselenic acid, oleic acid, and linoleic acid to study beta oxidation of unsaturated fatty acids. Both saturated and unsaturated medium chain length 3-hydroxy fatty acids were found to be constituents of these polymers. With the aid of proton-detected multiple quantum coherence and proton-detected multiple bond coherence NMR spectra the structures of the unsaturated monomers were identified as 3-hydroxy-5-cis-tetradecanoate for PHA produced on oleic acid, and 3-hydroxy-6-cis-dodecanoate and 3-hydroxy-5-cis-8-cis-tetradecadienoate for PHA produced on linoleic acid. The identified structures, which are derived from fatty acid degradation intermediates, indicate a degradation of oleic acid via the enoyl-CoA isomerase-dependent route and a degradation of linoleic acid via the dienoyl-CoA reductase-dependent route. PMID- 8416940 TI - Characterization and developmental expression of a novel sulfotransferase for the biosynthesis of sulfoglucuronyl glycolipids in the nervous system. AB - Sulfoglucuronyl glycolipids (SGGLs) are temporally and spatially regulated molecules in the developing nervous system. A novel sulfotransferase (ST) from rat brain which catalyzes the terminal step in the biosynthesis in vitro of SGGLs is described. The enzyme catalyzes a transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to a hydroxyl group on carbon 3 of the terminal glucuronyl residue in IV3 beta-glucuronyl neolactotetraosylceramide (GlcAnLcOse4Cer) and VI3 beta-glucuronyl neolactohexaosylceramide (GlcAnLcOse6Cer) to form 3-sulfated glucuronyl glycolipids. The enzyme is highly specific for glucuronylglycolipids (GGLs) and requires the free-COOH group of the terminal glucuronic acid for reactivity. GGL:ST present in the microsomal membranes requires Mn2+ ions and a nonionic detergent, Triton X-100 for activity. The optimal pH is 7.2 with Tris HCl buffer and Km values were 7 microM for 3'-phosphoadenosine 5'-phosphosulfate and 29 microM for GlcAnLcOse4Cer. GGL:ST was shown to be different from previously well studied galactocerebroside:sulfotransferase for the synthesis of myelin membrane-specific lipid sulfatide. This conclusion was based upon several criteria, i.e. including different requirements of incubation conditions for maximal activity, substrate competition experiments, different effects of heat, dithiothreitol, NaCl, and pyridoxal phosphate, as well as different profiles of expression of activity during development of the nervous tissues. The two enzymes were also partially resolved on a pyridoxal phosphate-ligated agarose column. Studies on the developmental expression of the GGL:ST in the rat cerebral cortex and cerebellum showed that it is not a regulatory enzyme controlling the expression of SGGLs in these neural tissues. PMID- 8416941 TI - Oxidative inactivation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase and subunit cross-linking involve different dithiol/disulfide centers. AB - Rat liver microsomal 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase (HMGR) is extremely sensitive to oxidative inactivation by low concentrations (micromolar) of glutathione disulfide (GSSG) even in the presence of millimolar concentrations of glutathione (GSH). Inactivation involves the formation of an intramolecular protein-SS-protein disulfide in thiol/disulfide redox equilibrium with the reduced, active enzyme (Cappel, R.E., and Gilbert, H.F. (1988) J. Biol. Chem. 263, 12204-12212). In the absence of dithiothreitol, HMGR oxidation has been previously shown to cross-link the microsomal enzyme into a covalent dimer (Ness, G.C., McCreery, M.J., Sample, C.E., Smith, M., and Pendelton, L.C. (1985) J. Biol. Chem. 260, 12391-12393). Examination of the extent of HMGR cross-linking and residual HMGR activity in microsomes equilibrated with glutathione redox buffers establishes that inactivation and cross-linking result from oxidation of different dithiol pairs. The thiol/disulfide oxidation potential (K(ox)) for the oxidative inactivation of HMGR, E(SH)2,active + GSSG<-->E(S-S)inactive + 2 GSH, is 0.67 +/- 0.07 M. However, the equilibrium constant for HMGR cross-linking, E(SH)2,monomer + GSS<-->E(S-S)dimer + 2 GSH, is 0.19 +/- 0.02 M, significantly lower than that for inactivation (p < 0.001). Because of the significantly different oxidation potentials and the lack of a linear relationship between cross-linking and inactivation, the two processes must involve two different sets of vicinal dithiols. HMGR becomes 5-10-fold more difficult to oxidize in the presence of saturating levels of the substrate, HMG-CoA. Both inactivation and cross-linking exhibit significantly lower oxidation potentials in the presence of this substrate, 0.072 +/- 0.01 and 0.047 +/- 0.007 M, respectively. The decrease in oxidation potential caused by substrate binding is observed for both inactivation and cross-linking, showing that both processes are affected by the binding of substrate to the enzyme. The dithiols involved in HMGR subunit cross linking are 2-3-fold more difficult to oxidize than the dithiols that affect the enzyme activity. Thus, the observation of partial cross-linking of HMGR in vivo would imply that conditions are sufficiently oxidizing to result in significant enzyme inactivation. The extreme thermodynamic sensitivity of HMGR to oxidative inactivation and cross-linking in glutathione redox buffers that span the physiological redox state implies that thiol/disulfide redox state changes could provide a mechanism for regulating the activity and/or stability of this enzyme. PMID- 8416942 TI - Purification and characterization of kumamolysin, a novel thermostable pepstatin insensitive carboxyl proteinase from Bacillus novosp. MN-32. AB - We have found a novel type of thermostable, pepstatin-insensitive carboxyl proteinase in the culture filtrate of Bacillus novosp. MN-32. The carboxyl proteinase, which was named kumamolysin, was purified about 8,300-fold by column chromatography including DEAE-Sepharose CL-6B, Sephadex G-100, and TSKgel DEAE 5PW. The purified kumamolysin gave a single band corresponding to 41 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass of kumamolysin was estimated to be 40 kDa by gel filtration. The isoelectric point of kumamolysin was estimated to be pH 3.5 by isoelectric focusing. Kumamolysin has maximum proteolytic activity at 70 degrees C and at pH 3.0. Kumamolysin specifically hydrolyzed the Leu15-Tyr16 peptide bond in oxidized insulin B-chain (Km = 9.0 x 10(-5) M, Kcat = 71 s-1; at pH 3.0, 30 degrees C), and additional cleavage at Phe25-Tyr26 was detected at a considerably lower rate. Kumamolysin is insensitive to the known carboxyl proteinase inhibitors pepstatin, diazoacetyl-DL norleucine methyl ester, and 1,2-epoxy-3-(p-nitrophenoxy)propane. Kumamolysin has no similarity to the thermostable acid protease thermopsin from Sulfolobus acidocaldarius (Lin, X.-L., and Tang, J. (1990) J. Biol. Chem. 265, 1490-1495). Thus, the substrate specificity, the inhibitor sensitivity, the molecular mass, and the thermostability all suggest that kumamolysin is a novel type of carboxyl proteinase. PMID- 8416943 TI - Roles of phospholipase C and Ca(2+)-ATPase in calcium responses of single, fibrinogen-bound platelets. AB - Fura-2-loaded platelets were immobilized on fibrinogen, and cytosolic free calcium concentration ([Ca2+]i) was measured by video imaging of the fluorescence signal. In the immobilized, single platelets, ADP and low doses of thrombin evoked repetitive spikes of [Ca2+]i, whereas higher thrombin concentrations gave elevated plateaus in [Ca2+]i. Stimulation of the cells with thrombin after the addition of ADP changed the frequency of spiking, but not the maximal levels of [Ca2+]i reached. In suspensions of platelets, ADP and low doses of thrombin evoked transient elevation of myo-inositol phosphates, suggesting that the initiation of spiking was due to stimulation of phospholipase C. In platelet suspensions, the Ca(2+)-ATPase inhibitor thapsigargin (TG) evoked a gradual rise in [Ca2+]i, which was potentiated by preactivation of the platelets and inhibited by prostaglandin E1 and nitroprusside. In single platelets, TG induced a sudden increase in [Ca2+]i after a long but variable delay, followed by a phase of slow oscillations. The effects of preactivation with ADP were 2-fold: the delay time before the response to TG was shortened and the maximal level of [Ca2+]i reached with TG was higher than the level of the preceding Ca2+ spikes. Apparently, Ca2+ responses induced by the inhibition of Ca(2+)-ATPases are potentiated by prior elevation of [Ca2+]i and reduced by substances that inhibit agonist-evoked increases in [Ca2+]i. The data point to a mechanism of Ca(2+)-induced Ca2+ release and the presence of TG-sensitive and TG-insensitive Ca2+ stores in platelets. Rapid spiking may involve both pools, whereas the latter alone may account for the slow TG-evoked oscillations. PMID- 8416944 TI - Truncated forms of mannose-binding protein multimerize and bind to mannose-rich Salmonella montevideo but fail to activate complement in vitro. AB - Human serum (MBP) and human recombinant (rMBP) mannose-binding protein bind to mannose-rich, serum-resistant Salmonella montevideo (SH5770), enhance C3 deposition, and render the organisms serum-sensitive. We investigated structural features of MBP necessary for this effect. MBP has a cysteine-rich amino-terminal region, a collagen-like region, and a carboxyl-terminal carbohydrate-recognition domain. We prepared carbohydrate-recognition domains lacking the other two domains either by deletion mutagenesis (delta MBP, 16 kDa) or by collagenase digestion of whole rMBP (cdMBP, 16-18 kDa). Whole and truncated MBP were detected on immunoblot by specific monoclonal antibodies that recognize both bound and free MBP. rMBP enhanced C3 deposition on SH5770 8-fold, while cdMBP and delta MBP did not increase C3 deposition over control levels. All forms of MBP bound to SH5770 by enzyme-linked immunosorbent assay and by measuring binding of radiolabeled whole and truncated MBP. Binding by 125I-delta MBP was inhibited by mannan and by MBP. Thus failure of truncated MBPs to enhance C3 deposition was not due to failure to specifically bind carbohydrate residues. To determine the molecular form of truncated MBP in nondenaturing conditions, 125I-delta MBP was centrifuged through a 5-20% sucrose-density gradient. The peak of 125I-delta MBP sedimented to estimated S20,w 2.01, but larger multimers also were present. Multimers bound SH5770 with higher affinity than monomers. We conclude that carbohydrate-recognition regions of MBP produced by collagenase digestion or by deletion mutagenesis are sufficient for ligand binding. However, the collagen like region is necessary for MBP enhancement of C3 deposition on SH5770. PMID- 8416945 TI - A cryptic, microsomal-type arachidonate 12-lipoxygenase is tonically inactivated by oxidation-reduction conditions in cultured epithelial cells. AB - Cultured ovine tracheal epithelial cells converted arachidonic acid to prostaglandin E2 (PGE2), but microsome-containing subcellular fractions prepared from these cells under calcium-free conditions converted arachidonic acid to PGE2 and to 12-hydroxyeicosatetraenoic acid (12-HETE) at a high rate (2-4 nmol/mg of protein/15 min). Identification of the membrane-bound 12-HETE-forming activity as a 12-lipoxygenase included 12S-stereospecificity of product formation and trapping of 12-hydroperoxyeicosatetraenoic acid as a reaction product. The 12 lipoxygenase activity was extracted from cell membranes only with detergent (1% Triton X-100), and the activity (membrane-bound or detergent-solubilized) was completely inactivated by mixing with the cytosol-containing subcellular fraction. The inhibitory effect of the cytosolic fraction was reversed by treating the cytosol with GSH-depleting agents (2-cyclohexene-1-one or N ethylmaleimide) or by mixing it with lipid hydroperoxide (13 hydroperoxyoctadecadienoic acid) at a concentration that had little direct effect on enzyme activity. Inhibition of 12-lipoxygenase activity could also be achieved by treatment of enzyme preparations with GSH at levels (0.1-10 mM) found in epithelial cell cytosol. In addition, treatment of cultured epithelial cells with a GSH-depleting agent (buthionine sulfoximine) and lipid hydroperoxide restored cellular 12-lipoxygenase activity. Little or no detectable 12-lipoxygenase activity was found in freshly isolated ovine tracheal epithelial cells, but the cytosolic 12-lipoxygenase found in freshly isolated bovine tracheal epithelial cells was relatively insensitive to regulation by GSH or lipid hydroperoxide. These observations indicate that a 12-lipoxygenase is expressed in a cryptic, microsomal-type form in primary-culture epithelial cells and that this form of the enzyme may be selectively regulated by changes in cellular oxidation reduction conditions dependent on cytosolic levels of GSH versus lipid hydroperoxide. PMID- 8416946 TI - Structural requirements for high affinity binding of complex ligands by the macrophage mannose receptor. AB - The mannose receptor of macrophage and hepatic endothelial cells discriminates between endogenous and exogenous sugar-bearing structures. Previous competition studies have indicated that the receptor binds the monosaccharides mannose, fucose, and N-acetylglucosamine but displays much higher affinity for multivalent oligosaccharides, such as those found on the surface of potentially pathogenic microorganisms. The hydrodynamic properties of the receptor have been examined, revealing that the receptor is a monomer. This result suggests that multiple carbohydrate recognition domains (CRDs) in the extracellular domain of a single receptor polypeptide cooperate to achieve high affinity binding of complex ligands. In order to determine the importance of individual CRDs, properties of receptor segments containing groups of CRDs expressed in insect cells have been examined. The results indicate that two of the CRDs (4 and 5) form a protease resistant, ligand-binding core but that five CRDs in tandem (4-8) are required to match the affinity of the intact receptor for yeast mannan. A consequence of the organization of the receptor is that both valency and geometry of glycoconjugates are important determinants of binding affinity. PMID- 8416947 TI - Alteration of Ca2+ permeability and sensitivity to Mg2+ and channel blockers by a single amino acid substitution in the N-methyl-D-aspartate receptor. AB - The N-methyl-D-aspartate (NMDA) receptor plays an important role in glutamate mediated neuronal plasticity and neurotoxicity in the central nervous system. This receptor is composed of a fundamental subunit (NMDAR1) and its potentiating subunits (NMDAR2A-NMDAR2D). The NMDA receptor is distinct from other glutamate receptor channels because of its high Ca2+ permeability and inhibition by selective cationic channel blockers such as Mg2+, Zn2+, and MK-801. In this study, we investigated the structural features that control Ca2+ permeation and channel blockade of the NMDA receptor by in vitro mutagenesis and expression in Xenopus oocytes. We constructed a series of mutations with single amino acid substitutions in the second transmembrane segment of NMDAR1 and examined channel properties of the resultant mutants in combined expression with the NMDAR2A subunit. Substitution of the asparagine with either glutamine or arginine altered both the Ca2+ permeability and the sensitivity to blockades by Mg2+ and MK-801. These mutations also reduced the inhibitory effects of Zn2+ and an antidepressant, desipramine. Based on these results, we concluded that an asparagine ring formed in the central part of the channel-forming second transmembrane segments plays a critical role in determining the Ca2+ permeability and the inhibition of open channel blockers. PMID- 8416948 TI - DNase I and micrococcal nuclease analysis of the tomato proteinase inhibitor I gene in chromatin. AB - The chromatin structure of the wound-inducible proteinase inhibitor I gene was investigated in nuclei from leaves of wounded and unwounded tomato plants. DNase I digestion of intact nuclei revealed that the inhibitor I chromatin structure was highly sensitive to the enzyme compared to the inactive ribosomal chromatin. This sensitivity was independent of wounding. Digestion of tomato nuclei with micrococcal nuclease, which produces a nucleosomal ladder from bulk chromatin in agarose gels, supported these observations. Micrococcal nuclease produced only a faint nucleosomal repeat superimposed on a smear for the coding region of the inhibitor I gene, whereas digestion of inactive ribosomal chromatin produced a well defined nucleosomal ladder. Two DNase I-hypersensitive sites were found in the promoter region of the inhibitor I gene. Both were present before and after wound induction. These two constitutive DNase I-hypersensitive sites may correspond to DNA regulatory regions of the gene. The combined results indicate the existence of an open chromatin conformation in the inhibitor I gene DNA region, present before gene induction by wounding. PMID- 8416949 TI - Adenovirus DNA polymerase is a phosphoprotein. AB - Biological activities of many of the eukaryotic DNA replication proteins are modulated by protein phosphorylation. Investigations of the phosphorylation of adenovirus DNA polymerase (AdPol) have been difficult mainly because of its low level of synthesis in adenovirus-infected HeLa cells. However, when AdPol was overproduced using the recombinant vaccinia virus (RV-AdPol) and the baculovirus expression systems, or by a large scale metabolic labeling of adenovirus 2 infected HeLa cells (native AdPol), in vivo phosphorylation of AdPol could be demonstrated. Phosphoamino acid analysis of [32P]AdPol indicated the presence of phosphoserine independent of the source of AdPol. Comparison of tryptic peptide maps of native AdPol and RV-AdPol revealed that the majority of phosphopeptides were common. Fractionation by high performance liquid chromatography and sequencing of one of the major phosphopeptides revealed serine 67 as a site of phosphorylation. Interestingly, this site is located close to the nuclear localization signal of AdPol and has a consensus substrate recognition sequence for histone H1 (cdc2-related) kinases and mitogen-activated protein kinases. Dephosphorylation of AdPol with calf intestinal alkaline phosphatase resulted in significant decrease in its activity in the in vitro DNA replication initiation assay, suggesting that phosphorylation is important for its biological activity. PMID- 8416950 TI - DNA splicing by an active site mutant of Flp recombinase. Possible catalytic cooperativity between the inactive protein and its DNA substrate. AB - Each strand transfer catalyzed by the Flp recombinase is the composite of two transesterification reactions. The active nucleophilic species in the two reactions are the catalytic site tyrosine (Tyr-343) of Flp and the 5'-hydroxyl from the Flp-nicked DNA substrate, respectively. A "half recombination site" is capable of undergoing this pair of transesterifications in the presence of Flp. When the substrate is a half-site containing a chiral phosphorothioate at the exchange point, the Flp reaction yields a product in which the phosphate chirality is retained. A mutant of Flp that lacks the active site tyrosine, Flp(Y343F), is incapable of mediating strand transfer in a full-recombination site but can execute strand transfer in a half-site. The efficiency of this reaction is about 2% of that of the wild type reaction. The activity of Flp(Y343F) is critically dependent on the length of the half-site spacer. Furthermore, in this reaction, the strand cleavage and strand exchange steps cannot be uncoupled. These results strongly suggest a direct attack by the 5' hydroxyl of the half-site spacer on the phosphodiester at the normal strand transfer point. PMID- 8416951 TI - Characterization of the GArC motif. A novel cis-acting element of the human cardiac myosin heavy chain genes. AB - A positive element between positions -924 and -851 and a negative element between -851 and -762 of the 5'-upstream region of the alpha-myosin heavy chain gene were identified through transient transfection assays in primary cultures of neonatal rat heart cells. Subsequent DNase I protection analysis revealed almost identical footprints at two positions (GAAAAATCT at -904 to -896 and GAAAATCT at -823 to 816). We have designated this sequence the GArC motif (for G,AT-rich,C). Gel mobility shift assays demonstrated the formation of specific complexes with GArC oligomers when either rat heart, rat liver, or HeLa cell nuclear extracts were used. Competition studies with unlabeled GArC oligomers resulted in a loss of binding. Oligomers were also made to the Xenopus cytoskeletal actin serum response element and to a segment of the alpha-MyHC gene (AT-core), each with a similar AT-rich core sequence. No detectable loss of binding resulted from the addition of an excess of either of these unlabeled oligomers. Southwestern blot analysis identified several proteins which interacted with the GArC element, suggesting the presence of a group of related trans-acting factors. Analysis of a sequence in the beta-MyHC gene with the same AT-rich core was negative, suggesting a role for the bases surrounding the protected area in binding. We propose that the GArC motif, together with its associated trans-acting factor(s), provides a novel mechanism of transcriptional control in addition to those previously reported for the cardiac myosin heavy chain genes. PMID- 8416952 TI - Genetic and biochemical studies of a mutant Saccharomyces cerevisiae myristoyl CoA:protein N-myristoyltransferase, nmt72pLeu99-->Pro, that produces temperature sensitive myristic acid auxotrophy. AB - Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase (Nmt1p) is an essential enzyme that transfers myristate from CoA to the amino-terminal glycine residue of at least 12 cellular proteins. Its reaction mechanism is Ordered Bi Bi with myristoyl-CoA binding occurring before binding of nascent polypeptides and release of CoA preceding release of the myristoylprotein product. nmt1-72 is a temperature-sensitive allele, identified by Stone et al. (Stone, D. E., Cole, G. M., Lopes, M. B., Goebl, M., and Reed, S. I. (1991) Genes & Dev. 5, 1969-1981) that causes arrest in the G1 phase of the cell cycle due to reduced acylation of Gpa1p. We have recovered this mutant allele and determined that it contains a single point mutation resulting in a Leu99 (CTA) to Pro (CCA) substitution. Addition of > or = 500 microM myristate but not palmitate to synthetic or rich media rescues the growth arrest caused by nmt1-72 at 37-39 degrees C, consistent with the observation that purified nmt72p has reduced affinity for myristoyl-CoA and that exogenous myristate but not palmitate increases cellular myristoyl-CoA pools. Metabolic labeling studies in S. cerevisiae and co-expression of nmt72p with several protein substrates of Nmt1p in Escherichia coli indicate that the Leu99-->Pro substitution causes a reduction in the acylation of some but not all protein substrates. Since formation of a myristoyl-CoA.Nmt1p complex appears to be required for synthesis/formation of a peptide binding site, these defects in acylation appear to arise either because Leu99 is a component of the enzyme's functionally distinguishable myristoyl-CoA and peptide recognition sites or because Pro99 alters the interaction between myristoyl-CoA and enzyme in a way that precludes formation of a normal peptide binding site. The reduction in affinity for myristoyl-CoA produced by Leu99-->Pro in nmt72p is less than that produced by the Gly451-->Asp mutation in nmt181p, which also produces temperature-sensitive myristic acid auxotrophy. Isogenic, haploid strains containing NMT1, nmt1-72, and nmt1-181 do not manifest any obvious differences in steady state levels of the acyltransferases during growth at permissive temperatures or in the biosynthesis of long chain saturated acyl CoAs. The spectrum of cellular N-myristoylproteins whose level of acylation is affected by nmt1-72 and nmt1-181 is distinct.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8416953 TI - Thyroid hormone receptor-induced bending of specific DNA sequences is modified by an accessory factor. AB - Transcriptional regulation by thyroid and steroid hormone receptors requires their recognition and binding of specific DNA sequences. However, little is known about the mechanisms whereby DNA bound receptors regulate transcription. In the present study, we examined the effects of thyroid hormone receptor (TR) binding on DNA conformation using various TR recognition sites contained within sets of circularly permuted flanking sequences. We show that under conditions where TR binds predominantly as monomer, the conformation of a number of binding sites is changed in a manner consistent with receptor induced bending. Despite similar affinities for receptor binding, not all binding sites tested showed evidence for receptor-induced bending. Notably, the conformation of a sequence from the frog vitellogenin 2 gene, which confers a positive transcriptional response when bound by estrogen receptor (ER), but a negative response when bound by TR, appeared to be unaffected by binding of either TR or ER. The observations suggest that the ability of the receptor to alter DNA architecture is strongly dependent on sequence characteristics other than those required for receptor binding. While both partly purified TR from rat liver and TR translated in vitro were able to induce DNA bending, the bend centers and bend angles produced by these different sources of receptor differed. However, addition of a receptor-depleted fraction from the rat liver TR preparation to in vitro translated receptor stimulated TR binding and appeared to form heterodimers with TR. This resulted in changes in both bend centers and bend angles to resemble more closely those produced by native receptor. Together, these results suggest that receptor-induced DNA bending may be specific to TRs and that the position and degree of bending is further modulated by the formation of heterodimers between TRs and accessory protein(s). PMID- 8416954 TI - Thymosin beta 10 and thymosin beta 4 are both actin monomer sequestering proteins. AB - The beta-thymosins are a family of related peptides. Recently, thymosin beta 4 was identified as a significant actin monomer sequestering protein in cells. To determine if other beta-thymosins also bind actin, and how they may participate in the regulation of actin polymerization, we expressed thymosin beta 4 and its major homolog, thymosin beta 10, in bacteria and characterized their interactions with actin. Equilibrium sedimentation studies showed that thymosin beta 4 behaved as a monomeric protein in solution. Both beta-thymosins bound skeletal muscle actin and inhibited actin polymerization with similar Kd values (between 0.7-1 microM). They were not inhibited by polyphosphoinositides. Kinetic measurements showed that at high ratios of beta-thymosin to actin, beta-thymosin decreased the rate of barbed end filament growth. However, in spite of a close agreement between the kinetic and steady state Kd values, the rate of barbed end filament growth was slightly, but reproducibly, larger than expected, and this deviation was particularly noticeable at lower ratios of beta-thymosin to actin. We conclude that unlike profilin, beta-thymosins are primarily actin monomer sequestering proteins, although some aspects of their interactions with actin are still not completely understood. PMID- 8416955 TI - Regulation of morphology by rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in Swiss 3T3 cells. AB - rho GDP dissociation inhibitor (GDI) is an inhibitory GDP/GTP exchange protein for a group of small GTP-binding proteins including at least rhoA p21, rhoB p21, rac1 p21, rac2 p21, and G25K. Microinjection of rho GDI into Swiss 3T3 cells made the cells round and refractile. This morphological change was accompanied by the disappearance of stress fibers. The rho GDI action was prevented by comicroinjection of rho GDI with the guanosine 5'-(3-O-thio)triphosphate (GTP gamma S)-bound form of rhoA p21, but not with the GTP gamma S-bound form of rhoA p21 lacking the C-terminal three amino acids, which was not post-translationally modified with lipids. The GTP gamma S-bound form of rac1 p21, the same form of G25K, the same form of smg p21B, or Ki-rasval12 p21 was ineffective. Microinjection of the bacterial ADP-ribosyltransferase C3 specific for rho p21 into Swiss 3T3 cells induced the similar changes of morphology and stress fibers. This C3 action was not prevented by comicroinjection of C3 with the GTP gamma S bound form of rhoA p21, but was prevented by comicroinjection with the same form of a rhoA p21 mutant which was not ADP-ribosylated by C3. These results indicate that the rho GDI-rho p21 system regulates cell morphology presumably through the actomyosin system in Swiss 3T3 cells. PMID- 8416956 TI - Inhibition of plasmin, urokinase, tissue plasminogen activator, and C1S by a myxoma virus serine proteinase inhibitor. AB - The myxoma and malignant rabbit fibroma poxviruses are lethal tumorigenic viruses of rabbits whose virulence is modulated by the production of a virus-encoded secreted serine proteinase inhibitor, SERP-1. This viral protein was detected in medium harvested from myxoma and malignant rabbit fibroma virus-infected cells, and its inhibitory profile has been characterized by gel and kinetic analysis. SERP-1 forms complexes with and inhibits the human fibrinolytic enzymes plasmin, urokinase, and two-chain tissue-type plasminogen activator (association rate constants 3.4 x 10(4), 4.3 x 10(4), and 3.6 x 10(4) M-1 s-1 respectively). It is also able to inhibit C1S, the first enzyme in the complement cascade with an association rate constant which was unaffected by the addition of heparin (1.3 x 10(3) M-1 s-1). SERP-1 acts as a substrate for and is cleaved by thrombin, porcine trypsin, human neutrophil elastase, porcine pancreatic elastase, thermolysin, subtilisin, bovine alpha-chymotrypsin, and factor Xa. Incubation with kallikrein and cathepsin G had no effect. The structure of SERP-1 has been modeled on other members of the serpin family which revealed the characteristic serpin architecture apart from the absence of the D-helix. Structural analysis and kinetic assays demonstrate that the absence of this region does not prevent inhibitory activity and furthermore allow the identification of cysteine residues involved in internal and intermolecular disulfide bonding. PMID- 8416957 TI - Metalloregulated expression of the ars operon. AB - The plasmid-borne arsenical resistance (ars) operon encodes an arsenical translocating ATPase and confers resistance to antimonials and arsenicals in Escherichia coli by extrusion of the toxic compounds from the cytosol. The trans acting regulatory ArsR protein was shown to bind to a fragment of DNA containing the ars promoter. Hybrid formation of the ArsR protein with a ArsR-beta-lactamase chimeric protein suggested that the active form of the ArsR repressor is a dimer. From footprinting analysis the binding site was defined as a region of imperfect dyad symmetry just upstream of the -35 site. In vivo the operon was derepressed by oxyions of +III oxidation state of arsenic, antimony, and bismuth, as well as arsenate (As(V)), whereas in vitro ArsR protein-operator interaction was reduced by each of those compounds except arsenate, as determined by gel retardation and DNase I and hydroxyl radical footprinting experiments. This indicates that arsenate is not a true inducer and must be reduced to arsenite in vivo to induce. An operator mutant obtained by deletion of the in vitro ArsR-protected DNA sequence exhibited constitutive ars promoter activity, demonstrating that the binding site is the functional target for the ArsR repressor in vivo. PMID- 8416958 TI - The cDNA of the neutrophil antibiotic Bac5 predicts a pro-sequence homologous to a cysteine proteinase inhibitor that is common to other neutrophil antibiotics. AB - Bac5 is a 5-kDa proline- and arginine-rich antibiotic, stored as inactive precursor (proBac5) in the large granules of bovine neutrophils. A full-length cDNA encoding the precursor form of Bac5 has been cloned. The encoded protein (pre-proBac5) has a calculated mass of 20,031 Da and a pI of 9.21. This comprises a putative signal peptide of 29 amino acid residues and a 101-residue pro sequence that precede the mature antibiotic. The pro-sequence is acidic and may neutralize the highly cationic Bac5, thus accounting for the inactivation of the antibiotic activity observed in in vitro experiments. The structure of mature Bac5 agrees closely with the amino acid sequence previously determined, with an additional tripeptide tail predicting carboxyl-terminal amidation. A valyl residue is deduced at the cleavage site for the proteolytic maturation of proBac5, consistent with a previous observation showing elastase as the enzyme involved in this processing step. The region upstream of Bac5 reveals high identity to corresponding regions of two neutrophil antimicrobial polypeptides, CAP18 from rabbit and bovine indolicidin. The COOH-terminal sequences of these antibiotics are completely unrelated. The proregion also exhibits remarkable similarity to pig cathelin, an inhibitor of cathepsin L, indicating a common evolutionary origin. PMID- 8416959 TI - Expression, purification, and characterization of the yeast KEX1 gene product, a polypeptide precursor processing carboxypeptidase. AB - The Saccharomyces cerevisiae KEX1 gene encodes a protease with carboxypeptidase B like activity involved in K1 and K2 killer toxins and alpha-factor (mating pheromone) precursors processing. The gene has been expressed using the baculovirus/insect cell system, and the KEX1 encoded protein (Kex1p) was purified to apparent homogeneity from detergent-solubilized membrane preparations of insect cells infected with the recombinant virus. The specific activity of the enzyme was enriched 126-fold as compared with the cell lysate, with a recovery of 29%. The NH2-terminal sequence of the purified active enzyme was identical to the predicted sequence after the removal of the signal peptide. This provides evidence that Kex1p, at least in insect cells, is not made as a proenzyme. The optimum pH for activity was 6.0, and the apparent pI value of the protein was below pH 3.0. The enzyme cleaves arginine or lysine from the COOH terminus of synthetic peptides: benzoyl-Phe-Ala-Arg (Km = 284 microM), furylacryloyl (fa)-Ala Arg (Km = 516 microM), and fa-Ala-Lys (Km = 962 microM). The kinetic data obtained reveals that Kex1p preferentially cleaves the COOH-terminal arginine of peptides over the COOH-terminal lysine. Insect-derived Kex1p processes alpha factor-Lys-Arg, its known natural substrate, to mature active alpha-factor, and this maturation event takes place in a sequential manner. Furthermore, the enzyme expresses very high affinity for the 15-amino acid-long peptide, alpha-factor-Lys Arg (Ki = 22 microM), and somewhat lower affinity for the heptapeptides [Leu]enkephalin-Arg-Arg,-Arg-Lys, and [Met]enkephalin-Lys-Lys (Ki = 45, 57, and 81 microM, respectively). The data demonstrate that processing at the COOH terminus of the peptides tested stops after the cleavage of the Arg and/or Lys residues. The specificity of the enzyme for COOH-terminal basic amino acid residues of the peptides used in this study and its high affinity for alpha factor-Lys-Arg confirms the role that Kex1p plays in polypeptide precursor processing in yeast. PMID- 8416960 TI - A cis-acting element accounts for a conserved methylation pattern upstream of the mouse adenine phosphoribosyltransferase gene. AB - A 2.1-kilobase pair region located just upstream of the mouse aprt (adenine phosphoribosyltransferase) gene has a methylation pattern that is conserved in mouse tissues and culture cell lines. This upstream region includes four HpaII/MspI sites. Two of these sites are fully methylated, one is partially methylated, and one is unmethylated. Transfection experiments have demonstrated that the conserved methylation pattern can be reproduced in a mouse embryonal carcinoma stem cell line via de novo methylation (Turker, M.S., Mummaneni, P., and Bishop, P.L. (1991) Somat. Cell Mol. Genet. 17, 151-157). To examine the molecular basis of the conserved methylation pattern, a plasmid-based deletion analysis was conducted by removing and rearranging specific portions of the upstream region. Unmethylated versions of these plasmid constructs were then transfected into the mouse stem cell line and the methylation status of the remaining HpaII/MspI sites determined with a Southern blot analysis. By using this approach, a cis-acting sequence within the upstream region of approximately 0.8 kilobase pairs was identified which appears responsible for the conserved methylation pattern. We use the term "de novo methylation center" to denote this sequence. Based on the results obtained, a model is offered to explain the formation of the conserved methylation pattern in the upstream region. PMID- 8416961 TI - The yeast VPS17 gene encodes a membrane-associated protein required for the sorting of soluble vacuolar hydrolases. AB - vps17 mutants missort and secrete several vacuolar hydrolases. To analyze the role of the VPS17 gene in vacuolar protein delivery, we have cloned this gene by complementation of the vacuolar protein sorting defects of a vps17-5 mutant. Disruption of the VPS17 gene had no effect on the viability of haploid yeast cells, although they show an obvious defect in vacuolar morphology. vps17 disrupted cells contain numerous small vacuole-like compartments and also exhibit a severe defect in the sorting of carboxypeptidase Y (CPY), a soluble vacuolar hydrolase. 95% of CPY is missorted and secreted from the mutant cells. Vacuolar sorting of two other soluble hydrolases, proteinase A and proteinase B, is also affected, but to a lesser extent. Delivery and maturation of the vacuolar membrane protein alkaline phosphatase does not appear to be affected in a delta vps17 strain. The DNA sequence of the VPS17 clone indicates that the gene encodes a 551-amino-acid protein with a calculated molecular mass of 63.1 kDa. The protein sequence is hydrophilic and contains no obvious N-terminal signal sequence or hydrophobic membrane-spanning domains, indicating that the Vps17p does not enter the secretory pathway. Using a Vps17p-specific polyclonal antiserum, we have demonstrated that the Vps17 protein is not modified with N linked carbohydrates at any of its four potential N-linked glycosylation sites. The Vps17 protein, however, fractionates to a particulate fraction after centrifugation at 100,000 x g. Vps17p can be released from this particulate fraction by treatment with either Triton X-100 or urea, indicating that the Vps17p is peripherally associated with a crude membrane fraction. Based on these results, we propose that the Vps17p functions on the cytoplasmic surface of some intracellular organelle, possibly the Golgi complex or an intermediate in Golgi to vacuole transport, to facilitate the sorting and delivery of soluble vacuolar hydrolases. Vacuolar membrane protein traffic, however, appears to occur by a mechanism that is independent of Vps17p function. PMID- 8416962 TI - Effects of the imino sugar N-butyldeoxynojirimycin on the N-glycosylation of recombinant gp120. AB - The imino sugar N-butyldeoxynojirimycin (NB-DNJ) exhibits anti-HIV activity in vitro and inhibits the purified glycoprocessing enzyme alpha 1,2-glucosidase I. It has been speculated that the anti-viral activity of this compound may result from inhibition of HIV envelope glycoprotein processing. However, structural evidence that glucosidase inhibition takes place in intact cells at the anti viral concentration (0.5 mM) is lacking. In this study, N-linked glycosylation of recombinant gp120 expressed in Chinese hamster ovary cells cultured in the presence or absence of NB-DNJ has been characterized. Immunoprecipitation, in conjunction with endoglycosidase H (endo H) digestion and SDS-polyacrylamide gel electrophoresis analysis, revealed that the glycosylation of gp120 was profoundly altered in the presence of NB-DNJ. The majority of the gp120 oligosaccharides from untreated cells were resistant to endo H. However, nearly complete endo H sensitivity was observed following treatment with 0.5 mM NB-DNJ indicating that gp120 expressed in treated cells carries immature, high mannose type oligosaccharides. In addition, using metabolic labeling with [3H]mannose, gel filtration chromatography, and digestion with highly purified glucosidases I and II, we provide the first definitive evidence that glucosidase I inhibition occurs at the anti-viral concentration of NB-DNJ. These data indicate that glucosidase inhibition is a candidate mechanism for the anti-viral activity of this compound. PMID- 8416963 TI - Expression of mammalian 5-aminolevulinate synthase in Escherichia coli. Overproduction, purification, and characterization. AB - 5-Aminolevulinate synthase catalyzes the first step of the heme biosynthetic pathway in nonplant higher eukaryotes. A cDNA encoding for the mouse erythroid 5 aminolevulinate synthase (Schoenhaut, D. S., and Curtis, P.J. (1986) Gene (Amst.) 48, 55-63) has been expressed in Escherichia coli, using the alkaline phosphatase promoter, to a level of 50-60% of the total bacterial protein. Aminolevulinate synthase was overexpressed in an active form and, therefore, was able to rescue hemA mutants, which are unable to grow in the absence of 5-aminolevulinate. A simple purification from the aminolevulinate synthase-overproducing bacterial strain yielded approximately 50 mg of protein, in a high state of purity, per liter of bacterial culture. Moreover, the expressed aminolevulinate synthase could be easily concentrated up to 6-8 mg/ml. Significantly, recombinant aminolevulinate synthase retained physical and catalytic properties identical to those of natural sources. These include the dimeric structure, subunit molecular mass, and pyridoxal 5'-phosphate as an essential cofactor. Removal of the pyridoxal 5'-phosphate led to complete loss of activity. However, the apoenzyme could be readily reconstituted by incubation with 20 microM 5'-pyridoxal phosphate. The Km values are 51 mM for glycine and 55 microM for succinyl-CoA, in the same range of the Km values determined for the nonrecombinant enzyme. This report describes the overexpression of a mammalian 5-aminolevulinate synthase in E. coli and its purification from an overproducing strain. The ready availability of the pure, cloned, sequenced erythroid 5-aminolevulinate synthase makes it possible now for questions pertinent to the enzyme's structure, mechanism, and regulation to be addressed. PMID- 8416964 TI - Cumene hydroperoxide-mediated inactivation of cytochrome P450 2B1. Identification of an active site heme-modified peptide. AB - Cumene hydroperoxide (CuOOH)-mediated inactivation of cytochromes P450 (P450) results in the degradation of their prosthetic heme to products that alkylate the apoprotein. Indirect approaches suggest that this alkylation occurs at the active site. in order to identify the specific apoprotein site(s) alkylated, purified 3H or 14C-heme-labeled P450 2B1 was incubated with CuOOH and subjected to lysyl endopeptidase-C digestion. Two major peaks (L1 and L2) containing 3H- or 14C labeled peptides were detected by reverse-phase high pressure liquid chromatography of the digest. L1 contained the highest specific radioactivity and after Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielded 3 peptide bands (M(r) approximately 3,500 (P1), 5,000 (P2), and 7,000 (P3)). Although all 3 bands were found radiolabeled, the yield of P1 was higher than that of P2 or P3. Amino acid sequence analysis of the first 13 N-terminal residues of P1 revealed the sequence RICLGEGIARNEL, corresponding to residues 434 446 of the reported 2B1 sequence. A species with the molecular mass of 3771 +/- 1 Da was detected in preliminary electrospray mass spectrometric analysis of L1. Since the theoretical average mass of the predicted peptide (residues 434-466) is 3721.99 Da, the additional 49 +/- 1 Da are considered to be contributed by the alkylating heme fragment. This alkylated 2B1 sequence contains not only Cys436, the conserved residue that provides the SH ligand for heme, but also other highly conserved residues, and therefore corresponds to the heme-sandwiching helix L of P450cam. To our knowledge, this is the first report to localize CuOOH-induced heme alkylation of 2B1 to its active site. PMID- 8416965 TI - Prokaryotic elongation factor Tu is phosphorylated in vivo. AB - Covalent modification of proteins by phosphate transfer reactions constitutes a major mechanism of regulation in higher eukaryotes. Recently, phosphorylation of eukaryotic elongation factors has been described. Analysis of Escherichia coli proteins revealed several of them to be phosphorylated. Various lines of evidence lead us to conclude that one of these proteins is identical to elongation factor (EF) Tu, which can be phosphorylated in vivo at one of its threonine residues. Structural analysis showed that one fragment of the phosphorylated EF-Tu is highly resistant to tryptic digestion. Phosphorylation of eubacterial EF-Tu is not restricted to the E. coli factor but could also be demonstrated for Thermus thermophilus HB8 EF-Tu. Overexpression of tufA did not increase the number of EF Tu molecules to be phosphorylated. This may indicate that a constant but limited amount of EF-Tu is modified, possibly for a specific function. Phosphorylation of EF-Tu could also be demonstrated in vitro. Upon analysis of subcellular fractions the highest kinase activity was found in the ribosomal fraction of E. coli. Protein sequencing of both the in vivo and in vitro phosphorylated protein revealed position 382 as the modified threonine residue. PMID- 8416966 TI - Identification of the Alzheimer beta/A4 amyloid precursor protein in clathrin coated vesicles purified from PC12 cells. AB - The Alzheimer beta/A4 amyloid precursor protein (APP) can be proteolytically processed by at least two separate pathways in PC12 cells: chloroquine insensitive secretory cleavage and chloroquine-sensitive intracellular degradation, presumably in the endosomal/lysosomal system. To further investigate the possibility of APP processing in the endosomal/lysosomal system, we have examined whether APP is present in clathrin-coated vesicles (CCVs), which mediate the transport of many proteins to the endosomal compartment. Using a procedure derived from established protocols for the purification of CCVs from mammalian organs, we obtained from PC12 cells highly purified CCVs that displayed the same morphological features as described for CCVs purified from other sources. The CCVs were enriched in full-length mature (fully post-translationally modified) forms of APP, as well as in the carboxyl-terminal APP fragment produced by the secretory cleavage pathway. As CCVs are known to be involved in only two intracellular pathways (trafficking from the plasma membrane to early endosomes, and from the trans-Golgi network to late endosomes/prelysosomes), these findings provide direct evidence that APP is transported to the endosomal/lysosomal system. Furthermore, the presence in CCVs of the carboxyl-terminal fragment resulting from APP secretory cleavage suggests that APP secretory processing occurs in a pre-CCV compartment. PMID- 8416967 TI - The proto-oncogene c-myb mediates an intracellular calcium rise during the late G1 phase of the cell cycle. AB - The intracellular concentration of ionized calcium is involved in regulating mitosis. However, little is known about intracellular levels of calcium during G1. We have demonstrated in vascular smooth muscle cells a mid-G1 decrease in ionized calcium concentration followed by a 2-fold rise at the G1/S interface (44 nM +/- 0.6 nM versus 98 nM +/- 1.1 nM, p < 0.01). The elevation of intracellular calcium is preceded by an increase in c-myb mRNA levels and is abolished with antisense but not missense c-myb oligonucleotides. Furthermore, cells stably transfected with c-myb show a similar 2-fold augmentation in intracellular calcium concentrations, as compared with untransfected cells, which is also abolished by antisense c-myb oligonucleotides. The c-myb-induced rise in intracellular calcium is dependent upon the presence of extracellular calcium and is not suppressed by L type calcium channel blockers. We conclude that c-myb induces an elevation in intracellular calcium levels of vascular smooth muscle cells at the G1/S interface which provides a novel role for this proto-oncogene as well as a potentially important control point for cell cycle regulation. PMID- 8416968 TI - m-Calpain requires DNA for activity on nuclear proteins at low calcium concentrations. AB - m-Calpain (calpain II, m-CANP), which normally requires millimolar Ca2+ for activity in vitro, was capable of proteolyzing a number of matrix proteins in isolated rat liver nuclei at Ca2+ concentrations as low as 3 microM (Mellgren, R. L. (1991) J. Biol. Chem. 266, 13920-13924). Treatment of nuclei with deoxyribonuclease I eliminated the activity of m-calpain at low Ca2+ concentrations, while ribonuclease A and phospholipase C had no effect. Addition of DNA to DNase-treated nuclei restored m-calpain activity at low Ca2+. RNA had little if any effect. Eukaryotic and prokaryotic DNA were equally effective, and synthetic polydeoxyribonucleotides were also activators. m-Calpain did not bind to a DNA-cellulose column in the presence of 200 microM Ca2+, and m-calpain preincubated in the presence of DNA and 200 microM Ca2+ was not activated at low Ca2+ concentrations following removal of the DNA. DNA did not alter the Ca2+ requirement for m-calpain-catalyzed cleavage of casein. These results demonstrate that the Ca2+ requirement for proteolysis of nuclear matrix proteins by m-calpain can be dramatically decreased in the presence of DNA. Activation did not seem to be a result of DNA binding directly to calpain but appeared to require interaction of DNA, calpain, and calpain substrates in the nuclear matrix. PMID- 8416969 TI - S-adenosylhomocysteine hydrolase. Stereochemistry and kinetics of hydrogen transfer. AB - The stereochemistry and kinetics for hydrogen transfer to the catalytically essential NAD+ of S-adenosylhomocysteine hydrolase (SAHase) have been determined for selected adenosine analogues. Reduced SAHase (ENADH), which was made by reconstituting apoSAHase with NADH, stereospecifically transferred the pro-R hydrogen of NADH to 3'-ketoadenosine, a proposed reaction intermediate. Reduction of SAHase by 5'-deoxyadenosine, alpha-L-lyxofuranosyladenine, or adenosine-5' carboxylate quenched the intrinsic protein fluorescence and increased the absorbance of the enzyme at 330 nm. The kinetics for reduction were consistent with a two-step mechanism [formula: see text] where E.L was an initial complex between ligand (L) and enzyme (E), and E'.L' was a complex between reduced enzyme and oxidized ligand. Initial complex formation between SAHase and alpha-L lyxofuranosyladenine (k-1/k-1 = 22.6 microM) was too rapid to be followed by stopped-flow spectrofluorometry at 25 degrees C. The first-order rate constants for reduction (k 2) and oxidation (k-2) of the enzyme were 7.7 and 0.22 s-1, respectively. The dissociation constant for initial complex formation between 5' deoxyadenosine and SAHase was 24 microM; k2 and k-2 were 8.4 x 10(-3) and 4.2 x 10(-3) s-1, respectively. The association rate constant for binding of adenosine 5'-carboxylate to SAHase (k 1) was 2.1 x 10(5) M-1 s-1, and the dissociation rate constant (k-1) was 0.15 s-1; k 2 and k-2 were 0.1 and 0.15 s-1, respectively. The association rate constant for adenine was 5.5 x 10(6) M-1 s-1, and the dissociation rate constant was 6.4 s-1. PMID- 8416970 TI - Subcellular compartmentation of penicillin biosynthesis in Penicillium chrysogenum. The amino acid precursors are derived from the vacuole. AB - The cellular localization of the origin of alpha-aminoadipate used in penicillin biosynthesis and the first enzymic step in Penicillium chrysogenum involved, delta-(alpha-aminoadipyl)-L-cysteinyl-D-valine synthetase (ACVS), has been studied. Subcellular fractions were obtained from protoplasts of a high penicillin-producing strain upon lysis by Triton X-100, and vacuoles purified from them. They were identified by the aid of alpha-mannosidase as a marker enzyme, by the presence of polyphosphate, and their ability to sequester [14C]lysin, added to the protoplasts prior to subcellular fractionation. 15.6 and 26.5%, respectively, of 6-[14C]alpha-aminoadipate, and 8.5 and 10.3%, respectively, of [14C]valine added accordingly were also found in the vacuole, and the higher proportion was found in vacuoles isolated from penicillin producing mycelia. ACVS protein was detected in the membrane as well as the soluble fraction of the purified vacuoles. We propose therefore that ACVS is located either within or bound to the vacuolar membrane, and that the precursor amino acids for penicillin biosynthesis are withdrawn from the vacuolar amino acid pool. PMID- 8416971 TI - Yeast tRNA-splicing endonuclease cleaves precursor tRNA in a random pathway. AB - Introns interrupt many of the tRNA genes of Saccharomyces cerevisiae at a constant position in the anticodon loop. Pre-tRNA transcripts must be accurately cleaved at 3' and 5' splice sites by tRNA endonuclease to release these introns. In order to study splice site cleavage order, substrates were prepared in which the ribose 2'-OH at each of the splice sites was phosphorylated. This modification blocked cleavage by the endonuclease. We found that whichever splice site was blocked the endonuclease can cleave the other site, indicating that the two splice sites were cleaved independently. The endonuclease also cleaved both 3'- and 5'-nicked pre-tRNA(Phe). In addition, both kinds of "2/3 molecules" (exon+intron) were observed in kinetic studies, indicating that they were true biochemical intermediates. The rates of cleavage at the 3' and 5' splice sites of pre-tRNA were compared in several ways. The results showed that the endonuclease cleaves 3' and 5' sites at almost the same rate in the first cleavage, whereas in the second cleavage the 3' site was cleaved faster, indicating that the rates of the two routes for cleavage were unequal. These results demonstrated that the endonuclease cleaved pre-tRNA in a random order, creating two routes for removal of introns from pre-tRNA. PMID- 8416972 TI - Autocrine transforming growth factor-alpha is associated with progression of transformed properties in human colon cancer cells. AB - The GEO colon carcinoma cell line is weakly tumorigenic in athymic mice and shows differentiated properties both in tissue culture and in xenografts. Proliferating monolayer cultures of GEO cells which normally require exogenous epidermal growth factor (EGF) for optimal growth displayed a marked inhibition in growth upon addition of antibodies that block binding to the EGF receptor or neutralize TGF alpha. These results indicated that GEO cells utilize TGF-alpha in a weak autocrine loop. The availability of a weakly malignant model system in which TGF alpha had demonstrable, but low level autocrine activity, permitted the investigation of the role of TGF-alpha in tumorigenesis by generating a stronger autocrine loop through the overexpression of the polypeptide. GEO cells were electroporated with an expression vector containing the human TGF-alpha cDNA, and stable clones were isolated that constitutively expressed the TGF-alpha cDNA in a strong autocrine loop. However, the growth rate of the parental cells in EGF supplemented medium was the same as that of transfected cells with or without growth factor-supplemented medium. Thus, any biological changes generated by the overexpression of TGF-alpha were due to the autocrine nature of the growth mechanism rather than due to any decrease in doubling time leading to a faster growth rate. Transfected GEO cells showed an increase in anchorage-independent growth and formed tumors more readily in athymic nude mice indicating that TGF alpha plays a role in progression of transformed properties. PMID- 8416973 TI - Reticulocalbin, a novel endoplasmic reticulum resident Ca(2+)-binding protein with multiple EF-hand motifs and a carboxyl-terminal HDEL sequence. AB - A novel Ca(2+)-binding protein, tentatively designated reticulocalbin, has been identified and characterized. Reticulocalbin is a luminal protein of the endoplasmic reticulum with an M(r) of 44,000 as revealed by biochemical analysis and immunofluorescence staining. The cDNA of reticulocalbin encodes a protein of 325 amino acids with an amino-terminal signal sequence of 20 amino acids. The protein has six repeats of a domain containing the high affinity Ca(2+)-binding motif, the EF-hand. Although oxygen-containing amino acids important for the positioning of Ca2+ are conserved in all six domains, the conserved glycine residues in the central portion of the EF-hand motif are absent in three of them. Calcium blots showed that recombinant reticulocalbin expressed in bacterial cells binds Ca2+. The protein has the sequence His-Asp-Glu-Leu (HDEL) at its carboxyl terminus. This is similar to the Lys-Asp-Glu-Leu sequence, which serves as a signal to retain the resident proteins in the endoplasmic reticulum of animal cells. A mutant protein lacking the HDEL sequence produced by in vitro mutagenesis has been shown to be secreted into medium in transient expression assays. PMID- 8416974 TI - Identification through overexpression and tagging of the variant type of the mouse H1e and H1c genes. AB - Multiple, nonallelic forms of histone H1 are found in most mammalian tissues. Attempts to determine a functional significance to this diversity is hindered by a paucity of primary amino acid sequence data. Although numerous H1 genes have been cloned, the type of variant they encode cannot be determined by sequence analysis alone. We used transformation and overexpression methods to determine that two cloned mouse H1 genes, MH143 and MH175, encode H1c and H1e, respectively. Since these genes have been completely sequenced, these results establish the amino acid sequence of these variants. Assignment was accomplished by mutagenically "tagging" the genes by incorporation of a codon for methionine, which is not found in the major somatic H1 variants. These genes were placed under the control of the mouse metallothionein I promoter and introduced into 3T3 cells. Products of these mutagenized genes were detected by [35S]methionine labeling and identified by high performance liquid chromatography and sodium dodecyl sulfate-acid polyacrylamide gel electrophoresis. We were also able to induce major alterations in the normal stoichiometry of chromatin-associated H1 variants through overproduction of H1c and H1e. This had little effect on the growth properties of these transformants. PMID- 8416975 TI - Cell-specific translational regulation of S-adenosylmethionine decarboxylase mRNA. Dependence on translation and coding capacity of the cis-acting upstream open reading frame. AB - The mRNA encoding S-adenosylmethionine decarboxylase (AdoMetDC) has a 330 nucleotide 5'-transcript leader containing an open reading frame (uORF) that codes for the hexapeptide MAGDIS. The uORF restricts the intracellular distribution of AdoMetDC mRNA primarily to monosomes in normal T-lymphocytes and in T-cell lines. In contrast, non-lymphoid cells normally carry an average of seven to nine ribosomes per AdoMetDC mRNA molecule (Hill, J.R., and Morris, D. R. (1992) J. Biol. Chem. 267, 21886-21893). Several alterations abolish the negative regulatory effect of the uORF in T-cells. These include removing the site of translational initiation; weakening the context of the translational initiation site; changing the coding capacity of the fourth, fifth, and sixth codons; increasing the length of the uORF at either the 5' or 3' end; or changing the primary order of the codons. In contrast, altering the nucleic acid sequence of the uORF at degenerative positions without changing the amino acid coding capacity did not cause deregulation. The uORF does not regulate translation in the trans-configuration. Our results support a model in which translation of the uORF generates a nascent hexapeptide that interacts with its translating ribosome to suppress translation of AdoMetDC mRNA in a cell-specific manner. Structural features of the carboxyl-terminal 3 amino acids of the putative hexapeptide govern the interaction of the peptide with a component of the translation machinery. PMID- 8416976 TI - Biochemical characterization and molecular genetics of nine mutants of Penicillium chrysogenum impaired in penicillin biosynthesis. AB - Nine mutants of Penicillium chrysogenum (npe1 to npe8 and npe10) impaired in penicillin biosynthesis were screened after nitrosoguanidine mutation. Mutants npe1, npe4, npe5, npe6, npe7, npe8, and npe10 failed to synthesize significant levels of penicillin, whereas strains npe2 and npe3 synthesized about 20% of the penicillin level produced by the parental strain. Mutants npe5 and npe10 did not show alpha-aminoadipylcysteinyl-valine (ACV) synthetase activity in vitro and did not form ACV in vivo. Immunoblotting analysis of the different mutants using antibodies raised against Aspergillus nidulans ACV-synthetase showed that mutants npe5 and npe10 lacked this multienzyme protein, which in the parental strain had a molecular mass of about 420 kDa, and mutants npe2 and npe3 formed reduced level of this protein. All mutants showed normal levels of isopenicillin N synthase, as shown by Western blot analysis and enzyme assays (except npe10 that lacked this enzyme and npe2 and npe3 that formed reduced levels); npe1, npe4, npe6, npe7, npe8, and npe10 lacked isopenicillin N acyltransferase. Southern hybridizations of total DNA of the parental strain and mutants npe5, npe6, npe8, and npe10 with probes internal to the pcbAB, pcbC, and penDE genes showed that mutants npe5, npe6, and npe8 had the same arrangement of the penicillin gene cluster carrying probably point mutations, but mutant npe10 lacked the three penicillin biosynthetic genes, suggesting that it had suffered a deletion of the entire penicillin cluster. Southern hybridization with a pyrG probe as control and fingerprinting analysis of total DNA of npe10 as compared to several P.chrysogenum strains and other Penicillium and Aspergillus species, confirmed that npe10 is a deletion mutant of P. chrysogenum that had lost the penicillin biosynthetic genes. PMID- 8416977 TI - Partial purification and characterization of two distinct protein kinases that differentially phosphorylate the carboxyl-terminal domain of RNA polymerase subunit IIa. AB - RNA polymerase II is a multisubunit enzyme composed of two large subunits of molecular weight in excess of 100,000 and a collection of 8-10 smaller subunits. The largest subunit, designated IIa, contains at its carboxyl terminus a highly repetitive domain consisting of tandem repeats of the consensus sequence Tyr-Ser Pro-Thr-Ser-Pro-Ser. Extensive phosphorylation within this COOH-terminal domain (CTD) gives rise to subunit IIo which has a markedly reduced mobility in SDS polyacrylamide gel electrophoresis (PAGE) relative to subunit IIa. Recent evidence suggests that RNA polymerase IIA, containing an unphosphorylated CTD, is involved in preinitiation complex assembly, whereas RNA polymerase IIO is involved in elongation. Consequently, CTD phosphorylation is thought to occur after RNA polymerase II has bound to the promoter by a protein kinase that stably associates with the preinitiation complex. We present here the partial purification and characterization of two distinct CTD kinases from a HeLa cell transcription extract. These CTD kinases, designated CTDK1 and CTDK2, are fractionated by chromatography on Mono Q. CTDK1 catalyzes the incorporation of approximately 33 pmol of phosphate/pmol of calf thymus RNA polymerase subunit IIa, almost exclusively on serine. CTDK2 catalyzes the incorporation of approximately 50 pmol of phosphate/pmol of calf thymus subunit IIa, predominantly on serine; appreciable phosphate transfer onto threonine is also observed. Phosphorylation by CTDK2, but not CTDK1, results in a complete mobility shift in SDS-PAGE of subunit IIa to the position of IIo. CTDK1 can utilize ATP, dATP, or GTP as phosphate donor, whereas CTDK2 can utilize only ATP or dATP. The apparent Km for ATP is 30 microM for CTDK1 and 60 microM for CTDK2. CTDK1 and CTDK2 also differ in their protein substrate specificity. CTDK1 phosphorylates casein whereas CTDK2 does not. Neither kinase phosphorylates phosvitin or histone H1 to an appreciable extent. CTDK1 and CTDK2 do not appear to be related to cdc2 kinases as determined by their inability to phosphorylate H1 and their failure to react with antibodies directed against the cdc2 kinase. These results establish that a partially fractionated HeLa transcription extract contains two distinct CTD kinases that differ in their nucleotide requirements and in their patterns of CTD phosphorylation. PMID- 8416978 TI - Regulation of purified type I and type II adenylylcyclases by G protein beta gamma subunits. AB - Type I and type II adenylylcyclases have been purified after expression in Sf9 cells, each by application of a two-step purification protocol. The specific activities of the essentially homogeneous enzymes are approximately 7 and 2 mumol.min-1.mg-1, respectively. Each purified enzyme preparation is activated by Gs alpha, but they are regulated in an opposite fashion by G protein beta gamma subunits. Purified beta gamma inhibits Gs alpha-stimulated type I adenylylcyclase directly, while beta gamma activates type II adenylylcyclase and potentiates the Gs alpha-mediated stimulation of the enzyme. This is the first demonstration of the activation of a purified effector molecule by G protein beta gamma subunits. PMID- 8416979 TI - Poly(ADP-ribose) polymerase stimulates DNA polymerase alpha by physical association. AB - The direct effect of the eukaryotic nuclear DNA-binding protein poly(ADP-ribose) polymerase on the activity of DNA polymerase alpha was investigated. Homogenously purified poly(ADP-ribose) polymerase (5 to 10 micrograms/ml) stimulated the activity of immunoaffinity-purified calf or human DNA polymerase alpha by about 6 to 60-fold in a dose-dependent manner. It had no effect on the activities of DNA polymerase beta, DNA polymerase gamma, and primase, indicating that its effect is specific for DNA polymerase alpha. Apparently, poly(ADP-ribosyl)ation of DNA polymerase alpha was not necessary for the stimulation. The stimulatory activity is due to poly(ADP-ribose) polymerase itself since it was immunoprecipitated with a monoclonal antibody directed against poly(ADP-ribose) polymerase. Kinetic analysis showed that, in the presence of poly(ADP-ribose) polymerase, the saturation curve for DNA template primer became sigmoidal; at very low concentrations of DNA, it rather inhibited the reaction in competition with template DNA, while, at higher DNA doses, it greatly stimulated the reaction by increasing the Vmax of the reaction. By the automodification of poly(ADP-ribose) polymerase, however, both the inhibition at low DNA concentration and the stimulation at high DNA doses were largely lost. Furthermore, stimulation by poly(ADP-ribose) polymerase could not be attributed to its DNA-binding function alone since its fragment, containing only the DNA-binding domain, could not exert full stimulatory effect on DNA polymerase, as of the intact enzyme. Poly(ADP ribose) polymerase is co-immunoprecipitated with DNA polymerase alpha, using anti DNA polymerase alpha antibody, clearly showing that poly(ADP-ribose) polymerase may be physically associated with DNA polymerase alpha. In a crude extract of calf thymus, a part of poly(ADP-ribose) polymerase activity existed in a 400-kDa, as well as, a larger 700-kDa complex containing DNA polymerase alpha, suggesting the existence in vivo of a complex of these two enzymes. PMID- 8416980 TI - Functional redundancy in mitotic force generation. PMID- 8416981 TI - Identification of intermediates in the pathway of protein import into chloroplasts and their localization to envelope contact sites. AB - We have used a hybrid precursor protein to study the pathway of protein import into chloroplasts. This hybrid (pS/protA) consists of the precursor to the small subunit of Rubisco (pS) fused to the IgG binding domains of staphylococcal protein A. The pS/protA is efficiently imported into isolated chloroplasts and is processed to its mature form (S/protA). In addition to the mature stromal form, two intermediates in the pathway of pS/protA import were identified at early time points in the import reaction. The first intermediate represents unprocessed pS/protA bound to the outer surface of the chloroplast envelope and is analogous to a previously characterized form of pS that is specifically bound to the chloroplast surface and can be subsequently translocated in the stroma (Cline, K., M. Werner-Washburne, T. H. Lubben, and K. Keegstra. 1985. J. Biol. Chem. 260:3691-3696.) The second intermediate represents a partially translocated form of the precursor that remains associated with the envelope membrane. This form is processed to mature S/protA, but remains susceptible to exogenously added protease in intact chloroplasts. We conclude that the envelope associated S/protA is spanning both the outer and inner chloroplast membranes en route to the stroma. Biochemical and immunochemical localization of the two translocation intermediates indicates that both forms are exposed at the surface of the outer membrane at sites where the outer and inner membrane are closely apposed. These contact zones appear to be organized in a reticular network on the outer envelope. We propose a model for protein import into chloroplasts that has as its central features two distinct protein conducting channels in the outer and inner envelope membranes, each gated open by a distinct subdomain of the pS signal sequence. PMID- 8416982 TI - Golgi-derived vesicles from developing epithelial cells bind actin filaments and possess myosin-I as a cytoplasmically oriented peripheral membrane protein. AB - In the intestinal brush border, the mechanoenzyme myosin-I links the microvillus core actin filaments with the plasma membrane. Previous immunolocalization shows that myosin-I is associated with vesicles in mature enterocytes (Drenckhahn, D., and R. Dermietzel. 1988. J. Cell Biol. 107:1037-1048) suggesting a potential role mediating vesicle motility. We now report that myosin-I is associated with Golgi derived vesicles isolated from cells that are rapidly assembling brush borders in intestinal crypts. Crypt cells were isolated in hyperosmotic buffer, homogenized, and fractionated using differential- and equilibrium-density centrifugation. Fractions containing 50-100-nm vesicles, a similar size to those observed in situ, were identified by EM and were shown to contain myosin-I as demonstrated by immunoblotting and immunolabel negative staining. Galactosyltransferase, a marker enzyme for trans-Golgi membranes was present in these fractions, as was alkaline phosphatase, which is an apical membrane targeted enzyme. Galactosyltransferase was also present in vesicles immuno-purified with antibodies to myosin-I. Villin, a marker for potential contamination from fragmented microvilli, was absent. Myosin-I was found to reside on the vesicle "outer" or cytoplasmic surface for it was accessible to exogenous proteases and intact vesicles could be immunolabeled with myosin-I antibodies in solution. The bound myosin-I could be extracted from the vesicles using NaCl, KI and Na2CO3, suggesting that it is a vesicle peripheral membrane protein. These vesicles were shown to bundle actin filaments in an ATP-dependent manner. These results are consistent with a role for myosin-I as an apically targeted motor for vesicle translocation in epithelial cells. PMID- 8416983 TI - Ezrin contains cytoskeleton and membrane binding domains accounting for its proposed role as a membrane-cytoskeletal linker. AB - Ezrin, a widespread protein present in actin-containing cell-surface structures, is a substrate of some protein tyrosine kinases. Based on its primary and secondary structure similarities with talin and band 4.1 it has been suggested that this protein could play a role in linking the cytoskeleton to the plasma membrane (Gould, K.L., A. Bretscher, F.S. Esch, and T. Hunter. 1989. EMBO (Eur. Mol. Biol. Organ.), J. 8:4133-4142; Turunen, O., R. Winqvist, R. Pakkanen, K.-H. Grzeschik, T. Wahlstrom, and A. Vaheri. 1989. J. Biol. Chem. 264:16727-16732). To test this hypothesis, we transiently expressed the complete human ezrin cDNA, or truncated cDNAs encoding the amino- and carboxy-terminal domains of the protein, in CV-1 cells. Protein epitope tagging was used to unambiguously determine the subcellular distribution of the protein encoded by the transfected cDNA. We show that this protein is concentrated underneath the dorsal plasma membrane in all actin-containing structures and is partially detergent insoluble. The amino terminal domain displays the same localization but is readily extractable by nonionic detergent. The carboxy-terminal domain colocalizes with microvillar actin filaments as well as with stress fibers and remains associated with actin filaments after detergent extraction, and with disorganized actin structures after cytochalasin D treatment. Our results clearly demonstrate that ezrin interacts with membrane-associated components via its amino-terminal domain, and with the cytoskeleton via its carboxy-terminal domain. The amino-terminal domain could include the main determinant that restricts the entire protein to the cortical cytoskeleton in contact with the dorsal plasma membrane and its specialized microdomains such as microvilli, microspikes and lamellipodia. PMID- 8416984 TI - Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. AB - Mitotic spindles of Schizosaccharomyces pombe have been studied by EM, using serial cross sections to reconstruct 12 spindles from cells that were ultrarapidly frozen and fixed by freeze substitution. The resulting distributions of microtubules (MTs) have been analyzed by computer. Short spindles contain two kinds of MTs: continuous ones that run from pole to pole and MTs that originate at one pole and end in the body of the spindle. Among the latter there are three pairs of MT bundles that end on fibrous, darkly staining structures that we interpret as kinetochores. The number of MTs ending at each putative kinetochore ranges from two to four; all kinetochore-associated MTs disappear as the spindle elongates from 3-6 microns. At this and greater spindle lengths, there are no continuous MTs, only polar MTs that interdigitate at the spindle midzone, but the spindle continues to elongate. An analysis of the density of neighboring MTs at the midzone of long spindles shows that their most common spacing is approximately 40 nm, center to center, and that there is a preferred angular separation of 90 degrees. Only hints of such square-packing are found at the midzone of short spindles, and near the poles there is no apparent order at any mitotic stage. Our data suggest that the kinetochore MTs (KMTs) do not interact directly with nonkinetochore MTs, but that interdigitating MTs from the two spindle poles do interact to form a mechanically stable bundle that connects the poles. As the spindle elongates, the number of MTs decreases while the mean length of the MTs that remain increases. We conclude that the chromosomes of S. pombe become attached to the spindle by kinetochore MTs, that these MTs disappear as the chromosomes segregate, that increased separation of daughter nuclei is accompanied by a sliding apart of anti-parallel MTs, and that the mitotic processes of S. pombe are much like those in other eukaryotic cells. PMID- 8416985 TI - Tubulation of Golgi membranes in vivo and in vitro in the absence of brefeldin A. AB - Recent in vivo studies with the fungal metabolite, brefeldin A (BFA), have shown that in the absence of vesicle formation, membranes of the Golgi complex and the trans-Golgi network (TGN) are nevertheless able to extend long tubules which fuse with selected target organelles. We report here that the ability to form tubules (> 7 microns long) could be reproduced in vitro by treatment of isolated, intact Golgi membranes with BFA under certain conditions. Surprisingly, an even more impressive degree of tubulation could be achieved by incubating Golgi stacks with an ATP-reduced cytosolic fraction, without any BFA at all. Similarly, tubulation of Golgi membranes in vivo occurred after treatment of cells with intermediate levels of NaN3 and 2-deoxyglucose. The formation of tubules in vitro, either by BFA treatment or low-ATP cytosol, correlated precisely with a loss of the vesicle associated coat protein beta-COP from Golgi membranes. After removal of BFA or addition of ATP, membrane tubules served as substrates for the rebinding of beta COP and for the formation of vesicles in vitro. These results provide support for the idea that a reciprocal relationship exists between tubulation and vesiculation (Klausner, R. D., J. G. Donaldson, and J. Lippincott-Schwartz. 1992. J. Cell Biol. 116:1071-1080). Moreover, they show that tubulation is an inherent property of Golgi membranes, since it occurs without the aid of microtubules or BFA treatment. Finally the results indicate the presence of cytosolic factors, independent of vesicle-associated coat proteins, that mediate the budding/tubulation of Golgi membranes. PMID- 8416986 TI - Suppression of the bimC4 mitotic spindle defect by deletion of klpA, a gene encoding a KAR3-related kinesin-like protein in Aspergillus nidulans. AB - To investigate the relationship between structure and function of kinesin-like proteins, we have identified by polymerase chain reaction (PCR) a new kinesin like protein in the filamentous fungus Aspergillus nidulans, which we have designated KLPA. DNA sequence analysis showed that the predicted KLPA protein contains a COOH terminal kinesin-like motor domain. Despite the structural similarity of KLPA to the KAR3 and NCD kinesin-like proteins of Saccharomyces cerevisiae and Drosophila melanogaster, which also posses COOH-terminal kinesin like motor domains, there are no significant sequence similarities between the nonmotor or tail portions of these proteins. Nevertheless, expression studies in S. cerevisiae showed that klpA can complement a null mutation in KAR3, indicating that primary amino acid sequence conservation between the tail domains of kinesin like proteins is not necessarily required for conserved function. Chromosomal deletion of the klpA gene exerted no observable mutant phenotype, suggesting that in A. nidulans there are likely to be other proteins functionally redundant with KLPA. Interestingly, the temperature sensitive phenotype of a mutation in another gene, bimC, which encodes a kinesin-like protein involved in mitotic spindle function in A. nidulans, was suppressed by deletion of klpA. We hypothesize that the loss of KLPA function redresses unbalanced forces within the spindle caused by mutation in bimC, and that the KLPA and BIMC kinesin-like proteins may play opposing roles in spindle function. PMID- 8416987 TI - Determinants of the translational mobility of a small solute in cell cytoplasm. AB - The purposes of this study were: (a) to measure the translational mobility of a small solute in cell cytoplasm; (b) to define quantitatively the factors that determine solute translation; and (c) to compare and contrast solute rotation and translation. A small fluorescent probe, 2,7-bis-(2-carboxyethyl)-5-(and 6-) carboxyfluorescein (BCECF), was introduced into the cytoplasm of Swiss 3T3 fibroblasts. BCECF translation was measured by fluorescence recovery after photo bleaching; rotation was measured by Fourier transform polarization microscopy. Diffusion coefficients relative to those in water (D/D0) were determined by comparing mobility in cytoplasm with mobility in standard solutions of known viscosity. At isosmotic cell volume, the relative diffusion coefficients for BCECF translation and rotation in cytoplasm were 0.27 +/- 0.01 (SEM, n = 24, 23 degrees C) and 0.78 +/- 0.03 (n = 4), respectively. As cell volume increased from 0.33 to 2 times isosmotic volume, the relative translational diffusion coefficient increased from 0.047 to 0.32, while the relative rotational diffusion coefficient remained constant. The factors determining BCECF translation were evaluated by comparing rotation and translation in cytoplasm, and in artificial solutions containing dextrans (mobile barriers) and agarose gels (immobile barriers). It was concluded that the hindrance of BCECF translation in cytoplasm could be quantitatively attributed to three independent factors: (a) fluid-phase cytoplasmic viscosity is 28% greater than the viscosity of water (factor 1 = 0.78); (b) 19% of BCECF is transiently bound to intracellular components of low mobility (factor 2 = 0.81); and most importantly, (c) translation of unbound BCECF is hindered 2.5-fold by collisions with cell solids comprising 13% of isosmotic cell volume (factor 3 = 0.40). The product of the 3 factors is 0.25 +/- 0.03, in good agreement with the measured D/D0 of 0.27 +/- 0.01. These results provide the first measurement of the translational mobility of a small solute in cell cytoplasm and define quantitatively the factors that slow solute translation. PMID- 8416988 TI - Induction of neuron-specific tropomyosin mRNAs by nerve growth factor is dependent on morphological differentiation. AB - We have examined the expression of brain-specific tropomyosins during neuronal differentiation. Both TmBr-1 and TmBr-3 were shown to be neuron specific. TmBr-1 and TmBr-3 mRNA levels increased during the most active phase of neurite outgrowth in the developing rat cerebellum. In PC12 cells stimulated by nerve growth factor (NGF) to differentiate to the neuronal phenotype, TmBr-1 and TmBr-3 levels increased with an increasing degree of morphological differentiation. Induction of TmBr-1 and TmBr-3 expression only occurred under conditions where PC12 cells were permitted to extend neurites. NGF was unable to maintain levels of TmBr-1 and TmBr-3 with the loss of neuronal phenotype by resuspension of differentiated PC12 cells. The unique cellular expression and regulation in vivo and in vitro of TmBr-1 and TmBr-3 strongly suggests a critical role of these tropomyosins in neuronal microfilament function. The findings reveal that the induction and maintenance of the neuronal tropomyosins is dependent on morphological differentiation and the maintenance of the neuronal phenotype. PMID- 8416989 TI - A human homologue of the rat metastasis-associated variant of CD44 is expressed in colorectal carcinomas and adenomatous polyps. AB - A recently described splice variant of CD44 expressed in metastasizing cell lines of rat tumors has been shown to confer metastatic potential to a non metastasizing rat pancreatic carcinoma cell line and to non-metastasizing sarcoma cells. Homologues of this variant as well as several other CD44 splice variants are also expressed at the RNA level in human carcinoma cell lines from lung, breast, and colon, and in immortalized keratinocytes. Using antibodies raised against a bacterial fusion protein encoded by variant CD44 sequences, we studied the expression of variant CD44 glycoproteins in normal human tissues and in colorectal neoplasia. Expression of CD44 variant proteins in normal human tissues was readily found on several epithelial tissues including the squamous epithelia of the epidermis, tonsils, and pharynx, and the glandular epithelium of the pancreatic ducts, but was largely absent from other epithelia and from most non epithelial cells and tissues. In human colorectal neoplasia CD44 variant proteins, including homologues of those which confer metastatic ability to rat tumors, were found on all invasive carcinomas and carcinoma metastases. Interestingly, focal expression was also observed in adenomatous polyps, expression being related to areas of dysplasia. The distribution of the CD44 variants in human tissues suggests that they play a role in a few restricted differentiation pathways and that in colorectal tumors one of these pathways has been reactivated. The finding that metastasis-related variants are already expressed at a relatively early stage in colorectal carcinogenesis and tumor progression, i.e., in adenomatous polyps, suggests the existence of a yet unknown selective advantage linked to CD44 variant expression. The continued expression in metastases would be compatible with a role in the metastatic process. PMID- 8416990 TI - TGF beta suppresses casein synthesis in mouse mammary explants and may play a role in controlling milk levels during pregnancy. AB - Mammary explants from 14-15-d-pregnant mice synthesize and secrete milk proteins in culture in response to insulin, hydrocortisone, and prolactin. Here we demonstrate that transforming growth factor beta (TGF beta) treatment suppresses, in a dose dependent and reversible manner, the ability of explants to synthesize and secrete milk caseins. TGF beta does not affect the level of casein mRNA within explants but inhibits casein synthesis posttranscriptionally. We also show increased expression of TGF beta 2 and TGF beta 3 in intact mammary gland as pregnancy progresses, with reduced expression of all three TGF betas at the onset of lactation. These findings suggest that endogenously produced TGF beta may limit the accumulation of milk caseins that are produced in the mammary gland during pregnancy. PMID- 8416991 TI - Unconfined lateral diffusion and an estimate of pericellular matrix viscosity revealed by measuring the mobility of gold-tagged lipids. AB - Nanovid (video-enhanced) microscopy was used to determine whether lateral diffusion in the plasma membrane of colloidal gold-tagged lipid molecules is confined or is unrestricted. Confinement could be produced by domains within the plane of the plasma membrane or by filamentous barriers within the pericellular matrix. Fluorescein-phosphatidylethanolamine (F1-PE), incorporated into the plasma membranes of cultured fibroblasts, epithelial cells and keratocytes, was labeled with 30-nm colloidal gold conjugated to anti-fluorescein (anti-F1). The trajectories of the gold-labeled lipids were used to compute diffusion coefficients (DG) and to test for restricted motion. On the cell lamella, the gold-labeled lipids diffused freely in the plasma membrane. Since the gold must move through the pericellular matrix as the attached lipid diffuses in the plasma membrane, this result suggests that any extensive filamentous barriers in the pericellular matrix are at least 40 nm from the plasma membrane surface. The average diffusion coefficients ranged from 1.1 to 1.7 x 10(-9) cm2/s. These values were lower than the average diffusion coefficients (DF) (5.4 to 9.5 x 10( 9) cm2/s) obtained by FRAP. The lower DG is partially due to the pericellular matrix as demonstrated by the result that heparinase treatment of keratocytes significantly increased DG to 2.8 x 10(-9) cm2/s, but did not affect DF. Pericellular matrix viscosity was estimated from the frictional coefficients computed from DG and DF and ranged from 0.5 to 0.9 poise for untreated cells. Heparinase treatment of keratocytes decreased the apparent viscosity to approximately 0.1 poise. To evaluate the presence of domains or barriers, the trajectories and corresponding mean square displacement (MSD) plots of gold labeled lipids were compared to the trajectories and MSD plots resulting from computer simulations of random walks within corrals. Based on these comparisons, we conclude that, if there are domains limiting the diffusion of F1-PE, most are larger than 5 microns in diameter. PMID- 8416992 TI - Extracellular matrix regulates expression of the TGF-beta 1 gene. AB - Transforming growth factor-beta (TGF-beta) is a potent regulator of cell proliferation and modulates the interactions of cells with their extracellular matrix (ECM), in part by inducing the synthesis of various ECM proteins. Three different isoforms of TGF-beta are synthesized in a defined pattern in specific cell populations in vivo. In the specific case of TGF-beta 1, this well-defined and limited expression stands in sharp contrast to its synthesis by virtually all cells in culture. Using mammary epithelial cells as a model system, we evaluated the substratum dependence of the expression of TGF-beta 1. The level of TGF-beta 1 expression is high in cells on plastic, but is strongly downregulated when cells are cultured on a reconstituted basement membrane matrix. In contrast, TGF beta 2 mRNA levels in cells on either substratum remain unchanged. Using the chloramphenicol acetyl transferase gene as reporter gene under the control of the TGF-beta 1 promoter, we show that transcription from this promoter is suppressed when the cells are in contact with either endogenously synthesized or exogenously administered basement membrane. TGF-beta 1 promoter activity is strongly induced by the absence of basement membrane, i.e., by direct contact of the cells with plastic. This modulation of transcription from the TGF-beta 1 promoter occurs in the absence of lactogenic hormones which allow full differentiation. Our results thus indicate that basement membrane is an important regulator of TGF-beta 1 synthesis, and explain why most cells in culture on plastic express TGF-beta 1 in contrast with the more restricted TGF-beta 1 synthesis in vivo. We propose that there is a feedback loop whereby TGF-beta 1-induced synthesis of basement membrane components is repressed once a functional basement membrane is present. Finally, these results together with our current knowledge of regulation of TGF beta 1 and TGF-beta 2 synthesis, suggest that, in vivo, TGF-beta 1 may play a major role in regulating the ECM synthesis and the cell-ECM interactions, whereas TGF-beta 2 may be more important in morphogenetic processes. PMID- 8416993 TI - Integrins in point contacts mediate cell spreading: factors that regulate integrin accumulation in point contacts vs. focal contacts. AB - We have studied the function and distribution of the alpha 1 beta 1, alpha 5 beta 1 and alpha 6 beta 1 heterodimers on type-1 astrocytes with antibodies specific for integrin subunits (alpha 1, alpha 5, alpha 6, and beta 1). The alpha 1 beta 1 heterodimer mediates adhesion to laminin and collagen, the alpha 5 beta 1 to fibronectin in an RGD-dependent manner. The alpha 5 beta 1 integrin is found in focal contacts in long-term cultures of well-spread astrocytes colocalizing with vinculin and the termini of actin stress fibers. alpha 1 beta 1 heterodimers can occasionally be found as small aggregates within focal contacts but they do not accumulate there. Instead, alpha 1 beta 1 integrins are found in punctate deposits called point contacts which are distributed over the upper and the lower cell surfaces whether laminin, collagen, fibronectin or polylysine is used as a substratum. Unlike focal contacts, point contacts contain clathrin but rarely codistribute with actin or vinculin. Two observations indicate that these point contacts are functional. First, mAb 3A3, directed against the rat alpha 1 subunit, inhibits the attachment of astrocytes to laminin and collagen. Second, during the spreading of astrocytes, a band of point contacts forms around the cell perimeter at a time when no focal contacts are visible. While alpha 1 beta 1 integrins are found only in point contacts in astrocytes, the alpha 6 beta 1 integrin, another laminin receptor, is localized within focal contacts. Moreover, alpha 1 beta 1 heterodimers accumulate in focal contacts in fibroblasts. Thus, the alpha subunit contributes, independent of its ligand, to functional integrin heterodimer accumulation in focal contacts or in point contacts. This accumulation varies among different cell types with apparently identical heterodimers as well as with the motile state (spreading vs. flattened) of the same cells. PMID- 8416994 TI - Multiple GTP-binding proteins participate in clathrin-coated vesicle-mediated endocytosis. AB - We have examined the effects of various agonists and antagonists of GTP-binding proteins on receptor-mediated endocytosis in vitro. Stage-specific assays which distinguish coated pit assembly, invagination, and coat vesicle budding have been used to demonstrate requirements for GTP-binding protein(s) in each of these events. Coated pit invagination and coated vesicle budding are both stimulated by addition of GTP and inhibited by GDP beta S. Although coated pit invagination is resistant to GTP gamma S, A1F4-, and mastoparan, late events involved in coated vesicle budding are inhibited by these antagonists of G protein function. Earlier events involved in coated pit assembly are also inhibited by GTP gamma S, A1F4-, and mastoparan. These results demonstrate that multiple GTP-binding proteins, including heterotrimeric G proteins, participate at discrete stages in receptor mediated endocytosis via clathrin-coated pits. PMID- 8416995 TI - Overlapping distribution of two glycosyltransferases in the Golgi apparatus of HeLa cells. AB - Thin, frozen sections of a HeLa cell line were double labeled with specific antibodies to localize the trans-Golgi enzyme, beta 1,4 galactosyltransferase (GalT) and the medial enzyme, N-acetylglucosaminyltransferase I (NAGT I). The latter was detected by generating a HeLa cell line stably expressing a myc-tagged version of the endogenous protein. GalT was found in the trans-cisterna and trans Golgi network but, contrary to expectation, NAGT I was found both in the medial- and trans-cisternae, overlapping the distribution of GalT. About one third of the NAGT I and half of the GalT were found in the shared, trans-cisterna. These data show that the differences between cisternae are determined not by different sets of enzymes but by different mixtures. PMID- 8416996 TI - Phosphorylation of the cation-independent mannose 6-phosphate receptor is closely associated with its exit from the trans-Golgi network. AB - We have previously shown that two serine residues present in two conserved regions of the bovine cation-independent mannose 6-phosphate receptor (CI-MPR) cytoplasmic domain are phosphorylated in vivo (residues 2421 and 2492 of the full length bovine CI-MPR precursor). In this study, we have used CHO cells to investigate the phosphorylation state of these two serines along the different steps of the CI-MPR exocytic and endocytic recycling pathways. Transport and phosphorylation of the CI-MPR in the biosynthetic pathway were examined using deoxymannojirimycin (dMM), a specific inhibitor of the cis-Golgi processing enzyme alpha-mannosidase I which leads to the accumulation of N-linked high mannose oligosaccharides on glycoproteins. Upon removal of dMM, normal processing to complex-type oligosaccharides (galactosylation and then sialylation) occurs on the newly synthesized glycoproteins, including the CI-MPR which could then be purified and analyzed on lectin affinity columns. Phosphorylation of the newly synthesized CI-MPR was concomitant with the sialylation of its oligosaccharides and appeared as a major albeit transient modification. Phosphorylation of the cell surface CI-MPR was examined during its endocytosis as well as its return to the Golgi using antibody tagging and exogalactosylation. The cell surface CI-MPR was not phosphorylated when it entered clathrin-coated pits or when it moved to the early and late endosomes. In contrast, the surface CI-MPR was phosphorylated when it had been resialylated upon its return to the trans-Golgi network. Subcellular fractionation experiments showed that the phosphorylated CI-MPR and the corresponding kinase were found in clathrin-coated vesicles. Collectively, these results indicate that phosphorylation of the two serines in the CI-MPR cytoplasmic domain is associated with a single step of transport of its recycling pathways and occurs when this receptor is in the trans-Golgi network and/or has left this compartment via clathrin-coated vesicles. PMID- 8416997 TI - Consumption of EGF by A431 cells: evidence for receptor recycling. AB - We examined the extent of EGF consumption by EGFR in A431 cells. When 125I-EGF was added to A431 cell cultures at low or high density, at concentrations which corresponded to 10-fold excess of ligand over receptor on the cell surface, most of the 125I-EGF was consumed within 2 h. The amounts of 125I-EGF consumed were much greater than available EGFR on the A431 cells, by a factor of 6.5 in low density cultures and 5.8 in high-density cultures. When the concentration of 125I EGF was increased in low density cultures, further consumption of 125I-EGF by the A431 cells was greatly reduced, partially due to a rapid down regulation of EGFR. However, when higher concentrations of 125I-EGF were added to high density cultures, with reduced receptor down regulation, the cells continued to consume a large fraction of the EGF in the culture medium. The consumption of 125I-EGF by these cultures was in excellent agreement with the measured amount of ligand internalized into the cell. EGF consumption was far in excess of the number of EGFR down regulated or degraded. Only a minor portion of the EGFR could have been replaced during the assay period by synthesis of new EGFR or from the intracellular pool of EGFR, and the fluid-phase uptake of EGF is only temporarily increased by exposure to EGF. Our results demonstrate that EGFR in high density A431 cell cultures recycled many times. The apparent level of recycling was dependent upon the concentration of EGF and followed Michaelis-Menton kinetics for ligand concentrations as high as 215 nM. At this EGF concentration, high density cultures consumed 45 EGF molecules per receptor over a period of 6 h. PMID- 8416998 TI - Reconstitution of protein translocation from solubilized yeast membranes reveals topologically distinct roles for BiP and cytosolic Hsc70. AB - We reconstituted prepro-alpha-factor translocation and signal peptide processing using a yeast microsomal detergent soluble fraction formed into vesicles with soybean phospholipids. Reconstituted translocation required ATP, and was deficient when sec63 and kar2 (BiP) mutant cells were used as a source of membranes. Normal translocation was observed with vesicles reconstituted from a mixture of pure wild-type yeast BiP and a soluble fraction of kar2 mutant membranes. Two other heat-shock cognate (hsc) 70 homologs, yeast cytosolic hsc70 (Ssalp) and E. coli dnaK protein did not replace BiP. Conversely, BiP was not active under conditions where translocation into native ER vesicles required cytosolic hsc70. We conclude that cytosolic hsc70 and BiP serve noninterchangeable roles in polypeptide translocation, possibly because distinct, asymmetrically oriented membrane proteins are required to recruit each protein to opposing surfaces of the ER membrane. PMID- 8416999 TI - Induction of 70-kDa heat shock protein and hsp70 mRNA following transient focal cerebral ischemia in the rat. AB - Induction of the 70-kDa heat shock protein (HSP70) was demonstrated immunocytochemically in adult rats 4 h to 7 days following temporary middle cerebral artery (MCA) occlusions lasting 30, 60, or 90 min. Maximal HSP70 induction occurred approximately 24 h following ischemia. Thirty minutes of ischemia induced HSP70 in neurons throughout the cortex in the MCA distribution, whereas 90 min of ischemia induced HSP70 in neurons in the penumbra. HSP70 protein was induced in endothelial cells in infarcted neocortex following 60-90 min of MCA occlusion, and HSP70 was induced in endothelial cells in infarcted regions of lateral striatum following 30-90 min of MCA occlusion. hsp70 mRNA was induced in the MCA distribution in cortex and to a lesser extent in striatum at 2 h to 3 days following 60 min of ischemia. It is proposed that brief ischemia induces hsp70 mRNA and HSP70 protein in the cells most vulnerable to ischemia- the neurons. HSP70 protein is not induced in most neurons and glia following 60 90 min of ischemia in areas destined to infarct, whereas it is induced in vascular endothelial cells. PMID- 8417000 TI - Immunolocalization of heat shock protein after fluid percussive brain injury and relationship to breakdown of the blood-brain barrier. AB - We have previously developed a model of mild, lateral fluid percussive head injury in the rat and demonstrated that although this injury produced minimal hemorrhage, breakdown of the blood-brain barrier was a prominent feature. The relationship between posttraumatic blood-brain barrier disruption and cellular injury is unclear. In the present study we examined the distribution and time course of expression of the stress protein HSP72 after brain injury and compared these findings with the known pattern of breakdown of the blood-brain barrier after a similar injury. Rats were subjected to a lateral fluid percussive brain injury (4.8-5.2 atm, 20 ms) and killed at 1, 3, and 6 h and 1, 3, and 7 days after injury. HSP72-like immunoreactivity was evaluated in sections of brain at the light-microscopic level. The earliest expression of HSP72 occurred at 3 h postinjury and was restricted to neurons and glia in the cortex surrounding a necrotic area at the impact site. By 6 h, light immunostaining was also noted in the pia-arachnoid adjacent to the impact site and in certain blood vessels that coursed through the area of necrosis. Maximal immunostaining was observed by 24 h postinjury, and was primarily associated with the cortex immediately adjacent to the region of necrosis at the impact site. This region consisted of darkly immunostained neurons, glia, and blood vessels. Immunostaining within the region of necrosis was restricted to blood vessels. HSP72-like immunoreactivity was also noted in a limited number of neurons and glia in other brain regions, including the parasagittal cortex, deep cortical layer VI, and CA3 in the posterior hippocampus.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417001 TI - Hydrogen peroxide production by monoamine oxidase during ischemia-reperfusion in the rat brain. AB - Monoamine oxidase (MAO) as a source of hydrogen peroxide (H2O2) was evaluated during ischemia-reperfusion in vivo in the rat brain. H2O2 production was assessed with and without inhibition of MAO during and after 15 min of ischemia. Metabolism of H2O2 by catalase during ischemia and reperfusion was measured in forebrain homogenates using aminotriazole (ATZ), an irreversible H2O2-dependent inhibitor of catalase. Catecholamine and glutathione concentrations in forebrain were measured with and without MAO inhibitors. During ischemia, forebrain blood flow was reduced to 8% of baseline and H2O2 production decreased as measured at the microperoxisome. During reperfusion, a rapid increase in H2O2 generation occurred within 5 min as measured by a threefold increase in oxidized glutathione (GSSG). The H2O2-dependent rates of ATZ inactivation of catalase between control and ischemia-reperfusion were similar, indicating that H2O2 was more available to glutathione peroxidase than to catalase in this model. MAO inhibitors eliminated the biochemical indications of increased H2O2 production and increased the catecholamine concentrations. Mortality was 67% at 48 h after ischemia reperfusion, and there was no improvement in survival after inhibition of MAO. We conclude that MAO is an important source of H2O2 generation early in brain reperfusion, but inhibition of the enzyme does not improve survival in this model despite ablating H2O2 production. PMID- 8417002 TI - Induction of copper-zinc superoxide dismutase in gerbil hippocampus after ischemia. AB - To assess the role of Cu-Zn superoxide dismutase (CuZnSOD) in regulating cellular antioxidant defenses, we studied the induction of CuZnSOD mRNA by an in situ hybridization technique and of CuZnSOD protein by an immunocytochemical method in the gerbil hippocampus following 5 min of transient global ischemia. For hybridization, we synthesized 48-mer oligonucleotide (base 465-512) complementary to rat CuZnSOD mRNA. Northern blot analysis showed hybridization to a single band of molecular weight 0.65 kb. After 5 min of ischemia, the signal became stronger at 3 and 24 h and returned to the control level 3 days later. In situ hybridization histochemistry revealed an increase in labeling throughout the hippocampus, especially in the granular layer 3 h following ischemia. The increase was prolonged only in the CA1 pyramidal layer after 24 h and was eliminated within 3 days or later. Conversely, analysis by Western blotting revealed that the insult produced few effects on the induction of CuZnSOD protein. Immunocytochemistry for CuZnSOD revealed a reduced immunostaining in the CA1 pyramidal layer at 24 h of recirculation when the persistent expression of CuZnSOD mRNA was shown in the same area. Our findings suggest that the expression of endogenous CuZnSOD is temporarily stimulated by an ischemic insult without increasing the protein level. The prolonged increase in mRNA and the decrease in the protein of CuZnSOD in the CA1 neurons seem to imply an important role of the endogenous antioxidant enzyme that protects against the detrimental effects of superoxide radicals on delayed neuronal death. PMID- 8417003 TI - Spectral analysis of dynamic PET studies. AB - We describe a new technique for the analysis of dynamic positron emission tomography (PET) studies in humans, where data consist of the time courses of label in tissue regions of interest and in arterial blood, following the administration of radiolabelled tracers. The technique produces a simple spectrum of the kinetic components which relate the tissue's response to the blood activity curve. From this summary of the kinetic components, the tissue's unit impulse response can be derived. The convolution of the arterial input function with the derived unit impulse response function gives the curve of best fit to the observed tissue data. The analysis makes no a priori assumptions regarding the number of compartments or components required to describe the time course of label in the tissue. Rather, it is based on a general linear model, presented here in a formulation compatible with its solution using standard computer algorithms. Its application is illustrated with reference to cerebral blood flow, glucose utilization, and ligand binding. The interpretation of the spectra, and of the tissue unit impulse response functions, are discussed in terms of vascular components, unidirectional clearance of tracer by the tissue, and reversible and irreversible phenomena. The significance of the number of components which can be identified within a given datum set is also discussed. The technique facilitates the interpretation of dynamic PET data and simplifies comparisons between regions and between subjects. PMID- 8417004 TI - Creatine kinase-catalyzed reaction rate in the cyanide-poisoned mouse brain. AB - Brain creatine kinase (CK)-catalyzed phosphorus flux from phosphocreatine (PC) to ATP was measured in vivo in young adult mice made reversibly hypoxic by injection of cyanide. Phosphorus spectra and saturation transfer measurements of CK catalyzed flux were acquired using a high-field (8.45 T) nuclear magnetic resonance (NMR) spectrometer. After low cyanide doses (1-3 mg/kg of body weight), there were no measurable changes in brain pH or in concentrations of PC, the nucleoside triphosphates (including ATP), and Pi. The CK-catalyzed phosphorus flux increased about 75% after the low cyanide dose. Higher doses (4-6 mg/kg) produced a transient 30-40% decrease in PC concentration, doubling of Pi, and a 0.2 unit decrease in pH. The CK-catalyzed phosphorus flux decreased 50-80% after the higher cyanide doses. This decrease in phosphorus flux was present long after reactant concentrations returned to precyanide values. It is proposed that the increase in brain CK-catalyzed phosphorus flux with the lower cyanide doses is due to an increase in ADP concentration. The large, prolonged decrease in CK catalyzed reaction rate in the moderately poisoned brain may be due to loss of activity of the mitochondrial CK isoform. PMID- 8417005 TI - Glial glycogen stores affect neuronal survival during glucose deprivation in vitro. AB - Glia perform several energy-dependent functions that may aid neuronal survival under pathological conditions. Glycogen is the major energy reserve in brain, and it is localized almost exclusively to astrocytes. Using murine cortical cell cultures containing both glia and neurons, we examined the effect of altered glial glycogen stores on neuronal survival following glucose deprivation. As previously reported, cultures exposed for several hours to media lacking glucose developed widespread neuronal degeneration without glial degeneration. If glial astrocyte glycogen content was increased to 2-3 times control levels by a 24-h pretreatment with 1 microM insulin or 0.5 mM methionine sulfoximine (MSO), glucose deprivation-induced neuronal degeneration was attenuated. These protective effects were blocked if glycogen levels were reduced back to control levels by a 30-min exposure to 1 mM dibutyryl cyclic AMP or 20 microM norepinephrine prior to glucose deprivation. Astrocyte glycogen stores may be an important factor influencing neuronal survival under conditions of energy substrate limitation. PMID- 8417006 TI - Selective vulnerability of white matter during spinal cord ischemia. AB - The long-term effects of spinal cord ischemia were studied in 21 rats by lesion scores (LS, n = 21), somatosensory evoked potentials (SEP, n = 16), electromyographic measurements (EMG, n = 12) and histology of the spinal cord (n = 21) 48.5 +/- 57.2 days after 10- to 12-min occlusion of the thoracic aorta and subclavian arteries. All the animals were initially paraplegic with a spastic presentation but seven recovered within 2 days (group A), demonstrating low LS (3.4 +/- 1.05) normal EMGs (n = 3) and unremarkable histology. The 14 paraplegic animals presented relevant findings of the lumbar cord consisting of white matter lesions only (group B, n = 7) or white and gray matter lesions (group C, n = 7). Group B animals showed severe deficit (LS = 11.8 +/- 2.93) without denervation on EMG (n = 5) or muscle atrophy on histology. Group C animals displayed equal impairment (LS = 14.4 +/- 0.71), denervation on EMG (n = 4), and muscle atrophy. Resting motor unit activity of groups B and C were significantly different from group A (p < 0.001), while LS of groups B and C did not differ (p = 0.083). These data underscore the nature and the extent of white matter lesions during spinal cord ischemia, a finding which has generally been eclipsed by emphasis on gray matter lesions in previous studies. PMID- 8417007 TI - Influence of plasma glucose concentration on rat brain extracellular calcium transients during spreading depression. AB - The objective of this study was to establish whether tissues that are energy compromised, but not energy depleted, demonstrate exaggerated calcium transients when subjected to membrane depolarizations of the spreading depression (SD) type. Anesthetized and artificially ventilated rats were given insulin in order to induce progressively lower plasma glucose concentrations. Spreading depression was elicited by local application of KCl; extracellular calcium concentration (Ca2+e) as well as direct current (DC) potential were recorded. When plasma glucose concentration fell below approximately 3 mM, the duration of the Ca2+e transient gradually increased to values exceeding 500% of control. The increase was associated with a corresponding increase in the duration of the DC potential shift, but the amplitude of the Ca2+e transient did not change. It is concluded that a restriction of glucose (or oxygen) supply, as occurs in hypoglycemia (or hypoxia), prolongs the calcium transient associated with depolarization of the SD type, even though tissue phosphocreatinine and ATP concentrations are normal. The results support the contention that repeated depolarizations, occurring in the penumbral zone of a focal ischemic lesion, could lead to calcium-related damage. PMID- 8417008 TI - In memoriam Christian Crone, M.D., Ph.D. (1926-1990). PMID- 8417009 TI - Human striatal L-dopa decarboxylase activity estimated in vivo using 6 [18F]fluoro-dopa and positron emission tomography: error analysis and application to normal subjects. AB - DOPA decarboxylase is the enzyme directly responsible for the synthesis of the neurotransmitters dopamine and serotonin, and indirectly of noradrenaline, in brain. We used the decarboxylation coefficient (k3D) of 6-[18F]fluoro-DOPA (FDOPA) to denote the relative activity of L-DOPA decarboxylase in vivo in the human brain. To determine the relative enzyme activity with positron emission tomography (PET), we evaluated the model that separates the metabolism into compartments of nondiffusible and diffusible (i.e., transient) tracer metabolites. Error analysis indicated that the least-squares optimization alone was not sufficient to yield accurate estimates of k3D in the presence of the inherent error of PET. To improve the accuracy of the k3D estimates by optimizing the number of parameters, we introduced biological constraints which included a tracer partition volume (Ve) common to frontal cortex and striatum, and a fixed ratio (q) between the blood-brain barrier transport coefficients of O-methyl [18F]fluoro-DOPA and FDOPA, the two sources of radioactivity in plasma. We found that a two-step analysis yielded sufficiently accurate estimates of k3D. The two steps include the initial estimation of the partition volume in frontal cortex and the subsequent use of this value to determine k3D in striatum and other structures. We studied twelve healthy controls (age 45 +/- 15 years). The average k3D value was 0.081 +/- 0.024 min-1 (coefficient of variation (COV) 30%) for caudate nucleus, 0.074 +/- 0.013 min-1 (COV 18%) for putamen, and 0.010 +/- 0.005 min-1 (COV 50%) for cerebral cortex. PMID- 8417010 TI - Functional connectivity: the principal-component analysis of large (PET) data sets. AB - The distributed brain systems associated with performance of a verbal fluency task were identified in a nondirected correlational analysis of neurophysiological data obtained with positron tomography. This analysis used a recursive principal-component analysis developed specifically for large data sets. This analysis is interpreted in terms of functional connectivity, defined as the temporal correlation of a neurophysiological index measured in different brain areas. The results suggest that the variance in neurophysiological measurements, introduced experimentally, was accounted for by two independent principal components. The first, and considerably larger, highlighted an intentional brain system seen in previous studies of verbal fluency. The second identified a distributed brain system including the anterior cingulate and Wernicke's area that reflected monotonic time effects. We propose that this system has an attentional bias. PMID- 8417011 TI - 6-[18F]fluoro-L-dopa metabolism in living human brain: a comparison of six analytical methods. AB - In 11 normal volunteers and six patients with Parkinson's disease, we compared six different analyses of dopaminergic function with L-3,4-dihydroxy-6 [18F]fluorophenylalanine (FDOPA) and positron emission tomography (PET). The caudate nucleus, putamen, and several reference regions were identified in PET images, using magnetic resonance imaging (MRI). The six analyses included two direct determinations of DOPA decarboxylase activity (k3D, k3*), the slope intercept plot based on plasma concentration (K), two slope-intercept plots based on tissue content (k3r, k3s), and the striato-occipital ratio [R(T)]. For all analyses, the difference between two groups of subjects (normal volunteers and patients with Parkinson's disease) was larger in the putamen than in the caudate. For the caudate nucleus, the DOPA decarboxylase activity (k3D, k3*), tissue slope intercept plots (kr3, ks3); and striato-occipital ratio [R(T)] analyses significantly discriminated between the normal volunteers and the patients with Parkinson's disease (p < 0.005) [with least significance for k3* (p < 0.05)], while the plasma slope-intercept plot (K) failed to do so. For the putamen, the values for k3D, k3*, K, k3r, k3s, and R(T) of normal volunteers were significantly higher than those of patients (p < 0.005) [with least significance for K (p < 0.025)]. Linear correlations were significant between k3D and k3s; k3D and k3r; k3D and R(T); and k3D and k3*, in this order of significance. We found no correlation between k3D and K values in the caudate nucleus. PMID- 8417012 TI - Nitric oxide synthesis and regional cerebral blood flow responses to hypercapnia and hypoxia in the rat. AB - The role of nitric oxide (NO) synthesis in the cerebral hyperemic responses to hypercapnia and hypoxia was investigated in anesthetized rats. Regional CBF (rCBF) measurements were obtained in the cortex (CX), subcortex (SC), brainstem (BS), and cerebellum (CE) using radiolabeled microspheres. The rCBF responses to either hypercapnia (PaCO2 = 70-80 mm Hg) or hypoxia (PaO2 = 40-45 mm Hg) were compared in rat groups studied in the presence and absence of NO synthase inhibition induced via the intravenous infusion of nitro-L-arginine methyl ester (L-NAME, 3 mg kg-1 min-1). Administration of L-NAME under normocapnic/normoxic conditions produced a 40-60% reduction in baseline rCBF values, indicating the presence of a NO "tone" in the cerebral vasculature. Infusion of L-NAME resulted in a substantial attenuation, in all regions measured, of the rCBF increases that normally accompany hypercapnia. In comparing saline-infused to L-NAME-infused rats, the percentage increases in rCBF (from normocapnic baseline values) were 351% versus 166% (CX), 446% versus 199% (SC), 443% versus 206% (BS), and 483% versus 174% (CE), respectively. The rCBF changes from baseline (delta rCBF in ml 100 g-1 min-1) were 488 versus 57 (CX), 570 versus 60 (SC), 434 versus 72 (BS), and 393 versus 45 (CE), respectively. These differences were all statistically significant (p < 0.05). During hypoxia, when compared to rats not given L-NAME, inhibition of NO synthase activity resulted in significantly greater (p < 0.05) percentage increases in rCBF (from normoxic baseline values) in most regions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417013 TI - Selective reductions in the activity of the pyruvate dehydrogenase complex in mitochondria isolated from brain subregions following forebrain ischemia in rats. AB - Previous studies showed that in rats exposed to 30 min of forebrain ischemia, there were reductions in pyruvate-supported respiration within the first 3 h of recirculation in mitochondria isolated from the dorsolateral striatum (a region in which the majority of neurons are susceptible to ischemia) but not the ischemia-resistant paramedian neocortex. The present study demonstrates that the changes in mitochondrial respiration apparently result from a loss of activity of the pyruvate dehydrogenase complex (PDHC). In mitochondria from the dorsolateral striatum, incubated in the presence of pyruvate and ADP (state 3 conditions) and treated to preserve the phosphorylation state of PDHC, there was no significant change from preischemic activity after 30 min of ischemia or 1 h of recirculation. However, a significant reduction (to 71% of control value) was observed at 3 h of recirculation, and the activity decreased further at 6 and 24 h (to 64 and 43% of control values, respectively). Total PDHC activity in the isolated mitochondria was similarly reduced at 3 h (68% of control values) and 6 h (73% of control values), indicating that the alteration was due to loss or inactivation of the PDHC rather than changes in phosphorylation of the complex. No significant changes were observed in the activity of two other mitochondrial markers, rotenone-sensitive NADH-cytochrome c oxidoreductase and alpha ketoglutarate dehydrogenase. None of the activities of these three enzymes in mitochondria from the paramedian neocortex was significantly affected by ischemia or recirculation. These results (together with previous observations) indicate an early and specific change affecting the PDHC in cells of the dorsolateral striatum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417014 TI - Value of a DNA probe assay (Gen-Probe) compared with that of culture for diagnosis of gonococcal infection. AB - The Gen-Probe PACE 2 system for Neisseria gonorrhoeae (GP), which uses a chemiluminescently labeled DNA probe, was compared with conventional culture as the method of reference. A total of 1,750 specimens were collected from 496 females and 623 males visiting the outpatient clinic of the Sexually Transmitted Diseases Department of the Westeinde Hospital, The Hague, The Netherlands, during the year 1991. The prevalences of gonorrhea culture-positive men and women were 14.9 and 7.7%, respectively. The overall positive rate was 8.7%. Sensitivity, specificity, and positive and negative predictive values of GP were 97.1, 99.1, 90.6, and 99.8%, respectively. A total of 12 of 13 patients with positive GP results and negative cultures may have had a gonococcal infection, a conclusion based on clinical symptoms, positive methylene blue smears, and high relative light unit ratios. The DNA probe test can be useful as a suitable screening and diagnostic test for gonorrheal infection in men and women. An advantage of using this DNA probe technique is that simultaneous testing for Chlamydia trachomatis of the same specimen is possible. We also examined whether (all) rRNA had disappeared after adequate treatment for gonococcal and/or chlamydial infection in 30 patients. None of those positive patients showed a positive result in the DNA probe assay after treatment. PMID- 8417015 TI - Measles virus-specific cellular immunity in patients with vaccine failure. AB - The cytotoxic T-lymphocyte (CTL) response to measles virus (MV) was studied in blood samples from 13 acute- and early convalescent-phase patients with measles infection despite previous vaccination with the live-MV vaccine. MV CTL responses were also measured in six healthy peer controls who had live-MV vaccination during childhood and in five healthy adults who had a remote history of natural measles. All patients recovered from illness without complication. Acute MV infection was diagnosed on the basis of the Centers for Disease Control criteria and by measuring MV-specific immunoglobulin G (IgG) and IgM antibodies. Elevated IgG titers occurred in 80% of the patients at 1 to 2 weeks and in 100% at 4 weeks postinfection. IgM antibodies were detectable in all patient tested and were elevated in 60% of the patients at 1 to 2 weeks postinfection. The MV-specific CTL response was enhanced in 10 of the 13 patients tested, with a mean maximal lysis of 48.5% +/- 13.3%, compared with that of healthy peer controls who had had live-MV vaccinations during childhood (mean lysis, 14.6% +/- 12.9%; n = 6) and healthy adults with a remote history of natural measles (mean, 30.8% +/- 12.2%; n = 5). Three patients had low MV CTL levels at two time points following measles, with a mean lysis of 12% +/- 1.7%. It is concluded that while there is no evidence for a deficiency in the generation of cellular immunity to MV in the majority of patients with MV vaccine failure, a small number of individuals may fail to develop an enhanced T-cell response following infection. PMID- 8417016 TI - Clinical and bacteriological study of nosocomial infections due to Enterobacter aerogenes resistant to imipenem. AB - Enterobacter aerogenes strains resistant to imipenem were isolated in 10 patients, 7 of whom had received imipenem-cilastatin. The strains were differentiated by biotype, antibiotype, and plasmid content. All of the strains overproduced a chromosomal cephalosporinase and lost a major outer membrane protein with a size of about 40 kDa. In 5 of the 10 patients, E. aerogenes strains resistant to extended-spectrum cephalosporin were isolated during the same stay. In three patients, the similarity between the imipenem-susceptible and -resistant strains suggests the occurrence of mutation and reversion in vivo. The combination imipenem-cilastatin has been critically important for use with multiresistant strains of Enterobacter spp., but its use increases the risk of selection of imipenem-resistant strains. PMID- 8417017 TI - Serotyping of European isolates of Chlamydia psittaci from poultry and other birds. AB - A panel of five serovar-specific monoclonal antibodies which distinguish the five known avian serovars of Chlamydia psittaci was used to serotype 45 European avian Chlamydia psittaci isolates. Chlamydial antigen was grown in Buffalo green monkey (BGM) cells or in embryonated chicken eggs and was then inoculated into BGM cells. Serotyping was performed in an indirect immunofluorescence test. The 45 European isolates included 22 isolates from the order Psittaciformes, 9 isolates from the order Columbiformes, 6 isolates from the order Galliformes, 5 isolates from the order Passeriformes, and 3 isolates from the order Anseriformes. All of these were successfully serotyped. No additional serovars were found. One isolate from a duck and two isolates from psittacine birds gave positive immunofluorescences with two monoclonal antibodies considered to be specific for two different serovars. These three isolates were cloned by an agar overlay method. Serotyping of the clones demonstrated that the duck and one psittacine bird each were infected with two different serovars. After cloning, one isolate from a psittacine bird reacted only with serovar A. From these results it was concluded that this serotyping system allows the classification of all isolates tested so far. The results show that similar serovars are prevalent in avian species in Europe and the United States. The results also indicate that birds from a certain order are more susceptible to a distinct serovar. The use of a panel of serovar-specific monoclonal antibodies in the immunofluorescence test provides a reliable method for serotyping avian isolates. Monoclonal antibodies to new avian isolate serovars can easily be added to the panel, which makes the system useful for epidemiological studies. PMID- 8417018 TI - Enzyme-linked immunosorbent assay for detection of human immunoglobulin G to lipopolysaccharide of spotted fever group rickettsiae. AB - An enzyme-linked immunosorbent assay for detecting human immunoglobulin G to spotted fever group rickettsiae was developed and tested. The assay uses proteinase K-resistant material, characteristic of the rickettsial lipopolysaccharides shown to be group specific by immunoblots, as the antigen. The results indicate that the assay provides a sensitive, yet specific, alternative method for diagnosing rickettsial diseases. PMID- 8417019 TI - Evaluation of a rapid enzyme immunoassay for detection of influenza A virus. AB - The Directigen FLU-A enzyme immunoassay for the detection of influenza A virus was compared with direct smear and culture in 211 clinical specimens. The FLU-A enzyme immunoassay proved to be a reliable, rapid screen for influenza A from symptomatic patients and was less dependent on technical expertise for interpretation than were direct smears. PMID- 8417020 TI - Comparison of pooled formalin-preserved fecal specimens with three individual samples for detection of intestinal parasites. AB - Triplicate preserved fecal samples from 265 patients were pooled into single samples, and the recovery rate of intestinal parasites from the pooled samples was compared with that from the individual samples. Parasites were detected in 109 patients when results from the individual samples were used compared with 108 patients when results from the pooled specimens were used. Pooling preserved fecal samples is an efficient and economical procedure for the detection of ova and parasites. PMID- 8417021 TI - Growth and phenotypic characterization of Legionella species on semisolid media made with washed agar. AB - Legionella pneumophila (and 28 Legionella species) grew efficiently on charcoal free, buffered yeast extract medium made with washed agar and without apparent loss of infectivity for U937 cells. Because charcoal-free, buffered yeast extract is transparent, it is a suitable base for indicator media and pigment detection. In standard media, charcoal apparently prevents agar contaminants from inhibiting Legionella growth. PMID- 8417022 TI - Detection of Francisella tularensis in blood by polymerase chain reaction. AB - We developed a polymerase chain reaction-based assay for Francisella tularensis which we evaluated by using spiked blood samples and experimentally infected mice. The assay detected both type A and type B F. tularensis at levels equivalent to one CFU/microliter of spiked blood. Results from polymerase chain reaction-based assay of limiting dilutions of blood from mice infected with the live vaccine strain agreed closely with results from blood culture. PMID- 8417023 TI - Sucrose-positive Edwardsiella tarda mimicking a biogroup 1 strain isolated from a patient with cholelithiasis. AB - An unusual strain of Edwardsiella tarda mimicking a biogroup 1 isolate was recovered in a mixed infection from a woman suffering from cholelithiasis. Rare biochemical characteristics (e.g., H2S negativity) originally detected were related to an unusual biochemical property in this species, sucrose fermentation; other points of interest regarding this strain included the site of isolation (bile) and the failure of this isolate to produce many of the in vitro virulence markers associated with E. tarda. PMID- 8417024 TI - Use of electrophoretic polymorphisms of esterases for differentiation of Clostridium argentinense strains. AB - Esterase electrophoresis was used to study 10 strains of Clostridium argentinense, including 7 toxigenic and 3 nontoxigenic strains. On the basis of the electrophoretic mobilities and hydrolytic specificities toward five synthetic substrates, different esterase profiles could be defined for almost all strains, revealing the heterogeneity of bacterial clones. Therefore, electrophoretic polymorphism of esterases can be used for differentiation of C. argentinense in population genetic or epidemiological studies. PMID- 8417025 TI - Human intestinal spirochetes are distinct from Serpulina hyodysenteriae. AB - Twenty-nine intestinal spirochetes isolated from Australian aboriginal children and six strains from Italian adults (HRM1, -2, -4, -5, -7, and -14) were genetically examined at 15 enzyme loci by using multilocus enzyme electrophoresis. Results were compared with those previously obtained for 188 porcine intestinal spirochetes. DNA from human strain HRM7 and porcine strain Serpulina hyodysenteriae P18A were also radioactively labeled and hybridized with DNA from 12 other human and porcine intestinal spirochetes. Both the multilocus enzyme electrophoresis and hybridization techniques demonstrated that the human spirochetes were not S. hyodysenteriae. They belonged to another distinct genetic group of spirochetes that included P43/6/78, the bacterium recovered from the first recorded case of porcine intestinal spirochetosis. Bacteria in this distinct group also differed from Serpulina spp. in possessing only four, five, or occasionally six axial filaments, being slightly thinner, and having more pointed ends. These findings add further weight to the possibility that human intestinal spirochetes may act as enteric pathogens. PMID- 8417026 TI - Growth of Helicobacter pylori in media containing cyclodextrins. AB - We show that solid and liquid media, supplemented only with cyclodextrins and free of blood and its derivatives, support the growth of Helicobacter pylori. These media can be used for primary isolation of the bacteria from biopsy samples, routine laboratory growth, and large-scale industrial fermentation. PMID- 8417027 TI - Rapid immunodot technique for identifying Bordetella pertussis. AB - We developed and evaluated a rapid test with monoclonal antibodies to identify cultures of Bordetella pertussis. Samples of 5 microliters of cells suspended in formalin-saline were dried onto a nitrocellulose disk. The disk was placed in a filtration device, and 5-microliters volumes of murine monoclonal antibody directed against B. pertussis lipooligosaccharide and peroxidase conjugate were added consecutively, with washing after each addition. The disk was removed and immersed in peroxidase substrate solution. All of 66 B. pertussis isolates confirmed by direct fluorescent-antibody assay were correctly identified by using four different monoclonal antibodies. One of the monoclonal antibodies did not react with over 20 bacterial species tested, including other Bordetella, Acinetobacter, Haemophilus, Moraxella, Mycobacterium, Neisseria, and Staphylococcus spp. This technique detected > or = 2 micrograms of lipooligosaccharide per ml or > or = 5 x 10(8) B. pertussis cells per ml. This rapid procedure used small amounts of reagents, needed less equipment, and was less subjective and more specific than the direct fluorescent-antibody assay. PMID- 8417028 TI - Helicobacter pylori serology. PMID- 8417029 TI - Biochemical properties and fatty acid composition of Mycobacterium haemophilum: study of 16 isolates from Australian patients. AB - The biochemical properties and fatty acid compositions of 16 strains of Mycobacterium haemophilum from Australian patients were studied. The strains proved to be indistinguishable from each other but could readily be differentiated from other slowly growing mycobacteria with similar cultural features. Mycolic acid analyses revealed the presence of alpha-, methoxy-, and ketomycolates. The fatty acid composition supports the validity of the fact that M. haemophilum is a distinct species. The fatty acid composition was consistent among the 16 strains, but it was unusual in that there was some resemblance to the fatty acid composition of M. leprae. The wide range of pHs (5.4 to 7.4) that supported growth of M. haemophilum on artificial medium is in keeping with suggestions that M. haemophilum exists in an environmental habitat. PMID- 8417030 TI - Genetic similarity of Candida albicans strains from vaginitis patients and their partners. AB - The moderately repetitive sequence Ca3 was used to fingerprint strains of Candida albicans isolated from vulvovaginal infections of 10 women and strains isolated from their male partners. The Dendron software package was then used to compare the DNA fingerprints of these strains with those of vaginal commensals from women from the same geographical locale, vaginal commensals from women from a different geographical locale, and commensals from male partners of asymptomatic women from the same geographical locale. The results demonstrate that, in the majority of cases (8 of 10), strains from symptomatic patients and their partners are either identical or more similar to each other than to other strains, infecting strains do not represent a group genetically distinguishable from vaginal commensal isolates from women from the same geographical locale, and both infecting strains and commensals from individuals in the test locale can be distinguished from commensals obtained in another geographical locale. The results also suggest that women with vaginal infections are responsible for strain replacement in their male partners. PMID- 8417031 TI - Do temperature-sensitive auxotrophs of Escherichia coli have special virulence? AB - To determine whether temperature (42 degrees C)-sensitive auxotrophs of Escherichia coli have special virulence properties (W. D. Welch, D. Kitts, H. S. Moyed, and L. D. Thrupp, J. Clin. Microbiol. 13:606-608, 1981), we examined 301 strains isolated from patients with bacteremia or acute cystitis and from the stools of healthy subjects. Of these strains, 49.5% grew at 42 degrees C without supplements, 39.2% required a nutritional supplement, and 11.3% failed to grow even with selected nutrients. Nicotinamide restored growth for 35.2% of strains at either 37 or 42 degrees C. Some of strains required methionine, glutamic, aspartic, and amino acid mixtures or NaCl for growth at 42 degrees C. Temperature sensitive strains were significantly more abundant in isolates from blood and urine than in stool, but temperature-sensitive auxotrophs were isolated at about the same frequency from each site. There were no discernible clonal patterns, by serotype, among of the nicotinamide-requiring temperature-sensitive auxotrophs. Resistance to trimethoprim-sulfamethoxazole was associated with ability to grow at 42 degrees C. This was not observed with any other antimicrobial drug. Temperature-sensitive strains are a heterogenous group. The relationship of temperature-sensitive auxotrophy to virulence is uncertain. PMID- 8417032 TI - Comparison of nasopharyngeal aspirates with swabs for culture of Bordetella pertussis. AB - Nasopharyngeal samples were collected from 117 children by aspiration from one nostril and by swab from the other one and cultured for Bordetella pertussis. Among 33 culture-positive specimens, there were 30 positive aspirates and 26 positive swab specimens. Aspirates are easily divided and saved for investigations with other assays which may further improve diagnostic sensitivity. As the aspiration technique was also preferred by nurses and parents, it was recommended and chosen for a planned pertussis vaccine efficacy trial to take place in Sweden from 1992 to 1995. PMID- 8417033 TI - Development and use of species-specific oligonucleotide probes for differentiation of Streptococcus uberis and Streptococcus parauberis. AB - Oligonucleotide probes specific for 16S rRNA and capable of differentiating Streptococcus uberis and S. parauberis from each other and other esculin hydrolyzing streptococci were developed. Use of a mini-RNA extraction technique for gram-positive cocci associated with bovine mastitis has allowed the probes to be used for identification of esculin-hydrolyzing streptococci from two dairy herds at the Institute for Animal Health, Compton, United Kingdom. One hundred seventy-nine of 206 isolates were identified as S. uberis, 3 were identified as S. parauberis, and 24 were not identified. Isolates not identified by the probes were tested biochemically and found to be mainly Enterococcus faecium, E. faecalis, or S. bovis. PMID- 8417034 TI - Detection of Mycobacterium tuberculosis in clinical samples by two-step polymerase chain reaction and nonisotopic hybridization methods. AB - Detection of Mycobacterium tuberculosis in clinical specimens by the polymerase chain reaction (PCR) was compared with detection by culture. A 317-bp segment within the M. tuberculosis-specific insertion sequence IS6110 was amplified. The detection limit of the PCR assay for cultured mycobacteria was 50 cells per reaction by ethidium bromide-stained agarose gel electrophoresis and 5 cells per reaction by hybridization with an oligonucleotide probe conjugated with either digoxigenin or alkaline phosphatase (AP). This sensitivity was reduced fivefold in sputum specimens seeded with M. tuberculosis. Seventy-six clinical specimens were amplified and examined by the three detection methods. Both the digoxigenin and AP procedures were found to be more sensitive than agarose gel electrophoresis, but they were occasionally associated with a high background. An additional 308 specimens were examined only by agarose gel electrophoresis and the AP procedure. Of 71 specimens found to contain M. tuberculosis, amplified products were detected from 56 (79%) samples by agarose gel electrophoresis and/or the AP procedure. Of the additional 313 specimens that were culture negative for M. tuberculosis, 19 (6%) had amplified products detectable by agarose gel electrophoresis and/or the AP procedure. Compared with culture, PCR showed sensitivities and specificities of 55 and 98%, respectively, for agarose gel electrophoresis and 74 and 95%, respectively, for the AP procedure. Despite this low sensitivity, a rapid positive PCR result was accurate and clinically useful. PMID- 8417035 TI - Prevalence of dental caries in 4- to 5-year-old children partly explained by presence of salivary mutans streptococci. AB - The correlation between dental caries and the number of oral mutans group streptococci (ms) present has been shown to be weak. The aim of this investigation was to study associations between caries experience (decayed, missing, and filled surfaces [dmfs]) and the number of ms in stimulated saliva, with emphasis on the level of disease and the confounding effect of regular intake of sweets, the presence of salivary lactobacilli, and oral hygiene. In some 2,700 4- to 5-year-old South African children of different ethnic origins, caries was diagnosed on the basis of World Health Organization criteria and saliva samples were analyzed for ms after cultivation on mitis salivarius bacitracin agar and for lactobacilli by using the Dentocult kit. Oral hygiene was scored on the basis of the Greene and Vermillion simplified debris index, while data on intake of sweets were derived from extensive interviews. Pearson's coefficient of correlation was computed, and multiple regression analysis was performed to correct for confounding factors. The distribution of the children in the eight caries classes was strongly associated with the ms class (P < 0.001), with those in the lower ms classes generally having low dmfs scores and those in the higher ms classes having dmfs scores distributed over the whole range. The r value for the two variables was 0.25 for the total material; this was reduced to 0.18 by correction for confounding factors. The corresponding values for children with caries were 0.21 and 0.17, for those in the 1 to 6 dmfs interval they were 0.07 and 0.03, and for those in the 7 to 81 dmfs interval they were 0.16 and 0.14. The data imply that the explanatory values for ms, those for the lower caries interval not counted, ranged from 6 to 2%. The unexpected results for children with caries might be due to their distribution pattern. It is concluded that there is a need for reevaluation of ms as a risk factor in dental caries. PMID- 8417036 TI - Comparison of two antigen assays for rapid intrapartum detection of vaginal group B streptococcal colonization. AB - As part of a clinical investigation evaluating the efficacy of intrapartum antigen detection for screening for heavy vaginal colonization with group B streptococci (GBS), we compared the performance of modified Bactigen and Directigen GBS latex particle agglutination (LPA) kits. Paired vaginal swabs obtained from women in labor were rapidly transported to the laboratory and used for culturing (both swabs) and LPA testing (one swab by each method). GBS growth was estimated semiquantitatively and further designated as light or heavy growth. Performance specifications for each method were determined by comparing LPA and culture results from the same swab. A total of 4,251 paired swabs were evaluated during the study period. The performance specifications for detecting GBS growth of any degree for Bactigen and Directigen, respectively, were as follows: sensitivity, 20 and 24%; specificity, 99 and 99%. The performance specifications for heavy colonization for Bactigen and Directigen, respectively, were as follows: sensitivity, 57 and 62%; specificity, 99 and 99%. Neither LPA kit was a sensitive indicator of vaginal colonization with GBS or neonatal infection. PMID- 8417037 TI - Serodiagnosis of toxoplasmosis by using a recombinant form of the 54-kilodalton rhoptry antigen expressed in Escherichia coli. AB - A 330-residue carboxy-terminal antigenic fragment of the Toxoplasma gondii 54-kDa rhoptry protein (ROP2) was expressed in Escherichia coli as a fusion polypeptide containing a 48-amino-acid sequence derived from phage lambda protein Cro and E. coli protein LacI followed by six consecutive histidyl residues. Metal chelate affinity chromatography provided an easy way to isolate the recombinant product in a highly purified form (> 95%). When this material was used to develop an immunoglobulin G (IgG) enzyme-linked immunosorbent assay for diagnosis of toxoplasmosis, the test reached a sensitivity of 89%. The sensitivity of the assay was similar whether the sera contained T. gondii-specific IgM or were devoid of such IgM. It was also found that ROP2 is a conserved antigen since antibodies against the recombinant antigen could be detected in mice experimentally infected with 11 independent T. gondii isolates originating from infected human tissues tested. Thus, the 54-kDa rhoptry antigen could advantageously complement other previously described T. gondii antigens for the development of more-sensitive and more-informative recombinant antigen-based tests for toxoplasmosis diagnosis. PMID- 8417038 TI - Evaluation of an enzyme immunoassay for detection of cryptococcal capsular polysaccharide antigen in serum and cerebrospinal fluid. AB - The Premier enzyme immunoassay (Meridian Diagnostics, Inc., Cincinnati, Ohio) was compared with a latex agglutination assay (CALAS; Meridian) for the ability to detect cryptococcal capsular polysaccharide antigen (CrAg) in serum and cerebrospinal fluid (CSF). A total of 594 specimens (471 serum samples and 123 CSF samples) obtained from 430 patients, most of whom were at risk for or had AIDS, were tested in parallel by both systems. Both tests were independently evaluated for their ability to (i) detect CrAg when used as a screening test and (ii) quantitate the CrAg present when used as a titration assay. Chart review to assess clinical outcome after the time of specimen collection was conducted for all patients. When both assays were used as screening assays, 103 serum samples and 18 CSF samples were positive and 356 serum samples and 104 CSF specimens were negative by both assays (97.8% concordance). Thirteen specimens (12 serum samples, 1 CSF sample) gave discrepant screening results. When the tests were used as semiquantitative assays for titer determinations, the CrAg titers determined by the enzyme immunoassay were generally higher than those obtained with the latex agglutination assay. In summary, results obtained with the enzyme immunoassay correlated well with those obtained with the latex agglutination test for screening for the presence of CrAg and for determining the titer of CrAg in serum or CSF. PMID- 8417039 TI - A controlled evaluation of computer assisted training simulations in geriatric dentistry. AB - The increasing number of geriatric dental patients and the development of treatment plans that require consideration of complex psychosocial, socioeconomic, and medical/medication factors necessitates a change in the traditional teaching of treatment planning. A computer-assisted instructional program was developed to simulate the dentist-geriatric patient interaction and to train students in clinical decision making for the geriatric patient. This study compared the effects of this program with a more traditional approach based on readings from the geriatric dentistry literature. Twenty third-year dental students were matched on grade point average and randomly assigned to the computer or literature-based groups. They were pretested using a clinical analogue of a geriatric patient and then instructed to use either the computer or literature-based educational units. The students were then post-tested on a second clinical analogue. Students performed similarly at pretest. At post-test, both the computer and literature-based subjects were found to have acquired significantly greater evaluation points, to make fewer errors, and to design more involved treatment plans than at pretest. Comparison of computer and literature based subjects' performance revealed no significant main or interaction effects regarding type of educational unit used. Consistent nonsignificant trends were noted with the computer-based subjects out-performing literature-based subjects on each outcome variable. In addition, the computer-based subjects had more positive feelings about the educational unit than the literature-based subjects at a statistically significant level. These findings suggest that the computer program is an effective alternative method for developing clinical decision skills in students treating geriatric patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417040 TI - Responsibility for teaching pain control in U.S. dental schools. PMID- 8417041 TI - CPR requirements for dental schools and state licensure. PMID- 8417042 TI - Human cadaver material in preclinical oral surgery. PMID- 8417043 TI - Curriculum guidelines for removable prosthodontics. American Association of Dental Schools Section on Removable Prosthodontics. PMID- 8417044 TI - Curriculum guidelines for fixed prosthodontics. American Association of Dental Schools Section on Fixed Prosthodontics. PMID- 8417045 TI - The AADS thrust to an international flavor. PMID- 8417046 TI - Curriculum guidelines in implant dentistry for general practice residency and advanced education in general dentistry programs. American Association of Dental Schools. PMID- 8417047 TI - ADEE--from travelers' club to pressure group. PMID- 8417048 TI - Percutaneous balloon pericardiotomy for the treatment of cardiac tamponade and large pericardial effusions: description of technique and report of the first 50 cases. AB - OBJECTIVES: This study describes the technique, clinical characteristics and results of the first 50 patients undergoing percutaneous balloon pericardiotomy as part of a multicenter registry. BACKGROUND: Percutaneous balloon pericardiotomy involves the use of a percutaneous balloon dilating catheter to create a nonsurgical pericardial window. METHODS: Patients eligible for percutaneous balloon pericardiotomy had either cardiac tamponade (n = 36) or a moderate to large pericardial effusion (n = 14). In addition to clinical follow up, serial echocardiograms and chest X-ray films were obtained. RESULTS: The procedure was considered successful in 46 patients after a mean follow-up period of 3.6 +/- 3.3 months. Two patients required an early operation, one for bleeding from a pericardial vessel and one for persistent pericardial catheter drainage. Two patients required a late operation for recurrent tamponade. Minor complications of the procedure included fever in 6 of the first 37 patients (studied before the prophylactic use of antibiotic agents), thoracentesis or chest tube placement in 8 and a small spontaneously resolving pneumothorax in 2. Despite the short-term success of this procedure, the long-term prognosis of the 44 patients with malignant pericardial disease remained poor (mean survival time 3.3 +/- 3.1 months). CONCLUSIONS: Percutaneous balloon pericardiotomy is successful in helping to manage large pericardial effusions, particularly in patients with a malignant condition. It may become the preferred treatment to avoid a more invasive procedure for patients with pericardial effusion and a limited life expectancy. PMID- 8417049 TI - A prospective randomized comparison of direct current and radiofrequency ablation of the atrioventricular junction. AB - OBJECTIVES: The purpose of this study was to compare direct current and radiofrequency ablation of the atrioventricular (AV) junction in a prospective randomized fashion. BACKGROUND: Catheter ablation of the AV junction can be performed using either direct current shocks or radiofrequency energy. To date, these two techniques have never been compared prospectively or in a randomized study. METHODS: Forty patients with drug-refractory uncontrolled atrial fibrillation-flutter (38 patients) or inappropriate sinus tachycardia (2 patients) were randomly assigned to undergo direct current ablation (20 patients) using up to four shocks of 200 to 300 J or radiofrequency ablation (20 patients) using up to 15 applications of 16 to 25 W for 30 s. If complete AV block was not successfully induced, the ablation procedure was repeated using the alternate type of energy. A rate-responsive ventricular pacemaker was implanted in each patient. The intrinsic escape rhythm was evaluated 15 min, 2 days and 3, 6 and 12 months after ablation. RESULTS: Persistent complete AV block was successfully induced during the first ablation session in 13 (65%) of 20 patients randomly assigned to undergo direct current ablation, compared with 19 (95%) of 20 patients randomly assigned to undergo radiofrequency ablation (p < 0.05). Each patient whose first ablation attempt failed had a successful outcome with the alternate type of energy. The overall efficacy of radiofrequency ablation (26 [96%] of 27 patients) was significantly greater than that of direct current ablation (14 [67%] of 21 patients, p < 0.01). The duration of the direct current and radiofrequency ablation sessions did not differ significantly. The mean peak plasma creatine kinase MB fraction concentration was significantly higher after direct current ablation (58 +/- 29 IU/liter) than after radiofrequency ablation (2 +/- 2 IU/liter) (p < 0.001). An escape rhythm was present 15 min after ablation in an equal proportion of patients undergoing direct current and radiofrequency ablation (78% and 85%, respectively, p = 0.6). An escape rhythm was present in all patients 3, 6 and 12 months after ablation. The mean escape rhythm cycle length 15 min after direct current ablation (2,074 +/- 677 ms) was significantly longer than that 15 min after radiofrequency ablation (1,460 +/- 294 ms) (p < 0.05); however, the mean escape rhythm cycle lengths did not differ significantly at 2 days or 3, 6 or 12 months after ablation. Immediate arrhythmic complications did not occur after either procedure. One patient died suddenly 6.5 months after direct current ablation. CONCLUSIONS: Radiofrequency ablation of the AV junction is more efficacious and safer than direct current ablation and should be the preferred method for inducing complete AV block in patients who are appropriate candidates for ablation of AV conduction. PMID- 8417050 TI - Syncope in advanced heart failure: high risk of sudden death regardless of origin of syncope. AB - OBJECTIVES: The purpose of this study was to assess the importance of syncope as a warning sign for sudden death in advanced heart failure and to determine the relative importance of cardiac syncope and syncope from other causes. BACKGROUND: Despite remarkable advances in the pharmacologic approach to advanced heart failure, 20% to 40% of patients with advanced heart failure will die each year. In such patients, the relation between sudden death and the etiology of syncope has not been evaluated. METHODS: The relation of syncope to sudden death was evaluated in 491 consecutive patients with advanced heart failure (New York Heart Association functional class III or IV), no history of cardiac arrest and a mean left ventricular ejection fraction of 0.20 +/- 0.07. Patients were evaluated for the presence and origin of syncope. The severity of heart failure was assessed from serum sodium levels, ejection fraction, functional class and echocardiographic and hemodynamic variables. RESULTS: Sixty patients (12%) had a history of syncope; the condition had a cardiac origin in 29 (48%) and was due to other causes in 31 (52%). The origin of heart failure was coronary artery disease in 234 patients (48%) and dilated cardiomyopathy in 253 (51%) and its severity was similar in patients with and without syncope. During a mean follow-up interval of 365 +/- 419 days, 69 patients (14%) died suddenly and 66 patients (13%) died of progressive heart failure. The actuarial incidence of sudden death by 1 year was significantly greater in patients with (45%) than in those without (12%, p < 0.00001) syncope. In the Cox proportional hazards model, syncope predicted sudden death independent of atrial fibrillation, serum sodium, cardiac index, angiotensin-converting enzyme inhibition and patient age. The actuarial risk of sudden death by 1 year was similarly high in patients with either cardiac syncope or syncope from other causes (49% vs. 39%, p = NS). CONCLUSIONS: Patients with advanced heart failure are at especially high risk for sudden death regardless of the etiology of syncope. PMID- 8417051 TI - Nonatherosclerotic narrowing of the atrioventricular node artery and sudden death. AB - OBJECTIVES: This study was undertaken to determine whether thickening of the atrioventricular (AV) node artery is a cause of sudden cardiac death. BACKGROUND: Thickening of the AV node artery has been implicated as a cause of sudden death primarily on the basis of case reports. Few pathologic studies have compared subjects who died of sudden cardiac death with normal control subjects who died traumatically. METHODS: The AV node artery in 27 patients with unexplained sudden cardiac death (mean age 24.8 +/- 7.4 years) was compared with that in 17 control subjects who died traumatically (mean age 25.6 +/- 7.0 years). No anatomic cause of death was found at autopsy in the subjects with sudden death, all of whom died of presumed cardiac arrhythmias. The conduction system of all hearts was studied by semiserial sections and Movat pentachrome stains. At the point of greatest narrowing of the AV node artery, the outer circumference and lumen outline were traced by computerized morphometry, the ratio of outer vessel area to lumen area was calculated and the histopathologic changes were noted. RESULTS: The rank-sum of ratios was significantly greater in the sudden death group than in the control group (p = 0.031, Wilcoxon rank-sum/Mann-Whitney statistic). A dysplastic AV node artery with significant acid mucopolysaccharide deposition was seen almost exclusively in the sudden death group (12 of 27 vs. 1 of 17, p = 0.006). In 10 subjects with sudden death a dysplastic AV node artery was narrowed > 2 SD over the control value; half of this subgroup died during exercise and one third had a family history of sudden unexplained cardiac death. CONCLUSIONS: Dysplasia of the AV node artery may contribute to death in a substantial portion of patients with unexplained sudden death, and such death is often associated with exercise and a family history of unexplained sudden death. PMID- 8417052 TI - Postoperative pulmonary flow dynamics after Fontan surgery: assessment with nuclear magnetic resonance velocity mapping. AB - OBJECTIVES: This study was performed to assess the value of nuclear magnetic resonance (NMR) velocity mapping for the measurement of pulmonary blood flow after Fontan surgery. BACKGROUND: Echocardiographic studies of pulmonary flow after Fontan surgery are not always satisfactory. The newly developed technique of NMR velocity mapping may contribute to the elucidation of the Fontan circulation. METHODS: At frequent intervals during the cardiac cycle, forward and backward flow volumes in the pulmonary arteries of nine volunteers were measured, summed and compared with right ventricular stroke volume to validate the velocity mapping technique. In 14 patients after Fontan surgery, assessment of pulmonary flow volumes enabled the evaluation of atriopulmonary and atrioventricular (AV) Fontan connections. The findings were correlated with precordial echocardiography. RESULTS: Validation of the NMR technique, obtained from volunteer experiments, showed a high correlation (r = 0.97) between right ventricular stroke volume and volumetric pulmonary stroke flow. In all patients with an atriopulmonary Fontan connection (n = 8), forward flow in the pulmonary artery was biphasic, similar to normal venous flow. Monophasic systolic pulmonary flow curves indicating right ventricle-dependent pulmonary blood flow were found in three of six patients with an AV Fontan connection. In the remaining three patients, the pulmonary flow pattern did not reflect right ventricular contraction. Measurement of flow velocity alone may give a false impression of forward flow and thus of right ventricular contribution. Pulmonary regurgitation was demonstrated in six of eight patients with an atriopulmonary connection. CONCLUSIONS: Nuclear magnetic resonance velocity mapping provides accurate and valuable information on pulmonary flow volume and velocity after Fontan surgery. The success of AV Fontan surgery can be deduced from the presence of a monophasic systolic pulmonary flow pattern as demonstrated by NMR velocity mapping. With NMR flow volume analysis, substantial pulmonary regurgitation occurring after atriopulmonary Fontan surgery can be measured. PMID- 8417053 TI - Five-year follow-up after balloon pulmonary valvuloplasty. AB - OBJECTIVES: The aim of this study was to assess results 5 years after balloon pulmonary valvuloplasty. BACKGROUND: Since the technique of balloon pulmonary valvuloplasty was first reported in 1982, it has become the treatment of choice for pulmonary valve stenosis. In contrast to surgical valvotomy, the long-term outcome after balloon pulmonary valvuloplasty is unknown. METHODS: We reviewed the findings in 34 patients 5.2 +/- 0.8 (mean +/- SD) years after balloon pulmonary valvuloplasty: 27 with isolated pulmonary stenosis, 5 with Noonan syndrome and 2 with previous surgical valvotomy. In eight patients (three with Noonan syndrome), a second balloon valvuloplasty was the index procedure for analysis. RESULTS: The transpulmonary gradient (mm Hg) was 74 +/- 34 before balloon pulmonary valvuloplasty, 36 +/- 26 immediately after, 22 +/- 9 at cardiac catheterization in 29 patients 6 +/- 0.6 months later and 19 +/- 10 by Doppler study at 5 years. At 5 years 26 patients (group A) had a residual gradient of < or = 20 mm Hg; the remaining 8 (group B) had a gradient of > 20 mm Hg. Four group B patients had Noonan syndrome (p = 0.01). Balloon/pulmonary valve diameter ratio was larger for group A patients than for group B patients with isolated pulmonary stenosis (1.20 +/- 0.10 vs. 1.00 +/- 0.07, p = 0.005); larger balloons were used in group B patients with Noonan syndrome (1.30 +/- 0.10). Group A patients were more likely than group B patients to have significant pulmonary incompetence (6 of 24 vs. 0 of 8) and had a greater right ventricle/left ventricle long-axis diastolic dimension ratio (0.47 +/- 0.10 vs. 0.35 +/- 0.04, p = 0.05). In the subgroup of five patients with Noonan syndrome and two with prior surgical valvotomy, the transpulmonary gradient was reduced from 74 +/- 24 mm Hg before balloon valvuloplasty to 23 +/- 12 mm Hg at 5 years. In addition, two patients with isolated pulmonary valve stenosis had pulmonary valve dysplasia by angiographic criteria: transpulmonary gradients of 85 and 56 mm Hg were reduced to 20 and 11 mm Hg, respectively, at 5 years. CONCLUSIONS: Relief of obstruction persists at 5 years especially if oversized balloons are used. Acceptable results can be obtained in patients with a dysplastic valve. More complete relief of right ventricular outflow gradient is associated with increased right ventricular dimension, probably because more pulmonary incompetence is induced. This is well tolerated at 5 years but may be important in the longer term. PMID- 8417054 TI - Solitary coronary ostium in the aorta in the absence of other major congenital cardiovascular anomalies. AB - OBJECTIVES: This study examines the distribution patterns and the clinical significance of the "solitary coronary ostium" in the aorta in the absence of other major congenital cardiovascular anomalies. BACKGROUND: Ogden in 1970 classified "single coronary artery" into 14 basic distribution patterns. Since then, other patterns of distribution of single coronary artery have been recognized. Distinction has also been made between the cases with and without other major congenital cardiovascular anomalies or coronary artery atresia and those without these additional abnormalities. Single coronary artery has been generally considered to be a benign clinical entity. METHODS: This study describes 10 cases of single coronary artery at necropsy and reviews 87 previously reported cases, 35 diagnosed at necropsy and 52 by coronary angiography. RESULTS: We classified single coronary artery into 20 categories on the basis of the location of the solitary coronary ostium, the presence or absence of an aberrant-coursing coronary artery and the course taken by the aberrant-coursing coronary artery. When atherosclerotic coronary artery disease was absent, 15% (8 of 53) of the patients reviewed with single coronary artery had myocardial ischemia as a direct consequence of the coronary anomaly. CONCLUSIONS: The anatomic classification presented is useful from both clinical and surgical viewpoints. This comprehensive classification of this rare anomaly facilitates description of the various distribution patterns of single coronary artery and their clinical significance. PMID- 8417055 TI - Transesophageal echocardiographically detected atherosclerotic aortic plaque is a marker for coronary artery disease. AB - OBJECTIVES: The aim of this study was to test the hypothesis that atherosclerotic plaque in the thoracic aorta detected by transesophageal echocardiography is a marker for coronary artery disease. BACKGROUND: Previous pathologic and roentgenographic studies have suggested a relation between aortic plaque and coronary artery disease but have lacked clinical utility. METHODS: We performed transesophageal echocardiography on 61 patients (30 women and 31 men aged 22 to 83 years [mean 60 +/- 14]) who had previously undergone cardiac catheterization with coronary angiography. The clinical indications for angiography were angina (n = 26), valvular heart disease (n = 17), positive noninvasive evaluation for ischemia without angina (n = 6), postmyocardial infarction (n = 5), familial hypercholesterolemia (n = 4), coronary cameral fistula (n = 1), atrial myxoma (n = 1) and suspected aortic dissection (n = 1). All patients underwent transesophageal echocardiography with imaging of the thoracic aorta. The criteria used to diagnose atherosclerotic plaque on transesophageal echocardiography were the presence of linear or focal increased echo-density with lumen irregularity and thickening or calcification of the aortic intima. RESULTS: In 41 of the 61 patients, obstructive coronary artery disease was detected by angiography in at least one vessel (> 50% left main coronary artery stenosis or > 70% stenosis in the left anterior descending, right coronary or left circumflex artery distribution). In 37 of the 41, atherosclerotic plaque was detected in the thoracic aorta by transesophageal echocardiography. Twenty of the 61 patients had normal coronary angiographic findings or nonobstructive lumen irregularities. In 2 of these 20 patients, plaque was detected in the thoracic aorta on transesophageal echocardiography. The presence of aortic plaque on transesophageal study had a sensitivity of 90% and a specificity of 90% for angiographically proved obstructive coronary artery disease. The positive predictive value of aortic plaque for obstructive coronary artery disease was 95% and the negative predictive value was 82%. CONCLUSIONS: The detection of atherosclerotic plaque in the thoracic aorta by transesophageal echocardiography appears to be a marker for the identification of obstructive coronary artery disease and deserves further investigation. PMID- 8417056 TI - Generalized model of restenosis after conventional balloon angioplasty, stenting and directional atherectomy. AB - OBJECTIVES: This study was designed to extend the results of a quantitative model originally developed for restenosis after stenting or atherectomy to include restenosis after conventional balloon angioplasty. BACKGROUND: We have previously described a continuous regression model that explains late (6-month) lumen narrowing as the difference between the immediate gain and the subsequent normally distributed late loss in lumen diameter after Palmaz-Schatz stenting or directional atherectomy. METHODS: Lumen diameter was measured immediately before and after coronary intervention on 524 consecutive lesions including those treated by Palmaz-Schatz stenting (102), directional atherectomy (134) and conventional balloon angioplasty (288). Of these lesions, 475 (91%) underwent follow-up angiography 3 to 6 months after treatment. The immediate increase in lumen diameter produced by the intervention (immediate gain) and the subsequent reduction in lumen diameter between the time of intervention to follow-up angiography (late loss) were examined. Association between demographic or angiographic variables and continuous measures of restenosis (late lumen diameter or late percent stenosis) was tested with linear regression techniques; a traditional binary measure of restenosis (late diameter stenosis > or = 50%) was evaluated with logistic regression analysis. RESULTS: Regression models relating late lumen diameter to the immediate lumen result were successfully fitted to all segments studied. According to these models, three indexes of restenosis (late lumen diameter, late percent stenosis and binary restenosis) were found to depend solely on the immediate lumen diameter after the procedure and the immediate residual percent stenosis, but not on the specific intervention used. Moreover, the late loss in lumen diameter was found to vary directly with the immediate gain provided by an intervention, and the "loss index" (a measure that corrects for differences in immediate gain) was uniform among all three interventions. CONCLUSIONS: The quantitative model originally developed for restenosis after stenting or atherectomy may thus be generalized to include conventional balloon angioplasty. It shows that the apparent differences in restenosis among the three interventions studied are due solely to differences in the immediate result provided and not to differences in the behavior of subsequent late loss. Moreover, although the late loss in lumen diameter was found to correlate with differences in the immediate gain provided by an intervention, the "loss index" (a measure that corrects for differences in acute gain) was uniform across all three interventions. It is thus the immediate result (and not the procedure used to obtain that result) that determines late outcome after coronary intervention. PMID- 8417057 TI - Survival 15 to 20 years after coronary bypass surgery for angina. AB - OBJECTIVES: The aim of this study was to determine the 15- to 20-year outcome of coronary bypass surgery in patients with angina. BACKGROUND: Coronary bypass surgery has been performed for > 20 years; we need to know the expected outcome of a very long-term follow-up. METHODS: Using actuarial techniques, we determined the outcome of coronary bypass surgery performed for chronic stable and unstable angina in 7,529 patients from 1969 to 1988. RESULTS: The 5-, 10-, 15- and 20-year survival rates (mean +/- SE) were 88 +/- 1, 73 +/- 1, 53 +/- 1 and 38 +/- 3%, respectively, for the whole group. Compared with patients operated on in 1974 to 1988 (n = 7,026), patients operated on in 1969 to 1973 (n = 503) were younger and had less coronary artery disease but had a higher operative mortality rate and a shorter long-term survival time; 15- and 20-year survival of the 1969 to 1973 cohort was 47 +/- 2% and 33 +/- 3%, respectively. The 1974 to 1988 cohort of patients had a 2.1% operative mortality rate and a 10- and 15-year survival probability of 74 +/- 1% and 55 +/- 2%, respectively. For 2,128 patients with "normal" left ventricular function, the 10- and 15-year survival probability was 82 +/- 1% and 64 +/- 3%, respectively, and for 2,413 patients with "abnormal" left ventricular function, it was 66 +/- 1% and 47 +/- 3%, respectively (p < 0.0001); for men it was 74 +/- 1% and 56 +/- 2%, respectively, and for women, 70 +/- 2% and 52 +/- 5%, respectively, p < 0.05. The actuarial percentages of reoperation and myocardial infarction at 15 years were 33 +/- 2% and 26 +/- 2%, respectively; these values did not differ significantly between men and women. There was a significant (p < 0.001) difference between men and women in angina status; 81% of the men versus 74% of the women had no angina or mild angina at the most recent follow-up study. CONCLUSIONS: Coronary bypass surgery is an effective form of therapy for angina (for 15 to 20 years) in both men and women. PMID- 8417058 TI - Transformation of coronary artery aneurysm to obstructive lesion and the role of collateral vessels in myocardial perfusion in patients with Kawasaki disease. AB - OBJECTIVES: The aim of this study was to examine the transformation of coronary artery aneurysms to obstructive lesions and to assess the role of collateral vessels in patients with Kawasaki disease. BACKGROUND: Coronary artery aneurysms, especially giant aneurysms, are known to become obstructive lesions in patients with Kawasaki disease. However, the process of transformation is not yet clear. METHODS: Thirty patients (average age 9.9 years) with obstructive lesions secondary to Kawasaki disease underwent repeated coronary artery angiography and thallium myocardial scintigraphy over a mean period of 7.7 years after the acute onset of Kawasaki disease. RESULTS: In the 27 patients who were enrolled in the acute phase of the disease because of coronary artery aneurysms, the later transformation to obstructive lesions was not significantly different between the 61 large and 6 medium-sized aneurysms. Obstructive transformation of aneurysms was more rapid in the right than in the left coronary artery (p < 0.001). From the last coronary angiogram obtained, the obstructive lesions were classified as localized stenosis > 90% in 10 vessels, occlusions in 6 vessels and segmental stenosis in 26 vessels. Both localized and segmental stenosis occurred significantly more often in the left anterior descending and the right coronary artery than in other vessels (p < 0.05). The incidence of collateral vessels was significantly correlated with a younger age at onset of Kawasaki disease, especially in patients with segmental stenosis (p < 0.001). Collateral vessels did not develop in the presence of localized stenosis regardless of the occurrence of myocardial ischemia. All occluded vessels had collateral development regardless of the presence of myocardial infarction. CONCLUSIONS: The treatment of localized stenosis may play an important role in preventing myocardial infarction in the chronic phase of Kawasaki disease. PMID- 8417059 TI - Coronary sinus sampling of cytokines after heart transplantation: evidence for macrophage activation and interleukin-4 production within the graft. AB - OBJECTIVES: This study was undertaken to evaluate the organ-specific release of cytokines after heart transplantation and to assess any correlation with transplant rejection. This cytokine profile should document the relative activation of mononuclear cell subsets within the graft. BACKGROUND: Up to 60% of mononuclear cells infiltrating the cardiac allograft during rejection are macrophages, but their role is undetermined. The T lymphocytes are activated, but the activity of specific T cell subsets is not known. We sought to assess for the first time in humans the in vivo activation of mononuclear cell subsets by measuring coronary sinus cytokine levels after heart transplantation. METHODS: Paired superior vena cava and coronary sinus serum samples were assayed for interleukin (IL)-2, IL-4 and IL-6, soluble IL-2 receptors, tumor necrosis factor alpha and neopterin in 10 patients at the time of 40 routine endomyocardial biopsy procedures. All cytokine measurements were made by using enzyme-linked immunosorbent assay; neopterin was measured by using radioimmunoassay. RESULTS: Interleukin-2 levels were not detectable (< 0.8 U/ml) in either the superior vena cava or the coronary sinus in the presence or absence of rejection. Interleukin-2 receptor levels were uniformly elevated to 1,283 U/ml in the superior vena cava and to 1,232 U/ml in the coronary sinus, with no correlation with rejection severity. Interleukin-4 levels were consistently higher in coronary sinus serum than in peripheral blood (229 vs. 61 pg/ml, p < 0.0005), but there was no relation with rejection. Interleukin-6 levels were higher in the coronary sinus than in the superior vena cava (200 vs. 120 pg/ml, p < 0.05). Tumor necrosis factor-alpha showed consistently elevated levels in coronary sinus serum (68 vs. 17 pg/ml, p < 0.0005), with no relation with rejection. Neopterin, which is produced only by activated macrophages, was also consistently elevated in the coronary sinus (2.5 vs. 2.2 nmol, p = 0.08). CONCLUSIONS: The cardiac allograft is a major source of cytokines after heart transplantation. The cytokine profile allows the activity of subsets of the mononuclear cell infiltrate to be investigated. Elevated coronary sinus activity of the macrophage-specific metabolite neopterin suggests macrophage activation within the allograft. This possibility is supported by elevated coronary sinus levels of tumor necrosis factor-alpha and IL-6. The T lymphocytes are activated, as evidenced by high soluble IL-2 receptor levels, but IL-2 production was suppressed by conventional immunosuppressive therapy. Coronary sinus IL-4 levels represent T helper-2 cell activation within the graft despite immunosuppression. We could find no temporal relation between the coronary sinus or superior vena cava cytokine concentration or profile and severity of rejection on concurrent biopsy studies. PMID- 8417060 TI - Failure of calcium channel blockers to improve ventricular relaxation in humans. AB - OBJECTIVES: The objective of this study was to ascertain whether the reversal of low peak filling rates after administration of calcium channel blockers in patients with diastolic dysfunction indicates true improvement in the rate of ventricular relaxation and left ventricular end-diastolic pressure measured by invasive indexes. BACKGROUND: Depressed filling rates measured noninvasively have been associated with diastolic dysfunction, specifically abnormal relaxation of the left ventricle. There is a reversal of these low peak filling rates after administration of calcium channel blockers. METHODS: Doppler echocardiographic measurements of peak filling rates were made and invasive high fidelity manometer tipped pressures were measured before and after administration of verapamil (0.1 mg/kg body weight) in 20 patients with coronary artery disease who had an ejection fraction > 40% and decreased peak filling rates. RESULTS: Verapamil caused significant increases in the peak filling rate, as measured by early transmitral (E) flow velocity, from 0.57 +/- 0.16 m/s to 0.77 +/- 0.15 m/s (p < 0.01), indicating reversal of decreased peak filling rates. Concomitantly, left ventricular end-diastolic pressure increased from 18.0 +/- 7.7 mm Hg to 24.1 +/- 9.0 mm Hg (p < 0.001). The time constant of relaxation was variable, with an overall significant increase from 45.8 +/- 10.4 ms to 53.2 +/- 14.6 ms (p = 0.01). CONCLUSIONS: Verapamil administered intravenously produced reversal of decreased peak filling rates in patients with coronary artery disease and normal ventricular function. However, there was an increase in left ventricular end diastolic pressure as well as an overall prolongation of the time constant of relaxation. Therefore, changes in peak filling rates do not accurately reflect the response of ventricular relaxation to drug interactions. Thus, calcium channel blockers should be used cautiously in the empiric treatment of patients with diastolic dysfunction. PMID- 8417061 TI - Intracardiac echocardiography in humans using a small-sized (6F), low frequency (12.5 MHz) ultrasound catheter. Methods, imaging planes and clinical experience. AB - OBJECTIVES: This study was designed to determine the clinical utility and feasibility of using 12.5-MHz ultrasound catheters for intracardiac echocardiography. BACKGROUND: Intracardiac echocardiography is a potentially useful technique of cardiac imaging and monitoring in certain settings. The feasibility of intracardiac echocardiography using 20-MHz ultrasound catheters in patients has been demonstrated. High resolution images of normal cardiac structures as well as cardiac abnormalities have been obtained. However, imaging has been limited by the shallow depth of field inherent in high frequency ultrasound imaging. METHODS: Intracardiac echocardiography with 12.5-MHz catheters was performed in eight mongrel dogs and 92 patients. Catheters were introduced percutaneously in 80 patients studied in the catheterization laboratory and directly into the heart in 12 patients in the operating room. Right heart imaging was performed in 68 patients and arterial and left heart imaging in 35 patients. RESULTS: When these catheters were introduced into the venous system, the right atrium, tricuspid valve, right ventricle, pulmonary valve and pulmonary artery were visualized. Pericardial effusion, intracardiac masses and atrial septal defects were correctly identified. The left ventricle, left atrium, mitral valve, aortic valve, aorta and coronary arteries could be imaged from the arterial circulation. Diseases identified included valvular aortic stenosis, subvalvular aortic stenosis and Kawasaki disease. Average imaging time was 10 min. No complications occurred as a result of intracardiac echocardiography. CONCLUSIONS: Intracardiac echocardiography with 12.5-MHz ultrasound catheters is safe and feasible; it also provides anatomic and physiologic information. This feasibility study provides a foundation for wider clinical use of intracardic echocardiography. PMID- 8417062 TI - Increased echodensity of transiently asynergic myocardium in humans: a novel echocardiographic sign of myocardial ischemia. AB - OBJECTIVES: This study was conducted to establish whether changes in myocardial texture can be observed in humans by transthoracic echocardiography during ischemic episodes of different severity and duration induced by various pathogenetic mechanisms. BACKGROUND: Increased echo-reflectivity of ischemic myocardium has been detected in experimental animals by epicardial echocardiography and by backscatter evaluation. METHODS: Transthoracic two dimensional echocardiographic monitoring with a commercially available electronic sector scanner (2.25- or 3.5-MHz transducer) was performed during 35 episodes of transient myocardial ischemia induced by ergonovine in patients with vasospastic angina (n = 9), by dipyridamole in patients with angiographically assessed coronary artery disease (n = 11) and by balloon occlusion during coronary angioplasty (n = 15). Quantitative texture analysis of gray levels was performed off-line on digitized images during rest conditions, ischemia and the recovery phase in regions showing normal contraction at rest, obvious dyssynergy during ischemia and normal contraction in the recovery phase. In each condition, a control region with normal contraction throughout the study was also evaluated. RESULTS: Chest pain occurred in 23 of the 35 episodes; electrocardiographic (ECG) changes were present in 26 episodes, and consisted of ST segment elevation in 13, ST segment depression in 10 and pseudonormalization of a basally negative T wave in 3. The duration of ischemic episodes was 67 +/- 53 s by symptomatic criteria and 91 +/- 52 s by ECG criteria. The risk region showed an increased end diastolic mean gray level amplitude in a.u. (arbitrary units) during ischemia (57 +/- 19) compared with rest (38 +/- 15) and recovery (38 +/- 18, p < 0.01). No significant changes were detected in the control region (rest 36 +/- 16 vs. ischemia 34 +/- 18 vs. recovery 31 +/- 13, p = NS). The percent increase in mean gray level was similar in the various types of stress employed (ergonovine, dipyridamole or angioplasty) and was not significantly correlated with either the duration of ST segment shift (r = 0.05, p = NS) or the severity of dyssynergy evaluated semiquantitatively by means of the wall motion score (r = 0.28, p = NS). In the 15 balloon occlusions performed in six patients during coronary angioplasty, the increased echoreflectivity of the risk zone was already evident during echocardiographic sampling performed after 10 +/- 4 s of occlusion (rest 35 +/- 9 vs. 53 +/- 10 a.u., p < 0.01) when no dyssynergy could be detected by quantitative wall motion analysis (percent area change by fixed center of mass reference system 31 +/- 10% at rest vs. 32 +/- 11% after 10 s of occlusion, p = NS). CONCLUSIONS: Transient short-lasting myocardial ischemia is associated with an abrupt increase in myocardial echodensity detectable by videodensitometric analysis applied to standard transthoracic echocardiographic images and is largely independent of the underlying pathogenetic mechanism (reduced blood supply or flow maldistribution with coronary stenosis). During controlled coronary occlusion, increased echodensity precedes the onset of regional dyssynergy. PMID- 8417063 TI - Usefulness of transesophageal echocardiography in identifying small left ventricular apical thrombus. AB - OBJECTIVES: The aim of this study was to determine whether transesophageal echocardiography could clarify the nature of equivocal echodense structures in the left ventricular apical region frequently found on transthoracic echocardiography by directing the ultrasound beam from the left ventricular base to the apex and achieving better image quality. BACKGROUND: Transthoracic echocardiography often reveals an echogenic structure suggesting thrombus in the left ventricular apical region because of limited near-field resolution and echo vibration artifact in apical views. METHODS: Thirty-six patients with coronary artery disease or dilated cardiomyopathy who had apical wall motion abnormalities and equivocal transthoracic echodense structures were studied with transesophageal echocardiography using special manipulation of the transesophageal probe for adequate imaging of the apical region. Left ventricular thrombus was defined when echogenic structures with a clearly delineated margin adjacent to but distinct from the endocardium were observed in at least two different tomographic views in the four-chamber and left ventricular long-axis views during both systole and diastole. RESULTS: Left ventricular thrombus (mean size 1.3 +/- 0.7 cm2) was defined by transesophageal echocardiography in 19 (53%) of 36 patients with suspected thrombus on transthoracic echocardiography in the four-chamber or left ventricular long-axis view. Heavy trabeculation or extremely high echo reflection, or both, was observed in the apical region in 12 patients (33%). No extra structures in the apical region were found in five patients. In 19 patients with transesophageal echocardiographically defined thrombus, 6 patients (31%) experienced arterial embolic events before the transesophageal procedure. In contrast, none of 17 patients without transesophageal echocardiographically defined thrombi had systemic embolism (p < 0.03). CONCLUSIONS: 1) Transesophageal echocardiography is useful in identifying left ventricular apical thrombus in patients with unclear echogenic structures on transthoracic apical images; and 2) the high incidence of arterial embolism in patients with transesophageal echocardiographically detected left ventricular thrombus indicates the clinical importance of such thrombus. PMID- 8417064 TI - Implication of negative results on a monoplane transesophageal echocardiographic study in patients with suspected infective endocarditis. AB - OBJECTIVES: This study was conducted to determine the implications of negative findings on a transesophageal echocardiographic study in which neither a vegetation nor an abscess is identified in patients with clinically suspected infective endocarditis. BACKGROUND: Echocardiography is the procedure of choice for evaluating suspected infective endocarditis in patients. Transesophageal echocardiography has been shown to be superior to transthoracic imaging. Although the importance of positive results or a diagnostic study is known, the significance of negative findings on a transesophageal study is not clear. METHODS: All transesophageal echocardiographic studies performed over a 2-year period for suspected infective endocarditis were reviewed and the clinical course of patients with an initially negative study result was assessed to determine their final diagnosis. RESULTS: Of the 105 patients identified, 65 had a negative transesophageal study result. In the majority of this group (56 of 65), an alternate diagnosis was made or there was no infective endocarditis on follow-up examination, or both. Of the remaining nine patients, four were treated for endocarditis without a definite diagnosis and five had infective endocarditis proved by either repeat transesophageal study (n = 3), pathologic findings (n = 1) or a diagnostic clinical course (n = 1). Gram-positive bacteremia and the presence of a prosthetic valve in the aortic position tended to be more common in the latter group. CONCLUSIONS: A negative transesophageal study result reduces the likelihood that endocarditis is present. Repeat examination, however, should be considered in high risk patients, such as those with prosthetic valves or unexplained bacteremia, to avoid a missed diagnosis. PMID- 8417065 TI - Suspected bacterial endocarditis: to TEE or not to TEE. PMID- 8417066 TI - Passive smoking increases experimental atherosclerosis in cholesterol-fed rabbits. AB - OBJECTIVES: We evaluated the influence of passive smoking on experimental atherosclerosis in cholesterol-fed rabbits. BACKGROUND: Exposure to environmental tobacco smoke (ETS) has been epidemiologically linked to death from ischemic heart disease in nonsmokers. METHODS: New Zealand male rabbits were randomly divided into three groups after 2 weeks of a 0.3% cholesterol diet. Sixteen rabbits were exposed to a high and 16 rabbits to a low dose of ETS; 32 rabbits located in another room served as an unexposed control group. After 10 weeks of ETS exposure, all rabbits were killed, and the percent of aortic and pulmonary artery endothelial surfaces covered by lipid lesions was measured by staining and planimetry. RESULTS: Average air nicotine, carbon monoxide and total particulate concentrations were 1,040 micrograms/m3, 60.2 ppm and 32.8 mg/m3 for the high dose ETS group, 30 micrograms/m3, 18.8 ppm and 4.0 mg/m3 for the low dose ETS group and < 1 microgram/m3, 3.1 ppm and 0.13 mg/m3 for the control group. The percent atherosclerotic involvement of the aorta and pulmonary artery increased significantly with ETS exposure (for the aorta, 30 +/- 19% [mean +/- SD] for the control group, 36 +/- 14% for the low dose ETS group and 52 +/- 21% for the high dose ETS group, p < 0.001; for the pulmonary artery, 22 +/- 15% for the control group, 29 +/- 25% for the low dose ETS group, and 45 +/- 12% for the high dose ETS group, p < 0.001). Bleeding time was significantly shorter in the two ETS groups than in the control group (86 +/- 17 vs. 68 +/- 15, 68 +/- 18 s, p < 0.001). There were no significant differences in serum triglycerides, cholesterol and high density lipoprotein cholesterol at the end of the study. CONCLUSIONS: Environmental tobacco smoke affects platelet function and increases aortic and pulmonary artery atherosclerosis. This increase of atherosclerosis was independent of changes in serum lipids and exhibited a dose-response relation. These results are consistent with data from epidemiologic studies demonstrating that ETS increases the risk of death due to heart disease. PMID- 8417067 TI - Role of sodium/calcium exchange in the mechanism of myocardial stunning: protective effect of reperfusion with high sodium solution. AB - OBJECTIVES: This study was conducted to elucidate the role of sodium/calcium (Na+/Ca2+) exchange in the mechanism of myocardial stunning. BACKGROUND: Cellular Ca2+ overload mediated by Na+/Ca2+ exchange during reperfusion has been proposed as a mechanism for myocardial stunning. Because no specific pharmacologic inhibitors of the exchanger are available, we increased extracellular sodium concentration ([Na]o) during the early phase of reperfusion to decrease the driving force for Ca2+ influx through the pathway. METHODS: Isovolumetric left ventricular pressure and phosphorus-31 nuclear magnetic resonance spectra were measured in isolated perfused ferret hearts. Hearts were reperfused with different solutions after 15 min of total global ischemia at 37 degrees C. RESULTS: Hearts reperfused with standard solution ([Na]o = 140 mmol/liter; the stunned hearts, n = 8) showed only 69 +/- 3% (mean +/- SEM) recovery of developed pressure relative to preischemic control developed pressure. In contrast, hearts reperfused with a high [Na]o solution ([Na]o = 268 mmol/liter) during the initial 5 min, followed by a gradual decrease of [Na]o to the standard level over 25 min (the high [Na]o group, n = 8) showed significantly better recovery of developed pressure (85 +/- 2%, p < 0.05 vs. the stunned hearts). In contrast, reperfusion with solutions in which the additional Na was substituted either by 256 mmol/liter sucrose or 128 mmol/liter choline chloride did not improve functional recovery, indicating that the beneficial effects of high [Na]o reperfusion are not due to either high ionic strength or high osmolarity. Phosphorus-31 nuclear magnetic resonance spectra showed no correlation between functional recovery and intramyocardial contents of phosphorus compounds or pH. CONCLUSIONS: High [Na]o reperfusion protects against stunning, supporting the concept that Na+/Ca2+ exchange plays an important role in the mechanism of stunned myocardium. PMID- 8417068 TI - Quantitative analysis of elastic recoil after balloon angioplasty and after intracoronary implantation of balloon-expandable Palmaz-Schatz stents. AB - OBJECTIVES: The purpose of this study was to measure elastic recoil from sequential angiograms after balloon angioplasty and after implantation of a balloon-expandable Palmaz-Schatz stent in the same patient, and to compare the results with the late angiographic outcome. BACKGROUND: The immediate result of coronary balloon angioplasty is influenced by plastic deformation, primarily of the atherosclerotic plaque, and by elastic recoil, primarily of the less or nondiseased vessel circumference. METHODS: The extent of elastic recoil was measured quantitatively as the difference between maximal balloon size and the resulting vessel diameter or cross-sectional area. RESULTS: Analysis was performed in 60 patients who received a single stent for late restenosis after initially successful coronary balloon angioplasty. Minimal lumen diameter (minimal cross-sectional area) was 0.98 +/- 0.43 mm (0.97 +/- 0.67 mm2) before balloon angioplasty, 2.06 +/- 0.36 mm (3.68 +/- 1.17 mm2) after angioplasty (both p < 0.001 vs. values before angioplasty) and 2.98 +/- 0.26 mm (7.12 +/- 1.28 mm2) after stenting (both p < 0.001 vs. postangioplasty results). No significant changes in vessel reference diameters or areas were measured. Mean balloon/artery ratios were similar in both procedures, ranging from 0.93 to 0.96. The calculated mean elastic recoil was 0.98 +/- 0.50 mm in diameter (31%) and 3.67 +/- 2.05 mm2 in area (48%) after balloon angioplasty compared with 0.10 +/- 0.07 mm (3.5%) and 0.38 +/- 0.36 mm2 (5.1%) after stenting. Increasing balloon sizes induced increased vessel stretch, which was followed by increased elastic recoil in the angioplasty group in contrast to the stenting group. Short, noncalcified and eccentric lesions tend to be associated with increased recoil after balloon angioplasty. Overdilation or underdilation in one of the procedures, changes in postprocedural vasomotion or postprocedural thrombus formation was not responsible for this outcome. After 6 months mean minimal lumen diameter was 2.39 +/- 0.58 mm, suggesting a mean hyperplasia of 0.59 +/- 0.51 mm. Twelve patients (20%) had a follow-up diameter that was equal to or less than the mean postangioplasty result and eight patients (14%) had a diameter stenosis of > 50%. CONCLUSIONS: The implantation of a Palmaz-Schatz stent almost completely eliminates the decrease in vessel dimensions caused by elastic recoil and therefore diminishes the impact of hyperplasia and reduces the rate of restenosis. PMID- 8417069 TI - Transient time course of cocaine-induced cardiac depression versus sustained peripheral vasoconstriction. AB - OBJECTIVES: The goal of this study was to define in vivo the magnitude and temporal course of cocaine-induced cardiac depression separate from peripheral vascular loading changes. BACKGROUND: Cocaine administration immediately alters arterial pressure, ventricular filling, contractility and heart rate. These combined changes have complicated previous analyses of the cardiac effects of cocaine, because routine measures of function are influenced by all these factors. METHODS: Pressure-volume loops and relations were measured by conductance catheter technique in 10 intact chest anesthetized dogs before and up to 30 min after administration of cocaine (3 mg/kg intravenously). Heart rate was kept constant by atrial pacing, and data were obtained before and after ganglionic blockade. RESULTS: Cocaine decreased ejection fraction, cardiac output and maximal rate of rise of left ventricular pressure (dP/dtmax) and increased arterial vascular resistance (all p < 0.05). The maximal change occurred by 2 min and was fully sustained for the next 30 min. In contrast, contractile indexes based on pressure-volume analysis (e.g., end-systolic pressure-volume relation) decreased only briefly at 2 min (by -19%) and returned to control value after 5 to 10 min. Reflex blockade with hexamethonium eliminated cocaine-induced vasoconstriction, but the magnitude and brevity of cardiac depression were unaltered. Ejection fraction, dP/dtmax and cardiac output now also decreased transiently, suggesting that the sustained changes observed before blockade in these variables were load related. Analogous load effects also explained changes in peak filling rate. CONCLUSIONS: Cocaine induces brief direct cardiac depression that is temporally dissociated from more sustained peripheral vasoconstriction. This dissociation is not measured by traditional left ventricular function analysis because of simultaneous load change. The transience of cardiac depression suggests that it probably does not play a major role in late adverse sequelae of cocaine administration. PMID- 8417070 TI - Use of nonionic or low osmolar contrast agents in cardiovascular procedures. American College of Cardiology Cardiovascular Imaging Committee. AB - Low osmolar contrast agents produce less adverse electrophysiologic and hemodynamic alterations during cardiac catheterization. The nonionic agents probably reduce the risk of provoking myocardial ischemia during coronary arteriography or ventriculography. Patients also report less subjective sensation of discomfort during administration of low osmolar agents for cardiovascular procedures. However, nonionic agents have not been proved to reduce the incidence of several serious complications of cardiac catheterization, including acute renal failure and anaphylactoid reaction. Although evidence is inconclusive, there may be an increased risk of thromboembolic complications during cardiac catheterization when certain low osmolar nonionic agents are administered. Nonionic contrast agents have not been definitely proved to reduce the risk of death after cardiac catheterization. PMID- 8417071 TI - Which references should be cited? PMID- 8417072 TI - Access to cardiovascular care. ACC policy statement. American College of Cardiology. PMID- 8417073 TI - Ventricular arrhythmia and silent myocardial ischemia. PMID- 8417074 TI - Intracoronary ultrasound imaging: correlation of plaque morphology with angiography, clinical syndrome and procedural results in patients undergoing coronary angioplasty. AB - OBJECTIVES: This study was designed to establish the relation between ultrasound derived atheroma morphology and the clinical, procedural and angiographic features of patients presenting for coronary angioplasty. BACKGROUND: Intracoronary ultrasound imaging provides accurate dimensional information regarding arterial lumen and wall structures. Atheroma composition may also be assessed by ultrasound; however, only limited studies have been performed in patients. METHODS: In 65 patients a diagnostic ultrasound imaging catheter or a combination imaging-angioplasty balloon catheter was used during coronary angioplasty to image both the lesion and the vessel segment just proximal to it (reference segment). Ultrasound images were analyzed for lumen, total vessel and plaque areas and were classified into five morphologic subtypes (soft, fibrous, calcific, mixed plaque and concentric subintimal thickening). These data were compared with angiographic morphologic features, procedural results and clinical angina pattern (stable vs. unstable). RESULTS: Morphologic analysis of the ultrasound images obtained from the lesion correlated well with the clinical angina syndrome. Compared with patients with stable angina, patients with unstable angina had more soft lesions (74% vs. 41%), fewer calcified and mixed plaques (fibrotic, soft or calcific components in one or more combinations [25% vs. 59%]) and fewer intralesional calcium deposits (16% vs. 45%) (all p < 0.01). There was no correlation between ultrasound and angiographic lesion morphologic characteristics for either the reference segment or the lesion. Ultrasound demonstrated greater sensitivity than angiography for identifying unstable lesions (74% vs. 40%). Dimensional analysis demonstrated a large plaque burden in the reference segments (45 +/- 15% of total vessel area). Postangioplasty plaque burden was also high (62 +/- 9%). There was a significant, but only fair correlation between lumen area determined by angiography and ultrasound for both the reference segment (r = 0.70, p < 0.001) and the postangioplasty lesion (r = 0.63, p < 0.05). CONCLUSIONS: Morphologic plaque classification by ultrasound is closely correlated to clinical angina but has little relation to established angiographic morphologic characteristics. Intracoronary ultrasound imaging during angioplasty identifies a large residual plaque burden in both the reference segment and the lesion. In the future, determination of plaque composition by intracoronary ultrasound may be important in selecting or modifying interventional therapeutic options. PMID- 8417075 TI - Histology after stenting of human saphenous vein bypass grafts: observations from surgically excised grafts 3 to 320 days after stent implantation. AB - OBJECTIVES: To gain insight into the mechanism of stenting in humans and its short- and long-term implications, we studied the vascular wall of saphenous vein aortocoronary bypass grafts after implantation of the Wallstent. BACKGROUND: The implantation of a stent in aortocoronary bypass grafts may provide an alternative solution for revascularization in patients who are poor candidates for reoperation. Because human histopathologic findings after stenting with the Wallstent have not previously been described in detail, we examined graft segments that were surgically retrieved from 10 patients (21 stents) at 3 days to 10 months after implantation of the stent. METHODS: The grafts were examined by a combination of the following techniques: light microscopy, immunocytochemistry and both scanning and transmission electron microscopy. RESULTS: Early observations revealed that large amounts of platelets and leukocytes adhered to the stent wires during the first few days. At 3 months, the wires were embedded in a layered new intimal thickening, consisting of smooth muscle cells in a collagenous matrix. In addition, foam cells were abundant near the wires. Extracellular lipids and cholesterol crystals were found after 6 months. Smooth muscle cells and extracellular matrix formed the predominant component of restenosis. This new intimal thickening was lined with endothelium, in some cases showing defect intercellular junctions and abnormal adherence of leukocytes and platelets as late as 10 months after implantation. CONCLUSIONS: This type of stent is potentially thrombogenic and seems to be associated with extracellular lipid accumulation in venous aortocoronary bypass grafts. PMID- 8417076 TI - Angiographic validation of bedside markers of reperfusion. AB - OBJECTIVES: We sought to validate with coronary angiography several primary and ancillary markers of reperfusion. BACKGROUND: The availability of bedside markers of reperfusion is of major importance in the thrombolytic therapy of acute myocardial infarction. However, the reliability of current markers is still controversial. METHODS: Changes in chest pain, ST segment elevation and heart rate and rhythm were assessed every 5 to 10 min for up to 3 h after initiation of recombinant tissue-type plasminogen activator therapy in 82 patients with acute myocardial infarction. Coronary angiography was performed within 24 h. RESULTS: At angiography, 69 of the 82 patients had a patent infarct-related artery with Thrombolysis in Myocardial Infarction trial (TIMI) grade 3 flow and a rapid and progressive decrease in chest pain and ST elevation. The pain resolved in 24 +/- 23 min (range 3 to 50). The ST elevation decreased by > or = 50% within 16 +/- 14 min (range 5 to 41). Accelerated idioventricular rhythm developed in 49% of patients and sinus bradycardia in 23%; conduction abnormalities and atrial fibrillation resolved. All markers appeared in close temporal proximity to the onset of an abrupt increase in plasma creatine kinase (CK) and CK-MB isoenzyme activity, a previously validated marker of the time of reperfusion. Before its final resolution, ST elevation transiently decreased and increased in 58% of patients. Comparison of one pretreatment and one posttreatment electrocardiogram significantly reduced the reliability of ST segment change as a marker of reperfusion. In 13 of 82 patients, the infarct-related artery demonstrated TIMI grade < or = 2 flow; in 9, pain and ST elevation did not lessen and CK and CK-MB activity showed no abrupt increase. The remaining four patients initially demonstrated a decrease in pain and ST elevation; however, within 3 h and before angiography, the recurrence of pain and ST elevation suggested reocclusion. CONCLUSIONS: A rapid and progressive decrease in pain and ST elevation is a reliable marker of reperfusion with TIMI grade 3 flow. Because ST elevation and pain often fluctuate before undergoing final resolution with reperfusion, frequent or continuous monitoring of ST elevation is essential for reliable recognition of the fact and time of reperfusion. Accelerated idioventricular rhythm and episodes of sudden sinus bradycardia, although specific to reperfusion, do not occur in all patients with reperfusion. PMID- 8417077 TI - Can restenosis after coronary angioplasty be predicted from clinical variables? AB - OBJECTIVES: The purpose of this study was to determine whether variables shown to correlate with restenosis in one group (learning group) could be shown to predict recurrent stenosis in a second group (validation group). BACKGROUND: Restenosis remains a critical limitation after percutaneous transluminal coronary angioplasty. Although several clinical variables have been shown to correlate with restenosis, there are few data concerning attempts to predict recurrent stenosis. METHODS: The source of data was the clinical data base at Emory University. Patients who had had previous coronary surgery and patients who underwent coronary angioplasty in the setting of acute myocardial infarction were excluded. A total of 4,006 patients with angiographic restudy after successful angioplasty were identified. They were classified into a learning group of 2,500 patients and a validation group of 1,506 patients. The correlates of restenosis in the learning group were determined by stepwise logistic regression, and a model was developed to predict the probability of restenosis and was tested in the validation group. By using various cut points for the predicted probability of restenosis, a receiver operating characteristic curve was created. Goodness of fit of the model was evaluated by comparing average predicted probabilities with average observed probabilities within subgroups on the basis of risk level determined by linear regression analysis. RESULTS: In the learning group 1,145 patients had restenosis and 1,355 did not. Correlates of restenosis were severe angina, severe diameter stenosis before angioplasty, left anterior descending coronary artery dilation, diabetes, greater diameter stenosis after angioplasty, hypertension, absence of an intimal tear, eccentric morphology and older patient age. The model derived from the learning group was used to predict restenosis in the validation group. By varying the cut point for the predicted probability of restenosis above which restenosis is diagnosed and below which it is not, a receiver operating characteristic curve was created. The curve was close to the line of identity, reflecting a poor predictive ability. However, the model was shown to fit well with the predicted probability of restenosis correlating well with the observed probability (r = 0.98, p = 0.0001). CONCLUSIONS: Clinical variables provide limited ability to predict definitively whether a particular patient will have restenosis. However, the current model may be used to predict the probability of restenosis, with some uncertainty, at least in well characterized patients who have already had angioplasty. PMID- 8417078 TI - Is ST segment re-elevation associated with reperfusion an indicator of marked myocardial damage after thrombolysis? AB - OBJECTIVES: The significance of ST segment re-elevation at reperfusion by thrombolysis was evaluated. BACKGROUND: The significance of ST re-elevation has not been studied. Hence, we evaluated whether ST re-elevation is an indicator of marked myocardial necrosis after reperfusion. METHODS: Twelve-lead electrocardiograms were recorded serially, before thrombolysis and immediately after each coronary angiographic procedure during thrombolysis. RESULTS: In 32 patients with acute myocardial infarction, 15 showed transient ST re-elevation at reperfusion (group 1) and 17 showed reduction (group 2). Peak creatine kinase (CK) and CK-MB isoenzyme activity levels were significantly higher in group 1 than in group 2. Twelve patients in group 1 had strongly positive findings on early technetium-99m pyrophosphate scintigraphy, compared with one patient in group 2 (p < 0.001). The regional ejection fraction did not increase from the acute phase to the chronic phase in group 1. The ST deviation before thrombolysis was significantly greater in group 1 than in group 2 (p < 0.001). All 14 patients in group 1 showed Thrombolysis in Myocardial Infarction (TIMI) grade 0 flow and 12 of these patients did not have good collateral flow before thrombolysis. CONCLUSIONS: These data suggest that 1) ST re-elevation at reperfusion is a sign of limited myocardial salvage by thrombolysis, and 2) high ST elevation and TIMI grade 0 flow without good collateral flow before thrombolysis may be predictive variables for marked myocardial necrosis after reperfusion. PMID- 8417079 TI - A new method for assessment of collateral development after acute myocardial infarction. AB - OBJECTIVES: The purpose of this study was to test the hypothesis that the diameter of the recipient coronary artery of the well developed collateral circulation in patients with acute myocardial infarction increases because of the augmented intravascular pressure caused by subsequent collateral development. BACKGROUND: It is well known that collateral circulation develops after acute myocardial infarction. However, some patients have a well developed collateral circulation at the onset of infarction, which may limit the angiographic evaluation of further development of collateral circulation. METHODS: We measured the diameter of the donor and recipient arteries of the collateral circulation by means of a computer-assisted analysis system in seven patients with acute myocardial infarction who had a totally occluded infarct-related coronary artery during the acute and chronic stages of infarction. All coronary angiograms were obtained after the administration of sublingual nitroglycerin. The measurement was repeated immediately after (within 6 h) and late after (42 +/- 11 days) the onset of acute myocardial infarction. RESULTS: The diameter of the donor artery remained unchanged (1.32 +/- 0.98 vs. 1.42 +/- 1.12 mm). In contrast, the diameter of the recipient artery increased from 1.25 +/- 0.63 to 1.55 +/- 0.61 mm (p < 0.01). These changes in coronary artery diameter were associated with an improvement in regional myocardial wall motion at rest in infarct areas (6.7 +/- 7.0% vs. 13.6 +/- 10.7%, p < 0.05). CONCLUSIONS: These findings indicate that serial measurement of coronary artery diameter is useful for the evaluation of collateral development after acute myocardial infarction. PMID- 8417080 TI - A unified functional/anatomic substrate for circus movement atrial flutter: activation and refractory patterns in the canine right atrial enlargement model. AB - OBJECTIVES: This study was designed to test the concept of a functional/anatomic interaction in a canine model of reentry based on right atrial enlargement and to elucidate the electrophysiologic basis for functional conduction block. BACKGROUND: The monotonic feature of atrial flutter suggests a uniform substrate for the arrhythmia. Atrial flutter in the sterile pericarditis model is due to single-loop circus movement around a functional or a functional/anatomic obstacle near the atrioventricular (AV) ring. Sustained circus movement requires a critical interaction of a functional arc of block, a natural obstacle, the AV ring and a zone of slow conduction. The location of the inferior vena cava predisposes the lower right atrium to single-loop reentry. METHODS: In 11 dogs with right atrial enlargement, 127 bipolar epicardial electrograms were obtained during atrial flutter. For correlation of activation and refractory maps, the effective refractory period under each electrode was determined using the extrastimulus technique. RESULTS: Atrial flutter was due to single-loop reentry around functional arcs of block near the AV ring (n = 2) or around functional/anatomic obstacles (n = 8) involving the inferior vena cava. A slow zone was located between the arc and the AV ring and between the inferior vena cava and AV ring, respectively. During initiation, the arc joined the AV ring, forcing activation to proceed around the free end of the arc before breaking through the arc near the AV ring. Arrhythmia termination required the arc of block to rejoin the AV ring. Inducibility of sustained atrial flutter was associated with a marked spatial dispersion of refractoriness. The configuration of the functional arc of block was critically dependent on the spatial pattern of refractoriness. CONCLUSIONS: Atrial flutter requires a similar functional or functional/anatomic substrate independent of the underlying etiology. The spatial distribution of refractoriness in enlarged canine atria provides an adequate substrate for the development of functional conduction block. PMID- 8417081 TI - Differentiation of paroxysmal narrow QRS complex tachycardias using the 12-lead electrocardiogram. AB - OBJECTIVES: The purpose of this study was to evaluate the utility of the 12-lead electrocardiogram (ECG) for differentiating paroxysmal narrow QRS complex tachycardias. BACKGROUND: Previous studies evaluating the utility of the 12-lead ECG for differentiating paroxysmal supraventricular tachycardia types have shown conflicting results on the usefulness of some ECG criteria, and some criteria that are considered to be useful have never been formally evaluated. METHODS: Two hundred forty-two ECGs demonstrating paroxysmal narrow QRS complex (< 0.11 ms) tachycardia (rate > or = 120 beats/min) were analyzed. All ECGs were analyzed by an observer who had no knowledge of the mechanism of the tachycardia. RESULTS: There were 137 atrioventricular (AV) reciprocating tachycardias, 93 AV node reentrant tachycardias and 12 atrial tachycardias. Six criteria were found to be significantly different between tachycardia types by univariate analysis. A P wave separate from the QRS complex was observed more frequently in AV reciprocating tachycardia (68%) and atrial tachycardias (75%). A pseudo r' deflection in lead V1 and a pseudo S wave in the inferior leads were more common in AV node reentrant tachycardia (58% and 14%, respectively); QRS alternans was present more often during AV reciprocating tachycardia (27%). When a P wave was present, an RP/PR interval ratio > or = 1 was more common in atrial tachycardias (89%). During sinus rhythm, manifest pre-excitation was observed more often in patients with AV reciprocating tachycardia (45%). By multivariate analysis, the presence of a P wave separate from the QRS complex, pseudo r' deflection in lead V1, QRS alternans during tachycardia and the presence of pre-excitation during sinus rhythm were independent predictors of tachycardia type. These criteria correctly identified 86% of AV node reentrant tachycardias, 81% of AV reciprocating tachycardias and incorrectly assigned the tachycardia type in 19% of cases. CONCLUSIONS: Several features on the ECG are useful for differentiating supraventricular tachycardia type. However, approximately 20% of tachycardias may be incorrectly classified on the basis of analysis of the ECG; therefore, the ECG should not serve as the sole means for determining tachycardia mechanism. PMID- 8417082 TI - Polymorphic ventricular tachycardia induced by programmed stimulation: response to procainamide. AB - OBJECTIVES: This study was designed to evaluate the effects of procainamide on polymorphic ventricular tachycardia induced by programmed stimulation and to correlate the responses with heart disease, left ventricular endocardial activation abnormalities and the signal-averaged electrocardiogram (ECG). BACKGROUND: Polymorphic ventricular tachycardia is induced frequently during electrophysiologic studies. In many patients this response is an artifact of programmed stimulation; in others, it appears to be clinically relevant. Previous observations have suggested that in some patients type IA antiarrhythmic agents can change the response to programmed stimulation from polymorphic to uniform ventricular tachycardia. METHODS: Programmed right ventricular stimulation was performed in the absence of antiarrhythmic drugs and after procainamide. Signal averaged ECGs and left ventricular maps were performed during sinus rhythm in the absence of antiarrhythmic drugs. RESULTS: We evaluated 79 consecutive patients undergoing clinical electrophysiologic studies, in whom polymorphic ventricular tachycardia was the only arrhythmia induced in the absence of antiarrhythmic drugs. After procainamide administration, uniform monomorphic ventricular tachycardia was induced in 24 patients (Group 1), inducible polymorphic ventricular tachycardia persisted in 30 patients (Group 2) and no ventricular tachycardia could be induced in the remaining 25 patients (Group 3). Twenty-three (96%) of 24 patients developing uniform ventricular tachycardia after procainamide administration had coronary artery disease compared with 63% of Group 2 and 48% of Group 3 patients (p = 0.003). Left ventricular aneurysms were also found more frequently (46%) in the patients developing uniform ventricular tachycardia after procainamide than in either Group 2 or Group 3 (13% and 0%, respectively, p < 0.008). Abnormalities of the signal-averaged ECG typically seen in patients with spontaneous reentrant sustained ventricular tachycardia were significantly more frequent in patients who developed inducible uniform ventricular tachycardia after procainamide than in those who did not. Similarly, patients developing uniform ventricular tachycardia after procainamide had more extensive abnormalities of left ventricular endocardial activation revealed by catheter maps during sinus rhythm. CONCLUSIONS: The conversion of inducible polymorphic ventricular tachycardia to uniform ventricular tachycardia after procainamide administration occurs almost exclusively in patients with coronary disease, previous myocardial infarction and abnormal left ventricular function. This response may permit activation mapping of tachycardias, allowing the application of surgical or catheter ablation techniques that would otherwise not be possible in such patients. PMID- 8417083 TI - Clinical significance of polymorphic ventricular tachycardia induced by programmed stimulation. PMID- 8417084 TI - Enzyme replacement for patients with cystic fibrosis. PMID- 8417085 TI - Debating dietetics education: what do students need to succeed? PMID- 8417086 TI - Consumers confused by nutrition issues. PMID- 8417087 TI - America eats out: nutrition in the chain and family restaurant industry. PMID- 8417088 TI - Gender dominance in the work setting: implications for dietitians. PMID- 8417089 TI - Hospital-associated malnutrition: a reevaluation 12 years later. AB - A prospective evaluation of general medical patients at the University of Alabama at Birmingham was performed in 1976 and repeated in 1988 to determine change in malnutrition prevalence. Plasma folate, plasma ascorbate, weight for height, triceps skinfold, arm muscle circumference, lymphocyte count, albumin, and hematocrit measurements were combined to form a likelihood of malnutrition (LOM) score. The nutritional status of 228 consecutive patients was assessed by the LOM score at admission and at the 14th day of hospitalization and compared with 1976 findings. The same testing methods were used and the same patient diagnoses and demographic characteristics were found in 1988 and 1976. Of the patients staying more than 14 days, the length of stay was the same in 1988 and 1976 (30 days and 31 days, respectively). However, a smaller percentage of patients stayed 2 weeks or longer in 1988 (21% vs 33% in 1976). In 1988, high LOM scores at admission predicted longer lengths of stay and showed a trend toward increased mortality. The 1976 findings also showed that high LOM scores were associated with longer lengths of stay and increased mortality. LOM scores paired from admission to follow-up improved with stay in 1988 and worsened in 1976. The number of patients with high LOM scores at follow-up was lower in 1988 than in 1976 (46% and 62%, respectively). These findings indicate that identification of malnutrition indicators has improved since 1976. However, dietitians should continue to improve the nutrition assessment and intervention process. PMID- 8417090 TI - Guidelines vs practice in the delivery of diabetes nutrition care. AB - The American Dietetic Association and the American Diabetes Association have published recommendations for the nutrition care of people with diabetes. However, the frequency of this care is rarely documented. As part of a study of diabetes care and education practices, the Michigan Diabetes Research and Training Center collected extensive data from 440 randomly selected adults who receive diabetes care from community physicians. These data provided a basis for comparison between diabetes nutrition care as recommended and as delivered in typical American communities. In this population (mean age = 61 years; 54% women), 89% (393) had non-insulin-dependent diabetes mellitus (NIDDM). Of these, 152 were managed with insulin (NIDDM/I) and 241 were not managed with insulin (NIDDM/NI). Most of the NIDDM/NI group was overweight (71%) and had elevated levels of glycated hemoglobin (62%) and serum cholesterol (53%). Yet they were significantly less likely than those with NIDDM/I to see a dietitian. The most frequently reported reason for not seeing a dietitian was that a physician had not referred them (53%). More than 90% of those with NIDDM/I or NIDDM/NI who were referred to a dietitian saw one. Because this population was from randomly selected communities, physicians, and patients, the results are probably generalizable to other regions of the United States. This study shows that in community practice, insulin use is the primary marker of the need for nutrition intervention, and the lack of physician referral to a dietitian is an important barrier to people receiving recommended diabetes nutrition care. PMID- 8417091 TI - Foods perceived by adults as causing adverse reactions. AB - This study was undertaken to identify differences in offending foods reported by 45 patients with classic symptoms of food allergy and/or subjective food-related complaints not traditionally associated with food allergy. On the basis of a comprehensive clinical history, skin testing, and double-blind food challenges, patients were diagnosed as having confirmed (n = 22) or unconfirmed (n = 23) adverse reactions to foods. The majority of patients in both groups were women-17 of those with confirmed reactions and 21 of those with unconfirmed reactions. Individuals with confirmed reactions were significantly younger, 34.8 +/- 10.9 years, than those with unconfirmed reactions, 41.5 +/- 9.6 years. The symptoms and reaction patterns of adults with confirmed reactions were generally consistent with immunoglobulin E-mediated food hypersensitivity. Individuals with unconfirmed reactions were more likely to report frequently occurring, delayed onset symptoms of a nonspecific nature, and they had an average of five times as many foods causing adult-onset symptoms than those with confirmed reactions. Adults with confirmed reactions more often reported common food allergens such as tree nuts, legumes, and crustaceans; those with unconfirmed reactions were more apt to name foods not commonly implicated in adult food allergies confirmed by conventional diagnostic methods. The types of offending foods reported suggested that individuals with unconfirmed reactions were influenced by the popular news media and clinical ecology-oriented literature. Their use of nutrition supplements was consistent with the attitude of health activism. Such individuals may seek nontraditional health care or adopt questionable dietary practices. Individuals with adverse food reactions of a nonspecific nature pose challenging problems to dietitians. PMID- 8417092 TI - Body composition of patients with Alzheimer's disease. AB - Low body weight is frequently reported in patients with Alzheimer's disease. We sought to discover why by comparing the body composition of 28 cognitively normal elders and 23 institutionalized individuals with Alzheimer's disease. Body mass index was calculated from standing height and weight. Percentages of lean body mass, body fat, and body water were derived from bioimpedance measurements of resistance and reactance. Skinfold thickness was measured at seven body sites to estimate regional fat distribution. Variables were analyzed by analysis of variance with subjects grouped by cognitive status within gender. Activity level and age were not significant covariates. Both women and men with Alzheimer's disease weighed less than control subjects. Differences in body composition were more pronounced in women with Alzheimer's disease, who had lower body mass index (22.0 +/- 3.0 vs 26.1 +/- 5.1), higher percentage of lean body mass (73.8 +/- 5.1 vs 66.9 +/- 6.5), lower percentage of body fat (26.1 +/- 5.1 vs 33.1 +/- 6.5), and higher percentage of body water (55.8 +/- 5.0 vs 49.3 +/- 6.5) compared with control women. Except for lower body weight, the body composition of men with Alzheimer's disease was not significantly different from that of control men. Patients of both sexes with Alzheimer's disease had less truncal body fat compared with controls, which gave them a youthful body habitus. These differences were not accounted for by age, diet, or activity. Our findings indicate that patients with Alzheimer's disease have lower body weight and may require higher energy intake than cognitively normal elders. PMID- 8417093 TI - Comparison of determinants of frame size in older adults. AB - Accurate designations of body frame size can enhance the interpretation of height weight tables. Determinants of frame size should be quantifiable, reflective of skeletal dimensions, and not influenced by adiposity. Visual assessment, the height:wrist circumference ratio, elbow breadth by 1983 and 1984 standards, and Frame Index 2 were studied in 300 healthy adults over 64 years of age. Distribution of frame size across small, medium, and large categories revealed that visual assessment and height:wrist circumference ratio agreed with designations of elbow breadth measurements for less than 50% of the population. Highest levels of agreement occurred between elbow breadth (1984) and Frame Index 2. For men, high partial correlations between height:wrist circumference ratio and subscapular fatfold measurements corrected for age and arm muscle area indicated that measurement may be affected by body fat. Lowest correlations with subscapular fatfolds were for wrist and ankle breadths for women and ankle breadth for men. Elbow breadth and Frame Index 2 had low negative correlations with subscapular skinfold for men but much higher values for women. Elbow breadth measurements are widely used as frame size determinants, but for women, at least, ankle and wrist breadths meet the criterion of low associations with body fatness. PMID- 8417094 TI - Are n-3 fatty acids essential nutrients for fetal and infant development? AB - Recent research indicates that n-3 fatty acids (FAs) are essential nutrients in early human development. In human infants, nonhuman primates, and animal models, the n-3 FA, docosahexaenoic acid (DHA, 22:6n-3) is highly concentrated in brain and retinal tissues and accumulates during late fetal and early neonatal life. Diets deficient in n-3 FAs are associated with reduced levels of DHA in erythrocytes and brain and retinal tissues and with abnormalities in retinal function that may be irreversible. The precursor of DHA, alpha-linolenic acid (LNA, 18:3n-3), may be an inadequate substitute for DHA because LNA may not be converted to DHA in sufficient amounts to meet an infant's needs. Premature infants lose DHA from their tissues unless they are fed human milk or formula supplemented with DHA. Fish and shellfish are the main food sources of DHA. Women who consume fish have more DHA in their breast milk than do those who do not eat seafood. Infant formulas contain only LNA as a source of n-3 FAs. Pregnant and nursing women should be encouraged to consume seafood on a regular basis during pregnancy and lactation to furnish DHA for their infants. PMID- 8417095 TI - Use of capital budgeting techniques by foodservice directors in for-profit and not-for-profit hospitals. AB - Foodservice directors, who often control one of the largest cost centers in the hospital, are being challenged to manage resources more effectively. Capital budgeting techniques can help enhance a department's cost-effectiveness. The purpose of this study was to assess the use by foodservice directors in nongovernment, not-for-profit and investor-owned, for-profit hospitals of capital budgeting techniques such as payback period, average accounting rate of return, net present value, profitability index, and internal rate of return. Data collected from 84 directors included their use of capital budgeting techniques and operational information about their department. Results indicated that not for-profit hospital foodservices had significantly more full-time equivalent employees than for-profit hospital foodservices--means of 66 and 52, respectively. Size of capital budget was not strongly correlated with any of the operational variables measured. Many of the directors in both types of hospitals used some capital budgeting techniques. However, directors in for-profit hospitals were much more likely than those in not-for-profit hospitals (92% vs 72%) to use capital budgeting techniques. PMID- 8417096 TI - Nutrition attitudes and practices of individuals who are infected with human immunodeficiency virus and who live in south Florida. PMID- 8417097 TI - Nutrition counseling for breast cancer patients. PMID- 8417098 TI - Impact of dietitians on quality of care in 16 nursing homes in east central Indiana. PMID- 8417099 TI - Weight loss program for inner-city black women with non-insulin-dependent diabetes mellitus: PATHWAYS. PMID- 8417100 TI - Striving for empowerment through nutrition education. PMID- 8417101 TI - Position of the American Dietetic Association: nutrition, aging, and the continuum of health care. PMID- 8417102 TI - President's page: 1991-1992 annual report. PMID- 8417103 TI - Dietitians' practices and attitudes regarding the Code of Ethics for the Profession of Dietetics. AB - Three hundred forty-one dietetics practitioners responded (a 34% response rate) to a 103-item questionnaire that included four subscales. This article focuses on one subscale: assessment of the extent to which dietitians reportedly adhere to the profession's code of ethics as it applies to their own practices. The validity of the survey instrument was confirmed by a panel of experts, and the overall internal consistency reliability for the instrument was .82 using Cronbachs' alpha. The internal consistency reliability for the code of ethics subscale was .84 (items 1 through 16) and .88 (items 17 through 20). Respondents to the questionnaire were typically college-educated, married, white women over 40 years of age who practiced as consultant or clinical dietitians. Almost one fourth of the respondents fell into the 60 years and older age group. More than 75% of the respondents reported that they either "always" practiced specific components from the code of ethics or "strongly agreed" with code statements. No statistically significant differences were found among selected personal and professional characteristics and responses to the subscale items. These findings indicate that the dietitians in the sample reportedly adhere to and agree with the Code of Ethics for the Profession of Dietetics. PMID- 8417104 TI - The American Dietetic Association. Officers 1992-1993. Directory of committees. PMID- 8417105 TI - Saponin treatment for in situ hybridization maintains good morphological preservation. AB - The commonly used procedures for in situ hybridization require treatment of the tissue with non-ionic detergents and proteolytic enzymes, resulting in considerable loss of morphological detail. In this study proteinase pre-treatment of the tissue was replaced by saponin, a highly surface-active plant glycoside. This saponin treatment allowed good preservation of tissue morphology, as determined by differential interference and contrast enhanced video microscopy. Saponin pre-treatment resulted in an equal or even better hybridization sensitivity with probes recognizing viral (canine distemper virus) and cellular (myelin) nucleic acid sequences in tissue cultures as well as in paraffin sections. Probable mechanisms of how saponin allows probe penetration while maintaining the morphological details are discussed. PMID- 8417106 TI - Arachidonate 12-lipoxygenase is localized in neurons, glial cells, and endothelial cells of the canine brain. AB - 12-Lipoxygenase oxygenates the 12 position of arachidonic acid, producing its 12S hydroperoxy derivative. We have been interested in the distribution of 12 lipoxygenase in the central nervous system. Previously, by the use of a monoclonal anti-12-lipoxygenase antibody, we proved that one of the arachidonate 12-lipoxygenases existed in canine brain. The present study was therefore designed to elucidate the exact immunohistochemical localization of arachidonate 12-lipoxygenase in canine brain, using a polyclonal anti-12-lipoxygenase antibody. Canine brains were irrigated thoroughly with ice-cold heparinized saline and phosphate-buffered saline to remove blood cells, and then were fixed in 4% periodate-lysine-paraformaldehyde at 4 degrees C for 3 hr. Immunostaining by the indirect method with the peroxidase-labeled antibody was performed for light microscopic observation. The immunohistochemical study demonstrated that many neurons and glial cells in the cerebrum, basal ganglia, and hippocampus were positively stained. Biochemical results as described previously and those of immunohistochemistry indicate that 12-lipoxygenase is definitely localized in various brain parenchymal cells. PMID- 8417107 TI - Application of an image analyzer to gold labeling in immunoelectron microscopy to achieve better demonstration and quantitative analysis. AB - An image analyzer was applied to pre- and post-embedding immunogold labeling with 5-nm gold probes in electron micrographs of several skin basement membrane antigens to improve the visualization of immunolabeling. With a TV camera connected to a color image analyzer, an image of an original immunoelectron micrograph was projected on a TV screen. The image was recorded in the analyzer as Record 1. After the floating threshold method procedure to reduce the contrast of the skin structure, electron-dense 5-nm gold particles could be easily detected. With the analyzer, these particles were then suitably enhanced in color and in size and their image was recorded as Record 2. Records 1 and 2 were then overlapped on the TV screen to build up a double-image picture. Compared with the small, electron-dense 5-nm gold particles in the original electron micrograph, ultrastructural localization of bullous pemphigoid antigen, epidermolysis bullosa acquisita antigen, and the collagenous part of Type VII collagen were more clearly and easily seen, even on low-magnification electron micrographs. The level of background labeling could also be accurately and objectively evaluated. By counting the number of gold particles labeling a certain area and using the analyzer to interpret the result as a diagram, quantitative analysis was also possible. We believe that this technique should be widely applicable to immunogold electron microscopy, not only of skin but also of other substrates of interest. PMID- 8417108 TI - Divergent and co-localization of the two small proteoglycans decorin and proteoglycan-100 in human skeletal tissues and tumors. AB - The core protein of a recently described small proteoglycan, proteoglycan-100, was localized in fetal human tissues by indirect immunocytochemistry and compared with the localization of known members of the small proteoglycan family. Co localization of decorin and proteoglycan-100 was seen in bone tissue but decorin and proteoglycan-100 exhibited a substantially divergent distribution in fetal skin, cartilage and in the mineralization zone of the growth plate. Proteoglycan 100 was also found in striated muscle, nerve fibers, and synovial tissue. Immunostaining of a chondroblastic osteosarcoma demonstrated chondroid cells selectively expressing either proteoglycan-100 or decorin. Co-expression of both small proteoglycans was observed in sections from a chordoma. In fetal bone and in the two tumors, colocalization of proteoglycan-100 and of biglycan was also found. These results provide evidence of the wide and characteristic distribution of proteoglycan-100. PMID- 8417109 TI - Immunolocalization of inhibin in the mammary gland of rats treated with hCG. AB - Human chorionic gonadotropin (hCG) induces profuse lobular development in the mammary gland of young virgin rats. To clarify whether the effect of hCG is locally mediated by inhibin, a non-steroidal glycoprotein, we detected its localization immunocytochemically in the mammary gland with polyclonal antibodies against the alpha- and beta-chains. Virgin female Sprague-Dawley rats were sacrificed after treatment with a daily IP injection of 100 IU hCG for 5, 10, 15, or 21 days of treatment or 20 days after the last injection. Whereas the mammary gland of control animals did not contain immunoreactive inhibin, hCG treatment induced the expression of inhibin in the cytoplasm of alveolar cells but not in ductal cells. The reaction became evident by Day 10 of treatment and reached its maximal intensity by Day 15. Thereafter, the reaction became evident in the stroma, which exhibited maximal positivity by Day 20. Once hCG treatment was terminated, the mammary gland regressed to its pre-treatment condition, appearing similar both in morphology and inhibin content to that of control animals. The expression of this glycoprotein hormone in the mammary gland after hCG administration at the time of maximal lobulolveolar development, and its diffusion towards the stroma during regression, suggest a critical role of inhibin as a modulator of mammary growth and differentiation. PMID- 8417110 TI - Novel monoclonal antibodies specific for human cardiac myosin light-chain 1: useful tools for analysis of normal and pathological hearts. AB - To investigate the developmental, physiological and pathophysiological roles of human cardiac myosin light-chain 1 (LC1s), we developed two novel monoclonal antibodies (KA1 and KB1) against human cardiac LC1s and examined LC1s in normal and pathological hearts immunohistochemically. KA1 and KB1 were specific only for atrial LC1 (ALC1) and for both ALC1 and ventricular LC1 (VLC1), respectively, in human hearts. Among human tissues tested, including skeletal muscle, vascular smooth muscle, and liver, KA1 did not crossreact with proteins in any other tissues than atria, whereas KB1 crossreacted with the slow-type LC1 of skeletal muscle. Among adult mammalian hearts of several other species including pig, dog, hamster, and rat, KA1 and KB1 crossreacted only with ALC1 and with both ALC1 and VLC1, respectively. ALC1 was strongly and uniformly observed in human fetal atria and ventricles and in normal adult human atria, but sporadically in normal adult human ventricles. In the overloaded ventricle (dilated cardiomyopathy), ALC1 was highly augmented but not uniform. These results suggest that the fetal VLC1 is immunohistochemically identical to the adult type of ALC1 and that ALC1 is expressed homogeneously in human fetal ventricles and sporadically in normal adult ventricles, and is re-expressed heterogeneously and in an increased amount in the overloaded ventricle. PMID- 8417111 TI - Densities of NADPH-ferrihemoprotein reductase and cytochrome P-450 molecules in the endoplasmic reticulum membrane of rat hepatocytes. AB - In hepatocytes, NADPH-ferrihemoprotein reductase (reductase) has been hypothesized to exist as aggregates or micelles in endoplasmic reticulum (ER) membrane. However, if the number of reductase molecules per unit area of ER is low, this hypothesis cannot explain how a few reductase molecules efficiently reduce many P-450 molecules. To test this hypothesis, we estimated the numbers of reductase and P-450 molecules per unit ER area (reductase and P-450 densities) by microphotometry of the two enzymes in conjunction with morphometry of ER in periportal, midzonal, and perivenular rat hepatocytes. The reductase density in periportal, midzonal, and perivenular hepatocytes (107-179 molecules/microns 2 of ER) was high enough to efficiently reduce all P-450 molecules in the ER, although the value in perivenular hepatocytes was lowest owing to the relatively greater amount of ER in this region. The pattern of sublobular gradient in the reductase density was similar to that in the P-450 density. Consequently, the molar ratio of P-450 to reductase in ER was similar (about 40:1) in hepatocytes regardless of their positions within the liver lobule. PMID- 8417112 TI - Ultrasound-amplified immunohistochemistry. AB - We describe a novel technique for improving the sensitivity of immunofluorescence staining by use of ultrasonic irradiation. Free-floating vibratome sections from rat cerebellum were incubated with primary antiserum and simultaneously were briefly exposed to ultrasound (US) in a conventional ultrasound bath. After the US treatment, a conventional immunohistochemical method was employed. Two different antisera and two conventional immunohistochemical detection systems were tested. In all cases a 10-20-sec US treatment strengthened immunoreactivity considerably. Irradiated samples showed a good morphology compared with non irradiated sections. Our results demonstrate that with ultrasonic treatment the dilutions of primary antibodies can be increased and the incubation times of primary antisera can be reduced. The ultrasonic method described here requires no special equipment. It is easy, reproducible, and it can be considered a new method for the enhancement at immunohisto- and cytochemical staining of free floating vibratome sections. PMID- 8417113 TI - Internalization of surfactant protein A (SP-A) into lamellar bodies of rat alveolar type II cells in vitro. AB - Pulmonary surfactant is thought to be internalized and processed for reuse by alveolar Type II cells. In the present study we followed the internalization and intracellular trafficking of purified surfactant protein A (SP-A) by primary cultures of alveolar Type II cells. Internalization of native rat SP-A was compared with that of recombinant rat and human SP-A isolated from a patient with alveolar proteinosis. All SP-A species were conjugated with colloidal gold for visualization by electron microscopy. The gold conjugates were biologically active, as demonstrated by inhibition of phospholipid secretion from alveolar Type II cells. The SP-A-gold conjugates were internalized to lamellar bodies (LB) via the endosomal system, which included both electron-lucent and -dense multivesicular bodies. Labeling of LB was time dependent, and after 7 hr 30-40% of these organelles were labeled. Alkylation of SP-A greatly reduced internalization, as did an excess of non-conjugated SP-A. No qualitative differences in uptake were observed with the three forms of SP-A. The percent of labeled LB was similar (30-40%) after 7 hr of internalization with the three species of SP-A. The recombinant SP-A produced using a baculovirus vector lacked hydroxyproline and had an altered oligosaccharide, but these features did not affect its internalization or the rate of LB labeling. Internalization of the gold-conjugated SP-A and endocytosis of the fluid-phase marker Lucifer Yellow were related to the shape of Type II cells. Both uptake of SP-A, which is receptor mediated, and fluid-phase endocytosis were found to be less active in the flattened than in the rounded cells. Therefore, cell shape and hence cytoskeletal organization may play an important role in SP-A recycling. However, it is possible that both morphology and decreased endocytosis are independent manifestations related to the loss of differentiated function of cultured Type II cells. PMID- 8417114 TI - Secretion of glandular kallikrein and renin from the basolateral pole of mouse submandibular duct cells: an immunocytochemical study. AB - Glandular kallikrein from salivary glands in rats has been measured in the circulation and has been shown to have local vasoactive effects. In mice, renin and epidermal growth factor from the submandibular gland (SM) also reach the circulation, as both have been measured in plasma. The route by which these peptides enter the blood from their site of synthesis in ducts of the SM is unclear. We have investigated by immunocytochemistry the secretory pathways for kallikrein and renin from salivary duct cells in mice. The renal/pancreatic kallikrein-secreting cells of the striated and excretory ducts of the SM were distinguished from the granular convoluted tubule (GCT) cells which secrete other glandular kallikreins on the basis of data obtained in previous studies, in which we used gene-specific oligonucleotide probes to identify the expressing cell types. Renal/pancreatic kallikrein was apparently secreted constitutively from the basolateral surface of striated duct cells and in secretory vesicles from excretory duct cells, whereas apical secretion occurred via the regulated pathway in both cell types. Glandular kallikreins and renin synthesized in GCT cells were secreted from the basolateral surface by dissolution of granules at the cell membrane. There were fenestrated capillaries underlying the duct tree which would enable the secreted products to reach the circulation. PMID- 8417115 TI - Functional consequences of engagement of the T cell receptor by low affinity ligands. AB - The mechanisms involved in TCR antagonism by Ag analog/MHC have been analyzed. A detailed structure-activity relationship study indicated that modification of any of the major T cell contact residues of the peptide molecule can yield a powerful antagonist. It was also shown that as the analog structure increased in similarity to the Ag, the capacity to antagonize Ag-TCR interaction increased up to the point that the analogs themselves became antigenic. These data strongly suggest an affinity-related mechanism whereby a certain affinity is required for signaling through the TCR, and that below this level there can be sufficient affinity to engage the receptor such that triggering does not occur and antagonism can be detected. Taking advantage of this information, antagonist peptides active down to the 10 nM range were engineered. Thus, this approach demonstrates for the first time a rational approach to designing effective, selective low m.w. compounds with high potential in treatment of allergies and autoimmune diseases. PMID- 8417116 TI - Cytokine synthesis by intestinal intraepithelial lymphocytes. Both gamma/delta T cell receptor-positive and alpha/beta T cell receptor-positive T cells in the G1 phase of cell cycle produce IFN-gamma and IL-5. AB - Murine intestinal intraepithelial lymphocytes (IEL) have been shown to contain subsets of alpha/beta TCR+ and gamma/delta TCR+ T cells that spontaneously produce cytokines such as IFN-gamma and IL-5. We have now determined the nature and cell cycle stage of these cytokine-producing T lymphocytes in EIL by using IFN-gamma- and IL-5-specific ELISPOT assay, cytokine-specific mRNA-cDNA dot-blot hybridization and polymerase chain reaction, and flow cytometry (FACS) for DNA analysis. When CD3+ T cells from IEL of normal C3H/HeN mice were separated into low and high density fractions by discontinuous Percoll gradients, IFN-gamma and IL-5 spot-forming cells were only found in the former population. Analysis of mRNA for these cytokines by both IFN-gamma- and IL-5-specific dot-blot hybridization and polymerase chain reaction revealed that higher levels of message for IFN-gamma and IL-5 were also seen in the low density fraction. However, cell cycle analysis of these two fractions by FACS using propidium iodide showed a similar pattern of cell cycle stages in both low and high density populations (G0 + G1 approximately 96 to 98% and S/G2 + M approximately 2 to 4%). Finally, mRNA from gamma/delta TCR+ and alpha/beta TCR+ T cells in both low and high density fractions of IEL were analyzed for IFN-gamma and IL-5 message by polymerase chain reaction. After 35 cycles of amplification, both gamma/delta TCR+ and alpha/beta TCR+ T cells in the low density fraction expressed higher levels of message for these two cytokines when compared with the high density population. These results have now shown that both gamma/delta and alpha/beta TCR+ IEL can be separated into low and high density subsets and both fractions possess a similar stage of cell cycle. However, only the low density cells (in G1 phase) of both gamma/delta and alpha/beta TCR types possess increased cytokine specific mRNA and produce the cytokines IFN-gamma and IL-5. Our results suggest that alpha/beta TCR+ and gamma/delta TCR+ IEL can produce cytokines without cell proliferation. PMID- 8417117 TI - Alternatively spliced pp52 mRNA in nonlymphoid stromal cells. AB - The 52-kDa phosphoprotein, also reported as lymphocyte-specific gene 1 and WP34, is transcribed as a 1.6-kb mRNA in B lymphocytes, B cell lines, and untransformed T cells. This gene encodes a cytoplasmic and plasma membrane-associated protein that is phosphorylated at a casein kinase II site and reportedly binds calcium. Based on these properties, it has been hypothesized that lymphoid form of the 52 kDa phosphoprotein protein may play a role in lymphocyte signal transduction. We show that alternatively spliced mRNA are expressed from this gene in nonlymphoid cell lines (myocytes, stromal cells, fibroblasts). These cell lines do not express the 1.6-kb lymphoid cell-specific transcript. Instead, mRNA of 2.0 and 2.8 kb are detected in varying abundance. A full-length 2.0-kb cDNA has been cloned and sequenced from the BMS2 stromal cell line by conventional screening and polymerase chain reaction-based methods. This cDNA clone, designated S37, has a single open reading frame encoding a 328 amino acid peptide. The nucleotide sequence of the S37 stromal cell cDNA is identical to that of the lymphocyte derived pp52 cDNA from the 3' poly(A) tail to the codon encoding the amino acid at residue 24. This region of the S37 cDNA clone encodes a protein that is identical to that encoded by the lymphoid pp52 cDNA and includes a casein kinase II phosphorylation site. However, the two clones differ in their 5' nucleotide sequence and their NH3 terminal amino acid sequence. This organization is consistent with alternative exon utilization. These results suggest that tissue specific control mechanisms are used to generate different forms of lymphoid form of the 52-kDa phosphoprotein mRNA in lymphoid cells versus mesoderm-derived, nonlymphoid cell lineages. PMID- 8417118 TI - T cell antigen receptor-eta subunit. Low levels of expression and limited cross species conservation. AB - The eta-subunit of the TCR is derived from an alternative splice product of the TCR-zeta gene. The eta-subunit has been extensively characterized in murine T cells, where up to 10% of TCR bear zeta-eta heterodimers rather than zeta homodimers. In contrast to the significant levels of eta found in murine T cells, in human PBL an eta-like region is expressed spliced to upstream exons of the zeta-gene at no more than 0.25% the level of zeta-mRNA. Analysis of genomic DNA from five additional mammalian species demonstrated that eta-like sequences are highly conserved at a nucleic acid level. The protein sequences encoded in the eta-regions from various species ranged from 28 to 92 amino acids in length, with the first seven deduced amino acids of eta-being common to all species. Within three to five amino acids of this region, all species have five consecutive charged amino acids. Beyond this point, due to translation in different reading frames, there is no significant amino acid homology. The findings of low level of expression of eta-RNA, limited cross-species conservation on a protein level, and the lack of an established functional role for this alternative splice product raise questions as to the potential roles that this subunit may play in TCR function. PMID- 8417119 TI - Effector functions of a mouse IgG that lacks the entire CH1 domain. C1q binding and complement fixation in the absence of antigen. AB - C1q binding and complement fixation were examined of a short-chain IgG2a antibody that lacks the entire CH1 domain. This short-chain antibody has been reported to have a low level of constitutive complement-activating activity in the absence of Ag. Two-dimensional SDS/PAGE and cation-exchange chromatography have demonstrated that two types of IgG2a proteins are secreted by the short-chain IgG2a antibody producing cell line. It has been shown that 1) the difference between these two types of the IgG2a proteins is whether the two L chains are linked by a disulfide bridge or not, and 2) C1q-binding and complement-activating activities are expressed only when the inter L chain disulfide bridge does not exist. A molecular model is presented for the active form of the short-chain IgG2a antibody. PMID- 8417120 TI - Multiple cis-acting elements are required for proper transcription of the mouse V delta 1 T cell receptor promoter. AB - To gain insight into the developmentally regulated expression of the mouse TCR V delta-gene segments, we have investigated the role of the 5' promoter region of the V delta 1-gene. Transient transfection assays showed that a construct encompassing 267 nucleotides upstream from the mapped transcriptional start site was capable of driving promoter activity when transfected into V delta 1+ T cells. The inclusion of an additional 459-bp 5' segment to this construct did not affect promoter activity. However, a deletion of 222 5' nucleotides from the same construct dramatically decreased promoter activity. In vivo genomic footprinting localized several protein-DNA interactions to the stretch of DNA shown to have transcriptional activity. A computer analysis revealed that the segments of DNA participating in these protein-DNA interactions were identical to the previously described cyclic AMP response element (CRE), E box, and leukemia virus E26 cis acting elements. Transient transfection assays performed with -267 bp constructs containing mutations at each of the localized cis-acting elements revealed that the CRE, E box, and Ets elements work together in driving promoter activity and that the CRE and Ets elements are the most important for driving transcription. Gel mobility shift analyses showed that each of these cis-acting elements is capable of binding specific nuclear factors present in V delta 1-expressing cells. These data indicate that multiple transcription factors acting in concert are responsible for V delta 1 gene expression. PMID- 8417121 TI - Expression pattern of the most JH-proximal human VH gene segment (VH6) in the B cell and antibody repertoire suggests a role of VH6-encoded IgM antibodies in early ontogeny. AB - We have developed a mAb (JE-6) that recognizes an Id encoded by the most JH proximal human VH gene segment (VH6) in or near germ-line configuration. This mAb was used to determine the frequency of Id JE6+ B cells in large collections of monoclonal EBV-transformed and short term B cell lines derived from fetal, neonatal, and adult lymphoid tissues. Moreover, we investigated the presence of Id JE-6+ Ig in sera from neonates and adults and determined the (auto)antigen binding properties of VH6-encoded IgM mAb. We detected a fivefold overrepresentation of VH6-expressing IgM producing B cells in fetal tissues, cord blood, and adult bone marrow relative to adult blood. In cord blood, but not in adult blood sera, germ-line VH6-encoded IgM molecules were readily detectable. IgM secreted by VH6-expressing B cell clones displayed highly conserved and virtually identical autoantigen binding properties, independent of the length and composition of the IgH chain CDR3 region and L chain isotype. Collectively, these results suggest that the VH6 gene and the antibodies it encodes play an important role in early human ontogeny. PMID- 8417122 TI - Human serum amyloid P component oligomers bind and activate the classical complement pathway via residues 14-26 and 76-92 of the A chain collagen-like region of C1q. AB - Serum amyloid P component (SAP) was polymerized using the cleavable cross-linker 3,3'-dithio-bis-(sulfo-succinimidylpropionate) to study its interaction with the C system. Dimers and trimers, but no larger oligomers, were observed; the trimers retained native SAP immunoreactivity (except for one calcium-dependent epitope) without displaying neo-SAP epitopes. The SAP trimers bound strongly to C1q, at the level of the collagen-like region (CLR). SAP bound to synthetic C1q A chain peptides 14-26 and 76-92, and these peptides inhibited the binding of SAP trimers to the CLR. When incubated in dilute human serum, SAP trimers consumed total C and C4, but not alternative pathway, hemolytic activities. Consumption of C4 by SAP trimers was inhibited by C1q A chain peptide 14-26. Thus, SAP oligomers bind C1q and activate the classical C pathway via the collagen-like region of C1q, at sites located within residues 14-26 and/or 76-92 of the C1q A chain. PMID- 8417123 TI - Intratumor gene expression after adoptive immunotherapy in a murine tumor model. Regulation of messenger RNA levels associated with the differential expansion of tumor-infiltrating lymphocytes. AB - The aim of this paper was to show that the adoptive immunotherapy (AIT) of established tumors resulted in the activation of defined lymphocyte-associated genes at the site of rejection. C57BL/6J mice bearing the moderately immunogenic syngeneic MCA/76-9 sarcoma received combination therapy 10 days after tumor cell implantation. This consisted of a single i.p. injection of cyclophosphamide (CY) followed by the i.v. injection of tumor-sensitized T cells (CY/AIT). The previously observed in situ differential expansion of tumor-infiltrating lymphocytes (TIL) was associated with a parallel modulation of CD4 (L3T4) and CD8 Ag expression. Flow cytometric analysis indicated that the CD8+ TIL appeared to contain two subpopulations during the reported proliferative phase, but by day 8 after CY/AIT the cells were composed of a single bright population. The CD4+ TIL similarly appeared to show two subpopulations, but in contrast to the CD8+ TIL there was a shift to a predominantly less bright single population by day 8. The expression of lymphocyte genes (Ly-2 and Ly-4 encoding the CD8 and CD4 Ag, respectively, IL-2, IL-2R, IFN-gamma, and IL-6) was analyzed by Northern hybridization using RNA extracted from whole tumor tissue. Progressing tumors expressed only low and relatively constant levels of mRNA for all of the genes except Ly-2 over a 19-day period. In contrast, there were considerable temporal fluctuations in mRNA levels depending on whether the mice had received CY or CY/AIT, but it was apparent that CY/AIT induced the more dramatic changes as far as Ly-2, Ly-4, IL-2, and IFN-gamma mRNA levels were concerned. A preliminary survey of cytokine bioactivity released by tumor-associated cells isolated 9 days after CY/AIT indicated that both IFN-gamma and IL-6 activities were released by the cells, but not IL-2, despite the relatively high levels of IL-2 mRNA. These data provide evidence that the differential expansion of TIL after CY/AIT is accompanied by well defined changes in the levels of mRNA-encoding TIL membrane Ag and lymphokine genes and are concordant with the view that amplified anti tumor immune responses occur after AIT. PMID- 8417124 TI - Generation of diacylglycerol and ceramide during homologous complement activation. AB - Formation of sublytic terminal complement complexes (TCC) on nucleated cells produces transient increase in [Ca2+]i and activates protein kinase C. The present study is to evaluate whether TCC can generate endogenous signal messengers other than Ca2+ that regulate cell activities by measuring mass-levels of sn-1,2-diacylglycerol (DAG) and ceramide. As targets, lymphoblastoid human B cell lines JY25 and its mutant JY5 were used. JY5, cells deficient in glycosylphosphatidylinositol-anchored proteins with higher lytic susceptibility to human complement, are four times more efficient in forming C5b-9. When cells sensitized with limited anti-class II IgG were exposed to human serum to generate sublytic TCC, a sustained increase in DAG and ceramide was observed with a maximum 3.6-fold DAG increase over basal level in JY25 and 2.8-fold in JY5, and 6.3-fold ceramide increase in JY25 and 2.8-fold in JY5. The effect of TCC was evaluated with C7-deficient human serum (C7D) +/- C7 and also with C5b6, C7, C8, and C9 proteins. The DAG and ceramide increase by C7D + C7 over C7D control were 1.6- and 1.8-fold, respectively, in JY25, and 2.3-, and two-fold in JY5. TCC activation also induced an increased hydrolysis of sphyingomyelin and phosphatidylcholine. In addition, DAG increase by TCC was primarily achieved by C5b-7 and preincubation of cells with pertussis toxininhibited DAG increase, suggesting an involvement of a pertussis toxin-sensitive GTP-binding protein. As important signal transduction molecules, DAG and ceramide generated in response to TCC assembly, could participate in cell activation during inflammation and repair. PMID- 8417125 TI - Depression of macrophage respiratory burst capacity and arachidonic acid release after Fc receptor-mediated phagocytosis. AB - The phagocytosis of IgG-coated erythrocytes (ElgG) by macrophages results in a subsequent depression of macrophage phagocytic function, respiratory burst capacity, and bactericidal activity. Our study was carried out to determine the importance of impaired arachidonic acid release in the depression of the respiratory burst after ElgG phagocytosis. The depression of triggered H2O2 production after ElgG phagocytosis was not due to cyclooxygenase products because indomethacin or aspirin did not modify the depression. Further studies revealed that the depression of triggered H2O2 production after ElgG phagocytosis was associated with a depression in the ability of macrophages to release arachidonic acid in response to PMA, zymosan, or calcium ionophore. The addition of exogenous arachidonic acid partially prevented the depression of triggered H2O2 production after ElgG phagocytosis. Unlike phagocytosis mediated by FcR, complement receptor mediated phagocytosis did not alter H2O2 production or arachidonic acid release. Ligation of FcR was not sufficient to depress triggered H2O2 production and arachidonic acid release because these functions were not depressed when phagocytosis was inhibited with cytochalasin D. Thus, it was found that the depression of triggered H2O2 production by macrophages after FcR-mediated phagocytosis was associated with impaired release of arachidonic acid and that H2O2 production could be partially restored by the addition of arachidonic acid. These results suggest that the impairment of arachidonic acid release after FcR mediated phagocytosis contributes to the depression of macrophage respiratory burst capacity after FcR-mediated phagocytosis. PMID- 8417126 TI - Relationship of C5a receptor modulation to the functional responsiveness of human polymorphonuclear leukocytes to C5a. AB - The relationship of C5a receptor expression on human polymorphonuclear leukocytes (PMN) to the functional response of these cells to C5a was studied using flow cytometry. C5a receptor expression was determined with a fluorescein conjugate of C5a and oxidative burst activity was monitored by conversion of dichlorofluorescein to dichlorofluorescein (DCF) as a measure of H2O2 production. These studies showed that after incubation of PMN with increasing concentrations of C5a, and allowing for internalization of bound ligand, more than 40% of the cell surface C5a receptors were internalized before the DCF response to optimal concentrations of C5a was decreased below the levels for untreated control cells. Although C5a responsiveness was lost after preincubation with 10(-8) M C5a, cells remained responsive to formyl peptide. In other studies, cells were preincubated with unlabeled C5a under conditions that provided for internalization of nearly all C5a receptors. PMN were then cultured for up to 90 min and monitored for C5a receptor reexpression and return of cell function. In these studies, the DCF response of PMN to C5a returned to 100% much earlier than the cells regained full expression of C5a receptors. The DCF response to formyl peptide remained intact throughout the period of C5a receptor reexpression. These studies showed that once > 40% of the original population of C5a receptors are reexpressed on the PMN, that these cells regain 100% of their functional responsiveness to C5a in the DCF assay. Evaluation of the affinity and number of C5a receptors using 125I labeled C5a after receptor reexpression showed that maximal receptor reexpression was approximately 73% of that obtained with control cells and the Kd of reexpressed receptors was 0.60 vs 0.94 nM for control cells. These studies demonstrate that only a portion of the total C5a receptors expressed on PMN are essential to stimulate a 100% functional response in PMN and that the reexpressed receptors are capable of transducing a signal that activates the oxidative burst in these cells. PMID- 8417127 TI - Nonmitogenic anti-CD3F(ab')2 fragments inhibit lethal murine graft-versus-host disease induced across the major histocompatibility barrier. AB - We have investigated the in vivo administration of nonmitogenic anti-CD3F(ab')2 fragments for the prevention of lethal graft-vs-host disease (GVHD) in irradiated recipients of fully allogeneic bone marrow cells plus splenocyte (BMS) inocula. Recipients of anti-CD3F(ab')2 fragments administered for 1 mo post-bone marrow transplantation (BMT) had 100% survival without clinical or histopathological evidence of GVHD. Controls given saline injections succumbed by 39 days post-BMT. Similar results were obtained in groups of recipient mice given BMS in which T cells were depleted by in vitro anti-Thy-1.2 plus C' treatment. Further studies were undertaken to define mechanistic differences in the two approaches. Using Ly 5 congenic sources of donor bone marrow and spleen, we determined that anti CD3F(ab')2 fragments induced TCR modulation and T cell depletion. Mature splenic derived CD4+ cells were depleted to a greater extent than CD8+ cells. Early post BMT, recipients receiving injections with control saline had the highest number of CD4+ and CD8+ cells (which may cause GVHD) followed by recipients of anti CD3F(ab')2 fragments, with the fewest CD8+ cells observed in the anti-Thy-1.2 + C' treated group. CD3+CD4-CD8- cells (which may suppress GVHD generation) were present in higher numbers early post-BMT in recipients given anti-CD3F(ab')2 fragments as compared to recipients given anti-Thy-1.2 + C'-treated BMS. In long term survivors, a mononuclear T cell containing infiltrate without evidence of destruction was observed in sites of GVHD (lung and liver), consistent with a "Quilty" effect, which was not observed in either of the other two groups. Although survivors were tolerant of donor skin grafts and rejected third party grafts, recipients given anti-CD3F(ab')2 fragments but not anti-Thy-1.2 + C' treated BMS had vigorous anti-host proliferative responses. These results demonstrate that although in vitro anti-Thy-1.2 + C' treatment of BMS (which is highly depletionary) and in vivo administration of anti-CD3F(ab')2 fragments (which is modulatory and less depletionary) are both effective strategies for GVHD, the cellular events involved in achieving GVHD prevention are indeed different. PMID- 8417128 TI - Selective transformation of host lymphocytes in vivo by retrovirus-producing macrophages. AB - Macrophage-mediated retroviral transformation of host cells was studied in vivo utilizing the cloned murine macrophage-line GG2EE, generated by in vitro infection of bone marrow cells from C3H/HeJ mice (H-2k) with the acute transforming retrovirus J2 bearing the v-myc and v-raf oncogenes. Because GG2EE macrophages produce the J2 retrovirus, the development of secondary, J2 virus induced tumors after the injection of the cell line into several strains of mice was evaluated. GG2EE cells proliferated and gave rise to histiocytic tumors in syngeneic mice and in allogeneic athymic Swiss mice. The inoculum of GG2EE cells in allogeneic DBA/2 mice (H-2d) and, to a lesser extent, in BALB/c (H-2d) and BALB/k (H-2k) mice gave rise to a small, solid mass at the injection site. Although the initial tumor was slowly rejected, secondary lymphomas belonging to the B or T cell lineage developed, leading to mouse death. Extensive phenotypic, functional, and chromosomal analyses proved that lymphomas were derived from host T and B cell transformation. Southern and Northern blot studies showed that J2 virus was integrated and expressed in lymphoma cells, demonstrating that the virus was transmitted to the host lymphocytes and suggesting that it was causal in lymphoma development. The existence of close and protracted interactions between GG2EE macrophages and allogeneic host lymphocytes and the presence of viral particles in the area of macrophage-lymphocyte contact were demonstrated by histologic and ultrastructural analysis. Rejection of J2 virus-infected lymphocytes in allogeneic mice suggested that host lymphocyte transformation was dependent upon the macrophage cell type. These results demonstrate that macrophage-derived J2 retrovirus transforms host lymphocytes in vivo in allogeneic mice and that a condition of host alloreactivity is critical for such event. PMID- 8417129 TI - Follicular dendritic cells and B cell costimulation. AB - Ag-bearing follicular dendritic cells (FDC) are found throughout secondary lymphoid tissues in close association with rapidly proliferating germinal center B lymphocytes. We reasoned that FDC might provide costimulatory signals that would enhance the ability of Ag to stimulate B cell proliferation in the germinal centers. To test this, FDC were cultured with B cells activated by a slg dependent (goat anti-mouse mu conjugated to dextran (anti-mu-dex)) or independent (LPS) pathway and their proliferation was measured by using [3H]thymidine incorporation. The addition of FDC markedly augmented B cell proliferation in a dose-dependent fashion. Depletion of FDC from cultures abrogated the increased proliferation. Addition of highly purified FDC obtained from cell sorting resulted in B cell costimulation, whereas addition of other sorted cells was without effect. The FDC accessory activity was apparent over the entire culture period and over a wide range of either polyclonal B cell activator. When B cells and activators were cultured in the absence of FDC, only about one fourth of the cells remained viable after 3 days. In contrast, virtually all cells in cultures containing FDC, B cells, and activator were viable. Cultures containing FDC and B cells from nude mice proliferated normally in the presence of anti-mu-dex plus rIL-4, implying that IL-4 provides adequate T cell help in this system. The costimulatory activity of the FDC could not replace either the anti-mu-dex or IL-4 in this system and was not MHC restricted. These data support the concept that FDC not only provide Ag but also facilitate B cell proliferation by means of other costimulatory interactions that contribute to make the microenvironment in the germinal center favorable for B cells to proliferate. PMID- 8417130 TI - Killer cell recruitment and renewal capacity of purified cytolytic and noncytolytic human peripheral blood natural killer cell subsets. AB - The inability to isolate NK precursors at different stages of development has impeded understanding of the processes involved in NK maturation. The present studies utilize a flow cytometric technique that enables the isolation of operationally defined cell subsets, lytic (killers) and nonlytic conjugate forming (binders), and nonconjugate-forming (free) cells, within NK-enriched preparations to test whether these might represent cells in different stages of NK development. To characterize both the steps involved in NK maturation and the cells responsible for IL-2 induced proliferation, these purified subsets were analyzed for their killer cell recruitment and renewal capacity. After a 2-h exposure to IFN-alpha or IL-2, induction of lytic function was developed only in the binder subset as detected in the single cell assay. Neither enhancement of killer cell recycling nor induction of binding function among the subsets was observed. However, after an 18-h culture period, with or without rIL-2, killer cells preferentially expressed activation Ag CD69 (Leu-23) and the IL-2R alpha chain, TAC (CD25). In addition, all cells in contact with K562 targets displayed enhanced expression of these Ag. Killer cells also showed an enhanced capacity to proliferate in response to rIL-2 in a 6-day [3H]TdR incorporation assay. Additional irradiated K562 targets enhanced the proliferative capacity of all the subsets, with only a marginal effect on sorted free cells. Nevertheless, sorted free cells, in addition to binders, developed potent binding and lytic function when tested in the single-cell assay after 4 days of IL-2 culture. The lytic activity of killers was reduced, as compared with freshly isolated killers. The results are consistent with a two-step model for NK maturation, involving the acquisition of lytic function before proliferative capacity, and specific triggering of killer cells through interaction with target cells for induction of a proliferative competent state. PMID- 8417131 TI - Fresh and cultured Langerhans cells display differential capacities to activate hapten-specific T cells. AB - Langerhans cells (LC) that have been cultured for 3 days acquire potent T cell activating properties when compared to freshly prepared, uncultured LC. By contrast, fresh LC are superior to cultured LC in the ability to process native protein Ag. To define further the disparate functional properties of these epidermally derived APC, freshly isolated and cultured epidermal cells (EC) enriched for LC were prepared from BALB/c mice. Highly purified T cells from naive mice, and from mice sensitized epicutaneously with dinitrofluorobenzene, have been examined for their capacity to respond to fresh and cultured EC; 1) in the presence of staphylococcal enterotoxin B; and 2) after the EC had been derivatized with dinitrofluorobenzene. Both fresh and cultured EC activated syngeneic T cells in the presence of staphylococcal enterotoxin B, and fresh and cultured DNP-derivatized EC induced proliferation among DNP-specific T cells. Only cultured, hapten-derivatized EC were able to activate unprimed syngeneic T cells in vitro, and these cells responded as though "primed" when re-exposed to DNP-derivatized spleen cells in secondary cultures. In addition, naive lymphocytes that were activated by cultured DNP-EC were able to evoke local contact hypersensitivity reactions when injected into the pinnae of naive mice that were then painted with dinitrofluorobenzene. By contrast, naive syngeneic T cells exposed to fresh DNP-EC neither proliferated nor differentiated into effector cells. We conclude that fresh LC can constitutively activate primed, but not unprimed, hapten-specific T cells, whereas cultured LC readily both primed and unprimed T cells. The capacity of hapten-derivatized cultured EC to convert naive, hapten-specific T cells into cells that mediate contact hypersensitivity supports the proposal that cultured LC are the functional equivalents of epidermal LC that have migrated to draining lymph nodes. The ability of hapten derivatized fresh LC to activate primed, hapten-specific T cells is consistent with the view that fresh LC are functionally equivalent to LC within the epidermis. PMID- 8417132 TI - Production of DNP-specific/class I MHC-restricted suppressor molecules is linked to the expression of T cell receptor alpha- and beta-chain genes. AB - The hapten/class I MHC-specific soluble immunoregulatory molecules produced by CD8+T cells from dinitrobenzene sulfonate-primed mice express the binding specificity and serologic determinants of alpha/beta TCR. To examine the genes used to encode these soluble immunoregulatory molecules, we utilized a surface TCR expressing Ts hybridoma, which constitutively produces a DNP/Kd-specific regulatory molecule. Northern and Southern analyses indicated that MTs 79.1 cells use a V beta 8 and a V alpha 4 gene to encode the variable regions of the surface alpha/beta TCR. A panel of TCR- variants was generated by subjecting MTs 79.1 cells to gamma-irradiation. Twelve of the TCR- variants were chosen for detailed characterization. Northern blot analyses indicated the absence of the MTs 79.1 V alpha 4 chain transcript in five of the variants and the absence of the parental V beta 8 chain transcript in the other seven. Southern blot analyses demonstrated the deletion of the parental gene encoding the alpha- or beta-chain from the genome of the respective mutant. None of the 12 TCR gene deletion mutants produced the parental suppressive activity. Expression of the parental TCR beta chain gene in one of the beta-chain gene deletion mutants reconstituted the ability to produce this activity. As with the MTs 79.1 molecule, the regulatory molecule produced by the beta-chain gene transfectant was bound by and eluted from Sepharose columns coupled with either DNP or anti-V beta 8 antibodies. These results establish a strong linkage between the suppressor molecules produced by these Ts and TCR alpha- and beta-chain gene transcription. PMID- 8417133 TI - Delimitation of the proliferative stages in the human thymus indicates that cell expansion occurs before the expression of CD3 (T cell receptor). AB - Precursor of T lymphocytes undergo proliferation and maturation under the influence of the thymic microenvironment. In our study, we have attempted to determine the distribution of human postnatal thymocytes in division according to their stage of differentiation. Our data show that about 11.5% of all thymic cells are in S/G2/M phases, and that a subset of the cortical and precortical subpopulations contains most of the dividing cells. Rate of cell division is maintained at high levels from the prethymocyte precursor along the successive stages of differentiation represented by CD1+CD3-CD4-CD8- and CD1+CD3-CD4+CD8- cells. The percentage of dividing cells is maximal in an intermediate subset of CD1+CD3-CD4+CD8-CD45RO+ cells defined by the distinct expression of class I HLAdim/high molecules, which could contain cells in transit from prethymocytes to double-positive cortical cells. The CD3- fraction of the double-positive cortical cells contains most of the dividing thymocytes, although the rate of division within this subset is much less than that of the precursor CD1+CD3-CD4+CD8- cells. In a linear scheme of differentiation, cell division stops at or near the point of initiation of CD3 expression. These results suggest that in human thymus cell expansion takes place before the initiation of the positive selection process. According to this view the stringency of the selection process would require the previous generation of a large number of precursors to permit the production of sufficient numbers of mature T cells. PMID- 8417134 TI - Agonist regulation of gene expression of adrenergic receptors and G proteins. AB - Study of transmembrane signaling via G proteins has focused to a large extent upon investigations of individual G protein-linked receptor-effector systems. Agonist-induced desensitization and down-regulation of beta-adrenergic receptors, for example, have been studied extensively and adopted as a general model for G protein-linked receptor regulation. This review focuses not only on agonist regulation of adrenergic receptor gene expression, but also on how agonists regulate opposing adrenergic receptor-mediated pathways. This important feature of G protein-mediated pathways, i.e., cross-regulation and integration of information among several pathways, will be discussed in the context of what has been learned in the adrenergic receptor-coupled pathways. PMID- 8417135 TI - Rate of 59Fe uptake into brain and cerebrospinal fluid and the influence thereon of antibodies against the transferrin receptor. AB - Uptake of 59Fe from blood into brains of anaesthetized rats and mice has been studied by intravenous infusion of [59Fe]ferrous ascorbate or of 59Fe transferrin, the results not being significantly different. Uptakes in the rat were linear with time, but increased at longer times in the mouse. Transfer constants, K(in) (in ml/g/h x 10(3)), for cerebral hemispheres were 5.2 in the adult rat and 5.6 in the mouse. These K(in) values corresponded to 59Fe influxes of 145 and 322 pmol/g/h, respectively. 59Fe uptake into the mouse brain occurred in the following order: cerebellum > brainstem > frontal cerebral cortex > parietal cortex > occipital cortex > hippocampus > caudate nucleus. In genetically hypotransferrinaemic mice, 59Fe uptake into brain was 80-95 times greater than in To strain mice. Pretreatment of young rats and mice with monoclonal antibodies to transferrin receptors, i.e., the anti-rat immunoglobulin G OX 26 and the anti-mouse immunoglobulin M RI7 208, inhibited 59Fe uptake into spleen by 94% and 98%, respectively, indicating saturation of receptors. The antibodies reduced 59Fe uptake into rat brain by 35-60% and that into mouse brain by 65-85%. Although a major portion of iron transport across the blood-brain barrier is normally transferrin-mediated, non-transferrin-bound iron readily crosses it at low serum transferrin levels. PMID- 8417136 TI - Extracellular dopamine and serotonin in the rat striatum during transient ischaemia of different severities: a microdialysis study. AB - Generalised neurotransmitter overflow into the extracellular space on cerebral ischaemia has been widely reported and implicated in events leading to subsequent neuronal death. As little is known about the effect of depth of ischaemia on these changes, we have subjected anaesthetised rats to a sequence of four challenges [high K+ stimulus, moderate (penumbral) ischaemia, severe ischaemia, cardiac arrest] and have concurrently monitored both electrophysiological parameters and changes in extracellular dopamine, serotonin, and their metabolites in the striatum. Of particular relevance to human stroke therapy was penumbral ischaemia, where ionic homeostasis was maintained even though electrical function was lost. All challenges increased extracellular monoamines, although levels were significantly greater when ischaemia was severe enough to produce sustained anoxic depolarisation. Baseline levels were rapidly restored during recovery phases. Acidic monoamine metabolites decreased significantly during each insult, returning to basal levels during reperfusion after moderate ischaemia, and to significantly higher levels after severe ischaemia. Results indicate that sustained anoxic depolarisation may be a critical factor in determining outcome after ischaemia, being associated with significantly greater release of monoamines, and impairment of electrical function recovery. PMID- 8417137 TI - Effects of catechol-O-methyltransferase inhibitors and L-3,4 dihydroxyphenylalanine with or without carbidopa on extracellular dopamine in rat striatum. AB - The effects of two new catechol-O-methyltransferase (COMT) inhibitors, OR-611 and Ro 40-7592, in combination with L-3,4-dihydroxyphenylalanine (L-dopa) with or without carbidopa on extracellular levels of dopamine (DA), 3,4 dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), 3-O-methyldopa (3 OMD), and 5-hydroxyindoleacetic acid in rat striatum were studied. A dose of 10 mg/kg i.p. of Ro 40-7592 alone, in contrast to the same dose of OR-611, decreased the dialysate level of HVA and increased that of DOPAC; this dose was thus used to differentiate between the effects of central and peripheral COMT inhibition. L Dopa (50 mg/kg i.p.) alone slightly increased extracellular levels of DA, DOPAC, and HVA. The effects of L-dopa were potentiated by carbidopa (50 mg/kg i.p.), and even 3-OMD levels in dialysate samples became detectable. Both OR-611 and Ro 40 7592 significantly further increased the DA and DOPAC efflux from striatum produced by L-dopa. This increase was more pronounced when carbidopa was added to the treatment. OR-611 did not modify the effect of L-dopa or carbidopa/L-dopa on dialysate HVA levels, whereas Ro 40-7592 markedly reduced those levels. Both OR 611 and Ro 40-7592 very clearly suppressed dialysate 3-OMD levels produced by carbidopa/L-dopa. Ro 40-7592 was more effective than OR-611 in potentiating the effects of L-dopa or carbidopa/L-dopa. These in vivo data show that the new COMT inhibitors markedly inhibit the O-methylation of L-dopa and increase its availability to brain, which is reflected as increased DA formation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417138 TI - Mechanism of delayed increases in kynurenine pathway metabolism in damaged brain regions following transient cerebral ischemia. AB - Delayed increases in the levels of an endogenous N-methyl-D-aspartate receptor agonist, quinolinic acid (QUIN), have been demonstrated following transient ischemia in the gerbil and were postulated to be secondary to induction of indoleamine-2,3-dioxygenase (IDO) and other enzymes of the L-tryptophan kynurenine pathway. In the present study, proportional increases in IDO activity and QUIN concentrations were found 4 days after 10 min of cerebral ischemia, with both responses in hippocampus > striatum > cerebral cortex > thalamus. These increases paralleled the severity of local brain injury and inflammation. IDO activity and QUIN concentrations were unchanged in the cerebellum of postischemic gerbils, which is consistent with the preservation of blood flow and resultant absence of pathology in this region. Blood QUIN and L-kynurenine concentrations were not affected by ischemia. Brain tissue QUIN levels at 4 days postischemia exceeded blood concentrations, minimizing a role for breakdown of the blood-brain barrier. Marked increases in the activity of kynureninase, kynurenine 3 hydroxylase, and 3-hydroxyanthranilate-3,4-dioxygenase were also detected in hippocampus but not in cerebellum on day 4 of recirculation. In vivo synthesis of [13C6]QUIN was demonstrated, using mass spectrometry, in hippocampus but not in cerebellum of 4-day postischemic animals 1 h after intracisternal administration of L-[13C6]tryptophan. However, accumulation of QUIN was demonstrated in both cerebellum and hippocampus of control gerbils following an intracisternal injection of 3-hydroxyanthranilic acid, which verifies the availability of precursor to both regions when administered intracisternally. Notably, although IDO activity and QUIN concentrations were unchanged in the cerebellum of ischemic gerbils, both IDO activity and QUIN content were increased in cerebellum to approximately the same degree as in hippocampus, striatum, cerebral cortex, and thalamus 24 h after immune stimulation by systemic pokeweed mitogen administration, demonstrating that the cerebellum can increase IDO activity and QUIN content in response to immune activation. No changes in kynurenic acid concentrations in either hippocampus, cerebellum, or cerebrospinal fluid were observed in the postischemic gerbils compared with controls, in accordance with the unaffected activity of kynurenine aminotransferase activity. Collectively, these results support roles for IDO, kynureninase, kynurenine 3-hydroxylase, and 3-hydroxyanthranilate-3,4-dioxygenase in accelerating the conversion of L tryptophan and other substrates to QUIN in damaged brain regions following transient cerebral ischemia. Immunocytochemical results demonstrated the presence of macrophage infiltrates in hippocampus and other brain regions that parallel the extent of these biochemical changes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8417139 TI - Changes in the ganglioside long-chain base composition of rat cerebellar granule cells during differentiation and aging in culture. AB - Changes in the ganglioside long-chain base (LCB) composition in rat cerebellar granule cells in culture were studied during differentiation and aging. The total native ganglioside mixtures, extracted from the cells maintained in culture up to 22 days, were fractionated by reversed-phase HPLC, each ganglioside homogeneous in the oligosaccharide chain as well as in the LCB being quantified. Two main LCBs were components of the ganglioside species of cultured cells, the C18:1 LCB and the C20:1 LCB. The content of C20:1 ganglioside molecular species was low and quite constant during differentiation, comprising approximately 8% of the total ganglioside species content, the C20:1 LCB appearing to be represented more in the ganglioside of the "b series" (GD1b, GT1b, and GQ1b) than in the "a series" (GM1 and GD1a). During aging in culture, for 8-22 days, the content of the C20:1 species of all gangliosides increased, being more pronounced for GM1 and GD1a. PMID- 8417140 TI - Altered muscarinic and nicotinic receptor densities in cortical and subcortical brain regions in Parkinson's disease. AB - Muscarinic and nicotinic cholinergic receptors and choline acetyltransferase activity were studied in postmortem brain tissue from patients with histopathologically confirmed Parkinson's disease and matched control subjects. Using washed membrane homogenates from the frontal cortex, hippocampus, caudate nucleus, and putamen, saturation analysis of specific receptor binding was performed for the total number of muscarinic receptors with [3H]quinuclidinyl benzilate, for muscarinic M1 receptors with [3H]pirenzepine, for muscarinic M2 receptors with [3H]oxotremorine-M, and for nicotinic receptors with (-) [3H]nicotine. In comparison with control tissues, choline acetyl-transferase activity was reduced in the frontal cortex and hippocampus and unchanged in the caudate nucleus and putamen of parkinsonian patients. In Parkinson's disease the maximal binding site density for [3H]quinuclidinyl benzilate was increased in the frontal cortex and unaltered in the hippocampus, caudate nucleus, and putamen. Specific [3H]pirenzepine binding was increased in the frontal cortex, unaltered in the hippocampus, and decreased in the caudate nucleus and putamen. In parkinsonian patients Bmax values for specific [3H]oxotremorine-M binding were reduced in the cortex and unchanged in the hippocampus and striatum compared with controls. Maximal (-)-[3H]nicotine binding was reduced in both the cortex and hippocampus and unaltered in both the caudate nucleus and putamen. Alterations of the equilibrium dissociation constant were not observed for any ligand in any of the brain areas examined. The present results suggest that both the innominatocortical and the septohippocampal cholinergic systems degenerate in Parkinson's disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417141 TI - Kainate-induced changes in opioid peptide genes and AP-1 protein expression in the rat hippocampus. AB - In the rat hippocampus, jun, c-fos, and fos-related antigen immunoreactivity, AP 1 DNA binding, and opioid peptide gene expression were examined after kainate treatment to determine whether the induction and DNA binding of AP-1 transcription factors are correlated with the expression of the opioid peptide genes. One and one-half hours after kainate administration, fos-related antigen and jun immunoreactivity and AP-1 DNA binding were induced; maximal elevation was observed after 4.5 h. Transcription factor expression and DNA binding increased in a dose-dependent manner. Preprodynorphin and preproenkephalin mRNA induction was also dose dependent. The anticonvulsants, pentobarbital and diazepam, effectively blocked electroencephalographic seizure activity caused by kainate treatment, whereas valproic acid was approximately 50% effective. Opioid peptide gene expression, fos-related antigen and jun immunoreactivity, and AP-1 DNA binding all reflected similar reductions after anticonvulsant treatment. Therefore, expression and DNA binding activity of the AP-1 transcription factors are correlated with opioid peptide gene expression in the rat hippocampus. PMID- 8417142 TI - On the production and disposition of quinolinic acid in rat brain and liver slices. AB - The de novo production and subsequent disposition of the endogenous excitotoxin quinolinic acid (QUIN) was investigated in vitro in tissue slices from rat brain and liver. Incubation of tissue with QUIN's immediate bioprecursor 3 hydroxyanthranilic acid (3-HANA) in oxygenated Krebs-Ringer buffer yielded measurable amounts of QUIN both in the tissue and in the incubation medium. Saturation was reached between 16 and 64 microM 3-HANA (166 pmol of QUIN formed per milligram of protein after a 60-min incubation with 64 microM 3-HANA). In the brain, more QUIN was recovered from the tissue than from the incubation medium at all time points examined (5 min to 4 h). In contrast, the tissue-to-medium ratio for QUIN in parallel experiments with hepatic slices was << 1. The disposition of newly synthesized QUIN was further elaborated in tissue slices that had been preincubated for 60 min with 64 microM 3-HANA. Subsequent incubation of brain tissue in fresh buffer revealed a steady but relatively slow efflux of QUIN from the cellular compartment, with > 30% remaining in the tissue after a 90-min incubation. Analogous experiments with liver slices showed that > 93% of newly synthesized QUIN had entered the extracellular compartment within 30 min. Striatal and nigral slices obtained 7 days after an intrastriatal ibotenic acid injection showed severalfold increases in QUIN production compared with control tissues, in all likelihood due to astrogliosis and associated large increases in 3-hydroxyanthranilic acid oxygenase activity. In addition, the apparent tissue-to medium ratio was markedly reduced in striatal slices from lesioned animals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417143 TI - Production of adenosine from extracellular ATP at the striatal cholinergic synapse. AB - The components of the ectonucleotidase pathway at the immunoaffinity-purified striatal cholinergic synapse have been studied. The ecto-ATPase (EC 3.6.1.15) had a Km of 131 microM, whereas the ecto-ADPase (EC 3.6.1.6) had a Km of 58 microM, was Ca(2+)-dependent, and was inhibited by the ATP analogue 5' adenylylimidodiphosphate (AMPPNP). The ecto-5'-nucleotidase (EC 3.1.3.5) had a Km of 21 microM, was inhibited by AMPPNP and alpha,beta-methylene ADP, and by a specific antiserum. The Vmax values of the ATPase, ADPase, and 5'-nucleotidase enzymes present at this synapse were in a ratio of 30:14:1. Very little ecto adenylate kinase activity was detected on these purified synapses. The intraterminal 5'-nucleotidase enzyme, which amounted to 40% of the total 5' nucleotidase activity, was inhibited by AMPPNP, alpha,beta-methylene ADP, and the antiserum, and also had the same kinetic properties as the ectoenzyme. The time course of ATP degradation to adenosine outside the nerve terminals showed a delay, followed by a period of sustained adenosine production. The delay in adenosine production was proportional to the initial ATP concentration, was a consequence of feedforward inhibition of the ADPase and 5'-nucleotidase, and was inversely proportional to the ecto-5'-nucleotidase activity. The function and characteristics of this pathway and the central role of 5'-nucleotidase in the regulation of extraterminal adenosine concentrations are discussed. PMID- 8417144 TI - A brain-specific protein p25 is localized and associated with oligodendrocytes, neuropil, and fiber-like structures of the CA3 hippocampal region in the rat brain. AB - Developmental expression and cellular localization of a novel brain-specific 25 kDa protein (p25), a substrate of tau protein kinase II, were investigated in the rat brain using polyclonal antibodies raised against peptides synthesized based on the p25 amino acid sequence. By western immunoblotting, p25 was found to be expressed only slightly in the embryonic period; the expression increased from 11 days up to 5 weeks of age, and continued to increase gradually until 1-2 years of age. Immunohistochemistry revealed distinct staining of glial cells in most regions of the central nervous system in the adult rat brain. These positively immunostained cells were especially abundant in the white matter, such as the corpus callosum, cingulum, external capsule, and internal capsule. The glial cells were identified as oligodendrocytes, and the nuclei of the cells remained unstained. Whereas the neuropil in most parts of the brain was immunostained less intensely than glias, the neuropil in the first and second layers of the cerebral cortex and the dentate gyrus was relatively strongly stained. Fiber-like structures were also stained in the CA3 region of hippocampus. PMID- 8417145 TI - Endothelins stimulate c-fos and nerve growth factor expression in astrocytes and astrocytoma. AB - Endothelin receptors have been identified on astrocytes and astrocytoma, but their physiological significance has remained elusive. It is shown here that endothelins induce c-fos in primary cultures of mouse embryo astrocytes, as well as in two subclones of rat astrocytoma C6 cells, although with different kinetics. In addition, nerve growth factor expression is stimulated, as seen by mRNA accumulation and protein secretion, in primary astrocytes and one of the two C6 subclones, with an apparent correlation with the transience of c-fos induction. The activation of protein kinase C appears as an obligatory step during these processes, because (a) inhibition of protein kinase C by staurosporine blocks the induction by endothelin or phorbol esters of both c-fos and nerve growth factor, and (b) phorbol ester-evoked down-regulation of protein kinase C completely abolishes the c-fos induction by endothelin, but not that by the beta-adrenergic agonist isoproterenol, a known activator of the cyclic AMP dependent pathway. Our results support the hypothesis that c-fos product might be implicated in nerve growth factor expression by astrocytes, and also suggest that endothelins may participate in vivo in the modulation of the glial neurotrophic activity during brain development or wound healing. PMID- 8417146 TI - An improved solid-phase bioassay for evaluation of neurotrophic factors. AB - A bioassay system originally described by other investigators was improved to evaluate the biological activities of neurotrophic factors (NTFs) and is referred to as a solid-phase bioassay (SPB). The principle and an outline of the SPB are as follows: (a) Test samples containing NTFs are applied to polyacrylamide gel electrophoresis (PAGE) of the sodium dodecyl sulfate (SDS) or two-dimensional type and then transferred onto nitrocellulose membranes, (b) neurons are cultured on the protein-blotted membranes, and (c) the distributions of the surviving neurons are estimated following fixation and clarification of the nitrocellulose membranes. The rationale is that neuron survival will be restricted to the migration positions of the NTF(s). We used nerve growth factor (NGF) as a NTF and sympathetic neurons of 10- to 12-day-old chick embryos as NGF-responsive neurons. Neuronal survival was observed in the areas coinciding with the positions of NGF migration on the membranes in both systems following SDS-PAGE and two-dimensional PAGE. These results indicate that the SPB can identify the positions of NGF-like molecules on nitrocellulose membranes. Using this system, we found active entities, with molecular masses of approximately 100-200 kDa, different from NGF in crude extracts of mouse submaxillary glands. The SPB developed is considered to be a useful tool for obtaining information on the physicochemical and/or biological properties of putative NTFs in crude samples. PMID- 8417147 TI - Arachidonic acid and other unsaturated fatty acids alter membrane potential in PC12 and bovine adrenal chromaffin cells. AB - The action of arachidonic acid and other fatty acids on membrane potential in PC12 and bovine chromaffin cells was investigated using a membrane potential sensitive fluorescent dye. Arachidonic acid (1-40 microM) provoked dose-dependent membrane hyperpolarization, thereby reducing hyperpolarization induced by the K(+)-selective ionophore valinomycin. Other cis-unsaturated fatty acids, but not lipoxygenase products or the saturated fatty acid palmitic acid, also affected membrane potential. Tetraethylammonium blocked the arachidonic acid-induced hyperpolarization. These data suggest that cis-unsaturated fatty acids alter membrane potential in PC12 and bovine chromaffin cells by modulating K+ conductances. Valinomycin-generated hyperpolarization had no effect on agonist induced Ca2+ influx into bovine chromaffin cells, whereas preincubation with arachidonic acid and other cis-unsaturated fatty acids blocked Ca2+ influx and secretion. We propose a model where internally generated fatty acids act as a feedback to desensitize the stimulated cell via inhibition of receptor-dependent Ca2+ influx and induction of membrane hyperpolarization. PMID- 8417148 TI - Differential expression and subcellular localization of protein kinase C alpha, beta, gamma, delta, and epsilon isoforms in SH-SY5Y neuroblastoma cells: modifications during differentiation. AB - A decrease in protein kinase C activity caused either by treatment with inhibitors, such as staurosporine or H-7, or by prolonged exposure to phorbol diesters has been proposed to be involved in the early events of SH-SY5Y neuroblastoma cell differentiation. Because eight distinct isoforms of protein kinase C with discrete subcellular and tissue distributions have been described, we determined which isoforms are present in SH-SY5Y cells and studied their modifications during differentiation. The alpha, beta 1, delta, and epsilon isoforms were present in SH-SY5Y cells, as well as in rat brain. Protein kinase C alpha and -beta 1 were the most abundant isoforms in SH-SY5Y cells, and immunoreactive protein kinase C-delta and -epsilon were present in much smaller amounts than in rat brain. Subcellular fractionation and immunocytochemistry demonstrated that all four isoforms are distributed bimodally in the cytoplasm and the membranes. Immunocytochemical analysis showed that the alpha isoform is associated predominantly with the plasma membrane and the processes extended during treatment with 12-tetradecanoyl-13-acetyl-beta-phorbol or staurosporine, and that protein kinase C-epsilon is predominantly membrane-bound. Its localization did not change during differentiation. Western blots of total SH SY5Y cell extracts and of subcellular fractions probed with isoform-specific polyclonal antibodies showed that when SH-SY5Y cells acquired a morphologically differentiated phenotype, protein kinase C-alpha and -epsilon decreased, and protein kinase C-beta 1 did not change. These data suggest distinct roles for the different protein kinase C isoforms during neuronal differentiation, as well as possible involvement of protein kinase alpha and epsilon in neuritogenesis. PMID- 8417149 TI - Morphine inhibits calcium influx and the response to acetylcholine in Xenopus oocytes. AB - Incubation of intact Xenopus oocytes with the opioid radioligand [3H]diprenorphine (0.5 nM) resulted in specific binding of 1.7 +/- 0.3 fmol per oocyte. Morphine (10 microM) inhibited the uptake of 45Ca2+ into the oocyte by 66 +/- 9%. The opioid antagonist naltrexone partially blocked this effect of morphine. Preincubation of oocytes with morphine (10 microM, 2 min) partially inhibited the fast and slow responses of the oocyte to acetylcholine by 26 and 52%, respectively. We conclude that native Xenopus oocytes possess opioid receptors that may modulate the muscarinic response by limiting calcium influx into the cell. PMID- 8417150 TI - Type I calmodulin-sensitive adenylyl cyclase is neural specific. AB - The distribution of type I calmodulin-sensitive adenylyl cyclase in bovine and rat tissues was examined by northern blot analysis and in situ hybridization. Northern blot analysis using poly(A)(+)-selected RNA from various bovine tissues indicated that mRNA for type I adenylyl cyclase was found only in brain, retina, and adrenal medulla, suggesting that this enzyme is neural specific. In situ hybridization studies using bovine, rabbit, and rat retina indicated that mRNA for type I adenylyl cyclase is found in all three nuclear layers of the neural retina and is particularly abundant in the inner segment of the photoreceptor cells. The neural-specific distribution of type I adenylyl cyclase mRNA and its restricted expression in areas of brain implicated in neuroplasticity are consistent with the proposal that this enzyme plays an important role in various neuronal functions including learning and memory. PMID- 8417151 TI - Neuroblastoma growth factors derived from neurofibroma (NF1): participation of uridine in a neuroblastoma growth. AB - Human glioma cell extracts were found to elicit a marked growth-promoting activity on human neuroblastoma cells. This activity was also detected in the extracts of neurofibroma type 1 (NF1; von Recklinghausen neurofibromatosis) comprising aberrant Schwann cell growth. The purified substance from the NF1 extracts by HPLC on ODS columns was identical to a pyrimidine nucleoside, uridine, the chemical structure of which was identified by gas chromatography mass spectrometry. The authentic uridine showed a strong growth-promoting activity on human neuroblastoma cells. Other purine or pyrimidine nucleotides, their derivatives, and ribose sources for their syntheses were employed to test the activity; a purine nucleoside, adenosine, showed a stronger activity than uridine. The current study raises the possibility that human neuroblastoma cells may be affected by dysfunctions of the de novo pathway of both purine and pyrimidine nucleotide biosyntheses. PMID- 8417152 TI - Characterization of a triacylglycerol lipase that liberates arachidonic acid from bovine chromaffin cells during secretion. AB - Primary cultures of chromaffin cells from bovine adrenal medullae were used as a model to study lipolytic events during stimulus-secretion coupling. It has been shown that chromaffin cells liberate arachidonic acid in addition to their main secretion product, the catecholamines. To understand more about the mechanism of arachidonic acid liberation, chromaffin cells were labeled with radioactive arachidonic acid, stimulated, and then analyzed for changes in lipid composition. After stimulation with 10(-4) M acetylcholine, the radioactivity of triacylglycerols decreased to the same extent that the free arachidonic acid level rose. This finding suggests that in bovine chromaffin cells a stimulation dependent triacylglycerol lipase (triacylglycerol hydrolase; EC 3.1.1.3) is involved in arachidonic acid liberation. Further work was performed on detection, characterization, and isolation of this enzyme. Triacylglycerol lipase activity was found in whole cell homogenates and in plasma membrane fractions isolated from adrenal medullary tissue. The plasma membrane lipase showed a pH optimum of 4.3. The apparent Michaelis constant was determined as 3.3 x 10(-4) mol/L. Ca2+ did not influence the enzymatic activity. To differentiate the plasma membrane triacylglycerol lipase from the previously described plasma membrane diacylglycerol lipase of chromaffin cells, the influence of RG 80267, a specific diacylglycerol lipase inhibitor, was examined. RG 80267 (50 microM) inhibited the triacylglycerol lipase by only 24%, although diacylglycerol lipase was totally inhibited with only 20 microM RG 80267. The pH optimum of homogenate lipase was broad, lying between 4 and 7. Starting from the soluble fraction of whole cell homogenates, the triacylglycerol lipase was partially purified by ultracentrifugation and size-exclusion chromatography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417153 TI - Basis for phospholipid incorporation into peripheral nerve myelin. AB - To characterize the mechanism(s) for targeting of phospholipids to peripheral nerve myelin, we examined the kinetics of incorporation of tritiated choline-, glycerol-, and ethanolamine-labeled phospholipids into four subfractions: microsomes, mitochondria, myelin-like material, and purified myelin at 1, 6, and 24 h after precursors were injected into sciatic nerves of 23-24-day-old rats. As validation of the fractionation scheme, a lag (> 1 h) in the accumulation of labeled phospholipids in the myelin-containing subfractions was found. This lag signifies the time between synthesis on organelles in Schwann cell cytoplasm and transport to myelin. In the present study, we find that sphingomyelin (choline labeled) accumulated in myelin-rich subfractions only at 6 and 24 h, whereas phosphatidylserine (glycerol-labeled) and plasmalogen (ethanolamine-labeled) accumulated in the myelin-rich fractions by 1 h. The later phospholipids accumulate preferentially in the myelin-like fraction. These results are consistent with the notion that the targeting of sphingomyelin, a lipid present in the outer myelin leaflet, is different from the targeting of phosphatidylserine and ethanolamine plasmalogen, lipids in the inner leaflet. These findings are discussed in light of the possibility that sphingomyelin targeting is Golgi apparatus based, whereas phosphatidylserine and ethanolamine plasmalogen use a more direct transport system. Furthermore, the routes of phospholipid targeting mimic routes taken by myelin proteins P0 (Golgi) and myelin basic proteins (more direct). PMID- 8417154 TI - Identification of p42 mitogen-activated protein kinase as a tyrosine kinase substrate activated by maximal electroconvulsive shock in hippocampus. AB - Recent studies have demonstrated that administration of an electroconvulsive shock produces a rapid and transient increase in tyrosyl phosphorylation of a approximately 40-kDa protein in rat brain. Initial characterization of this protein's chromatographic properties indicated that it might be a member of a recently identified family of kinases, referred to as mitogen-activated protein (MAP) kinases, that are activated by tyrosyl phosphorylation. In the present study, we have used MAP kinase antisera to assess the identity of this protein. We have found that the approximately 40-kDa phosphotyrosine-containing protein comigrates with p42 MAP kinase (p42mapk) and not with two other 44-kDa MAP kinase family members detected by these antisera. Western blots of proteins immunoprecipitated with MAP kinase antibodies confirm that p42mapk displays increased tyrosyl phosphorylation after an electroconvulsive stimulus. Chromatographic separation of hippocampal extracts indicates that MAP kinase activity elutes in parallel with p42mapk. Accordingly, these studies identify p42mapk as a tyrosyl kinase substrate that is activated by this stimulus and suggest that this form of MAP kinase may be selectively regulated by neuronal stimulation. PMID- 8417155 TI - Cloning and expression of a rat acetylcholinesterase subunit: generation of multiple molecular forms and complementarity with a Torpedo collagenic subunit. AB - We obtained a cDNA clone encoding one type of catalytic subunit of acetylcholinesterase (AChE) from rat brain (T subunit). The coding sequence shows a high frequency of (G+C) at the third position of the codons (66%), as already noted for several AChEs, in contrast with mammalian butyrylcholinesterase. The predicted primary sequence of rat AChE presents only 11 amino acid differences, including one in the signal peptide, from that of the mouse T subunit. In particular, four alanines in the mouse sequence are replaced by serine or threonine. In northern blots, a rat AChE probe indicates the presence of major 3.2- and 2.4-kb mRNAs, expressed in the CNS as well as in some peripheral tissues, including muscle and spleen. In vivo, we found that the proportions of G1, G2, and G4 forms are highly variable in different brain areas. We did not observe any glycolipid-anchored G2 form, which would be derived from an H subunit. We expressed the cloned rat AChE in COS cells: The transfected cells produce principally an amphiphilic G1a form, together with amphiphilic G2a and G4a forms, and a nonamphiphilic G4na form. The amphiphilic G1a and G2a forms correspond to type II forms, which are predominant in muscle and brain of higher vertebrates. The cells also release G4na, G2a, and G1a in the culture medium. These experiments show that all the forms observed in the CNS in vivo may be obtained from the T subunit. By co-transfecting COS cells with the rat T subunit and the Torpedo collagenic subunit, we obtained chimeric collagen-tailed forms. This cross-species complementarity demonstrates that the interaction domains of the catalytic and structural subunits are highly conserved during evolution. PMID- 8417156 TI - N-methyl-D-aspartate receptor excitotoxicity involves activation of polyamine synthesis: protection by alpha-difluoromethylornithine. AB - We investigated the role of polyamines and their regulatory enzyme ornithine decarboxylase in N-methyl-D-aspartate-induced excitotoxicity in embryonic chick retina. N-Methyl-D-aspartate (200 microM) produced an early increase in ornithine decarboxylase activity, putrescine concentration, and Ca2+ entry, leading to selective neuronal death by 30 min. This response was attenuated by the ornithine decarboxylase inhibitor alpha-difluoromethylornithine and the N-methyl-D aspartate receptor antagonist 5-aminophosphonovaleric acid. Exogenous putrescine increased intracellular putrescine and spermine levels and reversed neuroprotection by alpha-difluoromethylornithine, but not by 5 aminophosphonovaleric acid. N-Methyl-D-aspartate-receptor stimulation of putrescine/polyamine synthesis mediates abnormal Ca2+ entry and acute excitotoxic neuronal death. Postreceptor inhibition of the ornithine decarboxylase/polyamine cascade by alpha-difluoromethylornithine may provide neuroprotection against N methyl-D-aspartate-induced excitotoxicity. PMID- 8417157 TI - Age-dependent vulnerability of the striatum to the mitochondrial toxin 3 nitropropionic acid. AB - The mechanisms of delayed onset and cell death in Huntington's disease (HD) are unknown. One possibility is that a genetic defect in energy metabolism may result in slow excitotoxic neuronal death. Therefore, we examined the effects of age on striatal lesions produced by local administration of the mitochondrial toxin 3 nitropropionic acid in rats. In vivo chemical shift magnetic resonance imaging showed marked increases in striatal lactate concentrations that significantly correlated with increasing age. Histologic and neurochemical studies showed a striking age dependence of the lesions, with 4- and 12-month-old animals being much more susceptible than 1-month-old animals. Continuous systemic administration of low doses of 3-nitropropionic acid for 1 month resulted in striatal lesions showing growth-related changes in dendrites of striatal spiny neurons using the Golgi technique. These results show that a known mitochondrial toxin can produce selective axon-sparing striatal lesions showing both the age dependence and striatal spiny neuron dendritic changes that characterize HD. PMID- 8417158 TI - Inhibition of protein synthesis during repetitive cortical spreading depression. AB - The effect of cortical spreading depression (CSD) on cerebral protein synthesis (CPS) was examined. CSD was evoked in normal rats with KCl, and CPS was measured autoradiographically with [1-14C]leucine. Average rates (mean +/- SD) of CPS in layers I-IV of cortex decreased significantly from 10.7 +/- 0.6 (sham-operated controls; n = 4) to 6.7 +/- 0.7 nmol/g/min (n = 4; p < 0.01) and in layers V-VI from 10.9 +/- 0.5 to 9.4 +/- 0.4 nmol/g/min (p < 0.05) during 60 min of repetitive CSD. Spreading depression did not affect CPS rates in other subcortical brain regions. These results indicate that KCl-evoked CSD induces inhibition but not suppression of cortical protein synthesis. PMID- 8417159 TI - ADP-ribosylation of the neuronal phosphoprotein B-50/GAP-43. AB - The neuronal phosphoprotein B-50/GAP-43 is associated with growth and regeneration within the nervous system and its posttranslational status can be correlated with its cellular localization during growth and regeneration. Recently, B-50 has been shown to interact with certain G protein subunits. Regulation of G protein-mediated signal transduction may involve ADP-ribosylation in vivo. In the present study we have demonstrated that B-50 is a substrate for endogenous ADP-ribosyltransferases. The results are discussed with respect to the possible interaction of B-50 with G proteins, but also with regard to the posttranslational modification of B-50 by all major regulatory mechanisms that act at, or through, the neuronal membrane. PMID- 8417160 TI - Differences in the ligand binding properties of the short and long versions of the D2 dopamine receptor. AB - The short and long forms of the D2 dopamine receptor have been expressed at similar levels in fibroblast Ltk- cells, and the long form of the receptor has also been expressed in Chinese hamster ovary cells. The ligand binding properties of the two forms of receptor expressed in different systems have been compared in saturation analyses with [3H]spiperone and in competition studies with a range of antagonists. The long form of the receptor expressed in two different cell hosts exhibits essentially identical properties. However, whereas the long and short forms of the receptor show very similar affinities for several compounds representative of different chemical classes, the short form shows a two- to fivefold higher affinity for several substituted benzamide drugs. The conformation of the receptor binding site may therefore be different in the two receptor isoforms. PMID- 8417161 TI - Co-administration of the D2 antagonist pimozide inhibits up-regulation of dopamine release and uptake induced by repeated cocaine. AB - To investigate the hypothesis that the D2 dopamine (DA) receptor regulates DA uptake, as well as release, in the nucleus accumbens (N ACC), rats were pretreated for 10 days with either the selective D2 antagonist pimozide (1.0 mg/kg, i.p.) or vehicle, followed 3 h later by either cocaine (20 mg/kg, i.p.) or saline. On day 11, a microdialysis method was performed in which various DA concentrations (0, 10, and 20 nM DA) were perfused through the dialysis probe to characterize the diffusion of DA through tissue to and from the microdialysis probe (recovery). This diffusion of DA has been shown to be sensitive to changes in release and uptake. Pimozide pretreatment was shown to attenuate significantly a cocaine-induced increase in the in vivo recovery of DA (p < 0.01). The in vivo recovery for the vehicle/cocaine group was 47 +/- 4%, whereas the in vivo recovery for the pimozide/cocaine group was 31 +/- 3%. There was no difference between the pimozide/cocaine and control groups (pimozide/saline, 26 +/- 2%; vehicle/saline, 26 +/- 3%). In vitro probe calibrations indicated no significant difference in probe efficiencies between groups. These data suggest that the D2 receptor is capable of modulating uptake as well as release of DA in the N ACC of the rat. PMID- 8417162 TI - A single amino acid difference accounts for the pharmacological distinctions between the rat and human 5-hydroxytryptamine1B receptors. AB - Molecular cloning of the rat and human 5-hydroxytryptamine1B (5-HT1B) receptors has revealed that the primary amino acid sequence of these two receptors is > 90% identical. Despite this high degree of primary sequence homology, these two receptors have significantly different pharmacological properties. A mutant human 5-HT1B receptor was constructed in which Thr355 was replaced by Asn, the corresponding residue at this position in the rat 5-HT1B receptor. The pharmacology of the mutant human 5-HT1B receptor was very similar to that of the rat 5-HT1B receptor. Specifically, the mutant receptor had much higher affinity for pindolol, [125I]-iodocyanopindolol, propranolol, and CP-93,129 than the wild type receptor. In contrast, the mutant had significantly lower affinity for sumatriptan, N,N-dipropyl-5-carboxamidotryptamine, 5-methoxy-N,N dimethyltryptamine, methysergide, metergoline, and rauwolscine. These data suggest that a single amino acid difference at position 355 is responsible for the pharmacological differences between the rat and human 5-HT1B receptors. PMID- 8417163 TI - A molecular mechanism of aluminum neurotoxicity. PMID- 8417164 TI - In memoriam: Henry McIlwain 1912-1992. PMID- 8417165 TI - Pre- and posttranslational regulation of beta-endorphin biosynthesis in the CNS: effects of chronic naltrexone treatment. AB - There appear to be two anatomically distinct beta-endorphin (beta E) pathways in the brain, the major one originating in the arcuate nucleus of the hypothalamus and a smaller one in the area of the nucleus tractus solitarius (NTS) of the caudal medulla. Previous studies have shown that these two proopiomelanocortin (POMC) systems may be differentially regulated by chronic morphine treatment, with arcuate cells down-regulated and NTS cells unaffected. In the present experiments, we examined the effects of chronic opiate antagonist treatment on beta E biosynthesis across different CNS regions to assess whether the arcuate POMC system would be regulated in the opposite direction to that seen after opiate agonist treatment and to determine whether different beta E-containing areas might be differentially regulated. Male adult rats were administered naltrexone (NTX) by various routes for 8 days (subcutaneous pellets, osmotic minipumps, or repeated intraperitoneal injections). Brain and spinal cord regions were assayed for total beta E-ir, different molecular weight immunoreactive beta endorphin (beta E-ir) peptides, and POMC mRNA. Chronic NTX treatment, regardless of the route of administration, reduced total beta E-ir concentrations by 30-40% in diencephalic areas (the arcuate nucleus, the remaining hypothalamus, and the thalamus) and the midbrain, but had no effect on beta E-ir in the NTS or any region of the spinal cord. At the same time, NTX pelleting increased POMC mRNA levels in the arcuate to approximately 140% of control values.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417166 TI - Transport of phosphatidylserine from microsomes to the inner mitochondrial membrane in brain tissue. AB - Phosphatidylserine was labeled by incubating rat brain homogenates with [3 14C]serine in the presence of Ca2+ (base-exchange conditions). Some labeled phosphatidylethanolamine also forms, in spite of the inhibition of Ca2+ on phosphatidylserine decarboxylase. Phosphatidylserine labeling and decarboxylation also occur on incubating a mixture of purified mitochondria and microsomes, suggesting that no soluble factors are necessary for the synthesis and the decarboxylation of phosphatidylserine. Ca2+ favors the transfer of phosphatidylserine from microsomes (where it forms) to mitochondria (where it is decarboxylated). The specific radioactivity of the phosphatidylserine transferred to mitochondria is higher than that of microsomal phosphatidylserine. This finding supports the hypothesis that the lipid is compartmentalized in microsomes and that radioactive, newly synthesized phosphatidylserine is much better exported than the bulk of microsomal phospholipid. PMID- 8417167 TI - Ischemia-induced accumulation of extracellular amino acids in cerebral cortex, white matter, and cerebrospinal fluid. AB - In a global model of brain ischemia, accumulation of amino acids was studied in the extracellular space of the auditory cortex and the internal capsule using microdialysis, and in CSF of halothane anesthetized cats. In both brain regions, blood flow determined by hydrogen clearance decreased below 10 ml/100 g/min after extracranial multiple-vessel occlusion, and extracellular potassium activity (Ke) measured in the dialysate increased significantly. A delayed rise in Ke was observed in CSF. In contrast, ischemic amino acid accumulation differed markedly between the two brain regions investigated. In cortex, transmitter amino acids glutamate, aspartate, and gamma-aminobutyric acid (GABA) rose almost immediately after onset of ischemia, and increased 30-, 25-, and 250-fold, respectively, after 2 h of ischemia. The nontransmitter amino acids taurine, alanine, and serine increased 10-, seven-, and fourfold, respectively, whereas glutamine and essential amino acids (valine, phenylalanine, isoleucine, and leucine) increased only 1.5-fold. In the internal capsule, increases in amino acids, if any, were delayed and much smaller than in cortex. The largest alteration was a fivefold elevation of GABA. In CSF, changes in amino acids were small and comparable to those in the internal capsule. Our results demonstrate that ischemia-induced extracellular amino acid accumulation is a well localized phenomenon restricted to gray matter structures that possess release and reuptake systems for these substances. We assume that amino acids diffuse slowly into adjacent while matter structures, and into CSF. PMID- 8417168 TI - Expression of gangliosides GD3 and 3'-isoLM1 in autopsy brains from patients with malignant tumors. AB - Three autopsy brains from patients who succumbed to malignant gliomas have been analyzed in various regions with regard to their ganglioside content. The study focused on the gangliosides GD3 and 3'-isoLM1, which in a previous study of biopsies were found to be associated with these tumors. In particular, 3'-isoLM1, was suggested to be a marker for malignant gliomas. The highest concentrations (200-1,000 nmol of sialic acid/g wet weight) of GD3 was found in specimens of macroscopically pure tumor, where the proportion of GD3 was, at the most, 78% (range, 11-78%) of the total ganglioside sialic acid compared with < 10% in normal brain tissue. The proportion of the total ganglioside sialic acid made up by GD3 was also elevated in the periphery of the tumor and in the same region in the opposite hemisphere, where no tumor cells were detected. In four of eight brain metastases of various carcinomas, GD3 was > 10% of the total ganglioside sialic acid (range, 3-37%). The ganglioside 3'-isoLM1, as determined by TLC enzyme-linked immunosorbent assay using a specific monoclonal antibody (SL-50), was not present at detectable levels in any of the macroscopically homogenous tumor areas. It was, however, found in the periphery of the tumor, in the corpus callosum, and at highest concentrations in the region of the opposite hemisphere corresponding to the tumor. The concentration varied between 0.1 and 6.0 nmol/g wet weight of tissue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417169 TI - Lack of evidence that human immunodeficiency virus can infect human endothelial cells in vitro. PMID- 8417170 TI - Acquired protein S deficiency. PMID- 8417171 TI - Transfer and metabolism of dideoxyinosine by the perfused human placenta. AB - Dideoxyinosine (DDI) has been recently approved for the treatment of AIDS. In anticipation of its use in HIV-infected women during pregnancy, the transfer and metabolism of DDI by the perfused human placenta have been investigated. Transfer characteristics are those of simple diffusion: clearance is the same as that for L-glucose (transfer index of 0.98 +/- 0.09), it is equivalent in both directions across the placenta, and the transfer rate is proportional to the transplacental gradient over a very broad range (1 to 500 microM). Because of extensive placental metabolism, only about one-half of the cleared DDI (51 +/- 21%) is transferred intact to the fetal circulation. No dideoxyadenine triphosphate, the antiviral product of DDI, could be detected in the placenta following perfusion. Comparison of the pharmacological information on DDI and zidovudine (ZDV) indicates that treatment of HIV-infected women during pregnancy with DDI will expose the fetus to much less drug than if ZDV were used. DDI may therefore be less effective than ZDV in the treatment of the infected fetus. However, the uninfected fetus of an HIV-infected woman will gain by reduced exposure to a drug that is known to be toxic. PMID- 8417172 TI - Multicenter clinical trial of oral ribavirin in symptomatic HIV-infected patients. The Ribavirin ARC Study Group. AB - A double-blind, randomized, placebo-controlled trial comparing two daily doses of oral ribavirin (600 mg and 800 mg) and placebo was conducted at six medical centers. Two hundred fifteen adults were enrolled over a 5-month period and randomized to receive ribavirin 800 mg daily (74 subjects), ribavirin 600 mg (71 subjects), or placebo (70 subjects). Active treatment was administered for 24 weeks followed by a 4-week wash-out period. Fifteen patients receiving placebo, 10 receiving ribavirin 600 mg, and 18 receiving ribavirin 800 mg developed AIDS during the 28-week study period. Ribavirin at daily doses of 600 mg or 800 mg for 24 weeks did not significantly affect the rate of progression to AIDS as defined by the Centers for Disease Control, in univariate analysis, Kaplan-Meier survival analysis, or Cox proportional hazards modeling. Although in the proportional hazards analysis, the dose-response regression coefficients indicated a reduction of 43% in the hazard of progression to AIDS among patients on active treatment, it was not statistically significant (p = 0.19). Furthermore, there was no evidence that ribavirin had a significant effect upon any immunologic or virologic parameter measured, including CD4 count, CD4:CD8 ratio, lymphocyte proliferative response, skin test reactivity, interferon-gamma production, peripheral blood mononuclear cell culture for human immunodeficiency virus (HIV), or level of HIV p24 antigen in serum. The most prominent adverse effect of ribavirin was induction of a mild reversible hemolytic anemia (mean fall in hematocrit, 5%). No severe or unremitting drug reaction was documented. PMID- 8417173 TI - Reduced frequency of HIV-induced brain lesions in AIDS patients treated with zidovudine. AB - We evaluated the effect of zidovudine on HIV-induced lesions of the brain by comparing the neuropathological findings in 82 treated and 120 untreated patients who died from AIDS. We observed a statistically significant reduction of the number of cases with multinucleated giant cells (MGCs) in the brain and MGC associated neuropathological damage in patients treated with zidovudine. The effects of zidovudine were time and dose related in the first 12 months of treatment, while longer periods of therapy produced no further results. The antiretroviral treatment particularly affected the frequency of diffuse demyelinating lesions of the cerebral white matter. In the patients who died with HIV-induced brain lesions but no other opportunistic brain diseases, the percentage of cases with clinical history of severe dementia was significantly lower in the group treated with zidovudine. PMID- 8417174 TI - Recommendations for prophylaxis against Pneumocystis carinii pneumonia for persons infected with human immunodeficiency virus. U.S. Public Health Service Task Force on Antipneumocystis Prophylaxis in Patients with Human Immunodeficiency Virus Infection. PMID- 8417175 TI - Pharmacokinetics of zidovudine alone and in combination with oxazepam in the HIV infected patient. AB - This three-phase study was designed to determine if a pharmacokinetic drug-drug interaction exists between zidovudine and oxazepam. Six individuals infected with human immunodeficiency virus (HIV) and receiving zidovudine at 500 mg daily, with normal renal and hepatic function, were enrolled. During phase I, zidovudine pharmacokinetics were studied after steady-state oral administration (100 mg every 4 h) and after a single dose (70 mg) of intravenous zidovudine. Phase II consisted of a single oral dose (30 mg) of oxazepam followed by a 48-h blood sampling period. Phase III began with 48 h of concomitant zidovudine, 100 mg orally every 4 h, and oxazepam, 15 mg orally every 8 h, followed by concomitant dosing of intravenous zidovudine and oral oxazepam. Zidovudine concentrations were determined by radioimmunoassay. Oxazepam concentrations were determined with use of a fluorescence polarization immunoassay. The calculated bioavailability was 0.61 for zidovudine alone and 0.75 when administered in combination with oxazepam (p = 0.16). Plasma half-life for oral zidovudine alone and in combination with oxazepam was 1.17 h versus 0.99 h, respectively (p = 0.25), and 1.38 h versus 1.15 h (p = 0.38) for intravenous zidovudine during single and combination therapy, respectively. Total body clearance of zidovudine was not significantly altered by oxazepam (93 L/h vs. 109 L/h, p = 0.16). The mean pharmacokinetic parameters determined for a single 30-mg dose of oxazepam for oral clearance, apparent volume of distribution, and plasma half-life were 9.8 L/h, 65.7 L, and 5.1 h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417176 TI - Signs and symptoms of "asymptomatic" HIV-1 infection in homosexual men. Multicenter AIDS Cohort Study. AB - We investigated the long-term health effects of HIV-1 infection in homosexual men not close to developing AIDS by comparing 916 HIV-1-seropositive (SP) men at least 1.67-3.67 years prior to a clinical AIDS diagnosis to 2,161 HIV-1 seronegative (SN) controls. The SP group reported a higher total of 12 distinct symptoms (fatigue, shortness of breath, night sweats, rash, cough, diarrhea, headache, thrush, skin discoloration, fever, weight loss, and sore throat/mouth) than did the SN group (p < 0.0001), corresponding to at least 5.6 more days/year of such symptoms. The SP group had lower body mass index (p < 0.0001) and lower hemoglobin (p < 0.0001). The SP group was more depressed, as measured by CES-D score (p = 0.047), before knowledge of one's serostatus was likely, and became even further depressed (p = 0.038 for increase in depression) after the HIV-1 serostatus test was accessible to high-risk groups. These associations remained unchanged in multivariate models, incorporating other covariates. PMID- 8417177 TI - Neutralization of HIV-1 by F105, a human monoclonal antibody to the CD4 binding site of gp120. AB - The functional ability of the human monoclonal antibody (HMab) F105 to neutralize commonly available laboratory strains and a selection of primary isolates of human immunodeficiency virus (HIV)-1 was studied. F105 is representative of a class of human antibodies that react with conformational epitopes within the discontinuous CD4 binding site on HIV-1 envelope glycoprotein gp120. F105 binds with relatively similar affinities to native antigen expressed on the surfaces of cells infected with each of five laboratory isolates tested (IIIB, SF2, MN, RF, and CC) and neutralizes SF2, IIIB, and MN with concentrations of antibody ranging from 140 ng to 10 micrograms/ml. Nonetheless, neutralization by F105 alone of RF and CC is poor at modest antibody concentrations despite high affinity binding to surface gp120 on infected cells. Neutralization of HIV-1 strains by F105 is unaffected by normal sera and cooperativity is observed with serum samples from HIV-1 infected patients. Of significance, neutralization of RF and MN by F105 is enhanced by some HIV-seropositive sera at low concentrations. F105 also neutralized 30% of HIV-1 primary isolates in lymphocyte cultures. Although it is unclear how relevant in vitro studies will be to in vivo events, these data allow comparison of F105 with other HMabs to the CD4 binding site and V3 loop and provide an in vitro reference for in vivo activity. These studies demonstrate that antibody interactions among different classes of antibodies may be important in in vivo neutralization of HIV-1. PMID- 8417178 TI - Virus excretion in the cervicovaginal secretions of pregnant and nonpregnant HIV infected women. AB - In order to estimate the prevalence of viral excretion in cervicovaginal secretions, we made a cross-sectional study of 55 HIV-infected women. The patient population was diverse, including pregnant and nonpregnant women in different disease stages from three centers. Virus replication was found in the cell-free supernatant from 12 of 55 cervicovaginal samples (21.8%) by coculture on the CD4 positive cell line CEM-C113. In addition, cell-associated virus was detected in five of a subgroup of 22 samples testing negatively on cell-free supernatant. The prevalence of HIV in the cell-free supernatant was not related to disease stage, zidovudine therapy, transmission group, or history of sexually transmitted diseases. Excretion of HIV was significantly higher in our population of pregnant women (eight of 21, 38%) compared with an unmatched group of nonpregnant women (four of 34, 11.8%; p = 0.04). These results provide evidence of cell-free virus shedding as well as the presence of cell-associated virus in the genital secretions of HIV-infected women. PMID- 8417179 TI - Using indices to differentiate dimensions of knowledge regarding modes of HIV transmission in the U.S. population, 1987-1989. AB - The number of HIV-infected individuals is increasing, making it important for the public to understand not only how HIV is transmitted but also the lack of transmission risk associated with casual contact. Using CDC's National Center for Health Statistics National Health Interview Survey, we divided modes of transmission items into two areas of knowledge: "True Transmission" and "False Transmission." Items were recoded with scores from 3 for the most correct response to 0 for the most incorrect response for each of three items related to true and each of eight items related to false transmission. Item and principal components factor analyses yielded two distinct dimensions (true factor loadings from 0.68 to 0.76, false factor loadings from 0.56 to 0.74). Mean scores of 8.3 (range 0-9) and 15.9 for 1987 (range 0-24) for true and false transmission indices, respectively, provide evidence that the population is highly knowledgeable about true modes of transmission but far less so about false modes. Knowledge levels have increased between 1987 and 1989, most meaningfully in the area of false transmission. Use of these indices will facilitate the monitoring over time of differential knowledge, attitudes, and beliefs related to HIV and AIDS. PMID- 8417180 TI - Trends in self-reported HIV risk behavior: injection drug users in Los Angeles. AB - This article reviews trends in HIV risk behaviors across serial samples of Los Angeles injection drug users interviewed between 1987 and 1991. All indicators are based on self-reported behavior during the year before the interview. Findings show persistent drug-related risk behaviors. No decrease has occurred in needle sharing with strangers/acquaintances or at shooting galleries. No increase has occurred in self-reported avoidance of needle sharing for as long as 1 year. The only exception to this pattern is a significant increase in bleach use among injection drug users who share needles. Findings are mixed regarding sex-related risk behaviors. Drug users have not reduced their yearly number of sex partners, but condom use has become more prevalent among nonmonogamous drug users. We conclude that preventive education will need to adopt new strategies for addressing persistent risk behaviors while at the same time reinforcing favorable trends that have already begun. PMID- 8417181 TI - Effects of increasing outpatient diagnosis on AIDS surveillance. AB - Using AIDS surveillance data, we analyzed trends and correlates of outpatients AIDS diagnosis in Oregon and Washington. The proportion of outpatient diagnoses rose from 24% of cases in 1987 to 51% in 1990. Case characteristics associated with outpatient diagnosis included white race, urban residence, and the exposure category of male homosexual/bisexual contact. AIDS-defining conditions associated with outpatient diagnosis included Kaposi's sarcoma, HIV wasting syndrome, and esophageal candidiasis. Completeness and timeliness of reporting was poorer for cases diagnosed as outpatients compared with inpatients. As outpatient diagnosis becomes more common, modified surveillance methods may be needed to ensure complete case finding and consequent reliability of AIDS surveillance information. PMID- 8417182 TI - Screening for HIV and hepatitis B virus in Los Angeles County prenatal clinics: a demonstration project. AB - A project designed to pilot voluntary screening for HIV antibody and hepatitis B virus antigen (HBsAg) in women using Los Angeles County Department of Health Services prenatal clinics is described. The purpose of the study was to demonstrate that HIV testing could be integrated into an existing prenatal health care system with minimal disruption. In an 8-month period, 9,069 women entered the project; 76% accepted HIV antibody testing. The rate of HIV antibody seropositivity was 144 per 100,000 (10 per 6,943), and the rate of HBsAg seropositivity was 253 per 100,000 (23 per 9,103). No difference in test acceptance rates was found using a sign-off versus a sign-on HIV antibody consent form although potential confounders were not controlled. Only five of the eight HIV antibody-positive women (63%) and eight of the 20 HBsAg-positive women (40%) who were interviewed reported behaviors considered high risk for HIV or HBsAg infection. PMID- 8417183 TI - Seroepidemiology of human immunodeficiency virus infection in Borno State of Nigeria by sentinel surveillance. AB - A serosurvey for human immunodeficiency virus (HIV) infection was carried out in three well-separated population centers in Borno State of Nigeria in 1989-1990. The study subjects were 1,259 made up of sexually transmitted disease (STD) clinic attenders, pregnant women, female prostitutes, and blood donors. Sera were screened by enzyme-linked immunosorbent assay and confirmation was done by Western blotting. The overall seroprevalence was 1.67% (21/1,259), with no significant differences from one population center to another. There were, however, significant differences among the population groups studied: prostitutes, 4.24%; STD clinic attenders, 1.67%; blood donors, 0.71%; and pregnant women, 0.24%. Of the 21 seropositives, 18 were positive for HIV-1 only; 1 was positive for HIV-2 only; and 2 were positive for both HIV-1 and HIV-2. All three HIV-2-positive sera were from prostitutes. Prevalence rates found in this study showed marked increases from 2 to 3 years earlier. There is therefore the need for a vigorous and sustained intervention program. PMID- 8417184 TI - Nephron sparing surgery for renal cell carcinoma. PMID- 8417185 TI - Successful management of bilateral xanthogranulomatous pyelonephritis by bilateral partial nephrectomy. AB - Bilateral xanthogranulomatous pyelonephritis is a rare condition with 7 previously reported cases, all of which required dialysis and/or resulted in death of the patient. We report a case in which bilateral partial nephrectomy was performed successfully for bilateral xanthogranulomatous pyelonephritis. The etiology, symptomatology, radiographic findings and treatment of this rare inflammatory condition are discussed with a brief review of the previously reported cases. PMID- 8417186 TI - Transitional cell carcinoma of the ureteral stump 23 years after radical nephrectomy for adenocarcinoma. AB - We report a case of simultaneous invasive transitional cell carcinoma in a ureteral stump and superficial bladder tumor occurring 23 years after ipsilateral radical nephrectomy for adenocarcinoma of the kidney. We review the literature on similar cases and discuss potential etiologies of tumor formation in the ureteral stump. PMID- 8417187 TI - Pyelovenous lymphatic migration of transitional cell carcinoma following flexible ureterorenoscopy. AB - Ureterorenoscopy has enhanced the diagnosis and treatment of upper urinary tract disorders. However, the irrigation necessary for visualization and distention of the lumen may increase renal pelvic pressure, resulting in pyelovenous lymphatic backflow. We report a case of renal pelvic transitional cell carcinoma that may have migrated to submucosal venous and lymphatic spaces during flexible ureterorenoscopy. PMID- 8417188 TI - Nonoperative management of post-transplantation bladder leak: serendipitous salutary effect of temporary cyclosporine nephrotoxicity and oligoanuria. AB - Post-transplantation bladder leak, a potentially serious complication, is traditionally managed by reexploration and closure, and may require percutaneous placement of a nephrostomy tube. We report intractable bladder leakage that persisted following reclosure in a patient who also had cyclosporine nephrotoxicity. The attendant oligoanuria obviated the need for nephrostomy drainage and allowed healing of the bladder leak. The patient subsequently recovered from cyclosporine injury and regained renal function. PMID- 8417189 TI - Synchronous adenocarcinoma and transitional cell carcinoma of the bladder associated with augmentation: case report and review of the literature. AB - We report a case of synchronous transitional cell carcinoma and adenocarcinoma in an augmented bladder 8 years after cecocystoplasty. We discuss the clinical presentation as well as current clinical and basic research concepts, and suggest that the anastomotic zone in such cases may be at risk for malignant transformation. PMID- 8417190 TI - Delayed high flow priapism: pathophysiology and management. AB - Two cases of the management and pathophysiology of high flow arterial priapism are presented. Both cases were post-traumatic with delayed onset of priapism and both had angiographically diagnosed arteriocorporeal fistulas. Case 1 was managed with selective cavernous artery ligation and case 2 resolved spontaneously, both with excellent return of premorbid levels of erectile function. We propose that the pathophysiological mechanism involves injury to the intracavernous artery, causing ischemic necrosis. After a delay the arterial segment blows out, leading to unregulated blood flow into the corpus cavernosum. Management with surgical ligation is highly selective and nondisruptive to unaffected vessels, offering precise control of the bleeding vessel. PMID- 8417191 TI - Treatment philosophy and retreatment rates following piezoelectric lithotripsy. AB - Second generation lithotriptors offer the advantage of anesthesia-free fragmentation of renal and ureteral calculi but they frequently require multiple treatments to attain a stone-free status. However, excessive single lithotripsy sessions or multiple treatments may be associated with significant damage to the kidney. For some clinicians a common treatment philosophy involves evaluation of serial plain abdominal films every 24 hours after lithotripsy and immediate retreatment of all patients with incomplete fragmentation. To avoid unnecessary retreatments and, thus, minimize potential renal damage, we prospectively evaluated 100 patients undergoing lithotripsy on a Wolf Piezolith 2300 device. Patients were routinely treated with 4,000 shocks at 1,100 bar. Serial plain abdominal films were obtained at 1 day and 2 weeks after lithotripsy. The need for retreatment was determined by the plain abdominal film results. Additional therapy was considered necessary if there was no stone fragmentation or if residual fragments measured greater than 4 mm. Of the patients whose plain abdominal film at 24 hours indicated the need for a repeat treatment 43% were stone-free on the 2-week film. Thus, these patients were spared an unnecessary treatment by allowing adequate time for the stone fragments to pass spontaneously. Our data suggest that repeat treatments on second generation lithotriptors should not be performed within 24 hours. Rather, the patient should be reevaluated at least 1 to 2 weeks later to avoid unnecessary retreatment with the attendant potential for renal injury. In addition, when comparing the retreatment rates of various lithotriptors, one should also consider the treatment philosophy used at the particular institution and the timing of the radiographic studies used to determine the stone-free status. PMID- 8417192 TI - Osseous fixation of a penile prosthesis after transsexual phalloplasty: a case report. AB - We report on a patient who had undergone female-to-male transsexual surgery and subsequent phalloplasty by means of a free latissimus dorsi muscle graft with pudendal nerve coaptation elsewhere 10 years ago. The surgical strategy for implantation and osseous fixation of a penile prosthesis is described. We discuss whether the primary implantation of a penile prosthesis during 1-stage surgical phalloplasty is more advantageous in comparison with the currently favored secondary implantation. Phalloplasty should be represented by a 1-stage surgical procedure encompassing the creation of a neourethra, restoration of tactile and possibly erogenous sensibility, and implantation of a penile prosthesis. The result should have aesthetic value and be pleasing to the patient. PMID- 8417193 TI - Detection of the testis determining factor in an XX man. AB - An XX male patient was examined for the presence of 25 loci on the Y chromosome. Only 2 loci, the proximal border of the pseudoautosomal region Y and the sex determining region Y, were detected in this patient. The other 23 loci, including the zinc finger protein Y, were absent. We presume that a crossing over between the X and Y chromosomes occurred at the region proximal to the sex determining region Y but distal to the zinc finger protein Y during meiosis of the father. PMID- 8417194 TI - Mature testicular teratoma with vena caval invasion presenting as pulmonary embolism. AB - We report a case of mature testicular teratoma with invasion and thrombosis of the inferior vena cava that presented as recurrent pulmonary embolism. Treatment included radical orchiectomy, chemotherapeutic cytoreduction and, finally, resection of a massive retroperitoneal tumor with en bloc resection of the inferior vena cava using cardiopulmonary bypass and deep hypothermic circulatory arrest. Management is discussed and the literature is reviewed. PMID- 8417195 TI - Paratesticular myxoma: case report and review. PMID- 8417196 TI - Epididymal sarcoidosis. AB - Sarcoidosis is a multisystem disorder that rarely involves the genitourinary tract. To date only 28 cases of histologically proved sarcoidosis involving the epididymis have been described in the literature. Although uncommon, sarcoidosis should be considered in any differential diagnosis of testicular lesions. We present 2 cases of epididymal sarcoidosis. A 27-year-old asymptomatic black man had multiple nontender scrotal nodules on routine physical examination. A 34-year old black man was initially diagnosed with sarcoidosis by transbronchial biopsy. He received 10 months of prednisone therapy before noticing a mass in the right testicle. Surgical exploration of both patients demonstrated noncaseating granulomatous inflammation consistent with sarcoidosis. PMID- 8417197 TI - Squamous cell carcinoma of the prostate: 2 cases of a rare malignancy and review of the literature. AB - Primary squamous cell carcinoma of the prostate is an extremely uncommon malignancy, accounting for less than 1% of all prostatic cancers. We report on 2 patients with primary squamous cell carcinoma of the prostate: 1 with organ confined disease and 1 with metastatic disease. Both patients presented with urinary obstructive symptoms and carcinoma was not suspected on digital rectal examination. Serum acid phosphatase and prostate specific antigen levels were normal. From a review of the literature and our 2 cases it is apparent that squamous cell carcinoma of the prostate is biologically more aggressive than adenocarcinoma. PMID- 8417198 TI - Persistent vasocutaneous fistula associated with chronic urinary Mycobacterium chelonei infection. PMID- 8417199 TI - Postoperative clostridium difficile gastroenteritis. AB - Clostridium difficile gastroenteritis can be the cause of an enigmatic postoperative syndrome of high temperature and marked leukocytosis, out of proportion to the initially mild constitutional symptoms. Patients may suffer delayed onset of diarrhea, which will test positive for the C. difficile enterotoxin by latex agglutination. We report 5 cases of C. difficile gastroenteritis that occurred within a 2-year period. We believe that the combination of preoperative bowel preparation, and intraoperative and postoperative systemic antibiotics is the primary operant factor. All patients responded rapidly when oral antibiotics specific for C. difficile were instituted. The sequelae of C. difficile colitis can include toxic megacolon with perforation and peritonitis, increasing the importance of early recognition and appropriate treatment. PMID- 8417200 TI - Observations on anatomical aspects of the ureterovesical junction of the pig. AB - The paper reports the anatomy of the ureterovesical junction in pigs without urinary tract disease and the changes that occur with ageing. In comparison with other mammals the pig has a long intravesical ureter. Its length increases with age, from a mean length of 5 mm. at birth to 36 mm. at maturity. The width of the ureteric orifice also increases with age. The ureteric orifice was horseshoe shaped in 96.5% of cases, the remaining orifices were stadium shaped. The delineation of the anatomy of the porcine UVJ is important in the study of porcine and human pyelonephritis as the pig urinary tract is widely used as a human model. PMID- 8417201 TI - The effect of diabetes on sexual behavior and reproductive tract function in male rats. AB - The effect of streptozotocin induced diabetes and sabeluzole (SBZ) on sexual function was evaluated in male rats. SBZ is a benzothiazole derivative with antihypoxic and antiischaemic activities. Rats were rendered diabetic by intraperitoneal injection of streptozotocin, 60 mg./kg. body weight, and either left untreated or treated with 1.0 mg./kg. of SBZ. Two groups of control rats treated with or without SBZ were also evaluated. Seven weeks after the induction of diabetes, all rats were studied in vivo for mating behavior. Animals were sacrificed one week later, and detrusor strip response in vitro was evaluated. The reproductive organ weight, sperm content and motility as well as in vitro testosterone secretion and serum levels of LH and testosterone were determined. Diabetes induced significant reduction in mating behavior. The diabetic rats that received SBZ showed a significant improvement in mating behavior. The percentage of animals that exhibited ejaculation was 0% in the diabetic group compared to 70% in the controls and 38% in diabetic plus SBZ group. The strips of the detrusor muscle of the diabetic group showed a marked hypersensitivity to bethanechol HCL. In the diabetic plus SBZ group, the strips of the detrusor muscle showed a response similar to that of the control. The diabetic rats had significantly diminished reproductive organ weight, testicular sperm content, epididymal sperm content and sperm motility relative to the control. In addition, marked decrease in the serum level of testosterone and in vitro testosterone secretion was observed in diabetic rats. In the diabetic plus SBZ group, the reproductive organ weight, sperm content and motility as well as serum testosterone and in vitro testosterone secretion showed an improvement compared to diabetic rats. In summary, our data suggest that sex behavior and reproductive tract functions are markedly affected by streptozotocin induced diabetes. Sabeluzole treatment could be beneficial in reducing the deleterious effect of diabetes on sexual functions. PMID- 8417202 TI - The Swiss Lithoclast: a new device for endoscopic stone disintegration. AB - Even in the presence of extracorporeal shock wave lithotripsy there is still the need for endourological treatment of stones in the urinary tract. Stone fragmentation usually is achieved with either ultrasonic, electrohydraulic or laser lithotripsy. We report our early experience with a new, simply constructed machine at a reasonable cost for endoscopic stone disintegration--the Swiss Lithoclast. The principle of this lithotriptor is based on pneumatic shock waves induced by the central compressed air system of a hospital or by a compressor. This device was used to treat 151 patients with stones in the kidney, ureter, bladder or a Kock pouch continent urinary diversion. Endoscopic fragmentation was successful in all patients. Independent of the composition, all stones were disintegrated within a short period, indicating that the device may well represent an attractive alternative to standard endoscopic lithotriptors. PMID- 8417203 TI - Urodynamic effects of intravesical instillation of atropine and phentolamine in patients with detrusor hyperactivity. AB - The effects of intravesical instillation of atropine (10(-6) M) and phentolamine (10(-6) M) on urodynamic parameters were investigated in patients with detrusor hyperactivity of neurogenic or non-neurogenic origin. A modified cystometric technique with slow intermittent filling was used. The reproducibility of the investigative procedure was first verified in 17 patients. Eighteen patients, 12 with detrusor hyperreflexia and 6 with detrusor instability, were then investigated with atropine and 17 patients, 11 with detrusor hyperreflexia and 6 with detrusor instability, were investigated with phentolamine. In individual patients clinically relevant improvements, such as an increased bladder capacity and a decreased detrusor pressure, were found. Five/12 neurogenic and 1/6 non neurogenic patients responded to instillation of atropine. Five/11 neurogenic but none of the non-neurogenic patients responded to phentolamine. In the neurogenic group phentolamine caused an increase of the bladder capacity from 247 +/- 57 ml. to 378 +/- 57 ml. (p < 0.05). It is suggested that the different sensitivities to intravesical drug administration in patients with detrusor hyperreflexia and detrusor instability can be explained by pathophysiological differences and concluded that intravesical instillation of drugs may be of therapeutic value in selected groups of patients. PMID- 8417204 TI - Pseudomonas exotoxin conjugated to monoclonal antibody MRK16 specifically kills multidrug resistant cells in cultured renal carcinomas and in MDR-transgenic mice. AB - Using renal carcinoma and prostate carcinoma cell lines, we investigated the concept of targeting and killing multidrug resistant cells in urogenital cancers. Renal carcinoma lines HTB44, 45, 46, and 47 expressed a relatively low, but detectable level of multidrug resistance (MDR)1 mRNA as indicated by Northern blot analysis, whereas prostate lines LNCaP and DU145 were found to be MDR1 negative. Anti-P-glycoprotein monoclonal antibody MRK16 was conjugated to Pseudomonas exotoxin (PE) by a stable thioether bond. Treatment with MRK16-PE resulted in a dose-dependent killing of multidrug resistant renal carcinoma cells, while non-MDR expressing prostate carcinoma cells were not affected. Addition of excess MRK16 blocked the effect of MRK16-PE. Furthermore, MOPC-PE, a non-MDR associated monoclonal antibody control conjugate, did not target and kill multidrug resistant renal carcinoma cells. Having established that MRK16-PE was active against and specific for multidrug resistant cells in culture, we also tested bioactivity in MDR-transgenic mice, whose bone marrow cells express the human MDR1 gene at a level approximately equal to that found in many human cancers. Again, MRK16-PE killed multidrug resistant bone marrow cells with high efficiency in an intact animal, and killing was blocked by unconjugated MRK16. PMID- 8417205 TI - Antitumor effects of bacillus Calmette-Guerin in a syngeneic rat bladder tumor model system, RBT323. AB - We have studied the antitumor effects of Bacillus Calmette-Guerin (RIVM strain) in the syngeneic rat bladder tumor model RBT323. In an immunohistochemical infiltrate study we compared the antitumor effects of BCG with the immunopathological findings in order to get more insight into the possible effector mechanisms of BCG. The antitumor effects of BCG appeared not to be dose dependent, in the dose range tested. In rechallenge experiments no difference in growth of control tumors was seen between rats pretreated- or not pretreated with BCG. There was, however, a significant increase in antitumor effect of BCG after pretreatment with BCG. Immunohistological examination of BCG treated tumors revealed infiltrates consisting of macrophages, B-cells and T-cells. These results imply that whereas no specific response against the RBT323 cells is generated by BCG treatment, possibly BCG induced antigens do serve as immunogens in this nonimmunogenic syngeneic rat bladder tumor model system. PMID- 8417206 TI - Vasovasostomy in rabbits using fibrin adhesive prepared from a single human source. AB - We were interested in determining whether fibrin glue derived from a single human source could be used effectively to produce a sutureless vasovasostomy in the rabbit model. Fifteen rabbits were divided into two groups (7 control, 8 experimental) and underwent vasal transection and reanastomosis by conventional microsurgical suture techniques (control group) and sutureless anastomosis using single source fibrin glue (experimental group). The patency and tensile strength of the anastomoses were compared at four weeks and the anastomotic sites underwent histologic examination. This study demonstrated that a vasovasal anastomosis in the rabbit could be achieved using human single source fibrin glue with patency equivalent to a standard sutured anastomosis. The tensile strength of the glued anastomosis at four weeks was greater than the tensile strength of the sutured anastomosis. PMID- 8417207 TI - Renal vascular response to vasodilators following warm ischemia and cold storage preservation in dog kidneys. AB - The purpose of this study was to determine whether warm ischemia (WIT) and cold storage preservation (CSP) impair endothelium-dependent vascular relaxation in the kidney. Twenty-four canine kidneys were harvested, preserved with CSP for 24 or 48 hours, and then perfused with canine blood at 37 C for the determination of glomerular filtration rate (GFR), perfusion flow rate, and renal vascular resistance (RVR). There were four experimental groups: Group I--no WIT followed by 24 hours CSP, Group II--30 minutes WIT followed by 24 hours CSP, Group III--no WIT followed by 48 hours CSP, Group IV--30 minutes WIT followed by 48 hours CSP. Endothelial function in each group was evaluated using acetylcholine (ACh, 1 mg. bolus) as an endothelial dependent vasodilator, and sodium nitroprusside (NP, 10 mg. bolus) as an endothelial independent vasodilator. Glomerular filtration rate was significantly less (P < .05) and RVR was significantly greater (P < .05) for kidneys from Groups II, III and IV compared to group I. The highest RVR was observed in kidneys from Groups II and IV. Nitroprusside administration caused an equivalent reduction in RVR among all four study groups. ACh administration caused a similar reduction in RVR in Groups I and III; however, the change in RVR was significantly less in Groups II and IV (P < .05). We hypothesize that the more severe ischemic insult in the latter groups led to vascular endothelial damage with a consequent loss of ability to secrete endothelium-derived relaxing factor in response to ACh administration. PMID- 8417208 TI - Surgery following response to interferon-alpha-based therapy for residual renal cell carcinoma. AB - The role of aggressive surgery for metastatic renal cancer in the era of biological therapy is not clear. Therefore, we reviewed 17 patients who, between February 1987 and August 1990, underwent surgical resection of residual masses following initial response to interferon-alpha-based therapy. Viable tumor persisted in 15 patients (88%), whereas only inflammatory response was detected in 2. Of the patients 11 (65%) remained disease-free at a median of 12 months postoperatively (range 5 to 29), with an overall median survival of 26 months (range 6 to 34) from treatment initiation. These data suggest that surgery may be of therapeutic benefit in select patients with renal cell carcinoma who do not meet traditional criteria for surgical resection. Prospective trials are being performed to determine whether there is a role for aggressive surgical resection of persistent disease in patients with advanced renal cell carcinoma who have previously responded to interferon-alpha-based therapy. PMID- 8417209 TI - Bladder smooth muscle cells in culture: I. Identification and characterization. AB - This report documents the growth and culture characteristics of human and fetal bovine bladder smooth muscle cells in vitro. Bladder smooth muscle cell strains have been identified by their spindle shaped morphology, noncontact inhibited growth characteristics and the expression of smooth muscle cell specific alpha actin. Extracellular matrix protein biosynthesis by these cells in vitro has been characterized by metabolic labeling of proteins with [14C] radiolabeled proline and analysis by SDS gel electrophoresis. These studies demonstrate that bladder smooth muscle cells synthesize predominantly types I and III collagen, and fibronectin. In addition type III collagen exists in both a partially processed (pN alpha 1[III]) form and processed form. Complementary immunohistochemical studies show localization of type I, III, and IV collagens, and fibronectin to bladder smooth muscle cell extracellular matrix. We conclude that both fetal bovine and human smooth muscle bladder cells are capable of secreting the classic components of the surrounding connective tissue. PMID- 8417210 TI - A modified thoracoabdominal approach for retroperitoneal lymphadenectomy. AB - A modified thoracoabdominal approach for radical retroperitoneal lymphadenectomy is described. In addition, this same incision is ideal for surgical access to the retroperitoneal structures. The primary advantage of this approach is full exposure of the retroperitoneum with minimal retraction by assistants and without entering the chest cavity. PMID- 8417211 TI - Urinary dysfunction in Lyme disease. AB - Lyme disease, which is caused by the spirochete Borrelia burgdorferi, is associated with a variety of neurological sequelae. We describe 7 patients with neuro-borreliosis who also had lower urinary tract dysfunction. Urodynamic evaluation revealed detrusor hyperreflexia in 5 patients and detrusor areflexia in 2. Detrusor external sphincter dyssynergia was not noted on electromyography in any patient. We observed that the urinary tract may be involved in 2 respects in the course of Lyme disease: 1) voiding dysfunction may be part of neuro borreliosis and 2) the spirochete may directly invade the urinary tract. In 1 patient bladder infection by the Lyme spirochete was documented on biopsy. Neurological and urological symptoms in all patients were slow to resolve and convalescence was protracted. Relapses of active Lyme disease and residual neurological deficits were common. Urologists practicing in areas endemic for Lyme disease need to be aware of B. burgdorferi infection in the differential diagnosis of neurogenic bladder dysfunction. Conservative bladder management including clean intermittent catheterization guided by urodynamic evaluation is recommended. PMID- 8417212 TI - Urinary glycosaminoglycan excretion as a laboratory marker in the diagnosis of interstitial cystitis. AB - Urinary macromolecular uronate and glycosaminoglycan uronate concentrations were determined in 209 urine specimens obtained from 192 interstitial cystitis patients, 47 asymptomatic normal individuals and 32 spinal cord injury patients. As a group the concentration of macromolecular uronate or glycosaminoglycan uronate was significantly less in the interstitial cystitis patients than in the normal controls. Some interstitial cystitis patients showed low values of macromolecular uronate and glycosaminoglycan uronate. Analysis of the population distributions suggested that macromolecular uronate may have significant value in diagnosis of interstitial cystitis. PMID- 8417213 TI - Grading of superficial bladder cancer by quantitative mitotic frequency analysis. AB - A cohort of 270 superficial (stages Ta to T1) transitional cell bladder tumors was followed for more than 8 years. World Health Organization (WHO) grade, papillary status and 2 mitotic indexes were related to progression, recurrence free survival and bladder cancer related survival during followup. Mitotic activity index and volume corrected mitotic index were significantly related to WHO grade and papillary status (p < 0.0001). WHO grade, papillary status and mitotic indexes were related significantly to progression in univariate analysis (p < 0.001) whereas in a multivariate analysis only volume corrected mitotic index included independent prognostic information (p < 0.001). Recurrence-free survival was related to volume corrected mitotic index in the entire cohort (p = 0.03) and in papillary tumors (p = 0.07). Bladder cancer related survival was related to WHO grade, papillary status, mitotic activity index and volume corrected mitotic index (all p < 0.0001). In papillary tumors mitotic activity index (p < 0.0001), volume corrected mitotic index (p < 0.0001) and WHO grade (p = 0.0036) predicted survival. In multivariate analysis mitotic activity index predicted independently recurrence-free survival in the entire cohort (p = 0.043) and in papillary tumors (p = 0.012). Bladder cancer survival in the entire cohort and in papillary tumors was related independently to volume corrected mitotic index (p < 0.001). The results show that superficial transitional cell bladder tumors can be efficiently categorized into prognostic groups by quantitative mitotic frequency analysis and the results provide a new classification system for superficial transitional cell bladder tumors. PMID- 8417214 TI - Image cytometric deoxyribonucleic acid analysis of urine specimens as an adjunct to visual cytology in the detection of urothelial cell carcinoma. AB - We evaluated image cytometric deoxyribonucleic acid ploidy analysis of urine sediments as an adjunct to visual cytology in the detection of urothelial cell carcinoma. Both procedures were performed on 384 specimens obtained from voided urine (78%), bladder washings or by catheterization. Of these specimens 235 came from patients with active urothelial cell carcinoma, 23 from patients with currently inactive urothelial cell carcinoma and 126 from patients with benign disorders of the urinary tract. Visual cytology classified 171 specimens as positive for urothelial cell carcinoma, 143 as negative and 70 as atypical but diagnostically ambiguous. Sensitivity was 72.3% (170 of 235) and specificity was 99.3% (148 of 149, with atypical and negative specimens grouped together). The slides were then restained for ploidy analysis. Each of the deoxyribonucleic acid histograms of the 170 true positive and the 129 true negative specimens was described by a set of order statistics. These sets were entered into a multivariate discriminant analysis. The discriminant function obtained was then applied to the 70 ambiguous specimens. As a result 27 of these specimens, all from patients with active urothelial cell carcinoma, were reclassified as positive. Sensitivity was increased to 83.8% (197 of 235), with no loss of specificity. Image cytometry is a useful adjunct to visual cytology for the detection of bladder cancer in urine sediments. PMID- 8417215 TI - The AMS 700 inflatable penile prosthesis: long-term experience with the controlled expansion cylinders. AB - A total of 214 patients underwent implantation of the AMS 700 penile prosthesis with the controlled expansion cylinders. Followup ranged from 9 to 86 months, with a mean of 56 months. Life table analysis to calculate the probability of the cylinder and the device as a whole surviving 6 years revealed 97% for the cylinder and 90% for the device as a whole. No aneurysmal dilatation of the cylinders occurred. The incidence of cylinder leak was 0.7%. The infection rate in patients undergoing primary implantation of the device was 0.5% compared to 6.6% in patients undergoing revision surgery. The controlled expansion cylinders have significantly reduced the incidence of mechanical failures and, thus, the need for reoperation. As a consequence of the decreased need for reoperation the incidence of device infection has also decreased. PMID- 8417216 TI - Standardization of penile blood flow parameters in normal men using intracavernous prostaglandin E1 and visual sexual stimulation. AB - The evaluation of vasculogenic impotence by color flow Doppler ultrasound after injection of intracavernous vasoactive agents allows for simultaneous visualization in real time of arterial and venous blood flow. Normal arterial blood flow parameters after prostaglandin E1 injection have yet to be standardized. Our study was initiated to evaluate blood flow parameters in a normal control population after prostaglandin E1 and visual stimulation. A total of 20 healthy male volunteers 45 to 60 years old with histories of normal sexual function was selected. All volunteers were given intracavernous injections of 10 micrograms prostaglandin E1 and received concurrent visual stimulation by means of an erotic video. All patients developed rigid erections with no complications. Using color flow Doppler ultrasound measurements were done before and after prostaglandin E1 injection of right and left superficial and deep cavernous artery diameters, peak blood flow velocities and blood flow volumes. Results (mean plus or minus standard error) showed a significant increase in diameters after prostaglandin E1 in the superficial (20% increase) and deep (70% increase) penile arteries. Blood flow volume increased 3-fold for the superficial penile arteries (from 7.3 +/- 1.4 to 20 +/- 3.5 cc per minute) and 4-fold for the deep cavernous arteries (from 3.8 +/- 1 to 12.5 +/- 1.8 cc per minute). Peak blood flow velocity increased 2-fold (from 22 +/- 3 to 46 +/- 7 cm. per second) for the superficial arteries and 3-fold (from 12.5 +/- 2 to 37 +/- 5 cm. per second) for the deep cavernous arteries. These data suggest control values for normal erectile function in middle-aged men as a 70% increase in deep cavernous artery diameter, a systolic peak blood flow velocity greater than 30 cm. per second and more than 10 cc per minute of blood flow volume. With these standards the clinician may assess, design and follow treatment strategies for vasculogenic impotence. PMID- 8417217 TI - Collagenase versus placebo in the treatment of Peyronie's disease: a double-blind study. AB - We investigated 49 men with Peyronie's disease in a prospectively randomized placebo controlled double-blind study, comparing the effects on plaque size and penile deformity of intralesional purified clostridial collagenase and saline placebo. For the group as a whole, treatment out-performed placebo (p < 0.007). When patients were analyzed with respect to disease severity, those with lesser deformity responded more favorably to treatment. The absolute angular change in patients responding to treatment was small. No significant side effects were noted within a 3-month followup. PMID- 8417218 TI - Stimulated reactive oxygen species generation in the spermatozoa of infertile men. AB - To investigate the possible similarities between the reactive oxygen generating systems of the leukocyte and the spermatozoa, and to identify possible biochemical differences between fertile and infertile patients, the effect of various stimulants on the production of reactive oxygen in fertile, fertile with varicocele and infertile patients was examined. Generation of reactive oxygen species by human spermatozoa was examined in 2 patient groups: a fertile group (14), which included 7 patients with a palpable varicocele, and an infertile group (16) composed of 7 patients with a palpable varicocele and 9 who remained infertile 6 months to 3 years (mean 20 months) after internal spermatic vein ligation. Various known stimulants of reactive oxygen generation in leukocytes were used to assess spermatozoal reactive oxygen production. Reactive oxygen species generation was measured by the technique of chemiluminescence. In the infertile group reactive oxygen generation was markedly increased by the calcium ionophore A23187 and the chemoattractants N-formyl-methionyl-leucyl-phenylalanine and complement 5a (p < 0.05). In addition, fertile patients with varicoceles showed significantly enhanced chemoattractant stimulated reactive oxygen generation compared to fertile donors without varicoceles. This finding of a biochemical alteration in the sperm of infertile patients and patients with varicoceles may lend support to the argument for early varicocele repair. PMID- 8417219 TI - Impact of autologous blood transfusions on patients undergoing radical prostatectomy using hypotensive anesthesia. AB - We report on a retrospective study consisting of 71 consecutive patients who underwent radical retropublic prostatectomy under controlled hypotensive anesthesia, examining the impact of a preoperative autologous blood collection program on perioperative blood requirements. The population consisted of 34 patients who elected not to use autologous blood (group 1) and 37 patients who underwent radical prostatectomy after storing 1 to 3 units of their own blood (group 2). Median operative time and estimated blood loss were not statistically different. In group 1, 20 homologous blood units were used by 7 patients (21%). In group 2, 37 of 41 units transfused were autologous units and only 3 patients (8%) received homologous blood. However, for every unit of autologous blood deposited preoperatively there was a corresponding decrease in admission blood concentrations and 21 of 37 patients were anemic at hospitalization. In addition, the study suggests that in many patients the anemia produced by preoperative phlebotomy does not resolve preoperatively. In response, the effectiveness of preoperative autologous blood collection is decreased by this preoperative anemia. Although inefficient, we nevertheless conclude that an autologous blood collection program decreases homologous transfusion exposure and efforts should be directed to increase the erythropoietin response to the anemia produced by preoperative phlebotomy. Presently, it is an expensive program that offers only a modest benefit for patients undergoing radical retropubic prostatectomy. PMID- 8417220 TI - Cutaneous anesthesia with lidocaine-prilocaine cream: a useful adjunct during shock wave lithotripsy with analgesic sedation. AB - A comparative randomized double-blind study between 99 patients with (group 1) and 100 without (group 2) cutaneous anesthesia with a lidocaine-prilocaine cream during extracorporeal shock wave lithotripsy (ESWL*) with an unmodified Dornier HM3 lithotriptor was done. The application of the anesthetic cream or a placebo preparation was used in addition to premedication with meperidine hydrochloride and diazepam. In case of pain experience additional doses of analgesic sedation were administered. The requirement of additional analgesic sedation at a generator voltage of 14 kv. was significantly higher in group 2 than in group 1 (42 versus 26 patients, p < 0.05). The ESWL treatment was completed with only premedication in 39% of the patients in group 2 compared with 50% in group 1. The pain experience at 14 kv. was significantly lower for male patients in group 1 (p < 0.01). The most pronounced difference between groups 1 and 2 was recorded during treatment of stones in the upper calices, whereby 22% and 79%, respectively, required additional analgesic sedation at a generator voltage of 14 kv. (p < 0.05). A less pronounced but still significant difference was also recorded at 14 kv. for patients with stones in the other parts of the kidney and ureter (p < 0.05). The maximal median energy settings in groups 1 and 2 were 15.9 kv. and 15.4 kv., respectively. There were 27 patients in group 2 and 18 in group 1 who were treated with a maximum of 14 kv. The overall requirement of additional analgesic sedation was 62% in group 2 and 51% in group 1. Cutaneous anesthesia with lidocaine-prilocaine cream obviously has clinically significant effects on pain experience during ESWL. It cannot replace the need for analgesic sedation but it can be used advantageously to reduce the dosage of analgesic and sedative drugs during ESWL treatments performed without regional or general anesthesia in the unmodified Dornier HM3 lithotriptor. PMID- 8417221 TI - Why do sicker patients cost more? A charge-based analysis of patients undergoing prostatectomy. AB - Hospitals are reimbursed a greater amount for Medicare patients undergoing prostate surgery who have comorbid and complicating conditions than for patients without these conditions, since the former have been shown to have higher hospital costs and charges. We attempted to determine whether the higher hospital charges are due to duration of hospital stay and/or intensity of services. We analyzed hospital discharge data from 799 patients undergoing radical or transurethral prostatectomy during a 3-year period (1988 to 1991) at 2 major teaching hospitals by examining length of stay (duration), charges per hospital day (intensity) and total charges per stay. Mean lengths of stay were significantly longer for sicker versus healthier patients undergoing radical prostatectomy (7.4 versus 6.8 days at hospital 1 and 8.9 versus 7.8 days at hospital 2, p < 0.05) and transurethral prostatectomy (3.5 versus 2.8 days at hospital 1 and 3.5 versus 2.5 days at hospital 2, p < 0.05). Total hospital charges were significantly higher for sicker versus healthier patients undergoing radical prostatectomy ($14,557 versus $13,357 at hospital 1 and $17,864 versus $16,080 at hospital 2, p < 0.05) and transurethral prostatectomy ($6,446 versus $5,012 at hospital 1 and $5,468 versus $3,710 at hospital 2, p < 0.05). However, sicker and healthier patients had similar charges per day for radical prostatectomy ($1,959 versus $1,961 at hospital 1 and $2,006 versus $2,073 at hospital 2, p not significant) and for transurethral prostatectomy ($1.839 versus $1,800 at hospital 1 and $1.544 versus $1,488 at hospital 2, p not significant). On specified hospital days the charges per day for room/nursing, medical/surgical supplies, laboratory services and pharmacy services were similar for patients with and without comorbid conditions. Patients who are more ill at admission remain hospitalized longer after prostatectomy. However, they do not receive more intense care during their stays. For these procedures duration and not intensity appears to be the primary determinant of higher hospital charges for sicker patients. PMID- 8417222 TI - The use of mineral oil to manage the nondeflating Foley catheter. AB - The Foley catheter provides for urinary drainage by a self-retaining inflatable balloon mechanism. The morbidity associated with its use is minimal. However, blockage of the balloon port occasionally occurs, making its removal difficult. We discuss the various strategies used for catheter removal and recommend the use of mineral oil as the first line of intervention because of its reliability, safety and cost-effectiveness. PMID- 8417223 TI - Catheter balloon rupture using transrectal ultrasound. PMID- 8417224 TI - The use of iothalamate meglumine 17.2% as an effective testing medium in lower urinary tract urodynamic assessment of children. AB - Iothalamate meglumine 17.2% was compared to saline as a testing medium in 72 consecutive infants and children undergoing lower urinary tract urodynamic assessment. A total of 97 studies was performed of which 85 were complete. Multiple urodynamic parameters were used to compare test results. Agreement in test results was noted when assessing the change in detrusor pressure in 68 studies (77.6%), leak or peak bladder pressure in 79 (92.9%) and bladder contractions on filling in 81 (95.3%). The difference noted between the 2 testing mediums when comparing these 3 parameters was not statistically significant. The cystometric bladder capacity did vary between iothalamate meglumine 17.2% and saline but no particular trend was identified. The difference in cystometric capacity calculated as a percentage change from the saline baseline was not statistically significant (p = 0.55). Iothalamate meglumine 17.2% can be used as a reliable testing medium for pediatric cystometry. PMID- 8417225 TI - Long-term indwelling bilateral ureteral stents for bilateral hydronephrosis of unknown etiology. AB - We describe a rare case of long-term indwelling bilateral ureteral stents that were replaced yearly for 7 years because of azotemia associated with bilateral hydronephrosis of undetermined etiology on radiological examinations. During year 5 the patient had an uneventful gestation with delivery of a normal neonate by cesarean section. Hydronephrosis remained improved for 2 years after withdrawal of the stents. PMID- 8417226 TI - New reports make recommendations, ask for resources to stem TB epidemic. PMID- 8417227 TI - Resurgence of tuberculosis prompts US search for effective drugs, expanded research effort. PMID- 8417228 TI - Increasing use of chewing tobacco, especially among younger persons, alarms Surgeon General. PMID- 8417229 TI - 'Fundamentally sound' FDA advisory committee system expects to use most improvement ideas. PMID- 8417230 TI - From the Centers for Disease Control and Prevention. Tuberculosis transmission in a state correctional institution--California, 1990-1991. PMID- 8417231 TI - From the Centers for Disease Control and Prevention. Factors potentially associated with reductions in alcohol-related traffic fatalities--1990 and 1991. PMID- 8417232 TI - Malaria therapy: the value of the randomized controlled trial. PMID- 8417233 TI - The risk of sidestream smoke from pipes. PMID- 8417234 TI - Risk-related health insurance. PMID- 8417235 TI - Cumulative meta-analyses and the problem of multiple drug effects. PMID- 8417236 TI - Pseudohypertension and the Osler maneuver. PMID- 8417237 TI - What do women want? Comfy shoes--sometimes. PMID- 8417238 TI - Botulinum toxin: an alternative to handgun control? PMID- 8417239 TI - Decline of childhood Haemophilus influenzae type b (Hib) disease in the Hib vaccine era. AB - OBJECTIVE: Effective Haemophilus influenzae type b (Hib) conjugate vaccines were first licensed for use in US children at least 18 months old in December 1987 and for infants at least 2 months old in October 1990. We evaluated trends in Hib disease associated with licensure of Hib conjugate vaccines. DESIGN: Data from two sources, an intensive laboratory-based active surveillance system and the National Bacterial Meningitis Reporting System (NBMRS), were used separately to evaluate disease incidence. Data from vaccine manufacturers on Hib vaccine doses distributed in the United States were compared with trends in Hib disease incidence. RESULTS: The age-specific incidence of Hib disease among children less than 5 years old decreased by 71% from 37 per 100,000 persons in 1989 to 11 per 100,000 persons in 1991 (active surveillance data). Haemophilus influenzae meningitis incidence decreased by 82% between 1985 and 1991 (NBMRS data). Increases in doses of Hib vaccine distributed in the United States coincided with steep declines in Hib disease. Both surveillance systems showed decreased rates of Hib disease in infants less than 1 year old before vaccine was licensed for use in this age group. Haemophilus influenzae type b disease incidence in persons at least 12 years old and pneumococcal meningitis incidence in children less than 5 years old did not change substantially during the same period; therefore, decreased Hib disease in children less than 5 years old is not likely to be explained solely by changes in surveillance sensitivity or decreases in bacterial disease due to changes in medical practice. CONCLUSION: Our data suggest that conjugate vaccines have already had a marked impact on the incidence of Hib disease in the United States, preventing an estimated 10,000 to 16,000 cases of Hib disease in 1991. The decline of disease in infants less than 1 year old before licensure for this age group warrants further investigation. PMID- 8417240 TI - Decreases in invasive Haemophilus influenzae diseases in US Army children, 1984 through 1991. AB - OBJECTIVE: To document trends in the incidence of invasive Haemophilus influenzae diseases in young children of US Army active duty soldiers following the publication of H influenzae type b immunization recommendations for young children between 1985 and 1990. DESIGN: A population-based surveillance of hospital discharge diagnoses for H influenzae invasive diseases. SETTING: Military and civilian medical treatment facilities around the world. PATIENTS: There was a mean population of nearly 200,000 US Army health care beneficiaries younger than 5 years of age each year of the study. OUTCOME MEASURES: Annual total and age-specific incidences of the six most common H influenzae invasive diseases in this population (meningitis, epiglottitis, septicemia, pneumonia, cellulitis, and septic arthritis). RESULTS: The annual number of cases of H influenzae invasive diseases decreased from a high of 188 in 1986 to 43 in 1991. The incidence in the age group at highest risk for H influenzae disease, those 6 to 11 months of age, decreased from 355 per 100,000 children in 1986 to 116 per 100,000 in 1991 (P < .0001, chi 2 for trend). The incidence for H influenzae meningitis declined from 59 per 100,000 children in 1986 to 6 per 100,000 in 1991 (P < .0001, chi 2 for trend). CONCLUSIONS: The decrease in H influenzae invasive diseases closely follows the dates of H influenzae type b vaccine licensure for use in progressively younger age groups. Some age groups, however, experienced a decline in disease rates even before becoming eligible for vaccination. PMID- 8417241 TI - Smoking cessation and decreased risk of stroke in women. AB - OBJECTIVE: To prospectively examine the relationship of time since stopping smoking with risk of stroke in middle-aged women. DESIGN: An ongoing prospective cohort of women with 12 years' follow-up data (1976 to 1988), in which information on smoking habits was updated every 2 years by postal questionnaire. POPULATION STUDIED: A total of 117,006 female registered nurses aged 30 to 55 years in 1976 and free of coronary heart disease, stroke, and cancer at baseline. MAIN OUTCOME MEASURES: Incident strokes (fatal and nonfatal), further subdivided into ischemic stroke, subarachnoid hemorrhage, and cerebral hemorrhage. RESULTS: The age-adjusted relative risk of total stroke among current smokers compared with never smokers was 2.58 (95% confidence interval, 2.08 to 3.19). The corresponding relative risk among former smokers was 1.34 (95% confidence interval, 1.04 to 1.73). For total and ischemic stroke, the excess risks among former smokers largely disappeared from 2 to 4 years after cessation. The same patterns of decline were observed regardless of number of cigarettes smoked, the age at starting, or the presence of other risk factors for stroke. CONCLUSIONS: The risk of suffering among cigarette smokers declines soon after cessation and the benefits are independent of the age at starting and the number of cigarettes smoked per day. PMID- 8417242 TI - Neuropsychological sequelae of cardiac arrest. AB - OBJECTIVES: Prospective and community-based studies on the cognitive outcome of out-of-hospital cardiac arrest have not been published. We studied prospectively the neuropsychological sequelae of cardiac arrest and evaluated the effects of nimodipine on them. DESIGN: Placebo-controlled, randomized, double-blind trial of nimodipine compared with placebo in out-of-hospital ventricular fibrillation. SETTING: Urban area of 500,000 inhabitants served by the physician-manned Advanced Life Support Unit of Helsinki. PATIENTS: A total of 155 successfully resuscitated consecutive patients out of 677 resuscitation attempts during 2 1/2 years. Sixty-eight survivors were examined by a neuropsychologist and a neurologist. MAIN OUTCOME MEASURE: Neuropsychological outcome 3 months and 1 year after cardiac arrest. INTERVENTIONS: Nimodipine or placebo at a dosage of 10 micrograms/kg as an intravenous injection immediately after restoration of spontaneous circulation, followed by an infusion of 0.5 micrograms/kg per minute for 24 hours. RESULTS: Three months after cardiac arrest, 41 (60%) of 68 patients were found to have moderate to severe cognitive deficits. At 12 months, 26 (48%) of 54 survivors still had moderate to severe deficits, and the Symptom Check List 90--Revised score indicated the presence of depression in 22 patients (45%) and severe depression in 12 patients (24%). CONCLUSIONS: Moderate to severe neuropsychological sequelae of out-of-hospital cardiac arrest are still present in approximately one half of the survivors at 1 year and may be permanent. There seems to be no excess of increased disability in the subgroup of patients with delayed advanced life support. Nimodipine failed to show any effect on the cognitive functions tested. PMID- 8417243 TI - Breast-feeding. Time to teach what we preach. PMID- 8417244 TI - Declining incidence of Haemophilus influenzae type b disease since introduction of vaccination. AB - OBJECTIVE: To determine the incidence of invasive Haemophilus influenzae type b disease in children living in Minnesota and Dallas County, Texas, before and since introduction of plain polysaccharide vaccine in 1985, and conjugate vaccine in 1988. Initially, use of these vaccines was limited to infants 18 months of age and older. DESIGN: Identification of culture-proven cases of H influenzae type b disease was through systems of active, laboratory-based surveillance. The mean incidence of disease (cases observed/100,000 child-years) for 1983 and 1984 served as a baseline for comparison with subsequent years through 1991. PARTICIPANTS: Children less than 5 years of age in Minnesota and Dallas County, Texas. Overall, there were 2557 confirmed age-eligible cases of invasive H influenzae type b disease from 1983 through 1991. RESULTS: Between the 1983-1984 baseline and 1991, the incidence of H influenzae type b disease decreased 85% in Minnesota and 92% in Dallas. Notably, declines in incidence were observed in children in the age group being vaccinated as well as in infants younger than 18 months of age prior to introduction of vaccination. CONCLUSION: In two widely separated areas of the United States, a profound decrease in the incidence of H influenzae type b disease has occurred. The data suggest that vaccination may be protecting against disease, as well as decreasing the spread of infection to unvaccinated infants. PMID- 8417245 TI - Cost-effectiveness of monoclonal antibodies to gram-negative endotoxin in the treatment of gram-negative sepsis in ICU patients. AB - OBJECTIVE: To evaluate the fiscal impact and the cost-effectiveness of monoclonal antibodies against gram-negative endotoxin (MAbGNE) in the treatment of presumed gram-negative sepsis. DESIGN: A decision analysis model was developed from (1) data from two phase III trials that studied the E5 or HA-1A MAbGNE, and (2) financial data from 1405 septic patients who required intensive care at a large tertiary hospital. SETTING: Intensive care unit (ICU) patients with presumed gram negative sepsis. PATIENTS: The E5 trial evaluated 468 patients, and the HA-1A study enrolled 543 patients with presumed gram-negative sepsis. INTERVENTIONS: The addition of MAbGNE to standard regimens or standard regimens alone. MAIN OUTCOME MEASURES: Total expected charges and the expected probability of survival were determined for each option. Cost-effectiveness and marginal cost effectiveness ratios were also derived. Multiple sensitivity and Monte Carlo analyses were performed to test the underlying assumptions. RESULTS: MAbGNE therapy always resulted in higher expected charges; however, these differences were less than its acquisition cost by $870. The cost-effectiveness ratio for MAbGNE, for $2000 and $4000 acquisition costs, was $71,674 and $74,900 per probability of survival, respectively. Sensitivity analysis showed that cost effectiveness was most affected by diagnostic accuracy, patient selection, and acquisition cost. Monte Carlo analysis showed that MAbGNE was more costly for 71% of simulations, yet the most efficacious option for 79% of simulations. CONCLUSIONS: From the perspective of acute care institutions, MAbGNE is expensive and cannot be justified on a cost-saving basis. However, it may be cost-effective throughout a reasonable range of assumptions. PMID- 8417246 TI - Controlling the resurgent tuberculosis epidemic. A 50-state survey of TB statutes and proposals for reform. PMID- 8417247 TI - Infections caused by Haemophilus influenzae type b. The beginning of the end? PMID- 8417248 TI - Perioperative evaluation of a patient with abdominal aortic aneurysm. PMID- 8417249 TI - Trends in the utilization of primary total hip arthroplasty, 1969 through 1990: a population-based study in Olmsted County, Minnesota. AB - Using the unique data resources of the Rochester Epidemiology Project, we identified the residents of Olmsted County, Minnesota, who underwent total hip arthroplasty (THA) between 1969, when this technique was introduced, and 1990. During this period, 735 residents underwent 859 primary THAs. An initial dramatic upsurge in utilization of THA soon after its introduction was followed by a lower rate of increase, to an age- and sex-adjusted rate of 59.9 per 100,000 person years in 1987 through 1990. In Olmsted County, the procedure was used more often in women than in men, but men underwent THA at a younger age than did women. THAs were performed for various conditions of the hip, but the increased utilization during the last decade of the study was mainly attributable to its application in primary degenerative hip disease among patients (male more often than female) from a wider age range; utilization of THA for hip fractures decreased during the study period. Most THAs were of the cemented variety (86.1%); however, the rate of use of noncemented THAs, first performed in 1984, is increasing. The overall upward trend in Olmsted County was consistent with European data but less than the rate of increase observed for the United States as a whole between 1983 and 1987. Olmsted County utilization rates, if projected nationally, suggest a need for more than 139,000 primary THAs each year at an annual direct cost of more than $2.7 billion. An urgent need exists for detailed examination of national utilization patterns and clarification of the role of cementless implants and of THA for hip fractures. PMID- 8417250 TI - Cardiovascular responses to immersion in a hot tub in comparison with exercise in male subjects with coronary artery disease. AB - In order to test the safety of hot tub use for persons with heart disease, 15 men with clinically stable coronary artery disease underwent 15 minutes of immersion in a hot tube at 40 degrees C. On another day, they exercised on a cycle ergometer for 15 minutes; target heart rate was determined by standard methods. Tympanic temperature, skin temperature, electrocardiographic findings, blood pressure, plasma catecholamines, subjective comfort, and cardiovascular symptoms were monitored. The peak heart rate was significantly lower during the hot tub session versus the exercise session (85 +/- 14 versus 112 +/- 19 beats/min), as were the systolic (106 +/- 15 versus 170 +/- 21 mm Hg) and diastolic (61 +/- 6 versus 83 +/- 8 mm Hg) blood pressure measurements (P < 0.01). Tympanic temperature increased by a mean of 0.6 +/- 0.3 degrees C during immersion and 0.1 +/- 0.1 degrees C during exercise. No ischemic electrocardiographic changes or clinical complications occurred. Simple ventricular ectopic activity and "just noticeable" chest pain were more frequent during exercise than during immersion. Plasma norepinephrine increased during exercise but not during immersion. These data suggest that hot tub use within these time and temperature constraints should be safe for men with stable heart disease who can follow an exercise regimen at home. PMID- 8417251 TI - "Benign" monoclonal gammopathy--after 20 to 35 years of follow-up. AB - All 241 patients with an apparently benign monoclonal gammopathy who were examined at the Mayo Clinic before Jan. 1, 1971, underwent prospective follow-up for 20 to 35 years (median, 22 years). Electrophoresis and immunoelectrophoresis of serum and urine specimens were performed periodically in an effort to determine the frequency of development of multiple myeloma, primary amyloidosis, macroglobulinemia, or other lymphoproliferative diseases. At follow-up, the patients were categorized into one of four groups: group 1 (benign)--46 patients (19%) who were alive and had a benign monoclonal gammopathy; group 2--23 patients (10%) who had a serum monoclonal protein value of 3 g/dl or more but did not require chemotherapy; group 3--113 patients (47%) who died without evidence of myeloma or related disorders; and group 4-59 patients (24%) in whom multiple myeloma (39), systemic amyloidosis (8), macroglobulinemia (7), or a malignant lymphoproliferative disease (5) developed at a median of 10, 9, 8, and 10 1/2 years, respectively, after detection of the monoclonal protein. Thus, in patients with an apparently benign monoclonal gammopathy, follow-up must be continued indefinitely because multiple myeloma, amyloidosis, macroglobulinemia, or related disorders occur in approximately a fourth of them. PMID- 8417252 TI - Relationship of hemoglobin level and duration of hospitalization after total hip arthroplasty: implications for the transfusion target. AB - We analyzed the relationship of hemoglobin level and duration of hospitalization of patients who underwent primary cemented total hip arthroplasty for degenerative joint disease at our institution. Retrospectively, we reviewed the medical records of 332 patients treated during a 16-month period (May 6, 1989, to Aug. 20, 1990). The following variables were analyzed: number of postoperative days to dismissal from the hospital, level of hemoglobin preoperatively and at dismissal, decrease in hemoglobin level from preoperatively to time of dismissal, patient age, surgeon, and blood products transfused. No correlation was found between level of hemoglobin at dismissal, preoperative hemoglobin level, or decrease in hemoglobin concentration from preoperatively to time of dismissal and number of days to dismissal from the hospital. Advanced age was associated with a longer hospital stay. A slight but statistically significant difference was noted in duration of hospitalization among patients operated on by different surgeons. Patients who received both autologous and homologous blood required more transfusion (3.8 units) and had a longer hospital stay (10.7 days) than did patients who received autologous blood only (2.4 units and 9.5 days) or homologous blood only (2.6 units and 10.2 days). We conclude that variation in hemoglobin levels among patients in our study was unrelated to duration of hospitalization. This finding suggests that transfusion of autologous or homologous blood to achieve a higher hemoglobin level (higher transfusion target) solely for shortening hospital stay is unwarranted. PMID- 8417253 TI - 100th birthday of Dr. Samuel F. Haines. PMID- 8417254 TI - Increased plasma level of endothelin-1 in the Okamoto spontaneously hypertensive rat. AB - The objectives of this study were to determine plasma levels of endothelin (ET) in a genetic model of hypertension and in control rats during control conditions and in response to short-term volume expansion with saline. Okamoto spontaneously hypertensive rats (SHR) and control Wistar-Kyoto (WKY) rats were used in this study. One group of each strain served as control animals, and another group of each strain underwent volume expansion with saline (5% of body weight infused during a period of 30 minutes). The levels of ET-1 and ET-3 were measured in plasma by using a double-antibody radioimmunoassay. In the control groups of SHR and WKY rats, plasma ET-1 levels were 72.5 +/- 14.9 pg/ml (N = 8) and 40.2 +/- 7.5 pg/ml (N = 12), respectively (P < 0.05). In the volume-expanded SHR group (N = 8), the plasma ET-1 level was 45.5 +/- 11.1 pg/ml (approximately 37% less than that of the control SHR group), whereas it was 40.6 +/- 10.2 pg/ml in the volume expanded group of WKY rats (N = 10) (almost identical to that of the control WKY group). Plasma levels of ET-3 were similar in control and in volume-expanded groups of SHR and WKY rats. These data show that basal levels of plasma ET-1 are significantly higher in the SHR than in the WKY rat.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417255 TI - Chronic total obstruction and short-term outcome: the Excimer Laser Coronary Angioplasty Registry experience. AB - Percutaneous transluminal coronary angioplasty for chronic total obstructions is associated with significantly decreased success rates in comparison with those for dilation of subtotal stenoses. Failure usually results from inability to cross the occlusive lesion with a guidewire, although it may result from inability to pass the balloon catheter after the guidewire has been passed. In the Excimer Laser Coronary Angioplasty Registry, 172 chronic total obstructions were treated in 162 patients (10.3% of the 1,569 patients entered). For chronic total obstructions, passage of a guidewire is a prerequisite for laser angioplasty. Once a guidewire crossed an occlusion, the overall laser success rate for treatment of chronic total obstructions was 83%; the extent of stenosis decreased from 100% to 55 +/- 26%. Success was independent of length of the occlusive lesion. In 74% of patients, adjunctive percutaneous transluminal coronary angioplasty was used after laser angioplasty. A final procedural success, defined as residual stenosis of less than 50% and no major complication (coronary artery bypass grafting, myocardial infarction, or death), was achieved in 90%. Major complications were infrequent; 1.2% of patients required coronary artery bypass grafting, and 1.9% had a Q-wave myocardial infarction. Only one death occurred. The use of laser angioplasty may be of particular value when chronic total obstructions can be crossed with a guidewire but not with a conventional balloon catheter or when the occlusion is confirmed to be extremely long. PMID- 8417256 TI - Coronary restenosis: prospects for solution and new perspectives from a porcine model. AB - Coronary restenosis, a major unresolved problem for percutaneous coronary revascularization procedures, has thus far been resistant to all therapeutic strategies. In part, ineffective treatment or prevention of coronary restenosis may be due to reliance on a conceptualization of the restenosis process that incompletely reflects the pathophysiologic factors associated with neointimal formation after arterial injury. In a porcine coronary restenosis model, three stages of neointimal growth after arterial injury have been identified: an early thrombotic stage, with platelets, fibrin, and erythrocytes; a cellular recruitment stage, with endothelialization and an infiltration by lymphocytes and monocytes; and a proliferative stage, in which smooth muscle cells migrate into and proliferate within the fibrin-rich degenerating thrombus. Evaluation of basic mechanisms responsible for neointimal formation has been possible with this model. In particular, a direct relationship exists between the depth of arterial injury and subsequent neointimal thickness. This relationship can be used for investigating the efficacy of new therapies. Treatment strategies for restenosis should be directed toward interference with the cellular or humoral events that lead to neointimal formation, with the specific goal of decreasing the neointimal volume. These strategies may include delivery of drugs to the site of arterial injury to limit the amount of early mural thrombus or decreasing subsequent cellular recruitment and proliferation as well as synthesis of extracellular matrix. PMID- 8417257 TI - Resting thallium-201 scintigraphy for identifying viable myocardium in a patient with severe left ventricular dysfunction. AB - Recognizing patients with coronary artery disease in whom severe left ventricular dysfunction is attributed to a chronic decrease in myocardial blood flow without infarction is often difficult but important because such patients may benefit from surgical revascularization. Herein we describe a patient with severe left ventricular dysfunction who had appreciable resting wall motion abnormalities; tomographic thallium-201 myocardial scintigraphy performed while the patient was resting identified viable myocardium. Subsequent revascularization improved regional and global left ventricular function considerably. PMID- 8417258 TI - Harvey Williams Cushing--pioneer neurosurgeon. PMID- 8417259 TI - Hepatic allograft rejection: new developments in terminology, diagnosis, prevention, and treatment. AB - Hepatic allograft rejection remains a major cause of morbidity related to the need for increased immunosuppression and continues to be a principal cause of late failure of the graft. Hepatic allograft rejection is defined on the basis of morphologic findings; cellular rejection is defined as portal or periportal hepatitis with nonsuppurative cholangitis or endotheliitis (or both), and ductopenic rejection is defined as loss of interlobular and septal bile ducts, typically in at least 50% of the portal tracts. The overall incidence of episodes of cellular rejection, which usually occur within the first 2 weeks after liver transplantation, varies from 50 to 100%. Ductopenic rejection occurs in approximately 8% of patients who undergo initial liver transplantation and is usually diagnosed between 6 weeks and 6 months after transplantation. Induction and maintenance immunosuppression with triple-drug (cyclosporine, prednisone, and azathioprine) therapy and other combinations that include antilymphocyte preparations, however, has decreased the incidence of both cellular and ductopenic rejection. In patients experiencing episodes of cellular rejection, high-dose intravenously administered corticosteroid therapy yields the best response and is associated with a lower incidence of ductopenic rejection than is low-dose and orally administered corticosteroid therapy. The correlation between degree of biochemical liver dysfunction and presence of histologic rejection is minimal early after transplantation. Histologic severity of rejection, however, suggests which patients will require more immunosuppression and which patients may need antilymphocyte therapy for controlling the rejection episode. With the availability of new immunosuppressive agents, distinguishing patients at high risk for rejection is important. The goals for use of new immunosuppressive agents and regimens are to improve graft and patient survival, to decrease the incidence of cellular and ductopenic rejection, and to minimize side effects and complications. PMID- 8417260 TI - Apparent skepticism: capital punishment and medical evidence. AB - In recent cases on the constitutionality of sentencing to death criminals who were younger than 18 years of age at the time of their crimes or who are mentally retarded, the US Supreme Court has rejected medical evidence that such persons categorically possess diminished culpability. Rather, the Court has accepted the public's "apparent skepticism" of such a scientific consensus in upholding the execution of capital offenders who are 16 years of age or older. The 1952 English case of Craig and Bentley sparked discussion of similar issues in the United Kingdom and contributed to the abolition of capital punishment for murder in that country. US courts should have more deference for such medical evidence, despite perceived widespread resistance to the conclusions of researchers that adolescents and mentally retarded persons categorically lack sufficient maturity, judgment, and deliberation to receive capital punishment and that they are not deterred from murder by the threat of execution. PMID- 8417261 TI - Searching for therapeutic niches for new coronary interventional devices: the unyielding nature of chronic occlusions. PMID- 8417262 TI - What is the right rate for total hip arthroplasty? PMID- 8417263 TI - The restenosis "antitheory". PMID- 8417264 TI - Meta-analysis: its role in medical research and in assessment of the association between low levels of cholesterol and excess mortality. PMID- 8417265 TI - Silicone gel breast implants. PMID- 8417266 TI - Graphic interpretation of serum parathyroid hormone, calcium, and phosphorus values in primary hyperparathyroidism. PMID- 8417267 TI - Asset spend-down in nursing homes. Methods and insights. AB - The issue of how many elderly are affected by catastrophic nursing home expenses is a major part of the debate over if and/or how to reform long-term-care financing. Currently, there is some discussion regarding the magnitude of this catastrophic event, referred to as "asset spend-down", among the elderly. National data suggest the magnitude is small, while state-specific studies indicate it is greater. In addition, the literature regarding asset spend-down has presented two different measures of its magnitude, further confusing the issue. These two measures, each based on different denominators, have often been presented without adequate explanation. In this study, the authors review both measures and analyze reasons for the differences observed across studies. Major reasons identified include the type of sample used, the mix of payor source at admission, the length of time covered by the data, data on payor source/Medicaid eligibility, and the ability to observe multiple nursing-home stays within the data. Using the measure based on the number of persons who are private pay at admission, these studies indicate that approximately one fourth will eventually deplete assets. The second measure, based on a count of Medicaid residents at a point in time, indicates approximately one third were private pay when admitted. Study results indicate that national studies have underestimated the extent of spend-down due to national-level data limitations, while state-specific studies inevitably refect the specific state data set available and circumstances particular to each state. More state studies and a better understanding of asset transfer are needed. PMID- 8417268 TI - Self-care responses to symptoms by older people. A health diary study of illness behavior. AB - Although studies of illness behavior have concentrated on utilization of formal services, recent research has highlighted the importance of lay care. In this article, patterns of self-treatment of symptoms among a sample of older people living in community settings are explored. Data were collected through personal interviews and health diaries completed by a probability sample of 667 elderly people. A majority of respondents managed most symptoms on their own. A decision to take no action was a common response to a number of symptoms. Prescription and over-the-counter medications were the most frequently reported intervention strategies. Multivariate analyses affirmed the importance of both symptom interpretation and symptom experience on lay treatment decisions. Causal attributions and prior experience with symptoms exhibited a less consistent impact on lay care strategies. Whether people ignore or treat symptoms appears to have less to do with their familiarity and causal explanation than with whether or not the symptom causes them pain or discomfort, whether it interferes with their desired activities, or whether they think it might be indicative of a serious decision. PMID- 8417269 TI - Health plan switching in anticipation of increased medical care utilization. AB - We compare rates and days of maternity and nonmaternity hospital admission for the years 1981 through 1984 for three groups of employees and dependents from a large private employer: those continuously enrolled in a fee-for-service (FFS) plan (N = 147,700), those continuously enrolled in a health maintenance organization (HMO) (N = 30,957), and those switching from the FFS plan to the HMO (N = 2,144). The rate of maternity admissions for plan switchers increased by 106% (P < 0.001) in the post-switch year compared with the pre-switch year, while maternity rates for continuing FFS-plan enrollees declined by 12% (P < 0.001) and rates for continuing HMO enrollees remained unchanged. Nonmaternity admission rates for switchers decreased by 19% (P = 0.079), consistent with the expectation that HMOs reduce these rates substantially, while rates for FFS-plan stayers increased 4% (P < 0.001) and those for HMO stayers remained unchanged. We conclude that employees often switch health plans when anticipating increased needs for maternity care and therefore that pre-switch rates of utilization are unreliable measures of the true magnitude of risk selection between HMOs and FFS plans. PMID- 8417270 TI - Segmentation in local hospital markets. AB - This study examines evidence of market segmentation on the basis of patients' insurance status, demographic characteristics, and medical condition in selected local markets in California in the years 1983 and 1989. Substantial differences exist in the probability patients may be admitted to particular hospitals based on insurance coverage, particularly Medicaid, and race. Segmentation based on insurance and race is related to hospital characteristics, but not the characteristics of the hospital's community. Medicaid patients are more likely to go to hospitals with lower costs and fewer service offerings. Privately insured patients go to hospitals offering more services, although cost concerns are increasing. Hispanic patients also go to low-cost hospitals, ceteris paribus. Results indicate little evidence of segmentation based on medical condition in either 1983 or 1989, suggesting that "centers of excellence" have yet to play an important role in patient choice of hospital. The authors found that distance matters, and that patients prefer nearby hospitals, moreso for some medical conditions than others, in ways consistent with economic theories of consumer choice. PMID- 8417271 TI - A smoking-cessation intervention for hospital patients. AB - Many patients attempt to stop smoking during hospitalization, but most relapse after discharge. This study developed and evaluated a brief smoking-cessation and relapse-prevention program for hospitalized smokers. All hospitalized smokers (n = 1,119) were identified by questionnaire at hospital admission and then received either usual care or usual care plus a hospital-based smoking-cessation intervention regardless of interest in stopping smoking. Intervention components included a 20-minute bedside counseling session, a 12-minute videotape, a variety of self-help materials, and a follow-up telephone call. Special attention was given to techniques for preventing relapse after hospital discharge. Defining ex smokers as those who reported no tobacco use at both 3- and 12-month follow-up assessments, and counting those lost to follow-up as smokers, the intervention increased the proportion of patients who quit smoking by one half (9.2% vs 13.5%, P < 0.05). These results demonstrate the efficacy of a brief in-hospital intervention and suggest that relapse-prevention efforts are needed to convert temporary cessation during hospitalization into long-term abstinence. PMID- 8417272 TI - Specialty differences in the 'July Phenomenon' for Twin Cities teaching hospitals. AB - This study evaluated changes over the academic year in the cost and the outcome of inpatient care to investigate the effect of housestaff experience in teaching hospitals. Patients with 25 preselected discharge diagnoses, admitted between January 1, 1983 and December 31, 1987 to acute-care, nonfederal, non-pediatric hospitals in the Minneapolis/St. Paul metropolitan area (total number available for analysis 240,467) were examined. Level of housestaff experience was measured as the number of days (1 to 365) into the academic year when the patient was admitted. Linear and logistic regression analyses were used to evaluate the different effects of experience on patient care in teaching hospitals compared with nonteaching hospitals. For the subset of patients with internal medicine diagnoses, the expected "July Phenomenon" was observed, with significant relative declines in diagnostic and pharmaceutical charges in teaching hospitals over the academic year. In contrast, surgery patients showed an increase in length of stay and various charges over the academic year in teaching hospitals. There were no meaningful effects of housestaff experience on mortality, operative complications, or nursing home discharge. These results indicate that housestaff training is significantly related to the use of hospital resources for inpatients, but that the degree and direction of the effects differ by specialty. These findings may reflect important differences among training programs in the process of physician education and its effects on patient care. PMID- 8417273 TI - A causal model of health status and satisfaction with medical care. AB - Patients with better health status have often been shown to be more satisfied with their medical care, but the causal factors in this relation have not been determined. In this study, a longitudinal assessment of these two constructs was undertaken in which older patients in a health maintenance organization were interviewed at baseline (N = 590) and again 1 year later (N = 526) about their health status and satisfaction with their medical care. Structural equation modeling using LISREL procedures revealed that the predominant direction of causation went from earlier self-perceived overall health and functional ability to later levels of satisfaction. There was no evidence for causal paths going from satisfaction to later health. In addition, a test of spuriousness indicated that for self-perceived overall health, the significant longitudinal path was unlikely to be explained by unmeasured confounding variables. PMID- 8417274 TI - Effects of ethanol on punished and nonpunished responding under conditions of equated reinforcement rates and similar response rates. AB - Lever pressing by pairs of rats was maintained under random-ratio (first subject) and yoked-interval (second subject) schedules of food presentation. The inter reinforcement intervals generated under the ratio schedule comprised the interval values for the second (yoked) subject. This arrangement yielded nearly equivalent rates of food presentation for each subject pair. For the first rat of each pair a random-ratio schedule of shock presentation was added to the ratio schedule of food presentation. This manipulation resulted in similar rates of punished (first rat) and nonpunished (second rat) responding within subject pairs. Ethanol administration (0.25-1.5 g/kg) generally resulted in dose-related decreases in both punished and nonpunished responding. In general, punishment-specific effects were not obtained. These results suggest that ethanol may not be as effective as chlordiazepoxide or pentobarbital in increasing punished responding even when the effects of baseline response and reinforcement rates are controlled. PMID- 8417275 TI - Cytidylate cyclase activity is stimulated via activation of a guanine nucleotide binding protein. AB - Cytidylate cyclase is a membrane-bound enzyme which catalyzes the biosynthesis of cytidine 3',5'-cyclic monophosphate (cCMP) from CTP. By using a sensitive and specific enzyme immunoassay method, we evaluated the participation of guanine nucleotide-binding protein (G-protein) in the regulation of cytidylate cyclase activity in rat brain and other tissues. AlF4-, an activator of G-proteins, effectively elevated the cyclase activity. The stimulation by GTP gamma S, a nonhydrolyzable GTP analogue, was also observed in time- and concentration dependent manner during the preincubation and this effect was competitively inhibited by the addition of GDP beta S. However, islet-activating protein and cholera toxin which affected adenylate cyclase activity had no effect on cytidylate cyclase activity. These results indicate that cytidylate cyclase is associated with a certain G-protein and its activity is regulated by the mode different from that of adenylate cyclase. PMID- 8417276 TI - Determination of tolbutamide hydroxylation in rat liver microsomes by high performance liquid chromatography: effect of psychoactive drugs on in vitro activity. AB - A simplified HPLC method for tolbutamide metabolism to hydroxytolbutamide has been used to screen sixty psychoactive drugs for their ability to inhibit rat liver microsomal tolbutamide hydroxylation. One-step extraction with diethyl ether was followed by reconstitution and isocratic HPLC analysis with a binary mobile phase (ammonium phosphate:methanol, 45:55, v/v). Nanogram amounts of hydroxytolbutamide formation were estimated with UV detection at 240 nm. Hydroxytolbutamide formation was linear with incubation times of 40-120 min, but specific activity increased with increases in microsomal protein (0.15-1.10 mg). A differential inhibitory response was demonstrated for tolbutamide and debrisoquine hydroxylation to 5 psychoactive drugs, suggesting that tolbutamide hydroxylation is not dependent on P4502D1. Sixty psychoactive drugs, or drug metabolites, (at 33 microM) were then co-incubated with tolbutamide (at 2.5 and 10.2 microM). Tolbutamide hydroxylation was refractory (< 25% inhibition) to twenty-four of the drugs and only mildly inhibited (25-50% inhibition) by twenty eight. Two compounds, trans-3-methylfentanyl and flurazepam, produced > 50% inhibition that was independent of tolbutamide concentration. Five of the drugs (methadone, chlorpheniramine, meperidine, 6-monoacetylmorphine and methylphenidate), however, caused greater than 50% inhibition in a competitive manner which suggests these drugs may share an affinity for the substrate binding site for tolbutamide. PMID- 8417277 TI - Cytochrome P450TB (CYP2C): a major monooxygenase catalyzing diclofenac 4' hydroxylation in human liver. AB - The nature of the enzyme(s) catalyzing the major metabolic pathway of diclofenac, 4'-hydroxylation, was investigated in human liver microsomes. Inhibition studies were performed with tolbutamide and sulfaphenazole (respectively the prototype substrate and a selective inhibitor of cytochrome P450TB--CYP2C subfamily), and with phenytoin and (+/-)-warfarin, other proposed substrates of P450TB. Diclofenac 4'-hydroxylation displayed single enzyme Michaelis-Menten kinetics and was similar in microsomes from one poor and five extensive metabolizers of debrisoquin (CYP2D6), with a Km of 5.6 +/- 1.5 microM (mean +/- sd) and a Vmax of 60.6 +/- 23.5 nmol/mgP/h. Inhibition by tolbutamide, sulfaphenazole, phenytoin and (+/-)-warfarin was comparable in all livers, with values predicted from their Km or Ki for cytochrome P450TB determined in separate studies and a competitive inhibition model. Sulfaphenazole competitively inhibited diclofenac 4' hydroxylation (Ki = 0.11 +/- 0.08 microM, n = 3). Diclofenac 4'-hydroxylation is predominantly catalyzed by a cytochrome P450 isozyme of the CYP2C subfamily, most likely CYP2C9. This particular isozyme therefore appears to be responsible for the oxidation of polar acidic substances such as non-steroidal anti-inflammatory drugs from different chemical classes. It also constitutes a common site for drug interactions involving these compounds, as well as tolbutamide, phenytoin and warfarin. PMID- 8417278 TI - Transient forebrain ischemia alters acutely endothelin receptor density and immunoreactivity in gerbil brain. AB - Pharmacologic studies suggest that endothelin (ET) plays an important role in the pathophysiology of hemorrhagic and ischemic stroke. In the gerbil, transient forebrain ischemia (10 min) resulted in profound motor deficits and a 15% reduction in ET receptor density in the hippocampus at 60 min post-reperfusion. A significant 2-fold increase in forebrain immunoreactive ET accompanied the maximum post-ischemic decrease in ET receptor density. These results suggest that the synthesis and availability of ET are increased acutely in the forebrain following transient cerebral ischemia. PMID- 8417279 TI - Pregnancy can be established in superovulated adult rats treated with progesterone and an aromatase inhibitor. AB - An adult superovulated rat model has been developed and is characterized by high ovulation rates, early morphological degeneration of embryos, complete embryo loss within 48 hours of conception and elevated peripheral estradiol(E2)/progesterone(P4) ratios. In this study, three trials were conducted using the superovulated adult rat model. First, control naturally cycling rats were compared with superovulated rats supplemented with 1 mg P4 on days 0-3 of pregnancy. A sperm positive vaginal smear is designated as day 0 of pregnancy. The P4 treated rats demonstrated improved embryo retrieval on day 1 of pregnancy, continued embryo recovery with a decrease in normal morphologic characteristics of integrity on day 2, with nearly total embryo loss by day 3. On each day, P4 levels were elevated 2-3 times over control. The second trial compared 3 groups of rats, 1) naturally cycling, 2) superovulated unsupplemented and 3) superovulated rats supplemented with 1 mg P4/rat/day and the aromatase inhibitor, 4-hydroxyandrostenedione (4-OHA), 12.5 mg/rat/day. The superovulated unsupplemented rats had no embryo recovery after day 2 of pregnancy, while the P4 and 4-OHA treated rats showed a variable ability to maintain normally developing embryos through day 4 of pregnancy. E2 levels were elevated in both superovulated groups on days 1-4 of pregnancy as were P4 levels on days 2-4. The E2/P4 ratio was significantly lowered only on day 1 of pregnancy in the P4 and 4-OHA treated group. The third trial demonstrated implantation in 50% of the superovulated rats supplemented with P4 and 4-OHA. In conclusion, implantation in the superovulated adult rats can occur with P4 and 4-OHA supplementation, however, this biologic phenomenon could not be explained by obvious changes in peripheral E2 and P4 levels. PMID- 8417280 TI - Immunostimulatory factor(s) in the porcine uterine fluids. AB - The effects of porcine uterine fluid filtrate (PUF) on lymphocyte activity were examined. PUF was added to porcine lymphocyte cultures in the presence or absence of phytohemagglutinin (PHA), concanavalin A (Con A), or pokeweed mitogen (PWM). Lymphocyte response was determined by 3H-thymidine incorporation. Results indicated that PUF enhanced PHA-, Con A-, and PWM-induced lymphocyte mitogenesis by 222%, 207%, and 261%, respectively, as compared to control. The stimulatory effect of PUF was unlikely to be due to the presence of interleukin-2 in PUF, by evidence that PUF alone did not support cytotoxic T-lymphocyte proliferation, nor was the stimulatory effect due to microbial contamination, because treatment of PUF through PyroBind columns to remove endotoxin did not affect its activity. Incubation of PUF at 56C for 40 min did not alter the enhancement effect. Treatment of PUF with trypsin or pronase for up to 24 hr did not change its activity. In mixed lymphocytes reaction, PUF augmented 3H-thymidine uptake by 242%. The molecular (mol) mass of the immunostimulatory activity in PUF was examined by size-exclusion HPLC, Sephadex G-50, and G-15 gel filtration, and it was found to be 0.8 kDa and < 0.5 kDa. Extracts of porcine endometrium also enhanced PHA-induced lymphocyte mitogenesis, suggesting that the factor(s) may be of uterine origin. In conclusion, we have identified an factor(s) in PUF capable of stimulating lymphocyte mitogenesis and mixed lymphocyte reaction. PMID- 8417281 TI - Dichotomous cardiac and systemic vascular responses to cocaine in conscious rats. AB - This study examined the effects of cocaine on cardiac output in conscious freely moving rats. Although pressor responses were similar at all doses, 14 of 32 rats had consistent declines in cardiac output (> 15%) and greater increases in systemic vascular resistance after administration of cocaine (5 mg/Kg, i.v.). Procaine (10 mg/Kg i.v.) did not mimic this effect in either subgroup. We propose that a subpopulation of rats exists with an enhanced susceptibility to cocaine induced cardiac and systemic vascular alterations at higher doses. PMID- 8417282 TI - Identification and characterization of [125I]arginine vasopressin binding sites on human peripheral blood mononuclear cells. AB - Arginine vasopressin (AVP) is a nonapeptide that has been shown to be released from the posterior pituitary during stress. Although noted primarily for its hemodynamic and homeostatic properties, AVP also appears to have an effect on the immune system. It may modulate cellular immunity via its enhancement of the autologous mixed lymphocyte response (AMLR), an effect which we have demonstrated to occur over a wide dose range with a maximum at 10(-7) M. In this study, we examined the binding of [125I]AVP, and AVP analogues to human peripheral blood mononuclear cells (PBMC). AVP inhibited [125I]AVP (0.2 nM) binding on PBMC in a dose-dependent manner with maximal inhibition being reached at 10(-8) M. Specific [125I]AVP binding, as defined as that which could be displaced by 1 x 10(-6) M AVP, was saturable, time-dependent, and linear to cell concentration. Specific binding reached saturation at approximately 1000 pM in 45 minutes. From Scatchard analysis of saturation experiments it appeared to be a homogeneous population of binding sites with KD of approximately 0.5 nM and Bmax of approximately 7.6 fmole/8 x 10(6) cells, corresponding to approximately 527 binding sites/cell. There was a good correlation between AVP binding and cell number. AVP failed to dissociate completely from its binding sites in 60 minutes, perhaps because of the formation of a high-affinity ligand-binding site complex. From competitive binding studies with various AVP antagonists and analogues, it was found that the AVP binding site appeared to be V1-like. AVP binding occurred predominantly on B cells and macrophages. Having provided evidence for the existence of specific, high affinity, and saturable V1-like AVP binding sites, we suggest a potential modulatory role for AVP in the communication between the neuroendocrine and immune systems. PMID- 8417283 TI - The acquired immunodeficiency syndrome in Australia: incidence 1982-1991. AB - OBJECTIVE: To describe the incidence of the acquired immunodeficiency syndrome (AIDS) in Australia between 1982 and 1991. DESIGN: State and Territory Health Departments notified new diagnoses of AIDS to the National AIDS Registry. Information reported for each case included sex, date of birth, date of AIDS diagnosis, presumed mode of exposure to the human immunodeficiency virus (HIV), and illness(es) on which the diagnosis of AIDS was based. RESULTS: To the end of March 1992, 3,160 cases of AIDS were reported as having been diagnosed between 1982 and the end of 1991. The cumulative incidence per head of population was about twice as high in New South Wales as in Australia as a whole. Over 97% of cases were in men, of whom 91% were adults or adolescents reporting homosexual contact. In women, 40% of cases were acquired through receipt of blood, blood products or tissue. The annual incidence of AIDS rose sharply until about 1988, but the annual rates of increase slowed in subsequent years. This trend was also apparent in cases acquired through sexual contact between men. In other exposure groups, numbers of cases were much smaller and trends less apparent. However, there was no indication of a similar levelling in AIDS incidence, except among blood transfusion recipients, in whom incidence may be declining. CONCLUSION: Transmission of HIV among people with AIDS in Australia has been overwhelmingly attributed to sexual contact between men. The annual incidence of cases attributed to sexual contact between men appears to be stabilising. PMID- 8417284 TI - The epidemiology of HIV-1 infection in Victoria. The Victorian Collaborative Group on HIV and AIDS Surveillance (VCGHAS). AB - OBJECTIVE: To describe the epidemiology of infection with the human immunodeficiency virus type 1 (HIV-1) in Victoria from 1980 to 1991. DESIGN: Data on HIV-1 infection in Victoria, obtained through routine laboratory-based surveillance, were entered in a database. Missing information was sought by contacting the referring doctor where possible. SETTING: In Victoria, the acquired immunodeficiency syndrome (AIDS) is notifiable to Health Department Victoria by diagnosing doctors, and laboratories are required to notify new diagnoses of HIV-1 infection, without identifiers. All confirmatory testing for HIV-1 has taken place at the State HIV Reference Laboratory at Fairfield Hospital. MAIN OUTCOME MEASURES: Diagnoses of HIV-1 infection, as confirmed at the State HIV Reference Laboratory by western blot immunoassay, and notifications of AIDS to Health Department Victoria. RESULTS: Over six years the annual number of diagnoses of HIV-1 infection in Victoria remained constant despite a substantial increase in the number of tests performed. To the end of 1991, 2679 people had been diagnosed with HIV-1 infection, 686 of whom had developed AIDS. Information on exposure was available for 2379 (88.8%). Homosexual and bisexual men made up 75.5% (85.0% of those for whom exposure had been ascertained); 3.4% were female or heterosexual male injecting drug users; and 3.7% were heterosexuals with no history of injecting drug use. The latter two groups contributed 2.0% in 1985 to the proportion of all new diagnoses for which exposure was known, and 14.3% in 1991; for recipients of contaminated blood or blood products before 1985 this proportion fell from 12.4% to 1.0%. The cumulative incidence of HIV-1 diagnoses was highest in the age group 25-29 years, and 20% of all HIV-1 infected people were under 25 at the time of diagnosis. In 1991, 81 of the 311 people who had been diagnosed with HIV-1 infection had had previous negative or indeterminate results of tests; half of these had acquired infection in the previous year. CONCLUSION: Most HIV-1 infections in Victoria have been acquired through male homosexual contact, with a small but increasing proportion of diagnoses occurring in heterosexuals. Laboratory-based surveillance of voluntary testing, despite its limitations, has provided valuable information on the extent of the HIV-1 epidemic in Victoria. Surveillance of all HIV-1 test results and of seroconverters now supplements routine surveillance of HIV diagnoses and will ensure a more accurate picture of the epidemic in coming years. PMID- 8417285 TI - HIV risk factors and seroprevalence in surgical patients. AB - OBJECTIVE: To determine the prevalence of risk factors for human immunodeficiency virus (HIV) infection, and HIV-1 seroprevalence, in surgical patients. DESIGN: Prospective study in which consenting patients completed a questionnaire on factors potentially related to a higher risk of HIV infection, and underwent an HIV-1 antibody test. PATIENTS AND SETTING: Both elective and emergency surgical admissions between July and November 1990 at St Vincent's Hospital, Sydney. RESULTS: Of 1292 patients who were approached to participate, 27 had been previously diagnosed with HIV-1 infection and, of the remaining 1265, eight (0.63%) refused to answer the questionnaire and undergo a blood test, and 12 (0.95%) refused the blood test only. HIV-1 antibody testing was completed for 1171 study subjects. Twenty-six of the 807 men who answered the questionnaire had been previously diagnosed with HIV-1 infection. Of the remaining 781 male patients, 133 (17%) reported a history of homosexual contact, injecting drug use or blood transfusion, 132 (17%) had had a prior HIV-1 antibody test and three were newly diagnosed with HIV-1 infection as a result of the survey. Of 476 women who completed the questionnaire, one had been previously diagnosed with HIV-1 infection, and of the remaining 475, 59 (12.4%) reported injecting drug use or a blood transfusion and 72 (15%) had had a prior HIV-1 antibody test. No women were newly diagnosed with HIV-1 infection. Of patients reporting specific factors, the proportion who had had a prior HIV-1 antibody test varied from 62% for men reporting homosexual contact to 34% for recipients of a blood transfusion between 1980 and 1985. CONCLUSION: Although an appreciable proportion of surgical patients admitted to St Vincent's Hospital reported factors associated with a higher risk of HIV infection, the prevalence of undiagnosed HIV-1 infection is very low, particularly among patients reporting no such factors. PMID- 8417286 TI - Experience with 732 acute overdose patients admitted to an intensive care unit over six years. AB - OBJECTIVE: To determine the outcome in patients with acute overdose requiring admission to the Intensive Care Unit (ICU). DESIGN: Prospective survey of all overdose admissions to an ICU over a six-year period ending January 1991. SETTING: Tertiary referral adult teaching hospital. PATIENTS: 732 consecutive patients with acute overdose. OUTCOME MEASURES: Death rate, use and duration of mechanical ventilation, type of compound taken and compounds associated with a fatal outcome. RESULTS: The 732 patients represented 13.8% of all admissions and 6% of the available ICU bed-days. Comparison with all admissions to the Emergency Department for acute overdose over a 27-month period ending April 1990 indicated that 22% of these patients were admitted to the ICU. Among the patients admitted to the ICU, tricyclic antidepressants, benzodiazepines and alcohol were the most frequently used compounds. More than one compound had been taken by 46.8% of the patients. Mechanical ventilation was required in 79.5% of the patients and 14 (2%) died. CONCLUSIONS: Acute overdose is a common cause of admission to the ICU but has a mortality rate of only 2%. In contrast to the overdoses taken by survivors, patients taking fatal overdoses are more likely to have taken a large dose of a single drug, or a non-medicinal compound. PMID- 8417287 TI - The challenge: clinical diagnosis of HIV. AB - The diagnosis of HIV infection is missed too often, with significant costs to patients and the community. Could it be HIV? gives practical advice about the detection of unsuspected HIV infection following clinical presentation. Each section is written for doctors by colleagues with extensive experience in HIV medicine; many illustrative case histories are drawn from their personal records. The chronology of HIV-induced disease is a useful framework for interpreting the many possible clinical clues to HIV infection which, together with lifestyle clues, contribute to decision-making about the rational use of HIV testing. PMID- 8417288 TI - Benefits of early diagnosis of HIV infection. PMID- 8417289 TI - When do patients present with HIV infection? AB - A high proportion of people infected with HIV are not diagnosed until they have developed or are on the verge of developing AIDS. Ignorance of their risk of infection or fear of the implications of a diagnosis still play a part in their failure to have an HIV antibody test. Earlier diagnosis of HIV-induced disease is particularly important for this group, as is continuing education about the risks of infection and the value of early diagnosis. PMID- 8417290 TI - Outcome of missed diagnosis of HIV infection. AB - The potential benefits of early intervention in HIV infection with antiretroviral therapy and prophylaxis for opportunistic infections are well recognised. In contrast, data on the consequences of a missed diagnosis of HIV infection are limited. Two recently published studies address this issue. PMID- 8417291 TI - Lifestyle clues in the recognition of HIV infection. How to take a sexual history. PMID- 8417292 TI - HIV seroconversion illness. AB - HIV seroconversion is associated with characteristic clinical findings which reflect the first interaction of HIV with the host immune system. Recognition of the characteristic clinical illness associated with HIV seroconversion helps to establish an early diagnosis and obviates the need for further investigation or empiric therapy. It may also afford a unique opportunity to institute counselling to prevent the further spread of HIV in the community. PMID- 8417293 TI - The potential organ donor. AB - The source of most organs for transplantation is the brain-dead cadaveric donor. The realistic potential donor is one for whom mechanical ventilation and other support, to the point of brain death, is provided in the best interests of that patient, and not solely for the purpose of organ donation. Brain death secondary to spontaneous intracranial haemorrhage now exceeds traumatic head injury as a source of organ donors in Australia. There are now few medical exclusions to organ donation. Age limits rise constantly. No patient should be excluded without referral to the transplant coordinator. Adequate medical support of the potential donor is no more than should be provided to the severely brain-injured patient. Haemodynamic and other organ support are as logically appropriate as ventilation and must be continued until confirmation of brain death. Support should cease only if organ donation will not occur. Australian State laws define brain death as irreversible cessation of all function of the brain of the person, but do not dictate the methods of confirmation. The prospect of organ donation should not be raised until brain death is confirmed. The over-riding principle is that the family always has the right to be asked. There is no other way for the family's and patient's wishes about organ donation to be known and respected. It is vital that the person who will ask be experienced, competent and committed. A dedicated medical, nursing and allied health team providing care of the family throughout the period is essential. PMID- 8417294 TI - Acute compartment syndrome: an unusual presentation of gemfibrozil induced myositis. AB - OBJECTIVE: To present a case of gemfibrozil-induced myositis which precipitated an acute compartment syndrome. CLINICAL FEATURES: A 49-year-old woman with chronic renal failure was given gemfibrozil for hyperlipidaemia. She developed myositis in the sixth week of therapy. Her symptoms initially persisted despite withdrawal of gemfibrozil and she developed an acute compartment syndrome. INTERVENTION AND OUTCOME: Emergency fasciotomy was performed. She gradually improved and complete recovery occurred eight weeks after cessation of gemfibrozil therapy. CONCLUSION: Gemfibrozil may induce a local myositis which, in our patient, precipitated an acute compartment syndrome. One should be alert to symptoms of possible drug-induced myositis in patients receiving gemfibrozil. Extreme caution should be exercised in its use in patients with impaired renal function. PMID- 8417295 TI - Deep venous thrombosis caused by femoral exostosis. AB - OBJECTIVE: To present the first case of deep venous thrombosis caused by femoral exostosis reported in Australia. CLINICAL FEATURES: An 11-year-old prepubertal Caucasian girl had a two-year history of a posterior femoral exostosis. She then presented with a deep venous thrombosis 24 hours after riding a horse for the first time. The deep venous thrombosis was diagnosed by Doppler ultrasound, which showed an intimate relationship between the femoral exostosis and the femoral vein. Coagulation abnormalities were excluded. INTERVENTION AND OUTCOME: The patient was given anticoagulation therapy with heparin intravenously and warfarin orally. The popliteal vein recanalised within two days. The exostosis was excised 10 weeks after initial presentation, with warfarin being continued for four weeks postoperatively. Two months after excision the patient was symptom free. CONCLUSIONS: It is possible for venous compression by an exostosis to result in thrombosis in a patient with no underlying coagulation abnormality. Palpation and plain radiography of the region will demonstrate the exostosis; ultrasound is the next investigation of choice. PMID- 8417296 TI - Magnetic resonance angiography. AB - OBJECTIVE: To outline the physical principles of magnetic resonance angiography (MRA), major current MRA techniques, clinical indications and some of the limitations of MRA. The rapidly changing nature of MRA techniques is stressed. DATA SOURCES AND DATA EXTRACTION: The MRA literature for the last eight years is reviewed with particular attention to the changing clinical role of MRA. Details of specific MRA techniques are omitted for simplicity and because of their lack of direct relevance to the clinicians requesting MRA examinations. Data concerning the clinical role of MRA techniques are extracted from the literature and combined with our own clinical experience. DATA SYNTHESIS: While stressing the rapidly changing nature of this new investigational technique, we find that MRA has a number of current roles in patient management. CONCLUSIONS: MRA is currently a useful investigation and will in all likelihood become more useful in the future as the technique improves further and becomes more available. PMID- 8417297 TI - A numbers game. PMID- 8417298 TI - Assessing impairment of the back, neck and pelvis. PMID- 8417299 TI - Assessing impairment of the back, neck and pelvis. PMID- 8417300 TI - Assessing impairment of the back, neck and pelvis. PMID- 8417301 TI - Are Papanicolaou recalls worthwhile? PMID- 8417302 TI - The cochlear implant and the deaf community. PMID- 8417303 TI - Heartburn--lifting the veil of mythology. PMID- 8417304 TI - Some characteristics of blood shed into the Solcotrans postoperative orthopaedic drainage/reinfusion system. PMID- 8417305 TI - The excimer laser in Australia. PMID- 8417306 TI - Spontaneously remitting disease. PMID- 8417307 TI - Verminous asthma. PMID- 8417308 TI - Patient confidentiality. PMID- 8417309 TI - Occupational skin disease in New South Wales. PMID- 8417310 TI - The heterogeneity of homosexuality. PMID- 8417311 TI - Whiplash in Australia: illness or injury? PMID- 8417312 TI - Whiplash in Australia: illness or injury? PMID- 8417313 TI - Balkan ophthalmic neuralgia: an unusual war-related facial pain. PMID- 8417314 TI - Laparoscopic surgery: some unresolved issues. PMID- 8417315 TI - Occupational deaths. PMID- 8417316 TI - Whiplash in Australia: illness or injury? PMID- 8417317 TI - Interactions among the subunits of the G protein involved in Saccharomyces cerevisiae mating. AB - The SCG1 (GPA1), STE4, and STE18 genes of Saccharomyces cerevisiae encode mating pathway components whose amino acid sequences are similar to those of the alpha, beta, and gamma subunits, respectively, of mammalian G proteins. Genetic evidence suggests that the STE4 and STE18 gene products interact. The mating defects of a set of ste4 mutants were partially suppressed by the overexpression of STE18, and, moreover, a combination of partially defective ste4 and ste18 alleles created a totally sterile phenotype, whereas such synthetic sterility was not observed when the ste18 allele was combined with a weakly sterile ste11 allele. Others have provided genetic evidence consistent with an interaction between the SCG1 (GPA1) and STE4 gene products. We have examined the physical interactions of these subunits by using an in vivo protein association assay. The STE4 and STE18 gene products associated with each other, and this association was disrupted by a mutation in the STE4 gene product whose phenotype was partially suppressed by overexpression of STE18. The STE4 and SCG1 (GPA1) gene products also interacted in the assay, whereas we detected no association of the SCG1 (GPA1) and STE18 gene products. PMID- 8417318 TI - A rat gene with sequence homology to the Drosophila gene hairy is rapidly induced by growth factors known to influence neuronal differentiation. AB - Several genes encoding transcription factors with a helix-loop-helix (HLH) motif are involved in the early process of neural development in Drosophila spp. We report the isolation from the rat a homolog of one of these genes, called hairy. The rat-hairy-like (RHL) gene is expressed early during embryogenesis. In contrast to the restricted expression of hairy mRNA in Drosophila spp., however, the mRNA encoded by RHL is detectable in all tissues examined. Stimulation of PC12 pheochromocytoma cells by nerve growth factor, basis fibroblast growth factor, or epidermal growth factor or of Rat-1 fibroblasts by epidermal growth factor causes a rapid and transient induction of the RHL gene. Thus, RHL acts as an immediate-early gene that can potentially transduce growth factor signals during the development of the mammalian embryo. PMID- 8417319 TI - Gene RRN4 in Saccharomyces cerevisiae encodes the A12.2 subunit of RNA polymerase I and is essential only at high temperatures. AB - We have previously isolated mutants of Saccharomyces cerevisiae that are primarily defective in transcription of 35S rRNA genes by RNA polymerase I and have identified genes (RRN1 to RRN9) involved in this process. We have now cloned the RRN4 gene by complementation of the temperature-sensitive phenotype of the rrn4-1 mutant and have determined its complete nucleotide sequence. The following results demonstrate that the RRN4 gene encodes the A12.2 subunit of RNA polymerase I. First, RRN4 protein expressed in Escherichia coli reacted with a specific antiserum against A12.2. Second, amino acid sequences of three tryptic peptides obtained from A12.2 were determined, and these sequences are found in the deduced amino acid sequence of the RRN4 protein. The amino acid sequence of the RRN4 protein (A12.2) is similar to that of the RPB9 (B12.6) subunit of yeast RNA polymerase II; the similarity includes the presence of two putative zinc binding domains. Thus, A12.2 is a homolog of B12.6. We propose to rename the RRN4 gene RPA12. Deletion of RPA12 produces cells that are heat but not cold sensitive for growth. We have found that in such null mutants growing at permissive temperatures, the cellular concentration of A190, the largest subunit of RNA polymerase I, is lower than in the wild type. In addition, the temperature sensitive phenotype of the rpa12 null mutants can be partially suppressed by RPA190 (the gene for A190) on multicopy plasmids. These results suggest that A12.2 plays a role in the assembly of A190 into a stable polymerase I structure. PMID- 8417320 TI - Identification of amino acids essential for DNA binding and dimerization in p67SRF: implications for a novel DNA-binding motif. AB - The serum response factor (p67SRF) binds to a palindromic sequence in the c-fos serum response element (SRE). A second protein, p62TCF binds in conjunction with p67SRF to form a ternary complex, and it is through this complex that growth factor-induced transcriptional activation of c-fos is thought to take place. A 90 amino-acid peptide, coreSRF, is capable for dimerizing, binding DNA, and recruiting p62TCF. By using extensive site-directed mutagenesis we have investigated the role of individual coreSRF amino acids in DNA binding. Mutant phenotypes were defined by gel retardation and cross-linking analyses. Our results have identified residues essential for either DNA binding or dimerization. Three essential basic amino acids whose conservative mutation severely reduced DNA binding were identified. Evidence which is consistent with these residues being on the face of a DNA binding alpha-helix is presented. A phenylalanine residue and a hexameric hydrophobic box are identified as essential for dimerization. The amino acid phasing is consistent with the dimerization interface being presented as a continuous region on a beta-strand. A putative second alpha-helix acts as a linker between these two regions. This study indicates that p67SRF is a member of a protein family which, in common with many DNA binding proteins, utilize an alpha-helix for DNA binding. However, this alpha helix is contained within a novel domain structure. PMID- 8417321 TI - RNA B is the major nucleolar trimethylguanosine-capped small nuclear RNA associated with fibrillarin and pre-rRNAs in Trypanosoma brucei. AB - RNA B is one of three abundant trimethylguanosine-capped U small nuclear RNAs (snRNAs) of Trypanosoma brucei which is not strongly identified with other U snRNAs by sequence homology. We show here that RNA B is a highly diverged U3 snRNA homolog likely involved in pre-rRNA processing. Sequence identity between RNA B and U3 snRNAs is limited; only two of four boxes of homology conserved between U3 snRNAs are obvious in RNA B. These are the box A homology, specific for U3 snRNAs, and the box C homology, common to nucleolar snRNAs and required for association with the nucleolar protein, fibrillarin. A 35-kDa T. brucei fibrillarin homolog was identified by using an anti-Physarum fibrillarin monoclonal antibody. RNA B and fibrillarin were localized in nucleolar fractions of the nucleus which contained pre-rRNAs and did not contain nucleoplasmic snRNAs. Fibrillarin and RNA B were precipitated by scleroderma patient serum S4, which reacts with fibrillarins from diverse organisms; RNA B was the only trimethylguanosine-capped RNA precipitated. Furthermore, RNA B sedimented with pre-rRNAs in nondenaturing sucrose gradients, similarly to U3 and other nucleolar snRNAs, suggesting that RNA B is hydrogen bonded to rRNA intermediates and might be involved in their processing. PMID- 8417322 TI - Ras activation by insulin and epidermal growth factor through enhanced exchange of guanine nucleotides on p21ras. AB - A number of growth factors, including insulin and epidermal growth factor (EGF), induce accumulation of the GTP-bound form of p21ras. This accumulation could be caused either by an increase in guanine nucleotide exchange on p21ras or by a decrease in the GTPase activity of p21ras. To investigate whether insulin and EGF affect nucleotide exchange on p21ras, we measured binding of [alpha-32P]GTP to p21ras in cells permeabilized with streptolysin O. For this purpose, we used a cell line which expressed elevated levels of p21 H-ras and which was highly responsive to insulin and EGF. Stimulation with insulin or EGF resulted in an increase in the rate of nucleotide binding to p21ras. To determine whether this increased binding rate is due to the activation of a guanine nucleotide exchange factor, we made use of the inhibitory properties of a dominant negative mutant of p21ras, p21ras (Asn-17). Activation of p21ras by insulin and EGF in intact cells was abolished in cells infected with a recombinant vaccinia virus expressing p21ras (Asn-17). In addition, the enhanced nucleotide binding to p21ras in response to insulin and EGF in permeabilized cells was blocked upon expression of p21ras (Asn-17). From these data, we conclude that the activation of a guanine nucleotide exchange factor is involved in insulin- and EGF-induced activation of p21ras. PMID- 8417323 TI - An abundant high-mobility-group-like protein is targeted to micronuclei in a cell cycle-dependent and developmentally regulated fashion in Tetrahymena thermophila. AB - In this report, we have demonstrated for the first time that an abundant high mobility-group (HMG)-like protein, HMG B, previously thought to be specific to macronuclei in Tetrahymena thermophila, is also present in micronuclei. Biochemical data document the fact that HMG B is extremely labile in micronuclei. Unless extreme precautions are taken during the isolation of nuclei (addition of 1% formaldehyde to the nucleus isolation buffer), HMG B is not detected in micronuclei. Using polyclonal antibodies highly selective for HMG B, immunoblotting and immunofluorescence analyses show that the presence of HMG B in micronuclei is dynamic, correlating well with known periods of micronuclear DNA replication. This is the case not only during the vegetative cell cycle but also during early stages of the sexual cycle, conjugation, when the presence of HMG B in micronuclei is also closely correlated with meiotic DNA recombination and repair. Since micronuclei are transcriptionally inactive during vegetative growth, our data lend support to the idea that HMG B does not function exclusively in the establishment of transcriptionally competent chromatin. However, micronuclei are transcriptionally active during early stages of conjugation. Evidence that HMG B is strongly synthesized and deposited into micronuclei during this stage is presented. Therefore, it is tempting to suggest that HMG B may play an important role in remodeling micronuclear chromatin into an "active," more open configuration. We favor a model wherein HMG B, like other abundant, low-specificity HMG box-containing proteins, functions to wrap DNA, presumably modulating higher-order chromatin structure for a broad range of biological processes, including transcription and replication. PMID- 8417324 TI - Isolation of RRM-type RNA-binding protein genes and the analysis of their relatedness by using a numerical approach. AB - Proteins with RNA recognition motifs (RRMs) have important roles in a great many aspects of RNA metabolism. However, this family has yet to be systematically studied in any single organism. In order to investigate the size of the RRM gene family in Drosophila melanogaster and to clone members of this family, we used a polymerase chain reaction (PCR) with highly degenerate oligonucleotides to amplify DNA fragments between the RNP-1 and RNP-2 consensus sequences of the RRM proteins. Cloning and sequencing of 124 PCR products revealed 12 different RRM sequences (RRM1 to RRM12). When PCR products were used as probes in genomic Southern and Northern (RNA) analyses, 16 restriction fragments and 25 transcripts, respectively, were detected. Since the combinations of nucleotide sequences represented in the PCR primers correspond to only 4% of the RRM sequences inferred to be possible from known RRM sequences, we estimate the size of the RRM gene family in the order of three hundred genes in flies. In order to gain insight into the possible functions of the genes encoding the RRMs, we analyzed the sequence similarities between the 12 RRMs and 62 RRM sequences of known proteins. This analysis showed that the RRMs of functionally related proteins have similar sequences and are clustered together in the RRM gene tree. On the basis of this observation, the RRMs can be divided into three groups: a heterogeneous nuclear ribonucleoprotein type, a splicing regulator type, and a development-specific factor type. This result suggests that we have isolated good candidates for both housekeeping and developmentally important genes involved in RNA metabolism. PMID- 8417325 TI - A transcriptional switch between the Pig-1 and Sgs-4 genes of Drosophila melanogaster. AB - Pig-1 and Sgs-4 are a pair of closely linked and divergently transcribed Drosophila melanogaster genes, which are both expressed in larval salivary glands but at different times during development. While Sgs-4 is expressed at high levels only at the end of the third instar, Pig-1 exhibits a major peak of expression during late second and early third instar. Thus, Pig-1 expression declines as Sgs-4 expression is induced. In this paper, we show that three adjacent elements located within the short region between these genes can account for the switch from Pig-1 to Sgs-4 expression. A 170-bp segment acts as an enhancer to direct Sgs-4 expression in late-third-instar salivary glands. A 64-bp sequence located just upstream from the enhancer can modify its temporal specificity so that it works throughout the third instar. Expression induced at mid-third instar by a combination of these two elements can be repressed by a negative regulatory sequence located still further upstream. We present evidence suggesting that the changing interactions between these regulatory elements and the Sgs-4 and Pig-1 promoters lead to the correct pattern of expression of the two genes. PMID- 8417326 TI - Two domains of ISGF3 gamma that mediate protein-DNA and protein-protein interactions during transcription factor assembly contribute to DNA-binding specificity. AB - Alpha interferon (IFN-alpha) induces the transcription of a large set of genes through activation of multimeric transcription factor ISGF3. This factor can be dissociated into two protein components, termed ISGF3 gamma and ISGF3 alpha. ISGF3 gamma is a 48-kDa protein related at the amino terminus to members of the IFN-regulatory factor (IRF) and Myb families of DNA-binding proteins; ISGF3 alpha consists of three polypeptides of 84, 91, and 113 kDa that self-assemble to form an activated component in response to IFN-alpha. DNA-binding studies indicated that ISGF3 gamma binds DNA alone, recognizing the IFN-stimulated response element, while the ISGF3 alpha polypeptides alone display no specific interactions with DNA. A complex between ISGF3 gamma and activated ISGF3 alpha binds the IFN-stimulated response element with much greater affinity than does the 48-kDa ISGF3 gamma protein alone. The DNA-binding domain of ISGF3 gamma and regions responsible for protein-protein interaction with ISGF3 alpha were identified by using deleted forms of ISGF3 gamma expressed in vitro. The amino terminal region of ISGF3 gamma homologous to the IRF and Myb proteins was sufficient for interaction with DNA and displayed the binding specificity of the intact protein; phosphorylation of this region was necessary for activity. A second region of 160 amino acids separated from the DNA-binding domain by over 100 amino acids contained a domain capable of associating with ISGF3 alpha and was sufficient to confer specific ISGF3 alpha interaction to a heterologous protein. Interaction of the ISGF3 alpha component with the protein interaction domain of ISGF3 gamma altered the DNA-binding specificity of the resulting complex, suggesting that one or more of the ISGF3 alpha polypeptides make base specific contacts with DNA. This interaction defines a mechanism through which IRF-like proteins complexed with regulatory components can display novel DNA binding specificities. PMID- 8417327 TI - A Drosophila homolog of bovine smg p25a GDP dissociation inhibitor undergoes a shift in isoelectric point in the developmental mutant quartet. AB - The Drosophila developmental mutation quartet causes late larval lethality and small imaginal discs and, when expressed in the adult female, has a lethal effect on early embryogenesis. These developmental defects are associated with mitotic defects, which include a low mitotic index in larval brains and incomplete separation of chromosomes in mitosis in the early embryo. quartet mutations also have a biochemical effect, i.e., a basic shift in isoelectric point in three proteins. We have purified one of these proteins, raised an antibody to it, and isolated and sequenced its cDNA. At the amino acid level, the sequence shows 68% identity and 81% similarity to bovine smg p25a GDP dissociation inhibitor (GDI), a regulator of ras-like small GTPases of the rab/SEC4/YPT1 subfamily. The correlation between a basic shift in isoelectric point in Drosophila GDI in quartet mutant tissue and the quartet developmental phenotype raises the possibility that a posttranslational modification of GDI is necessary for its function and that GDI function is essential for development. PMID- 8417328 TI - Molecular basis for developmental changes in interleukin-2 gene inducibility. AB - At least three stages in the intrathymic development of pre-T cells are demarcated by differences in the competence to express the interleukin-2 (IL-2) gene as an acute response to stimulation. IL-2 inducibility appears to be acquired relatively early, prior to T-cell receptor (TcR) gene rearrangement. It is then abrogated during the stage when cells are subject to positive and negative selection, i.e., the fate determination processes that select cells for maturation or death. IL-2 inducibility finally reappears in mature classes of thymocytes that have undergone positive selection. To provide a basis for a molecular explanation of these developmental transitions, we have examined the representation in different thymocyte subsets of a set of DNA-binding proteins implicated in IL-2 gene regulation. As the DNA-binding activities of many factors are elicited only by inductive stimuli, the cells were cultured in the presence or absence of the calcium ionophore A23187 and phorbol ester. Our results separate these factors into four regulatory classes: (i) constitutive factors, such as Oct-1 and probably Sp1, that are expressed in thymocytes at all stages; (ii) inducible factors, such as NF-kappa B and complexes binding to the region of a CD28 response element, that can be activated in all thymocytes, including those cells (CD4+ CD8+ TcRlow) that can undergo selection; (iii) inducible factors, such as NF-AT and AP-1, that can be activated in mature (CD4+ CD8- TcRhigh) and immature (CD4- CD8- TcR-) thymocytes alike but not in the transitional stages when the cells (CD4+ CD8+ TcRlow) are subject to selection; and (iv) a factor containing CREB, which can be activated in thymocytes of all developmental stages by culture but does not require specific induction. These results verify that inducible transcription factors are targets of intrathymic developmental change. They also identify NF-AT and AP-1 as factors that are particularly sensitive to the mechanism altering thymocyte responses during the stages when thymocytes may undergo positive and negative selection. PMID- 8417329 TI - Localization of small heat shock proteins to the higher plant endomembrane system. AB - Three related gene families of low-molecular-weight (LMW) heat shock proteins (HSPs) have been characterized in plants. We describe a fourth LMW HSP family, represented by PsHSP22.7 from Pisum sativum and GmHSP22.0 from Glycine max, and demonstrate that this family of proteins is endomembrane localized. PsHSP22.7 and GmHSP22.0 are 76.7% identical at the amino acid level. Both proteins have amino terminal signal peptides and carboxyl-terminal sequences characteristic of endoplasmic reticulum (ER) retention signals. The two proteins closely resemble class I cytoplasmic LMW HSPs, suggesting that they evolved from the cytoplasmic proteins through the addition of the signal peptide and ER retention motif. The endomembrane localization of these proteins was confirmed by cell fractionation. The polypeptide product of PsHSP22.7 mRNA was processed to a smaller-M(r) form by canine pancreatic microsomes; in vivo, GmHSP22.0 polysomal mRNA was found to be predominantly membrane bound. In vitro-processed PsHSP22.7 corresponded in mass and pI to one of two proteins detected in ER fractions from heat-stressed plants by using anti-PsHSP22.7 antibodies. Like other LMW HSPs, PsHSP22.7 was observed in higher-molecular-weight structures with apparent masses of between 80 and 240 kDa. The results reported here indicate that members of this new class of LMW HSPs are most likely resident ER proteins and may be similar in function to related LMW HSPs in the cytoplasm. Along with the HSP90 and HSP70 classes of HSPs, this is the third category of HSPs localized to the ER. PMID- 8417330 TI - Identification of cis and trans components of a novel heat shock stress regulatory pathway in Saccharomyces cerevisiae. AB - The stress-responsive DDR2 gene (previously called DDRA2) of Saccharomyces cerevisiae is transcribed at elevated levels following stress caused by heat shock or DNA damage. Previously, we identified a 51-bp promoter fragment, oligo31/32, which conferred heat shock inducibility on the heterologous CYC1-lacZ reporter gene in S. cerevisiae (N. Kobayashi and K. McEntee, Proc. Natl. Acad. Sci. USA 87:6550-6554, 1990). Using a series of synthetic oligonucleotides, we have identified a pentanucleotide, CCCCT (C4T), as an essential component of this stress response sequence. This element is not a binding site for the well characterized heat shock transcription factor which recognizes a distinct cis acting heat shock element in the promoters of many heat shock genes. Here we demonstrate the ability of oligonucleotides containing the C4T sequence to confer heat shock inducibility on the reporter gene and show that the presence of two such elements produces more than additive effects on induction. Gel retardation experiments have been used to demonstrate specific complex formation between C4T containing fragments and one or more yeast proteins. Formation of these complexes was not competed by fragments containing mutations in the C4T sequence nor by heat shock element-containing competitor DNAs. Fragments containing the C4T element bound to a single 140-kDa polypeptide, distinct from heat shock transcription factors in yeast crude extracts. These experiments identify key cis and trans-acting components of a novel heat shock stress response pathway in S. cerevisiae. PMID- 8417331 TI - Characterization of HIR1 and HIR2, two genes required for regulation of histone gene transcription in Saccharomyces cerevisiae. AB - The products of the HIR1 and HIR2 genes have been defined genetically as repressors of histone gene transcription in S. cerevisiae. A mutation in either gene affects cell cycle regulation of three of the four histone gene loci; transcription of these loci occurs throughout the cell cycle and is no longer repressed in response to the inhibition of DNA replication. The same mutations also eliminate autogenous regulation of the HTA1-HTB1 locus by histones H2A and H2B. The HIR1 and HIR2 genes have been isolated, and their roles in the transcriptional regulation of the HTA1-HTB1 locus have been characterized. Neither gene encodes an essential protein, and null alleles derepress HTA1-HTB1 transcription. Both HIR genes are expressed constitutively under conditions that lead to repression or derepression of the HTA1 gene, and neither gene regulates the expression of the other. The sequence of the HIR1 gene predicts an 88-kDa protein with three repeats of a motif found in the G beta subunit of retinal transducin and in a yeast transcriptional repressor, Tup1. The sequence of the HIR2 gene predicts a protein of 98 kDa. Both gene products contain nuclear targeting signals, and the Hir2 protein is localized in the nucleus. PMID- 8417332 TI - Direct selection for mutations affecting specific splice sites in a hamster dihydrofolate reductase minigene. AB - A Chinese hamster cell line containing an extra exon 2 (50 bp) inserted into a single intron of a dihydrofolate reductase (dhfr) minigene was constructed. The extra exon 2 was efficiently spliced into the RNA, resulting in an mRNA that is incapable of coding for the DHFR enzyme. Mutations that decreased splicing of this extra exon 2 caused it to be skipped and so produced normal dhfr mRNA. In contrast to the parental cell line, the splicing mutants display a DHFR-positive growth phenotype. Splicing mutants were isolated from this cell line after treatment with four different mutagens (racemic benzo[c]phenanthrene diol epoxide, ethyl methanesulfonate, ethyl nitrosourea, and UV irradiation). By polymerase chain reaction amplification and direct DNA sequencing, we determined the base changes in 66 mutants. Each of the mutagens generated highly specific base changes. All mutations were single-base substitutions and comprised 24 different changes distributed over 16 positions. Most of the mutations were within the consensus sequences at the exon 2 splice donor, acceptor, and branch sites. The RNA splicing patterns in the mutants were analyzed by quantitative reverse transcription-polymerase chain reaction. The recruitment of cryptic sites was rarely seen; simple exon skipping was the predominant mutant phenotype. The wide variety of mutations that produced exon skipping suggests that this phenotype is the typical consequence of splice site damage and supports the exon definition model of splice site selection. A few mutations were located outside the consensus sequences, in the exon or between the branch point and the polypyrimidine tract, identifying additional positions that play a role in splice site definition. That most of these 66 mutations fell within consensus sequences in this near-saturation mutagenesis suggests that splicing signals beyond the consensus may consist of robust RNA structures. PMID- 8417333 TI - The mdm-2 oncogene can overcome wild-type p53 suppression of transformed cell growth. AB - Expression of a p53-associated protein, Mdm-2 (murine double minute-2), can inhibit p53-mediated transactivation. In this study, overexpression of the Mdm-2 protein was found to result in the immortalization of primary rat embryo fibroblasts (REFs) and, in conjunction with an activated ras gene, in the transformation of REFs. The effect of wild-type p53 on the transforming properties of mdm-2 was determined by transfecting REFs with ras, mdm-2, and normal p53 genes. Transfection with ras plus mdm-2 plus wild-type p53 resulted in a 50% reduction in the number of transformed foci (relative to the level for ras plus mdm-2); however, more than half (9 of 17) of the cell lines derived from these foci expressed low levels of a murine p53 protein with the characteristics of a wild-type p53. These results are in contrast to previous studies which demonstrated that even minimal levels of wild-type p53 are not tolerated in cells transformed by ras plus myc, E1A, or mutant p53. The mdm-2 oncogene can overcome the previously demonstrated growth-suppressive properties of p53. PMID- 8417334 TI - Transforming growth factor alpha dramatically enhances oncogene-induced carcinogenesis in transgenic mouse pancreas and liver. AB - To characterize the effect(s) of transforming growth factor alpha (TGF alpha) during multistage carcinogenesis, we examined tumor development in pancreas and liver of transgenic mice that coexpressed TGF alpha with either viral (simian virus 40 T antigens [TAg]) or cellular (c-myc) oncogenes. In pancreas, TGF alpha itself was not oncogenic, but it nevertheless dramatically accelerated growth of tumors induced by either oncogene alone, thereby reducing the host life span up to 60%. Coexpression of TGF alpha and TAg produced an early synergistic growth response in the entire pancreas together with the more rapid appearance of preneoplastic foci. Coexpression of TGF alpha and c-myc also accelerated tumor growth in situ and produced transplantable acinar cell carcinomas whose rate of growth was TGF alpha dependent. In liver, expression of TGF alpha alone increased the incidence of hepatic cancer in aged mice. However, coexpression of TGF alpha with c-myc or TAg markedly reduced tumor latency and accelerated tumor growth. Significantly, expression of the TGF alpha and myc transgenes in hepatic tumors was induced up to 20-fold relative to expression in surrounding nonneoplastic liver, suggesting that high-level overexpression of these proteins acts as a major stimulus for tumor development. Finally, in both pancreas and liver, combined expression of TGF alpha and c-myc produced tumors with a more malignant (less differentiated) appearance than did expression of c-myc alone, consistent with an influence of TGF alpha upon the morphological character of c-myc-induced tumor progression. These findings demonstrate the importance of TGF alpha expression during multistage carcinogenesis in vivo and point to a major role for this growth factor as a potent stimulator of tumor growth. PMID- 8417335 TI - An essential yeast gene with homology to the exonuclease-encoding XRN1/KEM1 gene also encodes a protein with exoribonuclease activity. AB - An essential gene, designated HKE1/RAT1, has been isolated from the yeast Saccharomyces cerevisiae and characterized. The gene encodes a protein of 116 kDa (p116) and has significant homology to another yeast gene (XRN1/KEM1) encoding a related protein (p175) with 5'-->3' exonuclease activity as well as activities involving chromosomal DNA pairing and mechanics. Preliminary analysis of an hke1ts mutant reveals a precipitous decline in the translation of mRNA at the nonpermissive temperature. Sporulation of heterozygous HKE1/hke1::URA3 diploids reveals that this gene, unlike the highly related XRN1/KEM1 gene, is essential for cell viability. Overexpression of the homologous gene product, p175, failed to rescue cells lacking a functional p116. In vitro studies demonstrate that p116 is a protein with 5'-->3' exoribonuclease activity, a major activity of the related p175. An immunoreactive RNase activity of 116 kDa is abolished with antiserum against p116. Both the level of this protein and the RNase activity correlate with HKE1 gene dosage. The RNase activity purifies coincidentally with a previously described 116-kDa RNase having 5'-->3' exoribonuclease activity. PMID- 8417336 TI - Timing of molecular events in meiosis in Saccharomyces cerevisiae: stable heteroduplex DNA is formed late in meiotic prophase. AB - To better understand the means by which chromosomes pair and recombine during meiosis, we have determined the time of appearance of heteroduplex DNA relative to the times of appearance of double-strand DNA breaks and of mature recombined molecules. Site-specific double-strand breaks appeared early in meiosis and were formed and repaired with a timing consistent with a role for breaks as initiators of recombination. Heteroduplex-containing molecules appeared about 1 h after double-strand breaks and were followed shortly by crossover products and the first meiotic nuclear division. We conclude that parental chromosomes are stably joined in heteroduplex-containing structures late in meiotic prophase and that these structures are rapidly resolved to yield mature crossover products. If the chromosome pairing and synapsis observed earlier in meiotic prophase is mediated by formation of biparental DNA structures, these structures most likely either contain regions of non-Watson-Crick base pairs or contain regions of heteroduplex DNA that either are very short or dissociate during DNA purification. Two loci were examined in this study: the normal ARG4 locus, and an artificial locus consisting of an arg4-containing plasmid inserted at MAT. Remarkably, sequences in the ARG4 promoter that suffered double-strand cleavage at the normal ARG4 locus were not cut at significant levels when present at MAT::arg4. These results indicate that the formation of double-strand breaks during meiosis does not simply involve the specific recognition and cleavage of a short nucleotide sequence. PMID- 8417337 TI - Sequence-specific transcriptional activation by Myc and repression by Max. AB - The c-Myc oncoprotein, which is required for cellular proliferation, resembles in its structure a growing number of transcription factors. However, the mechanism of its action in vivo is not yet clear. The discovery of the specific cognate DNA binding site for Myc and its specific heterodimerization partner, Max, enabled the use of direct experiments to elucidate how Myc functions in vivo and how this function is modulated by Max. Here we demonstrate that exogenously expressed Myc is capable of activating transcription in vivo through its specific DNA-binding site. Moreover, transcriptional activation by Myc is dependent on the basic region, the integrity of the helix-loop-helix and leucine zipper dimerization motifs located in the carboxy-terminal portion of the protein, and the regions in the amino terminus conserved among Myc family proteins. In contrast to Myc, exogenously expressed Max elicited transcriptional repression and blocked transcriptional activation by Myc through the same DNA-binding site. Our results suggest a functional antagonism between Myc and Max which is mediated by their relative levels in the cells. A model for the activity of Myc and Max in vivo is presented. PMID- 8417338 TI - The effect on chromosome stability of deleting replication origins. AB - The observed spacing between chromosomal DNA replication origins in Saccharomyces cerevisiae is at least four times shorter than should be necessary to ensure complete replication of chromosomal DNA during the S phase. To test whether all replication origins are required for normal chromosome stability, the loss rates of derivatives of chromosome III from which one or more origins had been deleted were measured. In the case of a 61-kb circular derivative of the chromosome that has two highly active origins and one origin that initiates only 10 to 20% of the time, deletion of either highly active origin increased its rate of loss two- to fourfold. Deletion of both highly active origins caused the ring chromosome to be lost in approximately 20% of cell divisions. This very high rate of loss demonstrates that there are no efficient cryptic origins on the ring chromosome that are capable of ensuring its replication in the absence of the origins that are normally used. Deletion of the same two origins from the full-length chromosome III, which contains more than six replication origins, had no effect on its rate of loss. These results suggest that the increase in the rate of loss of the small circular chromosome from which a single highly active origin was deleted was caused by the failure of the remaining highly active origin to initiate replication in a small fraction (approximately 0.003) of cell cycles. PMID- 8417339 TI - Direct interaction of the tau 1 transactivation domain of the human glucocorticoid receptor with the basal transcriptional machinery. AB - We have used a yeast (Saccharomyces cerevisiae) cell free transcription system to study protein-protein interactions involving the tau 1 transactivation domain of the human glucocorticoid receptor that are important for transcriptional transactivation by the receptor. Purified tau 1 specifically inhibited transcription from a basal promoter derived from the CYC1 gene and from the adenovirus 2 major late core promoter in a concentration-dependent manner. This inhibition or squelching was correlated with the transactivation activity of tau 1. Recombinant yeast TATA-binding protein (yTFIID), although active in vitro, did not specifically reverse the inhibitory effect of tau 1. In addition, no specific interaction between tau 1 and yTFIID could be shown in vitro by affinity chromatography. Taken together, these results indicate that the tau 1 transactivation domain of the human glucocorticoid receptor interacts directly with the general transcriptional apparatus through some target protein(s) that is distinct from the TATA-binding factor. Furthermore, this assay can be used to identify interacting factors, since after phosphocellulose chromatography of a whole-cell yeast extract, a fraction that contained an activity which selectively counteracted the squelching effect of tau 1 was found. PMID- 8417340 TI - Developmental regulation and tissue distribution of the liver transcription factor LFB1 (HNF1) in Xenopus laevis. AB - The transcription factor LFB1 (HNF1) was initially identified as a regulator of liver-specific gene expression in mammals. It interacts with the promoter element HP1, which is functionally conserved between mammals and amphibians, suggesting that a homologous factor, XLFB1, also exists in Xenopus laevis. To study the role of LFB1 in early development, we isolated two groups of cDNAs coding for this factor from a Xenopus liver cDNA library by using a rat LFB1 cDNA probe. A comparison of the primary structures of the Xenopus and mammalian proteins shows that the myosin-like dimerization helix, the POU-A-related domain, the homeo domain-related region, and the serine/threonine-rich activation domain are conserved between X. laevis and mammals, suggesting that all these features typical for LFB1 are essential for function. Using monoclonal antibodies, we demonstrate that XLFB1 is present not only in the liver but also in the stomach, intestine, colon, and kidney. In an analysis of the expression of XLFB1 in the developing Xenopus embryo, XLFB1 transcripts appear at the gastrula stage. The XLFB1 protein can be identified in regions of the embryo in which the liver diverticulum, stomach, gut, and pronephros are localized. The early appearance of XLFB1 expression during embryogenesis suggests that the tissue-specific transcription factor XLFB1 is involved in the determination and/or differentiation of specific cell types during organogenesis. PMID- 8417341 TI - Induction of liver alpha-1 acid glycoprotein gene expression involves both positive and negative transcription factors. AB - Expression of the alpha-1 acid glycoprotein (AGP) gene is liver specific and acute phase responsive. Within the 180-bp region of the AGP promoter, at least five cis elements have been found to interact with trans-acting factors. Four of these elements (A, C, D, and E) interacted with AGP/EBP, a liver-enriched transcription factor, as shown by footprinting analysis and by an anti-AGP/EBP antibody-induced supershift in a gel retardation assay. Modification of these sites by site-directed mutagenesis coupled with transfection analysis indicated that AGP/EBP binding to all of these sites resulted in positive regulation of the promoter. Dose-response data suggest that AGP/EBP binding to these sites results in the cooperative activation of the promoter. In contrast, functional assays showed that element B is a negative regulatory element; this element is recognized by heat-stable DNA-binding factors which are found in many cells and tissues. The regulation of these binding proteins was studied in rat liver treated with lipopolysaccharide (LPS), which induced an acute-phase reaction. We found that LPS treatment resulted in a two- to threefold increase in AGP/EBP activity and a severalfold decrease in the activity of factors that bind to element B in the liver. These results indicate that expression of the AGP gene can be regulated by both positive and negative factors and that the modulation of these factors can account for the LPS induction of the AGP gene. PMID- 8417342 TI - Discrimination among potential activators of the beta-globin CACCC element by correlation of binding and transcriptional properties. AB - Adult beta-globin-like promoters contain a cis-acting element, CCACACCC, that is conserved across species and is required for wild-type levels of transcription. We have studied the contribution of this element and proteins that interact with it to activate beta-globin transcription. We found that an erythroid-like cell line, MEL, contains several proteins that specifically bind the CACCC element. By comparing the DNA-binding properties of promoters with mutations in the CACCC element with the transcriptional activities of these mutant promoters, we found that two CACCC-binding proteins did not bind to mutant promoters that direct decreased levels of transcription. One of these proteins is the transcriptional activator Sp1, and the other we have designated CACD (CACCC-binding species D). We subjected CACD to a binding site selection procedure and obtained high affinity CACD binding sites that are identical to that of the beta-globin CACCC element. This result, combined with our finding that CACD binds the CACCC element with a higher affinity than does Sp1, argues that the CACCC element is a target of CACD rather than Sp1. The strategy of correlating the results of a binding site selection experiment with those of in vivo expression and in vitro binding studies may allow evaluation of the relative potential of different proteins to activate transcription through a single cis-acting site. PMID- 8417343 TI - Hepatocyte-specific expression of the hepatitis B virus core promoter depends on both positive and negative regulation. AB - The core promoter of hepatitis B virus shows hepatocyte specificity, which is largely dependent on an upstream regulatory sequence that overlaps with viral enhancer II. Footprint analyses by numerous groups have shown binding by cellular proteins over a large stretch of DNA in this region, but the identity of these proteins and their role in core promoter function remain largely unknown. We present data showing that the transcription factor HNF-4 is one such factor, as it activates the core promoter approximately 20-fold via a binding site within the upstream regulatory sequence. Since HNF-4 is enriched in hepatocytes, its involvement at least partially explains the hepatocyte specificity of this promoter. In addition, however, we have found a region upstream of the HNF-4 site that suppresses activation by HNF-4 in HeLa cells but not in hepatoma cells. Therefore, the cell type specificity of the core promoter appears to result from a combination of activation by one or more factors specifically enriched in hepatocytes and repression by some other factor(s) present in nonhepatocytes, and it may provide a convenient model system for studying this type of tissue specific transcriptional regulation in mammalian cells. PMID- 8417344 TI - Estradiol-inducible squelching and cell growth arrest by a chimeric VP16-estrogen receptor expressed in Saccharomyces cerevisiae: suppression by an allele of PDR1. AB - We have constructed and characterized a flexible system for analyzing the phenomenon of squelching and estrogen receptor function in the yeast Saccharomyces cerevisiae. The A/B region of the human estrogen receptor was replaced with the transcriptional activating domain of VP16 and expressed in yeast cells from high-copy-number plasmids. Addition of hormone resulted in an immediate inhibition of expression (squelching) of a chromosomally integrated GAL1:lacZ reporter gene and the eventual arrest of cell growth (toxicity). In order to determine whether a relationship exists between toxicity and squelching, mutations were made in this chimeric receptor (VEO) and their effects on transcriptional activation, squelching, and toxicity were compared. A direct correlation was found between mutations in VEO that reduced VP16 transactivation ability in yeast cells and those that reduced both squelching and toxicity. Surprisingly, mutations in the DNA binding domain (DBD) of VEO dramatically reduced squelching and completely relieved toxicity, suggesting a role for the DBD in squelching and strengthening the correlation between squelching and toxicity. To demonstrate the utility of this system for carrying out genetic selection, a plasmid-based yeast genomic bank was screened for genes that can relieve the toxicity of VEO by means of an elevated copy number, resulting in the repeated cloning of an allele of the PDR1 (pleiotropic drug resistance) gene. We present evidence that mutations in PDR1 can modulate the intracellular availability of estradiol by the same mechanism that leads to multiple drug resistance in yeast cells. Taken together, our results provide evidence that cell growth arrest occurs when squelching exceeds a certain threshold and that strong squelching requires both a DBD and a transcriptional activating domain. Furthermore, we show that growth arrest can provide a useful phenotype for carrying out the genetic analysis of both squelching and estrogen receptor function in yeast cells. PMID- 8417345 TI - Hematopoietic commitment during embryonic stem cell differentiation in culture. AB - We report that embryonic stem cells efficiently undergo differentiation in vitro to mesoderm and hematopoietic cells and that this in vitro system recapitulates days 6.5 to 7.5 of mouse hematopoietic development. Embryonic stem cells differentiated as embryoid bodies (EBs) develop erythroid precursors by day 4 of differentiation, and by day 6, more than 85% of EBs contain such cells. A comparative reverse transcriptase-mediated polymerase chain reaction profile of marker genes for primitive endoderm (collagen alpha IV) and mesoderm (Brachyury) indicates that both cell types are present in the developing EBs as well in normal embryos prior to the onset of hematopoiesis. GATA-1, GATA-3, and vav are expressed in both the EBs and embryos just prior to and/or during the early onset of hematopoiesis, indicating that they could play a role in the early stages of hematopoietic development both in vivo and in vitro. The initial stages of hematopoietic development within the EBs occur in the absence of added growth factors and are not significantly influenced by the addition of a broad spectrum of factors, including interleukin-3 (IL-3), IL-1, IL-6, IL-11, erythropoietin, and Kit ligand. At days 10 and 14 of differentiation, EB hematopoiesis is significantly enhanced by the addition of both Kit ligand and IL-11 to the cultures. Kinetic analysis indicates that hematopoietic precursors develop within the EBs in an ordered pattern. Precursors of the primitive erythroid lineage appear first, approximately 24 h before precursors of the macrophage and definitive erythroid lineages. Bipotential neutrophil/macrophage and multilineage precursors appear next, and precursors of the mast cell lineage develop last. The kinetics of precursor development, as well as the growth factor responsiveness of these early cells, is similar to that found in the yolk sac and early fetal liver, indicating that the onset of hematopoiesis within the EBs parallels that found in the embryo. PMID- 8417346 TI - A conserved alternative splice in the von Recklinghausen neurofibromatosis (NF1) gene produces two neurofibromin isoforms, both of which have GTPase-activating protein activity. AB - Sequence analysis has shown significant homology between the catalytic regions of the mammalian ras GTPase-activating protein (GAP), yeast Ira1p and Ira2p (inhibitory regulators of the RAS-cyclic AMP pathway), and neurofibromin, the protein encoded by the NF1 gene. Yeast expression experiments have confirmed that a 381-amino-acid segment of neurofibromin, dubbed the GAP-related domain (GRD), can function as a GAP. Using the RNA polymerase chain reaction with primers flanking the NF1-GRD, we have identified evidence for alternative splicing in this region of the NF1 gene. In addition to the already published sequence (type I), an alternative RNA carrying a 63-nucleotide insertion (type II) is present in all tissues examined, although the relative amounts of types I and II vary. The insertion is conserved across species but is not present in GAP, IRA1, or IRA2. GenBank searches have failed to identify significant similarity between the inserted sequence and known DNA or protein sequences, although the basic amino acid composition of the insertion shares features with nuclear targeting sequences. Expression studies in yeasts show that despite the partial disruption of the neurofibromin-IRA-GAP homology by this insertion, both forms of the NF1 GRD can complement loss of IRA function. In vivo assays designed to compare the GAP activity of the two alternatively spliced forms of the NF1-GRD show that both can increase the conversion of GTP-bound ras to its GDP-bound form, although the insertion of the 21 amino acids weakens this effect. The strong conservation of this alternative splicing suggests that both type I and II isoforms mediate important biological functions of neurofibromin. PMID- 8417347 TI - DNA polymerases delta and epsilon are required for chromosomal replication in Saccharomyces cerevisiae. AB - Three DNA polymerases, alpha, delta, and epsilon are required for viability in Saccharomyces cerevisiae. We have investigated whether DNA polymerases epsilon and delta are required for DNA replication. Two temperature-sensitive mutations in the POL2 gene, encoding DNA polymerase epsilon, have been identified by using the plasmid shuffle technique. Alkaline sucrose gradient analysis of DNA synthesis products in the mutant strains shows that no chromosomal-size DNA is formed after shift of an asynchronous culture to the nonpermissive temperature. The only DNA synthesis observed is a reduced quantity of short DNA fragments. The DNA profiles of replication intermediates from these mutants are similar to those observed with DNA synthesized in mutants deficient in DNA polymerase alpha under the same conditions. The finding that DNA replication stops upon shift to the nonpermissive temperature in both DNA polymerase alpha- and DNA polymerase epsilon- deficient strains shows that both DNA polymerases are involved in elongation. By contrast, previous studies on pol3 mutants, deficient in DNA polymerase delta, suggested that there was considerable residual DNA synthesis at the nonpermissive temperature. We have reinvestigated the nature of DNA synthesis in pol3 mutants. We find that pol3 strains are defective in the synthesis of chromosomal-size DNA at the restrictive temperature after release from a hydroxyurea block. These results demonstrate that yeast DNA polymerase delta is also required at the replication fork. PMID- 8417348 TI - GCD11, a negative regulator of GCN4 expression, encodes the gamma subunit of eIF 2 in Saccharomyces cerevisiae. AB - The eukaryotic translation initiation factor eIF-2 plays a critical role in regulating the expression of the yeast transcriptional activator GCN4. Mutations in genes encoding the alpha and beta subunits of eIF-2 alter translational efficiency at the GCN4 AUG codon and constitutively elevate GCN4 translation. Mutations in the yeast GCD11 gene have been shown to confer a similar phenotype. The nucleotide sequence of the cloned GCD11 gene predicts a 527-amino-acid polypeptide that is similar to the prokaryotic translation elongation factor EF Tu. Relative to EF-Tu, the deduced GCD11 amino acid sequence contains a 90-amino acid N-terminal extension and an internal cysteine-rich sequence that contains a potential metal-binding finger motif. We have identified the GCD11 gene product as the gamma subunit of eIF-2 by the following criteria: (i) sequence identities with mammalian eIF-2 gamma peptides; (ii) increased eIF-2 activity in extracts prepared from cells cooverexpressing GCD11, eIF-2 alpha, and eIF-2 beta; and (iii) cross-reactivity of antibodies directed against the GCD11 protein with the 58-kDa polypeptide present in purified yeast eIF-2. The predicted GCD11 polypeptide contains all of the consensus elements known to be required for guanine nucleotide binding, suggesting that, in Saccharomyces cerevisiae, the gamma subunit of eIF-2 is responsible for GDP-GTP binding. PMID- 8417349 TI - Replication and mutagenesis of UV-damaged DNA templates in human and monkey cell extracts. AB - We have used in vitro DNA replication systems from human HeLa cells and monkey CV 1 cells to replicate a UV-damaged simian virus 40-based shuttle vector plasmid, pZ189. We found that replication of the plasmid was inhibited in a UV fluence dependent manner, but even at UV fluences which caused damage to essentially all of the plasmid molecules some molecules became completely replicated. This replication was accompanied by an increase (up to 15-fold) in the frequency of mutations detected in the supF gene of the plasmid. These mutations were predominantly G:C-->A:T transitions similar to those observed in vivo. Treatment of the UV-irradiated plasmid DNA with Escherichia coli photolyase to reverse pyrimidine cyclobutane dimers (the predominant UV-induced photoproduct) before replication prevented the UV-induced inhibition of replication and reduced the frequency of mutations in supF to background levels. Therefore, the presence of pyrimidine cyclobutane dimers in the plasmid template appears to be responsible for both inhibition of replication and mutation induction. Further analysis of the replication of the UV-damaged plasmid revealed that closed circular replication products were sensitive to T4 endonuclease V (a pyrimidine cyclobutane dimer-specific endonuclease) and that this sensitivity was abolished by treatment of the replicated DNA with E. coli photolyase after replication but before T4 endonuclease treatment. These results demonstrate that these closed circular replication products contain pyrimidine cyclobutane dimers. Density labeling experiments revealed that the majority of plasmid DNA synthesized in vitro in the presence of bromodeoxyuridine triphosphate was hybrid density whether or not the plasmid was treated with UV radiation before replication; therefore, replication of UV-damaged templates appears to occur by the normal semiconservative mechanism. All of these data suggest that replication of UV damaged templates occurs in vitro as it does in vivo and that this replication results in mutation fixation. PMID- 8417350 TI - Concerted action of the transcriptional activators REB1, RAP1, and GCR1 in the high-level expression of the glycolytic gene TPI. AB - In Saccharomyces cerevisiae, the TPI gene product, triosephosphate isomerase, makes up about 2% of the soluble cellular protein. Using in vitro and in vivo footprinting techniques, we have identified four binding sites for three factors in the 5' noncoding region of TPI: a REB1-binding site located at positions -401 to -392, two GCR1-binding sites located at positions -381 to -366 and -341 to 326, and a RAP1-binding site located at positions -358 to -346. We tested the effects of mutations at each of these binding sites on the expression of a TPI::lacZ gene fusion which carried 853 bp of the TPI 5' noncoding region integrated at the URA3 locus. The REB1-binding site is dispensable when material 5' to it is deleted; however, if the sequence 5' to the REB1-binding site is from the TPI locus, expression is reduced fivefold when the site is mutated. Because REB1 blocks nucleosome formation, the most likely function of its binding site in the TPI controlling region is to prevent the formation of nucleosomes over the TPI upstream activation sequence. Mutations in the RAP1-binding site resulted in a 10-fold reduction in expression of the reporter gene. Mutating either GCR1 binding site alone had a modest effect on expression of the fusion. However, mutating both GCR1-binding sites resulted in a 68-fold reduction in the level of expression of the reporter gene. A LexA-GCR1 fusion protein containing the DNA binding domain of LexA fused to the amino terminus of GCR1 was able to activate expression of a lex operator::GAL1::lacZ reporter gene 116-fold over background levels. From this experiment, we conclude that GCR1 is able to activate gene expression in the absence of REB1 or RAP1 bound at adjacent binding sites. On the basis of these results, we suggest that GCR1 binding is required for activation of TPI and other GCR1-dependent genes and that the primary role of other factors which bind adjacent to GCR1-binding sites is to facilitate of modulate GCR1 binding in vivo. PMID- 8417351 TI - Unusually large telomeric repeats in the yeast Candida albicans. AB - We have identified sequences at the telomeres of the yeast Candida albicans and have found that they are composed of tandem copies of a 23-bp sequence. Through the cloning of native telomeric ends and the characterization and cloning of a "healed" end, we demonstrate that these repeated sequences are sufficient to function as a telomere. All copies of the 23-bp repeat that have been sequenced from a number of C. albicans strains are identical. In contrast, adjacent subtelomeric sequences are variable both between strains and within the WO-1 strain. In the WO-1 strain, the lengths of the telomeres are dependent upon growth temperature and are substantially longer at higher temperatures. Telomere growth is accompanied by increases in the number of the 23-bp repeats present on the telomeric fragments. These results suggest that either telomerase-maintained telomeres can be more complex in structure than was previously imagined or that Candida telomeres are maintained via a telomerase-independent mechanism. PMID- 8417352 TI - Transcriptional activation by the adenovirus larger E1a product is mediated by members of the cellular transcription factor ATF family which can directly associate with E1a. AB - We recently isolated three cDNA clones encoding closely related proteins (ATFa1, ATFa2, and ATFa3) that belong to the activating transcription factor-cyclic AMP responsive element family of cellular transcription factors. Using cotransfection experiments, we showed that these proteins mediate the transcriptional activation induced by the adenovirus E1a 13S mRNA gene product and that the zinc-binding domains present in both E1a conserved region 3 and the most N-terminal portion of the ATFa proteins play crucial roles in this activity. Reciprocal coimmunoprecipitation experiments demonstrated direct interactions between these proteins. Neither the conserved region 3 domain of E1a nor the N-terminal metal binding element of ATFa is essential for these interactions. The simultaneous alteration of both the N-terminal and the C-terminal domains of ATFa abolished E1a binding, while either mutation alone failed to impair these interactions. PMID- 8417353 TI - Influence of CpG methylation and target spacing on V(D)J recombination in a transgenic substrate. AB - We have previously described a line of transgenic mice with multiple head-to-tail copies of an artificial V-J recombination substrate and have shown that the methylation of this transgene is under the control of a dominant strain-specific modifier gene, Ssm-1. When the transgene array is highly methylated, no recombination is detectable, but when it is unmethylated, V-J joining is seen in the spleen, bone marrow, lymph nodes, and Peyer's patches but not in the thymus or nonlymphoid tissues, including brain tissue. Strikingly, in mice with partially methylated transgene arrays, rearrangement preferentially occurs in hypomethylated copies. Therefore, V-J recombination is negatively correlated with methylated DNA sequences. In addition, it appears that recombination occurs randomly between any two recombination signal sequences within the transgene array. This lack of target preference in an unselectable array of identical targets rules out simple mechanisms of one-dimensional tracking of a V(D)J recombinase complex. PMID- 8417354 TI - Characterization of the mouse beta maj globin transcription termination region: a spacing sequence is required between the poly(A) signal sequence and multiple downstream termination elements. AB - For the majority of mRNA encoding eukaryotic transcription units, there is little or no knowledge of the elements responsible for transcription termination or how they may interact with RNA polymerase. In this report, we have used recombinant adenovirus reporter vectors to characterize the mouse beta maj globin sequence elements that cause transcription termination. Within the globin 3' termination region, we have identified at least three sequence elements which induce significant levels of transcription termination (> 50%). The smallest functionally active element (64% termination) is 69 bp in length. The natural arrangement of these elements results in a cumulative termination which is greater than 90%. Recognition of the termination elements by RNA polymerase II depends on the presence of a functional poly(A) signal sequence. We demonstrate that efficient transcription termination depends on appropriate spacing between the poly(A) signal sequence and the termination element. PMID- 8417355 TI - fos/jun repression of cardiac-specific transcription in quiescent and growth stimulated myocytes is targeted at a tissue-specific cis element. AB - Unlike that of skeletal muscle cells in which growth and differentiation appear mutually exclusive, growth stimulation of cardiac cells is characterized by transient expression of early response nuclear proto-oncogenes as well as induction of several cardiac-specific markers. This observation led to the speculation that these proto-oncogenes, particularly c-fos and c-jun, might act as positive regulators of cardiac transcription. We have examined the role of c jun and c-fos in basal and growth-stimulated cardiac transcription, using the cardiac-specific atrial natriuretic factor (ANF) gene as a marker. The results indicate that c-jun and c-fos are negative regulators of ANF transcription. Inducers of jun and fos activity, such as mitogens and growth factors, inhibited endogenous ANF transcripts. In transient cotransfection assays, jun and fos were able to trans-repress the ANF promoter in both quiescent and alpha 1-adrenergic stimulated myocytes. This repression was specific to myocyte cultures and was not observed in nonmuscle cells. Deletion analysis indicated that repression does not require typical AP-1-binding sites (tetradecanoyl phorbol acetate response elements) or serum response elements but is targeted at a cardiac-specific element within the ANF promoter. Various Fos-related proteins, including Fra-1, Fos B, and v-Fos, were able to trans-repress ANF transcription. In addition, C terminal c-fos mutants which no longer repress transcription of such early growth response genes as c-fos and EGR-1 retained the ability to repress ANF transcription. Repression by c-jun occurs via the N-terminal activation domain and does not require the DNA-binding domain, suggesting that proto-oncogene repression involves interaction with one or more limiting cardiac-specific coactivators. PMID- 8417356 TI - Profilaggrin is a major epidermal calcium-binding protein. AB - Profilaggrin is a major highly phosphorylated protein component of the keratohyalin granules of mammalian epidermis. It contains 10 to 12 tandemly repeated filaggrin units and is processed into the intermediate filament associated protein filaggrin by specific dephosphorylation and proteolysis during terminal differentiation of the epidermal cells. Later, filaggrin itself is degraded to free amino acids that participate in maintenance of epidermal flexibility. The present paper describes the structural organization of the 5' region of the human profilaggrin gene as well as the amino terminus of the profilaggrin protein. The primary profilaggrin transcript consists of three exons and two introns. The first exon (exon I) is only 54 bp and is untranslated. The coding sequences are distributed between exon II (159 bp) and exon III, which contains the information for 10 to 12 filaggrin repeats (972 bp each) and the 3' noncoding sequences. A very large intron separates exons I and II. The combination of a very short exon I with an unusually long intron 1 makes the structure of the profilaggrin gene unique among the epidermally expressed genes investigated so far. Comparison of the expression patterns revealed by primer extension and RNase protection analysis of foreskin epidermal and cultured keratinocyte RNAs suggests that alternately spliced messages, which are different from profilaggrin mRNA, are transcribed from the profilaggrin gene system at earlier stages of epidermal differentiation. The amino terminus of profilaggrin exhibits a significant homology to the small calcium-binding S100-like proteins. It contains two alpha-helical regions, termed EF-hands, that bind calcium in vitro. This is the first example of functional calcium-binding domains fused to a structural protein. We suggest that in addition to its role in filament aggregation and the maintenance of epidermal flexibility, profilaggrin may play an important role in the differentiation of the epidermis by autoregulating its own processing in a calcium-dependent manner or by participating in the transduction of calcium signal in epidermal cells. PMID- 8417357 TI - Splicing in Caenorhabditis elegans does not require an AG at the 3' splice acceptor site. AB - The dinucleotide AG, found at the 3' end of virtually all eukaryotic pre-mRNA introns, is thought to be essential for splicing. Reduction-of-function mutations in two Caenorhabditis elegans genes, the receptor tyrosine kinase gene let-23 and the collagen gene dpy-10, both alter the AG at the end of a short (ca. 50 nucleotide) intron to AA. The in vivo effects of these mutations were studied by sequencing polymerase chain reaction-amplified reverse-transcribed RNA isolated from the two mutants. As expected, we find transcripts that splice to a cryptic AG, skip an exon, and retain an unspliced intron. However, we also find significant levels of splicing at the mutated 3' splice site (AA) and at nearby non-AG dinucleotides. Our results indicate that for short C. elegans introns an AG is not required for splicing at either the correct 3' splice site or incorrect sites. Analysis of a splice site mutant involving a longer, 316-nucleotide C. elegans intron indicates that an AG is also not required there for splicing. We hypothesize that elements besides the invariant AG, e.g., an A-U-rich region, a UUUC motif, and/or a potential branch point sequence, are directing the selection of the 3' splice site and that in wild-type genes these elements cooperate so that proper splicing occurs. PMID- 8417358 TI - Roles of multiple glucose transporters in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, TRK1 and TRK2 are required for high- and low affinity K+ transport. Among suppressors of the K+ transport defect in trk1 delta trk2 delta cells, we have identified members of the sugar transporter gene superfamily. One suppressor encodes the previously identified glucose transporter HXT1, and another encodes a new member of this family, HXT3. The inferred amino acid sequence of HXT3 is 87% identical to that of HXT1, 64% identical to that of HXT2, and 32% identical to that of SNF3. Like HXT1 and HXT2, overexpression of HXT3 in snf3 delta cells confers growth on low-glucose or raffinose media. The function of another new member of the HXT superfamily, HXT4 (previously identified by its ability to suppress the snf3 delta phenotype; L. Bisson, personal communication), was revealed in experiments that deleted all possible combinations of the five members of the glucose transporter gene family. Neither SNF3, HXT1, HXT2, HXT3, nor HXT4 is essential for viability. snf3 delta hxt1 delta hxt2 delta hxt3 delta hxt4 delta cells are unable to grow on media containing high concentrations of glucose (5%) but can grow on low-glucose (0.5%) media, revealing the presence of a sixth transporter that is itself glucose repressible. This transporter may be negatively regulated by SNF3 since expression of SNF3 abolishes growth of hxt1 delta hxt2 delta hxt3 delta hxt4 delta cells on low-glucose medium. HXT1, HXT2, HXT3, and HXT4 can function independently: expression of any one of these genes is sufficient to confer growth on medium containing at least 1% glucose. A synergistic relationship between SNF3 and each of the HXT genes is suggested by the observation that SNF2 hxt1 delta hxt2 delta hxt3 delta hxt4 delta cells and snf3 delta HXT1 HXT2 HXT3 HXT4 cells are unable to grow on raffinose (low fructose) yet SNF3 in combination with any single HXT gene is sufficient for growth on raffinose. HXT1 and HXT3 are differentially regulated. HXT1::lacZ is maximally expressed during exponential growth whereas HXT3::lacZ is maximally expressed after entry into stationary phase. PMID- 8417359 TI - The REB1 site is an essential component of a terminator for RNA polymerase I in Saccharomyces cerevisiae. AB - We have identified a terminator for transcription by RNA polymerase I in the genes coding for rRNA of the yeast Saccharomyces cerevisiae. The terminator is located 108 bp downstream of the 3' end of the mature 25S rRNA and shares several characteristics with previously studied polymerase I terminators in the vertebrates. For example, the yeast terminator is orientation dependent, is inhibited by its own sequence, and forms RNA 3' ends 17 +/- 2 bp upstream of an essential protein binding site. The recognition sequence for binding of the previously cloned REB1 protein (Q. Ju, B. E. Morrow, and J. R. Warner, Mol. Cell. Biol. 10:5226-5234, 1990) is an essential component of the terminator. In addition, the efficiency of termination depends upon sequence context extending at least 12 bp upstream of the REB1 site. PMID- 8417360 TI - GATA and Ets cis-acting sequences mediate megakaryocyte-specific expression. AB - The human glycoprotein IIB (GPIIB) gene is expressed only in megakaryocytes, and its promoter displays cell type specificity. We show that this specificity involved two cis-acting sequences. The first one, located at -55, contains a GATA binding site. Point mutations that abolish protein binding on this site decrease the activity of the GPIIB promoter but do not affect its tissue specificity. The second one, located at -40, contains an Ets consensus sequence, and we show that Ets-1 or Ets-2 protein can interact with this -40 GPIIB sequence. Point mutations that impair Ets binding decrease the activity of the GPIIB promoter to the same extent as do mutations that abolish GATA binding. A GPIIB 40-bp DNA fragment containing the GATA and Ets binding sites can confer activity to a heterologous promoter in megakaryocytic cells. This activity is independent of the GPIIB DNA fragment orientation, and mutations on each binding site result in decreased activity. Using cotransfection assays, we show that c-Ets-1 and human GATA1 can transactive the GPIIB promoter in HeLa cells and can act additively. Northern (RNA) blot analysis indicates that the ets-1 mRNA level is increased during megakaryocyte-induced differentiation of erythrocytic/megakaryocytic cell lines. Gel retardation assays show that the same GATA-Ets association is found in the human GPIIB enhancer and the rat platelet factor 4 promoter, the other two characterized regulatory regions of megakaryocyte-specific genes. These results indicate that GATA and Ets cis-acting sequences are an important determinant of megakaryocytic specific gene expression. PMID- 8417361 TI - Cell cycle analysis of p53-induced cell death in murine erythroleukemia cells. AB - A temperature-sensitive mutant of murine p53 (p53Val-135) was transfected by electroporation into murine erythroleukemia cells (DP16-1) lacking endogenous expression of p53. While the transfected cells grew normally in the presence of mutant p53 (37.5 degrees C), wild-type p53 (32.5 degrees C) was associated with a rapid loss of cell viability. Genomic DNA extracted at 32.5 degrees C was seen to be fragmented into a characteristic ladder consistent with cell death due to apoptosis. Following synchronization by density arrest, transfected cells released into G1 at 32.5 degrees C were found to lose viability more rapidly than did randomly growing cultures. Following release into G1, cells became irreversibly committed to cell death after 4 h at 32.5 degrees C. Commitment to cell death correlated with the first appearance of fragmented DNA. Synchronized cells allowed to pass out of G1 prior to being placed at 32.5 degrees C continued to cycle until subsequently arrested in G1; loss of viability occurred following G1 arrest. In contrast to cells in G1, cells cultured at 32.5 degrees C for prolonged periods during S phase and G2/M, and then returned to 37.5 degrees C, did not become committed to cell death. G1 arrest at 37.5 degrees C, utilizing either mimosine or isoleucine deprivation, does not lead to rapid cell death. Upon transfer to 32.5 degrees C, these G1 synchronized cell populations quickly lost viability. Cells that were kept density arrested at 32.5 degrees C (G0) lost viability at a much slower rate than did cells released into G1. Taken together, these results indicate that wild-type p53 induces cell death in murine erythroleukemia cells and that this effect occurs predominantly in the G1 phase of actively cycling cells. PMID- 8417362 TI - Involvement of rho p21 and its inhibitory GDP/GTP exchange protein (rho GDI) in cell motility. AB - Evidence is accumulating that rho p21, a ras p21-related small GTP-binding protein (G protein), regulates the actomyosin system. The actomyosin system is known to be essential for cell motility. In the present study, we examined the action of rho p21, its inhibitory GDP/GTP exchange protein (named rho GDI), its stimulatory GDP/GTP exchange protein (named smg GDS), and Clostridium botulinum ADP-ribosyltransferase C3, known to selectively ADP-ribosylate rho p21 and to impair its function, in cell motility (chemokinesis) of Swiss 3T3 cells. We quantitated the capacity of cell motility by measuring cell tracks by phagokinesis. Microinjection of the GTP gamma S-bound active form of rhoA p21 or smg GDS into Swiss 3T3 cells did not affect cell motility, but microinjection of rho GDI into the cells did inhibit cell motility. This rho GDI action was prevented by comicroinjection of rho GDI with the GTP gamma S-bound form of rhoA p21 but not with the same form of rhoA p21 lacking the C-terminal three amino acids which was not posttranslationally modified with lipids. The rho GDI action was not prevented by Ki-rasVal-12 p21 or any of the GTP gamma S-bound form of other small GTP-binding proteins including rac1 p21, G25K, and smg p21B. Among these small G proteins, rhoA p21, rac1 p21, and G25K are known to be substrates for rho GDI. The rho GDI action was not prevented by comicroinjection of rho GDI with smg GDS. Microinjection of C3 into Swiss 3T3 cells also inhibited cell motility. These results indicate that the rho GDI-rho p21 system regulates cell motility, presumably through the actomyosin system. PMID- 8417363 TI - Temporal order of RNA-processing reactions in trypanosomes: rapid trans splicing precedes polyadenylation of newly synthesized tubulin transcripts. AB - Many trypanosome genes are expressed as part of large polycistronic transcription units. This finding suggests that regulation of mRNA biogenesis may emphasize RNA processing reactions more so than in other organisms. This study was undertaken to understand the temporal order of two RNA-processing reactions, trans splicing and polyadenylation, in the maturation of trypanosome mRNAs in vivo. Kinetic studies revealed rapid trans splicing of alpha-tubulin, beta-tubulin, and actin pre-mRNAs within 1 to 2 min after synthesis of the 3' splice site. Furthermore, following blockage of pre-mRNA synthesis, newly synthesized spliced leader RNA cannot be used for trans splicing, suggesting that trypanosomes do not accumulate substantial amounts of pre-mRNA which can provide splice acceptor sites. Thus, trans splicing is cotranscriptional. In addition, we show that trans splicing precedes polyadenylation in the processing of trypanosome tubulin pre-mRNAs. PMID- 8417364 TI - Identification of a negative element in the human vimentin promoter: modulation by the human T-cell leukemia virus type I Tax protein. AB - The vimentin gene is a member of the intermediate filament multigene family and encodes a protein expressed, in vivo, in all mesenchymal derivatives and, in vitro, in cell types of various origin. We have previously demonstrated that the expression of this growth-regulated gene could be trans activated by the 40-kDa Tax protein of HTLV-I (human T-cell leukemia virus type I) and that responsiveness to this viral protein was mediated by the presence of an NF-kappa B binding site located between -241 and -210 bp upstream of the mRNA cap site (A. Lilienbaum, M. Duc Dodon, C. Alexandre, L. Gazzolo, and D. Paulin, J. Virol. 64:256-263, 1990). These previous assays, performed with deletion mutants of the vimentin promoter linked to the chloramphenicol acetyltransferase gene, also revealed the presence of an upstream negative region between -529 and -241 bp. Interestingly, the inhibitory activity exerted by this negative region was overcome after cotransfection of a Tax-expressing plasmid. In this study, we further characterize the vimentin negative element and define the effect of the Tax protein on the inhibitory activity of this element. We first demonstrate that a 187-bp domain (-424 to -237 bp) behaves as a negative region when placed upstream either of the NF-kappa B binding site of vimentin or of a heterologous enhancer such as that present in the desmin gene promoter. The negative effect can be further assigned to a 32-bp element which is indeed shown to repress the basal or induced activity of the NF-kappa B binding site.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417365 TI - An opportunistic promoter sharing regulatory sequences with either a muscle specific or a ubiquitous promoter in the human aldolase A gene. AB - The human aldolase A gene is transcribed from three different promoters, pN, pM, and pH, all of which are clustered within a small 1.6-kbp DNA domain. pM, which is highly specific to adult skeletal muscle, lies in between pN and pH, which are ubiquitous but particularly active in heart and skeletal muscle. A ubiquitous enhancer, located just upstream of pH start sites, is necessary for the activity of both pH and pN in transient transfection assays. Using transgenic mice, we studied the sequence controlling the muscle-specific promoter pM and the relations between the three promoters and the ubiquitous enhancer. A 4.3-kbp fragment containing the three promoters and the ubiquitous enhancer showed an expression pattern consistent with that known in humans. In addition, while pH was active in both fast and slow skeletal muscles, pM was active only in fast muscle. pM activity was unaltered by the deletion of a 1.8-kbp region containing the ubiquitous enhancer and the pH promoter, whereas pN remained active only in fast skeletal muscle. These findings suggest that in fast skeletal muscle, a tissue-specific enhancer was acting on both pN and pM, whereas in other tissues, the ubiquitous enhancer was necessary for pN activity. Finally, a 2.6-kbp region containing the ubiquitous enhancer and only the pH promoter was sufficient to bring about high-level expression of pH in cardiac and skeletal muscle. Thus, while pH and pM function independently of each other, pN, remarkably, shares regulatory elements with each of them, depending on the tissue. Importantly, expression of the transgenes was independent of the integration site, as originally described for transgenes containing the beta-globin locus control region. PMID- 8417366 TI - Transcriptional inhibition of the murine erythropoietin receptor gene by an upstream repetitive element. AB - Transcription of the murine erythropoietin receptor (EpoR) gene is inhibited by a novel repetitive element that is located upstream of the EpoR promoter. Reporter gene studies reveal that the inhibitory effect is both distance and orientation dependent. This element is a member of a family of repetitive elements specific to rodents and is present at approximately 10(5) copies per mouse genome. It encodes approximately 500- to 900-bp-long transcripts in both erythroid and nonerythroid cells. RNase protection analysis with a probe from the 5' flanking murine EpoR gene reveals that the direction of transcription is in the sense orientation, relative to the downstream EpoR gene. We suggest that transcriptional inhibition of the EpoR promoter is mediated by read-through transcripts originating in the upstream repetitive element and that this effect may contribute to the basal level of transcription of the murine EpoR gene in erythroid cells. PMID- 8417367 TI - Particle counting by flow cytometry can determine intrinsic equilibrium constants for antibody-ligand interactions in solution. PMID- 8417368 TI - A single amino acid substitution abolishes the heterogeneity of chimeric mouse/human (IgG4) antibody. AB - Human immunoglobulin G4 (IgG4) exists in two molecular forms due to the heterogeneity of the inter-heavy chain disulphide bridges in the hinge region in a proportion of secreted human IgG4. This heterogeneity is only revealed under denaturing, non-reducing conditions in which an HL "half antibody" is detected, a phenomenon not seen in other human IgG isotypes. In native conditions noncovalent interactions hold the antibody together as the H2L2 tetramer. Analysis of the hinge sequences of human IgG heavy chains suggested that the presence of serine at residue 241 might be the cause of this heterogeneity. We therefore changed the serine at 241 to proline (found at that position in IgG1 and IgG2) in a mouse/human chimeric heavy chain. This single residue substitution leads to the production of a homogeneous antibody. Further, the variant IgG4 has significantly extended serum half-life and shows an improved tissue distribution compared to the original chimeric IgG4. PMID- 8417369 TI - Genomic sequence of a V(D)J rearrangement utilizing a new VH7183 family member VH7183.15. PMID- 8417370 TI - Comparison of complete nucleotide sequence of the human IgM heavy chain constant region of polyreactive and monoreactive antibodies. PMID- 8417371 TI - Unstable inter-H chain disulfide bonding and non-covalently associated J chain in rat dimeric IgA. AB - Disulfide bonds are a major force in stabilizing the three-dimensional structure of immunoglobulins. To determine the pattern of interchain disulfide bonding between the four H chains, four L chains and single J chain of rat dimeric IgA (dIgA), we analyzed dIgA from the LO DNP-64 hybridoma by diagonal SDS-PAGE. Bands corresponding to one, two, three and four H chains, one and two L chains and the free J chain were observed under non-reducing conditions, suggesting that the interchain disulfide bonds in rat dIgA are unstable under denaturing conditions. Similar patterns of disulfide bonding were observed in three other hybridoma or myeloma dIgAs from LOU/CN rats. In contrast, when dIgA pretreated with iodoacetamide (IA) was analyzed by the same technique, only bands corresponding to four H chains, one and two L chains and the free J chain were observed, suggesting that blocking free sulfhydryl groups stabilizes the inter-H chain disulfide bonds. Reaction of dimeric LO DNP-64 dIgA with 5,5'-dithiobis-(2 nitrobenzoic acid) or with 14C-IA demonstrated that this dIgA contains an average of 4 moles of free sulfhydryl groups per mole of protein under non-denaturing conditions and 9 moles of free sulfhydryl groups under denaturing conditions. Taken together, the results suggest that interchain disulfide bonds in rat dIgA are unstable, presumably due to the influence of nearby free sulfhydryl groups, and that non-covalent forces are critical for stabilizing the dIgA complex. The results also indicate that J chain is entirely non-covalently associated with the H chains, an apparently unique feature of rat dIgA. A model for interchain disulfide bonding in rat dIgA is proposed. PMID- 8417372 TI - Chromosomal location and isoform analysis of mouse Fc epsilon RII/CD23. AB - The gene for the mouse low affinity receptor for IgE (Fc epsilon RII, also known as CD23) was mapped on Chromosome (Chr) 8 proximal to Plat. This gene, symbolized Fcer2 (formerly Fce2) resides in a region of Chr 8 with linkage homology with human chromosomes 8 and 19. The mouse Fc epsilon RII was examined for the presence of alternate N-terminal forms such as seen in humans. An antisense RNA probe was prepared from the 5' end of the cDNA through the first 660 bp of the cDNA and was used to analyze message from Fc epsilon RII+ B cells and B cell hybridomas both before and after treatment with interleukin 4 (IL-4). Using RNase protection analysis, a major 640 bp band corresponding to the full length probe was seen, even after activation of the cells with LPS in the presence of IL-4, which is known to give high expression levels of the Fc epsilon RII. This result suggests that the mouse does not produce significant levels of an alternate IL-4 inducible Fc epsilon RII, as seen in man, and this may explain the more restricted cell lineage expression of the Fc epsilon RII in the mouse. PMID- 8417373 TI - Human IgG3 is decreased and IgG1, IgG2 and IgG4 are unchanged in molecular size by mild reduction and reoxidation without any major change in effector functions. AB - Purified proteins of the four human IgG subclasses were reduced under neutral conditions to break the interchain S-S bonds, followed by dialysis to allow reformation of S-S bonds (pr/o treatment). The IgG1, IgG2 and IgG4 proteins apparently reformed native molecules by pr/o treatment, while IgG3 formed molecules with significantly smaller size, as measured by HPLC gel filtration, compared to the autologous native proteins. The degree of shrinking of the pr/o IgG3 molecules varied and was most pronounced at low protein concn. In addition, the temp and the concn of reducing agent during the pr/o treatment had some influence on the molecular size. The effect is probably due to a conformational change of the 62 amino acid long hinge of IgG3. The effector activity of pr/o IgG2 and pr/o IgG3 was studied by employing chimeric, mouse V and human C regions, monoclonal antibodies with the same NIP-binding properties. Thus, the interaction between IgG and the complement system was unchanged both for pr/o IgG2 and pr/o IgG3, while the Fc-receptor-mediated antibody-dependent cellular cytotoxicity (ADCC) was depressed to the same degree for both pr/o IgG2 and pr/o IgG3. Conclusively, the alteration of the conformation of the IgG3 molecule by pr/o treatment had no major influence on its effector functions. PMID- 8417374 TI - Quantitative analysis of the interaction between lysozyme and monoclonal antibody D1.3. AB - The method of sedimentation equilibrium has been used to determine the stoichiometry and binding constant for the interaction between hen egg white lysozyme and monoclonal antibody D1.3. The procedures described allow the relative binding affinities of 125I-labelled lysozyme and unlabelled lysozyme to be compared. The data indicate that labelled and unlabelled lysozyme bind to monoclonal D1.3 with similar affinity (binding constant, K = 1.6 x 10(9)/M). Using solid-phase methods estimates obtained for the binding constant were lower and dependent both on the amount of antigen coated on the plate and the dilution of primary antibody (D1.3). These data were not consistent with a simple equilibrium binding model, suggesting kinetic or orientation effects. In contrast sedimentation equilibrium experiments provide a rapid and accurate method for determining both the stoichiometry and binding constants for the interaction between antigens and antibodies. PMID- 8417375 TI - A soluble form of the human CD8 alpha chain expressed in the baculovirus system: biochemical characterization and binding to MHC class I. AB - We have generated a soluble form of the CD8 molecule consisting of the entire extracellular domains of the human alpha chain, by expressing a mutated CD8 alpha cDNA in SF9 cells infected with a recombinant baculovirus. The truncated molecule was secreted into the medium mostly as a disulfide-linked homodimer in which a single cysteine residue in the hinge-like region (Cys143) was sufficient to assure covalent bonding. Soluble CD8 purified to homogeneity appears to be monodisperse as assessed by gel filtration analysis and contains only O-linked carbohydrates. To determine whether recombinant CD8 can interact with MHC class I molecules, we developed an assay that measures binding of MHC class I-bearing cell lines to purified CD8 adsorbed to plastic plates. The level of binding of cells to immobilized CD8 depended on the amount of CD8 bound to the plate and correlated with the levels of cell surface MHC class I expression. The binding was specifically inhibited by monoclonal antibodies directed either against CD8 or MHC class I molecules. This assay therefore provides a way to measure CD8 binding to MHC class I independently of other cell-cell interactions and should allow direct structure-function studies. PMID- 8417376 TI - HLA-DQB1 typing of north east Italian IDDM patients using amplified DNA, oligonucleotide probes and a rapid DNA-enzyme immunoassay (DEIA). AB - We report on HLA-DQB1 typing in IDDM patients of north east Italian region using an enzymatic method based on the detection of hybridization reaction between PCR amplified DNA from whole blood and allele specific oligonucleotides by an antibody directed against double stranded DNA (DNA-enzyme immunoassay or DEIA). The method is reliable, simple and sensitive as the classical radioactive method with the advantage of using a universal non radioactive detection reagent. Nineteen families, each including one subject with juvenile insulin-dependent diabetes mellitus (IDDM) were analyzed. A strong association between absence of an aspartic acid (Asp) in position 57 of DQB1 beta chain in homozygous conditions and susceptibility to IDDM was found. In contrast with some previous observations, however, no significant association was found between Asp/non-Asp heterozygous genotype and IDDM. No patients were found with an homozygous Asp/Asp genotype, known to be protective in caucasoid population. Of particular interest was the DQB1 allelic distribution in our population sample. The non-Asp allele most frequently found in IDDM subjects was the DQB1 0201 allele and this finding was statistically significant (Pc value < 0.05, relative risk = 5.01). No significant association was found for any other allele including the DQB1 0302 (Pc value = not significant although with relative risk = 3.28) previously reported as the most frequent allele in IDDM caucasoid patients. PMID- 8417377 TI - Molecular and biochemical analyses of combining sites of monoclonal anti-morphine antibodies. AB - V region nucleotide sequences were determined by mRNA sequencing for 11 monoclonal anti-morphine antibodies with slightly different specificities for morphine-related opiates. The VH region nucleotide sequences of the antibodies MOR8, MOR33, MOR35, MOR44, MOR83, and MOR76 were classified into the VH-5 (7183) family, while the antibodies MOR39, MOR115, MOR131, MOR158 and MOR180 used VH-1 (J558) family genes. MOR39, MOR115 and MOR131 used the V lambda-1 gene for their L chain V region. MOR158 and MOR180 used the Vk-10 gene. MOR8, MOR33, OR35, MOR44, MOR76 and MOR83 used VK-21D. The antibody sets MOR158 and MOR180; MOR39 and MOR131; and MOR8, MOR33, MOR35, MOR44, MOR76 and MOR83 appeared to be somatic mutants derived from the same clones since they showed the same VH/VL usage and V(D)J recombination pattern. The pH-reactivity profiles for these antibodies revealed that the binding of morphine to the antibodies is highly dependent on the pH value of the assay solution, suggesting the importance of the electrostatic interaction between the positive charge of morphine and the negative charges at or near the combining sites. Direct UV-photoaffinity labeling with 3H-morphine was carried out in order to estimate the orientation of morphine in the combining sites. The H chains were preferentially labeled in MOR8, MOR33, MOR35, MOR76, and MOR83, whereas most of the crosslinked hapten was found in the L chains in MOR39, MOR115, MOR131, MOR158 and MOR180. Thus, these 11 antibodies were classified into two types in terms of reactivity in the photoaffinity labeling. PMID- 8417378 TI - Anti-peptide reagent identifies a primary-structure-dependent, cross-reactive idiotype expressed on heavy and light chains from a murine monoclonal anti-CD4. AB - A synthetic peptide corresponding to the second complementarity determining region (CDR2) of the immunoglobulin (Ig) variable (V) region heavy (H) chain (CDR2VH) of anti-Leu3a, a murine monoclonal antibody specific for the human CD4 molecule, was used to elicit the production of specific rabbit anti-peptide antibodies. The rabbit anti-peptide antiserum was tested for reactivity against the immunizing peptide, anti-Leu3a, and a panel of mouse monoclonal anti-CD4. Only the immunizing peptide and anti-Leu3a were recognized by ELISA, whereas the H chains of anti-Leu3a and five other monoclonal anti-CD4 preparations were recognized by Western blot analysis. These data suggest that linear structures corresponding to the CDR2VH are not normally exposed on the surface of these monoclonal antibodies and become accessible only upon unfolding of the Ig molecule. In addition, Western blot analysis demonstrated that the anti-CDR2VH peptide antiserum was able to recognize the Ig light (L) chain of anti-Leu3a. This reactivity to both H and L chains from anti-Leu3a was ascribed to a homologous five amino acid sequence region shared by the two chains. The region of homology was associated with the third framework (FR3) of the L chain and was included as a portion of the sequence in the CDR2VH synthetic peptide. This observation was confirmed by the ability of the CDR2VH anti-peptide antiserum to bind the L chains of three mouse myeloma proteins that exhibited the five amino acid sequence region of homology within their respective FR3. Together, these data provide information on the structural basis of idiotypes shared by the H and L chains from the same antibody molecule and indicate that five amino acids might be sufficient to define a minimal continuous idiotypic determinant. PMID- 8417379 TI - Mutations in the putative lipid-interaction domain of complement C9 result in defective secretion of the functional protein. AB - Complement protein C9 assembles with C5, C6, C7, C8 on the surface of target cells to form the lytic membrane attack complex (MAC). During MAC assembly and insertion into the target membrane, the hydrophilic, globular C9 partially unfolds to expose a hydrophobic lipid interaction domain. Several copies of amphiphilic C9 subsequently polymerize to form the characteristic ring-like MAC. Using a combined photoaffinity label and computer modeling approach, two amphipathic helices in a segment encompassing the amino acids 293-334 have been predicted to interact with membrane lipids. To elucidate the mechanism of C9 lipid binding and insertion, site-directed mutagenesis was used to change the amphipathic character of the helices. While some conservative amino acid replacements such as Thr307 by a Leu were tolerated and yielded fully active C9 when expressed in COS cells, successive changes of Leu305 into Val, Ala, and Glu on the hydrophobic site of the first helix gave rise to only partly or not secreted C9. All non-conservative amino acid replacements introduced on either side of the helices resulted in non-secreted C9 that was subsequently degraded intracellularly, indicating the importance of the correct folding of the presumptive transmembrane domain during biosynthesis. A natural secretion incompetent mutant was found in which Val293, located in the proposed lipid binding region, was lacking. Taken together, these findings suggest that the high incidence of homozygous C9 deficiencies may be due to a blockage in intracellular transport and secretion due to point mutations in this 'hot spot' region of the molecule. PMID- 8417380 TI - A population-based study of dementia in 85-year-olds. AB - BACKGROUND: The aim of this study was to investigate the causes, severity, and prevalence of dementia in a representative sample of 494 85-year-olds living in Gothenburg, Sweden. METHODS: The study included a psychiatric interview, neuropsychological and physical examinations, comprehensive laboratory tests, electrocardiography, chest radiography, computed tomography (CT) of the head, and analysis of cerebrospinal fluid. A person close to each subject was also interviewed. Dementia was defined according to the criteria proposed in the Diagnostic and Statistical Manual of Mental Disorders (third edition, revised), Alzheimer's disease according to the criteria of the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association, and vascular dementia according to recently proposed criteria that incorporate information from CT scanning and the patient's neurologic history. RESULTS: The prevalence of dementia was 29.8 percent (147 subjects). The condition was mild in 8.3 percent, moderate in 10.3 percent, and severe in 11.1 percent. There were no significant sex-related differences in prevalence or severity. Of the subjects with dementia, 43.5 percent had Alzheimer's disease, 46.9 percent had vascular dementia (multi-infarct dementia in 34.6 percent, dementia related to cerebral hypoperfusion in 4.1 percent, and mixed dementia in 8.2 percent), and 9.5 percent had dementia due to other causes. The three-year mortality rate was 23.1 percent in the subjects without dementia, 42.2 percent in the patients with Alzheimer's disease, and 66.7 percent in the patients with vascular dementia. Infarcts detected by CT scanning were significantly more common in the subjects with dementia than in those without it (27.9 percent vs. 12.6 percent). CONCLUSIONS: Dementia was present in nearly a third of unselected 85-year-olds in Sweden. Almost half these subjects appeared to have vascular dementia, which may currently be more amenable to prevention or treatment than Alzheimer's disease. PMID- 8417381 TI - Influence of cigarette smoking on the efficacy of radiation therapy in head and neck cancer. AB - BACKGROUND: Smoking is a risk factor for several cancers and may also limit the efficacy of treatment. In this study, we evaluated the influence of cigarette smoking during radiation therapy on the efficacy of treatment in patients with head and neck cancer. METHODS: Using a questionnaire, we obtained information on smoking behavior at base line and weekly during therapy in 115 patients with head and neck cancer who were treated with radiation therapy with or without fluorouracil. The side effects of therapy were evaluated weekly, and response was assessed 13 weeks after treatment was completed. The main outcomes measured were treatment response and survival. RESULTS: The prognostic variables were similar among the patients who smoked and those who did not smoke during treatment. The 53 patients who continued to smoke during radiation therapy had a lower rate of complete response (45 percent vs. 74 percent, P = 0.008) and poorer two-year survival (39 percent vs. 66 percent, P = 0.005) than the 62 patients who did not smoke or who had quit before treatment. Among the nonsmoking patients, mortality was influenced by the length of time between quitting and treatment, with a risk reduction (relative to that for patients who continued to smoke) of 40 percent for patients who had quit less than 12 weeks before diagnosis and of 70 percent for patients who had quit more than 1 year before diagnosis. After adjustment for other variables with proportional-hazards regression analysis, smoking remained an independent prognostic factor (P = 0.002), with a relative risk of 2.5 (95 percent confidence interval, 1.4 to 4.4) favoring the patients who abstained from smoking. The results could not be explained by the type of chemotherapy received, the presence of coexisting morbid conditions, differences in the side effects of radiation, or the number of interruptions of treatment. CONCLUSIONS: Patients with head and neck cancer who continue to smoke during radiation therapy have lower rates of response and survival than patients who do not smoke during radiation therapy. PMID- 8417382 TI - A preliminary study of diltiazem in the prevention of coronary artery disease in heart-transplant recipients. AB - BACKGROUND: Accelerated coronary artery disease is a major cause of late morbidity and mortality among heart-transplant recipients. Because calcium channel blockers can suppress diet-induced atherosclerosis in laboratory animals, we assessed the efficacy of diltiazem in preventing coronary artery disease in transplanted hearts. METHODS: Consecutive eligible cardiac-transplant recipients were randomly assigned to receive diltiazem (n = 52) or no calcium-channel blocker (n = 54). Coronary angiograms obtained early after cardiac transplantation and annually thereafter were used for the visual assessment of the extent of coronary artery disease. The average diameters of identical coronary artery segments were measured on the angiograms obtained at base line and at the first and second follow-up examinations. RESULTS: In the 57 patients who had all three angiograms, the average coronary artery diameter (+/- SD) 0.27 decreased in the group that received no calcium-channel blocker from 2.41 +/- 0.27 mm at base line to 2.19 +/- 0.28 mm at one year, and to 2.22 +/- 0.26 mm at two years (P < 0.001 for both years). The average diameter in the diltiazem group changed little from the base-line value of 2.32 +/- 0.22 mm (2.32 +/- 0.27 mm at one year and 2.36 +/- 0.22 mm at two years). The average change in the diameter of the segment differed significantly between the two treatment groups (P < 0.001), and the estimated effect of treatment changed only negligibly after adjustment for other relevant clinical variables. New angiographic evidence of coronary artery disease developed in 14 patients not given calcium-channel blockers, as compared with 5 diltiazem-treated patients (P = 0.082). Coronary stenoses greater than 50 percent of the luminal diameter developed in seven patients not given calcium-channel blockers, as compared with two patients given diltiazem; death due to coronary artery disease or retransplantation occurred in five patients in the group that did not receive calcium-channel blockers and none of those who received diltiazem. CONCLUSIONS: Our preliminary results suggest that diltiazem can prevent the usual reduction in the diameter of the coronary artery in cardiac-transplant recipients, but further follow-up will be required to determine whether diltiazem can decrease the long-term incidence of symptomatic coronary artery disease. PMID- 8417383 TI - Effect of serum parathyroid hormone and bone marrow fibrosis on the response to erythropoietin in uremia. AB - BACKGROUND AND METHODS: Anemia is common in patients with chronic renal insufficiency and secondary hyperparathyroidism. Erythropoietin therapy is effective, but the dose required varies greatly. One possible determinant of the efficacy of erythropoietin therapy is the extent of marrow fibrosis caused by hyperparathyroidism. We examined the relation between the erythropoietic response to erythropoietin and hyperparathyroidism in a cross-sectional study of 18 patients undergoing hemodialysis who had received erythropoietin therapy for one to three years. In 7 patients (the poor-response group), the dose of intravenous erythropoietin needed to maintain a mean (+/- SD) target hematocrit of 35 +/- 3 percent was > 100 units per kilogram of body weight three times a week, and in 11 patients (the good-response group) it was < or = 100 units per kilogram. In all patients, indexes of the adequacy of dialysis and the extent of hyperparathyroidism and aluminum toxicity were determined monthly, and bone histomorphometry was performed. RESULTS: The mean (+/- SD) dose of erythropoietin required to maintain the target hematocrit was 174 +/- 33 units per kilogram three times a week in the poor-response group and 56 +/- 18 units per kilogram in the good-response group. The mean ages, duration and adequacy of dialysis, increment in hematocrit, iron requirements, and serum concentrations of calcium, phosphate, and aluminum were similar in the two groups. The percentages of osteoid volume and surface, the osteoid thickness, and the stainable aluminum content of bone were similar in the two groups. In contrast, the mean serum parathyroid hormone concentration, the percentages of osteoclastic and eroded bone surfaces, and the degree of marrow fibrosis were greater in the poor response group than in the good-response group (P = 0.03, P = 0.04, P = 0.009, and P = 0.009, respectively). CONCLUSIONS: In patients with uremia, the dose of erythropoietin needed to achieve an adequate hematocrit response may depend on the severity of secondary hyperparathyroidism and the extent of bone marrow fibrosis. PMID- 8417384 TI - Effects of tocopherol and deprenyl on the progression of disability in early Parkinson's disease. AB - BACKGROUND AND METHODS: In 1987 we began a multicenter controlled clinical trial of deprenyl (a monoamine oxidase inhibitor) and tocopherol (a component of vitamin E that traps free radicals) in the treatment of early Parkinson's disease. We randomly assigned 800 patients to one of four treatments: placebo, active tocopherol and deprenyl placebo, active deprenyl and tocopherol placebo, or both active drugs. The primary end point was the onset of disability prompting the clinical decision to begin administering levodopa. An interim analysis showed that deprenyl was beneficial (N Engl J Med 1989;321:1364-71). We report the results of tocopherol treatment after a mean (+/- SD) follow-up of 14 +/- 6 months, as well as the follow-up results for deprenyl. RESULTS: There was no beneficial effect of tocopherol or any interaction between tocopherol and deprenyl. The beneficial effects of deprenyl, which occurred largely during the first 12 months of treatment, remained strong and significantly delayed the onset of disability requiring levodopa therapy (hazard ratio, 0.50; 95 percent confidence interval, 0.41 to 0.62; P < 0.001). The difference in the estimated median time to the end point was about nine months. The ratings for Parkinson's disease improved during the first three months of deprenyl treatment; the motor performance of deprenyl-treated patients worsened after the treatments were withdrawn. CONCLUSIONS: Deprenyl (10 mg per day) but not tocopherol (2000 IU per day) delays the onset of disability associated with early, otherwise untreated Parkinson's disease. The action of deprenyl that accounts for its beneficial effects remains unclear. PMID- 8417385 TI - Head and neck cancer. PMID- 8417386 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 3-1993. A 51-year-old Ethiopian woman with myalgia, weight loss, and mediastinal lymphadenopathy. PMID- 8417387 TI - Illnesses causing dementia in the very elderly. PMID- 8417388 TI - Investigating erythropoietin resistance. PMID- 8417389 TI - Valproate versus carbamazepine for seizures. PMID- 8417390 TI - Valproate versus carbamazepine for seizures. PMID- 8417391 TI - Valproate versus carbamazepine for seizures. PMID- 8417392 TI - Enterococcus faecalis pneumonia complicating topical antimicrobial prophylaxis. PMID- 8417393 TI - Sclerotherapy for the treatment of nodular intraoral Kaposi's sarcoma in patients with AIDS. PMID- 8417394 TI - Office-based test systems for HIV antibody. PMID- 8417395 TI - Survival of nationally shared, HLA-matched kidney transplants. PMID- 8417396 TI - The heart in hypertension. PMID- 8417397 TI - Predisposing factors for severe, uncontrolled hypertension in an inner-city minority population. PMID- 8417398 TI - Genetic factors that predispose the child to develop hypertension. AB - Familial aggregation, population, and twin studies all point to important genetic influences on the level of blood pressure in childhood and adolescence. Whether a major gene effect operates during childhood has not been determined. The investigation of polygenic paths leads to the study of variables such as ion transport and reactivity paths that appear to be under strong genetic influences. The evidence suggests that abnormalities in these paths might be linked to a prehypertensive state. Univariate genetic analyses of systolic and diastolic blood pressure show that a significant portion of the variability of these variables is under genetic control. Moreover, during early adolescence, boys differ from girls in the regulation of their resting blood pressure. Multivariate genetic analyses show that in these young adolescents, genetic paths shared with body mass index appear to influence systolic but not diastolic blood pressure. The genetic relationship between systolic and diastolic blood pressure appears largely to be independent of body mass index. Genetic studies can partition the genetic and environmental influences on blood pressure and identify shared paths with variables previously believed to be linked epidemiologically. This information may have the capacity to be the framework for public health guidelines developed to lower the incidence of adult hypertension. PMID- 8417399 TI - Hypertension in infancy. AB - Hypertension in infants is rare and requires a thorough evaluation. The incidence of hypertension in infancy has risen in recent years, reflecting both better monitoring methods and increasingly successful salvage of smaller and smaller newborns. Overall mortality and morbidity rates for uncontrolled hypertension in infants are unknown. With appropriate treatment, the prognosis for resolution of hypertension is good. In most cases, hypertension is short-lived and blood pressures return to normal even when medication is discontinued. Recent experience with improved antihypertensive agents in infancy has meant that nephrectomy for renovascular hypertension is rarely required. There is still much to learn about the indications for treatment of elevated blood pressures in infancy and the potential adverse effects of therapy. Infants with a history of neonatal hypertension should be followed closely because the long-term prognosis is not known and recurrence of hypertension remains a possibility. Because hypertension can develop in high-risk newborns following discharge from the nursery, these infants deserve routine blood pressure measurements as part of their outpatient follow-up. PMID- 8417400 TI - Renovascular hypertension in children. AB - Renal arteriography remains the only study to identify and define correctly both location and extent of renal artery stenosis. Because other studies do not adequately identify all children who have renal artery disease, it is reasonable to perform renal arteriography in those children with severe hypertension who do not have another readily identifiable cause for their hypertension. On the other hand, in children with less profound elevation of blood pressure in whom there is no other definable cause, it is possible to defer renal angiography. In these children with mild or moderate hypertension, studies such as the captopril 99mTc DTPA scan or intravenous digital subtraction angiography may identify those for whom arteriography is more likely to have abnormal results. PMID- 8417401 TI - Racial aspects of blood pressure in children and adolescents. AB - The incidence, prevalence, and severity of essential hypertension are higher in minority adult populations, especially black Americans. Studies have not uniformly shown that black children and adolescents have higher blood pressure values than whites of the same age. The goal of this article is to review data available comparing minority and white populations. A section discussing studies addressing racial differences in related topics is included to demonstrate the existing fields of research that may not be familiar to the general pediatrician. PMID- 8417402 TI - Cardiovascular causes of systemic hypertension. AB - Although cardiovascular disease is an uncommon cause of hypertension, the number one cardiovascular cause of hypertension, coarctation of the aorta, is easy to diagnose and treat. It is therefore mandatory that every individual who is evaluated for hypertension should have blood pressures measured in the arms and legs. The other, more uncommon cardiovascular causes of hypertension (Table 1), also can usually be diagnosed from a thorough history and physical examination. PMID- 8417403 TI - Endocrine causes of hypertension in children. AB - Hypertension in children is not a common problem. When it is found, however, a pathologic cause can often be identified. The endocrine causes of hypertension in children are generally rare. We have reviewed the diverse and rare endocrine causes of hypertension in the pediatric population. Table 3 lists features of these conditions that assist in their diagnosis. In all patients with hypertension, a thorough history and physical examination may point to the diagnosis of endocrine or other causes of secondary hypertension. For a more detailed approach to these diagnoses, other reviews may be helpful. A phased laboratory evaluation similar to that suggested by Ogborn and Crocker facilitates in the evaluation of secondary hypertension. The critical screening tests from an endocrine point of view are plasma sodium, potassium, calcium, renin activity, and thyroid function tests, including T4, T3, and thyroid stimulating hormone. Measurement of a 24-hour urine collection for aldosterone, metanephrine, and catecholamines may be warranted if the previously mentioned studies are unrevealing. More specific studies also may be suggested by these preliminary evaluations and the history and physical examination. Further investigations should be done with the additional guidance of a pediatric endocrinologist. PMID- 8417404 TI - Renal imaging in children with persistent hypertension. AB - The role that imaging plays in the evaluation of the child with hypertension depends in large part on the results of thorough historical, physical, and laboratory examinations. How aggressively one searches for an underlying renal parenchymal or renovascular disorder must be individualized in each child. An individualized approach to renal imaging in children with hypertension is presented. PMID- 8417405 TI - Childhood prevention of essential hypertension. AB - Childhood prevention of essential hypertension requires knowledge of alterable determinants of blood pressure in children; these include obesity and sodium intake and perhaps physical activity and intake of potassium and calcium. Altering these determinants may involve two general preventive strategies. The first is a population strategy, which attempts to lower blood pressure (or keep it from rising) among all children. Population strategies may require educating children to active participants in changing their behaviors (active approach) or may merely change their environment (passive approach). The second general strategy aims to focus on children at high risk of developing hypertension as adults. To determine the usefulness of this high-risk strategy, more information is needed about prediction of adult blood pressure from childhood values and about the efficacy of interventions to control blood pressure levels in high-risk children. PMID- 8417406 TI - Pharmacologic management of childhood hypertension. AB - Antihypertensive drug therapy is used in children primarily to treat secondary forms of hypertension, because the prevalence of essential hypertension in the first decade of life is considerably less than 1% of the childhood population. This prevalence increases during the second decade of life, but the percentage of teenagers with essential hypertension continues to be low. Pharmaceutical companies have been able to target drug development to specific physiologic and biochemical systems. The converting enzyme inhibitors and calcium-channel blockers have greatly improved the success of therapy concomitant with a reduction in the incidence of adverse effects. The result has been a major change during the past decade in the recommendations for antihypertensive drug therapy. PMID- 8417407 TI - Childhood predictors for high adult blood pressure. The Muscatine Study. AB - In adult populations, elevated blood pressure is related to the development of stroke, renal disease, and occlusive atherosclerosis. The significance of blood pressure levels in childhood, unless extremely elevated, has not been related to disease outcomes. In a study carried out in Muscatine, Iowa, the risk of high blood pressure in young adult life was evaluated based on the observations of blood pressure and other factors made during the school-aged years. Subjects, 2445 in number, were first observed at ages 7 through 18 years and again between 20 and 30 years. During childhood, measurements of blood pressure, height, and weight were made on alternate years. At adult ages, the same measurements were again made and a health questionnaire was administered. Adult blood pressure was correlated with childhood blood pressure, body size, and change in ponderosity from childhood to adult life. Adult ponderosity was related to childhood ponderosity, and those who were most obese as adults showed the greatest increase in weight from their childhood years. These observations suggest that strategies to prevent the acquisition of excess ponderosity during childhood may be useful in preventing adult hypertension. PMID- 8417408 TI - Relationships between blood pressure and lipids in childhood. AB - This review has discussed the potential interrelationships of elevated blood pressure and lipid abnormalities. Most of the data for this review have been collected in adults, but physiologic and genetic effects may have expression in childhood. Clustering of cardiac risk factors may be mediated by increased levels of insulin. These increased levels of insulin may be genetically determined or may be related to obesity. Recognition of the multiple risk factor phenotype is important, because individuals with multiple risk may have a much higher risk of future coronary events than do individuals with single risk factors. Medical therapy of individuals with multiple risk should be more aggressive than for individuals with single risk factors. Physicians should be cautious about therapy in the pediatric setting because of possible interaction between therapeutic agents and risk factors, however. PMID- 8417409 TI - Cardiovascular manifestations of hypertension in children. AB - Hypertension has its roots in childhood. Electrocardiographic, echocardiographic, and exercise testing in young patients point to early abnormalities of cardiovascular status in children with early hypertensive disease. Altered morphology of the cardiovascular system and vascular reactivity is also seen in the hypertensive or prehypertensive child. Perhaps investigation and preventive tactics may be best applied to the pediatric population. PMID- 8417410 TI - The renin-angiotensin system and blood pressure regulation during infancy and childhood. AB - The renin-angiotensin system plays multiple roles in the maintenance of normal blood pressure and renal function. The balance and integration of these roles change during development in ways that we do not yet fully understand. This article reviews the ways in which the renin-angiotensin system maintains normal cardiovascular homeostasis during development and its participation in physiologic and biochemical events. PMID- 8417411 TI - Adolescent obesity and hypertension. AB - Although controversy exists as to the role that insulin resistance and hyperinsulinemia play in the pathogenesis of hypertension there are ample data from both obese and nonobese subjects strongly suggesting that selective insulin resistance, hyperlipidemia, and essential hypertension are directly related. It is therefore mandatory that the evaluation of both lipid profile and insulin sensitivity needs to be made in all patients with essential hypertension. In addition it appears that childhood obesity is the single marker of the child at risk for the development of cardiovascular disease later in life. PMID- 8417412 TI - Hypertension in individuals with insulin-dependent diabetes mellitus. AB - Hypertension is known to place the individual with IDDM at high risk for the development of both renal and cardiovascular disease. Recent data suggest that aggressive antihypertensive therapy (angiotensin I converting enzyme inhibitors, prazosin, and calcium channel blockers) have significantly improved overall prognosis and long-term survival for individuals with IDDM. Because in individuals with IDDM the development of both hypertension and renal disease has its roots in childhood, it is important that early and effective antihypertensive treatment begin there. PMID- 8417413 TI - Erythromelalgia in a twenty-year-old with infectious mononucleosis. PMID- 8417414 TI - Disseminated histoplasmosis caused by Histoplasma capsulatum in an immunocompromised adolescent after exploration of a bat cave. PMID- 8417415 TI - Pseudomonas aeruginosa mastitis in a neonate. PMID- 8417416 TI - Vancomycin-associated shock in neonates. PMID- 8417417 TI - Indications for amantadine in otherwise healthy children. PMID- 8417418 TI - Respiratory syncytial virus-associated lower respiratory illnesses: possible influence of other agents. The Group Health Medical Associates. AB - Acute lower respiratory illnesses were prospectively investigated in a cohort of 1246 healthy infants who were enrolled at birth in the Tucson Children's Respiratory Study and followed through the first 3 years of life. Respiratory syncytial virus (RSV) infection was documented by culture, antigen detection or both in 276 episodes. In 21 (7.6%) of these 276, other viruses were simultaneously detected. Further serologic studies of 50 episodes in which RSV had been found increased the apparent viral codetection rate to 24%. When culture results for Chlamydia trachomatis and Mycoplasma pneumoniae were also considered, the rate of codetection was found to be 10.9% (30 of 276); this increased to 28% for the subgroup of episodes (14 of 50) that was further studied serologically. Illnesses associated with more than one agent were not significantly different from those involving RSV alone, with respect to month of onset, age at illness, illness type or duration of illness. We conclude that when RSV has been detected in previously healthy infants, routine searches for the concomitant presence of other viruses are usually not warranted. PMID- 8417419 TI - Exposure of pregnant women to ribavirin-contaminated air: risk assessment and recommendations. PMID- 8417420 TI - Factors influencing outcome in children treated with antibiotics for acute otitis media. AB - Factors affecting outcome were analyzed from 3 antibiotic clinical trials that had identical case definition and outcome criteria. Overall 102 subjects with acute otitis media had an initial tympanocentesis, were enrolled in one of the clinical trials, were randomized to receive 10 days of oral treatment and had a posttherapy visit. The antibiotics used were cefixime (38), cefaclor (25), loracarbef (14), amoxicillin plus clavulanate (16) or amoxicillin (9). Fifty-five of the 102 (54%) study subjects were classified as cured or improved at the 21- to 28-day posttherapy visit. Factors analyzed in relation to outcome included antibiotic administered, isolation of a pathogen from the middle ear aspirate, study subject age and sex, history of recurrent otitis media, unilateral vs. bilateral involvement, season of enrollment and history of antibiotic administration in the month before enrollment. Univariate analysis identified the following four factors associated with higher posttherapy visit failure rates: a history of recurrent otitis media; enrollment during winter respiratory season (December through March); a history of being treated with an antibiotic during the month before enrollment; and administration of cefaclor compared with other antibiotics. However, only a history of recurrent otitis media and enrollment during the winter respiratory season met the 0.05 significance level for entry into a model derived from logistic regression to assess interactions among factors. Clinical guidelines for the management of otitis media should take into consideration that children with a prior history or recurrent otitis media and infection during the winter season more often fail to respond to antibiotic treatment and have a higher risk of developing a persistent middle ear effusion. PMID- 8417421 TI - Comparative study of sultamicillin and amoxicillin-clavulanate: treatment of acute otitis media. AB - Sultamicillin is a mutual prodrug of ampicillin and sulbactam that is chemically linked by a diester bond. This investigational agent has beta-lactamase inhibiting activity by virtue of sulbactam, a novel beta-lactamase inhibitor. A double blind randomized study was conducted to evaluate the safety, efficacy and tolerance of sultamicillin for treatment of acute otitis media compared with amoxicillin-clavulanate. A total of 144 subjects were included (96 randomly assigned to the sultamicillin and 48 to the amoxicillin-clavulanate groups). No safety concerns for sultamicillin were identified during the study. The clinical efficacy in effusion clearance between the two groups was found not to be statistically different at 10 days (P = 0.23) and 30 days (P = 0.72). Similar rates of side effects, primarily gastrointestinal, were reported in both study groups. Sultamicillin may be an alternative for the treatment of acute otitis media when persistence and recurrence of disease become an issue. PMID- 8417422 TI - Routine tuberculin screening of children during hospitalization. AB - Harris County, TX, which includes Houston, has one of the highest childhood tuberculosis case rates in the United States. For an 11-week period in the spring of 1988 all children admitted to the medical service of the Ben Taub General Hospital in Houston, TX, received a Mantoux skin test consisting of tuberculin purified protein derivative. The purpose was to assess the impact of routine tuberculin screening during hospitalization for acute medical care and to determine whether tuberculin screening in this setting is an effective means of identifying children with asymptomatic tuberculous infection. Of the 432 patients skin tested, 50% were younger than 1 year of age and only 304 were evaluable at 48 hours. Two new positive skin tests were discovered for a positive tuberculin rate of 0.66%. We conclude that even in a high risk region, routine tuberculin screening of all children admitted to the hospital may not be effective. PMID- 8417423 TI - Sensitivity, specificity and predictive value of physical examination, culture and other laboratory studies in the diagnosis during early infancy of vertically acquired human immunodeficiency virus infection. AB - The medical records of 142 infants referred for evaluation solely because they were born to human immunodeficiency virus (HIV)-infected mothers (i.e. not because of signs or symptoms suggesting HIV infection), were reviewed. The infection status of 85 of these infants has been determined; 17 (20%) have confirmed HIV infection and 68 have seroreverted to HIV and lack evidence of infection. During the first 6 months of life HIV culture had better sensitivity, specificity, positive predictive value and negative predictive value for diagnosis of HIV infection than did physical examination, serum immunoglobulin determination or HIV p24 antigen determination. Of the 16 HIV-infected infants who were available for evaluation during the first 6 months of life, all had at least one culture from blood positive for HIV. Two of 4 and 10 of 11 infants were culture-positive at birth and during the first 3 months of life, respectively. A positive HIV culture results was the earliest finding of infection in 15 infants; 10 of these infants concomitantly were found to have hyperimmunoglobulinemia (8 cases) and/or an abnormal physical examination (4 cases). One HIV-infected infant developed hyperimmunoglobulinemia G and A at age 3 months without other evidence of HIV infection until age 5 months when a positive HIV culture was noted. All HIV-infected infants had abnormal findings by physical examination, a positive HIV culture, and/or hyperimmunoglobulinemia by 3 months of age. Infants with normal physical examination and laboratory test results at 3 and/or 6 months of age invariably were HIV-uninfected. PMID- 8417424 TI - Initial clinical assessment of the comatose patient: cerebral malaria vs. meningitis. AB - One hundred twenty-one Liberian children were admitted in coma to the ELWA Hospital, Monrovia, Liberia. Admitting diagnoses, before lumbar puncture, were compared with discharge diagnoses. Ninety-four children were discharged with a final diagnosis of cerebral malaria and 27 with a diagnosis of meningitis. The admitting diagnosis was correct in 76.6% (72 of 94) of patients with cerebral malaria and 59.3% (16 of 27) of patients with meningitis. The cerebrospinal fluid leukocyte count was the single most significant factor in determining the correct diagnosis. Without the cerebrospinal fluid analysis, the discriminant accuracy (77%), i.e. definitive separation of the two illnesses, was comparable to the physician's admission diagnosis (73%). Other data contributing to the differential diagnosis of cerebral malaria and meningitis included the number of days of fever before admission, the presence or absence of nuchal rigidity, fontanelle fullness and peripheral blood malaria smear. Mortality rates for cerebral malaria and meningitis were 14.9 and 29.6%, respectively. These data suggest that physicians cannot reliably discriminate between cerebral malaria and meningitis without cerebrospinal fluid analysis. PMID- 8417425 TI - Epidemic measles in the postvaccine era: evaluation of epidemiology, clinical presentation and complications during an urban outbreak. AB - A total of 440 cases of measles were seen at a single children's hospital during an epidemic in a large urban center. These cases were retrospectively analyzed for demographic characteristics, exposure histories, risk factors for and incidence of complications, outcome and financial consequences. More than 90% of children were < 5 years of age and 56% were < 15 months of age. Ninety-four percent were of Hispanic or African-American ethnic background. A visit to a medical facility was the only source of infection for 83 of the 115 patients for whom exposure histories were available. Four patients with underlying immunodeficiency were among those nosocomially exposed and one died of pulmonary complications. Of the 440 patients 195 (44.3%) required hospitalization for measles complications. The mean age of hospitalized patients, 1.9 +/- 2.4 years, was not different from those not admitted (2.1 +/- 2.5 years; P < 0.25). The most frequent indications for admission were respiratory tract complications (pneumonia and/or croup) seen in 161 patients and dehydration in 25 patients. Fourteen required intensive care and 11 required intubation and mechanical ventilation. Three children died, all of pulmonary involvement. Total hospital and medical charges for care of these patients was more than $1.7 million dollars. PMID- 8417426 TI - Outbreak of leptospirosis associated with swimming. AB - Between July 7 and 18, 1991, five boys from a small town in rural Illinois experienced the onset of an acute febrile illness subsequently confirmed as leptospirosis by serologic tests. A cohort study found that swimming in a small swimming hole, Steel Tunnel Pond, was associated with disease (P < 0.01), the attack rate being 28%. Leptospira interrogans serovar grippotyphosa was isolated from urine cultures from two of the case patients and from a culture of Steel Tunnel Pond water. A high seroprevalence for grippotyphosa was found in animals near the pond. Drought conditions had been present in the month before the outbreak, creating an environment in the pond which probably facilitated transmission of the organism from area animals to humans. Although leptospirosis is infrequently reported in humans in the United States, it is endemic in animals and the potential for outbreaks exists, especially when environmental conditions are favorable. PMID- 8417427 TI - Management of acute diarrhea in children: lessons learned. PMID- 8417428 TI - Microbiologic and clinical aspects of a trial of once daily cefixime compared with twice daily cefaclor for treatment of acute otitis media in infants and children. AB - In this randomized, investigator-blinded multicenter study, tympanocentesis for acute otitis media with effusion in 137 ears from 108 children, 6 months to 12 years of age, revealed 84 definite pathogens and 32 potential pathogens. Twenty nine aspirates from 23 subjects were sterile. Of the 116 isolates 42 (36%) were Streptococcus pneumoniae, 24 (21%) were Haemophilus influenzae, 9 (8%) were Moraxella catarrhalis, 9 (8%) were Streptococcus pyogenes and 1 (1%) was Staphylococcus aureus. Twenty-two (19%) definite pathogens produced beta lactamase. Patients were randomized to cefixime (8 mg/kg/day daily) or cefaclor (40 mg/kg/day divided into two doses). Efficacy was determined by pneumatic otoscopy and tympanometry at the end of therapy visit on Days 11 to 14 and up to 4 weeks of follow-up. At end of therapy subjects with definite pathogens exhibited a satisfactory clinical outcome in 26 of 36 (72%) ears for cefaclor and 40 of 48 (83%) ears for cefixime recipients (P = 0.12). For ears with beta lactamase-producing isolates there were no (0 to 12) cefixime failures but 4 of 10 cefaclor failures (P = 0.03). Diarrhea/loose stools were more frequent in cefixime (16 of 58) than cefaclor (4 of 50) recipients. One cefixime subject required discontinuation of drug. Overall efficacy for treatment of acute otitis media with effusion was not different; however, cefixime appeared more effective for infections caused by beta-lactamase-producing organisms. PMID- 8417429 TI - Cefixime vs. cefaclor in the treatment of acute otitis media in children: a randomized, comparative study. AB - The efficacy of cefixime was compared with that of cefaclor in the treatment of 63 patients with acute otitis media. Patients received either a single dose of cefixime (8 mg/kg/day) or 3 divided doses of cefaclor (40 mg/kg/day). On the basis of otoscopic and tympanometric results at 10 to 14 days after the start of treatment, 28 (97%) of 29 cefixime-treated patients and 25 (78%) of 32 cefaclor treated patients had resolution of acute otitis media. The clinical cure rate associated with all organisms was 94% for cefixime (16 of 17 isolates) and 68% (13 of 19 isolates) for cefaclor. The cure rate for Streptococcus pneumoniae was 12 of 12 (100%) for cefixime and 7 of 7 (100%) for cefaclor; the cure rate for Haemophilus influenzae (which includes 2 patients with mixed infections) was 3 of 4 (75%) for cefixime and 2 of 7 (29%) for cefaclor. One clinical relapse occurred among 29 cefixime-treated patients; however, at 28 days 9 recurrences were observed. Three of 25 (9%) cefaclor-treated patients failed and 4 (13%) relapsed at 10 to 14 days, an additional 2 (10%) experienced recurrence by Day 28. Eight (28%) cefixime-treated patients experienced adverse events (7 gastrointestinal and 1 diarrhea and rash); 8 (25%) cefaclor-treated patients experienced adverse events (all gastrointestinal). Our data suggest that both at end of therapy and for 14 days thereafter, cefixime given once a day for acute otitis media is clinically equivalent to cefaclor given 3 times a day. PMID- 8417430 TI - Review of cefixime in the treatment of otitis media in infants and children. AB - During the past decade there has been an increase in the percentage of resistant bacteria isolated from middle-ear effusions aspirated from infants and children who have had acute otitis media. At least nine oral antibiotics or combination agents are available for this indication. Cefixime, a third generation cephalosporin, has excellent in vitro activity against both beta-lactamase negative and beta-lactamase-positive Haemophilus influenzae and Moraxella catarrhalis, good activity against Streptococcus pneumoniae and Streptococcus pyogenes but relatively poor activity against Staphylococcus aureus. In children cefixime is similar in effectiveness to amoxicillin and cefaclor, but diarrhea and stool changes are more common with cefixime. Amoxicillin is still preferred for initial empiric treatment of uncomplicated acute otitis media. Its major drawback is limited efficacy when beta-lactamase-producing bacteria are the causative organisms. Cefixime is a viable alternative to amoxicillin for infants and children with acute otitis media when: (1) a beta-lactamase-producing H. influenzae or M. catarrhalis is isolated from otorrhea or tympanocentesis; (2) the child has a history of delayed hypersensitivity to the penicillins but no history of hypersensitivity to the cephalosporins; (3) there is a high incidence of resistant bacteria in the community; (4) there is not clinical improvement with amoxicillin; or (5) once daily administration is more convenient. PMID- 8417431 TI - The acute phase host reaction during bacterial infection and its clinical impact in children. PMID- 8417432 TI - Management of acute hematogenous osteomyelitis and septic arthritis in the pediatric patient. PMID- 8417433 TI - Enhanced p24 antigen detection in sera from human immunodeficiency virus-infected children. PMID- 8417434 TI - Early diagnosis of human immunodeficiency virus type 1-infected infants by plasma p24 antigen assay after immune complex dissociation. PMID- 8417435 TI - Chylous ascites caused by Mycobacterium avium complex and mesenteric lymphadenitis in a child with the acquired immunodeficiency syndrome. PMID- 8417436 TI - Day care in America: what is needed? PMID- 8417437 TI - Looking at the child care landscape. PMID- 8417438 TI - Quality of child care as an aspect of family and child care policy in the United States. PMID- 8417439 TI - Child care for children with special needs. PMID- 8417440 TI - Day care for the child with acquired immunodeficiency syndrome and the child of the drug-abusing mother. PMID- 8417441 TI - Continuity of supervised care for school-age children. PMID- 8417442 TI - Employers and child care. PMID- 8417443 TI - The pediatrician's role in helping parents with out-of-home child care. PMID- 8417444 TI - Impact of day care on parents and family. PMID- 8417445 TI - Impact of day care on the child. AB - In the world of day-care research, the status of our knowledge is sufficiently shaky that we must continue to keep an open mind about the service. The knowledge base is growing rapidly, but the conceptual structure that supports it is flimsy and insubstantial. Fortunately, current research efforts are improving this situation. Regardless of whether we like or dislike day care, it is, like the family, here to stay. That realization alone should strengthen our resolve not to compromise on the type of service we create. We have to continue to identify parameters of quality and become good matchmakers in terms of child care, family, and child characteristics. Through such efforts, a network of educare programs that will foster favorable development in children can become a national and global reality. PMID- 8417446 TI - Care of the ill child in day-care settings. PMID- 8417447 TI - Day-care standards: need and impact. PMID- 8417448 TI - The pediatrician's role in setting standards of day care for children. PMID- 8417449 TI - Role of the pediatrician in setting and using standards for child care. PMID- 8417450 TI - Ensuring quality, affordable child care: mobilizing for action. PMID- 8417451 TI - International perspectives on child care policies and programs. PMID- 8417452 TI - Alternative child care in France: advances in the study of motor, interactive, and social behaviors of young children in settings allowing them to move freely in a group of peers. AB - This paper reports new data on the emergence and development from the 9th to 19th month of age of motor, interactive, and social behaviors of 17 children with peers in a milieu that allows the "conquest" of the third dimension of space without limitation of their motor activity. Their motor skills and social behaviors appear more precocious and complex than previously indicated by scientific publications, clinical observations, and developmental scales. 1. The motor skills that allow the "conquest" of climbing structures (ascents of a spiral staircase and a ramp) are already observed before the age of 12 months in some children (ascents of the ramp) or nearly half of them (ascents of the staircase). Complex social behaviors (multimodal interactions from here and there along a holed partition, imitations, cooperations, anticipation of the behavior of the partner) also emerge before 12 months. 2. The age range from 12 to 14 to 15 months is characterized by consolidation of the motor skills that allow the "conquest" of the third dimension of the space and a rapid increase of the imitations and interactions around the holed partition. 3. The age range from 14 to 18 months is characterized by diversification of the posturomotor patterns during ascents and descents of staircase and ramp and the growing predominance of the standing position. This is also a phase of significant increase in frequency of imitations and cooperations and of emergence of complex forms of cooperation. 4. Aggression is infrequent at all ages. PMID- 8417453 TI - Scandinavian experience in providing alternative care. PMID- 8417454 TI - Putting a child in day care: issues for working parents. PMID- 8417455 TI - Endurance training of trunk extensor muscles. AB - BACKGROUND AND PURPOSE: The purpose of this study was to assess the effects of an endurance exercise protocol on the isometric holding time of the trunk extensor muscles (mechanical fatigue measure) and on the recorded median frequency (MF) measurements from the surface electromyogram (physiological fatigue measure). SUBJECTS: Twenty-eight healthy female volunteers were selected to participate in the study. METHODS: The subjects were stratified by activity level and then assigned to an exercise or a control group and tested at weeks 0, 3, and 6. Reliability (r) of the measurements, established by testing each subject twice at week 0, was .87. Exercise group subjects trained at home twice daily for 6 weeks, progressing by established protocol. RESULTS: The exercise group subjects increased their isometric holding time by 22% after 6 weeks. The control group showed no significant changes. CONCLUSION AND DISCUSSION: The home program was believed to be effective for increasing isometric endurance of the trunk extensors, measured by holding time. The MF measurements prior to and just after a fatiguing contraction remained stable over time. The change in the slope of the MF with fatigue did not reach statistical significance. This physiologic measure of fatigue did not show commensurate changes with training in this group of subjects. Possible reasons for the results are discussed. PMID- 8417456 TI - The impact of the prospective payment system: perceived changes in the nature of practice and clinical education. AB - BACKGROUND AND PURPOSE: Health care financing for teaching hospitals has undergone significant change in the past decade. This report describes changes in physical therapy practice and clinical education in three New England hospitals from 1984 to 1988. SUBJECTS: Hospital administrators, physical therapy managers, and clinical educators (N = 18) from the three teaching hospitals participated in this descriptive study. METHODS: Demographic, environmental, and participant interview data were gathered and examined to identify changes during this period. RESULTS: Perceived changes in practice include growth in specialized knowledge; increased emphasis on health care quality, efficiency, and accountability; new ethical dilemmas for practitioners; and a changing physical therapy role with new professional development opportunities. Perceived changes associated with clinical education were increased student performance expectations, unchanged resources for clinical education, greater emphasis on student self-directedness, and continued high valuing of this setting for physical therapy clinical education. CONCLUSION AND DISCUSSION: These results indicate significant change in the role of the physical therapist within these settings and suggest how these changes influence the clinical education of physical therapy students in these teaching hospitals. PMID- 8417457 TI - Reliability of the modified-modified Schober and double inclinometer methods for measuring lumbar flexion and extension. AB - BACKGROUND AND PURPOSE: The primary purpose of this study was to determine the reliability of lumbar flexion and extension range-of-motion measurements obtained with the modified-modified Schober and the double inclinometer methods on subjects with low back pain. SUBJECTS: Fifteen patients (8 women, 7 men), aged 25 to 53 years (mean = 35.7, SD = 9.9), with chronic low back pain were measured by three physical therapists with 3 to 12 years (mean = 8.3, SD = 4.7) of clinical experience. METHODS: The therapists used the modified-modified Schober and double inclinometer techniques to measure, in random order and on two occasions, the subjects' lumbar flexion and extension. RESULTS: Pearson Product-Moment Correlation Coefficients for test-retest reliability for the modified-modified Schober technique varied from .78 to .89 for lumbar flexion and from .69 to .91 for extension; for the double inclinometer method, Pearson correlation coefficients varied from .13 to .87 for lumbar flexion and from .28 to .66 for extension. Analysis of variance-derived intraclass correlation coefficients for interrater reliability for the modified-modified Schober technique were .72 for flexion and .76 for extension; for the double inclinometer technique, they were .60 for flexion and .48 for extension. CONCLUSION AND DISCUSSION: The modified modified Schober method thus appears to be a reliable method for measuring lumbar flexion and extension for patients with low back pain, whereas the double inclinometer technique needs improvement. PMID- 8417458 TI - The effect of training on physical therapists' ability to apply specified forces of palpation. AB - BACKGROUND AND PURPOSE: The aim of this study was to evaluate whether postgraduate physical therapy students studying manipulation could learn to accurately produce specific forces during palpation of an intervertebral joint. SUBJECTS: The 12 subjects (7 female, 5 male), aged 26 to 36 years (X = 29.5, SD = 2.9), had each completed a 4-year degree course in physical therapy and had worked between 3 and 10 years in clinical practice. All subjects were enrolled in a 12-month postgraduate manipulative therapy diploma course. METHODS: Subjects in the experimental group (n = 6) trained to apply specific forces of 1, 5, 10, 15, 20, and 25 kiloponds using bathroom scales. They practiced for 10 minutes per day for 30 days. Their ability to produce these forces on command was measured using a force platform as they applied posteroanterior passive accessory intervertebral joint movements to the lumbar spine of the healthy subjects. This testing was done prior to training (pretest), immediately after training (posttest), and 1 month following cessation of training (retention test). The control group subjects (n = 6) had no training with scales but were also students of the postgraduate manipulative physical therapy course. RESULTS: In comparison with the control group, the experimentally trained group showed reduced error in force production both immediately after training and 1 month later. This improvement was significant for the retention test. For the retention test, the experimental group subjects were also tested on the trained task (ie, their ability to apply specific forces to the scales). They developed higher levels of accuracy than did the control group. CONCLUSION AND DISCUSSION: Experimental training, therefore, was an effective addition to normal training, suggesting that therapists can learn to quantify applied forces, with significant implications for communication and evaluation of joint behavior. PMID- 8417459 TI - Fact or ...? PMID- 8417460 TI - TENS--an international perspective. PMID- 8417461 TI - Deyo clarifies position. PMID- 8417462 TI - A disease of the spirit. PMID- 8417463 TI - Strokes involving gray matter: studies on in situ models of cerebral ischemia. PMID- 8417464 TI - Anoxic injury of central myelinated axons: ionic mechanisms and pharmacology. PMID- 8417465 TI - The ischemic penumbra in stroke: prospects for analysis by nuclear magnetic resonance spectroscopy. PMID- 8417466 TI - The therapeutic window for methylprednisolone treatment of acute spinal cord injury: implications for cell injury mechanisms. PMID- 8417467 TI - Emerging strategies for the treatment of ischemic brain injury. PMID- 8417468 TI - The design and appraisal of randomized clinical trials in cerebrovascular disease and CNS trauma. PMID- 8417469 TI - Current antisense nucleic acid strategies for manipulating neuronal and glial cells. PMID- 8417470 TI - Factors affecting proviral expression in primary cells grafted into the CNS. PMID- 8417471 TI - Cellular replacement of dopamine deficit in Parkinson's disease using human fetal mesencephalic tissue: preliminary results in four patients. PMID- 8417472 TI - Intracerebral transplantation: prospects for neuronal replacement in neurodegenerative diseases. PMID- 8417473 TI - Mitochondrial dysfunction as a mechanism of CNS injury. PMID- 8417474 TI - Laparoscopic general surgery--past, present, and future. PMID- 8417475 TI - Hepatectomy with initial suprahilar control of intrahepatic portal pedicles. AB - In 1985 Couinaud described an original technique for left hepatectomy. This technique is based on initial control of the left intrahepatic portal pedicle after wedge incision of the hepatic capsule is made and the hilar plate is lowered before the hepatic parenchyma is opened. The vascular and biliary pedicular elements are dissected concomitantly at a point where they are protected by and contained within a sheath emanating from the hepatic capsule. After elective clamping of the pedicle at this level is done, the territory to be removed becomes obvious by the change of color produced by ischemia, and bleeding is minimized preventively. Although Couinaud's technique concerned only one case of left hepatectomy, we confirmed the ease, safety, and reproducibility afforded by this technique in four left hepatectomies. Based on the same principles, this technique may be used for other types of hepatic resections, notably, left hepatectomy extended to segments 5 and 8 (trisegmentectomies). We report our experience with this technique in 15 cases of major hepatic resections. PMID- 8417476 TI - Graft replacement of pararenal inferior vena cava for leiomyosarcoma with the use of venous bypass. AB - Total replacement of the inferior vena cava across the renal vein confluence was successfully performed under venovenous bypass in a patient with primary leiomyosarcoma of the inferior vena cava. Concomitant resection of metastases by right nephrectomy, partial hepatectomy, and cholecystectomy allowed recurrence free survival for the past 20 months. To the best of our knowledge, this is the first report of successful replacement of the inferior vena cava for a primary tumor. PMID- 8417477 TI - Neutropenic enterocolitis in patients without leukemia. AB - BACKGROUND: Neutropenic enterocolitis is a serious complication involving the bowel in patients with severe neutropenia. Most of the patients are undergoing chemotherapy for acute leukemia. METHODS: This is a report of three cases of nonleukemic neutropenic enterocolitis. In these cases neutropenia was caused by an analgesic (dipyrone), chemotherapy for multiple myeloma, and chemotherapy for lung carcinoma. Two patients were treated by surgery, one medically. RESULTS: The patient with multiple myeloma who was treated by exteriorization of the sigmoid colon died on the tenth postoperative day of multiple organ failure caused by sepsis. The other two patients survived. CONCLUSIONS: A review of the literature revealed 14 other patients without leukemia who had neutropenic enterocolitis. The cause, diagnosis, and possible treatment options of this entity are discussed. PMID- 8417478 TI - American Board of Surgery In-Training/Surgical Basic Science Examination (ABSITE) comes of age. PMID- 8417479 TI - Venous disease, injury, and politics. PMID- 8417480 TI - Colonic ischemia after an abdominal aortic aneurysm (AAA) repair through a retroperitoneal approach. PMID- 8417481 TI - Isolated insufficiency of primary perforator veins. PMID- 8417482 TI - Re: Surgical management of BANS malignant melanoma. PMID- 8417483 TI - Accuracy of endoscopic ultrasonography in the diagnosis and staging of gastric cancer and lymphoma. AB - BACKGROUND: There is a need to assess the sensitivity, specificity, and predictive value of endoscopic ultrasonography (EUS) in the diagnosis and staging of gastric cancer and lymphoma. METHODS: A prospective study was performed on 86 patients with endoscopic gross appearance suspicious for cancer or lymphoma. Biopsies with endoscopic forceps were always carried out before EUS. All patients underwent laparotomy for final diagnosis, staging, and eventually treatment. The results of EUS were correlated with the histologic findings of the resected specimens, when possible, or with the surgical findings. There were 42 gastric cancers and 44 primary gastric lymphomas. RESULTS: EUS made a correct diagnosis of cancer in 35 of 42 patients, with a sensitivity of 83%. Positive predictability was 87%, specificity was 97%, and negative predictability was 96%. Diagnostic accuracy was 95%. In the evaluation of cancer depth invasion, EUS was correct in 91% of cases. EUS displayed perigastric metastatic lymph nodes in 14 of 25 patients, with a sensitivity of 56%. Positive predictive value was 93%, specificity was 93%, and negative predictive value was 54%. Diagnostic accuracy was 69%. EUS made a correct diagnosis of lymphoma in 39 of 44 patients, with a sensitivity of 89%. Positive predictability was 87%, specificity was 97%, and negative predictability was 97%. Diagnostic accuracy was 95%. In the evaluation of lymphoma depth invasion, EUS was correct in 92% of cases. EUS displayed metastatic perigastric lymph nodes in 8 of 18 patients, with a sensitivity of 44%. Positive predictability was 100%, specificity was 100%, and negative predictability was 72%. Diagnostic accuracy was 77%. CONCLUSIONS: From these data it appears that in these diseases EUS has demonstrated specific ultrasonographic features that allow correct diagnosis and staging in the majority of patients. In difficult cases EUS may help to achieve the correct diagnosis. EUS also appear to be a useful tool for staging of gastric cancer and lymphoma. It shows not only tumor depth and local spread but also the passage from a pathologic to a normal wall and lymph node metastasis. With this accurate noninvasive staging procedure, in the near future many patients will no longer undergo exploratory laparotomy for surgical staging. Thanks to EUS, the choice of conservative or surgical treatment can be strongly affected. In case of surgery, EUS can orient the kind of surgical approach. Moreover, the use of EUS for evaluation of therapy during follow-up will probably become of major importance. PMID- 8417484 TI - Lovastatin and gallstone dissolution: a preliminary study. AB - BACKGROUND: Lovastatin, an agent that reduces both serum and biliary cholesterol in humans, also inhibits cholesterol gallstone formation in an animal model. The present study was designed to assess the efficacy of lovastatin in gallstone dissolution. METHODS: All prairie dogs were fed a 1.2% cholesterol-enriched diet during the entire study. Gallbladders from five animals were examined at 3 weeks, and four of five gallbladders contained gallstones. Remaining animals were maintained on the 1.2% cholesterol-enriched diet and randomized to receive either water (n = 7); lovastatin, 8 mg (n = 7); ursodeoxycholic acid, 50 mg (UR, n = 7); or both drugs (lovastatin and UR, n = 7) twice daily by way of orogastric tube for 4 additional weeks. Response to therapy was determined by blinded examination of gallbladders. RESULTS: All three treatment groups had significant reductions in serum cholesterol, hepatic bile cholesterol, and hepatic cholesterol saturation index as compared to controls (water). Lovastatin induced a 28% response rate to dissolution therapy, which was equal to that achieved with UR, and the combination of lovastatin and UR produced a 56% response rate. CONCLUSIONS: This preliminary study suggests that lovastatin, alone or in combination with UR, may be useful in dissolving gallstones in humans. PMID- 8417485 TI - Acceleration of tissue repair by transforming growth factor beta 1: identification of in vivo mechanism of action with radiotherapy-induced specific healing deficits. AB - BACKGROUND: Transforming growth factor beta 1 (TGF-beta 1) is an effective accelerator of soft tissue repair in both normal and impaired healing models; however, its in vivo mechanism of action remains unclear. Modern radiation techniques can create unique healing deficits, allowing for a more specific definition of tissue response to growth factor therapy. In the rat linear skin incision model, cobalt 60 photon beam total body irradiation (TBI), 800 rads, causes a marked depression of circulating monocytes and largely spares the skin tissue. Megavoltage electron beam surface irradiation (SI), 2500 rads, markedly impairs surface healing while sparing the bone marrow. With these models of selective healing deficits, the ability of TGF-beta 1 to accelerate tissue repair directly in the absence of circulating macrophage precursors (TBI) or in the presence of damaged dermal fibroblasts (SI) was evaluated. METHODS: Adult male Sprague-Dawley rats were randomly assigned to groups of TBI, SI, or nonirradiated sham controls and received radiation 2 days before wounding. Paired linear full thickness skin incisions were created and a single dose of TGF-beta 1 (2 micrograms/wound) or vehicle control was applied to each wound. RESULTS: Both radiation techniques produced a marked healing deficit when assessed on postwounding days 7 and 12. TBI treatment was characterized by severe monocytopenia, confirmed by a tissue macrophage-specific immunohistochemical technique. On days 7 and 12 after wounding, TGF-beta 1 significantly accelerated soft tissue repair and wound-breaking strength in the TBI-treated rats, demonstrating an ability to directly promote the induction of collagen synthesis in the absence of monocytes/macrophages. In contrast, TGF-beta 1 was unable to reverse the SI-induced healing deficit characterized by impaired function of dermal fibroblasts. CONCLUSIONS: These in vivo observations provide further evidence for a direct mechanism of action by TGF-beta 1 on collagen synthesis by wound fibroblasts during soft tissue repair. PMID- 8417486 TI - Surgical management of short bowel syndrome. PMID- 8417487 TI - Duodenogastric reflux quantification in peptic ulcer surgery: comparison between different surgical techniques. AB - BACKGROUND: We quantified duodenogastric reflux with 6-hour continuous intravenous infusion of technetium 99m-labeled hepatoiminodiacetic acid (99mTc HIDA) and subsequent quantification in gastric juice. METHODS: For this purpose, 50 patients were studied who had undergone surgery on the stomach with different surgical techniques: bilateral vagotomy plus Heineke-Mikulicz pyloroplasty, bilateral truncal vagotomy plus anterior pylorectomy, proximal gastric vagotomy, antrectomy and Billroth I reconstruction, and antrectomy and Billroth II reconstruction, comparing them with 10 healthy subjects used as a control group. We also studied the existing correlation between the rates of reflux determined by 99mTc-HIDA and those of total bile acids in gastric juice. RESULTS AND CONCLUSIONS: Patients who underwent gastric surgery had significantly greater quantities of duodenogastric reflux (p < 0.001) than had the control group. When the groups undergoing gastric surgery were compared, the patients who underwent resection showed higher reflux rates (p < 0.001) than did the patients who did not undergo resection. We found no differences among the groups of patients who did or did not undergo resection. We also found a highly significant correlation (p < 0.001) between the concentrations of 99mTc-HIDA and bile acids in gastric juice. PMID- 8417488 TI - Isolation and long-term culture of human hepatocytes. AB - Pressure-controlled perfusion of specimens of surgical hepatic resection produced improved yields of human hepatocytes for studies of long-term cultures. The effect of an extracellular matrix configuration on albumin secretion was evaluated by culture on a single layer of collagen or between double layers of collagen gel with Dulbecco's modified Eagle and Williams E media. Hepatocytes from 12 patients were maintained for more than 30 days, and in five of 12 experiments cells were cultured beyond 2 months. In the double gels the cells demonstrated typical polygonal liver cell morphology and higher albumin secretion (p < 0.01) up to 65 days; in contrast, in the single gels cells spread horizontally, and albumin secretion declined rapidly within 3 weeks. A comparison of media formulations showed that maximum albumin secretion occurred 5 days later but was maintained significantly longer with Dulbecco's medium (p < 0.01). The simple addition of a second layer of gelled collagen forming a collagen sandwich significantly stabilizes and supports the long-term culture of human hepatocytes. PMID- 8417489 TI - Four-compartment fasciotomy and venous calf-pump function: long-term results. AB - Twenty-one patients who underwent a four-compartment fasciotomy between January 1, 1986, and December 31, 1987, were revisited with respect to venous calf-pump function 32 to 56 months (mean, 46.2 months) after fasciotomy. Enrollment criteria consisted of an intact motor innervation, palpable pedal pulses, the absence of venous hypertension, and deep venous thrombosis before and during the treatment requiring fasciotomy and unimpaired ankle and knee joint function. None of the patients showed signs of chronic venous hypertension at assessment. Ambulatory strain-gauge plethysmography revealed no significant difference in recovery time and refilling volume values between the two limbs of each patient (p > 0.1) and between limbs in which fasciotomy was performed and those of normal subjects (p > 0.1). Color flow duplex scanning revealed patent popliteal veins with normal respiration-induced phasic flow pattern and absent reflux in all patients. All visible calf veins were patent, compressible, and without morphologic alterations. The fasciotomy wound was closed by delayed skin suture (the fascia remaining divided) in 48% and by skin grafts of the lateral incision in 52% of the patients, without significant difference in recovery time and refilling volume values (p > 0.1) at assessment. Fasciotomy does not lead to venous calf-pump dysfunction, irrespective of whether the wound is closed by delayed suture or skin grafts. PMID- 8417490 TI - Long-term results of entry closure and aneurysmal wall plication with axillofemoral bypass: a new procedure for repair of DeBakey type 3 dissecting aneurysm. AB - To prevent aneurysm rupture, avoid pseudoaneurysm formation, and preserve intercostal arteries, a new procedure for repair of DeBakey type 3 dissecting aneurysm was developed. Since January 1977, 28 patients have undergone repair of type 3 dissecting aneurysm. Fifteen patients with type 3b underwent this new procedure (group 1) and 13 patients with type 3a dissecting aneurysm underwent segmental graft replacement (group 2). In group 1 a permanent axillofemoral bypass was placed on the right side. Next the intrathoracic false lumen was opened longitudinally, the entry was closed, and the aneurysmal wall was sutured around the true lumen as tightly as possible. The operative mortality rate was 20% in group 1 and 31% in group 2. One of 15 patients in group 1 died of operation-related causes, whereas three patients in group 2 died. There were six late deaths: three in group 1 and three in group 2. Paraplegia occurred in neither group 1 nor group 2. The mean diameter of the plicated descending aorta was 24.0 +/- 2.7 mm 3 months after surgery. No recurrence was detected in group 1. These results suggested that this new surgical technique for repair of type 3 dissection reduces the incidence of paraplegia and pseudoaneurysm formation. PMID- 8417491 TI - Technique and clinical results of endorectal surgery. AB - At the Surgical Department of the University of Cologne, a new system has been developed for transanal endoscopic surgery that allows all the conventional surgical techniques within the entire rectal cavity. The method has been in clinical use since 1983. The main indication is the removal of sessile adenomas. Early rectal carcinomas with favorable histologic grading (grades 1 and 2) and staging (Mason I and II) are also suitable for the method. Advanced cancers can also be removed endoscopically in one session, but we perform local excisions of advanced cancers only in cases in which the patient is unwilling to undergo extensive surgery. During the period from July 1983 to December 1990, this method has been employed on 233 patients in 251 cases. The intraoperative and postoperative course of 236 (94%) operations out of 251 was free from any complications. Early postoperative complications consisted of intraperitoneal perforations (five cases), rectovaginal fistulas (four cases), hemorrhages (four cases), and death as a result of cardiopulmonary failure (two cases). The recurrence rate of adenomas is 4.9%. Endorectal surgery allows endoscopic local transanal excision of large adenomas and early cancers with minimal morbidity and excellent presentation of specimens for complete histologic analysis. PMID- 8417492 TI - Protective effect of platelet-activating factor antagonist on ischemia-induced liver injury in rats. AB - Platelet-activating factor (PAF), one of the chemical mediators related to inflammation reaction, is also involved in the pathologic state induced by endotoxin or ischemia. PAF antagonist has been reported to block the action of PAF and protect cells from its deleterious effects. The effects of a PAF antagonist, CV-6209, were evaluated in this study by means of a partial liver ischemia model, in which ischemia was induced by clamping only part of the liver without causing intestinal congestion. This model allowed the study of ischemic liver injury without influence from other organs. After 30, 60, and 90 minutes of ischemia, the bile flow, ATP level, and energy charge of the ischemic lobes were compared for the effects with and without CV-6209. After 60 minutes of ischemia, those that had received CV-6209 showed more bile production and higher ATP level and energy charge, with values of 0.25 +/- 0.05 ml/hr, 3.9 +/- 0.9 nmol/mg dry liver weight, and 0.61 +/- 0.02, respectively. In contrast, the values for the control group were 0.05 +/- 0.05 ml/hr, 1.7 +/- 0.8 nmol/mg dry liver weight, and 0.43 +/- 0.08, respectively. Other liver function tests (aspartate aminotransferase and lactate dehydrogenase levels) could also be improved if an appropriate dose of PAF antagonist were administered. The results imply that PAF, as has been suggested in other studies on ischemic injury, plays a role in liver ischemia and that its deleterious effects can be blocked by PAF antagonist. We conclude that the PAF antagonist offers promise in the field of liver surgery, including liver transplantation, as a means of protecting the liver from ischemic injury. PMID- 8417493 TI - What does the American Board of Surgery In-Training/Surgical Basic Science Examination tell us about graduate surgical education? AB - BACKGROUND: This research sought to identify the strengths and weakness in residents' basic science knowledge and, second, to determine whether they progressively improve in their abilities to recall basic science information and clinical management facts, to analyze cause-effect relationships, and to solve clinical problems. METHODS AND RESULTS: Basic science knowledge was assessed by means of the results of the January 1990 American Board of Surgery's In Training/Surgical Basic Science Exam (IT/SBSE). Postgraduate year (PGY) 1 residents' scores were compared with those of PGY5 residents. Content related to a question was considered "known" if 67% or more of the residents in each of the two groups answered it correctly. Findings showed 44% of the content tested by the basic science questions were unknown by new and graduating residents. The second research question required the 250 IT/SBSE questions to be classified into one of three levels of thinking abilities: recall, analysis, and inferential thinking. Profile analysis (split-plot analysis of variance) for each pair of resident levels indicated significant (P < 0.001) differences in performance on questions requiring factual recall, analysis, and inference between all levels except for PGY3s and PGY4s. CONCLUSIONS: The results of this research enable program directors to evaluate strengths and weaknesses in residency training curricula and the cognitive development of residents. PMID- 8417494 TI - The effect of octreotide on hepatic regeneration in rats. AB - The effect of the long-acting somatostatin analog octreotide on liver regeneration was studied in rats in vitro and in vivo. The effect of continuous subcutaneous octreotide infusion on regenerative liver weight and relative DNA synthesis was examined in rats that had undergone 70% hepatectomy. Administration of octreotide resulted in a 33% reduction of regenerating liver weight at 72 hours and a 67% reduction of regenerative hepatocellular hyperplasia at 24 hours. This effect was reversed within 12 hours after withdrawal of the drug. The mechanism for the inhibitory effect of octreotide appears to be indirect, because experiments in hepatocyte cultures did not demonstrate a direct inhibitory effect on serum-free or epidermal growth factor-induced regenerative hepatocyte proliferation. Because insulin levels were suppressed by octreotide in the in vivo experiments, suppression of hepatotrophs may be the mechanism by which octreotide inhibits liver regeneration. PMID- 8417495 TI - The mechanism of conversion of xanthine dehydrogenase to xanthine oxidase in acute pancreatitis in the canine isolated pancreas preparation. AB - Four models of acute pancreatitis have been previously developed that use the ex vivo perfused isolated canine pancreas preparation. The four models include the intraarterial infusion of oleic acid (FFA) that mimics hyperlipemic pancreatitis, partial obstruction of the pancreatic duct with secretin stimulation (POSS) that mimics gallstone pancreatitis, a 2-hour period of ischemia before perfusion (ISCH 2) that mimics shock pancreatitis, and the infusion of cerulein at supramaximal stimulatory doses (CER), which lacks an obvious clinical counterpart. In the FFA, POSS, and ISCH 2 pancreatitis, but not in the CER pancreatitis, toxic oxygen metabolites, generated by the enzyme xanthine oxidase (XO), have been shown to be important mediators in the early pathogenesis. Ordinarily XO primarily occurs as xanthine dehydrogenase (XD) but can be converted to XO, which is the form that generates toxic oxygen metabolites. This conversion of XD to XO may take place either reversibly by way of sulfhydryl group oxidation or irreversibly by means of proteolytic cleavage of XD. This study was undertaken to investigate the mechanism of conversion of XD to XO in the FFA-, POSS-, and ISCH 2-induced pancreatitis models. CER pancreatitis was studied for comparison. After 4 hours of perfusion, pancreatitis was manifest by edema, weight gain, and hyperamylasemia in all four models. Dithiothreitol, a sulfhydryl group protector, ameliorated the weight gain in the FFA (40 +/- 14 gm to 18 +/- 13 gm; p < 0.05), POSS (28 +/- 10 gm to 9 +/- 3 gm; p < 0.05), and ISCH 2 pancreatitis (30 +/- 13 gm to 15 +/- 3 gm; p < 0.05), and ameliorated the hyperamylasemia in the POSS pancreatitis (12,062 +/- 4304 units/dl to 5877 +/- 2659 units/dl; p < 0.05). The CER pancreatitis was not ameliorated with dithiothreitol. A serine protease inhibitor of low molecular weight, phenylmethylsulfonyl fluoride, ameliorated only the CER pancreatitis (weight gain from 28 +/- 10 gm to 17 +/- 10 gm, p < 0.05; amylase activity from 38,116 +/- 6491 units/dl to 23,372 +/- 11,654 units/dl, p < 0.05), and not the FFA, POSS, or ISCH 2 pancreatitis. We conclude that in the three models of pancreatitis (FFA, POSS, and ISCH 2) that are mediated by toxic oxygen metabolites, XD is converted to XO reversibly by way of sulfhydryl group oxidation rather than irreversibly by way of proteolysis. In the CER pancreatitis, where XO does not play a role in the pathogenesis, proteolytic enzymes may be important mediators in the injury. PMID- 8417496 TI - The effect of prednisone on pancreatic islet autografts in dogs. AB - Prednisone was shown to induce hyperglycemia in dogs submitted to total pancreatectomy and pancreatic islet autotransplantation. The hyperglycemia caused by a 10-day course of prednisone, 1 mg/kg/day, starting on the day of operation was reversible within 1 week after steroid discontinuance. Three weeks after prednisone was stopped, there was no detectable adverse effect on glucose homeostasis as judged by fasting blood sugar levels and intravenous glucose tolerance test results. Four months after transplantation, glucose disappearance was delayed in animals previously treated with the prednisone compared with those previously treated with prednisone plus insulin or control animals. This was accompanied by lower insulin values on intravenous glucose tolerance testing and suggests a long-term subtle effect on islet function. The mechanism of the steroid effect is not known. However, this model could be used to test the diabetogenicity of other immunosuppressive agents including cyclosporine, FK 506, and azathioprine. PMID- 8417497 TI - Structural and functional diversity in the FGF receptor multigene family. PMID- 8417498 TI - Genomic instability and tumor progression: mechanistic considerations. PMID- 8417499 TI - Function and regulation of the human multidrug resistance gene. AB - The discovery of an energy-dependent pump system for natural product anticancer drugs has important implications for the biology of related energy-dependent transport systems as well as for the treatment of human cancer. To fully realize the therapeutic potential associated with manipulation of the multidrug transporter, it will be necessary to understand the mechanisms of action of the transporter and its mode of regulation. This review has summarized recent developments in these areas which suggest that both the activity of the pump and its genetic regulation are potential targets for new anticancer therapies. PMID- 8417500 TI - Relationship between myc oncogene activation and MHC class I expression. PMID- 8417501 TI - Lysosomes, lysosomal enzymes, and cancer. PMID- 8417502 TI - Protein tyrosine kinase growth factor receptors and their ligands in development, differentiation, and cancer. PMID- 8417503 TI - The molecular and genetic characterization of human soft tissue tumors. PMID- 8417504 TI - Selective radiofrequency catheter ablation of fast and slow pathways in 100 patients with atrioventricular nodal reentrant tachycardia. AB - One hundred patients received selective radiofrequency ablation of retrograde fast pathway (32 patients, group I) or slow pathway (68 patients, group II) to treat drug-refractory atrioventricular nodal reentrant tachycardia. In group I, a mean of 6 +/- 3 radiofrequency pulses eliminated the retrograde fast pathway. Thirty patients were free of symptoms and were not receiving antiarrhythmic drugs; two patients had accidental atrioventricular block. One patient had recurrent tachycardia and received a repeated ablation (slow pathway ablation). In group II, a mean of 9 +/- 4 radiofrequency pulses eliminated the slow pathway in 68 patients. All patients were free of symptoms and were not receiving antiarrhythmic drugs. One patient had recurrent tachycardia and received a repeated ablation. Serial follow-up electrophysiologic studies (immediate [20 to 30 minutes], early [5 to 7 days], and late [3 to 6 months]) showed that selective ablation of retrograde fast pathway was associated with nonspecific injury on the antegrade fast pathway (increase of AH interval) without effects on the slow pathway. Selective ablation of slow pathway was associated with nonspecific injury on the retrograde fast pathway in 15 patients (22%), but the antegrade fast pathway conduction parameters did not change significantly. Thus retrograde and antegrade fast pathway may be anatomically similar or have different sensitivities to radiofrequency energy, and slow pathway may be anatomically distinct from fast pathway. We conclude that (1) selective radiofrequency ablation of retrograde fast or slow pathway could cure atrioventricular nodal reentrant tachycardia with a high success rate (98%) and a low recurrence rate (2%) during a follow-up period of 6 to 18 months, but fast pathway ablation was associated with accidental atrioventricular block (5%), and (2) serial follow-up electrophysiologic studies elucidated the possible mechanisms of cure in atrioventricular nodal reentrant tachycardia. PMID- 8417505 TI - Five-year follow-up of 589 patients treated with amiodarone. AB - Between 1977 and 1986, 589 patients (age, 57 +/- 13 years; 464 men and 125 women) received amiodarone for ventricular fibrillation (VF; 147 patients), sustained (VT-S; 242 patients) or nonsustained (VT-NS; 80 patients) ventricular tachycardia, or supraventricular tachycardia (SVT; 120 patients). Mean left ventricular ejection fraction was 36 +/- 17%, with 23% in New York Heart Association functional class I, 49% in class II, 25% in class III, and 3% in class IV. Sixty-two percent had ischemic heart disease. Follow-up was 32 +/- 27 months (mean +/- SD). Life table analysis revealed that patients with VF, VT-S, and VT-NS had a cumulative incidence of sudden death of 9% at 1 year, increasing by about 3% per year. By years 2 and 5, the cumulative incidence of sudden death, VF, or VT-S recurrence was 26% and 38% and the percent of patients still taking amiodarone was 54% and 32%. For patients with SVT at years 2 and 5, the cumulative incidence of sudden death was 1% and 3%, and of sudden death or SVT recurrence the cumulative incidence was 20% and 29%. The percent of patients still taking amiodarone was 67% and 43%. Of 14 clinical variables assessed, New York Heart Association functional class was the best predictor of sudden death and arrhythmic failure and no other variable added independent predictive power. Older age and lower left ventricular ejection fraction were independent predictors of drug failure (sudden death or arrhythmic failure or need to discontinue amiodarone because of side effects). We conclude that despite its side effect profile, amiodarone is an effective and reasonably well-tolerated antiarrhythmic drug. PMID- 8417506 TI - The effects of calcium channel antagonist treatment and oxygen radical scavenging on infarct size and the no-reflow phenomenon in reperfused hearts. AB - Calcium antagonists reduce ischemic injury, and anti-free-radical interventions may reduce reperfusion injury. However, the effects of treatment with both interventions have never been investigated. In the present study, anesthetized rabbits underwent 30 minutes of coronary artery ligation, which was followed by 5.5 hours of reflow. Eight animals in each group received: (1) the calcium antagonist gallopamil during ischemia, (2) the oxygen radical scavenger superoxide dismutase during reperfusion, (3) combined treatment with gallopamil and superoxide dismutase, and (4) saline solution. All groups were similar with respect to collateral flow during ischemia and extent of risk region. Infarct size averaged 60.2% +/- 5.5% of risk region in controls and was significantly smaller (p < 0.001) in rabbits that were treated with either gallopamil (28.1% +/ 3.4%) of superoxide dismutase (29.3% +/- 3.2%). Little further reduction in infarct size was observed with combination therapy (22.9% +/- 3.2% of risk region; p = NS). Superoxide dismutase had no effects on hemodynamics, whereas gallopamil significantly reduced heart rate, mean arterial pressure, and rate pressure product. However, the reduction in infarct size that was observed in gallopamil-treated rabbits significantly exceeded the expected value in this group after corrections were made for changes in these determinants of ischemic injury. Therefore we investigated whether other factors may have contributed to the beneficial effects of gallopamil. In vitro the drug had no oxygen radical scavenging activity, nor did it exert antioxidant effects. In addition, gallopamil did not affect neutrophil function. In conclusion, in this acute model myocardial cell necrosis was significantly reduced either by administration of a calcium antagonist during ischemia or by removing oxygen radicals during reperfusion. However, superoxide dismutase administration did not further reduce infarct size when given to animals that had been treated with gallopamil. Since gallopamil has no direct effect on several mechanisms of reperfusion injury, these data suggest that calcium antagonists, by decreasing myocardial oxygen demand during ischemia, may indirectly reduce oxygen radical damage during subsequent reperfusion. PMID- 8417507 TI - Prognosis after syncope: impact of left ventricular function. AB - A cardiac cause of syncope has been associated with increased sudden death risk, whereas unexplained syncope has a benign prognosis. However, in patients who have depressed left ventricular function, the accuracy of diagnostic tests and the efficacy of therapy, such as antiarrhythmic drugs, are reduced. Previous studies of patients with syncope have not evaluated the contribution of left ventricular performance in risk stratification for sudden death. The purpose of our study of a large population of patients with syncope was to determine the impact of left ventricular dysfunction on sudden death risk if syncope is caused by a cardiac cause or remains unexplained after electrophysiologic testing. We retrospectively evaluated the relationship of left ventricular ejection fraction to sudden death prognosis in 88 consecutive patients referred for electrophysiologic testing to determine a cause of syncope. The mean age was 57 +/- 18 years, left ventricular ejection fraction was 0.41 +/- 0.20, and 66 patients (75%) had structural heart disease. In 49 patients (56%) a cardiac cause of syncope was diagnosed, and in 39 patients (44%) the cause of syncope remained unexplained after evaluation. Cardiac syncope was attributed to ventricular tachycardia in 27 patients, bradyarrhythmia in 11 patients, and supraventricular tachyarrhythmia in 11 patients. By logistic regression only structural heart disease was independently associated with cardiac cause of syncope (p = 0.003). After a mean follow-up of 790 +/- 688 days, nine patients had died suddenly, eight (89%) of whom had left ventricular ejection fraction less than 0.30.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417508 TI - Percutaneous balloon mitral valvuloplasty for mitral stenosis with and without associated aortic regurgitation. AB - Between November 1985 and December 1991, percutaneous balloon mitral valvuloplasty (PBMV) with the Inoue balloon catheter (Toray Marketing & Sales [America], Inc., New York, N.Y.) was performed in 53 patients with rheumatic mitral stenosis and associated mild to moderate aortic regurgitation. Mean left atrial pressure was 22.5 +/- 8.6 mm Hg and 9.7 +/- 5.5 mm Hg before and after PBMV, respectively (p < 0.001). The mean diastolic mitral gradient as determined by the catheter method decreased from 18.7 +/- 11.4 mm Hg to 2.1 +/- 3.1 mm Hg (p < 0.001). The echocardiographic mitral valve area was 1.0 +/- 0.2 cm2, 2.0 +/- 0.6 cm2, and 1.9 +/- 0.5 cm2, before and after PBMV and at follow-up (p < 0.001 before PBMV vs after PBMV and at follow-up). The mean diastolic mitral gradient as determined by two-dimensional and Doppler echocardiography was 19.3 +/- 8.4 mm Hg, 5.2 +/- 4.1 mm Hg, and 6.6 +/- 3.3 mm Hg, before and after PBMV and at follow up, respectively (p < 0.001). The phonocardiographic interval between the Q wave and the mitral component of the first heart sound was 85.2 +/- 15.2 msec, 74.2 +/ 13.4 msec, and 72.3 +/- 15.7 msec before and after PBMV and at follow-up (p < 0.001 before PBMV vs after PBMV and at follow-up). The phonocardiographic interval between the aortic second sound and opening snap was 73.4 +/- 18.1 msec, 88.7 +/- 9.6 msec, and 92.1 +/- 11.7 msec before and after PBMV and at follow-up (p < 0.001 before PBMV vs after PBMV and at follow-up). The voltage of P loop in the frontal plane of the vectorcardiogram was 0.25 +/- 0.04 mV, 0.21 +/- 0.04 mV, and 0.20 +/- 0.03 mV before and after PBMV and at follow-up (p < 0.001 before PBMV vs after PBMV and at follow-up). The New York Heart Association classification improved from class II in 26 patients and class III in 27 patients before PBMV to class I in 48 patients and class II in five patients after PBMV. These hemodynamic, noninvasive, and clinical results were not significantly different from those that were obtained in 112 patients with mitral stenosis without associated aortic regurgitation, who were studied during the same period in our cardiac catheterization laboratory. It was concluded that patients with rheumatic mitral stenosis are suitable candidates for PBMV whether or not they have associated aortic regurgitation of mild to moderate degree. PMID- 8417509 TI - Exercise hemodynamics in small (< or = 21 mm) aortic valve prostheses assessed by Doppler echocardiography. AB - Exercise Doppler echocardiography was used to assess hemodynamics in 25 patients with a < or = 21 mm aortic valve prosthesis (14 with a Medtronic-Hall 21 mm valve, three with a Medtronic-Hall 20 mm valve, three with a Sorin 21 mm valve, one with a Duromedics 21 mm valve, and four with a Carpentier-Edwards 21 mm valve). A symptom-limited upright bicycle exercise test was performed, and Doppler gradients were recorded during exercise. Gradients increased with exercise from 30 +/- 8/16 +/- 4 mm Hg (peak/mean) at rest to 46 +/- 12/24 +/- 7 mm Hg during exercise; both p < 0.001. Mean exercise gradient exceeded 30 mm Hg in five patients, and the highest mean gradient recorded was 37 mm Hg. Within the group of mechanical valves, gradients at exercise were similar for different types of valves. A linear relationship was found between gradients at rest and during exercise (peak r = 0.75, mean r = 0.77; both p < 0.001). Additional findings were midventricular velocities exceeding 1.5 m/sec in late systole in 10 patients (40%) and intraventricular flow (> or = 0.2 m/sec) toward the apex during isovolumic relaxation in 11 patients (44%). The patients with these velocity patterns had significantly smaller left ventricular cavities (end diastolic diameter 39.8 +/- 4.8 vs 46.5 +/- 4.2 mm, p < 0.01; end-systolic diameter 24.2 +/- 3.0 vs 28.5 +/- 4.5 mm, p = 0.013).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417510 TI - Pulmonary artery hypertension in severe aortic stenosis: incidence and mechanism. AB - We investigated the incidence and mechanism of pulmonary artery hypertension (PAH) in a consecutive series of patients with aortic stenosis who were undergoing diagnostic cardiac catheterization. Forty-five patients with severe aortic stenosis were divided into two groups: group 1 comprised 13 patients (29%) with PAH (pulmonary artery systolic pressure > 50 mm Hg); group 2 comprised 32 patients (71%) without PAH. Group 1 patients had a higher incidence of congestive heart failure, a lower left ventricular ejection fraction and cardiac index, and more mitral regurgitation as compared with group 2 patients. Of the 13 group 1 patients, 8 had a transpulmonary gradient (pulmonary artery mean pressure- pulmonary capillary wedge pressure) > or = 10 mm Hg, consistent with reactive PAH. We conclude that PAH frequently accompanies aortic stenosis and is often reactive. PMID- 8417511 TI - Usefulness of left ventricular wall stress at rest and after exercise for outcome prediction in asymptomatic aortic regurgitation. AB - Sensitive indexes for detection of left ventricular (LV) systolic performance are necessary for optimal clinical management of asymptomatic patients with aortic regurgitation (AR). To investigate the prognostic value of noninvasively determined baseline LV wall stress, we studied 10 asymptomatic patients with AR who had normal LV systolic function on two-dimensional directed M-mode echocardiography at rest and after maximal treadmill exercise. At follow-up (mean 3.6 years) three patients (group A) had progressed to decompensated LV volume overload or death related to aortic valve disease (one cardiac death and two aortic valve replacements), and seven patients (group B) remained unchanged clinically and on serial echocardiographic study. Although baseline LV chamber dimensions and systolic performance at rest were similar in the two groups of patients, LV fractional shortening after exercise and LV wall stress at rest and after exercise were significantly different (p = 0.02). Noninvasively determined baseline LV wall stress at rest and after exercise may be useful indexes for determining prognosis in asymptomatic AR. PMID- 8417512 TI - A randomized comparison between the hemodynamic effects of hydralazine and nitroglycerin alone and in combination at rest and during isometric exercise in patients with chronic mitral regurgitation. AB - Both arteriolar vasodilation with hydralazine and venodilation with nitroglycerin have been shown to favorably alter the hemodynamic profile in patients with chronic mitral regurgitation. Since these therapeutic modalities exert their effects by different mechanisms, this study was designed to assess hemodynamic response to both drugs given individually and in combination at rest and during isometric exercise in patients with chronic mitral regurgitation of various etiologies. When used individually, hydralazine caused larger increase in heart rate, cardiac index, stroke volume index, and systemic vascular resistance both at rest and during exercise. In contrast, nitroglycerin administration was associated with a superior effect on mean pulmonary artery pressure and V wave amplitude at rest and on mean right atrial and mean pulmonary artery wedge pressures at rest and during isometric exercise. In comparison with hydralazine alone, combination therapy enhanced the reduction in right atrial pressure (4 +/- 1 versus 0 +/- 2 mm Hg, p < 0.05), mean pulmonary arterial pressure (11 +/- 5 versus 1 +/- 3 mm Hg, p < 0.05), and the V wave amplitude (15 +/- 6 versus -5 +/- 3 mm Hg, p < 0.05) at rest and resulted in a larger reduction in exercise values of mean pulmonary artery pressure (-11 +/- 7 versus -3 +/- 5 mm Hg, p < 0.05) and mean pulmonary arterial wedge pressure (-11 +/- 5 versus -5 +/- 3 mm Hg, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417513 TI - Relation of exercise Doppler left ventricular filling to thallium lung uptake. AB - The exercise-induced changes in left ventricular filling in patients with coronary artery disease are poorly understood. Therefore these changes were studied in relation to a noninvasive indicator of exercise pulmonary venous congestion, the lung-to-heart (L:H) ratio on symptom-limited thallium stress testing. Fifty-six patients undergoing diagnostic treadmill testing were studied; 50 of them had technically adequate Doppler recordings and became the subjects of this study. Doppler left ventricular filling was assessed with patients in the supine position both before and after exercise. Measurements included early (E) and late (A) filling velocities, their ratio, the diastolic time-velocity integral, and the diastolic filling time. The L:H ratio was considered abnormal if it was greater than the upper 95% confidence limit for a separate group of normal subjects. Twelve subjects had a documented prior myocardial infarction, 16 had stress-induced ischemia, and 20 had abnormal L:H ratios. A greater E and a longer diastolic filling time in the group with an abnormal L:H ratio were the only postexercise measurements that differed; however, E was the only filling parameter that both differed between groups after exercise (abnormal L:H group 87 +/- 25 cm/sec; normal 68 +/- 20 cm/sec; p < 0.01) and whose change from rest to after exercise was significantly different (p < 0.05). Since Doppler velocities are directly related to instantaneous gradients, the higher E in patients with evidence of exercise pulmonary congestion suggests a higher exercise early diastolic left atrial pressure. PMID- 8417514 TI - Assessment of left ventricular diastolic function in dilated cardiomyopathy with cine magnetic resonance imaging: effect of an angiotensin converting enzyme inhibitor, benazepril. AB - The effects of angiotensin converting-enzyme inhibitor, benazepril, on diastolic function in patients with dilated cardiomyopathy, with (n = 4) or without (n = 11) mitral regurgitation, were examined with the time-volume curve of the left ventricle derived from cine magnetic resonance images. Peak filling rate/end systolic volume and ejection fraction were increased in the group without regurgitation (p < 0.01) but not in the group with regurgitation after treatment. There was a strong correlation between peak filling rate/end-systolic volume and ejection fraction (r = 0.89) and between the change in peak filling rate/end systolic volume and that in ejection fraction after treatment (r = 0.74) in the group without regurgitation. These findings suggest that in some patients with dilated cardiomyopathy benazepril has favorable effects on diastolic function, which seem to be related to improvement in systolic function. This drug may not be as beneficial in patients with dilated cardiomyopathy complicated by mitral regurgitation. PMID- 8417515 TI - Noninvasive diagnosis of coarctation of the aorta in the presence of a patent ductus arteriosus. AB - Although the diagnosis of coarctation of the aorta in the full-term neonate is straightforward when no ductus arteriosus is present, identification of coarctation of the aorta when a ductus arteriosus is patent can be difficult. A patent ductus arteriosus is frequently found in association with coarctation of the aorta, and it may remain open for many days. Thus a reliable method to rapidly identify coarctation of the aorta would obviate the need for cardiac catheterization or an in-hospital observation period until the ductus arteriosus closes spontaneously. Echocardiographic/Doppler examination of 19 consecutive full-term neonates with a birth weight of > 2.9 kg, who subsequently underwent surgical repair of coarctation of the aorta, were reviewed. Fourteen patients with similar weights, a patent ductus arteriosus, and a normal aortic arch were matched for comparison. Based on the findings, the following diagnostic criteria for coarctation of the aorta are suggested: isthmic diameter less than or equal to 3 mm or isthmus equal to 4 mm together with the Doppler finding of continuous antegrade flow in the isthmal segment. Coarctation of the aorta in the full-term neonate can be ruled out when the flow within the ductus arteriosus is exclusively from aorta to pulmonary artery or when the isthmic diameter is equal to or greater than 5 mm. PMID- 8417516 TI - The cough test is superior to the Valsalva maneuver in the delineation of right to-left shunting through a patent foramen ovale during contrast transesophageal echocardiography. AB - A patent foramen ovale may result in paradoxical embolization and serious morbidity. Thus a sensitive method to diagnose a patent foramen ovale is important. It is unknown whether the cough test or the Valsalva maneuver is superior in delineating right-to-left shunting through a patent foramen ovale during contrast transesophageal echocardiography. Thus we studied 73 consecutive patients (53 men and 20 women), aged 54 +/- 16 years (range 18 to 79 years), during elective transesophageal echocardiography. Contrast transesophageal echocardiography was performed from a four-chamber view during quiet respirations, Valsalva maneuver, and cough test. In the entire group the incidence of a patent foramen ovale was higher during the cough test (32/73) as compared with the Valsalva maneuver (24/73, p < 0.025) and quiet respirations (18/73, p < 0.005). All subjects with a patent foramen ovale during the Valsalva maneuver had a positive contrast transesophageal echocardiogram during the cough test. In subjects (n = 55) without a patent foramen ovale during quiet respirations, the incidence of a patent foramen ovale was higher during the cough test (15/55) as compared with the Valsalva maneuver (9/55, p < 0.05). In a subgroup (N = 17) of patients with nonhemorrhagic stroke (n = 11), transient ischemic attack (n = 2), or peripheral embolus (n = 4), the cough test had a higher yield (9/17) in delineating a patent foramen ovale as compared with the Valsalva maneuver (7/17) but did not reach statistical significance. These data demonstrate that the cough test is superior to the Valsalva maneuver in delineating a patent foramen ovale during contrast transesophageal echocardiography.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417517 TI - Risk of endocarditis in transesophageal echocardiography. AB - The risk of endocarditis associated with transesophageal echocardiography was studied in 101 patients. To evaluate possible bacteremia, blood cultures were performed on samples from consecutive patients who did not have clinical or laboratory evidence of infection. The broth blood culture Signal system was used in all patients, and additionally, the lysis-centrifugation technique was performed in a subgroup of 40 patients to further ameliorate recovery of rapidly phagocytosed germs. Comprehensive criteria for differentiation between true bacteremia and possible contamination were provided by means of simultaneous blood sampling from two separate venipuncture sites and skin specimens from the venipuncture area. Oropharyngeal specimens were cultured for evaluation of possible association of oropharyngeal flora with positive blood culture findings. They revealed facultative pathogenic isolates, as well as physiologic residental flora, in 15 patients. All blood isolates that were recovered simultaneously 6 minutes after the procedure were found to be sterile. Correspondingly, clinical follow-up for 2 weeks was uneventful with regard to episodes of infection. These results indicate that the risk of bacteremia associated with transesophageal echocardiography is extremely low. Thus endocarditis prophylaxis is not required for this procedure. PMID- 8417518 TI - Cardiac and skeletal muscle disease in systemic sclerosis (scleroderma): a high risk association. AB - To examine the possible relationship between cardiac and skeletal muscle disease in systemic sclerosis, we reviewed computerized records of 1095 consecutive patients with systemic sclerosis. One hundred eighty three (17%) had skeletal myopathy. Thirty-nine (21%) of the 183 fulfilled criteria for myocardial disease, compared with 90 (10%) of the 912 without myopathy (p < 0.0001.) Nineteen (10%) of the 183 had clinical CHF compared with 38 (4%) of the remainder (p < 0.002.) Fifteen (8%) of the patients with myopathy died of cardiac causes compared with 27 (3%) of the 912 without myopathy (p < 0.002.) Twenty-five patients with coexistent myopathy and myocardial disease, in the absence of other identifiable contributing causes, were identified. This group was characterized by a high incidence of cardiac conduction abnormalities (60%) and by the severity of the myocardial dysfunction and arrhythmias, both atrial and ventricular that they experienced. Eighteen of these 25 patients died; 12 (67%) died suddenly. Eight of the 18 (44%) had intractable CHF, which directly contributed to their deaths. Myocardial fibrosis was the predominant histologic abnormality at autopsy. However, autopsy of a patient who died in the context of acute "myocarditis" showed severe myocytolysis with contraction band necrosis but without inflammation or fibrosis; this is consistent with possible ischemically mediated injury. We conclude that skeletal and cardiac muscle disease in systemic sclerosis are associated. Patients with myopathy are at increased risk for CHF, sustained symptomatic arrhythmias, and cardiac death, particularly sudden death. PMID- 8417519 TI - Heart rate variability and baroreflex sensitivity in myocardial infarction. PMID- 8417520 TI - Advantages and limitations of methods to detect, localize, and quantitate intracardiac right-to-left and bidirectional shunting. PMID- 8417521 TI - Identification of high-risk patients with left main and three-vessel coronary artery disease using stepwise discriminant analysis of clinical, exercise, and tomographic thallium data. AB - This large-scale study examined the ability of stepwise discriminant analysis of clinical, exercise, and thallium tomographic data to detect high-risk patients with three-vessel or left main disease. There were 834 patients, 229 with three vessel or left main disease (group 1) and 605 (group 2) with either two-vessel disease (n = 236), one-vessel disease (n = 195), or no coronary artery disease (n = 174). The two groups were different in age, exercise heart rate, ST segment depression during exercise, exercise systolic blood pressure, abnormal thallium scans, reversible perfusion defects, extent of thallium abnormality, number of vascular territories with perfusion abnormalities, left ventricular cavity dilatation, and increased lung thallium uptake. On multivariate stepwise discriminant analysis, only three variables were independent predictors of high risk. These included multivessel thallium abnormality (F = 107, p < 0.001), exercise heart rate (F = 27, p < 0.001), and ST segment depression (F = 8, p < 0.01). Based on these three variables, patients could be stratified into three categories with different prevalences of left main or three-vessel disease; the prevalence was 53% in 239 patients, 24% in 271 patients, and 12% in 324 patients. Thus high-risk patients with left main or three-vessel disease can be identified by exercise thallium tomographic imaging that uses a model based on stepwise discriminant analysis. The thallium data are far more powerful than the clinical or treadmill exercise data. PMID- 8417522 TI - Thermal-perfusion balloon coronary angioplasty: in vivo evaluation. AB - The goal of this study was to develop and test a new radio frequency thermal balloon system to allow longer balloon inflations at lower temperature levels than have been used with standard (laser) thermal balloon angioplasty. Radio frequency thermal capabilities were combined with perfusion balloon technology, creating a thermal-perfusion balloon catheter system for prolonged thermal inflations. Twenty-five dogs underwent thermal-perfusion angioplasty at 37 degrees, 60 degrees, or 80 degrees C for 1.5, 5, or 15 minutes with angiographic and morphologic assessments at 24 hours (n = 17) or 4-6 weeks later (n = 8). Treated segments and side branches remained patent. No coronary spasm, occlusive thrombus, or ischemic myocardial infarction occurred. Histologic extent of thermal injury in treated segments was proportional to treatment duration. Thus the thermal-perfusion balloon angioplasty system may be safely applied in canine coronary arteries. Integrating thermal and perfusion technologies provides prolonged treatment duration at moderate temperatures without excessive tissue damage. PMID- 8417523 TI - Electrophysiologic recovery in postischemic, stunned myocardium despite persistent systolic dysfunction. AB - Previous investigators have hypothesized that myocardial "stunning" may result either from a primary impairment in excitation or from electromechanical dissociation. Thrombolytic therapy and angioplasty have increased the importance of understanding the electrophysiologic effects of brief ischemia followed by reperfusion. We investigated the electrophysiologic properties of mechanically dysfunctional stunned myocardium in 18 dogs anesthetized with pentobarbital (30 mg/kg, intravenously administered). After thoracotomy, the proximal anterior descending coronary artery was occluded for 15 minutes, which was followed by 20 minutes of reperfusion. At baseline, peak ischemia, and 20 minutes of reperfusion, local electrogram durations, activation times, and refractory periods were measured from 12 standardized sites within the ischemic and border zones. Echocardiographic percentage of systolic wall thickening confirmed normal preischemic and markedly reduced postischemic function in the investigated region. Despite the marked electrophysiologic abnormalities observed in the ischemic zone during ischemia, mean electrogram duration, calculated conduction velocity, and mean effective refractory period after 20 minutes of reperfusion had returned almost to baseline values 39.2 +/- 11.5 msec versus 37.2 +/- 12.1 msec, 0.65 +/- 0.15 m/sec versus 0.68 +/- 0.15 m/sec, and 134 +/- 14 msec versus 131 +/- 8 msec, respectively. Corresponding mean values within the ischemic border zone were similarly close to baseline values after reperfusion. There was no significant difference in local heterogeneity (coefficient of variation) within the ischemic or border zone after reperfusion versus baseline values. Although the postischemic electrophysiologic status returned to normal, systolic thinning and dyskinesis persisted in the region of measurement. The contractile dysfunction that results from reperfusion-induced injury can thus occur in the setting of apparent excitation-contraction uncoupling. PMID- 8417524 TI - Recurrent angina due to a left internal mammary artery-to-pulmonary artery fistula. PMID- 8417525 TI - Left main coronary artery compression in patent ductus arteriosus. PMID- 8417526 TI - Collateral circulation provided by an accessory coronary artery arising from the noncoronary sinus of Valsalva in obstructive coronary arterial disease. PMID- 8417527 TI - Localized right atrial tamponade and right-to-left shunting as a complication of pericarditis after myocardial infarction. PMID- 8417528 TI - Exercise-induced ST-segment depression: relationship to nonocclusive coronary artery spasm. PMID- 8417529 TI - Recurrent vasospastic angina of more than thirteen years' duration. PMID- 8417530 TI - Myocardial infarction after amphetamine use. PMID- 8417531 TI - Late coronary aneurysm formation after directional atherectomy. PMID- 8417532 TI - Change of the isoform of creatine phosphokinase MM in hypertrophic cardiomyopathy. PMID- 8417533 TI - Reversible obstructive hypertrophic cardiomyopathy after dexamethasone therapy for bronchopulmonary dysplasia. PMID- 8417534 TI - Successful radiofrequency ablation of both atrioventricular nodal reentrant and circus movement tachycardia. PMID- 8417535 TI - Low-frequency intracardiac ultrasonographic imaging before and after balloon pulmonary valvuloplasty. PMID- 8417536 TI - Diagnosis of acute intraoperative pulmonary thromboembolism by transesophageal echocardiography. PMID- 8417537 TI - Simultaneous measurement of pulmonary artery diastolic pressure by Doppler echocardiography and catheterization in patients with patent ductus arteriosus. PMID- 8417538 TI - Mitral and aortic paravalvular leaks with hemolytic anemia. PMID- 8417539 TI - Percutaneous transcatheter pericardial interventions: aspiration, biopsy, and pericardioplasty. PMID- 8417540 TI - Myocardial hibernation in the infarcted region cannot be assessed from the presence of stress-induced ischemia: usefulness of delayed image of exercise thallium-201 scintigraphy. AB - To examine the relationship between the improvement of wall motion in infarcted regions after percutaneous transluminal coronary angioplasty (PTCA) and thallium 201 uptake in the delayed image of exercise thallium-201 scintigraphy before PTCA, 14 patients with anterior old myocardial infarction were studied. Exercise thallium-201 scintigraphy was performed before PTCA of left anterior descending artery, and mean percent thallium-201 uptake of abnormal segments was calculated in the initial and 4-hour delayed images. Left ventricular angiography was performed during catheterization, before, and 4 to 13 months after PTCA; and regional ejection fraction of anterior wall was calculated. Atrial pacing stress test with the measurement of lactate concentration of aorta and great cardiac vein was performed during catheterization before PTCA. In five patients with mean percent thallium-201 uptake in the delayed image < or = 50% (group I), regional ejection fraction did not increase after PTCA (23% +/- 9% to 24% +/- 12%). In the other nine patients with mean percent thallium-201 uptake > 50% (group II), regional ejection fraction increased significantly after PTCA (39% +/- 18% to 47% +/- 14%; p < 0.05). There was no significant difference in regional ejection fraction, lactate extraction ratio during maximal pacing, and the redistribution of exercise thallium-201 scintigraphy between the two groups before PTCA. Thus the delayed image before PTCA is useful to detect reversible nonfunctioning viable myocardium (hibernating myocardium) in the infarcted region. However, the wall-motion abnormality and the degree of stress-induced ischemia in the infarcted region before PTCA may not be necessarily useful for the detection of hibernating myocardium. PMID- 8417541 TI - Significance of ST-segment elevation during ambulatory monitoring after acute myocardial infarction. AB - The significance of ST segment elevation during ambulatory monitoring after acute myocardial infarction was examined in 203 patients. Ambulatory monitoring was performed both early (mean 6.4 days [range 3 to 15]; N = 201) and late (38 days [range 22 to 93]; N = 177), and 174 patients underwent exercise treadmill testing (38 days [range 22 to 93]). Cardiac events (death, reinfarction, and coronary revascularization) were documented during a 1-year follow-up period. ST elevation (all silent) occurred in 25 of 201 patients (12%) on early monitoring but in only 4 of 177 (2%) on late monitoring (p < 0.001). Compared with patients (N = 148) without any ST deviation, those with early ST elevation had more pericarditis (8/25 [32%] vs 23/148 [16%]; p = 0.089) but no more angina or exercise ischemia. The mortality rate tended to be higher in patients with early ST elevation (4/25 [16%] vs 10/148 [7%]; p = 0.24), but ST elevation was too infrequent to be a valuable prognostic indicator. ST elevation is not uncommon during ambulatory monitoring early after myocardial infarction but is rare during later monitoring. Such ST elevation is almost always silent, does not usually reflect myocardial ischemia, and is not a useful prognostic indicator. PMID- 8417542 TI - The prognostic value, clinical, and angiographic characteristics of patients with early postinfarction angina after a first myocardial infarction. AB - To evaluate the prognostic value of early postinfarction angina and its relationship to clinical and angiographic variables, 231 consecutive patients who had a first myocardial infarction were studied. All underwent cardiac catheterization within 10 weeks after admission to the hospital. There were no differences in basic characteristics or in ejection fraction, extent or severity of coronary artery disease, or collateral circulation between the patients with early postinfarction angina (n = 27) and those without early postinfarction angina (n = 204) except for the incidence of angina before myocardial infarction. Patients with early postinfarction angina had exercise-induced angina (42% vs 21%; p < 0.025) more frequently and shorter exercise duration (6.9 +/- 2.5 minutes vs 8.3 +/- 2.5 minutes; p = 0.007). Early postinfarction angina was associated with a significantly higher event rate (15% vs 4%; p < 0.025) and a significantly higher mortality rate (15% vs 3%; p < 0.005) in the first year after infarction but not during the subsequent 4-year follow-up. In patients with early postinfarction angina, stress test results had no predictive value for future cardiac events in contrast to patients without early postinfarction angina in whom ST-segment depression as observed on the stress test ECG and exercise duration had predictive value for future cardiac events. In patients with early postinfarction angina there was no relationship between the incidence of events and the number of diseased vessels in contrast to patients without early postinfarction angina who had a high incidence of events when three-vessel disease was present (16% vs 62%; p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417543 TI - Progressive decrease in myocardial ischemia assessed by intracoronary electrocardiogram during successive and prolonged coronary occlusions in angioplasty. AB - Progressive decrease in chest pain and surface ECG changes are commonly observed during successive balloon inflations in coronary angioplasty, which suggests a decrease in myocardial ischemic response. To assess this hypothesis, we continuously recorded intracoronary ECGs during four balloon inflations; each of the inflations was maintained to a minimum of 120 seconds in 19 patients who had significant stenosis in the left anterior descending artery and normal left ventricular function. Three successive QRS-T complexes were analyzed on surface and intracoronary ECGs for measurements of ST-segment elevation 60 milliseconds after the J point. Surface ECG changes were compared with intracoronary ECG changes. On intracoronary ECG, ST area (in square millimeters) and T wave amplitude (in millimeters) were also computed. Chest pain was noted as present or absent during each successive balloon inflation. Ability of intracoronary ECG to detect myocardial ischemia, which was defined as ST-segment elevation greater than 1 mm during balloon inflations 1 to 4, was 89%, 89%, 84%, and 74%, respectively and was higher than that of surface ECG, which was 68%, 63%, 68%, and 58%, respectively. On intracoronary ECG, when compared with the first balloon inflation, a significantly smaller increase in ST-segment elevation was noted during each subsequent balloon inflation, whereas a significantly smaller increase in ST area and T wave amplitude was noted only during balloon inflation 4. The number of patients who experienced chest pain decreased from 15 to 13, 10 and 6 from the first to the fourth balloon inflation. This report demonstrates a progressive decrease in myocardial ischemic response during successive and prolonged balloon occlusions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417544 TI - Progressive coronary dilation: an angioplasty technique that creates controlled arterial injury and reduces complications. AB - The incidence of dissection or acute closure during coronary angioplasty has remained unchanged in spite of increased operator experience and technologic advances. To test the hypothesis that progressive coronary dilation, that is, predilation of the stenosis with a smaller balloon (2.0 or 2.5 mm) and then maximal dilation with an optimally sized (3.0, 3.25, 3.5, 3.75, or 4.0 mm) balloon may produce controlled injury and thus reduce the incidence of major complications, the procedural success rate and acute complications of progressive coronary dilation were analyzed in 1087 patients (1486 vessels) and compared with other large series. To determine whether progressive coronary dilation would improve success rates for complex lesions, the last 167 vessels were also prospectively characterized by American College of Cardiology/American Heart Association criteria. Of the 1248 vessels with partial occlusions, the success rate was 98.7%. Attempts to dilate total occlusions in 16% (353) of vessels yielded a success rate of 88%. The primary success rates for types A, B, and C lesions were 100%, 97%, and 91%, respectively. Multivessel angioplasty with progressive coronary dilation was done in 32.4% of cases. Acute closure, major dissection, emergency coronary bypass, periprocedural myocardial infarction, and in-hospital death were noted in 1.4%, 1.3%, 0.7%, 0.8%, and 0.09% of the patients, respectively; the incidence was significantly lower than in previously reported series. Mean residual stenosis was 20.0% +/- 10.6%. Thus progressive coronary dilation by controlled injury to the plaque offers a high primary success rate; low residual stenosis; and markedly lower incidence of acute closure, major dissection, emergency coronary bypass, and death in dilation of both simple and complex lesions. PMID- 8417545 TI - Ionic contrast agent-mediated endothelial injury causes increased platelet deposition to vascular surfaces. AB - Contrast agent-mediated endothelial injury may be clinically relevant to the development of acute thrombosis after coronary interventions. We sought to investigate the extent to which contrast agents increase platelet deposition by measuring deposition of indium-111 radiolabeled platelets in an isolated perfused rabbit carotid artery model. Carotid artery segments were perfused at physiologic temperature, pressure, and shear. Vessels were subjected to angioplasty or no angioplasty before exposure to either buffer, diatrizoate (high osmolal/ionic), ioxaglate (low osmolal/ionic), or ioversol (low osmolal/nonionic). Subsequent deposition of indium-111 radiolabeled platelets was quantified. In vessels without balloon angioplasty, platelet deposition (platelets/cm2) was 110,000 +/- 95,000 for buffer perfused vessels, 280,000 +/- 210,000 for vessels perfused with diatrizoate, 290,000 +/- 160,000 for vessels perfused with ioxaglate, and 130,000 +/- 98,000 for vessels perfused with ioversol. After balloon angioplasty, platelet deposition was 1,300,000 +/- 590,000 for buffer controls, 1,800,000 +/- 320,000 for diatrizoate-perfused vessels, 1,500,000 +/- 450,000 for ioxaglate perfused vessels, and 1,000,000 +/- 180,000 for ioversol-perfused vessels. In vessels without balloon angioplasty, diatrizoate and ioxaglate increased platelet deposition 2.5-fold and 2.6-fold, respectively, relative to buffer-perfused vessels (p < 0.05 and p < 0.01), whereas no increase was seen with ioversol. After balloon angioplasty, diatrizoate increased platelet deposition 1.4-fold over control (p < 0.05), whereas ioxaglate and ioversol showed no statistically significant increase. We conclude that ionic contrast media may cause more endothelial injury and associated localized platelet deposition than nonionic contrast media. These findings may be relevant to coronary interventions, specifically with regard to acute closure and chronic restenosis. PMID- 8417546 TI - Aspirin inhibits platelet activity but does not attenuate experimental atherosclerosis. AB - We evaluated the antiatherosclerotic potential of aspirin, a platelet inhibitor, in lipid-fed rabbits (0.3% cholesterol diet). Seventy-five male New Zealand white rabbits were divided into treated or control groups. The treated groups were given aspirin by daily gavage for 12 weeks (1 mg/kg, 10 mg/kg, 30 mg/kg, and 60 mg/kg) and 10 rabbits served as controls. Increased bleeding time was observed in the aspirin-treated groups (average, 58 +/- 10 seconds to 75 +/- 17 seconds; p < 0.001). Only high-dose aspirin (60 mg/kg/day) significantly inhibited platelet aggregation (1.04 +/- 0.15 vs 0.67 +/- 0.14; p < 0.05). Seventeen additional rabbits had aortic endothelial injury produced by a balloon catheter. Eight of them were given aspirin (40 mg/kg/day), and the other nine served as controls. The average percent of surface lesions and lesion thickness of the aorta and pulmonary artery were not significantly reduced by aspirin. These results show that at doses that cause antiplatelet effects, aspirin does not attenuate atherosclerosis. PMID- 8417547 TI - Coronary arterial ectasia: increased prevalence in patients with abdominal aortic aneurysm as compared to occlusive atherosclerotic peripheral vascular disease. AB - Coronary artery ectasia (CAE) is the saccular or fusiform dilatation of a coronary artery. CAE is found in 1.2% to 4.9% of patients at autopsy or during angiographic studies, with a similar prevalence of CAE found in patients with atherosclerotic peripheral vascular disease (PVD). Abdominal aortic aneurysm (AAA) and CAE are similar in pathogenesis and histology. To determine whether CAE occurs more frequently in patients with AAA than in occlusive forms of atherosclerotic PVD, a review of coronary angiograms was performed in patients who underwent cardiac catheterization and vascular reconstruction for AAA or occlusive atherosclerotic PVD of the lower extremities. Of 72 patients with AAA, 15 had CAE (20.8%) compared with only 2 of 69 patients with atherosclerotic PVD (2.9%) (p < 0.003). CAE was predominantly discrete, located in the left coronary system, and associated with significant coronary atherosclerosis. CAE may be more prevalent in patients with AAA resulting from a similar pathogenetic process. PMID- 8417548 TI - Coronary flow reserve in patients with dilated cardiomyopathy. AB - Coronary flow reserve was studied in patients with dilated cardiomyopathy. A 3F coronary Doppler catheter was placed in the proximal left anterior descending artery in each of 10 patients with dilated cardiomyopathy (CDM group), seven patients with coronary artery disease that involved only the left anterior descending artery (CAD group), and seven patients with chest pain syndrome and normal hearts (control group). Coronary flow reserve was calculated as the ratio of the maximum mean coronary blood flow velocity after intracoronary administration of papaverine (10 mg) to resting flow velocity (M/R). The time until maximum flow velocity was reached after papaverine administration (Tmax) was also measured. M/R was lower in the DCM (p < 0.001) and CAD (p < 0.001) groups when compared with the control group. Tmax was not abnormal in the DCM group but was prolonged in the CAD group (p < 0.05). In the DCM group, the M/R ratio correlated with the left ventricular end-diastolic pressure (r = -0.69; p < 0.05), the left ventricular end-diastolic volume index (r = -0.7; p < 0.05), the ejection fraction (r = 0.82; p < 0.01), the left ventricular mass (r = -0.7; p < 0.05), and the left ventricular end-diastolic wall stress (r = -0.84; p < 0.001). These results indicate that coronary flow reserve was decreased in patients with dilated cardiomyopathy and that the mechanism of its reduction may differ from that in patients with coronary artery disease. PMID- 8417549 TI - Transthoracic defibrillation: effect of dual-pathway sequential pulse shocks and single-pathway biphasic pulse shocks in a canine model. AB - To determine whether dual-pathway sequential shocks and single-pathway biphasic shocks improved the efficacy of transthoracic defibrillation, we delivered single or sequential truncated waveform shocks of variable duration, voltage, and direction (polarity) to three groups of closed-chest dogs. Dual-pathway sequential shocks were assessed in group 1 (eight animals), biphasic shocks with a single pathway were compared in 11 dogs (group 2), and the effect of varying the duration of the biphasic shocks was assessed in group 3 (four animals). There was no improvement in success rates of the intervention shocks compared with a standard single "control" shock at any energy level. In this experimental model unidirectional or biphasic sequential shocks given over single or dual pathways were not superior to standard single-pulse transthoracic defibrillation. PMID- 8417550 TI - Suicidal behavior and risk factors among runaway youths. AB - OBJECTIVE: The goal of this study was to describe suicide attempts and risk factors among runaway adolescents. METHOD: A structured interview format was used to assess suicidal behavior and suicide-related risk factors among a consecutive series of 576 predominantly black or Hispanic runaway adolescents at intake into four publicly funded runaway programs in New York City over a 2-year period. There were no significant differences in age, gender, race/ethnicity, education, or socioeconomic status among the adolescents at the four runaway program sites. RESULTS: Thirty-seven percent of the youths had previously attempted suicide, and 44% of the attempters had made an attempt within the previous month. Females were significantly more likely than males to have attempted suicide and to be depressed. Male runaways were far more likely to have attempted suicide than nonrunaway male adolescents described in previously published reports. Runaways with histories of attempting suicide were significantly more likely to be currently suicidal and depressed. CONCLUSIONS: This study indicates the need for systematic screening of runaway adolescents for suicidal ideation at residential shelters for youths. PMID- 8417551 TI - Suicidal behaviors in adult psychiatric outpatients, I: Description and prevalence. AB - OBJECTIVE: Because the findings of previous studies of suicidal behaviors in psychiatric outpatients may not necessarily generalize to outpatients with a wide spectrum of psychiatric diagnoses, the authors evaluated the prevalence of suicidal behaviors in a large general psychiatric outpatient clinic whose patients represented a full spectrum of psychiatric illness. METHOD: A total of 651 patients participated in the study between 1987 and 1989. These patients had sought treatment at the outpatient psychiatry department of a private nonprofit hospital. Before being interviewed for treatment, all patients were given a comprehensive self-rating survey packet that included the Harkavy Asnis Suicide Survey and the Hopkins Symptom Checklist-90. The Harkavy Asnis Suicide Survey is a self-report questionnaire that assesses demographic variables, current and past history of suicidal behaviors of the patient as well as family members and peers, and a detailed description of each previous attempt. RESULTS: Fifty-five percent of the patients had a history of suicidal ideation, and 25% reported at least one previous suicide attempt. Approximately half of the suicide attempters reported multiple attempts. The predominant methods of attempt were overdose (53%), jumping (17%), and wrist cutting (17%). Suicidal behavior was prevalent in most diagnostic groups. The rates of suicidal ideation among patients with mood disorders (major depression, dysthymia, and bipolar disorder), adjustment disorders, and alcohol/substance abuse were significantly greater than that of the patients with generalized anxiety disorder. CONCLUSIONS: The authors conclude that suicidal behavior is prevalent among patients who seek treatment in a general outpatient department. PMID- 8417552 TI - Dawn simulation treatment of winter depression: a controlled study. AB - OBJECTIVE: This study sought to determine whether dawn simulation was superior to a shorter dimmer "placebo" dawn signal in treating winter depression. METHOD: In a randomized, parallel design, 22 patients with winter depression were treated with either 1 week of a 2-hour dawn simulation peaking at 250 lux or 1 week of a 30-minute dawn simulation peaking at 0.2 lux. The subjects were told that they would receive either a "gradual" dawn or a "rapid" dawn reaching an intensity that would be dimmer than standard bright light treatment. At the end of both the baseline week and the treatment week, subjects were assessed in a blind manner with the Hamilton Rating Scale for Depression. Analysis of covariance was used to compare the two dawn treatments. RESULTS: The 2-hour, 250-lux dawn simulation resulted in Hamilton depression scale scores that were significantly lower than scores after the 30-minute, 0.2-lux dawn simulation. CONCLUSIONS: This study indicates that dawn simulation is an effective treatment for winter depression. PMID- 8417553 TI - A double-blind crossover trial of imipramine and phenelzine for outpatients with treatment-refractory depression. AB - OBJECTIVE: Data from controlled studies concerning the response rates of patients to a second antidepressant medication after they have been unresponsive to a systematic trial of another antidepressant are extremely useful to clinicians for rational prescription of pharmacotherapy. Such information allows making an accurate prognosis, sustaining realistic hope in the patient, and achieving the best possible therapeutic outcome. This study was designed to add to the scanty literature available on this subject. METHOD: Eighty-nine mood-reactive, nonmelancholic, mainly chronically depressed outpatients at a university research clinic who were unresponsive to vigorous double-blind trials of imipramine or phenelzine were crossed over to treatment with the other drug under double-blind conditions. RESULTS: Of 46 patients previously unresponsive to imipramine who completed phenelzine treatment, 31 (67%) responded to phenelzine. Of 22 patients previously unresponsive to phenelzine who completed imipramine treatment, nine (41%) responded to imipramine. The difference in response rates was statistically significant. Even after they had shown no response to 7 weeks of placebo and 6 weeks of imipramine treatment, 10 (83%) of 12 patients who then completed treatment with phenelzine responded. CONCLUSIONS: These data suggest that among chronically ill, mood-reactive depressed patients with many symptoms of atypical depression, phenelzine is strikingly effective in those who have been nonresponders to imipramine and should be tried in such patients. PMID- 8417554 TI - Association of depression with 10-year poststroke mortality. AB - OBJECTIVE: Depression has been linked to higher than expected mortality from natural causes, particularly among elderly patients with physical illness. The authors examined the effect of depression on mortality among a group of stroke patients followed up for 10 years. METHOD: A consecutive series of 103 patients was assessed for major or dysthymic (minor) depression approximately 2 weeks after stroke with the use of a structured mental status examination and DSM-III diagnostic criteria. Vital status was determined for 91 of these patients 10 years later. RESULTS: Forty-eight (53%) of the 91 patients had died. Patients with diagnoses of either major or minor depression were 3.4 times more likely to have died during the follow-up period than were nondepressed patients, and this relationship was independent of other measured risk factors such as age, sex, social class, type of stroke, lesion location, and level of social functioning. The mortality rate among depressed patients with few social contacts was especially high: over 90% had died. CONCLUSIONS: These results indicate that depressed mood following stroke is associated with an increased risk of subsequent mortality. Patients who are depressed and socially isolated seem to be particularly vulnerable. PMID- 8417555 TI - Intelligence and brain structure in normal individuals. AB - OBJECTIVE: This study was designed to evaluate the relation between intelligence and a variety of measures of brain structure. METHOD: Magnetic resonance imaging scans were used to measure the volume of the intracranial cavity, cerebral hemispheres, lateral ventricles, temporal lobes, hippocampus, caudate, and cerebellum, as well as the overall volume of gray matter, white matter, and CSF, in 67 healthy, normal volunteers. Intelligence was measured with the Wechsler Adult Intelligence Scale--Revised. RESULTS: Full-scale IQ was found to be significantly correlated with intracranial, cerebral, temporal lobe, hippocampal, and cerebellar volume but not with caudate and lateral ventricle volume. There were also significant correlations of full-scale, verbal, and performance IQ with overall gray matter volume but not with white matter or CSF volume. Gender differences were noted in the pattern and number of correlations between the volume of the brain and its subregions and full-scale, verbal, and performance IQ. CONCLUSIONS: The results suggest that the size of some cerebral structures may account for a significant, but modest, proportion of the variance in human intelligence. PMID- 8417556 TI - Trends in research in two general psychiatric journals in 1969-1990: research on research. AB - OBJECTIVE: The authors describe the characteristics of psychiatric research over the past two decades as captured in the articles published by two general psychiatric journals. METHOD: A total of 1,236 articles were drawn from The American Journal of Psychiatry and Archives of General Psychiatry for October through September 1969-1970, 1979-1980, and 1989-1990. Articles were assigned to one of five categories. Research articles were then further categorized as to methodological approach and field of research as well as specific topic areas. Funding sources listed for each research article were also indexed. RESULTS: Over time and in both journals, the percentage and number of research articles have risen, with a concomitant reduction in case reports, opinion papers, and "other" articles. Categories of research design were fairly consistent across time and in both journals. Percentages of articles on specific fields and topics indicated an increasing emphasis on biological studies, especially those in clinical psychobiology, as well as a sharp move away from general categories to a more disorder-specific orientation. Reporting of funding sources has substantially increased. CONCLUSIONS: The large proportion of research articles published in these two important general psychiatric journals reflects editorial policies, changing audience expectations, and the availability of new research tools. Systematic analysis of trends in psychiatric research and other forms of research on research can be useful approaches to assessing the growth and utilization of knowledge in the field, to planning how to most effectively use limited research resources, and to increasing public support for research. PMID- 8417557 TI - Physician-assisted suicide: the dangers of legalization. AB - The authors examine physician-assisted suicide in the light of what is known about suicide and terminal illness, exploring the potential for abuse if legalization occurs. The elderly, those frightened by illness, and the depressed of all ages would be potential victims. The authors discuss the cases that have received public attention as illustrative of these abuses. PMID- 8417558 TI - Magnetic resonance imaging of schizophrenia-like psychoses associated with cerebral trauma: clinicopathological correlates. AB - Three patients with schizophrenia-like psychosis and two with schizoaffective like psychosis who experienced cerebral trauma before the onset of their illness underwent clinical and magnetic resonance imaging evaluation. Each patient with a schizophrenia-like psychosis, but neither of those with a schizoaffective-like psychosis, showed abnormalities confined to or including the left temporal lobe. These observations complement recent findings in schizophrenia. PMID- 8417559 TI - CSF homovanillic acid in schizotypal personality disorder. AB - CSF concentrations of homovanillic acid (HVA) were measured in 10 patients with schizotypal personality disorder and 14 patients with other personality disorders. The patients with schizotypal personality disorder had higher CSF HVA concentrations than the patients with other personality disorders. Furthermore, the psychotic-like schizotypal symptoms correlated positively with the CSF HVA concentrations. These results suggest a central dopaminergic dysfunction associated with the psychotic-like symptoms of schizotypal personality disorder. PMID- 8417560 TI - Impaired eye tracking in undergraduates with schizotypal personality disorder. AB - Qualitative ratings of smooth pursuit eye movements were significantly worse for 14 undergraduates with DSM-III-R schizotypal personality disorder than for 18 comparison subjects. The groups did not differ on IQ, indicating that deficits in smooth pursuit eye movements in schizotypal personality disorder are not a function of cognitive deficits. PMID- 8417561 TI - Transient compulsive foraging behavior associated with crack cocaine use. AB - Compulsive foraging behavior associated with use of crack cocaine involves compulsively searching the environment for possibly misplaced pieces of crack. Of 41 crack cocaine addicts evaluated, 33 (80.5%) reported at least some compulsive foraging associated with use of crack; 21 (51.2%) reported such behavior as always associated with crack use. The mean length of time spent in compulsive foraging was 90 minutes. Cocaine-induced foraging may represent a drug-induced model of a type of compulsive behavior. PMID- 8417562 TI - Antidepressant withdrawal: prospective findings. PMID- 8417563 TI - Tardive dyskinesia-like syndromes with clomipramine. PMID- 8417564 TI - Huntington's disease and propranolol. PMID- 8417565 TI - Euphoria with buspirone after fluoxetine treatment. PMID- 8417566 TI - Fluoxetine and prolonged erection. PMID- 8417567 TI - Treatment of panic disorder in coronary artery disease. PMID- 8417568 TI - Carbamazepine and plasma levels of clozapine. PMID- 8417569 TI - Anorexia nervosa and lower vulnerability to infections. PMID- 8417570 TI - Violent antisocial behavior and Wisconsin Card Sorting Test performance in cocaine addicts. PMID- 8417571 TI - Dissociation and eating. PMID- 8417572 TI - Folie a deux involving a dog. PMID- 8417573 TI - Isolated but serious acts of self-mutilation. PMID- 8417574 TI - Antisemitism and the collective unconscious. PMID- 8417575 TI - Efficacy of brief psychotherapy. PMID- 8417576 TI - The phenomenological and conceptual interface between borderline personality disorder and PTSD. AB - OBJECTIVE: The authors explore the conceptual and phenomenological interface between posttraumatic stress disorder (PTSD) and borderline personality disorder as well as the therapeutic and research implications of this interface. METHOD: They systematically review the relevant empirical, conceptual, and clinical literature. RESULTS: These seemingly separate disorders are related. Borderline personality disorder is often shaped in part by trauma, and individuals with borderline disorder are therefore vulnerable to developing PTSD. CONCLUSIONS: The authors draw a distinction between the enduring effects that traumas can have on formation (or change) of axis II personality traits (including those found in borderline personality disorder) and acute symptomatic reactions to trauma, called PTSD, that are accompanied by specific psychophysiological correlates. They describe the implications of these conclusions for DSM-IV, therapy, and future research. PMID- 8417577 TI - Network therapy for addiction: a model for office practice. AB - Individual therapists in office practice are often considered to have limited effectiveness in treating alcohol and drug dependence. In this article the author describes network therapy, an approach developed to assure greater success in such treatment. It uses psychodynamic and behavioral therapy while engaging the patient in a support network composed of family members and peers. A cognitive behavioral model of addiction, based on the role of conditioned withdrawal in relapse, is described. Related techniques for securing abstinence are then reviewed; they augment individual psychotherapy to help patients avoid relapse caused by the affective and environmental cues that precipitate drug seeking. The role of social cohesiveness as a vehicle for engaging patients in treatment is outlined next, along with a related technique for enhancing an addicted patient's commitment to the therapy. This is done by using the patient's family and peers as a therapeutic network to join the patient at intervals in therapy sessions. The network is managed by the therapist to provide cohesiveness and support, undermine denial, and promote compliance with treatment. The author presents applications of the network technique designed to sustain abstinence and describes means of stabilizing members' involvement. Applications of network therapy to ambulatory detoxification, disulfiram and naltrexone administration, relapse prevention, and contingency contracting are reviewed. PMID- 8417578 TI - Comparison of cognitive-behavioral and supportive-expressive therapy for bulimia nervosa. AB - OBJECTIVE: The authors compared the effectiveness of 4 months (18 sessions) of cognitive-behavioral and supportive-expressive therapy for bulimia. METHOD: Sixty patients obtained from clinical referrals to an eating disorders program who met modified DSM-III-R criteria for bulimia nervosa were randomly assigned to the two conditions. Treatments were delivered in an individual format, on an outpatient basis, by experienced therapists using treatment manuals. The primary outcome measures were self-induced vomiting, binge eating, and attitudes toward body weight and shape, which were assessed by self-report and structured interview. RESULTS: Fifty patients completed treatment, 25 in each condition. Both treatments led to significant improvements in specific eating disorder symptoms and in psychosocial disturbances. Supportive-expressive therapy was just as effective as cognitive-behavioral therapy in reducing binge eating. Where treatment differences were found, they favored cognitive-behavioral therapy. Cognitive-behavioral therapy was marginally superior in reducing the frequency of self-induced vomiting; 36% of the patients who received cognitive-behavioral therapy and 12% of those who received supportive-expressive therapy abstained from vomiting in the last month of treatment. Cognitive-behavioral therapy was significantly more effective in ameloriating disturbed attitudes toward eating and weight, depression, poor self-esteem, general psychological distress, and certain personality traits. CONCLUSIONS: These results moderately favor cognitive behavioral therapy over supportive-expressive therapy for bulimia nervosa, but follow-up is required to determine the durability of outcome with both modalities. The findings must be interpreted with caution since the selected clinical sample in this study may not represent the bulimia nervosa population. PMID- 8417579 TI - HIV seroprevalence among homeless patients admitted to a psychiatric inpatient unit. AB - OBJECTIVE: This study was conducted to determine the seroprevalence of HIV-1 antibodies among hospitalized homeless mentally ill patients. METHOD: From December 1989 through May 1991 the authors collected discard blood samples from patients consecutively admitted to a psychiatric unit designated for the care of severely mentally ill persons removed from the streets of New York City. The blood samples were tested for HIV-1 antibodies, and the results were analyzed for associations with age, gender, ethnicity, male homosexual activity, and use of injected drugs. RESULTS: The HIV seroprevalence was 6.4% (13 of 203 samples). Patients between ages 18 and 39 accounted for 51.2% of the admissions and 84.6% of the 13 positive results, a seroprevalence of 10.6% for this subsample. Patients under age 40 were more than six times as likely to test positive for HIV antibodies as those 40 or over. Ethnicity did not predict seropositivity. Women were as likely as men to be infected. Although clinicians had noted high-risk behavior on the charts for only three (23.1%) of the 13 positive cases, a recorded history of use of injected drugs was associated with a 6.5-fold greater risk of HIV seropositivity. CONCLUSIONS: One in every 16 patients admitted to the special unit was HIV positive. Age under 40 and use of injected drugs were strongly associated with seropositivity. Because information on high-risk behavior was infrequent, the reasons for younger patients' greater risk are unclear. The homeless mentally ill require outreach efforts to reduce the risk of acquiring or transmitting HIV. PMID- 8417580 TI - Antisocial personality disorder and HIV infection among intravenous drug abusers. AB - OBJECTIVE: Antisocial personality disorder in drug abusers has been associated with poor treatment outcome. The authors examined the relationship between diagnosis of antisocial personality and HIV infection. METHOD: Subjects were 272 intravenous drug abusers, 140 (52%) of whom were in methadone treatment. Subjects were given an HIV risk behavior interview before diagnostic interviewing and HIV testing. RESULTS: Using the DSM-III-R definition, the authors found that 119 (44%) of the subjects met criteria for antisocial personality. Significantly more of the subjects with antisocial personality (18% [N = 21] than of the subjects without antisocial personality (8% [N = 12]) had HIV infection. The diagnosis of antisocial personality disorder was associated with a significantly higher odds ratio of infection independent of ethnicity, gender, and treatment status. CONCLUSIONS: Antisocial personality is a risk factor for HIV infection among intravenous drug abusers. PMID- 8417581 TI - Structural abnormalities in deficit and nondeficit schizophrenia. AB - OBJECTIVE: Previous studies have suggested the involvement of the frontal and parietal cortices and thalamus in a neural circuit underlying the production of primary enduring negative or deficit symptoms of schizophrenia. The purpose of this study was to examine whether structural changes in the proposed circuit are associated with the production of deficit symptoms. METHOD: Magnetic resonance imaging was used to measure the volume of selected circuit brain regions (i.e., the prefrontal region and caudate) and noncircuit brain regions (i.e., the amygdala/hippocampus complex) in 17 deficit and 24 nondeficit schizophrenic outpatients and 30 normal comparison subjects. RESULTS: Right and left total prefrontal volumes discriminated deficit from nondeficit patients, with prefrontal volumes being smaller in nondeficit patients. There were no differences between the two schizophrenic subgroups in left caudate or right and left amygdala/hippocampus complex volumes. The right caudate was larger in deficit patients, but the difference between the two schizophrenic subgroups was not significant. There were no differences between deficit and normal subjects on any prefrontal region measure. Nondeficit patients had smaller total right and left prefrontal volumes than normal subjects. Both schizophrenic subgroups had larger left caudate volumes and smaller right and left amygdala/hippocampus complex volumes than the normal subjects. There was a trend for deficit patients to have larger right caudate volumes. CONCLUSIONS: These results suggest that structural changes in the prefrontal region are not responsible for deficit symptoms. The caudate, particularly the right caudate, may be associated with the production of these symptoms. PMID- 8417582 TI - Psychiatric illnesses in families of subjects with schizophrenia-spectrum personality disorders: high morbidity risks for unspecified functional psychoses and schizophrenia. AB - OBJECTIVE: The authors determined morbidity risks for psychiatric illnesses in the families of probands with schizophrenia-spectrum personality disorders. METHOD: Subjects were recruited from the community through newspaper advertisements. Subjects were identified as having schizophrenia-spectrum personality disorders (N = 30) if they met at least three, four, or three DSM-III R criteria for schizoid (N = 14), schizotypal (N = 20), and/or paranoid (N = 15) personality disorder, respectively. The comparison subjects had no psychiatric diagnoses (N = 8) or had other personality disorders (N = 12); none of the subjects in either group had any DSM-III-R axis I diagnosis. Trained interviewers collected family history information about the relatives of the two groups; the interviewers were blind to the probands' diagnoses. RESULTS: The risks for schizophrenia, other functional psychoses, and schizophrenia-spectrum personality disorders were significantly higher in the relatives of subjects with schizophrenia-spectrum personality disorders than in the families of the comparison subjects. CONCLUSIONS: The high rate of schizophrenia in the families of probands with schizophrenia-spectrum personality disorders is consistent with the previous findings of higher than normal rates of these personality disorders in the biological relatives of schizophrenic patients. The significance of the high rate of unspecified functional psychoses is unclear. Use of the family study method, by which valid differential diagnosis of psychoses is possible, is indicated. The results from the current study do not rule out the possibility that the schizophrenia-spectrum personality disorders are related to psychoses in general rather than specifically to schizophrenia. PMID- 8417583 TI - Psychiatric care and health insurance reform. AB - Concerns about cost, access, and quality of health care in the United States have led to a variety of legislative proposals that would reform our health care system and its financing. Health insurance benefits for mental illness, including substance abuse, are treated differently from medical/surgical benefits, with stricter limits on outpatient visits and hospital days. Medicare, Medicaid, and most private health insurance plans contain this historic disparity of coverage for mental illness compared to general medical illness. Psychiatric services are also distinguishable because of the large public sector reimbursement for mental illness treatment and support. Principles for a more equitable design of mental health benefits include a non-discriminatory approach; payment on the basis of service rather than diagnosis; application of cost containment for care of mental illness on the same basis as care of general medical illness; retention of the public sector as a backup system for high-cost, long-term care; encouragement of lower-cost alternatives to the hospital through the development of a continuum of care; and a recognition of the distinction between psychotherapy and medical management. All current approaches to universal health care fall short of these principles. A research agenda is needed now more than ever in order to articulate the case for complete coverage of mental illness and substance abuse. PMID- 8417584 TI - Length of stay and recidivism in schizophrenia: a study of public psychiatric hospital patients. AB - OBJECTIVE: Psychiatric beds in public hospitals have decreased 80% since 1955, but admissions have risen correspondingly, largely because of high recidivism rates. Decreases in numbers of beds have been partly achieved by shortening the length of stay, which lessened by half between 1970 and 1980. This study was undertaken to determine whether duration of hospital treatment affects the rate and rapidity of relapse among schizophrenic patients. METHOD: Data on 1,500 patients from 10 state hospitals were gathered for 18 months after initial discharge. Predictor variables included age, sex, marital status, race, number of previous admissions, location of the facility, and length of stay. Data were analyzed by survival analysis with a Cox regression model for two times to initial relapse: 30 days and 18 months (outcome). RESULTS: Length of stay was significantly related to each time to relapse after the effects of number of previous admissions and age were partialed out. Facility location was not predictive, but intrahospital effects were tested by examining the data on the largest facility; again, length of stay significantly predicted relapse. CONCLUSIONS: Although the magnitude of the effect was small, the clinical significance of the findings is the greater likelihood that brief-stay patients will be rehospitalized within 30 days after discharge than will patients treated for longer periods. Brief hospitalization seems generally applicable to psychiatric populations, but there may be a small but important group of seriously mentally ill patients for whom other alternatives are possibly more appropriate and should be explored. PMID- 8417585 TI - Relative performance of for-profit psychiatric hospitals in investor-owned systems and nonprofit psychiatric hospitals. AB - OBJECTIVE: The authors analyzed the differences in operational and financial performance between 42 matched pairs of for-profit psychiatric hospitals belonging to multifacility organizations and nonprofit psychiatric hospitals for the fiscal years ending in 1986 through 1990. METHOD: The pairs of short-term hospitals were matched according to location, standard metropolitan statistical area, or wage index. Analyses were based on data on these hospitals from the Health Care Financing Administration. The groups of variables studied included the hospitals' operational performance and productivity, profitability and payer mix, revenue and expenses, and capital structure. Differences in the mean values of the variables for the for-profit hospitals and the nonprofit hospitals were analyzed by pairwise t tests. RESULTS: The for-profit organization hospitals had significantly higher net revenue, lower salary expenses, and higher profits than the nonprofit hospitals. Patients in the for-profit hospitals had longer stays, and these hospitals had fewer full-time employees per adjusted inpatient day and per adjusted discharge. CONCLUSIONS: The higher prices and operating margins of the for-profit hospitals belonging to investor-owned systems reflect the profit maximizing goal of these facilities. The ability of for-profit organization hospitals to achieve economies of scale in expenses, however, was not evident except in the case of salary expenses. PMID- 8417586 TI - Enhanced suppression of cortisol following dexamethasone administration in posttraumatic stress disorder. AB - OBJECTIVE: The authors investigated the possibility of enhanced negative feedback sensitivity of the hypothalamic-pituitary-adrenal (HPA) axis in posttraumatic stress disorder (PTSD) by using a low dose of dexamethasone. METHOD: Baseline blood samples were obtained at 8:00 a.m., and 0.5 mg of dexamethasone was administered to 21 male patients with PTSD and 12 normal age-comparable men at 11:00 p.m. Cortisol and dexamethasone levels were measured 9 and 17 hours after dexamethasone administration. RESULTS: After correction for differences in dexamethasone levels, the PTSD patients showed greater suppression of cortisol in response to dexamethasone than did the normal subjects. This was true even in patients meeting concurrent diagnostic criteria for major depression. CONCLUSIONS: The data support earlier studies showing that HPA abnormalities in PTSD are different from those seen in depression and suggest that the low-dose dexamethasone suppression test may be a potentially useful tool for differentiating the two syndromes and further exploring differences in their pathophysiology. PMID- 8417587 TI - Anticipatory stress in children and adolescents. AB - OBJECTIVE: Circumstances surrounding the New Madrid earthquake prediction on Dec. 3, 1990, offered a unique opportunity to study the effects of a disaster warning stage on children and adolescents. METHOD: An initial structured interview was administered to 553 third- and 10th-grade students before December 3, with follow up interviews conducted 6-8 weeks later. RESULTS: This study documents the existence of a mild but prevalent PTSD-like reaction that arose from exposure to a prediction of disaster. CONCLUSIONS: Further study of anticipatory stress reactions is needed to provide insights into the development of methods for providing support to children during disaster warnings. PMID- 8417588 TI - Obsessions in obsessive-compulsive disorder with and without Gilles de la Tourette's syndrome. AB - OBJECTIVE AND METHOD: Although obsessive-compulsive disorder commonly occurs in many patients with Gilles de la Tourette's syndrome, little is known about the obsessions and compulsions of Tourette's syndrome and whether they differ from those seen in pure obsessive-compulsive disorder. The authors prospectively studied 10 subjects with obsessive-compulsive disorder and 15 subjects with obsessive-compulsive disorder and comorbid Tourette's syndrome by using the Yale Brown Obsessive Compulsive Scale, the Leyton Obsessional Inventory, and a new questionnaire designed to emphasize the differences in symptoms between these two groups. RESULTS: Subjects with comorbid obsessive-compulsive disorder and Tourette's syndrome had significantly more violent, sexual, and symmetrical obsessions and more touching, blinking, counting, and self-damaging compulsions. The group with obsessive-compulsive disorder alone had more obsessions concerning dirt or germs and more cleaning compulsions. The subjects who had both disorders reported that their compulsions arose spontaneously, whereas the subjects with obsessive-compulsive disorder alone reported that their compulsions were frequently preceded by cognitions. CONCLUSIONS: There are phenomenologic differences between obsessive-compulsive disorder and obsessive-compulsive disorder with comorbid Tourette's syndrome that may reflect differential involvement of neurochemical and neuroanatomic pathways. PMID- 8417589 TI - Premonitory urges in Tourette's syndrome. AB - OBJECTIVE: Tourette's syndrome traditionally has been viewed as a hyperkinetic movement disorder characterized by involuntary motor and phonic tics. Many patients, however, describe their tics as a voluntary response to premonitory urges. This cross-sectional study evaluated premonitory urges and related phenomena in subjects with tic disorders. METHOD: A total of 135 subjects with tic disorders, aged 8 to 71 years, completed a questionnaire concerning their current and past tic symptoms. Subjects were asked to describe and, if possible, localize their premonitory urges. The Yale Global Tic Severity Scale was used to assess current tic severity. The method of case finding does not provide prevalence data for premonitory urges. RESULTS: Ninety-three percent of the subjects reported premonitory urges. Anatomical regions with the greatest density of urges were the palms, shoulders, midline abdomen, and throat. Eighty-four percent of the subjects reported that tics were associated with a feeling of relief. A substantial majority (92%) also indicated that their tics were either fully or partially a voluntary response to the premonitory urges. CONCLUSIONS: While epidemiological studies of tic disorders have yet to incorporate questions concerning premonitory urges, these results suggest that such urges may be commonplace in adolescent and adult subjects with tic disorders. These results challenge the conventional wisdom that tic behaviors are wholly involuntary in character. They also implicate brain regions involved in the processing of sensorimotor information in the pathobiology of tic disorders. PMID- 8417590 TI - Bone mineral density in postmenopausal women as determined by prior oral contraceptive use. AB - The long-term consequences of prior oral contraceptive use for bone mineral density were examined in 239 postmenopausal women, 35.1% of whom reported prior oral contraceptive use. Women who had used oral contraceptives for 6 or more years had significantly higher bone densities of the lumbar spine and femoral neck, but not of the ultradistal wrist or radius, than women who never used oral contraceptives. PMID- 8417591 TI - Infant feeding in Queensland, Australia: long-term trends. AB - Infant feeding practices were retrospectively ascertained in a random cohort of parous women (mean age 54.8 years) from Brisbane, Australia. Reported proportions of infants who were ever breast-fed fell from around 90% before 1960 to around 70% in the early 1970s, with some subsequent increase. Similar but stronger trends were reported in proportions of infants breast- but not bottle-fed. Few maternal characteristics were associated with feeding practices, but women with more education appear to have led both the early retreat from and the later return to breast-feeding. PMID- 8417592 TI - The socioeconomic correlates of hysterectomies in the United States. AB - The purpose of this study was to examine the relationship between incidence of prior hysterectomy and education, income, and race. Data concerning previous hysterectomy and socioeconomic information were collected from 12,465 women 18 years or older as part of the Behavioral Risk Factor Surveillance System conducted in 16 states in 1988. The results indicate that women with less education and lower incomes were more likely to have had a hysterectomy. Race was not related to hysterectomy rate. PMID- 8417593 TI - An update on New York City's dramatic increase in low birthweights. AB - Monthly time-series data on rates of low birthweight and very low birthweight in New York City from 1963 to 1990 indicate that the dramatic increase, particularly among Blacks beginning in 1984, appears to have peaked in 1988. Based on vital statistics records, the rate of low birthweight for Blacks fell from 13.1% in 1988 to 12.1% in 1990. The rate of low birthweight among Whites and Hispanics has fallen slightly since 1988. PMID- 8417594 TI - Alcohol, cigarette, and marijuana use among fourth-grade urban schoolchildren in 1988/89 and 1990/91. AB - A public-school-based epidemiological survey of fourth-grade students in Washington, DC, was performed in 1988/89 (n = 4675) and 1990/91 (n = 4678). Comparisons of data for the two periods revealed that the lifetime prevalence of self-reported alcohol use, alcohol use without parental knowledge, and smoking more than a puff of cigarettes declined; marijuana use and cigarette experimentation did not. Seven variables were associated with use. Declines were observed in perceived peer pressure to use; seeing a family member/friend selling drugs; and being offered alcohol, cigarettes, or marijuana by peers. No declines were observed in family use, perceived friends' use, being bothered a lot if best friends use, or seeing someone else selling drugs. PMID- 8417595 TI - Public housing subsidies may improve poor children's nutrition. PMID- 8417596 TI - The effect of previous cesarean sections on current cesarean rates. PMID- 8417597 TI - Italian infant mortality attributable to low birthweight. PMID- 8417598 TI - Addressing low birthweight and infant mortality. PMID- 8417599 TI - Beliefs vs behaviors in healthcare decision making. PMID- 8417600 TI - Decisions near the end of life: professional views on life-sustaining treatments. AB - OBJECTIVES: How do health care professionals assess the care of hospital patients near the end of life? Are physicians and nurses aware of and in agreement with national recommendations regarding patients' rights to forgo life-sustaining medical treatments and to receive adequate pain control? METHODS: We surveyed 687 physicians and 759 nurses in 5 hospitals. RESULTS: Almost half (47%) of all respondents and fully 70% of the house officers reported that they had acted against their conscience in providing care to the terminally ill. Four times as many respondents were concerned about the provision of overly burdensome treatment than about undertreatment. CONCLUSIONS: In summary, many physicians and nurses were disturbed by the degree to which technological solutions influence care during the final days of a terminal illness and by the undertreatment of pain. However, changes in the care of dying patients may not have kept pace with national recommendations, in part because many physicians and nurses disagreed with and may have been unaware of some key guidelines, such as the permissibility of withdrawing treatments. PMID- 8417601 TI - Commentary: balancing life and death--proceed with caution. AB - Hospital professionals' decisions to permit death are amalgams of medical, ethical, and legal judgments. Medical education and socialization and the business of health all focus on offering and providing treatment, not on facilitating death. Some patients are suspicious that rights to refuse care will foster abandonment by care providers. Lawyers and risk managers often let exaggerated fears of future liability limit patients' and families' rights. The culture of medical institutions must change to accommodate notions of negotiated death. PMID- 8417602 TI - Social differences in Swedish infant mortality by cause of death, 1983 to 1986. AB - OBJECTIVE: We sought to investigate social differences in Swedish infant mortality by cause of death. METHODS: All live single births in Sweden between 1983 and 1986 to mothers 15 to 44 years old with Nordic citizenship were studied. The causes of death were classified into six major groups. Mother's education was used as a social indicator. Logistic regression analysis was used with identical models for all groups of causes of death. RESULTS: There were 355,601 births and 2012 infant deaths. Only for sudden infant death syndrome were significant social differences found, with crude odds ratios of 2.6 for mothers with less than 10 years of education and of 1.9 for mothers with 10 to 11 years, compared with 1.0 for mothers with 15 years or more. After adjusting for age, parity, and smoking habits, these ratios were no longer significant. CONCLUSIONS: The social differences obtained could be explained by the fact that mothers with less education smoke more, are younger, and have higher parity than those with more education. PMID- 8417603 TI - A comparison of prenatal care use in the United States and Europe. AB - OBJECTIVES: We sought to describe prenatal care use in the United States and in three European countries where accessibility to prenatal care has been reported to be better than it is in the United States. METHODS: We analyzed the 1980 US National Natality Survey, the 1981 French National Natality Survey, a 1979 sample of Danish births, and a survey performed from 1979 to 1980 in one Belgian province. RESULTS: The proportion of women who began prenatal care late (after 15 weeks) is highest in the United States (21.2%) and lowest in France (4.0%). This contrasts with the median number of visits, which is greater in the United States (11) than in Denmark (10) or in France (7). Across all maternal ages, parities, and educational levels, late initiation of prenatal care is more frequent in the United States, and median number of visits in the United States is equal to or higher than that in the other countries. CONCLUSIONS: In countries that offer nearly universal access to prenatal care, women begin care earlier during pregnancy and have fewer visits than women in the United States. PMID- 8417604 TI - The health consequences of maquiladora work: women on the US-Mexican border. AB - OBJECTIVES: As more US companies take jobs to Mexico, complaints are growing that the assembly plants (maquiladoras) exert adverse effects on workers' health. This study assessed the health of female electronic and garment maquiladora workers, comparing them with women employed in services and non-wage earners. METHODS: A survey was administered to 480 women living in Tijuana in 1990. The sample was stratified by occupation and length of employment. Functional impediments, nervousness, depression, and sense of control were used as outcome variables, controlling for other confounders. RESULTS: Despite working longer hours, receiving lower wages, and having less decision latitude and education, maquiladora workers were not worse off than service workers. Maquiladora workers reported similar incidences of depression and lack of control over life. Electronics workers, especially, had lower incidences of nervousness and functional impediments, after controlling for other confounders. Also, maquiladora work did not add an extra health burden compared with non-wage earners. CONCLUSIONS: The adverse effects of maquiladoras previously reported may have been exaggerated. Subjective factors, including negative attitudes toward economic adversity and work dissatisfaction, were stronger predictors of health than were objective indicators. PMID- 8417605 TI - Cervical cancer and health care resources in Newark, New Jersey, 1970 to 1988. AB - OBJECTIVES: In the past, the predominantly Black population of Newark, NJ, had little access to programs promoting or providing Pap tests. The ratio of in situ to invasive cases of cervical carcinoma was markedly reduced in all age categories, indicating inadequate screening for this cancer in this population. Funding became available to provide and publicize Pap smears but ceased after 5.5 years. We examined the effect of these changes in funding. METHODS: Data came from all Newark hospitals and practitioners and from the state cancer registry. There are now data on incidence of in situ and invasive cervical cancer in Newark from 1970 through 1988, including years before, during, and after program funding. RESULTS: The ratio of in situ to invasive cervical cancer increased and decreased in a striking parallel with the provision and subsequent cessation of funding. CONCLUSIONS: Cessation of funding of education and screening programs can result in resumption of an unfavorable in situ/invasive cervical carcinoma ratio in a poor population. PMID- 8417606 TI - Demographic factors in the use of children's mental health services. AB - OBJECTIVES: This study was designed to (1) determine mental health service use by children of varying age, sex, socioeconomic status, and urbanicity of residence; (2) compare the prevalence of mental disorder in children in these groups; and (3) determine the extent to which differences in service use are consonant with the prevalence differences. METHODS: Data on psychiatric diagnoses and service use were taken from a random longitudinal sample of 760 children. Information was gathered by interviews of mothers and of youth aged 12 to 21. RESULTS: Significant lags in mental health service use were found for youth 18 to 21 years of age, for those living in rural and semirural areas, and for those in middle income families. To some extent, these service use differences paralleled differences in diagnostic rates. However, when diagnostic differences were controlled, the same patterns of service use inequalities were present. CONCLUSIONS: Mental health service use rates for youth vary by age, urbanicity, and family income. The underservice of middle-income and rural children may reasonably be ascribed to access problems; we explore explanations for the underservice of older youth. PMID- 8417607 TI - Demographic predictors of mammography and Pap smear screening in US women. AB - OBJECTIVES: Proven screening technologies exist for both breast and cervical cancer, but they are underused by many women. We sought to evaluate the effect of demographic characteristics on the underuse of mammography and Pap smear screening. METHODS: We analyzed responses from 12,252 women who participated in the 1987 National Health Interview Survey Cancer Control Supplement. Demographic profiles were produced to target severely underserved groups of women. RESULTS: Low income was a strong predictor of mammography underuse, as was Hispanic ethnicity and other race, low educational attainment, age greater than 65, and residence in a rural area. A strong predictor of never having had a Pap smear was never having been married; however, the importance of this characteristic is difficult to interpret in the absence of data on sexual activity. Hispanic women and women of other races of all ages and all income levels underused Pap smear screening, as did older women, particularly older Black women. CONCLUSIONS: The tendency of women to underuse screening technologies varies greatly across levels of basic demographic characteristics. The importance of these characteristics differs for mammography screening versus Pap smear screening. PMID- 8417608 TI - Towards a more inclusive model of women's health. PMID- 8417609 TI - Recognizing illicit drug use by pregnant women: reports from Oregon birth attendants. AB - OBJECTIVES: This study was undertaken to determine the prevalence of recognized prenatal illicit substance abuse and the characteristics of women being identified as illicit drug users in a statewide population-based cohort. METHODS: During a 1-month period, birth attendants of all singleton births in Oregon (n = 3200) were surveyed regarding their knowledge of prenatal illicit drug use by women who gave birth. Birth certificates were linked to surveys after removal of personal identifiers. RESULTS: Illicit drug use was recognized in 5.2% of delivering women. Nearly half had used cocaine, methamphetamine, or heroin. Recognized users were significantly more likely than nonusers to be unwed and to have used tobacco or alcohol, have received inadequate prenatal care, and have public assistance as a source of payment. Drug use was recognized twice as frequently by practitioners who routinely questioned their patients about it compared with those who relied on clinical judgment or the occurrence of complications during pregnancy. Birth certificate reporting identified only 41% of recognized users. CONCLUSIONS: Oregon practitioners are identifying seven times as many drug-using women as can be accommodated by available treatment programs for this population. Increased efforts are needed to ensure the adequacy of resources necessary to cope with the problem as already recognized. PMID- 8417610 TI - AIDS-related knowledge, perceptions, and behaviors among impoverished minority women. AB - OBJECTIVES: The aims of this study were to (1) describe AIDS-related knowledge, perceptions, and risky behaviors of impoverished African-American and high- and low-acculturated Latina women; (2) delineate relationships involving high-risk behaviors; and (3) determine whether risky behaviors differ by race and levels of acculturation. METHODS: Survey instruments were administered to 1173 impoverished women of color residing in homeless shelters and drug recovery programs. RESULTS: Differences based on ethnicity and level of acculturation were found in AIDS related knowledge, perceived risk of acquiring AIDS, and risky behaviors. Low acculturated Latinas reported low perceived risk and were least likely to engage in illegal drug use and sexual activity with multiple partners. Intravenous drug use was most prevalent among high-acculturated Latinas, whereas nonintravenous drug use and high-risk sexual activity was most prevalent among African-American women. CONCLUSIONS: The data indicate the need for culturally sensitive AIDS prevention programs for women that deal with general issues of drug use and unprotected sex, and that include separate sessions for women of different ethnic backgrounds and acculturation levels to address specialized areas of concern. PMID- 8417611 TI - Survival with AIDS in Massachusetts, 1979 to 1989. AB - OBJECTIVES: The goal of the study was to determine survival time after diagnosis of acquired immunodeficiency syndrome (AIDS) and to identify predictors of survival. METHODS: We conducted a population-based prospective survival analysis of all Massachusetts-resident adult AIDS patients diagnosed from January 1, 1979, through December 31, 1988. RESULTS: Median survival was 406 days, with a 5-year survival rate of 3%. Age older than 40 years (P = .001), a diagnosis other than Kaposi's sarcoma (P = .001), and a history of intravenous drug use (P < or = .01) were associated with shorter survival after confounding was controlled. Survival increased as year of diagnosis became more recent (P < .0001). This temporal effect was strongest for patients with Pneumocystis carinii pneumonia. Individuals with Kaposi's sarcoma, Hispanics, homosexual men who were concurrent intravenous drug users, and residents of the greater Boston standard metropolitan statistical area, excluding the city of Boston, did not experience increases in survival over time. CONCLUSIONS: With the exception of cases initially defined by Kaposi's sarcoma, recently diagnosed AIDS case subjects survive longer than those diagnosed earlier in the epidemic. Further work is needed to determine whether this effect is due to lead-time bias or better treatment after diagnosis. PMID- 8417612 TI - Risk factors associated with HIV infection among male prostitutes. AB - OBJECTIVES: This study documents the human immunodeficiency virus (HIV) and sexually transmitted disease seroprevalence rate for male prostitutes, identifies the risk factors for HIV, and provides baseline information for the development and implementation of appropriate prevention and intervention strategies. METHODS: Structured interviews were conducted with and blood samples were collected from 235 actively working male prostitutes in Atlanta, Georgia, from July 1988 through July 1991. RESULTS: The HIV seroprevalence was 29.4%; 25.1% of the sample had seromarker for syphilis and 58.3% for hepatitis B. Multivariate logistic regression analysis showed the following significant HIV risk factors: history of receptive anal sex with nonpaying partners, serologic history of hepatitis B or syphilis, and history of childhood physical abuse. CONCLUSIONS: The reported seroprevalence rates among these male prostitutes indicate they are a high-risk group. The striking difference in HIV seroprevalence by sexual orientation may warrant special attention. Considering the public health consequences, there is a clear need for innovative HIV prevention and intervention among these men. PMID- 8417613 TI - Residential releases of number 2 fuel oil: a contributor to indoor air pollution. AB - OBJECTIVES: Analysis of data from the New York City Fire Department showed that residential fuel oil releases frequently occur in quantities ranging from 5 to 1000 gal, primarily from storage tank leaks and overfill. A risk assessment was conducted to determine whether Number 2 fuel oil basement spills pose a significant risk to human health. METHODS: Exposure was derived from a simulated field study spill of Number 2 fuel oil in a townhouse basement to develop emission rates for the indicator constituent xylene. Distribution of xylene throughout the townhouse was determined using a multizone contaminant dispersal model. RESULTS: Spills of 85 and 21 gal resulted in xylene exposure estimates as high as 20 and 5 mg/kg/day, respectively. CONCLUSIONS: A spill of about 21 gal or more of Number 2 fuel oil would present a human health risk for central nervous and reproductive systems for 8 days or longer. Tank inspection and supervised delivery would provide effective prevention at minimal expense. PMID- 8417614 TI - Building-related asthma in Denver office workers. AB - OBJECTIVES: Reported cases of building-related asthma and hypersensitivity pneumonitis among workers in a Denver office building prompted an epidemiologic investigation. METHODS: A cross-sectional, self-administered survey of employees in the office building of the reported cases was compared with that of employees in a comparison building. RESULTS: A significant excess of respiratory disease existed among 512 Denver workers (case building) compared with 281 office workers in a suburban agency (control building). Denver employees had a higher prevalence of respiratory symptoms, and the prevalence of physician-diagnosed asthma with onset or exacerbation since building occupancy was 4.9 times greater for Denver employees than for suburban employees. Asthma was not associated with any particular ventilation system in the building. Water incursion from a below-grade wall may have contributed to the problem, but the etiology is unknown. CONCLUSIONS: This investigation provided evidence of office building-related asthma. Individual cases may be sentinel events for other cases of work-related asthma or hypersensitivity pneumonitis and may indicate a need for public health investigation of remediable causes. PMID- 8417615 TI - Class, race, and infant mortality in the United States. AB - As a result of Sweden's efforts to eliminate poverty and to provide comprehensive health care, there are only small social class differences in infant mortality. The wider social differences in US infant mortality are a consequence of less consistent and thorough attempts at social equity and universal health care. US Black infant mortality continues to be twice that of Whites, and the excess may partially result from racism. Public health research should examine the role of racism in infant mortality and develop interventions to eliminate racism and its effects on the health of Black Americans. PMID- 8417616 TI - Telephone health surveys: potential bias from noncompletion. AB - OBJECTIVES: Little is known about the effect of noncompletion on telephone surveys of health issues. This paper identifies a little-studied source of noncompletion, passive refusal, and evaluates its contribution to noncompletion bias along with two other sources: noncooperation and noncontact. Passive refusals include respondents who repeatedly request callbacks and households where interviewers repeatedly encounter an answering machine. METHODS: Measures of noncompletion (noncooperation, passive refusal, and noncontact), demographic and socioeconomic characteristics, health risk factors, and indicators of health care access and health status were collected through the Orange County Health Surveys on 4893 respondents. The surveys sampled by random-digit dialing and interviewed by computer-assisted telephone. RESULTS: Passive refusals have a substantial impact on completion rates and bias due to noncompletion. Commonly used definitions for completion rates may underestimate the bias due to noncompletion because they omit passive refusals. After we controlled for demographic and socioeconomic factors, few noncompletion biases appeared on selected health indicators. CONCLUSIONS: These results suggest improved reporting of completion rates and support a multivariate framework for studying noncompletion in telephone health surveys. PMID- 8417617 TI - Regulation of the 12-O-tetradecanoyl-phorbol-13-acetate-induced inhibition of intercellular communication. AB - Effects of the tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and protein kinase C (PKC) inhibitors on cell-cell communication were studied in a normal rat liver cell line, clone 9. Communication was observed and quantitated with microspectofluorometric and image analysis techniques following scrape loading of the cells with lucifer yellow. Lucifer yellow migrated as far as ten cells away from the scraped edge in control populations. Two minute TPA (25-50 micrograms/ml) treatment inhibited dye movement such that the dye remained mainly in the cells at the cut edge. The TPA-induced inhibition of cell-cell communication could be partially blocked by 15 min treatment of the cell populations with the PKC inhibitors trifluoperazine (30 micrograms/ml), staurosporine (2 x 10(-8) or 2 x 10(-6) M), sangivamycin (15 or 200 microM), or a PKC inhibitor peptide (20 micrograms/ml) scraped in at the same time as lucifer yellow. Normal communication was observed in cultures treated only with PKC inhibitors. Lower concentrations of TPA (50 ng/ml-1 micrograms/ml) used for 2 min did not inhibit dye communication. Our results demonstrate the phorbol ester induced interruption of cell-cell communication. The inhibition of PKC by inhibitors eliminates the effect of TPA on communication. Our data are consistent with a role of PKC in the control of junctional communication. PMID- 8417618 TI - Cellular proliferation in the anterior pituitary gland of normal adult rats: influences of sex, estrous cycle, and circadian change. AB - Proliferative activity of the anterior pituitary gland in 10 week-old male and female rats under normal conditions was investigated by counting mitotic figures and using single and double immunostaining of 5-bromo-2'-deoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA), and six pituitary hormones. To determine which proliferative changes depend on the estrous cycle and circadian changes, respectively, six groups of female and two groups of male rats were studied at various times of day. Additionally, BrdU-incorporated cells were further classified by the six types of hormones they contained, or as immunonegative cells. Cell proliferative activity in the females fluctuated drastically with the highest activity in estrus and the lowest in diestrus. In the males, proliferative activity was at a relatively low level, and was similar to that in females in proestrus or early estrus, with the greater activity at night. Identified by their pituitary hormones, the distribution of the proliferating cells was almost the same in each sex, with prolactin (PRL) cells accounting for the highest proportion, followed by growth hormone (GH) cells, and adrenocorticotropic hormone (ACTH), luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH) cells. These percentages agreed well with previously reported levels of cell types among all pituitary cells of the rat. It is therefore suggested that the life span and cycle of rat pituitary cells does not differ among cell types. In another test, male and female rats were given BrdU continuously via an osmotic pump for 8 days to compare cell proliferative activity between sexes, exclusive of the influence of estrous cycle and circadian changes. In this way, we were able to demonstrate that the cumulative incorporation of BrdU in females was consistently twice as high as in males over a constant period of time, and to conclude that cell renewal occurs at a doubled rate in the pituitary of female rat. PMID- 8417619 TI - Fate of odontoblasts and blood capillaries in the incisal region of the rat incisor pulp. AB - Transmission electron microscopy of thin sections of the rat incisor pulp revealed that in the middle region of the incisor there were fenestrated capillaries in the "predentinal plexus" and that this region contained the tallest odontoblasts. The odontoblasts gradually became shortened in the incisal part of this region; the fenestrated capillaries in the predentinal plexus changed to continuous type capillaries. Almost all the odontoblasts had degenerated near the incisal end of the tooth. The predentinal plexus disappeared in this region, but the "subodontoblastic capillary plexus" persisted. In a specific region just beneath the worn incisal end, numerous macrophages and polymorphonuclear neutrophils appeared and scavenged the degenerating cells, possibly including the odontoblasts. PMID- 8417620 TI - Quantitative morphological analysis of the pituitary gland in protein-calorie malnourished rats. AB - A quantitative analysis of the pituitary gland was conducted to ascertain the effects of protein-calorie malnutrition on the morphology of the somatotrophs, gonadotrophs, thyrotrophs, and corticotrophs. Male rats were fed a low, 8% protein diet from 20 to 50 days of age, while their age-matched controls were given a diet containing 27% protein. The hypophyses were then processed for light microscopic immunocytochemical staining using antibodies to growth hormone, the beta subunits of luteinizing hormone and thyroid stimulating hormone, and adrenal corticotrophic hormone. The number of each cell type along with an evaluation of the cell, cytoplasmic, and nuclear areas was conducted using a computerized image analyzer. All of these parameters were reduced significantly in the somatotrophs as a result of the low protein diet, while in the gonadotrophs, the cell, cytoplasmic, and nuclear areas were similarly affected. Smaller cell number, cell area, and nuclear area were noted in the corticotrophs of the malnourished animals, while in the thyrotrophs, only the cell and nuclear areas were reduced. The data demonstrate that each pituitary cell type responds in a unique manner to undernutrition. PMID- 8417621 TI - Morphometric analysis of the pelvis in mice treated neonatally with tamoxifen. AB - The pelves of male and female C57BL/Tw mice given five daily injection of 100 micrograms tamoxifen, 50 micrograms dihydrotestosterone (DHT), or 2 micrograms diethylstilbestrol (DES) from the day of birth were examined morphometrically and histomorphometrically. Total areas of the pelvis, ilium, ischium, and pubis were significantly smaller in neonatally tamoxifen-treated mice than in the controls. There was no significant difference in length of the ischium between tamoxifen treated and control mice of both sexes. However, lengths of ilium and pubis, and widths of ilium, pubis, and ischium in tamoxifen-treated male and female mice were significantly smaller than in the respective controls. In contrast, neonatal treatment with DHT or DES did not affect the shape of the pelvis of either sex. In the neonatally tamoxifen-treated females, the number of osteoblasts and osteoclasts per 200 microns trabecular surface length and per 10,000 microns2 subperiosteal area of pubic bone section was smaller than in the controls. Inhibition of ossification persisted in the junction of the pubis and ischium of pelves treated with tamoxifen in vitro. These results suggest that neonatally administered tamoxifen mainly retards the growth of the ilium and pubis in mice by changing the activities of osteoclasts and osteoblasts, and that tamoxifen acts directly on the neonatal mouse pubis to inhibit its ossification. PMID- 8417622 TI - Development of the liver in the chicken embryo. II. Erythropoietic and granulopoietic cells. AB - Hepatic hemopoiesis is apparent in the chicken embryo on day 7 of incubation (Hamburger and Hamilton Stage 30), and a peak in hemopoietic activity occurs on day 14 (Stage 40). During this period, the differentiation of hemopoietic cells was examined by light microscopy and by transmission and scanning electron microscopy. Glycol methacrylate sections were used in lieu of smears to study hemopoietic cells, thus minimizing the problems of cell shrinkage and rupture. The sections were superior to smears for close examination of nuclear and cytoplasmic morphologies and for precise localization of hemopoietic cells to intravascular and extravascular sites. The avian liver is involved directly with erythropoiesis and granulopoiesis only. Erythropoietic cells, occurring in intravascular and extravascular locations, appear throughout the time frame examined. Blood islands with granulopoietic cells were not observed until days 8 9 (Stage 35). Granulopoiesis in the liver produces only eosinophilic leukocytes. Individual granulopoietic cells appear first in the connective tissue sheaths of hepatic vessels, and these cells subsequently congregate into blood islands. Endothelial cells of the sinusoidal linings, through asymmetric divisions, frequently release daughter cells into the circulation, and Kupffer cells are actively engaged in phagocytosis of erythrocytes. From a comparative standpoint, the elements deemed critical to hemopoiesis in the mammalian liver--prehepatocyte population, hepatic vasculature, and compartments for stem cell differentiation- may not hold the same importance in the bird, owing to an inordinate reliance on intravascular hemopoiesis in this vertebrate class. PMID- 8417623 TI - Sites of renin production in fetal, neonatal, and postnatal Syrian hamster kidneys. AB - Renin is first observed in the 14-day fetal kidney. There is a sharp increase in the number of renin positive cells in the 15-day fetal kidney. Renin is located in the smooth muscle cells of arterioles, interlobular arteries, and branches of the renal artery. In the neonatal kidney, the amount of renin appears to be equal to that observed in the 15-day fetal kidney and is still located in the same blood vessels. In the 24-hour postnatal kidney, there is a sharp decrease in the total amount of renin. Renin positive cells are now observed at the vascular pole. In the 48-hour postnatal kidney, there is a sharp increase in the total amount of renin. Most of the renin positive cells are located at the vascular poles; however, a few renin positive cells are seen in the interlobular arteries. PMID- 8417624 TI - Scanning electron microscopy of developing primary endoderm during first 6 hours of incubation in the chick. AB - Development of primary endoderm in the domestic fowl (Gallus domesticus) is described in scanning electron microscopy (SEM) supplemented by transmission electron microscopy (TEM). Although complicated by great variability, the ventral surface of the blastoderm reveals this process during the first 6 hours of incubation. Primary endoderm arises (1) from the hypoblast, (2) from the margin of the area pellucida, and (3) from intervening portions of the area pellucida. The early hypoblast becomes several cells thick while individual cells are still spherical. TEM reveals a variety of immature cell junctions. During subsequent flattening of these cells into primary endodermal epithelium, numerous filopodia arise from their surfaces. These are 0.20-0.25 microns in diameter. They become long and branched, attaching to each other and to other cell bodies. Similar filopodial processes are present less conspicuously among cells in the margin of the area pellucida. Here, there is pseudopodial evidence that cells or cell sheets creep along the ventral surface of the epiblast. The filopodia disappear as cell flattening proceeds. The ventral surface of the exposed epiblast delaminates cells that become free after their exploratory filopodia and lamellipodia are put forth. Lateral contacts among cell bodies from the above three sources increase until a continuous epithelium is formed. The primary endoderm of the embryo, a simple squamous epithelium that separates the connective tissue space above from the gastrocoele below, is generated by these developmental events. PMID- 8417625 TI - Distribution, morphology, and central projections of mesencephalic trigeminal neurons in the frog Rana ridibunda. AB - The distribution, morphology, and central projections of the mesencephalic trigeminal neurons in the frog Rana ridibunda were studied with tracing techniques. Retrograde tracing with horseradish peroxidase (HRP) or the fluorescent tracer Fluorogold, and anterograde tracing by means of Phaseolus vulgaris leucoagglutinin, the fluorescent dye DiI, and HRP were used. The mesencephalic trigeminal nucleus (MesV) of Rana ridibunda is formed by a population of 100 to 125 unipolar or multipolar cells that are scattered on both sides of the rostral mesencephalic tectum. Subpopulations of Mes V cells were labeled after tracer application to ophthalmic, maxillary, and mandibular trigeminal branches, separately. Differences in the morphology and distribution of cells in these experiments were not evident but the number of neurons labeled via the maxillary nerve was always the highest. Mes V cells have a single central branch that courses caudally in the brainstem. At different levels, it bifurcates into a peripheral branch, which leaves the brain via the trigeminal root, and a descending branch, which terminates in a region in, or close to, the trigeminal motor nucleus and in a supratrigeminal location. The lack of a distinct somatotopy in the distribution of Mes V cells and the lack of projections caudal to the trigeminal motor nucleus as revealed in this study with a wide variety of tracers are in striking contrast to previous data provided for other amphibians. PMID- 8417626 TI - Heterogenous distribution of glycoconjugates in the kidney of dogfish Scyliorhinus caniculus (L.) with reference to changes in the glycosylation pattern during ontogenetic development of the nephron. AB - Eight fluorochrome-coupled lectins with different sugar specificities were applied to cryosections of dogfish kidney. Despite profound differences in renal architecture between elasmobranch fish and other vertebrates, the sequence of nephron segments as revealed by the lectin-binding pattern was rather similar to that of tetrapodes. Wheat germ agglutinin (WGA) bound to cell membranes of epithelial cells of glomeruli, proximal and distal tubules, their basement membranes, the collecting tubule, and epithelial cells. Among other broadly binding lectins were Ricinus communis agglutinin I (RCA-I), soybean agglutinin (SBA), peanut agglutinin (PNA), Lycopersicon esculentum agglutinin (LEA), and Jacalin, all of which marked proximal as well as distal portions of the renal tubule. Dolichos biflorus agglutinin (DBA) did not react with any renal structure. Ulex europaeus agglutinin I (UEA-I), which indicates the presence of alpha-L-fucose, very strongly and specifically marked single epithelial cells of the early distal nephron, all epithelial cells of the late distal tubule, the beginning of the collecting tubule in the mesial tissue zone, and single cells in the end portion of the collecting tubule in the lateral bundles. Binding of UEA-I to receptors of distal nephron cells could be useful for the identification of these cells in functional studies employing teased tubule and/or isolated cell preparations. Binding of UEA-I to dogfish kidney structures resembles staining with UEA-I conjugates of late distal tubules and collecting tubules in the kidneys of frog and other, higher vertebrates. Epithelial cells of early developmental stages showed, very rarely, binding sites for most lectin fluorochrome conjugates. A large number of lectin binding sites was observed in the extracellular matrix of fibroblast layers surrounding the early anlage and the S-shaped body. Lectin binding sites of the nephron epithelia appeared in a sequential manner in the next stages of development of the nephron. Ontogenetic and phylogenetic aspects of the merging region between nephron proper (late distal tubule) and collecting system (collecting tubule) are discussed. PMID- 8417627 TI - Ectoplasmic ("junctional") specializations in Sertoli cells of the rooster and turtle: evolutionary implications. AB - Ectoplasmic specializations are complex actin-containing structures found at certain sites of intercellular attachment in Sertoli cells. Current evidence indicates that these structures are a form of actin-associated adhesion junction. In the turtle (Pseudemys scripta) and rooster (Gallus domesticus) ectoplasmic specializations are known to occur adjacent to sites of attachment to elongate spermatids and are characterized by a layer of "loosely" cross-linked actin filaments that lies next to the plasma membrane. In the turtle, a cistern of endoplasmic reticulum is associated with the cytoplasmic face of the filament layer. We have found that, in fixed frozen sections of turtle and rooster testes, immunological probes for myosin II react with epithelial regions that also stain with probes for filamentous actin and that are known to be sites at which ectoplasmic specializations occur. Furthermore, when exposed to standard contraction buffers, the diameters of glycerinated ectoplasmic specializations of the turtle are statistically smaller than those of the same structures exposed to control buffers. We interpret these smaller diameters as being produced by the contraction of actin bundles within the ectoplasmic specializations. Our results indicate that ectoplasmic specializations in the rooster and turtle, unlike those in mammals, possess contractile properties. We speculate that ectoplasmic specializations in eutherian mammals may have evolved from actin-associated adhesion junctions in which the actin bundles were initially contractile and from which myosin II was secondarily lost. PMID- 8417628 TI - Epithelial reorganization and irregular growth following carbendazim-induced injury of the efferent ductules of the rat testis. AB - Occlusions of the rat efferent ductules were induced by an oral gavage of carbendazim and the capacity for regeneration or recanalization of the ductules was histologically evaluated between 2 and 70 days post-treatment. At 2 days post treatment, the efferent ductules were occluded by sloughed materials and showed various degrees of inflammation. Severely damaged epithelium showed few regenerative features at later intervals. On the other hand, epithelia with medium inflammation often exhibited irregular epithelial growth along the luminal contents or the formation of multiple abnormal ductules. These abnormal ductules were formed by migrated and original epithelia at the periphery of the occluded lumen at 16 days post-treatment, indicating attempted recanalization. At later time periods, 32 and 70 days post-treatment, the occluded original lumen was filled in by fibrotic connective tissue and surrounded by a series of abnormal ductules. These abnormal ductules were characterized by cuboidal epithelia, a small luminal diameter, fewer cilitated cells than normal, less developed organelles in the epithelial cells, and basal laminae of irregular thickness. However, there was no evidence that occluded ductules formed patent re connections via abnormal ductules. The results suggest that occluded efferent ductules have the ability to initiate epithelial regrowth and to form new ductules, but the newly formed ductules are abnormal and are not adequate to recover from azoospermia at least at 70 days post-treatment. PMID- 8417629 TI - Ultrastructure of epididymal interstitial reactions following vasectomy and vasovasostomy. AB - The response of the male reproductive tract to vasectomy includes inflammation of the interstitial tissue of the epididymis. The pathogenesis of epididymal interstitial reactions and characteristics of the responding cells were studied by electron microscopy in Lewis rats at intervals following bilateral vasectomy, vasectomy followed 1 month later by vasovasostomy, or sham operations. In areas of interstitial reaction, numerous macrophages, monocytes, lymphocytes, neutrophils, and plasma cells occupied the connective tissue. Macrophages, containing many lysosomes and vesicles, aggregated and assumed the appearance of epithelioid cells. Processes of adjacent macrophages interdigitated with one another and closely approached the surfaces of lymphocytes. Many plasma cells with distended rough endoplasmic reticulum appeared in the interstitium. The majority of animals in the vasectomy and vasovasostomy groups exhibited epididymal interstitial changes by 2-3 months; the cauda epididymidis was the region most often affected. The ultrastructural features were indicative of chronic granulomatous inflammation and were consistent with an immune response that includes antigen presentation by macrophages to lymphocytes, lymphocyte differentiation, and local antibody production by plasma cells. The nearly complete absence of sperm or recognizable parts thereof in the interstitial tissue in the areas of the reactions suggests that these lesions formed in response to soluble antigens leaking from the duct. Vasovasostomy was not effective in reversing or retarding epididymal inflammation at the intervals studied. PMID- 8417630 TI - Histological organization of the right and left atrioventricular valves of the chicken heart and their relationship to the atrioventricular Purkinje ring and the middle bundle branch. AB - In the avian heart the right and left atrioventricular (AV) valves not only exhibit their own special anatomical characteristics, but they also are in close proximity to the conduction system. The right AV valve is a single, spiral plane of myocardium, in remarkable contrast to the fibrous structure characteristic of the mammalian tricuspid valve. A ring of Purkinje tissue encircles the avian right AV orifice and connects to the muscular valve. The chicken has no crista supraventricularis, its right AV valve serving that function as well as opening and closing the right AV orifice. The left AV valve consists of three leaflets instead of the two typical of mammalian hearts. Its anterior and posterior leaflets are small; its large aortic (medial) leaflet merges with the bases of both the left and noncoronary cusps of the aortic valve by fibrous tissue, resembling that of the mammalian heart. However, unlike in mammals, there is a slim cylinder of continuous myocardium coursing parallel to this fibrous junction. This unusual arc of myocardium in the chicken serves to complete an entire subaortic ring of myocardium and is thus potentially capable of constricting the outflow tract of the chicken's left ventricle. The middle bundle branch connects with both the muscle arch and the AV Purkinje ring. Thus the myocardium in or near both AV valves (and the left ventricular outflow tract) in the chicken heart is so arranged that it may receive direct early activation from the conduction system. PMID- 8417631 TI - Comparative ultrastructural morphometric analysis of atrial specific granules in the bat, mouse, and rat. AB - Size, incidence, and volume density of atrial specific granules (ASG) in right atrial cells from five animals each of the rat (average weight 210 g), mouse (average weight 28 g), fruit-eating bat Megaloglossus woermanni (BMW; average weight 35 g), and the insect-eating bat Pipistrellus pipistrellus (BPP; average weight 6 g) have been compared via ultrastructural morphometry. In all three parameters of granule measurement, significantly higher figures were obtained in the rodents than in the bats. However, between the rat and the mouse, as also between the two species of bats, no significant differences were noted in any of the measurements. These results therefore do not support the prevalent view that the number and size of the granules decrease with increase in size of the animal species. The low content of ASG in atrial cells of the bats is probably an indication of low demand for the natriuretic hormone of the granule, because, in such animals, and particularly in flight, conservation of fluid and electrolytes is of paramount importance. This suggests that granule content is adapted to fluid and electrolyte regulation in relation to the functional capacity of the animal. We also observed ASG-like structures in endothelial cells of capillaries of bat tissue but not in rodents. The function of these granules or whether or not they represent atrial specific ones is not clear from the present study. PMID- 8417632 TI - Rabbit ductus arteriosus during development: anatomical structure and smooth muscle cell composition. AB - The anatomical structure as well as the smooth muscle cell (SMC1) composition of the ductus arteriosus (DA) were studied in rabbits ranging in age from 29 days of gestation to 20 days after birth. Computer-assisted, three-dimensional reconstructions of hematoxylin-eosin stained serial cryosections from ductus arteriosus-aorta (DA-AO) junctures revealed that DA in animals near term is separated from the aorta by a "septumlike" structure that is continuous with the aortic wall. Two days after birth, obliteration of DA is almost complete, and a small "pocketlike" cavity appears in the pre-existing site in which DA merged into the aorta. This small cavity in the aortic arch was still evident in the large majority of animals examined even 20 days after birth, as also demonstrated by scanning electron microscopy. At this time period DA consisted of a central, fibrotic region surrounded by several layers of SMC (the ligamentum arteriosum, LA) and ended within the aortic media just above the small cavity, forming a round "scar." Vascular SMC composition of DA during closure was examined by means of indirect and double immunofluorescence procedures, using a panel of monoclonal antibodies against some cytoskeletal and cytocontractile proteins (vimentin, desmin, smooth muscle (SM), and nonmuscle (NM) myosin isoforms). "Intimal cushions" were particularly evident from 5 hr after birth and were found to be desmin-negative, homogenously reactive for vimentin and NM myosin, and heterogeneously stained with anti-SM myosin antibody. In SMC subjacent to the "intimal cushions," distribution of vimentin and SM myosin was homogeneous, whereas the one of desmin and NM myosin content was heterogeneous. The cytoskeletal and cytocontractile protein content displayed by SMC during the closure of DA is similar to that of "intimal thickening" found in some pathological conditions of the arterial wall in adult rabbits. Completation of DA closure (day 2) was accompanied by the disappearance of cellular heterogeneity in myosin isoform distribution in both the "intimal cushions" and the underlying media. These results give new insights into: (1) the structure of DA-AO juncture, which can be relevant to the physiology of blood circulation in the fetus, and (2) the phenotypic similarity of vascular SMC populations involved in the formation of "intimal cushions" and "intimal thickening." PMID- 8417633 TI - Anaphylactic shock after insect-sting challenge in 138 persons with a previous insect-sting reaction. AB - OBJECTIVE: To study the rate and severity of anaphylactic reaction in relation to plasma levels of cardiovascular mediators in persons with a history of insect sting anaphylactic shock who were rechallenged with a sting by the same insect. DESIGN: A cohort study with measurements before and after intentional sting challenge. SETTING: Intensive care unit of an 830-bed general hospital, a national center of insect-sting anaphylaxis in The Netherlands. PATIENTS: A total of 138 patients referred after a previous anaphylactic reaction to a Hymenoptera sting; and 8 volunteers. MEASUREMENTS: Signs of anaphylaxis and plasma levels of catecholamines and angiotensins. MAIN RESULTS: Only 39 of 138 (28%) of patients with a previous insect-sting anaphylactic reaction developed anaphylactic symptoms after sting challenge. Values of cardiovascular mediators and mean arterial pressure did not differ after the challenge from initial values in the volunteers or in the patients with a mild or no reaction after challenge. In the 17 patients with anaphylactic shock, mean arterial pressure decreased from 97 +/- 11 (mean +/- SD) to 65 +/- 17 mm Hg (P < 0.001), epinephrine levels rose from a median of 0.3 nmol/L (range, 0.2 to 2.3 nmol/L) to 2.5 nmol/L (0.2 to 35.7 nmol/L; P < 0.05), norepinephrine from 1.5 nmol/L (0.5 to 6.7) to 5.9 nmol/L (1.6 to 30.9 nmol/L; P < 0.01), and angiotensin II from 61 pmol/L (7 to 217 pmol/L) to 105 pmol/L (11 to 286 pmol/L; P < 0.01), all within 5 minutes after the onset of anaphylactic symptoms. The rise of these mediators correlated with the drop in blood pressure (P < 0.001). Dopamine and angiotensin I levels did not change in any participants. CONCLUSIONS: A recurrent insect-sting anaphylactic reaction occurred in only 28% of patients with a previous reaction. During this recurrent reaction, plasma levels of endogenous epinephrine, norepinephrine, and angiotensin II rose in relation to hypotension. PMID- 8417634 TI - What is the benefit of increasing the sulfonylurea dose? AB - OBJECTIVE: To study the clinical benefit of increasing the dose of the sulfonylurea, glipizide, from 10 to 40 mg per day. DESIGN: A placebo-controlled, double-blind, cross-over study. SETTING: An outpatient clinic at the Helsinki University Hospital, Finland. PATIENTS: Twenty-three patients with noninsulin dependent diabetes mellitus. METHODS: Patients were given glipizide in three different dose schedules for 3 months each: 10 mg in the morning or 10 mg or 20 mg in the morning and the evening. Glycemic control was followed by HbA1c measurements and blood glucose monitoring at home. Beta cell function was assessed by measuring insulin responses to a test meal. RESULTS: Mean home monitored blood glucose was 12.4 mmol/L during placebo treatment, and it was 9.6, 9.2, and 8.9 mmol/L after treatment with glipizide, 10, 20, or 40 mg, respectively. The levels of blood glucose and HbA1c differed after all treatment groups from placebo (P < 0.001) but not among themselves. The insulin response to a test meal was greatest after 10 mg of glipizide and weakest after 40 mg/d (P = 0.02 compared with the 10-mg dose). All treatments stimulated insulin secretion more than placebo (P < 0.001). CONCLUSIONS: Increasing the glipizide dose to more than 10 mg once daily produces little or no benefit and may reduce beta-cell function. PMID- 8417635 TI - Increased mortality with gallstone disease: results of a 20-year population-based survey in Pima Indians. AB - OBJECTIVE: To determine if gallstone disease is associated with an increased risk for malignancy and higher total mortality in Pima Indians. DESIGN: Inception cohort. SETTING: American Indian community. PARTICIPANTS: Age- and sex-stratified random population-based sample. MEASUREMENTS: Between 1966 and 1969, an age- and sex-stratified random sample of Pima Indians from the Gila River Indian Community in Arizona was examined to identify evidence of gallstone disease defined as either gallstones (oral cholecystography) or previous cholecystectomy. During 20 years of follow-up, deaths were recorded and underlying causes of death, according to death certificates, were determined. RESULTS: Among 383 persons with known gallbladder status, 186 (49%) died: 133 among the 222 persons with gallstone disease and 53 among the 161 without. The overall death rate was higher in persons with gallstone disease than in those with normal gallbladders. The age and sex-adjusted death rate ratio was 1.9 (95% Cl, 1.3 to 2.7). Furthermore, the death rate attributed to malignancies was 6.6 times (Cl, 1.3 to 33.1) as high in persons with gallstone disease as in those with normal gallbladders. Of the 20 fatal malignancies in persons with gallstone disease, 11 occurred in the digestive tract, of which six involved the gallbladder or bile ducts. CONCLUSIONS: Increased cancer mortality and total mortality were found in Pima Indians with gallstone disease. Although plausible explanations exist for the increased cancer mortality, the increased death rates due to other causes are unexplained. Whether cholecystectomy would change this risk is unknown. PMID- 8417636 TI - The natural history of asymptomatic hepatitis B surface antigen carriers. AB - OBJECTIVE: To assess the long-term outcome in hepatitis B surface antigen (HBsAg) carriers who have normal liver function tests, focusing on survival and the development of severe liver disease and hepatocellular carcinoma. DESIGN: Cohort study with a mean follow-up of 130 months. SETTING: Liver clinic of a referral center. PATIENTS: Ninety-two HBsAg-positive blood donors with normal liver function tests. MEASUREMENTS: Histologic evaluation of liver specimens at baseline; clinical, biochemical, and serologic follow-up; and repeat liver biopsy if clinically indicated or after 10 years of follow-up. RESULTS: At baseline, 69 subjects had normal histologic findings or only minor abnormalities, 18 had chronic persistent hepatitis, and 5 had mild chronic active hepatitis. Serum enzyme levels remained normal in 58 of 68 patients who had regular follow-up. Three patients had biochemical changes consistent with hepatitis B virus (HBV) infection; in one of these patients, a later histologic evaluation showed progression to chronic active hepatitis. One patient developed alcoholic cirrhosis. Six other patients had mild or transient transaminase elevations, with no evidence of HBV replication, hepatitis D virus infection, hepatitis C virus (HCV) infection, or histologic deterioration. Liver histologic findings also remained unchanged in 21 patients who showed no biochemical changes during 10 years of follow-up and consented to have repeated liver biopsy. Ten patients showed loss of HBsAg; 2 of these patients acquired antibody to hepatitis B surface antigen (anti-HBs). All patients who did not have regular follow-up, except 1, were interviewed by telephone during 1990: All denied having liver disease. No patients developed hepatocellular carcinoma. CONCLUSIONS: Italian HBsAg carriers with initially normal liver function tests have an excellent prognosis: Delta superinfection is infrequent and the risk for developing hepatocellular carcinoma is low. PMID- 8417637 TI - Possible bromocriptine-induced myocardial infarction. PMID- 8417638 TI - The risk of determining risk with multivariable models. AB - PURPOSE: To review the principles of multivariable analysis and to examine the application of multivariable statistical methods in general medical literature. DATA SOURCES: A computer-assisted search of articles in The Lancet and The New England Journal of Medicine identified 451 publications containing multivariable methods from 1985 through 1989. A random sample of 60 articles that used the two most common methods--logistic regression or proportional hazards analysis--was selected for more intensive review. DATA EXTRACTION: During review of the 60 randomly selected articles, the focus was on generally accepted methodologic guidelines that can prevent problems affecting the accuracy and interpretation of multivariable analytic results. RESULTS: From 1985 to 1989, the relative frequency of multivariable statistical methods increased annually from about 10% to 18% among all articles in the two journals. In 44 (73%) of 60 articles using logistic or proportional hazards regression, risk estimates were quantified for individual variables ("risk factors"). Violations and omissions of methodologic guidelines in these 44 articles included overfitting of data; no test of conformity of variables to a linear gradient; no mention of pertinent checks for proportional hazards; no report of testing for interactions between independent variables; and unspecified coding or selection of independent variables. These problems would make the reported results potentially inaccurate, misleading, or difficult to interpret. CONCLUSIONS: The findings suggest a need for improvement in the reporting and perhaps conducting of multivariable analyses in medical research. PMID- 8417639 TI - Hazards of hospitalization of the elderly. AB - For many older persons, hospitalization results in functional decline despite cure or repair of the condition for which they were admitted. Hospitalization can result in complications unrelated to the problem that caused admission or to its specific treatment for reasons that are explainable and avoidable. Usual aging is often associated with functional change, such as a decline in muscle strength and aerobic capacity; vasomotor instability; reduced bone density; diminished pulmonary ventilation; altered sensory continence, appetite, and thirst; and a tendency toward urinary incontinence. Hospitalization and bed rest superimpose factors such as enforced immobilization, reduction of plasma volume, accelerated bone loss, increased closing volume, and sensory deprivation. Any of these factors may thrust vulnerable older persons into a state of irreversible functional decline. The factors that contribute to a cascade to dependency are identifiable and can be avoided by modification of the usual acute hospital environment by deemphasizing bed rest, removing the hazard of the high hospital bed with rails, and actively facilitating ambulation and socialization. The relationships among physicians, nurses, and other health professionals must reflect the importance of interdisciplinary care and the implementation of shared objectives. PMID- 8417640 TI - Informed consent. PMID- 8417641 TI - Now is the time: physician involvement in health care reform. PMID- 8417642 TI - Perverse incentives, statesmanship, and the ghosts of reforms past. PMID- 8417643 TI - Leukocyte esterase tests detect pyuria, not bacteriuria. PMID- 8417644 TI - Leukocyte esterase tests detect pyuria, not bacteriuria. PMID- 8417645 TI - Risk factors for amputation in diabetics. PMID- 8417646 TI - Publication criteria for statistical prediction models. PMID- 8417647 TI - "Fiancees'" Graves disease. PMID- 8417649 TI - CME credits for cleaning the "med closet". PMID- 8417648 TI - Calculating body mass index. PMID- 8417650 TI - Access to health care. PMID- 8417651 TI - Access to health care. PMID- 8417652 TI - Access to health care. PMID- 8417653 TI - Access to health care. PMID- 8417654 TI - Access to health care. PMID- 8417655 TI - Access to health care. PMID- 8417656 TI - Access to health care. PMID- 8417657 TI - Bovine internal mammary artery as a conduit for coronary revascularization: long term results. AB - Graft patency after coronary artery bypass grafting depends largely on the choice of conduit. Because an increasing number of patients have insufficient or poor quality autologous material, there is a need for a suitable synthetic graft that is readily available and easy to handle and that has good long-term patency. Early results suggest that the bovine internal mammary artery graft may meet these criteria. We have used a total of 26 such grafts in 18 patients. Postoperative angiography has been performed in 19 grafts in 14 patients, 3 to 23 months after operation; of these grafts, 3 are currently patent (15.8%, compared with 85.7% and 75.0% patency for native internal mammary artery and saphenous vein grafts in the same patients). We report the results of clotting studies and an analysis of lipid status. These patients do not, however, appear to represent any atypical group, either in terms of coagulopathy, native coronary artery size, or the type of vessel disease. Nevertheless, our poor results contrast markedly with the early enthusiasm reported from other centers. PMID- 8417658 TI - Perfusion pressure control by adenosine triphosphate given during cardiopulmonary bypass. AB - Administration of exogenous adenosine triphosphate (ATP) as a vasodilator during cardiopulmonary bypass was assessed in consecutive adult patients (n = 24) who demonstrated a high arterial perfusion pressure (mean, > 90 mm Hg). The action of ATP was characterized by rapid induction and stabilization of the blood pressure level. The dose of ATP ranged from 0.68 to 2.68 mg/min. Within 1 minute after the administration, there was a significant reduction in the perfusion pressure from 102 +/- 18 mm Hg (mean +/- standard deviation) to 72 +/- 19 mm Hg. The ATP was then able to maintain the desired pressure of 69 +/- 12 mm Hg at 5 minutes, 67 +/ 12 mm Hg at 10 minutes, and consistent values thereafter. After the ATP administration was discontinued, there was a prompt recovery of pressure without bradyarrhythmia. The frequency and amount of inotropes used were consistent with the control group (n = 26). Although the administration of ATP reduced the increase in serum catecholamine concentration, there were no significant changes in other vasoactive mediators (eicosanoid, angiotensin II, endothelin) between the two groups during cardiopulmonary bypass. There was neither an accumulation of metabolic products (uric acid, phosphate) nor a decrease in the level of divalent cation (Ca2+), which is observed when the cations combine with phosphates or adenosine nucleotides. This study confirmed the efficacy and safety of ATP infusion during cardiopulmonary bypass. PMID- 8417659 TI - Effects of cryothermia during cold cardioplegia on epicardial and intramural coronary arteries. AB - Cryosurgery is playing an increasingly important role in the surgical treatment of various supraventricular and ventricular tachyarrhythmias. The short-term and long-term effects of cryogenic injury on epicardial and intramural coronary arteries were studied in 22 dogs. Transmural cryolesions encompassing a posterolateral branch of the circumflex coronary artery were produced in the inferior left ventricular wall during extracorporeal circulation and cold cardioplegic arrest. The mean epicardial cryolesion area +/- standard deviation was 10.4 +/- 1.8 cm2. The mean epicardial coronary artery diameter +/- standard deviation measured 1.2 +/- 0.4 mm. At 6 hours, no important structural changes were noted in any of 6 dogs. At 48 hours, 1 of 2 epicardial coronary arteries showed recent thrombus. At 1, 3, and 6 months, the epicardial coronary arteries were occluded due to thrombosis and intimal hyperplasia in 13 of 14 dogs. A limited degree of recanalization was observed. At all follow-up intervals, the intramural coronary arteries exhibited a histologic pattern similar to that of the epicardial coronary arteries. It is concluded that the exposure of major epicardial coronary arteries to cryoinjury during cold cardioplegic arrest should be avoided where possible. PMID- 8417660 TI - Improved tracheal allograft viability in immunosuppressed rats. AB - Airway ischemia has been a common cause of morbidity and mortality in clinical lung transplantation. The present study examined the effects of cyclosporin A (CsA) and methylprednisolone (MP) on the viability of the devascularized trachea after heterotopic transplantation and omentopexy. Thirty-six tracheal segments were harvested from 18 donor Lewis rats, wrapped in omentum, and heterotopically implanted into the abdomen of recipient rats. Tracheal segments were randomly allocated into one of six recipient groups (n = 6): Lewis syngeneic controls, and five groups of Brown Norway recipients, receiving either no treatment, CsA alone (5 mg.kg-1.day-1 or 15 mg.kg-1.day-1), or CsA in combination with MP (5 mg CsA + 1 mg MP per kg/day or 15 mg CsA + 2 mg MP per kg/day, respectively). After 14 days, the tracheal segments were histologically evaluated. Epithelial thickness and the degree of epithelial regeneration were significantly different (p < 0.05) between the syngeneic control group and the untreated Brown Norway group. Without immunosuppression there was virtually no epithelium, whereas low-dose immunosuppression yielded intermediate viability, and with high dose CsA and MP we observed improved tracheal viability. In this high-dose group, the epithelium was thicker than in even the syngeneic control group. These results indicate that, in heterotopic tracheal allografts, viability may be improved with an optimum combination of CsA and MP. PMID- 8417661 TI - Argon beam coagulation compared with cryoablation of ventricular subendocardium. AB - The Argon Beam Coagulator uses radiofrequency energy to excite argon gas that may be used for ventricular ablation. The effects of power level and number of applications of the Argon Beam Coagulator were compared wtih cryothermia. Ten mongrel dogs underwent cardiac extirpation. The endocardial surfaces of 5 hearts were used for the creation of lesions using the Argon Beam Coagulator at five power levels with either one or two applications. Five hearts were used for endocardial and epicardial lesions using cryothermia (15-mm-diameter probe at -70 degrees C) for 1, 2, 3, or 4 minutes. The Argon Beam Coagulator lesions showed an increase in depth with increasing power levels (2.25 +/- 1.05 mm at 50 W to 6.64 +/- 0.75 mm at 150 W) and number of applications (maximum depth of 6.64 +/- 0.75 mm with one application, 11.2 +/- 1.1 mm with two applications). Cryothermia lesions were similar in depth regardless of duration or site of application (range, 6.1 to 10.2 mm). Both techniques resulted in homogeneous and well demarcated lesions. These data show that the Argon Beam Coagulator results in discrete endocardial lesions, which may be created quickly and reproducibly. This may be a useful alternative for the operative ablation of endocardial scar in the treatment of ventricular tachycardia. PMID- 8417662 TI - Noncardiogenic pulmonary edema complicating lung resection. AB - Postresectional pulmonary edema is a rare but potentially fatal complication of thoracic operations. In a retrospective study of 402 lung resections we have identified 11 cases of postresectional, noncardiogenic pulmonary edema. We have analyzed the individual data to test recognized hypotheses regarding this condition. Pulmonary edema occurred in 5.1% of right pneumonectomies, 4.0% of left pneumonectomies, and 1% of all lobectomies. In 2 patients the symptoms occurred immediately after operation; in the other 9 the mean interval to diagnosis was 43.4 hours. All 11 patients were in a positive fluid balance in the first 24 hours after operation (mean, 20.8 +/- 9.1 mL/kg). However, there was no significant difference between this value and the respective values for control groups of 20 patients having pneumonectomies and 20 patients having lobectomies in whom pulmonary edema did not develop. Our findings differ from other reported series in that perioperative fluid overload was not found to be a significant contributory factor in the development of postresectional pulmonary edema. We discuss other possible mechanisms for this phenomenon. PMID- 8417663 TI - Comparison of two experimental models for assessment of cardiac preservation. AB - Previous studies from this institution using human cell cultures have suggested that University of Wisconsin solution is preferred for prolonged hypothermic storage for cardiac transplantation. The primary objective of this study was to evaluate the effectiveness of extended cardiac preservation with University of Wisconsin solution by assessing the time-related changes of purine metabolites using two different models of cold storage. Isolated rat hearts (n = 6/group) or human ventricular myocyte cultures (n = 7 dishes/group) were assessed after 0, 6, 12, and 24 hours in University of Wisconsin solution at 0 degrees C using high performance liquid chromatography. Adenosine triphosphate content decreased from 18.1 +/- 5.4 to 9.6 +/- 2.7 mumol/g dried weight by 12 hours and to 1.0 +/- 0.6 mumol/g by 24 hours (p < 0.0001 by analysis of variance) in the rat model. Adenosine triphosphate content decreased from 0.64 +/- 0.42 to 0.14 +/- 0.11 nmol/micrograms DNA at 6 hours and to 0.04 +/- 0.03 nmol/micrograms DNA by 24 hours (p < 0.00001) in the cardiomyocytes. Inosine monophosphate content increased from 0.1 +/- 0.2 to 10.8 +/- 1.0 by 24 hours (p < 0.0001) in the rat studies. Inosine monophosphate values tended to increase up to 12 hours (p = 0.06) in the cell cultures and then declined. Adenosine concentration increased from 0.3 +/- 0.3 to 2.3 +/- 0.9 mumol/g at 6 hours and declined thereafter (p < 0.0005) in the rodent hearts. Adenosine concentration increased from 0.03 +/- 0.02 to 1.53 +/- 0.72 nmol/micrograms DNA at 6 hours (p < 0.0001) in the cardiomyocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417664 TI - Distal aortic arch aneurysmectomy and coronary revascularization through a left thoracotomy. AB - A successful single operation of a distal aortic arch aneurysm and coronary artery disease through a left lateral thoracotomy using a simple hypothermic retrograde cerebral perfusion technique for cerebral protection in a 64-year-old man is reported. During ventricular fibrillation accompanying cooling to 15 degrees C, a left internal thoracic artery was anastomosed with the left anterior descending coronary artery, and the aneurysm was replaced with a patch during hypothermic retrograde cerebral perfusion. PMID- 8417665 TI - Extracorporeal membrane oxygenation for severe heart failure after Fontan operation. AB - A 10-year-old boy with tricuspid atresia and Glenn anastomosis underwent a modified Fontan operation. After the operation, the pressure in the inferior vena cava increased, leading to oliguria and ascites. After the creation of continuity between the superior vena cava and the inferior vena cava to reduce the pressure gradient, there remained an elevated right atrial pressure. Six days of extracorporeal membrane oxygenation effectively stabilized his hemodynamics and organ function and allowed an excellent outcome. PMID- 8417666 TI - Thoracoscopic drainage and resection of giant mediastinal cyst. AB - Video thoracoscopy is being used with increasing frequency by thoracic surgeons. Although initially used primarily for simple diagnostic purposes, improved instrumentation and evolving expertise have allowed for the performance of increasingly complex therapeutic procedures. We present a case of a giant subcarinal bronchogenic cyst in an elderly man with multiple medical problems. The cyst nearly occluded the bronchus intermedius and resulted in recurrent respiratory tract infections. Using video thoracoscopy, the cyst was drained and the wall of the cyst was resected successfully with minimal concomitant morbidity. PMID- 8417667 TI - Combined thrombolytic therapy for prosthetic mitral valve thrombosis. AB - Prosthetic valve thrombosis is associated with high mortality. The treatment of choice remains operation. This is a case report of the successful combination therapy of tissue plasminogen activator and urokinase for an isolated thrombosed prosthetic mitral valve in a postpartum patient in whom operation was thought to carry an unacceptable risk. Combined thrombolytic therapy or therapy with a single agent with a long half-life and a prolonged infusion time is suggested as an emergent treatment option for prosthetic mitral valve thrombosis. PMID- 8417668 TI - Free splenic artery used in aortocoronary bypass. AB - Many alternative bypass conduits for coronary revascularization have been used since the introduction of the saphenous vein. The internal mammary artery has demonstrated superior long-term patency rates compared with vein grafts. Other arterial grafts previously investigated include the right gastroepiploic artery, inferior epigastric artery, radial artery, and splenic artery. This case reports bypass using a free splenic artery and a pedicled right gastroepiploic artery, each with successful postoperative patency. PMID- 8417669 TI - Treatment of incessant reciprocating atrioventricular tachycardia by a closed heart surgical technique. AB - The case of a 40-year-old woman having disabling symptoms of incessant atrioventricular reciprocating tachycardia is reported. She underwent surgical therapy by a closed heart technique. Operative technique and the advantages of the epicardial approach in the present case are detailed. PMID- 8417670 TI - Management of a mediastinal cyst causing hyperparathyroidism and tracheal obstruction. AB - A 78-year-old man with a history of parathyroidectomy presented with hyperparathyroidism and tracheal obstruction. After excision of the mediastinal parathyroid cyst, his symptoms were completely relieved. The diagnosis, investigations, and management are discussed. PMID- 8417671 TI - Multiple tumor emboli after pneumonectomy. AB - Systemic arterial tumor embolization is a rare complication after pulmonary resection. It may occur with primary or metastatic pulmonary malignancies, and is more frequently associated with pneumonectomy than with lesser pulmonary resections. Despite the rarity of this complication, tumor emboli should be considered in the etiology of extremity, cerebral, or multiorgan ischemia occurring during the perioperative period in patients undergoing pulmonary resections for malignancy. PMID- 8417672 TI - Preoperative management of neonatal tetralogy of Fallot with absent pulmonary valve syndrome. AB - In tetralogy of Fallot with absent pulmonary valve syndrome, aneurysmal enlargement of the central pulmonary arteries can be associated with severe bronchial stenosis and respiratory distress in the neonatal period. The ventilatory status of these patients must be stabilized before diagnostic and therapeutic interventions can be performed. This report describes the experience with prone positioning and emergency sternotomy in the preoperative care of a neonate. PMID- 8417673 TI - Retrograde cardioplegia cannulation during cardiopulmonary bypass. AB - A simplified method of transatrial cannulation of the coronary sinus during cardiopulmonary bypass, with the heart completely decompressed, is described. The technique is easy to adopt and inherently safe because the catheter is guided toward the great cardiac vein visually, rather than by manual palpation. PMID- 8417674 TI - Simplified method for delivering normothermic blood cardioplegia. AB - We describe a simple technique for optimizing oxygen delivery during normothermic continuous blood cardioplegia. It involves the use of a minimal volume of cardioplegic agents, the infusion rate of which is adjusted so as to keep the heart arrested. The resulting enhancement of oxygen supply is marshalled from the maintenance of hematocrit values in the range of 0.25. PMID- 8417675 TI - Use of expanded polytetrafluoroethylene pericardial substitute with ventricular assist devices. AB - Expanded polytetrafluoroethylene membrane was used to reconstruct the pericardium after ventricular assist device insertion in 7 consecutive patients. One patient remains on support with the membrane in place and 6 have had the membrane removed. The expanded polytetrafluoroethylene membrane protected the device cannulas during repeat sternotomy and expedited the dissection of the heart and great vessels at the time of device removal. Reconstruction of the pericardium with expanded polytetrafluoroethylene membrane is recommended at the time of ventricular assist device insertion. PMID- 8417676 TI - Naming the bronchopulmonary segments and the development of pulmonary surgery. AB - An international agreement on bronchial nomenclature and anatomy was not reached until well after operations for bronchopulmonary segmental disease were well developed. R. C. Brock, in 1950, was the reporter of the efforts of The Thoracic Society of Great Britain to bring some order to this confused state. This Society delayed its action until an ad hoc committee made up of members from other countries and specialties met at the International Congress of Otorhinolaryngology in 1949. The anatomy and nomenclature of the bronchopulmonary segments was agreed upon. The Thoracic Society then accepted the report of the ad hoc committee. The system was followed closely by the first Nomina Anatomica in 1955. This report did not open new surgical vistas but was the marker indicating that pulmonary surgery was now mature. PMID- 8417677 TI - Solitary squamous cell papilloma of the trachea. AB - We report a case of solitary squamous cell papilloma of the trachea and review 55 other previously reported cases. A 69-year-old man was admitted to our hospital with hemoptysis. Bronchoscopy demonstrated a papillomatous tumor diagnosed as a squamous cell papilloma. A circumferential resection of the trachea with end-to end anastomosis was carried out. The patient's postoperative course was uneventful. From the review, there seems to be no considerable difference in the prognoses between the endoscopically and surgically managed groups. The following indications for surgical management of solitary tracheobronchial papilloma are proposed: (1) wide-based tumor, (2) poorly visualized tumor by endoscopy, (3) suspicion of malignant tumor, and (4) patients not suitable for long-term follow up. PMID- 8417678 TI - Conduits for coronary artery bypass. PMID- 8417679 TI - Retrograde warm blood cardioplegia. PMID- 8417680 TI - White matter lesions and heart operations. PMID- 8417681 TI - Direct mechanical ventricular actuation. PMID- 8417682 TI - Intraaortic balloon position. PMID- 8417683 TI - Ionescu-Shiley standard-profile and low-profile valves. PMID- 8417684 TI - Subacute cardiac rupture: repair with a sutureless technique. AB - Thirteen patients with ages between 53 and 74 years had development of free wall left ventricular rupture after a myocardial infarction (mean interval, 3.8 days). All patients showed clinical signs of cardiac tamponade. Diagnosis was established by bedside multiple pressure monitoring and echocardiography, which showed pericardial effusion with compression of the right ventricle. Cardiac catheterization was not performed. A new surgical technique was employed for the repair. After the pericardium was opened and cardiac tamponade was relieved, the myocardial tear was identified. A Teflon patch was applied over the area and glued to the heart surface with a surgical glue (cyanoacrylate). Cardiopulmonary bypass was not used except in a patient with a posterior tear. The method was consistently effective in controlling bleeding from the myocardial tear. All patients survived the operation and were discharged from the hospital a mean of 15 days after the operation. Follow-up extending up to 5 years (mean, 26 months) shows a 100% survival, 11 asymptomatic patients, and 2 patients with mild exertional angina. The technique is a simple, effective, and safe method for repair of subacute cardiac rupture and obviates the need for suturing on an infarcted ventricle. PMID- 8417685 TI - Thoracoscopic excision of bronchogenic cysts. PMID- 8417686 TI - Circulatory support 1991. The Second International Conference on Circulatory Support Devices for Severe Heart Failure. Patient selection. PMID- 8417687 TI - Circulatory support 1991. The Second International Conference on Circulatory Support Devices for Severe Cardiac Failure. Anticoagulation. PMID- 8417688 TI - Circulatory support 1991. The Second International Conference on Circulatory Support Devices for Severe Heart Failure. Infections--prophylaxis and treatment. PMID- 8417689 TI - Circulatory support 1991. The Second International Conference on Circulatory Support Devices for Severe Cardiac Failure. Management of secondary organ dysfunction. PMID- 8417690 TI - Circulatory support 1991. The Second International Conference on Circulatory Support Devices for Severe Cardiac Failure. Long-term biventricular assist. PMID- 8417691 TI - Circulatory support in infants and children. AB - Although the last decade has brought dramatic improvement in patient selection and postoperative management of adults and children undergoing advanced mechanical circulatory support, technological advances have been largely limited to the adult population. Intraaortic balloon pumps are technically feasible, but their efficacy has been questioned and their use has been limited in children. Over the last decade, extracorporeal membrane oxygenation has become the most commonly used method of mechanical circulatory support in children who have severe cardiac failure after cardiac operations. Additionally, a small number of infants and children have been supported with extracorporeal membrane oxygenation as a bridge to cardiac transplantation or as temporary support during reversible myocardial failure due to lesions such as viral myocarditis. More recently, a small number of pediatric patients have been supported with centrifugal and pneumatic devices, particularly after cardiac operations. Surprisingly, many of these patients did well with left ventricular support only. The overall children's survival rates in the myocardial recovery group are better than those in adults. However, current pediatric devices do not provide support for greater than a few weeks, making bridging to transplantation less feasible than in adults. PMID- 8417692 TI - Circulatory support with shock due to acute myocardial infarction. AB - Cardiogenic shock after acute myocardial infarction develops according to the amount of lost myocardium, function of remote myocardium, and the phenomenon of infarct expansion. Patients treated with mechanical support alone, without additional measures, have a mortality rate of 80%, the same as patients treated medically. Emergency angioplasty and emergency coronary artery bypass grafting can reduce mortality in certain subsets of patients to 40%. Patients with more severe shock and secondary organ dysfunction may be treated with mechanical bridging to transplantation with survival rates varying between 45% and 76%. Percutaneous support systems may be used to resuscitate a patient or to temporize, allowing time to perform diagnostic studies to determine if the patient is suitable for revascularization or heart transplantation. Intravenous enoximone may improve cardiac function as well and thus allow better decision making for further therapy. PMID- 8417693 TI - The biopump and postoperative circulatory support. PMID- 8417694 TI - Human cryopreserved homografts: electron microscopic analysis of cellular injury. AB - Twenty-five human cryopreserved valves with harvest-related warm ischemic times (WITs) ranging from 0 to 20 hours were studied using transmission electron microscopy to characterize the effects of harvesting and preservation on leaflet matrix cells. The valves were divided into seven groups on the basis of WIT and processed using standard transmission electron microscopic methods. Each cell (528 micrographs) was graded for reversible and irreversible cellular injury and subjected to a Cochran-Mantel-Haenszel trend analysis. Our results demonstrated a progression in cellular injury with increasing WIT. During the first 12 hours of warm ischemia, reversible cellular injury predominated (0.0%, 30.0%, 51.2%, 31.3%, 35.1%, 45.1%, and 40.0% at WITs of 0, 1, 2, 8, 12, 16, and 20 hours, respectively). A positive correlation (p < 0.0001) between increasing WIT and reversible cellular injury through the first 12 hours was observed. Minimal morphologic evidence of irreversible injury was noted in valves harvested with less than 12 hours of warm ischemia; however, after 12 hours there was a marked increase (0.0%, 0.0%, 4.7%, 2.4%, 2.7%, 31.4%, and 40.0% at WITs of 0, 1, 2, 8, 12, 16, and 20 hours, respectively) in irreversible cellular injury (p < 0.001 between 12 and 20 hours WIT). These data demonstrate a progression in cellular injury with increasing WIT. There was virtually no morphologic injury in valves with harvest-related WITs less than 2 hours and minimal irreversible cellular injury observed in valves exposed to 12 hours or less of warm ischemia. If cellular viability is critical to homograft durability then harvest-related warm ischemia may need to be restricted to 12 hours. PMID- 8417695 TI - Clinical experience with ventricular assist systems in Japan. AB - In Japan, five types of ventricular assist system, including the government approved National Cardiovascular Center type and the Tokyo University type, were applied to 202 patients as of September 30, 1991. Adult-sized ventricular assist systems were used in 194 patients and a pediatric one in 8. The major primary diagnosis was ischemic heart disease (52.5%). Sixty-three patients in cardiogenic shock with acute myocardial infarction were treated using a ventricular assist system, and the results were fairly good. The predominant indication was failure to be weaned from cardiopulmonary bypass (72.8%), and there was no case of bridging to heart transplantation. Left ventricular assist was performed in most patients (85.1%). The duration of ventricular assist system application ranged from 1 hour to 70 days, and the mean duration was 6.8 +/- 9.6 days. Duration of circulatory support had no relation to the results, and the longest support in survivors was 25 days. The weaning rate was 50.0%, and the survival rate was 26.2%. Main causes of death were heart failure, including unrecoverable heart failure, and multiple organ failure. It should be kept in mind that the decision to use a ventricular assist system should be made quickly, before major organs, including the heart itself, are irreversibly damaged. PMID- 8417696 TI - External pulsatile circulatory support. PMID- 8417697 TI - Internal pulsatile circulatory support. PMID- 8417698 TI - Nonpulsatile circulatory support: techniques of insertion. PMID- 8417699 TI - Chronic left ventricular support with a vented electric assist device. AB - Based on the success of the pneumatically powered HeartMate (Thermo Cardiosystems Inc, Woburn, MA) left ventricular assist device, researchers at the Texas Heart Institute have begun conducting clinical studies of the vented electric model in bridge-to-transplantation procedures. Like the pneumatic device, the electric device is implanted intraperitoneally, but the blood pump is powered through a percutaneous lead that is connected to two rechargeable batteries. The batteries are worn in a shoulder holster; thus, patients are not tethered to a control console and are fully mobile. Two patients are currently undergoing left ventricular support with the vented electric HeartMate (duration of support, 315 days and 112 days) as they await cardiac transplantation, and they seem to be medically fit for outpatient care and follow-up. This technology may eventually allow selected patients to receive long-term support as a substitute for cardiac transplantation. PMID- 8417700 TI - Univentricular and biventricular Thoratec VAD support as a bridge to transplantation. AB - As of October 1991, the Thoratec ventricular assist device (VAD) system has been used in 154 transplant candidates who were in imminent risk of dying before donor heart procurement at 39 medical centers in 10 countries. The VAD system consists of a prosthetic ventricle with a 65-mL pumping chamber made from Thoratec's BPS 215M polyurethane, cannulas for atrial or ventricular inflow and arterial outflow connections, and a pneumatic drive console. The devices can be used for partial or complete support of the pulmonary, systemic, or both circulations. In all patients (average age, 42 years; range, 11 to 64 years), the pumps were placed in a paracorporeal position on the anterior abdominal wall and connected to the heart and great vessels by cannulas crossing the chest wall. Biventricular support was used in 120 patients (78%) and isolated left VADs were used in 34. Average flow rate was 5.0 +/- 0.9 L/min for the left VAD and 4.3 +/- 0.8 L/min for the right VAD. The most frequent complications were bleeding (42% incidence, 7% mortality) and infection (36% incidence, 8% mortality). Ninety-eight patients (65%) recovered sufficiently to undergo heart transplantation after 8 hours to 226 days of support (average, 17.5 days), and 3 are waiting on VADs for transplantation. Eighty-two patients who received transplants have been discharged. This is an 84% early post-transplantation survival and a 54% overall survival. The actuarial survival 1 year after transplantation is 82%, comparable with that of conventional heart transplantation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417701 TI - Use of the Symbion biventricular assist device in bridging to transplantation. AB - From 1988 to 1991 13 patients received Symbion biventricular assist devices in attempts to bridge them to cardiac transplantation. All 7 of those who had cardiac transplants survived to hospital discharge. One death occurred 60 days after transplantation because of rejection. All other patients who received transplants are surviving. Implant times in this group varied from 10 to 164 days (mean, 55 days). There were two embolic neurologic events and two significant infections, and 2 of the survivors were dialyzed for reversible renal failure before transplantation. Of those who died on device support, 3 presented on centrifugal pump support. The three other deaths were caused by graft rejection, multiple organ failure, and multiple peripheral emboli. Biventricular assist devices optimally provide cardiac outputs of 4 to 5 L/min, can be quickly inserted often without requiring cardiopulmonary bypass, are easily explanted, and seem best suited for patients weighing less than 80 kg. PMID- 8417702 TI - Nursing care of the patient on mechanical circulatory support. PMID- 8417703 TI - Ventricular assist devices and total artificial hearts: a combined registry experience. AB - Data submitted voluntarily to the combined registry for the past 6 years on the use of ventricular assist devices for postcardiotomy cardiogenic shock in 965 patients and for circulatory support in conjunction with cardiac transplantation in 544 patients were analyzed. Of those patients whose ventricular function was expected to recover, approximately 45% were weaned from circulatory support and 25% were discharged from the hospital. Weaning status and hospital discharge were not different regardless of the type of original operative procedure or the pump design used. In the potential cardiac transplant group, 69% ultimately underwent transplantation and 66% were discharged from the hospital. The demographics (age and sex) of this group parallel those of patients undergoing isolated cardiac transplantation, and the 1- and 2-year survival estimates for patients requiring only univentricular support were equivalent to those of patients having isolated orthotopic cardiac transplantation. Ventricular assist devices are able to provide reasonable and safe circulatory support in both the postcardiotomy cardiogenic shock and the bridge-to-transplantation applications. PMID- 8417704 TI - Current expectations in dynamic cardiomyoplasty. AB - Dynamic cardiomyoplasty has been evaluated in the treatment of severe cardiomyopathies. This report outlines the results of this procedure in 21 patients with dilated or ischemic cardiomyopathy who were in New York Heart Association class III or IV before operation. There were no operative deaths. Patients were followed up for a mean of 17.6 months. Eight patients died during late follow-up, and actuarial survival rates were 73.2% at 1 year and 65.9% at 2 years of follow-up. Functional class improvement was documented in the surviving patients. Furthermore, significant improvement in left ventricular function was demonstrated by radioisotopic angiography and by heart catheterization for more than 2 years after the operation. These studies documented that left ventricular ejection fraction increased as a result of global improvement in regional wall motion. Absence of clinical and hemodynamic improvement after cardiomyoplasty seems to be related to muscle flap ischemic compromise, whereas the patient's condition before operation seems to influence the long-term outcome of cardiomyoplasty. PMID- 8417705 TI - Emergency resuscitation using portable extracorporeal membrane oxygenation. AB - Manual cardiopulmonary resuscitation is currently the standard treatment for cardiac arrest patients both in and out of the hospital. Accumulated experimental and anecdotal clinical evidence suggests enhanced survival in patients with extreme circulatory decompensation who have been emergently supported with portable cardiopulmonary bypass. Long-term survival is possible even when application is delayed, but early institution of support after cardiac arrest in selected patients offers the best survival advantages. In our hospital this has been achieved by training a team of in-house personnel to emergently prepare, apply, and temporarily manage cardiopulmonary bypass until personnel with greater specialty training arrive. Machinery needed to perform emergency cardiopulmonary bypass is currently available in all hospitals with open heart surgery programs. Simple support is often therapeutic but can also serve as a bridge to definitive diagnostic and other therapeutic procedures. Commercial units are becoming more biocompatible and easier to use, making both wider application and more prolonged support likely in the future. PMID- 8417706 TI - Pharmacological bridge to cardiac transplantation: current limitations. AB - Addition of intravenous enoximone to sympathomimetic agents permits a rapid and drastic improvement in the clinical and hemodynamical condition of patients in cardiogenic shock referred for a mechanical bridge to transplantation. The present experience, based on the management of 52 patients, permits us to point out the current limitations of this pharmacological bridge: the rate of sudden death, the incompleteness of the physical rehabilitation of the patients, and the vanishing effect of intravenous enoximone. PMID- 8417707 TI - How much do we need to know before approving a ventricular assist device? PMID- 8417708 TI - Warm blood cardioplegia: superior protection after acute myocardial ischemia. AB - Three myocardial protection techniques were studied in a canine model of acute myocardial ischemia with subsequent revascularization. Eighteen animals were randomly assigned to one of three treatment regimens: cold oxygenated crystalloid cardioplegia (CC), cold blood cardioplegia with modified reperfusate (CB), and continuous aerobic warm blood cardioplegia (WB) (n = 6 per group). Systemic hypothermic cardiopulmonary bypass (28 degrees C), antegrade arrest, and intermittent retrograde and antegrade delivery were used for the CC and CB groups. Systemic normothermic cardiopulmonary bypass, antegrade arrest, and continuous retrograde delivery were used for the WB group. Fifteen minutes of warm global ischemia was followed by occlusion of the left anterior descending coronary artery (15-minute duration) and simultaneous initiation of cardioplegic arrest (60-minute duration) to simulate clinical revascularization. After reperfusion, the animals were separated from cardiopulmonary bypass. Myocardial function, electrocardiogram, myocardial energetics, water content, histopathology, and defibrillation requirements were compared between groups. There was no significant difference in maximum elastance, myocardial oxygen consumption, myocardial edema, or histopathologic evidence of injury between groups. However, overall ventricular function, assessed by the slope of the preload recruitable stroke work relationship, was significantly better for the WB group (p = 0.04) (WB, 73 +/- 9; CB, 56 +/- 7; CC, 47 +/- 5). Diastolic function as assessed by the slope of the stress-strain relationship was significantly worse overall for the cold groups (p = 0.001) (WB, 20 +/- 2.2; CB, 39 +/- 1.3; CC, 37 +/- 3.1). Myocardial injury as assessed by ST segment elevation (millimeters) was less for the WB group (p = 0.03) (WB, 0.4 +/- 0.3; CB, 1.7 +/- 0.2; CC, 1.6 +/- 0.7). Countershocks necessary to restore sinus rhythm after cross-clamp removal were fewer in the WB group (p = 0.03) (WB, 0.8 +/- 0.3; CB, 4.0 +/- 1.2; CC, 5.5 +/- 1.5). In this model of acute global myocardial ischemia, continuous aerobic warm blood cardioplegia has important advantages over two widely used clinical hypothermic protection techniques. PMID- 8417709 TI - Total anomalous pulmonary venous connection in neonates and young infants: repair in the current era. AB - Total anomalous pulmonary venous connection has been one of the more challenging congenital heart defects in newborns and young infants despite improvements in surgical technique, cardiac anesthesia, neonatal myocardial preservation, and postoperative care. Since 1981, 30 patients with total anomalous pulmonary venous connection have undergone primary total correction. Mean age at operation was 28 +/- 6 days and mean weight, 3.3 +/- 0.7 kg. Essential features of the surgical approach in these small patients included early surgical intervention, profound hypothermia with total circulatory arrest, cardioplegic myocardial preservation, and a wide posterior anastomosis. Operative mortality was 13% +/- 6%. All four deaths were in patients having emergency operation within 24 hours of the surgical consult and requiring ventilator support preoperatively. The mean follow up is 47 +/- 7 months. There have been two late deaths, and the 7-year survival rate is 79% +/- 8%. There have been two reoperations, and 91% +/- 6% of the patients are reoperation free at 7 years. Only 1 of the 24 surviving patients is symptomatic. Growth in survivors is closely monitored. The height growth percentile is less than 5% in 15% +/- 8% of survivors and the weight growth percentile, less than 5% in 17 +/- 8%. During the past decade, with a consistent surgical approach to neonates and infants with total anomalous pulmonary venous connection, it has been possible to achieve low early mortality, low attrition, and excellent late results. PMID- 8417710 TI - RBRVS and cardiothoracic surgery. PMID- 8417711 TI - Effects of University of Wisconsin solution on endothelium-dependent coronary artery relaxation in the rat. AB - University of Wisconsin (UW) solution has been reported to enhance myocardial preservation in heart transplantation. To evaluate the effects of UW solution on coronary artery endothelial function, we designed experiments to compare UW solution with a standard crystalloid hyperkalemic cardioplegic solution (CHCS). Isolated rat hearts were studied in a modified Langendorff apparatus for coronary endothelial function. Groups 1 and 2 were perfused with 4 degrees C CHCS (24 mmol/L of KCl) and UW solution, respectively, for 10 minutes at a pressure of 80 cm H2O, whereas group 3 underwent warm ischemia for 10 minutes. Groups 4 and 5 were perfused with and stored for 4 hours in cold (4 degrees C) CHCS and UW solution, respectively. Group 6 underwent 4 hours of topical cooling (4 degrees C) without any cardioplegic perfusion. All groups had 6 hearts each. Endothelium dependent relaxation and endothelium-independent relaxation of the coronary arteries were tested by infusing 5-hydroxytryptamine (5HT) (10(-6) mol/L) and sodium nitroprusside (10(-5) mol/L), respectively, before and after perfusion with and storage in one of the two cardioplegic solutions. The coronary vasodilatation induced by 5HT and sodium nitroprusside was not altered in hearts perfused with (group 1) or perfused with and stored in CHCS (group 4). Coronary flow increase after 5HT infusion was significantly decreased in hearts perfused with (group 2) (before, 35% +/- 10%; after, 13% +/- 10%; p < 0.01) or perfused with and stored in UW solution (group 5) (before, 34% +/- 5%; after, -5% +/- 12%), indicating severe endothelial dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417712 TI - Cerebral metabolism and circulatory arrest: effects of duration and strategies for protection. AB - Hypothermic total circulatory arrest (CA) is commonly used to facilitate repair of complex congenital heart defects. However, the "safe" period of CA remains to be defined. Extended periods of hypothermic total circulatory arrest may impair cerebral metabolism and cause ischemic brain injury. This study defines the relationship between increasing durations of CA at 18 degrees C and cerebral metabolism, and examines the protective value of topical cooling of the head or continuous "trickle" flow (5 to 10 mL.kg-1.min-1). Thirty-three 1-week-old piglets were randomized to six experimental groups: control; 15, 30, or 60 minutes of CA; 60 minutes of CA with topical cooling of the head; and 60 minutes of trickle flow. Animals were placed on cardiopulmonary bypass (CPB) at 100 mL.kg 1.min-1 and cooled to 18 degrees C. After the experimental period of CA or trickle flow (or 60 minutes of CPB at normal flow for the control group), animals were rewarmed to 37 degrees C and weaned from CPB. Data were obtained before and immediately after CPB at 37 degrees C, and before and immediately after the experimental period at 18 degrees C. Parameters measured included cerebral blood flow by xenon 133 clearance, arterial and sagittal sinus blood gases, and cerebral metabolism. Hypothermic total circulatory arrest caused an impairment of cerebral metabolism that was directly proportional to CA duration (r2 = 0.73; p = 0.0001), and recovery of metabolic function after 60 minutes of CA improved more than 50% if the head was packed in ice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417713 TI - Improved multiorgan function after prolonged univentricular support. AB - Eleven cardiac transplant candidates (all male; mean age, 43.3 years) with multiorgan (hepatic, pulmonary, and/or renal) dysfunction were sustained for prolonged periods (> 30 days) with the HeartMate (Thermo Cardiosystems, Inc, Woburn, MA) left ventricular assist device. We evaluated the effect of extended support on end-organ recovery and on the ultimate outcome of cardiac transplantation. In addition to cardiac failure, 9 patients had hepatic dysfunction, 8 had pulmonary dysfunction, and 6 had renal dysfunction (4 of whom required hemodialysis before left ventricular assist device support). Mean duration of support was 115 days (range, 31 to 233 days). All patients underwent successful transplantation; 10 of these patients survived a mean of 24 months. One patient, who had required hemodialysis and ventilatory support during and after support, experienced progressive multiorgan failure and died 7 weeks after transplantation. Two late deaths after transplantation were unrelated to the device. Overall, patients experienced improvement in cardiac functional class status, and most participated in cardiac rehabilitation programs before transplantation. During left ventricular assist device support, hepatic function returned to normal in 8 patients, pulmonary function recovered in 7, and renal function returned to normal in 4. One patient who required hemodialysis underwent renal transplantation after cardiac transplantation and had complete recovery of renal function. In the current era of donor shortages, gravely ill patients can benefit from a strategy of prolonged left ventricular assist device support. This strategy has proved safe, has allowed for reversal of multiorgan dysfunction, and has produced healthier transplant candidates. PMID- 8417714 TI - Component analysis of bilateral anterior cardiomyoplasty. AB - This experiment was designed to analyze the mechanism of ventricular augmentation generated with bilateral anterior cardiomyoplasty by comparing it in an acute setting with its components, the left anterior cardiomyoplasty and the right anterior cardiomyoplasty. Hemodynamic variables were measured in 8 dogs before and after each flap was positioned around the heart. Stimulation was achieved by R-wave synchronous latissimus burst pacing at a ratio of 1:4, an amplitude of 5 V, a frequency of 33 Hz, and a duration of 30% of the R-R interval. Hemodynamic changes were again recorded during latissimus stimulation. Construction of the bilateral anterior wrap (static cardiomyoplasty) caused some depression of baseline hemodynamic function, which was greater than that caused by either the static right or left anterior cardiomyoplasty. With stimulation of the muscles (dynamic cardiomyoplasty), the bilateral wrap caused significant biventricular augmentation. Evaluation of the components of the bilateral wrap demonstrated that dynamic right anterior cardiomyoplasty also provided significant biventricular augmentation, but the dynamic left anterior cardiomyoplasty augmented only right-sided variables. The mechanism of biventricular compression by the bilateral procedure is due mostly to the right wrap. The right anterior cardiomyoplasty may provide significant biventricular compression for treatment of heart failure, without the complexity associated with bilateral anterior cardiomyoplasty. PMID- 8417715 TI - Skeletal muscle ventricles: left ventricular apex to aorta configuration. AB - Skeletal muscle ventricles (SMVs) were constructed from the latissimus dorsi muscle in 6 dogs. After 3 weeks of vascular delay followed by 6 weeks of 2-Hz continuous electrical conditioning, a valved conduit was placed between the left ventricular apex and the SMV and a second valved conduit, between the SMV and the aorta. The SMV was stimulated to contract during diastole at a 1:2 ratio with the heart. The SMV pumped 47% of the systemic blood flow initially (0.73 +/- 0.23 versus 1.54 +/- 0.42 L/min) and 40% after 3 hours. Skeletal muscle ventricle stimulation resulted in a 58% increase in mean diastolic pressure initially (52 +/- 9 to 82 +/- 11 mm Hg; p < 0.05) and a 73% increase (45 +/- 7 to 78 +/- 8 mm Hg) after 3 hours of continuous pumping. This was associated with a 68% increase in the endocardial viability ratio initially and a 63% increase at 3 hours. The systolic tension-time index decreased by 26% initially and 25% at 3 hours. This study indicates that the SMV configuration of left ventricular apex to aorta may be particularly suitable for left ventricular assist. PMID- 8417716 TI - Prophylactic digitalization fails to control dysrhythmia in thoracic esophageal operations. AB - A prospective, controlled, randomized study of 80 patients undergoing esophageal operations was undertaken, in which one group of patients was given digoxin and the other was not. The incidence of cardiac dysrhythmia was compared in each group. Twenty-six patients underwent operation for benign disease. Equal numbers were digitalized or not and no dysrhythmias occurred. Fifty-four patients underwent operation for malignant disease. Of 26 in the group digitalized, 12 suffered dysrhythmia (46%). Of 28 not digitalized, 9 suffered dysrhythmia (32%). Overall, 39% of patients with malignant disease suffered a dysrhythmia compared with none with benign disease (p < 0.002 by chi 2). PMID- 8417717 TI - Benefits of posttransplantation monitoring of interleukin 6 in lung transplantation. AB - To determine the predictive diagnostic value of interleukin 6 (IL-6) monitoring in lung and heart-lung transplants, we measured posttransplantation serum IL-6 levels in 17 adult lung or heart-lung transplant recipients. Posttransplantation IL-6 elevation patterns were classified into 4 groups: serum IL-6 level remained negative throughout the monitoring period (group 1; n = 1; 6%); several sharp spikes with normal baseline (group 2; n = 9; 53%); persistently high level of serum IL-6 (group 3; n = 3; 18%); and several sharp spikes of serum IL-6 elevation with abnormally high baseline (group 4; n = 4; 24%). One patient without an elevation of IL-6 (group 1) did not experience any episodes of rejection or infection. Nine patients in group 2 had 19 IL-6 spikes, 13 of which were associated with histopathologically or clinically diagnosed rejection, 3 with acute bronchitis, and 1 with diffuse alveolar damage. Three patients in group 3 had persistent infections including cytomegalovirus infection, toxic megacolon, and repeated bacterial infection during the monitoring period, and 4 in group 4 died within 3 months after transplantation. From this study it appears that a spiked elevation of IL-6 could have a predictive value in diagnosing rejection, and persistently high levels of IL-6 indicate the presence of infection. Thus, IL-6 monitoring is beneficial for lung transplant recipients. PMID- 8417718 TI - Upper gastrointestinal dysmotility in heart-lung transplant recipients. AB - Recipient pneumonectomy and the necessity for meticulous hemostasis in heart-lung transplantation can result in injury to the vagus nerves as they course through the posterior mediastinum, with consequent delay in gastric emptying. This has been reported to lead to chronic aspiration and associated pulmonary sequelae. To study the association between delayed gastric emptying, bronchiectasis, and bronchiolitis obliterans after heart-lung transplantation, we performed esophageal manometry, 24-hour pH monitoring, and radioisotopic gastric emptying in 10 patients who underwent heart-lung transplantation. Three patients had grossly delayed liquid and solid emptying that was compatible with complete vagotomy. Six other patients had delayed liquid but normal solid emptying--an unexplained finding that is the reverse of what one would expect from vagal injury. Two of these 9 patients had esophageal dysmotility, but none demonstrated gastroesophageal reflux. One remaining patient had faster than normal gastric emptying for both solids and liquids. Of the 10, 2 patients have radiologic changes of bronchiectasis and 3 have biopsy evidence of obliterative bronchiolitis. There is no relationship between these sequelae and the occurrence of esophageal dysmotility, gastroesophageal reflux, or vagotomy. We conclude that gastric emptying abnormalities can occur after heart-lung transplantation, but such abnormalities are not associated with gastroesophageal reflux and the development of pulmonary sequelae, as previously reported. PMID- 8417719 TI - Surgical management of thrombotic disc valve. AB - Thrombotic obstruction, a rare but often fatal complication of cardiac valve prostheses, appears to occur more frequently in tilting-disc valves than in other valve designs. Its diagnosis and surgical treatment remain a challenge. Ten consecutive patients who had thrombosis of a tilting-disc valve prosthesis were treated in Chang Gung Memorial Hospital from November 1982 to August 1990. Preoperative clinical features, including exertional dyspnea, new murmur, and absence of a metallic click from the prosthetic valve, occurred in all of the patients. Symptoms were present for 1 week or more before reoperation in 70% of the patients; nevertheless, many patients were referred only after acute exacerbation of heart failure and development of pulmonary edema. Echocardiography confirmed prosthetic valve malfunction in 90% of the patients. One unconfirmed case was later documented by cardiac catheterization. Anticoagulant therapy was in the therapeutic range for only half of the patients at the time of admission. Prompt reoperation was performed for thrombectomy (8 patients, all survived) or valve replacement (2 patients, one death). Long-term outcome was satisfactory in all survivors with a mean follow-up of 31.6 months. These findings emphasize the importance of considering the diagnosis of thrombosis in patients with mechanical heart valve prostheses who are first seen with nonspecific symptoms and minor changes of their physical findings. The diagnosis could be easily made by echocardiography. Thrombectomy is an easy, fast, and safe procedure, especially for these critically ill patients. PMID- 8417720 TI - Perceived controllability and the development of posttraumatic stress disorder (PTSD) in crime victims. AB - This study evaluated the association between perception of controllability and the development of posttraumatic stress disorder (PTSD) following criminal assault. Factor analysis of a perceived controllability scale revealed three factors; perceived controllability felt during the assault, expected controllability over future assaults, and perceived controllability over aversive events more generally. Only the latter factor was associated with PTSD symptom severity. The hypothesis that perceived controllability would be negatively associated with assault severity was partially supported. Further analyses showed that the association between controllability and PTSD was not mediated or moderated by assault severity measures. The role of perceived controllability in the development of PTSD is discussed. PMID- 8417721 TI - Self-reports of panic attacks and manifest anxiety in adolescents. AB - A recent paper by Nelles and Barlow (1988; Clinical Psychology Review, 8, 359 372) provided the rationale for an investigation of panic attacks in adolescents. A panic attack questionnaire and the Revised Children's Manifest Anxiety Scale were administered to an unselected sample of Australian adolescents. Of 534 adolescents, 42.9% reported having at least one panic attack. Other data are reported on the characteristics of panic attacks, life interference and avoidance behaviour. Panickers reported significantly higher anxiety levels than nonpanickers. Differences between the findings of Australian and American samples were noted and directions for future research were identified. Several methodological issues were also discussed including the reliability and validity of self-report data on panic attacks. PMID- 8417722 TI - On the termination of panic attacks. A reply to Ley. PMID- 8417723 TI - Do patients suffering from obsessions alone differ from other obsessive compulsives? AB - The aim of this study was to determine whether a group of 48 patients with obsessions and compulsions differed from a group of 26 patients with obsessions alone, on the basis of demographic variables, variables relating to obsessive compulsive neurosis, psychological variables and treatment outcome. It was found that they differed significantly in the following areas: marital status, level of education, age at onset of complaints, psychoactive medication taken when admitted for treatment, severity of obsessive-compulsive complaints, depression and intelligence. No difference was found as far as treatment outcome was concerned. Patients suffering from obsessions alone, would appear to form a distinct sub-group within the group of obsessive-compulsive patients as a whole. PMID- 8417724 TI - Negative recall bias and neuroticism: state vs trait effects. AB - Incidental recall of positive and negative trait adjectives was assessed in high and low N (neuroticism) Ss in whom depressed or neutral mood had been induced. Results indicated that the negative recall bias is an interactive function of trait vulnerability (as reflected by N scores) and current mood state. In the depressed mood condition, there was a trend for high N to be associated with relatively better recall of negative material, as expected. However, in neutral mood, high levels of N predicted relatively poorer recall of negative information. This unexpected finding was discussed in relation to previous research into mood-incongruent recall effects and sex differences. PMID- 8417725 TI - Performance demand and sexual arousal in women. AB - Up to now, no experimental studies have inquired into the possible role of performance demand in female sexuality. The objective of this study is to investigate the effects of performance demand on sexual arousal in functional women, using explicit instructions. Forty-eight female subjects were asked to respond with maximum sexual arousal within 2 min, both during sexual fantasy and while watching an erotic film excerpt. Photoplethysmographic pulse amplitude was continuously recorded and self-report ratings of sexual arousal and affective reactions were collected after each erotic condition. Subjects were also instructed to continuously indicate their subjective sexual excitement during the conditions by means of a lever. Performance demand resulted in higher genital responses and was most effective in the fantasy condition. These results corroborate the findings for functional men. Although for both measures of subjective experience of sexual arousal performance demand yielded significantly higher ratings, this was conditioned by the order in which subjects were exposed to conditions. Women who masturbate with a mean frequency of 4 times a month reported higher subjective sexual arousal during performance demand conditions as compared with women who masturbate substantially more or who do not masturbate. Genital vasocogestion did not differ between masturbation-groups. Continuous subjective monitoring yielded lower correlations with vasocongestion than discrete ratings of sexual arousal and appears to be more sensitive to order effects. PMID- 8417726 TI - Suicide, chemical abuse, and panic attacks: a preliminary report. AB - Seventy-two chemical abuse patients who either met DSM-III-R criteria for panic disorder (PD), experienced infrequent panic attacks (IP), or did not experience panic attacks in the past year (NP) were compared on several dimensions. PD subjects were more likely to be female and to have attempted suicide. Patients who had attempted suicide, when compared to non-suicide attempters, were more likely to be classified as having PD, and be non-married. IP and NP groups did not differ on any of the relevant variables. The similarities of these findings to those obtained with PD and chemical abuse patients are discussed. PMID- 8417727 TI - Familial resemblances in disgust sensitivity and animal phobias. AB - This paper reports the results of two studies, both of which suggest there are close intrafamilial resemblances between parental disgust sensitivity and various measures of offspring animal phobia. In Study 1, a multiple regression analysis identified parental disgust sensitivity as a primary predictor of offspring animal fear in general (as measured by the animal phobia sub-scale of the Fear Survey Schedule), and offspring spider fear in particular. Study 2 indicated that parental disgust sensitivity was only related to offsprings' fear of animals known to be associated with the disgust reaction. These intrafamilial resemblances are discussed in relation to genetic and cultural transmission of the disgust reaction. PMID- 8417728 TI - A comparison of three worry questionnaires. AB - This paper describes a study comparing three worry questionnaires; The Worry Domains Questionnaire (WDQ), The Penn State Worry Questionnaire (PSWQ), and The Student Worry Scale (SWS). The results suggested that (i) scores on two out of the three questionnaires exhibited sources of variance that were independent of trait anxiety, (ii) content-based questionnaires (the WDQ and SWS) appeared to capture features of task-oriented constructive worrying, whereas the PSWQ did not, and (iii) all three questionnaires indicated that worriers were characterized by an information-seeking, monitoring cognitive style and a tendency to indulge in avoidance coping behaviours. The implications of these findings for the development of a clinically useful diagnostic instrument are discussed. PMID- 8417729 TI - Emotionality and interference with color-naming in anxiety. AB - The source of interference with color-naming emotional words in clinical anxiety was investigated using word sets that were varied in valence and in their judged relationship to the concerns of anxious patients. Results showed that neither valence nor general emotionality was of critical importance in predicting extent of interference. In contrast, words that were judged to be highly related to likely concerns or relevant threats caused more interference than those which were not, irrespective of their positive or negative valence. These results suggest the need to modify earlier formulations of emotional Stroop effects, and may help to explain the variable results obtained in previous investigations. PMID- 8417730 TI - Assessment of pain-related cognitions in chronic pain patients. AB - The construction of the Pain-Related Self Statements Scale (PRSS) and the Pain Related Control Scale (PRCS) is described. The PRSS assesses situation-specific aspects of patients' cognitive coping with pain, while the PRCS measures general attitudes towards pain. The reliability and validity of these scales were determined in a sample of 120 chronic pain patients suffering from various rheumatic disorders, 213 patients who suffered from chronic back pain, 44 patients with temporomandibular pain and dysfunction and 38 healthy controls. The analysis of the PRSS yielded two scales termed 'Catastrophizing' and 'Coping'; the PRSC consists of the subscales 'Helplessness' and 'Resourcefulness'. All four subscales were demonstrated to be valid and sensitive to change, and they are closely related to pain intensity and interference from pain experiences. PMID- 8417731 TI - Preattentive processing, preparedness and phobias: effects of instruction on conditioned electrodermal responses to masked and non-masked fear-relevant stimuli. AB - We hypothesized that autonomic responses conditioned to fear-relevant stimuli, in contrast to responses conditioned to neutral stimuli, can be elicited after only an automatic, non-conscious analysis of the stimulus. Consequently, they may be expected to be insensitive to verbal instructions. Normal subjects were conditioned to either fear-relevant stimuli (snakes or spiders) or neutral stimuli (flowers or mushrooms) in a differential conditioning paradigm with shock as the unconditioned stimulus. In a subsequent extinction series, half of the subjects were shown the conditioned stimuli under masking conditions preventing their conscious recognition, whereas the other half were exposed to non-masked stimuli. Then half of the subjects in each of the masking conditions were verbally instructed that no more shocks would be delivered and then the extinction trials followed. Consistent with our hypothesis, differential responses to the fear-relevant CSs+ and CSs- remained unaffected by both masking and instruction, whereas differential responding to neutral stimuli was wiped out by the masking procedure and the verbal instruction. PMID- 8417732 TI - The Staats-Heiby theory of depression: the role of event frequency and affect reevaluated. AB - This paper reports an attempt to replicate and extend a study by Rose and Staats (Behavior Research and Therapy, 26, 489-494, 1988) and reviews their attempts to find support for two subtypes of depression as predicted by the Staats-Heiby theory of unipolar depression. Due to methodological and analytical problems in the work by Rose and Staats (1988) it was felt that the conclusions they drew were unjustified. In this study 160 university students completed the Mood Related subscales of the Pleasant (PES-MR) and Unpleasant Event Schedules (UES MR) and the Beck Depression Inventory. Overall, the contention that different subtypes of depression could be differentiated on the basis of the interaction between frequency of events and the affective value of events was not supported. However, there was some support for the role of pleasant event frequency, unpleasant event frequency, and event unpleasantness in depression. The results indicate that the relationship between event frequency and hedonic strength differs between the PES-MR and UES-MR. It is argued that the research methodology and analyses used could not adequately assess the hypotheses generated from the Staats-Heiby model. PMID- 8417733 TI - Long-term sulfamethoxazole-trimethoprim in Wegener's granulomatosis. AB - Since 1985, a number of reports have highlighted the effectiveness of a short course of sulfamethoxazole-trimethoprim as an adjuvant to immunosuppressive drugs in the treatment of Wegener's granulomatosis. We report our experience with long term sulfamethoxazole-trimethoprim therapy in patients with Wegener's granulomatosis. We describe recent advances in the pathogenesis of Wegener's granulomatosis, suggest a complementary mechanism of action of sulfamethoxazole trimethoprim, and advocate long-term sulfamethoxazole-trimethoprim therapy as a treatment option in Wegener's granulomatosis. PMID- 8417734 TI - Autosomal dominant sensorineural hearing loss. Further temporal bone findings. PMID- 8417735 TI - Unifocal Langerhans' cell histiocytosis (eosinophilic granuloma) of the petrous apex. AB - Eosinophilic granuloma is a localized form of histiocytosis X, or Langerhans' cell histiocytosis, a benign lesion of proliferating Langerhans' cells. It is the least severe of the histiocytosis syndromes, and is characterized by lytic lesions of one or more bones. Temporal bone lesions usually occur in association with multifocal disease; however, isolated lesions may occur in either the mastoid bone alone, or in the entire temporal bone, without disease elsewhere in the body. We present the first case (to our knowledge) of eosinophilic granuloma, or unifocal Langerhans' cell histiocytosis, limited to the petrous apex. The patient, an 8-year-old girl, presented with facial nerve paralysis. Because of delay in definitive diagnosis, the disease extended beyond its original boundaries and resulted in complete destruction of the temporal bone. We will also review Langerhans' cell histiocytosis, with attention to its involvement of the temporal bone. PMID- 8417736 TI - Burkitt's lymphoma of the parapharyngeal space. AB - Burkitt's lymphoma is quite rare outside of Africa. It is even more uncommon for this nonendemic form of the disease to present in the head and neck region. To our knowledge, nonendemic Burkitt's lymphoma has not been previously reported arising from the parapharyngeal space. We review the case of a 10-year-old boy who presented with an asymptomatic mass at the angle of the mandible several weeks after blunt trauma to that area. We discuss the evaluation of this mass, including the results of computed tomography and magnetic resonance imaging, along with the findings of histopathologic and cytogenetic studies. We also discuss the differential diagnosis of parapharyngeal lesions, paying particular attention to the workup and management of Burkitt's lymphoma. Since Burkitt's lymphoma has the highest growth rate of any tumor in man, rapid diagnosis and immediate treatment are important and likely to improve outcome. PMID- 8417737 TI - Head and neck cancer. PMID- 8417738 TI - Imaging quiz case. Noninvasive Aspergillus sinusitis. PMID- 8417739 TI - Calvarial bone graft harvest. Techniques, considerations, and morbidity. AB - The importance of calvarial bone grafting in craniomaxillofacial trauma and facial reconstructive surgery is now widely recognized. Numerous harvesting techniques have evolved to optimize the desired thickness, size, shape, and curvature for a particular reconstructive need. At the same time, donor site selection and morbidity must be considered. This report includes a discussion of several currently utilized calvarial bone graft harvesting techniques, with emphasis on minimizing complications and morbidity. Choice of optimal harvesting technique and donor site for specific reconstructive situations will likewise be discussed. Finally, the specific morbidity in our multi-institutional calvarial bone graft harvest series of 121 patients and over 350 grafts will be reviewed. PMID- 8417740 TI - Irradiated cartilage grafts in the nose. A preliminary report. AB - Many nasal deformities require some form of graft or implant material for complete correction. Various well-recognized disadvantages are associated with currently used autografts, allografts, and alloplastic materials, thereby maintaining a continued search for the ideal nasal graft. Irradiated homograft costal cartilage is an allograft that has been used with variable success in the head and neck region. Isolated reports have suggested favorable results with irradiated homograft costal cartilage implantation in the nose. This study details the findings of 306 irradiated homograft costal cartilage grafts that were used in 122 nasal augmentation procedures. PMID- 8417741 TI - The human auricular chondrocyte. Responses to growth factors. AB - The first human auricular (elastic cartilage) chondrocyte cell culture model is presented. These chondrocytes, harvested from fresh human cadaver auricular cartilage, are easily grown in culture. They produce type II collagen and abundant alkaline phosphatase. Like growth plate chondrocytes, human auricular chondrocytes respond mitogenically to both transforming growth factor-beta and basic fibroblast growth factor by increasing proliferation twofold. The growth factors exert a synergistic effect on thymidine uptake in this model, with maximal stimulation occurring at the dose combination of basic fibroblast growth factor, 10 ng/mL, and transforming growth factor-beta, 3 ng/mL. Human nasal septal (hyaline cartilage) chondrocytes do not increase proliferation in response to these two growth factors. Human auricular chondrocytes also increase matrix production in response to transforming growth factor-beta, as indicated by increased proteoglycan production and increased collagen synthesis. The increased matrix production combined with increased proliferative rate elicited with transforming growth factor-beta and basic fibroblast growth factor indicates that human elastic cartilage chondrocytes harvested from the external ear are better suited for in vitro cartilage implant growth than human hyaline cartilage chondrocytes. Further biochemical and molecular biological characterization of the human auricular chondrocyte cell culture model is currently under way. Additional work is also under way to generate multilayer culture growth on absorbable frameworks, in the pursuit of the long-term goal of in vitro production of autogenous cartilage implants. PMID- 8417742 TI - Comparison of balance assessment by sway magnetometry and force platforms. AB - Sway magnetometry, which monitors body movement at the hips, was compared with two commercial force platforms to determine its sensitivity in identifying differences in stability between eyes opened and eyes closed. Sway path length and area were examined. All devices showed an ability to distinguish between stability with eyes opened and eyes closed on repeated testing. The magnetometry system using path length analysis was most sensitive and was able to detect an increase in sway with eyes closed compared with eyes open for individual tests with 97% confidence (mean [+/- SD] path length Romberg Coefficient, .76 +/- .13). Standing on a rubber base caused destabilization of posture. Sway magnetometry is seen as a potentially useful tool in quantification of body sway to aid the clinician in assessment of balance in the dizzy patient. PMID- 8417743 TI - Ear-nose-throat abnormalities in the CHARGE association. AB - A comprehensive evaluation of the otolaryngological abnormalities in 50 patients with colobomata, heart defect, atresia of the choanae, retarded growth or development, genital hypoplasia, and ear anomalies or deafness (CHARGE) was performed. All the patients had ear abnormalities; 96% (48/50) had malformed pinnae, and 54% (27/50) had facial nerve palsies. Only 8% (4/50) had normal hearing, the commonest hearing defect being severe conductive or mixed loss. Eighty-four percent (42/50) of computed tomographic scans of the temporal bone were abnormal, the characteristic abnormality being the combination of a hypoplastic incus and absent semicircular canals. Eighty-six percent (43/50) of patients had upper airway abnormalities. Posterior choanal abnormalities occurred in 56% (28/50), and 42% (21/50) had retrognathia leading to intubation difficulties. Laryngotracheal abnormalities occurred in 38% (19/50), and 14% (7/50) required tracheostomies. Careful upper airway assessment is essential to avoid potentially lethal complications such as aspiration. PMID- 8417744 TI - Combined electrical and acoustical stimulation using a bimodal prosthesis. AB - A new device incorporating a cochlear implant speech processor and a speech processing hearing aid for the unimplanted ear has been designed and tested with four severely hearing-impaired patients. The aim of the device is to provide a more acceptable and effective combination of electrical and acoustic signals to the two ears. When used monaurally, and binaurally in conjunction with the cochlear implant, the speech-processing hearing aid mean scores for open-set sentences, words, and consonants were as good as or better than the mean scores for the patients' own conventional hearing aids. Some patients improved much more than did others. Although not conclusive, these results are encouraging, especially as they were achieved with a laboratory prototype that did not allow the patients to become accustomed to the processor in everyday situations. PMID- 8417745 TI - Impact of bilateral neck dissection on recovery following supraglottic laryngectomy. AB - Previously reported data from our institution has led us to perform bilateral neck dissections for therapeutic as well as staging advantages for horizontal supraglottic laryngectomies. Concern over the possibility of increased morbidity associated with simultaneous bilateral neck dissection prompted this retrospective review of patients with supraglottic laryngectomy who were treated with either unilateral (46 patients) or bilateral (23 patients) neck dissection. No significant differences were found in morbidity when patients were evaluated for transfusion rate, cervical wound drainage, need for tracheotomy, oral diet, or duration of hospitalization. Significant differences were noted in surgical operating time, eg, it took 100 minutes longer to perform bilateral dissections, and slight increases were noted in estimated blood loss and fluids given intravenously. No significant differences were noted in the percentage or type of postoperative complications. It seems that bilateral neck dissection in conjunction with supraglottic laryngectomy does not increase postoperative surgical morbidity and may actually avoid complications associated with postoperative radiation therapy in patients with supraglottic laryngectomy. PMID- 8417746 TI - Analyses of distant metastases in squamous cell carcinoma of the head and neck and lesions above the clavicle at autopsy. AB - The frequency of distant metastases was studied in 112 patients who had squamous cell carcinoma of the head and neck. All of these patients died of the tumor and underwent postmortem examinations. Forty-one (37%) of the 112 patients had distant metastases at the time of death. There was a significant correlation between development of distant metastases and the presence of cervical nodes. Of the patients who had distant metastases, 63% had evidence of tumor in the cervical nodes, whereas 37% of these patients were free of disease in the neck. Patients with residual or recurrent tumors in the cervical nodes had a high risk of distant metastases. PMID- 8417747 TI - Natural killer cell activity in laryngeal carcinoma. AB - We studied the relationship of natural killer cell activity from peripheral blood mononuclear cells with the clinical and pathologic stage of disease in 23 male patients with previously untreated carcinoma of the larynx and 22 healthy male control subjects. Levels of natural killer cell activity against K-562 target cells were similar in control subjects and patients, regardless of stage, tumor size, and clinical cervical adenopathies. Natural killer cell activity, however, was significantly decreased in patients with pathologic cervical lymph node involvement. The number of natural killer cells, as estimated by CD16 and CD56 monoclonal antibodies, was similar in all groups of subjects. We conclude that in patients with laryngeal carcinoma, there is a correlation between deficient natural killer cell activity and nodal metastases, which may represent a prognostic indicator in these patients. PMID- 8417748 TI - Tracheal view of vocal fold vibration in excised canine larynxes. AB - The mucosal upheaval where the mucosal wave starts and propagates upward appears on the lower surface of the vocal fold during vibration. We investigated the location of the mucosal upheaval in response to variations in vocal fold tension and mean air flow rate. Twelve excised canine larynxes were used in the experiment. The lower surfaces of the vocal folds were marked with india ink. After cinematography was performed, a small cut wound was made at the mark. This wound served to relate cinematographic findings with the histologically observed position of the mark. The larynx was then fixed, sectioned in the frontal plane, and stained for histologic examination. Horizontal movements of the free edge, mucosal wave, mucosal upheaval, and marks were analyzed. Under a fixed vocal fold tension, the mucosal upheaval appeared more laterally to a limited extent as mean air flow rate increased, but its location on the vocal fold mucosa did not change from its position before the increase of mean air flow rate. The mucosal upheaval appeared more medially when vocal fold tension increased. The position of the mucosal upheaval actually changed medially compared with its original position before the tension increase. Histologic examination indicated that the mucosal upheaval arose on the lower surface of the vocal fold between the free edge and the area where the muscular layer comes in close proximity to the epithelial layer. PMID- 8417749 TI - Treatment of laryngotracheal stenosis with anterior and posterior cartilage grafts. A report of 41 children. AB - Forty-one tracheotomy-dependent children with moderate and severe laryngotracheal stenosis (Cotton grades III and IV) underwent laryngotracheal reconstruction with simultaneous use of costal cartilage grafts in the anterior and posterior subglottis and posterior glottis. The indications for this procedure are the presence of stenosis in the posterior part of the glottis, subglottis, or both in combination with anterior subglottic or upper tracheal narrowing. Other indications are the presence of circumferential subglottic stenosis, bilateral subglottic shelves, or total subglottic obstruction. The complications of this procedure included pneumothorax (one patient) and granulation tissue formation (five patients). There were two deaths unrelated to the reconstructive procedure. This technique offers an excellent opportunity for repair of laryngotracheal stenosis with a better than 90% tracheotomy decannulation rate after a single procedure within a short period after removal of the stent. PMID- 8417750 TI - Surface contour three-dimensional imaging in congenital aural atresia. AB - Sixty-five patients with congenital aural atresia-stenosis had three-dimensional reconstructions of their high-resolution computed tomographic scans. Surface anatomy of the temporal bone was readily demonstrated, including its relation to the temporomandibular joint. Three important findings were noted. (1) Contrary to popular belief, the condyle of the mandible does not rest against the anterior face of the mastoid bone. (2) A bony cleft or groove is often in the lateral temporal bone through which the facial nerve may exit. (3) Duplications of bony structures attached to or part of the temporal bone are clearly defined. The information gained from the routine use of three-dimensional imaging of the computed tomographic scan alerts us to potential intraoperative problems that may otherwise escape our scrutiny, particularly if only two-dimensional computed tomographic scanning is done. PMID- 8417751 TI - Mast cells and biogenic amines in radiation-induced pulmonary fibrosis. AB - Sprague-Dawley rats were exposed to a single X-ray dose of 30 Gy over the lungs and examined at 1-wk intervals during the following 3 to 8 wk. Mast cells were counted after specific staining with toluidine blue at a low pH and the mast-cell amines, histamine (Hi) and serotonin (5-HT), were measured using high-performance liquid chromatography. Irradiation induced pneumonitis followed by pulmonary mast cell hyperplasia and progressive fibrosis 4 to 8 wk after irradiation. By week 4, immature-looking mast cells with a few granules started to appear, followed by a gradual increase in mast cells that reached very high levels after 8 wk, up to 40 to 200 times the normal. The pulmonary Hi and 5-HT content increased concomitantly from 6 and 1 micrograms/g to a maximum of 200 and 18 micrograms/g, respectively. These high levels of amine content and mast-cell densities greatly exceed those of any normal tissue. There was a strong correlation between the Hi and 5-HT content in both normal (r = 0.87) and irradiated (r = 0.93) lung tissue, as well as between the mast-cell density and amine content after irradiation (r = 0.86), thereby indicating that both amines derived from mast cells. The Hi/5-HT quotients were much lower in both normal and irradiated lung tissue (5 and 9, respectively) than in other tissues where these amines are stored in mast cells, or in isolated peritoneal mast cells (43). This relatively higher 5-HT content in pulmonary mast cells suggests that this amine performs a specific function in the lung. PMID- 8417752 TI - Localization of immunoreactive tissue kallikrein in human trachea. AB - Tissue kallikrein is the major kininogenase detected in bronchoalveolar lavage fluids from asthmatics and may play a particularly important role in kinin generation during asthma. The present study was undertaken to determine the source of tissue kallikrein in the human lower airways. Specific antisera to human tissue kallikrein were used to localize this enzyme by immunocytochemistry in human trachea. Immunoreactive tissue kallikrein was localized in submucosal glands of the lamina propria but was not detected in epithelial cells or goblet cells. Specific staining for tissue kallikrein was not detected in all cells of the submucosal glands but was restricted to cells forming demilunes in the distal portions of the glands. When consecutive serial sections of submucosal glands were alternately stained using antiserum to tissue kallikrein and a periodic acid Schiff stain (to detect mucus), it was revealed that immunoreactive tissue kallikrein was present only in serous cells and not in mucus cells. The localization of tissue kallikrein to the serous cells of submucosal glands should facilitate studies to regulate the release of this enzyme. Regulation of tissue kallikrein release may provide a mechanism to reduce kinin generation during asthma. PMID- 8417753 TI - Interleukin-8 expression in normal nasal epithelium and its modulation by infection with respiratory syncytial virus and cytokines tumor necrosis factor, interleukin-1, and interleukin-6. AB - Inflammation in nasal and airway tissue caused by allergens, microbial infection, and air pollution are likely to be regulated by inflammatory mediators produced by airway epithelial cells. We have therefore investigated the baseline expression of a number of cytokine genes known to be important inducers and modulators of inflammation, in freshly isolated human nasal epithelium. Cells were obtained by superficial scraping of turbinate tissue, and cDNA for polymerase chain reaction (PCR) amplification was reverse-transcribed directly from lysates of 3 x 10(3) to 5 x 10(3) epithelial cells using random hexamers. Constitutive expression of relatively high levels of interleukin-8 (IL-8) mRNA but undetectable levels (< 1 mRNA copy/cell) of granulocyte/macrophage colony stimulating factor (GM-CSF), IL-6, IL-1, or tumor necrosis factor (TNF) mRNA were found after PCR amplification of the cDNA. IL-8 protein, but not IL-6, was identified in the nasal epithelial cells by immunocytochemistry. Infection with respiratory syncytial virus (RSV) or stimulation of nasal epithelium for 4 h with TNF or IL-1 in vitro resulted in a 4- to 10-fold increase in IL-8 mRNA expression but not in the expression of detectable levels of mRNA for the other cytokines. IL-8 was secreted by RSV-, IL-1-, and TNF-stimulated as well as unstimulated nasal epithelial cells after 6 to 20 h of culture. Neither IL-6, GM-CSF, nor TNF activity/immunoreactivity was detectable in the culture supernatants. Thus, it appears that IL-8 is a major cytokine of human nasal epithelium, constitutively expressed and readily secreted upon virus infection or stimulation with IL-1 and TNF. PMID- 8417754 TI - Granulocyte/macrophage colony-stimulating factor stimulates human polymorphonuclear leukocytes to produce interleukin-8 in vitro. AB - Interleukin-8 (IL-8) is a potent chemotactic factor for polymorphonuclear leukocytes (PMN). Here we examine whether PMN synthesize and release IL-8 in response to stimulation by selected inflammatory cytokines. PMN isolated from normal heparinized peripheral human blood were incubated in RPMI culture medium at 37 degrees C in 5% CO2, with and without granulocyte/macrophage colony stimulating factor (GM-CSF). The culture supernatants were tested for chemotactic activity using a modified Boyden chamber. Immunoreactive IL-8 protein was measured by ELISA with a monoclonal antibody specific for IL-8. GM-CSF (0.01 to 50 ng/ml) stimulated PMN to produce chemotactic activity in a dose- and time dependent manner. The amount of chemotactic activity reached maximal levels after 3 h of incubation with GM-CSF. Treatment of culture media supernatants with rabbit antiserum against IL-8 blocked the GM-CSF-induced chemotactic activity. IL 8 protein concentrations detected by ELISA closely paralleled the chemotactic bioactivity in both the dose-response and kinetic studies. Northern blot analysis of total RNA from PMN using a 30 mer oligonucleotide complementary to mRNA for IL 8 yielded a single 1.6-kb band. Its intensity increased 4-fold 2 h after treatment of PMN with GM-CSF. These data suggest that peripheral blood PMN can be stimulated by GM-CSF to synthesize and secrete bioactive IL-8. Since both IL-8 and GM-CSF accumulate in sites of acute inflammation, PMN may induce IL-8 gene expression in response to GM-CSF and thereby amplify the acute inflammatory response by recruiting additional PMN into inflammatory sites. PMID- 8417755 TI - Increases in activated T lymphocytes, eosinophils, and cytokine mRNA expression for interleukin-5 and granulocyte/macrophage colony-stimulating factor in bronchial biopsies after allergen inhalation challenge in atopic asthmatics. AB - Immunohistology and in situ hybridization were used to evaluate the presence, activation status, and cytokine mRNA profile of cells in the bronchial mucosa during human allergen-induced asthma. Fifteen atopic asthmatic subjects underwent inhalation challenge with allergen and with allergen diluent, performed in random order separated by an interval of at least 3 wk. Bronchial biopsies were obtained 24 h after challenge. Immunostaining revealed increases in the numbers of secreting eosinophils (EG2+; P < 0.05) and in interleukin-2 receptor (IL-2R) positive cells (CD25+; P < 0.01) after allergen compared with diluent challenge. No differences were observed in the numbers of total leukocytes (CD45+), T lymphocytes (CD3+, CD4+, and CD8+), elastase-positive neutrophils, macrophages (CD68+), or mast cell subtypes (MCT+ or MCTC+). In situ hybridization revealed significant increases in the numbers of cells expressing mRNA for IL-5 (P < 0.02) and granulocyte/macrophage colony-stimulating factor (P < 0.01) after allergen compared with diluent challenge. A significant inverse relationship was observed between the number of cells expressing mRNA for IL-4 and for interferon-gamma (r = -0.75, P < 0.02). The results support the view that cytokines possibly from activated T lymphocytes may contribute to local eosinophil accumulation during allergen-induced asthma. PMID- 8417756 TI - Protective effect of oleoyl peptide conjugates against elastolysis by neutrophil elastase and kappa elastin-induced monocyte chemotaxis. AB - Elastin can impair the human neutrophil elastase (HNE) inhibitory capacity of elastase inhibitors. We synthesized oleoyl-alanyl-alanyl-prolyl-valine (Ol-Ala Ala-Pro-Val-OH) (oleoyl peptide) and the amides (NH2 and NH-C3H7) of this peptide and studied their HNE-inhibitory potencies using succinyl-alanyl-alanyl-alanine-p nitroanilide (Suc-Ala-Ala-Ala-pNA) or 3H-labeled elastin as substrates, as well as cryostat sections of rabbit skin as an ex vivo substrate. Using Suc-Ala-Ala Ala-pNA, Ol-Ala-Ala-Pro-Val-OH had an IC50 of 3 microM. When the COOH terminal of the oleoyl peptide was derivatized to amide forms, the compound lost its ability to interact with HNE while keeping its elastin-protecting function: IC50 values for NH2 and NH-C3H7 derivatives were 22 and 17 microM, respectively. Also, the HNE-inhibitory capacity of Ol-Ala-Ala-Pro-Val-OH was only reduced 2-fold by using elastin as a substrate. This decrease was much lower than those determined with other HNE inhibitors of similar potency and could be accounted for by the ability of oleoyl peptide to bind to elastin. Cryostat sections of rabbit skin were also used as an ex vivo substrate for assessing the elastin-protecting property of Ol Ala-Ala-Pro-Val-OH. Preincubating HNE and oleoyl peptide before application to tissue sections led to an IC50 of 8 microM, close to the value determined with elastin as a substrate. Treatment of sections with oleoyl peptide before adding HNE gave a lower IC50 (4 microM).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417757 TI - Resistance of human tracheal epithelial cells to killing by neutrophils, neutrophil elastase, and Pseudomonas elastase. AB - The airway disease of cystic fibrosis (CF) is characterized by massive polymorphonuclear leukocyte (PMN) infiltration and the presence of variable but substantial quantities of uninhibited elastases derived from both PMNs and the common infecting organism Pseudomonas aeruginosa. In order to determine whether these agents inflict fatal injury on the airway epithelium, we exposed primary cultures of human tracheal epithelial (HTE) cells to activated PMNs, PMN elastase (PMNE), and elastase from P. aeruginosa (PSE) and monitored cytotoxicity by 51Cr release assay. Activated PMNs did not kill HTE cells, and fewer than 2% of the added PMNs adhered to the HTE cell layer. Pretreatment of HTE cells with lipopolysaccharide, incubation of PMNs with cytochalasin B, or increasing the incubation period to 8 h did not increase PMN adherence or target cell killing. However, poor PMN adherence was not by itself responsible for lack of cytotoxicity, since PMNs were not cytotoxic for 9HTEo- cells, a HTE cell line to which PMNs adhere in large numbers. Purified PMNE, but not exogenous H2O2, caused a small but significant increase in cytotoxicity after 6 h of incubation, but only at the highest concentrations tested (10 and 50 micrograms/ml). The PMNE remained fully active throughout the incubation period. Some detachment of the cell layer occurred after 4 h of incubation with 10 micrograms/ml PMNE. PSE at concentrations > 1 micrograms/ml also caused slight cytotoxicity and removal of the cell layer from the culture substratum. Ultrastructural studies showed only minor cytoplasmic vacuole formation. We conclude that cultured HTE cells are resistant to cytolysis by PMNs and elastases. PMID- 8417758 TI - Migration of bovine bronchial epithelial cells to extracellular matrix components. AB - Migration of epithelial cells is an important feature of wound healing. Components of extracellular matrix stimulate migration of other cells, and we hypothesized that basement membrane components, laminin and type IV collagen, stimulate migration of bovine bronchial epithelial cells (BBEC). BBEC cultured for 3 days were used in migration assays using the blindwell chamber technique. BBEC migrated in a concentration-dependent manner to laminin, type IV collagen, and fibronectin. "Checkerboard" analysis demonstrated that the migration was directional for each of the components studied. We also evaluated the stimulatory activity of fibronectin fragments. A fragment containing the RGDS peptide was stimulatory of migration while other fragments were much less so, suggesting a role for RGDS-sensitive, integrin-mediated mechanisms. In order to evaluate whether bound or soluble matrix components were required to direct cell migration, we performed haptotaxis assays with precoated filters in the migration chambers. Fibronectin-precoated filters demonstrated significant stimulation of migration, suggesting that some of the migration in our original assays with fibronectin present in the lower chambers could be termed haptotactic. laminin- and type IV collagen-precoated filters were less active. When insulin was used as a chemoattractant, the fibronectin-precoated filters were more facilitatory of migration than the other matrix coatings. In summary, BBEC can chemotactically migrate to extracellular matrix components. Thus, the composition of the provisional matrix that forms at sites of epithelial injury may play an important role in the repair processes that occur after injury to the bronchial epithelium. PMID- 8417759 TI - Human fetal lung fibroblasts promote invasion of extracellular matrix by normal human tracheobronchial epithelial cells in vitro: a model of early airway gland development. AB - Epithelial invasion of extracellular matrix (ECM) is important during lung development and the pathogenesis of bronchogenic neoplasms. Airway submucosal gland development begins when clusters of surface epithelial cells invade the lamina propria between the tenth and thirtieth weeks of fetal life. The factors regulating this transient normal invasive behavior are unknown. We observed that normal human tracheobronchial epithelial (HTBE) cells from adult necropsy specimens penetrate collagen matrices when co-cultured with human fetal lung fibroblasts (HFLF). Invading clusters of epithelial cells resembled primordial glands, forming tubular structures and undergoing dichotomous branching. Using 48 different tracheobronchial specimens, we compared paired cultures of HTBE cells without (control) and with HFLF co-culture. Fixed, vertically sectioned culture substrates were examined, and invaginated epithelial cell clusters as well as total invading HTBE cells were counted. The co-cultured condition resulted in significantly more epithelial invaginations per two sections (4 +/- 1 versus 22 +/- 3, P < 0.0005) and more invading HTBE cells per two sections (31 +/- 9 versus 194 +/- 27, P < 0.0005) than controls. Epithelial invasion was noted by 36 h in culture and was greatest at HTBE cell/HFLF ratios near 1 and HFLF passages between 10 and 16. Epithelial cells co-cultured with a fibroblast cell line derived from the adult bronchiole showed no increase in ECM invasion compared with controls. These results demonstrate that fetal mesenchymal cells are capable of promoting invasive behavior in mature epithelial cells in vitro. Given the fibroblast type and passage specificity, this model should prove useful for investigating the cellular and molecular regulation of epithelial ECM invasion. PMID- 8417760 TI - Mast cells and fibrosis--who's on first? PMID- 8417761 TI - In vitro effects of pentoxifylline on smooth muscle cell migration and blood monocyte production of chemotactic activity for smooth muscle cells: potential therapeutic benefit in the adult respiratory distress syndrome. AB - The adult respiratory distress syndrome (ARDS) is a severe lung condition characterized by an acute lung injury leading to a massive intra-alveolar fibrosis with rapid lung failure. ARDS intra-alveolar fibrosis results from the migration of mesenchymal cells (mainly smooth muscle cells [SMC]) into the alveoli through alveolar epithelial basement membrane gaps resulting from the injury. SMC migration is followed by their replication and production of extracellular matrix, which leads to fibrosis. Thus, any pharmacologic agent able to prevent SMC migration should prevent, at least in part, intra-alveolar fibrosis. SMC migration is thought to be due to the presence, in the alveolar spaces, of chemotactic factors for mesenchymal cells, such as fibronectin and platelet-derived growth factor (PDGF). The local presence of these chemotactic factors can be due to plasmatic leakage, platelet degranulation, and mononuclear phagocyte activation. Pentoxifylline is a methylxanthine interacting with the biology of several types of cells, including red blood cells, neutrophils, blood monocytes, and endothelial cells. Pentoxifylline prescription has been suggested in ARDS with respect to its activity on neutrophils, its inhibition of tumor necrosis factor-alpha (TNF) release by mononuclear phagocytes, and its prevention of TNF-induced lung injury. Since pentoxifylline can modulate the migration of several cell types, we hypothesized that it could interfere with mesenchymal cell migration. SMC migratory response was measured in vitro with modified Boyden chemotactic chambers in the presence of PDGF, fibronectin, "platelet extract," and activated blood monocyte supernatants. Pentoxifylline, at therapeutic levels, significantly reduced SMC migration in response to the presence of these chemotactic activities.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417763 TI - New beginnings. PMID- 8417762 TI - Characterization and immunohistochemical localization of the 15 kD protein isolated from rat lung lamellar bodies. AB - We have characterized a protein of approximately 15 kD (lb15) derived from rat lung lamellar bodies, and then sequenced the first 42 residues. Following the normal isopycnic sucrose gradient ultracentrifugation, we diluted the band containing the crude lamellar body fraction with an equal volume of cold distilled water and further centrifuged it at 2,000 x g for 30 min to pellet a fraction of lamellar bodies. Under the electron microscope, this fraction appeared intact and highly purified. When this fraction was subjected to polyacrylamide gel electrophoresis, the major protein was one of 15 kD, regardless of whether the fraction was extracted or unextracted, reduced or unreduced; only a small amount of 35 kD protein was detected with Coomassie Blue staining. Disruption of lamellar bodies revealed that the limiting membrane was particularly enriched with lb15. Immunohistochemistry indicated that lb15 was present in lamellar bodies and tubular myelin, suggesting it was secreted along with the lipid. Amino acid analysis revealed a protein with 13.5% basic and 10.6% acidic residues. The N-terminal appeared particularly highly charged, with 32% of the charged residues in the first 14 amino acids. The lb15 protein is identical to rat lysozyme for the first 23 residues, with the important exception of residue 6, which is histidine in lb15 and cysteine in lysozyme. Residue 24 was not identified. Lb15 was also present in lavage material. We conclude that lb15 is the major protein in rat lung lamellar bodies, has a highly charged N terminal, and shares some sequence homology with rat lysozyme.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417764 TI - Dentistry--the challenge of the nineties. AB - Members of the dental profession over the past decade have been forced to recognise their identity as small businesses as well as healthcare professionals. This recognition has been painful for many. PMID- 8417765 TI - 'Safety in radiation'. PMID- 8417766 TI - 'Trends in the incidence of oral carcinoma in Northern Ireland 1975-89'. PMID- 8417767 TI - Caries and periapical pathology in an unerupted tooth. PMID- 8417768 TI - 'Teamwork of a different kind'. PMID- 8417769 TI - Hidden sugars. PMID- 8417770 TI - Toothpaste for dogs. PMID- 8417771 TI - Dental anomalies--are adhesive castings the solution? AB - Adhesive cast restorations placed without tooth preparation were used to restore the teeth of patients affected by either amelogenesis or dentinogenesis imperfecta. Twelve patients aged between 7 and 18 years were treated and in total 64 cast restorations were placed. At the time of cementation, the vertical dimension was increased by the thickness of restorative material placed on the functioning surfaces of the restored teeth. Full occlusion was restored for all individuals, with only one casting being lost. The castings have been placed for intervals between 10 and 54 months. PMID- 8417772 TI - Parental presence during treatment of the child patient: a study with British parents. AB - A clinical study was carried out to determine if parental presence during dental treatment alters the child's behaviour when compared with parental absence. Thirty-two children who attended with a parent for a dental recall visit were randomly assigned to a parent present or a parent absent group. The age of the children ranged from 4 to 12 years. Twenty-three mothers and nine fathers attended during the study. Those parents absent during the visit viewed proceedings from behind a one-way mirror. Each visit was standardised in procedure and recorded on video tape. The results showed that 4 to 8-year-old children exhibited significantly more negative behaviour, regardless of parental presence, than the 9 to 12-year-olds. The presence of the parent did not lead to significantly greater negative behaviour. The parent's need to see what happened during the visit and the child's need to have a parent present were the most important reasons for having a parent present. Parents viewing from behind a one way mirror were as satisfied with their position as parents who had been actually present in the surgery. PMID- 8417773 TI - The control of cross-infection in UK clinical dentistry in the 1990s: immunisation against hepatitis B. AB - Thirty thousand dentists and clinical ancillary staff in the UK were surveyed in July 1991 to assess the current state of immunisation against hepatitis B virus (HBV). About 11,000 responded immediately and nearly 94% of these had been, or were being, immunised against HBV. Nearly all dentists and therapists (94% in each group), 95% of hygienists, and 96% of dental surgery assistants had been immunised. However, one half of the respondents were at or approaching 5 years post-immunisation, the time when booster immunisation is recommended. About 53% of most responding clinical dental staff had been vaccinated 4 to 5 years before the survey, but, of these, nearly 81% had not had booster immunisation. Booster immunisation will now be indicated for most dental clinical personnel. PMID- 8417774 TI - The practice meeting. AB - In any really successful business there is one common factor which stands out- effective communication. When management, staff and clients communicate effectively a business, small or large, has laid the foundation stone for success. PMID- 8417775 TI - Of rings and clearer information. PMID- 8417776 TI - Mike Grace talks to the President of the General Dental Council, Sir David Mason. AB - Dentistry, as with all things, moves in the direction of the thinking of those people within it, whether they are members of the dental team working within their surgeries, various colleagues in practice and on the many dental committees, teachers involved with undergraduates, people in the dental industry and trade, technicians in laboratories or leaders of the many organisations. This column will explore the thoughts and attitudes of many of these people--both those in the public limelight and those as yet unknown. To celebrate the launch of 'Interview with the Editor' who better to feature than the President of the General Dental Council, Sir David Mason. In this interview he shares his own thoughts and hopes for the future, thoughts that will influence the lives of most of the profession. PMID- 8417777 TI - The binding change mechanism for ATP synthase--some probabilities and possibilities. PMID- 8417778 TI - AHA- heterodimer of a class-2 uncoupler: pentachlorophenol. AB - AHA- heterodimers formed by association of neutral molecules of weak acid (HA) with its conjugate anion (A-) have been proposed to be the charged membrane permeable species of class-2 uncouplers. Past attempts to extract and identify AHA- heterodimers failed. We have measured optical spectra of HA+A- (1:1) solutions of pentachlorophenol (PCP) in various solvents and in the presence of PC liposomes. Optical studies were supplemented by nuclear magnetic resonance measurements of HA+A- (1:1) solutions of PCP in dichloroethane to gain insight into the formation of AHA- species in lipid membranes. From these experiments, we found evidence for AHA- formation in non-hydrogen-bonding solvents, then reported the AHA- formation constant Kf and the molar absorptivity epsilon AHA-(lambda). Kf decreases with increasing dielectric constant, kappa, from 1210 +/- 130 M-1 for dichloroethane (kappa 10.7), to 340 +/- 34 M-1 for acetonitrile (kappa 37.5); Kf also decreases with increasing concentration of water. In hydrogen-bonding solvents, octanol (kappa 10.3) and methanol (kappa 33.5) and in liposomes, AHA- heterodimers are not observed optically. We estimate Kf for PCP in lipid bilayers from a combination of data on membrane electrical conductivity and surface density of adsorbed PCP. Our estimate for lipid bilayer, 0.005 < Kf < 0.5 M-1, is consistent with our inability to detect the AHA- species optically in liposome suspensions. We propose that penetration of water into the membrane inhibits formation of AHA- in lipid bilayers. PMID- 8417779 TI - Fumarate reductase activity of bovine heart succinate-ubiquinone reductase. New assay system and overall properties of the reaction. AB - A simple system for aerobic assay of the quinol-fumarate reductase reaction catalyzed by purified soluble bovine heart succinate-ubiquinone reductase in the presence of NADH, NAD(P)H-quinone reductase (DT-diaphorase) and an appropriate quinone is described. The reaction is inhibited by carboxin, suggesting that the same quinone/quinol binding site is involved in electron transfer from succinate to ubiquinone and from ubiquinol to fumarate. The kinetic properties of the reaction in both directions and comparative affinities of the substrate binding sites of the enzyme to substrates (products) and competitive inhibitors are reported. Considerable difference in affinity of the substrates binding site to oxaloacetate was demonstrated when the enzyme was assayed in the direct and reverse directions. These results were taken to indicate that the oxidized dicarboxylate-free enzyme is an intermediate during the steady-state succinate ubiquinone reductase reaction, whereas the reduced dicarboxylate-free enzyme is an intermediate of the steady-state ubiquinol-fumarate reductase reaction. No difference in the reactivity of the substrate-protected cysteine and arginine residues was found when the pseudo-first-order rate constants for N ethylmaleimide and phenylglyoxal inhibition were determined for oxidized and quinol-reduced enzyme. Quinol-fumarate reductase activity was reconstituted from the soluble succinate dehydrogenase and low-molecular-mass ubiquinone reactivity conferring protein(s). No reduction of cytochrome b was observed in the presence of quinol generating system, whereas S-3 low temperature EPR-detectable iron sulfur center was completely reduced by quinol under equilibrium (without fumarate) or steady-state (in the presence of fumarate). No significant reduction of ferredoxin type iron-sulfur centers was detected during the steady-state quinol-fumarate oxidoreductase reaction. The data obtained eliminate participation of cytochrome b in the quinol-fumarate reductase reaction and show that the rate limiting step of the overall reaction lies between iron-sulfur center S-3 and lower midpoint potential redox components of the enzyme. PMID- 8417780 TI - Ca(2+)-induced, phospholipase-independent injury during reoxygenation of anoxic mitochondria. AB - Reoxygenation of rat-liver mitochondria after anoxic incubation induced release of matrix proteins. As assessed by release of a matrix enzyme, it was proportional to the rate of H2O2 production. The release was not observed with low concentrations of extramitochondrial free Ca2+, indicating a Ca(2+)-dependent pathway. Phospholipase A2 was not involved in the reoxygenation injury, because non-esterified fatty acids did not increase on reoxygenation even when re acylation was inhibited and because inhibitors of phospholipase A2 had little effect on enzyme release. Cyclosporin A, ATP, ADP and inhibitors of pyridine nucleotide oxidation had a protective effect, strongly suggesting involvement of so-called Ca(2+)-dependent permeability transition. Ca2+ was also released from reoxygenated mitochondria and inhibition of reuptake of released Ca2+ attenuated the enzyme release. Similar releases of aspartate aminotransferase and Ca2+ were observed with mitochondria in an oxygen radical-generating system, hypoxanthine and xanthine oxidase. In this system, lecithin-cardiolipin liposomes also released entrapped Ca2+ without disruption of the membrane. From these results, we conclude that during reoxygenation, Ca2+ release and subsequent reuptake induced permeability transition of mitochondria, resulting in reoxygenation injury. PMID- 8417781 TI - Influence of the mitochondrial outer membrane and the binding of creatine kinase to the mitochondrial inner membrane on the compartmentation of adenine nucleotides in the intermembrane space of rat heart mitochondria. AB - The influence of the mitochondrial outer membrane and that of the binding of creatine kinase to the mitochondrial inner membrane on the compartmentation of adenine nucleotides in the intermembrane space of rat heart mitochondria were investigated under conditions of maximal rates of mitochondrial creatine kinase. To this end, experiments were performed in reconstituted systems consisting of functionally intact rat heart mitochondria or mitoplasts and pyruvate kinase, both competing at ADP formed by mitochondrial creatine kinase in the presence of high concentrations of adenine nucleotides. Results showed that removal of the mitochondrial outer membrane diminished the compartmentation effects, supporting the relevance of the mitochondrial outer membrane to compartmentation effects in the intermembrane space. Compartmentation effects were clearly seen even in KCl containing media which release the creatine kinase from the inner membrane, indicating that the localization of the ADP-regenerating enzyme in the mitochondrial intermembrane space within the outer membrane is a prerequisite for dynamic compartmentation of adenine nucleotides. Under these conditions, the binding of creatine kinase to the inner membrane is of minor importance. Lowering the ATP concentrations diminished the extent of AdN compartmentation due to decreased creatine kinase rates. Under these conditions of low AdN fluxes, the release of creatine kinase from the inner membrane entailed another decrease in the compartmentation. PMID- 8417782 TI - Mechanisms of thymus homing. PMID- 8417783 TI - Analysis of the expression of the hybrid gene bcl-2/IgH in follicular lymphomas. AB - To investigate the clinical and biologic significance of the circulating t(14;18) carrying cells in follicular lymphoma (FL) patients, we analyzed the mbr/JH junction of the hybrid bcl-2/IgH gene simultaneously at the DNA and RNA levels by polymerase chain reaction (PCR) in 37 peripheral blood samples from 37 patients in different remission status: 4 before treatment, 8 during treatment, and 25 in complete remission (CR). Of these 37 patients, 22 were positive either at the DNA or RNA level (8 with active disease and 14 in CR). Among these positive patients, RNA was more often negative for patients in CR (9 of 14 [64%]) than for patients with active disease (2 of 8 [25%]; Fisher's exact test, P = .09). Among the 14 patients in CR with residual disease, 2 of 5 with RNA positivity relapsed, whereas 1 of 9 with RNA negativity and DNA positivity relapsed with a median follow-up after sample collection of 8 months (range, 4 to 18 months). Simultaneous analysis of the bcl-2/IgH gene at the DNA and RNA level showed heterogeneous patterns of PCR positivity in regards to the evaluation of the biologic activity of the t(14;18)-carrying cells. A larger study and long-term follow-up will help in determining whether the expression patterns in turn reflect the functional status of disease activity in FL patients. PMID- 8417784 TI - p53 gene mutations in multiple myeloma are associated with advanced forms of malignancy. AB - The frequency and type of p53 gene mutations was investigated in a series of 52 cases of multiple myeloma (MM) representative of the different clinical phases and forms of the disease (indolent, 12 cases; chronic, 24 cases; acute/leukemic, 16 cases). DNAs were analyzed for p53 gene mutations in exons 5 to 9 by polymerase chain reaction (PCR), single-strand conformation polymorphism (SSCP), and direct sequencing of PCR-amplified fragments. Point mutations were detected in 7 of 52 patients (13%) (5 at exon 8; 1 at exon 6; 1 at exon 7), and were specifically associated with the more advanced and clinically aggressive acute/leukemic forms of MM (7 of 16 [43%].) Three of the mutated cases had been evaluated at clinical presentation in earlier phases of the disease (indolent or chronic) and were found to be negative for p53 mutation. Moreover, three patients with p53 mutation had not received chemotherapy at the time of investigation. These results support the notion that the development of MM is a multistep process and suggest that alterations in the p53 gene may represent an important late event in MM tumor progression. PMID- 8417785 TI - FVT-1, a novel human transcription unit affected by variant translocation t(2;18)(p11;q21) of follicular lymphoma. AB - Variant t(2;18) and t(18;22) chromosome translocations observed in B-cell chronic lymphocytic leukemias and in follicular lymphomas have been reported to consistently involve the 5' region of the BCL-2 gene on chromosome 18 and various regions on the lg light chain loci. We show here that a variant t(2;18)(p11;q21) translocation observed in a case of follicular lymphoma leads to the juxtaposition of a J kappa segment to a chromosome 18 transcriptional unit located 10 kpb upstream of the BCL-2 locus. The cDNA of this new evolutionarily conserved gene, termed FVT-1 for follicular-variant-translocation gene, codes for a putatively secreted protein of 36 Kd that is not homologous with any described protein. The FVT-1 gene is weakly expressed in all the analyzed normal hematopoietic tissues but a very high rate of transcription is observed in some T cell malignancies and in phytohemagglutinin-stimulated lymphocytes. The proximity of FVT-1 to the BCL-2 locus suggests that in the t(14;18) currently observed in follicular lymphomas, both genes would participate in the tumoral process. PMID- 8417786 TI - Bcl-2 oncoprotein blocks chemotherapy-induced apoptosis in a human leukemia cell line. AB - Previous studies have shown that the bcl-2 gene encodes a mitochondrial protein that contributes to neoplastic cell expansion primarily by promoting cell survival through interference with "programmed cell death" (PCD), also termed "apoptosis." Because many chemotherapeutic drugs are capable of initiating pathways leading to apoptosis, we determined whether deregulated bcl-2 expression could render cells resistant to several drugs commonly used in the treatment of non-Hodgkin's lymphomas, including dexamethasone (DEX), methotrexate (MTX), 1 beta-D-arabinofuranosyl-cytosine (Ara-C), etoposide (VP-16), vincristine (VC), cisplatin (CP), and hydroperoxycyclophosphamide (4-HC). For these experiments, we achieved high levels of p26-Bcl-2 protein production in a human pre-B-cell leukemia line 697 by stable infection with a recombinant bcl-2-containing retrovirus and then compared these cells with control virus-infected 697 cells. Control 697 cells were induced to undergo apoptosis by all drugs tested as defined by DNA degradation into oligonucleosomal-length fragments, cell shrinkage, and subsequent cell death. In contrast, 697 cells with elevated Bcl-2 protein levels exhibited strikingly prolonged cell survival and markedly reduced DNA fragmentation when cultured in the presence of these antineoplastic agents. Although high levels of Bcl-2 protein protected 697 cells from the acute cytotoxic effects of DEX and the other drugs tested, Bcl-2 did not prevent these drugs from suppressing the proliferation of 697 cells. However, when 697 cells were treated with DEX or MTX for 3 days, then washed and cultured in semisolid media without drugs, bcl-2-virus-infected cells gave rise to colonies at much higher frequencies than 697 cells stably infected with control virus. These results indicate that by protecting 697 leukemic cells from the acute cytotoxicity of DEX and some other chemotherapeutic drugs, high levels of p26-Bcl 2 can create the opportunity for re-initiation of cell growth when drugs are withdrawn. The findings may be relevant to clinical correlative studies of non Hodgkin's lymphoma patients that have found an association between worse prognosis and bcl-2 gene rearrangements or t[14;18] translocations. PMID- 8417787 TI - The ABL-BCR fusion gene is expressed in chronic myeloid leukemia. AB - Although the BCR-ABL hybrid gene on chromosome 22q-plays a pivotal role in the pathogenesis of chronic myeloid leukemia (CML), little is known of the reciprocal chimeric gene, ABL-BCR, formed on chromosome 9q+. By reverse transcription/polymerase chain reaction amplification (RT/PCR) we have detected ABL-BCR mRNA in cells from 31 of 44 BCR-ABL positive CML patients and 3 of 5 CML cell lines. Of the 34 positive samples, 31 had classical t(9;22) (q34;q11) translocations; in 3 samples there was no Philadelphia (Ph) and/or 9q+ chromosomes. ABL-BCR expression consisted of ABL(Ib)-BCR mRNA in 26 patients and of both ABL(Ib)-BCR and ABL(Ia)-BCR mRNA species in 6 patients. The ABL-BCR transcripts encoded one or, more rarely, both of the two potential junctions, designated ABL-b3 and ABL-b4, which differed in size by 75 bp. In 2 patients, the BCR exon b3 was not present in either the BCR-ABL or the corresponding ABL-BCR transcript, whereas in 5 patients exon b3 was present in both transcripts. Direct sequencing of PCR fragments representing the full-length coding sequence of ABL BCR cDNAs type Ib-b3, Ia-b3, Ib-b4, and Ia-b4 showed an open reading frame predicted to encode fusion proteins of 370 to 414 amino-acids. If an ABL-BCR gene product is produced in CML cells, it may be relevant as a mechanism for deregulating the GTPase activating protein (GAP) function of BCR. PMID- 8417788 TI - Release of elastase from monocytes adherent to a fibronectin-gelatin surface. AB - Fibronectin (Fn) is a circulating and extracellular matrix glycoprotein that may serve to facilitate phagocytosis because of its ability to bind many inflammatory ligands and to a monocyte receptor. Fn fragments have been shown in many systems to have augmented properties over those of native Fn. We show in this report that although Fn fragments did not cause elastase release from monocytes in suspension, fragments did cause elastase release from monocytes that were first bound to Fn-gelatin surfaces. An amino-terminal 29-Kd and a 140-Kd integrin binding fragment were half-maximally active at 100 nmol/L, whereas the Arg-Gly Asp-Ser integrin-recognition peptide was half-maximally active at 100 mumol/L. Fluid-phase Fn was ineffective yet blocked the activity of the Fn fragments. Complexing of Fn with gelatin or with heparin partially removed the blocking effect of Fn. Similar results were obtained with U-937 cells. Substitution of the Fn-gelatin surface with bovine articular cartilage also promoted elastase release. Therefore, in conditions in vivo in which monocytes bind to tissue surface, a high ratio of Fn fragments to native Fn may upregulate certain monocyte activities such as protease release. PMID- 8417789 TI - Glucose-6-phosphate isomerase deficiency associated with nonspherocytic hemolytic anemia in the mouse: an animal model for the human disease. AB - The first two mutations causing hereditary glucose-6-phosphate isomerase (GPI) deficiency associated with chronic nonspherocytic hemolytic anemia in nonhuman mammals are described in the mouse. As in humans, the hemolytic syndrome, which is characterized by a diminished erythrocyte number, lower hematocrit, lower hemoglobin, higher number of reticulocytes and plasma bilirubin concentration, as well as increased liver- and spleen-somatic indices, was exclusively manifested in homozygous mutants. In comparison with wild type, heterozygous individuals exhibited neither hematologic differences nor alterations of other physiologic parameters, including plasma concentration of glucose, pyruvate and lactate, body weight, organo-somatic indices of liver, lung, kidney, spleen, and heart, as well as viability. Glycolytic intermediates, adenine nucleotides, and metabolic rate were not significantly altered in erythrocytes from heterozygotes. On the contrary, if allowance is made for the young erythrocyte population, homozygous mutant erythrocytes showed an increased concentration of glucose-6-phosphate and normal or decreased concentrations of glycolytic metabolites following the enzymatic block. The concentration of adenosine triphosphate and the glycolytic rate also appeared to be reduced. Homozygous anemic mice showed hepatosplenomegaly and typical adaptations to hypoxia, such as an elevated heart somatic index and, for one mutant line, an enhanced lung-somatic index. Further, these animals were characterized by a marked reduction of body weight and an increase of lethality both correlated with the degree of enzyme deficiency in tissues. The latter findings were attributed to a reduced glycolytic capability of the whole organism caused by the enzyme defect in tissues, rather than representing secondary consequences of GPI deficiency in erythrocytes. The similarity in physicochemical and kinetic properties of the mutant murine proteins reported earlier with those of allozymes found in human GPI deficiency, as well as the comparable metabolic and physiologic consequences of this enzyme defect in mice and humans support that these murine mutants are excellent animal models for the human disease. PMID- 8417790 TI - Mode of action of iron (III) chelators as antimalarials: I. Membrane permeation properties and cytotoxic activity. AB - We have designed two subfamilies of lipophilic iron (III) chelators previously termed reversed siderophores (RSFs). The agents display physicochemical properties that favor extraction of iron beyond membrane barriers of Plasmodium falciparum-infected red blood cells. We studied the in vitro antimalarial potency of RSFs and their relationship to the membrane permeation properties of these agents. The mode of RSF action involves: (1) fast access to intracellular compartments of parasitized cells; (2) selective and high-affinity chelation of iron (III) from parasitized cells; (3) fast exit from cells after iron (III) complexation; and (4) exertion of cell damage on parasites exposed for 3 to 5 hours to drugs, irrespective of the stage of parasite development. These results suggest that on reaching a critical intraerythrocyte target, RSFs induce an iron deficit that parasites in general, and rings in particular, have limited capacity to restore. PMID- 8417791 TI - Transgenic mice expressing human sickle hemoglobin are partially resistant to rodent malaria. AB - The polymorphic frequency of the gene for beta s-globin involved in the generation of sickle trait and sickle cell anemia in the human population is caused by the enhanced resistance of sickle trait individuals to Plasmodium falciparum malaria, as supported by epidemiologic and in vitro studies. However, the mechanism for the protective effect of sickle hemoglobin in vivo has not been fully defined. The generation of transgenic mice expressing high levels of human beta s- and alpha-chains has allowed us to study this phenomenon in vivo in an experimental model. We infected the transgenic beta s mice with two species of rodent malaria and found a diminished and delayed increase in parasitemia as compared with controls. This is in contrast to our previous studies involving the introduction of a beta A transgene, which does not alter the infection. The use of this model allowed us to address the question of the mechanism of protection against malaria in mice expressing sickle hemoglobin. We find that splenectomy of transgenic mice completely reverses the protection against Plasmodium chabaudi adami infection. The results reported have shown a relationship between the presence of the beta s gene product and partial resistance to malaria in an experimental model in vivo and shows that the spleen plays an important role in this protection. PMID- 8417792 TI - Characterization and quantitation of the circulating forms of serum transferrin receptor using domain-specific antibodies. AB - To characterize the nature of the immunoreactive transferrin receptor in human serum, antisera were developed to peptide sequences of the extracellular domain of human transferrin receptor between amino acids 107 and 120 and the intracellular domain between amino acids 40 and 54. Antisera against the extracellular domain exhibited reactivity against both purified intact receptor and immunopurified circulating receptor, whereas antisera against the intracellular domain reacted only with intact receptor. Using competitive binding enzyme-linked immunosorbent assays, transferrin receptor in ultracentrifuged sera from normal subjects and patients with sickle cell anemia could be detected with antisera against the extracellular but not the intracellular domain. When the pellet obtained by ultracentrifugation of these sera was assayed after solubilization in 1% teric (polyoxyethylene-9-lauryl ether), only 0.6% of total serum receptor was detected in normal subjects and 3.8% in subjects with sickle cell disease. Roughly equal amounts of this pelleted immunoactivity were detected with antibodies against the extracellular and intracellular domains. These results indicate that less than 1% of transferrin receptor in normal human sera is intact receptor consistent with an exosomal origin and that virtually all circulating transferrin receptor is in the form of a truncated extracellular domain. PMID- 8417793 TI - Rapid and simultaneous typing of hemoglobin S, hemoglobin C, and seven Mediterranean beta-thalassemia mutations by covalent reverse dot-blot analysis: application to prenatal diagnosis in Sicily. AB - The molecular lesions causing beta-thalassemia in Sicily can be subdivided into two groups. One that occurs at a 71% frequency and consists of the beta 39, IVS 1,110 and IVS 1,6 mutations and the other group at a 20% frequency comprising the -87, beta s, IVS 1,1 and IVS 2,745 mutations. The identification of all these mutations by polymerase chain reaction (PCR) and conventional dot-blot hybridization has been time consuming and expensive. In this article, we describe the implementation of the reverse dot-blot (RDB) hybridization as a rapid nonradioactive method for the identification of the nine most frequent molecular lesions in the beta-globin gene (-87, beta s, beta c, IVS 1,1, IVS 1,6, IVS 1,110, beta 39, IVS 2,1, IVS 2,745) in Sicily. Sixty prenatal diagnoses were performed by this RDB assay, each of which was confirmed by dot-blot/ASO hybridization; thus demonstrating the accuracy of the RDB. The main advantage of this assay is the rapid typing of an individual's DNA for many mutations in a single working day. Because the mutations in this assay are representative for the Mediterranean region, this mutational panel can also be extended to the screening of beta-thalassemia from other Mediterranean regions. PMID- 8417794 TI - Characterization of mixed chimerism in patients with chronic myeloid leukemia transplanted with T-cell-depleted bone marrow: involvement of different hematologic lineages before and after relapse. AB - We have characterized mixed chimerism (MC) in five patients with chronic myeloid leukemia (CML) who received transplants with T-cell-depleted bone marrow (BM) and who relapsed within 4 years after transplantation. To study the possible relation of MC with relapse, we purified different populations of leukocytes and analyzed their donor/recipient origin by a method based on polymerase chain reaction amplification of minisatellite DNA regions. Our results show that before relapse, all hematopoietic recipient cells are T cells, whereas monocytes, B, and natural killer (NK) cells are of donor origin. This observation does not appear to be specific for CML as similar results were found in two control patients with acute myeloid leukemia (AML). At the time of (CML) relapse, recipient granulocytes, monocytes, and erythrocytes appeared and progressively replaced the respective lineages of donor origin. No other lineages seemed to be involved as B cells and NK cells remained of donor origin and no significant changes in the number of recipient T cells were detected. In this respect relapse of CML after BM transplantation (BMT) seems not to be very different from the primary disease in chronic phase before transplantation. Furthermore, we conclude that after BMT, an association between mixed chimerism before relapse and the (CML) relapse does exist because both phenomena are consequences of T-cell depletion of the BM graft. However, this correlation might well be indirect as the MC caused by the recipient T cells appears to be independent of the one caused by the recurrent disease. PMID- 8417795 TI - Marrow transplantation from unrelated donors for treatment of hematologic malignancies: effect of mismatching for one HLA locus. AB - One hundred twelve patients less than 36 years old received marrow grafts from unrelated donors as treatment for hematologic malignancy. Seventy donor/recipient pairs were phenotypically identical for HLA-A, -B, and -D, while 42 had a "minor" disparity at one HLA locus. There was an increase in the risk of acute graft versus-host disease (GVHD) in patients receiving HLA-partially matched grafts compared with those receiving HLA-matched grafts (51% v 36% probability of grades III-IV acute GVHD). However, in this cohort of patients, there was no significant difference in survival (at 1.5 years, 46% v 51% for good-risk patients, 44% v 30% for poor-risk patients). This finding suggests that some degree of HLA disparity can be tolerated in young patients transplanted from unrelated donors for malignant disease, thus making transplantation an option available to larger numbers of patients. PMID- 8417796 TI - Transplantation for acute leukemia in small versus large centers. PMID- 8417797 TI - t(2;3)(p12;q27) in Hodgkin's disease of a human immunodeficiency virus-positive patient with hemophilia. PMID- 8417798 TI - Effects of recombinant human interleukin-11 on hematopoietic reconstitution in transplant mice: acceleration of recovery of peripheral blood neutrophils and platelets. AB - We have examined the effects of recombinant human interleukin-11 (rhIL-11) on the recovery of peripheral blood cell counts and proliferation of progenitors and hematopoietic stem cells (day 12 colony-forming units-spleen-CFU-S12) in vivo using a mouse bone marrow (BM) and spleen cell transplantation model. Recovery of leukocytes was accelerated in animals receiving daily administration of rhIL-11 (100 micrograms/kg/d) and reached normal levels by day 14 posttransplantation. This increased total leukocyte count reflected mainly an increase in neutrophils. Neutropenia (absolute neutrophil count [ANC] < 1,500) was present in control transplant mice for 14 to 15 days, while in the rhIL-11-treated group, neutrophils recovered to normal by days 8 to 10 and continued to increase until day 19. Animals treated with rhIL-11 had only 1 day with ANC demonstrated < 500. Correspondingly, rhIL-11 treatment increased granulocyte-macrophage progenitors (CFU-GM) derived from both spleen and BM cells. Higher doses of IL-11 increased CFU-GM nearly threefold and CFU-Mix fourfold to fivefold, while increasing burst forming units-erythroid to a lesser degree. BM and spleen cellularity were both increased in IL-11-treated mice, but no increase in CFU-S12 was noted. In addition, in vivo daily administration of IL-11 increased peripheral platelet counts by threefold over control transplant mice at day 10 posttransplantation during the post-irradiation platelet nadir. Further treatment led to platelet counts higher than normal 18 days posttransplantation when control animals had just attained normal platelet counts. IL-11 can accelerate the recovery of the peripheral blood leukocytes, mainly neutrophils, and platelets in transplant mice, effects that may be clinically useful in future applications for BM transplantation and chemotherapy-related cytopenias. PMID- 8417799 TI - Nomenclature of exogenous hemostatic factors. PMID- 8417800 TI - Transcriptional and posttranscriptional regulation of the interleukin-4 and interleukin-3 genes in human T cells. AB - Human T cells were studied with regard to the regulation of interleukin-4 (IL-4) and IL-3 gene expression. IL-4 and IL-3 mRNA were undetectable in unstimulated T cells. On activation with the lectin concanavalin A (Con A), both IL-4 and IL-3 mRNA were expressed. Accumulation of IL-4 mRNA peaked after 6 to 12 hours, whereas IL-3 mRNA levels peaked after 3 to 6 hours of stimulation with Con A. Nuclear run-on assays showed a low constitutive transcription for both genes. The transcription rates were increased by Con A resulting in a peak for IL-4 after 1 hour (30% increase) and for IL-3 after 3 hours (40% increase) of Con A treatment. mRNA stability studies demonstrated that on activation with Con A both messages decayed with a half-life of approximately 90 minutes. No IL-4 or IL-3 mRNA expression was induced by the protein kinase C activator phorbol myristate acetate (PMA). However, PMA augmented the Con A-induced IL-4 and IL-3 mRNA accumulation. This was shown to be mediated at posttranscriptional level by a large increase in the stability of both messages (t 1/2 > 3 hours). The transcription rate of both genes was also enhanced by Con A+PMA and reached peak levels for IL-4 after 1 hour (90% increase) and for IL-3 after 3 hours (70% increase) of stimulation. Furthermore, it appeared that the induction of IL-4 mRNA was dependent on protein synthesis because cycloheximide (CHX) blocked the Con A- and Con A+PMA-induced expression of IL-4 mRNA. In contrast, CHX inhibited, but failed to completely block, the Con A- and Con A+PMA-induced IL-3 mRNA expression, whereas the expression of both genes was completely blocked by cyclosporine A. With regard to the secretion of IL-4 protein it was shown that it closely follows the accumulation of IL-4 mRNA. Taken together, the data show that expression of the IL-4 and IL-3 genes in human T cells is controlled by different activation pathways that affect the gene regulation at transcriptional and posttranscriptional levels. PMID- 8417801 TI - Human platelets secrete chemotactic activity for eosinophils. AB - Thrombin-stimulated platelets liberate factors that induce chemotaxis of eosinophils and raise their cytosolic Ca2+ content ([Ca2+]i). The sources of this activity are the dense- and alpha-granules because inhibition of prostaglandin endoperoxide/thromboxane A2 formation and the platelet-activating factor receptor antagonist WEB 2086 have no effect. Platelets from patients with Storage-Pool Deficiency show about 60% of the normal chemotactic activity with little effect on [Ca2+]i, whereas completely degranulated platelets fail to affect eosinophils. In concentrations secreted by the platelets, adenosine diphosphate (ADP), and platelet factor 4 have no effect, whereas adenosine triphosphate (ATP) induces a strong chemotactic response and increases [Ca2+]i. However, apart from ATP other modulating factors must be involved as platelet releasates induce more chemotaxis than ATP alone. Thus, platelets secrete factors that activate eosinophils and may contribute to inflammatory and allergic processes. PMID- 8417802 TI - Clearance of human native, proteinase-complexed, and proteolytically inactivated C1-inhibitor in rats. AB - C1-inhibitor is the only known inhibitor of the classical pathway of complement and the major inhibitor of the contact pathway of coagulation. Like other serine proteinase inhibitors, C1-inhibitor can exist in three conformations, ie, the native, the proteinase-complexed, and the proteolytically inactivated form. Here we studied the plasma elimination kinetics of these three forms of human C1 inhibitor in rats. The clearance of the complexed form of C1-inhibitor appeared to be the most rapid and depended in part on the proteinase involved (observed plasma t1/2 was 20 minutes for C1s-C1-inhibitor, 32 minutes for kallikrein-C1 inhibitor, and 47 minutes for beta XIIa-C1-inhibitor), whereas that of native C1 inhibitor was the slowest (observed plasma t1/2 4.5 hours). Inactivated C1 inhibitor was cleared with an apparent plasma t1/2 of 1.6 hours. Thus, the short plasma t1/2 of complexed relative to native C1-inhibitor explains why in patients only low concentrations of C1-inhibitor complexes may be observed despite activation of the contact and/or complement systems. PMID- 8417803 TI - Arg-Gly-Asp-dependent occupancy of GPIIb/IIIa by applaggin: evidence for internalization and cycling of a platelet integrin. AB - Using indirect immunofluorescence microscopy we examined the distribution and cycling of GPIIb/IIIa after binding to applaggin, a high-affinity Arg-Gly-Asp (RGD)--containing ligand. Resting, unfixed platelets were incubated with applaggin for 30 minutes at 37 degrees C, and bound applaggin was detected by an affinity-purified rabbit anti-applaggin antibody. Examination of intact cells showed a rim pattern for applaggin, consistent with its binding to the platelet surface. Staining of Triton X-100--permeabilized cells showed an intracellular pool of applaggin. Competition of applaggin binding by either AP-2, an anti GPIIb/IIIa monoclonal antibody (MoAb) that blocks fibrinogen binding, or the synthetic peptide RGDW eliminated both surface and intracellular staining, indicating that applaggin is binding to GPIIb/IIIa in an RGD-dependent manner. Inhibition of platelet activation by PGE1 and theophylline had no effect on the observed staining patterns, indicating that cellular activation is not required for surface binding and subsequent internalization. To evaluate whether occupancy of functional binding sites on GPIIb/IIIa is required for internalization, we used mAb15, an anti-GPIIIa antibody that neither blocks fibrinogen binding nor induces the expression of ligand-induced binding sites on GPIIb/IIIa. In these studies mAb15 was internalized in a manner analogous to both AP-2 and applaggin, showing that occupancy of the RGD binding site is not required to initiate receptor internalization. To estimate the size of the newly internalized pool of applaggin, 125I-applaggin--binding studies were performed. Displacement of bound 125I-applaggin by excess unlabeled applaggin or EDTA showed that at least 17% of bound applaggin was nondisplaceable when binding was performed under conditions permitting membrane flow and internalization. These data indicate that GPIIb/IIIa is internalized in unstimulated platelets independent of cellular activation or occupancy of the functional binding site(s) of GPIIb/IIIa by RGD-containing ligands. Thus, internalization of GPIIb/IIIa may represent a mechanism by which the surface expression of this adhesion receptor is regulated. PMID- 8417804 TI - Vitamin K1 metabolism and the production of des-carboxy prothrombin and protein C in the term and premature neonate. AB - This study investigated the incidences of undercarboxylated (protein induced by vitamin K absence: PIVKA) prothrombin and protein C in 496 neonates across a wide range of gestational ages. These findings are related to vitamin K1 levels (an indicator of cofactor availability) and vitamin K1 epoxide levels (a measure of the efficiency of the hepatic vitamin K cycle). PIVKA protein C was present in at least trace amounts in 27% of infants; whereas, PIVKA prothrombin was present in 7% of infants. PIVKA prothrombin and protein C were present at high plasma concentrations in 2% to 3% of term and preterm neonates and both PIVKA protein C and prothrombin increased with gestational age. Despite elevated plasma concentrations of PIVKA protein C and diminished levels of normally carboxylated protein C, clinical thrombosis was not observed. The mean (+/- SD) vitamin K1 level in the study population was 0.009 +/- 0.02 nmol/L (adult reference interval: 0.3 to 2.6 nmol/L) with no clear relationship between vitamin K1 levels and production of PIVKA protein C or prothrombin. By comparison with adults, the epoxide form of the vitamin comprised an abnormally high proportion of total vitamin K1; this suggests possible inefficiencies in hepatic reductase cycling. PMID- 8417805 TI - Three new monoclonal antibodies that define a unique antigen associated with prolymphocytic leukemia/non-Hodgkin's lymphoma and are effectively internalized after binding to the cell surface antigen. AB - Prolymphocytic leukemia (PLL) is closely related to chronic lymphocytic leukemia (CLL), but present with distinctive clinical/laboratory features and associated with much worse prognosis. In this study, we generated three new IgG1-kappa monoclonal antibodies (MoAbs), termed SN8, SN8a and SN8b, by use of an unconventional approach, ie, by using an isolated B PLL antigen preparation to immunize mice. These MoAbs, particularly SN8, showed a highly selective reactivity to B PLL and B non-Hodgkin's lymphoma (NHL) among various human leukemia-lymphoma specimens tested; eg, SN8 was capable of effectively distinguishing B PLL from B CLL as well as from hairy cell leukemia (HCL) cell specimens. The cell surface antigen defined by the three MoAbs was determined to be a covalently linked heterodimeric glycoprotein complex (gp49/40) consisting of a 49,000 dalton (alpha-chain) and a 40,000-dalton component (beta-chain). Epitope comparison showed that the epitope defined by SN8 (SN8 epitope) is in close proximity to SN8a epitope but in a distant position from SN8b epitope. Western blot analysis showed that both SN8 and SN8a epitopes are on the beta-chain, but SN8b epitope was not detected on either the alpha- or the beta-chain of the reduced antigen in the same analysis. Binding of either SN8 or SN8b to the cell surface gp49/40 did not cause significant downregulation of the antigen expression whereas binding of SN8a to the antigen caused small (approximately 20%) decrease in the antigen expression. Among the various normal peripheral blood cells, only a subpopulation (6.0% to 24.2% among different specimens derived from different donors) of B cells reacted with the SN8 series MoAbs; these MoAbs showed no significant reactivity against T cells, granulocytes, monocytes, erythrocytes, and platelets. Minimal or no significant reactivity (0 to 2.6% among different specimens) was detected against normal bone marrow cells. Ricin A-chain conjugates of the three MoAbs are all strongly effective for specific killing of SN8 antigen-expressing leukemia cells in the absence of any potentiators; furthermore, the addition of 10 mmol/L NH4Cl, a potentiator, enhanced strongly the cytotoxic activities of the SN8, SN8a, and SN8b conjugates. Thus, each of the three MoAbs was effectively internalized after binding to the cell surface antigen. PMID- 8417806 TI - F reticulocyte response in sickle cell anemia treated with recombinant human erythropoietin: a double-blind study. AB - Studies on baboons and preliminary observations in three patients with sickle cell anemia (SS) suggested that high doses of pulse administered recombinant human erythropoietin (rHuEPO) stimulate F-reticulocyte production. We now report on the administration of rHuEPO in a double-blind format to ascertain frequency of response and potential precipitation of side effects. Ten patients were enrolled, but one was discontinued due to the indication of a blood transfusion. Of the other nine, five received rHuEPO in escalating doses (from 400 to 1,500 U per kg twice daily [BID] per week), alternating with a placebo, in blinded fashion. The second group, consisting of four patients, followed an identical protocol (except starting dose was 1,000 U/Kg, BID per week) and were iron supplemented during treatment. The criterion of response was a transient doubling (as a minimum) of the steady-state F-reticulocyte level. We found that none of the five patients in the first group responded to rHuEPO, and two of them became iron deficient, as judged by a significant decrease in ferritin. Of the second group, four patients responded with F-reticulocyte increases. In three patients, open label administration of rHuEPO confirmed the effect. We observed seven painful episodes during this study, two during the EPO administration and five during the placebo arm. Three patients were phlebotomized because the hemoglobin level increased 1.5 g/dL more than steady-state levels. Of the six patients followed-up by percent dense cell determinations, one exhibited increased levels during periods of the treatment, whereas the other five showed no change. No anti rHuEPO antibodies were detected. We conclude that rHuEPO can stimulate F reticulocyte response in some patients with sickle cell anemia, without apparent negative clinical side effects. The state of iron stores may be critical. Whether higher doses of rHuEPO and/or a different regimen might induce sustained F cells and fetal hemoglobin increases remains to be determined. PMID- 8417807 TI - Human T-lymphotropic virus type I tax regulates the expression of the human lymphotoxin gene. AB - Human T-lymphotropic virus type-I (HTLV-I)-infected T-cell lines constitutively produce high levels of lymphotoxin (LT). To analyze the mechanisms that lead to the expression of LT in HTLV-I-infected cell lines, we studied regulatory regions of the human LT promoter involved in the activation of the human LT gene. As determined by deletional analysis, sequences between +137 and -116 (relative to the transcription initiation site) are sufficient to direct expression of a reporter gene in the HTLV-I-infected cell line MT-2. Site-directed mutation of a of the single kappa B-like motif present in the LT promoter region (positions -99 to -89) completely abrogated LT promoter activity in MT-2 cells, suggesting that this site plays a critical role in the activation of the human LT gene. Transfection of LT constructs into HTLV-I-uninfected and -unstimulated Jurkat and U937 cell lines showed little to no activity of the LT promoter. Cotransfection of the same constructs with a tax expression plasmid into Jurkat cells led to detectable promoter activity, which could be significantly increased by stimulation of the cells with phorbol myristate acetate (PMA). Similarly, cotransfection of the LT promoter constructs and the tax expression plasmid into U937 cells led to significant promoter activity upon stimulation with PMA. These data suggest that HTLV-I tax is involved in the upregulation of LT gene expression in HTLV-I-infected cells. PMID- 8417808 TI - Chromosome mapping of murine susceptibility loci to liver carcinogenesis. AB - The validity of mouse liver tumors is controversial in the risk assessment of carcinogenicity of chemicals in humans, because mice used in carcinogenicity bioassays are genetically predisposed to liver tumors. The argument could be resolved once liver tumor susceptibility genes have been cloned and their role in liver tumor development elucidated. We performed a genetic linkage analysis to map murine liver tumor susceptibility genes, as a first step toward their identification. An F2 population of 87 urethane-treated male A/J x C3H/He mice was scored with 83 genetic markers. Three regions, localized on chromosomes 7, 8, and 12, were found to contain putative liver tumor susceptibility genes. PMID- 8417809 TI - A linkage analysis of D17S74 (CMM86) in thirty-five families with premenopausal bilateral breast cancer. AB - We report here results of a linkage analysis of a marker in 35 families in which the proband had premenopausal bilateral breast cancer. This group is of particular interest given their high family risk and the question of etiological heterogeneity. Probands were ascertained from cancer registries in Los Angeles County and Connecticut and major hospitals in Montreal and Quebec. Assuming no residual heterogeneity and summing lod scores over all families, we obtained strong evidence against tight linkage (e.g., lod score at theta = 0.000001 is 3.39). To address the issue of heterogeneity, we performed admixture and predivided sample analyses. Using an admixture model we were able to reject the hypothesis of no linkage versus that of linkage with homogeneity (P = 0.045). However, we were unable to reject the hypothesis of no linkage versus linkage with heterogeneity (P = 0.119) or to distinguish between linkage with homogeneity and linkage with heterogeneity (P = 0.500). Predivided sample analyses based upon age of onset, pathological characteristics, time between diagnoses of the breast cancers in each bilateral proband, and the span of ages at diagnoses within a family did not discriminate between apparently linked and unlinked families. PMID- 8417810 TI - Delta elk-1, a variant of elk-1, fails to interact with the serum response factor and binds to DNA with modulated specificity. AB - The ets oncogene superfamily codes for a family of transcriptional factors that are involved in gene regulation not only by autonomous DNA binding but also by indirect DNA binding through interaction with cellular factors. We have previously shown that a member of this superfamily, elk-1, is a sequence specific transcriptional activator, which forms a serum response factor (SRF) dependent ternary complex with serum response element (SRE) similar to p62TCF. We describe here an alternatively spliced variant of elk-1 named delta elk-1, which has lost the SRF interaction domain, negative regulatory DNA binding domain, and part of the elk-1 DNA binding domain. This variant elk-1 protein has lost the capacity to form a SRF dependent ternary complex with SRE and to activate fos transcription. Since this splice variant lacks part of the ets DNA binding domain, it binds to DNA with a specificity that is different from that of the full length elk-1 protein. Therefore differential splicing within the DNA binding and protein protein interaction domains of transcriptional factors can generate proteins with modulated DNA binding specificities and transcriptional regulation. Thus it is conceivable that variant elk-1 might function by competing for some of the elk-1 target sequences (like SRE) and thereby block the transcriptional activation of fos by SRF and elk-1. Alternately, variant elk-1 protein may be the repressor, recruited by the SRE bound SRF for c-fos repression, or it may have an altogether different function. Therefore, elk-1 appears to fall in the category of genes that encode activators and repressors through the mechanism of differential splicing. PMID- 8417811 TI - Inhibition of human platelet glycoprotein IIB/IIIA binding to fibrinogen by tumor cell membrane proteins. AB - Immunochemical and functional characteristics of tumor cell membrane proteins and human platelet glycoproteins were studied. Immunoblotting revealed that membrane proteins of a cultured breast tumor cell line (BT-20) had three protein bands, which were each recognized by monoclonal antibodies to human platelet glycoprotein Ib, IIb, and IIIa, suggesting some immunochemical similarities between the tumor cell membrane proteins and platelet glycoproteins. The monoclonal antibodies failed to bind to an extract of a lung tumor cell line (A549). Neither tumor extract induced platelet aggregation. However, tumor associated antigens isolated from the breast tumor cells markedly inhibited platelet glycoprotein IIb/IIIa binding to fibrinogen. In contrast, tumor associated antigens from the lung tumor cells had no effect. These results suggest that tumor cells which have immunological and/or structural similarities to platelets may affect hemostasis and coagulation in vivo. PMID- 8417812 TI - Molecular cloning of a complementary DNA encoding a prostate-specific membrane antigen. AB - Recently, a novel M(r) 100,000 prostate-specific membrane glycoprotein (PSM) has been detected by the prostate-specific monoclonal antibody 7E11-C5, raised against the human prostatic carcinoma cell line LNCaP. The PSM antigen is expressed exclusively by normal and neoplastic prostate cells and metastases. We now report the molecular cloning of a full-length 2.65-kilobase complementary DNA encoding the PSM antigen from a human LNCaP complementary DNA library by polymerase chain reaction using degenerate oligonucleotide primers. Analysis of the complementary DNA sequence has revealed that a portion of the coding region, from nucleotide 1250 to 1700, has 54% homology to the human transferrin receptor mRNA. The deduced polypeptide has a putative transmembrane domain enabling the delineation of intra- and extracellular portions of this antigen. In contrast to prostate-specific antigen and prostatic acid phosphatase which are secreted proteins, PSM as an integral membrane protein may prove to be effective as a target for imaging and cytotoxic targeting modalities. PMID- 8417813 TI - Preferential increase of glutathione S-transferase class alpha transcripts in cultured human hepatocytes by phenobarbital, 3-methylcholanthrene, and dithiolethiones. AB - In rodents, a diversity of compounds are able to protect against acute and chronic toxicities of various xenobiotics including carcinogens, at least in part through induction of drug-metabolizing enzymes including glutathione S transferase (GST) enzymes. We have posed the question as to whether or not these compounds also induce GSTs in human liver. Primary human hepatocyte cultures were exposed to phenobarbital, 3-methylcholanthrene, and two dithiolethiones [1,2 dithiole-3-thione and its 5-(2-pyrazinyl)-4-methyl derivative, oltipraz], and steady-state mRNA levels of GST classes alpha, mu, and pi were determined by Northern blot analysis. After 3 daily treatments, the two dithiolethiones were the most potent inducers; phenobarbital was also effective but to a lesser extent and 3-methylcholanthrene increased GST mRNA in only 2 of the 6 samples, although it stimulated cytochrome P-450 1A2 mRNA in all cell preparations. Whatever the compound only GSTA1 and/or A2 transcripts were induced. GST M1 mRNAs were not responsive or only slightly responsive, and GST P1 mRNAs, which were mostly undetectable in control cells, were not affected by treatment with any of the four chemicals. Large individual variations were observed in the level of induction of GST A1 and/or A2 mRNAs, and no sex difference could be demonstrated. These results clearly indicate that phenobarbital, 3-methylcholanthrene, and dithiolethiones are able to markedly increase mRNA levels of GST in human hepatocytes and that the GST alpha class is preferentially involved. PMID- 8417814 TI - Cyclin A expression in human hematological malignancies: a new marker of cell proliferation. AB - Several in vitro studies have shown that the cyclin A gene is expressed and plays an important role in both the S and G2-M phases of the cell cycle. We analyzed cells from the blood and bone marrow of patients with various, mostly neoplastic, hematological disorders to determine whether (a) the cyclin A protein level correlated with that of cyclin A RNA and (b) cell distribution among the different phases of the cell cycle correlated with cyclin A RNA expression. Thirty-eight patients were studied by means of dot blot and Western blot techniques for cyclin A RNA and protein accumulation, and 21 were also studied for cell cycle distribution by using flow cytometric analysis. Semiquantitative studies were based on densitometric computerized evaluation of dot and Western blots. There was a very strong positive correlation between cyclin A RNA and protein expression (r = 0.99; P < 0.00005), indicating that cyclin A accumulation is regulated in these cells at a transcriptional level. There was also a highly significant positive correlation between cyclin A RNA expression and the cumulative percentage of cells in S plus G2-M phase (r = 0.98; P < 0.00005). Therefore, this in vivo study shows that the expression of cyclin A RNA and protein in human hematological malignancies correlates with the percentage of cells in S plus G2-M phase and identifies cyclin A as a new potential cell proliferation index in oncology. PMID- 8417815 TI - Hyperphosphorylation of retinoblastoma protein and p53 by okadaic acid, a tumor promoter. AB - A potent tumor promoter, okadaic acid, induced hyperphosphorylation of tumor suppressor proteins, retinoblastoma protein and p53, by in vitro incubation with nuclei isolated from rat regenerating liver as well as by incubation with primary human fibroblasts. Most of the retinoblastoma protein migrated to a hyperphosphorylated position in electrophoresis. The phosphorylation of p53 was increased at a rate 8 times that in non-treated primary human fibroblasts. Hyperphosphorylation of tumor suppressor proteins, mediated through inhibition of protein phosphatases 1 and 2A, is involved in tumor promotion by okadaic acid. The significance of hyperphosphorylation of the retinoblastoma protein and p53 is discussed in relation to the regulation of the cell cycle. PMID- 8417816 TI - Ormaplatin sensitivity/resistance in human ovarian cancer cells made resistant to cisplatin. AB - The human ovarian cancer cell lines A2780 and A2780/CP70 were studied to investigate the cellular basis for their relative sensitivities to tetrachloro(DL trans)-1,2-diamminecyclohexaneplatinum(IV) (ormaplatin). Cells were exposed to ormaplatin for 1 h in all experiments. As assessed by colony formation assays, the A2780/CP70 cell line [50% inhibitory dose (IC50) = 3.6 microM] was 9.5-fold more resistant to ormaplatin than the A2780 cell line [IC50 = 0.38 microM]. For cisplatin, the IC50 doses were 40 and 3 microM, respectively. Both cell lines were treated with ormaplatin at doses ranging from 0.10 to 40 microM, for the purpose of studying drug accumulation and efflux, and DNA adduct formation and repair. When these cell lines were treated at their respective IC50 doses, drug accumulation was greater in the resistant cells. When treated at equal microM doses, the sensitive cells formed 8-fold more DNA adduct than the resistant cells. When cells were treated with ormaplatin so as to achieve equivalent levels of platinum-DNA modification, sensitive cells removed 53% of the platinum-DNA damage in the first 6 h after drug exposure, compared to 68% in the resistant cells. We conclude that in human ovarian cancer cells made resistant to cisplatin, there is moderate cross-resistance to ormaplatin. This cross resistance is not explained by differences in drug accumulation but is associated with reduced platinum-DNA adduct formation, which may be attributable in part to cytosolic inactivation of drug. PMID- 8417817 TI - Mechanism of the protective effect of supplemental dietary calcium on cytolytic activity of fecal water. AB - Dietary calcium supplementation inhibits hyperproliferation of rectal epithelium, possibly by precipitating luminal surfactants and thus preventing their cell damaging effects. Therefore, we studied the effects of supplemental dietary calcium (35.5 mmol/day) on composition and cytolytic activity of fecal water and on the release of the epithelial marker alkaline phosphatase in 12 healthy volunteers. Fecal water was isolated by low-speed centrifugation. Cytolytic activity was determined as lysis of human erythrocytes by fecal water. Intestinal alkaline phosphatase activity in fecal water was measured with the use of the uncompetitive inhibitor L-phenylalanine. Supplemental calcium increased soluble calcium and decreased soluble P(i). The logarithm of the concentration product of calcium and phosphate was linearly dependent on pH. These observations indicate formation of insoluble calcium phosphate. Supplemental calcium did not alter the total bile acid concentration in fecal water but significantly decreased the ratio of more hydrophobic to more hydrophilic bile acids from 3.3 to 2.3. Calcium also significantly decreased the concentration of fatty acids (from 2.9 to 2.1 mM). Consistent with these decreases in hydrophobic surfactants, calcium decreased the cytolytic activity of fecal water from 47 +/- 9 to 27 +/- 8% (n = 12, P < 0.05). Analogous to the decrease in cytolytic activity, the release of the epithelial marker alkaline phosphatase was also lowered by supplemental calcium. We conclude that supplemental dietary calcium decreases luminal cytotoxic surfactant concentrations and thus inhibits luminal cytolytic activity and the release of the epithelial marker alkaline phosphatase as an indicator of intestinal epitheliolysis. This mechanism may explain how dietary calcium could decrease epithelial cell proliferation. PMID- 8417818 TI - Neopterin and prognosis in patients with adenocarcinoma of the colon. AB - Concentrations of neopterin, a sensitive indicator for the activation of cellular immunity, were measured in urine samples of 44 patients with adenocarcinoma of the colon at diagnosis. To judge the relative predictive value of neopterin concentrations, other routine clinical and laboratory variables were concomitantly determined. The patients were then followed up to 10 yr, and the abilities of all variables to predict death from cancer were assessed. Neopterin concentrations were not correlated with either tumor stage or Dukes' stage. In univariate analyses using the product-limit approach, four variables were significant indicators of poor prognosis: presence of distant metastases (P = 0.0001); high Dukes' stage (P = 0.0009); high urinary neopterin concentration (P = 0.0034); and advanced stage (P = 0.030). Presence versus absence of lymph node metastases was not associated with prognosis. Multivariate survival analyses by the proportional hazards technique demonstrated that neopterin provided statistically independent predictive information in addition to either presence versus absence of distant metastases or Dukes' stage. When neopterin and tumor stage were investigated for joint prediction, stage failed to be included in the model. Thus, neopterin concentrations provide valuable and statistically independent prognostic information in patients with adenocarcinoma of the colon. PMID- 8417819 TI - Phase I study of the oral nonsteroidal aromatase inhibitor CGS 20267 in postmenopausal patients with advanced breast cancer. AB - A phase I study was performed of CGS 20267, an oral nonsteroidal, highly potent, and selective aromatase inhibitor, in 21 postmenopausal patients with advanced breast cancer. The patients were recruited in 3 successive groups of 7, receiving 0.1, 0.5, and 2.5 mg p.o./day, respectively. All patients had received at least one prior endocrine treatment (range, 1-4), and six patients had received prior chemotherapy. The treatment was very well tolerated, and no toxicity was seen at any of the three doses. There was a statistically significant suppression of estradiol (E2) and estrone (E1) levels by 74% and 79% from baseline levels, respectively (P < 0.0001). Suppression occurred in all three patient groups, with many patients having serum concentrations of estradiol and estrone, which were below the limit of detection of the assays (3 and 10 pM, respectively), which corresponds to a maximum measurable estrogen suppression of 86%. CGS 20267 had no significant effect on serum levels of follicle-stimulating hormone, luteinizing hormone, thyroid-stimulating hormone, cortisol, 17 alpha-hydroxyprogesterone, androstenedione, and aldosterone. Seven (33%, 95% confidence interval, 15-57%) of the 21 patients have responded to treatment (one complete remission, 6 partial remissions according to criteria of the Union Internationale contre le Cancer), and 6 are still responding to CGS 20267 (duration of response; 4+, 6+, 6+, 9+, 9, 12+, and 12+ months). Five have had stable disease for more than 3 months, and 9 had progressive disease. These results suggest that CGS 20267 is a very potent and specific aromatase inhibitor, and phase II studies are now required to confirm its clinical efficacy. PMID- 8417820 TI - Comparative dual label study of first and second generation antitumor-associated glycoprotein-72 monoclonal antibodies in colorectal cancer patients. AB - Radiolabeled first-generation anti-tumor-associated glycoprotein-72 (TAG-72) monoclonal antibody (MAb), B72.3, has proven useful in detecting primary and secondary colorectal carcinoma. It has been anticipated that the development of second-generation, higher affinity, anti-TAG-72 MAbs, CC49 and CC83, would be of greater use in cancer detection and of value in radioimmunotherapy of human cancer. We compared the pharmacokinetics, biodistribution, and immune responses of 131I-labeled CC49 and CC83 to 125I-labeled B72.3 by preoperatively coninjecting dual-labeled MAbs into 16 colorectal cancer patients. The imaging properties of radiolabeled CC49 and CC83 were also assessed. Pharmacokinetics of all three MAbs were identical, and there were no differences in the uptake of any of three MAbs in tumor and normal tissues. Maximum tumor uptake was 0.0041% of the injected dose/g for 125I-B72.3, 0.0024% for 131I-CC49, and 0.0029% for 131I CC83. Radiolabeled CC49 and CC83 detected most known tumor sites on scintigrams without any clear advantage for either MAb. Nonspecific splenic and testicular uptake was frequently observed. Anti-idiotypic human anti-mouse antibody responses were seen more frequently with B72.3 than with CC49 or CC83. We conclude that higher affinity, radiolabeled anti-TAG-72 MAbs can detect colorectal cancer but do not penetrate these tumors more effectively than B72.3. Improvements in tumor detection and radioimmunotherapeutic strategies will likely require the administration of smaller fragments of MAb molecules or novel delivery systems rather than the continued development of higher affinity MAbs. PMID- 8417821 TI - Expansion of the epithelial cell proliferative compartment and frequency of adenomatous polyps in the colon correlate with the strength of family history of colorectal cancer. AB - Expansion of the proliferative compartment of epithelial cells in colonic crypts and colonic adenomas have been described as phenotypic precursors to colon cancer in individuals affected with hereditary or sporadic colon cancer. This study measured the size of the proliferative compartment in colonic crypts and the frequency of adenomas in asymptomatic members of families having sporadic colorectal cancer. The subjects were divided into 2 groups according to the frequency of colorectal cancer in their families. A shift of the compartment of proliferating epithelial cells toward the lumenal surface of colonic crypts was seen in the group of subjects with a stronger family history of colorectal cancer, with significant differences in the numbers of proliferative cells in the upper and the lower crypt compartments (P < 0.05) and in the fraction of proliferative cells at the highest compartment at the lumenal surface of the crypts (P < 0.05). Cell proliferation patterns in normal-appearing mucosa of the 2 groups revealed no difference in whole crypt [3H]thymidine labeling index. Colonoscopic examination of the 56 subjects revealed an overall prevalence of adenomas of 21%; when stratified by frequency of colorectal cancer in their families, 3 of 22 subjects (14%) with a weaker family history had adenomas, while 9 of 34 (26%) with a stronger family history had adenomas. Thus, parallel abnormalities of colonic epithelial cell proliferation and neoplasia were seen in individuals with a family history of colorectal cancer, both of which were more pronounced with increasing strength of family history. This observation provides further evidence of relationships among these factors in the etiology of "sporadic" colorectal cancer. PMID- 8417822 TI - Induction of jun gene family members by transforming growth factor alpha but not 17 beta-estradiol in human breast cancer cells. AB - To investigate whether estrogen treatment of hormone-responsive human breast cancer cells was associated with activation of members of the jun family of immediate early response genes, the expression of these oncogenes in human breast cancer cells was examined. 17 beta-Estradiol had little effect on expression of c jun, jun B, jun D, or c-fos mRNA by MCF-7 cells over 12 h, although it stimulated c-myc expression 4-fold within 30 min. In contrast, several peptide growth factors, including transforming growth factor-alpha (TGF-alpha), rapidly and transiently induced expression of c-jun, jun B, and c-fos mRNA 4- to 10-fold over control. A similar pattern of expression was seen in two other estrogen responsive human breast cancer cell lines, ZR-75-1 and T47D. Inhibition of protein synthesis by cycloheximide did not abrogate induction of c-jun or jun B mRNA by TGF-alpha in MCF-7 cells, suggesting that new protein synthesis was not required. In addition, nuclear runoff transcription analysis demonstrated that increased expression of c-jun and jun B mRNA after TGF-alpha treatment of MCF-7 cells was regulated at least in part at the transcriptional level. Chronic exposure of MCF-7 cells to 17 beta-estradiol for 24-48 h was associated with decreased expression of jun B mRNA only, while similar treatment with TGF-alpha did not change mRNA expression of any jun family member. Thus, expression of jun family members is induced by peptide growth factors like TGF-alpha but not 17 beta-estradiol in human breast cancer cells. These results suggest that these nuclear protooncogenes play different roles in modulating gene expression by MCF 7 cells after exposure to TGF-alpha or 17 beta-estradiol. PMID- 8417823 TI - Inhibition of estrone sulfatase activity by estrone-3-methylthiophosphonate: a potential therapeutic agent in breast cancer. AB - Many breast tumors are hormone dependent, and there is evidence that hydrolysis of estrone sulfate (E1S) to estrone, by estrone sulfatase, is an important source of the estrogen which is found in tumors. In this study, we have developed a novel pathway for the synthesis of estrone-3-methylthiophosphonate (E1-3-MTP) and examined its ability to inhibit estrone sulfatase activity in MCF-7 breast cancer cells and human placental and breast tumor preparations. In MCF-7 breast cancer cells, E1-3-MTP, 100 nM and 10 microM, inhibited estrone sulfatase activity by 52 and > 98%, respectively. The apparent Km and Vmax for E1S were 4.8 microM and 148 pmol/min/mg for placental and 16.9 microM and 38 pmol/min/mg for breast tumor preparations. Kinetic studies revealed that E1-3-MTP inhibited estrone sulfatase in a competitive manner with the Ki values for placental and tumor preparations being 14.6 and 32.8 microM, respectively. A comparison of the metabolism of [3H]E1S and [3H]E1-3-MTP by human placenta or rat liver revealed that, whereas 50 60% of [3H]E1S was converted to [3H]estrone, < 3% of [3H]E1-3-MTP was hydrolyzed. The development of an efficient inhibitor of estrone sulfatase, which is resistant to metabolism, will allow the importance of the estrone sulfatase pathway of estrogen formation in breast tumors to be assessed and such an inhibitor may have considerable potential as a therapeutic agent. PMID- 8417824 TI - Uptake and storage of m-iodobenzylguanidine are frequent neuronal functions of human neuroblastoma cell lines. AB - The mechanisms of the uptake and release of m-iodobenzylguanidine (MIGB) have been studied in 5 neuroblastoma (NB) cell lines and in 4 clonal NB sublines with a homogeneous phenotype. A specific uptake system for MIBG was found in 8 of 9 NB cell lines or subpopulations. The uptake was characterized by temperature dependency, high affinity, saturability, sodium dependency, and imipramine sensitivity. The majority of NB cell lines that possessed a specific uptake system for MIBG were also able to efficiently store the incorporated drug. However, 3 NB cell lines were identified without the ability to retain high levels of MIBG, despite the presence of a specific uptake system. We also report that a clonal subline, SH-EP1, which has a nonneuroblastic phenotype, failed both MIBG uptake and retention. Conversely, the parental cell line, SK-N-SH, and the neuroblastic subline SH-SY5Y possessed both a specific uptake system and the ability to store MIBG. In addition, the induction of neuronal differentiation with retinoic acid increased the velocity of uptake and the storage efficiency for MIBG in the clonal subline SH-SY5Y. We conclude that MIBG uptake and storage should be considered to be frequent but independent neuronal functions of human NB cells. PMID- 8417825 TI - Antitumor effect of interleukin 1 alpha in combination with hyperthermia. AB - The combined effects of interleukin-1 alpha (IL-1 alpha) and hyperthermia on SCK tumors grown in the legs of A/J mice were investigated. When the host mice were given i.p. injections of 25 micrograms/kg IL-1 alpha, the tumor blood perfusion, as measured with the 86Rb uptake method, significantly declined, reaching minimum blood perfusion in 3-5 h. Although the tumor blood perfusion started to rise thereafter, the recovery was still incomplete 1 day later. Hyperthermia at 42.5 degrees C or 43.5 degrees C for 1 h also caused a marked decline in tumor blood perfusion. When the tumors were heated 5 h after an i.p. injection of IL-1 alpha at 25 micrograms/kg, at which time the tumor blood perfusion was low, the tumor blood perfusion decreased further. The heating of tumors at 42.5 degrees C or 43 degrees C for 1 h significantly reduced the clonogenic cell number in the tumors, delayed the tumor growth, and prolonged the survival time of host animals. The antitumor effect of 25 micrograms/kg IL-1 alpha alone, as judged from the clonogenic cell number, tumor growth delay, and host survival, was smaller than that of hyperthermia. When the host mice were treated with IL-1 alpha and the tumors were heated 5 h later, the decline in the clonogenic tumor cell number, tumor growth delay, and prolongation of host survival were significantly greater than those produced by either treatment alone. It was concluded that the prior reduction of blood perfusion by IL-1 alpha potentiated the antitumor effect of subsequent heatings. PMID- 8417826 TI - Interstitial chemotherapy of the 9L gliosarcoma: controlled release polymers for drug delivery in the brain. AB - The administration of drugs directly into the central nervous system using polymers as drug carriers may improve the treatment of malignant brain tumors. In this study, the effect of the interstitial, localized delivery of 1,3-bis(2 chloroethyl)-1-nitrosourea (BCNU) incorporated into controlled release polymers implanted adjacent to the 9L gliosarcoma was assessed in s.c. and intracranial (i.c.) models. In the s.c. experiment, the 9L gliosarcoma was implanted in the flank of rats and subsequently treated with BCNU either (a) delivered in controlled release polymers inserted adjacent to the tumor or (b) administered systemically by i.p. injections or by controlled release polymers inserted at a site distant from the tumor. The interstitial release of BCNU adjacent to the tumor in the flank resulted in a significant tumor growth delay of 16.3 days, as compared to a growth delay of 9.3 and 11.2 days obtained with the systemic administration of BCNU. In the i.c. experiment, the 9L gliosarcoma was implanted in the brain of Fischer 344 rats and treated either (a) with controlled release polymers containing BCNU inserted into the brain or (b) with the systemic i.p. administration of BCNU. The interstitial release of BCNU in the brain resulted in a significant 5.4- to 7.3-fold increased survival, compared with a 2.4-fold increased survival after the systemic administration of the same dose of BCNU. The two groups with i.c. tumors treated interstitially had 17 and 42% cures, but no long-term cures were obtained in the group treated with systemic therapy. The localized, controlled delivery of chemotherapeutic agents in the s.c. tissues and in the brain via polymeric carriers may be more effective than standard systemic chemotherapy. This approach could be used to deliver a wide variety of agents into the central nervous system to treat diverse neuropathological conditions which remain refractory to systemic therapy. PMID- 8417827 TI - BR96 sFv-PE40, a potent single-chain immunotoxin that selectively kills carcinoma cells. AB - We have constructed a single-chain immunotoxin composed of the carcinoma-reactive antibody BR96 and a truncated form of Pseudomonas exotoxin. The chimeric molecule, BR96 sFv-PE40, was expressed in Escherichia coli and localized to the inclusion bodies. We purified and identified two species of BR96 sFv-PE40, monomers and aggregates. The monomeric form was able to bind well to the BR96 antigen, a Lewisy-related antigen, while the aggregate was not. The binding affinity of the monomeric recombinant immunotoxin was 5-fold less than intact BR96 IgG, and its specificity for the BR96 antigen was confirmed by competition analysis. Monomeric BR96 sFv-PE40 was found to be extremely cytotoxic against cancer cells displaying the BR96 antigen. The cytotoxicity of the fusion protein correlates directly with antigen density on the tumor cell lines tested. The breast carcinoma cell line MCF-7, which has the highest density of BR96 antigen, was the most sensitive to BR96 sFv-PE40, with a concentration producing 50% protein synthesis inhibition of 5 pM. BR96 sFv-PE40 was found to have a t1/2 in serum of 28.5 min in athymic mice, compared to that of the chemical conjugate, chiBR96-LysPE40, which was 54 min. These data indicate that the single-chain immunotoxin BR96 sFv-PE40 is a potent inhibitor of protein synthesis in target cell lines and may be an effective agent for the treatment of cancer. PMID- 8417828 TI - Immunotoxins made with a recombinant form of Pseudomonas exotoxin A that do not require proteolysis for activity. AB - We used recombinant DNA technology to construct a mutant form of Pseudomonas exotoxin A (PE) called cysPE35 that contains amino acids 280-364 and 381-613 of PE. cysPE35 begins at the native PE proteolytic cleavage site and contains a single cysteine residue at position 287 that can be used to conjugate the toxin to monoclonal antibodies (MAbs). Unlike immunotoxins containing larger mutant forms of PE, such as PE40 or PE38, immunotoxins containing cysPE35 linked through a disulfide bond do not require proteolysis to generate a toxin fragment able to translocate to the cytosol. cysPE35 was conjugated to several MAbs and their activities were studied in vitro and in vivo. The concentration of toxin that inhibited protein synthesis as measured by a decrease in [3H]leucine incorporation of 50% of cysPE35 conjugated through a disulfide bond to the MAb HB21, which targets the human transferrin receptor, was 1 ng/ml on A431 cells. The MAb HB21 conjugated through a thioether bond to cysPE35 was much less active (concentration of toxin that inhibited protein synthesis as measured by a decrease in [3H]leucine incorporation of 50%, 200 ng/ml). An immunotoxin containing PE38 conjugated through either a disulfide or thioether bond to the MAb HB21 had a concentration of toxin that inhibited protein synthesis as measured by a decrease in [3H]leucine incorporation of 50% of 5 ng/ml, indicating that proteolysis of PE38 may be rate limiting in the action of these immunotoxins. Two other MAbs, LL2 and B3, were also conjugated through a disulfide bond to cysPE35. Both immunotoxins were also more active against cultured cells than conjugates using PE38 or PE40, and caused complete regression of human tumor xenografts growing in nude mice. In conclusion, we have constructed a mutant form of PE which must be coupled to MAbs through a disulfide bond to produce fully active immunotoxins that do not require proteolysis to generate a toxin fragment able to reach the cell cytosol. PMID- 8417829 TI - A mechanism for neutrophil-mediated lysis of human neuroblastoma cells. AB - Neutrophils mediate the lysis of human neuroblastoma cells coated with human/mouse chimeric anti-GD2 ganglioside antibody ch14.18. This study examined the mechanism(s) by which this occurs. Neutrophil degranulation was found to be a required step for lysis, since release of granular enzymes from neutrophils correlated with the lysis of antibody-coated neuroblastoma cells. In addition, agents which block degranulation specifically inhibited this process. Antibody dependent lysis of neuroblastoma cells was enhanced by exposing neutrophils to granulocyte-macrophage colony stimulatory factor. An increased release of lytic granular molecules was found to be responsible for this lymphokine-mediated phenomenon. Among the molecules released from neutrophil granules that were shown to be involved in neuroblastoma cell lysis were defensins, M(r) 3000-4000 neutrophil granular proteins which are known to bind and permeabilize tumor cells. In addition, cathepsin-G, a neutrophil granular protease, was demonstrated for the first time to mediate the lysis of human neuroblastoma cells. The enzymatic activity of cathepsin-G was found to be required for the lysis of these tumor cells, since phenylmethylsulfonyl fluoride blocks the lytic ability of this protein. PMID- 8417830 TI - Motility of human carcinoma cells in response to thrombospondin: relationship to metastatic potential and thrombospondin structural domains. AB - The interactions of tumorigenic cells with the extracellular matrix play a critical role in the establishment of metastases. Thrombospondin (TSP) is prominent at sites of tissue injury and promotes the attachment, spreading, and motility of several cell types. We have investigated the relationship between human carcinoma cell metastatic potential and TSP-mediated cell motility by comparing highly metastatic 11B carcinoma cells with a nonmetastatic counterpart, 22B carcinoma cells. 11B cells demonstrated motility in response to soluble TSP with a maximal effect observed at 1 microM TSP. Checkerboard analysis indicated that motility was directional with a significant chemokinetic component. Monoclonal antibody C6.7, specific for the distal COOH terminus of TSP, inhibited chemotaxis by 60%. Studies with TSP fragments demonstrated that the M(r) 140,000 COOH-terminal domain (140K) supported chemotaxis to the same extent as intact TSP. The NH2-terminal heparin-binding domain was ineffective in stimulating chemotaxis. Substrate-bound TSP also elicited 11B cell motility with a maximal response at 100 nM TSP. Directionality of this response was confirmed by checkerboard analysis. Interestingly, as in chemotaxis, haptotaxis was mediated exclusively by 140K as demonstrated by TSP fragment studies and inhibition with monoclonal antibody C6.7. Therefore, 140K appeared to mediate both chemotaxis and haptotaxis. Compared with 11B cells, 22B carcinoma cells are nonmetastatic and synthesize and secrete low levels of TSP. Immunoprecipitation and Northern blot analysis confirmed that 11B cells expressed much higher levels of TSP than 22B cells. Although 22B cells are able to attach to TSP, they did not exhibit either chemotaxis or haptotaxis in response to TSP or TSP fragments. Similarly, an antisense TSP cell line responded poorly in chemotaxis assays to TSP and 140K. These data suggest that the ability of metastatic cells in vivo to establish secondary sites of proliferation may be related to their ability to migrate in response to extracellular matrix proteins such as TSP incorporated into basement membranes or interstitial matrices. PMID- 8417831 TI - Human mesothelioma cells produce factors that stimulate the production of hyaluronan by mesothelial cells and fibroblasts. AB - The hyaluronan production by three human malignant mesothelioma cell lines and nine primary human mesothelial cell types was determined. The mesothelioma cell lines produced only minute amounts of hyaluronan (less than 0.1 microgram/10(6) cells/48 h) whereas mesothelial cells synthesized large quantities of hyaluronan (10-72 micrograms/10(6) cells/48 h). Conditioned media from the mesothelioma cell lines were investigated for their ability to stimulate hyaluronan production by fibroblasts and mesothelial cells in vitro, and in all cases stimulatory effects were found. The factor(s) in the conditioned medium of the mesothelioma cell line Mero-25 that were responsible for hyaluronan stimulation were heat stable and partially trypsin resistant. The stimulatory activity was partially inhibited by an antiserum against platelet-derived growth factor and basic fibroblast growth factor. Our data suggest that the increased hyaluronan synthesis seen in patients with mesothelioma is due to the release of factors from mesothelioma cells that stimulate other cells to produce hyaluronan. PMID- 8417832 TI - The A/J mouse lung as a model for developing new chemointervention strategies. AB - The use of the A/J mouse lung as a model for developing new chemo-intervention strategies was investigated by first inducing lung tumors with a single dose of 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanone. Lungs were then staged for tumor development and intervention therapy was initiated 42 weeks after carcinogen treatment. At this time point, an average of 7 pulmonary lesions were present on a standard histological section and the relative frequency of lesions was distributed as alveolar hyperplasias (38%), adenomas (40%), and adenocarcinomas (22%). Mice were treated for 4 or 8 weeks with cis-platinum alone or in combination with either indomethacin, an inhibitor of prostaglandin synthesis, metoclopramide, an inducer of poly(ADP) ribosylation, or nifedipine, a calcium channel blocker. The effect of indomethacin, metoclopramide, and nifedipine on tumor growth was also determined. The most dramatic effects were observed in lungs from mice treated for 8 weeks. cis-Platinum treatment caused a 37% reduction in the size of carcinomas, while tumor mass was reduced by 50 to 60% with cis-platinum in combination with metoclopramide and/or indomethacin. The inclusion of indomethacin therapy in conjunction with cis-platinum significantly enhanced the effectiveness of cis-platinum for inhibiting the growth of adenocarcinomas. In contrast, nifedipine appeared to ameliorate any of the inhibitory growth effects seen with cis-platinum treatment. Although none of the therapeutic combinations affected the size of adenomas, morphological differences were observed among treatment groups. A moderate to marked decrease in cytoplasm was observed in adenomas from mice treated with cis-platinum in combination with indomethacin or metoclopramide, cis-platinum plus metoclopramide and indomethacin, or metoclopramide plus indomethacin. Taken together, the results from these studies demonstrate that the A/J mouse lung can be used as a model to study the effectiveness of new intervention therapies for controlling malignant tumor growth. This model should also be applicable for studying the effectiveness of cancer prevention therapies on the progression of pulmonary hyperplasia. PMID- 8417833 TI - Expression of the metalloproteinase matrilysin in DU-145 cells increases their invasive potential in severe combined immunodeficient mice. AB - Human prostate cancer displays a high degree of variability in its rate of spread, which could be due largely to differences in the invasive potential of the tumor cells. The degradation of the basal lamina and stromal extracellular matrix is mediated in part by the secretion of matrix metalloproteinases (MMPs). Matrilysin (PUMP-1, MMP-7) and gelatinase A (M(r) 72,000 type IV collagenase, MMP 2) have been shown to be overexpressed in prostate carcinoma. We have expressed the single MMP matrilysin in the tumorigenic but nonmetastatic human prostate tumor cell line DU-145 to determine if matrilysin has a functional role in prostate tumor cell invasion. DU-145 cells expressing matrilysin were significantly more invasive than vector-only transfected cell lines as assayed by a severe combined immunodeficient mouse model of tumor cell invasion. Vector-only transfected DU-145 cells injected i.p. into severe combined immunodeficient mice invaded the diaphragm in only 1 of 9 mice (11%), whereas matrilysin-transfected DU-145 cells invaded the diaphragm in 12 of 18 mice (66%). The difference between the controls and matrilysin-transfected cells was statistically significant (P < 0.006). These results suggest a functional role for matrilysin in the initial invasion of prostate cancer through the epithelial basal lamina and into the surrounding stroma. PMID- 8417834 TI - Melanoma cells selected for adhesion to laminin peptides have different malignant properties. AB - Laminin is an important promoter of cell-matrix interactions. A number of active laminin domains have been defined by use of synthetic peptides. The Tyr-Ile-Gly Ser-Arg (YIGSR) sequence on the B1 chain in laminin can decrease tumor growth and metastasis, whereas another sequence containing Ser-Ile-Lys-Val-Ala-Val (SIKVAV) on the A chain can increase tumor growth and metastasis. Here, we selected B16 F10 melanoma cells by adherence or nonadherence to either YIGSR- or SIKVAV-coated dishes and established 3 B16-F10 variants: YIGSR-adherent cells (Y+), YIGSR nonadherent cells (Y-), and SIKVAV-nonadherent cells (S-). SIKVAV-adherent cells were not selected because most of F10 cells attached to the SIKVAV-coated dish. These cell lines proliferated at the same rate as the parent F10 cells and attached equally to laminin, collagen IV, and fibronectin. Y+ cells produced rapidly growing tumors after s.c. injection and twice as many lung colonies as the parental F10 cells after i.v. injection. In contrast, Y- cells produced more slowly growing tumors after s.c. injection and produced one-third of the lung colonies relative to the parent cells after i.v. injection. S- cells produced slowly growing tumors after s.c. injection and yielded similar numbers but smaller colonies in the lung than the parental B16-F10 cells after i.v. injection. These data suggest that interactions of melanoma cells with the YIGSR site on laminin are probably important for both colony formation in a target organ (lung) and subsequent tumor growth, while the SIKVAV-containing site on laminin may be more important for tumor growth. PMID- 8417835 TI - Fifth International Workshop on Ataxia-Telangiectasia. PMID- 8417836 TI - 20-Hydroxyeicosatetraenoic acid is an endogenous vasoconstrictor of canine renal arcuate arteries. AB - Recent studies have indicated that renal arteries can produce 20 hydroxyeicosatetraenoic acid (20-HETE) and suggest the potential involvement of a P450 metabolite of arachidonic acid in the myogenic activation of canine renal arteries. In the present study, the effects of 20-HETE on isolated canine renal arcuate arteries were studied. Administration of 20-HETE to the bath or the lumen at concentrations of 0.01-1 microM produced a graded reduction in the diameter of these vessels. In contrast, 19(R)-HETE was a vasodilator, whereas 19(S)-HETE was relatively inactive. The vasoconstrictor response to 20-HETE was not altered by the cyclooxygenase inhibitor indomethacin, endoperoxide/thromboxane receptor antagonist SQ29548, or combined blockade of the cyclooxygenase, lipoxygenase, and P450 pathways using indomethacin, baicalein, and 7-ethoxyresorufin. The response to 20-HETE was associated with depolarization and a sustained increase in the intracellular calcium concentration in renal vascular smooth muscle cells. Patch clamp studies indicated that 20-HETE significantly reduced mean open time, the open-state probability, and the frequency of opening of a 117-pS K+ channel recorded from renal vascular smooth muscle cells in the cell-attached mode. Microsomes prepared from the renal cortex of dogs produced 20-HETE and 20 carboxyarachidonic acid when incubated with [14C]arachidonic acid. These results indicate that 20-HETE is an endogenous constrictor of canine renal arcuate arteries. The vasoconstrictor response to 20-HETE resembles the myogenic activation of these vessels after elevations in transmural pressure and suggests a potential role for this substance in the regulation of renal vascular tone. PMID- 8417837 TI - Epicardial mapping of ventricular defibrillation with monophasic and biphasic shocks in dogs. AB - To study the mechanism of defibrillation and the reason for the increased defibrillation efficacy of biphasic waveforms, the potential gradient in a 32 x 30-mm region of the right ventricle in 15 dogs was progressively lowered in four steps while a strong potential gradient field was maintained throughout the rest of the ventricular myocardium. The volume of right ventricle beneath the plaque was 10 +/- 2% of the total ventricular mass. A 10-msec monophasic (eight dogs) or 5/5-msec biphasic (seven dogs) truncated exponential shock 30% above the defibrillation threshold voltage was given via electrodes on the left ventricular apex and right atrium to create the strong potential gradient field. Simultaneously, a weaker shock with the same waveform but opposite polarity was given via mesh electrodes on either side of the small right ventricular region to cancel part of the potential difference in the region and to create one of the four levels of potential gradient fields. Shock potentials and activations were recorded from 117 epicardial electrodes in the small region, and in one dog global epicardial activations and potentials were recorded from a sock containing 72 electrodes. Each gradient field was tested 10 times for successful defibrillation after 10 seconds of electrically induced fibrillation. For both monophasic and biphasic shocks, the percentage of successful defibrillation attempts decreased (p < 0.05) as the potential gradient decreased in the small region. Defibrillation was successful approximately 80% of the time for a mean +/ SD potential gradient of 5.4 +/- 0.8 V/cm for monophasic shocks and 2.7 +/- 0.3 V/cm for biphasic shocks (p < 0.05). No postshock activation fronts arose from the small region for eight waveform when the gradient was more than 5 V/cm. For both waveforms, the postshock activation fronts after the shocks were markedly different from those just before the shock and exhibited either a focal origin or unidirectional conduction.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8417838 TI - Effects of lysolipids and oxidatively modified low density lipoprotein on endothelium-dependent relaxation of rabbit aorta. AB - Exposure of isolated arteries to oxidatively modified low density lipoprotein (LDL) has been reported to suppress endothelium-dependent relaxation (EDR). To determine whether lipid degradation products in oxidized LDL contribute to impaired relaxation, we have tested the responsiveness of isolated rabbit aortas to endothelium-dependent relaxants (acetylcholine, ATP, and calcium ionophore A23187) and nitroglycerin before and after 2-hour incubations with selected lipids and LDL preparations. Concentrations (10 microM) of lecithin, phosphatidylserine, lysophosphatidylserine, sphingomyelin, phosphatidic acid, palmitate, arachidonate, and auto-oxidized arachidonate had no effect on EDR. Concentrations (10 microM) of lysolecithin, lyso-platelet activating factor, and sphingosine significantly suppressed endothelium-dependent relaxation. Native LDL (100 micrograms/ml incubation buffer) containing only small amounts of lysophosphatidylcholine exerted no effect on EDR. In contrast, LDL preparations oxidatively modified by exposure to cultured endothelial cells or copper inhibited EDR. When modified LDL was depleted of its lysolecithin by treatment with a selective phospholipase B (lysolecithinase), the inhibitory effects were attenuated. In contrast, native LDL accumulating lysolecithin under the influence of a phospholipase A2 (lecithinase) exerted inhibitory effects mimicking those of oxidized LDL. Lipids and lipoproteins had no effect on the responsiveness to nitroglycerin, an endothelium-independent vasodilator. We conclude that lysolecithin in oxidatively modified LDL contributes importantly to its vasomotor effects. PMID- 8417839 TI - Altered left atrial compliance after atrial appendectomy. Influence on left atrial and ventricular filling. AB - Previous studies have shown regional differences in atrial distensibility. We studied 12 open-chest dogs to test the hypothesis that left atrial compliance is decreased after removal of the left atrial appendage and to determine the effect of altered atrial compliance on atrial reservoir and conduit function. Sonomicrometer crystal pairs were used to measure the long- and short-axis diameters of the left atrium over a wide range of intracardiac pressures and volumes obtained by intravenous hetastarch infusion both before and after suture ligation of the left atrial appendage (appendectomy). Pulmonary venous flow was measured with an ultrasonic flowmeter, and transmitral flow velocities were measured with transesophageal Doppler echocardiography. After appendectomy, the diastolic pressure-volume relation was shifted upward and to the left in six of seven dogs. The mean dynamic stiffness constant of the left atrial diastolic pressure-volume relation was significantly greater after appendectomy than before (0.20 +/- 0.11 [mean +/- SD] versus 0.14 +/- 0.08 ml-1, p < 0.01); the mean y intercept was slightly, but significantly, less after appendectomy (0.6 +/- 0.3 versus 1.3 +/- 0.6 mm Hg, p < 0.05). The left atrial reservoir volume (maximum minus minimum left atrial volume) was significantly less after appendectomy at matched left atrial pressures. The systolic to diastolic flow integral ratio of pulmonary venous flow (JFTI/KFTI), an index of the relative reservoir to conduit functions of the left atrium, increased significantly with volume infusion only before appendectomy; at matched left atrial pressure, JFTI/KFTI was significantly less afterwards.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417840 TI - Substrate-induced changes in the lipid content of ischemic and reperfused myocardium. Its relation to hemodynamic recovery. AB - To investigate the effect of lactate, pyruvate, and glucose on the endogenous levels of lipids in the normoxic, ischemic, and reperfused myocardium, isolated working rat hearts were exposed to various grades of ischemic insult (15, 30, or 45 minutes). Glucose was present as the basal substrate in the perfusion medium, and lactate (5 mM) or pyruvate (5 mM) was added as the cosubstrate. Lipid metabolism was evaluated by fatty acid accumulation, triacylglycerol turnover, and phospholipid homeostasis. Exogenous lactate significantly increased fatty acid content above preischemic levels after 45 minutes of ischemia. In glucose perfused hearts, fatty acid levels were even slightly higher than in lactate perfused hearts, whereas pyruvate-perfused hearts demonstrated less accumulation of fatty acids. By reperfusion, fatty acid levels in glucose-perfused hearts returned to control values. In lactate- and pyruvate-perfused hearts, fatty acid accumulation was further enhanced by reperfusion. When the fatty acid content exceeded 400 nmol/g dry wt during reperfusion, hemodynamic function was impaired, whereas fatty acid levels below 400 nmol/g dry wt did not correlate with hemodynamic recovery. The total triacylglycerol content did not change during ischemia and reperfusion. However, accumulation of glycerol was remarkable during the first 15 minutes of ischemia in all hearts, and release of glycerol by reperfusion was considerable in lactate-perfused hearts after 30 minutes of ischemia and in all groups of hearts after 45 minutes of ischemia. Release of glycerol in association with maintained levels of triacylglycerols suggests turnover of the triacylglycerol pool. The rate of triacylglycerol cycling correlated poorly with hemodynamic recovery. Accumulation of arachidonic acid revealed disturbances in phospholipid turnover. Arachidonic acid accumulation during reperfusion demonstrated a strong relation with impairment of cardiac function. Hence, derangements in phospholipid homeostasis during reperfusion might be involved in myocardial damage, which is influenced by the substrates available. PMID- 8417841 TI - Regional myocardial capillary erythrocyte transit time in the normal resting heart. AB - A major determinant of oxygen transport to the myocardium is the time spent by the erythrocytes (red blood cells [RBCs]) traversing the microcirculation. Although it has been shown that the myocardium has regional differences in blood volume, blood flow, metabolism, and sensitivity to ischemic injury, the regional distribution of RBC transit times through the myocardial capillaries has not been previously measured. The present study was designed to measure the regional myocardial capillary RBC transit time by a new technique to determine whether there are regional differences in the capillary RBC transit time in the normal resting heart. Anesthetized open-chest male New Zealand White rabbits (3.0-3.7 kg, n = 8) were studied. Regional myocardial blood volume was determined using chromium-51-labeled RBCs, and regional blood flow was measured using a reference flow technique and a left atrial injection of 15-microns-radiolabeled (gadolinium 153, 10-20 muCi) microspheres. Capillary blood volume was determined by multiplying the regional blood volume by the histologically determined fraction of the total blood volume that was in the capillaries. Capillary RBC transit time was calculated as the quotient of capillary blood volume and blood flow. The myocardial capillary blood volume was the same in the endocardium and the epicardium (4.67 +/- 0.67 ml/100 g for endocardium versus 4.52 +/- 0.70 ml/100 g for epicardium, p = NS), whereas myocardial blood flow tended to be greater in the endocardium (6.09 +/- 0.73 ml/sec per 100 g for endocardium versus 5.47 +/- 0.75 ml/sec per 100 g for epicardium), although this was not statistically significant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417842 TI - Endothelin as a neuropeptide. Cardiovascular effects in the brainstem of normotensive rats. AB - The relevance of endothelin in central cardiovascular function was studied in urethane-anesthetized Sprague-Dawley rats. Blood pressure (BP) was monitored intra-arterially, and cerebrospinal fluid (CSF) was collected through an intracisternal catheter for radioimmunoassay of endothelin-1 (ET-1). Endothelin levels in the CSF were significantly higher (39 +/- 3 pg/ml) than in plasma (10 +/- 3 pg/ml, n = 11). ET-1 in CSF or plasma was not affected by systemic infusion of saline, but its levels significantly decreased when a sustained increase in BP was elicited with phenylephrine (14 +/- 7 pg/ml in the CSF and 6 +/- 4 pg/ml in plasma, n = 5). In sinoaortic-denervated animals, phenylephrine failed to reduce CSF endothelin levels. In different experiments, intracisternal administration of ET-1 (10 pmol) evoked an initial decrease in BP and heart rate (HR), followed by pronounced hypertension, bradycardia, and, in 70% of the animals, death from cardiorespiratory failure. Intracisternal administration of endothelin-3 (ET-3, 80 pmol, n = 11) evoked only a modest hypotensive and bradycardic response without cardiorespiratory impairment. Microinjection of ET-1 (0.5, 1, 2, 4, and 6 pmol/60 nl) into the nucleus of the solitary tract or area postrema produced a decrease in BP and HR. On the other hand, injection of low concentrations of ET-3 into the nucleus of the solitary tract increased BP and HR (at 2 pmol, 17 +/- 3 mm Hg, 14 +/- 6 beats per minute, n = 7), whereas ET-3 in the area postrema produced a prominent dose-related decrease in BP and HR. In the rostroventrolateral medulla, the lowest doses of ET-1 first modestly increased BP and renal sympathetic nerve activity. These effects were followed by hypotension, bradycardia, increase in respiratory frequency, and further enhancement of sympathetic nerve traffic. In 29% of the animals, these effects were followed by cardiorespiratory arrest. The specificity of the cardiovascular response to endothelin was demonstrated by the inhibitory effects of the receptor antagonist BQ-123. These results demonstrate that endothelin has specific cardiovascular effects in the brainstem of the rat and support a role for endothelin in cardiovascular regulation. PMID- 8417843 TI - Impairment by damage of the pterygopalatine ganglion of nitroxidergic vasodilator nerve function in canine cerebral and retinal arteries. AB - Histochemical study revealed that transcutaneous injection of ethanol into the vicinity of the pterygopalatine ganglion greatly decreased the positive staining for NADPH diaphorase activity after 1 week in the ipsilateral ganglion of a dog and abolished the staining of perivascular nerves in the middle and posterior cerebral arteries. Transmural electrical stimulation or nicotine produced a relaxation in middle and posterior cerebral arteries isolated from the side with the nontreated ganglion (control side), whereas the relaxation was abolished or reversed to a contraction in the arteries from the side with the ethanol-treated ganglion. Nitric oxide-induced relaxations did not differ in the arteries from both sides. The response to nerve stimulation of the control arteries was suppressed by treatment with NG-nitro-L-arginine (L-NA), an inhibitor of nitric oxide synthase, and the inhibition was reversed by L-arginine. Nicotine produced a contraction followed by a relaxation in central retinal arterial strips obtained from the control side; the relaxation was abolished and the contraction was potentiated in the arteries from the treated side. The nicotine-induced relaxation was abolished by L-NA, and the contraction was suppressed by phentolamine. On the other hand, the nicotine-induced relaxation in superficial temporal arteries, susceptible to L-NA, was not attenuated by treatment with ethanol. The findings obtained so far support our hypothesis that nitric oxide released from the vasodilator nerve acts as a transmitter to produce arterial smooth muscle relaxation and suggest that the nerve fibers to the cerebral and retinal arteries arise from the pterygopalatine ganglion. PMID- 8417844 TI - Smooth and skeletal muscle actin are mechanically indistinguishable in the in vitro motility assay. AB - Smooth muscle produces as much stress as skeletal muscle with less myosin. To determine if the actin isoforms specific to smooth muscle contribute to the enhanced force generation, the motility of actin filaments from smooth and skeletal muscle were compared in an in vitro assay in which single fluorescently labeled actin filaments slide over a myosin-coated coverslip. No difference was observed between the velocity of smooth versus skeletal muscle actin filaments over either smooth or skeletal muscle myosin over a large range of assay conditions (changes in pH, ionic strength, and [ATP]). Similarly, no difference was observed between the two actins when the filaments moved under load over mixtures of phosphorylated smooth and skeletal muscle myosin. Thus, it appears that the actin isoforms of smooth and skeletal muscle are mechanically indistinguishable in the motility assay and that smooth muscle's enhanced force generation may originate within the myosin molecule specific to smooth muscle. PMID- 8417845 TI - Induction of vascular smooth muscle cell expression of plasminogen activator inhibitor-1 by thrombin. AB - Local accumulation of plasminogen activator inhibitor-1 (PAI-1) in response to thrombosis has been implicated not only in inhibition of fibrinolysis but also in the pathogenesis of vascular disease. To determine whether thrombin, known to be released from thrombi, can induce expression of PAI-1 in vascular smooth muscle, bovine aortic smooth muscle cells were exposed to highly purified bovine thrombin. Thrombin, in the absence of serum, induced production of PAI-1 by bovine aortic smooth muscle cells in a dose-dependent manner. PAI-1 activity in the conditioned media reached a maximum with 12 nM thrombin. Metabolic labeling with [35S]methionine demonstrated that the elaborated PAI-1 was newly synthesized and that it comprised both a cleaved inactive 42-kd form and an uncleaved active 46-kd form. The increase of PAI-1 activity in the media paralleled the thrombin induced increase in the concentration of the 46-kd form. Preincubation of thrombin with hirudin, a specific inhibitor of thrombin, or with D-phenylalanyl-L prolyl-L-arginine chloromethyl ketone, an inhibitor of the active site of thrombin, prevented the induction of PAI-1 synthesis. The stimulatory effect of thrombin on PAI-1 synthesis was also evident at the level of expression of mRNA, with steady-state PAI-1 mRNA levels increasing by 100% in 4-8 hours. When the bovine aortic smooth muscle cells were exposed to transforming growth factor-beta 1, an agonist shown previously to increase PAI-1 synthesis in diverse cell types, synergy with thrombin was evident.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417846 TI - Altered calcium sensitivity of isometric tension in myocyte-sized preparations of porcine postischemic stunned myocardium. AB - Postischemic ventricular myocardial dysfunction, termed stunning, is characterized by a persistent but ultimately reversible depression of contractile function. The present study was undertaken to investigate the possibilities that reduced contractile force in stunning is due to a decrease in maximal tension generating capability or to a decrease in the Ca2+ sensitivity of the myofilaments. The experiments combine an in vivo open-chest porcine heart model of stunning (n = 5) with in vitro measures of myocyte myofilament calcium sensitivity from these same hearts. Regional myocardial function in the left anterior descending coronary artery (LAD) perfusion bed of porcine hearts was measured with transmural ultrasonic crystals. The protocol was 45 minutes of low flow LAD ischemia at 40% of control flow, followed by 30 minutes of postischemic reperfusion at control aerobic flow. Percent systolic wall thickening decreased to 8 +/- 5% of control during ischemia (p < 0.05) and returned to 38 +/- 8% of control in the postischemic stunned state (p < 0.05). Serial endocardial biopsies were obtained from the preischemic and postischemic myocardium in the LAD perfusion bed and from the aerobically perfused myocardium in the circumflex bed. The biopsies were mechanically disrupted, and myocyte-sized preparations of permeabilized myocardium were attached to a force transducer and a length changing device to allow for direct measurement of steady-state tension-pCa (i.e., -log[Ca2+]) relations. The pCa for half-maximal activation of tension, i.e., pCa50, in LAD myocardium decreased from 5.88 +/- 0.05 before ischemia to 5.69 +/- 0.03 after ischemia (p < 0.05); however, maximal Ca(2+)-activated tension and the slope of the tension-pCa relation were unaffected by the ischemic episode.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417847 TI - Mechanical properties of rat cardiac skinned fibers are altered by chronic growth hormone hypersecretion. AB - Chronic growth hormone (GH) hypersecretion in rats leads to increased isometric force without affecting the unloaded shortening velocity of isolated cardiac papillary muscles, despite a marked isomyosin shift toward V3. To determine if alterations occurred at the level of the contractile proteins in rats bearing a GH-secreting tumor (GH rats), we examined the mechanical properties of skinned fibers to eliminate the early steps of the excitation-contraction coupling mechanism. We found that maximal active tension and stiffness at saturating calcium concentrations (pCa 4.5) were markedly higher in GH rats than in control rats (tension, 52.9 +/- 5.2 versus 38.1 +/- 4.6 mN.mm-2, p < 0.05; stiffness, 1,105 +/- 120 versus 685 +/- 88 mN.mm-2.microns-1, p < 0.01), whereas values at low calcium concentrations (pCa 9) were unchanged. In addition, the calcium sensitivity of the contractile proteins was slightly but significantly higher in GH rats than in control rats (delta pCa 0.04, p < 0.001). The crossbridge cycling rate, reflected by the response to quick length changes, was lower in GH rats than in control rats (62.0 +/- 2.6 versus 77.4 +/- 6.6 sec-1, p < 0.05), in good agreement with a decrease in the proportion of alpha-myosin heavy chains in the corresponding papillary muscles (45.5 +/- 2.0% versus 94.6 +/- 2.4%, p < 0.001). The changes in myosin heavy chain protein phenotype were paralleled by similar changes of the corresponding mRNAs, indicating that the latter occurred mainly at a pretranslational level. These results demonstrate that during chronic GH hypersecretion in rats, alterations at the myofibrillar level contribute to the increase in myocardial contractility observed in intact muscle. PMID- 8417848 TI - Rate-dependent prolongation of cardiac action potentials by a methanesulfonanilide class III antiarrhythmic agent. Specific block of rapidly activating delayed rectifier K+ current by dofetilide. AB - Class III antiarrhythmic agents act by selective prolongation of cardiac action potential duration (APD). Methanesulfonanilide class III agents (e.g., E-4031 and dofetilide) are extremely potent and lengthen action potentials in a "reverse" rate-dependent manner; i.e., effects are greater at low compared with high rates of stimulation. By using the whole-cell current-clamp technique in isolated guinea pig ventricular myocytes, APD was shortened by rapid pacing (244 +/- 16 msec at 30 pulses per minute, 166 +/- 8 msec at 240 pulses per minute; n = 8). Dofetilide (1 microM) prolonged APD more when cells were stimulated at the rate of 30 pulses per minute (44 +/- 10-msec increase) than at 240 pulses per minute (21 +/- 5-msec increase). We investigated the mechanism of APD prolongation using voltage-clamp techniques. Dofetilide selectively inhibited IKr (IC50, 31.5 nM), defined as the rapidly activating inward rectifying component of net delayed rectifier K+ current (IK), without effects on the larger but more slowly activating component of IK (IKs) or on the inward rectifier K+ current (IK1). To examine the rate-dependent effects of dofetilide on APD, trains of conditioning pulses to 0 mV (200-msec duration) were applied at either 30 or 240 pulses per minute to mimic the action potential experiments. Test pulses or ramps were given after the conditioning train to quantitate changes in IK1, IKr, or IKs. The magnitude of neither IK1 nor IKr was dependent on the rate of the preceding train of depolarizations. Sensitivity to block of IKr by dofetilide was rate independent.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8417849 TI - XeCl laser-induced fluorescence of atherosclerotic arteries. Spectral similarities between lipid-rich lesions and peroxidized lipoproteins. AB - Autofluorescence spectroscopy of arterial surfaces provides information about the distribution and composition of atherosclerotic plaques. The aim of the study was to determine whether accumulation of peroxidized lipoproteins in arterial walls, a process postulated to play a role in initiating atherosclerotic changes, can be demonstrated by fluorescence spectroscopy. XeCl excimer laser (308 nm)-induced fluorescence of human aortas containing early lipid-rich noncollagenous lesions exhibited marked red shifts and broadening of the fluorescence spectra compared with spectra from nonatherosclerotic aortas. Similar profiles were observed in spectra obtained from oxidatively modified low density lipoprotein but not native low density lipoprotein. In hypercholesterolemic rabbits with early foam cell lesions, spectral shifts resembled those of oxidized beta-very low density lipoprotein, the major lipoprotein accumulating in arteries of rabbits fed cholesterol. XeCl laser-fluorescence spectroscopy of arterial surfaces may be useful for the identification of arteries accumulating modified lipoproteins (oxidized low density lipoprotein), a chemical change indicative of atherosclerosis in its early and probably reversible stages. PMID- 8417850 TI - Hyperthermia and birth defects. PMID- 8417851 TI - Physiologic differences between twin and single born beef calves in the first two days of life. AB - Behavioral observations and hematological, serum biochemical and blood gas measurements were made on 8 naturally occurring twin calves during the first 48 hours of life. These values were compared to similar measurements collected from 30 single born calves, born under the same calving conditions. All calves survived to at least 3 weeks of age without physically detectable disease. Although the gestational age of the twins and singles were not different, the twins had a lower mean birth weight. Calving difficulty score, time interval to standing and time interval to nursing were not different between the 2 groups. Twin calves had significantly lower rectal temperatures, arterial oxygen tensions and blood glucose concentrations than the single calves through the first 12 hours of life. Hct, Hgb concentration, and RBC were lower in twin calves throughout the 48 hour period. The N:L ratio was lower in the twins at birth. Mean serum IgG1 concentrations were lower in twins only at 24 hours whereas IgM concentrations were lower at both 24 and 48 hours in twins. Serum chemistry value differences between twin and single calves were most numerous at 24 hours of age when serum proteins, urea nitrogen, total bilirubin, sodium, chloride, and total calcium concentrations were higher in the twins and serum phosphorus concentration was lower in the twins. PMID- 8417852 TI - Penetrating lingual foreign bodies in three horses. AB - Three horses examined for complaints of ptyalism and dysphagia were found to have metallic lingual foreign bodies. Metallic foreign bodies were located by oral examination combined with radiography. In 1 horse clinical signs resolved without removal of the foreign body. The foreign body was extracted via an oral approach in the second horse; a mandibular symphysiotomy and radiographic guidance were necessary for removal in the third horse. In all 3 cases, the presenting clinical signs subsided. PMID- 8417853 TI - Acute renal failure in the llama (Lama glama). AB - This clinical report describes 2 cases of acute renal failure in the llama (Lama glama). Both llamas presented with histories of administration of potentially nephrotoxic agents prior to hospitalization. It is suggested that renal function be carefully monitored in all llamas treated with nephrotoxic agents, especially when history or clinical signs indicate increased risk for renal failure. PMID- 8417854 TI - Surgical management of urolithiasis in small ruminants. AB - Twenty-one cases of urolithiasis in small ruminants were examined to evaluate the efficacy of various surgical treatments. Of 21 cases, all but 1 had multiple calculi. Urethral process amputation and medical therapy either did not relieve the obstruction or provided only temporary relief (< 36 hours) in 14 of 16 cases. In 2 cases, obstruction recurred in 1 year and 4 years, respectively, after urethral process amputation. Ten of 13 animals that had perineal urethrostomy (PU) experienced short-term complications, including postoperative hemorrhage, dehiscence of the surgical wound, and subcutaneous accumulation of urine. Urethral stricture occurred in 7 of 9 animals in which follow-up information was obtained, and long-term survival was 55%. Cystotomy alone was performed on 8 of 21 cases, and 7 of 8 (88%) were healthy at the time of follow-up. In order to relieve the obstruction with cystotomy alone, multiple attempts at normograde and retrograde flushing were required. Cystotomy allowed removal of multiple calculi, bidirectional flushing of the urethra, and posed less risk for short-term postoperative complications and urethral stricture. These data suggest that cystotomy was more effective than PU for the management of urolithiasis in small ruminants. PMID- 8417855 TI - Infiltrative lipoma in a quarter horse. AB - A case of infiltrative lipoma in the left flank of a yearling Quarter Horse is described. Rectal palpation, percutaneous ultrasonographic examination, the marbled appearance and poor delineation from surrounding tissues strongly suggested an infiltrative lipoma. This was confirmed by histological examination. Infiltrative lipomas should be included in the differential diagnosis of soft masses located in equine muscle. PMID- 8417856 TI - Oropharyngeal abscessation in two cows secondary to administration of an oral calcium preparation. AB - This paper describes two cases of infection of the soft palate and pharyngeal wall in cattle. The infections were secondary to trauma and laceration received during treatment with an oral calcium preparation which was given to treat post parturient hypocalcemia. Clinical signs included dehydration, depression, pseudoptyalism, and fetid breath. Other signs involved the upper respiratory and gastrointestinal tracts. Digital oral examination revealed the traumatized regions. Treatment included antibiotics and non-steroidal antiinflammatory agents. A semipermanent rumenostomy was performed on one cow for fluid and nutritional maintenance, but she died from secondary septic peritonitis. The second cow survived. PMID- 8417857 TI - Metastatic renal adenocarcinoma in a mule. AB - A 15-year-old mare mule was presented with a 2-month history of weight loss, lethargy, and anorexia. Clinicopathologic abnormalities were consistent with liver disease. Repeated urinalysis revealed hyposthenuria, but water intake and a water deprivation test were normal, suggesting adequate renal function. Ultrasonography revealed pulmonary, hepatic, and renal masses. Cytologic and histologic examination of pulmonary aspirates and hepatic biopsies, respectively, revealed polygonal neoplastic cells with many mitotic figures, the cells having the characteristics of adenocarcinoma. Necropsy confirmed a primary renal adenocarcinoma with metastasis to liver, lung, lymph nodes and the opposite kidney. PMID- 8417858 TI - Abdominal pain associated with an umbilical abscess in a llama. AB - A 3-month-old llama with a presenting complaint of lethargy, anorexia, and a painful, distended abdomen was evaluated. The llama had intermittently strained to defecate during the 3 days prior to admission. Physical examination results, hematologic data and lateral abdominal radiographs were used to diagnose a large umbilical abscess, which was causing a partial obstruction of the gastrointestinal tract. Under general anesthesia, 3 liters of purulent exudate were drained from the abscess. The abscess cavity was then lavaged with saline solution and its capsule was marsupialized to the skin. Cultures of the abscess content yielded Proteus sp, Streptococcus equisimilis, and Clostridium septicum. Two days after surgery, the llama was drinking, eating, and passing feces. The abscess was lavaged daily for a total of 11 days. Six months after surgery, the llama was the same size as other llamas of the same age, and the owners were pleased with the cosmetic appearance of the ventral abdomen. Umbilical abscesses can vary in size and clinical presentation; they should be recognized as a possible cause of abdominal pain with a potential for causing intestinal obstruction in llamas. PMID- 8417859 TI - Squamous cell carcinoma of the anal and perianal area in a bull. AB - A squamous cell carcinoma located adjacent to the anus was diagnosed in a 15-year old light colored Longhorn bull. The tumor restricted the anal orifice to a diameter of 3 cm. Upon histological evaluation, islands of squamous cells were present deep in the dermis and the submucosal connective tissue. It was not possible to determine whether the tumor originated from the perianal region or the anus. This is the first diagnosed and reported occurrence in North America of squamous cell carcinoma in the anal region of a bull. PMID- 8417860 TI - Intracorporeal respiratory support. A potential supplement to airway pressure therapy? PMID- 8417861 TI - Colchicine in the treatment of pulmonary fibrosis. AB - There is no standard or known optimal treatment for idiopathic pulmonary fibrosis. Corticosteroids have been used with widely variable benefit. Colchicine has been reported to suppress fibroblast growth factors and to inhibit collagen deposition. A potential role has been proposed for colchicine in the treatment of fibrotic pulmonary diseases. We retrospectively assessed the outcome of 23 patients with idiopathic pulmonary fibrosis in whom colchicine was used as empiric treatment. Eighteen patients had received prior corticosteroid therapy. By clinical and pulmonary function criteria, five patients (22 percent) improved following colchicine, nine (39 percent) remained stable, and nine (39 percent) worsened. The average duration of follow-up was 22 months. Despite limitations of this retrospective analysis, colchicine may be of benefit in pulmonary fibrosis and should be considered for further clinical trials. PMID- 8417862 TI - Pulmonary function and hypoxic ventilatory response in subjects susceptible to high-altitude pulmonary edema. AB - To determine if spirometric changes reflect early high-altitude pulmonary edema (HAPE) formation, we measured the FVC, FEV1, and FEF25-75 serially during the short-term period following simulated altitude exposure (4,400 m) in eight male subjects, four with a history of HAPE and four control subjects who had never experienced HAPE. Three of the four HAPE-susceptible subjects developed acute mountain sickness (AMS), based on their positive Environmental Symptom Questionnaire (AMS-C) scores. Clinical signs and symptoms of mild pulmonary edema developed in two of the three subjects with AMS after 4 h of exposure, which prompted their removal from the chamber. Their spirometry showed small decreases in FVC and greater decreases in FEV1 and FEF25-75 after arrival at high altitude in the presence of rales or wheezing on clinical examination and normal chest radiographs. One of the two subjects had desaturation (59 percent) and tachycardia during mild exercise, and excessive fatigue and inability to complete the exercise protocol developed in the other at 4 h. The six other subjects had minimal changes in spirometry and did not develop signs of lung edema. Further, we measured each subject's ventilatory response to hypoxia (HVR) prior to decompression to determine whether the HVR would predict the development of altitude illness in susceptible subjects. In contrast to anticipated results, high ventilatory responses to acute hypoxia, supported by increased ventilation during exposure to high altitude, occurred in the two subjects in whom symptoms of HAPE developed. The results confirm that HAPE can occur in susceptible individuals despite the presence of a normal or high ventilatory response to hypoxia. PMID- 8417863 TI - Comparison of blue dye visualization and glucose oxidase test strip methods for detecting pulmonary aspiration of enteral feedings in intubated adults. AB - STUDY OBJECTIVE: To compare the relative utility of blue dye visualization with a glucose oxidase test strip method for detecting aspiration of enteral feedings. DESIGN: Tracheally intubated adults were prospectively monitored for aspiration of enteral feedings. SETTING: Intensive care units of two community hospitals in Michigan. INTERVENTIONS: None. PATIENTS: The experimental group consisted of 15 patients receiving enteral feedings. The control group included 14 patients not enterally fed. MEASUREMENTS AND RESULTS: Blue food coloring was added to feeding formulas to obtain a visible blue color. At 8-h intervals, tracheal secretions were examined for blue discoloration, followed by measurement of glucose concentration using a calibrated glucose meter. Clinically significant aspiration was defined to require the following: (1) a bloodless positive glucose reading (> or = 20 mg/dl); (2) one or more signs of systemic inflammation; and (3) one or more signs of respiratory deterioration. Eight (53 percent) of 15 patients in the experimental group experienced at least one episode of presumptive aspiration as defined by either a bloodless positive glucose reading or visible blue discoloration of tracheal secretions. Clinically significant aspiration occurred in 5 (33 percent) of 15 patients in whom bloodless glucose readings were positive in 13 (19 percent) of 67 samples; among patients not developing this complication, glucose was found in only 3 (5 percent) of 60 samples; (p = 0.005). Inspecting tracheal secretions for blue dye usually failed to detect aspiration episodes identifiable by the glucose oxidase test strip method (relative sensitivity, 13 percent). Blue dye visualization performed no better among patients developing clinically significant aspiration (relative sensitivity, 15 percent). Patients who developed clinically significant aspiration received more of their enteral feedings in the supine position than patients without this complication (98 percent vs 21 percent; p < 0.001). CONCLUSIONS: Inspecting tracheal secretions for blue discoloration failed to detect most episodes of enteral feeding aspiration. Glucose oxidase test strip methods should replace blue dye visualization for detecting aspiration of enteral feedings in intubated adults. PMID- 8417864 TI - Pulmonary arterial hypertension and cor pulmonale associated with chronic domestic woodsmoke inhalation. AB - We describe the clinical, radiologic, functional, and pulmonary hemodynamic characteristics of a group of 30 nonsmoking patients with a lung disease that may be related to intense, long-standing indoor wood-smoke exposure. The endoscopic and some of the pathologic findings are also presented. Intense and prolonged wood-smoke inhalation may produce a chronic pulmonary disease that is similar in many aspects to other forms of inorganic dust-exposure interstitial lung disease. It affects mostly country women in their 60s, and severe dyspnea and cough are the outstanding complaints. The chest roentgenograms show a diffuse, bilateral, reticulonodular pattern, combined with normalized or hyperinflated lungs, as well as indirect signs of pulmonary arterial hypertension (PAH). On the pulmonary function test the patients show a mixed restrictive-obstructive pattern with severe hypoxemia and variable degrees of hypercapnia. Endoscopic findings are those of acute and chronic bronchitis and intense anthracotic staining of the airways appears to be quite characteristic. Fibrous and inflammatory focal thickening of the alveolar septa as well as diffuse parenchymal anthracotic deposits are the most prominent pathologic findings, although inflammatory changes of the bronchial epithelium are also present. The patients had severe PAH in which, as in other chronic lung diseases, chronic alveolar hypoxia may play the main pathogenetic role. However, PAH in wood-smoke inhalation-associated lung disease (WSIALD) appears to be more severe than in other forms of interstitial lung disease and tobacco-related COPD. The patients we studied are a selected group and they may represent one end of the spectrum of the WSIALD. PMID- 8417865 TI - Preventive effects of beclomethasone on histamine-induced changes in breathing pattern in asthma. AB - Bronchial mucosa inflammation is a hallmark of asthma. Epithelial damage due to inflammatory process may contribute to induce a pattern of rapid and shallow breathing (RSB). Probably due to its effects on inflammatory process, beclomethasone dipropionate (BDP) decreases bronchial hypersensitivity (BH), as assessed in terms of histamine concentration causing a 20 percent FEV1 decrease from saline solution (PC20FEV1); however, no data are available on the effect of BDP on RSB. We studied 32 asymptomatic asthmatic subjects with a severe to moderate levels of BH (PC20FEV1 0.01 to 1.7 mg/ml). After they were randomly assigned to one month of either BDP (2 mg daily, 17 patients) or placebo (15 patients), they inhaled progressively doubling concentrations of histamine phosphate by tidal breathing method. With histamine in seven BDP-treated and in five placebo-treated patients, decrease in FEV1 > or = 20 percent from saline solution was paralleled by a significant decrease in tidal volume (VT), inspiratory time (Ti), and expiratory time (Te), and increase in respiratory frequency (RF). In the remaining patients, histamine failed to change the breathing pattern. In the seven RSB patients, BDP resulted in a smaller VT decrease (p < 0.02) and a smaller RF increase (p < 0.02) with histamine. The five RSB placebo-treated patients were then given one month BDP (2 mg daily): inhaled BDP, but not placebo, resulted both in a significant increase in PC20FEV1 and modulation in histamine-induced changes in breathing pattern. We conclude that high doses of BDP seem to be able to modulate histamine-induced RSB, an effect that might be linked to reversal of airway inflammation. PMID- 8417866 TI - End-tidal CO2 analysis in sleep apnea syndrome. Conditions for use. AB - The diagnosis of sleep apnea syndrome (SAS) requires expensive and complex instrumentation. The purpose of the present study was to determine the value of end-tidal CO2 (EtCO2) in screening for sleep apneas. Thirty-nine patients referred to our sleep laboratory because of suspected SAS and ten normal subjects were studied. The EtCO2 was measured using an infrared spectrometer (POET) designed for simultaneous measurement of CO2 and pulse oximetry. In 29 subjects, expired gas was sampled with a nasobuccal mask (Respiron) with lateral orifices. In the other 20 subjects, sampling was done with nasobuccal prongs (Criticare) comprising a four-channel plastic tube to the mouth and the nostrils. Data from an 8-h night were transferred the following day to a microcomputer (Apple Macintosh) for processing. Apnea was defined as an absence of detection of CO2 for more than 10 s. Conventional polysomnography was performed (Respisomnographe). The number of apneas in 8 h and the apnea index (number of apneas in 1 h) were calculated after visual analysis on the screen of the polysomnograph and also with EtCO2 analysis. For recordings made with a nasobuccal mask, the regression curve between the apnea indices computed with EtCO2 and polysomnography was an order 2 polynomial curve (r = 0.76; p < 0.001), with an inflection point at 39 apneas per hour. For recordings with nasobuccal prongs, the correlation was very significant (r = 0.95; p < 0.0001), and the regression curve was linear. The EtCO2 with nasobuccal prongs appears to be a simple and reliable method for screening for SAS. PMID- 8417867 TI - Infection of pulmonary artery catheters. Epidemiologic characteristics and multivariate analysis of risk factors. AB - Sixty-nine consecutive pulmonary artery catheters (PACs) were prospectively studied in a medical-surgical intensive care unit. Fifteen (21.7 percent) and two (2.9 percent) of the PACs were associated with colonization and bacteremia, respectively. These data represent an incidence of 4.98 and 0.66 episodes per 100 catheterization-days, respectively. Coagulase-negative staphylococci were the most common isolates. The source of the colonizing microorganism was the skin in 56 percent of cases, hubs in 28 percent, and unknown in 16 percent. From multivariate analysis, only more than 5 days of catheterization was significantly associated with a greater risk of colonization. Antimicrobial use was associated with negative cultures. The most useful method to diagnose colonization was the examination of both tip and intradermal segments. In addition, we suggest associate hub cultures when catheter-related bacteremia is suspected. These data may be useful in improving efficacy in the diagnosis and prevention of PAC related infection. PMID- 8417868 TI - A limited axillary thoracotomy as primary treatment for recurrent spontaneous pneumothorax. AB - Recurrent spontaneous pneumothorax often requires surgical treatment following variable periods of chest tube therapy. A limited axillary thoracotomy provides sufficient exposure to isolate or excise pulmonary blebs and perform a pleurodesis. Prompt use of this surgical approach in lieu of the initial placement of a thoracostomy tube avoids prolonged hospitalization and a significant failure rate of thoracostomy tubes to resolve this problem. This operation can also prevent recurrence, a significant problem for this pathologic process. Fourteen patients with recurrent spontaneous pneumothorax underwent an axillary thoracotomy as either primary treatment or within 72 h of thoracostomy tube placement. The average follow-up was 38 months for the initial 10 patients and 23 months for the entire group. The procedure averaged 66 min in duration. The average incision was 3.3 cm in length. There was an equal male/female ratio and right-left distribution. The patients were discharged an average of 4.2 days after surgery. There were no complications. The most recent six patients with a recurrent pneumothorax were surgically treated on the day of admission without a preoperative chest tube. The other eight patients had a thoracostomy tube for control of the pneumothorax, with surgery performed within 72 h of tube placement. A limited axillary thoracotomy corrected the underlying pathology, hastened hospital discharge, limited pain, prevented short-term recurrence, and was cosmetically acceptable. A limited axillary thoracotomy is the operation of choice when a spontaneous pneumothorax requires surgery. This surgical approach has become our primary treatment for recurrent pneumothorax, avoiding the use of a preoperative thoracostomy tube and unnecessary delay, with excellent results for the patient. PMID- 8417869 TI - Effect of nasal pressure support ventilation and external PEEP on diaphragmatic activity in patients with severe stable COPD. AB - Nasal pressure support ventilation (NPSV) has been shown to be useful in the treatment of acute and chronic pulmonary failure. However, little is known about respiratory muscle activity during NPSV in stable patients with COPD. The aim of this study was to test the effect of two levels (10 and 20 cm H2O) of NPSV on diaphragmatic activity, in a group of seven stable, severe COPD patients (FEV1 20 percent +/- 7 of pred, FEV1/FVC 35 percent) with hypercapnic respiratory insufficiency. Since these patients had an intrinsic PEEP (PEEPi) of 2.6 +/- 1.3 cm H2O, we also investigated the effect of adding 5 cm H2O external PEEP (PEEPe) during NPSV. Blood gases, ventilatory pattern by inductive plethysmography, integrated electromyogram of the diaphragm (Edi), transdiaphragmatic pressure (Pdi), and the diaphragmatic pressure time product (PTPdi) were recorded during randomized 15-min runs of both levels of NPSV with and without the addition of PEEPe. Minute ventilation did not change with the application of NPSV, but a significant decrease in breathing frequency with a parallel increase in tidal volume was observed, so that blood gas determinations improved at the higher levels of support. A marked statistically significant reduction in diaphragmatic activity, as assessed by a decrease in Pdi swings, PTPdi, and Edi, was detected at the levels of 10 and 20 cm H2O; a further significant decrease in these values was observed when PEEPe was added. PEEPi decreased significantly only with the application of PEEPe, resulting in a small increase in end-expiratory lung volume. We conclude that NPSV improves diaphragmatic function in patients with severe stable COPD; this effect may be enhanced by the applications of external PEEP. PMID- 8417870 TI - Trends in physician-diagnosed asthma prevalence in Manitoba between 1980 and 1990. AB - We attempted to assess recent changes in the prevalence of physician-diagnosed asthma and the possible influence of diagnostic exchange on these trends. The routinely collected data of the provincial Health Insurance Plan (physicians' claims) were used to determine the annual prevalence of physician-diagnosed asthma in Manitoba. Results indicate that the prevalence of physician-diagnosed asthma increased for all age groups in both male and female subjects between 1980 and 1990. The average increases were the highest in the age group 5 to 14 years for both sexes. The average increases varied with age and there were significant differences between the two sexes. There was evidence of increasing diagnostic exchange, that is, a tendency to label patients with asthma instead of alternative diagnoses. This was particularly prominent in those younger than 35 years of age. However, the increased prevalence of physician-diagnosed asthma, even for the younger population, cannot be fully explained by diagnostic exchange. PMID- 8417871 TI - Prolonged intracorporeal support of gas exchange with an intravenacaval oxygenator. AB - Patients with severe respiratory failure undergoing conventional mechanical ventilation typically are exposed to levels of ventilator support that place the patient at risk of barotrauma or oxygen toxicity. In severe cases, gas exchange may be inadequate despite maximal ventilator support. We report two cases of advanced respiratory failure in which augmentation of gas exchange was provided for prolonged periods (18 and 13 days) with an intravenacaval hollow-fiber membrane oxygenator (IVOX). Following implantation, significant reductions in ventilator support were possible with improvement in arterial blood gas values. No significant complications were noted. IVOX can provide clinically useful augmentation of gas exchange in respiratory failure and can be used for prolonged periods. PMID- 8417872 TI - Functional electrical stimulation to enhance cough in quadriplegia. AB - Respiratory problems are a major cause of death in the acute and chronic phases of cervical spinal cord injury (CSCI); CSCI paralyzes the intercostal and abdominal muscles, reducing ability to cough and clear secretions. Impaired cough due to neuromuscular disorders can be assessed with the maximum expiratory pressure (MEP). This study consists of two experiments with CSCI patients. In the first, MEP measurements were recorded with the following maneuvers performed: (1) spontaneous cough attempts, (2) manually assisted cough, and (3) cough attempts with functional electrical stimulation (FES) applied to the abdominal wall. In the second, spontaneous cough attempts and cough attempts with a portable FES unit were recorded. These CSCI patients were found to have a greatly reduced MEP when they coughed spontaneously. Either FES-assisted or manually assisted coughing increased the MEP in all patients studied. By increasing the MEP, abdominal muscle FES could enhance cough in quadriplegics. PMID- 8417873 TI - Spirometry and maximal respiratory pressures in patients with facial paralysis. AB - The study included 17 patients with facial paralysis (FP) (10 male, 7 female) aged 47.6 +/- 21.0 years. Twelve of the 17 patients had Bell's palsy. The other causes of FP were as follows: one, polyradiculoneuritis; one, war injury; one, cerebrovascular accident; one, sarcoidosis; and one, dermatomyositis. Spirometry and maximal respiratory pressures (PImax and PEmax) were performed with three different techniques: without holding the lips, with patient holding lips, and with technician holding lips. We observed significant differences for PImax and PEmax among the three methods. There were no differences for spirometric values. In nine of the patients with Bell's palsy, PImax, PEmax, and spirometry tests were repeated two months after the first determination. With respect to the first determination, the values of Pmax-w/v holding lips increased, yet spirometric values were similar. In conclusion, PImax and PEmax can be an index of clinical FP evolution. Spirometric maneuvers can be performed with either patient or technician holding lips to ensure a perfect seal between lips and mouthpiece. PMID- 8417874 TI - Intermittent positive pressure ventilation via the mouth as an alternative to tracheostomy for 257 ventilator users. AB - Despite wider application of the use of nocturnal intermittent positive pressure ventilation (IPPV) via nasal access for the management of nocturnal hypoventilation, there continues to be a lack of familiarity with the use of IPPV via the mouth for ventilatory support. Unlike nasal IPPV, which is generally practical only for nocturnal use, up to 24-h mouth IPPV was the key method of noninvasive ventilatory support that permitted the avoidance or elimination of tracheostomy for 257 individuals with acute or chronic ventilatory failure. Mouth IPPV was delivered via commercially available mouthpieces for daytime aid and mouthpiece with lip seal or custom orthodontic interfaces for nocturnal support. The use of mouth IPPV alone or in a regimen with other noninvasive ventilatory aids was reviewed for these 257 individuals. Mouth IPPV was used for nocturnal aid by 163 individuals, 61 of whom had little or no measurable vital capacity or significant ventilator-free breathing time, for more than 1,560 patient-years with few complications. It was also the predominant method of daytime ventilatory support for 228 individuals for more than 2,350 patient-years. We conclude that for individuals with adequate bulbar muscle function but chronic respiratory muscle insufficiency, mouth IPPV can be an effective alternative to tracheostomy. It can significantly prolong survival while optimizing convenience, safety, and communication. PMID- 8417875 TI - Decrease of skin and bronchial sensitization following short-intensive scheduled immunotherapy in mite-allergic asthma. AB - Changes induced by high-dose intensive specific immunotherapy (ISI) were studied in 18 patients with mite-allergic bronchial asthma and compared with 18 control patients. A biologically standardized well-characterized Dermatophagoides pteronyssinus extract of known potency was used both for in vivo tests and ISI. Immediate D pteronyssinus skin tests (measured by parallel-line bioassay) were reduced 20-fold in ISI treated patients, and delayed skin responses significantly decreased and even disappeared following ISI. Bronchial tolerance to D pteronyssinus increased an average of ten times in ISI-treated patients, whereas nonspecific hyperreactivity remained unchanged. ISI induced significant specific IgG and IgG4 increases but no changes were observed in specific IgG1,2 and 3 or in specific IgE. Clinical score did not change significantly after ISI, short ISI schedule with a well-characterized mite extract greatly reduced the degree of mite sensitization (skin and bronchial allergic responses), but clinical disease failed to improve significantly, probably owing to the lack of influence of ISI in nonspecific hyperreactivity. PMID- 8417876 TI - Iodinated glycerol and thyroid dysfunction. Four cases and a review of the literature. AB - Iodinated glycerol (Organidin) has recently been shown to be a useful adjunct in the symptomatic treatment of patients with chronic obstructive pulmonary disease (COPD). Over an 18-month period, we saw four patients with significant thyroid dysfunction resulting from iodinated glycerol use. All were elderly patients with COPD who had been started on standard doses of iodinated glycerol 4 to 24 months earlier. None of the patients had histories of thyroid disease. Three of the patients had symptomatic thyrotoxicosis and one had severe hypothyroidism while taking iodinated glycerol. We review the literature on the mechanisms and management of iodine-induced thyroid dysfunction and conclude the following: (1) all iodine-containing drugs may adversely affect thyroid function; (2) these compounds should be prescribed with extreme caution to any patients with underlying thyroid disease; and (3) all patients receiving iodine-containing medications should be closely monitored for evidence of thyroid dysfunction. PMID- 8417877 TI - Theophylline effect on sleep in normal subjects. AB - In an effort to isolate the effect of theophylline on sleep from the effect of asthma, we examined the impact of oral theophylline on sleep quality in normal, nonasthmatic subjects. Ten healthy, nonsmoking, male subjects ranging in age from 23 to 35 were studied. The subjects were randomly divided into two groups and studied in a double-blinded, crossover designed protocol. Each group underwent two consecutive nights in the sleep laboratory. Group one received four doses of 300 mg of sustained-release theophylline at 12-h intervals prior to the study night. Group two received a placebo. After 14 days, the patients crossed over to the other group, and the procotcol was repeated. The mean serum theophylline level on the morning following the study night was 7.85 +/- SD 2.12 micrograms/ml, with a range of 5.1 to 12.1 microns/ml. The analysis of variance for a crossover design showed no order effect. The analysis for drug effect showed that theophylline administration resulted in a statistically significant adverse effect on arousals per sleep hour (19.3 vs 15.9 for placebo, p = .006) and total sleep time (370.9 min vs 399.45 min for placebo, p = .015). Comparing the sum of ranks data, theophylline was found to have a significant adverse effect on sleep quality (p = .036). We conclude that low doses of oral theophylline result in a significant disturbance in sleep quality in normal nonasthmatic subjects. PMID- 8417878 TI - The burgeoning field of respiratory care captured in a single volume! A historical perspective and comment on a magnificent new book. PMID- 8417879 TI - Primary pulmonary lymphomas. A clinical study of 70 cases in nonimmunocompromised patients. AB - We studied 70 patients with biopsy-proven pulmonary non-Hodgkin's lymphomas without extrathoracic involvement or mediastinal adenopathy to determine the clinical, imaging, and endoscopic features of this condition in a homogeneous series. In low-grade (LG) lymphomas, symptoms were cough, dyspnea, chest pain, hemoptysis. Imaging features consisted of localized alveolar opacities, infiltrative diffuse opacities, atelectasis, and pleural effusions. Inflammatory changes of the mucosa were present in some patients, leading to bronchial stenosis in 7; biopsies showed lymphomatous infiltration in 12. Prognosis of LG lymphomas was excellent, with 93.6 percent survival at five years. High-grade lymphomas differed from LG lymphomas principally by a more aggressive course and a worse survival. Inflammatory changes occurred in seven of nine cases leading to stenosis in two, and biopsies showed lymphomatous involvement in five. The profile of primary pulmonary lymphomas in this study could help clinicians consider this condition and prompt them to evaluate new diagnostic tools. PMID- 8417880 TI - Intrapleural recombinant IL-2 in passive immunotherapy for malignant pleural effusion. AB - The purpose of this phase 1 study was to determine the toxicity and effectiveness of recombinant interleukin-2 (RU 49637 Roussel Uclaf-France) administered by continuous pleural infusion for 5 days to patients with different histologic subtypes of pleural cancer. Incremental doses of rIL-2 from 3 x 10(6) to 24 x 10(6) were given via a thin catheter inserted into the homolateral pleural cavity. Patients were evaluated before treatment and 36 days after treatment by computed tomography scan and thoracoscopy with biopsy. Twenty-two patients with malignant pleural effusion (15 malignant pleural mesotheliomas, 6 adenocarcinomas, 1 squamous cell carcinoma) were treated. The maximum tolerated dose (MTD) of rIL-2, defined as the dose that produced grade 3 or greater toxic reactions in 50 percent of the patients, was 24 x 10(6) IU/m2/d. Although some side effects were encountered at any dose, tolerance was acceptable. The main side effect was fluid retention (8 of 22) which never exceeded 10 percent of body weight. Responses were achieved in 10 out of 22 patients with 1 complete remission (mesothelioma) and 9 partial remissions (3 adenocarcinomas and 6 malignant pleural mesotheliomas). Based on these results, we recommend that phase 2 studies using rIL-2 at a dose of 21 x 10(6) IU/m2/d via the intrapleural route be undertaken. PMID- 8417881 TI - Substance abuse-related admissions to adult intensive care. AB - The frequency of adult surgical and medical intensive care unit (ICU) admissions related to substance abuse was determined at a large community, trauma, and tertiary referral hospital. Of 435 ICU admissions, 14 percent (95 percent confidence interval [CI], 5 to 23 percent) were tobacco related generating 16 percent of costs, 9 percent (95 percent CI, 0 to 18 percent) were alcohol related generating 13 percent of costs, and 5 percent (95 percent CI, 0 to 14 percent) were illicit drug related generating 10 percent of costs. In all, 28 percent (95 percent CI, 20 to 36 percent) of ICU admissions generating 39 percent of costs were substance abuse related. Substance abuse-related admissions were significantly longer and more costly than admissions not related to substance abuse (4.2 days vs 2.8 days, p = 0.004; $9,610 vs $5,890, p = 0.001). Frequency of substance abuse-related admission was linked with the patient's insurance status (Medicare, private insurance, uninsured). In the uninsured group, 44 percent of admissions were substance abuse related (95 percent CI, 35 to 52 percent), significantly higher than in the private insurance and Medicare groups, and generating 61 percent of all ICU costs in the uninsured group. Large fractions of adult ICU admissions and costs are substance abuse related, particularly in uninsured patients. PMID- 8417882 TI - Latissimus dorsi cardiomyoplasty. Perioperative management and postoperative evolution. AB - Twelve men aged 45 to 69 years, NYHA class 3 or 4 with low isotopic ejection fraction (18 +/- 7 percent), underwent cardiomyoplasty. Eight required cardiopulmonary bypass to treat an associated cardiac lesion. Preoperatively, all patients needed inotropic support with dobutamine and half of them vasodilators, increasing cardiac index by nearly 100 percent. The SvO2 remained over 67 percent during the different stages of the surgical procedure. The mean operating time was 438 +/- 75 min. None of the patients required intra-aortic balloon counterpulsation. Inotropic and vasodilator support was continued in the ICU and appeared especially important during weaning from mechanical ventilation. The average stay in ICU was 6.8 +/- 4.0 days. Three patients died of cardiac failure respectively 8, 11 and 15 months after CMP. One patient underwent transplantation. The eight other surviving patients showed clinical improvement from the third month, but objective criteria for hemodynamic improvement were noted only after one year. Cardiomyoplasty can be an alternative treatment for selected cases of cardiomyopathy. PMID- 8417883 TI - High serum albuterol levels and tachycardia in adult asthmatics treated with high dose continuously aerosolized albuterol. AB - To study the feasibility of using high-dose continuously aerosolized albuterol aerosol in adults, seven adult asthmatic patients were treated eight times with 0.4 mg/kg/h albuterol delivered by continuous nebulization over 4 h. One patient withdrew at 3 h after supraventricular tachycardia developed. This subsided promptly on discontinuing albuterol therapy. Heart rate increases were observed in six of eight treatments and serum albuterol levels at the end of treatment were greater than 25.0 ng/ml in all but one treatment. A mean increase in heart rate of 16.3 percent was observed for the entire group. Of the treatments with elevated (> 25.0 ng/ml) serum albuterol levels, a significant cumulative increase in heart rate was observed with time. A significant improvement of FEV1 was observed (p = 0.0025) with a net increase of 36.8 percent. These data suggest that high-dose continuously aerosolized albuterol treatment in some adult asthmatics can result in markedly elevated serum albuterol levels and potential cardiac stimulation despite spirometric improvement. PMID- 8417884 TI - Prevalence and severity of Doppler-detected valvular regurgitation and estimation of right-sided cardiac pressures in patients with normal two-dimensional echocardiograms. AB - To assess the prevalence and severity of Doppler-detected valvular regurgitation, as assessed by multiple Doppler modalities, in patients with structurally normal hearts, we analyzed Doppler echocardiograms in a consecutive sample of 206 referred patients who were found to have completely normal M-mode and two dimensional echocardiograms. Valvular regurgitation was detected by Doppler in 94 percent, and 56 percent had regurgitation in at least two valves (mitral, tricuspid, and/or aortic). Mitral, tricuspid, and aortic regurgitation was detected in 73 percent, 68 percent, and 12 percent, respectively, with moderate regurgitation occurring in 6 percent, 5 percent, and 2 percent, respectively. The presence of mitral and tricuspid regurgitation was not related to age, although the prevalence of moderate regurgitation was three times more prevalent (p < 0.05) in those > 50 years old compared with those < or = 50 years. Aortic regurgitation was two to three times more prevalent (p < 0.01) in patients > 50 years compared with younger subjects, and moderate aortic regurgitation was three times more prevalent in older patients. Of those with measurable right-sided cardiac pressures, estimated right atrial pressure was < 10 mm Hg in 93 percent of patients, and estimated pulmonary artery systolic pressure was < or = 30 mm Hg in 57 percent of patients. Estimated right atrial pressure was > 10 mm Hg in only 7 percent, and only 13 percent had estimated pulmonary artery systolic pressure > or = 40 mm Hg. These data indicate a very high prevalence of trivial and mild mitral and tricuspid regurgitation in patients with otherwise "normal" hearts, suggesting that these findings are physiologically normal. These data should be considered when addressing management in patients with Doppler-detected valvular regurgitation in order to prevent "iatrogenic heart disease." PMID- 8417885 TI - A three-year study of severe community-acquired pneumonia with emphasis on outcome. AB - Fifty-eight consecutive patients with severe community-acquired pneumonia were studied prospectively during a three-year period. The group included 44 men and 14 women (mean age: 45.0 +/- 15.7 years). The cause of pneumonia was diagnosed in 35 (60.3 percent) cases, and the most common pathogens were Streptococcus pneumoniae (37.1 percent), Legionella pneumophila (22.8 percent) and Gram negative bacilli (11.4 percent). The fact that Mycobacterium tuberculosis was present in four (11.4 percent) patients and Pneumocystis carinii in three (8.5 percent) is worthy of note. The overall death rate was 22.4 percent. More than 50 percent of deaths occurred within the first five days and were caused by septic shock, hemoptysis (tuberculosis) or hypoxia. However, hypoxia remains the main fatal complication and all late-occurring deaths (> 5 days) observed were due to this cause. These data could be important in planning strategies and protocols to improve prognosis. PMID- 8417886 TI - A reappraisal of blind bronchial sampling in the microbiologic diagnosis of nosocomial bronchopneumonia. A comparative study in ventilated patients. AB - STUDY OBJECTIVE: To assess the usefulness of fiberscopy for microbiologic diagnosis of nosocomial bronchopneumonia (NBP) in ventilated patients. DESIGN: Data were collected prospectively. We compared the results of semiquantitative cultures obtained by protected specimen brush (PSB), bronchoalveolar lavage (BAL) and blind bronchial sampling (BBS). Positive thresholds were 10(3) CFU ml-1 for PSB and BAL and 10(4) CFU ml-1 for BBS. We also evaluated the diagnostic performance of direct examination of samples obtained by BAL and BBS. PATIENTS: We carried out this study in 64 ventilated patients admitted to a medico-surgical ICU. RESULTS: During the study, 85 sets of samplings were obtained. The concordance between the results of specimen cultures obtained with the three techniques was 87 percent. The concordance between BBS and PSB or between BBS and BAL was 91.8 percent. In two of seven patients with discordant results between BBS and PSB, the microorganisms isolated from blood cultures were found on BBS, but not on PSB samples. As for direct examination, the thresholds for the diagnosis of NBP using BBS were as follows: > or = 10 polymorphonuclear neutrophils (PMN)/high-power field (HPF), > or = 1 bacteria/oil immersion field (OIF), presence of intracellular bacterial inclusions. Using BAL, the thresholds were as follows: > or = 1 PMN/HPF, presence of bacteria/OIF, presence of intracellular bacterial inclusions. The specificity of the presence of bacterial inclusions was excellent regardless of the sampling technique, but the sensitivity of this criteria was mediocre (30.8 percent with BBS and 19.2 percent with BAL). Except for the number of PMN on BBS, all the other diagnostic criteria (PMN count on BAL, bacterial count, count of cells exhibiting inclusions) provide a similar prediction of NBP (correctly classified: 61.2 to 81.2 percent). No combination of criteria enabled significantly better classification regardless of the sampling technique. CONCLUSIONS: In view of these findings and the high cost and morbidity of fiberscopy, it is arguably better to use a simple, repeatable, and risk-free technique for obtaining culture specimens from mechanically ventilated patients. Obviously, protected brushing techniques remain the most effective for nonintubated patients. PMID- 8417887 TI - Protected specimen brush in the assessment of ventilator-associated pneumonia. Selection of a certain lung segment for bronchoscopic sampling is unnecessary. AB - The protected specimen brush (PSB) with quantitative cultures is one of the most reliable techniques for assessing pneumonia in mechanically ventilated (MV) patients. The need to select a certain lung segment for bronchoscopic sampling is still debated. We investigated whether the results of PSB specimens collected within an area radiographically involved with pneumonia (inv-PSB) differed from the results of PSB specimens collected within a lung area without radiographic abnormalities (non-inv-PSB) in 39 MV patients with suspected pneumonia. The comparison of bacterial titers of inv-PSB and non-inv-PSB cultures did not disclose significant differences. Agreement regarding the diagnosis of pneumonia according to recommended diagnostic threshold was observed in 34 of 39 patients (87.1 percent). These results which are in accordance with the pathophysiology of ventilator-associated pneumonia and histologic studies do not support the need to select a certain lung segment for bronchoscopic sampling in most MV patients with suspected pneumonia. PMID- 8417888 TI - Effect of protracted dobutamine infusion on survival of patients in cardiogenic shock treated with intraaortic balloon pumping. AB - The survival of subjects with postmyocardial infarction cardiogenic shock treated with intra-aortic balloon pumping (IABP) differs significantly among various reports. Differences in the criteria for IABP application and in the timing of its initiation have been considered as the main reasons for variations in survival. This study examines whether the way patients in cardiogenic shock are treated prior to IABP may affect their survival. Fifty-five patients in severe postmyocardial infarction cardiogenic shock were classified into three groups according to the rate of dobutamine infusion prior to IABP: the "nondobutamine" (group A, n = 31), the "high-dose dobutamine" (8 to 20 micrograms.kg-1.min-1, group B, n = 17), and the "low-dose dobutamine" (up to 7 micrograms.kg-1.min-1, group C, n = 7). All subjects seen from 1978 to 1983 were recruited for group A, from 1986 to 1990 for group B, and in years 1984, 1985, and 1991 for group C, without using any other classification criteria. It was shown a posteriori that the three groups did not differ in the features of the subjects, in the severity of shock, and in the time length between onset of shock and pumping initiation. None of the 17 subjects of group B could survive under pumping, while 10 of the 31 subjects in group A and 4 of the 7 subjects in group C were weaned off pumping. CONCLUSIONS: A protracted, high-dose pre-IABP administration of dobutamine may adversely affect the survival of patients with postmyocardial infarction cardiogenic shock. PMID- 8417889 TI - Myocardial sarcoidosis. PMID- 8417890 TI - The clinician's perspective on parapneumonic effusions and empyema. AB - Respondents at an interactive symposium on pleural space infections (n = 339) at the 1991 American College of Chest Physicians Annual Scientific Assembly recorded their personal management preferences for hypothetical patients with empyema. The group's preference was to treat pleural sepsis from an anaerobic multiloculated empyema by pleural decortication (49 percent); however, open thoracotomy with directed chest tube placement (22 percent), chest tube placement with intrapleural streptokinase (14 percent), placement of a single chest tube into the largest pleural loculus (8 percent), and placement of multiple small-bore catheters with computed tomographic guidance (7 percent) all had proponents. In the case of a multiloculated empyema not completely drained by a first chest tube in a nontoxic patient, the preference was drainage by a second chest tube, either a small-bore (42 percent) or a large-bore (36 percent) tube. The heterogeneity of responses suggests that prospective trials comparing treatment modalities are needed. PMID- 8417891 TI - Chest radiograph--a poor method for determining the size of a pneumothorax. AB - The ability of chest radiographs to determine the size of a pneumothorax was tested in 16 patients using computed tomographic (CT) scan as a reference method. To determine if CT with a slice thickness of 12 mm could be used, its accuracy was assessed in a lung model experiment. The lung model consisted of a water filled plastic bag (lung) fitted into a plastic chamber (hemithorax), both of approximately the same size and shape as in man. Water was drawn off in incremental steps and a CT was done after each step. The area of the pneumothorax was calculated by computer and when multiplied by slice thickness and number of slices, the total volume of the pneumothorax could be compared with the extracted amount of water. A good correlation (r = 0.99), with the line of regression close to the line of identity, was found between the CT investigation and the artificial pneumothorax. In the patients, the size of the pneumothorax, judged by radiograph using two different methods of calculation, was correlated to the size obtained by CT. The correlation was poor (r = 0.71) irrespective of method of calculation. The size of the pneumothorax estimated by CT showed a good correlation (r = 0.99) to the initial aspirated air volumes in 12 of the 16 patients treated with drainage. A cautious attitude toward the use of chest radiographs for calculations of the degree of lung collapse in patients with pneumothorax is recommended. PMID- 8417892 TI - Vomiting, abdominal pain, and visual disturbances in a 31-year-old man. PMID- 8417893 TI - Human syngamosis. Two cases of chronic cough caused by Mammomonogamus laryngeus. AB - Parasites of the genus Mammomonogamus may occasionally affect the human respiratory tract, causing human syngamosis. We describe two cases of chronic unproductive cough caused by Mammomonogamus laryngeus that occurred in the state of Sao Paulo, Brazil. Some aspects related to human parasitism, diagnostic approach, and treatment are discussed. PMID- 8417894 TI - Continuous positive airway pressure in COPD patients in acute hypercapnic respiratory failure. AB - We used mask continuous positive airway pressure (CPAP) in seven patients with acute hypercapnic respiratory failure in an attempt to avoid endotracheal intubation and mechanical ventilation. Mask CPAP was started at 5 cm H2O and then increased to a maximum of 10 cm H2O depending on the clinical response. In five patients, CPAP significantly improved gas exchange; four of these patients were weaned to oxygen by face mask without needing intubation. In two patients, gas exchange deteriorated even with CPAP of 10 cm H2O. No barotrauma or adverse hemodynamic effects were associated with CPAP. We conclude that a trial of mask CPAP may be warranted before intubation of an alert, acutely hypercapnic patient with COPD. PMID- 8417895 TI - A persistent pulmonary lesion following chemotherapy for metastatic choriocarcinoma. PMID- 8417896 TI - Thoracoscopic implantation of the implantable cardioverter defibrillator. PMID- 8417897 TI - The role of coronary arteriography in demonstration of mural thrombosis after angioplasty. Insights from an experimental model. AB - Although intracoronary thrombosis often occurs after angioplasty and may affect its outcome, the accuracy of arteriography for identification of mural thrombi is unclear. This study analyzed the relationship between arteriographic abnormalities immediately before death and the histologic extent of thrombosis in 77 dogs submitted to balloon injury of intact left anterior descending coronary arteries. Survival time after angioplasty was 120 min. The incidence of mural thrombosis, defined on serial histologic sections, was 65.0 percent. A positive diagnosis of intracoronary thrombus at arteriography (AT+) was based on the presence of any of the following signs: filling defects, retention of contrast material, and slowed or interrupted flow. Seventeen dogs were AT+, and 60 were AT . The overall sensitivity of arteriography was 34 percent, and the specificity was 100 percent. Even considering as significant only thrombi greater than 25.0 percent of the arterial lumen area, 11 of 27 dogs were AT- despite thrombus sizes between 27 percent and 75 percent of lumen area (sensitivity, 59 percent); arteriography consistently missed smaller thrombi (22 of 23 dogs were AT-). Arterial diameters and balloon-induced injury were similar between AT- and AT+ dogs. Scanning electron microscopy depicted a fibrin-poor thrombus in 14 of 19 AT+ dogs and a fibrin-rich thrombus in five, whereas all seven AT+ dogs had fibrin-rich thrombi. Logistic regression analysis showed a correlation between thrombus size and arteriographic positivity, whereas the presence of fibrin and slowed flow of contrast material did not independently predict positive arteriographic results. Thus, arteriography is inaccurate for identification of mural thrombosis after angioplasty, mostly because of its poor sensitivity. PMID- 8417898 TI - Interruption of the aortic arch with associated cardiac anomalies. Survival to adulthood. AB - Interruption of the aortic arch is a rare and usually lethal cardiac malformation. We report a rare case of a patient with IAA distal to the left subclavian artery associated with double outlet right ventricle, subaortic VSD and patent DA who survived to adulthood. In this patient, the complete diagnosis was made by cardiac catheterization and angiocardiography. We discuss the probable mechanisms, diagnostic problems and therapeutic implications of such long survival. PMID- 8417900 TI - Candida endocarditis. A treatable form of pacemaker infection. AB - Fungal endocarditis is a rare complication of permanent pacemaker implantation. In all reports we have identified, this infection has been fatal, diagnosed postmortem. We present a patient in whom early echocardiographic diagnosis resulted in curative surgical and antimicrobial therapy. Fungal endocarditis is an unusual, but treatable complication of permanent pacemakers. PMID- 8417899 TI - From swirling to a mobile, pedunculated mass--the evolution of left ventricular thrombus despite full anticoagulation. Echocardiographic demonstration. AB - We describe the unusual evolution of a left ventricular thrombus following acute anterior myocardial infarction despite adequate anticoagulation. Serial echocardiographic examinations demonstrated the evolution from swirling in the left ventricle through a solid apical mass gradually dislodging into a mobile, pedunculated mass that was removed surgically to prevent embolization. This report emphasizes the need to follow echocardiographically left ventricular thrombi during treatment with anticoagulants, and to identify morphologic changes that may predict embolization. This case suggests that left ventricular thrombectomy should be considered in selected patients in whom a very high-risk thrombus morphology is detected. PMID- 8417901 TI - Diagnosis of paraesophageal omental hiatal hernia by magnetic resonance imaging. AB - We present a case of an enlarging retrocardiac mass lesion in which we observed the magnetic resonance imaging (MRI) finding of a fatty tumor with contiguous blood vessels extending from the abdominal portion into the thoracic portion of the tumor. These surgically verified findings provide an MRI indication of the presence of a paraesophageal omental hiatal hernia. PMID- 8417902 TI - Left ventricular aneurysm, normal coronary arteries and embolization in a patient with systemic lupus erythematosus. AB - We describe a 30-year-old woman with a normal appearing coronary artery associated with SLE who suffered myocardial infarction with left ventricular aneurysm and systemic embolization including cerebral infarctions and saddle embolism. The patient was surgically treated with good results. To our knowledge this is the first reported case of systemic embolism due to myocardial infarction occurring in SLE with a normal coronary artery. PMID- 8417904 TI - Diagnosis of circumferential dissection of the ascending aorta by transesophageal echocardiography. AB - A 28-year-old woman with Marfan's syndrome presented with chest pain; transesophageal echocardiography showed circumferential dissection of the ascending aorta. Both aortic angiography with digital subtraction and computed tomography scanning with contrast were negative for dissection. Circumferential dissection of the ascending aorta was confirmed by surgery at which time replacement of the aorta and aortic valve were performed. Transesophageal echocardiography may become the most practical and reliable procedure for the diagnosis of aortic dissection. PMID- 8417903 TI - Postoperative improvement in blood lactate threshold during exercise in a patient with pulmonary arteriovenous fistula. AB - A 13-year-old female patient with a pulmonary arteriovenous fistula (PAVF) of 24 percent shunt fraction (SF) underwent resection of segments 8 and 9 of the left lower lobe. Three months after the operation, the shunt became insignificant and PaO2 was normalized. In terms of the maximum work rate, maximum VO2 and the VO2 blood lactate relationship, this patient showed a remarkable postoperative improvement in the exercise capacity. PMID- 8417905 TI - Unilateral absence of a pulmonary artery. Data from cardiopulmonary exercise testing. AB - A case of congenital, unilateral absence of a left pulmonary artery is described in a patient presenting with exertional dyspnea. Data from cardiopulmonary exercise testing suggest that the mechanism of dyspnea is secondary to a paradoxic elevation of the physiologic dead space to tidal volume ratio (VD/VT) during exercise. PMID- 8417906 TI - Occupational asthma in a pesticides manufacturing worker. AB - A 34-year-old chemical manufacturing worker had new onset of work-related asthma after several years of exposure to the fungicide, captafol. On specific bronchial challenge testing, he demonstrated a marked and persistent fall in FEV1. Cessation of exposure resulted in improved symptoms and pulmonary function. The delay in symptoms after several years of workplace exposure and the dual reaction demonstrated on specific bronchial challenge testing suggest sensitization to some component of technical-grade captafol, but an IgE response was not detected. PMID- 8417907 TI - Unilateral auto-PEEP in the recipient of a single lung transplant. AB - We report a patient who received a right single lung transplant (SLT) for progressive lymphangioleiomyomatosis and required reintubation for postoperative respiratory distress. She developed hemodynamic instability due to mediastinal shift from unilateral auto-PEEP with hyperinflation of the native lung. Placement of a double lumen endotracheal tube (DLET) and institution of differential lung ventilation restored equal lung inflation and hemodynamic stability. PMID- 8417908 TI - Central nervous system aspergillosis following steroidal therapy for allergic bronchopulmonary aspergillosis. AB - A 52-year-old man with probable allergic bronchopulmonary aspergillosis (ABPA) was treated with adrenal corticosteroids. He subsequently developed central nervous system aspergillosis that was treated successfully with surgery and amphotericin B. Invasive aspergillosis is a rare complication following steroid therapy for ABPA. PMID- 8417910 TI - Rheumatoid nodules in the trachea. AB - We describe the first case of endotracheal rheumatoid nodules in a 45-year-old Chinese man with rheumatoid arthritis. He developed exacerbation of polyarthritis, new subcutaneous nodules and fibrosing alveolitis. At bronchoscopy, four whitish nodules were detected at the tracheal wall, and biopsy showed fibrinoid necrosis, palisaded inflammatory cell infiltrate and vasculitis, consistent with rheumatoid nodules. Both tracheal and subcutaneous nodules regressed with orally administered prednisone. PMID- 8417909 TI - Daytime sleepiness, snoring, and obstructive sleep apnea. The Epworth Sleepiness Scale. AB - The Epworth Sleepiness Scale (ESS) is a simple questionnaire measuring the general level of daytime sleepiness, called here the average sleep propensity. This is a measure of the probability of falling asleep in a variety of situations. The conceptual basis of the ESS involves a four-process model of sleep and wakefulness. The sleep propensity at any particular time is a function of the ratio of the total sleep drive to the total wake drive with which it competes. ESS scores significantly distinguished patients with primary snoring from those with obstructive sleep apnea syndrome (OSAS), and ESS scores increased with the severity of OSAS. Multiple regression analysis showed that ESS scores were more closely related to the frequency of apneas than to the degree of hypoxemia in OSAS. ESS scores give a useful measure of average sleep propensity, comparable to the results of all-day tests such as the multiple sleep latency test. PMID- 8417912 TI - Maloprim-induced pulmonary eosinophilia. AB - A 47-year-old woman developed pulmonary eosinophilia from the use of maloprim as malaria prophylaxis. The diagnosis was confirmed by bronchoalveolar lavage (BAL) and transbronchial lung biopsy. Her condition improved with drug withdrawal and steroid therapy. With the increased use of pyrimethamine and dapsone in the treatment of human immunodeficiency syndrome (HIV) infection, this form of drug allergy may become more common. PMID- 8417911 TI - Hypersensitivity pneumonitis and airways hyperreactivity induced by occupational exposure to penicillin. AB - A patient with penicillin-induced hyperreactive airways in association with hypersensitivity pneumonitis is described. Patch tests and intradermal tests to penicillin were positive. Bronchoalveolar lavage demonstrated a relative lymphocytosis and mild neutrophilia. Symptoms and physiologic abnormalities of pulmonary function and gas exchange resolved on cessation of exposure to penicillin. PMID- 8417913 TI - Disseminated pneumocystosis presenting as a pleural effusion. AB - Extrapulmonary pneumocystosis recently has been reported in a number of tissues. Most cases occurred in patients receiving aerosolized pentamidine prophylaxis. We report a case of disseminated pneumocystosis presenting as a large pleural effusion without apparent lung involvement where Pneumocystis carinii was the only pathogen identified. The absence of parenchymal lesions on chest x-ray film, the lack of hypoxemia and the minimal uptake of gallium all argue against significant lung involvement. The patient was successfully treated with chest tube drainage, intravenous and inhaled pentamidine and orally administered dapsone and trimethoprim. The addition of inhaled pentamidine to intravenously administered pentamidine may have increased pleural fluid levels substantially and its use coincided with the patient's improvement. PMID- 8417914 TI - H-type tracheoesophageal fistula and congenital esophageal stenosis. AB - An 18-month-old boy was seen in the pediatric pulmonary clinic with a history of wheezing, stridor and intolerance to solid foods. Barium esophagram revealed distal esophageal stenosis and subsequently an H-type TEF at surgery. Following the surgery to repair both lesions the child continues to do well. PMID- 8417915 TI - Occupational asthma caused by pectin inhalation during the manufacture of jam. AB - We report a case of pectin-induced occupational asthma in a 35-year-old man. His job involved mixing powdered pectin into a fruit puree during the manufacture of jam. Within minutes of adding pectin, he developed coryza, rhinorrhea, coughing, and wheezing. His symptoms cleared during weekends while away from work and improved with the use of a protective face mask at work. Peak flow rates were significantly lower while at work compared with those at home, and a prick skin test with the pectin powder was positive. We conclude that pectin should be added to the list of the substances known to induce occupational asthma. PMID- 8417916 TI - Delayed pulmonary perforation. A rare complication of tube thoracostomy. AB - Tube thoracostomy is a standard therapy for a number of pulmonary disorders. The procedure is associated with a certain incidence of morbidity related to the technique of insertion, the patient population selected, and the length of time the tube remains in place. Complications of tube placement previously described include empyema, residual pneumothorax, lung perforation, placement of the tube in the chest wall, diaphragmatic perforation, perforation of intraabdominal organs (such as spleen, liver and stomach), unilateral pulmonary edema, bronchopleural fistula, hemothorax, cardiogenic shock and Horner syndrome. A case of a delayed pulmonary perforation developing several days after placement of a chest tube is described with a discussion of the clinical and radiographic findings associated with this complication. A possible pathophysiologic mechanism by which this complication may have occurred is proposed. PMID- 8417917 TI - Respiratory dyskinesia presenting as acute respiratory distress. AB - Incapacitating respiratory distress was the presenting manifestation of a choreiform movement disorder. Because the patient also had asthma, respiratory distress was at first mistakenly attributed to this condition. Despite vigorous asthma management, there was no improvement. However, once the neurologic condition was recognized, use of specific therapy (haloperidol and reserpine) resulted in rapid and sustained remission of respiratory symptoms. PMID- 8417918 TI - Wegener's granulomatosis presenting as a primary seizure disorder with brain lesions demonstrated by magnetic resonance imaging. AB - Since the earliest attempt by Klinger in 1931 to describe the systemic vasculitis subsequently characterized in 1936 by Wegener as Wegener's granulomatosis, this disorder has been reported to present in a number of differing fashions. No previous description relates Wegener's presenting as a seizure disorder, and no reports of MRI of the brain in such patients exists. We relate such a case, along with MRI findings, prior to and after treatment, with a review of neurologic manifestations of the disorder. PMID- 8417919 TI - Gallbladder wall thickening. A new diagnostic sign of acute pulmonary embolism? PMID- 8417920 TI - Absence of IgM and IgG antibody response to the three components of the Mycobacterium bovis BCG 85 complex antigen after BCG vaccination. PMID- 8417921 TI - Salicylate-induced pseudosepsis syndrome. PMID- 8417922 TI - Dysphagia as a complication of oral anticoagulation therapy. PMID- 8417923 TI - Asthma and work. PMID- 8417924 TI - Etiology and diagnosis of pneumonia requiring ICU admission. PMID- 8417925 TI - Incorrect use of metered dose inhalers by medical personnel. PMID- 8417926 TI - Pulmonary parenchymal perforation as a complication of placement of a nasoenteric tube. PMID- 8417927 TI - Autologous "blood patch" pleurodesis for persistent pulmonary air leak. PMID- 8417928 TI - Substitution of metered-dose inhalers for hand-held nebulizers. PMID- 8417929 TI - The metered-dose inhaler supersedes the jet nebulizer. PMID- 8417930 TI - Cough syncope induced by enalapril. PMID- 8417931 TI - Primary pericardial liposarcoma presenting with cardiac tamponade and multiple organ metastases. PMID- 8417932 TI - Pulmonary rehabilitation that includes arm exercise reduces metabolic and ventilatory requirements for simple arm elevation. AB - Simple arm elevation results in increased metabolic and ventilatory requirements in patients with chronic airflow obstruction (CAO). These demands contribute to the dyspnea that is frequently reported when these patients perform activities of daily living involving the arms. We hypothesized that a comprehensive pulmonary rehabilitation (PR) program that includes upper extremity training would lower the ventilatory requirement for arm elevation. Metabolic and ventilatory responses to 2 min of simple arm elevation were studied in 14 patients with CAO before and after PR. Respiratory muscle strength was determined in 11 patients by measurement of maximal transdiaphragmatic pressure (Pdimax). Oxygen uptake (VO2), carbon dioxide production (VCO2), heart rate (HR), minute ventilation (VE), tidal volume (VT), and respiratory rate were measured at rest with the arms down and during 2 min of arm elevation. Before PR, arm elevation led to significant increases in VO2, VCO2, HR, and VE. After PR, pulmonary function, Pdimax, and resting metabolic and ventilatory parameters with the arms down were unchanged; however, during arm elevation, VO2, VCO2, and VE were significantly less than they were before PR. We conclude that a comprehensive PR program that includes upper extremity exercises leads to a reduction in the ventilatory requirement for simple arm elevation. This type of program may allow patients with CAO to perform sustained upper extremity activities with less dyspnea. PMID- 8417933 TI - Reliability coefficient. Man versus machines. PMID- 8417934 TI - The long-term benefits of outpatient pulmonary rehabilitation on exercise endurance and quality of life. AB - Although it is generally accepted that outpatient pulmonary rehabilitation (OPR) improves exercise performance and quality of life (QOL), it is not well established whether these gains are sustained over time. To evaluate this, we attempted to contact the 71 patients who had completed our 6-week OPR program for follow-up 12-min walking distance (12 MD) and QOL measurements. Fifty-one patients (71.8 percent) returned for follow-up testing 11.0 +/- 6.1 months following OPR. Of these, 19 had participated in a structured post-OPR exercise maintenance (EM) program, while 32 had not (non-EM). The 12 MD increased from 2,300 +/- 611 ft at baseline to 2,789 +/- 622 ft post-OPR, while the QOL (higher is better) increased from 81.2 +/- 21.4 to 104.7 +/- 22.2 over this period (both, p < 0.0001). Although the 12 MD decreased by 10.6 +/- 15.8 percent to 2,539 +/- 803 ft at follow-up (p < 0.001), it remained 10.3 +/- 23.4 percent greater than baseline (p < 0.001). Similarly, despite decreasing by 7.6 +/- 13.6 percent to 96.4 +/- 23.3 (p < 0.005), the follow-up QOL remained 22.8 +/- 35.0 percent greater than baseline (p < 0.005). The post-OPR to follow-up declines in 12 MD and QOL were not significantly different between EM and non-EM patients. Thus, only a portion of the initial improvement in exercise endurance and QOL is lost at follow-up months later. Post-OPR EM did not appear to provide measurable long term advantages. PMID- 8417935 TI - Hemodynamic responses to exercise after lung transplantation. AB - A reduced exercise tolerance, maximum oxygen uptake (VO2max), and anaerobic threshold have been reported after lung transplantation (LT). We prospectively assessed the hemodynamic responses to incremental cycle ergometry before and after LT in eight recipients. All recipients underwent a 6-week formal exercise training program. The VO2max increased after versus before LT (13.4 +/- 0.8 vs 9.2 +/- 0.8 ml/min/kg) (p < 0.01). No transition thresholds by analysis of arterial standard bicarbonate were discerned before LT, while the thresholds after LT were abnormally low (VO2 = 9.4 +/- 0.6 ml/min/kg or 35 +/- 3 percent of predicted maximum VO2). An early rise in arterial lactate was similarly observed after LT. Maximum stroke volume index increased in six of seven patients after versus before LT (51 +/- 4 vs 37 +/- 2 ml/beat/m2) (p < 0.05). Three patients demonstrated an increased mean pulmonary arterial pressure at rest, while pressures during exercise were elevated in six. Pulmonary vascular resistance was mildly elevated after LT but decreased appropriately during incremental exercise and was associated with normal cardiac output responses. We conclude that pulmonary vascular abnormalities occurred during hemodynamic exercise testing in the majority of LT recipients; however, exercise limitation was primarily attributed to cardiovascular limitation or to deconditioning in five of the recipients. In the remaining three, the exercise study was considered to be submaximal by virtue of low peak heart rates. A persistent state of deconditioning may have important implications with respect to exercise training regimens after LT. PMID- 8417936 TI - The effect of theophylline on sleep in normal subjects. PMID- 8417937 TI - Pulmonary function after successful heart transplantation. One year follow-up. AB - Congestive heart failure (CHF) has been associated with the development of restrictive ventilatory abnormalities and decreased pulmonary diffusing capacity. Whether these physiologic changes reflect permanent alterations of lung anatomy or result solely from potentially reversible alterations of lung water is not known. To examine this issue, we reviewed the pulmonary function tests (PFTs) and cardiac catheterization data from recipients of successful heart transplants prior to and 1 year after transplantation. Thirty-eight patients met the inclusion criteria (median age, 52 years). The median duration of symptomatic CHF prior to transplantation was 22 months (range, 3 to 72 months). After transplantation, spirometry revealed an improvement in FEV1 from 75.8 +/- 3.5 to 99.1 +/- 2.8 percent of predicted and FVC from 81.3 +/- 3.7 to 101.6 +/- 3.0 percent of predicted (p < 0.001). The FEV1/FVC ratio remained unchanged at 80 percent. Nonsmokers and former smokers had similar improvements in spirometry after transplantation. The TLC improved from 91.1 +/- 3.3 to 105.5 +/- 2.9 percent of predicted (p < 0.001); this improvement was due to an increase in inspiratory capacity. Diffusing capacity for carbon monoxide was decreased before transplantation and showed a small decline after transplantation from 82.3 +/- 3.2 to 76.8 +/- 2.6 percent of predicted (p < 0.05). After correction of severe CHF by cardiac transplantation, normalization of FEV1, FVC, and TLC can be anticipated. Diffusing capacity, however, may actually decline after transplantation. PMID- 8417938 TI - Use of maximum expiratory flow-volume curve parameters in the assessment of exercise-induced bronchospasm. AB - Exercise-induced bronchospasm (EIB) is often inferred from the reduction after exercise in one arbitrarily selected value derived from the maximum expiratory flow-volume (MEFV) curve (eg, FEV1) on a single test; however, patients with symptoms of EIB not meeting these criteria may risk being undiagnosed. To assess the ability of repeated tests using additional MEFV parameters in identifying EIB prone patients, we investigated the effects of exercise provocation on the MEFV curve on two separate occasions. Of 95 patients with symptoms of EIB, 61 had reproducible exercise-induced changes (< 10 percent intraresponse variation), falling into four patterns: 27 (44 percent) had significantly reduced VC and airflow throughout the MEFV curve; 18 (30 percent) had unchanged VC but decreased airflow throughout the curve; 11 (18 percent) had reduced airflow above 50 percent VC but not below 50 percent VC; and 5 (8 percent) had significant reductions in airflow only at 50 percent VC or below. Of the other 34 subjects, 18 had no apparent response, and 16 responded on only one occasion, making objective assessment of these patients' EIB equivocal. We conclude that for a given individual, failure to meet arbitrary criteria does not rule out EIB. Additionally, a more subjective approach that integrates, among other factors, all routine MEFV curve parameters taken from multiple tests with clinical symptoms and history provide a more accurate assessment of EIB. PMID- 8417939 TI - Outcome of subjects with idiopathic pulmonary fibrosis who fail corticosteroid therapy. Implications for further studies. AB - To evaluate the outcome of subjects with idiopathic pulmonary fibrosis (IPF) whose conditions clinically deteriorate while receiving corticosteroid therapy, we studied 12 of these subjects (7 male, 5 female) who received subsequent therapy with intravenous (IV) pulse cyclophosphamide (CPX). Seven of the 12 study subjects died during the course of therapy. Six of these subjects died of respiratory failure, and one died of cholecystitis. Among those who died, the mean age at diagnosis was 63 years compared with 57 years in those who have continued to survive (p = 0.29). Smoking status and pack-years of cigarette smoking were similar between those subjects who died and those who continue to survive. However, subjects who died received CPX for a mean of 6 months, while subjects still living have received CPX for a mean of 16 months (p = 0.01). Subjects who died were given a CPX a mean of 64 months after the onset of symptoms, compared with a mean of 50 months for subjects who are still alive (p = 0.57). Interestingly, there were no significant differences in measures of pulmonary function between living and dead subjects. In fact, measures of lung function and gas exchange remained stable in both groups throughout the period of observation. These data suggest that (1) measures of lung function may not be a reliable indicator of patient mortality in end-stage IPF, and (2) while not statistically significant, these data raise the possibility that duration of symptomatic disease may play a role in the outcome of IPF patients receiving alternative therapeutic agents after failure of corticosteroid therapy. In future intervention trails, controlling entry criteria for duration of disease may prove helpful in determining the effects of these agents on the disease process. These data do not permit a determination of the effect of CPX in patients with IPF. PMID- 8417940 TI - Pasteurella multocida pneumonia in a man with AIDS and nontraumatic feline exposure. AB - A case of acute pneumonia due to Pasteurella multocida ssp multocida occurred in a young man with AIDS and chronic sinusitis. The pneumonia was diagnosed by bronchoscopy and responded to treatment with aztreonam. Epidemiologic investigation revealed the case was temporally related to nontraumatic exposure to cat secretions that the patient presumably had acquired via an aerosol. The cat's oral cavity was cultured and an isolate of P multocida ssp multocida with identical biochemical reactions, DNA restriction patterns, and nearly identical fatty acid profile to that of the patient's isolate was obtained suggesting they were identical strains and therefore epidemiologically linked. A control strain with identical biochemical reactions and antibiotic sensitivities exhibited different patterns. To our knowledge, this is the first such reported infection in a patient infected with human immunodeficiency virus. PMID- 8417941 TI - Right ventricular function at rest and during exercise in chronic obstructive pulmonary disease. Comparison of two radionuclide techniques. AB - Right ventricular function was assessed in 24 patients with COPD, at rest and during submaximal exercise, using both technetium-99m (99mTc) blood-pool and krypton-81m (81mKr) equilibrium ventriculography. Technetium-99m right ventricular ejection fraction (RVEF) at rest was lower than 81mKr RVEF (0.39 +/- 0.12 and 0.54 +/- 0.08, respectively; p < 0.001). During submaximal exercise, there was no increase in RVEF using either imaging technique. This observation contrasted with an increase in RVEF in a group of age-comparable normal subjects during modest submaximal exercise. An inability to obtain spatial separation of right heart structures using 99mTc imaging leads to a value for RVEF that is consistently lower than that measured using 81mKr ventriculography. Resting RVEF is well preserved at rest in most patients with COPD. In contrast to normal subjects, many show an inability to augment right ventricular function during exercise that may contribute to the reduced exercise capacity observed in these patients. PMID- 8417942 TI - Cardiac index vs oxygen-derived parameters for rational use of dobutamine in patients with congestive heart failure. AB - In patients with congestive heart failure (CHF), catecholaminergic agents may exert thermogenic effects that limit their beneficial effects in terms of global tissue oxygenation. Oxygen extraction ratio (O2ER) or mixed venous blood saturation (SvO2) might take into account better than cardiac index (CI) the resultant effect of such agents on peripheral oxygenation. We tested this hypothesis in a series of 20 patients with severe CHF and normal blood lactate levels undergoing pulmonary artery catheterization and receiving incremental doses of dobutamine: 0 (Do), 5 (D5), 10 (D10), and 15 micrograms/kg/min (D15). A significant dose-effect relationship (p < 0.01) was found between dose of dobutamine and CI (CI = 0.06 dose + 1.82). By contrast, no dose-effect relationship was observed between dobutamine dose and either O2ER or SvO2. Indeed, a biphasic profile was observed for O2ER and SvO2. From D0 to D10, O2ER decreased (from 45 +/- 6 to 35 +/- 7 percent) and SvO2 increased (from 52 +/- 7 to 62 +/- 7 percent). From D10 to D15, no further change was observed for both parameters. This latter finding was related to a significant increase in VO2 at D15 (p < 0.01). In these normolactatemic patients with unchanged VO2 from D0 to D10 while DO2 linearly increased (from D0 to D15), the increase in VO2 at D15 was probably due to an increase in oxygen demand induced by the drug. Our results suggest that dobutamine at a dose of 15 micrograms/kg/min can induce an increase in O2 demand that might offset the improvement in CI. Thus, in patients with CHF, oxygen-derived parameters such as O2ER and SvO2 should be more appropriate than CI to assess the efficacy of dobutamine. PMID- 8417943 TI - Postcardiopulmonary bypass lung edema. A preventable complication? AB - Potential etiologic factors of postcardiopulmonary bypass (post-CPB) lung edema (LE) were investigated (phase 1 study). Further patient treatment was modified according to the results of phase 1 study and the influence that these changes had on the incidence of the complication was assessed (phase 2 study). In phase 1 study, among 100 patients who underwent coronary bypass graft surgery, prolonged assisted ventilation was required for severe LE in 7 cases, and 6 patients had a moderate form of LE. Patients who suffered from severe LE had left and right ventricular dysfunction and normal pulmonary vascular resistance. Three predictors of LE were evidenced by logistic regression analysis: number of bypass grafts completed with internal mammary arteries (p = 0.013), transfusions of blood collected in mobile units (p = 0.014), and the combination of a significant lesion of the left main stem and the right coronary artery at preoperative angiography (p = 0.040). In phase 2 study, a further group of 100 patients was treated differently by improving myocardial protection during surgery, achieving a higher rectal temperature at the end of CPB, and reducing the amount of transfusions. This resulted in only one case of LE (p < 0.001). In conclusion, the cases of post-CPB LE evidenced in our study were related to postoperative ventricular dysfunction and blood transfusions. Any means of improving postoperative hemodynamic stability and reducing the need for transfusions could thus prevent the latter complication. PMID- 8417944 TI - Prolonged independent lung respiratory treatment after single lung transplantation in pulmonary emphysema. AB - Single lung transplantation (SLT) is now successfully used in patients with severe emphysema. Mechanical imbalance between the native emphysematous and the healthy transplanted lung can be easily managed, unless severe graft failure occurs, leading to acute respiratory failure. Emergency retransplantation has been used in this setting, since the conventional approach to adult respiratory distress syndrome (ARDS) (mechanical ventilation and positive end-expiratory pressure [PEEP]) fails, due to the mechanical discrepancy between the two lungs. We describe two cases of severe graft failure following SLT in emphysema patients that were successfully treated with prolonged independent respiratory treatment. Mechanical ventilation and PEEP were applied to the failing transplanted lung while the native emphysematous lung was maintained on spontaneous breathing to avoid hyperexpansion and barotrauma. The independent lung respiratory treatment lasted 35 and 25 days, respectively: to our knowledge, these are among the longest-lasting independent respiratory treatments reported. The management was simplified by the early use of a double-lumen tracheostomy cannula as an alternative to orotracheal double lumen tube. PMID- 8417946 TI - Sibling play therapy with homeless children: an opportunity in the crisis. AB - A shelter for the homeless in Hawaii uses the residential setting as an opportunity to provide on-site, flexibly scheduled, culturally sensitive play therapy to young children, specifically targeting siblings as a natural support system to counteract the restricted peer interaction and play experience of this population. Case examples illustrate how sibling groups use the playroom to manage family crises as they happen, resolve conflicts, and express difficult feelings, making sense of their unpredictable world and mastering crucial developmental tasks. PMID- 8417945 TI - Children's perceptions of genital examinations during sexual abuse evaluations. AB - Children's reactions to the medical evaluation of sexual abuse and the methods that enhance their coping ability have not been well addressed in the literature. For many children, a genital examination can be highly stressful, and may even trigger memories of the sexual abuse itself. Stress can be reduced by preparing the child for the examination, by giving the child greater control, and by debriefing the child (and parents) afterward. Research is needed to develop the most effective techniques for reducing children's stress during a genital examination. PMID- 8417948 TI - Hyaluronic acid regulates the function and distribution of sulfated glycosaminoglycans in bone marrow stromal cultures. AB - Treatment with steroid is necessary for adult human bone marrow-derived stroma to support hemopoiesis in vitro. We have investigated the effect of the steroid methylprednisolone on the glycosaminoglycans produced by adult bone marrow stroma. Methylprednisolone did not significantly alter the amount of sulfated glycosaminoglycans produced but the amount of the nonsulfated glycosaminoglycan, hyaluronic acid (HA), was dramatically decreased by the steroid. Culturing methylprednisolone-treated stroma with exogenous HA reduced the proportion of sulfated glycosaminoglycans incorporated into the cell layer and increased the proportion secreted into the medium. It also reduced the capacity of the stroma to bind and stimulate blast colony-forming cells. In fetal liver stroma, methylprednisolone did not decrease the amount of HA or alter the type of heparan sulfate produced. Thus, the ability of stroma cells to respond to steroid and the amount of HA in the extracellular matrix may vary in different marrow microenvironments. This may have functional consequences regarding their abilities to support hemopoiesis. PMID- 8417947 TI - Direct alteration of erythrocyte membrane properties by 1-chloro-2,4 dinitrobenzene without oxidant challenge. AB - The electrophilic agent, 1-chloro-2,4-dinitrobenzene (CDNB), has been widely used as an intracellular glutathione-depleting agent. However, its possible effect on the functional integrity of cell membrane has largely been neglected. Incubation of human erythrocytes (RBC) with various concentrations of CDNB (0.5 to 5.0 mM) in potassium-free, phosphate buffered saline containing ouabain resulted in a drastic depletion of cellular glutathione as well as a dose-dependent increase in passive potassium leakage. Further, an osmotic gradient ektacytometry profile indicated that the deformability index (DI) of CDNB-treated RBC was substantially lower than the DI value of the control. Also, CDNB caused a dose-dependent increase in the rate of shear-induced fragmentation of resealed ghost prepared from treated, intact erythrocytes. These CDNB-induced changes were accompanied by stomatocytic transformations as evidenced by scanning electron micrographs. Additional study indicated that CDNB caused a dose-dependent decrease in thiol concentrations of RBC membrane. SDS-PAGE analysis of membrane proteins revealed new Coomassie blue stainable bands, most noticeable below band-7 (M.W. 20,000). The effects of CDNB on RBC deformability and membrane proteins were also investigated under an atmosphere without oxygen (under nitrogen) and similar effects were observed between that under room air and that under nitrogen. Taken together, these data strongly indicate that CDNB has an adverse effect on the RBC membrane integrity in addition to its ability to deplete intracellular glutathione, possibly through its interaction with membrane sulfhydryl groups. PMID- 8417949 TI - Continuous intravenous administration of rmGM-CSF enhances immune as well as hematopoietic reconstitution following syngeneic bone marrow transplantation in mice. AB - Lethally irradiated Balb/c mice injected with syngeneic bone marrow cells received recombinant murine granulocyte/macrophage colony-stimulating factor (rmGM-CSF) by continuous intravenous infusion for 4 days. When transplanted with 10(5) marrow cells, treated mice showed higher survival (62% compared with 30% in the control group, p < 0.001) and significantly enhanced hematopoietic recovery manifested by 11-fold increase in the peripheral white blood cell (WBC) count. Day 7 marrow from rmGM-CSF-treated mice resulted in 70% survival in lethally irradiated secondary recipients, while marrow harvested under identical experimental conditions from saline-treated mice had no reconstituting capacity at all. When mice were injected with 10(4) marrow cells, 20% of rmGM-CSF treated mice survived as compared with none in the controls. In vitro preincubation of 10(5) and 5 x 10(5) fresh bone marrow cells with rmGM-CSF prior to transplant significantly improved survival of lethally irradiated mice in comparison with control (12% and 37.5% respectively, p < 0.001). Proliferative responses of lymphocytes obtained from rmGM-CSF-treated mice to mitogens and allogeneic C57BL6 splenocytes as well as non-MHC restricted cytotoxicity against tumor cells were significantly higher in rmGM-CSF-treated mice as compared with controls (p < 0.01). These data suggest that a short course of continuous intravenous infusion of rmGM-CSF following BMT or in vitro culturing of bone marrow cells with rmGM CSF improves marrow reconstituting capacity. The mechanism may be by enhancing proliferation and function of committed and perhaps even the more primitive progenitor cell. PMID- 8417950 TI - The effect of nordihydroguaiaretic acid, an inhibitor of prostaglandin and leukotriene biosynthesis, on hematopoiesis of gamma-irradiated mice. AB - The effects of the inhibition of the cyclooxygenase and lipoxygenase metabolic pathways of arachidonic acid on the postirradiation recovery of hematopoietic functions in mice were investigated. Nordihydroguaiaretic acid (NDGA), an inhibitor of prostaglandin (PG) and leukotriene (LT) production, was given to animals in single doses (0.015 to 0.75 mg/mouse) 1 hour before 5 Gy of total-body gamma-irradiation. Enhanced hematopoietic recovery in terms of exogenous and endogenous spleen colonies, femoral granulocyte-macrophage colony-forming cells and peripheral blood granulocyte levels was observed at higher doses of NDGA. The treatment used influenced neither lymphocyte nor erythrocyte postirradiational levels or hemoglobin concentration. A comparison of the effects induced by a high dose of NDGA (0.3 mg per mouse) with those observed after an isomolar dose of indomethacin (an inhibitor of PG production) indicated only slight differences between these two drugs. An isomolar dose of esculetin (an inhibitor of LT production) had no effect on the postirradiation behavior of hematopoiesis. The results suggest that the inhibition of PG production plays the main role in the mechanism of NDGA action. Inhibition of LT production seems to be of less importance for hematopoiesis in these in vivo conditions. PMID- 8417951 TI - Mechanism of radioprotection conferred by the immunomodulator AS101. AB - AS101 [ammonium trichloro (dioxyethylene-0-0') tellurate] is a new synthetic compound previously described by us as having immunomodulating properties and minimal toxicity. Clinical trials are currently in progress with AS101 in AIDS and cancer patients. AS101 has recently been found to have radioprotective effects on hemopoiesis in irradiated mice when administered prior to irradiation. Since the early progenitors, spleen colony-forming units (CFU-S), are the critical cells needed for the reconstitution of the hemopoietic system, the mechanisms of action of AS101 were explored in this study by examining the compound's effect on the recovery of CFU-S, its protective effect on endogenous CFU-S and its effect on self-renewal of CFU-S. We also studied the effect of AS101 on the induction of progenitor cells into the radioresistant S-phase of the cell cycle. On days 1 and 5 after irradiation, the number of CFU-S in the bone marrow and spleen of AS101-treated mice was significantly higher than that of PBS injected mice. Nine days after sublethal doses of irradiation, the number of endogenous spleen colonies was highest in mice given AS101 every 24 hours or every other day for 1 week prior to irradiation. AS101 administered immediately after irradiation, however, also resulted in an increase in the endogenous CFU-S. The higher number of CFU-S found in each 9-day endogenous spleen colony suggests increased self-renewal of CFU-S in AS101-treated mice. Moreover, we found that AS101 induced a higher number of progenitor cells in the S-phase of the cell cycle. These findings suggest that the radioprotection conferred by AS101 results from induction of progenitor cells in DNA synthesis (S-phase) and from the enhanced stimulation of CFU-S, not only toward proliferation but also toward CFU S self-renewal. PMID- 8417952 TI - Estrogen blocks the cimetidine-induced suppression of CFU-GM. AB - Cimetidine, an H2-histamine antagonist used for the treatment of duodenal ulcers, has been shown to suppress granulocyte/macrophage colony forming cells (CFU-GM) in males. This study was initiated to examine the role of sex hormones on this cimetidine-induced suppression of CFU-GM. Preincubation of light-density, nonadherent bone marrow cells in male patients with 10(-6) M testosterone resulted in a modest decrease in the suppressive effect of cimetidine, whereas preincubation with 10(-6) M 17-beta-estradiol, for as little as 10 minutes, completely abolished the 50% reduction in colony numbers induced by cimetidine. Using supra-pharmacologic doses of cimetidine in order to detect CFU-GM suppression in female patients, identical results were obtained. Tamoxifen completely reversed this protective effect of estrogen and preincubation with hydroxyurea and the elimination of T cells from the system failed to alter any of these results, lending support to the likelihood that both cimetidine and estrogen directly affect marrow myeloid progenitor cells. PMID- 8417953 TI - Erythroid potentiating activity of tissue inhibitor of metalloproteinases on the differentiation of erythropoietin-responsive mouse erythroleukemia cell line, ELM I-1-3, is closely related to its cell growth potentiating activity. AB - The erythroid-potentiating activity (EPA) of the tissue inhibitor of metalloproteinase-1 (TIMP-1) was re-examined using ELM-I-1-3, a mouse erythroleukemia cell line, which responded well to erythropoietin. Depletion of pre-existing TIMP-1 from fetal calf serum in culture medium using monoclonal antibody suppressed erythropoietin-induced differentiation as measured by the induction of hemoglobin, commitment assay and globin mRNAs. The removal of TIMP-1 also suppressed the proliferation of ELM-I-1-3 as measured by cell number and de novo DNA synthesis. These changes were reversed by the addition of purified TIMP 1 to the culture medium. Anti-TIMP-1 antibody also blocked both hexamethylene bisacetamide (HMBA)-induced erythroid differentiation and the proliferation of both ELM-I-1-3 and Friend erythroleukemia cells. Considering previous reports analyzing the chemical induction of Friend mouse erythroleukemia cell differentiation, our results suggest that erythropoietin- or HMBA-induced erythroid differentiation might also be coupled with cell proliferation. Our 3H thymidine-uptake experiment shows that TIMP-1 removal was also effective in the inhibition of cell growth of various other cell lines in addition to erythroleukemia cell lines. These results suggest that EPA action of TIMP-1 on erythroid leukemia cell lines is closely related to its activity to promote the cell growth of various cell lines and cells including erythroleukemia cell lines. PMID- 8417954 TI - Modulation of macrophage Fc gamma receptors by rGM-CSF. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a hematopoietic growth factor being used increasingly to support white blood cell counts in hematologic disorders. Since the survival of IgG-sensitized cells following blood transfusions and the clearance of immune complexes are important in these disorders, we investigated the effect of GM-CSF on the Fc gamma receptors largely responsible for this immune clearance. Human monocytes were cultured in buffer or 100 U/mL of recombinant GM-CSF (rGM-CSF) for 48 hours. Flow cytometry was used to evaluate changes in the expression of the three Fc gamma receptors. Fc gamma RII was the only Fc gamma receptor significantly increased by rGM-CSF. This increase in Fc gamma RII surface protein was correlated with an increase in macrophage binding of erythrocytes sensitized with IgG. In addition, an increase in monocyte binding of IgG-sensitized RBCs was observed in RBCs sensitized with murine IgG2b antibody, which preferentially binds to Fc gamma RII. rGM-CSF also increased the monocyte Fc gamma RII-dependent low-affinity binding site for trimeric IgG. Furthermore, rGM-CSF was observed to increase the expression of monocyte Fc gamma RII mRNA, including that for Fc gamma RIIA. Thus, these studies demonstrate that GM-CSF increases monocyte Fc gamma RII expression and function and suggests that a similar process may be present in vivo. This effect may be either beneficial (increased clearance of immune complexes) and/or detrimental (increased transfusion requirements) in select patients. PMID- 8417955 TI - Stimulation of erythroid progenitors by high concentrations of erythropoietin results in normoblasts arrested in G2 phase of the cell cycle. AB - The cell cycle status of human erythroid precursors generated in a two-step liquid culture was studied by a double-labeling flow cytometric technique. Following a first phase, where peripheral blood mononuclear cells were cultured in the presence of a combination of growth factors, not including erythropoietin (Epo), the cells were washed and recultured in a second phase in the presence of Epo. This procedure resulted in a stimulation of the proliferation and maturation of erythroid precursors. In the presence of optimal concentrations of Epo (2 U/mL), a high percentage (> 40%) of cells were found in the S phase of the cell cycle until day 10. Then, as a result of maturation, the proportion of cells in S gradually decreased, reaching less than 2.0% by day 21. At this time, the culture consisted of > 95% hemoglobin-containing, nonproliferating, orthochromatic normoblasts. Cell cycle analysis of this normoblast population demonstrated a bimodal distribution; while the majority of the cells had a diploid (2C) DNA content, i.e., cells in G1 (or G0) phase, a sizable fraction was tetraploid (4C) corresponding to cells in G2. In contrast, in cultures stimulated with physiological concentrations of Epo (around 50 mU/mL), all the terminally differentiated cells were arrested at the G1 phase. These results suggest that Epo is an essential growth-promoting factor for erythroid precursors, but supraphysiological concentrations, such as present in vivo in severe anemia (e.g., aplastic anemia) or after Epo administration, may be associated with development of normoblasts with abnormal DNA content. PMID- 8417956 TI - The "b" mutated gene in heterozygous Belgrade anemic rat. AB - The Belgrade b/b rat has an autosomal recessive mutation which in homozygous state induces severe anemia. So far, this mutation has been considered a recessive one and the heterozygous animals (+/b) as phenotypically normal. In this study, we showed that at the hematologic level, the heterozygous animals acquire some of the anemic characteristics as well. Namely, the young +/b animal displays reticulocytosis of 3.1 +/- 1.0%, identical to b/b rat, compared with 0.8 +/- 0.4% in young normal animals. This conclusion was further supported by examination of beta-globin expression. The level of beta-globin mRNA in anemic and heterozygous reticulocytes is decreased, as estimated by dot blot hybridization, to 25% and 50% of normal level, respectively. Although inapparent phenotypically, b mutated allele disturbs early erythropoiesis and markedly decreases globin mRNA level in the heterozygous rat. PMID- 8417957 TI - Insulin-like growth factors stimulate erythropoiesis in serum-substituted umbilical cord blood cultures. AB - The stimulatory effects of the insulin-like growth factors (IGFs) on erythroid colony formation by hematopoietic progenitor cells derived from adult bone marrow and peripheral blood have been well demonstrated. To enhance our understanding of the potential role of IGFs in human newborn erythropoiesis, we studied the effects of IGF-I and IGF-II on neonatal erythropoiesis in vitro. Erythroid progenitor cells were recovered from umbilical cord blood collected at scheduled Cesarean sections from women with term, uncomplicated pregnancies. Light-density cord blood mononuclear cells were cultured for 7 days in a serum-substituted culture system supplemented with and without purified human recombinant erythropoietin (0.5 to 2.0 U/mL) and recombinant human IGFs. Addition of IGF-I (1 to 100 ng/mL) resulted in up to a 4-fold increase over baseline erythropoietin dependent colony formation at 7 days with a maximal effect at 10 ng/mL. While subphysiologic doses of IGF-II caused a modest stimulation of erythropoiesis, addition of a physiologic concentration (100 ng/mL) resulted in up to a 4-fold enhancement in erythroid colony formation. This stimulatory effect persisted in cultures of mononuclear cells that were depleted of monocytes, myeloid cells, T cells and B cells prior to culture, suggesting that enhancement is the result of a direct IGF effect on progenitor cells. Addition of IGFs in the absence of erythropoietin resulted in erythroid colony formation in some but not all experiments. Whereas IGF-I was the primary regulator of erythropoietin independent erythroid colony formation by adult erythroid progenitors, IGF-II was the predominant regulator of erythropoietin-independent erythroid colony formation by neonatal progenitor cells. We conclude that IGFs in physiologic doses directly stimulate neonatal cord blood erythroid progenitor cells and may play a role in developmental control of human erythropoiesis. PMID- 8417958 TI - IL-1 beta inhibits adipocyte formation in human long-term bone marrow culture. AB - Addition of recombinant interleukin-1 beta (IL-1 beta) to human long-term bone marrow culture (LTBMC) was found to have a profound effect on adipocyte formation. Control cultures developed 133 +/- 58 adipocytes/flask compared with 3 +/- 2 adipocytes/flask in IL-1 beta-treated flasks (mean +/- SEM, n = 6 for both groups). In terms of cell number, morphology and colony forming cell, IL-1 beta did not have significant effects on LTBMC, although IL-1 beta slightly enhanced colony-stimulating activity of LTBMC-conditioned medium. To further analyze the effect of IL-1 beta on adipocytes, stromal cells were passaged to reduce hemopoietic cell content, and marrow fibroblasts were converted to adipocytes following addition of hydrocortisone. In both types of assay, IL-1 beta again reduced adipocyte formation: control 1095 +/- 76 adipocytes/flask, IL-1 beta treated 7 +/- 2 adipocytes/flask (n = 3 for both groups). When preformed adipocytes were treated with IL-1 beta for 1 week, the number of lipid-laden cells decreased to 41 +/- 13% of its value before treatment compared to 87 +/- 9% for untreated controls. The addition of neutralizing concentrations of anti-TNF did not inhibit these effects of IL-1 beta. These results suggest that 1) adipocyte formation is not essential for healthy LTBMC growth, or at least IL-1 beta is an effective substitute for adipocytes; 2) IL-1 beta by itself does not significantly enhance hemopoiesis in LTBMC, and this has implications for clinical trials with this cytokine; 3) IL-1 beta produces major changes in stromal cell development, and 4) overgrowth of fat in marrow may reflect defective cytokine regulation in vivo. PMID- 8417959 TI - Advantages of gradient vs. 5-fluorouracil enrichment of stem cells for retroviral mediated gene transfer. AB - Retrovirally mediated gene transfer into murine totipotent hematopoietic stem cells (THSC) may be more efficient when the donor stem cells are enriched. We have used a rapid, nontoxic density gradient separation of mouse marrow to enrich stem cells. By characterizing the cell types in various fractions of the gradient, we found the majority of the THSC, spleen colony forming stem cells (CFU-S), erythroid burst forming cells (BFU-E) and dividing cells were in the same fraction. The gradient enrichment technique was then compared with one requiring 5-fluorouracil (5-FU) treatment of donor mice prior to marrow harvest. Cells enriched by both methods were tested for their ability to mediate retroviral gene transfer into normal mice. Gradient enrichment provided only one third as many nucleated cells as 5-FU treatment from the same number of donors. During the subsequent 4-day in vitro exposure to the retrovirus and growth factors, however, the number of gradient enriched cells increased 1.6-fold while the number of 5-FU treated cells decreased 3-fold. In lethally irradiated recipients, there was no difference between gradient and 5-FU enriched donor cells in the proportion of cells that generated CFU-S nor in the percentage of CFU-S that were infected. Secondary hosts did show differences. Gradient-enriched cells maintained more survivors for up to 6 months posttransplantation and more of the survivors were positive for the retrovirus. It is clear that the gradient method provides a rapid means to enrich CFU-S and THSC without exposure to the toxic effects of 5-FU. PMID- 8417960 TI - Restorative effect of IL-3 on adherence of cloned hemopoietic progenitor cell to stromal cell. AB - Mammalian hemopoiesis results from a complex interaction between hemopoietic progenitor cells, stromal cells and extracellular matrix components, orchestrated by specific glycoprotein growth factors. Recently, these growth factors have been shown to possess an important function, apart from stimulation of proliferation, and that is suppression of an active cellular process of programmed cell death, or apoptosis. Highly specific biochemical and morphologic changes have been shown to occur during apoptosis, but their reflections on cellular functions are poorly understood. Interleukin-3 (IL-3)-dependent FDCP-1 (factor-dependent cell lines cloned in Paterson Laboratories) cells were studied for their ability to adhere to hemopoietic stroma in a temporal fashion under conditions of apoptosis and following rescue from apoptosis with growth factor. It was found that cloned FDCP 1 cells always maintained, in the presence of a source of IL-3 (either WEHI conditioned medium or rm-IL-3), bound cloned hemopoietic stromal cell GB1/6 in a constant fashion for 20 hours, while cells starved of IL-3 experienced a 50% time dependent decrement in binding. If IL-3 were added back to FDCP-1 that had been starved of growth factor for 8 hours, but not 12 hours, adherence to stroma was restored to that of control cells always in the presence of IL-3. Granulocyte/macrophage colony-stimulating factor (GM-CSF), Interleukin-6 (IL-6) and transforming growth factor-beta (TGF-beta) did not restore cytoadherence. By transmission electron microscopy, nucleus and cytoplasm of IL-3-replenished cells resembled that of control cells. These data indicate that at least some events related to apoptosis were reversible for a period up to 8 hours, but not 12 hours, in cells that had been rescued by readdition of IL-3. These findings offer important insight into a way in which bone marrow progenitor cells may be maintained in a condition that optimizes their ability to engraft stroma during transplantation. PMID- 8417961 TI - Is hyaluronic acid the "organizer" of the extracellular matrix in marrow stroma? PMID- 8417962 TI - Ubiquitous expression of cytokines in diverse leukemias of lymphoid and myeloid lineage. AB - It has recently been suggested that autocrine production of hematopoietic regulatory molecules can modulate the cardinal features of many leukemic states: excessive proliferation of the neoplastic cells and suppression of the normal elements. We therefore analyzed samples obtained from 57 patients with a variety of hematologic malignancies (21, acute myelogenous leukemia; 14, acute lymphoblastic leukemia; 12, Philadelphia chromosome-positive chronic myelogenous leukemia [blast phase] or acute leukemia; 5, chronic lymphocytic leukemia; and 5, chronic myelomonocytic leukemia) for expression of interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) transcripts on Northern blots. TNF alpha mRNA was discerned in almost half of the samples (47%), and was expressed in some patients with every type of leukemia, except T-cell acute lymphoblastic leukemia (ALL). Expression occurred with great frequency in samples (12 of 15 [80%]) from monocytic (acute or chronic) leukemias, and from advanced chronic lymphocytic leukemia (4 of 5 samples [80%]). IL-1 beta transcripts were detected in 20 of 57 samples (35%). Its presence, like that of TNF-alpha, was ubiquitous, and only chronic lymphocytic leukemia and T-cell acute lymphoblastic leukemia cells consistently failed to produce IL-1 beta message. Therefore it appears that TNF-alpha and/or IL-1 beta mRNA can be found in the leukemic cells from a substantial subset of patients with B cell-derived acute lymphoblastic leukemia as well as with chronic and acute myeloid, monocytic or lymphocytic leukemias. Because these cytokines have potent direct and indirect effects on normal and malignant hematopoiesis, their widespread constitutive expression by neoplastic blood cells may play a fundamental role in driving the leukemic process. PMID- 8417963 TI - In vivo and in vitro effects of 3-hydroxypyridin-4-one chelators on murine hemopoiesis. AB - The effects of 3-hydroxypyridin-4-one (HPO) iron chelators and desferrioxamine (DFO) on murine hemopoiesis in vivo and in vitro have been compared in order to investigate the mechanism by which leucopenia in mice and granulocytopenia in man occurs with 1,2-,dimethyl-HPO (CP20). Administration of 60 doses of 200 mg/kg CP20 to Balb/c mice resulted in significant anemia, lymphopenia and granulocytopenia accompanied by bone marrow hypocellularity. DFO and CP94 (1,2,diethyl-HPO) at the same dose also caused lymphopenia but marrow cellularity was unaffected. When marrow from untreated mice was incubated with HPOs and DFO, erythroid burst-forming cells (BFU-E) and granulocyte/macrophage colony forming units (CFU-G+Mac), colony growth was inhibited in a dose-dependent manner at micromolar concentrations. The addition of iron to saturate the chelators abrogated the effects of DFO, but not those of the HPOs. With the HPO-iron complexes, addition of sufficient iron to saturate the transferrin in the medium reversed the inhibitory effects of the relatively hydrophilic CP20-iron complex but not those of the more lipophilic CP94-iron complex. Addition of further iron saturated transferrin also corrected inhibition by the CP94-iron complex. These results show that HPO-iron complexes potentially have antiproliferative effects unlike DFO-iron complex (FO). The difference in the relative effects of CP20 to CP94 on hemopoiesis in vivo and in vitro suggests that additional factors to those inhibiting hemopoiesis in marrow cultures may operate with the long-term administration of iron chelators in vivo. PMID- 8417964 TI - Mixed colony formation composed of erythroblasts and myeloblasts by leukemic blast progenitors in patients with erythroleukemia. AB - To determine the clonal origin and growth requirement of leukemic blast progenitors in erythroleukemia, acute myelogenous leukemia (AML) M6, T cell depleted mononuclear cells obtained from 5 erythroleukemia patients were cultured in methylcellulose media. Although plating efficiency did not significantly differ from those in the other AML subtypes, the morphology of colonies in erythroleukemia was distinct. Two types of colonies were formed; one was composed of myeloblasts, and the other consisted of erythroblasts and myeloblasts. The "mixed" colony formation was not stimulated by interleukin-3, granulocyte macrophage colony-stimulating factor, granulocyte colony-stimulating factor or erythropoietin. Both types of colonies were formed spontaneously in the absence of supplemented growth-stimulatory factor when cultured at high density. The results suggest that leukemic blast progenitors in erythroleukemia originated at the stage of bipotent hematopoietic precursor capable of differentiating into erythroid and myeloid lineages. The formation of mixed colonies composed of erythroblasts and myeloblasts in the absence of erythropoietin or colony stimulating factor may indicate the deranged hematopoiesis in erythroleukemia. PMID- 8417965 TI - The ovalbumin family of serpin proteins. AB - A protein family, the 'Ov-serpins' has been identified by comparing amino acid sequence, protein characteristics and gene organization. The Ov-serpins would not be recognized as a family based on sequence identity alone. This example suggests that combinations of characteristics may need to be examined to identify family groupings within the serpin superfamily. PMID- 8417966 TI - A metal-linked gapped zipper model is proposed for the hsp90-glucocorticoid receptor interaction. AB - In the presence of certain metals, regions of the hormone binding domain of the glucocorticoid receptor (GR) are capable of binding the 90 kDa heat shock protein (hsp90). Using secondary structure prediction methods in correlation with the experimental data, we propose a model which predicts the presence of two widely spaced leucine zipper-like heptads on either side of a central subdomain. The heptads could interact hydrophobically with similar regions on the hsp90 homodimer, bringing putative metal binding residues on each protein close enough to establish a shared metal bridge. The central subdomain between heptads is suggested to contain regions involved in metal binding, steroid binding, and conformational mobility. The hypothetical model that we are proposing therefore addresses the nature of the structural link between hsp90 binding, hormone binding, and conformational changes in the receptor. PMID- 8417967 TI - Low pH crystal structure of glycosylated lignin peroxidase from Phanerochaete chrysosporium at 2.5 A resolution. AB - The heme-containing glycoprotein lignin peroxidase (pI 4.15) has been crystallized at pH 4.0. The structure of the peroxidase from the orthorhombic crystals has been determined by multiple isomorphous replacement. The model comprises all 343 amino acids, one heme molecule, and three sugar residues. It has been refined to an R-factor of 20.3%. The chain fold of residues 15 to 275 is in general similar to those of cytochrome c peroxidase. Despite binding of the heme to the same region and a similar arrangement of the proximal and distal histidine as in cytochrome c peroxidase a significantly larger distance of the iron ion to the proximal histidine is observed. Distinct electron density extending from Asn-257 and at the distal side of the heme indicates ordered sugar residues in the crystal. PMID- 8417968 TI - An improved retroviral vector for assaying promoter activity. Analysis of promoter interference in pIP211 vector. AB - We recently developed a novel promoter assay system using a retroviral vector (pIP200 series). Transcription from the internal promoter, which had been inserted for the promoter assay, was shown to be interfered with by transcription from the upstream long terminal repeat (LTR). Here we report a new high-titer 'self-inactivating' vector, in which transcription interference was virtually eliminated. This new vector was constructed by introducing only a very minor mutation into the 'TATA box' in the 3'-LTR. This mutation was successfully transferred to the 5'-LTR after reverse transcription, yielding a provirus incapable of transcribing viral RNA. The viral titer was not reduced by the mutation, permitting general application of this virus. PMID- 8417969 TI - Loss of alpha-tocopherol upon exposure to nitric oxide or the sydnonimine SIN-1. AB - SIN-1 which spontaneously decomposes to yield nitric oxide (NO.) and superoxide anion (O2.-) radicals caused a loss of microsomal alpha-tocopherol paralleled by the formation of alpha-tocopheryl quinone. The loss was partially prevented by superoxide dismutase but not by catalase. The SIN-1-induced loss of alpha tocopherol also occurred when tocopherol was dissolved in ethanol/potassium phosphate buffer (20/80, v/v). Likewise, addition of authentic NO. to alpha tocopherol dissolved in ethanol resulted in loss of the vitamin and quinone formation. These results suggest that NO. or its products such as peroxynitrite or nitrogen dioxide react with alpha-tocopherol, the quinone derivative being a major oxidation product. Depletion of vitamin E by NO. may contribute to tissue injury, e.g. in neuronal tissues. PMID- 8417970 TI - Urea-generated free rotating water molecules are active in the protein unfolding process. AB - The critical urea concentration (C3*) which destabilizes the structure of bovine serum albumin and chymotrypsinogen was determined by UV difference spectroscopy. The increase of the relative content of mobile rotating water molecules in aqueous urea was formerly shown by millimeter spectroscopy [1]. The rise of rotator content at a urea concentration C3 > or = C3* when the bulk water is practically exhausted is suggested as a main driving force of protein unfolding. PMID- 8417971 TI - Substrate induced changes of the active site electronic states in reduced cytochrome P450cam and the photolysis product of its CO complex. Low-temperature magnetic circular dichroism data. AB - MCD spectra of camphor-free and camphor-bound reduced cytochrome P450cam have been recorded for the near UV and visible spectral regions at temperatures from 300K down to 2.1K and compared with those of the carbon monoxide photoproducts generated at 4.2K. In the absence of camphor, the reduced P450 is spectroscopically different from the photoproduct. In the presence of camphor, however, the spectra of the reduced P450 and of the photoproduct are almost similar and behave like the photoproduct of the camphor-free enzyme. This behavior indicates that substrate binding induces a higher active site rigidity. From the significant alteration of the temperature dependence of the MCD intensity for the reduced enzyme induced by camphor binding it is concluded that the near degeneracy of the electronic ground state in the substrate-free enzyme is removed by substrate binding. PMID- 8417972 TI - Absence of expression of interleukin-6 (IL-6) mRNA in regenerating rat liver. AB - The serum level of IL-6 and expression of IL-6 mRNA in hepatocytes from regenerating liver were investigated in the rat. The IL-6 level in the serum was not significantly different from that of a control group of rats submitted to an acute experimental inflammation. IL-6 mRNA expression did not occur in the liver of hepatectomized rats as judged from Northern blotting experiments using an IL-6 riboprobe. These results suggest that if IL-6 is implicated in hepatic regeneration, this cytokine is not produced by the regenerating liver and must be delivered exogenously to the liver to modulate hepatic regeneration. PMID- 8417973 TI - Expression of a cDNA encoding the glycolipid-anchored form of rat acetylcholinesterase. AB - We amplified by PCR and characterized a fragment of cDNA from rat spleen, encoding the distinctive C-terminal region of the acetylcholinesterase (AChE) H subunit. A recombinant vector encoding this subunit was constructed and expressed in COS cells: the H subunits produced glycophosphatidylinositol (GPI)-anchored dimers, showing that the spleen cDNA fragment contained a functional GPI cleavage/attachment site. Using PCR, we did not detect mRNAs encoding AChE H in rat muscle or hypothalamus. In the liver of 16-day rat embryos, we found both H and T transcripts, in agreement with the presence of both GPI-anchored dimers and amphiphilic monomers of type II. In addition, we detected 'read-through' (R) transcripts, in which regular introns are spliced, but the intervening sequence between the common exon 4 and the alternative exon 5 (H) is maintained. PMID- 8417974 TI - Effect of metal ions on the activity of casein kinase II from Xenopus laevis. AB - Casein kinase II purified from the nuclei of Xenopus laevis oocytes as well as the recombinant alpha and beta subunits of the X. laevis CKII, produced in E. coli from the cloned cDNA genes, were tested with different divalent metal ions. The enzyme from both sources was active with either Mg2+, Mn2+, or Co2+. Optimal concentrations were 7-10 mM for Mg2+, 0.5-0.7 mM for Mn2+ and 1-2 mM for Co2+. In the presence of Mn2+ or Co2+ the enzyme used GTP more efficiently than ATP as a phosphate donor while the reverse was true in the presence of Mg2+. The apparent Km values for both nucleotide triphosphates were greatly decreased in the presence of Mn2+ as compared with Mg2+. Addition of Zn2+ (above 150 microM) to an assay containing the optimal Mg2+ ion concentration caused strong inhibition of both holoenzyme and alpha subunit. Inhibition of the holoenzyme by 400 microM Ni2+ could be reversed by high concentrations of Mg2+ but no reversal of this inhibition was observed with the alpha subunit. PMID- 8417975 TI - The effect of annexin IV and VI on the fluidity of phosphatidylserine/phosphatidylcholine bilayers studied with the use of 5 deoxylstearate spin label. AB - An effect of annexin IV and VI on the fluidity of phosphatidylserine/phosphatidylcholine (PS/PC) membranes was studied by spin labeling technique with the use of 5-doxylstearic acid. It was found that calcium ions at micromolar concentrations induced a marked decrease in the order parameter of PS/PC membranes. This effect was completely abolished by both annexins. The influence of annexins on the mobility of fatty acid chains in the studied region in PE/PC membranes was insignificant. PMID- 8417976 TI - Electrical stimulation partly reverses the muscle insulin resistance caused by tenotomy. AB - It was shown that 15-min electrical stimulation of the rat sciatic nerve greatly increases the in vitro measured sensitivity of lactate formation, glucose transport, and glycogen synthesis to insulin, impaired by previous tenotomy. The insulin sensitivity of all these processes was, however, still below that found in the stimulated intact soleus muscle. Extending the stimulation up to 30 min did not cause any further changes in insulin sensitivity either in tenotomized or in intact muscles. PMID- 8417977 TI - Defensin-6 mRNA in human Paneth cells: implications for antimicrobial peptides in host defense of the human bowel. AB - The epithelial surface of the human small intestine is a barrier between the host and the microbial environment of the lumen. A human small intestine cDNA clone was found to encode a new member of the defensin family of antimicrobial peptides, named human defensin-6. Tissue expression of this mRNA is specific for the small intestine as determined by Northern blot analysis and polymerase chain reaction analysis. In situ hybridization demonstrated that human defensin-6 mRNA localizes to Paneth cells in the crypts of Lieberkuhn. The finding of an abundant defensin mRNA in human Paneth cells supports the notion that these epithelial cells may play a key role in host defense of the human bowel. The results also strengthen the hypothesis that peptide-based host defenses are prevalent at mucosal surfaces in mammals. PMID- 8417978 TI - Identification and differential expression of yeast SEC23-related gene (Msec23) in mouse tissues. AB - We have isolated a yeast SEC23-related clone (Msec23) from mouse fibroblast cDNA library. It has an open reading frame of 1721 bp (64% homologous to SEC23-2.3 kb) which can potentially encode a 64.7 kDa protein (61% homologous to 85.4 kDa product of SEC23). The deduced Msec23 protein (Msec23p) sequence contains three successive Ig-like domains at the N-terminus followed by amphipathic alpha helical regions, suggesting the potential of Msec23p to interact with other protein components. Further, Msec23 is differentially expressed in mouse tissues with its high level in brain and fibroblasts. PMID- 8417979 TI - Regulation of Cu,Zn- and Mn-superoxide dismutase transcription in Saccharomyces cerevisiae. AB - The regulation of Cu,Zn- and Mn-superoxide dismutases (SOD) was investigated by Northern blotting and gene fusions of SOD1 and SOD2 promoters with the beta galactosidase reporter gene. Cu,ZnSOD expression was increased 3-fold under glucose derepressing conditions, and decreased 4- to 6-fold by oxygen or heme deficiency. MnSOD expression was increased 5-fold by glucose derepression, and decreased 8- to 10-fold by anaerobiosis and 4- to 5-fold by heme deficiency. Induction by paraquat was modest, about 50% for SOD1 and 100% for SOD2; it was apparently independent of the respiratory chain function. PMID- 8417980 TI - Base transitions and base transversions seen in mutations among various types of the hepatitis C viral genome. AB - We counted base transitions (Ts) and transversions (Tv) in mutations among various types of hepatitis C virus (HCV) sequences. The Ts/Tv ratio was useful for distinguishing the (geno)type of the compared HCV sequences, since the ratio was 4.0 or more among sequences of the same type, while it was less than 2.0 among sequences of different types. This observation reflects the evolutionary pathway of HCV in which Tv accumulate among distant genomes by mechanisms including multiple base substitutions. PMID- 8417981 TI - Marcus M. Rhoades (1903-1991). PMID- 8417982 TI - Unequal exchange and meiotic instability of disease-resistance genes in the Rp1 region of maize. AB - The Rp1 region of maize was originally characterized as a complex locus which conditions resistance to the fungus Puccinia sorghi, the causal organism in the common rust disease. Some alleles of Rp1 are meiotically unstable, but the mechanism of instability is not known. We have studied the role of recombination in meiotic instability in maize lines homozygous for either Rp1-J or Rp1-G. Test cross progenies derived from a line that was homozygous for Rp1-J, but heterozygous at flanking markers, were screened for susceptible individuals. Five susceptible individuals were derived from 9772 progeny. All five had nonparental combinations of flanking markers; three had one combination of recombinant flanking markers while the other two had the opposite pair. In an identical study with Rp1-G, 20 susceptible seedlings were detected out of 5874 test cross progeny. Nineteen of these were associated with flanking marker exchange, 11 and 8 of each recombinant marker combination. Our results indicate that unequal exchange is the primary mechanism of meiotic instability of Rp1-J and Rp1-G. PMID- 8417983 TI - Do deleterious mutations act synergistically? Metabolic control theory provides a partial answer. AB - Metabolic control theory is used to derive conditions under which two deleterious mutations affecting the dynamics of a metabolic pathway act synergistically. It is found that two mutations tend to act mostly synergistically when they reduce the activity of the same enzyme. If the two mutations affect different enzymes, the conclusion depends on the way that fitness is determined by aspects of the pathway. The cases analyzed are: selection for (1) maximal flux, (2) maximal equilibrium concentration (pool size) of an intermediate, (3) optimal flux, (4) optimal pool size. The respective types of epistasis found are: (1) antagonistic, (2) partly synergistic, (3-4) synergism is likely to predominate over antagonism. This results in somewhat different predictions concerning the effect of metabolic mutations on fitness in prokaryotes and eukaryotes. The fact that bacteria are largely clonal but have often a mosaic gene structure is consistent with expectations from the model. PMID- 8417984 TI - Leber's hereditary optic neuropathy: the etiological role of a mutation in the mitochondrial cytochrome b gene. PMID- 8417985 TI - Lethal transposition of Mud phages in Rec- strains of Salmonella typhimurium. AB - Under several circumstances, the frequency with which Mud prophages form lysogens is apparently reduced in rec strains of Salmonella typhimurium. Lysogen formation by a MudI genome (37 kb) injected by a Mu virion is unaffected by a host rec mutation. However when the same MudI phage is injected by a phage P22 virion, lysogeny is reduced in a recA or recB mutant host. A host rec mutation reduces the lysogenization of mini-Mu phages injected by either Mu or P22 virions. When lysogen frequency is reduced by a host rec mutation, the surviving lysogens show an increased probability of carrying a deletion adjacent to the Mud insertion site. We propose that the rec effects seen are due to a failure of conservative Mu transposition. Replicative Mud transposition from a linear fragment causes a break in the host chromosome with a Mu prophage at both broken ends. These breaks are lethal unless repaired; repair can be achieved by Rec functions acting on the repeated Mu sequences or by secondary transposition events. In a normal Mu infection, the initial transposition from the injected fragment is conservative and does not break the chromosome. To account for the conditions under which rec effects are seen, we propose that conservative transposition of Mu depends on a protein that must be injected with the DNA. This protein can be injected by Mu but not by P22 virions. Injection or function of the protein may depend on its association with a particular Mu DNA sequence that is present and properly positioned in Mu capsids containing full-sized Mu or MudI genomes; this sequence may be lacking or abnormally positioned in the mini-Mud phages tested. PMID- 8417986 TI - Identification of Aspergillus brlA response elements (BREs) by genetic selection in yeast. AB - The brlA gene of Aspergillus nidulans plays a central role in controlling conidiophore development. To test the hypothesis that brlA encodes a transcriptional regulator and to identify sites of interaction for the BrlA polypeptide, we expressed brlA in Saccharomyces cerevisiae (yeast) strains containing Aspergillus DNA sequences inserted upstream of a minimal yeast promoter fused to the Escherichia coli lacZ gene. Initially, a DNA fragment from the promoter region of the developmentally regulated rodA gene was tested and shown to mediate brlA-dependent transcriptional activation. Two additional DNA fragments were selected from an Aspergillus genomic library by their ability to respond to brlA in yeast. These fragments contained multiple copies of a sequence motif present in the rodA fragment, which we propose to be sites for BrlA interaction and designate brlA response elements (BREs). DNA fragments containing BREs upstream of a minimal Aspergillus promoter were capable of conferring developmental regulation in Aspergillus. Deletion of BREs from the upstream region of rodA greatly decreased its developmental induction. Multiple copies of a synthetic oligonucleotide with the consensus sequence identified among the BREs mediated brlA-dependent transcriptional activation in yeast. The results show that a primary activity of brlA is transcriptional activation and tentatively identify sites of interaction for the BrlA polypeptide. PMID- 8417988 TI - Felix Bernstein and the first human marker locus. PMID- 8417987 TI - A Saccharomyces cerevisiae RAD52 allele expressing a C-terminal truncation protein: activities and intragenic complementation of missense mutations. AB - A nonsense allele of the yeast RAD52 gene, rad52-327, which expresses the N terminal 65% of the protein was compared to two missense alleles, rad52-1 and rad52-2, and to a deletion allele. While the rad52-1 and the deletion mutants have severe defects in DNA repair, recombination and sporulation, the rad52-327 and rad52-2 mutants retain either partial or complete capabilities in repair and recombination. These two mutants behave similarly in most tests of repair and recombination during mitotic growth. One difference between these two alleles is that a homozygous rad52-2 diploid fails to sporulate, whereas the homozygous rad52-327 diploid sporulates weakly. The low level of sporulation by the rad52 327 diploid is accompanied by a low percentage of spore viability. Among these viable spores the frequency of crossing over for markers along chromosome VII is the same as that found in wild-type spores. rad52-327 complements rad52-2 for repair and sporulation. Weaker intragenic complementation occurs between rad52 327 and rad52-1. PMID- 8417989 TI - Identification of new genes required for meiotic recombination in Saccharomyces cerevisiae. AB - Mutants defective in meiotic recombination were isolated from a disomic haploid strain of Saccharomyces cerevisiae by examining recombination within the leu2 and his4 heteroalleles located on chromosome III. The mutants were classified into two new complementation groups (MRE2 and MRE11) and eight previously identified groups, which include SPO11, HOP1, REC114, MRE4/MEK1 and genes in the RAD52 epistasis group. All of the mutants, in which the mutations in the new complementation groups are homozygous and diploid, can undergo premeiotic DNA synthesis and produce spores. The spores are, however, not viable. The mre2 and mre11 mutants produce viable spores in a spo13 background, in which meiosis I is bypassed, suggesting that these mutants are blocked at an early step in meiotic recombination. The mre2 mutant does not exhibit any unusual phenotype during mitosis and it is, thus, considered to have a mutation in a meiosis-specific gene. By contrast, the mre11 mutant is sensitive to damage to DNA by methyl methanesulfonate and exhibits a hyperrecombination phenotype in mitosis. Among six alleles of HOP1 that were isolated, an unusual pattern of intragenic complementation was observed. PMID- 8417990 TI - Identification of functionally related genes that stimulate early meiotic gene expression in yeast. AB - Meiosis and spore formation in the yeast Saccharomyces cerevisiae are associated with increased expression of sporulation-specific genes. One of these genes, IME2, encodes a putative protein kinase that is a positive regulator of other sporulation-specific genes. We have isolated mutations that cause reduced expression of an ime2-lacZ fusion gene. We found mutations in IME1, a known positive regultor of IME2, and MCK1, a known positive regulator of IME1. We also isolated recessive mutations in 12 other genes, which we designate RIM (Regulator of IME2) genes. Our analysis indicates that the defects in rim1, rim8, rim9 and rim13 mutants are a consequence of diminished IME1 expression and can be suppressed by expression of IME1 from the heterologous ACT1 promoter. These rim mutations also reduced expression of an ime1-HIS3 fusion, in which the HIS3 gene is expressed from the IME1 promoter, and caused reduced levels of IME1 RNA. Although the rim1, rim8, rim9 and rim13 mutant phenotypes are similar to those of mck1 mutants, we found that the defects in ime2-lacZ expression and sporulation of the mck1 rim double mutants were more severe than either single mutant. In contrast, the defects of the rim rim double mutants were similar to either single mutant. The rim1, rim8, rim9 and rim13 mutants also display slow growth at 17 degrees and share a smooth colony morphology that is not evident in mck1 mutants or isogenic wild-type strains. We suggest that RIM1, RIM8, RIM9 and RIM13 encode functionally related products that act in parallel to MCK1 to stimulate IME1 expression. PMID- 8417991 TI - The Caenorhabditis elegans spe-6 gene is required for major sperm protein assembly and shows second site non-complementation with an unlinked deficiency. AB - Caenorhabditis elegans spermatozoa move by crawling. Their motility requires thin cytoskeletal filaments assembled from a unique cytoskeletal protein, the major sperm protein (MSP). During normal sperm development the MSP is segregated to developing sperm by assembly into filaments that form a paracrystalline array in a transient organelle, the fibrous body-membranous organelle. Mutations in the spe-6 gene cause sterility because they lead to defective primary spermatocytes that do not form spermatids. In these mutant spermatocytes the MSP fails to assemble into fibrous body filaments. Instead, the unassembled MSP distributes throughout the cytoplasm and nucleus. Thus, the spe-6 gene product is necessary for normal MSP localization and assembly during sperm development. In addition to their MSP assembly defect, spe-6 mutant spermatocytes arrest meiosis at diakinesis although their spindle pole bodies still replicate and separate. This results in spermatocytes with four half-spindles surrounding condensed, but unsegregated, chromosomes. All four spe-6 alleles, as well as a chromosome III deficiency that deletes the spe-6 gene, fail to complement two small overlapping chromosome IV deficiencies, eDf18 and eDf19. This non-allele-specific second site non-complementation suggests a concentration-dependent interaction between the spe-6 gene product and products of the gene(s) under eDf18 and eDf19, which include a cluster of sperm-specific genes. Since MSP filament assembly is highly concentration-dependent in vitro, the non-complementation might be expected if the sperm-specific gene products under eDf18 and eDf19 were needed together with the spe-6 gene product to promote MSP assembly. PMID- 8417994 TI - Hospitals work through superfund citations. PMID- 8417992 TI - Characterization of the mus308 gene in Drosophila melanogaster. AB - Among the available mutagen-sensitive mutations in Drosophila, those at the mus308 locus are unique in conferring hypersensitivity to DNA cross-linking agents but not to monofunctional agents. Those mutations are also associated with an elevated frequency of chromosomal aberrations, altered DNA metabolism and the modification of a deoxyribonuclease. This spectrum of phenotypes is shared with selected mammalian mutations including Fanconi anemia in humans. In anticipation of the molecular characterization of the mus308 gene, it has been localized cytogenetically to 87C9-87D1,2 on the right arm of chromosome three. Nine new mutant alleles of the gene have been generated by X-ray mutagenesis and one was recovered following hybrid dysgenesis. Characterization of these new alleles has uncovered additional phenotypes of mutations at this locus. Homozygous mus308 flies that have survived moderate mutagen treatment exhibit an altered wing position that is correlated with reduced flight ability and an altered mitochondrial morphology. In addition, observations of elevated embryo mortality are potentially explained by an aberrant distribution of nuclear material in early embryos which is similar to that seen in the mutant giant nuclei. PMID- 8417993 TI - The mitochondrial genome of the honeybee Apis mellifera: complete sequence and genome organization. AB - The complete sequence of honeybee (Apis mellifera) mitochondrial DNA is reported being 16,343 bp long in the strain sequenced. Relative to their positions in the Drosophila map, 11 of the tRNA genes are in altered positions, but the other genes and regions are in the same relative positions. Comparisons of the predicted protein sequences indicate that the honeybee mitochondrial genetic code is the same as that for Drosophila; but the anticodons of two tRNAs differ between these two insects. The base composition shows extreme bias, being 84.9% AT (cf. 78.6% in Drosophila yakuba). In protein-encoding genes, the AT bias is strongest at the third codon positions (which in some cases lack guanines altogether), and least in second codon positions. Multiple stepwise regression analysis of the predicted products of the protein-encoding genes shows a significant association between the numbers of occurrences of amino acids and %T in codon family, but not with the number of codons per codon family or other parameters associated with codon family base composition. Differences in amino acid abundances are apparent between the predicted Apis and Drosophila proteins, with a relative abundance in the Apis proteins of lysine and a relative deficiency of alanine. Drosophila alanine residues are as often replaced by serine as conserved in Apis. The differences in abundances between Drosophila and Apis are associated with %AT in the codon families, and the degree of divergence in amino acid composition between proteins correlates with the divergence in %AT at the second codon positions. Overall, transversions are about twice as abundant as transitions when comparing Drosophila and Apis protein-encoding genes, but this ratio varies between codon positions. Marked excesses of transitions over chance expectation are seen for the third positions of protein-coding genes and for the gene for the small subunit of ribosomal RNA. For the third codon positions the excess of transitions is adequately explained as due to the restriction of observable substitutions to transitions for conserved amino acids with two-codon families; the excess of transitions over expectation for the small ribosomal subunit suggests that the conservation of nucleotide size is favored by selection. PMID- 8417995 TI - A greener image. Hospitals take on environmental challenges of the '90s. PMID- 8417996 TI - Alta Bates Medical Center. Proper waste sorting begins with good definitions. PMID- 8417997 TI - What organized labor wants. The principles are the same, but labor is more flexible. PMID- 8417998 TI - Clinical quandaries. Getting MDs to buy into CQI means making adjustments. PMID- 8417999 TI - Geriatric services grow. Hospitals work together to fill gaps in continuum of care. PMID- 8418000 TI - What the upper midwest knows. Region leads the nation in net patient revenues. PMID- 8418001 TI - Toppling the power of the pyramid. Team-based restructuring for TQM, patient centered care. PMID- 8418002 TI - 'UR' pharmacists help MDs control drug costs. PMID- 8418003 TI - Hospitals increase med staff use of IS. PMID- 8418004 TI - Social workers in the ED tackle many problems. PMID- 8418005 TI - AHCPR chief cites achievements after three years. Interview by Mark M. Hagland. PMID- 8418006 TI - Experts: hospitals overdoing sterile supply procedure. PMID- 8418007 TI - Data watch. Survey outlines hospital employee health benefits. PMID- 8418008 TI - Don't let uncertainty keep you from action. PMID- 8418009 TI - Structural and functional aspects of transforming growth factor-beta in prostate cancer and other human malignancies. PMID- 8418010 TI - Prolactin presents in all pituitary tumors of acromegalic patients. AB - Twenty-two consecutive cases of adenoma in acromegalic patients were studied immunohistochemically. All the tumors contained prolactin (PRL)-reactive cells (3% to 53% of the total number of tumor cells) as well as growth hormone (GH) reactive cells (4% to 74% of the total number of tumor cells). All acromegalic cases studied were thus plurihormonal adenomas containing GH and PRL; no pure GH cell adenoma was present. Twenty cases were further examined at the ultrastructural level in conjunction with postembedding double-labeling immunoelectron microscopy; 15 of these cases were diagnosed as mixed GH cell-PRL cell adenomas. The previously diagnosed pure GH cell adenomas possibly may have contained PRL cells and thus should be considered as mixed GH cell-PRL cell adenomas. Mammosomatotroph adenomas were rare in this series. Double-labeling immunoelectron microscopy, using protein A gold particles of two different sizes, greatly facilitated the distinction among GH, PRL, and mammosomatotroph cells. PMID- 8418011 TI - Rabies encephalitis in a patient with no history of exposure. AB - A 29-year-old man died of a rapidly progressive encephalitis without a clinical diagnosis and without a history of exposure to the rabies virus. A diagnosis of rabies was established postmortem by histologic and ultrastructural demonstration of rabies virus inclusions, by fluorescent antibody reaction, and by viral culture. Viral inclusions were sparse, were generally irregular and poorly demarcated, and were confined almost exclusively to the cerebellar Purkinje cells. A history of exposure was obtained only in retrospect on detailed questioning of friends and relatives. Rabies encephalitis is now seen most frequently in patients without a history of exposure and may be easily overlooked both clinically and pathologically. PMID- 8418012 TI - Classification of gastric carcinomas. PMID- 8418013 TI - Immunohistologic studies of bone marrow biopsies on frozen sections: an analysis of 42 cases. AB - An immunohistologic study of bone marrow biopsy frozen sections from 42 cases involved by a variety of reactive and neoplastic disorders is presented. Thirteen cases also were studied using other methods, including cytochemistry, surface marker analysis of cell suspensions, and/or DNA hybridization. Thirty-four of 42 cases (81%) were adequately phenotyped on frozen tissue using a panel of antibodies for hematolymphoid-associated antigens. The immunostains from the remaining eight cases were unsatisfactory, primarily as a result of heavy background staining. Eighteen cases were lymphoproliferative disorders of B-cell phenotype and 12 of these showed surface monotypic immunoglobulin expression by the frozen section technique. Six cases showed B- or pre-B-cell antigens but no surface immunoglobulins. Of the remaining 16 patients, two cases showed myeloid markers and three showed T-cell phenotype. Nine cases showed a mixture of polyclonal B- and T-cell populations. Keratin was demonstrated in a single case of metastatic carcinoma included in the study. These results indicate that the majority of hematopoietic processes can be successfully phenotyped on bone core frozen sections and demonstrate the usefulness of immunohistologic study of the frozen bone marrow biopsy specimens, especially when the specimens for other modalities are not available or are inadequate. The keys to achieving the best results from the frozen bone marrow immunohistochemistry were the gentle handling of the specimens and the preparation of high-quality, cryostat-cut frozen sections. PMID- 8418014 TI - Diagnostic considerations in molar gestations. AB - Hydatidiform moles (HMs) are classified as partial or complete based on a combination of gross, histologic, and karyotypic features. Adherence to strict and reproducible diagnostic criteria is needed to ensure accurate diagnosis and minimize interpathologist variability. Using the kappa statistic as a measure of agreement, the morphologic, flow cytometric, and clinical features of 80 cases of HM or suspected HM were analyzed sequentially by three pathologists to evaluate intrapathologist and interpathologist variability. Poor interpathologist agreement was obtained when histology alone was used for diagnosis. The combination of gross morphology and histology resulted in poor to good agreement. Good interpathologist agreement was obtained, however, when objective data (DNA content determined by flow cytometry) were included in the analysis. Our data indicate that pathologist concordance is maximized when the diagnosis is based on a combination of morphology and DNA content. PMID- 8418015 TI - The nonlymphoid microenvironment of reactive follicles and lymphomas of follicular origin as defined by immunohistology on paraffin-embedded tissues. AB - Twenty-five reactive lymph nodes, 10 palatine tonsils, and 72 B-cell non Hodgkin's lymphomas (NHLs) of supposed follicular origin were investigated in an immunohistologic study of fixed, paraffin-embedded tissues using a panel of monoclonal antibodies reactive with antigens resistant against fixation and paraffin-embedding techniques together with polyclonal antibodies. The results concerning the microenvironmental organization of reactive lymphoid follicles confirmed that the distribution of CD21+ and CD23+ dendritic reticulum cells, vimentin+ fibroblastic reticulum cells, and CD68+ tingible-body macrophages is heterogeneous with reference to their immunostaining patterns and topographic localization within the germinal center and mantle zone. Moreover, a close microenvironmental similarity between the follicular lymphomas of supposed germinal center or mantle zone origin and their normal counterparts was noted. The study of the microenvironment of the B-zone small lymphocytic lymphoma cases, showing the same distribution patterns for the nonlymphoid cells as seen in mantle zone lymphomas, corroborated the supposed follicular origin of this unusual variant of small lymphocytic lymphoma. In conclusion, this study shows that monoclonal antibodies recognizing CD21, CD23, and CD68 antigens may be valuable additions to vimentin, S-100 protein, laminin, and type IV collagen antibodies for investigating the microenvironmental organization of lymphoid tissues in both normal and neoplastic conditions. PMID- 8418016 TI - Focal mesangiolysis and the pathogenesis of the Kimmelstiel-Wilson nodule. AB - Kidneys from 74 consecutively autopsied primarily non-insulin-dependent diabetes cases and 59 age-, sex-, and ethnic group-matched controls were examined qualitatively and semiquantitatively to determine whether focal mesangiolyses (FMs), Kimmelstiel-Wilson (KW) nodules, and glomerular capillary microaneurysms (GCMs) were related lesions, to determine their extent and pathogenic sequence, and to look for associations with structural and functional factors. Light microscopic examination of serial sections, immunohistochemical stains, image analysis, and electron microscopy were used. Focal mesangiolyses, KW nodules, and GCMs occurred in 31 of the 74 diabetes cases (27 had FMs, 29 had KW nodules, and nine had GCMs) and were positively correlated with each other semiquantitatively (r = .71, .70, and .68, respectively). Numerous FMs were found, involving 62% and 78% of the glomeruli in the two most severely affected cases. Most FMs were located at the periphery of KW nodules, but de novo FMs were documented in six cases. Glomerular capillary microaneurysms were deemed occasional complications of FMs because they were much less common, and 25 of the 27 GCMs identified were contiguous with FMs. Focal mesangiolyses and GCMs were deemed transient lesions, being absent in end-stage kidneys. Both FMs and KW nodules consisted of a spectrum of lesions. For the sake of clarity they were arbitrarily divided into two types: edematous and proliferative FMs and simple and complicated KW nodules. Their characteristics suggested the following pathogenic sequence: edematous FM- >proliferative FM-->focal nodular mesangial expansion-->simple KW nodule- >recurrent FM-->complicated KW nodule. Complicated nodules were associated with marked alterations in the lobular capillary. The number of mesangial cells was increased in FMs and they were thought to be responsible for increased matrix production. Focal mesangiolyses and KW nodules were positively associated with diabetes, proteinuria, and hyalinization of afferent and efferent arterioles, but were weakly or not associated with hypertension, arcuate and interlobular artery stenosis, hydroenphrosis, acute pyelonephritis, renal arterial atheromatous emboli, glomerular platelet-fibrin thromboemboli, and congestive heart failure. PMID- 8418017 TI - An immunohistochemical evaluation of androgen and progesterone receptors in ovarian tumors. AB - To visualize the heterogeneity of androgen receptor (AR) and progesterone receptor (PR) expression in ovarian tumors, 61 specimens were studied immunohistochemically using specific mouse monoclonal antibodies. The tested ovarian tumors included ovarian carcinomas of serous, mucinous, endometrioid, undifferentiated, and clear cell types as well as borderline epithelial tumors, mucinous cystadenomas, granulosa cell tumors, metastatic ovarian tumors, and one case of immature teratoma. Sixty-seven percent of ovarian carcinomas expressed AR, while 46% of the ovarian carcinomas expressed PR. In addition, the three tested borderline epithelial tumors were AR positive. Androgen receptor and PR were primarily localized in the nuclei of ovarian tumor cells. In three tumors focal staining of intervening stroma cells was observed. Interestingly, 35 of the granulosa cell tumors expressed PR as well as AR. All tumor types displayed a wide range in their proportions of AR- and PR-positive tumor cells. A poor correlation, however, was observed when semiquantitative immunohistochemical AR staining results on 17 ovarian carcinomas were compared with biochemical cytosol AR estimations using the dextran-coated charcoal method. Our study indicates that AR as well as PR is expressed by a substantial proportion of ovarian borderline and malignant tumors and that there is a strong heterogeneity in expression of AR and PR. It is possible to evaluate the prognostic significance of these receptors in ovarian cancers on fresh-frozen tumor specimens using a simple immunohistochemical technique. PMID- 8418018 TI - Smaller local brain volumes and cerebral atrophy in spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) have enlarged cerebral ventricles from 8 weeks of age onward and smaller brains than age-matched, normotensive Wistar Kyoto (WKY) rats (controls). At 6-7 months of age local cerebral glucose utilization is apparently lower in many brain areas of SHR relative to WKY rats. These observations led to the hypothesis that there are morphological differences between these two strains of rats in many, if not all, brain areas. This hypothesis was tested in 6-7-month-old SHR and WKY rats by quantitating 1) the volumes of the ventricular system, whole brain, six gray matter structures, and two white matter areas; 2) the thickness of two regions of the cerebral cortex; and 3) the frequency of neuronal nuclei (neuronal frequency) in nine brain areas. Ventricular volume was twofold greater in SHR than in control rats. The volumes of the entire brain and all six gray matter structures plus the thickness of the two cortical regions were 11-25% less in SHR. Neuronal frequency was, however, similar in the two rat strains. The latter finding coupled with the smaller regional tissue volumes indicates appreciably fewer neurons per brain structure in young adult SHR than in controls. These results indicate significant cerebral structural differences between young adult SHR and WKY rats and suggest that structure as well as metabolism are abnormal in the SHR brain. PMID- 8418019 TI - Calcium dependence of flow-induced dilation. Cooperative interaction with sodium. AB - The effect of changing extracellular calcium and sodium concentrations on flow, acetylcholine, and papaverine vasodilation and also on norepinephrine contraction was studied in a segment of a resistance branch of the rabbit central ear artery mounted in a myograph. Decreases in calcium to 80% of the normal physiological saline solution concentration (1.6 mM) reduced both flow- and acetylcholine induced dilation. Increases of calcium to 120%, 140%, and 200% of normal decreased flow dilation responses, but not those to acetylcholine and papaverine. Thus, the optimum calcium concentration for flow dilation lies within the range of 1.4-1.9 mM. The concomitant proportionate reduction of sodium and calcium offsets the reduction in flow dilation that occurred with reduction in calcium alone. This was true whether sodium and calcium were reduced simultaneously or whether the effect of lowered sodium and then that of lowered sodium and calcium was studied. Emphasizing the uniqueness of this interaction between sodium and calcium are the observations that the depression of acetylcholine dilation by calcium reduction was not influenced by a concurrent reduction in sodium and that the depression of flow dilation caused by sodium reduction is increased by calcium increase, which by itself depresses flow dilation. None of these changes in sodium and calcium alters the responses of the artery segment to papaverine or norepinephrine. We propose that these interactions of sodium and calcium in relation to flow dilation may reflect the binding properties for sodium and calcium of a proposed flow sensor, the glycosaminoglycan polyanions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418020 TI - Marine oils dose-dependently inhibit vasoconstriction of forearm resistance vessels in humans. AB - The effects of dietary supplementation with marine oils on vascular reactivity in human forearm resistance arteries were studied. Healthy male adults (six to nine subjects per group) were given either maxEPA capsules (content: eicosapentaenoic acid, 0.178 g/g; docosahexaenoic acid, 0.116 g/g) at doses of 20, 10, or 5 g/day or placebo capsules at 20 g/day for 28 days. Capsule compliance was confirmed by measurement of platelet membrane incorporation of n-3 fatty acids. Blood pressure was not affected by either maxEPA or placebo. The influence of treatment interventions on forearm vasoconstrictive responses to local infusions of angiotensin II and norepinephrine was examined using venous occlusion plethysmography before and after treatment. Responses to both agonists were significantly suppressed by 20 g/day maxEPA (slopes before and after maxEPA, respectively: angiotensin II, 3.34 and 0.89; norepinephrine, 0.91 and 0.41). When analyzed as difference in area under the dose-response curves, the suppressive effects of maxEPA were clearly dose dependent (angiotensin II: 20 g area reduced by 72%, 10 g by 67%, 5 g by 33%). Similarly, responses to norepinephrine were dose-dependently suppressed by maxEPA (20 g area reduced by 61%, 10 g by 63%, and 5 g by 33%). Placebo had no effect on the responses to either constrictor. The responses to both agonists returned to preoil levels after 2 months' discontinuation of 20 g/day maxEPA. We conclude that the suppressive effects of marine oils on vascular reactivity may, in part, contribute to their cardioprotective influence in humans. PMID- 8418021 TI - Angiotensin II causes mesangial cell hypertrophy. AB - Angiotensin II, a potent vasoconstrictor and known growth factor for vascular smooth muscle cells, has been implicated in the development of glomerulosclerosis. Because mesangial cell growth plays a critical role in the glomerulosclerotic process, the objective of this study was to determine the direct effect of long-term (48-hour) angiotensin II treatment on the growth of cultured murine mesangial cells. Subconfluent, quiescent adult murine mesangial cells were treated for 48 hours with media containing angiotensin II with and without its specific inhibitor losartan. In comparison to cells treated with serum-free medium, cells treated with serum plus insulin demonstrated a significant increase in cell number (1.93 +/- 0.1 times control, p < 0.05), [3H]thymidine incorporation per 10(5) cells (2.29 +/- 0.12 times control, p < 0.05), [3H]leucine incorporation per 10(5) cells (1.81 +/- 0.18 times control, p < 0.05), and total protein content per 10(5) cells (1.65 +/- 0.07 times control, p < 0.05). In contrast, cells treated with angiotensin II (10(-6) M) had no significant increase in cell number (0.84 +/- 0.01 times control) or [3H]thymidine incorporation per 10(5) cells (1.23 +/- 0.12 times control) but demonstrated a significant increase in [3H]leucine incorporation per 10(5) cells (1.61 +/- 0.09 times control) and total protein content per 10(5) cells (1.38 +/- 0.04 times control). Pretreatment with losartan blocked 56% of the angiotensin II induced increase in [3H]leucine incorporation and 84% of the angiotensin II induced increase in total protein content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418022 TI - Time course of stimulation of renal renin messenger RNA by furosemide. AB - Renin secretion responds rapidly to a variety of stimuli; however, reported changes in renal renin messenger RNA (mRNA) levels in vivo have been observed only after prolonged stimulation. Studies were designed to test whether rapid changes in renin mRNA levels can be produced in vivo. In the first series, Sprague-Dawley rats received furosemide (10 mg/kg) intraperitoneally and a low sodium diet (0.05% sodium); renin secretion was significantly stimulated at 8 or 16 hours after treatment, but renin mRNA levels did not change. In a second series, rats were pretreated with deoxycorticosterone acetate (200 mg/kg) and saline drinking water for 3 days and then killed 0, 2, 4, 8, or 48 hours after furosemide administration. The renin mRNA level was unchanged at 2 hours but was stimulated twofold at 4 and 8 hours and threefold at 48 hours. In additional animals, the response of renin mRNA 4 hours after furosemide was found not to be potentiated by the converting enzyme inhibitor quinapril (5 mg/kg). The results demonstrate that with acute stimulation, renin mRNA levels lag 2-4 hours behind the change in plasma renin levels. PMID- 8418023 TI - Angiotensin II, sodium, and cardiovascular hypertrophy in spontaneously hypertensive rats. AB - Angiotensin II (Ang II) may cause cardiovascular hypertrophy as a consequence of increased blood pressure or possibly by direct trophic actions. To dissociate Ang II and blood pressure in young spontaneously hypertensive rats (SHR), we used sodium loading during angiotensin converting enzyme inhibitor treatment. Animals were treated between 6 and 10 weeks of age with perindopril to lower Ang II and blood pressure, or with perindopril and 1% saline drinking fluid or perindopril and aldosterone infusion to lower Ang II but maintain high blood pressure. Blood pressure, heart weight, and media/lumen ratio of mesenteric resistance arteries were studied while rats were on treatment at 10 weeks of age and 15 weeks after treatment at 25 weeks of age. Perindopril lowered blood pressure and inhibited the development of cardiovascular hypertrophy. Saline or aldosterone restored high blood pressure during perindopril treatment and resulted in increased heart weight/body weight and resistance artery media/lumen ratios in direct proportion to the elevation of blood pressure. Because increased structure occurred despite perindopril treatment, we conclude that direct trophic actions of Ang II are not essential for the development of cardiovascular hypertrophy in young SHR and that the antitrophic actions of angiotensin converting enzyme inhibitors depend more on changes in blood pressure than on Ang II. However, restoration of blood pressure and structure by sodium during perindopril treatment raises the possibility that the design of the cardiovascular system and blood pressure may depend indirectly on Ang II through effects on sodium metabolism. PMID- 8418024 TI - Oxygen cost of stress development in hypertrophied and failing hearts from the spontaneously hypertensive rat. AB - Left ventricular isovolumic stress development and metabolic parameters were studied in 18-24-month-old spontaneously hypertensive rats (SHRs) and age-matched Wistar-Kyoto (WKY) rat controls using the isolated, isovolumic (balloon in left ventricle) buffer-perfused rat heart preparation. After WKY rats and all SHRs were compared, SHRs were divided into two groups: those animals with (SHR-F) and without (SHR-NF) evidence of heart failure. Hearts were perfused at 100 mm Hg using a constant pressure system at a temperature of 37 degrees C. In the baseline state, peak systolic pressure was greatest in the SHR-NF group and lowest in the SHR-F group. Peak midwall stress was greatest in the WKY group and, again, lowest in the SHR-F group. Oxygen consumption was lowest in the SHR-F group. When the oxygen cost of stress development was estimated by normalizing myocardial oxygen consumption by peak developed midwall stress, values were lowest in the WKY, greater in the SHR-NF, and greatest in the SHR-F group. Lactate production did not occur in the baseline state in any of the groups. Functional and metabolic responses to graded hypoxia, induced by changing the gas mixture of the perfusate from 95% to 50%, 25%, and 0% oxygen at perfusion pressures of 100 and 130 mm Hg, were studied. Increasing perfusion pressure generally resulted in small increases in peak systolic pressure and myocardial oxygen consumption but did not substantially reverse the contractile or metabolic deficit present in the SHR-F group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418025 TI - Feasibility of ambulatory, continuous 24-hour finger arterial pressure recording. AB - We tested Portapres, an innovative portable, battery-operated device for the continuous, noninvasive, 24-hour ambulatory measurement of blood pressure in the finger. Portapres is based on Finapres, a stationary device for the measurement of finger arterial pressure. Systems were added to record signals on tape, to alternate measurements between fingers automatically each 30 minutes, and to correct for the hydrostatic height of the hand. We compared the pressure as measured by Portapres with contralateral intrabrachial pressure measured with an Oxford device. Results were obtained in eight volunteers and 16 hypertensive patients. Time lost due to artifact was about 10% for each device. In two patients a full 24-hour Oxford profile was not obtained. In the remaining 22 subjects finger systolic, diastolic, and mean pressures differed +1 (SD 9), -8 (6), and -10 (6) mm Hg, respectively, from intrabrachial pressure. These diastolic and mean pressure underestimations are similar to what was found earlier for Finapres, are typical for the technique, and are systematic. Avoiding brisk hand movements resulted in fewer waveform artifacts. The hand had to be kept covered to continue recording at low outside temperatures. Sleep was not disturbed by Portapres, and arterial pressure showed a marked fall during siesta and nighttime. There were no major limitations in behavior, and no discomfort that originated from continuous monitoring was reported. Measurements continued normally during physical exercise. Portapres provides for the first time continuous 24-hour, noninvasive ambulatory blood pressure waveform monitoring and offers real and obvious advantages over current noninvasive and invasive devices. PMID- 8418026 TI - Validation of a method for noninvasive measurement of central arterial pressure. AB - The goal of this study was to validate a newly improved noninvasive method for calibrated measurement of the ascending portion of the central arterial pressure wave in humans. Noninvasive pressure waveforms were generated by measuring the time delay between the R wave of the electrocardiogram and onset of brachial artery flow (by Doppler) during computer-controlled upper arm cuff deflation. This delay shortens with falling cuff pressure (becoming near constant at and below diastolic pressure), so that a plot of pressure versus time delay yields the ascending portion of the arterial waveform. These waveforms were compared with simultaneous invasive ascending aortic pressures in 57 adult patients (31 by fluid manometer [group A] and 26 by catheter-tipped micromanometer [group B]) during routine cardiac catheterization. Patient age ranged from 26 to 77 years. Eighty percent of group A patients and 40% of group B had coronary artery disease. Noninvasive systolic and diastolic pressures were very similar to invasive values in both groups (Pni = 0.98 x Pi, r = 0.99, p < 0.0001). Instantaneous pressure differences between waveforms were also similar in both groups, averaging between 4.5 and 5.5 mm Hg. Micromanometer and noninvasive pressure data were also obtained before and after intravenous nitroglycerin (n = 5) and isometric handgrip (n = 8) and demonstrated good agreement. A potential application of these pressures is for estimating maximal ventricular power to assess systolic function. This was tested using invasive pressure-volume data from four patients under a variety of conditions (exercise, pacing, etc.).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418027 TI - Level of control of hypertension in Mexican Americans and non-Hispanic whites. AB - Compared with non-Hispanic whites, Mexican Americans have a higher prevalence of diabetes, greater adiposity, and an unfavorable body fat distribution. The prevalence of hypertension, however, is similar or lower in Mexican Americans than in non-Hispanic whites. There is little information on the level of blood pressure control in Mexican Americans. We compared the mean blood pressure levels of Mexican American and non-Hispanic white hypertensive subjects in the San Antonio Heart Study, a population-based study of diabetes and cardiovascular disease. Hypertension was defined as one or more of a systolic blood pressure > or = 160 mm Hg, a diastolic blood pressure > or = 95 mm Hg, and current use of antihypertensive medications. Three hundred and fifty-eight Mexican Americans and 241 non-Hispanic whites met these criteria. Poor hypertension control was defined as a systolic blood pressure > or = 160, a diastolic blood pressure > or = 95 mm Hg, or both. After adjustment for age, gender, obesity, body fat distribution, and level of educational attainment, Mexican American hypertensive subjects were in significantly poorer control than non-Hispanic white hypertensive subjects. The reasons for their poorer control are unknown, but our findings emphasize the importance of hypertension in this ethnic group. PMID- 8418028 TI - Restenosis after a first percutaneous transluminal renal angioplasty. AB - The incidence of restenosis after a first successful percutaneous transluminal angioplasty of a native renal artery and the clinical and angiographic variables that may influence its occurrence were studied in 104 hypertensive patients. Angiograms obtained immediately before and after angioplasty and, in 92 patients, 8.8 +/- 6.0 months after angioplasty were interpreted separately by two observers. Stenosis severity was classified into five grades, and restenosis was defined by a stenosis one grade or more higher at follow-up than immediately after angioplasty. Interobserver concordance for etiology, stenosis grade, and other angiographic items yielded kappa coefficients in the range of 0.328-0.942. Sessions were organized to reach a consensus in each case. Ostial stenoses were more frequent in patients with atheromatous stenoses, and branch stenoses were more frequent in those with fibromuscular dysplasia. There was no significant difference between the 15 patients (16%) with restenosis and those without concerning sex distribution, mean age, mean blood pressure, plasma creatinine level, and etiology distribution. Truncal stenoses were less prone to restenosis than ostial or branch stenoses (12% versus 35%, respectively; 95% confidence interval of difference, -0.6% to 47%). In patients with atheromatous stenoses, aortitis or aortic ectasia were associated with a high restenosis incidence (35% when present versus 8% when absent; 95% confidence interval of difference, 5% to 48%). In conclusion, restenosis was observed in one sixth of patients after a first successful renal angioplasty; its incidence was low in patients with truncal stenoses and high in those with severe aortic atheroma. Automated renal artery stenosis quantification methods are needed to standardize stenosis description. PMID- 8418029 TI - Role of endothelium in endothelin-evoked contractions in the rat aorta. AB - We designed experiments to determine the role of endothelium-derived contracting factor or factors in the response to endothelin-1 and endothelin-3 in the aorta of normotensive and hypertensive rats. Rings of thoracic aortas, with and without endothelium, from normotensive and spontaneously hypertensive rats were suspended in organ chambers for recording of isometric tension in the presence of nitro-L arginine, an inhibitor of nitric oxide synthase. The removal of endothelium decreased the contractions evoked by both endothelins in the aorta of spontaneously hypertensive but not of normotensive rats. Indomethacin (inhibitor of cyclooxygenase), dazoxiben (inhibitor of thromboxane synthase), and SQ-29,548 (antagonist of thromboxane A2 receptors) reduced, in aortic rings of spontaneously hypertensive rats, the contractions to endothelins in rings with but not in those without endothelium, whereas their effect was not endothelium dependent in tissues of normotensive rats. BQ-123, a selective endothelin-A receptor antagonist, shifted the concentration-response curve to endothelin-1 to the right in a concentration-dependent manner and abolished the endothelium dependent component of the contractions evoked by the peptide. The presence of the endothelium increased the basal and endothelin-stimulated release of thromboxane B2, the stable metabolite of thromboxane A2, in aortas of spontaneously hypertensive rats but not in those of normotensive rats. These data suggest that endothelium-derived thromboxane A2 contributes to contractions evoked by endothelin-1 and endothelin-3 in the aorta of the spontaneously hypertensive rat but not in that of the normotensive rat. Both the receptors on the endothelial cells (mediating the release of thromboxane A2) and those on vascular smooth muscle belong to the endothelin-A subtype. PMID- 8418030 TI - Doxazosin and captopril in mildly hypercholesterolemic hypertensive patients. The Doxazosin-Captopril in Hypercholesterolemic Hypertensives Study. AB - The evidence linking hypertension and hypercholesterolemia is strong and has fueled research into possible adverse effects of some antihypertensive agents on serum lipid profile. This multicenter, open, parallel study compares the effects of doxazosin and captopril on blood pressure, serum lipid levels, and quality of life in 224 hypercholesterolemic hypertensive patients. Blood pressure was significantly reduced in both treatment groups (p < 0.001) and was normalized (standing diastolic pressure < or = 90 mm Hg) in 73% of the doxazosin patients and 67% of the captopril group. Serum total cholesterol level was favorably reduced by both doxazosin (from 238 to 223 mg/dl, p < 0.001) and captopril (from 245 to 233 mg/dl, p < 0.001), whereas high density lipoprotein cholesterol concentration increased only in the doxazosin group (from 33 to 36 mg/dl, p < 0.001). The calculated 10-year risk for the development of coronary heart disease was reduced significantly (p < 0.001) by 28% in the doxazosin group and by 19% in the captopril group. The quality of life evaluation showed beneficial changes in both treatment groups. As a result of proven antihypertensive efficacy and a lack of unfavorable effects on lipid parameters and health status measures, these findings support the use of both doxazosin and captopril as agents of first choice in the treatment of hypertensive patients with associated lipid abnormalities. PMID- 8418032 TI - Immunoglobulin M and G antibody responses to Plasmodium falciparum glutamate-rich protein: correlation with clinical immunity in Gambian children. AB - The aims of the present study were to describe the age-related immunoglobulin M (IgM) and IgG response to part of a 220-kDa glutamate-rich protein (GLURP) from Plasmodium falciparum and to determine possible correlations of possession of these antibodies with malaria morbidity. IgM and IgG levels were measured with a recombinant fusion protein consisting of the carboxy-terminal 783 amino acids of the GLURP. Samples for the study were obtained during a longitudinal malaria morbidity survey performed in The Gambia; cross-sectional surveys were performed at the beginning of the transmission season in May and in October. Seropositivity rates increased with age to a maximum of 77% for IgM and 95% for IgG in adults. High prevalences of seropositivity were associated with certain human leukocyte antigen class II alleles (DRw8, DR9, DR7, DR4, DQw7, and DQw2) or haplotypes. The relationship between anti-GLURP489-1271 antibodies and clinical immunity is not clear; asymptomatically infected children aged 5 to 8 years had significantly higher levels of IgG than clinically ill children of the same age, suggesting that antibodies to the carboxy-terminal part of the GLURP may contribute to immunity to P. falciparum. However, this was not significant for younger children. PMID- 8418031 TI - Isolation and partial characterization of glycolipid fractions from Mycobacterium avium serovar 2 (Mycobacterium paratuberculosis 18) that inhibit activated macrophages. AB - Glycolipid fractions from Mycobacterium avium serovar 2 (Mycobacterium paratuberculosis 18) inhibited the killing of Candida albicans by activated bovine peripheral-blood-derived macrophages. Fractions were derived by using the matrix solid-phase dispersion technique, which is a new method of simultaneous lysis and partial fractionation of components of bacterial cells. Further purification of active fractions was performed by concanavalin A affinity chromatography, centrifugal filtration, and differing solvent solubility. Three different fractions were isolated and partially characterized. Two of these fractions have characteristics typical of glycolipids, and the third fraction has characteristics compatible with a peptidoglycolipid. This peptidoglycolipid fraction has been purified and named MIF-A3. PMID- 8418033 TI - Complete nucleotide sequence of the Actinomyces viscosus T14V sialidase gene: presence of a conserved repeating sequence among strains of Actinomyces spp. AB - The nucleotide sequence of the Actinomyces viscosus T14V sialidase gene (nanH) and flanking regions was determined. An open reading frame of 2,703 nucleotides that encodes a predominately hydrophobic protein of 901 amino acids (M(r), 92,871) was identified. The amino acid sequence at the amino terminus of the predicted protein exhibited properties characteristic of a typical leader peptide. Five 12-amino-acid units that shared between 33 and 67% sequence identity were noted within the central domain of the protein. Each unit contained the sequence Ser-X-Asp-X-Gly-X-Thr-Trp, which is conserved among other bacterial and trypanosoma sp. sialidases. Thus, the A. viscosus T14V nanH gene and the other prokaryotic and eukaryotic sialidase genes evolved from a common ancestor. Southern hybridization analyses under conditions of high stringency revealed the existence of DNA sequences homologous to A. viscosus T14V nanH in the genomes of 18 strains of five Actinomyces species that expressed various levels of sialidase activity. The data demonstrate that the sialidase genes from divergent groups of Actinomyces spp. are highly conserved. PMID- 8418034 TI - Effects of cytokines on intracellular growth of Brucella abortus. AB - Interleukin 1 alpha (IL-1 alpha), IL-2, IL-4, IL-6, gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), and granulocyte macrophage colony stimulating factor (GM-CSF) were tested for their abilities to alter the growth of Brucella abortus in BALB/c J774A.1 murine macrophages. IL-1 alpha, IL-4, IL-6, tumor necrosis factor alpha, and granulocyte macrophage-colony-stimulating factor had no consistent or significant effect on the growth of the avirulent B. abortus strain 19. In contrast, the addition of either IFN-gamma or IL-2 at 100 U/ml to the macrophage cultures resulted in a significant reduction in the number of intracellular bacteria that was not attributable to decreased infection rates. With IL-2, the reduction was most often apparent only during the first 24 h after infection, while inhibition with IFN-gamma was apparent throughout the culture period of 48 h. The addition of either IL-2 or IFN-gamma to macrophage cultures also resulted in reduced intracellular CFU of the virulent B. abortus strain 2308 and the attenuated rough mutant B. abortus strain RB51. Inhibition of intracellular growth was not augmented by combinations of cytokines. Additional studies with IFN-gamma and IL-2 indicated that they could mediate the inhibition of intracellular growth of B. abortus in resident and thioglycolate broth-induced BALB/c peritoneal macrophages and in splenic macrophages. IFN-gamma also inhibited bacterial growth when added after infection of the macrophages, although the magnitude of the antibrucellae effects was less than that when it was added before infection. Furthermore, the maximal inhibitory effect was sustained only when IFN-gamma remained in the cultures after infection of the macrophages. PMID- 8418035 TI - Major stable peptides of Yersinia pestis synthesized during the low-calcium response. AB - It is established that the medically significant yersiniae require the presence of physiological levels of Ca2+ (ca. 2.5 mM) for sustained growth at 37 degrees C and that this nutritional requirement is mediated by a shared ca. 70-kb Lcr plasmid. The latter also encodes virulence factors (Yersinia outer membrane proteins [Yops] and V antigen) known to be selectively synthesized in vitro at 37 degrees C in Ca(2+)-deficient medium. In this study, cells of Yersinia pestis KIM were first starved for Ca2+ at 37 degrees C to prevent synthesis of bulk vegetative protein and then, after cell division had ceased, pulsed with [35S]methionine. After sufficient chase to ensure plasminogen activator-mediated degradation of Yops, the remaining major radioactive peptides were separated by conventional chromatographic methods and identified as Lcr plasmid-encoded V antigen and LcrH (and possibly LcrG), ca. 10-kb Pst plasmid-encoded pesticin and plasminogen activator, ca. 100-kb Tox plasmid-encoded fraction 1 (capsular) antigen and murine exotoxin, and chromosomally encoded antigen 4 (pH 6 antigen) and antigen 5 (a novel hemin-rich peptide possessing modest catalase activity but not superoxide dismutase activity). Also produced at high concentration was a chromosome-encoded GroEL-like chaperone protein. Accordingly, the transcriptional block preventing synthesis of bulk vegetative protein at 37 degrees C in Ca(2+) deficient medium may not apply to genes encoding virulence factors or to highly conserved GroEL (known in other species to utilize a secondary stress-induced sigma factor). PMID- 8418036 TI - T-lymphocyte response in a guinea pig model of tuberculous pleuritis. AB - The ability to induce tuberculous pleuritis in Mycobacterium bovis BCG-vaccinated guinea pigs was investigated as a model of human disease. A pleural effusion of 5 to 10 ml was obtained 6 to 7 days after the bilateral pleural injection of a suspension of heat-killed M. tuberculosis cells. Histological lesions were indicative of granulomatous pleuritis. Comparative studies of T lymphocytes obtained from pleural fluid and peripheral blood revealed increased antigen driven lymphoproliferation and E rosette formation in pleural effusion lymphocytes. The CD2+ T-lymphocyte population appeared to be expanded or concentrated in pleural fluid, suggesting a compartmentalization of antigen reactive T lymphocytes. These data demonstrate that experimental tuberculous pleuritis with effusion, closely resembling the human disease, can be produced in BCG-vaccinated guinea pigs. PMID- 8418037 TI - Candidate vaccine antigens identified by antibodies from mice vaccinated with 15- or 50-kilorad-irradiated cercariae of Schistosoma mansoni. AB - In murine schistosomiasis, the highest levels of resistance to cercarial challenge are obtained by vaccination with radiation-attenuated cercariae. To identify candidate vaccine antigens relevant to the vaccine model, we examined parasite antigens recognized by antibodies from mice vaccinated with irradiated cercariae of Schistosoma mansoni. To optimize recognition of a wide spectrum of antigens, several factors that influence the level of protection in this model were varied; specifically, we examined the effect of (i) single versus multiple vaccinations with irradiated cercariae, (ii) the dose of irradiation (15 or 50 kilorads) administered to the cercariae, and (iii) the genetic background of mouse strains, high-responder (C57BL/6J) versus moderate-responder (CBA/J) mice. We found that the number of vaccinations did not alter antibody specificity but modified the relative antibody titers against particular antigens. The dose of irradiation used to attenuate the immunizing cercariae had a similar effect on antibody titers but in addition influenced antibody specificity. Only mice that had been vaccinated with moderately irradiated cercariae recognized cathepsin B (Sm31) and Sm32. Interestingly, when vaccinated mice of the two strains, C57BL/6J and CBA/J, were compared, differences in antibody responses to particular antigens were observed. Both strains recognized the integral membrane protein Sm23, glutathione S-transferase, and cathepsin B, whereas Sm32 and paramyosin were recognized only by CBA/J mice, and heat shock protein 70 was recognized exclusively by C57BL/6J mice. In this study, we conclusively identified six distinct antigens that are specifically recognized by the humoral immune response of vaccinated mice. PMID- 8418038 TI - Listeria ivanovii is capable of cell-to-cell spread involving actin polymerization. AB - Listeria ivanovii has been considered to be pathogenic to animals but has rarely been found associated with human infections. It has been claimed that L. ivanovii lacks the actA gene, which in L. monocytogenes encodes a protein required for interaction with host cell actin. Using fluorescence microscopy and electron microscopy, we demonstrate that L. ivanovii can invade mammalian cells, lyse the phagosomal membrane, polymerize host cell actin, reorganize actin to form tails, and spread from cell to cell. However, no DNA homologous to the actA gene could be detected by polymerase chain reaction. Further, L. ivanovii lacks the 90-kDa surface protein which in L. monocytogenes is encoded by actA. Despite the ability to spread from cell to cell, L. ivanovii differed significantly from L. monocytogenes in being unable to form plaques on monolayers of 3T3 fibroblast cells. PMID- 8418039 TI - Structural and serological specificities of Pasteurella haemolytica lipopolysaccharides. AB - Lipopolysaccharides (LPSs) from 16 serotypes of Pasteurella haemolytica were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and examined by silver staining and immunoblotting. Silver staining of proteinase K digested cell lysates revealed two rough LPS serotypes (serotypes 2 and 8), which lacked demonstrable O-polysaccharide, while 14 others demonstrated a ladder pattern characteristic of smooth-type LPS. Purified LPSs from several serotypes yielded O-polysaccharide in addition to low-molecular-weight core oligosaccharide components when subjected to mild acid hydrolysis. Nuclear magnetic resonance spectroscopy revealed the O-chain polysaccharides of serotypes 1, 6, and 9 to be identical. Immunoblots using hyperimmune rabbit, mouse, bovine, and ovine sera from homologous and heterologous serotypes supported this finding and suggested that most of the A biotypes share common O-chain epitopes. Immunoblotting results also supported structural data which demonstrated that the O-polysaccharides of serotypes 3 and 15 and of serotypes 4 and 10 (T biotypes) are identical. Nuclear magnetic resonance analysis indicated that the core oligosaccharides of serotypes 1, 6, 8, 9, and 12 share similar structures, but that they are distinct from those of serotypes 3, 4, 10, and 15. Immunoblots with hyperimmune antisera and monoclonal antibody having specificity for the core region of serotype 1 LPS revealed shared epitopes in the core oligosaccharides of several A biotypes. Characterization of the molecular structure and antigenic specificities of LPS has been an important consideration in the development of purity and potency assays for veterinary vaccines which contain P. haemolytica. PMID- 8418040 TI - Comparison of aminopeptidase activities in four strains of mutans group oral streptococci. AB - In this study, native cells of Streptococcus mutans VA-29R and Streptococcus rattus FA-1 displayed significantly higher aminopeptidase activity than did cells of Streptococcus cricetus AHT or Streptococcus sobrinus 6715 toward the nitroanilide derivatives of leucine, alanine, methionine, arginine, and lysine. These differences in cellular aminopeptidase activity led us to investigate the subcellular localization of the aminopeptidase in these mutans group streptococci. Following conversion of native cells to protoplasts by treatment with lysozyme, most of the aminopeptidase activity detected in the native-cell preparations remained associated with the intact protoplasts. After lysis of protoplasts and differential centrifugation, most of the total cellular aminopeptidase activity was recovered with the cytoplasmic fraction. Membrane associated aminopeptidases represented only minor activities in these mutans group streptococci. Although the four strains showed no differences with respect to a predominant cytoplasmic localization for the aminopeptidase activities, the levels of activity in the cytoplasmic fractions from S. cricetus AHT and S. sobrinus 6715 were significantly lower than those measurable in the corresponding fractions from S. mutans VA-29R and S. rattus FA-1. These results support the conclusion that the differences in aminopeptidase activity expressed by these streptococci reflect quantitative differences rather than differences in enzyme subcellular localization. PMID- 8418042 TI - Patterns of cytokine production by mycobacterium-reactive human T-cell clones. AB - To gain insight into the functional capacity of human T cells in the immune response against Mycobacterium tuberculosis, we evaluated the spectrum of cytokines produced by mycobacterium-reactive human T-cell clones. Nine of 11 T cell clones bearing alpha beta or gamma delta T-cell receptors produced both Th1 and Th2 cytokines, a pattern resembling that of murine Th0 clones. The most frequent pattern was secretion of gamma interferon, tumor necrosis factor alpha (TNF), and interleukin-10 (IL-10), in combination with IL-2, IL-5, or both. Two clones produced only Th1 cytokines, and none produced exclusively Th2 cytokines. Although IL-4 was not detected in cell culture supernatants, IL-4 mRNA was detected by polymerase chain reaction amplification in two of six clones. There were no differences between the cytokine profiles of alpha beta and gamma delta T cells. A striking finding was the markedly elevated concentrations of TNF in clone supernatants, independent of the other cytokines produced. Supernatants from mycobacterium-stimulated T-cell clones, in combination with granulocyte macrophage colony-stimulating factor, induced aggregation of bone-marrow-derived macrophages, and this effect was abrogated by antibodies to TNF. The addition of recombinant TNF to granulocyte-macrophage colony-stimulating factor markedly enhanced macrophage aggregation, indicating that TNF produced by T cells may be an important costimulus for the granulomatous host response to mycobacteria. The cytokines produced by T cells may exert immunoregulatory and immunopathologic effects and thus mediate some of the clinical manifestations of tuberculosis. PMID- 8418041 TI - Meningococcal lipopolysaccharide (LPS)-derived oligosaccharide-protein conjugates evoke outer membrane protein- but not LPS-specific bactericidal antibodies in mice: influence of adjuvants. AB - Meningococcal lipopolysaccharide (LPS)-derived oligosaccharides (OS) were coupled to tetanus toxoid (TT) and purified P1.7,16 outer membrane proteins (OMP). The immunogenicities of the conjugates with and without the addition of the adjuvant Quil A or the nonionic block polymer L121 were studied in mice. Immunotype L2 and L3,7,9 OS-TT conjugates induced immunoglobulin G (IgG) responses that were strongly augmented by Quil A and L121. These adjuvants not only enhanced the amount of IgG evoked but also shifted the IgG subclass distribution from mainly IgG1 toward the complement-activating subclasses IgG2a and IgG2b. The antibodies induced were directed against the OS part of meningococcal LPS. They were not bactericidal for group B meningococci. Both the L3,7,9 OS-P1.7,16 OMP conjugate and purified P1.7,16 OMP evoked a strong IgG response against the P1.7,16 OMP but not against the L3,7,9 LPS. These anti-OMP IgG responses were comparable to the IgG OMP-specific responses induced by the H44/76 or HIII-5 outer membrane vesicles but still did not lyse group B meningococcal strains. The IgG response evoked with OS-OMP or purified OMP consisted mainly of the IgG1 subclass, whereas the H44/76 or HIII-5 outer membrane vesicles induced high amounts of bactericidal IgG2a and IgG2b antibodies next to the IgG1 antibodies. The addition of the adjuvant Quil A or L121 to OS-OMP or OMP resulted in the induction of high levels of bactericidal anti-P1.7,16-specific OMP antibodies, as reflected by the presence of substantial amounts of IgG2a and IgG2b antibodies. These results indicate that (i) mouse anti-LPS antibodies evoked by LPS-derived OS-protein conjugates are not bactericidal for group B meningococci, (ii) extensive purification of P1.7,16 OMP can lead to the loss of the intrinsic adjuvant properties of outer membrane vesicle preparations, and (iii) the addition of suitable adjuvants restores the ability of these purified P1.7,16 OMP to induce bactericidal antibodies. PMID- 8418043 TI - Nucleotide sequence of the variable domains within the major outer membrane protein gene from serovariants of Chlamydia trachomatis. AB - During studies in which we serotyped large numbers of Chlamydia trachomatis clinical isolates by using monoclonal antibodies, three novel serological variants (D-, D*, and I-) were identified. To determine the molecular basis for the altered monoclonal antibody reactions of these strains and other previously identified variants (Da, Ia, and L2a), we determined the nucleotide sequences of the variable domains in their major outer membrane protein genes. Da, D-, and D* variants differed by a single nucleotide and also an amino acid in the carboxy terminus of variable domain IV (VDIV) from the D serovar. The L2a variant also differed from L2 by a single amino acid but in VDII. Ia variants differed in VDI, III, and IV and I- variants differed in all four VDs from the I serovar. These studies demonstrate the potential for using major outer membrane protein VD sequencing as a highly sensitive typing method and further identify immunologically reactive major outer membrane protein epitopes. PMID- 8418044 TI - Cytokine appearance and effects of anti-tumor necrosis factor alpha antibodies in a neonatal rat model of group B streptococcal infection. AB - Cytokines are suspected of playing an important role in the pathophysiology of septic shock. This study was undertaken to determine whether tumor necrosis factor alpha (TNF-alpha) induces the production of other cytokines and mediates mortality in a neonatal rat model of sepsis caused by group B streptococci (GBS). We have measured TNF-alpha, interleukin-1 alpha (IL-1 alpha), interleukin-6 (IL 6), and gamma interferon (IFN-gamma) levels in neonatal rats infected with different strains (H738, 259, and 90) and doses (1 50% lethal dose [LD50] and 5 90% lethal doses [LD90]) of type III GBS. TNF-alpha and IL-6 were detected by the L929 cytotoxicity and the B9 proliferation assays, respectively, in serial plasma samples. IL-1 alpha and IFN-gamma were measured in spleen homogenates by enzyme linked immunosorbent assay kits by using antibodies raised against the corresponding mouse cytokines. Plasma TNF-alpha levels significantly rose above baseline values within 12 h after intraperitoneal challenge with 5 LD90 of GBS strain H738, corresponding to 3 x 10(3) CFU. A mean peak TNF-alpha concentration of 232 +/- 124 U/ml was reached at 20 h. Peak IL-1 alpha and IL-6 levels of 766 +/- 404 U/g and 1,033 +/- 520 U/ml, respectively, were reached at 24 h after bacterial challenge. Maximal spleen concentrations of IFN-gamma (449 +/- 283 U/g) were measured at 36 h. Concentrations of TNF-alpha, but not other cytokines, remained significantly elevated at 72 h, a time when mortality approached 100%. Significant correlations were found between concentrations of each of the cytokines tested and the logs of CFU concentrations in the blood. In order to ascertain whether TNF-alpha influenced the production of other cytokines, rat pups received two injections of anti-murine TNF-alpha or normal rabbit serum at 2 h before and at 26 h after challenge with live GBS. Plasma TNF-alpha bioactivity was undetectable in anti-TNF-alpha-treated animals, while IL-6 and IFN-gamma, but not IL-1 alpha, levels were significantly reduced, compared with normal serum controls. Rat pups pretreated with anti-TNF-alpha serum and infected with 1 and 5 LD90 of strains H738 and 259 showed enhanced early (48 to 72 h) survival. However, by 96 h this protection was no longer apparent. PMID- 8418045 TI - Association between virulence of Yersinia pestis and suppression of gamma interferon and tumor necrosis factor alpha. AB - It is established that Yersinia pestis, the causative agent of bubonic plague, and enteropathogenic Yersinia pseudotuberculosis and Yersinia enterocolitica share a ca. 70-kb low-calcium response or Lcr plasmid (Lcr+). The latter is known to encode regulatory functions that restrict growth at 37 degrees C in Ca(2+) deficient medium and virulence factors that are expressed only in vitro within this environment (e.g., certain Yops and V antigen). In this study, gamma interferon (IFN-gamma) was never detected in mice infected with 10 minimum lethal doses (MLD) of Lcr+ cells of Y. pestis, and significant levels of tumor necrosis factor alpha (TNF-alpha) arose only prior to death. Prompt and marked synthesis of these cytokines was observed upon infection with avirulent Lcr- mutants. Treatment of mice with exogenous IFN-gamma plus TNF-alpha inhibited multiplication of Lcr+ yersiniae in vivo, thereby providing protection against challenge with 10 MLD. Administration of both cytokines was required for absolute survival, suggesting a synergistic rather than cumulative interaction. This protective effect entailed cytokine priming as judged by subsequent detection of substantial levels of endogenous IFN-gamma and TNF-alpha. Monospecific anti-V antigen, known to provide passive immunity against 10 MLD of Lcr+ Y. pestis, permitted significant synthesis of endogenous IFN-gamma and TNF-alpha. These findings demonstrate that Lcr+ yersiniae suppress synthesis of cytokines and suggest that this effect is mediated by one or more Lcr plasmid-encoded virulence factors. PMID- 8418046 TI - Neuraminidase production by a Pasteurella haemolytica A1 strain associated with bovine pneumonia. AB - The properties of an extracellular neuroaminidase produced by a Pasteurella haemolytica A1 strain (isolated from a case of bovine pneumonia) during growth in a defined medium were examined in this investigation. This enzyme, isolated from concentrated culture supernatants of P. haemolytica A1, was active against N acetylneuramin lactose, human alpha 1-acid glycoprotein, fetuin, and bovine submaxillary mucin. Neuraminidase production paralleled bacterial growth in a defined medium and was maximal in the stationary phase of growth. The enzyme was purified to homogeneity by a combination of salt fractionation, ion-exchange chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200. These procedures yielded an enzyme preparation that possessed a specific activity of 100.62 mumol of sialic acid released per min per mg of protein against human alpha 1-acid glycoprotein. The Km value for this enzyme with human alpha 1-acid glycoprotein as the substrate was 1.1 mg/ml, and the enzyme possessed a pH optimum of 6.5. The P. haemolytica A1 neuraminidase had a molecular weight of approximately 150,000 as estimated by gel filtration and approximately 170,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was stable at 4 degrees C for 3 h. At 37 degrees C for 3 h, 25% of enzymatic activity was lost. Approximately 55% of the enzyme activity was lost within 30 min at 50 degrees C, with greater than 70% of the enzyme activity being destroyed within 10 min at temperatures of > or = 65 degrees C. PMID- 8418047 TI - Expression of the Mycobacterium tuberculosis 19-kilodalton antigen in Mycobacterium smegmatis: immunological analysis and evidence of glycosylation. AB - The gene encoding a 19-kDa antigen from Mycobacterium tuberculosis was expressed as a recombinant protein in the rapid-growing species Mycobacterium smegmatis. The recombinant antigen was expressed at a level approximately ninefold higher than in M. tuberculosis and, like the native antigen, was found in the pellet fraction after high-speed centrifugation of bacterial extracts. The 19-kDa antigen in crude bacterial extracts, and the purified recombinant antigen, bound strongly to concanavalin A, indicating the possibility of posttranslational glycosylation. The recombinant antigen stimulated T-cell proliferation in vitro when added to assays either in the form of whole recombinant bacteria or as a purified protein. Homologous expression of mycobacterial antigens in a rapid growing mycobacterial host may be particularly useful for the immunological characterization of proteins which are subject to posttranslational modification. PMID- 8418048 TI - Specific T-cell recognition of the merozoite proteins rhoptry-associated protein 1 and erythrocyte-binding antigen 1 of Plasmodium falciparum. AB - The merozoite proteins merozoite surface protein 1 (MSP-1) and rhoptry-associated protein 1 (RAP-1) and synthetic peptides containing sequences of MSP-1, RAP-1, and erythrocyte-binding antigen 1, induced in vitro proliferative responses of lymphocytes collected from Ghanaian blood donors living in an area with a high rate of transmission of malaria. Lymphocytes from a large proportion of the Ghanaian blood donors proliferated in response to the RAP-1 peptide, unlike those of Danish control blood donors, indicating that this sequence contains a malaria specific T-cell epitope broadly recognized by individuals living in an area with a high transmission rate of malaria. Most of the donor plasma samples tested contained immunoglobulin G (IgG) and IgM antibodies recognizing the merozoite proteins, while only a minority showed high IgG reactivity to the synthetic peptides. PMID- 8418049 TI - Ultrastructural analysis of growth of Nocardia asteroides during invasion of the murine brain. AB - BALB/c mice were infected with 10(6) CFU of log-phase cells of Nocardia asteroides GUH-2 by tail vein injection (at this lethal inoculum dose, approximately 800 to 1,000 CFU becomes deposited in the brain). At 24 h after infection, the ultrastructural interactions of the nocardiae during growth within the murine brain were investigated. The nocardiae grew perivascularly in the pons, substantia nigra, hypothalamus, and thalamus portions of the brain, where they were either within or associated with most brain cell types. There appeared to be a propensity for growth within the soma of neurons and their axonal extensions. The nocardial cells were surrounded by 1 to 30 layers of membrane, and the innermost membrane was usually tightly adherent to the cell wall. This compartmentalization of nocardiae within brain cells could contribute to their failure to induce an inflammatory response or a cytopathic effect. Nevertheless, the bacteria were able to obtain adequate nutrients from the host to grow within the brain. The nocardiae were not completely inert, since some of the brain cells showed signs of degeneration. The myelin sheaths of axons were the most strongly affected, and there was evidence of demyelinization and axonal degeneration. Nocardiae growing within brain cells were phagocytized by compact, dense cells that were probably microglia. There was no ultrastructural evidence that the nocardiae were damaged by these phagocytes 24 h after infection; nevertheless, these cells may be important for the elimination of nocardiae from the brain during a nonlethal infection. PMID- 8418050 TI - Effects of N-acetylglucosamine on carbohydrate fermentation by Streptococcus mutans NCTC 10449 and Streptococcus sobrinus SL-1. AB - We have investigated the ability of two species of streptococci isolated from the human oral cavity (Streptococcus mutans NCTC 10449 and Streptococcus sobrinus SL 1) to metabolize N-acetylglucosamine (GlcNAc), a naturally occurring amino sugar present in saliva and human glycoproteins, when provided as the sole fermentable carbohydrate and determined the effects of the presence of GlcNAc on the fermentation of other carbohydrates. S. mutans used GlcNAc at concentrations of up to 10 mM to increase cell numbers, but S. sobrinus was unable to ferment the amino sugar alone and its uptake only occurred in the presence of a fermentable carbohydrate. GlcNAc had a marked inhibitory effect on the ability of S. sobrinus to produce lactic acid from glucose, sucrose, and fructose, at the same time increasing the lag period and doubling time of batch-grown cells. Such patterns of inhibition were found with S. mutans, but the effects were less than those seen in S. sobrinus. In mixed culture studies of the two species, S. sobrinus became the predominant organism when 10 mM glucose was supplied as the sole fermentable carbohydrate, with a concomitant decrease in the numbers of S. mutans cells, but supplementation of the broth with 10 mM glucose and 10 mM GlcNAc resulted in the emergence of S. mutans as the predominant organism. S. mutans and S. sobrinus grown in media containing glucose possessed the ability to transport glucose and GlcNAc, probably via the same glucose-phosphotransferase system at similar rates. However, intracellular levels of N-acetylglucosamine-6-phosphate deacetylase and glucosamine-6-phosphate deaminase were markedly higher in S. mutans grown on glucose and GlcNAc than in S. sobrinus: 34 and 398 and 8 and 17 nmol of NADPH formed per mi per mg of protein for S. mutans and S. sobrinus, respectively. We propose that GlcNAc inhibited growth of S. sobrinus in media containing glucose and GlcNAc by competing with glucose for the glucose phosphotransferase, depleting intracellular levels of phosphoenolpyruvate, and possessing, in contrast to S. mutans, low levels of N-acetyl-glucosamine-6 phosphate deacetylase and glucosamine-6-phosphate deaminase activity. Together, these data suggest that in dental plaque, S. sobrinus when exposed to GlcNAc will have a reduced ability to compete with S. mutans for dietary carbohydrates, contributing to the greater frequency of isolation of S. mutans from human populations. PMID- 8418051 TI - Apathogenic, intestinal, segmented, filamentous bacteria stimulate the mucosal immune system of mice. AB - Segmented filamentous bacteria (SFBs) are apathogenic autochthonous bacteria in the murine small intestine that preferentially attach to Peyer's patch epithelium. SFBs have never been cultured in vitro. We have studied the effects of SFBs on the immune system of the host. Mice monoassociated with SFBs were compared with germ-free mice and with mice without SFBs but with a specific pathogen-free (SPF) gut flora. SFBs versus no microbial flora raised the number of lymphoid cells in the lamina propria of the ileal and cecal mucosa, raised the number of immunoglobulin A (IgA)-secreting cells in the intestinal mucosa, produced elevated IgA titers in serum and intestinal secretions, and enhanced the concanavalin A-induced proliferative responses of mesenteric lymph node cells. The SPF flora had effects similar to but less pronounced than those mediated by SFBs. The results indicate that SFBs stimulate the mucosal immune system to a greater extent than do other autochthonous gut bacteria. PMID- 8418052 TI - Purification and characterization of exoenzyme S from Pseudomonas aeruginosa 388. AB - Exoenzyme S was purified > 1,500-fold from the culture supernatant fluid of Pseudomonas aeruginosa 388 at high yield without utilization of solvents or detergents. Two proteins, with apparent molecular sizes of 53 and 49 kDa, cofractionated with exoenzyme S activity. Rabbit anti-49-kDa-protein immunoglobulin G was prepared by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis-purified 49-kDa protein as immunogen. Anti-49-kDa-protein IgG inhibited the ADP-ribosyltransferase activity of purified exoenzyme S in a dose dependent manner, which indicated a role for the 49-kDa protein in the ADP ribosylation reaction. Analysis by ultrafiltration showed that exoenzyme S activity and the 53- and 49-kDa proteins cofractionated and that exoenzyme S was apparently > 300 kDa in size. Urea (8 M) and 1.0% Triton X-100 reversibly decreased the apparent molecular sizes of exoenzyme S activity and the 53- and 49 kDa proteins to between 30 and 100 kDa. PMID- 8418053 TI - Induction of mucosal immunity by intranasal application of a streptococcal surface protein antigen with the cholera toxin B subunit. AB - The level and distribution of isotype-specific antibodies in various secretions and of antibody-secreting cells in corresponding lymphoid organs and tissues were compared in mice immunized with Streptococcus mutans surface protein antigen I/II (AgI/II) conjugated to the cholera toxin B subunit (CTB), given intranasally (i.n.) or intragastrically (i.g.), with or without free cholera toxin (CT) as an adjuvant. Immunization i.n. induced stronger initial antibody responses to AgI/II in both serum and saliva than immunization i.g., but salivary immunoglobulin A (IgA)-specific antibody responses to immunization about 3 months later were not increased relative to total salivary IgA concentrations. Specific antibodies induced by i.n. immunization were as widely distributed in serum, saliva, tracheal wash, gut wash, and vaginal wash as those induced by i.g. immunization. Likewise, specific antibody-secreting cells were generated in the spleen, salivary glands, intestinal lamina propria, and mesenteric and cervical lymph nodes by either route of immunization. The strongest salivary IgA antibody response was induced by AgI/II-CTB conjugate given i.n., but the addition of CT did not further enhance it. However, free CTB could effectively replace CT as an adjuvant in i.n. immunization with unconjugated AgI/II. Booster i.n. immunization with AgI/II plus either free CT or CTB induced stronger recall serum antibody responses than conjugated AgI/II-CTB with or without CT as an adjuvant. Therefore, i.n. immunization with a protein antigen and free or coupled CTB is an effective means of generating IgA antibody responses expressed at several mucosal sites where protective immunity may be beneficial. PMID- 8418054 TI - Storage reservoirs of hemin and inorganic iron in Yersinia pestis. AB - It is established that a high-frequency chromosomal deletion of ca. 100 kb accounts for the loss of properties making up the pigmented phenotype (Pgm+) of wild-type Yersinia pestis. These determinants are known to include virulence by peripheral routes of injection, sensitivity to the bacteriocin pesticin, adsorption of exogenous hemin or Congo red at 26 degrees C, and growth in iron sequestered medium at 37 degrees C. We have now identified the outer membrane as the primary site of exogenous hemin storage in Pgm+ cells grown at 26 degrees C. Significant outer membrane storage of hemin did not occur in Pgm- mutants or in Pgm+ cells cultivated at 37 degrees C. However, both Pgm+ and Pgm- organisms grown at 37 degrees C contained a periplasmic reservoir of hemin, which may be associated with a temperature-dependent ca. 70-kDa peptide recently equated with antigen 5. At 37 degrees C, Pgm+ and Pgm- yersiniae also utilized a cytoplasmic ca. 19-kDa bacterioferritin-like peptide for deposition of inorganic iron. Incorporation of [55Fe]hemin into pools at 37 degrees C was not significantly inhibited by competition with excess unlabeled Fe3+. However, excess unlabeled hemin modestly competed with incorporation of label from 55FeCl3. This relative independence of storage pools observed at 37 degrees C is consistent with physiological linkage to in vivo acquisition and transport of Fe3+ from ferritin and of hemin from hemoglobin, myoglobin, or hemopexin. PMID- 8418055 TI - Production, characterization, and application of monoclonal antibodies which distinguish three glucosyltransferases from Streptococcus mutans. AB - Thirty-three murine monoclonal antibodies (MAbs) against the three glucosyltransferases (GTFs) (GTF-I, -SI, and -S) from Streptococcus mutans were obtained by the fusion of murine myeloma cells (P3X63-Ag8-U1) with spleen cells of BALB/c mice immunized with pure GTF-S or partially purified GTF-I from serotype c S. mutans PS14. The immunoreactivities of these MAbs were tested by enzyme-linked immunosorbent assay and Western blotting (immunoblotting) with various GTF preparations. GTF-I and GTF-SI were expressed from two Streptococcus milleri or Escherichia coli transformants harboring gtfB or gtfC, respectively. All of the five MAbs raised against the GTF-S from PS14 reacted only with the homologous enzymes. Of these, 8 MAbs reacted only with the gtfB gene product (GTF I), 4 MAbs reacted only with the gtfC gene product (GTF-SI), and the remaining 16 MAbs reacted with both gene products. The existence of GTF-SI in the purified GTF I from PS14 was demonstrated by Western blot analysis using the representative monospecific MAbs. Further, the relative levels of the three GTFs in the extracellular and cellular fractions of S. mutans clinical isolates were examined by immunoblot analysis. The findings indicated that the relative level of GTF-SI, unlike that of GTF-I or GTF-S, differed markedly among isolates although the three GTFs were synthesized extracellularly by all the strains. PMID- 8418056 TI - Relative avidity of serum immunoglobulin G antibody for the fimbria antigen of Actinobacillus actinomycetemcomitans in patients with adult periodontitis. AB - The relative avidity of serum immunoglobulin G antibody for the fimbria antigen of Actinobacillus actinomycetemcomitans was assessed by diethylamine dissociation enzyme-linked immunosorbent assay in patients with adult periodontitis. High titer sera from patients not harboring A. actinomycetemcomitans had significantly higher avidities for the fimbria antigen than did high-titer sera from patients with A. actinomycetemcomitans in their periodontal pockets. The elicited antibodies against the fimbria antigen may afford protection against A. actinomycetemcomitans infection. PMID- 8418057 TI - Binding of pertussis toxin to lipid vesicles containing glycolipids. AB - The binding of pertussis toxin and its B oligomer to lipid vesicles containing glycosphingolipids was studied. Both pertussis toxin and the B oligomer bound to lipid vesicles containing ganglioside GD1a. Binding of pertussis toxin to these vesicles decreased upon treatment of the vesicles with neuraminidase, suggesting that sialic acid residues are important for efficient binding of the toxin to GD1a. PMID- 8418058 TI - Inhibition of growth of Mycobacterium avium in murine and human mononuclear phagocytes by migration inhibitory factor. AB - Infections caused by Mycobacterium avium, the most common form of diseminated bacterial disease in AIDS patients, are difficult to treat because of their resistance to many antimycobacterial drugs. The results of the present study show that recombinant migration inhibitory factor, a 12-kDa molecule recently isolated by COS-1 cell expression screening of cDNA from a human T-cell hybridoma, has potent inhibitory activity on the growth of a panel of clinical isolates of M. avium within both bone-marrow-derived murine macrophages and cultured human blood monocytes. These cells cultured in recombinant migration inhibitory factor exhibit various signs of activation, including cell division, morphological changes such as evidence of substantial phagolysosomal fusion, and enhanced secretion of tumor necrosis factor. PMID- 8418059 TI - Interactions of Nocardia asteroides with murine glia cells in culture. AB - Astrocytes and microglia were obtained from brains of newborn mice and infected with Nocardia asteroides. Scanning electron micrographs showed nocardiae adhering to the astrocyte cell surface and entering the cytoplasm. After 6 h of incubation the intracellular nocardiae appeared as filaments, demonstrating that growth was occurring. In contrast, the microglia phagocytized nocardiae, but after 6 h the presence of coccoid cells indicated that nocardial growth was inhibited. PMID- 8418060 TI - Transgenic mice expressing C-reactive protein are susceptible to infection with Plasmodium yoelii sporozoites. AB - Human and rat C-reactive proteins, major acute-phase reactants, bind to sporozoites and inhibit their in vitro development in hepatocytes (A. Nussler, S. Pied, M. Pontet, F. Miltgen, L. Renia, M. Gentilini, and D. Mazier, Exp. Parasitol. 72:1-7, 1991, and S. Pied, A. Nussler, M. Pontet, F. Miltgen, H. Matile, P.-H. Lambert, and D. Mazier, Infect. Immun. 57:278-282, 1989). We show here that rabbit C-reactive protein has identical properties. Nevertheless, infection by Plasmodium yoelii sporozoites was not prevented in transgenic mice engineered to express rabbit C-reactive protein following induction of gluconeogenesis. PMID- 8418061 TI - Adherence of Helicobacter pylori to cultured human gastric epithelial cells. AB - Experiments were performed to demonstrate that adherence of Helicobacter pylori to gastric epithelial cells causes alterations in the cell cytoskeleton. H. pylori intimately attached to cultured human gastric epithelial cells on small cellular projections, while there was no intimate association of H. pylori with cultured human esophageal epithelial cells. Fluorescein-conjugated phalloidin staining of gastric epithelial cells showed that H. pylori adherence stimulated actin polymerization; this stimulation was not observed with esophageal cells. Also, this organism's selectivity for gastric mucosa was supported by rare binding of bacteria to esophageal epithelial cells and gastric fibroblasts. PMID- 8418062 TI - Rat model of congenital toxoplasmosis: rate of transmission of three Toxoplasma gondii strains to fetuses and protective effect of a chronic infection. AB - The incidence of congenital toxoplasmosis in Fischer rats infected between the 8th and 12th days of pregnancy with three different strains of Toxoplasma gondii (RH, 76K, and Prugniaud) were 58.2, 35.2, and 62.8%, respectively. No infected fetuses were collected from rats previously infected with RH or Prugniaud strain parasites, even if the rats were reinfected during pregnancy. Since pups from chronically infected mothers are protected from congenital toxoplasmosis, rat infection could thus constitute a relevant model for immunological studies and vaccine design. PMID- 8418063 TI - Acid resistance in enteric bacteria. AB - Shigella species require a uniquely small inoculum for causing dysentery. One explanation for the low infective dose is that Shigella species are better able to survive the acidic conditions encountered in the stomach than are other enteric pathogens. We have tested Shigella species, Escherichia coli, and Salmonella species for the ability to survive at pH 2.5 for at least 2 h. Most isolates of Shigella and E. coli survived this treatment, whereas none of the Salmonella isolates were able to do so. The ability of Shigella species to survive at low pHs does not require the presence of the large virulence plasmid or growth at 37 degrees C but is strikingly dependent on growth phase. We have also found that Shigella isolates exposed to acid lose the ability to invade epithelial cells. PMID- 8418064 TI - A prospective study of the association between the human humoral immune response to Plasmodium falciparum blood stage antigen gp190 and control of malarial infections. AB - The human humoral immune response to the Plasmodium falciparum merozoite surface antigen gp190 was analyzed to determine the rate of reinfection by the parasite and the ability to control parasite density. The prospective study was carried out in a West African village where malaria is hyperendemic. No correlation between the antibody titers and protection against infection was observed within the group of children. Positive and negative associations of antibody specificities with protection against and/or control of parasitemia were, however, found for adolescents and adults, respectively. Thus, in adolescents, the presence of antibodies to gp190 fragment M6 correlates with a 50% reduced risk of P. falciparum infection and an increased ability to control parasitemia, whereas in adults, the humoral response to some of the polymorphic regions of gp190 associates with an increased risk of infection. PMID- 8418065 TI - Comparative effectiveness of the cholera toxin B subunit and alkaline phosphatase as carriers for oral vaccines. AB - The purpose of this study was to determine whether the B subunit of cholera toxin (CtxB) has adjuvant activity over and above serving as a carrier protein for orally administered vaccines. An oligonucleotide that encodes an antigenic determinant (GtfB.1) from the glucosyltransferase B gene (gtfB) of Streptococcus mutans was genetically fused to the 5' terminus of either the CtxB gene (ctxB) or the Escherichia coli alkaline phosphatase gene (phoA). The resulting chimeric proteins were expressed in a phoA mutant strain of E. coli and then purified. The antigenicities of the proteins were confirmed by immunoblotting analysis using antisera specific for GtfB, CtxB, or PhoA. An equimolar amount of peptide on each carrier was administered by gastric intubation to mice three times at 10-day intervals. Antibody titers to the peptide, CtxB, and PhoA (in the serum, intestine, vagina, saliva, and bronchus) were determined by enzyme immunoassay. Antibody to the peptide was detected only in the sera of mice immunized with the peptide fused to CtxB. No antipeptide antibody was detected in mice immunized with the peptide fused to PhoA. The lack of detectable levels of antipeptide antibody in intestinal lavage fluid was attributed to dilution of the sample beyond the sensitivity of the assay. This was confirmed by cultivation of Peyer's patch and mesenteric lymph node tissue from mice orally immunized with the GtfB.1::CtxB chimera. Using this method, antipeptide antibody was detected in the culture fluid. We conclude that CtxB possesses unique properties that allow it to act as more than a simple carrier protein. PMID- 8418066 TI - YopB and YopD constitute a novel class of Yersinia Yop proteins. AB - Virulent Yersinia species harbor a common plasmid that encodes essential virulence determinants (Yersinia outer proteins [Yops]), which are regulated by the extracellular stimuli Ca2+ and temperature. The V-antigen-encoding operon has been shown to be involved in the Ca(2+)-regulated negative pathway. The genetic organization of the V-antigen operon and the sequence of the lcrGVH genes were recently presented. The V-antigen operon was shown to be a polycistronic operon having the gene order lcrGVH-yopBD (T. Bergman, S. Hakansson, A. Forsberg, L. Norlander, A. Macellaro, A. Backman, I. Bolin, and H. Wolf-Watz, J. Bacteriol. 173:1607-1616, 1991; S. B. Price, K. Y. Leung, S. S. Barve, and S. C. Straley, J. Bacteriol. 171:5646-5653, 1989). We present here the sequence of the distal part of the V-antigen operons of Yersinia pseudotuberculosis and Yersinia enterocolitica. The sequence information encompasses the yopB and yopD genes and a downstream region in both species. We conclude that the V-antigen operon ends with the yopD gene. This conclusion is strengthened by the observation of an insertion-like element downstream of the yopD gene. The translational start codons of YopB and YopD have been identified by N-terminal amino acid sequencing. By computer analysis, the yopB and yopD gene products were found to be possible transmembrane proteins, and YopD was shown to contain an amphipathic alpha-helix in its carboxy terminus. These findings contrast with the general globular pattern observed for other Yops. Homology between Yersinia LcrH and Shigella flexneri IppI and between Yersinia YopB and S. flexneri IpaB was found, suggesting conservation of this locus between these two genera. YopB was also found to have a moderate level of homology, especially within the hydrophobic regions, to members of the RTX protein family of alpha-hemolysins and leukotoxins, indicating that YopB might exhibit a similar function. PMID- 8418067 TI - Effect of pH and osmolality on in vitro phagocytosis and killing by neutrophils in urine. AB - Phagocytosis and intracellular killing of two strains of Escherichia coli and a Staphylococcus saprophyticus by polymorphonuclear neutrophils (PMN) in pooled sterile urine at three osmolalities (800, 485, and 200 mosM/kg of H2O) between pHs 5 and 8 was investigated. Urine at 800 mosM virtually abolished phagocytosis of both E. coli strains, regardless of pH, and reduced the phagocytosis of S. saprophyticus to 30%; no killing of any organisms took place at this osmolality. On the other hand, phagocytosis was a good in urine as in Hanks balanced salt solution at both 485 and 200 mosM between pHs 6 and 8. Phagocytosis of all three strains was virtually abolished at pH 5. Killing of the strains by PMN was optimal between pHs 6.5 and 7.5 in urine at 485 mosM (being at least 90% of the control values in Hanks balanced salt solution), whereas at 200 mosM killing was reduced to 50 to 70% of these values. Reduced killing of all three strains occurred at pH 8, whereas at pH 6 only S. saprophyticus was killed. Thus, the bactericidal activity of PMN in urine was more sensitive than phagocytic function to alterations in pH. The dominant modulating factor affecting PMN function in urine of 500 mosM or less was pH, but osmolality had a greater influence at 800 mosM. Thus, raising the pH of urine and reducing the osmolality may increase the ability of natural defense mechanisms to eliminate infecting organisms. PMID- 8418068 TI - Role of attached lipid in immunogenicity of Borrelia burgdorferi OspA. AB - OspA is a protective antigen of the Lyme disease spirochete Borrelia burgdorferi. Expression of the full-length B. burgdorferi B31 OspA gene in Escherichia coli produces a protein that is processed posttranslationally by signal peptidase II and contains an attached lipid moiety. The recombinant OspA lipoprotein has been purified by detergent extraction and ion-exchange chromatography. Priming and boosting with OspA lipoprotein, either with no adjuvant or adsorbed to alum, elicited a strong, dose-dependent immunoglobulin G response. Serum from vaccinated mice inhibited spirochetal growth in vitro. Mice immunized twice with as little as 0.4 micrograms of OspA lipoprotein were protected against an intradermal challenge with 10(4) infectious spirochetes. The ability of the purified recombinant lipoprotein to induce a strong protective response in the absence of toxic adjuvants makes it an excellent candidate antigen for a human vaccine against Lyme disease. By contrast, no serum immunoglobulin G or growth inhibitory response to OspA nonlipoprotein was seen at any dose. The difference in immunogenicities of the lipoprotein and nonlipoprotein forms of OspA is not due to any difference in the antigenicities of the two proteins. These results suggest that posttranslational lipid attachment is a critical determinant of the immunogenicity of the OspA protein. PMID- 8418069 TI - Purification and partial characterization of the major outer membrane protein of Haemophilus somnus. AB - We purified the major outer membrane protein (MOMP), which is the most abundant OMP (with an apparent molecular mass of 40 kDa), from Haemophilus somnus strain 8025. The method involves solubilization of the MOMP with Zwittergent 3-14 and further purification accomplished by ion-exchange and molecular-sieve chromatographies. The amino-terminal sequence of the MOMP showed considerable similarity to those of porin proteins from other gram-negative bacteria. The MOMP of H. somnus is immunogenic to rabbits and calves. Hyperimmune sera from rabbits and calves reacted with both the MOMP and lipopolysaccharides in enzyme-linked immunosorbent assay (ELISA) and immunoblot analysis. The rabbit antiserum to the MOMP was cross-reactive with whole-cell preparations from strains 8025, D1238, NT2301, and 540 at a band with a molecular mass of 40 kDa in immunoblot analysis, although the reactivity of the rabbit antiserum with strain 540 was lower than those with the other strains tested. Two murine monoclonal antibodies (MAbs) to the MOMP were developed. ELISA with the OMP fractions as the antigens showed that one MAb was cross-reactive with the four strains but that the other MAb was reactive with the three strains other than strain 540. These results indicate that the MOMP of H. somnus possesses at least two antigenic determinants and that the MOMP of strain 540 is antigenically different from those of the other strains. The antigenic heterogeneity of the H. somnus MOMP has implications regarding the development of a serotyping system with MAbs that is based on the MOMP epitopes. PMID- 8418071 TI - The political process and its relationship to the psychotherapy of substance misusers: an historical perspective. AB - There are massive changes underway in the allocation of funds for health care in the United States which will impact upon services provided for users and misusers of intoxicating substances. Recent findings suggest that conditions in the marketplace and the development of professions have effected standards of care rather than a reasoned analysis of need and outcome. Psychologists question to what extent they will be involved in public policy issues including what is clinically and socially relevant. The goal of this paper was to determine if an historical perspective upon federal regulation of intoxicating substances (tobacco/alcohol/drugs) would enlighten the psychotherapy scientist in the pursuit of standards for service. The methodology included a review of the economic and social structure of colonial America which included tobacco plantations, breweries, and distilleries as well as a review of the formation of the federal government and political system in which men, women, and slaves were each assigned different performance standards/roles and economic rewards within the community. The implication is that potential for self-regulation and psychological development is based upon the legacy of our forefathers. PMID- 8418072 TI - The relationship between emotional state and alcohol consumption. AB - Based on a November 1986 national probability sample of 1,069 American drinkers (i.e., people who drank at least one alcoholic beverage in the past 7 days) aged 21 and over, this study investigates the relationship between 10 emotional states (i.e., happy, sad, calm, tense, stimulated, bored, irritable, romantic, festive, and lonely) and the amount of beer, wine, distilled spirits, and/or wine cooler consumed during respondents' last drinking episode. The results indicate that: (1) Although all of the mediating variables studied affected the relationship between affect and alcohol consumption, age appears to be the most influential of these variables; and (2) For each alcoholic beverage type, at least one emotional state was significantly related to amount consumed. PMID- 8418070 TI - Interleukin-6 production in a murine model of Pneumocystis carinii pneumonia: relation to resistance and inflammatory response. AB - The production of interleukin-6 (IL-6) and its possible relationship to host resistance and inflammatory response to Pneumocystis carinii infection were examined in mice with severe combined immunodeficiency (SCID mice). IL-6 activity was detected in the serum and lungs of P. carinii-infected mice but not in mice free of P. carinii. Moreover, the IL-6 levels in P. carinii-infected mice increased markedly after spleen cell reconstitution but then decreased to an undetectable level after the clearance of P. carinii. However, neutralization of IL-6 activity in spleen cell-reconstituted SCID mice by treatment with anti-IL-6 immunoglobulin G (IgG) resulted in no significant effect on the clearance of P. carinii (P > 0.05). Both the serum and lungs of treated mice contained an excess amount of anti-IL-6 IgG and lacked detectable IL-6. These results suggest that failure to inhibit the P. carinii clearance by anti-IL-6 treatment was not due to insufficient administration of antibody or incomplete neutralization of IL-6 activity. However, compared with mice receiving rat control IgG, mice treated with anti-IL-6 IgG had significantly higher numbers of neutrophils and lymphocytes (particularly CD8+ cells) in the lung lavage fluids (P < 0.05 for both) at day 19 after reconstitution. In addition, the levels of both total IgG (P < 0.001) and P. carinii-specific antibodies (P < 0.05) in the serum of mice treated with anti-IL-6 were significantly higher than those in control mice. These results indicate that although P. carinii infection causes both local and systemic production of IL-6 in SCID mice, IL-6 does not appear to play a crucial role in the clearance of P. carinii. However, it appears that during resolution of P. carinii pneumonia, IL-6 plays a role in the regulation of pulmonary inflammation and antibody responses. PMID- 8418073 TI - Methadone maintenance clients in Amsterdam after five years. AB - The aim of the low-threshold Amsterdam Methadone Maintenance (MM) system is regulation of clients, i.e., the development of a way of life which is more acceptable to society and more satisfactory for clients. In this paper we describe the regulation of two random samples of clients who entered the Amsterdam MM 5 years earlier. Seventeen clients who left MM within 5 years are compared with 21 clients who stayed. The study indicates that clients who leave use less hard drugs and methadone, take better care of themselves, and have more often established a drug-free social network. PMID- 8418074 TI - Well-differentiated villoglandular adenocarcinoma of the uterine cervix: a clinicopathological study of 24 cases. AB - We reviewed the clinical and pathological features of 24 cases of well differentiated villoglandular adenocarcinoma of the uterine cervix. The patients' ages ranged from 27 to 54 years, with an average age of 37. At least 15 patients were taking oral contraceptive prior to diagnosis, compared to five of 18 in a control group of patients with various other histologic types of cervical adenocarcinoma. All of the neoplasms were exophytic polypoid lesions with thick or thin papillae lined by endocervical, endometrial or intestinal-type epithelium showing mild cytologic atypia. Ten were associated with adenocarcinoma in situ, eight with cervical intraepithelial neoplasia, and one with microglandular hyperplasia. All tumors were confined to the cervix. Five patients were treated by excisional biopsy or cone biopsy, four by simple hysterectomy with prior or subsequent radiation therapy, and 15 by radical hysterectomy. All patients are alive and well, with no evidence of recurrent disease. The follow-up ranged from 7 to 77 months, with a mean of 36 months. PMID- 8418075 TI - Subinvolution of the uteroplacental arteries: an immunohistochemical study. AB - Subinvolution of uteroplacental arteries is a well-recognized cause of hemorrhage in the postpartum period. Although the physiological changes in these arteries during pregnancy are well documented, the sequence of events in normal involution is largely unknown. A recent immunohistochemical study has raised the possibility of an abnormal interaction between maternal uterine cells and fetal trophoblast in subinvolution. An indirect immunoperoxidase technique was used to compare deposition of complement components and immunoglobulin in subinvoluted and normally involuted uteroplacental arteries in 25 cases of postpartum hemorrhage. Deposits of C1q, C3d, C4, and C9 were detected within the walls of normally involuted vessels, whereas deposition of C1q, C3d, and C4 was absent in subinvoluted vessels; C9 was detected only focally. Deposition of immunoglobulins G, A, and M mirrored those described for complement components. The results suggest that immunological factors are necessary for the process of normal involution of uteroplacental arteries and are deficient in subinvoluted vessels. PMID- 8418076 TI - Accuracy and survival benefit of cytological prediction of endometrial carcinoma on routine cervical smears. AB - Among 359 women who received a cytology report of endometrial malignancy from the Victorian Cytology Service during 1982-87, the positive predictive value for a later histological diagnosis of endometrial malignancy was 64%. The positive predictive value was significantly higher for the group of women in whom the cytopathologists made definite predictions of endometrial malignancy as compared to the group where the cytologic features were only suggestive of endometrial malignancy (75% vs. 50%, chi 2 = 23.4; p < 0.001). The sensitivity of cervical cytology performed within two years of the diagnosis of endometrial malignancy was 28%. The odds ratio of death from endometrial cancer among women where the cytology may have allowed for an early diagnosis in comparison to women where cytology did not hasten the diagnosis was 0.78 (95% confidence interval = 0.25 2.47; p > 0.05). We conclude that while cervical cytology can predict the presence of malignancy for a small proportion of women with endometrial cancer, there remains no evidence that a cervical cancer screening program will make a major impact on reducing the morbidity and mortality from endometrial cancer. PMID- 8418077 TI - Tumor-infiltrating lymphocytes from ovarian tumors of low malignant potential. AB - Tumor-infiltrating lymphocytes (TIL) from human ovarian tumors of low malignant potential (LMP) were studied in situ by immunohistology and characterized functionally after isolation from tumors. In comparison to ovarian carcinomas, borderline (LMP) ovarian tumors contained significantly fewer infiltrating leukocytes. Phenotypically, TIL from LMP tumors contained significantly fewer activated (HLA-DR+ or CD25+) lymphocytes than did fresh TIL from ovarian carcinomas. Also, percentages of CD3-CD56+ natural killer cells were higher in LMP than in ovarian carcinomas. TIL obtained from LMP tumors proliferated well in vitro in the presence of interleukin 2 (IL2) and tumor necrosis factor alpha (TNF alpha) but did not show a selective enrichment in CD3+CD8+T lymphocytes generally observed with TIL from ovarian carcinomas. Antitumor cytotoxicity against a panel of tumor cell targets of cultured TIL from LMP tumor did not parallel that of effectors generated from autologous peripheral blood lymphocytes. However, no significant enrichment in reactivity against autologous tumor cells was observed in long-term cultures of TIL from LMP tumors. Thus, considerable differences were observed between phenotypic and functional characteristics of lymphoid cells infiltrating LMP tumors and invasive ovarian carcinomas. PMID- 8418078 TI - Florid mesothelial hyperplasia associated with ovarian tumors: a potential source of error in tumor diagnosis and staging. AB - Five ovarian tumors were associated with florid mesothelial hyperplasia that in four cases created initial problems in histologic classification and staging of the tumors. In two cases, small nests, cords, and gland-like arrangements of atypical mesothelial cells within the cyst wall of a serous or mullerian mucinous tumor of borderline malignancy were initially considered to possibly represent, or were misdiagnosed as, foci of stromal invasion, lymphatic invasion, or both. In the three other cases--an endometrioid adenocarcinoma, a struma ovarii, and a yolk sac tumor--papillary and sheet-like proliferations of mesothelial cells involved the extraovarian pelvic peritoneum; in one of the cases, necrosis of the mesothelial cells was prominent. These foci of extra-ovarian mesothelial hyperplasia were interpreted as metastatic tumor intraoperatively (three cases) and on initial microscopic examination (two cases). Factors that may have predisposed to the mesothelial hyperplasia included large size of the primary tumor (two cases), the presence of metastatic tumor elsewhere in the pelvis (three cases), ascites (two cases), and coexistent endometriosis (two cases). Distinction of mesothelial proliferation from invasive or metastatic ovarian tumor is obviously important in correct classification, staging, and management of the tumor. Awareness of the occasional occurrence of florid mesothelial hyperplasia in patients with ovarian neoplasms and attention to the histologic, histochemical, and immunohistochemical features of the mesothelial proliferation should facilitate the correct diagnosis. PMID- 8418079 TI - Contralateral ovary in unilateral ovarian carcinoma: a search for preneoplastic lesions. AB - Contralateral ovaries from patients with unilateral ovarian carcinoma were examined and compared to ovaries from age-matched control patients without ovarian carcinoma. The number of inclusion cysts were increased in ovaries from patients with ovarian carcinoma compared to the controls (p < 0.01). In addition, inclusions from cases with ovarian carcinoma showed serous differentiation more frequently than the controls (p < 0.01; odds ratio = 10.0; 95% confidence interval = 1.2-78.1). An age-related increase in the number of inclusion cysts was seen in the study group but not in the control group. These findings support a role of surface inclusion cysts in the genesis of ovarian carcinoma. PMID- 8418080 TI - The distribution of type 1 chain ABH and related histo-blood group antigens in normal cycling human endometrium. AB - Cycling endometrial tissue was examined immunohistochemically for blood group antigens using a panel of monoclonal antibodies with specificity to type 1 chain carbohydrates. Staining was evaluated against the genetic background (ABO, Lewis, and saliva-secretor status) and the subphase of the menstrual cycle. Expression of type 1 chain antigens in most cases correlated with the genetic background; however, Le(a) and Le(b) antigens were in a few cases demonstrated in endometria with the erythrocyte Le(a-b-) phenotype and Le(b) antigen in erythrocyte Le(a + b ) endometria also. In addition, Le(b) antigen was preferentially expressed in endometria from blood group O individuals. Type 1 chain antigens were, in general, maximally demonstrated in the surface epithelium. Chain 1 A, H, ALe(b), Le(a), and disialosyl-Le(a) (dsLe(a)) determinants were demonstrated only sporadically in functionalis and basalis glands of cycling endometria. Staining for most type 1 chain antigens showed variations that were related to the histomorphology (layer and menstrual phase), and monosialosyl-Le(a) (msLe(a)) seemed to be a marker of secretory differentiation in the endometrium. Our results support the view that synthesis and expression of type 1 chain antigens in human endometrium is influenced by the genetic background and is modulated by the hormonal environment. PMID- 8418081 TI - Metastatic ovarian tumors in children: a report of 14 cases and review of the literature. AB - Fourteen cases of neoplasms metastatic to the ovaries in children 10 weeks to 15 years of age are reported. Eight tumors were neuroblastomas, 7 primary in the adrenal gland, and 1 primary in the posterior mediastinum. Three tumors were rhabdomyosarcomas primary in the ethmoid sinus, right occipital region, and left thigh, respectively. The final three tumors were a Ewing's sarcoma, a rhabdoid tumor, and a carcinoid tumor primary in the fibula, kidney, and lung, respectively. The ovarian involvement was an autopsy finding in nine of the patients. Three of the remaining five patients presented clinically with manifestations suggesting a primary ovarian tumor, and the final two patients, who had known extraovarian primary tumors, had symptomatic ovarian masses discovered during life. In one case of neuroblastoma and the case of rhabdoid tumor, the ovarian metastases were initially misinterpreted pathologically as primary ovarian cancers; the primary renal tumor was not discovered until autopsy in the latter case. The ovarian tumors were bilateral in 8 of the 14 cases. Ovarian enlargement was present in 10 cases. Our experience and that in the literature indicates that the childhood tumor that spreads to the ovary most frequently is the neuroblastoma and that rhabdomyosarcoma is the most common sarcoma of childhood that spreads to the ovary. The clinical features and the frequent bilaterality of ovarian metastatic tumors are helpful diagnostic features in many cases, but when the ovarian tumor is the presenting manifestation of the disease, is unilateral, or both, differentiation from various primary ovarian tumors may be difficult. PMID- 8418082 TI - Choriocarcinoma in a term placenta: pathologic diagnosis of tumor in an asymptomatic patient with metastatic disease. AB - A clinically unsuspected choriocarcinoma arose in a term placenta. The patient was hospitalized for 3 weeks antepartum with preterm labor. She was delivered of a normal female infant at 36 weeks' gestation. Mother and infant were discharged after 3 days. The placenta, 465 g, contained multiple white nodules, a single infarct, and retroplacental hemorrhage. By microscopy, there was multifocal choriocarcinoma. No villous stromal invasion was found. The patient was readmitted 6 days postpartum with a serum beta human chorionic gonadotropin (hCG) value of 741,640 mIU/ml and lung metastases. She received 9 courses of multiagent chemotherapy and is currently free of tumor 1 year after diagnosis. The infant, 46,XX, remained clinically free of tumor although she had a serum beta hCG of 37 mIU/ml at 1 week of life. This placenta provides evidence of choriocarcinoma developing directly from mature placental villi in a chromosomally normal gestation and depicts a situation wherein choriocarcinoma is diagnosed correctly in the presence of villous tissue. It also demonstrates the utility of placental examination in clinically abnormal gestations. PMID- 8418083 TI - Papilloma of Bartholin's gland duct cyst: first report of a case. AB - A 50-year-old woman underwent excision of a vaginal cyst. Its distal end lay in the submucosal tissue of the posterior lateral aspect of the vulvar vestibule near the orifice of Bartholin's gland. The caudal wall of the cyst contained a papilloma with an epithelial lining which consisted of columnar and stratified polygonal cells resembling squamous and transitional epithelium. The histochemical attributes of the papillary epithelium were homologous to those of Bartholin's gland. Immunohistochemical studies supported glandular and squamous differentiation. Evolution from a dysontogenetic remnant of Mullerian origin cannot be excluded with certainty, but location and histopathology were consistent with origin from Bartholin's gland duct. The presence of mucin and the absence of a smooth muscle investment exclude derivation from Gartner's duct. Cysts of Bartholin's gland are common, but solid benign tumors are rare. We have been unable to find a report of a papilloma of either vulva or vagina with features similar to those in our patient. PMID- 8418084 TI - Can monoclonal antibody staining by 323/A3 and Ca1 of benign breast biopsies predict the development of breast cancer? AB - Monoclonal antibodies (mAb) raised to Ca1 and 323/A3 proteins have been suggested as markers of the premalignant breast. To establish the validity of this hypothesis the immunohistochemical staining patterns of these mAb have been studied in two different groups of patients with benign breast disease. One group comprised patients with benign biopsies who later went on to develop breast cancer. The second group were patients with benign breast disease who have not developed breast cancer. The immunohistochemical staining patterns of the two groups were compared. The staining patterns between the groups, when ducto lobular, apocrine metaplasia and fibroadenomatous tissue were analysed, was similar. This occurred both for Ca1 and 323/A3 mAb. We conclude that the antigens identified by these mAb will not enable prediction of cancer development. PMID- 8418085 TI - Co-expression of CD4 and CD8 associated with elevated interleukin-4 in a cord T cell line derived by co-cultivating normal cord leukocytes and an HTLV-II producing simian leukocyte cell line (Si-IIA). AB - A new interleukin-2(IL-2)-dependent T cell line, designated CS-IIA, was established by co-cultivating normal human cord leukocytes and a lethally X irradiated HTLV-II-producing simian leukocyte cell line (Si-IIA). CS-IIA showed CD4 dominance during the early culture. However, after addition of IL-2, CS-IIA predominantly co-expressed CD4 and CD8 (69.5%) and also expressed the surface markers CD1-, CD3+, CD19-, CD25+ and HLA-DR+. A significantly elevated level of IL-4 (1697 pg/ml) was observed in the culture supernatant from CS-IIA. In addition, the conversion of phenotype from some CD4+CD8+ cells to CD4+CD8- was demonstrated by the neutralization assay using anti-IL-4 antibody. CS-IIA had a normal human karyotype and was free from Epstein-Barr virus nuclear antigen and immunoreactive with sera of HTLV-I- or HTLV-II-infected patients and anti-HTLV-1, p19 or p24 mAb. The provirus genome of HTLV-II was detected in this cell line by the polymerase chain reaction combined with a digoxigenin-enzyme-linked immunosorbent assay. However, electron microscopy of CS-IIA cells revealed no C type virus particles in the extracellular space. These results indicate that HTLV II can be transmitted from an HTLV-II-infected simian leukocyte cell line to human cord T lymphocytes and suggest that co-expression of CD4 and CD8 on T cells may be induced by the high level of IL-4, which can mediate CD8 induction on CD4+ T cell clones. PMID- 8418086 TI - Antitumour activity of miltefosine alone and after combination with platinum complexes on MXT mouse mammary carcinoma models. AB - Miltefosine, an alkylphosphocholine structurally related to alkyllysophospholipids showed highly selective antitumour activity against the hormone-sensitive variant of the s.c. transplantable MXT mouse mammary adenocarcinoma, the ovary-dependent MXT (M3.2), whereas it was inactive against the hormone-insensitive MXT (M3.2) OVEX variant. A dose of 32 mg/kg miltefosine p.o. daily for 5 weeks was well tolerated. Histopathological evaluation gave no signs of gastroenteral toxicity. After therapy the microarchitecture of the MXT (M3.2) tumours changed from that of a moderately differentiated adenocarcinoma to that of an anaplastic mammary carcinoma. A dose of 16 mg/kg miltefosine p.o. daily, though in effective per se, enhanced the antitumour activity of suboptimal i.p. doses of cisplatin and the hormone-like platinum analogue meso-1,2-bis(2,6 dichloro-4-hydroxyphenyl) ethylenediamine]dichloroplatinum(II). Furthermore, it was shown, that miltefosine exhibited no (anti)hormonal properties. However, the mechanism of action of miltefosine remains unclear. PMID- 8418087 TI - Evaluation of the antitumour activity of coumarin in prostate cancer models. AB - Responses to coumarin have been reported for patients suffering from malignant melanoma, metastatic renal carcinoma and, recently, advanced prostate cancer. These data together with some experimental evidence for antiprostatic effect prompted us to study the activity of coumarin in various prostate tumour models and evaluate the endocrine properties of this drug. In rats no antiandrogenic activity was found. The growth of Noble Nb-R prostate tumours of the rat was strongly inhibited by coumarin (40 mg/kg; administered three times per week), whereas the hormonally more sensitive Dunning R3327-G rat prostate carcinoma did not respond to coumarin (40 mg) even when the drug was administered daily. Coumarin was also shown to possess antimetastatic activity in a Dunning R3327 MatLu tumour model. In this model small pieces of the hormone-independent tumour were implanted into the ear of the animal and later resected to mimic the clinical situation where primary tumours have been removed. The number of lung metastases was reduced significantly by 40%-50% following the administration of coumarin (40 mg daily). PMID- 8418088 TI - Effect of N-methyl-N'-nitro-N-nitrosoguanidine on carbohydrate profiles of non metaplastic rat gastric mucosa. AB - The effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on the mucin phenotype of non-metaplastic gastric mucosa in the rat was studied histochemically. Animals were exposed to MNNG in drinking water (83 mg/l) for 12 weeks. Carcinogen treatment was then discontinued and the animals (27 in the treatment group and 25 in the control group) were examined after another 44 weeks. Glycosylation was analysed with histochemical stains for sialomucins and sulphomucins and with peroxidase-conjugated lectins (GS-II, SBA, DBA, UEA-I, and WGA). Sialo- and sulphomucins remained quantitatively unchanged, only a slight increase of acid mucins in the antral glands was observed. The analysis of the lectin binding patterns, however, revealed a significant increase for WGA-binding glycoproteins in the surface mucous cells and gastric pits, while DBA binding was significantly decreased (P < 0.05). GS-II lectin bound specifically to the proliferative compartment in the gastric fundus, consisting of mucous neck cells, and was significantly increased after MNNG treatment. No specific alterations were detected in lectin binding to parietal or chief cells. It is concluded, therefore, that treatment of gastric mucosa with MNNG alters the glycoprotein metabolism before intestinal metaplasia can be observed. PMID- 8418089 TI - Impact of chemotherapy on collagen metabolism: a study of serum PIIINP (aminoterminal propeptide of type III procollagen) in advanced sarcomas. AB - We have previously shown that the serum aminoterminal propeptide of type III procollagen (PIIINP) is a prognostic factor for survival in localised soft-tissue sarcomas, and that elevated values are frequent in metastatic disease. In the present study PIINP is analysed during chemotherapy in 26 patients with advanced sarcomas. Non-responders had a significantly higher pretreatment level of PIIINP than responders (P = 0.05), when only patients with no recent therapeutic interventions were studied. However, during chemotherapy PIIINP followed the clinical course of the malignant disease in only a minority of patients. Patients with recent surgery or recently completed chemotherapy had an increased pretreatment PIIINP value (P = 0.03). In these patients PIIINP declined during chemotherapy irrespective of tumour response. A pretreatment PIIINP level within the reference range tended to increase with time irrespective of response. Moreover, the values taken during a chemotherapy infusion were significantly higher than those immediately preceding the corresponding cycle (P = 0.001). Our results suggest that pretreatment PIIINP is of value as a prognostic factor for chemotherapy response in patients with advanced sarcomas. During chemotherapy PIIINP is of minor importance in monitoring response because of the influence of chemotherapy and other therapeutic interventions on the level of PIIINP. PMID- 8418090 TI - Descriptive epidemiology of gall-bladder cancer in Europe. AB - Trends in mortality from cancer of the gall-bladder and bile ducts over the period 1965-1989 were analysed for 25 European countries on the basis of official death certifications from the World Health Organization databank. A high mortality area--i.e. with overall death certification rates, world standard, around or over 2/100,000 men and 4/100,000 women in 1985-1989--was identified in Germany and the surrounding central European countries (Austria, Czechoslovakia, Hungary and Poland). The highest rates were in Hungary (3.9/100,000 men and 7.4/100,000 women). During the two decades considered, rates increased in Czechoslovakia and Hungary, remained stable in Poland and declined in Austria and Germany. Intermediate-mortality areas included Scandinavian countries (except Norway) and Switzerland: their rates in the late 1980s were between 1.5 and 2.5/100,000 men and between 2.2 and 4.2/100,000 women. Mortality increased in Finland and Sweden, declined in the Netherlands and Switzerland, and did not change consistently in Denmark. Low-mortality countries (i.e. with rates in 1985 1989 below 2.0/100,000 men and 2.5/100,000 women) included Belgium, France, Britain, Ireland, Norway, Bulgaria and Mediterranean countries. Over the last two decades, certification rates declined in Bulgaria and Great Britain, but increased in all other countries. The ratio between the countries with the highest and lowest gall-bladder cancer mortality rates declined from 21 to 12 in women, although they remained stable around 10 for men. The pattern was similar when analysis was restricted to truncated rates from patients aged between 35 and 64 years. These trends, and particularly the exceedingly high rates in central Europe, the low rates in Mediterranean countries and the low and declining rates in Britain and Ireland are discussed in terms of known (cholelithiasis) or potential (dietary) factors in gall-bladder cancer aetiology, and of trends in cholecystectomy rates. PMID- 8418092 TI - Second primary malignancies following gynecological tumours in Saarland, Germany, 1968-1987. AB - The occurrence of second primary malignancies was assessed among 2922 women with cervical cancer, 2721 women with endometrial cancer and 1745 women with malignant neoplasms of the ovaries, who were notified to the cancer registry of Saarland, Germany, between 1968 and 1987. Analyses are presented for second primary malignancies at any site and at the most important single sites. Among women with cervical cancer, 98 second primary malignancies were observed, slightly fewer than the 99.3 cases that would have been expected on the basis of the incidence rates of the general population (standardized incidence ratio, SIR = 0.99). More second malignancies were observed than expected in the urinary bladder (SIR = 3.51) while the opposite applied to second neoplasms in the breast (SIR = 0.56). Among women with endometrial and ovarian cancer, the numbers of observed cases slightly exceeded the expected numbers for second primary neoplasms at any site (SIR = 1.19 and 1.41 respectively). More second primary neoplasms were observed than expected in the colon, breast, urinary bladder and kidneys among these women. We discuss our results, which are in agreement with findings among North European and North American populations, from clinical and etiological points of view. PMID- 8418091 TI - Analysis of mutant P53 protein in osteosarcomas and other malignant and benign lesions of bone. AB - Alterations of tumour suppressor genes are considered crucial steps in the development of human cancers. Expressions of p53 protein, a product of the tumour suppressor gene altered most commonly in human cancers examined so far, were investigated immunohistochemically in 18 osteosarcomas and 40 other malignant and benign lesions of bone. A monoclonal antibody clone PAb240, which recognizes a common conformational epitope of mutant p53 proteins, stained nuclei of tumour cells in 12 of 18 osteosarcomas (67%). Six tumours (33%) particularly showed positive immunoreactions in more than half of the tumour cells. PAb240 also stained tumour cells in a small number of other malignant bone tumours, such as malignant fibrous histiocytoma, chondrosarcoma, and Ewing's sarcomas. Furthermore, a small number of cells of giant-cell tumours were positively stained. In contrast, PAb240 was completely negative in 21 benign bone tumours and reactive lesions examined. Another monoclonal antibody clone PAb1801, which reacts with both wild- and mutant-type p53 protein, reacted in nuclei of tumour cells of 7 osteosarcomas (39%). Most of those also reacted with PAb240. PAb1801 was expressed much more frequently in other malignant bone tumours and giant-cell tumours. In addition, PAb1801 showed intranuclear positive reactions in tumour cells of a benign chondroblastoma, and reactive cells such as actively proliferating preosteoblasts in a myositis ossificans and osteoclast-like giant cells in a giant-cell tumour. The immunoelectron-microscopic observation that p53 protein was localized in euchromatic areas of nuclei of osteosarcoma cells supported the specificity of immunoreaction for p53 protein, indicating an active role of p53 protein in the regulation of DNA synthesis and transcription. These findings suggest that point mutation of the p53 gene is frequently involved in the development of osteosarcomas. PAb240 may be a useful tool not only in screening point mutations of the p53 gene in osteosarcomas but also in the differential diagnosis between osteosarcomas and reactive bone-forming lesions. Expressions of mutant p53 protein were not correlated with any clinical or pathological factors examined, although the results should be confirmed in studies of a large number of osteosarcomas. PMID- 8418093 TI - Who pays for continuing education in nursing? AB - This study examines the determinants of employer support for attendance at nursing continuing education (CE) offerings. The experience of 1,171 nurses was examined in relation to their age, educational background, present job characteristics, and annual income. Position level and income were positively related to the extent of employer support. The findings were consistent with the view that employers support attendance at CE offerings based on professional status rather than the needs of nurses. PMID- 8418094 TI - Retention of continuing education participants. AB - The Midwest Alliance in Nursing (MAIN) used a multidirectional approach to recruit long-term care nurses to a three-year, federally funded continuing education (CE) program in geriatric nursing. Program staff used a project logo, frequent mailings, bimonthly telephone calls, a series of diverse educational offerings, and scattered social meetings to strengthen participants' identification with the project's purpose and activities. Nevertheless, several nurse dyads failed to complete the program, citing excessive workload and lack of agency support as principal reasons for withdrawal. Program staff concluded that, without strong and continuing support from their employer, even highly motivated nurses will not maintain commitment to an extended-time CE program. PMID- 8418095 TI - A descriptive study of the CE needs of nurses serving adolescents in the community. AB - The purpose of this descriptive study was to develop a two-pronged community education database to improve the quality of response to professionals' self assessed educational needs. The database was designed to include both the self assessed level of the respondents' learning needs and their perception of the importance of mastery of specific content in reaching targeted outcomes for the adolescents they serve. Sixty-one percent of the 425 surveys mailed to professionals from central and southeastern Ohio were analyzed. Responses were weighted and a scattergraph of the mean scores for each topic was plotted. Implications for nursing continuing education (CE) are discussed. PMID- 8418096 TI - Teaching family/significant other nursing. AB - A family/significant other nursing education program for staff nurses can benefit staff, patients, families, and the organization. Recent Joint Commission on Accreditation of Healthcare Organizations (JCAHO) focus survey processes attempt to evaluate how nursing departments operationalize the JCAHO nursing standards. Nurses must be able to articulate and document their organization's policies and procedures regarding the nursing care standards, including those standards that address family and significant other needs. PMID- 8418097 TI - State and association/certifying boards CE requirements. PMID- 8418098 TI - Career ladder program. AB - In June 1990, Santa Rosa Children's Hospital instituted a multitrack career ladder program for registered nurses, in response to the issues of retention of expert nursing staff and development of professional nursing practice. The program incorporates clinical, education, and management tracks, providing vertical and lateral mobility to the registered nurse. Performance evaluation criteria complement the promotional process, and the registered nurse has a menu of requirements to fulfill each year upon promotion to or maintenance of career ladder level. Through incorporation of a graduated salary system, the program has been a retention bonus for registered nurses at Santa Rosa Children's Hospital. In addition, the career ladder program has been very successful in promoting professional nursing practice within the hospital. PMID- 8418099 TI - "The Princess and the Chemo Spill"--a policy magically turned into a fairy tale. PMID- 8418100 TI - Interactive television presentation style and teaching materials. AB - Special teaching techniques and presentation style are required for an effective telecourse. This article describes modification of a traditional graduate nursing course for televised delivery to remote students. Teacher appearance and behaviors as well as student activities for telecourses are proposed. PMID- 8418102 TI - Macrotrends in nursing practice: what's in the pipeline. PMID- 8418101 TI - Juggling performance checklist. AB - Teaching psychomotor skills to nurses presents challenges to nurse preceptors. The Juggling Performance Checklist, used by The Presbyterian Hospital in New York City, presents a humorous way of teaching nurses how to teach psychomotor skills. Humor can have both a physical and psychological benefit. Please note the directions on the performance checklist--washing your hands is optional; having fun is not! PMID- 8418103 TI - The role of phospholipase A2 in interleukin-1 alpha-mediated inhibition of mineralization of the osteoid formed by fetal rat calvaria cells in vitro. AB - Interleukin-1 (IL-1) may be an important mediator of bone remodeling, since it is a potent stimulator of bone resorption and has biphasic effects on bone formation. Continuous exposure of fetal rat calvaria (RC) cells to IL-1 alpha or IL-1 beta results in a dose-dependent inhibition of both bone nodule formation and mineralization of the organic matrix. In this study, the effects of recombinant human IL-1 alpha on the mineralization process were examined by the addition of IL-1 alpha late in the culture period, after osteoid nodules had formed and when they were induced to mineralize by the addition of organic phosphate. By means of a quantitative 45calcium radiolabeling assay, it was shown that short-duration exposures of fully-formed bone nodules to IL-1 alpha also inhibited mineralization, and that the duration of treatment directly correlated with the degree of inhibition. Because our earlier studies had demonstrated that IL-1 stimulated the release of PLA2 and PGE2 from RC cells, the effects of PLA2 and of inhibition of PGE2 synthesis on mineralization were investigated. Exogenous Naja naja group I PLA2 had little effect on the mineralization of bone nodules; however, Crotalus adamanteus group II PLA2 inhibited mineralization at concentrations similar to those found in the media from IL-1 alpha-treated cultures. Although PLA2 is thought to stimulate PGE2 synthesis by releasing arachidonic acid from membrane phospholipids, PGE2 release by RC cells accounted for only part of the IL-1 alpha-mediated inhibition, suggesting the presence of other mechanisms of exogenous PLA2 action in inhibiting mineralization.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418104 TI - Distribution and isolation frequency of eight streptococcal species in saliva from predentate and dentate children and adults. AB - The isolation frequency and distribution of eight recently defined streptococcal species have been investigated in the saliva of adults and that of both predentate and dentate children. The effects of frequency of sugar intake and tooth presence on the distribution of streptococcal species were also analyzed. Saliva samples were collected from 121 subjects divided into three study groups: (a) 56 predentate children (predentate group), (b) 37 dentate children (dentate group), and (c) 28 adults (adult group). Up to 17 biochemical and enzymatic tests were used to categorize streptococcal isolates into S. mitis, S. oralis, S. salivarius, S. anginosus, S. sanguis, S. vestibularis, S. mutans, and S. gordonii. The mean total and streptococcal salivary colony-forming units (CFU) were lowest in the predentate group and highest in the adult group. Streptococci were found in all the study subjects, and there was no obvious relationship between the total or streptococcal CFU and the number of teeth or the frequency of sugar intake. There was a wide variation in the isolation frequency of streptococcal species in the three study groups. S. mitis, S. oralis, and S. salivarius were the most frequent species isolated, and together they comprised 83% of the total streptococcal isolates. In contrast to studies using older classifications, S. sanguis was a minor species in the saliva though found more often in adults than in children (p < 0.04). S. anginosus was a minor species found in about 10% of adults and children. S. gordonii was detected rarely and only in dentate subjects. S. mutans was detected only in dentate subjects, significantly greater in adults (57.1%) than in children (5.4%, p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418105 TI - Initial acquisition of mutans streptococci by infants: evidence for a discrete window of infectivity. AB - Oral bacterial levels of 46 mother-child pairs were monitored from infant birth up to five years of age so that the acquisition of mutans streptococci (MS) by children could be studied. The initial acquisition of MS occurred in 38 children at the median age of 26 months during a discrete period we designated as the "window of infectivity". MS remained undetected in eight children (17%) until the end of the study period (median age of 56 mo). The levels of both MS and lactobacilli in saliva of mothers of children with and without MS were not significantly different. Comparisons between a caries-active cohort colonized by MS (nine of 38) and children without detectable MS revealed similar histories in terms of antibiotic usage, gestational age, and birth weight. Interestingly, half of the children between the ages of one and two years who were not colonized by MS were attended by caretakers other than the mother, while all of the caries active children during this same time period were cared for by their mothers; the difference was statistically significant. Here we report for the first time that MS is acquired by infants during a defined period in the ontogeny of a child. Support for the notion of a discrete window of infectivity comes from other sources, including animal models. PMID- 8418106 TI - Comparison of rates of enamel synthesis in impeded and unimpeded rat incisors. AB - Periodic intubations of rats with solutions of fluoride (F) lead to the appearance of bands of disrupted pigmentation in continuously erupting incisors. Distances between fluorotic bands reflect time intervals between intubations. In this experiment, the periodicity of fluorotic banding was used for estimation of the rate of enamel synthesis in impeded and unimpeded rat incisors. Rats kept on a low-F diet and distilled water were intubated two or four times per week with 2 mg NaF/150 g body weight. In a group of rats, one of the mandibular incisors was cut at the gingival margin after two weeks, and intubations were continued for an additional two weeks. In another group of F-intubated rats, incisors were cut or notched at the gingival margin twice, six days apart. Control rats either received the same periodic F intubations or were maintained on the low-F diet without intubation. Measurements of spacing between fluorotic bands were identical in impeded and unimpeded teeth, even though the latter erupted at a faster rate. In an unimpeded mandibular incisors, there was a significant elongation of the secretory zone and a shortening of the pigmentation zone, resulting in reduced pigmentation intensity of the erupted portions of the teeth. The results show that the rate of enamel synthesis is independent of the eruption rate. PMID- 8418107 TI - Effect of jaw opening on masticatory muscle EMG-force characteristics. AB - This study was designed to evaluate the way in which changes in vertical jaw opening affected the relative contributions of various masticatory muscles to bite force production. EMG activity was recorded simultaneously from the masseter, and anterior, middle, and posterior temporalis muscles, during controlled isometric biting at different force levels and vertical jaw openings. EMG-force characteristics were compared between muscles and bite openings. All but the posterior temporalis muscle displayed significant increases in muscle activity with increased bite force production; the masseter muscle demonstrated the largest activity increments. Statistically significant changes in muscle function due to jaw opening were demonstrated only for the masseter muscle, though similar trends were observed for the anterior and middle temporalis muscles. Minimum increases in muscle activity associated with increases in bite force occurred between 9 and 11 mm of opening, measured at the first molar, for all three muscle groups. The results of this study suggest that changes in masticatory muscle length resulting from vertical jaw opening cause alterations in muscle contractile properties, but the relative contributions of various masticatory muscles toward bite force production may also be affected by biomechanical factors and neural control adaptations. PMID- 8418108 TI - Influence of shearing action of food on contact stress and subsequent wear of stress-bearing composites. AB - The influence of sliding action of the antagonist on occlusal three-body wear of composites and an amalgam was investigated in vitro by gradual change in the distance between the opposing substrates. When the distance was decreased from 10 microns to approximately 3 microns, wear increased significantly by a factor of two to three and was exclusively of erosive nature. At a slurry-film thickness of approximately 1 micron, direct contacts between the antagonist and protruding composite filler particles started to occur. This consequently slowed the erosive wear. Ultimately, direct contact phenomena predominated, decreasing the wear rate of the various materials to different degrees. Loss of material due to subsurface fatigue could not be demonstrated with a contact pressure of 45 MPa at which the experiments were performed. From this study, it can be concluded that minor alterations of the food-film thickness at the contact areas result in considerable changes in wear rates and wear-rate ranking of composite materials, which may partly explain inconsistencies among clinical trials. PMID- 8418109 TI - Relationships between the temporomandibular joint and the middle ear in human fetuses. AB - A study was conducted, on 30 human fetuses, of the structures passing through the tympanosquamosal fissure. The tympanosquamosal fissure lies between the middle ear and the temporomandibular region. Meckel's cartilage passes through the tympanosquamosal fissure and continues on into the middle ear with the cartilaginous anlage of the malleus. A tract of fibrous tissue arises from the mesenchyme, located cranial and lateral to Meckel's cartilage, that enters from the posterior area of the temporomandibular joint disc to the middle ear through the tympanosquamosal fissure, and attaches onto the area of continuity of Meckel's cartilage with the malleus. Transformation of Meckel's cartilage into the sphenomandibular ligament and anterior ligament of the malleus determines their continuity through the tympanosquamosal fissure. The posterior fibers of the temporomandibular joint disc giving rise to the discomalleolar ligament insert into the anterior ligament of the malleus. PMID- 8418110 TI - The effects of etching, micro-abrasion, and bleaching on surface enamel. AB - Conservative techniques for improving the appearance of discolored teeth have become popular in the past decade. These include: in-office bleaching with 30% hydrogen peroxide, which is applied on etched enamel with a gauze pad and then exposed to a bleaching light; home bleaching with a mild form of peroxide, such as 10% carbamide peroxide, which is applied on the tooth surface with custom-made mouthguards; and enamel micro-abrasion with 18% hydrochloric acid, which is applied in a pumice slurry. In this study, the in-office bleaching and enamel micro-abrasion techniques were performed on extracted teeth for investigation of their microscopic effects on the surface enamel. Specimens treated only with 37% phosphoric acid showed an enamel loss of 5.7 +/- 1.8 microns. The specimens treated with 37% phosphoric acid followed by 30% hydrogen peroxide showed enamel loss of 5.3 +/- 1.6 microns; this loss was probably not caused by the hydrogen peroxide, but rather by the etching with 37% phosphoric acid which preceded the hydrogen peroxide application. A direct application of 18% hydrochloric acid for 100 s resulted in a loss of 100 +/- 47 microns. The extent of enamel loss was much greater when the 18% hydrochloric acid was applied in a pumice slurry for the same period of time (360 +/- 130 microns), and the effect was time-dependent. Thus, the pumice and rotary prophy cup used in conjunction with the 18% hydrochloric acid contributed markedly to the loss of surface enamel, enhancing the non-selective stain-removing action of the hydrochloric acid. Therefore, the hydrochloric acid-pumice technique must be used clinically with caution. PMID- 8418111 TI - The determination of ionic bonding interactions of N-phenyl glycine and N-(2 hydroxy-3-methacryloxypropyl)-N-phenyl glycine as measured by carbon-13 NMR analysis. AB - The purpose of the present investigation was to determine whether high-resolution carbon-13 nuclear magnetic resonance could be utilized for detection of ionic bonding interactions of NPG and NPG-GMA with selected inorganic cations. The C1 carbonyl carbon of NPG and NPG-GMA were labeled with 13C to help magnify the spectral response signal. The labeled monomers were allowed to react with Ca(OH)2 and LiOH. The C-13 carbonyl resonance was observed by solution- and solid-state NMR. The interaction of the cations in solution and on a powered glass support resulted in a downfield chemical shift of about 2-5 ppm, indicating the presence of a chemical interaction between the monomer and the cations. The reaction of the monomers with the solid substrates of hydroxyapatite, and bovine dentin did not result in a significant downfield shift in the carbonyl resonance. The results provide evidence that high-resolution nuclear magnetic resonance can be used for evaluation of chemical bonding interactions of dental adhesives with inorganic ions. PMID- 8418112 TI - A quantitative study of calcium binding and aggregation in selected oral bacteria. AB - By means of micro-equilibrium dialysis, calcium binding capacities and affinities were measured in three different oral bacteria, and the effects of extracellular polysaccharide, pH, and aggregation were investigated. Binding capacities of 31.0 +/- 2.1 (C. matruchotii), 34.7 +/- 3.7 (S. sanguis), and 41.5 +/- 5.4 (S. downei) mumol calcium/g wet weight of cells were found at pH 7.0, falling to 21.4 +/- 0.8 mumol calcium/g wet wt. cells at pH 5.0 for S. downei. Dissociation constants were found to vary between 0.78 +/- 0.24 and 1.77 +/- 0.30 mmol/L (at pH 7.0, depending on species), and between 0.62 +/- 0.04 and 1.77 +/- 0.30 (in the pH range 5.0 to 7.0, for S. downei only). Examination suggested that at pH 7.0 calcium-facilitated bacterial association occurs in the streptococci with calcium uptake curves analogous with those of positively cooperative systems. Desorption of calcium from aggregated S. downei suggested that the mechanism of desorption differed from that of uptake. This may be an important factor in plaque formation and in the binding of cells to the surface of formed plaque. Plaque calcium forms a reservoir, readily released by a pH drop, which may increase plaque fluid saturation and reduce demineralization. PMID- 8418113 TI - Elective pulpectomy (the need to recycle some old information) PMID- 8418114 TI - Risk indicators and risk markers for periodontal disease experience in older adults living independently in Ontario, Canada. AB - This study examined risk indicators and risk markers for periodontal disease experience in 624 adults aged 50 years and over living independently in four communities in Ontario, Canada. The data were collected as part of the baseline phase of a longitudinal study of the oral health and treatment needs of this population. Periodontal disease experience was assessed in terms of attachment loss, measured at two sites on each remaining tooth. Bivariate and multivariate analyses were used to examine the relationship between a number of sociodemographic, general health, psychosocial, and oral health variables and three indicators of periodontal disease experience. These were: mean attachment loss, the proportion of sites examined with loss of 2 mm or more, and the probability of the subjects having severe disease, arbitrarily defined as a mean attachment loss in the upper 20th percentile of the distribution. Mean attachment loss was 2.95 mm (SD = 1.41 mm), and 76.6% of sites examined had loss of 2 mm or more. In bivariate analyses, the most consistent predictors of periodontal disease experience were: age, education, income, smoking, dental visiting, the number of remaining teeth, the number of decayed coronal surfaces, and the number of decayed root surfaces. In multivariate analyses, age, education, current smoking status, and the number of teeth had the most consistent independent effects. These data confirm the results of recent US studies indicating that periodontal disease experience is influenced by social and behavioral factors. PMID- 8418115 TI - Influenza A virus-specific cytotoxic T lymphocyte activity declines with advancing age. AB - OBJECTIVE: To investigate whether influenza A-specific cytotoxic T lymphocyte (CTL) activity is reduced in elderly compared with younger adults. DESIGN: Case series comparing outcomes in young and elderly cohorts. SETTING: Saint Louis University Division of Geriatric Medicine. PARTICIPANTS: Healthy adult outpatients and staff members aged < 50 (young) or > or = 65 (elderly) years. METHODS: Peripheral blood mononuclear cells were assayed for CTL activity by a 51chromium release assay following 1 week of in vitro stimulation with influenza A/PR/8/34 (H1N1) virus. MEASUREMENTS: Percent specific lysis of autologous and allogeneic influenza virus-infected target cells. MAIN RESULTS: Specific lysis of autologous A/PR-infected targets was significantly lower in elderly compared to young subjects (P < 0.01), and exceeded 10% in a significantly lower proportion of elderly compared with younger subjects (P < 0.05), but was not influenced by a history of vaccination within the preceding 12 months. Cytotoxic effectors were class I human leukocyte antigen (HLA)-restricted and displayed heterosubtypic cross-reactivity but were unable to lyse influenza B-infected targets. CONCLUSIONS: These results demonstrate an age-related decline of influenza A virus-specific CTL activity and suggest that CTL responses to inactivated virus vaccine are of short duration. PMID- 8418116 TI - A comparative study of monoclonal gammopathies and immunoglobulin levels in Japanese and United States elderly. AB - OBJECTIVE: To define the prevalence of monoclonal immunoglobulin (Ig) proteins and quantitative serum immunoglobulin levels in elderly Japanese in comparison with elderly Caucasians as possible factors related to the reported lower incidence of multiple myeloma in elderly Japanese than in elderly Caucasians. DESIGN: Survey study SETTING: Community Center in Yokohama, Japan and Retirement Community in the United States. PARTICIPANTS: Convenience sample of community dwelling elderly subjects (age 63-95) presenting for health screening examinations in each setting. Frozen serum samples were obtained from routine screening from 146 consecutive Japanese subjects and 111 US subjects. INTERVENTION: None MEASUREMENTS: Presence of monoclonal immunoglobulin protein determined by serum protein electrophoresis and immunofixation and quantitative Ig by laser nephelometry. RESULTS: Four (2.7%) of the Japanese cohort had monoclonal gammopathies compared with 11 (10%) of the American cohort. Two of the monoclonal gammopathies were IgG Kappa and two were IgG Lambda. No cases of multiple monoclonal gammopathy were identified in the Japanese group, compared with 25% of the monoclonal gammopathies in the American group. The mean quantitative serum IgG level for the Japanese group was 1,685 +/- 520 mg/dL versus 1,118 +/- 402 mg/dL for the American group; mean quantitative IgA levels were 283 +/- 116 mg/dL versus 226 +/- 116 mg/dL (P < 0.001). Albumin levels were normal in both populations, suggesting that there was not an increase in occult inflammatory disorders in the Japanese population. CONCLUSION: The low prevalence of monoclonal gammopathy in elderly Japanese is consistent with the reported lower frequency of multiple myeloma. The reason for the higher quantitative immunoglobulin levels in this population is unclear. Further cross-cultural investigation is warranted to explore the genetic influences on altered immune regulation with aging. PMID- 8418117 TI - Treadmill walking in old age may not reproduce the real life situation. AB - OBJECTIVE: In a preliminary study in our laboratory, healthy elderly people had a higher heart rate during treadmill walking than during corridor walking at the same speed. The objective of this study was to determine whether this initial observation, (1) persisted after repeated testing, (2) was present in younger adults, (3) was due to wearing a mouthpiece during treadmill walking, or (4) was due to a change in gait. DESIGN: A study of elderly and young volunteers undergoing repeated testing, with comparison of treadmill walking with corridor walking. SETTING: The Royal Free Hospital School of Medicine. PARTICIPANTS: Twelve healthy elderly (71-80 years) and 12 healthy young (21-37 years) volunteers. MAIN OUTCOME MEASURES: Heart rate (beats/min) and step rate (steps/min) during comfortable self-paced corridor walking and during treadmill walking at the same speed. MAIN RESULTS: The elderly subjects had higher heart rates during treadmill walking than during corridor walking at the same speed (mean difference = 6 beats/min, 95% Confidence Interval (CI) = 1 to 10). This difference increased (to a mean of 11 beats/min, 95% CI = 5 to 16) when a mouthpiece was worn on the treadmill. These differences persisted after repeated testing. The young subjects did not have higher heart rates on the treadmill, (with or without the mouthpiece). In both groups, step rate was lower (95% CI = 9 to -2, elderly; -5 to -2, young) during treadmill walking, corresponding to a 3% increase in stride length. CONCLUSION: The heart rate response to treadmill walking in healthy elderly people may be less representative of the "real life" situation than in younger adults. PMID- 8418118 TI - Transcranial Doppler assessment of the cerebral circulation during postprandial hypotension in the elderly. AB - OBJECTIVE: The aim of the present study was to evaluate whether alterations in postprandial hemodynamics in the elderly were associated with changes in cerebral perfusion assessed by transcranial Doppler ultrasonography. DESIGN: Time series, ie, post-intervention compared to pre-intervention with no-intervention controls. PARTICIPANTS: Ten elderly institutionalized subjects (4 women, 6 men, mean age 84.9 years). Three subjects had a history of syncope. SETTING: A 725-bed academic long-term care facility. INTERVENTION: A 400-kcal mixed meal. MEASUREMENTS: Heart rate, blood pressure, and blood flow velocity in the middle cerebral artery by transcranial Doppler recording, before the test meal and at 5-minute intervals for 60 minutes afterwards. RESULTS: Systolic, diastolic, and mean arterial blood pressure declined significantly from baseline between 30 and 55 minutes after the meal (P < 0.05, ANOVA); however, maximum and mean blood flow velocity did not change. The pulsatility index (end diastolic to peak systolic amplitude divided by mean velocity) increased significantly (P < 0.05, ANOVA) between 30 and 55 minutes after the meal, suggesting increased arteriolar resistance. There were no significant changes in blood pressure, blood flow velocity, and pulsatility index during a control study conducted with four subjects under identical conditions but without a meal. CONCLUSIONS: The results of this study suggest a small, unexpected increase in resistance of the intracranial circulation following a meal in elderly people with postprandial hypotension. Although the clinical significance of this finding is not known, the occurrence of postprandial arteriolar vasoconstriction may lead to cerebral ischemia during periods of marked blood pressure decline. PMID- 8418119 TI - Clinical features distinguishing large cohorts with possible AD, probable AD, and mixed dementia. AB - OBJECTIVE: To determine whether clinical features and rate of cognitive and functional decline differed in cohorts of possible AD (poAD), probable AD (prAD), and mixed dementia (MIX) patients. DESIGN: Cohort study with 1-year follow-up examination, comparing three groups of subjects. SETTING: Outpatient evaluation at nine California Alzheimer's Disease Diagnostic and Treatment Centers (ADDTC). PATIENTS: There were 1701 elderly patients who presented for evaluation of memory complaints. MEASUREMENTS: Historical, physical, and neurological variables for cross-sectional comparisons and 1-year rate of change on the Mini-Mental State Examination (MMSE), Blessed Information-Memory-Concentration test (BIMC), and Blessed Dementia Scale (BDS). RESULTS: Mean initial MMSE scores for poAD (n = 279), prAD (n = 928) and MIX (n = 430) were 17.9 (+/- 7.4), 13.9 (+/- 7.5), and 15.4 (+/- 7.1). Delusions and psychosis occurred in about one-third of each group, most often in those with moderate dementia (MMSE 11-20). PoAD were distinguished from prAD by significantly more alcohol abuse, physical health problems, and focal motor or sensory findings. MIX differed from AD alone by increased prevalence of cardiovascular disease, hypertension, stroke, TIA, and exposure to general anesthesia, and by a greater frequency on exam of depressed mood, focal motor or sensory findings, and gait disorder. All groups declined by about 2.8 points on the BIMC, 2.9 points on the MMSE, and 1.8 points on the BDS, a functional scale, over 1 year. Neither extrapyramidal signs nor psychosis predicted a more rapid rate of decline. CONCLUSIONS: Various features help to distinguish poAD, prAD, and MIX in a large cohort of patients, but do not predict the rate of progression. PMID- 8418120 TI - Validation of a measure of physical illness burden at autopsy: the Cumulative Illness Rating Scale. AB - OBJECTIVE: To further validate an objective measure of physical illness burden, the Cumulative Illness Rating Scale (CIRS). DESIGN: Survey with correlation of CIRS ratings made from physician interviews and review of medical records with post-mortem ratings made independently at tissue autopsy. SUBJECTS: Victims of completed suicide investigated by both psychological and tissue autopsy (n = 72). RESULTS: CIRS ratings made by examination of tissue at autopsy were highly predictive of analogous ratings based on historical data, accounting for 75% of the variance in CIRS scores. Taking autopsy findings as the gold standard of objective health assessment, historical ratings tend to underestimate physical illness at high levels of tissue pathology and to overestimate burden at lower levels. CONCLUSIONS: The CIRS score, when derived from all available sources of medical information, is a valid objective measure of physical illness burden and has broad applicability to research in geriatrics. PMID- 8418121 TI - Correction of massive vaginal prolapse in an older population: a four-year experience at a rural tertiary care center. AB - OBJECTIVE: To ascertain the success of complex reconstructive vaginal surgery in older women. DESIGN: Retrospective review of hospital and outpatient records. SETTING: Rural tertiary care referral center, Pennsylvania State University Hospital, Hershey, Pennsylvania. PATIENTS: Twenty-four patients referred for massive erosion of the vagina and/or complete procidentia. MEASUREMENTS: Symptoms and anatomic correction of patients' complaints. RESULTS: After surgery, 83 percent were asymptomatic without pelvic relaxation, 4 percent were asymptomatic with pelvic relaxation, 4 percent were symptomatic without pelvic relaxation, and 9 percent were symptomatic with relaxation. CONCLUSIONS: Older women can undergo major vaginal reconstructive surgery with relief of symptoms and restoration of vaginal depth and axis. PMID- 8418122 TI - Rate of progression and prognostic factors in Alzheimer's disease: a prospective study. AB - OBJECTIVE: To study rate of progression and correlates of cognitive and functional/behavioral deterioration in Alzheimer patients. DESIGN: A 1-year multicenter prospective study. SETTING: Outpatients and inpatients at geriatric institutions. PATIENTS: Fifty-six patients with a clinical diagnosis of Alzheimer's disease according to DSM-III criteria of Primary Degenerative Dementia. MAIN OUTCOME MEASURES: Blessed Dementia Scale (BDS) and Blessed Information-Memory-Concentration test (BIMC) measured at baseline, third, sixth, and twelfth month. RESULTS: The mean annual (+/- SD) rate of progression of our sample was 3.5 (+/- 3.7) points on the BDS and 2.6 (+/- 4.9) on the BIMC, with a wide range of variability. The level of cognitive impairment (BIMC score) at baseline predicted functional and behavioral deterioration: the better the initial BIMC score, the less the rate of negative change of BDS (r = 0.37, P = 0.006). Furthermore, the younger the patients at the disease onset, the faster the progression of cognitive impairment (r = 0.48, P = 0.0003), with men having a slower rate of progression (P = 0.004) than women. CONCLUSIONS: The present study confirms previous findings showing a wide individual variability in rate of progression of cognitive and functional/behavioral impairment as assessed by BIMC and BDS. The cognitive profile may predict the clinical evolution better than the functional and behavioral status, with patients with an earlier onset and women having a faster deterioration. PMID- 8418123 TI - A brief agitation rating scale (BARS) for nursing home elderly. AB - OBJECTIVE: To determine the effectiveness of a brief agitation rating scale (BARS) derived from the Cohen-Mansfield Agitation Inventory (CMAI). DESIGN: Test of reliability and validity of a new test. SETTING: The Lieberman Geriatric Health Centre, a skilled nursing facility. PARTICIPANTS: Forty members of the Lieberman Geriatric Health Centre nursing staff, 232 residents, and a sample of 40 randomly selected residents. MEASUREMENTS: Observational agitation ratings by registered nurses and certified nurses' assistants; item to total correlations for the CMAI on each of the three shifts; Pearson correlations between the 10 item BARS and the BEHAVE-AD and the BSSAD scales. RESULTS: Internal consistency reliability for the BARS was .74 (for the day shift), .82 (evening shift), and .80 (night shift), compared with .86, .91, and .87 for the full CMAI. Each of the 232 residents' scores on the BARS was correlated with a total score from the CMAI separately on each shift. Correlations were of .95, .94, and .95. Thus, across all shifts, the BARS accounted for approximately 90 percent of the variance of the total score of the CMAI. Concurrent validity of the BARS was supported by significant correlation with BEHAVE-AD and BSSD. CONCLUSIONS: The BARS represents a brief and effective mechanism to assess the presence and severity of physically aggressive, physically non-aggressive, and verbally agitated behaviors in elderly nursing home residents. PMID- 8418124 TI - A comparison of hand-held isometric strength measurement with isokinetic muscle strength measurement in the elderly. AB - OBJECTIVE: To compare hand-held isometric muscle strength measurement to an isokinetic muscle strength measurement in a healthy elderly population. DESIGN: Cross-sectional survey. SETTING: Retirement community in Southeastern Arizona. PARTICIPANTS: Thirty-two volunteers (16 men and 16 women) age 60 and over (mean age 70.3 years) who were free of musculoskeletal problems and did not have significant health problems. INTERVENTIONS: None. MEASUREMENTS: Isometric muscle strength was determined using a hand-held isometric strength device (Penny and Giles) for elbow flexion and extension and knee flexion and extension. Isokinetic muscle strength (average peak torque and average work per repetition) was measured on the same individuals using a Lido isokinetic dynamometer. RESULTS: Correlations between the strength measurement techniques were generally favorable with the lowest correlation being .72, (95 percent confidence interval .50-.86) and the highest being .85, (95 percent confidence interval .72-.93). However, there was substantial variability of hand-held strength reading at some levels of isokinetic strength. CONCLUSIONS: Hand-held dynamometry for strength measurement correlates strongly with measurement of strength using isokinetic dynamometry. Despite these high correlations, there is remaining variability of hand-held muscle strength readings when compared with isokinetic strength measurement at some levels of isokinetic muscle strength. Modification of the testing methodology or instrumentation used in this study is needed to improve the consistency of these measurements. PMID- 8418125 TI - Alcoholism and prescription drug abuse in the elderly: St. Louis University grand rounds. AB - This Grand Rounds will review the problem of alcoholism and prescription drug abuse in the elderly. Several case vignettes will be presented. The pharmacology of alcohol and potentially addictive prescription medications will be reviewed. The clinical presentation of and psychiatric symptoms associated with these disorders will be discussed. The process of addiction and issues regarding the clinical evaluation of the elderly addict will be discussed. The medical complications of these disorders will be reviewed, followed by a discussion of guidelines for the appropriate use of these drugs in the elderly. The grand rounds will conclude with a discussion of the treatment of patients with these disorders. PMID- 8418126 TI - When older adults face the chair-rise challenge. A study of chair height availability and height-modified chair-rise performance in the elderly. AB - OBJECTIVE: Define the range of community seating heights available for use by older adults; test whether raising chair height by small increments facilitates chair-rise performance; and heighten physician and furniture industry awareness of discrepancies that may exist between actual and acceptable chair heights for older adults. DESIGN: Phase 1: Survey of commercially available chair heights. Phase 2: Cross-sectional descriptive study of chair-rise ability. SETTING: Phase 1: Local furniture stores, physician offices, hospital waiting areas, and nursing homes. Phase 2: Postural Control Lab. PARTICIPANTS: Twenty-two volunteers (nursing home residents and community dwellers). EXCLUSION CRITERIA: inability to stand independently and inability to bear full weight on the lower extremities in the standing position. MEASUREMENTS: Chair rise success at six heights (17-22 inches), self-reported difficulty (visual analogue scale), change in minimum hip angle and maximum shoulder angle during rise, using motion analysis. RESULTS: Phase 1: Community chair heights ranged from 12 to 18 inches, with a mean of 16.3 in physician offices, 16.6 in nursing homes, 16.4 in hospitals, 17.3 in "kitchens" and 15 in "living rooms." Phase 2: As chair height increased from 17 to 22 inches, chair rise effort decreased, as shown by near doubling of percent successful rises, decline in mean self-reported difficulty score, increase in mean minimum hip angle, and decrease in mean maximum shoulder angle. CONCLUSIONS: Seating height may need to be more closely scrutinized in areas frequented by frail elders. Augmentation of seat height by small increments facilitates chair rise performance. PMID- 8418127 TI - Decisions about enteral tube feeding among the elderly. AB - OBJECTIVE: To determine choices about enteral tube feeding and factors associated with deciding to accept or forego this intervention in a group of ambulatory non demented older individuals. DESIGN: Descriptive survey. SETTING AND PARTICIPANTS: Thirty four volunteers from a senior adult day center and 34 volunteers from the residential care section of a multilevel long-term care institution, mean age 77.8. INTERVENTION: Structured interview using a hypothetical clinical vignette in simplified language, story-book format depicting an irreversibly and severely impaired state of health. MEASUREMENT: Choice of whether to accept or forego enteral tube feeding based on the clinical vignette. RESULTS: Thirty four (50%) decided to accept and 34 (50%) chose to forego enteral tube feeding in the situation presented in the vignette. No demographic, cognitive, or affective factors were associated with the decision. Presentation of the vignette and associated questions were not anxiety-provoking or upsetting to the vast majority of participants. CONCLUSION: A hypothetical clinical vignette depicting a state of severely impaired health resulted in 34 (50%) of 68 ambulatory non-demented older individuals deciding to accept enteral tube feeding. No factors we examined were strongly associated with the decision. The vignette and discussion were not anxiety-provoking when presented in the format used in this study. Advance directive discussions about enteral tube feeding and other health care decisions, using understandable hypothetical clinical vignettes that describe risks and benefits that might influence decisions, should be encouraged in the practice of geriatric medicine. PMID- 8418128 TI - The geriatric "medical and public health" imperative revisited. AB - Attempts to reduce the future demand for institutional care through community services are likely to have limited success. For this reason, health professionals must focus on preventing or ameliorating functional decline in older persons. To focus attention on this aspect of the geriatric imperative, we use an epidemiologic model to estimate the potential impact of existing or potential medical and public health interventions that might decrease the incidence of functional decline. For at least three major causes (stroke, hip fracture, and incontinence) of disability, the potential exists for reducing the incidence and burden of functional disability by a number of mechanisms. For example, treating just half of adults age 65-74 with currently untreated diastolic or isolated systolic hypertension would reduce the incidence of stroke by 2.77% in this age group (or 1,500 fewer cases of stroke annually). The estimates presented indicate the need (1) to better implement those interventions that are known to be efficacious, and (2) to identify and to test new interventions for conditions contributing to functional impairment in the elderly. PMID- 8418129 TI - Nursing home residents: the stories of their lives. PMID- 8418131 TI - Rotator cuff tears. PMID- 8418130 TI - Assisted suicide is not voluntary active euthanasia, but it's awfully close. PMID- 8418132 TI - Toradol and the risk of gastrointestinal complications in the elderly. PMID- 8418133 TI - DNA repair in congenic mice: possible influence of a chromosome 4 genetic region on the rate of benzo[a]pyrene-induced DNA adduct removal. AB - An attempt was made to assign mouse lifespan-associated interstrain differences in DNA repair to a specific chromosomal region using a set of congenic mice. The sensitive 32P-postlabeling assay was employed to measure the removal of benzo[a]pyrene-induced DNA adducts in liver DNA of three different chromosome 4 congenic mouse strains: B6.C-H-15c, B6.C-H-16c, and B6.C-H-26c and the two parental strains, C57B1/6 and BALB/c. The removal of the one main adduct detected, trans-(7R)-N2-[10-(7 beta,8 alpha,9 alpha-trihydroxy)-7,8,9,10- tetrahydrobenzo(a)-pyrene]-yl-deoxyguanosine (BPDE-N2-dG), in liver DNA of C57Bl/6 and BALB/c mice between one and three days after treatment, was approximately 86% and 57%, respectively. The percentage removal of BPDE-N2-dG in two of the three congenic mouse strains, B6.C-H-16c and B6.C-H-26c, resembled that found in BALB/c, whereas the third strain, B6.C-H-15c, removed about the same amount as C57B1/6, i.e., approximately 88% of BPDE-N2-dG between one and three days after treatment. The usefulness of congenic mouse strains for identifying genes putatively involved in aging and/or disease susceptibility is discussed. PMID- 8418134 TI - Dietary restriction of adult male rhesus monkeys: design, methodology, and preliminary findings from the first year of study. AB - Dietary restriction (DR) retards aging processes and extends maximum life span in rodents and in simpler animals. We initiated a study in 30 adults (8-14 years old) male rhesus monkeys to determine whether or not aging processes are retarded by adult-onset DR in a primate species and herein report results from the experiment's first year. Following a 3-6 month period when baseline data were obtained, 15 animals were assigned to a control group and given free access to a semipurified diet for 6-8 hours per day. The other 15 monkeys were fed the same diet but at 70% of their baseline intake levels predetermined individually. The animals are being evaluated semi-annually for body size and composition, physical activity, metabolic rate, glucose tolerance and insulin sensitivity, hematologic indices, immunologic function, and fingernail growth. Ocular function is assessed annually. The preliminary observations after one year are: (a) all monkeys appear to be in excellent health; (b) average body weights for controls increased by 9% while monkeys on DR did not gain weight; (c) monkeys on DR have less body fat than do control monkeys, whereas the amount of lean body mass has not been significantly influenced by DR; (d) there was a small but statistically significant reduction in physical activity for monkeys on DR relative to controls; and (e) DR has not overtly influenced the other measures. Control monkeys gradually reduced their voluntary levels of food intake during the first year of study, and food allotments to DR monkeys are being adjusted accordingly in order to reinstate the intended 30% difference between groups. These early data indicate that DR can be safely instituted in adult monkeys, but that longer term and/or more severe DR is required to determine if it is capable of influencing age-sensitive indices in long-lived primates. PMID- 8418135 TI - Diet and the suitability of the male Fischer 344 rat as a model for aging research. AB - There has been concern about the suitability of the male Fischer 344 (F344) rat as a model for aging research because of the high prevalence of a single disease, severe nephropathy, at advanced ages which confounds the interpretation of an aging study. In a publication from our laboratory, Iwasaki et al. (1988) reported that replacing the casein in our standard semisynthetic diet with soy protein markedly decreases the progression of nephropathy with advancing age in ad libitum fed male F344 rats. In the present study, it is shown that replacing the casein with lactalbumin does not decrease the occurrence of severe nephropathy in ad libitum fed rats. It is also shown that dietary restriction (DR) studies can be effectively executed in the male F344 rat when soy protein is the source of dietary protein. It is further shown that when the energy intake of the rats fed soy protein-containing diets was reduced to 60% of the ad libitum intake, almost one-third of the rats died with an absence of severe morphologic lesions, that is, lesions which contribute to the death of the rat. It is concluded that the male F344 rat is an excellent model for aging research when soy protein is the source of dietary protein; no single disease process was found to be primarily responsible for death with such a diet. PMID- 8418136 TI - The influence of an aging immune system on cancer incidence and progression. AB - The majority of cancers occur in patients over the age of 65 years, and yet the important interactions of normal aging upon the neoplastic process remain to be thoroughly explored. For example, there is no unifying and conclusive explanation for the increased risk of cancer with advancing age. The age-associated decline in immune function has previously been implicated in this regard. However, it is likely that other factors, such as the time it takes to proceed through the multistep neoplastic process, and the combined influence of age-associated increased susceptibility to carcinogen and decreased DNA repair are more important. Nevertheless, immune senescence does influence various aspects of tumor growth and spread. In this article these aspects are reviewed. PMID- 8418137 TI - A log-linear relationship between reported impairments and age: implications for the multistage hypothesis. AB - The relationship of mortality rates with age can be represented by a log-linear (Gompertz-type) plot and also, slightly less satisfactorily, by a log-log plot. Similar relationships are observed for incidence rates for a number of specific diseases. For a case where the log-log plot seems preferable, a multistage model is often invoked to explain this relationship. This study investigated the relationship between rates of various impairments and age. Both log-linear and log-log plots of data from the HALS study are compared. Because impairment rates are essentially prevalence rates, models explaining linearity in these plots are different from those used for incidence rates; however, a multistage model with independent events occurring simultaneously may apply. PMID- 8418138 TI - Aging and cancer--another perspective. PMID- 8418139 TI - Measuring functioning and health in the very old. AB - To assess the validity of brief multidimensional measures of health, we studied 155 new residents of a long-term care institution. We collected self-reported measures of various aspects of health, as well as performance-based measures of physical and cognitive function. For six similar health dimensions measured using two self-reported methods, the average correlation between paired health dimensions was 0.64 (nonpaired correlation = 0.36). When we compared self-reports and performance on three closely paired health concepts, the average correlation of paired concepts was 0.49 (nonpaired correlation = 0.22). In a factor analysis, similar health dimensions measured using different methods tended to load on the same factor. Except for manual performance, performance-based and self-reported measures of physical and role function loaded on one factor. We conclude that brief measures of health and self-reported physical functioning in very old persons have acceptable validity. PMID- 8418140 TI - Physical measurements in an elderly black population: knee height as the dominant indicator of stature. AB - This investigation examined (a) select physical measurements for age and gender differences in an elderly Black population, and (b) the relationship of stature as a dependent variable on knee height, weight, age, and gender in this population. T-test values were greater for men than for women in stature and knee height (p < .001), whereas gender differences in weight were not significant. Multiple regression was used to determine the relationship of stature to the four independent variables. When all Black subjects (n = 119) were included, three variables were significant: 3 square root knee height, gender, and 3 square root weight (R2 = .49). The equation inclusive only of the 98 Black females contained two variables: knee height2 and 3 square root weight (R2 = .29). The probability of both R2's was < .001. In decreasing order, knee height, gender, and weight were important in predicting stature; age was not significant. PMID- 8418142 TI - Agency for Health Care Policy and Research guidelines for pressure ulcer prevention: improving practice and a stimulus for research. PMID- 8418141 TI - Age, gender, medical treatment, and medication effects on smell identification. AB - Olfactory function diminishes with increasing age, which may impact on the safety and quality of life of older persons. This study examined the influence of age, gender, medical treatment, and medications on smell identification in a group of generally healthy individuals. Males (n = 221) and females (n = 166) between the ages of 19 and 95 years, from the oral physiology component of the Baltimore Longitudinal Study of Aging, were administered the University of Pennsylvania Smell Identification Test [UPSIT; (1)]. Older subjects had lower UPSIT scores, were more likely to be anosmic, and more often complained about their sense of smell and taste compared to younger subjects. Females had higher UPSIT scores compared to males. Although these age and gender effects are comparable to previous findings, performance among all subjects was superior to other studies; this is attributable, in part, to the overall healthy status of the population. Subjects being treated for medical problems and taking prescription medications had slightly lower UPSIT scores and more smell and taste complaints compared to other subjects, but these findings did not achieve statistical significance. Results suggest that age-related declines in olfaction occur as a part of the normal physiological aging process. PMID- 8418143 TI - Dermal blood flow response to constant pressure in healthy older and younger subjects. AB - This study compared the dermal blood flow response to low levels of compressive pressure in healthy older and younger subjects. Dermal blood flow was measured over the left trochanter of 19 younger (21-45 yrs) and 22 older (> or = 60 yrs) subjects using a laser-Doppler velocitometer. Interface pressure, baseline flow, and flow during 60 minutes of left-side lying on an air mattress were measured. Baseline dermal blood flow did not differ significantly between the older and younger groups. Both groups showed a significant increase in blood flow over time during the 60 minutes of compression. The change in dermal blood flow over time was not significantly different between the two groups. This study demonstrated that healthy persons, regardless of their age, exhibited an increase in dermal blood flow over time at low levels of compressive pressure. PMID- 8418144 TI - Age differences in cognitive performance in later life: relationships to self reported health and activity life style. AB - The predictive relationships among individual differences in self-reported physical health and activity life style and performance on an array of information processing and intellectual ability measures were examined. A sample of 484 men and women aged 55 to 86 years completed a battery of cognitive tasks measuring verbal processing time, working memory, vocabulary, verbal fluency, world knowledge, word recall, and text recall. Hierarchical regression was used to predict performance on these tasks from measures of self-reported physical health, alcohol and tobacco use, and level of participation in everyday activities. The results indicated: (a) individual differences in self-reported health and activity predicted performance on multiple cognitive measures; (b) self-reported health was more predictive of processing resource variables than knowledge-based abilities; (c) interaction effects indicated that participation in cognitively demanding activities was more highly related to performance on some measures for older adults than for middle-aged adults; and (d) age-related differences in performance on multiple measures were attenuated by partialing individual differences in self-reported health and activity. PMID- 8418145 TI - The impact of long-term exercise training on psychological function in older adults. AB - The effect of long-term aerobic training on psychological function was examined in 87 sedentary older adults who engaged in a year-long endurance exercise training program compared with a nonexercising control group. In addition to improved cardiovascular fitness, a positive change in self-reported morale was found for the exercise condition. Of the cognitive functions measured, a significant effect was noted for the Wechsler Memory Scale (WMS) Logical Memory subtest; however, this effect was caused by a decline in performance from pre- to posttesting in the control group. Long-term exercise training had little, if any, effect on improving cognitive function in this older adult sample. PMID- 8418147 TI - Speed and knowledge as determinants of adult age differences in verbal tasks. AB - Two studies were conducted to determine the relative importance of processing speed and knowledge as predictors of performance in simple verbal tasks within samples of young and old adults. Eight different criterion tasks were investigated, and performance on each was found to be significantly related both to speed of processing and to quantity of word knowledge. It was also discovered that although young adults were faster than old adults and that old adults were equal or superior to young adults in relevant knowledge, the same regression equations could be used to predict criterion performance in both groups. These results therefore suggest that any age-related compensation that exists in these tasks is rather weak, in the sense that speed and knowledge appear to have the same importance in young and old adults, and only the average levels of the predictors differ as a function of age. PMID- 8418146 TI - The influence of health on family contacts and family feelings in advanced old age: a longitudinal study. AB - This study provides a longitudinal analysis of the influences of health, age, gender, and socioeconomic status on family contacts and family feelings in a sample of 62 members of the Berkeley Older Generation Study. Stability in family contacts and in family feelings was observed over 14 years of advanced old age. Of the four predictor variables, health and socioeconomic status accounted for the largest proportion of observed variance. Contrary to our hypotheses, study participants in better health had greater amounts of contact with family than did those in poorer health. The former also had more feelings of closeness to family members, a finding that may reflect greater possibilities for reciprocity between elders in good health and their family members. PMID- 8418148 TI - Aging and memory for schematically vs taxonomically organized verbal materials. AB - Two experiments examined age differences in the effects of type of organization on recall of verbal materials. Young and older adults studied a list of simple verb-noun phrases that were organized either taxonomically by categories or schematically by activities, and then were tested for recall. Relative to taxonomic organization, schematic organization was found to facilitate recall in both age groups, although the effect of organization was only present for the older adults when explicit organizational cues were presented. The enhancement associated with schematic organization was attributed to the additional temporal and causal connections between items. These associations were assumed to be responsible for the facts that, relative to taxonomic organization, schematic organization was associated both with more correspondence between input and output orders and with more items being retrieved the first time an activity or category grouping was accessed at recall. PMID- 8418149 TI - Age and incidental recall for a simulated everyday memory task. AB - We compared intentional learning on an everyday memory task, associate learning of name-face pairs, with Type II incidental recall of the city of residence for each of the name-face pairs. Performance on intentional and incidental learning was significantly associated with age and performance differences begun as early as the fifth decade. Age was more strongly associated with intentional than with incidental learning, and performance on the single incidental recall trial was most similar to the first trial of the intentional learning task. PMID- 8418150 TI - Support and interpersonal stress in the social networks of married daughters caring for parents with dementia. AB - Data collected on 95 married daughters and 1,195 members of their social networks are used to investigate factors differentiating individuals who were and were not a source of social support or interpersonal stress to women caring for parents with dementia. Reports by the caregivers indicated that siblings and friends were almost equally important sources of support, but that siblings were overwhelmingly the most important source of interpersonal stress. Multivariate analyses demonstrated the importance of some dimensions of status similarity in explaining which network members were a source of support or stress. Individuals who had cared for an elderly relative were more likely to have been a source of support and less likely to have been a source of stress; individuals who were more similar in age to the caregivers were also less likely to have been a source of stress. Gender similarity was also associated with the provision of support; however, gender-similar network members were also a greater source of interpersonal stress. PMID- 8418151 TI - Stability and improvement of health after nursing home admission. AB - This prospective study investigated stability of residents' mental status, function (activities of daily living), and mood during the first and second week following nursing home admission. New residents (N = 647) in eight urban nursing homes were assessed three times. Mental status was measured using the Short Portable Mental Status Questionnaire, function was measured using Scaled Outcome Criteria, and mood was measured using the Mood Adjective Checklist. A repeated measures multivariate analysis of variance, controlling for admission from hospital and readmission to the nursing home, documented stable mental status during the first and second week of nursing home residence. Hygiene, grooming, dressing, and transferring improved, whereas feeding, ambulation, urination, and defecation were stable. Tired and depressed mood adjectives improved, and angry, cheerful, afraid, lonely, and alert mood adjectives were stable. Residents admitted from a hospital had lower mental status and function scores, and readmitted residents had lower urination scores. PMID- 8418152 TI - Physicians' ratings of health in middle and old age: a cautionary note. AB - Physicians' global assessments traditionally have been considered to be relatively objective estimates of older people's health against which self ratings of health and other self-reported measures are compared. Using data on middle-aged and older Mexican Americans from the Hispanic Health and Nutrition Examination Survey, it is suggested that the subjective component of physicians' assessments can create problems with validity, especially when a small number of physicians perform the assessments, as was the case in this study. It is recommended that researchers should not always assume that physicians' assessments represent an objective "gold standard" for validating self-reported measures of health. PMID- 8418153 TI - Replication of the multidimensionality of activities of daily living. AB - We attempted to replicate the three-dimensional factor structure of a previously proposed ADL scale and demonstrate an association between the advanced ADL dimension and cognitive function. Data used in these analyses were baseline assessments of health and functional status of hospitalized patients enrolled in a randomized controlled trial of case managers as a means of reducing health care utilization. We submitted 14 items from the OARS to a two-stage process of principal components factor analysis. Four significant dimensions emerged that were remarkably similar to the advanced, basic, and household ADL dimensions reported by Wolinsky and Johnson (1991). In this sample of hospitalized patients, however, incontinence emerged as a weak fourth dimension. Multiple regression of SPMSQ mental status examination scores on these ADL dimensions demonstrates the association between cognitive function and the advanced ADL dimension. These data confirm that the underlying structure of ADLs consists of at least three separate dimensions, one of which is aligned with cognitive capacity. PMID- 8418154 TI - On black burden and becoming nouveau poor. PMID- 8418155 TI - Gender differences in parent care: demographic factors and same-gender preferences. AB - This study tested the hypothesis that, when aging parents are assisted in the tasks of daily living by their adult children, the gender of the child providing care depends in part upon the gender of the parent requiring care. Data on 4,371 infirm elders and their 13,172 adult children from the National Long-Term Care Survey were analyzed by means of logistic regression. Separate analyses show that adult children are more likely to provide care to a parent of the same gender, and infirm elders are more likely to receive care from a child of the same gender. Because the substantial majority of elderly parents requiring care from children are mothers, this tendency toward gender consistency in the caregiving relationship partially accounts for the fact that daughters are more likely than sons to be involved in parent care. PMID- 8418156 TI - Role of egg consumption in sporadic Salmonella enteritidis and Salmonella typhimurium infections in Minnesota. AB - The incidence of Salmonella enteritidis infections has increased in the eastern United States, and consumption of undercooked eggs has been associated with outbreaks of S. enteritidis. In Minnesota, the incidence of S. enteritidis infections doubled from 1980 to 1990; however, no egg-associated outbreaks were identified. A case-control study was conducted to examine potential exposures for S. enteritidis and Salmonella typhimurium infections in Minnesota adults. Sporadic cases of S. enteritidis (odds ratio [OR], 5.2; 95% confidence interval [CI], 1.9-14.2; P = .003) and S. typhimurium infection (OR, 2.4; CI, 1.1-5.5; P = .03) were more likely to have consumed undercooked eggs or egg-containing foods during the 3 days before onset of illness compared to a similar reference period for controls. In addition, the extent to which eggs were cooked was directly associated with illness (chi 2 test for trend, P < .001). These findings demonstrate that eggs are important vehicles for S. enteritidis and S. typhimurium, even in the absence of recognized outbreaks. PMID- 8418157 TI - The efficacy of combination immunotherapy in experimental Pseudomonas sepsis. AB - Immunotherapy has been shown to be an effective adjuvant in the management of septic shock. A neutropenic rat model of septic shock induced by infection with Pseudomonas aeruginosa 12.4.4 (Fisher immunotype 6) was used to determine the relative efficacy of single, double, and triple combination immunotherapy. A Pseudomonas O serotype-specific, opsonophagocytic monoclonal antibody (MAb), polyclonal J5 antiserum, and a MAb directed against tumor necrosis factor-alpha (TNF) were studied as single therapy and in combination. The combination of all three immunotherapeutic agents resulted in a 77% survival rate (33/43 animals). This level of protection was superior to that achieved with any combination of two antibody treatments (50%-60% survival; P = .029) or single antibody therapy (25%-43% survival; P < .001) or compared with a control group (0/25 survivors; P < .0001). Immunotherapy directed against multiple steps of the septic process is more active than single or double antibody regimens and may offer an improved approach to the adjunctive treatment of septic shock. PMID- 8418158 TI - Protective activity of anti-exotoxin A monoclonal antibody against mice infected with toxin-producing Pseudomonas aeruginosa. AB - The neutralizing and protective effect of murine monoclonal antibody (MAb) 3C7 (IgG1) against Pseudomonas aeruginosa exotoxin A was examined in an experimental mouse model of infection with exotoxin A-producing strains. Treatment with MAb 3C7 blocked the reduction of functional elongation factor 2 (EF-2) in the liver of mice but could not clear the bacteria. Administration of gentamicin caused bacteria to be cleared but did not block reduction of hepatic EF-2 level. Treatment with either MAb 3C7 or gentamicin individually did not prolong time to death; however, the combined therapy with both MAb 3C7 and gentamicin cleared bacteria and blocked the reduction of hepatic EF-2 level, resulting in a significant increase in the survival rate of mice. These results suggest that anti-exotoxin A MAbs show effectiveness against pseudomonal infection caused by exotoxin A-producing strains. PMID- 8418159 TI - Efficacy of anti-endotoxin monoclonal antibody E5 alone or in combination with ciprofloxacin in neutropenic rats with Pseudomonas sepsis. AB - Pathogen-free rats were rendered neutropenic, given oral feedings of Pseudomonas aeruginosa 12.4.4, then monitored for fever. At the onset of fever, rats were given intravenous treatment with either anti-endotoxin monoclonal antibody (MAb) E5 or control MAb B55. Survival was significantly greater in E5- than in B55 treated animals (P < .01). Serum levels of both lipopolysaccharide and tumor necrosis factor-alpha were significantly reduced in E5- versus B55-treated rats 24 h after treatment (P < .01 and < .05, respectively). Rats were also treated with E5 or B55 in combination with a suboptimal dose of ciprofloxacin at fever onset and again 24 h later. Survival was significantly greater in ciprofloxacin treated animals given E5 than in animals given B55 (P < .005). Posttreatment endotoxin levels were decreased in animals receiving E5 in combination with ciprofloxacin (P < .001) compared with B55-treated animals. These results indicate that therapy with anti-endotoxin MAb E5 alone or in combination with antimicrobial therapy improves survival in this bacteremic infection model of Pseudomonas sepsis. PMID- 8418160 TI - Nontoxic forms of lipopolysaccharides, poorly cleared in vivo, are present in variable amounts in phenol-extracted endotoxins. AB - A radiolabeled lipopolysaccharide (LPS) from Salmonella choleraesuis was as toxic for galactosamine-treated mice as the unlabeled preparation. After intravenous injection, a large proportion (37%) of the labeled material remained in the circulation. This circulating form of LPS was not toxic, even when reextracted with phenol, and remained in the circulation when reinjected into mice. Furthermore, toxic and nontoxic constituents were directly isolated in vitro by hydrophobic gel chromatography of the radiolabeled LPS preparation. The nontoxic form, of lower molecular weight according to its electrophoretic migration, did not react with an anti-lipid A monoclonal antibody but reacted with polymyxin B and was as active on pre-B and B cells as the unfractionated LPS preparation. The results of this study suggest that some LPS constituents of phenol-extracted endotoxin preparations are not toxic for galactosamine-sensitized mice and are not trapped by appropriate LPS-binding sites in the liver as efficiently as are the toxic forms. PMID- 8418161 TI - Evaluation of molecular methodologies and rabbit infectivity testing for the diagnosis of congenital syphilis and neonatal central nervous system invasion by Treponema pallidum. AB - IgM immunoblotting and polymerase chain reaction (PCR) were evaluated for use in diagnosing congenital syphilis, and the prevalence of central nervous system (CNS) invasion by Treponema pallidum during congenital infection was examined. The results of rabbit infectivity testing (RIT) on serum and cerebrospinal fluid (CSF) of 19 infants born to mothers with untreated early syphilis were compared with results of PCR and IgM immunoblotting. Seven infants had clinical evidence of congenital syphilis supported by positive serum IgM immunoblot (7/7), PCR (6/7), and RIT (3/3). Six symptomatic infants (86%) had T. pallidum isolated from CSF by RIT; 5 of 6 RIT-positive CSF samples were positive by PCR, and 2 also were reactive by IgM immunoblot. In 12 asymptomatic infants, 5 (42%) had a reactive serum IgM immunoblot and in 4 of these IgM reactivity was the only evidence of congenital infection. CNS invasion by T. pallidum was uncommon among asymptomatic infants; only 1 (8%) was positive by CSF RIT. The excellent agreement between RIT and PCR further substantiates the use of PCR as a surrogate for RIT. Our data indicate that the diagnosis of asymptomatically infected neonates will require a comprehensive approach using assays for both specific neonatal IgM and T. pallidum DNA in serum and CSF. PMID- 8418162 TI - Characterization of the borreliacidal antibody response to Borrelia burgdorferi in humans: a serodiagnostic test. AB - An in vitro assay was used to characterize the borreliacidal activity of sera from Lyme disease patients. The mean percentage of killing was 23% with sera from patients with a single erythema migrans lesion, 42% from patients with multiple lesions, 58% from patients with Lyme arthritis of short duration, and 83% from patients with Lyme arthritis of long duration. Borreliacidal activity was abrogated when Lyme disease serum was treated with anti-human IgM or IgG1. In addition, human sera from Lyme arthritis patients containing borreliacidal antibody prevented the induction of Lyme arthritis in irradiated hamsters challenged with the Lyme spirochete. Removal of outer surface protein A antibodies from late Lyme disease sera caused reductions in the borreliacidal antibody titer. The results demonstrate an important role for borreliacidal antibody against infection with B. burgdorferi in humans and confirm that detection of borreliacidal antibody in human sera can be a specific serodiagnostic test for Lyme disease. PMID- 8418163 TI - In vitro inhibition of Borrelia burgdorferi growth by antibodies. AB - Function-oriented immunoassays, such as complement fixation and neutralization, are not commonly used in the study of the Lyme disease agent, Borrelia burgdorferi. To determine whether such assays provide information additional to matrix-based methods, such as ELISA, polyclonal antisera and monoclonal antibodies were examined for their abilities to agglutinate viable borreliae and inhibit their in vitro growth in microtiter plates. Different strains of B. burgdorferi in both high and low passage were examined, and the related spirochete Borrelia hermsii and antibodies to it served as controls. Agglutination and complement-independent inhibition of growth with polyclonal sera from rats, mice, and humans and with monoclonal antibodies to outer membrane proteins OspA and OspB was demonstrated. Growth inhibition was obtained with Fab fragments as well as with whole IgG molecules. In comparison with an ELISA using whole cells, the growth inhibition and agglutination assays were generally more specific. PMID- 8418164 TI - Incidence of and significant risk factors for aminoglycoside-associated nephrotoxicity in patients dosed by using individualized pharmacokinetic monitoring. AB - Incidence of and risk factors for aminoglycoside-associated nephrotoxicity (AAN) were evaluated in 1489 patients prospectively monitored with individualized pharmacokinetic monitoring (IPM). Incidence of AAN was 7.9% with individual (univariate) risk factors including advanced age, decreased albumin, poor nutritional status, pneumonia, concurrent furosemide, amphotericin B, vancomycin, cephalosporin, or piperacillin, intensive care unit treatment, leukemia, rapidly fatal illness, liver or renal disease, reduced aminoglycoside clearance, elevated initial steady-state trough concentration (Cminss), volume of distribution or half-life, duration of therapy, total dose, fever, male gender, shock, pleural effusion, and ascites. Multiple logistic regression revealed that Cminss, concurrent clindamycin, vancomycin, piperacillin, or cephalosporin, ascites, advanced age, male gender, decreased albumin, duration of therapy, and leukemia were significant independent risk factors for AAN. Positive predictive value of the model was 30.8%; negative predictive value was 91.7%. No identifiable risk factor alone or in combination was of sufficient sensitivity to reliably predict AAN, but use of IPM may lower the incidence of AAN. PMID- 8418165 TI - Regulation of cryptococcal capsular polysaccharide by iron. AB - Iron is tightly controlled in mammalian tissues and regulates virulence factors in various pathogenic organisms. The influence of Fe availability upon production of cryptococcal capsular polysaccharide was studied. Polysaccharide, measured as cell-bound glucuronyl residues, increased more than threefold as available Fe in the culture medium was varied from repletion to tight sequestration and depletion in five incremental steps. Since physiologic CO2 concentration may serve as stimulus for cryptococcal polysaccharide synthesis, the combined effect of Fe availability and CO2 on encapsulation was studied. Addition of dissolved, loosely chelated Fe moderated the effect of CO2. Tight chelation of dissolved Fe potentiated the CO2 effect. Tissue from infected mice showed heavily encapsulated organisms, consistent with results with physiologic CO2 concentration and Fe deprivation. In conclusion, cryptococcal polysaccharide synthesis is increased by limitation of ferric iron availability to the cell and by dissolved CO2, and the two effects are additive. PMID- 8418166 TI - Immunogenicity and safety of respiratory syncytial virus subunit vaccine in seropositive children 18-36 months old. AB - Twenty-six children (aged 18-36 months) previously hospitalized for respiratory syncytial virus (RSV) infection were randomized to receive 50 micrograms of an RSV subunit vaccine composed primarily of F glycoprotein or saline placebo by intramuscular injection. Serum was obtained at entry and at 1 and 6 months after vaccination for detection of antibody to F glycoprotein and G glycoprotein of subtypes A (Ga) or B (Gb) and of neutralizing antibody (nAb). At 1 month, by comparing the baseline values, vaccinees had statistically significant increases in geometric mean antibody titer (GMT) of more than fourfold to F (P = .0001), Ga (P = .0001), Gb (P = .003), and nAb (P = .009). No differences in GMT were observed between F protein vaccine and placebo recipients at entry, nor between placebo recipients at entry and 1 month. RSV infections were identified in 7 placebo recipients (4 by both viral identification and seroconversion, 3 by seroconversion alone). No vaccine recipient had RSV infection documented in the 6 months after vaccination (P = .003). There were no significant vaccine-related side effects, and no evidence of enhanced respiratory illnesses was observed. The subunit F protein vaccine appears safe and immunogenic and may prevent infection in healthy children primed by prior RSV infection. PMID- 8418167 TI - Effect of a hepatitis B vaccination program on the prevalence of hepatitis B virus infection. AB - In April 1991, surveys for serologic evidence of hepatitis B virus (HBV) infection were conducted among 3- to 4-year-old children born after a hepatitis B immunization program of newborns began and among 6- to 11-year-old children targeted for early childhood vaccination in American Samoa. Compared with 3- to 4 year-olds tested in 1991, children tested at baseline in 1985 were more likely to have been infected with HBV (5/40 vs. 2/93; prevalence ratio [PR] = 5.8, 95% confidence limits [CL] = 1.2, 28.7) and to have chronic infection with HBV (3/40 vs. 0/95; PR = undefined, lower CL = 1.2). Compared with 6- to 11-year-olds tested in 1991, children in 1985 were more likely to have been infected with HBV (32/121 vs. 53/386; PR = 1.9, CL = 1.3, 2.8) and to have chronic infection with HBV (8/121 vs. 7/386; PR = 3.6, CL = 1.3, 9.8). The incorporation of hepatitis B vaccine into routine childhood vaccination schedules can prevent acute and chronic HBV infection in areas of high endemicity. PMID- 8418168 TI - Diagnosis of rabies by polymerase chain reaction with nested primers. AB - A simple, sensitive, and specific polymerase chain reaction (PCR) protocol for detection of rabies virus is described. The process consists of sample preparation, reverse transcription, two-step DNA amplification, and detection of the amplified product. RNA was extracted from animal and human brain by phenol chloroform using guanidinium thiocyanate. Viral RNA was then amplified in a two step PCR that used two sets of nested primers designed to amplify rabies nucleocapsid (N) sequence. Rabies nucleocapsid sequence was amplified from all brain samples from 95 dogs and 3 humans with rabies confirmed by fluorescent antibody (FAT) and mouse inoculation tests (MIT). Rabies-negative brain samples (110 dogs, 2 humans) were PCR-negative. The process requires < 24 h. Detection of viral RNA was still possible in brain material that was left at room temperature for 72 h. As little as 8 pg of rabies virus RNA could be detected. This technique could have practical applications as a confirmatory test to FAT at busy rabies diagnostic centers. PMID- 8418169 TI - Infectious decay of human immunodeficiency virus type 1 in plasma. AB - AIDS is caused by the human immunodeficiency virus type 1 (HIV-1). Recent methods have been developed to estimate infectious titer in various bodily fluids, including blood. However, lack of information about HIV-1 stability in blood has restricted the use of these techniques to fresh samples in immediately accessible virology laboratories. In studies of infectious virus decay, it was found that at room temperature, complete decay of infectious HIV-1 in plasma can require > 7 days. Furthermore, the stability of HIV-1 was enhanced by storage at 4 degrees C, suggesting that fresh plasma could be sent on ice to core laboratories for viral quantitation. These studies also emphasize the need for thorough decontamination of all potentially infectious material. PMID- 8418170 TI - Quantification of human immunodeficiency virus type 1 tat mRNA as a marker for assessing the efficacy of antiretroviral therapy. AB - To evaluate the use of human immunodeficiency virus type 1 (HIV-1) tat mRNA quantification as a marker for antiretroviral therapy, 10 zidovudine-naive, p24 antibody-positive subjects (Centers for Disease Control classes III and IV) were studied at the start of zidovudine treatment. HIV-1 proviral DNA content and tat mRNA levels were monitored for 20 weeks from the initiation of therapy. Levels of tat mRNA were quantified using an engineered tat cRNA internal control under conditions of linear amplification. Proviral DNA levels increased in 2 patients and decreased in 8 during 20 weeks of therapy. In each case, tat mRNA levels exhibited similar but much more pronounced changes. Results indicate that quantitative measurement of tat mRNA levels is extremely useful for monitoring antiretroviral therapy. PMID- 8418171 TI - Intestinal microsporidiosis in human immunodeficiency virus-infected patients with chronic unexplained diarrhea: prevalence and clinical and biologic features. AB - Eighteen patients infected with human immunodeficiency virus and with chronic unexplained diarrhea were prospectively studied to investigate the prevalence and clinical and biologic features of intestinal microsporidiosis. All patients underwent extensive evaluation for bacterial, viral, and parasitic pathogens. Enterocytozoon bieneusi was found in 9 patients (50%; 95% confidence interval, 27 73) in stools and duodenal and jejunal biopsies. In 8 patients, it was the sole pathogen found. Other pathogens were also isolated from the intestinal tracts of 4 patients, but diarrhea remained unexplained in 6. Patients with intestinal microsporidiosis had significantly lower mean Karnofsky scores (69.4 vs. 85.5, P = .009), CD4 cell counts (18.6 vs. 209.8/microL, P = .02), and D-xylose absorption tests (0.13 vs. 0.36 g/L, P < .001) than did patients without intestinal microsporidiosis. Intestinal microsporidiosis appears to be a frequent cause of unexplained chronic diarrhea in patients with AIDS and is associated with diminished D-xylose absorption. PMID- 8418173 TI - Pseudomonas aeruginosa colonization of the gastrointestinal tract in patients with cystic fibrosis. AB - Respiratory and fecal specimens from 111 patients with cystic fibrosis (CF) were cultured on 394 occasions to determine which site Pseudomonas aeruginosa colonizes first. By an enrichment and selection culture technique, P. aeruginosa was recovered from fecal cultures in 21 (42%) of 50 respiratory tract-positive patients but from only 3 (6.3%) of 48 respiratory tract-negative patients and from 1 (2.2%) of 45 control subjects. In 4 (22%) of the 18 patients who became culture-positive during the study, P. aeruginosa was recovered from a fecal specimen before respiratory colonization. Only 1 of these 4 became persistently colonized with P. aeruginosa in the respiratory tract; however, the serotype of the respiratory isolate was different from that of the fecal isolate. P. aeruginosa is recovered rarely from the feces of patients with CF who are culture negative in the respiratory tract. In patients with CF, primary colonization with P. aeruginosa probably occurs in the respiratory tract. PMID- 8418172 TI - Variable region-identical monoclonal antibodies of different IgG subclass directed to Pseudomonas aeruginosa lipopolysaccharide O-specific side chain function differently. AB - Antibodies directed to polysaccharide (PS) antigens of bacteria are crucial to host immunity to infection. The isotypes of antibodies made to PS, however, are restricted primarily to IgM and IgG1 and IgG2 in man and to IgM and IgG3 in mice. Using sequential sublining and sib selection, an IgG1 murine monoclonal antibody that has variable regions identical to those of a parent IgG3 monoclonal antibody directed to the high-molecular-weight component of the O-specific side chain of Pseudomonas aeruginosa immunotype 1 lipopolysaccharide was derived. These antibodies differed markedly in their antigen binding and effector functions. IgG3 was superior in binding to multivalent PS both in purified and whole bacterial form, fixation of the third component of complement to the bacterial surface, and opsonization of P. aeruginosa for uptake by both murine and human phagocytes. These data suggest that the IgG subclass of these murine anti-LPS antibodies is an important determinant of both avidity for multivalent antigen and biologic function. PMID- 8418174 TI - Chemotactic activity of urethral secretions in men with urethritis and the effect of treatment. AB - The chemotactic activity of urethral exudate and the effect of treatment were assessed in 67 patients with nongonococcal urethritis (NGU) or gonorrhea. All samples demonstrated chemotactic activity related to the number of neutrophils seen on Gram's staining of the urethral smear. Chemotactic activity decreased after treatment in patients with Chlamydia-positive NGU (mean +/- SE, 21.1 +/- 2.7 cells/high-power field [hpf] before treatment; 8.7 +/- 2.2 after treatment; P < .004) and gonorrhea (25.6 +/- 5.2 cells/hpf before treatment; 1.6 +/- 0.7 after treatment; P < .015). Chemotactic activity increased again in Chlamydia-positive NGU patients to 21.0 +/- 3.3 cells/hpf (P < .05) 2-3 weeks after cessation of therapy in the absence of demonstrable infection or further intercourse. There was no significant decrease in chemotactic activity after therapy in the Chlamydia-negative NGU or persistent urethritis groups. These data suggest a previously unrecognized persistent chemotactic stimulus in urethral exudate from patients with urethritis. PMID- 8418175 TI - Antibiotic-induced bacterial killing stimulates tumor necrosis factor-alpha release in whole blood. AB - Rapid lysis of gram-negative bacteria is associated with considerable release of free endotoxin. Production of tumor necrosis factor (TNF) from adult whole blood ex vivo in response to bacterial products generated during antibiotic killing of Haemophilus influenzae type b (Hib) was investigated. Heparinized whole blood released TNF in a dose-dependent fashion in response to purified lipooligosaccharide of Hib. Bacteria (10(4)-10(7) cfu/mL) were placed into a Transwell filter insert (0.1 microns) and incubated with whole blood in the presence of various antibiotics. Exposure to ceftriaxone resulted in significantly greater release of TNF during killing of Hib than did exposure to imipenem, despite similar degrees of bacterial killing at 6 h. Polymyxin B inhibited the ceftriaxone-induced TNF release by 97%-99%, indicating that free endotoxin was the predominant stimulus for the increase in TNF release in this system. These observations suggest that ceftriaxone-induced killing of Hib results in bacterial cell wall products that are more proinflammatory than those produced by imipenem. PMID- 8418176 TI - Interleukin-1 pretreatment protects against endotoxin-induced hypotension in rabbits: association with decreased tumor necrosis factor levels. AB - Rabbits were infused with either low-dose recombinant human interleukin (IL)-1 beta or vehicle 16 h prior to receiving lipopolysaccharide (LPS). In controls pretreated with vehicle, mean arterial pressure decreased to 68% of the baseline value by 90 min after LPS infusion and remained low for the duration of the study period. In IL-1-pretreated animals, a similar decrease in mean arterial pressure to 69% was observed at 60 min but returned to 90% of baseline after 180 min (P < .05). IL-1-pretreated animals showed no significant rise in serum tumor necrosis factor after LPS infusion compared with vehicle-treated animals (P < .05). Rabbits were pretreated as above with IL-1 and, 40 h later, plasma was transfused into a second group of rabbits just prior to LPS infusion. An infusion of post-IL 1 plasma did not reduce LPS-induced hypotension. Thus, pretreatment with IL-1 reduces the hypotension in response to LPS by a mechanism that is not plasma mediated but that is associated with reduced serum tumor necrosis factor levels. PMID- 8418177 TI - Foodborne giardiasis in a corporate office setting. AB - Giardiasis is the most commonly reported intestinal protozoal infection worldwide, but its relatively long incubation period and often insidious onset make detection of common-source outbreaks difficult. Few well-documented foodborne outbreaks of giardiasis have been reported. In November 1990, such an outbreak among insurance company employees resulted in 18 laboratory-confirmed and 9 suspected cases of giardiasis. A case-control study of 26 ill and 162 well employees implicated raw sliced vegetables served in the employee cafeteria and prepared by a food handler infected with Giardia lamblia as the probable vehicle (odds ratio, 5.1; 95% confidence interval, 1.4-22.7). This outbreak illustrates the potential for transmission of Giardia organisms to occur in commercial establishments through a frequently served food item. PMID- 8418178 TI - Varicella in pregnancy, the fetus, and the newborn: problems in management. PMID- 8418179 TI - Infectious Diseases Society of America guidelines for ethical conduct by members and fellows. PMID- 8418180 TI - Effect of didanosine on human immunodeficiency virus viremia and antigenemia in patients with advanced disease: correlation with clinical response. AB - To determine if suppression of human immunodeficiency virus (HIV) replication during antiretroviral therapy correlates with clinical outcome, serial quantitative serum cultures and HIV p24 antigen measurements were made in patients with advanced HIV disease treated with didanosine. Twenty-one (78%) of 27 had viremia detected, and in 14 (67%) viral titer decreased by fivefold or more. Compared with those with no decrease, patients who had a decrease in titer were more likely to achieve a > or = 5% increase in body weight (8/12 vs. 0/7, P = .013) and had a significantly greater mean increase in body weight during treatment months 1-5. Occurrence of new AIDS-defining illnesses and survival were not significantly different between groups. Changes in p24 antigenemia did not correlate with any parameter of clinical outcome examined. Changing serum HIV titer is a marker of the virologic effect of didanosine therapy that correlates with the early clinical benefit as reflected by weight gain. However, correlation of this marker with long-term clinical benefit is uncertain. PMID- 8418181 TI - High socioeconomic status is a risk factor for human immunodeficiency virus type 1 (HIV-1) infection but not for sexually transmitted diseases in women in Malawi: implications for HIV-1 control. AB - A total of 6600 pregnant urban women attending an antenatal clinic of a major hospital in Malawi were evaluated for risk factors for human immunodeficiency virus type 1 (HIV-1) infection. HIV-1 seroprevalence was 23% and significantly (P < .001) associated with markers of heterosexual activity (history of sexually transmitted diseases [STDs], more than one sexual partner, reports of husbands with other partners, and all prevalent STDs except chlamydial infection) and higher socioeconomic status (as measured by husband's education). History of injections, scarification, and transfusions were not associated with HIV-1 infection. In two logistic regression models, higher socioeconomic status was a risk factor for HIV-1 infection (odds ratio [OR] = 2.69, P < .001) but not for STDs (OR = 0.94, P = .30). The opposite associations between HIV-1 and socioeconomic status and STDs and socioeconomic status suggest that HIV prevention strategies, in addition to STD diagnosis and treatment, should include interventions to reduce high-risk sexual activity and promote condom use. PMID- 8418182 TI - Influence of tuberculosis on human immunodeficiency virus (HIV-1): enhanced cytokine expression and elevated beta 2-microglobulin in HIV-1-associated tuberculosis. AB - Tuberculosis results in activation of T cells and macrophages that may harbor latent human immunodeficiency virus (HIV-1). Although such activation is beneficial to the host in terms of mycobacterial disease, it may be deleterious in terms of HIV-1. In Ugandan HIV-1-seropositive patients with pulmonary tuberculosis, antigen-induced blastogenesis and production of tumor necrosis factor-alpha (a cytokine that induces expression of HIV-1 in latently infected cells) were 3-10 times greater than in controls. The mean serum beta 2 microglobulin level was 5.22 mg/L in recently diagnosed patients, significantly greater than levels in HIV-negative patients with tuberculosis or asymptomatic HIV-1-seropositive subjects. beta 2-microglobulin was significantly lower in subjects who had completed at least 2 months of antituberculous therapy. These observations suggest that HIV-1-associated tuberculosis is accompanied by immune activation that may result in increased HIV expression and accelerated progression to AIDS. PMID- 8418184 TI - Comparison of risk factors for hepatitis C and hepatitis B virus infection in homosexual men. AB - Serum samples from 735 homosexual or bisexual men were tested for antibodies to hepatitis C virus (HCV) and serologic markers of hepatitis B virus (HBV), and risk factors for each infection were compared. Thirty-four (4.6%) were confirmed HCV-positive compared with 81% positive for one or more HBV serologic marker(s). History of intravenous drug use (IVDU) and blood transfusion were significantly associated with HCV positivity (odds ratio [OR] = 14.3 and 4.4, respectively), but neither was significantly associated with HBV positivity. Sexual behavior was significantly associated with infection with both viruses. When IVDU and blood transfusion were controlled for, HCV infection was marginally associated with > 50 sex partners/year (OR = 2.1), > 25 oral receptive partners (OR = 2.4), and > 25 anal receptive partners (OR = 1.9). HBV infection was more strongly associated with the same variables. HCV infection is uncommon in homosexual men and IVDU is the primary route of transmission, but sexual transmission also occurs, albeit infrequently. PMID- 8418185 TI - Oral immunization with influenza virus in biodegradable microspheres. AB - Polymeric microspheres were evaluated as an oral antigen delivery system for immunization with influenza virus. The immune responses obtained were compared after either oral or systemic immunization of BALB/c mice using purified, formalin-inactivated influenza virus type A/H3N2, either encapsulated in biodegradable and biocompatible microspheres or free in solution. The immunogenicity of formalin-treated influenza vaccine was preserved during the microencapsulation process, and the microencapsulated antigen induced protective immune responses after systemic immunization that were equal to or higher than those induced by conventional vaccine. When administered orally to primed animals, microencapsulated antigen induced levels of anti-influenza antibodies in saliva that were higher than and in serum that were comparable to those obtained by systemic immunization. Furthermore, oral booster immunization provided virtually complete protection of animals challenged with live virus. PMID- 8418183 TI - Heterosexual transmission of human T cell leukemia/lymphoma virus type I among married couples in southwestern Japan: an initial report from the Miyazaki Cohort Study. AB - To identify factors that may modify the heterosexual transmission of human T cell leukemia/lymphoma virus type I (HTLV-I), 534 married couples enrolled in the Miyazaki Cohort Study between November 1984 and April 1989 were studied: 95 husband HTLV-I-seropositive (H+)/wife seropositive (W+), 33 H+/W-, 64 H-/W+, and 342 H-/W-. After 5 years of follow-up, seven seroconversions occurred and clustered significantly among serodiscordant pairs (relative risk [RR] = 41.2); the rate of transmission was 3.9 times higher if the carrier spouse was male (P = .19). Among H+/W- couples, husband's age > or = 60 years strongly predicted seroconversion in the wives (RR = 11.5). All 4 carrier husbands whose wives seroconverted had HTLV-I titers > or = 1:1024 (P = .04) and were anti-tax antibody positive (P = .06). In cross-sectional analysis, total parity also was independently associated with wife's serostatus but only length of marriage with husband's. Overall, sexual transmission of HTLV-I was primarily from older infected husbands to their wives, with husbands' viral status being an important factor. PMID- 8418186 TI - Comparison of the opsonic activity of human surfactant protein A for Staphylococcus aureus and Streptococcus pneumoniae with rabbit and human macrophages. AB - Surfactant protein A (SP-A) is a glycosylated apoprotein that may facilitate bacterial phagocytosis and contribute to early bacterial clearance in the lung. The effect of SP-A on attachment (or ingestion) of Staphylococcus aureus and type 25 pneumococci to rabbit alveolar macrophages and human monocyte-derived macrophages was studied. SP-A bound to S. aureus and type 25 pneumococci in a calcium-dependent manner. Bacteria-associated SP-A significantly increased attachment of S. aureus, but not pneumococci, to macrophages. Increased association of SP-A-coated S. aureus with macrophages appeared to consist mainly of attachment without ingestion, as determined by bactericidal tests and release of tritiated bacterial digestion products from macrophages. Preincubation of macrophages with SP-A did not increase attachment or ingestion of S. aureus or type 25 pneumococci, with or without the addition of immune opsonins. SP-A acts as a ligand to facilitate attachment of S. aureus to macrophages but has no effect on S. pneumoniae. PMID- 8418187 TI - Learning, school performance, and children with asthma: how much at risk? AB - Children with asthma may be at risk for decreased school functioning due to acute exacerbations, increased absenteeism, iatrogenic effects of their asthma medication, and the stress associated with a chronic illness. The purpose of this article is to critically review extant research pertaining to the school functioning of children with asthma, including studies of school attendance, school performance, the effects of asthma medications on learning and behavior, and the role of psychological variables in the development of functional impairments. There is not sufficient evidence to suggest that children with asthma are at significantly higher risk for poor school performance than children without asthma. Factors that may contribute to poor school performance among children with asthma include iatrogenic effects of oral steroids, poor medical management of the disease, and psychological problems. Recommendations for improving the school functioning of children with asthma are discussed. PMID- 8418188 TI - Neurocognitive aspects of pediatric sickle cell disease. AB - Studies pertaining to the neurocognitive functioning and learning of children in whom sickle cell disease is diagnosed are reviewed, and findings suggest diffuse neurocognitive deficits, with much variability across subjects. A hypothesis is presented about the cumulative nature of such deficits in children who have not sustained cerebral vascular accidents. Important methodological shortcomings in the literature are identified and recommendations are made for future neurocognitive research with children in whom sickle cell disease has been diagnosed. Studies pertaining to the psychosocial development of these children are also reviewed, and it is concluded that behavioral problems, low self-esteem, and disturbances of body image are frequently characteristic of these children. Recommendations are made including early special education and psychosocial intervention programs for children with sickle cell disease. PMID- 8418189 TI - Neuropsychological, academic, and adaptive functioning in children who survive in hospital cardiac arrest and resuscitation. AB - Children suffering cardiac arrest (CA) are not uncommon in certain pediatric populations. Due to the increasing survival rates of child CA patients, there is a growing interest in, and concern for, their long-term intellectual, academic, emotional, and adaptive functioning. This article describes the possible neurologic sequelae of CA in children and presents standardized assessment results on 25 children, 2 to 15 years of age, who survived a CA while in the hospital. A majority of these children exhibited low-average to deficient levels of performance on neuropsychologic, achievement, and adaptive behavior measures. Duration of cardiac arrest and a medical risk score were significantly correlated with decreased functioning in child CA patients. Children who suffer a cardiac arrest are at high risk for academic struggles, and many may need special education services. PMID- 8418190 TI - The write-say method for improving spelling accuracy in children with learning disabilities. AB - We evaluated a teaching strategy designed to improve spelling accuracy in children with learning disabilities. The write-say method provides immediate feedback to dual sensory modalities (i.e., visual and auditory) following the administration of a daily spelling test. Four males and three females (mean age = 11.61 years) with learning disabilities were introduced to the write-say procedure within a multiple-baseline design. Compared to control conditions (i.e., studying words on one's own), experimental procedures significantly enhanced subjects' spelling accuracy in a brief period of time. The write-say procedure was also advocated as a cost-effective method of spelling instruction for small classroom settings. PMID- 8418191 TI - Instructional practices in mainstreamed secondary classrooms. AB - This study examined the skills of secondary students that teachers considered necessary for success in mainstreamed regular education classrooms and intervention strategies teachers would be willing to use. Teachers (N = 89) from three high schools rated the importance of 14 student skills and the reasonability of 35 intervention strategies. Student skills deemed important included following directions, coming to class prepared with materials, using class time wisely, and making up assignments and tests. Factor analysis of intervention strategy results revealed seven factors: using supplemental resources; simplifying instruction; providing support and extra instructional cues; enhancing classroom behavior management; facilitating grade improvement; modifying learning environment; and teaching study skills and providing a positive, cooperative learning environment. Secondary teachers appear to prefer intervention strategies that they can implement in their own classroom, that can apply to all students, and that require little extra time. PMID- 8418192 TI - A neuropsychological approach to the Bannatyne recategorization of the Wechsler Intelligence Scales in adults with learning disabilities. AB - An investigation was conducted of the diagnostic ability of the Bannatyne recategorization of the Wechsler Adult Intelligence Scale-Revised (WAIS-R) into spatial, verbal-conceptual, and sequential components for adults with learning disabilities. A comparison among neuropsychological, intelligence, and achievement test data was made to evaluate the applicability of this recategorization. The Luria-Nebraska Neuropsychological Battery (LNNB), the WAIS R, and the Wide Range Achievement Test-Revised (WRAT-R) were administered to 103 adult subjects with learning disabilities. The LNNB scales were cluster analyzed, and cluster membership was cross-tabulated against WAIS-R, WRAT-R, and demographic data. The major findings were that (a) the mean scores fit the Bannatyne pattern in two of the four clusters; and (b) the Bannatyne pattern was found in only about 20% of the total sample. It was concluded that identification of the Bannatyne pattern is of diagnostic utility, but its absence is inconclusive. PMID- 8418193 TI - Psychoeducational characteristics of children and adolescents with insulin dependent diabetes mellitus. AB - In recent years, researchers have observed selective neuropsychological impairment associated with insulin-dependent diabetes mellitus in childhood. At increased risk are children who develop diabetes before the age of 5, who experience severe hypo- and hyperglycemia, or who have frequent episodes of mild to moderate hypoglycemia. This article explores the existing literature to establish frequency and consistency of general and specific neurocognitive deficits in this pediatric patient population, as well as the impact of these deficits on school achievement and learning disabilities. Studies are integrated to identify contributing diabetes and nondiabetes factors. This is followed by a reanalysis of the data from two studies of diabetic children to determine the learning disability characteristics of this population and the factors contributing to dysfunctional school performance. Findings are discussed in terms of the impact of different factors reflecting adequacy of diabetes control on specific psychoeducational abilities. PMID- 8418194 TI - The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. AB - Cytolytic T lymphocytes (CTL), natural killer cells, and lymphokine-activated killer (LAK) cells are cytolytic cells known to release the cytolytic protein perforin and a family of proteases, named granzymes, from cytoplasmic stores upon interaction with target cells. We now report the purification of an additional major 60-kD granule-associated protein (grp 60) from human LAK cells and from mouse cytolytic T cells. The NH2-terminal amino acid sequence of the polypeptide was found to be identical to calreticulin. Calreticulin is a calcium storage protein and carries a COOH-terminal KDEL sequence, known to act as a retention signal for proteins destined to the lumen of the endoplasmic reticulum. In CTLs, however, calreticulin colocalizes with the lytic perforin to the lysosome-like secretory granules, as confirmed by double label immunofluorescence confocal microscopy. Moreover, when the release of granule-associated proteins was triggered by stimulation of the T cell receptor complex, calreticulin was released along with granzymes A and D. Since perforin is activated and becomes lytic in the presence of calcium, we propose that the role of calreticulin is to prevent organelle autolysis due to the protein's calcium chelator capacity. PMID- 8418195 TI - Function of B cells expressing a human immunoglobulin M rheumatoid factor autoantibody in transgenic mice. AB - We have generated transgenic mice that express the immunoglobulin (Ig)M heavy chain and kappa light chain genes coding for a human IgM rheumatoid factor (RF), Les. Transgenic B cells expressing human IgM RF show striking similarities to their counterparts in normal humans. They comprise a significant proportion of the adult B cell population, but secrete only low levels of RF into the serum. The RF transgene-expressing B cells localize to primary B cell follicles and the mantle zone regions of secondary follicles in the spleen. Using these mice we have been able to show that one of the central functions of normal RF-expressing B cells may be to act as highly efficient antigen-presenting cells for low concentrations of immune-complexed antigen. High levels of secretion of IgM RF can not be induced under normal circumstances, although RF-expressing B cells proliferate well in vitro to both aggregated human IgG and anti-human IgM antibodies. However, these mice are not intrinsically secretion deficient. By crossing the RF transgenic mice with the autoimmune MRL/lpr background, we find a dramatic increase, > 200-fold, in levels of serum RF. The results strongly suggest that a major function of normal resting RF B cells is unrelated to antibody secretion. Rather, the RF B cells in the follicles may play a role in antigen presentation and regulation of immune responses to antibody-bound nonself , and possibly self-antigens. This physiologic role of RF B cells may be disrupted in RF-associated autoimmune disease. PMID- 8418196 TI - Signal transduction in host cells by a glycosylphosphatidylinositol toxin of malaria parasites. AB - In this study, we have identified a dominant glycolipid toxin of Plasmodium falciparum. It is a glycosylphosphatidylinositol (GPI). The parasite GPI moiety, free or associated with protein, induces tumor necrosis factor and interleukin 1 production by macrophages and regulates glucose metabolism in adipocytes. Deacylation with specific phospholipases abolishes cytokine induction, as do inhibitors of protein kinase C. When administered to mice in vivo the parasite GPI induces cytokine release, a transient pyrexia, and hypoglycemia. When administered with sensitizing agents it can elicit a profound and lethal cachexia. Thus, the GPI of Plasmodium is a potent glycolipid toxin that may be responsible for a novel pathogenic process, exerting pleiotropic effects on a variety of host cells by substituting for the endogenous GPI-based second messenger/signal transduction pathways. Antibody to the GPI inhibits these toxic activities, suggesting a rational basis for the development of an antiglycolipid vaccine against malaria. PMID- 8418197 TI - The development of autoimmunity in C57BL/6 lpr mice correlates with the disappearance of natural killer type 1-positive cells: evidence for their suppressive action on bone marrow stem cell proliferation, B cell immunoglobulin secretion, and autoimmune symptoms. AB - F1 hybrid mice are able to acutely reject parental marrow grafts, a phenomenon that is due to natural killer type 1-positive (NK1+) cells. Circumstantial evidence had suggested that the antigenic determinants recognized by these cells are self-antigens, leading to the hypothesis that the physiological role of NK1+ cells is a downregulatory or suppressive function on bone marrow stem cell proliferation and lymphocyte function. In analyzing this hypothesis it is shown here that in young mice there is a temporal correlation between appearance of NK1+ cells in the spleen and the ability to reject allogeneic marrow or to suppress endogenous stem cell proliferation. The reverse situation exists in mice expressing the homozygous lpr gene. Whereas in young mice cells with NK1+ phenotype are demonstrable, these cells disappear with age, i.e., at the time autoimmunity develops. Concomitant with the disappearance of NK1+ cells, the ability to reject marrow grafts and to control endogenous stem cell proliferation also vanishes. The suggestion that the development of autoimmunity is causally related to the disappearance of NK1+ cells is supported by experiments in which NK1+ cells were either eliminated by antibody injection or increased by adoptively transferring cell populations enriched for NK1+ cells into lpr mice. It is shown that removal of cells enhances autoimmunity, whereas injection of NK1+ cells delays the onset of autoimmunity. In vitro assays are presented that demonstrate that suppression of autoantibody-secreting B cells is due to two NK1+ cell populations, one that expresses CD3 and causes specific suppression and one that lacks CD3 and causes nonspecific suppression. PMID- 8418198 TI - Two adjacent residues in staphylococcal enterotoxins A and E determine T cell receptor V beta specificity. AB - The T cell receptor (TCR) V beta-determining region of two bacterial superantigens, staphylococcal enterotoxin A (SEA) and SEE, has been mapped to the COOH-terminal region of SEA and SEE using a panel of recombinant SEA/SEE hybrids. Total TCR V beta mRNA enrichment in human peripheral blood T cell cultures was determined by a novel single-tube amplification technique using a redundant V beta-specific primer. SEA routinely enriched mRNA coding for hV beta 1.1, 5.3, 6.3, 6.4, 6.9, 7.3, 7.4, and 9.1, while SEE, which is 83% homologous to SEA, enriched hV beta 5.1, 6.3, 6.4, 6.9, and 8.1 mRNA. Exchanging residues 206 and 207 was sufficient to convert in toto the TCR V beta response of human peripheral T lymphocytes. In addition, an SEA-reactive murine T cell line, SO3 (mV beta 17), unresponsive to wild-type SEE responded to SEE-S206N207, while an SEE-specific human T cell line, Jurkat (hV beta 8.1), unresponsive to SEA was stimulated strongly by SEA-P206D207. Exchanging all other regions of SEA and SEE except residues 206 and 207 did little to change the V beta response. Thus, the V beta binding region appears to be a stable, discrete domain localized within the COOH terminal region that is largely unaffected by the considerable amino acid variability between SEA and SEE. This region may interact directly with TCR V beta. PMID- 8418199 TI - Putative prethymic T cell precursors within the early human embryonic liver: a molecular and functional analysis. AB - Hematopoietic cells present in the liver in early human fetal life were characterized by phenotypic analysis using a broad panel of monoclonal antibodies. Expression of very late antigen 4 and leukocyte function-associated antigen 3 cell adhesion receptors and 4F2 cell activation molecules was found in all fetal liver hematopoietic cells before acquisition of T cell-, B cell-, or myeloid-specific surface markers, and before the time of intrathymic colonization. Molecular studies showed that expression of the interleukin 2 receptor beta (IL-2R beta) also occurred in the embryonic liver at this early ontogenic stage. In contrast, no expression of IL-2R alpha or IL-2 transcripts was found in fetal liver cells, whereas transcription of the IL-4 gene was detected in a small fetal liver cell subset. Putative T cell precursors were identified among the hematopoietic fetal liver cells by the expression of genes encoding the gamma, delta, epsilon, and zeta invariant chains of the CD3-T cell receptor (TCR) complex. However, no transcription of the polymorphic alpha and beta TCR genes was detected. Functional in vitro assays further demonstrated that fetal liver hematopoietic cells from those early embryos were capable of proliferating in response to T cell growth factors, including IL-4 and IL-2. However, whereas IL-4-induced proliferation paralleled the appearance in vitro of CD45+CD7-CD4dull cells expressing the CD14 myeloid antigen, as well as of CD34+ primitive hematopoietic progenitors, differentiation into CD45+CD7+CD8+CD3- immature T cells was observed when using IL-2. Moreover, coculture with thymic epithelial cell monolayers provided additional evidence that early fetal liver hematopoietic cells may include very primitive T cell precursors, which were able to differentiate in vitro into TCR alpha/beta+ mature T cells. Therefore, our results indicate that, after triggering of the T cell-specific maturation program in primitive fetal liver hematopoietic progenitors, specific signals provided intrathymically by epithelial cells may fulfill the requirements to drive terminal differentiation of prethymically committed T cell precursors. PMID- 8418200 TI - Homozygous scid/scid;beige/beige mice have low levels of spontaneous or neonatal T cell-induced B cell generation. AB - The autosomal recessive scid mutation results in defective immunoglobulin and T cell receptor gene rearrangement. The scid mutation occurred in the allotype congenic C.B-17 line, and up to 25% of C.B-17 scid mice spontaneously produce both T cells and immunoglobulin, a phenotype known as "leaky." Moreover, introduction of neonatal T cells into C.B-17 scid mice leads to immunoglobulin production by 100% of animals. We have produced mice homozygous for both the scid and beige mutations. By contrast with C.B-17 scid mice, BALB/c scid.beige mice have a < 2% incidence of "leakiness." This percentage does not increase with age, and introduction of neonatal T cells fails to rescue immunoglobulin production. This suggests that a gene (or genes) closely linked to the beige locus regulates B and/or T cell development. PMID- 8418201 TI - Tumorigenicity conferred to lymphoma mutant by major histocompatibility complex encoded transporter gene. AB - Presentation of antigenic peptides by major histocompatibility complex (MHC) class I molecules requires MHC-encoded molecules of the adenosine triphosphate binding cassette (ABC) family. Defects in these proteins represent a potential risk, since they are essential links in the machinery of T cell-mediated surveillance which continuously scrutinizes peptide samples of cellular proteins. Nevertheless, transfection of the mouse lymphoma mutant RMA-S with the rat ABC gene mtp2a (homologue to mouse HAM2 and human RING11), commonly termed TAP-2 genes, led to a marked increase in tumor outgrowth potential in vivo. This occurred despite restored antigen presentation and sensitivity to cytotoxic T lymphocytes, and was found to be due to escape from natural killer (NK) cell mediated rejection. It has previously been proposed that adequate expression of self-MHC class I is one important mechanism to avoid elimination by NK cells. Our data argue that a defect in the machinery responsible for processing and loading of peptides into MHC class I molecules is sufficient to render cells sensitive to elimination by NK cells. The latter thus appear to function as a surveillance of the peptide surveillance machinery. PMID- 8418202 TI - Very late antigen 4-dependent adhesion and costimulation of resting human T cells by the bacterial beta 1 integrin ligand invasin. AB - Bacteria and viruses often use the normal biological properties of host adhesion molecules to infect relevant host cells. The outer membrane bacterial protein invasin mediates the attachment of Yersinia pseudotuberculosis to human cells. In vitro studies have shown that four members of the very late antigen (VLA) integrin family of adhesion molecules, VLA-3, VLA-4, VLA-5, and VLA-6, can bind to invasin. Since CD4+ T cells express and use these integrins, we have investigated the interaction of CD4+ T cells with purified invasin. Although VLA integrin-mediated adhesion of T cells to other ligands such as fibronectin does not occur at high levels unless the T cells are activated, resting T cells bind strongly to purified invasin. The binding of resting T cells to invasin requires metabolic activity and an intact cytoskeleton. Although CD4+ T cells express VLA 3, VLA-4, VLA-5, and VLA-6, monoclonal antibody (mAb) blocking studies implicate only VLA-4 as a T cell invasin receptor. Like other integrin ligands, invasin can facilitate T cell proliferative responses induced by a CD3-specific mAb. These results suggest that the nature of the integrin ligand is a critical additional factor that regulates T cell integrin activity, and that direct interactions of T cells with bacterial pathogens such as Yersinia may be relevant to host immune responses to bacterial infection. PMID- 8418203 TI - Reversal of acute and chronic synovial inflammation by anti-transforming growth factor beta. AB - Transforming growth factor beta (TGF-beta) induces leukocyte recruitment and activation, events central to an inflammatory response. In this study, we demonstrate that antagonism of TGF-beta with a neutralizing antibody not only blocks inflammatory cell accumulation, but also tissue pathology in an experimental model of chronic erosive polyarthritis. Intraarticular injection of monoclonal antibody 1D11.16, which inhibits both TGF-beta 1 and TGF-beta 2 bioactivity, into animals receiving an arthropathic dose of bacterial cell walls significantly inhibits arthritis. Inhibition was observed with a single injection of 50 micrograms antibody, and a 1-mg injection blocked acute inflammation > 75% compared with the contralateral joints injected with an irrelevant isotype control antibody (MOPC21) as quantitated by an articular index (AI = 0.93 +/- 0.23 for 1D11.16, and AI = 4.0 +/- 0 on day 4; p < 0.001). Moreover, suppression of the acute arthritis achieved with a single injection of antibody was sustained into the chronic, destructive phase of the disease (on day 18, AI = 0.93 +/- 0.07 vs. AI = 2.6 +/- 0.5; p < 0.01). The decreased inflammatory index associated with anti-TGF-beta treatment was consistent with histopathologic and radiologic evidence of a therapeutic response. These data implicate TGF-beta as a profound agonist not only in the early events responsible for synovial inflammation, but also in the chronicity of streptococcal cell wall fragment-induced inflammation culminating in destructive pathology. Interrupting the cycle of leukocyte recruitment and activation with TGF-beta antagonists may provide a mechanism for resolution of chronic destructive lesions. PMID- 8418204 TI - Heat-shock proteins protect cells from monocyte cytotoxicity: possible mechanism of self-protection. AB - We have previously shown that major heat-shock protein (hsp 70) protects WEHI-S tumor cells from cytotoxicity mediated by tumor necrosis factor alpha (TNF-alpha) and TNF-beta. In the present study, the effect of altered expression of hsp70 and low molecular weight heat-shock protein, hsp27, on tumor cell sensitivity to monocytes and lymphokine-activated killer (LAK) cells was studied. Constitutive and stable expression of transfected human hsp70 rendered cells almost completely resistant to monocytes. Conversely, inhibition of endogenous hsp70 by expression of antisense hsp70 RNA enhanced the sensitivity of cells to monocyte-mediated killing. Surprisingly, overexpression of human hsp27, which does not protect WEHI S cells from TNF killing, conferred partial resistance to monocytes. Only approximately 60% of monocyte-mediated killing of WEHI-S cells could be blocked by neutralizing TNF-alpha antibody or immunoglobulin G-TNF receptor chimeric protein, suggesting the presence of both TNF-dependent and TNF-independent lytic mechanisms. As free radicals have been suggested to be mediators of monocyte cytotoxicity, we tested the sensitivity of transfected cells to oxidative stress. Overexpression of either hsp70 or hsp27 rendered cells partially resistant to hydrogen peroxide. No significant changes in the susceptibility of cell lines overexpressing hsp70 or hsp27 to cytotoxicity mediated by LAK cells were observed. Interestingly, monocytes but not LAK cells contained detectable levels of hsp27 and hsp70 in nonstressed conditions. Taken together, these data indicate that hsp70 protects tumor cells from TNF-mediated monocyte cytotoxicity and that both hsp27 and hsp70 confer resistance to TNF-independent, probably free radical mediated lysis by monocytes. Moreover, hsp27 and hsp70 may provide monocytes with a protective mechanism against their own toxicity. PMID- 8418205 TI - Delayed hematopoietic development in osteopetrotic (op/op) mice. AB - Changes in structure, cellularity, hematopoietic progenitor cell and macrophage content, and osteoclast activity were investigated in the hematopoietic organs of the colony-stimulating factor 1(CSF-1)-less osteopetrotic (op/op) mouse. The data indicated that op/op mice undergo an age-related hematopoietic recovery and resolution of osteopetrosis, suggesting that the hematopoietic system has the capacity to use alternative mechanisms to compensate for the absence of an important multifunctional growth factor, CSF-1. In young animals, op/op femurs were heavily infiltrated with bone, and marrow cellularity was significantly reduced. After 6 wk of age, there was an increase in the marrow space available for hematopoiesis. The femoral cavity of op/op mice progressively enlarged, and by 22 wk of age its appearance and marrow cellularity was comparable to that of controls. The percentage of op/op mononuclear phagocytes, defined by F4/80 antigen expression, progressively increased to normal levels by 35 wk of age. There was no difference in the incidence of both primitive and mononuclear phagocyte-committed, CSF-1-responsive progenitor cells in op/op marrow, but their femoral content was significantly reduced in young mice. During the period of reduced hematopoiesis in the marrow of young op/op mice, splenic hematopoietic activity was elevated. This mutant mouse represents a system for the study of the CSF-1-independent regulatory mechanisms involved in hematopoietic regulation. PMID- 8418206 TI - Human neutrophils express immunoglobulin E (IgE)-binding proteins (Mac-2/epsilon BP) of the S-type lectin family: role in IgE-dependent activation. AB - It has been suggested that neutrophils may be involved in the late-phase reaction of immunoglobulin E (IgE)-dependent hypersensitivity states. However, the identity of neutrophil-associated molecules inducing the release of mediators remains unclear. In this report, we demonstrate that human neutrophils from normal donors or from patients with inflammatory disorders could bind myeloma IgE proteins, especially after desialylation. Northern blot, immunoprecipitation, and flow cytometry analyses revealed that neutrophils did not express Fc epsilon RII/CD23, but rather Mac-2/epsilon binding protein (BP), belonging to the S-type lectin family. Similarly to IgA used as positive control, myeloma IgE proteins, as well as polyclonal IgE antibodies with or without antibody specificity, were both capable of inducing a neutrophil respiratory burst. Anti-Mac-2 but not anti CD23 mAb strongly decreased the IgE-dependent activation of neutrophils, induced either by the specific antigen or by anti-IgE antibodies. These findings open new perspectives on the functional role of neutrophils in IgE-associated diseases including allergic states or parasitic infections. PMID- 8418207 TI - Major histocompatibility complex-specific prolongation of murine skin and cardiac allograft survival after in vivo depletion of V beta+ T cells. AB - The preferential usage of certain T cell receptor (TCR) V beta genes has been well established in several major histocompatibility complex (MHC)-restricted immune responses. However, V beta usage among allogeneic responses remains unclear. Because recent findings of ours and others indicate that V beta 8 predominates in certain Ld-restricted, peptide-specific responses, we examined the V beta 8 usage in allogeneic responses to Ld. To selectively recognize the Ld molecule, cells from BALB/c-H-2dm2 (dm2), the Ld-loss mutant mouse, were stimulated in vitro or in vivo with wild-type BALB/c cells. We report here that after the intraperitoneal administration of the anti-V beta 8 monoclonal antibody (mAb) F23.1, peripheral V beta 8 T cells were depleted from dm2 mice. This in vivo depletion abrogated the ability of dm2 splenocytes to mount a primary response to Ld molecules. This abrogation was specific, since the response of V beta 8-depleted dm2 cells to Kb/Db antigens was the same as that of control nondepleted dm2 cells. Furthermore, in vivo depletion of V beta 8 cells was found to cause a dramatic prolongation of Ld-disparate skin grafts (mean survival time [MST] 22.1 +/- 2.1 vs. 10.3 +/- 1.1 d for saline-treated controls, or 10.9 +/- 1.7 d for controls treated with mAb KJ23 to V beta 17). By contrast, V beta 8 depletion had no effect on recipients grafted with haplotype-mismatched skin or single Dk-locus-disparate skin. These findings demonstrate that V beta 8+ T cells predominate in allogeneic response to Ld but not other alloantigens. The effect of V beta 8 depletion was found to be even more dramatic on recipients grafted with Ld-disparate vascularized heart transplants (MST > 100 vs. 8.6 +/- 0.5 d for controls). In total, these findings establish the efficacy of using mAb to the V beta gene family to specifically and significantly enhance the survival of allografts. The implications of detecting V beta 8 usage in both alloreactive or MHC-restricted TCR responses to the same class I molecule are discussed. PMID- 8418208 TI - Immunoglobulin D (IgD)-deficient mice reveal an auxiliary receptor function for IgD in antigen-mediated recruitment of B cells. AB - To assess the role of immunoglobulin D (IgD) in vivo we generated IgD-deficient mice by gene targeting and studied B cell development and function in the absence of IgD expression. In the mutant animals, conventional and CD5-positive (B1) B cells are present in normal numbers, and the expression of the surface markers CD22 and CD23 in the compartment of conventional B cells indicates acquisition of a mature phenotype. As in wild-type animals, most of the peripheral B cells are resting cells. The IgD-deficient mice respond well to T cell-independent and dependent antigens. However, in heterozygous mutant animals, B cells expressing the wild type IgH locus are overrepresented in the peripheral B cell pool, and T cell-dependent IgG1 responses are further dominated by B cells expressing the wild-type allele. Similarly, in homozygous mutant (IgD-deficient) animals, affinity maturation is delayed in the early primary response compared to control animals, although the mutants are capable of generating high affinity B cell memory. Thus, rather than being involved in major regulatory processes as had been suggested, IgD seems to function as an antigen receptor optimized for efficient recruitment of B cells into antigen-driven responses. The IgD-mediated acceleration of affinity maturation in the early phase of the T cell-dependent primary response may confer to the animal a critical advantage in the defense against pathogens. PMID- 8418209 TI - Conventional B cells, not B-1 cells, are responsible for producing autoantibodies in lpr mice. AB - Mice homozygous for the lpr gene develop autoantibodies and polyclonal B cell activation similar to what is seen in human systemic lupus erythematosus patients. We have previously shown that an lpr-specific intrinsic B cell defect was necessary for autoantibody production in this model. In the current study, we have further defined these autoantibody-producing B cells. Two major subsets of B cells have been described. B-1 cells (CD5+ B cells) can be distinguished from conventional B cells on the basis of phenotype, cytokine secretion, gene expression, anatomical location, and function. In addition, B-1 cells have been implicated in autoimmunity in several murine and human studies. To address the question of which B cell subset produces autoantibodies in lpr mice, we used immunoglobulin heavy chain (Igh) allotype-marked peritoneal (B-1 cell source) and bone marrow (conventional B cell source) cells from lpr mice to establish B cell chimeras. We used two general approaches. In one, we reconstituted sublethally irradiated mice with B-1 cells of one allotype and bone marrow cells of another allotype. In the second method, we suppressed endogenous B cells in neonatal mice with allotype-specific anti-IgM antibody, and injected peritoneal cells of another allotype. After antibody treatment was stopped, the mouse's conventional B cells recovered, but the B-1 subset was only reconstituted by the donor. In both types of chimeras, antichromatin, rheumatoid factor, and anti-single stranded DNA (ssDNA) autoantibodies were produced by the conventional B cell bone marrow source. In addition, an age-related decrease in peritoneal B-1 cells was seen, even in unmanipulated lpr mice. These data show that lpr B-1 cells are not important producers of autoantibodies. Conventional B cells are the source of autoantibodies directed at chromatin, ssDNA, and IgG. PMID- 8418210 TI - Site-specific alterations in the B oligomer that affect receptor-binding activities and mitogenicity of pertussis toxin. AB - Pertussis toxin plays a major role in the pathogenesis of whooping cough and is considered an important constituent of vaccines against this disease. It is composed of five different subunits associated in a molar ratio 1S1:1S2:1S3:2S4:1S5. The S1 subunit is responsible for the ADP-ribosyltransferase activity of the toxin. The B moiety, composed of S2 through S5, recognizes and binds to the target cell receptors and has some ADP-ribosyltransferase independent activities such as mitogenicity. Site-directed mutagenesis of subunits S2 and S3 allowed us to identify amino acid residues involved in receptor binding. Of all the modifications generated, the deletion of Asn 105 in S2 and of Lys 105 in S3 resulted in the more drastic reduction of binding to haptoglobin and CHO cells, respectively. A holotoxin carrying both deletions presented a mitogenicity reduced to an undetectable level. The combination of these B oligomer mutations with two substitutions in the S1 subunit led to the production of a toxin analog with reduced ADP-ribosyltransferase-dependent and independent activities including mitogenicity. As shown by immunoprecipitation with various monoclonal antibodies, the mutant holotoxin was correctly assembled and antigenically similar to the native toxin. This toxin analog induced toxin neutralizing antibodies at the same level as the holotoxin carrying only mutations in the S1 subunit, and may therefore be considered a useful candidate for the development of a new generation vaccine against whooping cough. PMID- 8418212 TI - The major histocompatibility complex-restricted response of recombinant inbred strains of mice to natural tick transmission of Borrelia burgdorferi. AB - The causative agent of Lyme disease, Borrelia burgdorferi, is transmitted by ticks of the Ixodes ricinus complex. In this study, we report the antibody response of recombinant inbred strains of mice of the H-2, b, d, and k haplotypes, infected with B. burgdorferi as a result of exposure to infected I. dammini. The patterns of antibody response assayed by Western blot analysis indicate significant major histocompatibility complex (MHC) restriction to bacterial antigens within the first 2 mo of infection in mice. Other bacterial antigens induce a significant response across the MHC haplotypes tested when assayed on the same bacterial strain used to transmit the infection, but do not crossreact with the same proteins derived from heterologous strains of B. burgdorferi. No response to outer surface protein A was detected at any time during the 60-d period we analyzed this infection. A third group of bacterial antigens appear to generate a MHC-nonrestricted response, and this lack of restriction is maintained when assaying the crossreactivity of the response with other strains of B. burgdorferi. These proteins may provide more accurate diagnostic probes than those currently in use. Finally, there appears to be a significant difference in the expression of most bacterial antigens when the spirochete is cultured for many passages since the same strain of bacterium isolated from low-passage and high-passage preparations exhibit different banding patterns in Western blots when assayed with the same sera. PMID- 8418211 TI - Assessment of ability of murine and human anti-lipid A monoclonal antibodies to bind and neutralize lipopolysaccharide. AB - The use of monoclonal antibodies (mAbs) directed to lipid A for the therapy of gram-negative sepsis is controversial. In an attempt to understand their biologic basis of action, we used a fluid-phase radioimmunoassay to measure binding between bacterial lipopolysaccharide (LPS) and two IgM mAbs directed to lipid A that are being evaluated for the treatment of gram-negative bacterial sepsis. Both antibodies bound 3H-LPS prepared from multiple strains of gram-negative bacteria when large excesses of antibody were used, although binding was modest and only slightly greater than control preparations. We also studied the ability of each anti-lipid A antibody to neutralize some of the biological effects of LPS in vitro. Despite large molar excesses, neither antibody neutralized LPS as assessed by the limulus lysate test, by a mitogenic assay for murine splenocytes, or by the production of cytokines interleukin (IL)-1, IL-6, or tumor necrosis factor from human monocytes in culture medium or in whole blood. Our experiments do not support the hypothesis that either of these anti-lipid A mAbs function by neutralizing the toxic effects of LPS. PMID- 8418214 TI - Behavioral and computational models of spatial attention. AB - Pigeons pecked for food in a spatially cued choice reaction time (RT) task. A brief (50-ms) white light appeared on a left or right key and probabilistically predicted the location (on either the left or right key) of a subsequent target stimulus. The time between cue and target onset (stimulus onset asynchrony), the base rate of left cues, and the probability that the cue correctly predicted the target (cue validity) were experimentally varied. The mean RT to respond to the target key was faster on correctly cued trials (defining a validity effect), decreased for both valid and invalid trials as stimulus onset asynchrony increased (defining an alerting effect), showed a variety of base-rate effects, and did not depend on cue validity. It is shown with a computational-processing model that dynamic interactions of short-term and associative memory processes are sufficient to produce these attentionlike empirical phenomena. PMID- 8418213 TI - Molecular evolution of the human immunoglobulin E response: high incidence of shared mutations and clonal relatedness among epsilon VH5 transcripts from three unrelated patients with atopic dermatitis. AB - We have analyzed the nucleotide sequences of 19 epsilon VH5 transcripts derived from in vivo isotype switched peripheral blood B cells of three patients with atopic dermatitis. Comparison with the patients' own germline VH5 gene segments revealed that the epsilon transcripts were derived from both functional members of the human VH5 gene family and harbored numerous somatic mutations (range 5-36 per VH5 gene). In two patients, we detected clonally related but diverged transcripts, permitting the construction of a genealogical tree in one patient. We observed a high proportion of shared silent (S) and replacement (R) mutations among epsilon VH5 sequences derived from all three individuals, even among transcripts descending from the two different germline VH5 gene segments. A remarkably high number of these mutations is shared with previously reported VH5 genes encoding antibodies with defined specificities. The shared S mutations, and likely a fraction of the R mutations, appear to mark preferential sites ("hot spots") of somatic hypermutations in human VH5 genes. The distribution of R and S mutations over complementarity determining region and framework regions in the majority of VH regions deviated from that characteristic of antigen-driven immune response. We hypothesize that the V regions of immunoglobulin E-bearing B cells have accumulated "selectively neutral" mutations over extended periods of clonal expansion, resulting in unusual R/S ratios. We propose that the molecular characteristics of the epsilon VH regions in atopic dermatitis may be representative of antigens that recurrently or chronically stimulate the immune system. PMID- 8418215 TI - Sexual approach conditioning: unconditioned stimulus factors. AB - Unconditioned stimulus (US) factors were investigated in a Pavlovian sexual conditioning paradigm with male Japanese quail (Coturnix coturnix japonica). Copulation with a female quail was more effective than exposure to a female without copulation, but the latter also produced conditioned responding (Experiment 1). The greater effectiveness of copulatory opportunity as a US was probably not due to nonassociative effects of copulation (Experiment 2). Visual cues of the female, as presented on a taxidermic model, were not effective unless the subjects had prior sexual experience that increased their response to the model (Experiments 3a and 3b). Successful conditioning with noncopulatory female exposure is significant because it allows for sexual learning to occur in a broader range of circumstances than does conditioning with copulation and because it permits conducting multiple trials per day because males do not become satiated as rapidly. PMID- 8418216 TI - Sexual approach conditioning: tests of unconditioned stimulus devaluation using hormone manipulations. AB - Contents of learning that result from conditioned-unconditioned stimulus pairings in sexual approach conditioning were explored with male Japanese quail (Coturnix coturnix japonica). Sexual motivation of subjects conditioned to approach an arbitrary stimulus in a Pavlovian sexual conditioning paradigm was reduced by exposing them to a short photoperiod. Decreased sexual motivation resulted in a decline in sexually conditioned approach behavior (Experiments 1 and 2). Responding was restored when subjects were returned to a long photoperiod (Experiment 1) and when exogenous testosterone was administered (Experiment 2). Decreased sexual motivation did not affect food-conditioned approach behavior (Experiment 3). These results suggest that sexually conditioned approach behavior is mediated by a representation of the unconditioned stimulus, which is activated by the conditioned stimulus. PMID- 8418217 TI - Varieties of learning and memory in animals. AB - It is often assumed that there is more than one kind of learning--or more than one memory system--each of which is specialized for a different function. Yet, the criteria by which the varieties of learning and memory should be distinguished are seldom clear. Learning and memory phenomena can differ from one another across species or situations (and thus be specialized) in a number of different ways. What is needed is a consistent theoretical approach to the whole range of learning phenomena, and one is explored here. Parallels and contrasts in the study of sensory systems illustrate one way to integrate the study of general mechanisms with an appreciation of species-specific adaptations. PMID- 8418218 TI - A retrieval cue for extinction attenuates spontaneous recovery. AB - Four experiments with rats in an appetitive conditioned magazine entry preparation examined spontaneous recovery after extinction. Spontaneous recovery was obtained 6 days but not 5 hr following extinction; recovery depended on the passage of time but not on the removal of a cue that was featured in extinction or on the reintroduction of early-session cues. A cue featured in extinction attenuated recovery when presented on the test. The attenuation effect depended on the cue's correlation with extinction; a cue featured in conditioning did not attenuate recovery. The extinction cue did not evoke responding on its own, suggesting that it was not a conditioned excitor. Retardation tests and a summation test did not reveal that it was a conditioned inhibitor. The cue might work by retrieving a memory of extinction. Spontaneous recovery thus occurs because the subject fails to retrieve an extinction memory. Other accounts of spontaneous recovery are discussed. PMID- 8418219 TI - Negative priming in associative learning: evidence from a serial-conditioning procedure. AB - Three experiments investigated the suggestion that a predicted or primed stimulus commands less processing and consequently elicits a weaker conditioned response (CR) than a stimulus that is not primed. In each experiment rats received initial training in which the presentation of each of 2 serial compounds, A-X and B-Y, was followed by the delivery of food. Subsequently, X's capacity to elicit the CR, approaching the site of food delivery, was assessed when X was preceded by Stimulus A (i.e., primed) or was presented after Stimulus B. Stimulus X elicited a more vigorous response when it was presented after B than when it followed A. These results show that the ability of one event to elicit its CR is reduced if its presentation has been predicted by some other event. This negative priming effect supports one aspect of Wagner's (1981) model of Pavlovian conditioning. PMID- 8418220 TI - Myelosuppression and "conventional" chemotherapy: what price, what benefit? PMID- 8418221 TI - Efficacy and toxicity of multiagent chemotherapy and low-dose involved-field radiotherapy in children and adolescents with Hodgkin's disease. AB - PURPOSE: Between May 1980 and September 1990, 85 patients with Hodgkin's disease were treated with a regimen designed to increase cure rates while reducing late toxicity. PATIENTS AND METHODS: Therapy consisted of five cycles of cyclophosphamide, Oncovin (vincristine; Eli Lilly and Co, Indianapolis, IN), and procarbazine (COP), alternated with four cycles of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) and low-dose (20 Gy) regional radiotherapy. Vincristine and cyclophosphamide were administered as tolerated during irradiation and during the 2- to 4-week rest period between radiation volumes. The need for staging laparotomy was defined by clinical presentation. RESULTS: The median age at diagnosis was 14 years (range, 4 to 20), and 56% of patients were male. The majority (67%) had stage III or IV disease and 68% (19 of 28) of stage II patients had bulky mediastinal disease. Nodular sclerosing histology predominated (67%). Ninety-three percent of patients were alive without disease with a median follow-up of 4.1 years. Abnormalities were detected on chest roentgenograms and/or pulmonary function tests in 58% and 25% of clinically asymptomatic patients who were tested at least 1 year after completion of therapy. The only symptomatic patient had pulmonary fibrosis after treatment with bleomycin (20 U/m2) and mantle (20 Gy)/lung (13 Gy) irradiation, and developed multiple spontaneous pneumothoraces that required cortical stripping. One patient had congestive heart failure 19 months post-treatment, and two had abnormalities on echocardiograms. Thyroid abnormalities occurred in 21 (27%) patients who were assessable for late toxicity. The majority of female patients have had regular menstrual cycles. Six developed ovarian failure, and 10 have had a total of 17 pregnancies. Other than one documented case of oligospermia, information was not available on male fertility. CONCLUSION: The results demonstrate excellent disease control for the COP/ABVD regimen, with acceptable toxicity. PMID- 8418222 TI - Fludarabine potentiates metabolism of cytarabine in patients with acute myelogenous leukemia during therapy. AB - PURPOSE: A protocol was designed to test the hypothesis that fludarabine infusion before arabinosylcytosine (cytarabine [ara-C]) would increase the accumulation of the active metabolite ara-C triphosphate (ara-CTP) in acute myelogenous leukemia (AML) blasts during therapy. PATIENTS AND METHODS: Patients (n = 5) received 1 g/m2 of ara-C infused intravenously (IV) for 2 hours, followed at 20 hours by 30 mg/m2 of fludarabine for 30 minutes. At 24 hours, another identical dose of ara-C was infused. To determine the optimal duration of ara-C infusion following fludarabine, five additional patients were treated on an amended protocol in which the ara-C infusion was extended to 3 g/m2 infused over 6 hours. RESULTS: Comparison of ara-CTP pharmacokinetics in circulating AML cells demonstrated that the area under the curve (AUC) of ara-CTP increased significantly (median, 1.8 fold; range, 1.6 to 2.4; P = .004) after fludarabine infusion. Neither the median plasma ara-C concentrations, the levels of its deamination product arabinosyluracil, nor the rate of ara-CTP elimination from circulating blasts was affected by fludarabine infusion. However, the rate of ara-CTP accumulation by AML cells was increased by a median of 2.0-fold (range, 1.8 to 2.2; P = .001) after fludarabine; the peak occurred within 1 hour of the end of the infusion. In vitro incubation of these cells with arabinosyl-2-fluoroadenine (F-ara-A) before ara-C also produced a median 1.7-fold increase in the ara-CTP accumulation rate. Pharmacology studies in patients receiving 6-hour infusions of ara-C demonstrated that the rate of ara-CTP accumulation was potentiated beyond 2 hours, but not for 6 hours. CONCLUSION: Infusion of fludarabine before ara-C augments the rate of ara-CTP synthesis in circulating AML blasts during therapy. Evaluation of 6-hour ara-C infusions demonstrated that potentiation of ara-CTP synthesis is maximal up to 4 hours in most patients; this pharmacologically optimized regimen should be considered for combination with other antileukemia drugs. PMID- 8418223 TI - Dose-related immunologic effects of levamisole in patients with cancer. AB - PURPOSE: This phase I study was conducted to determine the maximum-tolerated dose (MTD) and the immunologic properties of levamisole in cancer patients when administered alone and in combination with interferon gamma (IFN-gamma). PATIENTS AND METHODS: Twenty patients with advanced cancer and 36 patients with completely resected melanoma (n = 33) or renal cell cancer (n = 3) received levamisole orally every other day for six doses at 1.0, 2.5, 5.0, or 10.0 mg/kg. Ten days later, patients restarted levamisole and began IFN-gamma 0.1 mg/m2 by subcutaneous injection every other day. Blood samples were collected for measurement of neopterin and soluble interleukin-2 receptor (sIL-2R), and for flow-cytometric analysis. RESULTS: The MTD of levamisole was 5 mg/kg, and this was not changed by the addition of IFN-gamma. Dose-related increases in serum levels of neopterin and sIL-2R were noted. Multiple doses of > or = 5 mg/kg of levamisole were required to elicit immune changes, which were more prominent in patients with minimal tumor burdens. Increased expression of CD64 and class I and class II major histocompatibility antigens on monocytes was also observed. The combination of IFN-gamma and levamisole did not result in greater immunologic effects than those observed in previous trials of IFN-gamma alone. CONCLUSION: Levamisole induces dose-related immunologic changes in patients with large or minimal tumor burdens. These changes may be involved in the beneficial effects noted in recent adjuvant trials of levamisole. Ongoing clinical trials should correlate immune changes with response, and trials exploring different schedules of administration using higher, more immunologically active, doses of levamisole should be performed. PMID- 8418224 TI - Infectious complications associated with interleukin-2 administration: a retrospective review of 935 treatment courses. AB - PURPOSE: To determine if interleukin-2 (IL-2)-treated patients are prone to develop clinically significant infections, a retrospective review of 519 patients who received 935 treatment courses over a 38-month period was conducted. MATERIALS AND METHODS: Treatment records of patients receiving intravenous (IV) bolus IL-2 were reviewed. Clinically significant infectious episodes were identified by retrieving data on antibiotic usage and cross-referencing this with microbiology records and chart review. RESULTS: One hundred thirty-nine documented infectious episodes occurred in 122 treatment courses (13.0%); 11 courses were associated with more than one episode of infection. Predominantly urinary tract infections (6.8%) and infections related to IV catheters (5.3%) were encountered. Fifty-eight percent of the catheter-related infections were associated with bacteremia. Other infections included respiratory tract infections (1.0%), skin/muscle infections (0.9%), and miscellaneous infections (0.9%). Bacteria were isolated from the majority of infections. Almost all patients were successfully treated for their infection, with only two septic deaths (0.2%). No difference was noted in infected versus non-infected patients with regard to diagnosis or previous therapy. There was a significant tendency for those patients who developed infection to be older (P2 = .002, Mantel test for trend). Risk factors for the development of infection included vascular access catheters, open wounds, biliary obstruction, or incomplete treatment of previous infections. Over the 3-year study period, the incidence of infection declined from 23% to 7% (P2 < .0001, Mantel test for trend) due to rigorous patient screening, vigilant monitoring for infection, liberal use of antibiotics for suspected infection, and use of prophylactic antibiotics for central venous catheter placement. CONCLUSION: Although treatment with IL-2 may be associated with a slightly increased incidence of bacterial infections, these infections can be successfully managed in the great majority of cases. PMID- 8418225 TI - Octreotide versus loperamide in the treatment of fluorouracil-induced diarrhea: a randomized trial. AB - PURPOSE: Diarrhea is a prominent feature of fluorouracil (5FU) gastrointestinal toxicity, especially when 5FU is combined with leucovorin (LV) or interferon (IFN). No treatment for this condition has been well defined, although drugs, such as diphenoxylate or loperamide, generally are used. The efficacy of octreotide in the treatment of 5FU-induced diarrhea recently has been reported. We performed a randomized trial that compared octreotide with loperamide, the drug most commonly used for therapy for this disorder. PATIENTS AND METHODS: Forty-one patients with grade 2 (four to six stools per day) or grade 3 (seven to nine stools per day; National Cancer Institute toxicity criteria) diarrhea after chemotherapy with a 5FU-containing regimen for colorectal cancer in 28 cases, gastric cancer in six cases, pancreatic cancer in five cases, and breast cancer in two cases, were entered onto the study. Twenty-one patients received octreotide at a dosage of 0.1 mg subcutaneously twice per day for 3 days, and 20 patients received loperamide 4 mg orally initially and then 2 mg every 6 hours for 3 days. The two arms were comparable for age, sex, and primary tumor. Patients were evaluated for response each treatment day; all patients were assessable. RESULTS: Diarrhea resolved in 19 patients in the octreotide arm (one within the first day; four within the second day; and 14 within the third day) versus only three (all after the third day of therapy) in the loperamide arm (P < .005). Median frequency of stools in the 3 days of therapy was four, three, and zero in the octreotide arm and five, five, and five in the loperamide arm. No side effects were observed in both arms. Ten patients on the loperamide arm and only one on the octreotide arm required hospitalization for parenteral replenishment of fluids and electrolytes. CONCLUSION: Octreotide seems to be more effective than loperamide in control of diarrhea and elimination of the need for replenishment of fluids and electrolytes. PMID- 8418226 TI - Granulocyte-macrophage colony-stimulating factor allows safe escalation of dose intensity of chemotherapy in metastatic adult soft tissue sarcomas: a study of the European Organization for Research and Treatment of Cancer Soft Tissue and Bone Sarcoma Group. AB - PURPOSE: This study was designed to test the feasibility of administering doxorubicin at an optimal dose-intensity (> 70 mg/m2 per 21 days) in combination with ifosfamide under recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) cover in patients with metastatic soft tissue sarcomas. PATIENTS AND METHODS: One hundred four eligible patients (of 111 entered) in 16 centers received doxorubicin 75 mg/m2 plus ifosfamide 5 g/m2 every 3 weeks for up to seven cycles. rhGM-CSF (250 micrograms/m2) was administered once or twice daily by subcutaneous injections for up to 14 days between cycles of chemotherapy. RESULTS: Full protocol dose-intensity of chemotherapy was administered to the majority of patients with only 15 of 293 cycles being complicated by febrile episodes that required hospitalization. There were two treatment-related deaths: one from septicemia and one from cardiac failure. The main toxicities attributed to rhGM-CSF were pruritus and rash. A 45% response rate (10% complete remission [CR]) was seen, with a median response duration of 9 months and median survival of 15 months. CONCLUSION: This high-dose regimen of chemotherapy was feasible under rhGM-CSF cover and produced a higher response rate and median survival than previously seen by the European Organization for Research and Treatment of Cancer (EORTC) Soft Tissue Sarcoma Group. A randomized phase III study is now underway comparing this regimen with conventional-dose doxorubicin/ifosfamide to test the dose-response relationship. PMID- 8418227 TI - Body-composition changes in patients who gain weight while receiving megestrol acetate. AB - PURPOSE: Randomized placebo-controlled clinical trials have now established that megestrol acetate causes appetite stimulation and weight gain in patients with anorexia and/or cachexia. There is a paucity of available data to delineate the substance of this increased weight. PATIENTS AND METHODS: Using dual-energy x-ray absorptiometry and tritiated body water methodologies, we performed body composition measurements in 12 patients with advanced cancer before the institution of oral megestrol acetate (800 mg/d) and at subsequent 2-month intervals. RESULTS: Seven of the 12 patients gained weight (2.1 to 16.5 kg) and had repeat body-composition measurements performed at the time of maximum weight gain. The vast majority of the gained weight was clearly from an increase in adipose tissue, while there was a suggestion that an increase in body fluid was responsible for a minority of the weight gain. CONCLUSION: Megestrol acetate induced weight gain is primarily the result of an increase in body mass. PMID- 8418228 TI - A randomized clinical trial comparing melphalan/prednisone with or without interferon alfa-2b in newly diagnosed patients with multiple myeloma: a Cancer and Leukemia Group B study. AB - PURPOSE: This clinical trial was designed to compare the effectiveness of the standard melphalan and prednisone regimen to that of melphalan, prednisone, and interferon in patients with untreated multiple myeloma. PATIENTS AND METHODS: Between October 1985 and March 1988, 278 patients were accrued to a multi institutional, randomized clinical trial. Responding patients were treated for 2 years before termination of therapy. RESULTS: After a median follow-up of 23 months, the overall remission rate for the melphalan/prednisone treatment group was 44% compared with 33% for the group receiving melphalan/prednisone/interferon alfa-2b. The durations of response and survival were identical for the two treatment groups. Median survival was 3.17 years on melphalan/prednisone treatment and 3.0 years on melphalan/prednisone/interferon alfa-2b treatment. Both hematologic and nonhematologic toxicities were greater in the melphalan/prednisone/interferon alfa-2b treatment group, but were usually of a mild or moderate degree and did not interfere with the completion of therapy. The frequency of deaths in the two treatment groups attributable to the treatment itself was similar. CONCLUSION: This study shows no advantage to the concomitant delivery of interferon alfa-2b with standard melphalan and prednisone as initial treatment for patients with multiple myeloma. PMID- 8418229 TI - Phase II trial of mitotane and cisplatin in patients with adrenal carcinoma: a Southwest Oncology Group study. AB - PURPOSE: Previous reports of chemotherapy in patients with adrenal cancer have described responses to cisplatin (CDDP). Because of these reports of good results, a phase II trial that used CDDP with and without mitotane (o,p'DDD) was initiated. PATIENTS AND METHODS: Patients with metastatic or residual adrenocortical carcinoma with objectively measurable disease or biochemical abnormalities were divided into good-risk and poor-risk categories. The latter received CDDP 100 mg/m2 intravenously, and the former received 75 mg/m2. o,p'DDD was administered at a 1,000-mg dose orally four times a day along with cortisone acetate and Florinef (fludrocortisone acetate; Bristol-Myers Squibb Co, Princeton, NJ). RESULTS: Of a total of 42 patients entered onto the study, 37 were eligible. Twenty-nine patients received good-risk and eight received poor risk doses of CDDP. Functioning tumors were present in 45% of patients. Objective responses were noted in 30% (11 of 37) patients (95% confidence interval, 16% to 50%). Response duration was 7.9 months, and the median time to response was 76 days. The median survival of the 37 eligible patients was 11.8 months, and a significant survival advantage was found for patients who underwent prior surgical removal of their primary tumor or bulky disease, who had a performance status of 0 or 1, or who had synchronous metastatic disease. Toxicity of the CDDP and o,p'DDD combination was moderate to severe, and the most common side effects were gastrointestinal, renal, and neurologic. CONCLUSION: The regimen of CDDP and o,p'DDD has activity in patients with adrenocortical carcinoma; however, the toxicity of this treatment was moderate to severe. PMID- 8418230 TI - Preoperative abdominopelvic computed tomographic prediction of optimal cytoreduction in epithelial ovarian carcinoma. AB - PURPOSE: This study was undertaken to assess the ability of computed tomography (CT) to predict the likelihood of optimal primary tumor cytoreduction in women with epithelial ovarian carcinoma. PATIENTS AND METHODS: Fifty-one women with preoperative CT and a histologic diagnosis of epithelial ovarian carcinoma following primary tumor operation by a gynecologic oncologist were identified. Forty-two CT scans were retrospectively analyzed. CT findings of attachment of the omentum to the spleen or disease greater than 2 cm on the diaphragm, liver surface, or parenchyma, pleura, mesentery, gallbladder fossa, or suprarenal paraaortic nodes were coded to represent unresectable disease. CT results were compared with surgical outcome. RESULTS: Twenty-nine of 42 (69%) patients underwent optimal cytoreduction to less than 2 cm residual disease. Successful cytoreduction was accomplished in 23 of 24 patients who fulfilled CT criteria for cytoreduction and six of 18 with CT criteria predictive of inability to perform cytoreduction. CT was highly sensitive for detection of ascites, mesenteric, and omental disease, but was poor for detection of liver involvement, omental attachment to the spleen, gallbladder fossa disease, and peritoneal nodules smaller than 2 cm. The CT findings accurately predicted surgical outcome with a sensitivity of 92.3% and specificity of 79.3%. The positive predictive value was 67% and the negative predictive value was 96%. CONCLUSION: CT scan is an accurate method for the prediction of successful surgical cytoreduction and may have utility in the decision to offer neoadjuvant chemotherapy to certain medically disabled patients, a hypothesis currently under evaluation. PMID- 8418231 TI - Ifosfamide, mesna, and nephrotoxicity in children. AB - PURPOSE: With the increasing use of ifosfamide in pediatric malignancies, nephrotoxicity has emerged as a potentially serious adverse effect, which may be dose-limiting or may cause severe chronic morbidity, including glomerular impairment and/or Fanconi's syndrome. The purpose of this review was (1) to improve the documentation of ifosfamide nephrotoxicity in children, and (2) to consider the possible causative role of ifosfamide metabolites. DESIGN: (1) A grading system was developed that allowed documentation of the nature and severity of published reports of ifosfamide-induced nephrotoxicity, and evaluation of patient and treatment-related risk factors. (2) The relationship between the pharmacology of ifosfamide/mesna and nephrotoxicity was investigated by examination of published data, especially that concerning the quantitative differences in the metabolism of ifosfamide and its nonnephrotoxic structural isomer, cyclophosphamide. RESULTS: (1) Examination of 16 published reports (with assessable data from 40 children) demonstrated that ifosfamide-induced nephrotoxicity was associated with a wide range of patient ages and ifosfamide cumulative doses given by different administration schedules. (2) Chloroacetaldehyde, a major metabolite of ifosfamide only, may be at least partly responsible for the renal toxicity of this drug. Although mesna may be capable of detoxifying the toxic metabolite(s), delivery to the renal tubule may not be sufficient to provide adequate protection of tubular glutathione from depletion by the metabolite(s), which results in a failure to prevent nephrotoxicity. CONCLUSION: Increased understanding of the interindividual variability in the extent and nature of ifosfamide metabolism, which may be a major determinant of susceptibility to renal damage, may lead to improved use of the drug with less nephrotoxicity. PMID- 8418232 TI - Timing of radiotherapy in the treatment of early-stage breast cancer. PMID- 8418233 TI - In rectal carcinoma, colostomy or no colostomy: is this the question? PMID- 8418234 TI - Use of stereotactic radiosurgery in the treatment of malignant glioma. PMID- 8418235 TI - CPT-11-induced cholinergic effects in cancer patients. PMID- 8418236 TI - Ondansetron compatible with sodium acetate. PMID- 8418237 TI - Intensive preoperative chemotherapy with colony-stimulating factor for resectable adenocarcinoma of the esophagus or gastroesophageal junction. AB - PURPOSE: The curative resection rate in patients with potentially resectable carcinoma of the esophagus is approximately 55% and their median survival time is 11 months. Preoperative chemotherapy with high doses of chemotherapeutic agents was used to evaluate clinical and pathologic responses, curative resection rate, toxicity, and survival. Colony-stimulating factor (CSF) was added to reduce the severity of myelosuppression. PATIENTS AND METHODS: Twenty-six consecutive assessable patients with potentially resectable adenocarcinoma of the esophagus or gastroesophageal junction were treated with two preoperative courses of intensive chemotherapy (etoposide, doxorubicin, and cisplatin [EAP]) with granulocyte-macrophage CSF (GM-CSF). Additional three conventional-dose postoperative chemotherapy courses without GM-CSF were given to patients who responded to preoperative chemotherapy. RESULTS: A median of three courses (range, one to six), were administered. Of 27 patients, 26 were assessable for response to preoperative EAP; 13 (50%) achieved a major response. Among 23 patients who underwent surgery, 15 (65%) had a curative resection (58% of 26 assessable patients); none of the patients had a pathologic complete response, but two patients had only microscopic carcinoma in the resected specimen. Six patients had carcinoma present at the resection margins and received postoperative radiotherapy. Two patients were found to have liver metastases at exploration. At a median follow-up of 22 months, the median survival of 26 patients was 12.5 months (range, 2 to 32 +). Fourteen patients died of their carcinoma; two patients died of treatment-related causes; one died of an unrelated CNS arterial malformation; and the causes of death in two patients remain unknown. Seven patients are alive with no evidence of relapse. Major toxicities of this regimen included severe myelosuppression, nausea and vomiting, infections, and severe constitutional symptoms related to GM-CSF. However, subcutaneous injection of GM-CSF was well tolerated. CONCLUSION: High-dose EAP is active against locoregional adenocarcinoma of the esophagus and gastroesophageal junction but can be associated with significant toxicity. Although this strategy remains attractive and needs to be developed further, less toxic and more effective regimens need to be identified. PMID- 8418238 TI - Adjuvant tamoxifen versus placebo in elderly women with node-positive breast cancer: long-term follow-up and causes of death. AB - PURPOSE: This study analyzes the long-term results and causes of death in elderly women with node-positive breast cancer who participated in a double-blind adjuvant trial that compared tamoxifen with placebo to determine the benefit of 2 years of treatment. PATIENTS AND METHODS: One hundred eighty-one women 65 to 84 years old were given 20 mg of tamoxifen or placebo daily for 2 years after stratification by estrogen receptor status, tumor size, and degree of lymph node involvement. Approximately 30% of patients were older than 70 years and 20% were older than 75 years. Eighty-five percent were estrogen receptor-positive. Median follow-up was 10 years. RESULTS: Among the 168 eligible patients, there have been 98 recurrences (59 placebo v 39 tamoxifen), with reduced distant and bone-only first sites in patients treated with tamoxifen. Median time to failure was 4.4 years for placebo versus 7.4 years for tamoxifen (log-rank P = .001). A similar number of new nonbreast cancers occurred in each arm (seven placebo v six tamoxifen), but a reduced number of opposite-breast cancers (five placebo v one tamoxifen) was noted. Overall, there were 102 deaths (57 placebo v 45 tamoxifen). Median survivals were 8.0 years with placebo and 8.5 years with tamoxifen (log rank P = .063); 50% of the tamoxifen patients and 33% of the placebo patients are still alive. Sixty-one percent of the deaths were reported to have been caused by breast cancer recurrence, 4% by other cancers, and 22% by the sequelae of non cancer-related illness, with equal distributions for cardiovascular and cerebrovascular disease. There was no increase in the number of endometrial or other types of cancer, or thrombotic or orthopedic complications in this older group. CONCLUSION: Tamoxifen currently is the treatment of choice for elderly women with breast cancer. It extends the time to treatment failure by 3 years and reduces the number of recurrences, deaths, distant and bone-only first recurrences, and second breast cancers. PMID- 8418239 TI - Age and the treatment of breast cancer. PMID- 8418240 TI - Prognosis following salvage mastectomy for recurrence in the breast after conservative surgery and radiation therapy for early-stage breast cancer. AB - PURPOSE: The prognosis and factors that influence prognosis following salvage mastectomy in patients with recurrence in the treated breast after conservative surgery (CS) and radiation therapy (RT) were investigated. MATERIALS AND METHODS: A total of 1,593 patients with stage I or II invasive breast cancer were treated following gross total excision of the tumor at the Joint Center for Radiation Therapy (JCRT) between 1968 and 1985. One hundred sixty-six of the 1,593 (10%) had subsequent recurrence in the breast. Of these, 123 had salvage mastectomy and constitute the study population. The recurrent tumor was predominantly invasive in 99 patients, noninvasive in 14, and focally invasive in 10. Following mastectomy, chemotherapy or hormonal therapy was administered to 29 patients. The median follow-up time was 39 months after salvage mastectomy. RESULTS: The 5-year actuarial rate of further local or distant relapse for the entire group was 41%. None of the 24 patients with focally invasive or noninvasive tumors had a subsequent relapse. In comparison, the 5-year actuarial rate of further relapse in the 99 patients with a predominantly invasive recurrence was 52% (P = .001). The method of detection of the recurrence, the age of the patient at initial diagnosis, the disease-free interval, and the location of the recurrence in the breast were not found to have a statistically significant association with the risk of further relapse. CONCLUSION: We conclude that the histology of the recurrent tumor is an important prognostic factor for the risk of further relapse. Patients with purely noninvasive or focally invasive tumors have an excellent prognosis following salvage mastectomy. In contrast, patients with predominantly invasive tumors are at substantial risk for further relapse. PMID- 8418242 TI - Karnofsky memorial lecture. Ode to methotrexate. PMID- 8418241 TI - A pilot study of pi-class glutathione S-transferase expression in breast cancer: correlation with estrogen receptor expression and prognosis in node-negative breast cancer. AB - PURPOSE AND METHODS: Previous studies have indicated that RNA levels for pi-class glutathione S-transferase (GST pi), a phase II, drug-metabolizing enzyme, were inversely related to estrogen receptor (ER) and progesterone receptor (PR) levels in human breast tumors. Because GST pi also is expressed in normal breast epithelium, an immunohistochemical assay that uses affinity-purified polyclonal antibodies to GST pi was developed to examine the possible relationship between GST pi expression in breast cancer cells and hormone receptor expression, as well as prognosis, in patients with primary breast cancer. RESULTS: A strong inverse correlation between GST pi expression and ER (two-sided P [P2] = .002) and PR status (P2 = .023) was found in our study of 189 patients with primary breast cancer. GST pi expression was not related to tumor size, nodal metastasis, nuclear grade, histology, or age of the patient. In node-negative breast cancer (n = 72), increased GST pi expression was associated with decreased disease-free survival (DFS) and overall survival (OS). When GST pi expression was divided into categories of negative (no GST pi-positive tumor cells), intermediate (1% to 70% GST pi-positive tumor cells), and high (> 70% GST pi-positive tumor cells), the relative risk of tumor recurrence in patients with node-negative breast cancer was increased 3.39-fold for each successive category of expression (P2 = .0045; 95% confidence interval, 1.46 to 7.87) and the relative risk of death was increased 4.49-fold for each successive category (P2 = .0003; 95% confidence interval, 2.02 to 10.42). The actuarial 5-year OS was 100%, 79%, and 51%, and the DFS was 94%, 77%, and 44%, for the negative, intermediate, and high tumor groups, respectively. Among the factors studied in multivariate analysis (ER status, PR status, nuclear grade, and tumor size), GST pi expression was the factor that most accurately predicted shorter DFS and OS in node-negative patients. CONCLUSION: GST pi expression is inversely related to hormone receptor status in breast cancer. This pilot study also suggests that increased GST pi expression may be an important predictor of early recurrence and death in node-negative breast cancer patients that merits additional investigation. PMID- 8418244 TI - Survival of breast cancer patients receiving adjunctive psychosocial support therapy: a 10-year follow-up study. AB - PURPOSE: The impact of an adjunctive psychosocial support program on length of survival with breast cancer was evaluated in a retrospective cohort study. The duration of observation of survival was extended 10 years beyond a previous study of the same cohort of patients. PATIENTS AND METHODS: One hundred two nonparticipants were individually matched to 34 participants on major prognostic factors. Both groups were monitored from the date of cancer diagnosis (1971 through 1980) until March 1991. The support program consisted of weekly cancer peer support and family therapy, individual counseling, and use of positive mental imagery. Survival analysis controlled for the effects of other major prognostic factors in the outcome of breast cancer. RESULTS: The mean +/- SD survival time from date of cancer diagnosis to last date of follow-up was 96.0 +/ 53.2 months in the participant group compared with 85.1 +/- 63.4 months in the nonparticipant group, a nonsignificant difference (P = .1). Median survival was 84.0 months for participants (95% confidence interval [CI], 59 to 133) and 66.0 months for nonparticipants (95% CI, 48 to 105). A second analysis restricted nonparticipants to those who had a survival time > or = that of the matched case at time of entry into the support program. Survival increased to a mean of 101.1 months (median, 105.0; 95% CI, 71 to 132) for nonparticipants and remained unchanged for participants, also a statistically nonsignificant difference (P = .9). CONCLUSION: While the program may have beneficial effects on quality of life, this study does not indicate a significant favorable impact on survival with breast cancer or that the program is serving as a social locus for the gathering of exceptional survivors. PMID- 8418243 TI - Double-blind controlled trial of oral clodronate in patients with bone metastases from breast cancer. AB - PURPOSE: Osteolytic metastases often give rise to hypercalcemia, fracture, and bone pain, and occur commonly in patients with recurrent breast cancer. We assessed the bisphosphonate, clodronate, which has proven to be a useful treatment for hypercalcemia and may be a potent inhibitor of tumor-induced osteolysis, for its effect on reducing the osseous complications of metastatic breast cancer. PATIENTS AND METHODS: We studied 173 patients with bone metastases due to breast cancer in a randomized, double-blind, placebo-controlled trial of oral clodronate 1,600 mg/d (85 patients) compared with an identical placebo (88 patients). RESULTS: The patients in each wing were comparable in their clinical, radiologic, and biochemical characteristics at trial entry. In patients who received clodronate, there was a significant reduction compared with placebo in the total number of hypercalcemic episodes (28 v 52; P < .01), in the number of terminal hypercalcemic episodes (seven v 17; P < .05), in the incidence of vertebral fractures (84 v 124 per 100 patient-years; P < .025), and in the rate of vertebral deformity (168 v 252 per 100 patient-years; P < .001). The combined rate of all morbid skeletal events was significantly reduced (218.6 v 304.8 per 100 patient-years; P < .001). Trends were seen in favor of clodronate for nonvertebral fracture rates and radiotherapy requirements for bone pain (particularly spinal pain). No significant survival differences and no significant differences in side effects were observed between the two groups. CONCLUSIONS: These findings indicate that oral clodronate has a beneficial effect on the skeletal morbidity associated with breast cancer and should be considered as antiosteolytic therapy in affected patients. It deserves further investigation as an adjuvant therapy in operable breast cancer and in patients with nonosseous recurrence who are at high risk for bone metastases. PMID- 8418245 TI - Multicenter phase II trial of mitoxantrone in patients with advanced nasopharyngeal carcinoma in Southeast Asia: an Asian-Oceanian Clinical Oncology Association Group study. AB - PURPOSE: Patients with advanced nasopharyngeal carcinoma (NPC) have a high incidence of recurrence and often develop distant metastases despite local control. This prospective multicenter phase II trial was conducted to evaluate the safety and efficacy of Novantrone (mitoxantrone; Lederle Laboratories, Wayne, NJ) in the therapy of patients with advanced NPC. PATIENTS AND METHODS: One hundred eight patients with advanced NPC, namely, those with recurrent or persistent disease following primary radiotherapy, or newly diagnosed metastatic disease, were treated with mitoxantrone. Mitoxantrone was administered intravenously at an initial dose of 12 mg/m2 and repeated every 3 weeks, with dose escalation to a maximum of 14 mg/m2. The distribution of histologic subtypes was representative of NPC, with the majority being (61%) undifferentiated (or anaplastic) carcinoma. RESULTS: The overall response rate (complete response [CR] and partial response [PR]) was 25% (95% confidence interval, 17% to 33%). The median response duration, time to treatment failure, and survival duration were 140, 82, and 394 days, respectively. Histology (poorly differentiated squamous cell) was found to be the only important factor in predicting response (P = .04) based on a multivariate analysis of nine pretreatment characteristics. The major dose-limiting toxicity was leukopenia. The incidences of nausea/vomiting, alopecia, and stomatitis/mucositis were 34%, 6%, and 3%, respectively. None were severe. Two patients had asymptomatic, moderate Alexander-grade cardiotoxicity. CONCLUSION: This study represents a large, controlled multicenter trial of single agent mitoxantrone in the treatment of advanced NPC. Mitoxantrone was well tolerated and produced an overall response rate comparable to that of other single-agent therapies used in the treatment of advanced head and neck cancer. Combination trials with mitoxantrone for advanced disease should be considered. PMID- 8418246 TI - Randomized comparison of diaziquone and carmustine in the treatment of adults with anaplastic glioma. AB - PURPOSE: We conducted a phase III trial comparing intravenous (IV) diaziquone (AZQ) and carmustine (BCNU) as single agents in patients with cerebral anaplastic gliomas who had received surgery and radiotherapy. Its purpose was to compare the efficacy of AZQ with that of BCNU, the standard agent for brain tumor chemotherapy. PATIENTS AND METHODS: Randomization between the two regimens occurred 8 weeks after completion of radiotherapy. A total of 251 patients were randomized to receive either AZQ or BCNU, and there were no significant differences between the two treatment arms in any of the known prognostic variables, including age, histologic grade, and Karnofsky performance status (KPS). RESULTS: There was no significant difference in either time to tumor progression or survival between the two treatment arms. Age and histology were strong predictors of outcome, whereas KPS had relatively less effect. Three groups of patients with distinctly different outcomes could be identified: (1) older age (45+) and glioblastoma/gliosarcoma (GBM/GS) patients had a median survival of 37 weeks after randomization; (2) patients with either older age or GBM/GS had a median survival of 61 weeks; and (3) younger age (< 45) and non GBM/GS (usually anaplastic astrocytoma) patients had a median survival of 147 weeks. Toxicity was primarily hematologic, although acute gastrointestinal toxicity and chronic pulmonary toxicity were more common with BCNU. Patients randomized to AZQ who had significant hematologic toxicity that required dose reduction after the first treatment cycle had significantly longer time to tumor progression and survival than those who did not require dose reduction (P = .011 and .016, respectively). CONCLUSION: There was no significant difference in efficacy between AZQ and BCNU in patients with anaplastic gliomas as tested in this study, although AZQ was somewhat better tolerated. PMID- 8418247 TI - Stage III neuroblastoma over 1 year of age at diagnosis: improved survival with intensive multimodality therapy including multiple alkylating agents. AB - PURPOSE: A nonrandomized, single-arm trial was conducted to assess the efficacy of multimodality therapy including intensive chemotherapy with multiple alkylating agents in the treatment of children with Evans stage III neuroblastoma older than 1 year at diagnosis. PATIENTS AND METHODS: Twenty-five patients with a median age of 18 months at diagnosis were treated with multimodality therapy including surgery and chemotherapy using either nitrogen mustard (mechlorethamine), doxorubicin, cisplatin, dacarbazine (DTIC), vincristine, and cyclophosphamide (MADDOC) or cisplatin and cyclophosphamide induction followed by maintenance MADDOC (induction MADDOC) protocols. Sixteen of 25 patients also received radiotherapy to the tumor bed and primary lymph nodes. Event-free survival (EFS) was compared with that reported previously in the literature. N myc amplification was evaluated prospectively and the Shimada classification was evaluated retrospectively as potential prognostic factors. RESULTS: We report a 72% EFS (95% confidence interval +/- 18%) with a median follow-up of 85 months. EFS was significantly worse for patients with tumors demonstrating N-myc amplification (P = .018). Patients classified as favorable according to the Shimada system experienced a significantly better EFS (P = .04), but unfavorable patients still maintained a 60% EFS. CONCLUSION: Intensive multimodality treatment including MADDOC and induction MADDOC chemotherapy provides a very good EFS for children older than 1 year who have stage III neuroblastoma. Children classified as favorable according to the Shimada system have a better prognosis. Patients whose tumors demonstrate N-myc amplification have a poor prognosis despite therapy. PMID- 8418248 TI - Influence of the blood glucose concentration on FDG uptake in cancer--a PET study. AB - Radiolabeled [11F]-2-fluoro-2-deoxy-D-glucose (FDG) is a glucose analogue widely used to study tumor metabolism by means of positron emission tomography (PET). Little is known about the effect of hyperglycemia on FDG uptake and PET imaging of tumors. Five patients with head and neck cancer underwent two PET studies prior to cancer therapy, first in the fasting state and then 2-5 days later after oral glucose loading. FDG uptake was measured with standardized uptake values (SUV) and Ki values according to Patlak et al. The fasting SUVs ranged from 4.1 to 10.9 and Kis from 0.021 min-1 to 0.067 min-1, whereas after loading both the SUVs (range 2.2-5.9, p < 0.02) and Ki values (range 0.006-0.042 min-1, p < 0.05) decreased significantly, and the quality of the PET images became markedly poorer. The FDG metabolic rate (Ki x P-Gluc) remained similar in different plasma glucose concentrations in tumors, but increased clearly in muscles after loading. Therefore, patients entering PET-FDG studies should fast and their blood glucose concentration needs to be taken into account when evaluating FDG accumulation. PMID- 8418249 TI - Uptake and biodistribution of technetium-99m-MD32P during rat tibial bone repair. AB - The present study was carried out in order to test the hypothesis that intravenously injected Tc-MDP separates into its technetium and methylene diphosphonate components in the bone, and that the technetium is preferentially taken-up by the newly-formed osteoid, while the methylene diphosphonate is taken up by the forming mineral. Uptake of Tc-MDP was studied in a rat model of primary bone formation following tibial bone marrow ablation. Each of five radiopharmaceuticals (99mTCO4, 99mTc-MDP, Tc-MD32P, 99mTc-MD32P or MD32P) was injected and their uptake was followed in the whole bone as well as in the organic and inorganic phases of the bone. Irrespective of the radionuclides injected, 99mTc was always taken-up preferentially by the organic phase, while the 32P was preferentially taken-up by the inorganic phase. When 99mTcO4 was injected, it was not taken up by the bone at all. These results indicate that the increased incorporation of 99mTc, when administered as 99mTc-MDP during bone healing, reflects an enhancement in the formation of the organic matrix and not of the calcification process. The study also suggests that the 99mTc-MDP dissociates into its technetium and methylene diphosphonate moieties, which are then adsorbed onto the organic and inorganic phases respectively. PMID- 8418250 TI - Directly and indirectly technetium-99m-labeled antibodies--a comparison of in vitro and animal in vivo properties. AB - To investigate the in vivo and in vitro properties of 99mTc when labeled to antibodies via one direct and one indirect method, the B72.3 and C110 IgG antibodies were radiolabeled directly via stannous ion reduction and indirectly via the hydrazino nicotinamide chelator and compared in vitro and in vivo. Antibody avidity (but not immunoreactive fraction) appeared to be independent of labeling methods for both antibodies. Following stannous ion reduction, antibodies were fragmented by denaturing SDS PAGE although only slight evidence of fragmentation was found in vivo. The direct label was instable to transchelation to cysteine and glutathione in vitro and in vivo. Following intravenous administration, urinary excretion of activity was threefold greater for the direct label and was almost exclusively labeled cysteine and glutathione. Significant differences in the biodistribution of 99mTc were also observed: liver levels were lower, kidney levels were higher and clearance of label from blood and tissues was faster for the direct label. At Day 1, tumor accumulation was threefold lower for the direct label although most normal tissues were also lower. In conclusion, when labeled to two antibodies by one direct method, 99mTc is unstable towards transchelation relative to one indirect method. These relative instabilities greatly influenced the biodistributions in mice and may influence the quality of images obtained in patients. PMID- 8418251 TI - Fluorodeoxyglucose imaging of advanced head and neck cancer after chemotherapy. AB - Positron emission tomography (PET) was applied to evaluate therapeutic effects in patients with advanced head and neck cancer for use in monitoring therapy. In 18 patients with histologically proven head and neck cancer, PET studies with 330 440 MBq 18F-fluorodeoxyglucose (FDG) were performed prior to the first chemotherapeutic cycle with cisplatin and 5-FU. A second examination after the first chemotherapeutic cycle was performed in 11 patients. Tumor or lymph node volumes were determined from CT slices and the growth rate was calculated assuming an exponential function. Uptake in a region of interest was used for the quantitative evaluation of the PET images after standardization to injected dose and body weight. FDG data were available for 6 tumors and 10 metastases, volumetric data for 5 tumors and 7 metastases. One lesion showed an increase, seven a decrease in FDG uptake and eight lesions remained unchanged. Multiple lymph nodes in the same patient showed different baseline metabolisms and also different changes following therapy. Tumors were more sensitive to therapy than lymph node metastases. The growth rate and the change in FDG uptake were highly correlated with different regression functions for tumors and lymph node metastases. These data demonstrate that PET with FDG can be used to assess early chemotherapeutic effects. The information gained with PET can be included for treatment planning in patients undergoing systemic chemotherapy. PMID- 8418252 TI - A PET radiotracer for studying serotonin uptake sites: carbon-11-McN-5652Z. AB - A radioligand for imaging central serotonin (5-hydroxytryptamine; 5-HT) uptake sites by positron emission tomography (PET) has yet to be developed. Such a tracer would be useful for the study of normal and altered serotonergic neurotransmission as well as for the detection of serotonergic neurotoxicity. This paper describes the labeling of the highly potent serotonin (5-HT) uptake blocker, McN-5652-Z (trans-1,2,3,5,6,10 beta-hexahydro-6-[4 (methylthio)phenyl]pyrrolo-[2,1-a]-isoquinoline; racemic mixture), with 11C and the evaluation of this radiotracer in rodents with respect to its in vivo binding characteristics. In mouse brain, 11C-McN-5652-Z accumulated rapidly in regions with high densities of 5-HT uptake sites. The ratio between hypothalamus and cerebellum was 1.5:1 at 15 min and increased with time to 4.6:1 at 90 min after injection. The distribution of 11C-McN-5652 in rat brain at 60 min correlated well with regional concentrations of 5-HT uptake sites (r = 0.86). The specificity and selectivity of 11C-McN-5652 binding to the 5-HT transporter were tested by preinjecting blocking doses of known 5-HT, dopamine and norepinephrine uptake inhibitors, and a 5-HT2 receptor blocker before injection of the radiotracer. Preinjection of increasing doses of unlabeled McN-5652-Z inhibited 11C-McN-5652-Z binding in a dose-dependent fashion. These results suggest that the in vivo binding of the radiotracer was specific, selective for 5-HT uptake sites, saturable and that 11C-McN-5652-Z holds promise as a radiotracer for PET imaging of 5-HT uptake sites in the mammalian brain. PMID- 8418253 TI - Cerebrovascular accident associated with dipyridamole thallium-201 myocardial imaging: case report. AB - A patient with known aortoiliac occlusive disease and hypertension suffered a cerebrovascular accident 6.5 min after the administration of intravenous dipyridamole during a 201Tl myocardial study. Despite aminophylline administration, the patient developed a completed stroke. The mechanism most likely responsible for precipitating this patient's stroke is dipyridamole induced vascular steal. Although dipyridamole-201Tl myocardial imaging is relatively free of major complications, the morbidity and mortality associated with a cerebrovascular accident is significant. The possibility of precipitating a cerebrovascular accident during dipyridamole-201Tl imaging should be considered in all patients with significant risk factors for stroke before performing a dipyridamole cardiac study. PMID- 8418254 TI - PET scans advance as tool in insanity defense. PMID- 8418255 TI - Bile leak from gallbladder perforation mimicking bowel activity and a false negative result in a morphine-augmented cholescintigraphy. AB - Cholescintigraphy of a patient with bile leak demonstrated intra-abdominal activity that mimicked normal bowel activity. Because the gallbladder was not visualized, morphine was injected intravenously. Gallbladder activity after morphine injection was misleading in the finding of chronic cholecystitis. Concurrent abdominal sonography and computerized tomography revealed a thickened gallbladder wall with a gallstone and pericholecystic fluid collection. Exploratory laparotomy confirmed acute and chronic cholecystitis, cholelithiasis, choledocholithiasis, and a pericholecystic abscess. The false-negative conclusion for acute cholecystitis in the patient's morphine-augmented cholescintigraphy resulted from an acceleration of bile leakage due to pre-existing gallbladder perforation. PMID- 8418256 TI - Spontaneous silent myocardial ischemia assessed by technetium-99m-sestamibi imaging. AB - We report on an 80-yr-old patient with a history of inferior myocardial infarction. The patient was injected with 15 mCi of 99mTc-sestamibi when an electrocardiogram revealed new asymptomatic inferolateral ST-segment depressions. A second 99mTc study was performed 72 hr after the initial injection and after ST segment changes had resolved. Scintigraphic acquisitions using SPECT revealed a large reduction in photon activity in the inferolateral and inferoposterior walls with 70%-80% normalization. Therefore, 99mTc-sestamibi imaging is useful in assessing asymptomatic silent myocardial ischemia. PMID- 8418257 TI - A procedure for patient repositioning and compensation for misalignment between transmission and emission data in PET heart studies. AB - A procedure for patient repositioning and compensation for misalignment between transmission and emission data in positron emission tomography (PET) heart studies has been developed. Following the transmission scan (TR1), patients are moved from the scanner bed for the administration of the tracer, and repositioned when ready for the emission scan (EM1). A short postinjection transmission scan (TR2) is performed at the end of the EM1 study. TR1 and TR2 images are compared to recognize misalignment between transmission and emission studies. TR1 sinograms are compensated for misalignment to allow for a proper attenuation correction. The procedure has been tested on phantom and [18F]FDG PET heart studies. Misalignments down to 2.5 mm translation and 1 degree rotation in the transaxial plane and 4 mm in the axial direction can be recognized and compensated for. The procedure is suitable for clinical purposes, allowing reduction of patient time on the scanner bed, increased patient comfort and significant increase of patient throughput. PMID- 8418258 TI - Attenuation correction using count-limited transmission data in positron emission tomography. AB - Poisson noise in transmission data can have a significant influence on the statistical uncertainty of PET measurements, particularly at low transmission count rates. In this paper, we investigate the effect of transmission data processing on noise and quantitative accuracy of reconstructed PET images. Differences in spatial resolution between emission and transmission measurements due to transmission data smoothing are shown to have a significant influence on quantitative accuracy and can lead to artifacts in the reconstructed image. In addition, the noise suppression of this technique is insufficient to greatly reduce transmission scan times. Based on these findings, improved strategies for processing count-limited transmission data have been developed, including a method using segmentation of attenuation images. Using this method, accurate attenuation correction can be performed using transmission scan times as low as 2 min without increasing noise in reconstructed PET images. PMID- 8418259 TI - Routine application of fractionated HMPAO stored at -70 degrees C for WBC scintigraphy. AB - The radiochemical purity of 99mTc-HMPAO prepared from fractions of reconstituted HMPAO stored at -70 degrees C and its application in the radiolabeling of human granulocytes was investigated. Upon reconstitution of a vial of HMPAO with 1 ml of saline and subsequent freezing at -70 degrees C, small fractions were obtained on each of four consecutive days with the vial being refrozen after each dispensing. Following radiolabeling of the HMPAO fractions with pertechnetate, mean radiochemical purity results met or exceeded manufacturers' specifications for the radiopharmaceutical on each of the four days. Imaging with radiolabeled granulocytes using 99mTc-HMPAO prepared by this technique resulted in high quality clinical studies. These results demonstrate that a vial of HMPAO can be fractionated, after storage at -70 degrees C with no loss of clinical utility for radiolabeling granulocytes and considerable cost savings. PMID- 8418260 TI - Medical imaging in the nineties: new directions for nuclear medicine. PMID- 8418261 TI - The new molecular medicine. PMID- 8418262 TI - We are training our future. PMID- 8418263 TI - Thallium scintigraphy in the evaluation of mass abnormalities of the breast. AB - Palpable mass abnormalities of the breast are often difficult to evaluate mammographically, especially in patients with fibrocystic change and dense breasts. The current study evaluates 201TI scintigraphy as a potential test in detecting malignancy and in differentiating malignant from benign masses. Eighty one female patients underwent thallium scintigraphy of the breast because of palpable breast masses. An additional 30 females with no palpable breast abnormalities were also studied using 201TI. Of 44 patients with palpable breast carcinomas, 42 carcinomas (96%) were detected using 201TI scintigraphy. Three of three patients had other primary breast malignancies that were also detected. In contrast, 19 patients with palpable breast abnormalities shown on biopsy to be benign fibrocystic disease processes were not detectable on thallium studies. Of two patients with fat necrosis, none were detectable. Three of 13 patients had adenomas of the breast (23%) that were detected. The three detectable adenomas were all highly cellular. The smallest detectable carcinoma was an adenocarcinoma measuring 1.3 x 1.1 x 0.9 cm. Thallium-201 scintigraphy of palpable breast lesions is an effective test for evaluation of palpable masses. Sensitivity for detection of malignant masses greater than 1.5 cm is high. Highly cellular adenomas, however, may demonstrate significant 201TI uptake. Benign fibrocystic disease is not detectable with thallium scintigraphy. Thallium scintigraphy of breast lesions is an effective means of differentiating benign from malignant lesions. PMID- 8418264 TI - Congress erects ambiguous wall against BRC exemptions. PMID- 8418265 TI - Ultrastructural histology correlates with results of thallium-201/technetium-99m parathyroid subtraction scintigraphy. AB - Specimens from 15 scintigraphically true-positive adenomas (golden standard: histology), 15 false-negative adenomas, 15 true-positive hyperplasias, 15 false negative hyperplasias, 15 true-negative normal glands from patients with hyperparathyroidism, and 15 normal glands from patients without hyperparathyroidism, all selected randomly, were studied. After fixation, sectioning and H and E staining, in all 90 tissues the number of oxyphil, chief, and clear cells was counted in five randomly selected squares (103 x 103 microns). In 30 tissues, the number of mitochondria per cell was counted in five randomly selected cells from each lesion in transmission electron photomicrographs. Total cell counts in each group and number of chief cells showed no correlation with lesion detectability by scintigraphy. However, true positive lesions had a significantly higher number of oxyphil cells than false negative or normal glands. Twenty-one of 30 true-positive lesions had a oxyphil to-clear cell ratio > 1; in contrast to only two of 30 false-negative lesions and 0 of 30 normal glands (p < 0.0005). The number of mitochondria per cell was higher in oxyphil cells in true-positive lesions (adenomas: 155 +/- 58, hyperplasias: 55 +/- 18) than in chief or clear cells in false-negative or normal lesions (30 +/- 15, p < 0.001). Our data suggest that the detectability of abnormal parathyroid glands by 201TI/99mTc subtraction scintigraphy is in part dependent upon the presence of mitochondria-rich oxyphil cells. PMID- 8418266 TI - NDA sought for cancer imaging radiolabeled peptide. PMID- 8418267 TI - United States funds another study of nuclear blast test veterans...and declassifies data on bomb plant emissions. PMID- 8418268 TI - Technetium-99m-1,2-bis[bis(2-ethoxyethyl) phosphino]ethane: human biodistribution, dosimetry and safety of a new myocardial perfusion imaging agent. AB - A novel 99mTc complex (1,2-bis[bis(2-ethoxyethyl)phosphino] ethane, 99mTc tetrofosmin) has been developed to replace 201Tl in myocardial perfusion imaging. Biodistribution, safety and dosimetry of 99mTc-tetrofosmin were studied in 12 male volunteers, each at rest and during exercise. Safety parameters measured to 48 hr postinjection revealed no clinically significant long-term drug-related changes. Biodistribution was studied by acquiring whole-body or serial static images up to 48 hr postinjection. Technetium-99m-tetrofosmin shows good heart uptake (1.2%) with retention. Clearance is excellent from blood (< 5% by 10 min), liver (< 4.5% by 60 min) and lung. Sequestration of activity by skeletal muscle is enhanced during exercise. Radiation dosimetry calculations indicate that the effective dose, assuming a 3.5 hr bladder voiding period, is 32.9 x 10(-3) rad/mCi (8.9 x 10(-3) mSv/MBq) at rest and 26.7 x 10(-3) rad/mCi (7.1 x 10(-3) mSv/MBq) after exercise. Technetium-99m-tetrofosmin can produce high quality myocardial images from 5 min to several hours postinjection. PMID- 8418269 TI - Human pathologic correlation with PET in ischemic and nonischemic cardiomyopathy. AB - To assess the correlation between myocardial perfusion, metabolism and histologic findings in patients with cardiomyopathy, we evaluated myocardial perfusion and metabolism using positron emission tomography (PET) with 13NH3 (ammonia) and 18FDG (fluoro-2-deoxy-glucose) in nine patients prior to undergoing orthotopic cardiac transplantation. Four patients had ischemic cardiomyopathy (ISCM) and five had nonischemic cardiomyopathy (NISCM). Normalized circumferential profile analyses of representative mid-ventricular perfusion and metabolism PET images were performed for each patient. A corresponding mid-ventricular transaxial slice was obtained from the formalin fixed explanted heart and processed for routine histology. Hematoxylin-eosin stained and Masson trichrome stained sections were evaluated and the percentage of the slice occupied by infarct was determined planimetrically at 10-degree intervals in a circumferential manner. A significant correlation was found between circumferential normalized PET count density profile of perfusion and metabolism in both the ischemic and nonischemic groups (ISCM range r = 0.65-0.75; NISCM range, r = 0.70-0.87). Furthermore, there was a correlation in the ISCM group between the extent of matched perfusion/metabolism defects and transmural infarct involvement (r = 0.66-0.88). PET perfusion and metabolic data closely correlate with pathologic infarction in human hearts of ischemic cardiomyopathy patients. PMID- 8418270 TI - Renovascular hypertension: a perfusion disturbance that escaped recognition. AB - A bilateral, exercise-mediated, hippurate transport disturbance was previously described when patients with fixed renovascular hypertension were imaged with o iodo-hippurate. This study sought to test the hypothesis that patients with an abnormal exercise scintigram have a perfusion abnormality characterized by dysregulation of renal blood flow. We imaged 23 patients with hypertension and angiographically documented renovascular disease in the supine position, as well as during upright exercise. Seven normotensive volunteers served as controls. We measured the resting glomerular filtration rate (GFR) and the effective renal plasma flow (ERPF) with a single compartment radiotracer infusion clearance. The clearance examination also included a measurement period with 25 watt ergometric exercise. Nine hypertensive patients had normal exercise renograms. These patients had age-appropriate clearance values at rest and during exercise, as well as age-appropriate best-organ (generally without stenosis) GFR and ERPF values. The filtration fraction (FF) was 0.21 at rest and 0.22 during exercise. Fourteen hypertensive patients had a bilateral, exercise-induced disturbance of hippurate transport. In these patients, the global resting GFRs and ERPFs were decreased 40% from age-appropriate predicted values. The FF remained at 0.20. Light exercise caused a pronounced contraction of GFR and a less severe reduction in the ERPF. During exercise the mean filtration fraction was only 0.12. The exercise-induced reduction in the clearance values was bilateral, which indicated that the perfusion of nonstenosed organs was compromised as well. We suggest that the described perfusion abnormality occupies a relevant position during the maintenance phase of fixed renovascular hypertension. PMID- 8418272 TI - FDG-PET in oncology: there's more to it than looking at pictures. PMID- 8418271 TI - Myocardial uptake of metaiodobenzylguanidine in patients with left ventricular hypertrophy secondary to valvular aortic stenosis. AB - The time course of myocardial uptake of metaiodobenzylguanidine ([123I]MIBG) was studied in 26 patients: seven control subjects (Group 1) and 13 patients with left ventricular hypertrophy secondary to valvular aortic stenosis. Seven of these had received no treatment (Group 2) and six were receiving amiodarone or digoxin (Group 3); six heart transplant recipients were investigated for extra neuronal myocardial uptake of [123I]MIBG (Group 4). The index of myocardial [123I]MIBG uptake was lower in Groups 2 and 3 than in Group 1 (Group 2: 1.42 +/- 0.07, p < 0.001; Group 3: amiodarone, 1.30 +/- 0.10, p < 0.05; digoxin, 1.22 +/- 0.06, p < 0.01; Group I: 1.83 +/- 0.18) and lower in Group 3 than in Group 2. Patients of Group 4 showed a much lower mean index of myocardial [123I]MIBG uptake than the control group (1.07 +/- 0.08, p < 0.001). IN CONCLUSION: 1. Patients with left ventricular hypertrophy secondary to valvular aortic stenosis were found to have lower myocardial [123I]MIBG activity and rapid washout than the control subjects. 2. Amiodarone and digoxin partially inhibited myocardial [123I] MIBG uptake. 3. Extra neuronal myocardial uptake of [123I]MIBG in humans only accounts for 13% of the total cardiac activity. PMID- 8418273 TI - Colorectal cancer imaging with iodine-123-labeled CEA monoclonal antibody fragments. AB - This prospective, randomized multicenter study in 62 patients was designed to evaluate the efficacy and safety of radioimmunodetection (RAID) with 123I-labeled fragments, F(ab')2 and Fab', of IMMU-4, an anti-CEA monoclonal antibody (Immu RAID-CEA). It was found that ImmuRAID-CEA was safe and disclosed colorectal cancer sites at least 1 cm in size. The positive predictive value by lesions was 77% initially, and increased to 91% after 7 mo of follow-up. Only one patient developed a low level of HAMA. In 17 patients with 32 surgically confirmed lesions, there were 9% true-positive lesions for CT when RAID was false-negative, and 22% for RAID when CT was false-negative. Either CT or RAID detected all 32 lesions. In this small series, therefore, RAID was shown to complement CT findings by confirming suspected tumors and disclosing new lesions which had previously been occult. PMID- 8418274 TI - Quick diagnosis of hyperthyroidism with semiquantitative 30-minute technetium-99m methoxy-isobutyl-isonitrile thyroid uptake. AB - Ten normal volunteers and 14 patients with hyperthyroidism had pinhole thyroid imaging 30 min after an intravenous injection of 10 mCi 99mTc-MIBI. Technetium 99m-MIBI thyroid uptake was calculated by the formula: [total counts with a region of interest (ROI) over the whole thyroid gland] divided by [(mean counts of every pixel in the neck soft tissue) x (total number of pixels in ROI over the whole thyroid gland)]. The results showed that the 30 min 99mTc-MIBI thyroid uptake ratios had positive relationships with the 24 hr 131I-thyroid uptake (r = 0.79), and that the patients with hyperthyroidism had significantly higher 30 min 99mTc-MIBI thyroid uptake than the normal volunteers (5.31 +/- 0.78 s.e.m. versus 2.35 +/- 0.14 s.e.m., p < 0.005 using the Mann-Whitney U-test). Technetium-99m MIBI thyroid uptake may be useful for the rapid diagnosis of hyperthyroidism. PMID- 8418275 TI - Autoradiographic evaluation of the intra-tumoral distribution of 2-deoxy-D glucose and monoclonal antibodies in xenografts of human ovarian adenocarcinoma. AB - Monoclonal antibodies (Mabs) with tumor specificity and 2-[18F]-fluoro-2-deoxy-D glucose (FDG) are increasingly being used in cancer imaging. To better understand the mechanism and location of their uptake in tumors, nude mice bearing HTB77 IP3 ovarian cancer xenografts were injected intravenously with 3H-2-fluoro-2-deoxy-D glucose (3H-DG) or 14C-2-deoxy-D-glucose (14C-DG), with 125I-5G6.4, a Mab that was developed against the HTB77 IP3 cell line (specific Mab) or with 125I-UPC-10 (nonspecific Mab). Tumors were excised at 2 hr or 6 days postinjection and autoradiography was performed. In tumors from animals receiving the Mabs, the grains were concentrated mainly on tumor cells near connective tissue ridges bearing blood vessels at 2 hr postinjection. At 6 days postinjection, further penetration into the tumors was seen, but four times more grains were seen on the viable cancer cells in the tumors receiving the specific Mab than in those receiving the irrelevant Mab. The selective accumulation of 2-DG in viable cancer cells and the negligible concentration of 2-DG in necrotic areas may be preferable to tumor-specific Mabs in assessing the extent of viable tumor and treatment response. Knowledge of the localization of a tracer at the microscopic level is essential to the understanding of the signal depicted on nuclear images using either type of radiotracer. PMID- 8418276 TI - Validation of nitrogen-13-ammonia tracer kinetic model for quantification of myocardial blood flow using PET. AB - Positron emission tomography has been shown to provide quantitative estimates of myocardial blood flow using 13N-ammonia and 15O-water. In a validation study, myocardial blood flow was noninvasively determined in 11 open-chest anesthetized dogs using dynamic positron emission tomography. The radiopharmaceuticals 13N ammonia and 15O-water were intravenously administered and measurements were carried out at rest and following pharmacological vasodilation to assess blood flow over a range from 53 to 580 ml/100 g/min. Quantification of blood flow based on tracer kinetic modeling of 13N-ammonia data correlated closely with myocardial blood flow determined by microspheres (y = 0.944 x +7.22, r = 0.986) and with the 15O-water injection technique y = 1.054 x -15.8 (r = 0.99). The use of 13N ammonia with positron emission tomography enables the accurate quantification of myocardial blood flow. Using this technique, uncomplicated study protocols simplify the measurement procedures while providing excellent qualitative and quantitative information. PMID- 8418277 TI - Joint uptake and body distribution of a technetium-99m-labeled anti-rat-CD4 monoclonal antibody in rat adjuvant arthritis. AB - Joint uptake and body distribution of a 99mTc-labeled monoclonal antibody (Mab) to the rat CD4 molecule (W3/25; IgG1) were investigated after intravenous injection in normal rats and in animals with experimentally induced adjuvant arthritis. An isotype-matched Mab with irrelevant specificity (anti-human carcino embryonic-antigen) was used as control. A 4 hr sequential gamma-camera imaging revealed that both anti-CD4 and control Mab accumulated to a higher degree in arthritic than in normal ankle joints; the accumulation was comparable for the two Mabs. In contrast to the inflamed joints, a specific accumulation of the anti CD4 Mab was found in organs rich in CD4-positive cells, i.e. spleen, bone marrow and lymph nodes, as assessed by direct well counter measurements 16 hr after injection. The control Mab displayed no preferential organ accumulation in either normal or diseased animals. These results indicate that a specific accumulation of anti-CD4 Mabs occurs in CD4-positive-cell-rich tissues in both normal and diseased animals and that immunoglobulins accumulate preferentially in inflamed joints regardless of their antibody specificity. PMID- 8418278 TI - Brain uptake of thallium-201 from the cerebrospinal fluid compartment. AB - The present report elucidates the movement of 201Tl through the cerebrospinal fluid compartment and its subsequent uptake by normal brain. Autoradiographic studies of rat brain, after stereotaxic 201Tl injection into either the lateral or fourth ventricle, reveal that 201Tl moves freely through the cerebrospinal and extracellular fluid compartments. Subsequent to lateral ventricular injection, central thalamic and specific hypothalamic nuclei are heavily labeled. Densely labeled mesencephalic nuclei include the periaqueductal grey and oculomotor nuclear complex. Labeling of giant cells within the vestibular complex is suggestive of neuronal uptake. Major fiber tracts are devoid of label confirming that 201Tl uptake does not occur in white matter. Most labeling following fourth ventricle injection occurs within the caudal medulla and cervical spinal grey. Our findings suggest that 201Tl uptake by normal brain from the cerebrospinal fluid occurs as a function of thallium concentration and neuronal activity. PMID- 8418279 TI - Basics for beginning a parish nurse program. PMID- 8418280 TI - How do parish nurses help people? A research perspective. PMID- 8418281 TI - Mercy model: church-based health care in the inner city. PMID- 8418282 TI - Helping children deal with death. PMID- 8418283 TI - The power of the pen. PMID- 8418284 TI - Reclaiming the Church's healing role. PMID- 8418285 TI - Sharing the burden. PMID- 8418286 TI - Nurses in the church: profiles of caring. PMID- 8418287 TI - The intestine and human immunodeficiency virus. PMID- 8418288 TI - Serogroups, serotypes and subtypes of Neisseria meningitidis isolated from patients and carriers in Greece. AB - The increase in the number of cases of meningococcal disease reported to the Ministry of Health in Athens since 1989 prompted the present study to determine if isolates from patients or carriers expressed the same phenotypic characters as those in other parts of Europe. None of the isolates from patients (31) or carriers (547) expressed the antigenic combinations associated with outbreaks in northern Europe, i.e., B:15:P1.16 or B:4:P1.15. The majority of the Greek isolates did not react with any of the six monoclonal serotype reagents tested; however, most reacted with one or more of the 11 monoclonal subtype antibodies. The results suggest that additional serotype reagents are needed for epidemiological studies in southeastern Europe and that vaccines based on serotype antigens developed against outbreak strains in northern Europe would not be effective in Greece. PMID- 8418289 TI - Molecular epidemiology of chronic pulmonary colonisation by Pseudomonas aeruginosa in cystic fibrosis. AB - The epidemiology of pulmonary colonisation by Pseudomonas aeruginosa was studied in 21 patients with cystic fibrosis (CF) by field inversion gel electrophoresis. DraI-DNA restriction patterns were analysed for 187 P. aeruginosa isolates from these patients. The results revealed that the strains present in individual patients varied during the course of chronic colonisation; the emergence of new strains often was associated with periods of antibiotic therapy. Patients often were colonised by more than one strain (two or three strains were present in 54% of the patients) and the strains obtained from unrelated patients were highly heterogeneous, in contrast to those isolated from a pair of twins. These results demonstrate the heterogeneity and variability of P. aeruginosa isolates in the pulmonary flora of chronically infected CF patients. PMID- 8418290 TI - Lior-serotype variants in Campylobacter isolates from the same stool sample. AB - Campylobacter strains isolated from the same stool sample were characterised by determination of biochemical properties and both heat-labile (Lior) and heat stable (Lauwers) serotypes. In six of 60 campylobacter-infected stools, two or three strains differing in Lior-serotype were isolated from the same stool. In four of these six cases, the isolates with different Lior-serotypes showed identical biochemical reactions and identical heat-stable antigenic patterns. A predominant Lior-serotype was not detected among them but Lauwers-antigens O:3, O:14 and O:16 were found in isolates from three of the six stool samples. Moreover, the isolates were identified as C. coli in 76.5% of the stool samples (p < 0.05). We believe that variation in heat-labile antigens occurs in vivo and might be associated particularly with certain heat-stable serotypes of C. coli. PMID- 8418291 TI - Measurement of Toxoplasma IgM by a microparticle capture enzyme immuno-assay. AB - The microparticle capture enzyme immuno-assay (MEIA) is an automated system for measuring specific antibody by interaction with antigen-coated particles. An MEIA method for detecting toxoplasma-specific IgM was compared with established reference methods. The MEIA had an acceptable level of sensitivity and reproducibility and was easier to perform than conventional tests, but it required expensive, dedicated equipment and, in our study, false positive results were recorded with 7% of samples. MEIA could be used to investigate immunocompetent patients with suspected toxoplasmosis but positive findings should be confirmed by an alternative form of assay. The technique is not suitable for the investigation of neonates, for which a more sensitive method is required. PMID- 8418292 TI - Toxoplasma polymerase chain reaction on experimental blood samples. AB - A two-stage polymerase chain reaction (PCR) assay employing oligonucleotide primers from the B1 gene of Toxoplasma gondii was developed and assessed for sensitivity and specificity. It was able to detect T. gondii DNA from as little as one parasite/sample in mock-infected rat or mouse leucocyte preparations. Parasitaemia was also identified in animals at five stages between 16 and 66 h after infection with the virulent RH strain, and at 12 stages between 2 and 38 days after infection with the cyst-forming Beverley strain. In the latter case, PCR was more sensitive than animal culture. No cross-reactions were observed in samples containing various opportunist pathogens which may also be found in the blood of immunocompromised patients. PMID- 8418293 TI - Haemagglutinating activity, serum sensitivity and enterotoxigenicity of Aeromonas spp. AB - Of 97 isolates of Aeromonas spp. that were examined for haemagglutination (HA) and enterotoxigenicity, 35 were from clinical and 62 from environmental sources; 66 of them were also screened for sensitivity to normal human serum (NHS). HA was caused by 44 isolates (45%); it was unrelated to the source of the strain, but it was caused by a higher proportion of the isolates of A. hydrophila than of A. sobria or A. caviae. Of the haemagglutinating strains, 82% were enterotoxigenic, whereas most of the non-haemagglutinating strains were non-toxigenic when tested initially. All the latter became enterotoxin producers after serial passage through rabbit ileal loops, but without change in HA. Most (64%) of the isolates, including 68% of A. caviae (72% of clinical and 65% of environmental), were resistant to the bactericidal action of NHS. Most (92%) of the serum-sensitive strains were killed by activation of both the classical and alternate pathways of complement, the others only by the alternate pathway. Most (74%) of the serum resistant strains caused fluid accumulation in the initial tests in ileal loops, regardless of species or source. Haemagglutinating and serum-resistant strains caused significantly more accumulation of fluid (p < 0.05) than non haemagglutinating and serum-sensitive strains. This study shows partial correlation between HA or serum sensitivity and enterotoxigenicity, but the properties are probably not genetically linked. PMID- 8418294 TI - Effects of selected surfactants on purified glucosyltransferases from mutans streptococci and cellular adherence to smooth surfaces. AB - The inhibitory effect of non-ionic, anionic, cationic and ampholytic surfactants on cellular growth of Streptococcus mutans MT8148 and S. sobrinus 6715, on glucan synthesis by the purified glucosyltransferase (GTase) from these organisms, and on bacterial adherence to glass surfaces was examined in vitro. Cationic surfactants exhibited marked bactericidal activities. Anionic and ampholytic compounds were less strongly bactericidal and non-ionic surfactants produced only slight inhibition of cell growth under the conditions tested. Some non-ionic compounds had no effect on this. Glucan synthesis by GTase from mutans streptococci was inhibited by anionic and cationic surfactants. Among various GTase proteins, insoluble glucan synthesising GTases, i.e., S. mutans CA-GTase and S. sobrinus GTase-I were those most effectively inhibited by these agents. However, it was noted that whereas lower concentrations of cationic surfactants enhanced these GTase activities, higher concentrations of the surfactants were inhibitory. Non-ionic detergents stimulated soluble glucan synthesis from S. mutans CF-GTase and cationic and ampholytic surfactants enhanced or inhibited glucan synthesis depending on the concentrations of the surfactants. Sucrose dependent cellular adherence of resting cells of mutans streptococci to glass surfaces was inhibited by the addition of surfactants that annulled the GTase activities. PMID- 8418295 TI - Biotype and molecular fingerprints of metronidazole-resistant strains of Helicobacter pylori from antral gastric mucosa. AB - Biotypes, ribosomal RNA gene restriction patterns (ribopatterns), whole-cell protein patterns and plasmid profiles of paired Helicobacter pylori isolates from 17 patients were examined. Each pair comprised a pre- and a post-treatment isolate; nine of the 17 post-treatment isolates were obtained after treatment with tripotassium dicitrate bismuthate (De-Nol) and metronidazole. All strains of H. pylori had identical biotypes, but exhibited diversity between pairs in their molecular fingerprints. Each of the 17 strain pairs had unique ribopatterns; the pre- and post-treatment isolates in most pairs (16 of 17) were similar or identical, irrespective of metronidazole susceptibility. DNA subtype variants were detected in three patient sets. Although nine post-treatment isolates had acquired resistance to metronidazole, most (six of nine) resembled the pre treatment isolates in their ribopattern, protein and plasmid profiles. No significant correlation was observed between metronidazole resistance and plasmid content in these H. pylori isolates. Emergence of post-treatment metronidazole resistant isolates of H. pylori was associated only rarely with colonisation by a novel strain or acquisition of a plasmid and, in most patients, probably resulted from spontaneous emergence of resistance in the original infecting strain. PMID- 8418296 TI - Survival of group A streptococci in dried human blood. AB - The resurgence of streptococcal infections in the USA and Europe and their high incidence in other parts of the world prompted an examination of the survival and maintenance of virulence of group A streptococci. Human blood containing group A streptococci was placed on small pieces of sterile paper towelling and allowed to dry at room temperature. At periods of 2, 8, 15 and 20 weeks later, the paper with the dried blood was placed in Todd-Hewitt broth and incubated at 37 degrees C overnight. All the samples tested at 2 weeks grew in broth, and with only one exception, grew in fresh human blood provided by five donors. At 8 weeks only two of the 10 strains failed to grow in broth; seven of the eight viable cultures also grew in blood. At 15 and 20 weeks after drying the eight cultures were still viable. Since seven were able to grow in fresh blood as well as in broth it is assumed that their virulence factor(s) had been retained. PMID- 8418297 TI - An electronmicroscope study of the effect of sulphadiazine and trimethoprim on Enterobacter cloacae. AB - Electronmicroscopy of thin sections of log phase cells of Enterobacter cloacae NCTC 10005 grown for 4 h in the presence of sulphadiazine 250 micrograms/ml, trimethoprim 12.5 microliters/ml or the combination of sulphadiazine 250 micrograms/ml plus trimethoprim 12.5 micrograms/ml indicated that both agents caused marked morphological damage even though the MIC of sulphadiazine for the E. cloacae strain was > 3000 micrograms/ml. The damage took the form of electron transparent areas devoid of ribosomes in the cytoplasm and detachment of the outer membrane. The latter was most marked with trimethoprim, which also caused damage to the cytoplasmic membrane. It is postulated that the synthesis of the peptidoglycan layer was affected by the antimetabolites since the morphological effects were strikingly similar to those caused by treatment of E. cloacae with disodium edetate plus lysozyme. Viable counts of cultures undergoing the same treatments as those prepared for electronmicroscopy indicated that although sulphadiazine merely partially inhibited growth it nevertheless enhanced the bactericidal action of trimethoprim over a 5-h period. PMID- 8418298 TI - Purification and characterisation of intracellular toxin A of Clostridium difficile. AB - After sonic disintegration of Clostridium difficile cells, intracellular toxin A was purified to homogeneity by thyroglobulin affinity chromatography (TGAC) followed by anion-exchange (Mono Q) by fast protein liquid chromatography (FPLC). High haemagglutinating (HA) activity was detected in TGAC-unbound fractions (2(9)/50 microliters), but not in TGAC thermal eluates (2(0)/50 microliters). The low HA titre of the thermal eluates was markedly increased to 2(5)/50 microliters after dialysis against 0.02 M Tris-HCl (pH 7.5). A disparity in the position of the peaks containing cytotoxic and HA activity was observed in the first Mono Q FPLC step. Intracellular toxin A without HA activity was obtained by a second Mono Q-FPLC step. The M(r) of the intracellular toxin A was estimated by polyacrylamide gel electrophoresis (PAGE) to be 580 kDa under non-denaturing conditions. The minimum doses of the toxin causing cytotoxicity, mouse lethality and enterotoxicity were 0.83 ng, 8.7 ng and 5 micrograms, respectively. PMID- 8418299 TI - Proceedings of the Pathological Society of Great Britain and Ireland. 166th meeting. London, 6-8 January 1993. Abstracts. PMID- 8418300 TI - Increasing mammography utilization: a controlled study. AB - BACKGROUND: Despite the effectiveness of breast cancer screening for women older than 50 years of age, only about one third of these women in the United States receive annual mammography. PURPOSE: This study was designed to determine if a community-wide intervention could increase use of mammography screening for breast cancer. Secondary end points were determination of changes in women's knowledge and attitudes toward mammography and physicians' self-reported screening practices. METHODS: We conducted a controlled study from January 1987 through January 1990 in two eastern North Carolina communities--New Hanover County (the experimental community) and Pitt County (the control community). Before development and implementation of the intervention program in New Hanover County and after the program had been in operation for 1 year, 500 women of ages 50-74 years and all primary-care physicians in each community were interviewed by telephone. In these interviews, we determined the use of mammography for breast cancer screening and the knowledge and attitudes about it. We also established the number of screening mammograms performed in 1987 and 1989 in each county and reviewed medical records to determine the percentage of women the physicians had referred for mammograms. RESULTS: The percentage of women who reported receiving a mammogram in the previous year increased from 35% to 55% in the experimental community and from 30% to 40% in the control community (difference of differences, 10%; P = .03 after adjustment for race, education, age, and having a regular doctor; 95% confidence interval, 1%-18%). Increases were greater in New Hanover County regardless of age, race, income, and education. However, the increase was less for Black women than for White women, both overall and in most demographic subgroups. The total number of mammograms performed increased 89% in the experimental community and 45% in the control community. Women's knowledge about mammography changed little, but the intention to get a mammogram increased 30% in New Hanover County, compared with a 17% increase in Pitt County--a statistically significant difference (P < .01). Physician reports and medical record reviews in the two communities showed similar increases in the number of mammograms ordered. CONCLUSIONS: A community-wide effort to increase use of breast cancer screening was successful, but more work must be done to reach the National Cancer Institute's goal of annual mammograms for 80% of women of ages 50 74. PMID- 8418301 TI - Effects of modulation of basic fibroblast growth factor on tumor growth in vivo. AB - BACKGROUND: Recombinant human basic fibroblast growth factor (rHu-bFGF) is known to stimulate proliferation in some tumor cells and to modulate tumor vascularization. PURPOSE: The purpose of this study was to examine the possible role of this agent in the development of tumors. The study was designed to determine the effects of modulating bFGF activity in vivo in tumor models from cell lines with different responses to bFGF and with different content and receptor levels of bFGF. METHODS: Two tumor cell lines (human DLD-2 colon carcinoma and rat C6 glioma) were characterized for bFGF content and bFGF receptor levels by Western blot analysis in cultured cells and by studies of [125I]rHu-bFGF binding to sections from xenografts grown in nude mice. Tumor cell proliferation was monitored after treatment with rHu-bFGF or the DG2 or DE6 IgG monoclonal antibody to rHu-bFGF in culture and in vivo. RESULTS: C6 cells exhibited 7800 high-affinity receptors for rHu-bFGF per cell (dissociation constant [Kd] = 46 pM), while DLD-2 cells lacked high-affinity receptors. rHu bFGF stimulated [3H]thymidine uptake by C6 cells, but the addition of DG2 IgG prevented this stimulation; rHu-bFGF had no effect on [3H]thymidine incorporation by DLD-2 cells. C6 cells had higher levels of immunoreactive bFGF than did DLD-2 cells. The xenografts from both cell lines exhibited high-affinity [125I]rHu-bFGF binding that was concentrated on vascular-like structures. rHu-bFGF at a dosage of 0.25 mg/kg given intraperitoneally daily for 18 days caused a twofold increase in DLD-2 tumor weight but had little effect on the growth of C6 xenografts. In contrast, daily intravenous injections of DG2 IgG given to mice had no effect on DLD-2 tumor growth but reduced growth of C6 tumors by approximately 30%--a statistically significant difference. CONCLUSIONS: The addition of exogenous rHu bFGF or of a neutralizing antibody resulted in significant alterations in tumor growth in vivo, which were specific for tumor type and bFGF characteristics. While some of these effects may be mediated by the bFGF-responsive endothelial cells of the tumor vasculature (DLD-2 colon carcinoma), others may result from inhibition of bFGF-dependent tumor cell proliferation (C6 glioma). IMPLICATIONS: Studies that measure tumor blood flow are necessary to confirm that these effects are mediated by changes in tumor vasculature. PMID- 8418303 TI - Characteristics relating to ovarian cancer risk: collaborative analysis of seven U.S. case-control studies. Epithelial ovarian cancer in black women. Collaborative Ovarian Cancer Group. AB - BACKGROUND: Previous epidemiologic studies of ovarian cancer have focused chiefly on White women, who have a higher incidence of ovarian cancer than Black women. No study has previously examined risk factors for ovarian cancer among Black women. PURPOSE: This study was designed to evaluate the risk of epithelial ovarian cancer in Black women in relation to reproductive characteristics such as pregnancy, oral contraceptive use, and breast-feeding, and to determine whether differences in reproductive factors between Black and White women account for differences in ovarian cancer incidence. METHODS: Combining interview data from seven case-control studies, we compared reproductive characteristics of 110 Black case subjects with a diagnosis of epithelial ovarian cancer between 1971 and 1986 with characteristics of 251 Black population control subjects and 114 Black hospital control subjects. We also compared the prevalence of reproductive factors in 246 Black population control subjects and 4378 White population control subjects and estimated the fraction of Black-White differences in ovarian cancer incidence attributable to racial differences in prevalence of these characteristics. RESULTS: Decreased risks of epithelial ovarian cancer in Black women were associated with parity of four or higher (odds ratio [OR] = 0.53; 95% confidence interval [CI] = 0.25-1.1), breast-feeding for 6 months or longer (OR = 0.85; 95% CI = 0.36-2.0), and use of oral contraceptives for 6 years or longer (OR = 0.62; 95% CI = 0.24-1.6). A greater proportion of Black women (48%) than White women (27%) reported four or more term pregnancies, and Black women (62%) were more likely than White women (53%) to have breast-fed their children. Oral contraceptive use was more common among White women (59%) than Black women (51%). CONCLUSION: Differences in the prevalence of other factors related to ovarian cancer risk or differences in genetic susceptibility must explain most of the Black-White differences in incidence of ovarian cancer. PMID- 8418302 TI - Calcium and colorectal epithelial cell proliferation: a preliminary randomized, double-blinded, placebo-controlled clinical trial. AB - BACKGROUND: Colonic epithelial cell proliferation is increased in patients at high risk for colon cancer. Calcium administration has ameliorated the proliferative changes in rodents, and findings in small, uncontrolled clinical trials have suggested similar effects in humans. PURPOSE: This preliminary, double-blind, randomized clinical trial was designed 1) to investigate whether supplemental calcium will reduce colonic epithelial cell proliferation in patients with sporadic adenomas who consume a high-fat, Western-style diet; 2) to determine the sample size (number of scorable crypts per person) needed to achieve adequate statistical power; and 3) to evaluate the feasibility of full scale clinical trials. METHODS: Twenty-one sporadic adenoma patients were treated daily with placebo or 1200 mg of supplemental calcium. To determine colonic epithelial cell proliferation, we used tritiated thymidine labeling of colon crypt epithelial cells in rectal biopsy specimens and calculated the percentage of labeled cells (labeling index [LI]). Two pathology technician "readers" independently scored each specimen, and inter-reader reliability was determined. Subjects remained on their usual diet during the study, and intake of calories, calcium, total fat, and vitamin D did not differ substantially among them. We calculated curves for statistical power to determine the number of scorable crypts needed per person for detection of a statistically significant difference (P < .05) of 1.0% in mean LI. RESULTS: The pooled baseline LI was 4.7%. In the calcium-treated group, the LI increased 0.6% (proportional increase, 12.8%); in the placebo-treated group, it decreased 0.5% (proportional decrease, 10.6%). The difference between change in the mean LI from baseline to 8 weeks' follow-up in the placebo group versus the calcium group was not statistically significant. The intraclass correlation coefficient for inter-reader reliability for the baseline LI was .66. Analyses indicated scoring eight crypts sufficient for estimates of the LI adequate for between-group comparisons, a level achieved in 81% of biopsy specimens. CONCLUSIONS: Calcium carbonate supplements delivering 1200 mg elemental calcium daily may not decrease colonic epithelial cell proliferation over an 8-week period in sporadic adenoma patients. In future trials measuring the LI, consideration should be given to ensuring adequate numbers of scorable crypts and to the impact of inadequate biopsy procedures, labeling failure, reader reliability, and participant withdrawal. Our findings support the feasibility of a full-scale clinical trial to further study the relationships among dietary calcium, colonic epithelial cell proliferation, and colorectal cancer. PMID- 8418304 TI - Increased nm23-H1 and nm23-H2 messenger RNA expression and absence of mutations in colon carcinomas of low and high metastatic potential. AB - BACKGROUND: The murine nm23 gene suppresses the metastatic behavior of malignant rodent tumor lines, and reduced nm23 expression correlates with increased likelihood of lymph node metastases in human breast cancers. More recent data have demonstrated the existence of two human nm23 gene homologues, nm23-H1 and nm23-H2, and have shown that deletion of nm23-H1 alleles occurs in some colon carcinomas associated with poor prognosis. These findings suggest that nm23-H1 encodes for suppression of colon carcinoma metastasis. In contrast, we have previously reported that total nm23 messenger RNA (mRNA) expression is increased to similar levels in colon tumors of both high and low metastatic potential. PURPOSE: This study was designed to reconcile our previous findings with the recent report of nm23-H1 allelic deletion in human colon cancers associated with poor prognosis. Our purpose was to examine human colon cancers for inactivation of two candidate metastasis suppressor genes, nm23-H1 and nm23-H2, either by mutation or by loss of gene transcription. METHODS: We used ribonuclease protection assays to analyze human colon tumors for the level of nm23-H1 (43 samples) and nm23-H2 (41 samples) transcript (mRNA) expression and the presence of mutations that could inactivate potential suppressor function. RESULTS: We detected only wild-type nm23-H1 and nm23-H2 mRNA. Expression of nm23-H1 mRNA increased in 33 of 41 colon tumors, and expression of nm23-H2 mRNA was elevated in 28 of 41 colon tumors relative to that in matched normal mucosa. Increases in these mRNA levels were similar in tumors of both low and high metastatic potential. CONCLUSIONS: These results suggest that, despite correlation of nm23 H1 allelic deletions with colon cancers associated with poor prognosis, nm23-H1 and nm23-H2 alleles do not directly mediate metastasis suppression in colon carcinoma. Our results leave unexplained the observation that nm23-H1 allelic deletion correlates with metastatic potential of colon carcinomas. IMPLICATIONS: These findings also contrast with the demonstration of nm23 metastasis suppressor activity in murine melanoma and with the correlation of loss of nm23 expression in breast cancer with poor prognosis. It may be that metastasis suppression by the nm23 gene is a tissue-specific phenomenon. PMID- 8418305 TI - Expression of the retinoblastoma susceptibility gene in childhood rhabdomyosarcomas. AB - BACKGROUND: Alveolar and embryonal rhabdomyosarcomas are soft-tissue tumors that occur mainly in childhood. The more aggressive alveolar subtype has been found to possess a characteristic chromosomal abnormality located near the retinoblastoma susceptibility gene (RB1). RB1 is a tumor suppressor gene implicated in the development of retinoblastoma and other, unrelated malignancies, especially osteogenic sarcomas and other second malignancies in retinoblastoma survivors. PURPOSE: The goals of our study were (a) to determine whether abnormalities of RB1 occur routinely in sporadic rhabdomyosarcomas, as reported for other sporadic malignancies, especially bone and soft-tissue sarcomas of adulthood; and (b) to assess differences in the functional status of the gene in embryonal and alveolar rhabdomyosarcomas that might explain the differing clinical aggressiveness of these two variants. METHODS: Analyses of messenger RNA (mRNA) and protein expression and immunohistochemical studies were performed on 11 rhabdomyosarcoma cell lines from patients with no family history of retinoblastoma, and RB1 protein expression was studied by immunoprecipitation in primary tumor biopsy tissue from 18 patients with sporadic rhabdomyosarcoma (five embryonal and 13 alveolar). RESULTS: The RB1 gene appears to be normal in structure, expression, and function and is comparably phosphorylated in both forms of childhood rhabdomyosarcoma. Normal-size RB1 mRNA was present in all rhabdomyosarcomas, whereas clearly abnormal expression was documented in the controls, as expected: mRNA transcripts were truncated in Y79 retinoblastoma and absent in DU4475 breast carcinoma. In addition, immunoprecipitation with antibody to RB1 protein indicated the presence of normal RB1 protein in all rhabdomyosarcomas. CONCLUSIONS: These findings are distinct from those for other adult asarcomas, in which RB1 expression frequently is reported as abnormal. They are particularly surprising in view of the high incidence of RB1 abnormalities in osteosarcoma, the bone tumor most associated with retinoblastoma. The etiology and biologic behavior of rhabdomyosarcoma are, thus, unlikely to be dependent on RB1 mutations. IMPLICATIONS: The findings reported here clearly imply that the RB1 gene is structurally and functionally normal in childhood rhabdomyosarcoma. Other, as yet unidentified, genetic defects are apparently etiologic in this particular sarcoma. PMID- 8418306 TI - Ovarian cancer among black women. PMID- 8418307 TI - Fertility drugs may raise ovarian cancer risk. PMID- 8418308 TI - Cancer patient groups grow in democratic Hungary. PMID- 8418309 TI - New technologies proving useful for breast cancer diagnosis. PMID- 8418310 TI - Opportunities "opening up" for gene therapy. PMID- 8418311 TI - Moving forward with a "second generation" lymphoma vaccine. PMID- 8418312 TI - Myths about cancer lead to workplace discrimination. PMID- 8418313 TI - Overview of combined modality therapies for head and neck cancer. AB - The survival rate is 40% for patients with advanced squamous cell carcinoma of the head and neck whose tumors are completely resected and 20% for those with unresectable tumors treated with radiotherapy alone. During the past 10 years, combined modality approaches have been developed in an effort to enhance locoregional disease control, reduce distant metastases, and preserve anatomic function. They include the following: (a) neoadjuvant chemotherapy followed by standard therapy with surgery and/or radiation, (b) adjuvant chemotherapy after surgery or radiotherapy with or without neoadjuvant chemotherapy, and (c) neoadjuvant chemotherapy concurrent with radiotherapy. Even early studies of cisplatin plus fluorouracil (5-FU) reported 50%-90% overall response rates, and this is the main drug combination used in clinical trials. In the Veterans Affairs Cooperative Study Program, 5-FU and cisplatin followed by radiotherapy achieved a 49% complete response rate and preservation of the larynx in 64% of the patients. These results supported the findings of other nonrandomized trials that sequential induction chemotherapy and radiotherapy results in laryngeal preservation without compromising overall survival. The Head and Neck Cancer Intergroup Trial compared adjuvant postoperative cisplatin plus 5-FU prior to radiotherapy with postoperative radiotherapy. Survival at 4 years was 44% with radiotherapy alone and 48% with chemotherapy and radiotherapy. Biochemical modulation of 5-FU with leucovorin and biologic response modifiers such as interferon has achieved complete response rates as high as 66%, but severe mucositis continues to be the dose-limiting toxic effect. Standard radiotherapy for advanced nasopharyngeal carcinoma--a unique type of head and neck cancer- resulted in 5-year survival of 10%-40%, but neoadjuvant chemotherapy plus radiotherapy has achieved overall complete response rates greater than 80% with median survival of 5 or more years. We conclude that curability of nasopharyngeal carcinoma with a combined modality approach appears to be an achievable goal, but adequate evaluation in large-scale randomized trials is hampered by low accrual to clinical trials. In summary, neoadjuvant therapy for squamous cell head and neck carcinoma results in complete response rates of 22%-66%, but addition of adjuvant therapy may be necessary for a survival advantage over standard therapy. Although concurrent chemoradiotherapy has produced increased survival, additional trials are needed to determine optimal dosages. PMID- 8418314 TI - Thoracic magnetic resonance imaging: physics and pulse sequences. AB - The article summarizes the basic concepts of magnetic resonance (MR) physics so that radiologists can optimize the images obtained in MR imaging of the chest. The specification of thoracic MR scan protocols is more complex than for other body regions because of cardiac and respiratory motion and blood flow. Various motion suppressing techniques such as motion compensation, respiratory compensation, and spatial presaturation have been developed, but their appropriate application is facilitated by a thorough understanding of the imaging process. Magnetic susceptibility and low proton density limit the usefulness of MR imaging in the assessment of the lung parenchyma. Nevertheless, recent studies have shown considerable improvement in image quality with the use of short echo times. Suggested imaging protocols for the evaluation of chest wall, mediastinum, hilum, heart, and great vessels are discussed. PMID- 8418315 TI - Magnetic resonance imaging of lung parenchyma. AB - Imaging of the lungs presents a unique challenge for MR technology. Signal from lung parenchyma is limited by low proton density, the effects of motion and susceptibility, and a multiexponential T2 relaxation time. We have illustrated the use of spin echo and projection reconstruction MR sequences in imaging lung parenchyma, and the use of a GRE sequence in imaging the pulmonary vascular tree. PMID- 8418316 TI - Clinical utility of Gd-DTPA-enhanced magnetic resonance imaging in lung cancer. AB - Magnetic resonance (MR) imaging has both advantages and disadvantages in its application in lung cancer staging. Because of its ability to provide superior contrast resolution and to display structures in many planes, MR imaging is better than computed tomography (CT) for the detection of mediastinal and chest wall invasion. MR imaging also is more sensitive than CT for detection of hilar and mediastinal lymph node enlargement. Multiplanar T1- and T2-weighted images are optimal for differentiating lymph nodes from large vessels without the need for contrast enhancement; in these cases administration of Gd-DTPA provides no more information than plain MR images. MR studies should be used for examining patients with suspected mediastinal or chest wall invasion and those who have equivocal hilar or mediastinal adenopathy. The shortening effect of Gd-DTPA on the T1 value results in a high signal. This effect is dependent upon both the perfusion and diffusion of the contrast agent and the amount of extracellular fluid. The distribution of Gd-DTPA is similar to that of iodinated water-soluble contrast media. Gd-DTPA examination should be tailored to provide information regarding blood flow, vascularity, and permeability, none of which is easily appreciated on CT or plain MR images. Applications for which Gd-DTPA enhancement may be helpful include differentiating between malignant and benign pulmonary masses, differentiating between hilar lung cancer and peripheral postobstructive atlectasis or pneumonia, determining therapeutic effect after radiation therapy, and differentiating between recurrent or residual tumor and radiation pneumonitis. PMID- 8418317 TI - Magnetic resonance imaging of the mediastinum. AB - Magnetic resonance (MR) imaging is becoming a primary modality for evaluating the mediastinum. MR affords multiplanar imaging capabilities without exposing patients to ionizing radiation. The inherent contrast effect of different mediastinal tissues sharply delineates anatomic structures on MR images without contrast enhancement. Gradient-echo and phase-mapping techniques permit noninvasive qualitative and quantitative assessment of mediastinal blood flow. High soft tissue contrast and flow analysis capabilities make MR imaging a valuable modality for evaluating mediastinal vascular disorders. Various mediastinal tumors and their extent are best identified by the use of T1 weighted, T2-weighted, and gadolinium-enhanced images. Both primary and secondary chest wall lesions may be assessed with standard spin-echo MR images. Complex pleuroparenchymal lesions may be evaluated by means of a multiplanar approach and modified pulse sequences. This article addresses the technical parameters governing MR imaging of the mediastinum and describes MR characteristics of various pathologic conditions. PMID- 8418318 TI - Interstitial lung disease in rheumatoid arthritis: assessment with high resolution computed tomography. AB - Interstitial lung disease (ILD) is a frequent manifestation of rheumatoid arthritis (RA), and it has a close bearing on the prognosis of RA patients. Computed tomography (CT) has been shown to be excellent for the diagnosis of diffuse lung disease. In this study chest radiographs and high-resolution CT (HRCT) scans were obtained in 91 patients with RA to evaluate their ILD precisely. By HRCT 43 patients could be diagnosed as having interstitial pneumonitis (IP), and 5 could be diagnosed as having bronchiolitis. The remaining 43 patients were normal by HRCT. Chest radiographic findings were consistent with the HRCT findings in approximately 50% of patients with IP. HRCT was superior to chest radiographs for the detection of early interstitial changes. The histogram of HRCT values might be a useful adjunct to HRCT diagnosis by adding some degree of objectivity. HRCT is useful for the diagnosis of ILD in patients with RA. PMID- 8418319 TI - Multiple primary lung cancers: radiographic and bronchoscopic diagnosis. AB - The article reports a review of 1140 cases of primary lung cancer diagnosed between 1974 and 1987 (13-year period) to examine the clinicopathologic features of multiple primary lung cancer. The following criteria were used: There were no malignant tumors in other organs; the tumors were separate or located in different lobes; the tumors were of different histologic types; if the histologic types were similar, there was no carcinoma in the common lymphatics of any tumors and no extrapulmonary metastases at the time of diagnosis. The following results were obtained. The incidence was 1.2% (14 of 1140 cases); 11 patients were heavy smokers; the most common histologic type was squamous cell carcinoma, and the combination of two squamous cell carcinomas was most common; 12 cases were bilateral, and 11 cases were synchronous; 19 tumors were diagnosed by bronchoscopic biopsy; 9 patients with bilateral tumors received chemotherapy and radiation therapy. PMID- 8418320 TI - Hypersensitivity pneumonitis: a diagnostic dilemma. AB - Hypersensitivity pneumonitis is an important but underdiagnosed form of interstitial lung disease. Its clinical hallmark is recurrent or chronic pulmonary symptoms of variable severity associated with an array of systemic symptoms but without extrapulmonary structural abnormalities. Clinical suspicion is the key to diagnosis, which is supported by establishing an environmental or occupational exposure to a causative inhaled antigen; by excluding infectious and other causes of recurrent, migratory, or progressive radiologic lung abnormalities; and by demonstrating antigen-specific antibodies in the serum. Two cases of hypersensitivity pneumonitis are described that illustrate the widely variable clinical, laboratory, and radiologic presentations and emphasize the importance of diagnosing this treatable condition. PMID- 8418321 TI - Intracardiac hydatid cyst: concise communication. AB - Hydatid cysts of the heart are rare and account for 0.5% to 2.0% of all hydatid infestations. The article describes a well-documented intracardiac hydatid cyst in a child. Because these cysts enlarge slowly, they rarely present in early childhood. PMID- 8418322 TI - Identification of internal mammary lymph nodes: value of the frontal chest radiograph. AB - The article describes eight patients with enlarged internal mammary lymph nodes visualized on the frontal plain chest radiograph. Enlarged internal mammary lymph nodes cast shadows that initially may be mistaken for a mediastinal or pleural abnormality. Although the lateral film alone may suggest these nodes, the findings on the frontal film help lateralize the abnormality. PMID- 8418324 TI - Case report: postoperative false aneurysm of the thoracic aorta demonstrated by helical scanning. AB - The article describes a case of postoperative false aneurysm of the thoracic aorta that was superbly demonstrated by helical computed tomography. This technique provides a simple method for obtaining rapid diagnostic studies in patients with this and similar conditions. PMID- 8418323 TI - Enlarging pleural effusion after liver transplantation. AB - Ten of 42 patients who underwent liver transplantation were retrospectively found to have enlarging pleural effusions later than 3 days after transplantation. Seven of the 10 patients had subdiaphragmatic pathology, including 4 with hematomas, 1 with a biloma, and 2 with abscesses. One patient with a subphrenic abscess also had an empyema. Patients with a enlarging pleural effusion later than 3 days after transplantation should be evaluated for subdiaphragmatic pathology. PMID- 8418325 TI - Talk to people about dying--they can handle it, say geriatricians and patients. PMID- 8418326 TI - National registry seeks scarce data on pregnancy outcomes during chemotherapy. PMID- 8418327 TI - Treatment of mild hypertension study shows results better when drugs abet life style changes. PMID- 8418328 TI - From the Food and Drug Administration. PMID- 8418329 TI - From the Centers for Disease Control and Prevention. Selected behaviors that increase risk for HIV infection, other sexually transmitted diseases, and unintended pregnancy among high school students--United States, 1991. PMID- 8418330 TI - From the Centers for Disease Control and Prevention. Update: influenza activity- United States and worldwide, 1992-1993 season. PMID- 8418331 TI - From the Centers for Disease Control and Prevention. Abortion surveillance: preliminary data--United States, 1990. PMID- 8418332 TI - From the Centers for Disease Control and Prevention. Unintended childbearing: pregnancy risk assessment monitoring system--Oklahoma. PMID- 8418333 TI - Back pain: the history and physical examination. PMID- 8418334 TI - Back pain: the history and physical examination. PMID- 8418335 TI - Back pain: the history and physical examination. PMID- 8418336 TI - Back pain: the history and physical examination. PMID- 8418337 TI - Back pain: the history and physical examination. PMID- 8418338 TI - Battered women: not-so-hot lines. PMID- 8418339 TI - Gender verification and the next Olympic games. PMID- 8418340 TI - California authorization of funds for the antismoking campaign. PMID- 8418341 TI - California authorization of funds for the antismoking campaign. PMID- 8418342 TI - Risk factors for hepatitis C virus seropositivity in heterosexual couples. AB - OBJECTIVES: To determine the risk of heterosexual transmission of hepatitis C virus (HCV) and to identify other risk factors for HCV seropositivity in heterosexual couples. DESIGN: Retrospective cross-sectional study comparing HCV seropositive and HCV-seronegative heterosexual men and women. SETTING: Couples recruited from the community and screened for participation in a study of the heterosexual transmission of human immunodeficiency virus. PARTICIPANTS: A total of 340 subjects, 170 men and 170 women in sexual partnerships, aged 18 through 61 years. MAIN OUTCOME MEASURE: Seropositivity for HCV antibodies. RESULTS: Overall, 31 (18%) of the 170 women and 56 (33%) of the 170 men were positive by a four antigen HCV immunoblot. Injection drug use and hemophilia were strongly associated with HCV seropositivity. Sixty-four percent of injection drug users were positive (odds ratio [OR], 27.0; 95% confidence interval [CI], 13.4 to 56.1; P < .0001), as were all four hemophiliacs in the study. History of blood transfusion was significantly associated with HCV seropositivity (OR, 2.7; 95% CI, 1.1 to 7.0; P = .02). Positivity for HCV was not associated with measures of sexual behavior within couples or with numbers of other sexual partners, history of sexually transmitted diseases, or human immunodeficiency virus seropositivity. However, two of the 31 women without parenteral risk but with a long-term HCV positive male partner were HCV seropositive compared with none of 81 women with an HCV-negative male partner (P = .07). CONCLUSIONS: These results provide little evidence of HCV sexual transmission but are consistent with infrequent sexual transmission. They corroborate the importance of injection drug use and transfusion of blood or blood products in transmitting HCV and underscore the importance of ascertaining parenteral exposures when examining sexual transmission of HCV. PMID- 8418343 TI - Relationship between malpractice claims and cesarean delivery. AB - OBJECTIVE: To investigate whether an association exists between the probability of a cesarean delivery and the level of malpractice claims risk faced by hospitals and physicians. DESIGN: Survey of computerized discharge data linked with physician and hospital malpractice claims records based on stratified, random sample of hospitals. SETTING: Acute care hospitals in New York State in 1984. POPULATION: All deliveries (60 490) at 31 hospitals. RESULTS: After controlling for the clinical risk of a cesarean delivery, patient socioeconomic status, and physician and hospital characteristics, cesarean delivery was positively associated with physician malpractice premiums (odds ratio [OR], 3.00; 95% confidence interval [CI], 2.13 to 4.24 for the difference between upstate and New York City levels), with the number of physician claims opened per 100 physicians at the hospital level (OR, 1.15; 95% CI, 1.02 to 1.30 for a 1-SD change), and with the number of hospital claims opened per 1000 discharges (OR, 1.26; 95% CI, 1.10 to 1.43 for a 1-SD change). Measures of physician-perceived risk of suit also showed a significant association with cesarean delivery (OR, 1.96; 95% CI, 1.53 to 2.52, upstate vs New York City). Within hospitals, there was no significant association (OR, 1.15; P = .126) between the odds of cesarean delivery and the claims history (none vs one or more) of an individual physician. CONCLUSION: Results support previous speculations of a positive association between malpractice claims risk and the rate of cesarean delivery. PMID- 8418344 TI - The impact of a regulation restricting medical house staff working hours on the quality of patient care. AB - OBJECTIVE: To examine the impact on patient care of a New York State regulation that restricted house staff working hours. DESIGN: Retrospective cohort study. SETTING: General medical service of an urban teaching hospital. PATIENTS: A total of 263 (94%) of 281 patients discharged from the study service during October 1988 and 263 (93%) of 283 patients discharged from the same service during October 1989. INTERVENTIONS: On July 1, 1989, New York State enacted a new regulation, Code 405, which limited residents' working hours and specified levels of supervision and ancillary support. MAIN OUTCOME MEASURES: In-hospital mortality, transfers to intensive care units, cardiopulmonary resuscitation attempts, discharge disposition, length of stay, medical complications, and house staff delays in ordering tests and procedures. RESULTS: Although the cohorts were comparable in severity of illness measures, more patients in 1989 suffered at least one medical complication (91 [35%] vs 59 [22%]; P = .002) and experienced at least one diagnostic test delay because of house staff (44 [17%] vs 4 [2%]; P < .001). These significant differences persisted after controlling for potential confounders in multivariate analyses. However, we found no significant differences in more serious outcomes: in-hospital mortality, transfer to intensive care unit, discharge disposition, or length of stay. CONCLUSION: These results suggest that restricted house staff working hours were associated with delayed test ordering by house staff and increased in-hospital complications. While these potentially deleterious effects on the quality of care did not result in statistically significant differences in more serious outcomes, further study at other hospitals is warranted to determine staffing strategies that optimize quality of care for patients, as well as medical education and quality of life for house officers. PMID- 8418345 TI - Physician inpatient order writing on microcomputer workstations. Effects on resource utilization. AB - OBJECTIVE: To assess the effects on health care resource utilization of a network of microcomputer workstations for writing all inpatient orders. DESIGN: Randomized controlled clinical trial. SETTING: Inpatient internal medicine service of an urban public hospital. SUBJECTS: A total of 5219 internal medicine patients and the 68 teams of house officers, medical students, and faculty internists who cared for them. INTERVENTION: Microcomputer workstations, linked to a comprehensive electronic medical record system, for writing all inpatient orders. MAIN OUTCOME MEASURES: Total inpatient charges for each admission and charges for specific categories of orders. A time-motion study of selected interns assessed the ordering system's time consumption. RESULTS: Intervention teams generated charges that were $887 (12.7%) lower per admission than did control teams (P = .02). Significant reductions (P < .05) were demonstrated separately for bed charges, diagnostic test charges, and drug charges. Reductions of similar proportion and statistical significance were found for hospital costs. The mean length of stay was 0.89 day shorter for intervention resident teams (P = .11). Interns in the intervention group spent an average of 33 minutes longer (5.5 minutes per patient) during a 10-hour observation period writing orders than did interns in the control group (P < .0001). CONCLUSIONS: A network of microcomputer workstations for writing all inpatient orders significantly lowered patient charges and hospital costs. This would amount to savings of more than $3 million in charges annually for this hospital's medicine service and potentially tens of billions of dollars nationwide. However, the system required more physician time than did the paper charts. Research at other sites and system advances to reduce time requirements are warranted. PMID- 8418346 TI - The changing epidemiology of invasive group A streptococcal infections and the emergence of streptococcal toxic shock-like syndrome. A retrospective population based study. AB - OBJECTIVE: To determine disease incidence and changes in the epidemiology of invasive group A streptococcal infections in a community in Arizona. DESIGN AND SETTING: We retrospectively surveyed microbiology records from all 10 hospitals in Pima County, Arizona, to identify patients who had Streptococcus pyogenes isolated from blood, sterile body fluid, or tissue biopsy specimens between April 1985 and March 1990. Demographic and clinical information was abstracted from the medical records of these patients. PATIENTS: A total of 128 patients with a median age of 53.5 years (range, 6 months to 96 years). OUTCOME MEASURES: Racial/ethnic differences in disease incidence; mortality and changes in the clinical spectrum of disease over the study period. RESULTS: The annual age adjusted incidence was 4.3 per 100,000 but was 46.0 per 100,000 among Native Americans. Advanced age, age less than 5 years, hypotension, and multi-organ system involvement were significantly associated with increased mortality. From 1985 to 1990, the proportion of infections with hypotension, rash, desquamation, renal impairment, and gastrointestinal involvement increased significantly (chi 2 for trend P < or = .02 for each feature). A toxic shock-like syndrome occurred in 8% of infections since 1988, compared with none of the infections between 1985 and 1987 (P = .04). Patients with the syndrome were younger than patients with other invasive infections (median age 15 vs 54 years, P = .02), and were less likely to have underlying medical conditions (P = .008). CONCLUSIONS: Significant changes occurred in the spectrum of invasive group A streptococcal infections in Pima County, Arizona, between 1985 and 1990. Native Americans were at increased risk of acquiring these infections. Patients with the streptococcal toxic shock like syndrome had epidemiologic features that distinguished them from patients with other invasive infections, including younger age and less underlying illness. PMID- 8418347 TI - Defining the group A streptococcal toxic shock syndrome. Rationale and consensus definition. The Working Group on Severe Streptococcal Infections. PMID- 8418348 TI - Hepatitis C virus infection among patients attending a clinic for sexually transmitted diseases. AB - OBJECTIVE: To evaluate the association between hepatitis C virus (HCV) infection and sexual behavior in a sexually active population. DESIGN: Cross-sectional study. SETTING: Inner-city clinic for sexually transmitted diseases. SUBJECTS: The study included 1292 patients attending the clinic for care during a 1-month period and having syphilis serologic tests performed. OUTCOME MEASURES: Antibody to HCV (anti-HCV) positivity as defined by a repeatedly-reactive enzyme immunoassay and a positive neutralization enzyme immunoassay (Abbott Laboratories, Chicago, Ill). RESULTS: Of 1292 patients screened for anti-HCV, 99 (7.7%) were positive. Logistic regression analysis found that patients who reported intravenous drug use, were positive for antibody to hepatitis B core antigen, reported a history of a blood transfusion, were black, or reported crack cocaine use were more likely to be anti-HCV-positive. Forty-five percent of patients who were anti-HCV-positive reported intravenous drug use. Sex with an intravenous drug user and a history of gonorrhea and syphilis were associated with anti-HCV positivity in a univariate analysis, but after controlling for confounding variables, no such associations remained. While having multiple sexual partners in the previous 3 months, being homosexual or bisexual, and engaging in receptive anal intercourse were associated with being positive for antibody to hepatitis B core antigen, those behaviors were not associated with anti-HCV positivity. CONCLUSIONS: While these results cannot exclude a role for the sexual transmission of HCV, they do suggest that, in this sexually active population, the sexual transmission of HCV occurs infrequently and that HCV is largely associated with intravenous drug use. PMID- 8418349 TI - Elective cyclosporine withdrawal after renal transplantation. A meta-analysis. AB - OBJECTIVE: To determine whether it is safe to electively discontinue cyclosporine therapy after renal transplantation. DATA SOURCES: MEDLINE and bibliographies from recent publications. STUDY SELECTION: Controlled trials assessing the rate of acute rejection, graft loss, and mortality after elective cyclosporine withdrawal. DATA EXTRACTION: We compared outcomes in patients who underwent withdrawal from cyclosporine treatment with patients who were not withdrawn (part 1), and in a separate analysis (part 2), with patients who never received cyclosporine. DATA SYNTHESIS: In part 1 of the meta-analysis, consisting of 10 randomized and seven nonrandomized trials, there was a greater combined rate of acute rejection among patients in whom cyclosporine was withdrawn compared with control patients who continued to receive cyclosporine (weighted difference in episodes per patient, 126; 95% confidence interval [CI], 0.085 to 0.167; P < .001). However, there were no differences in graft loss (weighted difference in grafts lost per patient per year, -0.009; 95% CI, -0.022 to 0.004; P = 0.19) or mortality (weighted difference in deaths per patient per year, -0.005; 95% CI, 0.016 to 0.006; P = .40) attributable to cyclosporine withdrawal. In part 2 of the meta-analysis, consisting of three randomized and three nonrandomized trials, the combined rate of graft loss for patients who were withdrawn from cyclosporine was not significantly different vs control patients who never received cyclosporine (weighted difference in grafts lost per patient per year, -0.020; 95% CI, -0.043 to 0.003; P = .08). However, when the three randomized trials were analyzed separately, graft survival was better in patients who were withdrawn from cyclosporine (weighted difference in grafts lost per patient per year, 0.0382; 95% CI, 0.0002 to 0.0762; P = .049). None of the outcomes was affected by the timing or manner of the cyclosporine withdrawal. CONCLUSIONS: The increased incidence of acute rejection following elective cyclosporine withdrawal does not affect short-term graft or patient survival after renal transplantation. Whether long-term consequences will outweigh the benefits of elective withdrawal remains to be determined. PMID- 8418351 TI - Supervision, not regulation of hours, is the key to improving the quality of patient care. PMID- 8418350 TI - Ethical considerations in listing fetuses as candidates for neonatal heart transplantation. PMID- 8418352 TI - A piece of my mind. Social ... Security. PMID- 8418353 TI - [A clinicopathologic study of thymomas: a review of 52 cases, with particular reference to results of treatment]. AB - Clinicopathologic aspects of 52 thymomas were reviewed. Masaoka staging revealed stage I in 32, stage II in 2, stage III in 12, stage IV a in 3, and stage IV b in 3. Eleven patients (21%) had associated myasthenia gravis (MG), and three (6%) had pure red cell aplasia (PRCA). Operation was complete resection in 39 patients (stage I 32, stage II 2, stage III 2, and stage IV b 3), subtotal resection in 4 patients (stage III 2, stage IV a 2), partial resection or simple biopsy in 7 patients (stage III 6, stage IV a 1). Survival rates at 5 years and 10 years were 88%, 88% in noninvasive thymomas, and 65%, 45% in invasive thymomas, respectively. Statistical analysis indicated that patients with noninvasive thymomas had significantly longer survival than those with invasive thymomas. Significant difference was not found in survival on the basis of tumor cell type according to lymphocyte/epithelial cell ratio. Associated MG had no influence on the survival of thymoma patients, however, the presence of PRCA was an adverse factor in survival. In cases of invasive thymomas, there was statistically significant difference in the survival rates between the group of patients undergoing complete or subtotal resection and the group undergoing partial resection or only biopsy. Irradiation was of value in local control of thymoma, but lymphogeneous or hematogeneous recurrence occurred in 21% of patients with invasive thymomas from 3 months to 12 years postoperatively. Lifelong follow-up should be necessary in thymoma patients since late recurrence is not so rare. PMID- 8418354 TI - [Surgical therapy with adjuvant radiotherapy of thymoma: results of 104 cases]. AB - One hundred and four cases of thymomas were treated between 1967 and 1991 at the Chest Disease Institute, Kyoto University. Forty-two cases were clinical stage I, 12 cases were stage II, 42 cases were stage III and 8 cases were stage IV. Nineteen patients had MG, 17 patients had SVC syndrome. Ninety-six cases were surgically resected combined with radiation therapy. Overall survival was 77.9% at 5 years and 62.5% at 10 years. The survival of 62 cases of invasive thymoma was 77.5% at 5 years and 54.8% at 10 years. We've got better prognosis when total or subtotal resection of invasive thymoma could be accomplished, i.e. 88.8% at 5 years and 72.6% at 10 years. PMID- 8418355 TI - [Multidisciplinary treatment of advanced invasive thymoma]. AB - From February 1988 to September 1992, six patients with Stage III or IV invasive thymoma were enrolled onto our study of multidisciplinary treatment, which included chemotherapy, surgery, and radiotherapy. The chemotherapy consisted of cisplatin (20 mg/m2 continuous 24-hour infusion on days 1 through 4), doxorubicin (40 mg/m2 on day 1), and methylprednisolone (1,000 mg on days 1 through 4 and 500 mg days 5 and 6). Two patients received surgery (incomplete resection) and postoperative chemo/radiotherapy, and four patients received induction chemotherapy followed by surgery and chemo/radiotherapy. All patients with induction chemotherapy showed a partial response and underwent complete (n = 2) or partial (n = 2) resection. Four patients are alive with no evidence of disease for 31 to 55 months after the diagnoses. But two with partial resection after chemotherapy died of recurrent disease. Multidisciplinary treatment including chemotherapy may be a new therapeutic choice in management of advanced invasive thymoma. PMID- 8418357 TI - [Surgical treatment of stage IVa thymoma]. AB - The relationship between prognosis of the disease and the type of surgery were reviewed in 110 cases of thymoma. Surgical procedures were classified into four types: total resection (TR), subtotal resection (SR), partial resection (PR) and exploratory thoracotomy (ET), and the extent of the disease was expressed as stage I through IV based on Masaoka's classification. Ten-year survival rates for stage III and IVa diseases were 5 6.2% in TR, 37.9% in SR and 30.3% in PR. None of the patients in ET survived more than 28 months. Tumor death after surgery was not observed in TR. Two of three patients with stage IVa disease who underwent TR are alive without recurrence of the tumor at 37 months and 6 months after surgery. The pleuropneumonectomy was performed in the former case. One of four patients who underwent SR and one of three patients who underwent PR both with stage IVa disease are alive at 71 and 150 months after surgical intervention, respectively, in spite of the presence of the tumor relapse. In these patients, repeated chemotherapy and radiotherapy were administered following the resection of the primary lesion. In conclusion, it was indicated that the improvement of the survival rate could be obtained by resection the tumor as completely as possible in stage III and IVa thymoma, as long-term survival has been proved to be possible by macroscopically complete removal of the primary site and pleural disseminations followed by combined adjuvant therapy in patient with stage IVa disease. PMID- 8418356 TI - [Treatment of invasive thymoma using low dose and extended-field irradiation including hemi-thorax or whole-thorax]. AB - The records of 10 patients treated for invasive thymoma with low-dose and extended-field irradiation including hemi-thorax or whole-thorax are reviewed. Six patients were in stage III. They were treated with up to 15 Gy hemi-thorax irradiation which was followed by shrinkage of the radiation field after removal of tumor. Five of them have lived 4 to 10 years, while one remaining patient died of uncertain cause. Four patients had stage IVa thymoma. Two patients in an earlier stage were treated with irradiation alone; whole-thorax irradiation of up to 20 Gy followed by local irradiation. Those two patients, however, developed severe pulmonary infection secondary to irradiation pneumonitis. Furthermore, local relapse was observed in one patient who died of respiratory insufficiency after 6 years. In the remaining 2 patients in stage IVa, intensive chemotherapy was administered before whole-thorax irradiation of up to 15 Gy. One of these patients is alive with no evidence of recurrence for 5 years, while the other died of a disease not related to thymoma. These facts indicate that whole-thorax irradiation combined with chemotherapy may be of value in preventing local relapse with stage IVa thymoma. PMID- 8418358 TI - [Surgical treatment of thymoma--clinical evaluation]. AB - Sixty-seven patients with thymoma were surgically treated during the past 19 years in our department. The 5-year, 10-year and 15-year survival rates of total cases with thymomas were 69.2%, 59.6% and 59.6%, respectively. Survival rates of thymoma with MG and without MG were not significantly different. According to clinical stages in Masaoka's classification, there were significant difference between Stage I and Stag III (p < 0.05), Stage I and Stage IV a (p < 0.01), and Stage I and Stage IV b (p < 0.01). We can conclude that complete resection of thymomas lead to better prognosis, and immunochemotherapy using cyclophosphamide, vincristine and OK-432 are effective. PMID- 8418359 TI - [Therapy of stage IV thymoma]. AB - The clinical and pathological features of three patients with stage IV thymoma were presented. These patients were classified into stage IV at the first admission according to pericardial dissemination, mediastinal lymph node metastasis, and lung metastasis, respectively. One patient was accompanied with myasthenia gravis. All three patients underwent operations for primary thymoma. The metastatic lesions were also resected as completely as possible. Although postoperative radiotherapy was employed with all patients at the first therapy, chemotherapy was administered to 2 cases. After the first therapy, two patients had pleural dissemination and bilateral multiple lung metastases. They underwent second operations for pleural dissemination and metastatic lung tumors, and postoperative chemotherapy was performed. All three patients are alive (ranged, 2 years 6 months to 4 years 4 months postoperatively). Although follow-up of these three patients is short-term, it seems likely that radiotherapy and chemotherapy following surgery are effective treatments for patients with stage IV thymoma. PMID- 8418361 TI - [Surgical treatment of myasthenia gravis in patients with thymoma]. AB - During the 18 years period from 1972 to 1990 fifty-two patients with thymoma, including 13 with thymoma and myasthenia gravis (MG), underwent surgical treatment in our hospital. During the same period 27 patients without thymoma underwent surgical intervention for MG. These patients were assessed from the clinical manifestations, clinical effects for MG, and prognosis. The improved effects of surgery for MG with or without thymoma could be yearly obtained after operation. Palliation and remission rates of the 10 patients of MG with thymoma, excluded postthymectomy MG, were very poor in comparison with that of the 27 patients of MG without thymoma. Further more, the prognosis of the 13 patients of thymoma with MG was worse than that of 39 patients of thymoma without MG. The association of thymoma and progressive severity of disease related closely the influence of delayed response to surgical intervention and prognosis. In conclusion, aggressive surgical resection followed by chemo-radiotherapy was the best treatment for the invasive thymoma. Postoperative intensive care for MG crisis and recurrence of thymoma improved the effect to surgical intervention and revealed the good prognosis. PMID- 8418360 TI - [Management of myasthenia gravis (MG) associated with thymoma]. AB - From June 1975 to June 1992, we experienced 203 patients with MG. Sixty patients had associated thymoma, registering stages I (n = 31), II (n = 18), III (n = 9), and IV a (n = 2), according to the classification of Masaoka and colleagues. Fifty patients had generalized MG and 10 had ocular MG. Histopathological findings indicated round-oval, polygonal cell type in 46 patients and mixture of round-oval and spindle cell type in 11 patients and invasive thymomas had a tendency to have a predominantly epithelial type with and increased epithelial element as compared with non-invasive thymomas. An extended thymectomy including thymoma was performed in all patients. Three patients in stage II, 7 patients in stage III, and 2 patients in IV a received postoperative radiation therapy. Twenty-one patients needed prolonged respiratory care for respiratory crisis. Fifty-three patients have been doing well with the alternate-day corticosteroid therapy, however, 4 patients had an excavation of myasthenic symptoms associated with recurrence of thymoma on the pleura. In conclusion, early extended thymectomy including thymoma is markedly effective therapy for MG with thymoma and a careful attention should be paid for recurrence in the patients with invasive thymoma. PMID- 8418362 TI - [The treatment of thymoma with myasthenia gravis: report of 20 cases]. AB - Sixty-eight patients with myasthenia gravis underwent thymic surgery in our department. Among of 68 cases, twenty patients with thymoma were followed and their prognosis were investigated. Sixteen cases received a extended thymectomy with thymomectomy, six cases received local irradiation, and 16 cases received a chemotherapy. Among of 16 cases, 13 cases received corticosteroid only, and 3 cases received CHOP (CPA+ADM+VCR+PSL). Survival rates for thymoma of 20 cases were 94.7%, 88.9%, 78.6%, and 54.5% at 1, 3, 5 and 7 years, respectively. Survival rates for thymoma at 5 years were 83.3%, 50.0%, 100%, and 100% in stage I (complete encapsulation of 8), II (invasion into pericapsular fatty tissue of 7), III (invasion into surrounding organ of 3), and IV a (pleural dissemination of 2), respectively. Survival rate for extended thymectomy with thymomectomy of 16 cases was 81.8% at 5 years. Among of 18 total resection cases of thymoma, a recurrence occurred (5.6%). Causes of 6 death after surgery were crises of myasthenia gravis (3), other diseases (2), and recurrence of thymoma (1). PMID- 8418363 TI - [A case of malignant thymoma associated with myasthenia gravis, pure red cell aplasia and high serum level of SCC antigen]. AB - We present a case of 43-year-old man with pure red cell aplasia appearing 8 years after thymothymectomy. He underwent an operation and postoperative radiotherapy for malignant thymoma associated with myasthenia gravis in 1983. In 1991 high serum level of SCC antigen was noted and a metastatic tumor was found in the retroperitoneal region. After removal of the tumor pure red cell aplasia developed, which responded to steroid therapy. The serum level of SCC antigen is still abnormally high. PMID- 8418364 TI - [Reoperation in patients with thymoma]. AB - For past seven years, reoperations were carried out for six patients with thymoma and myasthenia gravis (MG). Of six patients, five patients were suspected recurrent thymoma and remaining one patient was diagnosed as the residual thymus after thymothymectomy or thymomectomy. Myasthenia gravis has been progressing in five patients after initial operation except for one patient. Two cases of pleural dissemination tumors, one case of local recurrent thymoma and one case of the residual thymus were confirmed by surgery, remaining two patients had no recurrent tumor. As surgical procedure, median sternotomy using for reoperation has a great surgical risk, because left brachiocephalic vein closely adhered to the sternum is apt to be injured. Clinical symptoms of MG were improved in all the patients after reoperation regardless of recurrent tumor. PMID- 8418365 TI - [Flow cytometric analysis of the DNA content in resected thymomas]. AB - Nuclear DNA contents were determined by flow cytometry in 31 resected thymomas. Ten (32%) of the 31 thymomas showed DNA aneuploid, and the presence of aneuploidy was well correlated with advanced clinical stage. Patients with an aneuploid tumor had a poor prognosis than those with a diploid tumor, demonstrating a survival rate of 50% at 7 years in aneuploidy compared to 100% at the same period in diploid patients (p < 0.05). In addition, patients with a high DNA index (DI > or = 1.5) tended to have a poor prognosis than those with a low DI. The analysis of DNA content can be an important prognostic index in patients with a thymoma. PMID- 8418366 TI - [Tumor-node-metastasis classification for thymic epitherial tumors]. AB - We applied the tentative TNM classification of thymoma to 24 cases of IV b thymoma, 59 cases of thymic carcinoma, and 26 cases of thymic carcinoma (109 cases in total). This classification was useful in these cases because cases could be divided into different grades (e.g. N 0, N 1, N 2, N 3) of the factor N (lymph node metastasis) without any shift to particular grades. Based on this finding, we hereby propose a TNM classification of thymic epitherial tumors which was prepared by slightly modifying the tentative TNM classification of thymoma. PMID- 8418367 TI - [Aortic dissection]. PMID- 8418368 TI - [Surgical treatment of thymomas]. AB - Forty-eight patients were evaluated. These cases were classified to stage I in 17, stage II in 11, stage III in 13, and stage IV in 7 by Masaoka staging. In 43, a complete resection was performed and in 5, an incomplete resection was carried out. Twelve patients between stage III and IV received postoperative radiotherapy because of incomplete resection and advanced invasive case (defined as high-risk group). Survival at 10 years was 55% for patients who were in high-risk group. This result was disappointed because survival was low in spite of surgical resection combined with postoperative radiation. From that reason, more concentrated therapy might be necessary with not only enlargement of radiation field but also chemotherapy for vascular invasive thymoma in addition to resection. PMID- 8418369 TI - Collaborative study of karyotypes in childhood acute lymphoblastic leukemias. Groupe Francais de Cytogenetique Hematologique. AB - A collaborative study carried out by the Groupe Francais de Cytogenetique Hematologique collected 411 successful karyotypes of childhood acute lymphoblastic leukemias. Karyotypes showed a clonal abnormality in 292 patients (71%). The distribution of ploidy groups was: pseudodiploidy in 116 karyotypes (28.2%), hyperdiploidy > 50 chromosomes in 110 karyotypes (26.8%), hyperdiploidy 47-50 chromosomes in 46 karyotypes (11.2%) and hypodiploidy in 20 karyotypes (4.9%). One-half of the patients with hyperdiploidy > 50 chromosomes also had a structural abnormality, with a partial trisomy 1q in one fourth of them. Similar translocations, candidate for new recurrent changes were identified: t(9;9)(p13;q13),t(7;9)(q11;p11),t(7;12)(q11;p12-13), t(4;12)(q13;p12), and t(1;17)(q12-21;p13). Within recurrent translocations, the three t(10;11)(p13 14;q14-21) displayed a T-cell phenotype. In T-cell leukemias, a new area of recurrent breakpoints (5q31-35) was observed and deletions 6q were more frequent in this lineage. Correlations of cytogenetic results with clinical and hematological data revealed that, within hyperdiploidy > 50 chromosomes, patients with structural changes were older than patients without. Patients with 9p changes showed some of the features usually observed in lymphomatous leukemias. Even with a short follow-up, differences in outcomes were observed. Patients with hyperdiploidy > 50 chromosomes fared the best and those with pseudodiploid karyotypes did worse than patients with other karyotypes. Patients with random translocations did not share the poor outcome of patients with recurrent translocations. PMID- 8418370 TI - Sensitive and reproducible detection of occult disease in patients with follicular lymphoma by PCR amplification of t(14;18) both pre- and post treatment. AB - The t(14;18) chromosomal translocation occurs in most follicular non-Hodgkin's lymphomas and places the Bcl-2 gene on chromosome 18q21 into the immunoglobulin JH region on chromosome 14q32. This translocation can be exploited to detect clonal malignant cells bearing this genetic alteration. A polymerase chain reaction (PCR) assay amplifying over the major breakpoint region (mbr) and minor cluster region (mcr) was developed and optimized. In this report, the sensitivity and reproducibility of this semiquantitative assay, performed on a relatively large number of clinical samples is shown. A titration curve of DNA made from a t(14;18)- cell line admixed with increasing ratios of a t(14;18)+ cell line was used to demonstrate that one t(14;18)+ cell in 100,000 t(14;18)- cells could reproducibly be detected. Occult lymphoma cells, not detected by standard morphologic analysis, were demonstrated in almost two-thirds of the bone marrow and peripheral blood specimens obtained from untreated patients with follicular lymphoma. Of 11 bone marrow samples assessed, seven were positive for occult disease by PCR amplification over the mbr and one was positive over the mcr. Of these six positive marrow samples, only three had been reported positive by standard morphologic criteria. In addition, seven of nine peripheral blood samples assessed were positive over the mbr and one additional sample was positive over the mcr. None of these were morphologically positive. Seven of the above patients would have been upstaged if these results were utilized for staging, including two of three patients with stage I or stage II disease. PCR detectable occult disease persisted in four of four patients assessed both pre- and post-treatment, even after aggressive multi-drug combination chemotherapy in two of these patients. The clinical significance of detecting this occult disease must await the study of larger numbers of patients and the clinical outcomes of patients with occult disease and patients without occult disease. PMID- 8418371 TI - Wheat germ agglutinin affinity of murine hemopoietic stem cell subpopulations is an inverse function of their long-term repopulating ability in vitro and in vivo. AB - Hemopoietic stem cells show extensive heterogeneity with respect to their proliferative potential and activity. We have recently reported that the accepted technique for sorting stem cells on the basis of high affinity for the lectin wheat germ agglutinin (WGA) did not select for cells initiating long-term production of new stem cells on a stromal layer in vitro. We have therefore reinvestigated the expression of cell surface sialic acid residues in the hemopoietic stem cell compartment by sorting murine bone marrow cells on the basis of affinity for WGA. Frequency analysis of long-term bone marrow culture initiating stem cells was done using the cobblestone-area-forming cell (CAFC) assay with limiting dilution set-up. In vivo stem cell quality was determined by spleen colony formation, marrow-repopulating ability (MRA) and long-term repopulating ability (LTRA) using sex-mismatched hemopoietic chimerism. The data indicate that MRA and LTRA in vivo and in vitro are among the most WGA-dim cells. In contrast, the enrichment factors for splenic colony-forming units (CFU-S) at day 12 and transient CAFC increase with increasing WGA affinity. These characteristics allowed us to concentrate LTRA cells 590- to 850-fold over their activity in normal bone marrow without significant enrichment of day-12 CFU-S. The data reveal that WGA affinity is an inverse function of the primitiveness of murine hemopoietic stem cells and that long-term production of blood cells in vivo and in vitro is provided for by primitive cells that are physically separable from the vast majority of day-12 CFU-S. In addition the data reveal, that the CAFC frequency at day 28-35 of a graft strongly correlates with the number of cells required to induce 40% donor-type chimerism at 15 months post transplantation and thus predicts the in vivo LTRA of a graft. PMID- 8418372 TI - Retrovirus-mediated transfer and expression of marker genes in the BN rat acute myelocytic leukemia model for the study of minimal residual disease (MRD). AB - To study minimal residual disease (MRD) in leukemia, we transferred the Escherichia coli genes encoding beta-galactosidase (lacZ) and neomycin resistance (neo(r)) into the subline LT12 of the Brown Norway rat acute myelocytic leukemia (BNML), employing the retroviral BAG vector. In this way leukemic cells were genetically marked. Ten independent cell lines were characterized during in vitro growth as well as during two subsequent in vivo passages for expression of neo(r) for which the neomycin analogue G418 was used, and for lacZ expression for which the substrate 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-gal) was used. Out of 10 lines, four revealed permanent high expression of lacZ in all cells. In four other lines greatly varying lacZ expression between the individual cells from these lines was observed. In the remaining two lines lacZ expression was gradually lost. In contrast, neo(r) expression was gradually lost in eight out of the 10 lines, particularly rapidly during in vivo passaging. In the remaining two lines neo(r) expression was retained. The genetic modification did not alter the in vitro leukemogenicity of the cells. Long term in vivo expression of neo(r) and lacZ was followed in two selected lines up to 12 subsequent passages, i.e. one from the group of homogeneous high lacZ expression and one from the group of heterogeneous lacZ expression. In both lines lacZ expression was retained whereas neo(r) expression was rapidly lost after the third passage. The feasibility of using genetically marked leukemic cells for studies of minimal residual disease (MRD) was explored by injecting rats with leukemic cells, treating them with chemotherapy at full blown leukemia development to reduce the tumor load, mimicking the induction of a state of MRD and studying lacZ expression at relapse. LacZ expression was evident in 100% of the cells whereas neo(r) expression was lost in a considerable fraction. These results indicate that the viral vector BAG can be used to mark leukemia cells genetically although a selection of clones with the desired stability of long-term expression is required. PMID- 8418373 TI - Demonstration of chimerism after allogeneic bone marrow transplantation by polymerase chain reaction of Y-chromosome-specific nucleotide sequences- characterization of a new technical approach. AB - Various techniques are applied to assess chimerism after allogeneic bone marrow transplantation. The polymerase chain reaction (PCR) of donor- and/or host specific gene sequences provides a rapid and highly sensitive technique. We describe the characterization and application of PCR for the amplification of Y chromosome-specific DNA in blood cells recovered from stored slides. Four different primer pair combinations were used. PCR can be rapidly performed on stained or unstained slide material with varying sensitivity--depending on the primer combination. The lowest limit of detection is one male cell in 500-1000 female cells. The technique was applied to follow the early post-transplant course of 15 male patients who received grafts from female donors and found a high incidence of mixed chimerism during the first three months after BMT and a striking fluctuation between positive and negative results in the follow-up of individual patients. We conclude that PCR for the detection of male-specific DNA sequences can be successfully performed with high sensitivity on material recovered from stored blood slides. PMID- 8418374 TI - Hypothesis: apoptosis may be the mechanism responsible for the premature intramedullary cell death in the myelodysplastic syndrome. PMID- 8418375 TI - Coagulation disorders associated with acute promyelocytic leukemia: corrective effect of all-trans retinoic acid treatment. AB - The bleeding diathesis in patients with acute promyelocytic leukemia (APL) is generally attributed to disseminated intravascular coagulation (DIC), initiated by the release of procoagulant activity from leukemic cells. Primary fibrinogenolysis, mediated by the release of leukocyte proteases, may also contribute to this disorder. We analyzed coagulation parameters in 15 non-septic APL patients. Before treatment, there was evidence of thrombin activation with DIC: increased levels of circulating thrombin-antithrombin III complexes, prothrombin fragments 1 + 2 and D-Dimer complexes. This DIC syndrome was probably limited, since no prothrombin time decrease, no significant factor V consumption, and normal levels of coagulation inhibitors (antithrombin III and protein C) were observed in APL patients when compared to normal controls. In this context, marked hypofibrinogenemia suggested primary fibrinogenolysis as the predominant etiology. Despite normal or high tissue plasminogen activator (tPA) and plasminogen activator inhibitor (PAI-1) antigen levels, the plasma PAI-1 activity and the formation of tPA/PAI-1 complexes were lower in APL patients than in normal controls, suggesting a proteolytic degradation of PAI-1, not able to complex tPA. Two other fibrinolytic inhibitor molecules (alpha-2 plasmin inhibitor antigen and histidine-rich glycoprotein antigen) were also significantly reduced, as well as the two subunits of fibrin stability factor XIII, although only subunit A is known to be susceptible to thrombin action. Evidence of degraded forms of von Willebrand factor in the plasma suggested an extended proteolytic activity. Four patients treated with all-trans-retinoic acid (ATRA) as a single differentiating agent were studied serially. A dissociation between these two syndromes--DIC and fibrinogenolysis/proteolysis--was observed. The rapid correction of the lysis markers contrasted with a more prolonged persistence of the procoagulant activity. We observed persistently high elastase/alpha 1-proteinase inhibitor complex levels during ATRA therapy, despite progressive correction of all lysis markers. Thus, the release of elastase from promyelocytic leukemic cells is probably not the only determinant of the fibrinogenolytic/proteolytic syndrome. In summary, the present findings provide new arguments for the association of DIC and proteolysis syndromes in APL associated coagulation disorders. Further prospective studies are needed in order to confirm the persistence of thrombin activation in course of ATRA therapy. PMID- 8418378 TI - Expression of activation antigens CD38 and CD71 is not clinically important in childhood acute lymphoblastic leukemia. AB - Cell activation antigen expression, because it is related to cell proliferation, may offer useful information about tumor characteristics and treatment outcome. To assess the clinical utility of assays for the plasmocyte activation antigen CD38 (T10) and the thymocyte activation antigen CD71 (transferrin receptor; T9) in childhood acute lymphoblastic leukemia (ALL), we reviewed the presenting features and clinical outcomes of 325 ALL patients treated on a single protocol at St Jude Children's Research Hospital. T-cell ALL cases were more likely than B lineage ALL cases to express CD38 (100% versus 90.5%, p < 0.01) and CD71 (92% versus 32%, p < 0.01). However, expression of these antigens was not related to any clinical or biologic feature within the immunophenotypic subgroups. More importantly, event-free survival did not differ significantly between CD38+ and CD38- cases or between CD71+ and CD71- cases in the study group as a whole or in immunophenotypic subgroups. Studies of CD38 and CD71 expression in childhood ALL provide no clinically useful information beyond that obtained from standard immunophenotyping studies. PMID- 8418376 TI - Geographical distribution of acute lymphoblastic leukaemia subtypes: second report of the collaborative group study. AB - Childhood acute lymphoblastic leukemia (ALL) T and B precursor subtypes have been identified by standardised immunophenotyping in different geographic and ethnic settings. Comparison of the relative frequencies and estimated incidence rates of the major subtypes indicates very similar values, with the striking exception of black childhood populations in Africa in which there appears to be a significant and selective deficit in the incidence of the common (B-cell precursor) subset of ALL. There is suggestive evidence for a similar bias in ALL subtypes in South Africans of mixed ethnic origin and in Mapuche Indians from Chile. Several interpretations of these data are possible but the one favoured attributes these differences primarily to socio-economic factors and patterns of infection in infancy. PMID- 8418377 TI - Clinical significance of CD10 expression in childhood acute lymphoblastic leukemia. AB - The independent significance of CD10 expression in childhood acute lymphoblastic leukemia (ALL) is uncertain because most studies have not adjusted for other risk features, such as age and immunophenotype, or for treatment effects. We reassessed the clinical importance of CD10 expression in patients who received highly effective contemporary treatment. CD10 antigen was detected in blast cells from 384 of 408 patients (94%) with B-lineage ALL and 36 of 90 (40%) with T-cell ALL. In the B-lineage subgroup, CD10 expression was associated with favorable presenting features: age > or = 1 year, lower leukocyte count (< 50 x 10(9)/l), and leukemic cell DNA index > or = 1.16 or hyperdiploidy > 50 chromosomes. One half of the patients with CD10- B-lineage ALL had 11q23 chromosomal abnormalities. Separate analysis of the marker in T-cell ALL revealed no differences between CD10+ and CD10- cases in clinical features or karyotypic patterns, with the exception of a lower frequency of central nervous system leukemia and a higher frequency of 9p abnormalities in the former subgroup. CD10+ T-cell cases were also significantly more likely than CD10- cases to coexpress CD21, CD1, CD4, or CD8. Lack of CD10 expression was independently associated with an adverse prognosis in T-cell ALL (p = 0.02). However, for the larger subgroup of patients with B-lineage ALL, CD10 expression has no independent prognostic significance. PMID- 8418379 TI - Microtubule network and microtubule-associated proteins in leukemic T lymphocytes. AB - Cytoskeletal changes have been known to occur in cell transformation. Vinca alkaloids which bind to the tubulin dimer and inhibit microtubule (MT) assembly as well as disrupting the MT network and mitotic spindle, have been used as cancer chemotherapeutic agents. It has been proposed that apart from their anti mitotic activity, these drugs act on the peripheral MT of malignant cells to produce their cytolytic effects. In this paper we demonstrate the presence of an altered cytoplasmic MT network in MOLT-4 and HuT-78 leukemic cells (human T-cell leukemic lines) compared to normal human peripheral blood lymphocytes stimulated with mitogens. In addition, using a selective extraction protocol we have compared microtubule-associated proteins (MAPs) profiles of G1/S synchronized leukemic human T-cells and 20 h mitogen-stimulated human peripheral blood T cells. We observed a dramatic decrease in the expression of a MAP of apparent molecular weight 52 kDa and pI 5.2 in the leukemic cells synchronized at the G1/S border of the cell cycle. These results suggest that altered MT network morphology and MAP synthesis may be components of the malignant phenotype in the T-lymphocytic leukemias studied here. PMID- 8418380 TI - Expression of the monocyte-specific esterase gene in leukemia-lymphoma cell lines. AB - The expression of the monocyte esterase was examined in a panel of 77 continuous human leukemia-lymphoma cell lines representing all hematopoietic cell lineages and in 16 other cell lines. Accumulation of mRNA, determined by Northern blotting with the cDNA probe HMSE-1, and production of the protein, shown by isoelectric focusing on polyacrylamide gels, correlated with differentiation of the cells along the monocytic lineage. None of the lymphoid, erythroid, megakaryoblastic or Hodgkin's disease derived cell lines or the non-hematopoietic human tumor cell lines expressed the full-length mRNA of 2.0 kb. These results support the notion that this enzyme, a serine hydrolase with still unknown physiological functions, is specifically expressed in cells committed to the monocyte/macrophage cell lineage. PMID- 8418381 TI - Structure and expression of the GM-CSF receptor alpha and beta chain genes in human leukemia. AB - In this paper we report on the structure and expression of the genes encoding the alpha and beta chains of the granulocyte-macrophage colony-stimulating factor (GM CSF) receptor in human leukemia. The alpha chain gene is highly polymorphic in normal individuals and no evidence for rearrangement within this locus was found in 47 hemopoietic, nine non-hemopoietic malignancies and five human cell lines. Using the polymerase chain reaction the gene for the alpha chain was shown to be expressed in 18/18 primary myeloid as well as 8/8 primary lymphoid leukemias analysed. To investigate the integrity of the mRNA, polymerase chain reactions (PCR) using a combination of oligonucleotides spanning the entire coding region of the alpha chain were performed. Normal sized fragments were generated with all combinations of oligonucleotides from all but one leukemia. One chronic lymphoid leukemia displayed an apparent alteration at the 3' end of the 3' untranslated region of the alpha chain mRNA. No polymorphisms were detected in the beta chain gene which was also not rearranged in any of the samples analysed. The beta chain mRNA was expressed in 17/18 primary myeloid and 7/8 primary lymphoid leukemias and in those leukemias there was no evidence for any lesions in the mRNA, as judged by PCR fragment size. Thus gross structural lesions in the genes encoding the GM-CSF receptor alpha and beta chains appear to be infrequent in hemopoietic neoplasms. PMID- 8418382 TI - Blast colony-forming cells in myelodysplastic syndrome: decreased potential to generate erythroid precursors. AB - In vitro colony-forming abilities of highly purified primitive hematopoietic cells in eight cases of myelodysplastic syndrome (MDS) were studied using the blast cell colony assay. Blast cell colony formation from seven normal bone marrow (NBM) samples was studied in parallel. Blast cell colonies were formed in 7/8 cases of MDS, the numbers not significantly differing from those generated by NBM. In contrast the more mature hematopoietic progenitors (granulocyte-erythroid macrophage-megakaryocyte colony-forming unit, CFU-GEMM; erythroid burst-forming units, BFU-E; granulocyte colony-forming units, CFU-G; eosinophilic colony forming units, CFU-Eo) were severely depressed in numbers in MDS marrow. After replating of blast cell colonies in secondary cultures, colonies were obtained in 5/8 MDS cases. A marked difference was evident in the composition of the secondary colonies between MDS and normal marrow. Whereas secondary colonies derived from normal blast cell colonies consisted of about 45% of erythroid cells, MDS blast colonies generated mainly colonies of the granulocytic-monocytic lineage and no erythroid colonies. The normal quantitative level of CFU-blast progenitors in MDS in the context of their impaired ability to generate lineage specific progeny upon secondary plating suggests that the incompetence of maturation of MDS may reside in the CFU-blast progenitor cell being incapable of properly responding to growth factor stimulation. PMID- 8418383 TI - Sequential study of myeloid differentiation antigens of neutrophil granulocytes in different phases of chronic myeloid leukaemia: natural history and prognostic significance. AB - In an attempt to contribute to the knowledge of the natural history of Philadelphia-chromosome-positive chronic myeloid leukaemia (CML) and its prognosis, we analyzed sequentially the myeloid differentiation antigens of peripheral blood neutrophil granulocytes (NG) in different evolutive stages of the disease. Four monoclonal antibodies (CD15, CD24, 31D8, and 13F6) were used, and a total number of 116 sequential studies were performed in 43 patients. At diagnosis, there is a significant decrease of NG expressing myeloid differentiation antigens, which recover to nearly normal levels after initial control of the disease. The onset reduction is probably due to the circulation of incompletely mature NG. In accelerated/blastic phase NG expressing myeloid differentiation antigens decrease again, probably due to a true antigen loss. This reduction could herald by a few months the development of accelerated/blastic phase. In such a case, its predictive strength is higher than that of the well recognized initial prognostic parameters in CML. These results indicate that the sequential study of NG myeloid differentiation antigens may contribute to both a better understanding of the natural history of CML and the evolutive prognosis of this disease. PMID- 8418385 TI - Computer tools that crack the rules. PMID- 8418384 TI - Adhesion of precursor-B acute lymphoblastic leukaemia cells to bone marrow stromal proteins. AB - Adhesion to bone marrow stroma is a key event in normal B lymphopoiesis, allowing exposure of B-cell progenitors to regulatory cytokines. In order to investigate whether similar processes are important in the proliferation of acute lymphoblastic leukaemia (ALL) cells of precursor-B type, the expression of various adhesion molecules was examined. By flow cytometry analysis, CD-44 and the integrins VLA-4 and VLA-5 were the most prominent. CD-44 and VLA-4 were expressed on all 18 cases of precursor-B ALL analysed, while VLA-5 was found on 15 of 18 cases. The integrin CD-11a was detected on 8 of 11 cases, while its ligand, CD-54, was present in 6/12. Other adhesion proteins such as beta 3 integrin, CD-56, CD-15, and Leu8 were not expressed to any significant extent. In view of the known binding of VLA-4 and VLA-5 to extracellular fibronectin (FN), the adhesion of leukaemic cells to FN was evaluated in a colorimetric assay. The precursor-B ALL cell lines REH and KM-3, and 7/15 cases of precursor-B ALL, showed detectable binding to FN. Binding to the other extracellular matrix proteins collagen type 1 and vitronectin was not observed, although two ALL cases showed some binding to laminin. The functional activity of the VLA-4 and VLA-5 molecules was examined using an inhibitory peptide and monoclonal antibodies. These studies indicated that ALL cells adhere to soluble fibronectin predominantly through the VLA-5 molecule (blockable with the PHM-2 antibody and a peptide containing the RGD sequence) although binding mediated by VLA-4 was also apparent in some experiments (blockable by a 40 kDa fragment containing the heparin-binding domain of FN and inhibitory antibodies). These results indicate that precursor-B ALL cells may adhere to marrow stroma through interaction of VLA 4 and VLA-5 with FN, although other mechanisms of adhesion may be important. PMID- 8418386 TI - Development of an automated antibiotic consultant. AB - Physicians frequently need to order antibiotic therapy before the results of bacterial cultures and antibiotic susceptibility tests are available. Therapy selected empirically should be chosen on the basis of the most recent information about probable pathogens and their susceptibility to various drugs. We have developed a series of computer programs to help physicians identify the antibiotic regimens with the highest probability of acting against the pathogens that are most likely to be present at suspected sites of infection in individual patients. This automated antibiotic consultant is now available on our hospital information system. A comparison of the antibiotic regimens suggested by the computer with those indicated by cultures and susceptibility tests showed that the computer suggested an appropriate regimen for 717 of 761 culture events (94%). PMID- 8418387 TI - Implementation of COSTAR in an academic group practice of general pediatrics. AB - High-quality medical care requires a medical record that is complete, legible, and readily available. Information storage and retrieval are often more difficult in academic group practices than in private offices because of the complexity of the delivery system. We implemented a computerized medical record (COSTAR) in our academic group practice in pediatrics, and recorded data on 14,486 visits for preventive health care or illness over an 18-month period. Except when follow-up visits were made within 48 hours of an encounter, a complete medical record was available at all times. The attending staff physicians were enthusiastic about the computer-based record, and after a period of adaptation, the residents were as well. The nurses and clerical staff agreed that it improved office efficiency and thus patient care. Yet the system was abandoned in the face of medical college politics and problems in other practices at our institution. PMID- 8418388 TI - Permutation tests and "statistical significance." 1977. PMID- 8418389 TI - Fundamentals of image processing for personal computers. AB - Photographic images make an important contribution to medical education. Conventional means of display have included 35-mm projection, film, videotape, and more recently, analog videodiscs that accompany computer-based text. The availability of compact discs, with their enormous storage capacity, and of powerful central processing units now permits consideration of digital storage of visual images for display by the personal computer. The data processing required for the representation and transfer of high-quality images is forbidding, however, even with today's technology. The problem is compounded if one anticipates display of full-motion video. In this tutorial we first review the nature of analog signals and the process of capturing them in digital format. We then consider mass storage and the concept of compressing data into smaller amounts of information that can be processed and transferred by personal computers with the use of optical disc drives. A look into the future provides a vision of high-definition television-quality video in digital format. PMID- 8418390 TI - Computer-generated discharge instruction sheets. PMID- 8418391 TI - The problem with software reviews of decision support systems. PMID- 8418392 TI - Acute respiratory illness linked to use of aerosol leather conditioner--Oregon, December 1992. AB - At 8 a.m. on December 27, 1992, the Oregon Poison Center (OPC) notified the Oregon Health Division (OHD) that 13 persons in one household became ill following the use of an aerosol leather conditioner and that this report was similar to two reports received on December 26 that also involved use of this product. A review of telephone logs identified similar calls on December 23 and 24, for a total of 29 persons in six households who reported illness associated with use of this spray. By midday on December 27, the product producer issued a voluntary nationwide recall of this product. Following the public announcement of the recall, as of December 31, the number of preliminary reports to the OHD and the OPC of illness associated with use of this spray increased to 400 and involved approximately 550 persons. This report summarizes the preliminary findings of the ongoing investigation of this problem by the OHD. PMID- 8418393 TI - Air pollution information activities at state and local agencies--United States, 1992. AB - Because air pollution is a pervasive environmental health problem in the United States, one of the national health objectives for the year 2000 is to increase from 49.7% to 85.0% the proportion of persons who live in counties that have not exceeded any air quality standard during the previous 12 months (1). Public support for air pollution control efforts is critical if this national health objective is to be achieved. To characterize public health information activities related to air pollution, in 1992, the State and Territorial Air Pollution Program Administrators (STAPPA) and the Association of Local Air Pollution Control Officials (ALAPCO), with the assistance of CDC, conducted a survey of state and local air pollution control agencies. This report summarizes the findings of that survey. PMID- 8418394 TI - Deaths and hospitalizations from chronic liver disease and cirrhosis--United States, 1980-1989. AB - In 1989, chronic liver disease, including cirrhosis, was the ninth most frequent cause of death in the United States (1). Periodic analysis of trends and factors related to preventable death and hospitalization for chronic liver disease may be used to target prevention and control programs. This report examines national trends in death and hospitalization rates and state-specific death rates for chronic liver disease using data from CDC's National Center for Health Statistics' multiple-cause-of-death file and the National Hospital Discharge Survey (NHDS). PMID- 8418396 TI - Surveillance of the health status of Bhutanese refugees--Nepal, 1992. AB - From February 1991 through July 1992, 67,000 Bhutanese of Nepalese ethnic origin entered the Jhapa and Morang districts of southeastern Nepal (Figure 1) because of ethnic persecution in Bhutan. Six refugee camps were established along the Nepal-India border to accommodate the refugees. In July 1992, to assess the public health needs of these refugees, the Office of the United Nations High Commissioner for Refugees (UNHCR), the Save the Children Fund (SCF), and CDC established a surveillance system to monitor morbidity and mortality. This report describes the surveillance system implemented in these six camps in July 1992 and presents mortality data collected from March through July 1992. PMID- 8418395 TI - Hepatitis E among U.S. travelers, 1989-1992. AB - Outbreaks of hepatitis E (i.e., enterically transmitted non-A, non-B hepatitis) have occurred in some parts of the world and have generally been related to contaminated water supplies. Until recently, when research-based serologic tests (1,2) were developed to test for antibody to hepatitis E virus (anti-HEV), no serologic test was available to identify HEV infection, and diagnosis depended on a history of exposure in an appropriate epidemiologic setting and the exclusion of other causes of viral hepatitis. During 1989-1992, acute HEV infection was documented among six persons in the United States who had returned from international travel. This report summarizes CDC's serologic documentation of acute HEV infection--presumed to have been acquired during international travel- in four of these persons. PMID- 8418397 TI - Projections of the number of persons diagnosed with AIDS and the number of immunosuppressed HIV-infected persons--United States, 1992-1994. PMID- 8418398 TI - Surveillance of deaths attributed to a Nor'easter--December 1992. AB - During December 10-13, 1992, a severe weather system of snow, sleet, rain, and high winds struck Connecticut, Delaware, Maryland, Massachusetts, New Jersey, New York, Pennsylvania, Rhode Island, Virginia, and West Virginia. The highest recorded winds from this winter storm, called a nor'easter, were 80 miles per hour (mph) gusts at Cape May, New Jersey, with sustained winds of 20-30 mph. The tidal surge was 1-4 feet above normal, and wave heights were 20-25 feet near the shore. The 24-hour snowfall was 27 inches in the hills west of Boston. Flooding was recorded at 4-5 feet in both Boston and New York City. In the Berkshire Mountains in western Massachusetts, 4 feet of snow fell, with drifts as high as 10 feet. This report summarizes findings of surveillance for deaths associated with this storm and is based on information obtained from medical examiner (ME) offices. PMID- 8418399 TI - Respiratory syncytial virus outbreak activity--United States, 1992. AB - The National Respiratory and Enteric Virus Surveillance System (NREVSS) was established in 1989 to monitor trends in respiratory syncytial virus (RSV), parainfluenza viruses, adenoviruses, and rotaviruses in the United States and to provide information to public health officials and health-care providers about the presence of these viruses in their communities. Based on reports of RSV detections to the NREVSS, during the 1992-93 season, outbreaks of RSV had occurred in all regions of the United States by December 1992. This report summarizes surveillance results for RSV-antigen detections from June 27, 1992, through December 12, 1992, and assesses trends in RSV from July 1, 1990, through December 12, 1992. PMID- 8418400 TI - Condom use and sexual identity among men who have sex with men--Dallas, 1991. AB - Safer sex practices intended to reduce the risk for human immunodeficiency virus (HIV) infection have been vigorously promoted among men who are homosexual or bisexual (i.e., men who have sex with men). Such efforts have emphasized personal responsibility and protection of partners, and many of these men appear to have adopted risk-reducing behaviors (1). However, it is unknown whether these safer sex practices and norms have been adopted by men who have sex with men but conceal their sexual orientations or do not self-identify as homosexual or bisexual (2-7). To characterize the relation between the adoption of safer sex practices among men who have sex with men and sexual self-identity, as well as HIV information-seeking, exposure to the homosexual or bisexual community culture, and comfort in disclosing sexual identity, the Dallas County (Texas) Health Department (DCHD) conducted a survey among men who have sex with men. This report summarizes survey findings for men who reported having had sex with men and who visited DCHD clinics for anonymous HIV counseling and testing from January through June 1991. PMID- 8418401 TI - Efficacy of carotid endarterectomy for asymptomatic carotid stenosis. The Veterans Affairs Cooperative Study Group. AB - BACKGROUND: The efficacy of carotid endarterectomy in patients with asymptomatic carotid stenosis has not been confirmed in randomized clinical trials, despite the widespread use of operative intervention in such patients. METHODS: We conducted a multicenter clinical trial at 11 Veterans Affairs medical centers to determine the effect of carotid endarterectomy on the combined incidence of transient ischemic attack, transient monocular blindness, and stroke. We studied 444 men with asymptomatic carotid stenosis shown arteriographically to reduce the diameter of the arterial lumen by 50 percent or more. The patients were randomly assigned to optimal medical treatment including antiplatelet medication (aspirin) plus carotid endarterectomy (the surgical group; 211 patients) or optimal medical treatment alone (the medical group; 233 patients). All the patients at each center were followed independently by a vascular surgeon and a neurologist for a mean of 47.9 months. RESULTS: The combined incidence of ipsilateral neurologic events was 8.0 percent in the surgical group and 20.6 percent in the medical group (P < 0.001), giving a relative risk (for the surgical group vs. the medical group) of 0.38 (95 percent confidence interval, 0.22 to 0.67). The incidence of ipsilateral stroke alone was 4.7 percent in the surgical group and 9.4 percent in the medical group. An analysis of stroke and death combined within the first 30 postoperative days showed no significant differences. Nor were there significant differences between groups in an analysis of all strokes and deaths (surgical, 41.2 percent; medical, 44.2 percent; relative risk, 0.92; 95 percent confidence interval, 0.69 to 1.22). Overall mortality, including postoperative deaths, was primarily due to coronary atherosclerosis. CONCLUSIONS: Carotid endarterectomy reduced the overall incidence of ipsilateral neurologic events in a selected group of male patients with asymptomatic carotid stenosis. We did not find a significant influence of carotid endarterectomy on the combined incidence of stroke and death, but because of the size of our sample, a modest effect could not be excluded. PMID- 8418402 TI - Early treatment of acute biliary pancreatitis by endoscopic papillotomy. AB - BACKGROUND: Most patients with acute biliary pancreatitis have stones in the biliary tract or ampulla of Vater. Because these stones may be passed spontaneously soon after a patient is admitted to the hospital, the importance of early operative removal is not known. We tested the hypothesis that endoscopic papillotomy within 24 hours of admission decreased the incidence of complications in patients with acute biliary pancreatitis. METHODS: We studied 195 patients with acute pancreatitis who were randomly assigned to one of two groups: 97 patients underwent within 24 hours after admission emergency endoscopic retrograde cholangiopancreatography (ERCP) followed by endoscopic papillotomy for ampullary and common-bile-duct stones, and 98 patients received initial conservative treatment and selective ERCP with or without endoscopic papillotomy only if their condition deteriorated. RESULTS: One hundred twenty-seven patients ultimately proved to have biliary stones. Emergency ERCP with or without endoscopic papillotomy resulted in a reduction in biliary sepsis as compared with conservative treatment (0 of 97 patients vs. 12 of 98 patients, P = 0.001). The decrease in biliary sepsis occurred both in patients predicted to have mild pancreatitis (0 of 56 patients in the group that received emergency ERCP vs. 4 of 58 patients in the conservative-treatment group, P = 0.14) and in patients predicted to have severe pancreatitis (0 of 41 patients vs. 8 of 40 patients, P = 0.008). In all patients who had unrelenting biliary sepsis, persistent ampullary or common-bile-duct stones were identified. There were no major differences in the incidence of local complications (10 patients in the group that received emergency ERCP vs. 12 patients in the conservative-treatment group) or systemic complications (10 patients vs. 14 patients) of acute pancreatitis between the two groups, but the hospital mortality rate was slightly lower in the group undergoing emergency ERCP with or without endoscopic papillotomy (5 patients vs. 9 patients, P = 0.4). CONCLUSIONS: Emergency ERCP with or without endoscopic papillotomy is indicated in the treatment of patients with acute pancreatitis. PMID- 8418403 TI - A prospective, randomized study of cochlear implants. The Department of Veterans Affairs Cochlear Implant Study Group. AB - BACKGROUND: Cochlear implants restore some degree of hearing in patients with severe hearing impairment, but the efficacy of different implants has not been compared. We conducted a prospective trial to compare several cochlear implants. METHODS: We studied 82 patients who were randomly assigned to receive one of three cochlear implants: the Ineraid multichannel implant (implant 1), the Nucleus multichannel implant (implant 2), and the 3M/Vienna single-channel implant (implant 3). All the patients had profound deafness, and none had derived benefit from hearing aids. The assigned device was successfully implanted in 80 patients. Twenty-four hearing tests were used to assess the patients' performance before implantation and 12 and 24 months after implantation. The tests were grouped into five categories according to their content, and a weighted composite index was developed to provide a single numerical indicator of the overall auditory response. RESULTS: All the patients were able to hear with their implants. Nineteen of the 30 patients (63 percent) who received implant 2, 18 of the 30 patients (60 percent) who received implant 1, and 1 of the 20 patients (5 percent) who received implant 3 were able to distinguish some words and sentences. The scores for the composite index were similar in the patients who received implant 1 and those who received implant 2, and were higher in both these groups than in the patients who received implant 3 (P = 0.02). When 24 patients with implant 2 were given an improved speech processor, their composite index increased significantly within 3 months (P < 0.001); their score at that time was also significantly higher (P = 0.04) than the score of the patients with implant 1 at 24 months. Age at implantation, lip-reading ability, and IQ were prognostic indicators of the patients' performance with a cochlear implant. CONCLUSIONS: Multichannel cochlear implants are superior to single-channel implants, especially for understanding speech. Changes in speech processing can improve patients' performance. PMID- 8418404 TI - The relation between insulin sensitivity and the fatty-acid composition of skeletal-muscle phospholipids. AB - BACKGROUND: Insulin resistance and hyperinsulinemia are features of obesity, non insulin-dependent diabetes mellitus, and other disorders. Skeletal muscle is a major site of insulin action, and insulin sensitivity may be related to the fatty acid composition of the phospholipids within the muscle membranes involved in the action of insulin. METHODS: We determined the relation between the fatty-acid composition of skeletal-muscle phospholipids and insulin sensitivity in two groups of subjects. In one study, we obtained samples of the rectus abdominis muscle from 27 patients undergoing coronary artery surgery; fasting serum insulin levels provided an index of insulin sensitivity. In the second study, a biopsy of the vastus lateralis muscle was performed in 13 normal men, and insulin sensitivity was assessed by euglycemic-clamp studies. RESULTS: In the patients undergoing surgery, the fasting serum insulin concentration (a measure of insulin resistance) was negatively correlated with the percentage of individual long chain polyunsaturated fatty acids in the phospholipid fraction of muscle, particularly arachidonic acid (r = -0.63, P < 0.001); the total percentage of C20 22 polyunsaturated fatty acids (r = -0.68, P < 0.001); the average degree of fatty-acid unsaturation (r = -0.61, P < 0.001); and the ratio of the percentage of C20:4 n-6 fatty acids to the percentage of C20:3 n-6 fatty acids (r = -0.55, P < 0.01), an index of fatty-acid desaturase activity. In the normal men, insulin sensitivity was positively correlated with the percentage of arachidonic acid in muscle (r = 0.76, P < 0.01), the total percentage of C20-22 polyunsaturated fatty acids (r = 0.76, P < 0.01), the average degree of fatty-acid unsaturation (r = 0.62, P < 0.05), and the ratio of C20:4 n-6 to C20:3 n-6 (rho = 0.76, P = 0.007). CONCLUSIONS: Decreased insulin sensitivity is associated with decreased concentrations of polyunsaturated fatty acids in skeletal-muscle phospholipids, raising the possibility that changes in the fatty-acid composition of muscles modulate the action of insulin. PMID- 8418405 TI - Unconventional medicine in the United States. Prevalence, costs, and patterns of use. AB - BACKGROUND: Many people use unconventional therapies for health problems, but the extent of this use and the costs are not known. We conducted a national survey to determine the prevalence, costs, and patterns of use of unconventional therapies, such as acupuncture and chiropractic. METHODS: We limited the therapies studied to 16 commonly used interventions neither taught widely in U.S. medical schools nor generally available in U.S. hospitals. We completed telephone interviews with 1539 adults (response rate, 67 percent) in a national sample of adults 18 years of age or older in 1990. We asked respondents to report any serious or bothersome medical conditions and details of their use of conventional medical services; we then inquired about their use of unconventional therapy. RESULTS: One in three respondents (34 percent) reported using at least one unconventional therapy in the past year, and a third of these saw providers for unconventional therapy. The latter group had made an average of 19 visits to such providers during the preceding year, with an average charge per visit of $27.60. The frequency of use of unconventional therapy varied somewhat among socio-demographic groups, with the highest use reported by nonblack persons from 25 to 49 years of age who had relatively more education and higher incomes. The majority used unconventional therapy for chronic, as opposed to life-threatening, medical conditions. Among those who used unconventional therapy for serious medical conditions, the vast majority (83 percent) also sought treatment for the same condition from a medical doctor; however, 72 percent of the respondents who used unconventional therapy did not inform their medical doctor that they had done so. Extrapolation to the U.S. population suggests that in 1990 Americans made an estimated 425 million visits to providers of unconventional therapy. This number exceeds the number of visits to all U.S. primary care physicians (388 million). Expenditures associated with use of unconventional therapy in 1990 amounted to approximately $13.7 billion, three quarters of which ($10.3 billion) was paid out of pocket. This figure is comparable to the $12.8 billion spent out of pocket annually for all hospitalizations in the United States. CONCLUSIONS: The frequency of use of unconventional therapy in the United States is far higher than previously reported. Medical doctors should ask about their patients' use of unconventional therapy whenever they obtain a medical history. PMID- 8418406 TI - Drowning. PMID- 8418407 TI - New concepts about the mast cell. PMID- 8418408 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 4-1993. A 73-year-old man with severe facial pain, visual loss, decreased ocular motility, and an orbital mass. PMID- 8418409 TI - Carotid endarterectomy for asymptomatic carotid stenosis. PMID- 8418410 TI - Endoscopic sphincterotomy in the early treatment of acute pancreatitis. PMID- 8418411 TI - A brief perspective on cochlear implants. PMID- 8418412 TI - Why unconventional medicine? PMID- 8418413 TI - Nitrous oxide and fertility. PMID- 8418415 TI - Pounding in the neck in supraventricular tachycardia. PMID- 8418414 TI - The racial disparity in very low birth weight. PMID- 8418416 TI - Pounding in the neck in supraventricular tachycardia. PMID- 8418418 TI - Effect of L-arginine on plasminogen-activator inhibitor in hypertensive patients with hypercholesterolemia. PMID- 8418417 TI - Pounding in the neck in supraventricular tachycardia. PMID- 8418419 TI - Does a clean-catch urine sample reduce bacterial contamination? PMID- 8418421 TI - Measuring professional nursing practice. PMID- 8418420 TI - Clinical problem-solving: interpreting hoofbeats: can Bayes help clear the haze? PMID- 8418422 TI - Advanced practice: the National Council responds. PMID- 8418423 TI - Legal update for nurses--1992, Part I. PMID- 8418424 TI - Keeping your PC hardware healthy. PMID- 8418425 TI - A lethal attitude. PMID- 8418426 TI - Getting ready for the new tomorrow. PMID- 8418427 TI - A call for reform of our nursing education system. American Organization of Nurse Executives. PMID- 8418428 TI - Total quality management: doing things right. AB - Total quality management transforms quality assurance performance controls into a dynamic system for continuously improving healthcare services to internal and external "customers" in terms of outcomes. Success depends upon gaining and maintaining commitment of physicians, top management and key staff members through a working environment which values and rewards improvement efforts. PMID- 8418429 TI - Continuous quality improvement in shared leadership. PMID- 8418430 TI - Quality indicators: control maintains--propriety improves. AB - Continuous improvement of care depends upon the efforts of all nursing personnel. Supervisors facilitate quality program operations in their specific areas; head nurses document outcomes; and staff nurses identify problems and contribute to planning. All join in evaluating appropriateness of patient care and in modifying approaches for greater effectiveness. PMID- 8418431 TI - The goal is quality improvement. PMID- 8418432 TI - Work role inventory: a guide to job satisfaction. AB - The Miller-Carey Work Role Expectations model helps the nurse executive understand the dynamics which motivate managerial teams. The application of this model emphasizes the importance of assessing nurse manager's functions and responsibilities and suggests specific ways to enhance work motivation. PMID- 8418433 TI - Clerical activities and the nurse manager. AB - A nurse manager identifies, records and reassigns clerical activities on her two Pediatric Special Care Units. This study shows nurse managers how to reduce their clerical time and free managerial time for improving unit operations. PMID- 8418434 TI - Managing job satisfaction for the long run. AB - Twenty senior nurse executives were surveyed to determine their perceptions of job satisfaction and reward systems among long-term, mid-career employees. These needs go beyond basic security and into higher levels of professionalism. PMID- 8418436 TI - Health: reform, refrain or reshuffle? PMID- 8418435 TI - The nurse executive in Operation Desert Shield/Desert Storm. AB - The Army Medical Department's Chief Nurse describes a corporate nurse executive's role in the Persian Gulf campaign to free Kuwait in early 1991. Although language and cultural barriers presented problems, U.S. Army and Host Nation nurses were able to integrate their resources to assure smooth operations during wartime. PMID- 8418437 TI - Home care: lest we forget house calls. PMID- 8418438 TI - Retention: owning the issue. PMID- 8418440 TI - Gowns are "protective"--not barriers! PMID- 8418439 TI - Answering the "phone:" risks and rewards. PMID- 8418441 TI - Moving in a new direction: computerized adaptive testing (CAT). PMID- 8418443 TI - [New cases of celiac disease detected by anti-endomysial antibody test in families of gluten-sensitive patients and among children examined for non specific gastrointestinal complaints]. AB - Among 228 relatives of 101 gluten-sensitive patients, 13 anti-endomysium antibody (EmA) positive persons (7 children and 6 adults) were identified. In 12/13 cases jejunal biopsy confirmed severe villous atrophy consistent with celiac disease. In the single EmA positive sibling without villous atrophy the histology is thought to be influenced by a steroid treatment because of pulmonary disease. By routine EmA-testing 12 unexpected EmA positive patients were found out of 756 children with complaints and laboratory results otherwise not justifying jejunal biopsy at the first evaluation. Their initial diagnoses were: proteinuria, colitis, Crohn's disease, rickets, recurrent vomiting, resolved postinfectious lactase deficiency, "previously excluded" celiac disease. Severe villous atrophy could be demonstrated in all EmA positive patients subsequently. In further 204 EmA negative children the biopsy showed no atrophy. EmA positivity may reveal clinically not apparent severe villous atrophy emphasizing the role of a new non invasive and highly specific serological screening method for celiac disease. PMID- 8418442 TI - [Echocardiographic detection and follow up of left ventricular thrombosis following myocardial infarct]. AB - 914 patients with myocardial infarction have been examined during the last 3 years. Left ventricular thrombi were found in 74 patients (8%), (58 male and 16 female, median age 57.3 years). Thrombi were found in 37 cases, oval protruding in 28 cases, pedicled or multiple in 9 cases. Thrombus was most frequently located to the apex (61 cases), while in 8 patients localization was anteroseptal, in 4 patients anterolateral and in 1 patient inferior. 36 patients had also ventricular aneurysm of which 23 were at the apex and 13 at the anterior wall. After anticoagulant therapy the patients were controlled by echocardiography. In 23 out of the 53 patients that were restudied the thrombus had resolved, in 8 patients thrombus has decreased in size and in 22 patients there was no change in thrombus size. Embolism has occurred in 13 cases of which 10 were cerebral embolism. During the follow-up period (mean 12.8 months) six patients died. On the basis of their study the authors conclude that thrombosis is a frequent complication of anterior myocardial infarction and may lead to embolic events, so anticoagulant therapy controlled by echocardiography is needed, because of it's efficacy in the resolution of thrombus and eliminating the risk of embolism. PMID- 8418444 TI - [The importance of recognizing oligohydramnios]. AB - Outcome of 150 pregnancies complicated with oligohydramnios were examined. The results were compared to healthy pregnancies. This was the control group. It has been found that occurrence of the intrauterine growth retardation and signs of the intrauterine distress in the pathological group are more frequent than in the control group. After comparing the body length and the body weight of the newborns we found a significant difference between the two groups. The authors emphasize the significance of the recognition in time of oligohydramnios and the importance of ultrasound examination during pregnancy. PMID- 8418445 TI - [Bronchoalveolar lavage in the confirmation of pulmonary involvement in chronic lymphoid leukemia]. AB - The authors made bronchoalveolar lavage (BAL) for explaining the origin of diffuse bilateral lung--infiltration in patients with chronic lymphoid leukaemia. On the base of lavage-fluid, the lung damage may be caused by leukaemic infiltration. The lesion has not changed by antibiotic therapy, but was radiologically cured after giving corticoids. The authors have not read about approaching chronic lymphoid leukaemic pulmonary infiltration by BAL. They call attention to the etiologic clarification of lung organs' complications in haematological aspects and outstanding role of BAL as a non-invasive method. PMID- 8418447 TI - [Analysis of a one-year caseload of a pediatric health center in the community of Barcs with special reference to obstructive bronchitis]. PMID- 8418446 TI - [Complications of Crohn disease, surgical indications, surgical solutions and recurrence]. AB - There is not another disease, of which complications would be such frequent and various as of Crohn's disease. One of the most common complications is intestinal obstruction, caused by angulation and stricture, as a result of inflammatory granulomas. Fistulas between affected intestines and cavital organs or the abdominal wall can also often be observed. The likelihood of appearing complications is correlating with the time from the onset of the disease. If they are dangerous for the patient's life, surgical intervention is required. In some of the cases, when complications don't threaten the patient's life, medical treatment may be beneficial. Surgery is indicated only in those cases, when complications mean danger, or the continuous active medical treatment is insufficient. Most frequently the resection of the damaged intestine is performed, nowadays stricture plasty is also recommended. The operative mortality is 4.1%. The disease is featured by frequent recurrences: 28% to 33.3%. The current operative mortality is 18.6%. The prognosis is always uncertain, because recurrence may occur many years after operation. PMID- 8418448 TI - Bacterial meningitis. PMID- 8418449 TI - Nephrotic syndrome. PMID- 8418450 TI - School readiness: an overview for pediatricians. PMID- 8418451 TI - Serious complications of acute sinusitis. AB - Acute sinusitis is common and usually the result of edema around the sinus ostia from the common cold. Chronic sinusitis and recurrent sinusitis are common complications of untreated or undertreated infection. Serious and life threatening complications are uncommon but demand a high index of suspicion, accurate diagnosis, and rapid intervention by medical and surgical methods. Immediate parenteral administration of antibiotics may be necessary to prevent permanent disabilities or death, and early consultation with an otolaryngologist or neurosurgeon is prudent. Computed tomography is the imaging method of choice for detection of serious intracranial complications of acute sinusitis. Increasing numbers of immunocompromised patients and seriously ill hospitalized patients with problems related to complications of acute sinusitis are being seen. PMID- 8418453 TI - All in the family. PMID- 8418452 TI - Current perspectives on respiratory syncytial virus infection. AB - Respiratory syncytial virus (RSV) is the most important cause of acute viral bronchiolitis and pneumonia in infants and children. Management of RSV infection has changed dramatically over the past 5 to 7 years because of (1) the advent of widely available rapid tests for its specific identification and (2) use of the antiviral agent ribavirin (Virazole). Although RSV infection remains a life threatening disease, particularly in certain high-risk groups, appropriate management and judicious use of ribavirin should help physicians reduce its morbidity and mortality. PMID- 8418454 TI - Pulmonary manifestations of rheumatic disease. AB - The lungs are a common target in many rheumatic diseases. Treatment of the primary rheumatic disease is often all that is required to control lung involvement. Physicians must be careful not to attribute lung disease to the underlying rheumatic disorder, because interstitial lung disease can be a complication of drug therapy. PMID- 8418455 TI - Ascariasis. An infection to watch for in immigrants. AB - Infection with Ascaris lumbricoides, a roundworm that invades the human digestive tract, affects about 25% of the world's population. Manifestations of ascariasis vary and include constitutional symptoms, particularly pulmonary and gastrointestinal complaints. Complications include pneumonitis, intestinal obstruction, and damage to vital organs. Diagnosis is made in most cases by sampling stool for ova and parasites. Treatment with an anthelmintic agent is usually effective in mild cases, and prognosis is excellent. More severe infection may cause significant morbidity and require surgical intervention. PMID- 8418456 TI - Current management of symptomatic gallstones. AB - Present medical alternatives for the treatment of symptomatic cholelithiasis have been shown to be safe but of limited efficacy. The only medical option approved by the Food and Drug Administration is dissolution with an oral agent. This therapy is appropriate for patients with mild or infrequent symptoms who are poor operative risks or have a low risk of recurrence (ie, long-term users of nonsteroidal anti-inflammatory agents or those who have single stones). Now and for the foreseeable future, laparoscopic cholecystectomy is the procedure of choice for treatment of symptomatic cholelithiasis. PMID- 8418457 TI - Noncontraceptive benefits of oral contraceptive agents. Patients should know the positive effects. AB - Physicians' role as teachers and counselors includes the responsibility to provide detailed and accurate information on the risks and benefits of therapy whenever a prescription is written. Inaccurate or overly negative communication, particularly from the mass media, may confuse patients trying to make an informed decision regarding use of oral contraceptives. Patients have the right to be informed of possible prevention of life-threatening disease and improvement of quality of life as a result of oral contraceptive therapy. PMID- 8418458 TI - Wegener's granulomatosis. Typical symptoms in an atypical patient. AB - Wegener's granulomatosis classically involves the respiratory tract and kidneys, and the disorder may be confused with any of a number of diseases having similar symptoms. Although most common in middle-aged whites, Wegener's granulomatosis may occur in atypical patients, as in the case described here. Accurate diagnosis depends on lung biopsy, and prompt treatment prevents permanent damage and significant sequelae. PMID- 8418459 TI - Thyroid hormone therapy. What, when, and how much. AB - Hypothyroidism is the condition most commonly treated with exogenous thyroid hormone. The goal of therapy is to normalize levels of serum thyrotropin (thyroid stimulating hormone), which should be monitored by a high-sensitivity test. Adjustments in optimal dose may be necessary for a number of physiologic reasons (eg, decreased gastrointestinal absorption, pregnancy). Thyroid hormone therapy is also appropriate after surgery for thyroid cancer and for patients with goiter or benign thyroid nodules. In the absence of hypothyroidism, such treatment should not be used for obesity, fatigue, irregular menses, or infertility. PMID- 8418460 TI - A look at Clinton's new health team. PMID- 8418461 TI - Insomnia. Use of a 'decision tree' to assess and treat. AB - Insomnia, a remarkably common disturbance in a basic biologic function, arises from multiple psychological, physiologic, and environmental factors. Transient insomnia usually resolves spontaneously. Short-term insomnia is usually normalized by coping with acute changes in a medical condition or a relationship. In patients with insomnia caused by stressful life events, a short (ie, 10 days or less) course of a short- or intermediate-acting benzodiazepine hypnotic may be indicated. Long-term insomnia deserves comprehensive evaluation. Psychiatric disorders are common in patients with long-term insomnia. In patients over age 50, intrinsic sleep disorders are more prevalent. Behavioral therapy, including improved sleep hygiene, stimulus-control techniques, and sleep-restriction therapy, is preferred in the management of long-term insomnia. Pharmacotherapy (eg, low-dose antidepressant or benzodiazepine) is best used as an adjunct. PMID- 8418462 TI - Osteoarthritis treatment update. Minimizing pain while limiting patient risk. AB - Osteoarthritis is a chronic disease that has exacerbations and remissions. Pain is the symptom that patients want addressed. It is important to remember, however, that simple alleviation of pain does not alter the underlying problem. Attention must be directed toward using physical therapy and other physical measures in conjunction with pharmacologic intervention for symptom relief. Use of simple analgesic agents is the safest initial approach, perhaps in conjunction with topical treatment with a compound such as capsaicin (Zostrix). If pain relief is inadequate, use of nonsteroidal anti-inflammatory agents should be considered, with careful monitoring of gastrointestinal symptoms and renal status, particularly in the elderly. For flares of disease, intra-articular injection of a corticosteroid may give short-term relief. Relief of pain and restoration of function can be accomplished in some patients with early disease, particularly if an integrated approach to treatment is used. Advanced disease can be made more tolerable but may eventually require surgical intervention, which generally provides excellent results. PMID- 8418463 TI - Adult meningitis. Rapid identification for prompt treatment. AB - To be effective, treatment of meningitis should be based on the history and physical examination, careful examination of the cerebrospinal fluid, and good clinical judgment regarding the most likely pathogen. Meningitis in adults is usually caused by certain common viruses and bacteria, although atypical pathogens should be considered in immunocompromised patients. Supportive therapy measures are appropriate for viral disease, and intravenous acyclovir (Zovirax) may be given if infection with herpes simplex virus is suspected. In cases of presumed bacterial meningitis, antimicrobial agents should be selected that penetrate the blood-brain barrier and maintain activity against the most likely pathogens; antibiotic therapy should be instituted right away, along with supportive measures. Although corticosteroids have proven benefits in the treatment of pediatric populations with Haemophilus influenzae meningitis, their effectiveness in adults has not yet been established. Prophylaxis with vaccines or rifampin is sometimes useful. PMID- 8418464 TI - Variation in late potentials and the reproducibility of their measurement. AB - The usefulness of a test depends on its reproducibility. This determines how closely the test result indicates the actual pathophysiologic state, how well it will predict that state in the future, and if interventions or further pathologic changes are reflected by the test. There is a variation in the parameters of the signal-averaged ECG, more so with spectral than with time domain measurements. These must be accounted for when estimating risk. If one presumes that risk is proportional to the extent of abnormality, then the variation in measurements simply means that only borderline cases can potentially be miscategorized. More important, the lack of reproducibility of measurements made from the signal averaged ECG indicates that changes noted in an individual after an intervention, such as a surgical intervention, must be viewed with a jaundiced eye. Group changes are perhaps meaningful, and indicate a physiologic effect, but clinical decisions cannot be made unless the changes observed in an individual patient exceed the confidence limits of expected variation. There has been debate as to the usefulness of measurements made from the signal-averaged ECG in predicting antiarrhythmic drug effects (the effect of drugs is discussed elsewhere in this symposium). Here an analogy must be made to the suppression of asymptomatic ventricular ectopy. First, we cannot make a statement that there has been a drug effect unless the parameter measured changes beyond the confidence limits of normal variation or reproducibility. Second, we cannot translate a change in a measurement into a change in risk for arrhythmic events without subjecting that hypothesized relationship to a long-term placebo-controlled clinical trial, albeit acute electrophysiologic trials correlating changes in the signal-averaged ECG to ventricular tachycardia induction provide some insight. And perhaps the relationship must be tested independently for each drug assessed. In the same regard, there is much excitement about the benefits of thrombolytic therapy, but when diagnosing benefit to the individual patient we have to remember the lack of reproducibility of the measurements and also keep in mind that an improved signal averaged ECG cannot be translated into an improved prognosis without long-term controlled studies. In summarizing the variation and reproducibility of measurements made from the signal-averaged ECG we avoided providing more than a sense of the extent of variation expected because precise confidence intervals depend on the particular techniques used to make the measurements.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418465 TI - Post myocardial infarction stratification and the signal-averaged electrocardiogram. PMID- 8418466 TI - Dissociation of autoaggression and self-superantigen reactivity. AB - Self-superantigens have been described as products of endogenous retroviruses of the mouse ('minor lymphocyte stimulating loci') that are capable of interacting without prior processing with conserved domains of TCR V beta chains, causing the activation and deletion of most T cells expressing products of determined V beta gene families [1-4]. The fact that superantigens activate a far higher percentage of T cells (1-20%) than conventional, peptidic antigens (< 0.1%) provides the methodological advantage that the degree of clonal deletion may be measured by the analysis of the TCR repertoire using appropriate anti-V beta antibodies. Although much information on the spatio-temporal organization of repertoire purging has been gathered by virtue of self-superantigens, serious doubts exist as to the possibility that such structures serve as pathogenetically relevant autoantigens. Thus, certain inbred mice spontaneously develop autoimmune diseases, although they bear T-cell repertoires that appear to be purged from self-superantigen-reactive V beta products. In addition, therapeutic interventions targeted to V beta gene products that are not specific for self superantigens are successful in preventing disease development. The lack of correlation between superantigen-related V beta deletions and autoimmune disease development is substantiated in further models of murine autoimmunity. Based on these observations, we formulate the hypothesis that self-superantigen-reactive T cells are not involved in the development of autoimmune diseases. PMID- 8418467 TI - Idiotypic vaccination: immunoprotection mediated by anti-idiotypic antibodies with antibiotic activity. AB - Anti-Id antibodies were raised in mice against a monoclonal antibody (MoAb KT4) that neutralized the in vitro activity of a Pichia anomala yeast killer toxin. Monoclonal antibody was administered to BALB/C syngeneic mice with different schedules of immunization before intravenous challenge with increasing amounts of yeast killer toxin-sensitive Candida albicans cells. The course of candidosis was studied in comparison with mice non-immunized and immunized with an isotype matched unrelated MoAb subdivided into control groups. Protection was reflected by statistically significant increases in survival rate of mice immunized with MoAb KT4 which showed variable serum levels of yeast killer toxin-like anti-Id antibodies. MoAb KT4 affinity chromatography purified mouse anti-Id antibodies were capable of killing in vitro the yeast cells of the Candida albicans strain used for the experimental infection. This is the first report of antimicrobial protection that exploits the role of anti-idiotypic antibodies presumably acting in vivo as antibiotics (idiotypic vaccination). PMID- 8418468 TI - The secretion of the third component of complement (C3) by human polymorphonuclear leucocytes from both normal and systemic lupus erythematosus cases. AB - Recently, murine peritoneal exudate polymorphonuclear leucocytes (PMNs) have been proved to secrete complement C3. In this report we show the secretion of C3 by normal human blood PMNs. ELISA assay was used to detect secreted C3 in culture supernatants of PMNs, while immunoperoxidase staining was used for intracellular C3 detection. 12-o-tetradecanoyl phorbol 13 acetate (TPA) had a flushing effect on C3 secretion by PMNs but not macrophages, suggesting a special C3 storing capability in PMNs. Dioctanoyl glycerol, mezerein and calcium ionophore A23187 caused the same marked increase in C3 secretion by PMNs. This suggests the contribution of protein kinase C and the calmodulin pathway in the mechanism of C3 secretion, similar to murine peritoneal exudate PMNs. In some cases of systemic lupus erythematosus, C3 secretion by blood PMNs was increased but no similar response to TPA could be detected. PMID- 8418469 TI - Recombinant human tumour necrosis factor-alpha (rhTNF-alpha) and rhTNF-alpha analogue enhance amyloid deposition in the Syrian hamster. AB - Tumour necrosis factor-alpha (TNF-alpha) is one of the cytokines that stimulate the production of serum amyloid A (SAA), the precursor of AA amyloid. The role of TNF-alpha in amyloidogenesis was investigated in experimental hamsters using purified recombinant human TNF-alpha (rhTNF-alpha) and rhTNF-alpha analogue different from the normal molecule by two amino acid substitutions. Daily injections of 1 microgram rhTNF-alpha resulted in elevated SAA levels but even in the presence of amyloid enhancing factor (AEF) no amyloid was deposited, indicating that apart from the AEF and one particular SAA stimulating factor an additional factor is needed to result in amyloid deposition. This factor is generated by repeated injections of E. coli lipopolysaccharide (LPS). A single intraperitoneal injection of 12.5 micrograms or more of rhTNF-alpha followed by seven daily subcutaneous injections of LPS resulted in enhanced amyloid deposition. Heat denaturation of rhTNF-alpha did abolish its AEF activity. The rhTNF-alpha analogue, having one-fifth of the cytotoxic activity of the normal rhTNF-alpha, showed a similar reduction in its SAA-inducing capacity and its amyloidogenicity. This suggests the AEF activity to be closely related to TNF alpha activity. However, poly(I)-poly(C) (a potent inducer of IL-6) also showed AEF activity, suggesting that not a single cytokine but rather a certain combination of different cytokines could be decisive in AA amyloidogenesis. PMID- 8418471 TI - Tetranectin: a novel secretory protein from human monocytes. AB - Tetranectin is a recently described human plasma protein, which is found in most secretory cells throughout the body, including neutrophils. We present evidence for the presence of tetranectin in human monocytes and macrophages as well, and that these cells upon adherence or weak stimulation release a 20 kDa protein identified as tetranectin by immunoblotting. The amount of tetranectin released is 3-18 ng/h/10(6) monocytes. The possible influence of tetranectin on cellular functions was tested in migration and oxidative metabolism assays. Monocyte spontaneous migration was significantly stimulated by preincubation with purified tetranectin, whereas chemotactic and chemiluminescence responses to fMLP and C5a were unchanged. Neutrophil functions were not affected. It is concluded that tetranectin is secreted from human mononuclear phagocytes upon weak stimulation, and that the secreted tetranectin facilities spontaneous migration of these cells. PMID- 8418470 TI - Oligonucleotide probes to the 5' end of the framework 3 (FR3) gene segment detect polymorphisms of VH gene sequences encoding biologically important amino acid residues. AB - In this study, we demonstrate the use of oligonucleotide probes to the most VH family-specific framework, the Framework 3 (FR3) gene segment, in the analysis of germline DNA. Compared with the hybridization of restriction enzyme-digested genomic DNA to large, cloned human VH family probes, hybridization with oligonucleotides derived from nucleotides 199 to 258 of the FR3 gene segments both yields less complex patterns and highlights polymorphic variations between individuals. The potentially important role played by amino acid residues encoded by the FR3 gene segment in influencing conformation, antigen binding and affinity of the physically proximal complementarity determining regions (CDRs) suggests FR3 sequence conservation to be biologically significant. Genomic studies combining information from both CDR and framework restriction fragment polymorphisms can prove useful in detecting specific genes and in identifying haplotypes of interest. PMID- 8418474 TI - Gastric interleukin-8 and IgA IL-8 autoantibodies in Helicobacter pylori infection. AB - Gastric infection with Helicobacter pylori is frequently characterized by neutrophil infiltration. The production of the neutrophil-activating peptide (NAP 1/IL-8) and mucosal IgA autoantibodies to IL-8 by human antral biopsies have been examined during short-term in vitro culture. Detectable IL-8 was secreted by 84% of H. pylori-negative patients with normal antral mucosa (range < 0.07-61.5 ng/mg biopsy protein, n = 19). Concentrations in 4 patients with reactive gastritis and 10 with inactive gastritis were not significantly different from subjects with normal mucosa. In H. pylori-positive patients with active gastritis and neutrophil infiltration into the epithelium (n = 17) IL-8 secretion was significantly increased relative to subjects with normal mucosa (P < 0.0001), inactive gastritis (P < 0.001) and reactive gastritis (P < 0.01). IL-8 concentrations in active gastritis were significantly correlated with the extent of epithelial surface degeneration (r = 0.64). IgA autoantibodies were present in 19 patients (13 active, 4 inactive gastritis) and concentrations were significantly correlated with IL-8 production (P < 0.001). Gastric synthesis of IL-8 is likely to be an important factor in regulating mucosal neutrophil infiltration and activation in patients with H. pylori infection. The local production of IgA antibodies to IL-8 may represent a down-regulatory response of the host to limit mucosal damage associated with a chronic bacterial infection. PMID- 8418473 TI - Membrane-incorporated surrogate receptors increase antigen presentation by resting B cells, but not by LPS-activated blasts. AB - Palmitate-conjugated monoclonal antibodies specific to ovalbumin were inserted into the cell membrane of normal resting B cells and LPS-activated blasts. These two decorated B cells were tested for their ability to act as antigen-presenting cells for ovalbumin-specific I-Ad-restricted T-cell hybridomas. It was found that the antibody-decorated resting B cells presented antigen more efficiently than non-decorated controls. However, no increment was observed when decorated LPS blasts were compared with non-decorated blasts. This is explained by the fact that the inserted antibodies quickly disappeared from the cell membrane of LPS blasts, while they were retained for a long period in the membrane of resting B cells. PMID- 8418472 TI - HLA class II+ human keratinocytes present Mycobacterium leprae antigens to CD4+ Th1-like cells. AB - In a variety of inflammatory skin diseases like leprosy, keratinocytes (KC) are induced to express MHC class II molecules and may therefore serve as antigen presenting cells (APC) for MHC class II restricted T cells infiltrating the lesions. However, KC have been thought to be improper APC for MHC class II restricted T cells and to drive T cells into an anergic rather than into an activation state. We evaluated this issue in relation to leprosy and tested whether HLA-DR+ KC could present M. leprae antigens to well-defined, CD4+, cytotoxic as well as proliferative, Th1-like cell clones. Using a recently developed sensitive assay system which employs intact layers of basal KC as APC we found that most T-cell clones (6/8) lysed HLA-DR+ KC pulsed with M. leprae antigens. KC were only recognized after induction of HLA-DR expression by IFN gamma, in an antigen-specific and HLA class II restricted manner. All T-cell clones tested also showed significant proliferation and IFN-gamma production in response to M. leprae antigens presented by HLA-DR+ KC, arguing against a KC dependent anergizing effect on T cells. Thus, HLA class II+ KC can function as proper APC for HLA class II restricted CD4+ Th 1-like cells. It seems therefore possible that antigen presentation by KC contributes to the local cell-mediated immune responses in DTH lesions. PMID- 8418475 TI - Anti-tumour activity of idiotype-specific, MHC-restricted Th1 and Th2 clones in vitro and in vivo. AB - Idiotypes (Id) can serve as individual markers on B cells; therefore, cytotoxic Id-specific T cells may play a significant role in immunological surveillance of Id+ B-cell tumours. We have investigated the anti-tumour activity of CD4+ BALB/c Th1 and Th2 clones which recognize a processed Id of the syngeneic lambda 2(315) L chain in the context of the class II MHC molecule I-Ed. Id-specific T cells and A20/46 B lymphoma cells transfected with the lambda 2(315) gene were injected s.c. into the same site of BALB/c mice (Winn assay). The results show that both Th1 and Th2 clones can protect against tumour development. The protection was Id specific because T cells did not influence tumour development by an A20/46 B lymphoma cell line transfected with the pSV2neo expression vector alone. In vitro studies showed that the Th1 clones were cytotoxic to lambda 2(315)-transfected B lymphoma cells; by contrast, the Th2 clone was not cytotoxic in 51Cr-release assay even though the Th2 cells inhibited the growth of lambda 2(315) B lymphoma cells. The anti-lymphoma properties of both the Th1 and Th2 clones appear to involve as yet undefined cytotoxic and growth inhibiting molecules. PMID- 8418476 TI - Fibronectin at the lymphocyte surface. Evidence for activation-dependent binding to VLA4 and VLA5 integrins. AB - The surface of in vitro cultured fixed and viable human T lymphocytes and certain T-cell lines was found to react with different monoclonal anti-fibronectin (FN) antibodies as revealed by ELISA, immunocytochemistry and FACS analysis. SDS-PAGE showed that anti-FN antibodies defined a high molecular weight lymphocyte component which could be iodinated using the lactoperoxidase method and which had gelatin binding capacity. FACS analysis showed that the reactivity of anti-FN antibodies with lymphocytes was most pronounced in activated cells and increased during the culture period. By contrast, FACS analysis revealed equal high expression of the VLA4 and VLA5 integrins on freshly purified as well as on mixed lymphocyte culture (MLC) activated cells. Freshly purified lymphocytes and lymphocytes cultured in vitro overnight did not bind 3H-labelled FN in solution whereas MLC-activated cells were capable of 'spontaneous' binding of such [3H] FN. However, brief 12-o-tetradecanoylphorbol-13-acetate (TPA) exposure rendered freshly purified lymphocytes capable of binding soluble FN. These interactions of the lymphocytes with 3H-labelled FN in solution could be almost completely blocked by monoclonal anti-VLA4 and VLA5 antibodies. These results indicate that activated T cells express fibronectin at their surface under 'normal' culture conditions. Although both freshly purified and MLC-activated lymphocytes have equal expression of the integrins VLA4 and VLA5, only activated cells are capable of 'spontaneous' binding FN in solution via an integrin-mediated process, probably via an increase in the affinity of these receptors for FN. PMID- 8418477 TI - Aberrant tolerance induction with cationic antigens. AB - We studied the induction of tolerance in female C57B1/6 mice by oral and intravenous (i.v.) administration of protein antigens before immunization. Native proteins [chicken egg albumin (OVA), bovine serum albumin (BSA)] and their cationic derivatives [amidated (a)OVA, aBSA, methylated (m)BSA] were compared in their capacity to suppress cell-mediated immunity (CMI), as measured by a delayed type hypersensitivity test (DTH). Oral feeding of 0.5 mg negative proteins gave a clear suppression. By contrast, cationic derivatives (50 micrograms to 50 mg) did not suppress CMI. Data from cross-experiments, where aOVA was fed in OVA-immune mice, showed no suppression at all. When OVA was fed in aOVA-immune mice, only a partial suppression was achieved. The potency to induce tolerance differed also when the antigens were administered i.v.: 25 micrograms OVA was sufficient to induce a clear suppression, whereas a much higher amount of aOVA (500 micrograms) caused marginal suppression in OVA- and aOVA-immune mice, respectively. Nevertheless, when concentrations of aOVA up to 2.5 mg were tested in a chronic model of arthritis, a significant suppression was achieved. The differences in inducing a CMI suppression may have implications for the feasibility of immunointervention. Successful modulation of human arthritis may be highly dependent on the nature of the antigen involved. PMID- 8418478 TI - Madagascar's lemurs. AB - These primates can tell us a great deal about our own evolutionary past. But many species are already extinct, and the habitats of those that remain are shrinking fast. PMID- 8418479 TI - Sex on the brain. PMID- 8418480 TI - The mind and Donald O. Hebb. AB - By rooting behavior in ideas, and ideas in the brain, Hebb laid the groundwork for modern neuroscience. His theory prefigured computer models of neural networks. PMID- 8418481 TI - Trends in nonlinear dynamics. Adapting to complexity. PMID- 8418482 TI - Habeas corpus. Seeking subjects to be a digital Adam and Eve. PMID- 8418483 TI - How many genes and Y. Gene mappers find plenty, even in "junk' chromosomes. PMID- 8418484 TI - Endangered genes. PMID- 8418485 TI - A gene for hypertension. PMID- 8418486 TI - Anything goes. Why two sexes are better than 13. PMID- 8418487 TI - Selectivity for polar, hyperbolic, and Cartesian gratings in macaque visual cortex. AB - The neural basis of pattern recognition is a central problem in visual neuroscience. Responses of single cells were recorded in area V4 of macaque monkey to three classes of periodic stimuli that are based on spatial derivative operators: polar (concentric and radial), hyperbolic, and conventional sinusoidal (Cartesian) gratings. Of 118 cells tested, 16 percent responded significantly more to polar or hyperbolic (non-Cartesian) gratings than to Cartesian gratings and only 8 percent showed a significant preference for Cartesian gratings. Among cells selective for non-Cartesian gratings, those that preferred concentric gratings were most common. Cells selective for non-Cartesian gratings may constitute an important intermediate stage in pattern recognition and the representation of surface shape. PMID- 8418489 TI - Health care cost containment: some implications of global budgets. PMID- 8418488 TI - AIDS theories. PMID- 8418490 TI - Human genome program. Healy and Collins strike a deal. PMID- 8418491 TI - Inflamed debate over neurotoxin. PMID- 8418492 TI - Is devastating whitefly invader really a new species? PMID- 8418493 TI - Organic synthesis of prostaglandins: advancing biology. PMID- 8418494 TI - Mathematics achievement of Chinese, Japanese, and American children: ten years later. PMID- 8418495 TI - Selective perhydroxylation of squalene: taming the arithmetic demon. AB - Osmium-catalyzed asymmetric dihydroxylation, which produces 1,2-diols of high enantiopurity from prochiral olefins, is an example of the synthetic catalysts that have been developed that rival enzymes in their efficiency and high enantioselectivity. Although the asymmetric dihydroxylation catalyst lacks an enzyme's ability to effectively distinguish among the subtly different olefinic sites in a polyolefin such as squalene, this very inability permits it to bring about the "exhaustive" polyhydroxylation of squalene to give a dodecahydroxy derivative. Twelve chemical and stereochemical events proceed in tandem with a remarkable average yield of 98 percent per step, giving 1 out of the 36 possible stereoisomers in (0.98)12 = 78.9 percent overall yield. PMID- 8418496 TI - A traveling-wave amplifier model of the cochlea. AB - A two-mode model of the cochlea that uses active intermode feedback has been developed that quantitatively accounts for the motion of the basilar membrane in response to single tones and qualitatively accounts for cochlear emission phenomena. In contrast to existing single-mode models, this model amplifies the mechanical traveling wave in spatially localized cochlear regions where an approximate match occurs between the traveling-wave velocities of each of the two traveling-wave lines or modes. PMID- 8418498 TI - Mediation by G proteins of signals that cause collapse of growth cones. AB - During development, motion of nerve growth cones ceases on contact with particular targets. The signaling mechanism is unknown. In culture, growth cone collapse can be caused by solubilized embryonic brain membranes, central nervous system myelin, a 35-kilodalton protein isolated from myelin, and mastoparan. Collapse induced by each of these is blocked by pertussis toxin. Thus, collapse of growth cones is mediated by G protein-coupled receptors, which may be activated by proteins associated with the cell surface as well as by soluble ligands. PMID- 8418497 TI - Identification of a whitefly species by genomic and behavioral studies. AB - An introduced whitefly species, responsible for over a half billion dollars in damage to U.S. agricultural production in 1991, is morphologically indistinguishable from Bemisia tabaci (Gennadius). However, with the use of polymerase chain reaction-based DNA differentiation tests, allozymic frequency analyses, crossing experiments, and mating behavior studies, the introduced whitefly is found to be a distinct species. Recognition of this new species, the silverleaf whitefly, is critical in the search for management options. PMID- 8418499 TI - Role of intracellular calcium in NI-35-evoked collapse of neuronal growth cones. AB - A myelin-associated protein from the central nervous system, the neurite growth inhibitor NI-35, inhibits regeneration of lesioned neuronal fiber tracts in vivo and growth of neurites in vitro. Growth cones of cultured rat dorsal root ganglion neurons arrested their growth and collapsed when exposed to liposomes containing NI-35. Before morphological changes, the concentration of free intracellular calcium ([Ca2+]i) showed a rapid and large increase in growth cones exposed to liposomes containing NI-35. Neither an increase in [Ca2+]i nor collapse of growth cones was detected in the presence of antibodies to NI-35. Dantrolene, an inhibitor of calcium release from caffeine-sensitive intracellular calcium stores, protected growth cones from collapse evoked by NI-35. Depletion of these caffeine-sensitive intracellular calcium stores prevented the increase in [Ca2+]i evoked by NI-35. The NI-35-evoked cascade of intracellular messengers that mediates collapse of growth cones includes the crucial step of calcium release from intracellular stores. PMID- 8418500 TI - Regulation of the human hsp70 promoter by p53. AB - The tumor suppressor p53 is a nuclear phosphoprotein with characteristics of a transcription factor. It displays sequence-specific DNA binding, contains a potent transactivation domain, and has been implicated as both a transcriptional activator and a repressor. Transcription of the human hsp70 gene is stimulated by adenovirus E1a protein. This E1a transactivation of the hsp70 promoter is mediated by CCAAT binding factor (CBF). It is demonstrated here that p53 both represses transcription from the human hsp70 promoter and also interacts with CBF. Thus, the repression of the hsp70 promoter by p53 may be mediated by direct protein-protein interaction with CBF. These results suggest that protein-protein interaction between p53 and specific transcription factors may be an additional mechanism by which p53 regulates gene expression. PMID- 8418501 TI - Requirement for CD8+ cells in T cell receptor peptide-induced clonal unresponsiveness. AB - T cell receptor (TCR) vaccination in rats prevents the development of experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis. The mechanism of this potential immunotherapy was examined by vaccinating mice with an immunogenic peptide fragment of the variable region of the TCR V beta 8.2 gene. Another immunogen that usually induces an immune response mediated by V beta 8.2+ T cells was subsequently inhibited because specific clonal unresponsiveness (anergy) had been induced. Depletion of CD8+ cells before TCR peptide vaccination blocked such inhibition. Thus, the clonal anergy was dependent on CD8+ T cells, and such immunoregulatory T cells may participate in the normal course of EAE. PMID- 8418502 TI - Treatment and prevention of rat glioblastoma by immunogenic C6 cells expressing antisense insulin-like growth factor I RNA. AB - Rat C6 glioma cells express insulin-like growth factor I (IGF-I) and form rapidly growing tumors in syngeneic animals. When transfected with an episome-based vector encoding antisense IGF-I complementary DNA, these cells lost tumorigenicity. Subcutaneous injection of IGF-I antisense-transfected C6 cells into rats prevented formation of both subcutaneous tumors and brain tumors induced by nontransfected C6 cells. The antisense-transfected cells also caused regression of established brain glioblastomas when injected at a point distal to the tumor. These antitumor effects result from a glioma-specific immune response involving CD8+ lymphocytes. Antisense blocking of IGF-I expression may reverse a phenotype that allows C6 glioma cells to evade the immune system. PMID- 8418503 TI - [Obstruction of the nasal passage caused by a balloon catheter in a calf]. AB - Decompression of the rumen as a supporting symptomatic therapy of recurrent bloat in calves and juvenile cattle is used regularly. The gas should be released for several days. This is facilitated by the use of a balloon stomach tube, which can be left in situ for up to five days. However, the use of this tube is not always without risks, which will be discussed on base of a patient. PMID- 8418504 TI - [Final or interim registered veterinary drugs for processing in medicated feed]. PMID- 8418505 TI - [Diagnosis diarrhea in dogs]. PMID- 8418506 TI - [What is the value of retrieval systems for the veterinarian?]. PMID- 8418507 TI - [Problems and recommendations concerning treatment of exotic pets]. AB - When wanting to treat birds and other unusual companion animals, veterinarians must often refer to data for other animal species and humans. Doses must often be extrapolated from data for other animals, and there is little, or only empirical, information about pharmacology, toxicity, effectiveness, etc. On the basis of what is known, and especially for 'ordinary' companion animals and farm animals, the pharmacokinetics and pharmacodynamics of drugs vary considerably as a result of species differences in drug absorption, distribution, metabolism and elimination. Differences in anatomy, physiology and genetics underlie these variations. For instance, environmental temperature is important in the elimination of drugs by cold-blooded animals. The character and nature of unusual companion animals, their zootechnology, feeding habits, housing requirements etc. often require special and imaginative solutions to the problem of how to administer veterinary medicines. Moreover, when the Veterinary Medicine Act becomes law, terms such as 'off-label use', 'orphan drugs' and 'minor species' will also play a role. PMID- 8418508 TI - [Possible effects on the environment of antibiotic residues in animal manure]. AB - After application to animals veterinary drugs can enter the environment faeces. Little is known about the impact these veterinary drugs can have on the environment. Antimicrobial drugs, ended up in the soil via manure, could affect microorganisms and thus disturb ecological cycles. A study of the literature on the impact on the environment of a number of important (groups of) antibiotics and an antiparasitic agent revealed that little is known about most of these agents. PMID- 8418509 TI - Oil-soluble antioxidants in foods. PMID- 8418510 TI - Overview: mechanisms of antioxidant action on life span. PMID- 8418511 TI - Effects of various antioxidants on aging in Drosophila. PMID- 8418512 TI - Molecular genetic mechanisms in oxidative damage and aging. PMID- 8418513 TI - Genotoxicity of phenolic antioxidants. PMID- 8418514 TI - The importance of cellular proliferation induced by BHA and BHT. PMID- 8418515 TI - Quantitative cancer risk analysis of BHA based on integration of pathological and biological/biochemical information. PMID- 8418516 TI - Modulation of tumor development by butylated hydroxytoluene in experimental animals. PMID- 8418517 TI - Mechanism-based cancer risk assessment of butylated hydroxyanisole. AB - The evidence for BHA-induced carcinogenicity is restricted to the rodent non glandular stomach, which is not found in humans. Although an argument can be made that an effect in this target organ could be indicative of potential carcinogenicity in other tissues, particularly the esophagus, studies have not revealed such a correlation. BHA-induced proliferative effects, which appear to be critical for the tumorigenic response, show a NOEL at a dose below that yielding benign tumors. The NOEL of this BHA-induced proliferative effect demonstrates a substantial margin of safety compared to human intake of BHA. Thus based on both dose-response considerations and the species-specific response, the use of BHA as a food additive does not pose a carcinogenic risk in humans. PMID- 8418518 TI - Antioxidant intake in the U.S. PMID- 8418519 TI - Inhibition of chemical-induced experimental cancer by synthetic phenolic antioxidants. PMID- 8418520 TI - Efficacy of water-soluble analogs of vitamin E in the prevention of myocardial ischemia-reperfusion injury. PMID- 8418521 TI - Antioxidants in lung disease. PMID- 8418522 TI - Oxidation and aging: impact on vision. PMID- 8418523 TI - Antioxidants: the Canadian perspective. PMID- 8418524 TI - Implications for the biomedical field. PMID- 8418525 TI - Biological antioxidant defenses. PMID- 8418526 TI - Antioxidants indigenous to foods. PMID- 8418527 TI - The consequences of free radicals in foods. PMID- 8418528 TI - Paradoxical behavior of antioxidants in food and biological systems. PMID- 8418529 TI - Differentiation between different pathological cerebral embolic materials using transcranial Doppler in an in vitro model. AB - BACKGROUND AND PURPOSE: The detection of circulating particulate cerebral emboli using transcranial Doppler ultrasonography has been recently reported. It has been suggested that this method might allow discrimination between different embolic materials; this would be very useful for selecting specific pharmacological treatment in individual patients. This study was designed to identify those parameters of the Doppler signal that might prove useful in discriminating between different types and sizes of particulate cerebral emboli. METHODS: An extracorporeal circuit filled with a saline/Tween solution and driven by a peristaltic pump was used. The tubing was placed in a skull in the position of the middle cerebral artery. Using transcranial Doppler ultrasound, flow was insonated via the transtemporal window. The following embolic materials of measured sizes (range of maximum dimension, 0.5-5.0 mm) were introduced into the circuit: thrombus (n = 20), platelet-rich aggregates (n = 15), atheromatous material (n = 20), and fat (n = 20). The Doppler signal was recorded during the passage of each embolus. Off-line analysis was performed to measure the maximum amplitude and duration of the signal. RESULTS: For all embolic materials there was a highly significant relation between embolus size and maximum amplitude of the Doppler signal. The closest correlation was obtained when the logarithm of maximum amplitude was used (for thrombi, r = 0.74; for platelet, r = 0.87; for atheroma, r = 0.46; and for fat, r = 0.68). The slope of the regression line differed for the different embolic materials and was significantly steeper for platelets than for atheroma (p < 0.01). Platelet emboli of maximum dimension < or = 1.5 mm resulted in a significantly lower maximum amplitude than similarly sized atheroma emboli (157 dB versus 206.7 dB, p < 0.01). For larger emboli (> 2 mm) there was little further increase in maximum amplitude with increases in embolus size. For all embolic materials there was a highly significant linear relation between embolus size and duration of the high-amplitude (> 150 dB) signal (for thrombi, r = 0.75; for platelet, r = 0.90; for atheroma, r = 0.77; and for fat, r = 0.86). CONCLUSIONS: Platelet emboli result in lower-amplitude signals, and therefore analysis of maximum amplitude may provide information on the type of embolic material. However, it may be difficult to determine whether a given signal is associated with a large platelet embolus or a small atheroma embolus. Duration of the high-amplitude signal will allow accurate estimation of the size of emboli, particularly where the emboli are all of the same material. PMID- 8418530 TI - Effect of oral nimodipine on platelet function. AB - BACKGROUND AND PURPOSE: Nimodipine, a calcium antagonist, has been reported to have beneficial effects in acute ischemic infarction. Some calcium channel antagonists have antiplatelet effects. We investigated the effect of oral nimodipine on platelet function in healthy volunteers. METHODS: Twelve healthy volunteers (6 men and 6 women, mean age 32.9 +/- 5.6 years) took 30 mg nimodipine every 6 hours for 24 hours, followed by a week with no medication, followed by 60 mg every 6 hours for 24 hours. Ex vivo platelet function was measured at baseline, 1 hour after the first dose at each dosage strength, and 1 hour after the last dose at each dosage. Platelet studies included aggregation and adenosine triphosphate release in response to collagen, epinephrine, and adenosine diphosphate; maximal rate of primary aggregation; threshold adenosine diphosphate concentration for second-phase aggregation; and thromboxane B2 release at threshold aggregation. The bleeding time was measured at baseline and after the last 60-mg dose of nimodipine. RESULTS: No change in any platelet function study was seen with 30 mg nimodipine every 6 hours. Platelet function studies were also unchanged after 60 mg every 6 hours, except for a slight decrease in aggregation and adenosine triphosphate release in response to suprathreshold (10 microM) adenosine diphosphate (p = 0.001, Student's paired t test). There was no significant change in bleeding times. CONCLUSIONS: Oral nimodipine has minimal antiplatelet activity in young, healthy subjects. PMID- 8418531 TI - Ipsilateral hypohidrosis in brain stem infarction. AB - BACKGROUND AND PURPOSE: The brain stem is the most important autonomic processing center, but very little attention has been given to clinical manifestations of autonomic failure in brain stem stroke. Our purpose was to evaluate the prevalence, characteristics, and prognostic significance of sweating dysfunction in brain stem infarctions. METHODS: We carried out a prospective study using quantitative evaporimetry to investigate spontaneous and heat-stimulated sweating in 18 healthy control subjects and 18 patients with ischemic brain stem stroke in the acute phase and at 1 and 6 months after infarction. RESULTS: The sweating response induced by a heating stimulus was significantly lower on the ipsilateral side to the infarction than on the contralateral side. Constant ipsilateral hypohidrosis was established in 83% of the patients in the acute phase, in 100% at 1 month, and in 76% at 6 months after infarction. No differences of sweating response were found between medullary and pontine infarcts. CONCLUSIONS: Hypohidrosis throughout the whole ipsilateral side of the body, a long-lasting phenomenon that has not previously been described, is an essential feature of autonomic failure in brain stem infarction. PMID- 8418532 TI - Flow-induced relaxation of the rabbit middle cerebral artery is composed of both endothelium-dependent and -independent components. AB - BACKGROUND AND PURPOSE: The flow-induced relaxation of a branch of the rabbit middle cerebral artery was examined to determine if an endothelial-independent as well as -dependent component occurs in pial as well as systemic small arteries and the possible role of products of the cyclooxygenase and the L-arginine nitric oxide synthase pathways. METHODS: Intraluminal flow was achieved by the infusion of a tissue bath solution into isometrically mounted rabbit pial arteries in a resistance artery myograph through a small pipette. RESULTS: Intraluminal flow caused relaxation of the artery segment precontracted with 10 microM histamine. Treatment of endothelium-intact vessels with the nitric oxide synthase inhibitors NG-nitro-L-arginine (L-NNA) (100 microM) or NG-nitro-L-arginine methyl ester (L NAME) (0.3 mM) significantly reduced the relaxation at flow rates of 5-30 microliters/min. This effect was partially reversed by 1 mM L-arginine. These inhibitors had no effect on the flow-induced relaxation of endothelium-denuded vessels. L-NNA did not influence the relaxation to 1 and 3 microM papaverine. Exposure to 10 microM aspirin, 10 microM indomethacin, or 300 nM tetrodotoxin had no effect on the flow-induced relaxation of either endothelium-intact or -denuded vessels (n = 6). Flow-induced relaxation was attenuated, but not abolished, by removal of the cerebrovascular endothelium. This reduction was not statistically significant. CONCLUSIONS: These results show that intraluminal flow caused relaxation of a branch of the rabbit middle cerebral artery, in part through a mechanism sensitive to inhibitors of nitric oxide synthase, most likely the generation of nitric oxide from the vascular endothelium. The major component of the relaxant response is independent of the endothelium and of nitric oxide synthesis through an L-NNA- or L-NAME-sensitive mechanism. The relaxation does not involve cyclooxygenase products nor neurogenic mediators. These results suggest that pial arteries, like those of the rabbit ear, exhibit a novel mechanism for the flow-induced relaxation of agonist-induced tone that is intrinsic to the tissues of the vascular wall subjacent to the endothelium. PMID- 8418533 TI - Moderate hyperglycemia worsens acute blood-brain barrier injury after forebrain ischemia in rats. AB - BACKGROUND AND PURPOSE: Clinical and experimental data indicate that hyperglycemia can aggravate the consequences of stroke and cerebral ischemia. The purpose of this study was to examine the effects of moderate hyperglycemia on the response of the blood-brain barrier to normothermic (37 degrees C) and hypothermic (30 degrees C) global forebrain ischemia. METHODS: Sixteen rats underwent 20 minutes of four-vessel occlusion followed by 30 minutes of postischemic recirculation. We used the protein tracer horseradish peroxidase as an indicator of increased vascular permeability, and rats were perfusion-fixed for microscopic analysis. To produce moderate hyperglycemia, we gave an intraperitoneal injection of 50% dextrose 15 minutes before the ischemic insult. RESULTS: After normothermic brain ischemia, normoglycemic rats (plasma glucose level, 115 +/- 3 mg/dl) demonstrated extravasated horseradish peroxidase mainly restricted to the cerebral cortex. In contrast, more severe and widespread protein extravasation was documented throughout the neuraxis of hyperglycemic (plasma glucose level, 342 +/- 27) rats. Sites of protein leakage included the cerebral cortex, striatum, hippocampus, thalamus, and cerebellum. Foci of protein extravasation were associated with pial and large penetrating vessels. Intraischemic hypothermia significantly attenuated the blood-brain barrier consequences of hyperglycemic brain ischemia. CONCLUSIONS: Under normothermic ischemic conditions, hyperglycemia significantly worsens the degree of acute blood-brain barrier breakdown compared with normoglycemia. Postischemic blood brain barrier disruption may play an important role in the pathogenesis of increased brain damage associated with systemic hyperglycemia. PMID- 8418534 TI - Effect of brain edema on infarct volume in a focal cerebral ischemia model in rats. AB - BACKGROUND AND PURPOSE: Infarct volume is one of the common indexes for assessing the extent of ischemic brain injury following focal cerebral ischemia. Accuracy in the measurement of infarct volume is compounded by postischemic brain edema that may increase brain volume in the infarcted region. We evaluated the effect of brain edema on infarct volume determined by triphenyltetrazolium chloride and hematoxylin and eosin stains in a focal cerebral ischemia model in rats. METHODS: In a middle cerebral artery occlusion model in rats, infarction is confined to the cerebral cortex. The infarct was delineated by triphenyltetrazolium chloride stain and, in selected samples, by hematoxylin and eosin stain. We determined infarct size at different times after the ischemic insult (6 hours to 7 days) in relation to the evolution of brain edema by the direct measurement of infarct volume. Indirect measurement to reduce the effect of edema on infarct volume was also conducted in the same brain samples. RESULTS: Direct measurement showed that infarct volume fluctuated with the evolution of brain edema (one-way analysis of variance, p < 0.0001). Infarct volume determined by indirect measurement was independent of the extent of brain edema and remained stable from 6 hours to 3 days after ischemia. There was a good correlation between triphenyltetrazolium chloride and hematoxylin and eosin stains in delineating infarct volume with both direct and indirect measurement. CONCLUSION: Traditional direct measurement of infarct volume is associated with an overestimation of infarct volume during the development of brain edema in the first 3 days after ischemia. This artifact can be reduced with indirect measurement, which is based on noninfarcted cortex volume. PMID- 8418535 TI - Autosomal dominant leukoencephalopathy and subcortical ischemic stroke. A clinicopathological study. AB - BACKGROUND AND PURPOSE: We recently described an autosomal dominant syndrome characterized mainly by recurrent strokes and neuroimaging evidence of leukoencephalopathy. We now report the pathological findings in one of the affected subjects. CASE DESCRIPTION: A 40-year-old woman experienced her first grand mal seizure in 1971. From 1983 on she suffered recurrent strokes, seizures, and psychiatric disturbances with depressions, manic episodes, and dementia. In 1988, after her fourth stroke, she became tetraplegic with a severe pseudobulbar palsy, and she died in 1990. Pathological examination disclosed a recent capsulolenticular hematoma, multiple small deep infarcts, a diffuse myelin loss and pallor of the hemispheric white matter, and a widespread vasculopathy of the small arteries penetrating the white matter. The arterial wall was markedly thickened with an extensive nonamyloid eosinophilic deposit in the media and reduplication of the internal elastic lamella. CONCLUSIONS: The underlying lesion of this hereditary disorder is located in the small arteries and is of unknown etiology. It differs from arteriosclerotic and amyloid angiopathies but is similar to that described in some cases of hereditary multi-infarct dementia. PMID- 8418536 TI - Magnetic resonance angiography of cervicocranial dissection. AB - BACKGROUND: A retrospective study of five patients with the clinical or magnetic resonance-based diagnosis of carotid dissection was done. Clinical data, imaging studies, treatment, and outcome were reviewed. The potential applicability of three-dimensional time-of-flight magnetic resonance angiography in these settings was evaluated. SUMMARY OF REPORT: This modality reliably showed vascular abnormalities and focal, segmental, or aneurysmal dilatation when correlated with conventional angiograms in three patients. Turbulence and magnetic susceptibility at the acute turn of the carotid in the petrous canal led to a false-positive diagnosis on magnetic resonance angiography in one patient (in whom subtle fibromuscular hyperplasia was found with conventional angiography but missed with magnetic resonance angiography). CONCLUSIONS: When combined with appropriate clinical signs, magnetic resonance imaging and magnetic resonance angiography can reliably establish the diagnosis of carotid dissection. Pitfalls of magnetic resonance angiography are discussed. PMID- 8418537 TI - Sudden bilateral hearing impairment in vertebrobasilar occlusive disease. AB - BACKGROUND: Bilateral hearing impairment is rare in vertebrobasilar occlusive disease. SUMMARY OF COMMENT: Between 1986 and 1991, we encountered seven patients (four men, three women; median age, 61 years; range, 46-71 years) who had sudden bilateral hearing impairment among 503 patients with vertebrobasilar occlusive disease. The main initial neurological symptoms were sudden bilateral hearing impairment, tinnitus, and vertigo. Acute labyrinthitis or Meniere's disease was the initial diagnosis until subsequent brain stem or cerebellar signs appeared. Brain stem auditory evoked potentials were abnormal bilaterally in six patients but had unilateral attenuation of the IV-V complex in the remaining one patient. Computed tomographic scans in all six patients showed multiple hypodense lesions in the brain stem and the cerebellum. Cerebral angiography showed complete occlusion on both vertebral arteries in one patient, occlusion on the left with small caliber on the right in another, and severe stenosis on both sides in a third. There was no opacification of internal auditory arteries in these three patients. The remaining patient had arteriosclerotic changes with faint opacification of the bilateral internal auditory arteries. Five patients had a poor prognosis, with locked-in state in four and severe truncal ataxia in one. CONCLUSIONS: We conclude that sudden bilateral hearing impairment in vertebrobasilar occlusive disease is more common than previously recognized and that it may indicate a grave prognosis. PMID- 8418538 TI - Biologic responses to antithrombotic, anticoagulant, or fibrinolytic agents may differ in elderly patients. PMID- 8418539 TI - In vivo antithrombotic effect of aspirin: dose versus nongastrointestinal bleeding. PMID- 8418540 TI - Stroke risk factors in an African population: a report from Sierra Leone. PMID- 8418541 TI - Platelet-activating factor acetylhydrolase in red cell membranes. Does decreased activity impair erythrocyte deformability in ischemic stroke patients? AB - BACKGROUND AND PURPOSE: Platelet-activating factor acetylhydrolase hydrolyzes platelet-activating factor (1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine). It also hydrolyzes oxidized derivatives of phosphatidylcholine that have a short chain acyl residue at the sn-2 position. This enzyme may act mainly in the degradation of oxidized phospholipids and may play a role in maintaining erythrocyte deformability. Therefore, we assessed the activity of red cell membrane platelet-activating factor acetylhydrolase in patients with ischemic stroke and studied the relation of the enzyme activity to red cell deformability. METHODS: Enzyme activity was measured in the detergent extract of red cell membranes from 38 patients with cerebral thrombosis and 38 age-matched healthy volunteers. Red cell filterability, an index of red cell deformability, was also measured. RESULTS: The enzyme activity in patients and control subjects was 100 +/- 74 and 148 +/- 128 nmol/g protein per minute (2.68 +/- 2.11 and 3.79 +/- 2.46 pmol/10(9) cells per minute) (mean +/- SD), respectively, and the difference was significant (p < 0.05 by the Mann-Whitney U test, two-sided test). Enzyme activity was correlated positively with red cell filterability in the patients (n = 20, r = 0.565, p < 0.01). CONCLUSIONS: Red blood cells from stroke patients have lower levels of platelet-activating factor acetylhydrolase activity when compared with those from healthy subjects. This may result in the accumulation of oxidized lipids in the cell membrane and lead to impaired red cell deformability in patients with cerebral thrombosis. PMID- 8418542 TI - Wallenberg's syndrome secondary to bullet injury of the vertebral artery. PMID- 8418543 TI - Pure motor monoparesis due to intracerebral hemorrhage. PMID- 8418544 TI - Hypoglycemia presenting as acute tetraplegia. PMID- 8418545 TI - Ischemic stroke due to deficiency of coagulation inhibitors. Report of 10 young adults. AB - BACKGROUND AND PURPOSE: Deficiencies in coagulant inhibitors protein C, protein S, and antithrombin III increase the risk of venous thrombosis. We describe 10 young adults with cerebral arterial thrombosis due to deficiencies in these factors. METHODS: Sixty patients younger than 45 years were hospitalized because of acute ischemic stroke diagnosed through computed tomography or magnetic resonance imaging. Cerebral angiography was performed in 54 cases. Hematologic and coagulation profiles, autoantibody screen, syphilis serology, and lupus anticoagulant were analyzed in all patients. Among the total cases, Holter monitoring was performed in 13 patients, echocardiography in 20, and cerebrospinal fluid studies for cysticercosis and tuberculosis in two. The quantitative analysis of protein C, protein S (by Laurell rocket immunoelectrophoresis), and antithrombin III (by radial immunodiffusion) was performed on admission and 3 months after stroke in all patients and in relatives of six patients. RESULTS: In 10 cases (17%) the stroke was attributed to deficiency of coagulation inhibitors: three men had protein C deficiency, two women had protein S deficiency, and five had antithrombin III deficiency (three men and two women). The cerebral infarction involved the carotid territory in these 10 patients. None had previous thromboembolic disease. Eight patients showed a complete recovery. An acquired disorder was presumed in one protein S deficient and in two antithrombin III-deficient patients; the remainder were considered heterozygous. CONCLUSIONS: The cerebral vasculature may be primarily involved in the deficiency of these natural anticoagulants. Young adults seem to be the most often affected. A knowledge of these new clotting defects will enable the clinician to improve the prevention and treatment of this devastating neurological disease. PMID- 8418546 TI - Atrial fibrillation after acute stroke. AB - BACKGROUND AND PURPOSE: Atrial fibrillation (AF) is a risk factor for stroke, although it may not always be directly responsible for the stroke. On the other hand, cardiac arrhythmias and electrocardiographic changes have been reported after ischemic stroke and numerous other intracranial pathologies. We tested the hypothesis that some patients with acute stroke may develop transient AF as a consequence of the stroke. METHODS: This study was based on 1,661 patients with first-ever stroke consecutively hospitalized and prospectively included into the Lausanne Stroke Registry. "Recent AF" was defined as AF discovered at or after ("after-admission" AF) admission in patients without any previous history of AF. Populations with recent AF and after-admission AF were compared for AF evolution, risk factors, and lesion type and distribution with patients with previous history of AF (known AF) and with patients with another recognized cardiac source of embolism (cardioembolic). RESULTS: Twenty-four patients had recent AF on admission, and 17 developed it a few hours to 3 days after stroke. AF disappeared after a few days in 26 (63%; 94% of after-admission AF) patients. Stroke was a primary hematoma in 9.8% of patients with recent AF, 2.8% of patients with known AF, and 0.9% (p < 0.001) of patients with cardiac source of embolism. Parietoinsular (32%) and brain stem (11%) involvement were more common in recent AF than in cardioembolic stroke in general (16.7% and 6.7%, respectively; p < 0.05). CONCLUSIONS: AF discovered after an acute stroke lasted no more than a few days, suggesting that it may have occurred as a consequence of the stroke. This possibility is emphasized by the significant predominance in patients with recent AF of primary hematoma, which cannot be caused by AF, and of parietoinsular and brain stem involvement, which are experimentally known as arrhythmogenic. This hypothesis should be considered in patients with acute stroke and previously unknown AF before therapeutic decisions are made. PMID- 8418547 TI - Patent foramen ovale: is stroke due to paradoxical embolism? AB - BACKGROUND AND PURPOSE: A patent foramen ovale has been reported to be significantly more frequent in young stroke patients than in matched control subjects, and paradoxical embolism has been suggested as the main mechanism of stroke in this situation. The present study was designed to test this hypothesis. METHODS: Sixty-eight consecutive patients under 55 years of age presenting with an ischemic stroke had an extensive workup, including transesophageal echocardiography with contrast. We compared the prevalence of criteria for the diagnosis of paradoxical embolism in patients with and without a patent foramen ovale. RESULTS: A patent foramen ovale was found in 32 patients (47%). A Valsalva provoking activity was present at stroke onset in six patients with a patent foramen ovale and in eight patients with no patent foramen ovale (chi 2 = 0.1, nonsignificant). Clinical/radiological features suggestive of an embolic mechanism were not more frequent in patients with a patent foramen ovale. Clinical evidence of deep vein thrombosis was present in one patient with a patent foramen ovale and in none of the others. No occult venous thrombosis was found in a subgroup of patients with a patent foramen ovale and no definite cause for stroke who underwent venography (n = 13). CONCLUSIONS: Our results do not support the hypothesis that paradoxical embolism is the primary mechanism of stroke in patients with a patent foramen ovale. PMID- 8418548 TI - Multivariate prediction of the first major cerebrovascular event in an Italian population sample of middle-aged men followed up for 25 years. AB - BACKGROUND AND PURPOSE: The present investigation was aimed at evaluating the incidence and prediction of a first major cerebrovascular (fatal or nonfatal) event. METHODS: The study population included the two Italian rural samples of the Seven Countries Study (namely, Montegiorgio and Crevalcore), accounting for a total of 1,712 men aged 40-59 years at entry and followed up for mortality and morbidity for 25 years. A number of individual variables measured at baseline, at the fifth year, and at the tenth year of follow-up and possibly related to cerebrovascular events were considered. Of the 1,709 subjects free from major cerebrovascular events at entry 171 developed a first major cerebrovascular event, but for the multivariate Cox model analysis only 1,572 subjects and 152 events were employed due to some exclusions for missing data. RESULTS: Systolic blood pressure, indexes of respiratory function (protective), and physical activity at work (protective) demonstrated significant predictive roles for all ages and all lengths of follow-up considered. Other factors (presence of arrhythmias, presence of arcus senilis, and skinfold thickness [protective]), significantly contributed to the prediction, but in only some models. Time related changes in systolic blood pressure significantly improved the prediction of cerebrovascular events. CONCLUSIONS: The multivariate prediction performed in this report allowed the validation of three risk factors (systolic blood pressure, respiratory function indexes, and physical activity at work) whose predictive powers remain stable with aging. The need for further studies specifically aimed at discriminating hemorrhagic from thrombotic events is suggested. PMID- 8418549 TI - Lobar hemorrhage in the elderly. The undiminishing importance of hypertension. AB - BACKGROUND AND PURPOSE: We sought to determine whether hypertension is less common in primary lobar hemorrhage than intracerebral hemorrhage in other locations and whether the frequency of hypertension in lobar hemorrhage diminishes with advancing age. METHODS: We identified all cases of intracerebral hemorrhage in Greater Cincinnati during 1988 by review of hospital and autopsy records as well as computed tomographic and magnetic resonance scans. RESULTS: During 1988, 66 primary lobar hemorrhages occurred, constituting 46% of all intracerebral hemorrhages in those under 75 years of age and 34% in those age 75 and older. A history of hypertension was present in 67% of patients with lobar, 73% of those with deep, 73% of those with cerebellar, and 78% of those with pontine hemorrhages. Left ventricular hypertrophy was present in 21% of patients with lobar, 27% of those with deep, and 47% of those with pontine/cerebellar hemorrhages. The frequency of hypertension in patients with lobar hemorrhage did not decrease with advancing age. CONCLUSIONS: The proportion of all intracerebral hemorrhages that are lobar does not increase with advancing age. Hypertension is nearly as common in primary lobar hemorrhage as in deep, cerebellar, and pontine hemorrhages, and its importance as an associated condition for lobar hemorrhage does not diminish with advancing age. PMID- 8418550 TI - Different linkage of depression to hypercortisolism early versus late after stroke. A 3-year longitudinal study. AB - BACKGROUND AND PURPOSE: Using the dexamethasone suppression test, we studied the suppressibility of the cortisol axis and its clinical determinants at various time points after stroke. A major aim was to examine the dexamethasone test as a diagnostic tool for the diagnosis of major depression in stroke patients. METHODS: The dexamethasone suppression test, major depression, functional ability, and disorientation were assessed in a cohort of 70 patients with acute stroke and after 3 months (n = 63) and 3 years (n = 43). RESULTS: Early after stroke, 24% of the patients were nonsuppressors, with about the same proportion at 3 months (22%) and 3 years (21%). None of the controls (17 healthy elderly volunteers) were nonsuppressors. High cortisol levels early after stroke were significantly associated with functional impairment (r = 0.35; p = 0.003) and disorientation (r = 0.27; p = 0.03). Three years after stroke, high postdexamethasone cortisol levels were significantly associated with major depression (r = 0.57; p < 0.001). The sensitivity of the dexamethasone test was 70% and the specificity 97%. In a longitudinal analysis of the long-term survivors (n = 42), postdexamethasone cortisol values at 3 months predicted major depression at 3 years. CONCLUSIONS: Hypercortisolism is associated with major depression late (3 years) but not early (0-3 months) after stroke. Patients with hypercortisolism 3 months after stroke are at risk of major depression later in the course and warrant careful follow-up from a psychiatric viewpoint. PMID- 8418551 TI - Measuring physical impairment and disability with the Chedoke-McMaster Stroke Assessment. AB - BACKGROUND AND PURPOSE: The Chedoke-McMaster Stroke Assessment measures the physical impairments and disabilities that impact on the lives of individuals with stroke. This measure has three overall purposes: 1) to stage motor recovery to classify individuals in terms of clinical characteristics, 2) to predict rehabilitation outcomes, and 3) to measure clinically important change in physical function. This study was carried out to evaluate the ability of this measure to yield reliable and valid results. METHODS: Thirty-two subjects from a stroke rehabilitation treatment unit were assessed by research and treating physical therapists using multiple measures on multiple occasions. The measure's three purposes dictated the study objectives and design. RESULTS: Intrarater, interrater, and test-retest reliabilities of the impairment and disability inventories were estimated. Reliability coefficients for the total scores ranged from 0.97 to 0.99. Construct and concurrent validities were studied by examining the correlations between this and other measures. A priori hypothetical constructs stated that these correlations should exceed 0.60. These constructs were confirmed; the impairment inventory total score was found to correlate with the Fugl-Meyer Test (r = 0.95, p < 0.001) and the disability inventory with the Functional Independence Measure (r = 0.79, p < 0.05). Additional study hypotheses were also substantiated. CONCLUSIONS: This study confirms that the Chedoke McMaster Stroke Assessment yields both reliable and valid results. With the evaluation study now completed, the Chedoke-McMaster Stroke Assessment can be used with confidence as both a clinical and a research tool that can discriminate among subjects and evaluate patient outcomes. PMID- 8418552 TI - Amplitude of the ocular pneumoplethysmography waveform is correlated with cardiac output. AB - BACKGROUND AND PURPOSE: Ocular pulse amplitude, the amplitude of the ocular pneumoplethysmographic waveform, is altered in several ophthalmologic diseases that disturb ocular blood flow, implying that ocular pulse amplitude may provide an estimate of ocular blood flow. Because ocular blood flow currently cannot be quantified in humans, two experiments were undertaken to evaluate the association of ocular pulse amplitude with total body blood flow. METHODS: In experiment 1, cardiac output was determined by cardiac catheterization in 181 patients who underwent OPG-Gee testing during the same hospitalization. In experiment 2, 110 instances of atrial arrhythmia captured on ocular pneumoplethysmographic tracings were evaluated for transient changes in heart rate (R-R ratio) associated with transient changes in ocular pulse amplitude (ocular pulse amplitude ratio). RESULTS: In experiment 1, average ocular pulse amplitude in the two eyes (OPAAV) was significantly correlated with cardiac output/heart rate (r = 0.53; p < 0.0001) and cardiac index/heart rate (r = 0.43; p < 0.0001). In experiment 2, R-R ratio was significantly correlated with ocular pulse amplitude ratio (r = 0.85; p < 0.001). CONCLUSIONS: These results show that ocular pulse amplitude, a physiological measurement obtained from the globe, is correlated with cardiac output. They imply that ocular pulse amplitude may provide a clinically useful estimate of at least the pulsatile component of ocular blood flow. PMID- 8418553 TI - Impact of social support on outcome in first stroke. AB - BACKGROUND AND PURPOSE: The purpose of this study is to examine the impact of social support on outcome after first stroke in a prospective cohort study. Although modest evidence exists for the importance of several psychosocial factors, studies have failed to use widely recognized measures of outcome and social support, have failed to control for time since onset, and have not used longitudinal techniques. METHODS: Forty-six surviving patients were followed for 6 months after stroke. Recovery was measured using repeated measures of functional status as indicated by the Barthel Index of activities of daily living. Perceived social support was measured at 1, 3, and 6 months after onset. Repeated-measures multivariate analysis of variance was used to analyze changes in functional status. RESULTS: Significant differences were found across levels of social support in trajectories of functional status (p = 0.002). A significant three-way interaction between stroke severity, social support, and outcome was also found (p = 0.012). Patients with more severe stroke and the largest amount of social support attained an average Barthel Index that was 68 points (65%) higher than the group reporting the least support. CONCLUSIONS: High levels of social support were associated with faster and more extensive recovery of functional status after stroke. Social support may be an important prognostic factor in recovery from stroke. Socially isolated patients may be at particular risk for poor outcome. PMID- 8418554 TI - Decline in autopsies for deaths attributed to cerebrovascular disease. AB - BACKGROUND AND PURPOSE: United States national autopsy rates have declined in recent years. In the present study, changes in autopsy rates for deaths due to stroke are examined and compared with changes in autopsy rates for all deaths. METHODS: National Center for Health Statistics data on United States national autopsy rates were examined for the years 1955, 1958, and 1972-1988. RESULTS: Since at least 1955, nonstroke deaths were more than twice as likely to be autopsied as deaths due to cerebrovascular disease. The annual autopsy frequency for all deaths, for deaths due to stroke, and for deaths due to each stroke subtype declined precipitously after 1972. Since 1982, less than 5 percent of deaths attributed to stroke have been documented by autopsy. Information obtained at autopsy was frequently ignored in the determination of cause of death on the death certificate. CONCLUSIONS: Careful consideration of the value of autopsy for education, research, and quality assurance is urgently needed. Unless the present problems in obtaining, processing, disseminating, and using autopsy data are adequately addressed, the autopsy rate will continue to decline. PMID- 8418555 TI - Cerebellar infarction. Clinical and anatomic observations in 66 cases. AB - BACKGROUND AND PURPOSE: Cerebellar infarction displays different clinical features, depending on the vascular territory involved. We studied patients with infarcts in the territories of the posterior inferior cerebellar artery or the superior cerebellar artery to compare their clinical presentation, course, and prognosis. METHODS: We retrospectively analyzed the clinical features, laboratory data, and imaging studies of 66 patients with cerebellar infarction collected consecutively at five institutions. All the cerebellar infarcts were documented on computed tomographic scan or magnetic resonance imaging. RESULTS: Two distinct profiles emerged, depending on the vascular territory involved. In 36 patients with posterior inferior cerebellar artery territory infarcts, a triad of vertigo, headache, and gait imbalance predominated at stroke onset. Computed tomography showed severe cerebellar mass effect in 11 cases (30%), with associated hydrocephalus in seven. In these seven patients (19%), postinfarct swelling led to brain stem compression that resulted in four deaths. In 30 patients with superior cerebellar artery infarcts, gait disturbance predominated at onset; vertigo and headache were significantly less common. The clinical course was usually benign. Computed tomography showed marked cerebellar mass effect, hydrocephalus, and brain stem compression in only two instances (7%). Presumed cerebral embolism was the predominant stroke mechanism in patients with superior cerebellar artery distribution infarcts, whereas in those with posterior inferior cerebellar artery distribution infarcts, the stroke mechanism was equally divided between cardiogenic embolism and posterior circulation arterial disease. CONCLUSIONS: Cerebellar infarcts in the posterior inferior cerebellar artery and superior cerebellar artery distribution have distinct differences in clinical presentation, course, and prognosis. These differences should help in the selection of appropriate monitoring and treatment strategies. PMID- 8418556 TI - Hypoperfusion and vasoreactivity in the thalamus and cerebellum after stroke. AB - BACKGROUND AND PURPOSE: Cerebellar hypoperfusion in the contralateral hemisphere after stroke is well studied and termed crossed cerebellar diaschisis. However, studies of hypoperfusion in the ipsilateral thalamus have been few. The purpose of this study was to investigate the prevalence of hypoperfusion and vasoreactivity to acetazolamide in the thalamus and cerebellum after stroke. METHODS: We studied cerebral blood perfusion in the thalamus and cerebellum of 14 patients with unilateral cerebral infarction using [123I]isopropyliodoamphetamine single-photon emission computed tomography (123I-IMP SPECT). We also administered acetazolamide-stressed 123I-IMP SPECT to determine vasodilator capacity in these two areas. Regions of interest were drawn over the bilateral thalami and bilateral cerebellar cortices, and asymmetry indexes were obtained. RESULTS: We found ipsilateral thalamic hypoperfusion in 12 (85.7%) and contralateral cerebellar hypoperfusion in 11 (78.6%) of 14 patients. Hypoperfusion was improved (p < 0.01 by the Wilcoxon signed rank test) in 11 (91.7%) of the 12 patients with ipsilateral thalamic hypoperfusion and in seven (63.6%) of the 11 patients with contralateral cerebellar hypoperfusion. CONCLUSIONS: Hypoperfusion in the ipsilateral thalamus and contralateral cerebellum is common, and vasoreactivity to acetazolamide is preserved in both the thalamus and the cerebellum with hypoperfusion. PMID- 8418557 TI - Callosal atrophy with reduced cortical oxygen metabolism in carotid artery disease. AB - BACKGROUND AND PURPOSE: In cerebrovascular disease, brain atrophy may be a reflection of ischemic changes. Minor changes that are not detectable as infarction on computed tomography may result in atrophy with metabolic depression. We evaluated the relation between size of the corpus callosum and cerebral cortical oxygen metabolism in carotid artery disease. METHODS: We used magnetic resonance imaging and positron emission tomography to evaluate 13 right handed male patients with transient ischemic attacks or minor strokes and unilateral internal carotid artery occlusive disease, two with stenosis and 11 with occlusion. Computed tomography showed only minor subcortical abnormalities in the affected hemisphere. Midsagittal corpus callosum areas on T1-weighted magnetic resonance images, white matter lesions on T2-weighted images, and the cerebral metabolic rate of oxygen measured by the oxygen-15 steady-state technique were analyzed. RESULTS: Compared with 13 age- and sex-matched control subjects showing the same degree of white matter areas of high intensity, the patients had significantly decreased callosal areas. The degree of corpus callosum atrophy was significantly correlated with the mean cortical oxygen metabolic rate. The same tendency was also found in five patients without infarcts on computed tomography. However, neither the callosal area nor the hemispheric metabolic rate was correlated with the extent of white matter lesions. CONCLUSIONS: These findings indicate that atrophy of the corpus callosum occurs in association with a decrease in cortical oxygen metabolism. Corpus callosum atrophy may be a useful morphological index that reflects the cerebral cortical metabolic state in carotid artery occlusive disease. PMID- 8418558 TI - Regional cerebral blood flow in chronic stroke patients. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate regional cerebral blood flow parameters during the postacute phase of unilateral ischemic stroke and to correlate them with clinical data. METHODS: Regional cerebral blood flow was measured in 187 patients in the stabilized phase of stroke by the xenon 133 inhalation method with 32 extracranial detectors. Thirty-eight patients were reexamined after a mean +/- SD time of 32 +/- 21.4 months. RESULTS: The overall detection of hypoperfusion was 92.0%, with asymmetries as the most sensitive index, especially for patients with a lesser degree of neurological disability. Neurological disability score was strongly associated with regional cerebral blood flow in the affected hemisphere (p < 0.0001) and with asymmetries (p < 0.0001). The presence of carotid obstruction further decreased the regional cerebral blood flow in the affected hemisphere and significantly increased asymmetry (p < 0.0001). Subjects who had no hypoperfusion at absolute values analysis were more frequently free of carotid disease and had less severe disability than those who had bilateral or unilateral regional absolute cerebral blood flow reduction. In 38 patients without new cerebrovascular events, a significant (p = 0.005) reduction of hemispheric regional cerebral blood flow asymmetries was found on a follow-up examination. CONCLUSIONS: These data confirm the value of regional cerebral blood flow asymmetries in stroke detection and point out that important clinical information is also contained in absolute values analysis. PMID- 8418559 TI - Acoustic non-linearity method for estimating the ratio of bound to free water of biological media. AB - The acoustic non-linear properties of bound and free water in biological media are treated based on their structural differences. A model describing the different effects of bound and free water in protein-water solutions and in biological media is proposed. The ratio of bound to free water can be estimated from this model from measurements of the acoustic non-linearity parameter B/A, the sound speed, and the density of the specimens. PMID- 8418560 TI - Extra-anatomic bypass: a new look (supporting view). PMID- 8418561 TI - Extra-anatomic bypass: a new look (opposing view). AB - The criteria by which revascularizations are measured ultimately is patency, since cessation of blood flow equates with a failed graft and usually signals a return to the preoperative limb perfusion or less. An uncommonly analyzed criterion for graft function is blood flow capacity through the reconstruction which is critically important when increased blood flow demands are required with exercise. This is particularly true in the aortoiliac segment when the bypass must meet large blood flow requirements. When the extra-anatomic bypass is evaluated in this manner, it is often inadequate to meet the blood flow demands required with activity. The source of blood flow is the subclavian artery and not the aorta. The graft diameter is almost one half that used to bypass or substitute for the aorta and four times as long. It is not surprising, therefore, that increases in blood flow are limited in this remote subcutaneous bypass circuit. The axillofemoral and, to some extent, the femorfemoral bypass grafts produce hemodynamic gradients to blood flow increases above basal rates. Thus, graft patency alone does not realistically measure the functional capacity of an aortoiliac reconstruction. The conceptual design of an extra-anatomic bypass results in hemodynamic liabilities that produce an inferior performance to that observed with in-line direct aortofemoral bypass. This observation further supports our contention that extra-anatomic bypass of the aortoiliac segment should be restricted to circumvent a prosthetic graft infection in the aortoiliac segment to maintain limb perfusion following graft removal. Whether it has a role in selected patients with a short life span and critical ischemia who appear to be a prohibited risk for in-line aortofemoral reconstruction will always be debatable. Current experience does not justify expanded indications for extra anatomic bypass in symptomatic patients with aortoiliac disease. The proven effective and durable intervention is aortofemoral revascularization. PMID- 8418562 TI - An update on pancreas transplantation. PMID- 8418563 TI - Segmental liver resection. PMID- 8418564 TI - Living related donors for liver transplants. PMID- 8418565 TI - Portal hypertension and shunt surgery. AB - Surgical shunts in the management of portal hypertension are one component of overall management. The main complications of portal hypertension is variceal bleeding. The very high mortality rate of this complication mandates careful assessment and appropriate treatment. Despite the introduction of new treatment methods such as pharmacologic reduction of portal pressure and endoscopic sclerotherapy, taken in conjunction with the coming of age of liver transplantation, surgical decompressive shunts still have a role in the overall treatment of these patients. The choice of therapy is based on full evaluation. The emphasis of that evaluation is on hepatic function. The patients who can live with their liver for an acceptable period of time and do not have the bleeding controlled with lesser medical managements require decompressive shunts. Surgical decompression of portal hypertension may also be required in the patient who is bleeding to death in the acute setting and, finally, is required for the treatment of acute Budd-Chiari syndrome. The choice of shunt procedures should take into account the potential for liver transplantation at some point in the future. PMID- 8418566 TI - Open human fetal surgery. PMID- 8418567 TI - Surgery for duodenal and gastric ulcer disease. PMID- 8418568 TI - Surgical management of Crohn's disease. PMID- 8418569 TI - Multiple organ failure. PMID- 8418570 TI - The pathogenesis of gallstone formation. PMID- 8418571 TI - Laparoscopic appendectomy and herniorrhaphy. PMID- 8418572 TI - Endoscopic esophagectomy. PMID- 8418573 TI - Advances in management of abdominal aortic aneurysm. PMID- 8418574 TI - Carotid endarterectomy. PMID- 8418575 TI - Diary of a week in practice. PMID- 8418576 TI - Vaginal birth after cesarean section: the patient's point of view. AB - Advocacy of vaginal birth after cesarean section (VBAC) is the current standard of care. We interviewed patients in our program about their attitudes toward VBAC and cesarean section. The success rate in our VBAC program is similar to rates reported in the literature (65 percent). However, interviews with our patients revealed that 40 percent had no desire to participate in the VBAC program, although they fulfilled the criteria for eligibility. The main reasons given for declining a trial of VBAC were the convenience of an elective cesarean section and fear of another prolonged, painful and potentially dangerous labor. Thirty two percent of patients in whom VBAC was successful were dissatisfied with the experience and would have preferred an elective cesarean section. The reasons patients gave for attempting VBAC were different from the medical reasons proposed to them. The main reasons given were a desire to deliver "naturally," a fear of surgery and the concern that cesarean section might harm them or their baby. PMID- 8418577 TI - Promoting vaginal birth after cesarean section. AB - New data support the safety of vaginal birth after cesarean section (VBAC). Physicians and hospitals that provide standard obstetric care can also provide care for women attempting VBAC. The management of labor in such patients differs little from that in other patients, and an understanding of the normal course of labor in these patients should help eliminate inappropriate interventions. Family physicians can play a major role in promoting VBAC in appropriately selected patients. PMID- 8418578 TI - Head-upright tilt test: a new method of evaluating syncope. AB - Syncope and near-syncope account for up to 600,000 visits to emergency departments or physicians' offices each year. Syncope is a prognostic indicator of early mortality. Although $750 million is spent each year to evaluate patients with syncope, 40 to 50 percent of cases elude diagnosis. The head-upright tilt test is a new tool for evaluating patients who are susceptible to vasovagal syncope. In susceptible patients, this test may reproduce the symptoms and signs of previous syncopal episodes. It can also help differentiate convulsive syncope from epilepsy. Patient education, medical therapy and even pacemaker placement may be effective in controlling the episodes in patients with recurrent vasovagal syncope. PMID- 8418579 TI - Methanol toxicity. Agency for Toxic Substances and Disease Registry. AB - Methanol is used in a variety of commercial and consumer products. Increased use of methanol as a motor fuel may lead to higher ambient air levels and a greater potential for ingestion from siphoning accidents. Methanol toxicity initially is not characterized by severe toxic manifestations. Pathophysiologically, methanol toxicity represents a classic example of "lethal synthesis," in which toxic metabolites can cause fatality after a characteristic latent period. Methanol is well absorbed following inhalation, ingestion or cutaneous exposure. It is oxidized in the liver to formaldehyde, then to formic acid, which contributes to the profound metabolic acidosis occurring in acute methanol poisoning. The metabolic products of methanol can produce a syndrome of delayed-onset acidosis, obtundation, visual disturbance and death. Intravenous sodium bicarbonate therapy should be considered if the patient's blood pH is below 7.2. Symptoms and history determine whether intravenous ethanol therapy and hemodialysis should be instituted. PMID- 8418580 TI - Osteopetrosis ('marble bone' disease). AB - Osteopetrosis is a hereditary disorder in which pathologic alteration of osteoclast resorption of bone results in thickening of cortical and lamellar bone. Before bone marrow transplantation, the infantile recessive form was uniformly fatal within the first two decades of life as a result of invasion of the marrow space by abnormal bone formation. The adult autosomal dominant form causes minimal morbidity and is usually diagnosed incidentally on routine radiographs. Although osteopetrosis is an extremely rare disorder, the study of this disease can provide insights into the formation of bone and the inheritance of disease. PMID- 8418581 TI - Anterior knee pain: the challenge of patellofemoral syndrome. AB - Pain in the anterior region of the knee is one of the most common musculoskeletal complaints in children, adolescents and adults. Difficult and frustrating for the patient to endure, the disorder is also difficult for the physician to manage. The precipitating events and the location and duration of the pain are important factors in narrowing the differential diagnosis. A detailed physical examination further aids in the differential diagnosis. The treatment and rehabilitation of patellofemoral dysfunction, the most common cause of anterior knee pain, has four phases--acute, subacute, chronic and maintenance. These phases involve medication, a structured knee exercise program and activity modification before returning to normal activity. More than 80 percent of patients with patellofemoral dysfunction respond well to a nonsurgical approach. PMID- 8418582 TI - Antipsychotic agents: a review. AB - Antipsychotic agents are useful in the treatment of psychosis due to both functional disorders (i.e., idiopathic disorders that are usually treated by psychiatrists) and organic mental disorders. These drugs are classified as low- and high-potency agents. Low-potency agents such as chlorpromazine block muscarinic and alpha 1-adrenergic receptors. Consequently, they produce anticholinergic side effects and orthostatic hypotension. High-potency antipsychotic agents have a higher affinity for dopamine receptors and a relatively negligible affinity for muscarinic and alpha 1 receptors. The high potency agents frequently cause extrapyramidal side effects, such as dystonia and parkinsonism. Serious reactions to antipsychotic drugs include tardive dyskinesia and neuroleptic malignant syndrome. PMID- 8418583 TI - AAP issues guidelines for preventing group B streptococcal infections. PMID- 8418584 TI - The case for family practice and primary care research at the NIH. PMID- 8418585 TI - Corporal punishment. PMID- 8418586 TI - Gynecologic malignancy presenting as hernia. PMID- 8418587 TI - Disulfiram for alcoholism. PMID- 8418588 TI - Treatment of verruca acuminata. PMID- 8418589 TI - Traumatic death in urban children, revisited. AB - OBJECTIVE: Trauma is the leading cause of death in children older than 1 year in the United States. We performed an analysis of the causes of death due to trauma in children in a large urban community to suggest means of prevention in such communities. We also examined data obtained before and after the designation of pediatric trauma centers to determine whether this has made a difference. DATA SOURCES: Records of the Medical Examiner, Cook County, Illinois, from 1983 through 1988. STUDY SELECTION: The admitting log was reviewed for all children before their 16th birthday. During the 6-year study period, 3121 autopsies were performed on children, 36.1% of whom died due to traumatic injuries. We reviewed the records of those children who died secondary to these injuries. DATA EXTRACTION: Record review on pediatric trauma deaths as to cause of death, time of death, age, sex, and any other pertinent information. RESULTS: Of all trauma deaths, fire was the most common cause of death, followed by motor vehicle related injuries, homicides, drownings, and falls. These findings differ from national statistics. Improvement in outcome was seen following the designation of general trauma centers, with further improvement seen following the designation of specific pediatric trauma centers. CONCLUSIONS: Identification of causes of pediatric trauma death enables us to suggest methods of prevention. The centralized care of seriously injured children through the establishment of trauma centers and, specifically, pediatric trauma centers might help to prevent these deaths. Further study of pediatric trauma deaths, including hospital and ambulance records, is needed to improve medical care. PMID- 8418590 TI - Evaluating the effects of early intervention. A mismatch between process and product? PMID- 8418591 TI - Risk factors in febrile seizures. Any surprises? PMID- 8418592 TI - Accuracy of documented vaccination status of patients in pediatric emergency departments. PMID- 8418593 TI - Invasive Pseudomonas infection in two healthy children following prolonged bathing. PMID- 8418594 TI - Cardiac anomalies in ataxia-telangiectasia. PMID- 8418596 TI - Is sudden infant death syndrome a cause of death? PMID- 8418595 TI - Phenylalanine embryopathy in three siblings: implications of maternal diet therapy. PMID- 8418597 TI - Ought 'standard care' be the 'standard of care'? A study of the time to administration of antibiotics in children with meningitis. AB - OBJECTIVE: To determine the time from triage in an emergency department until administration of parenteral antibiotics in children with bacterial meningitis. RESEARCH DESIGN: Retrospective review of medical records and survey of medical subspecialists in infectious diseases and emergency medicine. SETTING: Emergency departments of two university-affiliated pediatric hospitals. PARTICIPANTS: All children with bacterial meningitis identified in medical records from 1987 to 1989 (N = 93). MEASUREMENTS: For each child, the time from presentation to the emergency department until administration of antibiotics (AB time) was determined; when possible, time from triage to contact with a physician, from triage to lumbar puncture, and from lumbar puncture to administration of antibiotics was measured. We then surveyed specialists in both pediatric infectious diseases (n = 23) and pediatric emergency medicine (n = 54) as to their beliefs about AB time in children with meningitis. STATISTICAL ANALYSES: Mann-Whitney Rank Sum Test and Kruskal-Wallis Test. RESULTS: Median AB time was 2.0 hours (interquartile range, 1.25 to 3.33 hours). Only one (1%) of 93 children received antibiotics within 30 minutes of presentation. Median time from triage until contact with a physician was 0.45 hour. Median time from lumbar puncture until antibiotics administration was about 0.5 hour. The estimates of median AB time differed significantly between emergency medicine (0.93 hour) and infectious disease (1.45 hours) experts, and estimates from both differed significantly from the median AB time (2.0 hours) actually observed. CONCLUSIONS: These data reveal that the usual and customary practice (ie, standard medical care) by qualified physicians may differ from opinions of standard medical care promulgated by medical experts. Even among experts there is a wide range of (mistaken) opinions about standard medical care. Insofar as jurors in medical malpractice cases are instructed to consider what physicians "ordinarily do in similar circumstances," a data-based definition of "standard" medical care should supplant anecdotal testimony by individual expert witnesses. PMID- 8418598 TI - Prognostic significance of oral lesions in children with perinatally acquired human immunodeficiency virus infection. AB - OBJECTIVE: To assess the prevalence and prognostic significance of oral lesions in children with human immunodeficiency virus infection. DESIGN: Clinical cohort study. SETTING: Children's Hospital Center at Jackson Memorial Hospital, Miami, Fla. PARTICIPANTS: Ninety-nine children with perinatally acquired human immunodeficiency virus infection examined longitudinally for oral lesions. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: Presence of oral lesions was determined by a pediatrician who had been trained in the diagnosis of oral lesions. The cumulative prevalence of lesions was 72% for oral candidiasis, 47% for parotid enlargement, and 24% for herpes simplex. The median time from birth to development of a lesion was 2.4 years for candidiasis, 4.6 years for parotid enlargement, and 5 years for herpes simplex. The median time from lesion to death was 3.4 years for patients with candidiasis, 5.4 years for patients with parotid enlargement, and 4.3 years for patients with herpes simplex. In a time-dependent proportional-hazards model, oral candidiasis was associated with a more rapid rate of progression to death (relative hazard, 14.2; 95% confidence interval, 4.8 to 41.8), while parotid enlargement was associated with a less rapid rate of progression to death (relative hazard, 0.38; 95% confidence interval, 0.16 to 0.88) and herpes simplex was unrelated to the rate of progression (relative hazard, 1.3; 95% confidence interval, 0.5 to 3.1). CONCLUSIONS: The presence of oral candidiasis and parotid enlargement confer important prognostic information and should be incorporated into decisions regarding therapy for HIV-infected children. PMID- 8418599 TI - Developing community faculty. Principles, practice, and evaluation. AB - Medical education is increasingly emphasizing outpatient experiences in community settings, and, concurrently, the development and assessment of the teaching skills of community faculty. These skills can be addressed through a faculty development program focusing on (1) issues relevant to community education, (2) adult learning principles, and (3) logistics that facilitate the participation of community medical faculty. Using a series of 1- to 3-day seminars, the program described focused on clinical precepting and brief presentations, used interactional teaching and practicums, and actively involved participants in the evaluation and planning fo the seminars. Evaluation of the program suggested that it was successful in increasing participants' knowledge and application of educational principles taught. Such programs can result in the development of an effective and potentially abundant resource of faculty members for the medical education of students and residents. PMID- 8418600 TI - Early intervention for children with or at risk of cerebral palsy. AB - Quantitative and qualitative analyses of well-conducted scientific studies of therapeutic early intervention for infants and young children with or at risk of a motor handicap failed to find convincing evidence for the efficacy of physical therapy. The shortcomings of present assessment tools are discussed and possible lines of enquiry identified. PMID- 8418601 TI - Dietary treatment and growth of hyperchylomicronemic children severely restricted in dietary fat. AB - OBJECTIVE: We followed the clinical course of four patients with type I hyperlipidemia from two kindreds who presented at an early age. PATIENTS: Two propositi presented with severe abdominal pain and bloody diarrhea at 8 and 10 weeks of age. They also exhibited delayed growth. We compared their course with that of two siblings (one sibling of each proband) who also have familial hyperchylomicronemia but were diagnosed and have subsequently shown normal growth. MAIN RESULTS: Although each sibling pair possesses the same lipoprotein lipase gene defect and resides in a similar environment, significant differences in stature are apparent. CONCLUSION: Specific structural defects in the lipoprotein lipase gene alone do not define phenotypic presentation. However, severity of clinical presentation may influence future growth characteristics. PMID- 8418602 TI - Use of zinc protoporphyrin measured by the Protofluor-Z hematofluorometer in screening children for elevated blood lead levels. AB - OBJECTIVE: To determine the usefulness of zinc protoporphyrin, as measured by the Helena Protofluor-Z hematofluorometer, for detecting elevated lead levels. DESIGN: Observational, descriptive review of laboratory records from a university toxicology laboratory. SETTING: Inner-city university pediatric clinic and two affiliated community clinics in Chicago, Ill. PATIENTS: Seven hundred seventy five children younger than 7 years with paired lead-zinc protoporphyrin results. MEASUREMENTS/RESULTS: Fifty-six percent had lead levels of at least 0.48 mumol/L and 8% had lead levels of at least 1.21 mumol/L. The sensitivity, positive predictive value, and negative predictive value of a zinc protoporphyrin level of 70 mumol/mol of hemoglobin for detecting a lead level of 0.48 mumol/L were 42%, 66%, and 50%, respectively, and for a lead level of 1.21 mumol/L were 74%, 18%, and 97%, respectively. Receiver operating characteristic curves demonstrated that for detecting lead levels of 0.48 mumol/L with zinc protoporphyrin, the probability of a true-positive result is close to that of a false-positive one. CONCLUSION: Zinc protoporphyrin is not a reliable screening test for detecting low blood lead levels. PMID- 8418603 TI - Radiological cases of the month. Bochdalek hernia after trauma. PMID- 8418604 TI - Radiological cases of the month. Septo-optic dysplasia (De Morsier syndrome). PMID- 8418605 TI - Picture of the month. Erythropoietic protoporphyria. PMID- 8418606 TI - Pathological cases of the month. Autosomal recessive polycystic kidney disease. PMID- 8418607 TI - Pathological cases of the month. Langerhans' cell histiocytosis with gastrointestinal involvement. PMID- 8418608 TI - Efficacy of cefixime in the treatment of acute otitis media in children. AB - OBJECTIVE: To compare the efficacy of cefixime with amoxicillin in the treatment of acute otitis media in children. DESIGN: Randomized, nonblinded study. SETTING: General pediatric clinic at a university hospital in Texas. PARTICIPANTS: A volunteer sample of 201 children, aged 2 months through 6 years. INTERVENTIONS: A 10-day oral course of cefixime (8 mg/kg per day administered once daily) or amoxicillin (40 mg/kg per day administered in three divided doses [every 8 hours]). MEASUREMENTS/MAIN RESULTS: Tympanocentesis for bacterial culture was performed on all affected ears on enrollment and after 4 to 6 days of therapy. The patients were evaluated clinically 4 to 6 days after starting therapy, at the end of therapy, and 3 to 4 weeks after therapy was completed. Using Fisher's Exact Test, no significant difference was found between the two treatment groups for rate of clinical improvement or rate of eradication of Haemophilus influenzae and Streptococcus pneumoniae. However, combining the results from this study and two previously reported studies, cefixime was found to be more effective in eradication of H influenzae and less effective in eradication of S pneumoniae. PMID- 8418609 TI - Fractures in young children. Distinguishing child abuse from unintentional injuries. AB - OBJECTIVE: To determine features of fractures in young children that would be helpful in distinguishing child abuse from unintentional injuries. DESIGN: Case series. SETTING: Pediatric Services of Yale-New Haven (Conn) Hospital (a tertiary care center). PATIENTS: Consecutive children who were less than 3 years of age and who were examined for a fracture from January 1979 through December 1983 were identified from the daily logs of the emergency department or the hospital's child abuse registry. OUTCOME MEASURE: Each case was rated, by means of predefined criteria and a consensus of two clinicians and two pediatric radiologists, on a seven-point scale from "definite child abuse" to "definite unintentional injury." A middle rating of "unknown" was used if there was not enough information to reach a consensus. RESULTS: Of the 253 fractures in 215 children that were identified, we categorized 24.2% as abuse, 8.4% as unknown, and 67.4% as unintentional injuries. Fractures that were considered likely due to abuse were (1) fractures in children whose caretakers reported either a change in the child's behavior, but no accidental event, or a minor fall, but the injury was more severe than expected; (2) fractures of the radius/ulna, tibia/fibula, or femur in children less than 1 year of age; or (3) midshaft or metaphyseal fractures of the humerus. Linear fractures of the parietal bone were the most common skull fractures, whether due to abuse or unintentional injuries. CONCLUSION: In young children with fractures, child abuse is common. By comparing fractures due to abuse and those due to unintentional injuries, we obtained empiric evidence to help clinicians and radiologists correctly examine children with such serious injuries. PMID- 8418610 TI - Hereditary pyropoikilocytosis. Clinical and laboratory analysis in eight infants and young children. AB - OBJECTIVE: To describe the mode of presentation and natural history of hereditary pyropoikilocytosis to promote earlier diagnosis and improve treatment of this disorder. DESIGN: Retrospective case analysis. SETTING: Tertiary care pediatric medical center. PARTICIPANTS: Eight children referred for examination. INTERVENTIONS: Treatment of symptomatic anemia and complications. MEASUREMENTS/MAIN RESULTS: Hereditary pyropoikilocytosis was responsible for a very characteristic perinatal course in our patient cohort. All patients had hyperbilirubinemia requiring either exchange transfusions or phototherapy. Peripheral blood smears at birth revealed nucleated red blood cells, marked poikilocytosis, microcytosis, and reticulocytosis. All other pertinent laboratory studies (eg, immune hemolysis, sepsis, hereditary spherocytosis, etc) were noncontributory. Specific red blood cell membrane analysis clearly identified these patients as having hereditary pyropoikilocytosis. Follow-up evaluation of these patients (in one instance spanning 12 years) determined that this disorder is associated with a clinically apparent anemia but an excellent prognosis. CONCLUSIONS: Red blood cell membrane analysis should be performed in neonates with a hemolytic anemia requiring therapy in which no identifiable cause can be ascertained with a conventional diagnostic examination. PMID- 8418611 TI - Growth hormone deficiency in two siblings with Alstrom syndrome. AB - OBJECTIVE: To investigate if there is an endocrinologic explanation for the short stature in patients with Alstrom syndrome. DESIGN: Patient reports. SETTING: The Children's Hospital of Philadelphia, Pa. PARTICIPANTS: Two siblings with Alstrom syndrome who were referred to the Endocrine Division for evaluation of obesity, growth, and glucose metabolism. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: The low growth velocity in both patients prompted growth hormone evaluation. The abnormal results of two provocative tests of growth hormone function, as well as low concentrations in frequent overnight sampling of serum growth hormone, were compatible with growth hormone deficiency. However, the patients had advanced bone ages, early normal growth, and normal insulinlike growth factor 1 concentrations. The results of the glucose tolerance tests revealed marked elevation of insulin (5955 pmol/L and 7677 pmol/L) and glucose intolerance consistent with insulin resistance. Pituitary stimulation studies revealed normal thyroid and gonadotropin axes. CONCLUSIONS: Growth hormone deficiency may account for the short stature in some patients with Alstrom syndrome. The advanced bone age and normal early growth may be due to hyperinsulinism. A specific defect in the signal transduction of insulin action and possibly hormone spillover on another receptor may account for the existence of insulin resistance in the presence of normal growth and growth hormone deficiency. PMID- 8418612 TI - The US Renal Data System and the Case-Mix Severity Study. AB - The US Renal Data System (USRDS), established in 1988 to collect and analyze information on end-stage renal disease (ESRD) patients in the United States and their treatment, is briefly described. A number of special studies have been undertaken by the USRDS. One of these is the Case-Mix Study, which reviewed data from an incident sample of 4,911 patients starting dialysis in 1986 and 1987 who were monitored for 1 to 4 years. Analysis included Cox proportional hazard modeling, which allowed quantification of the effect of various comorbid factors on mortality. Relevant findings included a greater mortality in diabetic patients treated by continuous ambulatory peritoneal dialysis (CAPD) compared with those treated by center hemodialysis. With regard to adequacy of dialysis, prescribed dialysis averaged a Kt/V of 1.0, with 53% of patients having a value of 1.0 or less. Where delivered Kt/V could be calculated, the mean of this was 0.72. The prescribed Kt/V in this sample of patients did not predict mortality, but there was a statistically significant inverse correlation of delivered Kt/V with mortality. It would appear a significant number of patients were being underdialyzed in the United States in 1986 and 1987. The means by which to obtain further information from the USRDS is described. PMID- 8418613 TI - Report of workshop on technique and technology. PMID- 8418614 TI - Epidemiology working group on morbidity and mortality of end-stage renal disease. PMID- 8418615 TI - Summary of the joint National Institutes of Health/Renal Physician's Association workshop on dialysis, morbidity and mortality. PMID- 8418616 TI - The current clinical database: is it complete and useful? PMID- 8418617 TI - Nutrition workshop. PMID- 8418618 TI - Basic science research. PMID- 8418619 TI - Pregnancy in women with end-stage renal disease: treatment of anemia and premature labor. AB - There is little experience with the use of various therapies in the end-stage renal disease patient who becomes pregnant. Erythropoietin for the treatment of anemia has become part of the standard treatment regimen of dialysis patients, but experience with its use in pregnancy is limited. We report five cases of its use in dialysis patients during pregnancy. We found no evidence that it crossed the placenta or that it made blood pressure control more difficult. We found that patients required a higher dose of erythropoietin to maintain hematocrit levels than they had before pregnancy. Another therapy involves the treatment for premature labor, which is the most common cause of pregnancy loss in dialysis patients. Two of our patients were successfully treated with indomethacin for premature labor. Both drugs are useful tools in the management of pregnant dialysis patients. PMID- 8418620 TI - End-stage renal disease in systemic lupus erythematosus. AB - The progression of lupus nephritis severe enough to require dialysis does not necessarily indicate that it is "end-stage." Ten percent to 28% of patients with lupus nephritis who develop renal failure requiring dialysis will recover enough function to come off dialysis. The clinical activity of systemic lupus erythematosus (SLE) is quiescent in most patients with end-stage lupus nephritis, regardless of the modality of dialysis treatment. Clinical and serologic remission of SLE permits judicious withdrawal of immunosuppressive therapy, as well as a favorable long-term outcome for patients that is comparable to that of nonlupus patients. The great majority of deaths in patients with end-stage lupus nephritis occur in the first 3 months of dialysis and most often result from infection. Later, infection and cardiovascular complications are common causes of death. Patients with lupus nephritis should wait at least 3 months on dialysis before receiving a kidney transplantation. Immunosuppressive therapy and graft survival rates for lupus patients are not different from those of nonlupus patients. Recurrence of lupus nephritis in the allograft is exceedingly rare. PMID- 8418621 TI - Isolation of human immunodeficiency virus from peritoneal dialysate. AB - Human immunodeficiency virus (HIV)-seropositive individuals represent a growing population of peritoneal dialysis (PD) patients. An important health care issue in these patients is the potential for their PD fluid to transmit virus. Some body fluids, such as urine, have been demonstrated to be negative for HIV and therefore presumably to be of low risk for virus transmission. To determine whether HIV could be recovered from PD fluid, we cultured the PD fluid from two asymptomatic HIV-seropositive patients and one patient with the acquired immunodeficiency syndrome (AIDS). HIV was isolated from both the PD fluid and the blood of two of the three patients tested, one being only HIV-seropositive and one with AIDS. These findings indicate that such fluid could potentially be a source of viral transmission and emphasize the need for conscientious application of universal precautions both in and out of the hospital. PMID- 8418622 TI - Serum albumin: a marker for morbidity in peritoneal dialysis patients. AB - This study was designed to determine if the serum albumin is a marker for morbidity or mortality in peritoneal dialysis (PD) patients. The impact of a low serum albumin on the risk of hospitalization, peritonitis, or death was examined in 71 patients. Blood urea nitrogen (BUN), cholesterol, age, and the presence or absence of diabetes were also examined. In independent analyses, the serum albumin was lower (32.7 +/- 5.6 v 36.3 +/- 4.3 g/L, P < 0.01), the diagnosis of diabetes was more frequent (41% v 7%, P < 0.01), and the number of episodes of peritonitis were greater (2.0 +/- 1.6 v 0.7 +/- 1.3, P < 0.01) in the group of patients hospitalized compared with those not hospitalized. When diabetics were excluded from analysis, the serum albumin remained significantly lower in hospitalized patients. Stepwise logistic regression analysis, excluding the 10 patients hospitalized only for treatment of peritonitis, confirmed that only a low serum albumin and the diagnosis of diabetes were independent predictors of increased morbidity as evident by the increased frequency of hospitalization. Every 10 g/L decrease in the serum albumin increased the odds ratio for hospitalization by 5.2. The diagnosis of diabetes resulted in a 10-fold increase in the odds ratio. We conclude that a low serum albumin serves as a marker of morbidity in PD patients, primarily as a marker of increased risk for hospitalization. The diagnosis of diabetes also greatly increases the likelihood of hospitalization. Peritonitis is a cause for hospitalization, but not an independent risk factor. PMID- 8418623 TI - Evaluation of platelet hyperfunction in hemodialysis patients receiving recombinant human erythropoietin. AB - There are data suggesting that recombinant human erythropoietin (rHuEPO) may induce thromboses in hemodialysis patients, possibly due to alterations in platelet function. In an earlier study, we found evidence of platelet hyperfunction in several patients 4 to 8 weeks following the start of rHuEPO therapy, which was begun shortly after hemodialysis was initiated. Studies were performed to examine the effects of rHuEPO on whole blood platelet aggregation and adenosine triphosphate (ATP) release independent of changes in hematocrit or the uremic state. Eight hemodialysis patients without and four with a history of vascular access clotting had platelet aggregation tests performed at baseline while receiving rHuEPO, off rHuEPO for 2 weeks, and 4 to 6 weeks after restarting the drug. While the plasma EPO level decreased significantly after the 2-week period off rHuEPO (P < 0.0001), the hematocrit did not change at any of the three time periods. Whole blood platelet aggregation in response to adenosine diphosphate (ADP), collagen, and ristocetin was not significantly altered on or off rHuEPO in either patient group. Platelet hyperfunction, determined by aggregation or ATP release either spontaneously or in response to low-dose ADP or ristocetin, was not seen in any patient. These data suggest that the increase in access clotting is not the result of platelet hyperfunction induced directly by rHuEPO. PMID- 8418624 TI - The use of tissue plasminogen activator to declot arteriovenous accesses in hemodialysis patients. AB - Thrombosis is the most common complication of arteriovenous (A-V) access, resulting in malfunction or total failure. We describe the first use of the thrombolytic agent tissue plasminogen activator (t-PA) to declot the A-V access in 15 hemodialysis patients (14 A-V grafts and one fistula). The t-PA was infused directly into the A-V access in 10-mg doses, at 2-hour intervals, to a maximum of 30 mg. As determined by angiography, t-PA infusion resulted in a dramatic decrease in clot volume in all cases and complete lysis, with return of bruit and thrill, in 10 patients. Eight of the 10 were able to be treated with hemodialysis via the A-V access the following day. In these patients, angiography demonstrated stenosis at the venous end of the A-V access in eight of nine A-V grafts (the one fistula did not have a venous stenosis). Three patients reclotted within 24 hours, and one had bleeding 5 days later after dialysis requiring compression of the A-V access, which resulted in reclotting. Five patients had functioning A-V grafts 1 to 15 months after t-PA treatment (with angioplasty of the venous stenosis required in three of these), and one patient was lost to follow-up. All five patients in whom t-PA infusion was only partially successful had venous stenosis. One patient died before surgery (unrelated to t-PA). Thus, venous stenosis was present in 13 of 15 A-V accesses studied, the highest incidence reported to date.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418625 TI - Acute aluminum toxicity and alum bladder irrigation in patients with renal failure. AB - Intravesical alum instillation is an increasingly common treatment for hemorrhagic cystitis. It has been claimed that this therapy is safe in patients with renal failure. We report the case of a patient with renal failure following a bone marrow transplantation who developed an acute encephalopathy from apparent aluminum intoxication following intravesical alum. In a review of the literature, we have found two similar cases of acute aluminum intoxication from alum in patients with renal failure who have received multiagent chemotherapy. We suggest that alternate therapies be selected for hemorrhagic cystitis in such patients. PMID- 8418626 TI - Immune complex glomerulonephritis with unusual microfibrillar deposits associated with primary bone marrow lymphoma. AB - Glomerular microfibrillary deposits are characteristic of several diseases of the kidney. In a number of glomerulopathies, the nature of these microfibrillary deposits is critical in classifying the renal lesion and in suggesting the possibility of an associated systemic process. However, it is likely that as efforts are made to classify glomerulopathies with microfibrillary deposits, certain cases will defy categorization. We describe one such case in which a patient presented with rapidly progressive glomerulonephritis associated with large subepithelial, parallel-arrayed microfibrillar deposits associated with a primary bone marrow B-cell lymphoma. While IgG, C3, and lambda and kappa light chains were deposited in the glomerulus, serum and urine protein electrophoresis were normal. Treatment with Cytoxan and prednisone caused simultaneous remission of the lymphoma and the glomerulonephritis. Relapse of the lymphoma was associated with rapid deterioration of renal function. This case may represent a newly described variant of immune complex-mediated glomerulonephritis associated with microfibrillary deposits. The possibility is raised that the glomerular lesion is due to atypical immunoglobulins synthesized by a bone marrow lymphoma. PMID- 8418627 TI - Malignant hypertension secondary to pheochromocytoma in a hemodialyzed patient. AB - Resistant hypertension and paroxystic hemodynamic changes are common features in patients on regular hemodialysis. We describe a hemodialyzed patient who developed malignant hypertension. The finding of elevated plasma catecholamines suggested the diagnosis of pheochromocytoma, which was confirmed by surgical removal of the tumor. PMID- 8418628 TI - Radiological and histological improvement of oxalate osteopathy after combined liver-kidney transplantation in primary hyperoxaluria type 1. AB - A 15-year-old patient with severe bone disease (with bilateral fractures of hips and shoulders) due to primary hyperoxaluria type 1 (PH1) was treated with combined liver-kidney transplantation after a 4-year hemodialysis period. Normalization of excessive oxalate synthesis brought in by the liver graft combined with the slow excretion of skeletal oxalate stores by the renal graft led to progressive improvement of clinical, radiological, and histological evidence of oxalate osteopathy. This allowed bilateral hip replacement 3 years after transplantation, which led to complete physical rehabilitation of the crippled patient. Combined liver-kidney transplantation constitutes the treatment of choice for end-stage renal failure due to PH1, even in the face of severe oxalate bone disease. PMID- 8418629 TI - Normal and abnormal nephrogenesis. AB - During the past decade, exciting advances in the fields of cell and molecular biology have provided new insight into the processes of normal and abnormal nephron induction and renal morphogenesis. Although the specific molecular signals that control renal mesenchymal-epithelium inductive interaction remain unknown, recent data suggest that postinductive nephrogenesis may be regulated by the overall balance of a number of local autocrine and/or paracrine growth factor systems. Alterations in the critical balance of regulatory factors might produce a variety of hypoplastic and dysplastic nephropathies or hyperplastic lesions such as tubular cysts. Additional studies demonstrate that extracellular matrix components and cell surface integrins have important regulatory roles in ureteric bud development and branching. Perturbations in matrix or integrin expression due to altered gene activity or toxin exposure would be expected to produce a variety of renal abnormalities ranging from failure of nephron induction (aplasia) to focal disruptions of differentiation (segmental dysplasia). Finally, several groups of genes encoding transcriptional regulatory proteins have been identified that appear to regulate aspects of cell proliferation, pattern formation, and segment-specific differentiation during normal and abnormal nephrogenesis. Future studies will elucidate the roles that specific genes and proteins play in renal development and will ultimately reveal the manner in which their dysregulation or dysfunction causes a variety of developmental renal disorders. PMID- 8418630 TI - Cell adhesion molecules in inflammation and thrombosis: status and prospects. AB - Cell-cell and cell-matrix interactions play vital roles in morphogenesis, inflammation, thrombosis, wound healing, immune surveillance, and growth and metastasis. A number of cell surface receptors, so-called cell adhesion molecules (CAMs), mediate these events in various cell types. In leukocytes and platelets, several CAM families interact with each other or with the extracellular matrix in a well-orchestrated manner, to bring these circulating cells to areas of antigenic challenge and tissue injury. Significant progress has been made recently in understanding the molecular basis of leukocyte recruitment into tissues and the formation of the hemostatic plug. Several murine or humanized monoclonal antibodies directed against key adhesion receptors that mediate these events have been found effective in taming immune and thrombotic injury in experimental animal models and in phase I clinical trials. These advances herald the arrival of a novel class of antiinflammatory and antithrombotic drugs, the antiadhesions, which will likely be effective in many clinical situations facing the nephrologist and internist. PMID- 8418631 TI - Cardiac and cerebrovascular disease in chronic uremia. AB - Cardiovascular disease causes death in more than 40% of dialysis patients and the burden of its morbidity is high. The relationships between traditional risk factors for cardiac mortality, as identified by the Framingham study, and potential uremia-related risk factors are unclear. The characteristic echocardiographic pattern in dialysis patients is a dilated left ventricle with normal systolic function and left ventricular hypertrophy. Dilated cardiomyopathy, severe left ventricular hypertrophy, and coronary artery disease occur frequently and predispose to congestive heart failure, arrhythmias, and ischemic heart disease. The risk factors that predispose to each disease and the natural history of each disorder have not been studied with sufficient precision. Few quality studies have been undertaken to identify the interventions that effectively ameliorate the impact of cardiac disease in dialysis patients. In some subsets of patients, erythropoietin or renal transplantation will be useful in inducing regression of some cardiac diseases. However, the impact of various antihypertensive agents, lipid-lowering agents, or normalization of hematocrit is unknown. PMID- 8418632 TI - Chronic renal failure is a state of cellular calcium toxicity. AB - Studies on the metabolic profile of many cells have shown that chronic renal failure (CRF) is associated with a significant elevation in the basal levels of cytosolic calcium ([Ca2+]i). This latter abnormality is, in major part, responsible for the organ dysfunction in CRF. The initial step leading to the eventual increase in the basal level of [Ca2+]i is parathyroid hormone (PTH) mediated increased calcium influx into cells. This is followed by decreased extrusion of calcium out of cells due to reduced activity of the enzymes responsible for pumping calcium out of the cells. The combination of increased entry and decreased exit of calcium results in elevation of [Ca2+]i. Prevention of secondary hyperparathyroidism in CRF or blocking of the effect of PTH by a calcium channel blocker results in normalization of [Ca2+]i and restoration of cell function. Thus, the available data are consistent with the notion that CRF is a state of cellular calcium toxicity, which underlies many of the metabolic and functional derangements in CRF. PMID- 8418633 TI - Lipoprotein metabolism in normal and uremic patients. AB - Abnormalities in circulating lipoprotein concentrations are a characteristic finding both in patients with uremia and in patients undergoing dialysis. These patients tend to have elevated triglyceride (TG) concentrations and low concentrations of high-density lipoprotein (HDL) cholesterol. Elevations of low density lipoprotein (LDL) are not usually observed unless the patients have undergone renal transplantation and are receiving therapy with immune suppressive medications. Hypertriglyceridemia and low HDL may be the consequence of decreased actions of lipoprotein lipase (LPL), the endothelial cell-bound enzyme that degrades circulating lipoprotein triglyceride. A poorly characterized circulating inhibitor to this enzyme is found in uremic plasma. Preliminary data suggest that high-flux dialysis with polysulfone (PS) membranes improves the lipoprotein abnormalities and decreases circulating LPL inhibitors. Whether such therapy will alter the incidence of coronary morbidity and mortality in patients with end stage renal failure remains to be tested. PMID- 8418634 TI - Tapering or discontinuing cyclosporine for financial reasons--a single-center experience. AB - In patients with primary cadaveric renal transplants and stable allograft function, we assessed the impact of tapering or discontinuing cyclosporine A (CsA) for financial reasons. Forty-two patients whose CsA was discontinued ("no dose") and 29 patients whose CsA was tapered to 100 to 150 mg/d ("low-dose"; mean, 1.7 mg/kg/d) were examined. Results were compared with 70 age- and race matched control patients maintained on at least 200 mg/d of CsA (mean, 3.9 mg/kg/d). Follow-up time for all patients averaged 55 +/- 18 months. Late acute rejection episodes occurred more frequently in no-dose than in low-dose (P = 0.017) or control (P = 0.001) patients. In the no-dose group, blacks experienced a greater number of late acute rejections than whites. These late acute rejections often coincided with the discontinuation of CsA and contributed to an increased rate of allograft loss in blacks in the no-dose group compared with black and white controls (P = 0.011). In contrast, no increase in late acute rejection episodes occurred in blacks tapered to low doses of CsA. Black patients who remained on low doses of CsA also exhibited a trend toward allograft survival that was intermediate between that of control and no-dose patients. In those patients who retained functional allografts, mean serum creatinine concentration did not differ between the study groups at the beginning and end of the follow-up period. These findings support continuance of CsA in black primary cadaveric renal transplant patients, even if dosages must be reduced to 100 to 150 mg/d.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418635 TI - Mechanisms that cause protein and amino acid catabolism in uremia. AB - Anorexia and/or a protein- and calorie-restricted diet can cause protein wasting by limiting the intake of essential amino acids (EAA) and, hence, protein synthesis. By this mechanism plus the effects of inadequate calories, restricted diets could contribute to the loss of lean body mass of uremic patients. Uremia also impairs the normal metabolic responses that must be activated to preserve body protein, thereby augmenting the adverse effects of anorexia. The responses impaired are those that conserve EAA and protein, which results in catabolism of EAA and muscle protein. An important factor that initiates abnormal adaptive responses in uremia is metabolic acidosis, because acidosis stimulates muscle protein degradation and increases the activity of branched-chain ketoacid dehydrogenase and, hence, the catabolism of branched-chain amino acids (BCAA). The effects of acidosis could be mediated by impaired regulation of intracellular pH and/or an increase in glucocorticoid production. Research directed at identifying the specific proteolytic pathways that are activated by metabolic acidosis has excluded a major role for Ca(2+)-activated or lysosomal proteases and suggests activation of an adenosine triphosphate (ATP)- and ubiquitin dependent proteolytic pathway. The mechanism of activation of this pathway includes an increase in mRNA for enzymes involved in protein and amino acid catabolism. PMID- 8418636 TI - Adequacy of peritoneal dialysis. AB - The combined annual mortality and drop out rate for peritoneal dialysis (PD) patients is relatively uniform worldwide at approximately 35%. The level of PD therapy prescribed in clinical practice is largely empirical and typically consists of four 2-L exchanges daily. It might be speculated that the 35% annual attrition rate in PD may in part reflect under dialysis in some patients due to empirical rather than quantitative and individualized prescription of PD therapy. Urea kinetic modeling has been successfully used to quantitatively prescribe hemodialysis (HD) therapy and, in principle, it should be able to serve the same purpose in PD. Comparison of HD and PD is complicated, because peritoneal urea clearance is virtually continuous, while in HD clearance is provided only about 5% of the time and urea accumulates over 95% of each treatment cycle. The blood urea nitrogen (BUN) in PD (BUNpd) is essentially constant and reflects the steady state, while in HD a sawtooth BUN profile results that reflects the short intermittent dialyses. The HD BUN profile can be characterized by either the predialysis level (BUNo) or the time-averaged concentration (TAC) over each treatment cycle. TAC is substantially lower than BUNo due to the sharp obligatory decrease in BUN during each short high-clearance dialysis. The rate of clearance required in HD is approximately 30 times higher than in PD, and the total clearance (KT) required in HD is 50% higher than in PD to achieve BUNpd = BUNo (at identical normalized protein catabolic rate [NPCR]), which reflects the decreasing urea flux rate during HD due to the decreasing BUN.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418637 TI - Nutrition in hemodialysis patients. AB - Malnutrition has been recognized in maintenance hemodialysis patients since the initiation of this long-term therapy. The cause of protein-caloric malnutrition in the chronic dialysis population is multifactoral, with decreased intake, increased loss, and altered metabolism as likely major categories. Despite marked improvement in the techniques and adequacy of hemodialysis and peritoneal dialysis, protein and caloric therapy has not changed in dialysis patients. Assessment of malnutrition has not been pursued as vigorously as necessary. Moreover, failure to document the diagnosis has distorted interpretation of the reported increasing mortality in this population. More intensive assessment and documentation of malnutrition states, along with alteration in the therapeutic approach, is warranted if we hope to positively affect the mortality of maintenance dialysis patients. PMID- 8418638 TI - Hyperphalangism, facial anomalies, hallux valgus, and bronchomalacia: a new syndrome? AB - We report on a boy with hyperphalangism, partial syndactyly, facial anomalies, and diffuse bronchomalacia, born to a nonconsanguineous French-Canadian couple. To our knowledge, this is a hitherto undescribed syndrome. PMID- 8418639 TI - Absent chondrodysplasia punctata in a male with an Xp terminal deletion involving the putative region for CDPX1 locus. AB - This is a follow-up report on a male patient with a 46,Y,r(X) karyotype. Although he had no clinico-radiological features of X-linked recessive chondrodysplasia punctata (CDPX1), molecular studies revealed an Xp terminal deletion involving the putative region for the CDPX1 locus (PABX-DXS31). We suspect that the absence of CDPX1 may be attributable to the nature of the disease and the extreme short stature of the patient (mean -5.6 S.D.). PMID- 8418640 TI - Rare variant of chromosome 9. PMID- 8418641 TI - Genetic consequences of "euchromatic" band within 9qh region. PMID- 8418642 TI - Is loss of band 7p21 really critical for manifestation of craniosynostosis in 7p ? PMID- 8418643 TI - X-linked familial exudative vitreoretinopathy (FEVR): results of DNA analysis with candidate genes. PMID- 8418644 TI - Low level of mosaicism for a balanced translocation as a case of unbalanced offspring. PMID- 8418645 TI - Annular pancreas in a mother and daughter. PMID- 8418646 TI - Transmission of Proteus syndrome from mother to son? PMID- 8418647 TI - Gonadal function in Smith-Lemli-Opitz syndrome. PMID- 8418648 TI - Hypomelanosis of Ito with trisomy 18 mosaicism. PMID- 8418649 TI - Dolichomegasigmoid in Aarskog syndrome. PMID- 8418650 TI - New diagnostic method for Pallister-Killian syndrome: detection of i(12p) in interphase nuclei of buccal mucosa by fluorescence in situ hybridization. AB - Detection of the supernumerary isochromosome 12p [i(12p)] was performed on buccal smear preparations from 2 patients with Pallister-Killian syndrome, 21 (patient 1) and 15 months (patient 2) old, by interphase fluorescence in situ hybridization (FISH) using a chromosome 12-specific alpha satellite probe. Isochromosome 12p-positive cells were identified by observing 3 signals over the nucleus, while diploid cells had 2 signals. The proportion of i(12p)-positive cells thus identified was high in the epithelial cells of buccal mucosa at 68 and 53% from patients 1 and 2, respectively. Further, the frequencies of i(12p) positive cells were also studied in PHA-stimulated peripheral lymphocytes, cultured skin fibroblasts (both patients), and directly harvested T and B-cells (patient 1). Of these tissues, buccal mucosa showed the highest proportion of i(12p)-positive cells. These findings indicate that epithelial cells of buccal mucosa are likely to retain i(12p)-positive cells. Detection of i(12p) using direct buccal smear preparations by interphase FISH is a rapid, effective and non invasive method for confirming the diagnosis of the Pallister-Killian syndrome. PMID- 8418651 TI - Duplication (20p) in association with thyroid carcinoma. AB - We report on a girl with short stature, mental retardation, mutism, "coarse" facial appearance, and papillary-follicular thyroid carcinoma. She had dup(20p) derived from a paternal balanced translocation [(12p;20p)]. We speculate that the carcinoma in our patient may be related to the deletion of material from 12p resulting in absence of genetic material normally required for the suppression of thyroid tumorigenesis. PMID- 8418652 TI - Fetal and neonatal outcome of exposure to anticoagulants during pregnancy. AB - We studied fetal and neonatal outcome of women maintained on anticoagulants (warfarin and/or heparin) during pregnancy. Among 22 Chinese families, 13 mothers (59%) had a history of recurrent abortion or stillbirth while being maintained on warfarin treatment. Twenty-nine liveborn children (17 boys, 12 girls), ages 0.6 11.3 years at follow-up, were analysed for evidence of embryopathy. These were subdivided into 2 groups. Group 1 consisted of 18 children (12 boys, 6 girls) whose mothers were only given warfarin during pregnancy. Five were small for gestational age, and 12 had features of warfarin embryopathy such as nasal hypoplasia. One had subependymal intraventricular hemorrhage shown on neonatal ultrasonography. Group 2 consisted of 11 children (5 boys, 6 girls) whose mothers were maintained on warfarin and heparin during pregnancy. Three were premature deliveries, and 4 had nasal hypoplasia. One had cleft lip, cleft palate, cataract, microphthalmia, intraventricular hemorrhage, and hydrocephalus. We found that despite the high risk of fetal wastage, there was a relative lower risk of major complications, except for some minor cosmetic defects such as nasal hypoplasia. This might lead to readjustment of advice concerning contraception given to pregnant women who were maintained on anticoagulant therapy. PMID- 8418653 TI - Characterization of a de novo 48,XX,+r(X),+r(17) by in situ hybridization in a patient with neurofibromatosis (NF1). AB - We describe a patient with familial neurofibromatosis (NF1), short stature, developmental delay, and a de novo chromosome abnormality. In situ hybridization was done using chromosome specific centromere probes to characterize the karyotype as 46,XX/47, XX,+r(X) (p11q11)/47,XX,+r(17) (p11q11)/48, XX,+r(X) (p11q11),+r(17) (p11q11). The NF1 mutation, as well as each supernumerary ring chromosome, may have played a role in perturbing the normal developmental process of this patient. PMID- 8418654 TI - Quantitation of craniofacial anomalies in utero: fetal alcohol and Crouzon syndromes and thanatophoric dysplasia. AB - The study of fetal growth and development by ultrasound has been greatly facilitated in the past few years by the availability of anthropometric standards for the fetal body. Thus, the obstetrician is able to discern between normal and grossly abnormal, and even to quantitate certain fine fetal structures such as the face. This paper presents results obtained from a group of 5 patients referred to the Medical Center from private practices in Indianapolis, Indiana. Prenatal cephalometric analyses by ultrasound suggested the presence of craniofacial anomalies in all 5 cases. However, such defects were not detectable by routine ultrasonographic examination. A clinical examination after birth of each of these 5 patients suggested the following diagnoses: Fetal Alcohol Syndrome (FAS) in 2 individuals, Fetal Alcohol Effects (FAE) in one individual, Crouzon Syndrome (CS) in one patient, and Thanatophoric Dysplasia (TD) in one patient. In order to compare the craniofacial measurement values for each patient to normal standards, we developed Z-Score profiles and Pattern Variability Indexes (PVI) as described by Garn et al. [1984, 1985]. The values presented here support the idea that even mildly abnormal fetal craniofacial patterns are detectable by this relatively new application of ultrasound. At the present time, no conclusions can be made regarding the diagnostic accuracy of these patterns and profiles. However, the potential value of fetal cephalometry for documenting craniofacial dysmorphology is clearly indicated. PMID- 8418655 TI - Osteoporosis-pseudoglioma syndrome. AB - Two patients with osteoporosis pseudoglioma syndrome are described. Both are single children, born to nonconsanguineous, healthy parents. The first patient, a 17-year-old girl, had serious visual impairment since birth. She is severely dwarfed and has major skeletal deformities resulting in inability to walk since age 2 years. The second patient is an 18-year-old girl with unilateral neonatal blindness, short stature and deformities, mainly of pelvis and lower limbs. She has been able to walk with support up to now. The clinical and radiological findings in these 2 patients reflect the clinical variability of the condition. Results of collagen studies in both patients are normal and differentiate this condition clearly from severe osteogenesis imperfecta, which it resembles. PMID- 8418656 TI - Apparent Greig cephalopolysyndactyly and sinus node disease. AB - We present the clinical findings and follow-up data from birth to 10.5 years in a boy with Greig cephalopolysyndactyly who, in addition, presents sinus node disease ("sick sinus syndrome"). The significance of the concurrence of Greig cephalopolysyndactyly syndrome, an autosomal dominant condition mapped at 7p13, and sinus node disease is discussed. PMID- 8418657 TI - Reluctance to undergo predictive testing: the case of Huntington disease. AB - The development of a presymptomatic test for Huntington Disease (HD) has enabled some persons at risk to determine whether or not they are gene carriers. The purpose of this study was to explore the reasons why those at risk choose not to be tested in a situation where testing is available and most of the test associated costs are covered by state funding. Subjects were also asked their levels of knowledge about testing, attitudes towards aspects of the testing protocols, and intentions towards testing once the gene for HD is found. Sixty six individuals at risk for HD who had chosen not to be tested completed a mailed questionnaire. The most important reasons for not being tested were increased risk to children if one was found to be a gene carrier, absence of an effective cure, potential loss of health insurance, financial costs of testing, and the inability to "undo" the knowledge. Individuals comprising this sample were quite knowledgeable about predictive testing. Most supported the availability of testing despite the lack of a cure, the need for special counseling prior to testing, and the idea that testing should be a voluntary decision. Most said they would take the test if a treatment was available, if the mechanics of the test were simplified, or if the test was 100% accurate. The risk to relatives, lack of treatment or cure, fear of losing one's health insurance, and the accuracy of the information to be gained from testing are important factors in the decision not to be tested. PMID- 8418658 TI - Familial t(11;13)(q21;q14) and the duplication 11q, 13q phenotype. AB - Cases of duplication of distal 11q or proximal 13q have been reported independently. A specific translocation resulting in duplication of distal 11q, [der(22)t(11;22)(q23;q11)], has been documented in over 40 cases. We report on a male fetus with chromosomal excess of both distal 11q and proximal 13q resulting from a familial translocation. This case supports the causal association of duplication 11q with neural tube defects. PMID- 8418659 TI - Interstitial deletion of chromosome 2 region in a malformed infant. AB - We describe an infant with a lumbar meningomyelocele and other congenital anomalies and a de novo deletion of 2q36 with a non-mosaic karyotype 46,XX,del(2)(q36). PMID- 8418660 TI - Pericentric inversion of chromosome 4 giving rise to dup(4p) and dup(4q) recombinants within a single kindred. AB - Theoretically, every pericentric inversion can give rise, during meiosis, to 2 alternate recombinant chromosomes. One of these will have a duplication of short arm material and deletion of long arm material (dup p), and the other, a duplication of a long arm material and deletion of short arm material (dup q). However, most published cases have been limited to a single recombinant type occurring within a given kindred. Here we document a large pericentric inversion of chromosome 4 which gave rise, within 2 generations of a kindred, to both dup p and dup q recombinants. The family was ascertained by the birth of a baby girl with multiple congenital anomalies suggestive of Wolf-Hirschhorn syndrome, and was found to have a dup 4q recombinant. Subsequent studies of her father and of her 27-year-old mentally retarded aunt showed a balanced inv(4) (p15.32q35) and a dup 4p recombinant, respectively. Given that: (a) the balanced inversion involves approximately 87% of the length of chromosome 4; (b) the predicted meiotic pairing would be homosynapsis with loop formation; (c) the size of the segments distal to the breakpoints of the inversion are of similar and relatively small size; and (d) both recombinants are compatible with life, then the risk for recurrence of a recombinant in this family is high. Genetic counseling addressed these issues, and to date, both chronic villus sampling (CVS) and amniocentesis have been provided for prenatal diagnosis. PMID- 8418661 TI - Preliminary definition of a "critical region" of chromosome 13 in q32: report of 14 cases with 13q deletions and review of the literature. AB - We report on 14 patients with partial deletions of chromosome 13q. These patients exhibit a wide spectrum of phenotypes. Deletions limited to proximal bands q13 q31 are associated with growth retardation but not with major malformations. We review the literature since 1975 and summarize 13q deletion cases which have a phenotype involving one or more major malformations and mental retardation. Analysis of the breakpoints of these cases, as well as those reported by us, supports the hypothesis that only deletions involving at least part of band q32 are associated with major malformations and digital abnormalities. Patients with more distal deletions have severe mental retardation but do not have major malformations or growth retardation. A group of patients in whom the breakpoint is stated to be within q32 has had an intermediate phenotype. This suggests that it may be possible to define subregions within q32 whose deletion is associated with particular developmental defects. PMID- 8418662 TI - Congenital defects of lower limbs and associated malformations: a population based study. AB - As part of a detailed study of limb defects and associated patterns of congenital malformations, cases with lower limb deficiencies were analysed separately. We identified a total of 130 cases with deficiencies of the lower limbs without defects of the upper limbs. This gives an incidence of 1.07/10,000 livebirths, or 1/9,337 for this group of limb defects. Most common were femur deficiencies and deficiencies of the foot. A preponderance of males was found in the group of transverse defects of the leg (fibula/tibia deficiencies) and central axis deficiencies, while females had significantly more often longitudinal tibia defects and preaxial ray defects. PMID- 8418663 TI - Balanced complex rearrangement involving chromosomes 8, 9, and 12 in a normal mother, derivative chromosome 9 with recombinant chromosome 12 in her daughter with minor anomalies. AB - We report on a 19-month-old girl with a derivative chromosome 9 and a recombinant chromosome 12 resulting from a maternal balanced complex rearrangement involving chromosomes 8, 9, and 12. The karyotype of the phenotypically normal mother was 46,XX,t(8;12) (9;12) (8qter-->8p23::12q12-->12q 15::9q32-->9qter;9pter- >9q32::12q15--> 12qter; 12pter-->12q12::8p23-->8pter). The child's karyotype was 46,XX,-9,-12, +der(9) (9pter-->9q32::12q15-->12qter), +rec(12) (12pter- >12q15::9q32-->9qter) mat. The child had severe growth retardation, minor anomalies including trigonocephaly, hypertelorism, broad nasal root, apparently low-set and posteriorly angulated ears, triangular face, pectus carinatum, clinodactyly of fifth fingers, and almost normal psychomotor development. To the best of our knowledge, there have been only 3 previous reports of recombination derived from parental complex chromosome rearrangements. In the recombination products, the chromosomes were apparently balanced and the offspring had no clinical abnormalities. The present case exhibited abnormalities and may have a submicroscopic aberration of 12q arising from crossing over during maternal meiotic pairing, although her chromosomes appeared to be balanced. PMID- 8418664 TI - Potential role of an additive genetic component in the cause of amyotrophic lateral sclerosis and parkinsonism-dementia in the western Pacific. AB - Amyotrophic lateral sclerosis (ALS) and parkinsonism-dementia (PD) are neurological degenerative disorders that occur in three high incidence foci in the western Pacific: among the Chamorros of Guam and the Commonwealth of the Northern Marianas Islands, among Japanese on the Kii peninsula of Honshu Island, and among the Auyu and Jakai peoples of southern West New Guinea. Previous studies have implicated both genetic susceptibility and environmental risk factors in the causation and familial clustering of these disorders. The data analyzed consist of 2,026 individuals in nuclear families ascertained on Guam through two mechanisms: (1) nuclear families were included in the study if one or both parents in the family were affected with ALS or PD or both; and (2) a group of "controls" was selected by obtaining nuclear families where neither parent was affected and both had lived through the age of risk. Clinically, ALS and PD are two distinct disorders. However, preliminary analyses indicated that combining all three diagnoses into one affected diagnosis for genetic analyses (thereby assuming any genetic effect on susceptibility to the two disorders was due to the same genetic mechanism) was reasonable. An age, sex and birth cohort-specific liability was defined and segregation analysis was performed under both logistic and normal models for this liability at the time of disease onset. Under either model, purely environmental, Mendelian dominant and Mendelian recessive hypotheses could be rejected, but a two-allele additive major locus hypothesis could not be rejected. PMID- 8418665 TI - Cutaneous malignant melanoma and atypical moles associated with a constitutional rearrangement of chromosomes 5 and 9. AB - Hereditary cutaneous malignant melanoma (HCMM) with or without atypical moles has been described in several large kindreds worldwide. Despite numerous studies of these kindreds, the gene or genes responsible for this disorder has not yet been identified. Cytogenetic and molecular studies of melanoma tumor tissue and cell lines suggest that a gene involved in the pathogenesis of malignant melanoma lies on chromosome 9p. We describe a woman with atypical moles and multiple primary melanomas, who lacks a family history of HCMM, and, has a de novo constitutional unbalanced reciprocal translocation involving the short arms of chromosomes 5 and 9 with a cytogenetically visible deletion of 5p or 9p: [46,XX,-5,-9, +der(5)t(5;9) (p13.3 or 14.2;p13.3 or 21.2), +der(9)t(5;9)]. This finding supports the hypothesis that a gene predisposing to HCMM lies on the short arm of chromosome 9. PMID- 8418666 TI - National Neurofibromatosis Foundation International Database. AB - The National Neurofibromatosis Foundation International Database is a system for collecting comprehensive information on the clinical manifestations and natural history of neurofibromatosis. Data are entered into personal computers at participating centres and are pooled at the Central Database and coordinating centre in Vancouver, Canada. The system includes special mechanisms to assure consistency among centres and to maintain patient confidentiality. The database is designed to foster collaborative clinical and molecular genetic research in many aspects of neurofibromatosis. PMID- 8418667 TI - Phenotype of cranioectodermal dysplasia with different hair and bone abnormalities. AB - We report on a 3-year-old boy with hair abnormalities and a generalized bone dysplasia. He had very short, sparse hair and craniosynostosis. His stature, growth, and limb lengths were normal, as was his neurological development. While this phenotype has some resemblance to cranioectodermal dysplasia, the radiographic and hair abnormalities are different. Histological studies showed abnormalities in the internal root sheath of the hair follicle and the hair shaft. These findings define a new ectodermal dysplasia syndrome of unknown cause. PMID- 8418668 TI - Stable inheritance of the CMT1A DNA duplication in two patients with CMT1 and NF1. AB - Charcot-Marie-Tooth disease type 1A (CMT1A) was recently demonstrated to be associated with a large DNA duplication in 17p11.2p12. The gene for neurofibromatosis type 1 (NF1) or von Recklinghausen disease maps to 17q11.2. We have identified 2 unrelated patients who were diagnosed with both CMT1 and NF1. Molecular analysis of these patients demonstrated the presence of the CMT1A duplication and inheritance of this DNA rearrangement from a parent affected with CMT. Analysis of genomic DNA isolated from the neurofibroma removed from one of these patients showed the same 500 kb SacII junction fragment associated with the CMT1A duplication that was found in genomic DNA isolated from the blood. These results lend further support to the hypothesis that the CMT1A duplication is a stable DNA rearrangement. In addition, the molecular analysis of these 2 patients suggests that 2 common autosomal dominant conditions (CMT1 and NF1) can occur in the same individual, not because of an underlying single molecular defect, but rather, secondary to a chance phenomenon. PMID- 8418669 TI - Interstitial deletion of distal chromosome 4p in a patient without classical Wolf Hirschhorn syndrome. AB - We report on a patient with a de novo interstitial deletion of chromosome 4p; 46,XY,del(4) (p15.31p16.3). The cytogenetic diagnosis would predict a patient with the Wolf-Hirschhorn syndrome (WHS) since deletions of 4p16 are associated with WHS [Wilson et al., 1981]. This patient lacks the facial characteristics of WHS, but has some anomalies of WHS that are also commonly seen in other syndromes, i.e., severe growth retardation, developmental delay, and hypospadias. His molecular distal breakpoint occurs in 4p16.3 as defined by fluorescence in situ hybridization and Southern blot analysis, and his deletion does not overlap with the currently proposed WHS critical region. This case gives further support to the distal position of the WHS critical region and demonstrates some of the WHS associated phenotypes that can be attributed to a deletion of the proximal third of 4p16.3. PMID- 8418670 TI - 13th Annual Meeting of the Society of Perinatal Obstetricians. San Francisco, California, February 8-13, 1993. Abstract. PMID- 8418671 TI - Reliability analysis in therapeutic research: practice and procedures. AB - Twenty studies examining the reliability of assessment devices and outcome measures in therapeutic research were reviewed and analyzed. The 20 investigations contained 215 quantitative reliability values published in either the American Journal of Occupational Therapy or Physical Therapy during the past 5 years. The reliability studies were classified as interrater, intrarater, test retest, or internal consistency. Examination of interrater reliability accounted for 41% of all reported reliability values. Studies published in Physical Therapy were more likely to be concerned with test-retest reliability, whereas studies published in the American Journal of Occupational Therapy more often focused on interrater reliability. Examination of the data revealed that the intraclass correlation coefficient (ICC) was the most frequently reported estimate of reliability, accounting for 57% of all reported reliability coefficients. Further review of the results indicated that Pearson product-moment correlations and percentage of agreement indexes accounted for 22% of all reliability values reported in the studies examined. The Pearson product-moment correlation measures association or covariation among variables, but not agreement, and percentage agreement indexes do not correct for chance agreement. The argument is made that product-moment correlations and percentage agreement indexes are inadequate measures of interrater, intrarater or test-retest agreement. They should be used and interpreted with caution. PMID- 8418672 TI - The use of a game to promote arm reach in persons with traumatic brain injury. AB - This study tested a principle of occupational therapy and motor learning theory in the context of neurodevelopmental treatment techniques. Ten trials of occupationally embedded intervention (playing Simon, a computer-controlled game) were compared with 10 trials of rote arm-reach exercise. A counterbalanced design was structured so that each subject experienced each condition one week apart. Subjects were 17 men and 3 women with traumatic brain injury who exhibited mild to moderate spasticity in the upper extremity. Maximum distance from hip to wrist during active reach of the affected extremity was measured by digitization of videotape with the Motion Analysis EV-3D system. Results indicated that the use of the game elicited significantly more range of motion than the rote exercise (t (19) = 5.77, p < .001). These results support the use of an occupationally embedded intervention for persons with traumatic brain injury and add to the theoretical base of occupational therapy. PMID- 8418673 TI - An analysis of computer-related articles in occupational therapy periodicals. AB - The introduction of personal computers in the late 1970s and early 1980s has affected occupational therapy. Nine journals, three trade newspapers, and one series book published over an 11-year period by four English-speaking countries were catalogued for articles pertaining to computer use in occupational therapy. This investigation yielded 174 articles that were categorized by information type, application of computers covered, sophistication of technology, computer brand, and country of publication. Most articles provided general information, described clinical training applications, used an Apple computer, and were from an American publication. Few articles were reports of research. The percentage of computer-related articles has substantially increased in the observed 11-year period. These findings indicate increased computer use in occupational therapy. PMID- 8418674 TI - Increasing medication compliance in a woman with anoxic brain damage and partial epilepsy. PMID- 8418675 TI - Women's mental health: implications for occupational therapy. AB - This article describes issues of sexism in the mental health profession. Theoretical bias in psychiatry and the sociopolitical status of women are both linked to the inappropriate mental health treatment of women. A relational model of women's psychological development is offered as an alternative theoretical perspective for understanding the mental health needs of women. Ways to integrate this perspective in occupational therapy practice are included. PMID- 8418676 TI - A hierarchical model for evaluation and treatment of visual perceptual dysfunction in adult acquired brain injury, Part 1. AB - A developmental framework for evaluation and treatment of visual perceptual deficits in adults with acquired brain injury is presented. The framework is based on a review of research conducted on post-cerebrovascular accident subjects and subjects with traumatic brain injuries. Visual perceptual skill is conceptualized as a hierarchy of skill levels that interact and subserve each other. Higher level skills evolve from integration of lower level skills and are subsequently affected by disruption of lower level skills. Oculomotor control, visual field, and acuity form the foundation skills, followed by visual attention, scanning, pattern recognition, memory, and visual cognition. Brain injury can affect the integrity and interaction of each skill level and affect daily living function. Application of this framework dictates a bottom-up approach to evaluation and treatment, emphasizing identification and remediation of deficits in lower level skills to allow normal integration of higher level skill. PMID- 8418677 TI - A hierarchical model for evaluation and treatment of visual perceptual dysfunction in adult acquired brain injury, Part 2. AB - A framework for evaluation and treatment of visual perceptual dysfunction in adults with acquired brain injury is presented. The framework is based on the concept of a hierarchical structure of perceptual skill levels that interact and subserve one another. Higher level skills in the structure evolve from the integration of lower level skills and are subsequently affected by disruption of the lower level skills. Oculomotor control, the visual fields, and visual acuity form the foundation skills in the framework, followed by visual attention, scanning, pattern recognition, memory, and visual cognition. The order of evaluation and treatment is dictated by the framework. Emphasis is placed on identification and remediation of deficits in the lower level skills that will cause spontaneous improvement of higher level skills. Three treatment principles and five training guidelines are presented that reflect this concept. Specific examples of treatment tasks are provided. PMID- 8418678 TI - Level I fieldwork: creating a positive experience. AB - Qualitative research methodology was used to explore the purpose of level I fieldwork among occupational therapy students, clinical educators, and faculty respondents at one academic program. Differences in purposes among the three groups of respondents created different fieldwork expectations and outcomes. These differences underlined the importance of communication among students, clinical supervisors, and faculty in planning fieldwork to meet the needs of all three groups. Interpersonal skills, rather than academic skills, emerged as most important to student success in clinical education. Other factors that promote optimal level I fieldwork experience are understanding the purpose, level of commitment, clarity of expectations, timing, structure, and evaluation of the experience. PMID- 8418679 TI - Positioning the nursing home resident: an issue of quality of life. AB - Occupational therapy intervention in the area of seating and positioning may play a vital role in improving the quality of life for nursing home residents. This case report indicates that appropriately positioning a client may increase comfort, decrease agitation, and decrease the administration of mood-altering drugs. Research would help to delineate the effects of appropriate seating systems, both to ensure reimbursement and to ensure that all who might benefit from positioning intervention receive the appropriate services. OBRA 1987 regulations are forcing nursing homes to assess residents for the least restrictive restraints. These assessments offer a golden opportunity for occupational therapists to become involved in determining the most appropriate seating systems and to conduct research on their benefits. PMID- 8418680 TI - Torticollis: a head and neck immobilizer. PMID- 8418681 TI - Academically based education versus continuing education: the best way to go. PMID- 8418682 TI - Maintaining professional education in community settings. PMID- 8418683 TI - Different views are healthy: formulating questions is everything. PMID- 8418684 TI - Pursuing old paths may not be worthwhile. PMID- 8418685 TI - Ketorolac prevents postoperative small intestinal ileus in rats. AB - The effect of ketorolac, a parenterally administered, nonsteroidal anti inflammatory drug, was examined in a rat model of postoperative ileus. Small intestinal transit was measured by calculating the geometric center (GC) of distribution of 51CrO4. Laparotomy significantly delayed transit (GC: 2.2 +/- 0.2 after laparotomy versus 5.6 +/- 0.5 for unoperated controls, p < 0.01). The administration of ketorolac (1 mg/kg) improved the GC to 5.2 +/- 0.2 (p < 0.01), indicating normal intestinal transit after surgery in ketorolac-treated animals. Small intestinal myoelectric activity was recorded in rats with implanted electrodes. Animals treated with saline 2 hours postoperatively did not show return of the migrating myoelectric complex (MMC) in 183 +/- 25 minutes. In contrast, rats receiving ketorolac postoperatively had return of MMC activity in 59 +/- 18 minutes (p < 0.01). Preoperative ketorolac treatment reduced the duration of MMC inhibition after surgery from 197 +/- 55 minutes to 13 +/- 5 minutes (p < 0.05) when compared with saline. In summary, ketorolac hastens the return of MMC activity when given postoperatively. When ketorolac is administered preoperatively, it completely prevents the delay in intestinal transit and the inhibition of myoelectric activity seen in postoperative ileus. We concluded that ketorolac may be of benefit in the prevention and treatment of postoperative ileus. PMID- 8418686 TI - Treatment of acute postoperative ileus with octreotide. AB - Our hypothesis was that postoperative ileus is caused by the release of neurotransmitters in the gut wall that inhibit motility. We reasoned that blocking the release with octreotide would prevent ileus. We placed serosal electrodes on the small intestine and stomachs of 15 dogs and inserted a duodenal cannula. After the animals recovered, electrical activity was recorded, and small bowel transit, colonic transit, and gastric emptying were studied radiographically and scintigraphically. Ileus was induced by celiotomy and intestinal abrasion. Dogs were randomized to receive on postoperative days 0 through 3 either a placebo (n = 5), octreotide, 1.5 micrograms/kg/8 hr subcutaneously (n = 5), or octreotide 0.83 micrograms/kg/hr intravenously (n = 5). Both doses of octreotide resulted in a faster return to preoperative values of small bowel interdigestive myoelectric activity and transit and colonic transit than did the placebo. The larger dose of octreotide, however, slowed gastric emptying. In conclusion, octreotide shortened the duration of postoperative ileus in the small bowel and colon of dogs. PMID- 8418687 TI - How conservatively can postoperative small bowel obstruction be treated? AB - Although postoperative adhesion ileus is the most common cause of small bowel obstruction in adults, its management remains controversial. We retrospectively studied 297 admissions of 227 patients over a period of 14 years to evaluate our conservative approach in managing adhesion ileus. We found that nonoperative therapy of up to 5 days' duration can be used safely for the majority of patients who present with postoperative intestinal obstruction, including those with complete obstruction. In those patients, who responded to conservative treatment, the obstruction resolved within a mean of 22 hours and a maximum of 5 days. A trial of more than 5 days' duration proved ineffective. The conservative approach resulted in a 73% resolution of obstruction with no significant increase in mortality or in the rate of strangulated bowel. PMID- 8418688 TI - Use of the radiolabeled murine monoclonal antibody, 111In-CYT-103, in the management of colon cancer. AB - Monoclonal antibodies directed at tumor-associated antigens may be useful adjuncts for the management of patients with colorectal cancer. The murine monoclonal antibody, B72.3, binds Tag-72, a cell-surface antigen, which is expressed by colorectal carcinoma cells. We investigated the benefit of indium 111-labeled B72.3, 111In-CYT-103, in localizing the presence and extent of disease in patients with suspected or biopsy-proven primary colorectal cancer and in patients with apparently localized recurrent colorectal adenocarcinoma. Twenty patients were enrolled in this study. Each patient received 1 mg of B72.3 labeled with 4 to 5 mCi of 111In. Patients then underwent planar and single-photon emission computed tomographic imaging 2 to 5 days after infusion. Fifteen patients underwent surgery 1 to 14 days after scanning. There were 11 true positives, 1 false positive, 2 true negatives, and 1 false negative. The 111In CYT-103 scan correctly identified the presence or absence of tumor in the 15 patients in whom biopsies were obtained, for an accuracy rate of 87%. Overall, 111In-CYT-103 supplied clinically useful information regarding the extent of disease that was not previously reported by standard techniques in 33% (5 of 15) of patients who underwent surgical exploration. We conclude that 111In-CYT-103 is a promising imaging agent for patients with potentially resectable recurrences and for those patients with a presumed isolated primary tumor requiring preoperative staging. PMID- 8418689 TI - Role of polyamine biosynthesis during gut mucosal adaptation after burn injury. AB - The induction of ornithine decarboxylase (ODC) mRNA and of ODC enzyme activity are important events in gut repair after cutaneous burn injury. ODC catalyzes the rate-limiting step in the biosynthesis of polyamines that are necessary for normal cell growth; alpha-difluoromethylornithine (DFMO) specifically inhibits ODC activity. The purpose of this study was to examine the role of polyamines in the adaptive response of gut mucosa after burn injury. In experiment 1, male Sprague-Dawley rats (250 to 300 g; n = 6/group) were randomized into sham, 60% burn, or 60% burn plus DFMO. In experiment 2, rats with either a 60% burn or 60% burn plus DFMO treatment received spermidine by gavage. We measured ODC activity, polyamine levels, and DNA content at 0, 12, 24, and 48 hours postburn in the mucosa of both the proximal and distal small intestine. Burn injury produced early atrophy (by 12 hours postburn) of the gut mucosa characterized by decreased mucosal weight and DNA content. Increased ODC activity and polyamine content in both the proximal and distal gut mucosa of burned rats preceded restoration of mucosal weight and DNA content that occurred at 48 hours postburn; these responses were prevented by DFMO treatment. Spermidine administration failed to accelerate gut mucosal recovery after burn injury alone, but oral administration of spermidine reversed the inhibitory action of DFMO on gut mucosal repair. These data suggest that the early increases of gut ODC activity and polyamine levels after burn injury are crucial cellular events for the repair of subsequent gut mucosa. PMID- 8418690 TI - Sequential evaluation of islet cell responses to glucose in the transplanted pancreas in humans. AB - We evaluated the hormonal and metabolic responses of denervated pancreas allografts in nine volunteers 3 to 12 months after the transplant (initial) and again 1 year later (follow-up). Eight of the patients received simultaneous pancreas-kidney transplants. The glucose clamp technique was used to create a square wave of hyperglycemia 5.5 mmol/L above the basal glucose level for 2 hours. A biphasic insulin response was evident in each subject, both initially and at follow-up. The initial plasma insulin response was fourfold higher in patients with pancreas-kidney transplants than in normal volunteers. However, the plasma insulin response of the patients with pancreas-kidney transplants at the follow-up study was more similar to that of the normal controls. The plasma glucagon levels were elevated in follow-up clamp studies. Hepatic glucose production and glucose disposal were similar in both studies. At the follow-up examination only, GLUT4, the major insulin-sensitive glucose transporter, was measured in muscle homogenates by immunoblotting. GLUT4 levels in the patients with pancreas-kidney transplants were only 55% as abundant as in normal volunteers. This may be due, in part, to immunosuppressive therapy or to persistent, albeit reduced, levels of hyperinsulinemia even 2 years after transplantation. We concluded that, despite systemic drainage of the pancreas and immunosuppressive therapy, pancreatic insulin secretion, peripheral insulin levels, and muscle insulin responsiveness are restored toward normal levels approximately 2 years after the transplant. PMID- 8418691 TI - Sepsis impairs gut amino acid absorption. AB - Sepsis has been shown to adversely affect the barrier and metabolic functions of the small intestine as well as to reduce mesenteric blood flow and cause histologic damage. However, the effect of sepsis on gut absorptive function has been largely ignored. In this study, intestinal absorption of arginine and an amino acid analogue, aminoisobutyric acid, was studied using in vivo and in vitro techniques in an experimental model of sepsis. In vivo studies showed a significant impairment in the absorption of both amino acids from the intestinal lumen 24 and 72 hours after cecal ligation and puncture. Uptake of these amino acids by everted gut sacs prepared from septic animals was also significantly reduced. This reduction in absorptive capacity of the gut may limit the ability of enteral feeding alone to supply nutritional requirements during sepsis and may also contribute to the associated morbidity and mortality. PMID- 8418692 TI - Results of the surgical treatment of obesity. AB - A prospective, randomized trial comparing vertical banded gastroplasty (VBG) and vertical gastric bypass (GB) for obesity was completed in 106 patients who did not differ in baseline body mass index (BMI = kg/m2) or length of follow-up. The goal of this surgery was to return patients to within 50% of their ideal weight, i.e., a body mass index less than 35 kg/m2, and to accomplish this while maintaining a low risk for malnutrition as well as other morbidity and mortality. Success was defined as a BMI less than 35 kg/m2 because the mortality risk increases rapidly above this degree of obesity. Surgical failures were encountered in 43% of the 54 patients in the VBG group, all of whom had division between the vertical staple lines. The main causes of failure were stenosis and enlargement of the gastroplasty orifice. Surgery failed in 23% of the GB-treated patients, due to perforation of the vertical staple line. An isolated gastric bypass (IGB) not dependent on staples was performed as the remedial operation for the failures of both VBG and GB. IGB was significantly better than VBG or GB, with a success rate of 83% compared with 39% for VBG and 58% for GB. Subsequent experience since completion of this randomized trial in 54 consecutive patients supports IGB for primary, as well as remedial, operations for the morbidly obese (BMI = 40 to 50 kg/m2), as well as for patients who are super obese (BMI greater than 50 kg/m2). PMID- 8418693 TI - Clinical and functional characterization of high gastroesophageal reflux. AB - In 70 consecutive patients with symptoms of gastroesophageal reflux (GER), we studied lower esophageal sphincter (LES) and upper esophageal sphincter (UES) pressures, esophageal peristalsis, and esophageal acid exposure 5 and 20 cm above the LES. Based on the percentage of time the pH was below 4 in the proximal esophagus, the patients were divided into 2 groups: (1) group A, less than 3%, 48 patients; (2) group B, greater than 3%, 22 patients. LES was weaker (13.5 +/- 6.0 mm Hg in group A versus 7.8 +/- 4.6 mm Hg in group B) and shorter (2.2 +/- 0.7 cm in group A versus 1.5 +/- 0.5 cm in group B) in group B patients. Group B patients also had lower amplitude of peristalsis in the proximal (59.2 +/- 17.6 mm Hg in group A versus 42.4 +/- 18 mm Hg in group B) and distal esophagus (89.7 +/- 25.2 mm Hg in group A versus 54.7 +/- 27.9 mm Hg in group B) and lower UES pressures (73.7 +/- 30.7 mm Hg in group A versus 54.7 +/- 29.3 mm Hg in group B). Forty-one percent of group B patients had pulmonary aspiration, whereas only 2% of the patients in group A experienced pulmonary aspiration. These data show that, in a subgroup of patients with symptoms of GER, the upper esophagus is exposed to acid for more than 3% of the time. Patients with high GER differ clinically and pathophysiologically from those in whom reflux is limited to the distal part of the esophagus: those with high GER have a panesophageal motor dysfunction and a high risk of aspiration. PMID- 8418694 TI - Abnormal plasma gut hormones in pathologic duodenogastric reflux and their response to surgery. AB - Fasting and postprandial plasma levels of the gut hormones gastrin, cholecystokinin (CCK), secretin, glucose-dependent insulinotropic polypeptide, motilin, neurotensin, peptide YY (PYY), enteroglucagon, glucagon, insulin, and pancreatic polypeptide were measured in 11 patients with alkaline gastritis associated with excessive duodenogastric reflux not related to previous gastric surgery (primary DGR), 12 primary DGR patients after pancreatico-biliary diversion ("duodenal switch" procedure), and in 10 age-matched healthy controls. Gastric emptying of a semisolid oatmeal was also measured in patients with primary DGR and in patients after bile diversion. Fasting plasma levels of the distal gut hormone neurotensin and the pancreatic islet hormone insulin were significantly greater in patients with primary DGR compared with controls. Neurotensin levels were normal in patients studied after bile diversion. Postprandial plasma levels, incremental integrated and total integrated responses for CCK, secretin, insulin, neurotensin, PYY, and enteroglucagon, were significantly greater in patients with primary DGR compared with controls. The majority of these responses normalized after bile diversion; however, the postprandial response for insulin and enteroglucagon remained elevated. Patients with primary DGR had a rapid early postprandial phase of gastric emptying of solids, which showed a significant correlation with plasma neurotensin levels. Bile diversion produced a significant delay in this lag-phase of gastric emptying. These abnormalities in gut regulatory hormones appear to be adaptive changes to rapid early postprandial gastric emptying, probably related to antropyloric dysmotility, which has been implicated in the pathogenesis of this condition. Measurement of these gastrointestinal hormones may become useful in the diagnosis of primary DGR. PMID- 8418695 TI - Randomized trial of intraoperative radiotherapy in carcinoma of the stomach. AB - A prospectively randomized controlled clinical trial was performed comparing surgical resection and intraoperative radiotherapy (IORT) with conventional therapy in adenocarcinoma of the stomach. Patients in the experimental group underwent gastrectomy, and IORT was administered to their gastric bed (20 Gy using 11 to 15 MeV electrons). Patients in the control group underwent gastrectomy alone for early-stage lesions confined to the stomach (stages I to II) or resection and postoperative external beam radiotherapy to the upper abdomen (50 Gy using 6 to 10 mV photons) for advanced-stage lesions extending beyond the gastric wall (stages III to IV). One hundred patients were screened for the study, of whom 60 were randomized and underwent exploratory surgery. Nineteen patients were excluded intraoperatively because of unresectability or metastatic disease, leaving 41 patients in the study. Seven patients (17%) died of complications. The median survival for patients with tumors of all stages was 25 months for the IORT group and 21 months for the control group (p = 0.99). Locoregional disease failures occurred in 7 of 16 (44%) IORT patients and in 23 of 25 (92%) control patients (p < 0.001). Complication rates were similar between IORT and control patients. Although IORT failed to afford a significant advantage over conventional therapy in overall survival, IORT did significantly improve control of locoregional disease. PMID- 8418696 TI - Continuous duodenal feeding restores gut blood flow and increases gut oxygen utilization during PEEP ventilation for lung injury. AB - Positive end-expiratory pressure (PEEP) improves oxygenation but, at moderate levels, limits portal blood flow (PBF) and may cause relative splanchnic ischemia. Under these conditions, methods of supporting gut physiology may prevent the sequelae of gut ischemic damage. Enteral feeding is known to cause splanchnic hyperemia in uninjured animals. In order to study the effects of continuous enteral feeding on gut hemodynamics in a flow-limited environment, six dogs underwent the insertion of arterial, pulmonary artery, and portal and hepatic vein catheters. Splenectomy and duodenostomy were performed, and the hepatic artery and portal vein were encircled with flow probes. Lung injury (LI) was undertaken with intravenous oleic acid (0.08 mL/kg), followed by incremental additions of PEEP totaling 10 cm H2O to correct shunt. Continuous elemental feeding (1 kcal/mL, 3 mL/kg/hr) was started through the duodenostomy. Cardiac index (CI), PBF, and gut oxygen delivery and consumption (GO2D, GO2C) were measured at baseline (T0), 1 hour after LI and PEEP (T1), and 1 hour after drip feeding was begun (T2). Lung injury and PEEP significantly decreased CI, PBF, and GO2D without changing GO2C. Feeding returned PBF and GO2D to baseline levels without changing CI. GO2C increased significantly compared with baseline levels. Based on prior studies, these changes do not represent recovery of the injured model. Continuous enteral feeding, therefore, redistributed CI to the portal circulation. The improved gut hemodynamics documented in this model may preserve splanchnic integrity and prevent gut-derived complications. PMID- 8418697 TI - Reduction of bacterial translocation with oral fibroblast growth factor and sucralfate. AB - The aim of this study was to evaluate the ability of basic fibroblast growth factor (bFGF) and sucralfate to prevent bacterial translocation after burn injury. Four groups of Balb/c mice (n = 10/group) were treated by gavage with bFGF (10 micrograms/kg/d), sucralfate (15 mg/kg/d), bFGF plus sucralfate, or saline for 4 days prior to receiving a gavage with 1 x 10(10) 14C-radiolabeled Escherichia coli and a 20% full-thickness burn. Four hours after burn, the mesenteric lymph nodes, liver, spleen, and blood were harvested aseptically. For each tissue, the number of viable bacteria and radionuclide counts of the translocated 14C-labeled E. coli were measured, and the percentage of translocated organisms that remained alive was calculated. The results indicated that treatment with either bFGF or sucralfate alone had a partial effect on translocation, whereas the combined treatment with bFGF plus sucralfate significantly decreased the magnitude of translocation in all tested tissues (p < 0.05, ANOVA), which was associated with complete preservation of gut mucosal integrity. None of the treatments affected the ability of the host to kill translocated bacteria when the results were compared with those of the controls. The additive effect of the combined therapy may be due to the high affinity of sucralfate for bFGF, decreasing the degradation of bFGF by gastric acid. PMID- 8418698 TI - Is fellowship training in alimentary tract surgery necessary? PMID- 8418699 TI - Initiation of gastrin expression during the development of the mouse pancreas. AB - Gastrin expression occurs in the pancreas in only two situations: (1) in cases of gastrinoma and (2) in the embryonic/fetal pancreas. The initiation of gastrin expression in the embryonic pancreas may be recapitulated during gastrinoma tumorigenesis. For this reason, we have tried to identify the point of onset of gastrin expression in the developing pancreas. Previously, determining the point of onset for genes in embryonic tissues has been difficult because of low expression levels and small tissue samples. Using the sensitive polymerase chain reaction assay, we were able to determine, with a sensitivity of 10 molecules of mRNA, the earliest expression of gastrin in the developing pancreas. This expression occurred at 30 somites, or at a gestation of 9.5 to 10 days. PMID- 8418700 TI - Shunt surgery versus endoscopic sclerotherapy for variceal hemorrhage: late results of a randomized trial. AB - Between September 1982 and April 1988, 60 cirrhotic patients with prior variceal hemorrhage were randomized to undergo the placement of an elective shunt (distal splenorenal: 26; nonselective: 4) or long-term endoscopic sclerotherapy (n = 30). Eighty-six percent of patients had alcoholic cirrhosis, and 33% were classified as Child's class C. After a mean follow-up of 87 months, 60% of patients undergoing sclerotherapy and 17% of shunt patients experienced rebleeding (p < 0.001). Shunt patients have survived longer than those who had sclerotherapy (6 year survival rates of 53% and 26%, respectively; p < 0.05). In part because of the wide geographic distribution of patients, only 4 of 13 patients in whom sclerotherapy failed (31%) could undergo salvage by shunt surgery. Although hepatic portal perfusion was better maintained after sclerotherapy, there were no major differences between the groups in terms of post-therapy hepatic or psychoneurologic function. In a predominantly alcoholic cirrhotic patient population (half non-urban), the results of elective shunt surgery were superior to those of chronic endoscopic sclerotherapy with respect to the prevention of recurrent variceal hemorrhage and survival. PMID- 8418701 TI - Dexamethasone stimulation of glutaminase expression in mesenteric lymph nodes. AB - Lymphocytes in the mesenteric lymph nodes (MLNs) play a key role in protecting the body from the translocation of bacteria through the bowel during catabolic states. Lymphocytes use glutamine for energy and for DNA synthesis and have high levels of the glutaminase (GA) enzyme that regulates intracellular glutamine metabolism. This study tested the hypothesis that the increase in circulating glucocorticoid hormones that occurs in response to stress states regulates MLN lymphocyte GA expression at the molecular level. Adult male rats (n = 60) received a single dose of dexamethasone (DEX 0.5 mg/kg intraperitoneally) or saline vehicle (CONT). Ileocolic lymph nodes were excised from anesthetized rats via laparotomy at 2, 4, 12, 24, 48, and 72 hours after injection, and GA activity was assayed in MLN homogenates. A second group of rats received repeated doses of DEX (0.5 mg/kg/d for 4 days). GA activity was assayed in MLN homogenates, and total RNA was extracted for quantitation by Northern hybridization using a phosphorus 32-labeled rat GA cDNA probe. GA activity was increased within 2 hours after a single dose of DEX, with a peak after 4 hours, Kinetic analysis at the 4 hour time point showed an increase in the maximum GA activity (maximal transport velocity [Vmax]: 619 +/- 107 nmol/mg protein/hr in DEX versus 380 +/- 53 nmol/mg protein/hr in CONT, p < 0.05), with no change in GA affinity (Michaelis constant [Km]: 2.28 +/- 0.39 mM in DEX versus 2.20 +/- 0.36 mM in CONT, p = NS). Repeated doses of DEX resulted in a twofold increase in GA activity (550 +/- 125 nmol/mg protein/hr versus 1,175 +/- 40, p < 0.01). Simultaneously, GA mRNA levels were increased by 70%. Glucocorticoids stimulate GA activity and gene expression in lymphocytes that reside in MLNs. PMID- 8418702 TI - Pathogenesis of pancreatic sepsis. AB - Although pancreatic sepsis is the most common cause of major morbidity and mortality associated with acute pancreatitis, the pathogenesis of such infections is unknown. Since intraperitoneal foci of inflammation are known to promote bacterial translocation, we hypothesized that acute pancreatitis promotes bacterial translocation that leads to infection of the inflamed pancreas and peripancreatic tissues. Non-lethal acute pancreatitis was induced in rats, and the translocation of live bacteria to the pancreas, mesenteric lymph nodes, liver, and spleen was determined. The presence of orally fed fluorescent beads, sensitive inert markers of translocation, was also determined in the pancreas and mesenteric lymph nodes. Live bacteria were recovered from 33% of the pancreata of rats with acute pancreatitis but from none of the control rats. Beads were visualized in 91% of the pancreata of rats with acute pancreatitis but in none of the pancreata from control rats. Beads were not visualized in the mesenteric lymph nodes of rats with acute pancreatitis, suggesting a transperitoneal route of migration. We conclude that acute pancreatitis promotes bacterial translocation leading to transperitoneal infection of the pancreas. These results support the use of selective decontamination of the gut and peritoneal lavage for the prevention of pancreatic infections in acute pancreatitis. PMID- 8418703 TI - Cost-benefit analysis of the work-up for pancreatic cancer. AB - We reviewed the records of 126 patients with pancreatic cancer to assess the value of diagnostic tests. The most commonly performed studies were computed tomography (CT) (97% of patients), endoscopic retrograde cholangiopancreatography (ERCP) (44%), and fine-needle aspiration (FNA) (41%). Of 34 patients who were found to have a mass in the body or tail of the pancreas on CT, 13 underwent ERCP; the results found by ERCP did not affect the management of the patients, whereas the results of FNA in 12 patients eliminated the need for operation. Of 14 patients with suspected metastases as evidenced by CT, the results of 3 ERCPs had no impact, whereas 5 of 7 patients who had FNAs avoided operation. Of five patients with normal results on CT, three had an ERCP that identified tumor. Of 26 patients with atypical CTs, the results of 12 of 16 ERCPs and 3 of 5 FNAs confirmed cancer. In contrast, in 48 patients with a mass in the head of the pancreas and biliary dilatation, ERCP did not alter the patients' management; only 3 of 14 patients who had FNAs avoided operation. Thus, the results of ERCP rarely altered the management of the patient when the CT showed a mass but was useful when the scan was normal or atypical. FNA was helpful in patients with cancer in the body or tail of the pancreas or with suspected metastases and in confirming the diagnosis when the CT was inconclusive. PMID- 8418704 TI - Comparisons of dynamic infusion and delayed computed tomography, intraoperative ultrasound, and palpation in the diagnosis of liver metastases. AB - The purpose of this study was to determine whether delayed computed tomography (DCT) of the liver would more accurately detect hepatic malignancy when compared with bolus contrast-enhanced dynamic computed tomography (BCDCT). Fifty-one patients who required operation for intra-abdominal malignancy (92% with colorectal cancers) underwent preoperative BCDCT followed by DCT. At operation, palpation and intraoperative ultrasound (IOUS) examination of the liver were performed for localization and biopsy of tumor nodules. The standard for diagnosis was defined for this study as the combined results of IOUS, palpation, and biopsy. The sensitivities of BCDCT and DCT for hepatic metastases were 50% and 54%, respectively, with a corresponding specificity of 72% for each. DCT demonstrated no significant improvement over BCDCT in the detection of individual hepatic lesions. The sensitivity of palpation for the detection of metastases was 82%, equal to that of IOUS. Both palpation and IOUS were significantly superior to BCDCT or DCT in excluding false-positive and false-negative results (p < 0.001). IOUS failed to identify surface lesions less than 1.0 cm in diameter (sensitivity: 40%). Conversely, palpation was limited in the detection of subsurface tumors less than 1.0 cm in diameter (sensitivity: 33%). Combined IOUS and palpation were significantly more accurate in the detection of hepatic metastases than any single modality that was evaluated (p < 0.001). PMID- 8418705 TI - Complications of laparoscopic cholecystectomy: a national survey of 4,292 hospitals and an analysis of 77,604 cases. AB - Complications of laparoscopic cholecystectomy were evaluated by a survey of surgical department chairpersons at 4,292 US hospitals. The 77,604 cases were reported by 1,750 respondents. Laparotomy was required for treatment of a complication in 1.2% of patients. The mean rate of bile duct injury (exclusive of cystic duct) was 0.6% and was significantly lower at institutions that had performed more than 100 cases. Bile duct injuries were recognized postoperatively in half of the cases and most frequently required anastomotic repair. Intraoperative cholangiography was practiced selectively by 52% of the respondents and routinely by 31%. Bowel and vascular injuries, which occurred in 0.14% and 0.25% of cases, respectively, were the most lethal complications. Postoperative bile leak was recognized in 0.3% of patients, most commonly originating from the cystic duct. Eighteen of 33 postoperative deaths resulted from operative injury. These data demonstrate that laparoscopic cholecystectomy is associated with low rates of morbidity and mortality but a significant rate of bile duct injury. PMID- 8418706 TI - Effect of verapamil on hepatic ischemia/reperfusion injury. AB - Alterations in cellular calcium homeostasis are a critical factor in the pathogenesis of hepatic ischemic damage and may mediate oxygen free radical injury during the reperfusion period. We investigated the effect of the calcium channel blocker verapamil on hepatic ischemia/reperfusion injury in normal rats and rats sensitized to oxidative injury by chemical depletion of the endogenous antioxidant glutathione. Forty-five minutes of complete hepatic ischemia followed by reperfusion caused an increase in the serum glutamic pyruvic transaminase (SGPT) level and a decline in the endogenous hepatic glutathione level but no increase in hepatic lipid peroxidation products. Chemical depletion of hepatic glutathione with diethylmaleate did not induce hepatocellular injury but augmented hepatic ischemia/reperfusion-induced SGPT release and promoted lipid peroxidation. Pretreatment with the calcium entry blocker verapamil protected against the ischemia/reperfusion-induced drop in hepatic glutathione but did not reduce SGPT release in normal rats. In rats sensitized to oxidative injury by chemical depletion of endogenous glutathione, the calcium channel blocker verapamil protected against ischemia/reperfusion-induced lipid peroxidation and reduced the release of SGPT. These findings indicate that the rat liver is protected against oxidative injury after short periods of total ischemia by its rich supply of endogenous glutathione. A beneficial effect of verapamil occurs only in rats sensitized to oxidative injury, suggesting that the calcium channel blocker protects against oxygen radical attack. PMID- 8418707 TI - Effect site equilibration time is a determinant of induction dose requirement. PMID- 8418708 TI - Determinants of thiopental induction dose requirements. AB - Dose requirements for thiopental anesthetic induction have significant age- and gender-related variability. We studied the association of the patient characteristics age, gender, weight, lean body mass, and cardiac output with thiopental requirements. Doses of thiopental, infused at 150 mg/min, required to reach both a clinical end-point and an electroencephalographic (EEG) end-point were determined in 30 males and 30 females, aged 18-83 yr. Univariate least squares linear regression analysis revealed outliers in the relationships of age, weight, lean body mass, and cardiac output to thiopental dose at clinical and EEG endpoints. Differential weighting of data points minimized the effect of outliers in the construction of a robust multiple linear regression model of the relationship between several selected independent variables and the dependent variables thiopental dose at clinical and EEG endpoints. The multiple linear regression model for thiopental dose at the clinical end-point selecting the regressor variables age, weight, and gender (R2 = 0.76) was similar to that for age, lean body mass, and gender (R2 = 0.75). Thiopental dose at the EEG endpoint was better described by models selecting the variables age, weight, and cardiac output (R2 = 0.88) or age, lean body mass, and cardiac output (R2 = 0.87). Although cardiac output varied with age, age always remained a selected variable. Because weight and lean body mass differed with gender, their selection as variables in the model eliminated gender as a selected variable or minimized its importance. PMID- 8418709 TI - Subcutaneous recombinant human erythropoietin and autologous blood donation before coronary artery bypass surgery. AB - Conventional therapies with recombinant human erythropoietin (rHuEPO) to sustain preoperative autologous blood collection entail high doses of the drug at short intervals. To evaluate the efficacy of a single weekly dose of rHuEPO for autologous blood collection, we randomly assigned 24 male patients scheduled for coronary artery bypass surgery to receive 400 IU/kg rHuEPO subcutaneously once a week or iron only. Patients were examined weekly and a total of up to 4 units of autologous blood were obtained if the hemoglobin level exceeded 12 g/dL. Patients receiving rHuEPO had consistently higher hemoglobin values than those receiving iron only (P < 0.001). Consequently, more autologous red cells were obtained from this group (776 +/- 49 mL vs 682 +/- 91 mL; P < 0.05). One patient receiving rHuEPO and eight in the control group required homologous blood at surgery (P < 0.01). These results suggest that 400 IU/kg rHuEPO administered subcutaneously once a week efficiently stimulates erythropoiesis and compensates the hemoglobin decrease after autologous blood donation. PMID- 8418710 TI - Quantitative analysis of pulmonary clearance of exogenous dopamine after cardiopulmonary bypass in humans. AB - The contribution of the lung to the clearance of exogenous dopamine after cardiopulmonary bypass (CPB) was analyzed quantitatively in humans and compared with the contribution of the lung before CPB. The pulmonary and arterial plasma concentration of dopamine and the pulmonary plasma flow were measured simultaneously during infusion of dopamine. Contribution of the pulmonary circulation was defined as the ratio between clearance through the pulmonary circulation and the total plasma clearance of dopamine. The calculated contribution values after CPB were 12.0, 10.7, 11.4, 16.2, and 16.7% at the doses of 3.0, 4.0, 5.0, 6.0, and 7.0 micrograms.kg-1.min-1, respectively. Those values before CPB were 15.6% and 17.4% at the doses of 1.0 and 2.0 micrograms.kg-1.min 1, respectively. The comparison of the values before and after CPB did not achieve statistical significance. Furthermore, there were no significant correlations between the pulmonary clearance after CPB and mean pulmonary arterial pressure, pulmonary vascular resistance, or CPB time. The results suggest that the pulmonary clearance mechanism for dopamine after CPB is maintained as effectively as that before CPB and is not influenced by pulmonary hypertension or CPB time. PMID- 8418711 TI - Does timing of hemodilution influence the stress response and overall outcome? AB - This study was designed to assess the stress response of acute hemodilution (AH) in patients subjected to radical cystectomy. Forty adult male patients were randomly allocated into a control group (n = 10) where homologous blood transfusion was used, a preinduction AH group (n = 20) where AH was performed before lumbar epidural block and induction of anesthesia, and into a postinduction group (n = 10) where AH was performed after induction of anesthesia. Monitored variables included hemodynamic, hematological and coagulation factors, liver function tests, and serum hormones. AH performed in awake or in anesthetized patients did not result in significant hemodynamic disruption, or result in detectable end-organ or stress-hormone changes when compared to control patient outcomes after radical cystectomy. Hemodilution can be performed by protocol for patients who are undergoing this procedure without major adverse effects. PMID- 8418712 TI - Median nerve blockade during diagnostic intravenous regional anesthesia as measured by somatosensory evoked potentials. AB - Intravenous regional anesthesia (IVRA) may be used as a diagnostic method in patients suffering from chronic pain in the forearm or hand to differentiate the origin of pain within the anesthetic area from that above. For this purpose it needs to be proven that all nerve fibers are blocked and that conduction blockade induced by IVRA takes place within the nerve trunks. Therefore the transmission of impulses in a nerve trunk to the central nervous system has been studied. Diagnostic intravenous regional anesthesia (5 mg/kg mepivacaine 1%) of the arm was performed in eight patients for 30 min. Short-latency somatosensory evoked potentials (SSEPs) induced by median nerve stimulation at the wrist were recorded from the scalp at 5-min intervals before, during, and after IVRA. In five patients, sensory nerve action potentials (SNAPs) of the median nerve, at the elbow and axilla, and SSEPs at spinous processus C-7 were recorded simultaneously. During IVRA the function of the nerve fibers which are assumed to mediate pain was tested by the patient's sensation following median nerve stimulation at an intensity which evoked pain before IVRA. During IVRA, peak latencies of the scalp recorded SSEPs (N20) increased progressively and interpeak amplitudes (N20/P25) decreased. After 20 min, SSEPs could no longer be recorded, and median nerve stimulation no longer evoked any sensation at all. After deflation of the cuff, both peak latency (N20) and interpeak amplitude (N20/P25) of SSEPs recovered. The changes in latency and amplitude of SSEPs from the scalp as well as SNAPs and SSEPs from the neck were similar. Because SSEPs, SNAPs, and the pain sensation following median nerve stimulation disappeared during IVRA, it may be concluded that the thick and thin myelinated nerve fibers of the median nerve have been blocked. For diagnostic use, IVRA is superior to peripheral nerve blockade, which has been shown previously to not abolish SSEPs. PMID- 8418713 TI - Acute toxicology of an enkephalinase inhibitor (SCH 32615) given intrathecally in the ewe. AB - Intrathecal application of the enkephalinase inhibitor, SCH 32615, yields antinociception in animal paradigms. Our purpose was to identify possible acute behavioral effects, neurotoxicity, or systemic toxicity of intrathecal SCH 32615 administration during 9 days in the ewe. Seventeen ewes were implanted with lumbar silicone intrathecal catheters and subcutaneous access ports for repeated injection. Baseline and serial daily behavioral assessments were made during 9 days of 2-mL intrathecal injection twice daily of either normal saline (SAL group) or a 20 mg/mL isotonic sterile solution of SCH 32615 (SCH group). Data were analyzed by treatment group (SCH versus SAL) by taking the group means of individual ewe cumulative scores during 9 days. At 15-18 h after the last injection, the ewes were euthanized and the spinal cords and leptomeninges were grossly examined and prepared for histological assessment. Histological evaluation of the lumbar (at catheter entrance site and catheter tip), thoracic, and cervical sections of all animals was performed by two neuropathologists. Several mild, reversible, and apparently nonprogressive behaviors (Stepping/Placing and Hindlimb Stretching/Splaying) were observed almost exclusively in SCH-treated ewes. These behaviors were interpreted as mild temporary irritative effects, without significant neuropathological sequelae. Pathological findings primarily consisted of mild, focal dural thickening and white matter compression. These changes were distributed equally between drug treated and control groups and were attributable to catheter implantation and local compressive effects. There were no pathological bases identified in this study to preclude the clinical study of SCH 32615 within the dose range studied. PMID- 8418714 TI - On the mechanisms of potentiation of local anesthetics by bicarbonate buffer: drug structure-activity studies on isolated peripheral nerve. AB - Impulse inhibition by local anesthetics (LAs) is potentiated by extracellular solutions containing HCO3-. CO2 (BC), relative to the inhibition in BC-free solutions at the same pH. We studied the mechanistic basis of this potentiation by assaying compound action potential amplitudes in desheathed frog sciatic nerves with the sucrose-gap method. We compared the potencies of 12 different impulse-blocking agents in Ringer's buffered with BC (BC-R) and in Ringer's containing only atmospheric CO2 and buffered by a zwitterionic compound (3-(N morpholino)propanesulfonic acid-Ringer's). The relative inhibition produced by an agent in BC divided by the inhibition produced in 3-(N-morpholino)propanesulfonic acid, was defined as the potentiation factor (PF). The organic guanidinium blockers of sodium channels, tetrodotoxin and saxitoxin, which act at a different site from that for LAs, were, by our definition, nominally potentiated (PF = 1.33 +/- 0.04, mean +/- SEM, n = 4, and 1.24 +/- 0.07, n = 10, respectively), implying that BC induces a decrease in the safety margin for impulse conduction, a decrease that cannot itself alone account for the much larger potentiation (PF = 5-8) by BC observed with certain LAs. Only nominal potentiations occurred with charged LAs (PF = 1.15), showing that little direct potentiation of the cationic LA species per se occurs. Inhibition by the permanently neutral LA benzocaine had a significantly larger than nominal potentiation (PF = 1.8) showing that BC can potentiate neutral LAs. Among the tertiary amine LAs, potentiation of ester linked drugs (procaine, RAG505; PF = 3.9, 5.4, respectively), exceeded that of their amide-linked homologues (procainamide, lidocaine; PF = 1.3, 2.8, respectively) which have higher pKa values. This result is consistent with an ion trapping mechanism whereby CO2 acidifies the axoplasm and thereby increases the concentration of protonated LA inside the nerve fibers. However, slight differences in the molecular structure of 3 degrees-amine LAs with similar pKa values resulted in significantly different potentiations (e.g., procaine, PF = 3.9; 2-chloroprocaine, PF = 8.7), suggesting that the HCO3- or CO2 molecules interact specifically with the LA molecule or with LA binding sites in the nerve membrane. Spectrophotometric measurements of the free [Ca2+] in Ringer's showed it to be similar (+/- 0.03 mM) for both buffers, obviating changes in extracellular Ca2+ as a mechanism of BC potentiation. The resting potential of the nerve was slightly more negative (approximately -4 mV) in BC-R, so membrane depolarization cannot explain the potentiation.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418715 TI - Propofol anesthesia reduces emesis and airway obstruction in pediatric outpatients. AB - This study was an authors comparison of the effects of and recovery from anesthesia in healthy, premedicated pediatric outpatients who received either inhaled anesthetics (group 1) or propofol (group 2). Group 1 (n = 68) averaged 3.8 +/- 0.2 yr and weighed 17.7 +/- 0.8 kg, whereas group 2 (n = 75) averaged 3.3 +/- 0.2 yr and weighed 16.3 +/- 0.6 kg. The incidence of vomiting in the Postanesthetic Care Unit (PACU) and from discharge to the first postoperative morning was lower in the group receiving propofol (0% and 18%) than in the group receiving volatile agents (7% and 34%, P < 0.05). The incidence of airway obstruction during induction of anesthesia was higher (34% vs 10%, P < 0.01) in children receiving inhaled agent. Withdrawal of the extremity with propofol injection occurred in 14 (19%) patients. Arterial blood pressure was higher at loss of consciousness, laryngoscopy, and tracheal intubation in group 2 (P < 0.01). The length of time from the end of surgery to extubation of the trachea, recovery scores, and length of time spent in the PACU and the Day Surgery Unit were the same in the two groups. Pain scores obtained in the PACU were not different. The data indicate that propofol can be used safely to induce and maintain anesthesia in healthy pediatric outpatients. This coupled with the low incidence of vomiting and airway obstruction in the propofol group suggests distinct and compelling reasons to consider using the drug in this patient population. PMID- 8418716 TI - Pulmonary capillary pressure measurement during global hypoxia in sheep. AB - Analysis of the pressure decay following pulmonary artery occlusion can be used to determine pulmonary capillary pressure and to calculate the magnitudes of the arterial and venous components of pulmonary vascular resistance. The separation of pulmonary vascular resistance into components has been termed "the longitudinal distribution of pulmonary vascular resistance" to emphasize the fact that different pressures occur at a number of sites in the pulmonary circulation. The longitudinal distribution of pulmonary vascular resistance is closely related to pulmonary capillary pressure. Several methods of data analysis have been proposed to determine pulmonary capillary pressure from the pressure decay following pulmonary artery occlusion. In this study, three methods of data analysis were applied to the model of hypoxic pulmonary vasoconstriction to evaluate the validity of the methodology in a well known model. Pulmonary artery occlusion pressure decay curves were obtained from eight halothane-anesthetized sheep during control conditions (FIO2 = 0.99) and during hypoxic ventilation (FIO2 = 0.14). Analysis of the pulmonary artery occlusion pressure decay curves indicated the following results: 1) Hypoxia increased mean pulmonary artery pressure by 105% and increased pulmonary vascular resistance by 149%; 2) the increase in the calculated arterial component of pulmonary vascular resistance accounted for 88% of the increase in pulmonary vascular resistance with hypoxia; and 3) hypoxia produced only a 1.0 mm Hg increase in pulmonary capillary pressure. These results are consistent with other evidence showing that hypoxia primarily produces precapillary pulmonary vasoconstriction and has little effect on pulmonary capillary pressure. Pulmonary artery occlusion pressure decay curve analysis appears to be a valid technique for the measurement of pulmonary capillary pressure during hypoxia in intact anesthetized animals. PMID- 8418717 TI - Intravenous ranitidine reduces the risk of acid aspiration of gastric contents at emergency cesarean section. AB - This study documented gastric pH and volume and the number of patients at risk of acid aspiration of gastric contents in a group of mothers undergoing emergency cesarean section under general anesthesia. Patients were randomized in a double blind fashion to receive ranitidine, 50 mg intravenously, or placebo at the time of decision to proceed to cesarean section. In addition, all patients received 30 mL of 0.3 M sodium citrate on entry into the operating room. Aspiration of gastric contents was undertaken immediately after endotracheal intubation (PI) and before tracheal extubation. Patients with both pH < 3.5 and volume > 25 mL were deemed to be at risk of acid aspiration should regurgitation occur. Postintubation, 12 patients (4%) were at risk in the citrate-alone group and 7 patients (2.3%) were at risk in the ranitidine/citrate group (not significant). Preextubation, 17 patients (5.6%) were at risk in the citrate-alone group and 1 patient (0.3%) was at risk in the ranitidine/citrate group (P < 0.05). PI pH in patients receiving ranitidine/citrate (mean 5.2, SD 0.8) was significantly higher than in patients receiving citrate alone (mean 4.9, SD 1.1). None of the patients who received ranitidine more than 30 min before the PI sample were at risk compared to 6 (3.2%) in the citrate alone group (P = 0.05). We conclude that 50 mg of intravenous ranitidine given at the time of decision to proceed to cesarean section reduces the risk of acid aspiration provided that at least 30 min have elapsed from injection to induction of anesthesia. PMID- 8418718 TI - Electrocardiographic changes during cesarean section: a cause for concern? AB - A Holter monitor was used to record ST segment changes during cesarean section in 170 consecutive healthy parturients starting 2 h before and ending 3 h after surgery. Lumbar epidural anesthesia (LEA, n = 120) or subarachnoid anesthesia (SA, n = 50) was used. Transthoracic 2-D echocardiograms were obtained in 30 patients from the LEA group. ST depression or elevation occurred 160 times in 44 patients from both groups. Ninety-eight percent of these changes occurred between induction of anesthesia and the end of surgery, with 78% of the episodes registering -1 mV. In the LEA group, the number of episodes tended to increase after delivery, but in the SA group, the frequency remained constant. ST segment depression was recorded in 38% and 14% of patients in the LEA and SA groups, respectively (P < 0.05, x2 analysis). No wall motion abnormality was noted in the echocardiogram during ST segment depression. Neither the 12-lead electrocardiogram nor plasma myocardial specific creatine kinase suggested myocardial damage. The operative events, alone or in combination, including hypertension, tachycardia, hypotension, bradycardia, air embolism (precordial Doppler) were neither specific nor sensitive as predictors of ST segment change (stepwise logistic regression). Tachycardia was associated with ST segment changes in 10% of time epochs (5 min) (P = 0.05, x2 analysis). Thus, ST segment changes during cesarean section are not caused by myocardial ischemia and are not of any clinical consequence. PMID- 8418719 TI - Mild hypothermia does not impair postanesthetic recovery in infants and children. AB - We tested the hypothesis that mild perioperative hypothermia (e.g., central temperature 34-36 degrees C) does not impair postanesthetic recovery in infants and children. The postoperative course was evaluated in 84 ASA Physical Status 1 or 2 patients, weighing 5-50 kg, who were recovering from peripheral surgery lasting less than 3 h. Postoperative monitoring initiated in the Post Anesthesia Care Unit (PACU) included: 1) rectal (central) temperature; 2) oxyhemoglobin saturation (Spo2) while breathing 21% oxygen; 3) apnea (using impedance pneumography); and 4) duration of recovery determined by an anesthesiologist not involved with the study and unaware of the patients' central temperatures. Data were stratified according to patients' weights and central temperatures on admission to PACU. Recovery was rapid in all patients, with no statistically significant differences in duration of recovery among the groups. Only three patients had Spo2 values less than 90%, and in each case desaturation was observed during only one 15-min epoch and resolved without specific treatment. The lowest observed Spo2 was 82%. These three patients weighed between 10 and 20 kg and had initial temperatures exceeding 36 degrees C. No patient had an apneic episode exceeding 15 s. We conclude that mild hypothermia per se neither impairs respiratory function, nor prolongs postanesthetic recovery in generally infants and children undergoing peripheral surgery. PMID- 8418720 TI - Postoperative voiding interval and duration of analgesia following peripheral or caudal nerve blocks in children. AB - We studied the time to postoperative micturition and the duration of analgesia in 82 children aged 6 mo to 10 yr undergoing herniorrhaphy or orchiopexy under general anesthesia with N2O and halothane. All received D5 lactate Ringer's solution equivalent to 6 h maintenance intraoperatively, and oral fluids postoperatively ad libitum. At the end of surgery, patients were randomly assigned to receive one of three regional anesthetic injections using 0.25% bupivacaine: caudal, 0.75 mL/kg (group I); caudal with 1:200,000 epinephrine, 0.75 mL/kg (group II); or ilioinguinaliliohypogastric nerve block with epinephrine through the wound by the surgeon (group III). Postoperatively, blinded observers scored pain at 30 min, hourly until discharge, and by telephone at 24-36 h. In the 74 patients with successful blocks (mean age 2.5 +/- 2.4 yr), the times to micturition (group I, 202 +/- 130 min; group II, 262 +/- 164 min; group III, 196 +/- 101 min) did not differ significantly among groups. Seven patients who took more than 8 h to void required no intervention. There was no difference in the numbers without pain for > or = 4 h (74%, 64%, and 69% of groups I, II, and III), or those requiring analgesics by 24 h (66% overall). The time to postoperative voiding in children is variable and not prolonged by caudal analgesia; caudal bupivacaine with or without epinephrine and ilioinguinaliliohypogastric nerve block are equally effective for postoperative analgesia. PMID- 8418721 TI - Intravenous isoproterenol as a marker for epidural test-dosing in children. AB - The purpose of this study was to determine if isoproterenol would be an effective marker of intravascular injection in anesthetized children. Forty-four ASA 1 children, aged 2 mo to 10 yr, were randomly assigned to two groups. Children in group 1 (n = 21) received 0.05 microgram/kg isoproterenol, and children in group 2 (n = 23) received 0.075 microgram/kg isoproterenol. A blinded observer continuously recorded heart rate and arterial blood pressure. Measurements were recorded before the surgical incision at steady-state halothane concentration of 1.2 minimum alveolar concentration adjusted for age. Isoproterenol produced a graded increase in heart rate with mean maximum increases of 16.5 +/- 8.7 beats/min in group 1 and 21.5 +/- 9.2 beats/min in group 2. No episodes of hypotension and arrhythmia were noted. Isoproterenol, 0.075 microgram/kg, is more sensitive but still is an imperfect marker of an intravascular injection. It produces a heart rate increase in 96% of children anesthetized with halothane and nitrous oxide in 50% oxygen. The application of isoproterenol as an epidural test dose appears promising, but cannot be recommended until its full reliability and neurotoxicity are evaluated. PMID- 8418722 TI - Distinctive effects of three intravenous anesthetics on the inward rectifier (IK1) and the delayed rectifier (IK) potassium currents in myocardium: implications for the mechanism of action. AB - The mechanism(s) of action of anesthetics on cell membrane ionic currents are not known. To investigate this further the effects of clinically relevant concentrations of ketamine, methohexital, and propofol on the delayed rectifier (IK) and the inward rectifier (IK1) currents of single dispersed guinea pig ventricular myocytes were studied. These voltage-gated currents are major components of cardiac cell electrophysiologic function regulating resting potential and repolarization. Each of the three anesthetics had a distinct spectrum of activity. Ketamine (10(-4) M) decreased IK1 (P < 0.05) but had no effect on IK. Methohexital (10(-4) M) had no significant effect on either current. Propofol (2.8 x 10(-5) M) resulted in significant depression of IK (P < 0.001) but had no effect on IK1. These results suggest that these intravenous anesthetics may have more specific effects on sarcolemma than volatile anesthetics, whose effects may be more generalized membrane effects. PMID- 8418723 TI - Prolonged spontaneous movement following emergence from propofol/nitrous oxide anesthesia. PMID- 8418724 TI - Prolonged neuromuscular block after mivacurium. PMID- 8418725 TI - Fetal tracheal intubation with intact uteroplacental circulation. PMID- 8418726 TI - Hypoxia following perioperative administration of vancomycin. PMID- 8418727 TI - Accidental intraspinal overdose revisited. PMID- 8418728 TI - Accidental overdosage of systemic morphine during intended refill of intrathecal infusion device. PMID- 8418729 TI - Detection of intravascularly located epidural catheter. PMID- 8418730 TI - Concealed illegal drugs: a potential cause of airway obstruction. PMID- 8418732 TI - Problems with a spinal catheter. PMID- 8418731 TI - Hypocalcemia following brachial plexus block. PMID- 8418733 TI - Pacemaker and ESWL. PMID- 8418734 TI - What does "resuscitate" mean in a do-not-resuscitate (DNR) order? PMID- 8418735 TI - A use of telescope of rigid type bronchoscope through endotracheal tube in infants. PMID- 8418736 TI - Propofol produces endothelium-independent vasodilation and may act as a Ca2+ channel blocker. AB - The mechanism of vasodilation induced by propofol was investigated using isolated rat thoracic aortic rings. Aortic rings were precontracted with potassium chloride (KCl) (40 mM) or phenylephrine (PE) (3 x 10(-8) to 3 x 10(-7) M) in the presence and absence of intact endothelium. Propofol produced similar concentration-dependent relaxation in aortic rings with and without endothelium regardless of whether they were precontracted with KCl or PE. The relaxation response to propofol was significantly greater in KCl-contracted aortic rings than in PE-contracted aortic rings. The propofol concentration producing 50% relaxation from the contracted state (RC50) was lower in aortic rings contracted with KCl than with PE, both with (5 +/- 0.6 x 10(-5) M vs 8.3 +/- 5.7 x 10(-4) M, P < 0.001) and without intact endothelium (3.9 +/- 0.5 x 10(-5) M vs 7.2 +/- 3.8 x 10(-4) M, P < 0.001). Propofol inhibited the Ca(2+)-induced contractions of aortic rings exposed to Ca(2+)-free media and depolarized with KCl (40 mM, 100 mM) in a dose-dependent manner. These effects are similar to those produced by verapamil. Propofol (5 x 10(-5) M) had minimal effect on the intracellular Ca2+ release elicited by PE (10(-5) M). We conclude that vasodilation produced by propofol is not endothelium-dependent but is likely due to blockade of voltage gated influx of extracellular Ca2+. PMID- 8418737 TI - Effects of propofol on the function of normal, collateral-dependent, and ischemic myocardium. AB - To examine the effects of propofol on the function of normal, collateral dependent, and acutely ischemic myocardium, nine mongrel dogs were chronically instrumented with hydraulic occluders and ameroid constrictors were inserted around the left coronary artery, pressure transducers in the left ventricle, and heparin-filled catheters in the descending aorta and the left atrium. Regional function of normal, collateral-dependent, and acutely ischemic myocardium was assessed by sonomicrometry. Propofol (5 mg/kg intravenously) reduced function in normal myocardium (-15% +/- 5%, 1 min and -14% +/- 5%, 3 min after injection) and in collateral-dependent myocardium (-14% +/- 5% and -13% +/- 5%) to similar degrees, whereas ischemic myocardial function deteriorated significantly more ( 25% +/- 10% and -23% +/- 10%, P < 0.01). Although left ventricular end-diastolic pressure remained unchanged and left ventricular contractility was reduced (-16% +/- 4%, 1 min and -15% +/- 3%, 3 min after propofol, P < 0.01), significant increases in heart rate (35% +/- 7% and 26% +/- 7%, P < 0.01) and decreases in coronary perfusion pressure (-14% +/- 5%, P < 0.05 and -19% +/- 6%, P < 0.01) occurred, likely affecting the function of ischemic myocardium. Thus, whereas collateral-dependent myocardium tolerated these adverse hemodynamic effects, ischemic myocardium responded with impairment of regional function that was significantly more pronounced than the impairment which occurred in normal or collateral-dependent areas after a 5 mg/kg intravenous bolus of propofol. PMID- 8418738 TI - Effect of the head-down tilt position during lower abdominal surgery on endocrine and renal function response. AB - A head-down tilt position in awake subjects induces natriuresis, accompanied by reduced plasma levels of catecholamines, renin, angiotensin II, aldosterone, and antidiuretic hormone. We hypothesized that the head-down tilt position would counteract the surgical stress response which induces sodium and water retention and increases plasma levels of these hormones. We studied endocrine and renal function responses in patients during lower abdominal surgery, performed under sevoflurane anesthesia, at a 6 degree head-down tilt position (n = 10) versus a horizontal position (n = 10). The mean arterial pressure was maintained constant by adjusting the inspired concentration of sevoflurane. Heart rate, and the dose of sevoflurane, decreased significantly in the head-down tilt position. Increases in plasma norepinephrine and epinephrine levels during surgery were significantly less in the head-down tilt position; in contrast, plasma aldosterone and cortisol levels were increased significantly in this position compared to the horizontal position. Both fractional and total urinary excretion of sodium increased significantly in the head-down tilt position compared to the horizontal position. Plasma renin activity, and antidiuretic hormone and atrial natriuretic peptide levels, as well as urine volume, creatinine clearance, and water clearance showed no positional variation. We conclude that the 6 degree head-down tilt position reduces increased sympathetic activity and lessens renal tubular sodium reabsorption during lower abdominal surgery under sevoflurane anesthesia. Other stress hormones, however, increase more in the head-down tilt position than in the horizontal position. PMID- 8418739 TI - Ketamine infusion for postoperative analgesia in asthmatics: a comparison with intermittent meperidine. AB - Narcotics commonly used for postoperative analgesia may release histamine and cause bronchospasm in asthmatics. Ketamine, on the other hand, provides analgesia and has the additional advantage of preventing and relieving bronchospasm. We therefore delivered subanesthetic doses of ketamine in combination with midazolam (5.88-6.42 micrograms.kg-1.min-1 and 1.17-1.28 micrograms.kg-1.min-1, respectively), via an infusion for postoperative analgesia after elective abdominal hysterectomy in patients with asthma. Data were compared with those from a similar group of patients receiving conventional intramuscular meperidine. A significant degree and earlier onset of analgesia (P < 0.05) was achieved in the ketamine group. For other variables no significant difference was observed between the groups (P > 0.05). Ketamine-midazolam infusion can thus provide a safe alternative to the usual parenteral narcotic therapy in asthmatics, in terms of analgesia and patient acceptability. PMID- 8418740 TI - Convective warming therapy does not increase the risk of wound contamination in the operating room. AB - Although convective warming therapy is effective in preventing hypothermia in anesthetized patients, little is known concerning the potential risks of its use. Hence, this balanced cross-over study was designed to determine if the use of convective warming therapy increased the risk of wound contamination. For 4 h, eight healthy male volunteers (aged 20-25 yr) lay supine on an operating room table with their lower bodies and legs covered with a warming cover and sterile surgical drape. The convective warming therapy was administered for 2 h. The other 2 h served as the control. In each session, culture plates were placed directly on the subject's abdomen through an opening in the drape. Tympanic membrane and leg skin temperatures were significantly higher with active warming. No significant differences in the number of bacterial colonies were observed between the two study periods. It was concluded that convective warming therapy, when appropriately applied, does not increase the risk for airborne bacterial wound contamination in the operating room. PMID- 8418741 TI - Halothane reduces release of adenosine, inosine, and lactate with ischemia and reperfusion in isolated hearts. AB - We investigated the protective effects of halothane on cardiac function of isolated hearts during global hypoperfusion and reperfusion by examining halothane's effects on altering coronary flow, myocardial oxygen utilization (MVO2), and release of adenosine (ADE), inosine (INO), and lactate (LAC). Isolated perfused guinea pig hearts were divided into three groups of perfusion at 25% (14 mm Hg), 10% (5.5 mm Hg), and 0% (no perfusion) from control perfusion pressure (PP, 55 mm Hg). Each of these PP groups was subdivided into three subgroups and perfused without halothane (control), with 0.23 +/- 0.01 mM (0.74%) halothane, or with 0.51 +/- 0.01 mM (1.65%) halothane. Halothane was present 10 min before reducing PP, during reduced PP (30 min), and for 10 min after reducing PP. Hypoperfusion was followed by 40 min of reperfusion at the control (100%, 55 mm Hg) PP. An additional group of control hearts was followed for the same period without reducing PP or perfusing with halothane. Exposure to 0.74% and 1.65% halothane, before reducing PP, decreased MVO2 and percent oxygen extraction (% O2E), but produced no significant change in coronary flow or release of ADE, ISO, or LAC. During early hypoperfusion (10 min) at 25% PP, 1.65% halothane significantly reduced release of ADE, INO, and LAC. During late hypoperfusion (40 min) the differences in LAC release diminished, but release of ADE and INO remained lower in the 1.65% halothane group. With early reperfusion there was a large increase in release of these metabolites, that was dependent on the decrease in perfusion pressure.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418742 TI - Halothane myocardial depression: interactions with the adenylate cyclase system. AB - The involvement of the adenylate cyclase system in myocardial depression by halothane was investigated in an isolated, electrically stimulated, rat left atrial preparation. The twitch tension dose-response curve to the muscarinic agent, carbachol, was unaltered by 0.27 mM (0.8%) halothane. Pertussis toxin irreversibly blocks the inhibition of adenylate cyclase by muscarinic agonists. Atria from animals pretreated with pertussis toxin were insensitive to the negative inotropic effect of carbachol, but the depression in twitch tension by halothane was unaltered. Halothane shifted the dose-response curve to isoproterenol downward and to the right. However, the depression in twitch tension by 0.27 mM halothane was similar in preparations incubated with the nonhydrolyzable cAMP analogue, dibutyryl-cAMP, compared to control atria stimulated with isoproterenol. We conclude that halothane attenuates the response to beta-adrenergic stimulation in myocardial tissue, and alterations in adenylate cyclase activity do not contribute significantly to this process. PMID- 8418743 TI - Recovery of function and adenosine triphosphate metabolism following myocardial ischemia induced in the presence of volatile anesthetics. AB - Using a normothermic isolated working rabbit heart model, experiments were performed to determine whether exposure to halothane or isoflurane prior to ischemia improved postischemic recovery of myocardial function and the preservation of myocardial high energy phosphates. After 30 min of Langendorff perfusion, hearts were perfused for 30 min in the working mode. Three groups were studied: 1) the blank undergoing no pretreatment during an additional 15 min of working mode; 2) hearts exposed to 1.5% halothane; and 3) hearts exposed to 2.3% isoflurane during the additional 15 min of working mode. Subsequently, all hearts underwent 15 min of global normothermic ischemia, followed by 5 min of Langendorff reperfusion and 15 min of working heart mode using a perfusate devoid of volatile anesthetic. Adenosine triphosphate (ATP) and catabolites were determined after 15 min exposure to volatile anesthetics or blank, after 15 min global ischemia and at the end of the recovery phase. Myocardial function was determined after 30 min of working mode, after exposure to volatile anesthetics, and at the end of the recovery phase. In nonischemic hearts, 15-min treatment with 1.5% halothane or 2.3% isoflurane resulted in a significant decrease in positive LVdP/dt, from 1858 +/- 286 to 1316 +/- 180 mm Hg.s-1 and from 1888 +/- 304 to 1541 +/- 226 mm Hg.s-1, respectively. Coronary flow was increased significantly after isoflurane but not after halothane.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418744 TI - Postoperative ulnar nerve palsy--is it a preventable complication? PMID- 8418745 TI - Supramaximal second gas effect: more rapid rise of alveolar halothane concentration during ipsilateral lung N2O administration compared to bilateral administration. AB - To elucidate the mechanism of the second gas effect, we enhanced halothane uptake by a method other than by increasing the inspiratory concentration of N2O. We determined the effect of N2O elimination via the right lung, which is not receiving N2O (halothane and oxygen), on the halothane uptake in the left lung with N2O added to an inspiratory gas mixture during a differential ventilation using a double-lumen tube. Under the setting, some N2O which was absorbed in the left lung, and eventually eliminated via the right lung, decreased end-tidal (ET) N2O and thereby increased the inspired to end-tidal gradient for N2O in the left lung which was receiving N2O. The situation thus created was equivalent to administering N2O in a higher concentration than the initial concentration. This process enhanced the halothane uptake in the left lung. The study consisted of 15 patients assigned to three groups with five patients in each group. Control groups received a standard, single-lumen endotracheal tube using a gas mixture of O2 + halothane with and without N2O. The experimental group received a double lumen tube for differential lung ventilation. A N2O + O2 + halothane mixture was administered to the left lung, and simultaneously O2 + halothane was administered to the right lung. On-line gas measurement was performed using Raman spectrometers. The second gas effect was observed between the control groups. N2O was detected in the exhaled gas from the right lung after 3 min of inhalation into the left lung.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418746 TI - The inhibitory effects of positive end-expiratory pressure (PEEP) on phrenic motoneuronal activity in cats. AB - We examined the effects of positive end-expiratory pressure (PEEP) on phrenic single motoneuronal activity (SMA) in anesthetized, paralyzed, and mechanically ventilated cats. SMA was isolated and recorded from the right C-5 phrenic root. Phrenic nerve compound activity was also recorded simultaneously from the contralateral C-5 phrenic root in each preparation. With the vagus nerve intact, phrenic SMA significantly decreased to 91.5, 76.2, 45.3, and 21.6% of control when PEEP was loaded to levels of 2.5, 5, 7.5, and 10 cm H2O, respectively. Compound activity also decreased. Moreover, not only phrenic single postinspiratory inspiratory activity (PIIA) but also the ratio of PIIA to the inspiratory SMA significantly decreased with increments of PEEP. In contrast, with the vagus nerve sectioned, phrenic nerve activity, including PIIA, remained unchanged when loading PEEP. These results suggest that 1) SMA might be inhibited in lung volume-dependent manner by PEEP, and that 2) the vagal nerve afferents are essential for the inhibition of SMA by PEEP. PMID- 8418747 TI - Onset of the nondepolarizing neuromuscular block in humans: quantitative aspects. AB - The purpose of our study was a unified description of the time course for the onset of the neuromuscular block produced by different muscle relaxants after bolus intravenous injections. The environment of the receptors on the motor end plate was assumed to be a part of the interstitial space of the muscle. A unified consideration of four muscle relaxants was accomplished by expressing the concentrations of each relaxant in the interstitial space as multiples of the relaxant's dissociation constant. A flow-limited model was developed to describe the time course of the relaxant's concentration in the interstitial space as a function of its plasma concentration, the plasma flow to the muscle, and the volume of the interstitial space. The results show that those relaxants whose plasma concentrations decrease rapidly achieve an earlier (4-8 min), but relatively lower, peak concentration in the interstitial space. The relaxants with more sustained plasma concentrations reach the peak concentration later, 9 16 min after the bolus intravenous injection. The model allows the interpretation of several observations encountered with the clinical use of the muscle relaxants. PMID- 8418748 TI - Work hours of residents in seven anesthesiology training programs. AB - Medical educators and credentialing organizations recently have called attention to the long hours that some house staff are required to spend in the hospital during training. To determine the average duration of in-hospital work hours of anesthesiology residents, 148 residents at seven, university-affiliated training programs kept daily logs of their activities for one week. Residents in clinical anesthesia years 1, 2, and 3 spent an average of 66, 65, and 64 hours per week, respectively, in the hospital with a range of 43 to 104 hours per week. Although there was not a difference in in-hospital work time among years of training, there was a statistical difference between two of the seven programs studied. The largest portion of the in-hospital time was devoted to patient care activities in the operating room. Residents had time for educational activities, conferences, and reading while in the hospital. The overall work hours of the residents in the anesthesiology training programs included in this survey appeared to be within current guidelines. PMID- 8418749 TI - Zenker's diverticulum. Surgical evolution, perioperative implications. PMID- 8418750 TI - States grapple with HIV/AIDS issues. PMID- 8418751 TI - Special Committee on HIV discusses AIDS, Committee goals. PMID- 8418752 TI - Video technology. PMID- 8418753 TI - Project 2000 final report: a work plan for the future. PMID- 8418754 TI - The effects of n-3 fatty acids on atherosclerosis and the vascular response to injury. AB - The potentially beneficial effects of dietary fatty acids on vascular disease continues to generate intense clinical and investigative interest throughout the world. In particular, attention has focused on the highly unsaturated n-3 fatty acids found in marine and certain plant oils in the prevention of atherosclerosis and thrombosis. This article will review the impact of these fatty acids on vascular disease in epidemiologic studies, in animal models of atherosclerosis and vascular injury, and in human clinical trials of restenosis following angioplasty. Some of the more specific cellular effects of n-3 fatty acids will also be discussed. The data accumulated in the last 15 years suggest that additional prospective clinical trials of n-3 fatty acids in the prevention or treatment of vascular disease are warranted in humans. PMID- 8418755 TI - A clinical laboratory information systems survey. A challenge for the decade. AB - In 1990, the College of American Pathologists Informatics Committee surveyed 14,785 laboratorians for their experiences with a clinical laboratory information system. A 16.25% response rate was achieved, representing 2402 questionnaires that were analyzed. Despite the perceived satisfaction of the clinical laboratory information system users with more expensive systems, no economy of scale was demonstrated with increasing system cost through either laboratory staff reduction or increased number of specimens per day. The strongest predictors of system satisfaction were (1) vendor success measured by number of installations and (2) a selection process that involved the pathologist/laboratory director and included a formal request for proposal. The need for integration of clinical laboratory information systems with hospital information systems, as well as the universal adoption of standard productivity terminology, including work load units, was evident. PMID- 8418756 TI - Physician goals and laboratory test turnaround times. A College of American Pathologists Q-Probes study of 2763 clinicians and 722 institutions. AB - Laboratory test turnaround times (TATs) for emergency department patients were studied in 722 institutions using Q-Probes, a quality improvement program of the College of American Pathologists. The medians of the TATs required by 2763 clinicians were 10 minutes for PO2, 20 minutes for hemoglobin, and 30 minutes for potassium and glucose measurements. Surgeons had the shortest TAT requirements for hemoglobin, potassium, and glucose measurements, whereas emergency department physicians had the shortest requirements for PO2. The measured TATs of most hemoglobin and potassium determinations did not meet clinician goals. In contrast to laboratorians, the majority of clinicians defined a TAT start time as test ordering, and a TAT ending time as result reporting. We recommend laboratorians and clinicians mutually agree on the definition of TAT, jointly develop timeliness goals, and together improve TAT performance to fulfill these goals. PMID- 8418757 TI - Molecular genetics and medicine. A call for papers. PMID- 8418758 TI - Does feedback reduce inappropriate test ordering? PMID- 8418759 TI - A feedback system for reducing excessive laboratory tests. AB - At the James A. Haley Veterans Hospital in Tampa, Fla, a program has been implemented to reduce the amount of potentially excessive laboratory testing. The major program components are a set of test frequency guidelines and a system of feedback to resident physicians that compares their test ordering patterns against the predetermined guidelines. The guidelines are analyte specific and differentiate between normal and abnormal test values reported during 1-day and 7 day time periods. The feedback process includes both systematic reporting of objective data and individual and group education and counseling sessions related to the appropriate use of laboratory tests. A reduction in the percentage of tests that fell outside the guidelines (outliers) was achieved following implementation of the program. PMID- 8418760 TI - Surgical pathology report in the era of desktop publishing. AB - Since it is believed that "a picture is worth a thousand words," incorporation of computer-generated line art was used as a adjunct to gross description in surgical pathology reporting in selected cases. The lack of an integrated software program was overcome by using commercially available graphic and word processing software. A library of drawings was developed over the last few years. Most time-consuming is the development of templates and the graphic library. With some effort it is possible to integrate graphics of high quality into surgical pathology reports. PMID- 8418761 TI - Juvenile rhabdomyoma. An intermediate form of skeletal muscle tumor in children. AB - Two tumors of the buccal soft tissues in children with rhabdomyomatous features are described and further characterized by immunohistochemical studies in both cases and by electron microscopy in one case. Discrete microscopic nodules of elongated, uniform spindle cells with readily identifiable cytoplasmic cross striations replaced existing normal skeletal muscle. In contrast to fetal rhabdomyoma and embryonal rhabdomyosarcoma, there were no immature mesenchymal cells, nor were there individual rhabdomyomatous cells with short, tapered cytoplasmic processes and overtly malignant cytologic features, including mitotic activity. Following excision, one child remains well 46 months later and the other is doing well 7 months after surgery. Some confusion has been created in the literature by the introduction of the terms cellular rhabdomyoma and myxoid fetal rhabdomyoma. We propose that the so-called cellular fetal rhabdomyoma is distinct from the classic fetal rhabdomyoma and may represent the more differentiated juvenile rhabdomyoma. PMID- 8418762 TI - Measurement of factor VIII. A potential risk factor for vascular disease. AB - Factor VIII is a critical component of the coagulation system that is necessary for activation of factor X by factor IXa. Decreased levels of factor VIII are known to be associated with an increased risk of bleeding. It has been suggested that increased levels of factor VIII may be associated with thrombosis and atherosclerosis. Review of published studies indicates that the relationship between an increase in the level of factor VIII and atherosclerosis remains unclear at this time. While there is evidence that patients with atherosclerosis may have an increased levels of factor VIII, it is unclear if this is a primary or secondary event and whether such an increase is predictive of clinical outcome. The interpretation of epidemiologic studies is made more difficult by the current status of clinical laboratory assays for factor VIII activity. Interlaboratory proficiency surveys continue to show relatively high interlaboratory imprecision for these assays. These surveys have documented that a number of laboratories still use suboptimal methods for factor VIII assays and that use of these suboptimal methods is associated with poor performance. Improvement in factor VIII assay performance may be necessary before the role of factor VIII in atherosclerosis can be clearly defined. PMID- 8418763 TI - Factor VII assays. AB - Several prospective and cross-sectional studies have proposed that an association exists between elevated coagulation factor VII levels and cardiovascular disease. Not all of these studies used the same method to assess the factor VII levels. Although the most common method is the one-stage factor assay, there are numerous variables in the composition of this assay. Also, factor VII may circulate in plasma in several forms. The relative contribution of each of these forms to the assay result is presently unknown. Future efforts may help to standardize factor VII assays, improve understanding of the influence of the various forms of factor VII, and identify nonfactor VII components that may both affect assay results and be potential indicators for risk of cardiovascular disease. PMID- 8418764 TI - Performance characteristics of fibrinogen assays. Results of the College of American Pathologists Proficiency Testing Program 1988-1991. AB - Plasma fibrinogen concentration appears to be an important risk factor for the development of atherosclerotic cardiovascular disease of a similar magnitude to cholesterol. The quality control of plasma fibrinogen assays has taken on new importance as a consequence of this potential role as an atherosclerotic risk factor. This article reviews the performance characteristics of 40,000 fibrinogen assays comprising the College of American Pathologists Proficiency Testing Program from 1988 through 1991. Instrument and reagent variables both play roles in the poor interlaboratory reproducibility documented by this study. The absence of either a national or international standard for plasma fibrinogen assays has been a major source of reagent variability. The validation and calibration of the College of American Pathologists lyophilized plasma reference preparation for fibrinogen determination is also reported in this study. The availability of this validated reference plasma should markedly improve interlaboratory reproducibility. PMID- 8418765 TI - Sample preparation for plasma measurement of plasminogen activator inhibitor-1 antigen in large population studies. AB - Plasminogen activator inhibitor-1 is important in regulating fibrinolysis and may be an important cardiovascular risk factor. Because of this, there is increased interest in performing plasminogen activator inhibitor-1 assays in large epidemiologic studies. Our aim in this study was to determine the simplest blood collection methods that yield accurate results with our plasminogen activator inhibitor-1 antigen assay. Our results indicate that the following issues are important: (1) since there is a large circadian variation in plasminogen activator inhibitor-1 plasma levels, a target time frame must be established; (2) commercially available citrate collection tubes are adequate, if sample processing is rapid; (3) careful venipuncture is necessary, with freely flowing collection; hemolysis must be avoided; and (4) centrifugation of at least 30,000 g.min is required to avoid platelet contamination. PMID- 8418766 TI - Intestinal leiomyomatosis: a variant of leiomyomatosis peritonealis disseminata? PMID- 8418767 TI - Laboratory markers of coagulation activation. AB - Advances in our understanding of the biochemistry of the hemostatic mechanism have led to the development of sensitive methods for measuring peptides, enzyme inhibitor complexes, or enzymes that are liberated with the activation of the coagulation system in vivo. Studies employing these markers indicate that a biochemical imbalance between procoagulant and anticoagulant mechanisms can be detected in the blood of humans prior to the appearance of thrombotic phenomena. Properly designed prospective studies will be required to determine whether these assay techniques will enable us to identify individuals who are entering a clinically relevant hypercoagulable state and intervene with appropriate therapy prior to the onset of overt thrombotic disease. PMID- 8418768 TI - Antiphospholipid antibodies and thrombosis. A consequence, coincidence, or cause? AB - Antiphospholipid antibodies have been identified as a potential risk factor for both arterial and venous thromboembolic events. Whether these antibodies are causative or a consequence of a previous clinical event remains controversial. A variety of laboratory assays have been introduced to detect the two primary members of the antiphospholipid antibody family: (1) lupus anticoagulants and (2) anticardiolipin antibodies. There is considerable variability in the laboratory tests that have been utilized. A number of potential pathogenic mechanisms have been proposed for antiphospholipid antibodies. These mechanisms involve the interaction of antiphospholipid antibodies with endothelial cells, platelets, and various plasma regulatory proteins. Although venous thromboembolic events are more common, recent evidence suggests a high frequency of arterial events. The cerebral circulation is most commonly involved, followed by involvement of the coronary arteries. An interesting subset of young patients have peripheral vascular disease that often is relatively refractory to surgical treatment with a high incidence of graft occlusion. PMID- 8418769 TI - Antiplatelet therapy in coronary heart disease. Emerging strategies for the treatment and prevention of acute myocardial infarction. AB - Platelets, long recognized for their role in physiological (protective) hemostasis, have been shown conclusively to play an intricate role in both coronary arterial atherogenesis and acute thrombosis. Accordingly, antiplatelet agents have emerged as a prominent feature in treatment strategies designed to prevent and directly confront the progression and thrombotic complications of coronary heart disease in humans. Antiplatelet therapy, used alone or in combination with thrombin antagonists and thrombolytics must be explored with continued enthusiasm to assure growth of this treatment modality. PMID- 8418770 TI - Oral anticoagulant therapy in the chronic phase of myocardial infarction. AB - A causal role of coronary thrombosis in evolving myocardial infarction is now widely accepted. Patients recovering from an acute heart attack have a higher risk of dying, or suffering another infarction, than an age-matched non-coronary population. Thrombosis plays a major part in this additional mortality and morbidity. The use of oral anticoagulants in the secondary prophylaxis of acute myocardial infarction is controversial. This article reviews the role of oral anticoagulant drugs in the prevention of secondary events in patients who have survived a heart attack. PMID- 8418771 TI - Gastric syphilis mimicking linitis plastica. AB - The decreased incidence of gastric syphilis has made its clinical presentation less widely appreciated. A 61-year-old man suffering from epigastric pain, nausea, and vomiting had an initial diagnosis of gastric carcinoma; the pathologic diagnosis was equivocal. Eventually, gastric syphilis was diagnosed. In the context of the case described below, positive serologic findings in a relatively young adult should raise the suspicion of gastric syphilis. Carcinoma must be ruled out, lest the patient lose valuable time while being treated for syphilis. PMID- 8418772 TI - Laparoscopic jejunostomy using T-fasteners as retractors and anchors. AB - We developed a new technique for performing laparoscopic jejunostomies using T fasteners to secure the jejunum to the abdominal wall. The proximal jejunum is identified with laparoscopy. Four T-fasteners are introduced percutaneously into the jejunal lumen for retraction, and an 8F catheter is inserted through a peel away introducer via a J-wire. The jejunum is drawn up against the abdominal wall by pulling on the T-fasteners. Tube placement is checked with laparoscopy and roentgenography. We performed laparoscopic jejunostomies in five patients using this method, and the results were excellent. Jejunostomies can be performed safely, easily, and reliably this way. PMID- 8418773 TI - The quality of surgery: statistical vs incidental approaches. PMID- 8418774 TI - Frederick Douglass Stubbs Lecture. 'Veritas, dogma, and numbers'. PMID- 8418775 TI - Modulation of macrophage membrane phospholipids by n-3 polyunsaturated fatty acids increases interleukin 1 release and prevents suppression of cellular immunity following hemorrhagic shock. AB - Studies have suggested that the significant suppression of cellular immunity following hemorrhage may be due to an increased release of prostaglandin E2 (PGE2) by macrophages. Since diets high in n-3 polyunsaturated fatty acids decrease PGE2 synthesis, we assessed whether hemorrhage-induced immunosuppression could be prevented by dietary manipulation. C3H/HeN mice were fed for 3 weeks with fat sources derived from corn oil, safflower oil, or fish oil, then bled to a mean blood pressure of 35 mm Hg maintained for 60 minutes. Following this, the animals were adequately resuscitated with fluids and killed 24 hours later. In the corn oil and safflower oil groups, hemorrhage resulted in a significant increase in PGE2 release by peritoneal macrophages, a marked suppression of peritoneal macrophage antigen presentation capacity, interleukin 1 release, splenocyte proliferation, and interleukin 2 secretion compared with shams. However, feeding mice with fish oil for 3 weeks prior to hemorrhage prevented the rise in PGE2 release and maintained normal macrophage and splenocyte functions following hemorrhage. Thus, the elevated release of PGE2 by peritoneal macrophages plays a pivotal role in hemorrhage-induced immunosuppression. Moreover, diets high in n-3 polyunsaturated fatty acids may offer a new therapeutic approach for preventing posthemorrhage immunosuppression and increased mortality from sepsis. PMID- 8418776 TI - Induction of hepatocyte lipopolysaccharide binding protein in models of sepsis and the acute-phase response. AB - Lipopolysaccharide binding protein (LBP) is a serum glycoprotein that complexes with lipopolysaccharide (LPS) to facilitate macrophage response to endotoxin. To determine the conditions that stimulate LBP production in vivo, we measured the induction of LBP in models of inflammation produced by LPS, Corynebacterium parvum, and turpentine injection. Plasma aspartate aminotransferase and alanine aminotransferase concentrations and hepatocyte fibrinogen synthesis were elevated in all models. Northern blot analysis revealed 17-, 14-, and 20-fold upregulation of hepatocyte LBP mRNA following treatment with LPS, C parvum, and turpentine, respectively. Peritoneal macrophage interleukin 6 and tumor necrosis factor production following endotoxin stimulation was augmented by cultured hepatocyte supernatants, suggesting increased LBP synthesis in these groups. The results show that LBP mRNA is induced during hepatic inflammation and suggest that LBP is an acute-phase protein important in regulating the in vivo response to endotoxin. PMID- 8418777 TI - Glutathione depletion in rats impairs T-cell and macrophage immune function. AB - Critical illness is associated with both immunosuppression and glutathione deficiency. We determined if in vivo depletion of glutathione would adversely affect immune status. Rats with normal glutathione levels and those with glutathione stores depleted by diethyl maleate underwent analysis of splenocyte function and mesenteric lymph node lymphocyte function. Lymphocytes of the spleen and mesenteric lymph nodes were tested for concanavalin A proliferative response and interleukin 2 production. Tumor necrosis factor and interleukin 6 secretion by splenic adherent cells was also measured. Glutathione-depleted animals had significantly decreased lymphocyte proliferation and decreased production of tumor necrosis factor and interleukin 6 but unaltered interleukin 2 production. These findings indicate that in vivo glutathione deficiency impairs macrophage and T-cell function. Because glutathione depletion may occur in sepsis, trauma, and shock, treatments that help maintain glutathione levels may enhance immunocompetence and thus improve the ability of patients to recover from critical illness. PMID- 8418778 TI - The risk of exposure of third-year surgical clerks to human immunodeficiency virus in the operating room. AB - The exposure of third-year medical students to blood and blood products in the operating room was assessed with a questionnaire distributed at the end of their clerkship in surgery. Sixty-six (68%) of ninety-seven students reported having been exposed to blood in the operating room during their 3-month rotation in surgery. During the year there was a decrease in the exposure rate that correlated with the students' knowledge of universal precautions (r = .96). Consistent with this observation was a significant decrease in the exposure rate from the first quarter of the year to the last quarter (88% vs 56% of the students). Of the 32 students stuck or cut in the operating room, 21 (66%) were injured by the surgeon. These data underscore the risk to medical students during their clerkships and the important role that universal precautions had in their protection. PMID- 8418779 TI - Impaired polymorphonuclear leukocyte anticandidal function in injured adults with elevated Candida antigen titers. AB - Per protocol, adults with an Injury Severity Score of 18 or greater underwent Candida antigen titer measurements weekly. If titers were 1:4 or greater, neutrophil function against Candida albicans was determined with use of a tritiated glucose incorporation assay, and polymorphonuclear leukocytes obtained from healthy blood donors were studied concurrently for comparison. Polymorphonuclear leukocytes from healthy blood donors and injured patients with elevated titers were able to inhibit C albicans growth in a dose-dependent fashion. Polymorphonuclear leukocytes from injured patients with elevated titers had a significantly depressed ability to inhibit Calbicans growth compared with those from healthy blood donors at all effector cell-to-target cell ratios tested. Cytokine-treated polymorphonuclear leukocytes from healthy blood donors and injured patients with elevated Candida antigen titers demonstrated significantly improved anticandidal activity at all ratios of polymorphonuclear leukocytes-to-Candida. Granulocyte macrophage-colony stimulating factor was the most potent cytokine at reconstituting polymorphonuclear leukocyte function, followed by interferon gamma and interleukin 8. In conclusion, an elevated Candida antigen titer in injured adults is associated with impaired polymorphonuclear leukocyte antifungal activity. This depressed activity can be reconstituted by the addition of cytokine. PMID- 8418780 TI - Insulinlike growth factor 1 (IGF-1) reduces gut atrophy and bacterial translocation after severe burn injury. AB - Bacterial translocation after severe burns is associated with gut mucosal atrophy and increased mucosal permeability. Insulinlike growth factor 1 (IGF-1) levels are low after trauma and do not respond to growth hormone treatment. Since IGF-1 receptors have been demonstrated in gut mucosa, we proposed that treatment with IGF-1 would reduce mucosal atrophy and bacterial translocation. Rats received 50% total body surface area full-thickness burn or sham burn. They were treated with a continuous, subcutaneous infusion of either IGF-1 (approximately 3 mg/kg per day) or placebo (0.01 mol of acetate) for up to 5 days after receiving the burn. The mesenteric lymph node and liver were cultured for gram-negative bacteria. The small intestinal mucosa was scraped, weighed, and analyzed for DNA and protein content. Treatment with IGF-1 improved body weight, spleen weight, and gut mucosal weight. It stimulated mucosal DNA and protein content and reduced the incidence of bacterial translocation to the mesenteric lymph node from 89% to 30%. Insulinlike growth factor may reduce gut barrier failure by decreasing mucosal atrophy and subsequent barrier failure. In addition to its general anabolic effects, recombinant human IGF-1 may improve gut mucosal function and reduce infectious morbidity in severely traumatized or septic patients by reducing gut atrophy and reducing bacterial translocation. PMID- 8418781 TI - Prospective alterations in therapy for penetrating abdominal trauma. AB - In a double-blind, randomized study, 170 patients with traumatic perforation of the gastrointestinal tract were administered an advanced-generation cephalosporin. Patients were divided into infection risk groups (< or = 40%, low; 40% to 70%, mid; and > 70%, high) at surgical closure using a logistic regression formula based on four proved risk factors--age, blood replacement, ostomy, and the number of organs injured. Patients in the low group received 2 days of antibiotic therapy; those in the mid to high group received 5 days of antibiotic therapy. Those patients in the low to mid group had primary wound closure; those in the high group had their wounds packed open and closed later. Most of the patients (144 [85%]) were in the low group. Their major and minor infection rates (10% and 12%, respectively) were not significantly different from 145 historic control subjects receiving 5 days of antibiotic therapy (9% major; 14% minor). Patients in the mid to high group showed a greater incidence of major infections (46%) but a similar incidence of minor infections (12%). The results indicate that risk factors can be used to identify low-risk patients who require only short-term antibiotic therapy and primary wound closure. The remaining patients are at greater risk for infection despite prolonged antibiotic therapy and delayed wound closure. PMID- 8418782 TI - The relationship between platelet count, sepsis, and survival in pediatric burn patients. AB - Early identification of sepsis can be difficult in severe burns because of the systemic changes that routinely accompany these burns. This review examined the value of a falling platelet count in predicting the development of sepsis. Thirty two pediatric patients who sustained lethal burn injuries were compared with 32 patients with similar burns who survived. Daily platelet count was evaluated in conjunction with clinical course. Thirty-one of the 32 non-survivors developed a platelet count less than 0.1 x 10(12)/L. Only 10 of the survivors had a similar occurrence. Platelet count decline preceded other signs of sepsis in all cases. A platelet count below 0.1 x 10(12)/L for more than 4 days was uniformly associated with death. All patients who died succumbed to multisystem organ failure, consistent with sepsis. These results emphasize platelet count as an independent predictor of sepsis and death. PMID- 8418783 TI - Polymorphonuclear cell-mediated vascular injury in anergic surgical patients. AB - We examined the responses of primed polymorphonuclear neutrophils (PMNs) adhered to vascular endothelium, which can lead to endothelial cell damage as a mechanism of the capillary leak syndrome, the main cause of death in anergic patients. We tested PMNs from (1) preoperative reactive patients, (2) preoperative anergic patients, (3) anergic patients in the surgical intensive care unit, and (4) healthy controls for in vitro adherence and cytotoxicity on cultured human vein endothelial cells. Adherence of PMNs was 12.9% +/- 3.9% in preoperative anergic patients and 13.1% +/- 3.2% in anergic patients in the surgical intensive care unit compared with 9.0% +/- 2.1% in preoperative reactive patients (P < .05). Cytotoxicity was 6.0% +/- 2.8% in preoperative reactive patients, 13.7% +/- 4.1% in preoperative anergic patients, and 14.3% +/- 4.6% in anergic patients in the surgical intensive care unit. The PMNs from preoperative anergic patients were more cytotoxic against human vein endothelial cells when stimulated by Staphylococcus epidermidis or formyl-methionyleucylphenylalanine. We conclude that PMNs from anergic surgical patients adhere more to endothelial cells and can produce increased cytotoxicity that may lead to detrimental results. PMID- 8418784 TI - Anti-tumor necrosis factor antibody reduces mortality in the presence of antibiotic-induced tumor necrosis factor release. AB - The systemic tumor necrosis factor (TNF) response has been extensively studied during infection. In addition, antibiotics that cause cell-wall lysis have been associated with endotoxinemia and, therefore, could trigger TNF release. We studied the effects of pretreatment with cefoxitin and/or anti-TNF antibody on mortality and early (90 minutes) and delayed (6 hours) serum TNF levels in a murine model of mixed Escherichia coli/Bacteroides fragilis peritonitis. At low and intermediate inocula levels, cefoxitin, but not anti-TNF antibody, prevented death, and low serum TNF levels were noted in all groups. At the highest inoculum level, mortality was uniform in control, cefoxitin, and anti-TNF antibody groups, and a significant elevation in serum TNF levels was seen only at the 6-hour point in animals receiving cefoxitin. The addition of anti-TNF antibody to cefoxitin at this inoculum level abrogated the 6-hour rise in serum TNF levels and reduced mortality to 40%. These results emphasize that the cytokine response in disease is dependent on both the nature of the insult and other forms of therapeutic interventions. PMID- 8418785 TI - Antimicrobial prophylaxis for surgical wounds. Guidelines for clinical care. AB - Prophylactic administration of antibiotics can decrease postoperative morbidity, shorten hospitalization, and reduce the overall costs attributable to infections. Principles of prophylaxis include providing effective levels of antibiotics in the decisive interval, and, in most instances, limiting the course to intraoperative coverage only. Use in The National Research Council clean contaminated operations is appropriate and, in many instances, has been proven beneficial. Antibiotic prophylaxis is also indicated for clean operations, such as those involved with insertion of prosthetic devices, that are associated with low infection risk and high morbidity. Extension of antibiotic prophylaxis to other categories of clean wounds should be limited to patients with two or more risk factors established by criteria in the study of the efficacy of nosocomial infection control (SENIC) because the baseline infection rate in these patients is high enough to justify their use. Cefazolin (or cefoxitin when anaerobic coverage is necessary) remains the mainstay of prophylactic therapy. Selection of an alternate agent should be based on specific contraindications, local infection control surveillance data, and the results of clinical trials. Newer criteria for determining the risk of "site infection" (wound and intracavitary) are in evolution and may lead to modification of these recommendations over the next several years. PMID- 8418786 TI - Role of interleukin 6 and transforming growth factor-beta in the induction of depressed splenocyte responses following sepsis. AB - We examined whether (1) there is an association between elevated circulating levels of transforming growth factor-beta (TGF-beta) and splenocyte dysfunction during sepsis, and (2) administration of monoclonal antibodies to interleukin 6 (an inducer of TGF-beta release) or TGF-beta could ablate these changes. Blood and splenocytes were obtained from C3H/HeN mice at 1, 4, or 24 hours following cecal ligation and puncture or sham operation. Only at 24 hours after cecal ligation and puncture was there an association between elevated blood TGF-beta value and depressed splenocyte interleukin 2 release. Administration of monoclonal antibodies against interleukin 6, but not against TGF-beta (intraperitoneally immediately following cecal ligation and puncture), significantly decreased the blood levels of TGF-beta at 24 hours following cecal ligation and puncture and improved splenocyte interleukin 2 release. Thus, the judicious use of monoclonal antibodies against interleukin 6 may block the subsequent elevation of TGF-beta, thereby attenuating host immunosuppression during sepsis. PMID- 8418787 TI - The activation of bone marrow macrophages 24 hours after thermal injury. AB - We determined the effect of thermal injury on the in vitro production of the immunoactive substances tumor necrosis factor, interleukin 1, prostaglandin E2, and complement component C3 by lipopolysaccharide-stimulated guinea pig bone marrow macrophages and on the cytotoxicity of these cells. Macrophages from burned animals produced different amounts of these mediators compared with unburned animals at certain culture times, suggesting that thermal injury could program the bone marrow cells to respond differently from normal cells to in vitro stimulation with lipopolysaccharide. Also, the macrophages from burned animals displayed greater cytotoxicity towards L929 target cells. These results suggest that there is a complex interaction among cellular secretory products, especially after thermal injury, that may be important in host defense. PMID- 8418788 TI - Synthesis of dengue virus RNA in vitro: initiation and the involvement of proteins NS3 and NS5. AB - An assay for flavivirus RNA-dependent RNA polymerase activity in vitro was established using extracts of Vero cells infected with dengue virus type 2 (DEN 2) or Kunjin virus (KUN). RNA synthesis was initiated on a template of viral replicative form (RF) and RF was converted to the replicative intermediate (RI). The RNA-dependent RNA polymerase complex of DEN-2 utilised either DEN-2 or KUN RF as template, and similarly the KUN polymerase complex utilised either DEN-2 or KUN RF template. In addition, antibodies against the nonstructural proteins NS3 and NS5 inhibited the conversion of RF to RI, indicating that NS3 and NS5 are involved in viral RNA replication. PMID- 8418789 TI - Cloning and nucleotide sequence of the capsid protein and the nuclear inclusion protein (NIb) of potato virus A. AB - The sequence of the 3'-terminal 2597 nucleotides of potato virus A (PVA) genome has been determined from cDNA clones. An open reading frame was identified and potentially encodes a large polyprotein containing 789 amino acid residues. This large open reading frame was found to have a high similarity to the nuclear inclusion protein (NIb) and the capsid protein (CP) genes of several potyviruses. The data suggest that the PVA NIb and CP are products arising from the maturation of the large polyprotein as observed for other potyviruses. Putative cleavage sites corresponded to consensus sequences NIa/NIb and NIb/CP, respectively, of other potyviruses. The NIb (putative RNA polymerase) and CP are expected to be 516 and 269 amino acid residues (M(r) of 58,939 and 30,094), respectively. The non-coding region is 227 nucleotides long, rich in A and U and unlike other viruses. Furthermore, there are two AUG codons in frame in front of the capsid protein gene suggesting an alternative mode for the capsid protein expression. PMID- 8418790 TI - Molecular cloning, sequencing, and expression in Escherichia coli of the potato virus Y cytoplasmic inclusion gene. AB - Complete nucleotide sequences of cytoplasmic inclusion (CI) genes of two strains of potato virus Y (PVY) were determined from six polymerase chain reaction (PCR) amplified cDNA clones. The size of the CI genes of both ordinary (PVY-O) and necrotic strains (PVY-T13) was 1902 nucleotides, with a sequence homology of 83.4%. Comparison of the predicted amino acid sequences showed more than 90% homology. When these were compared with those of other potyviruses, the homology ranged from 53 to 61%. cDNAs of all or a part of the PVY-O CI gene containing an additional initiation codon (ATG) at the 5' end and a stop codon at the 3' end were constructed by PCR amplification and cloned into an Escherichia coli expression vector, pKK 223-3. Complete and truncated PVY-O CI proteins were successfully produced in E. coli as judged by reactivities with PVY-O CI protein specific antiserum. To our knowledge, this is the first report on expression of PVY CI proteins in E. coli. PMID- 8418791 TI - In vitro transcription and replication of the mumps virus genome. AB - By the use of lysolecithin-permealized extracts from mumps virus-infected HeLa cells, we have developed an in vitro system, which not only directed the synthesis of mumps virus mRNAs but also supported replication of the genomic RNA. Furthermore, upon transcription of the P gene, both faithful and edited copies of the P gene were detected by RNase mapping with a riboprobe. Thus this system seems to promote biochemical analyses of underlying mechanisms operative in mumps virus gene expression and replication, including RNA editing. PMID- 8418792 TI - Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenza virus A/WSN. AB - The MDBK-R cell line is a variant of the MDBK cell line, which was derived by three consecutive high multiplicity superinfections of MDBK cells with AWBY-140 virus, a mutant of influenza virus A/WSN (H 1N 1). MDBK-R cells are permissive for productive replication of AWBY-140, but resist lysis by the virus and grew normally without producing infectious virus after replication of the mutant occurred there. By polymerase chain reaction (PCR), we demonstrated nucleotide sequences specific to all the 8 genes of AWBY-140 in MDBK-R cells which had been infected with the mutant at a high multiplicity and subsequently received 25 passages. This suggests that the genes of influenza virus mutant persisted in the dividing host cells for a long time after productive infection, when none of the cells was producing virus. We were also able to amplify the M gene related sequence of the mutant from both poly(A)+ and poly(A)- fractions of the RNA extracted from the cells at 27th passage level by PCR, which suggests that the persisting genes were replicated and transcribed, but we failed to demonstrate any viral protein in the cells by Western blotting. PMID- 8418793 TI - Biopsy results in a kindred with Lafora disease. AB - We studied biopsy results in a kindred with the Lafora form of progressive myoclonic epilepsy. Four members of a family with known consanguinity presented as teenagers with seizures, myoclonus, dementia, and ataxia. After the diagnosis was established by brain biopsy in the first patient, many efforts were made to obtain a tissue diagnosis in the three other patients. Lafora bodies were absent in most of the skin biopsy specimens in three patients and in liver biopsy specimens from two patients. In cases of Lafora disease, where a reasonably certain clinical diagnosis can be established, supported by biopsy proof in some family members, repeated biopsy specimens even at advanced stages of the disease may be negative. These findings suggest that negative skin or liver biopsy specimens in patients with progressive myoclonic epilepsy should not exclude the diagnosis of Lafora disease. PMID- 8418794 TI - Early onset of leuko-araiosis in hypertensive patients. PMID- 8418795 TI - Clinical and magnetic resonance features of the classic and akinetic-rigid variants of Huntington's disease. AB - We studied 32 patients with confirmed Huntington's disease (HD); six (mean age, 31.7 years) had the akinetic-rigid form and 26 (mean age, 46.1 years) had the classic hyperkinetic form. Clinical examination included a count of abnormal involuntary movements, motor self-sufficiency evaluation by the Physical Disability Rating Scale, cognitive function assessment by the Mini-Mental State examination, and a verbal fluency test. Magnetic resonance imaging permitted measurement of bicaudate diameter, a sensitive indicator of caudate atrophy in HD. Patients with the akinetic-rigid form of HD were younger and had earlier disease onset than those with the classic form of HD. All patients with akinetic rigid HD (group 1) had striatal hyperintensity on T2-weighted magnetic resonance images; seven patients with classic HD (group 2) had a similar abnormality. Groups 1 and 2 were in fact similar in all other respects, except that the number of abnormal involuntary movements was greater in group 2. Groups 1 and 2 together had significantly younger age at onset, lower Mini-Mental State Examination score, more severe motor disability, worse verbal fluency test result, and greater bicaudate diameter than the 19 patients with classic HD without magnetic resonance signal abnormality (group 3) and appear to be a uniform population, distinct from group 3. The abnormalities on magnetic resonance images indicated greater striatal damage in groups 1 and 2, which could be the neuroanatomic substrate of their greater motor and cognitive compromise. PMID- 8418796 TI - Idiopathic, progressive mononeuropathy in young people. AB - We describe six young patients with insidiously progressive, painless weakness in the distribution of a single major lower extremity nerve. No cause could be found despite extensive evaluation, including surgical exploration. At the time of diagnosis, all patients had weakness and three patients had sensory loss. In all cases, electromyography revealed a chronic axonal mononeuropathy without conduction block or focal conduction slowing. Magnetic resonance, computed tomographic, and ultrasound imaging studies did not identify a region of nerve swelling, mass, or compression. At surgical exploration, the nerve appeared atrophic in two patients, indurated in one patient, and normal in two patients. Biopsy specimens obtained from two abnormal nerves revealed either wallerian degeneration or endoneurial fibrosis. The clinical features of these patients comprise an unusual clinical entity with no known cause or treatment. PMID- 8418798 TI - The results of clinical trials in stroke rehabilitation research. AB - RESEARCH PROBLEM AND METHODS: There are currently 1.5 million stroke survivors in the United States. More than half of these individuals have significant residual physical disability and functional impairment. Survivors of stroke constitute the largest group of patients receiving rehabilitation services in this country. We examined existing clinical trials investigating the effectiveness of stroke rehabilitation programs to improve functional outcomes and discharge destination. One hundred twenty-four research reports were initially identified. From this sample, 36 trials meeting selected criteria were evaluated by the methods of meta analysis. RESULTS: A total of 3717 patients participated in the 36 clinical trials included in the meta-analysis. The results revealed a mean d-index of 0.40 +/- 0.33. This effect size index was converted to a U3 value of 65.5, indicating that the average patient receiving a program of focused stroke rehabilitation performed better than approximately 65.5% of those patients in comparison groups (95% confidence interval, 63.6% to 67.3%). The results also revealed a significant interaction between type of research design and method of recording the outcome of a clinical trial. Blind recording of the outcome measure appears to be an essential design characteristic in clinical trials that do not randomize patients to conditions. CONCLUSIONS: Programs of focused stroke rehabilitation may improve functional performance for some patients who have experienced a stroke. The improvement in performance appears related to early initiation of treatment, but not to the duration of intervention. Improvements are also associated with the patient's age and the type of design. Research design should be considered an important moderator variable in planning and interpreting future clinical trials of treatment effectiveness in stroke rehabilitation. PMID- 8418797 TI - Long-term antiepileptic efficacy of vigabatrin in drug-refractory epilepsy in mentally retarded patients. A 5-year follow-up study. AB - The long-term clinical, neurophysiologic, and psychological effects of add-on vigabatrin treatment were evaluated in a group of 36 mentally handicapped patients with drug-refractory epilepsy. After an initial 3-month follow-up period, 15 (42%) of 36 patients had at least a 50% decrease in seizure frequency compared with baseline. After a 2-year follow-up period, nine (25%) of 36 patients retained the initially observed antiepileptic effects of vigabatrin, and after 5 years, eight (22%) of 36 patients did so. Five (33%) of the 15 patients who initially exhibited a favorable antiepileptic response to vigabatrin lost that response during a 5-year follow-up. Partial-onset seizures represented the seizure type best controlled by vigabatrin. Side effects were mostly mild, and plasma levels of other antiepileptic medication remained unchanged. No impairment of psychological performance was observed during vigabatrin treatment compared with baseline. Also, no clear change was observed in the background or epileptiform activity in the electroencephalogram during the study. Our findings suggest that vigabatrin as an add-on therapeutic effectively controls seizures in a subpopulation of patients with severe epilepsy. In addition, the antiepileptic response, if achieved, is long lasting in about half of the patients. PMID- 8418799 TI - Presurgical evaluation of temporal lobe epilepsy using interictal temporal spikes and positron emission tomography. AB - OBJECTIVE: Our goal was to determine the role of fludeoxyglucose F 18-positron emission tomography (18FDG-PET) and interictal temporal spikes in lateralizing the epileptogenic region in patients who (1) were diagnosed as having temporal lobe epilepsy based on clinical symptoms and exclusively temporal interictal spikes and (2) did not have a structural lesion on magnetic resonance imaging. DESIGN: This was a retrospective study of 40 consecutive patients fulfilling the above criteria who underwent 18FDG-PET scanning. A firm electrophysiologic diagnosis and 1 complete year of postsurgical follow-up, where applicable, were required. Outcome measures included surgical outcome and final electrophysiologic diagnosis. RESULTS: Unilateral, interictal temporal spikes (ITS) were present in 33 (82.5%) of 40 patients. Seven patients (17.5%) had bitemporal, independent spikes. Thirty-one (77.5%) of 40 patients had unilateral temporal hypometabolism (TH). Twenty-eight (70%) patients had concordant TH and ITS. One year after surgery, 31 of 33 patients with unilateral ITS were greatly improved; two of five who had bitemporal ITS showed similar improvement. In 28 patients, unilateral TH and unilateral ITS were concordant. The paired result always concurred with the final neurophysiologic assessment. Surgical outcome between patients with 18FDG PET showing unilateral TH (26 of 30 greatly improved) and those not showing unilateral TH (six of eight greatly improved) was not significantly different. CONCLUSION: In temporal lobe epilepsy not associated with a mass lesion, unilateral ITS are reliable lateralizing features and suggest a good surgical outcome. Use of 18FDG-PET provides corroborative lateralizing information but 18FDG-PET that fails to show unilateral TH does not preclude a good surgical outcome. PMID- 8418800 TI - Extrapyramidal signs and other neurologic findings in clinically diagnosed Alzheimer's disease. A community-based study. AB - The association between findings on the neurologic examination and the clinical diagnosis of Alzheimer's disease was investigated among 467 individuals from a geographically defined community population. Participants were selected by stratified random sampling based on their memory performance in a population survey of community residents 65 years of age and older. Each participant underwent a structured medical, psychiatric, neurologic, and neuropsychologic examination. Of the 467 persons examined there were 134 cases of probable Alzheimer's disease and 167 control subjects. Multiple logistic regression analysis was used to estimate the degree to which the presence of each of several neurologic examination findings affected the age- and sex-adjusted relative odds of having clinically diagnosed Alzheimer's disease. The most striking associations with the diagnosis of Alzheimer's disease were seen with various measures of extrapyramidal dysfunction. These increased relative odds were not markedly affected by excluding from the analysis cases with severe cognitive impairment. The results suggest that involvement of the extrapyramidal system is a common finding in Alzheimer's disease. PMID- 8418801 TI - Reliability and usefulness of a new immunochemical assay for Alzheimer's disease. AB - OBJECTIVE: To assess the reliability and usefulness of a new sandwich enzyme linked immunoassay (ALZ-EIA) that detects Alzheimer's disease-associated proteins in the diagnosis of Alzheimer's disease. DESIGN: The reliability of the assay was assessed between two laboratories. Sensitivity and specificity of a diagnostic algorithm based on the results of the ALZ-EIA were determined using the Consortium to Establish a Registry for Alzheimer's Disease neuropathological diagnoses as the "gold standard." SETTING: Autopsy cases were obtained from a teaching hospital with a specialized Alzheimer Disease Diagnostic and Treatment Center. CASES: Brain tissue was selected from 24 cases with dementia and 10 normal controls. MAIN OUTCOME MEASURES: Optical density measurements from the ALZ EIA in the hippocampus and three neocortical regions. RESULTS: A 95% concordance in ALZ-EIA activity was found between the two laboratories, and an 85% concordance was found between ALZ-EIA and the Consortium to Establish a Registry for Alzheimer's Disease diagnoses. Perfect agreement was obtained for "typical" Alzheimer's disease cases (those with plaques and tangles), while discrepancies occurred for "atypical" cases (those with predominantly plaques or tangles). CONCLUSIONS: The ALZ-EIA provides a highly reliable method of assessing neurofibrillary degeneration. Its clinical usefulness as a diagnostic test would be enhanced by the availability of a complementary assay for beta-amyloid. PMID- 8418802 TI - Predictive value of magnetic resonance imaging in temporal lobe epilepsy surgery. AB - The predictive value of magnetic resonance imaging (MRI) was assessed by a prospective study of 34 patients selected for surgical treatment of temporal lobe epilepsy. The MRIs were interpreted using standardized visual diagnostic criteria and the imaging findings were correlated with the surgical outcome. Lateralized MRI abnormalities were found in 25 (74%) of 34 patients. Significant associations were found between either the presence of a restricted foreign-tissue lesion or hippocampal atrophy and an excellent surgical outcome. An abnormal MRI had an 82% predictive value and a normal MRI had a 56% predictive value for surgical success. A history of febrile convulsions and the presence of hippocampal atrophy best predicted outcome (predictive value, 86%). These results suggest that specific MRI findings in candidates for temporal lobe epilepsy surgery are predictive of surgical outcome. The information provided by MRI may be of value for counseling patients prior to surgical intervention. PMID- 8418803 TI - Differential diagnosis in dementia. Principal components analysis of clinical data from a population survey. AB - OBJECTIVE: To reduce all the clinical data, collected from an unselected group of subjects, to a small set of factors and to see how these factors correspond to standard clinical diagnosis of dementing disorders. DESIGN: Population survey. SETTING: General community: elderly older than 74 years, from an area in Stockholm, Sweden. SUBJECTS: Population-based sample including (1) all the screened positive subjects using the Mini-Mental State examination; and (2) a random sample of the screened negative subjects, matched by age and sex. A clinical examination and an informant interview were carried out. Cases were identified using Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition diagnostic criteria for Alzheimer's disease (AD) and other dementias. MAIN OUTCOME MEASURE: Independently from the clinical diagnosis, a principal components factor analysis was carried out to investigate groupings among the clinical data (factors). Factor scores, calculated as a weighted sum of the symptom variables and converted to a standard score form. RESULTS: Four major factors were found: cognitive impairment, cerebrovascular disease, disturbed behavior, and depressive symptoms. The comparison of these factors with the clinical diagnoses showed that (1) the cognitive impairment factor discriminated demented cases from nondemented; (2) the cerebrovascular disease factor discriminated vascular dementia from AD cases and nondemented; (3) the disturbed behavior factor discriminated AD cases from vascular dementia cases and nondemented, indicating behavioral changes characteristic of AD. CONCLUSIONS: This finding, if replicated, would have implications for the construction of diagnostic criteria for AD. PMID- 8418804 TI - Idiopathic intracranial hypertension (pseudotumor cerebri). Descriptive epidemiology in Rochester, Minn, 1976 to 1990. AB - The medical records-linkage system of the Mayo Clinic was used to identify cases of idiopathic intracranial hypertension in the 15-year period, 1976 through 1990, among the population of Rochester, Minn. Nine patients (eight women and one man) were identified, corresponding to an average annual age-adjusted incidence rate per 100,000 of 0.9 for the total and 1.6 for the female population. In females aged 15 to 44 years, idiopathic intracranial hypertension occurred at a rate of 3.3 per 100,000 per year; for those defined as obese (body mass index > 26), the rate rose to 7.9. Median follow-up was 2.7 years (range, 5 months to 15 years). Three of 18 eyes developed visual impairment; this was mild in all cases. PMID- 8418805 TI - On the preservation of syntax in Alzheimer's disease. Evidence from written sentences. AB - We examined the syntactic complexity of single written sentences elicited from 368 adults undergoing examination for possible Alzheimer's disease. The clause length and composition of the sentences varied with the severity of dementia. The nondemented adults' sentences contained more propositions, main and secondary verbs, and conjunctions than those produced by the mildly and moderately demented adults. Sentence length in clauses, propositional content, and the use of conjunctions and main and secondary verbs discriminated among stages of the severity of the disease. The present results suggest that, while dementia severity affects written linguistic output, such output is, nonetheless, grammatic and coherent. PMID- 8418806 TI - Cognitive functioning after common whiplash. A controlled follow-up study. AB - A random sample of 98 patients with common whiplash was examined early after trauma (mean +/- SD, 7.3 +/- 3.9 days) and again 6 months later. Cognitive functioning was assessed in conjunction with complaints, pain intensity, well being, subjective cognitive impairment, neuroticism, and medication. At 6 months, 67 patients had fully recovered (asymptomatic group), while 31 were still symptomatic (symptomatic group). Symptomatic patients who were older at baseline, had a greater variety of symptoms, higher neck pain intensity, and greater subjective cognitive impairment. At baseline, both groups scored poorly on tests requiring complex attentional processing. All neuropsychological functions improved to normal at 6 months in both groups. This improvement cannot be explained by a practice effect, as shown by the results of normal volunteers. The symptomatic group showed delayed recovery regarding complex attentional functioning, which may be related to adverse effects of medication. PMID- 8418807 TI - Memory evaluation in Alzheimer's disease. Caregivers' appraisals and objective testing. AB - OBJECTIVES: To evaluate if caregivers are reliable informants concerning memory deficits in patients with Alzheimer's disease (AD). DESIGN: Responses of caregivers of patients with probable AD and responses of healthy control subjects on a standardized memory questionnaire were compared with objective measures of cognition (Consortium to Establish a Registry for Alzheimer's Disease neuropsychological battery) and with clinical estimates of activities of daily living, depression, and psychopathology (Consortium to Establish a Registry for Alzheimer's Disease [CERAD] clinical assessment battery) using the Self-report Memory Questionnaire. SETTING: A federally funded AD research center. SUBJECTS: The referred sample included 117 patients with probable AD, their informants, and 41 healthy control subjects age-matched to the patients. Patients and control subjects were between the ages of 58 and 85 years, had between 9 and 19 years of education, and were in good health. EXCLUSIONS: Patients who did not meet NINCDS ADRDA criteria of probable AD. MAIN OUTCOME MEASURE: The optimal number of questionnaire items yielding the best combination of sensitivity and specificity. RESULTS: An abbreviated version of the scale, renamed the Short-Memory Questionnaire, had excellent specificity and sensitivity for identifying dementia. Positive and negative predictive values were 63.5% and near 100%, respectively. The Short-Memory Questionnaire showed good reliability, internal consistency, and external validity. Caregiver appraisals of memory deficits significantly correlated with objective measures of memory and also with generalized cognitive dysfunction. CONCLUSIONS: Caregivers of patients with AD are reliable informants of their relatives' deficits. The Short-Memory Questionnaire is an easily administered, informant-based scale that may be useful in clinical settings or epidemiologic studies to screen out persons with memory difficulties. PMID- 8418808 TI - Time course of cerebral blood flow velocity in central nervous system infections. A transcranial Doppler sonography study. AB - In a 3-year period, 110 patients with central nervous system infections of various causes were examined serially by means of transcranial Doppler sonography. In viral-induced infections, no changes of flow velocity in basal cerebral arteries were seen, whereas in bacterial meningitis, a significant increase of blood flow velocity in the middle cerebral artery was recorded. Its extent was mainly associated with the type of the infectious agent, most frequently observed in pneumococcal meningitis (77%). The increase was up to 100% of the baseline values and was reversible in all cases. All patients were offered full-scale neurointensive care, and all subjects with bacterial meningitis were fully heparinized. PMID- 8418809 TI - Molecular cloning of a novel mRNA sequence expressed in cleavage stage mouse embryos. AB - In an approach to study genes transcribed during early mouse development, a cDNA library was constructed from poly(A) RNA isolated from the 8-cell morula. The cDNA library was differentially screened with labelled cDNA probes synthesized on poly(A) RNA isolated from the 8-cell morula or unfertilized eggs. Six clones which increased in abundance in the 8-cell morula were selected and further analyzed. Sequencing analyses showed that some of these clones corresponded to RNA transcripts from B1 and B2 repetitive sequences, as well as mRNA for cytochrome C oxidase I and NADH dehydrogenase III derived from the mitochondrial genome. One clone was not identical to any known sequences. The unidentified sequence (MO25) was found at low levels in the unfertilized egg, but increased at the 2-cell stage. The predicted amino acid sequence revealed that the MO25 gene may encode a Ca2+ binding protein. PMID- 8418810 TI - Factor in urinary extracts from pregnant women that inhibit mouse oocyte maturation in vitro. AB - Mouse oocyte maturation inhibitory factors, on the basis of inhibitory activity of spontaneous germinal vesicle breakdown (GVBD) of denuded mouse oocytes in culture, were extracted and partially purified by reversed-phase resin adsorption and Sephadex G-100 and G-50 column chromatographies from the urine of pregnant women. Denuded oocytes obtained from ovaries of ICR mice underwent spontaneous GVBD by cultivation for 3 h in modified Krebs-Ringer's buffered solution, while this spontaneous GVBD was found to be inhibited by adding the final preparation (U-D-4) of urine. The inhibition was dose dependent, ranging from 0.6 to 10 micrograms protein/ml medium. Oocytes treated with U-D-4 and resuspended in control medium resumed GVBD. The molecular mass of U-D-4 was estimated to be less than 2,000 Da with gel filtration. Ether treatment failed to extract inhibitory factor(s) from U-D-4 and pepsin treatment inactivated U-D-4, indicating that inhibitory factor(s) in U-D-4 are peptide-like substances. The inhibitory effect of U-D-4 on spontaneous GVBD was partially reversed in the presence of naloxone, a potent opioid antagonist. U-D-4s obtained from urine samples of pregnant women, nonpregnant women, and men showed the inhibitory effect on spontaneous GVBD; however, the activity of U-D-4 obtained from pregnancy urine was significantly more potent than those of the other urine samples. PMID- 8418812 TI - Region-specific variation of gene expression in the human epididymis as revealed by in situ hybridization with tissue-specific cDNAs. AB - Three tissue-specific gene probes that had been isolated by differential screening from a human epididymal cDNA library--HE1, HE2, and HE5--were employed to investigate regional specializations in the human epididymis. All 3 cDNAs were derived from major transcripts of the epithelial cells lining the epididymal duct. Each mRNA species, however, exhibited a discrete longitudinal pattern of hybridization with maxima in different regions of this organ, suggesting regional specializations of gene expression. The HE5 mRNA, which was recently shown to encode the peptide backbone of the human leukocyte differentiation antigen CDw52, showed maximum levels in the distal corpus epididymidis and in the vas deferens, whereas the HE2 mRNA was found predominantly in the caput and proximal corpus sections of the epididymis. HE1 mRNA was found in high amounts in all parts of the epididymis, displaying a 2-peak expression pattern with maxima in the distal caput and distal corpus of the epididymal duct. PMID- 8418811 TI - Characterization of a cDNA clone coding for human testis membrane cofactor protein (MCP, CD46). AB - Membrane cofactor protein (MCP) is a complement regulatory protein that acts as a cofactor for the cleavage of C3b and C4b by the serine protease factor I. We have previously reported the characterization of a functional MCP molecule on the acrosomal membrane. This protein migrated as a single band with a molecular weight of 40,000 Da, which is 10,000-20,000 Da smaller than the known MCP molecules, and is devoid of N- and O-linked sugars. We have proposed that the difference in molecular weight resulted from the lack of sugars. To investigate if this is due to the absence of glycosylation sites, we have characterized a cDNA clone from a human testis cDNA library. This cDNA corresponds to a peculiar MCP form previously described, which is characterized by the presence of the serine/threonine/proline-rich exon C (STPC) and the cytoplasmic tail known as CYT2, and we conclude that the absence of mature oligosaccharide of the sperm MCP cannot be totally attributed to a defect of N- and O-glycosylation sequences but rather reflects an alteration of the mechanisms of glycosylation in spermatozoa. The presence of functional MCP on the acrosomal membrane, as well as the other complement regulatory protein, decay-accelerating factor, strongly suggests that these proteins may act concomitantly to protect the acrosome-reacted spermatozoa from the attack of the complement present in the female genital tract. PMID- 8418813 TI - Embryo sex selection by a rat male-specific antibody and the cytogenetic and developmental confirmation in cattle embryos. AB - Embryos of mouse, rabbit, goat, sheep, and cattle were separated into 2 groups on the basis of their morphology when incubated with a male-specific antibody (qualified here as the H-Y antibody) prepared from newborn rat testis. When morula-stage embryos were cultured in the presence of this H-Y antibody, the development of roughly one half of the embryos was arrested at that stage, whereas the other half continued to develop to the blastocyst stage. The developmentally arrested group of embryos resumed their development into blastocysts when cultured in antibody-free medium. Eighty to 90% of cattle embryos whose development was unaffected by the antibody were shown to possess a female karyotype (XX), and close to 80% of those embryos whose development was arrested possessed a male karyotype (XY). Cattle embryos whose sex had been presumptively identified by development in the presence of the H-Y antibody were cryopreserved and transferred, and the sex of the calves was examined. The overt sex of the young born from sexed embryos was found to be the same as that determined by chromosomal analysis. PMID- 8418814 TI - Chromatin and microtubule organization in the first cell cycle in rabbit parthenotes and nuclear transplant embryos. AB - Artificial activation and nuclear transfer in rabbit oocytes have been used in past years in an attempt to develop viable techniques for cloning in cattle. The procedures established in our laboratory, using the rabbit as a model, consistently lead to high rates of development to the blastocyst stage. However, the rate of embryos developing to term is considerably lower. In the present study, we undertook a detailed immunocytochemical study of the patterns of both microtubules and chromatin during the first cell cycle of electrical pulse activated oocytes and of nuclear transfer embryos. Our goal was to investigate the responses of the cell to the different stimuli applied and to establish the sequence of events leading to first cleavage in the absence of normal fertilization. Our results show that, in both electrically activated oocytes and nuclear transfer embryos, although the initial development patterns are rather unusual, embryos become synchronized at the time of the formation of a pronuclear like structure, and then organize metaphase spindles and cleave. These spindles consistently present small defects, suggesting that problems in the formation of the mitotic apparatus during the first cell cycle may have a long-term effect leading to embryo mortality. PMID- 8418815 TI - Development of androgenetic mouse embryos produced by in vitro fertilization of enucleated oocytes. AB - Enucleated mouse oocytes were successfully fertilized in vitro, and the resultant androgenetic eggs developed to the blastocyst stage. The proportion of enucleated oocytes fertilized in vitro was high (87-99%) at sperm concentrations ranging from 10-100 x 10(4)/ml. At high sperm concentrations (100-1,000 x 10(4)), 35-45% of the fertilized eggs resulted in heterozygous bispermic androgenones. The proportion of hemizygous haploid and heterozygous diploid androgenones developing to blastocysts was 11% and 43%, respectively. Hemizygous diploidization, however, showed no positive effect on development. These results clearly show that the procedure reported here is efficient and reliable for the production of androgenetic eggs. PMID- 8418816 TI - Forskolin and the meiosis inducing substance synergistically initiate meiosis in fetal male germ cells. AB - We have shown that Meiosis Inducing Substance (MIS) and forskolin synergistically and dose dependently induce meiosis in germ cells of cultured fetal mouse testes. We used a bioassay which consists of fetal mouse testes and ovaries cultured for 6 days. In this study MIS media are spent culture media from 24 hour cultures of minced adult mouse testes. In the bioassay one gonad of each fetus is cultured either in MIS medium, in control medium with forskolin, or in MIS medium with forskolin. The other gonad serves as the control and is cultured in control medium. After culture the gonads are fixed, squashed, and DNA-stained. In these preparations germ cells and somatic cells can be distinguished, and the number of germ cells in the different stages of meiosis is counted as is the number of somatic cells in mitosis. MIS activity is defined to be present in a medium when meiosis is induced in male germ cells during culture. We found that MIS media as well as forskolin induced meiosis in fetal male germ cells in a dose-dependent manner. In addition, MIS media and forskolin acted synergistically by inducing meiosis. Female germ cells seem to be unaffected by the various culture media. These findings indicate that receptors for stimuli of meiotic initiation may exist in germ cells or neighbouring somatic cells. In addition to induction of meiosis, MIS media and forskolin also dose dependently increase the number of male germ cells compared to controls.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418817 TI - Human male infertility may be due to a decrease of the protamine P2 content in sperm chromatin. AB - Basic chromosomal proteins were extracted from the sperm of fertile and infertile human males. The relative proportions of protamine 1, 2, and 3 were determined by scanning microdensitometry following electrophoresis of total protamine in polyacrylamide gels. The findings were as follows: (1) The proportion of protamine P(2 + 3) in sperm obtained from infertile males was lower than that in fertile males. (2) Protamine P(2 + 3) in infertile human males showed reduced affinity to DNA. The possibility that some cases of human male infertility may be due to mutation within the protamine P2 gene is discussed. PMID- 8418818 TI - Pig membrana granulosa cells prevent resumption of meiosis in cattle oocytes. AB - Membrana granulosa was isolated from healthy large antral follicles of prepubertal or cyclic gilts stimulated with PMSG or PMSG and hCG. Ultrastructural observations revealed that pieces of pig membrana granulosa were associated with the basement membrane. The cattle cumulus-enclosed oocytes (COC) were placed in the rolled pieces of the pig membrana granulosa (PMG). After 8 and 24 hr of coculture with PMG from prepubertal gilts, only 16% and 21% of oocytes underwent GVBD, respectively. PMG from PMSG-stimulated cyclic gilts blocked the resumption of meiosis in all COC. The inhibitory effect of heterologous granulosa cells was fully reversible. When COC were initially incubated for 2 and 4 hr, subsequent culture in PMG prevented GVBD in 100% and 36% of oocytes, respectively. This suggests that functional contact between COC and PMG was established during the first 2 hr of coculture. To follow metabolic cooperation between PMG and COC, PMG was prelabeled with 3H-uridine and cocultured with COC. Autoradiography on semithin sections revealed the intensive passage of 3H-uridine from PMG into the cumulus layer and an oocyte. COC placed in PMG after GVBD (8 and 12 hr of an initial incubation) did not extrude the first polar body. PMG isolated from cyclic gilts after PMSG and hCG stimulation also inhibited GVBD of COC. Since nearly all COC placed in PMG isolated 10 and 12 hr after hCG remained in the GV stage after 24 hr of coculture, the hCG stimulation did not substantially diminish the meiosis inhibiting activity of PMG.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418819 TI - Mechanistic studies of the plasma membrane block to polyspermy in mouse eggs. AB - The mechanisms responsible for the plasma membrane associated block to polyspermy in mouse eggs were studied. Reinsemination experiments using zona-free eggs indicated that, after fertilization, the egg plasma membrane is altered such that sperm binding to the egg plasma membrane is blocked, except in the region of the second polar body. Activation of the egg with either ethanol or strontium chloride did not result in a block to polyspermic penetration, as artificially activated eggs displayed identical penetration levels as to nonactivated control eggs. The penetrability of activated eggs was not altered by the presence or absence of the zona pellucida during activation. Lectin staining for egg cortical granule material indicated that activation did cause cortical granule exocytosis; however, activated eggs remained penetrable. These data support the following conclusions: (1) an alteration in the ability of the egg plasma membrane to allow sperm adherence accounts for the block to polyspermy; (2) establishment of the plasma membrane block to polyspermy is sperm dependent, since artificial egg activation does not result in a block response; (3) the contents of the egg's cortical granules do not play a role in the establishment of the plasmalemma block response. PMID- 8418820 TI - Differential chromatin condensation of female and male pronuclei in mouse zygotes. AB - Mouse zygotes or halves of zygotes, containing either a female or a male pronucleus, were fused with ovulated metaphase II oocytes. In 59.7% of the resulting hybrid cells, the pronuclei underwent premature chromosome condensation (PCC). In some of these heterokaryons the 2 pronuclei differed in the dynamics of condensation. Detectability of differential PCC of pronuclei (dPCC) depended on the type of preparation. In hybrids with PCC, produced by fusion of intact zygotes with metaphase II oocytes and processed for whole-mount preparations, one pronucleus was more advanced in the condensation process in 47% of cases. In air dried preparations dPCC was detected in as many as 94% of hybrids. Experiments with the fusion of halves of zygotes with metaphase II oocytes have shown that the differential reaction of pronuclei to condensation factor depended on their parental origin. Maternal chromatin responded faster to the condensation factor and attained more advanced stages of PCC than paternal chromatin. Different responses of the maternal and paternal pronucleus to the condensation factor suggests that the 2 pronuclei are not identical with regard to the organization of chromatin and/or the lamin composition of the nuclear envelope. PMID- 8418821 TI - A major mRNA of the human epididymal principal cells, HE5, encodes the leucocyte differentiation CDw52 antigen peptide backbone. AB - HE5, a very abundant human epididymal gene product, was cloned by a differential screening procedure which employed testis as the primary negative control tissue. Sequencing of the HE5 cDNA showed it to be identical to that encoding the peptide backbone of the human leucocyte differentiation antigen CDw52. Since human genomic DNA contained only one cross-hybridizing fragment, this implies that both products were transcribed from the same gene. Northern blot analysis and in situ transcript hybridization revealed a highly tissue- and cell-type-specific expression pattern for HE5, showing the gene product to originate from epithelial cells of the epididymal and deferent duct. The possible identity of the HE5 product with the CDw52 antigen suggests a link between the immune and reproductive systems. PMID- 8418822 TI - Evidence of new antigens in the mouse cumulus oophorus during preovulatory cumulus expansion. AB - The effects of an antiserum (anti-COC) against ovulated mouse cumulus-oocyte complexes (COC) on in vitro fertilization of mouse oocytes were studied. Preincubation of ovulated COC with various concentrations of anti-COC led to dose dependent impairment of fertilization rates as well as to a decrease in the number of spermatozoa attached to the zona pellucida. Anti-COC was used to probe Western blots of cumulus proteins. These cumuli were obtained from 2 experimental groups of mice corresponding to 2 different maturational stages (preovulatory immature COC or preovulatory mature COC). Two antigens (70 and 80 kDa) present in cumulus intercellular matrix from mature COC were only found as traces in matrix from immature COC. In addition, the protein pattern of the cumulus intercellular matrix was different from that of cumulus cells, whatever the COC maturational stage. These results indicate the appearance of new proteins in the cumulus oophorus during preovulatory expansion and are consistent with the contraceptive action of anti-COC. PMID- 8418823 TI - Functional significance of cumulus expansion in the mouse: roles for the preovulatory synthesis of hyaluronic acid within the cumulus mass. AB - Gonadotropin-stimulated expansion of the mouse cumulus oocyte complex (COC) in vitro, measured with a quantitative videographic method, is comparable to that observed to occur in vivo when medium is supplemented with porcine follicle stimulating hormone (pFSH), 10% fetal bovine serum (FBS), and 2.5 mM glucosamine or optimal concentrations of glutamine and glucose. In the absence of glucosamine, the volumetric expansion of COCs in vitro is never more than 25% of that occurring in its presence. The addition of 6-diazo-5-oxo-1-norleucine (DON), an inhibitor of glucosamine synthesis to medium supplemented with glutamine and glucose, completely inhibits cumulus expansion in vitro. This system was utilized to examine the relationship between cumulus expansion and fertilization rates, and the maintenance of fertilizability in culture. Successful fertilization (as determined by development to the 2-cell stage) was correlated with the quantity and quality of the expanded cumulus mass, and conversely, the spontaneous loss or mechanical removal of the cumulus was correlated with a loss of fertilizability following additional incubation in culture medium. In addition, the i.p. injection of DON inhibited cumulus expansion within the intact follicle and suppressed ovulation. PMID- 8418824 TI - Bovine oocyte development following different oocyte maturation and sperm capacitation procedures. AB - Various procedures have been reported for successful in vitro maturation and in vitro fertilization (IVM/IVF) of bovine follicular oocytes. Direct comparisons of these different recommended procedures have been rare. In this research, involving a total of 5,128 oocytes, a series of experiments were conducted to compare oocyte maturation, fertilization, and development in vitro with 2 maturation systems (with or without added hormones) and 3 types of sperm treatment procedures. Oocytes were collected from ovarian antral follicles (2-7 mm in diameter) within 3 hr after slaughter of cows or heifers. Those with intact or at least 4 layers of cumulus cells were selected for IVM/IVF. Oocytes were incubated for 22 hr in either Medium 199 with 7.5% fetal calf serum (M199 + FCS) alone or M199 + FCS with added hormones (M199 + FCS + H; oFSH 0.5 micrograms/ml, oLH 5.0 micrograms/ml, and E2 1.0 micrograms/ml) at 39 degrees C in 5% CO2 and 95% air. For IVF, frozen-thawed sperm were treated with either 0.1 microM calcium ionophore A23187 (A23187) for 1 min, or 10 or 100 micrograms/ml heparin (H10 or H100) for 15 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418825 TI - Phylogeny of adenylyl cyclases. PMID- 8418826 TI - Roles of second messenger pathways in neuronal plasticity and in learning and memory. Insights gained from Aplysia. PMID- 8418827 TI - The current global situation of the HIV/AIDS pandemic. PMID- 8418828 TI - Expanded programme on immunization. Global Advisory Group--Part II. PMID- 8418829 TI - Acquired immunodeficiency syndrome (AIDS)--data as at 31 December 1992. PMID- 8418830 TI - Effects of exposure to low pH on the lateral mobility of influenza hemagglutinin expressed at the cell surface: correlation between mobility inhibition and inactivation. AB - To investigate the possible role of viral glycoprotein mobility in membrane fusion, fluorescence photobleaching recovery was employed to study the effects of exposure to mildly acidic pH (required to convert many viral fusion proteins to the fusion-active form) on the lateral mobility of influenza hemagglutinin (HA) proteins expressed at the surface of transfected cells. HA proteins from two different strains were compared: X:31 HA, which is activated by a brief exposure to pH 4.9 but is irreversibly inactivated at longer exposure times, and HA from A/Japan/305/57, which is relatively stable to inactivation at this pH [Puri, A., Booy, F.P., Doms, R.W., White, J.M., & Blumenthal, R. (1990) J. Virol. 64, 3824 3832]. The HA proteins from both strains, expressed in CV-1 cells using VS-40 vectors, exhibited relatively unrestricted lateral diffusion at the cell surface. The high mobility persisted following a brief exposure (1 min) to pH 4.9 to mediate conversion to the fusogenic state. Longer times (up to 15 min) of preincubation at pH 4.9 inhibited the lateral mobility of X:31 HA (the lateral diffusion rate was markedly reduced, followed by immobilization) but not of A/Japan HA, whose fusion activity is resistant to such treatment. Inhibition of the lateral mobility of X:31 HA due to preincubation at low pH was not specific to the CV-1 cells and was found also in a CHO cell line stably expressing this protein. The results presented demonstrate a close correlation between loss of mobility and inactivation of fusogenic activity, in accord with the notion that lateral motion of the HA proteins is required for fusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418831 TI - Characterization of purified recombinant fibrinogen: partial phosphorylation of fibrinopeptide A. AB - Human fibrinogen has been expressed in Chinese hamster ovary (CHO) cells using a novel two-step procedure which permits efficient synthesis of engineered variant fibrinogens. CHO cells secreting recombinant fibrinogen were grown in roller bottles and maintained in serum-free media for several months. Recombinant protein was purified from media containing 2-4 micrograms/mL fibrinogen using protamine-Sepharose chromatography. Recombinant fibrinogen was identical to plasma fibrinogen when examined on Coomassie-stained SDS gels run under reducing conditions, and on SDS gels when run under nonreducing conditions after partial or complete plasmin degradation, indicating normal chain assembly, disulfide bond formation, and overall protein conformation. Thrombin digestion of purified fibrinogen led to clot formation with release of normal fibrinopeptides, as identified by HPLC. Fibrinopeptide A released from recombinant fibrinogen was partially phosphorylated (22%), similar to the degree of phosphorylation found for human plasma fibrinogen (20-25%), indicating that partial phosphorylation in inherent in fibrinogen synthesis. PMID- 8418832 TI - Identification of a calcium binding site in the protease domain of human blood coagulation factor VII: evidence for its role in factor VII-tissue factor interaction. AB - Previous studies have identified a putative calcium binding site involving two glutamic acid residues located in the protease domain of coagulation factor IX. Amino acid sequence homology considerations suggest that factor VII (FVII) possesses a similar site involving glutamic acid residues 210 and 220. In the present study, we have constructed site-specific mutants of human factor VII in which Glu-220 has been replaced with either a lysine (E220K FVII) or an alanine (E220A FVII). These mutants were indistinguishable from wild-type factor VII by SDS-PAGE but only possessed 0.1% the coagulant activity of factor VII. Incubation of E220K/E220A FVII with factor Xa resulted in a slower than normal activation rate which eventually yielded a two-chain factor VIIa molecule possessing a coagulant activity of approximately 10% that of wild-type rFVIIa. Amidolytic activity measurements indicated that E220K/E220A FVIIa, unlike wild-type factor VIIa, possessed no measurable amidolytic activity toward the chromogenic substrate S-2288, even at high CaCl2 concentrations. Addition of tissue factor apoprotein, however, induced the amidolytic activity of the mutant molecule to a level 30% of that observed for wild-type factor VIIa. This tissue factor dependent enhancement of E220K/E220A FVIIa amidolytic activity was calcium dependent and required a CaCl2 concentration in excess of 5 mM for maximal rate enhancement. This was in sharp contrast to wild-type factor VIIa which required CaCl2 levels of 0.5 mM for maximal enhancement of tissue factor dependent amidolytic activity. Competition binding experiments suggest that the decrease in amidolytic and coagulant activity observed in the factor VII mutants is a direct result of impaired tissue factor binding.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418833 TI - Amino acid residues controlling acetylcholinesterase and butyrylcholinesterase specificity. AB - Acetyl- and butyrylcholinesterase have 51-54% sequence identity in mammalian species; they exhibit distinct substrate and inhibitor specificities. The crystal structure of acetylcholinesterase enables one to predict folding of related esterases as well as assign residues responsible for differences in substrate specificity. These predictions were tested by expression of esterase chimeras and site-specific mutants using mouse acetylcholinesterase as a template. Chimeras of acetylcholinesterase in which the amino-terminal 174 and the carboxyl-terminal 88 amino acids have been converted to the butyrylcholinesterase sequences still exhibit acetyl-like substrate specificity. Four nonconserved amino acids which are within the central sequence and appear to surround the acyl pocket, F295, R296, F297, and V300, have been mutated alone and in combination to the corresponding residues found in butyrylcholinesterase, L286, S287, I288, and G291. The V300 and R296 mutants slightly enhance butyrylthiocholine hydrolysis while the F295 and F297 mutants, alone and in combination, confer butyrylcholinesterase character by enhancing activity to butyrylthiocholine, and diminishing activity to acetylthiocholine. The F297 mutation eliminates substrate inhibition. F295 and F297 may form a clamp around the acetoxy methyl group. They have distinctive roles in affecting catalysis of the two acylcholines and precisely control acyl ester specificity. Comparison of the susceptibilities of the chimeras and site-specific mutants to cholinesterase-specific inhibitors isoOMPA, ethopropazine, and BW284c51 suggests that inhibitor selectivity for isoOMPA is attributable to residues limiting the size of the acyl pocket, while residues in the amino-terminal domain presumably near the lip of the gorge affect BW284c51 selectivity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418834 TI - Betaine can eliminate the base pair composition dependence of DNA melting. AB - We show that the amino acid analogue betaine shares with small tetraalkylammonium ions [Melchior, W. B., Jr., & von Hippel, P. H. (1973) Proc. Natl. Acad. Sci. U.S.A. 70, 298-302] the ability to reduce or even eliminate the base pair composition dependence of DNA thermal melting transitions. The "isostabilizing" concentration of betaine (at which AT and GC base pairs are equally stable) is approximately 5.2 M. Betaine exerts its isostabilizing effect without appreciably altering the conformation of double-stranded DNA from the B form. The presence of > 5 M betaine also does not greatly change the behavior of DNA as a polyelectrolyte; this lack of effect on electrostatic interactions is expected because betaine exists as a zwitterion near neutral pH. Study of DNA melting transitions in high concentrations of betaine thus allows the experimental separation of compositional and polyelectrolyte effects on DNA melting. As a consequence, betaine solutions can also be used to investigate DNA-protein interactions under isostabilizing (or close to isostabilizing) conditions, which has not been possible using isostabilizing salts. This potential is illustrated by examining the highly salt concentration-dependent interaction of ribonuclease A with DNA in concentrated betaine solutions. PMID- 8418835 TI - Thermal unfolding of a group I ribozyme: the low-temperature transition is primarily disruption of tertiary structure. AB - Little is known about the folding pathways of RNA. A particularly interesting RNA is L-21 Sca I, a linear form of the self-splicing intron from the precursor of the Tetrahymena thermophila large subunit (LSU) rRNA. Thermal unfolding of L-21 Sca I is studied by UV absorption and chemical mapping in 50 mM Na+ and 10 mM free Mg2+ at pH 7.5. UV melting experiments identify two major transitions with maxima at 65 and 73 degrees C. Chemical mapping at the beginning and middle of the first transition suggests it primarily involves disruption of tertiary structure. Phylogenetic comparisons suggest a potential tertiary interaction between loops L2.1 and L9.1a. Chemical mapping and melting experiments on a truncated form of the intron lacking P9.1a, L-21 Nhe I, are consistent with this hypothesis. The results indicate that increasing temperature disrupts tertiary interactions before disrupting secondary structure. This suggests tertiary interactions are weaker than secondary interactions in this case. These results support an important assumption for RNA structure prediction: that secondary structure dominates the free energy of folding. PMID- 8418836 TI - Folding of subtilisin BPN': characterization of a folding intermediate. AB - Subtilisin BPN', an extracellular serine protease from Bacillus amyloliquefaciens, requires a 77 amino acid pro-sequence for correct folding in vivo. We report the observation of a metastable folding intermediate during the refolding of wild-type and a proteolytically inactive mutant subtilisin BPN' that lack the pro-sequence. The addition of the pro-sequence as a separate polypeptide chain results in the folding of the intermediate to the native state. The intermediate state of subtilisin is stable at different temperatures, pH values, and salt concentrations for more than a week and retains its competence for folding. The intermediate state possesses a compactness between that of the native and unfolded states. Although it has native-like secondary structure, it shows no distinct near-UV CD spectrum and has a strongly reduced dispersion in the amide and methyl regions of the 1H NMR spectrum. These indicate considerably less tertiary structure than possessed by the native state. However, the intermediate conformation has regions of stable tertiary structure: it has a high affinity calcium binding site and, after a first noncooperative transition, unfolds with guanidinium chloride in a cooperative process. These results support a folding mechanism for subtilisin BPN' that comprises a high energy transition state, which is lowered by the interaction with the pro-sequence. The similarity to the folding mechanism of alpha-lytic protease supports the hypothesis that a common folding mechanism has been developed through convergent evolution. PMID- 8418837 TI - Comparison of the B-pentamers of heat-labile enterotoxin and verotoxin-1: two structures with remarkable similarity and dissimilarity. AB - We have compared the B-subunit pentamers of Escherichia coli heat-labile enterotoxin (LT) and verotoxin-1 (VT-1). The B-subunits of these bacterial toxins of the AB5 class have virtually no sequence identity and differ considerably in size (69 amino acids in VT-1 versus 103 in LT). They share a number of functional properties: pentamer formation, association with an A-subunit, binding to carbohydrate-containing lipids, and interaction with membranes. The structures of these proteins are very similar in some respects and very different in others. They can be superimposed with an rms deviation of only 1.29 A on the main chain atoms of 52 amino acids (0.98 A on 47 C alpha). Seven out of eight secondary structure elements are retained in the two toxins; only the N-terminal helix of LT is absent in VT-1. A disulfide bridge, which is essential for pentamer formation, is found in both structures, but in slightly different locations. However, the VT-1 B-subunit is much shorter on one side of the toxin, where the proposed membrane binding site of both VT-1 and LT is located. The monomer monomer interface in the pentamer is much larger in LT than in VT-1, making the LT pentamer more stable. The central pores have a different character, and the sugar binding sites are not conserved between the toxins. The evolutionary relationship of the toxins is discussed. PMID- 8418838 TI - Characterization of an independent structural unit in apocytochrome b5. AB - Apocytochrome b5 is a partially folded protein which contains a stable structural unit under native conditions [Moore, C.D., Al-Misky, O.N., & Lecomte, J.T.J. (1990) Biochemistry 30, 8357-8365]. In this work, the fold of the unit was examined by using 1H and 15N-edited two-dimensional NMR spectroscopy. It was found that it contains four of the five beta-strands and two of the six alpha helices present in the holoprotein. The remainder of the structure appears to be mostly unstructured and fluctuating among several conformations. The structural unit is stabilized by a hydrophobic core formed by residues from each of the folded elements of secondary structure. Nuclear Overhauser effects and chemical shift values demonstrated that the unit is structurally similar in the apo- and holoproteins. However, the backbone amide hydrogen exchange was found to be much accelerated in the apoprotein. The paramagnetic relaxation agent HyTEMPO was used to probe the packing of the structure. HyTEMPO has unrestricted access to the empty heme binding site whereas it is unable to penetrate the stabilizing core. It was concluded that addition of the heme is necessary for the last strand to dock properly to the rest of the sheet. The kinetics of refolding of the apoprotein were monitored by stopped-flow fluorescence spectroscopy. Extensive protection of the sole tryptophan residue by docking of the two polypeptide termini occurs in less than 60 ms. It was proposed that apocytochrome b5, with its two-region behavior, might serve as a model for the design of proteins which bind a prosthetic group. PMID- 8418839 TI - Conformational changes in chicken thyroid hormone receptor alpha 1 induced by binding to ligand or to DNA. AB - A classic model of steroid/thyroid hormone receptor activation postulates that a conformational change or "transformation" occurs upon ligand binding as a first step toward regulation of gene transcription. In order to test this model, physical studies have been carried out using purified full-length chicken thyroid hormone receptor alpha 1 (cT3R-alpha 1) expressed in Escherichia coli. Circular dichroism spectroscopic studies reveal that cT3R-alpha 1 adopts a different conformation upon specific binding to a cognate ligand triiodothyroacetic acid as well as to a thyroid hormone response element, an idealized inverted repeat AGGTCA TGACCT. These results suggest that cT3R-alpha 1 may adopt distinct conformations whether free or bound to ligand or to DNA. These states may reflect the changes in the conformation of steroid/thyroid hormone receptors in the signal transduction pathway. PMID- 8418840 TI - Cloning and expression of goldfish opsin sequences. AB - Five opsin cDNA clones were isolated from a goldfish retina cDNA library and sequenced. On the basis of homology to previously characterized visual pigments, one clone was identified as goldfish rod opsin and a second as a goldfish red cone opsin. Two rhodopsin-like clones were found to be similar to the chicken green opsin, a pigment which shares properties with both rod and cone pigments. A fifth clone was equally homologous to human blue cone opsin and human rod opsin. In order to characterize the spectral properties of the encoded pigments, the five clones were expressed in tissue culture cells and the apoproteins reconstituted with 11-cis-retinal. The wavelength of maximal absorption for goldfish rhodopsin is 492 nm and for the fifth pigment, identified as the goldfish blue pigment, 441 nm. Pigments encoded by the two rhodopsin-like clones absorb at 505 and 511 nm and are likely to correspond to the goldfish green pigment previously characterized by microspectrophotometry. The putative red cone opsin cDNA may encode a pigment that is a polymorphic variant of goldfish red since it absorbs maximally at 525 nm. PMID- 8418841 TI - Modification of human hemoglobin with methyl acyl phosphates derived from dicarboxylic acids. Systematic relationships between cross-linked structure and oxygen-binding properties. AB - Human hemoglobin was reacted with five dicarboxylic acid bis(methyl phosphate) reagents under different ligand conditions. The bis(methyl phosphate) reagents tested were derived from fumaric, isophthalic, terephthalic, trans-stilbene-3,3' dicarboxylic, and trans-stilbene-4,4'-dicarboxylic acids. These acyl phosphate mixed anhydrides are anionic electrophiles and will react with N-terminal amino and lysyl epsilon-amino groups to form amides. The major and many of the minor reaction products that result have been isolated and structurally characterized by globin chain and peptide analysis. Products which are not cross-linked, intrachain linked, and interchain singly and doubly cross-linked occur in proportions which depend upon the reaction conditions and reagent. Modifications of the beta chains were limited to the amino groups of beta 1Val, beta 82Lys, and, to a minor extent, beta 144Lys. In the case of the smaller reagents, the amino groups of alpha 1Val, alpha 99Lys, and, to a minor extent, alpha 139Lys were modified. The oxygen binding affinities of most of the major modified hemoglobins have been measured and are characterized by P50 values from about 1/2 to over 5 times that of unmodified human hemoglobin. Most show strong cooperativity with Hill coefficients (n) of 2.0 or greater. Several of the products that are cross-linked between the beta 1Val of one chain and the beta 82Lys of the other chain have oxygen affinities in a physiologically useful range for oxygen transport and delivery. An inverse linear correlation has been found between the log of P50 and bridging distances for the hemoglobins cross-linked between beta 1Val of one chain and the beta 82Lys of the other chain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418842 TI - Characterization of folding intermediates of human carbonic anhydrase II: probing substructure by chemical labeling of SH groups introduced by site-directed mutagenesis. AB - By measurement of UV absorbance, CD spectra, and enzyme activity, we have shown that human carbonic anhydrase II forms a stable and compact folding intermediate at a moderate concentration of guanidine hydrochloride. The major aim of this study was to map the intermediate structure. For that reason, site-directed mutagenesis was used to introduce cysteine residues in various parts of the central beta-structure to give in each case a single cysteine residue. Thereafter, the accessibility of the introduced SH group to specific chemical labeling was used to probe the stability and compactness of the area surrounding each cysteine residue. Our results indicate that the folding intermediate has an ordered native-like secondary structure in the central part of the beta-sheet, whereas the peripheral part of the beta-sheet seems to be less ordered. A large hydrophobic cluster situated between the central beta-sheet core and secondary structure elements on the surface appears to be intact in the intermediate and is remarkably stable even at high GuHCl concentrations (> 5 M). This unusually stable substructure might function as a "seed" during the initiation of the folding process. PMID- 8418843 TI - Differential roles for three conserved histidine residues within the large subunit of carbamoyl phosphate synthetase. AB - Three conserved histidine residues, His-243, His-781, and His-788, located within the large subunit of carbamoyl phosphate synthetase from Escherichia coli were identified by sequence identity comparisons. These three histidine residues were individually mutated to asparagine residues. The H243N mutant enzyme was found to be critical for carbamoyl phosphate synthesis as the mutant protein was unable to synthesize carbamoyl phosphate at a significant rate (< 1/1500). By analysis of the effects of this mutation on the partial reactions catalyzed by CPS, it was determined that this mutation blocked the formation of the carbamate intermediate from carboxyphosphate and ammonia. The H781N mutant enzyme had an order of magnitude reduction for both the rate of carbamoyl phosphate formation and ATP synthesis which is consistent with the proposal that the carboxyl-terminal half of the large subunit is primarily involved in the phosphorylation of the putative carbamate intermediate. This mutation also reduced the effects of the allosteric activator ornithine on the Km parameters for ATP in the overall biosynthetic reaction and ADP in the ATP synthesis reaction. The H788N mutant enzyme is a functional protein which maintains the ability to synthesize carbamoyl phosphate at a rate comparable to that of the wild-type enzyme. The effects of this mutation are 10-fold reductions of the ATP synthetase and the bicarbonate dependent ATPase activities with substantial increases in the Km values for ATP in the full biosynthetic reaction and for ADP in the ATP synthesis reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418844 TI - Conformational and aggregational states of omega-aminoacylmelittin derivatives. AB - Melittin, a 26-residue peptide from bee venom, is known to change from a largely random to a largely alpha-helical conformation as a function of peptide concentration, pH, and ionic strength. In this report, we have determined the effect of displacing the positive charges of the amino groups of N-terminal glycine and lysine residues away from the backbone of melittin in coil-to-helix transitions by using omega-aminoacyl derivatives of melittin. These were prepared by acylating the amino groups of melittin with omega-amino acids to yield the melittin derivatives glycylmelittin (MLT-2), (4-aminobutanoyl)melittin (MLT-4), and (5-aminopentanoyl)melittin (MLT-5), respectively. At pH 7.2, there is a chain length-dependent increase in helicity from MLT to MLT-5. The omega aminoacylmelittin derivatives also show a concentration-dependent increase in helicity at pH 7.2. However, at pH 2.3, a concentration-independent, but chain length-dependent increase in helicity was observed. A hydrophilic derivative glycylglycylmelittin (MLT-GG) and a hydrophobic derivative MLT-5, which have side chains of equal length, show similar helicity, at pH 7.2, but at pH 2.3 MLT-GG shows almost no helicity, while MLT-5 is about 60% helical. The lysyl derivative (MLT-K), which has additional positive charges compared to melittin, behaves much like MLT-2. At pH 7.2, all the derivatives exhibit both cold- and heat-induced denaturation; a significant amount of residual structure is retained in the temperature range 80-100 degrees C. These results are discussed in terms of the electrostatic and hydrophobic interactions involving the side chains. PMID- 8418845 TI - L-29, an endogenous lectin, binds to glycoconjugate ligands with positive cooperativity. AB - The soluble mammalian lactose-binding lectins L-14-I and L-29 are both secreted and bind to oligosaccharides on laminin, a large extracellular matrix glycoprotein containing polylactosamine chains. Because of the potential functional significance of these lectin-laminin interactions, we compared quantitative aspects of L-14-I and L-29 binding to immobilized laminin using recombinant lectins labeled with 125I. We report that the concentration-dependent binding of L-29 exhibits positive cooperativity whereas binding of L-14-I does not. Cooperative binding of L-29 can also occur on glycoconjugate substrates other than laminin and is not dependent on cystine bond formation or aggregation in solution. L-29 contains repetitive sequences within the N-terminal domain not present in L-14-I. This domain is not required for binding activity, but is required for positive cooperativity. Though the precise mechanism of interaction of L-29 with laminin remains to be determined, it apparently results in assembly of a lectin aggregate on the substrate surface, which could have important functional consequences. PMID- 8418846 TI - Biologically active Arg-Gly-Asp oligopeptides assume a type II beta-turn in solution. AB - The sequence Arg-Gly-Asp (RGD) has been found to be the consensus sequence of matrix proteins for binding cell surface receptors (integrins). Studies with synthetic peptides containing the RGD sequence show that the biological activity of these oligopeptides is removed upon a conservative substitution of Glu for Asp in the RGD sequence. Two-dimensional 1H NMR methods were used to investigate the secondary structures in aqueous solution for two such oligopeptides of differing biological activity. The sequence Tyr-Gly-Arg-Gly-Asp-Ser-Pro, which binds to selected integrins, is found to assume a type II beta-turn at both pH 4 and 7. In contrast, the sequence Tyr-Gly-Arg-Gly-Glu-Ser-Pro, which does not interfere with integrin-mediated cell attachment, is found to assume a type I or III beta-turn at both pH 4 and 7. This comparison confirms not only that oligopeptides can assume a secondary structure in aqueous solution, but also that these structures may be important to biological functions. PMID- 8418847 TI - Identification of phosphocholine plasmalogen as a lipid component in mammalian pulmonary surfactant using high-resolution 31P NMR spectroscopy. AB - High-resolution 31P NMR spectroscopy was used to analyze the phospholipid composition of mammalian pulmonary surfactant from two different sources. Under conditions which considerably narrow the usually broad 31P phospholipid signals, solution-phase NMR spectra of these surfactant preparations unequivocally demonstrate that a phosphocholine plasmalogen (i.e., 1-O-(1'-alkenyl)-2-acyl-sn glycero-3-phosphocholine) exists as a major secondary component (approximately 4 mol%) in mammalian pulmonary surfactant. Phosphocholine (PC) plasmalogen was identified in preparations obtained from both adult cow lung surfactant extract as well as in ovine (lamb) fetal pulmonary liquid. PC plasmalogens have not previously been identified any mammalian pulmonary surfactant preparation. The amount of PC plasmalogen in these preparations occurs at fractional levels that are comparable to that of phosphoglycerol (PG), which previously had been thought of as the second-most common phospholipid class in pulmonary surfactant. The presence of PC plasmalogen in pulmonary surfactant may have important physiological ramifications and immediately suggests new directions for biochemical and biophysical investigations of pulmonary surfactant. PMID- 8418848 TI - Cysteine pairing in the glycoprotein IIbIIIa antagonist kistrin using NMR, chemical analysis, and structure calculations. AB - The pairing of the cysteines in disulfide bonds was investigated for the 68 residue RGD-containing protein kistrin, a potent antagonist of the integrin GP IIbIIIa and an inhibitor of platelet aggregation. Kistrin belongs to a family of homologous proteins found in snake venoms termed disintegrins, all of which have a cysteine content. The disulfide pairing of the 12 cysteines was investigated by chemical analysis, NMR spectroscopy, and distance geometry calculations. The data show that the disulfide pairs are 4-19, 6-14, 13-36, 27-33, 32-57, and 45-64. The various means for assigning the disulfide bonds are described, and the results are compared with the cysteine pairings reported for other disintegrin proteins. PMID- 8418849 TI - Effect of glycosphingolipid fatty acid chain length on behavior in unsaturated phosphatidylcholine bilayers: a 2H NMR study. AB - Deuterium-labeled glycosphingolipids, N-lignoceroyl[d47]galactosylceramide (24:0 fatty acid form) and N-stearoyl[d35]galactosylceramide (18:0 fatty acid form) were prepared by partial synthesis. These probe-labeled species, differing only in the lengths of their fatty acids, were compared via 2H NMR with regard to arrangement and behavior in bilayers of the monounsaturated phospholipid 1 stearoyl-2-oleoylphosphatidylcholine (SOPC). Results were used to consider the physical significance of the great range of common acyl chain lengths that is a frequent feature of cell membrane glycosphingolipids. N-lignoceroyl[d47]- and N stearoyl[d35]galactosylceramide were incorporated at concentrations ranging from 5 to 50 mol % into unsonicated phospholipid liposomes, and their spectra were analyzed in the range +73 to -14 degrees C. For the 18:0 fatty acid derivative, first spectral moments, M1, were calculated and plotted as a function of temperature for each sample composition. Spectral inspection for regions of phase coexistence, in conjunction with consideration of M1 curves, permitted derivation of phase diagram boundaries which were then refined using spectral subtraction techniques. The phase diagram for galactosylceramide with short fatty acid in SOPC was compared to the corresponding phase diagram for its long-chain analogue, derived previously in the same fashion [Morrow, M. R., Singh, D., Lu, D., & Grant, C. W. M. (1992) Biochim. Biophys. Acta 1106, 85-93]. The binary phase diagrams referred to above, which reflect the behavior of short- and long-chain glycolipids in a common phospholipid host matrix, displayed important similarities and differences. In fluid membranes, the behavior appeared to be remarkably alike, as reflected in superimposable fluidus curves over the concentration range studied.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418850 TI - 15N- and 13C-labeled media from Anabaena sp. for universal isotopic labeling of bacteriocins: NMR resonance assignments of leucocin A from Leuconostoc gelidum and nisin A from Lactococcus lactis. AB - A procedure for universal 13C and/or 15N labeling of microbial peptides which are produced by fermentation in complex media and its application to two food preserving bacteriocins from lactic acid bacteria are described. Isotopic enrichment of nisin A (from Lactococcus lactis) and of leucocin A (from Leuconostoc gelidum) is readily achieved using a soluble peptone derived from enzymatic hydrolysis (pepsin and chymopapain) of Anabaena sp. ATCC 27899 cells grown on sodium [13C]bicarbonate and/or sodium [15N]nitrate as sole carbon and nitrogen sources. Combustion of this peptone followed by mass spectrometric analysis indicates that 45% of the labeled carbon and 65% of the labeled nitrogen added to the Anabaena culture are utilized in the amino acids of the peptone and that the isotopic purity for both 13C and 15N remains essentially unchanged provided that the cells are grown under argon atmosphere to avoid nitrogen fixation. NMR analyses of [13C,15N]nisin A using H[13C]MQC, H[13C]MBC, 2D INADEQUATE, and H[15N]MQC techniques confirmed 1H spectral assignments previously reported for unlabeled material and readily provided carbon and nitrogen assignments. The results show that universal but not uniform 13C labeling occurs unless the nutrient source is completely isotopically enriched at high level (> or = 98%) because of differential levels of de novo amino acid synthesis. Application of NMR techniques such as TOCSY, DQF-COSY, NOESY, and H[13C]MQC to unlabeled and [13C]leucocin A afforded the complete 1H and 13C assignment. Leucocin A does not possess clearly defined conformational structure in DMSO or aqueous solutions. PMID- 8418851 TI - Two-dimensional 1H NMR studies of 32-base-pair synthetic immobile Holliday junctions: complete assignments of the labile protons and identification of the base-pairing scheme. AB - The labile protons of two 32-base-pair, four-arm models of immobile Holliday junctions have been studied by two-dimensional 1H nuclear magnetic resonance (NMR) spectroscopy. Overlap of resonances in the imino-imino region of two dimensional nuclear Overhauser enhancement (NOE) spectra necessitates the use of a multi-pathway approach for obtaining sequence-specific assignments wherein all possible NOE connectivities to the labile protons are utilized, including those from the 2H of adenine, 5CH3 of thymine, and 5H of cytosine. Resonance assignments are obtained for all slowly exchanging imino and cytosine amino protons. Base-pairing up to and including the junction point is found in all four arms of both Holliday junctions. Several cross-arm NOE connectivities are identified and can be used to infer the geometry of the helical stacking domains. The two Holliday junctions studied, which differ only by the exchange of two base pairs at the branch point, appear to have opposite arm stacking geometries. These assignments form an important part of the critical background for detailed NMR analysis of Holliday junction structure and dynamics. PMID- 8418852 TI - Mapping of the binding interfaces of the proteins of the bacterial phosphotransferase system, HPr and IIAglc. AB - Enzyme IIAglc and HPr are central regulatory and phosphocarrier proteins of the phosphoenolpyruvate:sugar phosphotransferase system (PTS) of bacteria. During phosphoryl transfer from phosphoenolpyruvate to glucose, phosphate is transferred from HPr to enzyme IIAglc. In order to characterize the binding interfaces of the two proteins during phosphate transfer, 15N-edited and 15N-filtered NMR experiments have been recorded for the complex of enzyme IIAglc and HPr from Bacillus subtilis. Uniformly 15N-labeled enzyme IIAglc and nonlabeled HPr were used in these studies. Residues which undergo significant chemical shift changes upon complex formation have been identified for both proteins. The binding interfaces of the two proteins, suggested by the observed chemical shift changes, involve predominantly hydrophobic surfaces near the active site His-15 of HPr and the phosphoryl acceptor His-83 of IIAglc. PMID- 8418853 TI - Rat and human pancreatic islet cells contain a calcium ion independent phospholipase A2 activity selective for hydrolysis of arachidonate which is stimulated by adenosine triphosphate and is specifically localized to islet beta cells. AB - The recent demonstration that myocardial Ca(2+)-independent phospholipase A2 exists as a complex of catalytic and regulatory polypeptides that is modulated by ATP has suggested a novel mechanisms through which alterations in glycolytic flux can be coupled to the generation of eicosanoids which facilitate insulin secretion. To determine the potential relevance of this mechanism, we examined the kinetic characteristics, substrate specificities, and cellular locus of phospholipase A2 activity in pancreatic islets. Rat pancreatic islets contain a Ca(2+)-independent phospholipase A2 activity which is optimal at physiologic pH, preferentially hydrolyzes phospholipid substrates containing a vinyl ether linkage at the sn-1 position, and prefers arachidonic acid compared to oleic acid in the sn-2 position. Rat islet Ca(2+)-independent phospholipase A2 activity is inhibited by the mechanism-based inhibitor (E)-6-(bromomethylene)-3-(1 naphthalenyl)-2H-tetrahydropyran-2-one and is stimulated by ATP. Purification of beta-cells from dispersed pancreatic islet cells by fluorescence-activated cell sorting demonstrated that beta-cells (but not non-beta-cells) contain Ca(2+) independent, ATP-stimulated phospholipase A2 activity. Remarkably, clonal RIN-m5f insulinoma cells, which possess a defect in glucose-induced insulin secretion, contain a Ca(2+)-independent phospholipase A2 which is not modulated by alterations in ATP concentration. Collectively, these results and those of an accompanying paper [Ramanadham et al. (1993) Biochemistry (following paper in this issue)] implicate Ca(2+)-independent phospholipase A2 as a putative glucose sensor which can couple alterations in glycolytic metabolism to the generation of biologically active eicosanoids and thereby facilitate glucose-induced insulin secretion. PMID- 8418854 TI - Inhibition of arachidonate release by secretagogue-stimulated pancreatic islets suppresses both insulin secretion and the rise in beta-cell cytosolic calcium ion concentration. AB - Fuel secretagogues induce hydrolysis of esterified arachidonic acid from pancreatic islet cell phospholipids and accumulation of nonesterified arachidonate at concentrations up to 35 microM. Exogenous arachidonate (5-30 microM) amplifies depolarization-induced insulin secretion from islets. Fuel secretagogue-induced hydrolysis of arachidonate from islet phospholipids occurs in Ca(2+)-free medium, suggesting the possible involvement of a Ca(2+) independent phospholipase. In the companion paper [Gross et al. (1993) Biochemistry (preceding paper in this issue)], we demonstrated that the major islet phospholipase A2 is Ca(2+)-independent, ATP-stimulated, and inhibited by the haloenol lactone suicide substrate (HELSS) (E)-6-(bromomethylene)-3-(1 naphthalenyl)-2H-tetrahydropyran-2-one. Here we demonstrate that HELSS suppressed both release of the arachidonate metabolite prostaglandin E2 and insulin secretion from islets stimulated with D-glucose and the muscarinic agonist carbachol. Both prostaglandin E2 release and insulin secretion were suppressed with similar concentration profiles and time courses. Islet oxidation of [14C] glucose to [14C]CO2, activities of islet lactate dehydrogenase and alanine and aspartate aminotransferases, and carbachol-induced inositol phosphate accumulation in islets were all unaffected by HELSS. Depolarization of isolated beta-cells with 40 mM KCl induced a rise in cytosolic [Ca2+] that was also unaffected by HELSS. In contrast, the 17 mM D-glucose-induced rise in beta-cell [Ca2+] was inhibited by HELSS in a concentration-dependent manner, but that induced by exogenous arachidonate (15 microM) was not. These results suggest that fuel secretagogues activate the islet Ca(2+)-independent phospholipase A2, resulting in release of nonesterified arachidonate, which facilitates Ca2+ entry into beta-cells and promotes insulin secretion. PMID- 8418855 TI - Reactivity of lysyl residues on the (Ca(2+)-Mg2+)-ATPase to 7-amino-4 methylcoumarin-3-acetic acid succinimidyl ester. AB - The (Ca(2+)-Mg2+)-ATPase of sarcoplasmic reticulum was labeled with the succinimidyl ester of 7-amino-4-methylcoumarin-3-acetic (AMCA). Although a large number of residues were labeled, it was found that Lys-492 was labeled preferentially at pH values between 6 and 8, consistent with an unusual environment for this residue. Labeling was reduced in the presence of ATP, suggesting that Lys-492 is in or near the ATP binding site of the ATPase. Other identified residues labeled by AMCA were Lys-35, Lys-135, Lys-218, Lys-371, and Lys-605. It is suggested that these represent surface-exposed lysyl residues. Lys 515, labeled by fluorescein isothiocyanate (FITC), was not labeled by AMCA. Labeling with AMCA at pH 6.0 has no effect on ATPase activity, suggesting that Lys-492 is not essential for activity. The fluorescence of AMCA-labeled ATPase did not change on addition of either ATP in the presence of Ca2+ or Pi in the absence of Ca2+, suggesting that Lys-492 was not affected by any major conformational changes on the ATPase. The efficiency of fluorescence energy transfer between AMCA and FITC labels on the ATPase was unaffected by binding Ca2+ or vanadate, arguing against any large-scale movement of the cytoplasmic domains of the ATPase. PMID- 8418856 TI - Macrophage alpha-actinin is not a calcium-modulated actin-binding protein. AB - alpha-Actinin was purified from rabbit macrophages to apparent homogeneity by a procedure designed to remove other actin-binding proteins. Large bundles of filaments were formed when 1 molecule of alpha-actinin interacted with 10-12 actin monomers. This process involved the successive occupancy of two classes of actin-binding sites with different affinities. The apparent Kd of alpha-actinin for F-actin was unaffected by the addition of 25 microM free Ca2+. Analysis of the influence of increasing Ca2+ concentrations on alpha-actinin-F-actin interactions by low-speed sedimentation assays, low-shear viscosity, and electron microscopy indicated that Ca2+ had a small inhibitory effect in the approximate range of 50-1000 microM. Furthermore, the ability of alpha-actinin to assemble actin filaments into bundles was apparently inhibited only at Ca2+ concentrations which also affected the physical properties of F-actin alone. alpha-Actinin immobilized on a nitrocellulose membrane did not bind detectable amounts of Ca2+. Nevertheless, Ca2+ or Mg2+ binding to alpha-actinin induced small decreases in the fluorescence emission intensity of tryptophan and tyrosine residues. The maximal change induced by Mg2+ was smaller than that observed with Ca2+, but Ca2+ and Mg2+ effects were abolished by the addition of 140 mM KCl. Under near physicological ionic conditions, Ca(2+)-binding sites with an apparent Kd higher than 80-100 microM could not be detected. The results on the functional and physical properties of alpha-actinin are consistent with the hypothesis that Ca2+ decreases alpha-actinin--F-actin interactions by acting both on actin filaments and on cross-linking molecules. Although this conclusion is in contradiction with the generally accepted idea that alpha A is a Ca(2+)-regulated actin-binding protein, it could be predicted from the primary sequence of the two EF-hand-like motifs in the alpha-actinin monomer [Arimura et al. (1988) Eur. J. Biochem. 177, 649-655] based on the crucial role of some Ca(2+)-binding residues as recently demonstrated by point mutations in Ca(2+)-binding sites of calmodulin [Haiech et al. (1991) J. Biol. Chem. 266, 3427-3431]. It is also in agreement with our previous finding that Ca2+ does not affect the behavior of alpha-actinin in actin gel networks from macrophage cytosolic extracts [Pacaud & Harricane (1987) J. Cell Sci. 88, 81-94]. PMID- 8418857 TI - Reaction of the isosteric methylenephosphonate analog of alpha-D-glucose 1 phosphate with phosphoglucomutase. Induced-fit specificity revisited. AB - The phospho form of phosphoglucomutase reacts with the isosteric methylenephosphonate analog of alpha-D-glucose 1-phosphate to produce the corresponding analog of alpha-D-glucose 1,6-bisphosphate plus the dephosphoenzyme. In a coupled reaction, kcat/Km = 1.7 x 10(3) M-1 s-1, which is about 2 x 10(-5) times that for the corresponding reaction with alpha-D-glucose 1 phosphate. The decrease in kcat/Km is divided more or less evenly between less efficient PO3- transfer and decreased binding, although smaller phosphates and phosphonates bind approximately equally. There is a much smaller difference in the binding of glucose 1-methylenephosphonate 6-phosphate and glucose 1,6 bisphosphate to the dephosphoenzyme: the binding ratio is < 1:35 when the glucose ring is oriented similarly. Preferred binding patterns for a number of substrates/inhibitors, studied by 31P NMR and UV-difference spectroscopy, suggest that in the ground state the phosphonate group is tolerated to a much greater extent at the catalytic subsite than at the phosphate-binding subsite, where binding specificity appears to be directed toward a tetrahedral-PO3(2-) group attached to a bridging atom that can act as a hydrogen-bond acceptor. Binding specificity at the catalytic subsite apparently is directed toward a different array, possibly (-O...PO3...O-)2-. Some of these results are considered in terms of a modified version of the "induced fit" concept of enzymic specificity, which is reexamined in view of implied thermodynamic restrictions. The internal rearrangement whereby the positions of the anionic groups of the phosphate/phosphonate are exchanged is compared with the analogous rearrangements involving glucose 1,6-bisphosphate and 1,4-butanediol bisphosphate. The supplementary material describes a three-step synthesis of 1-deoxy-alpha-D glucose 1-methylenephosphonate together with a procedure for phosphorylating the phosphonate to produce an analog of alpha-D-glucose 1,6-bisphosphate and also describes a facile procedure for the qualitative conversion of organic phosphonates to inorganic phosphate. PMID- 8418858 TI - Z-DNA structure of a modified DNA hexamer at 1.4-A resolution: aminohexyl-5' d(pCpGp[br5C]pGpCpG). AB - Oligonucleotides with modification at the 5'-end have been used for various biochemical applications. As a first step to better assess the effects of those modifications on DNA conformation, we determined at 1.4-A resolution the left handed Z-DNA structure of a DNA hexamer, aminohexyl-5'-d(pCpGp[br5C]pGpCpG), by X ray diffraction analysis. This hexamer was crystallized in the monoclinic C2 (a = 51.13 A, b = 18.44 A, c = 34.67 A, and beta = 120.9 degrees) space group. Its structure has been refined by the restrained least-squares refinement to a final R factor of 0.164 using 3727 [> 2.0 sigma (F)] observed reflections. The overall conformation of the double helix resembles that of the canonical Z-DNA. The terminal 5'-phosphate groups of the dC residues adopt conformations (beta approximately 180 degrees and gamma approximately 60 degrees) similar to phosphodiester's conformation of the internal dC residues. Two types of interhelical stackings are observed, one of which may serve as a model for a single-strand nick in the backbone of DNA double helix. A barium ion is found to bridge two side-by-side Z-DNA helices by coordinating to the O6 and N7 atoms of two guanines simultaneously. This "cross-linking" ability of barium ion may be a useful property in promoting the reversible aggregation of nucleic acids. PMID- 8418860 TI - Chemical and enzymatic oxidation of benzimidazoline-2-thiones: a dichotomy in the mechanism of peroxidase inhibition. AB - Derivatives of imidazole-2-thiones block reactions catalyzed by thyroid peroxidase (TPX) and the closely related lactoperoxidase (LPX), and this property is used therapeutically to treat hyperthyroidism. The reactions of a series of benzimidazoline-2-thiones with chemical and enzymatic oxidants were investigated to probe systematically the mechanism of inhibition. Oxidation of benzimidazoline 2-thione (I) and 1-methylbenzimidazoline-2-thione (II) with 3-chloroperbenzoic acid (PBA) yielded reaction products and stoichiometry consistent with benzimidazole-2-sulfenic acids as reactive intermediates. The N,N'-disubstituted nature of 1,3-dimethylbenzimidazoline-2-thione (III) precludes sulfenic acid formation by tautomerization, and the oxidation of III with PBA yielded products and stoichiometry that were consistent with a benzimidazole-2-sulfonyl ylide as the reactive intermediate. I and II are suicide inhibitors of LPX and TPX, but III was found to inhibit only peroxidase-catalyzed iodination reactions by an alternate substrate mechanism. These results provide support for the hypothesis that imidazole-2-sulfenic acids are important reactive intermediates in the suicide inactivation of TPX and LPX and relate the chemical reactivity of the inhibitor with both the potency and mechanism of inhibition. These results suggest that 1,3-disubstituted thiourea derivatives represent a new class of potential antihyperthyroid drugs that block TPX-catalyzed tyrosine iodination but do not cause irreversible enzyme inactivation. PMID- 8418859 TI - Structural changes at the metal ion binding site during the phosphoglucomutase reaction. AB - An electron density map of the reactive, Cd2+ form of crystalline phosphoglucomutase from X-ray diffraction studies shows that the enzymic phosphate donates a nonbridging oxygen to the ligand sphere of the bound metal ion, which appears to be tetracoordinate. 31P and 113Cd NMR spectroscopy are used to assess changes in the properties of bound Cd2+ produced by substrate/product and by substrate/product analog inhibitors. The approximately 50 ppm downfield shift of the 113Cd resonance on formation of the complex of dephosphoenzyme and glucose 1,6-bisphosphate is associated with the initial sugar-phosphate binding step and likely involves a change in the geometry of the coordinating ligands. This interpretation is supported by spectral studies involving various complexes of the active Co2+ and Ni(2+)-enzyme. In addition, there is a loss of the 31P 113Cd J coupling that characterizes the monophosphate complexes of the Cd2+ enzyme either during or immediately after the PO3- transfer step that produces the bisphosphate complex, indicating a further change at the metal binding site. The implications of these observations with respect to the PO3- transfer process in the phosphoglucomutase reaction are considered. The apparent plasticity of the ligand sphere of the active site metal ion in this system may allow a single metal ion to act as a chaperone for a nonbridging oxygen during PO3- transfer or to allow a change in metal ion coordination during catalysis. A general NMR line shape/chemical-exchange analysis for evaluating binding in protein-ligand systems when exchange is intermediate to fast on the NMR time scale is described. Its application to the present system involves multiple exchange sites that depend on a single binding rate, thereby adding further constraints to the analysis. PMID- 8418861 TI - Mechanism of activation of protein kinase C: roles of diolein and phosphatidylserine. AB - We studied the roles of lipid concentration, phosphatidylserine (PS), and diolein (DG) contents, as well as Ca2+ concentration, on the partitioning of protein kinase C (PKC) between aqueous and membrane environments as well as the relationship of this partitioning to the activation of the enzyme. Physiological concentrations of 1 mol % DG increased the apparent binding constant of PKC to the 3:1 PC/PS membrane 500 times. This increase was proportional to the mol % DG. Over 50% PKC was bound to that membrane at micromolar concentrations of Ca2+ and physiologically relevant total concentration of lipid only when 1 mol % DG was included. PKC bound either to PS alone or to PS and DG was enzymatically competent; however, the rate of phosphorylation was doubled in the presence of 1 mol % diolein. The dependence of PKC binding on the mol % PS was highly sigmoidal. The Hill coefficient was in the range of 4-6, with the higher values found at the lower lipid concentrations. These results suggest that the observed apparent cooperativity is due, at least in part, to the change in dimensionality when PKC binds to the membrane. PMID- 8418862 TI - Use of an antisense oligonucleotide to inhibit expression of a mutated human procollagen gene (COL1A1) in transfected mouse 3T3 cells. AB - A series of antisense oligonucleotides were developed to inhibit specifically expression of a mutated exogenous gene for collagen without inhibiting expression of an endogenous gene for the same protein. The test system consisted of mouse NIH 3T3 cells that were stably transfected with an internally deleted construct of the human gene for the pro alpha 1(I) chain of type I procollagen [Olsen et al. (1991) J. Biol. Chem. 266, 1117]. The target site was a region at the 3' end of exon 1 and the first few nucleotides of intron 1 of the exogenous human gene that differed in sequence by nine nucleotides from the sequence of the endogenous mouse gene. Expression of the two genes was assayed by Western blot with cross reacting antibodies and by steady-state levels of mRNAs. None of the oligonucleotides were effective in concentrations up to 25 microM when administered without any carrier. However, when administered with 5 or 10 micrograms/mL lipofectin, one of the oligonucleotides in concentrations of 0.1 0.2 microM inhibited expression of the exogenous gene from 50% to 80% without significant inhibition of expression of the endogenous gene. Also, a missense version of the same oligonucleotide had no significant effect, and the inhibition observed with the most effective oligonucleotide was abolished by a single base change. Time course experiments indicated that, after a 4-h treatment, inhibition appeared at 8 h and persisted for at least 22 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418863 TI - Structural characterization of the gene encoding rat 25-hydroxyvitamin D3 24 hydroxylase. AB - The structural gene encoding 25-hydroxyvitamin D3 24-hydroxylase (P-450cc24) was isolated from the rat genomic DNA. It spans approximately 15 kb, is composed of 12 exons, and was demonstrated by Southern blot analysis to be present as a single copy. One major T residue was identified at the cap site, a putative TATA (ATAAATA) box was located at position -30, and a putative CCAAT box was at -58. Four possible vitamin D responsive elements that may be involved in regulation of 24-hydroxylase expression were found in the 5'-flanking region. Alignment with mitochondrial P-450 proteins showed that 7 out of 11 intron insertion sites of P 450cc24 gene occupied positions identical with those in the CYP11 family (P 450scc, P-450(11 beta)). The structure of the gene is discussed in relation to present knowledge about the mechanism of regulation of the 25-hydroxyvitamin D3 24-hydroxylase and calcium homeostasis. PMID- 8418864 TI - Membrane translocation of diphtheria toxin A-fragment: role of carboxy-terminal region. AB - The C-terminal end of diphtheria toxin A-fragment was altered and the consequences for toxicity and translocation of the A-fragment to the cytosol were studied. Mutations and deletions in the protease-sensitive, disulfide-bridged region linking the two functional parts of the toxin, the A- and B-fragments, reduced the toxicity of the protein as such, but when the mutant toxins were cleaved ("nicked") by trypsin before being added to cells, the toxicity was restored. Prevention of disulfide formation by removal of Cys186 resulted in complete loss of toxicity. To circumvent the nicking step, toxin was formed by reconstitution from separate A- and B-fragments where the A-fragments varied in the C-terminal sequences. The amino acids C-terminal to Cys186 were found not to be required for translocation. Furthermore, both charged and uncharged residues near the C-terminal end were compatible with translocation. The data indicate that the C-terminal amino acid sequence is not decisive for translocation of diphtheria toxin A-fragment to the cytosol. PMID- 8418865 TI - Anthropometric and anthroposcopic findings of the nasal and facial region in cleft patients before and after primary lip and palate repair. AB - Two populations were studied and compared to determine the effects of cleft lip and palate surgery on the nose and face. In the first, three anthropometric measurements, two nasal and one facial, were taken before primary lip repair from infants with complete unilateral cleft lip and palate (UCLP) and with bilateral cleft lip and palate (BCLP). In the second, ten measurements were taken from the nose and face of patients surgically treated for UCLP and BCLP. The high frequency of noses disproportionately wide in relation to their height in both cleft types before primary lip repair greatly decreased after surgery. Among the residual deformities after surgery for UCLP, nostril floor width asymmetry was the most frequent, followed by columella length asymmetry, flat nasal bridge, wide soft nose, flat nasal tip, and small nasal tip protrusion. In the BCLP patients, nostril floor width asymmetry was also the most common stigma, followed by flat nasal tip, wide soft nose, columella length asymmetry, flat nasal bridge and bilaterally angled alae, and small nasal tip protrusion. A subnormally flat upper face inclination was observed in UCLP and BCLP patients. Quantitative determination of these nasal stigmata in cleft lip and palate patients who have undergone primary lip repair provides valuable information for surgical correction of the cleft soft-tissue deformities. PMID- 8418866 TI - Awareness of the cleft palate team. PMID- 8418867 TI - Fetal cleft lip repair in rabbits: long-term clinical and cephalometric results. AB - We have developed a model for fetal cleft lip (CL) repair in rabbits. To date, the in utero CL procedure has been performed on 174 fetuses in 98 pregnant does. Details of the model, wound healing characteristics, and early growth results have been published previously. In this study, we report long-term clinical and cephalometric findings in 23 fetuses who underwent the fetal CL procedure, were born alive, and survived until completion of growth. The surgically created and repaired CL in fetal rabbits described here resulted in healing without scar formation. The deformity varies from an incomplete to a severe complete cleft, resembling the clinical spectrum of spontaneous clefts in humans. Cephalometric studies indicate that there were no statistically significant differences in premaxillary width, anterior maxillary length, or anterior and posterior maxillary width among control, unrepaired, and repaired animals. Documentation of this phenomenon in higher animals is necessary before the technique can be applied to humans with cleft lip. PMID- 8418868 TI - A six-center international study of treatment outcome in patients with clefts of the lip and palate: evaluation of maxillary asymmetry. AB - This investigation was carried out by the European Cleft Lip and Palate Research Group. The purpose of this part of the investigation was to compare and evaluate maxillary asymmetry in children born with complete unilateral cleft lip and palate treated at cleft palate centers with different surgical management. Posteroanterior radiographs from three of the six participating centers were included in the investigation. Statistically significant differences were found in the symmetry of the anterior part of the maxillary complex. Children with a primary bonegrafting procedure involved in their primary treatment procedure had a more symmetric dentoalveolar development. Children from centers with primary surgical procedures including a vomer plasty and no involvement of the alveolar process had a more asymmetric development with a tilted premaxilla and a deviating inclination of the central incisors. PMID- 8418869 TI - Morphologic effect of preoperative maxillofacial orthopedics (T-traction) on the maxilla in unilateral cleft lip and palate patients. AB - Sixty-eight children born with complete unilateral cleft lip and palate were studied using dental casts taken at ages 0-0.1; 0.2-0.4; and 0.5-0.6 years of age. They were all treated with preoperative maxillofacial orthopedics using an external device (T-traction). The treatment was started after the first model was taken. Dental casts were analyzed regarding the morphology of the cleft region and the maxillary segments before and after treatment. The measurements were compared with measurements on dental casts of nontreated cleft children of similar age. The results suggest that a more normal anatomy of the cleft region occurs during the first 6 months of life whether preoperative maxillofacial orthopedics (T-traction) is used or not. However, this normalized growth seems to occur faster with the T-traction. PMID- 8418870 TI - Results of a survey of cleft palate teams concerning the use of nasendoscopy. AB - A national survey was conducted concerning methods used for the evaluation of velopharyngeal function with emphasis on the role of nasendoscopy. Forty-five percent of questionnaires were returned. Ninety percent of the responding teams indicated that nasendoscopy was available. Sixty-one percent agreed that endoscopy was an important clinical tool and not solely a research tool. The majority (59%) considered 3 to 5 years of age to be the youngest, appropriate age for referral. Ninety percent agreed that nasendoscopy was indicated for difficult diagnostic problems and 41% reported endoscopic studies were appropriate for all patients for whom secondary palatal management is planned. The results of this survey suggest that endoscopic assessment of velopharyngeal function is used routinely as an adjunct to the perceptual evaluation of speech and has become the standard of care among cleft palate teams for difficult diagnostic cases. However, the data also indicate that increased availability does not necessarily assure optimal use. PMID- 8418871 TI - Survey of speech-language pathologists' training, experience, and opinions on nasopharyngoscopy. AB - A questionnaire on nasopharyngoscopy was completed by 93 speech-language pathologists randomly selected from the Directory of the American Cleft Palate Craniofacial Association. Although the majority rated nasopharyngoscopy as important in the assessment of velopharyngeal function, and believed that it should be performed by speech-language pathologists, the majority do not perform nasopharyngoscopic examinations. Moreover, 40% had no academic preparation and 20% had no clinical experience in nasopharyngoscopy. Implications of these findings concerning exposure to nasopharyngoscopy in speech-language pathology graduate training programs and in continuing education programs for practicing speech-language pathologists are discussed. PMID- 8418872 TI - Correction of velopharyngeal insufficiency by pharyngeal augmentation using autologous cartilage: a preliminary report. AB - Twenty patients with velopharyngeal insufficiency (VPI) were treated with autologous costal bone or cartilage implants. Videofluoroscopy and videonasopharyngoscopy studies were used to identify candidates for the procedure. The specific size and shape of the gap as well as an appropriate location for the implant were determined in those patients for whom videonasopharyngoscopy was used. A piece of costal bone or cartilage was implanted into a preselected site in the posterior pharynx. Speech and voice evaluation were conducted preoperatively and 8 weeks postoperatively. Hypernasality and audible nasal emissions were completely eliminated in five patients. Four patients had no postoperative change in speech quality. In the remaining patients, improvement occurred without elimination of VPI. The use of costal bone was discontinued after the ninth patient. One infection occurred with complete resorption of the costal bone graft. Resorption has not been a problem with costal cartilage. No extrusion has occurred, and there have been no infections. Overall speech improvement in this group of children (80%) compared favorably with other reports on pharyngeal augmentation. Costal cartilage appears to be a superior choice for implant material. PMID- 8418874 TI - Phonologic processes in children with cleft palate. AB - This study examined the phonologic process usage of 3-, 4- and 5-year-old children with cleft palate. Sixty children served as subjects: 30 children with cleft palate (with or without cleft lip) and 30 noncleft palate children. The children's whole word productions were analyzed for frequency and type of phonologic process usage. Results indicated that the 3- and 4-year old children with cleft palate exhibited more instances of process usage, compared to their noncleft peers. The 5-year-old cleft and noncleft groups were similar in total instances of process usage. Further, the children with cleft palate employed common phonologic processes; however, some processes were noted more frequently in the speech of the 3-year-old children with cleft palate. PMID- 8418873 TI - Long-term speech results of cleft palate patients with primary palatoplasty. AB - This investigation examined the influence of cleft type, type of surgery, age at surgery, and gender on speech proficiency of 204 patients with cleft palate who required only primary palatoplasty. Speech measures were obtained for each subject from at least three annual examinations between the ages of 4 and 16 years. Neither age at surgery nor type of surgery were discriminating factors. The less extensive cleft type, i.e., soft palate only, was associated with greater rates of change in the performance variables than were the other three cleft types. Females showed greater rate changes than males. PMID- 8418875 TI - Speech in unilateral and bilateral cleft palate patients from Stockholm. AB - The speech of 84 patients with complete unilateral cleft lip and palate and 19 patients with complete bilateral cleft lip and palate was judged by professional listeners and compared with a control group of 40 noncleft subjects. The unilateral cleft group consisted of two subgroups: one group of 45 patients, who were treated with presurgical orthopedics before primary surgery, and one group of 39 patients, who were not. The speech of the patients and the noncleft subjects was tape recorded and randomly mixed prior to listener judgments. No significant differences in articulation or resonance were found between the subgroups of unilateral cleft patients. The results also indicated that the bilateral cleft patients had poorer speech and needed more speech therapy than the unilateral cleft patients. All cleft patients were found to have poorer speech than the noncleft subjects in spite of considerable speech therapy and complementary surgical treatment. This has resulted in a change in the Stockholm approach toward earlier palatal surgery, tailor-made pharyngeal flap operations, and earlier parental information and treatment of articulatory deviations. PMID- 8418876 TI - Component approach for partitioning nasal airway resistance: pharyngeal flap case studies. AB - Individuals with craniofacial anomalies often have nasal cavity and/or velopharyngeal constriction. The purpose of this clinical report was to illustrate a technique for partitioning nasal airway resistance into its nasal cavity and velopharyngeal components. This information would be helpful in determining intervention to reduce high nasal airway resistance as well as in providing information about the outcome of corrective procedures to establish velopharyngeal competence for speech. Data from two pharyngeal flap patients seen before and after surgery were utilized in this illustration. These case studies illustrate the usefulness of component resistance measures in quantifying nasal airway patency before and after corrective surgery for velopharyngeal function. PMID- 8418877 TI - Combined effects of severity of cleft impairment and facial attractiveness on social perception: an experimental study. AB - Children and adolescents provided their impressions of stimulus faces that systematically varied in attractiveness and severity of cleft impairment. The results indicated that facial attractiveness is a consistent characteristic of cleft-impaired faces. However, facial attractiveness did not moderate the negative impact of cleft impairment on social perception. Reduced severity of impairment did moderate the negativity of social perception. The implications of these findings for our understanding of the effects of cleft impairment on social perceptions are discussed. PMID- 8418878 TI - Parents' attitudes toward family involvement in cleft palate treatment. AB - Families exert a powerful influence upon their members and can be a significant resource in cleft palate treatment. The purposes of this study were to gather information about (1) the characteristics and attitudes of families whose children are treated for cleft palate; (2) the extent to which parents participate in decisions about treatment; and (3) parents' ideas about their participation. A 32-item survey questionnaire was distributed to 75 parents in Florida, Louisiana, Minnesota, and Pennsylvania. Forty-two survey forms (56%) were completed and returned for analysis. The majority of parents (57%) were satisfied with their present involvement in team treatment services; however, 36% wished for more involvement, and 7% wished for less involvement. PMID- 8418879 TI - Evaluation of maxillary dental arch form in unilateral clefts of lip, alveolus, and palate from one month to four years. AB - This study evaluated the early changes of maxillary alveolar arches of operated unilateral cleft lip and palate patients. Dental casts were available at four age increments. Triangular flap cheiloplasty was carried out at an early age. Two stage palatoplasty by vomer flap and soft palate closure took place later. Prior to lip repair, the alveolar arches were classified according to the relationship between greater and lesser segments. Almost a quarter had overlap of the alveolar segments with no contact between the alveolar ridges at the cleft site; some had no overlap with contact of the alveolar segments in the cleft region; almost a quarter had both overlap of the alveolar segments and contact; and almost half had no overlap of the segments and the alveolar ridges were not in contact at the cleft site. After lip repair, the arch relationships were examined and the percentage of patients in each of the four groups indicated a moulding effect of lip repair on the alveolar segments. This moulding effect caused the alveolar segments in most patients to be in contact at the cleft site. Most of these also had segment overlap. All patients were re-examined shortly after palatal repair. The trend for segment overlap and contact continued after palate surgery. However, when all patients were seen at age 4, percentages of patients in each group indicated that previous overlap of segments improved to a more desirable nonoverlapped relationship in approximately half of the patients. The other half continued to demonstrate arch collapse, in excess of what would be considered ideal ridge relationship. PMID- 8418880 TI - Median cleft of the lip: its significance and surgical repair. AB - Median cleft lip is a midline vertical cleft through the upper lip in the absence of a prolabial remnant. This may occur as a sporadic event or be part of an inherited sequence of anomalies. A failure of formation or fusion of the medial nasal prominences derived from the frontonasal prominence is ultimately responsible for this aberration. Two categories of dysplasia are associated: (1) frontonasal deformity associated with hypotelorism and (2) median facial cleft syndrome associated with hypertelorism. A patient presents with median cleft lip, mild bifid nose, and hypertelorism. Following surgical reconstruction, a good result is achieved. The embryology, implications for associated abnormalities, and surgical technique for treating these cases are discussed. PMID- 8418881 TI - Ear malformation and hearing loss in patients with Treacher Collins syndrome. AB - Although the hearing loss of patients with Treacher Collins syndrome is well documented, few studies have reported jointly on their hearing loss and ear pathology. This paper reports on the hearing loss and computerized tomography (CT) assessments of ear malformations in a large pediatric series of patients with Treacher Collins. Of the 29 subjects assessed by the Craniofacial Program between 1986 and 1990, paired audiologic and complete CT assessments were available for 23 subjects. The external ear canal abnormalities were largely symmetric, either bilaterally stenotic or atretic. In most cases, the middle ear cavity was bilaterally hypoplastic and dysmorphic, and ossicles were symmetrically dysmorphic or missing. Inner ear structures were normal in all patients. The majority of patients had a unilateral or bilateral moderate or greater degree of hearing loss and almost half had an asymmetric hearing loss. The hearing loss of all subjects was conductive, except for three whose loss was bilateral mixed. Two types of bilaterally symmetric hearing loss configurations, flat and reverse sloping, were noted. Conductive hearing loss in patients with Treacher Collins is mainly attributable to their middle ear malformations, which are similar for those of patients with malformed or missing ossicles. PMID- 8418882 TI - The effect of compounds associated with cigarette smoking on the secretion of lipoprotein lipid by HepG2 cells. AB - Cigarette smoking is associated with significant alterations in serum levels of lipids and lipoproteins, however the biochemical mechanisms responsible for this effect are poorly understood. One possibility is that compounds which are present (nicotine) or elevated (free fatty acid, epinephrine, cortisol) in the blood of smokers might contribute to the observed effects by modulating hepatic lipoprotein secretion. The human hepatoma cell line HepG2 was used as a model system to investigate this question. Pre-incubation of the cells for 24 h with 1 mM oleate caused increases in VLDL cholesterol secretion (0.27 to 0.37 micrograms/mg cell protein/24 h, P < 0.01) and decreases in LDL and HDL cholesterol secretion (1.0 to 0.7 micrograms/mg cell protein/24 h, P < 0.02 and 4.1 to 2.5 micrograms/mg cell protein/24 h, P < 0.02, respectively). Incubation with 1.0 microM dexamethasone caused an increase in HDL cholesterol secretion (2.46 to 3.83 micrograms/mg cell protein/24 h, P < 0.05), whereas incubation with 1.0 microM epinephrine caused an increase in LDL cholesterol secretion (0.9 to 1.8 micrograms/mg cell protein/24 h, P < 0.01). Incubation with either dexamethasone or epinephrine caused significant increases in total media cholesterol (P < 0.02), whereas preincubation with oleate did not. Nicotine (10 microM) did not affect lipoprotein lipid secretion. In conclusion, the effects of oleate and epinephrine on the lipoprotein lipid levels secreted by HepG2 cells were consistent with the altered serum lipoprotein levels observed in smokers, while the effects of dexamethasone, a cortisol analogue, were not. PMID- 8418883 TI - Elevated N-myristoyl transferase activity is reversed by sodium orthovanadate in streptozotocin-induced diabetic rat. AB - N-Myristoyl transferase (NMT) is the enzyme that covalently modifies several proteins important in signal transduction. Streptozotocin-induced diabetes resulted in a 2-fold increase in NMT activity from rat liver as compared to control animals. Administration of sodium orthovanadate to the diabetic rats reduced the activity of the NMT to 75-120% of the control values. Elevated NMT activity was observed with both cAMP-dependent protein kinase-derived and pp60src derived peptide substrates. No significant change in the apparent Km was observed with the cAMP-dependent protein kinase-derived peptide substrate. Unlike in rat brain, in all conditions highest NMT activity was observed in the particulate fraction of rat liver. PMID- 8418884 TI - Tissue-specific changes in lipid composition and lipoprotein lipase activity during the development of the chick embryo. AB - Lipoprotein lipase was present at a high specific activity in adipose tissue and heart of the chick embryo at the 14th day of development. The enzyme was also present in skeletal muscle but was absent from brain and liver. Major increases in the activity of lipoprotein lipase in adipose tissue and heart occurred from day 12 of development, concomitant with the beginning of the period of lipid uptake from the yolk. These results suggest that lipoprotein lipase may be involved in the utilisation of yolk-derived lipid by the tissues of the embryo. Relatively high levels of docosahexaenoic acid (22:6(n--3)) were present in the triacylglycerol isolated from plasma, adipose tissue, heart and liver. The relative proportions of this fatty acid in the triacylglycerol of the different tissues may be explicable in terms of the substrate specificity of lipoprotein lipase. PMID- 8418885 TI - Determination of lipid hydroperoxides in native low-density lipoprotein by a chemiluminescent flow-injection assay. AB - Lipid hydroperoxides have been implicated in the pathogenesis of atherosclerosis. This work was therefore set up to obtain a fast and specific chemiluminescent assay for measuring hydroperoxides in native low-density lipoprotein (LDL). The apparatus was a complete HPLC system including two pumps, an autosampler, a computer and a chemiluminescent detector with a T-mixing coil in the place of the column. Samples were injected from the autosampler and mixed with luminescent reagent (3 microM luminol and 1 microM microperoxidase in 0.1 M carbonate buffer (pH 10)) in the T-piece. To generate a calibration curve, linoleic acid hydroperoxide was obtained by incubating soybean lipoxygenase with linoleic acid. The calculated conjugated diene concentration was in good agreement with the nominal linoleic acid hydroperoxide concentration. The chemiluminescence was linear with the amount of linoleic acid hydroperoxide injected and the detection limit was about 3 pmol linoleic acid hydroperoxide. The chemiluminescence induced by copper-oxidized LDL was linear with concentration; the detection limit, when compared with linoleic acid hydroperoxide, was similar. The reproducibility of the linoleic acid hydroperoxide and of oxidized LDL hydroperoxide was examined in single pools. The coefficient of variation on the triplicates of each pool was about 3%. The titre of the linoleic acid hydroperoxide and oxidized LDL peroxides was quite stable for at least 10 days when stored under argon at 4 degrees C in the presence of EDTA. The mean value of the LDL hydroperoxides in 16 control subjects was 145.20 +/- 98.81 pmol/mg LDL protein. In conclusion, the microperoxidase-luminol-dependent chemiluminescence flow-injection assay is a rapid, sensitive and selective method for measuring lipid hydroperoxides in native LDL. PMID- 8418886 TI - Isolation of pigment-free bulk lipids from thylakoids. AB - Lipids from spinach thylakoids were extracted with chloroform/methanol and separated from pigments in a single chromatographic step run at 5 degrees C using silicic acid adjusted to pH 8. The isolated lipid fraction contained essentially the same amounts of individual lipids as in the initial extract. It contained less than 0.1% of the initial chlorophylls and carotenoids. PMID- 8418887 TI - An enzymatic explanation of the differential effects of oleate and gemfibrozil on cultured hepatocyte triacylglycerol and phosphatidylcholine biosynthesis and secretion. AB - Incubation (1-4 h) of primary cultures of adult rat hepatocytes with gemfibrozil (0.1-1.0 mM) significantly decreased the: (1) incorporation of [1,3-14C]glycerol into cellular triacylglycerol (30%); (2) secretion of labeled (VLDL) triacylglycerol (4-fold); and (3) oleate-induced rise in triacylglycerol biosynthesis and secretion. Gemfibrozil also increased the: (1) incorporation of labeled glycerol into cellular phosphatidylcholine (2-fold); and (2) secretion of labeled (HDL) phosphatidylcholine (10-fold). The gemfibrozil-dependent increase in the flux of labeled diacylglycerol into phosphatidylcholine is rapid (15 min) and associated with a 2-fold increase in membrane-bound phosphocholine cytidylyltransferase activity. A phosphocholine cytidylyltransferase-mediated rise in cellular CDP choline content may explain the gemfibrozil-dependent rise in phosphatidylcholine biosynthesis since homogenates of monolayers incubated with CDP choline preferentially incorporate labeled diacylglycerol into phosphatidylcholine rather than triacylglycerol. Therefore, the triacylglycerol lowering potential of gemfibrozil may be due in part to its ability to shunt liver cell diacylglycerol into phosphatidylcholine rather than triacylglycerol. These results suggest that CDP choline may be a key regulator of the diacylglycerol branchpoint, since diacylglycerol is primarily incorporated into phosphatidylcholine or triacylglycerol depending on whether CDP choline is or is not available. PMID- 8418888 TI - Hepatic monoacylglycerol acyltransferase activity in HA1 and HA7 hepatoma/hepatocyte hybrid cells: regulation by insulin and dexamethasone and by cell density. AB - Hepatic monoacylglycerol acyltransferase (MGAT) (EC 2.3.1.22) is a developmentally-expressed enzyme that catalyzes the stereospecific synthesis of sn-1,2-diacylglycerol from sn-2-monoacylglycerol and long-chain fatty acyl-CoA. In order to study the regulation of MGAT, we developed a rapid assay that can be performed directly on permeabilized HA rat hepatocyte/hepatoma hybrid cells, a line that expresses levels of hepatic MGAT activity and a lipogenic program characteristic of fetal hepatocytes. In permeabilized HA cells, MGAT activity was proportional to the time of incubation and was highly dependent on added sn-2 monoacylglycerol and palmitoyl-CoA. The apparent Km values were 16.6 and 12.7 microM for palmitoyl-CoA and 2-monooleoylglycerol, respectively. Activity was low with the 1(3)- and sn-2-ether analogs of monooleoylglycerol, supporting the conclusion that the cells express the hepatic isoenzyme of MGAT. MGAT activity increased directly with cell density and was unrelated to the number of days in culture. Long-term incubation (2-4 days) of HA cells with various hormones (including triiodothyronine, human placental lactogen, epidermal growth factor, glucagon and growth hormone) showed that only a combination of dexamethasome and insulin resulted in significantly decreased MGAT activity. None of these hormones affected MGAT activity in short-term (0.5-4 h) incubations. These studies suggest that the developmental decline in rat hepatic MGAT activity may be regulated by glucocorticoids and insulin, hormones that increase during and after the second postnatal week. PMID- 8418889 TI - Characterization of the neutral pH-optimum sphingomyelinase from rat brain: inhibition by copper II and ganglioside GM3. AB - A neutral pH-optimum sphingomyelinase (N-SMase), solubilized from rat brain membranes, was characterized with respect to metal and membrane lipid effects. Chromatofocusing chromatography, which separates proteins according to pI, showed two N-SMase activities. One eluted at pH 4.7 and the other required 0.4 M NaCl before elution. Kinetically, the two preparations appeared similar. The N-SMase eluting at pH 4.7 was most extensively studied here. Of the phospholipids studied, only phosphatidylserine showed any influence on N-SMase and that was to increase its activity by as much as 50%. Neither serine nor phosphatidic acid had any effect. Of the cations tested, none was able to replace Mg2+ as a required activator. However, it was found that several metals were inhibitory, with Cu2+ being most effective (IC50 = 5 microM). Gangliosides, particularly the monosialoganglioside, GM3 (IC50 approximately 50 microM), inhibited N-SMase. Other glycolipids showed little effect on activity, even the immediate precursor to GM3 - lactosylceramide. The ganglioside sugar, N-acetylneuraminic acid, also had no effect on N-SMase activity. None of these inhibitors affected the acidic pH-optimum sphingomyelinase. Other sphingolipid compounds such as ceramide - the enzymatic product - and sphingosylphosphorylcholine (lysosphingomyelin) showed no capacity to inhibit N-Smase, implying that the enzyme may have a selective substrate-binding site. PMID- 8418890 TI - Reversible and irreversible inactivation of preformed pulmonary surfactant surface films by changes in subphase constituents. AB - Several mechanisms for surfactant inactivation have been reported. In this study, we have measured the reversibility of surfactant inactivation caused by various lipid or protein constituents of plasma or by pH changes. A surfactant of bovine origin was studied in a pulsating surfactometer either when surfactants were premixed with different serum constituents (inactivators) or when inactivators were introduced into subphase fluid surrounding surfactant films formed at an air liquid interface. Subphase exchanges with sodium bicarbonate or sodium borate raised pH and raised minimal surface tensions either when premixed with surfactant or when introduced with saline subphase beneath a preformed surfactant surface film. The pH effects on surfactant film function were reversible for sodium bicarbonate but not for sodium borate when the subphase with bicarbonate or borate was replaced with saline. Lipids (platelet-activating factor or lysophosphatidylcholine) had non-reversible effects on preformed films. Proteins (fibrinogen or C reactive protein) had reversible effects at low concentrations, but reversibility was less evident at high concentrations. Effects with whole serum were non-reversible at low protein concentrations (0.5 mg/ml). These results add evidence that surfactant inactivation can be caused by multiple mechanisms, both reversible and irreversible. PMID- 8418891 TI - Effects of high-density lipoprotein particles containing apo A-I, with or without apo A-II, on intracellular cholesterol efflux. AB - Previous reports have shown a differential effect of high-density lipoprotein (HDL) particles which contain apolipoprotein (apo) A-I without apo A-II (Lp A-I) and particles containing both apo A-I and apo A-II (Lp A-I/A-II) on cholesterol efflux from the mouse adipocyte cell line Ob1771, with Lp A-I and Lp A-I/A-II being active and inactive cholesterol efflux promotors, respectively. The present study was conducted to examine the roles of these two populations of apo-specific HDL particles on reverse cholesterol transport from cholesterol-loaded human skin fibroblasts and bovine aortic endothelial cells. The ability of HDL particles to remove intracellular cholesterol was tested by measuring depletion of the substrate pool for acylCoA:cholesterol acyltransferase (ACAT) and efflux of newly synthesized cholesterol, while removal of plasma membrane cholesterol was assessed by measuring efflux of [3H]cholesterol from prelabeled cells. Lp A-I and Lp A-I/A-II isolated from HDL2, HDL3 or plasma by immunoaffinity techniques each decreased esterification of cholesterol by both fibroblasts and endothelial cells. A mixture of Lp A-I and Lp A-I/A-II isolated from HDL3 decreased cholesterol esterification by fibroblasts in an additive manner, thus demonstrating that Lp A-I/A-II did not inhibit Lp A-I-mediated cholesterol efflux. Both Lp A-I and Lp A-I/A-II promoted efflux of sterol newly synthesized by fibroblasts, and no significant differences were observed between the apo specific particles. Apo-specific particles were also similarly effective at preventing the accumulation of LDL-derived cholesterol in cholesterol-depleted fibroblasts. Efflux of [3H]cholesterol from plasma membranes was stimulated to similar extents by Lp A-I and Lp A-I/A-II isolated from either HDL2, HDL3 or plasma. Thus, the apo-specific HDL particles Lp A-I and Lp A-I/A-II are both effective promoters of cholesterol efflux from fibroblasts and aortic endothelial cells. PMID- 8418892 TI - Relationship between cellular ATP, potassium, sodium and magnesium concentrations in mammalian and avian erythrocytes. AB - Intracellular K+/Na+ ratios of erythrocytes of various mammalian species are known to differ markedly. We have measured ATP, K+, Na+, Mg2+, H2O contents of erythrocytes of twelve mammalian and three avian species. Our results indicate that the intracellular ATP concentration in erythrocytes of different species is in close positive correlation with the K+/Na+ ratios (linear correlation coefficient, r = 0.852). Furthermore, ATP levels in erythrocytes of individual sheep with different potassium concentrations correspond with their K+/Na+ ratios (r = 0.747). Intracellular magnesium concentrations also correlate with ATP concentrations in erythrocytes of different species (r = 0.629) and in different sheep (r = 0.549). PMID- 8418893 TI - Identification of heterotrimeric and low molecular weight GTP-binding proteins in rabbit skeletal muscle longitudinal sarcoplasmic reticulum. AB - Direct photoaffinity labeling of proteins of longitudinal sarcoplasmic reticulum (LSR) of rabbit skeletal muscle with [32P]GTP revealed GTP-binding proteins of about 52, 45 and 30 kDa. ADP-ribosylation with [32P]NAD in the presence of cholera toxin (CTX) or pertussis toxin (PTX) indicates the existence of a CTX substrate (about 45 kDa); no PTX substrates were observed. Western blots of LSR probed with RM/1, an antiserum against a decapeptide from the C-terminus of Gs alpha, showed an immunoreactive band at about 45 kDa. [32P]GTP overlays of Western blots of LSR showed a heavily-labeled protein of about 29 kDa and one or more additional slightly smaller proteins that were more weakly labeled. When LSR was subjected to mild trypsin hydrolysis, the Western blot overlay revealed three bands at about 23, 25 and 29 kDa. Western blots of LSR proteins showed no significant immunoreactivity with the anti-(pan)-ras monoclonal antibodies 142 24E05 and Ras 11. ADP-ribosylation of LSR with [32P]NAD in the presence of C3 exoenzyme of Clostridium botulinum yielded a labeled band at about 23 kDa. Our results indicate the presence in rabbit LSR of a Gs alpha, the absence of Gi and G(o), and the presence of several low molecular weight GTP-binding proteins, distinct from p21 ras, one of which belongs to the rho or rac family. PMID- 8418894 TI - Strontium and barium induce exocytosis in electropermeabilized neutrophils. AB - Calcium, strontium and barium induced an exocytotic response in electropermeabilized rabbit neutrophils while magnesium was without any effect. The extent of enzyme release was found to depend upon the concentration of these cations. For all cations, an optimum concentration was found with the same maximum enzyme release. At concentrations higher than optimum a decrease in lysozyme release was observed. Efficiency to induce enzyme release was in the order: Ca2+ > Sr2+ > Ba2+. Enzyme release was significantly enhanced by guanosine 5'-[gamma-thio]triphosphate (GTP gamma S) resulting in a shift to the left of the dose/response curve. The enhancement by GTP gamma S was strongest with Ca2+, was less with Sr2+, and was very little with Ba2+. The time course of lysozyme release was the same for Ca2+, Sr2+, and Ba2+ in the presence and absence of GTP gamma S when suboptimal cation concentrations were used. A decrease in responsiveness to the effectors after electropermeabilization was observed with Ca2+, Sr2+ and Ba2+ in the presence and absence of GTP gamma S. The lysozyme release induced by the different cations was not inhibited by the protein kinase C inhibitor staurosporine and was slightly affected by pertussis toxin. Ca2+ and Sr2+, but not Ba2+, potentiated formyl-methionyl-leucyl-phenylalanine (fMet-Leu Phe) induced enzyme release in intact neutrophils. The divalent cation ionophore A23187 induced enzyme release in the presence of Ca2+ and Sr2+ but not in the presence of Ba2+. The results obtained with electropermeabilized neutrophils indicate that Sr2+ and Ba2+ can act as substitutes for Ca2+ in activating exocytosis, and that permeabilized neutrophils provide the best tool to investigate the effects of alkaline earth ions in exocytosis. PMID- 8418895 TI - A simple model for alanine metabolism in isolated rat hepatocytes. AB - A simple model describing reactions of alanine metabolism in isolated hepatocytes from fasted rats is proposed and applied to radioactive data obtained in experiments in which L-[1-14C]-, L-[2-14C]-, L-[3-14C]-, and L-[U-14C]alanine as well as L-alanine plus NaH14CO3 were used as substrates in parallel. Measurements of the rates of incorporation of the label into glucose and CO2 and of accumulation of [1-14C]pyruvate, [1-14C]lactate, [1-14C]alanine and [1 14C]glutamate plus [1-14C]glutamine from the different substrates used allows to calculate flux of alanine carbon through the various metabolic steps taken into account in the model. The validity of this model is indicated by the agreement found between calculations and measurement of the 14CO2 released from [1 14C]alanine as well as between the values of flux through pyruvate carboxylase calculated in two different ways. It is shown that the oxaloacetate synthesized by pyruvate carboxylase enters into the Krebs cycle and into the pathway of phosphoenolpyruvate synthesis in about equal proportions and that about 40% of the oxaloacetate synthesized as a result of alanine metabolism is derived from the Krebs cycle operation. These results, together with the conclusion that flux of alanine carbon through pyruvate dehydrogenase is negligible, are in agreement with known characteristics of hepatic alanine metabolism in the fasted state and, therefore, provide further evidence for the validity of the model proposed in the present study. PMID- 8418896 TI - Stimulatory effect of phospholipase A2 treatment on glucose utilization in human erythrocytes. AB - We examined whether modification of membrane phospholipids of human erythrocytes by hydrolysis with phospholipase A2 (PLA2 from bee venom) would affect glucose utilization, chosen as a typical model of intracellular metabolism, and, if so, intended to clarify the mechanism of the alteration of glycolysis. Treatment of erythrocytes with PLA2 induced a marked shape change (i.e., crenation) and significantly increased the rate of lactate production from glucose. Available evidence indicated that there is no relevance of this cell-shape change to the alteration of glycolysis. The lack of a detectable effect of papain treatment on glycolysis in PLA2-treated cells suggested that the increase in glycolysis by PLA2 treatment might not be caused by the conformational change of band-3 protein through modulation of membrane phospholipids. The result of the measurement of lactate production in the presence and absence of ouabain did not support the idea that hydrolysis of phospholipids by PLA2 treatment makes plasma membranes leaky to Na+ and consequently enhances glycolysis through activation of Na+/K(+) ATPase. The action of PLA2 on glycolysis was abolished by extraction of free fatty acids in the cell membrane with bovine serum albumin. Loading erythrocytes with free fatty acid (oleic acid, linoleic acid, or arachidonic acid) caused a significant increase in glycolysis. Analysis of glycolytic intermediates suggested that the enhancement of glycolysis was induced by activation of 6 phosphofructokinase. The data, thus, indicate that treatment of human erythrocytes with PLA2 significantly accelerates glucose utilization and suggest that the stimulation of glycolysis is caused by activation of 6 phosphofructokinase through liberation of free fatty acids of membrane phospholipids by PLA2. PMID- 8418897 TI - Characterization of phosphatidic acid phosphohydrolase in neutrophil subcellular fractions. AB - This investigation was designed to confirm the presence of PA phosphohydrolase in human neutrophils and to determine the distribution and characteristics of the enzyme in soluble and particulate subcellular fractions of disrupted neutrophils. Enzyme activity was detected in unseparated extracts of sonicated neutrophils. The majority of the recovered activity was recovered in a particulate fraction rich in neutrophil plasma-membrane markers; moderate levels (20%) of the total activity were recovered in the cytosol. While Mg2+ markedly potentiated the cytosolic but not the particulate activity, Ca2+ moderately inhibited both the cytosolic and particulate enzymes. The plasma-membrane-associated activity was absolutely dependent on detergent (0.5% Triton X-100) and displayed an apparent Km of 62 microM for phosphatidic acid. Enzyme activity was markedly inhibited by NaF, not influenced by excess glycerophosphate and slightly attenuated by propranolol, an inhibitor of PA phosphohydrolase in other systems. Preincubation of plasma membranes with N-ethylmaleimide at concentrations up to 25 mM had little effect on enzyme activity. However, activity in cytosolic and microsomal fractions of neutrophils were completely abolished by preincubation with N ethylmaleimide at concentrations of less than 5 mM. We conclude that neutrophils possess a potent PA phosphohydrolase localized in their plasma membranes. Metabolism of cellular second-messengers by this enzyme may exert a profound effect on the functions of stimulated neutrophils. PMID- 8418898 TI - Localization of 3 beta-hydroxysteroid dehydrogenase in the chicken ovarian follicle shifts from the theca layer to granulosa layer with follicular maturation. AB - 3 beta-Hydroxysteroid dehydrogenase (3 beta-HSD) catalyzes the conversion of pregnenolone to progesterone in the delta 4-3-ketosteroid metabolic pathway and dehydroepiandrosterone to androstenedione in the delta 5-3 beta-hydroxysteroid pathway. It has been suggested that small follicles of the chicken ovary that have not entered the follicular hierarchy metabolize steroids via the delta 5-3 beta-hydroxysteroid pathway, whereas preovulatory follicles that have entered the hierarchy metabolize steroids via the delta 4-3-ketosteroid pathway. Our objective was to localize 3 beta-HSD in follicles of the chicken ovary by immunocytochemistry using an anti-human placental 3 beta-HSD polyclonal antiserum to identify steroidogenic cells that convert pregnenolone to progesterone and/or dehydroepiandrosterone to androstenedione. Three groups of follicles of different maturities were examined: small follicles (1-10 mm in diameter and that have not entered the hierarchy), preovulatory follicles (10-35 mm in diameter and that have entered the hierarchy), and the most recent postovulatory follicle. Chicken ovaries were obtained 2 h after oviposition and fixed with Bouin's solution. Tissues were dehydrated with a series of ethanol, embedded in Paraplast (Brunswick Company, St. Louis, MO), and sectioned. Sections (4 microns) were immunostained for 3 beta-HSD with a Rabbit ExtrAvidin Staining Kit (Sigma Chemical Co., St. Louis, MO). 3 beta-HSD was localized in the single theca layer of cortical follicles (approximately 1 mm in diameter), which are still embedded in the stromal tissue, and in the theca interna and externa of other small follicles (< 10 mm in diameter). No immunoreactivity was observed in the granulosa layer of the majority of small follicles.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418899 TI - Effects of acute, intracerebroventricular administration of insulin on serum concentrations of luteinizing hormone, insulin, and glucose in ovariectomized lambs during restricted and ad libitum feed intake. AB - To test the hypothesis that the increase in pulsatile LH secretion associated with increased feed intake in growth-restricted lambs is due to elevated insulin concentrations, we evaluated the effects of intracerebroventricular (ICV) administration of insulin on patterns of LH in ovariectomized ewe lambs. After weaning (10 wk of age), 12 lambs were fed to maintain a mean body weight of 18.3 kg. At approximately 32 wk of age a permanent cannula was inserted into the lateral ventricle of each lamb. For the first experiment, animals received 3 ICV injections of either 500 ng (n = 6) or 500 micrograms (n = 6) insulin. Blood samples were collected every 10 min for 8 h, with animals receiving injections at 2, 4, and 6 h. Patterns of LH during the 2-h preinjection period were compared to those in the subsequent 2-h periods following each insulin injection. Insulin did not affect mean LH, LH pulse frequency, or LH pulse amplitude. Only the 500 micrograms injections increased (p < 0.0001) peripheral insulin and decreased (p < 0.001) peripheral glucose. The experimental protocol was repeated during a second experiment conducted after 2 wk of ad libitum feeding, when animals weighed an average of 21.4 kg. Increased feed intake was associated with increases in mean LH, LH pulse frequency, and insulin (p < 0.05). Both doses of insulin decreased (p < 0.01) mean LH and LH pulse frequency. The 500-micrograms injections increased (p < 0.0001) peripheral insulin and decreased (p < 0.001) glucose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418900 TI - Identification of a 30,000 M(r) polypeptide secreted by cultured ovine granulosa cells and luteal tissue as a tissue inhibitor of metalloproteinases. AB - Previous studies have revealed that the gonadotropin surge initiates, via transcriptional mechanisms, synthesis and secretion of a 30,000 M(r) polypeptide by the granulosa cells of ovine preovulatory follicles. This polypeptide also appears to be secreted by luteal tissue and has been tentatively identified as a tissue inhibitor of metalloproteinases (TIMP; 68% NH2-terminal amino acid sequence identity to a human TIMP). Therefore, the objective of the present study was to confirm that TIMP is produced by granulosa cells and luteal tissue in the ewe. A series of experiments was conducted to determine whether the 30,000 M(r) polypeptide secreted by granulosa cells and luteal cells is similar to TIMP in biochemical properties (degree of N-linked glycosylation), in biological activity (as ascertained by gelatin zymographic analysis), and in immunoreactivity (as ascertained by Western blot analysis). Incubation of granulosa cells or luteal cells with tunicamycin revealed that the 30,000 M(r) polypeptide is glycosylated. The form lacking N-linked chains had an M(r) (approximately 20,000) similar to that of the unglycosylated form of TIMP in other species. Gelatin zymographic analysis detected significant metalloproteinase inhibitor activity associated with polypeptides of M(r) approximately 21,000 and 30,000 secreted by granulosa cells and luteal cells. Northern hybridization of granulosa cell RNA and Day 10 luteal RNA with an anti-sense TIMP oligonucleotide probe detected an approximately 900-base transcript, which is similar in size to that reported for TIMP mRNA in other species. Finally, Western blot analysis with a rabbit anti human TIMP antiserum detected immunoreactive polypeptides of M(r) 30,000 secreted by granulosa cells and luteal cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418901 TI - How does daily treatment with human chorionic gonadotropin induce superovulation in the cyclic hamster? AB - Daily s.c. injection of 2.0 IU hCG per day, begun on Day 1 of the cycle (estrus), results in hamsters ovulating 20.7 +/- 0.7 eggs instead of the normal number of 13.3 +/- 0.5 (SEM). This is associated with a reduced rate of follicular atresia so that more of the 10 developing follicles per ovary (large preantral stages) normally recruited on Day 1 of the cycle mature and go on to ovulate. The hCG treated follicles were larger than control follicles, but contained similar amounts of DNA/follicle; increased size of the antral cavity accounted for their greater size. Moreover, DNA synthesis was significantly reduced in the hCG follicles on Days 2 and 4. Thecal vascularity as judged by the number of red blood cells retained in the theca or microsphere uptake by follicles indicates that on Day 2, thecal blood flow was significantly lower in the hCG-treated animals than in controls. On the other hand, after hCG treatment begun on Day 1, serum levels and in vitro incubation of individual follicles revealed that on Day 2 and beyond, androstenedione (A) and estradiol (E2) levels were elevated. After hCG treatment, the elevated serum E2 correlated with reduced serum LH on Days 3 and 4 whereas FSH was unaffected. To study in vitro steroid accumulation, the 10 largest follicles (the developing follicles) were dissected from alternate left and right ovaries from control and hCG-treated animals and incubated individually, and their histology was then compared with the steroid profiles. Accumulation of A and E2 was significantly greater in the hCG-treated follicles than in controls in a 1-h basal incubation and after the addition of 50 ng LH. Progesterone accumulation usually did not differ between the control and hCG treated follicles. Early stage 1 atretic follicles (judged by histology) were still capable of producing A and E2 in vitro, comparable to control follicles; but, as atresia progressed, the follicles synthesized only progesterone. This is consistent with the temporal pattern previously observed in a model of induced follicular atresia in the hamster [Greenwald, Biol Reprod 1989; 40:175-181]. It is concluded that superovulation resulting from hCG injections is due to thecal production of androgens from follicles normally destined for atresia. For the untreated cyclic hamster, the critical time for thecal androgen production is the first 2 days of the cycle. The aromatizable androgens are then converted into estrogens, which in turn may maintain the microenvironment of the antral cavity, which is essential for viability of the granulosa cells.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418902 TI - Colchicine disrupts the cytoskeleton of rat testis seminiferous epithelium in a stage-dependent manner. AB - Sertoli cell microtubules play an important role in the process of spermatogenesis. We investigated the effects of colchicine, a microtubule disrupting agent, on the seminiferous epithelium. Rats were injected intratesticularly with 0.004-40 micrograms colchicine/testis. Colchicine had a dose-related effect on seminiferous tubule fluid secretion and completely blocked secretion at a dose of 40 micrograms colchicine/testis. Colchicine also resulted in a dose-related decrease in testes weight 2 and 8 wk after injection. When 40 micrograms colchicine/testis was used, testis morphology showed a time-dependent increase in the incidence of sloughing over a time course of 1, 3, 6, and 16 h. Quantitative analysis demonstrated that stage IX-XIV seminiferous tubules were most sensitive to sloughing. Changes in the distribution of tubulin immunostaining within Sertoli cells occurred preferentially in stage VII-VIII seminiferous tubules, which were most resistant to sloughing. In addition, colchicine resulted in disruption of vimentin filaments in stage IX-XIV seminiferous tubules, which correlated with the stage-dependent sensitivity of sloughing. We propose that the stage dependence of colchicine-induced effects reflects the dynamic and stage-dependent role of microtubules in spermatogenesis. Furthermore, cellular structures other than microtubules, such as vimentin filaments, may be important for maintaining the structural integrity of the seminiferous epithelium. PMID- 8418903 TI - An update on the roles of inhibin, activin, and follistatin as local regulators of folliculogenesis. AB - The local actions of the inhibin-related peptides inhibin, activin, and follistatin in folliculogenesis, luteinization and atresia are reviewed. On the basis of in vitro data, it is concluded that (a) inhibin has a paracrine action positively regulating LH-induced androgen production by theca cells in addition to a peripheral role regulating FSH secretion; (b) activin has a negative paracrine action on LH-induced androgen production by theca cells; (c) activin, either alone or with FSH, has an autocrine action on granulosa cells, promoting differentiation during the preantral and early antral stages of folliculogenesis and preventing premature luteinization in the later stages of antral follicle development, leading overall to promotion and maintenance of the folliculogenic state of the follicle; and (d) follistatin modulates granulosa cell function in favor of luteinization or atresia, and its mechanism of action involves neutralizing the actions of activin by its binding properties and by a direct action of follistatin on progesterone metabolism by granulosa cells. A hypothesis for a role of activin in the acquisition of responsiveness of granulosa cells of preantral follicles to FSH is proposed. PMID- 8418904 TI - Evidence that 68-kilodalton and 54-51-kilodalton polypeptides are components of the human sperm fibrous sheath. AB - This study characterizes a common antigen recognized by two monoclonal antibodies (mAbs) that immunoreact with the principal piece of the human sperm flagellum. By means of immunofluorescence microscopy, mAbs S69 (IgM) and S70 (IgG1) (WHO Task force nomenclature) were observed to bind to the principal piece of methanol- or detergent-permeabilized human spermatozoa, but did not react with live swimming spermatozoa as assessed by immunofluorescence microscopy. Faint immunofluorescence was also seen on the connecting piece in approximately 40% of the spermatozoa. Immunoreactivity in both regions was resistant to sequential extraction with Triton X-100, sodium thiocyanate, and urea. Pre-embedding electron microscopic immunogold labeling of ejaculated spermatozoa with mAb S69 showed gold particles located on the fibrous sheath. Immunoreactive peptides of 68, 53, and 45 kDa were recognized by both S69 and S70 mAbs on immunoblots of nonreduced human sperm extracts, while a 68-kDa band and a strongly immunoreactive triplet from 54 to 51 kDa were recognized in reduced sperm extracts. Human fibrous sheaths were isolated by differential solubilization and centrifugation and characterized by transmission electron microscopy. The 68-kDa and 54-51-kDa bands were enriched and found to be major polypeptides in the isolated fibrous sheath fraction. These results suggest that the S69/S70 antigen, which we term SP (sperm protein) (68 kDa, 54-51 kDa), is a component of the human fibrous sheath. PMID- 8418905 TI - Effects of cytokines on prostaglandin production and steroidogenesis of incubated preovulatory follicles of the rat. AB - Cytokines, leukocyte-derived peptide regulators of inflammation and other lympho hematopoietic processes, have been implicated in ovarian physiology on the basis of findings of cytokines in follicular fluids and the presence of leukocytes in ovarian tissue. During the dramatic tissue remodelling at ovulation, several inflammatory mediators play pivotal roles in the occurrence of follicular rupture, but no data exist regarding the involvement of cytokines in this process. In the present study we have examined the effects of three different lymphocyte- and macrophage-derived cytokines--tumor necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1 beta), and interleukin-2 (IL-2)--on the production of ovulatory mediators such as prostaglandins and progesterone in the preovulatory follicle of the eCG/hCG-primed immature rat. Preovulatory follicles were incubated in the presence of human recombinant cytokines for up to 24 h, and the concentrations of prostaglandins and steroids in the incubation medium were measured by RIAs. TNF alpha and IL-1 beta stimulated production of prostaglandin E and prostaglandin F2 alpha in a dose-dependent manner during a 24-h incubation. Synthesis of prostacyclin was also stimulated by TNF alpha and IL-1 beta, as indicated by high levels of its stable metabolite 6-keto-prostaglandin F1 alpha. Time-course studies showed that a major portion of the prostaglandins were produced between 12 and 24 h. IL-2 was without effect on these parameters. TNF alpha and IL-1 beta also dose-dependently increased the production of progesterone and this was not inhibited by indomethacin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418906 TI - Comparison of follicular estrogen receptors in rat, hamster, and pig. AB - The hypophysectomized or intact hamster--unlike the rat--does not respond to estrogen by increasing the number of large antral follicles. In the pig, estrogen inhibits progesterone synthesis by granulosa cells from large follicles for 36 h before progesterone accumulation is enhanced. The objective of this study was to determine whether these divergent responses of the three species to estrogen are related to differences in follicular estrogen receptors. For the rat antral follicle, the equilibrium dissociation constants (Kds), expressed as nmol/L, were 0.55 +/- 0.07 in the cytosolic fraction and 0.46 +/- 0.11 in the nuclear fraction; for the hamster and pig follicles, the respective Kds were 1.08 +/- 0.04 and 0.94 +/- 0.11, and 1.16 +/- 0.23 and 1.09 +/- 0.17, respectively. The cytosolic and nuclear Kds for the rat follicle were statistically different from those of the other two species, but this is most likely not of biological consequence. Comparison of the maximal numbers of binding sites (NBSmax) in the cytosolic fractions of the antral follicles, expressed as fmol/mg protein, indicated a relationship of hamster > rat = pig; whereas in the nuclear fraction, with the NBSmax expressed as fmol/mg DNA, the relationship was hamster > pig > rat. The total number of estrogen receptors (sum of the NBSmax of cytosolic and nuclear fractions) was approximately tenfold higher in the hamster than in the rat. Thus, the failure of hamster large follicles to respond in vivo or in vitro to estrogen cannot be explained on the basis of lack of estrogen receptors.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418907 TI - Developmental changes in mouse placental cells from several stages of pregnancy in vivo and in vitro. AB - Developmental changes in mouse placentae from the 6th to the 18th day of pregnancy were studied in vivo and in vitro. Placental volume increased from the 6th to the 18th day of pregnancy; however, the total number of cells per placenta reached a plateau on the 14th day. Decidual cells were predominant in the placenta on the 6th day. Placentae obtained from the 10th to the 18th day contained decidual cells, trophoblastic (labyrinth and spongiotrophoblast) cells, and trophoblast giant cells. Decidual cells increased in number from the 6th to the 10th but decreased on the 14th day, whereas trophoblastic cells increased linearly until the 14th day. Two types of placental cells were distinguished in vitro: small fibroblast-like cells and large flattened cells containing 2-3 nuclei. The large cells reacted to anti-desmin antibody, indicating their decidual character. The small cells reacting to anti-keratin antibody appeared to be trophoblastic cells. Decidual cells from all days of gestation were nonproliferative, regressing with time in culture. 17 beta-Estradiol (E, 10(-9) and 10(-8) M), progesterone (P, 10(-10), 10(-9), and 10(-8) M), and a combination of E and P (10(-9) M each) stimulated proliferation of the trophoblastic cells only from the 6th and the 10th days. Keoxifene (2 x 10(-7) M), but not tamoxifen, significantly inhibited the E-induced proliferation of the trophoblastic cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418909 TI - Immunocytochemical localization of progesterone receptor in the reproductive tract of adult female rats. AB - The distribution of the progesterone receptor (PR) was investigated immunocytochemically in female reproductive tracts of rats during the estrous cycle and early pregnancy through use of an anti-PR monoclonal antibody. PR was localized predominantly in the nuclei of epithelial, stromal, and muscle cells in the uterus and vagina during the estrous cycle. In the uterus, the nuclei of epithelial cells were stained intensively at diestrus, while the PR staining of the stromal cells was more intense at proestrus than at any other stage of the cycle. PR expression during the cycle in muscle cells of the myometrium was similar to that in the endometrial stromal cells. In the vagina, however, PR expression during the cycle was approximately the same among epithelial, stromal, and muscle cells, the nuclei of which were stained deeply at proestrus. Ovariectomy at various stages of the cycle altered the PR expression appearing in the uterus and vagina during the cycle. In ovariectomized rats, estrogen increased the PR immunoreaction of various types of cells examined in the uterus and vagina except for the uterine epithelial cells. The reaction of these uterine epithelial cells was decreased by estrogen but was increased by progesterone given after estrogen; however, progesterone given alone reduced the reaction. In the epithelial and stromal cells of the uterus, intensity of the staining was increased after mating, reaching maximum on Day 3 of pregnancy, and then decreased on Day 4 (day of implantation), while in epithelial and stromal cells of the vagina the staining remained weak during early pregnancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418908 TI - Seasonal changes in the relationships between secretion of gonadotropin-releasing hormone, luteinizing hormone, and testosterone in the ram. AB - In this study we examined the relationship between GnRH, LH, and testosterone (T) in intact rams during the nonbreeding season, the breeding season, and the transition between breeding and nonbreeding season. Blood was collected from the hypophyseal portal and jugular veins at 10-min intervals for 12 h from 3 groups of rams in May, September, and March. Individual LH pulses in May and September occurred at relatively regular intervals whereas in March, groups of pulses were separated by relatively long intervals without pulses. No significant effect of season on GnRH pulse parameters was found, but there was a significant effect of season on LH (p < 0.04) and T (p < 0.001) pulse amplitude and on mean T concentrations (p < 0.001). LH pulse amplitude was highest in March, and T pulse amplitude was highest in September. Simple indices of pituitary and testicular responsiveness were obtained by calculating the ratios of LH to GnRH and T to LH. The ratio of LH to GnRH pulse amplitude was significantly higher in March than in September (p < 0.01) or May (p < 0.05). The ratio of T to LH pulse amplitude was highest (p < 0.01) in September. Release of LH in response to exogenous GnRH also varied significantly (p < 0.01) with month, being higher in March than in September (p < 0.01) or May (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418910 TI - Carl G. Hartman Award. Neena B. Schwartz. PMID- 8418911 TI - SSR Research Award. Jerome F. Strauss III. PMID- 8418912 TI - SSR Distinguished Service Award. Luigi Mastroianni, Jr. PMID- 8418913 TI - New Investigator Awards. PMID- 8418914 TI - The Society for the Study of Reproduction: 25 years in retrospect. PMID- 8418916 TI - Localization of transforming growth factor beta 1 and beta 2 during testicular development in the rat. AB - The transforming growth factor beta s (TGF beta s) affect the metabolic activities of the somatic cells of the testis. Sertoli cells, peritubular/myoid cells, and germ cells contain mRNA for TGF beta 1 and/or TGF beta 2. We have used immunohistochemical techniques to determine, in vivo, when TGF beta 1 and TGF beta 2 are present in the rat testis during development and have identified the precise localization of these growth factors. The most pronounced changes in TGF beta immunoreactivity occurred during spermatogenesis. TGF beta 1 predominated in spermatocytes and early round spermatids, but as the spermatids elongated around stages VIII-IX of the cycle, the TGF beta 1 levels declined. TGF beta 2 was undetectable in spermatocytes and early round spermatids, but as spermiogenesis progressed, around stages V-VI, the spermatids rapidly acquired TGF beta 2. The intense staining for TGF beta 2 was maintained as the spermatids elongated. TGF beta 1 immunoreactivity was detected in Sertoli cells throughout testicular development. TGF beta 2 was found in fetal Sertoli cells, but became undetectable rapidly after birth. In fetal animals the Leydig cells contained TGF beta 1 and TGF beta 2; after birth TGF beta 1 persisted whereas TGF beta 2 became undetectable in the Leydig cells. Prior to puberty, TGF beta 1 and TGF beta 2 were absent in a portion of the Leydig cells; when the adult stage was reached, TGF beta 1 was no longer detectable and TGF beta 2 staining was faint to absent. In conclusion, our novel findings show that TGF beta 1 and TGF beta 2 are present in vivo in testicular cells at clearly defined stages of their differentiation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418915 TI - A murine model of preterm labor: inflammatory mediators regulate the production of prostaglandin E2 and interleukin-6 by murine decidua. AB - Inflammatory processes may be important in the initiation and propagation of uterine contractions and preterm labor in human pregnancies. Recently, a murine model of preterm labor has been described. The purpose of our study was to determine whether murine decidua responds to inflammatory mediators, such as lipopolysaccharide (LPS) and the inflammatory cytokines interleukin (IL)-1 beta and tumor necrosis factor (TNF). Allogeneic pregnant mice (C3H/Hen females mated with C57/B6 males) were killed at 12-14 days of pregnancy, decidual tissue was isolated, and explants were placed on the polycarbonate membrane of Costar Transwell inserts. Initial validation studies of this explant system, including biochemical and histologic evaluations, indicated that the decidual tissue remained intact, viable, and responsive to IL-1 beta for at least 5 days in explant culture. Treatment of murine decidual explants with LPS, IL-1 beta, and TNF resulted in significant increases in the production of prostaglandin E2 (PGE2) and IL-6. Thus the regulation of PGE2 and IL-6 production from murine decidua by LPS and inflammatory cytokines is similar to findings previously reported for human decidua. Our findings are consistent with the view that the pathophysiology of infection-induced preterm labor in the mouse may be similar to that in human pregnancy and supports the continued development of murine models of preterm labor. PMID- 8418917 TI - Comparison of functional response of rat, macaque, and human ovarian cells in hormonally defined medium. AB - A serum-free medium has been developed which supports in vitro function by ovarian cells derived from rat, monkey, and human tissue. This granulosa cell medium (GCM) consists of Dulbecco's Modified Eagle's Medium: Ham's F-12 medium (1:1, v:v) supplemented with insulin, transferrin, aprotinin, selenium, fibronectin, penicillin, and streptomycin. Ovarian cells from three species were compared: rat, macaque, and human. Four types of ovarian cultures were examined: 1) purified granulosa cell cultures and 2) co-cultures containing granulosa-theca stroma cells, 3) luteal cells, and 4) granulosa-lutein (harvested from in vitro fertilization cultures) cells. Each cell type was characterized by its response to FSH or hCG when cultured in GCM. Morphologic responses to FSH were observed in GCM in rat granulosa and granulosa-theca-stroma cell cultures, macaque and human granulosa-lutein cells, and human granulosa-theca-stroma cell cultures. The FSH stimulated cells retracted and became rounded, leaving long intercellular connections. Luteal cells did not retract in response to FSH, and the cells remained firmly attached to the fibronectin matrix. Steroidogenic regulation of the GCM-cultured ovarian cells was monitored following stimulation of the cultures with FSH. The ability of the cells to aromatize testosterone was first examined. Rat granulosa cell cultures and granulosa-theca-stroma cell cultures, macaque granulosa-lutein cell cultures, and human granulosa-theca-stroma cell cultures all accumulated estradiol when given FSH and testosterone for 48 h. Moreover, these cell types as well as human luteal cells were able to metabolize 25-hydroxy [1,2-3H]cholesterol to various steroid metabolites. The data indicate that GCM supports normal granulosa cell morphologic response to FSH.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418918 TI - Immunolocalization of oviductin in endocytic compartments in the blastomeres of developing embryos in the golden hamster. AB - We have previously localized an antigen of oviductal origin in the zona pellucida of superovulated hamster ova. This antigen is a high-molecular-weight glycoprotein that is secreted by the nonciliated secretory cells of the oviduct and later is transferred to the zona pellucida of the oocyte during oviductal transit. This glycoprotein is rich in N-acetyl-D-galactosamine residues and has been designated Hamster Oviductin-1. In the present study, we have examined the intracellular localization of the oviductin in the blastomeres of developing embryos of the golden hamster. Seventeen cycling females were used for the localization and detection of the oviductin in oviductal oocytes and early embryos. Thin sections of hamster 1-cell oocytes, and 2-cell, and 8-cell embryos, embedded in Lowicryl and then incubated with the monoclonal antibody against the oviductin, revealed a homogenous distribution of antigenic sites in the matrix of the zona pellucida. While immunogold labeling was completely absent in the cytoplasm of 1-cell ova prior to fertilization, labeling was found associated with coated pits and coated vesicles and with flocculent material in the perivitelline space of fertilized eggs. In addition to these labeled endocytic structures, many endosomes, multivesicular bodies, and secondary lysosomes were also found to be labeled heavily in the cytoplasm of developing embryos. Our results indicate that following the transfer of Hamster Oviductin-1 to the zona pellucida of oocytes during transit in the oviduct and subsequent to fertilization, some of the oviductin associated with the zona pellucida appears to be internalized by blastomeres of the embryo and further processed through the endosomal/lysosomal pathway. PMID- 8418919 TI - Perturbations in the model of estrogen receptor regulation of gene expression. AB - Estrogen receptors are key elements in the mechanisms of action of estrogenic hormones. Models of how estrogens and their receptors interact and subsequently modify gene expression should be reevaluated to explain new data currently available. The following review discusses nuclear localization, DNA binding, and protein structural changes of the estrogen receptor induced by estrogen binding. We also discuss how these phenomena relate to the induction of changes in gene expression. PMID- 8418920 TI - Changes in rat uterine estrogen receptor messenger ribonucleic acid levels during estrogen- and progesterone-induced estrogen receptor depletion and subsequent replenishment. AB - The overall objectives of this study were to determine whether the rapid decrease in estrogen receptor (ER) binding in the rat uterus after an injection of estradiol and subsequent recovery of ER levels was accompanied by similar changes in ER mRNA levels. Furthermore, the effect of progesterone administered under conditions known to decrease ER binding, in the rat uterus, on ER mRNA levels was also investigated. Ovariectomy for 14 days brought about a 3-fold increase in rat uterine ER mRNA levels, and these elevated levels were decreased by a 3-day treatment with 2 micrograms of estradiol in ethanol/saline injected i.p. In the ovariectomized rat, 1 and 5 micrograms of estradiol brought about small but significant decreases in ER mRNA levels in 1 h, which did not parallel the rapid decrease in ER binding at that time reported earlier. The 10-micrograms dose of estradiol brought about a bigger decrease in ER mRNA levels. In the ovariectomized rat primed with estradiol (2 micrograms/day in ethanol/saline), the administration of 2 micrograms of estradiol brought about no change in uterine ER mRNA levels at 6 h as compared to the 1-h time point if the values were not corrected for beta-actin, which was significantly increased at 6 h. A dramatic increase in ER mRNA levels 12 h after the estradiol injection preceded the increase in ER binding observed at 18 h. Progesterone (0.8 mg/kg body weight [BW]) in the absence of estrogen priming brought about minimal but significant inhibition of ER mRNA levels 12 and 18 h after administration, with no effects at 1 and 6 h. In the presence of estrogen priming, the 0.8-mg/kg BW dose of progesterone did not cause any changes in ER mRNA levels beyond those brought about by estrogen alone after 1 h, even though it has been shown to significantly decrease ER binding. This was also true when a larger dose of progesterone (2.0 mg/kg BW) was used, a dose that decreases ER binding to a significantly greater extent than the 0.8-mg/kg BW dose. However, the 4.0-mg/kg BW dose of progesterone decreased ER mRNA levels. Thus a single injection of estradiol appears to cause a decrease in ER binding primarily by accelerated receptor processing and degradation with small changes in ER mRNA within the first hour. A significant increase in uterine ER mRNA at 12 h precedes increased ER binding at 18 h. This indicates new synthesis of ER during the recovery period.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418921 TI - Determination of water permeability coefficient for human spermatozoa and its activation energy. AB - Four experiments were conducted to determine the permeability coefficient of human sperm to water (Lp) and its activation energy (Ea). Critical tonicity (tonicity at which 50% of the cells swell and lyse) was determined by equilibrating sperm to 22 degrees C (experiments 1a and 1b), 30, 22, 8, or 0 degrees C (experiment 2a), and 0, -1, -3, -5, or -7 degrees C (experiment 2b) and then exposing them to various hypotonic media (215-3 mOsm). For Lp determination, sperm were equilibrated to 30, 22, 8, or 0 degrees C (experiment 3a), 8, 0, or -3 degrees C (experiment 3b), and -1, -3, -5, or -7 degrees C (experiment 3c), and then were exposed for increasing times to hypotonic (40 mOsm) media. Activation energies were calculated from the results of the latter experiments (experiment 4). Results indicate a temperature-dependent (p < 0.05) critical tonicity, with sperm exhibiting an increased membrane fragility at 8, 0, and -7 degrees C, relative to 30, 22, -1, -3, or -5 degrees C (67.5 +/- 2.4, [mean +/- SEM], 62.7 +/- 2.3, and 61.9 +/- 3.7 mOsm vs. 57.4 +/- 3.4, 57 +/- 1.2, 54.8 +/- 3.4, 60.1 +/- 5.3, and 59.8 +/- 5.2 mOsm, respectively). Human sperm have an Lp of 2.40 +/- 0.20 microns/min/atm at 22 degrees C and an Ea of 3.92 +/- 0.59 kcal/mol between 30 and -7 degrees C. The Ea for cells incubated at temperatures above 0 degrees C (3.92 kcal/mol) show an apparent discontinuity (p < 0.004) in water permeability in supercooled conditions (7.48 kcal/mol). These data suggest that 1) human sperm have a high Lp and low Ea, relative to other cell types, above 0 degrees C; and 2) this high Lp and its low Ea change significantly below 0 degrees C. PMID- 8418922 TI - Regional alterations in the levels of brain biogenic amines, glutamate, GABA, and GAD activity due to chronic consumption of inorganic arsenic in developing and adult rats. PMID- 8418923 TI - Toxicity studies of antiscalant agents using Arbacia punctulata gametes and embryos as test organisms. PMID- 8418924 TI - Evaluation of fenitrothion toxicity to Rana temporaria L. PMID- 8418925 TI - In vitro skin absorption of 14C-cyanuric acid in a simulated swimming pool. PMID- 8418926 TI - Estimation of hemosiderosis in seabirds and fish exposed to petroleum. PMID- 8418927 TI - Effect of cyclic aromatics on sodium active transport in frog skin. PMID- 8418928 TI - Lead, cadmium, nickel, and zinc contamination of ground water around Hussain Sagar Lake, Hyderabad, India. PMID- 8418929 TI - Toxicity of hexavalent chromium from aqueous and sediment sources to Pimephales promelas and Ictalurus punctatus. PMID- 8418930 TI - Effect of tributyltin on mortality and telson regeneration of grass shrimp, Palaemonetes pugio. PMID- 8418931 TI - Mercury concentrations in oysters from the coastline of Northern Territory, Australia. PMID- 8418932 TI - Acute toxicity of sulfide and lower pH in cultured rainbow trout, Atlantic salmon, and coho salmon. PMID- 8418933 TI - Analysis of selenium metabolites in human urine using ion exchange chromatography. PMID- 8418934 TI - Degradation of two acetanilide herbicides in a tropical soil. PMID- 8418935 TI - Effect of moisture regime, temperature, and organic matter on soil persistence of carbosulfan. PMID- 8418936 TI - Monitoring movement of fenamiphos through soil water in peach orchards using quantitation by gas chromatography/mass spectrometry. PMID- 8418937 TI - Effect of nematicides, Telone II and Vorlex, on microflora and nitrification in tobacco soil. PMID- 8418938 TI - Effects of carbofuran and the corn rhizosphere on growth of soil microorganisms. PMID- 8418939 TI - Residue levels of carbendazim in opium poppy (Papaver somniferum). PMID- 8418940 TI - Effects of omega-chloroacetophenone (CN) vapor inhalation on pulmonary immune system of mice. PMID- 8418941 TI - Influence of pre- and simultaneous perfusion of ascending concentrations of lead on the effect of elevated calcium on digoxin-induced cardiac arrest in isolated frog heart. PMID- 8418942 TI - Organochlorine pesticides and polychlorinated biphenyls (PCBs) in human serum collected from the general population from Zagreb (1985-1990). PMID- 8418943 TI - Effect of aluminum on lipid peroxidation of cerebral hemisphere of chick. PMID- 8418944 TI - Spectroscopic studies on the effects of aluminum ion on calf-thymus DNA. PMID- 8418945 TI - A chronicle of DNA methylation (1948-1975). PMID- 8418946 TI - Structure, function and regulation of mammalian DNA methyltransferase. PMID- 8418947 TI - Regulation of de novo methylation. PMID- 8418949 TI - CpG islands. PMID- 8418948 TI - Effect of DNA methylation on the binding of vertebrate and plant proteins to DNA. PMID- 8418950 TI - Control of DNA methylation in fungi. PMID- 8418951 TI - Patterns of de novo DNA methylation and promoter inhibition: studies on the adenovirus and the human genomes. PMID- 8418952 TI - Methylation of cytosine influences the DNA structure. PMID- 8418953 TI - Effect of DNA methylation on dynamic properties of the helix and nuclear protein binding in the H-ras promoter. PMID- 8418954 TI - DNA methylation and embryogenesis. PMID- 8418955 TI - Epigenetic inheritance based on DNA methylation. PMID- 8418956 TI - DNA methylation and genomic imprinting in mammals. PMID- 8418957 TI - DNA methylation and cancer. PMID- 8418958 TI - Gene methylation patterns and expression. PMID- 8418959 TI - What do I want? A model for self discovery. PMID- 8418961 TI - The professional staff organization. PMID- 8418960 TI - Are contract management services a threat to hospital-based clinical engineers? PMID- 8418962 TI - Real-time flexible preventive maintenance scheduling. AB - There are still obstacles to overcome as we enter the programming phase of this project. We envision an automated system, similar to an expert system, that performs the interval/history analysis and makes the changes. Initially a field will need to be added to the inventory to denote whether a device belongs to one of the previously described groups that are exempt from interval changes. An intermediate step will be the formatting of a periodic report showing equipment that meets the change criteria as described in the two rules. For now, the actual changes would be reviewed and made by our management and technical staff. This report would be retained as documentation of the basis for each change, for our own benefit and to meet JCAHO requirements. We are still discussing whether the repair count should include all repairs (user error, abuse, unpredictable failure, etc.) or just those that are "significant and preventable" and could have been averted by PM. This is perhaps a question whose answer might vary from hospital to hospital, depending upon size and patient mix. With more emphasis being placed on process outcomes, on quality of work, and on getting the most benefit for our efforts, we believe our flexible, real-time PM scheduling program is a major step in the right direction. It is outcome-driven and it focuses resources where they are needed the most. PMID- 8418963 TI - Biomedical equipment considerations for aeromedical transports. AB - Due to the eight stresses of flight and Federal Aeronautics Administration (FAA) requirements, biomedical equipment that is utilized in aeromedical transports presents certain challenges that the biomedical department should be aware of. U.S. Air Force military studies of a large number of specific models are available through the government. This author recommends prepurchase flight tests and input from flight crews to ensure safe operation of any new equipment. The equipment should also be designed for air transport. Permanent pacemakers should be programmed to a non-atrial sensing mode or an asynchronous mode before the patient is on board the aircraft. Temporary pacers and automatic defibrillators should also be set to a mode where the vibrations of flight will not trigger any errant behavior. With the proper precautions, aeromedical transports will continue to be a rapidly growing transport system for both trauma patients and intrahospital transfers. With a little research, the biomedical engineer can also be a valuable asset to the ground support crew. PMID- 8418964 TI - A clinical engineering information system incorporating ECRI-HECS, HECSLINK, and dBase. PMID- 8418965 TI - Biometry with a video-genlock interface and a computer-based image-analyzing system: use as a TV-videopupillometer. AB - TV-videobiometry is frequently used in biomedical research. Usually, the video images are digitized by computer systems. In contrast, the technique described uses a device known as a video-genlock interface (genlock). The genlock mixes a computer picture with the picture of a video device (synchronization). This technique and the custom-made software provide the following advantages: programmable user interface, low-budget system, user-friendly, manual-based software package, precision, and usefulness in different videobiometric setups such as infrared videopupillometry, which is described. In the demonstrated videopupillometry, the area resolution is 0.0009 mm2 per pixel and the length resolution 0.03 mm per pixel side. The relative coefficient of variation for 20 repeated measurements of pupillary area lies between 0.15 and 0.35%. The mean error of three measurements of a model pupil is less than 1%. PMID- 8418966 TI - Manual verification of computer analysis of 24-hour esophageal motility. AB - The application of solid-state technology to intraesophageal pressure monitoring over an entire circadian cycle has resulted in large amounts of data that require computer analysis. Recently available commercial software has yet to be validated. The aim of this study was to compare the analysis of ambulatory esophageal manometry by an automated computer program with manual analysis and make the software modifications necessary to validate the automated system for clinical use. Computer-aided analysis of a large number of esophageal contractions recorded during ambulatory esophageal manometry was compared with manual analysis by four experienced physicians. Good correlations were found between manual and computerized measurements of contraction amplitude and duration (r = 0.99 and r = 0.73, respectively). Software modifications resulted in correct identification of 94% of contractions and correct classification of 93.3% of these waves as peristaltic or simultaneous. These results demonstrate that the evaluated program for automated analysis of ambulatory esophageal manometry is accurate and reliable for research and clinical applications. PMID- 8418967 TI - Adaptive compression of the ambulatory electrocardiogram. AB - Previous use of the MIT/BIH arrhythmia database, on analog tape, to investigate compression of ambulatory ECG data by average beat subtraction, residual differencing, and Huffman coding of the residuals had shown that with a quantization level of 35 mu V and a sample rate of 100 samples per second, it was possible to store ECG data with average data rates of 174 bits per second (bps), but because of the variation in ECG signals, data rates for different records ranged from 144 bps to 230 bps. In a practical storage system, it is desirable to fix the maximum data rate and store data with a minimum of distortion. For this study the previous compression algorithm was modified to adapt its quantization level to different ECG signal conditions. Two adaptation strategies were investigated. Both adapt the quantization-step size according to the number of bytes required for storing the coded signal, beat arrival times, and beat classifications. The new compression algorithm was tested with data from the MIT/BIH database on CD ROM. With the more successful of the two strategies, the adaptive compression algorithm stored MIT/BIH records with a difference of only 0.8 bps between the record with the highest data rate and the record with the lowest data rate. The average data rate for the entire database was 193.3 bps. Signal-to-compression noise ratios varied from record to record and varied over time for a given record. Average signal to compression noise ratios varied from 26.82 to 532.83. PMID- 8418968 TI - A simple digital filter to remove line-frequency noise in implantable pulse generators. AB - This study investigated computer simulations and animal studies of a simple digital notch filter to remove 50-and 60-Hz line-frequency-noise interference. The digital notch filter was achieved by computing running subtraction of the intracardiac electrogram and the electrogram recorded one notch-sample period previously. Simultaneous rejection of the two worldwide line frequencies was obtained by computation of the minimum of two separate notch-filter outputs. Power consumption of this numerical algorithm was reduced for applications in implantable devices by operating the notch filters only from 20 msec to 50 msec after a sense event. A sense event was classified as a noise sense if the sum of notch-filter output during this window was less than a preset threshold of the sum of the raw data during the same time window. Otherwise, the sense event was classified as a true sense. The computer simulations determined an optimum threshold value of 33%. The filter was tested in five animal studies using a signal generator to inject additive noise interference. The results indicated that this simple filter could be implemented in an implantable pulse generator and could effectively exclude incorrect line-frequency-noise senses at the notched frequencies. PMID- 8418969 TI - Isolation transformers. PMID- 8418970 TI - Anterior cruciate ligament reconstruction. Autograft or allograft? AB - Refinements in arthroscopic techniques have fostered an upsurge in arthroscopically assisted anterior cruciate ligament reconstruction. This article explores the relative merits of autogenous and allogeneic tissue used for this purpose and describes several important technical points in the author's preferred method of surgery. PMID- 8418971 TI - Extra-articular uses of the arthroscope--acromioclavicular arthroplasty. AB - Arthroscopic resection of the acromioclavicular joint is obviously in its infancy. Nonetheless, the procedure seems to be well founded in theory and laboratory work. The early clinical results seem supportive. PMID- 8418972 TI - Magnetic resonance arthrography of the shoulder. A new technique of shoulder imaging. AB - Although magnetic resonance imaging is very sensitive and even though pathology in the rotator cuff is readily detected, it is often difficult to distinguish between complete rotator cuff tears, partial rotator cuff tears, and area of tendinitis. This article reports the results of a new technique for evaluation of shoulder pathology, which the authors have labeled magnetic resonance arthrography, and compares the results of magnetic resonance arthrography with those of conventional magnetic resonance imaging. PMID- 8418973 TI - Anterior cruciate ligament reconstruction and augmentation with PDS graft. AB - The article briefly explains the biomechanical properties of the PDS and the most important ways of its use in anterior cruciate ligament (ACL) reconstruction. The different techniques of ACL reconstruction with gracilis and semitendinosus tendons are outlined. The authors explain their technique of ACL reconstruction with gracilis and semitendinosus plus PDS-band. The possible variations and pitfalls are pointed out. The results of this surgery are expounded. PMID- 8418974 TI - Prepatellar and olecranon arthroscopic bursectomy. AB - Arthroscopic resection of prepatellar and olecranon bursae is a technically feasible operation. It is not more difficult than removing synovium from the suprapatellar pouch of the knee. Although there have been complications, this procedure appears to avoid the problems about the wound described with the open excisional operations for chronic olecranon and prepatellar bursitis. PMID- 8418975 TI - Arthroscopic microdiscectomy of the lumbar spine. AB - Disc herniation in athletes is uncommon; however, it requires prompt diagnosis and treatment. Arthroscopic microdiscectomy is associated with minimal postoperative morbidity and it is particularly suitable for the surgical treatment of unremitting radiculopathy associated with herniated lumbar disc in athletes. The addition of the arthroscope and the deflecting instruments allows for visualization, identification, and extraction of posterior and posterolateral collagenized nuclear fragments that are believed to be the common cause of nerve root compression and sciatica following disc herniation. PMID- 8418976 TI - Chronic exertional compartment syndrome: correlation of compartment pressure and muscle ischemia utilizing 31P-NMR spectroscopy. AB - In the past 5 years, a great deal of time and effort has been expended in an effort to better define clinical, anatomic, and laboratory parameters of CECS. It is now a well-recognized entity and one that can be readily resolved with fasciotomy. But the reasons for predisposition and the pathophysiologic mechanisms remain obscure. It appears, however, that basing the decisions for fasciotomy on clinical characteristics alone leads to overdiagnosis and excessive surgery. In this series, almost 50% of the referred subjects failed to demonstrate adequate laboratory criterion for the diagnosis of CECS. Fasciotomy in these patients may have effected a cure, but the reasons may be unrelated to increased intracompartmental pressure. Furthermore, in the laboratory diagnosis of CECS, the rate of return to resting compartment pressure following exercise seems more accurate than reliance on resting pressure alone. 31P-NMR has proved valuable in the dynamic assessment of muscle ischemia as reflected by relative PCr concentrations. Finally, although a mechanism explaining the source of pain has not been established by this study, it appears that ischemia is not a significant factor. PMID- 8418977 TI - New concepts of rehabilitation following anterior cruciate reconstruction. AB - Can a knee joint with a torn ACL of 2 years' duration ever be able to return to high performance? Very unlikely indeed. Some realistic expectations follow: 1. The knee joint can never be normal after an ACL reconstruction. 2. Surgery must take place as early after the injury as possible, before secondary joint degeneration takes place. 3. The surgery must employ a tissue that best matches the normal ACL in strength and structure. 4. The surgery must involve as little trauma as possible while restoring knee joint mechanics. 5. Stress, although guarded, must be faced by the knee joint as soon as possible after surgery. 6. Progressive weight bearing starts immediately, combined with quadriceps isometrics. ROM of the knee joint, particularly full extension, is conserved and protected. 7. Progressive active ROM without formal resistance continues for 4 weeks. 8. Progressive formal resistance exercises continue for at least 1 year. 9. Sport-specific tasks commence at 16 weeks, depending on the requirement of the sport and the response of the individual athlete. 10. Recovery will plateau at several stages, with the final plateau at approximately 18 months. Knee instability is an exciting but perplexing problem. Although we have advanced profoundly from the era of Jones, Smiley, and others, we still face many of the same challenges as our predecessors. New technology should not fool us. We are still addressing a major structural failure within the knee joint. Our attempts have been non-surgical and surgical, with repair, reconstruction, and replacement. However, fundamental to all of these hopes has been the reconditioning of the extremity after ACL surgery. Can we do better than our forefathers like Licht and others? No one is certain. This article offers an approach, in some areas our approach, but should not be perceived as a cookbook. Individual responses by our patients, athletes, dictate whether any protocol is too hasty or tardy. It is fundamental that we listen to our patients objectively and analyze the knee as it returns from the surgical aggression. The ultimate success of the rehabilitation process will be based on the marriage of science and realistic expectations. PMID- 8418978 TI - Meniscal allografts. AB - Loss of the meniscus has been proved to be associated with increased joint pressures, mechanical changes, and ultimately hyaline cartilage degradation. Since the first arthritic changes following meniscectomy were appreciated, attempts have been made to alter and reverse the joint deterioration that occurs after removal of the knee fibrocartilage. Replacement of the fibrocartilage with either a prosthetic or biologic implant appears to be the only method of restoring normal joint anatomy. By inserting a meniscus substitute for the removed meniscus, the development of joint pathology should be avoided. This article focuses on the procedure of allogenic implants. Allogenic meniscal implants have been performed in humans for over 8 years. Recent clinical work has shown a rapid increase in the number of implants in the last 3 years with clinical review only now being presented. At present, the orthopedic surgeon has available cryopreserved, fresh-frozen, or frozen and irradiated tissue. Although much work has been performed in the animal with fresh-frozen tissue, the newly appreciated risk of disease transmission may require that all future implants be secondarily sterilized. Regardless of the type of implant, the early results of cell viability studies appear the same. Allogenic implants sustain new cellular ingrowth from the host and the DNA is replaced with host DNA. The ultimate success of this operation is not whether allogenic collagen can be transplanted into a host knee, but whether this tissue can be made to function and to preserve hyaline cartilage. Available data suggest that the technique being used to transplant the meniscus does not preserve normal meniscus function. These menisci may not function as they did in the donor. Additionally, few surgical techniques have been tested mechanically to compare meniscus function after transplantation. For these reasons, although transplant surgery for the meniscus remains an exciting and encouraging procedure to save the knee in a person who has had a total meniscectomy, the operation is currently being limited to those involved in study groups and investigational protocols. The long-term follow-up is at present limited or nonexistent. Objective parameters for evaluating posttransplant meniscus function are only now being collected and reviewed. Meniscal transplantation remains a cautiously optimistic treatment for the future. PMID- 8418979 TI - Arthroscopically assisted proximal quadricepsplasty for patellar instability. AB - Arthroscopy has come to play a major role in the evaluation and treatment of patellofemoral instability. In fact, most patients sustaining a traumatic dislocation or recurrent subluxations are subjected to, at a minimum, an arthroscopic evaluation to assess intraarticular damage and patellar tracking. PMID- 8418980 TI - Impingement in the athlete. AB - There is no doubt that the understanding of shoulder mechanics and pathology will continue to evolve. It is not appropriate to consider that all patients, especially young athletes, who have shoulder pain have "impingement." Rather, a careful evaluation will result in the correct diagnosis and the institution of the appropriate treatment. A step-by-step assessment and management scheme is necessary for each health care professional dealing with athletes. If surgery is deemed necessary, the correction of the anatomic abnormality with minimal disruption of normal anatomy will be the key to allowing the athlete to return to his or her chosen sport. PMID- 8418981 TI - Mechanical stress mechanisms and the cell. An endothelial paradigm. AB - There are important physiological and pathological cardiovascular consequences related to endothelial biomechanical properties. The endothelium, however, is not unique in responding to external forces; virtually all cells accommodate or respond to the mechanical environment. PMID- 8418982 TI - Optical measurements of transmembrane potential changes during electric field stimulation of ventricular cells. AB - We evaluated transmembrane potential changes at the ends of isolated rabbit ventricular myocytes during defibrillation-strength shocks given in the cellular refractory period. The myocytes were stimulated (S1 pulse) to produce an action potential. Then a constant-field shock (S2 pulse) with an electric field of 20 or 40 V/cm was given at an S1-S2 interval of 50 msec. The cells were stained with potentiometric dye (di-4-ANEPPS), and the cell end facing the S2 anode or cathode was illuminated with a laser while the fluorescence was recorded. During S2, the cell end facing the S2 cathode became more positive intracellularly, whereas the cell end facing the S2 anode became more negative intracellularly. The S2-induced transmembrane potential change at the cell end (delta Vm) was determined relative to the amplitude of the S1-induced action potential (APA) in each recording (i.e., delta Vm/APA). In Tyrode's solution containing 4.5 mM potassium, delta Vm/APA for 40-V/cm S2 was 1.36 +/- 0.34 at the cell end facing the S2 cathode and -1.65 +/- 0.61 at the cell end facing the S2 anode (n = 9). For the 20-V/cm S2, delta Vm/APA was 0.61 +/- 0.33 at the cell end facing the S2 cathode and -0.71 +/ 0.33 at the cell end facing the S2 anode (n = 6). The delta Vm/APA was not significantly influenced by 20 mM diacetyl monoxime. These results indicate that large delta Vm values occurred at the ends of the cells during S2. The calculated values of delta Vm, assuming a nominal APA of 130 mV, were 177 and -214 mV for the 40-V/cm S2 and 79 and -93 mV for the 20-V/cm S2. The delta Vm was correlated with cell size (r > or = 0.95) and agreed with values predicted by the S2 electric field strength multiplied by half of the cell length to within 27%. When the potassium concentration was increased to 20 mM, delta Vm/APA for 40 V/cm S2 increased 85% and 67% at the cell ends facing the S2 cathode and anode, respectively (n = 9, p < 0.005 versus 4.5 mM potassium), consistent with reduced APA. Thus, with normal or elevated extracellular potassium, transmembrane potential changes at the ends of cells during defibrillation-type stimulation are large enough to produce activation or recovery of voltage-dependent ion channels and may produce the effects responsible for defibrillation. PMID- 8418983 TI - Abdominal coarctation increases insulin-like growth factor I mRNA levels in rat aorta. AB - We have previously demonstrated specific insulin-like growth factor I (IGF I) mRNA transcripts in cultured endothelial and vascular smooth muscle cells and postulated an important role for IGF I in blood vessel growth responses. The purpose of this study was to characterize IGF I gene expression in a model of aortic coarctation hypertension in the rat. This high-renin model of hypertension is associated with hyperplastic vascular responses. Northern analysis of rat aorta demonstrated four specific IGF I mRNA transcripts sized 7.6, 4.6, 1.8, and 0.9-1.2 kb. Quantitation of aortic IGF I mRNA levels by solution hybridization/RNase protection assay demonstrated induction of IGF I transcripts in the hypertensive aorta; levels more than doubled at 7 days and were still significantly elevated 21 days after coarctation. In situ hybridization analysis indicated that IGF I transcripts were localized primarily to adventitial surfaces in normotensive aorta, with minimal signal detected over vascular cells. In hypertensive aortas, there was an increase in IGF I transcripts primarily over vascular smooth muscle cells. Thus, vascular IGF I gene expression is induced in this model of high-renin hypertension. IGF I may play an important role in autocrine/paracrine-mediated vessel wall remodeling in hypertension. PMID- 8418984 TI - Endothelium-dependent relaxation of hypertensive resistance arteries is not impaired under all conditions. AB - Endothelium-dependent relaxation of mesenteric resistance arteries of spontaneously hypertensive rats (SHRs) and normotensive Wistar-Kyoto (WKY) rats was studied. Acetylcholine-induced relaxation of SHR vessels precontracted with 10 microM norepinephrine was endothelium dependent and attenuated compared with WKY vessels. The impaired response of SHR vessels was normalized by inhibition of cyclooxygenase with indomethacin. Blockade of nitric oxide synthetase with NG nitro L-arginine methyl ester (L-NAME) or inhibition of guanylate cyclase with methylene blue attenuated acetylcholine-induced relaxation of norepinephrine contracted SHR vessels but had no effect on WKY vessels. When vessels were precontracted with 30 nM arginine vasopressin, acetylcholine induced similar degrees of relaxation in both strains. A similar response was detected when lysine vasopressin was used to induce tone. Indomethacin had no effect on relaxation responses of SHR and WKY vessels precontracted with either form of vasopressin. L-NAME and methylene blue partially inhibited acetylcholine-induced relaxation of vasopressin-contracted vessels from both strains. Acetylcholine added at baseline did not induce contraction of vessels from either strain. It is concluded that endothelium-dependent relaxation of SHR resistance arteries is not impaired under all circumstances. Acetylcholine-induced relaxation may be suppressed in SHR resistance arteries when norepinephrine is used to induce contraction as a result of catecholamine-induced production of an endothelium derived contracting factor. Vasopressin, on the other hand, does not elicit production of this contracting factor and may enhance the vasorelaxant action of acetylcholine in resistance arteries of both strains via actions on endothelial or vascular smooth muscle cells. PMID- 8418985 TI - Contraction and relaxation of isolated cardiac myocytes of the frog under varying mechanical loads. AB - The mechanics of cardiac systole and relaxation have been studied primarily at the level of the whole heart or intact muscle. End-systolic pressure-volume relations of frog hearts have been found to be load dependent, whereas those of the mammal are relatively load independent. On the other hand, myocardial relaxation as studied at the muscle level is load independent in the frog but markedly load dependent in the mammal. Interpretation of these studies is complicated because of the unknown contribution of extracellular connective tissue, neurohumoral factors, and, in the case of the heart, the complex chamber geometry. Therefore, it is valuable to study cardiac mechanics at the level of the basic unit of contractile activity--the isolated myocyte. The goal of this study was to subject isolated frog cardiomyocytes to mechanical loading paradigms that mimic those presented to the cells within the heart. In the first part of this study, the afterload and preload of contracting cells were varied to study their effects on the end-systolic force-length relation, which was consistently found to be load independent over the range of isotonic shortening tested (typically 5%). We also investigated the force-length-time response of the cells to test the concept of the heart behaving as a time-varying elastance. Our results suggest that in this regard the frog myocyte behaves like mammalian muscle, and they are consistent with the presence of a small viscosity within the cell. We conclude that the tissue structure of the frog heart may contribute to disparity in mechanical behavior at the different structural levels. In the second part of this study, we subjected isolated frog cardiomyocytes to four different loading paradigms to test the hypothesis that myocardial relaxation in the frog is independent of load. These sequences consisted of afterloaded contractions followed by conventional isotonic-isometric relaxation (ACCR) or afterloaded contractions followed by physiologically reversed isometric-isotonic relaxation (ACPR). Relaxation was measured under isometric conditions using a variable afterload with either the ACCR or ACPR paradigms. The decay of force was independent of the cell length at which it occurred or the amount of shortening prior to it within the contractile cycle. Relaxation also was measured as relengthening of the cell under isotonic late-load conditions, using the ACPR paradigm either with a variable afterload or variable late load. Relengthening had a time course that was unaffected by changes in afterload (i.e., extents of shortening) or late load (equivalent to the filling pressure for the heart).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418986 TI - Alteration of reactivity of native arteries induced by venous graft placement. AB - Occlusion of aortocoronary venous grafts can be due to thrombosis, atherosclerosis, or vasospasm. Investigations have focused on properties of the graft itself, and little is known about the vascular reactivity and function of the native arteries proximal and distal to the vein graft, although spasm of the native artery distal to the graft site has been observed in patients. We hypothesized that the function of the endothelium of the native arteries may be altered after surgery. Autogenous venous grafts were placed in femoral arteries of rabbits to study the reactivity of the native arteries after grafting. Four weeks after graft implantation, the vein graft, ipsilateral vein, and native artery proximal and distal to the graft were removed for in vitro studies. Morphological evaluation by scanning electron microscopy and fluorescence microscopy after labeling with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate indicated the presence of an intact, metabolically active endothelial layer. There was no alteration in the contractile responses to phenylephrine of the arteries, vein grafts, or veins. Precontracted vein grafts, veins, and arterial segments proximal to the grafts relaxed when exposed to endothelium-dependent vasodilators (acetylcholine, arachidonic acid, and substance P), but the native arteries distal to the grafts did not. In bioassay cascade experiments, the distal artery did not release any measurable relaxing factor when exposed to acetylcholine. We conclude that the endothelium of the distal artery did not function normally. The extent and reversibility of altered endothelial function remain to be determined. This observation may help to explain the occurrence of myocardial infarction after aortocoronary bypass grafting in some patients. PMID- 8418987 TI - Age-related changes in sarcoplasmic reticulum Ca(2+)-ATPase and alpha-smooth muscle actin gene expression in aortas of normotensive and spontaneously hypertensive rats. AB - The expression of the sarcoplasmic reticulum (SR) Ca(2+)-ATPase gene and the SR Ca2+ pump function were investigated in thoracic aortas of 5- and 17-week-old normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). The relative level of the two isoforms of SR Ca(2+)-ATPase mRNA expressed in the aorta (i.e., SERCA 2a and SERCA 2b) was determined by quantitative S1 nuclease protection analysis and normalized to the level of alpha-smooth muscle (alpha-Sm) actin mRNA. The level of alpha-Sm actin mRNA itself was normalized to the level of 18S ribosomal RNA using slot-blot hybridization assays. Total SR Ca2+ pump activity was estimated by measuring the rate of oxalate-supported Ca2+ uptake in homogenates. At 5 weeks, the amount of SERCA 2a and SERCA 2b mRNA, normalized to 18S ribosomal RNA, and the ratio of alpha-Sm actin mRNA to 18S RNA were identical in SHR and WKY rats. The Ca2+ pump activity was similar in the two strains of rats at 5 weeks. From 5 to 17 weeks, the amount of SERCA 2a mRNA increased in both strains while the level of SERCA 2b mRNA remained constant. The Ca2+ pump activity was unchanged in SHRs and tended to decrease in WKY rats. Accordingly, the change in the ratio of the SR Ca(2+)-ATPase mRNA isoforms does not appear to influence SR function. The level of alpha-Sm actin mRNA and SERCA 2a mRNA increased in parallel from 5 to 17 weeks in both strains.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8418988 TI - Dystrophin-glycoprotein complex and laminin colocalize to the sarcolemma and transverse tubules of cardiac muscle. AB - The expression and subcellular distribution of the dystrophin-glycoprotein complex and laminin were examined in cardiac muscle by immunoblot and immunofluorescence analysis of rabbit and sheep papillary muscle. The five dystrophin-associated proteins (DAPs), 156-DAG, 59-DAP, 50-DAG, 43-DAG, and 35 DAG, were identified in rabbit ventricular muscle and found to codistribute with dystrophin in both papillary myofibers and Purkinje fibers. The DAPs and dystrophin codistributed not only in the free surface sarcolemma but also in interior regions of the myofibers where T tubules are present. Neither the DAPs nor dystrophin were detected in intercalated discs, a specialized region of cardiac sarcolemma where neighboring myocardial cells are physically joined by cell-cell junctions. Similarly, in bundles of Purkinje fibers, which lack T tubules, DAPs and dystrophin were also found to codistribute at the free surface sarcolemma but were not detected either in the region of surface sarcolemma closely apposed to a neighboring Purkinje fiber or in interior regions of these myofibers. Comparison between the distribution of the dystrophin-glycoprotein complex and laminin showed that laminin codistributes with the components of this complex in both papillary myofibers and Purkinje fibers. These results are consistent with previous findings demonstrating that the extracellularly exposed 156-DAG (dystroglycan) of the skeletal muscle dystrophin-glycoprotein complex binds laminin, a component of the basement membrane. Although we demonstrate that DAPs, dystrophin, and laminin colocalize to the sarcolemma in rabbit and sheep papillary myofibers as they do in skeletal myofibers, the most striking difference between the subcellular distribution of these proteins in cardiac and skeletal muscle is that the dystrophin-glycoprotein complex and laminin also localize to regions of the fibers where T tubules are distributed in cardiac but not in skeletal muscle. These results imply that the protein composition and thus possibly some functions of T tubules in cardiac muscle are distinct from those of skeletal muscle. PMID- 8418989 TI - Reperfusion after brief ischemia disrupts the microtubule network in canine hearts. AB - Histological changes in the stunned myocardium are believed to be minimal. This study examined whether cytoskeletal structures of microtubules are disrupted in the stunned myocardium. In 38 dogs, the left anterior descending coronary artery was occluded for 15 minutes and reperfused to produce the stunned myocardium. Microtubules were stained immunohistochemically. In intact myocardium, microtubules appeared as a filamentous network throughout the cytoplasm and encircled the nucleus. This pattern was not affected by 15 minutes of ischemia. One hour of reperfusion, however, disrupted microtubular structure substantially (disruption score in the endocardium, 53.4 +/- 6.0%) although actin filaments remained intact. Microtubular structures were reconstituted 1-3 days after reperfusion, showing supernormal immunoreactivity. Five days after reperfusion, the pattern of microtubular staining was normal. In another protocol, the role of Ca2+ during reperfusion in microtubular disruption was examined. When intracoronary infusion of EDTA (1.67 mumol/kg body wt per minute) was performed during the initial 10 minutes of reperfusion, myocardial stunning was attenuated. The fractional shortening in the perfused area after 1 hour of reperfusion was 20.1 +/- 1.2% versus 11.5 +/- 0.5% in the control condition (p < 0.05), and the microtubular disruption score was lower (12.6 +/- 1.4%). Although intracoronary infusion of calcium chloride (9 mumol/kg body wt per minute) for 10 minutes in nonischemic hearts increased contractile function (fractional shortening, 25.3 +/ 2.0%), it severely disrupted microtubular networks (microtubular disruption score, 64.0 +/- 10.6%). We conclude that microtubules supporting the structural integrity of myofibrils and other organelles are reversibly disrupted by reperfusion after brief ischemia probably through calcium overload. PMID- 8418990 TI - Modulation of Ca2+ cycling in cardiac myocytes by arachidonic acid. AB - It is believed that inotropic agents exert their effects in cardiac muscle via a modulation of Ca2+ cycling; however, the involvement of phospholipase activation and the biochemical pathways participating in inotropic responsiveness remain unclear. The aim of the current study was to determine whether arachidonic acid and/or eicosanoids participate in inotropic responses by modulating Ca2+ cycling in cardiac myocytes. Experiments were performed with populations of freshly isolated, fura-2-loaded adult rat ventricular myocytes. Arachidonic acid stimulated a transient increase in cytosolic free Ca2+, which was still present after addition of EGTA but was significantly reduced by pretreatment with caffeine. Addition of arachidonic acid to either electrically stimulated or quiescent myocytes enhanced the amplitude of the ATP-induced Ca2+ transient. This effect was still observed in the presence of inhibitors of cyclooxygenase, lipoxygenase, and epoxygenase pathways but was significantly diminished after pretreatment with inhibitors of protein kinase C. In contrast, arachidonic acid attenuated the amplitude of electrically induced Ca2+ transients. This effect was mimicked by eicosatetraynoic acid and by the K+ channel agonist pinacidil. The inhibitory effect of eicosatetraynoic acid and arachidonic acid was reversed by addition of fatty acid-free bovine serum albumin. Together, these results suggest that arachidonic acid may play a physiological role in cardiac muscle excitation contraction coupling as a modulator of sarcolemmal ion channels and/or Ca2+ release from the sarcoplasmic reticulum. PMID- 8418991 TI - Diminished basal nitric oxide release after myocardial ischemia and reperfusion promotes neutrophil adherence to coronary endothelium. AB - We measured changes in basal release of nitric oxide and its effect on polymorphonuclear leukocyte (PMN) adherence to endothelial cells (ECs) in a feline model of myocardial ischemia (90 minutes) and reperfusion. Basal release of nitric oxide from the left anterior descending coronary artery (LAD) after myocardial ischemia/reperfusion and from the control left circumflex coronary artery (LCX) was assessed by NG-nitro L-arginine methyl ester (L-NAME)-induced vasocontraction. L-NAME induced a significant EC-dependent vasocontraction in control LCX rings (0.28 +/- 0.04 g), which was fully reversed by L-arginine but not D-arginine. L-NAME-induced vasocontraction of LAD rings was not significantly changed after 90 minutes of myocardial ischemia without reperfusion. However, 10 minutes of reperfusion reduced the L-NAME-induced vasocontraction to 0.13 +/- 0.04 g (p < 0.05), and this was restored by addition of 3 mM L-arginine but not D arginine. Longer periods of reperfusion progressively decreased L-NAME-induced vasocontraction. After 270 minutes of reperfusion, L-NAME-induced vasocontraction was virtually abolished. Myocardial ischemia without reperfusion did not increase PMN adherence to ECs. However, PMN adherence to LAD ECs was significantly increased after 20 minutes of reperfusion (39 +/- 6 to 105 +/- 9 PMNs/mm2, p < 0.01), and incubation of LAD segments with L-arginine significantly attenuated this increase in PMN adherence. After 270 minutes of reperfusion, PMN adherence to LAD ECs was further increased to 224 +/- 10 PMNs/mm2 (p < 0.001). This increase in PMN adherence was almost completely blocked by MAb R15.7, a monoclonal antibody against CD18 of PMNs, and was significantly attenuated by MAb RR1/1, a monoclonal antibody against intercellular adhesion molecule-1 of ECs (p < 0.01). These results indicate that decreased basal release of endothelium derived relaxing factor after myocardial ischemia/reperfusion precedes enhanced PMN adherence to the coronary endothelium, which may lead to PMN-induced myocardial injury. PMID- 8418992 TI - Effects of thiol protease inhibitors on cell cycle and proliferation of vascular smooth muscle cells in culture. AB - Smooth muscle proliferation is a prominent feature of the vascular response to mechanical injury. Accordingly, modulation of proliferation has important therapeutic implications for angioplasty restenosis. We have identified a subclass of thiol protease inhibitors (TPIs) that reversibly inhibit bovine aortic smooth muscle cell (BASMC) proliferation in vitro. To define the nature of this inhibition, an evaluation of selected steps in the cell cycle was undertaken. Treatment of BASMCs with benzyloxycarbonyl-Leu-norleucinal (calpeptin) at 100 microM and acetyl-Leu-Leu-norleucinal (TPI-1) at 50 microM was shown to cause a block of platelet-derived growth factor-BB as well as serum inducible cell cycle progression at a point before the G1-S boundary, reducing the percentage of bromodeoxyuridine-positive cells from 87% to 5% over a 24-hour labeling period. Addition of TPI-1 at various times after serum addition to serum deprived BASMCs showed 80% of the maximal block of DNA synthesis even when added 6 hours after serum. The cell cycle progression block was gradually lost as the delay from serum to TPI-1 application was increased from 6 to 12 hours. By Northern analysis of mRNA after serum addition, TPI-1 caused a fourfold decrease in the transient elevation of fos and myc proto-oncogene as well as a decrease in the levels of both muscle and nonmuscle actin mRNA induced early after serum addition. Flow cytometric analysis of DNA content and synthesis in BASMCs treated with TPI-1 or calpeptin additionally revealed the presence of a distinct cell cycle block in the G2-M compartment. In the aggregate, these results suggest the existence of more than one molecular site potentially involved in inhibition by TPI of cell cycling in BASMCs. PMID- 8418993 TI - Respiratory modulation of muscle sympathetic nerve activity in intact and lung denervated humans. AB - We determined the influences of breathing-induced changes in intrathoracic and intravascular pressures, central respiratory drive, and pulmonary vagal feedback on the within-breath variation in skeletal muscle sympathetic nerve activity (MSNA) in humans. MSNA (peroneal microneurography), arterial blood pressure (Finapres finger monitor), and tidal volume (VT) were recorded continuously in six normal subjects and four heart-lung transplant patients during: 1) spontaneous air breathing; 2) increased FICO2; 3) voluntary augmentation of VT with and without inspiratory resistance; and 4) positive pressure, passive mechanical ventilation. During conditions 3 and 4, which were performed under isocapnic conditions with a high MSNA background (either high resting activity or nonhypotensive lower body suction), subjects breathed at control or elevated VT with normal or prolonged inspiratory time (TI); breathing frequency was 12 breaths per minute. During control breathing in normal subjects there was a distinct within-breath pattern of MSNA, with approximately 70% of the activity occurring during low lung volumes (initial half of inspiration and latter half of expiration). This within-breath variation of MSNA was potentiated with increased VT breathing (> 85% of activity occurring during low lung volumes; p < 0.05 versus control breathing) and was similar during the voluntary and CO2-induced hyperpneas. MSNA decreased progressively and markedly from onset to late inspiration; fell slightly further, reaching its nadir at end-inspiration/onset expiration; and rose sharply during mid-late expiration. Only the nadir of MSNA was associated with any change in arterial pressure. Resistive breathing, especially at elevated VT, caused a fall in arterial pressure and increased respiratory drive during inspiration, yet MSNA still declined as lung volume increased. Normal within-breath modulation of MSNA also was observed during control and elevated VT induced via positive pressure with passive ventilation, which reversed lung inflation/deflation-induced intrathoracic pressure changes and reduced or removed respiratory motor output. During control breathing in transplant patients the specific within-breath pattern of MSNA was somewhat different than that of the normal subjects, but on average, the overall low lung volume to high lung volume MSNA ratio was similar to normal subjects. In contrast to the normal subjects, however, there was no potentiation of the within-breath variation of MSNA with elevated tidal breathing. These findings indicate that during normal levels of tidal breathing most of the respiratory phase influence on muscle sympathetic outflow observed in normal conscious humans is independent of baroreceptor-sensed fluctuations in intrathoracic or intravascular pressures and of lung inflation-stimulated vagal afferent activity.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8418994 TI - Residual strain in the ventricle of the stage 16-24 chick embryo. AB - Residual stress and strain, i.e., the stress and strain remaining in a solid when all external loads are removed, may be produced in biological tissues by differential growth. During cardiac development, residual stress and strain may play a role in cardiac morphogenesis by affecting ventricular wall stress. After a transmural radial cut, a passive ventricular cross section opens into a sector, and the size of the opening angle provides a measure of the circumferential residual strain. Residual strains were characterized in this manner for the apical region of the diastolic embryonic chick heart for Hamburger-Hamilton stages 16, 18, 21, and 24 (approximately 2.5, 3.5, 4.0, and 4.5 days, respectively, of a 21-day incubation period). The average opening angle at these stages was 107 +/- 10 degrees, 79 +/- 10 degrees, 73 +/- 11 degrees, and 74 +/- 7 degrees, respectively (n > or = 5 for each stage). These measured angles were correlated with changes in ventricular morphology. Scanning electron micrographs of the apex revealed that the wall of the ventricle is smooth at stage 16. Then at stage 18, myocardial trabeculae develop, forming ridges with primarily a circumferential orientation. By stage 21, the trabeculae develop into a mesh, giving the ventricular wall a spongelike appearance, and the preferred orientation is lost by stage 24. The large decrease in opening angle between stages 16 and 18 corresponded to the onset of trabeculation, which is the greatest change in form during the studied stages. We speculate that residual strain is an important biomechanical factor during cardiac morphogenesis. PMID- 8418995 TI - Alterations in sarcoplasmic reticulum gene expression in human heart failure. A possible mechanism for alterations in systolic and diastolic properties of the failing myocardium. AB - Recent studies have shown that intracellular Ca2+ handling is abnormal in the myocardium of patients with end-stage heart failure. Muscles from the failing hearts showed a prolonged Ca2+ transient and a diminished capacity to restore a low resting Ca2+ level during diastole. Accordingly, we examined whether this defect in Ca2+ transport function is due to alterations in sarcoplasmic reticulum gene expression. We determined the messenger RNA (mRNA) levels of sarcoplasmic reticulum Ca2+ transport proteins in failing human hearts from 17 cardiac transplant recipients with a diagnosis of dilated cardiomyopathy, primary pulmonary hypertension, or ischemic heart disease. The expression levels of each mRNA were compared with each other and then correlated with that of atrial natriuretic factor (ANF) mRNA in the failing ventricle. The mRNA levels for the calcium release channel (ryanodine receptor, RYR2), Ca2+ uptake pump (Ca(2+) ATPase, SERCA2 isoform), and phospholamban differed significantly between heart samples but showed an inverse relation with that of ventricular ANF mRNA. In contrast, calsequestrin mRNA levels remained unchanged in these failing hearts. In addition, beta-myosin and alpha-cardiac actin mRNA levels also showed an inverse relation with ANF mRNA levels. These changes were observed in both right and left ventricles of hearts with congestive heart failure due to dilated cardiomyopathy, primary pulmonary hypertension, or ischemic heart disease. The results are consistent with the hypothesis that abnormal calcium handling in the sarcoplasmic reticulum of failing hearts is due to the altered expression of the genes encoding sarcoplasmic reticulum proteins. PMID- 8418996 TI - Current diagnostic techniques of assessing myocardial viability in patients with hibernating and stunned myocardium. PMID- 8418997 TI - Critical analysis of the signal-averaged electrocardiogram. Improved identification of late potentials. AB - BACKGROUND: This study performed a critical analysis of signal-averaging methods. The objective was to optimize detection of late potentials. METHODS AND RESULTS: We studied two patient populations: a low-arrhythmia-risk group with no evidence of heart disease and a group with clinically documented ventricular tachycardia (VT). Filtered QRS duration (QRSD) and terminal QRS amplitude (RMS40) were measured from the vector magnitude. A QRS duration based on the latest detectable ventricular activity in any of the three individual XYZ leads was also measured. Because of improved signal-to-noise ratio, both individual lead analysis and extended (600-versus 200-beat) averaging yielded significant changes in signal averaged ECG parameters. Both approaches gave an increased sensitivity for VT identification. Sensitivity, specificity, and accuracy were evaluated as functions of critical values of QRSD and RMS40. RMS measurements in the terminal QRS, ranging from 20 to 100 msec and including RMS40, did not contribute to maximizing sensitivity and were highly correlated with QRSD. Our results from the low-arrhythmia-risk group suggest that age and sex should be considered in the definition of late potentials. CONCLUSIONS: We propose a VT risk stratification scheme using signal-averaged ECG parameters obtained from both individual lead and vector magnitude analysis. This allows definition of four categories of VT risk derived statistically from the study data. This definition is based on combined measures of sensitivity, specificity, and negative and positive predictive value. PMID- 8418998 TI - Use-dependent prolongation of ventricular tachycardia cycle length by type I antiarrhythmic drugs in humans. AB - BACKGROUND: Type I antiarrhythmic drugs block the cardiac sodium channel in a use dependent fashion. This use-dependent behavior causes increased drug binding and consequently increased sodium channel blockade at faster stimulation rates. Importantly, the kinetics of drug association and dissociation from the sodium channel differ for each type I antiarrhythmic drug. METHODS AND RESULTS: Thirty five patients receiving type I antiarrhythmic drugs for the treatment of sustained monomorphic ventricular tachycardia (VT) were studied before and after drug therapy. A total of 41 drug studies were performed (lidocaine, n = 10; procainamide, n = 16; flecainide, n = 15). Sustained monomorphic VT of an identical electrocardiographic morphology was induced during the control and follow-up drug studies. During the control study, there was no significant change in the VT cycle length over time. Compared with control, significant prolongation of the onset VT cycle length was observed after treatment with procainamide and flecainide (increase of 52 +/- 24 and 80 +/- 49 msec, respectively) but not after treatment with lidocaine (increase of 8 +/- 37 msec). Additional drug-induced prolongation of the VT cycle length occurred during a 40-second observation period. This secondary "use-dependent" cycle length prolongation contributed significantly to the steady-state VT cycle length during treatment with flecainide (increase of 82 +/- 34 msec; p < 0.0001). Although a use-dependent increase in VT cycle length was observed with procainamide and lidocaine, the increase was not statistically significant (increase of 12 +/- 15 and 8 +/- 8 msec, respectively). The estimated time constants for the onset of use-dependent VT cycle length prolongation were distinctly different for the three drugs. Flecainide's prolongation of the VT cycle length occurred slowly, with an estimated time constant of 12.5 +/- 5.0 seconds. In contrast, the time course of VT cycle length prolongation was rapid during treatment with lidocaine and intermediate during treatment with procainamide (time constants of 0.52 +/- 0.51 and 4.0 +/- 1.3 seconds, respectively). CONCLUSIONS: Use-dependent prolongation of VT cycle length during treatment with type I antiarrhythmic drugs was observed in humans. This effect was clinically significant during treatment with flecainide (i.e., the use-dependent slowing of the heart rate improved the hemodynamic tolerance of the arrhythmia). Finally, the estimated time constants for the use-dependent prolongation of VT cycle length by the three test drugs are similar to their reported in vitro time constants for use-dependent sodium channel blockade. PMID- 8418999 TI - Adenosine-sensitive ventricular tachycardia. Clinical characteristics and response to catheter ablation. AB - BACKGROUND: Sustained ventricular tachycardia in the absence of structural heart disease may have diverse mechanisms. Termination of the tachycardia by adenosine suggests triggered automaticity as the etiology in many of these patients. We examined the clinical characteristics, electrophysiological responses, and results of catheter ablation in this patient subgroup. METHODS AND RESULTS: Intravenous adenosine terminated sustained ventricular tachycardia in seven of 14 consecutive patients without evidence of structural heart disease. In each of these patients, the tachycardia had a left bundle branch block, inferior-axis QRS configuration and occurred predominantly during stress or exertion. A morphologically similar sustained tachycardia was induced in six of seven patients during programmed ventricular stimulation, although day-to-day reproducibility was poor. Signal-averaged ECGs were normal in all patients. Imaging with 123I-metaiodobenzylguanidine did not reveal focal abnormalities in any of five patients. A discrete site of origin was identified in the free wall of the pulmonary infundibulum in all patients. Limited application of direct current shocks (two patients) or radiofrequency energy (five patients) resulted in long-term abolition of spontaneous and inducible ventricular tachycardia in all patients. CONCLUSIONS: Adenosine-sensitive ventricular tachycardia appears to arise from relatively discrete sites predominantly located in the free wall of the pulmonary infundibulum. The localized nature of this tachycardia renders it amenable to long-term cure by catheter ablation techniques. PMID- 8419000 TI - Pace mapping using body surface potential maps to guide catheter ablation of accessory pathways in patients with Wolff-Parkinson-White syndrome. AB - BACKGROUND. A pace mapping technique using body surface potential maps (BSPMs) was developed to guide the positioning of an ablation catheter at the ventricular insertion point of accessory pathways (AP) in patients with the Wolff-Parkinson White syndrome (WPW). METHODS AND RESULTS. The study was performed on 30 WPW patients. BSPMs were recorded with 63 leads distributed over the entire torso surface. The catheter used for radiofrequency ablation was first placed in the vicinity of the ventricular preexcitation site predicted by BSPMs recorded during the delta wave. BSPMs were then recorded during pacing with this catheter, the comparison between the preexcited and paced BSPMs indicated whether the pacing site was too anterior or posterior with respect to the preexcitation site, and the catheter was moved accordingly. This process was repeated until the preexcited and paced BSPMs were highly correlated (r > or = 0.8), and ablation then was attempted. It was possible to successfully ablate the AP in 28 patients after an investigation that lasted 54 +/- 44 minutes between the recording of the first paced BSPM and that of the BSPM paced at the successful ablation site. Patients with left free wall pathways needed less investigation time compared with patients with pathways of other locations (46 +/- 9 versus 100 +/- 25 minutes, p = 0.031). The sensitivity of BSPM pace mapping was assessed using pacing with a multipolar catheter, and significant changes were observed on the BSPMs for beats with pacing sites that were only 5 mm apart. CONCLUSIONS, BSPM pace mapping allowed us to achieve a 93% success rate with short investigation durations, provides significant information that cannot be obtained with the standard 12-lead ECG, is a self-correcting procedure that reduces the importance of BSPM alterations due to individual differences in the shape of the torso or heart, and is applicable only to patients with AP showing antegrade conduction. PMID- 8419001 TI - Beneficial effects of atrial natriuretic peptide on exercise-induced myocardial ischemia in patients with stable effort angina pectoris. AB - BACKGROUND: It has been shown that atrial natriuretic peptide (ANP), an endogenous vasodilator, dilates coronary arteries and decreases coronary vascular resistance. The purpose of this study was to determine whether an intravenous administration of ANP attenuated exercise-induced myocardial ischemia in 14 patients with stable effort angina pectoris. METHODS AND RESULTS: The first 12 patients (patients 1-12) who had exercise-induced ST segment depression underwent treadmill exercise testing and the last seven patients (patients 8-14) underwent the exercise 201Tl-single-photon emission computed tomography (SPECT) study while synthetic 28-amino acid alpha-human ANP (0.1 micrograms/kg per minute) or saline was intravenously infused in a double-blind, cross-over manner. The duration of exercise testing was the same during ANP and saline infusion, which was determined in preliminary exercise testings in each patient to cause a transient perfusion defect and/or ischemic ST segment depression. During saline infusion, all 12 patients developed exercise-induced ischemic ST segment depression, whereas no significant ST segment depression appeared during ANP infusion. Average ST segment depression during ANP infusion was significantly less (p < 0.01) than that during saline infusion (0.0 +/- 0.0 versus 0.2 +/- 0.1 mV, mean +/- SD). The averaged extent and severity scores assessed by 201Tl-SPECT were smaller (p < 0.05) during ANP infusion than during saline infusion (extent score: 0.22 +/- 0.20 versus 0.42 +/- 0.20; severity score: 18.77 +/- 23.45 versus 38.24 +/- 24.04, respectively). ANP decreased resting systolic blood pressure from 125 +/- 15 to 110 +/- 15 mm Hg (p < 0.01) but did not alter resting heart rate. At peak exercise, systolic blood pressure, heart rate, and the rate-pressure products did not differ during ANP and saline infusion. At peak exercise, plasma ANP increased from 98 +/- 45 to 4,383 +/- 2,782 pg/ml and cGMP increased from 3.6 +/- 1.7 to 34.5 +/- 16.1 pmol/ml during ANP infusion; values were significantly higher than those during saline infusion (from 96 +/- 42 to 133 +/- 66 pg/ml and from 3.4 +/- 1.8 to 4.6 +/- 1.8 pmol/ml, respectively). CONCLUSIONS: An intravenous administration of ANP attenuated exercise-induced myocardial ischemia in patients with stable effort angina pectoris. Although the mechanism by which ANP attenuated myocardial ischemia was not defined, increased myocardial perfusion to the ischemic region might be an important factor. PMID- 8419002 TI - Association of early-onset coronary heart disease in South Asian men with glucose intolerance and hyperinsulinemia. AB - BACKGROUND: Rates of coronary heart disease are higher in South Asians (Indians, Pakistanis, and Bangladeshis) settled overseas than in other ethnic groups. We tested the hypothesis that this excess risk results from metabolic disturbances associated with insulin resistance. METHODS AND RESULTS: There were 1,421 South Asian men and 1,515 European men aged 40-69 years in London examined in the Southall Study. Prevalence of ischemic ECG abnormalities was higher in South Asians than in Europeans (17% versus 12%, p < 0.001), with an excess of major Q waves (Minnesota codes 1-1 or 1-2) in younger South Asian men (p = 0.01 for the age-ethnicity interaction). Major Q waves were strongly associated with glucose intolerance and hyperinsulinemia in younger South Asians; these factors accounted statistically for 73% of major Q waves in those aged 40-54 years. Age standardized prevalence of a positive history of coronary heart disease was similar in South Asians and Europeans (8.5% versus 8.2%, NS), and positive history without Q waves was not associated with glucose intolerance or hyperinsulinemia in South Asians. Smoking rates and average plasma cholesterol were lower in South Asians than in Europeans; in a logistic model controlling for smoking and cholesterol, the odds ratio for major Q waves in South Asians compared with Europeans was 2.4 (95% CI, 1.5-3.8). Adjusting for glucose intolerance and hyperinsulinemia reduced this ratio to 1.5 (95% CI, 0.9-2.5). CONCLUSIONS: These results are consistent with the hypothesis that insulin resistance underlies the high coronary risk in South Asian people and strengthen the evidence for a fundamental role of this metabolic pattern in the etiology of coronary heart disease. PMID- 8419003 TI - Indobufen in the prevention of thromboembolic complications in patients with heart disease. A randomized, placebo-controlled, double-blind study. AB - BACKGROUND: The purpose of this randomized, double-blind study was to evaluate the efficacy of indobufen, a reversible inhibitor of platelet cyclooxygenase, in the prevention of embolic events of cardiac origin. METHODS AND RESULTS: One hundred ninety-six patients with heart disease and at risk for cardiogenic embolism (90 with atrial fibrillation and 106 in sinus rhythm) were randomly assigned to receive indobufen (100 mg b.i.d.) or placebo. All patients were reexamined every 3 months for the duration of the study. The primary study end points were cerebral ischemic attack (stroke and transient ischemic attack), systemic embolism, pulmonary embolism, and fatal myocardial infarction. The median duration of treatment was 854 days in the indobufen group and 865 days in the placebo group. The frequencies of primary end points (fatal and nonfatal) were 6.1% and 17.3%, respectively, in the indobufen and placebo groups (p < 0.05) for a reduction of 65% in the risk of a primary event (indobufen/placebo relative risk, 0.35; 95% confidence limits, 0.14-0.89). Adverse drug reactions, mostly gastrointestinal or hemostasis disorders, occurred in 9.2% of indobufen-treated patients. CONCLUSIONS: The results of the study indicate that indobufen may reduce the risk of ischemic events in patients with heart disease associated with an increased risk of embolism. PMID- 8419004 TI - Accuracy of exercise 201Tl myocardial scintigraphy in asymptomatic young men. AB - BACKGROUND: Little is known about the diagnostic usefulness of 201Tl scintigraphy for detecting asymptomatic coronary artery disease in apparently healthy men. We thus evaluated planar 201Tl exercise myocardial scintigraphy in 845 asymptomatic male military aircrew undergoing coronary arteriography because of abnormal noninvasive tests suggesting possible myocardial ischemia. METHODS AND RESULTS: Patients were stratified by prior disease risk into six subgroups using age (< 45 and > or = 45 years) and ratio of total to high density lipoprotein cholesterol (< 4.5, 4.5-6.0, and > 6.0). Significant coronary artery disease (> or = 50% diameter stenosis in any major coronary artery) was present in 143 (16.9% prevalence). Overall sensitivity and specificity of 201Tl scintigraphy adjusted for verification bias were estimated to be 45 +/- 4% and 78 +/- 1%, respectively. These values are lower than corresponding values accepted for clinical populations. Positive and negative predictive values varied across subgroups. A normal thallium scan indicated low risk of disease, but an abnormal test was likely to be a false-positive result. A logistic equation was retrospectively fit to the data for estimating the probability of disease given age, cholesterol ratio, and thallium results. Within each quintile of estimated risk, the average risk did not differ significantly from the observed disease prevalence. CONCLUSIONS: Exercise 201Tl scintigraphy is limited by the frequent occurrence of false-positive tests in detecting asymptomatic, anatomic coronary artery disease in young men in accordance with Bayesian probability theory. PMID- 8419005 TI - Quantification of regional myocardial perfusion with generator-produced 62Cu-PTSM and positron emission tomography. AB - BACKGROUND: Noninvasive assessment of regional myocardial perfusion at rest and after stress is important for the objective evaluation of the effects of coronary artery disease and its response to therapy. Centers that do not have cyclotrons rely on generator-produced radioisotopes for assessment of regional myocardial perfusion with positron emission tomography (PET). The aim of the present study was to develop and implement an approach to quantify regional myocardial perfusion using copper(II) pyruvaldehyde bis-(N4-thiosemicarbazone) (PTSM) labeled with the generator-produced, positron-emitting radionuclide 62Cu (t1/2 = 9.7 minutes). METHODS AND RESULTS: Regional perfusion was estimated from dynamic PET scans after intravenous administration of 62Cu-PTSM in 21 studies in 13 intact dogs evaluated over a wide range of myocardial flow values. In 15 interventions in nine dogs, regional perfusion was also estimated with H2(15)O. Regional perfusion with 62Cu-PTSM was estimated from dynamic blood and tissue time-activity curves, along with the model parameter k1 (forward rate of transport) and the PET parameter FBM (fraction of blood pool activity observed in tissue), using a two-compartment kinetic model. Arterial blood activity was corrected for red blood cell-associated 62Cu. In 44 comparisons, estimates of regional perfusion with 62Cu-PTSM correlated well with estimates obtained with concomitantly administered radiolabeled microspheres (y = 0.90x +/- 0.15, r = 0.95, p < 0.05) over a flow range from 0.23 to 6.14 ml/g per minute. In five healthy human volunteers evaluated at rest with H2(15)O and 62Cu-PTSM, regional perfusion estimated with 62Cu-PTSM was not significantly different from that obtained with H2(15)O (1.05 +/- 0.36 versus 0.96 +/- 0.28 ml/g per minute). 62Cu PTSM provided high-quality images of the heart. CONCLUSIONS: The results of this study demonstrate that quantification of regional myocardial perfusion is feasible using generator-produced 62Cu-PTSM. Since 62Cu-PTSM can be used to estimate perfusion in the brain, kidney, and tumors as well as in the heart, it is an attractive tracer for centers that rely on generator-produced tracers for the evaluation of perfusion with PET. PMID- 8419007 TI - Angiotensin II directly stimulates release of atrial natriuretic factor in isolated rabbit hearts. AB - BACKGROUND: Previous studies have shown that infusion of angiotensin II (Ang II) increases plasma concentrations of atrial natriuretic factor (ANF) in vivo. This phenomenon has been considered secondary to the effects of Ang II on cardiac and systemic hemodynamics. The present study was designed to assess whether Ang II may exert a direct stimulatory effect on ANF release from the heart independent of changes in hemodynamics. METHODS AND RESULTS: Isolated rabbit hearts were perfused in the Langendorff mode. Heart rate, coronary flow, and atrial and left ventricular (LV) volumes were kept constant. After stabilization, Ang II was infused intracoronary at increasing doses (10(-11) to 10(-8) M) in nine hearts and at a single dose of 10(-10) M in 10 hearts. Each infusion lasted for 5 minutes and was followed by a 10-minute washout period. Four hearts received vehicle alone for 80 minutes. Ang II induced a dose-dependent increase in coronary perfusion pressure and in LV developed pressure. ANF release, measured by radioimmunoassay on the extracts of the cardiac effluent, also increased during Ang II infusion and returned to the basal values during the 10-minute washout period. In the control group, coronary perfusion pressure, LV developed pressure, and LV end-diastolic pressure did not change appreciably over the observation period, whereas ANF release progressively decreased during perfusion. CONCLUSIONS: Ang II can directly stimulate cardiac release of ANF in isolated rabbit hearts independently of changes in hemodynamics. PMID- 8419006 TI - 8-Methoxypsoralen and longwave ultraviolet irradiation are a novel antiproliferative combination for vascular smooth muscle. AB - BACKGROUND: Smooth muscle cell proliferation plays a major role in the genesis of restenosis after angioplasty or vascular injury. Although the effects of arterial exposure to high-energy radiation sources such as laser have been investigated in detail, the effects on vascular cells of low-intensity radiant energy in combination with photoactive agents have not been extensively characterized. Psoralens are photoactive agents that are known to be well tolerated when used in conjunction with local exposure to ultraviolet light in the A band (UVA) for the treatment of various dermatologic proliferative disorders. METHODS AND RESULTS: We have investigated the effects of psoralen/UVA (PUVA) exposure on the proliferation of bovine aortic smooth muscle cells. Proliferation and viability were assessed over a 14-day period by trypan blue exclusion counts. Cell cycle effects were evaluated by thymidine incorporation and flow cytometry with DNA quantitation after addition of serum or platelet-derived growth factor B-chain (PDGF-BB) to subconfluent cells synchronized by serum withdrawal. No effect was observed after exposure to 8-methoxypsoralen (8-MOP) at concentrations up to 10 microM or UVA irradiation at energies up to 2.5 J/cm2. Longwave ultraviolet light and 8-MOP were found to behave synergistically as potent inhibitors of DNA synthesis in bovine aortic smooth muscle cells with the EC50 in combination ranging from 7 microM at 0.35 J/cm2 to 0.2 microM at 2.1 J/cm2. Similar antiproliferative effects were obtained by an inverse variation of dose and energy delivered. After serum stimulation, inhibition of DNA synthesis was found with either an immediate or delayed (16-hour) application of PUVA. This effect was independent of subsequent 8-MOP washout. Flow cytometry of cells treated with PUVA at several times after serum stimulation demonstrated for each time point a block in further cell cycle progression for cells in all phases of the cell cycle. Evaluation of [125I]-labeled PDGF and epidermal growth factor (EGF) binding revealed no effect of PUVA on the apparent number or affinity of PDGF binding sites present but did reveal a dose-dependent inhibition by PUVA of EGF binding. This inhibition of EGF binding occurred increasingly at higher PUVA doses than the cell cycle inhibition and accordingly did not appear to represent a critical mechanism for the antiproliferative effect. Cell counting after a single exposure to PUVA (1 microM, 1.5 J/cm2) revealed complete stasis of cell proliferation over a 28-day period without recurrent exposure. No increase in trypan-positive cells was noted over this period. CONCLUSIONS: PUVA treatment represents a novel method for locally inhibiting proliferation of vascular smooth muscle cells without producing cytolysis. PMID- 8419008 TI - Myocardial electrical impedance mapping of ischemic sheep hearts and healing aneurysms. AB - BACKGROUND: This study was designed to examine the bulk electrical properties of myocardium and their variation with the evolution of infarction after coronary occlusion. These properties may be useful in distinguishing between normal, ischemic, and infarcted tissue on the basis of electrophysiological parameters. METHODS AND RESULTS: The electrical impedance of myocardial tissue was studied in a sheep model of infarction. The animal model involved a one-stage ligation of the left anterior descending and second diagonal arteries at a point 40% of the distance from the apex to the base. By use of a four-electrode probe, an epicardial mapping system was developed that allowed for cardiac cycle gated and signal-averaged measurements. Subthreshold current (15 microA) was injected through two of the electrodes at frequencies of 1, 5, and 15 kHz and the induced potential measured with the other two electrodes. Epicardial maps of the left ventricle were obtained during acute infarction and at 1-, 2-, and 6-week intervals after occlusion. Results showed the average specific impedance of the myocardium before infarction to be 158 +/- 26 omega-cm independent of location on the epicardium. By 60 minutes after coronary occlusion, the specific impedance had increased by 199% (p < 0.005, n = 9); it remained elevated for up to 4 hours. One week after infarction, the specific impedance decreased to 59% of the control value (p < 0.025, n = 8). Six weeks after occlusion, the specific impedance remained low at 57% of that of the noninfarcted tissue (p < 0.005, n = 9). The phase angle of the complex impedance was also measured and revealed similar changes. The hydroxyproline content of the tissue was assayed to assess infarct healing. CONCLUSIONS: In this animal model, impedance is a bulk electrical property of tissue that varies with the evolution of myocardial infarction. Impedance mapping revealed significantly different values for normal, ischemic, and infarcted tissue and may prove useful in better defining the electrophysiological characteristics of such tissue. PMID- 8419009 TI - Ischemic preconditioning increases adenosine release and 5'-nucleotidase activity during myocardial ischemia and reperfusion in dogs. Implications for myocardial salvage. AB - BACKGROUND: Adenosine has been reported to mediate the necrosis-limiting effects of ischemic preconditioning; however, it is unclear how this mediation occurs. The present study was undertaken to test the hypothesis that ischemic preconditioning increases 5'-nucleotidase activity and adenosine release during sustained ischemia and subsequent reperfusion. METHODS AND RESULTS: After thoracotomy, the left anterior descending coronary artery was cannulated and perfused with blood redirected from the left carotid artery in 32 dogs. Ischemic preconditioning was produced by four cycles in which the coronary artery was occluded and then reperfused for 5 minutes each. After the last cycle of ischemia and reperfusion, the coronary artery was occluded for 40 minutes. This was followed by 120 minutes of reperfusion. In the control group, the coronary artery was occluded for 40 minutes and reperfused for 120 minutes without ischemic preconditioning. The plasma adenosine concentration was measured and blood gases were analyzed in coronary arterial and venous blood samples taken during 120 minutes of reperfusion. Myocardial 5'-nucleotidase activity was measured before and at 40 minutes of sustained ischemia with and without ischemic preconditioning. The adenosine concentration in coronary venous blood during reperfusion was significantly higher in preconditioned hearts than in the control hearts: 1 minute after the onset of reperfusion, 546 +/- 57 versus 244 +/- 41 pmol/ml; 10 minutes after, 308 +/- 30 versus 114 +/- 14 pmol/ml; 30 minutes after, 175 +/- 24 versus 82 +/- 16 pmol/ml, respectively (p < 0.01). Ectosolic and cytosolic 5'-nucleotidase activities increased in both endocardial and epicardial myocardium in the ischemia-preconditioned hearts. Furthermore, 40 minutes of ischemia increased 5'-nucleotidase activity in ischemia-preconditioned hearts more than in control hearts. CONCLUSIONS: Ischemic preconditioning increases adenosine release and 5'-nucleotidase activity during sustained ischemia and subsequent reperfusion. PMID- 8419010 TI - Long-term outcome of patients with depressed left ventricular function undergoing percutaneous transluminal coronary angioplasty. The NHLBI PTCA Registry. AB - BACKGROUND: Coronary revascularization with bypass has been shown to improve survival in patients with coronary artery disease and left ventricular dysfunction. In these patients, use of nonsurgical revascularization with percutaneous transluminal coronary angioplasty (PTCA) is increasing, although their long-term outcome has not been well delineated. The purpose of this investigation was to characterize the outcome of angioplasty in patients with decreased left ventricular function and contrast it with the results in patients with normal left ventricular function. METHODS AND RESULTS: In the 1985-1986 National Heart, Lung, and Blood Institute's PTCA Registry, of 1,802 patients undergoing PTCA, 244 patients (13.5%) had an ejection fraction of < or = 45% (mean, 39.6 +/- 6.8%). These patients had a higher incidence of prior infarction, a longer and worse history of manifestations of coronary disease, and more extensive coronary artery disease than patients with well-preserved function; 88% and 91%, respectively, had successful dilation of at least one lesion (nonsignificant difference). However, patients with decreased left ventricular function had a decreased frequency of successful dilation of all lesions in which PTCA was attempted (76% versus 84%, p < 0.01). There were no statistically significant differences in in-hospital complications--death occurred in 0.8% and 0.7%, nonfatal myocardial infarction occurred in 4.9% and 4.5%, and emergency surgical revascularization was performed in 4.5% and 3.2%, respectively. Patients were followed for a mean of 4.1 years; during this time, patients with decreased left ventricular function had significantly worse survival and combined event free survival. Despite this, at 4 years, 87% of the patients with a mean ejection fraction of 39.6% remained alive, and 77% were alive and had not experienced infarction or required bypass. CONCLUSIONS: PTCA is effective in selected patients with depressed left ventricular function. Initial outcome and risk benefit ratio are excellent. Successful dilation of at least one vessel was achieved in 88% of patients with depressed left ventricular function and in 91% of patients with more normal left ventricular function. The former group, however, had a decreased incidence of successful dilation in all lesions in which dilation was attempted (76% versus 84%, p < 0.01). There was no significant difference in in-hospital complications between the two groups. During follow-up, patients with decreased left ventricular function had worse event-free survival, although 77% were alive without infarction or bypass grafting at 4 years. PMID- 8419011 TI - Effects of subendocardial ablation on anodal supernormal excitation and ventricular vulnerability in open-chest dogs. AB - BACKGROUND: In Langendorff-perfused hearts and in hearts on cardiopulmonary bypass, chemical ablation of the subendocardium of both ventricles decreases ventricular vulnerability to fibrillation. It was hypothesized that the effects of ablation are a result of the elimination of the subendocardial Purkinje fiber network. This hypothesis has been supported by recent observations that the supernormal excitability that is demonstrable in the Purkinje fibers is associated with arrhythmogenesis. METHODS AND RESULTS: We tested this hypothesis on 10 open-chest dogs by evaluating the strength-interval curves of anodal and cathodal stimulation with the assistance of computerized mapping techniques. The ventricular fibrillation threshold was also determined. The same test was then performed after chemical ablation of the subendocardium of either the right ventricle (six dogs) or both ventricles (four dogs). Anodal supernormality was consistently demonstrated in all the dogs studied both before and after subendocardial ablation. The ventricular fibrillation thresholds were 23 +/- 5 mA both before and after right ventricular subendocardial ablation (p = NS). The ventricular fibrillation thresholds before and after biventricular subendocardial ablation were 25 +/- 3 and 22 +/- 10 mA, respectively (p = NS). CONCLUSIONS: We conclude that 1) subendocardial ablation does not decrease ventricular vulnerability when the heart is in situ and is not on cardiopulmonary bypass and 2) anodal supernormal excitability can be demonstrated in ventricles without a subendocardial Purkinje fiber network. PMID- 8419012 TI - Variable effects of explosive or gradual increase of intracranial pressure on myocardial structure and function. AB - BACKGROUND: Studies done in potential donors for heart transplantation and in experimental animals have suggested that brain death can have major histopathological and functional effects on the myocardium. METHODS AND RESULTS: We developed experimental models of brain death using dogs to study the hemodynamic and catecholamine changes, the extent of myocardial structural damage, and the recovery potential of donor hearts obtained from brain-dead donors. Brain death was caused by increasing the intracranial pressure (ICP) suddenly or gradually by injecting saline in an epidural Foley catheter. In a first series of experiments, dogs given a sudden rise in ICP (n = 5) showed a hyperdynamic response and a 1,000-fold increase in the level of epinephrine after brain death. Histology revealed 93 +/- 2% of the myocardium to be severely ischemic. Dogs given a gradual rise in ICP (n = 6) showed a lesser hyperdynamic response, almost 200-fold increase in the level of epinephrine after brain death, and mild ischemic damage to the myocardium (23 +/- 1%). In a second series, hearts obtained from brain-dead and non-brain-dead donors were transplanted in recipients, and the weaning and recovery potential were studied. All four recipients with hearts from non-brain-dead donors were weaned with good functional recovery. Also, all four recipients with hearts from brain-dead dogs given a gradual rise in ICP were weaned with only moderate functional recovery. However, only two of four recipients with hearts from donors given a sudden rise in ICP were weaned and showed poor functional recovery. CONCLUSIONS: Our results indicate that a sudden rise in ICP can cause irreversible myocardial damage. PMID- 8419013 TI - Distribution of left ventricular sympathetic afferents demonstrated by reflex responses to transmural myocardial ischemia and to intracoronary and epicardial bradykinin. AB - BACKGROUND: Stimulation of left ventricular (LV) receptors with sympathetic afferents generally results in reflex sympathoexcitatory responses. Stimulation of LV receptors with vagal afferents results in reflex sympathoinhibitory responses. Vagal afferents are known to be preferentially distributed to the inferoposterior (IP) wall of the LV. We tested the hypothesis that there is also a preferential distribution of LV sympathetic afferents. METHODS AND RESULTS: We measured reflex responses to stimulation of sympathetic afferents located in the anterior and IP LV: We used myocardial ischemia and chemical stimuli to increase the activity of the sensory endings in 15 chloralose-anesthetized, mechanically ventilated dogs with sinoaortic denervation and vagotomy. Reflex responses were assessed by direct recordings of efferent renal sympathetic nerve activity (RSNA). In nine dogs, maximal RSNA changes elicited by transmural anterior myocardial ischemia (22.6 +/- 3.9% increase from baseline nerve traffic) were not significantly different from maximal RSNA changes observed during transmural IP ischemia (27.1 +/- 4.4%). Similar changes in mean arterial and left atrial pressures were noted during transmural anterior and IP ischemia. In eight dogs, maximal changes of RSNA elicited by epicardial or intracoronary bradykinin to the anterior LV were not significantly different from those observed during bradykinin to the IP LV (anterior epicardial bradykinin, 76.7 +/- 11.7%; IP epicardial bradykinin, 72.2 +/- 10.0%; anterior intracoronary bradykinin, 84.6 +/ 21.0%; IP intracoronary bradykinin, 88.8 +/- 17.3%). CONCLUSIONS: We conclude that cardiac receptors with sympathetic afferents are distributed equally to the IP and anterior regions of the LV. PMID- 8419014 TI - Altered left ventricular chamber stiffness and isovolumic relaxation in dogs with chronic pulmonary hypertension caused by emphysema. AB - BACKGROUND: In chronic obstructive lung disease, a right to left ventricular septal shift that occurs as a consequence of right ventricular pressure overload is the usual mechanism given to explain a decrease in left ventricular (LV) diastolic performance. The purpose of the present study was to examine the extent to which this mechanism could account for a decrease in LV diastolic function in a canine model in which pulmonary artery pressure was elevated to a level found in human disease. METHODS AND RESULTS: Severe emphysema was produced in dogs by repeated instillations of the enzyme papain into the lung. To assess LV diastolic function, we used sonomicrometry, in which three pairs of subendocardial crystal transducers were implanted along the three orthogonal axes of the LV. LV end diastolic dimensions and pressure-strain relations along the three axes, as well as the time constant of LV isovolumic relaxation (T), were measured before (baseline) and after 1 year of emphysema (post-1-year study). The results showed that after 1 year of pulmonary hypertension, LV pressure-strain relations were decreased along the septal-lateral and anterior-posterior axes, but a right to left ventricular septal shift was not detected. The relation of average midwall circumferential stress to midwall circumferential strain was used to describe the intrinsic compliance of the LV. The results showed that myocardial stiffness increased in emphysema but that chamber volume was not reduced. At the post-1 year study, T was abnormally increased in the emphysema group in response to augmented preload and afterload compared with preemphysema measurements. CONCLUSIONS: We conclude that mechanisms other than ventricular interdependence may be operative in leading to altered LV diastolic filling in chronic emphysema. PMID- 8419015 TI - Inhibition of accelerated coronary atherosclerosis with dehydroepiandrosterone in the heterotopic rabbit model of cardiac transplantation. AB - BACKGROUND: Accelerated coronary atherosclerosis has become a critical problem in cardiac transplantation. Although the pathogenesis of this disease is unknown, hypercholesterolemia has been shown to be a major risk factor. METHODS AND RESULTS: To study this problem, a hypercholesterolemic rabbit model of heterotopic cardiac transplantation was developed to study accelerated graft atherosclerosis. Based on suggestions in the literature, it was hypothesized that dehydroepiandrosterone (DHEA) may retard the progression of the disease. Using semiquantitative light microscopy, a predilection for the development of small vessel occlusive disease in the transplanted hearts was found. Chronic DHEA administration produced a 45% reduction in the number of significantly stenosed vessels in the transplanted hearts (p < 0.05) compared with controls and a 62% reduction in the nontransplanted hearts (p < 0.05), yielding an overall 50% reduction in the number of significantly stenosed vessels in both the transplanted and nontransplanted hearts. This reduction in luminal stenosis was observed in the absence of any significant alterations in lipid profiles. CONCLUSIONS: It is concluded that chronic DHEA administration in a hypercholesterolemic rabbit model of heterotopic cardiac transplantation significantly retards the progression of accelerated atherosclerosis in both the transplanted heart and in the native heart. PMID- 8419016 TI - Tricarboxylic acid cycle activity in postischemic rat hearts. AB - BACKGROUND: Although myocardial oxidative tricarboxylic acid (TCA) cycle activity and contractile function are closely linked in normal cardiac muscle, their relation during postischemic reperfusion, when contractility often is reduced, is not well defined. METHODS AND RESULTS: To test the hypothesis that oxidative TCA cycle flux is reduced in reperfused myocardium with persistent contractile dysfunction, TCA cycle flux was measured by analyzing the time course of sequential myocardial glutamate labeling during 13C-labeled substrate infusion with 13C nuclear magnetic resonance spectroscopy in beating isolated rat hearts at 37 degrees C. Total TCA cycle flux, indexed by both empirical and mathematical modeling analyses of the 13C data, was not reduced but rather increased in hearts reperfused after 17-20 minutes of ischemia (left ventricular pressure, 73 +/- 5% of preischemic values) compared with flux in developed pressure-matched controls (e.g., total flux, 2.5 +/- 0.4 versus 1.6 +/- 0.1 mumol.min-1.g wet wt-1, respectively; p < 0.01). No TCA cycle activity was detectable by 13C nuclear magnetic resonance in hearts reperfused after 40-45 minutes of ischemia, which lacked contractile recovery and had ultrastructural evidence of irreversible injury. CONCLUSIONS: These results suggest that TCA cycle activity is not persistently decreased in dysfunctional reperfused myocardium after a brief ischemic episode and therefore cannot account for the reduced contractile function at that time. PMID- 8419017 TI - Vascular injury induces angiotensinogen gene expression in the media and neointima. AB - BACKGROUND: Angiotensin II promotes growth of vascular smooth muscle cells in vitro via the autocrine production of growth factors such as platelet-derived growth factor, basic fibroblast growth factor, and transforming growth factor beta. Furthermore, experimental studies have demonstrated that angiotensin infusion can enhance smooth muscle proliferation after balloon injury in vivo. Consistent with this, angiotensin converting enzyme inhibitors have been shown to prevent myointimal proliferation. The origin of vascular angiotensin that participate in this process is of interest. We have demonstrated the presence of angiotensinogen messenger RNA (mRNA) in the adventitial and medial layers of the rat aorta and have speculated that local angiotensinogen production may play an important role during myointimal proliferation. To provide further evidence toward this hypothesis, we compared the localization and expression of angiotensinogen mRNA in control and balloon injured vessels using in situ hybridization. METHODS AND RESULTS: Abdominal aorta of Sprague-Dawley rats were studied before or after injury with a balloon catheter. Neointimal hyperplasia developed as documented morphologically by a progressive increase in the ratio of neointimal to medial thickness from 0.17 at 1 week to 1.17 at 6 weeks after injury. Angiotensinogen mRNA was detected clearly in the adventitia and media of control and injured aorta. However, at 1 week after injury, the medial-to adventitial angiotensinogen mRNA ratio was higher in the injured aorta, suggesting increased gene expression in the media compared with control. Of potential importance, angiotensinogen mRNA was also detected in the neointima of the injured aorta, and this was also highest at 1 week after injury. CONCLUSIONS: These data are consistent with the hypothesis that balloon injury leads to activation of the vascular renin-angiotensin system, which may participate in the myointimal proliferation. PMID- 8419018 TI - Prosthetic heart valve thrombosis. 'What can be done with regard to treatment?'. PMID- 8419019 TI - Smoking after acute myocardial infarction. A good thing? PMID- 8419020 TI - The St. Jude valve. Thrombolysis as the first line of therapy for cardiac valve thrombosis. AB - BACKGROUND: Thrombolytic therapy is a promising alternative to valve replacement in the management of prosthetic valve thrombosis. We sought to determine the short- and long-term results of treating thrombosed St. Jude heart valves with thrombolytic therapy as the primary treatment modality. METHODS AND RESULTS: Between March 1978 and December 1991, 988 patients underwent implantation of St. Jude prosthetic valves at our medical center, and all patients with thrombosed valves were identified prospectively. During this period, 17 patients (13 women; mean age, 66.8 +/- 19.0 years) developed prosthetic valve thrombosis (11 aortic, six mitral). In six patients, Coumadin was stopped in preparation for elective surgery. The clinical presentation was congestive heart failure in 13, syncope and fatigue in two, and a cerebrovascular accident in one; one patient was asymptomatic. The average duration of symptoms was 11.7 +/- 12.0 days (range, 1 45 days). Anticoagulation was subtherapeutic in all but one patient at the time of presentation. Cinefluoroscopy was the primary method used for diagnosis and was also used to follow the response to therapy. Twelve patients were treated medically (10 with thrombolytic therapy and two with heparin), three were treated surgically, and two were diagnosed at autopsy. Of the 12 medically treated patients, 10 had marked improvement in leaflet movement and symptoms within 12 hours. Thus, 10 of 12 patients (83%) had a satisfactory response to medical therapy alone. No medically treated patient died or had a major complication resulting in permanent damage. However, four of the 12 medically treated patients had minor complications, including a transient episode of facial weakness in one patient, hematomas in two, and epistaxis in one. Late rethrombosis recurred in two patients in the medically treated group and was successfully retreated with thrombolytic therapy. At 3 months, all patients were alive and well. CONCLUSIONS: Thrombolytic therapy can be used as the first line of therapy for thrombosed St. Jude valves with a low risk of permanent side effects and excellent chances of success. In most cases, surgery can be reserved for patients who do not respond to thrombolytic therapy. PMID- 8419021 TI - Is OT interval prolongation a strong or a weak predictor for cardiac death? PMID- 8419022 TI - Joint effects of serum triglycerides and LDL and HDL cholesterol. PMID- 8419023 TI - Early effects of tissue-type plasminogen activator added to conventional therapy on the culprit coronary lesion in patients presenting with ischemic cardiac pain at rest. Results of the Thrombolysis in Myocardial Ischemia (TIMI IIIA) Trial. AB - BACKGROUND: The early effects of tissue-type plasminogen activator (t-PA) on the "culprit" coronary lesion in patients presenting with unstable angina or non-Q wave myocardial infarction were determined by quantitative arteriography. METHODS AND RESULTS: Of 391 such patients, 306 satisfied clinical and arteriographic requirements for eligibility and received a 90-minute front-loaded infusion of t PA (0.8 mg/kg i.v.; maximum, 80 mg) or placebo plus conventional antianginal therapy. All patients received full heparinization and a follow-up arteriogram 18 48 hours after treatment. A non-Q wave myocardial infarction (MI) was diagnosed in 97 patients (32%) after entry. In the entire patient population, among t-PA- and placebo-treated patients, respectively, 25% versus 19% (p = 0.25) of all culprit lesions achieved the primary study end point, measurable improvement (by > or = 10% reduction of stenosis or two Thrombolysis in Myocardial Infarction [TIMI] flow grades) at follow-up. Substantial improvement (by > or = 20% reduction of stenosis or two TIMI grades) was seen with t-PA in 15% of all culprit lesions versus 5% with placebo (p < 0.003). Arteriographically apparent thrombus was present at baseline in the culprit lesion of 107 patients (35%). Substantial improvement was more frequent with t-PA among lesions containing apparent thrombus (in 36% with t-PA versus 15% with placebo; p < 0.01), as it was among patients evolving a non-Q wave MI (33% versus 8%; p < 0.005). By multivariate analysis, the significant, independent predictors of substantial improvement include apparent thrombus (p = 0.0001), non-Q wave MI (p = 0.003), and t-PA use (p = 0.01). Both non-Q wave MI status and thrombus had been specified a priori as important variables. CONCLUSIONS: Arteriographically apparent intraluminal thrombus and improvement of the culprit lesion with either of these regimens were only moderately frequent in patients with unstable angina or non-Q wave MI. Substantial improvement of culprit lesions was more frequent with t-PA than with placebo overall and in two prospectively defined subgroups. The clinical relevance of these observations is being tested in the larger, ongoing clinical TIMI IIIB study. PMID- 8419024 TI - Significance of smoking in patients receiving thrombolytic therapy for acute myocardial infarction. Experience gleaned from the International Tissue Plasminogen Activator/Streptokinase Mortality Trial. AB - BACKGROUND: Despite the fact that smoking is a well-established risk factor for the development of coronary artery disease, some investigators have noted that hospital mortality after acute myocardial infarction is lower in patients who smoke than in nonsmoking patients. To evaluate the association of smoking with mortality during hospitalization after thrombolytic therapy and 6 months afterward, we analyzed the results of the International Tissue Plasminogen Activator/Streptokinase Mortality Trial. METHODS AND RESULTS: Patients were divided into three groups: nonsmokers (those who never smoked), ex-smokers, and active smokers. Multivariate and univariate comparisons were made with respect to baseline characteristics and clinical outcome. There were 2,366 nonsmokers, 2,244 ex-smokers, and 3,649 active smokers. The baseline characteristics of nonsmoking patients differed significantly from the ex-smokers and active smokers. The nonsmoking group included more women than the ex-smokers or active smokers (45% versus 10.6% and 17.6%, respectively), was older (67 +/- 10 years versus 64 +/- 10 years and 58 +/- 11 years), had a higher rate of diabetes mellitus (16.3% versus 11.1% and 7.5%), and had a worse Killip class at admission. Nonsmoking patients and ex-smokers experienced more in-hospital reinfarction than active smokers (4.7% and 5% versus 2.7%, p < 0.0001, respectively). Nonsmokers experienced more in-hospital shock than the ex-smokers or active smokers (9.2% versus 6.4% and 5.8%, p < 0.0001), stroke (1.9% versus 1.8% and 0.8%, p < 0.0001), and bleeding (7.2% versus 6.5% and 4.4%, p < 0.0001). They also experienced a higher in-hospital and 6-month mortality (12.8% and 17.6%) than ex smokers (8.2% and 12.1%) or active smokers (5.4% and 7.8%) (p < 0.0001). A multivariate analysis accounting for all baseline characteristics demonstrated a significant association between nonsmoking and increased hospital mortality, with an odds ratio of 1.42 (confidence limits, 1.15-1.72). Among active smokers, there was a nonsignificant trend for mortality rates to decrease with increasing numbers of cigarettes smoked per day. CONCLUSIONS: This retrospective analysis indicates that smokers receiving thrombolytic therapy after acute myocardial infarction have significantly better hospital and 6-month outcome than nonsmokers or ex-smokers. However, smokers sustained their infarction at a significantly earlier age than nonsmokers, and strenuous efforts should continue to be made to decrease the incidence of new and continued smoking. PMID- 8419025 TI - Comparison of hemodynamic determinants for myocardial oxygen consumption under different contractile states in human ventricle. AB - BACKGROUND: Recently, several indexes such as tension-time index (TTI), tension time or force-time integral (FTI), rate-pressure product (RPP), pressure-work index (PWI), and systolic pressure-volume area (PVA) have been developed as predictors of myocardial oxygen consumption in experimental and clinical studies. However, it is still unclear whether these indexes are reliable predictors of myocardial oxygen consumption under various contractile states in human hearts. METHODS AND RESULTS: We assessed the relation between TTI, FTI, RPP, PWI, and PVA and myocardial oxygen consumption per beat (VO2) in 13 patients with heart disease during volume loading. Left ventricular (LV) volume and pressure were measured simultaneously by the conductance catheter with the tipped micromanometer technique. VO2 was calculated from arterial coronary sinus oxygen content difference, and coronary sinus blood flow was measured by the thermodilution method. After z transformation of the correlation coefficients, mean z value for the VO2-PVA relation (1.83 +/- 0.60) was greater than those for the VO2-TTI relation (1.22 +/- 0.66; p < 0.005), VO2-FTI relation (1.18 +/- 0.61; p < 0.05), VO2-RPP relation (0.95 +/- 0.65; p < 0.05), and VO2-PWI relation (1.24 +/- 0.58; p < 0.05). During dobutamine infusion (5 micrograms.kg-1.min-1) in five of the 13 patients, VO2 also correlated best with PVA (z = 1.70 +/- 0.89) compared with TTI (z = 1.43 +/- 0.86), FTI (z = 1.48 +/- 0.95), RPP (z = 1.00 +/- 0.53), and PWI (z = 0.88 +/- 0.80). The contractile efficiency (38 +/- 14% to 38 +/- 20%), the reciprocal of the slope of the VO2-PVA relation, remained unchanged, whereas the VO2,PVA 0.8 (VO2 at PVA = 0.8 J per beat/100 g LV) increased from 1.48 +/- 1.16 to 2.06 +/- 1.13 J per beat/100 g LV (p < 0.05). These results show the parallel upward shift of the VO2-PVA relation during dobutamine infusion. Because increases in the VO2-intercept represent the VO2 for the increased excitation-contraction (E-C) coupling associated with the augmented contractile state, the parallelism of the VO2-PVA relation could discriminate between VO2 for mechanical work (PVA-dependent VO2) and VO2 for E-C coupling (PVA independent VO2). CONCLUSIONS: The results of the present study indicate that PVA is a reliable and valuable predictor of myocardial oxygen consumption under different contractile states in human hearts. The VO2-PVA relation could provide useful information about mechanoenergetics in diseased human hearts. PMID- 8419026 TI - Cigarette smoking is a major risk factor for coronary spasm. AB - BACKGROUND: Although there have been many studies on the risk factors for coronary artery disease, the etiology of coronary artery spasm has not yet been determined. METHODS AND RESULTS: After diagnosis by coronary arteriography, various risk factors were compared between two groups of subjects using logistic regression analysis. The vasospasm group included 175 patients with angiographically determined coronary artery spasm but no coronary artery narrowing exceeding 25% of the luminal diameter. The control group comprised 176 subjects with completely normal coronary arteries and a negative response to ergonovine maleate. The adjusted odds ratio and 95% confidence interval for smoking as a risk factor for vasospasm was 2.41 and 1.53-3.82, respectively (p < 0.05). The adjusted odds ratios for total cholesterol, low density lipoprotein, high density lipoprotein, triglycerides, diabetes mellitus, and body mass index, calculated by multivariate logistic regression analysis, were not statistically nonsignificant. CONCLUSIONS: Smoking appears to be a major risk factor for vasospastic angina without significant coronary narrowing. The other risk factors for coronary artery disease may not contribute to coronary vasospasm. PMID- 8419027 TI - Hemoglobin inhibits endothelium-dependent relaxation to acetylcholine in human coronary arteries in vivo. AB - BACKGROUND: The endothelium can regulate vascular tone by releasing both endothelium-derived relaxing factor (EDRF or nitric oxide) and contracting factors. To date, there has only been circumstantial evidence to indicate EDRF activity in vivo in human coronary arteries. Using human hemoglobin as a specific inhibitor, the hypothesis that acetylcholine-induced coronary vasodilation is due to EDRF release was tested. METHODS AND RESULTS: We studied the response of normal coronary arteries to acetylcholine (an endothelium-dependent vasodilator) and isosorbide dinitrate (an endothelium-independent vasodilator) in seven patients. The specificity of any vasodilator response was assessed by the infusion of reduced free human hemoglobin. Hemoglobin 10(-5) M infusion alone had no effect on coronary artery diameter. Drugs were infused into the coronary artery, and the diameter changes were assessed by quantitative angiography. Acetylcholine 10(-7) M increased left anterior descending coronary artery diameter from control: 2.30 +/- 0.12 mm to 2.79 +/- 0.20 mm (mean +/- SEM, n = 7, p < 0.01). Hemoglobin both in a concentration of 10(-6) M and 10(-5) M reversed this vasodilator effect, causing constriction to 2.11 +/- 0.18 mm (p < 0.001 compared with acetylcholine 10(-7) M) and 2.29 +/- 0.14 mm (p < 0.05 compared with acetylcholine 10(-7) M). Isosorbide dinitrate in the presence of hemoglobin caused dilatation of the coronary artery in all cases to 3.04 +/- 0.24 mm (p < 0.001 compared with acetylcholine 10(-7) M and hemoglobin 10(-6) M). CONCLUSIONS: Using a specific inhibitor of nitric oxide, reduced free hemoglobin, we have demonstrated that basal EDRF release does not appear to play an important role in the maintenance of human epicardial coronary artery diameter in vivo but is responsible for the acetylcholine-induced dilatation. PMID- 8419028 TI - Hypertension and left ventricular hypertrophy are associated with impaired endothelium-mediated relaxation in human coronary resistance vessels. AB - BACKGROUND: Patients with hypertension and myocardial hypertrophy may have signs and symptoms of myocardial ischemia in the absence of obstructive coronary disease. Prior investigations have demonstrated impaired coronary flow reserve and have led to speculation that microvascular dysfunction might contribute to ischemia in these patients. Experimental studies have shown that the endothelium, an important regulator of microvascular tone, can be damaged by hypertension and is dysfunctional in cardiomyopathy. We hypothesized that endothelium-dependent vasodilation is impaired in the coronary microvasculature of patients with hypertension and ventricular hypertrophy. METHODS AND RESULTS: We studied coronary microvascular responses in 10 patients with left ventricular hypertrophy secondary to essential hypertension (HTN) (mean arterial pressure at catheterization, 151/94 mm Hg; mean posterior wall thickness, 1.4 +/- 0.1 cm) and nine normal control subjects with no history of hypertension (mean arterial pressure at catheterization, 128/75 mm Hg; mean posterior wall thickness, 1.0 +/- 0.02 cm) using the intracoronary Doppler catheter and quantitative angiography to assess changes in coronary blood flow (CBF). All patients had normal left ventricular systolic function. To assess microvascular endothelial function, we infused the endothelium-dependent vasodilator acetylcholine (10(-8)-10(-6) M) and the endothelium-independent vasodilator adenosine (10(-6)-10(-4) M) into the left anterior descending coronary artery. In response to acetylcholine, CBF increased only 32 +/- 25% in HTN patients, whereas CBF increased 192 +/- 39% in normal control subjects (p = 0.003). CBF increased 465 +/- 93% in HTN patients and 439 +/- 41% in normal control subjects in response to adenosine (p = NS). The proportion of coronary flow reserve attributable to endothelium-dependent dilation (obtained from peak acetylcholine/peak adenosine flow response) was 48 +/- 9% in normal control subjects but only 7 +/- 8% in HTN patients (p = 0.008). CONCLUSIONS: Endothelium-dependent vasodilation is markedly impaired in the coronary microvessels of patients with hypertension and ventricular hypertrophy. Loss of this vasodilator mechanism may contribute to disordered coronary flow regulation and the ischemic manifestations of hypertensive heart disease. PMID- 8419029 TI - Fat-suppressed breath-hold magnetic resonance coronary angiography. AB - BACKGROUND: The ability to image the coronary arteries noninvasively would represent an advance in patient care. We have developed a magnetic resonance (MR) angiographic technique that allows the acquisition of complete images of coronary flow within a single breath-hold. By this method, the feasibility of noninvasive MR coronary angiography was evaluated in 25 subjects, including 19 healthy adult volunteers and six patients after diagnostic coronary angiography. METHODS AND RESULTS: Noninvasive MR coronary angiography was performed with a fat-suppressed ECG-gated gradient-echo sequence with k-space segmentation. Overlapping transverse sections were initially used to image coronary flow, with oblique images obtained after identification of proximal anatomy. The left main coronary artery was seen in 24 subjects (96%), with a mean diameter of 4.8 mm (range, 3.4 6.2 mm) and average length of 10 mm (range, 8-14 mm). The left anterior descending coronary artery was seen in 100% of subjects, with a mean proximal diameter of 3.6 mm (range, 2.6-4.3 mm) and for an average length of 44 mm (range, 28-93 mm). The left circumflex coronary artery was seen in 76% of subjects, with a mean proximal diameter of 3.5 mm (range, 2.6-4.3 mm) and for an average length of 25 mm (range, 9-42 mm). The right coronary artery was also identified in 100% of subjects, with a mean proximal diameter of 3.7 mm (range, 2.7-5.1 mm) and for an average length of 58 mm (range, 24-122 mm). Quantitative angiography of normal proximal segments demonstrated a good correlation with MR-determined lumen diameters (r = 0.86, p < 0.002). Occluded vessels in patients with coronary artery disease displayed an absence of flow signal distal to the occlusion, whereas vessels with significant angiographic stenoses demonstrated signal loss corresponding to the area of the stenosis, with visualization of the more distal vessel. CONCLUSIONS: Breath-hold MR coronary angiography provides visualization of the major epicardial vessels. In the future, MR coronary angiography may provide a noninvasive means for the evaluation of patients with known or suspected coronary artery disease. PMID- 8419030 TI - Analytical performance evaluation of EMIT II monoclonal amphetamine/methamphetamine assay: more specificity than EMIT d.a.u. monoclonal amphetamine/methamphetamine assay. AB - We evaluated a new EMIT II monoclonal amphetamine/methamphetamine assay for screening human urine by comparing it with the EMIT d.a.u. monoclonal amphetamine/methamphetamine assay and a fluorescence polarization assay. The EMIT II assay has a cutoff of 1 mg/L d-methamphetamine. The EMIT II and EMIT d.a.u. assays were run on a BM/Hitachi 704 analyzer; for the fluorescence polarization assay we used a TDx analyzer. All EMIT II positive samples were also analyzed by the fluorescence polarization assay. We used gas chromatography/mass spectrometry (GC/MS) for confirmation of the presence of amphetamine or methamphetamine. Within-run CVs for the Level 1 (1 mg/L) and Level 2 (3 mg/L) calibrators for the EMIT II assay were 0.47% and 0.53%, respectively. Corresponding between-run CVs were 1.48% and 1.60%, respectively. Of the 1007 samples screened for amphetamines, 50 were positive by the EMIT d.a.u. assay; 21 samples (not a subset of the 50 samples) were positive by the EMIT II assay. However, 19 samples that tested positive by EMIT II also tested positive by the EMIT d.a.u. assay. Subsequent testing of the EMIT II positive samples by the fluorescence polarization assay detected in six positive samples. By means of chiral derivatization wer identified two specimens containing primarily l-isomers of amphetamine and methamphetamine. Sympathomimetic amines were identified in several of the samples not containing amphetamine or methamphetamine. PMID- 8419031 TI - Phenol and p-cresol accumulated in uremic serum measured by HPLC with fluorescence detection. AB - We developed a simple and sensitive high-performance liquid chromatographic (HPLC) method that uses fluorescence as a detector for quantifying serum phenol and p-cresol in uremic patients on hemodialysis. Identification of phenol and p cresol was confirmed by liquid chromatography/mass spectrometry. Because the HPLC method requires only simple extraction by ethyl acetate and does not require further steps such as derivatization, it is simple and rapid compared with gas chromatography or gas chromatography/mass spectrometry. Concentrations of phenol and p-cresol in uremic serum were significantly (p < 0.01) higher than those in normal serum. Reduction rates of phenol and p-cresol by hemodialysis were lower than those of urea and creatinine, suggesting a protein-binding property of phenol and p-cresol. This method will be useful for monitoring serum phenols in dialyzed patients as an index of hemodialysis adequacy. PMID- 8419032 TI - Evaluation of silyl-blocked p-nitrophenylmaltoheptaoside as a substrate for alpha amylase reagents. AB - We describe a reagent for measuring alpha-amylase (EC 3.2.1.1) activity in serum with use of a thexyldimethylsilyl ether of p-nitrophenyl-alpha-D-maltoheptaoside (SB7) as substrate. This substrate differs from Genzyme's benzylidene-blocked p nitrophenylmaltoheptaoside substrate (B-PNPG7). The reagent, optimized for the characteristics of the silyl-blocked substrate, contains 4-(2-hydroxyethyl)-1 piperazineethane sulfonate buffer at pH 7.3, alpha-glucosidase (maltase; EC 3.2.1.20), and glucoamylase (EC 3.2.1.3). Comparison with Ciba Corning Diagnostics Corp.'s, amylase reagent with B-PNPG7 as substrate (x) yielded a regression equation of y = 1.20x-2.7 (r = 0.9997). The linear range exceeded amylase concentrations > 2500 U/L and total precision (CV) was 2.3% at an amylase concentration of 112 U/L with the Ciba Corning 550 Express analyzer. Reconstituted reagent is stable for 30 days at 5 degrees C and 7 days at ambient (18-25 degrees C) temperatures. PMID- 8419033 TI - Cyclosporin G and metabolite binding to cyclophilin and a 50-kDa binding protein related to in vitro immunosuppression. AB - Seven purified metabolites of cyclosporin G (CsG) were studied for binding to cyclophilin and a 50-kDa binding protein (50-kDa BP). The ratios of the metabolite dissociation constants with respect to CsG were compared with in vitro immunosuppression by using the primary mixed lymphocyte suppression assay. The immunosuppressive potency ratio of the parent compounds, both cyclosporin A (CsA) and CsG, compared favorably with the drug dissociation constants for cyclophilin and the 50-kDa BP. Three of the seven metabolites had comparable binding and potency ratios for the 50-kDa BP. In contrast, none of the seven metabolites appreciably bound to cyclophilin in the concentration range tested. PMID- 8419034 TI - Discordant results for determinations of triglycerides in pig sera. AB - We recently determined triglyceride concentrations in pig sera by three fully enzymatic methods (Kodak Ektachem 700, Hitachi 707, and Abbott EPx) and obtained significantly lower values than those obtained with chemical or enzymatic methods based on chemical hydrolysis. All methods used involve microbial lipases for liberating glycerol from glycerides and glycerol phosphate dehydrogenases or oxidases for subsequent oxidation. The methods were validated against reference methods by using fresh human sera and survey materials. The discordant results were not from matrix sample-method interaction but from incomplete hydrolysis of pig serum triglycerides by the lipolytic enzymes. When serum triglycerides from 10 pigs showing the highest biases were hydrolyzed by microbial lipases and the reaction mixture was subjected to thin-layer and gas-liquid chromatography, the predominant end products were palmitoyl monoglyceride and a mixture of free fatty acids with the following composition (fatty acid as percent of total +/- SD): 16:0, 7.8 +/- 2; 18:0, 5.4 +/- 2.2; 18:1, 53 +/- 12; 18:2, 31 +/- 4.6; and 18:3, 2.5 +/- 1. Assuming that the lipases exhibit the usual specificity toward the 1 and 3 positions of the triglyceride, the data suggest that, in pig, triglycerides 18:1 and 18:2 occupy the 1 and 3 positions and 16:0 (palmitic acid) predominantly occupies the 2 position. Triglycerides of this structure may not be well hydrolyzed by the typical lipolytic enzymes in clinical assays. PMID- 8419036 TI - Medical-care systems for long-duration space missions. AB - As in the opening of frontiers on Earth, human physiological maladaptation, illness, and injury--rather than defective transportation systems--are likely to be the pace-limiting variables in efforts to expand the presence of humans into the solar system. Because of the inability of individuals to return to Earth rapidly and conveniently, the capability of delivering medical care on site will be key to the success of a manned space station, lunar base, and Mars mission. Spaceflight medical care equipment must meet stringent constraints of size, weight, and power requirements, and then must function accurately in remote, self contained, microgravity settings after extended intervals of storage, with neither expert operators nor repair technicians on site. Satisfying these unusually rigorous requirements will require sustained direct involvement of clinically up-to-date health-care providers, medical scientists, and biomedical engineers, as well as astronauts and aerospace engineers and managers. Solutions will require validation in clinical settings with real patients, as well as in simulated operational settings. PMID- 8419035 TI - Phenylalanine determined in plasma with use of phenylalanine dehydrogenase and a centrifugal analyzer. AB - Quantitative determination of plasma phenylalanine (Phe) is essential for the diagnosis of phenylketonuria (PKU) and the control of dietary therapy of PKU patients. We have adapted a spectrophotometric method (Clin Chim Acta 1991;201:95 8) based on phenylalanine dehydrogenase (EC 1.4.1.-) for rapid and accurate determinations of Phe with the Cobas Fara II centrifugal analyzer. The method is based on the NAD(H)-dependent reductive deamination of Phe with large amounts of phenylalanine dehydrogenase, which catalyzes the formation of phenylpyruvate at pH 10.8. Combining the specificity of the phenylalanine dehydrogenase with the precision, accuracy, and considerable time-saving of an automated system is useful for monitoring PKU patients in a children's hospital. PMID- 8419037 TI - Biochemical markers for detecting bone metastases in patients with breast cancer. AB - A study was carried out to assess the best use of biochemical bone markers to exclude metastases in patients with breast cancer. Urinary galactosyl hydroxylysine and serum alkaline phosphatase were used to monitor bone resorption and deposition, respectively. Hydroxyproline was also measured. In a selected population of patients, possibly affected by metastases on the basis of scintigraphic examination, which is highly sensitive but poorly specific, we assessed the efficiency of the markers by a double statistical analysis. In this group, the only marker able to predict metastases was galactosyl-hydroxylysine. PMID- 8419038 TI - Plasma and urinary oxalate and glycolate in healthy subjects. AB - High-performance ion chromatography (HPIC) has been widely used for oxalate analysis and, more recently, for glycolate analysis. We describe a procedure for sample preparation in which the plasma ultrafiltrate is acidified during harvesting with a cation-exchange resin, and the chloride is removed before the ion chromatography, which is performed with a newly developed AS10 column. The same ultrafiltrate sample is analyzed for glycolate. For plasma oxalate, the mean recovery of sample in eluted fractions was 95-96%, and intraassay CV was 6.2 8.1%. The reference interval (mean +/- 2 SD) for men was 0.8-3.2 mumol/L and for women, 1.0-2.6 mumol/L. For urinary oxalate, the reference interval for men was 175-560 mumol/day and for women, 107-432 mumol/day. For plasma glycolate, the mean analytical recovery was 96-98%, and the intra-assay CV was 2.4-6.2%. The reference interval for men was 1.9-7.5 mumol/L and for women, 1.4-7.4 mumol/L. For urinary glycolate, the reference interval for men was 0-1400 mumol/day and for women, 91-1001 mumol/day. PMID- 8419039 TI - Interference of carbamylated and acetylated hemoglobins in assays of glycohemoglobin by HPLC, electrophoresis, affinity chromatography, and enzyme immunoassay. AB - In vitro-synthesized carbamylated and acetylated hemoglobins interfered in assays of glycohemoglobin by HPLC and electrophoresis but had no effects on results obtained by affinity chromatography and enzyme immunoassay. Correlations between long-term serum urea concentrations and glycohemoglobin percentages revealed that, in vivo, carbamylated hemoglobin equivalent to 0.063% of total hemoglobin is formed for every 1 mmol/L of serum urea. The use of acetylsalicylate, either chronically in small doses (200-300 mg/day) or for 1 week at 2000 mg/day, did not cause significant interference from acetylhemoglobin, formed in vivo. We conclude that interference from carbamylated hemoglobin explains only a small part of existing discrepancies between results of glycohemoglobin assays in current use. The interfering effect of acetylhemoglobin formed in vivo with acetyl-CoA as substrate is as yet unknown. PMID- 8419040 TI - Robotic chromatography: development and evaluation of automated instrumentation for assay of glycohemoglobin. AB - The measurement of glycohemoglobin (GHb) by boronate affinity chromatography is useful in monitoring long-term glucose control in diabetic subjects. The inherent disadvantage of this method is the hands-on time required because the hemoglobin fractions are separated on individual disposable columns. To overcome this disadvantage, we have programmed a Hamilton Microlab 2200 automated pipetting cartesian robot to complete the procedure, from the aspiration of blood from the sample-collection tube to the transfer of the separated hemoglobin fractions to a microtiter plate for absorbance measurement. This automated robotic system can analyze 96 specimens, including patients' samples and control material, in approximately 3 h. The precision (CV) of the method ranged from 1.6% to 3.5% within-run and from 2.7% to 3.5% day-to-day. The results correlated with those obtained with the Accuflex semiautomated robot, which used the identical disposable column, and those obtained with a Primus high-performance liquid chromatograph, which used a regenerated microparticle column. Automation of the GHb procedure allowed improved throughput, reduced labor cost, improved precision, and offered greater laboratory safety. PMID- 8419041 TI - C-reactive protein and its cytokine mediators in intensive-care patients. AB - C-reactive protein (CRP) is an acute-phase protein produced by the liver during bacterial infections and inflammation. The cytokines interleukin (IL)-1 beta, IL 6, and tumor necrosis factor (TNF) are widely reported to induce synthesis of CRP by hepatocytes both in vitro and in vivo. We investigated the relation between CRP and its cytokine mediators in 64 critically ill patients during their treatment in the intensive-care unit. Plasma CRP and IL-6 concentrations were significantly lower in patients without any evidence of infection than in those with clinical infection; plasma IL-1 beta concentrations showed no significant difference between any of the groups, but plasma TNF concentrations were lower in patients with evidence of infection. Significant correlation was seen between plasma concentrations of CRP and IL-6 when the latter was measured by bioassay; however, IL-6 showed, at best, only a 50% predictive value for a change in CRP concentration. PMID- 8419042 TI - Automated determination of lysozyme activities in biological fluids. PMID- 8419043 TI - The birth of the atomic absorption spectrometer and its early applications in clinical chemistry. PMID- 8419044 TI - Effects of icing whole-blood samples in plastic go beyond PO2 alone. PMID- 8419045 TI - Pitfalls of screening for mucopolysaccharidoses by the dimethylmethylene blue test. PMID- 8419046 TI - Propylene glycol interference in gas-chromatographic assay of ethylene glycol. PMID- 8419047 TI - Misleading use of correlation and regression in method-comparison studies of cyclosporine. PMID- 8419048 TI - Monitoring cyclosporine by HPLC with cyclosporin C as internal standard. PMID- 8419049 TI - HPLC micro-method for determining delta-aminolevulinic acid in plasma. PMID- 8419050 TI - Skeletal muscle phosphate uptake during euglycemic-hyperinsulinemic clamp. PMID- 8419051 TI - A minor improvement of the Limulus test. PMID- 8419052 TI - Determination of tetramethylenedisulfotetramine in blood by gas chromatography. PMID- 8419053 TI - Serum thyrotropin concentration in apparently healthy adults, in relation to urinary iodide concentration. PMID- 8419054 TI - Erroneous laboratory results from hemolyzed, icteric, and lipemic specimens. PMID- 8419055 TI - The clinical chemistry and immunology of long-duration space missions. AB - Clinical laboratory diagnostic capabilities are needed to guide health and medical care of astronauts during long-duration space missions. Clinical laboratory diagnostics, as defined for medical care on Earth, offers a model for space capabilities. Interpretation of laboratory results for health and medical care of humans in space requires knowledge of specific physiological adaptations that occur, primarily because of the absence of gravity, and how these adaptations affect reference values. Limited data from American and Russian missions have indicated shifts of intra- and extracellular fluids and electrolytes, changes in hormone concentrations related to fluid shifts and stresses of the missions, reductions in bone and muscle mass, and a blunting of the cellular immune response. These changes could increase susceptibility to space-related illness or injury during a mission and after return to Earth. We review physiological adaptations and the risk of medical problems that occur during space missions. We describe the need for laboratory diagnostics as a part of health and medical care in space, and how this capability might be delivered. PMID- 8419056 TI - Preliminary evaluation of an experimental clinical chemistry analyzer developed for space medicine. AB - An experimental clinical chemistry analyzer system was designed and built to demonstrate the feasibility of clinical chemistry as part of a medical-care system at NASA's planned space station Freedom. We report the performance of the experimental analyzer, called a medical development unit (MDU), for selected analytes in a laboratory setting in preparation for a preliminary clinical trial at patients' bedsides in an intensive-care unit. Within-run CVs ranged from 0.7% for sodium to 7.1% for phosphorus; day-to-day CVs ranged from 1.0% for chloride to 23.4% for calcium. Correlation of patients' blood sample analyses compared well with those by Ektachem E700 and other high-volume central laboratory analyzers (r ranged from 0.933 for creatine kinase MB isoenzyme to 0.997 for potassium), except for hemoglobin (r = 0.901) and calcium (r = 0.823). Although several CVs obtained in this study exceeded theoretical desired precision limits based on biological variations, performance was adequate for clinical laboratory diagnosis. We examined the effect of potentially interfering concentrations of hemoglobin, bilirubin, and lipids: the only effect was negative interference with calcium analyses by high concentrations of bilirubin. We also examined the effects of preanalytical variables and the performance of experimental sample transfer cups designed to retain sample and reference liquid in microgravity. Continued development of the MDU system is recommended, especially automation of sample processing. PMID- 8419057 TI - Ionic binding, net charge, and Donnan effect of human serum albumin as a function of pH. AB - The ionic activities and total molalities of sodium, potassium, calcium, lithium, and chloride in a solution of human serum albumin were measured at different values of pH between 4 and 9. The same quantities were measured simultaneously in a protein-free electrolyte solution in membrane equilibrium with the albumin solution. Taking the residual liquid-junction potential and bias from unselectivity of the electrodes into account, we determined the own, bound, and net charges of albumin. Chloride was amply bound at low pH, and calcium at high pH. The varying charge of ions bound to albumin opposed the effect of acid or base on the net charge. All ions were distributed across the membrane according to the same electric potential difference, which equalled the Donnan potential. The high concordance between observation and theory favors the Donnan theory and furthermore implies that the electrodes are as accurate in a solution with albumin as in a protein-free solution. PMID- 8419058 TI - Reliability and feasibility of pregnancy home-use tests: laboratory validation and diagnostic evaluation by 638 volunteers. AB - All 27 home-use tests sold in France in 1989 for the self-diagnosis of pregnancy were evaluated. The kits were first tested by qualified clinical chemistry technologists. Eleven kits with 100% specificity and 100% sensitivity were retained for the diagnostic study. Each of 638 laywomen was given a kit and asked to perform the assay with a coded urine specimen containing either no human chorionic gonadotropin (hCG) or an hCG concentration adjusted to the claimed detection limit (1 DL) or twice the detection limit (2 DL). After testing, each participant filled out a detailed questionnaire. The results showed a diagnostic specificity of 86-100% for 10 kits but a diagnostic sensitivity of 85-100% for only 5 kits at 2 DL and for only 2 at 1 DL. Among the 478 positive urine samples distributed, 230 were falsely interpreted as negative. The main explanation for such a high percentage of false-negative results was difficulty in understanding the explanatory leaflets accompanying the kits and hence in reading the results, regardless of the socioeconomic situation of the participant. We conclude that pregnancy home-use tests should be subjected to rigorous analytical controls and evaluated by a panel of potential users before being released on the market. PMID- 8419059 TI - Increase of plasma proapolipoprotein A-I in patients with liver cirrhosis and its relationship to circulating high-density lipoproteins 2 and 3. AB - We determine the concentration of proapolipoprotein (proapo) A-I and its ratio with total apolipoprotein (apo) A-I (proapo A-I/total apo A-I) in plasma of patients with liver disease; we used a noncompetitive sandwich method, an enzyme linked immunosorbent assay. The mean (SD) proapo A-I concentrations in patients with decompensated or compensated liver cirrhosis were higher than in normal subjects: 88 (25), 105 (36), and 69 (25) mg/L, respectively. The mean (SD) ratio (expressed as %) for each of these types of liver cirrhosis was also higher than in normal subjects: 10.0 (3.5), 10.2 (3.9), and 4.6 (1.6), respectively. In the patients, the proapo A-I concentration was positively correlated with the concentration of high-density lipoprotein subtype 2 cholesterol (HDL2-C) (r = 0.736), and the proapo A-I/total apo A-I ratio was correlated inversely with the HDL3-C concentration (r = -0.609). The activity of proapo A-I converting enzyme in patients with liver cirrhosis (62 +/- 30 nmol/h per liter) was significantly (P < 0.01) lower than that in normal subjects (172 +/- 55 nmol/h per liter). The increases of the plasma proapo A-I concentration and ratio in patients with liver cirrhosis may be caused by a decreased production of the converting enzyme in the liver. The increase of plasma proapo A-I may thus also affect the circulating HDL subtypes. PMID- 8419060 TI - Rapid diagnosis of phenylketonuria by quantitative analysis for phenylalanine and tyrosine in neonatal blood spots by tandem mass spectrometry. AB - A new method for quantifying specific amino acids in small volumes of plasma and whole blood has been developed. Based on isotope-dilution tandem mass spectrometry, the method takes only a few minutes to perform and requires minimal sample preparation. The accurate assay of both phenylalanine and tyrosine in dried blood spots used for neonatal screening for phenylketonuria in North Carolina successfully differentiated infants who had been classified as normal, affected, and falsely positive by current fluorometric methods. Because the mass spectrometric method also recognizes other aminoacidemias simultaneously and is capable of automation, it represents a useful development toward a broad-spectrum neonatal screening method. PMID- 8419061 TI - Home testing: to do or not to do? PMID- 8419062 TI - Differences in serum potassium concentrations in normal men in different geographic locations. AB - Hypokalemia has been associated with the taking of gossypol, a potential oral antifertility drug for men. Because the frequency of this response differed in different parts of the world, this study was done to learn if "normal" serum [K+] also differed. [K+] was measured by flame photometry in serum from apparently normal men from Austria (n = 30), China (53), Brazil (100), the Dominican Republic (38), and the US (103), and in plasma from Nigerian men (82). The mean (SD) for [K+] in Chinese men, 3.82 (0.27) mmol/L, was lower than that in Brazilians [4.06 (0.29) mmol/L], Austrians [4.14 (0.44) mmol/L], Dominicans [4.37 (0.33) mmol/L], or Americans [4.38 (0.37) mmol/L]. Apparently there are regional differences in average serum [K+], with men in China having lower serum [K+] than men elsewhere. This may predispose them to hypokalemia. PMID- 8419063 TI - Proficiency test performance as a predictor of accuracy of routine patient testing for theophylline. AB - Proficiency testing (PT) is pivotal in assessing laboratory qualifications for certification and licensure. PT is expected to typify routine assay performance and determine whether the laboratory is producing clinically useful test results. Conventional schemes use mail-distributed test specimens and are often criticized as measuring the best possible laboratory performance, principally because of special practices associated with processing PT specimens. We used on-site proficiency tests and split samples to evaluate the ability of conventional PT schemes to accurately characterize routine laboratory performance. Using 412 assays of theophylline, performed routinely by 200 laboratories and subsequently in a reference laboratory, we found that the predictive value of PT performance in assessing quality of routine testing was high (100% for predicting substandard reliability of routine patient testing and 94% for excluding substandard reliability of patient testing). The imprecision of interlaboratory PT results was equivalent whether testing was observed (hand-carried specimens) or unobserved (mail-distributed specimens). Many methods used for determining theophylline concentration in serum were highly automated, closed, and precise analytical systems. The performance characteristics of these analytical systems are not easily manipulated by the analyst for purposes of improving PT outcome, and PT by use of mail-distributed test specimens is effective for assessing intralaboratory performance. PMID- 8419064 TI - HPLC of folinic acid diastereoisomers and 5-methyltetrahydrofolate in plasma. AB - We present a rapid, sensitive, and automated HPLC method with direct resolution of l-folinic acid (l-FA), d-folinic acid (d-FA), and 5-methyltetrahydrofolate (5MTHF) from plasma samples. Plasma (500 microL) is first extracted on solid phase (RP-18 cartridge). The dI-FA peak is collected on-line from a reversed phase column (RP-8, 119 x 2 mm, 4 microns: HPLC 1) and then automatically loaded onto a chiral stationary phase (human serum albumin, 150 x 4.6 mm, 7 microns: HPLC 2). The same mobile phase flows in both systems (0.2 mol/L Na2HPO4:1 propanol, 98:2, pH 6.2). HPLC 1 allows quantification of 5MTHF by absorption at 313 nm; HPLC 2, the quantification of l-FA and d-FA by electrochemical detection in the oxidation mode (total run time 18 min). Recoveries are > 80%. CVs for intra- and interassay reproducibilities are < 5% and 15%, respectively. Linearity of the response (0.1-1 mumol/L and 1-50 mumol/L, r = 0.99, P < 0.01) is satisfactory. The sensitivity limit is 50 nmol/L for 5MTHF and 20 nmol/L for l-FA and d-FA. This assay is substantially improved over existing methods regarding feasibility and is being used in pharmacokinetic investigations in cancer patients. PMID- 8419065 TI - Breath acetone analyzer: diagnostic tool to monitor dietary fat loss. AB - Acetone, a metabolite of fat catabolism, is produced in excessive amounts in subjects on restricted-calorie weight-loss programs. Breath acetone measurements are useful as a motivational tool during dieting and for monitoring the effectiveness of weight-loss programs. We have developed a simple, easy-to-read method that quantifies the amount of acetone in a defined volume of exhaled breath after trapping the sample in a gas-analyzer column. The concentration of acetone, as measured by the length of a blue color zone in the analyzer column, correlates with results obtained by gas chromatography. Using the breath acetone analyzer to quantify breath acetone concentrations of dieting subjects, we established a correlation between breath acetone concentration and rate of fat loss (slope 52.2 nmol/L per gram per day, intercept 15.3 nmol/L, n = 78, r = 0.81). We also discussed the possibility of using breath acetone in diabetes management. PMID- 8419066 TI - Limitations of quality control in physicians' offices and other decentralized testing situations: the challenge to develop new methods of test validation. AB - Traditional quality-control methods are effective for detecting systematic error caused by deterioration of reagents or instruments, but ineffective for detecting sporadic error, which is more likely to occur in low-volume testing environments. Decentralized testing performed by individuals without formal laboratory training has a high potential for sporadic errors. New methods for validating test results, used each time a test result is produced, should replace current quality control procedures. Under the rules of CLIA '88, manufacturers and the US Food and Drug Administration have an opportunity to develop new approaches to test validation. PMID- 8419067 TI - Quantification of beta 2-microglobulin and albumin in plasma and peritoneal dialysis fluid by a noncompetitive immunoenzymometric assay. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for measuring beta 2 microglobulin (beta 2m) and albumin in continuous ambulatory peritoneal dialysis (CAPD) fluid. Plasma concentrations of beta 2m were twofold greater in hemodialysis patients (41.3 +/- 13.3 mg/L) than in CAPD patients (23.6 +/- 5.5 mg/L) matched for duration of treatment. Measurement of beta 2m in CAPD fluid showed a substantial loss of this protein, approximately 31% of total body beta 2m, compared with a 5% loss of a protein of middle molecular mass (albumin). Because of the molecular sieving effects of the peritoneal membrane, peritoneal clearance of beta 2m was sixfold greater than that of albumin. Whether beta 2m losses prevent or delay the incidence of dialysis-induced amyloidosis in these patients remains to be established. PMID- 8419068 TI - Determination of metanephrines in plasma by liquid chromatography with electrochemical detection. AB - Metanephrines are O-methylated metabolites of catecholamines. We report the use of liquid chromatography with electrochemical detection to determine plasma concentrations of normetanephrine (NMN) and metanephrine (MN). Plasma NMN and MN in 32 normal volunteers and inpatients were compared with concentrations in 23 patients with pheochromocytoma. Metanephrines were adsorbed from plasma onto a cation-exchange column and eluted with ammoniacal methanol. The dried residue was dissolved in mobile phase and injected onto a reversed-phase column. Recoveries of NMN and MN from 1 mL of plasma averaged 50-70%, and results varied linearly with quantity injected over a range of 0.13-55 pmol. The detection limit was 25 fmol for NMN and 50 fmol for MN. Intra-assay CVs were < 5%. In normal volunteers and inpatients, plasma concentrations of NMN ranged between 0.12 and 0.73 nmol/L (mean 0.38 nmol/L), and MN between 0.06 and 0.63 nmol/L (mean 0.19 nmol/L). Plasma NMN concentrations were increased in all 23 patients with pheochromocytoma (range 1-172 nmol/L), whereas MN concentrations (range 0.10-382 nmol/L) were increased in only 9 patients. The assay method is reliable and sensitive and offers an approach to examine the extraneuronal metabolism of catecholamines. The method may also be useful in the diagnosis of pheochromocytoma. PMID- 8419069 TI - ANCA and associated diseases: immunodiagnostic and pathogenetic aspects. AB - The past decade has seen an explosion of data on the new group of autoantibodies known collectively as ANCA (anti-neutrophil cytoplasmic antibodies). ANCA are specific for granule proteins of granulocytes and monocytes and induce distinct fluorescence patterns, e.g. the cytoplasmic (classic) cANCA and the perinuclear pANCA. cANCA is induced by antibodies directed against Proteinase 3 (PR3; PR3 ANCA) in about 90% of all ANCA-positive sera, and pANCA is induced by antibodies against myeloperoxidase (MPO; MPO-ANCA) in about 40%. A further staining pattern, which does not have a clear cut association with a distinct granule protein, is sometimes seen in chronic inflammatory bowel diseases. PR3-ANCA are serological markers for Wegener's granulomatosis (WG) and MPO-ANCA are associated with certain subtypes of primary vasculitides. Evidence exists that both the autoantigen and ANCA participate in the pathogenesis of at least the group of 'ANCA-associated vasculitides'. PMID- 8419070 TI - Identification of DNA-binding proteins on human umbilical vein endothelial cell plasma membrane. AB - The binding of anti-DNA antibodies to the endothelial cell is mediated through DNA, which forms a bridge between the immunoglobulin and the plasma membrane. We have shown that 32P-labelled DNA bound to the plasma membrane of human umbilical vein endothelial cells (HUVEC) by a saturable process, which could be competitively inhibited by non-radiolabelled DNA. In addition, DNA-binding was enhanced in HUVEC that had been treated with IL-1 alpha or tumour necrosis factor alpha (TNF-alpha). DNA-binding proteins of mol. wt 46,000, 92,000, and 84,000 were identified by the binding of 32P-labelled DNA to plasma membrane proteins separated on SDS-PAGE. DNA-binding proteins of mol. wt 46,000 and 84,000 were also present in the cytosol and nucleus. Murine anti-DNA MoAb410 bound to a single band, at mol. wt 46,000, of plasma membrane protein, in the presence of DNA. Our results showed that DNA-binding proteins are present in different cellular fractions of endothelial cells. DNA-binding proteins on the cell membrane could participate in the in situ formation of immune deposits; and their presence in the cell nucleus suggests a potential role in the modulation of cell function. PMID- 8419071 TI - Specific increases in urinary excretion of anti-DNA antibodies in lupus mice induced by lysozyme administration: further evidence for DNA-anti-DNA immune complexes in the pathogenesis of nephritis. AB - We previously reported that lysozyme electrostatically inhibits the fibronectin mediated DNA binding to the glomerular basement membrane (GBM) and reduces in situ DNA-anti-DNA complex formation in the GBM in NZB/W F1 mice [1]. In this study, we further noticed significant increases in urinary excretion of anti-DNA antibodies and immune complexes (IC) in lysozyme-treated NZB/W F1 mice. Their clearance ratios of IgG anti-DNA antibody to whole IgG were markedly high compared with those of saline-treated animals. A large number of IgG and C3 positive granules were observed in the tubular cells of NZB/W F1 mice treated with lysozyme. On the contrary, nil or only small amounts of anti-DNA antibodies were detected in the urine of NZB/W F1 mice without lysozyme administration despite a large amount of proteinuria, suggesting entrapment of the antibodies in lupus glomeruli. Lysozyme neither inhibited the binding of anti-DNA antibodies to DNA or heparan sulphate nor did it displace anti-DNA antibodies and IC from the kidney homogenates of lupus mice. It thus appears that the inhibition of DNA binding to the GBM due to lysozyme reduced the entrapment of anti-DNA antibodies in the GBM, resulting in urinary excretion of the antibodies. PMID- 8419072 TI - Antiretroviral treatment reverses HIV-induced reduction in the expression of surface antigens on alveolar macrophages in AIDS patients. AB - MoAbs and immunoperoxidase methods were used to identify antigen-presenting and phagocytic cells and to assess expression of HLA-DR molecules on cells obtained by bronchoalveolar lavage (BAL) from 33 AIDS patients and nine normal volunteers. In 17 patients, not receiving antiretroviral therapy, the expression of HLA-DR molecules (MoAb RFDR1) as well as the percentages of cells expressing RFD1 marker for antigen-presenting cells and RFD7 marker for mature phagocytes were significantly reduced. However, in BAL obtained after commencing treatment with zidovudine (AZT) in 21 patients or with 2',3'-dideoxyinosine (DDI) in five patients, the expression of the markers studied was found to have returned to levels of expression seen in normal lavages. The changes observed were clearly associated with antiretroviral treatment and did not correlate with applications of other drugs, blood CD4 counts or presence of infectious organisms in BAL fluid. As the alterations in the expression of HLA-DR molecules and RFD1 marker on macrophages have been shown to be associated with functional capacities of these cells, the reversal of impaired expression of phenotypic markers on alveolar macrophages in AIDS patients by AZT and DDI signifies an important ability of these drugs to modify immune reactivity and emphasizes the need to monitor such functions in HIV disease. PMID- 8419073 TI - Nephrotoxic serum nephritis in nude rats: the roles of host immune reactions in the accelerated type. AB - By immunization with rabbit immunoglobulins and the injection of a subnephritogenic dose of rabbit nephrotoxic serum (NTS), accelerated-type nephrotoxic serum nephritis (NTN) was induced in heterozygous (rnu/+) rats but not in athymic nude (rnu/rnu) rats. By transferring rat antibody against rabbit immunoglobulins, marked proteinuria was induced also in nude rats (202.0 +/- 98.4 mg/day on day 3) as in rnu/+ rats (122.6 +/- 35.3 mg/day on day 3). No marked differences in histological findings could be found between both groups. The most marked increase in the number of intraglomerular infiltrating cells was observed in heterozygous rats indicating that the presence of thymus-derived cells leads to the accumulation of more cells in glomeruli. We conclude that humoral immunity alone is enough to accelerate the pathogenic mechanism which induces glomerular injury with heavy proteinuria in this model. PMID- 8419074 TI - Effect of chlorpromazine on kinetics of injected monoclonal antibody in MoAb induced glomerular injury. AB - The effect of chlorpromazine, one of several calmodulin antagonists that inhibit cytoskeletal movement, on the local kinetics of injected proteinuria-inducing MoAb 5-1-6 was examined to test the hypothesis that proteinuria is inhibited if the antigen recognized by MoAb 5-1-6 or injected MoAb remains on the surface of epithelial foot processes. MoAb 5-1-6 was injected into both chlorpromazine treated (5 mg/100 g body weight) and untreated rats. As a positive control for the chlorpromazine treatment, anti-Fx1A serum was also injected into other chlorpromazine-treated and untreated rats. Chlorpromazine inhibited neither the change in localization of injected MoAb 5-1-6 nor proteinuria, although it showed an inhibitory effect on redistribution of immune complex and the fixation of complement in passive Heymann glomerulonephritis induced by injection of anti Fx1A serum. We conclude that the kinetics of bound MoAb 5-1-6 are regulated by a system different from that operating in passive Heymann glomerulonephritis. PMID- 8419075 TI - Modulation of CD4 by suramin. AB - Suramin is a polysulphonated compound which can selectively bind to, and inhibit the activity of, a wide range of growth factors. There has been renewed interest recently in suramin as an anti-cancer agent and therefore we have studied its effects on lymphocyte subset populations and recombinant human IL-2 (rhIL-2) activation on lymphocytes in vitro. In the presence of rhIL-2 (1000 U/ml), suramin (200 micrograms/ml) caused a decrease in percentage of cells expressing the predominantly T cell antigen CD3; no change in percentage of cells expressing the T suppressor/cytotoxic subset antigen, CD8; a small rise in those expressing the natural killer cell antigen, CD56; and a large significant fall in those expressing the T helper subset antigen CD4 (48.51% versus 27.97%; P < 0.001). CD4 modulation by suramin was also found on the CD4+ cell lines CEM and MOLT-4. The effect of suramin on rhIL-2-induced activation antigen expression remains equivocal, since a small rise in CD25 expression and small falls in CD71 and HLA Dr expression were recorded. The modulatory effect of suramin on CD4 expression was not reversible over a 96-h culture period in its continued presence. However, on removal of suramin by extensive washing, recovery of CD4 expression was detected within 24 h. Suramin-induced modulation, but not PMA-induced modulation, could be partially inhibited by preincubation with tyrphostin (12 microM), a tyrosine kinase inhibitor. PMID- 8419076 TI - No restriction of intrathyroidal T cell receptor V alpha families in the thyroid of Graves' disease. AB - Recently it has been reported that the intrathyroidal T cells in Graves' disease display restriction in V alpha T cell receptor (TcR) gene family usage, although this is not found with TcR V beta gene families in the same individuals. We have performed a qualitative analysis of TcR V alpha family usage in 12 patients with Graves' disease by reverse transcription and polymerase chain reaction (PCR) amplification of RNA extracted from isolated, unstimulated intrathyroidal lymphocytes and from snap-frozen whole thyroid specimens. No restriction was observed, with 10-15 V alpha gene families being amplified in all cases. The pattern of usage was similar to that in peripheral blood lymphocytes derived from normal subjects (n = 3) and from patients with Graves' disease (n = 3), as well as that present in the thyroids of patients with non-autoimmune toxic multinodular goitre (n = 4). These results indicated that there is no marked restriction of the unselected intrathyroidal T cell population in patients with Graves' disease who have been treated with antithyroid drugs. PMID- 8419077 TI - Asp57-negative HLA DQ beta chain and DQA1*0501 allele are essential for the onset of DQw2-positive and DQw2-negative coeliac disease. AB - The genetic predisposition to coeliac disease is associated with the HLA DQw2 allele. Coeliac patients lacking the DQw2 allele are very rare and always exhibit the DR4-DQw3 haplotype. We performed oligotyping of polymerase chain reaction (PCR)-amplified DQA1 and DQB1 genes in six DQw2-negative and 30 DQw2-positive coeliac patients. The DQB analysis showed that all six DQw2-negative patients possessed the DQB1*0302 allele. The other DQB alleles found in five of these patients were DQB1*0501, DQB1*0604 and DQB1*0302. The DQ beta chains encoded from all these alleles have the replacement of aspartic acid residue at position 57 (Asp57), as well as the DQB1*0201 allele which was found in all 30 DQw2-positive coeliac patients. The DQw2-negative proband who lacked the homozygous Asp57 replacement exhibited the DQA1*0501 allele in the DQA1 gene. The DQA1*0501 allele was also found in 27 of the 30 DQw2-positive coeliac patients. Among this group of coeliacs, the four cases lacking the DQA1*0501 allele exhibited the homozygous Asp57 replacement in the DQ beta chain. Our results indicate that Asp57-negative DQ beta alleles are involved in both DQw2-positive and -negative coeliac patients. Moreover, when the Asp57-negative DQ beta chain is encoded from only one of the two DQB1 genes the DQA1*0501 allele is always present. PMID- 8419078 TI - Extracellular acidic pH modulates oxygen-dependent cytotoxic responses mediated by polymorphonuclear leucocytes and monocytes. AB - In the present study, we compared the ability of human neutrophils and monocytes to display oxygen-dependent cytotoxic responses at pH 7.4 and 6.2. Our results show that cytotoxicity induced by immune complexes (IC), zymosan, N-formyl methionyl-leucyl-phenylalanine (FMLP) and concanavalin A (Con A) were markedly increased when they were carried out at pH 6.2 instead of pH 7.4. Cytotoxicity induced by phorbol myristate acetate (PMA), on the contrary, was significantly decreased at pH 6.2. It is noteworthy that cytotoxic responses induced by IC, zymosan and Con A were also increased when, 2 h after effector cell stimulation at pH 6.2, cytotoxicity was measured at pH 7.4. Finally, when we examined possible mechanisms involved in the augmentation of cytotoxicity, we observed that the oxidative response of IC-stimulated neutrophils, measured as chemiluminescence emission, was not increased at pH 6.2, on the contrary, it was significantly decreased. The relevance of these results is discussed. PMID- 8419079 TI - Interferon-gamma activates the oxidative killing of Candida albicans by human granulocytes. AB - Although granulocytes are essential for the resistance against infections with Candida albicans, these cells do not kill the ingested yeast optimally. Various cytokines can enhance functional activities of granulocytes, but until now only interferon-gamma (IFN-gamma) has been applied more widely, namely in patients with chronic granulomatous disease. Since it is not certain whether IFN-gamma is able to enhance the candidacidal activity of granulocytes the present study was undertaken. Human granulocytes incubated with various concentrations of recombinant human IFN-gamma (rIFN-gamma) were studied for the phagocytosis and intracellular killing of C. albicans and their oxygen metabolism after stimulation with opsonized Candida. Results showed a small increase in the rate of phagocytosis and a dose-dependent increase of the intracellular killing of C. albicans and the production of H2O2. The increased candidacidal activity and H2O2 production by rIFN-gamma-stimulated granulocytes were inhibited by diphenylene iodonium (DPI). From these results it is concluded that the increased candidacidal activity of granulocytes activated by rIFN-gamma is caused by the increased production of reactive oxygen radicals. PMID- 8419080 TI - Mononuclear phagocytes from human bone marrow progenitor cells; morphology, surface phenotype, and functional properties of resting and activated cells. AB - After 3-4 weeks culture of human bone marrow cells in medium supplemented with IL 3, macrophage- (M-CSF), and granulocyte-macrophage colony-stimulating factor (GM CSF), the firmly adherent cells exhibited the morphologic features of mononuclear phagocytes and were strongly esterase-positive. Flow cytometric analysis revealed a rather homogeneous cell population with marked autofluorescence; the large majority of the cells expressed CD14, CD11a, b, and c, Fc receptors for IgG, Fc gamma RI, II, and III, and HLA class II molecules. Interferon-gamma (IFN-gamma), bacteria, and bacterial products modulated expression of some of the surface markers, induced and/or enhanced respiratory burst, phagocytic activity, secretion of tumour necrosis factor, and tumouricidal activity; in contrast, these cells were not able to generate reactive nitrogen intermediates. PMID- 8419081 TI - Interaction of papain-digested HLA class I molecules with human alloreactive cytotoxic T lymphocytes (CTL). AB - Acute immunological rejection events of transplanted allogeneic organs are strongly dependent on T cell reactivity against foreign MHC products. The recognition requirements of alloreactive cytotoxic T cells are of particular interest for finding approaches to modulating alloreactivity. The role of the allogeneic MHC molecule itself and/or an associated peptide in the interaction with the T cell receptor is still, however, unclear. Our studies have focused on the interactions of papain-digested HLA class I molecules with alloreactive CD8+ CTL. These polypeptides, consisting of the polymorphic alpha 1 and alpha 2 and the monomorphic alpha 3 domains, were used in both soluble and immobilized form to study their functional effects on anti-HLA-A2 reactive CTL. Purified polypeptides were of molecular mass 32-34 kD. HLA-A2 polypeptides (0.55 micrograms/ml) in soluble form induced half-maximal reduction of CTL cytotoxicity. These concentrations were quantitatively comparable to the effective doses of intact HLA class I molecules, which contain the hydrophobic transmembrane domain and the intracytoplasmic tail. In addition, specific activation requirements of these CTL were investigated in a serine esterase release assay. Maximal degranulation was observed after 2 h of antigen contact. Purified HLA class I molecules allospecifically activated the anti-HLA-A2 CTL to degranulate serine esterase, when immobilized on plastic microtitre plates. Thus, polypeptides containing the polymorphic alpha 1 and alpha 2 domains of human class I molecules potentially modulate the cytotoxic T cell response. This might have implications for the reduction or prevention of allograft rejection in recipients of foreign organs. PMID- 8419082 TI - Immunoglobulin G subclass responses to mycobacterial lipoarabinomannan in HIV infected and non-infected patients with tuberculosis. AB - Immunoglobulin G subclass responses to lipoarabinomannan (LAM) of Mycobacterium tuberculosis were determined by ELISA in both HIV-1 antibody positive (n = 31) and negative (n = 43) patients with tuberculosis (TB). Responses were also studied in a group of healthy controls (n = 16) and HIV-1 antibody positive (n = 60) individuals without TB. IgG2 antibodies were the predominant subclass, being present in 25 of 43 non-HIV-infected TB patients (58%) and in 11 of 31 HIV infected TB patients (35%). However, HIV+ TB patients also showed IgG4 (n = 16; 52%), and IgG1 (n = 4, 13%) responses to LAM, whereas these subclasses were absent in sera from HIV-TB patients. Individuals in both non-tuberculous control groups showed no antibody responses to LAM. The influence of HIV infection on B cell responses to LAM, and possible mechanisms for antibody-mediated regulation of immunity to TB, are explored. PMID- 8419084 TI - Raised levels of tumour necrosis factor-alpha and neopterin, but not interferon alpha, in serum of HIV-1-infected patients from Ethiopia. AB - Serum levels of tumour necrosis factor-alpha (TNF-alpha), neopterin and interferon-alpha (IFN-alpha) were determined by immunoradiometric assays in 60 HIV-1+ and 20 HIV-1- subjects from Ethiopia. Swedish samples were used as reference material. The Ethiopian HIV-1+ subjects were found to have significantly increased TNF-alpha and neopterin, but not IFN-alpha levels. Increased levels of TNF-alpha and neopterin were frequently found in Ethiopian asymptomatic subjects (37% and 47%), and the concentration increased in patients with AIDS (83% and 90% respectively). The levels of the two substances and the proportion of patients with higher TNF-alpha values were lower in the corresponding Swedish subjects. The proportion of sera with raised levels of IFN alpha was very low (asymptomatic 4%, and AIDS 7%) in Ethiopian subjects. These results suggest a very early increase in the TNF-alpha production and activation of the cellular immune response, and a low level of IFN-alpha synthesis in the natural course of HIV infection in Ethiopia. The aberrations may contribute to a rapid progress of immunodeficiency and cachexia often seen in Ethiopian patients. PMID- 8419083 TI - Differences in cytokine secretion by intestinal mononuclear cells, peripheral blood monocytes and alveolar macrophages from HIV-infected patients. AB - Mononuclear cells of the lamina propria (LpMNC), isolated from endoscopically taken biopsies of the large bowel from AIDS patients, were analysed for their ability to secrete tumour necrosis factor-alpha (TNF-alpha), IL-1 beta and IL-6. Stimulation of LpMNC from normal controls with pokeweed mitogen (PWM) led to a time- and dose-dependent enhancement of TNF-alpha, IL-1 beta and IL-6 secretion. In contrast, PWM stimulation of LpMNC from AIDS patients resulted in only a small increase in TNF-alpha release. Constitutive secretion of IL-1 beta and IL-6 in these patients was already increased to the concentration range of stimulated cells from normal controls and could not be further increased, probably due to maximal in vivo stimulation. Secretion of TNF-alpha, IL-1 beta and IL-6 by peripheral blood monocytes (PBM) and alveolar macrophages from AIDS patients was elevated with or without stimulation compared with normal controls. Obviously, the regulation of TNF-alpha secretion is dependent on the microenvironment. Since it is known that interferon-gamma (IFN-gamma) may induce the production of TNF alpha, the secretion of this cytokine was examined. Release of IFN-gamma was constitutively and under stimulation lowered in LpMNC from AIDS patients compared with normal controls. Addition of IFN-gamma to LpMNC did not result in enhanced TNF-alpha secretion. Our data indicate a defective function of intestinal mononuclear cells in AIDS patients as shown by the diminished TNF-alpha secretion. PMID- 8419085 TI - Relationship of in vitro phagocytosis of serotype 14 Streptococcus pneumoniae to specific class and IgG subclass antibody levels in healthy adults. AB - The role of specific IgG2 antibody in the protection against serious infection with Streptococcus pneumoniae is unclear. We therefore decided to investigate the relationship between serum antibody levels and opsonization and phagocytosis of this microorganism. We have measured serum IgM, IgA and IgG subclass antibody specific for pneumococcal capsular polysaccharide and in vitro phagocytosis of serotype 14 pneumococcus by polymorphs, in healthy adults before and after immunization with Pneumovax II. IgM and IgG2 were the predominant anti pneumococcal antibodies seen, IgA and IgG1 being present at low titre. No significant relationship of phagocytosis with specific IgM and IgA antibodies was found. However, both specific IgG1 and IgG2 antibodies in post-immunization sera correlated significantly with phagocytosis of the pneumococcus in the presence of complement (r = 0.57, P = 0.029 and r = 0.59, P = 0.022 respectively). After heat inactivation, the remaining opsonic activity of sera correlated only with levels of specific IgG2 antibody (r = 0.61, P = 0.0006). Whereas phagocytosis supported by specific IgG1 and IgG2 antibody to serotype 14 pneumococcus after immunization is mediated by complement activation, IgG2-specific antibody in high titre may also be able to function by complement-independent interaction with Fc gamma receptors on polymorphs. PMID- 8419086 TI - Mycobacterial 65-kD heat shock protein induces release of proinflammatory cytokines from human monocytic cells. AB - Monocytes having phagocytosed mycobacteria are known to present the bacterial 65 kD heat shock protein (hsp) on their cell surface to alpha beta and gamma delta T lymphocytes. Cytotoxic CD4+ cells may then lyse monocytes expressing mycobacterial 65-kD hsp. However, it is not known whether 65-kD hsp directly stimulates monocyte functions other than antigen presentation. This study has demonstrated that following extraction of bacterial lipopolysaccharide, purified recombinant mycobacterial 65-kD hsp may directly activate THP-1 cells, a human monocytic line, to accumulate mRNA for and secrete tumour necrosis factor (TNF), a cytokine important in granuloma formation, the characteristic host immune response to mycobacterial infection. TNF gene expression and secretion following stimulation by hsp was dose-dependent and abolished by heat-induced proteolysis. Subsequently, THP-1 cells secreted IL-6 and IL-8, cytokines involved in recruitment and differentiation of T lymphocytes. The data indicate that secretion of proinflammatory cytokines from monocytes activated by mycobacterial 65-kD hsp may be important in the host immune response and in the development of antigen-specific T cell-mediated immunity. PMID- 8419087 TI - Quantification of antibody-secreting lymphocytes that react with Pf155/RESA from Plasmodium falciparum: an ELISPOT assay for field studies. AB - We have adapted the enzyme-linked immunospot assay (ELISPOT) to enumerate the cells from Plasmodium falciparum-primed donors that produce IgG in vitro in response to malaria antigens. In vitro activation of cell cultures with two synthetic peptides (EENVEHDA)4, and (LGRSGGDIIKMQTL) corresponding to immunodominant T cell epitopes of the ring-infected erythrocyte surface antigen (Pf155/RESA) gave specific antibody-secreting cells (ASC) in five and six of the 15 P. falciparum-primed donors from Cameroon. Antibodies produced after a stimulation by synthetic peptides reacted also with total parasite proteins. However, crude P. falciparum antigen did not trigger a higher number of cells than did synthetic peptides. The absence of significant relation between the presence of sera antibodies and in vitro ASC against the same peptide suggests that the kinetics of circulating primed lymphocytes and antibodies are different. We evaluated 0.04-0.29% of peripheral blood B cells to be the frequency of memory cells specific to a single Pf155/RESA epitope in these donors. This study suggests that the ELISPOT assay should permit the analysis of B cell responses to malarial antigens at the single-cell level and its applicability to epidemiological field studies. This assay should be well suited to the identification of T helper epitopes capable of inducing the production of antibodies by human B cells, and will constitute an important tool for the selection of immunogens to be included in a subunit vaccine. PMID- 8419088 TI - Antigen-dependent in vitro culture of protective T cells from BCG-primed mice. AB - Induction of protective immunity against pathogenic mycobacteria depends on vaccination with live organisms such as Bacille Calmette-Guerin (BCG). However, it is not known how many and which antigens are involved in the protective host response. In this study, we developed a system of antigen-dependent in vitro culture which is suitable for the analysis of protective subunits, presented in a soluble form. Spleen cells from Mycobacterium bovis BCG-immune mice, enriched for T cells and depleted of adherent cells on a column of G-10 Sephadex, were cultured for periods varying between 3 and 14 days before transfer and challenge with M. tuberculosis in irradiated hosts. Following 10 days in culture, immune T cells sustained their capacity to transfer protection to tuberculous infection when incubated in the presence of either live BCG or a soluble extract from M. tuberculosis, but lost this ability when cultured in the absence of antigen, or in the presence of the polyclonal mitogen concanavalin A. One immunodominant antigen, represented by the recombinant 38-kD antigen, failed to sustain the adoptive protection, despite pronounced stimulation of lymphoproliferation in culture. Antigenic in vitro stimulation of protective T cells was accompanied by enhanced responsiveness to exogenous IL-2. The experimental system described may be generally suitable to test in vitro the protective potentials of soluble molecular subunits of mycobacteria. PMID- 8419091 TI - The nursing course guide: asset or trap? PMID- 8419089 TI - T cell redistribution kinetics after secondary infection of BALB/c mice with respiratory syncytial virus. AB - BALB/c mice were infected intranasally with live respiratory syncytial virus (RSV) and reinfected 4 weeks later. At regular intervals thereafter groups of animals were killed and T cell subsets were determined in blood, spleen and bronchoalveolar lavage (BAL) with flow cytometry employing T cell subset-specific MoAbs. Total lymphocyte counts in the peripheral blood decreased 1-3 days after infection, returning to preinfection levels on day 8 (P = 0.0111). Simultaneously, a marked increase of lymphocytes was noted in the BAL, reaching a maximum at day 8 (P < 0.0001). Both CD4+ and CD8+ T cells decreased in the blood on day 1-3 (P < 0.0097 and P = 0.003 respectively), and increased in the BAL progressively towards a maximum at day 8 (P < 0.0001). In BAL, CD4+ cells increased 35-fold and CD8+ cells 27-fold during the first week after reinfection. On the other hand, in the spleen a significant decline of CD4+ and CD8+ cells was noted 1 day post-infection (P = 0.0002). It is concluded that a strong T cell redistribution response among systemic and mucosal tissues occurs after reinfection with RSV. The kinetics of this response differ both quantitatively and qualitatively from the T cell response after primary infection. The magnitude of cell traffic is more pronounced in blood, spleen and BAL than after primary infection. CD4+ T cells are more intensively distributed to the lungs than after primary infection. PMID- 8419090 TI - T cells are responsible for the enhanced synovial cellular immune response to triggering antigen in reactive arthritis. AB - In reactive arthritis (ReA) there is specific proliferation of synovial fluid (SF) mononuclear cells (MNC) to the triggering bacterial antigen; comparatively little or no response is seen in peripheral blood (PB). To investigate the mechanism of this elevated local immune response, we examined patients with typical ReA who showed an enhanced antigen-specific synovial immune response in bulk culture. Using separated fractions of T cells and antigen-presenting cells (APC) from PB and SF we showed that the synovial T cells rather than SF APC are responsible for the specific proliferation. By limiting dilution analysis, the frequency of T cells responding to the specific antigen was found to be significantly increased compared with the frequency of irrelevant antigen specific T cells. Furthermore, the frequency of T cells responding to the specific antigen was higher in SF (between 1/619 and 1/4846, mean 1/2389) than in PB (between 1/1286 and 1/16,279, mean 1/7350). We conclude that the specific synovial cellular immune response in ReA is mainly due to an expansion of antigen specific T cells within the joint. However, the non-specific hyper-reactivity of SF T cells and differences between SF and PB APC may make a more minor contribution. PMID- 8419092 TI - Association of reported infant crying and maternal parenting stress. AB - Infant crying is a well-recognized source of parental concern and anxiety, but little is known about other possible effects of excessive crying on parents or caretakers. The objective of this descriptive study was to investigate the relationship between reported infant crying and parenting stress. Mothers, who were consecutively enrolled at the time of their infants' well-child checkups, reported the number of minutes their infant generally cried in a 24-hour day. Excessive crying was defined as more than three hours per 24-hour day. Seventy five of 94 mothers subsequently completed the Parenting Stress Index (PSI) at their infant's 4- or 6-month checkup. Mothers who reported excessive crying were 5.7 times more likely to score high on the reinforcement subscale of the PSI. Although the direction of this association is not known, it indicates that mothers who report excessive crying are more likely than other mothers to perceive a lack of positive reinforcement from their infants. PMID- 8419093 TI - Perceptions of vaccine efficacy, illness, and health among inner-city parents. AB - A resurgence of measles in the past decade has focused attention on the limitations of current immunization programs, particularly for inner-city, low income populations. As part of a larger study of immunization rates, we discussed perceptions of disease severity and vaccine efficacy, as well as the prioritization of the tasks of parenthood, with 40 parents of infants living in inner-city Baltimore to discover their beliefs about immunization. Vaccines were considered only partly successful; susceptibility to chickenpox after vaccination was repeatedly cited as evidence of vaccine failure. Fever was seen as a primary indicator of illness; thus, vaccines were believed to cause, rather than prevent, illness. Immunization was not considered a high-priority parental responsibility. These findings suggest future interventions be aimed at changing parental perceptions of vaccines as ineffective and of fever after immunization as an indicator of illness. Finally, immunizations should be made easily available, even during clinic visits for a child's illness. PMID- 8419094 TI - Group D streptococcal bacteremia in children. A review of 72 cases in 12 years. AB - A review was conducted of 72 cases of pediatric group D streptococcal (GDS) bacteremia treated at our institution during a 12-year period. These 72 cases represented 90% of all instances in which this organism was isolated by blood culture (in eight others, GDS was considered a contaminant); the rate of isolation of this organism relative to all positive blood cultures during this time period was 1.3%. Infection was nosocomially acquired in 25 cases; 18 occurred in an intensive care unit. At the time their positive blood culture was obtained, 25 patients were afebrile and 10 patients were receiving parenteral antibiotic therapy to which the isolate exhibited in vitro susceptibility. In 31 cases, GDS was isolated by blood culture in conjunction with another organism, most frequently Staphylococcus epidermidis. Underlying medical conditions or foci of infection associated with GDS bacteremia were identified in 65 patients; the most common were the presence of an indwelling central venous catheter (23), a variety of lesions of the gastrointestinal tract (21), and pulmonary infiltrate (15). Bacteremia was associated with GDS meningitis in three patients who had had no prior neurosurgical procedure. The overall mortality rate was 20%; nearly two thirds of all deaths occurred in patients younger than 1 year of age. PMID- 8419095 TI - Propylthiouracil hepatotoxicity. A review and case presentation. AB - Propylthiouracil (PTU) is widely used to treat patients with hyperthyroidism. In rare cases, this drug has been found to have severe toxic effects on the liver. The case of a 14-year-old girl treated with PTU for hyperthyroidism who developed jaundice, severe hepatocellular dysfunction, and hepatomegaly is reported. Her condition gradually deteriorated, and she developed paranoid ideation, profound lethargy, and peripheral edema. After three weeks of prednisone therapy, clinical and laboratory signs of improvement were observed. This patient was one of only five pediatric cases among the 16 reported cases of PTU liver toxicity reported to date. Her history and the fatal outcome in some reported cases demonstrate the high degree of sensitivity required to recognize this potential complication in patients treated with PTU, particularly since its immediate discontinuance and steroid-therapy intervention may lead to recovery. PMID- 8419096 TI - Behavioral antecedents of pediatric poisonings. AB - To determine how the behavior of 3- to 7-year-olds might contribute to pediatric poisoning, a taxonomy of pediatric ingestions was conceptualized and tested. Thirty-nine of 50 caretakers who called the Middle Tennessee Poison Center about poison ingestions by a child in this age group provided details of the poisoning history and family characteristics. Histories were coded by a psychologist according to the 14 antecedent conditions of the poisoning taxonomy. Improper storage, noncompliant behavior, curiosity, misinterpretation of the substance, improper child monitoring, and imitative behavior were judged the most common antecedent conditions of these ingestions. Lack of knowledge of poison prevention methods was infrequently coded. Noncompliant behavior as a condition of the poisoning was associated with parental reports of general child behavioral problems. Ingestions involving imitative behavior were associated with parental report of poor social supports. The findings highlight the need to address behavior in designing poison prevention programs. PMID- 8419097 TI - Feeding premature infants. AB - We have tried to offer a rational approach to the methods of premature infant feeding. Most of our information is incomplete, and many of the presumptions used in deciding feeding technique are not based on reliable scientific data. The calculations presented in this discussion are valuable as teaching methods in the nursery. The complexity of the questions about premature infant feeding makes it necessary for personnel in the neonatal unit to focus regularly on infant feeding and to allow for time dedicated to considering their suggestions. PMID- 8419098 TI - Newborn cardiac arrhythmias associated with maternal caffeine use during pregnancy. PMID- 8419099 TI - Simultaneous Kawasaki disease and group A streptococcal pharyngitis. PMID- 8419100 TI - Improved lactation with metoclopramide. A case report. PMID- 8419101 TI - Indomethacin and cation-exchange resin in the management of pseudohypoaldosteronism. PMID- 8419102 TI - What, me worry? A survey of adolescents' concerns. AB - Adolescence is generally regarded as a time of transition demanding considerable adaptation. Failing to successfully negotiate the developmental hurdles associated with this period can have serious physical and psychosocial consequences (e.g., teenage pregnancy, suicide, disruption of social relationships, poor school performance). Efforts to identify problems that adolescents typically experience have been useful in facilitating a better understanding of this developmental phase. However, the perceived concerns or worries of adolescents themselves have been neglected as an additional source of useful information. This study assessed the worries of 622 adolescents between the ages of 12 and 20 years on an 80-item self-report measure, the "Things That Worry Me" scale. Findings indicated consistent concerns related to terrorism, adolescents' self-esteem, parents' physical and mental health, and adolescents' dating and sexual relationships. Gender- and race-specific differences are discussed and implications for future research and clinical applications are offered. PMID- 8419103 TI - Unusual acute, nonallergic eruptions of the scalp from combined use of minoxidil and retinoic acid. PMID- 8419104 TI - A dermatologic diary. Portrait of a practice. PMID- 8419105 TI - Childhood vitiligo. PMID- 8419106 TI - Pustular psoriasis. AB - Pustular psoriasis may appear as localized pustular psoriasis, which runs a chronic course, or, in a more severe state, generalized pustular psoriasis. Precipitating factors influencing both local and generalized forms include various drugs (eg, lithium, hydrochloroquine), irritative topical therapy (eg, coal tar), dental and upper respiratory infections, pregnancy, and solar irradiation. Both adults and children are affected, but infantile and juvenile pustular psoriasis is rare. It is recommended that provoking factors be eliminated as the first line of management. Short-contact anthralin therapy and topical psoralen/ultraviolet A can be used in treating localized pustular psoriasis. Methotrexate, hydroxyurea, etretinate, dapsone, cyclosporin A, as well as systemic corticosteroids may be needed in life-threatening cases. PMID- 8419107 TI - Use of cotton-tipped applicators during skin examination and dermatologic surgery. AB - The cotton-tipped applicator is a widely available and versatile tool for the dermatologist. In addition to the common uses of this inexpensive tool there are many other ways for the dermatologist to use cotton-tipped applicators. From its usefulness in dermatologic examination and potassium hydroxide preparations to its multiple surgical applications, the cotton-tipped applicator is a valuable and underrated tool. PMID- 8419108 TI - Localized multiple neurilemmomas of the lower extremity. AB - A neurilemmoma usually presents as a solitary subcutaneous tumor adherent to a peripheral nerve. Multiple localized neurilemmomas may occur as an isolated cutaneous finding or as part of the syndrome of neurilemmomatosis. We report a case of multiple neurilemmomas appearing on an extremity after trauma. The multiplicity and localization of the tumors are rare and illustrate that neurilemmomas are not invariably solitary tumors and that their distribution can be regional. We review the previously reported cases of multiple neurilemmomas and review the characteristics that help the clinician to differentiate between multiple localized neurilemmomas and neurilemmomatosis. PMID- 8419109 TI - Phytophotodermatitis: a visit to Margaritaville. AB - Phytophotodermatitis is a phototoxic eruption following contact with photosensitizing compounds and long-wave ultraviolet light. The most common phototoxic compounds are the furocoumarins contained in a wide variety of plants, especially of the Umbelliferae, Rutaceae, and Moracea families. Commonly occurring photosensitizing plants include citrus fruits such as limes and oranges and many vegetables, notably celery, parsnip, parsley, carrots, and dill. PMID- 8419110 TI - Linear bacterial dissection. AB - An unusual case of an erythematous indurated arcuate plaque with a curvilinear appearance was noted after a patient manipulated an ingrown beard hair. During removal of a biopsy specimen, a purulent discharge was expressed that had evidently dissected through the dermis forming this interesting shape. This may be similar to a recently reported entity described by Shelley and Shelley as linear bacterial dissection. Culture of the discharge grew Staphylococcus aureus and Klebsiella pneumoniae. Complete resolution occurred after drainage and antibiotic therapy. Cellulitis and erysipelas typically do not show central clearing or form arcuate plaques. Aside from lymphangitis, thrombophlebitis, and sinus tracts of carbuncles, dermal infectious processes rarely present as linear or arcuate plaques. The differential diagnosis and a review of the literature are presented. PMID- 8419111 TI - Atypical fibroxanthoma of the cheek: a case report. AB - Atypical fibroxanthoma is a cutaneous soft tissue tumor that may mimic squamous cell carcinoma and melanoma on clinical and histologic examination. Immunohistochemical stains for cytokeratin, S100 protein, and vimentin may be helpful in differentiating atypical fibroxanthoma from the latter two diagnoses. We report a patient with atypical fibroxanthoma of the cheek who underwent successful local excision. The clinical and laboratory features of this neoplasm are reviewed. PMID- 8419112 TI - Fluoxetine treatment of trichotillomania and depression in a prepubertal child. AB - The case of a ten-year-old boy with a two-year history of trichotillomania and depression is presented. Imipramine was unsuccessful in treating trichotillomania and showed limited success in alleviating depression. Treatment with low-dosage fluoxetine (10 mg daily) led to marked improvement of both trichotillomania and depression. PMID- 8419113 TI - Ofloxacin treatment of difficult infections of the skin and skin structure. AB - We report an open evaluation of ofloxacin therapy, 400 mg every twelve hours (parenterally followed by oral treatment) in hospitalized subjects with infections of the skin and skin structure that were recalcitrant. There were 105 evaluable subjects with an average age of fifty-five years, thirty-two (30 percent) of whom had diabetes mellitus, and in sixty-one (58 percent), a regimen of parenteral antibiotics (typically combinations involving aminoglycosides) had recently failed. There were 115 pathogens isolated; the most common were Enterobacteriaceae (fifty-five), Staphylococcus aureus (thirty), coagulase negative Staphylococcus (ten), and Pseudomonas aeruginosa (ten). Overall, 103 (90 percent) pathogens were eradicated by therapy, and twelve (10 percent) persisted, including four coagulase-negative Staphylococcus which emerged as resistant during therapy. For thirty-five (33 percent) subjects, colonization or superinfection was documented; seven of these organisms (five Enterococcus, one S. aureus, and one P. aeruginosa) were resistant to treatment with ofloxacin. Clinical response was rated as cure in seventy (67 percent) subjects, improvement in twenty-one (20 percent) subjects, and failure in fourteen (13 percent). Failures were accompanied by persistence of the original pathogen (eight), persistent or resistant superinfection (six), or both (one). Adverse effects were infrequent, mild, and self-limiting. There was one death during the study, attributed to septic shock after postoperative abdominal leak, and not related to ofloxacin therapy. Intravenous/oral ofloxacin is effective and safe for the treatment of many difficult infections of the skin and skin structure, including those in diabetic subjects and in patients in whom previous parenteral therapy may have failed. PMID- 8419114 TI - Cyclosporine treatment for dermatomyositis/polymyositis. AB - We report one case of dermatomyositis and one case of polymyositis and scleroderma in which the patients' condition improved after treatment with cyclosporine (cyclosporine A). Both patients' conditions were unresponsive to systemic steroid therapy. The addition of cyclosporine to the regimen enabled us to taper the steroid dosage; the lesions resolved and the results of laboratory tests returned to normal. In nine of thirteen studies of dermatomyositis treated with cyclosporine reported in the literature, definite benefit was achieved and no serious toxicity was noted. PMID- 8419115 TI - Steroidogenesis-inducing protein stimulates protein-tyrosine kinase activity in rat Leydig cells. AB - Steroidogenesis-inducing protein (SIP) isolated from human ovarian follicular fluid stimulates steroid production in Leydig cells, human luteal cells, and rat adrenal cells. In addition, SIP is a potent mitogen that stimulates the proliferation of Leydig cells from immature rats to a greater extent than do LH/hCG and other known growth factors. We have shown previously that the actions of SIP on Leydig cells are independent of the adenyl cyclase-cAMP pathway. In the present study we have explored the possibility that SIP, like many growth factors, may exert its effects by activation of tyrosine kinase(s). Stimulation of Leydig cells isolated from immature rats with SIP resulted in an increase in the tyrosine phosphorylation of proteins that were detected with phosphotyrosine specific antibodies. The phosphorylation of a 90-kilodalton (kDa) protein band, a 65-kDa protein band, and a doublet at 140 kDa was apparent after 5 min. After 30 min, additional SIP-induced phosphotyrosine proteins were detected at 42, 44, 50, 80, 100, and 150 kDa. In addition to phosphorylation at tyrosine residues, all of the proteins isolated from SIP-stimulated cells were phosphorylated at threonine and serine residues. SIP-induced phosphoproteins recovered with phosphotyrosine specific antibodies were found to have associated protein-tyrosine kinase activity. The major substrate for this kinase activity in vitro was a 140-kDa protein, similar to one of the major phosphotyrosine-containing proteins induced by SIP treatment of intact cells. These observations suggest that SIP influences gonadal cell steroidogenesis and proliferation, presumably by activating cellular protein-tyrosine kinase(s) as part of a phosphorylation-based signalling pathway. PMID- 8419116 TI - Effect of vitamin D depletion on calcium transport by the luminal and basolateral membranes of the proximal and distal nephrons. AB - To study the effect of vitamin D on calcium (Ca2+) reabsorption by the kidney, we measured Ca2+ uptake by the basolateral and luminal membranes of proximal and distal tubules obtained from rabbits fed a vitamin D-deficient diet for 3 weeks. Results were compared to those obtained with a group of control animals fed a normal diet. Serum Ca2+ concentrations were comparable in both groups. In the control group, serum PTH, 25-hydroxyvitamin D3, and 1,25-dihydroxyvitamin D3 remained relatively stable. In the vitamin D-deficient animals, serum PTH levels slightly, but not significantly, increased, and the levels of vitamin D metabolites abruptly fell. Vitamin D depletion produced a 40% decrease in ATP dependent Ca2+ uptake by the basolateral membrane of the distal tubule. There was no change in the activity of the Na+/Ca2+ exchanger. A very significant effect was also observed in the luminal membrane of the distal tubule, where a 50% decrease in Ca2+ uptake was observed after the third week of vitamin depletion. Administration of 0.1 microgram 1,25-dihydroxyvitamin D3 16 and 2 h before death partially reestablished normal uptake. In contrast, no change in Ca2+ uptake could be detected in the basolateral or luminal membranes of the proximal tubule. These observations provide the first evidence of an effect of vitamin D on Ca2+ transport at both the basolateral and luminal membranes of the distal segment of the nephron. PMID- 8419117 TI - Effect of thyroid hormone on epidermal growth factor gene expression in mouse submandibular gland. AB - Epidermal growth factor (EGF) in mouse submandibular gland (SMG) is synthesized in the granular convoluted tubular (GCT) cells. The synthesis of EGF in SMG has been shown to be increased by thyroid hormone. This increase was attributed to the increase in EGF mRNA. Not known is how thyroid hormone increases the mRNA level. In the present study the effect of thyroid hormone administration on EGF gene expression in SMG was studied in hypothyroid mice. Hypothyroidism was induced by treating the mice with propylthiouracil. The amount of SMG EGF mRNA was markedly decreased in hypothyroid mice. Administration of T3 increased the mRNA in a dose-dependent manner. The increase in EGF mRNA by T3 was evident as early as 6 h after T3 administration. A nuclear run-off assay indicated that the induction of EGF gene expression by T3 is at a transcriptional level. Bromodeoxyuridine incorporation into GCT cells was not affected by T3 administration, suggesting that T3 does not cause the proliferation of these cells. In situ hybridization revealed that T3 increases EGF mRNA in GCT cells at a single cell level. These results suggest that thyroid hormone increases EGF gene transcription without affecting cellular proliferation. PMID- 8419118 TI - Suppression of GLUT4 expression in skeletal muscle of rats that are obese from high fat feeding but not from high carbohydrate feeding or genetic obesity. AB - In this study we determined whether alterations in the expression of GLUT4, the major insulin-regulatable glucose transporter, in skeletal muscle could explain the insulin-resistant glucose uptake characteristic of both dietary-induced and genetic obesity. GLUT1 expression was measured for comparison. To assess glucose transporter protein levels in dietary-induced obesity, postnuclear membranes were prepared from hindlimb muscle of Sprague-Dawley rats fed chow ad libitum (control), a high calorie/carbohydrate diet, or a high fat (80%) diet for 7 weeks. Immunoblotting revealed that GLUT4 protein levels decreased 34% in high fat-fed rats, but were unaltered in high calorie/carbohydrate-fed obese rats compared to control values. GLUT4 mRNA per DNA decreased 47% in muscle of high fat-fed rats compared to that in control or high calorie-fed rats; GLUT1 mRNA was reduced 31%, and actin mRNA tended to decrease (29%). To assess GLUT4 and GLUT1 expression in genetic obesity, similar studies were carried out in 5- and 20-week old lean and obese Zucker rats as well as in 20-week-old obese Zucker rats 36 h after streptozotocin injection to lower insulin levels. GLUT4 protein and mRNA levels were unaltered in hindlimb muscle of obese Zucker rats at either age or in the acutely diabetic state, whereas GLUT1 protein and mRNA levels decreased 40 45%. Comparison of these results with recent data in adipocytes demonstrates tissue-specific regulation of expression of GLUT4 and GLUT1. Thus, obesity due to high fat feeding, but not that due to high calorie/carbohydrate feeding or genetics, is associated with pretranslational suppression of GLUT4 expression in skeletal muscle. In at least some forms of obesity, the level of GLUT4 expression in muscle appears to be only one factor in, or may even be unrelated to, the degree of insulin-responsive glucose transport in vivo. PMID- 8419119 TI - Insulin-like growth factor-I regulation of renal 25-hydroxyvitamin D-1 hydroxylase activity. AB - Controversy exists regarding the role of GH and insulin-like growth factor-I (IGF I) in the modulation of calcitriol production. While their administration increases serum levels of 1,25-dihydroxyvitamin D, the mechanism remains unknown. Investigations have also implicated GH as a causal factor underlying renal 25 hydroxyvitamin D-1 alpha-hydroxylase activity [1(OH)ase] secondary to phosphate depletion. Thus, we investigated the effects of IGF-I on 1(OH)ase and the relationships between these actions and those of phosphate depletion. Our studies indicate that IGF-I administration to normal mice results in a dose-dependent (0 10 micrograms/h) increase in 1(OH)ase with maximum effects evident after 24 h, independent of changes in serum calcium, phosphorus, and glucose levels. Similarly, hormone administration to phosphate-depleted mice increases enzyme activity (6.06 +/- 0.96 vs. 13.97 +/- 1.67 fmol/mg.min) but to a level significantly greater than that achieved in normals (2.72 +/- 0.4 vs. 5.01 +/- 0.56 fmol/mg.min). Furthermore, the response represents an additive increment of the effects elicited by maximum doses of IGF-I and phosphate depletion, suggesting that the hormone- and phosphate-dependent enzyme stimulation occur by different mechanisms. Thus, our data establish that IGF-I stimulates renal 1(OH)ase activity in a time- and dose-dependent fashion. However, they do not support the hypothesis that IGF-I modulates the effects of phosphate-depletion on 1(OH)ase activity. Regardless, the documentation that IGF-I stimulates enzyme function provides an explanation for many observed physiological states associated with concomitant alterations of hormone levels and calcitriol production. PMID- 8419120 TI - Neuropeptide Y gene expression in the arcuate nucleus: sexual dimorphism and modulation by testosterone. AB - Neuropeptide Y (NPY) peptide concentrations in the arcuate nucleus have recently been shown to be modulated by gonadal steroids in the male rat. The present study was designed to determine whether NPY messenger RNA (mRNA)-synthesizing cells in the arcuate nucleus (Arc) of the male rat are regulated by testosterone (T) and whether there is a sexual dimorphism in the expression of the NPY gene in this region. In situ hybridization and quantitative autoradiography were used to assess the level of NPY gene expression in the Arc. In the first experiment, NPY mRNA levels were measured in the Arc of intact, castrated, and castrated male rats treated with T to maintain physiological (1.3 +/- 0.1 ng/ml) and supraphysiological (5.3 +/- 0.4 ng/ml) plasma levels of T. A 2-week castration produced a modest but significant decrease in NPY mRNA levels in the Arc (P < 0.05). Replacement with either physiological or supraphysiological levels of T prevented the effect of castration on NPY gene expression, and there was no further potentiation of NPY gene expression in those animals that received high levels of T. In the second experiment, NPY gene expression was compared throughout the Arc between intact male and female rats at 1800 h on the afternoon of proestrus. Comparison of NPY gene expression throughout the rostro-caudal extent of the Arc showed that male rats had significantly more NPY mRNA containing cells than female rats (P < 0.01). This difference was most strikingly observed in the caudal portions of the nucleus (3.80 mm caudal to bregma). No difference was detected in the mean levels of NPY gene expression in the Arc between male and female rats. These data demonstrate that 1) NPY gene expression throughout the arcuate nucleus is modulated by T in male rats, and 2) a marked regional sex difference exists in the distribution of NPY mRNA-containing cells in the caudal extremity of the Arc. It is hypothesized that gonadal hormones may exert both organizational and activational effects upon NPY neurons in the Arc. PMID- 8419121 TI - Immunocytochemical analysis of androgen receptor along the ducts of the separate rat prostate lobes after androgen withdrawal and replacement. AB - Autoregulation of androgen receptors (AR) in the rat prostate gland has previously been shown to be lobe specific. Saturation ligand-binding assays revealed that AR fell to very low levels in the ventral and dorsal prostate within 7 days after castration, whereas lateral lobe AR were present at intact levels at that time. To study this differential response further, we herein analyzed AR in the separate prostate lobes by indirect immunocytochemistry after castration and testosterone replacement to adult rats. The ventral and dorsal lobes each contain one type of duct, whereas the lateral lobe is composed of two ductal systems, which were separated as LP1 and LP2. Frozen ducts were sectioned longitudinally to reveal the proximal-distal orientation. Sections were stained for AR with PG-21 antibody against rat AR. Within 2 days after castration, ventral and dorsal lobe immunoreactive nuclear AR was markedly decreased in staining intensity in the secretory epithelium compared to that in the intact rat and was absent in all stromal cells. Epithelial immunostaining continued to decline to a weak punctate nuclear signal by day 7, which further dissipated by day 21. Proximal and intermediate regions of the ducts were largely devoid of AR signal after castration, whereas residual nuclear staining was most apparent in epithelial cells of the distal ductal region. By day 7 and beyond, specific cytoplasmic staining for AR was also observed in distal tip epithelial cells. In the lateral lobe, LP1 ducts rapidly lost all AR immunostaining upon androgen withdrawal. In marked contrast, epithelial cells in the LP2 ducts retained AR immunostaining at all time points after androgen withdrawal at a signal intensity equivalent to that in the intact animal. Within 15 min after testosterone injection to 14-day castrate rats, considerable nuclear AR immunostaining was apparent within the distal tip epithelial cells of the ventral, dorsal, and lateral LP1 lobes. Cytoplasmic signal was noticeably reduced at this time. With increasing time after continued testosterone replacement, nuclear AR signal intensity increased, so that by 72 h, nuclear AR signal in all secretory epithelial cells approached the staining intensity observed in intact rats. AR immunostaining returned to smooth muscle and fibroblastic stromal cells within 1 3 days after testosterone replacement. In summary, immunodetectable AR declined in the ventral, dorsal, and LP1 prostate ducts after castration-induced androgen withdrawal and returned upon testosterone replacement, which further indicates that androgen up-regulates AR protein within these prostatic regions.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419122 TI - The effect of testicular macrophages and interleukin-1 on testosterone production by purified adult rat Leydig cells cultured under in vitro maintenance conditions. AB - A functional interaction between testicular macrophages and Leydig cells has been suggested. The present study attempts to clarify the interaction between purified Leydig cells and macrophages from adult male rats in coculture, employing culture conditions that maintain Leydig cell steroidogenic responsiveness in vitro. Basal Leydig cell testosterone production over 24 h was not significantly affected by coculture with macrophages, but an inhibitory effect of testicular macrophages on testosterone production by Leydig cells over 24 h was observed in the presence of increasing doses of LH from 0.125 ng/ml up to a maximally stimulating dose of 8 ng/ml. A consistent inhibitory effect was observed over a range of Leydig cell testicular macrophage coculture ratios from 0.5:1 to 4:1 in the presence of LH (8 ng/ml). A similar inhibitory effect on maximal LH-stimulated Leydig cell testosterone production over 24 h was observed when Leydig cells were cocultured with peritoneal macrophages. Conditioned medium collected from testicular or peritoneal macrophage cultured for 24 h also inhibited LH-stimulated Leydig cell testosterone production, indicating that the effect of the macrophages was mediated by a secreted product. Inhibition of LH-stimulated testosterone production was observed also when Leydig cells were cultured in the presence of testicular macrophages for 24 h before maximal LH stimulation (8 ng/ml) for a further 24 h. Human recombinant interleukin-1 alpha and interleukin-1 beta (0.5 10 U/ml) did not significantly alter basal or LH-stimulated Leydig cell testosterone production at 24, 48, or 72 h of culture. The specific binding of 125I-human CG to Leydig cells was not affected by testicular macrophage conditioned medium. These data demonstrate that testicular and peritoneal macrophages inhibit LH-stimulated Leydig cell testosterone production in coculture through secreted factors, acting distal to the LH receptor, and provide further support for paracrine interactions between these cell types. PMID- 8419123 TI - Tissue-specific regulation of shell gland calbindin D28K biosynthesis by estradiol in precociously matured, vitamin D-depleted chicks. AB - Provision of Ca2+ for egg shell calcification in the avian uterus [egg shell gland (ESG)] derives mostly from vitamin D-dependent intestinal Ca2+ absorption from the diet. Ca2+ absorption is strongly linked to the intestinal vitamin D dependent calbindin D28K (D28K) concentration. The laying hen ESG also contains D28K, and again, Ca2+ transport into the shell appeared to be linked to the ESG D28K concentration. However, evidence is now presented that ESG D28K synthesis may be estradiol (E2) dependent and vitamin D independent under certain conditions. One-day-old female chicks fed a vitamin D-free diet for as long as 6 weeks and then repeatedly injected im with E2 for up to 3 more weeks developed frank rickets, but possessed precociously matured reproductive tracts. While the tiny presumptive ESGs of nonestrogenized vitamin D-depleted chicks were devoid of D28K, the highly developed ESG, including the isthmus, of estrogenized chicks contained D28K. The ESGs of nonestrogenized, vitamin D-replete chicks also exhibited no development or detectable D28K. Regardless of whether vitamin D depleted or replete, estrogenized chick ESG contained similar D28K and D28K mRNA concentrations. Immunohistochemical techniques showed that the endometrial cellular localization of both D28K and Ca(2+)-ATPase (Ca2+ pump) in estrogenized chicks was similar to that in mature laying hens. There was no trace of D28K, nor was there any stimulation of Ca2+ absorption, in duodenum of vitamin D-free, immature chicks regardless of E2 treatment. As expected, both D28K and D28K mRNA were present in vitamin D-replete chick duodenum. We conclude that in E2-treated chicks, ESG D28K gene expression may be vitamin D independent and E2 dependent. This is the first clear demonstration of hormone-dependent tissue-specific D28K gene expression in the chick. PMID- 8419124 TI - Secretory patterns and rates of gonadotropin-releasing hormone, follicle stimulating hormone, and luteinizing hormone revealed by intensive sampling of pituitary venous blood in the luteal phase mare. AB - We used our unique nonsurgical technique for collecting pituitary venous (pit) blood to study GnRH, FSH, and LH secretion patterns in midluteal phase mares. This method does not perturb endocrine function and allows continuous monitoring of GnRH and gonadotropin (Gn) secretion, determination of the amount of GnRH perfusing gonadotropes, and direct measurements of the amounts of Gn secreted. In a total of 80 h of 5-min sampling in four mares, eight Gn peaks occurred; however, more frequent sampling was needed to define secretory events precisely. Therefore, pit blood was collected continuously and split into 30-sec segments in six mares. To ensure a peak during sampling, the opioid antagonist naloxone was given after 4-6 h of sampling to try to replicate a physiological signal for GnRH release. Naloxone induced Gn peaks in jugular blood that were indistinguishable in amplitude from spontaneous peaks. Intensive sampling of pit blood showed that jugular peaks reflected major episodes of GnRH and Gn secretion lasting 30-55 min, which were similar in profile whether naloxone induced or spontaneous and consisted of a train of three to six peaks of diminishing amplitude. Peaks of GnRH and, less often, Gn also occurred outside major episodes. Despite markedly variable size, GnRH peak maxima were correlated with the amount of LH and FSH secreted in concurrent peaks. Likewise, cross-correlation analyses (n = 960 samples/mare) showed close correspondence between patterns of GnRH and secreted FSH and LH. The delay (+/- SEM) between GnRH and Gn maxima was 0.62 +/- 0.18 min for LH and 0.18 +/- 0.22 min for FSH. The majority of GnRH and Gn peaks were concurrent; however, 34.7% of GnRH peaks occurred without Gn peaks. These peaks had a lower amplitude than those with Gn peaks (P < 0.001). For Gn, secretion (i.e. ratio between pit and jugular concentrations, > 1.5) continued at a low level for 40 +/- 9% (LH) or 64 +/- 14% (FSH) of the time between Cluster-defined peaks during the basal period. We conclude that in the luteal phase 1) the predominant mode of GnRH and Gn secretion is as concurrent, large amplitude, prolonged episodes that appeared to be the summation of a train of peaks; and 2) a GnRH dose-Gn response relationship operates endogenously. This along with the synchronicity of secretion patterns of the three hormones suggest that GnRH is the major secretagogue for both LH and FSH. PMID- 8419125 TI - Loss of lutropin/human choriogonadotropin receptor messenger ribonucleic acid during ligand-induced down-regulation occurs post transcriptionally. AB - Previous studies have demonstrated that ligand-induced down-regulation of the LH/hCG receptor in rat corpus luteum results in a loss of ligand-binding activity and a parallel decline in steady state levels of receptor mRNA. The purpose of the present study was to determine whether this loss of receptor mRNA during receptor down-regulation is due to inhibition of transcription or increased degradation of the receptor mRNA. To differentiate between these two possibilities, nuclear run-off assays were performed to study transcription rates of nuclei isolated from control and hCG down-regulated rat ovaries. A 750-mer LH/hCG receptor cDNA probe, spanning the carboxy-terminal region of the peptide and the 3'-untranslated region of the receptor cDNA, was constructed by polymerase chain reaction. RNA transcripts were synthesized from existing nuclear RNA from control and down-regulated nuclei and hybridized to the receptor cDNA probe immobilized on nylon membranes and subjected to autoradiography. Hybridization to actin cDNA was also run alongside as a control. The results of run-off transcription analysis indicated that during down-regulation, the transcription rates of LH/hCG receptor mRNA are not decreased compared to those in saline-treated controls. Although no decrease in the transcription rates of the receptor mRNA was seen in the down-regulated state, the steady state levels of the receptor mRNA showed a decline when assayed by either solution hybridization or Northern blot analysis. Furthermore, hCG induced down-regulation appears to increase the transcriptional activity of the nuclei. Estimates of steady state LH/hCG receptor mRNA turnover rates indicate that the half-life of the message in down-regulated ovaries is markedly reduced compared to that in the control. It is concluded that the loss of steady state LH/hCG receptor mRNA is not due to a decrease in transcription, but probably represents an increased degradation of the receptor mRNA. PMID- 8419126 TI - Apoptosis during luteal regression in cattle. AB - The present studies were conducted to evaluate whether apoptosis occurs during spontaneous and prostaglandin F2 alpha (PGF2 alpha)-induced luteolysis and, if so, to determine the relationship between the onset of luteolysis and oligonucleosome formation (a characteristic of apoptosis). In the first study, nine normally cycling heifers were ovariectomized (ovx) during the midluteal phase (day 10 or 15; day 0 = estrus) or after luteal regression (day 19; n = 3/time point). While there was no evidence of oligonucleosome formation in DNA from corpora lutea (CL) collected on days 10 and 15, each CL collected on day 19 exhibited DNA fragmentation, represented by distinct bands of DNA in approximately 185-basepair multiples. In the second study, heifers were ovx (n = 5; controls) or given 25 mg PGF2 alpha 15-16 days after estrus. Heifers receiving PGF2 alpha were subsequently ovx 4, 8, 12, 24, or 48 h (n = 5/time point) after the injection of PGF2 alpha. The concentration of progesterone in venous sera collected at ovx was not different (P > 0.20) in control and 4 h groups, but was decreased (P < 0.01) in the 8, 12, 24, and 48 h groups. Total CL weight (mean +/- SEM; grams) did not change (P > 0.10) from 0 h (controls) to 24 h after injection (range, 3.2 +/- 0.5 to 4.1 +/- 0.6), but decreased (P < 0.06) to 2.0 +/- 0.3 at 48 h. With ethidium bromide (EtBr) staining, no oligonucleosome formation was detected in CL collected from 0-12 h after PGF2 alpha injection. However, pronounced oligonucleosome formation was observed in all 10 CL collected 24 and 48 h after the injection of PGF2 alpha. The absence of oligonucleosomes in 0 and 4 h samples was confirmed by the more sensitive technique of 3'-end labeling of DNA fragments. Some samples in both the 8 and 12 h groups had slight oligonucleosome formation, while all samples in the 24 and 48 h groups showed evidence of intense oligonucleosome formation. Histological analysis of tissue sections indicated an increase (P < 0.001) in the percentage of degenerated luteal cells in the 24 and 48 h groups compared to that in the 0-12 h groups. These data indicate that apoptosis occurs during both spontaneous and PGF2 alpha induced luteal regression in cattle; however, apoptosis, as indicated by oligonucleosome formation, is not apparent until after serum progesterone concentrations have begun to decrease.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419127 TI - Opposing influences of dexamethasone and retinoic acid on adenylate cyclase activity in ROS 17/2.8 cells. AB - Exposure of ROS 17/2.8 cells to dexamethasone (DEX) or retinoic acid (RA) increases and decreases, respectively, adenylate cyclase activity (ACA) in response to isoproterenol, forskolin, guanylylimidodiphosphate, or NaFl. Despite dramatic changes in ACA, there were no significant changes in levels of cholera toxin- or pertussis toxin (PT)-dependent ADP-ribosylation of membranes prepared from cells after DEX or RA exposure as compared to controls. Similarly, immunochemical detection of alpha S, alpha i1-3, and alpha O, as well as Northern blot analysis of messenger RNA for each of the respective GTP binding proteins, also failed to demonstrate an influence of DEX or RA when contrasted with controls. In a novel use of the cyc- reconstitution assay, wherein the influence of inhibitory guanine nucleotide binding proteins in the extracts of control, DEX , and RA-treated membranes is removed by a previous 24-h incubation with PT in the intact cell, we demonstrate that this PT treatment markedly enhances ACA in the cyc- reconstitution assay for all three preparations, but that the fold increase due to PT-treatment is greatest in RA-treated cells. The greater magnitude of the effect of PT on RA-treated ROS 17/2.8 cells, in the absence of any obvious quantitative changes in the levels of the PT substrates, suggests that the effect of RA on ROS 17/2.8 cells appears to be an augmentation of the influence of inhibitory guanine nucleotide binding proteins, ultimately leading to reduced ACA. PMID- 8419128 TI - Estimation of the protein content of thyroid hormone receptor alpha 1 and beta 1 in rat tissues by western blotting. AB - Recent studies of the expression of c-erbA/thyroid hormone receptor (TR) mRNAs have revealed a dissociation between T3-binding activity and the behavior of the mRNAs that code the functional TRs in some tissues. Compared with T3-binding activity, TR(alpha 1 + beta 1) mRNA is disproportionally high in the brain and low in the liver. Using anti-TR antiserum, 4BII, which recognizes TR alpha 1 and beta 1, but not the alpha 2-variant, we measured TR protein content in rat tissues by Western blotting. Two protein bands of 47 and 55 kilodaltons (kDa) were specifically identified as TR proteins. The positions of the in vitro transcription/translation products of c-erbA/TR alpha 1 and beta 1 cDNA on the gel were consistent with those of the 47- and 55-kDa bands, respectively. The 47- and 55-kDa proteins in nuclear proteins extracted with 0.4 M KCl from rat tissues were analyzed by Western blotting, and the intensity of TR protein bands in each tissue was measured by a densitometer. The relative TR protein concentration was highest in liver, followed by brain, kidney, and testis. We compared the TR protein level measured by Western blotting with the maximal T3-binding capacity (Cmax) in the same aliquot of samples from liver and brain. Both the TR protein level and the Cmax in the brain were about 40% of those in the liver, suggesting that the Cmax per receptor molecule is constant in these two tissues, and an abundant amount of functional TR proteins exists in the liver, corresponding to the high level of T3-binding activity. PMID- 8419129 TI - Uterine epithelial cell secretion of interleukin-1 alpha induces prostaglandin E2 (PGE2) and PGF2 alpha secretion by uterine stromal cells in vitro. AB - Uterine epithelial cells (UEC) were isolated from cycling mice and cultured on Matrigel-coated nitrocellulose filters to determine their ability to secrete interleukin-1 alpha (IL-1 alpha) in response to ovarian steroids and induce prostaglandin (PG) secretion by uterine stromal cells (USC). UEC cultured in a polarized manner secreted IL-1 alpha with an 8- to 10-fold apical vs. basal preference, as determined by an enzyme-linked immunosorbent assay. There was no effect of 17 beta-estradiol, progesterone, or 17 beta-estradiol plus progesterone on IL-1 alpha secretion by UEC. The mean total IL-1 alpha secreted to the apical and basal secretory compartments over the 24-h incubation period was 0.8 +/- 0.16 and 0.07 +/- 0.05 ng/2 x 10(5) cells, respectively. Cytokine bioactivity, as determined by [3H]thymidine incorporation into D10 cells in response to UEC conditioned medium, paralleled the pattern of IL-1 alpha secretion observed using the immunoassay. In addition to the in vitro secretion of IL-1 alpha by polarized UEC, pooled uterine fluid collected from proestrous stage mice contained IL-1 alpha at a concentration of 0.7 ng/ml, indicating that IL-1 alpha is released into the uterine lumen in vivo. Coculture with UEC or treatment with conditioned medium from either the apical or basal UEC secretory compartments induced a several-fold increase in the secretion of PGE2 and PGF2 alpha by USC. Relative to untreated USC, PGE2 was induced to a greater extent than PGF2 alpha. The addition of polyclonal anti-IL-1 alpha significantly inhibited the ability of UEC conditioned medium and UEC coculture to induce PG secretion by USC. In addition, mouse recombinant IL-1 alpha added at a concentration similar to that secreted by UEC stimulated USC PGE2 and PGF2 alpha secretion in a manner similar to that observed with UEC coculture. Experiments designed to determine the cell type specificity of the induction of PG secretion by USC indicated that conditioned medium from a human UEC line (RL95), a rat prostate epithelial cell line (E4), and a mouse fibroblast cell line (10T1/2) induced PG secretion to an extent that paralleled their ability to induce D10 cell proliferation. The present results demonstrate the ability of UEC to secrete IL-1 alpha in a vectorial manner. Soluble products secreted by UEC are capable of stimulating PGE2 and PGF2 alpha secretion by USC, and IL-1 alpha appears to be a significant factor contributing to this effect.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419130 TI - Inhibin subunit gene expression and distribution in the ovaries of immature, young adult, middle-aged, and old female rats. AB - In young adult female rats, the patterns of inhibin subunit mRNA expression during the estrous cycle are regulated by cyclic changes in gonadotropin secretion and follicular development. Since there are distinct alterations in profiles of both hormone secretion and folliculogenesis during the reproductive lifespan of the female rat, we have characterized the gene expression and distribution of inhibin subunit mRNAs in immature (22 days), young adult cyclic (3-4 months), middle-aged cyclic (9-10 months), and old (12-13 months) persistent estrous (PE) rat ovaries. Northern blot analyses revealed that in contrast to young adult cyclic rats, inhibin alpha- and beta A-subunit mRNA levels in the ovaries of middle-aged cyclic rats remained substantially elevated after the proestrous gonadotropin surges and ovulation. Likewise, acyclic immature and old PE rats showed high levels of inhibin alpha- and beta A-subunit transcripts in their ovaries. In situ hybridization analyses demonstrated that inhibin alpha- and beta A mRNAs were abundantly expressed in maturing follicles of both young and middle-aged cyclic females, while high levels of alpha-subunit transcripts were only detected in the ovarian stroma of middle-aged animals. The ovaries of old PE rats had numerous large cystic follicles (with variable layers of granulosa cells) and few degenerating cyst-like structures (completely devoid of granulosa cells). Inhibin subunit transcripts were expressed abundantly in both the granulosa (alpha and beta A-subunits) and theca interna (alpha-subunit only) layers of large follicles, but were absent from degenerating cysts devoid of granulosa cells. The ovarian stroma of PE rats also expressed very high levels of inhibin-alpha, but not beta A mRNA. The ovaries of immature rats contained large numbers of uniformly developing secondary follicles. High levels of inhibin-alpha mRNA were expressed homogeneously in the granulosa layer of all growing follicles, whereas inhibin beta A mRNAs were only detected in selectively larger follicles with multiple layers of granulosa cells. Hormone RIAs of serum samples from these same groups of animals showed that basal levels of serum FSH were substantially higher in immature, middle-aged cyclic, and old PE rats than in young adult rats. These results demonstrate that enhanced ovarian inhibin subunit gene expression in the female rat is associated with increased serum FSH levels regardless of chronological age. On the other hand, aging appears to selectively enhance inhibin alpha, but not beta A, gene expression in the ovarian stroma, such that it may gradually become a major secondary site of alpha-subunit mRNA production in addition to the follicular compartments. PMID- 8419131 TI - Cellular events involved in hormonal control of receptor-mediated endocytosis: regulation occurs at multiple sites in the low density lipoprotein pathway, including steps beyond the receptor. AB - We used biochemical and quantitative structural approaches to analyze hormonal regulation of receptor-mediated endocytosis of human low density lipoprotein (LDL) by differentiating rat granulosa cells in culture. We studied uptake and metabolism of 125I-labeled LDL and distribution of gold-labeled LDL in nontreated and FSH- or FSH/androstenedione-treated granulosa cells. FSH and androstenedione worked together to enlarge the segment of the granulosa cell population capable of accumulating LDL and to increase the number of LDL receptors at the cell surface. More importantly, FSH affected postreceptor aspects of the LDL pathway, including 1) accelerating the apparent rate of LDL internalization, and 2) decreasing the time needed for LDL to reach lysosomes. In both cases we eliminated the possibility that the observed effects merely reflected differences in receptor numbers between hormone-treated and nontreated cells. FSH also increased the number of both gold-labeled lysosomes per cellular profile and gold particles per lysosome. In all, FSH expanded the capacity of the LDL pathway to accommodate more intracellular ligand by augmenting the numbers of organelles participating in the endocytic process. However, it did not affect cell size, sizes of individual organelles comprising the LDL pathway, or numbers of gold particles per organelle (except lysosomes). Our findings indicate that hormonal regulation of the LDL pathway extends beyond simply causing expression of cell surface LDL receptors to encompass postreceptor events, including enhancing the apparent rates at which ligand is internalized and transported to lysosomes. PMID- 8419132 TI - Jejunal/kidney glucose transporter isoform (Glut-5) is expressed in the human blood-brain barrier. AB - The recently cloned Glut-5, glucose transporter isoform, is expressed in human jejunum and kidney. Employing previously characterized polyclonal antibodies directed towards the C-terminus region of the derived human Glut-5 peptide and Western blot analysis, a 50-55 kilodalton Glut-5 protein was detected in adult human brain homogenates. The amount of Glut-5 protein in brain was 4-fold lower when compared to the levels in adult kidney. Immunohistochemical analysis using cerebral and cerebellar sections demonstrated Glut-5 immunoreactivity in only some of the Glut-1 and factor VIII-positive brain microvascular endothelial cells, the intravascular red and white blood cells being negative. This selective localization pattern was confirmed by the 5-fold enrichment of Glut-5 vs. a 20 fold enrichment of Glut-1 in an isolated human cerebral cortical microvascular preparation, when compared to whole cerebral homogenates. We conclude that Glut-5 is localized in the endothelial cells of human brain microvasculature. Unlike other fructose using tissues, where Glut-5 may subserve the role of a fructose carrier, in brain where fructose is not used as a substrate, Glut-5 may transport glucose alone. This role of Glut-5 in conjunction with the previously characterized brain endothelial Glut-1 and Glut-3 needs further elucidation. PMID- 8419133 TI - Isoelectric charge of recombinant human follicle-stimulating hormone isoforms determines receptor affinity and in vitro bioactivity. AB - Recombinant human FSH (rhFSH) was obtained by expressing the human FSH alpha- and beta-subunit complementary DNAs in the chinese hamster ovary cell line. Isoforms of rhFSH were resolved into specific isoelectric (pI) fractions by chromatofocusing. rhFSH isoforms ranged from pI 3.0-5.5 with a modal value of pI 4.2. Analysis of the biological activity of specific pI isoforms of rhFSH was undertaken using both the rat granulosa cell aromatase (in vitro) bioassay and a RRA. More acidic isoforms (e.g. pI 3.5) showed significantly lower affinity (P < 0.05) for rat testicular FSH receptors than did the less acidic isoforms (e.g. pI 4.8). Consistent with the receptor binding affinity data, the more acidic fractions resulted in significantly less activation (P < 0.05) of rat granulosa cell aromatase activity, as measured by estrogen production, than did the less acidic isoforms. The observed bioactivities and their correlation with the pI values of the rhFSH isoforms are consistent with observations of differing bioactivities seen in both pituitary and urinary FSH isoforms. These results demonstrate that rhFSH, made in the chinese hamster ovary cell line, is both biologically active and has isoform profiles, and presumably carbohydrate structures, that closely resemble those seen in natural hFSH. PMID- 8419134 TI - Impairment of the selenoenzyme type I iodothyronine deiodinase in C3H/He mice. AB - Type I iodothyronine deiodinase (ID-I) activity is impaired in C3H/He (C3H) mice compared with BALB/c and C57BL/6N (C57) mice. In this study we compared ID-I activity and protein labeling with N-bromoacetyl(-)[125I]T3 (BrAc[125I]T3) or 75Se in liver microsomes of C3H and C57 mice. Hepatic ID-I activity in C3H mice was highly variable with a median of only 18% of that in C57 mice. However, C3H mice had normal serum T4 and T3 levels, although serum reverse T3 was increased. The 28-kilodalton (kDa) ID-I protein was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis of BrAc[125I]T3-labeled microsomes. Labeling of this protein was virtually undetectable in C3H samples with low enzyme activity. ID-I activity in liver microsomes was strongly decreased in Se deficient mice, which was paralleled by a drastic decrease in BrAc[125I]T3 labeling of the 28-kDa band compared with control mice. Labeling of ID-I with 75Se was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of liver microsomes of [75Se]selenite-injected mice. 75Se labeling of the 28-kDa band was markedly higher in Se-deficient than in control mice and was also markedly higher in C57 than in C3H mice. Finally, liver ID-I messenger RNA (mRNA) was measured on Northern blots using a rat ID-I complementary DNA probe. Messenger RNA levels correlated strongly with ID-I activity, showing a significant decrease in C3H mice. We conclude that in mice, like in rats and humans, ID-I is a selenoprotein. ID-I activity is impaired in C3H mice because of decreased transcription of the ID-I gene or reduced stability of the mRNA. PMID- 8419135 TI - Failure of bilateral paraventricular nuclear lesions to cause hypothalamic hypothyroidism in fetal sheep. AB - Concentrations of T4 indicative of a hyperthyroid state are present in fetal sheep plasma between 100 days gestational age (dGA) and parturition. Fetal pituitary stalk section studies indicate that, as in adults, these high fetal plasma T4 concentrations during pregnancy are controlled by the hypothalamus. We compared peripheral plasma T4 concentrations in fetal sheep with bilateral hypothalamic paraventricular nuclear (PVN) lesions (lesion group; n = 5) to fetal sheep with sham-PVN lesions (sham group; n = 4) at 131 and 146 dGA in the lesion group or at term (mean +/- SEM, 146 +/- 0.9 dGA) in the sham group. Bilateral hypothalamic PVN lesions or sham lesions were placed at 118-122 dGA. Baseline blood samples were taken between 1100-1500 h at 131 dGA in both groups, at term in the sham group, and at 146 dGA in the lesion group. In control sheep, TRH cells were found in the PVN and in a number of extra-PVN sites, and the median eminence received abundant TRH axons. In the lesion group, complete destruction of the PVN bilaterally was confirmed by histology. Extra-PVN TRH neurons remained intact in the lesioned sheep, and axons to the median eminence were reduced, but not eliminated. T4 concentrations in fetal plasma were not different in the lesion group and the sham group at 131 dGA (81 +/- 7 vs. 92 +/- 19 ng/ml) or at term (112 +/- 35 vs. 79 +/- 15 ng/ml), respectively. In contrast, fetal plasma concentrations of cortisol, which were not different in lesion and sham group fetuses at 131 dGA (1.7 +/- 0.3 vs. 1.0 +/- 0.2 ng/ml, respectively), were greatly reduced (P < 0.05) at 146 dGA in the lesion group compared to those in the sham group at term (2.0 +/- 0.5 vs. 58.8 +/- 11.5 ng/ml). We conclude that unlike in adult rats, the ovine fetal PVN is not required to maintain normal plasma T4 concentrations. The many TRH-positive cells that lie outside of the PVN in the fetal sheep appear to enable PVN-lesioned fetuses to remain euthyroid fetuses. PMID- 8419136 TI - Localization of estrogen receptor messenger ribonucleic acid in rhesus monkey uterus by nonradioactive in situ hybridization with digoxigenin-labeled oligodeoxynucleotides. AB - Previous immunocytochemical studies indicate that receptor regulation varies in different uterine cell types. In primates, progesterone (P) suppresses estrogen receptor (ER) in glandular epithelial cells in the functionalis, but fails to suppress ER in the glandular epithelial (GE) cells of the basalis. P also fails to suppress ER in the perivascular stromal and smooth muscle cells of the spiral arteries in the functionalis. We used nonradioactive in situ hybridization to determine whether similar cell type differences occur at the ER mRNA level. We used digoxigenin-labeled oligodeoxynucleotides (oligo-DNAs; 45-mer) as probes and detected the hybrids immunocytochemically with horseradish peroxidase-labeled antidigoxigenin antibody. This technique can discriminate between positive and negative cells in closely packed histological associations. In spayed monkeys, most of the GE cells as well as endometrial stromal cells were positive for ER mRNA, while all vascular smooth muscle, endothelium, and perivascular stromal cells were negative. Estradiol treatment for 14 days markedly increased ER mRNA staining in the GE cells, most stromal cells, and the vascular smooth muscle and perivascular stromal cells of spiral arteries in the functionalis. However, in the basalis, these components of the spiral arteries were negative as were the small basal arteries of the basalis. In most positive cells, ER mRNA was not homogeneously distributed in the cytoplasm, but, rather, was concentrated in their perinuclear regions. The GE cells in the basalis had especially intense concentrations of perinuclear signal at their apical poles. After sequential estradiol plus P treatment, the signal was greatly reduced in the GE cells of the functionalis, but not in the GE cells of the basalis or in the vascular smooth muscle or perivascular stromal cells of the spiral arteries of the functionalis. In myometrium, ER mRNA was localized to the perinuclear region of smooth muscle cells, but the staining intensity was not dramatically affected by hormonal manipulation. Unexpectedly, we observed clusters of stromal cells characterized by extremely high positive signals for ER mRNA ("hot cells") at the endometrial/myometrial border and deeper in the connective tissue of the myometrium, although such cells did not express high levels of ER protein. In general, however, the cellular distribution of ER mRNA and its hormonal regulation paralleled those of ER protein. PMID- 8419137 TI - Ovulatory effect of interleukin-1 beta on the perfused rat ovary. AB - The involvement of cytokines derived from ovarian and hematopoietic cells have been suggested in the cyclic events of the ovary. In the present study the effects of two cytokines, interleukin-1 beta (IL-1 beta) and IL-2, on ovulation, steroidogenesis, and prostaglandin (PG) production were explored in rat ovaries perfused in vitro. Ovaries of equine CG (20 IU)-primed immature rats were perfused in a recirculating system for 20 h, and samples were taken for analysis of progesterone, estradiol, prostaglandin E (PGE), and PGF2 alpha. The number of ovulations was estimated by counting the number of oocytes released into the ovarian bursae. Unstimulated ovaries did not ovulate, whereas the addition of LH (100 ng/ml) resulted in 3.4 +/- 0.6 ovulations/treated ovary. The addition of human recombinant IL-1 beta (4 ng/ml) induced ovulation (1.6 +/- 0.4) and increased the LH-induced ovulation rate 3-fold (9.8 +/- 0.5). The addition of human recombinant IL-2 (40 ng/ml) did not induce ovulation and did not affect the LH-induced ovulation rate. Ovarian release of progesterone and PGF2 alpha was increased by IL-1 beta, but estradiol and PGE release was not affected. IL-2 did not affect steroidogenesis or PG release. To elucidate whether the IL-1 beta stimulated ovarian synthesis of PGF2 alpha was crucial for the ovulatory effect of IL-1 beta, experiments were performed in the presence of indomethacin (5 micrograms/ml), an inhibitor of PG synthesis. Indomethacin (5 micrograms/ml) inhibited LH-induced ovulation almost completely (one ovulation in one of the five treated ovaries), but did not affect the IL-1 beta-induced ovulation rate (1.4 +/- 0.2). The number of ovulations in the group treated with LH and IL-1 beta was significantly reduced (3.2 +/- 0.6) in the presence of indomethacin. These data demonstrate that IL-1 beta induces ovulation in the rat ovary, and this effect may be partly mediated by the increased production of the ovulatory mediator progesterone. PMID- 8419138 TI - Inhibin and activin as paracrine/autocrine factors. PMID- 8419139 TI - Unsustained dipsogenic response to chronic central infusion of angiotensin-III in spontaneously hypertensive rats. AB - We evaluated the chronic effect of angiotensin-III (AIII) in the promotion of drinking behavior in spontaneously hypertensive (SH) and normotensive Wistar Kyoto (WKY) rats, using conscious, freely moving, male, adult animals that had been instrumented with an intracerebroventricular (icv) cannula connected to an osmotic mini-pump for 7-day infusion. Chronic icv infusion of AIII (5 or 10 pmol/min) elicited robust, dose-dependent, and Ile7-AIII (100 pmol/min; as specific antagonist)-reversible dipsogenesis in both SH and WKY rats, with higher water intake in the former strain. However, the drinking response in the SHRs exhibited a sharp drop after 3 days of AIII infusion, during which acute AIII (80 pmol, icv) challenges also failed to induce dipsogenesis. Chronic icv infusion of bestatin (150 pmol/min), an aminopeptidase-B inhibitor, did not by itself discernibly affect basal drinking. When combined with AIII (5 or 10 pmol/min), however, bestatin, respectively, suppressed and augmented the dipsogenic response of SH and WKY rats to the heptapeptide. These results suggest that chronic administration of AIII did not produce sustained drinking behavior in SHRs, possibly because of the development of early desensitization of the angiotensin receptors. PMID- 8419140 TI - Identification of biologically active epidermal growth factor precursor in human fluids and secretions. AB - A biologically active form of the epidermal growth factor (EGF) precursor has been detected in human fluids and secretions. The secreted protein identified in human urine and milk has an apparent molecular mass of 160-170 kilodaltons and exhibits an affinity for the glycosaminoglycan heparin. More importantly, the secreted EGF precursor is capable of activating the intrinsic tyrosyl kinase activity of the EGF receptor. Our results demonstrate that the soluble form of the precursor is generated from the membrane-anchored form by a processing step that takes place at the cell surface and involves truncation of the cytoplasmic and transmembrane domains of the intact EGF precursor. The findings support the hypothesis that the secreted 160- to 170-kilodalton EGF glycoprotein that accumulates in urine and milk is proteolytically derived from the plasma membrane spanning precursor expressed in the kidney and mammary gland. PMID- 8419141 TI - The presence of functional estrogen receptors in intestinal epithelial cells. AB - Reproductive and maturational nutritive needs are examples of situations in which alterations in circulating concentrations of estrogens are associated with changes in intestinal epithelial function. However, it is not clear that any of these effects is due to direct interaction of estrogen with intestinal epithelial estrogen receptors (ER). The experiments reported here were designed to determine whether the small intestinal epithelium contains functional ER and might, therefore, be an estrogen-responsive tissue. IEC-6 cells, a non-transformed line of cells isolated from rat small intestinal crypts, were used for many of the experiments, because they provide a pure preparation of crypt epithelial cells. IEC-6 cells were found to exhibit specific saturable binding of estradiol with a Kd of 5 x 10(-10) M and approximately 100 binding sites/cell. Reverse transcriptase-polymerase chain reaction demonstrated that IEC-6 cells as well as epithelial cells from each segment of the rat intestine (duodenum, jejunum, ileum, and colon) contained ER mRNA of the sequence determined from rat uterus. Estradiol was shown to stimulate IEC-6 cell c-fos mRNA content rapidly and transiently in a manner analogous to that which has been previously demonstrated for other estrogen-responsive tissues. These data demonstrate that intestinal epithelial cells contain ER capable of regulating gene transcription and provide the basis for future studies designed to elucidate the role of estrogens in the regulation of intestinal epithelial function and pathophysiology. PMID- 8419142 TI - Interleukin involvement in anterior pituitary cell growth regulation: effects of IL-2 and IL-6. AB - The pituitary gland plays a central role in the interactions between the immune and neuroendocrine systems. The expression of receptors for interleukin-1 (IL-1), IL-2, and IL-6 and the intrinsic production of these ILs by pituitary cells have been described. Previous studies have focused on the way cytokines influence hormone secretion. We have determined whether, in addition to these effects, ILs could affect pituitary cell proliferation. In GH3 cells, both IL-2 (1-100 U/ml) and IL-6 (10-500 U/ml) significantly stimulated [3H]thymidine incorporation and cell count. In contrast, inhibitory effects of both IL-2 and IL-6 at the same concentrations were observed on normal rat anterior pituitary cell growth. This finding was clearly evident when cells were cultured in Minimum Essential Medium D-valine medium, a condition that results in cultures virtually free of fibroblasts. Autoradiographic studies confirmed that [3H]thymidine was only incorporated in the nucleus of nonfibroblastic pituitary cells. No direct correlation between the effects of IL-2 and IL-6 on cell growth and hormone secretion was apparent. By immunofluorescence, we observed IL-2 receptor expression on GH3 cells and, for the normal rat cultures, a high percentage of PRL-secreting and a lower percentage of GH-producing cells expressing IL-2 receptors, providing new evidence for a direct site of action of IL-2 on pituitary cells. Considering that uncontrolled division of cells may result from either excessive growth stimulation or deficient growth inhibition, the regulation of pituitary cell growth by IL-2 and IL-6 together with their intrinsic pituitary production could be of potential importance in pituitary adenoma pathogenesis. PMID- 8419143 TI - The N-acetylglucosamine-specific receptor of the thyroid: purification, further characterization, and expression patterns on normal and pathological glands. AB - The N-acetylglucosamine receptor of the thyroid has been putatively described as both a prohormonal receptor that could play a role in the intrafollicular retention of immature thyroglobulin and a vectorial conveyor of these immature molecules to the iodination site. To further characterize this receptor, we have developed a purification procedure yielding nanomolar amounts of N acetylglucosamine receptor. This thyroid lectin appeared to have an isoelectric point near 5.2 and to be composed of 51-kilodalton monomers with no Asn-linked glycoconjugates. Recognition of the receptor by antipeptide antibodies (Ab/ROV1) raised against a preselected sequence of cation-dependent lectins indicated immunological kinship with the Gal/GalNAc-specific hepatic lectin. Affinity purified Ab/ROV1 and polyclonal antibodies against the purified receptor (TGRD Ab) were used to study the location and expression pattern of the receptor on animal and human thyroid tissue. On porcine slices, positive labeling was observed in various intracellular vesicular compartments with both antibodies and was particularly intense in the apical membrane and subapical compartments. The same pattern was observed in normal human thyroid. In contrast, the receptor 1) could not be found on epithelial cells from thyroid papillary carcinoma; 2) was abundant, but concentrated in the subnuclear region of the thyrocytes in adenomatous goiter; and 3) was almost exclusively located at the basolateral membrane in follicular carcinoma as well as in thyrocytes from glands treated with antithyroid drug before surgery. These observations indicate that expression of the N-acetylglucosamine receptor is characteristic of the fully differentiated phenotype, and its potential function as a thyroglobulin conveyor back to the lumen would be either impaired or abolished in some disease processes. PMID- 8419144 TI - Reduced gonadotropin-releasing hormone gene expression with fasting in the male rat brain. AB - Fasting causes a decrease in serum gonadotropin and testosterone (T) levels in the male rat. We hypothesized that this fasting-induced decrease in serum gonadotropins is due to reduced GnRH gene expression. PreproGnRH mRNA (GnRH mRNA) and serum gonadotropin and T levels by RIA were compared in 90 day old male Wistar rats fed ad lib or fasted for 60 hours (n = 8/group). GnRH mRNA was quantitated by in situ hybridization and computerized image analysis in 18 anatomically matched 20 microns coronal sections from the medial preoptic area (MPOA) and diagonal band of Broca (DBB), using a 35S-labeled 48 base oligodeoxynucleotide probe complementary to rat GnRH mRNA. LH and FSH levels were lower in fasted vs fed rats (LH: 0.17 +/- 0.03 vs 0.36 +/- 0.06 ng/ml, p < 0.01; FSH: 6.7 +/- 0.5 vs 8.7 +/- 0.6 ng/ml, p < 0.05). T levels were also decreased in fasted (1.34 +/- 0.36 ng/ml) vs fed (2.21 +/- 0.30 ng/ml) rats, although this was not statistically significant (p = 0.08). In both the MPOA and DBB, the number of neurons expressing GnRH was lower in fasted vs fed rats (10.6 +/- 0.4 vs 15.8 +/- 1.2 cells/section, p < 0.001), while cellular GnRH mRNA content was unchanged with fasting (83 +/- 1 vs 80 +/- 2 grains/cell). These data support the hypothesis that in the male Wistar rat, decreased gonadotropin secretion with fasting is due, at least in part, to a reduction in GnRH gene expression. PMID- 8419145 TI - The regulation by growth hormone of lipoprotein lipase gene expression is mediated by c-fos protooncogene. AB - GH has been previously shown in Ob1771 adipose cells to activate transiently the expression of c-fos gene by a protein kinase-C-dependent pathway and to modulate, at last in part by a protein kinase-C-dependent pathway, the expression of the lipoprotein lipase (LPL) gene. In Ob1771 cells exposed to GH, under conditions where protein synthesis is inhibited by cycloheximide, the modulation of LPL gene expression is prevented, suggesting that synthesis of trans-acting factor(s) is required to modulate LPL gene expression. The present results indicate the involvement of c-Fos protein in this modulation; this involvement is supported by various lines of evidence: 1) upon GH stimulation, the increase in c-fos mRNA content is followed by the emergence of c-Fos protein within the nucleus, and this emergence precedes the increase in LPL mRNA content; 2) in GH-treated Ob1771 cells, exposure to antisense sof oligonucleotides abolishes the synthesis of c Fos protein; and 3) at the same time, the increase in LPL mRNA content and LPL activity does not occur, whereas sense fos oligonucleotides show no effect. It is concluded that c-Fos protein plays an intermediary role in the modulation of LPL gene expression by GH. PMID- 8419146 TI - Measurement of periimplantational relaxin concentrations in the macaque using a homologous assay. AB - Circulating relaxin concentrations in the human rise in the late luteal phase and increase further in response to increasing circulating CG concentrations immediately after implantation. Similar events have not been documented in the laboratory macaque because of the lack of sensitivity of heterologous assay systems. A homologous enzyme-linked immunosorbent assay for authentic macaque relaxin was developed and validated. Using this enzyme-linked immunosorbent assay, relaxin concentrations were measured in peripheral and ovarian venous blood collected from cynomolgus and rhesus macaques. Relaxin concentrations rose in the late luteal phase of nonconceptive menstrual cycles in cynomolgus macaques, but it was not detected at other times in the cycle. In conceptive cycles, relaxin concentrations rose rapidly in close association with the appearance of mCG 13-14 days after mating. Pregnant rhesus macaques also had elevated relaxin concentrations in blood samples collected on days 15-17 postbreeding. Relaxin concentrations disappeared immediately after luteectomy or ablation of the trophoblast by either surgery or administration of methotrexate. The rise of relaxin paralleled the rise of mCG until 20-25 days postbreeding, while progesterone concentrations declined during this same time period. The lack of correlation between relaxin and progesterone secretion profiles suggests that either the cellular origins or the intracellular mechanisms promoting the secretion of these hormones are different. The periimplantational profile of serum relaxin in macaques was similar to the profile of relaxin observed during early human pregnancy. PMID- 8419147 TI - Characterization of the 1,25-(OH)2D3-induced inhibition of bone nodule formation in long-term cultures of fetal rat calvaria cells. AB - We investigated the effects of 1,25-dihydroxyvitamin D3[1,25-(OH)2D3], on osteoprogenitor cell differentiation and bone nodule formation at various stages of differentiation by evaluating the effects on long term cultures of fetal rat calvaria (RC) cells. RC cells were plated at 3 x 10(4) cells/35-mm dish in alpha minimal essential medium containing 15% fetal bovine serum, ascorbic acid, and beta-glycerophosphate (beta-GP), conditions under which bone nodules form. 1,25 (OH)2D3 inhibited bone nodule formation in a dose-dependent manner with total inhibition occurring at 1-10 nM and half-maximal inhibition occurring at approximately 0.06 nM. 1,25-(OH)2D3 also significantly stimulated RC cell growth in a dose-dependent manner in both the presence and absence of ascorbic acid. Addition of 1 nM 1,25-(OH)2D3 at different times after the start of culture inhibited nodule formation when added before and up to the early multilayering stage (up to day 11 of culture), but had no effect on nodule number when added later. When 1,25-(OH)2D3 was added at the start of the culture period and removed at the early multilayering stage, nodule formation was also inhibited. Pulses of 48-h duration also inhibited nodule formation, with maximal effect occurring between days 3 and 11. Thus, 1,25-(OH)2D3 inhibited osteoprogenitor cell differentiation during the earlier stages of culture before visible bone nodule formation occurred and the effect was not reversible upon removal of 1,25 (OH)2D3. In cultures grown to the multilayering stage in medium without ascorbic acid and beta-GP and then changed to medium with ascorbic acid and beta-GP, 1,25 (OH)2D3 inhibited when present before, but not after, the addition of ascorbic acid and beta-GP. Two other vitamin D3 metabolites, 24,25-dihydroxyvitamin D3 [24,25-(OH)2D3] and 1,24,25-trihydroxyvitamin D3 [1,24,25-(OH)3D3] had inhibitory effects similar to 1,25-(OH)2D3. The effects were dose dependent for each metabolite tested and correlated with the biological effectiveness of these metabolites in other systems: i.e. 1,25-(OH)2D3 was more effective than 1,24,25 (OH)3D3 which in turn was more effective than 24,25-(OH)2D3. The data show that 1,25-(OH)2D3 inhibits osteoprogenitor cell differentiation at an early stage and at a time during which cell growth is stimulated. PMID- 8419148 TI - Alteration of phosphotyrosine phosphatase activity in tissues from diabetic and pregnant rats. AB - Membrane-associated tyrosine phosphatase activities were studied in two distinct states of insulin resistance: diabetes and pregnancy. Using a novel immunoenzymatic assay with intact insulin-like growth factor-I (IGF-I) and insulin receptors as substrates, we show that phosphotyrosine-protein phosphatases (PTP-ases) from normal rat tissues induce a decrease in tyrosine phosphorylation of both receptors. Membrane fractions from kidney, brain, and liver contain the highest PTP-ase activity toward the insulin receptor. After 20 day streptozotocin-induced diabetes, PTP-ase activities are increased by 70% in the placenta, reduced by 40-50% in liver and skeletal muscle, and remained unchanged in the nonclassical insulin target tissues, kidney and brain. In general, the dephosphorylation of IGF-I receptor follows a pattern similar to that of insulin receptor except in red skeletal muscle in which it is not modified. Pregnancy also induces alterations of liver PTP-ases similar to those elicited by diabetes with a 50% reduction of insulin and IGF-I receptor dephosphorylation. This effect of pregnancy is further potentiated by diabetes. The alterations in the activity of hepatic PTP-ases from diabetic and pregnant rats are associated with a decreased autophosphorylation of the insulin receptor, suggesting that the diminution of phosphatase activity might be associated to the state of receptor phosphorylation and activation. Our data demonstrate that alterations of PTP-ases in insulin target tissues are found in two insulin resistant states, one characterized by hyperinsulinemia, pregnancy and one by insulinopenia, streptozotocin-diabetes. These observations suggest a possible relationship between the defective activity of receptor tyrosine kinases and membrane-associated phosphatases from insulin responsive tissues. PMID- 8419149 TI - Coculturing posterior pituitary and GH3 cells: dramatic stimulation of prolactin gene expression. AB - Recent evidence suggests that the posterior pituitary (PP), also called the neurointermediate lobe, regulates PRL release. We previously reported that cocultures of anterior pituitary and PP cells resulted in a 2- to 3-fold increase in PRL content and release. For this study we chose GH3 cells (a somatomammotroph tumor cell line) to determine whether coculturing GH3 with PP cells: 1) stimulates the release and cell content of PRL as compared with GH; 2) increases GH3 cell proliferation; and 3) affects PRL messenger RNA (mRNA) levels. Exp 1. GH3 cells (25,000 cells per well; 25K) were cocultured with PP cells (0K, 12.5K, or 25K) from male rats in serum-free media for 1, 2, 4, and 7 days; hormones were measured by RIA. Coculturing resulted in 5- to 10-fold increases in both media and cell PRL that were linear with time and dependent on the number of PP cells. In contrast, media GH increased only 1.5- to 2-fold, and GH cell content reduced by 75%. Exp 2. GH3, PP, and GH3 + PP cells were cultured for 1, 2, and 4 days and then incubated with [3H]thymidine for 5 h. The incorporation of [3H]thymidine in GH3 cells remained constant over time and showed a small, early increase in cocultures. In contrast, incubation of PP cells alone resulted in a 50- to 60 fold rise in [3H]thymidine incorporation from days 1-4 in culture. Exp 3. Cytoplasmic mRNA was determined by slot blot hybridization with 32P-labeled complementary DNA probes for PRL and GH. After coculturing 25K GH3 cells with 12.5K and 25K PP cells for 4 days, PRL mRNA levels increased 15- and 30-fold, respectively, whereas GH mRNA levels rose less than 2-fold. Neither PRL nor GH mRNA were detected in PP cells. CONCLUSIONS: 1) coculturing GH3 with PP cells dramatically stimulates PRL gene expression, synthesis, and release; 2) this response is specific for PRL, has little effect on GH, and is not due to increased GH3 cell proliferation; and 3) we speculate that a subpopulation of intermediate lobe cells, possibly with a proliferative capacity, is responsible for inducing these effects. PMID- 8419150 TI - Structural basis of the mutagenicity of heterocyclic amines formed during the cooking processes. AB - A data base consisting of 61 heterocyclic amines formed during food preparation and their des-amino analogs were subjected to structure-activity analysis using the CASE method, a structural activity relational expert system. The program identified the major structural determinants associated with mutagenic activity or lack thereof. The structures identified as contributing to the probability of activity as well as those associated with mutagenic potency were highly predictive of molecules not in the learning set. The major structural determinant, the aromatic amino moiety, and quantum mechanical calculations revealed that the mutagenic potency associated with this functionality derived from their contribution to the energy of the Lowest Unoccupied Molecular Orbital (LUMO). PMID- 8419151 TI - The Assessment of Mutagenicity. Health Protection Branch Mutagenicity Guidelines. Health Protection Branch Genotoxicity Committee, Department of National Health and Welfare, Canada. PMID- 8419152 TI - Japanese guidelines for mutagenicity testing. Ministry of Agriculture, Forestry and Fisheries; Ministry of Health and Welfare; Ministry of Labor. AB - Several Japanese agencies are required to perform mutagenicity tests according to regulatory guidelines. Although each agency's guidelines address a specific purpose, the experimental principles behind them are similar, and general methodological recommendations have been issued by the Ministry of Agriculture, Forestry, and Fisheries [1985]; Ministry of Health and Welfare [1990]; Ministry of Labor [1991]; and Ministry of Health and Welfare [1992]. Four major guidelines for mutagenicity testing in Japan and some amendments are briefly introduced. In addition, several procedures in Japanese guidelines that differ from those of other countries or organizations are discussed. PMID- 8419153 TI - Mutagenicity test schemes and guidelines: U.S. EPA Office of Pollution Prevention and Toxics and Office of Pesticide Programs. AB - New requirements for chemicals subject to mutagenicity testing from the U.S. Environmental Protection Agency (USEPA) are discussed. Also detailed are two categories in the 1986 Mutagenicity Risk Assessment Guidelines. PMID- 8419154 TI - Mutagenicity of peroxyacetyl nitrate (PAN) in vivo: tests for somatic mutations and chromosomal aberrations. AB - A series of experiments was conducted in which Chinese hamsters inhaled PAN, an ubiquitous pollutant that is present in the atmosphere at concentrations that are as high as, or higher than, other known genotoxic agents. The animals were exposed to PAN in air at concentrations of approximately 3 ppm for up to 1 month and then examined for somatic mutations and chromosomal aberrations. Mutations were assayed by measuring the frequency of thioguanine-resistant lung fibroblasts (isolated de novo and cultured). Chromosomal aberrations were assayed by measuring the frequency of micronuclei in either the bone marrow (polychromatic erythrocytes) or the lungs (binucleate lung fibroblasts cultured in the presence of cytochalasin B). The results for the test animals were compared to those from animals exposed similarly, but without PAN. Although in each experiment the mutation frequencies for the test animals were higher than the corresponding controls, the mutation frequencies were not significantly different from the concurrent negative controls (P > .05) or the historical controls, except for experiment C. In experiment C, there was a significant regression of mutation frequency versus dose (P < 0.001) if all of the historical controls for pooled animals are included at zero dose. No reproducible evidence of chromosomal breakage was found in either lung or bone marrow. Thus, although PAN has been found to be a bacterial mutagen, we did not find statistically significant evidence of mutagenicity in vivo. The toxicity of PAN limited the exposure concentration that could be used. When all of the PAN data were used, the best estimate of the mutagenic potency proved to be comparable to that of ethylene dibromide, a carcinogenic atmospheric pollutant. PMID- 8419155 TI - Elevation of sister chromatid exchange frequency in transformed human fibroblasts following exposure to widely used aminoglycosides. AB - Aminoglycosides are a class of antibiotics that interfere with protein translation. Geneticin and hygromycin are two such agents, which have been shown to exhibit highly toxic effects in mammalian cells. Cloned bacterial genes, which inactivate these antibiotics, have facilitated the establishment of dominant selection systems, which are widely used in eukaryotic molecular genetics. We have examined the effect of aminoglycosides on the sister chromatid exchange (SCE) frequency in transformed human fibroblast cell lines. Geneticin and hygromycin were both found to increase SCE frequency in all cell lines examined, including a cell line derived from a patient with Bloom syndrome, a disorder exhibiting an elevated spontaneous SCE frequency. Induction was seen to occur in a dose-responsive manner and was also observed in cells expressing the resistance genes that inactivate the cellular toxicity of these antibiotics. The implications of these findings for somatic cell genetics and for human gene therapy protocols are discussed. PMID- 8419156 TI - Incorporation of a micronucleus study into a developmental toxicology and pharmacokinetic study of L-selenomethionine in nonhuman primates. AB - Concomitant to a developmental toxicology study of selenium in long-tailed macaques (Macaca fascicularis), a transplacental bone marrow micronucleus assay was conducted in the fetuses of treated animals. Selenium was administered as L selenomethionine by nasogastric intubation at 0, 150 or 300 micrograms/kg-day to pregnant macaques daily throughout organogenesis (gestation days 20-50). Pregnancy was terminated on gestation day 100 +/- 2 and fetuses were obtained by hysterotomy. Selenium concentrations in maternal blood were monitored throughout pregnancy and selenium concentrations in fetal blood were measured at hysterotomy. Maternal circulating selenium did not exceed 4 ppm in plasma or 3.7 ppm in erythrocytes. Selenium in cord blood was < or = 0.1 ppm in plasma and < or = 1.1 ppm in erythrocytes at 300 micrograms/kg-day. Fetal bone marrow smears were prepared from the humerus and micronucleated polychromatic erythrocytes were scored. No increase of micronucleus frequency was detected in any dose group, although signs of maternal selenosis were obvious. This finding is compared to the previous observation that micronuclei were induced in the bone marrow of adult nonpregnant macaques treated at 600 micrograms/kg-day, a lethal dose yielding blood selenium levels to 7.3 ppm in plasma and 5.7 ppm in erythrocytes after 15 days of daily treatment, when death occurred. These data demonstrate that measurement of circulating xenobiotics can be useful for the interpretation of genetic toxicology results. PMID- 8419157 TI - Genetic toxicology testing requirements: official and unofficial views from Europe. AB - This work presents genetic toxicity testing requirements in three industry sectors; pharmaceuticals, pesticides, and industrial chemicals. PMID- 8419158 TI - In vitro cytotoxic and cell transforming activities exerted by the pesticides cyanazine, dithianon, diflubenzuron, procymidone, and vinclozolin on BALB/c 3T3 cells. AB - Cytotoxic and cell transforming activities of the pesticides cyanazine, diflubenzuron, dithianon, procymidone, and vinclozolin were investigated in vitro by utilizing the BALB/c 3T3 cell transformation test performed in the presence or in the absence of S-9 mix as an exogenous bioactivation system for the chemicals. All the assayed pesticides were cytotoxic in the absence of S-9 mix, whereas only dithianon exerted cytotoxic effects in the presence of metabolic activation. All the chemicals tested did induce BALB/c 3T3 cell transformation, to a various extent, in the absence of S-9 mix. Cell transforming ability of cyanazine and diflubenzuron was not detectable in the presence of S-9. PMID- 8419159 TI - Selective aflatoxin B1-induced sister chromatid exchanges and cytotoxicity in differentiating B and T lymphocytes in vivo. AB - The purpose of this study was to assess the genotoxic and cytotoxic effects of the fungal metabolite aflatoxin B1 (AfB1) on the developing immune system of the chick embryo, a model in vivo system. Of particular interest was the assessment of AfB1-mediated selective toxicity toward developing B lymphocytes as compared to T lymphocytes. In vivo bromodeoxyuridine (BrdU) labelling of DNA was used to detect the induction of sister chromatid exchanges (SCE) in lymphocytes and to assess the progression of these cells through successive cell cycles. Cytotoxicity was also assessed by studying the entrance and maintenance of cells in mitosis (mitotic index). Graded doses of AfB1 (1.09-17.4 micrograms/g embryo) were applied to chick embryos at 18 days of incubation (DI). Embryos also received two doses of BrdU at 3 mg/200 microliters (3 hr apart) to provide continuous labelling of B and T lymphocyte replicating DNA. B and T lymphocytes were harvested 20 hr post-AfB1/BrdU exposure from the bursa and thymus, respectively, and were processed for cytogenetic analyses. AfB1 induced dose related increases in SCE in B lymphocytes; this induction was 6- to 8-fold that of controls at the higher doses tested. AfB1-mediated induction of SCE in T cells was just 2-fold that of controls at the highest dose tested. AfB1 reduced the progression of B cells and to a lesser extent T cells through successive rounds of replication. Furthermore, AfB1 dramatically reduced the mitotic index of B cells but not of T cells. These data indicate both selective genotoxicity and cytotoxicity of AfB1 toward B cells in the late stage embryo. PMID- 8419160 TI - Mutagenicity of the human carcinogen treosulphan, and its hydrolysis product, dl 1,2:3,4-diepoxybutane in mammalian cells. AB - The cytotoxicity and mutagenicity of the human carcinogen, treosulphan, and its hydrolysis product, dl-1,2:3,4-diepoxybutane (DEB), were studied in Chinese hamster ovary, AS52, cells. Treosulphan (0.1-1.0 mM) is toxic and mutagenic at the gpt locus. A strong pH dependence was noted. DEB is cytotoxic and mutagenic at a much lower dose (0.025 mM), but these effects were not affected by pH. The results suggest that the toxic and mutagenic effects of treosulphan are mediated by its hydrolysis product DEB, and that the conversion of treosulphan to DEB is highly pH-dependent. PMID- 8419161 TI - Mechanisms that generate human immunoglobulin diversity operate from the 8th week of gestation in fetal liver. AB - The repertoire of immunoglobulin expressed very early in human development was approached by cloning and sequencing 55 rearranged and 11 germ-line VH transcripts, after amplification by polymerase chain reaction of cDNA libraries derived from two fetal livers at 8 and 13 weeks of gestation. All families with the exception of VH2, were expressed as soon as 8 weeks, with preferential usage of certain germ-line genes. Very few somatic mutations, randomly localized, were identified. By contrast, in a series of clones derived from the same VDJ rearrangement using the VH6 family, extensive mutations had taken place, mostly accumulated in the third complementarity-determining region (CDR3) suggesting that the specialized enzymatic machinery was at hand very early during human development. Some other characteristics of the fetal repertoire also emerged, namely increased usage of JH3 and JH2, as compared to the adult pattern, where JH4 is dominant and reduced length of the D/CDR3 regions. All D gene families were identified, and their usage frequently involved D-D fusions. N diversity was present very early, and increased with age. Identification of germ-line transcripts pertaining to all six VH families including pseudogenes, in the E55 library, revealed a population very different as compared to rearranged gene transcripts. This suggests that a large portion of VH locus is accessible for transcription, bringing no evidence of correlation between preferential rearrangement of a given VH gene and its localization in the locus. PMID- 8419162 TI - Fine specificity and VJ usage of light chains in antibodies to the phosphorylcholine hapten. AB - In the memory response to the phosphorylcholine hapten (PC) two major groups of anti-PC antibodies with different fine specificities are elicited. Group I antibodies are mainly PC specific, whereas Group II antibodies are comprised of two specificities directed against the phenyl-PC and the phenyl moiety of the PC hapten. The VL gene usage of 17 monoclonal memory anti-PC antibodies were investigated by Southern blot analysis and nucleotide sequencing. Six out of eight Group I memory PC-specific antibodies used the same VK22-JK5 rearrangement as the major T15 primary response idiotype. One expressed a mutated JK1 and one employed another VK22 gene family member. A shift in specificity from PC (Group I) towards phenyl-PC (Group II) was accompanied with the usage of either VK1C-JK1 or VK1A-JK5 rearrangements. The phenyl-specific Group II antibodies expressed the V lambda 1-J lambda 1 L chain rearrangement in combination with VH M141 expressing H chains. In this specific segment of Group II antibodies most mutations were found. Thus four different VL genes were found to contribute to the fine specificity of memory response antibodies to the PC hapten in a clear structure-function relationship. The diversified fine specificity in the memory response derives mainly from the usage of different L chains with particular VJ rearrangements in combination with VH of the dominant initial response clonotype and is not primarily due to mutational events. PMID- 8419163 TI - Protein tyrosine kinases couple the interleukin-2 receptor to p21ras. AB - The T cell growth factor interleukin-2 (IL-2) induces p21ras activation in T lymphocytes. We have previously shown that a protein kinase C (PKC)-mediated pathway for p21ras regulation exists in T cells and that the IL-2 receptor (IL 2R) can couple to p21ras independently of the presence of the PKC pathway for p21ras regulation. Our data show that in conditions where cellular protein tyrosine kinases (PTK) were efficiently down-regulated by pretreatment with the specific PTK inhibitor herbimycin, the IL-2-induced activation of p21ras was blocked. Herbimycin did not inhibit the PKC-mediated pathway for p21ras regulation. Thus, the data indicate that PTK are involved in the coupling of the IL-2R to p21ras. PMID- 8419164 TI - Recognition of shared melanoma antigen by HLA-A2-restricted cytolytic T cell clones derived from human tumor-infiltrating lymphocytes. AB - Three melanoma-specific cytotoxic T lymphocytes (CTL) clones were derived from the tumor-infiltrating lymphocyte (TIL) of human melanoma M17, and were used to study the expression of immunogenic melanoma peptides on allogeneic tumors. Antibody inhibition studies showed that two of these TIL clones were restricted by an HLA-A2 molecule which was identified as A2.1 by gene sequencing. The third CTL clone was not restricted by HLA-A2, but by a B or C HLA antigen. HLA-A2 restricted CTL clones M17-1 and M17-2 lysed 5 and 12 out of 15 HLA-A2+ allogeneic melanomas, respectively. Since they did not lyse autologous Epstein-Barr virus B cells, HLA-A2.1-transfected P815 cells, 13 HLA-A2+ non-melanoma tumor cell lines and 10 HLA-A2- melanomas, these clones appeared specific for melanoma-restricted epitopes presented by the HLA-A2.1 molecule. We then tried to determine why a few HLA-A2+ melanomas were refractory to TIL lysis. By using a combination of flow cytometry analysis, partial cloning and sequencing of their HLA-A2 genes, we show that failure to lyse did not result from low expression or polymorphism of the HLA-A2 molecule, or from deficient expression of the adhesion molecules ICAM-1 and LFA-3 by these melanomas. Taken together, our data confirm at the clonal level the existence of shared melanoma antigens recognized by TIL in the HLA-A2.1 context. They further show that individual peptides derived from these antigens are expressed by a large majority of HLA-A2+ melanomas. Identification of such peptides appears crucial for the future of vaccination therapies. PMID- 8419165 TI - Correlation between cell-mediated immunity and degree of infection in subjects living in an endemic area of schistosomiasis. AB - Cell-mediated immunity to Schistosoma mansoni antigens, unrelated antigens and mitogens was evaluated in 50 subjects with the same degree of exposure to infection living in an endemic area of schistosomiasis. The degree of infection, assessed by the number of eggs/g of stool, was variable in this population (0 5604), suggesting differences in susceptibility to infection. Absence of lymphoproliferative response was observed in 56% of this group, despite having a response to purified protein derivative of tuberculin (PPD) and tetanus toxoid (TT) antigens and to pokeweed mitogen. The 50 subjects were divided into two groups, according to their degree of infection. The lymphoproliferative responses to schistosomula and adult worm antigens in the group with a low degree of infection (< 400 eggs/g of stool) were higher than the ones documented in patients with a high degree of infection (> 400 eggs/g of stool), and these differences were statistically significant (p < 0.001). An inverse correlation between the lymphocyte proliferation in response to S. mansoni antigens and the degree of infection was also observed (p = 0.02), indicating that subjects with a lower degree of infection have a higher lymphoproliferative response to schistosomula and adult worm antigens. No differences in the lymphocyte reactivity to other antigens (PPD and TT) were detected in these groups. An impairment of interferon-gamma in vitro production was observed when the lymphocytes from these subjects were stimulated with S. mansoni adult worm antigen, although they produced gamma interferon in response to phytohemagglutinin. PMID- 8419166 TI - Characterization of four novel epsilon chain mRNA and a comparative analysis of genes for immunoglobulin E in rodents and man. AB - The nucleotide sequence of the 3' region of the epsilon chain gene for human IgE is presented. A comparison of the entire region from 5' of exon C1 to the M2 exon of the mouse, rat and human epsilon chain genes shows that the overall structure of the epsilon chain gene have changed only minimally during the 60-70 million years of evolutionary separation between rodents and man. We have previously shown that a number of rearrangements larger than 10 bp have relatively recently occurred in the C4/M1 intron of the rat or the mouse epsilon chain genes. A majority of these rearrangements were found within or in close proximity to repetitive sequences of Z-DNA-forming potential (CA dinucleotide repeats). The C4/M1 intron has evolved very rapidly, to such an extent that no apparent homology can be detected between rodents and man. Only remnants of the repetitive sequences are present in man, supporting the theory that repetitive sequences having Z-DNA-forming properties may play a role in the evolution of the eucaryote genome by promoting recombinations, leading to a rapid evolutionary drift of sequences in close proximity to these repeats. We report here the characterization of the membrane domains of human IgE and four novel mRNA transcribed from the human epsilon chain locus. The primary structures have been determined by polymerase chain reaction cloning and nucleotide sequence analysis. All five mRNA contain the C3 domain and the membrane exon 2 (M2). Due to frame shifts caused by novel splice sites or novel splice-site combinations, the proteins encoded by three out of these four novel mRNA differ in their carboxy terminal end from the classical secreted or membrane-bound immunoglobulins. Northern blot analysis shows significant levels of at least three out of these four novel mRNA in an IgE-producing human cell line. One of the mRNA encodes a transmembrane-like structure which has characters in common with the transmembrane region of the CD3 components of the T cell receptor complex (CD3 gamma, delta and epsilon). This indicates that IgE-producing B cells possibly have two separate signal-transducing systems. A comparison of the classical membrane anchoring domain of the human & chain with a panel of immunoglobulin membrane domains from fish to higher mammals is presented. A tyrosine and a glutamine residue is found to be conserved between all cytoplasmic domains of all post-switch immunoglobulin classes indicating a functional conservation of these amino acid residues.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419167 TI - A role for immunoglobulin D: interference with tolerance induction. AB - We have studied the induction and maintenance of tolerance in the B lymphocyte compartment. Neonatal and adult transgenic mice which expressed either surface IgM (sIgM) or sIgM and sIgD anti-2,4,6-trinitrophenyl (TNP) were treated with soluble mono- and multivalent forms of TNP-modified carriers. We compared the B cell compartment of mice treated with antigen and of littermates injected with phosphate-buffered saline. Antigen-mediated cross-linking of membrane-bound IgM (sIgM) caused deletion of B cells both in neonatal and adult mice with mu and kappa transgenes. Deletion was the result of apoptosis. In mice that carried an additional delta transgene sIgD interfered with tolerance induction. The stage in which the cells were sensitive to deletion was characterized as a transitional stage between immature (sIgMdull, heat-stable antigenbright, B220dull, sIgD-) and more mature (IgMbright, heat-stable antigendull, B220bright, sIgD-) B cells. Surviving cells were functional as measured by receptor-mediated changes in the intracellular free Ca2+ concentration. We propose that when the immature B cells have reached the final stages of maturation IgM always transmits negative signals in the absence of T cell help. When B cells need to be screened against self reactivity IgM is the only antigen receptor expressed. The presence of sIgD protects resting B cells from deletion and allows them to initiate an effective immune response. PMID- 8419168 TI - T suppressor hybridomas and interleukin-2-dependent lines induced by copolymer 1 or by spinal cord homogenate down-regulate experimental allergic encephalomyelitis. AB - Suppressor T (Ts) hybridomas and interleukin-2-dependent T cell lines were established from spleens of mice, which had been rendered unresponsive to experimental allergic encephalomyelitis (EAE) either by mouse spinal cord homogenate or by the synthetic suppressant copolymer 1 (Cop 1). The Ts hybridoma supernatants and the Ts line cells specifically suppressed the in vitro response to the encephalitogenic myelin basic protein (BP), as indicated by inhibition of both the proliferation and interleukin-2-secretion responses of a BP-specific T cell line. Moreover, these Ts cells prevented the development of actively induced EAE in vivo. All hybridomas and lines were most effective when injected at the time of disease induction, thus suggesting that they operate as effector suppressor cells, and functionally inhibit encephalitogenic responses. The data presented here suggest that the suppressor cells are stimulated by the protective epitopes included in the BP as well as in the Cop 1 molecules and that they play an active role in the regulation of EAE. The generation of Ts lines and hybridomas, which have been induced by Cop 1, establish the specific stimulation of suppressor cells to EAE as a mechanism underlying the therapeutic activity of Cop 1. PMID- 8419169 TI - Inhibition of human immunodeficiency virus infection by the lectin jacalin and by a derived peptide showing a sequence similarity with gp120. AB - Jacalin is a plant lectin known to specifically induce the proliferation of CD4+ T lymphocytes in human. We demonstrate here that jacalin completely blocks human immunodeficiency virus type 1 (HIV-1) in vitro infection of lymphoid cells. Jacalin does not bind the viral envelope glycoprotein gp120. Besides other T cell surface molecules, it interacts with CD4, the high-affinity receptor to HIV. Binding of jacalin to CD4 does not prevent gp120-CD4 interaction and does not inhibit virus binding and syncytia formation. The anti-HIV effect of the native lectin can be reproduced by its separated alpha-subunits. More importantly, we have defined in the alpha-chain of jacalin a 14-amino acid sequence which shows high similarities with a peptide of the second conserved domain of gp120. A synthetic peptide corresponding to this similar stretch also exerts a potent anti HIV effect. This peptide is not mitogenic for peripheral blood mononuclear cells and does not inhibit anti-CD3-induced lymphocyte proliferation. These results make jacalin alpha chain-derived peptide a potentially valuable therapeutic agent for acquired immunodeficiency syndrome. PMID- 8419170 TI - Macrophage colony-stimulating factor gene transfer into tumor cells induces macrophage infiltration but not tumor suppression. AB - In order to analyze the effect of a high local concentration of macrophage colony stimulating factor (M-CSF; CSF-1) on tumor growth, the plasmacytoma cell line J558L was transfected with the human M-CSF gene and injected into syngeneic BALB/c mice. In contrast to the parental tumors, M-CSF transfectants were heavily infiltrated by macrophages as evidenced by immunohistochemistry with antibodies to Mac-1 and Mac-3 and by isolation of the macrophages from the tumor. Nevertheless, tumor growth was only slightly affected by M-CSF and M-CSF producing cells grew as tumor in all cases. The growth retardation of M-CSF producing cells varied depending on the experiment and seemed to be due to an indirect effect because the growth rate of the cells in vitro had not changed upon gene transfer. Attempts to activate the tumor-infiltrating macrophages for tumor suppression by systemic application of interferon-gamma and/or lipopolysaccharide were not successful. Altogether, our results suggest that M CSF is a potent chemoattractant for macrophages in vivo but alone is not sufficient to activate these macrophages for tumoricidal activity. PMID- 8419171 TI - Regulation of T helper cell cytokine expression: functional dichotomy of antigen presenting cells. AB - A bias to either cell-mediated or antibody-mediated effector mechanisms is induced in an immune response against a pathogen, if activated T helper cells (Th) predominantly express Th1 [interleukin (IL)-2, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-beta] or Th2 (IL-4, IL-5, IL-6 and IL-10) cytokines. Here we provide evidence that, due to the capability to secrete IL-1, macrophages, but not B cells, as antigen-presenting cells (APC) induce production of IFN-gamma in resting Th cells. Normal murine splenic Th cells were activated in vitro with the superantigen Staphylococcus aureus enterotoxin B (SEB) presented by macrophages as compared to other APC from murine spleen. As determined by immunofluorescence, Th cells producing IL-2 but almost none producing IL-4 and IL-5 are generated, irrespective of the type of APC. Generation of IFN-gamma-producing Th cells is largely dependent on presentation of SEB by macrophages. The requirement for macrophages, however, is overcome if IL-1 is provided. Expression of IFN-gamma by Th cells is not induced, if production of IL-1 by macrophages is inhibited by IL-10. Our results suggest a functional dichotomy of APC: normal resting Th cells differentiate into IL-2 and IFN-gamma secreting cells (Th1 cells) if antigen is presented by macrophages, whereas presentation by B cells generates Th cells secreting IL-2, which might differentiate into Th2 cells upon re-stimulation. PMID- 8419172 TI - Development of T cell receptor alpha beta-bearing T cells in the submersion organ culture of murine fetal thymus at high oxygen concentration. AB - Organ culture (OC) of murine fetal thymuses on a membrane filter floating on the medium has been used as an important strategy in studies of the mechanism of T cell development. On the other hand, the submersion organ culture (S-OC) system, in which fetal thymus lobes are submerged in the culture medium, is not popularly used because the growth of T cells is much lower than that in OC at the air medium border (AMB-OC). In the present work, we tried to culture the fetal thymuses in the S-OC system at 5% CO2 and various concentrations of O2. We found that in the environment containing 5% O2, gamma delta T cells were selectively generated, though the cell recovery was less than one-tenth of that seen in AMB OC. Generation of gamma delta T cells was hardly affected by increasing the O2 concentration. In marked contrast, the development of alpha beta T cells was highly dependent upon the O2 concentration. In the S-OC at more than 60% O2, differentiation and growth of T cells, as determined in terms of recovered cell number, CD4 vs. CD8 profile and predominance of alpha beta lineage, were comparable to those observed in AMB-OC. It was further shown that clonal deletion of V beta 6+ T cells occurred in high O2 S-OC of CBA/J (Mls-1a) but not C3H/HeJ (Mls-1b) mice, the extent of the deletion being low but comparable with that seen in AMB-OC. These results indicated that the fetal thymus microenvironment was potentially capable of supporting the development of both gamma delta and alpha beta T cells, and that skewing to one of these lineages was determined by an additional factor(s) such as the concentration of O2 dissolved in the medium. PMID- 8419173 TI - Optimization of primers for cloning libraries of mouse immunoglobulin genes using the polymerase chain reaction. AB - We have optimized primers for cloning libraries of murine heavy and light chain variable regions using the polymerase chain reaction. Since we are interested in cloning murine Fab fragments for expression in bacterial cells, the heavy chain primers were designed to clone Fd fragments comprising the heavy chain variable domain and the first domain of the IgG constant region. The light chain primers were designed to clone the entire murine kappa chain. Using ten degenerate 5' primers and a degenerate 3' primer to amplify murine Fd and seven degenerate 5' primers with a single 3' primer to amplify kappa chains, a diverse repertoire of mouse variable regions was cloned from mouse spleens. PMID- 8419174 TI - CD8 is needed for positive selection but differentially required for negative selection of T cells during thymic ontogeny. AB - During thymic development, immature thymocytes expressing major histocompatibility complex (MHC) class I-restricted T cell receptors (TcR) differentiate into CD8+ T cells with cytolytic functions. To evaluate the role of CD8 in positive and negative selection during thymic ontogeny, mice rendered CD8 null by gene targeting were bred with three lines of transgenic mice expressing unique MHC class I-restricted TcR. In all three instances CD8 was required for positive selection of MHC class I-restricted transgenic T cells. The efficiency of positive selection decreased in accordance with a reduced level of CD8 expression on thymocytes. Surprisingly, there was a differential requirement for CD8 expression in negative selection of MHC class I-restricted thymocytes, depending on the antigen specificity of TcR. These observations show that CD8 is essential for positive selection but is differentially required for negative selection of MHC class I-restricted T cells. Thus thymic selection, at least for negative selection, can occur in the absence of the CD8 accessory molecule. PMID- 8419175 TI - Leishmania mexicana promastigotes induce cytotoxic T lymphocytes in vivo that do not recognize infected macrophages. AB - The question is addressed whether antigens of Leishmania, a parasite residing in the endosomal compartment of macrophages, can be presented in the context of major histocompatibility complex class I molecules. We used E. coli beta galactosidase as a model antigen which can be expressed in high levels in L. mexicana promastigotes (L. mexicana-gal). Infection of BALB/c mice with L. mexicana-gal induces beta-galactosidase-specific cytotoxic T cells (CTL), which can be isolated using a beta-galactosidase-expressing mastocytoma line as an antigen-presenting cell. These CTL recognize epitopes of beta-galactosidase in the context of H-2Kd; however, they do not recognize L. mexicana-gal-infected macrophages even after killing of the intracellular amastigotes by drug treatment or macrophage activation by lymphokines, although class I-peptide interaction and the presentation of endogenously produced antigens is normal. It is concluded that parasite antigens can induce a CTL response in vivo but that these CTL cannot recognize infected macrophages because the relevant epitopes cannot gain access to class I molecules. The effect of priming in vivo may be explained by the well-known but ill-understood phenomenon of cross-priming. PMID- 8419176 TI - Signal joint of immunoglobulin V lambda 1-J lambda and novel joints of chimeric V pseudogenes on extrachromosomal circular DNA from chicken bursa. AB - We isolated extrachromosomal circular DNA from the bursa of 18-day chick embryos and cloned their BamHI fragments into a phage vector. We found examples of the signal joint fused by V lambda 1-J lambda rearrangement and the sequences homologous to V lambda 1 segments that showed: (1) clustered V pseudogene (psi V) germ-line segments containing new psi V26, (2) a head-to-tail duplication of psi V13-psi V12 region and (3) chimeric structures composed of 5'-V lambda 1 and 3' psi V segments. Both intrachromosomal tandem duplications and extrachromosomal circles may be generated by unscheduled DNA synthesis and recombination. The chimeric structure of V lambda 1 joined with upstream psi V segments suggests the involvement of V gene replacement for somatic diversification of the immunoglobulin genes in addition to a mechanism of segmental gene conversion. PMID- 8419177 TI - Functional double-negative T cells in the periphery express T cell receptor V beta gene products that cause deletion of single-positive T cells. AB - A proportion of peripheral T cells lack surface expression of the CD4 or CD8 coreceptor molecules and hence are designated as " double negative" (DN). Most DN T lymphocytes express the gamma/delta T cell receptor (TcR), but a minor fraction of them, in both humans and mice, express the alpha/beta TcR. Whereas alpha/beta+ DN T lymphocytes are infrequent (< 1%) in conventional lymphoid organs (spleen, blood, lymph node), they account for two-thirds of the T cells residing in adult bone marrow. Analysis of the TcR V beta repertoire expressed by peripheral DN T cells revealed a high frequency of cells bearing autoreactive TcR that cause deletion of "single-positive" (SP) (CD4+CD8-or CD4-CD8+) T cells. Peripheral DN cells thus represent a cell type that is relatively resistant to clonal deletion. Furthermore, such cells have not been inactivated (anergized) in vivo since they proliferate and secrete interleukins in response to cross-linking by monoclonal antibodies specific for these V beta gene products that are deleted in SPT cells. These results might help to understand the association of peripheral expansion of DN cells and development of autoimmune diseases. PMID- 8419178 TI - The role of natural antibodies and ABO (H) blood groups in transplantation of human lymphoid cells into mice. AB - Recently, evidence was presented that natural antibodies (NAb) are a crucial barrier to human cellular engraftment in severely immunosuppressed normal mice (Eur. J. Immunol. 1992. 22: 197.). In this report we show that normal mouse serum contains low titers of NAb against human cells of blood groups type O (H) and B and high titers against human cells of blood group A. Accordingly, human peripheral blood leukocytes (PBL) of group O (H) and B donors could be grafted successfully into normal BCBA mice (H-2b/k) following irradiation with high dose total body irradiation (TBI). PBL of blood group type A donors did not engraft in normal mice but could be transplanted without difficulty in B cell-deficient CBA/N mice which lack NAb, after conditioning with high dose TBI. Treatment of lethally irradiated normal BCBA mice with cobra venom factor (COF), which eliminates the third factor of complement, and liposomes containing dichloromethylene diphosphate (Cl2DMP), which eliminates macrophages, resulted in engraftment of human blood group type A PBL. This implies that the NAb barrier for discordant xenogeneic cell transplantation can be abrogated. A method utilizing directly labeled probes and flow cytometry is described for the quantitation in mouse serum of NAb, reacting with human cells. Using sera of H 2b/k mice we show that murine NAb react with human stem cells, granulocytes, lymphocytes and monocytes of blood group A and only weakly with similar cells from blood group O (H) and B donors. Sera of H-2b, H-2d and H-2k mice of different ages and microflora possess NAb against human erythrocytes of blood group type A and occasionally demonstrate weak titers against erythrocytes of blood groups B and O (H) and the Rhesus factor. PMID- 8419179 TI - Reconstitution of C.B-17 scid mice with BALB/c T cells initiates a T helper type 1 response and renders them capable of healing Leishmania major infection. AB - C.B-17 scid mice, which were found to be very susceptible to infection with Leishmania major, were reconstituted with various doses of T cells, T plus B cells or unfractionated spleen cells from nonhealer BALB/c mice. All reconstitution protocols, except for the transfer of very high numbers of BALB/c spleen cells, led to a spontaneously healing infection and resistance to reinfection, rather than the lethal, nonhealing infection typical of BALB/c mice. These healing responses were associated with a strong T helper 1 (Th1)-like response characterized by delayed-type hypersensitivity (DTH) responsiveness, but no elevation of serum IgE, and by the production of high levels of interferon gamma (IFN-gamma), but no interleukin-4 (IL-4) by lymph node and spleen cells after restimulation with antigen in vitro. The development of this Th1 response from BALB/c Th cells requires IFN-gamma during the initial infection period. Treatment of scid mice with a single injection of neutralizing anti-IFN-gamma antibody prior to infection and reconstitution prevented healing and permitted the development of a Th-2 like response as indicated by elevated serum IgE, but no DTH, and by the production of IL-4, but very little IFN-gamma, after antigen stimulation in vitro. As few as 10(4) transferred T cells led to a Th1-like response, suggesting that the IFN-gamma is of host rather than donor origin. The transfer of very high numbers (7.5 x 10(7)) of BALB/c spleen cells overcame the effects of the IFN-gamma and led to the nonhealing infection and cytokine pattern characteristic of BALB/c mice. The enrichment or depletion of B cells from the transferred T cells had no measurable effect upon the development of a healing response in reconstituted scid mice. PMID- 8419180 TI - The effects of a monoclonal antibody to interferon-gamma on experimental autoimmune thyroiditis (EAT): prevention of disease and decrease of EAT-specific T cells. AB - CBA/J mice immunized with thyroglobulin (Tg) develop an experimental autoimmune thyroiditis (EAT) with lymphocytic infiltration of the thyroid glands, autoantibodies to Tg and occurrence of EAT-specific T cells. When these mice were treated for 4 weeks after immunization with 1 mg/week of a monoclonal antibody (mAb) that neutralizes the activity of interferon-gamma (IFN) a beneficial effect on the onset of EAT was observed. Characteristic features of EAT were significantly reduced, including the lymphocytic infiltrations of the thyroid glands and the serum levels of autoantibodies to Tg. Moreover, in lymphoid organs, mAb to IFN-gamma significantly reduced the percentages of Tg-specific CD8+ cells, labeled by the anti-clonotypic mAb AG7. These Tg-specific T cells seem responsible for thyroid damages and disease development, since EAT was simultaneously abrogated. These results show that IFN-gamma plays an essential role in the pathophysiology of EAT and suggest the possibility to treat autoimmune thyroid diseases with mAb to IFN-gamma or drugs able to antagonize the production and/or the action of this cytokine. PMID- 8419181 TI - Charge-dependent binding of granzyme A (MTSP-1) to basement membranes. AB - The murine T cell-associated serine proteinase granzyme A [also termed Murine T cell-associated serine proteinase-1 (MTSP-1), or SE-1] expresses optimal enzymatic activity under extracellular milieu conditions. It degrades a variety of proteins that are constituents of basement membranes. Granzyme A is harbored within intracellular storage granules from which its release can be induced by appropriate ligand binding to extracellular matrix receptors of T cells. Secreted granzyme A has, therefore, been implicated in the degradation of extracellular matrix barriers during T cell migration. Here we show that granzyme A binds to natural basement membranes in a charge-dependent manner. Binding of granzyme A to charged surfaces protects if from inhibition by natural high molecular weight inhibitors. The interaction of granzyme A with in vitro-produced extracellular matrices liberates basic fibroblast growth factor, which is bound to negatively charged heparan sulfate glycosaminoglycans of the extracellular matrix. We propose that the charge-dependent interaction of granzyme A with basement membranes has multiple, biologically relevant consequences. PMID- 8419182 TI - Serum modulates mast cell responses to IgE antigen stimulation. AB - Serum induces the expression of the fos and jun gene families, which encode the transcription factor AP-1. Since we previously found that activation of mast cells by IgE-antigen (Ag) induces the mRNA accumulation of c-fos, c-jun, junB and junD proto-oncogenes, we were prompted to investigate whether serum could affect such accumulation in these cells. In addition, we investigated whether serum could modulate inhibition of DNA synthesis in immunologically stimulated mast cells. Mast cells, which were cultured in the presence of fetal calf serum (FCS), were characterized by a high proliferation rate and high accumulation of the mRNA of c-fos, junB and junD proto-oncogenes. After sustained FCS deprivation both DNA synthesis and the level of c-fos mRNA were significantly decreased, as expected, whereas the level of c-jun, junB and junD mRNA were not affected. As opposed to mast cells which were cultured in the presence of FCS, immunological stimulation of FCS-deprived cells resulted in DNA synthesis inhibition and an increase in c fos expression. The results also show that the level of c-fos mRNA was increased by either IgE-Ag or FCS up to a similar level, while these two triggers could not act synergistically to enhance this expression further. Thus, changes in DNA synthesis, induced by FCS, block the ability of the immunological challenge to inhibit mast cell growth and to enhance c-fos mRNA accumulation. PMID- 8419183 TI - Neutrophil attractant/activation protein-1 (interleukin-8) induces in vitro neutrophil migration by haptotactic mechanism. AB - The role of leukocyte migration induced by the substrate-bound attractants is obscured by the inability of standard methods for the study of leukocyte migration to dissociate chemotaxis and haptotaxis, migration in response to soluble and surface-bound gradients of attractants. Here we show that the gradient of neutrophil attractant/activation protein-1 (interleukin-8, NAP-1/IL 8) induces directed in vitro neutrophil migration when bound to the polycarbonate filter. In addition, we suggest that haptotaxis is responsible for neutrophil migration induced by NAP-1/IL-8 in standard Boyden-type chemotaxis assays and, in light of the ability of NAP-1/IL-8 to bind to the endothelial cell surface and the extracellular matrix, could also be of great significance in vivo. PMID- 8419184 TI - NK1.1+ CD4+ CD8- thymocytes with specific lymphokine secretion. AB - CD4+8- or CD4-8+ thymocytes have been regarded as direct progenitors of peripheral T cells. However, recently, we have found a novel NK1.1+ subpopulation with skewed T cell antigen receptor (TcR) V beta family among heat-stable antigen negative (HSA-) CD4+8- thymocytes. In the present study, we show that these NK1.1+ CD4+8- thymocytes, which represent a different lineage from the major NK1.1- CD4+8- thymocytes or CD4+ lymph node T cells, vigorously secrete interleukin (IL)-4 and interferon (IFN)-gamma upon stimulation with immobilized anti-TcR-alpha beta antibody. On the other hand, neither NK1.1- CD4+8- thymocytes nor CD4+ lymph node T cells produced substantial amounts of these lymphokines. A similar pattern of lymphokine secretion was observed with the NK1.1+ CD4+T cells obtained from bone marrow. The present findings elucidate the recent observations that HSA- CD4+8- thymocytes secrete a variety of lymphokines including IFN-gamma, IL-4, IL-5 and IL-10 before the CD4+8- thymocytes are exported from thymus. Our evidence indicates that NK1.1+ CD4+8- thymocytes are totally responsible for the specific lymphokine secretions observed in the HSA- CD4+8- thymocytes. PMID- 8419185 TI - Cytokine regulation of the interleukin-1 receptor antagonist protein in U937 cells. AB - A naturally occurring receptor-level antagonist of interleukin-1 (IRAP or IL-1 ra) has recently been cloned. To determine what stimuli might regulate this inhibitor, cytokines were tested for their effects on the steady-state level of IRAP mRNA in phorbol ester-differentiated U937 cells. The cytokines tested fell into one of three groups: (a) inducers: granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-4, (b) weak inducers (< 2-fold stimulation): [IL-1 alpha, IL 1 beta, and transforming growth factor-beta (TGF-beta)] and (c) cytokines with no effect: (IL-2, platelet-derived growth factor, acidic fibroblast growth factor, basic fibroblast growth factor, epidermal growth factor, granulocyte colony stimulating factor, IL-3, IL-5, IL-6, interferon-gamma, multi-colony stimulating factor, tumor necrosis factor-alpha and IRAP itself. One hundred U/ml of either GM-CSF or IL-4 was the dose inducing peak IRAP mRNA expression; that peak expression occurred 12 h after addition of cytokine. GM-CSF induced a 34 +/- 15 fold increase in IRAP mRNA, and IL-4 induced a 15 +/- 6-fold increase. In the same RNA samples, GM-CSF increased IL-1 beta mRNA 5.9 +/- 1.7-fold, but IL-4 decreased IL-1 beta mRNA to half that of control levels (0.45 +/- 0.17). Thus, a single stimulus (IL-4) decreased the expression of an agonist (IL-1) while it increased the expression of an antagonist (IRAP). When U937 cells were treated with both IL-4 and GM-CSF, the level of IRAP mRNA induced was additive, suggesting that the cytokines acted differently to increase IRAP mRNA levels. The level of IL-1 mRNA in cells treated with both IL-4 and GM-CSF was intermediate. Dexamethasone and cycloheximide inhibited all mRNA increases and did not reverse IL-4-induced decreases in IL-1 mRNA. These studies have identified two cytokines which induce IRAP in the monocytic cells studied, and have partially characterized the differential regulation of IL-1 and its antagonist, IRAP. PMID- 8419186 TI - Split tolerance of Th1 and Th2 cells in tolerance to Theiler's murine encephalomyelitis virus. AB - Theiler's murine encephalomyelitis virus (TMEV) produces a chronic, inflammatory demyelinating disease in susceptible mouse strains that is used as a model for multiple sclerosis. Because disease susceptibility correlates temporally with the development of virus-specific delayed-type hypersensitivity (DTH) responses, we studied methods and mechanisms by which virus-specific DTH could be specifically inhibited. The intravenous injection of UV-inactivated TMEV coupled to syngeneic splenocytes via a carbodiimide linkage (TMEV-SP), prior to immunization, induced a significant degree of tolerance in virus-specific helper (Th) cells as determined by decreased DTH and T cell proliferative responses, and decreased interleukin (IL)-2 and interferon (IFN)-gamma protein and mRNA levels. In contrast to the reduced levels of Th1-specific lymphokine mRNA levels, IL-4 specific mRNA levels in response to virus stimulation were not affected in tolerant mice. Surprisingly, the total anti-TMEV antibody response in DTH tolerant mice was enhanced 20-100-fold over sham-tolerized controls and was composed of reduced levels of anti-virus IgG2a, but dramatically increased levels of anti-virus IgG1. The "split-tolerance" was antigen specific, dependent on the concentrations of TMEV and carbodiimide used in the coupling procedure, and varied with the number of coupled syngeneic splenocytes administered. The fixative effects of carbodiimide on antigen-presenting function were necessary for the induction of DTH tolerance with TMEV-SP, since intravenous administration of virus coupled to splenocytes via a biotin-avidin linkage led to enhanced virus specific antibody responses, but was unable to inhibit DTH unless concomitantly fixed with carbodiimide. Collectively, the data indicate that Th1 cells (mediating DTH, IL-2 and IFN-gamma production, and helper function for IgG2a production) were specifically anergized, with concomitant stimulation of Th2 cells (producing IL-4 and mediating helper function for IgG1 antibody production). PMID- 8419187 TI - T helper type 2-like cells and therapeutic effects of interferon-gamma in combined immunodeficiency with hypereosinophilia (Omenn's syndrome). AB - We characterized the defects of CD4+ cells in a 17-month-old girl suffering from combined immunodeficiency with hypereosinophilia (Omenn's syndrome). Because the vast majority of peripheral blood CD4+ cells expressed the CD45R0 isoform, we purified circulating CD4+ CD45R0+ cells from the patient and healthy individuals in order to compare their production of cytokines. The patient's CD4+ CD45R0+ cells spontaneously produced high levels of interleukin-5 (IL-5) in vitro (1600 pg/ml after 24 h of culture) and this was associated with the presence of IL-5 in serum (323 pg/ml). After stimulation with phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187, they produced higher levels of IL-4 (306 vs. 55 +/- 4 pg/ml) and IL-5 (2900 vs. 213 +/- 72 pg/ml) and lower levels of IL-2 (17 vs. 63 +/- 17 IU/ml) and interferon-gamma (IFN-gamma) (16 vs. 299 +/- 70 IU/ml) than controls CD4+ CD45R0+ cells. This T helper type 2 (Th2) pattern was confirmed by the detection using reverse polymerase chain reaction of IL-4, IL-5 and IL-10 mRNA within peripheral blood mononuclear cells. During a therapeutic trial with human IFN-gamma (40 micrograms/day) which ameliorated the clinical status of the patient, we observed a down-regulation of the in vivo expression of IL-5 and IL 10, a normalization of the eosinophil count and an improvement of the T cell response to phytohemagglutinin. This observation indicates for the first time that Th2-like cells might be involved in certain forms of congenital immunodeficiency and that IFN-gamma might down-regulate their activities in vivo. PMID- 8419188 TI - Localization of self antigen: implications for antigen presentation and induction of tolerance. AB - The fifth component of complement (C5) is a self antigen expressed in serum of normal mice at a concentration of about 50 micrograms/ml. We have previously shown that C5 is constitutively processed and presented by antigen-presenting cells (APC) in normal mice to induce and maintain complete tolerance in major histocompatibility complex (MHC) class II-restricted T cells. This report addresses the question of whether C5 presentation involves exogenous antigen which has been internalized for processing or whether intracellular, biosynthesized C5 is being presented with MHC class II. Macrophages were found to synthesize, but not secrete C5 in bone marrow chimeras made from irradiated C5 deficient [C5(-)] hosts reconstituted with C5-sufficient [C5(+)] bone marrow [C5(+)-->C5(-)]. In these mice, macrophages are the only source of C5. [C5(+)- >C5(-)] chimeras are not tolerant of C5 and generate C5-specific T and B cell responses upon immunization indistinguishable from those of C5(-) mice. Macrophages from [C5(+)-->C5(-)] chimeras are unable to activate C5-specific T cell hybrids in vitro unlike macrophages from a C5(-) strain that has matured in a C5-expressing environment [C5(-)-->C5(+) chimeras]. This shows that under physiological conditions in vivo intracellular C5 does not get access to the class II presentation pathway and thus, does not induce tolerance in class II restricted T cells. PMID- 8419189 TI - CD3 antigen-mediated calcium signals and protein kinase C activation are higher in CD45R0+ than in CD45RA+ human T lymphocyte subsets. AB - T lymphocytes may be separated into subsets according to their expression of CD45 isoforms. The CD45R0+ T cell subset has been reported to proliferate in response to recall antigen and to mitogenic mAb to a much greater extent than the CD45RA+ subset. This difference could be due to more efficient coupling of the T cell antigen receptor complex to mitogenic signaling pathways. To investigate this possibility, CD3 antigen-induced calcium signals, diacylglycerol (DAG) production and protein kinase C (PKC) activation levels were compared in CD45RA+ and CD45R0+ human T lymphocyte subsets derived from peripheral blood. The mean CD3-induced rise in intracellular calcium was 80% greater in CD45R0+ than in CD45RA+ cells. Basal DAG levels in CD45R0+ cells were found to be, on average, 60% higher than in CD45RA+ cells (p = 0.002), but the CD3-induced production of DAG over background was not different in the two subsets (p = 0.4). Basal PKC activity, and CD3-induced PKC activation levels over background, were found to be 50% and 140% higher, respectively, in CD45R0+ cells than in CD45RA+ cells (p = 0.015 and 0.023). The CD45R0+ subset contained a higher proportion of cells expressing activation markers, such as CD25, CD71 and major histocompatibility complex class II, when compared to the CD45RA+ subset. Our results suggest that the elevated basal DAG levels observed in the CD45R0+ subset may reflect the recent activation of these cells. Both the higher basal DAG and CD3-induced elevation in intracellular calcium observed in the CD45R0+ cells may contribute to the greater PKC activation signals triggered by CD3 mAb in this subset. These findings elucidate the greater response of CD45R0+ T cells to mitogenic stimuli compared to CD45RA+ cells. PMID- 8419190 TI - Comparison of three different forms of HLA-DR4Dw4 proteins. AB - The biochemical behavior and peptide binding properties of a soluble form of the human class II DR4Dw4 molecule (PI-DR4Dw4) were compared to DR4Dw4 molecules containing the transmembrane and cytoplasmic domains that were purified both from B and transfected chinese hamster ovary cells. Recombinant and B cell-derived DR4Dw4 molecules bound monoclonal anti-DR4Dw4 antibodies with different affinities and varied in their stability in the presence of sodium dodecyl sulfate. The three forms of DR4Dw4 bound peptides with a similar apparent affinity constant, but soluble class II molecules bound up to ten times more peptide than DR4Dw4 containing a transmembrane region. Peptide binding kinetics for soluble DR4Dw4 molecules were 10-20 times faster than for the other two forms of DR4Dw4 molecules. Finally, soluble PI-DR4Dw4/peptide complexes were shown to stimulate T cell proliferation. PMID- 8419191 TI - Antigen and helper T lymphocytes activate B lymphocytes by distinct signaling pathways. AB - Resting murine B lymphocytes can be induced to proliferate by cross-linking membrane immunoglobulin, the antigen receptor, or by contact with activated helper T lymphocytes in the absence of a signal through membrane immunoglobulin. Little is known about the molecular nature of contact-dependent T cell help. To determine whether helper T cells activate B cells through different signal transduction and second messenger pathways from those used by membrane immunoglobulin, the effects of drugs which block activation of B cells through membrane immunoglobulin were measured on B cell activation by contact with anti CD3-activated and fixed T helper cells. Cyclosporin A, phorbol esters added at the time of activation, and cAMP agonists all block activation of B cells through membrane immunoglobulin at concentrations at least 100-fold lower than those necessary to block B cell activation by contact with activated Th1 or Th2 helper T cells. Depletion of protein kinase C by pretreatment of B cells with phorbol ester inhibits the proliferative response to anti-immunoglobulin but not the response to contact with activated T cells. The B cell response to lipopolysaccharide is intermediate in sensitivity to cyclosporin A and cAMP agonists, and resembles the response to activated T cells in resistance to phorbol esters and protein kinase C depletion. Various protein kinase inhibitors did not distinguish among these B cell activation pathways, except for the tyrosine kinase inhibitor, herbimycin A, which inhibited anti-immunoglobulin responses at 3- to 5-fold lower concentrations. PMID- 8419192 TI - H-2 I-E molecules isolated from Mls1a stimulatory cells do not activate Mls1a responsive T cells but do present exogenous staphylococcal enterotoxins. AB - The T cell response to allogeneic murine Mls determinants is not H-2 restricted but is dependent on H-2 class II molecules on the Mls-expressing stimulator cells. We have tested planar membranes containing H-2 class II I-E molecules alone or with I-A molecules for their ability to activate a panel of Mls1a specific T hybrids. Despite the ability of the planar membranes to activate an alloreactive T hybrid and to present staphylococcal enterotoxins or an antigenic peptide to appropriately responsive T hybrids, they failed to stimulate the Mls1a specific T hybrids. These findings, in the light of the various controls demonstrating sufficiency of the I-E molecules in the planar membranes, indicate that Mls1a determinants are not covalently bound to I-E molecules; the two molecular species are thus either not physically associated or are linked by a relatively weak interaction. In addition, our experiments show that isolated I-E molecules but not I-A molecules present staphylococcal enterotoxins A and B to two independently derived T hybrids expressing T cell receptor V beta 1, V beta 2 and V beta 6 elements. PMID- 8419193 TI - Bursa-dependent subpopulations of peripheral B lymphocytes in chicken blood. AB - By selective labeling of juvenile chicken bursal cells with colloidal fluorescein isothiocyanate in situ, the emigration rate of bursal lymphocytes to the periphery was estimated at approximately 0.84% and 0.96% of the peripheral blood lymphocyte (PBL) and splenic B cell pool per hour, respectively. Emigrant bursal cells were found primarily in blood and spleen, with very small numbers migrating to thymus, bone marrow, and gut-associated lymphoid tissues. Emigrant bursal cells expressed high levels of both major histocompatibility complex class II antigen and the Ov alloantigen, a phenotype found on a population comprising approximately 4% of bursal cells from which the bursal emigrants may be derived. Surgical bursectomy at 3 weeks of age revealed that peripheral blood B cells could be divided into three distinct populations. Specifically, 60% of the peripheral blood B cells were short lived with a half-life of about 30 h in the blood. These cells accounted for the great majority of emigrants from the bursa to the peripheral blood. Approximately 35% of PBL B cells had a half-life of 12 days following bursectomy and comprised cells which did not divide in the periphery. Consequently, we propose that physiological differences between this population and the majority of bursal emigrants are established intrabursally. The remaining PBL B cells, whose relative proportion increases with age from about 5% of PBL B cells at 2-3 weeks of age, are short lived and are being continually produced from (a) post-bursal site(s) of B cell production. PMID- 8419194 TI - Female reproductive aging--ovarian and uterine factors. AB - OBJECTIVES: To review published data on ovarian and uterine aging in the human female. DATA IDENTIFICATION: Literature review. STUDY SELECTION: Studies examining the effect of age on oocyte quality and uterine receptivity. RESULTS: There is clear evidence in women having in vitro fertilization (IVF) and in normal oocyte donors that oocyte quality declines with age. Three studies of oocyte donation, one study of IVF, and multiple studies in subhuman mammalian species have all shown reduced implantation with aging, and endometrial biopsies in women over age 35 having IVF have shown a high incidence of delayed or absent secretory maturation. In one study of oocyte donation, the reduced rate of pregnancy in women over age 40 was corrected by doubling the dose of progesterone (P) administered to prepare the endometrium for implantation. CONCLUSIONS: Conflicting opinions regarding the effect of aging on uterine receptivity in recipients of donated oocytes appear to have been because of use of high doses of P replacement. When doses more closely approximating physiological levels have been used, a decline of implantation with increasing age has been clearly demonstrated. Oocyte donation, through the use of oocytes from young normal women and an increased level of P replacement to aging recipients, corrects both of these defects. PMID- 8419195 TI - Fertility prognosis for infertile couples. AB - OBJECTIVE: To develop a fertility prognosis model for infertile couples. DESIGN: Prospective follow-up study. PARTICIPANTS: In the period November 30, 1977 to June 1, 1985, 321 consecutive couples were investigated for infertility at Hvidovre University Hospital. Investigation of the female included detection of ovarian, cervical, and anatomic disorders, whereas in the male semen analysis and sperm penetration test (P-test) were performed. Altogether, 108 couples (34%) conceived, whereas 213 (66%) were observed for an average of 2.3 years without having achieved pregnancy. MAIN OUTCOME MEASURE: The Cox regression model was used to predict the time required to conceive based on informations provided by fertility investigations. RESULTS: Three of 16 prognostic variables (the period of infertility, the female infertility factor, and the P-test) possess significant prognostic information. The period of infertility and the P-test are best scored as continuous variables, whereas the female infertility factor are best categorized in four classes, i.e., normal, ovulation or cervical disorder, anatomic disorder, or a combination of disorders. CONCLUSIONS: The three prognostic variables are combined to form a prognostic index that predicts the fecundability of the individual infertile couple. PMID- 8419196 TI - Single-dose pharmacokinetics and pharmacodynamics of recombinant human follicle stimulating hormone (Org 32489*) in gonadotropin-deficient volunteers. AB - OBJECTIVE: To assess safety, pharmacokinetic, and pharmacodynamic properties of recombinant human follicle-stimulating hormone (FSH; Org 32489, Organon International, Oss, The Netherlands) after a single intramuscular injection in the buttock. DESIGN: In a prospective study, safety variables, serum FSH, luteinizing hormone, inhibin, estradiol (females only), and testosterone (males only) were evaluated up to a maximum of 11 days after injection of 300 IU recombinant FSH. SETTING: Four specialist Reproductive Endocrinology and Infertility units. VOLUNTEERS: Fifteen men and women exhibiting all pituitary gonadotropin deficiency. RESULTS: A single bolus of 300 IU recombinant FSH was well tolerated, and no drug-related adverse effects were noted. Comparison of before and after treatment safety variables, including serum antirecombinant FSH antibodies, showed no changes of clinical relevance. Analysis of serum FSH levels revealed comparable elimination half-lives of 44 +/- 14 (mean +/- SD) and 32 +/- 12 hours in women and men volunteers, respectively. In contrast, peak FSH concentrations were significantly lower in women than in men volunteers (4.3 +/- 1.7 versus 7.4 +/- 2.8 IU/L), and the time required to reach peak levels of FSH was significantly longer in women than in men (27 +/- 5 versus 14 +/- 8 hours). The area under the serum level versus time curve tended to be smaller in women than in men volunteers (339 +/- 105 versus 452 +/- 183 IU/L x hours), but the difference did not reach statistical significance. Together these data suggest that recombinant FSH is absorbed from its intramuscular depot to a lower rate and extent in women than in men. In both sexes a relationship between serum FSH levels and body weight was apparent. During the experimental period, other hormones remained low at baseline levels or were only slightly increased. CONCLUSION: Our findings indicate that recombinant FSH is well tolerated and that it is absorbed from its intramuscular depot to a higher rate and extent in men than in women. After intramuscular administration, its half-life is in good agreement with that previously reported for natural FSH. PMID- 8419197 TI - Ovarian sensitivity to follicle-stimulating hormone during the follicular phase of the human menstrual cycle and in patients with polycystic ovarian syndrome. AB - OBJECTIVE: To investigate the existence of a different sensitivity of ovaries to follicle-stimulating hormone (FSH) during the follicular phase of the human menstrual cycle and in patients with polycystic ovarian syndrome (PCOS). DESIGN: Thirty-four normal subjects and 13 patients with PCOS were treated intravenously by FSH (75 or 225 IU) or saline at different stages of follicular phase. MAIN OUTCOME MEASURES: Plasma levels of luteinizing hormone (LH), FSH, estradiol (E2), and testosterone (T) in samples collected for a period of 26 hours after the injection. RESULTS: In patients at the early stages of follicular phase (baseline E2 < 50 pg/mL), FSH increased in dose-dependent manner E2 and E2:T-stimulated area under curve (AUC) in respect to saline experiments. In PCOS subjects, saline E2, and E2:T-stimulated AUC were significantly lower than normal women. Follicle stimulating hormone (75 IU) dramatically increased these values, and no difference was seen in respect to 75 and 225 IU FSH-treated controls. In patients with E2 baseline plasma levels > 50 pg/mL, FSH (75 or 225 IU) failed to increase both E2 and E2:T-stimulated AUC in comparison with saline studies. CONCLUSIONS: Early stages of follicular phase in normal and polycystic ovaries are the most responsive to the elevation of circulating FSH levels, whereas the ovarian sensitivity spontaneously decreases as follicular maturation enhances. PMID- 8419198 TI - Comparison of the methods of artificial insemination on the incidence of conception in single unmarried women. AB - OBJECTIVE: To compare pregnancy rates after intrauterine insemination (IUI) versus pericervical insemination in absolute male factor infertility using each patient as her own control. DESIGN: Ovulatory women with patent fallopian tubes without male partners were alternately inseminated with cryopreserved donor semen using either IUI or pericervical insemination techniques. A total of 81 cycles, which included up to 4 cycles per patient were performed. In this manner a comparison between the efficacy of each method could be evaluated. SETTING: The donor insemination program at the Center For Assisted Reproduction at Northwestern University Medical School. PATIENTS: Twenty-six single, healthy, unmarried women with patent fallopian tubes and < 40 years of age without male partners (absolute male factor infertility). MAIN OUTCOME MEASURES: Positive quantitative serum subunit of human chorionic gonadotropin followed by the presence of an intrauterine gestational sac seen by transvaginal ultrasonography. RESULTS: Fourteen (54%) of 26 patients conceived including two (14%) miscarriages within four insemination cycles. Seven (17.5%) patients after IUI, and 7 (17.1%) patients after pericervical insemination conceived. The pregnancy rates were similar regardless of the order of insemination method. CONCLUSION: These findings reveal that there is no statistical difference in the pregnancy outcome between these two methods of insemination in absolute male factor infertility. PMID- 8419199 TI - The effects of clomiphene citrate and cyclofenil on cervical mucus volume and receptivity over the periovulatory period. AB - OBJECTIVE: To determine the effects of clomiphene citrate (CC) and cyclofenil on cervical mucus (CM) volume and receptivity sampled serially over the periovulatory period. DESIGN: Using prospective luteinizing hormone (LH) timing CM volume and receptivity were compared in standard CC and cyclofenil-stimulated cycles using normal ovulatory cycles as controls. LOCATION: The Donor Insemination Unit at the University Research Clinic, Sheffield, United Kingdom. PATIENTS: Twenty anovulatory patients and 10 normally ovulating patients, all of whom were participating in a treatment cycle of donor insemination. INTERVENTIONS: The 20 anovulatory patients were allocated at random into two groups: group 1 was administered 50 mg of CC on days 2 to 6 of the menstrual cycle; group 2 was administered 400 mg of cyclofenil on days 3 to 12 of the menstrual cycle. All the patients were given a single treatment of donor insemination 24 to 36 hours after the onset of the LH surge. RESULTS: Clomiphene citrate and cyclofenil were shown to exert differential impacts on CM quantity and quality. In terms of quantity, the CC patients produced significantly lower volumes of CM than the cyclofenil patients and controls. In terms of quality, the CC patients and controls produced CM of similar receptivity, whereas the cyclofenil patients produced CM that was significantly more receptive to sperm than both the CC patients and controls. CONCLUSIONS: Neither CC nor cyclofenil exerted a detrimental impact on CM quality throughout the periovulatory period. PMID- 8419200 TI - Inhibin and relaxin concentrations in early singleton, multiple, and failing pregnancy: relationship to gonadotropin and steroid profiles. AB - OBJECTIVE: To examine the inter-relationships between inhibin, relaxin, steroid concentrations, estradiol (E2), progesterone (P), and gonadotropins in early pregnancy. DESIGN: Hormone concentrations in plasma were measured during the luteal phase of subjects who became pregnant (n = 58) or failed to become pregnant (n = 47) after ovarian hyperstimulation for in vitro fertilization gamete intrafallopian transfer (IVF-GIFT) (group 1). A further group of subjects became pregnant (n = 7) or failed to become pregnant (n = 8) during endocrinology tracking of a natural cycle (group 2). Blood was obtained every 3 days in the luteal phase from day 5 in group I (day 0 was oocyte recovery) and from day 0 (first increase in luteinizing hormone [LH]) in group II. RESULTS: Progesterone and E2 were increased over nonpregnant values by day 11 (P) and day 16 (E2) in group I and by day 11 (E2 and P) in group II. Inhibin and relaxin concentrations were significantly increased by day 16 in group I (often by day 11) and by day 14 in group II pregnancy subjects. A direct relationship existed between inhibin, P, relaxin, and human chorionic gonadotropin (hCG). Subjects who had twin pregnancies demonstrated higher concentrations of all hormones and often exhibited increases earlier (by day 11 in group I) than singleton pregnancy subjects. Pregnancies that ended in miscarriages tended to have lower concentrations of P and inhibin. None of the hormones reliably discriminated between the clinical conditions of blighted ovum and of spontaneous abortion, and the predictive value of any hormone measured for miscarriage was not high. CONCLUSIONS: The trend of inhibin and relaxin concentrations closely parallels rises in P during early pregnancy. Luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels are suppressed very early in pregnancy. The suppression of LH and FSH in hyperstimulated cycles is more governed by E2 than inhibin in stimulated cycles. Some subjects destined to miscarry exhibit abnormal endocrine changes very early in the luteal phase. PMID- 8419201 TI - Coculture of mouse embryos with cells isolated from the human ovarian follicle, oviduct, and uterine endometrium. AB - OBJECTIVE: To examine the specificity of somatic cell support by comparing embryonic development during long-term in vitro coculture with feeder cells derived from the human ovarian follicle, oviduct, and endometrium. DESIGN: Comparative study of murine embryo development and degeneration during 6 days of in vitro coculture. RESULTS: All feeder-cell cultures were beneficial to embryonic development and viability. Few differences were observed between feeder cell types (epithelial or fibroblastic) or cell origin (ovarian follicle, oviductal, or endometrial). Embryos developed to the eight-cell stage in 24 hours whether in coculture (83.6% to 100%) or in media alone (85.2%); however, further development in media alone decreased compared with coculture (15.6% versus 63.4% to 87.7%, plating) and embryo degeneration increased (67.9% versus 5.5% to 19.4%) after 6 days. CONCLUSIONS: [1] Coculture of embryos with human reproductive tract cells is beneficial to embryonic development and viability. [2] Human somatic cell support of murine embryos during long-term in vitro coculture is not tissue specific nor dependent on cell type. PMID- 8419202 TI - Infertility: a non-event transition. AB - OBJECTIVE: To determine the predictive validity of four transition variables and seven individual variables in differentiating between individual responses to infertility. DESIGN: Forty-three primary infertile women and 28 men attending a general infertility treatment clinic voluntarily agreed to participate in this cross-sectional study and completed five standardized instruments on a one-time only basis. RESULTS: Adjustment to infertility was related to positive self esteem, internal locus of control, higher socioeconomic status, and moderate age. High levels of anxiety and distress were related to low self-esteem, undifferentiated sex role identity, and advanced age. Actual and perceived duration of infertility were factors also related to participants' responses to infertility. CONCLUSION: Results highlight the importance of psychological assessment and continuing support in the management and treatment of infertility, with concerns being raised regarding the advisability of protracted medical intervention. PMID- 8419203 TI - Subzonal multiple sperm injection in the treatment of previous failed human in vitro fertilization. AB - OBJECTIVE: To assess the value of subzonal sperm injection in previous failed spontaneous in vitro fertilization (IVF). DESIGN: Prospective study on patients with at least one cycle of previous failed spontaneous IVF. SETTING: Human Reproductive Biology Unit, Soliman Fakeeh Hospital, Jeddah, Saudi Arabia. PATIENTS: Thirty-nine patients with prolonged infertility with at least one previous cycle of failure of spontaneous fertilization of any oocyte. RESULTS: Seventy-eight of 276 oocytes successfully injected with three to four sperms were fertilized. Fertilized rate was 28.2%. Polyspermy was observed in 6 (8%) of the fertilized oocytes. Sixty-six (92%) of the 72 with monospermic fertilization cleaved normally and were replaced in the uterine cavity. Pregnancy occurred in 6 (23%) of the women who had embryos replaced. CONCLUSIONS: Multiple sperm injection into the oocyte subzonal space should be considered for the treatment of patients with previous failed spontaneous IVF. PMID- 8419204 TI - Changes in follicular fluid gas and pH during carbon dioxide pneumoperitoneum for laparoscopic aspiration and their effect on human oocyte fertilizability. AB - OBJECTIVE: To determine changes in follicular fluid (FF) gases and acidity (pH) during either ultrasound (US)-guided or laparoscopic aspiration under carbon dioxide (CO2) pneumoperitoneum and relate the findings to the outcome of oocyte fertilization in vitro. DESIGN: A prospective study carried out over a limited period of 1 month during which test FF samples were obtained without direct exposure to CO2 or air. SETTING: Within the assisted pregnancy program setting at the Human Reproductive Biology Unit at the Soliman Fakeeh Hospital. PATIENTS: Infertile women undergoing either laparoscopic or US guided follicular aspiration for the purpose of in vitro fertilization treatment. INTERVENTION: Laparoscopic follicular aspiration under 100% CO2 pneumoperitoneum or transurethral US-guided follicular aspiration. RESULTS: Mean FF carbon dioxide partial pressure (pCO2) and oxygen partial pressure (pO2) were significantly higher in those aspirated at laparoscopy compared with those obtained by US-directed aspiration. The mean FF pH in the laparoscopic group was as a consequence significantly lower than in the US group. Oocyte fertilizability was significantly reduced in the laparoscopic aspiration group compared with the US group. This was more pronounced when the fertilization rate of the last recovered oocytes from the groups were compared. No difference was observed in the post fertilization embryonic development between the two groups. CONCLUSION: One hundred percent pneumoperitoneum leads to changes in FF that adversely affect oocyte fertilizability in a time-dependent manner. The use of US-directed procedure is advocated for all patients undergoing follicular aspiration in assisted pregnancy program. Where this is not feasible, the duration of CO2 pneumoperitoneum should be minimized. PMID- 8419205 TI - No detrimental effects in delaying initiation of gonadotropin administration after pituitary desensitization with gonadotropin-releasing hormone agonist. AB - OBJECTIVE: To determine if delaying initiation of exogenous gonadotropin administration after pituitary desensitization with gonadotropin-releasing hormone agonist (GnRH-a) is a realistic option to avoid scheduling clinical and laboratory work on weekends/holidays. DESIGN, PATIENTS: A review of 57 in vitro fertilization (IVF) cycles in which, after pituitary desensitization with GnRH-a, initiation of gonadotropin administration were delayed in an attempt to avoid off hour work. Thirty-eight IVF cohort cycles served as control. SETTING: Tertiary medical center. RESULTS: There were no statistically significant differences in ovarian response, dose of gonadotropin required, oocytes and embryos obtained, pregnancy rates, and abortion rates between groups. Eighty-three percent of the delayed cycles had clinical and laboratory work that fell within weekdays. CONCLUSION: Delaying initiation of exogenous gonadotropin administration after pituitary desensitization had no detrimental effects on IVF outcomes. It may be used to avoid scheduling work on weekends/holidays. PMID- 8419206 TI - Placental protein 14 secretion during in vitro fertilization cycles with and without human chorionic gonadotropin for luteal support. AB - OBJECTIVE: To investigate levels of placental protein 14 (PP14) in in vitro fertilization (IVF) patients with and without exogenous human chorionic gonadotropin (hCG) for luteal support. DESIGN, PATIENTS: Thirty-one women undergoing IVF were studied. For 18 women, hCG was administered in the luteal phase, and 12 became pregnant. Five pregnancies occurred in 13 women not receiving exogenous hCG. SETTING: All the patients attended the University of Southampton/Chalybeate Hospital IVF program. RESULTS: There was no change in PP14 levels 2 days after embryo transfer (ET), but small significant rises were noted by day 8 in all patients. Thereafter, levels rose further in pregnant subjects but showed no change in nonpregnant patients. The highest level of PP14 was seen in the group of women on hCG support, but there was no overall statistical difference between those on support and those not. In the nonpregnant group, there was no significant correlation between progesterone (P) and PP14 8 days from ET, whereas a highly significant correlation was noted in the pregnant group. CONCLUSIONS: Neither hCG nor P are primary factors in the control of endometrial PP14 secretion, but PP14 and P may have common underlying control mechanisms. PMID- 8419207 TI - The mouse embryo culture system: improving the sensitivity for use as a quality control assay for human in vitro fertilization. AB - OBJECTIVE: To determine whether the mouse embryo culture system can be sensitized to provide improved differentiation of suboptimal culture media for in vitro fertilization. DESIGN: Mouse embryo development in media prepared from one of three water sources were compared using embryos from two mouse strains, culturing embryos from either zygote or two-cell stage, and pretreating with either zona removal and/or cryopreservation. SETTING: Academic research department, tertiary care referral center. RESULTS: Embryos from CD1 mice were able to develop in suboptimal culture conditions, even when pretreated with zona removal or cryopreservation. Embryos from B6CBA/F1J mice were more sensitive to suboptimal culture conditions when harvested at the zygote stage than at the two-cell stage, and this sensitivity was improved after zona removal before culture. CONCLUSIONS: The mouse embryo culture system has deficiencies as an assay of culture medium quality, but the sensitivity of the assay can be optimized by harvesting at the zygote stage from an appropriate strain and by zona pellucida removal before culture. PMID- 8419208 TI - The role of aerobic and anaerobic semen cultures in asymptomatic couples undergoing in vitro fertilization: effects on fertilization and pregnancy rates. AB - OBJECTIVE: To determine if routine semen culture is useful in asymptomatic couples undergoing in vitro fertilization and embryo transfer (IVF-ET). DESIGN: Prospective data collection. SETTING: All cultures and IVF cycles were performed at the University of North Carolina in Chapel Hill, North Carolina. PARTICIPANTS: All asymptomatic couples undergoing IVF-ET from January 1989 through January 1990. INTERVENTIONS: Aerobic and anaerobic cultures were performed on semen samples obtained before IVF. MAIN OUTCOME MEASURES: Quantitative semen cultures were evaluated for both aerobic and anaerobic bacterial isolates. Fertilization and pregnancy rates (PRs) were compared in patients with positive and negative semen cultures. RESULTS: Eighty percent of cultures contained at least one bacterial isolate. Three of the four most commonly isolated bacteria were normal skin flora. Positive culture results had no effect on either fertilization or PRs. CONCLUSIONS: Bacterial contamination is common with semen collection, yet routine semen cultures are not beneficial in asymptomatic couples undergoing IVF ET. PMID- 8419209 TI - A new computerized method of reading sperm morphology (strict criteria) is as efficient as technician reading. AB - OBJECTIVES: To compare the ability of a computerized method of sperm morphology with the manually recorded method in predicting in vitro fertilization (IVF) results, to compare results obtained by both methods, and to determine the intraobservation variability. DESIGN, SETTING, PATIENTS: Forty-three stained semen slide preparations from two large level-three academic institutions' reproductive endocrinology units (IVF programs) were blindly evaluated, and the sperm were classified into normal and amorphous forms. RESULTS: Experiment 1: Twenty-one slide preparations from the Tygerberg gamete intrafallopian transfer program were manually evaluated; the fertilization rates for the groups with < 14% and > 14% normal sperm forms were 33.3% (15/45 oocytes) and 76.6% (46/60 oocytes), respectively. Corresponding fertilization rates with FERTECH were 46.8% (30/64) and 75.6% (31/41). Experiment 2: Twenty-two slide preparations from the Norfolk IVF program were evaluated. The manual method reported a fertilization rate in the group with < 14% normal forms of 27.4% (14/51 oocytes) compared with 90.0% (127/141 oocytes) in the group with > 14% normal forms. Corresponding figures for the FERTECH method were 33.9% (18/53) and 88.4% (123/139), respectively. Experiment 3: When the 43 slide preparations were blindly evaluated using both methods, 84% of the FERTECH evaluations correlated well with the manual method and FERTECH ability to diagnose the subfertile male (< 14% normal forms) was 95% (sensitivity). Experiment 4: A total of 16 different slides (8 per group) were randomly selected and analyzed five times (100 cells per reading) by the computerized method. The slides were obtained from men with normal sperm morphology of < 14% and > 14% as classified by the manual method. In the first group (< 14%) 97.5% (39/40) of the readings classified the sperm in the proper category, whereas in the second group (> 14%) 95% (38/40) of the cases were correctly identified. CONCLUSION: Using strict criteria for morphology evaluation, there is a positive and significant correlation between FERTECH evaluation and manual assessment. The reproducibility of the computerized method and the ability to distinguish between fertile and subfertile groups using those criteria are good. PMID- 8419210 TI - Pentoxifylline is not useful in enhancing sperm function in cases with previous in vitro fertilization failure. AB - OBJECTIVE: To assess the efficiency of the addition of pentoxifylline to spermatozoa for in vitro fertilization (IVF) in cases with previous fertilization failure. DESIGN: After retrieval, oocytes were divided at random and inseminated with spermatozoa prepared with or without pentoxifylline. SETTING: In vitro fertilization program of the Centre for Reproductive Medicine of the Dutch speaking Brussels Free University, Belgium, a tertiary referral institution. PATIENTS: Couples in this study were undergoing a second or third IVF attempt for the treatment of male factor infertility. INTERVENTIONS: Sperm samples were divided into two equal parts. The first part was prepared with 3.6 mM pentoxifylline (treatment group) and the other part without pentoxifylline (control group). After swim-up, oocytes were randomly inseminated with spermatozoa from either treatment. MAIN OUTCOME MEASURES: Fertilization and cleavage rates after IVF. RESULTS: Overall a fertilization rate of 8.8% was achieved with no significant differences between treatment and control groups. Cleavage rate was 81.5% and was not different between groups. CONCLUSIONS: This study demonstrates no therapeutic advantage of using pentoxifylline in IVF for male factor infertility in cases with previous fertilization failure. PMID- 8419211 TI - The use of clomiphene citrate in the treatment of azoospermia secondary to incomplete androgen resistance. AB - An infertile male with a deletion within the AR gene is discussed. The patient presented with azoospermia and, after daily CC treatment, was found to have sperm within his ejaculate. However, the ultimate goal of pregnancy was not achieved, nor were there enough sperm present to warrant an IVF attempt. PMID- 8419212 TI - Vaginosonography for recording of cycle-related myometrial contractions. AB - It has been recognized from experimental or invasive studies that the nonpregnant human uterus has an inherent contractibility. We used vaginosonography for imaging contractions of the inner third of the myometrium. The direction, frequency, and symmetry of contractions were noted. We studied 53 women and subdivided them into four groups based on the cycle phase. During menstruation we found contractions toward the cervix with irregular frequency varying between 1 and 3/min. In the periovulatory period we noted the highest frequency of 10/min of regular contractions toward the fundus. The results showed that active myometrial contractions can be detected sonographically throughout the whole menstrual cycle. Increased myometrial contractions toward the fundus in the periovulatory period may be involved in sperm transport to the tubes. PMID- 8419213 TI - Intraocular endometrium in the rabbit as a model for endometriosis. AB - The model described suggests that endometrium can be successfully transplanted to the rabbit eye and observed through a slit lamp for morphological changes such as vascularization. Sampling of aqueous humor in volumes adequate for biochemical measurements have been demonstrated. Autografts of rabbit endometrium survived for up to 181 days. Although xenografts of human endometrial and endometriotic tissue demonstrate some adherence and vascularization, there is indication of immune rejection by day 7. Other treatment regimens will be explored with the objective of prolonging the graft survival time. PMID- 8419214 TI - Ultrasound-guided transcervical tubal catheterization for assisted reproduction: a learning program using laparoscopy for confirmation. AB - The result of this pilot study confirmed that the US-directed transcervical tubal catheterization procedure for assisted reproduction can be learned over a short period of time and may well produce comparable PRs as seen with laparoscopic transfer. However, practice in the technique with confirmation of placement by laparoscopy is advised before incorporating this procedure into a program of assisted reproduction. PMID- 8419215 TI - Sperm antibodies--"gold standard?". PMID- 8419216 TI - Sperm engines--why don't we know more? PMID- 8419217 TI - Critical comparisons of alternative therapies for ectopic pregnancy! PMID- 8419218 TI - Critical comparisons of alternative therapies for ectopic pregnancy! PMID- 8419219 TI - Benefit of leukocyte immunizations? PMID- 8419220 TI - Laparoscopic endometriosis treatment: is it better? AB - OBJECTIVE: To assess the hypothesis that pregnancy rates (PRs) after operative laparoscopy (Laparoscopy Group) for endometriosis treatment would be equal to or greater than diagnostic laparoscopy only (No Treatment Group), diagnostic laparoscopy with medical treatment (Medical Treatment Group), and laparotomy (Laparotomy Group). DESIGN: Prospectively recorded data were analyzed to identify significant variables affecting PRs. These variables were statistically controlled for using survival analysis with multiple fixed covariates to compare operative laparoscopy PRs versus other treatment PRs. SETTING: Treatment was performed by the senior author in a referral reproductive endocrinology and surgery private practice. PATIENTS: Five hundred seventy-nine infertile women were diagnosed with endometriosis. A subset (n = 258) considered to have endometriosis only was evaluated separately (Endometriosis-Only Subset). INTERVENTIONS: Treatment groups included: No Treatment Group, Medical Treatment Group, Laparoscopy Group, and Laparotomy Group. MAIN OUTCOME MEASURE(S): Pregnancy was used as the indicator of treatment success. RESULTS: Laparoscopy Group PRs were at least equal to all other treatment groups and were significantly higher than some other treatment groups in some comparisons. CONCLUSIONS: Operative laparoscopy is the treatment of choice for infertile women with endometriosis unless they have severe tubal and/or fimbrial disease. PMID- 8419221 TI - Transvaginal-transmyometrial embryo transfer: the Towako method; experiences of 104 cases. AB - A new technique of ET based on US-guided transmyometrial puncture has been performed in 104 cases. Thirty-eight patients conceived for a clinical PR of 36.5% per attempt. The use of this technique is proposed to overcome problems of difficult transfers because of cervical abnormalities. No serious complications were observed. PMID- 8419222 TI - Is there a place for the crossover design in infertility trials? PMID- 8419223 TI - Differential distribution of glycoconjugates in human reproductive tract. AB - OBJECTIVES: To study the distribution of glycoconjugates in the epithelia of various regions of the human female reproductive tract and to examine whether lectins can be used as specific probes to define cell populations in the human female reproductive tract. DESIGN: Nineteen fluorescein isothiocyanate (FITC) labeled lectins, which recognize different oligosaccharide structures, were applied to frozen sections of human fallopian tubes (distal and proximal segments) and endometrium. The differential binding patterns of the lectins to the epithelia were evaluated under fluorescent microscopy. RESULTS: Four of 19 FITC-labeled lectins tested demonstrated distinct binding patterns along the reproductive tract. Arachis hypogaea (PNA) and Ricincus communis II selectively reacted with epithelial cells of endometrium and distal, but not proximal tube. In addition, PNA distinguished ciliated from secretory cells of the distal tube. Maclura pomifera binding followed a gradient of decreasing intensity from distal to proximal tube and was negative in the endometrium. Dolichos biflorus reacted exclusively with proximal tubal epithelium. CONCLUSIONS: Analysis of the FITC lectin binding pattern and the sugar specificities indicate that the distribution of galactosyl residues varies among different regions of the reproductive tract, whereas glucosyl, mannosyl and fucosyl residues are more evenly distributed. The differential expression of glycoconjugates may be involved in the distinct biological function of each region in female reproductive tract. PMID- 8419224 TI - An analysis of social and psychological characteristics of women volunteering to become oocyte donors. AB - OBJECTIVES: To design a recruitment program and analysis to determine social and psychological characteristics of women who volunteer to be oocyte donors; to evaluate consistency between clinical interviews and psychological testing; and to determine whether socially and psychologically normal women tend to volunteer. PARTICIPANTS: Over a 2-year period 95 women were recruited from a middle-class population surrounding the medical center. Candidates were evaluated on the basis of clinical interviews and performance on the Minnesota Multiphasic Personality Inventory. Seventy-three percent were accepted, and 63% donated oocytes. INTERVENTIONS: Interviews and psychological test evaluation were conducted independently by a clinical sociologist and a clinical psychologist; selections were made in a meeting of the treatment team. RESULTS: Interview and test data demonstrated consistency and indicated that women who volunteer tend to be socially conventional, outgoing, and free from psychopathology. The typical donor tends to be 26 years old, married with one or two children, holds religious or spiritual beliefs, has 2 years of college, works at least part-time in a white collar job, and is a person with high energy who has additional interests. Incidence of dysfunction in family of origin or family of orientation (abandonment, abuse) is unremarkable. Although motherhood is highly valued by oocyte donor volunteers, in test performance this group does not endorse traditional female role stereotypes. CONCLUSIONS: We have determined that this recruitment protocol is reliable for screening normal oocyte donors and would recommend its use in programs involved in oocyte donation. PMID- 8419225 TI - The effect of endometrial thickness and echo pattern on in vitro fertilization outcome in donor oocyte-embryo transfer cycle. AB - There have been some conflicting data concerning the importance of endometrial thickness and echo patterns before transfer in different IVF-ET situations under different COH regimens. We previously found in women undergoing IVF-ET after luteal phase LA-hMG a significantly higher PR in those patients attaining at least a 10-mm endometrial thickness and a lower rate in those women with an entirely homogeneous hyperechogenic endometrium (pattern C). The present study evaluated the relationship of endometrial thickness and echo pattern to PRs in donor oocyte recipient immediately before transfer. There were 16 pregnancies in 58 cycles (27.5%). Conclusions similar to the previous COH study were reached concerning the > or = 10-mm thickness levels correlating with improved PRs (9% versus 38.7%, P < 0.01). In contrast, no correlation with echo pattern was found. PMID- 8419226 TI - Long-term effects of transdermal estradiol with and without medroxyprogesterone acetate. AB - OBJECTIVE: To study the long-term biological and metabolical effects of estradiol (E2) administered by transdermal therapeutic systems with and without the addition of medroxyprogesterone acetate (MPA). DESIGN: Open, randomized, comparative trial. SETTING: The reproductive endocrine unit of a tertiary care university-affiliated hospital. PATIENTS: Fifty-seven postmenopausal women were given E2 transdermally, whereas 28 were randomized to take MPA by mouth. Fifteen premenopausal women were studied for comparison. INTERVENTIONS: Estradiol, 0.1 mg, was administered by a transdermal therapeutic system for 24.5 of 28 days and was cycled for 96 weeks. Medroxyprogesterone acetate, 10 mg, was given for days 13 to 25 of each 28-day cycle (E+P group), whereas the remainder received E2 only. MAIN OUTCOME MEASURES: Serum E2, estrone (E1), luteinizing hormone, follicle-stimulating hormone, low-density, high-density, very low-density, and total cholesterol, triglycerides, blood pressure, renin substrate, plasma renin activity, and serum aldosterone levels were measured in all subjects at baseline and in the postmenopausal women every 24 weeks until the end of study. RESULTS: Mean +/- SE levels of E2 rose significantly from baseline at 24 weeks to 426 and 355 pmol/L for the E only and E+P groups, respectively. Smaller increases of estrone (E1) were observed to 263 and 244 pmol/L for the same respective groups. As expected, baseline levels of both gonadotropins were elevated, fell significantly with E2 administration, but remained increased in comparison with values observed in younger women. Decreases of total and low-density lipoprotein (LDL) cholesterol were observed in both groups that reached statistical significance at 48 weeks or later with the exception of LDL cholesterol in the E only group. No significant change of high-density lipoprotein or very low-density lipoprotein cholesterol or triglycerides was observed. There were reductions of mean systolic and diastolic blood pressures in both groups that reached significance at 72 weeks. Mean baseline plasma renin substrate, plasma renin activity, and serum aldosterone levels were within the ranges observed in younger, healthy women and did not change significantly with E2 administration in either group. CONCLUSION: These data support the long-term efficacy and safety of this form of replacement therapy, particularly in combination with MPA, in women with a uterus. PMID- 8419227 TI - The potential relevance of growth hormone to female reproductive physiology and pathophysiology. AB - OBJECTIVE: To assess possible interfacing between the somatotrophic and reproductive axes. DESIGN: Literature review. MAIN OUTCOME MEASURES: Ovarian growth hormone reception and action. RESULTS: The available literature strongly supports a permissive role for the somatotrophic axis in the reproductive process. CONCLUSIONS: Although a role for growth hormone in reproductive biology appears highly likely, its relevance to the process of puberty and to the normal workings of the menstrual cycle, as well as its possible application in reproductive pathology must await further investigation. PMID- 8419228 TI - Histologic features associated with hormonal responsiveness of ectopic endometrium. AB - OBJECTIVE: To correlate histologic parameters of endometriosis with hormonal responsiveness. DESIGN: Seven hundred sixty-eight unselected endometriotic implants and the corresponding intrauterine endometrium from 196 patients were classified by standard endometrial dating criteria. In addition, other histologic characteristics of endometriotic implants such as the amount of stroma, amount of fibrosis, the presence of surface epithelium, presence of focal hemorrhage, and gland characteristics were also noted. SETTING: Academic tertiary referral center. MAIN OUTCOME MEASURES: Comparison of histologic dating between endometriotic implants and the corresponding endometrium as a function of histologic parameters. RESULTS: Implants that were synchronous with the corresponding eutopic endometrium had more stroma than those that were out of phase. The amount of fibrosis was inversely related to hormonal responsiveness. The presence of surface epithelium in implants was also associated with an impaired response (28.0% versus 48.0% in phase). Endometriomas were found to be in phase with the corresponding endometrium less often than other types of implants (21.7% versus 43.3%). Although endometriomas had similar amounts of stroma when compared with other implants, they had significantly more fibrosis (850.2 microns versus 195.0 microns). CONCLUSIONS: These results suggest that the unpredictable response of endometriotic implants to cyclic endogenous hormones and hormonal therapy may be related to the architectural relationships between the cellular elements found in normal endometrium. PMID- 8419229 TI - In vitro CA-125 secretion by endometrium from women with advanced endometriosis. AB - OBJECTIVE: To determine if the endometrium of women with endometriosis is a potential source of the elevated serum concentrations of CA-125 associated with endometriosis. DESIGN: Secretion of CA-125 by the endometrium in tissue culture was documented and measured in response to treatment with progesterone (P), 17 beta-estradiol (E2), and progesterone and estradiol (E2+P). Sequential biopsies of the endometrium from the early and late luteal phase were obtained from women with laparoscopically documented normal anatomy or moderate to severe endometriosis. SETTING: Division of Reproductive Endocrinology, Department of Obstetrics and Gynecology at the University of Vermont, College of Medicine, Burlington, Vermont. MAIN OUTCOME MEASURES: Secretion of CA-125 by endometrium in vitro was determined by radioimmunoassay in spent media using a commercially available kit (Centicor CA-125 Kit; Centicor, Malvern, PA). RESULTS: Secretion of CA-125 by endometrium in vitro is inhibited by the presence of P and E2+P in both the early and the late luteal phase. The amount of CA-125 secreted increases significantly from the early to the late luteal phase in all treatment groups. When compared with controls, the endometrium of women of with endometriosis secreted two to four times more CA-125 in all treatment groups, in both the early and late luteal phase. CONCLUSIONS: When compared with control endometrium, the endometrium of women with advanced endometriosis secretes significantly more CA 125 and represents a potential source of the elevated serum levels of CA-125 seen in these women. PMID- 8419230 TI - Heparin-sepharose binding growth factors in peritoneal fluid of women with endometriosis. AB - OBJECTIVE: To characterize and to purify the growth substances for mouse embryo fibroblasts (NIH/3T3) and endometrial-like cells of the peritoneal fluid of women with endometriosis. DESIGN: A pool of 102 mL of peritoneal fluid (PF) collected from 24 women with laparoscopic evidence of endometriosis was dialyzed and thereafter chromatographed onto carboxymethyl (CM)-sepharose, heparin-sepharose, cartridges of C18 silica, and reverse-phase high-performance liquid chromatography (reverse-phase HPLC). Mitogenic activity throughout various steps of chromatography was monitored by the increase of the (3H)-thymidine incorporation into NIH/3T3 fibroblasts and enzymatically isolated epithelial cells of rabbit endometrium. RESULTS: Peritoneal fluid of women with endometriosis contained mitogen(s) for NIH/3T3 fibroblasts that were retained on and eluted from CM-sepharose, heparin-sepharose, cartridges of C18 silica, and reverse-phase HPLC columns performed sequentially. Mitogenic material purified 2,500-fold exerted bioactivity on NIH/3T3 and the enzymatically isolated epithelial cells from rabbit endometrium. CONCLUSION: Peritoneal fluid of women with endometriosis contains heparin-binding growth factors that can be purified by sequential chromatography on CM-sepharose, heparin-sepharose, and reverse phase HPLC columns. The presence in the PF of potent mitogens for fibroblasts and endometrium-derived epithelial cells could play an important role in the pathogenesis of endometriosis. PMID- 8419231 TI - Factor XII (Hageman) deficiency in women with habitual abortion: new subpopulation of recurrent aborters? AB - OBJECTIVE: To examine the possible association between factor XII (FXII) deficiency and an elevated number of abortions. DESIGN: Factor XII activity, FXII antigen concentration, other blood clotting parameters, and phospholipid antibodies were examined in venous blood from 43 women with repeated (3 to 7) abortions before the 28th week of gestation but without gynecological and chromosomal abnormalities. The data were compared with those obtained from 49 age matched women without fetal loss. RESULTS: Eight cases with moderately reduced FXII activity (35% to 68% of normal) could be identified in the abortion group, whereas among controls no abnormalities in FXII activity and antigen concentration were found. The relative occurrence of reduced FXII level was higher among patients with more than three abortions as compared with those with three abortions. CONCLUSION: Repeated abortions may be associated with reduced level of FXII activity of unknown origin. PMID- 8419232 TI - Effect of 16,16-dimethyl prostaglandin E2 on lipidic organelles of rat gastric surface mucous cells. AB - BACKGROUND: We examined the effect of 16,16-dimethyl prostaglandin E2 (dmPGE2) on the subcellular distribution of phospholipids in rat gastric surface mucous cells (SMCs) using the cytochemical stain, iodoplatinate (IP). METHODS: The volume of a number of subcellular organelles and the density of IP reactivity within these organelles was determined by modified vertical section stereology. RESULTS: The volume occupied by most of the subcellular organelles was not affected by dmPGE2 treatment, with the exception that the volume fraction of two classes of large infranuclear inclusion bodies (LIIB1 and LIIB2, organelles we have previously shown to contain hydrophobic lipids), were significantly expanded by a factor of 3-4-fold. The distribution of IP reactivity among the various subcellular compartments appeared to undergo a shift in response to dmPGE2 treatment. Once again the major prostanoid effect was seen in the infranuclear inclusion bodies, as the volume density of IP reactivity was increased 2-24-fold in LIIB1 and 9-10 fold in LIIB2 in comparison to control values. CONCLUSION: dmPGE2 administration to rats induces an increase in the volume and IP reactivity of a family of lipid containing organelles, which may underlie its ability to increase the hydrophobic surface properties of the stomach. PMID- 8419233 TI - Mechanism of gastric hyperemia induced by intragastric hypertonic saline in rats. AB - BACKGROUND: Intragastric hypertonic (2 mol/L) saline produces injury in the gastric mucosa and a significant increase in gastric blood flow (hyperemia) in anesthetized rats. We studied the mechanism of this hyperemia. METHODS: Rats were treated with intravenous boluses of NG-nitro-L-arginine methyl ester (3 mg/kg) to block synthesis of endogenous nitric oxide, pyrilamine (1 mg/kg) to inhibit H1 receptors, or indomethacin (5 mg/kg) to block synthesis of endogenous prostaglandins during blood flow studies or with subcutaneous capsaicin (125 mg/kg) 10-14 days before blood flow studies to ablate capsaicin-sensitive afferent nerves. Gastric mucosal blood flow was measured by hydrogen gas clearance before and during intragastric administration of 2 mol/L saline. RESULTS: The gastric hyperemia induced by intragastric 2 mol/L saline was completely blocked only by indomethacin. The associated gastric mucosal damage was increased significantly. CONCLUSIONS: In the rat stomach, the gastric hyperemia induced by intragastric 2 mol/L saline is mediated by endogenous prostaglandins and plays a protective role. Endogenous nitric oxide, H1 receptors, and capsaicin-sensitive afferent nerves are not involved in this protective hyperemia. PMID- 8419234 TI - Trefoil peptide gene expression in gastrointestinal epithelial cells in inflammatory bowel disease. AB - BACKGROUND: This work expands on recent observations that the trefoil peptides pS2 and human spasmolytic polypeptide (hSP) are expressed in the ulceration associated cell lineage (UACL) glands developing in chronic ulcerative conditions. METHODS: Trefoil peptide expression in small intestinal Crohn's disease was examined by in situ hybridization to reveal sites of expression of the messenger RNAs encoding pS2 and hSP and by immunohistochemistry and immunoelectron microscopy to localize the peptides in the UACL and adjacent goblet and neuroendocrine cells. RESULTS: Goblet cells near the UACL expressed pS2 messenger RNA and peptide; ultrastructural immunolocalization revealed pS2 copackaged within mucous cell granules. Neuroendocrine cell hyperplasia was marked in crypts near the UACL; pS2 was copackaged with the neuroendocrine granules. CONCLUSIONS: Copackaging of a secretory protein, pS2, in both mucous and neuroendocrine granules, which have different functions, is unusual and indicates an important role for pS2 in the secretory process itself or as a ligand delivered to its receptors via different routes. It is concluded that trefoil peptides are of considerable potential functional importance in inflammatory bowel disease. PMID- 8419235 TI - Projections of nerve cells from the duodenum to the sphincter of Oddi and gallbladder of the Australian possum. AB - BACKGROUND: This study investigated the existence of direct neural connections between the duodenum and the biliary tract in the Australian possum. METHODS: Retrogradely transported neuronal dyes, Fast Blue and Dil, were injected into the wall of the gallbladder and the sphincter of Oddi. The duodenum, biliary tract, and sympathetic and sensory ganglia were examined for the presence of labeled cell bodies. RESULTS: Two to 3 weeks after gallbladder injection, labeled nerve cell bodies were found in the myenteric plexus of the proximal duodenum but were rare in the duodenum distal to the sphincter of Oddi. No neurons were found in the submucous plexus. Labeled nerve cells were also found in the sphincter of Oddi. After injection of the sphincter, labeled neurons were in both the submucous and myenteric plexuses of the duodenum, on either side. Approximately one third of labeled myenteric neurons were immunoreactive for enkephalin. Labeled cell bodies were also in the coeliaco-mesenteric, nodose, and dorsal root ganglia after both gallbladder and sphincter injection. After a myotomy on the proximal duodenum, no neurons were labeled on the pyloric side of the lesion by subsequent sphincter injection of dye. CONCLUSIONS: Direct neural pathways connect the duodenum with the gallbladder and the sphincter of Oddi, and the sphincter with the gallbladder; this implies that enteric nerve circuits participate in coordinating duodenal and biliary functions. PMID- 8419236 TI - Increased risk of 'high-risk' colorectal adenomas in overweight men. AB - BACKGROUND: Epidemiologic studies have suggested that the incidence of colorectal carcinoma may be related to overnutrition, but retrospective analysis of its relation to the body mass index (BMI: kg/m2) has produced conflicting data. METHODS: To avoid as many sources of statistical bias as possible, the relation between BMI and the presence of colorectal adenomas was investigated in a cross sectional study. RESULTS: Two thousand twelve consecutive colonoscoped patients were investigated (532 patients with malignancies or other conditions associated with weight loss were excluded). The relation between BMI and observed colorectal adenomas was evaluated by a logistic model controlling for other prognostic factors such as age, sex, and serum cholesterol level. The subgroup of "high risk" adenomas with an increased risk of malignant transformation was positively associated with the BMI in men of the age group 50.5-68.1 years (quintiles III and IV: odds ratio for the top quintile vs. the lowest quintile, 3.21; 95% confidence interval, 1.15-8.98). CONCLUSIONS: It was concluded that the risk of developing high-risk adenomas tends to be increased in men who are overweight and that this association is independent of the positive association with the serum cholesterol level recently described. PMID- 8419237 TI - Increased serum deoxycholic acid levels in men with colorectal adenomas. AB - BACKGROUND: Epidemiological and animal studies have suggested that the secondary bile acid deoxycholic acid is cocarcinogenic in colorectal cancer, but this hypothesis was not confirmed by case-control studies investigating fecal bile acids. METHODS: Individual serum bile acid concentrations were investigated in 25 men and 25 women with colorectal adenomas and in an equal number of age- and sex matched controls by gas-liquid chromatography. RESULTS: Deoxycholic acid levels were significantly higher in the sera of men with colorectal adenomas (1.70 +/- 0.59 vs. 1.16 +/- 0.39 mumol/L, P < 0.0005) and in a combined analysis of both sexes (1.47 +/- 0.78 vs. 1.08 +/- 0.39 mumol/L, P < 0.0025). Six- and 12-month follow-up measurements of deoxycholic acid concentrations in a subgroup of 22 men and 17 women showed higher serum levels in men with adenomas, indicating that measurement of deoxycholic acid concentration may be a reliable parameter to investigate its pathogenetic role in colonic neoplasia. CONCLUSIONS: The data of this study support the hypothesis that deoxycholic acid may play a role in the pathogenesis of colorectal cancer. PMID- 8419238 TI - Deglutitive tongue action: volume accommodation and bolus propulsion. AB - BACKGROUND: Swallow function is best analyzed in components because discrete component failure may be compensated for with devised maneuvers, postures, or biofeedback techniques. The present investigation examined normal deglutitive tongue function. METHODS: Biplane videofluoroscopy synchronized with intraluminal manometry was performed in eight volunteers. Tongue surface motion was characterized as centripetal or centrifugal along seven equiangular rays emanating from the tongue center during 1-, 5-, 10-, and 20-mL swallows. RESULTS: The tongue perimeter remained in contact with the alveolar ridge while the central groove exhibited centripetal and subsequent centrifugal motion that, in conjunction with the pharyngeal walls, created an oropharyngeal propulsive chamber and then expelled that chamber's contents into the hypopharynx. Intrabolus propulsive pressure was generated when the initially expansive propulsive chamber volume contracted to the test bolus volume. Because pharyngeal chamber action cycle timing was relatively constant among bolus volumes, vigorous expulsion occurred with large volumes but relatively delayed, sluggish expulsion occurred with smaller volumes. CONCLUSIONS: Deglutitive tongue functions include bolus containment, volume accommodation, and the major contributor to bolus propulsion. PMID- 8419239 TI - Passive jejunal bile salt absorption alters the enterohepatic circulation in immature rats. AB - BACKGROUND: Developmental changes in passive bile salt absorption may alter the enterohepatic circulation. METHODS: 14-, 21-, and 40-day-old anesthetized male Sprague-Dawley rats were studied. Jejunum and ileum were isolated, cannulated, and injected or perfused with a taurocholate, [3H]taurocholate, and nonabsorbable marker solution. Bile was collected. RESULTS: Using bolus injection, jejunal taurocholate absorption rates and total taurocholate absorption were nonsaturable, linearly related to taurocholate dose, and decreased from 14 days (1.62 nmol.cm-1.min-1) to 21 days (1.05 nmol.cm-1.min-1) and 40 days (0.54 nmol.cm-1.min-1). While total taurocholate absorption decreased (14 days, 52.4%; 21 days, 43.7%; 40 days, 30.5%), hepatic taurocholate clearance increased (14 days, 18.2%; 21 days, 23.7%; 40 days, 37.3%). Hepatic taurocholate clearance was saturated only at 14 days. Using jejunal perfusion, total taurocholate absorption (14 days, 62.0%; 21 days, 43.1%; 40 days, 45.3%) and taurocholate absorption rate decreased with age (14 days, 941.13 nmol.cm-2.min-1 per micromole of taurocholate; 21 days, 411.28 nmol.cm-2.min-1 per micromole of taurocholate; 40 days, 334.50 nmol.cm-2.min-1 per micromole of taurocholate). CONCLUSIONS: Passive jejunal bile salt absorption and decreased hepatic bile salt clearance could account for the low intraluminal and high serum bile salt levels observed in immature animals and in human neonates. PMID- 8419240 TI - Epidemiology and prognosis of anorectal melanoma. AB - BACKGROUND: The epidemiology of anorectal melanoma has not been well studied despite its potential relevance to the cutaneous melanoma epidemic, and prognostic studies have generally been based on referred cases. METHODS: Data reported to nine population-based registries in the United States during 1973 through 1987 were analyzed. RESULTS: Fifty-five cases were reported (0.017 x 10( 5)/year). The incidence among blacks was higher but not significantly higher than among whites [relative risk (RR), 1.7; 95% confidence interval (CI), 0.7-3.9]. Registries north of 40 degrees latitude reported higher incidence than registries south of 38 degrees (RR, 3.2; 95% CI, 1.5-7.1). Incidence was higher in women than men (RR, 2.3; 95% CI, 1.2-4.3) and increased steadily with age but did not change over time. Of the 46 (84%) cases with known stage at diagnosis, 37% were confined to the anorectum, 41% had regional spread, and 22% metastasized to distant sites. One and 5-year survivals (+/- SE) were 50% +/- 7% and 15% +/- 6%, respectively. One-year survival for local, regional, and distant disease was 75% +/- 11%, 39% +/- 12%, and 33% +/- 16%, respectively. Prognosis improved over the 15 years of surveillance. Age, sex, race, and registry area were not associated with survival. CONCLUSIONS: Anorectal melanoma differs markedly from cutaneous melanoma in etiology (indeed, sun exposure may be protective) and has a very poor, although improving prognosis. PMID- 8419241 TI - Effect of ethanol on rat gastric surfactant: a fluorescence polarization study. AB - BACKGROUND: Gastric surfactant is believed to protect gastric mucosa by the hydrophobic properties of its phospholipidic component, which are reflected in the fluorescence polarization of a lipophylic fluorescent probe. The present study aimed to observe the consequences of intragastric administration of 40% ethanol on the physical properties of rat gastric surfactant. METHODS: Fluorescence polarization studies and lipid composition of gastric mucosal surface scrapings were performed. RESULTS: Time course experiments indicated that the ulcerogenic action of ethanol occurred along with a fluidization of the surface scrapings followed by secondary rigidification. The fluidizing effect of ethanol was related to modifications of the molecular dynamics of lipid structures. The rigidifying effect of ethanol was a result of an increase in the cholesterol-triglyceride and cholesterol-phospholipid ratios and an increase in the percent composition of phosphatidyl-ethanolamine of surface scrapings. CONCLUSIONS: The results suggest that alcohol could alter the gastric mucosal barrier by its disorganizing effect on the molecular dynamics of the gastric surfactant. The second rigidifying effect of ethanol could be a part of the damage repair phenomenon. PMID- 8419242 TI - Na(+)-dependent and -independent Cl-/HCO3- exchangers in cultured rabbit esophageal epithelial cells. AB - BACKGROUND: The mechanisms by which esophageal epithelial cells regulate intracellular pH (pHi) in a physiological solution are unknown. METHODS: Basal type esophageal cells growing in primary culture were loaded with the fluorescent dye 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) to study pHi by microfluorimetry. RESULTS: The pHi in HEPES buffer was 7.7 +/- 0.03, a value higher than that in CO2/HCO3- buffer, 7.2 +/- 0.1. Cells in HEPES switched to CO2/HCO3- buffer rapidly acidified to pHi of 7, then alkalinized to a new steady state pHi. The mechanisms for alkalinization in CO2/HCO3- were dependent on two exchangers, one amiloride-sensitive and the other 4,4'-diisothiocyanatostilbene 2,2'-disulfonic acid (DIDS)-sensitive, the latter dependent on Nao and Cli, and so indicative of an Na(+)-dependent Cl-/HCO3- exchanger. Cells in a CO2/HCO3- buffer rapidly alkalinized to pH 8.2 when switched to HEPES, then acidified to a new steady-state pHi. Acidification in HEPES was largely caused by a DIDS sensitive, Clo-dependent, non-Nao-requiring mechanism, indicative of a cell acidifying Na-independent Cl-/HCO3- exchanger. CONCLUSIONS: In a physiological buffer, esophageal cells have at least three exchangers for regulation of pHi. PMID- 8419243 TI - Mosaic differentiation of human villus enterocytes: patchy expression of blood group A antigen in A nonsecretors. AB - BACKGROUND: The authors have shown that a mosaicism of brush border antigens may occur spontaneously on enterocytes of small intestine in human adult-type hypolactasia. The present paper gives another example of spontaneously occurring mosaicism as indicated by the patchy expression of blood group antigens on villus enterocytes. METHODS: Thirty-five individuals were examined by immunomorphological techniques with antibodies against blood group antigens. RESULTS: In 4 of 16 A blood group individuals, the blood group antigens were expressed only in some villus enterocytes. The individuals with this mosaic pattern were all shown to be nonsecretors. The A antigen in the positive enterocytes of these individuals was only present as the ALe(b) structure, whereas ALe(y) and ALe(d) were also present in the secretors. The patches of positive enterocytes were randomly distributed along the villus wall. CONCLUSIONS: A nonsecretor individuals express the blood group antigens only in some villus enterocytes; this mosaicism does not arise from a heterogeneous population of stem cells within the crypts but rather reflects subtle differences in the pattern of differentiation between monoclonally derived epithelial cells on the villus. PMID- 8419244 TI - The effect of gamma-aminobutyric acid on hepatic regenerative activity following partial hepatectomy in rats. AB - BACKGROUND: gamma-Aminobutyric acid (GABA) is a potent inhibitory neurotransmitter with growth-regulatory properties. In fulminant hepatic failure, a condition in which hepatic regeneration may be impaired, systemic serum GABA concentrations are markedly elevated. The present study was designed to determine whether increased amounts of circulating GABA interfere with hepatic regenerative activity. METHODS: Exogenous GABA or isotonic saline was administered to adult male rats (n = 6-12/group) 16 hours before partial hepatectomy and twice daily for 1-3 days thereafter. The hypertrophic and hyperplastic components of hepatic regeneration were determined by calculation of the restitution of liver mass, [14C]leucine incorporation into protein (protein synthesis), and [3H]thymidine incorporation into hepatic DNA (DNA synthesis). RESULTS: Exogenous GABA impaired restitution of liver mass (GABA vs. controls, day 3: 76% +/- 7% vs. 90% +/- 9%, mean +/- SD) (P < 0.005) and the rate of protein synthesis (GABA vs. controls, day 1: 379 +/- 39 dpm/mg protein vs. 564 +/- 67 dpm/mg protein) (P < 0.01) without interfering with DNA synthesis. Supplemental administration of corticosterone and putrescine restored protein synthesis rates to normal in GABA treated rats. CONCLUSIONS: These results indicate that elevated serum GABA concentrations interfere with the hypertrophic component of hepatic regeneration following partial hepatectomy in rats. PMID- 8419245 TI - Hemodynamic effects of acute changes in intra-abdominal pressure in patients with cirrhosis. AB - BACKGROUND: Changes in intra-abdominal pressure (IAP) have significant circulatory effects. However, whether this may influence the gastroesophageal collateral blood flow in patients with cirrhosis has not been studied. METHODS: In 14 portal hypertensive cirrhotics, serial hemodynamic measurements were obtained in baseline conditions 30 minutes after the mechanical increase of IAP by 10 mm Hg and 30 minutes after returning IAP to baseline levels. RESULTS: Increasing IAP caused similar increases in free and wedged hepatic venous pressures (+10.3 mm Hg and +11.0 mm Hg, respectively; P < 0.005), without changing the hepatic venous pressure gradient (HVPG). However, there were significant decreases in cardiac output (-18%; P < 0.005) and hepatic blood flow (-20%; P < 0.05), whereas azygos blood flow, an index of gastroesophageal collateral blood flow, increased markedly (+23%; P < 0.005). The opposite occurred after releasing the high IAP. CONCLUSION: In portal hypertensive cirrhotics, acute changes in IAP did not change HVPG but markedly modified splanchnic and systemic hemodynamics. Brief elevations of IAP may have deletereous effects, as shown by the increase in azygos blood flow and the decrease in cardiac output and hepatic blood flow, whereas reduction of a high IAP causes the opposite changes and may be beneficial. PMID- 8419246 TI - Primary liver cancer in genetic hemochromatosis: a clinical, pathological, and pathogenetic study of 54 cases. AB - BACKGROUND: Although liver cancer arises frequently in the course of genetic hemochromatosis (GH), it has not been previously studied in a large series of patients with well-defined GH. METHODS: The bioclinical and pathological data from 1 cholangiocarcinoma and 53 hepatocellular carcinomas (HCCs) complicating GH in 32 untreated and 22 de-ironed patients are reported. RESULTS: This study (1) adds three new well-documented cases of HCC in noncirrhotic but only fibrotic hemochromatotic liver, (2) shows the high prevalence (83%) of proliferative and often dysplastic (70%) iron-free foci in the nontumorous liver of untreated patients, and (3) emphasizes the significant increase of cirrhosis (81% vs. 28%) and of associated noniron-related risk factors, mainly chronic alcoholism (48% vs. 25%) and tobacco smoking (50% vs. 18%) in patients with HCC compared with matched hemochromatotic patients without HCC. CONCLUSIONS: These data (1) suggest that iron-free foci may be markers of an early stage of HCC in GH and (2) supply the basis for defining a cost-effective policy for the screening of HCC in GH patients. PMID- 8419247 TI - Nutritional, hepatic, and metabolic effects of cachectin/tumor necrosis factor in rats receiving total parenteral nutrition. AB - BACKGROUND: In orally fed animals, infusion of cachectin/tumor necrosis factor (TNF) caused weight loss and muscular wasting, accompanied by anorexia. Despite muscle wasting, there were gains in weight and protein and DNA contents of the viscera, but no significant metabolic abnormalities. METHODS: To observe the effect of cachectin/TNF on the nutritional-metabolic status, and without the confounding effect of anorexia, cachectin/TNF was infused into rats receiving total parenteral nutrition in sufficient amounts to induce weight gain in controls at the same rate as in orally fed rats. RESULTS: TPN prevented loss of body weight, but cachectin-treated animals had reduced nitrogen retention and carcass weight. By contrast, there were gains in visceral protein levels, which in the liver was due to a marked proliferation of biliary epithelium. In addition, cachectin-treated animals receiving TPN developed hyperglycemia, hyperosmolality, diuresis, and dehydration. They also had azotemia and cholestasis. CONCLUSIONS: In the absence of the effects of anorexia, cachectin reduced nitrogen retention and caused metabolic and multisystem dysfunction, comparable with the effects of clinical sepsis. PMID- 8419248 TI - Subhypnotic doses of propofol relieve pruritus associated with liver disease. AB - BACKGROUND: Pruritus is a severe and troublesome symptom in patients with cholestasis and is often difficult to treat. Propofol was recently shown to be efficient in relieving pruritus secondary to spinal morphine administration. The efficacy of propofol was therefore investigated in patients with pruritus associated with liver disease. METHODS: In a prospective, randomized, double blind, crossover, placebo-controlled study, 10 patients received 2 doses of propofol (1.5 mL = 15 mg) and 2 doses of placebo (1.5 mL of Intralipid, Kabi Pharm., Helsinki, Finland) during a 4-day study period. Pruritus was assessed by a verbal rating score from 0 (no pruritus) to 10 (most severe pruritus imaginable). Treatment success was defined as a decrease of pruritus of at least 4 points in the verbal rating score. RESULTS: Treatment success was achieved in 85% of the patients receiving propofol and in 10% of those receiving Intralipid (P < 0.01). Discomfort on injection (15%) and slight dizziness (10%) were observed with propofol treatment. CONCLUSIONS: This study shows that subhypnotic doses of propofol are effective for the short-term symptomatic relief of pruritus associated with liver disease. At the dose used, side effects were rare and minor. PMID- 8419249 TI - The role of prostaglandin I2 and biliary lipids during evolving cholecystitis in the rabbit. AB - BACKGROUND: Acute cholecystitis increases gallbladder prostanoid synthesis. The percent study examined the hypothesis that increased endogenous gallbladder release of prostaglandin I2 (PGI2) after bile duct ligation is caused by both increased ductal pressure and altered biliary lipids. METHODS: Prostanoid release, biliary lipids, and in vitro fluid absorption of sham gallbladders were compared with those of gallbladders in which acute cholecystitis was induced after common bile duct ligation for 6, 24, and 72 hours. RESULTS: Bile duct ligation for 6, 24, and 72 hours increased gallbladder PGI2 release twofold and increased gallbladder bile levels of lysolecithin and taurine-conjugated bile acids fivefold compared with sham groups (P < 0.05). In vitro gallbladder fluid absorption was decreased by 50% or more in the 6-, 24-, and 72-hour bile duct ligated groups (P < 0.05) but was reversed by indomethacin only in the 6-hour ligated group. CONCLUSIONS: Decreased gallbladder fluid absorption following bile duct ligation for 6 hours was caused by increased gallbladder release of PGI2. Decreased gallbladder fluid absorption following bile duct ligation for 24 and 72 hours was not a prostanoid-mediated process (not reversed by indomethacin) but was associated with increased bile levels of proinflammatory biliary lipids. PMID- 8419250 TI - Precore mutations and core clustering mutations in chronic hepatitis B virus infection. AB - BACKGROUND: Mutant hepatitis B virus is often associated with severe liver damage. The purpose of this study is to elucidate the relationship between mutations in hepatitis B precore/core gene and the severity of liver damage. METHODS: The hepatitis B precore/core gene from 20 patients with chronic hepatitis B virus infection was studied by polymerase chain reaction and direct sequencing. RESULTS: Missense mutations in the core gene were only found in patients with chronic active hepatitis. Three mutation clustering regions of core gene, codons 48-60, 84-101, and 147-155, had higher substitution rates than other regions. All patients with chronic active hepatitis had missense mutation(s) either in codons 84-101 or in codons 48-60. There was a trend of increasing substitutions in the precore/core gene from e antigen-positive asymptomatic carriers to e antibody-positive patients with chronic active hepatitis. CONCLUSIONS: These data suggest that (1) severe liver damage in chronic hepatitis B virus infection is related to the clustering missense mutations in codons 48-60 and 84-101 of core gene and that (2) the emergence of precore stop codon mutation and missense mutations around the carboxy-terminal processing site of precore/core protein (codons 147-155) may be the adaptive mechanisms of hepatitis B virus to decrease production and secretion of viral protein and retain the viral persistence. PMID- 8419251 TI - Alpha-chain disease: analysis of alpha-chain protein and secretory component in jejunal fluid. AB - BACKGROUND: It is unclear why different forms of alpha-chain disease protein appear in intestinal fluid. This was studied in a 23-year-old Mauritanian man in whom alpha-chain disease was diagnosed localized to the duodenum and jejunum, nasopharynx, and bone marrow. METHODS: The duodenal infiltrate was studied by immunohistochemistry. Forms of alpha chain-containing proteins in serum and jejunal fluid were analyzed by ultracentrifugation and radioimmunoassays. RESULTS: The infiltrating cells contained alpha-1 chain but no light chains, and approximately 66% showed variable expression of J chain. Serum contained a large fraction of monomeric alpha-chain disease protein, whereas both monomeric and heavier forms appeared in jejunal fluid. Some of the latter were bound to secretory component, and the fluid contained virtually no free component. CONCLUSIONS: Linkage of polymeric alpha-chain disease protein to secretory component depends on balanced synthesis of alpha chains and J chain in the proliferating B cells, giving rise to polymers with binding site for secretory component expressed as an epithelial receptor. Insufficient receptor-mediated transport capacity (either relative and/or because of intestinal crypt reduction) results in passive external transfer of polymers without bound secretory component along with leakage of serum-derived or locally produced monomeric alpha chain disease protein, the latter presumably originating from immunocytes with little or no J-chain synthesis. PMID- 8419253 TI - Tempora mutantur, nos et mutamur in illis! PMID- 8419252 TI - Hepatobiliary complications of total parenteral nutrition. AB - The relationships between various hepatobiliary disorders and the administration of total parenteral nutrition (TPN) were reviewed and, in particular, the role of TPN in their pathogenesis was critically evaluated. Several clinical and pathological entities including steatosis, steatohepatitis, cholestasis, and cholelithiasis have been commonly linked to TPN, and instances of chronic decompensated liver disease have been reported. However, it is concluded that it is often difficult to extricate the effects of TPN on hepatobiliary function from many other hepatotoxic factors that may be operative in these patients. Thus, whereas considerable evidence exists to support a role fro carbohydrate or calorie excess in TPN solutions in the pathogenesis of steatosis, a loss of enteric stimulation and not TPN per se may be the primary factor in the development of cholestasis, biliary sludge, and gallstones. The apparent predilection of infants to TPN-related cholestasis may be based on the relative immaturity of the neonatal biliary excretory system. PMID- 8419254 TI - A 19-year-old woman with unexplained vomiting. PMID- 8419255 TI - Modulation by prostanoids of the release of inflammatory mediators from mast cells: involvement in mucosal protection? PMID- 8419256 TI - Postoperative feeding: liquid versus solid fuel. PMID- 8419257 TI - Purine analogues and ulcerative colitis. PMID- 8419258 TI - An enteric pathogen subverts epithelial signal transduction mechanisms for invasion. PMID- 8419259 TI - Does the gallbladder secrete? PMID- 8419260 TI - Negative feedback regulation of the ileal bile acid transport system in rodents. AB - BACKGROUND: Active transport of conjugated bile acids by ileal enterocytes is a key mechanism for conservation of the bile acid pool. Experiments were performed to determine whether such transport is regulated by substrate load. METHODS: Using anesthetized biliary fistula guinea pigs or rats, the ileum was perfused with ursodeoxycholyltaurine at a concentration causing maximal ileal transport of this bile acid; absorption was assessed by biliary recovery. Before ileal perfusion, animals ingested one of three diets: chow, chow with added conjugated bile acid, or chow with added cholestyramine. RESULTS: In the guinea pig, ingestion of a taurocholate-enriched diet resulted in a 75% decrease in the absorption rate of ursodeoxycholyltaurine. Similar results were obtained with cholylsarcosine (a deconjugation-dehydroxylation resistant analogue) or with chenodeoxycholylglycine, the endogenous bile acid of the guinea pig. In contrast, cholestyramine ingestion caused an increase in ursodeoxycholyltaurine absorption. In the rat, cholyltaurine or cholylsarcosine ingestion also caused decreased ileal transport. In the guinea pig, maximal down-regulation of active ileal bile acid transport occurred after 2-3 days of bile acid feeding; up-regulation required 3-4 days. CONCLUSIONS: Bile acid metabolism is regulated by feedback inhibition of active ileal transport in addition to the well-established feedback inhibition of bile acid biosynthesis in the liver. Together, these two regulatory mechanisms ensure constancy of bile acid secretion. PMID- 8419261 TI - Platelet-activating factor and interleukin 1 are involved in colonic dysmotility in experimental colitis in rats. AB - BACKGROUND: Intracolonic administration of trinitrobenzene sulfonic acid (TNBS) to rats produces chronic colitis associated with an increased release of eicosanoids, platelet-activating factor (PAF), and interleukins. METHODS: Motor effects of TNBS on proximal colon were evaluated electromyographically in rats. Mediator involvement was investigated using eicosanoids and PAF antagonists. RESULTS: The colonic myoelectrical activity was 59 +/- 17 spike bursts per hour lasting 6.9 +/- 1.3 seconds. Two to eight days after TNBS treatment, spike-burst duration was significantly (P < 0.05) higher, with a maximal 1.5-4-fold enhancement at day 3. These alterations were significantly (P < 0.05) reduced by daily treatment with MK-886, a 5-lipoxygenase inhibitor (10 mg/kg, orally), whereas indomethacin (1 mg/kg per day, intramuscularly) was ineffective. At day 3, RP55778, a PAF antagonist (45, 60 mg/kg, intraperitoneally), and rIRAP, an interleukin 1 antagonist (0.3 mg/kg, intraperitoneally) but not KT1-32, a thromboxane A2 antagonist (30, 60 mg/kg orally), nor SKF104,353, a leukotriene D4 antagonist (2, 4 mg/kg, orally), significantly (P < 0.05) reduced the TNB-induced motor effects. CONCLUSION: TNBS-induced colitis in rats involves a delayed long lasting dysmotility involving PAF, interleukin 1, and some leukotrienes but not leukotriene D4, thromboxane A2, or other cyclo-oxygenase products. PMID- 8419262 TI - Prognostic value of p53 overexpression and c-Ki-ras gene mutations in colorectal cancer. AB - BACKGROUND: Mutations in Ki-ras codon 12 and the p53 gene are common abnormalities in colorectal cancer. The occurrence of p53 overexpression and/or Ki-ras codon 12 mutations were analyzed in 100 colorectal adenomas to determine if they were related to patient survival. METHODS: p53 overexpression was identified by immunohistochemistry, and Ki-ras codon 12 mutations were detected using the polymerase chain reaction and a restriction enzyme digestion method. RESULTS: p53 overexpression was identified in 45% of tumors, with a higher frequency identified in DNA aneuploid and left-sided tumors than in DNA diploid and right-sided tumors. Mutations in Ki-ras codon 12 were identified in 24% of carcinomas. Individually, mutations in Ki-ras codon 12 or p53 overexpression were not prognostic indicators of survival. However, a statistically significant difference in survival was identified when these two oncogenic abnormalities were analyzed together. The median survival of patients whose tumors contained both oncogenic abnormalities was less than half of that of patients with either alteration alone or without either abnormality. CONCLUSIONS: Screening for multiple genetic abnormalities in colorectal cancers excised at surgery may prove to be a useful tool in determining prognosis. PMID- 8419263 TI - The specialization of gastroenterology in America. PMID- 8419264 TI - Transport of the biotin dietary derivative biocytin (N-biotinyl-L-lysine) in rat small intestine. AB - BACKGROUND: Biocytin is an important end product of intraluminal digestion of dietary protein-bound biotin. Limited studies are available regarding the ability of the small intestine to transport biocytin and about the mechanism involved. The aim of the present study was to delineate these issues. METHODS: Transport of [3H]-biocytin was examined using everted sacs from rat intestine. RESULTS: Mucosal-to-serosal transport of low (0.022 mumol/L) and high (5 mumol/L) concentrations of biocytin were linear for up to 20 minutes of incubation. Transport of biocytin as a function of concentration (0.022-5 mumol/L) was linear (r = 0.99) and occurred at a rate of 22,062 fmol.g tissue (wet wt)-1.15 min-1. Addition of high concentrations of unlabeled biocytin, biotin, biotin methyl ester, and lysine did not cause a significant inhibition of the transport of [3H] biocytin. Furthermore, transport of biocytin was independent of Na+ concentration, pH, energy, and temperature. Compared with transport of equimolar concentrations of free biotin, transport of biocytin (0.022 mumol/L) was significantly lower in both the jejunum and the ileum. CONCLUSIONS: (1) Biocytin transport in rat intestine is lower than that of free biotin and occurs via simple physical diffusion. (2) In the rat, efficient absorption and optimal bioavailability of dietary protein-bound biotin necessitates its conversion to free biotin. PMID- 8419265 TI - Absorption of glucose polymers from rice in oral rehydration solutions by rat small intestine. AB - BACKGROUND: This study aims to determine the effect of replacing glucose in oral rehydration solution (ORS) with starch hydrolysates from rice on absorption in the small intestine and levels of glucose in portal venous blood and on disaccharidase levels and morphometric measurements in intestines of rats. METHODS: ORS containing standard composition of salts and 2% glucose (WHO ORS) or 2%, 5%, or 10% starch hydrolysates were infused into duodena of 60 Sprague-Dawley rats (250-350 g). Portal venous blood glucose levels were determined at 0, 30, 60, 90, and 120 minutes. RESULTS: Significantly larger areas under the curve of glucose absorption (AUCs) were produced by ORS containing 10% unfractionated starch hydrolysates (123.2 +/- 3.8), 2%, 5%, and 10% starch hydrolysates with long-chain ( > 9 molecules) glucose polymers (109.5 +/- 10.6, 109.3 +/- 7.4, and 115.3 +/- 7.1, respectively), and 5% and 10% starch hydrolysates with short-chain (2-9 molecules) glucose polymers (124.4 +/- 6.1 and 128.1 +/- 6.8). ORS with 2% and 5% unfractionated starch hydrolysates and 2% short-chain glucose polymers produced AUCs comparable with those of WHO ORS (96.48 +/- 5.7). Disaccharidase levels and morphometric measurements were not significantly different. CONCLUSIONS: Starch hydrolysates from rice containing glucose polymers can be used in ORS in higher concentrations than glucose to provide higher caloric density without increased osmolality. PMID- 8419266 TI - TB returns: an opportunity lost. PMID- 8419267 TI - Infections in older patients: a systematic clinical approach. AB - The increased risk of infection among older patients can be divided into three clinical categories: infections increased in incidence, infections showing higher case fatality rates, and infections that are clinically worse, primarily because of late recognition. Among infections that are increased in incidence, the most important by far are tuberculosis and pneumococcal pneumonia. Infections that show higher fatality rates include influenza and--again--pneumococcal pneumonia. Intra-abdominal infections (eg, cholecystitis and appendicitis) are often clinically worse in older patients due to late recognition and delay in surgical intervention. PMID- 8419268 TI - Pressure ulcers: updated guidelines for treatment and prevention. AB - Pressure ulcers are prevalent and associated with high rates of morbidity and mortality in older patients. Four primary forces--pressure, shearing, friction, and moisture--contribute to their formation. A number of factors place patients at risk, including immobility, age-related skin changes, malnutrition, and cognitive impairment. Turning schedules, special bed surfaces, and other preventive measures have been shown to significantly reduce the effects of these factors. When an ulcer does form, treatment is based on the degree of tissue damage and includes pressure-reducing devices, systemic measures, and local care with debridement, occlusive dressings, and sometimes surgery. PMID- 8419269 TI - The role of dipyridamole in the therapy of vascular disease. AB - The antiplatelet agent dipyridamole is FDA-approved as an adjunct to warfarin for the prevention of thromboembolism in patients receiving prosthetic heart values. It is also prescribed in several other situations, although data supporting these uses remain equivocal. Dipyridamole does not appear to be superior to aspirin alone in the management of patients with cerebral or coronary artery disease nor in maintaining the patency of autologous grafts. Its role in the management of patients who fail aspirin or other antithrombotic therapy remains to be examined. The drug may offer long-term advantages in the prevention of atherogenesis, but this use also warrants future investigation. PMID- 8419270 TI - Older patients with COPD: benefits of exercise training. AB - Chronic obstructive pulmonary disease (COPD) is a leading cause of mortality and functional disability in older adults. In its advanced stages, it is characterized by progressive breathlessness and serious exercise curtailment. Aerobic exercise training (EXT) is recommended for patients who remain breathless despite optimal pharmacologic treatment. Although the physiologic rationale for EXT in COPD patients remains controversial, it is generally accepted that even older incapacitated COPD sufferers who participate in an individualized training program show significant reductions in breathlessness, increased exercise capacity, and enhanced psychosocial function. An 8-week outpatient program that is carefully regulated and includes an educational component is recommended. PMID- 8419271 TI - Thirteen views from the Hill. Congressional leaders paint varying reform scenarios. Interview by Marybeth Burke. PMID- 8419272 TI - Networks and our next step. Communicating the hospital community's vision on reform. PMID- 8419273 TI - Clinton's next moves. Scoping out the White House on health care reform. PMID- 8419274 TI - The new Congress. What happens to health care reform? PMID- 8419275 TI - Reach out and heal someone. Health care workers aid beleaguered hospitals in former Soviet republics. PMID- 8419276 TI - CEOs in the 1990s. Executive careers will focus on interpersonal skills. Interview by Mary Grayson. PMID- 8419277 TI - Change is not an option. Spurred by health care reform, boards revamp their roles. PMID- 8419278 TI - Hospitals, vendors explore equipment financing. AB - An uncertain reimbursement environment--clouded by such immediate concerns as Medicare capital payments and longer-term prospects for comprehensive health care reform--is leading top imaging equipment manufacturers to offer hospitals creative ways to finance equipment. PMID- 8419280 TI - Hospital customizes software to achieve internal needs. PMID- 8419279 TI - Third-quarter data from AHA Monitrend II. PMID- 8419281 TI - Austin decision good news for peer reviewers. PMID- 8419282 TI - Slowing rate of cost increases in the pharmacy. AB - As the emergence of new drugs and technology and the growing acuity of patients drive up the cost of hospital pharmaceuticals, hospital administrators can act to slow the rate of increase. The key, writes the author, lies in recognizing that newness in pharmaceuticals does not equal clinical superiority or cost effectiveness. PMID- 8419283 TI - Autogenous regulation of gene expression. PMID- 8419284 TI - Cloning and characterization of the ferric enterobactin receptor gene (pfeA) of Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa K407, a mutant lacking a high-affinity 80,000-molecular weight ferric enterobactin receptor protein (80K protein), exhibited poor growth (small colonies) on iron-deficient succinate minimal medium containing ethylenediamine-di(o-hydroxyphenylacetic acid) (EDDHA) and enterobactin. The gene encoding the ferric enterobactin receptor was cloned by complementation of this growth defect. The complementing DNA was subsequently localized to a 7.1-kilobase pair (kb) SstI-HindIII fragment which was able to restore synthesis of the 80K protein in strain K407 and also to direct the synthesis of high levels of a protein of the same molecular weight in the outer membranes of Escherichia coli fepA strains MT912 and IR20. Moreover, the fragment complemented the fepA mutation in MT912, restoring both growth in EDDHA-containing medium and enterobactin-dependent uptake of 55Fe3+. Expression of the P. aeruginosa receptor in E. coli IR20 was shown to be regulated by both iron and enterobactin. The complementing DNA was further localized to a 5.3-kb SphI-SstI fragment which was then subjected to deletion analysis to obtain the smallest fragment capable of directing the synthesis of the 80K protein in the outer membrane of strain K407. A 3.2-kb DNA fragment that restored production of the receptor in strain K407 was subsequently isolated. The fragment also directed synthesis of the protein in E. coli MT912 but at levels much lower than those previously observed. Nucleotide sequencing of the fragment revealed an open reading frame (designated pfeA for Pseudomonas ferric enterobactin) of 2,241 bp capable of encoding a 746-amino-acid protein with a molecular weight of 80,967. The PfeA protein showed more than 60% homology to the E. coli FepA protein. Consistent with this, the two proteins showed significant immunological cross-reactivity. PMID- 8419285 TI - Characterization of DNA helicase II from a uvrD252 mutant of Escherichia coli. AB - The loss of DNA helicase II (UvrD) in Escherichia coli results in sensitivity to UV light and increased levels of spontaneous mutagenesis. While the effects of various uvrD alleles have been analyzed in vivo, the proteins produced by these alleles have not been examined in any detail. We have cloned one of these alleles, uvrD252, and determined the site of the mutation conferring the phenotype. In addition, the protein it encodes has been purified to homogeneity and characterized in vitro. The mutation responsible for the phenotype was identified as a glycine-to-aspartic-acid change in the putative ATP-binding domain. In comparison to wild-type DNA helicase II, the UvrD252 enzyme exhibited reduced levels of ATPase activity and a large increase in the Km for ATP. The ability of UvrD252 to unwind DNA containing single-stranded regions, as well as DNA containing only nicks, was reduced in comparison to that of the wild-type enzyme. Possible interpretations of these results in relation to the phenotypes of the uvrD252 mutant are discussed. This represents the first detailed analysis of the biochemical properties of a mutant DNA helicase II protein. PMID- 8419286 TI - Complementation between nucleotide binding domains in an anion-translocating ATPase. AB - The catalytic component of the oxyanion-translocating ATPase of the plasmid encoded ars operon of Escherichia coli is a homodimer of the ArsA protein. This enzyme is an oxyanion-stimulated ATPase with two consensus nucleotide binding sequences in each subunit, one in the N-terminal (A1) half and one in the C terminal (A2) half of the ArsA protein. The two halves of both the arsA gene and the ArsA protein exhibit similar nucleotide and amino acid sequences, respectively. The two halves of the arsA gene were subcloned into compatible plasmids. Neither alone was sufficient to confer resistance, but cells in which the arsA1 and arsA2 half genes were coexpressed were resistant to arsenicals. Genetic complementation was also observed in cells bearing plasmids with point mutations in the two halves of the arsA gene and between cells with plasmids carrying combinations of the arsA1 or arsA2 subclones and point mutations. In every case, complementation was observed only when one plasmid contained a wild type arsA1 sequence and the other contained a wild-type arsA2 sequence. These results demonstrate that both sites are required for resistance but that the two nucleotide binding domains need not reside in a single polypeptide. We propose a model in which the ArsA dimer has two catalytic units, each composed of an A1 domain from one monomer and an A2 domain from the other monomer. PMID- 8419287 TI - Identification of the promoter and a negative regulatory element, ftr4, that is needed for cell cycle timing of fliF operon expression in Caulobacter crescentus. AB - The fliF operon of Caulobacter crescentus, which was previously designated the flaO locus, is near the top of the flagellar-gene regulatory hierarchy, and it is one of the earliest transcription units to be expressed in the cell cycle. In this report, we have identified two cis-acting sequences that are required for cell cycle regulation of fliF transcription. The first sequence was defined by the effects of three 2-bp deletions and five point mutations, each of which greatly reduced the level of fliF operon transcript in vivo. These eight mutations lie between -37 and -22 within an 18-bp sequence that matches, at 11 nucleotides, sequences in the 5' regions of the flaQR (flaS locus) and fliLM operons, which are also expressed early and occupy a high level in the regulatory hierarchy (A. Dingwall, A. Zhuang, K. Quon, and L. Shapiro, J. Bacteriol. 174:1760-1768, 1992). We propose that this 18-bp sequence contains all or part of the fliF promoter. We have also identified a second sequence, 17 bp long and centered at -8, which we have provisionally designated ftr4 because of its similarity to the enhancer-like ftr sequences required for regulation of sigma 54 promoters flaN and flbG (D. A. Mullin and A. Newton, J. Bacteriol. 171:3218-3227, 1989). Six of the seven mutations in ftr4 examined resulted in a large increase in fliF operon transcript levels, suggesting a role for ftr4 in negative regulation. A 2-bp deletion at -12 and -13 in ftr4 altered the cell cycle pattern of fliF operon transcription; the transcript was still expressed periodically, but the period of its synthesis was extended significantly. We suggest that the ftr4 sequence may form part of a developmental switch which is required to turn off fliF operon transcription at the correct time in the cell cycle. PMID- 8419288 TI - Purification and properties of the physically associated meta-cleavage pathway enzymes 4-hydroxy-2-ketovalerate aldolase and aldehyde dehydrogenase (acylating) from Pseudomonas sp. strain CF600. AB - The final two steps in the dmp operon-encoded meta-cleavage pathway for phenol degradation in Pseudomonas sp. strain CF600 involve conversion of 4-hydroxy-2 ketovalerate to pyruvate and acetyl coenzyme A (acetyl-CoA) by the enzymes 4 hydroxy-2-ketovalerate aldolase and aldehyde dehydrogenase (acylating) [acetaldehyde:NAD+ oxidoreductase (CoA acetylating), EC 1.2.1.10]. A procedure for purifying these two enzyme activities to homogeneity is reported here. The two activities were found to copurify through five different chromatography steps and ammonium sulfate fractionation, resulting in a preparation that contained approximately equal proportions of two polypeptides with molecular masses of 35 and 40 kDa. Amino-terminal sequencing revealed that the first six amino acids of each polypeptide were those deduced from the previously determined nucleotide sequences of the corresponding dmp operon-encoded genes. The isolated complex had a native molecular mass of 148 kDa, which is consistent with the presence of two of each polypeptide per complex. In addition to generating acetyl-CoA from acetaldehyde, CoA, and NAD+, the dehydrogenase was shown to acylate propionaldehyde, which would be generated by action of the meta-cleavage pathway enzymes on the substrates 3,4-dimethylcatechol and 4-methylcatechol. 4-Hydroxy-2 ketovalerate aldolase activity was stimulated by the addition of Mn2+ and, surprisingly, NADH to assay mixtures. The possible significance of the close physical association between these two polypeptides in ensuring efficient metabolism of the short-chain aldehyde generated by this pathway is discussed. PMID- 8419289 TI - The Saccharomyces cerevisiae SPR1 gene encodes a sporulation-specific exo-1,3 beta-glucanase which contributes to ascospore thermoresistance. AB - A number of genes have been shown to be transcribed specifically during sporulation in Saccharomyces cerevisiae, yet their developmental function is unknown. The SPR1 gene is transcribed during only the late stages of sporulation. We have sequenced the SPR1 gene and found that it has extensive DNA and protein sequence homology to the S. cerevisiae EXG1 gene which encodes an exo-1,3-beta glucanase expressed during vegetative growth (C. R. Vasquez de Aldana, J. Correa, P. San Segundo, A. Bueno, A. R. Nebrada, E. Mendez, and F. del Ray, Gene 97:173 182, 1991). We show that spr1 mutant cells do not hydrolyze p-nitrophenyl-beta-D glucoside or laminarin in a whole-cell assay for exo-1,3-beta-glucanases. In addition to the absence of this enzymatic activity, spr1 mutant spores exhibit reduced thermoresistance relative to isogenic wild-type spores. These observations are consistent with the notion that SPR1 encodes a sporulation specific exo-1,3-beta-glucanase. PMID- 8419290 TI - Oxidation of biphenyl by a multicomponent enzyme system from Pseudomonas sp. strain LB400. AB - Pseudomonas sp. strain LB400 grows on biphenyl as the sole carbon and energy source. This organism also cooxidizes several chlorinated biphenyl congeners. Biphenyl dioxygenase activity in cell extract required addition of NAD(P)H as an electron donor for the conversion of biphenyl to cis-2,3-dihydroxy-2,3 dihydrobiphenyl. Incorporation of both atoms of molecular oxygen into the substrate was shown with 18O2. The nonlinear relationship between enzyme activity and protein concentration suggested that the enzyme is composed of multiple protein components. Ion-exchange chromatography of the cell extract gave three protein fractions that were required together to restore enzymatic activity. Similarities with other multicomponent aromatic hydrocarbon dioxygenases indicated that biphenyl dioxygenase may consist of a flavoprotein and iron-sulfur proteins that constitute a short electron transport chain involved in catalyzing the incorporation of both atoms of molecular oxygen into the aromatic ring. PMID- 8419291 TI - Nucleotide sequence and initial functional characterization of the clcR gene encoding a LysR family activator of the clcABD chlorocatechol operon in Pseudomonas putida. AB - The 3-chlorocatechol operon clcABD is central to the biodegradative pathway of 3 chlorobenzoate. The clcR regulatory gene, which activates the clcABD operon, was cloned from the region immediately upstream of the operon and was shown to complement an insertion mutation for growth on 3-chlorobenzoate. ClcR activated the clcA promoter, which controls expression of the clcABD operon, in trans by 14 fold in an in vivo promoter probe assay in Pseudomonas putida when cells were incubated with 15 mM 3-chlorobenzoic acid. Specific binding of ClcR to the clcR clcA intergenic promoter region was observed in a gel shift assay. Nucleotide sequence analysis of the clcR gene predicts a polypeptide of 32.5 kDa, which was confirmed by using specific in vivo 35S labeling of the protein from a T7 promoter-controlled ATG fusion construct. ClcR shares high sequence identity with the LysR family of bacterial regulator proteins and has especially high homology to a subgroup of the family consisting of TcbR (57% amino acid sequence identity), TfdS, CatR, and CatM. ClcR was shown to autoregulate its own production in trans to 35% of unrepressed levels but partially relieved this autorepression under conditions that induced transcription at the clcA promoter. Several considerations indicate that the clcR-clcABD locus is most similar to the tcbR-tcbCDEF regulon. PMID- 8419292 TI - Suppression of ColE1 high-copy-number mutants by mutations in the polA gene of Escherichia coli. AB - We isolated three Escherichia coli suppressor strains that reduce the copy number of a mutant ColE1 high-copy-number plasmid. These mutations lower the copy number of the mutant plasmid in vivo up to 15-fold; the wild-type plasmid copy number is reduced by two- to threefold. The suppressor strains do not affect the copy numbers of non-ColE1-type plasmids tested, suggesting that their effects are specific for ColE1-type plasmids. Two of the suppressor strains show ColE1 allele specific suppression; i.e., certain plasmid copy number mutations are suppressed more efficiently than others, suggesting specificity in the interaction between the suppressor gene product and plasmid replication component(s). All of the mutations were genetically mapped to the chromosomal polA gene, which encodes DNA polymerase I. The suppressor mutational changes were identified by DNA sequencing and found to alter single nucleotides in the region encoding the Klenow fragment of DNA polymerase I. Two mutations map in the DNA-binding cleft of the polymerase region and are suggested to affect specific interactions of the enzyme with the replication primer RNA encoded by the plasmid. The third suppressor alters a residue in the 3'-5' exonuclease domain of the enzyme. Implications for the interaction of DNA polymerase I with the ColE1 primer RNA are discussed. PMID- 8419293 TI - Multiple copies of nodD in Rhizobium tropici CIAT899 and BR816. AB - Rhizobium tropici strains are able to nodulate a wide range of host plants: Phaseolus vulgaris, Leucaena spp., and Macroptilium atropurpureum. We studied the nodD regulatory gene for nodulation of two R. tropici strains: CIAT899, the reference R. tropici type IIb strain, and BR816, a heat-tolerant strain isolated from Leucaena leucocephala. A survey revealed several nodD-hybridizing DNA regions in both strains: five distinct regions in CIAT899 and four distinct regions in BR816. Induction experiments of a nodABC-uidA fusion in combination with different nodD-hybridizing fragments in the presence of root exudates of the different hosts indicate that one particular nodD copy contributes to nodulation gene induction far more than any other nodD copy present. The nucleotide sequences of both nodD genes are reported here and show significant homology to those of the nodD genes of other rhizobia and a Bradyrhizobium strain. A dendrogram based on the protein sequences of 15 different NodD proteins shows that the R. tropici NodD proteins are linked most closely to each other and then to the NodD of Rhizobium phaseoli 8002. PMID- 8419294 TI - The Salmonella typhimurium nadC gene: sequence determination by use of Mud-P22 and purification of quinolinate phosphoribosyltransferase. AB - The Salmonella typhimurium nadC gene and its product, quinolinic acid phosphoribosyltransferase (QAPRTase), were characterized at the molecular and biochemical levels. Fusions of Mud-lac elements isolated in the nadC gene were converted to Mud-P22 insertions. Starting with six original Mud-lac fusions, the entire sequence of the nadC gene was readily obtained. The sequence shows a long open reading frame with two potential initiator methionines, one of which is preceded by the Shine-Dalgarno sequence GGAG-7-nucleotide-ATG. The protein predicted from this second open reading frame is 297 residues in length. The nadC gene was subcloned into a T7-based expression system, allowing for facile purification of the QAPRTase (EC 2.4.2.19) protein to homogeneity. Upon gel filtration, the protein gave an M(r) of 72,000, and sodium dodecyl sulfate polyacrylamide gel electrophoresis gave a subunit M(r) of 35,000. Automated Edman degradation of several tryptic peptides confirmed the amino acid sequence predicted from the DNA sequence. Chromatography of the apparently homogeneous enzyme on reverse-phase high-performance liquid chromatography resolved two protein species. One of these species failed to give an amino-terminal sequence, while the other yielded the amino-terminal sequence predicted by the second open reading frame and lacked the initiator methionine. The mass of the mature protein, predicted from its DNA sequence, was 32,428 Da. Electrospray mass spectrometry gave masses of 32,501 and 32,581 Da for the two peptides. Steady state kinetics on the purified QAPRTase indicated Km values of 32 microM for 5 phosphoribosyl-1-pyrophosphate and 20 microM for quinolinate. Vmax was 0.9 U/mg, similar to values reported for this enzyme by other sources. PMID- 8419295 TI - Structures of and allelic diversity and relationships among the major outer membrane protein (ompA) genes of the four chlamydial species. AB - DNA sequences coding for 81% of the ompA gene from 24 chlamydial strains, representing all chlamydial species, were determined from DNA amplified by polymerase chain reactions. Chlamydial strains of serovars and strains with similar chromosomal restriction fragment length polymorphism had identical ompA DNA sequences. The ompA sequences were segregated into 23 different ompA alleles and aligned with each other, and phylogenetic relationships among them were inferred by neighbor-joining and maximum parsimony analyses. The neighbor-joining method produced a single phylogram which was rooted at the branch between two major clusters. One cluster included all Chlamydia trachomatis ompA alleles (trachoma group). The second cluster was composed of three major groups of ompA alleles: psittacosis group (alleles MN, 6BC, A22/M, B577, LW508, FEPN, and GPIC), pneumonia group (Chlamydia pneumoniae AR388 with the allele KOALA), and polyarthritis group (ruminant and porcine chlamydial alleles LW613, 66P130, L71, and 1710S with propensity for polyarthritis). These groups were distinguished through specific DNA sequence signatures. Maximum parsimony analysis yielded two equally most parsimonious phylograms with topologies similar to the ompA tree of neighbor joining. Two phylograms constructed from chlamydial genomic DNA distances had topologies identical to that of the ompA phylogram with respect to branching of the chlamydial species. Human serovars of C. trachomatis with essentially identical genomes represented a single taxonomic unit, while they were divergent in the ompA tree. Consistent with the ompA phylogeny, the porcine isolate S45, previously considered to be Chlamydia psittaci, was identified as C. trachomatis through biochemical characteristics. These data demonstrate that chlamydial ompA allelic relationships, except for human serovars of C. trachomatis, are cognate with chromosomal phylogenies. PMID- 8419296 TI - Two tRNA gene clusters associated with rRNA operons rrnD and rrnE in Bacillus subtilis. AB - Sequence analysis of cloned rescued DNA fragments from a Bacillus subtilis strain with an inserted recombinant plasmid in ribosomal operon rrnE revealed the presence of two tRNA genes for Met and Asp at the 3' end of the operon. Probing chromosomal DNA from a strain carrying a plasmid inserted in rrnD with a fragment containing the genetically unassigned cluster of 16 tRNA genes revealed that the cluster is located immediately following the rrnD operon. Our findings show that all 10 rrn operons in B. subtilis are associated with tRNA gene clusters. PMID- 8419297 TI - Genetic determinants of host ranges of Bacillus sphaericus mosquito larvicidal toxins. AB - The 51.4-kDa-41.9-kDa binary toxin produced by different strains of Bacillus sphaericus shows differential activity toward Culex quinquefasciatus, Aedes atropalpus, and Aedes aegypti mosquito larvae. The patterns of larvicidal activity toward all three mosquito species and growth retardation in A. aegypti have been shown to be due to the 41.9-kDa protein. By using mutant toxins expressed in Escherichia coli, insecticidal activity and growth retardation correlated with amino acids centered around position 100 of the 41.9-kDa protein. In its response to these toxins, A. atropalpus resembled C. quinquefasciatus rather than its congener, A. aegypti. PMID- 8419298 TI - Repressor binding to a regulatory site in the DNA coding sequence is sufficient to confer transcriptional regulation of the vir-repressed genes (vrg genes) in Bordetella pertussis. AB - Five TnphoA fusions to vir-repressed genes (vrg genes) have been identified in the respiratory pathogen Bordetella pertussis. A comparison of vrg DNA sequences suggests a consensus DNA element within the coding regions of four of five vrg genes. To determine the role of this DNA sequence in vrg regulation, a nucleotide substitution mutation in the conserved region of vrg-6 was isolated. This mutant showed constitutively high levels of expression in the absence of antigenic modulators, MgSO4 and nicotinic acid, suggesting that this DNA element may be a control site for vrg repression. Moreover, Northern (RNA) analysis and transcriptional fusion analysis suggest that vrg genes are regulated at the transcriptional level. To determine whether sequences in the coding region were sufficient to respond to antigenic modulation, a vrg-6::TnphoA promoter deletion plasmid that contained a heterologous promoter driving the expression of vrg-6 coding sequences from the vrg-6 translation start site to the TnphoA fusion junction was constructed. This heterologous construct responded to modulators in a vir-dependent fashion, indicating that sequences upstream of the coding sequence are not required for antigenic modulation. Southwestern (DNA-protein) analysis and mutational studies suggest that the vrg consensus DNA sequence is specifically recognized by a 34-kDa vir-activated gene (vag) product, whose binding results in down-regulation of vrg transcript levels. We conclude, at least for the vrg::TnphoA fusion strains, that a site on the DNA that corresponds to a consensus sequence located in the vrg coding region is sufficient to confer the transcriptional regulation (repression) of vrg genes when B. pertussis strains are grown under nonmodulating conditions. PMID- 8419299 TI - Sporulation gene spoIIB from Bacillus subtilis. AB - We have cloned and characterized the sporulation gene spoIIB from Bacillus subtilis. In extension of previous nucleotide sequence analysis, our results show that the order of genes in the vicinity of spoIIB is valS folC comC spoIIB orfA orfB mreB mreC mreD minC minD spoIVFA spoIVFB L20 orfX L24 spoOB obg pheB pheA. All 20 genes have the same orientation; the direction of transcription is from valS to pheA. We show that spoIIB is a 332-codon-long open reading frame whose transcription is under sporulation control. The deduced amino acid sequence of the spoIIB gene product, a 36-kDa polypeptide, is highly charged and contains a stretch of uncharged amino acids that could correspond to a transmembrane segment. Surprisingly, mutations in spoIIB, including an in vitro-constructed null mutation, cause only a mild impairment of spore formation in certain otherwise wild-type bacteria. However, when combined with mutations in another sporulation gene, spoVG, mutations in spoIIB cause a severe block in spore formation at the stage (stage II) of septum formation. (As with spoIIB mutations, mutations in spoVG cause little impairment in sporulation on their own.) The nature of the spoIIB spoVG mutant phenotype is discussed in terms of the events involved in the maturation of the sporulation septum and in the activation of sporulation transcription factors sigma F and sigma E. PMID- 8419301 TI - In vitro ferredoxin-dependent desaturation of fatty acids in cyanobacterial thylakoid membranes. AB - Thylakoid membranes isolated from the cyanobacterium Synechocystis sp. strain PCC6803 were capable of desaturating the acyl groups in monogalactosyl diacylglycerol. This desaturation reaction required the reduced form of ferredoxin. PMID- 8419300 TI - Nucleotide sequence of the Salmonella typhimurium mutB gene, the homolog of Escherichia coli mutY. AB - The mutB gene of Salmonella typhimurium is involved in a methylation-independent repair pathway specific for A/G or A/C mismatches and is the homolog of the Escherichia coli mutY gene. The mutB gene of S. typhimurium was cloned and sequenced. The isolated mutB clone reduced the mutation rate of the mutB mutant to wild-type levels and also restored A/G mismatch-specific nicking activity, which is defective in mutB extracts. The amino acid sequence encoded by the mutB gene is 91% homologous to that encoded by the E. coli mutY gene. PMID- 8419302 TI - Role of tol genes in cloacin DF13 susceptibility of Escherichia coli K-12 strains expressing the cloacin DF13-aerobactin receptor IutA. AB - IutA is the outer membrane protein receptor for ferric aerobactin and the bacteriocin cloacin DF13. Although the same receptor is shared, ferric aerobactin transport across the outer membrane in Escherichia coli is TonB dependent, whereas cloacin DF13 transport is not. We have recently observed that tolQ is required for cloacin DF13 susceptibility (J.A. Thomas and M.A. Valvano, FEMS Microbiol. Lett. 91:107-112, 1992). In this study, we demonstrate that the genes tolQ, tolR, and tolA, but not tolB, tolC, and ompF, are required for the internalization of cloacin DF13 and they are not involved in the transport of ferric aerobactin. PMID- 8419303 TI - Analysis of the topology of a membrane protein by using a minimum number of alkaline phosphatase fusions. AB - An approach to analyzing the topology of membrane proteins with alkaline phosphatase fusions is described. Precise fusions were constructed by using polymerase chain reaction at the C terminus of each hydrophilic region of the membrane protein. The disruption of topogenic signals is thereby minimized, and predictable anomalous results are avoided. The Escherichia coli MalG protein has been analyzed. PMID- 8419304 TI - The Helicobacter pylori 19.6-kilodalton protein is an iron-containing protein resembling ferritin. AB - The gastric pathogen Helicobacter pylori has been shown to produce a 19.6-kDa protein with apparent binding activity for erythrocytes, human buccal epithelial cells, and laminin. In this report we demonstrate that it is an iron-binding protein, resembling ferritin both structurally and biochemically. Also, because its binding activity for laminin, erythrocytes, and buccal cells was abolished by low concentrations of Tween 20, binding is likely nonspecific. PMID- 8419305 TI - Absence of a role for DNA polymerase II in SOS-induced translesion bypass of phi X174. AB - In order to examine the possible role of Escherichia coli DNA polymerase II in SOS-induced translesion bypass, Weigle reactivation and mutation induction were measured with single-stranded phi X174 transfecting DNA containing individual lesions. No decrease in bypass of thymine glycol or cyclobutane pyrimidine dimers in the absence of DNA polymerase II was observed. Furthermore, DNA polymerase II did not affect bypass of abasic sites when either survival or mutagenesis was the endpoint. Lastly, repair of gapped DNA molecules, intermediates in methyl directed mismatch repair, was also unaffected by the presence or absence of DNA polymerase II. PMID- 8419306 TI - Membrane topology model of Escherichia coli alpha-ketoglutarate permease by phoA fusion analysis. AB - Escherichia coli alpha-ketoglutarate permease (KgtP) is a 432-amino-acid protein that symports alpha-ketoglutarate and protons. KgtP was predicted to contain 12 membrane-spanning domains on the basis of a calculated hydropathy profile. The membrane topology model of KgtP was analyzed by using kgtP-phoA gene fusions and measuring alkaline phosphatase activities in cells expressing the chimeric proteins. Comparisons of the phosphatase activity levels and the locations of the KgtP-PhoA junctions are consistent with the predicted membrane topology model of KgtP. PMID- 8419307 TI - Characterization of the gcd gene from Escherichia coli K-12 W3110 and regulation of its expression. AB - DNA sequence and expressional analyses of the gcd gene of Escherichia coli K-12 W3110 revealed that two promoters that were detected were regulated negatively by cyclic AMP and positively by oxygen. Sequence conservation of the gcd gene between E. coli K-12 W3110 and PPA42 suggests that glucose dehydrogenase is required for the E. coli cells, even though it ordinarily exists as an apoprotein. PMID- 8419308 TI - Saliva-binding protein (SsaB) from Streptococcus sanguis 12 is a lipoprotein. AB - Two lipoprotein consensus sequences (Leu-X-X-Cys) are found in the presumptive signal peptide region (positions 12 to 15 and 17 to 20) of saliva-binding protein (SsaB) from Streptococcus sanguis 12. Three analogs of SsaB containing Cys-->Gly mutations were constructed by site-directed mutagenesis of pSA2, the recombinant plasmid expressing SsaB. [3H]palmitate was incorporated into SsaB only when the native Cys-20 residue was present. These data show that SsaB is a lipoprotein and that Cys-20 is the critical site for acylation. PMID- 8419309 TI - Modulation of the mitochondrial cyclosporin A-sensitive permeability transition pore. I. Evidence for two separate Me2+ binding sites with opposing effects on the pore open probability. AB - This paper reports an investigation on the regulation of the mitochondrial cyclosporin A-sensitive permeability transition pore (MTP). Energized, coupled rat liver mitochondria incubated in sucrose medium in the presence of phosphate maintain a high proton electrochemical gradient (delta microH) and a low permeability to solutes. Addition of a small (10-20 microM) Ca2+ pulse leads to a transient membrane depolarization. After Ca2+ accumulation, a high delta microH is recovered, and mitochondria remain coupled indefinitely. Yet, addition of fully uncoupling concentrations of carbonyl cyanide-p-trifluoromethoxyphenyl hydrazone (FCCP) brings about MTP opening within seconds. This finding confirms that MTP opening is the consequence rather than the cause of membrane depolarization, and allowed us to study the operation of the MTP in a synchronized population of mitochondria, since pore opening can be triggered by the addition of uncoupler under a series of experimental conditions. We find that three regulatory sites can be defined: (i) an internal Me2+ binding site: when this site is occupied by Ca2+, the pore "open" probability increases, while other Me2+ ions (Sr2+, Mn2+) have an inhibitory effect; (ii) an external Me2+ binding site: when this site is occupied by Me2+ ions, including Ca2+, the pore open probability decreases; (iii) an independent cyclosporin A binding site: when this site is occupied by cyclosporin A the pore open probability decreases. We show that at variance from the case of cyclosporin A, MTP inhibition by the phospholipase A2 inhibitors nupercaine and trifluoperazine is Ca(2+)-competitive and is presumably related to interference by these drugs with Ca2+ binding to the internal regulatory site. PMID- 8419310 TI - 5,10-Dideazatetrahydrofolic acid (DDATHF) transport in CCRF-CEM and MA104 cell lines. AB - 5,10-Dideazatetrahydrolic acid (DDATHF) is representative of a new class of antifolates acting through inhibition of de novo purine synthesis. We report here the transport characteristics of the diastereomers of DDATHF, which differ in configuration at C6, and comparison studies with other folate and antifolate analogs. (6R)-DDATHF showed high affinity for the influx system of CCRF-CEM cells with a Km of 1.07 microM and an influx Vmax of 4.04 pmol/min/10(7) cells. Comparative studies with methotrexate yielded an influx Km of 4.98 microM and a Vmax of 6.64 pmol/min/10(7) cells, and with 5-formyltetrahydrofolate an influx Km of 2.18 microM and a Vmax of 6.84 pmol/min/10(7) cells. Uptake of (6R)-DDATHF was competitively inhibited by (6S)-DDATHF, methotrexate (MTX), and 5 formyltetrahydrofolate, all with Ki values similar to their influx Km. The (6S) DDATHF diastereomer had an influx Km of 1.04 microM, similar to that of (6R) DDATHF; however, the Vmax of 1.72 pmol/min/10(7) cells was 2.3-fold lower than for (6R)-DDATHF. The transport properties of DDATHF were also studied in a mutant cell line (CEM/MTX), resistant to MTX based on impaired drug transport. In this system (6R)-DDATHF showed an influx Km of 1.49 microM and a decreased influx Vmax of 0.60 pmol/min/10(7) cells. A similar effect was shown for MTX (Km of 7.48 microM, Vmax of 1.02 pmol/min/10(7) cells). The number of binding sites in CCRF CEM cells was similar for (6R)-DDATHF, (6S)-DDATHF, and MTX, 0.74, 0.71, and 0.76 pmol/10(7) cells, respectively. These values were slightly higher in the CEM/MTX cell line (1.07 and 1.09 pmol/10(7) cells for (6R)-DDATHF and MTX, respectively). Treatment of CCRF-CEM cells with either the N-hydroxysuccinimide ester of MTX or the corresponding N-hydroxysuccinimide ester of (6R)-DDATHF caused substantial inhibition (> 90%) of the influx of (6R)-[3H]DDATHF and [3H]MTX, respectively. These results suggest strongly that DDATHF and MTX share a common influx mechanism through the reduced folate transport system. The internalization of DDATHF by monkey kidney epithelial MA104 cells, which express a high affinity folate receptor, was also studied. Competitive binding studies using purified folate receptor and radiolabeled 5-methyltetrahydrofolate showed that (6S)- and (6R)-DDATHF both had I50 values lower than 5-methyltetrahydrofolate (12 nM). Further studies indicate that both DDATHF isomers are actively intracellularly concentrated through this route and are also rapidly converted to high chain length polyglutamates. Transport via this system was inhibited in folate-depleted cells by 10 nM folic acid.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419311 TI - Characterization of metal ion-induced [3H]inositol hexakisphosphate binding to rat cerebellar membranes. AB - The binding of [3H]inositol hexakisphosphate ([3H] InsP6) to rat cerebellar membranes has been characterized with the objective of establishing the role, if any, of a membrane protein receptor. In the presence of EDTA, we have previously identified an InsP6-binding site with a capacity of approximately 20 pmol/mg protein (Hawkins, P. T., Reynolds, D. J. M., Poyner, D. R., and Hanley, M. R. (1990) Biochem. Biophys. Res. Commun. 167, 819-827). However, in the presence of 1 mM Mg2+, the capacity of [3H]InsP6 binding to membranes was increased approximately 9-fold. This enhancing effect of Mg2+ was reversed by addition of 10 microM of several cation chelators, suggesting that the increased binding required trace quantities of other metal cations. This is supported by experiments where it was possible to saturate binding by addition of excess membranes, despite not significantly depleting radioligand, pointing to removal of some other factor. Removal of endogenous cations from the binding assay by pretreatment with chelex resin also prevents the Mg(2+)-induced potentiation. Consideration of the specificity of the chelators able to abolish this potentiation suggested involvement of Fe3+ or Al3+. Both these ions (but not several others) were able to increase [3H]InsP6 binding to chelex-pretreated membranes at concentrations of 1 microM. It is possible to demonstrate synergy between Fe3+ and Mg2+ under these conditions. We propose that [3H]InsP6 may interact with membranes through non-protein recognition, possibly via phospholipids, in a manner dependent upon trace metals. The implications of this for InsP6 biology are considered. PMID- 8419312 TI - Cloning, genomic organization, and chromosomal localization of human cathepsin L. AB - Cathepsin L is a lysosomal cysteine protease whose expression and secretion is induced by malignant transformation, growth factors, and tumor promoters. Many human tumors express high levels of cathepsin L, which is a broad spectrum protease with potent elastase and collagenase activities. Two published human cathepsin L cDNA sequences differ only in their 5'-untranslated regions. In this study, we demonstrate the concurrent expression of two distinct human cathepsin L mRNAs (hCATL-A and hCATL-B) in adenocarcinoma, hepatoma, and renal cancer cell lines. Cloning of the human cathepsin L gene by polymerase chain reaction amplification of genomic DNA and subsequent sequencing reveals that hCATL-A and hCATL-B mRNAs are encoded by a single gene. The 3' end of the first intron contains the 5' portion of hCATL-B and is contiguous to the second exon of the gene. These data suggest either the possibility of alternative splicing or the presence of a second promoter within the first intron of the hCATL gene. We mapped the hCATL gene to chromosome 9q21-22. Sequencing of both the mouse and human cathepsin L genes demonstrates almost complete conservation of exon and intron position, but significant divergence in intron structure, possibly reflecting differences in regulation of expression of the mouse and human cathepsin L genes. PMID- 8419313 TI - Inducer-dependent transcriptional activation of the P4501A2 gene in vivo and in isolated hepatocytes. AB - In vitro nuclear run-on transcription analysis using probes directed against different regions of CYP1A2 revealed that the 70-100-fold induction of CYP1A2 mRNA by polycyclic aromatic compounds is associated with a corresponding increase in the transcriptional activation of this gene in rat liver. Probes from regions of the 1st, 2nd, and 4th introns detected approximately 50-100-fold higher CYP1A2 run-on transcription in liver nuclei from inducer-treated animals than in nuclei from untreated animals. The run-on signals from untreated rats were 3-5-fold above background signals. Additional experiments using single-stranded DNA probes and a probe from a region 5' to the CYP1A2 transcription start site revealed that the inducer-dependent transcripts were colinear with the CYP1A2 mRNA and that they did not result from read through of an initiation event 5' to CYP1A2. Run-on transcription analyses were also carried out with nuclei from isolated hepatocytes using the same series of probes spanning CYP1A2. These analyses indicated that the inducer-dependent accumulation of CYP1A2 mRNA in hepatocytes is associated with at least a 20-fold increase in CYP1A2 transcription. In contrast to liver and hepatocytes, these probes failed to detect run-on transcripts from kidney nuclei, indicating that the lack of CYP1A2 mRNA in this tissue is due to the lack of transcriptional activation of this gene by polycyclic aromatic compounds. PMID- 8419314 TI - Characterization of the integrin specificities of disintegrins isolated from American pit viper venoms. AB - A new series of homologous disintegrins was isolated from the venoms of new world pit viper genus Bothrops, Crotalus, and Lachesis. The relative activities of each disintegrin in blocking adhesive protein binding activities of GPIIb-IIIa, alpha v beta 3, and alpha 5 beta 1 were determined and correlated with their primary amino acid sequences. Four disintegrins contained the RGDW sequence and were found to be approximately twice as effective in blocking the binding of fibrinogen to GPIIb-IIIa than inhibiting the binding of vitronectin to alpha v beta 3 in solid-phase ligand binding assays (IC50 = 7.3 and 17.2 nM, respectively). A second group of seven disintegrins contained the RGDNP sequence and were found to be more potent inhibitors of vitronectin binding to alpha v beta 3 than fibrinogen binding to GPIIb-IIIa (IC50 = 4.3 and 19 nM, respectively). The RGDNP containing disintegrins were also greater than 10-fold more potent than RGDW containing disintegrins in blocking the adhesion of cells mediated by alpha 5 beta 1. These data illustrate that amino acid sequences immediately adjacent to the RGD site of disintegrins can create an extended RGD locus which coupled with conformational display of the RGD sequence may be involved in determining integrin selectivity and affinity. This information has been used in separate studies to design conformationally constrained integrin antagonists with high affinity for platelet GPIIb-IIIa. PMID- 8419315 TI - Design of potent and specific integrin antagonists. Peptide antagonists with high specificity for glycoprotein IIb-IIIa. AB - Members of the snake venon-derived, "disintegrin" peptide family containing the Arg-Gly-Asp (RGD) amino acid sequence are among the most potent inhibitors of the binding of adhesive proteins to platelet glycoprotein (GP) IIb-IIIa. However, GPIIb-IIIa antagonists containing the RGD sequence are not integrin specific and inhibit the adhesive functions of many other RGD-dependent integrins. The single disintegrin peptide, barbourin, containing a conservative amino acid substitution of Lys (K) for Arg (R) in the RGD sequence, is however, highly specific for GPIIb IIIa. Using this information we have tested the hypothesis that both structural and conformational elements of barbourin are important for its high affinity and selectivity for platelet GPIIb-IIIa by synthesizing a series of conformationally constrained, disulfide-bridged peptides containing the KGD amino acid sequence. Incorporation of the KGD sequence into a cyclic peptide template, followed by systematic optimization of the cyclic ring size, optimization of secondary hydrophobic binding site interactions, and the derivatization of the lysyl side chain functionality of the KGD sequence has resulted in peptide analogs which display inhibitory potency and GPIIb-IIIa selectivity comparable to that of barbourin. This study demonstrates that the specificity and potency of the disintegrin family of antagonists, in particular barbourin, can be mimicked by small, conformationally restrained peptides. PMID- 8419316 TI - Site-directed mutagenesis identifies aspartate 33 as a previously unidentified critical residue in the catalytic mechanism of rabbit aldolase A. AB - The expression and purification of the rabbit muscle aldolase A (D-fructose 1,6 bisphosphate:D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13) from an expression plasmid in bacteria is described. The enzyme is produced in bacteria at a level of 300 mg/liter and is indistinguishable from the enzyme isolated from muscle in assays using fructose 1,6-bisphosphate and fructose 1-phosphate. The recombinant enzyme has the same primary, secondary, and quaternary structure as the muscle enzyme. Aspartic acid 33, found near the active site lysine in the crystal structure, is changed to alanine, serine, and glutamic acid by site-directed mutagenesis, resulting in the mutant proteins, D33A, D33S, and D33E, respectively. The mutant enzymes are purified by substrate affinity elution from carboxylmethyl-Sepharose, the same method as that used for the wild-type enzyme. The secondary and quaternary structure of D33A is identical to wild-type aldolase when analyzed by light scattering, gel filtration, and circular dichroism. Moreover, the hexose substrate can be fixed in the active site by reduction of the Schiff base with sodium borohydride, indicating that the active site is not drastically altered. These single mutations in the active site have a serious effect on the activity of the enzyme. In addition, the rate of carbanion oxidation for D33A is 17-29 times slower when the substrate is fructose 1,6 bisphosphate versus dihydroxyacetone phosphate, whereas in the wild-type there is no significant difference in these rates. This evidence and the conservation of this residue in other class I aldolases indicate that aspartic acid 33 is an essential residue in the catalytic mechanism, possibly involved in abstraction of the carbon 4 hydroxyl proton. PMID- 8419317 TI - Mutations of the FLP recombinase gene that cause a deficiency in DNA bending and strand cleavage. AB - We have used site-directed mutagenesis to change several amino acids of the C terminal portion of the FLP recombinase of Saccharomyces cerevisiae. These residues are absolutely conserved among the six FLP-like proteins from various yeast strains. We have examined the ability of the altered proteins to catalyze recombination in vivo and in vitro and to perform various partial steps of the reaction in vitro. Two of the mutations produced a partial defect in DNA binding but the remainder resulted in normal binding. All of these mutations caused impairment of the ability of the protein to induce the type II bend of the FRT site, and some of these proteins were also defective in DNA strand cleavage. None of the mutations affected the ability of the proteins to perform synapsis between two FRT sites, but some were defective in strand ligation. Interestingly, some mutant proteins showed impairment of the initial stages of the recombination reaction on a linear substrate and yet they maintained the ability to resolve a Holliday intermediate in the reaction. We conclude that this conserved region of the FLP protein is important for the early stage(s) of the recombination reaction. PMID- 8419318 TI - Nucleosome structural changes during derepression of silent mating-type loci in yeast. AB - Mutant a and alpha yeast cells were created with histone H3 containing cysteine in place of alanine 110. Because transcriptionally active nucleosomes "unfold" to reveal the histone H3-thiol groups at the center of the core, the active nucleosomes of the mutant strain can be isolated by mercury-affinity chromatography. We compared the unbound and mercury-bound nucleosomes of haploid H3-mutant strains expressing either the MAT alpha or the MATa mating-type locus. In a MAT alpha strain, the Hg-bound nucleosomes are enriched in MAT alpha DNA but lack the DNA of the transcriptionally silent HMRa mating-type locus. Conversely, in a MATa strain, the Hg-bound nucleosomes are enriched in MATa DNA sequences but deficient in HML alpha DNA. When the SIR3 gene, known to be required for silencing of the repressed mating-type loci, is mutated in the MAT alpha strain, transcription of the HMRa ensues, and its nucleosomes, as well as those of the MAT alpha locus, are retained by the organomercurial column. It follows that derepression of the silent mating-type locus, caused by the sir3 null mutation, is accompanied by an unfolding of its nucleosomes to reveal the histone H3 SH groups at their centers. Nucleosomes of the pheromone-encoding gene MFA2, a gene that is expressed in MATa cells but not in MAT alpha cells, are bound to the organomercurial column when isolated from MATa cells but not from MAT alpha cells. Thus, there is a good correlation between nucleosome unfolding and the renewed transcriptional activity at mating-type loci, and at MFA2, which had been silenced for prolonged periods. A close temporal correlation between nucleosome refolding and the cessation of transcription is not always observed in yeast, however, in contrast to observations in mammalian cells. For example, nucleosomes of the GAL1 gene are maintained in a "poised" or "primed" thiol-reactive state even when the gene is not being transcribed (Chen, T. A., Smith, M. M., Le, S., Sternglanz, R., and Allfrey, V. G. (1991) J. Biol. Chem. 266, 6489-6498). It follows that the unfolding of the nucleosome cores of the yeast H3 mutant is regulated by factors that are not temporally linked to the recruitment or traverse of the RNA polymerase complex, but which may determine the rate at which different domains of chromatin adapt to the need for transcription of the associated DNA sequences. PMID- 8419319 TI - Identification and purification of a self-associating copper-binding protein from mouse hepatic cytosols. AB - The proteins which bind copper when it first enters cells and deliver that copper to the cellular sites where copper is incorporated into copper enzymes are unknown. When radiolabeled (64Cu(II)) cytosol proteins were fractionated on Superose columns, two large copper-binding protein fractions were detected which together bound 63% of the total 64Cu recovered from the column. A self associating, 50-kDa copper-binding protein was identified whose tetramer and dimer appeared to elute in one of the major copper-binding fractions and the monomer eluted in the other. This protein was purified to homogeneity by successive Mono Q, chelating Superose, and phenyl-Superose columns. The concentrated, purified protein showed high amounts of tetramer and monomer plus some dimer, each of which bound copper. Rechromatography of the tetramer-dimer fraction on Superose gave rise to approximately equal amounts of tetramer, dimer, and monomer. Similarly, rechromatography of the monomer fraction gave rise to dimer and tetramer. Thus, the results with the purified protein confirmed that the 50-kDa protein exists as a tetramer-dimer-monomer equilibrium. The 50-kDa protein apparently makes a major contribution to the total copper-binding activity of the mouse hepatic cytosol and may play a significant role in intracellular copper metabolism. PMID- 8419320 TI - Purification and properties of a self-associating, 50-kDa copper-binding protein from brindled mouse livers. AB - The brindled mouse is an animal model of Menkes disease, a fatal, X-linked disease of copper metabolism. A self-associating, 50-kDa copper-binding protein (CuBP) was purified from brindled mouse hepatic cytosols, and some of its properties were determined. When 64Cu-labeled whole hepatic cytosols were fractionated on Superose, statistically significantly less than normal 64Cu binding was detected in both the fraction which contained the tetramer plus dimer (approximately 26% less) and the fraction containing the monomer of CuBP (approximately 37% less). CuBP was purified from brindled mouse hepatic cytosols by successive Mono Q, chelating Superose, and phenyl-Superose columns using the same methods used to purify the protein from normal mice. However, CuBP from the brindled mice was somewhat unstable during the purification. Also, CuBP from the brindled mouse eluted abnormally from the phenyl-Superose column. Thus, while the protein from normal mice eluted at approximately 20 min after starting the final water elution step, the brindled mouse protein eluted by approximately 5 min. This seemed to be due to abnormal self-association in the column buffers. Consistent with the results using whole cytosols, the purified CuBP from the brindled mouse showed decreased copper binding in both the tetramer and monomer fractions from Superose. Moreover, under the same conditions, CuBP from the brindled mice seemed to have relatively less tetramer and more dimer than normal. The results are consistent with a significant role for CuBP in intracellular copper metabolism, and an abnormal structure of CuBP may be the basic defect in the brindled mice and, by inference, Menkes disease. PMID- 8419321 TI - Phosphorylation of recombinant tau by cAMP-dependent protein kinase. Identification of phosphorylation sites and effect on microtubule assembly. AB - Tau protein is an integral component of paired helical filaments, a pathological feature of Alzheimer's disease. tau extracted from these filaments displays decreased electrophoretic mobility due to aberrant phosphorylation. Here we show that recombinant human tau can be phosphorylated by cAMP-dependent protein kinase resulting in decreased electrophoretic mobility. Phosphorylation of tau by cAMP dependent protein kinase caused a 92% decrease in the maximum rate of tau-induced microtubule assembly. The sites of phosphorylation were identified by digesting phosphorylated tau with proteases, separating the peptides by reversed-phase HPLC, and analyzing the isolated peptides by liquid-secondary ion mass spectrometry and solid-phase N-terminal sequencing. Five phosphorylation sites were identified, two of which were located within microtubule binding domains. One site was previously shown to be the sole phosphorylation site for CaM kinase II; phosphorylation at this site by CaM kinase II was sufficient to cause decreased electrophoretic mobility (Steiner, B., Mandelkow, E. M., Biernat, J., Gustke, N., Meyer, H. E., Schmidt, B., Mieskes, G., Soling, H. D., Drechsel, D., Kirschner, M. W., Goedert, M., and Mandelkow, E. (1990) EMBO J. 9, 3539-3544). Thus two different second messenger-dependent protein kinases can phosphorylate tau at the same site and induce a shift in tau mobility like that seen in Alzheimer's disease. PMID- 8419322 TI - Purification and characterization of human lysosomal protective protein expressed in stably transformed Chinese hamster ovary cells. AB - Chinese hamster ovary cells were transfected with a recombinant DNA containing the entire coding sequence of human lysosomal protective protein cDNA under the control of mouse metallothionein I promoter. Neomycin and methotrexate-resistant stably transformed cell lines expressing this protein were isolated. Immunoprecipitation of the product with antiserum against human placental protective protein-beta-galactosidase complex revealed a 52-kDa protective protein precursor, which was then processed to mature form, a heterodimer of 32- and 20-kDa polypeptides. The precursor secreted in the culture medium was taken up by the mannose 6-phosphate receptor system and restored acid carboxypeptidase, beta-galactosidase, and neuraminidase activities in galactosialidosis fibroblasts. The expressed protein showed a granular pattern in intracellular distribution, was fractionated at the density of lysosomes, and had serine esterase activities; acid carboxypeptidase at pH 5.6, esterase at pH 7.0, and carboxyl-terminal deamidase at pH 7.0. They were inhibited simultaneously by phenylmethylsulfonyl fluoride, N-benzyloxycarbonyl-L-phenylalanine chloromethyl ketone, or iodoacetamide. The acid carboxypeptidase activity of the purified monomeric mature protective protein was labile in vitro under the acidic condition. Saposins (sphingolipid activator proteins) stabilized the activity at micromolar level concentrations. PMID- 8419323 TI - Characterization of the gene for rat phosphorylase kinase catalytic subunit. AB - Phosphorylase kinase, a key enzyme in glycogen metabolism, has a subunit composition of (alpha beta gamma delta)4, in which the alpha and beta subunits are regulatory, delta is calmodulin, and the gamma subunit is catalytic. As one segment of our studies on the regulation of the expression of phosphorylase kinase subunits, we present in this report the structure of the gene for the catalytic gamma subunit. The gene extends over 16 kilobase pairs (kb) of DNA, and contains eight introns within the coding region plus one 3.3-kb intron upstream in the 5'-untranslated region. Within this first intron, and also upstream of the transcription start site, are sequences homologous to defined regulatory elements, including some found in other muscle-specific genes. The positions of intron splice junctions for this gene have been compared with similar data for other protein kinase genes. A somewhat unexpected finding for the gamma subunit is that two of the splice junctions fall in the midst of highly conserved strings of amino acids, both of which have been nominally defined as functional domains for the protein kinases and appear to make key contributions to substrate binding and phosphotransferase catalysis. PMID- 8419324 TI - Intracellular degradation of Fc gamma RIII in mouse bone marrow culture-derived progenitor mast cells prevents its surface expression and associated function. AB - Although mouse interleukin-3-dependent, bone marrow culture-derived progenitor mast cells (BMMC) and a Kirsten sarcoma virus (KiSV)-immortalized mouse mast cell line (MC4w) both express on their surfaces receptors for the Fc portion of IgG (Fc gamma R), only MC4w degranulate upon Fc gamma R perturbation. As shown by surface iodination and SDS-polyacrylamide gel electrophoresis analysis of deglycosylated proteins immunoprecipitated with the Fc gamma R-specific monoclonal antibody 2.4G2, a 26-kDa protein, identified as Fc gamma RIII by immunoblotting with antibody to Fc gamma RIII, was predominantly expressed on the surface of MC4w but minimally on BMMC. However, both BMMC and MC4w expressed mRNA for Fc gamma RIII as determined by RNA blot analysis, and both translated Fc gamma RIII as assessed by intrinsic radiolabeling and SDS-polyacrylamide gel electrophoresis analysis of deglycosylated monoclonal antibody 2.4G2 immunoprecipitates. Pulse-chase analysis showed that intrinsically radiolabeled Fc gamma RIII was stable in MC4w cells but was degraded rapidly in BMMC and that newly synthesized Fc gamma RIII remained sensitive to digestion by endoglycosidase H in BMMC but rapidly became resistant in MC4w. These data suggest that the deficiency in surface Fc gamma RIII expression on BMMC is due to the degradation of Fc gamma RIII in the endoplasmic reticulum. Immunoprecipitation of surface Fc gamma R and Fc receptors for IgE (Fc epsilon RI) from digitonin-extracted cells followed by immunoblotting with antibody to Fc epsilon RI gamma-chain showed that gamma-chain is associated with surface Fc epsilon RI and Fc gamma R in MC4w, but only with Fc epsilon RI in BMMC, which lack surface Fc gamma RIII. Inasmuch as BMMC are progenitors of serosal mast cells, which, like MC4w, express surface Fc gamma RIII and undergo Fc gamma R mediated activation, the data suggest that maturation of BMMC enables Fc gamma RIII to bypass degradation in the endoplasmic reticulum, resulting in the acquisition of functional Fc gamma RIII/gamma-chain complexes on the cell surface. PMID- 8419325 TI - Rod structure of a phycoerythrin II-containing phycobilisome. I. Organization and sequence of the gene cluster encoding the major phycobiliprotein rod components in the genome of marine Synechococcus sp. WH8020. AB - Phycobilisomes of the unicellular marine cyanobacteria are unique in having rod substructures with two distinct phycoerythrins, PE I and PE II, with five and six bilins, respectively (Ong, L. J., and Glazer, A. N. (1991) J. Biol. Chem. 266, 9515-9527). The genes for the alpha and beta subunits of PE I, PE II, and phycocyanin, and that for the PE II-associated linker polypeptide, are clustered on a single 15-kilobase region of the genome of Synechococcus sp. WH8020. Complete sequencing of this region allowed definitive assignment of the positions of all bilin attachment sites in these phycobiliproteins. Twelve other open reading frames are closely associated with the structural genes specified above. Six are homologous to open reading frames adjacent to phycobiliprotein genes in other cyanobacteria and inferred to be involved in bilin addition. This is the largest number of open reading frames of this class known in any cyanobacterium. Another of the open reading frames has a short region of striking similarity to the active site sequence of a bovine protein-phosphotyrosine phosphatase. PMID- 8419326 TI - Rod structure of a phycoerythrin II-containing phycobilisome. II. Complete sequence and bilin attachment site of a phycoerythrin gamma subunit. AB - The major phycoerythrins of marine unicellular cyanobacteria Synechococcus spp. and those of red algae are isolated as stable complexes with the composition (alpha beta)6 gamma. The gamma subunits carry bilins and in this respect are unique among phycobilisome rod linker polypeptides. There has been no complete amino acid sequence data on any gamma subunit. Synechococcus sp. WH8020 phycoerythrin II (PE II) gamma subunit was isolated from purified PE II hexamers. Partial amino acid sequence determination showed that the gamma subunit was encoded by the mpeC gene, an open reading frame 275 base pairs 3' of mpeA and mpeB, which encode the alpha and beta subunits of PE II, respectively (Wilbanks, S. M., and Glazer, A. N. (1993) J. Biol. Chem. 268, 1226-1235). A single phycourobilin is attached through a thioether bond to gamma-Cys-49. Derivatization with 4-vinylpyridine showed that the only other cysteinyl residue, gamma-Cys-64, is unsubstituted. MpeC encodes a polypeptide of 293 residues with a predicted molecular weight of 32,100 and a pI of 8.9. These properties are like those of non-bilin-bearing linker polypeptides associated with C-phycoerythrin and hence the gamma subunit is designated gamma (LR32). Alignment of the sequence of the PE II-gamma with those of the latter polypeptides shows that PE II-gamma has a 49-residue extension at the N terminus, that encompasses the phycourobilin attachment site, and is shorter by a similar number of residues at the C terminus. These differences in linker polypeptide structure offer a possible explanation for the observed much higher stability of PE II hexamers relative to those of C-phycoerythrins. PMID- 8419327 TI - Branching beta 1-6N-acetylglucosaminetransferases and polylactosamine expression in mouse F9 teratocarcinoma cells and differentiated counterparts. AB - beta-All-trans-retinoic acid (RA)-induced endodermal differentiation of mouse F9 teratocarcinoma cells is accompanied by changes in glycoprotein glycosylation, including expression of i antigen (i.e. polylactosamine) and leukophytohemagglutinin-reactive oligosaccharides (i.e. -GlcNAc beta 1-6Man alpha 1-6-branched N-linked). We have used the F9 teratocarcinoma cells as a model to study developmental regulation of glycosyltransferase activities which are responsible for the biosynthesis of beta 1-6GlcNAc-branched N- and O-linked oligosaccharides and polylactosamine. Growth of F9 cells in the presence of 10( 6) M RA for 4 days increased core 2 GlcNAc transferase and GlcNAc transferase V activities by 13- and 6-fold, respectively, whereas the activities of GlcNAc transferase I, beta 1-3GlcNAc transferase (i), beta 1-4Gal transferase, and beta 1-3Gal transferase increased 2-4-fold. Induction of glycosyltransferase activities by RA was dose-dependent and showed a biphasic response with approximately half of the increase observed 3 days after RA treatment and the remainder occurred by day 4. PYS-2, a parietal endoderm cell line, showed levels of glycosyltransferase activities similar to those of RA-treated F9 cells. Glycosyltransferase activities in the RA-resistant F9 cell line (RA-3-10) were low and showed only a small induction by RA. These observations suggest that differentiation of F9 cells is closely associated with induction of multiple glycosyltransferase activities, with most pronounced increases in GlcNAc transferase V and 2',5'-tetradenylate (core 2) GlcNAc transferase. The increase in GlcNAc transferase V was also reflected by the 4-6-fold increase in the binding of 125I-leukophytohemagglutinin to several cellular glycoproteins, which occurred after 3 days of RA treatment. The endo-beta-galactosidase-sensitive polylactosamine content of membrane glycoproteins and, in particular, the LAMP-1 glycoprotein was markedly increased after RA treatment of F9 cells. Consistent with these observations, fucosylated polylactosamine (i.e. dimeric Lex) was also increased in RA-treated cells. Analysis of the aryl oligosaccharides produced by F9 cells cultured in the presence of aryl alpha-D-GalNAc showed that RA treatment enhanced the synthesis of disialyl core 2 O-linked oligosaccharides and increased the polylactosamine content of the aryl oligosaccharides by > 20-fold. The results suggest that differentiation of F9 cells into endoderm is closely associated with increased GlcNAc transferase V and core 2 GlcNAc transferase activities, enzymes which control the level of beta 1-6GlcNAc-branched N- and O linked oligosaccharides, the preferred substrates for polylactosamine addition. PMID- 8419328 TI - Purification and characterization of the SRS2 DNA helicase of the yeast Saccharomyces cerevisiae. AB - The SRS2 gene of Saccharomyces cerevisiae was identified through mutational analysis as a suppressor of radiation-sensitive mutations in the error-prone repair pathway and by a hyper-recombination phenotype. Comparison of the derived amino acid sequence revealed the gene to have high homology to the bacterial DNA helicases UvrD and Rep (Aboussekhra, A., Chanet, R., Zgaga, Z., Cassier-Chauvat, C., Heude, M., and Fabre, F. (1989) Nucleic Acids Res. 17, 7211-7219). We have purified the SRS2 protein from Escherichia coli extracts by tagging the SRS2 gene with 6 carboxyl-terminal histidine residues and overexpressing the tagged protein in a pET-3c vector. Extracts were passed over a metal-chelating affinity chromatography column followed by gel filtration to obtain an enriched protein fraction. Sephacryl gel filtration of pooled fractions containing the SRS2 protein yielded purified SRS2 protein by Coomassie Blue stain of SDS polyacrylamide gel electrophoresis gels. The purified SRS2 protein was found to have in vitro DNA-dependent ATPase and DNA helicase activities. The polarity of the helicase activity was determined to be 3' to 5', the same polarity as that found for the UvrD and Rep proteins. The carboxyl-terminal region of the protein is shown to contain a sequence for nuclear localization. Expression of the SRS2 in yeast was examined and found to be extremely low. PMID- 8419329 TI - The regulation of thymidine kinase in HL-60 human promyeloleukemia cells. AB - It has been well established that the regulation of thymidine kinase (TK) expression is highly growth-dependent. In this report, we present evidence that TK expression in undifferentiated HL-60 cells is not stringently controlled in a growth-dependent manner, except for a very moderate activation of TK in response to growth stimulation. Moreover, we have demonstrated for the first time that TK becomes phosphorylated, and the fluctuation of TK activity in these cells is related to the extent of phosphorylation of seryl residues of the TK polypeptide. This is further reinforced by the observation that the presence of Ser/Thr phosphatases inhibitor in the crude extract increases TK activity. Our data suggest that post-translational modification by phosphorylation is implicated in TK regulation in HL-60 cells. PMID- 8419330 TI - Hormonal regulation and properties of a new group of basic hemolymph proteins expressed during insect metamorphosis. AB - Two abundant basic proteins, and arylphorin, that are expressed during metamorphosis of Trichoplusia ni were isolated, their cDNAs cloned, and regulation of their expression assessed. According to the criteria of encoded protein sequence, positions of sequence coinitiation and cotermination, amino acid compositions, nonsex-specificity, and immunological analyses, the related yet distinct basic proteins do not correspond to any previously established group of insect proteins, each of which can be distinguished on the basis of such criteria, although they appear to be within the arthropod hemocyanin superfamily. The basic proteins can be separated under native isoelectric focusing conditions, indicating that they do not preferentially form heteromeric complexes under such conditions. While the transcripts for both basic proteins and a coexpressed acidic protein (Jones, G., Brown, N., Manczak, M., Hiremath, S., and Kafatos, F. (1990) J. Biol. Chem. 265, 8596-8602) appeared in both sexes on the day prior to ecdysteroid-driven metamorphic commitment, and persisted at high abundance for the next 24 h, the arylphorin mRNA appeared 1 day earlier and declined immediately after metamorphic commitment. The dynamics of suppression of the appearance and abundance of transcripts in response to maintenance of a high juvenile hormone titer was different for the acidic versus the two basic proteins. Another regulatory difference in response to juvenile hormone was shown by a decreased translatability of the mRNAs for the two basic proteins, but not for the acidic protein. In contrast, the abundance and translatability of arylphorin mRNA was insensitive to a high juvenile hormone level. A further difference in regulation was shown by the maintenance of a high level of both arylphorin and the acidic protein in the hemolymph after disappearance of their transcripts, while the basic proteins instead disappeared from the hemolymph and persisted in the fat body. These results establish at least three qualitatively different effects of juvenile hormone on the abundance of transcripts for these representatives of three groups in the hemocyanin superfamily, and also establish mRNA translatability as another differential level of regulation among them. Given these differences in mechanisms of regulation of evolutionarily related proteins, this system of metamorphosis-associated proteins in T. ni should prove to be a valuable tool in studies of mechanisms of hormonal regulation of gene expression during the metamorphic transformation of larval tissues. PMID- 8419331 TI - Alteration of the nucleoside triphosphate (NTP) catalytic domain within Escherichia coli recA protein attenuates NTP hydrolysis but not joint molecule formation. AB - The hydrolysis of the nucleoside triphosphates, such as ATP or GTP, plays a central role in a variety of biochemical processes; but, in most cases, the specific mechanism of energy transduction is unclear. DNA strand exchange promoted by the Escherichia coli recA protein is normally associated with ATP hydrolysis. However, we advanced the idea that the observed ATP hydrolysis is not obligatorily linked to the exchange of DNA strands (Menetski, J. P., Bear, D. G., and Kowalczykowski, S. C. (1990) Proc. Natl. Acad. Sci. U. S. A. 87, 21-25); instead, ATP binding resulting in an allosteric transition to an active form of the recA protein is sufficient. In this paper, we extend this conclusion by introducing a mutation within a highly conserved region of the recA protein that, on the basis of sequence similarity, is proposed to interact with the pyrophosphate moiety of a bound NTP molecule. The conservative substitution of an arginine for the invariant lysine at position 72 reduces NTP hydrolysis by approximately 600-850-fold. This mutation does not significantly alter the capacity of the mutant recA (K72R) protein either to bind nucleotide cofactors and single-stranded DNA or to respond allosterically to nucleotide cofactor binding. Despite the dramatic attenuation in NTP hydrolysis, the recA (K72R) protein retains the ability to promote homologous pairing and extensive exchange of DNA strands (up to 1.5 kilobase pairs). These results both identify a component of the catalytic domain for NTP hydrolysis and demonstrate that the recA protein-promoted pairing and exchange of DNA strands mechanistically require the allosteric transition induced by NTP cofactor binding, but not the energy educed from NTP hydrolysis. PMID- 8419332 TI - Ascorbic acid oxidation product(s) protect human low density lipoprotein against atherogenic modification. Anti- rather than prooxidant activity of vitamin C in the presence of transition metal ions. AB - The oxidative modification of low density lipoprotein (LDL) has been proposed as an important causative event in the development of human atherosclerosis. As a corollary of this hypothesis, antioxidants that can prevent LDL oxidation may inhibit atherosclerosis. Oxidative modification of LDL in vitro, either induced by Cu2+ or mediated by cultured arterial wall cells in media containing trace amounts of transition metal ions, is strongly inhibited by vitamin C (L-ascorbic acid (AA)). AA, however, is known to act as a prooxidant rather than an antioxidant in the presence of transition metal ions. We observed that AA is oxidized rapidly when incubated with Cu2+ and LDL, leading to transient formation of dehydro-L-ascorbic acid (DHA). Although AA and DHA can no longer be detected after 3.5 h of incubation, LDL resists oxidative modification for at least 20 h, as assessed by anodic gel electrophoretic mobility. Remarkably, DHA protects LDL more effectively against both Cu(2+)-induced lipid peroxidation and shifts in electrophoretic mobility than does AA; indeed, AA per se, without oxidation to DHA, offers no protection. By inhibiting oxidative modification of LDL, AA and DHA prevent uptake of LDL by macrophages via the scavenger receptor pathway. When LDL is incubated with DHA followed by gel filtration, LDL remains protected against subsequent Cu(2+)-induced oxidative modification, suggestive of stable modification of LDL in the presence of DHA. In contrast, DHA is ineffective against a metal ion-independent type of oxidative stress, viz. aqueous peroxyl radicals; under these conditions, only AA is able to inhibit lipid peroxidation in LDL. Our data indicate that vitamin C protects LDL against atherogenic modification by two different mechanisms that may act in concert: (i) free radical scavenging by AA prevents aqueous oxidants from attacking and oxidizing LDL, and (ii) stable modification of LDL by DHA or decomposition product(s) thereof imparts increased resistance to metal ion-dependent oxidation. PMID- 8419333 TI - Cell cycle regulation of a human cyclin-like gene encoding uracil-DNA glycosylase. AB - The predicted amino acid sequence of a human cDNA encoding uracil-DNA glycosylase activity shows striking similarity to the cyclin protein family. To characterize the expression of this DNA repair enzyme, we have isolated the corresponding genomic clone. This gene is contained within 4.2 kilobases and is composed of only two exons. Sequence analysis of the upstream region shows that it contains two cell cycle box (CCB) regulatory elements which are also found in yeast cyclin genes. Deletional analysis of the promoter reveals the presence of a repressor region located from position -812 to -603. An inverted CCB element (alpha-CCB) and an SP1-like binding site are contained within this region. When uracil-DNA glycosylase mRNA levels are examined in vivo, a 2-3-fold increase is associated with G1 phase in both HeLa S3 and WI38 cells. To examine the role of the 209-base pair repressor region in mediating cell cycle regulation, this fragment was used in gel shift assays with cellular extracts prepared from various stages of the cell cycle. Several specific complexes are formed during S and G2 phases which are not present during M and G1 phases. Two of the complexes are the result of alpha-CCB binding as they can be specifically disrupted by the addition of an oligonucleotide containing the alpha-CCB binding site. Immunoprecipitation studies reveal that uracil-DNA glycosylase protein levels are also elevated during G1 phase. Additionally, we show that the 36-kDa uracil-DNA glycosylase protein is turned over during the course of one cell cycle. These results demonstrate coordinate regulation of uracil-DNA glycosylase at both the transcriptional and the post-transcriptional levels as a function of the cell cycle. PMID- 8419334 TI - The tRNA-(m5U54)-methyltransferase of Escherichia coli is present in two forms in vivo, one of which is present as bound to tRNA and to a 3'-end fragment of 16 S rRNA. AB - The enzyme tRNA-(m5U54)-methyltransferase (EC 2.1.1.35) of Escherichia coli catalyzes the transfer of a methyl group from S-adenosyl-L-methionine to uridine in position 54 of the T psi-loop of all E. coli tRNA species, forming 5 methyluridine (m5U). In vivo, this enzyme is present both as a native polypeptide of 42 kDa and as a TrmA.RNA complex. The TrmA.RNA complex is not dissociated during strong denaturing conditions such as boiling in 8 M urea or 6 M guanidine HCl, consisting with that the RNA is covalently bound to the protein. After sequencing and Southern blot analyses, the RNA was identified to be a subset of undermodified tRNA species as well as the 3' terminus of 16 S rRNA. However, the complex is not associated with the ribosome and the covalently bound RNA does not affect the tRNA methylating activity of the enzyme. PMID- 8419335 TI - Structure of hepatitis B virus core and e-antigen. A single precore amino acid prevents nucleocapsid assembly. AB - The hepatitis B virus core gene codes for two polypeptides: the core protein, which assembles to form particles (HBcAg), and the secreted precore protein (HBeAg). Expression vectors directing the synthesis in Escherichia coli of a recombinant HBeAg corresponding in sequence to serum-derived HBeAg encompassing the 10 precore amino acids remaining after cleavage of the precursor and residues 1-149 of HBcAg (PC-HBeAg) were constructed. Recombinant PC-HBeAg, HBcAg, and C terminally truncated HBcAg were isolated from E. coli and analyzed by sucrose velocity sedimentation, electron microscopy, anti-HBc/e specific monoclonal antibody analysis, and for immunogenicity. HBcAg and truncated HBcAg formed 27-nm particles and displayed HBc antigenicity. In contrast, PC-HBeAg was nonparticulate and did not band in sucrose gradients. PC-HBeAg was recognized efficiently by HBeAg-specific antibodies and displayed little HBc antigenicity. Immunogenicity studies including T and B cell recognition confirmed that PC-HBeAg demonstrates HBe antigenicity. The presence of the 10 precore amino acids therefore prevented particle formation. To analyze which precore amino acids might be responsible for the prevention of particle formation a cysteine to glutamine substitution at amino acid position -7 was introduced into PC-HBeAg ( 7C-->Q)PC-HBeAg. This single amino acid change at position -7 restored particle formation and HBc antigenicity. The evolutionarily conserved cysteine at position -7 thus appears responsible for the prevention of particle assembly in the HBeAg biosynthesis pathway. PMID- 8419336 TI - Melanoma growth-stimulatory activity/GRO decreases collagen expression by human fibroblasts. Regulation by C-X-C but not C-C cytokines. AB - Melanoma growth-stimulatory activity (MGSA)/GRO is well characterized as a potent neutrophil chemoattractant. In the present study, we have demonstrated that MGSA induced a dose-dependent decrease in the expression of interstitial collagens by rheumatoid synovial fibroblasts. The decrease was observed over a dose range of 0.6-6.0 nM MGSA. This effect was specific, as MGSA had no demonstrable effect on the expression of collagen-degrading metalloproteinases, nor did it affect the collagenase inhibitor, tissue inhibitor of metalloproteinases. It also had no effect on the proliferation rate of these fibroblasts, unlike its mitogenic effect on melanoma cells. The ability to inhibit collagen expression was also demonstrated by another member of the C-X-C branch of the platelet factor 4 superfamily, interleukin-8 (IL-8), but not by RANTES, MIP-1 alpha, or MIP-1 beta, which belong to the C-C branch. Steady-state levels of expression of MGSA and IL 8 transcripts in normal adult tissues were dissimilar, suggesting that expression may be an important level at which the activity of these cytokines is regulated. Direct binding experiments with 125I-MGSA on synovial fibroblasts have allowed us to identify an MGSA receptor with a KD of 10.1 nM and approximately 75,000 binding sites/fibroblast. 125I-MGSA binding was specific and could not be displaced by unlabeled IL-8. These results suggest that MGSA, as well as IL-8, may play a role other than that of neutrophil chemo-attractant and more specifically, may be important in the regulation of collagen turnover. PMID- 8419337 TI - Involvement of Alu sequences in the cell-specific regulation of transcription of the gamma chain of Fc and T cell receptors. AB - The Fc epsilon RI-gamma chains are expressed in a variety of hematopoietic cells where they play a critical role in signal transduction. They are part of the high affinity IgE receptor in mast cells, basophils, Langerhans cells, and possibly other cells; a component of the low affinity receptor for IgG (Fc gamma RIIIA or CD16) in natural killer cells and macrophages; and part of the T cell antigen receptor in subsets of T cells. Here we have investigated the transcriptional regulation of the gamma chain gene by analyzing the 2.5-kilobase sequence upstream of the transcription start site. This sequence contains a promoter specific to cells of hematopoietic lineage. However, the tissue specificity of this promoter is only partial because it is active in all of the hematopoietic cells tested here, regardless of whether they constitutively express Fc epsilon RI- gamma chain transcripts. We have identified two adjacent cis-acting regulatory elements, both of which are part of an Alu repeat. The first (-445/ 366) is a positive element active in both basophils and T cells. The second ( 365/-264) binds to nuclear factors, which appear to be different in basophils and T cells, and acts as a negative element in basophils and as a positive one in T cells. Thus, this Alu repeat (90% identical to Alu consensus sequences) has evolved to become both a positive and negative regulator. PMID- 8419338 TI - Two mRNA transcripts (rBAT-1 and rBAT-2) are involved in system b0,(+)-related amino acid transport. AB - Previously, we isolated a cDNA clone (rBAT-1) of 2.2 kilobase pairs (kb) from a rabbit kidney cortex cDNA library, encoding a protein involved in sodium independent transport of L-dibasic amino acids, L-cystine, and some neutral amino acids via a system related to b0,(+)-like activity (Bertran, J., Werner, A., Moore, M. L., Stange, G., Markovich, D., Biber, J., Testar, X., Zorzano, A., Palacin, M., and Murer, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 5601-5605). In Northern blot hybridization using an rBAT-1 cDNA probe, 2.2- and 3.9-kb mRNA species were observed. Here we describe the isolation of a 3.9-kb cDNA clone (rBAT-2) by expression cloning using Xenopus laevis oocytes corresponding to the 3.9-kb mRNA species. On the basis of sequence analysis, in vitro translation (major protein of approximately 78 kDa), and functional analysis (expression of transport function), we conclude that rBAT-1- and rBAT-2-related proteins are identical: 677 amino acids in length, with most likely only one transmembrane spanning domain. There are seven differences in the nucleotide composition within a common overlap of 2189 nucleotides, resulting in 2 amino acid replacements. In comparison with rBAT-1, rBAT-2 has 26 additional nucleotides at the 5'-end, an identical location of the first polyadenylation signal, and approximately 1.7 kb of 3'-untranslated sequence (rich in AT(U) motifs) prior to a poly(A) tail of 63 adenines. We conclude that rBAT-1 and rBAT-2 encode the same protein and that the major difference seems to be related to the use of different polyadenylation signals. PMID- 8419339 TI - Mutational analysis of alpha-subunit of protein farnesyltransferase. Evidence for a catalytic role. AB - Protein farnesyltransferase from rat brain is composed of tightly associated alpha- and beta-subunits of 377 and 437 amino acids that migrate on SDS polyacrylamide gels with apparent molecular masses of 49 and 46 kDa, respectively. The enzyme attaches farnesyl groups to cysteines in p21ras and other proteins that contain cysteine residues at the fourth position from the COOH terminus. Production of stable enzyme in animal cells requires the simultaneous synthesis of both subunits, and all activity is lost when the subunits are dissociated chemically. The beta-subunit functions in the Zn(2+) dependent binding of the protein substrate. The role of the alpha-subunit is unknown. In the current studies we used in vitro mutagenesis and transfection of cloned cDNAs to define the parts of the alpha-subunit that are necessary to stabilize the beta-subunit and to support farnesyl transfer. Deletion of 51 amino acids at the NH2 terminus of the alpha-subunit allowed normal stabilization of the beta-subunit and production of normal enzyme activity, but deletion of 106 amino acids abolished both of these properties. A proline-rich region at residues 12-34 of the alpha-subunit is not required for activity, but its presence explains the anomalously slow migration of the polypeptide on SDS-polyacrylamide gels. Deletion of only 5 amino acids at the COOH terminus of the alpha-subunit reduced activity appreciably. Substitution of asparagine for a conserved lysine at position 164 produced an alpha-subunit that stabilized the beta-subunit normally and permitted normal binding of the two substrates, farnesyl pyrophosphate and p21H-ras. Nevertheless, the rate of transfer of the bound farnesyl group to p21H-ras was markedly reduced. The latter finding suggests that the alpha-subunit plays a direct role in the catalytic reaction in addition to its role in the stabilization of the beta-subunit. PMID- 8419340 TI - Cloning of human transketolase cDNAs and comparison of the nucleotide sequence of the coding region in Wernicke-Korsakoff and non-Wernicke-Korsakoff individuals. AB - Variants of the enzyme transketolase which possess reduced affinity for its cofactor thiamine pyrophosphate (high apparent Km) have been described in chronic alcoholic patients with Wernicke-Korsakoff syndrome. Since the syndrome has been shown to be directly related to thiamine deficiency, it has been hypothesized that such transketolase variants may represent a genetic predisposition to the development of this syndrome. To test this hypothesis, human transketolase cDNA clones were isolated, and their nucleotide and predicted amino acid sequence were determined. Transketolase was found to be a single copy gene which produces a single mRNA of approximately 2100 nucleotides. Additionally, the nucleotide sequence of the transketolase coding region in fibroblasts derived from two Wernicke-Korsakoff (WK) patients was compared to that of two nonalcoholic controls. Although nucleotide and predicted amino acid differences were detected between fibroblast cultures and the original cDNAs and among the cultures themselves, no specific nucleotide variations, which would encode a variant amino acid sequence, were associated exclusively with the coding region from WK patients. Thus, allelic variants of the transketolase gene cannot account for the biochemically distinct forms of the enzyme found in these patients nor be considered as a mechanism for genetic predisposition to the development of Wernicke-Korsakoff syndrome. Instead, the underlying mechanism must be extragenic and may be a result of differences in post-translational processing/modification of the transketolase polypeptide. PMID- 8419341 TI - The ductus arteriosus migratory smooth muscle cell phenotype processes tropoelastin to a 52-kDa product associated with impaired assembly of elastic laminae. AB - We established the identity of a 52-kDa protein secreted by fetal lamb ductus arteriosus (DA) smooth muscle cells (SMC) and suggest how it might be related to structural changes unique to DA development, i.e. reduced assembly of elastic laminae and associated formation of intimal cushions. We produced a monoclonal antibody (HI-20) to the 52-kDa protein and observed, by electron microscopy, immunogold labeling of elastin in both DA and aorta vessel walls. Western immunoblotting showed that HI-20, as well as antibodies to tropoelastin, reacted with the 52-kDa protein secreted by DA SMC, as well as with 68-kDa tropoelastin. The highly specific antibody to the carboxyl-terminal sequence of tropoelastin failed, however, to recognize the 52-kDa protein, although it reacted well with the 68-kDa tropoelastin. Amino acid analysis and sequencing data confirmed the identity of the affinity-purified 52-kDa protein as truncated tropoelastin with an intact amino terminus. Cell-free translation of mRNA extracted from DA and aorta SMC produced a 68-kDa, but not a 52-kDa, immunoprecipitated tropoelastin. When DA and aorta SMC were pulsed with [14C]valine, we immunoprecipitated, after only a 15-min chase, both 68-kDa and 52-kDa tropoelastin from cell extracts of DA SMC, but only the 68-kDa tropoelastin was present in aorta SMC. There was no evidence of proteolytic degradation of radiolabeled aorta 68-kDa tropoelastin to a 52-kDa species when mixed with DA SMC conditioned medium. This suggests that the 52-kDa tropoelastin is the result of cell-associated processing or degradation of an original 68-kDa product of translation. Furthermore, pulse chase experiments showed initial secretion of equivalent amounts of 68-kDa and 52 kDa tropoelastins by cultured DA SMC with increasing accumulation of the 52-kDa species, suggesting its impaired insolubilization. The production, in high concentration, of a 52-kDa tropoelastin product that lacks the carboxyl terminus, may prevent its alignment on the microfibrillar scaffold, resulting in abnormal assembly of elastic laminae in the DA. The accumulation of this soluble tropoelastin may be associated with the previously described property of chemotaxis resulting in the increased SMC migration into the subendothelium associated with DA intimal thickening. PMID- 8419342 TI - Mutational analysis of G protein alpha subunit G(o) alpha expressed in Escherichia coli. AB - G protein-mediated signal transduction is dependent on alpha subunit interactions with beta gamma subunits, receptors, effectors, magnesium ions, and guanine nucleotides. The interdependence of these interactions can be probed by mutational analysis. We developed large scale screening procedures in recombinant Escherichia coli to identify and characterize novel mutations in G(o) alpha. Random mutations were generated by polymerase chain reaction in the amino terminal 56 amino acids of G(o) alpha. Guanine nucleotide binding properties of the mutants were assayed in situ and in crude extracts of recombinant E. coli. beta gamma interactions were assayed by pertussis toxin mediated ADP ribosylation. Efficacy of the screening procedures was evaluated by studying properties of wild-type G(o) alpha and site-directed mutations that were characterized previously in other G proteins. Several novel mutants with altered GTP binding characteristics and reduced ability to interact with beta gamma had been isolated from the randomly generated mutant library. ADP-ribosylation of mutants R10G, K21N, and K35E was significantly reduced, whereas two of the mutants bearing multiple amino acid substitutions were refractory to modification. Mutant K35E also exhibited reduced affinity to guanosine 5'-(3-O thio)triphosphate at submicromolar concentrations of magnesium. These experiments demonstrate the feasibility of using large scale random mutagenesis in the studies of G protein function. PMID- 8419343 TI - Purification and properties of the RecR protein from Bacillus subtilis 168. AB - Genetic evidence suggests that the Bacillus subtilis recR gene product is involved in DNA repair and recombination. To assign a biochemical function to the recR gene product, the RecR protein was labeled and purified by monitoring the radioactive label. NH2-terminal protein sequence analysis of RecR was consistent with the deduced amino acid sequence of the recR gene. The RecR protein (molecular mass of 25 kDa, isoelectric point 5.4) bound single- and double stranded DNA in a filter binding assay. RecR-DNA complex formation is enhanced by the presence of a damage in the DNA substrate. The RecR-DNA complex formation proceeds in the absence of divalent cations and nucleotide cofactors, but is markedly stimulated by ATP and divalent cations. In our experimental conditions the apparent equilibrium constants of the optimized RecR-DNA complexes are 3 x 10(-7) M and 9 x 10(-7) M for damaged and undamaged DNA, respectively. The binding reaction is cooperative. Electron microscopy studies show that the presence of divalent cations increases the rate of RecR protein self-assembly. Addition of ATP or dATP promotes the organization of discrete series of quaternary structures on DNA, but ATP gamma S inhibits the DNA binding activity. A possible mechanism for the RecR function in DNA repair is discussed. PMID- 8419344 TI - In vitro fusion of rabbit liver Golgi membranes with liposomes. AB - Fusion of Golgi membranes isolated from rabbit liver with liposomes was studied by lipid mixing of fluorescent lipid analogues and internal content mixing and by electron microscopic observation of transfer of horseradish peroxidase from liposomes into Golgi membranes. A monoclonal antibody was used to confirm fusion of Golgi membranes but not other contaminating vesicles. Fusion was rapid and efficient, reaching about 20% of the maximum after a 5-min incubation using small or large unilamellar dioleoylphosphatidylcholine vesicles. The fusion was dependent on temperature, decreasing at lower temperatures, and becoming nearly zero below 10 degrees C. The addition of ATP, GTP, cytosolic factors, or N ethylmaleimide did not affect fusion. Treatments of Golgi membranes with 0.1 M Na2CO3 or 1 M KCl did not cause any changes in fusion. However, treatment with proteases inhibited fusion. These results suggest that Golgi integral membrane protein(s) are involved in fusion. Changing the medium to an isoosmotic substance, sucrose, in place of KCl or NaCl inhibited fusion. The binding assay of fluorescent liposomes to Golgi membranes showed that lowering the temperature or replacing salts with sucrose did not affect binding. However, treatment of Golgi membranes with proteases inhibited binding. Addition of phosphatidylserine or phosphatidylethanolamine to dioleoylphosphatidylcholine liposomes caused a 2 fold increase in binding and fusion. Fusion between Golgi membranes by themselves did not occur. These results provide some information on the mechanism of intracellular vesicular transport. PMID- 8419345 TI - Expression and structural studies of fasciclin I, an insect cell adhesion molecule. AB - Fasciclin I is a lipid-linked cell-surface glycoprotein that can act as a homophilic adhesion molecule in tissue culture cells. It is thought to be involved in growth cone guidance in the embryonic insect nervous system. To facilitate structure-function studies, we have generated Chinese hamster ovary (CHO) cell lines expressing high levels of cell surface grasshopper and Drosophila fasciclin I. Grasshopper fasciclin I released by phospholipase C cleavage was purified on an immunoaffinity column and single crystals were obtained that diffracted to approximately 5-A resolution. We also generated CHO and Drosophila S2 cell lines that produce a secreted form of fasciclin I. Fasciclin I expressed in S2 cells contains significantly less carbohydrate than the protein expressed in CHO cells, and may therefore be more suitable for crystallization. Biochemical characterization of purified fasciclin I indicates that the extracellular portion exists as a monomer in solution. Circular dichroism studies suggest that fasciclin I is primarily alpha-helical. Its structure is therefore different from other known cell adhesion molecules, which are predicted to be elongated beta-sheet structures. This suggests that fasciclin I may define a new structural motif used to mediate adhesive interactions between cell surfaces. PMID- 8419346 TI - Characterization of the forward and reverse integration reactions of the Moloney murine leukemia virus integrase protein purified from Escherichia coli. AB - The forward and reverse reactions for integration were characterized for the Moloney murine leukemia virus integrase (M-MuLV IN) protein. The M-MuLV IN was recombinantly produced in Escherichia coli, and was purified to greater than 90% homogeneity by a one-step affinity purification scheme. M-MuLV IN was highly active for integration as measured by in vitro cleavage and strand transfer assays. Furthermore, the integration of a model viral substrate into lambda concatamers by IN correctly produced the flanking 4-base pair duplications characteristic of M-MuLV IN. The reverse reaction of integration, disintegration, was also catalyzed by the recombinant M-MuLV IN. Two products were generated, a 3'-recessed long terminal repeat and a ligated target DNA, from a model integration-intermediate substrate in the presence of M-MuLV IN. The requirements and optimal conditions for maximal integration and disintegration activity for M MuLV IN were determined. The forward and reverse reactions required different concentrations of manganese ion and reductant. Salt was also titrated for the forward and reverse reactions. Sodium chloride inhibited integration, but had little affect on disintegration. Low concentrations of potassium chloride enhanced integration, but had no affect on disintegration. The dinucleotide cleavage, strand transfer, and the disintegration reactions each had a unique pH profile of activity. PMID- 8419347 TI - Hsp90 chaperonins possess ATPase activity and bind heat shock transcription factors and peptidyl prolyl isomerases. AB - Heat shock proteins of the 82-90 kDa class (hsp82 and hsp90) are abundant, conserved, and ubiquitous from prokaryotes to eukaryotes. Although proposed to be chaperones, they had not been reported to possess enzymatic activity until our recent observation that pure trypanosomatid hsp83 had potent ATPase activity (Nadeau, K., Sullivan, M., Engman, D., and Walsh, C. T. (1992) Protein Sci. 1, 970-979). We have now purified the hsp90 homolog from Escherichia coli (HtpG) and from Saccharomyces cerevisiae (hsp82) to homogeneity and observe ATPase activity with kcat values of 3 min-1 and 140 min-1. In addition, examinations of purified rat hsp90 and human hsp90 detect ATPase activity with a kcat of 0.6 min-1 and 10 min-1. Each of these hsp90s undergoes autophosphorylation on serine or threonine residues. In prokaryotes and eukaryotes, the induction of hsps during heat shock is controlled, respectively, by the binding of an alternate sigma 32 or a transcriptional activator (heat shock factor or HSF) at heat shock promoter elements. Here we show that E. coli HtpG immobilized to Affi-Gel beads selectively retains sigma 32 while the yeast hsp90 and rat hsp90 retain HSF. The peptidyl prolyl isomerase hsp59 of the FK506 binding class is known to bind to hsp90. We also detect binding of the other family of PPIases, the cyclophilins, to immobilized hsp90, consistent with a functional convergence of protein foldases. PMID- 8419348 TI - Lipopolysaccharide interaction with S2 subunit of pertussis toxin. AB - Using radioiodinated, photoactivable, reducible cross-linker conjugated bacterial endotoxic lipopolysaccharide (125I-ASD-LPS), we have demonstrated that LPS selectively binds to the S2 subunit of pertussis toxin (PT). Since LPS also interacts with the S2 subunit of the B-oligomer of the toxin, the binding of LPS to PT is not A-protomer (S1 subunit) dependent. The binding can be inhibited with native underivatized LPS and with purified lipid A, suggesting that the binding is mediated through the lipid A moiety of the LPS molecule. The binding of PT to LPS can be inhibited by bovine fetuin glycoprotein. Since PT has been demonstrated to interact specifically with N-linked oligosaccharide side chains of fetuin, the interaction of LPS with the S2 subunit of PT may involve carbohydrate-dependent interactions of the disaccharide backbone of lipid A with S2. Additional studies have documented that LPS binding to PT may be competitively inhibited by lysozyme but not by polymyxin B. Sequence analysis has allowed identification of a high degree of amino acid sequence similarity between the S2 subunit of PT and hen egg white lysozyme at the N-terminal 80-residue regions. Shared N-terminal sequence similarity between lysozyme, PT-S2, and a third LPS-binding protein alpha-lactalbumin allows tentative identification of a second family of LPS binding proteins. PMID- 8419349 TI - Biosynthesis of ubiquinone and plastoquinone in the endoplasmic reticulum-Golgi membranes of spinach leaves. AB - The localization of ubiquinone (UQ) and plastoquinone (PQ) biosynthesis in subfractions isolated from spinach leaves has been studied. UQ-9 and UQ-10 were found mainly in mitochondria, whereas PQ was enriched in chloroplasts, but also found in Golgi membranes. alpha-Unsaturated polyprenol-11 was also present at a low concentration in chloroplasts. Autoradiography revealed the presence of nonaprenyl-4-hydroxybenzoate (NPHB) and nonaprenyl-2-methylquinol (NPMQ) transferase activities involved in quinone biosynthesis in all subfractions, but the specific activities involved in quinone biosynthesis in the total microsomal fraction were 20 times higher than those in mitochondria and chloroplasts. The isolated Golgi vesicles were particularly enriched in both activities. When the incubation medium containing total microsomes or Golgi membranes was supplemented with NADH, NADPH, S-adenosylmethionine, and an ATP-generating system, NPHB and NPMQ were transferred to UQ-9 and PQ, respectively. trans-Prenyltransferase, which synthesizes the side chain of UQ and PQ, was present in the total microsomal fraction. With farnesyl-PP as substrate, no product was formed, but with geranyl-PP, solanesyl-PP was synthesized and transferred to 4 hydroxybenzoate present in the total microsomal fraction. The results show that these membranes from spinach contain farnesyl-PP synthetase. It is concluded that the plant leaf Golgi membranes contain the enzymes for both UQ and PQ biosynthesis and that a specific transport and targeting system is required for selective transfer of UQ to the mitochondria and of PQ to the chloroplast. PMID- 8419350 TI - Engineering mouse P450coh to a novel corticosterone 15 alpha-hydroxylase and modeling steroid-binding orientation in the substrate pocket. AB - The F209L mutation alters specificity of P450coh from coumarin 7-hydroxylation to 15 alpha-hydroxylation of 11-deoxysteroids such as testosterone and 11 deoxycorticosterone. Neither the wild-type nor F209L exhibits activity toward 11 beta-hydroxysteroids including corticosterone. Mutation of Phe-209 to Asn, however, confers on mutant F209N a high corticosterone 15 alpha-hydroxylase activity. F209V also exhibits low corticosterone 15 alpha-hydroxylase activity; Km and Vmax are 10-fold higher and lower, respectively, than for F209N. The results are consistent with the hypothesis that direct interaction of Asn-209 with 11OH is responsible for high corticosterone 15 alpha-hydroxylase activity. To support this hypothesis, a possible steroid-binding orientation is modeled in the substrate pocket of P450cam. Our weighted homology and constrained alignments map residue 209 of P450coh to Met-184 and Met-191 of P450cam. Energy minimization of corticosterone in the substrate pocket results in the 11OH of the steroid directed toward Met-184 (7 A) and Met-191 (16 A), and in C15 located near the sixth axial position of the heme. The steroid-binding model suggests that the P450cam's substrate pocket may be conserved in the mammalian P450 and can accommodate a steroid molecule, and that residue 209 appears to be located at the critical site that determines the steroid-substrate specificity of a P450 depending on the type of group at the 11-position of steroid molecule. PMID- 8419351 TI - Complex-dependent inhibition of factor VIIa by antithrombin III and heparin. AB - The regulation of the factor VIIa-tissue factor complex is essential for control of the hemostatic response. However, the role of the inhibitor antithrombin III in the regulation of factor VIIa has remained in question. The inhibition of factor VIIa activity by antithrombin III and heparin in the presence and absence of tissue factor was evaluated using the fluorescent substrate m-LGR-nds. Our data show that the activity of recombinant human factor VIIa is inhibited by antithrombin III in the presence of heparin at a rate of 1.7 x 10(2) M-1 s-1. In the presence of tissue factor, the rate constant for this reaction increases to 5.6 x 10(3) M-1 s-1. A 1:1 stoichiometric complex between factor VIIa and antithrombin III, with an apparent molecular weight of 110,000, was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A heterogeneous mixture of factor VIIa products with molecular weights between 50,000 and 80,000, most likely representing proteolytically degraded factor VIIa-antithrombin III complexes, was also observed. PMID- 8419352 TI - Activation of the Escherichia coli nitrate reductase (narGHJI) operon by NarL and Fnr requires integration host factor. AB - Integration host factor protein (IHF) was shown to be required for Fnr- and NarL dependent activation of the nitrate reductase (narGHJI) operon of Escherichia coli in response to nitrate availability and anaerobiosis. Using a narG-lacZ reporter fusion to evaluate narGHJI expression in vivo both the nitrate and anaerobic dependent controls were severely impaired in a himA mutant compared with the wild type strain. IHF was also required for Fnr-independent anaerobic control of narGHJI expression. In vitro, purified IHF protein was shown to bind to a narG promoter fragment with an apparent dissociation value of 5 nM by use of a gel shift assay. DNase I footprinting studies revealed that IHF protects a 37 base pair region centered 125 base pairs 5' of the narG transcription site. These studies suggest that the IHF protein performs a DNA bending function at the narG promoter to allow nitrate-dependent activation by the NarL regulatory protein, and second, it enhances the Fnr-dependent expression from the narG promoter under anaerobic cell growth conditions. A model whereby three transcriptional activators, NarL, IHF, and Fnr, induce expression of a sigma 70-dependent promoter for the narGHJI operon is discussed. PMID- 8419353 TI - GDP dissociation inhibitor prevents intrinsic and GTPase activating protein stimulated GTP hydrolysis by the Rac GTP-binding protein. AB - The majority of the GTP-binding proteins of the Ras superfamily hydrolyze GTP to GDP very slowly. A notable exception to this are the Rac proteins, which have intrinsic GTPase rates at least 50-fold those of Ras or Rho. A protein (or proteins) capable of inhibiting this GTPase activity exists in human neutrophil cytosol. Since Rac appears to exist normally in neutrophils as a cytosolic protein complexed to (Rho)GDI, we examined the ability of (Rho)GDI to inhibit GTP hydrolysis by Rac. (Rho)GDI produced a concentration-dependent inhibition of GTP hydrolysis by Rac1 that paralleled its ability to inhibit GDP dissociation from the Rac protein. Maximal inhibition occurred at or near equimolar concentrations of the GDI and the Rac substrate. The ability of two molecules exhibiting GTPase activating protein (GAP) activity toward Rac to stimulate GTP hydrolysis was also inhibited by the presence of (Rho)GDI. The inhibitory effect of the GDI could be overcome by increasing the GAP concentration to levels equal to that of the GDI. (Rho)GDI weakly, but consistently, inhibited GTP gamma S (guanosine 5'-3-O (thio)triphosphate) dissociation from Rac1, confirming an interaction of (Rho)GDI with the GTP-bound form of the protein. These data describe an additional activity of (Rho)GDI and suggest a mechanism by which Rac might be maintained in an active form in vivo in the presence of regulatory GAPs. PMID- 8419354 TI - Thrombin-variable region 1 (VR1). Evidence for the dominant contribution of VR1 of serine proteases to their interaction with plasminogen activator inhibitor 1. AB - The importance of a specific variable region in different serine proteases for the interaction with plasminogen activator inhibitor 1 (PAI-1) is studied. To that end, we have constructed a thrombin substitution variant, thrombin-VR1, in which the entire variable region 1 (VR1) of the protease domain (Phe-34 to Leu 40) has been replaced by the corresponding sequence (Phe-294 to Phe-305) of tissue-type plasminogen activator. The substitution resulted in a 2000-fold increase of the second-order rate constant of inhibition by PAI-1 (k2 = 2.2 x 10(6) M-1 s-1) as compared to alpha-thrombin (k2 = 1.1 x 10(3) M-1 s-1). Inhibition of thrombin-VR1 by PAI-1 is mediated by the formation of SDS-stable, enzyme-inhibitor complexes. The substitution did not affect the rate constant of inhibition by antithrombin III, whereas clotting efficiency and the rate of inhibition by heparin cofactor II were decreased 3-fold. These results demonstrate the importance and specificity of the protease domain VR1 region for the interaction of PAI-1 with its target proteases. PMID- 8419355 TI - Controlling lipase stereoselectivity via the surface pressure. AB - In the present study, the stereoselectivity of Rhizomucor miehei lipase, lipoprotein lipase, Candida antarctica B lipase, and human gastric lipase towards racemic dicaprin spread as a monolayer at the air-water interface was investigated. For this purpose we have developed a method with which the enantiomeric excess of the residual substrate can be measured in monomolecular films. The stereoselectivity, which is one of the main aspects of enzymic catalysis, was found to depend on the surface pressure of the substrate. With all four lipases tested, low surface pressures enhanced the stereoselectivity while decreasing the enzymes' catalytic activity. PMID- 8419356 TI - Equine lutropin and chorionic gonadotropin bear oligosaccharides terminating with SO4-4-GalNAc and Sia alpha 2,3Gal, respectively. AB - Equine chorionic gonadotropin (eCG) and lutropin (eLH) are heterodimeric glycoprotein hormones which are synthesized in the placenta and pituitary, respectively. The beta subunits of eCG and eLH, like their alpha subunits, arise from a single gene and have identical amino acid sequences. In contrast, the beta subunits of CG and LH in primates arise from different genes and differ in sequence. We have examined the structures of the Asn-linked oligosaccharides on eCG and eLH. eCG bears di- and tri-branched Asn-linked oligosaccharides terminating with Sia alpha 2,3 or 6Gal beta 1,4GlcNAc. In contrast, > 72% of the Asn-linked oligosaccharides on eLH have 1 or 2 branches terminating with the sequence SO4-4-GalNAc beta 1,4GlcNAc. The nonsulfated oligosaccharides on eLH are neutral (6% of the total) or have branches terminating with sialic acid-Gal (22% of the total). Since the alpha and beta subunits of eCG and eLH both contain the tripeptide motif, Pro-Xaa-Arg/Lys, recognized by the glycoprotein hormone specific GalNAc-transferase found in pituitary, expression of the GalNAc- and sulfotransferases must differ in the placenta and pituitary. eLH, but not eCG, is bound by the hepatic endothelial cell receptor specific for the sequence SO4-4 GalNAc beta 1,4GlcNAc. As a result, eLH is removed from the circulation 5.7-fold more rapidly than eCG and is selectively localized to the liver. Since the major structural difference between eCG and eLH is in the terminal glycosylation of their Asn-linked oligosaccharides and this has a major impact on circulatory half life, it is likely that the difference in circulatory half-life defines the functional difference between eCG and eLH. PMID- 8419357 TI - The roles of His-64, Tyr-103, Tyr-146, and Tyr-151 in the epoxidation of styrene and beta-methylstyrene by recombinant sperm whale myoglobin. AB - Previous studies demonstrated that at least two mechanisms are involved in the epoxidation of styrene and stilbene by myoglobin and H2O2 (Ortiz de Montellano, P. R., and Catalano, C. E. (1985) J. Biol. Chem. 260, 9265-9271). One mechanism, reaction of the olefin with the ferryl oxygen, preserves the olefin stereochemistry and incorporates an oxygen from H2O2 into the epoxide. The second mechanism, proposed to be a protein-mediated co-oxidation process, results in loss of stereochemistry and incorporation of an atom of molecular oxygen. To examine the role of individual residues in olefin epoxidation, we have examined the catalytic activities of the possible Tyr-->Phe mutants, the His-64-->Val mutant, and a protein combining all the tyrosine and histidine mutations. The latter protein is less stable than the other mutants and is the only one for which a protein radical is not detected in the reaction with H2O2. The Km and Vmax for styrene epoxidation range, respectively, from 0.3-8 mM and 12-35 pmol/min/nmol of protein. Incubation with H(2)18O2 results in 20-30% incorporation of labeled oxygen into the epoxide with all the mutants except Y103F/Y151F, Y146F/Y151F, H64V, and H64V/Y103F/Y146F/Y151F, for which 52, 58, 89, and 96% of the epoxide oxygen, respectively, is labeled. Oxidation of cis-beta methylstyrene by wild-type myoglobin yielded a 54:46 ratio of cis- and trans-beta methylstyrene oxides. The cis-isomer accounts for 47-100% of the epoxide produced by the mutant hemoproteins, with the two H64V mutants yielding almost exclusively the cis-epoxide. The oxygen in the cis-epoxide derives primarily or exclusively from H2O2 and that in the trans-epoxide from an alternative source. These results indicate that tyrosine residues may participate in, but are not essential for, protein-mediated epoxidation. In contrast, His-64 appears to be essential for co oxidative epoxidation because in its absence olefin epoxidation is mediated almost exclusively by ferryl oxygen transfer. PMID- 8419358 TI - Protein, cDNA, and genomic DNA sequences of the towel gourd trypsin inhibitor. A squash family inhibitor. AB - Two trypsin inhibitor components of the squash family were isolated and purified from the juice of the towel gourd (Luffa cylindrica) using anhydrotrypsin affinity chromatography followed by high pressure liquid chromatography. The inhibitors were sequenced and found to consist of 28 and 29 amino acid residues. The determined sequences show high similarity to other inhibitors of the squash family, especially in the location of disulfide bonds and the reactive site and also in the COOH-terminal region. A cDNA library of towel gourd was constructed and used as a template for polymerase chain reaction amplification of two cDNA fragments of the inhibitor with an overlapping sequence. A full-length cDNA sequence coding for the inhibitor was then completed. The open reading frame codes for a prepro-inhibitor protein with the pre- and pro-peptides consisting of 21 and 13 residues, respectively. The deduced amino acid sequence of 29 residues for the inhibitor is consistent with that determined by primary structure analysis. The genomic sequence of the mature inhibitor was also ascertained using the total DNA of the towel gourd as a polymerase chain reaction template. The genomic sequence is completely identical with that of the cDNA, showing no intervening sequence. PMID- 8419359 TI - Escherichia coli fumarate reductase frdC and frdD mutants. Identification of amino acid residues involved in catalytic activity with quinones. AB - Escherichia coli fumarate reductase (FRD) is a four-subunit enzyme that catalyzes the terminal step in anaerobic respiration to fumarate. The hydrophobic FrdC and FrdD subunits anchor the FrdA and FrdB catalytic subunits to the inner surface of the cytoplasmic membrane and are required for the enzyme to interact with quinones. Thirty-five single-site mutations were constructed in the FrdC and FrdD polypeptides by site-directed mutagenesis. Each mutant enzyme was characterized for its ability to catalyze quinone oxidation and reduction and to support growth of E. coli DW35 (delta frdABCD sdhC::kan) under selective conditions requiring functional enzyme. Replacement of FrdCE29 with Asp, Leu, Lys, or Phe had a deleterious effect both on quinol oxidase and quinone reductase activities. Substitution of FrdCH82 with Arg, Leu, Tyr, or Glu also decreased menaquinol oxidase activity, but had variable effects on the reverse reaction, the reduction of ubiquinone. Data are presented to support the hypothesis that the positive charge at FrdCH82 is required for stabilization of the quinone radical intermediate and the negative charge at FrdCE29 for deprotonation of menaquinol. Other critical amino acids identified in FrdC included Ala-32, Phe-38, Trp-86, Phe-87, and in FrdD residues Phe-57, Gln-59, Ser-60, and His-80. The established roles of such residues in the QA and QB sites of the photosynthetic reaction center would suggest a similar type of structure operative in the FRD complex. In such a model, Glu-29, Ala-32, His-82, Trp-86 of FrdC and His-80 of FrdD are considered participants in a QB-type site, and FrdD Phe-57, Gln-59, and Ser-60 components in an apolar QA-type site. PMID- 8419360 TI - Distribution of prenyltransferases in rat tissues. Evidence for a cytosolic all trans-geranylgeranyl diphosphate synthase. AB - The present study describes the presence of two different geranylgeranyl diphosphate (GGPP) synthase activities, one cytosolic and one membrane associated, in a number of rat tissues. Both enzymes utilize farnesyl diphosphate (FPP) and isopentenyl diphosphate (IPP) as substrates, but they give rise to different products. The membrane-associated activity produces trans,trans,cis (E,E,Z)-GGPP, involved in the biosynthesis of long-chain polyprenols. The cytosolic activity produces only the all-trans-(E,E,E) isomer of GGPP, which is utilized as substrate in cytosolic protein prenylation reactions. All-trans-GGPP synthase activity was recovered in the cytosolic fraction from all tissues investigated, but the specific activities varied. The highest specific activities were found in brain, spleen, and testis, followed by kidney and liver. The enzyme activity in rat brain cytosol was further characterized and found to exhibit a narrow pH optimum around 5.0-6.0 and to be highly stimulated by Zn2+. Maximal stimulation was attained with 1 mM Zn2+, whereas Mg2+ had no effect on the enzyme activity. The all-trans-GGPP synthase activity exhibited high affinities for its substrates, i.e. the apparent Km values for FPP and IPP were found to be 0.6 and 3.5 microM, respectively. When rats were fed mevinolin (lovastatin), FPP and all trans-GGPP synthase activities were affected differently in certain tissues. Mevinolin treatment resulted in an increase in FPP but a decrease in all-trans GGPP synthase activity in rat liver and kidney. In spleen mevinolin treatment caused a greater than 70% decrease in all-trans-GGPP synthase activity, while FPP synthase was almost unaffected. The presence of two different GGPP synthase activities in the cell, together with the fact that FPP and all-trans-GGPP synthesis in the cytosol are regulated independently, may be of significance in the regulation of isoprenoid biosynthesis, as well as of protein isoprenylation. PMID- 8419361 TI - Molecular nature of phospholipases A2 involved in prostaglandin I2 synthesis in human umbilical vein endothelial cells. Possible participation of cytosolic and extracellular type II phospholipases A2. AB - A lysate of unstimulated human umbilical vein endothelial cells (HUVEC) exhibited phospholipase A2 (PLA2) activity, which hydrolyzed phospholipids bearing arachidonate more preferentially than those bearing linoleate at the sn-2 position. An anti-rabbit cytosolic PLA2 monoclonal antibody absorbed the activity, whereas an anti-human type II PLA2 monoclonal antibody did not. HUVEC treated with thrombin generated prostaglandin I2 (PGI2), and the PLA2 activity of the thrombin-stimulated cells was absorbed almost completely by the anti cytosolic PLA2 antibody. HUVEC treated with tumor necrosis factor (TNF) also generated PGI2. PGI2 generation by TNF-treated cells was suppressed partially by extracellular addition of the anti-type II PLA2 antibody. PLA2 activity in a lysate of TNF-stimulated cells was increased about 2-3-fold, and about half of the increased activity was suppressed by the anti-type II PLA2 antibody. Addition of heparin together with TNF resulted in release of type II PLA2 in the medium. Thus, both cytosolic and type II PLA2s may be involved in agonist-stimulated PGI2 synthesis in HUVEC. Furthermore, exogenously added type II PLA2 was bound to the cell surface and synergistically enhanced PGI2 generation in TNF-stimulated HUVEC. This binding was blocked by either heparin or a monoclonal antibody recognizing the heparin-binding domain of type II PLA2. Taken together, type II PLA2 generated endogenously as well as added exogenously may be captured on the HUVEC surface via heparan sulfate proteoglycan and may contribute to cellular arachidonate metabolism. PMID- 8419362 TI - Mutant strains of Saccharomyces cerevisiae lacking sphingolipids synthesize novel inositol glycerophospholipids that mimic sphingolipid structures. AB - Mutant strains of Saccharomyces cerevisiae, termed SLC, make no detectable sphingolipids when cultured without a sphingolipid long chain base such as phytosphingosine (Dickson, R.C., Wells, G.B., Schmidt, A., and Lester, R.L. (1990) Mol. Cell. Biol. 10, 2176-2181). When grown with phytosphingosine they make sphingolipids in normal amounts. SLC strains carry a secondary suppressor mutation that bypasses the need to synthesize a long chain base. Hypothetically, the suppressor mutation could function by mutating a gene whose protein product required a sphingolipid for function, by increasing the level of one or more endogenous lipids, or by making new lipid(s). Here we demonstrate that SLC strains make novel inositol glycerophospholipids when cultured without a long chain base. The novel lipids are phosphatidylinositol (PI), mannosyl-PI, and inositol-P-(mannosyl-PI), containing 1 mol of C26 fatty acid, ordinarily found in yeast sphingolipids but not usually found in glycerophospholipids; the C26 fatty acid appears to be located at the SN-2 position of the glycerol. In addition, mono-fattyacyl versions of these three compounds were also identified. The polar head groups and hydrophobic portions of these novel lipids are strikingly similar to S. cerevisiae sphingolipids found in wild-type cells. We speculate that the novel lipids structurally mimic sphingolipids and thereby compensate for some sphingolipid function(s) necessary for growth. PMID- 8419363 TI - Glycosylation of human corticosteroid-binding globulin at aspargine 238 is necessary for steroid binding. AB - Human corticosteroid binding-globulin (CBG) is a plasma glycoprotein that binds and regulates the biological activity of glucocorticoids and progesterone. Carbohydrates comprise approximately 25% of its molecular mass being represented by bi- and triantennary N-linked oligosaccharides of the N-acetyllactosamine type. To assess the impact of these carbohydrate chains on CBG production and steroid binding, we mutated a human CBG cDNA so that the six consensus sites for N-glycosylation in the CBG polypeptide were eliminated individually and in various combinations. Expression of the mutant cDNAs in Chinese hamster ovary cells showed that all consensus sites may be utilized during the CBG biosynthesis and that the immunochemical properties of the recombinant glycoproteins are similar to those of CBG isolated from human serum. Removal of sugar chains generally led to a reduction in the secretion of recombinant CBG, but complete removal of N-glycosylation sites did not prevent production or secretion of the protein. Our data indicate that an oligosaccharide linked to Asn238 is essential for steroid binding, and we suggest that an interaction between this sugar chain and the polypeptide may be essential for the creation of a high affinity steroid binding site. In addition, concanavalin A chromatography of mutants containing only one N-glycosylation site at either Asn74 or Asn238 indicated that processing of the oligosaccharides at these positions is site-specific. PMID- 8419364 TI - The gamma subunit of the Escherichia coli ATP synthase. Mutations in the carboxyl terminal region restore energy coupling to the amino-terminal mutant gamma Met-23 ->Lys. AB - The gamma subunit mutations, gamma Met-23-->Lys or Arg, in the Escherichia coli ATP synthase were previously reported to cause dramatically inefficient energy coupling between ATPase catalysis and H+ translocation (Shin, K., Nakamoto, R.K., Maeda, M., and Futai, M. (1992) J. Biol. Chem. 267, 20835-20839). In this paper, we report that second-site mutations in the gamma subunit can suppress the effects of gamma Met-23-->Lys. By screening randomly mutagenized uncG (gamma Met 23-->Lys), eight mutations in the carboxyl-terminal region were identified; strains carrying gamma Arg-242-->Cys, gamma Gln-269-->Arg, gamma Ala-270-->Val, gamma Ile-272-->Thr, gamma Thr-273-->Ser, gamma Glu-278-->Gly, gamma Ile-279- >Thr, or gamma Val-280-->Ala in combination with gamma Met-23-->Lys were able to grow by oxidative phosphorylation. H+ pumping assayed in membranes prepared from double mutation strains demonstrated that efficient ATP-dependent H+ transport was restored. Interestingly, the single mutations, gamma Gln-269-->Arg or gamma Thr-273-->Ser, caused reduced growth by oxidative phosphorylation; however, when these mutations were in combination with gamma Met-23-->Lys, growth was substantially increased. Furthermore, strains carrying gamma Met-23-->Lys, gamma Gln-269-->Arg, or gamma Thr-273-->Ser as single mutations were temperature sensitive, whereas, strains with the double mutations, gamma Met-23-->Lys/gamma Gln-269-->Arg or gamma Met-23-->Lys/gamma Thr-273-->Ser, were thermally stable. Taken together, these results strongly suggest that gamma Met-23, gamma Arg-242, and the region between gamma Gln-269 to gamma Val-280 are close to each other and interact to mediate efficient energy coupling. PMID- 8419365 TI - Purification and characterization of SAR1p, a small GTP-binding protein required for transport vesicle formation from the endoplasmic reticulum. AB - SEC12 encodes an integral membrane glycoprotein essential for vesicle formation from the endoplasmic reticulum (ER) in yeast. The SAR1 gene was discovered as a multicopy suppressor of a sec12ts strain and encodes a 21-kDa GTP-binding protein also required for protein transport from the ER to the Golgi apparatus (Nakano, A., and Muramatsu, M. (1989) J. Cell Biol. 109, 2677-2691). We have purified Sar1p to apparent homogeneity from cells harboring a galactose-regulated recombinant SAR1. Purified Sar1p binds guanine nucleotides specifically and exhibits GTPase activity (0.001 min-1). Nucleotide exchange and hydrolysis rates are greatly increased in the presence of Mg2+ and nonionic detergents or phospholipids. An assay that measures the formation of a vesicle intermediate in ER to Golgi transport was devised that is dependent on the addition of purified Sar1p. This assay employs membranes prepared from wild-type cells and cytosol fractions depleted of Sar1p due to overproduction of Sec12p or by gel filtration chromatography. The gel-filtered cytosol requires the addition of Sar1p and GTP to support vesicle budding. Sar1p prebound with GTP gamma S inhibits Sar1p function in the vesicle formation assay. The results indicate a role for Sar1p in vesicle budding from the ER and suggest that GTP hydrolysis by Sar1p is required for this event. PMID- 8419367 TI - The 5'-terminal region of the apocytochrome b transcript in Crithidia fasciculata is successively edited by two guide RNAs in the 3' to 5' direction. AB - We analyzed the chimeric guide RNA (gRNA)-mRNA molecules in Crithidia fasciculata that are predicted to transiently exist in editing of the 5'-terminal domain of apocytochrome b (CYb) mRNA, by polymerase chain reaction amplification and DNA sequencing, and obtained evidence suggesting that among the 14 editing sites numbered from 3' to 5', the sequence in the 3'-half of the sites (3' block) was specified by one guide RNA species (gRNA-I) and that in the remaining half of the sites (5' block) by the other guide RNA species (gRNA-II) and that the direction of editing in each block was 3' to 5'. The predicted transition site of editing by two gRNAs was between the first and second U residues from the 3' end within editing site 7. We found that a stretch of the edited sequence in the 3' block of mRNA could form a stable duplex with a stretch immediately upstream of the guide sequence in gRNA-II. The result leads to a successive editing model that the 3' block of pre-edited mRNA is first edited by gRNA-I, and after completion of editing, the 5' portion of gRNA-II pairs with the edited mRNA for editing of the 5' block. PMID- 8419366 TI - Site-directed mutagenesis of the NH2 terminus of T4 endonuclease V. The position of the alpha NH2 moiety affects catalytic activity. AB - Reductive methylation of the alpha NH2 moiety of the DNA repair enzyme T4 endonuclease V has been shown previously to eradicate both the N-glycosylase and apyrimidinic/apurinic lyase activities of the enzyme (Schrock, R. D., III, and Lloyd, R. S. (1991) J. Biol. Chem. 266, 17631-17639). The present study uses the technique of site-directed mutagenesis to investigate the important parameters involved in the cleavage mechanism. The prediction was that the addition of an amino acid in the immediate NH2-terminal region of the protein would alter the proximity of the alpha NH2 moiety of Thr2 to its target, thereby severely compromising the enzyme's catalytic activity. However, substitutions in this region generally should be tolerated. To test this hypothesis, three substitutions of the NH2-terminal amino acid were produced: Ser2 (T2S), Val2 (T2V), and Pro2 (T2P). An addition mutant was also produced by adding a glycine between the first and second amino acids of the protein (Thr2-Gly-Arg3) (+Gly). The T2P and +Gly mutants had negligible pyrimidine dimer-specific N-glycosylase activity as well as negligible pyrimidine dimer-specific nicking activity in vitro. Conversely, the T2S enzyme exhibited wild type levels of activity and the T2V exhibited intermediate levels of activity in vitro. Results from ultraviolet (UV) survival studies of the mutant enzymes indicated that the in vivo activities of these enzymes were directly correlated to the enzymes' ability to cleave at pyrimidine dimers in vitro. These results indicate that a critical parameter for the functionality of endonuclease V is the relative distance between the primary alpha NH2 group in the active site of the enzyme and those elements responsible for DNA binding and pyrimidine dimer recognition. PMID- 8419368 TI - Amino acid substitutions at position 47 of human beta 1 beta 1 and beta 2 beta 2 alcohol dehydrogenases affect hydride transfer and coenzyme dissociation rate constants. AB - Human liver alcohol dehydrogenase isoenzymes beta 1 beta 1 and beta 2 beta 2, in which position 47 in the coenzyme binding domain is an arginine or histidine, respectively, differ remarkably in steady-state kinetics. To understand which catalytic steps affect these kinetics, apparent coenzyme dissociation and association rate constants, and apparent 4-trans-(N,N dimethylamino)cinnamaldehyde (DACA) hydride transfer rate constants were obtained with stopped-flow kinetics. Enzymes containing site-specific mutations of Arg-47 in beta 1 beta 1 (beta 47R) to His (beta 2 beta 2 or beta 47H), Lys (beta 47K), or Gln (beta 47Q) were studied. Apparent coenzyme dissociation rate constants are greatly affected by substitutions at position 47, in which mutant enzymes with a weak base or a neutral residue at this position (beta 47H and beta 47Q) exhibit faster rate constants than beta 47R and beta 47K. Substitutions at position 47 have less effect on apparent coenzyme association rate constants. The kinetics of NADH association for beta 47H and beta 47Q are consistent with a two-step mechanism in which the bimolecular binding step is coupled to a unimolecular process. These findings indicate that the greater role of position 47 in coenzyme dissociation may occur after a coenzyme-induced isomerization. Substitutions at position 47 also strongly influence apparent DACA hydride transfer rate constants; hydride transfer is faster with mutant enzymes containing weak bases like histidine at this position. Steady-state kinetics, however, reveal that the rate-limiting step of both beta 47R and beta 47H for acetaldehyde reduction and for ethanol oxidation is coenzyme product dissociation. Thus, the different activities of beta 1 beta 1 and beta 2 beta 2 for ethanol oxidation and acetaldehyde reduction are caused primarily by different coenzyme dissociation rates. PMID- 8419369 TI - Genetic engineering of snake toxins. Role of invariant residues in the structural and functional properties of a curaremimetic toxin, as probed by site-directed mutagenesis. AB - To study the site by which erabutoxin a (Ea) from Laticauda semifasciata binds to the nicotinic acetylcholine receptor, we mutated most residues that are shared with other curaremimetic toxins and studied the structural and biological consequences of introduced mutations. By site-directed mutagenesis, we changed Ser-8 into Gly (EaS8G), Lys-27 into Glu (EaK27E), Trp-29 into Phe (EaW29F) and His (EaW29H), Asp-31 into His (EaD31H), Phe-32 into Leu (EaF32L), Arg-33 into Lys (EaR33K) and Glu (EaR33E), Gly-34 into Ser (EaG34S), Glu-38 into Gln (EaE38Q) and Lys (EaE38K), Gly-49 into Val (EaG49V), and Leu-52 into Ala (EaL52A). All mutants were homogeneous as judged by various analytical procedures. EaE38Q, EaG49V, and EaL52A bound the nicotinic acetylcholine receptor with apparent Kd values close to 10(-10) M, virtually identical to wild Ea. Therefore, Glu-38, Gly-49, and Leu 52 are not important elements in the expression of curaremimetic function in Ea. Mutations of Phe-32 and Gly-34 provoked a 7-fold affinity decrease, suggesting that these residues moderately contribute to function. The 176-fold affinity decrease due to mutation of Ser-8 may reflect some structural change that operates in the polypeptide chain of the mutant, as detected by circular dichroism. Decreases in affinity by a factor of 175, 67, 46, and 318 were seen upon mutations of Lys-27 into Glu, Trp-29 into Phe, Asp-31 into His, and Arg-33 into Glu, with no concomitant change in secondary structure. These residues appear to be important elements of the curaremimetic function of Ea. Thus, a picture of the contribution of conserved residues to the function of a curaremimetic toxin is proposed on the basis of experimental evidence. PMID- 8419370 TI - Structural and functional domains of Escherichia coli ribosomal protein L7/L12. The hinge region is required for activity. AB - Variant forms of Escherichia coli ribosomal protein L7/L12 were constructed, overexpressed, and purified. These included proteins that deleted residues 35-52 (delta 35-52) and 42 to 52 (delta 42-52), others that contained single cysteine substitutions at residues 63 and 89, and combinations of the deletions and cysteine substitutions. Chemical modification of the introduced cysteine residues with [14C]iodoacetamide was used to radiolabel the protein variants in order to quantify their binding to the ribosome. Neither of the deletions in the hinge domain, delta 35-52 and delta 42-52, had any effect on L7/L12 dimer formation as detected by cross-linking by dimethyl suberimidate. Perpendicular urea gradient gel electrophoresis showed that both deletion variants retained a compact structural element attributable to the globular C-terminal domain. Reconstitution of core particles depleted of wild type L7/L12 with the deletion proteins showed that delta 42-52 bound normally in 4 copies per particle, whereas delta 35-52 bound in only 2.5 copies following isolation of the particles by high speed centrifugation or gel filtration. Ribosomes mixed with an excess of the deletion variants and assayed directly for polyphenylalanine synthesis were completely inactive. The results suggest that the flexibility conferred by the hinge is required for activity, perhaps by allowing the C-terminal domain to occupy a location near the base of the L7/L12 stalk. PMID- 8419371 TI - Kinetic and structural analysis of the Mg(2+)-binding site of the guanine nucleotide-binding protein p21H-ras. AB - The coordination and binding of the Mg2+ ion in the nucleotide-binding site of p21 have been investigated using site-directed mutagenesis, kinetic methods, and phosphorous NMR. Mg2+ in the p21.nucleotide.Mg2+ complex appears to be in fast equilibrium with the solvent. The dissociation constant between Mg2+ and the p21.GDP complex was determined to be 2.8 microM. It decreases 30- or 16-fold on substituting Ser-17 or Asp-57 with alanine, respectively, whereas the T35A mutation has no effect. All three mutations influence the dissociation constants and the association and dissociation rate constants of the interaction between guanine nucleotides and p21, but to a different degree. We conclude that Thr-35 is only complexed to Mg2+ in the GTP conformation and both Asp-57 and Ser-17 appear to be critical for both GDP and GTP binding. 31P NMR spectra of the GDP and Gpp(NH)p (guanosine-5'-(beta,gamma-imido)triphosphate) complexes of mutated p21 show a remarkable perturbation of the guanine nucleotide-binding site compared to wild-type protein. The mutant proteins show reduced GTPase rates, which are not stimulated by the GTPase-activating protein GAP. p21(S17A) has been reported to function just as p21(S17N) as a dominant negative inhibitor of normal p21. We find that it inhibits oncogenic p21-induced survival of primary neurons. PMID- 8419372 TI - Activation of endothelial cell phospholipase D by hydrogen peroxide and fatty acid hydroperoxide. AB - We have investigated oxidant-mediated stimulation of phospholipase D (PLD) activity in bovine pulmonary artery endothelial cells (BPAEC), prelabeled with [32P]orthophosphate or [32P]lysophospholipids. Treatment of cells incubated in Hanks' balanced salt solution (HBSS) containing 0.5% ethanol with hydrogen peroxide (H2O2) or linoleic acid hydroperoxide (18:2-OOH) enhanced the formation of 32P-labeled phosphatidylethanol (PEt) and phosphatidic acid (PA) in a dose- and time-dependent manner, indicating the activation of PLD. The H2O2- and 18:2 OOH-mediated PLD activation was not associated with cytotoxicity as determined by [3H]deoxyglucose release. The addition of ferrous chloride (50 microM) augmented H2O2-induced formation of [32P]PEt and [32P]PA about 2-fold, whereas the addition of the iron chelator desferoxamine blocked the potentiating effect of ferrous chloride. Replacement of the HBSS medium with Medium 199 containing 20% calf serum also potentiated the effect of H2O2-induced PLD activation. In addition to phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI) were readily hydrolyzed by PLD in response to H2O2 and 18:2-OOH treatment. The substrate specificity for oxidant-stimulated PLD activity differed from that observed in the presence of bradykinin or exhibited by agonist stimulation with 12-O-tetradecanoylphorbol 13-acetate (TPA) where PC was the major phospholipid hydrolyzed by PLD. The formation of PEt in the presence of H2O2 and 18:2-OOH was not abolished by chelation of either extracellular Ca2+ with EGTA (5 mM) or intracellular Ca2+ with 1,2-bis-(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid acetoxymethyl ester (BAPTA-AM) (25 microM, 30 min). Furthermore, pretreatment of BPAEC with the protein kinase C (PKC) inhibitor staurosporine and down-regulation of PKC by chronic TPA treatment (100 nM, 18 hr) had no effect on H2O2-induced PLD activation, suggesting that PLD activation by H2O2 is independent of PKC activity. It is possible that H2O2- and 18:2-OOH-induced activation of PLD represents an important mechanism to produce PA and diacylglycerol in endothelial cells. PMID- 8419373 TI - A role for the mitochondrial Na(+)-Ca2+ exchanger in the regulation of oxidative phosphorylation in isolated heart mitochondria. AB - The role of the Na(+)-Ca2+ exchanger of heart mitochondria in cellular functioning is not yet clear. The objectives of this study were to investigate the effects of stimulation and inhibition of the Na(+)-Ca2+ exchanger on the matrix free Ca2+ concentration in isolated heart mitochondria and to determine the consequences of these changes on the rate of NADH production via Krebs cycle turnover and the oxidative phosphorylation rate (OPR) supported by alpha ketoglutarate dehydrogenase, a Ca(2+)-regulated matrix enzyme. Activation of Na(+)-Ca2+ exchange by increasing extramitochondrial Na+ concentration was found to decrease the matrix free [Ca2+] in a concentration-dependent manner. Inhibitors of mitochondrial Na(+)-Ca2+ exchange activity inhibited the decrease in matrix free [Ca2+] mediated by Na+. Increasing concentrations of Na+ were also found to inhibit both the rate of NADH production and OPR. Inhibitors of mitochondrial Na(+)-Ca2+ exchange also blocked the effects of Na+ on the rate of NADH production and OPR in a similar concentration range. The results indicate that alterations in matrix free [Ca2+] induced by changes in mitochondrial Na(+) Ca2+ exchange activity are translated into changes in the rate of NADH production and the overall rate of oxidative phosphorylation. PMID- 8419374 TI - Determinants of catalytic activity and stability of carbonic anhydrase II as revealed by random mutagenesis. AB - The functional importance of a conserved hydrophobic face in human carbonic anhydrase II (CAII), including amino acid residues 190-210, was investigated by random mutagenesis. The catalytic activity, inhibitor binding, and level of CAII expression in Escherichia coli of 57 single amino acid variants were measured revealing that the function of amino acids correlates with their secondary structure placement. Side chains of amino acids in beta-sheet structure are required for the formation of folded, stable protein while those in the turn region determine catalytic efficiency and inhibitor specificity. The CAII active site is extremely plastic, accommodating amino acid substitutions of varied size, charge, and hydrophobicity with little effect on catalysis; only substitutions at Leu198 and Thr199 decrease the rates of CO2 hydration and ester hydrolysis more than 5-fold. These results pinpoint the hydrogen bond network, including the zinc solvent molecule and Thr199, as crucial for high catalytic efficiency and also suggest that Leu198 forms a portion of a CO2 association site. Increased activity is observed for substitutions at Thr200 (esterase) and Leu203 (hydrase). In addition, the pKa of the zinc-bound water molecule varies upon substitution of amino acids which alter the overall charge of the active site. Three residues interact with sulfonamide inhibitors; substitutions at Thr199 decrease binding (up to 10(3)-fold) while mutations at Thr200 and Cys206 increase binding of dansylamide (up to 80-fold). Mutations in the beta-sheet structure (Asp190-Ser197 and Val207-Ile-210) decrease the protein expression of CAII in E. coli, causing the formation of insoluble protein aggregates in many cases. This may suggest an important role for these residues in the folding process. In addition, mutations in Trp192, cis-Pro202, and Trp209 increase thermal lability (up to 5000-fold). PMID- 8419375 TI - A major ubiquitin conjugation system in wheat germ extracts involves a 15-kDa ubiquitin-conjugating enzyme (E2) homologous to the yeast UBC4/UBC5 gene products. AB - In eukaryotes, conjugation of ubiquitin to proteins serves as a committed step for intracellular protein degradation. Formation of ubiquitin-protein conjugates involves the transfer of ubiquitin-conjugating enzyme (E2)-bound ubiquitin to the target proteins with or without the assistance of ubiquitin-protein ligase (E3). We report the isolation and characterization of an E2 purified from wheat germ that accounts for the majority of ubiquitin conjugation activity observed in vitro. This E2 is basic, has an apparent molecular mass of 15 kDa, and forms oligomers that dissociate upon treatment with sulfhydryl reducing agents. E(2)15kDa will not work alone in vitro but requires an additional factor putatively identified as an E3 for substrate recognition. This E3 is distinct from E3 alpha previously described to be required for N-terminal recognition of target proteins. Partial amino acid sequence analysis of E(2)15kDa revealed a substantial identity (approximately 80% in two peptide regions) with yeast E2s encoded by UBC4/UBC5 genes. This homology was confirmed by immunodetection of a 16-kDa yeast protein corresponding to the molecular mass of the UBC4/UBC5 proteins with E(2)15kDa antisera. The products of yeast UBC4 and UBC5 genes along with that of UBC1 gene constitute a subfamily of functionally overlapping E2s that mediate the selective degradation of short-lived and abnormal proteins in vivo. Considering the high degree of functional and structural similarity of wheat E(2)15kDa with that of yeast UBC4/UBC5, it is likely that yeast UBC4/UBC5 and their homologs from other eukaryotes exhibit the same E3 dependence in performing their roles in protein degradation. PMID- 8419376 TI - VMA12 is essential for assembly of the vacuolar H(+)-ATPase subunits onto the vacuolar membrane in Saccharomyces cerevisiae. AB - vma12 mutants of the yeast Saccharomyces cerevisiae, which were originally identified as calcium-sensitive (cls) mutants that were also respiratory deficient (Pet-), have a defect in vacuolar membrane H(+)-ATPase activity (Ohya, Y., Umemoto, N., Tanida, I., Ohta, A., Iida, H., and Anraku, Y. (1991) J. Biol. Chem. 266, 13971-13977). The VMA12 gene was cloned by complementation of the growth defects of vma12 mutants. The nucleotide sequence of the gene predicts a polypeptide of 215 amino acids (25.2 kDa) with two putative membrane-spanning domains. A null vma12 mutant, constructed by chromosomal deletion of the gene, is viable but has completely lost the vacuolar membrane H(+)-ATPase activity and exhibits the same growth defects as observed for the original vma12 mutants. Synthesis and targeting of the subunits of the H(+)-ATPase in the delta vma12 mutant cells were examined by Western blotting analyses of whole cell and vacuolar membrane protein extracts. None of the peripheral membrane subunits that we analyzed (the 69-, 60-, 42-, and 27-kDa subunits) was detected in the vacuolar membrane fractions, although the cellular levels of these polypeptides appeared to be normal. The 100- and 17-kDa integral membrane subunits of the enzyme were absent or present at a substantially reduced level in mutant vacuolar membrane fractions. Anti-Vma12p antibodies recognized a vacuolar protein with the expected molecular mass of 25 kDa. However, the Vma12 protein was not detected in the vacuolar membrane ATPase complex that had been solubilized with a zwitterionic detergent, ZW3-14, and purified by glycerol gradient centrifugation (Kane, P. M., Yamashiro, C. T., and Stevens, T. H. (1989) J. Biol. Chem. 264, 19236-19244). These results indicate that the VMA12 gene product is not a component of the active vacuolar ATPase complex and instead suggest that this protein is required during the process of assembly and/or targeting of the enzyme complex to the vacuolar membrane. PMID- 8419377 TI - NMR studies of the effect of hyperglycemia on intracellular cations in rat kidney. AB - Alterations in intracellular cation concentrations in the kidney during hyperglycemia may play a role in a number of complications associated with diabetes mellitus such as renal hypertrophy and hypertension. In this study, we have investigated the effect of 25 mM glucose on intracellular pH and free Mg2+, free Ca2+, and Na+ concentrations in the perfused kidneys of Sprague-Dawley rats using 31P, 19F, and 23Na triple-quantum filtered NMR. No significant alteration occurred in the intracellular free Mg2+ concentration, pH, or ATP concentration during hyperglycemia. However, a sizable (approximately 50%) increase in the intracellular Na+ concentration was inferred from 23Na triple-quantum filtered NMR after 30-45 min of perfusion with 25 mM glucose. Intracellular free Ca2+, measured to be 390 +/- 15 nM at 5 mM glucose, increased significantly (95%; p < 0.001) to a value of 765 +/- 28 nM after 30 min of perfusion with 25 mM glucose. This effect of glucose was reversible. Only small increases (< or = 20%) in free Ca2+ were found with addition of comparable concentrations of a nontransportable sugar (20 mM mannitol), indicating that glucose entry into the cell (through the Na+/glucose cotransporter) plays a role in causing the free Ca2+ increase. No effect on free Ca2+ was however, seen with 1 mM ouabain, which caused a sizable increase in intracellular Na+, indicating that Na+/Ca2+ exchange does not play a significant role in the maintenance of low intracellular free Ca2+ in the kidney and that the observed increase in free Ca2+ is probably due to a decrease in the Ca2+/Mg(2+)-ATPase activity during hyperglycemia. Increased concentrations of Na+ and free Ca2+ during hyperglycemia may be involved in the mechanism of renal hypertrophy and hypertension, frequently associated with diabetes mellitus. PMID- 8419378 TI - Regional dissemination of wear debris from a total knee prosthesis. A case report. PMID- 8419379 TI - Mesenchymoma of bone. A case report. PMID- 8419380 TI - Dermatitis complicating operatively induced anesthetic regions around the knee. A report of four cases. PMID- 8419381 TI - Cervical spondylotic myelopathy. PMID- 8419382 TI - Deformities of the forearm in patients who have multiple cartilaginous exostosis. AB - Thirty-five patients who had multiple cartilaginous exostosis and deformities of the wrist and forearm were evaluated to determine the natural history and prognostic factors for the deformities. Radial and ulnar shortening were correlated linearly (r2 = 0.86), showing that both bones of the forearm were symmetrically involved. The angular growth abnormality of the distal aspect of the radius was correlated with radial shortening (r2 = 0.53) and ulnar shortening (r2 = 0.61), but it had no correlation with negative variance of the ulnar (r2 = 0.30). Carpal slip was an independent factor, with no correlation with any of the factors studied. The ulnar tether theory for deformities of the wrist in patients who have multiple cartilaginous exostosis was not substantiated by this study. PMID- 8419383 TI - Infantile tibia vara. PMID- 8419384 TI - Early soft-tissue complications after fractures of the distal part of the radius. PMID- 8419385 TI - Survivorship analysis in total condylar knee arthroplasty. PMID- 8419386 TI - Failure of the porous-coated anatomic prosthesis in total knee arthroplasty due to severe polyethylene wear. AB - Four hundred and eighty-seven total knee arthroplasties were performed by a single surgeon with use of a porous-coated anatomic prosthesis between 1982 and 1989. There were thirty-two clinical failures (7 per cent) due to severe wear of the surfaces of the tibial and patellar polyethylene components. Thirty patients had a revision. The average time to failure of the implant was four and one-half years. The initial clinical symptoms of failure by wear consisted of a painless effusion with a decreased range of motion. Subsequent pain was considered as the criterion for failure necessitating operative intervention. Increased weight and decreased age of the patient and a thinner tibial component were significant predictors of an increased risk of failure (p < 0.01). Examination of retrieved tibial components revealed extensive delamination caused by fracture of the polyethylene at a depth of about one millimeter below the surface. Cracks that had propagated in from the medial and lateral peripheries of the tibial component toward the center of the condyles were also a common finding. It appears that the design of the implant as well as clinical factors (the age and weight of the patient) contributed to the mechanical failure of the polyethylene of these implants. PMID- 8419387 TI - Operative treatment of distal femoral fractures proximal to total knee replacements. AB - Twenty fractures of the distal part of the femur proximal to a total knee replacement were treated operatively by members of the New England Trauma Study Group. Notching of the anterior aspect of the femoral cortex was associated with only two of these fractures, and none of the knee prostheses was loose at the time of the fracture. All twenty fractures were treated with open reduction and stable internal fixation, and the operation on fifteen fractures was supplemented with bone grafts. Every fracture healed, and eighteen healed after a mean of sixteen weeks (range, six to forty weeks). Union of the other two fractures was delayed, but repeat open reduction and internal fixation combined with autogenous bone-grafting resulted in union. After operative treatment, the patients returned to the level of activity that they had had before the fracture. The pre-existing tibiofemoral alignment and range of motion of the knee were also restored. At the time of follow-up, the average clinical rating of the Knee Society for all twenty knees had not decreased compared with the score before the fracture. PMID- 8419388 TI - Arthroplasty of the metacarpophalangeal joints with use of silicone-rubber implants in patients who have rheumatoid arthritis. Long-term results. AB - The long-term results of 144 arthroplasties of the metacarpophalangeal joints, with use of silicone-rubber implants, were reviewed for twenty-seven patients (thirty-six hands) who had rheumatoid arthritis. The operations were all performed or supervised by the same surgeon. All patients had an arthroplasty of the metacarpophalangeal joint of all four fingers, and all had subjective and objective clinical evaluation and roentgenographic assessment. Relief of pain, the cosmetic appearance of the hand, the range of motion of the metacarpophalangeal and proximal interphalangeal joints, and the presence of residual deformity were evaluated. The average duration of follow-up was eight years and six months (range, five years to sixteen years and three months). Preoperatively, the mean active-extension deficit of the metacarpophalangeal joints ranged from 53 degrees (index finger, right hand) to 70 degrees (little finger, left hand) and the mean flexion ranged from 84 degrees (index finger, right hand) to 94 degrees (little finger, left hand). At the early follow-up evaluation (average, four months postoperatively; range, two to six months), the mean extension deficit had improved to a range of 7 degrees (little finger, right hand) to 19 degrees (index finger, left hand), and mean flexion ranged from 56 degrees (little finger, left hand) to 66 degrees (ring fingers). The range of motion had improved from a non-functional arc of flexion to a more functional arc of extension. At the time of the latest follow-up visit, the motion of the metacarpophalangeal joints had not deteriorated.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419389 TI - Effects of partial patellectomy and reattachment of the patellar tendon on patellofemoral contact areas and pressures. AB - We used a previously reported experimental method to measure patellofemoral contact areas and pressures in four pairs of human cadaveric knees before and after a partial patellectomy. The knee joints were loaded by application of a flexion moment, which was resisted by the extension moment of the quadriceps mechanism. Patellofemoral contact was measured with the use of pressure-sensitive film, at 30, 60, and 90 degrees of flexion of the knee. Partial patellectomy decreased the patellofemoral contact area and increased pressure. We observed alterations in the patterns of contact, including a proximal shift in patellofemoral contact, after partial patellectomy. An anterior reattachment of the patellar tendon significantly minimized the effects of 20 and 40 per cent patellectomies (p < 0.05). After a 60 per cent patellectomy, patellofemoral contact was altered markedly, with the contact area reduced to less than 50 per cent of the control values regardless of the position of the patellar tendon reattachment. PMID- 8419390 TI - Changes in residual volume relative to vital capacity and total lung capacity after arthrodesis of the spine in patients who have adolescent idiopathic scoliosis. AB - We evaluated pulmonary functions before correction and again after a mean follow up of three years in thirty-five patients who had adolescent idiopathic scoliosis. The mean age (and standard deviation) at the time of correction was 13.7 +/- 1.8 years, and at the time of follow-up it was 17.1 +/- 2.5 years. The findings in the patients were compared with those in matched normal control subjects. With the exception of forced vital capacity, all of the determinations of absolute pulmonary volume increased postoperatively, but the increases were not all proportional. When the preoperative and follow-up determinations were expressed as percentages of the predicted pulmonary volumes (on the basis of age) to eliminate any effects of the difference in age, there was no change in total lung capacity, but vital capacity and forced vital capacity were significantly reduced. In addition, there was a significant increase in residual volume. Of the mean increase in total lung capacity after correction of the scoliosis, 82 per cent was due to an increase in residual volume and 18 per cent, to an increase in vital capacity. However, in control subjects age-matched at the time of follow up, the increase in vital capacity contributed 69 per cent of the mean increase in total lung capacity, a very marked difference from the findings in the patients who had scoliosis. In addition, two pulmonary-volume ratios--residual volume to vital capacity and residual volume to total lung capacity--increased in a highly significant fashion (Mann-Whitney test, p < 0.001) after arthrodesis of the spine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419391 TI - Bone changes associated with intraosseous hypertension in the caprine tibia. AB - We investigated the effects of increased intraosseous pressure on new-bone formation in the proximal metaphysis of the caprine tibia. Intraosseous hypertension was produced by obstruction of venous outflow by ligation of the popliteal vein draining the proximal aspect of the tibia and occlusion of the medullary space with bone cement (groups 1 and 2). After the obstruction of venous outflow (day 0), the intraosseous pressure measured at the proximal tibial metaphysis increased significantly from a mean of 15.5 millimeters of mercury before the obstruction to a mean of 28.7 millimeters of mercury in groups 1 and 2. In group 1, obstruction of venous outflow was combined with intraosseous infusion of autogenous whole blood under pressure to maintain the intraosseous pressure between thirty and forty-five millimeters of mercury during days 0 through 5. The time for venous drainage was still prolonged and intraosseous pressures were still increased on days 5 and 10 (means, 26.8 and 26.2 millimeters of mercury, respectively) in groups 1 and 2. The intraosseous hypertension produced in group 1 was associated with a significant increase in periosteal (138 per cent), endocortical (369 per cent), and cancellous new-bone formation (889 per cent) at the tibial metaphysis compared with control values. Osseous necrosis within the metaphysis was not observed. PMID- 8419392 TI - Antibiotic prophylaxis with two doses of cephalosporin in patients managed with internal fixation for a fracture of the hip. AB - A prospective, randomized, double-blind study was performed to evaluate the effects of antibiotic prophylaxis on the development of a wound infection in 239 patients who had immediate stabilization of a fracture of the proximal part of the femur with a dynamic hip screw. The effects of two perioperative doses of cefotiam, given twelve hours apart, were compared with those of two doses of a placebo. Sixteen perioperative risk factors were evaluated to determine whether it was possible to identify patients who were at risk for a wound infection. All patients were followed for a minimum of six weeks. Antibiotic prophylaxis significantly reduced the prevalence of wound infection (p < 0.05): the rate of major wound infection decreased from 5 to 1 per cent and the rate of minor wound infection, from 11 to 4 per cent. The most powerful predictors of major wound infection were the duration of the operation, the interval between the accident and admission to the hospital, and the duration of postoperative urinary catheterization. The preoperative level of serum albumin and the absolute lymphocyte count were significant predictors (p < 0.05) of minor wound infection and systemic infection, respectively. PMID- 8419393 TI - The value of aspiration of the hip joint before revision total hip arthroplasty. AB - The role of aspiration of the hip joint before revision of a total hip arthroplasty remains controversial. To address this issue, we reviewed the results of 270 consecutive hips in which aspiration had been attempted before revision procedures that were performed between 1980 and 1988. All hips had intraoperative findings and clinical follow-up of at least two years to confirm the presence or absence of infection. Only six (2 per cent) of the 270 hips were determined to be infected. Aspiration had been attempted in all six hips, but fluid could be obtained from only four. All six hips also had clinical or radiographic signs, or both, of infection, including increasing pain within three years after the arthroplasty (four hips), inability of the patient to attain pain free status after the original procedure (four hips), radiographic findings compatible with infection (six hips), and a positive finding on culture of a specimen obtained from a previous aspiration (two hips). Because of these factors, aspiration was attempted a second time in four of the six hips and a third time in three of the four. The four hips from which fluid could be obtained had a total of ten successful aspirations; the cultures of specimens obtained from six of these procedures were positive and those from four were negative. The appearance of the capsular tissue at the time of the operation suggested infection in five of the six infected hips. Histological sections were positive for inflammation in all six: there was acute inflammation only in one, chronic inflammation only in two, and acute and chronic inflammation in three. No organisms were seen on gram stains of specimens from any of the six infected hips. Of the 254 hips that did not have an infection and had been aspirated successfully, thirty-two (13 per cent) had a false-positive result on culture of a specimen of the aspiration fluid. Only two (6 per cent) of the thirty-four hips that had a positive result on culture of fluid from the initial aspiration had a true-positive result. No hip had a true-positive result on culture of fluid that had been aspirated preoperatively without also having clinical and radiographic evidence of infection. On the basis of these findings, we recommend that aspiration be performed in selected patients rather than routinely. It also should be performed only if a detailed clinical history suggests infection or if radiographs demonstrate focal lysis, aggressive non-focal lysis, or periostitis. PMID- 8419394 TI - The porous-coated anatomic total hip prosthesis, inserted without cement. Results after five to seven years in a prospective study. AB - The results of 100 primary arthroplasties with a porous-coated anatomic total hip prosthesis that were performed consecutively in ninety-one patients were followed prospectively for five to seven years after the operation or until death. Clinical Harris hip ratings and anteroposterior and lateral radiographs were made preoperatively, several times in the first postoperative year, and at annual intervals thereafter. Average sequential hip ratings were maintained between 92 and 93 points during the follow-up interval. Pain in the thigh, which did not limit function, was recorded at the yearly examinations from the first to the fifth year. After one year, 18 per cent of the thighs were painful, and in succeeding years, 19 per cent, 23 per cent, 26 per cent, and 15 per cent were painful. Radiographs demonstrated that the fixation of the femoral component was by ingrowth of bone in 94 per cent of the hips, by stable fibrous fixation in 1 per cent, and by unstable fibrous fixation in 5 per cent, according to the criteria of Engh et al. By five years, 6 per cent of the acetabular components had migrated, and 5 per cent of the femoral components had subsided. Two acetabular components had been revised, and one revision of a femoral component was pending. The evaluated device was an early-generation prosthesis that was designed for insertion without the use of cement. Further study is necessary to determine the long-term durability of the prosthesis-bone interface. PMID- 8419395 TI - Two cell lines from bone marrow that differ in terms of collagen synthesis, osteogenic characteristics, and matrix mineralization. AB - Two cloned cell lines were isolated from cultures of mouse bone-marrow cells. One of the lines, D1, exhibited osteogenic properties and synthesized type-I collagen (alpha 1)2 alpha 2. The second cell line, D2, was not osteogenic and produced a collagen homotrimer (alpha 1)3. Whereas the extracellular matrix of the D1 cell cultures contained striated collagen fibrils, presumably composed of type-I collagen, the homotrimer-producing D2 cells did not demonstrate striated collagen fibrils. Instead, they had thin filaments without detectable striations. Sodium ascorbate stimulated collagen synthesis at the transcriptional level in both the D1 and the D2 cells. The bone-producing characteristics of D1 in vitro included high levels of alkaline phosphatase, increased cyclic adenosine monophosphate on treatment with parathyroid hormone, and expression of osteocalcin mRNA. The D1 cells, unlike the D2 cells, produced a mineralized matrix in vitro. Mineralization in the cultures of the D1 cells occurred in nodules of increased cell density, which also contained the cells with the highest concentrations of collagen mRNA, as shown by in situ hybridization. When the D1 cells were implanted in a diffusion chamber in vivo, a mixture of both osteogenic and adipogenic tissues was formed. This indicates that the D1 cell line is derived from an early marrow stromal precursor that is multipotential. PMID- 8419396 TI - Inhibitory effects of a bacteria-derived sulfated polysaccharide against basic fibroblast growth factor-induced endothelial cell growth and chemotaxis. AB - The effects of sulfated polysaccharides on the growth and chemotaxis of endothelial cells promoted by basic fibroblast growth factor (bFGF), a heparin binding growth factor, and epidermal growth factor (EGF), a non-heparin-binding growth factor, were examined. The binding abilities of these two growth factors to D-gluco-D-galactan sulfate (DS-4152) were the same as to heparin. DS-4152 inhibited the growth and chemotaxis of the cells stimulated by bFGF, and prevented the binding of bFGF to the cells at both its low and high affinity binding sites: the former and the latter are heparin-like molecules and receptor proteins for bFGF, respectively. However, DS-4152 affected neither the binding of EGF to endothelial cells nor the proliferation and chemotaxis of the cells stimulated by the factor. Heparin also inhibited the binding of bFGF to low affinity binding sites to the same degree as DS-4152, but had little effect on the binding of bFGF to high affinity sites and no effects on bFGF-induced endothelial cell growth. Chondroitin sulfate A prevented neither the binding of bFGF to both sites of the cells nor bFGF-induced cell proliferation. We thus concluded that the inhibitory effects of DS-4152 against the growth and chemotaxis of endothelial cells induced by bFGF might be due to the prevention of bFGF binding to its receptor proteins resulting from the binding of DS-4152 to bFGF. PMID- 8419397 TI - Localization and quantitation of calcium pools and calcium binding sites in cultured human keratinocytes. AB - Calcium plays a crucial role in regulating the growth and differentiation of cultured keratinocytes. However, the mechanism(s) of this regulation is not clear. Prior studies have shown that intracellular free calcium (Cai) increases with keratinocyte differentiation. In this study, in order to evaluate the role of cytosolic free calcium and organelle-bound calcium in keratinocyte differentiation, we quantitated and localized calcium pools in keratinocytes, utilizing the fluorescence probe indo-1 and ion-capture cytochemistry, respectively. Cai of undifferentiated keratinocytes was 80-120 nM, whereas Cai of differentiated keratinocytes was 200-300 nM depending on the extent of differentiation. The Cai of individual cells in an undifferentiated colony was heterogeneous (60-160 nM) with larger cells displaying higher Cai. Heterogeneity also was observed in the intracellular calcium-containing precipitates in the different layers of stratifying keratinocyte cultures using the cytochemical technique. Calcium precipitates were abundant in the lower cell layers, progressively decreasing apically, with the uppermost layer devoid of precipitates. Calcium-containing precipitates appeared as fine-to-coarse electron dense granules on the plasma membrane, within the cytosol, mitochondria, nucleus, and vacuolar organelles. Whereas ionomycin in the presence of extracellular calcium increased the amount of intracellular calcium precipitates, EGTA removed calcium precipitates from organelles. Unlike intact epidermis, keratinocytes displayed no extracellular calcium reservoirs. Putative calcium binding sites, visualized by trivalent lanthanum (La) binding, were abundant on cell membranes and desmosomes of basaloid cells, but decreased in the upper cell layers. These studies revealed differences in the distribution of free ionic calcium (as determined by the fluorescence technique) and organelle-bound calcium (as determined by the cytochemical technique). Striking differences were also observed in calcium localization between intact epidermis and cultured epidermal cells. The localization pattern of calcium in cultured keratinocytes may reflect the hyperproliferative state of these cells, as in psoriatic epidermis, and/or the absence of a normal permeability barrier in these submerged cultures. PMID- 8419398 TI - Differential induction of stromelysin mRNA by bovine articular chondrocytes treated with interferon-gamma and interleukin-1 alpha. AB - Articular chondrocytes from rheumatoid joints have been shown to express class II major histocompatibility (MHC) antigens that were correlated with the presence of interferon-gamma (IFN-gamma) in the inflamed joint. Chondrocytes expressing MHC antigens function as antigen presenting cells and thus stimulate lymphocyte proliferation. These responses suggest a powerful role for the IFN-gamma stimulation of chondrocytes. The present studies were designed to examine the functional role of chondrocytes exposed to IFN-gamma during cartilage degradation that occurs in synovial disease. Destruction of cartilage in arthritis is partially attributable to metalloproteinases released by the chondrocytes in response to interleukin-1 (IL-1). Bovine articular chondrocytes treated with interleukin-1 alpha (IL-1 alpha) produced enhanced levels of stromelysin mRNA, however, Northern blots could not determine the percentage of cells responding. Exposure of bovine articular chondrocytes to IFN-gamma induced the expression of bovine HLA-DR (boHLA-DR) antigen in 50% of the cells. Using a modified cell sorting technique, chondrocytes that expressed class II MHC antigens produced two fold greater stromelysin mRNA than chondrocytes that did not express this antigen. In contrast, collagen type II mRNA levels were similar in chondrocytes, regardless of the expression of class II MHC antigens. In situ hybridization studies showed that less than half of all cartilage chondrocytes were induced to synthesize stromelysin mRNA. These observations suggest that IFN-gamma stimulates specific subpopulations of chondrocytes to be functionally active in inflammation induced metalloprotease secretion. PMID- 8419399 TI - Increase in epidermal growth factor receptor and its messenger ribonucleic acid levels with differentiation of human trophoblast cells in culture. AB - Epidermal growth factor receptor (EGFR) expression was studied during the differentiation of human trophoblast cells in culture. In vitro, intravillous mononuclear cytotrophoblasts aggregate and fuse within 24 h to form a syncytium. This morphological differentiation was associated with a significant twofold increase in specific 125I-EGF binding capacity (P < 0.01). Scatchard analyses showed an apparent rise in the number of high-affinity binding sites (0.33 +/- 0.04 and 0.63 +/- 0.07 pmol/mg protein at 24 and 48 h, respectively), with no change in their affinity (1.34 and 1.42 x 10(-10) mol/L). Affinity labeling of 125I-EGF in cultured trophoblast cells followed by SDS-PAGE and autoradiography revealed a band of 175 KDa corresponding to EGFR, the intensity of which increased with the time in culture. EGF-dependent phosphorylation of membrane proteins from cultured trophoblast cells revealed major phosphorylated proteins of 170 KDa (EGFR) and 35 KDa, which were both increased at 48 h, indicating a rise in EGFR-kinase activity during syncytium formation. Northern blot analysis of EGFR-mRNA, followed by hybridization with a 32P-cDNA probe for EGFR, revealed an increase in EGFR gene expression in syncytiotrophoblasts, as compared to cytotrophoblasts. Thus, the increase in bioactive EGFR observed during the differentiation of trophoblast cells was due to an increase in their synthesis. Cultured trophoblast cells are therefore a good model of spontaneous up regulation of EGFR expression with cell differentiation. PMID- 8419400 TI - Pulsatile and steady flow induces c-fos expression in human endothelial cells. AB - The effects of pulsatile and steady fluid flow on the mRNA levels of proto oncogenes c-fos, c-jun, and c-myc in cultured human umbilical vein endothelial cells (HUVEC) were investigated. c-fos mRNA levels in stationary cultures were very low. A 1 Hz pulsatile flow with an average shear stress of 16 dynes/cm2 induced a dramatic increase of c-fos mRNA levels in HUVEC 0.5 h after the onset of flow, which declined rapidly to basal levels within 1 h. Steady flow with a similar shear stress also induced a transient increase of c-fos mRNA levels, but to a lesser extent. In addition, increased c-fos mRNA levels were observed when low shear (2-6 dynes/cm2) was replaced by high shear (16-33 dynes/cm2). Pulsatile and steady flow caused a slight increase of c-jun and c-myc mRNA levels. The role of pulsatility was also investigated in platelet-derived growth factor (PDGF) expression. Pulsatile flow induced a transient increase of PDGF A- and B-chain mRNA levels with peaks at 1.5-2 h. Pulsatile flow, which was more stimulatory in mediating c-fos expression, however, was less stimulatory than steady flow in mediating PDGF expression. By using various inhibitors, protein kinase C was found to be an important mediator in flow-induced c-fos expression, with the involvement of G proteins, phospholipase C, and intracellular calcium. Protein kinase C was previously shown as a possible major mediator in flow-induced PDGF expression which, at least partly, appeared to follow the induction mechanism of c-fos, suggesting a possible connection between c-fos and PDGF induction. However, the c-fos antisense treatment, which significantly inhibited c-fos transcription, failed to block the flow-induced PDGF expression, suggesting that flow-induced c-fos expression may not play an important role in the mechanism of flow-induced PDGF expression. The difference in the induction of c-fos and PDGF expression under pulsatile as compared to steady flow indicates that a complex, flow-mediated regulatory mechanism of gene expression exists in HUVEC. The increased expression of these proto-oncogenes mediated by flow may be important in regulating long-term cellular responses. PMID- 8419401 TI - Internalization of basic fibroblast growth factor (bFGF) in cultured endothelial cells: role of the low affinity heparin-like bFGF receptors. AB - We have shown (Presta et al., Cell Regul., 2:719-726, 1991) that a long-lasting interaction of basic fibroblast growth factor (bFGF) with endothelial GM 7373 cells is required to induce cell proliferation. In the present work, we have investigated the interaction of 125I-bFGF with GM 7373 cells, its pathway of internalization, and its intracellular fate under the same experimental conditions previously utilized to assess the mitogenic activity of the growth factor. Cell cultures were incubated with 10 ng/ml 125I-bFGF for 2 h at 4 degrees C. Then, cells were shifted to 37 degrees C without changing the medium. A rapid down-regulation of high affinity sites, paralleled by a rapid internalization of 125I-bFGF, was observed during the first 1-2 h at 37 degrees C. After 6-8 h, also low affinity sites down-regulate. This was paralleled by a continuous internalization of 125I-bFGF and by a slow disappearance of the growth factor from the culture medium. This suggests that GM 7373 cells activate, when exposed to bFGF for a long period of time, a late internalization pathway mediated by low affinity sites. This hypothesis was supported by the following experimental evidence: 1) soluble heparin inhibited the prolonged internalization of 125I-bFGF and its binding to low affinity sites with the same potency; 2) treatment of GM 7373 cells with heparinase, which removes most of the low affinity sites, also inhibited the prolonged internalization of 125I-bFGF. 125I-bFGF internalized via low affinity sites was partially protected from lysosomal degradation. This was the case also when 125I-bFGF was internalized in the presence of soluble heparin, suggesting that the complexes bFGF-cell surface glycosaminoglycan and bFGF soluble heparin are maintained during the internalization of the growth factor. Moreover, the capacity of soluble heparin to inhibit the mitogenic activity of bFGF also when added to cell cultures several hours after the growth factor indicates that the requirement for a prolonged interaction of bFGF with GM 7373 cells in order to induce cell proliferation might be related to the late internalization of the growth factor via low affinity sites. PMID- 8419402 TI - Cell cycle regulated expression of nucleolar antigen P120 in normal and transformed human fibroblasts. AB - Normal and SV40 virus-transformed WI-38 human lung fibroblasts were serum starved and refed, or synchronized by double thymidine block and released from the block. At different time points in the cell cycle, steady state levels of P120 mRNA and P120 protein content of the cells were determined by densitometric scans of Northern and Western blots. At the same time points, [3H]thymidine uptake was measured and flow cytometric analysis performed for DNA content and P120 antigen staining. Levels of P120 protein and P120 mRNA were approximately 4 times greater in non-synchronous, exponentially growing transformed cells than in similarly growing normal cells. Early G1-phase cells, synchronized either with serum deprivation or with metabolic block, contained only a trace amount of P120 protein and mRNA. The P120 gene was transcribed early in G1 and P120 protein synthesis initiated in middle G1. A dramatic increase of P120 protein level occurred in S-phase with a corresponding mRNA peak preceding the P120 protein peak. These results indicate that P120 is overexpressed in transformed WI-38 cells and that P120 is temporally regulated during the cell cycle of both transformed and normal fibroblasts. The dramatic increase in P120 protein expression at the G1 to S boundary suggests that P120 may play a role in the regulation of cell cycle and increased nucleolar activity that is associated with cell proliferation. PMID- 8419403 TI - Major influence of cell differentiation status on characteristics of proteoglycans synthesized by cultured rabbit renal proximal tubule cells: role of insulin and dexamethasone. AB - To analyze the influence of epithelial cell differentiation and the effects of hormones on the characteristics of cell-associated and secreted proteoglycans (PGs), we studied their distribution, synthesis, and biochemical features in a model of renal proximal tubule cells in primary culture in which cell differentiation could be controlled by medium composition. In cells cultured in serum-free, hormonally defined medium supplemented with insulin and dexamethasone that exhibited a high degree of morphological and functional proximal differentiation (Ronco et al., 1990), cell-associated PGs were similar to those extracted in vivo by their size estimated by Sepharose CL-6B chromatography (Kav = 0.27, vs. 0.26), composition (heparan-sulfate), and localization in a continuous basal layer of extra-cellular matrix (ECM). In contrast, major quantitative and qualitative anomalies of cell-associated PGs were observed in poorly differentiated cells grown in 1% fetal calf serum-supplemented medium (FCS). PGs alterations included: (1) reduced and irregular expression of PGs at the cell basal pole, (2) a 2.8-fold decrease in [35S]-sulfate incorporation into cell-associated PGs, (3) a 3.1-fold increase in trypsin-releasable PGs, and (4) the emergence of a high MW PG composed exclusively of chondroitin-sulfate (CS) (Kav = 0.09 on Sepharose CL-6B) as well as of putative free CS-glycosaminoglycan (GAG) chains (Kav = 0.49 on Sepharose CL-6B). The same alterations were identified in the basal defined medium devoid of hormones but were partially or totally abolished by addition of insulin and dexamethasone, respectively. At variance with cell-associated PGs, production and biochemical features of secreted PGs were not influenced by cell differentiation status and medium composition. PMID- 8419404 TI - 19F NMR studies of changes in membrane potential and intracellular volume during dexamethasone-induced apoptosis in human leukemic cell lines. AB - The induction of apoptosis in leukemic cells by dexamethasone is well known, but the mechanism of this type of cell death and of dexamethasone resistance by some variants is still poorly understood. Apoptotic cell death is preceded by many changes in cellular properties, such as glucose metabolism, cell size, cell density, and others. In this study, 19F-NMR has been used to characterize changes in cell membrane potential and intracellular accessible volume during dexamethasone induced apoptosis. One dex-sensitive (CEM-C7) and three dex resistant variants (CEM-C1, CEM-ICR27, and CEM-4R4) were examined. We have observed separate intracellular and extracellular resonances for trifluoroacetate and trifluoroacetamide added to suspended leukemic cells. From the equilibrium distribution of these fluoro-compounds between intra and extracellular spaces, the changes in membrane potential and intracellular accessible volume were calculated. The membrane potential for CEM-C7 cells was found to significantly decrease in the presence of dexamethasone (9-mV decrease within 18 h of dexamethasone treatment), while that of CEM-ICR27 was found in some samples to increase on dexamethasone incubation. The membrane potential for CEM-C1 decreased slightly, while that of CEM-4R4 was not appreciably affected by dexamethasone. The reduction of membrane potential seems to be an early step in the mechanism of dexamethasone induced apoptosis. Although the intracellular volume varied with cell type and dexamethasone incubation (for CEM-C7), the fractional intracellular volume (alpha = Vin/Vcell) was found to be the same (0.82 +/- 0.06) for all the cell lines in the presence and absence of dexamethasone. PMID- 8419405 TI - Transduced endothelial cells expressing high levels of tissue plasminogen activator have an unaltered phenotype in vitro. AB - Elevated cellular plasminogen activator activity has been associated with significant alterations in the in vitro phenotype of both malignant cell lines and nonmalignant endothelial cells. To examine the role of elevated cellular plasminogen activator activity in the production of altered endothelial cell behavior, bovine coronary artery endothelial cells were transduced with a retroviral vector expressing large amounts of tissue plasminogen activator. Cells transduced with the tissue plasminogen activator vector were compared with both untransduced cells and cells transduced with a control vector in a series of in vitro assays of cellular attachment, proliferation, migration, and invasion. The morphology of the 2 transduced populations was unchanged. There was a small decrease (5-15%) in the horizontal migration rate of both transduced cell populations, as compared with that of untransduced cells. No significant differences were detected among the three cell populations in any of the other assays. We conclude that expression of high levels of tissue plasminogen activator does not specifically affect endothelial cell phenotype in vitro. These data lend support to the hypothesis that elevated plasminogen activator activity is necessary but not sufficient to produce alterations in endothelial cell behavior. PMID- 8419406 TI - Contrasting effect of transforming growth factor type beta 1 (TGF-beta 1) on proliferation and interleukin-2 receptor expression in activated and rapidly cycling immature (CD3-CD4-CD8-) thymocytes. AB - Transforming growth factor beta (TGF-beta) is a cytokine with immunoregulatory properties that acts negatively on T lymphocyte proliferation. However, with the EL 4-6.1 variant of the murine thymoma EL 4 activated with phorbol ester and/or interleukin-1 (IL-1), we recently found that it up-regulates interleukin-2 receptor (IL-2R) expression. Since EL 4-6.1 cells share phenotypic and functional characteristics with the immature thymic subset lacking CD4 and CD8 accessory molecules (DN), we investigated the effect of TGF-beta 1 on the IL-2R 55kD alpha chain expression and proliferation of activated DN cells and especially in DN cells that do not express CD3. We observed that TGF-beta 1 was able to increase both the percentage of CD3-DN cells expressing IL-2R alpha chains and the expression of IL-2R alpha chain in these cells. This stimulatory effect of TGF beta 1 was distal from early transduction events. In addition, TGF-beta 1 was found to modulate CD3-DN cell proliferation. During differentiation in the thymus, CD3-DN cells transiently express the IL-2R alpha chain of the IL-2R and these IL-2R+ CD3-DN cells are preprogrammed to down-regulate the IL-2R alpha chain and up-regulate the CD4 and CD8 accessory molecule. We thus also tested the effect of TGF-beta 1 on IL-2R alpha chain expression in these in vitro differentiating CD3-DN cells. We found that TGF-beta 1 neither significantly affected IL-2R expression nor changed CD4 or CD8 expression. Hence, in CD3-DN cells, the effect of TGF-beta 1 on IL-2R expression seems to be restricted to proliferating cells. PMID- 8419407 TI - Differential effects of SPARC and cationic SPARC peptides on DNA synthesis by endothelial cells and fibroblasts. AB - SPARC (secreted protein, acidic and rich in cysteine), also known as osteonectin, is an extracellular Ca+2-binding glycoprotein that inhibits the incorporation of [3H]-thymidine and delays the onset of S-phase in synchronized cultures of bovine aortic endothelial (BAE) cells. This effect appears not to be dependent on the functional properties of SPARC associated with changes in cell shape or inhibition of cell spreading. In this study we investigate the conditions under which cell cycle modulation occurs in different types of cells. Human umbilical vein endothelial cells, a transformed fetal BAE cell line, and bovine capillary endothelial cells exhibited a sensitivity to SPARC and a cationic peptide from a non-Ca+2-binding region of SPARC (peptide 2.1, 0.2-0.8 mM) similar to that observed in BAE cells. In contrast, human foreskin fibroblasts and fetal bovine ligament fibroblasts exhibited an increase in the incorporation of [3H]-thymidine in the presence of 25 microM-0.2 mM peptide 2.1; inhibition was observed at concentrations in excess of 0.4 mM. This biphasic modulation could be further localized to a sequence of 10 amino acids comprising the N-terminal half of peptide 2.1. A synthetic peptide from another cationic region of SPARC (peptide 2.3) increased [3H]-thymidine incorporation by BAE cells and fibroblasts in a dose-dependent manner. In endothelial cells, a stimulation of 50% was observed at a concentration of 0.01 mM; fibroblasts required approximately 100-fold more peptide 2.3 for levels of stimulation comparable to those obtained in endothelial cells. The observation that SPARC and unique SPARC peptides can differentially influence the growth of fibroblasts and endothelial cells in a concentration dependent manner suggests that SPARC might regulate proliferation of specific cells during wound repair and remodeling. PMID- 8419408 TI - Rapamycin blocks cell cycle progression of activated T cells prior to events characteristic of the middle to late G1 phase of the cycle. AB - The effects of rapamycin (RAP) on cell cycle progression of human T cells stimulated with PHA were examined. Cell cycle analysis showed that the RNA content of cells stimulated with PHA in the presence of RAP was similar to that of control T cells stimulated with PHA for 12-24 hr in the absence of the drug. This level was substantially higher than that seen in cells stimulated in the presence of cyclosporin A (CsA), an immunosuppressant known to block cell cycle progression at an early point in the cycle. However, the point in the cell cycle at which RAP acted appeared to be well before the G1/S transition, which occurs about 30-36 hr after stimulation with PHA. In an attempt to further localize the point in the cell cycle where arrest occurred, a set of key regulatory events leading to the G1/S boundary were examined, including p110Rb phosphorylation, which occurred at least 6 hr prior to DNA synthesis, p34cdc2 synthesis, and cyclin A synthesis. In control cultures, p110Rb phosphorylation was detected within 24 hr of PHA stimulation; p34cdc2 and cyclin A synthesis were detected within 30 hr. Addition of RAP to the cultures inhibited each of these events. In contrast, early events, including c-fos, IL-2, and IL-4 mRNAs expression, and IL 2 receptor (p55) expression, were only marginally affected, if at all, in PHA stimulated T cells. Furthermore, the inhibition of cell proliferation by RAP could not be overcome by addition of exogenous IL-2. These results indicate that RAP blocks cell cycle progression of activated T cells after IL-2/IL-2 receptor interaction but prior to p110Rb phosphorylation and other key regulatory events signaling G1/S transition. PMID- 8419409 TI - Different relationships between cellular adenosine or 3'-deoxyadenosine phosphorylation and cellular adenine ribonucleotide catabolism may be obtained. AB - Treatment of BALB/c-3T3 mouse fibroblasts with 3'-deoxyadenosine led to a rapid accumulation of 3'-deoxyadenosine phosphates and the kinetics of this process has been determined. Concomitant with accumulation of these compounds, the adenine ribonucleotide pool was reduced. The kinetics of the two processes suggested that they were tightly coupled. The inhibitory effect of relatively high concentrations of coformycin indicated that IMP was an intermediate in the catabolic pathway. Similar experiments with Ehrlich ascites tumor cells were performed in Ringer-Hepes solution at pH 6.5 or 7.5 and with varying concentrations of orthophosphate. The experiments were performed with cells where ATP was [3H]-labeled. This allowed the determination of the catabolism of adenine ribonucleotides to labeled nucleosides under conditions where added adenosine was phosphorylated. The results showed that at low phosphate concentration (5.8 mM) at pH 6.5 adenosine may be phosphorylated at a rate that was completely balanced to the concomitant catabolism of adenine ribonucleotides; that is, there was apparently a tight kinetic coupling between anabolism of adenosine and catabolism of adenine ribonucleotides. With 3'-deoxyadenosine a corresponding effect was obtained although the apparent coupling between phosphorylation of 3' deoxyadenosine and catabolism of adenine ribonucleotides was not complete. When experiments were performed at the same pH but at high concentration of phosphate (45 mM) there was in contrast no coupling between the two processes; that is, ATP was present in constant amounts while 3'-deoxyadenosine phosphates accumulated at a high rate. In experiments with adenosine under these conditions there was still some although a relatively limited degree of apparent coupling between phosphorylation of adenosine and catabolism of adenine ribonucleotides. In both lines of cells used and with both adenosine and 3'-deoxyadenosine, the main products of the catabolism of adenine ribonucleotides were inosine and hypoxanthine. With 3'-deoxyadenosine there was in addition (about 20%) formation of xanthosine, suggesting that IMP dehydrogenase had also been activated. These results lead to the suggestion that adenosine (or 3'-deoxyadenosine) may be phosphorylated in two ways. 1) Phosphorylation may depend on an adenosine kinase unrelated to catabolism of adenine ribonucleotides. 2) Phosphorylation may be tightly coupled to catabolism of adenine ribonucleotides. A nucleoside phosphotransferase may catalyze the transfer of a phosphoryl group from IMP to adenosine (or 3'-deoxyadenosine) to form AMP (or 3'-dAMP) and inosine, a process that may be tightly coupled to an AMP deaminase reaction. The IMP formed in the latter reaction may not be released but transferred to the phosphotransferase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419410 TI - Growth inhibitory activity in extracts from human fibroblasts. AB - Indirect evidence for the presence of a growth inhibitor in normal human fibroblasts has been obtained previously; the inhibitory activity has been found associated with crude cell extracts, but the molecule responsible for the growth inhibition has never been isolated. We have isolated a glycopeptide fraction from human fibroblast cultures, whose synthesis decreases when the cells are stimulated into the division cycle. It was separated by electric charge, lectin affinity, and molecular mass. When added to quiescent cells simultaneously with a growth stimulus, the glycopeptide reduces DNA synthesis activity. The relationship of the kinetics of the synthesis of the glycopeptide with the cell division cycle and its molecular weight are different from what has been described so far for other growth regulators. The decreased synthesis of this inhibitor, induced by growth factors, seems to be one of the requirements for the initiation of the division cycle by human fibroblasts. This response to growth factors was stable during the lifespan of the fibroblast population and became less pronounced only in cells at the end of their replicative potential. PMID- 8419411 TI - The survivor. PMID- 8419412 TI - Chest pain in a young man with chronic abdominal bloating. PMID- 8419413 TI - Tumor suppressor genes. PMID- 8419414 TI - Unstable angina in a man with joint pain. PMID- 8419415 TI - Immunopathogenesis of pulmonary tuberculosis. AB - Antimicrobial therapy quickly eradicates susceptible bacilli, but the fight against drug-resistant disease must be waged entirely by host defenses. Knowledge of the two main types of immune response against tuberculosis and of how to manipulate those mechanisms--leading to precisely designed recombinant BCG vaccines--is essential to mounting an effective attack on the current epidemic. PMID- 8419416 TI - MRI of injury to the lateral collateral ligamentous complex of the ankle. AB - We retrospectively evaluated the lateral collateral ligamentous complex of 43 patients who had complained of ankle pain following ankle sprain. The MR signs of ligamentous abnormality included discontinuity or absence, increased signal within the ligament, and ligamentous irregularity or waviness with normal thickness and signal intensity. Using these criteria, 30 anterior talofibular, 20 calcaneofibular, and no posterior talofibular ligament injuries were diagnosed. Compared with surgery (nine patients), MRI demonstrated six of seven anterior talofibular ligament injuries and six of six calcaneofibular ligament injuries. Magnetic resonance showed ligamentous abnormalities in 12 of 23 cases with normal stress radiography. Magnetic resonance imaging provides useful information for the evaluation of patients presenting with chronic pain after ankle sprain. PMID- 8419417 TI - Noninvasive CT determination of arterial blood concentration of meglumine iothalamate. AB - A quantity that often must be determined in physiological imaging studies is the blood concentration of the tracer over time. This is usually performed by direct arterial or venous blood sampling. We studied the relationship between the concentration of meglumine iothalamate in arterial blood and values determined from voxels containing large blood vessels in a series of CT images at the same location over time. After correction for volume averaging based on a single venous blood sample, there was an excellent correlation between the two blood curves. Differences between the curves were shown to be inconsequential by a simulation of transcapillary transport determinations. We thus conclude that determination of plasma concentration from CT images is a reliable technique for CT transcapillary transport studies. PMID- 8419418 TI - [18F]fluorodeoxyglucose uptake in tumors: kinetic vs. steady-state methods with reference to plasma insulin. AB - [18F]Fluorodeoxyglucose (FDG) uptake in noncerebral tumors is commonly reported as tissue radioactivity concentration normalized to injected dose and body weight. We studied the feasibility of this approach by imaging 68 tumors in 46 oncologic patients with dynamic FDG-PET and compared kinetic and static methods of quantitation of FDG uptake. Further, the effect of plasma glucose and insulin concentration on the obtained quantitative indexes was analyzed in all patients. The metabolic rate for FDG was strongly associated with normalized uptake value adjusted for injected dose (r = 0.92, p < 0.0001), dose and patient weight (r = 0.91, p < 0.0001), and dose and body surface area (r = 0.94, p < 0.0001). The FDG uptake was not related to plasma glucose concentration under euglycemic (< or = 6.5 mmol/L) conditions, but was low in two diabetic patients with overt hyperglycemia. Hyperinsulinemia was associated with a low to moderate FDG uptake, probably exerting its action through a metabolic shift of tracer influx to muscle and fat. Our results show that a single scan in the steady-state phase, e.g., 45 60 min from the injection, can be used for assessment of FDG uptake in tumors, making frequent blood sampling during imaging unnecessary. However, glucose concentration in blood must be monitored in patients with known or suspected abnormalities in glucose metabolism. PMID- 8419419 TI - Postradiation cerebellar necrosis mimicking tumor: MR appearance. AB - Cerebellar necrosis associated with therapy of primary gliomas is unusual. We present the MR findings in a patient with postradiation multifocal cerebellar necrosis mimicking tumor dissemination. PMID- 8419420 TI - MR features of tuberculous arachnoiditis. AB - Although tuberculosis is an uncommon cause of spinal arachnoiditis, it needs to be differentiated from other causes of arachnoiditis because it is a treatable disease. Myelography, which is the imaging modality of choice for the chronic adhesive stage of tuberculous arachnoiditis, usually reveals irregularity of the thecal sac, nodularity and thickening of nerve roots, clumping of roots to each other or to the thecal sac, or CSF block. Recently Gd-diethylenetriamine pentaacetic acid enhanced MRI has been found to be useful for the active phase of tuberculous infection, effectively demonstrating abnormally thickened and enhancing meninges, intra- and extramedullary tuberculous nodules, and spinal cord changes. We present the MRI features in a case of acute tuberculous arachnoiditis. PMID- 8419421 TI - Cervical lymphadenopathy due to Kikuchi disease: US and CT appearance. AB - A patient with Kikuchi disease (histiocytic necrotizing lymphadenitis), who came to clinical attention because of cervical lymphadenopathy, is presented. CT and ultrasonographic findings of this disease are described. The importance of this entity lies in the fact that it can be easily confused clinically, pathologically, and radiologically with malignant lymphadenopathy, especially lymphoma. PMID- 8419422 TI - Fluid-fluid levels in a unicameral bone cyst: CT and MR findings. AB - We present a case of unicameral bone cyst with fluid-fluid levels evaluated with CT and MRI. A literature review revealed that fluid levels within aneurysmal bone cysts, giant cell tumors, chondroblastomas, and tel-angiectatic osteosarcomas have been described. PMID- 8419423 TI - Chronic bursitis presenting as a mass in the pes anserine bursa: MR diagnosis. AB - A case of chronic, nonspecific synovial inflammation presenting as a mass in the pes anserine bursa is demonstrated by MRI. The lesion was well demarcated and surrounded by a low intensity rim. On spin echo images it was homogeneous and of intermediate signal intensity, whereas on T2-weighted images it showed scattered areas of high signal producing a heterogeneous pattern. It is contrasted with a typical example of acute pes anserine bursitis, presenting as simple fluid within the bursa. The differential considerations vary accordingly between the acute and chronic forms of pes anserine bursitis, with the latter requiring differentiation from other forms of synovitis, synovial hemangioma, and synovial sarcoma. The MR features of these entities are reviewed as an aid in differential diagnosis. PMID- 8419424 TI - CT appearance of the distended trochanteric bursa. AB - The CT appearance of the distended trochanteric bursa noted in an asymptomatic patient is reported. The bursal distension was noted incidentally in a patient undergoing staging for renal cell carcinoma. On CT the distended bursa was noted as a septated low attenuation lesion at the site of insertion of the gluteus medius and minimus muscles on the greater trochanter of the femur. The lesion was not associated with any degenerative changes in the hip or greater trochanter. Distension of the trochanteric bursa may occur in asymptomatic patients and the appearance of the distended bursa must be recognized on imaging studies to avoid confusing it with other lesions. PMID- 8419425 TI - Value of CT in diagnosing nonneoplastic osteolysis in Paget disease. AB - We present an unusual case of osteolysis, likely due to a stress fracture, in a pagetic pubic bone simulating sarcomatous transformation. PMID- 8419426 TI - Parosteal ossifying lipoma: CT and MR findings. AB - Radiologic and imaging findings of parosteal ossifying lipoma along with pathologic correlation are presented in this case report. PMID- 8419427 TI - Intracranial MRA: single volume vs. multiple thin slab 3D time-of-flight acquisition. AB - Single volume three-dimensional (3D) time-of-flight (TOF) MR angiography is the most commonly used noninvasive method for evaluating the intracranial vasculature. The sensitivity of this technique to signal loss from flow saturation limits its utility. A recently developed multislab 3D TOF technique, MOTSA, is less affected by flow saturation and would therefore be expected to yield improved vessel visualization. To study this hypothesis, intracranial MR angiograms were obtained on 10 volunteers using three techniques: MOTSA, single volume 3D TOF using a standard 4.9 ms TE (3D TOFA), and single volume 3D TOF using a 6.8 ms TE (3D TOFB). All three sets of axial source images and maximum intensity projection (MIP) images were reviewed. Each exam was evaluated for the number of intracranial vessels visualized. A total of 502 vessel segments were studied with each technique. With use of the MIP images, 86% of selected vessels were visualized with MOTSA, 64% with 3D TOFA (TE = 4.9 ms), and 67% with TOFB (TE = 6.8 ms). Similarly, with the axial source images, 91% of selected vessels were visualized with MOTSA, 77% with 3D TOFA (TE = 4.9 ms), and 82% with 3D TOFB (TE = 6.8 ms). There is improved visualization of selected intracranial vessels in normal volunteers with MOTSA as compared with single volume 3D TOF. These improvements are believed to be primarily a result of decreased sensitivity to flow saturation seen with the MOTSA technique. No difference in overall vessel visualization was noted for the two single volume 3D TOF techniques. PMID- 8419428 TI - Monoarticular gout following trauma: MR appearance. AB - We report a case of gout with monoarticular tophaceous involvement of the proximal interphalangeal joint of the middle finger, emphasizing MR findings. To the best of our knowledge, the MR appearance of gout is not commonly known. PMID- 8419429 TI - Infrarenal vena caval injury following blunt trauma: CT findings. AB - Isolated infrarenal vena caval injury following blunt trauma is rare. In most patients, poor clinical status usually precludes imaging prior to laparotomy. When CT is performed, however, signs that allow a diagnosis of caval injury to be made are (a) retroperitoneal hematoma with paracaval epicenter; (b) irregular vena caval contour; and (c) extravasation of contrast-enhanced blood from the cava. PMID- 8419430 TI - MRI of vertebral sarcoidosis. PMID- 8419431 TI - The "cluster" sign in macronodular hepatic tuberculosis: CT features. PMID- 8419432 TI - Renal angiomyolipoma extending into inferior vena cava: MR demonstration. PMID- 8419433 TI - Evaluation of nondistended pyriform sinus without respiratory maneuvers. AB - Detection of a nondistended pyriform sinus on cross-sectional imaging studies represents a diagnostic dilemma. The finding may be an inconstant physiologic phenomenon without clinical significance, or it may be due to tissue thickening and lack of pliability related to neoplasia or inflammation. Rescanning during respiratory maneuvers may clarify the anatomy, but full patient cooperation is needed. We demonstrate a method (turning the patient's head away from the side of the nondistended sinus) that induces distention of the pyriform sinus but does not require active patient participation. PMID- 8419434 TI - Persistent trigeminal artery detected with standard MRI. AB - The trigeminal artery is the most common of the primitive carotid-basilar anastomoses to persist into adulthood. Prior to the introduction of MRI, the diagnosis of persistent trigeminal artery (PTA) could only be made at cerebral angiography. This study compares angiography and standard brain MRI in 11 cases of PTA. Nine of the 11 PTAs were identified with MRI. Axial imaging gave the best definition of the course of the PTA. PMID- 8419435 TI - 3D MRI of the cervical spine: low flip angle FISP vs. Gd-DTPA TurboFLASH in degenerative disk disease. AB - The authors undertook this study to compare bright and dark CSF three-dimensional (3D) gradient-echo (GE) MR techniques to answer the following questions: Could a single Gd-DTPA enhanced T1-weighted GE volume sequence (with multiplanar reformats) be diagnostically equivalent for degenerative cervical disk disease to a standard sequence consisting of sagittal T1-weighted spin echo and axial low flip angle volume GE images (with reformatted images)? Does performing oblique coronal reformats perpendicular to the course of exiting cervical nerve roots improve diagnostic confidence over axial images alone? Thirty-one consecutive patients received a "routine" MR examination consisting of a sagittal T1-weighted spin echo and axial low flip angle volume sequence (FISP) [(35/7/5), 64 slices, 2 mm slice thickness, 192 x 256 matrix, 7.2 min]. Each patient was then given 0.1 mmol/kg Gd-DTPA intravenously, and reimaged with a T1-weighted volume GE sequence [(13/6/12), acquired as 128-1.2 mm coronal partitions, 192 x 256 matrix, 5.5 min]. Sequences were reconstructed on the standard diagnostic console in 1 mm increments. Sets of examinations (routine vs T1-weighted volume) were independently interpreted by three neuroradiologists for location, type, and severity of extradural degenerative disease. There was no strong or consistent trend for increased detection of disease by one imaging sequence over the other. For lateral disk disease, only 3% of the observations were in discordance. For disk disease, there was close agreement in the severity scores. All readers indicated that additional information was provided by the reformatted images more frequently with TurboFLASH (fast low angle shot) than with FISP. All readers indicated that increased confidence was provided by the reformatted images more frequently with TurboFLASH than with FISP. A single 3D contrast-enhanced TurboFLASH sequence is diagnostically equivalent to a set of two-dimensional T1 weighted sagittal spin echo and 3D axial low flip angle sequences for assessing the location and degree of cervical extradural degenerative disease. A screening examination of the cervical spine could be performed with a single contrast enhanced 5.2 min study, and then relying on computer postprocessing to provide additional imaging planes. PMID- 8419436 TI - Whole body skeletal imaging with [18F]fluoride ion and PET. AB - Using our recently reported whole body PET imaging technique, we performed whole body PET studies of the skeletal system with [18F]fluoride ion in 19 patients with a range of malignant and benign skeletal conditions and in 19 normal male volunteers. The technique produces two-dimensional projection images of the entire skeletal system ("a PET bone scan"), in addition to coronal, sagittal, and axial tomographic images of the skeletal system. The tomographic images had a 13% higher lesion detection sensitivity than the projection images. Whole body PET skeletal imaging with [18F]fluoride ion is technically feasible, provides images of excellent quality, and may be coupled with more quantitatively precise kinetic PET [18F]fluoride ion studies (over limited regions of the body) when numerical estimates of skeletal [18F]fluoride ion uptake are desired. The method is potentially useful in clinical applications where the high resolution and numerical precision of PET are of particular value (e.g., in accurately defining the anatomic location and extent of lesions and in assessing changes in bone metabolism on serial studies). PMID- 8419437 TI - A pitfall in ultrafast CT scanning for the detection of left atrial thrombi. AB - We studied 41 patients with mitral stenosis by ultrafast CT (UFCT) and transesophageal echocardiography to detect left atrial thrombi. Cardiac UFCT was performed twice after contrast medium injection to obtain early (during injection lasting 40-60 s) and late (approximately 5 min after beginning injection) phase images. There were 10 patients (24%) in whom a filling defect detected in the early phase disappeared in the late phase. The site of filling defects was the left atrial appendage in nine patients and the left atrium in one patient. All of the filling defects were in the ventral side of the left atrium. Furthermore, all of those patients had chronic atrial fibrillation. Transesophageal echocardiography revealed no thrombus in the area of the filling defect in the early phase. We believe that blood stasis existed in those patients. This finding leads to a false-positive result when only early phase images are obtained. The diagnosis of thrombi should be made only when a filling defect is observed in both phases. Late phase scanning is necessary in the diagnosis of left atrial thrombi. PMID- 8419438 TI - High resolution CT in respiratory bronchiolitis-associated interstitial lung disease. AB - High-resolution CT findings have been described for several diffuse lung diseases. Respiratory bronchiolitis-associated interstitial lung disease (RB-ILD) is an inflammatory lung disorder associated with cigarette smoking. This condition has only recently been described and distinguished from desquamative interstitial pneumonitis, which it closely resembles. We describe high-resolution CT findings in five cases of biopsy-proven RB-ILD. The findings are variable and range from no detectable abnormality to atelectasis, ground-glass opacities, emphysema, and linear and reticular interstitial abnormalities. PMID- 8419439 TI - CT and MR features of primary pulmonary hemangiopericytomas. AB - We investigated the radiological findings of four patients with malignant pulmonary hemangiopericytomas, comparing plain chest radiography, CT, and MR. The radiographic presentation ranged from a small single lesion to a tumor almost completely occupying a hemithorax. Pathognomonic radiological features of malignant pulmonary hemangiopericytomas could not be established, but characteristic features include a large lobulated well-circumscribed mass with an encapsulated appearance, contrast enhancement of the margin, and no signs of surrounding compression atelectasis. The tumors were associated with late onset or a paucity of chest symptoms. Additional information regarding heterogeneity and effects on adjacent structures was provided by MR and was helpful in suggesting the diagnosis of malignant pulmonary hemangiopericytoma. PMID- 8419440 TI - Occult pneumothorax in patients with abdominal trauma: CT studies. AB - Abdominal CT, which routinely includes the lower thorax, is an important adjunct to supine chest radiography in detecting chest injury in patients with blunt abdominal trauma. In 1,086 consecutive patients with blunt abdominal trauma, 223 of whom had both supine chest radiography and abdominal CT, 49 patients examined with both techniques had pneumothoraces, 28 of them occult (seen only on CT). To help guide management, we established three categories of occult pneumothorax, based on size and location: (a) minuscule (< 1 cm in greatest thickness, seen on four or fewer images); (b) anterior (> 1 cm in greatest thickness, but not extending beyond the midcoronal line); (c) anterolateral (extending beyond the midcoronal line). In our study four of six patients with minuscule pneumothorax, including one who required mechanical ventilation, were observed without complications; two of six patients had chest tube placement. Seven of 14 cases with anterior pneumothorax were observed and resolved without complication; seven had chest tube placement. All eight patients with anterolateral pneumothoraces underwent percutaneous tube thoracostomy, regardless of proposed management. PMID- 8419441 TI - Thoracic Kaposi sarcoma in AIDS: CT findings. AB - Cases of acquired immunodeficiency syndrome (AIDS) related Kaposi sarcoma (KS) were reviewed to characterize the spectrum of thoracic findings seen with chest CT. Of 15 patients with AIDs-related KS involving the chest, 13 (87%) demonstrated pulmonary parenchymal disease characterized by multiple, bilateral flame-shaped or nodular lesions with ill-defined margins distributed along bronchovascular bundles. Pleural disease was noted in 10 (67%) patients, characterized by 9 cases of pleural effusions and 1 case of pleural implants. Chest wall disease involving the sternum, ribs, thoracic spine, and/or subcutaneous tissue was noted in eight (53%) patients. Although pleural and parenchymal lung disease are recognized manifestations of thoracic KS, there is also a high incidence of extrapulmonary chest disease evident on CT in patients with AIDS. PMID- 8419442 TI - Postoperative atrophy of posterolateral chest wall musculature: CT demonstration. AB - We review five cases in which CT demonstrated severe unilateral atrophy of the latissimus dorsi muscle. In four of these cases, similar findings were also seen in the inferior portion of the serratus anterior muscle. All patients had undergone thoracic surgery on the ipsilateral side 10 months to 3 1/2 years prior to CT. All patients had undergone posterolateral thoracotomy, including one who had undergone successive right and left posterolateral thoracotomies. The most likely explanation for the CT finding of atrophy is denervation injury resulting from surgical incision. We discuss normal innervation of the posterolateral chest wall musculature, with emphasis on the relationship of unilateral atrophy of posterolateral chest wall musculature to previous surgical incision. PMID- 8419443 TI - MRI of liver metastases from colorectal cancer vs. CT during arterial portography. AB - A prospective study was performed to compare, with a lesion-by-lesion analysis, the sensitivities of high field strength MRI and CT during arterial portography (CTAP) in detecting hepatic metastases from colorectal cancer. Twenty-one patients with liver metastases from colorectal cancer were prospectively investigated by high field strength MRI (1.5 or 2 T) and CTAP. High field strength MRI was performed with pre and post gadopentetate dimeglumine enhanced T1-weighted SE sequences and T2-weighted SE sequences. All patients underwent partial hepatectomy and 37 metastases were surgically and pathologically proved. The metastasis detection rate (sensitivity) was 94% (35 of 37) for CTAP and 78% (29 of 37) for high field strength MRI. The 16% (95% confidence interval: 1-31%) difference in sensitivity between CTAP and high field strength MRI was statistically significant (p < 0.05, McNemar test). The use of gadopentetate dimeglumine did not improve the sensitivity of T1-weighted SE sequences. Since our study demonstrated significant difference in sensitivities between high field strength MRI and CTAP in our group of patients, we can conclude that high field strength MRI cannot replace CTAP in the preoperative evaluation of patients with liver metastases from colorectal cancer. Computed tomography during arterial portography must be considered as the preoperative gold standard. PMID- 8419444 TI - Hydatid cyst of the liver: comparison of CT and MRI. AB - Twenty-seven cases of proven liver hydatid cysts were examined by MR and CT. On MR the cyst wall showed a rim of low signal intensity on both T2- and T1-weighted imaging. This capsule was more easily identified by MR than by CT. Specific signs of daughter hydatid cysts and detachment of the membrane were also evident by CT and MR examinations. Magnetic resonance was more effective than CT in detecting complications of the cyst; the first echo of T2-weighted imaging showed a very high signal of the hydatid content in cysts that communicated with the biliary tree or in superinfected cysts. However, MR did not help characterize solid or pseudotumoral forms of hydatid disease. PMID- 8419445 TI - MRI of adenomatous hyperplastic nodules of the liver in Budd-Chiari syndrome. AB - We present two cases of adenomatous hyperplastic nodules (AHN) occurring in patients with chronic Budd-Chiari syndrome who were investigated with MRI. In one case the foci of AHN were isointense to the liver on spin echo (SE) T1-weighted MRI and were hyperintense on both SE proton density and SE T2-weighted MRI. In the other case the nodules were hyper-intense on SE T1-weighted MRI. They were isointense on SE proton density-weighted MRI and became slightly hyperintense on SE T2-weighted MRI. Furthermore, in both cases one nodule displayed hyperintensity with hypointense rims on SE T2-weighted MRI. PMID- 8419446 TI - Splenic abscesses in typhoid fever: US and CT studies. AB - Salmonella typhi splenic abscesses (SA) are traditionally considered to be a rare complication of typhoid fever (TF). Our prospective study in an extensive series of patients was aimed at demonstrating that the incidence of SA was usually underestimated. Four hundred patients were systematically examined by abdominal ultrasound (US) at the time of proven biological diagnosis of TF, which was the 2nd week of admission. Twenty patients with persistent or recurrent symptoms (despite adequate treatment) or with acute lower thoracic pain were followed by US and CT. In eight patients both imaging modalities revealed anomalies compatible with SA. PMID- 8419447 TI - Dynamic CT in pancreatic lymphoma. AB - We retrospectively reviewed the dynamic CT examinations of eight patients with pancreatic lymphoma. Four tumors were rounded masses with well-defined contours, four were more infiltrating lesions. The median cross-sectional diameter of the tumors was 6 cm (range 2.5-12 cm). At dynamic CT, the tumors were hypodense (n = 8) and somewhat heterogeneous (n = 6). Additional features were enlarged lymph nodes, 1-3 cm in diameter (n = 5), dilatation of the biliary tract and pancreatic duct (n = 5), abnormalities in the fat around the celiac trunk and/or the superior mesenteric artery (n = 4), and venous stenosis or occlusion (n = 7). The CT findings of pancreatic lymphoma are more various than has been previously reported. Findings such as small tumor size, well-defined contours, tumor heterogeneity, pancreatic duct dilatation, and venous invasion may be seen. Pancreatic lymphoma cannot be reliably distinguished from pancreatic carcinoma by CT findings alone. PMID- 8419448 TI - CT pixel mapping in the diagnosis of small angiomyolipomas of the kidneys. AB - In six small renal angiomyolipomas (7-17 mm) the superiority of displaying the CT numbers of pixels within a lesion (pixel mapping) over the usual region of interest (ROI) measurement is described in the detection of small amounts of fat tissue. On precontrast 5 mm CT the ROI measurements were > 0 in four cases whereas pixel maps revealed pixels with values < 0 in six cases. PMID- 8419449 TI - IADR abstracts special issue. Chicago, Illinois, March 10-14, 1993. PMID- 8419450 TI - The AIDS epidemic. Challenges for nursing homes. AB - Caring for persons with AIDS and HIV-related illness is extremely rare within long-term care facilities. A survey of 54 Illinois nursing facilities indicated that homes were reluctant to care for persons with AIDS due to concerns about cost, staff capabilities or resistance, and concerns about mix with other residents. Although nursing homes are often blamed for the limited care provided for persons with HIV infection, this study indicated that the demand on facilities to provide this care is quite limited to date. PMID- 8419451 TI - Pressure ulcer risk assessment tools. What's new for gerontological nurses. AB - Impaired skin integrity is one of the most common and most costly problems facing the institutionalized elderly. Risk assessment tools provide the nurse with an efficient and systematic means of identifying those clients at high risk for pressure ulcer development. Pressure ulcer risk assessment tools are usually based on a combination of the following variables: mobility, continence, mental status, activity, nutrition, or physical status. Further testing and comparison of the various pressure ulcer risk assessment tools in the elderly population is needed to establish relative scientific and practical merit. PMID- 8419453 TI - Changes in CLIA limit screening tests. PMID- 8419452 TI - Is there a difference? Nursing in proprietary and nonprofit nursing homes. AB - Nurses working in proprietary and nonprofit nursing homes allocate their time in similar ways across various job responsibilities. Nurses who work in nursing homes obtain health histories and perform physical and psychosocial assessments on very few patients, regardless of home ownership. Compared with nurses who work in nonprofit nursing homes, nurses who work in proprietary homes perform significantly more physical examinations on their patients. Nurses employed by proprietary and nonprofit nursing homes receive similar hourly wages, but those employed in proprietary nursing homes receive fewer fringe benefits. PMID- 8419454 TI - Interpretation of abnormal laboratory values in older adults. PMID- 8419455 TI - Research considerations. Geropsychiatry unit evaluation. AB - The specialized psychiatric units for geriatric patients are believed to be effective, yet no studies could be found documenting this. A method of evaluating the effectiveness of a geropsychiatric program is to measure changes in the cognitive and functional status of its patients. This study supports the hypothesis that there would be an improvement in the geropsychiatric patients' cognitive and functional assessment scores by discharge and after hospitalization. PMID- 8419456 TI - Immunocytochemical evidence that S-100-positive cells of the mouse anterior pituitary contain interleukin-6 immunoreactivity. AB - We have previously shown that bioactive interleukin-6 (IL-6) is produced by rat and mouse (anterior) pituitary cells in vitro. Since the amount produced correlated with the presence of S-100-containing folliculostellate (FS) cells, these cells were suggested to be a source of IL-6 in the anterior pituitary (AP) lobe. In the present study we used immunocytochemical techniques to confirm this presumption. Freshly isolated mouse pituitary cells were subjected to immunocytochemical procedures whereby two different (neutralizing) monoclonal antibodies (MAb) against mouse IL-6 (6B4 and 20F3) and a polyclonal antiserum raised against bovine S-100 were used as primary antibodies. Single immunostaining revealed a small portion of mouse pituitary cells (about 6.5%) to be positive for IL-6 immunoreactivity with both antibodies. Importantly, the same proportion of cells was found to be IL-6 positive if only the AP was used as the cell source. About 7.5% of the pituitary cells stained for the presence of S-100 immunoreactivity. Positive staining for IL-6 was also found in pituitary cell samples from 2-day-old monolayer cultures and from redispersed 9-day-old histotypic aggregates, which both secreted bioassayable IL-6. In contrast, no IL 6 staining was found in AtT-20 cells, an established ACTH-secreting tumor cell line of the mouse pituitary which did not secrete bioactive IL-6. The specificity of the IL-6 immunostaining was demonstrated by a total loss of staining when MAb 6B4 was omitted or replaced by irrelevant rat IgG. Conclusively, pre-adsorption of the anti-IL-6 MAb (6B4) with recombinant mouse IL-6 totally abolished staining of pituitary cells. Double immunostaining for IL-6 and S-100 revealed that most if not all of the IL-6-containing pituitary cells were positive for S-100. Few of the S-100-containing cells did not stain for IL-6. These results confirm our previous hypothesis that FS cells, characterized by immunostaining of S-100 protein, contain bioactive and immunoreactive IL-6 and therefore are very likely producers of IL-6 in the AP. Furthermore, our results suggest that IL-6 is implicated in the local regulatory role ascribed to FS cells in the pituitary gland. PMID- 8419457 TI - Sensitive detection of rat gastrin mRNA by in situ hybridization with chemically biotinylated oligodeoxynucleotides: validation, quantitation, and double-staining studies. AB - Chemically biotin-labeled oligonucleotides form attractive reagents, as large quantities of stable and well-defined probes can easily be produced. Their usefulness for in situ hybridization was tested using rat gastrin cells as a model. Two probes recognizing two different regions of rat gastrin mRNA were synthesized and produced specific and equally strong hybridization signals. A probe complementary to human gastrin mRNA, but with mismatches to the rat gastrin mRNA sequence, failed to reveal rat gastrin cells under the stringency conditions used. Northern blotting revealed that the rat gastrin mRNA probes reacted exclusively with the appropriately sized (approximately 650 bases) mRNA. Model systems demonstrated that the hybridization signal, as revealed by alkaline phosphatase-based detection, varied linearly with the 10logarithm of target concentration and also showed that a new detection system was much more sensitive than previously used systems. In agreement with previous biochemical data, image analysis showed that starvation of rats led to a progressive decrease in cell staining intensities and cell numbers. Double staining for rat gastrin mRNA and gastrin immunoreactivity showed that in adult rats almost all gastrin cells expressed both mRNA and protein. Similar studies on developing rat gastrin cells revealed discrepancies between gastrin mRNA and gastrin-immunoreactive cells during the first week of newborn life. Subsequently, expression of mRNA and protein in the cells became gradually more concordant. PMID- 8419458 TI - Bone sialoprotein (BSP) secretion and osteoblast differentiation: relationship to bromodeoxyuridine incorporation, alkaline phosphatase, and matrix deposition. AB - We defined two distinct maturational compartments (proliferative and secretory) of osteogenic cells in vivo on the basis of ALP activity, BrdU incorporation, cell shape, and BSP production. BSP immunoreactivity was found to mark cells in the secretory but not in the proliferative compartment. We established the phenotypic similarity of primitive marrow stromal cells with proliferating perichondral cells (fibroblast-like, ALP+, BrdU+, BSP-). This suggests the potential functional equivalence of the two cell types as committed non-secretory osteogenic cells and points to the duality of osteogenic cell compartments as a generalized feature of bone formation. We further showed that although BSP secretion is a hallmark of the onset of osteogenesis, BSP antigenicity is lost both in osteoid and in a large proportion of mature osteoblasts during subsequent phases of bone deposition. This suggests that bone formation may not be a uniform event, as bone cells actually deposit antigenically, and likely biochemically, distinct matrices at specific times. PMID- 8419459 TI - Localization of bone sialoprotein (BSP) to Golgi and post-Golgi secretory structures in osteoblasts and to discrete sites in early bone matrix. AB - Bone sialoprotein (BSP), a bone matrix-enriched glycoprotein containing the Arg Gly-Asp (RGD) motif and endowed with cell binding properties, was localized in osteoblasts and early bone matrix of developing rat bone at the ultrastructural level. Preliminary light microscopic observations indicated that intracellular labelling was restricted to a paranuclear dot corresponding to the "negative Golgi image" of classical histology. The same pattern was observed whether antisera against the fully glycosylated protein or a peptide antiserum to a stretch of amino acids in human BSP sequence were employed. At the EM level, we obtained labeling over the Golgi area of osteoblasts but not over the rER. The labeling was concentrated over distensions of the trans Golgi and over pro secretory granules. In the matrix, BSP was distributed in a non-random manner. The label was concentrated over spherical aggregates of finely fibrillar material corresponding to the sites of early mineral deposition (so-called "mineralization nodules"). Such BSP-positive foci were seen both close to and away from the cell surface. The predominant association of BSP with Golgi and post-Golgi secretory structures and its absence from rER, as well as the reproducibility of the same pattern of localization with different antisera, might indicate a slow transit of the protein through the Golgi, not necessarily associated with protein glycosylation. PMID- 8419460 TI - Distribution of the slow/cardiac isoform of skeletal muscle Ca(2+)-ATPase in developing and mature tissues of chickens determined using a monoclonal antibody. AB - We have established that the monoclonal antibody (MAb) AA21, raised against a crude sarcolemmal fraction prepared from adult chicken anterior latissimus dorsi muscle, recognizes the slow twitch/cardiac isoform of calcium ATPase. This was done using a combination of immunohistochemistry at the light and electron microscopic level, the change in the cell distribution in skeletal muscle during development, the molecular weight of the principal protein recognized in Western transfers, and direct comparison with another MAb of known specificity. The antigen is initially expressed by all myotubes at E10 and with development is gradually lost from all presumptive fast fibers. In addition to its immunoreaction and slow extrafusal skeletal muscle fibers, AA21 displays a highly selective immunoreactivity with a number of other cell types in different tissues. The antibody stains a subset of intrafusal muscle fibers and intestinal and arterial smooth muscle, but not venous smooth muscle. In the nervous system, a subpopulation of neurons is intensely stained, most neurons are faintly stained, and glia are not stained at all. PMID- 8419461 TI - Production of a monoclonal antibody against cell-surface glycoprotein of guinea pig adrenocortical cells. AB - A monoclonal antibody (MAb) that reacted with the cell-surface antigens of adrenocortical cells was generated against cell suspensions from guinea pig adrenal glands. Cell-surface membranes of the adrenocortical cells in all zones, i.e., zona glomerulosa, zona fasciculata, and zona reticularis, were labeled with the antibody. Adrenal medulla remained unlabeled. Immunoelectron microscopy showed that entire plasma membranes, i.e., plasma membranes between adjacent cells and free cell-surface membranes, including sinusoidal microvilli, were immunoreactive to the antibody. Immunoblot analysis demonstrated that the antibody bound to two prominent bands at molecular weights of approximately 62,000 and 110,000. Two bands were stained with lectin-digoxigenin conjugates. The 110 KD band reacted with Datura stramonium (DSA) and Maackia amurensis (MAA) agglutinins, indicating the presence of N-acetyl-glucosamine and sialic acid linked alpha (2-3) to galactose; the 62 KD band reacted with SNA, indicating the presence of sialic acid-linked alpha (2-6) to galactose. In adrenocortical cells, the reaction pattern of Sambucus nigra (SNA) agglutinin was similar to that of the (MAb), whereas reaction patterns of DSA and MAA were different. Both neuraminidase digestion and prior absorption of the antibody with N-acetyl neuraminic acid completely prevented the immunolabeling of adrenocortical cells. These results indicate that the MAb mainly recognizes the 2-6 sialylated cell surface antigen of adrenocortical cells. PMID- 8419462 TI - Phagocytosis of outer segments by retinal pigment epithelium: phagosome-lysosome interaction. AB - We studied phagocytosis of rod outer segments (ROS) by the retinal pigmented epithelium (RPE) using rapid freezing, freeze-drying, and electron microscopic immunocytochemistry. Phagocytosis of photoreceptor outer segment tips by the RPE occurs daily, and in rats the shedding of these tips is light-entrained to a circadian rhythm. We studied the phagocytic process 5, 30, 90, and 150 min after light onset or after subjective light onset in rats entrained to a 12-hr dark-12 hr light cycle. Lysosomes were labeled with antibodies to cathepsin D, a major lysosomal enzyme responsible for opsin degradation. Phagosomes and phagolysosomes were recognized because of the lamellar structure of their photoreceptor-derived contents. We found a population of lysosomes that fuse with one another before they interact with phagosomes. This fusion can be triggered either by light or by endogenous circadian mechanisms. We also found that lysosome-phagosome interaction occurs after the ingestion stage is completed and that this interaction occurs in two steps. First, smaller lysosomes fuse with phagosomes. Subsequently, larger lysosomes appear to interact with phagosomes via pore-like or bridge-like structures. It is proposed that interchange of contents takes place through these structures. PMID- 8419463 TI - Cysteine proteinases in rat parathyroid cells with special reference to their correlation with parathyroid hormone (PTH) in storage granules. AB - To further understand the roles of storage granules in parathyroid cells, we examined by immunocytochemistry the localization of cathepsins B and H and of PTH in rat parathyroid gland. In semi-thin sections, small and large granular immunodeposits for cathepsins B and H appeared in the cells, whereas those for PTH were detected throughout the cells, especially in perinuclear regions. By electron microscopy, immunogold particles indicating cathepsins B and H labeled lysosomes and storage granules, whereas those showing PTH were localized in storage granules, small secretory granules, and the trans-Golgi network. Small vesicles labeled by immunogold particles showing these proteinases often appeared close to the storage granules. By double immunostaining, immunogold particles indicating these proteinases were co-localized with those for PTH in storage granules. By EDTA treatment, immunoreactivity for cathepsins B and H and for PTH was notably reduced in the cells, but immunoreactivity for the proteinases was still seen in lysosomes. These results suggest that storage granules in the rat parathyroid cells fuse with small vesicles containing cathepsins B and H, which may participate in regulating the intracellular PTH levels by degrading PTH in the granules. PMID- 8419464 TI - Hypo-osmotic shock induces an osmolality-dependent permeabilization and structural changes in the membrane of carp sperm. AB - We carried out spectrofluorimetric and flow cytometric measurements to investigate the effect of hypo-osmotic shock on cell membranes of common carp sperm. The time course of the permeability of the sperm cell membrane, as monitored by DNA-related propidium iodide (PI) fluorescence, was followed for 30 min after dilution of semen in hypo-osmotic environments of different ionic strengths. Spectrofluorimetric measurements indicated a continuous increase in the total PI emission intensity of a sperm suspension. Cell-by-cell flow cytometric measurements suggested that the permeability changes were of the all or-none type. The permeabilized fraction of cells in the individual samples was time and osmolality dependent. The number and percentage of cells in which DNA was stained by PI increased gradually over time and reached a steady-state plateau value after 5-15 min. This equilibrium fraction of cells with a PI permeable cytoplasmic membrane displayed an inverse relationship with the osmolality of the diluent, having a near 100% value for fresh water and distilled water. Dilution of sperm in hypo-osmotic medium brought about a fast decrease in the forward light-scattering signal on a short time scale compared to the pre steady-state time of the permeabilization. With the addition of extracellular Ca2+ (1.8 mM), restoration of the light scattering signal was observed. Permeabilization of the membrane and restoration of light scattering were not coincident in time. We propose a two-dimensional reorganization of the lipid structure as the underlying mechanism of the latter process. PMID- 8419465 TI - Immunohistochemical localization of carboxylesterase in the nasal mucosa of rats. AB - The enzymatic esterase activity of carboxylesterases is integral to the nasal toxicity of many esters used as industrial solvents or in polymer manufacture, including propylene glycol monomethyl ether acetate, dimethyl glutarate, dimethyl succinate, dimethyl adipate, and ethyl acrylate. Inhalation of these chemicals specifically damages the olfactory mucosa of rodents. We report the localization and differential distribution of a 59 KD carboxylesterase in nasal tissues of the rat by immunohistochemistry. Rabbit antiserum against the 59 KD rat liver microsomal carboxylesterase bound most prominently to the olfactory mucosa when applied to decalcified, paraffin-embedded sections of rat nasal turbinates. Within the olfactory mucosa, anti-carboxylesterase did not bind to sensory neurons, the target cell for ester-initiated toxicity; these cells apparently lack carboxylesterase. Instead, the antibody was preferentially bound by cells of Bowman's glands and sustentacular epithelial cells which are immediately adjacent to the olfactory nerve cells. In contrast, non-olfactory tissues (respiratory mucosa and squamous epithelium), which are more resistant to the toxicity of esters, had less carboxylesterase content. The distribution of immunoreactivity correlated well with the distribution of carboxylesterase catalytic activity described elsewhere. These findings help to link the metabolic fate of inhaled esters to the site-specific pathological findings that follow exposure to such chemicals. PMID- 8419466 TI - A novel fluorogenic substrate for detecting alkaline phosphatase activity in situ. AB - We describe here the in situ detection of alkaline phosphatase (APase) activity with a new fluorogenic substrate, 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4 (3H)-quinazolinone (CPPCQ). CPPCQ is very soluble and colorless. APase converts it into a rapidly precipitating product, whose strong fluorescence marks the sites of APase activity. The detected APase was either a probing enzyme anchored to epidermal growth factor (EGF) receptors of fixed human epidermoid carcinoma cell line (A431) by biotinylated EGF and streptavidin-APase conjugates or an endogenous marker existing in a fixed canine kidney cell line (MDCK). With CPPCQ staining, the EGF receptors and the endogenous APase were both visualized by fluorescence microscopy as contrasting, photostable, and well-resolved fluorescent stains. The EGF receptor staining was specific since it could be blocked by excessive unlabeled EGF. In contrast, fluorescein-labeled EGF failed to specifically stain the EGF receptors under the same fluorescent microscope. The endogenous APase staining with CPPCQ was sensitive to heating, levamisole and L-homoarginine, showing an APase tissue specificity of the liver/bone/kidney type. Therefore, CPPCQ appears to be a novel substrate dye for sensitive fluorescence APase histochemistry. PMID- 8419467 TI - Regulation of IFN-gamma and tumor necrosis factor-alpha expression in vivo. Effects of cycloheximide and cyclosporine in normal and lipopolysaccharide treated mice. AB - Despite accumulating information about cytokine expression in vitro, relatively little is known about the regulation and biologic relevance of these mediators in vivo. In order to study the effects of inhibition of protein synthesis and cyclosporine in vivo, we made use of systemically administered LPS, which induces the expression of a variety of cytokines. The expression of IFN-gamma and TNF alpha mRNA in normal and LPS-treated mice was examined by Northern blot analysis and amplification using the polymerase chain reaction. IFN-gamma activity was monitored using the biologic end point of MHC induction. TNF-alpha activity in serum was assessed using a L929 cytotoxicity assay. Messenger RNA for IFN-gamma and TNF-alpha could not be reliably detected by Northern analysis in spleens or kidneys of normal mice. After treatment with cycloheximide, a protein synthesis inhibitor, IFN-gamma and TNF-alpha mRNA could be detected in both sites in otherwise normal mice. The level of both IFN-gamma and TNF-alpha mRNA increased after LPS, although the temporal patterns of expression were different. The concurrent administration of cycloheximide led to marked superinduction of both cytokine mRNA levels. Similar effects were seen in T cell-deficient nude mice, suggesting that these responses are T cell independent. Cyclosporin A blocked induction of IFN-gamma in a dose dependent manner, but failed to significantly inhibit TNF-alpha mRNA or protein expression. Thus at least part of the immunosuppressive effect of cyclosporin A in vivo may be caused by its ability to inhibit the expression of certain cytokine genes, as has been found in vitro systems. However, the cellular target for this effect may extend to cell populations other than T cells. PMID- 8419468 TI - Human IL-10 is produced by both type 1 helper (Th1) and type 2 helper (Th2) T cell clones and inhibits their antigen-specific proliferation and cytokine production. AB - IL-10 gene transcription and IL-10 protein production was assessed in both type 1 (Th1) and type 2 (Th2) CD4+ human T cell clones by polymerase chain reaction and ELISA, respectively. Although Th2 clones apparently showed higher IL-10 mRNA levels, IL-10 mRNA expression was consistently found in Th1 clones, as well. Likewise, measurable IL-10 levels were found in the supernatants of both Th1 and Th2 clones. The effect of human IL-10 (h-IL-10) and viral IL-10 (v-IL-10) on the proliferative response and cytokine production by Th1 and Th2 human clones was also investigated. Addition in culture of h-IL-10 and v-IL-10 significantly reduced the proliferation of both Th1 and Th2 clones in response to the specific Ag and to PHA, but it had no inhibitory effect on the proliferative response of Th1 and Th2 clones to IL-2. h-IL-10 and v-IL-10 also inhibited the Ag-induced production of gamma-interferon (IFN-gamma) by Th1 clones and the production of IL 4 and IL-5 by Th2 clones, whereas they had no effect on the cytokine synthesis by the same clones stimulated with PMA plus anti-CD3 antibody. Preincubation of APC, but not of clonal T blasts, with h-IL-10 resulted in the inhibition of Ag-induced proliferation of both Th1 and Th2 clones, supporting the view that h-IL-10 primarily affects APC. These data demonstrate that, unlike the murine system where IL-10 is a product of Th2 (but not Th1) cells and seems to mainly down regulate the Th1 response, in the human system, IL-10 is produced by, and down regulates the function of, both Th1 and Th2 cells. PMID- 8419469 TI - Preferential activation of Th2 cells in chronic graft-versus-host reaction. AB - The injection of DBA/2 parental lymphocytes into adult, immunologically intact (C57BL/6 x DBA/2) F1 hybrid mice results in a chronic graft-vs-host reaction (GVHR) characterized by a deficiency in CD4+ T cell functions and a B cell activation leading to autoantibody production. The discovery that distinct subpopulations of Th cells may regulate the effector immune functions led us to investigate whether the chronic GVHR differentially affects Th subsets. Data are presented indicating that mice undergoing a GVHR spontaneously produced lymphokines of Th2 origin. IL-4 and IL-10 mRNA were detected in the spleens of GVH mice, and IL-4 was shown to be responsible for the increased expression of class II Ag on B cells. Moreover, upon polyclonal activation in vitro, GVH T cells exhibited defective IL-2 and IFN-gamma production but elevated IL-4 production. We conclude that the chronic GVHR is characterized by a selective deficiency in cells secreting IL-2 and IFN-gamma and a hyperactivation of Th2 cells. The simultaneous production of IL-4 and IL-10 might explain the association between B cell hyperactivity and impairment of Th1-like activities in various models that associate autoimmunity and immunosuppression, such as GVHR and HIV infection. PMID- 8419470 TI - Anergized T cell clones retain their cytolytic ability. AB - CD4+ T cells have been described to have both helper and lytic function. The helper function of Th1 cells in particular can be inactivated by inducing the T cell into a state of nonresponsiveness in which the T cell is no longer capable of producing IL-2 or proliferating in an autocrine way to a conventional antigenic stimulus. To determine whether the lytic ability of Th1 cells can also be rendered nonfunctional upon anergy induction, we induced Th1 clones into a nonresponsive state and tested their ability to lyse target cells in an Ag specific and MHC class II-restricted manner. We show that cells newly induced into an anergic state were able to lyse target cells nonspecifically. This effect was short-lived and after resting in culture media, the cells regained their ability to lyse target cells in an Ag/MHC-specific manner, and this ability was comparable to normal resting T cells. In contrast, the helper function of these cells remained nonresponsive, and the cells were unable to proliferate or to secrete IL-2 in response to the same antigenic stimulus used for lysis. Therefore, the lytic pathway appears to be regulated separately from the proliferative/lymphokine pathway(s) and is not affected long-term by an anergic stimulus. PMID- 8419471 TI - Targets of B lymphocyte antigen receptor signal transduction include the p21ras GTPase-activating protein (GAP) and two GAP-associated proteins. AB - Cross-linking membrane Ig (mIg) on B cells stimulates tyrosine phosphorylation of proteins involved in signal transduction including the mIg-associated proteins Ig alpha and Ig-beta, the tyrosine kinases p53/p56lyn, p55blk, p59fyn, and PTK72, phosphatidylinositol 3-kinase, phospholipase C gamma 1 and gamma 2, and the mitogen-activated protein kinase. We now show that the p21ras GTPase-activating protein (GAP) is also a substrate for mIg-activated tyrosine kinases. p21ras is a key regulator of cell growth and GAP may act as both a regulator of p21ras activity and as a downstream effector of p21ras. We found that mIg cross-linking caused a rapid increase in tyrosine phosphorylation of GAP in the immature B cell line WEHI-231, the mature B cell lines BAL 17 and Daudi, and the IgG-bearing B cell line A20. In fibroblasts, tyrosine kinase activation causes GAP to associate with two other tyrosine-phosphorylated proteins, p62 and p190, which have homologies to an RNA-binding protein and a transcriptional repressor, respectively. Similarly, mlg cross-linking induced the association of GAP with a 62-kDa tyrosine-phosphorylated protein in BAL 17, WEHI-231, and Daudi cells. Anti Ig treatment also increased the amount of a 190-kDa tyrosine-phosphorylated protein associated with GAP in WEHI-231 and Daudi cells. After separation by SDS PAGE and transfer to nitrocellulose, the tyrosine-phosphorylated p62 and p190 present in anti-GAP immunoprecipitates from B cells were capable of binding radiolabeled recombinant GAP, as previously reported for the GAP-associated p62 and p190 from fibroblasts. The amount of p62 that could be detected in this way after immunoprecipitation with antiphosphotyrosine antibodies was much greater from anti-IgM-treated BAL 17 cells than from unstimulated BAL 17 cells. This probably reflects anti-Ig-induced tyrosine phosphorylation of p62. In any case, GAP, p62, and/or p190 may be involved in signal transduction by mIg in B cells. PMID- 8419472 TI - IL-2-independent activity of IL-7 in the generation of secondary antigen-specific cytotoxic T cell responses in vitro. AB - Purified CD8+ splenocytes from influenza virus strain A/WSN/33 (H1N1)-immune BALB/c (H-2d) mice responded to a synthetic peptide, synthetic influenza nucleoprotein peptide 147-158 R-, with a sequence (147-158 R156-) derived from influenza A virus nucleoprotein with high affinity for Kd class I MHC molecules, with the generation of effector cytotoxic T (Tc) cells specific for influenza A virus-infected target cells in vitro. The process of the conversion of synthetic influenza nucleoprotein peptide 147-158R- -Kd-responding memory Tc into effector Tc cells requires Ag in the form of peptide associated with Kd class I MHC molecules and the presence of endogenously produced IL-2 by CD8+ T cells. Under blockade of utilization of endogenous IL-2 by mAb to IL-2 or IL-2R, no effector Tc cells were generated, but the addition of IL-7 restored the process of the conversion of CD8+ memory into effector Tc cells and significantly enhanced the specific cytolytic activity of Tc cells above those of controls. IL-7-reactive cells were found in both IL-2Rhigh- and IL-2Rlow-expressing responder CD8+ T cells. We conclude that the requirement for endogenous IL-2 in Ag-induced conversion of CD8+ memory Tc cells into effector Tc cells can be replaced by exogenous IL-7. This demonstrates that IL-7 is a potent regulatory cytokine with similar activity to IL-2 and may act independently of IL-2 in the Ag-specific activation of memory CD8+ Tc cells. PMID- 8419473 TI - Enhanced IL-4-mediated D10.G4.1 proliferation with suboptimal concentrations of anti-IL-4 receptor monoclonal antibodies. AB - Suboptimal concentrations of the anti-IL-4 receptor mAb M1 or M2 gave, in combination with IL-4, an enhanced proliferative response to the IL-4-responsive cell line D10.G4.1, compared to IL-4 alone. Increasing amounts of the M1 antibody inhibited the IL-4-dependent proliferation in a normal fashion. The enhanced IL-4 induced proliferation was only inhibited with the anti-IL-4 mAb 11B11, whereas the anti-IL-2 receptor antibodies had no effect. Addition of M1 or M2 antibodies to the IL-2-dependent cell line CTLL, known to express small amounts of IL-4 receptors and thereby also slightly responsive to IL-4, gave no enhanced IL-4 induced proliferation. Instead these antibodies were found to inhibit both the IL 4 as well as the IL-2-mediated CTLL proliferation. PMID- 8419474 TI - IL-4 induction of IgE class switching by lipopolysaccharide-activated murine B cells occurs predominantly through sequential switching. AB - Resting murine B cells activated with bacterial LPS co-express membrane (m)IgG1 and mIgE upon stimulation with IL-4. In this report, we combine both cellular and molecular approaches to elucidate the mechanism underlying this co-expression. We demonstrate that an anti-IgG1 antibody specifically and selectively inhibits IgE secretion (approximately 70%) by LPS + IL-4-stimulated B cells, which provides functional evidence for mIgG1 expression by precursors of IgE-secreting cells. The IgG1 and IgE secretory responses are separated temporally by approximately 16 h, with IgE production developing later than IgG1. A similar delay is observed in the appearance of mIgE+ cells suggesting that class switching to IgG1 precedes that to IgE. In the sort-purified, mIgG1+mIgE+ B cell population approximately 25% of cells expressed cytoplasmic (c) (secretory) IgG1 and approximately 15% expressed cIgE at the time of their isolation. However, only a small percent of the mIgG1+mIgE+ cells co-expressed cIgG1 and cIgE, further suggesting a temporal separation in IgG1 and IgE secretion within individual cells, but indicating that single cells can co-secrete these two Ig isotypes. Furthermore, the absolute level and rate of increase of IgG1 secretion by mIgG1+mIgE+ cells, upon their isolation and reculture, is lower than that for mIgG1+mIgE- cells suggesting a loss of CH gamma 1 expression in the former population. Analysis of total, unselected circular DNA excision products in LPS + IL-4-activated B cells demonstrates that most, if not all, of the DNA encoding the IgG1 constant heavy gene (CH gamma 1) (i.e., products of a class switch to IgE) have been rearranged. Collectively this data provides strong evidence at both the cellular and molecular level that the predominant mode of switching to IgE in response to in vitro stimulation by LPS + IL-4 is from IgM to IgG1 to IgE. PMID- 8419475 TI - Identification and propagation of a putative immunosuppressive orphan parvovirus in cloned T cells. AB - A putative parvovirus related to minute virus of mice (MVM), but distinct from MVM-prototype and MVM-immunosuppressive, was identified, using serologic techniques and Southern blot analysis, in maintenance cultures of established T cell clones. This putative viral agent resulted in a lytic infection of cloned L3 cytotoxic T cells but was unable to produce a productive infection in BHK.21 or EL-4(G) cells. Moreover, maintenance cultures of several distinct subsets of cloned T cells apparently contaminated with this putative viral agent contained poorly growing cells and erythrocyte aggregates. The aggregation of mouse erythrocytes appeared to be a reliable indicator of infection with this putative virus and may be related to the ability of this agent to agglutinate mouse erythrocytes. This putative virus also was found to inhibit the proliferative response of certain cloned T cells to IL-2 and Ag. Viremic mice and secondary MLC supernatant were identified as two potential sources of contamination and represent ways of propagating this agent in vitro. The finding that this agent interferes with the ability of T cell clones to thrive and, therefore has the potential to alter immune responses, emphasizes the importance of identifying and excluding parvoviral infections in cultures of murine T lymphocytes. PMID- 8419476 TI - Characterization of antigen-presenting cells that present exogenous antigens in association with class I MHC molecules. AB - Exogenous Ag in the extracellular fluids do not gain access to the class I Ag presenting pathway in most cells. However, there is an APC resident in spleen that can process and present exogenous Ag in association with class I molecules. We characterize the phenotype of this cell. This APC is of low buoyant density, is adherent to Sepharose and glass, and expresses both class II molecules and FcR. This phenotype identifies this APC as a macrophage. Resident, peptone- and thioglycolate-induced peritoneal macrophages also display this Ag-presenting activity. Analysis with CTL clones suggest that this Ag-presenting pathway may be active in only a subset of macrophages. A similar Ag-presenting activity is also present in dendritic cell-enriched populations from spleen although we cannot rule out the possible involvement of contaminating macrophages. In contrast, B and T cells that are resident in spleen and LPS blasts are unable to present exogenous Ag in association with class I molecules. The presentation of exogenous OVA with class I molecules is not inhibited by the inhibitors of thiol proteases, leupeptin, and antipain. The presence of gelonin, a ribosomal inactivating protein, in the extracellular fluids inhibits the ability of these APC to present exogenous OVA. Under identical conditions, gelonin does not inhibit Con A stimulated T cell proliferation, or LPS-stimulated B cell proliferation and Ag presentation. These results are discussed in relation to the potential pathways through which an Ag in the extracellular fluids is presented with MHC class I molecules. PMID- 8419477 TI - Origin, differentiation, and repertoire selection of CD3+CD4-CD8- thymocytes bearing either alpha beta or gamma delta T cell receptors. AB - It has been widely accepted that CD3+CD4-CD8- T cells expressing TCR-alpha beta or TCR-gamma delta (found in the thymus as well as in the periphery) represent lineages distinct from either CD4+CD8- and CD4-CD8+ single-positive T cells expressing TCR-alpha beta. However, the origin, differentiation pathway, and TCR repertoire selection of CD3+CD4-CD8- T cells remain controversial. We demonstrate that CD3+CD4-CD8- thymocytes can be separated into three subsets based on their expression of heat-stable Ag (HSA) and CD44. Our results further suggest the following: 1) the HSA+ subset represents a pre-selection population, although the HSA- subset is a postselection subset; 2) the high incidence of V beta 8.2 usage among CD3+CD4-CD8- thymocytes is a result of positive selection, rather than a predetermined event at a precursor cell level; 3) the maturation of CD3+CD4-CD8- thymocytes proceeds along the following differentiation pathway: HSA+CD44(-)- >HSA-CD44(-)-->HSA-CD44+. Both TCR-alpha beta +CD4-CD8- and TCR-gamma delta +CD4 CD8- thymocytes show similar differentiation processes; 4) CD3+CD4-CD8-cells directly differentiate from CD25+CD3-CD4-CD8- thymocytes which include precursor cells for both the CD3+CD4-CD8- and the CD4+CD8-/CD4-CD8+ lineages. Taken together, these results suggest that the CD25+CD3-CD4-CD8- stage of thymocyte differentiation represents a branching point for either the CD4+CD8- or CD4-CD8+ single-positive lineages or the CD3+CD4-CD8- lineages. PMID- 8419478 TI - Molecular cloning and characterization of human eosinophil Charcot-Leyden crystal protein (lysophospholipase). Similarities to IgE binding proteins and the S-type animal lectin superfamily. AB - We have isolated and sequenced a 598-bp full length cDNA clone for the human Charcot-Leyden crystal (CLC) protein (eosinophil lysophospholipase), the unique and prominent constituent of human eosinophils and basophils that forms the hexagonal bipyramidal crystals classically observed in tissues and secretions from sites of eosinophil-associated inflammation. A 426-bp open reading frame encoded a 142-amino acid polypeptide with a predicted molecular mass of 16.5 kDa and isoelectric point of 7.28. The deduced amino acid sequence of CLC protein showed 20 to 30% similarity over regions of approximately 100 amino acids with the carboxyl-terminal domains of four IgE-binding proteins, including the 31-kDa human and rat IgE-binding proteins, the 35-kDa mouse carbohydrate binding protein (CBP35), Mac-2, the murine macrophage cell surface protein that is identical to CBP35, and the human homologue of Mac-2. These proteins are members of a superfamily of beta-galactoside binding S-type animal lectins, which includes a group of highly conserved 14-kDa lectins isolated from human lung, heart, placenta, bovine heart, chicken skin, mouse fibroblasts, and the electric organ of the electric eel; CLC protein also showed sequence similarities to these 14 kDa animal lectins, including conservation of 7 of 16 invariant amino acid residues thought to comprise the carbohydrate-binding domain of these proteins, with conservative amino acid changes at others; thus, CLC protein could potentially possess carbohydrate or IgE-binding activities. Northern analyses revealed an approximately 900-bp mRNA species that was present in peripheral blood eosinophils from patients with eosinophilia, basophils from patients with chronic myelogenous leukemia, and in HL-60 cells induced towards eosinophilic differentiation with B cell growth factor-II (IL-5) or granulocytic differentiation with DMSO, but was absent in neutrophils, monocytes, T cells, B cells, or HL-60 cells induced towards monocytic differentiation with vitamin D3. Southern analyses revealed a gene of approximately 5 to 6 kb in length. The cDNA clone and complete amino acid sequence data for CLC protein will facilitate structure-function analyses of its unusual hydrophobic properties, unique propensity for crystallization, lysophospholipase, and potential lectin-like activities. PMID- 8419479 TI - Role of mouse VH10 and VL gene segments in the specific binding of antibody to Z DNA, analyzed with recombinant single chain Fv molecules. AB - A plasmid vector was constructed for the expression of a single chain Fv domain of mouse mAb to Z-DNA (antibody Z22), which is encoded by VH10 and V kappa 10 gene family members along with Dsp2, JH4, and J kappa 4 segments. The vector coded for a PhoA secretion signal, VH segment, flexible peptide linker, VL segment, (His)5, and a protein A domain. Unique restriction sites allowed exchange of the segments as cassettes. Bacteria transformed with the vector secreted soluble recombinant Fv with specific Z-DNA-binding activity. When the L chain of Z22 was replaced with a library of splenic VL cDNA from a mouse immunized with Z-DNA, only a light chain closely resembling that of the original Z22 (differing at six amino acid positions) yielded Fv with Z-DNA-binding activity. The Fv with this L chain replacement had a lowered affinity, but remained selective for Z-DNA. Replacement of the Z22 H chain with a mixture of 11 VH10-encoded H chains yielded two Z-DNA binding clones, but they bound B-DNA and denatured DNA as well as Z-DNA. The replacement clones indicate the importance of the H chain CDR3 and particular VH-VL combinations in formation of specific antibodies to Z-DNA. PMID- 8419480 TI - Structural analysis of the CD11b gene and phylogenetic analysis of the alpha integrin gene family demonstrate remarkable conservation of genomic organization and suggest early diversification during evolution. AB - CD11b is a member of the beta 2 subfamily of the human leukocyte integrins. Its expression is limited to mature myeloid and NK cells and is up-regulated during the course of granulocytic and monocytic differentiation. The CD11b/CD18 (Mo1) heterodimer promotes adhesion of granulocytes and monocytes to C3bi-coated bacteria and endothelial cells. In an attempt to relate the exon structure to the known functional domains, as well as to identify and study cis-acting elements that are involved in its tissue-specific expression, we have isolated genomic clones encoding CD11b, deduced the exon/intron organization, and determined the transcriptional start site. The CD11b gene spans 55 kb and is encoded by 30 exons. Its structure closely resembles that of CD11c, another of the three leukocyte integrin alpha-chains, and suggests that these two genes arose by a gene duplication event. Furthermore, comparison of the CD11b gene structure with that of platelet glycoprotein IIb and Drosophila PS2 suggest how the human leukocyte integrins evolved and dispersed during the course of evolution. PMID- 8419481 TI - The human Ig-beta cDNA sequence, a homologue of murine B29, is identical in B cell and plasma cell lines producing all the human Ig isotypes. AB - The B cell Ag receptor complex consists of at least two disulfide-linked, heterodimeric structures: the clonally restricted membrane Ig (mIg) molecule and the nonpolymorphic Ig-alpha:Ig-beta protein dimer. The latter molecule is encoded by two separate genes, mb-1 and B29. The DNA sequences of murine and human mb-1 and murine B29 have been determined previously. This study describes the sequence of the full-length human cDNA homologue of the murine Ig-beta/B29 message. The human sequence codes for a protein that displays the typical subunit features of a transmembrane member of the Ig superfamily. The transmembrane and intracytoplasmic domains exhibit striking nucleotide and amino acid sequence similarity between the two species. These regions show almost complete conservation of areas presumed to be involved in noncovalent interactions with other members of the receptor complex and with intracellular kinases and cytoskeletal components. The only sequence dissimilarity seen in these presumed critical areas involves the Y-E-G-L-N motif, a potential target for tyrosine phosphorylation. In contrast, the extracellular portion is much more divergent. Inasmuch as similar patterns of species diversity have been reported for Ig alpha, the Ig-alpha and Ig-beta molecules may have coevolved to maintain species specific extracellular interactions between one another and with mIg. Similar to the Ig-alpha molecule, the Ig-beta sequence is identical in B lineage cells expressing all five Ig isotypes. However, in contrast to the Ig-alpha molecule, the Ig-beta sequence is expressed at apparently similar levels in terminally differentiated, mIg- plasma cells as well as in mIg+, mature B cells. These data suggest that Ig-beta has functions in addition to those associated with surface mIg expression. PMID- 8419482 TI - Characterization of endogenous peptides bound to purified HLA-DR molecules and their absence from invariant chain-associated alpha beta dimers. AB - We have examined peptides bound in vivo to DR alpha beta dimers and alpha beta Invariant (I) chain complexes purified directly from a DR11, DRw52 homozygous B lymphoblastoid cell line. Nine major peptides were purified by reversed-phase HPLC from cell derived alpha beta dimers and sequenced. Eight of these were from known sources, including both endogenously synthesized and exogenous proteins. The endogenously derived peptides originated from secretory proteins, from the extracytoplasmic regions of transmembrane proteins, and from heat shock proteins. All of the peptides were from 13 to 16 amino acids in length. Comparison of the sequences of a subset of these DR-associated peptides with those of other reported DR-binding peptides suggests that an aromatic amino acid followed by a basic amino acid seven residues C-terminal from it may provide a generalizable, but not absolute, DR-binding motif, with additional residues possibly contributing to DR allelic specificity. In contrast to the alpha beta dimers, no peptides were detected bound to purified alpha beta I complexes. These data suggest that I chain-associated alpha beta dimers within the cell do not bind peptides, and provide in vivo evidence that I chain prevents the binding of inappropriate peptides to class II molecules during early stages of transport. PMID- 8419483 TI - The analysis of internal image-bearing anti-idiotypic monoclonal antibody in relation to carcinoembryonic antigen. AB - Five anti-Id mAb (Ab2) were prepared from a BALB/c mouse immunized with anti carcinoembryonic Ag (CEA) mAb MA208 (Ab1) in a syngeneic system. These anti-Id mAb appear to recognize unique idiotopes at the combining site of mAb MA208, because they were specifically reactive with mAb MA208 and showed the inhibitory activity against the binding of mAb MA208 to CEA. These anti-Id mAb were divided into three groups: group 1 (M7-625), group 2 (M7-413, M7-914), and group 3 (M7 049, M7-418), according to the analysis of anti-anti-Id antibodies (Ab3) induced with each anti-Id mAb (Ab2). Anti-anti-Id mAb M7-625 antisera (Ab3) reacted with purified CEA in binding assay and in Western blot analysis, and competed with Ab1 binding to CEA. Furthermore, the binding of anti-Id mAb M7-625 (Ab2) to mAb MA208 (Ab1) was inhibited with CEA, indicating that Ab2 mimicks the structure of the epitope in CEA which was recognized with Ab1. These serologic findings suggest that anti-Id mAb M7-625 carries the internal image of the Ag. According to the amino acid sequences of CDR 1, 2, and 3 of the mAb M7-625 variable region, there exists a homology of amino acid sequences between CDR2 in the H chain (5 amino acids of 10) and CDR3 in the L chain (3 amino acids of 9) of mAb M7-625 and domain III of CEA (545-554). PMID- 8419484 TI - Bone marrow macrophages process exogenous Toxoplasma gondii polypeptides for recognition by parasite-specific cytolytic T lymphocytes. AB - CD8+ T cells from mice vaccinated with an attenuated strain of Toxoplasma gondii have previously been shown to have cytolytic activity against bone marrow macrophages (BMM phi) preincubated with a soluble tachyzoite extract. In the present study, we show that class I-transfected L cells differ from BMM phi in that although both cell types are recognized CTL after infection, only BMM phi are killed after sensitization with soluble tachyzoite extract. Gel filtration studies indicated that the T. gondii Ag responsible for sensitization of BMM phi are macromolecules of M(r) > or = 12,000. In contrast, peptides derived by tryptic digestion of this material were found to sensitize both transfected L cells and BMM phi. Although exogenous beta 2-microglobulin markedly enhanced peptide sensitization of BMM phi, no such effect was observed using the macromolecular preparation. This result suggests a requirement for cellular internalization in the processing by BMM phi of soluble Ag for class I-restricted recognition. In related experiments, infected and Ag-sensitized BMM phi were found to express cross-reactive T. gondii epitopes, as determined by cold target inhibition studies. Supernatant derived by 100,000 x g centrifugation of tachyzoite extract had potent sensitizing activity, and after anion exchange chromatography most of the activity was associated with a single fraction. The p30 Ag was not detected by immunoblot analysis in the biologically active supernatant and chromatographic fractions. These findings establish the feasibility of identifying the parasite Ag recognized by CD8+ effectors by direct fractionation of T. gondii proteins coupled with sensitization of BMM phi targets. PMID- 8419485 TI - IgA antibodies to a protective antigen in human Schistosomiasis mansoni. AB - The specific IgA antibody responses to the protective recombinant Schistosoma mansoni 28-kDa glutathione-S-transferase (Sm28GST) Ag and to derived synthetic peptides have been evaluated before and 6 mo after chemotherapy in S. mansoni infected patients from Kenya. These studies revealed a parallelism between the age-dependent evolution of IgA antibody levels to Sm28GST and to one synthetic peptide (115-131) and the acquisition of resistance to reinfection. Functional analysis revealed that IgA antibodies to Sm28GST displayed a potent neutralizing effect on the enzymatic properties of the molecule, and also markedly impaired schistosome fecundity, by limiting both the egg laying of mature worms and the hatching capacity of schistosome eggs into viable miracidia. These results suggest that, in addition to IgE, IgA antibodies might participate in the protective immune response against schistosomiasis. PMID- 8419486 TI - IL-2 contributes to the IL-5 response in granulomas from mice infected with Schistosoma mansoni. AB - Th cells within the granulomas of murine schistosomiasis mansoni produce IL-5, which is essential for granuloma eosinophil growth and development. The mechanisms regulating granuloma IL-5 production are unknown. The granulomas also make IL-2 in small quantities. rIL-2 therapy stimulates eosinophilia and IL-5 synthesis. Therefore, we studied the effect of IL-2 on IL-5 production within the liver granulomas of murine Schistosoma mansoni. Dispersed granuloma cells and intact granulomas cultured in vitro released IL-5. Adding anti-IL-2 or anti-IL-2R to the cultures, to block IL-2 activity, significantly inhibited IL-5 production. However, supplementing the cultures with small quantities of rIL-2 markedly stimulated IL-5 release in a dose-dependent fashion. Blocking anti-IL-4 mAb had no effect. Also, granuloma T cells were isolated by FACS. These highly purified T cells produced IL-5 both in the presence and absence of plate-bound anti-CD3. Once again, the IL-5 production was dependent on IL-2. The requirement of IL-2 for normal IL-5 production was not dependent on an IL-2-induced expansion of the IL-5-producing, T lymphocyte population. Thus, IL-2 mediates T cell interactions within the granuloma that regulate granuloma IL-5 secretion. PMID- 8419487 TI - Repertoire diversity of antibody response to bacterial antigens in aged mice. III. Phosphorylcholine antibody from young and aged mice differ in structure and protective activity against infection with Streptococcus pneumoniae. AB - Aging in mice is accompanied by qualitative changes in the antibody repertoire to phosphorylcholine (PC), a natural epitope of certain pneumococci. The PC-specific mAb from young/adult (2-4 mo) BALB/c mice are uniformly encoded by the canonical IgV genes of T15 family, whereas the antibody from aged mice (> or = 20 mo) are molecularly heterogeneous, being encoded by various VH and VL genes of non-T15 families. Interestingly the young/adult and aged BALB/c mice produce comparable amounts of antibodies to PC regardless of the molecular shift in the antibody repertoire. This finding prompted our study on the relative ability of PC antibodies from young and aged donors to protect mice against virulent infection with type 3 pneumococci. Passive administration (i.p.) of pooled, PC-specific mAb generated from young donors (all from the T15 Ig gene family) protected the recipients against subsequent i.p. challenge with a lethal dose of Streptococcus pneumoniae strain WU2, in a dose-dependent manner. In contrast, a mixture of PC mAb from aged donors (all from non-T15 families) failed to protect the mice against the infection, even at the highest amount of administered (100 micrograms of mAb/recipient). Average affinity of the aged mAb for the free hapten (PC chloride) as well as their binding to the bacteria was lower than that of the young mAb. Similarly, affinity-purified serum PC antibody from S. pneumoniae vaccine-immunized young mice afforded a measurable degree of passive protection against the pneumococcal infection whereas a similar dose of serum PC antibody from aged mice did not. Further experiments showed that PC mAb from young donors were equally protective in either young or aged recipients challenged with the bacteria. These results demonstrate that the aged immune system may, in some instances, produce high levels of antibody that are structurally different and less protective against microbial infection. PMID- 8419488 TI - A role for gamma delta + T cells during experimental infection of mice with Leishmania major. AB - To assess the importance of gamma delta + T lymphocyte responses in experimental murine cutaneous leishmaniasis, the expression of gamma delta + TCR on the surface of lymphocytes in spleen and draining lymph nodes of mice infected with Leishmania major was examined. In both susceptible BALB/c and resistant CBA/J mice, an increase in gamma delta + T cells was observed after s.c. infection. Whereas in chronically infected BALB/c mice, these cells can represent up to 35% of the CD3+ cells in the spleen, in CBA/J mice the percentage of gamma delta + T cells returns to a lower level after resolution of lesions. Further experiments, in which the disease outcome was modified in susceptible BALB/c and in resistant CBA/J mice by chemotherapy or immunointervention, have confirmed that a correlation existed between the multiplication of the parasites and the expansion of gamma delta + T cells in the spleen. Interestingly, in BALB/c mice infected for 4 mo, these gamma delta + T cells represent 80% of the activated blast population and a large proportion of them express the alpha-chain of the CD8 molecule. In an attempt to assess a potential role for the gamma delta + T cells in the control of infection with L. major, the effect of injection of mAb against gamma delta TCR on the course of infection was studied. In both BALB/c and CBA/J mice, blocking of the gamma delta TCR resulted in the development of larger lesions that contained increased numbers of parasites. This treatment significantly delayed the healing of cutaneous lesions in otherwise resistant CBA/J mice. Taken together these results indicate that gamma delta + T cells are expanded during experimental infection of mice with L. major and could be involved in host defense against this parasite. PMID- 8419489 TI - Mobilization of different arachidonate pools and their roles in the generation of leukotrienes and free arachidonic acid during immunologic activation of mast cells. AB - Immunologic activation of mast cells leads to the mobilization of arachidonic acid (AA) from membrane phospholipids and the subsequent conversion of this AA to bioactive products. The objective of our study was to determine if segregated pools of AA-containing phospholipids within mast cells serve as independent sources of AA. Initial studies indicated that the appearance of free AA occurred rapidly (maximal level formed within 1 min) within supernatant fluids of Ag stimulated mast cells and was kinetically different from the formation of leukotriene (LT) B4 or LTC4. To examine whether free AA and leukotrienes were mobilized from different sources, AA-containing phospholipids of mast cells were labeled with [14C] and [3H] AA such that all major subclasses (1-acyl-1-alkyl-1 alk-1'-enyl) of phospholipids contained different ratios of [3H] to [14C] (sp. act. ratios (SAR)). Mast cells were then stimulated with Ag and the SAR of cellular AA, extracellular AA and extracellular LTC4, LTB4, 6-trans LTB4, were determined. The SAR were uniform in all LT and mimicked the ratio found in cellular AA. By contrast, the SAR of AA released into supernatant fluids was twofold lower than that of LT. This indicated that AA released as free fatty acid clearly was derived from a different lipid pool than AA that formed LT. Although it was apparent that the pools which gave rise to AA and LT were different, defining phospholipid(s) that constitute these distinct pools proved more difficult. The SAR of LT suggested that their cellular precursor AA could have been derived from several phospholipid subclasses; however, the SAR in phosphatidylcholine and phosphatidylinositol most closely matched the LT. The SAR of AA in supernatant fluids implied that it was derived, in part, from phosphatidylethanolamine subclasses. Taken together, these data suggest that there are at least two different pools of AA that are mobilized in mast cells during Ag activation. PMID- 8419490 TI - Activation of the C4 and C2 components of complement by a proteinase in serum bactericidal factor, Ra reactive factor. AB - Ra-reactive factor (RaRF) is a C-dependent bactericidal factor that binds specifically to LPS of Ra chemotype strains of Salmonella and kills the bacteria by triggering the C cascade. In the present study, we investigated the components of mouse RaRF that activate C4 and C2. The RaRF bound to LPS-coated E, and activated the C4 on the surface of E, causing the C4 to bind to the cells. Diisopropyl fluorophosphate (DFP) bound to RaRF and inhibited its ability to activate C4 and C2. Cleavage of the alpha-chain of C4 by RaRF generated a polypeptide with a size similar to that of the alpha'-chain of C4b, which is known to be a product of the cleavage of C4 by C1s subcomponent of C1. A fraction with the ability to activate C4 and C2 was separated from RaRF by gel-permeation chromatography in the presence of EDTA and acetonitrile. This fraction contained a DFP-binding polypeptide with an apparent m.w. of 100,000. This polypeptide is not the C1s in mouse C1 because the sizes of this polypeptide and of the fragments produced by its reduction were different from those of DFP-binding proteinases in mouse C1. These results indicate that mouse RaRF contains a C1s like serine proteinase that is capable of activating C4 and, probably, C2. PMID- 8419491 TI - Macrophage colony-stimulating factor in human fetal astrocytes and microglia. Differential regulation by cytokines and lipopolysaccharide, and modulation of class II MHC on microglia. AB - CSF-1 is a growth factor that selectively promotes the proliferation, survival, and differentiation of cells of the mononuclear phagocyte series. As part of a study on the role of cytokine and hematopoietic growth factors in central nervous system (CNS) development and inflammation, we examined the expression of CSF-1 in dissociated glial cells cultured from human fetal CNS tissue. CSF-1 mRNA and protein were constitutively expressed by astrocytes. The steady state level of CSF-1 mRNA was markedly up-regulated by both IL-1 beta and TNF-alpha in a time- and dose-dependent manner, whereas only a minimal increase was detected after stimulation with LPS. In unstimulated astrocyte cultures, CSF-1 protein levels gradually increased to 3.5-fold base-line values by 96 h and were significantly increased by all three stimulants in the order of IL-1 > or = TNF > LPS. Low levels of CSF-1 mRNA and protein were also detected in unstimulated microglia cultures. In contrast to astrocyte cultures, CSF-1 mRNA and protein increased significantly after stimulation with LPS, but changed only minimally after exposure to TNF-alpha or IL-1 beta. The effect of CSF-1 on cell proliferation, morphology, and class II MHC Ag expression was determined in highly enriched cultures of microglia and astrocytes. Microglia treated with CSF-1 showed a modest level of proliferation and differentiation into rod-shaped cells, whereas neither cell number nor shape was changed in astrocyte cultures. Interestingly, marked inhibition of both basal and IFN gamma-induced class II MHC Ag expression was observed in microglial cells cultured in the presence of CSF-1, whereas no effect was detected in astrocytes. These results suggest the possibility that in situ production of CSF-1 in the CNS may regulate normal glial cell development and contribute to the immunologic status of the CNS through the down-regulation of class II MHC expression. PMID- 8419492 TI - Stimulation of tyrosine phosphorylation and calcium mobilization by Fc gamma receptor cross-linking. Regulation by the phosphotyrosine phosphatase CD45. AB - The binding and subsequent cross-linking of murine IgG2a or human IgG to the Fc gamma R on the monocytic cell line THP-1 induced a rapid, dose-dependent increase in tyrosine phosphorylation of several proteins (a doublet centered around 110 kDa, and bands at 80, 60, and 52 kDa) and smaller increases in other proteins. This phosphorylation was accompanied by an increase in intracellular free Ca2+. The signaling required the cross-linking of the IgG, either through a biotin avidin complex or with a F(ab')2 second antibody. Cross-linking of an F(ab')2 fragment of mAb 32.2 to Fc gamma RI (CD64) or an Fab fragment of mAb IV.3 to Fc gamma RII (CDw32) gave similar results to those observed with intact murine IgG2a or human IgG. Cross-linking of a F(ab')2 fragment of mAb 3G8 to Fc gamma RIII (CD16) had very little effect. Increases in both tyrosine phosphorylation and intracellular free Ca2+ were significantly reduced in a dose-dependent manner upon treatment of THP-1 cells with the tyrosine kinase inhibitors herbimycin-A, genistein, or erbstatin. Additionally, there was a marked inhibition of both Ca2+ mobilization and tyrosine phosphorylation when a F(ab')2 fragment of a mAb (T191) to the protein tyrosine phosphatase CD45, was co-cross-linked with either Hu-IgG, Mu-IgG2a, F(ab')2 anti-Fc gamma RI, or Fab anti-Fc gamma RII. Taken together these results suggest that signaling through Fc gamma RI (CD64) and Fc gamma RII (CDw32) in the monocytic leukemia cell line THP-1 gives rise to rapid tyrosine phosphorylation of several proteins followed by an increase in intracellular calcium. In addition, CD45 is able to inhibit the intracellular signaling when it is brought into close proximity to the Fc gamma R. This suggests that this transmembrane tyrosine phosphatase may regulate the stimulation of the cells through the Fc gamma R. PMID- 8419493 TI - Studies of T cell deletion and T cell anergy following in vivo administration of SEB to normal and lupus-prone mice. AB - This study examines the responses of lupus-prone NZB, (NZB x NZW) F1, BXSB, MRL lpr/lpr and control mice (H-2 and Mls matched) to in vivo administration of the superantigen staphylococcal enterotoxin B (SEB). Two weeks after i.v. administration of 500 micrograms SEB, CD4+V beta 8+ lymph node T cells were deleted equivalently by lupus-prone and control mice. However, IE+ strains deleted a greater proportion (47% to 77%) of their CD4+V beta 8+ cells than did IE- strains (24% to 27%). CD8+V beta 8+ cells were deleted less than CD4+V beta 8+ cells by injection of 500 micrograms SEB. IE- strains failed to delete CD8+V beta 8+ cells, whereas six of seven IE+ strains deleted > 25% of their CD8+V beta 8+ cells. IE+ MRL-lpr/lpr mice showed some impairment in deletion: they failed to delete CD8+V beta 8+ cells at all doses of SEB and had reduced deletion of CD4+V beta 8+ cells at low doses of in vivo SEB (10 and 50 micrograms). Peripheral expansion of the intrathymically deleted V beta 7 TCR family was not observed in lupus-prone mice 2 wk after 500 micrograms in vivo SEB. In vitro restimulation with SEB of mice previously injected with 500 micrograms SEB demonstrated anergy in T cells from all strains, including the IE- and MRL-lpr/lpr. This result contrasts with previous reports of tolerance defects in lupus-prone strains using B cell read-out assays as measures of tolerance. The present study demonstrates that there is no global defect in peripheral T cell deletion or anergy in lupus prone mice to the superantigen SEB. Although additional Ag would need to be studied, these experiments raise the possibility that some reported tolerance defects in lupus-prone strains may reflect excessive B cell responses to relatively normal T cell signals. PMID- 8419494 TI - MRL mice produce anti-Su autoantibody, a specificity associated with systemic lupus erythematosus. AB - Certain autoimmune mouse strains exhibit features similar to human SLE. To discover genetic and immunologic events governing expression of a new SLE associated antibody, the presence of anti-Su and its relationship to other SLE related antibodies (anti-Sm, -ribonucleoprotein, -Ro (SS-A), -La (SS-B)) were determined in MRL and other autoimmune and nonautoimmune mice. By double immunodiffusion, sera from 34/183 (19%) 4- to 10-mo-old MRL/Mp-lpr/lpr (MRL/lpr) and 28/108 (26%) 8- to 20-mo-old MRL/Mp(-)+/+ (MRL/+) mice were positive for anti Su antibodies. Anti-Sm antibodies were found in 60/183 (33%) and 39/108 (36%) of these animals, respectively. The two specificities were found together in individual mice more frequently than would be predicted by chance. In contrast, C57BL/6-lpr/lpr (B6/lpr) mice rarely showed either specificity. Analysis of F1 hybrids between B6/lpr and MRL/lpr and of F1 x MRL/lpr backcross mice suggested that a small number of autosomal recessive genes controlled the anti-Su and anti Sm responses. With the exception of a single NZB serum sample, NZB, BXSB, and nonautoimmune mice were negative for all antibodies tested, and no mice were positive for anti-RNP, anti-Ro, or anti-La. MRL/lpr and MRL/+ autoimmune mice thus provide unique models for human SLE, because they express several of the SLE specific marker autoantibodies. These models should be useful in disclosing molecular and immunologic events governing autoantibody expression in this condition. PMID- 8419495 TI - Vasectomy procedure. PMID- 8419496 TI - Human Genome Project. PMID- 8419497 TI - Lovastatin vs PIB. PMID- 8419498 TI - One in a million: when extraordinary cases occur in an ordinary practice. PMID- 8419499 TI - Cards, cakes, and homegrown tomatoes. PMID- 8419500 TI - Glucose intolerance as a consequence of oral terbutaline treatment for preterm labor. AB - BACKGROUND: In this case series, glucose regulation is examined prospectively during treatment with terbutaline sulfate for premature labor in women who were (1) previously documented as nondiabetic, (2) found to have gestational diabetes mellitus (GDM), and (3) tested for glucose intolerance while terbutaline was being administered. The glucose profiles of women treated with terbutaline were contrasted with the profiles of nondiabetic women and women known to have GDM who were not in premature labor. METHODS: Subjects tested capillary blood glucose an average of five times a day during terbutaline treatment and for 1 week after terbutaline treatment was discontinued. They used memory-based reflectometers that stored and transmitted self-monitored blood glucose data to a personal computer. RESULTS: A significant difference (P = .001) was found between average fasting glucose values (111 +/- 23 mg/dL) for the five nondiabetic subjects treated with terbutaline and values of patients in an historical control group (41 nondiabetic pregnancies [72 +/- 22 mg/dL]) who were not in premature labor. The four diabetic subjects and the one subject who had not been previously tested also experienced higher blood glucose levels during tocolytic therapy. Glucose levels returned to preintervention values with the cessation of terbutaline therapy. CONCLUSIONS: It has been previously suggested that terbutaline increases hepatic glycogenolysis, which may aggravate glucose intolerance. This phenomenon, combined with normal pregnancy-induced insulin resistance, may explain abnormal ambulatory glucose patterns in women who are euglycemic before introduction of terbutaline therapy. PMID- 8419501 TI - Physician awareness of prescription drug costs: a missing element of drug advertising and promotion. AB - BACKGROUND: Although the cost of prescription drugs is recognized as an important facet of health care expenditures, many physicians are purportedly unaware of actual drug costs. To test this hypothesis, we surveyed physician awareness of the cost of 20 actively marketed prescription drugs. METHODS: A questionnaire listing four possible cost categories for each drug was administered to 305 registrants of a 5-day family medicine continuing education course. RESULTS: Ninety-two physicians completed the questionnaire. Only one, a 40-year-old, board certified physician who had been in practice for 3 years, answered 70% of questions correctly. The average score for the other participants was 37% (range 0% to 75%). CONCLUSIONS: The majority of physicians questioned could not accurately identify the price range of commonly prescribed drugs. We recommend drug cost disclosure in drug advertising to help address this problem. PMID- 8419502 TI - A visit-based quantitative measure of family care. AB - Within the discipline of family medicine, providing family care receives varying emphasis. Studies are needed to more strongly link the process of family care to improvements in health outcomes. Such studies require validated measures with which to quantify the amount of care provided to families. This paper proposes a conceptual definition and an operational quantitative measure of family care. This family care measure, which we based on a three-dimensional operational model of family care (frequency of visits, number of family members, number of providers), translates a family's pattern of health care into an easily interpreted index. PMID- 8419503 TI - Effectiveness of an ethics consultation service. AB - BACKGROUND: Ethics consultation is a relatively new service in clinical medicine. Most such services have been developed in departments of internal medicine. Few studies have evaluated the results of such consultations, and none have examined whether a family practice perspective enhances the consultation process. METHODS: An ethics consultation service was established in the Department of Family Medicine at Loma Linda University School of Medicine in 1990. Data were collected from the consultations performed during the first year. A questionnaire was sent to the attending physicians for their evaluation of the service. RESULTS: Ethics consultations were provided to the health care teams of 46 patients in five clinical departments. The attending physicians found the consultations to be important in clarifying ethical issues, educating the team, increasing confidence in decisions, and in patient management in more than 90% of the cases; however, the consultations resulted in significant changes in patient management only 36% of the time. CONCLUSIONS: It is feasible to establish an ethics consultation service within a department of family medicine in a university hospital and to provide consultations to physicians in other specialties. PMID- 8419504 TI - Self-reported hearing difficulty and audiometric thresholds in nursing home residents. AB - BACKGROUND: Nursing home practitioners usually assess the general health and functional abilities of each resident at the time of admission. If audiometric screening is not routinely available, assessment of the resident's hearing status will probably consist only of asking questions about hearing difficulty. In this study we explored which questions, when answered positively, were most strongly associated with moderate or severe hearing impairment. METHODS: A total of 198 newly admitted nursing home residents answered questions regarding their hearing in common listening situations, and then underwent audiometric assessment. RESULTS: Fifty-four percent of the residents had a pure tone average hearing level of > 25 dB at 500, 1000, and 2000 Hz and were therefore considered impaired. A single general question regarding hearing had a sensitivity of 69% in determining the presence of such impairment. Three specific questions which assessed hearing--in a group, while watching television, and while on the telephone--had a collective sensitivity of 83%. Asking the three questions was significantly (P = .003) more effective than asking only the general question. CONCLUSIONS: A set of specific questions significantly improved the identification of residents whose hearing loss affected their daily living activities compared with the use of a single hearing loss question. PMID- 8419505 TI - Increasing compliance with mammography recommendations: health assessment forms. AB - BACKGROUND: Inexpensive reminder systems are needed to ensure that primary care physicians consistently provide health maintenance services to their patients. The purpose of this study was to determine the effectiveness of a simple, inexpensive health assessment form in place of the standard chart note to increase physician compliance with mammography recommendations. METHODS: A health assessment form with a reminder for screening mammography was implemented in a family practice in 1987 and was to be used as the official chart record for health maintenance visits. The charts of all women 50 years of age and older with two or more office visits during the years 1985 through 1988 were audited to determine how many mammograms were completed. Results were compared with mammography completion rates at a similar practice that did not use a health assessment form. RESULTS: The study group showed a significant increase in mammography completion after implementation of the form, with compliance increasing from 7.3% to 32.0% (P < .001). The comparison group had an increase in mammogram completion from 12.0% to 17.8% (P < .001). The difference between the changes in rates of mammography in the two practices was statistically significant (P < .001). Among women in the study group who had a scheduled health maintenance visit during the study period the average rate of mammography completion increased from 21.2% to 65.2% (P < .001). CONCLUSIONS: The addition of a health assessment form with a mammography reminder at the health maintenance visit is an effective and inexpensive method to increase compliance with mammography. PMID- 8419506 TI - A comparison of rural family practice in the 1930s and today. AB - Two hundred forty-four consecutive diagnoses and procedures appearing on the patient billing records between June 1934 and September 1935 of a general physician practicing in rural southwestern Minnesota were compared with 286 diagnoses and procedures taken from the billing records of patient visits made over a 2-week period to a modern family physician practicing in a comparable rural community in southwestern Ohio. The most common items on the billing records of the physician of the 1930s were follow-up incision and drainage of abscess, 26 (10.7%); diphtheria immunization, 24 (9.8%); follow-up drainage for mastoiditis, 17 (7.0%); and scrotal tap for epididymitis, 14 (5.7%). Many of these patient encounters were at the patient's home. The most common items on the records of the modern physician practicing in rural southwestern Ohio were upper respiratory tract infection, 13 (4.5%); hypertension, 12 (4.2%); hyperlipidemia, 11 (3.9%); and history-taking and physical examination (adult), 10 (3.5%). This study suggests that there are great differences between the diagnostic profiles of the first third of the 20th century and modern family physicians. Many of the common diagnoses seen by the physician of the 1930s required a procedure to be performed. Many of the problems treated by the contemporary family physician did not even exist for the early 20th century general physician. Some of the differences between the modern physician and his predecessor can be explained by the introduction of antibiotics in the late 1930s and early 1940s. PMID- 8419507 TI - Antihypertensive drug therapy and coronary heart disease risk. AB - Coronary heart disease (CHD) is the major cause of death in the United States. Major modifiable risk factors for CHD are hypertension, hypercholesterolemia, and cigarette smoking, with concomitant risk factors, especially left ventricular hypertrophy, that act synergistically to significantly increase overall risk. Antihypertensive therapy, while reducing the incidence of stroke, has not consistently reduced the incidence of CHD. This may be a result, in part, of adverse effects on the metabolic profile, especially on blood lipids, which are induced by diuretics and certain beta-blockers. Other antihypertensive agents appear to be either lipid neutral, such as calcium channel blockers and angiotensin-converting enzyme inhibitors, or lipid positive, such as selective alpha 1-blockers. The choice of initial antihypertensive therapy should be made with all of a patient's risk factors in mind. In addition to the drugs recommended in the 1988 Guidelines of the Joint National Committee on Detection, Evaluation and Treatment of High Blood Pressure, selective alpha 1-blockers should also be considered since they improve the lipid profile as well as reduce blood pressure. PMID- 8419508 TI - Iatrogenic diarrhea caused by sorbitol. AB - A case of severe diarrhea with devastating effects in a young patient with a head injury is reported. The cause was found to be sorbitol used as an inactive ingredient in a commonly prescribed medication. Sorbitol is present in many prescription and over-the-counter medications and should be considered in the differential diagnosis of patients who present with abdominal complaints. PMID- 8419509 TI - Pseudocyesis in an adolescent incest survivor. AB - This case is an example of pseudocyesis in an incest survivor. Symptoms of pregnancy were unconsciously created by the patient to shield her from the memory of her incest and at the same time confront the reality of her abuse. Her symptoms allowed her entrance into the medical system. Once in the system, it was just a matter of time before she was referred for psychological evaluation. The authors recommend that family physicians consider incest when evaluating patients who present with pseudocyesis. PMID- 8419510 TI - Information processing and social cognitive problem solving in schizophrenia. Assessment of interrelationships and changes over time. AB - The relationship between basic information processing and social cognitive problem solving (SCPS) was studied in 31 schizophrenic, 16 depressive, and 31 control subjects. The clinical subjects were assessed twice, during symptom exacerbation and 3 months later, after partial remission. Control subjects were tested during the same time period. Subjects completed a means-ends problem solving test, an alternative solution generation task, and an information processing test battery. Results showed that schizophrenic subjects demonstrated significant improvement in a number of information-processing indices over time. No significant changes on the social cognitive problem-solving variables were found. Furthermore, all three groups demonstrated different patterns of relationships between information processing and social cognitive problem solving. Implications for treatment are discussed. PMID- 8419511 TI - The concordance of operational criteria for mania and schizoaffective mania. AB - Indices of agreement of six definitions of mania and four definitions of schizoaffective mania were calculated in a series of 81 manically disturbed patients. In particular, the concordance of the manic triad definition of overactivity, pressure of speech, and manic mood with stricter definitions was measured. The agreement among criteria was generally low for mania and even lower for schizoaffective mania. Although the manic triad definition diagnoses a wider range of patients, its coverage is less than might be expected from the small number of symptoms included in the definition. Tighter constructs of mania generally do not seem much more valid. PMID- 8419512 TI - Reaction time crossover in schizotypal subjects. AB - The present study investigated the relationship between reaction time crossover and schizotypy as measured by Chapman's Physical Anhedonia Scale (PAS). Twenty PAS-identified schizotypal college students and 20 nonschizotypal students were administered a computerized version of Rodnick and Shakow's crossover reaction time task. The hypothesized interaction (crossover) between preparatory interval duration and preparatory interval presentation pattern was observed for PAS identified schizotypics. Results indicated that subjects identified on the basis of the PAS evidenced reaction time crossover patterns similar to those observed in schizophrenic and at-risk populations. PMID- 8419513 TI - The use of DSM and ICD diagnostic criteria in people with mental retardation. A review of empirical studies. AB - Empirical studies that have used DSM and ICD criteria with people who are mentally retarded were reviewed. They included both studies using clinical interviews and case notes for diagnosis and those using behavioral checklists. The merits of these methods, which have hitherto developed relatively independently, were critically evaluated. Areas for future development include: the application of the psychometric rigor, hitherto associated with behavioral checklists, to diagnosis based on clinical interviews and case notes; the development of broader and associated measures of the integrity of the diagnostic process; and the development and piloting of new criteria for some disorders for use with people with severe and profound mental retardation where current diagnostic criteria are inappropriate with this population. PMID- 8419514 TI - A longitudinal study of psychological distress in a cohort of gay men. Effects of social support and coping strategies. AB - This study presents analyses on questionnaire data collected from a panel of 520 gay men at risk for acquired immune deficiency syndrome, enrolled in the Coping and Change Study (1985-1987). The data were assessed to determine the association of social support and coping styles with subsequent depression and global distress and to investigate whether these predictors of mental health are stable or transient over time. Three different measures of the subjective, qualitative nature of social support were significantly associated with subsequent mental health. Those who reported a subjective sense of isolation experienced significantly more adverse mental health 6 months later at all three measurement periods. Scattered effects were found for perceived social conflict and perceived social support from others. These results indicate that certain types of social support appear to influence mental health in this cohort and, furthermore, that some associations are transient and others more stable over time. PMID- 8419515 TI - Time series analysis of intervention effects. Fluoxetine therapy as a case illustration. AB - This paper illustrates the advantages of time series analysis in documenting treatment effects through a case study of a trial of fluoxetine in a borderline woman being treated in a long-term inpatient unit for severe personality disorders. Data consisted of weekly self-reports of symptomatology over 58 weeks of hospitalization. Intervention analysis carried out after the patient was discharged documented the effectiveness of the medication and the differential timing of response in individual symptoms. PMID- 8419516 TI - Semantic structures and psychiatric diagnosis. AB - This study examines psychiatric diagnosis using the concepts and methods of cognitive anthropology. Clinicians are viewed as cultural experts whose knowledge is represented in semantic structures, which are arrangements of diagnostic categories based on similarity of meaning. A test of similarity judgments of diagnostic categories was administered to a group of clinicians to derive two semantic structures of the categories included in axis I and axis II, respectively, of DSM-III. These structures were then compared with actual clinical diagnoses of concurrent disorders, and with the expected incidence of such diagnoses, in order to determine the possible influence of semantic similarity on the diagnostic process. The results suggest that semantic meanings of diagnostic categories influence the use of rule-out diagnoses more than positive diagnoses, and that this influence is more pronounced with respect to axis II than axis I. PMID- 8419517 TI - Vocal alterations in schizophrenic speech. PMID- 8419518 TI - Can clozapine response be predicted? A naturalistic pilot study. PMID- 8419519 TI - Adult-onset stuttering as a presenting feature of schizophrenia: restoration of fluency with trifluoperazine. PMID- 8419520 TI - Dendritic reorganization of an identified neuron during metamorphosis of the moth Manduca sexta: the influence of interactions with the periphery. AB - During metamorphosis of the moth, Manduca sexta, an identified leg motor neuron, the femoral extensor motor neuron (FeExt MN) undergoes dramatic reorganization. Larval dendrites occupy two distinct regions of neuropil, one in the lateral leg neuropil and a second in dorsomedial neuropil. Adult dendrites occupy a greater volume of lateral leg neuropil but do not extend to the dorsomedial region of the ganglion. The adult dendritic morphology is acquired by extreme dendritic regression followed by extensive dendritic growth. Towards the end of larval life, MN dendrites begin to regress, but the most dramatic loss of dendrites occurs in the 3 days following pupation, such that only a few sparse dendrites are retained in the lateral region of leg neuropil. Extensive dendritic growth occurs over the subsequent days such that the MN acquires an adult-like morphology between 12 and 14 days after pupation. This basic process of dendritic remodeling is not dependent upon the presence of the adult leg, suggesting that neither contact with the new target muscle nor inputs from new leg sensory neurons are necessary for triggering dendritic changes. The final distribution of MN dendrites in the adult, however, is altered when the adult leg is absent, suggesting that cues from the adult leg are involved in directing or shaping the growth of MN dendrites to specific regions of neuropil. PMID- 8419521 TI - Effects of a protein synthesis inhibitor on the hormonally mediated regression and death of motoneurons in the tobacco hornworm, Manduca sexta. AB - The larval-pupal transformation of Manduca sexta is accompanied by the loss of the abdominal prolegs. The proleg muscles degenerate, the dendritic arbors of proleg motoneurons regress, and a subset of the proleg motoneurons dies. The regression and death of proleg motoneurons are triggered by the prepupal peak of ecdysteroids in the hemolymph. To investigate the possible involvement of protein synthesis in these events, we gave insects repeated injections of the protein synthesis inhibitor, cycloheximide (CHX), during the prepupal peak. Examination of insects 3-5 days following CHX treatment showed that CHX inhibited the death of proleg motoneurons and the production of pupal cuticle in a dose-dependent fashion. When insects were allowed to survive for 10 days after the final CHX injection, motoneuron death and pupal cuticle production sometimes occurred belatedly, apparently in response to the ecdysteroid rise that normally triggers adult development. CHX treatments that inhibited motoneuron death were less effective in inhibiting dendritic regression in the same neurons. In another set of experiments, abdomens were isolated from the ecdysteroid-secreting glands prior to the prepupal peak, and infused with 20-hydroxyecdysone (20-HE). Single injections of CHX delivered just prior to the start of the 20-HE infusion inhibited motoneuron death and pupal cuticle production, but in the range of doses tested, did not prevent dendritic regression. Our findings suggest that protein synthesis is a required step in the steroid-mediated death of proleg motoneurons, and that dendritic regression is less susceptible to inhibition by CHX than is motoneuron death. PMID- 8419522 TI - Rapid and protracted phases of retinal ganglion cell loss follow axotomy in the optic nerve of adult rats. AB - To investigate the short- and long-term effects of axotomy on the survival of central nervous system (CNS) neurons in adult rats, retinal ganglion cells (RGCs) were labelled retrogradely with the persistent marker diI and their axons interrupted in the optic nerve (ON) by intracranial crush 8 or 10 mm from the eye or intraorbital cut 0.5 or 3 mm from the eye. Labelled RGCs were counted in flat mounted retinas at intervals from 2 weeks to 20 months after axotomy. Two major patterns of RGC loss were observed: (1) an initial abrupt loss that was confined to the first 2 weeks after injury and was more severe when the ON was cut close to the eye; (2) a slower, persistent decline in RGC densities with one-half survival times that ranged from approximately 1 month after intraorbital ON cut to 6 months after intracranial ON crush. A small population of RGCs (approximately 5%) survived for as long as 20 months after intraorbital axotomy. The initial loss of axotomized RGCs presumably results from time-limited perturbations related to the position of the ON injury. A persistent lack of terminal connectivity between RGCs and their targets in the brain may contribute to the subsequent, more protracted RGC loss, but the differences between intraorbital cut and intracranial crush suggest that additional mechanisms are involved. It is unclear whether the various injury-related processes set in motion in both the ON and the retina exert random effects on all RGCs or act preferentially on subpopulations of these neurons. PMID- 8419523 TI - Behavior of fish retinal growth cones encountering chick caudal tectal membranes: a time-lapse study on growth cone collapse. AB - In a cross species in vitro assay, growth cones from fish temporal retina elongating on laminin lanes were observed with time-lapse videomicroscopy as they encountered lanes and territories that carried membrane fragments from the chick caudal tectum. Caudal tectal membranes of adult fish and embryonic chick are known to possess a repellent guiding component for temporal retinal axons. The caudal membranes of chick exert a particularly strong influence on fish temporal axons. Contacts with chick caudal membranes by just a few filopodia and parts of the lamellipodia evoked a turning response away from the membrane lane of the entire growth cone. Contacts by filo- and lamellipodia over the entire circumference of the growth cone, however, caused invariably growth cone collapse and retraction. During growth cone turning and collapse and retraction, filopodia remained in contact with the tectal membrane fragments, suggesting strong membrane-filopodia adhesion simultaneous to growth cone repulsion by the repellent guiding component. PMID- 8419524 TI - Ipsilateral retinopetal projection of the nucleus olfactoretinalis (NOR) during development and regeneration: a DiI study in a cichlid fish. AB - The development and regeneration of the ipsilateral retinopetal projection of the nucleus olfactoretinalis (NOR) in the cichlid fish Haplochromis burtoni was studied with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) in fixed tissue. Throughout development most NOR cells projected to the contralateral retina. Only an insignificant, transient elevation of a projection to the ipsilateral retina was found in a few animals; however, after severing the contralateral processes of NOR cells by either enucleation or nerve crush, many animals had significantly more NOR cells with a regenerated process to the ipsilateral retina. Nevertheless, within a few weeks of surgery, the number of animals with ipsilaterally projecting cells were reduced to control values. The transiently enhanced ipsilateral projections to the retina imply changes in the guiding mechanism after these operations and the existence of control mechanisms against unusual connections to the retina in this bony fish. PMID- 8419525 TI - Testosterone propionate administration prevents the loss of neurons within the central part of the medial preoptic nucleus. AB - In the rat, the central part of the medial preoptic nucleus (MPNc) of the male is larger in volume and has a greater number of neurons than that of the female. The nucleus of the female, however, can be "sex reversed" by exposing the rat to gonadal steroids perinatally. The purpose of the present study was to examine the development of the MPNc to determine when the sex difference first appears and whether this difference occurs due to the relative accumulation of neurons into the compact part of the MPNc of the male and sex-reversed female rat or to the loss of MPNc neurons in the control female. Pregnant, female Sprague-Dawley rats were given an injection of [3H]methyl thymidine on embryonic day 18 (E18). Rats were exposed to testosterone propionate (TP) or vehicle from E20 to postnatal day 10 (PN10) or until the time of sacrifice. Pups from three groups [males (oil), females (oil), and sex-reversed females (TP)] were sacrificed on PN2, PN4, PN7, PN10, or PN30. The volume of the compact part of the MPNc increased in males and sex-reversed females after PN4 but the volume in the nucleus of females remained relatively constant. The number of neurons and [3H]thymidine-labeled cells remained elevated from PN2-PN30 in males or sex-reversed females but decreased dramatically in oil-treated females between PN4 and PN7, reaching a minimal number by PN10. Cell cross-sectional area increased with age while cell density decreased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419526 TI - Hippocampal growth cone responses to focally applied electric fields. AB - A wide variety of cell types respond to electric fields in culture. Despite evidence for electric fields existing in the mammalian embryo, there are few studies testing the effects electric fields exert on neurons from the mammalian central nervous system (CNS). The present study demonstrates orientation responses to focally applied electric fields of embryonic rat hippocampal neurons isolated in culture. The most striking result from this study is that different growth cones of the same neuron can show differential responsiveness to focally applied electric fields: growth cones on the short, straight processes that are destined to become dendrites, oriented toward the cathode, whereas growth cones on the longest process, the presumptive axon, did not orient. The present experiments bring a significant increase in resolution to the study of neuronal growth cone orientation by applied electric fields: a novel examination of the early events leading to orientation. Growth cones on dendrites displayed a spectrum of orientation responses: directed lamellipodial extension, directed filopodial extension and/or reorientation, cytoplasmic swelling of existing filopodia, consolidation of filopodia, and rapid elongation of the entire process. Individual growth cones displayed only one or two of these responses. Additionally, not all growth cones on these short processes sustained their initial orientation response: 35% adapted within 6 min. PMID- 8419527 TI - Regulation of gamma-aminobutyric acid synthesis in the brain. AB - gamma-Aminobutyric acid (GABA) is synthesized in brain in at least two compartments, commonly called the transmitter and metabolic compartments, and because regulatory processes must serve the physiologic function of each compartment, the regulation of GABA synthesis presents a complex problem. Brain contains at least two molecular forms of glutamate decarboxylase (GAD), the principal synthetic enzyme for GABA. Two forms, termed GAD65 and GAD67, are the products of two genes and differ in sequence, molecular weight, interaction with the cofactor, pyridoxal 5'-phosphate (pyridoxal-P), and level of expression among brain regions. GAD65 appears to be localized in nerve terminals to a greater degree than GAD67, which appears to be more uniformly distributed throughout the cell. The interaction of GAD with pyridoxal-P is a major factor in the short-term regulation of GAD activity. At least 50% of GAD is present in brain as apoenzyme (GAD without bound cofactor; apoGAD), which serves as a reservoir of inactive GAD that can be drawn on when additional GABA synthesis is needed. A substantial majority of apoGAD in brain is accounted for by GAD65, but GAD67 also contributes to the pool of apoGAD. The apparent localization of GAD65 in nerve terminals and the large reserve of apoGAD65 suggest that GAD65 is specialized to respond to short-term changes in demand for transmitter GABA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419528 TI - Polyamine metabolism in experimental brain tumors of rat. AB - Biosynthesis and accumulation of the polyamines putrescine, spermidine, and spermine are closely associated with cellular growth processes. We examined polyamine levels and the activity of their first rate-limiting enzyme, ornithine decarboxylase (ODC), in stereotactically induced experimental gliomas of the rat brain 1 and 2 weeks after implantation. Regional ODC activity and polyamine levels were determined in the tumor and in the ipsi- and contralateral striatum, white matter, and cerebral cortex. In the tumor, both ODC activity and polyamine levels markedly increased with progressive tumor growth, as compared to those in the white matter of the opposite hemisphere. In the peritumoral brain tissue, ODC activity did not change, but there was a marked increase of putrescine and, to a lesser degree, of spermidine and spermine almost throughout the whole ipsilateral hemisphere. ODC activity, therefore, seems to be a reliable marker of neoplastic growth in the brain, which may be of use for new clinical concepts of the diagnosis and therapy of brain tumors. The more diffuse distribution of polyamines, however, may be associated with the formation and spreading of edema, which would explain some of the biological effects of tumors on distant brain tissue. PMID- 8419529 TI - A dystrophin-immunoreactive protein in mammalian brain. AB - Dystrophin is expressed only in muscle and brain, but is absent from all tissues of the adult mdx mouse, a mutant with a single base substitution in the dystrophin gene. The brains of both normal and mdx mice contain a protein of approximately 230 kDa that is recognised by anti-dystrophin antibodies raised to the N-terminal region of the rod-like domain. Although the N-terminal and central rod regions of dystrophin share structural homologies with spectrin, the 230-kDa protein represents neither of the presently described forms of brain spectrin by a variety of criteria (molecular weight, cerebellar localisation, and developmental regulation) and is distinct from the product of the dystrophin gene. Studies of mdx and normal mouse brain show different postnatal developmental regulation of the 230-kDa dystrophin-immunoreactive protein. PMID- 8419530 TI - Real-time monitoring of electrically stimulated norepinephrine release in rat thalamus: I. Resolution of transmitter and metabolite signal components. AB - Electrical stimulation of an ascending path of the locus ceruleus-norepinephrine system was used to elicit release of norepinephrine at noradrenergic terminal fields of the rat thalamus. Overflow into the extracellular fluid space was measured by fast in vivo chronoamperometry. At pretreated carbon fibers, the electrochemical signal consists of a sharp peak of approximately 20-30 s duration followed by a slower, plateau-like decay to baseline. The peak, characterized by a variety of pharmacological manipulations and dialysis perfusion, is primarily due to norepinephrine. The plateau was shown to correspond to metabolite efflux of 3,4-dihydroxy-phenylacetic acid. By varying the degree of electrochemical pretreatment, the response time and sensitivity of the fibers can be tuned to follow the entire signal or to select the separate components for detailed evaluation. This approach can be used to provide new information on the spatial and temporal characteristics of stimulated neurotransmitter release. PMID- 8419531 TI - Real-time monitoring of electrically stimulated norepinephrine release in rat thalamus: II. Modeling of release and reuptake characteristics of stimulated norepinephrine overflow. AB - As in the preceding study, electrical stimulation was used to effect release overflow of norepinephrine in the rat thalamus. Using a weak electrochemical pretreatment of a carbon fiber electrode, it was possible to "tune in" the electrochemical response signal for norepinephrine without metabolite interference. This reasonably selective signal was then used to study the degradation of norepinephrine release ability caused by prolonged stimulation. Further, the signals were modeled by the method used successfully for stimulated dopamine overflow, providing hitherto unavailable information on the temporal and spatial characteristics of norepinephrine release overflow. Pertinent comparisons between the release characteristics of the dopamine and norepinephrine systems show that the half-life for norepinephrine in the extracellular fluid space is approximately 1 s in thalamus compared with 33 ms for dopamine in caudate. PMID- 8419532 TI - Tau protein kinase II is involved in the regulation of the normal phosphorylation state of tau protein. AB - To study the phosphorylation state of tau in vivo, we have prepared antisera by immunizing rabbits with synthetic phosphopeptides containing phosphoamino acids at specific sites that are potential targets for tau protein kinase II. Immunoblot experiments using these antisera demonstrated that tau in microtubule associated proteins is phosphorylated at Ser144 and at Ser315. Almost all tau variants separated on two-dimensional gel electrophoresis were phosphorylated at Ser144 and nearly one-half of them at Ser315. Phosphorylation at Ser144 and at Thr147 of tau isolated from heat-stable brain extracts was shown to be developmentally regulated, with the highest level of phosphorylation found at postnatal week 1. In vitro phosphorylation of tau by tau protein kinase I, a kinase responsible for abnormal phosphorylation of tau found in paired helical filaments of patients with Alzheimer's disease, was enhanced by prior phosphorylation of tau by tau protein kinase II. Thus, we suggest that tau protein kinase II is indirectly involved, at least in part, in the regulation of the phosphorylation state of tau in neuronal cells. PMID- 8419533 TI - Comparison of [3H]WIN 35,428 binding, a marker for dopamine transporter, in embryonic mesencephalic neuronal cultures with striatal membranes of adult rats. AB - In contrast to striatal membranes of adult rats, where high- (KD1 = 34 nM) and low- (KD2 = 48,400 nM) affinity binding sites for [3H]WIN 35,428 are present, in primary cultures of ventral mesencephalon neurons (CVMNs) only low-affinity binding sites were found (KD = 336,000 nM). The binding of [3H]WIN 35,428 in CVMNs prepared from rat embryos was reversible, saturable, and located in cytosol. Although dopamine (DA) uptake blockers inhibited [3H]DA uptake at nanomolar concentrations in CVMNs, the displacement of [3H]WIN 35,428 binding in CVMNs by DA uptake inhibitors required 100-8,000 times higher concentrations than were needed to displace [3H]WIN 35,428 binding in striatal membranes. Piperazine derivatives, e.g., GBR-12909, GBR-12935, and rimcazole, inhibited [3H]WIN 35,428 binding in CVMNs more effectively than did cocaine, WIN 35,428, mazindol, nomifensine, or benztropin. A positive correlation (r = 0.779; p < 0.001) was found between drug affinities for the striatal membrane sites labeled by [3H]WIN 35,428 and their abilities to inhibit DA uptake in CVMNs, whereas no correlation existed between the IC50 values of drugs that inhibited [3H]WIN 35,428 binding and [3H]DA uptake in CVMNs. The cytosolic [3H]WIN 35,428 binding sites may be a piperazine acceptor and may not be involved in the regulation of the DA transporter. PMID- 8419534 TI - Amphetamine and other weak bases act to promote reverse transport of dopamine in ventral midbrain neurons. AB - Amphetamine-like psychostimulants are thought to produce rewarding effects by increasing dopamine levels at mesolimbic synapses. Paradoxically, dopamine uptake blockers, which generally increase extracellular dopamine, inhibit amphetamine induced dopamine overflow. This effect could be due to either inhibition of amphetamine uptake or inhibition of dopamine efflux through the transporter (reverse transport). We used weak bases and dopamine uptake blockers in ventral midbrain neuron cultures to separate the effects on blockade of amphetamine uptake from reverse transport of dopamine. Amphetamine, ammonium chloride, tributylamine, and monensin, at concentrations that produce similar reductions in acidic pH gradients, increased dopamine release. This effect was inhibited by uptake blockers. Although in the case of amphetamine the inhibition of release could have been due to blockade of amphetamine uptake, inhibition also occurred with weak bases that are not transporter substrates. This suggests that reduction of vesicular pH gradients increases cytoplasmic dopamine which in turn promotes reverse transport. Consistent with this model, extracellular 3,4 dihydroxyphenylacetic acid was increased by ammonium chloride and monensin, as would be expected with elevated cytoplasmic dopamine levels. These findings extend the weak base mechanism of amphetamine action, in which amphetamine reduces vesicular pH gradients resulting in increased cytoplasmic dopamine that promotes reverse transport. PMID- 8419535 TI - Laminin A, B1, and B2 chain gene expression in transected and regenerating nerves: regulation by axonal signals. AB - Laminin A, B1, and B2 chain mRNA levels in degenerating and regenerating mouse sciatic nerves were examined using northern blot analysis. In normal intact nerves, B1 and B2 mRNA steady-state levels were high, but when the nerves were crushed, the steady-state levels of B1 and B2 mRNA per milligram wet tissue weight of the distal segments of the nerves increased five- to eightfold over that of control levels as the total RNA and beta-actin mRNA levels increased, suggesting that these increases were the consequence of Schwann cell proliferation after axotomy. When the steady-state levels of B1 and B2 mRNA were normalized as the ratio to total RNA or beta-actin mRNA levels, however, they drastically decreased to about 20% of the normal nerve levels in the nerve segments distal to both the crush and transection sites 1 day after injury. In the crushed nerves, B1 and B2 mRNA levels gradually increased as the regenerating nerves arrived at the distal segments and reestablished normal axon-Schwann cell contact, and then returned to normal levels on the 21st day. In the transected nerves, where Schwann cells continued to be disconnected from axons, both B1 and B2 mRNA levels remained low. Cultured Schwann cells expressed detectable levels of B1 and B2 chain mRNA which significantly increased when the cells were cocultured with sensory neurons. However, mRNA for A chain was not detectable in the normal, axotomized nerves or in cultured Schwann cells. These data indicate that Schwann cells express laminin B1 and B2 chain mRNA that are up-regulated by axonal or neuronal contact, but they do not express A chain mRNA. PMID- 8419536 TI - Calcium-dependent and -independent acetylcholine release from electric organ synaptosomes by pardaxin: evidence of a biphasic action of an excitatory neurotoxin. AB - The effect of pardaxin, a new excitatory neurotoxin, on neurotransmitter release was tested using purely cholinergic synaptosomes of Torpedo marmorata electric organ. Pardaxin elicited the release of acetylcholine with a biphasic dose dependency. At low concentrations (up to 3 x 10(-7) M), the release was calcium dependent and synaptosomal structure was well preserved as revealed by electron microscopy and measurements of occluded lactate dehydrogenase activity. At concentrations from 3 x 10(-7) M to 10(-5) M, the pardaxin-induced release of acetylcholine was independent of extracellular calcium, and occluded synaptosomal lactate dehydrogenase activity was lowered, indicating a synaptosomal membrane perturbation. Electron microscopy of 10(-6) M pardaxin-treated synaptosomes revealed nerve terminals depleted of synaptic vesicles and containing cisternae. At higher toxin concentrations (> or = 10(-5) M), there were striking effects on synaptosomal morphology and occluded lactate dehydrogenase activity, suggesting a membrane lytic effect. We conclude that, at low concentrations, this neurotoxin is a promising tool to investigate calcium-dependent mechanisms of neurotransmitter release in the nervous system. PMID- 8419537 TI - Repeated reserpine increases striatal dopamine receptor and guanine nucleotide binding protein RNA. AB - In the present study the effects of repeated administration of reserpine on striatal dopamine receptor and guanine nucleotide binding protein mRNAs were determined. Twenty-four hours after seven consecutive daily injections of reserpine--a treatment that is known to produce functional sensitization of D1 and D2 dopamine receptors--the level of striatal D1 dopamine receptor mRNA was unchanged. However, the level of mRNA for the G protein Gs alpha was increased by 127%. After extended reserpine treatment for 14 days, levels of both striatal D1 DA receptor and Gs alpha mRNAs were elevated by 99 and 78%, respectively. Seven days of reserpine treatment also increased levels of mRNA of the striatal D2 dopamine receptor and of G proteins Gi2 alpha and Go alpha by 200, 79, and 32%, respectively. After 14 days of reserpine treatment the level of striatal D2 dopamine receptor mRNA was increased by twofold. In contrast, levels of the mRNAs coding for the G proteins Gi2 alpha and Go alpha were unchanged. These data suggest that dopamine receptors and their respective G proteins play important roles in the development of sensitization of striatal dopamine receptors during repeated reserpine treatment. Furthermore, the persistent increase in level of striatal Gs alpha mRNA suggests that this G protein is necessary to maintain supersensitivity of the striatal D1 dopamine receptor system following long-term dopamine depletion. PMID- 8419538 TI - Ca2+ release from inositol 1,4,5-trisphosphate-sensitive Ca2+ store by antidepressant drugs in cultured neurons of rat frontal cortex. AB - The ability of antidepressant drugs (ADs) to increase the concentration of intracellular Ca2+ ([Ca2+]i) was examined in primary cultured neurons from rat frontal cortices using the Ca(2+)-sensitive fluorescent indicator fura-2. Amitriptyline, imipramine, desipramine, and mianserin elicited transient increases in [Ca2+]i in a concentration-dependent manner (100 microM to 1 mM). These four AD-induced [Ca2+]i increases were not altered by the absence of external Ca2+ or by the presence of La3+ (30 microM), suggesting that these ADs provoked intracellular Ca2+ mobilization rather than Ca2+ influx. All four ADs increased inositol 1,4,5-trisphosphate (IP3) contents by 20-60% in the cultured cells. The potency of the IP3 production by these ADs closely correlated with the AD-induced [Ca2+]i responses. Pretreatment with neomycin, an inhibitor of IP3 generation, significantly inhibited amitriptyline- and imipramine-induced [Ca2+]i increases. In addition, by initially perfusing with bradykinin (10 microM) or acetylcholine (10 microM), which can stimulate the IP3 generation and mobilize the intracellular Ca2+, the amitriptyline responses were decreased by 76% and 69%, respectively. The amitriptyline-induced [Ca2+]i increases were unaffected by treatment with pertussis toxin. We conclude that high concentrations of amitriptyline and three other ADs mobilize Ca2+ from IP3-sensitive Ca2+ stores and that the responses are pertussis toxin-insensitive. However, it seems unlikely that the effects requiring high concentrations of ADs are related to the therapeutic action. PMID- 8419539 TI - Effect of P2Y agonists on adenosine transport in cultured chromaffin cells. AB - Adenosine transport in cultured chromaffin cells was inhibited by purinergic P2y receptor agonists without significant changes in the affinity constant, the values being between 1 +/- 0.4 and 1.6 +/- 0.6 microM. The Vmax parameter was modified significantly, being 40 +/- 1.0, 26 +/- 5.0, 32 +/- 3.0, and 22 +/- 4.7 pmol/10(6) cells/min for control, adenosine-5'-O-(2-thiodiphosphate), 5' adenylylimidodiphosphate, and P1,P4-di(adenosine-5'-) tetraphosphate (Ap4A) (100 microM for every effector), respectively. Ap4A, a physiological ligand for P2y receptors in chromaffin cells, showed the highest inhibitory effect (45%). This transport inhibition is explained by an increase in the cytosolic Ca2+ concentration ([Ca2+]i) and the activation of protein kinase C (PKC). Experiments of [Ca2+]i measurement with the fura-2 technique showed that P2y agonists, as well as bradykinin, were able to increase [Ca2+]i, this effect being independent of the presence of extracellular Ca2+. The peptide bradykinin, determined to be coupled to phosphatidylinositol hydrolysis and internal Ca2+ mobilization in chromaffin cells, exhibited a behavior similar to that of P2y agonists in adenosine transport inhibition (39%). P2y agonists and bradykinin increased PKC activity associated with the membrane fraction (about 50% increase in particulate PKC activity with respect to controls). The present studies suggest that adenosine transport is regulated by P2y-purinergic receptors mediated via Ca2+ mobilization and PKC activation. PMID- 8419540 TI - Antisense GAP-43 inhibits the evoked release of dopamine from PC12 cells. AB - To investigate the role of the neuronal growth-associated protein GAP-43 (neuromodulin, B-50, F1, P-57) in neurotransmitter release, we transfected PC12 cells with a recombinant expression vector coding for antisense human GAP-43 cRNA. Two stable transfectants, designated AS1 and AS2, were selected that had integrated the recombinant sequence and expressed antisense GAP-43 RNA. Immunoblot analysis of proteins from AS1 and AS2 cells indicated that the level of GAP-43 in these cell lines was reduced. In the presence of extracellular calcium, a depolarizing concentration of K+ (56 mM) evoked dopamine release from control cells, but not from AS1 and AS2 cells. Similarly, the calcium ionophore A23187 evoked dopamine release from control cells, but was ineffective in stimulating dopamine release from AS1 and AS2 cells. The antisense transfectants, as well as the control cells, contained appreciable quantities of dopamine and secretory granules with a normal appearance. Because the expression of antisense GAP-43 RNA in PC12 cells leads to a decrease in GAP-43 expression and to the loss of evoked dopamine release, these results provide evidence of a role for GAP-43 in calcium-dependent neurotransmitter release. PMID- 8419541 TI - Support for radiolabeling of a glycine recognition domain on the N-methyl-D aspartate receptor ionophore complex by 5,7-[3H]dichlorokynurenate in rat brain. AB - Pretreatment with Triton X-100 more than doubled the binding of radiolabeled 5,7 dichlorokynurenic acid (DCKA), a proposed antagonist at a glycine (Gly) recognition domain on the N-methyl-D-aspartate (NMDA) receptor ionophore complex, in rat brain synaptic membranes. The binding exhibited an inverse temperature dependency, reversibility, and saturability, the binding sites consisting of a single component with a high affinity (27.5 nM) and a relatively low density (2.87 pmol/mg of protein). The binding of both [3H]DCKA and [3H]Gly was similarly displaced by numerous putative agonists and antagonists at the Gly domain in a concentration-dependent manner at a concentration range of 100 nM to 0.1 mM. Among the 24 putative ligands tested, DCKA was the second most potent displacer of the binding of both radioligands with no intrinsic affinity for the binding of [3H]kainic acid and alpha-amino-3-hydroxy-5- [3H]methylisoxazole-4-propionic acid (AMPA) to the non-NMDA receptors. In contrast, the other proposed potent Gly antagonist, 5,7-dinitroquinoxaline-2,3-dione, was active in displacing the binding of [3H]glutamic ([3H]Glu) and D,L-(E)-2-amino-4-[3H]propyl-5-phosphono-3 pentenoic acids to the NMDA recognition domain with a relatively high affinity for the non-NMDA receptors. In addition, the proposed antagonist at the AMPA sensitive receptor, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline, not only displaced weakly the binding of both [3H]-Gly and [3H]DCKA, but also inhibited the binding of (+)-5-[3H]methyl-10,11- dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine ([3H]MK-801) to an ion channel associated with the NMDA-sensitive receptor in the presence of added Glu alone in a manner sensitive to antagonism by further added Gly. Clear correlations were seen between potencies of the displacers to displace [3H]DCKA binding and [3H]Gly binding, in addition to between the potencies to displace [3H]-DCKA or [3H]Gly binding and to potentiate or inhibit [3H]MK-801 binding. All quinoxalines tested were invariably more potent displacers of [3H]DCKA binding than [3H]Gly binding, whereas kynurenines were similarly effective in displacing the binding of both [3H]Gly and [3H]DCKA. These results undoubtedly give support to the proposal that [3H]DCKA is one useful radioligand available in terms of its high selectivity and affinity for the Gly domain in the brain. Possible multiplicity of the Gly domain is suggested by the differential pharmacological profiles between the binding of [3H]Gly and [3H]DCKA. PMID- 8419542 TI - Cell specificity of molecular changes during memory storage. AB - The aeolid nudibranch, Hermissenda crassicornis, exhibits Pavlovian conditioning to paired light and rotational stimuli and it has been suggested that protein kinase C(PKC) may play a critical role in the cellular mechanism for this conditioned behavioral response in the B-cell photoreceptor. The present study was designed to further examine learning-specific PKC involvement in identified cellular areas, particularly those in the visual-vestibular network, of the Hermissenda nervous system after Pavlovian conditioning. As used in previous vertebrate studies, the highly specific PKC radioligand, [3H]phorbol-12,13 dibutyrate ([3H]-PDBU), was used to determine the binding characteristics of the molluscan protein receptor considered to be PKC. The binding was specific, saturable, and could be displaced by a soluble diacylglycerol analogue. The binding activity was distributed evenly between the cytosol and the membrane. All of these analyses suggest that [3H]PDBU binds primarily to PKC in Hermissenda as it does in many other systems. Computerized grain image analysis was then used to determine the cellular localization of PKC as a function of Pavlovian conditioning. The medial and intermediate B photoreceptor and the optic ganglion showed significantly increased [3H]PDBU binding in conditioned animals. The present results provide the first report of an associative learning change of a key signal transduction component in identified neurons. PMID- 8419543 TI - Ecto-protein kinase and surface protein phosphorylation in PC12 cells: interactions with nerve growth factor. AB - The phosphorylation of surface proteins by ecto-protein kinase has been proposed to play a role in mechanisms underlying neuronal differentiation and their responsiveness to nerve growth factor (NGF). PC12 clones represent an optimal model for investigating the mode of action of NGF in a homogeneous cell population. In the present study we obtained evidence that PC12 cells possess ecto-protein kinase and characterized the endogenous phosphorylation of its surface protein substrates. PC12 cells maintained in a chemically defined medium exhibited phosphorylation of proteins by [gamma-32P]ATP added to the medium at time points preceding the intracellular phosphorylation of proteins in cells labeled with 32Pi. This activity was abolished by adding apyrase or trypsin to the medium but was not sensitive to addition of an excess of unlabeled Pi. As also expected from ecto-protein kinase activity, PC12 cells catalyzed the phosphorylation of an exogenous protein substrate added to the medium, dephospho alpha-casein, and this activity competed with the endogenous phosphorylation for extracellular ATP. Based on these criteria, three protein components migrating in sodium dodecyl sulfate gels with apparent molecular weights of 105K, 39K, and 20K were identified as exclusive substrates of ecto-protein kinase in PC12 cells. Of the phosphate incorporated into these proteins from extracellular ATP, 75-87% was found in phosphothreonine. The phosphorylation of the 39K protein by ecto-protein kinase did not require Mg2+, implicating this activity in the previously demonstrated regulation of Ca(2+)-dependent, high-affinity norepinephrine uptake in PC12 cells by extracellular ATP. The protein kinase inhibitor K-252a inhibited both intra- and extracellular protein phosphorylation in intact PC12 cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419544 TI - Effects of chronic nicotine treatment on the accumulation of [3H]tetraphenylphosphonium by cerebral cortical synaptosomes. AB - Chronic exposure of rats to nicotine increases the number of [3H]nicotine binding sites in the brain; however, it is not clear whether nicotinic cholinergic receptor function is altered as well. In this study, we have used [3H]tetraphenylphosphonium as a probe of synaptosomal membrane potential to investigate whether exposure to nicotine in vivo alters the ability of cerebral cortical synaptosomes to maintain a potential difference and to depolarize in response to in vitro nicotine. Treatment of rats for 14 days with 0.475 mg of nicotine base/day via subcutaneously implanted minipumps resulted in a decrease in the synaptosomal accumulation of [3H]tetraphenylphosphonium in physiological buffer, corresponding to a decrease in estimated membrane potential from -55 mV to -50 mV. The onset of the decrease in membrane potential occurred after 7 days of in vivo nicotine treatment and was significantly correlated with an increase in [3H]nicotine binding to cerebral cortical synaptosomal (P2) membranes. Nicotine, at in vitro concentrations of 3-1,000 microM, decreased [3H]tetraphenylphosphonium accumulation in cerebral cortical synaptosomes from control animals. When compared to accumulation in buffer alone, in vitro nicotine and other nicotinic agonists did not significantly decrease [3H]tetraphenylphosphonium accumulation in cerebral cortical synaptosomes prepared from rats treated with nicotine in vivo. These studies provide evidence that chronic treatment with nicotine results in an average lower membrane potential in cerebral cortical synaptosomes and in functional down-regulation of the depolarization response to nicotinic cholinergic receptor stimulation. PMID- 8419545 TI - GT1b ganglioside prevents tetanus toxin-induced protein kinase C activation and down-regulation in the neonatal brain in vivo. AB - A single dose of 0.25 ng of tetanus toxin (TeTx), equivalent to approximately 5 minimal lethal doses, injected intracerebrally to 1-day-old rats, caused translocation, i.e., activation, of Ca(2+)-phosphatidylserine-dependent protein kinase C (PKC) from the cytosolic to the membrane compartment within 1 h. Six hours after treatment with the toxin, a 40-50% reduction in the total brain PKC (cytosolic plus membrane) activity was noticed. GT1b (2 micrograms per brain) ganglioside, a putative receptor for TeTx, completely prevented enzyme translocation when injected intracerebrally 30 min before toxin administration and abolished down-regulation after 6 h from the time of toxin injection. GM1 (2 micrograms per brain), a ganglioside of lesser affinity for TeTx, produced by itself a 20-30% reduction of the total PKC activity and did not reverse TeTx induced PKC down-regulation after 6 h. 12-O-Tetradecanoylphorbol 13-acetate (TPA) phorbol ester, administered at a concentration of 5 x 10(-5) M, caused activation and down-regulation of the enzyme, although with several orders of magnitude lesser potency. GT1b prevented the TPA-induced down-regulation. PMID- 8419546 TI - Effects of increased gamma-aminobutyric acid levels on GAD67 protein and mRNA levels in rat cerebral cortex. AB - Rats were injected with saline or the gamma-aminobutyric acid (GABA) transaminase inhibitor gamma-vinyl-GABA for 7 days and the effects on GABA content and glutamic acid decarboxylase (GAD) activity, and the protein and mRNA levels of the two forms of GAD (GAD67 and GAD65) in the cerebral cortex were studied. gamma Vinyl-GABA induced a 2.3-fold increase in GABA content, whereas total GAD activity decreased by 30%. Quantitative immunoblotting showed that the decline in GAD activity was attributable to a 75-80% decrease in GAD67 levels, whereas the levels of GAD65 remained unchanged. RNA slot-blotting with a 32P-labeled GAD67 cDNA probe demonstrated that the change in GAD67 protein content was not associated with a change in GAD67 mRNA levels. Our results suggest that GABA specifically controls the level of GAD67 protein. This effect may be mediated by a decreased translation of the GAD67 mRNA and/or a change in the stability of the GAD67 protein. PMID- 8419547 TI - Pharmacological and regional characterization of [3H]LY278584 binding sites in human brain. AB - Binding of [3H]LY278584, which has been previously shown to label 5 hydroxytryptamine3 (5-HT3) receptors in rat cortex, was studied in human brain. Saturation experiments revealed a homogeneous population of saturable binding sites in amygdala (KD = 3.08 +/- 0.67 nM, Bmax = 11.86 +/- 1.87 fmol/mg of protein) as well as in hippocampus, caudate, and putamen. Specific binding was also high in nucleus accumbens and entorhinal cortex. Specific binding was negligible in neocortical areas. Kinetic studies conducted in human hippocampus revealed a Kon of 0.025 +/- 0.009 nM-1 min-1 and a Koff of 0.010 +/- 0.002 min-1. The kinetics of [3H]LY278584 binding were similar in the caudate. Pharmacological characterization of [3H]-LY278584 specific binding in caudate and amygdala indicated the compound was binding to 5-HT3 receptors. We conclude that 5-HT3 receptors labeled by [3H]LY278584 are present in both limbic and striatal areas in human brain, suggesting that 5-HT3 receptor antagonists may be able to influence the dopamine system in humans, similarly to their effects in rodent studies. PMID- 8419548 TI - Phosphatidylethanol affects inositol 1,4,5-trisphosphate levels in NG108-15 neuroblastoma x glioma hybrid cells. AB - Phosphatidylethanol is formed by phospholipase D in animal cells exposed to ethanol. Previous reports have demonstrated that the degradation of phosphatidylethanol is slow, indicating that this lipid may be present in the cells after ethanol itself has disappeared. Accumulation of an abnormal alcohol metabolite may influence cellular functions. In the present study, cultivation of NG108-15 neuroblastoma x glioma hybrid cells in the presence of ethanol resulted in an accumulation of phosphatidylethanol and a simultaneous increase in basal inositol 1,4,5-trisphosphate levels. The direct effects of phosphatidylethanol on the phosphoinositide signal transduction system were examined through incorporation of exogenous phosphatidylethanol into membranes of ethanol-naive cells. An incorporation amounting to 2.8% of cellular phospholipids was achieved after a 5-h incubation with 30 microM phosphatidylethanol. Phosphatidylethanol was found to cause a time- and dose-dependent increase in the basal levels of inositol 1,4,5-trisphosphate. The effects on inositol 1,4,5-trisphosphate levels of exogenously added phosphatidylethanol and ethanol exposure for 2 days were not additive. No effect on bradykinin-stimulated inositol 1,4,5-trisphosphate production could be detected. However, the increase in basal inositol 1,4,5 trisphosphate levels indicates that phosphatidylethanol affects inositol 1,4,5 trisphosphate turnover and emphasizes the importance of considering phosphatidylethanol as a possible mediator of ethanol-induced effects on cellular processes. PMID- 8419549 TI - Characterization of membrane-bound and solubilized high-affinity binding sites for 5'-N-ethylcarboxamido[3H]adenosine from bovine cerebral cortex. AB - A high-affinity binding site for 5'-N-ethylcarboxamido[3H]adenosine ([3H]NECA) from bovine cerebral cortex has been characterized in its membrane-bound and solubilized state after gel filtration on Sepharose CL-6B. For detection of this site in membranes, it was necessary to remove metabolites with high affinities for this site enzymatically, e.g., adenosine by addition of adenosine deaminase and inosine by addition of nucleoside phosphorylase. The pore-forming peptide antibiotic alamethicin further enhanced binding of [3H]NECA to this site in membranes. In contrast to adenosine receptors and the adenotin-like low-affinity binding protein, this novel site was extremely sensitive against treatment with the sulfhydryl alkylating agent N-ethylmaleimide. In competition experiments, this site could be differentiated from adenosine receptors by its high affinity for adenine nucleotides and its lack of affinity for adenosine receptor antagonists. Inosine and its derivative S-(4-nitrobenzyl)-6-thioinosine were relatively potent ligands with Ki values in the high nano- and low micromolar range, respectively. We conclude that the high-affinity NECA binding site described previously in bovine striatum is not exclusively located in the striatum, but can also be detected in membrane preparations and soluble extracts of bovine brain cortex. PMID- 8419550 TI - Differential interaction of 1-methyl-4-phenylpyridinium ion with the putatively vesicular binding site of [3H]tyramine in dopaminergic and nondopaminergic brain regions. AB - The neurotoxin 1-methyl-4-phenylpyridinium ion (MPP+) in the brain striatum has recently been shown to bind at a putatively vesicular site labeled by [3H]tyramine ([3H]TY). Whereas in the rat and mouse striatum MPP+ antagonized TY binding competitively, in the cerebellum there was a mixed-type antagonism, which suggests the simultaneous occupancy of two different sites. Ki values from displacement curves revealed a fourfold difference in the affinity of MPP+ for TY sites in the two brain regions. The degeneration of central noradrenergic terminals induced by an intraperitoneal injection of the toxin N-(2-chloroethyl) N-ethyl-2- bromobenzylamine in rats decreased by 80% the maximal number of cerebellar TY binding sites, while not affecting striatal binding. Furthermore, guanethidine, a marker for noradrenaline (NA) vesicles, potently inhibited TY binding in NA-innervated regions, such as the cerebellum and the parietal cortex, and poorly in the striatum. It is concluded (a) that both MPP+ and TY may also label NA vesicles and (b) that the vesicular carriers for dopamine and NA have different characteristics, which may underlie a regional specificity in the rate of endovesicular sequestration of MPP+, with either neurodegenerative or neuroprotective consequences, depending on the brain area involved. PMID- 8419551 TI - p-chlorophenylalanine increases tryptophan-5-hydroxylase mRNA levels in the rat dorsal raphe: a time course study using in situ hybridization. AB - The effects of a single dose of p-chlorophenylalanine on the mRNA encoding tryptophan-5-hydroxylase (EC 1.14.16.4) in the rat dorsal raphe nucleus were analyzed using in situ hybridization. The levels of tryptophan-5-hydroxylase mRNA were markedly increased in cell bodies located in the ventromedial part of the dorsal raphe 1-2 days after p-chlorophenylalanine (300 mg/kg, i.p.) administration. This was followed by a decrease in the amount of tryptophan-5 hydroxylase mRNA, which returned to basal values by 5 days after treatment. An almost symmetric time course was observed for the midbrain serotonin concentration. Our results on the temporal pattern of changes in tryptophan-5 hydroxylase mRNA levels in the ventromedial part of the dorsal raphe are opposite to those reported for the enzyme activity and serotonin concentration after p chlorophenylalanine treatment. These changes may result from modifications in enzyme mRNA expression, suggesting that tryptophan-5-hydroxylase gene transcription is involved in feedback mechanisms regulating serotonin synthesis. PMID- 8419552 TI - Protection against acute MPTP-induced dopamine depletion in mice by adenosine A1 agonist. AB - The effects of the adenosine A1 agonist N6-cyclohexyladenosine (CHA) on MPTP induced dopamine (DA) depletion in the striatum of C57BL/6 mice were studied. Twenty hours after a single injection of MPTP (30 mg/kg, s.c.), the toxin caused 62% depletion of striatal DA. CHA (0.2-3 mg/kg, s.c.), when given together with MPTP, prevented the toxin-induced DA depletion in a dose-dependent manner. This protective action was apparently mediated by the A1 receptors, because this effect was selectively antagonized by pretreating the animals with the A1 antagonist 8-cyclopentyl-1,3-dipropylxanthine (25 mg/kg, i.p.) but not with the A2 antagonist 1,3-dipropyl-7-methylxanthine (25 mg/kg, i.p.). When CHA (3 mg/kg) was injected 5 h after MPTP administration, at which point striatal DA levels were already reduced significantly, a rapid and complete recovery of the striatal DA levels occurred. These neurochemical data suggest that the A1 agonist CHA is potentially useful as a neuroprotective agent against MPTP-induced toxicity. PMID- 8419553 TI - Serotonin 5-HT1A autoreceptor blockade potentiates the ability of the 5-HT reuptake inhibitor citalopram to increase nerve terminal output of 5-HT in vivo: a microdialysis study. AB - The present study addressed the possibility that disinhibition of serotonin (5 HT) autoreceptor-mediated negative feedback might potentiate the elevation of nerve terminal 5-HT output induced by selective 5-HT reuptake blockade. To this end, rats were given citalopram and the 5-HT autoreceptor-blocking agents (S)-UH 301 (5-HT1A) and (-)-penbutolol (5-HT1A/1B), and the effect on extracellular 5-HT in the ventral hippocampus was monitored by means of in vivo microdialysis. Citalopram (5 mg/kg, s.c.) approximately doubled the 5-HT output, a response that was markedly augmented by (S)-UH-301 (3 mg/kg, s.c.) and (-)-penbutolol (8 mg/kg, s.c.) and by combined treatment with (S)-UH-301 (3 mg/kg, s.c.) plus (-) penbutolol (1 microM; via the dialysis perfusion medium), but not by (-) penbutolol (1 microM) alone. These findings provide evidence that 5-HT, in particular 5-HT1A, autoreceptor-mediated negative feedback mechanisms are pivotal in determining the nerve terminal 5-HT output level after 5-HT reuptake inhibition. These findings have important implications for the interplay between different processes controlling 5-HT transmission in vivo and might possibly offer a lead toward novel, therapeutically exploitable principles. PMID- 8419554 TI - Endogenous D-serine in rat brain: N-methyl-D-aspartate receptor-related distribution and aging. AB - Recently, a substantial amount of free D-serine has been demonstrated in rat brain, although it has long been presumed that D-amino acids are uncommon in mammals. The anatomical distribution and age-related changes in endogenous D serine have been examined here to obtain insight into its physiological functions. Free D-serine exclusively occurs in brains, with a persistent high content from birth to at least 86 postnatal weeks. The patterns of the regional variations and the postnatal changes in brain D-serine are closely correlated with those of the N-methyl-D-aspartate (NMDA)-type excitatory amino acid receptor. Because D-serine potentiates NMDA receptor-mediated transmission by selective stimulation of the strychnine-insensitive glycine site of the NMDA receptor, it is proposed that D-serine is a novel candidate as an intrinsic ligand for the glycine site in mammalian brain. PMID- 8419555 TI - A randomized trial of bromocriptine for cocaine users presenting to the emergency department. AB - OBJECTIVE: To determine the practicality of treating heavy cocaine users with bromocriptine in an outpatient setting to reduce cocaine use and increase entry into drug treatment. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: VA Medical Center emergency department. PARTICIPANTS: Twenty-nine men presenting for minor medical complaints who used cocaine at least four times per week for the previous month. INTERVENTION: Subjects received either bromocriptine, 1.25 mg, or placebo three times per day for two weeks. MEASUREMENTS AND MAIN RESULTS: On three follow-up visits, subjects were tested for cocaine metabolites and surveyed about their drug use and symptoms. Eight of 14 subjects randomized to the bromocriptine group appeared for follow-up evaluation and seven enrolled in drug treatment. Three of these patients tested negative for cocaine at all three visits. Ten of 15 control patients appeared for follow-up visits and five enrolled in drug treatment. None of the control patients tested negative for cocaine for all three visits. There was no major side effect. CONCLUSION: Bromocriptine can be administered safely to heavy cocaine users in an emergency department setting. The question of whether bromocriptine will reduce the use of cocaine or promote entry into drug treatment deserves further research. PMID- 8419556 TI - Coming to terms with the era of AIDS: attitudes of physicians in U.S. residency programs. AB - PURPOSE: To examine factors associated with residents' willingness to provide care to persons with AIDS. PATIENTS AND METHODS: Survey of all senior residents in internal medicine and family medicine in ten geographically representative states in early 1989. RESULTS: Preferring not to care for persons with AIDS was less common in the western United States and more common among those with more conservative politics, men physicians, Asian physicians, and foreign medical graduates. Multivariate analysis revealed scores on six attitudinal scales (homophobia, dislike of intravenous drug users, professional responsibility, fear of AIDS, futility of providing AIDS care, and clinical difficulty of AIDS care) to be strong independent predictors of willingness to care for persons with AIDS (adjusted R2 = 0.42). The authors postulated a model in which these six fundamental attitudes functioned as intervening variables between demographic characteristics and expressed willingness to provide AIDS care. Regression results supported the hypothesis that the associations between demographic characteristics and willingness to provide AIDS care were mediated via these attitudes. However, having had ambulatory experience in AIDS care during residency was associated with future intentions to provide such care, independent of negative attitudes. CONCLUSIONS: Physician willingness to care for persons with AIDS is inversely related to fear of acquiring AIDS, viewing treatment as futile or difficult, dislike of certain risk groups, and a lower sense of professional responsibility. These results identify concerns and stresses that should be addressed by residency programs, and emphasize the need for attention to the ethics and ideals of the profession in medical education. PMID- 8419557 TI - Intern candidates who withdraw from contracts: incidence and internal medicine program directors' attitudes. AB - OBJECTIVE: To study the prevalence of intern candidates accepted through the National Resident Matching Program (NRMP) or after the match who subsequently withdraw and the attitudes of program directors regarding this issue. DESIGN: Mailed survey. SETTING: Four hundred fifty-four internal medicine program directors. MEASUREMENTS: Questionnaires assessing program directors' attitudes toward interns who contract for positions after having verbal or written agreements elsewhere (and the program directors who accept them), the importance of this problem, and the incidence of this problem. MAIN RESULTS: Fifty-five programs (of the 221 responding) experienced intern withdrawals. Programs with larger numbers of open positions after the match had more intern withdrawals (p = 0.03). Eleven of the program directors knew of the prior commitment of the intern, and in all cases the other program director was called for permission to accept the intern. Program directors had negative feelings about both the interns who withdrew and the program directors who accepted them. Community, municipal, and Veterans Affairs hospital program directors were significantly less negative than those in university and university-affiliated hospitals toward interns who withdrew from written commitments (p = 0.001) and the program directors who accepted them (p < 0.05). CONCLUSION: Problems with intern candidate withdrawals from offered/matched programs affect a significant proportion of programs, especially those with larger numbers of unmatched positions. Program directors are generally disapproving. PMID- 8419558 TI - The ethics objective structured clinical examination. AB - OBJECTIVE: To develop objective structured clinical examination (OSCE) stations to assess the ability of physicians to address selected clinical-ethical situations, and to evaluate inter-rater agreement in these stations. DESIGN: Two ten-minute OSCE stations were developed using video-taped encounters between attending physicians and standardized patients. One scenario involved a daughter requesting a do-not-resuscitate (DNR) order for her competent mother without the mother's knowledge; the other involved a competent elderly woman requesting not to be re-intubated if her congestive heart failure worsened. The scenarios were evaluated using foreign medical graduates taking an OSCE. Each candidate was scored on his or her interaction with a standardized patient in the two OSCE stations by two independent observers. PARTICIPANTS: Eight attending physicians from the Division of General Internal Medicine at the Toronto Hospital were used to develop the OSCE stations, and 69 foreign medical graduates taking the University of Toronto Pre-Internship Program OSCE were used to evaluate the stations. RESULTS: The inter-rater reliability coefficients for the DNR and intubation scenarios were 0.79 (95% CI 0.69-0.87) and 0.75 (95% CI 0.62-0.84), respectively. For the DNR station, the scores of the two examiners, on a scale of 0 to 10, agreed exactly for 34 candidates (50%), within one mark for 59 candidates (87%), and within two marks for 65 candidates (96%). For the intubation station, the scores of the two examiners agreed exactly for 27 candidates (40%), within one mark for 56 candidates (84%), and within two marks for 63 candidates (94%). CONCLUSIONS: The authors produced ethics OSCE stations with face and content validity and satisfactory inter-rater agreement. Ethics OSCE stations may be suitable for evaluating the ability of medical students and residents to address selected clinical-ethical situations. PMID- 8419559 TI - Description of a nurse practitioner inpatient service in a public teaching hospital. AB - A new nurse practitioner service at a public teaching hospital received patients from internal medicine teaching services. To determine the characteristics of the patients, the ratings by the residents, and the professional team costs, the authors performed a case-series study of the first 248 patients. Transferred patients had conditions that necessitated long hospitalizations, most frequently cerebrovascular accident, dementia, and pneumonia. Housestaff rated the service positively. Estimated professional costs were similar to teaching service costs after 15 months. The nurse practitioner inpatient service effectively cared for internal medicine patients with long lengths of stay and received favorable housestaff ratings. PMID- 8419560 TI - Changing the fourth-year medicine clerkship structure: a successful model for a teaching service without housestaff. AB - The fourth-year medicine clerkship was restructured at one of four sites. Teams consisting of one attending physician and three students cared for eight new patients per week. The overall experience of the restructured clerkship was rated significantly more positively than the experiences at all other sites and the teaching effectiveness of the faculty more positively than those at two of the other sites. This clerkship structure was perceived by students to be a positive experience. It could serve as a model for housestaff training programs to successfully care for patients without housestaff. PMID- 8419561 TI - Lithium intoxication: a coordinated treatment approach. AB - This case illustrates the clinical features of lithium intoxication and the problems in treating it that may arise as a result of lithium's effects on the kidney. It also demonstrates the difficulties that can develop when a delicate physiologic balance is inadvertently disrupted by nonpharmacologic interventions such as seclusion and consequent restriction of access to food and water. Patients with lithium-induced urine-concentrating defects are especially at risk for dehydration, and care must be taken to ensure adequate fluid and salt intake. This case also shows how intense negative feelings evoked by chronically mentally ill patients can adversely affect their psychiatric and medical care. While such feelings are inevitable, their impact may be lessened by improved communication and coordination between the medical and psychiatric systems of care and by the presence of psychiatrists in the general medical hospital. PMID- 8419563 TI - Treating drug abuse: beyond bromocriptine. PMID- 8419562 TI - Primary care of lesbian patients. PMID- 8419564 TI - Ethical issues in resident recruitment. PMID- 8419565 TI - Assessing competence in clinical ethics: are we measuring the right behaviors. PMID- 8419566 TI - Use of corticosteroids for aspiration pneumonitis. PMID- 8419567 TI - Simplified treatment of vascular lesions using the argon laser. AB - Treatment of mucosal lesions and abnormalities using the carbon dioxide laser is well documented in the dental literature; however, use of the argon laser for the ablation of intraoral vascular malformations is not. This article discusses the argon laser-soft tissue interaction and illustrates by case reports its application in the treatment of vascular lesions. PMID- 8419568 TI - Efficacy of temporomandibular joint arthroscopy: a retrospective study. AB - One hundred fifty temporomandibular joints (TMJs) in 109 patients underwent arthroscopic surgery with lysis, lavage, and debridement. The patients were evaluated for preoperative and postoperative interincisal opening, lateral and protrusive excursions, and pain. There was general improvement in all categories, with a reduction of pain to a level that was manageable in 93% of the patients. Temporomandibular arthroscopic surgery is an effective procedure, improving TMJ hypomobility and reducing pain. PMID- 8419569 TI - Modification of the maxillary Le Fort I osteotomy in cleft-orthognathic surgery: the bilateral cleft lip and palate deformity. AB - The adolescent with bilateral cleft lip and palate (BCLP) undergoing orthognathic surgery may have multiple residual clefting problems, including a mobile, dysplastic premaxilla and hypoplastic lateral maxillary segments, with each segment misaligned in three dimensions. These problems are commonly compounded by residual oronasal fistulas, bony defects, soft-tissue scarring from previous surgery, and the congenital absence of the maxillary lateral incisor teeth with resulting cleft-dental gaps. This article describes modifications of the Le Fort I osteotomy that allow for the simultaneous routine and safe management of these deformities. Results of this operation on 22 consecutive patients are reported, with findings of follow-up ranging from 1 to 5 years. The long-term parameters reviewed include closure of residual oronasal fistulas, stabilization of the premaxilla, cleft-dental gap closure, maintenance of attached gingiva at the cleft site, maintenance of a positive overjet and overbite, the need for prosthetics to complete dental rehabilitation, and surgical morbidity. PMID- 8419570 TI - Relationship of substance abuse to complications with mandibular fractures. AB - This retrospective study analyzed the relationship between complications and substance abuse following mandibular fracture. Over a 2-year period, the records of 352 patients with 589 mandibular fractures were reviewed for methods of treatment and other variables, including chronic abuse of drugs. An overall complication rate of 18.5% was found. Positive associations between complications and chronic abuse of alcohol and nonintravenous and intravenous drugs were found. Intravenous drug abusers had a 30%, nonintravenous drug abusers had a 19%, and chronic alcohol abusers had a 15.5% incidence of complications. Those individuals who did not use any drug chronically had a 6.2% complication rate. The results of this study show that chronic substance abuse can significantly affect treatment outcomes for management of mandibular fractures. PMID- 8419571 TI - A modified endaural approach to the temporomandibular joint. PMID- 8419572 TI - Mohs fixed-tissue excision of dentoalveolar bone in canines: a histologic evaluation. AB - Mohs surgery is a well-established surgical technique that involves conservative, microscopically guided excision of malignant lesions, thereby sparing the maximum amount of normal surrounding tissue. Current surgical therapy for oral cancer frequently involves resection with wide margins, resulting in significant cosmetic and functional deficits. This pilot study used the Mohs technique for in situ fixation and excision of maxillary and mandibular bone in mongrel dogs. Histologic examination of the excisional sites was carried out at 8 weeks to examine changes in the surrounding bone, periodontal ligament, and dental pulp of adjacent teeth. The findings suggest that zinc chloride fixative paste does not result in destruction of the dental pulp or surrounding dentoalveolar bone of teeth in the vicinity of a fixed-tissue excision. The Mohs fixed-tissue technique may allow preservation of oral structures that would otherwise be sacrificed in the presence of bony tumor involvement. PMID- 8419573 TI - Torque and pullout analysis of six currently available self-tapping and "emergency" screws. AB - Insertional torque (IT), stripping torque (ST), and uniaxial pullout tests were performed to evaluate the effectiveness of six screw systems (Wurzburg, Techmedica, Synthes, Timesh, Steinhouser, Luhr) in thin porcine rib. The Timesh 2.2-mm self-tapping screw produced the largest insertional and stripping torque of all systems tested as well as the largest difference between the insertional and stripping torque. The Timesh emergency screw also had the largest insertional and stripping torque as well as the largest difference between the insertional and stripping torque. In pullout tests, the Timesh screw was found to be the most retentive. The overall data indicated that the ideal self-tapping screw should have the largest difference possible between drill size and external diameter, a channel, and at least three self-tapping threads for maximum retention. PMID- 8419574 TI - Fractures of the mandibular condyle. PMID- 8419575 TI - A systematic approach to management of high-flow vascular malformations of the mandible. AB - High-flow vascular malformations of the mandible are associated with significant morbidity and mortality. As these lesions are rare, the experience that a clinician has with treatment may be limited. To facilitate understanding and treatment of such lesions in a controlled and efficient manner, a treatment protocol is presented that includes careful preoperative planning, embolization, and surgery. PMID- 8419576 TI - Progressive paresthesia of the trigeminal nerve. PMID- 8419577 TI - Carcinoma after enucleation of a calcifying odontogenic cyst: a case report. PMID- 8419578 TI - Temporal necrotizing infection of dental origin. PMID- 8419579 TI - The Le Fort I osteotomy as an approach to the midcranial base for tumor resection: case report. PMID- 8419580 TI - Intraoral tertiary syphilis (gumma) in a human immunodeficiency virus-positive man: a case report. PMID- 8419581 TI - Composite scalp strip graft for eyebrow reconstruction: case report. PMID- 8419582 TI - New surgical splint for segmental maxillary osteotomies. PMID- 8419583 TI - Surgical management of internal derangement of the temporomandibular joint. PMID- 8419584 TI - Orthognathic surgery: the correction of dentofacial deformities. AB - The field of orthognathic surgery offers ever-improving technology to better rehabilitate patients with dentofacial deformities. Hospital stays have been reduced due to improved surgery and anesthesia. Rigid internal fixation has increased comfort for many patients by eliminating the inconvenience of having the jaws wired together. Most important has been the realization that teamwork between the general dentist and the various specialty disciplines is indispensable for good patient care and the attainment of the very best results. PMID- 8419585 TI - Management of impacted teeth. PMID- 8419586 TI - Current concepts in management of facial trauma. AB - Open reduction and internal fixation have become routine in facial fracture treatment. Methodical evaluation and treatment planning should be based on the patient's injuries, extenuating factors, and the surgeon's experience. After initial treatment, patients with facial trauma may require orthodontic alignment of teeth, endodontic therapy, osteotomies, implants, vestibuloplasties, and scar revisions to achieve an acceptable functional and cosmetic result. The oral and maxillofacial surgeon is best qualified to coordinate and help deliver this interdisciplinary approach to treatment. PMID- 8419587 TI - Advances in microsurgical nerve repair. PMID- 8419588 TI - Surgical correction of craniofacial malformations. PMID- 8419589 TI - Current trends in implant reconstruction. AB - Significant advances have occurred in preprosthetic surgery in the past decade. Patients whose problems range from the loss of a single tooth to extreme conditions involving acquired or congenital defects now have options for reconstruction previously not possible. Optimal care most often requires a team approach with careful planning and execution. Ongoing basic research and strict clinical documentation involving current and future areas of treatment will provide an even greater degree of safety and effectiveness for our patients. PMID- 8419590 TI - Maxillofacial esthetic surgery. PMID- 8419591 TI - A team approach to the management of oral pathology. PMID- 8419592 TI - The changing epidemiology of cystic fibrosis. AB - Data from 17,857 patients with cystic fibrosis submitted in 1990 to the registry maintained by the Cystic Fibrosis Foundation were used to described their demographic characteristics, survival rates, pulmonary function, anthropometry, microbiologic data, complication rates, and health care utilization. Comparisons with similar data collected in 1969, 1972, and 1978 demonstrated a significant shift in the age distribution of patients with cystic fibrosis. The proportion of adult patients increased fourfold between 1969 (8%) and 1990 (33%). In 1990 the median age of all patients in the cystic fibrosis registry was 12.5 years; the median age at diagnosis was 7 months; cystic fibrosis was diagnosed in 90% of all patients by age 12 years. Meconium ileus at birth was reported for 16% of all patients with a new diagnosis in 1990. Median survival age doubled between 1969 and 1990, from 14 to 28 years. Female patients consistently had a lower median survival age than male patients (25 vs 30 years in 1990). The most frequently reported respiratory pathogen was Pseudomonas aeruginosa, cultured in specimens from 61% of all patients, ranging from 21% of those less than 1 year of age to more than 80% of those aged 26 years or older. Overall, patients with cystic fibrosis are living much longer than in the past but still have chronic pulmonary infections and other medical complications related to their disease, including diabetes, intestinal obstruction, cirrhosis, hemoptysis, and pneumothorax. PMID- 8419593 TI - Acute conjunctivitis in childhood. AB - We obtained specimens for culture from the lids and conjunctivae of 95 patients with acute conjunctivitis and 91 control children of similar age and, in addition, stained the conjunctival scrapings with Giemsa and Gram stains. The conjunctivitis was attributed to bacterial infection in 76 patients, viral infection in 12 children, and allergy in 2 patients; no cause was identified in the remaining 5 patients. In most cases the etiologic diagnosis was based on the results of laboratory studies. By separately culturing microorganisms in specimens from the lids and conjunctivae of patients and control subjects, we could distinguish normal flora from pathogens, and blepharitis from conjunctivitis. Staphylococci, corynebacteria, and alpha-hemolytic streptococci were the predominant organisms recovered from the lids of control subjects. In contrast, Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis were the major pathogens cultured from the conjunctival specimens from patients with bacterial conjunctivitis. Gram stains of conjunctival scrapings provided a rapid means of predicting the pathogen in 51 of 55 cases of bacterial conjunctivitis. Giemsa stains of conjunctival scrapings provided etiologic information in 81 of 84 cases, showing neutrophilia in bacterial infections, lymphocytosis in viral infections, and eosinophilia in allergic disease. These results indicate that most cases of acute conjunctivitis in children can be diagnosed on the basis of differential cultures of microorganisms from the lid and conjunctiva, together with Giemsa stains of conjunctival scrapings. PMID- 8419594 TI - Limitations of 3-phenylpropionylglycine in early screening for medium-chain acyl coenzyme A dehydrogenase deficiency. AB - Screening for medium-chain acyl-coenzyme A dehydrogenase (MCAD) deficiency by urinary 3-phenylpropionylglycine may not be reliable in early infancy because young infants are not colonized with adult-type colonic flora. In this study we delineated the microbes that produce 3-phenylpropionic acid, the precursor of 3 phenylpropionylglycine. We found that the use of some antibiotics may alter gut flora, thereby confounding this method of screening for MCAD deficiency. PMID- 8419595 TI - Hypovolemic shock and intestinal ischemia: a preventable complication of incomplete formula labeling. AB - A 19-day-old male infant was seen with full-thickness gangrene of the transverse colon. Hyperosmolar formula feedings caused by inadequate "directions for use" was the probable cause. Improved formula labeling might prevent further cases. PMID- 8419596 TI - Efficacy of venovenous extracorporeal membrane oxygenation for neonates with respiratory and circulatory compromise. AB - We report a 12-month experience at Egleston Children's Hospital in Atlanta, Ga., with a protocol under which venovenous extracorporeal membrane oxygenation (ECMO) was used instead of venoarterial ECMO. Fifty-five newborn infants were referred for ECMO, four of whom had disqualifying conditions (all four died). Thirty-one infants were supported without recourse to ECMO, one of whom died. Of the 20 remaining patients, three were placed on a venoarterial ECMO regimen because of our early uncertainty about the efficacy of venovenous ECMO or because of technical constraints. All other patients (n = 17), including three with congenital diaphragmatic hernia, were supported with venovenous perfusion. No patient begun on a venovenous ECMO regimen required conversion to venoarterial bypass. Before ECMO, venovenous patients required an average dopamine dose of 16 micrograms/kg per minute and an average dobutamine dose of 6 micrograms/kg per minute. Of 15 patients studied before ECMO, three had significantly impaired contractility, and all had evidence of pulmonary hypertension on an echocardiogram. Mean blood pressure did not change while heart rate fell from 172 to 146 beats/min during the first 2 hours of ECMO and vasoactive drug doses were reduced. Of the 17 venovenous ECMO patients, 15 (88%) survived. We conclude that neonatal patients with severe hypoxia and substantial circulatory compromise can be effectively supported by venovenous ECMO in most cases. PMID- 8419597 TI - Relationship between nutrition, weight change, and fluid compartments in preterm infants during the first week of life. AB - This study was conducted to investigate the redistribution of fluid compartments and to examine the factors contributing to the variability of early weight loss in premature infants. Fourteen preterm infants (mean +/- SD: birth weight, 1473 +/- 342 gm; gestational age, 30.7 +/- 2.4 weeks) were studied at 1 and 7 days of age. Total body water was measured by deuterium oxide dilution, extracellular volume by bromide dilution, and intracellular volume by the difference between total body water and extracellular volume. There were significant changes in body fluid distribution per concurrent weight from birth to age 1 week. Extracellular volume decreased by 11%, and intracellular volume increased by 8.5% with no change in total body water. Infants were then grouped according to postnatal weight loss (group 1 (n = 7) > 10% and group 2 (n = 7) < 5% of birth weight). In group 1 there was a significant loss of both weight (mean +/- SD: 15.6% +/- 3.7%) and extracellular volume (15.9% +/- 9% of birth weight), with no change in intracellular volume. In group 2 there was no significant weight loss (1.4% +/- 1.8%), but a significant loss of extracellular volume (13.0% +/- 5.4% of birth weight) and a significant increase in intracellular volume. Other differences between the groups were a lower energy intake in group 1 than in group 2 (mean +/ SD: 177 +/- 46 vs 269 +/- 45 kilojoules/kg per day; p < 0.005) and a higher physiologic stability index in group 1 (p < 0.05). We conclude that significant postnatal weight loss as a result of the contraction of the extracellular compartment occurs only in less stable infants whose energy intake is inadequate. With adequate energy intake, weight loss is minimal because of the expansion of the intracellular compartment, which may be related to the onset of growth. PMID- 8419598 TI - Diabetic fetal macrosomia: significance of disproportionate growth. AB - Fetal hyperinsulinism in infants of diabetic mothers (IDMs) produces increased fetal growth leading to macrosomia, which may or may not be proportionate. Disproportionate macrosomia refers to excessive weight characterized by a high weight/length ratio. We tested the hypotheses that (1) macrosomia in IDMs would be characterized by a high ponderal index (defined as weight/length ratio) and (2) infants with macrosomia who have a high ponderal index would have increased neonatal morbidity--specifically, hyperbilirubinemia, hypoglycemia, polycythemia, and acidosis. We studied 170 IDMs and 510 non-IDMs matched 1:3 for gestational age, race, and year of delivery. Forty-five percent of IDMs had macrosomia compared with 8% of control infants (p = 0.001), and 19% of IDMs had disproportionate macrosomia compared with 1% of control infants (p = 0.001). The rates of hyperbilirubinemia (p = 0.02), hypoglycemia (p = 0.01), and acidosis (p = 0.01) were greatest in infants with disproportionate macrosomia and least in nonmacrosomic infants. The incidence of polycythemia was not significantly different between the groups. We suggest that disproportionate macrosomia in the IDM is associated with an increased likelihood of neonatal complications. PMID- 8419599 TI - Outcome at twelve months of adjusted age in very low birth weight infants with lung immaturity: a randomized, placebo-controlled trial of human surfactant. AB - We compared the neurodevelopmental outcome of extremely premature, surfactant deficient infants who received either prophylactic surfactant at birth, "rescue" surfactant after the clinical diagnosis of respiratory distress syndrome was established, or placebo. Infants studied were participants in a randomized, bicenter (San Diego, Calif., and Helsinki, Finland), controlled trial of human surfactant therapy. One hundred fifty infants (prophylaxis group, 63 infants; rescue group, 57; placebo group, 30) were prospectively enrolled at 38 weeks of gestational age. There were no neonatal intergroup differences in the incidence or severity of sonographic central nervous system abnormality or retinopathy. One hundred forty-five infants were alive at 1 year of adjusted age, at which time growth, neurosensory, and neurologic outcome were similar in all three treatment groups at both centers. Cerebral palsy occurred in 20% overall. Five infants (3.5%) were functionally blind. However, infants treated at birth had lower mean mental and motor scores on the Bayley Scales of Infant Development compared with those of infants rescued with surfactant after the onset of respiratory distress syndrome (Mental Development Index: 78 vs 96, p = 0.02; Psychomotor Development Index: 73 vs 87, p = 0.04). Chronic lung disease occurred more frequently in the prophylactically treated group and contributed to the subjects' neurologic and developmental morbidity. Because prophylactic surfactant treatment offered no neurodevelopmental advantage and may contribute to poorer outcome, we currently recommend early surfactant replacement only for those infants who have postnatal evidence of respiratory distress syndrome. PMID- 8419600 TI - Helium-oxygen mixture in respiratory distress syndrome: a double-blind study. AB - In a randomized, controlled trial, the lungs of infants with respiratory distress syndrome were ventilated with either a helium-oxygen mixture or a nitrogen-oxygen mixture. In the helium-oxygen group, infants required a lower inspired oxygen concentration and a shorter duration of ventilation. There were also fewer deaths and fewer cases of bronchopulmonary dysplasia in the helium-oxygen group. PMID- 8419601 TI - Phase I evaluation of zidovudine administered to infants exposed at birth to the human immunodeficiency virus. AB - This study evaluated the safety, tolerability, and pharmacokinetics of zidovudine administered intravenously and orally to infants born to women infected with the human immunodeficiency virus. Thirty-two symptom-free infants were enrolled before 3 months of age. The pharmacokinetics of zidovudine were evaluated in each infant after single intravenously and orally administered doses of zidovudine on consecutive days, and during long-term oral administration of the drug for 4 to 6 weeks. As new patients were enrolled, doses of zidovudine were progressively increased from 2 to 4 mg/kg. Therapy was continued for up to 12 months in 7 of the infants proved to be infected with human immunodeficiency virus. Zidovudine was generally well tolerated; 20 children (62.5%) had anemia (hemoglobin level < 10.0 gm/dl) during therapy and 9 (28.1%) had neutropenia (neutrophil count < or = 750 cells/mm3); these hematologic abnormalities usually resolved spontaneously. The total body clearance of zidovudine increased significantly with age, from an average of 10.9 ml/min per kilogram in infants < or = 14 days of age to 19.0 ml/min per kilogram in older infants (p < 0.0001). Concurrently, there was a significant decrease in serum half-life from 3.12 hours in infants < or = 14 days to 1.87 hours in older infants (p = 0.0002). Oral absorption was satisfactory and bioavailability decreased significantly with age, from 89% in infants < or = 14 days to 61% in those > 14 days of age (p = 0.0002). Plasma concentrations of zidovudine were calculated to be in excess of 1 mumol/L (0.267 micrograms/ml) for 4.12 +/- 1.86 hours and 2.25 +/- 0.78 hours after oral doses of 2 mg/kg in infants younger than 2 weeks and 3 mg/kg in older infants, respectively. We conclude that zidovudine administered at oral doses of 2 mg/kg every 6 hours to infants aged less than 2 weeks and 3 mg/kg every 6 hours to infants older than 2 weeks resulted in plasma concentrations that are considered virustatic against human immunodeficiency virus. Zidovudine was well tolerated by infants at these doses. PMID- 8419602 TI - Effect of racemic epinephrine and salbutamol on clinical score and pulmonary mechanics in infants with bronchiolitis. AB - To test the efficacy of a combined alpha- and beta-receptor agonist in acute bronchiolitis, we compared inhaled racemic epinephrine with salbutamol in a double-blind, crossover, randomized protocol. Twenty-four infants, 4.6 +/- 0.5 (mean +/- SEM) months of age, with their first episode of bronchiolitis were tested. After sedation with chloral hydrate, a clinical score and pulmonary mechanics measurements using simultaneous signals of airflow volume and transpulmonary pressure were recorded. After baseline measurements, infants received either nebulized salbutamol, 0.03 ml/kg, or racemic epinephrine, 0.1 ml/kg. Thirty minutes later, there was a significant decrease in clinical score after treatment with racemic epinephrine compared with the baseline score (p < 0.001); this difference was not present after salbutamol inhalation (p = 0.42). Only 13 patients had a decrease in clinical score after salbutamol therapy, in comparison with 20 infants treated with racemic epinephrine (p < 0.01). Both drug decreased respiratory rate, but the decrease was greater after the use of racemic epinephrine (p < 0.001). There was a significant decrease in inspiratory, expiratory, and total pulmonary resistance after treatment with racemic epinephrine compared with baseline values (p < 0.01) but no significant change after salbutamol inhalation. There was no significant correlation between the clinical score and pulmonary mechanics either at baseline or after drug treatment. We conclude that racemic epinephrine is superior to salbutamol in the treatment of infants with their first episode of acute bronchiolitis. PMID- 8419603 TI - Gram-negative enteric bacillary meningitis: a twenty-one-year experience. AB - We reviewed our experience with gram-negative enteric bacillary meningitis in neonates and infants from 1969 through 1989. Ninety-eight patients were identified. Their ages were from 1 day to 2 years with a median of 10 days. In 25 patients (26%), predisposing factors were identified, the most common of which were neural tube defects and urinary tract anomalies. The causative agents were Escherichia coli (53%), Klebsiella-Enterobacter species (16%), Citrobacter diversus (9%), Salmonella species (9%), Proteus mirabilis (4%), Serratia marcescens (3%), Bacteroides fragilis (3%), and Aeromonas species (2%). At the time of diagnosis, Gram-stained smears of cerebrospinal fluid revealed gram negative bacilli in 61% of patients. The causative organism was cultured from blood obtained from 55% of patients, and 21% had positive urine culture results. The cerebrospinal fluid leukocyte counts ranged from 0 to 80,600 cells/mm3, and the cerebrospinal fluid/serum glucose concentration ratio was less than 0.5 in 72% of patients. Antimicrobial regimens varied greatly. After initiation of antibiotic therapy, an average of 3 days was needed for eradication of bacteria from cerebrospinal fluid. The case-fatality rate was 17%, and 61% of survivors had long-term sequelae that included seizure disorders, hydrocephalus, physical disability, developmental delay, and hearing loss. PMID- 8419604 TI - Adaptation of the meconium drug test for mass screening. AB - For purposes of mass drug screening, the procedure for analysis of meconium for drugs was modified into a one-step extraction and analysis by enzyme immunoassay. The accuracy of this modified method was tested by comparing the results of simultaneous analysis of 61 meconium samples for cocaine opiate and cannabinoid metabolites by both the original and the modified methods. In 61 samples analyzed, opiate was detected in 8 (13%) samples by the original method and in 9 (15%) by the modified method; cocaine was detected in 39 (64%) samples by the original method and in 39 (64%) by the modified method. The concordance between the negative and positive results of the modified versus the original methods was 98% and 100%, respectively, for opiate and 95% and 98%, respectively, for cocaine. Only one sample was positive for cannabinoid. Thus a comparison of positive results for cannabinoid was not done; however, all the negative results obtained by the modified method were confirmed by the original method. The clinical experience of mass meconium drug testing using the modified method in 1991 is also reported. In four centers tested (total tests = 4409), the prevalence of presence of drug was consistent with the high- or low-risk status of the population. This simplified, rapid procedure can be performed in most clinical laboratories. This adaptation has made the meconium drug test feasible for large-scale clinical and research use. PMID- 8419605 TI - James L. Gamble. PMID- 8419606 TI - Failure of intravenous pentamidine prophylaxis for Pneumocystis carinii pneumonia. PMID- 8419608 TI - Eponyms in medicine: possessive or nonpossessive? PMID- 8419607 TI - C-reactive protein and respiratory distress syndrome. PMID- 8419609 TI - Cerebral edema in maple syrup urine disease. PMID- 8419610 TI - Making a presumptive diagnosis of urinary tract infection by using a urinalysis performed in an on-site laboratory. AB - The purpose of our study was to characterize the utility of certain elements of urinalysis, singly or in combination, in identifying children with urinary tract infections (i.e., positive findings on urine culture). Laboratory results for urine specimens subjected to both urinalysis and culture in an on-site outpatient clinic (office) laboratory during a 36-month period were reviewed. All specimens were collected by one of three methods (the midstream void technique, urethral catheterization, or suprapubic aspiration of urine) from children with symptoms to assist in documenting a possible urinary tract infection. Specimen processing by certified technologists was initiated within 10 minutes of collection. Sensitivity, specificity, and positive and negative predictive values were determined for each test or combination of tests. Of 689 specimens, 102 (14.8%) had positive culture results. The combination of dipstick analysis (leukocyte esterase and nitrite tests) and of microscopic examination for bacteria had a sensitivity of 100% and a negative predictive value of 100%. The nitrite test had a specificity of 100% and a positive predictive value of 100%. We conclude that, when properly collected specimens are evaluated promptly by certified technologists, the rate of accuracy in detecting or ruling out a urinary tract infection (i.e., positive findings on urine culture) with certain elements of the urinalysis is high. PMID- 8419611 TI - Wegener granulomatosis in children and adolescents: clinical presentation and outcome. AB - We prospectively studied and compared clinical features, treatment, course of illness, and long-term morbidity and mortality rates for Wegener granulomatosis in 23 childhood-onset patients with those of 135 adult-onset patients who were studied concurrently. Treatment was usually provided with glucocorticoids and cyclophosphamide. The mean follow-up period was 8.7 years for childhood-onset and 7.6 years for adult-onset Wegener granulomatosis. Most aspects of Wegener granulomatosis were similar in childhood-onset and adult-onset patients. Permanent morbidity from disease occurred in 86% of both groups. However, some features were significantly different. Wegener granulomatosis in childhood-onset patients was complicated five times more often by subglottic stenosis and twice as often by nasal deformity. Treatment-related permanent morbidity occurred in 22% of childhood-onset patients and 45% of adult-onset patients. After similar periods of cyclophosphamide therapy and follow-up, cyclophosphamide-related malignancies were less likely (0% vs 11%) to have developed in childhood-onset patients. Although 89% of patients treated with glucocorticoids and cyclophosphamide had remission, prolonged delay in achieving remission and relapses led in both patient groups to freedom from active disease for approximately 50% of the total patient-years. As a result, morbidity was substantial and has led to comparative studies of alternative therapies. PMID- 8419612 TI - Effects of nightly clonidine administration on growth velocity in short children without growth hormone deficiency: a double-blind, placebo-controlled study. AB - Children with short stature, slowed linear growth velocity, and delayed skeletal maturation may secrete growth hormone (GH) normally in response to provocative stimuli but may also have spontaneous undersecretion of GH. Orally administered clonidine, an alpha 2-adrenergic agonist, is a potent acute stimulator of growth hormone releasing hormone-mediated pituitary GH release. We performed a double blind, placebo-controlled crossover study of nightly oral clonidine therapy (0.1 mg/m2) in 10 short, slowly growing, non-GH-deficient (stimulated GH level > 15 micrograms/L) prepubertal boys (range, 6.1 to 12.2 years; mean height standard deviation score, -2.3 +/- 0.4). Results of 6 months of clonidine therapy were compared with the results of 6 months of placebo therapy; GH responsiveness was subsequently assessed in 7 of 10 patients. Growth velocity (4.9 +/- 0.6 cm/yr baseline) was not improved by clonidine (4.6 +/- 1.2 cm/yr) or placebo (5.2 +/- 1.2 cm/yr), but it increased (p < 0.001) with GH therapy (8.2 +/- 1.3 cm/yr). Clonidine therapy similarly did not significantly affect plasma levels of insulin like growth factor I or bone age maturation. Diminution in clonidine-stimulated peak GH levels was not observed after long-term oral clonidine therapy. Thus, in contrast to previous non-placebo-controlled studies, nightly clonidine therapy did not increase growth velocity or plasma insulin-like growth factor I levels. Subsequent acceleration in growth velocity during GH therapy suggests that a proposed increase in clonidine-induced endogenous GH secretion does not result in an effective growth-promoting stimulus. PMID- 8419613 TI - Effects of long-term treatment with growth hormone on bone and mineral metabolism in children with growth hormone deficiency. AB - The effects of growth hormone (GH) deficiency and recombinant human GH replacement (0.6 IU/kg per week) on bone and mineral metabolism in 26 GH deficient children were studied for 12 months. Before therapy, all children had significantly reduced serum levels of osteocalcin, carboxyl-terminal propeptide of procollagen type I, and 1,25-dihydroxyvitamin D, whereas serum ionized calcium, phosphate, intact parathyroid hormone, calcitonin, and 25-hydroxyvitamin D concentrations were in the normal range. All children had significant reduction of bone density for their chronologic, statural, and bone ages. During therapy with recombinant human GH, a decrease of serum ionized calcium levels and increases of phosphate, osteocalcin, carboxyl-terminal propeptide of procollagen type I, and intact serum levels of parathyroid hormone were found. A significant increase of serum levels of 1,25-dihydroxyvitamin D was found at 12 months. The urinary phosphate/urinary creatinine ratio decreased, whereas values for nephrogenous cyclic adenosine monophosphate and the ratio of the maximum rate of renal tubular reabsorption of phosphate to the glomerular filtration rate increased. Bone density significantly improved at 12 months, with a complete recovery in 12 children (46.2%). Significant relationships were found among growth velocity, bone density, maximum tubular reabsorption/glomerular filtration rate ratio, and serum levels of carboxyl-terminal propeptide of type I procollagen. The changes in serum levels of this propeptide during the first week of recombinant human GH treatment were positively related to growth velocity at 6 and 12 months and to bone density at 12 months of treatment, whereas the changes in osteocalcin levels were not. We conclude that recombinant human GH treatment caused significant modifications of mineral metabolism and significantly increased bone density, and that measurement of serum levels of the propeptide during the first week of recombinant human GH administration may be a useful tool in predicting improved growth velocity and bone density during long-term recombinant human GH replacement. PMID- 8419614 TI - Hypothalamic-pituitary-gonadal function in prepubertal boys and girls with chronic renal failure. AB - Hypothalamic-pituitary-gonadal function was evaluated in 24 prepubertal children with chronic renal failure (CRF). Among the 17 boys, 5 were receiving conservative treatment and four long-term dialysis. Another eight boys were studied 6 months to 3.3 years after renal transplantation; their ages ranged from 5 years 8 months to 15 1/2 years. Among the girls, two patients were receiving conservative treatment and five long-term dialysis; their ages ranged from 3 1/2 years to 11 years 2 months. In boys with CRF, but not in those after transplantation, mean serum follicle-stimulating hormone 60 minutes after administration of gonadotropin releasing hormone (GnRH) was lower than in 18 control prepubertal boys (mean +/- SD: 2.53 +/- 1.34 vs 6.25 +/- 2.84 IU/L, respectively; p < 0.01). Testosterone steroidogenic capacity after 1 week of stimulation with human chorionic gonadotropin and androgen sensitivity (percentage of decrease of serum sex hormone-binding globulin 1 week after intramuscular administration of testosterone enanthate) were normal. In girls, no difference between those with CRF and a control group of 19 girls was found after intravenous administration of GnRH. However, after intramuscular administration of GnRH agonist, serum follicle-stimulating hormone concentration was lower in girls with CRF than in control girls (p < 0.02); six of seven control girls had an increase of serum estradiol to more than 55 pmol/L, whereas three of seven girls with CRF had no response, and serum follicle-stimulating hormone failed to increase after GnRH agonist therapy in two of these patients. We conclude that hypothalamic-pituitary function is not normal in some prepubertal boys and girls with CRF, particularly in those with low serum albumin concentrations. On the other hand, testicular and ovarian steroidogenic capacity is not impaired, and the biologic response to androgens in boys is preserved. PMID- 8419615 TI - Urine-to-blood carbon dioxide tension gradient and maximal depression of urinary pH to distinguish rate-dependent from classic distal renal tubular acidosis in children. AB - We determined the prevalence and clinical features of rate-dependent distal renal tubular acidosis (dRTA) in 31 children examined for possible renal tubular acidosis by measuring the urinary-minus-blood partial pressure of carbon dioxide (U-B PCO2) gradient, minimal urinary pH, and fractional excretion of bicarbonate. Of 20 patients with low U-B PCO2 gradients, nine could not lower urinary pH < or = 5.5, indicating classic dRTA, whereas 11 could lower urinary pH < or = 5.5, as described in rate-dependent dRTA. When patients with rate-dependent dRTA and classic (type I) dRTA were compared, there was no difference in the mean U-B PCO2 gradient or in clinical findings, including age, reason for referral, presence of nephrocalcinosis, or depression of linear growth. We conclude that children with rate-dependent dRTA are susceptible to at least some of the same sequelae as children with classic dRTA. Measurement of minimal urinary pH will not detect this subtle form of dRTA. Determination of the U-B PCO2 gradient should be considered a routine part of evaluation for suspected renal tubular acidosis in a child. PMID- 8419616 TI - Early manifestations of the carbohydrate-deficient glycoprotein syndrome. AB - We diagnosed the carbohydrate-deficient glycoprotein syndrome in five children who were seen during their first year of life with failure to thrive, feeding difficulties, psychomotor retardation, hypotonia, esotropia, inverted nipples, lipodystrophy, pericardial effusion, and hepatic dysfunction. Steatosis was observed in liver biopsy specimens, and cerebellar hypoplasia was present on computed tomography. The disorder is characterized by a complex carbohydrate deficiency in certain glycoproteins, notably transferrin, which can be used as a marker of the disease. The carbohydrate-deficient glycoprotein syndrome may be an important and easily identifiable cause of failure to thrive and neurologic dysfunction in infancy. The presence of the disorder in siblings of different gender and the finding of biochemical abnormalities in some unaffected parents suggest an autosomal recessive inheritance. PMID- 8419617 TI - Treatment of port-wine stains (capillary malformation) with the flashlamp-pumped pulsed dye laser. AB - Forty-three children with 49 separate port-wine stain vascular malformations were treated with the flashlamp-pumped pulsed dye laser at 585 nm. The patients ranged in age from 2 weeks to 14 years. Overall, 16% of patients had more than 95% resolution of their port-wine stains after an average of 4.8 (1 to 11) treatments. There was an average improvement of 69% in those lesions not clearing completely (average of 3.7 treatments). Lesions in patients less than 4 years of age were almost twice as likely to clear than were those in older children (20% vs 12%), and in fewer treatments (3.8 vs 6.5). An approximate 50% clearing was obtained with one treatment. Subsequent treatments resulted in an additional 10% clearing. There were no episodes of scarring or persistent pigmentary changes in any of the patients. Lesions on the hand and arm responded less well than lesions on the face, neck, and torso. The flashlamp-pumped pulsed dye laser has proved to be a safe and effective treatment modality for port-wine stain capillary malformations in infants and children. PMID- 8419618 TI - Acute fevers of unknown origin in young children in the tropics. AB - Bacteremia was documented in 14 of 156 previously healthy children with temperatures of at least 40.0 degrees C but without focal signs, seen in an emergency department; 116 children had malaria, and no infections were identified in 26. Concurrent malaria infection was frequent in children with bacteremia. The highly febrile child without focal signs in the tropics requires evaluation for bacteremia even when a diagnosis of malaria has been confirmed. PMID- 8419619 TI - Immunoglobulin allotypic markers in Kawasaki disease. AB - We investigated the possible relationship of the distribution of immunoglobulin allotypic markers for susceptibility to Kawasaki disease in Japanese, Japanese American, and white American populations. The kappa-chain allotype Km1 was present in 25.6% of sera from white patients with Kawasaki disease and in 14.4% of control sera (p < 0.01), and the combination of Km1 with Gm heterozygosity was present in 17.9% of white patients with Kawasaki disease and in 6.4% of control sera (p < 0.0001). In all populations studied, differences were observed between the patients with Kawasaki disease and the race-matched control subjects. The findings support the hypothesis that one or more unknown infectious agents may trigger genetically influenced immune responses that result in clinically recognizable Kawasaki disease. PMID- 8419620 TI - Reliability of a pulse oximeter in the detection of hyperoxemia. AB - To evaluate the reliability of the Nellcor N200 pulse oximeter in the detection of hyperoxemia (oxygen tension > 80 mm Hg), we obtained 213 simultaneous recordings and measurements of transcutaneous oxygen saturation (SO2) and arterial oxygen tension (PaO2) in 50 patients. During 95% of measurements with PaO2 > 80 mm Hg, SO2 was > or = 96%; SO2 was also > or = 96% in 20% of measurements with PaO2 < or = 80 mm Hg (false-positive results for hyperoxemia). With the upper alarm limit set at 95%, the pulse oximeter identified 95% of hyperoxemic instances while allowing PaO2 to be kept to > 60 mm Hg. PMID- 8419621 TI - Dietary protein intolerance in infants with transient methemoglobinemia and diarrhea. AB - Of 17 infants requiring hospitalization for primary soy or cow milk protein intolerance, six infants (35%) had transient methemoglobinemia. Reexposure to the offending protein caused diarrhea, metabolic acidosis, and transient methemoglobinemia in all patients. These six patients represented 65% of the total hospitalizations resulting from methemoglobinemia of any cause. PMID- 8419622 TI - Gastrointestinal hemorrhage associated with gastric polyps in Menkes disease. AB - We describe two infants with Menkes disease who had serious gastrointestinal bleeding from solitary gastric polyps. Hemorrhage in one patient was acute and proved fatal. Histopathologic examinations showed submucosal vascular ectasia with mucosal hyperplasia, edema, and ulceration. Gastric polyps may represent an underappreciated clinical abnormality in Menkes disease. PMID- 8419624 TI - Knee-chest position as a sign of increased intracranial pressure in children. AB - In three young children a newly acquired clinical sign--the knee-chest position- was associated with increased intracranial pressure. This sign may serve as an additional marker to suggest increased intracranial pressure in young children. PMID- 8419623 TI - Prospective management of a child with neonatal citrullinemia. AB - A patient with neonatal citrullinemia caused by severe deficiency of argininosuccinate synthetase was treated prospectively according to the currently accepted protocol. We gradually reduced the doses and then discontinued treatment with sodium benzoate and phenylacetate; blood glutamine levels were maintained in the normal range, but ammonia was mildly elevated. Growth and development progressed normally through 31 months of age. Some patients with citrullinemia can be successfully managed without daily sodium benzoate and phenylacetate therapy. PMID- 8419625 TI - Hereditary symphalangism with associated tarsal synostosis and hypophalangism. AB - Symphalangism is a rare genetic condition that may represent the earliest documentation of mendelian inheritance in man. The disorder results in interphalangeal joint fusion in the hands and feet. The authors review this rare condition and present a case study consisting of four generations with 15 affected family members. The association of multiple tarsal synostosis and the previously unreported associated occurrence of pedal hypophalangism in this pedigree is presented. PMID- 8419626 TI - Calcaneonavicular bar resection. A retrospective review. AB - The authors present a retrospective study evaluating the results of calcaneonavicular bar resection with interposition of the extensor digitorum brevis in ten patients with painful calcaneonavicular coalition. All operative procedures were supervised by one of the authors to ensure consistency in preoperative evaluation, surgical technique, and postoperative care. Results obtained from questionnaires showed significant improvement of preoperative symptoms with increased activity levels noted in all patients. PMID- 8419627 TI - Anatomical basis for congenital deformities of the lower extremities. Part I. The hip and thigh. AB - Congenital deformities frequently produce problems not always discernible at birth. Often, a period of time is required for the development of signs and symptoms. The present discussion presents the intrauterine anatomy of a midterm fetus relative to conditions of the hip and thigh. Cryomicrotomy is used in this study to present the best anatomical evidence of the morphology involved. PMID- 8419628 TI - Stress analysis of a flexible one-piece type first metatarsophalangeal joint implant. AB - Two-dimensional finite element analysis was used to obtain the magnitude and distribution of the stresses in models of two different designs of a flexible one piece double-stemmed first metatarsophalangeal joint implant. The loading and constraint conditions have been reported by previous researchers to occur in vivo at the joint during the push-off phase of the normal walking cycle. Based on the results obtained, the authors recommend the direction that future studies in this area should take. PMID- 8419629 TI - Heel spur surgery. A transverse plantar approach. PMID- 8419630 TI - Purpura fulminans secondary to pneumococcal sepsis in an asplenic patient. PMID- 8419631 TI - Particle analysis from small incision surgery. PMID- 8419632 TI - Histological re-evaluation of 101 intraoral salivary gland tumors by an EORTC study group. AB - Tumors of the salivary glands constitute a heterogeneous group of lesions of great morphologic variation and for this reason present many difficulties in histologic classification. The histologic slides of 101 consecutive intraoral salivary gland tumors of the Department of Oral Pathology of the Free University in Amsterdam were reviewed, retrospectively, by an EORTC-study group on salivary gland tumors. Complete concurrence of diagnosis was reached in 54 cases. In 33 cases there were minor disagreements, mostly related to subclassification. Major disagreements, relating to benign versus malignant, occurred in eight cases (7.9 per cent). PMID- 8419633 TI - Liposarcoma of the cheek: report of a case. AB - Liposarcomas of the head and neck region are rare. Those originating in the buccal mucosa cause special diagnostic and therapeutic difficulties. In this report, a predominantly well-differentiated liposarcoma of the cheek in a 32-yr old man is reported. The tumor continued to grow slowly over a period of 3 yr before definitive diagnosis was established. Radical maxillectomy was performed with total excision of the tumor. Recommendations for earlier and correct diagnosis and treatment of this rare neoplasm are discussed. PMID- 8419634 TI - Oral lesions in children born to HIV-1 positive women. AB - A cohort of 69 children born to HIV-1 positive women was studied to evaluate types, prevalences and relationships to clinical stages of HIV-1-related oral lesions. In addition, relationships among C. albicans biotypes, clinical features of oral candidiasis and HIV-1 disease were investigated. C. albicans biotypes did not correlate with clinical features of oral lesions, disease stages and CD4+ lymphocyte count. Of 8 patients with recurrent oral candidiasis, 4 changed clinical features and 5 changed biotype. Our study pointed out the high frequency (28.9%) of oral lesions, especially caused by fungi and the importance of the examination of the oral cavity in children born to HIV-1 positive women. PMID- 8419635 TI - Distinguishing features of an infectious molecular clone of the highly divergent and noncytopathic human immunodeficiency virus type 2 UC1 strain. AB - A full-length infectious molecular clone was derived from the noncytopathic human immunodeficiency virus type 2 UC1 strain (HIV-2UC1) that was originally recoverd from an individual from the Ivory Coast. Like the parental isolate, the molecularly cloned virus (HIV-2UC1mc or UC1 mc) demonstrates a reduced ability to induce syncytium formation, to kill cells, and to down-modulate the cell surface CD4 receptor in infected cells. Phylogenetic analysis of the DNA sequence of UC1mc revealed that it is the first full-length infectious molecular clone in the second HIV-2 subgroup previously identified by partial sequence analysis of the HIV-2D205 and HIV-2GH-2 strains. These highly divergent HIV-2 strains appear to be genetically equidistant from other HIV-2 and simian immunodeficiency virus SIVmac/sm strains. UC1mc is unlike any other HIV-2 or SIVmac/sm strain in that it lacks a cysteine residue at the proposed signal peptide cleavage site in Env. However, site-directed mutagenesis experiments indicate that this missing cysteine is not alone important in the noncytopathic phenotype of UC1mc. Like other HIV-2 and SIV strains, the UC1mc Env transmembrane protein (gp43) is mutated to a truncated form (gp34) after passage in certain T-cell lines. The UC1 molecular clone should be helpful in determining the genetic sequences associated with HIV-2 cytopathicity. PMID- 8419636 TI - Complete nucleotide sequence of a highly divergent human T-cell leukemia (lymphotropic) virus type I (HTLV-I) variant from melanesia: genetic and phylogenetic relationship to HTLV-I strains from other geographical regions. AB - The high prevalences of antibodies against human T-cell leukemia (lymphotropic) virus type I (HTLV-I) reported for remote populations in Papua New Guinea and the Solomon Islands and for some aboriginal populations in Australia have been verified by virus isolation. Limited genetic analysis of the transmembrane portion (gp21) of the envelope gene of these viruses indicates the existence of highly divergent HTLV-I strains in Melanesia. Here, we report the complete nucleotide sequence of an HTLV-I isolate (designated HTLV-IMEL5) from the Solomon Islands. The overall nucleotide divergence of HTLV-IMEL5 from the prototype HTLV IATK was approximately 8.5%. The degree of variability in the amino acid sequences of structural genes ranged between 3 and 11% and was higher (8.5 to 25%) for the regulatory (tax and rex) genes and the other genes encoded by the pX region. Since HTLV-IMEL5 was as distantly related to HTLV-II as to the other known HTLV-I strains, it could not have arisen from a reocmbinational event involving HTLV-II but rather might be an example of independent viral evolution in this remote population. These data provide important insights and raise new questions about the origin and global dissemination of HTLV-I. PMID- 8419637 TI - NS2 is required for efficient translation of viral mRNA in minute virus of mice infected murine cells. AB - Detailed analysis of five NS2 mutants of the autonomous parvovirus minute virus of mice (MVMp) has revealed the following. At low multiplicities of infection, NS2 mutants killed NB324K cells as well as wild-type (wt) MVM did and grew to high titers, while in contrast they grew poorly and did not readily kill murine A9 cells. Following CaPO4 transfection of murine fibroblasts, NS2 mutant infectious clones generated approximately 10-fold less monomer replicative-form DNA than wt and no detectable progeny single-stranded DNA. On nonmurine semipermissive NB324K cells, however, these mutant plasmid clones generated near wt levels of all replicative DNA forms. After infection of highly synchronized murine fibroblasts by NS2 mutant virus at inputs equivalent to those of the wt, mutant monomer replicative-form DNA was decreased 5- to 10-fold compared with that of the wt, and progeny single-stranded DNA accumulation was decreased to an even greater extent. Both total and cytoplasmic NS2 mutant RNA was decreased, but the amount of total viral mRNA generated, relative to accumulated viral DNA in the same experiments, was similar to that seen in wt infection. The accumulation of virus-generated proteins was also decreased in NS2 mutant infection; however, the magnitude of this decrease, compared with that of wt infections, was significantly greater than the concomitant decrease in mutant-generated levels of accumulated cytoplasmic RNA, and this effect was most dramatic for VP2. There was no such disparity between the relative accumulation of mutant-generated RNA and protein in cells permissive for the growth of these mutants. These results suggest that translation of MVM viral RNA is specifically reduced in NS2 mutant infection of restrictive cells. Because the affected viral proteins are required for the efficient production of viral replicative DNA forms, these results reveal a fundamental, although perhaps not the only, role for NS2 in parvovirus infection. PMID- 8419638 TI - The cytolytic activity of pulmonary CD8+ lymphocytes, induced by infection with a vaccinia virus recombinant expressing the M2 protein of respiratory syncytial virus (RSV), correlates with resistance to RSV infection in mice. AB - Previous studies demonstrated that the pulmonary resistance to respiratory syncytial virus (RSV) challenge induced by immunization with a recombinant vaccinia virus expressing the M2 protein of RSV (vac-M2) was significantly greater 9 days after immunization than at 28 days and was mediated predominantly by CD8+ T cells. In this study, we have extended these findings and sought to determine whether the level of CD8+ cytotoxic T-lymphocyte (CTL) activity measured in vitro correlates with the resistance to RSV challenge in vivo. Three lines of evidence documented an association between the presence of pulmonary CTL activity and resistance to RSV challenge. First, vac-M2 immunization induced pulmonary CD8+ CTL activity and pulmonary resistance to RSV infection in BALB/c (H-2d) mice, whereas significant levels of pulmonary CTL activity and resistance to RSV infection were not seen in BALB.K (H-2k) or BALB.B (H-2b) mice. Second, pulmonary CD8+ CTL activity was not induced by infection with other vaccinia virus-RSV recombinants that did not induce resistance to RSV challenge. Third, the peak of pulmonary CTL activity correlated with the peak of resistance to RSV replication (day 6), with little resistance being observed 45 days after immunization. An accelerated clearance of virus was not observed when mice were challenged with RSV 45 days after immunization with vac-M2. The results indicate that resistance to RSV induced by immunization with vac-M2 is mainly mediated by primary pulmonary CTLs and that this resistance decreases to very low levels within 2 months following immunization. The implications for inclusion of CTL epitopes into RSV vaccines are discussed in the context of these observations. PMID- 8419639 TI - Role of tumor necrosis factor alpha in activation and replication of the tat defective human immunodeficiency virus type 1. AB - Transcription of human immunodeficiency virus type 1 (HIV-1) depends on the function of the virus-encoded regulatory protein Tat, which interacts with the specific Tat response (TAR) element present in the leader sequence of all HIV-1 RNAs. In this study, we examined whether tumor necrosis factor alpha (TNF-alpha) can replace the requirement for a functional Tat protein. We found that TNF-alpha can induce expression of a latent, tat-defective virus and support its replication both in T cells and in primary mononuclear cells. Analysis of the transcriptional rate of the tat-defective HIV-1 transcriptional unit indicates that TNF-alpha stimulates the initiation of transcription but, in contrast to Tat protein, does not significantly reduce transcriptional polarity. Interestingly, we found that the processing of viral precursor proteins is altered in the absence of Tat. We propose that TNF-alpha-mediated induction of HIV-1 plays an essential role in the early stages of the virus life cycle and in viral latency. PMID- 8419640 TI - TAR RNA binding properties and relative transactivation activities of human immunodeficiency virus type 1 and 2 Tat proteins. AB - Using gel shift assays, we found that the human immunodeficiency virus type 1 (HIV-1) Tat protein (Tat-1) bound both HIV-1 and HIV-2 TAR RNAs with similar high affinities. In contrast, the HIV-2 Tat protein (Tat-2) bound only TAR-2 RNA with high affinity. We conclude that the weak in vivo activity of Tat-2 on the HIV-1 long terminal repeat that has been observed previously is likely the result of low affinity for TAR-1 RNA. Additionally, TAR-2 RNA was found to contain multiple specific binding sites for Tat proteins. GAL4-Tat fusion proteins were analyzed to compare the relative transactivation activities of Tat-1 and Tat-2 in the absence of requirements for binding to TAR RNAs. The GAL4-Tat-2 protein was found to transactivate synthetic promoters containing GAL4 binding sites at levels severalfold higher than did the GAL4-Tat-1 protein. PMID- 8419641 TI - Sequence variation of human endogenous retrovirus ERV9-related elements in an env region corresponding to an immunosuppressive peptide: transcription in normal and neoplastic cells. AB - Evolutionarily conserved sequences corresponding to an immunosuppressive region in retroviral transmembrane proteins were amplified by the polymerase chain reaction from human genomic DNA and reverse-transcribed RNA from one glioma, three pieces of macroscopically normal brain tissue, kidney, lymphocytes, cultured embryonic lung cells, and a rhabdomyosarcoma cell line. Amplification products (125 bp) from DNA and RNA from the glioma and RNA from one normal piece of brain tissue were cloned and sequenced (45 clones). A variety of sequences similar to ERV9 (75 to 93%) were identified. Amplification products were immobilized on nylon filters and hybridized to four different synthetic oligonucleotides derived from the sequenced clones. Sequences without the stop codon seen in ERV9 in this region, possibly encoding functional immunosuppressive proteins, were present in RNA amplificates from all samples. The various cell types showed different hybridization patterns with the four probes. The open reading frame sequences were identified in genomic Southern blots, one probe detecting about 10 copies and another detecting a single copy. Northern (RNA) blots of mRNA from various normal human tissues revealed 2.5-kb (e.g., lung) and 10-kb (e.g., placenta) transcripts hybridizing to one of the probes. PMID- 8419642 TI - Human immunodeficiency virus type 1 DNA integration: fine structure target analysis using synthetic oligonucleotides. AB - The target specificity of DNA strand transfer mediated by human immunodeficiency virus type 1 integrase was examined in vitro with synthetic oligonucleotides. Although insertion occurred at most locations in the target, some sites were preferred over others by at least 15-fold. Changing the nucleotide sequence of the target changed the distribution of preferred sites in complex ways, some of which included changes in target preference distant from the sequence alteration. Alignment of target sequences revealed that adenosine is preferred adjacent to the insertion site. Strand transfer occurred to within 2 nucleotides of the 3' end and to within 3 nucleotides of the 5' end of the target. This suggests that only 2 or 3 nucleotides flanking the target site are required for integration; such restricted contact with target DNA would allow integrase to insert the two ends of viral DNA into two closely spaced sites in host DNA, consistent with the concerted in vivo integration reaction that generates a 5-bp target duplication. PMID- 8419643 TI - A unique mitigator sequence determines the species specificity of the major late promoter in adenovirus type 12 DNA. AB - Human adenovirus type 12 (Ad12) cannot replicate in hamster cells, whereas human cells are permissive for Ad12. Ad12 DNA replication and late-gene and virus associated RNA expression are blocked in hamster cells. Early Ad12 genes are transcribed, and the viral DNA can be integrated into the host genome. Ad12 DNA replication and late-gene transcription can be complemented in hamster cells by E1 functions of Ad2 or Ad5, for which hamster cells are fully permissive (for a review, see W. Doerfler, Adv. Virus Res. 39:89-128, 1991). We have previously demonstrated that a 33-nucleotide mitigator sequence, which is located in the downstream region of the major late promoter (MLP) of Ad12 DNA, is responsible for the inactivity of the Ad12 MLP in hamster cells (C. Zock and W. Doerfler, EMBO J. 9:1615-1623, 1990). A similar negative regulator has not been found in the MLP of Ad2 DNA. We have now studied the mechanism of action of this mitigator element. The results of nuclear run-on experiments document the absence of MLP transcripts in the nuclei of Ad12-infected BHK21 hamster cells. Surprisingly, the mitigator element cannot elicit its function in in vitro transcription experiments with nuclear extracts from both hamster BHK21 and human HeLa cells. Intact nuclear topology and/or tightly bound nuclear elements that cannot be eluted in nuclear extracts are somehow required for recognition of the Ad12 mitigator. Electrophoretic mobility shift assays have not revealed significant differences in the binding of proteins from human HeLa or hamster BHK21 cells to the mitigator sequence in the MLP of Ad12 DNA or to the corresponding sequence in Ad2 DNA. We have converted the sequence of the mitigator in the MLP of Ad12 DNA to the equivalent sequence in the MLP of Ad2 DNA by site-directed mutagenesis. This construct was not active in hamster cells. When the Ad12 mitigator, on the other hand, was inserted into the Ad2 MLP, the latter's function in hamster cells was not compromised. Deletions in the 5' upstream region of the Ad12 MLP have provided evidence for the existence of additional sequences that codetermine the deficiency of the Ad12 MLP in hamster cells. The amphifunctional YY1 protein from HeLa cells can bind specifically to the mitigator and to upstream elements of the MLP of Ad12 DNA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419644 TI - Human immunodeficiency viruses containing heterologous enhancer/promoters are replication competent and exhibit different lymphocyte tropisms. AB - The human immunodeficiency virus (HIV) type 1 long terminal repeat (LTR) contains binding sites for nuclear factor kappa B (NF-kappa B) and the constitutively expressed transcription factor Sp1, both of which are highly conserved in HIV and simian immunodeficiency virus isolates. To delineate the effects of these motifs on the replicative capacity of HIV and to explore the possibility of extending the virus host range, known heterologous enhancer/promoters were inserted into the HIV-1 LTR in place of the NF-kappa B and Sp1 binding sites. The effects of these substitutions on viral replication in transfected HeLa cells and on HIV infection of human peripheral blood lymphocytes or continuous T-leukemia cell lines were evaluated. HIVs in which the NF-kappa B/Sp1 enhancer plus the downstream TATA element were replaced with heterologous enhancer/promoters were also constructed. Viruses containing the human cytomegalovirus immediate-early enhancer exhibited infectious kinetics similar to that of wild-type HIV in activated human peripheral blood lymphocytes and AA2 cells but replicated less efficiently in H9 and CEM cells. These studies indicate that heterologous enhancer elements are capable of restoring Tat responsiveness to the HIV LTR in the context of directing reporter gene expression as well as in the production of infectious progeny virions. PMID- 8419645 TI - Biologic importance of neuraminidase stalk length in influenza A virus. AB - To investigate the biologic importance of the neuraminidase (NA) stalk of influenza A virus, we generated mutant viruses of A/WSN/33 (H1N1) with stalks of various lengths (0 to 52 amino acids), by using the recently developed reverse genetics system. These mutant viruses, including one that lacked the entire stalk, replicated in tissue culture to the level of the parent virus, whose NA stalk contains 24 amino acid residues. In eggs, however, the length of the stalk was correlated with the efficiency of virus replication: the longer the stalk, the better the replication. This finding indicates that the length of the NA stalk affects the host range of influenza A viruses. The NA stalkless mutant was highly attenuated in mice; none of the animals died even after intranasal inoculation of 10(6) PFU of the virus (the dose of the parent virus required to kill 50% of mice was 10(2.5) PFU). Moreover, the stalkless mutant replicated only in the respiratory organs, whereas the parent virus caused systemic infection in mice. Thus, attenuation of the virus with the deletion of the entire NA stalk raises the possibility of its use as live vaccines. PMID- 8419646 TI - Cell proteins bind to sites within the 3' noncoding region and the positive strand leader sequence of measles virus RNA. AB - The genomic 3' noncoding region (NCR) of nonsegmented negative-strand RNA viruses contains recognition site(s) for the polymerase complex, while the RNA plus strand leader sequence (LS) is probably involved in RNA encapsidation. It is known that host-encoded factors play a role in transcription and replication of some of this group of viruses. Here we report that cellular proteins interact with the genomic 3' NCR and with the plus-strand LS RNA of an important human pathogen, measles virus (MV), a member of the family Paramyxoviridae. Using gel retardation assay and RNA footprinting analysis, we demonstrated that in Vero cells, host-encoded proteins bind specifically to domains within these two sequences. A polypeptide of about 20 kDa binding to the 3' NCR and two polypeptides of about 22 and 30 kDa interacting with plus-strand LS were detected by RNA-protein UV cross-linking. Different RNA-binding activities were found in cells differing in permissiveness to MV replication. The results suggest a role for host-encoded proteins in MV replication. PMID- 8419647 TI - The Tacaribe arenavirus small zinc finger protein is required for both mRNA synthesis and genome replication. AB - An antiserum to a peptide of the Tacaribe virus Z protein was used to determine whether this small Zn(2+)-binding protein was required for viral RNA synthesis in infected cell extracts. Specific immunodepletion of the extracts invariably reduced genome synthesis to near background levels, but strong effects on mRNA synthesis occurred only early in the infection or when mRNA synthesis was relatively weak. Our results suggest that the Z protein is required for both mRNA and genome synthesis, but in somewhat different manners. PMID- 8419648 TI - Analysis of multiple mRNAs from pathogenic equine infectious anemia virus (EIAV) in an acutely infected horse reveals a novel protein, Ttm, derived from the carboxy terminus of the EIAV transmembrane protein. AB - Transcription of pathogenic equine infectious anemia virus (EIAV) in an acutely infected horse was examined by using the polymerase chain reaction and nucleotide sequencing. Four spliced transcripts were identified in liver tissue, in contrast to the multiplicity of alternatively spliced messages reported for in vitro propagated human immunodeficiency virus, simian immunodeficiency virus, and, to a lesser extent, EIAV. Nucleotide sequence analysis demonstrated that three of these mRNAs encode known viral proteins: the envelope precursor, the product of the S2 open reading frame, and the regulatory proteins Tat and Rev. The fourth transcript encodes a novel Tat-TM fusion protein, Ttm. Ttm is a 27-kDa protein translated from the putative tat CTG initiation codon and containing the carboxy terminal portion of TM immediately downstream from the membrane-spanning domain. p27ttm is expressed in EIAV-infected canine cells and was recognized by peptide antisera against both Tat and TM. Cells transfected with ttm cDNA also expressed p27ttm, which appeared to be localized to the endoplasmic reticulum or Golgi apparatus by indirect immunofluorescence. The carboxy terminus of lentiviral TM proteins has previously been shown to influence viral infectivity, growth kinetics, and cytopathology, suggesting that Ttm plays an important role in the EIAV life cycle. PMID- 8419649 TI - CD4 molecules with a diversity of mutations encompassing the CDR3 region efficiently support human immunodeficiency virus type 1 envelope glycoprotein mediated cell fusion. AB - The third complementarity-determining region (CDR3) within domain 1 of the human CD4 molecule has been suggested to play a critical role in membrane fusion mediated by the interaction of CD4 with the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein. To analyze in detail the role of CDR3 and adjacent regions in the fusion process, we used cassette mutagenesis to construct a panel of 30 site-directed mutations between residues 79 and 96 of the full-length CD4 molecule. The mutant proteins were transiently expressed by using recombinant vaccinia virus vectors and were analyzed for cell surface expression, recombinant gp120-binding activity, and overall structural integrity as assessed by reactivity with a battery of anti-CD4 monoclonal antibodies. Cells expressing the CD4 mutants were assayed for their ability to form syncytia when mixed with cells expressing the HIV-1 envelope glycoprotein. Surprisingly in view of published data from others, most of the mutations had little effect on syncytium-forming activity. Normal fusion was observed in 21 mutants, including substitution of human residues 85 to 95 with the corresponding sequences from either chimpanzee, rhesus, or mouse CD4; a panel of Ser-Arg double insertions after each residue from 86 to 91; and a number of other charge, hydrophobic, and proline substitutions and insertions within this region. The nine mutants that showed impaired fusion all displayed defective gp120 binding and disruption of overall structural integrity. In further contrast with results of other workers, we observed that transformant human cell lines expressing native chimpanzee or rhesus CD4 efficiently formed syncytia when mixed with cells expressing the HIV-1 envelope glycoprotein. These data refute the conclusion that certain mutations in the CDR3 region of CD4 abolish cell fusion activity, and they suggest that a wide variety of sequences can be functionally tolerated in this region, including those from highly divergent mammalian species. Syncytium formation mediated by several of the CDR3 mutants was partially or completely resistant to inhibition by the CDR3-directed monoclonal antibody L71, suggesting that the corresponding epitope is not directly involved in the fusion process.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419650 TI - Complex-type N-linked oligosaccharides of gp120 from human immunodeficiency virus type 1 contain sulfated N-acetylglucosamine. AB - The major envelope glycoproteins gp120 and gp41 of human immunodeficiency virus type 1, the causative agent for human AIDS, contain numerous N-linked oligosaccharides. We report here our discovery that N-acetylglucosamine residues within the complex-type N-linked oligosaccharides of both gp120 and its precursor, gp160, are sulfated. When human Molt-3 cells persistently infected with human T-cell leukemia virus IIIB were metabolically radiolabeled with 35SO4, gp160, gp120, and to some extent gp41 were radiolabeled. The 35SO4-labeled oligosaccharides were quantitatively released by N-glycanase treatment and were bound by immobilized Ricinus communis agglutinin I, a lectin that binds to terminal beta-galactosyl residues. The kinetics of release of sulfate upon acid hydrolysis from 35SO4-labeled gp120 indicate that sulfation occurs in a primary sulfate ester linkage. Methylation analysis of total glycopeptides from Molt-3 cells metabolically radiolabeled with [3H]glucosamine demonstrates that sulfation occurs at the C-6 position of N-acetylglucosamine. Fragmentation of the gp120 derived 35SO4-labeled glycopeptides by treatment with hydrazine and nitrous acid and subsequent reduction generated galactosyl-anhydromannitol-6-35SO4, which is the expected reaction product from GlcNAc-6-sulfate within a sulfated lactosamine moiety. Charge analysis of the [3H]galactose- and [3H]glucosamine-labeled glycopeptides from gp120 and gp160 indicates that approximately 14% of the complex-type N-linked oligosaccharides are sulfated. PMID- 8419652 TI - Unspecified reform tops agenda. PMID- 8419651 TI - Recombination and polymerase error facilitate restoration of infectivity in brome mosaic virus. AB - The tRNA-like structure present in the 3' noncoding region of each of the four virion RNAs of brome mosaic virus possesses a conserved A-67-U-A-65 (67AUA65) sequence. Four mutations in this region (67UAA65, 67GAA65, and 67CAA65, each with a double base change, and 67GUA65, containing a single point mutation), previously shown in vitro to be defective in minus-strand promoter function, were introduced into full-length genomic RNAs 2 and 3, and their replicative competence was analyzed in barley protoplasts. All four RNA 3 mutants were capable of replication, although progeny plus-sense RNA 3 accumulation was only 12 to 42% of that of the wild type. Replication of RNA 2 transcripts bearing these mutations was even more severely debilitated; the accumulation of each mutant progeny plus-strand RNA 2 was < 10% of that of the wild type. Analysis of mutant RNA 3 progeny recovered from local lesions induced in Chenopodium hybridum and systemic infections in barley (Hordeum vulgare) plants revealed that the mutant base at position 67 from the 3' end had in each case been modified to an A. These changes generated RNAs with functional pseudorevertant (67AAA65 for mutants 67UAA65, 67GAA65, and 67CAA65) or revertant (67GUA65-->67AUA65) sequences. In most instances, the presence of internal markers permitted discrimination between polymerase error and RNA recombination as the process by which sequence restoration occurred. The pseudorevertant sequence was found to be capable of persistence during subsequent propagation in plants when present on RNA 3 but not when present on RNA 2. These data document the fluidity of the RNA genome and reveal situations in which polymerase error or recombination can function preferentially to restore an optimal sequence. They also support the concept that RNA viruses frequently exist as quasispecies and have implications concerning evolutionary strategies for positive-strand RNA viruses. PMID- 8419653 TI - Two new reports help put nation's No. 1 killer disease challenges into perspective for 1993. PMID- 8419654 TI - Revascularization still the goal, strategy differs as cardiologists consider clinical trial results. PMID- 8419655 TI - Desert Storm veterans now may donate blood; others call for discussion of donor tests. PMID- 8419656 TI - From the Institute of Medicine. PMID- 8419657 TI - From the Centers for Disease Control and Prevention. Imported human rabies- France, 1992. PMID- 8419658 TI - From the Centers for Disease Control and Prevention. Availability of sulfadiazine -United States. PMID- 8419659 TI - Alcohol and the future of Native Americans. PMID- 8419660 TI - Single-source financing: pitching the curve ball. PMID- 8419662 TI - The causes of colon cancer. PMID- 8419661 TI - Tuberculosis in anergic HIV-infected drug users. PMID- 8419663 TI - Tick bite prevention: get naked? PMID- 8419664 TI - Antineoplastons: the controversy continues. PMID- 8419665 TI - Antineoplastons: the controversy continues. PMID- 8419666 TI - Smoking and risk of Graves' disease. AB - OBJECTIVE: To assess if smoking is associated with Graves' disease and, if so, to ascertain whether this association persists when controlling for confounding factors. DESIGN: Consecutive entry case-control study with two age- and sex matched control subjects from two different populations per case patient. SETTING: University hospital. PATIENTS: Five groups were studied: (1) Graves' ophthalmopathy and Graves' hyperthyroidism (n = 100; divided in four subgroups according to the severity of the eye disease); (2) Graves' hyperthyroidism without clinical eye involvement (n = 100); (3) sporadic nontoxic goiter (n = 100); (4) autoimmune hypothyroidism (n = 75); and (5) toxic nodular goiter (n = 75). The study comprised 200 subjects from a hospital-based population, and 200 from a population-based group served as control subjects. MAIN OUTCOME MEASURE: Smoking status was determined from a questionnaire at the time of onset of the disease to exclude any effect of the disease itself on smoking. RESULTS: Smoking greatly increased the risk for Graves' ophthalmopathy (odds ratio, 7.7; 95% confidence interval, 4.3 to 13.7), but patients with Graves' hyperthyroidism alone were also more often smokers than control subjects (odds ratio, 1.9; 95% confidence interval, 1.1 to 3.2). Smoking was not associated with the other thyroid diseases studied. Essentially similar results were obtained after adjustment for differences in education between case patients and control subjects. Among the patients with Graves' ophthalmopathy, smokers had more severe eye disease than nonsmokers, but no association was found between the number of cigarettes smoked per day or the duration of smoking and the severity of the ophthalmopathy. However, there was a significant increase in the odds ratios in patients with more severe eye disease. CONCLUSIONS: Smoking is associated with Graves' disease, and it especially increases the risk for the development of more severe ophthalmopathy. Thus, smoking appears to be one of the multiple factors inducing Graves' disease in genetically predisposed individuals. PMID- 8419667 TI - Body fat distribution and 5-year risk of death in older women. AB - OBJECTIVE: To test the hypothesis that both body mass index (expressed as the ratio of weight in kilograms per height in meters squared) and the ratio of waist circumference to hip circumference are positively associated with mortality risk in older women. DESIGN: Prospective cohort study with a 5-year follow-up period. SETTING: General community. PARTICIPANTS: Random sample of 41,837 Iowa women aged 55 to 69 years. MAIN OUTCOME MEASURE: Total mortality (1504 deaths). MAIN RESULTS: Body mass index, an index of relative weight, was associated with mortality in a J-shaped fashion: rates were elevated in the leanest as well as in the most obese women. In contrast, waist/hip circumference ratio was strongly and positively associated with mortality in a dose-response manner. Adjusted for age, body mass index, smoking, education level, marital status, estrogen use, and alcohol use, a 0.15-unit increase in waist/hip circumference ratio (eg, a 15-cm [6-in] increase in waist measurement in a woman with 100-cm [40-in] hips) was associated with a 60% greater relative risk of death. The observed associations were not explained to any great degree by bias from weight loss prior to baseline or higher early deaths among lean participants. CONCLUSIONS: Waist/hip circumference ratio is a better marker than body mass index of risk of death in older women. Waist/hip circumference ratio should be measured as part of routine surveillance and risk monitoring in medical practice. PMID- 8419668 TI - A prospective study of blood pressure and serum creatinine. Results from the 'Clue' Study and the ARIC Study. AB - OBJECTIVES: To describe associations of past and current blood pressure levels with serum creatinine levels and hypercreatinemia in the general population. POPULATION: 1399 middle-aged residents of Washington County, Maryland, who had their blood pressure measured during a cancer screening campaign in 1974 (the "Clue" Study) and had their blood pressure and serum creatinine level measured in an atherosclerosis risk factors study from 1986 through 1989 (the Atherosclerosis Risk in Communities [ARIC] Study). DESIGN: Nonconcurrent prospective study. The outcome variables were serum creatinine level and hypercreatinemia (serum creatinine > 115 mumol/L in men, > 97 mumol/L in women) measured from 1986 through 1989. Main predictors were 1986-1989 blood pressure values (cross sectional association) and 1974 blood pressure values (longitudinal association). Gender-adjusted associations were assessed and compared by linear and logistic regression. RESULTS: Both serum creatinine and hypercreatinemia were better predicted by past than by current blood pressure values. Creatinine values exhibited a gradual and statistically significant association with blood pressure levels measured in 1974, even across "normal" values of blood pressure and creatinine. The association with 1986-1989 blood pressure measurements was weaker and nonsignificant. The odds of hypercreatinemia in 1986-1989 were increased 1.5 fold to twofold, with a 20 mm Hg increment in 1974 blood pressure values, but the odds remained constant across 1986-1989 blood pressure values. CONCLUSIONS: Blood pressure and creatinine level are associated in the general population. The observed association was stronger when a number of years had elapsed between the assessments of blood pressure and creatinine level. These findings are consistent with the hypothesis that blood pressure elevations, even below the hypertensive range, may induce early renal damage. PMID- 8419669 TI - Achieving a uniform federal primary care policy. Opportunities presented by national health reform. PMID- 8419670 TI - An outbreak of mud-wrestling-induced pustular dermatitis in college students. Dermatitis palaestrae limosae. AB - OBJECTIVE: To investigate an outbreak of gram-negative folliculitis in relation to a common exposure, mud wrestling, and identify risk factors for dermatitis among those who mud wrestled. DESIGN: Case-control study. SETTING: University of Washington, Seattle. PARTICIPANTS: Two college-residence groups of students. RESULTS: Cultures from affected students and from mud similar to that used for wrestling yielded Enterobacteriaceae. The odds ratio associated with mud wrestling was 79.5 (95% confidence interval, 13.9 to 895.4). Increased time spent wrestling was associated with increased risk. Skin trauma during wrestling or immersion in the mud increased the risk of infection (odds ratio, 23.1; 95% confidence interval, 1.7 to 1440.4). CONCLUSIONS: Mud wrestling is one cause of pustular follicular dermatitis. Trauma to the skin may be a necessary cofactor for the development of infection. PMID- 8419671 TI - NIH Consensus conference. Triglyceride, high-density lipoprotein, and coronary heart disease. NIH Consensus Development Panel on Triglyceride, High-Density Lipoprotein, and Coronary Heart Disease. PMID- 8419672 TI - MinnesotaCare (HealthRight). Myths and miracles. PMID- 8419673 TI - Antibiotic therapy for otitis media with effusion. A response from Pittsburgh. PMID- 8419674 TI - Tobacco and Graves' disease. Smoking gun or smoke and mirrors? PMID- 8419675 TI - Examining motion in the cervical spine. II: Characterization of coupled joint motion using an opto-electronic device to track skin markers. AB - Analysis of coupled motion in the cervical spine may be useful in helping to identify injuries. In order to investigate this possibility, the nature of coupled motion in the spine and previous investigations on this subject are reviewed here. An enhanced set of displays are developed for an existing opto electronic device employed for the non-invasive measurement of movement in the upper spine. This instrument consists of a high resolution motion analysis system which tracks small infrared emitting diodes (IREDs). Kinematic data for the motion of the markers is processed and absolute coordinates for the location of each IRED at any time are tabulated; coupled motion with respect to a fixed calibration frame, as well as for vertebrae relative to each other, is deduced from these. Overall analysis provided by the original device includes assessment of cervical lordosis, thoracic kyphosis, and inter-segmental mobility. Characterization of coupled motion, in particular, involves a series of plots showing principal versus secondary motion. Principal movements include flexion extension, lateral bending, and axial rotation, corresponding to motion in the sagittal, transverse, and horizontal planes, respectively. Mobility is represented in terms of the direction angles made by virtual vectors orthogonal to the planes made by markers on the head, neck, and shoulders. Development of the enhanced displays and the required refinements are described. Precision of the deduced angles is found to be approximately 1 degree. This representation of coupled motion is expected to be valuable in improving the accuracy of attempts to identify normal versus pathological motion in the cervical spine. PMID- 8419676 TI - Role of the calcaneal heel pad and polymeric shock absorbers in attenuation of heel strike impact. AB - The capacity of the calcaneal heel pad, with and without augmentation by a polymeric shock absorbing material (Sorbothane 0050), to attenuate heel strike impulses has been studied using five fresh human cadaveric lower leg specimens. The specimens, instrumented with an accelerometer, were suspended and impacted with a hammer; a steel rod was similarly suspended and impacted. The calcaneal heel pad attenuated the peak accelerations by 80%. Attenuations of up to 93% were achieved by the shock absorbing material when tested against the steel rod; however, when tested in series with the calcaneal heel pad, the reduction in peak acceleration due to the shock absorbing material dropped to 18%. Any evaluation of the effectiveness of shock absorbing shoe materials must take into account their mechanical interaction with the body. PMID- 8419677 TI - Testing of small connective tissue specimens for the determination of the mechanical behaviour of atherosclerotic plaques. AB - The tearing of the cap of atheromatous plaques is the commonest cause of thrombosis in human coronary arteries. It has been proposed that tearing arises because of structural weakening of the cap's connective tissue around the tear. To test this hypothesis we compared the mechanical properties of the intact edges of torn plaque caps and unbroken caps. Owing to limitations in plaque size, a purpose-built tensometer was developed to study stress-strain relationships of the small connective tissue specimens. The design of the tensometer is reported and was shown to detect accurately, minor differences in connective tissues and to generate complete stress-curves with computer-assisted image analysis. PMID- 8419678 TI - Repetitive elastance measurement as a means of recording pharmacological responses in vascular smooth muscles. AB - We describe a method of recording pharmacological responses in isolated arterial segments, based on the direct measurement of vascular wall elastance. The segment outflow is intermittently occluded, and the elastance determined from the subsequent pressure-flow relationship. Since the measurement cycle can be repeated 12 times per minute, rapid pharmacological responses can be studied. The method is capable of demonstrating a sigmoid dose-response relationship to noradrenaline added to the perfusate. PMID- 8419679 TI - Image analysis in the evaluation of biomaterials. AB - An examination regime, based on a computer-aided image analysis system, has been developed for the quantitative evaluation of the local tissue response to biomaterials. This procedure involves the immunoenzymic staining of tissue sections using monoclonal antibodies specific for certain inflammatory cell types. An avidin-biotin-horseradish peroxidase staining method is used to identify antibody binding sites and the sections are assayed using a computer aided image analysis system. This regime facilitates the rapid and accurate measurement of 30 cell related parameters in sections stained for macrophages, polymorphonuclear leucocytes, and other cells. PMID- 8419680 TI - The forces acting on the human calcaneus. AB - The forces acting on the human calcaneus are analysed for a static standing posture. An optimization procedure is invoked to solve the statically indeterminate set of forces which could arise from muscles, ligaments and reactions between the calcaneus and other bones and also the ground. Two alternative objective functions are investigated; that which returns no active muscle force is considered the more realistic, since electromyographic studies show that in static standing only minor muscle activity occurs to counteract body sway. This set of forces is then applied to the bone for finite element stress analysis to produce stress trajectories, which are then validated against both the trabecular configuration of the cancellous bone of the calcaneus and also the distribution of the orientations of the hydroxyapatite crystal major and minor axes. PMID- 8419681 TI - Examining motion in the cervical spine. I: Imaging systems and measurement techniques. AB - Instruments for measuring mobility in the cervical spine range from plumb-lines and inclinometers to sophisticated optoelectronic systems. In order to investigate the need and possible uses for an enhancement to a new diagnostic instrument, we examine some of the available diagnostic systems suitable for cervical motion analysis. These should be of practical use in a clinical setting for the diagnosis of soft tissue injuries. We begin by evaluating the respective roles of plain radiographs, cineradiography, computer tomography, and magnetic resonance imaging in examining the cervical spine. Then we consider Moire photography, inclinometers, and some opto-electronic scanners, as well as the mathematical techniques needed to correlate skin and spine motion with these devices. We find that there does not appear to be an effective non-invasive tool for comprehensive clinical cervical motion analysis; in particular, coupled joint motion is inadequately quantified. Improperly diagnosed cervical spine injuries, such as hyperextension and hyperflexion, may result in chronic long-term effects. Therefore, instrumentation that would permit objective, routine clinical evaluation of patients could help to avoid such situations. PMID- 8419682 TI - Improving limb flexion in FES gait using the flexion withdrawal response for the spinal cord injured person. AB - In the restoration of gait for paraplegics using functional electrical stimulation, the method most commonly used to produce hip flexion for the swing phase of gait has been the elicitation of the flexion withdrawal response. Several problems have been noted with the response: there is a decrease in the magnitude of the hip flexion to repeated stimuli (habituation); long latency; and inhibition of the response when stimulated bilaterally. These have been characterized and methods for overcoming the problems tested. Results show that increasing stimulation frequency reduces latency. Habituation can be reduced in some subjects by multiplexing two sites of stimulation. Habituation can further be reduced by applying single high-intensity pulses and this has been used in a one-step-ahead controller for regulating hip flexion angle. Inhibition due to bilateral stimulation had been significantly reduced by altering the timing of the stimulation to the two legs. PMID- 8419683 TI - Digital dynamic range expansion applied to X-ray densitometric analysis of total hip replacement. AB - Due to the presence of the stiff prosthetic stem fitted in the medullary canal during total hip replacement, the surrounding cortex of the femur changes its density over time. This bone remodelling takes place with every type of total hip prosthesis; however, its intensity may vary between prostheses and patients. In the worst cases this process can lead to the late failure of the implant. To monitor such bone density evolution, we are developing a tailored Computer-aided Densitometric Image Analysis system (the major part of this our system uses an 8 bit commercial hardware with 256 levels of grey). The equivalent dynamic range of an X-ray picture is about 10 bits. In this paper we present a method to overcome these hardware limitations by improving the software. Using a double-exposure acquisition it is possible to build a 9-bit image that is good enough for most applications involving bone density measurement. PMID- 8419684 TI - An intermediate loading rate technique for the determination of mechanical properties of human femoral cortical bone. AB - An experimental method to evaluate the mechanical properties of human femoral cortical bone under an intermediate strain rate of 4 x 10(-2) s-1 is presented. The dynamic loading was developed by dropping weights from various heights; results were obtained for Young's modulus and Poisson's ratio when the specimens were loaded along the longitudinal axis of the femur. The results were compared with those under a slow strain rate of 2 x 10(-5) s-1 and a fast strain rate of 1 x 10(-2) s-1. It was found that Young's modulus for the intermediate strain rate is 10.5% higher than for the slow strain rate and 10% lower than for the fast strain rate. Poisson's ratio did not show any significant variation for the above three strain rates. The results were compared with those given previously by other investigators. PMID- 8419685 TI - A biomechanical impact test system for head and facial injury assessment and model development. AB - A biomechanical test system has been developed and validated to conduct controlled uniaxial impact experiments of head and facial trauma. The design reduces off-axis accelerations which are not in the direction of impact and allows accurate positioning of test specimens. Impact forces, displacement histories, impulses at impact and spectral responses are compared to free-fall test results at contact velocities representative of facial injuries (2.5, 3.1 and 3.8 m s-1). Models based on the experimental results are developed to reveal stiffness and inertial properties of impact for use in the design of biomechanically protective steering wheels, air bags and other potential impact structures. The results indicate that the system provides a flexible yet controllable method for positioning and testing impact structures reliably. PMID- 8419686 TI - Cardiovascular applications of the ALOPEX optimization technique. AB - ALOPEX is a general optimization process incorporating a cost function containing a large number of parameters which may be simultaneously adjusted until the cost function reaches an optimum (maximum or minimum); local extremes are avoided by introducing random noise into the procedure. In this paper, ALOPEX is incorporated into a simple haemodynamic study in which an electric analogue model of the left ventricle is used to develop equations of myocardial stroke work. Pilot experiments were undertaken in rabbits (n = 5) to gauge the effectiveness of this optimizing technique. In the control state, calculated stroke work for the rabbit was determined to be 50 +/- 7 mmHg ml, while ALOPEX predicted a stroke work of 51 +/- 7 mmHg ml. ALOPEX is capable of following changing cardiovascular states when pharmacological agents are introduced. For example, after nitroprusside treatment, stroke work was reduced by 38 +/- 6% (P < 0.05) while ALOPEX predicted a 42 +/- 4% reduction from baseline (P < 0.05). Methoxamine treatment increased stroke work by 74 +/- 34%, while ALOPEX predicted a 73 +/- 43% increase above control values. There were no statistical differences between calculated and ALOPEX predicted values. Individual model parameters such as maximum left ventricular elastance (Emax) and left ventricular end diastolic volume (EDV) were also predicted correctly by ALOPEX. We have found that the ALOPEX optimization technique is useful in predicting components of multi parametric functions. In particular, we have shown it to be adaptable to a simple haemodynamic model. PMID- 8419687 TI - An alternative to screws for plating osteoporotic bone. AB - There are several problems with existing techniques used to plate broken bones. One of the most serious is that of screws pulling out during insertion or healing in patients suffering from osteoporosis. This paper investigates the mechanism of this pulling out, and proposes an alternative device to attach plates to weak bones. PMID- 8419688 TI - Comment on: the energetics of paraplegic walking. PMID- 8419689 TI - Drijber and Finlay (1992)--universal joint slippage as a cause of Hofmann half frame external fixator failure. J Biomed Eng 1992; 14: 509-15. PMID- 8419690 TI - Functional foregut abnormalities in Barrett's esophagus. AB - The factors predisposing to the development of Barrett's esophagus in patients with gastroesophageal reflux disease are unclear. We compared symptoms, esophageal acid and alkaline exposure (pH < 2, < 3, < 4, and > 7), lower esophageal sphincter resistance, esophageal clearance function, the gastric secretory state, gastric emptying, and duodenogastric reflux in 15 patients with Barrett's esophagus with 24 patients with esophagitis and with 22 normal subjects. Compared with patients with esophagitis, patients with Barrett's esophagus had less heartburn and regurgitation but had an increased frequency and duration of reflux episodes and percent time pH less than 2, less than 3, less than 4, and pH greater than 7 on ambulatory 24-hour esophageal pH monitoring. This was associated with a decreased lower esophageal sphincter resistance, a decreased contraction amplitude in the distal area of the esophagus, an increased frequency of nonperistaltic contractions and contractions less than 30 mm Hg on 24-hour ambulatory esophageal motility monitoring, increased basal and stimulated gastric acid secretion, and a higher prevalence of excessive duodenogastric reflux. These data show that despite less symptoms patients with Barrett's esophagus have a markedly increased esophageal acid and alkaline exposure compared with patients with esophagitis. This appears to be because of persistent reflux of highly concentrated gastric acid and duodenal contents across a mechanically defective lower esophageal sphincter in combination with inefficient esophageal clearance function. PMID- 8419691 TI - Biventricular repair of hypoplastic right ventricle assisted by pulsatile bidirectional cavopulmonary anastomosis. AB - The right ventricle in patients with severe outflow obstruction or atresia and a small tricuspid valve often remains too hypoplastic even after optimal palliation to tolerate biventricular repair with closure of the atrial septal defect. In these patients, nonpulsatile cavopulmonary (Glenn) anastomosis has traditionally facilitated biventricular repair. In 1989, Billingsley and associates reported the addition of a bidirectional cavopulmonary anastomosis to the definitive biventricular repair in patients with hypoplastic right ventricle, pulmonary atresia, and intact ventricular septum. The atrial septal defect was left open with an adjustable snare for later closure. We report five patients with hypoplastic right ventricle (mean diastolic volume 48.4%, mean stroke volume 40.2% of predicted value) who had the atrial septal defect closed at the time of the biventricular repair. Four patients, who had the bidirectional cavopulmonary anastomosis supplementing the biventricular repair, had no evidence of excessive right atrial or superior vena cava hypertension postoperatively. One patient, who had atypical tetralogy of Fallot with tricuspid stenosis, developed recurrent pericardial tamponade and marked hepatomegaly following conventional tetralogy repair with closure of the atrial septal defect. These complications were controlled with the addition of bidirectional cavopulmonary anastomosis 2 months later. Postoperative hemodynamic or Doppler studies in these patients revealed pulsatile flow in the entire pulmonary artery system, including the artery distal to the Glenn anastomosis. This modification of biventricular repair allows primary closure of the atrial septal defect and provides pulsatile arterial flow in the entire pulmonary artery, even when the right ventricle is significantly hypoplastic. PMID- 8419692 TI - Left ventricular pressure overload during postnatal development. Effects on coronary vasodilator reserve and tolerance to hypothermic global ischemia. AB - The effect of pressure overload during postnatal development on (1) coronary reserve and (2) tolerance to cardioplegic arrest and hypothermic ischemia was investigated. Left ventricular pressure overload was induced in 1-week-old rats by aortic constriction (group AC), with control rats (group C) undergoing sham operation. Relative to group C rats, left ventricular weight/body weight ratio in group AC increased by 50% to 80% at 3 weeks of age and by 100% to 120% at 6 weeks of age. At these ages, hearts were isolated from group C and AC rats and Langendorff-perfused with bicarbonate buffer at perfusion pressures of 75 and 110 mm Hg, respectively. In the first study, minimal coronary vascular resistance was assessed during perfusion with adenosine (10 mumol/L). There was no difference in total minimal coronary vascular resistance between groups C and AC at 3 weeks of age. Total minimal coronary vascular resistance decreased with increasing cardiac mass between 3 and 6 weeks of age in group C (from 6.9 +/- 0.4 to 4.5 +/- 0.2 mm Hg/ml/min, p < 0.05), but not in group AC. Between these ages, minimal coronary vascular resistance per unit heart weight increased to a greater extent in group AC (from 2.9 +/- 0.2 to 7.0 +/- 0.3 mm Hg/ml/min/gm, p < 0.05) than in group C (from 1.8 +/- 0.0 to 2.7 +/- 0.1 mm Hg/ml/min/gm, p < 0.05). In the second study, hearts were arrested with St. Thomas' Hospital cardioplegic solution (15 degrees C) and subjected to hypothermic (15 degrees C) global ischemia (210 minutes at 3 weeks and 180 minutes at 6 weeks) followed by 65 minutes of reperfusion. At 3 weeks of age, coronary resistance profiles, changes in left ventricular end diastolic pressure, and recoveries of left ventricular developed pressure during reperfusion were similar in groups C and AC. At 6 weeks of age, however, group AC had greater coronary resistance during reperfusion, a greater increase in left ventricular end-diastolic pressure (28 +/- 5 versus 1 +/- 1 mm Hg, p < 0.05), and consequently a reduced recovery of left ventricular developed pressure (50.7% +/- 3.6% versus 75.9% +/- 4.0%, p < 0.05) relative to group C. Thus 2 weeks of left ventricular pressure overload during neonatal development in the rat has little effect on coronary reserve and ischemic tolerance. Prolonged (5 weeks) pressure overload, however, results in postischemic diastolic dysfunction, which may be due to perfusion abnormalities during reperfusion. PMID- 8419693 TI - Mechanical durability of pulmonary allograft conduits at systemic pressure. Angiographic and histologic study in lambs. AB - We examined the mechanical durability of cryopreserved pulmonary allograft conduits at systemic pressure in lambs. Composite valveless tube grafts made of cryopreserved pulmonary allograft and a length of Dacron tube were implanted in the thoracic aortic position in 10 lambs (aged 1 month, mean weight 11.4 kg). The pulmonary allografts were harvested from similar-sized lambs (aged 1 month, mean weight 10.5 kg) at 0 to 48 hours postmortem and were cryopreserved. Recipient sheep were catheterized at 1 week (baseline) and at 1, 4, 6, 9, and 12 months postoperatively to allow changes in the diameters of the grafts to be followed up by angiography. Samples of the grafts were examined histologically at the time of harvest, at implantation after cryopreservation and thawing, and at the end of the study. At the postoperative baseline study, the pulmonary allografts had almost doubled (mean 21.1 mm) their implantation diameters (unpressurized mean 12.6 mm). During the 12-month study period, the allografts further increased their angiographic diameter by 36.2% compared with baseline (p < 0.01). One animal died at 3 1/2 months postoperatively and was found to have an aneurysm of the allograft. Five of the remaining eight lambs showed aneurysmal dilatation (more than 40% increase in diameter relative to pressurized baseline) at 12 months. Postmortem study revealed disruption at the suture line between allograft and Dacron in these five recipients. Histologic study showed much less calcification than was seen in our previous study of aortic allografts. There was loss of most of the smooth muscle, intimal and adventitial fibrous proliferation, and occasional dense lymphocytic infiltrates associated with foci of persistent smooth muscle cells. Time of harvest after donor death did not affect the histologic appearance after 12 months' implantation. Pulmonary allografts dilate substantially at systemic pressure. This characteristic has the benefit of allowing progressive enlargement with time as a recipient grows but also carries the risk of aneurysm formation, particularly pseudoaneurysm at the suture line when anastomosed with a noncompliant Dacron prosthesis. PMID- 8419694 TI - Is there an anatomic basis for subvalvular right ventricular outflow tract obstruction after an arterial switch repair for complete transposition? A morphometric study and review. AB - The study was initiated by reports on right ventricular outflow tract obstruction in complete transposition of the great arteries after an arterial switch repair. We investigated 39 heart specimens with native, unoperated transposition of the great arteries. Of these, 14 hearts had a ventricular septal defect; 25 had an intact ventricular septum. In each heart specimen the narrowest site of the subaortic outflow tract was measured and compared with the circumference of the aortic orifice. Obstruction was considered to be present if the outflow tract circumference was less than that of the aortic orifice. In addition, the diameter of the ascending aorta immediately above the level of the valve orifices was measured and compared with that of the pulmonary trunk. An obstruction was present in the subaortic right ventricular outflow tract of two hearts (5.1%): one of the obstructions, in a neonatal heart with intact ventricular septum, was caused by a prominent supraventricular crest and anterior trabeculations; the other obstruction was an additional extensive muscular hypertrophy, in the heart of a 13-year-old patient with a similar anatomy, and a septal defect. A mismatch between the diameters of the ascending aorta and the pulmonary trunk was present in 15 of 32 hearts measured. Our observations and a review of the literature confirm that subvalvular right ventricular outflow tract obstruction in hearts with native transposition of the great arteries is infrequent. Nevertheless, the anatomic characteristics of the right ventricular outflow tract are such that the tract is intrinsically narrow and muscular hypertrophy may easily lead to obstruction. After an arterial switch operation, subvalvular obstruction could be caused by dynamic processes analogous to those observed after relief of isolated pulmonary valve stenosis. Anatomic subvalvular obstruction could be due to either an obstruction that was not identified before operation or (a purely speculative hypothesis) subtle degrees of mismatch in size between the proximal aorta and the pulmonary trunk, which may be considered irrelevant at time of operation but may also set into pace a process of ongoing adaptive infundibular hypertrophy. PMID- 8419695 TI - Successful restoration of cell-mediated immune response after cardiopulmonary bypass by immunomodulation. AB - The objectives of this prospective randomized trial were to quantify immunosuppressive effects of cardiopulmonary bypass, to identify mechanisms responsible for postoperative immunosuppression, and to investigate the effects of immunomodulatory intervention on these mechanisms. Sixty patients were studied after cardiopulmonary bypass. Immunomodulatory therapy consisted of the cyclooxygenase inhibitor indomethacin, which blocks the downregulating agent prostaglandin E2, and thymopentin, which enhances T-lymphocytic activity. Twenty patients each received indomethacin either alone or combined with thymopentin. Twenty patients served as the control population. Our in vitro studies showed a decrease of CD4+ helper/inducer T cells and interleukin-2 receptor expression on T lymphocytes, while CD8+ suppressor/cytotoxic T cells and monocytes increased. Additionally, a depression of interleukin-1 and interleukin-2 synthesis as well as concurrent low gamma-interferon serum concentrations could be documented. These results indicate a downregulation of cell-mediated immune response. As an in vivo correlate of the immunomechanistic alterations, patients demonstrated an impaired delayed-type hypersensitivity response to an antigen skin test battery. These changes in immunoreactivity could be successfully counteracted by the combined immunomodulatory regimen, whereas sole indomethacin treatment could only partially restore depressed host defense parameters. With this study we could demonstrate for the first time that human lymphocytic interleukin-2 synthesis, which represents the key event among forward regulatory immune mechanisms, can be protected via in vivo immunoaugmentatory therapy and that this therapy can successfully counteract immunosuppressive effects of cardiopulmonary bypass. PMID- 8419696 TI - Effects of fixation back pressure and antimineralization treatment on the morphology of porcine aortic bioprosthetic valves. AB - Calcification frequently causes failure of porcine aortic valve bioprostheses; changes in collagen configuration induced by high-back-pressure fixation have deleterious effects on porcine aortic valve mechanics. Although modified porcine aortic valve bioprostheses that include the use of lower-pressure fixation and antimineralization treatments are used clinically, the morphologic characteristics of these valves are not known. We evaluated, by light and scanning and transmission electron microscopy, the comparative structural details of clinically processed Hancock Standard (no antimineralization treatment, 80 mm Hg fixation), Hancock II (T-6 antimineralization treatment, fixed initially at 1.5 mm Hg, then 80 mm Hg) and Intact (toluidine blue antimineralization treatment, zero-pressure-fixed [0 mm Hg]) porcine aortic valve bioprostheses as well as true low-pressure (1.5 mm Hg) fixed valves, zero-pressure-fixed porcine aortic valves (with no further treatment), and freshly fixed porcine aortic valve cusps as controls. Commercially processed valves had near-complete loss of endothelium and amorphous extracellular matrix and autolytic changes in the cuspal connective tissue cells. Both 80 and 1.5/80 mm Hg fixed valves, but not zero-pressure-fixed cusps (Intact valves, zero-pressure-fixed porcine aortic valves or immediately fixed porcine aortic valve cusps), had overall flattening and compression with near-complete loss of transverse cuspal ridges and collagen crimp; valves fixed at 1.5 mm Hg had intermediate features. T-6 and toluidine blue treatments induced no definite incremental microscopic changes attributable to the antimineralization treatment. No degenerative changes in collagen were noted in any valves that underwent antimineralization treatment. These studies indicate that valves fixed at zero but not at 80 or 1.5/80 mm Hg pressure retain collagen architecture virtually identical to that of relaxed native porcine aortic valve cusps and that the antimineralization treatments studied do not adversely affect collagen morphology. PMID- 8419697 TI - Clinical significance of epicardial pacing wire cultures. AB - Routine cultures of epicardial pacing wires removed 5 to 10 days postoperatively were obtained in 205 adults who underwent cardiac operations through median sternotomy. The study was conducted in a double-blind prospective fashion in which clinicians were unaware of culture results. With the exception of 10 out-of town patients who were followed up only until the day of hospital discharge, the patients were followed for at least 6 weeks (195 patients) for evidence of poststernotomy wound infections. Deep wound infection rate was slightly less than 1% in this patient population, with less than 0.5% having had superficial wound problems. Of the 205 patients, 27 had positive epicardial pacing wires cultures, with a total of 30 microbial isolates. Of 30 isolates, 26 were consistent with local skin flora (Staphylococcus, Streptococcus, Enterococcus, and diphtheroids). Wound infection developed in none of these patients. The remaining four cultures were of either Enterobacter or Serratia. In two of these four patients deep sternal infections developed. In the remaining 178 patients whose wire cultures were negative, no deep sternal infections developed. The fact that all clinically manifested deep sternal infections were associated with positive epicardial pacing wires cultures suggests that epicardial pacing wires cultures may be useful in the treatment of high-risk patients or of those in whom deep sternal infections are suspected. PMID- 8419698 TI - Clinical experience with the Omnicarbon prosthetic heart valve. AB - The Omnicarbon prosthetic heart valve (Medical Inc., Inver Grove Heights, Minn.) was implanted in 124 patients (mean age 53 +/- 11 years); 66 of them had aortic valve replacement, 40 had mitral valve replacement, and 18 had both aortic and mitral valve replacement. Preoperatively 76.6% were in New York Heart Association class 3 or 4, and 84.7% were in class 1 or 2 after the operation. There were six (4.8%) early deaths and seven late deaths. Survival was 85% +/- 6% at 6 years in the aortic valve replacement group, 94% +/- 4% at 3 years in the mitral valve replacement group, and 78% +/- 11% at 4 years in the double valve replacement group. Freedom from cardiac death was 89% +/- 4% at 6 years (2.0% per patient year) in the aortic valve replacement group, 94% +/- 4% at 3 years (1.8% per patient-year) in the mitral valve replacement group, and 78% +/- 11% at 4 years (5.7% per patient-year) in the double valve replacement group. There were six valve-related complications. Freedom from valve-related complications was 92% +/- 4% at 6 years (1.5% per patient-year) in the aortic valve replacement group, 97% +/- 3% at 3 years (1.8% per patient-year) in the mitral valve replacement group, and 83% +/- 11% at 4 years (5.7% per patient-year) in the double valve replacement group. Cerebral hemorrhage was seen in two patients in the aortic valve replacement group. Freedom from all events was 80% +/- 7% at 6 years in the aortic valve replacement group, 88% +/- 6% at 3 years in the mitral valve replacement group, and 78% +/- 11% at 4 years in the double valve replacement group. Elevation of the postoperative serum lactate dehydrogenase levels was minimal in all groups. The maximum opening angle at rest was 60.0 +/- 8.9 degrees in the aortic position and 54.1 +/- 6.6 degrees in the mitral position. In conclusion, the Omnicarbon prosthesis had excellent postoperative clinical status and negligible hemolysis. PMID- 8419699 TI - Invited letter concerning: The importance of pulsatile flow when systemic venous return is connected directly to the pulmonary arteries. PMID- 8419700 TI - Reply to invited letter concerning: Changes in plasma-free thyroid hormones during cardiopulmonary bypass (J Thorac Cardiovasc Surg 1992;104:526-7) PMID- 8419701 TI - Cytoimmunologic monitoring for rejection and infection after lung transplantation. PMID- 8419702 TI - Should adenosine continue to be ignored as a cardioprotective agent in cardiac operations? PMID- 8419703 TI - Invited letter concerning: Transplantation tolerance and transplantation. PMID- 8419704 TI - Tetralogy of Fallot with pulmonary atresia, coronary artery-pulmonary artery fistula, and origin of left pulmonary artery from descending aorta: total correction in infancy. PMID- 8419705 TI - Effect of cardiopulmonary bypass on systemic release of neutrophil elastase and tumor necrosis factor. AB - Leukocyte counts, plasma neutrophil elastase, tumor necrosis factor-alpha and C reactive protein were determined serially in 19 patients undergoing elective coronary artery surgery with cardiopulmonary bypass. Neutrophil counts (mean +/- standard deviation 3.85 +/- 1.20 x 10(9)/L preoperatively) peaked 4 hours postoperatively at 10.35 +/- 4.24 x 10(9)/L (p < 0.001) and remained significantly elevated 48 hours postoperatively at 7.80 +/- 2.70 x 10(9)/L, p < 0.05. Plasma neutrophil elastase level (187 +/- 74 ng/ml preoperatively) peaked at 698 +/- 323 ng/ml at the end of surgery (p < 0.001) and remained significantly elevated at 424 +/- 146 ng/ml 48 hours postoperatively (p < 0.01). Peak elastase levels correlated significantly with duration of bypass (r = 0.47, n = 19, p < 0.05). Monocyte counts (0.29 +/- 0.19 x 10(9)/L preoperatively) peaked 4 hours postoperatively (0.87 +/- 0.41 x 10(9)/L, p < 0.001) and fell to baseline levels by 48 hours postoperatively. Plasma tumor necrosis factor-alpha, detectable in 10 of the 19 patients preoperatively (median 0.39 U/ml, range up to 10.1 U/ml), did not change significantly during or after bypass. Plasma C-reactive protein level (median 1.67 [range 0.69 to 34.33] micrograms/ml preoperatively) rose significantly to 3.99 (range 1.95 to 12.55) micrograms/ml 4 hours postoperatively (p < 0.01) and rose 48 hours postoperatively at 303 (210 to 410) micrograms/ml, p < 0.001. Oxygenation, determined by the respiratory index, was impaired at the end of operation (2.07 +/- 0.82) and remained impaired 24 hours postoperatively (2.48 +/- 0.83). Impairment of oxygenation was temporally related to elevated elastase levels, but neither peak elastase levels nor the change in elastase levels with lung reperfusion correlated significantly with the area under the respiratory index curve up to 6 hours postoperatively. This study demonstrates neutrophil elastase release during cardiopulmonary bypass but fails to show a definite role for neutrophil activation or tumor necrosis factor-alpha in the etiology of pulmonary dysfunction after cardiopulmonary bypass. PMID- 8419706 TI - Changes in plasma fibronectin in children after elective repair of congenital heart defects. AB - Plasma fibronectin is an attachment protein important for maintaining capillary integrity and host defense mechanisms. Depletion of plasma fibronectin has been shown to occur in adults after septic shock, major trauma, and burns. Limited laboratory and clinical studies suggest a correlation between decreased plasma fibronectin levels and increased pulmonary capillary permeability and tissue perfusion. Mild and transient plasma fibronectin depletion has been observed in adults after cardiovascular operations. We measured plasma fibronectin by immunoturbidometric assay in 20 children (age 6 months to 12 years) undergoing repair of congenital heart defects. Plasma fibronectin levels immediately after operations and daily thereafter were compared with the preoperative values. Plasma fibronectin declined on postoperative days 1, 2, 3, 4, and 5 (p < 0.05). A nadir was reached on day 3 with a tendency toward recovery thereafter. Patients with a therapeutic intervention score of more than 35 had greater magnitude of plasma fibronectin decline than those with a score of less than 35 at 24 hours after the operation (p < 0.005). We conclude that (1) significant and prolonged plasma fibronectin depletion occurs after cardiovascular operations in children; and (2) postoperative plasma fibronectin depletion is associated with increasingly complex surgical intervention. Reduced plasma fibronectin synthesis and more extensive operations for congenital heart defects are likely reasons for children being more susceptible than adults to plasma fibronectin depletion after cardiovascular operations. PMID- 8419707 TI - Recovery of postischemic contractile function is depressed by antegrade warm continuous blood cardioplegia. AB - To assess the effectiveness of warm antegrade continuous blood cardioplegia in the setting of an acute coronary arterial occlusion, we instrumented 19 Yorkshire swine to quantitate left ventricular global, systolic, diastolic, and regional mechanics. Data were acquired before and after 10 minutes of mid-left anterior descending coronary artery occlusion followed by 60 minutes of aortic crossclamping. Cardiac arrest was induced by the antegrade infusion of 20 ml/kg of warm (37 degrees C) or cold (4 degrees C) oxygenated blood cardioplegic solution followed by either continuous warm (75 ml/min, n = 9) or intermittent cold (10 ml/kg every 20 minutes, n = 10) cardioplegic reinfusions. Left anterior descending coronary artery occlusion was released 20 minutes after aortic crossclamping and resulted in warm-arrested hearts developing a 139% increase in global oxygen consumption compared with values obtained with the left anterior descending coronary artery occluded (p < 0.02). Recovery of global left ventricle contractility, quantitated by the linear preload recruitable stroke-work relationship, was significantly worse after warm cardioplegia (52.4% +/- 5.1% versus 68.0% +/- 5.9%, warm versus cold, p < 0.05). Similarly, left anterior descending coronary artery regional ischemic zone contractility recovered 34.5% +/- 7.3% of control function with cold cardioplegia, whereas warm cardioplegia resulted in -11.36% +/- 7.46% functional recovery indicative of dyssynchronous contraction (p < 0.05). Diastolic compliance, calculated with an exponential end diastolic pressure-versus-volume relationship, was not changed postischemically in either group. These data suggest that warm antegrade blood cardioplegia may potentiate acute ischemic injury and provide inadequate myocardial protection. PMID- 8419708 TI - Warm versus cold blood cardioplegia--is there a difference? AB - This experimental study sought to compare the effectiveness of warm blood cardioplegia versus cold blood cardioplegia in protecting areas of ischemic myocardium during urgent coronary revascularization. In 40 adult pigs, the second and third diagonal vessels were occluded with snares for 90 minutes. All animals were then placed on cardiopulmonary bypass and underwent 45 minutes of cardioplegic arrest followed by 3 hours of reperfusion during which time the coronary snares were released. During the period of cardioplegic arrest, 10 pigs received antegrade continuous warm blood cardioplegic solution (37 degrees C) at 100 ml/min; 10 animals received retrograde warm blood cardioplegic solution at 100 ml/min; 10 received intermittent, antegrade cold blood cardioplegic solution (4 degrees C), and 10 animals received intermittent, antegrade/retrograde cold blood cardioplegic solution. Hearts protected with antegrade warm blood cardioplegic solution had the lowest pH values in the area at risk (6.59 +/- 0.10 antegrade warm blood cardioplegia versus 6.80 +/- 0.10 retrograde warm blood cardioplegia versus 6.72 +/- 0.18 antegrade cold blood cardioplegia versus 6.85 +/- 0.15 antegrade/retrograde cold blood cardioplegia and the highest area of necrosis (42% +/- 3% antegrade warm blood cardioplegia versus 26% +/- 2% [p < 0.05 from antegrade warm blood cardioplegia] retrograde warm blood cardioplegia versus 31% +/- 2% [p < 0.05 from antegrade warm blood cardioplegia] antegrade cold blood cardioplegia versus 21% +/- 2% [p < 0.05 from antegrade warm blood cardioplegia] antegrade/retrograde cold blood cardioplegia). We conclude that in the presence of an acute coronary occlusion with ischemic myocardium, warm blood cardioplegic solution should be given in a continuous retrograde fashion and does not result in myocardial protection superior to the protection that can be achieved with antegrade/retrograde cold blood cardioplegic solution. PMID- 8419709 TI - Impaired endothelium-dependent coronary microvascular relaxation after cold potassium cardioplegia and reperfusion. AB - Myocardial dysfunction after cardiac operations might be influenced by altered myocardial perfusion in the postoperative period. To investigate possible alterations in vascular reactivity, in vitro coronary microvascular responses were examined after ischemic cardioplegia with use of a porcine model of cardiopulmonary bypass. Since myocardial perfusion is primarily regulated by arteries less than 200 microns in diameter, these vascular segments were examined. After 1 hour of ischemic arrest with cold crystalloid cardioplegia and 1 hour of reperfusion, microvessels (100 to 190 microns in diameter) were pressurized in a no-flow state, preconstricted by 30% to 60% of the baseline diameter with acetylcholine, and examined with video microscopic imaging and electronic dimension analysis. Endothelium-dependent relaxations to bradykinin (55% +/- 13% versus 99% +/- 1% = maximum relaxation of the preconstricted diameter in cardioplegia-reperfusion vessels versus control vessels, respectively; p < 0.05) and the calcium ionophore A 23187 (33% +/- 6% versus 90% +/- 4%; p < 0.05) were markedly impaired while endothelium-independent relaxation to sodium nitroprusside was similar to control value. After 1 hour of ischemic cardioplegia without reperfusion, endothelium-dependent relaxation was only slightly affected. Transmission electron microscopy showed minimal endothelial damage after ischemic cardioplegia and reperfusion. These findings have important implications regarding coronary spasm and cardiac dysfunction after cardiac operations. PMID- 8419710 TI - Effects of high plasma epinephrine and Ca2+ concentrations on neonatal myocardial function after ischemia. AB - Administration of catecholamines to newborn infants may potentiate reperfusion injury because of increased transsarcolemmal Ca2+ influx and the presence of less developed sarcoplasmic reticulum in the immature hearts. We investigated the effect of administration of epinephrine (1.5 micrograms/kg per minute for 120 minutes) before ischemia and modified serum ionized Ca2+ concentrations in the cardioplegic solution and perfusate on postischemic left ventricular systolic and diastolic function in 25 piglets (5 to 7 days old) undergoing 90 minutes of cold blood cardioplegic arrest. The piglets were divided into four groups; Ca2+ 1.2 mmol/L, group A (n = 6), Ca2+ 0.25 mmol/L, group B (n = 6), Ca2+ 1.2 mmol/L and epinephrine, group C (n = 6), Ca2+ 0.25 mmol/L and epinephrine, group D (n = 7). Left ventricular function was assessed by a conductance catheter in the left ventricle measuring end-systolic and end-diastolic pressure-volume relationships during transient vena caval occlusion. By analysis of covariance, only Ca2+ concentration was important in predicting ventricular function recovery after ischemia (p < 0.01). End-systolic elastance decreased in all groups after ischemia; the magnitude was significantly greater in the normal groups (51% versus 35%, p < 0.01). There was a significant increase in the chamber stiffness index after administration of epinephrine before ischemia (p < 0.05). Groups with low Ca2+ perfusate (B and D) had no change in chamber stiffness index after ischemia. In contrast, there was a significant increase in chamber stiffness in the normal Ca2+ groups with (C) or without (A) epinephrine after ischemia (p < 0.05). Adenosine triphosphate stores declined significantly in the normal Ca2+ groups--48% versus 18% in the low Ca2+ groups (p < 0.01). We conclude that low Ca2+ concentrations in the perfusate and cardioplegic solutions better preserve left ventricular function in the normal and in epinephrine-stressed neonatal heart after ischemia. PMID- 8419711 TI - Skeletal muscle-powered ventricle. Effects of size and configuration on ventricular function. AB - The optimal size and configuration of skeletal muscle-powered ventricles are still undetermined. This study was aimed at comparing three types of skeletal muscle-powered ventricle: (A) a small size (15 ml capacity) double-layered pump, (B) a small size (15 ml capacity) single-layered pump, and (C) a large size (40 to 60 ml capacity) single-layered pump constructed sequentially with the same untrained latissimus dorsi muscle of 12 mongrel dogs. The skeletal muscle-powered ventricle was connected to a mock circulation system, the stroke volumes against 40 to 160 mm Hg of afterload at 5 to 60 mm Hg of preload were measured, and the stroke work was computer analyzed on line. Raising the preload from 5 to 60 mm Hg increased the peak isovolumic developed pressure (A) from 91.3 +/- 11.0 to 215.6 +/- 26.1 mm Hg, (B) from 92.8 +/- 12.0 to 166.3 +/- 19.0 mm Hg, and (C) from 32.3 +/- 5.2 to 121.4 +/- 15.5 mm Hg (p < 0.05, C versus A and B). Similarly, the stroke volume (stroke work) against an afterload of 120 mm Hg increased (A) from 3.8 +/- 0.5 ml (0.22 +/- 0.04 x 10(6) ergs) to 14.5 +/- 1.1 ml (1.05 +/- 0.11 x 10(6) ergs), (B) from 4.5 +/- 0.7 ml (0.30 +/- 0.08 x 10(6) ergs) to 10.7 +/- 0.9 ml (0.63 +/- 0.08 x 10(6) ergs), and (C) from 1.8 +/- 0.5 ml (0.09 +/- 0.04 x 10(6) ergs) to 24.0 +/- 3.6 ml (1.94 +/- 0.41 x 10(6) ergs) (p < 0.05, C versus B at 5 mm Hg of preload; p < 0.05, C versus A and B at preloads > or = 30 mm Hg). At low preloads (5 to 15 mm Hg) both small pumps generated a significantly larger stroke volume (stroke work) than the large pump, whereas at high preloads (> or = 30 mm Hg) the large pump generated a significantly larger stroke volume (stroke work) than the small pumps. It is concluded that under physiologic preload, B (small single-layered pump) performs better than or at least as well as A (small double-layered pump) and C (large single-layered pump), despite being constructed with only one half of the muscle mass used for either A or C. PMID- 8419712 TI - Tranexamic acid (Cyklokapron) is not necessary to reduce blood loss after coronary artery bypass operations. AB - The contribution of fibrinolysis to postoperative bleeding after cardiopulmonary bypass led to routine use of tranexamic acid, a potent antifibrinolytic drug, for a period of time. Two hundred patients undergoing elective coronary artery bypass operations were studied, one group of 100 patients given tranexamic acid (40 mg/kg) (group I) after bypass and one subsequent group of 100 patients (group II) serving as a control group. All patients were treated by the same team, and the groups were comparable in all major clinical parameters. The mean mediastinal drainage in group I was 565 +/- 239 ml versus 656 +/- 257 ml in group II. Univariate and multivariate analysis revealed statistical significance (p = 0.02) when corrected for body surface area. However, applying a consistent blood conservation protocol, including removal of autologous blood before bypass for retransfusion after bypass, returning of all oxygenator and tubing contents to the patients, and autotransfusion of the mediastinal shed blood up to 18 hours postoperatively, resulted in nearly identical hemoglobin concentration at discharge (119 +/- 14 gm/L in group I and 121 +/- 14 gm/L in group II). The prevalence of postoperative myocardial infarction included five patients in group I compared with one patient in group II. Although not statistically significant (p = 0.2), the difference is of concern. Tranexamic acid has a beneficial effect on reducing postoperative bleeding after coronary artery bypass operations. The routine use of the drug is not recommended, however, because its effect is a weak one, and it may be of potential hazard by precipitating thrombosis and eventual myocardial infarction. PMID- 8419713 TI - Thoracoscopic pleurectomy for treatment of complicated spontaneous pneumothorax. AB - This report describes a thoracoscopic approach for performing parietal pleurectomy. We have developed and used this technique successfully in 12 patients for treatment of recurrent spontaneous pneumothorax with extended bullous lung alterations (stage 4 according to the classification of Vanderschueren). For this purpose we need videoendoscopy and specially designed equipment, including pliable silicone trocars and angled instruments. The mean age of the patients was 38 years; no deaths and no complications occurred. The average period of postoperative hospitalization was 3.3 days. During the follow up period ranging between 5 and 10 months (mean 7.5), no relapsing pneumothorax was observed. PMID- 8419714 TI - Medical tumors of the chest wall. Solitary plasmacytoma and Ewing's sarcoma. AB - Primary solitary plasmacytoma and Ewing's sarcoma of the chest wall are relatively uncommon tumors, and data concerning treatment and results are sparse. To assess the results of therapy we reviewed our 40-year experience. METHODS: Records of 24 patients with solitary plasmacytoma and 62 with Ewing's sarcoma arising in the chest wall who were admitted to our institution from 1949 to 1989 were reviewed. RESULTS: In the group with plasmacytoma (n = 24), ages ranged from 35 to 75 years (median 59 years); male/female ratio was 2.4:1. The presenting complaint was pain or mass or both in 92% (22/24). Primary therapy was local only in 5 (resection in 3, radiotherapy in 2), chemotherapy in 16 (resection in 5, radiotherapy in 10, and chemotherapy alone in 1); 3 patients did not receive therapy. Multiple myeloma developed subsequently in 75% (18/24). Overall 5-year survival was 38% (median 56 months). Age, sex, site of primary tumor, and local therapy did not significantly impact on survival. Ages in the patients who had Ewing's sarcoma (n = 62) ranged from 2 to 39 years (median 16 years); male/female ratio was 1.6:1. Presenting complaint was pain or mass or both in 98% (61/62). Primary therapy was local in 17 (resection in 7, radiotherapy in 7, resection plus radiotherapy in 3) and chemotherapy in 45 (plus resection in 29, resection and radiotherapy in 10, and radiation therapy alone in 3). Overall 5-year survival was 48% (median 57 months). Age, sex, and site of primary tumor did not significantly impact on survival. Patients in whom distant metastases developed (n = 48) had a significantly decreased survival (5 year, 28%) compared with those who did not have metastases (n = 14; 5 year, 100%). CONCLUSION: Plasmacytoma of the chest wall, even if solitary at presentation, should be considered a systemic disease, and therapy should be directed as such. For Ewing's sarcoma, although resection or radiotherapy may offer local control, because of the prevalence of distant metastases (77%), systemic therapy should be considered an integral part of treatment. PMID- 8419715 TI - Lung size matching for double lung transplantation based on the submammary thoracic perimeter. Accuracy and functional results. The Joint Marseille-Montreal Lung Transplant Program. AB - The present study evaluates the accuracy of submammary thoracic perimeter for lung size matching between donor and recipient and analyzes the influence of donor lung size discrepancies on functional outcome after double lung transplantation. The population is composed of 18 double lung graft recipients, 16 of whom had cystic fibrosis. The lung size match was assessed by comparison of predicted total lung capacity of donor and recipient: five patients were matched in a 10% confidence interval; four received smaller lungs, and nine received larger ones. The functional outcome was assessed with the spirometric values measured at 3 and 6 months after transplantation. The final functional result was not influenced by the lung size (r = 0.142 for total lung capacity; r = 0.372 for vital capacity; r = 0.378 for forced expiratory volume in 1 second). For larger lungs the final result tended to the recipient's predicted, whereas for smaller lungs, spirometry tended to the donor's predicted (r = 0.906 for total lung capacity; r = 0.875 for vital capacity; r = 0.874 for forced expiratory volume in 1 second). The thoracotomy effect, that is, restrictive syndrome at 3 months that resolves at 6 months, was not correlated with the lung size (r = 0.07 for total lung capacity; r = 0.436 for vital capacity). It is concluded that respiratory functional result is not affected by larger lungs; despite the wide range of error, the submammary thoracic perimeter appeared to be a satisfactory selection parameter in this group of patients. PMID- 8419716 TI - Neuropathies associated with diabetes. AB - Diabetic neuropathy is the most frequent complication of diabetes and the leading cause of polyneuropathy in the Western world. A distal symmetric predominantly sensory polyneuropathy is the most common of the diverse neuropathies that occur secondary to diabetes. Pain is often the most bothersome and difficult to treat symptom of diabetic neuropathy. Autonomic neuropathy is a frequent feature of diabetic neuropathy and the source of many significant problems including postural hypotension, gastroparesis, diarrhea, constipation, neurogenic bladder, and male impotence. Physicians need to be familiar with the multiple, less common forms of diabetic neuropathy, as these often mimic other medical or neurologic conditions. The cause of diabetic neuropathy is not determined, but abundant evidence suggests that both metabolic and ischemic nerve injury are likely factors. These should not be considered mutually exclusive causes of diabetic neuropathy as both factors likely operate to different degrees to produce the clinical spectrum of neuropathies that are seen in diabetes. Although no effective treatment exists to cure diabetic neuropathy, improvement is possible with glycemic control and symptomatic therapy. PMID- 8419717 TI - Headache to worry about. AB - The headache to worry about is one that is unique or different from headaches which the patient has suffered in the past. The association with the headache of meningismus or of focal neurologic symptoms of oculoparesis, other cranial nerve palsies, hemiparesis, or loss of consciousness are particularly worrisome, especially if onset is recent and acute. Headaches related to arteritis or vasculitis usually have a slower subacute course but may also produce focal neurologic deficits. For subjects over 50 years old, temporal arteritis is always a consideration and any new type of headache requires testing of the sedimentation rate to rule out this treatable but potentially devastating problem. PMID- 8419718 TI - Update on epilepsy. AB - Increasing knowledge of prognostic factors and of treatment outcomes has led to major changes in the treatment of epileptic seizures. The management of first seizures can now be individualized, based on the probability of seizure recurrence and its expected repercussions. When a decision is made to treat initial or recurrent seizures, monotherapy with nonsedating AEDs is recommended. Therapy should also be optimized to achieve seizure control as early as possible. During AED therapy, clinical monitoring of drug reaction symptoms is preferred over routine laboratory testing. Laboratory monitoring is not cost-effective, and there is no proof that it can forecast drug reactions. The risk of congenital malformations in newborns prenatally exposed to AEDs is around 5%, which is 2 to 2.5 times that of the general population. Valproate and carbamazepine have recently been associated with a 1% risk of spina bifida. There are measures that can be used to reduce the risk of teratogenicity, the most important of which is using the least amount of AEDs necessary to control seizures during pregnancy. Although AEDs to reduce the efficacy of oral contraceptives, oral contraceptives are still reliable with certain precautions. For patients with intractable epilepsy, surgery should be considered to improve seizure control. Proper selection of candidates increases the probability of an excellent outcome in these patients, who otherwise have no hope of leading a normal life. Postsurgical outcome largely depends on the location, the extent, and the pathology of the epileptogenic tissues in the brain. PMID- 8419719 TI - Reversible dementias. AB - With so many conditions that can manifest in dementia, the question arises as to how extensive an evaluation need be done on the individual patient presenting with dementia. Thorough physical, neurologic, and psychiatric examinations are the cornerstones of the work-up, with special attention paid to the history, use of medications, and mental status of the patient. Laboratory tests recommended by the National Institutes of Health Consensus Conference on Differential Diagnosis of Dementing Diseases include a complete blood count, electrolyte and metabolic screen, thyroid panel, vitamin B12 and folate levels, syphilis serology, urinalysis, chest radiograph and electrocardiogram, and head CT scan. These evaluations are sufficient to diagnose the majority of treatable dementias. Other evaluations including magnetic resonance imaging, electroencephalography, cerebrospinal fluid examination, cerebral blood flow and metabolism measures (rate of cerebral blood flow, single photon emission computed tomography, and positron-emission tomography), and brain biopsy all can be of additional assistance in diagnosing the cause of the dementia when justified by the clinical setting. When the appropriate diagnosis is made, therapy is directed at the primary disorder. Successful treatment of the primary condition may result in stabilization or partial or complete reversal of the cognitive disturbance. In some instances, judicious pharmacologic management of the accompanying behavioral disturbance may be required. PMID- 8419720 TI - Viral encephalitis. AB - Viral encephalitis represents an important source of morbidity and mortality worldwide. Numerous viruses possess neurovirulence, producing encephalitic disorders that usually consist of fever, headache, vomiting, altered consciousness, focal or generalized seizures, and motor dysfunction. Contemporary virologic methods frequently allow rapid and specific identification of viral pathogens, but the etiologic agent remains uncertain in 25% or more of encephalitis patients. Although acyclovir substantially reduces mortality and improves outcome for patients with herpes simplex virus encephalitis, supportive care remains the only therapy available for most patients with virus encephalitis. PMID- 8419721 TI - Neurologic complications of thyroid and parathyroid disease. AB - Common thyroid and parathyroid disorders present with reversible neurologic signs and symptoms affecting the central and peripheral nervous system, musculature, and mental function. Patients with thyrotoxicosis may have myopathy, spasticity, seizures, and multiple psychiatric symptoms. A deficiency of thyroid hormone also causes muscle weakness and may be accompanied by reversible muscle hypertrophy or movement disorders. The chronic hypercalcemia that develops secondary to hyperparathyroidism produces many psychiatric and cognitive symptoms, as well as a reversible myopathy. Calcium deficiency leads to neuromuscular irritability, paresthesias, and tetany. Psychiatric disorders are also common in this disorder. PMID- 8419722 TI - Neurologic complications of systemic cancer. AB - Neurologic complications of systemic cancer are common. Metastatic complications include intracranial metastasis, spinal metastasis, leptomeningeal metastasis, and metastasis to peripheral nerves. Treatment is often effective in preventing further neurologic disability and pain. Early diagnosis is important. Nonmetastatic complications of systemic cancer include a wide variety of neurologic illnesses that can also occur in the noncancer population. The most common is toxic metabolic encephalopathy. Other nonmetastatic complications include cerebrovascular complications, complications of treatment, immunocompromised infections, and paraneoplastic syndromes. PMID- 8419723 TI - Medical complications of head injury. AB - There are many common and significant medical complications of head injury. These include (1) cardiovascular problems such as hyperdynamic state, myocardial injury, and dysrhythmias; (2) respiratory changes such as neurogenic pulmonary edema, hypoxia, abnormal ventilatory patterns, pulmonary infections, and pulmonary emboli secondary to deep vein thrombosis; (3) consumption coagulopathy; (4) water and electrolyte derangements--hypo- and hypernatremia; (5) hypothalamic/pituitary dysfunction--syndrome of inappropriate secretion of antidiuretic hormone and diabetes insipidus; (6) increased general metabolism with loss of immunocompetence, respiratory compromise, and complications of decreased activity; (7) gastrointestinal difficulties, particularly stress gastritis; and (8) infectious problems including those related to contamination from open wounds and foreign bodies such as monitors. PMID- 8419724 TI - Management of intracranial hypertension. AB - Intracranial hypertension is the final common denominator of morbidity and mortality for diverse neurologic problems, and its proper treatment requires the heuristic application of the available therapeutic alternatives when the clinical situation and patient's prognosis warrants treatment. The initial therapeutic focus for ICP reduction should be control of factors that may aggravate intracranial hypertension such as inappropriate head and body position, elevated body temperature, pain, noxious stimuli, elevated airway pressure, elevated blood pressure, seizures, and hypotonic intravenous fluids. The appropriate conventional therapies (e.g., hyperventilation, osmotic agents, sedatives, barbiturates, and cerebrospinal fluid removal) should be selected based on the details of each individual case. Surgical removal of intracranial mass lesions may be indicated in some circumstances, particularly for intractable intracranial hypertension and progressive, severe brain tissue shifts. PMID- 8419725 TI - Abnormalities of hemostasis in ischemic stroke. AB - Despite important new diagnostic laboratory and imaging technologies, the cause of brain infarction remains unexplained in 20% to 40% of subjects. Most stroke patients do not require extensive evaluations of coagulation, but hypercoagulability may account for a significant proportion of unexplained strokes. Hemostatic abnormalities associated with stroke may be broadly classified as familial or acquired. Principal among the familial thrombotic coagulopathies are deficiencies in concentration or function in protein-C, protein-S, and antithrombin III, but other hereditary abnormalities include sickle cell disease, homocystinuria, and dysfibrinogenemia. The acquired disorders of hemostasis associated with stroke probably constitute a larger proportion of the important stroke-related coagulopathies. In particular, the aPL antibody syndrome is now strongly associated with thrombotic events including stroke, although neither the mechanism of thrombosis nor effective therapies for this syndrome have been clearly elucidated. Many of the acquired hemostatic abnormalities exist within a special clinical setting such as with malignancy or with myeloproliferative diseases, nephrotic syndrome, and liver disease. Presumably many of these share common pathways of coagulation activation or dysfunction with the inherited disorders. Most of the hemostatic disorders in stroke are associated with dysfunction of vascular endothelium and abnormalities of or interference with the natural anticoagulant proteins: protein-C, protein-S, and antithrombin III. Improved understanding of these relationships should lead to better diagnosis and treatment for people at risk of stroke. PMID- 8419726 TI - Diabetes and stroke. AB - Diabetes is a major risk factor for development of ischemic cerebrovascular disease. Patients with diabetes are at least two times more likely to have a stroke than nondiabetics. In addition, they are more likely to suffer increased morbidity and mortality after stroke. The mechanism of production of stroke secondary to diabetes may be due to cerebrovascular atherosclerosis, cardiac embolism, or rheologic abnormalities. The evaluation of cerebrovascular disease in diabetic patients is similar to the nondiabetic patient, with particular attention paid to adequate hydration prior to the administration of contrast agents. Treatment options for stroke in diabetics requires individualization but should include risk factor modification, and may include platelet antiaggregants, anticoagulation, or, in a well-defined subgroup, carotid endarterectomy. PMID- 8419727 TI - Abnormalities of the neonatal ear: otoscopic observations, histologic observations, and a model for contamination of the middle ear by cellular contents of amniotic fluid. AB - It is unknown whether childhood ear disease could be present long before symptoms provoke an initial otoscopic examination. A newborn middle ear might or might not start in a pristine, privileged state. The clinician evaluating later infant and childhood ear disease is often unaware of the status of a patient's ear from the neonatal period, the earliest time at which the tympanic membrane can be evaluated. Adding to the physician's handicap, normative otoscopic and histologic data on the neonatal ear are incomplete. In order to test the hypothesis that disease in the neonatal middle ear may be more common than is generally appreciated, the population of critically ill neonates was selected for study since this group can provide both clinical as well as histologic data. This manuscript is divided into three parts. Clinically, otoscopic observations were analyzed on infants in an intensive care unit. Histologically, neonatal temporal bones were studied for normal anatomy and pathology of the middle ear and antrum. Experimentally, an animal study was performed to evaluate the potential effect of amniotic fluid cellular contents aspirated into the middle ear. I. Clinical Otoscopic Observations. Daily otoscopic examination was conducted on 44 neonates in an intensive care unit. Specific parameters of the otoscopic examination were evaluated to compare with the normal, translucent tympanic membrane of the older child. The otoscopic appearance was found to be abnormal in 97.7% of neonatal ears. Of the otoscopic parameters evaluated, right ears averaged 2.6 abnormalities and left ears averaged 2.5 otoscopic abnormalities. The otoscopic appearance of the neonate in the neonatal intensive care unit is nearly universally abnormal. II. Temporal Bone Histologic Observations. One hundred eleven temporal bones from 56 neonates were collected for histologic study by light microscopy. Mesenchyme filling more than 60% of the middle ear space was found in 13 bones. Amniotic fluid cellular content was detected in 90 bones. Purulent otitis media was detected in 24 bones. Varying amounts of blood were found in the middle ear space of 34 bones. Only 7 of the bones had no significant middle ear abnormality. It is concluded that in the critically ill neonate, the middle ear and antrum usually contain cellular or fluid material, often in significant volume, that would not be considered normal in the older patient. III. An Animal Model Simulating Contamination of the Middle Ear by Cellular Contents of Amniotic Fluid.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419728 TI - Comparison of pig cornea preservation solutions using quantitative 31P NMR spectroscopy. AB - We compared the abilities of three solutions (bicarbonate-free glucose-phosphate Ringer's solution (EP II), lactated Ringer's solution and physiologic saline solution) to preserve pig cornea by measuring phosphate metabolites with 31P NMR spectroscopy. EP II preserves corneal viability better than others, and preserving extracted cornea also is better than preserving the whole eye. PMID- 8419729 TI - An assessment of spin-echo rotating-frame imaging for spatially localized determination of short T2 relaxation times in vivo. AB - The rotating frame of localized spectroscopy can be augmented by the inclusion of a refocusing pulse to enable the measurement of T2 relaxation times. This technique is particularly appropriate for determining short relaxation parameters due to the absence of time consuming switched B(omicron) field gradients. We have evaluated the accuracy of this protocol by measuring localized T2s in the range of 1 to 20 ms. Preliminary data obtained from muscle and liver of normal and iron overloaded human subjects are also presented. PMID- 8419730 TI - A 1H-NMR method for determining temperature in cell culture perfusion systems. AB - This report describes a noninvasive 1H-NMR method for measuring absolute temperatures (+/- 0.2 degrees C) in biological samples and, in particular, in cell culture perfusion systems, utilizing the linear temperature dependence of the water chemical shift relative to the temperature-independent shift of one of the components of the biological medium, e.g., pyruvate, acetate or lactate. The effects of flow on temperature can be monitored and appropriate adjustment of the temperature controller can be made. PMID- 8419731 TI - A simple method of measuring gradient induced eddy currents to set compensation networks. AB - We describe a technique for measuring the time dependence and field distortions of magnetic fields due to eddy currents (EC) produced by time-dependent magnetic field gradients. The EC measuring technique uses a sample with short T1, T2 and many rf excitation pulses and free induction decays (FIDs) to measure the out-of phase component of the FIDs, which are proportional to gamma delta B, the amount by which the signal is off resonance. The measuring technique is sensitive, easy to implement and interpret, and useful for setting preemphasis compensation networks. PMID- 8419732 TI - 31P localized spectroscopy of fetal brain in utero. AB - 31-Phosphorus (31P) nuclear magnetic resonance (NMR) spectroscopy of fetal brain in utero was obtained entirely noninvasively in rat utilizing image selected in vivo spectroscopy (ISIS). The results were in excellent agreement with the data obtained using the surface coil method in the late stage fetus, namely, high phosphomonoester (PME), low phosphocreatine (PCr), and high intracellular pH. Localized spectroscopy provides unprecedented opportunities for the investigation of fetal brain metabolism in utero. PMID- 8419733 TI - Molecular self-diffusion of intracellular metabolites in rat brain in vivo investigated by localized proton NMR diffusion spectroscopy. AB - Molecular self-diffusion coefficients of water (0.75 +/- 0.05), N-acetylaspartate (0.27 +/- 0.04), creatines (0.27 +/- 0.04), and cholines (0.28 +/- 0.08) x 10(-5) cm2 s-1 were obtained from localized proton NMR spectra of rat brain in vivo using diffusion-weighted stimulated-echo (STEAM) sequences with a diffusion time of (delta--delta/3) = 17 ms. PMID- 8419734 TI - On the extracellular contribution to multiple quantum filtered 23Na NMR of perfused rat heart. AB - We investigated the contribution of extracellular Na+ to the multiple-quantum filtered 23Na NMR signal of perfused rat hearts to determine if the presence of shift reagent Dy(PPPi)2 and inorganic phosphate were somehow responsible for the generation of extracellular multiple quantum coherence. Neither phosphate nor shift reagent caused an increase in the total multiple-quantum filtered signal intensity or in the percent contribution from extracellular ions. On the contrary, addition of Dy(PPPi)2 actually decreased the total signal intensity from intra- and extracellular ions. Further addition of 1.5 mM Gd(PPPi)2 eliminated the extracellular contribution. These data indicate that the previously reported extracellular contribution in perfused hearts is a true contribution of extracellular ions, and not an artifact originating from their interaction with the shift reagent. PMID- 8419735 TI - Phase velocity mapping with a real time line scan technique. AB - A real-time, 20-Hz, one-dimensional MR velocity imaging technique is described. A two-dimensional RF pulse excites a 3-cm diameter column. Velocity maps are formed from the phase difference between successive flow encoded and compensated acquisitions. A three-point subtraction variation provides reduced sensitivity to static spins. PMID- 8419736 TI - Functional MRI of human brain activation at high spatial resolution. AB - Functional activation maps of the human visual cortex were obtained at a spatial resolution almost two orders of magnitude better than achievable by positron emission tomography and within measuring times of a few seconds. Transient alterations in the concentration of paramagnetic deoxyhemoglobin were conveniently detected at 2.0-T with use of RF-spoiled FLASH MRI sequences employing gradient echo times of 6 to 60 ms and voxel sizes of 2.5 to 39 microliters. PMID- 8419737 TI - The true T1 values of myocardial high-energy phosphates? PMID- 8419738 TI - The origin of biexponential T2 relaxation in muscle water. AB - Two theories have been proposed to explain the multiexponential transverse relaxation of muscle water protons: "anatomical" and "chemical" compartmentation. In an attempt to obtain evidence to support one or the other of these two theories, interstitial and intracellular macromolecular preparations were studied and compared with rat muscle tissue by proton NMR transverse relaxation (T2) measurements. All macromolecule preparations displayed monoexponential T2 decay. Membrane alteration with DMSO/glycerin did not eliminate the biexponential T2 decay of muscle tissue. Maceration converted biexponential T2 decay of muscle tissue to single exponential decay. It is concluded that the observed two component exponential T2 decay of muscle represents anatomical compartmentation of tissue water, probably intracellular versus extracellular. PMID- 8419739 TI - A three-dimensional spin-echo or inversion pulse. AB - In theory, multidimensional pulses can be designed to be selective in any number of dimensions. In practice, available gradient power has enforced a limit to two dimensions. We show here that three-dimensional pi pulses are feasible on commercial imaging machines provided that the range of off-resonance frequencies are limited. PMID- 8419740 TI - A high-field superferric NMR magnet. AB - Strong, extensive magnetic fringe fields are a significant problem with magnetic resonance imaging magnets. This is particularly acute with 4-T, whole-body research magnets. To date this problem has been addressed by restricting an extensive zone around the unshielded magnet or by placing external unsaturated iron shielding around the magnet. This paper describes a solution to this problem which uses superconducting coils closely integrated with fully saturated iron elements. A 4-T, 30-cm-bore prototype, based on this design principle, was built and tested. The 5 G fringe field is contained within 1 meter of the magnet bore along the z axis. Homogeneity of the raw magnetic field is 10 ppm over 30% of the magnet's diameter after passive shimming. Compared with an unshielded magnet, 20% less superconductor is required to generate the magnetic field. Images and spectra are presented to demonstrate the magnet's viability for magnetic resonance imaging and spectroscopy. PMID- 8419741 TI - Quantitative measurement of velocity at multiple positions using comb excitation and Fourier velocity encoding. AB - An MR imaging technique that simultaneously acquires Fourier velocity encoded data from multiple stations is described. The technique employs a comb excitation rf pulse that excites an arbitrary number of slices. As the Fourier velocity phase encoding gradient pulse is advanced, the phase of each slice is the comb is advanced by a unique amount. This causes the signals from the spins in a particular slice to appear at a position in the phase encoding direction, which is the sum of the spin velocity and an offset arising from the phase increment given to that excitation slice. Acquisition of spin velocity information occurs simultaneously for all slices, permitting the calculation of wave velocities arising from pulsatile flow. These wave velocities can then be used to determine vessel distensibility. PMID- 8419742 TI - In vivo 31P NMR studies of paraplegics' muscles activated by functional electrical stimulation. AB - The bioenergetics of paralyzed muscles of spastic paraplegic patients under functional electrical stimulation (FES) was studied in vivo using 31P NMR. The protocol included rest, 3 min of induced tetanic isometric contraction through surface electrodes and 40 min of recovery. The continuous stimulation, the force recording and the 31P NMR measurements were sampled simultaneously inside the whole body imager. Normal values were found for the phosphorous metabolite ratios at rest. During contraction, prominent changes were detected including: a) accumulation of inorganic phosphate (Pi) accompanied by an unusually strong signal in the phosphomonoester (PME) region, b) phosphocreatine (PCr) decline, and c) a decrease in the intracellular pH. In the following recovery period the physiological state of the muscle was monitored and quantitated by 31P NMR. No metabolic and mechanical irreversible damage was detected in the paraplegics' muscles activated by FES under our experimental conditions. PMID- 8419743 TI - A new approach to study cardiac motion: the optical flow of cine MR images. AB - We present a novel approach to study the intricate motion of the heart using the optical flow technique applied to cine MR images. The method uses image brightness variations between consecutive frames to compute in-plane displacements or velocities of moving image features. The dense velocity field thereby obtained throughout the myocardial wall allows to not only characterize segmental left ventricular wall motion but also to resolve "fine" motion such as intramural differences in displacements and therefore in thickening. Other subtle features of systole like the descent of the base towards the apex or the counterclockwise rotation of the apex with respect to the base can also be detected by the algorithm. Contrary to other techniques proposed earlier, this noninvasive method presents the additional advantages of not requiring any special pulse sequence nor well defined endocardial and epicardial outlines. PMID- 8419744 TI - NMR measurement of 39K detectability and relaxation constants in rat tissue. AB - Differences in the NMR detectability of 39K in various excised rat tissues (liver, brain, kidney, muscle, and testes) have been observed. The lowest NMR detectability occurs for liver (61 +/- 3% of potassium as measured by flame photometry) and highest for erythrocytes (100 +/- 7%). These differences in detectability correlate with differences in the measured 39K NMR relaxation constants in the same tissues. 39K detectabilities were also found to correlate inversely with the mitochondrial content of the tissues. Mitochondria prepared from liver showed greatly reduced 39K NMR detectability when compared with the tissue from which it was derived, 31.6 +/- 9% of potassium measured by flame photometry compared to 61 +/- 3%. The detectability of potassium in mitochondria was too low to enable the measurement of relaxation constants. This study indicates that differences in tissue structure, particularly mitochondrial content are important in determining 39K detectability and measured relaxation rates. PMID- 8419745 TI - Determination of saturation factors in 31P NMR spectra of the developing human brain. AB - In order to assess the influence of longitudinal relaxation on previously reported variations in 31P NMR signals during brain development, we used an accelerated two-point technique to determine T1 at 2.35 Tesla in 8 min. Comparison between 10 normal neonates (age range 37-46 weeks postconception) and 10 healthy infants (age range 80-157 weeks postconception) indicated that T1 does not vary substantially during the first year of life, except in the phosphomonoester (PME) region of the spectra. T1 of total PME decreases with age which we could explain by its variable multicomponent nature: The signal from (unresolved) components at the downfield shoulder of PME (attributed mostly to phosphorylethanolamine at 6.72 ppm) with a T1 of at least 6.4 s decreases with age relative to contributions from other phosphomonoester compounds resonating predominantly at the upfield side of the peak (approximately 6.3 ppm), with T1 below 2.9 s. Because the T1 heterogeneity of PME may depend on its relative composition, quantitative 31P NMR spectroscopy may require an assessment of the influence of longitudinal relaxation on the signal amplitudes in each measurement. PMID- 8419746 TI - Distortions from curved flow in magnetic resonance imaging. AB - We present an analysis of how vessel curvature can create distortions in magnetic resonance images of flowing blood. Steady flow in curved vessels produces distortions of the vessel shape and intensity variations in the image due to motion during the interval between phase encoding or slice selection and the echo center. Even with steady flow, vessel curvature produces motion moments higher than velocity (acceleration, etc.), but use of a first order oblique flow compensated phase encoding gradient waveform reduced the distortion in the image. Numerical calculations of image distortions based on simple flow models are in good agreement with experimental results in a phantom. PMID- 8419747 TI - Hypoxia and metabolic acidosis in the isolated heart: evidence for synergistic injury. AB - Although hypoxia and metabolic acidosis have both been shown to impair cardiac function, some workers have suggested that acidosis during a period of hypoxia will actually accelerate physiologic recovery from this insult. To address the interactions of metabolic acidosis and hypoxia further, isolated isovolumic rat hearts were exposed to normal perfusion conditions for 30 min to establish baseline conditions, then either continued normal conditions, metabolic acidosis, hypoxia, or combined acidosis and hypoxia for 30 min and subsequently reperfused under normal perfusion conditions for an additional 30 min. We observed that acidosis + hypoxia impaired recovery of cardiac contraction more than acidosis or hypoxia alone following experimental perfusion. The combination of acidosis and and hypoxia also impaired cardiac energy metabolism more than acidosis or hypoxia alone as assessed by increases in tissue inorganic phosphate during experimental perfusion as well as during reperfusion. These data suggest that during hypoxia, acidosis appears to primarily impair cardiac energy production as we have previously observed in the normoxic isolated rat heart. Therefore, in the intact beating heart, acidosis may not protect from hypoxic injury as has been suggested in simpler systems but may not protect from hypoxic injury as has been suggested in simpler systems but rather may exacerbate at. PMID- 8419748 TI - MR microscopy of the rat lung using projection reconstruction. AB - Projection reconstruction has been implemented with self-refocused selection pulses on a small bore, 2.0 T MR microscope, to allow imaging of lung parenchyma. Scan synchronous ventilation and cardiac gating have been integrated with the sequence to minimize motion artifacts. A systematic survey of the pulse sequence parameters has been undertaken in conjunction with the biological gating parameters to optimize resolution and signal-to-noise (SNR). The resulting projection images with effective echo time of < 300 microseconds allow definition of lung parenchyma with an SNR improvement of approximately 15 x over a more conventional 2DFT short echo gradient sequence. PMID- 8419749 TI - HIV-related respiratory disease. AB - The lungs are a primary target for the opportunistic infections and malignancies affecting those with HIV infection. In the patient whose HIV infection is undiagnosed, PCP is the commonest clue to its presence. Early diagnosis prevents morbidity and mortality. Less commonly, interstitial lung disease and tuberculosis, often "primary" or clinically atypical, will be the clue to underlying HIV infection. Other pulmonary complications are usually a late manifestation of HIV infection, which has usually (but not always) already been diagnosed. PMID- 8419750 TI - HIV-related oral disease. AB - Oral lesions are frequently an early sign of HIV infection. However, without careful examination their significance can be missed, as they are often painless and discrete and may be the only sign of HIV infection. PMID- 8419751 TI - HIV-related musculoskeletal disease. AB - Among the less well-known manifestations of HIV infection are the musculoskeletal disorders. Since most of these occur in patients who are otherwise well, they represent an important opportunity for the detection of unsuspected HIV infection. PMID- 8419752 TI - Cryptosporidial diarrhoea in South Australia. An exploratory case-control study of risk factors for transmission. AB - OBJECTIVE: To identify risk factors for transmission of cryptosporidiosis in South Australia. DESIGN: Case-control study of 51 cases of laboratory confirmed cryptosporidiosis and 51 age and sex matched controls. SETTING: Subjects from greater Adelaide, with cases notified by local pathology laboratories to the Communicable Disease Control Unit, South Australian Health Commission, during the summer of 1990/1991. PARTICIPANTS: One in 10 cases was selected systematically from 479 laboratory notifications, and permission was obtained from the treating physicians to contact the patients. Subjects nominated age and sex matched controls living in the same area. METHODS: By means of a structured questionnaire, participants were asked by telephone about exposure to possible risk factors in the two weeks preceding the illness/interview. The risk factors included those most commonly cited in the literature as resulting in zoonotic, waterborne and person-to-person infection. The number and percentage of cases and controls exposed was recorded for each risk factor. The probability of having been exposed to selected risk factors was compared between cases and controls by the exact test for matched pairs. RESULTS: The proportion of cases and controls exposed was similar for all risk factors except water sources. Controls were more likely to have consumed only rain water than were cases (P < 0.005). Cases tended more than controls to have consumed only spring water (P = 0.06) or only mains water (P = 0.09). CONCLUSIONS: The consumption of spring water or mains water contaminated with cryptosporidial oocysts may be the mode of transmission of cryptosporidiosis in South Australia. The advent of specific methods for detecting Cryptosporidium sp. in water will allow this hypothesis to be tested. PMID- 8419753 TI - Clinical rheumatology training of Australian medical students. A national survey of 1991 graduates. AB - OBJECTIVE: To describe Australian medical graduates' knowledge, experiences and practical training in rheumatology and their attitudes towards rehabilitation and disability. DESIGN: Cross-sectional survey of all interns at randomly selected hospitals in each State. PARTICIPANTS: 382 Australian interns at 12 hospitals surveyed in the first week of their 1991 internship. RESULTS: New interns demonstrated little experience with soft tissue rheumatism, with only 45% reporting they had examined a patient with bursitis and 22% one with epicondylitis. There was considerable dissatisfaction with the teaching of assessment of low back pain, regardless of the amount of formal rheumatology teaching the graduates had experienced, with only 22% rating it as good or excellent. There was little evidence that students are exposed to the social dimensions of chronic illness; only 32% of students reported that they had been shown how to assess a patient's psychological adjustment to illness. Only 22% felt competent at assessing disability and handicap and less than half of the graduates studied had ever attended a clinic where there was a physiotherapist. Graduates who had never been attached to either a rheumatology ward or an outpatients clinic (17%) were less likely to have examined a patient with gout (P < 0.001), osteoarthritis (P < 0.01), or chronic low back pain (P < 0.05), and were more likely to report dissatisfaction with training in rheumatology. CONCLUSION: This survey suggests that there are significant problems in the training of medical students in musculoskeletal disorders, particularly in relation to the assessment of disability and the appreciation of psychosocial factors. PMID- 8419754 TI - Donation and retrieval of cadaveric organs in Australia. Accepting the challenge. AB - Most transplant programs in this country are significantly restricted by a chronic shortage of donor organs. This article examines the sources of transplantable organs, the concept of brain death, reasons why potential donors may be missed in hospitals, the often perceived difficulty in approaching next-of kin, and the medical requirements and logistics of organ retrieval. The lives of many people are affected by organ donation and transplantation. For those receiving a transplant it can be a gift of incalculable value; for those who agree to donation it may represent one positive outcome from an otherwise tragic situation. These unique circumstances provoke a wide range of emotions and reactions. Health care professionals need to act responsibly towards both donor families and potential transplant recipients. Their involvement in the process of organ donation is an important part of their role in the care of critically ill or injured patients. PMID- 8419755 TI - Lignocaine metabolism and liver function testing in primary graft failure following orthotopic liver transplantation. AB - OBJECTIVE: To report the use of lignocaine and the measurement of its metabolite, monoethylglycinexylidide (MEGX), as a dynamic test of liver function. CLINICAL FEATURES: The conversion of lignocaine to MEGX has been used serially as a test of liver function in a liver transplant recipient for whom retransplantation was necessary because of primary non-function (PNF) of the initial graft. MEGX concentrations were markedly depressed with individual episodes of PNF, cardiac failure and rejection in this patient. CONCLUSION: The test provided useful additional supportive information in assessment of the patient and management of intercurrent problems following liver transplantation. PMID- 8419756 TI - Paradoxical embolus. AB - OBJECTIVE: To present two cases of paradoxical embolism. CLINICAL FEATURES: A 78 year-old woman presented to a hospital emergency department, seven weeks after a right total hip replacement, with chest tightness and sudden-onset left arm and leg pain and paraesthesia. Scanning showed multiple pulmonary emboli, right iliofemoral vein thrombosis, left popliteal and axillary artery embolism, a patent foramen ovale and a right to left shunt. A 41-year-old woman, with a 10 year history of Sjogren's syndrome, presented with sudden-onset left foot pain. Scanning showed emboli in the popliteal and profunda femoris arteries of the left leg, thrombosis involving the right deep calf veins, popliteal vein and superficial femoral vein, and a patent foramen ovale. Based on these critical findings, a diagnosis of paradoxical embolism was made in each case. INTERVENTION AND OUTCOME: Both patients required arterial embolectomy. After commencement of anticoagulant therapy there were no further episodes of embolism. CONCLUSIONS: Paradoxical embolism is an important syndrome which requires a high degree of clinical suspicion as well as several specific investigations for prompt diagnosis. PMID- 8419757 TI - Rescue on the "rattler". Rescue and evacuation by railway ambulances plus railroad-centred health services. PMID- 8419758 TI - Cheap thrills. PMID- 8419759 TI - Trends in alcohol, smoking and drug use among employees. PMID- 8419760 TI - A hand burn from unmarked hydrofluoric acid. PMID- 8419761 TI - CME--the government, universities and colleges. PMID- 8419762 TI - CME--the government, universities and colleges. PMID- 8419763 TI - Breaking bad news. PMID- 8419764 TI - Atrial fibrillation. PMID- 8419765 TI - Whiplash in Australia: illness or injury. PMID- 8419766 TI - Whiplash in Australia: illness or injury. PMID- 8419767 TI - Chronic sport-induced hypohydration. PMID- 8419768 TI - Rural mental health. PMID- 8419769 TI - Ethics committees: is the tail wagging the dog? PMID- 8419770 TI - The day care clinic for dermatology. PMID- 8419771 TI - Titration of immunotherapy. PMID- 8419772 TI - Venesection as protection against myocardial infarction. PMID- 8419773 TI - Hepatotoxicity due to erythromycin ethylsuccinate. PMID- 8419774 TI - Gastric polyposis: onset during long-term therapy with omeprazole. PMID- 8419775 TI - Recycle your journal and other information resources. PMID- 8419776 TI - The excimer laser in Australia. PMID- 8419777 TI - Detection of jeopardised viable myocardium in patients with coronary artery disease. PMID- 8419778 TI - PET joins SPECT in Australian nuclear medicine. PMID- 8419779 TI - New directions for acute stroke therapy. PMID- 8419780 TI - Ascertaining the true incidence of stroke: experience from the Perth Community Stroke Study, 1989-1990. AB - OBJECTIVE: To determine the age and sex specific incidence, and case fatality of stroke in Perth, Western Australia. DESIGN AND SETTING: A population-based descriptive epidemiological study. SUBJECTS: All residents of a geographically defined segment of the Perth metropolitan area (population 138,708) who had a stroke or transient ischaemic attack between 20 February 1989 and 19 August 1990, inclusive. MAIN OUTCOME MEASURES: Definite acute "first-ever-in-a-lifetime" (first-ever) and recurrent stroke classified according to standard definitions and criteria. RESULTS: During the 18-month study period, 536 stroke events occurred among 492 patients, 69% of which were first-ever strokes. The crude annual event rate for all strokes was 258 (95% confidence interval 231-285) per 100,000, and the overall case fatality at 28 days was 24% (95% CI, 20%-28%). The crude annual incidence for first-ever strokes was 178 (95% CI, 156-200) per 100,000; 189 (95% CI, 157-221) per 100,000 in males and 166 (95% CI, 136-196) per 100,000 in females. The corresponding rates, age-adjusted to the "world" population, were 132 (95% CI, 109-155) for males and 77 (95% CI, 60-94) for females. CONCLUSIONS: In contrast to mortality rates for ischaemic heart disease, the incidence of stroke in Australia appears little different from that for several other Western countries. For both males and females the incidence of stroke rises exponentially with increasing age. Although the sex-dependent difference in the risk of stroke is greatest in middle age, males are at greater risk of stroke even among the most elderly. To determine the incidence of stroke accurately, population-based studies of stroke need exhaustive and overlapping sources of case ascertainment. If only cases admitted to hospital had been used, we would have underestimated the rate of stroke among the most elderly by almost 40%. We estimate that approximately 37,000 people, about 50% of whom are over the age of 75, suffer a stroke each year in Australia. PMID- 8419781 TI - Determining the incidence of different subtypes of stroke: results from the Perth Community Stroke Study, 1989-1990. AB - OBJECTIVE: To determine the incidence and case fatality of seven distinct subtypes of stroke in Perth, Western Australia. DESIGN AND SETTING: A population based descriptive epidemiological study. SUBJECTS: All residents of a geographically defined segment of the Perth metropolitan area (estimated population 138,708 persons) who had a stroke or transient ischaemic attack between 20 February 1989 and 19 August 1990, inclusive. MAIN OUTCOME MEASURES: The following subtypes of stroke were classified according to standard clinical, radiological and pathological criteria: types of cerebral infarction, namely, large artery (thrombotic) occlusive infarction (LAOI), cerebral embolic infarction (EMBI), lacunar infarction (LACI) and boundary zone infarction (BZI); primary intracerebral haemorrhage (PICH); subarachnoid haemorrhage (SAH); and stroke of undetermined cause. RESULTS: Over the 18-month study period 536 stroke events were registered, of which 86% (95% confidence interval, 83%-89%) had a defined "pathological" diagnosis on the basis of computed tomographic scanning, magnetic resonance imaging or necropsy. Cerebral infarction accounted for 71% of cases (95% CI, 68%-75%), PICH 11% (95% CI, 9%-14%) and SAH 4% (95% CI, 2%-5%). The 382 cases of cerebral infarction included LAOI (in approximately 71%), EMBI (15%), LACI (10%) and BZI (5%). While the incidence of all subtypes of stroke increased with age, there were age and sex differences in their proportional frequency, management and prognosis: patients with PICH, SAH and EMBI were more likely to be admitted to hospital, and these conditions carried the highest early case fatality. Over all, the 28-day case fatality was 24% (95% CI, 20%-28%), but varied from 0 for LACI and BZI, to 37% (95% CI, 15%-59%) for SAH and 35% (CI, 23% 47%) for PICH. CONCLUSIONS: In this study, we found considerable differences in incidence rates, the effect of age and sex on incidence rates, and prognosis for the different subtypes of stroke. Hospital-based studies are likely to be selectively biased by emphasising strokes that are severe and require admission to hospital. These data have important implications in the design and evaluation of clinical trials of therapy for stroke. PMID- 8419782 TI - Injectable collagen for type 3 female stress incontinence: the first 50 Australian patients. AB - OBJECTIVE: To assess the safety, efficacy and durability of injectable glutaraldehyde cross-linked (GAX) collagen in the treatment of type 3 female stress incontinence. DESIGN AND SETTING: A prospective study of incontinent women attending a private practice for clinical and urodynamic assessment. PATIENTS: Fifty women who had had an average of 1.8 previous operations for stress urinary incontinence took part in the study. Each had been diagnosed as having type 3 genuine stress incontinence (poor or nonfunctioning urethral sphincter mechanism in the presence of a bladder neck which is well supported, usually by scar tissue from previous surgery). Subject to a negative skin test for allergy, each patient underwent the implantation of GAX collagen (Contigen), a biocompatible product causing no foreign body reaction. INTERVENTIONS: The implant was performed by a short day-only procedure involving cystoscopically controlled periurethral (71%) or transurethral (29%) injection of Contigen. The aim was to achieve closure of the urethra at the bladder neck and increase resistance to urine loss. Top-up injections were used as required. The follow-up period ranged from one to 21 months (mean, 11 months). RESULTS: Of these first 50 female patients, 41 (82%) were successfully treated. Twenty-one (42%) were no longer incontinent; 20 (40%) desired no further treatment because their condition was improved; seven (14%) did not respond to treatment and two patients were awaiting top-up injections. The average number of injections given was 1.9 and the average volume injected was 14.4 mL. There were no allergic reactions or infections. Side effects were temporary and of a minor nature. CONCLUSIONS: Injectable GAX collagen appears to be a safe and effective treatment for type 3 stress urinary incontinence. Its durability awaits further follow-up. PMID- 8419783 TI - Experience with nimodipine in aneurysmal subarachnoid haemorrhage. AB - OBJECTIVES: To assess the efficacy of nimodipine in preventing delayed ischaemic deficit in aneurysmal subarachnoid haemorrhage. DESIGN: A continuous prospective audit of all patients with aneurysmal subarachnoid haemorrhage admitted to the joint neurosurgery units of Prince Henry's and Alfred hospitals, Melbourne. Patients were divided into two groups--135 in the pre-nimodipine group during 1986 to 1989, and 73 in the nimodipine group during 1989 and 1990. MAIN OUTCOME MEASURES: Outcome was measured according to the Glasgow outcome scale and the incidence of delayed ischaemic deficit was recorded. RESULTS: A substantial reduction in the overall incidence of poor outcome was observed, from 37% of patients in the non-nimodipine group, to 20% in the nimodipine group (P = 0.022). Delayed ischaemic deficit occurred in 41% and 21% (P = 0.005), and poor outcome due to delayed ischaemic deficit occurred in 18% and 8% (P = 0.09) respectively. CONCLUSIONS: In our experience, nimodipine appears to have substantially reduced the incidence of delayed ischaemic deficits in patients with aneurysmal subarachnoid haemorrhage, with a resultant improvement in overall patient outcome. PMID- 8419784 TI - Extracorporeal shock wave lithotripsy combined with litholytic therapy in the treatment of patients with symptomatic gallstones--the Melbourne experience. AB - OBJECTIVE: To investigate the role of extracorporeal shock wave lithotripsy using the Dornier MPL9000 lithotripter and adjuvant litholytic therapy in the treatment of symptomatic gallbladder stones. PATIENTS AND METHODS: Between August 1989 and March 1991, 399 patients had their one to three gallbladder stones fragmented by the Dornier MPL9000 lithotripter. Chenodeoxycholic acid alone was used as adjuvant litholytic therapy in the majority. A minority received a combination of chenodeoxycholic acid and ursodeoxycholic acid or ursodeoxycholic acid alone. Patients who died, had cholecystectomies or failed to complete the treatment program were excluded from analysis, leaving a cohort of 287 patients with a follow-up of at least 12 months. This cohort comprised 173 patients with single small stones (20 mm or less in diameter), 32 patients with single large stones (21 mm to 30 mm in diameter) and 82 patients with two to three stones. OUTCOME MEASURES: Patients were followed up by repeated ultrasound examination to monitor the disappearance of fragments from the gallbladder. Stone-free rates, recurrences and complications of treatment were determined. RESULTS: The stone free rate 12 months after treatment was 37.6% for patients with a single small stone, 3.1% for patients with a single large stone and 18.3% for patients with two to three stones. Of 70 patients with a single small stone who had become stone free at some time during the 12 months after treatment, five (7.1%) experienced recurrence, as did one of the 16 patients (6.9%) with two to three stones. Some 179 patients (44.9%) experienced biliary colic after lithotripsy. Most attacks were mild. Eleven patients (2.8%) developed cholecystitis and nine (2.3%) became jaundiced. Five patients (1.3%) suffered from pancreatitis, of whom one died from severe necrotising pancreatitis. Treatment mortality was 0.25%. Cholecystectomy was needed in 44 patients (11.9%). CONCLUSIONS: Only about 15% 20% of all patients with symptomatic gallbladder stones are suitable for lithotripsy. In this study, only about 28% were stone free after 12 months. As the gallbladder is not removed, stones may re-form. Laparoscopic cholecystectomy and open cholecystectomy by comparison will produce a "stone-free state" in 100% of patients, no matter how many stones are present in the gallbladder, their size, or whether the gallbladder is non-functioning. Consequently, lithotripsy and litholytic therapy are now reserved for those few patients who are unable to tolerate general anaesthesia and cholecystectomy and those who refuse surgery. Even in centres showing the most favourable results, lithotripsy and litholytic therapy will have at best a minor role to play in the overall management of symptomatic gallbladder stones. PMID- 8419785 TI - Prediction of pregnancy-induced hypertension by means of the urinary calcium:creatinine ratio. AB - OBJECTIVE: To investigate the importance of the urinary calcium:creatinine ratio as a prognostic marker for pregnancy-induced hypertension. DESIGN: A prospective study which measured the urinary calcium:creatinine ratio at 20-30 weeks' gestation. Patients' medical records were examined, blind to all urinary assay results, to determine the development of pre-eclampsia or gestational hypertension. SETTING AND SUBJECTS: A first-morning urine sample was collected from 456 normotensive pregnant women, at 20-30 weeks' gestation, attending a hospital maternity outpatients' clinic for routine antenatal care. RESULTS: The mean urinary calcium:creatinine ratio for women (n = 392) with a normotensive outcome of pregnancy, 0.52 (SD 0.32), was not significantly different from the ratios in those women who developed pre-eclampsia (n = 16), 0.49 (SD 0.32) or gestational hypertension (n = 48), 0.57 (SD 0.41). Significant risk factors for development of gestational hypertension, as estimated by logistic regression, were mean arterial blood pressure greater than 87.6 mmHg, Caucasian race, non smoking and nulliparity. CONCLUSIONS: Within the cohort studied, the calcium:creatinine ratio measured in asymptomatic women at 20-30 weeks' gestation was an unsatisfactory prognostic marker for development of pregnancy-induced hypertension. The major interpopulation, as well as intrapopulation, variation in calcium levels suggests that previous reported findings may not be readily generalised. PMID- 8419786 TI - New advertisements for an old antihistamine. PMID- 8419787 TI - Coccidioidomycosis--United States, 1991-1992. AB - During 1991, reported cases of coccidioidomycosis (i.e., valley fever) in California increased more than three-fold over the annual number of cases reported since 1986; during 1992, the number of reported cases increased 10-fold. Coccidioidomycosis, a fungal disease caused by Coccidioides immitis, is endemic in certain parts of Arizona, California, Nevada, New Mexico, Texas, and Utah. Sporadic cases occur each year in parts of the United States in which the disease is not endemic and may present diagnostic difficulties and laboratory hazards because health-care workers may be unfamiliar with coccidioidomycosis. Recent increases in California and reports of isolated cases in areas without endemic disease suggest that physicians and laboratory personnel should be alert to the possible role of C. immitis. This report summarizes the occurrence of coccidioidomycosis in California during 1991 and 1992 and highlights three cases that occurred in areas in which the disease is not endemic. PMID- 8419788 TI - Gang-related outbreak of penicillinase-producing Neisseria gonorrhoeae and other sexually transmitted diseases--Colorado Springs, Colorado, 1989-1991. AB - In April 1990, the El Paso County (Colorado) Health Department (EPCHD) recognized an outbreak of penicillinase-producing Neisseria gonorrhoeae (PPNG) and other sexually transmitted diseases (STDs) occurring in Colorado Springs (1990 census population: 397,014), Colorado. An investigation by the EPCHD and the Colorado Department of Health eventually identified 56 cases of PPNG from December 1989 through March 1991 (Figure 1). The outbreak revealed a previously unidentified core group of persons with STDs in Colorado Springs. This report summarizes traditional and innovative measures used to investigate and manage the outbreak, and describes this core group and its role in STD transmission in Colorado Springs. PMID- 8419789 TI - Worksite health promotion--New Hampshire, 1992. AB - Because a high proportion (85%) of the U.S. adult population is employed, the worksite setting offers immense potential for health-promotion efforts (1). Successful worksite health-promotion programs have targeted nutrition, cholesterol reduction, and cancer prevention (2-4). As part of an effort to strengthen such programs in New Hampshire, the Division of Public Health Services (DPHS), New Hampshire State Department of Health and Human Services, in collaboration with the University of New Hampshire and CDC, conducted a statewide survey of worksites from March through July 1992 to characterize employee health services. This report summarizes findings on the proportion of worksites that offered health-promotion activities. PMID- 8419790 TI - Preliminary data: exposure of persons aged > or = 4 years to tobacco smoke- United States, 1988-1991. AB - The recent report of the U.S. Environmental Protection Agency on the respiratory health effects of passive smoking (1) and the known adverse effects of active smoking emphasize the need to quantify the exposure of the U.S. population to tobacco smoke. Measurements of cotinine (a nicotine metabolite) in serum, urine, and saliva have been used effectively to quantify exposure to tobacco smoke (2 10). As part of the Third National Health and Nutrition Examination Survey (NHANES III), CDC's National Center for Environmental Health and National Center for Health Statistics is measuring serum levels of cotinine to assess exposure to tobacco smoke by persons in the United States aged > or = 4 years. This report presents preliminary findings on the first 800 persons in this survey of tobacco smoke exposure. PMID- 8419791 TI - Ceftriaxone-associated biliary complications of treatment of suspected disseminated Lyme disease--New Jersey, 1990-1992. AB - Lyme disease (LD) is endemic in Monmouth and Ocean counties, New Jersey (1). In June 1992, CDC and the New Jersey Department of Health (NJDOH) conducted a telephone survey in both counties of 65 schoolchildren who required home instruction because of suspected LD to determine the public health impact of the disease. Most children had received prolonged and repeated courses of oral antimicrobials and/or home intravenous infusion of antimicrobials; 79% had been hospitalized for treatment of suspected LD or management of treatment complications, most notably drug-induced symptoms of gallbladder disease occurring in patients receiving ceftriaxone (Rocephin), and bloodstream infections associated with intravenous catheters. To determine the characteristics of and treatment complications for patients hospitalized for treatment of LD, a computerized search of hospital discharge data in New Jersey was performed; nearly 30% of all hospitalizations for LD during 1990-1991 were at a regional hospital serving Monmouth and Ocean counties. This report presents findings of an analysis of patients admitted to that hospital for treatment of LD. PMID- 8419792 TI - The possible role of iron in the etiopathology of Parkinson's disease. AB - The identification of 6-hydroxydopamine (6-OHDA) and N-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) as dopaminergic neurotoxins that can induce parkinsonism in humans and animals has contributed to a better understanding of Parkinson's disease (PD). Although the involvement of similar neurotoxins has been implicated in PD, the etiology of the disease remains obscure. However, the recently described pathology of PD supports the view for a state of oxidative stress in the substantia nigra (SN), resulting as a consequence of the selective accumulation of iron in SN zona compacta and within the melanized dopamine neurons. Whether iron is directly involved cannot be ascertained. Nevertheless, the biochemical changes due to oxidative stress resulting from tissue iron overload (siderosis) are similar to those now being identified in parkinsonian SN. These include the reduction of mitochondrial electron transport, complex I and III activities, glutathione peroxidase activity, glutathione (GSH) ascorbate, calcium-binding protein, and superoxide dismutase and increase of basal lipid peroxidation and deposition of iron. The participation of iron-induced oxygen free radicals in the process of nigrostriatal dopamine neuron degeneration is strengthened by recent studies in which the neurotoxicity of 6-OHDA has been linked to the release of iron from its binding sites in ferritin. This is further supported by experiments with the prototype iron chelator, desferrioxamine (Desferal), a free-radical inhibitor, which protects against 6-OHDA-induced lesions in the rat. Indeed, intranigral iron injection in rats produces a selective lesioning of dopamine neurons, resulting in a behavioral and biochemical parkinsonism. PMID- 8419793 TI - Movement disorders with cerebral toxoplasmosis and AIDS. AB - Movement disorders occur in some patients with cerebral toxoplasmosis with HIV-1 infection. Such movement disorders have not been described in patients with cerebral toxoplasmosis without HIV-1 infection. This report discusses their diagnostic features, aspects of management, and possible mechanisms underlying the pathogenesis of the movement disorders. PMID- 8419795 TI - Torticollis after electrocution. PMID- 8419794 TI - Akinetic-rigid syndrome in a 13-year-old girl with HIV-related progressive multifocal leukoencephalopathy. AB - A 13-year-old HIV-infected girl presented with a rapidly progressive akinetic rigid syndrome. Cranial magnetic resonance imaging revealed bilateral centrum semiovale lesions extending into the basal ganglia. Detailed histopathological study of a biopsy specimen from the involved regions resulted in a diagnosis of progressive multifocal leukoencephalopathy (PML). This case illustrates that an akinetic-rigid syndrome may be a rare presentation of PML. HIV-related PML may have unusual radiological features. Brain biopsy may be necessary to confirm PML in cases associated with atypical clinical and radiological presentations. PMID- 8419796 TI - Hormonal influences on spasmodic torticollis: a case report. PMID- 8419797 TI - Generalized chorea induced by nonketotic hyperglycemia. PMID- 8419798 TI - Bifrontal glioma presenting as a gross movement disorder. PMID- 8419799 TI - Alcohol sensitive dystonia. PMID- 8419800 TI - Origin of chorea-ballism and multisystem degeneration: pathophysiological implications. PMID- 8419801 TI - Drug-induced parkinsonism: a growing list. PMID- 8419802 TI - Transient dyskinesia induced by clebopride. PMID- 8419803 TI - Reaction time responses in parkinsonian and hemiparkinsonian patients. AB - Twenty-one normal subjects, 32 bilateral parkinsonian patients (BPs) and 29 hemiparkinsonian patients (HP) were submitted to separate or sequentially associated motor tasks that included simple reaction times (RT), choice RTs, directional RTs, and movement RTs. The results showed that simple RTs, directional RTs, and movement times (MT) were slower in BPs as compared to normal subjects; for choice RTs there was no difference. Response patterns were similar in normal controls and BPs. In both groups RTs became more prolonged as sequentially programmed operations increased. If movement occurred at the end of the sequence, they prolonged the RTs of the preceding operations, but MTs per se did not vary. In HPs the same results were observed on the "bad" hand side versus normal controls and versus the healthy side, but a significant statistical level was reached mainly when the "bad" hand was the right one. PMID- 8419804 TI - Phenytoin and dyskinesias: a report of two cases and review of the literature. AB - Dyskinesia is a recognized but uncommon side-effect of treatment with phenytoin. Two additional cases of dyskinesia during treatment with phenytoin are described; both had radiographically documented thalamic infarctions. The reported experience to date with movement disorders induced by phenytoin is reviewed and the clinical features summarized. The available experimental evidence addressing the mechanism underlying this side effect is discussed. PMID- 8419805 TI - The cholinergic system-dependent myoclonus of the baboon Papio papio is a reticular reflex myoclonus. AB - Neurophysiological studies were performed on four Papio papio baboons presenting with nonepileptic myoclonus (a startle response resembling stimulus-sensitive jerk). Investigations of the EEG, back-averaged EEG, and somatosensory evoked potentials revealed the absence of cortical correlates preceding the jerks, and exclusion of cerebral cortex involvement. No long-latency reflexes could be recorded in these animals. The jerks were symmetric when evoked by unilateral stimulation in normal baboons as well as in a split-brain animal. Polymyographic records showed that the first muscle involved during the jerk was the trapezius; other muscles were involved with latencies increasing in both cranial and caudal directions. From these data, nonepileptic myoclonus of baboons can be classified as a reticular reflex myoclonus. The involvement of cranial nerves did not follow the layout of the nuclei in the brainstem, indicating that the jerk is most likely generated as a complete movement. The generating structure is probably under cholinergic control. Finally, the Papio papio baboon, which was already known as a model for cortical myoclonus elicited by intermittent photic stimulation in predisposed animals, can also be considered a model for the study of the reticular reflex myoclonus. PMID- 8419806 TI - Quantitative measurement of cervical range of motion in patients with torticollis treated with botulinum A toxin. AB - Improvement in cervical range of motion in patients with spasmodic torticollis by botulinum A toxin injection is difficult to objectively measure. Recently, a three-dimensional cervical range of motion system (EMROM) that measures primary as well as secondary cervical angles has been developed. This system uses an electromagnetic tracking system for data collection and a personal computer for analysis and graphic display of the data. We have tested the EMROM system and, from our results, believe that it can be used clinically to objectively and accurately measure cervical range of motion in patients who have spasmodic torticollis and who receive botulinum toxin injections. PMID- 8419807 TI - Neuropsychological correlates of brain atrophy in Parkinson's disease: a CT-scan study. AB - We examined the presence of cortical or subcortical brain atrophy (as shown by CT scans) in patients with Parkinson's disease (PD) and assessed whether there were significant correlations between CT measurements and the presence of cognitive deficits. There were three main findings. First, patients with bilateral symptoms of PD showed more severe cortical and subcortical atrophy than age-matched normal controls. Second, the presence of unilateral symptoms of PD was significantly associated with contralateral brain atrophy only in patients with right hemi-PD. Third, there was a significant correlation between neuropsychological deficits and atrophy in specific brain areas. PMID- 8419808 TI - Single-dose L-dopa response in early Parkinson's disease: measurements with optoelectronic recording technique. AB - Twenty-five patients with suspected Parkinson's disease were submitted to optoelectronic movement analysis with the Posturo-Locomotor-Manual (PLM) test before and 60 min after a single dose of L-Dopa. They were then examined clinically for diagnosis. Two patients were excluded due to L-Dopa intolerance. Seventeen of the remaining patients were classified as having Parkinson's disease. The movement time (MT) in the PLM test was increased for all these patients, and they improved their performance after L-Dopa. The degree of improvement was roughly proportional to the pretreatment augmentation of MT in comparison to healthy subjects of the same age. The PLM phase analysis showed a specific disability profile for each individual. Six patients were given diagnoses other than PD. Some improvement was found in one patient with suspected olivopontocerebellar atrophy and one patient with multiple brain injury. Two patients with progressive supranuclear palsy, one with suspected striatonigral degeneration, and one with functional disturbance deteriorated after L-Dopa. In conclusion, truly objective and fully reproducible evaluation of the motor performance before and after a single L-Dopa dose is easily accomplished with computer-assisted modern optoelectronic recording equipment. The technique is a valuable tool for the quantitative measurement of treatment effects and contributes to the differential diagnosis. PMID- 8419809 TI - Effectiveness of piracetam in cortical myoclonus. AB - Twenty-one patients with disabling spontaneous, reflex, or action myoclonus due to various causes, who had shown apparent clinical improvement on introduction of piracetam, entered a placebo-controlled double-blind crossover trial of piracetam (2.4-16.8 g daily). All but one patient had electrophysiological evidence of cortical myoclonus. Patients were randomly allocated to a 14-day course of piracetam followed by identical placebo, or placebo followed by piracetam. Nineteen patients received piracetam/placebo in addition to their routine antimyoclonic treatment (carbamazepine, clonazepam, phenytoin, primidone, sodium valproate, or tryptophan plus isocarboxazid, alone or in combination) and two received piracetam/placebo as monotherapy. All patients were rated at the end of each treatment phase using stimulus sensitivity, motor, writing, functional disability, global assessment, and visual analogue scales. Ten of the 21 patients had to be rescued from the placebo phase of the trial because of a severe and intolerable exacerbation of their myoclonus. No patients required rescue from the piracetam phase of the double-blind trial. When the 21 patients were considered together, there was a significant improvement in motor, writing, functional disability, global assessment, and visual analogue scores during treatment with piracetam compared with placebo. The total rating score also improved significantly with piracetam, by a median of 22%. Piracetam, usually in combination with other antimyoclonic drugs, is a useful treatment for myoclonus of cortical origin. PMID- 8419810 TI - Asymmetric catalepsy after right hemisphere stroke. AB - We describe the appearance of left hemineglect and striking cataleptic posturing, more prominent in left-sided extremities, in a patient without psychiatric illness. Neuroimaging demonstrated a large posterior right hemisphere infarct involving the parietal, occipital, and temporal lobes, the insula, and caudate. Additional movement abnormalities that comprise the full catatonia syndrome were absent, including stereotypy, mannerisms, ambitendency, automatic obedience, mutism, negativism, and echopraxia. Catatonia has been reported to be produced by lesions of diverse etiology affecting the frontal lobe, limbic system, diencephalon, or basal ganglia. In these cases, catalepsy has been manifest only rarely, and motor signs that are present are generally bilateral. This case demonstrates that asymmetric catalepsy can be produced by right hemisphere stroke, and provides partial support for earlier clinical literature relating catalepsy and the parietal lobe. PMID- 8419811 TI - PCR analysis of platelet mtDNA: lack of specific changes in Parkinson's disease. AB - An alteration within the mitochondrial DNA (mtDNA) has been hypothesized to underlie the deficiencies in mitochondrial complex I activity observed in the platelets, striatal muscle, and brain tissue of individuals with Parkinson's disease. Here we utilized the polymerase chain reaction (PCR) to analyze mtDNA obtained from the platelets of nonmedicated patients with early Parkinson's disease (n = 8) and aged-matched controls (n = 6) for the presence of deletion(s) or addition(s) equal to or greater than 50-100 base pairs. Initial attention was focused upon detecting a 4.977 kb deletion previously found in the brains of parkinsonian patients and some aged controls. Indeed, a large deletion of approximately 5.0 kb was observed in the platelet mtDNA from all parkinsonian individuals. However, this defect was also found in all age-matched controls as well as in a group of young healthy subjects (n = 5). In addition, we searched for the presence of smaller changes in platelet mtDNA from parkinsonian patients by PCR analysis of four mtDNA segments that code for seven of the complex I polypeptides. No large deletions or additions were detected within these four regions of mtDNA in any of the disease or age-matched control samples. We conclude that (a) a 4.977 kb deletion is apparently present in a subpopulation of platelet mtDNA from all individuals, and (b) no macrosequence alteration in mtDNA is likely to underlie the deficiency in complex I activity reported in platelet mitochondria from parkinsonian patients. PMID- 8419812 TI - Parkinson's disease mortality and the industrial use of heavy metals in Michigan. AB - Parkinson's disease (PD) mortality rates in Michigan counties for 1986-1988 were calculated with respect to potential heavy metal exposure (iron, zinc, copper, mercury, magnesium, and manganese) from industry based on recent census data. Individuals were counted as a PD death if the diagnosis was listed as an "underlying" or "related" cause of death on the death certificate. Counties with an industry in the paper, chemical, iron, or copper related-industrial categories (ICs) had statistically significantly (p < 0.05) higher PD death rates than counties without these industries. Significant correlations of chemical (rs = 0.22; p = 0.05), paper (rs = 0.22; p = 0.05) and iron (rs = 0.29; p = 0.008) industry densities with PD death rates were also present. Counties were divided into high (> 15/100,000 individuals 45 years old and over) and low (< = 15/100,000) PD death rate counties by cluster analysis. Geographically, counties with high PD mortality were located mainly in the southern half of the lower peninsula and eastern half of the upper peninsula; low PD death rate counties formed two distinct clusters in the western edge of the upper peninsula and the north-central portion of the lower peninsula. Other possible risk factors that may explain the varied distribution of PD death rates in Michigan were examined. Those significantly correlated with PD mortality included population density (rs = 0.31; p = 0.005), farming density (rs = 0.25; p = 0.02), and well water use (rs = -0.24; p = 0.03). These ecologic findings suggest a geographic association between PD mortality and the industrial use of heavy metals. PMID- 8419813 TI - Comparison of somatosensory evoked potentials with striatal glucose consumption measured by positron emission tomography in the early diagnosis of Huntington's disease. AB - Both somatosensory evoked potentials (SEP) and striatal glucose consumption (rCMRGlc) measured by positron emission tomography (PET) have been reported to be abnormal early in the course of Huntington's disease (HD). To compare their diagnostic value, SEP and rCMRGlc were measured in a group of 18 first degree off spring of HD families: 6 had manifest HD with chorea and the remaining 12 individuals were chorea-free subjects at risk for HD. In five patients with manifest disease, both SEP and striatal rCMRGlc were significantly abnormal, defined in SEP as having either a bilaterally absent frontal N30 amplitude or a reduction of the parietal N20/P25 amplitude below 1 microV on at least one side; in PET as exhibiting a reduction of the cerebellar ratio (CR) of both caudate and lentiform rCMRGlc below the 99% confidence limits of these variables determined in 20 normal volunteers. The remaining patient with manifest HD had questionably abnormal SEP and significantly reduced indices of striatal rCMRGlc. The five persons at risk for HD who had normal SEP also had normal striatal rCMRGlc; those three at-risk patients with abnormal SEP also had a reduction of the CR of both caudate and lentiform rCMRGlc. Of the remaining four individuals at risk for HD who had questionably abnormal SEP, three had CR values of striatal rCMRGlc in the normal range and one a reduction of the CR of lentiform rCMRGlc. In at-risk patients, the SEP diagnosis correlated significantly with caudate (r = -0.8; p < 0.002) and lentiform (r = -0.76; p < 0.005) rCMRGlc. These data indicate a parallel deterioration of SEP and striatal rCMRGlc early in the course of HD even before the development of chorea. PMID- 8419814 TI - Rapid serologic testing with immune-complex-dissociated HIV p24 antigen for early detection of HIV infection in neonates. Southern California Pediatric AIDS Consortium. AB - BACKGROUND: Serologic detection of human immunodeficiency virus (HIV) infection in neonates is complicated by the presence of immune complexes, consisting of passively transferred maternal antibodies and HIV antigens. A new, rapid assay has been designed to disrupt these immune complexes in order to permit the detection of a specific HIV antigen. We evaluated the efficacy of this assay in detecting HIV infection in neonates. METHODS: We measured p24 antigen in blood samples from both infected and uninfected children of HIV-infected mothers. The samples were treated with glycine hydrochloride to dissociate the immune complexes, followed by neutralization with TRIS-hydrochloric acid. A commercial HIV p24 antigen assay was then used, with an optical density greater than 0.120 at a wavelength of 450 nm defined as indicating a positive result. RESULTS: Of eight cord-blood samples from neonates with proved HIV infection, five were positive for immune-complex-dissociated p24 antigen. For two other neonates the first postnatal sample, obtained on days 12 and 18, was positive. There was no follow-up sample for the eighth neonate. Of 22 uninfected neonates, 20 were negative on the cord-blood assay. Two neonates had positive cord-blood samples, but the first postnatal sample was negative. Thus, the tests with early postnatal samples identified the HIV-infection status correctly for all 29 children who could be evaluated. In a separate group of 78 children (median age, 188 weeks), the specificity of the test was 100 percent and the sensitivity 81 percent. CONCLUSIONS: The immune-complex-dissociated HIV p24 antigen assay is a rapid, simple serologic test that may be of value in diagnosing HIV infection in neonates born to HIV-infected women. PMID- 8419815 TI - Sympathetic-nerve activity during sleep in normal subjects. AB - BACKGROUND: The early hours of the morning after awakening are associated with an increased frequency of events such as myocardial infarction and ischemic stroke. The triggering mechanisms for these events are not clear. We investigated whether autonomic changes occurring during sleep, particularly rapid-eye-movement (REM) sleep, contribute to the initiation of such events. METHODS: We measured blood pressure, heart rate, and sympathetic-nerve activity (using microneurography, which provides direct measurements of efferent sympathetic-nerve activity related to muscle blood vessels) in eight normal subjects while they were awake and while in the five stages of sleep. RESULTS: The mean (+/- SE) amplitude of bursts of sympathetic-nerve activity and levels of blood pressure and heart rate declined significantly (P < 0.001), from 100 +/- 9 percent, 90 +/- 4 mm Hg, and 64 +/- 2 beats per minute, respectively, during wakefulness to 41 +/- 9 percent, 80 +/- 4 mm Hg, and 59 +/- 2 beats per minute, respectively, during stage 4 of non-REM sleep. Arousal stimuli during stage 2 sleep elicited high-amplitude deflections on the electroencephalogram (called K complexes), which were frequently associated with bursts of sympathetic-nerve activity and transient increases in blood pressure. During REM sleep, sympathetic-nerve activity increased significantly (to 215 +/- 11 percent; P < 0.001) and the blood pressure and heart rate returned to levels similar to those during wakefulness. Momentary restorations of muscle tone during REM sleep (REM twitches) were associated with cessation of sympathetic-nerve discharge and surges in blood pressure. CONCLUSIONS: REM sleep is associated with profound sympathetic activation in normal subjects, possibly linked to changes in muscle tone. The hemodynamic and sympathetic changes during REM sleep could play a part in triggering ischemic events in patients with vascular disease. PMID- 8419816 TI - Effect of ranitidine and amoxicillin plus metronidazole on the eradication of Helicobacter pylori and the recurrence of duodenal ulcer. AB - BACKGROUND: Persistent infection with Helicobacter pylori is associated with the recurrence of duodenal ulcer. Whether the efficacy of bismuth therapy in reducing the rate of recurrence of duodenal ulcer is due to its antimicrobial effects on H. pylori or to a direct protective action on the mucosa is still a matter of debate. METHODS: To study the effect of the eradication of H. pylori on the recurrence of duodenal ulcer, we treated 104 patients with H. pylori infection and recurrent duodenal ulcer with either amoxicillin (750 mg three times daily) plus metronidazole (500 mg three times daily) or identical-appearing placebos, given orally for 12 days. All patients also received ranitidine (300 mg each night) for 6 or 10 weeks. Endoscopy was performed before treatment and periodically during follow-up for up to 12 months after healing. RESULTS: Among the 52 patients given antibiotics, H. pylori was eradicated in 46, as compared with 1 of the 52 given placebo (89 percent vs. 2 percent, P < 0.001). After six weeks, the ulcers were healed in 48 patients given antibiotics and 39 given placebo (92 percent vs. 75 percent, P = 0.011). Side effects, mainly diarrhea, occurred in 15 percent of the patients given antibiotics. Among the patients followed up for 12 months, duodenal ulcers recurred in 4 of 50 patients given antibiotics and 42 of 49 given placebo (8 percent vs. 86 percent, P < 0.001). Ulcers recurred in 1 of 46 patients in whom H. pylori had been eradicated, as compared with 45 of 53 in whom H. pylori persisted (2 percent vs. 85 percent, P < 0.001). CONCLUSIONS: In patients with recurrent duodenal ulcer, eradication of H. pylori by a regimen that does not have any direct action on the mucosa is followed by a marked reduction in the rate of recurrence, suggesting a causal role for H. pylori in recurrent duodenal ulcer. PMID- 8419817 TI - Serum cholesterol in young men and subsequent cardiovascular disease. AB - BACKGROUND: The increased risk of cardiovascular disease associated with higher serum cholesterol levels in middle-aged persons has been clearly established, but there have been few opportunities to examine a potential link between serum cholesterol levels measured in young men and clinically evident premature cardiovascular disease later in life. METHODS: We performed a prospective study of 1017 young men (mean age, 22 years) followed for 27 to 42 years to quantify the risk of cardiovascular disease and total mortality associated with serum cholesterol levels during early adult life. The mean serum cholesterol level at entry was 192 mg per deciliter (5.0 mmol per liter). RESULTS: During a median follow-up of 30.5 years, there were 125 cardiovascular-disease events, 97 of which were due to coronary heart disease. The serum cholesterol level at base line was strongly associated with the incidence of events related to coronary heart disease and cardiovascular disease, as well as to total mortality and mortality due to cardiovascular disease. The risks were similar whether the events occurred before or after the age of 50. In a proportional-hazards analysis adjusted for age, body-mass index (the weight in kilograms divided by the square of the height in meters), the level of physical activity, coffee intake, change in smoking status, and the incidence of diabetes and hypertension during follow up, a difference in the serum cholesterol level at base line of 36 mg per deciliter (0.9 mmol per liter)--the difference between the 25th and 75th percentiles of cholesterol level in the study population at base line--was associated with an increased risk of cardiovascular disease (relative risk, 1.72; 95 percent confidence interval, 1.39 to 2.14), coronary heart disease (relative risk, 2.01; 95 percent confidence interval, 1.59 to 2.53), and mortality due to cardiovascular disease (relative risk, 2.02; 95 percent confidence interval, 1.23 to 3.32). A difference in the base-line serum cholesterol level of 36 mg per deciliter was significantly associated with an increased risk of death before the age of 50 (relative risk, 1.64; 95 percent confidence interval, 1.03 to 2.61), but not with the overall risk of death (relative risk, 1.21; 95 percent confidence interval, 0.93 to 1.58). CONCLUSIONS: These findings indicate a strong association between the serum cholesterol level measured early in adult life in men and cardiovascular disease in midlife. PMID- 8419818 TI - Brief report: reuse of a transplanted heart. PMID- 8419819 TI - Sexual harassment in medical training. AB - BACKGROUND: Sexual harassment has become a national concern and one that is increasingly recognized in the field of medicine. Although there are reports of the sexual harassment of medical trainees, there is little information on the prevalence of this problem and whether it is adequately addressed by training institutions. METHODS: Surveys with descriptions and examples of sexual harassment were mailed to 133 internal medicine residents in a university training program. The residents were asked to report anonymously whether they had encountered sexual harassment during medical school or residency, the frequency and type of harassment, its effect on them, whether they chose to report it to a person in authority, and the factors that influenced this decision. RESULTS: Surveys were returned by 82 residents (response rate, 62 percent), 33 women and 49 men. Twenty-four women (73 percent) and 11 men (22 percent) reported that they had been sexually harassed at least once during their training. The women were more likely than the men to have been physically harassed, and the women's harassers were of higher professional status. Among those harassed, 19 of the women (79 percent) and 5 of the men (45 percent) thought that the experience created a hostile environment or interfered with their performance at work, but only 2 women and no men reported their experiences to an authority. The women cited a lack of confidence that they would be helped as the main reason for not reporting the experience, whereas men most commonly said that they had dealt with the problem without the need for outside assistance. CONCLUSIONS: Many medical trainees encounter what they believe to be sexual harassment during medical school or residency, and this often creates a hostile learning and work environment. Training institutions need to address the adverse effects this may have on medical education and patient care. PMID- 8419820 TI - Weak reasoning: diagnosis by drug reaction. PMID- 8419821 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 5-1993. An 81-year-old man with pain and crepitus in the shoulder. PMID- 8419822 TI - Autonomic modulation of the cardiovascular system during sleep. PMID- 8419823 TI - Treatment of peptic ulcers caused by Helicobacter pylori. PMID- 8419824 TI - Toward a more perfect world--eliminating sexual discrimination in academic medicine. PMID- 8419825 TI - Genetic influence on smoking. PMID- 8419826 TI - Genetic influence on smoking. PMID- 8419827 TI - Seizures after povidone-iodine mediastinal irrigation. PMID- 8419828 TI - The use of albendazole in patients with single lesions enhanced on contrast CT. PMID- 8419829 TI - Electrophysiologically guided antiarrhythmic therapy versus beta-blocker therapy in patients with ventricular tachyarrhythmias. PMID- 8419830 TI - Air-bag-associated rupture of the right atrium. PMID- 8419831 TI - Creutzfeldt-Jakob disease infectivity of growth hormone derived from human pituitary glands. PMID- 8419832 TI - Voracious shredder syndrome. PMID- 8419834 TI - [Lectori salutem?]. PMID- 8419833 TI - The American health care system. The End Stage Renal Disease Program. PMID- 8419835 TI - [Study methods to determine neurological prognosis and therapeutic intervention in full-term neonates with asphyxia]. PMID- 8419836 TI - [Controversies in preventive health care. I. Criteria for the effectiveness of preventive programs]. PMID- 8419837 TI - [Peer review: is one-eye king?]. PMID- 8419838 TI - [Survey of the number of electroencephalograms prepared in The Netherlands in 1975-1989; noticeable decrease in spite of substantially increased number of neurologic patients]. AB - On the basis of data supplied by 89 Dutch neurological/clinical neurophysiological practices it is demonstrated that the number of electroencephalograms (EEGs) prepared in the Netherlands has gradually decreased between 1975 and 1989. In 57 clinics which supplied data on all these years, the decrease was found to amount to about 19%. This significant decrease occurred in spite of a marked increase of the numbers of new patients in the neurological outpatient clinic (about 60%, data from 42 practices) and neurological admissions (about 40%, data from 54 practices). There is no reason to assume that this development has been essentially different in the non-responding practices (n = 59). The findings of this study refute the statement made in an article published in this journal in 1989 that the number of EEGs had increased in spite of the introduction of the CT scanner. This erroneous conclusion, which led to the suggestion of overconsumption, proved to be due in the first place to obviously incorrect figures supplied by the Medical Insurance Board. In addition, the increase of the number of neurological patients had not been taken into account. PMID- 8419839 TI - [Perinatal asphyxia and postpartum resuscitation: always beginning, but when to stop?]. PMID- 8419840 TI - [A Dutch family with hereditary joint symptoms; multiple epiphyseal dysplasia]. AB - Multiple epiphyseal dysplasia is a rare disease with autosomal dominant heredity which affects the epiphyseal ends of long bones; it leads to severe destruction of the joints. A large Dutch family with multiple epiphyseal dysplasia is described with manifestations of clinical disease predominantly in the lower extremities. PMID- 8419841 TI - [Value of an electroencephalogram in a patient with transient neurological symptoms]. AB - A seven year old boy had several episodes of dysfunction of the left hemisphere. The only sign on physical examination was a very slight facial asymmetry on the right side. The CT scan with and without contrast enhancement was normal. Because of a marked asymmetry of the EEG, duplex scan examination of the carotid arteries was carried out; it showed a flow disturbance of the left carotid artery. Angiography showed a narrowing of the supraclinoidal portion of the internal carotid artery which was considered to be due to angiitis. As the boy was known to have bronchitis and eczema and the blood tests showed a marked increase of eosinophil cells, he was treated with corticosteroids. At follow-up after three months the EEG and the duplex scan were normal. There had been no TIAs since the start of the treatment. PMID- 8419842 TI - [Biological psychiatry]. PMID- 8419843 TI - [Prevention of recurrent erysipelas]. PMID- 8419844 TI - [Prevention of recurrent erysipelas]. PMID- 8419845 TI - [Prevention of recurrent erysipelas]. PMID- 8419846 TI - [The operation of an exercise-electrocardiographic service for family physicians; a description of 498 patients]. PMID- 8419847 TI - [Crib death and promethazine]. PMID- 8419848 TI - [Bacterial meningitis; pathogenesis and current possibilities of additional therapy]. PMID- 8419849 TI - [Decrease in the number of electroencephalograms in The Netherlands after 1975]. PMID- 8419850 TI - [Wrong questions, erroneous answers; a ridiculous choice]. PMID- 8419851 TI - Voice prosthesis in laryngectomee rehabilitation. AB - The use of a voice prosthesis is an acceptable option for the postlaryngectomy patient. Establishing criteria for patient selection is the most important way to avoid complications. The author describes a simple technique performed under local anesthesia. PMID- 8419852 TI - An American's perspective of Hungarian health care. AB - As U.S. health care affordability becomes a major issue, the author looks at the national health care system in Hungary. The statistics of Hungary's health care system are daunting: it is free, but not without cost to the patient. The system is being expanded and improved. PMID- 8419853 TI - Medical examinations under the Americans with Disabilities Act. AB - Title I of the Americans with Disabilities Act, effective July 26, 1992, established new guidelines for physicians who perform medical examinations of job applicants and existing employees with confidentiality requirements. These guidelines have a broad impact on physicians and employers. PMID- 8419854 TI - Early women physicians in New Jersey. AB - Dr. Madana DeHart was the first female graduate physician to practice in New Jersey. She was soon followed by Drs. Sarah DeHart, Florence DeHart, and Clara Krans. They had many barriers to overcome in their practices, but they succeeded. These women serve as role models to all physicians. PMID- 8419855 TI - Laparoscopic cholecystectomy in a community hospital. AB - The article reviews 400 laparoscopic cholecystectomies performed by the author in a community hospital, including patient profiles, workups, complications, and outcomes. The results indicate the benefits of laparoscopic cholecystectomy, including reduced morbidity and rapid recovery. PMID- 8419856 TI - Technologic cornucopias, the Bill of Rights, and slippery slopes. AB - Drug testing at the worksite is controversial. Drug testing offers the potential for massive erosion of Fourth Amendment rights. There are no easy answers to this dilemma, but the author provides some personal insight. Drug use testing should be balanced between societal needs and individual privacy rights. PMID- 8419858 TI - NPs and anesthesiologists: a collaborative practice model. PMID- 8419857 TI - Weight loss and OSA and pulmonary function in obesity. AB - Obstructive sleep apnea (OSA), daytime hypoxemia, and hypercapnia complicate obesity and are alleviated by weight loss. The flow-volume curve is a sensitive screening tool for most patients; the curve can monitor therapeutic efficacy of weight reduction. PMID- 8419859 TI - Using a stopwatch to assess pelvic muscle strength in the urine stream interruption test. AB - Pelvic muscle strength is important in maintaining urinary continence. The urine stream interruption test provides a simple measure of pelvic muscle strength. This study evaluated the accuracy of the test as adapted for clinical use. Women (n = 75) were tested according to standardized protocol. The test was simultaneously timed using a uroflowmeter (for research purposes) and a stopwatch (a technique more feasible in the practice setting). The stopwatch-timed urine stream interruption test was consistent with the uroflowmeter score (r = 0.90, p < 0.00) and demonstrated adequate repeatability (r = -0.69, p < 0.00). The stopwatch test was related to a digital measure of pelvic muscle strength (r = 0.49, p = 0.00), i.e., women with greater pelvic muscle strength were able to interrupt the stream of urine more quickly. Significantly less involuntary urine loss was seen in women whose stopwatch test score was two seconds or less as compared with those whose scores were greater than two seconds (t = -4.83, p = < 0.00, df = 73). Clinicians can use the urine stream interruption test as a baseline measure and as a tool to assess changes in pelvic muscle strength. PMID- 8419860 TI - 1992-93 update: how each state stands on legislative issues affecting advanced nursing practice. PMID- 8419861 TI - Federal reimbursement of advanced practice nurses' services empowers the profession. AB - In recent years, advanced practice nurses have achieved authority for the direct payment of their services under the four federal health programs--Medicare, Medicaid, Civilian Health and Medical Program of Uniformed Services, and the Federal Employee Health Benefit Plan. This article describes the importance of direct reimbursement and the barriers that have been overcome in achieving reimbursement. It also gives an overview of how advanced practice nurse services are covered and paid for in the federal programs. Increased authority has positioned nurses as visible members of the health care team. Direct payment for advanced practice nurses' services promotes public visibility for their role as direct providers of primary care and as managers of their patients' plan of care. Our nation is in an era of health care reform where additional primary care practitioners are needed. Recognition of advanced practice nurses allows them to be seen as a solution to our nation's pressing need for more primary care providers. PMID- 8419862 TI - Improper use of statistics is misleading. PMID- 8419863 TI - Vaginal yeast infections exacerbated by sugar intake. PMID- 8419864 TI - The nasopharynx in oral and maxillofacial radiology. II. Malignant lesions. AB - As with part I of this series of articles, this is not a comprehensive review of all malignant lesions to be found that involve the nasopharynx. It is an overview of how malignant lesions may appear and spread in the region of the nasopharynx. Oral and maxillofacial radiologists are reminded that familiarity with this area on diagnostic images ensures that lesions that involve this region will be interpreted as accurately as possible. PMID- 8419865 TI - Arthroscopic punch for definitive diagnosis of synovial chondromatosis of the temporomandibular joint. Case report and pathology review. AB - Synovial chondromatosis is characterized as a benign monoarticular condition with metaplastic cartilaginous nodules that develop within the synovial membrane of articulating joints. In addition to a pathology literature review, this article describes an unusual case of temporomandibular synovial chondromatosis that was sufficiently expansile to displace the condyle, which created not only a posterior occlusal apertognathia, but unusual articular bony fossa and eminentia erosions. An arthroscopic approach for definitive diagnostic punch biopsy and surgical approach is described and presented along with preoperative computerized tomograph, magnetic resonance imaging, and tomographic diagnostic images, as well as 3-year follow-up clinical and radiographic findings. Most interestingly, the arthroscopic examination and biopsy proved to be the most useful method to establish a definitive diagnosis of synovial membrane chondromatosis within the temporomandibular joint region for this patient. PMID- 8419866 TI - Digital subtraction temporomandibular joint tomography. AB - Receiver operating characteristics analysis was performed to demonstrate differences in diagnostic performance among conventional tomograms, digitized tomograms, and subtraction tomograms. Digital subtraction tomography was found to be the best imaging modality for detecting artificially created lesions in the two selected temporomandibular joint locations. There was a statistically significant difference in diagnostic performance between conventional tomograms and subtraction tomograms for the detection of temporomandibular joint bony lesions. There was also a statistically significant difference in diagnostic performance between digitized tomograms and subtraction tomograms in the detection of these bony lesions. There was no statistically significant difference in diagnostic performance between conventional tomograms and digitized tomograms for the detection of temporomandibular joint bony lesions. The significance level was set at p = 0.05. Results of the analyses of variance showed that with digital subtraction tomograms, observer reliabilities were higher than with conventional and digitized tomograms. PMID- 8419867 TI - The pig as an animal model for experimentation on the temporomandibular articular complex. AB - A macroscopic study of the so-called temporomandibular joint of omnivorous mammals (pigs), carnivores (dogs and cats), rodents (rabbits and rats), and herbivores (cows, sheep, and goats) revealed the existence of two left and right temporomandibular articular complexes, each made up of a condylomeniscal and a meniscotemporal joint on either side, as in human beings. In all cases there is a combination of a condylar joint and a reciprocally fitting one. We conclude that the pig is the experimental animal model that most closely resembles human beings for the study of the cited articular structures. PMID- 8419868 TI - Postsurgical temporomandibular joint hypomobility. Rehabilitation technique. AB - Specific techniques to treat temboromandibular joint (TMJ) hypomobility caused by capsular restriction are explained. Initially inflammation must be controlled. TMJ manipulation by condylar distraction during opening, protrusion, and lateral movements, and a simple stretching exercise to maintain increased mandibular range of motion, are described. Resistive opening and closing exercises at full opening to relax the lateral pterygoid muscles are prescribed. For all exercises five repetitions, repeated five times per day, are prescribed. These techniques are demonstrated in the successful treatment of a child with a presurgically and postsurgically hypomobile right TMJ. PMID- 8419869 TI - Tranexamic acid as a mouthwash in anticoagulant-treated patients undergoing oral surgery. An alternative method to discontinuing anticoagulant therapy. AB - A double-blind randomized study was carried out to evaluate the clinical hemostatic effect of tranexamic acid mouthwash after dental extraction in 30 patients who received anticoagulant agents. Surgery was performed with a reduction in the level of anticoagulant therapy in the control group and with no change in the level of anticoagulant therapy in the group who received the tranexamic acid. After the extraction the surgical field was irrigated with a 5% solution of tranexamic acid in the group of 15 patients whose anticoagulant treatment had not been discontinued and with a placebo solution in the group of 15 patients for whom the anticoagulant therapy was reduced. Patients were instructed to rinse their mouths with 10 ml of the assigned solution for 2 minutes four times a day for 7 days. There was no significant difference between the two treatment groups in the bleeding incidence after oral surgery. We conclude that the anticoagulant treatment does not need to be withdrawn before oral surgery provided that local antifibrinolytic therapy is instituted. PMID- 8419870 TI - Successful pain management by Finnish oral surgeons. PMID- 8419871 TI - Absorbed dose determination for tomographic implant site assessment techniques. PMID- 8419872 TI - Presenting signs and symptoms of nasopharyngeal carcinoma. AB - Pain in the head and neck may be due to multiple causes and difficult to localize. A definitive diagnosis can be difficult to make. The literature reports several cases of nasopharyngeal carcinoma presenting as temporomandibular disorders (TMD). The records of 52 patients with a diagnosis of nasopharyngeal carcinoma were reviewed to determine the frequency of symptoms that may be similar to TMD. In 13.5% of the patients common TMD signs and symptoms were present, such as clicking in the joint, pain while chewing, and limited opening. Of the patients reviewed, 44.2% described the location of their pain as headache, earache, or as jaw, midface, or neck pain. The most common description of pain was aching, dull, or pressing. Symptoms identified that may assist in differentiation of nasopharyngeal carcinoma from TMD include nosebleed, nasal stuffiness, altered hearing, and cervical lymph node enlargement. A patient with carcinoma of the nasopharynx may have symptoms that must be differentiated from TMD. PMID- 8419873 TI - Oral complications in children with cancer. AB - Oral complications during cancer therapy are a common source of discomfort and a potential source of systemic infection. We report the results of a 2 1/2-year prospective follow-up study on the incidence of oral complications in 214 pediatric patients with cancer. Overall, the incidence of ulcers in these patients ranked highest followed by gingivitis. Children with sarcomas had more ulcers (p = 0.03) and Candida infections (p = 0.03) than those with leukemia. The rate of gingivitis among patients with leukemia was five times higher than in patients with sarcoma (p = 0.02). Candida infections in children with solid tumors occurred four times more often than in patients with leukemia (p = 0.02). This study shows that oral complications are a frequent cause of morbidity in children with cancers and are more common in some cancers than in others. Oral complications may be prevented or diminished in severity by identifying the risk groups and developing preventive and treatment strategies. PMID- 8419874 TI - Hereditary hemorrhagic telangiectasia with florid osseous dysplasia. Report of a case with differential diagnostic considerations. AB - The clinical presentation of hereditary hemorrhagic telangiectasia with various manifestations has been well described as has florid osseous dysplasia. There have been no cases reported of the two pathologic entities in the same patient. We present a case with the simultaneous occurrence of hereditary hemorrhagic telangiectasia and florid osseous dysplasia with important considerations for differential diagnosis, and we discuss whether this case presents a potential syndrome. PMID- 8419875 TI - Decortication in the treatment of diffuse sclerosing osteomyelitis of the mandible. Retrospective analysis of 41 cases between 1969 and 1990. AB - Thirty-four patients with mandibular diffuse sclerosing osteomyelitis who had been treated by means of 61 decortications were evaluated retrospectively. Eighteen patients (53%) were free from symptoms on an average of 5.4 years after surgery. Of these, 12 had improved after their first operation. In the other six patients, decortication was performed two to four times before healing was clinically observable. Symptoms recurred in 75% of the cases within 12 months after surgery. Neither sex, location, extent, and chronicity of the disease nor the precise surgical technique used seemed to affect the outcome. The patients who exhibited improvement, however, were significantly older and more often edentulous than the patients in whom the symptoms recurred. Possible causes of failure were an insufficiently radical surgical procedure and retention of devitalized teeth in the decorticated area. PMID- 8419876 TI - Epidermolysis bullosa. A case report. AB - Epidermolysis bullosa is a group of rare genetic-related skin disorders. It is characterized by bullae and vesicles on the skin and mucosa, that result from friction, trauma, or heat. This article reports a case of Epidermolysis bullosa. With proper diagnosis, the dentist can treat a patient with this type of disorder without causing bullae as a result of treatment. PMID- 8419877 TI - Search for correlation of radon levels and incidence of salivary gland tumors. AB - Studies were undertaken to determine if there is a statistical correlation between radon levels and the incidence of salivary gland tumors because of high levels of radon in Pennsylvania. In Part I of the study, the incidence of minor salivary gland tumors accessioned by Temple University, Emory University, and University of Southern California from 1986 to 1988 were correlated with average radon levels in the three locations with the use of standard statistical analyses. In Part II, the occurrence of malignant salivary gland tumors was obtained for each of the 67 counties in Pennsylvania from 1986 to 1988 and correlated statistically with radon levels and population figures in each of those counties. A statistically significant correlation between radon levels and incidence of salivary gland tumors could not be demonstrated in either case. PMID- 8419878 TI - Vascular neoplasms of the parotid gland. Parotid vascular tumors. AB - Vascular neoplasms of the parotid gland are common in early childhood, particularly in females. We reviewed the clinical, histologic, and treatment details of 10 cases of hemangiomas and one case of lymphangioma that involved the parotid gland. Histologically, the cellularity and increased division figures in these lesions should not be interpreted as a sign of a malignant condition. A watchful expectancy for spontaneous regression and preservation of the facial nerve at surgery are advocated. PMID- 8419879 TI - Trichodysplasia and amelogenesis imperfecta. AB - This paper describes a family in which members of two generations have an X linked type of enamel dysplasia. All affected persons have symmetric pits in the cuticles of their hair shafts. The observation of these concurrent traits raises questions about the classification of amelogenesis imperfecta and the value of microscopic studies of the hair in the persons with purported isolated enamel dysplasias. PMID- 8419880 TI - Effect of ArF-193 nm excimer laser on human dentinal tubules. A scanning electron microscopic study. AB - The purpose of this study was to evaluate the effects of the ArF-193 nm excimer laser on the dentinal tubules of extracted human teeth under a scanning electron microscope. Fifteen 3 mm thick slices were cut with an electric saw at the cementoenamel junction from 15 extracted human teeth. A diamond bur was used to remove the cementum layer and expose the dentinal tubules. Each slice was scored by a permanent marker into four equal quadrants. The ArF excimer laser was applied for 5 seconds on three of the quadrants with fluences that ranged from 0.2 J/cm2 to 15 J/cm2 and pulse repetition of 25 Hz. The untouched quadrant served as a control. The specimens were mounted on stubs, sputter coated by gold, and examined by a scanning electron microscope. The effects of the ArF excimer laser irradiation varied. Laser fluences of 0.2, 0.5, and 1.0 J/cm2 had no effect. Although fluence of 15 J/cm2 caused significant removal of peritubular dentin, melting and resolidification of the dentinal smear layer was also observed under the scanning electron microscope with a laser fluence of 5 J/cm2. PMID- 8419881 TI - Is phantom tooth pain a deafferentation (neuropathic) syndrome? Part I: Evidence derived from pathophysiology and treatment. AB - Phantom tooth pain is a syndrome of persistent pain or paresthesia in teeth and other oral tissues that may follow dental or surgical procedures such as pulp extirpation, apicoectomy, tooth extractions, or exenteration of the contents of the maxillary antrum. It can also occur when nerves are injured after trauma to the face or even after routine inferior alveolar nerve blocks if the needle pierces the nerve sheath. In the case of tooth extraction, the pain is found in the edentate area. After periodontal surgery, pain or paresthesia is located in the gingiva. The incidence of phantom tooth pain after extirpation may be as high as 3% of cases. Clinically, phantom tooth pain is similar in many essential characteristics to deafferentation pain syndromes also known as phantom pain syndromes. A limitation to this taxonomy is the lack of definitive information with respect to the pathophysiology of deafferentation pain in the trigeminal nerve. This article amplifies previous clinical descriptions of phantom tooth pain. Current concepts in the pathophysiology of neuropathic pain are reviewed as they pertain to phantom tooth pain. Treatments are described that use three routes of drug administration: oral, nerve blocks by injections, and intranasal applications. Reasons are discussed for the high rates of morbidity after dental and neurosurgery in attempts to treat phantom tooth pain. PMID- 8419882 TI - [Immunotherapy of human tumors]. PMID- 8419883 TI - [Fibrogenesis in the liver--fibrosis, cirrhosis]. AB - Liver fibrogenesis is a delicately balanced process, in which mainly the non parenchymal liver cells are implicated. Either increased synthesis or decreased catabolism of matrix proteins results in the enhancement of ECM. As further consequence the formation of continuous diffusion and filtration barriers along the Disse space will hinder the bidirectional exchange of macromolecules. Normal structure of ECM is necessary to the normal function of hepatocytes. The quantitative and qualitative changes of ECM observed in liver fibrosis are able to inhibit the liver specific functions of hepatocytes. The mechanisms involved in this effect are not yet clearly understood. In animal experiments liver cirrhosis is reversible and theoretically the chance is open for humans, as well if we will be able to influence the specific steps of fibrogenesis. PMID- 8419884 TI - [Connection between painless abnormal ST-deflection and autonomic neuropathy in diabetes mellitus]. AB - In order to assess the relationship between abnormal but silent ST-segment depression and autonomic neuropathy 63 diabetic patients (age: 40-71 years, duration of diabetes: 2-32 years) without a history of angina pectoris were investigated. Transient ST-segment depression was assessed by 24 hours Holter monitoring and, in addition, dynamic exercise on bicycle ergometer was also performed in all but 7 patients. Autonomic neuropathy was evaluated by cardiovascular function tests (deep breathing, Valsalva manoeuvre and lying-to standing). Abnormal ( > or = 2 mm) ST segment depression was observed in 11 patients (18%) while signs of autonomic neuropathy were found in 37 diabetics (59%). Signs of autonomic neuropathy were significantly (p < 0.01) more often documented in patients with (11/11) than without (26/52) abnormal ST-segment depression. It was concluded that autonomic neuropathy could be a possible explanation for lacking symptoms from abnormal ST-segment depression in diabetic patients. PMID- 8419885 TI - [Importance of evaluating clinical symptoms of alcoholic intoxication]. AB - The authors studied the relationship between the blood alcohol concentration and the categories of alcohol intoxication established by means of examining the clinical signs of intoxication in a drinking test, and by analyzing the reports on driving while intoxicated (DWI). In the drinking test the relative increase of viso-motor reaction time was an accurate indicator of the blood alcohol levels. In almost half of the cases there was no correlation between the clinical signs and the blood alcohol concentration. When alcohol and medicine was used in combination a more severe intoxication was frequently assumed. Therefore, if the reported intoxication based on clinical signs indicates a more severe intoxication than that correlating with the blood alcohol concentration, the toxicologic analysis of the blood samples are unconditionally necessary. PMID- 8419886 TI - [An unusual case of intercalary type of limb abnormalities]. AB - Authors report about a case of total lack of middle phalanxes on the 3d and 4th fingers and a hypoplasia of the surrounding phalanges on the left hand. This is the first report on this type of intercalary type in congenital limb deficiency group. They call attention to one of the so far not sufficiently emphasized hazard of previous periconceptional oral contraceptive use. If there is not enough time left for the total regeneration after the discontinuation of contraceptives, such kind of malformation may develop in the fetus due to the insufficiency of fetal-placental circulation. PMID- 8419887 TI - [Recommendation on the definition of the nosocomial character of infectious manifestations]. AB - The definitions describing when and which infectious diseases can be considered of nosocomial origin have been missing. The lack of such definitions make impossible the exact evaluation of their frequency and position taking for their prevention. This recommendation contains definitions based on the epidemiological characteristic of more than 50 nosocomial infectious diseases. Such an effort is urgently needed because without uniform definitions a successful surveillance of nosocomial infections remains unrealistic. The proposed definitions are subject for public and personal discussion. PMID- 8419888 TI - [In memory of a forgotten cardiologist. Laszlo Unghvary 1904-1972]. PMID- 8419889 TI - [Universities and medical education in Central and Eastern Europe]. PMID- 8419890 TI - You don't have to be born a leader to become one. PMID- 8419891 TI - Safeguarding the rights of people who have had cancer. PMID- 8419892 TI - Obstructive apnea, associated patterns of movement, heart rate, and oxygenation in infants at low and increased risk for SIDS. AB - Repetitive polysomnograms were recorded between 40 weeks post-conceptional age and 6 months in a total of 49 infants, 19 healthy preterm infants, 14 normal term infants, and 16 subsequent siblings of infants who died of sudden infant death syndrome (SIDS). These nighttime recordings lasted 2-4 hours, except at 3 months when an overnight 12-hour recording was performed. Obstructive apneas (OA) > 3 seconds were divided into 3 categories: 1) clear obstructive, 2) mixed and 3) unclear because of movement artifacts. More than half belonged in category 3 and were excluded from further analysis unless accompanied by a transient episode of bradycardia (TEB), defined as heart rate < or = 100 beats per minute. Each OA with TEB was also examined for changes in transcutaneous oxygen tension (PtcO2). Most pauses were brief (median, 4 seconds), the longest (27 seconds) seen only once in the youngest premature infant. The majority of OA were accompanied by heart rate accelerations. The number of clear obstructive and mixed apneas was similar. The scores were combined to calculate a density (number per 100 minutes of recording). OA were not common: Their density decreased from 2 in 100 minutes at 40 weeks in the preterm to once every 300 minutes (5 hours) in the 6-month-old term infant. Ten percent of the OA were accompanied by TEB. Of these, 10% were accompanied by a PtcO2 decrease of > 10 mm Hg. OA with TEB followed a nonmonotonic curve, the highest percentage of infants showing this pattern at the age of highest risk for SIDS. Minor differences among study groups were confined to less movements with OA in subsequent siblings and an earlier peak incidence of OA with TEB in prematures, compared to normal term infants. OA were seen in all study groups, were self-limited, and apparently were devoid of pathological consequences. PMID- 8419893 TI - Lung recoil and the determination of airflow limitation in cystic fibrosis and asthma. AB - A reduction of lung recoil pressure could aggravate any airflow limitation that might be present in patients with cystic fibrosis (CF) or asthma. In a group of 22 children and young adults (8 with cystic fibrosis, 8 with asymptomatic asthma, and 6 healthy controls) aged 8-24 years, lung recoil pressure (Pst) at 100%, 90%, and 60% of TLC and static lung compliance (Cst) were measured using an esophageal balloon. The indices of airflow limitation, including maximal expiratory flow at 25% VC (Vmax25), forced expiratory volume in 1 second (FEV1), and specific airway conductance (sGaw), were also measured. In all patients, Vmax25 was reduced, the airway obstruction being more pronounced in patients with CF. Pst was reduced in CF and asthma, again more in the patients with CF. Cst was normal in both groups because the pressure volume curve was shifted up and to the left. There were significant correlations between Pst at 60%, 90%, and 100% of TLC and both Vmax25 and FEV1 (P < 0.01). sGaw correlated with Pst90 and Pst60 (r = 0.47 and 0.53, respectively; P < 0.05 for both). No correlation was found between Cst and Pst at any lung volume. No correlations were observed between Cst and Vmax25, FEV1, or sGaw. These results suggest that loss of elastic lung recoil pressure is a factor in airflow limitation of children and young adults with CF or asthma. PMID- 8419894 TI - The relationship of RSV-specific immunoglobulin E antibody responses in infancy, recurrent wheezing, and pulmonary function at age 7-8 years. AB - In order to determine if respiratory syncytial virus (RSV)-specific IgE responses at the time of bronchiolitis in infancy are related to recurrent wheezing and pulmonary function at 7-8 years of age, a cohort of 43 infants was identified at the time of their initial RSV bronchiolitis episode. RSV-specific IgE responses in nasopharyngeal secretions were determined, and patients were then followed prospectively with notation of the number of wheezing episodes and exposure to cigarette smoke at home. At 7-8 years of age the patients underwent skin testing to 7 environmental allergens and pulmonary function testing, including pulse oximetry and methacholine challenge. Pulmonary function following inhalation of bronchodilating agents was compared to baseline pulmonary function results in order to determine if abnormalities of pulmonary function were reversible. Recurrent wheezing following bronchiolitis was associated with the initial RSV IgE response, as well as with a family history of asthma. Current wheezing at age 7-8 years was associated with 2 or more positive skin tests (P < 0.01), a history of exercise-induced wheezing (P = 0.01), and increased sensitivity to methacholine (P < 0.01). Pulmonary function test results were similar for groups with and without recurrent wheezing following bronchiolitis. For the entire study group, RSV-IgE specific responses were unrelated to pulmonary function, but small airway dysfunction was associated with passive smoking (P < 0.025), and both large airway dysfunction and increased airway reactivity were associated with the number of positive skin tests (P < 0.025). Reduced small airway function improved following bronchodilator inhalation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419895 TI - Acoustic vs. spirometric assessment of bronchial responsiveness to methacholine in children. AB - To study wheezing as an indicator of bronchial responsiveness during methacholine challenge (MC) in children, we used computer analysis of respiratory sounds and compared wheeze measurements to routine spirometry. MC was performed in 30 symptomatic subjects (sympt), age 11 +/- 3.1 years (mean +/- SD), with suspected asthma and in 12 controls (contr), age 10 +/- 3.4 years. Respiratory rate (RR), spirometry, arterial oxygen saturation (SaO2), and cough were registered until the concentration provoking a > or = 20% fall in forced expiratory flow in 1 second (FEV1;PC20), or the end point (8 mg/mL) was reached. For 1 min after each inhalation, sounds over the trachea and posterior right lower lobe were recorded together with calibrated airflow. Computer analysis of respiratory sounds was used for objective wheeze quantification. Wheezing was measured as its duration relative to inspiration (Tw/Ti) and expiration (Tw/Te). Seventeen of the sympt group developed wheezing (sympt/W) with > or = 5% Tw/Ti or > or = 5% Tw/Te. Thirteen of the sympt did not wheeze (sympt/no W). Three contr developed wheeze (contr/W) while 9 did not (contr/no W). In sympt/W, RR increased from 20 +/- 6.2 per min at baseline to 25 +/- 9.2 (P < 0.05) at the MC concentration provoking wheeze (PCw), and SaO2 decreased from 97.4 +/- 1.2% to 95.3 +/- 2.4 (P < 0.05). In contr/W, RR did not change, but SaO2 decreased from 97.3 +/- 1.5% to 95.7% +/- 1.2% (P < 0.05). Wheezing occurred at both recording sites and was as common during inspiration as during expiration.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419896 TI - Pulmonary mechanics and gas exchange: effect of lateral positioning during recovery from respiratory distress syndrome. AB - Sixteen stable intubated premature infants without a clinically significant patent ductus arteriosus were studied during recovery from respiratory distress syndrome in order to determine the effects of left and right lateral, as compared to supine, positioning. Pulmonary mechanics were measured for spontaneous breaths 5 and 15 minutes after positioning, and arterial blood gases 15 minutes after positioning. Infants were randomized to 1 of 2 position sequences: (1) supine, left, supine, right or (2) supine, right, supine, left. No significant differences were detected between positions for dynamic compliance, tidal volume/kg, and total, inspiratory and expiratory pulmonary resistance. Likewise, no significant differences in PaO2 or PaCO2 were detected between the positions. The sequence of positions did not affect the pulmonary mechanics of spontaneous breaths or arterial blood gases. This suggest that short-term lateral positioning as well as supine positioning can be utilized without deleterious effects on pulmonary mechanics and gas exchange in neonates recovering from respiratory distress syndrome. PMID- 8419897 TI - Assessment of exercise capacity in asthmatic children with various degrees of activity. AB - Physical fitness in a group of 49 stable asthmatic children was determined by an incremental exercise test. Thirty-one normal children served as a control group. The asthmatic children were divided into three groups. Group 1 was comprised of 16 children who actively participated in organized sports, Group 2 of 16 children who did not participate in organized sports but who engaged in free-play, and Group 3 of 17 children with a sedentary life-style who avoided even free-play. The results of cardiopulmonary evaluation before and after maximal incremental exercise testing have shown that Groups 1 and 2 behaved like the control group and their physical fitness was similar. Group 3 whose life-style was sedentary had poor physical fitness as compared to the other asthmatics and to the control group. This was the result of poor cardiovascular conditioning and was unrelated to the respiratory limitation. We conclude that poor physical fitness in asthmatic children is the result of a sedentary life-style and can be potentially normalized. PMID- 8419898 TI - Aerodynamic and laryngographic assessment of pediatric vocal function. PMID- 8419899 TI - Complications following oat head aspiration. AB - We report 5 cases of oat head aspiration in children that resulted in serious complications due to the unidirectional migration of the oat head to the periphery of the lung. The complications included pneumothorax, pneumomediastinum, recurrent hemoptysis, chronic lung disease, bronchiectasis, lobectomy, bronchopleural and bronchocutaneous fistulae, pleural effusion, empyema cavity, and, one not described before, osteomylitis of the rib. Physicians should be aware of the dangers with this particular foreign body aspiration. PMID- 8419900 TI - Papillary cystic tumor of the pancreas: a case indistinguishable from oncocytic carcinoma. AB - A case of papillary cystic tumor (PCT) of the pancreas in a 55-year-old woman is described. She presented with a gradually enlarging and painless abdominal mass for > 26 years. The tumor was encapsulated and measured 16 x 12 x 10 cm. The gross features and conventional light microscopic appearance of this tumor were consistent with the previously reported cases of PCT. Perineural and capsular invasions were found. In addition to the densely packed blue granules in the cytoplasm demonstrated by phosphotungstic acid-hematoxylin stain, ultrastructural study revealed the tumor cells to be packed with numerous mitochondria. These oncocytes comprised almost all the non-necrotic tumor areas. Therefore, this case was indistinguishable pathologically from oncocytic carcinoma of the pancreas. DNA flow cytometry showed a diploid pattern and low S phase fraction, indicating that the tumor has a low proliferative activity and a favorable prognosis. PMID- 8419901 TI - Sequential invasions of pancreatic pseudocysts in pancreatic tail, hepatic left lobe, caudate lobe, and spleen. AB - A 66-year-old male patient without a history of risk factors for pancreatitis suffered from pancreatitis and developed pseudocyst. During the course of treatment and follow-up, the pseudocyst was found to have migrated through the pancreatic tail, left hepatic lobe, caudate lobe, and spleen on abdominal sonography and computed tomography scan. Finally, emergent laparotomy was done for splenic abscess and removal of infected pseudocyst in the spleen and lesser sac of the abdomen. The patient made a full recovery after operation. PMID- 8419902 TI - Pancreatic tubercular abscess. PMID- 8419903 TI - Dynamic in vivo observation of rat islet microcirculation. AB - In vivo fluorescent microscopy with direct observation of flow through the islet was used to investigate the islet microcirculation. In urethane-anesthetized rats (n = 18), the pancreas was exposed and an islet was identified under direct microscopy. The vertical illuminator for fluorescent microscopy was turned on and fluorescein-albumin conjugate or fluorescent microspheres were injected intravenously or intraarterially. Each study was videotaped; on slow motion playback, the flow of the conjugate or microspheres was followed through the islet, the islet capillaries, and then to venules exiting the islet. One islet in the head of the pancreas in 12 rats was studied. The arterioles first reached the surrounding mantle of the islet where they divided into capillaries that carried conjugate or microspheres to other portions of the mantle or the core of the islet. Flow of conjugate traversed the core and returned to different portions of the mantle. The fluorescent microsphere study permitted a more detailed study of the pathways followed, the individual microspheres being seen to travel through numerous tortuous pathways through the islet. The flow of microspheres was nonhomogeneous in that individual microspheres in one portion of the islet would stop, then move on, while other microspheres flowed freely. The capillaries joined two to six venules that carried the conjugate or microspheres out of the islet. One or two of the exiting microvessels entered the adjacent acinar microcirculation; the others entered larger collecting venules. In six tail islets studied, the microcirculation was similar to that of the islets in the head of the pancreas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419904 TI - Is the hydrogen clearance technique a useful tool for measurements of pancreatic blood flow during acute experimental pancreatitis? AB - Changes of pancreatic blood flow (PBF) during acute pancreatitis (AP) have been under investigation by means of electromagnetic flowmeters, radioactive microspheres, isotope fractionation, radioactive gas clearance, and venous outflow techniques. All methods, however, have certain drawbacks, which make the application of other techniques desirable. In this study, the hydrogen clearance technique (HCT) was tested for the first time in a well-established foxhound model of AP. PBF, systemic blood pressure, and heart rate were monitored over 90 min after the onset of AP and 60 min after therapeutic infusion of Dextran-40 (10 ml/kg body wt). Our results fully agree with the data found by other techniques in this experimental model. Sixty-five of 73 electrodes implanted into the pancreas of eight foxhounds were found working. From 1,024 registered clearance curves, 876 were identified as monoexponential. In the other cases, and only then, we found either dislocation of the electrode tips (n = 6) or perielectrodal hemorrhage during histological examination (n = 4). We believe that the HCT is a feasible and reliable tool for measuring PBF in experimental settings like AP. PMID- 8419905 TI - Microcirculatory response of the pancreas to feeding, sham feeding, and truncal vagotomy in conscious dogs. AB - In this study, changes in the microcirculatory dynamics of the pancreas in response to normal feeding, sham feeding, and truncal vagotomy were investigated to elucidate the involvement of a neural mechanism in the physiological modulation of pancreatic blood flow in the conscious state. Continuous measurement of changes in the microcirculation of the pancreas was performed in conscious dogs by the thermoelectric method. A meat meal was given to six normal dogs, seven dogs constructed with external esophagostomy (sham feeding), and four dogs with truncal vagotomy. In response to normal feeding, pancreatic blood flow attained the peak increase of 65.2 +/- 6.2%, showing a significant and biphasic response until approximately 120 min. After sham feeding, pancreatic blood flow was significantly increased with peak values of 89.0 +/- 19.0%, but thereafter showed a rapid decrease, returning to the basal level already at 7.2 +/- 1.1 min. Although truncal vagotomy significantly and greatly reduced the peak increase of pancreatic blood flow to 28.2 +/- 5.1%, blood flow showed still a significant and sustained elevation above basal. This study provides evidence for the involvement of the neural mechanism in the physiological modulation of the microcirculation of the pancreas in the conscious state. The results strongly suggest that the cephalic phase of the increase in pancreatic blood flow is vagally mediated. PMID- 8419906 TI - Release of catecholamines is increased but does not contribute to the impaired insulin secretion in the perfused pancreata of diabetic rats. AB - Insulin response to glucose is severely impaired in patients with non-insulin dependent diabetes mellitus (NIDDM). Also in a rat model of NIDDM, neonatally streptozotocin diabetic rats (STZ), the insulin response to glucose is profoundly suppressed when studied in vivo or in the perfused pancreas. The insulin response was better preserved from isolated islets obtained from patients and STZ rats. Since alpha 2-adrenoceptor stimulation suppresses insulin secretion, catecholamines from intrapancreatic nerve terminals may be involved in the mechanism behind the marked impairment of the glucose-stimulated insulin response in the intact pancreas. We have studied the pancreatic content and release--or overflow--of catecholamines from the isolated, perfused pancreas of STZ rats. The overflow of noradrenaline (NA) in the perfusate was two- to threefold higher in pancreata from STZ than from nondiabetic rats, and perfusion with a high glucose concentration increased the NA overflow in both types of pancreata. Levels of adrenaline (ADR) were always low in perfusates of nondiabetic glands, but increased in five of seven perfusions of STZ glands. The pancreatic contents of NA and ADR were similar in STZ and nondiabetic rats. Pretreatment of rats with reserpine 24 h before perfusions reduced the pancreatic content of NA and ADR by > 90% in both STZ and nondiabetic rats. Reserpine also diminished the overflow of NA in the perfusate from STZ rats by > 90% and from nondiabetic rats by 58-77%. After reserpine, however, glucose-induced insulin release was not enhanced in either STZ or control pancreata. In conclusion, overflow of catecholamines is higher in the pancreas of STZ than of nondiabetic rats.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419907 TI - Dual effects of gastric inhibitory polypeptide on insulin secretion. AB - The role of gastric inhibitory polypeptide (GIP) on insulin secretion in the presence of different glucose concentrations has been studied in perifused microdissected murine islets. Insulin secretion was concentration dependent in the presence of glucose alone: Switching the perifusion buffer from 5.5 to 11.1 and 22.2 mM glucose caused an increase in insulin response assessed as the total integrated area under the curve over a 20-min period (6.4 +/- 0.48 and 12.1 +/- 0.58 ng, respectively; p < 0.01, n = 6). If 11.1 mM glucose perifusion in the presence of GIP was preceded by 5.5 mM glucose alone, the integrated insulin secretion/20 min above basal level was attenuated (1.46 +/- 0.10 vs. 0.37 +/- 0.03 ng; p < 0.01, n = 6), and withdrawal of GIP from the perifusion buffer resulted in the restoration of 11.1 mM glucose-stimulated insulin secretion (1.46 +/- 0.10 vs. 1.98 +/- 0.12 ng). If islets were continuously perifused with 11.1 mM glucose, the addition of GIP did not alter insulin secretion. In contrast, the addition of GIP to 22.2 mM glucose perifusion buffer further enhanced the high glucose-induced insulin secretion above basal (12.1 +/- 0.58 vs. 14.5 +/- 0.84 ng; p < 0.05, n = 6). These observations are consistent with a hypothesis that during a low glucose condition, GIP prevents the risk of hypoglycemia by suppressing insulin secretion, while during a high glucose load, glucose-induced insulin stimulation is potentiated by GIP, presumably to prevent hyperglycemia. PMID- 8419908 TI - Differences in K+ permeability between cultured adult and neonatal rat islets of Langerhans in response to glucose, tolbutamide, diazoxide, and theophylline. AB - The effects of glucose, tolbutamide, and diazoxide on K+ permeability in neonatal and adult rat pancreatic islets, maintained in culture 1 week, were investigated by measuring the 86Rb outflow rate from prelabeled islets. In the absence of glucose, the 86Rb efflux was significantly lower in neonatal than adult islets. Raising the glucose concentration to 2.8, 5.6, 8.3, and 11.1 mM produced a marked reduction in the 86Rb efflux in adult islets but only a minor reduction in neonatal islets. The effect of tolbutamide to reduce, and diazoxide to increase, the 86Rb efflux was also less in neonatal islets. These results are discussed with respect to previously reported differences in insulin secretion from neonatal and adult islets in culture. PMID- 8419909 TI - Long-term diabetogenic effect of streptozotocin in rats. AB - The long-term effects of streptozotocin (30-70 mg/kg) were studied on plasma glucose and insulin levels, islet morphology, and glucose-stimulated insulin secretion in rats. In addition, the protective effect of short-term (7 days) insulin treatment on streptozotocin-induced diabetes was examined. Streptozotocin administration at dose levels exceeding 40 mg/kg resulted in a long-term, stable hyperglycemia with no insulin response to glucose at 3 months and with a marked derangement of islet morphology (few insulin cells, accumulation of glucagon cells). In contrast, at 30 and 40 mg/kg, streptozotocin induced a transient diabetes. Thus, the blood glucose levels, being elevated at days 1-7, returned to normal levels within 10 days after streptozotocin administration and the glucose induced insulin secretion, being absent at day 1, was normal at 3 months. Furthermore, the islet morphology was also normal in these groups at 3 months. Short-term (7 days) insulin treatment normalized the long-term diabetes in rats given 50 mg/kg streptozotocin, but not in rats given 60 or 70 mg/kg streptozotocin. Thus, after insulin treatment, all rats receiving 50 mg/kg streptozotocin returned to normoglycemia within the following 2 weeks, and the glucose-induced insulin secretion was normal after 3 months, as was islet morphology.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419910 TI - Cortivazol increases glucocorticoid receptor expression and inhibits growth of hamster pancreatic cancer (H2T) in vivo. AB - Glucocorticoids are effective in the treatment of certain leukemias and lymphomas, but their effects on the growth of several solid tumors have not been determined. We report here that cortivazol (CVZ), a potent synthetic glucocorticoid, inhibits the growth of a hamster pancreatic adenocarcinoma, H2T, in vivo. CVZ regulation of glucocorticoid receptor (GR) expression was followed as a specific molecular correlate. H2T cells were injected into cheek pouches of male Syrian golden hamsters, where they formed readily measurable tumors. Two studies were performed. In the first, hamsters were randomized to three groups immediately after injection of tumor cells: control, CVZ (0.1 micrograms/g body wt), or CVZ (0.3 micrograms/g body wt). Injections of either CVZ or its vehicle were administered on a 14-day cycle of 5 treatment days, followed by 9 days off treatment. Tumors were measured and areas calculated weekly. On day 48, the hamsters were killed and the tumors excised, weighed, and analyzed for DNA, RNA, and protein content. In the second study, randomization and treatment schedule were as before, except that on day 33 the hamsters were killed, tumors were excised and weighed, and total RNA from the tumors was isolated. GR mRNA content was determined by filter hybridization with a 32P-labeled GR cDNA probe, and the signal normalized by reprobing for alpha-tubulin as an invariant, independent signal. At either dose, CVZ significantly inhibited H2T tumor area and weight and DNA, RNA, and protein content. Body weights of animals treated with CVZ were not significantly decreased as compared with controls. In addition, GR mRNA in H2T cells was increased approximately twofold by CVZ.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419911 TI - Role of phospholipase A2 in pancreatic acinar cell damage and possibilities of inhibition: studies with isolated rat pancreatic acini. AB - Phospholipase A2 (PLA2) has been postulated to play an important role in the pathogenesis of acute pancreatitis. To study the mechanism through which PLA2 may cause cellular damage, we used an in vitro model of isolated rat pancreatic acini prepared by collagenase digestion. Newly synthesized proteins were labeled by [35S]methionine. Cellular destruction was measured by the degree of release of radiolabeled proteins. Incubation of pancreatic acini with PLA2 alone caused only minor damage when very high concentrations of this enzyme were used. However, when acini were incubated with PLA2 in combination with its substrate, lecithin, cells were destroyed in a time- and concentration-dependent manner. Incubating cells with pancreatic homogenates and lecithin caused damage only when there had been prior activation of homogenates with either trypsin or enterokinase. The damage could be simulated by incubating acini with pure lysolecithin. Alcohol and cerulein did not further increase the destruction caused by PLA2 and lecithin. When acini were incubated with supernatants from another set of acini to which oleic acid had been added, a similar degree of damage resulted as compared with acini incubated with oleic acid alone. However, adding PLA2 to supernatants from acini preincubated with fatty acids significantly increased the degree of cellular necrosis. The destruction by PLA2 and lecithin was inhibited by albumin but could not be inhibited by gabexate mesilate, nafamostat mesilate, or cytidine diphosphocholine. We conclude that PLA2 could play a role in pancreatic acinar cell damage, especially in the spread of cellular necrosis within the organ, provided that its substrate, lecithin, is present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8419912 TI - Perturbation of the secondary structure of the scrapie prion protein under conditions that alter infectivity. AB - Limited proteolysis of the scrapie prion protein (PrPSc) generates PrP 27-30, which polymerizes into amyloid. By attenuated total reflection-Fourier transform infrared spectroscopy, PrP 27-30 polymers contained 54% beta-sheet, 25% alpha helix, 10% turns, and 11% random coil; dispersion into detergent-lipid-protein complexes preserved infectivity and secondary structure. Almost 60% of the beta sheet was low-frequency infrared-absorbing, reflecting intermolecular aggregation. Decreased low-frequency beta-sheet and increased turn content were found after SDS/PAGE, which disassembled the amyloid polymers, denatured PrP 27 30, and diminished scrapie infectivity. Acid-induced transitions were reversible, whereas alkali produced an irreversible transition centered at pH 10 under conditions that diminished infectivity. Whether PrPSc synthesis involves a transition in the secondary structure of one or more domains of the cellular prion protein from alpha-helical, random coil, or turn into beta-sheet remains to be established. PMID- 8419913 TI - Brain weight and life-span in primate species. AB - In haplorhine primates (tarsiers, monkeys, apes, and humans), there is a significant correlation between brain weight and maximum life-span when the effect of body size is removed. There is also a significant correlation in haplorhine primates between brain weight and female age at first reproduction. For strepsirhine primates (lorises and lemurs), there are no significant correlations between brain weight and either life-span or female reproductive age when the effect of body size is removed. This lack of correlation in strepsirhine primates may be related to the fact that these primates are nocturnal and/or natives of the island of Madagascar, both of which conditions may reduce competition for resources and predation pressure. These findings suggest that in haplorhine primates the genetic systems controlling brain growth are linked to the systems governing the life cycle so that species with longer cycles have larger brains. When the effect of body weight is removed, leaf-eating haplorhines have significantly smaller brains and shorter lives than haplorhines with other diets. Harem-living haplorhines also have significantly smaller brains and shorter life-spans than troop-living haplorhines when the effect of body weight is removed. We also sought to test the rate-of-living hypothesis by determining whether primates with basal metabolic rates that are higher than would be expected for their body size have shorter maximum life-spans than would be expected for their body size. Metabolic rate is not correlated with life-span or female age at first reproduction when the effect of body size is removed. PMID- 8419914 TI - MtrB from Bacillus subtilis binds specifically to trp leader RNA in a tryptophan dependent manner. AB - MtrB regulates transcription attenuation of the Bacillus subtilis trp operon. We have shown that MtrB, either from B. subtilis or overexpressed in Escherichia coli, binds specifically to RNA from the leader region of the trp operon by a gel mobility-shift assay. This binding is tryptophan dependent. MtrB binds to a transcript terminated at the trp attenuator (-2 to +138) or a read-through transcript (-2 to +318). MtrB does not bind antisense trp leader RNA or single stranded trp leader DNA. These results support the model in which attenuation is controlled by tryptophan-activated MtrB influencing the secondary structure of the leader region transcript to form a terminator structure. PMID- 8419915 TI - The type 1 human immunodeficiency virus Tat binding protein is a transcriptional activator belonging to an additional family of evolutionarily conserved genes. AB - The type 1 human immunodeficiency virus Tat protein is a powerful transcriptional activator when bound to an RNA structure (TAR) present at the extreme 5' terminus of viral mRNA. Since transcriptional activation requires binding of Tat to RNA, it has been suggested that Tat enhances initiation or elongation through a direct interaction with cellular transcription factors. Here we show through protein fusion experiments that the previously identified cellular Tat binding protein, TBP-1, although unable to bind DNA, is a strong transcriptional activator when brought into proximity of several promoter elements. Transcriptional activity depends upon the integrity of at least two highly conserved domains: one resembling a nucleotide-binding motif and the other motif common to proteins with helicase activity. Our studies further reveal that TBP-1 represents one member of a large, highly conserved gene family that encodes proteins demonstrating strong amino acid conservation across species. Finally, we identified a second family member that, although 77% similar to TBP-1, does not activate transcription from the promoters examined. This finding, together with the observation that TBP-1 does not activate each promoter examined, suggests that this gene family may encode promoter-specific transcriptional activators. PMID- 8419916 TI - Cyclophilin-dependent stimulation of transcription by cyclosporin A. AB - Exposure to cyclosporin A (CspA) increased laccase (lac-1) transcript accumulation in the chestnut blight fungus Cryphonectria parasitica. This response was suppressed by compounds that interfere with calcium-dependent signal transduction and by the presence of a virulence-attenuating mycovirus. CspA stimulated the accumulation of mRNA from a nonhomologous reporter fused to the lac-1 promoter, indicating that the increased transcript levels resulted from an increase in promoter activity. Based on the current model for the regulation of lac-1 transcription, these results suggest that CspA interferes with a negative regulatory pathway that normally constrains lac-1 promoter activity. Significantly, CspA did not stimulate lac-1 transcription in mutant strains deficient in CspA binding activity, directly demonstrating a requirement for the interaction of CspA and cyclophilin in the modulation of lac-1 transcription. Our results establish that CspA treatment can stimulate gene transcription and that cyclophilin is the cellular receptor that mediates this activity. PMID- 8419917 TI - Serotonin 1B receptors in the developing somatosensory and visual cortices are located on thalamocortical axons. AB - Serotonin (5-HT)-immunoreactive axons are densely distributed in the primary visual and somatosensory cortices of rats, mice, and hamsters for the first 2 weeks of life, and a recent study from this laboratory has demonstrated that 5 HT1B receptors assume a pattern that exactly matches that of the serotoninergic axons. The differential distribution of these receptors is also transient. In the present study, we combined receptor binding autoradiography with neurochemical ablation of 5-HT axons or electrolytic lesions of the dorsal thalamus in an effort to determine the neural elements upon which the 5-HT1B receptors were located. Subcutaneous injections of the toxin 5,7-dihydroxytryptamine, made on the day of birth, totally eliminated the dense and patterned 5-HT innervation of the somatosensory and striate cortices of rats killed on postnatal day 8 but had no qualitative effect upon the distribution or density of 5-HT1B receptors in either of these cortical regions in animals killed at the same age. Conversely, electrolytic lesions of the dorsal thalamus made on postnatal day 6 resulted in a complete loss of the dense and patterned distribution of 5-HT1B receptors in rats killed on postnatal day 8. These results indicate that thalamocortical axons transiently express 5-HT1B receptors. PMID- 8419918 TI - Pooled-sampling makes high-resolution mapping practical with DNA markers. AB - A pooled-sample approach to the construction of high-resolution genetic maps is described. The strategy depends on the existence of an easily selectable target locus and the ability to produce large segregating populations. If these requirements are met, the pooled-sample mapping approach allows tightly linked markers (e.g., restriction fragment length polymorphisms) to be mapped relative to the target with a great economy of effort. The recombination fractions among loci can be estimated by the maximum likelihood method and a simple approximate estimator is derived. The order of loci is deduced using a Bayesian statistical framework to yield posterior probabilities for all possible orderings of a marker set. Optimal pooling strategies and the effects of misclassification of selected individuals are discussed and studied by computer simulation. The feasibility of this method is demonstrated by the high-resolution mapping of a region on chromosome 5 of tomato that contains a gene regulating fruit ripening. PMID- 8419919 TI - The role of zinc fingers in transcriptional activation by transcription factor IIIA. AB - We have described elsewhere a number of the properties of a set of mutant forms of Xenopus transcription factor IIIA (TFIIIA) containing single amino acid substitutions that result in the structural disruption of individual zinc finger domains. These "broken finger" proteins have now been analyzed with respect to their ability to support transcription of 5S rRNA genes in vitro. Disruption of any one of the first six zinc fingers of TFIIIA has no discernible effect on the activity of the protein in supporting 5S rRNA synthesis in standard in vitro transcription assays, despite the fact that some of these mutant proteins exhibit large decreases in their binding affinity for 5S rRNA genes in binary complexes. These results indicate that the activity of TFIIIA as a transcription factor can be largely independent of its equilibrium binding constant for the 5S rRNA gene in the absence of other components of the RNA polymerase III transcriptional apparatus. In fact, this finding is consistent with the known pathway and kinetics of assembly of 5S rRNA transcription complexes. In contrast to the results obtained with finger 1-6 mutants, analogous mutations in zinc fingers 7-9 of TFIIIA result in moderate to complete loss of transcriptional activity. We interpret these results to mean that the three C-terminal zinc fingers of TFIIIA are not only involved in binding to the internal control region of 5S rRNA genes but are also required, either directly or indirectly, for higher-order interactions that are important in transcription complex assembly, stability, or activity. PMID- 8419920 TI - The gene encoding the glutamate receptor subunit GluR5 is located on human chromosome 21q21.1-22.1 in the vicinity of the gene for familial amyotrophic lateral sclerosis. AB - Genomic clones of the human non-N-methyl-D-aspartate (non-NMDA) glutamate receptor subunit GluR5 were isolated by high-stringency screening of a cosmid library using the rat cDNA as a probe. The chromosomal localization of the human GluR5 gene has been established. Southern hybridization of DNA isolated from mapping panels of Chinese hamster-human hybrid cell lines and high-resolution in situ suppression hybridization localize the GluR5 gene to chromosome 21q21.1 22.1. This coincides with the localization of a mutant gene causing familial amyotrophic lateral sclerosis (ALS), as Siddique et al. established by linkage analyses [Siddique, T., Figlewicz, D. A., Pericak-Vance, M. A., Haines, J. L., Rouleau, G., Jeffers, A. J., Sapp, P., Hung, W. Y., Bebout, J., McKenna-Yasek, D., Deng, G., Horvitz, H. R., Gusella, J. F., Brown, R. H. & Roses, A. D. (1991) N. Engl. J. Med. 324, 1381-1384]. Convergent evidence from other investigators suggests that chronic pathologic activation of motor neurons via non-NMDA glutamate receptors might induce excitotoxic injury of motor neurons, culminating in ALS. Together with the demonstration that GluR5 transcripts are expressed in the ventral horn of the spinal cord, the region in which susceptible motor neurons reside, the chromosomal localization suggests that a mutated GluR5 gene may be responsible for the familial form of ALS. PMID- 8419921 TI - Selection of T lymphocytes bearing limited T-cell receptor beta chains in the response to a human pathogen. AB - Delayed-type hypersensitivity (DTH) is a classic measure of T-cell responsiveness to foreign antigen. To estimate the extent of the T-cell repertoire in the DTH response to a human pathogen, we measured T-cell receptor (TCR) beta-chain variable-region (V beta) gene usage in reversal reactions in leprosy. Reversal reactions represent naturally occurring DTH responses in leprosy, in which augmentation of T-cell responses to Mycobacterium leprae is concomitant with clearance of bacilli from lesions. T cells using the V beta 6-, V beta 12-, V beta 14-, and V beta 19-encoded TCRs were strikingly overrepresented in the lesions of patients as compared to blood and pre-DTH lesions from the same individuals. Furthermore, these data indicate a possible association between the predominant expression of a V beta gene segment in lesions and the major histocompatibility complex class II haplotype of the individual. V beta 6 was prominent in the lesions of four patients who were DR15, a marker of resistance in leprosy infection. Sequence analysis of V beta 6 TCRs showed frequent use of V beta 6.1 and J beta 2.7 gene segments and a conserved amino acid motif in the V-J junction in a reversal-reaction lesion, but not in blood from the same patient. The limited TCR repertoire expressed by the infiltrating T cells suggests that a limited set of antigens is recognized in the DTH response to a human pathogen. We suggest that the mechanism by which major histocompatibility complex haplotype influences DTH in this disease involves the presentation of specific peptides, with subsequent selection of specific TCRs followed by local oligoclonal expansion. PMID- 8419922 TI - Splanchnic metabolism of dietary arginine in relation to nitric oxide synthesis in normal adult man. AB - Urinary nitrate (NO3) is the stable end product of nitric oxide, which is formed, in turn, from a guanidino nitrogen of arginine. We have conducted two experiments, each in four healthy adult men receiving a low nitrate diet for 7-10 days, to investigate the in vivo conversion of arginine to nitrate. In the first study [guanidino-15N2, 5,5-2H2]arginine was given on day 7 via a primed continuous intravenous infusion for 8 h. In the second study, the labeled arginine was given for 8 h by the intragastric route on day 7 and by the intravenous route on day 10. Measurement of 15NO3 output in urine collected for 24 h beginning at the time of the arginine tracer infusion revealed a more extensive transfer of 15N when the arginine tracer was given intragastricly. From the comparative labeling of 15NO3 after administration of the tracer arginine via the intragastric and intravenous routes, we estimate that 16% +/- 2% of the daily production of nitrate arises from the metabolism of dietary arginine that is taken up during its "first pass" in the splanchnic region. Hence, nitric oxide production occurs, to a measurable extent, in this area in healthy subjects, raising the question as to how various pathophysiological states might alter the relations between exogenous and endogenous sources of arginine as precursors of NO. and the relative contributions made by various organs to whole body (NO.) NO3 formation. These results also raise important questions about the use of nitric oxide synthase inhibitors in animal and human studies. PMID- 8419923 TI - Endothelial cells produce a substance that inhibits contact activation of coagulation by blocking the activation of Hageman factor. AB - Human umbilical vein endothelial cells (HUVECs) produce a property that impairs the generation of coagulant and amidolytic activity initiated when normal human plasma is exposed to glass. This inhibitory property blocks the adsorption of Hageman factor (factor XII) to glass, thereby preventing the activation of Hageman factor, but does not impair the coagulant or amidolytic activity of already activated Hageman factor (factor XIIa). This property in HUVEC lysates could be neutralized by a purified preparation of Hageman factor but not by purified prekallikrein or high molecular mass kininogen. A partially purified inhibitory fraction from cell lysates exhibited a single homogeneous band in SDS/PAGE of approximately 22.5 kDa. Inhibitory activity was also found in concentrates of conditioned media from HUVECs, which also impaired the binding of Hageman factor to a surface; it may not be identical with that found in cell lysates. PMID- 8419924 TI - Cathepsin D gene is controlled by a mixed promoter, and estrogens stimulate only TATA-dependent transcription in breast cancer cells. AB - The cathepsin D (cath-D) gene, coding for a ubiquitous lysosomal aspartyl protease, is overexpressed in aggressive human breast cancers, and its transcription is induced by estrogens in hormone-responsive breast cancer cells. We have determined the structure and function of the proximal 5' upstream region of the human cath-D gene from MCF7 cells. We show that the promoter has a compound structure with features of both housekeeping genes (high G+C content and potential transcription factor Sp1 sites) and regulated genes (TATAA sequence). By RNase protection assay, we show that transcription is initiated at five major transcription sites (TSSI to -V) spanning 52 base pairs. In hormone-responsive breast cancer cells, estradiol increased by 6- to 10-fold the level of RNAs initiated at TSSI, which is located about 28 base pairs downstream from the TATA box. The specific regulation by estradiol of transcription starting at site I exclusively was confirmed by primer extension. Moreover, the same estradiol effect was observed in the ZR75-1 cell line and in MDA-MB231 estrogen-resistant breast cancer cells stably transfected with the estrogen receptor. Site-directed mutagenesis indicated that the TATA box is essential for initiation of cath-D gene transcription at TSSI. In breast cancer biopsy samples, high levels of TATA dependent transcription were correlated with overexpression of cath-D mRNA. We conclude that cath-D behaves, depending on the conditions, as a housekeeping gene with multiple start sites or as a hormone-regulated gene that can be controlled from its TATA box. PMID- 8419925 TI - Coordinated posttranscriptional control of gene expression by modular elements including Alu-like repetitive sequences. AB - We previously reported that in rat fibroblasts, accumulation of a set of mRNAs ("pIL genes") was modulated as a function of cell growth and transformation, at a posttranscriptional stage, and by a mechanism that depends on a short nucleotide sequence containing an ID repetitive element. In mouse fibroblasts, hybridization with rat pIL probes identified mRNAs with the same pattern of expression, which did not contain ID sequences but contained a different regulatory element, encompassing a repetitive sequence of the B1 family. Expression in mouse cells of a reporter beta-globin gene carrying this element inserted in its 3' noncoding region was growth- and transformation-dependent. The nucleotide sequences of two murine and of three rat pIL cDNAs showed clear similarities in the region immediately adjacent to the ID and B1 repeats. Both the repeat and the flanking sequence were required to confer on beta-globin constructs the pattern of expression characteristic of the pIL genes. The hypothesis is presented that repetitive sequences in the eukaryotic genome might be modular parts of complex regulatory elements ensuring the coordinated expression of various mRNA species. PMID- 8419926 TI - The phylogenetically conserved doublet tertiary interaction in domain III of the large subunit rRNA is crucial for ribosomal protein binding. AB - Previous phylogenetic analysis of rRNA sequences for covariant base changes has identified approximately 20 potential tertiary interactions. One of these is present in domain III of the large subunit rRNA and consists of two adjacent Watson-Crick base pairs that, in Saccharomyces cerevisiae 26S rRNA, connect positions 1523 and 1524 to positions 1611 and 1612. This interaction would strongly affect the structure of an evolutionarily highly conserved region that acts as the binding site for the early-assembling ribosomal proteins L25 and EL23 of S. cerevisiae and Escherichia coli, respectively. To assess the functional importance of this tertiary interaction, we determined the ability of synthetically prepared S. cerevisiae ribosomal protein L25 to associate in vitro with synthetic 26S rRNA fragments containing sequence variations at positions 1523 and 1524 and/or positions 1611 and 1612. Mutations that prevent the formation of both base pairs abolished L25 binding completely, whereas the introduction of compensatory mutations fully restored protein binding. Disruption of only the U1524.A1611 pair reduced L25 binding to approximately 30% of the value shown by the wild-type 26S rRNA fragment, whereas disruption of the G1523.C1612 base pair resulted in almost complete loss of protein binding. These results strongly support the existence and functional importance of the proposed doublet tertiary interaction in domain III of the large subunit rRNA. PMID- 8419927 TI - Cloning and characterization of the Salmonella typhimurium-specific chemoreceptor Tcp for taxis to citrate and from phenol. AB - Salmonella typhimurium shows an attractant response to citrate and a repellent response to phenol, and a chemoreceptor mediating these responses has been identified and named Tcp (taxis to citrate and away from phenol). Tcp is one of the methyl-accepting chemotaxis proteins that have a molecular mass of approximately 60 kDa estimated by SDS/PAGE, and its methylation level is increased by citrate and decreased by phenol. Tcp also mediates an attractant response to metal-citrate complexes. The complete nucleotide sequence of the tcp coding region has been determined. The deduced amino acid sequence of Tcp, consisting of 547-amino acid residues, is homologous with that of the aspartate chemoreceptor of S. typhimurium. Thus, Tcp is another member of the bacterial transmembrane chemoreceptor family. Because citrate is a good carbon source for S. typhimurium but is not a carbon source for the closely related species Escherichia coli and because citrate utilization is used as a key diagnostic character to distinguish these species, it is reasonable to assume that Tcp is specific to S. typhimurium. PMID- 8419928 TI - Apolipoprotein B mRNA editing is associated with UV crosslinking of proteins to the editing site. AB - Apolipoprotein (apo) B100 mRNA undergoes editing of C-6666 to a U residue, which generates a stop-translation codon and defines the carboxyl terminus of apoB48. To aid purification of the editing enzyme we have undertaken UV crosslinking of a 32P-labeled substrate for apoB mRNA editing in vitro to proteins in an enterocyte editing extract. Proteins of 60 (p60) and 43 (p43) kDa, prominent among crosslinking bands, were competed for by unlabeled substrate, but not by nonspecific RNA, and did not crosslink to antisense RNA. Editing in vitro and UV crosslinking were inhibited by NaCl and vanadyl ribonucleoside complexes and by chemical modification of sulfhydryl, imidazolium, and guanidinium groups on the protein. The editing activity copurified predominantly with p60. To define the binding site for p60 on the substrate RNA, a series of scanning and point mutant RNAs, previously used to define nucleotides 6671-6681 as essential for editing, were used in competition studies with wild-type substrate. Results demonstrated that p60 binding is centered on nucleotides 6671-6674. We suggest that p60 contains the RNA-recognition component of the apoB mRNA-editing enzyme. PMID- 8419929 TI - Sequence analysis of teleost retina-specific lactate dehydrogenase C: evolutionary implications for the vertebrate lactate dehydrogenase gene family. AB - At least two gene duplication events have led to the three lactate dehydrogenase (LDH; EC 1.1.1.27) isozymes (LDH-A, LDH-B, and LDH-C) of chordates. The prevailing model for the evolution of the LDH loci involves duplication of a primordial LDH locus near the origin of vertebrates, giving rise to Ldh-A and Ldh B. A third locus, designated Ldh-C, is expressed in the spermatocytes of mammals and a single family of birds and in the eye or liver tissues of teleost fishes. Ldh-C might have arisen independently in these taxa as duplications of either Ldh A or Ldh-B. Several authors have challenged this traditional hypothesis on the basis of amino acid sequence and immunological similarity of the three LDH isozymes. They suggest that the primordial LDH gene was duplicated to form Ldh-C and a locus that later gave rise to Ldh-A and Ldh-B. We have differentiated between these hypotheses by determining the cDNA sequence of the retina-specific LDH-C from a teleost, Fundulus heteroclitus. On the basis of amino acid sequence similarity, we conclude that the LDH-C isozymes in fish and mammals are not orthologous but derive from independent gene duplications. Furthermore, our phylogenetic analyses support previous hypotheses that teleost Ldh-C is derived from a duplication of the Ldh-B locus. PMID- 8419930 TI - Nitrilase in biosynthesis of the plant hormone indole-3-acetic acid from indole-3 acetonitrile: cloning of the Alcaligenes gene and site-directed mutagenesis of cysteine residues. AB - Indole-3-acetic acid is the major auxin in most plants. In Cruciferae, including Brassicaceae, indole-3-acetic acid is synthesized from indole-3-acetonitrile by nitrilase, after indole-3-acetonitrile is formed from tryptophan via indole-3 acetaldoxime or indole glycosinolates as the intermediate. We cloned and sequenced the gene for nitrilase (EC 3.5.5.1), which catalyzes the hydrolysis of indole-3-acetonitrile to indole-3-acetic acid, from Alcaligenes faecalis JM3. The amino acid sequence deduced from the nucleotide sequence of the nitrilase gene shows 34.7% identity with that of Klebsiella ozaenae nitrilase. A DNA clone containing the nitrilase gene expressed the active enzyme in Escherichia coli with excellent yield. Among five cysteine residues (Cys-40, Cys-115, Cys-162, Cys 163, and Cys-218) in the Alcaligenes nitrilase, only Cys-163 was conserved at the corresponding position in the Klebsiella nitrilase. Two mutant enzymes, in which Cys-162 and Cys-163 were replaced with Asn and Ala, respectively, were constructed by site-directed mutagenesis. A 35% increase of the specific activity and a large reduction of the Km for thiophene-2-acetonitrile (which was used as a standard substrate for the nitrilase) were observed in the Cys-162-->Asn mutant enzyme. The Cys-163-->Ala mutation resulted in complete loss of nitrilase activity, clearly indicating that Cys-163 is crucial for the activity and Cys-162 could not provide the catalytic function of Cys-163. PMID- 8419931 TI - Tyrosine phosphorylation is a mandatory proximal step in radiation-induced activation of the protein kinase C signaling pathway in human B-lymphocyte precursors. AB - Ionizing radiation triggers a signal in human B-lymphocyte precursors that is intimately linked to an active protein-tyrosine kinase regulatory pathway. We show that in B-lymphocyte precursors, irradiation with gamma-rays leads to (i) stimulation of phosphatidylinositol turnover; (ii) downstream activation by covalent modification of multiple serine-specific protein kinases, including protein kinase C; and (iii) activation of nuclear factor kappa B. All of the radiation-induced signals were effectively prevented by the protein-tyrosine kinase inhibitors genistein and herbimycin A. Thus, tyrosine phosphorylation is an important and perhaps mandatory proximal step in the activation of the protein kinase C signaling cascade in human B-lymphocyte precursors. Our report expands current knowledge of the radiation-induced signaling cascade by clarifying the chronological sequence of biochemical events that follow irradiation. PMID- 8419932 TI - Characterization of a G-protein-regulated outward K+ current in mesophyll cells of vicia faba L. AB - Whole-cell voltage-dependent currents in isolated mesophyll protoplasts of Vicia faba were investigated by patch-clamp techniques. With 104 mM K+ in the cytosol and 13 mM K+ in the external solution, depolarization of the plasma membrane from -47 mV to potentials between -15 and +85 mV activated a voltage- and time dependent outward current (Iout). The average magnitude of Iout at +85 mV was 28.5 +/- 3.3 pA.pF-1. No inward voltage-dependent current was observed upon hyperpolarization of the plasma membrane from -55 mV to potentials as negative as -175 mV. Time-activated outward current was blocked by Ba2+ (1 mM BaCl2) and was not observed when K+ was eliminated from the external and internal solutions, indicating that this outward current was carried primarily by K+ ions. The voltage dependency of outward K+ current revealed a possible mechanism for K+ efflux from mesophyll cells. A GDP analogue guanosine 5'-[beta-thio]diphosphate (500 microM) significantly enhanced outward K+ current. The outward K+ current was inhibited by the GTP analogue guanosine 5'-[gamma-thio]triphosphate (500 microM) and by an increase in cytoplasmic free Ca2+ concentrations. Cholera toxin, which ADP-ribosylates guanine nucleotide-binding regulatory proteins, also inhibited outward K+ current. These findings illustrate the presence in mesophyll cells of outward-rectifying K+ channels that are regulated by GTP-binding proteins and calcium. PMID- 8419933 TI - Characterization of cis-acting elements regulating transcription of the human DF3 breast carcinoma-associated antigen (MUC1) gene. AB - The present studies have examined the sequences responsible for regulating transcription of the human DF3 breast carcinoma-associated antigen (MUC1) gene. A region 1656 base pairs upstream to the DF3 transcription initiation site was fused to the chloramphenicol acetyltransferase gene. Transient expression assays using a series of deleted constructs demonstrated that the region from position 618 contains the regulatory sequences necessary for DF3 transcription in human MCF-7 breast cancer cells. Further analysis with internal deletion vectors and heterologous promoter constructs indicated the involvement of cis-acting elements in the fragment extending from positions -598 to -485. By gel retardation and DNA footprinting, we have identified a protein in MCF-7 cells that recognizes sequences between positions -505 and -485. The results of Southwestern studies demonstrate that this protein has an apparent molecular mass of 45 kDa. Taken together, these results suggest that DF3 gene transcription is regulated by a previously undescribed transacting factor. PMID- 8419934 TI - High-frequency genetic reversion mediated by a DNA duplication: the mouse pink eyed unstable mutation. AB - The mouse pink-eyed unstable (p(un)) mutation, affecting coat color, exhibits one of the highest reported reversion frequencies of any mammalian mutation and is associated with a duplication of genomic DNA at the p locus. In this study, genomic clones containing the boundaries of the p(un) duplication were isolated and characterized. The structure of these sequences and their wild-type and revertant counterparts were analyzed by restriction mapping, PCR product analysis, DNA sequence analysis, and pulsed-field gel electrophoresis. DNA from p(un) was distinguished from wild-type and revertant DNA by a head-to-tail tandem duplication of approximately 70 kilobases. No differences were detected between revertant and wild-type DNAs. Thus, the reversion in phenotype of p(un) mice is coupled with the loss of one copy of an approximately 70-kilobase duplicated segment. Testable models are presented to account for p(un) reversion. PMID- 8419935 TI - Blood-brain barrier penetration abolished by N-methyl quaternization of nicotine. AB - The present study determined the effect of organically quaternizing either of the two tertiary nitrogen sites of nicotine to assess the in vivo effects of the permanently ionized states of the synthesized N-[14C]methylnicotines on brain uptake in rat after intracarotid injection. Male Sprague-Dawley rats were used to measure the brain uptake index (BUI) by single-pass clearance in brain after rapid injection at pH 7.4 into the left common carotid artery (expressed as a percentage) relative to simultaneously injected 3HOH. The BUI of [14C]mannitol, a control for the method background, was measured to be 2.6 +/- 0.6. At physiological pH, in striking contrast to the [pyrrolidine-2-14C]nicotine BUI of 120 +/- 3, the N-[14C]-methylnicotines had a BUI of 3.0 +/- 0.6, which was not significantly different from the method background and which indicated abolition of blood-brain barrier penetration of nicotine with the sensitivity of the BUI method. PMID- 8419936 TI - Glutathione ester delays the onset of scurvy in ascorbate-deficient guinea pigs. AB - Previous studies showed that administration of ascorbate to glutathione (GSH) deficient newborn rats and guinea pigs prevented toxicity and mortality and led to increased tissue and mitochondrial GSH levels; ascorbate thus spares GSH. In the present work, we tried to answer the converse question: Does administration of GSH spare ascorbate? Because administered GSH is not well transported into most cells, we gave GSH monoethyl ester (which is readily transported and converted into GSH intracellularly) to guinea pigs fed an ascorbate-deficient diet. We found that treatment with GSH ester significantly delays appearance of the signs of scurvy and that this treatment spares ascorbate; thus, the decrease of tissue levels of ascorbate was delayed. The findings support the conclusions that (i) GSH is essential for the physiological function of ascorbate because it is required in vivo for reduction of dehydroascorbate and (ii) there is metabolic redundancy and overlap of the functions of these antioxidants. The sparing effect of GSH in scurvy may be mediated through an increase in the reduction of dehydroascorbate (which would otherwise be degraded) and to antioxidant effects of GSH that are also produced by ascorbate. Other studies indicate that GSH deficiency in adult mice stimulates ascorbate synthesis in liver. During this work we found that administration of GSH itself is highly toxic to ascorbate deficient guinea pigs when given in divided i.p. doses totaling 3.75 mmol/kg daily. PMID- 8419937 TI - Spontaneous and radiation-induced renal tumors in the Eker rat model of dominantly inherited cancer. AB - Hereditary renal carcinoma (RC) in the rat, originally reported by R. Eker in 1954, is an example of a Mendelian dominant predisposition to a specific cancer in an experimental animal. At the histologic level, RCs develop through multiple stages from early preneoplastic lesions (e.g., atypical tubules) to adenomas in virtually all heterozygotes by the age of 1 year. The homozygous mutant condition is lethal at approximately 10 days of fetal life. Ionizing radiation induces additional tumors in a linear dose-response relationship, suggesting that in heterozygotes two events (one inherited, one somatic) are necessary to produce tumors, and that the predisposing gene is a tumor suppressor gene. No genetic linkage has yet been found between the Eker mutation and rat DNA sequences homologous to those in human chromosome 3p, the presumed site of the putative tumor suppressor gene responsible for human RC. Nonrandom loss of rat chromosome 5 in RC-derived cell lines is sometimes associated with homozygous deletion of the interferon gene loci at rat chromosome bands 5q31-q33. Since this locus is not linked with the predisposing inherited gene in the Eker rat, it probably represents a second tumor suppressor gene involved in tumor progression. PMID- 8419938 TI - Soluble CD4 enhances the efficacy of immunotoxins directed against gp41 of the human immunodeficiency virus. AB - Monoclonal antibodies specific for the gp120 or gp41 portions of the human immunodeficiency virus (HIV) envelope protein gp160 were conjugated to ricin A chain, and their immunotoxic activities against HIV-infected cells were evaluated in the presence or absence of soluble CD4 (sCD4). Immunotoxin activity was measured in vitro as cytotoxicity and inhibition of secretion of infectious HIV. The efficacy of anti-gp41 immunotoxins was enhanced at least 30-fold in the presence of sCD4. This effect was specific for HIV-infected cells, but not for uninfected cells, and was seen at concentrations of sCD4 as low as 0.1 micrograms/ml. Anti-gp120 immunotoxins were marginally inhibited at higher concentrations of sCD4. Flow cytometry analyses showed that sCD4 increased the expression of gp41 on the surface of infected cells and increased internalization of gp120 and gp41. These data suggest that sCD4 alters the cellular trafficking of HIV envelope proteins. These findings also have important implications for the therapeutic use of anti-HIV immunotoxins and may be generalizable to other immunotoxins as well. PMID- 8419939 TI - Crystal structure of phospholipase A2 from Indian cobra reveals a trimeric association. AB - Phospholipase A2 (PLA2) from Indian cobra venom (Naja naja naja) was crystallized from ethanol in space group P4(3)2(1)2 in the presence of Ca2+. The x-ray crystal structure was determined to 2.3-A resolution by molecular replacement techniques using a theoretical model constructed from homologous segments of the bovine pancreatic, porcine pancreatic, and rattlesnake venom crystal structures. The structure was refined to an R value of 0.174 for 17,542 reflections between 6.0- and 2.3-A resolution (F > 2 sigma), including 148 water molecules. The 119-amino acid enzyme has an overall architecture strikingly similar to the other known PLA2 structures with regions implicated in catalysis showing the greatest structural conservation. Unexpectedly, three monomers were found to occupy the asymmetric unit and are oriented with their catalytic sites facing the pseudo threefold axis with approximately 15% of the solvent accessible surface of each monomer buried in trimer contacts. The majority of the interactions at the subunit interfaces are made by residues unique to PLA2 sequences from cobra and krait venoms. The possible relevance of this unique trimeric structure is considered. PMID- 8419940 TI - Photolabeling evidence for calcium-induced conformational changes at the ATP binding site of scallop myosin. AB - A change in the conformation of the active site of scallop myosin under the influence of regulatory amounts of Ca2+ has been identified by use of the ADP photoaffinity analog 2-[(4-azido-2-nitrophenyl)amino]ethyl diphosphate (NANDP). NANDP, trapped at the active site with Mn2+ and vanadate, photolabeled preferentially Arg-128 of the heavy chain in the absence of added Mg2+ and Ca2+ [Kerwin, B. & Yount, R. (1992) Bioconjugate Chem. 3, 328-336]. However, addition of 2 mM Mg2+ and regulatory amounts of Ca2+ (0.01-1 microM) shifted the predominant labeling to Cys-198 of the heavy chain in a Ca(2+)-dependent manner. This Ca(2+)-dependent change in the photolabeling pattern was absent when the regulatory light chains were removed or when the unregulated head (subfragment 1) was examined under similar conditions. These results demonstrate that both Arg 128 and Cys-198 are part of the purine binding site which undergoes a conformational change in response to Ca2+ binding to the regulatory domain. PMID- 8419941 TI - Analysis of human immunodeficiency virus-infected tissues by amplification and in situ hybridization reveals latent and permissive infections at single-cell resolution. AB - Latent and productive viral infections are at the extremes of the spectrum of virus-cell interactions that are thought to play a major role in the ability of such important human pathogens as human immunodeficiency virus (HIV) to elude host defenses and cause disease. The recent development of PCR-based methods to amplify target sequences in individual cells in routinely fixed tissues affords opportunities to directly examine the subtle and covert virus-cell relationships at the latent end of the spectrum that are inaccessible to analysis by conventional in situ hybridization techniques. We have now used PCR in situ with in situ hybridization to document latent and permissive HIV infection in routinely fixed and paraffin-embedded tissue. In one of the first specimens we examined, a tumor biopsy from an HIV-infected individual, we found many of the lymphocytes and lymphocytes infiltrating the tumor had HIV DNA that was detectable only by PCR in situ. The fraction of positive cells varied regionally, but there were foci where most of the cells contained HIV DNA. Most of these lymphocytes and macrophages are latently infected, as we could detect HIV RNA in fewer than one in a thousand of these cells. We also detected HIV RNA, surprisingly, in 6% of the tumor cells, where the number of copies of viral RNA per cell was equivalent to productively infected cell lines. The alternative states of HIV-gene expression and high local concentration of latently infected lymphocytes and monocytes revealed by these studies conceptually supports models of lentiviral pathogenesis that attribute persistence to the reservoir of latently infected cells and disease to the consequences of viral-gene expression in this population. The magnitude of infection of lymphocytes documented in this report is also consistent with the emerging view that HIV infection per se could contribute substantially to depletion of immune cells in AIDS. PMID- 8419942 TI - Palmitoylation of bovine opsin and its cysteine mutants in COS cells. AB - Previously, bovine rhodopsin has been shown to be palmitoylated at cysteine residues 322 and 323. Here we report on palmitoylation of bovine opsin in COS-1 cells following expression of the synthetic wild-type opsin gene and several of its cysteine mutants in the presence of [3H]palmitic acid. Two moles of palmitic acid are introduced per wild-type opsin molecule in thioester linkages. Palmitoylation is abolished when both Cys-322 and Cys-323 are replaced by serine residues. Replacement of Cys-322 by serine prevents palmitoylation at Cys-323, whereas replacement of the latter with serine allows palmitoylation at Cys-322. Opsin mutants that evidently do not contain a Cys-110/Cys-187 disulfide bond and presumably remain in the endoplasmic reticulum are not palmitoylated. Replacement of Cys-140 or Cys-185 reduces the extent of palmitoylation of the opsin. Lack of palmitoylation at Cys-322 and/or Cys-323 does not affect 11-cis-retinal binding, absorption maximum or extinction coefficient of the chromophore, the bleaching behavior of the chromophore, or the light-dependent binding and activation of transducin. Mutants containing serine substitutions at Cys-140 or Cys-323 showed reduced light-dependent phosphorylation by rhodopsin kinase. PMID- 8419943 TI - Autoxidation of lipids and antioxidation by alpha-tocopherol and ubiquinol in homogeneous solution and in aqueous dispersions of lipids: unrecognized consequences of lipid particle size as exemplified by oxidation of human low density lipoprotein. AB - Recent studies on the initial stages in oxidation of low density lipoprotein (LDL) have revealed certain previously unrecognized similarities to emulsion polymerization and some quite unexpected features including the following: (i) ascorbate is an extremely effective antioxidant for LDL containing alpha tocopherol (alpha-TOH); (ii) in the presence of alpha-TOH and in the absence of both ascorbate and ubiquinol 10 (Q10H2), oxidation of LDL occurs via a free radical chain; (iii) Q10H2 is a much better antioxidant for LDL than alpha-TOH, although the reverse is true in homogeneous systems. We show here that these problems can be solved on the basis of three simple hypothesis, each of which is based on known chemistry: (i) alpha-TOH in LDL can be regenerated from its radical, alpha-TO., by ascorbate; (ii) in the absence of ascorbate and Q10H2, the alpha-TOH in LDL acts as a chain-transfer agent rather than as a radical trap; (iii) Q10H2 is a much more effective chain-breaking antioxidant than alpha-TOH in LDL because the semiquinone radical Q10H. exports its radical character from the LDL into the aqueous phase. Our conclusions imply that the search for better antiatherosclerotic drugs might profitably focus on antioxidants capable of exporting radicals from LDL particles or otherwise increasing the traffic of radicals between particles. PMID- 8419944 TI - O2A progenitor cells transplanted into the neonatal rat brain develop into oligodendrocytes but not astrocytes. AB - The differentiation of the bipotential O2A progenitor cell into an oligodendrocyte or a type 2 astrocyte has been well documented in cell cultures of various regions of the central nervous system. The appropriate tools to prove its existence in vivo have been lacking. We report on an in vitro-in vivo approach that combines stable labeling of an enriched population of cultured O2A progenitors by the fluorescent dye fast blue, followed by their transplantation into neonatal rat brains, which allowed us to study the influence of the brain microenvironment on their lineage decision. The grafted cells survived well and 21 days after grafting nearly all were positive for the oligodendroglial marker galactocerebroside. Surprisingly, the fast blue-positive grafted cells did not stain for the astroglial marker glial fibrillary acidic protein. These results indicate that the O2A progenitor's plasticity is restricted by the in vivo environment, resulting in the developmental exclusion of the type 2 astrocyte initially described in vitro. PMID- 8419945 TI - Activation of glucose transport by a natural mutation in the human insulin receptor. AB - Naturally occurring mutations in the insulin receptor gene cause heritable severe insulin resistance. These mutations usually impair insulin receptor signaling in cells cultured from affected individuals. However, fibroblasts cultured from a patient with intrauterine growth restriction and severe insulin resistance (leprechaun Atl-1) had normal amounts of insulin receptor protein and defective insulin binding but constitutive activation of insulin-receptor autophosphorylation and kinase activity and of glucose transport. In the same fibroblasts, growth was impaired. Homozygosity for a mutation in the insulin receptor gene was suspected, since he inherited identical DNA haplotypes for this gene from both related parents. Here we report that the proband was homozygous and both parents were heterozygous for a point mutation in the insulin receptor gene converting the Arg86 codon (CGA) to Pro (CCA) (R86P). The R86P substitution is contiguous to the hydrophobic beta-sheet of the receptor alpha subunit implicated by DeMeyts et al. [DeMeyts, P., Gu, J.-L., Shymko, R. M., Kaplan, B. E., Bell, G. I. & Whittaker, J. (1990) Mol. Endocrinol. 4, 409-416] in the binding of aromatic residues of the insulin molecule. The R86P mutant insulin receptor cDNA was inserted into a plasmid under control of a simian virus 40 promoter and transfected into Chinese hamster ovary (CHO) cells. In contrast with fibroblasts from patient Atl-1, which had normal insulin receptor processing, CHO cells stably transfected with the R86P mutant cDNA (CHO-R86P) had altered posttranslational processing. The R86P mutant receptor failed to bind insulin but caused a significant increase in basal glucose transport in CHO cells. As in fibroblasts cultured from the patient, the R86P mutant insulin receptor did not stimulate growth in transfected CHO cells. These results suggest that the R86P mutation in the insulin receptor activates glucose transport without promoting cell growth and that distinct cell types process this mutant insulin receptor differently. PMID- 8419946 TI - Zaragozic acids: a family of fungal metabolites that are picomolar competitive inhibitors of squalene synthase. AB - Three closely related fungal metabolites, zaragozic acids A, B, and C, that are potent inhibitors of squalene synthase have been isolated and characterized. Zaragozic acids A, B, and C were produced from an unidentified sterile fungal culture, Sporormiella intermedia, and Leptodontium elatius, respectively. The structures of the zaragozic acids and their trimethyl esters were determined by a combination of physical and chemical techniques. The zaragozic acids are characterized by a novel 2,8-dioxobicyclo[3.2.1]octane-4,6,7- trihydroxyl-3,4,5 tricarboxylic acid core and differ from each other in the structures of the 6 acyl and 1-alkyl side chains. They were found to be potent competitive inhibitors of rat liver squalene synthase with apparent Ki values of 78 pM, 29 pM, and 45 pM, respectively. They inhibited cholesterol synthesis in Hep G2 cells, and zaragozic acid A was an inhibitor of acute hepatic cholesterol synthesis in the mouse (50% inhibitory dose of 200 micrograms/kg of body weight). Inhibition of squalene synthase in cells and in vivo was accompanied by an accumulation of label from [3H]mevalonate into farnesyl diphosphate, farnesol, and organic acids. These data indicate that the zaragozic acids are a previously unreported class of therapeutic agents with potential for the treatment of hypercholesterolemia. PMID- 8419947 TI - Steroids and their conjugates in the mammalian brain. AB - Five steroids--3 beta-hydroxypregn-5-en-20-one (pregnenolone; P), 3 beta-hydroxy 5 alpha-pregnan-20-one (3 beta-AP), 3 alpha-hydroxy-5 alpha-pregnan-20-one (3 alpha-AP), 3 beta-hydroxyandrost-5-en-17-one (dehydroepiandrosterone; D), and 3 beta-hydroxy-5 alpha-androstan-17-one (EpiA)--were extracted from the brains of adult male rats, rabbits, and dogs. The steroids exist in this organ as unconjugated compounds and as sulfates, lipoidal esters, and sulfolipids. The techniques for separating these four classes of steroids from each other and for separating the five steroids from each other are described. In all cases, the steroids were identified by their retention time (Rt) on HPLC, their Rt by gas chromatography, and by selected ion monitoring of their mass spectra. The latter were also used for quantification. In their reaction toward organic bases, the sulfolipid conjugates resemble previously described sulfolipids of cholesterol and sitosterol. These conjugates are relatively abundant in brain, particularly those of P and D, and this suggests that, in the search for the physiological significance of these brain constituents, these conjugates warrant attention. PMID- 8419948 TI - Hypervariable-control-region sequences reveal global population structuring in a long-distance migrant shorebird, the Dunlin (Calidris alpina). AB - Hypervariable segments of the control region of mtDNA as well as part of the cytochrome b gene of Dunlins were amplified with PCR and sequenced directly. The 910 base pairs (bp) obtained for each of 73 individuals complete another of the few sequencing studies that examine the global range of a vertebrate species. A total of 35 types of mtDNA were detected, 33 of which were defined by the hypervariable-control-region segments. Thirty of the latter were specific to populations of different geographic origin in the circumpolar breeding range of the species. The remaining three types indicate dispersal between populations in southern Norway and Siberia, but female-mediated flow of mtDNA apparently is too low to overcome the effects of high mutation rates of the control-region sequences, as well as population subdivision associated with historical range disjunctions. A genealogical tree relating the types grouped them into five populations: Alaska, West Coast of North America, Gulf of Mexico, western Europe, and the Taymyr Peninsula. The Dunlin is thus highly structured geographically, with measures of mutational divergence approaching 1.0 for fixation of alternative types in different populations. High diversity of types within populations as well as moderate long-term effective population sizes argue against severe population bottlenecks in promoting this differentiation. Instead, population fragmentation in Pleistocene refuges is the most plausible mechanism of mtDNA differentiation but at a much earlier time scale than suggested previously with morphometric data. PMID- 8419949 TI - People need rights over their own death. PMID- 8419950 TI - Paving the way to acceptance. Psychological adaptation to death and dying in cancer. AB - Dying people often experience a range of strong emotions--denial, anger, bargaining and depression--before coming to accept the near prospect of their death. By offering support and being prepared to listen, nurses can help patients and their families express their feelings and possibly proceed onto acceptance. PMID- 8419951 TI - Treatment we can all believe in: pain and its management in later life. PMID- 8419952 TI - A checklist to meet long-term needs. Continence assessment of neuroscience patients. AB - The continence-related problems which affect neuroscience patients are often overlooked. Using a simple checklist can ensure adequate assessment is undertaken to facilitate effective long-term management of this problem. PMID- 8419953 TI - A positive coping strategy. Humour in the oncology setting. AB - Humour is an integral part of most positive human relationships, and does much to bring people closer together. Used sensitively and tactfully, it has much to offer patients and nurses in an oncology setting. PMID- 8419954 TI - A systematic approach with lasting benefits: designing and implementing a wound assessment chart. AB - Applying a systematic approach to wound assessment is known to have long-term advantages. This study describes how two infection control nurses designed and implemented a wound assessment chart, which was intended to produce permanent changes in practice. PMID- 8419955 TI - A system to meet nursing needs. The project nurse's role in computer procurement. AB - Computers are an essential component of most quality initiatives and in resource management. The project nurse role has been devised to ensure staff are consulted in the procurement process, and that a computer system is chosen which meets their needs. PMID- 8419956 TI - The right time to give up. Advising women on smoking in pregnancy. AB - Smoking in pregnancy can be harmful to the fetus, and it is a good idea for women to stop, or at least reduce, their cigarette consumption--before they conceive if possible. Nurses are well-placed to offer women advice on this. PMID- 8419957 TI - Smoking and pregnancy. PMID- 8419958 TI - A requirement not to be overlooked. Nutritional aspects of respiratory disease. AB - For many people with severe respiratory disease malnutrition can become a major problem. By considering the practical and physical difficulties caused by the respiratory symptoms and more accurately assessing nutritional needs, an inadequate dietary intake can be overcome, leading to a better nutritional status. PMID- 8419959 TI - Assessing contraceptive needs during the menopause. PMID- 8419960 TI - Support to show they are not alone. Stress in district nursing. AB - The growing emphasis on community care and the comparative isolation of its role is making district nursing increasingly stressful. Support from colleagues and management can help relieve the pressure, and will ultimately benefit patient care. PMID- 8419961 TI - An education in idealism or reality? PMID- 8419962 TI - Interstitial-lymphatic mechanisms in the control of extracellular fluid volume. AB - While the study of the physiochemical composition and structure of the interstitium on a molecular level is a large and important field in itself, the present review centered mainly on the functional consequences for the control of extracellular fluid volume. As pointed out in section I, a biological monitoring system for the total extracellular volume seems very unlikely because a major part of that volume is made up of multiple, separate, and functionally heterogeneous interstitial compartments. Even less likely is a selective volume control of each of these compartments by the nervous system. Instead, as shown by many studies cited in this review, a local autoregulation of interstitial volume is provided by automatic adjustment of the transcapillary Starling forces and lymph flow. Local vascular control of capillary pressure and surface area, of special importance in orthostasis, has been discussed in several recent reviews and was mentioned only briefly in this article. The gel-like consistency of the interstitium is attributed to glycosaminoglycans, in soft connective tissues mainly hyaluronan. However, the concept of a gel phase and a free fluid phase now seems to be replaced by the quantitatively more well-defined distribution spaces for glycosaminoglycans and plasma protein, apparently in osmotic equilibrium with each other. The protein-excluded space, determined mainly by the content of glycosaminoglycans and collagen, has been measured in vivo in many tissues, and the effect of exclusion on the oncotic buffering has been clarified. The effect of protein charge on its excluded volume and on interstitial hydraulic conductivity has been studied only in lungs and is only partly clarified. Of unknown functional importance is also the recent finding of a free interstitial hyaluronan pool with relatively rapid removal by lymph. The postulated preferential channels from capillaries to lymphatics have received little direct support. Thus the variation of plasma-to-lymph passage times for proteins may probably be ascribed to heterogeneity with respect to path length, linear velocity, and distribution volumes. Techniques for measuring interstitial fluid pressure have been refined and reevaluated, approaching some concensus on slightly negative control pressures in soft connective tissues (0 to -4 mmHg), zero, or slightly positive pressure in other tissues. Interstitial pressure volume curves have been recorded in several tissues, and progress has been made in clarifying the dependency of interstitial compliance on glycosaminoglycan osmotic pressure, collagen, and microfibrils.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419963 TI - Taurine in development. PMID- 8419964 TI - Cellular and molecular mechanisms of aging. PMID- 8419965 TI - Biology and biochemistry of proteinases in tumor invasion. PMID- 8419966 TI - The biochemical basis of zinc physiology. PMID- 8419967 TI - Preoperative imaging anatomy of nasal-ethmoid complex for functional endoscopic sinus surgery. AB - Endoscopic sinus surgery has become an increasingly popular surgical procedure. Functional endoscopic sinus surgery is based on the hypothesis that the ostiomeatal complex is the key area in the pathogenesis of chronic sinusitis. This article discusses the concept of functional endoscopic sinus surgery, the anatomy of the ostiomeatal complex, and the imaging anatomy of the ostiomeatal complex. PMID- 8419968 TI - Imaging of cysts and odontogenic tumors of the jaw. Definition and classification. AB - For the evaluation of jaw cysts and odontogenic tumors, the radiologic examinations include conventional films, computed tomographic (CT) scans, and magnetic resonance (MR) images. The indications for these radiologic methods are discussed along with the radiologic findings for the various cysts and tumors described. The differential diagnosis of these lesions is also included in the discussion. PMID- 8419969 TI - Fibro-osseous lesions of craniofacial bones. The role of imaging. AB - Fibro-osseous lesions of the craniofacial structures are a group of pathologic conditions that are somewhat difficult to classify. Fibro-osseous lesions of the jaw may be divided into two categories: fibrous dysplasia, a developmental lesion characterized by the development of swelling, which consists of proliferating fibrous tissue that replaces normal bone; and those that have been postulated to originate from the periodontal ligament. This article describes the imaging characteristics of fibrous dysplasia and other fibro-osseous lesions of oral bones, including ossifying fibroma, periapical cemental dysplasia, cemento ossifying fibroma, and florid osseous dysplasia. PMID- 8419970 TI - Diagnosis of pathology of the temporomandibular joint. Clinical and imaging perspectives. AB - The pathologic conditions that involve the TMJ are similar to those conditions that involve other joints in the body. Therefore, many of the radiologic characteristics of these diseases are also similar. Nevertheless, because the articular surface of the condyle is an important site of mandibular growth, leading to secondary facial asymmetries when damaged in young patients, this possibility must be considered when evaluating the radiographic changes. It also is important to understand that functionally the TMJs act as a single unit and that any alteration in function caused by a pathologic process on one side can lead to disturbed function on the opposite side. Such dysfunction can cause degenerative changes that must be distinguished from the primary disease. Finally, some of the conditions that involve the TMJ can produce rather similar radiographic changes. In these instances, an understanding of the associated clinical findings can be helpful in making a more accurate radiographic diagnosis. PMID- 8419971 TI - Imaging of internal derangements and synovial chondromatosis of the temporomandibular joint. AB - Magnetic resonance (MR) imaging of internal derangements and synovial chondromatosis is described. The MR findings of fluid collections, increased vascularity, partial disc displacement, and changes in disc morphology and length are among those factors that influence surgical execution. Fast imaging is particularly useful in delineating fluid and vascularity changes. Synovial chondromatosis is an entity characterized by large intra-articular fluid collections and is thus ideally evaluated using MR imaging. PMID- 8419972 TI - The role of diagnostic imaging in dental implantology. AB - Dental implants are titanium cylinders that are surgically implanted into the jaw to allow fixation of a permanent dental prosthesis. Dental computed tomographic (CT) software programs that display multiple axial, cross-sectional and panoramic images of the jaw have been developed to assess these patients preoperatively. The development and use of these programs, the implant surgical procedure, and related dental anatomy and oral pathology are discussed. PMID- 8419973 TI - Applications of three-dimensional CT imaging in head and neck pathology. AB - The advantages of three-dimensional (3-D) imaging in the diagnosis of developmental and posttraumatic craniofacial abnormalities is well established. A brief review of this role of 3-D imaging is presented, followed by a discussion of the use of 3-D imaging in various head and neck disorders. PMID- 8419974 TI - Advanced imaging of osseous maxillary clefts. AB - A computed tomographic (CT) technique to establish precise two-dimensional (2-D) and three-dimensional (3-D) images of the osseous defects of cleft palates is presented and illustrated by two case studies. Prospective soft tissue algorithms and bone detail imaging was made possible by a retrospective program, a specific software program and vertical reformatting technique leading to 3-D image reconstruction. The two cases illustrate the flexibility of the CT program in accurately providing morphometric and bone density data on the location and size of the osseous defects involved in the cleft. Not every cleft palate patient is a candidate for the procedures outlined; however, the diagnosis of and treatment planning for patients presenting with bilateral or extensive osseous clefting can be more accurate. PMID- 8419975 TI - Clinical aspects of imaging in maxillofacial trauma. AB - Imaging of maxillofacial injuries serves as the principal means of qualifying the clinical diagnosis. Imaging assists in treatment planning of comprehensive surgical repair. The use of more invasive means of providing stable fixation of facial fractures has increased the demands made on fracture imaging. Clinical decision making depends on a thorough understanding of the clinical findings in an injury and its anatomic features as delineated on imaging. PMID- 8419976 TI - Complications of endoscopic sinus surgery. The role of the radiologist in prevention. AB - Intranasal surgery via the endoscope has become an extremely popular technique for refractory rhinosinusitis, but like all procedures it carries a risk of complications. Mosher in 1912 stated that intranasal ethmoidectomy is "the blindest and most dangerous (operation) in all surgery." Complications of a surgical procedure are among the most devastating problems in the practice of medicine. A thorough knowledge on the part of the radiologist of what the otolaryngologist is attempting to do can only enhance patient care. All screening CT scans should be approached as if they are preoperative examinations. It is not enough for the radiologist to merely describe air-fluid levels or mucoperiosteal thickening. Instead, the radiologist is an important member of the health care team, and an active role should be assumed to describe anatomic variants that may place the patient at increased surgical risk and to assist the surgeon in the appropriate evaluation of the patient who has a surgical complication. PMID- 8419977 TI - Preoperative and postoperative imaging evaluation of patients with maxillofacial deformities. AB - This article discusses the various imaging techniques that are most helpful in the diagnosis, treatment planning, and outcome assessment of patients with maxillofacial deformities. Specific anatomic inter-relationships of which the radiologist should be aware are discussed to aid in communication with the treating physician. Imaging characteristics of specific maxillofacial deformities and common syndromes are presented. PMID- 8419978 TI - Chronic inflammatory sinonasal diseases including fungal infections. The role of imaging. AB - The computed tomographic (CT) and magnetic resonance (MR) imaging techniques used for evaluating the paranasal sinuses and nasal fossae are reviewed. A summary of the physics concepts relating to the MR signal intensities observed with sinonasal secretions is given, as is a review of normal secretions and the pathophysiologic changes in the secretions that occur with chronic infections. The imaging findings of sinonasal chronic infections, mucoceles, fungal diseases, and granulomatous diseases are then presented, and a recommended imaging protocol is given. PMID- 8419979 TI - Manifestations of AIDS in the oromaxillofacial region. The role of imaging. AB - The primary role of sectional imaging of the oromaxillofacial region in patients who have AIDS is to assess the extent of disease already identified by the clinician. In patients in whom the clinical presentation is complex or confusing, radiographic detection of otherwise occult manifestations of HIV infection plays a key role in the management of the patient with AIDS. The varied manifestations of HIV infection in the oromaxillofacial region and the indications for imaging have been reviewed. PMID- 8419980 TI - Epithelial tumors of the paranasal sinuses and nasal cavity. AB - This article addresses the various epithelial tumors of the nasal cavity and paranasal sinuses. It emphasizes the radiologic evaluation of patients with these tumors and the radiologic findings of importance. The advantages and disadvantages of the use of computed tomographic (CT) and magnetic resonance (MR) imaging in this evaluation are stressed. PMID- 8419981 TI - Nonepithelial tumors of the paranasal sinuses and nasal cavity. Role of CT and MR imaging. AB - This article reviews some of the applications of computed tomographic (CT) and magnetic resonance (MR) imaging in the clinical investigation of nonepithelial tumors and tumorlike lesions of the sinonasal tract. Sixty selected patients primarily with various nonepithelial tumors of the sinonasal tract were included in this study. The MR characteristics of many of these lesions are described. PMID- 8419983 TI - Supplement to Radiology. Volumes 174-185 (I), 1990-1992 and RSNA index to imaging literature. PMID- 8419982 TI - Dental radiology. Role of plain radiographic examination. AB - When a patient presents to a dental office, the imaging modality selected by the dentist is determined by the clinical examination and symptoms. Pain, swelling, or paresthesia usually results in a screening panoramic radiograph. Once a suspicious area is detected, other plain films can be used to provide greater detail. Caries and its sequela and periodontal disease and the surrounding tissues can best be evaluated by intraoral images. In general, today's general dental practice delivers a wide range of preventative and restorative treatments demanding maximum diagnostic information. This can only be accomplished by the dentist having knowledge of the various types of projections that can provide an aid to diagnosis. PMID- 8419984 TI - Role of potassium channels in mitogenesis. PMID- 8419985 TI - Quantitative studies of the structure of proteins in solution by Fourier transform infrared spectroscopy. PMID- 8419986 TI - Hydrophobic characteristics of folded proteins. PMID- 8419987 TI - Chemical threat to birds and mammals in The Netherlands. AB - This article reviews the impact of xenobiotic substances on wild birds and mammals in The Netherlands, as recorded in the field over the past 25 yr. First, a brief survey is given of population trends during this period, using a few illustrative examples to indicate the role played by "environmental policy themes" other than diffusion. The present review of the policy theme diffusion is not intended to be exhaustive, but to provide a broad summary of the nature of the problems involved. Based on this review, conclusions are drawn about the ecological compatibility of various groups of xenobiotics with respect to birds and mammals. Because policy-makers are interested in the relative significance of both the various groups of substances and the various environmental themes for the decline of bird and mammal populations in The Netherlands, a provisional estimate of these contributions is also presented. The main conclusion is that, in the context of the policy theme diffusion, PCBs and pesticides have the greatest impact on birds and mammals. PCBs have an impact up to the species level, whereas the main impact of pesticides today is probably ecological, that is, foraging and habitat changes. It is also concluded that the share of the theme diffusion in the sum total of negative environmental influences is approximately one-quarter. PMID- 8419988 TI - Bioaccumulation and toxicology of chromium: implications for wildlife. AB - The major source of exposure to Cr for wild birds and mammals is through ingestion with food. Chromium(VI) compounds are absorbed significantly more efficiently (2-10% of dose) from the GI tract than inorganic Cr(III) compounds (0.5-3%), due to the increased membrane permeability of the former. Transfer of Cr(VI) into mammalian fetuses has been documented at oral doses of 500 mg Cr/L in drinking water, and injected single doses of 5 mg Cr(VI)/kg BW in dams were teratogenic. Cr concentration data for mammalian and avian wildlife species and their potential food organisms are scarce. Worldwide, fewer than 50 species of free-living mammals and birds have been surveyed with regard to tissue Cr concentrations. Tissue concentrations in animals living in habitats remote from sources of Cr contamination range from approximately 0.1-15 micrograms/g DW depending on the species and tissue analyzed. In habitats experiencing Cr pollution, levels can be up to two orders of magnitude higher. Eisler (1986) suggested that tissue concentrations in wildlife > 4 micrograms/g DW be considered to indicate likely contamination by Cr. Bone tissue often accumulates higher concentrations than other tissues in animals chronically exposed to Cr. Measuring concentrations only in the liver and/or kidneys has been a common practice, yet these organs failed to show evidence of extant Cr contamination in some cases. It is recommended that analysis of the bone, liver, and kidneys be a minimum requirement for future Cr biomonitoring studies. Concentrations in fur or feathers can be extremely variable even among individuals within the same habitat. At best, concentrations in fur and feathers might be used to indicate relative levels of airborne Cr contamination. The toxicological significance of "elevated" Cr concentrations is largely unknown because toxicological data on free-living wildlife species are virtually nonexistent. Based on controlled dosing studies in which Cr compounds were administered orally to experimental animals, dietary Cr concentrations > or = 10 micrograms/g DW in food should be considered potentially harmful to the health and reproductive success of wildlife consumers. Certain species of fish and aquatic invertebrates are sensitive to Cr, showing reduced survival or growth at Cr(VI) concentrations > 10 micrograms/L. The elimination of these organisms from environments contaminated with Cr may have detrimental effects on wild birds and mammals that depend on such organisms for food.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8419989 TI - Decontaminating pesticide protective clothing. AB - The review of recent work on the mechanisms of soil removal from textiles assists in understanding decontamination of pesticide protective clothing. The current work provides explanatory conclusions about residue retention as a basis of making recommendations for the most effective decontamination procedures. A caution about generalizations: Some pesticides produce very idiosyncratic responses to decontamination. An example is the paraquat/salt response. Other pesticides exhibit noticeable and unique responses to a highly alkaline medium (carbaryl), or to bleach (chlorpyrifos), or are quickly volatilized (methyl parathion). Responses such as these do not apply to other pesticides undergoing decontamination. Given this caution, there are soil, substrate, and solvent responses that do maximize residue removal. Residue removal is less complete as the concentration of pesticide increases. The concentration of pesticide in fabric builds with successive exposures, and the more concentrated the pesticide, the more difficult the removal. Use a prewash product and/or presoak. The surfactant and/or solvent in a prewash product is a booster in residue removal. Residues transfer from contaminated clothing to other clothing during the washing cycle. Use a full washer of water for a limited number of garments to increase residue removal. The hotter the washing temperature, the better. Generally, this means a water temperature of at least 49 degrees C, and preferably 60 degrees C. Select the detergent shown to be more effective for the formulation: heavy-duty liquid detergents for emulsifiable concentrate formulations and powdered phosphate detergents for wettable powder formulations. If the fabric has a soil repellent finish, use 1.25 times the amount recommended on the detergent label. For water hardness above 300 ppm, an additional amount of powdered phosphate detergent is needed to obtain the same level of residue removal as obtained with the heavy-duty liquid detergent when laundering fabrics with the soil-repellent finish. The mechanical action of agitation increases dislodgement of particulate material. Too many items in the washing apparatus or too low water volume, or both, decrease agitation and soil removal. Bleach can be used if desired. Fabric softener does not affect pesticide absorption or residue removal in laundering. Dry cleaning is not recommended because the solvents used in dry cleaning may be recycled through dilution, filtration, activated charcoal adsorption, or distillation. Pesticides still may be present in recycled solvents and can be transferred from one item to another, or from one load to subsequent loads of dry cleaning. PMID- 8419990 TI - [What is your roentgen diagnosis? Mycetoma of the foot = maduromycosis, madura foot]. PMID- 8419991 TI - [Research in epidemiology and causality: the example of the Basel Multiple Sclerosis Study 1980 to 1986]. AB - Statistic evaluations on multiple sclerosis 1980 to 1986 in two Swiss cantons have been analyzed, using stochastic mathematical models in order to investigate significance of and possible reasons for variations in incidence. Fluctuation rates of up to 20% could hereby be demonstrated to be due to chance. This could be validated by computer simulations and comparisons with published data. The results show that such apparently large fluctuations in incidence rates should thus be attributed to possible causes with utmost care and prompt for more detailed rules for collection of data. PMID- 8419992 TI - [Is early diagnosis possible in cerebral aneurysm? Symptoms of saccular cerebral aneurysms before the occurrence of massive subarachnoid hemorrhage. Literature review and discussion]. AB - Cerebral aneurysms may manifest as subarachnoid hemorrhage. Approximately two fifth of the patients with subarachnoidal hemorrhage complained of early symptoms prior to the major hemorrhage. The complaints may be headache as the most likely, cranial nerve palsies, different vegetative symptoms, transient neurologic deficits, etc. Considering the good outcome of surgical repair of cerebral aneurysms before a major hemorrhage occurs makes it evident that early diagnosis of intracerebral aneurysm is crucial. In the following we will discuss early signals and warning symptoms with respect to literature and personal experiences. PMID- 8419993 TI - [Cerebral microdialysis in neurological and neurosurgical research and clinical application in neurology and neurosurgery]. AB - Microdialysis of the cerebral extracellular space (CES) provides information on the cortical metabolic state by measurement of the concentrations of metabolites and transmitters as well as of the extracellular pH. In trauma patients, this kind of monitoring should reveal damage from secondary cortical ischemia in an early, hopefully reversible state. The method further allows in vivo measurement of drug levels in the CES and investigation of their effect on metabolism. Although microdialysis has mainly been used experimentally, it is now beginning to be applied clinically. We report on our experience with both experimental and clinical work. PMID- 8419994 TI - Basic research (I) PMID- 8419995 TI - Knuckle-walking ancestors. PMID- 8419996 TI - Conservation: should drug companies share in the costs? PMID- 8419997 TI - Genome project goes commercial. PMID- 8419998 TI - National Institutes of Health. ...Top AIDS official to leave. PMID- 8419999 TI - National Institutes of Health. Gene therapists jump ship... PMID- 8420000 TI - Wake-up call for sleep research. PMID- 8420001 TI - Geneticists trace the DNA trail of the first Americans. PMID- 8420002 TI - Two open complexes and a requirement for Mg2+ to open the lambda PR transcription start site. AB - Potassium permanganate (KMnO4) footprinting in the absence and presence of magnesium (Mg2+) at the lambda PR promoter identified two different open complexes with Escherichia coli E sigma 70 RNA polymerase (designated RPo1 and RPo2). The single-stranded region in RPo1 (formed in the absence of Mg2+) was at most 12 bases long, whereas that in RPo2 (formed in the presence of Mg2+) spanned at least 14 bases. Only in RPo2 did the single-stranded region extend to the start point of transcription (+1, +2). These results provide a structural basis for the requirement for uptake of Mg2+ in the formation of RPo2 from RPo1, as deduced from kinetic studies at this promoter. PMID- 8420003 TI - Molecular mapping and detoxification of the lipid A binding site by synthetic peptides. AB - Endotoxin [lipopolysaccharide (LPS)], the major antigen of the outer membrane of Gram-negative bacteria, consists of a variable-size carbohydrate chain that is covalently linked to N,O-acylated beta-1,6-D-glucosamine disaccharide 1,4' bisphosphate (lipid A). The toxic activity of LPS resides in the lipid A structure. The structural features of synthetic peptides that bind to lipid A with high affinity, detoxify LPS in vitro, and prevent LPS-induced cytokine release and lethality in vivo were defined. The binding thermodynamics were comparable to that of an antigen-antibody reaction. Such synthetic peptides may provide a strategy for prophylaxis and treatment of LPS-mediated diseases. PMID- 8420004 TI - Sequence-specific binding of transfer RNA by glyceraldehyde-3-phosphate dehydrogenase. AB - A transfer RNA (tRNA) binding protein present in HeLa cell nuclear extracts was purified and identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Studies with mutant tRNAs indicated that GAPDH recognizes both sequence and structural features in the RNA. GAPDH discriminated between wild-type tRNA and two tRNA mutants that are defective in nuclear export, which suggests that the protein may participate in RNA export. The cofactor nicotinamide adenine dinucleotide disrupted complex formation between tRNA and GAPDH and thus may share a common binding site with the RNA. Indirect immunofluorescence experiments showed that GAPDH is present in the nucleus as well as in the cytoplasm. PMID- 8420005 TI - Cyclic ADP-ribose in insulin secretion from pancreatic beta cells. AB - Inositol 1,4,5-trisphosphate (IP3) is thought to be a second messenger for intracellular calcium mobilization. However, in a cell-free system of islet microsomes, cyclic adenosine diphosphate-ribose (cADP-ribose), a nicotinamide adenine dinucleotide (NAD+) metabolite, but not IP3, induced calcium release. In digitonin-permeabilized islets, cADP-ribose and calcium, but not IP3, induced insulin secretion. Islet microsomes released calcium when combined with the extract from intact islets that had been incubated with high concentrations of glucose. Sequential additions of cADP-ribose inhibited the calcium release response to extracts from islets treated with high concentrations of glucose. Conversely, repeated additions of the islet extract inhibited the calcium release response to a subsequent addition of cADP-ribose. These results suggest that cADP ribose is a mediator of calcium release from islet microsomes and may be generated in islets by glucose stimulation, serving as a second messenger for calcium mobilization in the endoplasmic reticulum. PMID- 8420006 TI - Blood-brain barrier penetration and in vivo activity of an NGF conjugate. AB - Nerve growth factor (NGF) is essential for the survival of both peripheral ganglion cells and central cholinergic neurons of the basal forebrain. The accelerated loss of central cholinergic neurons during Alzheimer's disease may be a determinant of dementia in these patients and may therefore suggest a therapeutic role for NGF. However, NGF does not significantly penetrate the blood brain barrier, which makes its clinical utility dependent on invasive neurosurgical procedures. When conjugated to an antibody to the transferrin receptor, however, NGF crossed the blood-brain barrier after peripheral injection. This conjugated NGF increased the survival of both cholinergic and noncholinergic neurons of the medial septal nucleus that had been transplanted into the anterior chamber of the rat eye. This approach may prove useful for the treatment of Alzheimer's disease and other neurological disorders that are amenable to treatment by proteins that do not readily cross the blood-brain barrier. PMID- 8420007 TI - Indigenous human cutaneous anthrax in Texas. AB - In December 1988 an indigenous case of cutaneous anthrax was identified in Texas. The patient, a 63-year-old male Hispanic from southwest Texas, was a sheep shearer and had a recent history of dissecting sheep that had died suddenly. He experienced an illness characterized by left arm pain and edema. A necrotic lesion developed on his left forearm, with cellulitis and lymphadenopathy. After treatment with oral and intravenous penicillins, the patient fully recovered. Western blot testing revealed a fourfold or greater rise in antibody titer to Bacillus anthracis protective antigen and lethal factor. This represents the first case of indigenous anthrax in Texas in more than 20 years. PMID- 8420008 TI - Midline disk herniations of the lumbar spine. AB - Most lumbar disk displacements occur through the lateral fibers of the posterior longitudinal ligament, whereas only a small percentage occur through the strong midline fibers. The midline disk herniation is identified as a focal midline dural sac compression and is better seen on the lateral myelogram and computed tomography scan. Of the 22 patients in this study, 14 had midline lumbar disk herniations at L4-5. Their presenting symptoms including back pain with sciatica, were similar to those of lateral disk herniations. On physical examination, however, patients commonly had only positive root tension signs with few nerve root deficits. The incidence of cauda equina syndrome was 27%. The results of treatment were much poorer in these patients than results reported with lateral herniations; 41% were good, 27% fair, and 32% poor. Surgical decompression yielded the best results. A wide exposure is usually needed to provide adequate decompression. PMID- 8420009 TI - Endoscopic transnasal antrochoanal polypectomy: an alternative to the transantral approach. AB - The use of functional endoscopic sinus surgery has been limited typically to management of chronic sinusitis, nasal polyposis, and recurrent acute sinusitis. Antrochoanal polyps (ACPs) traditionally have been resected using a Caldwell-Luc sinusotomy. We used the endoscopic approach, however, in the treatment of five cases of ACP. There was recurrence in one case, but the polyp was successfully removed endoscopically. In the other four cases there has been no evidence of recurrence at a maximum follow-up of 24 months. We believe that transnasal endoscopic antrochoanal polypectomy is an excellent surgical option; there is significantly less postoperative morbidity than with the transantral approach, and rates of complete cure are similar. PMID- 8420010 TI - Carbamazepine therapy in neuropsychiatric patients. AB - We reviewed the charts of 106 patients treated with carbamazepine. Age, sex, race, seizure type, seizure focus, age at onset, duration of epileptic illness, reasons for discontinuing treatment, other medications, and response to treatment of seizures, psychiatric symptoms, and behavioral symptoms were recorded. Statistical analysis was done to determine clinical factors influencing response to carbamazepine. Increasing duration of epileptic illness leads to a decreased response to carbamazepine, which controls both seizure and behavioral symptoms effectively; concurrent medication decreases behavioral symptom control without affecting the control of the seizures. We found a strongly negative correlation between duration of seizures and improvement of psychiatric symptoms with carbamazepine therapy. We provide a clinical profile of the carbamazepine responder to guide clinicians in their use of this drug. PMID- 8420011 TI - Two-way medicine: strategies for improving doctor-patient relationships. AB - Patients are unhappy with the way physicians treat them, not so much medically, as personally. If public opinion is correct, recent medical graduates are less able to win the favor of their patients than previous ones. This article outlines a series of practical strategies for improving doctor-patient relationships. Sooner or later, most physicians learn these strategies by role-reversal. Perhaps a discussion of these with physicians in training (and others) will save some time, generate some goodwill, and improve outcomes for patients and physicians alike. PMID- 8420012 TI - Obstetric anesthesia: delivering quality care. PMID- 8420013 TI - Guidelines for Regional Anesthesia in Obstetrics: potential impact on obstetric anesthesia services in rural Alabama. AB - When the American Society of Anesthesiologists published its Guidelines for Regional Anesthesia in Obstetrics, I did a survey to determine what impact a broadly implemented strict interpretation of those guidelines might have on obstetric anesthesia care in small rural Alabama hospitals. Thirty-six rural Alabama hospitals with fewer than 200 beds were included in the survey, with a response rate of 50%. Data included total deliveries, cesarean section rates, utilization rates of anesthesia services, personnel providing anesthesia care, and identification of physician personnel available during labor and cesarean section. Results showed that anesthesia care is provided for approximately 52% of births in the responding hospitals. Of these, approximately 60% of cesarean sections and 90% of vaginal deliveries are not routinely attended by anesthesiologists. If these data also reflect the nonrespondent hospitals, statewide access could be limited for approximately 6000 parturients annually. The Guidelines should be interpreted with caution. Rather than conform to related policies, interpretation should allow implementation consistent with the capabilities of the individual institutions, while ensuring quality anesthesia care for the parturients. PMID- 8420014 TI - Colorectal cancer: metastatic patterns and prognosis. AB - From 1979 to 1982, 163 patients with colorectal cancer were found to have distant metastases. Of these, 112 (69%) had metastatic disease at the time of initial diagnosis (synchronous metastases [SM]); in the remaining 51 (31%) metastases developed during the course of the disease (metachronous metastases [MM]). The liver was the most common site of metastasis in both groups (72% and 65%, respectively); with the exception of brain metastasis, liver metastasis had the worst prognosis (median survival time [MST], 9 months). The MST for other sites of metastasis were: lung, 10.5 months; bone, 10 months; multiple sites, 10 months; and brain, 5.5 months. Of the 81 patients with SM in the liver, 38% were treated with single modality therapy and 62% with combined modality therapy. Thirty-three patients had MM in the liver. The median time for development of liver metastases (metastasis-free interval [MFI]) was 17.5 months; only lung metastases developed faster (12 months). MFIs for other sites were 20, 20.5, and 33 months for bone, multiple sites, and brain, respectively. PMID- 8420016 TI - Aminoglycoside dosing: a randomized prospective study. AB - Although clinical benefits for aminoglycoside dosing services have been suggested, this has not been clearly documented in a prospective fashion. Therefore, we randomly assigned patients to be dosed (1) by their physician (Physician dosing method), (2) by predicting an initial dosage (Predictive dosing method), or (3) by calculating an initial dosing regimen by measuring the pharmacokinetics for the individual patients after a loading dose (Individual dosing method). The patients' clinical response and nephrotoxicity were then evaluated. The individual dosing method resulted in erratic aminoglycoside levels, necessitating its elimination from the study. This group was not included in the final analysis. Of the 164 patients entering the study, 41 had a documented gram-negative infection, received aminoglycosides for more than 2 days, and had serum aminoglycoside levels measured. The predictive dosing method in these 41 patients produced statistically significant higher peak and lower trough levels, but there was no difference in the incidence of nephrotoxicity or clinical response. The 95% confidence intervals precluded any major clinical benefit in these patients with documented gram-negative infections. We question the previous findings of increased efficacy and decreased nephrotoxicity with the use of an aminoglycoside dosing service and suggest that larger studies be done. PMID- 8420015 TI - Genetic conditions among patients receiving genetic services in middle Tennessee. AB - We reviewed genetics charts of 2235 patients seen from 1985 to 1990 at Vanderbilt University Medical Center, Nashville, Tennessee, and summarized the 20 most common reasons for referral (occurring in 1138 of the patients) and the diagnoses or conditions among patients receiving genetic services in one of four clinical settings (prenatal counseling clinics, general genetics clinics, outreach genetics clinics, and ward consultations). The five most common reasons for referral were advanced maternal age (> or = 35 years) (203/1138, or 18% of patients), followed by dysmorphic features/multiple congenital anomalies (MCA) (185/1138; 16%), developmental delay/mental retardation (MR) (168/1138; 15%), Down's syndrome (103/1138; 9%), and abnormal maternal serum alpha-fetoprotein (MSAFP) (74/1138; 7%). The five most common diagnoses or conditions identified for all genetics patients were advanced maternal age (> or = 35 years) (195/906; 22%), developmental delay/MR (111/906; 12%), dysmorphic features/MCA (107/906; 12%), Down's syndrome (88/906; 10%), and multiple fetal losses (57/906; 6%). Of the 20 most common diagnoses or conditions categorized in 602 of the 906 patients, a multifactorial cause was observed in 25% of those patients; a chromosomal cause was observed in 26% of cases of Down's syndrome, accounting for 55% of the chromosomal disorders; a single gene disorder was observed in 17% of patients; an environmental cause was seen in 4%; and an unknown cause was noted in 28%. We hope this study will help physicians in middle Tennessee and surrounding areas by increasing their awareness of the types and frequencies of genetic diseases so that misdiagnoses and delayed referrals can be avoided. PMID- 8420017 TI - Recent advances in surgery of the temporal bone and skull base. AB - Remarkable advances have been made in the field of neuro-otologic and skull base surgery within the past decade. Each component of the "disease model"- prevention, diagnosis, therapy, and rehabilitation--is undergoing rapid progress. The purpose of this paper is to highlight only a few of these achievements. Each topic chosen has witnessed recent advancement in one aspect of the disease model. Neurofibromatosis (prevention) has been subjected to chromosomal mapping, allowing for genetic counseling. Intraoperative facial nerve monitoring (diagnosis) has allowed improved anatomic and functional preservation of this nerve during surgery. Embolization of glomus tumors (therapy) has rendered these difficult lesions more surgically manageable, and cochlear implantation (rehabilitation) has allowed the profoundly deaf to play a more active role in society. Each of these topics is briefly discussed as it relates to the temporal bone surgeon. PMID- 8420018 TI - Doppler ultrasonography of the umbilical cord in normal pregnancy. AB - Using continuous wave Doppler ultrasonography to measure umbilical artery flow velocity waveforms (FVWs) is a safe, noninvasive means of quantifying placental resistance to blood flow. Detection of abnormally elevated resistance to umbilical perfusion may identify pregnancies complicated by uteroplacental insufficiency. Use of this technique to identify high-risk pregnancies requires ready access to standard tables of normal FVW values obtained from well-defined populations under usual clinical conditions. We report measurements of the systolic-diastolic (S-D) ratio of the umbilical artery obtained between 16 and 43 weeks in 122 normal pregnancies. A gestational-age-adjusted table of normal values is presented. PMID- 8420019 TI - Hand infections. AB - The hand is a commonly injured part of the body that frequently becomes infected. Improperly treated hand infections can lead to severe disability including stiffness, contracture, and amputation. This article describes the current concepts of diagnosis and treatment of common hand infections. PMID- 8420020 TI - Amiodarone pulmonary toxicity: a multidisciplinary review of current status. AB - Amiodarone, a benzofuran derivative, has proven useful in the control of serious cardiac arrhythmias. We reviewed the English language medical literature to characterize clinical, radiographic, scintigraphic, pathologic, diagnostic, and prognostic data concerning amiodarone pulmonary toxicity. Our review showed that features consistent with amiodarone pulmonary toxicity include exertional dyspnea, fever, and high sedimentation rates, usually in patients taking larger maintenance doses. Positive findings on gallium scan, foamy alveolar macrophages on lung biopsy or bronchoalveolar lavage, and resolution of abnormal chest roentgenogram upon withdrawal of amiodarone and/or institution of corticosteroid therapy support a diagnosis of amiodarone pulmonary toxicity. Conversely, maintenance doses of 400 mg or less daily, normal lung diffusing capacity and bronchoalveolar lavage or lung biopsy specimens without foamy alveolar macrophages are features that make amiodarone pulmonary toxicity unlikely. Amiodarone pulmonary toxicity should be considered in any patient who has new or clinical worsening of respiratory symptoms and/or abnormalities on chest roentgenogram. Congestive heart failure is often present in these patients and must be excluded before a diagnosis of amiodarone pulmonary toxicity can be considered. Amiodarone pulmonary toxicity also needs to be distinguished from pulmonary infection. Therefore, amiodarone pulmonary toxicity remains a clinical diagnosis relying upon a composite of clinical, radiographic, and histopathologic findings. PMID- 8420021 TI - Words--10 years later. AB - Certain phrases in the medical language have lost their meaning and in some cases now suggest the exact opposite of the facts. These are "anemia of chronic disease," "liver function tests," "the prevalence," "S1 was present," "S1 is normal," "neurologic syncope," "orthostatic hypotension," "the neck veins were distended," "hospital-acquired pneumonia," and "right-sided backward failure." I suggest a different and more clinically useful way of looking at these phrases. PMID- 8420022 TI - History of pediatrics: Part II. PMID- 8420023 TI - Symptomatic rubella reinfection in an immune contact of a rubella vaccine recipient. PMID- 8420024 TI - Azathioprine in the treatment of chronic refractory steroid-dependent asthma. AB - We have described a patient with chronic, severe, steroid-dependent asthma resistant to all forms of anti-inflammatory therapy. Azathioprine alone effectively improved the patient's symptoms and decreased the daily steroid requirements. Further short- and long-term studies are needed to observe the effects of azathioprine in chronic asthma. PMID- 8420025 TI - What happens when your patient's records are subpoenaed? PMID- 8420026 TI - National Practitioner Data Bank: to report or not? PMID- 8420027 TI - The Nth degree. PMID- 8420028 TI - TMA conference spreads HIV knowledge. PMID- 8420029 TI - CATCHUM Project to develop cancer detection, prevention curriculum. PMID- 8420030 TI - Animal rights group distorts medical facts to further its goals. PMID- 8420031 TI - Clinicopathologic conference: a young man with fever, cough, and chest pain. PMID- 8420033 TI - Hurricane Andrew: a Texas physician helps out. PMID- 8420032 TI - Demographic data of the neonatal intensive care population of a county hospital in Texas. AB - Morbidity and mortality associated with neonatal intensive care affect strongly the socioeconomic aspect of the health-care system. A retrospective study of the neonatal intensive care population at a county hospital in Texas showed that most deaths were related to nontreatable causes. Prematurity was a major cause both of increased mortality and morbidity. Although improved management of a premature infant with hyaline membrane disease using artificial surfactant improved survival, this treatment did not change the morbidity. The incidence of morbidity was related directly to the degree of prematurity. Illicit drug use also had a direct correlation with infants who had low birth weights. Overall, the mortality and morbidity data were consistent with incidence data reported nationally. PMID- 8420034 TI - Endomyocardial biopsies after heart transplantation. The presence of markers indicative of activation. AB - A series of 104 endomyocardial biopsies (EMB) from patients after heart transplantation was evaluated for the presence of immunological markers on graft component and infiltrating cells. This included markers for cells expressing alpha beta-T-cell receptors and gamma delta-T-cell receptors, and cytotoxic T cells with granules bearing the serine esterase Granzyme B; the presence of activation markers identified by CD25 (interleukin 2 receptor), CD30, CD69 (activation inducer molecule), CDw70; macrophages using antibody CD14 (WT14), and cells with Fc gamma-receptors type III (CD16). Almost all cells in T-cell infiltrates expressed the alpha beta-T cell receptor. Cells bearing the gamma delta-T cell receptor were scarcely found. The analysis with respect to the histopathologic diagnosis for rejection showed an absence of significance for T cell subsets, Granzyme B-positive cells, and activation markers except CD25. The numbers of macrophages labeled by CD14 and cells expressing Fc gamma RIII showed a significant relation to histopathology of rejection. Apart from leukocytes, also endothelium in EMB with rejection was labeled by the two anti-Fc gamma RIII antibodies used. In addition, in a small series of biopsies investigated, Fc gamma RI- and Fc gamma RII-positive cells were increased in EMB with rejection, and endothelium was labeled by Fc gamma RII antibodies. A cluster analysis on the basis of scores for CD25, CD14, and anti-Fc gamma RIII revealed three main clusters, one cluster comprising biopsies without abnormalities, one cluster containing EMB with the histopathology of rejection and high scores in immunophenotyping for lymphocytes and macrophages, and one cluster in between. The present data emphasize the importance of macrophage assessment in evaluating pathologic processes during rejection of heart allografts and diagnosing rejection. PMID- 8420035 TI - The effect of different solutions for organ preservation on immediate postischemic pancreatic function in vitro. AB - The present study compares the effect of organ preservation with Euro-Collins solution, cardioplegic histidine-tryptophan-ketoglutarate solution, and University of Wisconsin solution on immediate pancreatic function after cold storage at 4 degrees C for 24 hr. Postischemic organ quality of a porcine pancreas preparation was tested by quantification of physiological and biomedical parameters in a one-line reperfusion system. During reperfusion with a constant arterial pressure the arteriovenous flow rate was significantly higher for HTK (5.7 +/- 0.91 ml/min, n = 8; P < 0.05 vs. EC) and UW (7.4 +/- 0.81 ml/min, n = 8; P < 0.05 vs. EC) than for EC (3.0 +/- 0.26 ml/min, n = 6). The lowest lactate content in the reperfusate was found after HTK protection (HTK, 64.0 +/- 7.2 mumol/50 ml, n = 8; versus EC, 114.2 +/- 1.7 mumol/50 ml, n = 6, P < 0.001; versus UW, 148.0 +/- 28.6 mumol/50 ml, n = 8, P < 0.05). Amylase in the venous effluent was significantly lower (P < 0.05) for HTK or UW protection than for EC (HTK, 189 +/- 72.6 U/ml; UW, 188 +/- 39.4 U/ml; EC, 416 +/- 71.7 U/ml). Oxygen consumption during reperfusion was significantly higher for HTK (2.15 +/- 0.22 microliters/g/min, P < 0.001) and UW (1.80 +/- 0.52 microliters/g/min, P < 0.05) than for EC (0.47 +/- 0.13 microliters/g/min). We conclude that immediate postischemic organ quality and pancreatic function after protection with HTK is not inferior to preservation with UW. PMID- 8420036 TI - Chronic rejection in the rat aortic allograft. V. Mechanism of the angiopeptin (BIM23014C) effect on the generation of allograft arteriosclerosis. AB - Synthetic cyclic octapeptide analogues of somatostatin, such as angiopeptin (BIM23014C; AP) inhibit myointimal proliferation in chronically rejecting rabbit and rat allografts and following angioplasty in rabbits. We have investigated the mechanism of angiopeptin inhibition of allograft arteriosclerosis. DA (RT1a) aortic allografts were transplanted to WF (RT1v) recipients, which either received 80 micrograms/kg/day of AP (Alzet mini-pumps, s.c., 0-180 days) or were left untreated. AP administration did not affect the intensity of adventitial inflammation, nor reduced the disappearance of smooth-muscle cell nuclei from the media (media necrosis); however, it reduced their appearance in the intima and intimal thickening. The effect disappeared, however, from the 3rd month onward. In vivo labeling with tritiated thymidine and autoradiograms demonstrated that AP reduced slightly the proliferation of the inflammatory cells in adventitia and of smooth-muscle cells in the media, and reduced strongly and significantly (P < 0.01) the proliferation of smooth-muscle cells in the intima. Analysis of the major chronic-rejection associated eicosanoids from the vascular wall showed that AP had no effect on the release of the pro-inflammatory thromboxane B2 from the allograft. As AP did not reduce the intensity of perivascular inflammation, reduced only slightly the proliferation of inflammatory cells, and did not affect the release of thromboxane B2 from the inflammatory macrophages, it is likely that the AP effect is not directed to the inflammatory cells. As previous in vitro studies have demonstrated that vascular smooth-muscle cells proliferate in response to several growth factors, and as somatostatin analogues are inhibitory to their action, our data suggest that the action of AP on allograft arteriosclerosis is due to a direct effect on smooth-muscle-cell proliferation. PMID- 8420037 TI - Evidence that anti-CD8 abrogates anti-CD4-mediated clonal anergy but allows allograft survival in mice. AB - Monoclonal antibodies directed against different T cell subpopulations have been used in several rodent models of transplantation to induce long-term unresponsiveness to allografts by a variety of mechanisms. To investigate whether different mechanisms may be operative when different regimens of mAb therapy are used, we studied the effects of various combinations of anti-T-cell antibody treatment on the induction of tolerance in a mouse islet allograft model. Anti CD4 mAb alone, anti-CD8 mAb alone, anti-CD4 mAb plus anti-CD8 mAb, and anti Thy1.2 mAb alone were given at the time of engraftment. Only the anti-CD4 mAb and the anti-CD4 mAb plus anti-CD8 mAb regimens were successful in inducing permanent unresponsiveness to islet allografts. We have previously shown that anti-CD4 mAb alone induces permanent unresponsiveness to islet allografts by a mechanism of clonal anergy, as demonstrated by unresponsiveness of potentially alloreactive T cells to anti-T-cell receptor-specific cross-linking. Interestingly, the potentially alloreactive T cell subsets of recipient mice (V beta 5+ and V beta 11+) made unresponsive to islet allografts by anti-CD4 mAb plus anti-CD8 mAb therapy were not found to be anergic using the same assay. Differences between the repopulation kinetics of CD8+ T cells of anti-CD4 mAb plus anti-CD8 mAb treated recipient mice, which accepted islet allografts, and anti-Thy1.2 treated recipient mice, which rejected islet allografts despite similar levels of initial T cell depletion, suggest that unresponsiveness to alloantigen may have been induced in anti-CD4 mAb plus anti-CD8 mAb treated recipients by clearance of donor passenger leukocytes during prolonged CD8+ T cell depletion. PMID- 8420038 TI - The effect of cyclosporine treatment on the expression of genes encoding granzyme A and perforin in the infiltrate of mouse heart transplants. AB - Following activation of cytotoxic T cells and NK cells several genes encoding proteins putatively involved in cell-mediated cytotoxicity become expressed. The expression of genes encoding the cytotoxic T cell associated serine protease granzyme A and perforin was analyzed in cellular infiltrates of MHC mismatched (H 2d-->H-2k) heterotopic heart transplants both in immunosuppressed recipients treated with cyclosporine and in untreated recipients. Heart transplants were completely rejected by untreated animals on day 10 post-transplantation, whereas CsA treatment generally prolonged survival of the transplants beyond 30 days. In untreated recipients the number of granzyme A- and perforin-expressing cells in heart transplants increased from approximately 10 granzyme A-positive cells/mm2 and 1 perforin-positive cell/mm2 on day 2 posttransplantation to over 80 positive cells for both genes on day 5 posttransplantation. In contrast, these values remained always below 15 positive cells/mm2 for both genes between day 5 and day 30 posttransplantation in CsA-treated recipients. Comparison of the frequency of CD8+ T cells in the infiltrates showed that lower numbers of perforin and granzyme A-positive cells were mainly due to the immunosuppressive action of CsA rather than to reduced infiltration of transplants. The present study shows that expression of granzyme A and perforin gene can be used to discriminate between quiescent and activated cytotoxic cells also in immunosuppressed animals and further confirms that these can be used as sensitive markers for monitoring the fate of a transplant. PMID- 8420039 TI - The use of granzyme A as a marker of heart transplant rejection in cyclosporine or anti-CD4 monoclonal antibody-treated rats. AB - Granzyme A is a serine protease expressed by populations of human and mouse natural killer cells and activated CD4+ and CD8+ cytotoxic lymphocytes; its expression marks a subset of inflammatory cells in allografts, autoimmune diabetes, and a number of other inflammatory lesions. In order to describe more completely the correlation between granzyme A expression and the presence of in vivo cytolytic effects, we grafted allogeneic rat hearts with vascular anastomoses in a heterotopic location, and treated the hosts with either cyclosporine, anti-CD4 monoclonal antibody (MRC OX38), or no therapy. The grafts were evaluated by palpation for cardiac functions, by immunohistochemistry for CD4/CD8 expression, by hematoxylin-and-eosin staining for inflammatory infiltration, and by in situ hybridization for granzyme A expression. The appearance of granzyme A+ cells in untreated allografts preceded both functional and standard histopathological and immunohistochemical evidence of graft rejection by two days. In donor-recipient combinations where cyclosporine and anti-CD4 treatments allowed indefinite allograft survival, the allografts showed minimal numbers of granzyme A+ cells, whether cellular infiltrates developed or not. The number of granzyme A+ cells present in the cardiac allografts in treated and untreated animals correlated with either current or impending episodes of rejection. The early time course of granzyme A expression suggests that it can be used as an early and reliable marker of graft rejection. PMID- 8420040 TI - Transplantation of canine lung allografts preserved in UW solution for 24 hours. AB - To evaluate the efficacy of University of Wisconsin (UW) solution for lung preservation, we performed canine lung allotransplantation with lungs preserved in UW solution. Donor lungs from eight dogs were flushed with UW solution and lungs from four dogs with Euro-Collins solution, and these were preserved for 24 hr in cold storage at 4-6 degrees C. All recipient dogs were given cyclosporine at a dose of 20 mg/kg/day until death. None of the four dogs that received a lung allograft flushed with Euro-Collins solution survived contralateral pulmonary artery and bronchial occlusion. All eight dogs that received a lung allograft flushed with UW solution were alive between 8 and 21 days postoperatively. Six of them survived until pulmonary function tests were performed. The engrafted lungs functioned well until death between days 10 and 21 posttransplantation. The PaO2 was 428 +/- 26 prior to transplantation, 317 +/- 140 at 60 min after transplantation, and 382 +/- 76 mmHg at 10 to 21 days after transplantation. In the other two dogs, the grafted lungs functioned poorly, and macroscopically revealed diffuse organizing pneumonia. We conclude that UW solution is useful for up to 24 hr of cold preservation prior to lung allografting. However, we recommend that lungs preserved with UW solution be transplanted within 24 hr, because, despite adequate function, microscopic damage occurs in the grafted lung. PMID- 8420041 TI - Altered specificity of alloreactive cardiac graft-infiltrating cells by prophylactic treatment with OKT3 or horse antilymphocyte globulin. AB - Graft-infiltrating lymphocytes from patients who were prophylactically treated with OKT3 or horse antilymphocyte globulin (H-ALG) were found to have different specificity patterns from those in the control group that received cyclosporine from the day of transplantation. This prophylactic treatment led to a significant decrease of the HLA-DR-directed cytotoxicity, while the cytolytic response against HLA-class I mismatches was hardly affected. In H-ALG patients without rejection, the percentages of class I-reactive cultures were found to be lower than in the other treatment groups, which was mainly due to a lower percentage of HLA-B--reactive cultures. In CsA and OKT3 patients, cytotoxic T cells were rather directed to HLA-B mismatches than to HLA-A antigens, while in H-ALG patients no difference in HLA-A and B-directed cytotoxicity was found. Our data suggest that OKT3 and H-ALG influence the specificity of the T cell allorepertoire, resulting in a decreased frequency of class II-specific cytotoxic T cells after transplantation. H-ALG also has a downregulating influence on the CTL response against HLA class I (HLA-B) antigens. In some patients a fast regeneration of these cells occurs, which results in a higher rejection incidence during the first posttransplant year. PMID- 8420042 TI - The effect of cryopreservation on the survival and MHC antigen expression of murine islet allografts. AB - Cryopreservation is an effective method of islet storage that can facilitate clinical trials of islet transplantation. In the present study we examined the effect of cryopreservation on the survival of islet allografts and the quantity of islet MHC antigen expression. Islets isolated from CBA/J (H-2k) mice were transplanted into streptozotocin-induced diabetic BALB/c (H-2d) mice treated with or without antilymphocyte serum (ALS). Frozen/thawed (F/T) grafts were cooled slowly to -40 degrees C, stored at -196 degrees C, and thawed rapidly. Fresh and F/T isograft controls reversed diabetes promptly and maintained normoglycemia > 100 days. Allografts of fresh and F/T islets induced normoglycemia initially, but graft failure ensued at 18.2 +/- 1.5 and 16.4 +/- 1.9 days, respectively. ALS treatment prolonged allograft survival significantly to 35.3 +/- 3.9 and 37.5 +/- 6.3 days for fresh and F/T islets, respectively. Following cryopreservation, the quantity of class I antigen expression was reduced by 40%, while the quantity of class II expression was variable. These data indicate that murine islet MHC class I expression is reduced after cryopreservation. This decrease was not associated with altered survival of allogeneic grafts. PMID- 8420043 TI - The effects of cyclosporine on HILDA/LIF gene expression in human T cells. AB - We examined the effect of cyclosporine on HILDA/LIF gene expression in alloreactive human T lymphocyte clones (ATLCs) 2B11 and 2F7 obtained from cells infiltrating a rejected human kidney graft. Both ATLCs were stimulated either by the specific antigen or by PMA + calcium ionophore in the presence of various concentrations of CsA (10-500 ng/ml). Inhibition of HILDA/LIF gene expression was analyzed at the protein level using a proliferative assay on the HILDA/LIF dependent Da-1a cell line and by RNA blotting using a specific probe. Without CsA, the kinetics of mRNA accumulation for both ATLCs peaked at 5 and 10 hr, respectively, after mitogenic and antigenic stimulations. HILDA/LIF activity peaked at 24 and 72 hr, respectively, after mitogenic and antigenic stimulation in supernatants from both ATLCs and decreased thereafter. Subsequent experiments with CsA were thus performed at these time points. Our results show that HILDA/LIF mRNA accumulation and protein secretion in 2B11 and 2F7 clones were strongly inhibited in a dose-dependent manner by CsA, in both stimulation conditions. Maximal inhibition of HILDA/LIF transcripts and protein secretion (60 90%) was observed within the range of 75-500 ng/ml CsA. PMID- 8420044 TI - Evidence that polyreactive antibodies are deposited in rejected discordant xenografts. AB - The rejection of organs transplanted between phylogenetically disparate species is thought to be initiated by the binding of naturally occurring antibodies of the recipient to the endothelium lining the blood vessels of the donor organ. We recently showed that among the xenoreactive natural antibodies in human serum that react with porcine endothelial cells and endothelial cell--derived glycoproteins are polyreactive antibodies. To test whether polyreactive natural antibodies are present in rejected xenografts we developed a series of hybridoma derived antibodies specific for the polyreactive human monoclonal antibody 103, which we have shown to bind efficiently to porcine endothelial cells. Using these antiidiotypic reagents we detected antibodies bearing the 103 idiotype in a panel of human sera and on the surface of lymphocytes in the spleen of humans, rhesus monkeys, and baboons. The antiidiotypic reagents reacted in a pattern similar to the distribution of IgM, with immunoglobulin deposits in tissues obtained from pig-to-rhesus monkey and pig-to-baboon xenografts. Analysis of immunoglobulin eluted from these sites showed that they antibodies display the antigen-binding features of natural antibodies. Our findings are consistent with the hypothesis that idiotypes of polyreactive natural antibodies are broadly crossreactive. They also suggest that the polyreactive and xenoreactive IgM, which have been detected based on in vitro assays, may be deposited in a xenogeneic organ graft. PMID- 8420045 TI - Tolerance by old livers of prolonged periods of preservation in the rat. AB - To determine the tolerance of old livers to prolonged preservation, livers from aged rats (corresponding to humans in the sixth to seventh decades of life) were transplanted after specific periods of cold preservation. Male BN/BiRij rats received orthotopic, arterialized liver grafts from either young (5 months) or old (25 to 28 month) donor rats after liver storage for 12 (n = 6), 24 (n = 6) or 30 (n = 10) hours in University of Wisconsin solution. Outcome was assessed by survival, liver enzymes after transplantation, and histology of the grafts. There were no significant differences in survival rates between recipients of old and young grafts. All rats survived after 12-hr and 24-hr preservation except one recipient of an old graft preserved for 24 hr. After 30-hr preservation recipients of old and young livers had identical survival rates (60%). There was a strong correlation between the highest postoperative AST and ALT and the duration of preservation in all groups (P < 0.0001), but only in the 24-hr preservation experiments was the ALT significantly higher in recipients of old grafts than in recipients of young livers (P = 0.025). Age of the donor did not significantly affect the peak AST, but there was a correlation between donor age and the highest postoperative ALT (P = 0.007). Although intracellular vacuolization was a prominent histological finding in more than half of the old livers at the end of preservation, it was not associated with an increase in mortality. It is concluded that under the ideal conditions provided in the experiments, old rat livers tolerate long preservation periods with satisfactory graft survival compared with young livers. PMID- 8420046 TI - Evidence inconsistent with a role for tumor necrosis factor in tumor allograft rejection. AB - Since it has been suggested that tumor necrosis factor is involved in organ allograft rejection, experiments were undertaken to determine whether it is involved in the rejection of allogeneic P815 tumor cells growing as ascites in the peritoneal cavities of mice. The results show that biologically active TNF was not detectable in serum or ascites fluid of mice during either growth or vigorous rejection of the tumor. On the other hand, mice in the process of rejecting their ascites tumor displayed a greatly enhanced capacity to produce TNF in the peritoneal cavities and systemically in response to an injection of endotoxin i.p. These results show that the immune response to the tumor was associated with the priming of host cells for increased TNF production both locally and systemically in response to an appropriate stimulus that was not supplied during the generation and expression of immunity. Additional evidence against a role for TNF in ascites tumor allograft rejection is seen in the finding that i.p. administration of anti-TNF antibodies failed to interfere with elimination of allogeneic tumor cells, even though the antibodies were given repeatedly before and during the rejection process. Taken together, these results are inconsistent with the view that TNF is involved in the effector stage of the anti-allograft response, even though cells at the site of rejection are primed to produce large quantities of TNF in response to endotoxin. PMID- 8420047 TI - Enzyme-linked immunoassay for anti-HLA antibodies--an alternative to panel studies by lymphocytotoxicity. AB - In order to provide a simple and specific assay for the detection and quantitation of IgG and IgM anti-HLA antibodies in sera, HLA antigens purified from a pool of 240 random donor platelets were used to develop a solid-phase enzyme-linked immunoassay (EIA). The reference values for identifying the presence of IgG or IgM anti-HLA antibodies were determined by assaying sera from 39 healthy individuals without prior HLA alloimmunization. The assay was evaluated by studying sera from 122 patients who had been characterized previously for panel reactive antibodies by the lymphocytotoxicity assay (LCA). A significant linear correlation between two assays was noted (r = 0.8, P = 0.0001). Further analyses of the data demonstrated that the newly developed EIA has 100% specificity and 95.3% sensitivity as compared with the LCA. Additional studies revealed that patients whose PRA increased or decreased over time were in parallel with antibody levels measured by EIA. When the EIA was used to measure anti-HLA antibody titers, it was more sensitive than the LCA. Since the EIA is sensitive, specific, and technically less demanding, it should provide an useful alternative to reduce the number of the more laborious panel studies for monitoring anti-HLA antibody status in candidates for organ transplantation and recipients of blood transfusions. PMID- 8420048 TI - Specific prolongation of skin graft survival following retroviral transduction of bone marrow with an allogeneic major histocompatibility complex gene. AB - Engrafted allogeneic hematopoietic cells have a unique capacity to induce a state of donor-specific transplantation tolerance across major histocompatibility complex barriers. This state allows permanent acceptance of donor-type organ grafts, with otherwise normal immunocompetence. We hypothesized that introduction of allogeneic MHC genes into autologous bone marrow which is then returned to recipient mice might similarly induce specific tolerance to products of the introduced MHC genes, without the risk of graft-vs-host disease. We demonstrate here that the introduction of MHC class I Kb cDNA by retrovirus-mediated gene transfer into B10.AKM (Kk) hematopoietic cells confers specific hyporesponsiveness to allogeneic skin grafts expressing Kb. PMID- 8420049 TI - A comparison of different methods of estimating glomerular filtration rate in cyclosporine-treated renal transplant patients. PMID- 8420050 TI - Obliterative bronchiolitis following single-lung transplantation--diagnosis by spirometry and transbronchial biopsy. PMID- 8420051 TI - A randomized trial of deflazacort versus 6-methylprednisolone in renal transplantation--immunosuppressive activity and side effects. PMID- 8420052 TI - Transient deterioration of intrapulmonary shunting after pediatric liver transplantation. PMID- 8420053 TI - Membranous allograft nephropathy. Remission of nephrotic syndrome with pulsed methylprednisolone and high-dose alternate-day steroids. PMID- 8420054 TI - Accelerated liver allograft rejection during prophylactic immunosuppression with OKT3. PMID- 8420055 TI - Prevention of graft-versus-host disease by RS-61443 in two different rodent models. PMID- 8420056 TI - Restoration of pancreas graft function preserved by a two-layer (University of Wisconsin solution/perfluorochemical) cold storage method after significant warm ischemia. PMID- 8420057 TI - 96-hour cold-storage preservation of the canine pancreas with oxygenation using perfluorochemical. PMID- 8420059 TI - Organ procurement. PMID- 8420058 TI - The influence of the temperature of storage on survival and ultrastructure of transplanted UW-preserved rat liver grafts. PMID- 8420060 TI - Organ procurement. PMID- 8420061 TI - Attenuation of warm ischemic injury of rat lung by inflation with room air- assessment of cellular components and the surfactant in the bronchoalveolar lavage fluid in relation to changes in cellular adenosine triphosphate. AB - Studies were made on the effects in rat lungs of aerobic and anaerobic conditions on the intracellular levels of adenosine triphosphate and its related metabolites, the releases of intracellular enzymes, and the secretion of pulmonary surfactant. After warm ischemia for 120 min, the ATP content of lungs inflated with air was significantly higher (8.0 +/- 1.2 mumol/g dry weight) than those of deflated lungs and lungs inflated with nitrogen (0.8 +/- 0.7 mumol/g dry weight and 2.0 +/- 0.7 mumol/g dry weight, respectively; P < 0.001). The amounts of intracellular enzymes, such as lactate dehydrogenase, cytosolic and mitochondrial aspartate aminotransferase, and protein in the bronchoalveolar lavage fluid (BALF) of air-inflated lungs were significantly less than those in BALFs of deflated and nitrogen-inflated lungs (P < 0.001). The BALF-contents of dipalmitoyl phosphatidylcholine (DPPC), the main component of alveolar surfactant of aerobic and anaerobic ischemic lung were, however, similar. During 120-min warm ischemia after lavage, air-inflated lungs secreted significantly more DPPC into the alveolar space than nitrogen-inflated lungs did (P < 0.001). We conclude that cell membranes in the lungs are damaged under anaerobic conditions, but that inflation of ischemic lungs with air is effective for protecting them from cell injury and for maintaining the intracellular level of ATP and the ability of the cells to secrete pulmonary surfactant. PMID- 8420062 TI - Urocanic acid as an immunosuppressant in allotransplantation in mice. AB - Urocanic acid (UCA) was investigated for its activity as an immunosuppressive agent in murine heterotopic allotransplantation. Mice grafted with cornea, pancreas, and skin under the renal capsule were given either intravenous, subcutaneous, or topical cis-UCA in the peritransplant period. Grafts were performed across complete and minor MHC barriers. Peritransplant immunosuppression with cis-UCA prolonged survival of these grafts, but no route of administration was clearly superior to another. Survival of cornea was prolonged more than survival of skin or pancreas in all MHC disparate combinations. cis-UCA was also used to assess its effect on stimulator cells, derived from mice that had received UCA, in mixed lymphocyte responses. cis-UCA adversely affected the ability of stimulator cells to provoke T cell proliferation in an allogeneic MLR. These findings suggest that cis-UCA may be a potentially useful immunosuppressive agent. PMID- 8420063 TI - A randomized clinical trial of induction therapy with OKT3 in kidney transplantation. AB - A randomized, prospective multicenter trial was conducted to compare the safety and efficacy of OKT3 as an induction therapy with that of conventional immunosuppressive therapy administered to cadaveric renal allograft recipients. Two hundred fifteen patients were treated either with OKT3 plus azathioprine and steroids for 14 days with the delayed addition of cyclosporine on day 11, or with conventional immunosuppression (steroids, azathioprine, and cyclosporine). OKT3 patients had significantly fewer rejection episodes (51% vs. 66%, P = 0.032), a longer time to initial rejection (46 days vs. 8 days, P = 0.001), and fewer rejection episodes per patient (0.82 vs. 1.14, P = 0.014) than conventionally treated patients. Kaplan-Meier estimates of two-year graft and patient survival rates were 84% and 95%, respectively, for the OKT3-treated group, and 75% and 94%, respectively, for the conventionally treated group. Following a subsequent first rejection episode, OKT3 reversed 93% of the rejections in patients receiving OKT3 induction therapy and 94% in patients receiving conventional therapy. Adverse experiences reported during OKT3 induction therapy were similar to those seen when the drug was used for rejection. Following initial exposure, 40.3% of the patients tested were positive for anti-OKT3 antibody, only 6.7% of which were of high titer (1:1000). In the presence of low titer (1:100 or less) antibody, OKT3 was successful in reversing rejection in five of six retreated patients tested. In conclusion, treatment with OKT3 (in combination with azathioprine, steroids, and the delayed addition of cyclosporine) is an effective approach for renal allograft maintenance. PMID- 8420064 TI - A study of treatment compliance following kidney transplantation. AB - Kidney transplantation is a successful treatment for end-stage renal disease. We studied demographic and psychosocial variables that relate to compliance behaviors following renal transplant. One hundred and five renal allograft recipients, with a minimum of 18 months follow-up, were studied. A biographical questionnaire, the Center for Epidemiologic Studies Depression Scale, the Multidimensional Health Locus of Control Scale, and the Social Support Appraisals Questionnaire were used as measuring instruments. Specifically for this study, we designed a Health Belief Model Questionnaire, a Patient and Provider Relationship Questionnaire, a Compliance Self-Report Questionnaire, and a Self-Efficacy Questionnaire. Compliance was determined by cyclosporine whole blood levels > 30 ng/ml, maintenance of ideal body weight (< 20% gain), and percentage of missed clinic visits (< 20%). Data was analyzed using discriminant analysis, Pearson's correlation, and chi-square. Four groups were identified, i.e., overall compliant (n = 25), noncompliant with diet (n = 29), noncompliant with medication (n = 27), and overall noncompliant (n = 29). No patient missed > 20% of clinic visits. Discriminant function analysis distinguished patients who were compliant from those who were not. Males were more likely to be noncompliant with medication, whereas females were more likely to be noncompliant with diet. Noncompliance was also associated with increased numbers of prescribed medications, depression, black race, locus of control attributed to powerful others, unemployment, as well as the perceived amount of social and family support. Patients with failed grafts (n = 14) were more depressed (P < 0.05), perceived less benefit from the treatment regimen (P < 0.01), and had less confidence in their care providers (P < 0.05) than those recipients of successful grafts (n = 91). In conclusion, this study identifies a number of psychosocial and demographic variables that impact on patient compliance behaviors after renal transplant. Interventional strategies to obviate noncompliance will need to consider these heterogeneous variables in order to maximize long-term renal allograft survival. PMID- 8420065 TI - A randomized double-blind trial of the use of human recombinant superoxide dismutase in renal transplantation. AB - Oxygen free radical generation has been implicated as a possible mediator of the reperfusion injury postulated to occur following revascularization of the cold preserved and transplanted kidney. The superoxide radical (O2-) scavenger, superoxide dismutase, from bovine or recombinant (rh-SOD) sources, may ameliorate oxygen-free-radical mediated reperfusion injury of transplanted kidneys. To test this hypothesis, we performed a prospective, randomized, double-blind trial of the use of human rh-SOD in renal transplantation at three participating centers. Half of a 20 mg/kg solution of rh-SOD or placebo was administered as a bolus intravenous injection immediately prior to renal allograft reperfusion and the remainder as a peripheral intravenous infusion for 1 hr thereafter. Posttransplant renal function was determined using 99Tc-DTPA clearance to measure glomerular filtration rate at 48 +/- 24 hr and day 6 post-transplant. A two tailed t test was used for pooled data, and analysis of variance was used to evaluate between center differences in outcome. One hundred and sixteen patients (58 rh-SOD and 58 placebo) were entered into the study. No adverse reactions to rh-SOD or placebo were noted. No differences were noted between rh-SOD and placebo groups with regard to GFR at 48 hr, serum creatinine or creatinine clearance at day 6, or percentage of patients with GFR < or = 10 ml/min or < or = 5 ml/min at 48 hr. The data did not vary when analyzed by center or in aggregate form, and no correlation was noted between storage time and GFR in either group. We conclude that data from this trial provide little basis for the use of rh-SOD as described to ameliorate reperfusion injury in transplanted kidneys. PMID- 8420066 TI - Interleukin 2 immediately after autologous bone marrow transplantation for acute lymphoblastic leukemia--a phase I study. AB - Despite intensive conditioning and marrow purging, leukemia relapse frequently follows autologous BMT for acute lymphoblastic leukemia. To generate antileukemic immunologic activity, we performed a phase I study using recombinant human interleukin 2 given immediately posttransplantation. This early period was chosen because of low disease burden; therefore induced in vivo effector:target cell ratios might be most favorable. IL-2 was given by continuous infusion (96 hr/week x 3 weeks) beginning day +1. Fourteen patients with high-risk ALL were treated at 0.5, 1.0, and 2.0 x 10(6) U/m2/day IL-2. The clinical toxicity, hemopoietic recovery, and immune activation in the IL-2-treated patients was compared with that in a group of autologous BMT patients receiving no IL-2. The patients receiving IL-2 had a trend toward earlier neutrophil, platelet, and RBC recovery plus earlier hospital discharge versus non-IL-2 controls. However, IL-2 plus the inherent toxicity of transplantation often produced hepatic, pulmonary, and renal toxicity. Assessment of immune activation induced by in vivo IL-2 (following 3 weeks of IL-2) showed proliferation of CD8+ T cells having in vitro cytotoxicity against the Nalm-6 ALL cell line in most patients. Little enhancement of natural killer activity by immunophenotype or cytotoxicity against K562 cells was observed. IL-2 given immediately post-BMT induces infrequent but significant toxicity at lower doses than in the non-BMT setting. This toxicity may result from pre-BMT conditioning in conjunction with T cell activation. The immunotherapeutic potential, dose, and schedule of IL-2 following BMT require further study along with measures to reduce its toxicity. PMID- 8420067 TI - Acute renal failure and degenerative tubular lesions associated with in situ formation of adenovirus immune complexes in a patient with allogeneic bone marrow transplantation. AB - We describe the development of acute renal failure and degenerative tubular lesions associated with local immune deposits in a patient with allogeneic bone marrow transplantation. A 21-year-old man with an acute myelocytic leukemia received a bone marrow graft from a cousin mismatched for a single HLA-DR locus antigen. Hemorrhagic cystitis due to adenovirus type 11 infection occurred 26 days after transplantation, and 17 days later the patients developed acute renal failure. A study of renal tissue obtained by needle biopsy showed degenerative and necrotic lesions, especially in the distal part of the nephron. By electron microscopy adenovirus type 11 particles were found in the nuclei of tubular cells and in cellular debris in tubular lumina. By immunofluorescence technique, granular immune deposits containing adenovirus type 11 related antigen(s), immunoglobulins, C3, and membrane attack complex (MAC) C5b-9 of the complement system were detected along the tubular basement membranes but not in glomeruli. The patient's IgG did not bind to normal human kidneys. These findings suggest that adenovirus type 11 directly induced acute tubular damage, and that the tubular immune deposits were formed "in situ" by viral antigens and circulating viral antibody. PMID- 8420069 TI - The synergistic effects of cyclosporine and endothelin--demonstration of an important cardiodepressor action. AB - Cyclosporine represents the foundation for current immunosuppressive therapy following solid organ transplantation. CsA use is associated with renal insufficiency and systemic hypertension. We hypothesized that CsA would enhance the vascular actions of endothelin (ET). Three groups of anesthesized dogs (n = 15) were studied. Group 1 received CsA alone (1 mg/kg), group 2 received ET alone (1 ng/kg/min), and group 3 received combined CsA (1 mg/kg) and ET (1 ng/kg/min). The hemodynamic and renal effects were evaluated after 30 min. Combined treatment resulted in a profound reduction in mean arterial pressure (-62 +/- 14 mmHg (P < .05) and cardiac output (-2.2 +/- 0.4.1/min (P < .05). The reduction in mean arterial pressure and cardiac output were significantly greater than that observed with CsA or ET alone. Systemic vascular resistance was not significantly changed. Combined CsA and ET resulted in a significant reduction in renal blood flow (195 +/- 18 to 101 +/- 11 ml/mm P < .05) but without evidence of active renal vasoconstriction. The decline in GFR (31.8 +/- 5.6 ml/min to being unmeasurable) was of greater magnitude than the change in renal blood flow, suggesting enhanced afferent anteriolar vasoconstriction or an alteration in the ultrafiltration coefficient. These studies demonstrate an important and synergistic cardiodepressor effect when CsA and ET are combined. PMID- 8420068 TI - In vitro alloreactivity against host antigens in an adult HLA-mismatched bone marrow transplant recipient despite in vivo host tolerance. AB - An adult recipient of an HLA-DR, DQ-mismatched, T cell-depleted bone marrow graft, who remains without graft versus host disease and who is not maintained on immunosuppressive therapy, was studied at 23 months posttransplantation for in vitro reactivity against the mismatched antigens of the host. The donor's PBMC's proliferated vigorously against the recipient's stimulators in the pretransplant mixed lymphocyte cultures (MLC). After transplant reconstitution, MLCs demonstrated that the in vitro response of engrafted donor T cells against host MHC class II antigens was equivalent to control allogeneic responses, while there was no detectable response against the donor's antigens. Posttransplantation limiting dilution analysis showed no difference between the precursor frequencies of antihost responders among populations of fresh donor PBMCs and among the engrafted cells of donor origin that are found circulating in the patient. This result suggests that clonal deletion is, at best, incomplete and that peripheral tolerance is essential in protecting this patient from GVHD. These findings also support the conclusion that bone marrow-derived thymic elements may be important for clonal deletion in human chimeras. PMID- 8420070 TI - OKT3 rescue for steroid-resistant rejection in adult liver transplantation. AB - The results of OKT3 use for steroid-resistant rejection rescue in adult liver transplantation were analyzed retrospectively from a single transplant center. Comparison was made with concurrent patients who had no rejection (NR) or steroid responsive rejections (SR). The records of 290 patients who underwent 323 liver transplants from April 1985 to December 1989 were examined. The first technically successful grafts were used for this analysis (265 grafts). Follow-up was a minimum of 1 year, or until death or loss of graft. All patients received triple drug induction immunosuppression (CsA, Aza, steroids). Initial rejection was treated with 1 g methylprednisolone bolus i.v., followed by a 5-day taper of steroids from 200 mg to 20 mg. No rejection occurred in 108 (40.8%) and SR in 86 (32.4%), and OKT3 was given for persistent rejection in 71 (26.8%). The age, sex distribution, mean follow-up, and preoperative status were similar in all three groups. The preoperative diagnoses were similar, except for fulminant liver failure, in which 19 of 20 patients experienced rejection (P < 0.0001). The median hospitalization stay was 37 days for OKT3, 27 days for SR, and 21 days for NR (P < 0.0001). The median ICU stay was similar in the three groups (OKT3, 4; SR, 4; NR, 3). Infections in the first 6 weeks, and in the period of 6 weeks to 1 year posttransplant, were of similar frequency for all three groups. By the Kaplan-Meier estimation, the graft and patient actuarial survival rates were comparable. At 1 year, the graft survival rate was 79.6% for NR, 79.8% for SR, and 67.6% for OKT3. The 1-year patient survival rate was 85.2% for NR, 83.7% for SR, and 84.5% for OKT3. Following treatment by OKT3, rejection was permanently reversed in 42 patients. A temporary response occurred in 12 patients, 16 patients failed to respond to OKT3, 2 patients died during therapy, and 6 of the nonresponders died within 12 months. Additional OKT3 treatment was attempted in 6 patients for persistent rejection within a 1-month interval from the previous OKT3 course. Of these 6, 4 developed lymphoproliferative disorder, and only 1 survived in response to drastic reduction of immunosuppression. In conclusion, OKT3 was effective as rescue therapy for adult liver transplant steroid-resistant rejection. Because of the associated morbidity and expense, OKT3 should be used in a selective fashion. Failure to respond to OKT3 is a serious complication, and should not be managed by prolonged or repeated courses, but rather by alternative means. PMID- 8420071 TI - Arterial ketone body ratio as a predictor of donor liver viability in human liver transplantation. AB - The viability of the donor liver was assessed with regard to early postoperative survival in human liver transplantations from 40 brain-dead donors at Hannover Medical College and 13 living donors at Kyoto University by measuring the arterial ketone body ratio (AKBR). Of 40 grafts harvested from brain-dead patients in Hannover, 35 survived the first week after operation, but 5 developed initial nonfunction of the transplanted graft within the first week. The mean AKBR values were 1.11 +/- 0.11 for grafts that survived and 0.44 +/- 0.10 for grafts that failed (P < 0.01). The AKBR values of the 5 initially nonfunctioning cases were all below 0.7. Of 13 grafts harvested from the living donors in Kyoto, all survived the first week. The AKBR values of the donors were all above 1.0, with a mean value of 1.87 +/- 0.23. Among all 53 cases, the survival rate of the grafts with AKBR above 0.7 was significantly higher than that of the grafts with AKBR below 0.7 (100% vs. 62%, P < 0.01). No other donor parameters, including age, dose of dopamine administered, and clinical laboratory findings, were significantly related to differences in graft survival rates. AKBR is a useful index for the evaluation of donor liver viability. Grafts used from donors with AKBR of less than 0.7 have a significantly increased risk of early nonfunction. Grafts from donors with AKBR of greater than 1.0 have, in our experience, always been viable after transplantation. PMID- 8420072 TI - The effect of cyclosporine on the progression of human immunodeficiency virus type 1 infection transmitted by transplantation--data on four cases and review of the literature. AB - Two women and two men were infected with the human immunodeficiency virus type 1 (HIV-1) transmitted by renal transplantation from i.v. drug-addicted donors in 1984. The four recipients were treated with cyclosporine and methylprednisolone (one patient only for three months because of early graft failure). Two patients died 66 and 74 months after transplantation, one of endocarditis and one of cerebral hemorrhage. Despite several infections including urinary tract infection (n = 8), peritonitis (n = 1), shunt infection (n = 1), bronchitis (n = 1), salmonellosis (n = 1), herpes stomatitis (n = 2), herpes zoster (n = 1), and cytomegalovirus (n = 1), and despite treatment of several rejection episodes (n = 8), none of them had or has infections typical of the acquired immunodeficiency syndrome (AIDS). However, two patients developed cervical lymphadenopathy and one autoimmune thrombocytopenia 15-20 months after HIV-1 infection. Their T helper cell counts (355/microliters to 75/microliters) and helper/suppressor T cell ratios (1.0-0.2) are distinctly lowered. One patient has membranous glomerulopathy with virus-like particles within and on the outside of the basement membrane and tubuloreticular inclusions in glomerular endothelial cells. We evaluated the case reports of 53 patients with HIV-infection caused by an infected transplant or by blood transfusions during or shortly after transplantation. The cumulative incidence of AIDS was significantly lower in 40 transplant patients with an immunosuppressive regimen including cyclosporine than in 13 transplant patients receiving immunosuppressive treatment without cyclosporine (5-year cumulative risk of AIDS: 31% versus 90%, P = 0.001). PMID- 8420073 TI - Treatment of organic impotence with penile prosthesis in renal transplant patients. AB - Seven patients were identified who underwent both renal transplantation and penile prosthesis implantation at our institution between June 1980 and June 1990, and their charts were retrospectively reviewed. A total of nine penile prostheses were placed in these patients, five prior to transplantation and four following transplantation. One patient received two prostheses prior to transplantation. One patient received a prosthesis both before and after transplantation. No complications were seen in the four prostheses placed following transplantation with a follow-up of one to forty months (mean 18 months). Of the five prostheses placed prior to transplantation, two were removed due to periprosthetic infections, a cylinder leak developed in one, and one was complicated by penile and scrotal erythema with sepsis. PMID- 8420074 TI - Spontaneous rupture of human renal allografts. AB - Preservation of renal allografts following spontaneous rupture is a surgical challenge. Salvage was attempted in 3 patients. Despite successful repair in all 3, transplant nephrectomy was ultimately required in 2 patients because of irreversible rejection. The third patient is doing well forty-eight weeks after transplantation. PMID- 8420075 TI - Results of window operation for primary hydrocele. AB - The window operation for hydrocele repair has been reported to be a safe out patient procedure, with few complications and relapses. In this pilot study the procedure could be safely done on an out-patient basis and the rate of complications (8%) was comparable to that of other procedures. However, 21/23 evaluated hydroceles relapsed and retreatment with a second window operation led to further relapses within a short time. Eight patients with 11 hydroceles required another procedure. We have therefore abandoned the window operation for hydrocele repair. PMID- 8420076 TI - New matrix material for potential use in "reversible" vasectomy. Preliminary animal biocompatibility studies. AB - The biocompatibility of a new elastomeric-hydrogel matrix biomaterial for use as an intravasal occlusive device was assessed by inserting it into the vas deferens of dogs. The vas was removed and examined histologically after varying periods of time. The biomaterial resulted in total occlusion of the vas. Epithelial changes were limited to squamous metaplasia in areas adjacent to the implant. Changes in the subepithelium were minimal. This new material is biocompatible with the dog vas, and it has potential as an occlusive device for "reversible" vasectomy in men. Semen analysis studies are needed to assess the completeness and reversibility of the vasal occlusion achieved with this implant. PMID- 8420077 TI - Endoscopic incision of persistent ureteral infoldings. AB - We report 2 cases of persistent ureteral infolding in a four-month-old infant and an eight-year-old boy, both presenting with hydronephrosis. Initial diagnostic evaluation showed multiple pleats in the upper ureter. Endoscopic incision of the pleats relieved hydronephrosis. The concept of persistent ureteral infolding seems to apply to these cases. PMID- 8420078 TI - Transvesical anastomosis after radical retropubic prostatectomy. PMID- 8420079 TI - The 6.9 F semirigid ureteroscope in clinical use. AB - The 6.9 F semirigid ureteroscope has a hybrid design that shares the rigid ureteroscope body with the fiberoptic imaging bundle of the flexible ureteroscopes. This allows more space for the inner channel, so that there are two working channels of 3.4 F and 2.3 F. This ureteroscope was used in 65 cases on 57 patients without dilatation of the intramural ureter. It was mainly used for the management of ureteral calculi (29 cases) and for visual diagnostic purposes (28 cases). The double-channel design proved extremely helpful in lithotripsy, while the ability to access the ureter without dilatation gave a consistently clearer picture for diagnostic purposes. Forty-two cases were done under local anesthesia with sedation. In 1 patient a minor perforation developed secondary to use of electrohydraulic lithotripsy (EHL). Postoperatively, 7 patients complained of colic, and in 5 patients a low grade fever developed, while 2 patients had both. PMID- 8420080 TI - Unusual case of renal Candida fungus balls. AB - Renal fungus balls are rare in the adult population. A case of asymptomatic renal pelvic fungus balls is presented, and several approaches to treatment are discussed. PMID- 8420081 TI - Chickenpox in adult presenting as acute severe genital infection. PMID- 8420082 TI - Extracorporeal shock-wave lithotripsy in transplanted kidney. AB - Urolithiasis is the least described urologic sequela of renal transplantation. We describe a renal transplant patient who presented with painless gross hematuria. An intravenous pyelogram demonstrated a 4 x 7-mm calculi in the region of the ureteropelvic junction, causing moderate hydronephrosis. The patient was treated successfully with extracorporeal shock-wave lithotripsy (ESWL). Serum creatinine and twenty-four-hour creatinine clearance were unchanged from levels prior to ESWL. PMID- 8420083 TI - Ischemia of neobladder. Clinical manifestations and management. AB - Substitution enterocystoplasty is becoming more common in a selected group of patients. Acute or chronic ischemia is an exceptional event. We describe the clinical manifestations and management of chronic ischemia of a neobladder that occurred weeks after a successful initial operation. The possible mechanisms responsible for this complication are reviewed. PMID- 8420084 TI - Renal angiomyolipoma associated with neurofibromatosis and primary carcinoid of mesentery. AB - A patient with von Recklinghausen's disease manifested by dermal neurofibromatosis and cafe-au-lait spots presented with complaints of malaise, weight loss, lower extremity weakness, and a palpable left lower abdominal quadrant mass. Evaluation revealed a lumbar neurofibroma, a localized primary carcinoid tumor of the mesentery, and a left renal angiomyolipoma. Although an association between neurofibromatosis and carcinoid has been previously reported, we believe this is the first report documenting the association of all three entities. PMID- 8420085 TI - Primary malignant melanoma of urinary bladder. AB - We report a case of primary malignant melanoma of the urinary bladder. Exclusion of other sites of melanoma confirmed the primary character of the tumor, and careful histologic examination proved it to be malignant melanoma. Six previously reported cases of bladder melanoma are reviewed. Therapy and prognosis are discussed. PMID- 8420086 TI - Epidermoid cysts of testes and role of sonography. AB - Four patients with histologically proved epidermoid cysts of the testicle were managed with radical orchiectomy. High-resolution testicular sonography demonstrated similar findings in all cases, but these findings were not specific and did not preclude other malignant or teratomatous lesions. The lack of both unique clinical features and specific sonographic findings would suggest that extreme caution must be used if wedge resection or enucleation is contemplated for a patient with presumed epidermoid cyst. PMID- 8420087 TI - Contrast media reactions during voiding cystourethrography or retrograde pyelography. AB - Contrast media can be absorbed across the urothelium in amounts sufficient to cause severe anaphylactoid reactions. We report 2 cases of anaphylactoid reactions during voiding cystourethrography (VCUG) or retrograde pyelography (RGP). A retrospective review of 783 consecutive VCUGs or RGPs performed at our institution over the last five years revealed no other systemic contrast media reactions. Although our findings and a review of the literature discloses that the incidence appears to be extremely low, urologists and radiologists need to be aware that an anaphylactoid or vagal contrast medium reaction may develop during VCUGs or RGPs. In addition, these physicians need to be prepared to treat a reaction should one occur. PMID- 8420088 TI - Twenty-one-year-old woman with flank pain. PMID- 8420089 TI - Enhanced intravenous urography in renal trauma with low-dose dopamine. AB - Intravenous pyelography is the standard first-line investigation for suspected renal trauma. A faint, and/or delayed visualization, or nonvisualization of the damaged renal unit is not uncommon. Low-dose dopamine (3 micrograms/kg/min) increases renal blood flow without deleterious side effects. An experimental rat model was developed to evaluate the effects of low-dose dopamine on intravenous pyelograms in animals with unilateral renal trauma. A consistent and significant improvement in the visualization of the injured kidney was noted in the dopamine treated animals compared with controls that received equivalent volumes of normal saline. PMID- 8420090 TI - Role of lymphadenectomy in the treatment of renal cell carcinoma. AB - We evaluated the role of lymphadenectomy (LND) in the prevention of local recurrence following radical nephrectomy for renal cell carcinoma (RCC) by two retrospective studies. In one, the relative importance of various tumor characteristics to the subsequent development of local recurrence was investigated in 37 patients who underwent radical nephrectomy and later progressed. In 29 evaluable patients, only nodal metastasis was predictive of local recurrence, which developed in 6 of 7 node-positive patients. In our second study the records of 69 consecutive patients with RCC who underwent radical nephrectomy with or without simultaneous LND (N = 42 and 27, respectively) were reviewed. Local control after LND was excellent in node-positive disease; in no node-positive patient with unilateral RCC has a local recurrence developed (N = 5). LND did not extend hospitalization or add to the morbidity of radical nephrectomy. PMID- 8420091 TI - Endometrioid adenocarcinoma of prostatic urethra presenting with anterior urethral implantation. AB - Endometrioid adenocarcinoma of the prostatic urethra is presented in an eighty four-year-old white man who had been treated for transitional cell carcinoma of bladder and microacinar adenocarcinoma of prostate prior to presenting with an implanted anterior urethral mass. The differential diagnosis of the papillary lesions encountered in the prostatic and penile urethra is reviewed. Based on the literature, a comparative study between both pure and mixed endometrioid carcinoma and microacinar prostatic carcinoma was done. The study aimed to elicit more data about the clinical features of endometrioid carcinoma and included several clinical parameters (age, presenting symptoms, rectal examination findings, serum acid phosphatase level, and metastasis at presentation). The general impression gained from this study is that no significant clinical difference exists between these two histologic variants of prostatic carcinoma. PMID- 8420092 TI - Vesicolevator reflex. Description of a new reflex and its clinical significance. AB - A new reflex which I call "vesicolevator reflex" was studied in 21 healthy volunteers with a mean age of 36.3 years. The technique comprised the introduction of a catheter, with a balloon at its distal part, into the empty urinary bladder. A concentric needle electrode was inserted into the levator ani muscle. The vesical balloon was inflated with air in increments of 50 mL and the levator myoelectric activity recorded by a standard EMG apparatus. Vesical balloon was then removed, and levator EMG response to sudden suprapubic vesical compression was recorded with urinary bladder both empty and filled with saline in increments of 50 mL. Vesical balloon distention evoked levator muscle contraction; duration of contraction increased with increased vesical distention. Levator response to vesical distention did not occur when air-filling was below 50 mL air and when muscle was anesthetized. The latency of the reflex was calculated. Suprapubic manual compression of the empty urinary bladder did not cause levator contraction; yet, compression of distended bladder evoked the response. Compression should be sudden, while the patient is lying supine. Slow insidious compression did not evoke levator contraction. The vesicolevator reflex seems to play a role in the act of micturition, and hence could serve as a tool in investigating patients with micturition disorders. PMID- 8420093 TI - Endocavitary bladder surgery. AB - The advent of laparoscopy has expanded the horizon for endocavitarily approaching urologic disorders, otherwise managed by open surgical procedures. In this article we will review our experience with this new modality as applied to the urinary bladder, and put into perspective future applications. PMID- 8420094 TI - Balloon expanded titanium prostatic urethral stent. AB - A new balloon expanded titanium prostatic urethral stent has been devised for the treatment of outflow obstruction. This stent is inserted under endoscopic guidance and endourethral anesthesia. We have treated 9 male patients in urinary retention with a mean follow-up of eighteen months. Six of our 9 patients had a good and prolonged response to stenting. Complications were minimal and the procedure was well tolerated. We believe that the balloon expandable titanium stent is a viable alternative for the treatment of urinary retention particularly in the very ill patient. PMID- 8420095 TI - Endoscopic removal of lower ureter in nephroureterectomy. AB - Endoscopic removal of the lower ureteral stump by transurethral resection of the mucosal cuff and intussusception of the ureter is safe, effective, and relatively easy. Although the technique has been used primarily for malignant disease, the cases described herein illustrate that it also is useful for benign disease. PMID- 8420096 TI - Use of transrectal ultrasound in follow-up of postradical prostatectomy. AB - The appearance of the prostatic fossa on transrectal ultrasound following radical retropubic prostatectomy (RRP) is described. Transrectal ultrasonography was performed on 25 patients with normal bone scans and pelvic computed tomography from three to ninety months after RRP using a biplane high frequency probe. The area of the vesicourethral anastomosis (VUA) was identified, its contour characterized, surrounding tissues described, and changes induced by pelvic muscle contraction recorded. In 16 of these patients who had abnormal postoperative serum prostate-specific antigen levels, digital and ultrasound guided transrectal needle biopsies for local recurrence were done and compared. The VUA was identifiable in all patients as either a smoothly tapered narrowing usually correlating with the presence of continence or distorted or blunted profile which often correlated with absence of urinary continence. The VUA was surrounded almost invariably by hypoechoic soft tissue which was pathologically nonspecific on biopsy. An extrinsic impression on the anterior bladder wall was noted in 80 percent. There was no clear distinguishing ultrasound feature for biopsy-proved local recurrence. The apparent length of the apposed walls of the urethra suggests a urethral high pressure zone (UHPZ). This lengthened significantly with voluntary contraction of the pelvic floor muscles. PMID- 8420098 TI - Regeneration in Alzheimer disease and aging. PMID- 8420097 TI - Prolongation of response to DMSO by heparin maintenance. AB - Dimethylsulfoxide (DMSO) is an effective treatment of symptomatic patients with detrusor mastocytosis but it is associated with frequent relapses. A group of patients (N = 25) followed for twelve months showed a relapse rate of 59 percent. Our experience with a combination of DMSO and heparin has suggested that the relapse rate may be lower. Heparin is a glycosaminoglycan that may afford protection to the urothelium and may reduce the relapse rate. It is better tolerated than DMSO or a combination of DMSO and heparin and does not produce garlic halitus. It is not associated with coagulation anomalies when administered intravesically. To determine whether or not maintenance therapy with intravesical heparin may reduce relapses we have treated a similar cohort of 25 patients with monthly instillations of 10,000 IU of heparin over a twelve-month period. Both groups were comparable in age, duration of symptoms, severity of symptoms, and response to DMSO. At twelve months only 20 percent of the heparin-treated group had relapsed versus 52 percent of the control group. Furthermore 6 patients (24%) in the follow-up group failed to respond to retreatment with DMSO while all of the heparin maintenance group continued to respond to one or more treatments with DMSO. Thus, it seems that heparin maintenance produces a significant reduction in the relapse rate of patients who respond to DMSO and reduces the number of patients requiring alternative therapy. PMID- 8420099 TI - Essential prerequisites for remyelination of oligodendrocytes. Division, motility, and structural rearrangement. PMID- 8420100 TI - Source of remyelinating oligodendrocytes. PMID- 8420101 TI - The role of astrocytes in the remyelination of glia-free areas of demyelination. PMID- 8420102 TI - Spinal cord injury: physiology and transplantation. PMID- 8420103 TI - Adaptive plasticity in spinal cord. PMID- 8420104 TI - Inhibitory control of reflex excitability following contusion injury and neural tissue transplantation. PMID- 8420105 TI - Intraspinal transplants of catecholamine-containing cells and fetal spinal cord and iris tissues in the adult rat. PMID- 8420106 TI - Steroid hormones and neuronal regeneration. PMID- 8420107 TI - Gonadal steroids and neuronal regeneration. A therapeutic role. PMID- 8420108 TI - The effects of glucocorticoid and nonglucocorticoid steroids on acute neuronal degeneration. AB - The glucocorticoid steroid methylprednisolone (MP) has been shown to enhance chronic recovery after human spinal cord injury when administered in a 24-hr high dose regimen beginning within 8 hr. The doses of MP that effect this improved recovery have been demonstrated to inhibit spinal tissue LP, which has been postulated to be a key event in the secondary posttraumatic degenerative cascade. The molecular mechanism of action of the steroid appears to involve intercalation into the cell membrane and blockade of the propagation of LP reactions. At a physiological level, the inhibition of injury-induced LP has been found to result in an attenuation of progressive posttraumatic ischemia and energy failure together with an augmented reversal of intracellular calcium accumulation. However, MP also acts directly to retard secondary neuronal degeneration, as observed in studies showing the steroid's ability to slow the anterograde degeneration of experimentally injured cat soleus motor nerves. The duplication of this effect by the nonsteroidal lipid antioxidant alpha-tocopherol supports the notion that it is indeed a manifestation of the inhibition of posttraumatic LP. Moreover, the efficacy of MP in limiting lipid peroxidation and secondary spinal cord or motor nerve degeneration has also been duplicated by a nonglucocorticoid 21-aminosteroid, tirilazad mesylate (U-74006F), which suggests the independence of the antioxidant and glucocorticoid effects of MP. PMID- 8420109 TI - Strategies for the development of new and better pharmacological treatments for acute spinal cord injury. PMID- 8420110 TI - Regulatory genes in regeneration and plasticity. PMID- 8420111 TI - The role of immediate early genes in the regeneration of the central nervous system. PMID- 8420112 TI - Early response genes as markers of neuronal activity and growth factor action. PMID- 8420113 TI - A fos-lac Z transgenic mouse that can be used for neuroanatomic mapping. AB - Cellular immediate-early genes are rapidly induced by a diverse range of agents and conditions. Since many cIE genes encode known or potential transcription factors, they are believed to couple extracellular stimuli to long-lasting alterations in cellular phenotype through the regulation of gene transcription. In addition, the localization of the products of cIE genes has been used as a method for determining the cellular sites of action of particular agents in the nervous system. However, the methods of analysis are tedious, and the results may be ambiguous because of cross-reaction of reagents with related proteins. To further the utility of this approach, a bacterial gene encoding beta galactosidase (lac Z) has been fused, in frame, into the fourth exon of c-fos, and this fos-lac Z fusion gene has been introduced into the germ line of mice. We have analyzed the expression of beta-galactosidase (under the control of the c fos promoter) in the developing and adult nervous systems of these transgenic mice. As far as can be determined, the constitutive and stimulated expression of the transgene accurately reflects the expression of cognate c-fos in both cultured cells and the intact animal. This study has also revealed novel sites of constitutive and induced expression of c-fos that were overlooked using conventional analysis. In particular, constitutive expression of c-fos is associated with cells that are entering terminal differentiation and are destined to die. In addition, induced expression of the transgene in adult brain mirrors the pattern of neurotoxicity elicited by kainic acid.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420114 TI - Stress (heat shock) protein accumulation in the central nervous system. Its relationship to cell stress and damage. PMID- 8420115 TI - The function of phagocytic cells in the nervous system. PMID- 8420116 TI - Macrophage responses to central and peripheral nerve injury. PMID- 8420117 TI - Microglial secretion products and their impact on the nervous system. PMID- 8420118 TI - Macrophage functional properties during myelin degradation. PMID- 8420119 TI - Novel major histocompatibility complex expression by microglia and site-specific experimental allergic encephalomyelitis lesions in the rat central nervous system after optic nerve transection. PMID- 8420120 TI - Expression of monocyte/macrophage growth factors and receptors in the central nervous system. PMID- 8420121 TI - Adaptive versus pathological plasticity. Possible contributions to age-related dementia. PMID- 8420122 TI - Connections among plasticity, regeneration, and aging at the neuromuscular junction. PMID- 8420123 TI - The histopathology of neuronal degeneration and plasticity in Alzheimer disease. PMID- 8420124 TI - The role and repair of forebrain cholinergic systems in short-term memory. Studies using the delayed matching-to-position task in rats. PMID- 8420125 TI - Myelin injury and repair. PMID- 8420126 TI - Observations on the pathology of human spinal cord injury. A review and classification of 22 new cases with details from a case of chronic cord compression with extensive focal demyelination. PMID- 8420127 TI - Remyelination, revascularization, and recovery of function in experimental spinal cord injury. PMID- 8420128 TI - Parkinson's disease. The L-dopa era. PMID- 8420129 TI - Selective D1 or D2 receptor blockade. Effects on brain functional responses to L dopa. PMID- 8420130 TI - Transferrin receptor regulation in Parkinson's disease and MPTP-treated mice. PMID- 8420131 TI - Parkinson's disease and the adaptive capacity of the nigrostriatal dopamine system: possible neurochemical mechanisms. PMID- 8420132 TI - Are dopaminergic neurons selectively vulnerable to Parkinson's disease? PMID- 8420133 TI - Striatal dopaminoceptive system changes and motor response complications in L dopa-treated patients with advanced Parkinson's disease. PMID- 8420134 TI - Metabolic aspects of the behavior of MPTP and some analogues. PMID- 8420135 TI - N-methylation of pyridines and Parkinson's disease. PMID- 8420136 TI - MPTP and aging. Studies in the C57BL/6 mouse. PMID- 8420137 TI - N-methylated tetrahydroisoquinolines as dopaminergic neurotoxins. AB - N-Methylation of dopamine-derived 6,7-dihydroxyisoquinolines produces compounds whose chemical structures are similar to 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). N-Methylation of 1,2,3,4-tetrahydroisoquinoline was proved by in vitro experiments using human brain homogenate as an enzyme source. By in vivo microdialysis in rat brains, N-methylation of 1-methyl-6,7-dihydroxy 1,2,3,4-tetrahydroisoquinolines (salsolinols) was also demonstrated. Among the brain regions examined, the activity of N-methylation was the highest in the substantia nigra. N-methyl-1,2,3,4-tetrahydroisoquinoline was oxidized to produce N-methyl-isoquinolinium ion by type A and type B monoamine oxidase prepared from human brain synaptosomal mitochondria. The structure of the oxidized isoquinoline is similar to that of 1-methyl-4-phenylpyridinium ion, a potent dopaminergic neurotoxin. These results indicate that these isoquinolines produced in or around dopamine neurons are N-methylated especially in substantia nigra and may be further oxidized by monoamine oxidase to produce ions. This biosynthesis pathway is quite similar to the oxidative synthesis of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine. N-Methylation and oxidation of isoquinolines or other endogenous compounds may be involved in the pathogenesis of Parkinson's disease. PMID- 8420138 TI - Dopamine-derived tetrahydroisoquinolines and Parkinson's disease. AB - The daily urinary excretion of salsolinol, 1,2-dehydrosalsolinol, and norsalsolinol, as free, glucuronide, and sulfate, has been measured in parkinsonian patients and age-matched controls in an attempt to examine whether the determination of dopamine-derived alkaloids in urine may be used as a marker of the decrease in brain dopamine levels associated with the disease. In contrast with a preliminary study where the daily urinary excretion of total salsolinol was significantly higher in young controls than in parkinsonians, in the present study no difference was found between parkinsonian patients and controls concerning salsolinol and norsalsolinol excretion. However, the urinary excretion of total 1,2-dehydrosalsolinol was significantly higher in the control group, owing to a statistically significant increase in its excretion as sulfate in this group. Further studies appear to be necessary to establish whether 1,2 dehydrosalsolinol, salsolinol, and/or any other dopamine-derived alkaloid may serve for the detection of subjects with dysfunctions of the dopaminergic system. PMID- 8420139 TI - Biosynthesis of 1-methyl-1,2,3,4-tetrahydroisoquinoline (1MeTIQ), a possible antiparkinsonism agent. PMID- 8420140 TI - Presence of tetrahydroisoquinoline-related compounds, possible MPTP-like neurotoxins, in parkinsonian brain. PMID- 8420141 TI - Monoamine oxidase-B, monoamine oxidase-B inhibitors, and Parkinson's disease. A role for superoxide dismutase? PMID- 8420142 TI - Iron and ferritin in substantia nigra in Parkinson's disease. PMID- 8420143 TI - Implications of alterations in trace element levels in brain in Parkinson's disease and other neurological disorders affecting the basal ganglia. PMID- 8420144 TI - Mitochondrial energy crisis in Parkinson's disease. PMID- 8420145 TI - Mitochondrial complex I deficiency in Parkinson's disease. PMID- 8420146 TI - The genetics of Parkinson's disease. A review. PMID- 8420147 TI - Debrisoquine hydroxylase and Parkinson's disease. AB - The relationship between the genotypes of Xba I and Bam H I restriction fragment length polymorphisms (RFLPs) at a gene for debrisoquine hydroxylase (CYP2D6) and phenotypes of the metabolic function of debrisoquine/sparteine, the EM and the PM in a healthy Japanese population was investigated. The genotypes of Xba I 11.5 kb and Xba I 44 kb-Bam H I 2.3 kb- were responsible for the Japanese PM. Genotype of Xba I RFLP at CYP2D6 locus was analyzed in 43 healthy individuals and 51 patients with IDP. The relative risk of IDP was 2.15 times more for individuals with the Xba I 44 kb allele compared to those without the allele (chi 2 = 4.149, d.f. = 1, p < 0.05) and it was 6.32 times greater for the Xba I 44 kb homozygotes than the Xba I 29 kb homozygotes (chi 2 = 4.935, d.f. = 1, p < 0.05). These data suggest that the PM for debrisoquine/sparteine hydroxylase might be one of the genetic factors making humans susceptible to IDP acquisition. PMID- 8420148 TI - Early-stage loss of dopamine uptake-site binding in MPTP-treated monkeys. PMID- 8420149 TI - Promoter structure of the human gene coding for the D1A dopamine receptor. PMID- 8420150 TI - Lifestyles, risk factors, and inherited predispositions in Parkinson's disease. Preliminary report of a case-control study. PMID- 8420151 TI - Epidemiological study of Parkinson's disease in Parsis in India. PMID- 8420152 TI - Circulating antibody to Nocardia in the serum of patients with Parkinson's disease. AB - Nocardia is a genus of aerobic Gram-positive bacteria which forms filamentous cells that fragmented into rod-shaped or coccoid elements or L-forms. According to the literature N. asteroides causes encephalitis with parkinsonian features, and it may be an unknown etiologic agent causing encephalitis with a parkinsonian syndrome. Because there are no reliable immunologic tests routinely used for diagnosing nocardial infection, less severe CNS infections with nocardiae may not be recognized or may be attributed to unknown or incorrect etiologic agents. Recently, it was shown that sublethal doses of N. asteroides causes an L-DOPA responsive movement disorder with Lewy-like bodies in mice. In this study, we detected antibodies to nocardia in the serum of patients with Parkinson's disease (PD). Sera was diluted in PBS and added in two-fold dilutions to coccoid and rod shaped cells of nocardia attached to nonfluorescence slides. IF test demonstrated antibodies to coccoid and rod-shaped cells of nocardia in the serum from 20/20 patients with PD at a titer greater than 1:10, and 14 controls showed 10 positively. The results suggested that not only PD patients but also age-matched healthy volunteers are routinely exposed to and naturally infected with nocardia related microorganisms. A further reliable immunologic test will be required. PMID- 8420153 TI - Physiological mechanisms and assessment of motor disorders in Parkinson's disease. PMID- 8420154 TI - Sensorimotor processing in Parkinson's disease. PMID- 8420155 TI - Does abnormal stimulus processing contribute to delayed motor response in Parkinson's disease? PMID- 8420156 TI - Reflex behavior and programming in Parkinson's disease. AB - Several aspects of characteristically impaired postural adjustments indicate a defective central programming of the respective EMG patterns in PD. This may be due to deficits in the supraspinal control of spinal interneuronal circuits. This impairment obviously contributes to the difficulty of these patients in performing two motor acts simultaneously. In addition, parkinsonian patients exhibit a reduced sensitivity of polysynaptic reflexes in the leg extensor muscles which correlates with their postural instability. The activity of monosynaptic reflexes is negligible, as in healthy subjects. The impairment of proprioceptive reflex function may be partially compensated for by changes of intrinsic muscle stiffness. Discrepancies in the literature about the behavior of the polysynaptic EMG responses in parkinsonian patients may in part arise from the fact that it depends on both the particular muscle under study and the actual motor task investigated. PMID- 8420157 TI - Contribution of reticular nuclei to the pathophysiology of parkinsonian rigidity. PMID- 8420158 TI - Early detection of L-dopa response in parkinsonian patients with a standardized tracking test. PMID- 8420159 TI - The visual system in Parkinson's disease. PMID- 8420160 TI - Vestibulo-ocular reflex in Parkinson's disease and multiple system atrophy. PMID- 8420161 TI - Bereitschaftspotential preceding voluntary saccades is abnormal in patients with Parkinson's disease. PMID- 8420162 TI - Motor performance in parkinsonism following stereotactic thalamotomy. PMID- 8420163 TI - Long-term effects of chronic stimulation of the ventral intermediate thalamic nucleus in different types of tremor. PMID- 8420164 TI - Gait disorders in parkinsonism. A study with floor reaction forces and EMG. PMID- 8420165 TI - Impaired activity of the supplementary motor area in akinetic patients with Parkinson's disease. Improvement by the dopamine agonist apomorphine. PMID- 8420166 TI - Spatiotemporal study of event-related desynchronization in idiopathic Parkinson's disease. PMID- 8420167 TI - Event-related potentials, reaction time, and cognitive state in patients with Parkinson's disease. AB - ERPs, RT, and cognitive state in 53 patients with PD were studied. The peak latency of P300 and the RT in PD patients were significantly prolonged in the advanced stage of illness. The cognitive state in PD patients was characterized by impairment of attention, recent memory, and constructional ability in categories of the Folstein Mini-Mental State Examination. The significant association between neurophysiological and neuropsychological measurements suggests that these measurements reflect a disrupted aspect of cognitive function in PD patients. PMID- 8420168 TI - Urodynamic study in the differential diagnosis between multiple system atrophy and Parkinson's disease. PMID- 8420169 TI - Dementia in Parkinson's disease. Morphometric data. PMID- 8420170 TI - Dementia and neuropathology in Lewy body disease. AB - In order to elucidate the pathological differences between demented and nondemented patients with Lewy body disease, brains from 35 patients were clinicopathologically examined. In diffuse Lewy body disease, cortical lesions, including numerous Lewy bodies and senile changes, were found to be responsible for the dementia. In some of the cases with the brain-stem type of Lewy body disease, the dementia was attributed to an Alzheimer pathology, while in many cases the various combinations of degeneration in the subcortical nuclei, mainly the nucleus basalis of Meynert and locus ceruleus, played a major role in the dementia. Forty-four Japanese cases with diffuse Lewy body disease were reviewed. Then diffuse Lewy body disease was divided into two forms: a common form (33 cases) and a pure form (11 cases). In the common form, all cases showed progressive cortical dementia in the presenile or senile period. Parkinson syndrome was usually marked in the terminal stage. However, about a fifth of the cases had no parkinsonism. Neuropathologically, the common form had many concomitant senile changes in the cerebral cortex. Most cases with the pure form showed juvenile Parkinson syndrome followed by progressive cortical dementia, while a few cases were of presenile or senile occurrence. Neuropathologically, the pure form had no or few senile changes. This suggests that numerous cortical Lewy bodies alone can cause cortical dementia. PMID- 8420171 TI - Clinical and neuropathological aspects of diffuse Lewy body disease in the elderly. AB - This chapter reports the clinical and neuropathological findings of eight cases of "diffuse Lewy body disease" verified by autopsy. The age at onset was between 60 and 82 years; the age at death was between 75 and 92 years. The initial symptoms were amnesia in three cases, orthostatic dizziness in three, visual hallucination in two, but parkinsonism in none. The cardinal clinical symptoms included dementia in all cases, hallucinatory-delusional state in six, akinesia and rigidity in five, and orthostatic hypotension in five. Antemortem diagnoses were senile dementia in five, and hallucinatory-delusional state, Parkinson's disease and Shy-Drager syndrome in one each. Despite the clinical symptoms differences from each other, neuropathological findings were alike. Abundant Lewy bodies were present in the neurons of the cerebral cortex as well as in the brainstem nuclei and diencephalon. Concomitant senile changes including senile plaques and Alzheimer's neurofibrillary tangles (NFTs) were also present in varying degree. Immunocytochemical study with anti-ubiquitin for Lewy body, anti tau protein for NFT, and beta-protein of amyloid for senile plaque suggested that dementia of DLBD might have resulted not from a single pathology but from the complex of Lewy bodies, NFTs and senile plaques. PMID- 8420173 TI - L-dopa and frontal cognitive function in Parkinson's disease. PMID- 8420172 TI - A controlled study of dementia in Parkinson's disease over 54 months. PMID- 8420174 TI - Chronic dementia in Parkinson's disease treated by anticholinergic agents. Neuropsychological and neuroradiological examination. AB - Reports about chronic-onset dementia due to ACA have been rare. Here we described five patients with chronic dementia induced by ACA. We examined their clinical aspect, neuropsychological aspect, and neuroradiological aspect in detail. Our study suggests the possibility of ACA-induced chronic-onset dementia in PD patients. PMID- 8420175 TI - PET imaging in Parkinsonism. PMID- 8420176 TI - Dopamine D1 and D2 receptors in Parkinson's disease and striatonigral degeneration determined by PET. PMID- 8420178 TI - Positron emission tomography in hemiparkinsonism-hemiatrophy syndrome. PMID- 8420177 TI - Dopamine D2 receptor imaging and measurement with SPECT. AB - Only recently it has become possible to label and visualize dopamine D2 receptors in the living human brain using an [123I]labeled ligand and the SPECT technique. Iodobenzamide (S(-)IBZM) is a substance with high affinity and high specificity for D2 receptors and was used in controls and in patients with different extrapyramidal disorders. After the i.v. administration of 5mCi (185 MBq) of [123I] labeled S(-)IBZM data collection was performed with a rotating double head scintillation camera between 60 and 110 minutes. In a semiquantitative approach a ratio was calculated between mean counts/pixel in the striatum and a region in the lateral frontal cortex, which gives an index of receptor density. In a group of 21 controls this ratio is 1.73 +/- 0.09. A highly significant age-related decline is found in controls and in 57 patients with PD. PD patients without L DOPA or dopamine agonist treatment do not differ from controls (1.72 +/- 0.09; n = 18), whereas patients under dopaminergic therapy show a significantly lower binding ratio (1.65 +/- 0.13; n = 39; p = 0.017) suggesting receptor downregulation. Comparing the striatum ipsi- and contralateral to clinical symptoms in 18 hemiparkinsonian patients shows slightly but significantly higher values on the contralateral side, indicating receptor supersensitivity (1.71 +/- 0.11 vs. 1.66 +/- 0.09; p = 0.0014). No correlation between D2 receptor binding and clinical stage, duration of disease or duration of dopaminergic therapy exists. Markedly reduced ratios are measured in 7 patients with progressive supranuclear palsy and multiple system atrophies and in 18 patients with Huntington's disease (1.40 +/- 0.09 and 1.37 +/- 0.12 respectively; p = 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420179 TI - Impaired activation of frontal areas during movement in Parkinson's disease: a PET study. PMID- 8420180 TI - Different cerebral metabolism between parkinsonian rigidity and hyperkinesia (DID, chorea, dystonia). A PET study. PMID- 8420181 TI - Conversion of alpha-methyltryptophan to alpha-methylserotonin in vivo. New techniques for imaging serotonin. PMID- 8420182 TI - IBZM-SPECT as predictor for dopamimetic responsiveness of patients with de novo parkinsonian syndrome. AB - IBZM-SPECT is useful as a predictor for the responsiveness to oral L-DOPA therapy in de novo parkinsonian patients. A reduction of postsynaptic dopamine D2 receptors in de novo parkinsonian patients makes the diagnosis of Parkinson's disease rather unlikely. Normal IBZM binding does not prove the diagnosis of Parkinson's disease; however, together with a positive apomorphine test, it strongly supports the clinical diagnosis. At present the combination of IBZM SPECT and apomorphine testing appears to be a useful procedure for selecting de novo patients for clinical trials with new anti-parkinsonian therapy. PMID- 8420183 TI - IBZM-SPECT imaging in Parkinson's disease. Quantification of binding ratios from sequential SPECT measurements in patients and controls. PMID- 8420184 TI - Magnetic resonance imaging in parkinsonisms. PMID- 8420185 TI - Pathophysiology of parkinsonian motor abnormalities. PMID- 8420186 TI - MRI pathology of basal ganglia in dystonic disorders. PMID- 8420187 TI - Parkinsonism and dystonia, pseudo-parkinsonism and pseudodystonia. PMID- 8420188 TI - Assessment of the dopaminergic lesion in Parkinson's disease by CSF markers. PMID- 8420189 TI - Nosological concept of juvenile parkinsonism with reference to the dopa responsive syndrome. PMID- 8420190 TI - Parkinson's disease before age 30. PMID- 8420191 TI - Dystonia and Parkinson's disease. PMID- 8420192 TI - CSF biopterin levels and clinical features of patients with juvenile parkinsonism. PMID- 8420193 TI - Hereditary progressive dystonia with marked diurnal fluctuation. Pathophysiological importance of the age of onset. PMID- 8420194 TI - Dopa-responsive dystonia. Delineation of the clinical syndrome and clues to pathogenesis. PMID- 8420195 TI - Clinical heterogeneity of dopa-responsive dystonia: PET observations. AB - Parkinsonism without dystonia has been reported in several older members of families with DRD. This raises the question whether such patients represent a variant in the clinical picture of DRD, or a separate disease, IP. We employed 6 FD PET to study the nigrostriatal dopaminergic function in a woman with typical DRD and two of her relatives with late-onset parkinsonism. These two had an excellent and prolonged therapeutic response to small doses of L-DOPA, without complications. We found that the dystonic patient and the women with "benign" parkinsonism had normal striatal 6-FD uptake. In conjunction with other clinical evidence, our PET study indicates that the biochemical lesion in these members may be "proximal" to dopa decarboxylase, as is suggested in DRD patients. We conclude that the adult-onset parkinsonism in DRD families is due to the same pathophysiological mechanism as the childhood-onset dystonia in the disease. DRD may display substantial clinical heterogeneity depending on the age of onset. PMID- 8420196 TI - PET scan study on the dopaminergic system in a Japanese patient with hereditary progressive dystonia (Segawa's disease). Case report. PMID- 8420197 TI - The clinical features of Parkinson's disease in 100 histologically proven cases. PMID- 8420198 TI - Lewy bodies in the visceral autonomic nervous system in Parkinson's disease. PMID- 8420199 TI - Familial parkinsonism and dementia with "ballooned neurons". PMID- 8420200 TI - Progressive supranuclear palsy presenting with pure akinesia. PMID- 8420201 TI - Dopa-unresponsive pure akinesia or freezing. A condition within a wide spectrum of PSP? PMID- 8420202 TI - Future strategies for the treatment of Parkinson's disease. PMID- 8420203 TI - Potent COMT inhibition by Ro 40-7592 in the periphery and in the brain. Preclinical and clinical findings. PMID- 8420204 TI - Superior clinical efficacy of Sinement CR 50/200 versus Sinemet 25/100 in patients with fluctuating Parkinson's disease. An open and a double-blind, double dummy, multicenter treatment evaluation. Dutch Sinement CR Study Group. PMID- 8420205 TI - Apomorphine and lisuride infusion. A comparative chronic study. PMID- 8420206 TI - Continuous subcutaneous apomorphine infusions for fluctuating Parkinson's disease. Long-term follow-up in 18 patients. PMID- 8420207 TI - MAO-B inhibitors in Parkinson's disease. PMID- 8420208 TI - Protective effect of selegiline in the early and late phases of Parkinson's disease. PMID- 8420209 TI - An open multicenter study of the efficacy of MDL 72,974A, a monoamine oxidase type B (MAO-B) inhibitor, in Parkinson's disease. PMID- 8420210 TI - Broad bean (Vicia faba) consumption and Parkinson's disease. AB - In 1913, Guggenheim identified L-DOPA in the seedlings, pods, and beans of the broad bean, Vicia faba (VF). Since then, anecdotal cases of symptomatic improvement after VF consumption have been described in patients with PD. In the present study, five healthy volunteers and six PD patients (mean age, 63.5 years; mean disease duration, 13 years; stage III, Hoehn-Yahr scale) ate 250 g cooked VF after 12 hours off medication. Blood samples for L-DOPA measurements (by HPLC-ED) were obtained before eating VF and every 30 minutes, for 4 hours. During this period, a substantial clinical improvement was noted and three patients also showed severe dyskinesias. High plasma L-DOPA values were also measured (Cmax 0.66 to 1.20 micrograms/ml; AUC 1.82 to 4.12 micrograms/ml/H). In addition, their clinical performance and plasma L-DOPA levels were compared to those found after 125 mg L-DOPA + 12.5 mg carbidopa ingested on another day. These data show that VF ingestion produces a substantial increase in L-DOPA plasma levels, which correlates with a substantial improvement in motor performance. Our findings may have implications for the treatment of PD, especially in patients with mild symptoms. PMID- 8420211 TI - Is the decreasing responsiveness of axial motor symptoms to L-dopa in elderly parkinsonian patients related to age-associated nonparkinsonian pathologies? PMID- 8420212 TI - Clinical features and medical treatment of Parkinson's disease in patient groups selected in accordance with age at onset. AB - In a recent hospital-based study on patients with different age at onset of PD, some clinical and therapeutic features were investigated. In the retrospective analysis three groups were selected: age at onset between 20 and 39 years (young onset), between 40 and 59 years, and from 60 years and up. The evaluation of our data demonstrates some significant differences in the above-mentioned groups: prevalence of male patients and more frequent occurrence of motor fluctuations and dyskinesias in the young onset group, longer duration of PD and L-DOPA treatment, higher L-DOPA dosage, and more multiple combinations in this group. We also noted more habitual smokers in the young onset group. Sex-specific differences in the young onset group were also investigated. A longer duration of disease was found in women, and some symptoms, such as pulsion and freezing and biphasic dyskinesia, are more frequent in women. PMID- 8420213 TI - Risk factors for motor response complications in L-dopa-treated parkinsonian patients. PMID- 8420214 TI - Treatment of drug-induced psychosis in Parkinson's disease with clozapine. PMID- 8420215 TI - Tissue implants in treatment of parkinsonian syndromes in animals and implications for use of tissue implants in humans. PMID- 8420216 TI - [18F]-6-L-fluorodopa PET studies of graft and host dopaminergic function following fetal mesencephalic transplantation. PMID- 8420217 TI - Embryonic dopamine cell implants as a treatment for the second phase of Parkinson's disease. Replacing failed nerve terminals. PMID- 8420218 TI - Fifteen months' follow-up on bilateral embryonic mesencephalic grafts in two cases of severe MPTP-induced parkinsonism. PMID- 8420219 TI - Characterization of brain microglia and the biological significance in the central nervous system. PMID- 8420220 TI - Dynamic aspects of the striatothalamic connection studied in cases with movement disorder. PMID- 8420221 TI - The role of the medial pallidum in the pathophysiology of akinesia in primates. PMID- 8420222 TI - The role of basal ganglia in rhythmic movement. PMID- 8420223 TI - Patency of the infarct-related artery and left ventricular function as the major determinants of survival after Q-wave acute myocardial infarction. AB - One hundred seventy-two patients with 1-vessel disease documented at predischarge angiography who had been followed for 43 +/- 30 months after an initial Q-wave acute myocardial infarction were retrospectively evaluated to investigate the prognostic value of infarct-related artery patency and left ventricular (LV) function. Multiple logistic regression analysis revealed that only infarct artery patency (Thrombolysis in Myocardial Infarction [TIMI] grades 2-3 vs 0-1) (Z = 2.24; p < 0.05) and end-systolic volume index (Z = -2.67; p < 0.01) were independently related to survival. Sixteen cardiac deaths were observed; all 16 patients had LV dysfunction (defined as end-systolic volume index > 40 ml/m2), and 15 had an occluded infarct-related artery. In the subgroup with LV dysfunction, the 10-year percent survival rate was 20% among patients with TIMI grade 0 to 1 versus 96% with grade 2-3 (p < 0.001). Patency of the infarct related artery was also the only independent predictor of recurrent ischemia (Z = 2.59; p < 0.01). In conclusion, both infarct-related artery patency and LV function are independent predictors of survival after Q-wave acute myocardial infarction. Patients with normal LV function have an excellent long-term prognosis, which is only partially counterbalanced by the tendency toward clinical instability observed in those with an open infarct-related vessel. However, when an occluded infarct-related artery is observed in the setting of LV dysfunction, the long-term outcome appears to be relatively poor. PMID- 8420224 TI - Measurement of cardiac output by automated single-breath technique, and comparison with thermodilution and Fick methods in patients with cardiac disease. AB - Accurate noninvasive methods are needed for determination of cardiac output. Current methods are generally complex or may be unreliable. A previously described method, based on absorption of acetylene gas during a constant exhalation that enables calculation of cardiac output by estimating pulmonary capillary circulation, is incorporated in a new, automated commercial product (SensorMedics 2200). In this study, cardiac output by single-breath acetylene blood flow measured with this device was compared with the standard thermodilution and direct Fick methods in 20 patients undergoing cardiac or pulmonary artery catheterization. Patients inhaled test gas mixture to total lung capacity and exhaled at a constant rate through an adjustable resistor. Lung volumes and noninvasive acetylene blood flow value were calculated automatically. Correlation between the automated single-breath technique and both thermodilution and Fick cardiac output determinations was very high (correlation coefficients were 0.90 and 0.92, respectively), regression slopes were close to identity (0.98 and 0.90), and bias (-0.39 and -0.79 liter/min) and precision (0.94 and 1.02) were good; when shunt correction was applied, bias was reduced to 0.06 and 0.35 liter/min, respectively. Rapid, accurate, noninvasive measurement of cardiac output was easily obtained using the automated device. This technique may have a wide applicability for noninvasive evaluation of patients with cardiac disease and for monitoring effects of therapeutic interventions. PMID- 8420225 TI - Prognostic significance of silent myocardial ischemia in patients > 61 years of age with extracranial internal or common carotid arterial disease with and without previous myocardial infarction. PMID- 8420226 TI - Ventricular fibrillation markers on admission to the hospital for acute myocardial infarction. PMID- 8420227 TI - Elevated circulating levels of beta 2-microglobulin in patients with idiopathic dilated cardiomyopathy. PMID- 8420228 TI - Electrocardiographic features of atrial tachycardias after operation for congenital heart disease. PMID- 8420229 TI - Blood viscosity during thrombolytic therapy with anistreplase in acute myocardial infarction. AB - It has been postulated that a reduction in blood viscosity due to degradation of plasma fibrinogen may be of benefit to patients with acute myocardial infarction (AMI), who are treated with thrombolytic agents. The aims of this study were to investigate the time course of rheologic parameters, to identify the principal factors determining blood viscosity, and to find possible correlations between viscosity and cardiac function during thrombolytic therapy with anistreplase. Therefore, the viscosity of whole blood and plasma and the hematocrit were measured before and at 10 time points after thrombolysis in 10 patients with AMI. In addition, plasma fibrinogen and fibrin(ogen) degradation products were determined. Immediately after the start of thrombolysis, the viscosity of blood (both at high and low shear rate) and plasma decreased significantly and continued to do so for 24 hours. The mean hematocrit also decreased markedly, and even after correction for these hematocrit changes, the reduction in blood viscosity remained significant: it decreased to 72% of pretreatment (measured at low shear rate), whereas the high-shear viscosity decreased to 95% of baseline. The viscosity of plasma significantly decreased from 1.39 +/- 0.13 mPa.s (mean +/ SD) before thrombolysis to 1.22 +/- 0.08 mPa.s after 2 hours. There was a rapid, nearly complete depletion in fibrinogen, followed by a striking rebound after the second day. The decrease in blood viscosity lasted for 2 days after anistreplase and was mainly accounted for by the reduction in hematocrit. The contribution of fibrinogen to blood viscosity appeared less prominent. Despite these rheologic changes, no improvement in cardiac output was noticed in the patients. PMID- 8420230 TI - Effects of late (1 to 30 days) reperfusion after acute myocardial infarction on the signal-averaged electrocardiogram. AB - Early reperfusion (4 to 6 hours) after acute myocardial infarction reduces mortality and reduces the incidence of late potentials on a signal-averaged electrocardiogram (SAECG). Recent reports suggest that reperfusion accomplished after > 6 hours also may reduce mortality. The effect of such later reperfusion on the SAECG is not known. We hypothesized that reperfusion by angioplasty accomplished > 24 hours after onset of infarction would reduce late potentials and improve the parameters on the SAECG. Forty-one patients with a totally occluded infarct-related artery 12 +/- 8 days after infarction underwent attempted angioplasty. SAECG, echocardiography and thallium-201 imaging were performed before and 1 month after attempted angioplasty. Angioplasty resulted in successful reperfusion in 32 patients and was unsuccessful in 9. No change in the incidence of late potentials occurred after successful reperfusion (13 of 32 patients before and 13 of 32 patients 1 month later) or after unsuccessful reperfusion (6 of 9 patients before and 5 of 9 patients 1 month later). Among patients with successful reperfusion, no significant change occurred in the QRS duration or the terminal signal duration < 40 microV. The terminal root-mean square voltage in microvolts improved significantly at 1 month (31 +/- 25 before to 38 +/- 29 after, p = 0.004). Twenty-two of 32 patients with successful reperfusion had improved wall motion in the infarct zone at 1 month. Despite an improvement in function in these patients, no change in the incidence of late potentials occurred by 1 month.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420231 TI - Ventricular arrhythmias in rehabilitated and nonrehabilitated post-myocardial infarction patients with left ventricular dysfunction. AB - The incidence of ventricular arrhythmias in rehabilitated post-myocardial infarction (MI) patients with left ventricular dysfunction included in a long term rehabilitation program was assessed and compared with that in similar patients who were not in such a program. Thirty-eight post-MI patients (2 to 19 years after the acute event) with ejection fraction < 40% were investigated by 48 hour Holter monitoring. They were divided into the following 3 groups: group I, 11 patients who underwent arm training for 60 months; group II, 11 patients who underwent calisthenics for 36 months; and group III, 16 patients who were not in any rehabilitation program; the age of the patients was 61 +/- 7, 61 +/- 6 and 61 +/- 9 years, respectively, (p = not significant). Ejection fraction at rest was 31 +/- 9 for group I, 29 +/- 7 for group II, and 29 +/- 7 for group III (p = not significant). There were no significant differences concerning the location of MI, and antiarrhythmic treatment received by patients from all groups. At the conclusion of 48-hour Holter monitoring, 2 blood samples were obtained for assessment of norepinephrine (at rest and after postural change). Quality of life was determined by a detailed questionnaire, including questions concerning social activity, life satisfaction and sexual function. After 36 and 60 months, an improvement in hemodynamic condition of patients in group I was noted. Quality of life was higher in the rehabilitated patients, with enhanced emotional stability, satisfaction with work and social life, and a high percentage of return to work (82 vs 40%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420232 TI - Validation by high-frequency epicardial echocardiography of a new method of analyzing coronary angiography quantitatively in coronary artery disease. AB - In coronary atherosclerosis, the arterial lumen size and shape can be markedly irregular, eccentric and variable. Traditional angiographic interpretation, emphasizing percent diameter stenosis, has been criticized as an inadequate descriptor of such diseased arteries. Computerized quantitative angiographic technologies, yielding a true lumen area measurement, may be superior. High frequency epicardial echocardiography (HFEE) is a technique that allows on-line evaluation of coronary arterial wall and lumen at the time of cardiac surgery. It has been extensively validated and yields accurate measurements of normal and diseased coronary lumen areas. This study compares quantitative coronary angiography (QCA) estimates of lumen area to those obtained by HFEE to determine if the computerized angiographic method more accurately predicts residual luminal area than traditional angiographic percent diameter stenosis measurements. Although actual luminal morphology was quite variable, there was a good correlation between lumen areas determined by HFEE versus QCA: r = 0.85, n = 67, HFEE = 0.8 QCA - 0.1 (HFEE 4.0 +/- 0.30 mm2, mean +/- SEM range 0.3 to 14.0; QCA 5.1 +/- 0.40 mm2, range 0.7 to 11.8). Percent diameter stenosis determined from the angiograms did not correlate well with HFEE or QCA measurements of residual luminal area. Separation of "normal" arterial segments (defined as < 25% diameter stenosis) from "abnormal" segments (> 50% diameter stenosis) by angiography did not agree with lumen areas as defined by either HFEE or QCA. Better separation occurred when QCA-determined luminal areas were used to separate normal from abnormal arterial segments.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420233 TI - Prognostic value of dobutamine echocardiography in patients with high pretest likelihood of coronary artery disease. AB - To examine the value of transient regional asynergy on dobutamine stress echocardiography as a noninvasive predictor of future cardiac events, 51 symptomatic patients (aged 54 +/- 9 years) with suspected coronary artery disease (CAD) were studied using an incremental regimen of 5, 10, 15 and 20 micrograms/kg/min. Pretest likelihood of CAD was (mean +/- standard error of the percentage) 79.7 +/- 5.6% before and 83.4 +/- 5.2% after exercise electrocardiography using probability analysis based on age, sex and symptoms. Two-dimensional images were analyzed with reference to an 11-segment model and gave good interrater agreement. During 24 +/- 4 months (range 19 to 32) of follow up, 23 patients had events (1 myocardial infarction, 9 unstable angina, 10 coronary bypass surgery, 3 coronary angioplasty) and 28 were event free. Age, proportion with baseline asynergy and both pretest echocardiographic ejection fraction and its response to dobutamine were similar in these 2 groups (all p = not significant). Transient asynergy was seen in 17 of 23 patients (74%) with and 8 of 28 patients (29%) without events (p < 0.01); 5 of 6 patients (83%) with involvement of 3 segments had events. Myocardial infarction or unstable angina occurred in 8 of 25 (32%) with a positive and 2 of 26 (8%) with a negative stress echocardiogram (p < 0.05). Both exercise duration (389 +/- 195 vs 517 +/- 237 seconds, p < 0.05) and time to diagnostic ST-segment shift (291 +/- 192 vs 447 +/ 212 seconds, p = 0.05) were shorter in those with inducible asynergy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420234 TI - Prognostic value of noninvasive cardiac tests in the assessment of patients with peripheral vascular disease. AB - Two hundred thirty-six patients with peripheral vascular disease were prospectively studied to assess whether noninvasive cardiac investigations could predict prognosis better than simple clinical assessment. Clinical history, examination and resting electrocardiography were considered in all patients; exercise electrocardiography, Holter monitoring, radionuclide ventriculography and dipyridamole thallium imaging were performed in a subgroup of 168 patients. Follow-up for 6 to 30 months revealed major cardiac events in 21 patients. Cox survival analysis showed that clinical evidence of prior coronary artery disease was the best variable from clinical assessment that predicted cardiac events, with no other clinical variable adding to the statistical model. When variables from noninvasive cardiac assessment were added to the model, which included clinical evidence of coronary artery disease, dipyridamole thallium heart:lung ratio and left ventricular ejection fraction added significantly and incrementally to the prediction of cardiac events. Results of exercise electrocardiography or Holter monitoring did not add significantly. It is concluded that high lung uptake of thallium during dipyridamole stress, and impaired left ventricular ejection fraction help to identify patients with peripheral vascular disease who are at high cardiac risk, and should therefore be used for selecting subsequent cardiovascular medical, surgical and anesthetic management. PMID- 8420235 TI - Rationale and design of the Department of Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial (HIT) for secondary prevention of coronary artery disease in men with low high-density lipoprotein cholesterol and desirable low-density lipoprotein cholesterol. AB - Although a large body of epidemiologic evidence suggests that low levels of high density lipoprotein (HDL) cholesterol are strongly associated with an increased risk of coronary artery disease (CAD), no large-scale clinical trials focusing on this association have been reported. This report describes the rationale and design of the Department of Veterans Affairs HDL Intervention Trial (HIT), a multicenter, randomized, controlled clinical trial designed to determine whether lipid therapy reduces the combined incidence of CAD death and nonfatal myocardial infarction in men with established CAD who have low levels of HDL cholesterol with "desirable" levels of low-density lipoprotein (LDL) cholesterol. Twenty-five hundred men with CAD and HDL cholesterol < or = 40 mg/dl, LDL cholesterol < or = 140 mg/dl, and triglycerides < or = 300 mg/dl are being recruited at 20 Department of Veterans Affairs medical centers, randomized to either gemfibrozil or placebo, and followed in a double-blind manner for an average of 6 years. In this population, gemfibrozil is expected to increase HDL cholesterol by 10 to 15%, have a negligible effect on LDL cholesterol, and lower triglycerides by 30 to 40%. Because an estimated 20 to 30% of patients with CAD have a low HDL cholesterol as their primary lipid abnormality, the results of this trial are expected to have far-reaching clinical implications. PMID- 8420236 TI - Comparison of effects of propranolol versus pindolol on sinus rate and pacing frequency in sick sinus syndrome. AB - Beta blockers in patients with sick sinus syndrome (SSS) may prevent supraventricular arrhythmias, systemic hypertension and myocardial ischemia, but may cause excessive depression of sinus node function. In 8 patients with SSS and a permanent pacemaker, the effect of chronic oral pindolol on sinus rate and pacing frequency was compared with that of propranolol in a double-blind crossover trial. In all patients the pacemaker was programmed to a rate of < or = 50 beats/min. Holter monitors, obtained at baseline and on each drug, were used to calculate peak ambulatory sinus rate, number of paced beats per day, maximal number of paced beats per hour, and percentage of hours with paced beats. The peak sinus rate with pindolol therapy was 24% higher than with propranolol (p = 0.001). During pindolol therapy, the number of paced beats per day and maximal paced beats per hour were reduced 54% (p = 0.04) and 61% (p = 0.02), respectively, compared with propranolol. Patients with SSS who require beta blocker therapy for tachycardia, systemic hypertension or angina pectoris may have less bradycardia when treated with pindolol rather than propranolol. Beta blockers like pindolol, which cause less sinus node depression, may obviate the need for prophylactic permanent pacemakers in patients with SSS, and may help to prevent chronotropic incompetence and pacemaker syndrome in patients already treated with a VVI device. PMID- 8420237 TI - Delayed restoration of atrial function after conversion of atrial flutter by pacing or electrical cardioversion. AB - It is often suggested but never proven that atrial function is not affected during atrial flutter, nor after its conversion to normal sinus rhythm. To evaluate this hypothesis, a prospective study was performed in 22 patients (age range 20 to 88 years) with atrial flutter. Diastolic transmitral flow was analyzed with echo-Doppler before and after conversion. After randomization, conversion was attempted with overdrive pacing or up to two 50 J shocks. If the initial method was unsuccessful, a 200 J shock was administered. All patients were converted to sinus rhythm with this protocol. Shortly after conversion (at 1 and 6 hours), atrial contribution to ventricular filling was absent in 4 of 22 patients. In the remaining 18 patients, atrial contribution to ventricular filling was small. Atrial contribution to transmitral flow improved from 20 to 27% within 24 hours (p < 0.01) and increased further to 38% at 6 weeks (p < 0.005). Peak velocity of late diastolic filling increased from 0.28 m/s after 1 hour to 0.39 m/s after 24 hours (p < 0.0001) and improved even further during later follow-up. In 1 patient, an effective atrial systole was not observed until the 14th day. Cardiac output did not change significantly during the study period. No differences were observed between the conversion modalities. In conclusion, atrial dysfunction is present immediately after conversion of atrial flutter to normal sinus rhythm. This dysfunction occurs also after overdrive pacing and can last > 1 week. The findings suggest that stasis in the atria can remain temporarily present after successful conversion of atrial flutter to sinus rhythm. PMID- 8420238 TI - Use of multiple patches during implantation of epicardial defibrillator systems. AB - During implantation of epicardial automatic defibrillator systems, occasional patients have difficulty in obtaining adequate defibrillation thresholds. Of 236 consecutive patients undergoing implantation of epicardial defibrillator systems, 18 patients received a 3-patch (n = 15) or 4-patch (n = 3) defibrillator system. Twelve patients who received a multiple-patch defibrillator system had a best 2 patch defibrillation energy requirement of > or = 30 J; in the remaining 6 patients less stringent clinical criteria were used in the decision to add a third defibrillator patch (defibrillation energy requirement > 18 J in 4 patients, and > 20 J in 2 patients). Technically, multiple-patch systems were made possible with either the use of Y-connectors or defibrillators allowing output to 3 patches. In 3 patients, addition of a third epicardial patch still resulted in a defibrillation energy requirement of > or = 30 J; in these 3 patients, addition of a fourth patch resulted in a defibrillation energy requirement of < or = 20 J. All patients receiving a multiple-patch defibrillator system had a reduction in defibrillation energy requirement, and 12 patients had a reduction in defibrillation energy requirement of > or = 10 J over the best 2 patch defibrillation energy requirement. In the patients who eventually had placement of a multiple-patch system, the best 2-patch defibrillation energy requirement was > 18 J in 4 patients, > 20 J in 2 patients, > or = 30 J in 9 patients, and > 40 J in 3 patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420239 TI - Impaired immediate vasoconstrictor responses in patients with recurrent neurally mediated syncope. AB - Immediate responses to head-up tilt were determined in 78 consecutive patients with unexplained syncope undergoing 45-minute tilt tests at 60 degrees. Thirty four patients developed neurally mediated syncope (mean time to syncope 18 minutes), 40 tolerated the full duration of tilt, and 4 were unable to complete the study but did not develop syncope. Blood pressure, heart rate, forearm blood flow and forearm vascular resistance were measured at baseline and after 2 and 5 minutes of tilt. Syncopal and nonsyncopal patients were well-matched for age and baseline hemodynamic parameters. There was no difference between the groups in heart rate or blood pressure at 2 minutes, but there was a small but significant difference in percent reduction in mean arterial pressure at 5 minutes. After 2 and 5 minutes of tilt, mean forearm blood flow was 2.4 and 2.6 ml/min/100 ml, respectively, in syncopal patients compared with 1.6 (p < 0.05) and 1.7 ml/min/100 ml (p < 0.01), respectively, in patients who tolerated 45 minutes of tilt. In syncopal patients, forearm vascular resistance was 51.0 and 44.0 at 2 and 5 minutes, respectively, whereas in nonsyncopal patients, it was 82.4 (p < 0.02) and 73.1 (p < 0.001), respectively. These differences remained consistent when only data for patients developing syncope after > 15 minutes were included in the analysis. Patients with neurally mediated syncope have clearly demonstrable abnormalities in vascular control immediately after assumption of the upright posture. The results shed new light on the pathophysiology of neurally mediated syncope. PMID- 8420240 TI - A population-based estimate of candidacy rates for the implantable cardioverter defibrillator. AB - The implantable cardioverter-defibrillator (ICD) increases survival of patients who receive the device. However, candidacy rates have not been calculated for a defined population, and the potential effect of the device on the survival of all patients with heart disease has not been estimated. To make these calculations, medical records were reviewed for 1976 to 1988 in a population demographically similar to the white population of the United States. Definite and possible candidates were identified on the basis of American Heart Association/American College of Cardiology guidelines. Candidacy rates ranged from 3.3/100,000 (counting only definite candidates for the entire period) to 8.7/100,000 (counting definite and possible candidates after 1980). Extrapolated to the 1990 U.S. population, estimates ranged from 8,207 to 21,637 new candidates each year. During an average follow-up of 5 years, half of all deaths among candidates had the potential to be delayed by an ICD. In a similar population that has a death rate from heart disease of approximately 280/100,000, 0.6 to 1.6% of subjects have the potential to have their deaths delayed to some extent by an ICD. PMID- 8420241 TI - Comparison in acute myocardial infarction of anisoylated plasminogen streptokinase activator complex versus heparin evaluated by simultaneous thallium 201/technetium-99m pyrophosphate tomography. AB - In a subgroup of 45 patients with acute myocardial infarction (AMI) from the German multicenter trial of anisoylated plasminogen streptokinase activator complex (APSAC) (n = 20) versus heparin (n = 25), simultaneous thallium (TI)-201 technetium (Tc)-99m pyrophosphate (PYP) tomography was initiated to elucidate a possible benefit of APSAC over heparin. Findings in the 2 treatment groups were similar with respect to TI-201 defect score, relative scintigraphic infarct size, and in keeping with the main group coronary artery patency, global ejection fraction and maximal creatine kinase level. However, 2 different TI-201/Tc-99m PYP accumulation patterns within the area of infarction (homogeneous, group A; inhomogeneous, group B) were identified. Both treatment groups were similar with regard to the frequency of the homogeneous and inhomogeneous pattern. In comparing the 2 accumulation patterns, creatine kinase peaked earlier in group A than in group B, and global left ventricular ejection fraction was significantly higher in group A than in group B. In Group A, 30 of 31 patients and in group B 7 of 11 patients had a patent infarct-related vessel (p < 0.025). TI-201 defect score was lower in group A than in group B. Likewise, relative size of the infarction as determined from Tc-99m PYP images was significantly lower in group A than in group B. Fifteen patients experienced cardiogenic shock or severe heart failure. Patients in group B had a higher incidence of these in-hospital complications than patients in group A (92 vs 12%, p < 0.0005). Scintigraphic infarct size and TI-201 defect score were greater in patients with the aforementioned clinical events.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420242 TI - Effects of cardiac transplantation on endothelium-dependent dilation of the peripheral vasculature in congestive heart failure. AB - Patients with congestive heart failure demonstrate attenuated endothelium dependent vasodilation of the peripheral vasculature, but there are no data regarding the effect of therapies on this abnormality or whether this abnormality is reversible. This study was performed to address the hypothesis that abnormalities in endothelium-dependent vasodilation in heart failure are improved by heart transplantation. Forearm blood flow responses to the intraarterial administration of a dose range of methacholine, an endothelium-dependent vasodilator, and nitroprusside, an endothelium-independent vasodilator, were examined in 2 separate protocols. In protocol 1, forearm blood flow responses to methacholine in 14 heart transplant recipients were 5.02 +/- 3.11, 11.55 +/- 7.20 and 11.61 +/- 10.24 ml/min/100 ml forearm volume. These responses were significantly greater than those in 10 patients with heart failure (2.23 +/- 1.22, 4.60 +/- 3.43 and 6.70 +/- 4.91 ml/min/100 ml forearm volume; p < 0.05). In contrast, the responses to nitroprusside were nearly identical in the 2 groups. In protocol 2, six patients were studied before and 4 months (range 1 to 11) after transplantation. Methacholine responses before transplantation were 2.5 +/- 1.3, 5.2 +/- 1.5 and 7.3 +/- 1.5 ml/min/100 ml forearm volume and were significantly improved after transplantation to 5.7 +/- 1.2, 12.1 +/- 3.0 and 14.2 +/- 2.2 ml/min/100 ml forearm volume (p < 0.05). Peak reactive hyperemia responses increased significantly from 19.0 +/- 3.7 to 44.8 +/- 6.4 ml/min/100 ml forearm volume (p < 0.01) after transplantation. These data demonstrate that heart transplantation was associated with a significant improvement in the forearm blood flow responses to methacholine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420243 TI - Atrioventricular conduction in children of women with systemic lupus erythematosus. AB - The neonatal lupus syndrome consists of transient cutaneous lupus lesions or permanent congenital complete heart block (or hepatic fibrosis), or both, in infants born to mothers with systemic lupus erythematosus (SLE). The frequency of conduction abnormalities was examined in 86 offspring of 53 women affected by SLE. Electrocardiograms from the offspring demonstrated normal sinus rhythm in 84 of 86 offspring. The PR interval was normal for age (< 95th percentile) in 82 offspring and normal for heart rate in 81. Three children had a PR interval > 95th percentile (i.e., first-degree heart block) for both age and heart rate. The PR interval of the other 6 subjects with first-degree heart block for age or heart rate (> or = 95th percentile) was < or = 0.18 second. In contrast, using a rank assignment of PR intervals in relation to heart rate and age derived from published standards, grouped data indicated that heart rate adjusted for age was greater and PR interval adjusted for heart rate longer in offspring of mothers who had the onset of SLE before or during pregnancy than in the normal population; this observation did not hold for offspring whose mothers developed SLE after the pregnancy. These findings indicate that offspring of mothers with SLE, even in the absence of an abnormal electrocardiogram, may have experienced a maternal internal environment that produces subclinical changes in atrioventricular conduction. However, newborns with a normal pulse rate are unlikely to have significant abnormalities in atrioventricular conduction and do not need screening electrocardiograms at birth. PMID- 8420244 TI - Effect of exercise training on left ventricular performance in older women free of cardiopulmonary disease. AB - Endurance exercise training increases aerobic exercise capacity (maximal oxygen consumption rate [VO2max]) and attenuates the age-related decline in left ventricular (LV) function during exercise in older men. To determine whether similar adaptations occur in older women, 10 subjects (aged 63 +/- 4 years mean +/- SE) were studied before and after 9 to 12 months of endurance exercise training. They exercised 3.85 +/- 0.06 days/week at 81 +/- 0.3% of maximal heart rate. LV function at rest and during supine exercise was assessed by radionuclide ventriculography. VO2max was increased by 21% (from 1.40 +/- 0.1 to 1.7 +/- 0.1 liter/min; p < 0.001) in response to training. Maximal heart rate and systolic blood pressure during treadmill exercise were unchanged (161 +/- 5 beats/min before vs 164 +/- 3 beats/min after; p = NS, and 208 +/- 7 mm Hg before vs 214 +/ 8 mm Hg after; p = NS, respectively) after training. LV ejection fraction at rest (70.4 +/- 2% before vs 70 +/- 1% after) and during peak exercise (78.6 +/- 2% before vs 79.3 +/- 2% after) did not change in response to training. Furthermore, the increases in ejection fraction from rest to exercise were similar before and after training (change: 8.8 +/- 1 vs 9.1 +/- 1%). Stroke volume and cardiac output at peak exercise also did not change in response to training.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420245 TI - Chronic hepatitis: is it persistent, active, or just chronic? PMID- 8420246 TI - Crohn's disease presenting as isolated jejunitis years after childhood colitis. AB - We present a case of a patient who had documented ulcerative colitis as a child and later presented with isolated Crohn's jejunitis. Although rare, Crohn's disease must be considered in those patients with segmental inflammation of the small bowel and a prior history of inflammatory bowel disease involving the colon. Patients with colitis that do not show specific criteria for either ulcerative colitis or Crohn's disease should be classified as indeterminant colitis, and a small bowel series is indicated. It may be indicated to obtain a small bowel series in patients with any form of colonic inflammatory bowel disease, periodically, after diagnosis. PMID- 8420247 TI - Peritoneal dialysis therapy for patients with liver and renal failure with ascites. AB - Two patients with severe liver disease complicated with ascites were recently treated at our institution. Both rapidly developed renal failure. In one patient, liver disease was the result of alcohol abuse, and in the other, was due to malnutrition associated with obesity and acute weight loss. The only reasonable therapeutic approach for these patients was believed to be a course of peritoneal dialysis, along with other supportive measures. In both cases, the management was successful. Furthermore, it was possible to discontinue dialysis at the time of discharge. We conclude that peritoneal dialysis can be a life-saving procedure in patients with severe liver disease and ascites complicated by renal failure. PMID- 8420248 TI - The relationship between stress and symptoms of gastroesophageal reflux: the influence of psychological factors. AB - This paper describes the first controlled study of the relationships among stress, psychological traits associated with chronic anxiety, acid reflux parameters, and perceptions of reflux symptoms. Seventeen subjects with symptomatic reflux disease were studied using a 2 (high vs. low gastrointestinal susceptibility score) x 2 (stress vs. neutral tasks) x 3 (periods 1, 2, or 3) experimental design. It was found that the stress tasks produced significant increases in systolic and diastolic blood pressure, pulse rates, and subjective ratings of anxiety and reflux symptoms. The stress tasks, however, did not influence objective parameters of acid reflux (total acid exposure, number of reflux episodes, duration of longest reflux episode). Moreover, the effect of stress on reflux ratings was due primarily to the responses of the subjects with high gastrointestinal susceptibility scale scores. These subjects' reflux ratings remained at high levels during all stress periods, whereas subjects in all other experimental conditions reported decreased reflux symptoms across periods. These results suggest that reflux patients who are chronically anxious and exposed to prolonged stress may perceive low intensity esophageal stimuli as painful reflux symptoms. Future effort should be devoted to examining the efficacy of anxiolytic and behavioral therapies with these reflux patients. PMID- 8420249 TI - Magnetic resonance imaging appearance of hepatic schistosomiasis, with ultrasound and computed tomography correlation. PMID- 8420250 TI - Pancreatic duct obstruction as a potential etiology of pancreatic adenocarcinoma: a clue from pancreas divisum. PMID- 8420251 TI - Fistulas are not always due to Crohn's disease: an unusual groin abscess. AB - A 38-yr-old white female with Crohn's disease was admitted for right hemicolectomy to treat an enterocutaneous fistula of 4 yr duration. At laparotomy, it was found that the fistula was related to an appendiceal diverticulum, rather than Crohn's disease, and simple appendicectomy was curative. The literature is reviewed. PMID- 8420252 TI - Fulminant acute colitis following a self-administered hydrofluoric acid enema. AB - A 33-yr-old white male presented with bloody diarrhea, leukocytosis, and left lower quadrant direct and rebound tenderness after a self-administered concentrated hydrofluoric acid enema while intoxicated from intranasal cocaine administration. Intraoperative flexible sigmoidoscopy and a gastrografin enema revealed severe mucosal ulceration and edema in the rectum and sigmoid colon. Laparotomy revealed an ulcerated, necrotic, and purulent sigmoid colon and intraperitoneal pus. The patient underwent a limited sigmoid resection and a Hartman procedure. Five months later, the patient presented with a rectal stricture which was resected. This case demonstrates that a hydrofluoric acid enema can cause fulminant acute colitis and chronic colonic strictures. PMID- 8420253 TI - Emphysematous gastritis secondary to ingestion of large amounts of Coca Cola. AB - Acute emphysematous gastritis (AEG) is a life-threatening disease in which gas forming bacteria invade the gastric wall and cause acute inflammation of it. The clinical presentation of the patient with AEG is stormy: severe sepsis which usually leads to an early death. Presented herein is a case of a 16-yr-old boy with AEG. There were no predisposing factors to the condition in this case. Gas invaded the stomach wall and portal venous system, as well as the duodenal wall, a finding that has not been reported previously. The clinical course was very severe but, in contrast to previously reported cases, recovery was very rapid and left no sequelae. PMID- 8420254 TI - Endometriosis of the small intestine presenting as a protein-losing enteropathy. AB - A 46-yr-old multiparous cachetic woman presented with severe hypoalbuminemia in the absence of liver disease, proteinuria, and/or protracted starvation. The clinical presentation and work-up was indicative of protein-losing enteropathy. She developed an acute partial small bowel obstruction, and a presumptive diagnosis of lymphoma of the small intestine was entertained. Surgical resection of the terminal ileum revealed transmural involvement of the bowel by endometriosis. Her postoperative recovery was uneventful, with return of her serum albumin levels to normal. PMID- 8420255 TI - Mesenteric venous thrombosis in familial free protein S deficiency. AB - Protein S deficiency reportedly is associated with a high risk for thromboembolism, but mesenteric venous thrombosis is a rare manifestation of this hereditary disorder. We describe an additional case of mesenteric venous thrombosis in a 25-yr-old man with type I protein S deficiency. Total protein S level remained just below the normal range, but free protein S level was less than 10% of normal. Family study revealed that his father had had the same type of disorder. We have reviewed six well-documented cases of mesenteric venous thrombosis with protein S deficiency. It is particularly interesting that those with type I deficiency have had no previous thrombotic episodes (p < 0.05). Our patient was successfully treated with extended bowel resection, but unusual perioperative management was required. Because of extremely edematous mesentery and small bowel, we had to treat him with the abdomen wide open for 2 days between the primary and second laparotomies. PMID- 8420256 TI - Giant colonic diverticulum: an unusual manifestation of a common disease. AB - Giant colonic diverticula are an uncommon manifestation of colonic diverticular disease. This report documents another case of this rare entity. The clinical presentation is variable, but abdominal pain and the presence of an abdominal mass are common. Their etiology is uncertain, but they are thought to originate from pulsion diverticula. Abdominal plain films and barium enema are helpful in making the diagnosis. Computerized tomography is useful when the diagnosis is unclear. Surgical resection is recommended to alleviate symptoms and avoid complications. PMID- 8420257 TI - Long-term survival of a patient with bile duct carcinoma by endoscopic biliary drainage. AB - An 80-yr-old female presented with obstructive jaundice. Endoscopic retrograde cholangiopancreatography showed a carcinoma in the middle extrahepatic bile duct, and a biliary endoprosthesis was inserted. Exfoliative cytology of the bile and forceps biopsy of the tumor revealed a papillary adenocarcinoma. Surgical resection could not be done because of her cardiovascular complications, and neither chemotherapy nor radiotherapy was administered. Stents were exchanged and cleaned 21 times because of occlusion and cholangitis. Subsequent serial cholangiogram showed a slow growth of the papillary tumor, but local invasion to the adjacent organs or distant metastasis was not observed. The patient survived for 7 yr and 6 months after insertion of the biliary endoprosthesis. PMID- 8420258 TI - Mucin as a marker for aggressiveness of colon cancer. PMID- 8420259 TI - Screening for curable colorectal tumors: an approach for the 21st century. PMID- 8420260 TI - Omeprazole dosing: AM vs. PM: does sunset make a difference? PMID- 8420261 TI - Heater probe dislodgement during use. PMID- 8420262 TI - Use of glycyrrhizin for recurrence of hepatitis B after liver transplantation. PMID- 8420263 TI - Psyllium worm. PMID- 8420264 TI - More about omeprazole-cyclosporine interaction. PMID- 8420265 TI - Probable hepatorenal syndrome at presentation. PMID- 8420266 TI - Cholelithiasis in cirrhotic patients from Turkey. PMID- 8420267 TI - Pancreatic carcinoma associated with situs inversus in a young adult. PMID- 8420268 TI - Interaction between patient and test administrator may influence the results of edrophonium provocative testing in patients with noncardiac chest pain. AB - Edrophonium is a widely used provocative agent in the evaluation of noncardiac chest pain, with reported positivity rates of 30-55%. The influence of a subjective response and psychological factors on test results have not been examined previously. A retrospective analysis was performed to compare positivity rates for three physicians in the same laboratory. This was followed by a prospective study of 62 patients with noncardiac chest pain randomized to two groups. Group 1 patients were told that intravenous medication was given to observe changes in the tracing. Group 2 patients were told that the injection was to elicit their usual pain. During the 2-yr retrospective review, 260 patients were tested. The positivity rate varied from 31.1% with physician A to 20.2% with physician B and 7.5% for physician C (p = 0.001 for A vs. C, and p = 0.04 for B vs. C). In the prospective study, chest pain was elicited in nine of 62 patients (14.5%). Two of the 29 patients in group 1 (6.9%) and seven of 33 patients in group 2 (21.2%) contributed to this result. Contraction amplitude and duration increased similarly in all groups. These data suggest that edrophonium testing may be influenced by coaching, that manometric changes are similar in positive and negative tests, and that the prevalence of positive tests is lower than previously reported. PMID- 8420269 TI - Normal and abnormal proximal esophageal acid exposure: results of ambulatory dual probe pH monitoring. AB - Ambulatory esophageal pH monitoring was performed in 26 normal volunteers, 20 patients with normal distal acid exposure, and 23 patients with abnormal distal acid exposure in an attempt to define normal values for proximal esophageal acid exposure using a standardized technique. We used a dual pH sensor with antimony electrodes spaced at 15 cm. The distal electrode was placed manometrically at 5 cm above the lower esophageal sphincter. Proximal electrode thus was located below the upper esophageal sphincter in the esophageal inlet. The patients underwent 24-h ambulatory pH monitoring and were told to pursue normal daily activities. The percentage of acid exposure time and number of episodes per 24 h at both pH < 4.0 and 5.0 were measured for the total, upright, and supine periods. Since the pH values were not normally distributed, the medians and 95th percentiles were used to define normal values. Minimal acid exposure occurred in the proximal esophagus (< 1% total; 0% supine) in volunteers and patients with normal distal reflux. Patients with abnormal distal acid exposure had significantly greater proximal reflux. PMID- 8420270 TI - Investigation of gastrointestinal bleeding in patients with end stage renal disease. AB - In order to evaluate the source and course of gastrointestinal bleeding in patients with established renal insufficiency, we reviewed data on 40 patients with renal failure and gastrointestinal bleeding seen over 2 yr. A randomly selected control group of 39 patients without renal failure was used for comparison. Medical records of our University Hospital were reviewed, and patients with a documented gastrointestinal bleed and renal insufficiency (creatinine greater than 1.7 mg/dl) were included in this study. Panendoscopy was the most valuable procedure in terms of establishing a diagnosis as to the cause of bleeding. Colonoscopy was of questionable value unless the bleed was clearly of lower intestinal origin. Recurrent bleeding during the index admission occurred with the same frequency in both groups of patients. Both groups used nonsteroidal anti-inflammatory agents frequently. The findings and outcome for this group of patients with renal failure was comparable to the control patients with gastrointestinal bleeding. PMID- 8420271 TI - Graded pneumatic dilation using Rigiflex achalasia dilators in patients with primary esophageal achalasia. AB - Pneumatic dilation is the initial therapy for primary esophageal achalasia. Recently, polyethylene balloon (Rigiflex) dilators have been used with varying success and complication rate. We performed a total of 47 dilations in 29 consecutive patients with achalasia using the Rigiflex dilators. The 3.0-cm balloon was always used first. If there was no symptomatic response, a 3.5-cm balloon was used after 4-8 wk. If there was still no symptomatic response after 4 8 wk, a 4.0-cm dilator was used. Eighteen (62%) patients were successfully dilated with a 3.0-cm balloon only. Of 11 patients not responding to a 3.0-cm balloon, five were dilated successfully with a 3.5-cm balloon. Of six patients not responding to a 3.5-cm balloon, four were successfully dilated with a 4.0-cm balloon dilator. Two patients eventually required surgery. The overall success with Rigiflex balloon dilator was achieved in 27 of 29 (93%) patients. There were no complications. We conclude that pneumatic dilation for esophageal achalasia performed in a graded fashion starting with Rigiflex 3.0-cm balloon dilator has a high success rate without complications in patients with achalasia. PMID- 8420272 TI - Immunosuppressive therapy in pediatric inflammatory bowel disease: results of a survey of the North American Society for Pediatric Gastroenterology and Nutrition. Subcommittee on Immunosuppressive Use of the Pediatric IBD Collaborative Research Forum. AB - We report the results of a survey of the membership of the North American Society for Pediatric Gastroenterology and Nutrition designed to determine pediatric gastroenterologists' attitudes toward the use of immunosuppressive therapy for inflammatory bowel disease (IBD), and to assess how these medications are actually being used in the treatment of children with IBD. One hundred five physicians (27% of surveys) responded. Eighty-eight (84%) had prescribed 6 mercaptopurine and/or azathioprine for IBD, and 66 believed that they were effective. Only 12 had used cyclosporine and four methotrexate. All physicians who had used immunosuppressives in IBD had prescribed them for patients with Crohn's disease, but only 50% had prescribed them for ulcerative colitis. The predominant indications for use included intractable symptoms despite traditional medical therapy (92%) and for corticosteroid-sparing effects (86%). Potential toxicities of greatest concern included marrow and immune suppression and malignancy. The vast majority of responders were not certain what to recommend with respect to the use of immunosuppressive agents prior to and during pregnancy. A clinical database was compiled from 165 retrospective case reports submitted by 45 physicians (33 medical facilities). At the start of immunosuppressive therapy, patients were 15.3 +/- 4.0 yr of age, and 52% were Tanner IV-V. Eighty-one percent had Crohn's disease, 8% ulcerative colitis, and 11% indeterminant colitis. One hundred twenty-two were treated with 6 mercaptopurine, and 43 with azathioprine. Five also received cyclosporine concomitantly. Overall, 68% of patients treated with an immunosuppressive improved. Complications requiring discontinuation of immunosuppressive therapy occurred in 6% of patients. It appears that immunosuppressives are commonly used to treat children with IBD despite a paucity of data regarding their safety and efficacy in this age group. Controlled, prospective trials are warranted to better define the role of immunosuppressive therapy in pediatric IBD. PMID- 8420273 TI - Colonic lavage solutions: plain versus flavored. AB - Many patients dislike the taste of the oral gastrointestinal lavage solutions utilized prior to colonoscopy. Specifically, patients dislike the salty taste and the quantity of the polyethylene glycol-electrolyte solution that they are required to ingest. In an attempt to reduce the salty taste and potentially improve patient compliance and tolerance of the preparation, flavoring was added to the lavage solution. Fifty-six patients received standard or flavored polyethylene glycol-electrolyte lavage solution in random order prior to colonoscopy. In group I, lemon-flavored Colyte was compared with standard Colyte: 93% (25/27) of subjects preferred the flavored solution. In group II, standard Colyte was compared with lemon/lime-flavored Colyte, and 80% (8/10) of subjects preferred standard Colyte. In group III, lemon-flavored Colyte, standard Colyte, and NuLYTELY were compared. Ninety percent (17/19) of the subjects preferred lemon-flavored Colyte, 10% of the subjects preferred standard Colyte, and none of the subjects preferred NuLYTELY. Overall, patients preferred lemon-flavored solution over other solutions. PMID- 8420274 TI - Enteric protein loss during shigellosis. AB - To determine whether protein-losing enteropathy occurs during shigellosis, we measured concentrations of alpha 1-antitrypsin in sequential stool samples from 110 adults hospitalized with acute dysentery due to Shigella infection. Mean stool concentrations of alpha 1-antitrypsin on admission were 10.9 micrograms/mg dry weight of stool. Stool alpha 1-antitrypsin concentrations were significantly (p < 0.001) lower on the 3rd and 5th study days (4.1 and 2.2 micrograms/mg, respectively) than on admission. Admission mean alpha 1-antitrypsin concentrations in patients with Shigella dysenteriae type 1 infection (14.4 micrograms/mg) were significantly (p < 0.05) higher than in patients infected with other species of Shigella (9.3 micrograms/mg). Stool alpha 1-antitrypsin concentrations were significantly correlated with the number of erythrocytes in the stool, and inversely correlated with serum protein concentration. Patients in whom antimicrobial treatment failed, most often because they were infected with a resistant strain of Shigella, had significantly higher concentrations of alpha 1 antitrypsin on all three study days. We conclude that Shigella infection is associated with a protein-losing enteropathy, that this enteropathy is more severe with S. dysenteriae type 1 infection, and that the enteropathy improves with appropriate antimicrobial therapy. PMID- 8420275 TI - A clinical comparison of an electrohydraulic and a piezoelectric shockwave lithotripter in gallstone therapy. AB - The aim of this prospective, randomized study was to compare two second generation lithotripters based on different physical principles in patients with gallbladder stones at a single lithotripsy center under the same clinical conditions. Sixty patients with one to three symptomatic gallbladder stones were selected for lithotripsy, either with an electrohydraulic or a piezoelectric device. With both lithotripters, treatment was performed under standard conditions (prone position, sonographic monitoring, sedoanalgesia if necessary, up to 3000 pulses/session, retreatments (maximum, two) if fragments > 4 mm, concomitant oral chemolitholysis). If no fragmentation could be obtained in the first session, the other lithotripter was used for the following treatments. The two groups did not differ significantly with regard to the anthropometric data or number and size of stones. In contrast to piezoelectric lithotripsy (0%), with the electrohydraulic lithotripter, iv analgesics and sedatives were necessary in all treatments (100%); however, in 11/53 treatments (21%), patients did not tolerate the full session despite maximum medication. The treatment time was nearly twice as long with electrohydraulic (56 +/- 22 min) than with piezoelectric lithotripsy (31 +/- 8 min) (p < 0.001). With the electrohydraulic lithotripter, used in 20% of the patients, no fragmentation was seen after the first session, and therapeutically adequate fragmentation (< or = 4 mm) occurred in only 33%. In contrast, with the piezoelectric lithotripter, the stones were disintegrated in all patients (p < 0.05); in 50% a maximum fragment size < or = 4 mm was measured after the first treatment. Whereas in the first months after lithotripsy, stone-free rates were higher with piezoelectric lithotripsy (43% vs. 25% after 1 month; 47% vs. 38% after 2 months; 60% vs. 48% after 4 months; NS), rates of complete stone disappearance were equally high in both groups after 12 months (82%). PMID- 8420276 TI - Eosinophilic gastroenteritis: 10 years experience. AB - Eight patients (five men, three women), mean age 36.9 +/- 13.5 (17-60) yr, were diagnosed to have eosinophilic gastroenteritis. The condition was proved in five patients by biopsies through endoscope, and in three, by operation. All had hypereosinophilia (absolute eosinophil count of 1,337-21,787/cm3). According to Klein classification, two had mucosal disease, three had muscle layer disease, and three, subserosal disease. The most common symptoms were abdominal pain (100%), diarrhea (62.5%), vomiting (62.5%), and nausea (50%). Four patients (50%) had symptoms for more than 1 yr before diagnosis. Barium studies revealed mucosal edema and/or thickening of the small intestinal wall in three cases (including one case with ulceration), partial gastric outlet obstruction in one case, and narrowing of lumen of the terminal ileum in one case. Hypotonic duodenogram revealed double contour in one case. Ultrasound examination revealed thickening of the intestinal wall in two cases; computer tomography revealed thickening of the intestinal wall in one case. All patients were treated with steroid (40 mg/day for initial dose and relapse; 5-10 mg/day for maintenance). The symptoms subsided and the eosinophil counts returned to normal within 2 wk in seven patients and 1 month in one. Of six patients being followed up for 2-10 yr, one had remission, four needed small maintenance dose of steroid, and one suffered from relapse with intestinal perforation. PMID- 8420277 TI - Abdominal tuberculosis in Saudi Arabia: a clinicopathological study of 65 cases. AB - The clinical and pathological features of 65 patients with abdominal tuberculosis obtained during a 7-yr period were analyzed and the diagnostic procedures critically evaluated. The diagnosis was histologically confirmed in 59 patients. In two more patients, the diagnosis was based solely on a positive ascitic fluid culture for tubercle bacilli. The remaining four patients responded dramatically to anti-tuberculous chemotherapy given on suspected laparoscopic findings in cases in which no biopsy was taken. Laparoscopy was found to be safer and superior to laparatomy and is recommended as an initial investigation in the diagnostic work-up of patients in whom tuberculous peritonitis is suspected. Furthermore, the finding of granulomatous inflammation in peritoneal biopsy is a justification for immediate therapy in such patients. This is particularly valid in endemic areas if one considers the risks of delaying treatment of these patients. PMID- 8420278 TI - Studies on the relationship between hepatic and splenic density change in dynamic CT in the presence of a large splenorenal shunt in patients with liver cirrhosis. AB - Using dynamic computerized tomography (CT), we studied liver and spleen hemodynamics in one group of patients with a large splenorenal shunt and another group without a large shunt. Specifically, we measured the time-to-peak hepatic and splenic enhancement and the loss of enhancement, i.e., decay time. The time to-peak liver enhancement (61 +/- 21 s vs. 81 +/- 14 s, p < 0.05) and the splenic decay time (24 +/- 16 s vs. 48 +/- 28 s, p < 0.05) were significantly shorter in patients with a large splenorenal shunt compared to those without. The groups had similar hepatic decay times and times to peak splenic enhancement. We found dynamic CT useful in assessing the effect of a large splenorenal shunt on portal and splenic hemodynamics. PMID- 8420279 TI - Role of endogenous prostaglandins in secretin- and plaunotol-induced inhibition of gastric acid secretion in the rat. AB - We investigated the role of endogenous prostaglandins in the inhibitory effect of exogenous secretin and the antiulcer agent plaunotol on gastric acid secretion in the rat. Intravenous infusion of secretin (0.05 CU/kg/h) and intraduodenal administration of the secretin-releasing agent, plaunotol (320 mg/h), resulted in significant inhibition of gastric acid secretion stimulated by intravenous infusion of pentagastrin (0.3 micrograms/kg/h), and this was accompanied by an increase in the prostaglandin E2 content of the gastric mucosa. Intraduodenal administration of plaunotol (320 mg/h) produced plasma secretin levels comparable to the levels achieved by intravenous infusion of secretin (0.05 CU/kg/h). Intravenous administration of prostaglandin synthesis inhibitor, indomethacin (2 mg/kg + 1 mg/kg/h), completely abolished both the inhibitory action of secretin and plaunotol on gastric acid secretion, and the increase in gastric mucosa prostaglandin E2 induced by secretin and plaunotol. The results indicate that endogenous prostaglandins play a significant role in the inhibitory action of exogenous and plaunotol-released endogenous secretin in the rat. PMID- 8420280 TI - Successful endoscopic injection sclerotherapy of a bleeding duodenal varix. AB - Bleeding from duodenal varices is an unusual event. We report the case of a 50-yr old man with portal hypertension due to alcoholic cirrhosis who presented with upper gastrointestinal bleeding and encephalopathy. Emergent endoscopy revealed an actively bleeding duodenal varix. The bleeding was treated successfully with injection sclerotherapy. Only four cases of injection sclerotherapy of bleeding duodenal varices have been reported previously. We review and compare reported cases of sclerotherapy of duodenal varices and also review the other therapeutic options. Endoscopic injection sclerotherapy of bleeding duodenal varices appears to be a useful first-line therapy. PMID- 8420281 TI - Disappearance of duodenal polyps in Gardner's syndrome with sulindac therapy. AB - Duodenal polyps with a malignant potential pose a serious threat to Gardner's patients. Several reports have shown regression or disappearance of colonic polyps with sulindac therapy. We report the first case of disappearance of duodenal polyps with sulindac. PMID- 8420282 TI - Octreotide reduces enteral protein losses in Menetrier's disease. AB - A 47-yr-old man with Menetrier's disease presented with profound enteral protein losses documented by 51Cr-albumin stool studies. There was no response to anticholinergics, H2-receptor antagonists, or proton pump inhibition. Subcutaneous octreotide acetate, 100 micrograms bid, resulted in clinical improvement with rapid resolution of enteral protein losses. Octreotide was continued for 12 months. During an additional 5-month follow-up period without therapy, the patient remained improved clinically with low to normal serum proteins, despite the persistence of gastric rugal hypertrophy. The evidence that octreotide resolved enteral protein losses and eliminated the need for urgent gastrectomy in our patient warrants a trial of this drug for other patients with Menetrier's disease. PMID- 8420283 TI - Chyloperitoneum associated with chronic severe sarcoidosis. AB - A 51-year-old black female with sarcoidosis diagnosed 20 yr earlier developed massive chyloperitoneum and chylothorax. Autopsy demonstrated fibrotic and enlarged thoracic and mesenteric lymph nodes. Sarcoidosis can produce chyloperitoneum by producing intrathoracic nodal fibrosis and lymphatic obstruction. PMID- 8420284 TI - Low high-density lipoprotein cholesterol: what does it mean, what can we do about it, and what should we do about it? PMID- 8420285 TI - Candida prosthetic arthritis: report of a case treated with fluconazole and review of the literature. AB - Prosthetic arthritis due to Candida species is uncommon, with only 15 cases reported in the literature. We recently cared for a human immunodeficiency virus infected patient who developed Candida parapsilosis prosthetic arthritis unresponsive to resection arthroplasty, intravenous amphotericin B, and suppressive ketoconazole therapy. Treatment with fluconazole led to mycologic cure and symptom improvement, although he subsequently underwent above-the-knee amputation due to continued joint instability. Fluconazole may be useful follow up therapy after a course of amphotericin B combined with resection arthroplasty or when removal of the prosthesis cannot be accomplished. PMID- 8420286 TI - Cutaneous leukocytoclastic vasculitis and renal cancer: two cases. AB - We report two cases of cutaneous leukocytoclastic vasculitis (CLV) leading to the discovery of an as yet asymptomatic, surgically curable clear cell carcinoma of the kidney. CLV causative factors or associated diseases are usually drugs, infection, or collagen vascular disease, but rarely malignancies. In such cases, these are more often malignant hematologic diseases than solid neoplasms. We believe that in apparently idiopathic CLV, a screening examination should be done to detect any underlying early-stage curable solid malignancy. PMID- 8420287 TI - Simvastatin-induced rhabdomyolysis followed by a MELAS syndrome. PMID- 8420288 TI - Polymyositis associated with anti-Jo1 antibodies: severe cardiac involvement as initial manifestation. PMID- 8420289 TI - Home antibiotic therapy for streptococcal endocarditis: a call for a controlled trial. PMID- 8420290 TI - Elevated creatine phosphokinase and creatine phosphokinase-MB in hyperventilation. PMID- 8420291 TI - Home intravenous antibiotic therapy. PMID- 8420292 TI - Home intravenous antibiotic therapy. PMID- 8420293 TI - Cruzan II. PMID- 8420294 TI - Evaluation of ethics consultations. PMID- 8420295 TI - Evaluative models of ethics consultation. PMID- 8420296 TI - Pravastatin and gemfibrozil alone and in combination for the treatment of hypercholesterolemia. AB - PURPOSE: To compare the efficacy and safety of pravastatin, gemfibrozil, combined therapy, and placebo in the treatment of hypercholesterolemia. PATIENTS AND METHODS: At 5 centers in Sweden and 2 in Finland, 290 ambulatory patients were randomized to active treatment or placebo for 12 weeks following a single-blind placebo lead-in period. The study was double-blind and placebo-controlled. Patients has plasma total cholesterol levels of at least 6.0 mmol/L or in the 90th percentile by age and sex and triglycerides less than 4.0 mmol/L. Concentrations of lipids, lipoproteins, and apolipoproteins were measured, and clinical laboratory tests included liver function and creatine kinase determinations. RESULTS: Pravastatin reduced total cholesterol (26.3% versus 15.2%, p < or = 0.01), low-density lipoprotein cholesterol (LDL-C) (33.5% versus 16.8%, p < or = 0.01), and apolipoprotein B (28.8% versus 15.3%, p < or = 0.01) more than gemfibrozil. Gemfibrozil reduced very-low-density lipoprotein cholesterol (VLDL-C) (49.1% versus 21.9%, p < or = 0.01) and triglycerides (42.2% versus 14.2%, p < or = 0.01) and increased high-density lipoprotein cholesterol (HDL-C) (15.2% versus 5.9%, p < or = 0.01) more than pravastatin. Pravastatin and gemfibrozil increased apolipoprotein A-I comparably (3.3% versus 5.0%, p = NS). The combination significantly (p < or = 0.01) reduced total cholesterol (29.0%), LDL-C (37.1%), VLDL-C (49.4%), and apolipoprotein B (31.6%), and increased HDL-C (16.8%). The combination reduced the total cholesterol/HDL-C (39.3%) and LDL C/HDL-C (45.8%) ratios significantly (p < 0.01). Adverse events and clinical laboratory abnormalities were generally mild and transient in all groups, although creatine kinase tended to be higher with combination therapy. Study drugs were withdrawn from two patients with asymptomatic creatine kinase elevations. Severe myopathy was not observed; however, the presence of subclinical musculoskeletal effects cannot be excluded. CONCLUSIONS: Co administration of pravastatin and gemfibrozil combined the specific effects of the two drugs on lipoprotein concentrations and ratios. The incidence of side effects was low; severe myopathy did not occur. The combination may be useful in selected cases of combined hyperlipidemia; however, since myopathy at a low incidence or after long-term therapy cannot be excluded, the routine use of combination therapy is not advisable. PMID- 8420297 TI - Anaerobic osteomyelitis and arthritis in a military hospital: a 10-year experience. AB - PURPOSE: The methods of collecting, transporting, cultivating, and identifying aerobic bacteria in bone and joint infections have improved markedly since the early 1980s. In addition, many of the anaerobes have been reclassified and renamed. The purpose of this study was to provide more current information regarding the incidence of recovery of anaerobic bacteria from clinical specimens of infected bone and joint. MATERIALS AND METHODS: Specimens from 73 infected bone specimens and 65 infected joints inoculated on media supportive for aerobic and anaerobic bacteria showed bacterial growth. RESULTS: One hundred fifty-seven organisms (2.2 isolates/specimen), consisting of 122 anaerobic bacteria (1.7 isolates/specimen) and 35 facultative or aerobic bacteria (0.5 isolate/specimen), were recovered from the 73 bone specimens. Anaerobic bacteria were recovered with aerobe or facultative bacteria in 24 (33%) instances. The predominant anaerobes were Bacteroides species (49 isolates), anaerobic cocci (45), Fusobacterium species (11), Propionibacterium acnes (7), and Clostridium species (6). Conditions predisposing to bone infections were vascular disease, bites, contiguous infection, peripheral neuropathy, hematogenous spread, and trauma. Pigmented Prevotella and Porphyromonas species were mostly isolated in skull and bite infections (7 of 19), members of the Bacteroides fragilis group in hand and feet infection (12 of 16), and Fusobacterium species in skull, bite, and hematogenous long bone infections. Seventy-four organisms (1.1 isolates/specimen), consisting of 67 anaerobic bacteria (1.0 isolate/specimen) and 7 facultative or aerobic bacteria (0.1 isolate/specimen), were isolated from 65 joint specimens. The predominant anaerobes were P. acnes (24 isolates), anaerobic cocci (17), Bacteroides species (10), and Clostridium species (5). Predisposing conditions to joint infection were trauma, prior surgery, presence of a prosthetic joint, and contiguous infection. P. acnes isolates were associated with prosthetic joints, members of the B. fragilis group with hematogenous spread, and Clostridium species with trauma. The clinical presentation of these cases is discussed. CONCLUSION: These data highlight the importance of anaerobic bacteria in bone and joint infection. PMID- 8420298 TI - Long-term results of monthly inhaled pentamidine as primary prophylaxis of Pneumocystis carinii pneumonia in HIV-infected patients. AB - PURPOSE: To evaluate the long-term efficacy and safety of inhaled pentamidine as primary prophylaxis against Pneumocystis carinii pneumonia (PCP) in patients infected with human immunodeficiency virus (HIV). PATIENTS: Two hundred thirty two HIV-infected patients with a CD4 cell count below 20% of the total lymphocyte count were given aerosolized pentamidine once every 4 weeks for more than 3 months. Pentamidine aerosols were administered at the hospital under medical supervision. Prevention of bronchospasm was carried out using inhaled salbutamol. RESULTS: Mean duration of prophylaxis was 15.9 months. Eleven patients (4.7%; [95% confidence interval 2% to 7.4%]) developed PCP. Probability to remain free of PCP is 95.6% at 12 months, 94% at 18 months, and 88% at 24 months. Mean delay between the onset of the prophylaxis and the occurrence of PCP for the 11 patients was 12.9 months (range: 4 to 26 months). No major side effect was observed, and minor side effects (cough, acute dyspnea) were infrequent. CONCLUSION: The efficacy and tolerance of aerosolized pentamidine as shown in our study support its use as primary prophylaxis against P. carinii in HIV-infected patients. PMID- 8420299 TI - Interrelation of humoral factors, hemodynamics, and fluid and salt metabolism in congestive heart failure: effects of extracorporeal ultrafiltration. AB - PURPOSE: We investigated the mechanisms involved in the regulation of salt and water metabolism in patients with congestive heart failure (CHF). Extracorporeal ultrafiltration was utilized as a nonpharmacologic method for withdrawal of body fluid. PATIENTS, METHODS, AND RESULTS: In 32 consecutive patients with CHF (New York Heart Association functional class II to IV) and different degrees of water retention, 24-hour diuresis and natriuresis were inversely best correlated with the combination of circulating renin, aldosterone, norepinephrine, and renal perfusion pressure (RPP, mean aortic pressure minus mean right atrial pressure). Fluid withdrawal (600 to 5,000 mL) at a rate of 500 mL/h, until right atrial pressure decreased to 50% of baseline, caused variable humoral, circulatory, and diuretic effects that were mainly related to the extent of fluid retention. In fact, in 10 patients (Group 1) with overhydration refractory to drug therapy and with urinary output less than 1,000 mL/24 h (mean, 370 mL), soon after the procedure, plasma renin (-39%), aldosterone (-50%), and norepinephrine (-47%) were reduced and RPP was increased (+16%), and in the subsequent 24 hours, diuresis was increased by 493%; in 9 patients (Group 2) whose baseline urinary output exceeded 1,000 mL/24 h (mean, 1,785 mL), renin increased by 40%, norepinephrine, aldosterone, and RPP each decreased by 12%, and diuresis remained unchanged; in 13 patients (Group 3) with a daily urinary excretion as in Group 2 and without overhydration, RPP decreased (-7%), renin (+196%), aldosterone (+170%), and norepinephrine (+52%) increased, and diuresis decreased by 45%. There was an overall correlation (p < 0.0001) between the combination of changes in these circulatory and hormonal variables and changes in diuresis and natriuresis with ultrafiltration. CONCLUSIONS: It appears that in CHF, (1) retention of sodium and water results from an interaction of hormonal and hemodynamic (primarily RPP) alterations that may exert a reciprocal positive feedback; (2) depending on the presence and severity of fluid retention, the response to withdrawal of body fluid may vary from neurohumoral activation and restriction of diuresis to neurohumoral depression and extreme potentiation of salt and water excretion; (3) refractory CHF requires the interruption of the humoral-hemodynamic vicious circle, and ultrafiltration is able to accomplish that. PMID- 8420300 TI - Risk of disease progression in asymptomatic multiple myeloma. AB - PURPOSE: In recent years, increasing numbers of patients with asymptomatic multiple myeloma have been diagnosed by chance and followed without any therapy. Those at risk for early or late disease progression should be distinguished in order to prevent complications. This study defined prognostic factors that would predict the need for early treatment. PATIENTS AND METHODS: We followed 95 patients with asymptomatic multiple myeloma without chemotherapy between 1974 and 1991. Magnetic resonance imaging (MRI) of the spine was conducted in 23% of patients with normal radiographs. An increase in serum myeloma protein to more than 50 g/L or new lytic bone lesions justified the institution of chemotherapy. Response to treatment and survival were assessed, and prognostic factors were defined for early or late disease progression by standard techniques. RESULTS: The median time to progression in all patients was 26 months. The 25 patients with either a lytic bone lesion, or both serum peak greater than 30 g/L and Bence Jones proteinuria, had the shortest median time to progression of 10 months; the 27 patients without any harmful factor remained stable for a median of 61 months. MRI confirmed bone or marrow disease in half of the patients with normal radiographs and may assist in the prognostic staging. Despite the markedly different times of disease stability, the response rates and survival after chemotherapy were similar for all groups of patients. CONCLUSION: Among asymptomatic patients with multiple myeloma, the extent of disease at diagnosis and the subsequent rate of disease evolution were major factors in the total survival time. These patients are a markedly heterogeneous group who may benefit from different approaches to treatment according to defined risk factors. PMID- 8420301 TI - Vitamin K and maintenance of skeletal integrity in adults. AB - PURPOSE: To determine the role of vitamin K status in the maintenance of skeletal integrity in adults. PATIENTS AND METHODS: 1. Bone mineral density (BMD) was measured by quantitative digital radiography (QDR) in 50 patients taking a vitamin K antagonist (warfarin) who were recruited from a large urban cardiology practice, and 50 age-, sex- and race-matched controls recruited from the community. 2. The relationship of BMD versus indices of vitamin K status (determined by measuring levels of vitamin K and descarboxyprothrombin in the plasma) in 113 nonanticoagulated adults was assessed. RESULTS: Measurements of BMD in the hip and spine were similar in anticoagulated subjects and matched controls. Multivariate analysis revealed that use of warfarin was not associated with a lower BMD. Ninety-five percent confidence intervals excluded a 0.06 g/cm2 reduction in BMD associated with the use of warfarin. Indices of vitamin K status did not correlate with BMD in normal subjects. CONCLUSIONS: Patients receiving long-term maintenance therapy with a vitamin K antagonist have normal bone density. BMD is unrelated to vitamin K status in nonanticoagulated subjects. These data suggest that vitamin K does not have a major role in maintenance of skeletal integrity in adults. PMID- 8420302 TI - Chronic myelogenous leukemia in the lymphoid blastic phase: characteristics, treatment response, and prognosis. AB - PURPOSE: To determine the clinical and laboratory characteristics and outcomes of patients with chronic myelogenous leukemia (CML) in the lymphoid blastic phase. PATIENTS AND METHODS: Data from 296 patients with CML blastic-phase disease who were referred to our institution between 1967 and 1991 were analyzed. Sixty-eight patients had CML lymphoid blastic-phase disease. Pretreatment characteristics, responses to different therapies, and survival rates were evaluated. RESULTS: Compared with patients having myeloid or undifferentiated blastic-phase disease, those with lymphoid morphology were significantly younger, presented with significantly lesser degrees of anemia, lower white blood cell and peripheral blast counts, higher percentages of marrow blasts, lower lactic dehydrogenase levels, and higher albumin levels. Accelerated-phase CML preceded the blastic phase in 40% of patients with lymphoid disease, compared with 54% of those with other morphologic findings (p = 0.03). The common acute lymphocytic leukemia antigen (CALLA) was expressed on lymphoid blasts in 97% of patients. The incidence of chromosomal abnormalities was similar in the three morphologic categories, although patients with lymphoid disease tended to have a lower incidence of trisomy 8 (17% versus 27%; p = 0.10) and of isochromosome 17 abnormalities (10% versus 17%; p not significant). The incidence of lymphoid blastic phase disease has not increased over the past decade, and it is not higher in patients with the chronic phase of the disease treated with alpha interferon. Patients with lymphoid disease had a significantly higher response rate to chemotherapy during the first salvage (49% versus below 20% for other morphologies; p < 0.001), particularly with vincristine, Adriamycin, and dexamethasone therapy (complete response rate of 61%). Survival during the blastic phase of the disease was also significantly longer in patients with lymphoid morphology than in those with other morphologies (median survival of 9 months versus 3 months; p = 0.01). The benefit associated with lymphoid blastic phase morphology is brief, and plans for allogeneic bone marrow transplantation or experimental maintenance or consolidation programs should be implemented rapidly. CONCLUSIONS: Patients with CML lymphoid blastic-phase disease have different clinical and laboratory features than patients with other blastic-phase morphologies. In patients developing CML blastic-phase disease, distinguishing those with lymphoid transformation is extremely important because of the different therapeutic requirements (acute lymphocytic leukemia-type therapy) and prognostic implications. PMID- 8420304 TI - Gordian knot. PMID- 8420303 TI - Effect of gemfibrozil in men with primary isolated low high-density lipoprotein cholesterol: a randomized, double-blind, placebo-controlled, crossover study. AB - PURPOSE: To evaluate the efficacy of gemfibrozil in men with primary isolated low high-density lipoprotein cholesterol (HDL-C) levels. PATIENTS AND METHODS: Fourteen men with low levels of HDL-C but desirable total cholesterol levels received gemfibrozil in a randomized, double-blind, placebo-controlled, crossover trial. The men were placed on a National Cholesterol Education Program Step-Two Diet. They were randomly assigned to receive placebo and gemfibrozil each for 3 months, with a 1-month washout period between phases. RESULTS: Overall, gemfibrozil increased the total HDL-C concentration by 9.2% (p = 0.001), reduced triglyceride (TG) levels by 38% (p < 0.01), and significantly lowered the total cholesterol:HDL-C ratio (p = 0.01). Those with fasting TG levels of 1.07 mmol/L (95 mg/dL) or greater had a significant elevation in the HDL-C level (14.6%, p = 0.005) and a reduction in TG levels (50%, p = 0.002) with gemfibrozil; those with fasting TG levels less than 1.07 mmol/L had a smaller increase in the HDL-C level (4.1%, p > 0.05) and a smaller reduction in TG levels (15%, p = 0.02). There were no significant differences in the plasma levels of low density lipoprotein cholesterol, HDL2-C, apolipoproteins (apo) A-I and B, or Lp(a). HDL3-C and apo A II levels rose slightly. The adverse effects attributable to gemfibrozil were minimal. CONCLUSION: In men with desirable total cholesterol levels, gemfibrozil raises HDL-C and lowers TG levels to a similar extent as reported for hyperlipidemic men in the Helsinki Heart Study. These lipid-altering effects were most pronounced in those with the highest fasting TG levels. PMID- 8420305 TI - Genetics of essential hypertension. AB - Blood pressure is a complex quantitative trait that is determined by multiple environmental and genetic factors. Although some simple Mendelian forms of high blood pressure have been described, essential hypertension is characterized by a complex mode of inheritance. Based on recent advances in molecular biology and statistical genetics, it has become feasible to search for chromosome regions that may contain genes contributing to the pathogenesis of hypertension in humans. For example, recent linkage and association studies have raised the possibility that a blood pressure regulatory locus may exist in or near the angiotensinogen gene on chromosome 1. Detailed genetic experiments in animal models of hypertension may help to guide further clinical studies and lead to an improved understanding of gene action in the pathogenesis of essential hypertension. PMID- 8420306 TI - Chronic meningitis in a 51-year-old man. PMID- 8420307 TI - Reporting the results of cystic fibrosis carrier screening. AB - The recent discovery of the cystic fibrosis gene has offered the possibility of population-based cystic fibrosis carrier screening. Although > 100 distinct mutations have been identified, five of these in aggregate represent about 85% of the alleles in Britain and the United States. Screening programs that test for these five mutations can be designed to offer several alternative ways to communicate the risk to a pregnancy and several alternative ways to manage a pregnancy. At this time we favor a strategy of screening partners in a couple in sequence, screening the second partner only if the first is positive; nevertheless, different strategies will appeal to different couples. PMID- 8420308 TI - Effect of prolonged oral terbutaline therapy on glucose tolerance in pregnancy. AB - OBJECTIVE: Our objective was to elucidate the pathophysiologic effects and potential reversibility of terbutaline-induced changes in carbohydrate metabolism. STUDY DESIGN: We prospectively evaluated serum glucose, insulin, glucagon, C-peptide, and pancreatic polypeptide levels in response to a 100 gm glucose challenge (oral 3-hour glucose tolerance test) in 17 obstetric patients without complications who were given terbutaline (5 mg orally every 4 hours) for 5 consecutive days between 24 and 32 weeks' gestation. Each patient served as her own control, with day 1 representing pretreatment, day 7 the treatment phase, and day 14 the posttreatment evaluation. Body mass index and posttreatment serum terbutaline levels were also measured. RESULTS: A significant initial treatment effect (day 1 versus 7) was observed for glucose (elevated), insulin (elevated), insulin/glucose ratio (elevated), and pancreatic polypeptide (elevated). A significant delayed treatment effect (day 1 versus 14) was also observed for insulin (elevated), insulin/glucose ratio (elevated), and pancreatic polypeptide (elevated). Body mass index directly correlated with postchallenge measures of insulin, insulin/glucose ratio, pancreatic polypeptide, and C-peptide. Posttreatment serum terbutaline levels directly correlated with pancreatic polypeptide, but not with other parameters. CONCLUSIONS: Our data support a dose independent, terbutaline-induced glucose intolerance mediated by glucagon and caused by diminished insulin sensitivity. PMID- 8420309 TI - Large fetal heart rate decelerations at term associated with changes in fetal heart rate variation. AB - OBJECTIVE: The objective was to determine whether large antepartum decelerations in fetal heart rate were associated with a switch from high to low fetal heart rate variation, suggestive of a change in sleep state, and whether the variation predicted outcome. STUDY DESIGN: Retrospective computerized analysis of 10,272 cardiotocographic records from 3998 patients at 37 to 42 weeks' gestation from four centers in England and Italy identified 140 good-quality records with large decelerations (more than 20 lost beats in area). RESULTS: In otherwise normal cardiotocograms a large deceleration had a 40% chance of association with a downward change in fetal heart rate variation (69% when the deceleration exceeded 100 lost beats). The change resembled that occurring naturally with behavioral states. Uterine contractions did not always precede large decelerations. When they did, the lag time (peak of contraction-trough of deceleration) increased from 28 seconds (at 20 to 29 lost beats) to > 100 seconds with increase in deceleration area. Of patients with large decelerations 76% had a normal vaginal delivery. CONCLUSION: Large decelerations near term, present in up to 5% of patients with otherwise normal fetal heart rate and variation, are often associated with a fall in fetal heart rate variation characteristic of a change in sleep state, without ominous significance. PMID- 8420310 TI - First-trimester ultrasonography findings in women with a history of recurrent pregnancy loss. AB - OBJECTIVE: We tabulated first-trimester ultrasonography findings in women with recurrent pregnancy loss and determined the rate of subsequent pregnancy loss and live births after demonstration of a live embryo. STUDY DESIGN: Sixty-seven women with three or more recurrent miscarriages underwent first-trimester ultrasonography and were prospectively followed through 101 pregnancies. RESULTS: First-trimester ultrasonography showed a dead embryo in seven pregnancies. Two of gestational sac in 12, and an empty uterus in four. A live embryo was seen in 78 pregnancies. Two of these pregnancies were terminated because of fetal chromosomal abnormalities. Of the remaining 76 pregnancies, 13 (17%) ended in a fetal loss (spontaneous abortion or fetal death) and 63 (83%) resulted in viable live births. CONCLUSIONS: Among women with recurrent pregnancy loss the presence of a live embryo detected by first-trimester ultrasonography is not as encouraging as in normal pregnant women. These findings are useful in counseling patients with recurrent pregnancy loss. PMID- 8420312 TI - The reliability of ultrasonography in the management of spontaneous abortion, clinically thought to be complete: a prospective study. AB - OBJECTIVE: The purpose of this study was to determine the reliability of ultrasonographic examination in the clinical management of spontaneous abortions thought to be complete. STUDY DESIGN: Patients with clinical findings suggestive of complete spontaneous abortion were managed prospectively on the basis of ultrasonographic results. Patients with an "empty uterus" were managed expectantly; those with a thickened endometrium or echoes consistent with retained tissue were subjected to curettage. RESULTS: Forty-eight of 49 (98%) patients with an "empty uterus" by ultrasonographic examination had an uneventful recovery without curettage. Nine of 13 (69%) patients with a thickened endometrium or evidence of retained tissue on ultrasonography had chorionic villi obtained by curettage. CONCLUSION: Ultrasonography is a highly reliable test in the management of women thought to have complete spontaneous abortion. PMID- 8420311 TI - Preventive effects of transdermal 17 beta-estradiol on osteoporotic changes after surgical menopause: a two-year placebo-controlled trial. AB - OBJECTIVE: The purpose of our study was to evaluate the effects of three dosages of transdermally administered 17 beta-estradiol on markers of bone loss in women who had recently undergone surgical menopause. STUDY DESIGN: This was a 2-year, randomized, double-blind, placebo-controlled study that reviewed 127 women stratified by age. Biochemical indicators of bone metabolism in urine and serum were periodically assessed, as was bone mineral content of the lumbar spine and radius. Statistical analysis examined the percent changes from baseline in bone mineral density by using an analysis of covariance with treatment as a factor and the baseline value as a covariant, by performing all-pairwise comparisons among the three estradiol groups, and by testing for a linear dose-response relationship. RESULTS: After 2 years of therapy, a significant dose-response relationship was detected. CONCLUSION: This 2-year study demonstrates that transdermally administered 17 beta-estradiol is a safe and effective regimen for preventing bone loss in recently postmenopausal women. PMID- 8420313 TI - Reduction of thromboxane A2 synthesis in pregnancy by polyunsaturated fatty acid supplements. AB - OBJECTIVE: High-dose supplements of fish oil reduce thromboxane synthesis in nonpregnant human subjects and were therefore proposed as a means of preventing various small-vessel disorders, including preeclampsia. The effect of fish oil on thromboxane metabolism in pregnancy was investigated in our study. STUDY DESIGN: Sixteen normal pregnant women in the third trimester of pregnancy were treated with a daily ingestion of 6 gm fish oil capsules containing 1.6 gm of n-3 fatty acid. In five patients the treatment was stopped because of severe-flavored reflux and hiccups. Eleven patients completed 3 weeks of treatment. Twenty-four hour urinary 11-dehydro-thromboxane B2 was measured by means of radioimmunoassay before and after completion of the study protocol in these 11 patients and in seven control pregnant women who did not receive the oil treatment. RESULTS: A decrease ranging from 32% to 71%, in 24-hour urinary 11-dehydro-thromboxane B2 excretion (mean reduction from 1606 pg/mg creatinine to 779 pg/mg creatinine, p < 0.001) was found among the 11 fish oil-treated women. No change in excretion was found among the control women. No maternal, fetal, or neonatal bleeding disturbances occurred, and no laboratory changes in coagulation markers were observed. CONCLUSIONS: High-dose n-3 fatty acid intake in pregnancy significantly reduces maternal thromboxane A2 synthesis. These results may provide a basis for a possible role of fish oil in managing patients at risk for preeclampsia. PMID- 8420314 TI - Two cases of multiple umbilical cord abnormalities resulting in stillbirth: prenatal observation with ultrasonography and fetal heart rates. AB - Two cases of multiple umbilical cord abnormalities consisting of three separate findings is presented. Both cases contained nuchal cords, knots, and umbilical cord thrombosis. One case had prenatal evidence of these findings by an ultrasonographic study videotaped at 32 weeks' gestation. When multiple umbilical cord abnormalities are diagnosed prenatally in the same fetus, that fetus should be considered to be at high risk; delivery should be considered at the least provocation. PMID- 8420315 TI - The PROEF diet--a new postoperative regimen for oral early feeding. AB - OBJECTIVE: The purpose of this study was to determine if immediate postoperative feeding of a new oral elemental diet (PROEF diet) would be tolerated by patients and to determine its effect on gastrointestinal function after cesarean section. STUDY DESIGN: One hundred eighteen patients undergoing cesarean section were randomly assigned by a computer-generated list of numbers to receive either the PROEF diet (60 patients) or routine postoperative dietary management (58 patients). Gastrointestinal morbidity was analyzed by an independent-samples t test. RESULTS: The PROEF diet group has a more rapid return of normal bowel sounds, 10.3 versus 14.5 hours (p = 0.001), and earlier acceptance of a regular diet, 2.0 versus 2.3 days (p = 0.008). CONCLUSION: The PROEF diet was well tolerated in cesarean section patients with no increase in gastrointestinal morbidity when compared with a control group of patients. This dispels the classic teaching that postoperative patients may not have oral intake until the return of normal bowel function. Further study is necessary to support the theoretic benefits that may accrue from early feeding of an elemental diet. PMID- 8420316 TI - Effect of ethanol and progesterone on monoamine oxidase activity in cultured cells of human term placenta. AB - OBJECTIVE: The aim of this study was to evaluate the effect of ethanol and progesterone on the monoamine oxidase activity in cultured human term placental cells. STUDY DESIGN: Human placental cells were prepared from normal human term placentas by enzymatic dispersion in Dulbecco's modified Eagle medium. The viability of placental cells prepared by our method was 90%, and the yield of placental cells was 0.6 x 10(6) cells per gram of wet placental tissue. Five milliliters of Dulbecco's modified Eagle medium containing 3 x 10(5) placental cells was plated in a 25 cm2 flask and cultured for 8 days in an incubator at 37 degrees C under an atmosphere of 5% carbon dioxide and 95% oxygen with a saturated humidity. During the culture period the culture medium was replenished every 2 days. A confluent monolayer condition was achieved after 8 days in culture. The cultured placental cells were treated with different concentrations of ethanol (0, 34.6, and 69.2 mmol/L) and progesterone (0, 16, and 32 mumol/L) on day 8 of culture for 48 hours. At the end of treatment placental cells from control and treated flasks were harvested for the analysis of monoamine oxidase activity by spectrophotometry. The effects of ethanol and progesterone on cultured placental cells were statistically analyzed by one-way analysis of variance followed by Duncan's multiple comparisons procedure. RESULTS: A human placental cell culture system has been established from normal human term placentas. The monoamine oxidase activity in 8-day-cultured human term placental cells was significantly higher than that of freshly prepared placental cells. Ethanol concentrations at 34.6 and 69.2 mmol/L significantly increased and progesterone concentration at 32 mumol/L significantly decreased the monoamine oxidase activity. CONCLUSION: These results suggest that the cultured human term placental cells can be used to examine the in vitro effects of ethanol and progesterone on monoamine oxidase activity. However, the physiologic significance of progesterone's inhibitory effect and the stimulatory effect of ethanol monoamine oxidase activity in the in vivo system have yet to be further investigated. PMID- 8420317 TI - Protein S deficiency in pregnancy: a case report. AB - Protein S deficiency is uncommon, may cause recurrent thrombosis, and may complicate pregnancy. A patient with protein S deficiency presented with a stillbirth followed by postpartum pulmonary embolism. She then had a successful pregnancy managed by anticoagulation and close fetal monitoring. PMID- 8420318 TI - Effects of methyldopa on uteroplacental and fetal hemodynamics in pregnancy induced hypertension. AB - OBJECTIVE: Our purpose was to study the effect of methyldopa on uteroplacental and fetal hemodynamics in women with pregnancy-induced hypertension. STUDY DESIGN: A prospective study of Doppler ultrasonographic blood flow data before and after 1 week of methyldopa treatment was conducted at the Department of Obstetrics and Gynecology, National University Hospital, Singapore, in 20 women (mean 35 weeks' gestation) with pregnancy-induced hypertension. The main outcome measures were maternal blood pressure, maternal and fetal heart rate, and blood velocity waveforms characterized by the pulsatility index in the fetal ascending aorta, middle cerebral artery, umbilical artery, and maternal uterine and arcuate arteries. Statistical evaluation was performed with t tests for paired observations. RESULTS: Maternal mean arterial blood pressure was reduced 9.7 mm Hg (95% confidence interval -13.8 to -5.6), and mean heart rate decreased 6.3 beats/min (95% confidence interval -11.1 to -1.4). Mean pulsatility index in the uterine (0.93 to 0.92) and arcuate arteries (0.61 to 0.73) were unchanged after treatment. Fetal hemodynamic changes before and after treatment were not significant. Fetal and neonatal outcome was uneventful. CONCLUSION: Short-term treatment with methyldopa in the last trimester in women with pregnancy-induced hypertension reduced maternal blood pressure and heart rate but had no adverse effects on uteroplacental and fetal hemodynamics. PMID- 8420319 TI - First-trimester simple hygroma: cause and outcome. AB - OBJECTIVE: First-trimester fetuses with simple nuchal hygromas represent a population that is different from midgestation nuchal cystic hygroma in terms of karyotype abnormalities and long-term prognosis. STUDY DESIGN: Sixty-eight consecutive fetuses diagnosed with simple nuchal hygromas between 10 and 14 weeks' gestation were evaluated ultrasonographically and karyotyped. Those with normal chromosome complements were ultrasonographically monitored throughout the remainder of the pregnancy for resolution of their hygromas. RESULTS: Twenty three of 27 fetuses with normal karyotypes resolved their hygromas within 4 weeks of diagnosis and were phenotypically normal at birth. Forty-one fetuses were karyotypically abnormal, with trisomy 21 being the most common abnormality. CONCLUSIONS: Fetuses with first-trimester simple nuchal hygromas are at high risk for aneuploidy and should be offered prenatal testing. Such fetuses with normal karyotypes will likely resolve their hygromas by 18 weeks' gestation, and most will be phenotypically normal at birth. PMID- 8420320 TI - Effect of age, parity, and smoking on pregnancy outcome: a population-based study. AB - OBJECTIVES: The purpose of our study was to investigate the combined interactive effects of maternal age, parity, and smoking on pregnancy outcome. STUDY DESIGN: This was a population-based Swedish study (n = 538,829). RESULTS: Multiple logistic regression analysis showed that the smoking-related effect on the relative increase in the odds ratio of low birth weight and preterm delivery was significantly greater among multiparous patients than nulliparous; among multiparas, smoking increased the odds ratios for low birth weight and preterm delivery by 2.4 and 1.6; the corresponding relative increases in the odds ratios among nulliparas were 1.7 and 1.1, respectively. With advancing maternal age there was a smoking-related relative increase in the odds ratios for small-for gestational-age births. Moreover, the age effect on the relative increase of low birth weight, preterm delivery, and small-for-gestational-age births was greater among nulliparas than multiparas. CONCLUSIONS: Older smokers are at an especially high risk for small-for-gestational-age births, and parous smokers are at an especially high risk for low birth weight and preterm delivery. PMID- 8420321 TI - Early transabdominal chorionic villus sampling in couples at high genetic risk. AB - OBJECTIVE: The purpose of the study was to evaluate the feasibility and safety of transabdominal chorionic villus sampling before 9 weeks' gestation. STUDY DESIGN: Two hundred pregnancies at risk for beta-thalassemia (n = 198) or Duchenne muscular dystrophy (n = 2) underwent transabdominal CVS at 6 through 8 weeks. Sampling success and fetal loss are expressed in percentages. RESULTS: Sampling was successful in all cases (100%). Forty-eight fetuses were affected by beta thalassemia and one by Duchenne muscular dystrophy. The percentage of fetal loss, expressed as a proportion of continuing pregnancies, was 4.0%. All women (n = 144) have been delivered, and no misdiagnoses have occurred. We observed one anencephalus and one mild limb defect consisting of absence of distal phalanges of index and little fingers of both hands and distal phalanges of both little toes. CONCLUSION: Transabdominal CVS before 9 weeks is a reliable and relatively safe method for prenatal diagnosis in patients at high risk for genetic diseases. However, further studies are necessary to assess the risk to the fetus. PMID- 8420322 TI - Fetal urine analysis for the assessment of renal function in obstructive uropathy. AB - OBJECTIVES: The assessment of fetal renal function plays a key role in the evaluation of posterior urethral valve obstruction cases. The aim of our study was to determine the value of several urinary compounds, including beta 2 microglobulin, N-acetyl-beta-D-glucosaminidase, and microalbumin in the assessment of prenatal renal function in cases of posterior urethral valve and their potential role in the selection of such cases for in utero shunting. STUDY DESIGN: A range of urinary compounds was measured, including beta 2 microglobulin, N-acetyl-beta-D-glucosaminidase, and microalbumin in 25 cases of posterior urethral valve obstruction. These cases were divided into four groups based on outcome. The Mann-Whitney test and analysis of covariance were used. RESULTS: Sodium, calcium, and beta 2-microglobulin were the best predictors for fetal survival. beta 2-Microglobulin values > 13 mg/L were almost invariably associated with fatal outcome. CONCLUSION: The estimation of beta 2-microglobulin may help in counseling parents and in selecting cases for in utero shunting. PMID- 8420323 TI - Leiomyosarcomas: clinical presentation. AB - OBJECTIVE: The ability to preoperatively distinguish benign from malignant myomas is increasingly important because of the availability of more conservative approaches to management of patients with benign uterine enlargement who wish to preserve fertility. Such therapies include observation, medical treatment with gonadotropin-releasing hormone analogs, and hysteroscopic resection of submucosal leiomyomas. In view of these options, this study was undertaken to identify characteristic features of leiomyosarcomas that may preoperatively distinguish benign from malignant myomas. STUDY DESIGN: The 10-year experience with 21 patients with uterine leiomyosarcomas at Yale-New Haven Hospital was retrospectively reviewed. RESULTS: Leiomyosarcomas were both broad-based and pedunculated and had no preferential uterine location. In 95% of the uterine specimens the leiomyosarcoma was either the largest or the only mass. The leiomyosarcoma was limited to one mass in all patients but one. CONCLUSION: We recommend that the largest myoma be monitored most closely during conservative management of a uterus containing leiomyomas. PMID- 8420324 TI - Functional asymmetry of the human myometrium documented by color and pulsed-wave Doppler ultrasonographic evaluation of uterine arcuate arteries during Braxton Hicks contractions. AB - OBJECTIVE: We hypothesized that arcuate arteries supplying placental and nonplacental myometrial portions would respond with different degrees of change in their resistance during Braxton Hicks contractions. STUDY DESIGN: We examined 20 healthy pregnant women between 18 and 24 weeks' gestation with pulsed-wave and color-flow Doppler during focal Braxton Hicks contractions identified by real time ultrasonography by means of the characteristic thickening of the myometrium. Systolic/diastolic ratio was used as an expression of resistance. Statistical analysis was performed by Wilcoxon signed-ranks and Mann-Whitney test. RESULTS: When the contractions are localized in the subplacental myometrium, the resistance of the arcuate artery did not differ during and after the contraction. In contrast, when the contraction involved only nonplacental myometrium, the resistance during the contraction was significantly higher and in some patients there was complete absence of flow during the diastolic phase. During subplacental myometrial contractions, the main uterine artery resistance was not affected. When the contraction involved the nonplacental myometrium, the resistance of the main uterine artery increased with more pronounced changes when the contraction involved the lateral myometrial wall ipsilateral to the uterine artery under examination. CONCLUSION: We speculate that the differences in the degree of resistance change are the result of different degrees of contractility exhibited by the subplacental and nonplacental myometrium. We conclude that the intact human myometrium manifests functional asymmetry and our Doppler findings confirm previous in vitro studies. PMID- 8420325 TI - Effect of fetal movement on the amniotic fluid index. AB - OBJECTIVES: Fetal movement has been shown to change the size and location of amniotic fluid pockets during measurement of the amniotic fluid index. The effect of redistributing the fixed intrauterine fluid volume on the amniotic fluid index is unknown. Therefore we tested the hypothesis that the amniotic fluid index was unaffected by fetal movement. STUDY DESIGN: A single examiner prospectively determined the amniotic fluid index before and after three discrete episodes of fetal movement during 96 biophysical profiles. A reliable blinded examiner provided a second postmovement measurement as a control. Data were analyzed by the paired t test. RESULTS: The mean change in the amniotic fluid index after fetal movement was 1.5 +/- 0.1 cm and 2.5 +/- 0.2 cm for postmovement determinations by the same examiner and blinded observer, respectively (p < 0.001). CONCLUSION: Interobserver and intraobserver variation can account for the change in the amniotic fluid index after fetal movement. PMID- 8420326 TI - A longitudinal Doppler ultrasonographic assessment of the alterations in peripheral vascular resistance of uterine arteries and ultrasonographic findings of the involuting uterus during the puerperium. AB - OBJECTIVE: Our purpose was to assess the alterations in peripheral vascular resistance of uterine arteries in the puerperium. STUDY DESIGN: In this longitudinal study 42 patients with uncomplicated pregnancies were examined with transabdominal color Doppler ultrasonography before and after delivery for 3 months. A total of 266 evaluations were performed. RESULTS: Within 2 days after delivery the mean pulsatility index of uterine arteries had increased significantly from 0.73 +/- 0.15 to 1.19 +/- 0.36, and the prediastolic notch had reappeared. After 6 weeks the indexes had started to increase further and reached a level of 1.75 +/- 0.56 by the end of the third puerperal month. CONCLUSIONS: Uterine vascular resistance increases soon after delivery. Puerperal changes in pulsatility indexes are triphasic. At the end of the fourteenth puerperal week pulsatility indexes are still much lower than those of nonpregnant women and all flow velocity waveforms represent a continuous end-diastolic component. PMID- 8420327 TI - Fetal heart rate response to sustained recreational exercise. AB - OBJECTIVE: We aimed to test the hypotheses that fetal heart rate increases during and after sustained exercise and that the magnitude of the increases is related to gestational age and the duration, intensity, and type of exercise. STUDY DESIGN: Maternal oxygen uptake and fetal heart rate were monitored in 120 regularly exercising women in association with routine 20-minute workouts between 16 and 39 weeks' gestation. RESULTS: In 97% of the studies fetal heart rate increased during and after exercise. This was significant at all gestational ages and with all forms of exercise with an overall increase of 15 +/- 11 beats.min-1 at 60% +/- 12% of maximal aerobic capacity (mean +/- SD). The magnitude increased with gestational age (10 +/- 8 to 20 +/- 11 beats.min-1), exercise intensity (8 +/- 7 to 21 +/- 13 beats.min-1), and exercise duration (8 +/- 4 to 16 +/- 7 beats.min-1). CONCLUSION: We concluded that the hypothesis is correct and speculate that these changes represent a maturing fetal response to a reduction in Po2. PMID- 8420328 TI - Induction of endothelial cell tissue factor activity by sera from patients with antiphospholipid syndrome: a possible mechanism of thrombosis. AB - OBJECTIVE: The hypothesis of our study is that antiphospholipid antibodies predispose to thrombosis by inducing endothelial cell tissue factor expression. STUDY DESIGN: Monolayers of cultured human umbilical vein endothelial cells were incubated for 8 hours in a medium containing 20% serum obtained either from patients with antiphospholipid antibodies (n = 11) or normal subjects (n = 8). Similar incubations were performed with immunoglobulin G fractions from either patients with antiphospholipid antibodies (n = 3) or normal subjects (n = 3). Endothelial cell tissue factor expression was measured with a tissue factor specific chromogenic substrate assay. The results were analyzed with a two-tailed t test. RESULTS: The mean endothelial cell tissue factor expression induced by antiphospholipid sera was significantly greater than the controls (p < 0.02). Immunoglobulin experiments indicated that the factor(s) responsible for the induction of tissue factor expression resides in the immunoglobulin G fraction of the sera (p < 0.01). CONCLUSIONS: Endothelial cell tissue factor expression is induced by antiphospholipid sera, with activity residing at least in part in the immunoglobulin fraction. The induction of tissue factor by antiphospholipid sera may play a role in the thrombotic tendency observed in patients with antiphospholipid syndrome. PMID- 8420329 TI - Prenatal diagnosis of a placental cyst: comparison of postnatal biochemical analyses of cyst fluid, amniotic fluid, cord serum, and maternal serum. AB - A 4 x 4 cm placental cyst was diagnosed prenatally by ultrasonography. At delivery, simultaneous samples of cyst fluid, amniotic fluid, cord blood, and maternal blood (collected 3 hours before delivery) were obtained. Biochemical measurements of cyst fluid, amniotic fluid, cord serum, and maternal serum suggest that the solutes of cyst fluid do not originate exclusively from a single source. In addition, X-cell synthesis and secretion into the cyst fluid may occur. PMID- 8420330 TI - Low-dose aspirin in the prevention of preeclampsia and fetal growth retardation: rationale, mechanisms, and clinical trials. AB - Preeclampsia is characterized by a functional imbalance between vascular prostacyclin and thromboxane A2 production. On the basis of the hypothesis that preeclampsia is at least partially caused by an increase in thromboxane A2, some studies attempted to correct this pathologic condition by pharmacologic manipulation with low-dose aspirin. The current literature suggests that the use of low-dose aspirin during pregnancy is safe with regard to congenital anomalies and fetal, neonatal, and maternal cardiovascular physiologic state and hemostasis. Aspirin at least partially corrects the pathologic increase in angiotensin II sensitivity that precedes the clinical development of preeclampsia. In addition, some clinical trials have demonstrated that low-dose aspirin is effective in reducing the incidence of preeclampsia and/or fetal growth retardation in selected high-risk women. Currently, large clinical trials are in progress to evaluate the effectiveness and side effects of the use of low dose aspirin in preventing preeclampsia and/or fetal growth retardation. Until these studies have been completed, it will remain unclear whether antiplatelet therapy, such as low-dose aspirin, should be adopted for the prevention of either preeclampsia or fetal growth retardation. PMID- 8420331 TI - Coitus late in pregnancy: risk of preterm rupture of amniotic sac membranes. AB - OBJECTIVES: Coitus with or without orgasm in late pregnancy is inconsistently associated with preterm rupture of amniotic sac membranes. We tested the hypothesis that during late pregnancy sexual behaviors including sexual positioning relate to the occurrence of premature rupture of membranes. STUDY DESIGN: Women aged 15 to 45 years having preterm premature rupture of membranes, term premature rupture of membranes, or preterm delivery without premature rupture of membranes were matched singly by age, race, and parity to control women delivered of term infants. Information about six sexual activities, obstetric history, cervical infections, smoking during pregnancy, and sociodemographic information was obtained by face-to-face interview. RESULTS: Only the male superior position was significantly associated with preterm premature rupture of membranes (odds ratio 2.40, 95% confidence interval 1.16 to 4.97) and preterm delivery without premature rupture of membranes (odds ratio 1.82, confidence interval 1.02 to 3.25) after confounding variables were controlled for. No sexual positioning or sexual activities related significantly to term premature rupture of membranes. CONCLUSION: Most sexual positions and activities during late pregnancy are not associated with adverse pregnancy outcomes. PMID- 8420332 TI - The effect of antibodies and immunotoxins reactive with HER-2/neu on growth of ovarian and breast cancer cell lines. AB - OBJECTIVE: Because HER-2/neu is overexpressed in one third of breast and ovarian cancers, we examined the effect of unconjugated monoclonal antibodies (ID-5, PB 3, TA-1) and an immunotoxin (TA-1-ricin) reactive with this protooncogene on the growth of breast and ovarian cancer cell lines. STUDY DESIGN: The tritiated thymidine incorporation assay was used to examine the effect of unconjugated antibodies on proliferation. A limiting dilution clonogenic assay was used to assess the effect of immunotoxin on cellular cytotoxicity. RESULTS: Scatchard analysis revealed that OVCA 420, OVCA 429, OVCA 432, and OVCA 433 cells had approximately 10(4) HER-2/neu receptors per cell, whereas the SKOv3 and SKBr3 cell lines expressed 10(5) and 10(6) receptors per cell, respectively. Monoclonal antibody ID-5 caused significant inhibition of tritiated thymidine incorporation in SKBr3, SKOv3, and OVCA 420 cells (p < 0.002). The TA-1-rich immunotoxin significantly inhibited the clonogenic growth of only SKBr3 and SKOv3 cells. CONCLUSION: HER-2/neu may be a useful target for immunotherapy with unconjugated antibodies and immunotoxins in ovarian and breast cancers that overexpress this protooncogene. PMID- 8420333 TI - Endothelin-1: immunocytochemistry, localization of binding sites, and contractile effects in human uteroplacental smooth muscle. AB - OBJECTIVE: Our purpose was to study the localization, distribution of binding sites, and contractile effects of endothelin-1 in human uteroplacental smooth muscle. STUDY DESIGN: The tissue localization of endothelin-1, the distribution of iodine 125-labeled endothelin-1 binding sites, and the mechanical effects of endothelin-1 were studied in isolated tissues from the human uterus and placenta by immunocytochemistry, autoradiography, and organ bath experiments. RESULTS: No specific endothelin-1 immunoreactivity could be detected in fetal placental tissues or in myometrium or intramyometrial arteries from term pregnant and nonpregnant women. In placental tissues a high density of iodine 125-labeled endothelin-1 binding sites was found in vessels of various sizes and in the chorionic villi, whereas the density in the jelly of Wharton was low. In myometrial tissue from pregnant and nonpregnant women a high density of iodine 125-labeled endothelin-1 binding sites was found, which in myometrium from pregnant women was mainly located to the myometrium and vascular smooth muscle. Endothelin-1 produced marked contractile responses in maternal and fetal uteroplacental vessels and in myometrial preparations. CONCLUSION: Endothelin-1 may be involved in the endogenous control of uteroplacental vascular and visceral smooth muscle. PMID- 8420334 TI - The angiotensin II antagonist saralasin inhibits ovulation in the perfused rat ovary. AB - OBJECTIVE: Our null hypothesis was that the angiotensin II antagonist saralasin does not reduce the number of ovulations in the rat ovarian perfusion model. STUDY DESIGN: Ovaries from pregnant mare's serum gonadotropin-stimulated immature rats were perfused with nutrient media to which luteinizing hormone and 3 isobutyl-1-methylxanthine had been added to induce ovulation. Test perfusions were treated with saralasin 1 mumol/L (n = 0.5) and compared with controls (n = 5) with the Student t test. Perfusions with both saralasin and angiotensin II and dose-response evaluations were performed. RESULTS: Saralasin-treated ovulations were 6.6 +/- 1.3 (mean + SEM) compared with 18.6 +/- 3.9, p < 0.02. The effects of saralasin could be reversed with the addition of an equimolar amount of angiotensin II. Dose-response evaluations showed a progressive inhibition of ovulation at 10(-8) to 10(-6) mol/L. CONCLUSION: The angiotensin II antagonist saralasin inhibits ovulation in a dose-dependent fashion; this effect is canceled by the addition of equimolar concentrations of angiotensin II. PMID- 8420335 TI - Phosphorus 31 magnetic resonance spectroscopy of perifused human placental villi under varying oxygen concentrations. AB - OBJECTIVE: Initial phosphorus magnetic resonance spectroscopy observations on the oxygen metabolism of placental villi from normal term pregnancies are described. STUDY DESIGN: Villi were suspended in medium and perifused within a custom designed 30 mm nuclear magnetic resonance probe in a superconducting vertical nuclear magnetic resonance magnet where pH, temperature, and oxygenation were monitored. RESULTS: Phosphorus resonances were observed from adenosine triphosphate, phosphomonoesters. inorganic phosphate, and phosphodiesters. No phosphocreatine signal was observed. The placental villus tissue responded to an increase in oxygen concentration of the perifusate with a rise in the adenosine triphosphate level and a concomitant decline in the inorganic phosphate and the phosphomonoester signals. CONCLUSION: The changes observed reflect continuing dynamic glycolysis and oxidative phosphorylation. The absence of a phosphocreatine peak suggests that aerobic pathways not driven by creatine kinase are important for placental metabolism. Our system demonstrates dynamic oxygen metabolism in perifused viable placental villus tissue by means of magnetic resonance spectroscopy. PMID- 8420336 TI - Expression and localization of matrilysin, a matrix metalloproteinase, in human endometrium during the reproductive cycle. AB - OBJECTIVE: We studied the expression of a matrix metalloproteinase, matrilysin, in the human endometrium to determine whether metalloproteinase genes are expressed during the reproductive cycle. Matrix metalloproteinases are a tightly regulated family of enzymes that degrade components of the extracellular matrix and basement membrane; they play important roles in growth and development and in invasion and metastasis of tumors and thus are likely enzymes participating in the dynamic structural changes occurring in endometrium during the reproductive cycle. STUDY DESIGN: In situ and Northern nucleic acid hybridization and immunohistochemistry were used to detect and localize matrilysin ribonucleic acid and protein in normal endometrial tissue. RESULTS: Matrilysin protein and matrilysin messenger ribonucleic acid are abundant in proliferative, late secretory, and menstrual endometrial epithelium but are not detected in early or mid secretory endometrium. CONCLUSION: The expression of the matrilysin gene is regulated in endometrium during the reproductive cycle, implying an important role for matrilysin in endometrial physiologic characteristics. PMID- 8420337 TI - Hemodynamic and fluid responses to furosemide infusion in the ovine fetus. AB - OBJECTIVES: The direct effects of furosemide infusion have not been systematically examined in the fetus. Our objective was to explore the hemodynamic and urinary responses to a 4-hour infusion of furosemide into fetal sheep. STUDY DESIGN: Furosemide (0, 1, 5, or 10 mg/hr) was infused into the fetal inferior vena cava for 4 hours in 15 chronically catheterized near-term sheep. RESULTS: Relative to vehicle infusion, furosemide produced dose-dependent increases in fetal arterial pressure (analysis of variance, p < 0.05 when comparing groups), fetal heart rate (p < 0.0001), urine flow (p < 0.0001), and urine osmolality, sodium, and chloride, concentrations (p < 0.0001). Concomitantly, there were dose-dependent decreases in fetoplacental blood volume, fetal plasma chloride (p < 0.0001) and fetal venous pressure (p < 0.05). The changes in urine flow rate and fetal arterial pressure were positively correlated (r = 0.46, p < 0.01), suggesting that the diuresis was mediated in part by fetal arterial pressure. The four-quadrant amniotic fluid index increased during the furosemide infusions but not during vehicle infusions (p = 0.022). CONCLUSIONS: Furosemide infusion caused a marked dose-dependent diuresis in the ovine fetus that appears to be partly mediated by increases in vascular pressure. Although amniotic fluid volume increases and fetal blood volume decreases, the reduction in blood volume was small compared with the urine volume excreted. PMID- 8420338 TI - Pregnancy-induced changes in the three-dimensional mechanical properties of pressurized rat uteroplacental (radial) arteries. AB - OBJECTIVE: The purpose of this study was to describe the effects of pregnancy on the size and three-dimensional mechanics of uterine radial arteries. STUDY DESIGN: Measurements of lumen diameter, wall thickness, and axial and radial distensibility were made in situ and in pressurized segments of excised vessels from nonpregnant (n = 29) and late-pregnant (days 19 to 21, n = 19) Sprague Dawley rats. RESULTS: In the unpressurized state, the overall length of the radial artery segment of the arcade increased approximately 4.8 times during gestation. Lumen diameter increased by 60%, as did distensibility in both the radial and axial directions. However, there was no measurable change in the thickness of the vascular wall, which was muscular in appearance and comprised approximately two layers of circumferentially oriented smooth muscle, a relatively thick internal elastic lamina, and a well-defined endothelial layer. Cross-sectional area increased significantly during pregnancy (1.37 times at 50 mm Hg), as did the overall volume of the vascular wall (6.86 times at 50 mm Hg), primarily as a result of arterial growth in the longitudinal (axial) direction. CONCLUSIONS: Remodeling of the radial artery segment of the uterine vasculature clearly occurs during gestation, resulting in vessels that are of a larger caliber and are longer and more distensible in both the axial and radial directions. PMID- 8420339 TI - Microinvasion of the amniotic cavity increases the risk of post-cesarean section endometritis. PMID- 8420340 TI - Doppler ultrasonography of human fetal ductus venosus. PMID- 8420341 TI - Fetal choroid plexus cysts. PMID- 8420342 TI - Pregnancy reduction in Jewish law. PMID- 8420343 TI - Plea for continuation of antenatal steroids at 24 to 28 weeks' gestation. PMID- 8420344 TI - Effects of monophasic low-dose oral contraceptives on fibrin formation and resolution in young women. AB - OBJECTIVE: The purpose of this study was to examine key variables in the regulation of coagulation and fibrinolysis during intake of low-dose oral contraceptives containing newly developed progestogens. STUDY DESIGN: Thirty-four healthy young women were allocated to 12 consecutive cycles of treatment with monophasic combinations of 20 micrograms ethinyl estradiol and 150 micrograms desogestrel (n = 15) or 30 micrograms ethinyl estradiol and 75 micrograms gestodene (n = 19). Nonparametric analysis of variance was used for statistical evaluation. RESULTS: In both groups plasma levels of fibrinogen and Factor VIIc increased, and the capacity of coagulation inhibition was affected by increased protein C and decreased protein S levels. Increased fibrinolytic capacity was indicated by elevated activity and reduced antigen levels of tissue plasminogen activator and by reduced activity and concentration of plasminogen activator inhibitor. Thrombin-antithrombin III complexes and fibrin degradation products were unchanged, signifying no effect of hormonal intake on the degree of activation of the coagulation system or the efficacy of fibrinolysis. CONCLUSION: The overall dynamic balance between generation and resolution of fibrin was maintained during treatment with both hormonal compounds. Our findings suggest that the risk of thrombosis in normal women should not be increased. PMID- 8420345 TI - Rapid intrapartum detection of group B streptococcal colonization with an enzyme immunoassay. AB - OBJECTIVE: The purpose of this study was to determine the accuracy of a rapid immunoenzyme assay for detecting intrapartum colonization with group B streptococci. STUDY DESIGN: Three rayon-tipped swabs were used to collect specimens from the posterior vaginal wall of 424 preterm and term patients in labor. Three tests were performed on specimens obtained from the first 182 patients: semiquantitative culture on blood agar, culture in selective Todd Hewitt broth, and ICON Strep B (Hybritech, San Diego) immunoconcentration assay. For the remaining 242 patients, the ICON test was performed only when the Todd Hewitt broth culture was positive. RESULTS: The prevalence of positive cultures was 23%. For the first 182 patients, the immunoassay had a sensitivity of 11%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 78%. The overall sensitivity for all 424 patients was 9%. In eight women with heavy colonization, defined as an inoculum of > 10(5) cfu/ml the sensitivity of the assay was 100%. In the study population there were three cases of group B streptococcal sepsis in infants whose mothers were only lightly colonized. None of these cases of colonization were detected by the assay. CONCLUSION: The ICON immunoconcentration assay is very sensitive in identifying heavy group B streptococcal colonization of > 10(5) cfu/ml but quite insensitive in detecting lower levels of colonization. Thus it is not a suitable test for general screening. PMID- 8420346 TI - The development of abnormal heart rate patterns after absent end-diastolic velocity in umbilical artery: analysis of risk factors. AB - OBJECTIVES: Our objectives were to evaluate the time interval elapsing between the occurrence of absent end-diastolic velocity in the umbilical artery and either the development abnormal fetal heart rate patterns or delivery and to establish the maternal and fetal factors that may affect this interval. STUDY DESIGN: Thirty-seven fetuses free of structural and chromosomal abnormalities in which the development of absent end-diastolic velocity in umbilical artery was evidenced by serial Doppler recordings were studied. At the first occurrence the following factors were considered: gestational age, presence of hypertension or preeclampsia, amniotic fluid index, severity of growth retardation, and 10 different Doppler indices calculated from extracardiac and intracardiac vascular districts. Actuarial statistical methods were applied, with the occurrence of antepartum late heart rate deceleration as the censoring variable. RESULTS: The interval between the first occurrence of absent end-diastolic velocity in umbilical artery and delivery ranged from 1 to 26 days. Indications for delivery were the development of antepartum late heart rate decelerations in 23 fetuses (62.1%) and different maternal or fetal complications in the remaining 14 fetuses. Multivariate analysis revealed that gestational age and the presence of hypertension and pulsations in umbilical vein were the dominant factors in determining the length of this time interval. CONCLUSION: The duration of the time interval between the occurrence of absent end-diastolic velocity in umbilical artery and abnormal heart rate pattern differs considerably among fetuses, and it is mainly determined by gestational age and presence of maternal hypertension and pulsations in umbilical vein. PMID- 8420347 TI - Platelet-activating factor-acetylhydrolase activity in normotensive and hypertensive pregnancies. AB - OBJECTIVE: The purpose of our study was to evaluate the hypothesis that pregnancy is associated with decreased platelet-activating factor-acetylhydrolase activity in women with normotension, but not in women with hypertension. STUDY DESIGN: We evaluated plasma platelet-activating factor-acetylhydrolase activity in normal nonpregnant women (n = 10), normal pregnant women (n = 24), pregnant women with pregnancy-induced hypertension-preeclampsia (n = 7), and a group of men with normotension (n = 10). RESULTS: Platelet-activating factor-acetylhydrolase activity was lower at 32 weeks of gestation during normal pregnancies compared with nonpregnant controls (p < 0.001); however, in women with pregnancy-induced hypertension-preeclampsia, platelet-activating factor-acetylhydrolase activity was not decreased. Platelet-activating factor-acetylhydrolase activity in men was higher than in all women (p < 0.01). CONCLUSION: Pregnant women with normotension may be refractory to pressor agents such as angiotensin II in part because of the decrease in plasma platelet-activating factor-acetylhydrolase activity, which results in an increase in platelet-activating factor. In contrast, enzyme activity is not decreased in pregnant women with hypertension, who have increased sensitivity to various pressor agents. PMID- 8420348 TI - A comparison of the Pipelle device and the Vabra aspirator as measured by endometrial denudation in hysterectomy specimens: the Pipelle device samples significantly less of the endometrial surface than the Vabra aspirator. AB - OBJECTIVE: The distribution and relative area of endometrial sampling of two endometrial biopsy techniques, Pipelle and Vabra aspiration biopsy, were compared. STUDY DESIGN: Twenty-five patients scheduled for hysterectomy were randomly assigned to undergo preoperative endometrial biopsy by Pipelle device (12 patients) or Vabra aspiration (13 patients). The endometrial cavities of the surgical specimens were photographed, and percent denudation was determined by a point-counting method. RESULTS: The percentage of endometrial surface area sampled by the Pipelle device was 4.2% +/- 0.92% (mean +/- SEM), and by Vabra aspirator 41.6% +/- 5.7% (p < 0.0001). The mean number of endometrial surfaces (one anterior and one posterior) sampled by the Pipelle device was 1.08 +/- 0.15, and by the Vabra aspirator 2 +/- 0 (p = 0.001). The mean number of endometrial quadrants (four anterior plus four posterior) sampled by the Pipelle device was 2.4 +/- 0.41, and by the Vabra aspirator 7.4 +/- 0.42 (p < 0.0001). CONCLUSION: We conclude that the Vabra aspirator is statistically superior to the Pipelle device for sampling the endometrial cavity when the total surface area sampled and the distribution of surfaces sampled are considered. PMID- 8420349 TI - Diagnosis of fetal infection in the patient with an ultrasonographically detected abnormality but a negative clinical history. AB - Unsuspected fetal infection (viral or parasitic) is often overlooked as a possible cause of abnormalities revealed on ultrasonographic examination during pregnancy. Infectious agents can produce a wide spectrum of ultrasonographic findings. Our objective was to highlight those findings and review the rationale and method of antenatal investigation. No ultrasonographic findings are pathognomonic for a particular agent. The search for possible fetal infection consists of both direct and indirect techniques. Methods to directly demonstrate fetal infection include maternal-fetal serologic studies, culture, electron microscopy, and polymerase chain reaction. Emerging evidence strongly suggests that prior assumptions concerning the sensitivity of both maternal and fetal neonatal serologic studies are incorrect. Serologic studies alone are inadequate to exclude fetal infection as a cause of an ultrasonographically detected abnormality. Indirect techniques suggestive of fetal infection include fetal hematologic and biochemical measurements. New approaches and methods for the detection of fetal infection are needed. The evaluation should be initiated antenatally. Needless delay until after delivery increases the likelihood that the diagnosis will be missed. Antenatal diagnosis provides the opportunity for therapy and often leads the practitioner to modify the obstetric and neonatal care plan. A protocol used by the University of Iowa Fetal Diagnosis and Treatment Unit for the diagnosis and management of unsuspected fetal infection is offered as an example of a protocol that has been successful in the past. PMID- 8420350 TI - The experience with vaginal birth after cesarean delivery in a small rural community practice. AB - OBJECTIVE: The purpose of this study was to determine if the success and safety of vaginal birth after cesarean delivery in a small, isolated, rural hospital compare with those seen in larger centers. STUDY DESIGN: As part of a continuing study, the prenatal and hospital records of all patients with previous cesarean delivered by the Rural Health Group between October 1988 and January 1991 were reviewed. Patients were allowed a trial of labor with one or more previous cesarean sections, unknown scar, and breech presentation but not for other malpresentation or a vertical scar. RESULTS: A total of 67 patients were studied; 76.1% of these had a trial of labor, and 60.8% of them were delivered vaginally, whereas 39.2% underwent repeat cesarean delivery. Of the 67 patients 11.9% were not candidates for vaginal birth after cesarean delivery, and the same percentage refused. Forty-nine percent received oxytocin; of these, 56% were delivered vaginally. Overall, maternal complications were similar between the groups. Two uterine ruptures occurred; neither was associated with labor. The major maternal complications occurred in the vaginal birth after cesarean delivery group, but all were associated with antepartum conditions and not related to labor and delivery. There were no maternal deaths. The only neonatal death resulted from a congenital anomaly. CONCLUSION: We concluded that vaginal birth after cesarean delivery can be performed safely in an isolated small community hospital with success rates similar to those of larger centers and with no increased maternal or neonatal morbidity or mortality. PMID- 8420351 TI - Intrapartum fetal heart rate assessment: monitoring by auscultation or electronic means. AB - OBJECTIVE: Our purpose was to assess the frequency with which auscultation could be used as the primary mode of fetal assessment during labor in a busy labor and delivery suite by means of published criteria. STUDY DESIGN: During a 3-month period, 862 patients in labor with live fetuses between 24 and 43 weeks of gestation were available for auscultation in the prospective study. Auscultation was initiated during a contraction and extended for 30 seconds after uterine activity ceased. It was repeated every 15 minutes in the first stage and every 5 minutes in the second stage of labor. RESULTS: In 420 patients this modality was not begun because of inability of the nurses to meet 1:1 staffing requirements. In 19 patients auscultation was not performed because of obesity (12) or patient refusal (7). Of the 423 assessed by auscultation 392 were unable to complete monitoring caused by the frequency requirement (n = 212) or the recording criteria (n = 163). Of the 31 patients where auscultation was successfully completed, there was a 1:1 nurse ratio during the entire labor. CONCLUSIONS: Auscultation with stringent evaluation and recording frequency is not feasible under normal labor and delivery room conditions unless 1:1 nursing care is always available. PMID- 8420352 TI - Urinary red blood cell and cast excretion in normal and hypertensive human pregnancy. AB - OBJECTIVE: Although the well-recognized pregnancy changes in renal blood flow, cellular function, tubular fluid composition, and flow rates may cause altered excretion rates of formed elements in the urine, relatively little attention has been paid to this in normal pregnancy. The urinary white cell excretion is known to be increased, with loss of the usual relationship between pyuria and urinary infection. Whether the same is true for other formed elements is unknown. Our purpose was to determine normal values for excretion of erythrocytes and casts and to establish whether they became abnormal in women in whom preeclampsia developed. STUDY DESIGN: Study subjects were 174 continuously normotensive pregnant volunteers, 22 women who had preeclampsia, and 8 women who had chronic essential hypertension. Early-morning midstream urine samples were collected at 17 to 20 and at 33 to 36 weeks' amenorrhea after insertion of a vaginal tampon. Urinary microscopy was performed by standard techniques (Kesson et al. Lancet 1978;2:809). RESULTS: The 95th percentile for concentration of erythrocytes was < or = 2500 red blood cells/ml; for casts the concentration was < or = 30 red blood cells/ml in normal pregnancy. Values for those who had preeclampsia and for the small group with chronic essential hypertension were within these limits. CONCLUSION: In an early-morning concentrated urine sample, values of < or = 2500 erythrocytes/ml and < or = 30 hyaline or granular casts per milliliter can be accepted as normal. PMID- 8420353 TI - Expression of ras oncogene p21 protein in normal and neoplastic ovarian tissues: correlation with histopathologic features and receptors for estrogen, progesterone, and epidermal growth factor. AB - OBJECTIVE: The aim of the study was to investigate the biologic significance of p21 expression in normal and neoplastic ovarian tissues. STUDY DESIGN: Western blotting analysis of p21/ras oncoprotein was conducted in a group of 14 normal and cystic ovaries, six benign tumors, 42 primary ovarian cancers, and 15 omental metastases. RESULTS: Levels of p21 were similar in normal and cystic ovaries and in benign tumors, whereas they were significantly higher in malignant tumors than in control tissues (median 1.91, range 0.12 to 5.00 vs median 1.03, range 0.32 to 2.20; p = 0.023) and in omental metastases than in primary ovarian carcinomas (median 3.05, range 0.55 to 5.72 vs median 1.97, range 0.12 to 5.00; p = 0.14). We found no correlation between p21 expression and histopathologic or clinical characteristics. Estrogen receptor-positive and progesterone receptor-positive tumors expressed higher p21 levels than did estrogen receptor-negative and progesterone receptor-negative tumors (p < 0.05), but no correlation with epidermal growth factor receptor status was found. In the univariate analysis of survival p21 positivity showed a negative prognostic value. CONCLUSION: The enhancement of p21 protein is associated in the ovarian tissue with the malignant phenotype and the acquisition of metastatic potential. PMID- 8420354 TI - A multicenter study of preterm birth weight and gestational age-specific neonatal mortality. AB - OBJECTIVE: This analysis was performed to present updated neonatal mortality data by age and birth weight for preterm newborns and to demonstrate the influence of plurality, ethnicity, and infant sex on mortality. STUDY DESIGN: Preterm birth weight and gestational age-specific mortality rates were compiled from the five centers that participated in the March of Dimes Multicenter Preterm Birth Prevention Project. In each center gestational age was assessed by standardized methods. A birth weight and gestational age-specific mortality chart for preterm births was created with live-birth data. RESULTS: In each birth weight group mortality decreased as the gestational age advanced; for each gestational age group heavier infants had less mortality. Female infants < 29 weeks survived better than male infants, and singletons < 29 weeks survived better than twins. Survival for black preterm newborns was better than that of whites but differences were not significant. Mortality for black term infants was significantly higher. The largest improvement in survival occurred between 25 and 26 weeks. At 30 weeks survival was > 90% and improved < 1% per week thereafter. CONCLUSIONS: When compared with rates in previous reports, mortality rates appear to have improved, especially at gestational ages < 29 weeks. These data may be useful in decision-making and in counseling patients at risk for preterm delivery. PMID- 8420355 TI - Increased thromboxane formation in diabetic pregnancy as a possible contributor to preeclampsia. AB - OBJECTIVE: Because pregnant women with diabetes have an increased risk of preeclampsia, we tested the hypothesis that urinary excretion of thromboxane metabolites is increased in diabetic pregnancies without evidence of preeclampsia at the time of testing. STUDY DESIGN: Urinary excretion of thromboxane A2 metabolites (either 2,3-dinor-thromboxane B2 or 11-dehydro-thromboxane B2) was measured in 24 type I pregnant diabetic individuals and in 20 women with normal pregnancies between 28 and 32 weeks' gestation. RESULTS: The amount of 2,3-dinor thromboxane B2 and 11-dehydro-thromboxane B2 in the urine of pregnant women with diabetes (1727 +/- 415 and 827 +/- 276 pg/mg creatinine) was significantly higher than in women with normal pregnancies (638 +/- 218 and 178 +/- 145 pg/mg creatinine) (p < 0.002 and p < 0.001). CONCLUSION: Our findings support a role for thromboxane in the pathogenesis of preeclampsia. PMID- 8420356 TI - Fetal plasma erythropoietin in pregnancies complicated by maternal diabetes mellitus. AB - OBJECTIVE: Our purpose was to investigate the relationship between fetal plasma erythropoietin concentration and measures of short-term and long-term glycemic control and fetal oxygenation in pregnancies complicated by maternal diabetes mellitus. STUDY DESIGN: A cross-sectional study was performed at The Harris Birthright Research Centre for Fetal Medicine, London. Cordocentesis was performed in 31 diabetic pregnancies for the measurement of umbilical venous blood pH, PO2, PCO2, lactate and glucose concentration, erythroblast count, hemoglobin, and plasma erythropoietin concentrations. RESULTS: The mean pH was significantly lower and the PCO2, lactate, erythropoietin, hemoglobin, and erythroblast counts were significantly higher than the appropriate normal mean for gestation. There were significant associations between (1) fetal erythropoietin and erythroblast count, (2) fetal erythroblast count and hemoglobin, (3) fetal hemoglobin and maternal glycosylated hemoglobin, and (4) maternal glucose and fetal glucose, pH, and lactate. CONCLUSIONS: We postulate that maternal hyperglycemia causes fetal hyperglycemia and acidemia. Increased erythropoietin may be caused by tissue hypoxia or hyperinsulinemia. The increase in fetal hemoglobin may be the consequence of increased erythropoiesis, mediated by either erythropoietin or hyperinsulinemia. PMID- 8420357 TI - Placental interleukin-6 production is enhanced in intrauterine infection but not in labor. AB - OBJECTIVE: Because interleukin-6 is an important mediator in the host defense mechanism against infection and tissue damage, we studied the capacity of placentas with or without either labor or chorioamnionitis in the third trimester to produce interleukin-6. STUDY DESIGN: The placental blocks were cultured, and their interleukin-6 titers were measured by a bioassay. RESULTS: Placentas with labor produced a similar amount of interleukin-6 to placentas without labor. In contrast, placentas with chorioamnionitis produced much more interleukin-6 than the placentas with or without labor (p < 0.0001). CONCLUSION: Placental interleukin-6 is thus surmised to participate in potentiation of the placental and fetomaternal defense mechanisms together with placental interleukin-1 during chorioamnionitis. PMID- 8420358 TI - Prenatal ultrasonographic diagnosis of intracranial teratoma and massive craniomegaly with associated high-output cardiac failure. AB - Congenital intracranial teratomas are rare and usually fatal. We present prenatal diagnosis of such a case associated with scalp, facial, and body skin edema, hepatomegaly, extramedullary hematopoiesis, polyhydramnios, and a hydropic placenta. These manifestations of high-output cardiac failure were thought to be the result of the large cardiac output required by massive intracranial tumor arteriovenous shunting. PMID- 8420359 TI - Traction retinal detachment. XLIX Edward Jackson Memorial Lecture. AB - This review had two main objectives. The first goal was to attempt to define a final common pathway for traction retinal detachment. To illustrate this, I considered the following three common clinical entities: penetrating ocular trauma, proliferative vitreoretinopathy, and proliferative diabetic retinopathy. My second goal was to emphasize the role of research in developing a rational approach to treatment. To illustrate the second goal, I reviewed the contributions from three realms of research--laboratory, animal, and clinical research--examining their interactions and considering their roles in our current understanding and management of these clinical entities. PMID- 8420360 TI - Multiple mitochondrial DNA deletions in myo-neuro-gastrointestinal encephalopathy syndrome. PMID- 8420361 TI - Subconjunctival silicone oil after vitreoretinal surgery. PMID- 8420362 TI - A diode laser mounted on a slit lamp for ophthalmic photodynamic applications of phthalocyanine. PMID- 8420363 TI - Unesterified cholesterol in granular, lattice, and macular dystrophies. PMID- 8420364 TI - Ciprofloxacin therapy of Mycobacterium chelonae keratitis. PMID- 8420365 TI - Aqueous humor protein and intraocular lens powers. PMID- 8420366 TI - Distinguishing compensatory head turn from gaze palsy in children with unilateral oculomotor or abducens nerve paresis. PMID- 8420367 TI - Tricholemmal carcinoma of the eyelid. PMID- 8420368 TI - Meibomian gland function and giant papillary conjunctivitis. PMID- 8420369 TI - Meibomian gland function and giant papillary conjunctivitis. PMID- 8420370 TI - Visual outcome after penetrating keratoplasty with double continuous or combined interrupted and continuous suture closure. PMID- 8420371 TI - Vagal responses to adjustable sutures in strabismus correction. PMID- 8420372 TI - Superior rectus muscle overaction after cataract extraction. PMID- 8420373 TI - Photoreceptor outer segments in the aqueous humor of patients with atopic dermatitis and retinal detachment. AB - We examined the aqueous humor by electron microscopy of four patients with atopic dermatitis and retinal detachment. All the patients had a moderate number of cells in the aqueous humor. Cataract impaired fundus visualization except for one patient with aphakia. A flat retinal detachment involved a wide area detected by echography. The intraocular pressure in the eyes with retinal detachment demonstrated a marked diurnal fluctuation compared with that of the fellow eye. The aqueous cells consisted predominantly of photoreceptor outer segments. This finding does not support the theory that aqueous cells in atopic dermatitis are of inflammatory origin. Photoreceptor outer segments in aqueous humor are indicative of rhegmatogenous retinal detachment in patients with atopic dermatitis. PMID- 8420374 TI - Serum levels of interleukin-2 receptor in ocular Behcet's disease. AB - Serum interleukin-2 receptor levels were evaluated in 69 patients who had chronic bilateral uveitis and in 22 control subjects. Fifty-one of the 69 patients with uveitis had the ocular type of Behcet's disease and 18 had pars planitis (intermediate uveitis). The mean serum interleukin-2 receptor level was 412.6 +/- 94.6 U/ml for the control group, 465.0 +/- 96.6 U/ml for the patients with intermediate uveitis, and 810.9 +/- 369.3 U/ml for those with ocular Behcet's disease. The serum interleukin-2 receptor levels of patients with ocular Behcet's disease were significantly different from the levels of both the control and the intermediate uveitis groups (P < .001). The differences in serum levels of patients with intermediate uveitis and the levels of the control subjects were not statistically significant. Treatment of patients with ocular Behcet's disease for four to six weeks with either cyclosporine, methylprednisolone, or a combination of both, decreased the intraocular inflammation in nearly all cases. The influence of treatment on the level of serum interleukin-2 receptor, however, was variable. PMID- 8420375 TI - Determination of corneal image-forming properties from corneal topography. AB - Keratometry provides useful information about the cornea's image-forming properties, such as corneal astigmatism, but is inaccurate on irregular corneas. Quantitative corneal topographic information is now obtainable on irregular corneas, but is difficult for the clinician to interpret. We developed a method to determine the spherical power, astigmatism, and topographic irregularity of a cornea by finding the best-fit spherocylinder that was closest to its measured topography. Keratometric measurements and two videokeratographs were gathered prospectively on 262 normal and abnormal corneas. The best-fit measurements of spherical power, astigmatism, and topographic irregularity were reproducible with one standard deviation of 0.75 diopter or better; agreement with keratometric measurements in normal eyes was good (0.60 diopter or better). Topographic irregularity averaged 0.1 diopter on precision spheres, 0.4 diopter on 146 normal eyes, 0.8 diopter on 29 eyes after radial keratotomy, 2.0 diopters on 58 eyes after penetrating keratoplasty, and 3.0 diopters on 29 eyes with advanced keratoconus. We conclude the following: basic corneal image-forming properties can be measured from videokeratographs; the properties can be determined, by our methods, on irregular corneas in which keratometry is unreliable; and topographic irregularity provides a measure of irregular astigmatism. PMID- 8420376 TI - Eyelid involvement in acanthosis nigricans. AB - We treated a 56-year-old man who had acanthosis nigricans associated with gastric adenocarcinoma and involving the axillae, hands, feet, anus, soft palate, and eyes. The patient developed epiphora from occlusion of the canaliculi with papillomatous lesions. Recognition of the ocular signs of this paraneoplastic syndrome may indicate to the ophthalmologist that the patient should have a thorough oncologic examination. PMID- 8420377 TI - Orbital meningoencephalocele manifesting as a conjunctival mass. AB - Orbital meningoencephalocele is a rare congenital abnormality caused by a defect of the cranio-orbital bones that usually manifests soon after birth as a soft, cystic fullness in the superomedial canthal area with associated exophthalmos. We managed an unusual case of congenital orbital meningoencephalocele that manifested as a cystic conjunctival mass without proptosis or periorbital changes. Preoperative computed tomographic scans failed to demonstrate a bone defect. After suture ligature of the posterior stalk, excision of the lesion yielded an ependymal cyst surrounded by neuroglial and meningeal tissue and filled with cerebrospinal fluid. The patient had normal results of ophthalmic and neurologic examinations after transconjunctival resection of the lesion after three years of follow-up. An orbital approach may be appropriate for a few selected cases in which no bone defect is found on computed tomography. Orbital meningoencephalocele should be included in the differential diagnosis of isolated congenital conjunctival cystic masses. PMID- 8420378 TI - Fluorescein and indocyanine green angiographies of central serous choroidopathy by scanning laser ophthalmoscopy. AB - We examined 19 patients (41 +/- 7.5 years old) with central serous chorioretinopathy and symptoms that ranged from one day to 24 months. Fluorescein and indocyanine green angiographies were performed with a scanning laser ophthalmoscope. Focal exudation was found in all patients with fluorescein and in 15 patients (79%) with indocyanine green. We found a more widespread exudation of indocyanine green into the choroid around the focal hyperfluorescent spot in seven patients (37%). Perfusion with fluorescein was delayed in the area of focal exudation in two patients (11%) and with indocyanine green in 12 patients (63%). Exudation of both dyes stopped with clinical improvement, whereas the perfusion deficits remained unchanged. These results further indicate that central serous chorioretinopathy is primarily a choroidal disease. PMID- 8420379 TI - Fluorescein angiography in the diagnosis of giant cell arteritis. AB - Clinical data and fundus fluorescein angiograms were analyzed from 35 patients with acute (onset less than four weeks) anterior ischemic optic neuropathy. Nineteen of the 35 patients (54%) had nonarteritic disease, and 16 patients (46%) had giant cell arteritis confirmed by biopsy. Patients with arteritis had higher erythrocyte sedimentation rates, larger cup/disk ratios, and delayed fluorescein dye appearance and choroidal filling times. Three additional patients with cranial arteritis confirmed by biopsy, but without visual loss, had angiographic characteristics similar to patients with arteritic ischemic neuropathy. We consider fluorescein angiography a valuable diagnostic adjunct in identifying patients with giant cell arteritis. PMID- 8420380 TI - Retinal vasoconstriction after laser treatment for diabetic macular edema. AB - The diameter of retinal arterioles, venules, and their macular branches was measured before and after macular laser photocoagulation in one eye each of six men and eight women with diabetic macular edema. The macular arteriolar branches constricted 20.2% (P < .001) and the venular branches constricted 13.8% (P < .001). This autoregulatory vasoconstriction results from the improved retinal oxygenation caused by the laser treatment. By extrapolating the principles of tissue edema formation in general, we hypothesized how macular laser treatment affects diabetic macular edema. Starling's law predicts that (laser-induced) vasoconstriction and reduced intravascular hydrostatic pressure should reduce edema formation in any tissue, including the retina. PMID- 8420381 TI - Early revision in the office for adults after unsatisfactory blepharoptosis correction. AB - The most common complications of levator palpebrae superioris muscle blepharoptosis repair are undercorrection, overcorrection, and abnormalities of the eyelid contour. Previously described nonsurgical as well as surgical methods delay the repair of such complications and introduce the same confounding factors that can affect judgment of the eyelid level as during the initial surgical procedure. Twenty-two patients underwent a highly predictable surgical technique to revise unsatisfactory postoperative eyelid positions. Twenty-five of 26 eyelids (96%) had a satisfactory result and only one of 26 (4%) remained undercorrected. The revision is performed three to four days after the initial blepharoptosis correction and involves blunt separation of the wound without local anesthesia. The levator aponeurosis is advanced or recessed and resutured to the tarsus to achieve the proper eyelid height and contour. The advantages of this revision technique are as follows: (1) the procedure can be quickly and easily performed in the office; (2) the anatomic defects are corrected; (3) sharp dissection, bleeding, and edema are avoided; (4) the technique is painless and usually requires no local anesthetic injections; (5) the tone and function of the levator palpebrae superioris muscle, Muller's, and orbicularis oculi muscles remain undisturbed intraoperatively; and (6) early correction is achieved, thereby enhancing patient acceptance. PMID- 8420382 TI - A biometric study of ocular changes during accommodation. AB - We performed a biometric study that used A-mode ultrasonography on 106 subjects during ocular accommodation. The subjects were divided into two groups; group 1 included 76 subjects and group 2 included 30 subjects. In group 1, we measured the anterior chamber depth, lens thickness, and axial length in the right eye while the left eye, wearing corrective spectacles, focused at distances of 6 m, 33 cm, and 33 cm with an additional correction of +3.0 diopters to offset any accommodative effect. In group 2, we measured the anterior chamber depth, lens thickness, and axial length in the right eye while the left eye focused at distances of 6 m, 33 cm, and 12.5 cm. Similar to the left eyes in group 1, the left eyes in group 2 wore corrective spectacles during all procedures. During accommodation, decreased anterior chamber depth and thickening of the lens were noted in all cases. In group 1, axial length significantly increased an average of 0.06 +/- 0.01 mm (P < .0005) while the left eye focused at a distance of 33 cm. There were no significant changes with the additional +3.0 diopters (P < .05). In group 2, axial length significantly increased an average of 0.05 +/- 0.01 mm (P < .0005) when the left eye focused at a distance of 33 cm, and there was further significant elongation of 0.05 +/- 0.01 mm when the left eye focused at a distance of 12.5 cm. Collectively, these results suggest that axial length increases along with changes in the lens and anterior chamber depth during ocular accommodation. PMID- 8420383 TI - Three-year follow-up of the Fluorouracil Filtering Surgery Study. AB - Patients who participated in the Fluorouracil Filtering Surgery Study, a clinical trial in which patients were randomly assigned to treatment to determine the efficacy and safety of subconjunctivally injected 5-fluorouracil after filtering surgery in eyes with poor prognoses, were followed up for three years. Treatments in 49 of the 100 eyes in the 5-fluorouracil group and 73 of the 99 eyes (74%) in the standard treatment group were classified as failures, defined by reoperation for control of intraocular pressure or intraocular pressure greater than 21 mm Hg during the first three years postoperatively (P < .001, chi-square). Late-onset leakage of aqueous through the filtering bleb occurred more frequently in the 5 fluorouracil group (seven of 105 eyes, 7%) than in the standard treatment group (none of 108 eyes, 0%) (P = .006, Fisher's exact test). We recommend the use of 5 fluorouracil after trabeculectomy in eyes after previous cataract extraction or unsuccessful filtering surgery. The increased risk of late-onset conjunctival filtering bleb leaks associated with 5-fluorouracil cautions against its routine use in patients with good prognoses. PMID- 8420384 TI - Corneal endothelial cell counts after Molteno implantation. AB - Implantation of a Molteno drainage shunt has been shown to be effective in advanced glaucoma. Foreign substances within the anterior chamber have been known to cause progressive endothelial cell loss. We undertook a study to evaluate the endothelial effects of an indwelling Molteno drainage shunt. Nineteen patients who underwent uneventful implantation of a Molteno drainage shunt for advanced aphakic or pseudophakic glaucoma were followed up. Serial endothelial cell counts were obtained in a masked fashion. During follow-up periods ranging from 5.4 to 25.7 months, endothelial cell loss averaged two cells per square millimeter per postoperative month with a 95% confidence interval of positive seven cells to negative ten cells per square millimeter per postoperative month. No clinically significant progressive trend in endothelial cell loss was seen in patients undergoing uncomplicated Molteno drainage procedures. Larger sample sizes with longer follow-up will be necessary to establish whether a Molteno drainage shunt causes clinically remarkable endothelial cell loss. PMID- 8420385 TI - Neuro-ophthalmologic findings in chordoma and chondrosarcoma of the skull base. AB - Review of the clinical features of 48 patients with chordoma and 49 patients with low-grade chondrosarcoma of the skull base disclosed overlapping clinical profiles but distinctive features. Both tumors occurred at all ages but chondrosarcoma tended to occur in the third and fourth decades. Twenty-five (52%) of the patients with chordoma and 24 (49%) of the patients with chondrosarcoma had ocular symptoms (diplopia or visual impairment) as the initial manifestation of the disease. Of the 59 patients (both groups) with diplopia, the diplopia was initially intermittent in 25 (42%). Headache and diplopia from an insidious abducens nerve palsy was most common in both groups. Abducens nerve palsy occurred in 22 (46%) of the patients with chordoma and 23 (47%) of the patients with chondrosarcoma. Normal examination results were more common in patients with chordoma, whereas visual loss, facial numbness, and multiple cranial neuropathies were more common in patients with chondrosarcoma. The similarities in the clinical features of these tumors reflect their common origin at the central skull base and the vulnerability of the abducens nerves at that site. The differences reflect the tendency of chordomas to originate from the clivus and chondrosarcomas to originate from the temporal bone. PMID- 8420386 TI - Tandem chromatographic-immunological analyses. AB - The function of conventional immuno-logical assays is to determine the presence and amount of known substances. Cross-reactivity of antibodies with unknown species of similar structure is a problem that can be circumvented by using chromatography in a second level of discrimination. Tandem LC-IA systems increase the selectivity and the sensitivity of assays by removing interfering species. For cases in which the goal is to prove that similar species are absent from a sample or to identify antigen variants, conventional assays fail. LC-IA systems are adept at dealing with these tasks. The necessary steps of metering reagents, separating Ag:Ab complex from other species, and enhancing detection in an immunological assay are easily accommodated by the LC system. In fact, the LC system provides a wide variety of new, high-sensitivity, high-speed methods for carrying out immunological assays. On the basis of the enormous versatility, selectivity, and sensitivity that LC-IA brings to analytical chemistry, we believe this technique will become increasingly important for monitoring analytes in complex biological matrices. PMID- 8420387 TI - Effects of buffer pH on electroosmotic flow control by an applied radial voltage for capillary zone electrophoresis. AB - Electroosmotic flow has been shown to be controlled via an applied radial voltage. Many factors determine the effectiveness of this control, and one major factor is buffer pH. In this study the effectiveness of the applied radial voltage for controlling electroosmotic flow while varying buffer pH is examined. Previously developed theory is applied and compared to experimental results for a pH range from 1.4 to 6.32. Analysis time is dramatically reduced by applying a radial voltage for separation of a peptide mixture at pH 1.4. Theory predicts laminar flow profiles under some conditions when applying this technique. However, experimental evidence at pH 6.32 and 1.4 shows no evidence of band broadening from a laminar flow profile. Theoretical and experimental results indicate the largest range of effective electroosmotic flow control via an applied radial voltage occurs at low ph. Furthermore, a sigmoidal relationship between electroosmotic flow and applied radial voltage is clearly apparent under these conditions. In contrast, at high buffer pH ( > 6) the relationship appears to be linear and is only over a limited range of flow velocities. PMID- 8420388 TI - Reduced surface area in mitotic rounding of human Chang liver cells. AB - The rounding up of mitotic human Chang liver cells in monolayer culture was studied quantitatively. It was surprising to find significant reduction in cell surface area considering that endocytosis has been demonstrated to be at a complete standstill in M phase. Uptake studies using impermeant BCECF (2',7' bis(2-carboxyethyl)-4(5)-carboxyfluorescein free acid) pH indicator and particulate neutral red dye in aqueous buffer showed preferential internalization into mitotic cells in direct contrast to expectation since interphase cells do not have arrested endocytosis. However, infolded plasma membrane ruffles and internalized extracellular material were demonstrated in prophase cells, much like those seen in interphase rounding via the induction of intracellular alkalinizations. Raised intracellular pH (pHi) is a universal and consistent finding in M phase cells. Despite cessation of small pit endocytosis, it remains possible for plasma membrane internalization to be a causal factor in the observed surface area reduction in mitotic rounding. PMID- 8420389 TI - Cryofixation of basement membranes followed by freeze substitution or freeze drying demonstrates that they are composed of a tridimensional network of irregular cords. AB - Since conventional chemical fixation may extract tissue components and thus alter structural organization, cryofixation was used to reexamine the ultrastructure of three thick basement membranes: lens capsule, Reichert's membrane, and Engelbreth Holm-Swarm (EHS) tumor matrix, and two thin basement membranes, those of epididymis and semi-niferous tubules. Cryofixation was achieved by slam freezing followed by either freeze substitution in dry acetone containing 1% osmium tetroxide and 0.05% uranyl acetate or freeze drying in a molecular distillation dryer. The results by both procedures demonstrate that thick basement membranes and the lamina densa of thin basement membranes are composed of a network of anastomosing strands referred to as cords. The cords vary in density and distinctiveness, but their thickness averages 3 to 5 nm in every tissue examined. The spaces separating the cords vary within wide limits, but their mean diameter is approximately 15 nm in every case. Two other common features are 1) the presence within the network of a few 1.5-3.0-nm-thick filaments and 2) 4.5-nm wide sets of parallel lines referred to as double tracks. When these results are compared with those previously described after conventional fixation, no significant difference is observed in either the cord network or the associated filaments and "double tracks." However, in the thin basement membranes processed by cryofixation, the lamina densa is in direct contact with epithelial cells, whereas, after conventional fixation, the lamina densa is separated from the epithelial cells by a pale layer referred to as lamina lucida or lamina rara. Immunogold labeling of three basement membranes after cryofixation and freeze substitution in acetone containing 0.3% glutaraldehyde yields strong reactions for laminin, type IV collagen, and heparan sulfate proteoglycan. Comparison with previous results indicates that conventional formaldehyde fixation adequately preserves laminin and type IV collagen but causes the loss of some proteoglycan. It is concluded that, except for this loss and the absence of lamina lucida in cryofixed thin basement membranes, the morphological and antigenic features obtained after cryofixation are similar to those observed in the past after conventional fixation. PMID- 8420390 TI - Possible nonlinear effects of exercise on bone in male subjects over age 60 years. AB - We investigated the relationships among bone mass, bone cell activities, and exercise level in 20 healthy 61-77 year old male volunteers divided into three groups according to the time they physically trained per week: nine subjects training less than 3 hr/week, five subjects between 3 and 6 hr/week and six subjects more than 6 hr/week. Physical performance was evaluated by VO2 max (ml min-1 kg-1). After tetracycline double labeling, iliac crest biopsy was obtained from each subject. The longer the physical activity, the higher the VO2 max. Subjects exercising between 3 and 6 hr/week revealed higher adjusted appositional and bone formation rates than all the others; mass and structural parameters also showed higher (nonsignificant) values. For the whole population VO2 max appeared negatively related to cortical thickness, cancellous bone volume, and trabecular thickness. These alterations were accompanied by increased cancellous bone turnover; this was evidenced by an increase in activation frequency and in resorption and formation rates as VO2 max increased. The bone remodeling periods tended to decrease also. Whatever the bone turnover rate, subjects were in steady state as far as their bone balance was concerned. Relationships between VO2 max and mineral apposition rate on the one hand and VO2 max and resorption surface on the other hand were best fitted by a quadratic model, suggesting a possible nonlinear effect of physical training on bone mass. We hypothesize that there is a threshold (6 hr/week) determining different effects. Adjustment of bone mass and trabecular arrangement were completed at time of biopsy and reflected probably past and transient bone imbalance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420391 TI - Bone growth and periosteal migration control masseter muscle orientation in pigs (Sus scrofa). AB - During growth the muscles of mastication alter their lines of action. Research on long bones indicates that the apparent migration of muscle attachments is due to the movement of the periosteum relative to the underlying bone. To assess whether the pig masseter muscle follows the periosteum during growth, implants of titanium granules in a gelatin matrix were placed simultaneously in various parts of the masseter muscle and its periosteal and bony attachments. Growth movements of these tissues were followed radiographically for 2 months. Granule position was verified histologically. Periosteal movement was the dominant growth process at the insertion of the masseter. All implants migrated caudally relative to the mandible. However, a strong position effect was seen dorsoventrally: implants placed high in the ascending ramus migrated dorsally as well as caudally; low implants migrated only caudally. This differential migration, ascribed to the influence of the condyle, accounts for the increasing horizontal orientation of dorsal fibers. A similar differential was seen along the rostrocaudal axis of the ramus. In contrast to the insertion, the origin of the masseter from the zygomatic arch shows no periosteal movement. Rather, the entire bone-muscle complex becomes displaced by sutural growth, leading to increasing vertical orientation of the masseter. Thus two different aspects of skull growth are responsible for the change in muscle anatomy. PMID- 8420392 TI - Morphogenesis in the fetal rat proximal colon: effects of cytochalasin D. AB - Two major morphogenetic events, epithelial conversion and fold formation, occur in the proximal rat colon during the last week of gestation. To evaluate the role of actin microfilaments in these two developmental processes, explants from the proximal colon of 19 day fetal rats were cultured in the presence of vehicle (0.1% dimethylsulfoxide), 0.1, 1.0, or 10 micrograms/ml of cytochalasin D (CD) for 24-48 hr. Explants as well as 19, 20, and 21 day in vivo controls were prepared for light, fluorescence, and electron microscopy. The distribution of actin filaments was determined by rhodamine-conjugated phalloidin binding and ultrastructural analysis of tissue fixed in the presence of tannic acid. Prior to fold formation, phalloidin binding was enhanced along the entire epithelial mesenchymal interface. At the onset of fold formation, focal areas of intense fluorescence appeared at irregular intervals along the base of the stratified epithelium. Within 1 day, these focal intensities were localized at the apex of small forming folds. Additional changes occurring at the epithelial-mesenchymal interface in association with fold formation included: 1) ruffling of the previously smooth basal lamina, 2) a shape change in the subjacent mesenchymal cells from elongate to cuboidal along with the appearance of numerous processes abutting the basal lamina, and 3) a unique orientation of the associated collagen fibrils in some presumptive folds. Fold formation was inhibited in > 93% of explants cultured in the presence of 1.0 micrograms/ml CD. These explants appeared to be arrested precisely at the onset of fold formation. Epithelial conversion was also incomplete in these explants. These findings indicate an active role for actin in both fold formation and epithelial conversion. PMID- 8420393 TI - Vasocontractions of the in-vitro toad aortas induced by endothelin-1 and sarafotoxin-S6b. AB - Dose-dependent tension curves were recorded from the in vitro toad aortas by administration of endothelin-1 and sarafotoxin-S6b. The maximal contractile tensions by both drugs were evoked at a 10(-8) M concentration. By a single dose application (10(-8) M) of endothelin-1 and sarafotoxin-S6b to both endothelium preserved and denuded vessels, the induction of the endothelium-dependent vasocontraction occurs after 2 min of administration. Ultrastructural changes of Weibel-Palade bodies such as decrease in electron density, swelling with a wide peripheral halo, and expulsion of their contents in a manner of exocytosis become evident within 2 min after administration of these drugs. These findings indicate that some vasocontractile substances in Weibel-Palade bodies are extracellularly discharged by endothelin-1 and sarafotoxin-S6b. PMID- 8420394 TI - Early formation of the vascular system in quail embryos. AB - The relation between vascular development and translocation of the splanchnic mesodermal layers was studied in presomite to 20-somite quail embryos by scanning electron microscopy. In addition, serially sectioned embryos were stained immunohistochemically with monoclonal antibodies (alpha QH1 or alpha MB1) specific for endothelial and hemopoietic cells. By the formation of the foregut the anterior borders of the two splanchnic mesodermal layers of a presomite embryo are translocated to the lateral and ventral sides of the foregut and fuse in the ventral midline of a 4-somite embryo. Meanwhile the splanchnic mesoderm differentiates into a splanchnic mesothelial layer and a plexus of endothelial cells, facing the endoderm. From 4 somites onward the foregut is covered by a single endothelial plexus. At first the endothelial precursors bordering the anterior intestinal portal and those in the area of the ventral mesocardium lumenize, subsequently giving rise to the endocardium of the heart tube. Hereafter, the pharyngeal arch arteries and the dorsal aortae develop from the remaining precursors. During formation of the pharyngeal arches, the pharyngeal arch arteries maintain their connections with the splanchnic plexus through the developing ventral pharyngeal veins. After disappearance of the dorsal mesocardium, the midpharyngeal endothelial strand, which is a longitudinal strand of proendocardial cells, remains connected to the foregut. This strand will contribute to the formation of the pulmonary venous drainage into the left atrium. A bilateral accumulation of cardiac jelly developing between the promyocardium and proendocardial plexus only suggests that the heart develops from two tubes. The proendocardial layer, however, is not divided by the ventral mesocardium but initially forms just one endocardial heart tube. PMID- 8420395 TI - Effects of prolactin on alpha and beta chloride cells in the gill epithelium of the saltwater adapted tilapia "Oreochromis niloticus". AB - Tilapia (Oreochromis niloticus), 21 g average body weight, were divided into two groups. A group was maintained in fresh water, whereas another group was adapted for 2 weeks to 20% salt water. Among the latter, fishes were injected every 2 days for a week with tilapia prolactin (ti-PRL I). Gills were prepared for electron microscopy in order to determine the types and surface areas of chloride cells in each experimental condition. Two types of chloride cells, the alpha and beta cells were easily distinguished on the basis of their location and ultrastructural features in the gills of freshwater fishes, while only one type of cell, the saltwater alpha cells presumably derived from the transformation of the freshwater alpha cells, were encountered in saltwater adapted animals. After PRL injection of saltwater adapted fishes, small chloride cells, which displayed ultrastructural features similar to those of beta cells in freshwater tilapia, reappeared in interlamellar regions of the gills. In the same experimental conditions, the voluminous saltwater alpha cells showed a tendency to resume ultrastructural features more characteristic of the freshwater alpha cells from which they were derived. These observations tend to indicate that prolactin behaves as a "freshwater adapting hormone" and that beta cells are specifically involved in fish adaptation to freshwater living conditions. PMID- 8420396 TI - Dendritic cell/lymphocyte clustering: morphologic analysis by transmission electron microscopy and distribution of gold-labeled MHC class II antigens by high-resolution scanning electron microscopy. AB - Dendritic cells (DCs) are potent antigen-presenting cells for a variety of immune responses; however, their mechanism of action has not been established. It is known that DCs can cluster with one another and with other cell types during in vitro immune responses, and clustering may be essential for the activation of resting lymphocytes. In this study, ultrastructural examination of clusters that form during extended culture of enriched rat splenic DCs (approximately 70% DCs) is reported. DCs were readily distinguished from other cell types, which included lymphocytes and macrophages. DCs displayed characteristic veils and/or dendritic processes that intertwined with processes of other cells within the cluster, or extended from the cluster periphery. Occasional DCs contained large vacuoles lined with small vesicles. A paramount feature of DCs is their constitutive expression of high levels of surface major histocompatibility complex class II antigens. The surface distribution of class II antigens on clustering DCs was examined using 10 nm immunogold labeling techniques and high-resolution scanning electron microscopy. DCs were readily distinguished by morphologic criteria, and examination of various surface membrane regions revealed a differential distribution of class II antigens. Gold label was frequently distributed in linear arrays and clusters, suggesting a cytoskeletal role in the recycling/redistribution of Class II antigens. These morphologic findings further an understanding of basic DC biology and their mechanism of action as antigen presenting cells. PMID- 8420397 TI - A study of the chick thymus microenvironment during development: analysis by monoclonal antibodies against thymic epithelium. AB - The process of T-lymphocyte differentiation within the thymus involves a series of molecular interactions. In this work we have carried out an analysis of the chick thymus microenvironment in order to evaluate its heterogeneity during development. We have produced 11 monoclonal antibodies whose staining patterns detected by the immunoperoxidase technique allowed us to divide them into five groups. A first group (E19-E2, P0-E5, and P15-T1) binds to thymic medullary stroma showing a reticular pattern on medullary epithelial cells and whose significance would be related to thymic stromal secretion. The second group of monoclonal antibodies (P15-T3) stains thymic corpuscles of 10- and 15-day chicks. The third group of antibodies includes P0-E1, P0-E3, P5-A6, and P15-T2 whose staining pattern is both medullary and cortical. The fourth group (P10-HB1 and P10-HB2) binds to thymic stromal and cortical thymocytes, and the fifth group (P5 A1) is characterized by the staining of medullary vessels of 5-day chicks. These five groups of monoclonal antibodies corroborate the existence of an antigenic diversity of the chick thymus microenvironment. Their possible relationships with T-cell differentiation and stromal-thymocyte interactions are discussed. PMID- 8420398 TI - Proliferative and structural differences between male and female mouse submandibular glands. AB - Sexual dimorphism has been observed in salivary glands of many species. In this study, evidence for sexual differences in adult mouse submandibular gland is extended beyond parenchymal cell composition, size, and volumes to include patterns of DNA synthesis and complexity of ductal branching. Computer-assisted three-dimensional reconstructions also revealed differences in overall organization of secretory complexes. Consistent with observations by others, granular intercalated duct cells were absent, while striated granular duct cells were low in proportion in the male glands relative to female glands. When the mean of average cell volumes were compared, acinar (AC) cells were smaller than granular duct (GD) cells in the male, but in the female the reverse was true. Furthermore, in addition to differences in average volumes of GD cells, the average volume of AC cells was significantly greater in females than males. The most dramatic evidence for sexual dimorphism was observed following a 90-min labeling with 3H-thymidine. Though all cell types showed DNA replication activity, the intercalated duct (ID) cells were substantially more active than AC and GD cells in the female, while in the male the GD cells, ID cells, and AC cells all showed approximately equal activity. Three-dimensional reconstructions indicated that the female possessed a more highly branched intercalated duct system and that the GD usually terminated within a secretory complex, whereas in males the GD typically passes through a secretory complex and forms a prominent cap-like structure on the opposite side. PMID- 8420399 TI - Morphometric changes in growth of the rat pelvis after papain administration. AB - The proteolytic enzyme, papain, was given systematically to evaluate the short- and long-term effects of inhibition of endochondral bone formation on pelvic growth, with emphasis on the innominate bone. Ninety-eight Lewis-strain male rats, used concurrently for craniofacial growth studies, were divided into two groups. Thirty rats from Group I (n = 48) received 2% crude papain i.p. daily from 25-40 days-of-age and were euthanized at 40, 54, and 70 days-of-age. Thirty five rats from Group II (n = 50) were given papain at the same dose from 25-70 days-of-age and were euthanized at 26, 40, 54, 70, and 120 days. The remaining animals in both groups were the controls. Standardized dorsoventral pelvic radiographs were taken of all 98 animals. Ten linear dimensions were measured on each and the data evaluated statistically. A reduction both in size and rate of growth of the bony pelvis was found. All the anteroposterior and most of the transverse pelvic dimensions were significantly shorter, to a greater extent in the prolonged papain group. Bi-ischial width was increased, perhaps to compensate for pelvic shortening and to accommodate the pelvic contents. The findings may contribute to our better understanding of abnormal endochondral bone growth in the pelvis. PMID- 8420400 TI - Developmental differences in the ossification process of the human corpus and ramus mandibulae. AB - A correlation was sought between the organization of the dental crest and the ossification of the corpus mandibulae in 14 human embryos and 13 human fetuses. The different types of ossification between the corpus and the ramus mandibulae suggest that the cartilago mandibularis (meckeliensis) guides the formation of the mandibula, while the dental crest acts as a coorganizer. In the area of the foramen mentale, the lamina dentalis begins to invaginate (to give rise to the dental crest), and at this level intramembranous ossification of the corpus mandibulae commences. These findings, together with the presence of the cartilago mandibularis before the appearance of the dental crest, and the fact that the former is seen along the entire length of the mandibula (from the symphysis mandibulae to the capsula otica), support the hypothesis that the dental crest, rather than the cartilago mandibularis, acts as the coorganizer in the corpus mandibulae. PMID- 8420401 TI - Star volume in bone research. A histomorphometric analysis of trabecular bone structure using vertical sections. AB - Conventional bone histomorphometry performed on iliac crest biopsies does not generally provide unbiased stereological estimates of parameters related to bone structure due to the anisotropy of trabecular bone; this, however, can be obtained with vertical sections, which are anisotropic sections, in combination with an anisotropic test system. A practical procedure for obtaining vertical sections from bone is described. The new stereological parameter, the star volume, can provide an unbiased estimation of the absolute mean size of the marrow space and thus give an indirect estimate of the connectivity of trabecular bone structure. The marrow space star volume of vertebral bodies and iliac crest increases with age in both sexes demonstrating that the structural bone changes which occur with age is a topological one with changes in trabecular connectivity. The practical procedure including sampling efficiency for marrow space star volume is described. PMID- 8420402 TI - Leukotrienes in bile during the early and the late airway responses after allergen challenge of sensitized rats. AB - The Brown Norway rat produces high levels of IgE in response to active immunization and develops both early and late airway constrictor responses after subsequent allergen challenge. We have used this model of allergic asthma to investigate the temporal relationship between the in vivo synthesis of peptidoleukotrienes (peptido-LTs) and the late response. Brown Norway rats that had been sensitized by injection of ovalbumin 2 to 3 wk prior to the commencement of the experiment were subjected to bile duct cannulation and tracheal intubation. The rats were challenged 2 h later by intratracheal instillation of ovalbumin. Lung resistance was measured before and at frequent intervals after antigen challenge. Biliary peptido-LTs (LTC4, LTD4, LTE4, and N-acetyl-LTE4) were measured by a combination of high pressure liquid chromatography and radioimmunoassay in bile samples collected for a period of 1 h before instillation of ovalbumin, and between zero and 1 h, 1 and 4 h, 4 and 6 h, and 6 and 8 h, subsequently. All of the 10 rats subjected to antigen challenge developed early responses. Of these, six also developed late responses, whereas two died about 1 h after challenge. The levels of peptido-LTs excreted in bile between 4 and 8 h after antigen challenge (corresponding in time to the late responses) were about four times higher in the ovalbumin-instilled rats that developed late responses (n = 6) than in the ovalbumin-sensitized control rats that had been subjected to instillation of saline (n = 6; p < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420403 TI - Acute effects of ambient ozone on pulmonary function of children in The Netherlands. AB - In the spring and summer of 1989 an epidemiologic study was conducted to evaluate the acute effects of photochemical air pollution episodes on pulmonary function of children living in three nonindustrial towns in the Netherlands. Spirometry was performed repeatedly in the schools of the children, mostly during the morning hours. Data from 533 children having more than four valid pulmonary function tests were included in the analyses. The association between previous day ambient ozone concentration and pulmonary function was evaluated, using individual linear regression analysis and subsequent evaluation of the distribution of individual regression coefficients. One hour maximum ambient ozone concentrations frequently exceeded 160 micrograms/m3 but were all lower than the Dutch Air Quality Guideline of 240 micrograms/m3 for all three populations. Significant negative associations of previous-day ambient ozone with FVC, FEV1, peak expiratory flow (PEF), and maximal midexpiratory flow (MMEF) were observed. There were indications of systematic differences in responses among the children. Children with chronic respiratory symptoms did not have a stronger response than children without these symptoms. PMID- 8420404 TI - Effect of ambient winter air pollution on respiratory health of children with chronic respiratory symptoms. AB - The acute respiratory effects of ambient air pollution were studied in a panel of 73 children with chronic respiratory symptoms in the winter of 1990 to 1991. The participating children were selected from all children aged 6 to 12 yr in Wageningen and Bennekom, two small, nonindustrial towns in the east of the Netherlands. Peak flow was measured twice daily with MiniWright meters. A diary was used to register the occurrence of acute respiratory symptoms and medication use by the children. Exposure to air pollution was characterized by the ambient concentrations of sulfur dioxide (SO2), nitrogen dioxide (NO2), black smoke (BS), and particulate matter less than 10 microns (PM10). Associations between air pollution concentrations and health outcomes were analyzed using time series analysis. During the study period an air pollution episode occurred, with moderately elevated concentrations of PM10 and SO2. There were 6 days with 24-h average PM10 concentrations in excess of the WHO suggested lowest observed effect level of 110 micrograms/m3. After adjustment for ambient temperature, there were small but statistically significant negative associations of PM10, BS, and SO2 with both morning and evening PEF. There was a consistent positive association between PM10, BS, and SO2 with the prevalence of wheeze and bronchodilator use. Overall, the observed associations suggest a mild to moderate response to these moderately elevated levels of air pollution in a group of potentially sensitive children. PMID- 8420405 TI - Spirometry reference values for women and men 65 to 85 years of age. Cardiovascular health study. AB - Pulmonary function was assessed by spirometry in 5,201 ambulatory elderly participants of the Cardiovascular Health Study, sampled from four communities. A stringent quality assurance program exceeded American Thoracic Society (ATS) recommendations for spirometry. Less than 6% of the participants were unable to perform three acceptable spirometry maneuvers. A "healthy" subgroup of 777 women and men 65 to 85 yr of age was identified by excluding smokers and those with lung disease and other factors determined to independently, significantly, and negatively influence the FEV1. Results from black participants were examined separately. Reference equations and normal ranges for FEV1, FVC, and the FEV1/FVC ratio were determined from the healthy group. The results demonstrate differences in predicted values as great as 20% (0.5 to 1 L) for elderly patients when compared with the spirometry reference equations that are most commonly used in the United States. PMID- 8420406 TI - Cytokines in metal fume fever. AB - Metal fume fever is a flulike illness caused by zinc oxide inhalation and accompanied by an impressive pulmonary cellular response. We hypothesized that the syndrome is mediated by cytokines released in the lung after exposure to zinc oxide fume. We carried out 26 experimental welding exposures in 23 volunteer subjects, performing postexposure bronchoalveolar lavage (BAL) 3 h (n = 6), 8 h (n = 11), or 22 h (n = 9) after exposure. We detected tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-8 (IL-8) varying in a time- and exposure-related manner. The concentration of TNF in the BAL fluid supernatant was significantly greater at 3 h than at 8 h or 22 h after exposure (p < 0.05), exhibiting a statistically significant exposure-response relationship to airborne zinc at each follow-up time period (p < 0.05). TNF concentrations were statistically correlated with those of IL-6 in BAL supernatant obtained at 22 h (r = 0.78, p = 0.01) and with concentrations of IL-8 in BAL 8 h after exposure (r = 0.85, p = 0.001). IL-6 displayed a significant exposure-response relationship to zinc (p < 0.05) at 22 h. IL-8 exhibited a significant exposure-response relationship to zinc (p < 0.05) at 8 h after exposure, a time at which IL-8 correlated with marked increases in BAL fluid polymorphonuclear leukocytes (PMN) (r = 0.7, p = 0.01). Although we also detected interleukin-1 (IL-1) in BAL samples, this cytokine did not demonstrate a statistically significant exposure response. TNF, IL-6, and IL-8 in BAL fluid supernatant concentrations increased in a time and exposure-dependent fashion after zinc oxide welding fume exposure. The time course of increased cytokines, their correlations with one another and with PMN in the BAL fluid, and the consistency of our findings with the known kinetics and actions of these cytokines support the hypothesis that a network of cytokines is involved in the pathogenesis of metal fume fever. PMID- 8420407 TI - ELISA detection of IL-1 beta in human sera needs independent confirmation. False positives in hospitalized patients. AB - The detection of cytokines in human sera has become an accepted index of disease activity in various diseases, including sepsis. However, little attention has been paid to the specificity of these measurements. Using a sensitive sandwich enzyme-linked immunoassay (ELISA), we studied IL-1 beta detectability in sera from 419 serum samples randomly obtained from our clinical laboratory. In initial studies, 6.7% of samples were positive (n = 24, 0.1 to 1.0 ng/ml, and n = 5, 1 to 80 ng/ml). However, attempts to further characterize positive samples revealed that over 90% were falsely positive. For example, samples fractionated on Sephadex G-75 demonstrated IL-1 beta "detectability" near the void volume, and negative samples, spiked with rIL-1 beta, eluted at approximately 17 kD. To determine if this detectability was due to heterophillike antibodies, 23 of 29 "positive" samples were retested in the presence of nonimmune mouse serum. Only 2 of 23 previously positive samples were still positive. Importantly, mouse serum had no effect upon normal human serum spiked with rIL-1 beta. Furthermore, blinded samples sent to a reference laboratory also demonstrated false positive IL-1 beta detection in selected samples. Taken together, these data demonstrate that the presence of nonspecific immunoactivity in sera may confound cytokine assays of human biologic material and suggest that, when possible, a second means of confirming ELISA-positive samples be used. PMID- 8420408 TI - A general mathematical model for respiratory dynamics relevant to the clinical setting. AB - We have developed and validated a general mathematical model for the dynamic behavior of the single-compartment respiratory system in response to an arbitrary waveform of applied inspiratory pressure. Our general model for ventilation applies to all integrable functions of applied pressure, and it enables computation of most ventilation and pressure variables of clinical interest from clinician-selected and impedance-determined inputs readily measured or estimated at the bedside. Interactions between both phases of the ventilatory cycle are considered by assuming that deflation occurs passively from a unicompartment lung. Because this flexible model appears both capable of accurate prediction and robust to major violations of its underlying linear assumptions, it may prove to be of value in a variety of scientific, educational, and clinical settings. PMID- 8420409 TI - Bronchoalveolar lavage. Quantitation of intraalveolar fluid? AB - A precise calculation of the amount of intraalveolar fluid is the basis of a quantitative analysis of intraalveolar compounds. Different approaches have been made to cover this important problem. Here, we report a comparative study with five markers: 99mTc-DTPA, 51Cr-EDTA, inulin, urea, and methylene blue in animal experiments as well as in human experiments. The marker substances were added to the lavage fluid, and the "dilution" of the markers, i.e., the alveolar fluid, was calculated. The results showed that in animals with healthy lungs the tracer methods are able to calculate amounts of intraalveolar fluid that are comparable to morphologic findings. In animals as well as in humans, methylene blue and inulin were shown to be useless in determining alveolar fluid volume compared with the tracer methods. In humans, the calculations with the urea method and with Tc-DTPA were in the same magnitude, but there was no individual correlation. We conclude that, at present, the methods to quantitate alveolar fluid volume lack precision and add nothing to a deeper understanding of alveolar biology. PMID- 8420410 TI - A mechanism of erythromycin treatment in patients with diffuse panbronchiolitis. AB - Recently, "low-dose and long-term" erythromycin treatment has been reported as effective on diffuse panbronchiolitis (DPB), but its mechanism is still obscure. Patients with DPB were found to have significantly higher percentages of neutrophils in the pre-erythromycin treatment bronchoalveolar lavage fluid (BALF) than healthy nonsmoking volunteers (p < 0.001). They showed a significant reduction in BALF neutrophil percentages after erythromycin treatment (p < 0.01). The neutrophil chemotactic activity (NCA) was significantly elevated in BALF obtained from 19 patients with DPB compared with that from healthy volunteers (p < 0.001). A significant reduction in the NCA was observed in post-erythromycin treatment BALF of 11 patients with DPB (p < 0.001). Additionally, there was a significant correlation between the reduction of NCA and neutrophil percentage in pre- and post-erythromycin treatment BALF (r = 0.726, p < 0.05). Finally, we investigated the effect of erythromycin on the intrapulmonary influx of neutrophils by intratracheal injection of lipopolysaccharide (LPS) and interleukin-8 (IL-8) in mice. The intrapulmonary influx of neutrophils was significantly suppressed (p < 0.001) in mice intraperitoneally injected with erythromycin at 5 mg per animal 2 h before intratracheal injection of LPS (control group: 6.5 +/- 1.6 x 10(5) versus erythromycin-treated group: 1.7 +/- 0.5 x 10(5)), but not 10 h before lung challenge. This inhibition was observed at 6 h after lung challenge and became maximal with 84% suppression at 24 h. Week long administration of erythromycin did not alter the intrapulmonary influx of neutrophils. The number of neutrophils in the peripheral blood was not affected by erythromycin, indicating that the drug was not toxic.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420411 TI - Heparin inhibits the immediate response to antigen in the skin and lungs of allergic subjects. AB - Although heparin is used as an anticoagulant, its biologic function remains unclear. Substantial evidence exists that suggests it may modulate many aspects of immune function and inflammation. We demonstrated, in a double-blind, placebo controlled, crossover study involving 10 allergic subjects, that a small dose of heparin (25 U/kg) administered intravenously 10 min before challenge reduced the acute cutaneous reaction to 10 allergens and histamine from a group-average sum of mean (+/- SD) wheal diameters at a baseline of 29.9 +/- 10 mm and after normal saline placebo (29.5 +/- 10.7 mm) to after heparin (14.4 +/- 10.4 mm) (p < 0.02, Wilcoxon's signed rank test). In 15 subjects with asthma and dust mite allergy, nebulized heparin 20,000 units administered in a double-blind, placebo controlled, crossover fashion 10 min before challenge inhibited the bronchospasm induced by inhaled dust mite extract. Log2 of the provocative dose of mite extract causing a 20% fall in FEV1 at baseline was 4.1 +/- 1.5 protein nitrogen units (PNU); after normal saline it was 4.5 +/- 2.0 PNU, and after heparin it was 5.1 +/- 2.5 PNU (p = 0.04). These data suggest heparin may have an inhibitory role in acute mast-cell-mediated allergic inflammation. PMID- 8420412 TI - Seals, seal trainers, and mycobacterial infection. AB - In 1986, three seals died in a marine park in Western Australia; culture of postmortem tissue suggested infection with Mycobacterium bovis. In 1988, a seal trainer who had been employed at the Western Australian marine park until 1985 developed pulmonary tuberculosis caused by M. bovis while working in a zoo 3,000 km away on the east coast of Australia. Culture characteristics, biochemical behavior, sodium dodecyl sulphate polyacrylamide gel electrophoresis, and restriction endonuclease analysis suggested that the strains of M. bovis infecting the seals and trainer were identical but unique and differed from reference strains and local cattle strains of M. bovis. The infection in both the seals and the trainer had a destructive but indolent course. This is the first time that M. bovis has been observed in seals and the first time that tuberculous infection has been documented to be transmitted from seals to humans. Further investigation of the extent of tuberculous infection in seal populations elsewhere in the world seems warranted, and those working with seals and other marine animals should be monitored for infection. PMID- 8420413 TI - Neutrophil sequestration and migration in localized pulmonary inflammation. Capillary localization and migration across the interalveolar septum. AB - Recent studies have raised the possibility that biophysical properties of the neutrophil, specifically cell size and deformability, along with the unique hemodynamic properties of the pulmonary microvasculature may be important determinants of neutrophil transit through the lung and lead to significant retention in capillaries. Additionally, accumulating evidence supports the concept that leukocytes may emigrate into inflammatory lesions through the pulmonary capillary wall, quite unlike the situation in the systemic circulation where the predominant site of margination and emigration is the postcapillary venule. To confirm and extent this suggestion, morphometric techniques were employed to determine the location and kinetics of neutrophil movement from the vascular compartment to the alveolar space in response to an inflammatory reaction initiated by local instillation of fragments of the activated form of the fifth component of complement (C5f). We found that the site of neutrophil sequestration and probable migration was almost exclusively the capillary. Neutrophil sequestration appeared first in the capillaries of the interalveolar septae, with negligible accumulation in arterioles and venules (less than that in interalveolar septae by a factor of 10(4) when corrected for the relevant surface areas). Accumulation in the airspaces lagged behind that in the interalveolar septa by about 60 min, placing an upper time-limit on the emigration process across the epithelial barriers. Interestingly, neutrophils samples by bronchoalveolar lavage represented only 1 to 2% of those neutrophils shown to be present in the airspaces of the lung by morphometric assessment, suggesting that bronchoalveolar lavage may sample only a subpopulation of the emigrated cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420414 TI - Induction of heat stress proteins is associated with decreased mortality in an animal model of acute lung injury. AB - This study examined the hypothesis that transient, whole-body hyperthermia would reduce lung damage and/or mortality in a previously described animal model of acute lung injury. Normal, adult Sprague-Dawley rats were randomly assigned either to a heated (n = 40) or to a sham-heated (n = 49) group. Heated animals were warmed to 41 to 42 degrees C 18 h before intratracheal instillation of phospholipase A2. Forty-eight hours after phospholipase A2 exposure, the two groups were compared in a blinded fashion for mortality rate, PaO2, AaPO2, lung wet/dry weight ratio, alveolar inflammatory cell number, and lung histopathology. Heated, injured animals exhibited a reduced mortality rate and less lung damage than did unheated animals: mortality (zero versus 27%, p < 0.001); AaPO2 (22 +/- 3 versus 36 +/- 15 mm Hg, p < 0.002); lung lavage cell counts (5.3 +/- 3 versus 16.9 +/- 7 x 10(6)/ml, p < 0.05); lung wet/dry weight ratio (4.1 +/- 0.6 versus 5.1 +/- 0.7, p < 0.025); parenchymal lung injury fraction (0.10 versus 0.51, p < 0.001). Transcription and translation of heat shock proteins (HSP70) were examined by Northern and Western analysis. Pulmonary tissue HSP70 mRNA was elevated 1 h after heating. HSP72 protein levels were increased over baseline levels between 12 and 72 h after whole-body hyperthermia, but they were unchanged in sham-heated animals. These data indicate that thermal pretreatment associated with the induction of HSP72 protein synthesis, attenuates tissue damage and mortality in experimental lung injury. PMID- 8420415 TI - Site of pharyngeal narrowing predicts outcome of surgery for obstructive sleep apnea. AB - Uvulopalatopharyngoplasty (UPPP), an operation that enlarges the pharyngeal airway at the level of the soft palate, improves respiratory status during sleep in only 50% of patients with obstructive sleep apnea (OSA). This poor outcome suggests that narrowing of the pharyngeal airway at nonpalatal sites contributes to the obstructive process in many patients with OSA. We have used a novel endoscopic method to identify regions of the passive pharyngeal airway most susceptible to narrowing or complete closure. In order to test the hypothesis that narrowing of the passive airway at the nasopharynx predicts a favorable surgical outcome, we have preoperatively assessed the local mechanics of the passive pharyngeal airway in 18 patients with OSA undergoing UPPP. The patient population was prospectively divided into two groups: an exclusively nasopharyngeal (ENP) group, consisting of patients exhibiting narrowing only in the nasopharynx, and a not exclusively nasopharyngeal (NENP) group, consisting of patients having at least one site of narrowing outside the nasopharynx. The frequency of respiratory disturbances and arousals and the cumulative time in apnea-hypopnea were significantly reduced after surgery for the ENP group, but not for the NENP group. Improvement rate for the ENP group (86%) exceeded that for the NENP group (18%) (p < 0.01). These differences became even greater when selection criteria for the ENP group were made more restrictive (i.e., restricted to the velopharynx) or more liberal (i.e., including secondary narrowing of the oropharynx). Our results show that evaluation of passive pharyngeal mechanics identifies patients with OSA likely to improve after UPPP. PMID- 8420416 TI - Role of chemical drive in recruiting upper airway and inspiratory intercostal muscles in patients with obstructive sleep apnea. AB - Upper airway dilating muscle activity increases during apneic episodes in patients with obstructive sleep apnea (OSA). To elucidate the relative contribution of chemical and nonchemical stimuli to augmentation of the upper airway dilating muscle, we measured the response of genioglossus muscle (GG) and inspiratory intercostal muscle (IIM) activities to obstructive apnea during non REM sleep and compared them with the response to progressive hypoxia and hypercapnia during awake periods in seven male patients with OSA. GG EMG was measured with a wire electrode inserted percutaneously, and IIM EMG was measured with surface electrodes placed in the second intercostal space parasternally. Responses to hypoxia and to hypercapnia were assessed by rebreathing methods in the supine position while awake. Following these measurements, a sleep study was conducted with the EMG electrodes placed in the same locations. The relationship between GG and IIM activities during the cycle of apnea and postapneic ventilation in non-REM sleep was quasi-linear, and the slope of the regression line was significantly greater than those during progressive hypoxia and progressive hypercapnia. The amplitude of GG activity at 70% of maximum IIM activities in the hypoxic test was 140 +/- 20% (mean +/- SEM) during non-REM sleep, which was also significantly greater than that during hypoxia (51 +/- 10%) and that during hypercapnia (59 +/- 15%). These results suggest that nonchemical factors contribute considerably to augmentation of GG activity during obstructive apneic episodes. The nonchemical stimuli may arise from mechanoreceptors activated by upper airway obstruction and behavioral factors associated with change in sleep states. PMID- 8420418 TI - Chronic beryllium disease in a dental laboratory technician. AB - Workers involved in the manufacture of dental prostheses are exposed to a number of potentially harmful substances capable of inducing lung disease. In this report, we describe a dental laboratory technician who developed chronic beryllium disease as a result of exposure in the workplace. The diagnosis of chronic beryllium disease was suspected from the clinical, radiographic, and histologic features and confirmed by the in vitro proliferation of lung lymphocytes to beryllium salts. The potential risks of beryllium use in the dental industry have been recognized for some time, but this is the first documentation of chronic beryllium disease in this population of workers. Since chronic beryllium disease may be easily confused with sarcoidosis, awareness of this occupational association is essential in preventing misdiagnosis and in providing appropriate management. PMID- 8420417 TI - Klippel-Feil sequence and sleep-disordered breathing in two children. AB - We report two children with severe sleep-disordered breathing associated with Klippel-Feil sequence. In both patients, minor vertebral anomalies were associated with a major hindbrain anomaly. In one child, the Klippel-Feil sequence had been diagnosed previously, but the hindbrain anomaly was not recognized. Two years later, this child developed fatal obstructive sleep apnea. In the other child, neither the Klippel-Feil sequence nor hindbrain anomaly had been identified before the child's presentation with sleep-disordered breathing characterized by bradypnea and stridor. Because many of the complications of hindbrain anomalies may be amenable to neurosurgical treatment, we recommend that patients with Klippel-Feil sequence be followed for the development of sleep disordered breathing. Sleep complaints need prompt evaluation with polysomnography, whereas neurologic signs require imaging with attention to the cervicomedullary junction. Unsuspected CNS disorders must be considered in children who present with stridor or serious respiratory disturbances during sleep. PMID- 8420419 TI - A bronchofibroscopic analysis of the bronchial mucosa using a dye-scattering method for the detection of bronchial lesions. AB - A study was undertaken to evaluate the usefulness of a dye-scattering method for the analysis of bronchial lesions. After methylene blue was introduced through a cannula, the bronchial staining intensities seen by bronchofibroscopy were later compared with the histologic findings in 74 biopsied specimens of lesions taken from 20 patients with lung cancer and from 6 patients with bronchiectasis, 2 with a rheumatoid lung, and 2 with sarcoidosis. It was found that 86% (25 of 29) of the specimens that stained positively were malignant. Further, 4 specimens that manifested a false positive response to staining were either too small to be inspected histologically or showed necrosis. Of 27 specimens that were negative to staining, 89% (24 of 27) were revealed to be nonmalignant lesions. In this latter group the response of 3 specimens was false negative on histologic inspection, 2 showing cancer cells without surface infiltration. We thus conclude that this dye-scattering method is of great use in enabling the differentiation of nonmalignant mucosa from neoplastic lesions and the preoperative determination of the stage of a tumor. PMID- 8420420 TI - Diagnosis of pneumonia in mechanically ventilated patients. Repeatability of the protected specimen brush. AB - The repeatability (i.e., the variation in repeated measurements of the same quantity) of the protected specimen brush (PSB) with quantitative cultures was assessed in 22 consecutive mechanically ventilated (MV) patients with suspected nosocomial pneumonia. Five PSB samples were collected in the same lung area during the same bronchoscopic procedure and processed for bacteriologic identification and quantitative culture. A laboratory control was also performed in order to assess the in vitro repeatability of the quantitative culture technique. The five PSB always recovered the same microorganisms, indicating a 100% qualitative repeatability for the PSB. Conversely, the quantitative repeatability was somewhat lower since in 59% of the patients the quantitative results varied by more than one log10, which is the minimal precision affordable with quantitative cultures. The distinction between presence or absence of infection based on the 10(3) cfu/ml recommended diagnostic threshold was, however, only moderately affected by the variability of the quantitative results since only three of 22 patients (13.6%) displayed results spread out on each side of the 10(3) cfu/ml break point. Intrasubject variability of quantitative results was not explained by problems with the quantitative culture technique, which proved excellent repeatability in the laboratory. This study indicated that, although the PSB technique with quantitative cultures displays an acceptable level of repeatability, caution is advisable when interpreting PSB results in critically ill patients with suspected pneumonia, especially if one refers to the 10(3) cfu/ml recommended diagnostic threshold and if a decision to treat or to abstain from treating is to be made. PMID- 8420421 TI - Inhibition of platelet-activating factor-induced bronchoconstriction by the leukotriene D4 receptor antagonist ICI 204,219. AB - We determined whether platelet-activating factor (PAF) caused bronchoconstriction through the release of leukotrienes (LT) by studying the effect of a potent LTD4 receptor antagonist, ICI 204,219, on PAF-induced bronchoconstriction in normal subjects. Eight males (age 20 to 36 yr) were given either 40 mg ICI 204,219 orally or matching placebo on 2 study days separated by 2 wk in a double-blind crossover study. Specific airway conductance (SGaw) was measured after inhalation of nebulized PAF (45 micrograms). ICI 204,219 caused a significant inhibition of PAF-induced bronchoconstriction as assessed by comparison of the maximum fail in SGaw or of the area under the curve of the time course of SGaw. ICI 204,219 did not inhibit PAF-induced neutropenia. PAF-induced bronchoconstriction is mediated largely by the release of sulfidopeptide leukotrienes in normal men. PMID- 8420422 TI - Surfactant and the adult respiratory distress syndrome. AB - ARDS includes a complex series of events leading to alveolar damage, high permeability pulmonary edema, and respiratory failure. The endogenous pulmonary surfactant system is crucial to maintaining normal lung function, and only recently has it been appreciated that alterations in the surfactant system significantly contributed to the pathophysiology of the lung injury of patients with ARDS. Through a combination of analyzing BAL samples from patients with ARDS and extensive animal studies, there have been significant insights into the variety of surfactant abnormalities that can occur in injured lungs. These include altered surfactant composition and pool sizes, abnormal surfactant metabolism, and inactivation of alveolar surfactant by serum proteins present within the airspace. Positive effects of exogenous surfactant administration on acute lung injury have been reported. There is now a prospective, randomized clinical trial evaluating the efficacy of aerosolized exogenous surfactant in patients with ARDS. This trial has demonstrated improvements in gas exchange and a trend toward decreased mortality in response to the surfactant. Despite these encouraging results, there are multiple factors requiring further investigation in the development of optimal surfactant treatment strategies for patients with ARDS. Such factors include the development of optimal surfactant delivery techniques, determining the ideal time for surfactant administration during the course of injury, and the development of optimal exogenous surfactant preparations that will be used to treat these patients. With further clinical trials and continued research efforts, exogenous surfactant administration should play a useful role in the future therapeutic approach to patients with ARDS. PMID- 8420423 TI - Neuromuscular blockade in the intensive care unit. More than we bargained for. PMID- 8420424 TI - Pulmonary function of U.S. coal miners related to dust exposure. PMID- 8420426 TI - State of the ARRD. PMID- 8420425 TI - Oxygen consumption is independent of increases in oxygen delivery by dobutamine in septic patients who have normal or increased plasma lactate. AB - We asked whether the relationship between oxygen delivery and oxygen consumption is different between patients who have sepsis and normal (n = 6) or increased (n = 8) concentrations of plasma lactate. We determined oxygen consumption using analysis of respiratory gases while increasing oxygen delivery using a dobutamine infusion. The relationship between oxygen delivery and consumption was y = 124 + 0.043 * x in the normal lactate group and y = 131 - 0.003 * x in the high lactate group (95% CI for differences in slopes, -0.003 to 0.096; p < or = 0.05 for slope, normal versus high lactate). In the normal lactate group, direct oxygen consumption increased by only 8 +/- 6 ml/min/m2 after dobutamine infusion (from 144 +/- 26 to 153 +/- 22 ml/min/m2, p < or = 0.02) despite an average increase of 220 +/- 80 ml/min/m2 in oxygen delivery (from 446 +/- 91 to 666 +/- 90 ml/min/m2, p < or = 0.01). The oxygen extraction ratio fell from 0.27 +/- 0.03 to 0.21 +/- 0.02 after dobutamine (p < or = 0.017). In the high lactate group, direct oxygen consumption decreased by 1 +/- 6 ml/min/m2 after dobutamine (from 131 +/- 33 to 130 +/- 35 ml/min/m2, p > 0.60) despite an average increase of 168 +/- 138 ml/min/m2 in oxygen delivery (from 467 +/- 194 to 635 +/- 300 ml/min/m2, p < or = 0.01). The oxygen extraction ratio fell from 0.30 +/- 0.14 to 0.26 +/- 0.12 after dobutamine (p < or = 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420427 TI - Effect of sleep on respiratory muscle activity during mechanical ventilation. AB - The purpose of this study was to determine whether consciousness was critical for the expression of neuromechanical inhibition of breathing during mechanical ventilation. This same mechanical ventilation model also was used to evaluate the relative importance of sleep state in causing CO2 retention during sleep. Positive pressure ventilation was used to suppress respiratory muscle activity; CO2 was then added until a reappearance of inspiratory effort, which defined the recruitment threshold (PCO2RT). Keeping the mechanics of the respiratory system constant through the use of passive mechanical ventilation allowed us to measure the output of the respiratory controller, independent of these parameters. Eight normal subjects were mechanically hyperventilated with a nasal mask during wakefulness and sleep with matched flow rates, frequencies, and tidal volumes. When inspiratory muscle activity was undetectable and end-tidal PCO2 (PETCO2) fell below 30 mm Hg, inspired CO2 was added in stepped increments until inspiration reoccurred. The sleeping state increased both eupneic PETCO2 (42 +/- 4 versus 38 +/- 3 mm Hg) and PCO2RT (48 +/- 3 versus 46 +/- 2 mm Hg) compared with that during wakefulness. Neuromechanical inhibition of inspiratory muscle activity during mechanical ventilation was present during both wakefulness and sleep, as evidenced by the mean difference between PCO2RT and eupneic PETCO2 of 8 and 6 mm Hg, respectively. Recruitment thresholds during wakefulness and sleep were compared to evaluate the effect of sleep on respiratory motor output independent of changes in load, i.e., respiratory mechanics held constant.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420428 TI - Follow-up protected specimen brushes to assess treatment in nosocomial pneumonia. AB - To prospectively determine the bacteriologic and clinical efficacy of antimicrobial therapy for nosocomial bacterial pneumonia selected based upon information provided by cultures of protected specimen brush (PSB) samples obtained during bronchoscopy, 76 consecutive patients with ventilator-associated pneumonia were studied using follow-up quantitative PSB cultures obtained after 3 days of treatment. Of the 173 microorganisms initially present in the PSB samples, only 11 (6%) were not eradicated by antimicrobial therapy, including three recovered at high (> or = 10(3) cfu/ml) concentrations. Thirty-two emerging pathogens, including nine at high concentrations, were also detected; 26 of them (81%) were resistant to the initial antibiotics administered. Of the 76 patients included in the study, cultures of follow-up PSB samples identified 51 in whom the infection site in the lung was completely sterilized, 16 with low-grade infection, and only nine with persistent high-grade infection. Analysis of clinical outcome within the 15 days after the initiation of antimicrobial therapy demonstrated clinical improvement in 62 of 67 (93%) patients in whom the site of infection was contained by treatment as compared with four of 9 (44%) patients with persistent high-grade infection (p < 0.01). These data suggest that appropriate antimicrobial therapy for ventilator-associated pneumonia results in the control of the initial infection in 88% of the patients. However, an early superinfection caused by multiresistant pathogens can occur in a small subset of these patients. When follow-up PSB cultures were negative, an improved outcome was noted. PMID- 8420429 TI - Importance of inspiratory load in the assessment of severity of airways obstruction and its correlation with CO2 retention in chronic obstructive pulmonary disease. AB - Different aspects of mechanical loading were analyzed in a group of 25 patients with chronic obstructive pulmonary disease (COPD) who all had severe expiratory airflow limitation but different arterial CO2 values. It was found that the maximal expiratory flow rates (V75, V50, V25, VFRC), functional residual capacity (FRC), residual volume (RV), total lung capacity (TLC), specific airways conductance (SGaw) measured during inspiration or expiration, and lung recoil pressure (PLel) were not correlated to the PaCO2. Only five parameters were significantly linked to the PaCO2; they were the peak inspiratory flow (PIF) (r = -0.57, p < 0.002), the inspiratory vital capacity (IVC) (r = -0.46, p < 0.01), the maximal voluntary ventilation (MVV) (r = -0.49, p < 0.01), the total SGaw (r = -0.40, p < 0.03), and the forced expiratory flow volume in one second (FEV1) (r = -0.36, p < 0.05). It was concluded that the airflow limitation during the inspiratory phase has an important contribution to the CO2 retention in patients with COPD, and therefore analysis of both phases of the respiratory cycle is necessary to assess the severity of airway obstruction in this disease. PMID- 8420430 TI - Physiologic effects of positive end-expiratory pressure in patients with chronic obstructive pulmonary disease during acute ventilatory failure and controlled mechanical ventilation. AB - Dynamic hyperinflation and intrinsic positive end-expiratory pressure (PEEPi) are observed in patients with chronic obstructive pulmonary disease (COPD) and flow limitation. Several reports suggest that PEEP levels approaching PEEPi reduce inspiratory load due to PEEPi, without further hyperinflation. Hence PEEP should not increase intrathoracic pressure or affect hemodynamics and gas exchange. To verify this hypothesis, the effects of PEEP (0 to 15 cm H2O) on respiratory mechanics, hemodynamics, and gas exchange were studied in nine COPD patients during controlled mechanical ventilation. PEEP levels approaching PEEPi (9.8 +/- 0.5 cm H2O) did not affect the expiratory flow/volume relationship, confirming the presence of flow limitation. PEEP levels of 5 and 10 cm H2O did not change lung volume and PEEPi in the respiratory system (PEEPtot,rs) and chest wall (PEEPtot,cw) or affect hemodynamics and gas exchange. When applied PEEP overcame PEEPi, changes in lung volume and the expiratory flow/volume relationship were observed. PEEPtot,rs and PEEPtot,cw also increased. Under these circumstances, PEEP increased static elastance in both the respiratory system and the chest wall, reducing cardiac index and affecting hemodynamics and gas exchange. Our data show that in mechanically ventilated COPD patients with PEEPi due to flow limitation, PEEP levels exceeding the 85% of PEEPi (Pcrit) caused further hyperinflation and compromised hemodynamics and gas exchange. PMID- 8420431 TI - A comparison of clinical assessment and home oximetry in the diagnosis of obstructive sleep apnea. AB - In order to determine whether measurement of arterial oxygen saturation (SaO2) could identify patients with obstructive sleep apnea (OSA), 98 consecutive patients referred for assessment of snoring and/or daytime somnolence were assessed clinically and then underwent both unsupervised oximetry in their homes and formal polysomnography. Clinical assessment identified patients with an apnea+hypopnea index (AHI) > or = 15 events per hour with a sensitivity of 79% and a specificity of 50%. Home oximetry analyzed by counting the number of arterial oxygen desaturations recorded was inferior to clinical assessment. For desaturations of 2% or more from baseline, desaturation index (DI) > or = 15 per hour identified patients with AHI > or = 15 with sensitivity 65% and specificity 74%; for 3% desaturations, sensitivity was 51% and specificity 90%; and for 4% desaturations, sensitivity was 40% and specificity 98%. From the oximetry data, the percentage of time spent at SaO2 below 90% (CT90) was also calculated. CT90 > or = 1% identified patients with AHI > or = 15 with sensitivity 93% and specificity 51%; for patients with AHI > or = 15 ultimately given nasal continuous positive airway pressure (CPAP), the sensitivity of a CT90 > or = 1% was 100%. We concluded that home oximetry with CT90 < 1% practically excludes clinically significant OSA. Conversely, home oximetry with DI > or = 15 for 4% desaturations makes OSA likely: the positive predictive value for OSA is 83% if the pretest probability of OSA is 30% and over 90% if the pretest probability is at least 50%. PMID- 8420432 TI - Steady-state ventilatory responses to expiratory resistive loading in quadriplegics. AB - Patients with quadriplegia have a limited capacity to recruit expiratory muscles and are deprived of respiratory-related feedback from the rib cage and abdominal wall. We wished to evaluate the compensatory strategies available to such patients during expiratory resistive loading (ERL) and to compare their responses with those of normal healthy individuals. In addition, to determine whether the quadriplegic subjects have a blunted sensory appreciation of added ERL, we also compared sensory detection thresholds (delta R50). Steady-state ventilatory responses to ERL (delta R = 12 cm H2O/L/s) were compared in seven quadriplegic patients (level of injury, C6, C7) and six age-matched normal subjects. Highly significant intergroup differences were evident in the extent of prolongation of expiratory time (TE) and total cycle duration (Ttot) during ERL; values of delta TE and delta Ttot in quadriplegics were, on average, 46% of those of normals (p < 0.001). Minute ventilation (VE) was defended to an equal or better extent in quadriplegics. ERL-induced changes in tidal volume, inspiratory duration, mean inspiratory and expiratory flows, and end-expiratory lung volume (EELV) were not significantly different. Average delta R50 in quadriplegics and normals were (mean +/- SD), 1.73 +/- 0.039 cm H2O/L/s and 1.62 +/- 0.4 cm H2O/L/s, respectively (p = ns). Quadriplegics, therefore, despite substantial sensory and motor deficits, defend ventilation and EELV as effectively as normal individuals and show no attenuation in the ability to detect an added expiratory resistance. PMID- 8420433 TI - Effects produced by infusion of a free radical-generating solution into the diaphragm. AB - Although studies have examined the susceptibility and pattern of injury induced by infusion of free radical-generating solutions into a number of vital organs, no such investigation has been performed for the diaphragm. The purpose of the present study was to examine the susceptibility of the diaphragm to damage by a free radical-generating solution (iron-ADP complexes). Studies were performed using an in situ canine diaphragmatic strip preparation in which the phrenic artery supplying the strip was cannulated and perfused with blood from the ipsilateral femoral artery. Four groups of studies were performed: (1) a group in which saline was infused into the arterial supply of the diaphragm for 15 min; (2) a group in which a solution of iron-ADP was infused; (3) a group in which both iron-ADP and superoxide dismutase (SOD), a free radical scavenger, were infused; and (4) a group given iron-ADP and denatured SOD. Strip tension and blood flow were monitored during electrically induced diaphragmatic contractions for 15 min before intraphrenic infusions, during the period of infusions, and for 90 min after cessation of infusions. We found that diaphragm tension did not change over time in saline-treated control animals but fell significantly in animals in which iron-ADP was infused. The effects of iron-ADP were largely prevented by concomitant administration of active SOD, but not by denatured SOD. On average, at 90 min after cessation of infusions, tension had fallen to 82 +/- 6, 41 +/- 8, 63 +/- 4, and 28 +/- 9% of its initial value in saline, iron-ADP, iron-ADP/SOD, and iron-ADP/denatured SOD groups, respectively (p < 0.001 for comparison of the four groups, with saline and iron-ADP/SOD groups different from the other two groups). Diaphragm blood flow did not change significantly in any group. These data suggest that free radical-mediated diaphragmatic injury can result in a marked reduction in diaphragm contractility. PMID- 8420434 TI - Diaphragmatic dysfunction in neuralgic amyotrophy: an electrophysiologic evaluation of 16 patients presenting with dyspnea. AB - We report 16 adult men (age, 41 to 75 yr) with neuralgic amyotrophy (NA) who presented with dyspnea due to involvement of the diaphragm. All patients developed breathlessness after a prodrome of acute severe neck and shoulder pain. Bilateral diaphragm paralysis (BDP) was confirmed in 12 patients and unilateral diaphragm paralysis (UDP) in four by the absence of electrical and mechanical responses to percutaneous phrenic nerve stimulation. Global expiratory muscle strength was well preserved in all patients, but inspiratory muscle strength was reduced in proportion to the extent of diaphragmatic involvement. Lung function showed low lung volumes with preservation of carbon monoxide transfer coefficient in all patients. Two BDP patients were hypoxic (PaO2 = 67 and 54 mm Hg, respectively) on daytime arterial blood gas analysis; the latter patient with pre existing chronic obstructive pulmonary disease and marked obesity also had borderline hypercapnia (PaO2 = 49 mm Hg). Overnight sleep studies in three BDP and two UDP patients showed frequent intermittent arterial oxygen desaturations apparently caused by obstructive sleep apneas, but there was no evidence of alveolar hypoventilation. Follow-up muscle studies in five BDP and four UDP patients between 2 and 4 yr after initial referral showed complete recovery of diaphragmatic function in only two UDP patients, one of whom relapsed a year later. We postulate that NA may be an important but underrecognized cause of diaphragmatic paralysis in otherwise normal patients. Diaphragmatic strength returns very slowly, if at all. PMID- 8420435 TI - Hypoplastic trachea in Down's syndrome. AB - Our study sought to determine whether tracheal dimensions were reduced in patients with Down's syndrome and whether such a narrowing, if present, was related to the presence of congenital heart disease or to body habitus (height or weight). The inner diameter of the tracheal air column was measured at 2 cm above the aortic arch in 14 adult patients with Down's syndrome, and the results compared with previously established norms. Two-way analysis of variance showed no significant difference in z-scores between sexes or between those with or without congenital heart disease. Z-scores were significantly different from zero for both coronal diameters (p = 0.0010) and sagittal diameters (p = 0.0003). The negative limits on the 95% confidence interval for coronal z-scores (-2.5 to 0.8) and sagittal z-scores (-2.6 to -1.0) indicate that our patients have tracheal diameters significantly smaller than normal. Linear regression analysis showed no significant correlation between tracheal diameters and patients' height or weight. We conclude that tracheal diameters in adult patients with Down's syndrome are reduced and that the narrowing cannot be ascribed to associated congenital heart disease or to body habitus. PMID- 8420436 TI - Bronchial lavage and bronchoalveolar lavage in allergen-induced single early and dual asthmatic responders. AB - The phenotypic cellular profile of bronchial lavage (BL) and bronchoalveolar lavage (BAL) was studied in 7 single early (SR) and 10 dual asthmatic responders (DR). Lavage was performed, after previously having determined bronchial hyperresponsiveness to histamine and the response to house dust mite (HDM) challenge. The recovered lavage fluid was separated in two fractions, BL and BAL. Total fluid recovery and cell number from the BL and BAL were comparable in both patient groups. Differential cell counting and immunocytochemistry were performed. DR had a significantly higher number of eosinophils and EG2+ cells in BL but not in their BAL. No differences could be found in CD4+, CD8+, and HLA-DR+ cells. A strong correlation was found between eosinophils in the BL+ and EG2+ cells in the BL (r = 0.79, p < 0.001) and between eosinophils in the BL and peripheral blood eosinophils (r = 0.70, p < 0.0025). The number of EG2+ cells and the number of epithelial cells in both BL and BAL showed a correlation (r = 0.55, p < 0.05). Dual responders had a higher total IgE (p < 0.01), and total serum IgE correlated well with the eosinophils in the BL (r = 0.85, p < 0.0001). Our observations demonstrate cellular differences in the lung on mainly a bronchial level between single early and dual asthmatic responders. A bronchial lavage eosinophil and EG2+ cell count and higher blood total IgE level are associated with the tendency to develop a dual asthmatic response. PMID- 8420437 TI - Effect of the platelet-activating factor antagonist UK-74,505 on the early and late response to allergen. AB - The effect of UK-74,505, a specific platelet-activating factor (PAF) antagonist, on the early (EAR) and late asthmatic response (LAR) to inhaled allergen was studied in a randomized, double-blind, placebo-controlled crossover study. A total of eight adult male atopic asthmatic subjects completed the protocol (one withdrew after screening), all having demonstrated a dual response to inhaled allergen (EAR, > 20% fall in FEV1 between 0 and 1 h; LAR, > 15% fall in FEV1 between 4 and 8 h after challenge). Subjects were studied on 2 days at least 10 days apart. After measurement of baseline FEV1, subjects ingested a single oral dose of 100 mg UK-74,505 or matched placebo (P). Allergen challenge was performed 3 h later and the FEV1 was then measured for 8 h. There was no difference between UK-74,505 and placebo in the maximum percentage change from baseline (+/- SEM) for either EAR or LAR (EAR, UK-74,505 -25.6 +/- 4.8%, P -24.0 +/- 3.3%; LAR, UK 74,505 -20.8 +/- 4.4%, P -25.7 +/- 3.8%). There was no significant difference in the area under the percentage change from baseline FEV1-time curve. Ex vivo platelet aggregation to PAF was measured at 0, 2, 6, 8, and 10 h after the dose. There was marked inhibition of platelet aggregation to PAF for 10 h following UK 74,505 but not placebo (% maximum aggregation to PAF, UK-74,505, -69.9%; P, 0.13%; p = 0.0001). Histamine challenge was performed in five patients the day before and after each study day. There was no significant difference between UK 74,505 and placebo in PD20 to histamine (mean PD20 before and after UK-74,505, 1.31 and 0.96 mumol; P, 1.32 and 1.17 mumol). UK-74,505 did not affect either the EAR or the LAR to inhaled allergen or bronchial responsiveness, despite its potency and long duration of action. This suggests that PAF does not have a major role in the acute response to inhaled allergen. PMID- 8420438 TI - Increased granulocyte/macrophage colony-stimulating factor production by mononuclear cells from peripheral blood of patients with bronchial asthma. AB - The in vitro production of granulocyte/macrophage colony-stimulating factor (GM CSF) by mononuclear cells (MNC) from the peripheral blood of patients with bronchial asthma (BA) was examined by enzyme-linked immunosorbent assay (ELISA). GM-CSF concentrations in the media of MNC from patients with BA cultured with interleukin-2 (IL-2) was 80.2 +/- 52.0 pg/ml (mean +/- SE, n = 12), and that in cultures without a stimulant was 12.1 +/- 11.3 pg/ml. The GM-CSF concentrations in the media of MNC from patients with other diseases (n = 13) and from healthy volunteers (n = 6) cultured with or without IL-2 were less than 7.5 pg/ml (the minimum detectable value). The culture media of MNC from patients with BA demonstrated activities for stimulating the proliferation and survival of eosinophils, and these activities were partially inhibited by anti-GM-CSF antibodies. GM-CSF production by MNC of patients with BA treated with glucocorticoids was lower than that of MNC from untreated patients with BA, and it was inhibited by coculture with glucocorticoids in vitro. These results suggest that GM-CSF production by MNC is increased in patients with BA, is modulated by glucocorticoids, and may play an important role in the pathogenesis of BA. PMID- 8420439 TI - Histamine N-methyltransferase modulates histamine- and antigen-induced bronchoconstriction in guinea pigs in vivo. AB - To examine whether histamine N-methyltransferase (HMT; EC 2.1.1.8) modulates the effects of allergic reaction in vivo, we studied the effects of aerosolized SKF 91488, a specific HMT inhibitor, on the responses to aerosolized histamine in unsensitized guinea pigs and to ovalbumin (OA) antigen inhalation in guinea pigs sensitized to OA. Airway responsiveness was assessed by determining provocation concentrations of histamine and OA aerosols that increased pulmonary resistance to twice the baseline values. SKF 94188 shifted, in a dose-dependent fashion, the dose-response curves to histamine and OA antigen to lower concentrations, and it significantly decreased provocation concentrations of both histamine and OA antigen (p < 0.01). In contrast of SKF 91488, aerosolized aminoguanidine, a specific inhibitor of diamine oxidase (10(-2) M, 90 breaths), did not alter the provocation concentration of histamine (p > 0.20). SKF 91488 (10(-2) M, 90 breaths) caused no significant changes in response to acetylcholine (p > 0.30). HMT activities were observed in the entire airways of the trachea, main bronchi, segmental bronchi and bronchioles, and parenchymal tissues. These findings suggest that HMT modulates the effects of exogenous histamine and endogenously released histamine by antigen challenge on bronchoconstrictor responses in guinea pigs in vivo. PMID- 8420440 TI - Effects of smoked substance abuse on nonspecific airway hyperresponsiveness. AB - Previous data suggest that regular tobacco smoking may lead to nonspecific airway hyperresponsiveness (AHR) independent of airway obstruction, possibly because of effects on bronchial inflammation or mucosal permeability. Little is known concerning the effects on AHR of other widely smoked substances besides tobacco such as marijuana or cocaine. The smoke of both marijuana and cocaine contains respiratory irritants that elicit cough and produce abnormalities in airway dynamics and bronchial mucosal histopathology in habitual smokers. Therefore, regular smoking of one or both of these illicit substances could cause AHR or augment the AHR associated with tobacco smoking. The present study examined the influence of habitual smoking of marijuana, cocaine, and/or tobacco on nonspecific AHR in 542 (456 male) healthy participants (mean age, 34.8 +/- 6.8 SD yr) in an ongoing cohort study of the pulmonary effects of habitual smoking of illicit substances. Subjects with a history of intravenous drug abuse, significant occupational exposures, asthma, or recent upper respiratory tract infection were excluded. Inhalation challenge studies were performed using solutions of diluent and methacholine chloride (1.25 to 25 mg/ml) aerosolized by a DeVilbiss no. 646 nebulizer attached to a breath-activated dosimeter inhaled by three to five inspiratory capacity breaths. Positive responses to methacholine were defined by > or = 20% or > or = 10% declines in FEV1 from the postdiluent control value after inhalation of each concentration of methacholine. Participants were categorized by smoking status (nonsmoking and smoking of marijuana, cocaine, and/or tobacco alone and in combination); most analyses were performed in men and women separately.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420441 TI - Assessment of a retrovirus sequence and other possible risk factors for the chronic fatigue syndrome in adults. AB - OBJECTIVE: To assess whether the human T-lymphotropic virus type II (HTLV-II) gag gene sequence, a purportedly new laboratory marker of the chronic fatigue syndrome (CFS), and other possible risk factors for CFS, particularly those associated with retroviral transmission, are associated with well-characterized CFS. DESIGN: Two matched case-control studies. SETTING: The metropolitan Atlanta area. PATIENTS: Twenty-one patients with CFS who were identified by the Centers for Disease Control and Prevention CFS surveillance system; 21 CDC employee controls (laboratory study) and 42 neighborhood controls (risk-factor study) who were matched to patients by age, race, and gender. MEASUREMENTS: Peripheral blood lymphocytes and leukocytes were assayed for the HTLV-II gag gene sequence by polymerase chain reaction and specific Southern blot hybridization. Questionnaires elicited demographic and clinical information and a history of exposures associated with retrovirus transmission (for example, blood transfusions, sexual practices, intravenous drug use). RESULTS: All patients were white and 86% were female. The median age at illness onset was 34 years (range, 16 to 51 years). The HTLV-II gag gene sequence was not identified in the blood of any patient or control under conditions in which the appropriate assay controls were positive. No statistical differences were observed between patients and controls in frequency of blood transfusions (10% compared with 7%), median number of sex partners before illness (3 compared with 3), bisexual or homosexual behavior (14% compared with 7%), intravenous drug use (0% compared with 0%), and other factors associated with retroviral infection. CONCLUSIONS: The HTLV-II gag gene sequence was not a marker for CFS in this small study of well-defined patients, nor did other characteristics of the patients and controls support the hypothesis that a retrovirus, transmitted by usual modes, was a cause of CFS. PMID- 8420442 TI - Improvement of lipid abnormalities associated with proteinuria using fosinopril, an angiotensin-converting enzyme inhibitor. AB - OBJECTIVE: To examine whether reducing protein excretion in patients with proteinuric renal disease using an angiotensin-converting enzyme inhibitor, fosinopril sodium, would be accompanied by an amelioration of the associated hyperlipidemia. DESIGN: A randomized, placebo-controlled, double-blind study of 12 weeks, followed by 23 weeks of an open-label trial using fosinopril. SETTING: Outpatient renal clinics. PATIENTS: Twenty-six patients (age range, 28 to 70 years) with mild to moderate renal impairment and proteinuria associated with type II diabetes (15 patients) and other causes of nondiabetic renal disease (11 patients) completed the double-blind phase of the study. All patients except one were men. INTERVENTION: Fosinopril, 10 mg initial oral daily dose (randomized trial), and 20 mg orally once a day (open-label phase). MEASUREMENTS: Proteinuria and serum lipids (total cholesterol, high-density lipoprotein, and low-density lipoprotein [LDL] cholesterol, and lipoprotein(a) protein). RESULTS: In a group of 17 patients treated with fosinopril, protein excretion decreased from 5.56 to 4.28 g/d, a reduction of 1.28 (95% CI, -2.49 to -0.08). The reduction was associated with a decrease in serum total cholesterol from 6.39 to 5.82 mmol/L, a decrease of 0.58 mmol/L (CI, -1.01 to -0.15 mmol/L). In a group of nine patients treated with placebo, neither protein excretion (from 5.11 to 4.81 g/d, a change of -0.29 g/d [CI, -1.78 to +1.13 g/d]) nor serum total cholesterol (from 6.08 to 5.77 mmol/L, a change of -0.31 mmol/L [CI, -0.78 to +0.13 mmol/L]) change significantly. At the end of the double-blind phase, plasma lipoprotein(a) protein decreased in the fosinopril-treated group (from 3.94 to 3.33 mg/dL, a reduction of 0.60 mg/dL [CI, -1.02 to -0.18 mg/dL]) but not in the placebo group (from 2.85 to 3.19 mg/dL, a change of +0.34 mg/dL [CI, -0.53 to +1.2 mg/dL]). Dietary protein and fat intake were similar in the two groups throughout the study. In 16 patients who completed an extended open-label phase, fosinopril was associted with a decrease in protein excretion from 4.53 to 3.22 g/d, a reduction of 1.29 g/d (CI, -2.54 to -0.05 g/d), which was associated with a reduction in serum total cholesterol (from 6.37 to 5.54 mmol/L, a decrease of 0.84 mmol/L [CI, -1.59 to -0.08 mmol/L]), LDL cholesterol (from 4.38 to 3.72 mmol/L [a decrease of 0.68 mmol/L [CI, -1.33 to -0.03 mmol/L]), and plasma lipoprotein(a) protein (from 3.58 to 2.81 mg/dL, a reduction of 0.82 mg/dL [CI, -1.58 to -0.05 mg/dL]). CONCLUSION: The angiotensin-converting enzyme inhibitor, fosinopril, can result in a sustained reduction in serum total cholesterol, LDL cholesterol, and plasma lipoprotein(a) protein levels in conjunction with a partial reduction in proteinuria. PMID- 8420444 TI - Antihypertensive drug therapy and the initiation of treatment for diabetes mellitus. AB - OBJECTIVE: To quantify the risk for the occurrence of hyperglycemia requiring initiation of therapy among patients taking various antihypertensive regimens. DESIGN: Case-control study. SETTING: New Jersey Medicaid program. PATIENTS: The study included New Jersey Medicaid enrollees 35 years of age or older. The 11,855 case patients were newly started on a hypoglycemic agent (oral agent or insulin) between 1981 and 1990. The 11,855 controls were selected randomly from among other Medicaid enrollees. MEASUREMENTS AND MAIN RESULTS: The frequency of initiation of hypoglycemic therapy was increased for users of virtually all antihypertensive agents relative to nonusers after adjustment for age, gender, race, nursing home residency, number of days hospitalized, total number of prescriptions, and selected medication exposures. The estimated relative risk for initiation of hypoglycemic therapy was 1.40 for patients receiving thiazide diuretics (95% CI, 1.26 to 1.58) and ranged from 1.56 to 1.77 for patients receiving other antihypertensive medications, depending on the medication category. A higher risk was associated with multiple-agent regimens, whether they excluded a thiazide diuretic (odds ratio, 1.76; CI, 1.49 to 2.07) or included one (odds ratio, 1.93; CI, 1.75 to 2.13). When the analysis was restricted to users of antihypertensive agents (n = 8005), the risk associated with other single agent antihypertensive regimens was not significantly different from that associated with thiazide diuretics. However, patients receiving multiple-agent regimens continued to be at increased risk for hyperglycemia requiring hypoglycemic therapy relative to those who used thiazide diuretic therapy alone. CONCLUSION: The association between antihypertensive therapy and the initiation of treatment for diabetes mellitus is more closely related to the intensity of therapy than to the individual agent used. Our data do not support the hypothesis that thiazide diuretics are more strongly associated with the initiation of hypoglycemic therapy than are other antihypertensive agents. PMID- 8420443 TI - Veno-occlusive disease of the liver and multiorgan failure after bone marrow transplantation: a cohort study of 355 patients. AB - OBJECTIVE: To determine the incidence and clinical course of veno-occlusive disease of the liver (VOD) after bone marrow transplantation and to analyze risk factors for severe VOD. DESIGN: Cohort study of 355 consecutive patients. SETTING: A bone marrow transplantation center. MEASUREMENTS: Each patient was prospectively evaluated for VOD, and many risk factors for severe VOD were analyzed using logistic regression models. The relation of VOD to renal and cardiopulmonary failure was analyzed using time-dependent proportional hazards models. RESULTS: Veno-occlusive disease developed in 190 of 355 patients (54%; 95% CI, 48% to 59%): Fifty-four patients had severe VOD and 136 had mild or moderate VOD. Independent variables derived from a multivariate model for predicting severe VOD included elevated transaminase values before transplantation (relative risk, 4.6; P < 0.0001); vancomycin therapy during cytoreductive therapy (relative risk, 2.9; P = 0.003); cytoreductive therapy with a high-dose regimen (relative risk, 2.8; P = 0.01); acyclovir therapy before transplantation (relative risk, 4.8; P = 0.02); mismatched or unrelated donor marrow (relative risk, 2.4; P = 0.02); and previous radiation therapy to the abdomen (relative risk, 2.2; P = 0.04). Vancomycin therapy was a marker for persistent fever. Multiorgan failure was more frequent among patients with VOD and usually followed the onset of liver disease. CONCLUSIONS: Veno-occlusive disease, which developed in 54% of bone marrow transplant recipients, is frequently associated with renal and cardiopulmonary failure. Pretransplant transaminase elevations, use of high-dose cytoreductive therapy, and persistent fever during cytoreductive therapy are independent predictors of severe VOD. PMID- 8420445 TI - Aminophylline for bradyasystolic cardiac arrest refractory to atropine and epinephrine. PMID- 8420447 TI - Medical consultation on surgical services: an annotated bibliography. PMID- 8420446 TI - Fine-needle aspiration biopsy of the thyroid: an appraisal. AB - OBJECTIVE: To review the literature on the utility of fine-needle aspiration biopsy in the diagnostic management of nodular thyroid disease. DATA SOURCES: Relevant articles published in major English-language medical journals during the last 10 years. DATA EXTRACTION: Articles were reviewed to assess the results of fine-needle aspiration biopsy and its effect on thyroid management and cost of care. DATA SYNTHESIS: Fine-needle aspiration biopsy of the thyroid gland is safe, inexpensive, minimally invasive, and highly accurate in the diagnosis of nodular thyroid disease. Four cytologic diagnostic categories are used. Rates for these categories, based on data pooled from seven series, were as follows: benign, 69%; suspicious, 10%; malignant, 4%; and nondiagnostic, 17%. Analysis of recent data suggests a false-negative rate of 1% to 11%, a false-positive rate of 1% to 8%, a sensitivity of 65% to 98%, and a specificity of 72% to 100%. Limitations of fine needle aspiration are related to the skill of the aspirator, the expertise of the cytologist, and the difficulty in distinguishing some benign cellular adenomas from their malignant counterparts. The introduction of fine-needle aspiration has had a substantial effect on the management of patients with thyroid nodules. The percentage of patients undergoing thyroidectomy has decreased by 25%, and the yield of carcinoma in patients who undergo surgery has increased from 15% to at least 30%. Fine-needle aspiration has decreased the cost of care by 25%. CONCLUSIONS: Fine-needle aspiration biopsy is safe, accurate, and cost-effective. The procedure has a central role in the management of thyroid nodules and should be used as the initial diagnostic test. PMID- 8420448 TI - A reappraisal of hepatitis B virus vaccination strategies using cost effectiveness analysis. AB - OBJECTIVE: To determine clinical and economic consequences of alternative vaccination strategies for preventing hepatitis B virus infection (HBV). METHODS: Decision analysis was used to evaluate costs, outcomes, and cost-effectiveness of three HBV management strategies ("no vaccination," "universal vaccination," and "screen and vaccinate") in four populations (newborns, 10-year-old adolescents, a high-risk adult population, and the general adult U.S. population). Information on HBV incidence and prevalence, clinical course, and management of acute illness and chronic sequelae was obtained from the literature and a panel of experts. Actual payments (costs) were obtained from Blue Cross/Blue Shield and local pharmacies. Incremental cost-effectiveness was calculated from the perspective of the payer of medical care and subjected to sensitivity analysis. RESULTS: Vaccination (with or without screening) prevents more disease at somewhat increased cost than no vaccination for the neonatal, adolescent, and adult populations. Vaccination (with or without screening) is a dominant strategy in adult high-risk populations (lower cost and greater benefit than no vaccination). Optimal cost-effectiveness, with nonmonetary benefits not discounted, results if all pregnant women are screened for active HBV infection, and HBV vaccine and hepatitis B immune globulin are administered to babies born to mothers with positive screening tests. Then HBV vaccine is administered to all children at age 10 and again 10 years later (incremental cost-per-year-of-life-saved relative to the "no vaccination" strategy is $375). A strategy of universal newborn vaccination alone leads to an incremental cost-per-year-of-life saved of $3332. If adolescents are vaccinated at age 10, incremental cost-per-year-of-life saved is $13,938; for the general adult population, the incremental cost-per-year-of life saved of universal vaccination is $54,524. Discounting benefits will increase cost-per-year-of-life saved 7 to 12 times for all strategies. CONCLUSIONS: HBV vaccine is most cost-effective when a strategy of screening newborns is combined with routine administration to 10-year-old children. The means to achieve substantial improvements in the health of the public in a cost effective fashion are now available and should be pursued aggressively. PMID- 8420449 TI - A modern Doctor Schweitzer. PMID- 8420450 TI - Hepatitis B immunization strategies: expanding the target. PMID- 8420451 TI - Clinical relevance of gastrointestinal hormones: emerging interest in hypergastrinemia. PMID- 8420452 TI - Microalbuminuria, lipoproteins, and diabetic control. PMID- 8420453 TI - Hodgkin disease and HIV. PMID- 8420454 TI - Adverse effects of ACE inhibitors. PMID- 8420455 TI - Adverse effects of ACE inhibitors. PMID- 8420456 TI - Adverse effects of ACE inhibitors. PMID- 8420457 TI - Adenosine-related ventricular asystole. PMID- 8420458 TI - Amiodarone and ventricular arrhythmia suppression. PMID- 8420459 TI - Alpha-interferon and reversible hypertriglyceridemia. PMID- 8420460 TI - Wine consumption in the elderly. PMID- 8420461 TI - Internal medicine: stop reporting problems and start solving them. PMID- 8420462 TI - Effects of otitis media on extended high-frequency hearing in children. AB - Extended high-frequency (EHF) hearing was studied in children with and without histories of chronic or recurrent otitis media (OM). The EHF thresholds were found to have good test-retest repeatability. Children with OM histories had poorer EHF hearing than children without OM histories. The EHF hearing in OM children appeared to be related to OM severity. Children with residual tympanometric abnormalities had poorer EHF hearing than OM children with normal middle ear function. The results suggest evidence for middle ear and inner ear components of EHF hearing losses in children with OM. PMID- 8420463 TI - Kikuchi-Fujimoto disease of the neck. Update. AB - Kikuchi-Fujimoto disease (KFD) is a rare entity of uncertain cause that commonly presents with persistently enlarged cervical lymph nodes unresponsive to antibiotic therapy. Although it usually follows a benign course, KFD has been repeatedly misdiagnosed as malignant lymphoma; hence, clinicians and pathologists alike need to be aware of this disease entity. The newest developments regarding the etiology and course of KFD are presented through a review of the literature and a recent illustrative case. PMID- 8420465 TI - Laryngeal brain stem evoked response in the porcine model. AB - Exaggeration of normally protective laryngeal reflexes is thought to play a role in several disorders, including the sudden infant death syndrome. An analysis of brain stem neural activity following laryngeal stimulation may provide insight into the pathophysiology of pathologic laryngeal reflexes and help to identify individuals at risk for these disorders. The purpose of this study was to define the far-field brain stem activity following laryngeal stimulation in the porcine model. This activity has been termed the laryngeal brain stem evoked response and may represent a potentially useful and objective measure of the neuronal activity in the laryngeal reflex pathway. Electrical stimulation of the superior laryngeal nerve was performed in 14 mixed-breed piglets under a variety of physiologic conditions. A total of six positive and six negative discrete waves were detected, with mean latencies ranging from 1.24 to 7.16 milliseconds. Stimulations performed during hypoxic, hypercapneic, or hypocapneic conditions resulted in no significant differences in waveform latencies. There appears to be a reproducible, but somewhat variable, brain stem response elicited by superior laryngeal nerve stimulation that can be recorded via a far-field technique in the porcine model. PMID- 8420464 TI - Extraction of inner ear antigens for studies in inner ear autoimmunity. AB - The search for a diagnostic assay in patients with autoimmune inner ear disease has led to the preparation of antigens from the inner ear, a presumed target in this disorder. In order to standardize the antigen preparations currently being used in the Western blot immunoassay, we have examined several distinct extraction procedures that employ well-known solutions and detergents. Results of this investigation clearly show that antigens of interest (68 kd, 33 to 35 kd, and 32 kd) are optimally extracted with a detergent (0.5% sodium dodecyl sulfate) and that bovine serum albumin, a potential contaminant, can be removed in the water-soluble fractions. Purification of inner ear antigens by these methods will lead to more reproducible results in immunoblotting, as well as a greater opportunity to identify the mechanisms involved in autoimmune inner ear disease. PMID- 8420466 TI - Evaluation of brain collateral circulation by the transcranial color Doppler guided Matas' test. AB - For the determination of the indication for reconstruction of the carotid artery in patients with head and neck cancer, a color Doppler-guided Matas' test was performed to examine brain collateral circulation and blood flow of the middle cerebral artery (MCA) and internal carotid artery (ICA). Both SSA-270A convex (3.75 MHz) and linear (3.75 MHz and 2.5 MHz) array probes (Toshiba Co, Ltd) were used for this investigation. A temporary occlusion test (60 seconds) by digital pressure on the common carotid artery was performed on 30 patients with head and neck cancer. The results were classified into four groups according to the relative flow volume (percent blood flow volume before the test) of the MCA: group A, with a flow volume of 87% or more; group B, with a flow volume between 72% and 86%; group C, with a flow volume between 57% and 71%; and group D, with a flow volume of 56% or less. The relative flow volume of the MCA was correlated with the ICA stump pressure and single photon emission computed tomography during a balloon Matas' test (ICA occlusion). This test seems to be useful in predicting the risk of carotid resection without reconstruction and in determining the indication for reconstruction of the carotid artery. PMID- 8420467 TI - Cell kinetics study of upper aerodigestive tract squamous cell carcinoma using bromodeoxyuridine. AB - Thirty patients treated for squamous cell carcinoma of the upper aerodigestive tract were reviewed for a cell kinetics study using bromodeoxyuridine (BrdU). Fresh specimens were obtained from the primary site of the tumors and incubated in Dulbecco's modified Eagle's medium containing BrdU to label S-phase tumor cells. The tumor specimens were stained immunohistochemically with monoclonal antibody against BrdU. The results demonstrated that the BrdU labeling index (BrdU LI) in these cases (470.3 +/- 118.2) was greater than the BrdU LI in normal mucosa (162 +/- 52.4). In a comparison of the clinical data and histopathologic features, the BrdU LI showed an inverse correlation with patient age (r = -.558; p < .01) and a direct correlation with the frequency of mitoses (r = .411; p < .05). Although the significance of the BrdU LI as a prognostic factor cannot be determined at present, this technique provides a simple in vitro test that avoids the potential hazards of in vivo labeling or the use of radioactive materials. PMID- 8420468 TI - Mucosal folds in human eustachian tube: a hypothesis regarding functional localization in the tube. AB - Differences in the extent of mucosal folding in the superior versus the inferior half of the human eustachian tube (ET) lumen were studied by measuring, as it appeared in histologic cross section, the length of the mucosal margin in the pharyngeal portion, the midcartilaginous portion, the isthmus portion, and the osseous portion of each half of the ET in 16 normal ET temporal bone specimens from 16 individuals 12 to 70 years of age. The mucosal margin of the ET was significantly longer in the inferior half than in the superior half in all four portions (paired t test: pharyngeal portion t[15] = 3.59, p < .01; midcartilaginous portion t[15] = 3.52, p < .01; isthmus portion t[15] = 4.15, p < .01; osseous portion t[15] = 2.24, p < .05). These results indicate that the inferior half of the ET mucosa has more folds than the superior half, and that the surface area of the inferior half of the ET is larger than that of the superior half. This, together with the finding previously reported that secretory components of the ET such as glands and goblet cells are more richly distributed in the inferior (floor) portion than in the superior (roof) portion of the ET, may be related to the fact that the floor of the ET has an active role in clearance function. A hypothesis regarding localization of the ventilatory and clearance functions within the ET is proposed. PMID- 8420469 TI - Comparison of quantitative DNA measurements and cytomorphology in squamous cell carcinomas of the upper aerodigestive tract with and without lymph node metastases. AB - Cytologic smears of squamous cell carcinomas (UICC T2 through T4) from the mucosa of the upper aerodigestive tract without cervical lymph node metastases (n = 10) and with metastases (n = 20) were examined. Subsequently, the cytologic smears of the metastases (n = 20) obtained by fine-needle aspiration or following neck dissection were evaluated morphologically. Each specimen was then stained with Papanicolaou and Feulgen techniques, and quantitative DNA measurements were performed with an image analysis system. Morphologically, the primary tumors with metastases revealed an increased mean nuclear perimeter and mean nuclear area compared with tumors unassociated with metastases. On quantitative DNA measurements, the tumors with metastases had an increased DNA content, a higher 2c deviation index, and a higher DNA malignancy grade and increased number of aneuploid cells. The specimens obtained from the metastatic nodes yielded the highest values for mean DNA content, 2c deviation index, aneuploidy, and malignancy grade. The stem lines of metastasis were always nondiploid. These data indicate a positive correlation between aneuploid tumor cell clones in primary cancer and the manifestation of lymph node metastases. PMID- 8420470 TI - Measurement of mucosal wave propagation and vertical phase difference in vocal fold vibration. AB - Examination of the surface wave properties of the vocal fold mucosa is becoming an important part of assessment of vocal function. A key wave property is propagation velocity, which determines the phase delay between the upper and lower margins of the vocal folds. Excised canine larynges were used to measure this phase delay, and therewith propagation velocity. The motion of two flesh points was tracked stroboscopically. Differential displacements between the flesh points were matched to displacements of a model. A least-squared fit of the data to the model provided the numeric values of propagation velocity, which varied from 0.5 m/s to about 2.0 m/s, depending on fundamental frequency. The corresponding phase delay along the medial surface of the vocal folds varied from about 60 degrees/mm to 30 degrees/mm. PMID- 8420471 TI - Olfactory evoked potentials and contingent negative variation simultaneously recorded for diagnosis of smell disorders. AB - Objective diagnosis of olfaction can be performed by registration of cortical olfactory evoked potentials (OEP) and of contingent negative variation (CNV). The CNV is a negative voltage developing at the vertex after discrimination of one of two smells while the patient is expecting a second stimulus. By an adequate procedure, including a long time window for averaging (2.56 seconds) with appropriate filters, the two tests can be performed simultaneously in a single session of less than 10 minutes. Anosmia is determinable by both OEP and CNV, although CNV shows less variability. On the other hand, CNV requires attention and some cooperation of the patient. Parosmia is accessible by CNV only; two odor qualities presented in random order have to be distinguished. Hyposmia can also be detected; just above the discrimination threshold, CNV amplitudes tend to be large--even enhanced--whereas OEP amplitudes may still be undetectable. PMID- 8420472 TI - Accumulation of potassium in scala vestibuli perilymph of the mammalian cochlea. AB - Movements of potassium (K+) were monitored during perfusion of either the scala tympani (ST) or the scala vestibuli (SV) of the guinea pig cochlea with a solution containing 15 mmol/LK+. A highly asymmetric clearance of K+ was observed, with K+ rapidly being taken up from the ST and allowed to accumulate in the SV. Under some conditions the SV K+ concentration could exceed that in the perfused ST. These observations are believed to result from the distortion of passive K+ diffusion by the high circulating current of K+ that is part of the transduction process. Calculations are presented to demonstrate that circulating fluxes are of sufficient magnitude to generate the results observed. The high rate of circulating K+ current is probably also responsible for the difference in physiologic K+ concentration between the ST and SV, in which the ST perilymph K+ concentration is typically found to be half that of the SV. A clearance of K+ from the ST and its eventual accumulation in the SV could play a role in how the ear responds to abnormal ion concentrations, such as may occur in Meniere's disease. It is proposed that an accumulation of K+ in the SV would result in vestibular dysfunction that might contribute to the vestibular symptoms of the disease. PMID- 8420473 TI - Internal carotid artery simulating a middle ear mass. PMID- 8420474 TI - Solitary fibrous tumor. AB - First described as a pleural neoplasm, the solitary fibrous tumor has been reported in a number of extrapleural sites, including the upper respiratory tract. The neoplasm is of mesenchymal origin, exists in benign and malignant forms, and is a histopathologic diagnosis made after exclusion of other soft tissue neoplasms. None of the eight reported sinonasal and nasopharyngeal solitary fibrous tumors have been histologically or biologically malignant. Surgical excision appears to control the tumor at these sites. PMID- 8420475 TI - Interpretation of electrocochleography in Meniere's disease and normal subjects. PMID- 8420476 TI - TNM staging of laryngeal cancer. PMID- 8420477 TI - Cochlear pathology in presbycusis. AB - A survey of the temporal bone collection at the Massachusetts Eye and Ear Infirmary reveals 21 cases that meet the criterion for the clinical diagnosis of presbycusis. It is evident that the previously advanced concept of four predominant pathologic types of presbycusis is valid, these being sensory, neural, strial, and cochlear conductive. An abrupt high-tone loss signals sensory presbycusis, a flat threshold pattern is indicative of strial presbycusis, and loss of word discrimination is characteristic of neural presbycusis. When the increments of threshold loss present a gradually decreasing linear distribution pattern on the audiometric scale and have no pathologic correlate, it is speculated that the hearing loss is caused by alterations in the physical characteristics of the cochlear duct, and the loss is identified as cochlear conductive presbycusis. It is clear that many individual cases do not separate into a specific type but have mixtures of these pathologic types and are termed mixed presbycusis. About 25% of all cases of presbycusis show none of the above characteristics and are classified as indeterminate presbycusis. PMID- 8420478 TI - Painful distal erythema and thrombocytosis. Erythromelalgia secondary to thrombocytosis. PMID- 8420479 TI - Reappearance of quiescent scars. Sarcoidosis. PMID- 8420480 TI - Anetoderma. Anetoderma, the primary type. PMID- 8420481 TI - Hyperhidrotic, painful lesion. Eccrine angiomatous hamartoma. PMID- 8420482 TI - Aspiration therapy in steatocystoma multiplex. PMID- 8420483 TI - The diagnostic challenge of diabetic hands. PMID- 8420484 TI - Caution in the use of local therapies for Kaposi's sarcoma. PMID- 8420485 TI - The art and science of dermatology. PMID- 8420486 TI - Isotretinoin does prevent skin cancer. PMID- 8420488 TI - 'Recent' advances in dermatology, medicine, cardiology, surgery, and rheumatology. PMID- 8420487 TI - Bullous variant of chemotherapy-induced acral erythema. PMID- 8420489 TI - Management of lentigo maligna. PMID- 8420490 TI - Cardiofaciocutaneous syndrome. PMID- 8420491 TI - Controlled trial of azathioprine and plasma exchange in addition to prednisolone in the treatment of bullous pemphigoid. AB - BACKGROUND AND DESIGN: Bullous pemphigoid is usually treated with systemic corticosteroids. Side effects are common in elderly patients, justifying the search for adjuvant therapy. This randomized, multicentric unblind study was designed to assess the efficacy of azathioprine or plasma exchange when added to conventional doses of prednisolone. One hundred patients with active disease entered the study. They were randomly allocated to receive 28 days of treatment with oral prednisolone sodium metasulfobenzoate (1 mg/kg per day) either alone or in combination with oral azathioprine (100 to 150 mg/d) or four large-volume plasma exchanges. After 28 days, the prednisolone doses were progressively decreased according to the same strict regimen in the three groups (in combination with oral azathioprine in group 2). RESULTS: The clinical results were evaluable in 98 of the 100 patients included in the study. There was no appreciable difference in the percentages of complete remission of the disease in the three therapeutic groups at 28 days (71%, 80%, and 71%, respectively) or at 6 months (42%, 39%, and 29%, respectively). Severe complications were more often observed among patients receiving azathioprine. At 6 months, 14 of 98 patients had died, without any differences noted among the three study groups. CONCLUSIONS: We conclude that neither azathioprine nor plasma exchange is effective enough to be used routinely as an adjuvant to corticosteroids in the management of bullous pemphigoid. PMID- 8420492 TI - Diagnosis and treatment of nevomelanocytic lesions of the skin. A community-based study. AB - BACKGROUND AND DESIGN: Nevomelanocytic lesions of the skin are frequently removed. However, little data are available concerning the diagnostic accuracy, histologic features, and methods of removal for these common lesions. We identified 2935 consecutive nevomelanocytic lesions examined at four pathology laboratories to determine the accuracy of the preoperative diagnosis, the characteristics of the patients treated, and the methods used for removal. RESULTS: Of 2935 melanocytic lesions, 2087 (71%) were benign nevi, 684 (22%) were dysplastic nevi, 72 (3%) were blue nevi, and 76 (3%) were lentigines. Lentiginous melanocytic hyperplasias with cytologic atypia (20) and melanoma (26) each represented less than 1% of such lesions. Diagnostic accuracy varied by the pathologic diagnosis and the specialty of the treating physician. Dermatologists and plastic surgeons had the highest diagnostic accuracy. Techniques used to remove melanocytic lesions varied by physician specialty and type of lesion. CONCLUSIONS: Despite the high frequency of removal, the methods used to remove nevomelanocytic lesions vary widely. Improving the clinician's ability to distinguish type of lesion may improve care. Studies that determine optimal means for their removal are warranted. PMID- 8420493 TI - Demonstration of silicon in sites of connective-tissue disease in patients with silicone-gel breast implants. AB - BACKGROUND AND DESIGN: Silica, Silastic, and silicone (any organic compound in which silicon replaces carbon) have been associated with a number of connective tissue diseases, most commonly systemic sclerosis (scleroderma). Silicone is known to leak from breast implants and spread to surrounding tissues, including lymph nodes, but silicone's role in the origin and pathogenesis of the inflammation and fibrosis related to such conditions remains controversial. Synovial tissue, alveolar macrophages, and skin, each from three different patients with silicone-gel implants, plus the breast implant capsules from each of the three patients, were examined by light microscopy, transmission electron microscopy, and electron probe microanalysis for the presence of silicon containing material. RESULTS: Silicon was identified within the fibrous breast capsule of each case, associated with a chronic inflammatory cell infiltrate. Silicon was also identified within tissues involved by chronic inflammation and fibrosis, namely, synovium, skin, and alveolar macrophages, in association with clinical, serologic, and histologic evidence of connective tissue disease. All three patients improved after removal of the silicone-gel breast implants. CONCLUSIONS: The presence of silicon-containing material within sites of connective-tissue disease supports a role for silicon in the origin or pathogenesis of such conditions in patients with silicone-gel breast implants. All patients with connective-tissue disease should be questioned about exposure to various forms of silicon. In those patients with known exposure, tissue specimens should be examined carefully for silicon-containing material and, if found, the source should be removed. PMID- 8420494 TI - Pemphigus foliaceus in young women. An endemic focus in the Sousse area of Tunisia. AB - BACKGROUND AND DESIGN: With the exception of some areas in Brazil, pemphigus is an exceptional disease. Several previous observations suggested that pemphigus foliaceus occurred more frequently than expected in Tunisia. To confirm these observations, a 15-month prospective survey was undertaken at the University Hospital of Sousse in Tunisia. RESULTS: During the survey, 23 patients with pemphigus were observed. Twenty of 23 cases were of pemphigus foliaceus that affected only young women. Seven patients had the clinical features of pemphigus herpetiformis. The estimated incidence of pemphigus foliaceus in the Sousse area was four new cases per million inhabitants per year, far above European or North American incidence but lower than the incidence in the most severely affected areas in Brazil. The high prevalence of the herpetiform clinical variant, the young age, the feminine exclusivity, and the absence of familial cases differentiated this Tunisian pemphigus both from pemphigus foliaceus observed in Europe and North America and from the Brazilian fogo selvagem. No etiologic factor was identified. CONCLUSIONS: This study suggests the existence in Tunisia (and possibly in all North Africa) of an endemic form of pemphigus foliaceus restricted to young women. PMID- 8420495 TI - Quantification of chemotactic peptides (C5a anaphylatoxin and IL-8) in psoriatic lesional skin. AB - BACKGROUND AND DESIGN: Psoriatic scale extracts contain a unique chemotactic peptide fraction that is likely to be involved in the induction of rhythmic transepidermal leukocyte chemotaxis. Recent studies have identified the presence of two unrelated chemotactic peptides in this fraction, ie, C5a/C5a des Arg and interleukin 8 (IL-8), and its related cytokines. To investigate their relative contribution to the transepidermal leukocyte migration as well as their interrelationship in psoriatic lesions, we have quantified concentrations of immunoreactive C5a/C5a des Arg and IL-8 in psoriatic lesional scale extracts and those from related sterile pustular dermatoses such as subcorneal pustular dermatosis and pustulosis palmaris et plantaris. RESULTS: The concentrations of C5a/C5a des Arg and IL-8 were more significantly increased in the horny-tissue extracts from lesional skin than in those from noninflammatory orthokeratotic skin (P < .01). The increase of C5a/C5a des Arg concentration was specific to the lesional scale extracts, but showed a rather wide range of variation. By contrast, IL-8 concentration, although consistently increased in the lesional scale extracts, was also moderately increased even in noninflammatory scale extracts prepared from ichthyosis vulgaris. The elevation of IL-8 levels in psoriatic lesions was also confirmed by measuring their levels in cutaneous tissue fluid samples collected from suction blisters. However, unexpectedly, some control samples obtained from normal skin also showed a moderate increase in the IL-8 level. Neutrophil chemotactic activity correlated significantly only with the levels of C5a/C5a des Arg in the scales (P < .05). No such significant correlation was found between chemotactic activity and IL-8 or between C5a/C5a des Arg and IL-8. CONCLUSION: Based on these results, we speculate that, although IL-8 may exert a synergistic effect with C5a/C5a des Arg in the induction of transepidermal leukocyte chemotaxis, it constitutes a proinflammatory cytokine that is involved in the production of the persistent inflammatory changes characterized by a T-lymphocyte infiltration. In contrast, C5a/C5a des Arg seems to be generated only in the inflammatory lesional skin under specific circumstances that preferentially favor complement activation and also seems to play a major role in the induction of cyclic transepidermal leukocyte chemotaxis from "squirting papillae." PMID- 8420496 TI - Lymphomatoid papulosis followed by Hodgkin's lymphoma. Differential response to therapy. AB - BACKGROUND: The association of lymphomatoid papulosis (LyP) with Hodgkin's lymphoma or other lymphomas is well recognized. However, the issue as to whether this represents an independent association or a transformation of one proliferative process to the other remains unresolved. OBSERVATION: A woman with LyP subsequently developed Hodgkin's lymphoma. Combination chemotherapy resulted in apparent cure of the lymphoma but had only a transient effect on the LyP. A literature review revealed a similar difference in response to chemotherapy or radiation therapy in most patients who had LyP and associated lymphoma. CONCLUSIONS: The differential response to therapy in patients with LyP and associated lymphoma suggests that there are biological differences between LyP cells and associated lymphoma cells even though in some patients the immunophenotype and genotype were reported to be identical. However, alternative explanations are possible. In this article we also review studies on other cases of LyP associated with Hodgkin's lymphoma. PMID- 8420497 TI - Clinical classification of cases of toxic epidermal necrolysis, Stevens-Johnson syndrome, and erythema multiforme. AB - BACKGROUND AND DESIGN: To conduct a prospective case-control study about causative factors of severe bullous erythema multiforme, Stevens-Johnson syndrome, and toxic epidermal necrolysis, we needed to define criteria for classifying the cases and standardize the collection of data so that cases could be reliably diagnosed according to this classification. Based on review of case histories and photographs of patients, a group of experts proposed a classification based on the pattern of erythema multiforme-like lesions (categorized as typical targets, raised or flat atypical targets, and purpuric macules) and on the extent of epidermal detachment. An atlas illustrating this classification that included photographs and schematic drawings was developed. We compared the evaluations of 28 cases by four nonphysicians relying on the atlas with the evaluations of the same cases by five experts not using the atlas to determine the usefulness of this atlas for classifying cases according to our nosologic schema. RESULTS: The following consensus classification in five categories was proposed: bullous erythema multiforme, detachment below 10% of the body surface area plus localized "typical targets" or "raised atypical targets"; Stevens-Johnson syndrome, detachment below 10% of the body surface area plus widespread erythematous or purpuric macules or flat atypical targets; overlap Stevens-Johnson syndrome-toxic epidermal necrolysis, detachment between 10% and 30% of the body surface area plus widespread purpuric macules or flat atypical targets; toxic epidermal necrolysis with spots, detachment above 30% of the body surface area plus widespread purpuric macules or flat atypical targets; and toxic epidermal necrolysis without spots, detachment above 10% of the body surface area with large epidermal sheets and without any purpuric macule or target. Using the atlas, the nonexperts showed excellent agreement with the experts. CONCLUSION: This study suggests that an illustrated atlas is a useful tool for standardizing the diagnosis of acute severe bullous disorders that are attributed to drugs or infectious agents. Whether the five categories proposed represent distinct etiopathologic entities will require further epidemiologic and laboratory investigations. PMID- 8420498 TI - Silicone-associated connective-tissue disease. The debate rages. PMID- 8420499 TI - Preparation of human sperm for assisted conception: a comparative study. AB - An ideal sperm preparation technique for assisted conception requires the capacity of accumulating in a relatively small volume the largest number of morphologically normal, viable sperm with good motility. Additionally, this extract of the ejaculate must be free of seminal plasma, leukocytes, bacteria, and other debris. In a comparison of the migration-sedimentation, SpermPrep, and swim-up techniques for sperm preparation, MS proved best at improving sperm motility and morphology in samples from both fertile and subfertile patients. In the fertile group, motility improved by 32% and morphology by 39%, while the subfertile group demonstrated improvements of 49% and 29%, respectively. SpermPrep produced the highest sperm yield after only 15 min of processing. The yield was in the region of 37% for the fertile group and 17% for the subfertile group. All samples demonstrated an improvement in the grade of progressive sperm motility and a decrease in debris content after processing with all three methods. PMID- 8420500 TI - Transferrin concentration in seminal plasma with special reference to serum hormone levels in infertile men. AB - The concentration of transferrin in seminal plasma and serum levels of LH, FSH, testosterone, and prolactin were measured in 94 male patients attending our infertility clinic. The concentration of transferrin in patients with azoospermia was lowest among azoospermia, oligozoospermia, and normozoospermia groups. There was significant positive correlation between the concentration of transferrin in seminal plasma and sperm density. However, seminal plasma transferrin was inversely correlated with serum levels of LH, FSH, and prolactin, respectively. It is suggested that transferrin in seminal plasma contributes to the activation of seminiferous tubules and can be used as a reliable index of Sertoli cell function. PMID- 8420501 TI - Determination of soluble interleukin-2 receptor in human seminal plasma. AB - Following lymphocyte activation a soluble form of TAC antigen is released. This soluble molecule (soluble interleukin-2 receptor, SIL-2R) seems to corresponds to a truncated extracellular part of the membrane-bound TAC antigen, being smaller than its cellular counterpart. SIL-2R was determined and correlated with PMN elastase levels in the seminal plasma of 79 adult men having different concentrations of PMN elastase (0-700 micrograms/L). The normal level of SIL-2R was detected in the seminal plasma of 15 healthy men. The mean +/- SEM was 101 +/ 29 units/mL. The relation between PMN elastase in human seminal plasma as an index for the state of lymphocyte activity and the sperm motility is also investigated. PMID- 8420502 TI - Swim-down: a rapid and easy method to select motile spermatozoa. AB - A method is developed to improve the quality and the number of sperm recovered after laboratory processing, consisting of a swim-down technique. This study was designed to compare quality sperm after selection with a swim-up and a swim-down method. Fifty-nine men, 23-52 years of age, with a period of infertility of 1 to 14 years were included in the study. The efficacy of the swim-down technique as compared to the swim-up technique was assessed by measuring percentage of sperm recovery, sperm motility, the hypoosmotic swelling test, acrosome reaction, and the sperm penetration to zona-free hamster oocyte assay. The swim-down procedure using pure serum as culture medium showed a higher percentage (p < .02) of spermatozoa with forward progression and a higher number of recovered (p < .05) spermatozoa than the swim-up technique. The fertilizability was similar with both methods. The swim-down procedure using pure serum could be useful for intrauterine insemination where a higher number of spermatozoa is required. PMID- 8420503 TI - Semen characteristics of tobacco users in India. AB - Qualitative analysis of semen samples have been compared between 79 different types of tobacco addicts (smokers, chewers, and multiple addicts) with 21 nonaddicts (never consumed any form of tobacco). The percentage of motile sperm and total sperm count of the tobacco chewers are significantly low (p < .05). The frequency of abnormal sperm is also significantly high (p < .001) for smoking and multiple addict groups. Differential effects of smoking and chewing tobacco on sperm characteristics are discussed. PMID- 8420504 TI - Acrosome-reacted sperm in infertile and fertile men using the triple-stain technique. AB - By means of a triple-stain technique, the acrosome reaction in infertile and fertile males was investigated. The highest correlations with the percentage of acrosome-reacted sperm were found for motile sperm concentration (r = .60, p < .01) and percent motility (r = .58, p < .05). Significant correlations were also demonstrated between acrosome reactivity and both sperm concentration (r = .50, p < .05) and velocity (r = .40, p < .05). There was only a weak correlation between the percentage of acrosome-reacted sperm and that of swollen sperm. The ratio of acrosome-reacted sperm demonstrated in oligoasthenozoospermic patients was significantly lower than that observed in normozoospermic or fertile males (both, p < .05). There was no significant difference in the ratio of acrosome-reacted sperm between normozoospermic and fertile males. However, all the cases in the latter group had 15% or more acrosome-reacted sperm, whereas the parameter was widely distributed in the former group and some in this group had a decreased ratio. It would appear that a disturbance of the acrosome reaction is one of the causes of reduced fertility potential in normozoospermic males. PMID- 8420505 TI - Semen analysis in insulin-dependent/non-insulin-dependent diabetic men with/without neuropathy. AB - The present study deals with the diabetic neuropathies prevailing in the male population. In this investigation 100 insulin-dependent diabetes mellitus (IDDM) and 314 non-insulin-dependent diabetes mellitus (NIDDM) patients with and without an objective evidence of neuropathy, having an age span of 15 to 80 years and the duration of diabetes distributed over 1-33 years were included along with age matched nondiabetic controls. The diabetic subjects were evaluated for semen analysis. Results of semen analysis showed a highly significant increase (p > .0005) in total sperm output and sperm concentration in both IDDM and NIDDM neuropathic diabetic men. On the other hand, sperm motility and semen volume were found to be about 30 and 60% less, respectively, in IDDM and NIDDM patients, where as sperm morphology and quality of sperm motility remained unaffected. A comparison between IDDM and NIDDM neuropathic and non-neuropathic diabetic groups further indicated a nonsignificant difference in the parameters of semen analysis, thus suggesting an endocrine basis for the sexual disturbances of diabetic neuropathy. A significant rise in total sperm output in both IDDM and NIDDM neuropathic diabetic patients and a significant decrease in semen volume in both types of diabetic patients thus suggests some kind of Leydig cell hyperplasia, which in turn may stimulate spermatogenesis and atonia of the bladder and urethra, resulting in retrograde ejaculation. PMID- 8420506 TI - Hyperviscosity and hypofunction of the seminal vesicles. AB - The study was designed to determine whether hyperviscosity of the semen sample is related to dysfunction of the male accessory glands. It was carried out on men who consecutively attended an infertility clinic between June 1989 and June 1991, and the men were grouped according to viscosity of semen samples (normal viscosity or higher viscosity). Semen samples from 229 infertility patients were studied. From these, 155 had normal viscosity and 74 showed hyperviscosity. The effect of hyperviscosity of semen samples on seminal quality and the function of the prostate was evaluated by acid phosphatase measurement, and the seminal vesicles by measurement of corrected fructose. Sperm motility (grades II-III), sperm vitality, and corrected fructose were significantly reduced in samples with high viscosity (p < .05). A high prevalence of hyperviscosity in semen samples was associated with only hypofunction of the seminal vesicles. In fact, 36.5% of subjects with hyperviscosity showed reduced levels of corrected fructose. The same association with hyperviscosity was not observed when only hypofunction of the prostate was present, or when hypofunction of both prostate and seminal vesicles was present (P:NS). Further analysis showed that high viscosity is observed mainly when corrected seminal fructose levels were below 1.5 mg/mL x 10(6) spz/mL. It would appear that hyperviscosity affects sperm motility and is associated with hypofunction of the seminal vesicles. PMID- 8420507 TI - Differential sperm motility scoring and sperm ATP concentrations. AB - Sperm motility is one of the most important parameters in semen analysis. In this study the motility scoring method of Hellinga was used. Adenosine triphosphate (ATP) is the main energy source used by the sperm flagellum to initiate and propagate forward motility. This study was conducted to evaluate the interrelationships between sperm ATP and differential motility. The differential motility method supplemented the conventional by which the quantitative and qualitative motility were estimated. There was a good correlation between the differential motility scoring method and sperm [ATP], although the latter did not necessarily improve the diagnostic capacity. PMID- 8420508 TI - Energy metabolism of spermatozoa during pronucleus formation induced in vitro by heparin-reduced glutathione. I. Glucose uptake. AB - Glycolitic metabolism under basal conditions and its modifications by the combined action of heparin and GSH were studied in human sperm. Respirometric data indicated that the amount of U. L. [14C]-glucose converted to 14CO2 increased with the incubation time, being almost linear for up to 60 min and then leveling off at 120 and 150 min (594 and 620 nmol of [14C]-glucose/10(8) spermatozoa, respectively). When spermatozoa were incubated in the presence of heparin-GSH such behavior completely changed, showing a decrease (approximately 50%) in glucose metabolism with values of 254 and 366 nmol of [14C]-glucose/10(8) spermatozoa at the same incubation times as the basal consumption. When these results were compared with the kinetic of the swollen nuclei it was seen that at 30 min 44% of the spermatozoa have its nuclei swollen with a glucose uptake value of 91 nmol/10(8) spermatozoa, and at 150 min when nearly all the spermatozoa nuclei are swollen (95%) the glucose uptake increases fourfold more than the initial rate at 30 min. Therefore, it is possible to suggest the existence of an energy contribution by the sperm to the male pronuclei formation mechanism. PMID- 8420509 TI - Platform motorized wheelchairs in congregate care centers: a survey of usage and safety. AB - A survey of residents was done in three congregate care facilities to determine the prevalence and safety of platform motorized wheelchairs (PMWs) otherwise known as three-wheeled motorized wheelchairs or scooters. Thirty-one drivers of these vehicles were identified, 30 of whom consented to be interviewed. The mean age of the drivers was 80.7 years and 84% were women. Arthritis was the most frequent reason for use of the PMW (47%), followed by neurologic problems (23%) and orthopedic problems (23%). Of this group, 77% had their PMW prescribed by a physician, most commonly an orthopedic surgeon (40%) with only 14% being prescribed by a geriatrician or physiatrist. Thirty percent of the drivers felt other drivers in their facility drove unsafely. Most residents (57%) had no training in the use of the vehicle and two admitted they had a medical condition that made it difficult to drive. Five accidents were reported, two of which involved either a serious injury or property damage. Based on the results obtained from this survey, we conclude that driver safety evaluation should be done on all users of PMWs in congregate care facilities. PMID- 8420510 TI - A dynamic pronation orthosis for the C6 tetraplegic arm. AB - Tetraplegic arms of patients with spinal cord injury at C5 or C6 level are often held in a flexed supinated position. In supination the function of the hand is limited. This report presents a newly developed orthosis giving active pronation of the forearm for a C6 complete tetraplegic. The orthosis consists of a pretightened coiled spring that gives a torque around the longitudinal axis of the forearm. The distal part of the spring is connected to a cuff around the wrist, and the proximal part to two fitting planes on the upper arm. With the orthosis, a 27-year-old patient with a C5-C6 spinal cord injury was able to participate in activities of daily living and to perform table top activities. This new orthosis provides pronation and markedly increases the patient's degree of independence. PMID- 8420511 TI - Functional status measures and reimbursement. PMID- 8420512 TI - Krusen Award to Dr. Opitz. PMID- 8420513 TI - Pin sensation as a predictor of extensor carpi radialis recovery in spinal cord injury. AB - The purpose of this prospective study was to determine if initial preservation of C5 pin sensation could predict extensor carpi radialis (ECR) motor recovery in the zone of partial preservation after spinal cord injury (SCI). The study compared initial C5 pin sensation and initial ECR strength as predictors of ECR motor recovery after C4 and C5 SCI. The initial motor and sensory evaluation was done less than seven days after injury. Motor power was graded from absent (0/5) to normal strength (5/5). Pin sensation was graded as absent (0/2) decreased (1/2) or normal (2/2). The manual muscle test was performed weekly to four weeks and than at three, six, and 12 months after injury. An ECR muscle grade of > or = 3/5 was defined as recovery. C5 pin sensation at the lateral antecubital fossa significantly correlated with ECR recovery to > or = 3/5 (p < 0.001). Only two of nine subjects with absent C5 pin sensation recovered ECR to > or = 3/5, whereas 14 of 15 subjects with 1/2 or 2/2 C5 pin sensation had ECR motor recovery to > or = 3/5. All ten subjects with 1/5 or 2/5 initial ECR strength had ECR motor recovery. Five of six subjects with 0/5 initial ECR strength and subsequent ECR recovery to > or = 3/5 were found to have C5 pin preservation. In conclusion the presence of 1/5 to 2/5 initial ECR strength or 1/2 or 2/2 C5 pin sensation were highly significant predictors of ECR motor recovery to > or = 3/5 in C4 and C5 motor complete quadriplegic subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420514 TI - Kinetic analysis of dynamic transitions in stance support accompanying voluntary leg flexion movements in hemiparetic adults. AB - The purpose of this study was to examine the extent to which adult hemiparesis due to cerebrovascular accident may alter the dynamic transitions from bipedal to single limb stance that accompany rapid voluntary single leg flexion movements while standing. Eight postacute hemiparetic adults performed rapid single-leg flexion movements with the paretic (PL) and nonparetic (NL) limbs while standing on two separate force platforms that recorded the individual and resultant lateral horizontal (FY) ground reaction force (GRF) components acting on the body in the frontal plane. The results showed that for NL movements, the ipsilateral FY force-time integral contributed a significantly greater proportion to the resultant FY than that recorded beneath the upcoming stance PL, whereas PL movements showed a reverse trend indicating differences in the spatial distribution of GRFs. For some subjects, delays in the initial FY onset times between limbs and reversals in the normal direction of initial force application beneath the PL were observed. Such changes in the spatial and temporal aspects of GRF production may affect dynamic lateral weight transfer function regardless of the direction of total body motion. Implications for clinical practice pertaining to interventions that emphasize speed as well as magnitude of paretic muscle torque production, and factors related to the selection of movement activities for the retraining of dynamic weight transfer function are discussed. PMID- 8420515 TI - Health care costs of veterans with multiple sclerosis: implications for the rehabilitation of MS. VA Multiple Sclerosis Rehabilitation Study Group. AB - We retrospectively determined health care costs among veterans with multiple sclerosis (MS) and correlated the costs with neurologic dysfunction. Total health care costs for the 165 patients averaged $35,000/year. VA benefits and homecare together accounted for 85% of the total costs. Total health care costs correlated with two measures of neurologic dysfunction, the Expanded Disability Status Scale (EDSS) (r = 0.61, p < 0.001) and the Incapacity Status Scale (ISS) (r = 0.64, p < 0.001). The costs of VA benefits, homecare, and hospitalizations also correlated with the EDSS, ISS, and other measures of neurologic dysfunction whereas the cost of outpatient clinic visits did not. In a period of three years, there were 40 hospitalizations, at a total cost of $412,800, that were potentially preventable with appropriate outpatient management. Improving selfcare and avoiding preventable hospitalizations might lower the considerable health care costs of MS. PMID- 8420516 TI - Prospective evaluation of the course of disc herniations in patients with proven radiculopathy. AB - Although surgery is often recommended as the definitive treatment for radiculopathy when definite disc herniation is demonstrated with imaging techniques, complete improvement can occur with nonoperative treatment. However, what happens to the disc in the latter circumstance is not well defined. We report the first prospective study in subjects with proven radiculopathy and definite disc herniation who improve with nonoperative management to determine what occurs to the herniated disc material. Eighteen subjects with lower extremity pain or paresthesia, positive straight leg raising, weakness in a myotomal distribution, reflex asymmetry, or electromyogram evidence of radiculopathy were studied. Subjects were admitted to the study if computed tomography (CT) scanning demonstrated definite disc herniation corresponding to the side and level of the radiculopathy. After complete clinical improvement, repeat CT scan was performed at six to 18 months after the initial study. The CT scans were interpreted separately by two neuroradiologists. Disc herniations were characterized by size (large, moderate, or minimal); the presence of absence of free fragments; and location. Follow-up scans were compared with the original study and characterized as resolved, improved, or unchanged. Fourteen subjects completed the study, an additional three had operative treatment, and one refused repeat scanning. Subjects were followed an average of 30.4 months with no recurrence of radicular symptoms during this follow-up period in 13 patients. One had recurrence of symptoms at 21 months and surgery at 26 months. Six follow-up scans (43%) were interpreted as completely resolved, five (36%) as improved, and three (21%) as unchanged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420517 TI - Interrater reliability of the Cybex EDI-320 and fluid goniometer in normals and patients with low back pain. AB - We compared the interrater reliability of the Cybex Electronic Digital Inclinometer-320a (EDI) and the fluid goniometer for measuring lumbar spine range of motion (ROM) in flexion, extension, and prone extension in adults not experiencing back pain and low-back pain patients. We also investigated whether prior palpation training improves reliability of lumbar ROM measurements using these tools. A repeated measures factorial design was used with a 6 x 6 Latin square to balance the order of testing. Twelve adults not experiencing back pain and six subacute low-back-pain patients were measured by two trained and one untrained therapist in all positions with both tools. The L1 and S2 spinous processes were palpated and marked for each measurement. Results of several three way ANOVAs indicate that no significant differences exist between the tools for nonpatients or patients regardless of position. Using the Intraclass Correlation Coefficient both tools exhibited acceptable reliability under most conditions. The fluid goniometer exhibited better reliability for patients than nonpatients, although this pattern was not present with the EDI. With the exception of extension, prior palpation training did not improve measurement with either tool. PMID- 8420518 TI - Assessment of patient function after limb-sparing surgery. AB - Cancer rehabilitation is becoming more of a focus for the field of physiatry due to increased longevity and the side effects of treatment. In order to investigate the rehabilitation needs of patients undergoing limb-sparing procedures, chart analysis was conducted on 17 children treated for primary bone tumors by resection and an expandable endoprosthetic replacement. Each patient underwent a course of postoperative inpatient and outpatient physical therapy and was followed over an average of 2.5 years. Gait training was relatively straightforward and in seven patients required neither orthosis nor ambulatory aid. The other ten patients walked with a knee orthosis, axillary crutches, or both. Until the time came for reoperation to lengthen the implant, a shoe lift of 1in maximum was added to compensate for the limb length discrepancy. These findings compare favorably with the more complex requirements of high proximal amputees with external prostheses, including more difficult gait training and the need for frequent adjustments, as well as prosthetic replacement as the children grow. It is clear that children undergoing limb-sparing surgery have special needs that should be addressed, including early mobilization, gait training, adjustment to repeated brief hospitalizations for lengthening, and continued follow-up to monitor their activity restriction. PMID- 8420519 TI - Median-radial latency difference: its use in screening for carpal tunnel syndrome in twenty patients with demyelinating peripheral neuropathy. AB - The median-radian latency difference (MRLD) has been advocated as a screen for mild carpal tunnel syndrome (CTS); however, its validity in the face of even mild peripheral neuropathy is questionable. The MRLD of 20 consecutive individual referred with electrodiagnostic evidence of demyelinating peripheral neuropathy was obtained using standard techniques. The MRLD was statistically prolonged in 85% of the subjects without corroborating evidence of CTS (p < .05). Additionally, statistically significant (p < .05) differences were noted between the percentage of prolonged latencies of the radial sensory (50%) and the median sensory (100%) and all sensory nerves (97%). This investigation suggests that superficial radial nerve involvement in demyelinating peripheral neuropathy occurs later than other sensory nerves. This data indicates that though the MRLD may serve as a sensitive screening method for CTS, it has a low specificity therefore, a more extensive evaluation for mild peripheral neuropathy is always indicated. PMID- 8420520 TI - Effects of TENS and topical skin anesthesia on soleus H-reflex and the concomitant influence of skin/muscle temperature. AB - The purpose of this study was to determine, in ten healthy subjects, the extent of soleus motoneuronal excitability during conditions of increased (transcutaneous electrical nerve stimulation [TENS]), decreased (Xylocaine [lidocaine]a anaesthesia) and normal (placebo anaesthesia) cutaneous inputs. Increased cutaneous activity was evoked using a TENS unit, with the two pairs of electrodes placed respectively over the Achilles (S2 dermatome) and tibialis anterior (L5 dermatome) tendons. Experimental and placebo topical anaesthesia were obtained after rubbing Xylocaine (5%) and Vaselineb ointment, respectively, on the skin surface overlying the Achilles tendon. Sets of ten H-responses (Hmax/2) were evoked at a frequency of 1 shock/30s and averaged at regular time intervals before, during and after the testing conditions. The results showed a gradual increase (up to 40% after 20 minutes) of H-reflex amplitude during TENS regardless of whether it was applied on the L5 or S2 dermatome. Furthermore, placebo anesthesia (Vaseline) caused the same gradual facilitatory response (up to 100% after 50 minutes) as that obtained during Xylocaine anaesthesia. Power spectral analysis of the H-responses obtained over time showed that the increase in the peak-to-peak H-response value was accompanied by a shift of the spectral content toward low frequencies. This shift occurred concomitantly with a cooling of the skin overlying the soleus muscle. PMID- 8420521 TI - Conventional and acupuncture-like transcutaneous electrical nerve stimulation excite similar afferent fibers. AB - The purpose of our study was to determine whether similar or different peripheral afferent fiber(s) is(are) activated by "conventional" transcutaneous electrical nerve stimulation (TENS) applied at low intensity-high frequency, as opposed to "acupuncture-like" TENS administered at high intensity-low frequency. The electrical stimulation was delivered to the median nerve at the wrist of 17 healthy subjects. For conventional TENS, single pulses were applied at an intensity of 3 X T (sensory threshold). Two kinds of acupuncture-like TENS were studied: single pulses at 0.1Hz, and trains of 100Hz pulses at 4Hz, both delivered at an intensity greater than 3 X T. Thirty compound action potentials per type of stimulation were recorded over the median nerve in the cubital fossa and averaged. The results showed that the mean conduction velocities of the afferent fibers excited by conventional TENS, single pulse, and short-train acupuncture-like TENS ranged from 50.3 to 65.4, 50.0 to 63.5, and 41.3 to 54.8m/s, respectively. Thus, conventional and acupuncture-like TENS activated similar fiber types, predominantly in the A alpha beta range. Our findings suggested that the effects of these two types of TENS may be mediated by the activation of similar peripheral afferent fibers. PMID- 8420522 TI - Traumatic brain injury and chronic pain: differential types and rates by head injury severity. AB - Traumatic brain injury has been associated with many physical and neurobehavioral consequences, including pain problems. Documented most has been the presence of posttraumatic headaches that are associated with the postconcussion syndrome. This study therefore examined types and rates of chronic pain problems in patients seen in an outpatient brain injury rehabilitation program. A total of 104 patients were evaluated, 66 of whom were male and 38 female, and the average time postinjury was 26 months. Headaches were the most frequent chronic pain problem across both mild and the moderate/severe groups, although in the former, a significantly higher frequency was noted (89%) when compared against the latter group. The same relative rates were seen for chronic neck/shoulder, back, and other pain problems. The mild group also showed a higher frequency of concurrent pain problems, whereas in the moderate/severe group only one patient had more than one chronic pain problem. Results also showed that in the mild group neck/shoulder accompanied headaches 47% of the time, and back pain coexisted with headaches 44% of the time. These results underscore the high frequency of chronic pain problems in the mild head injury population and implicate the need for avoiding the mislabeling of symptoms such attentional deficits or psychological distress as attributable only to head injury sequelae in those with coexisting chronic pain. Early identification and intervention of pain syndromes in the mild head-injury population is also suggested. PMID- 8420523 TI - Erythropoietin profile in spinal cord injured patients. AB - Patients with spinal cord injury (SCI) have a high incidence of anemia. Intact erythropoietin (EPO) production is essential to the maintenance of erythrocyte mass and prevention and correction of anemia. However, the effect of chronic SCI on EPO production remains unclear. We measured plasma EPO concentration in 83 men with longstanding SCI and a group of normal able-bodied individuals. The SCI patients showed a significant reduction in hematocrit, a high prevalence of anemia, and an increased plasma EPO concentration. Active smokers showed a significantly higher hematocrit and lower EPO concentration than nonsmokers. No significant difference was found in hematocrit or EPO between individuals with paraplegia and those with quadriplegia. A negative correlation was found between EPO and hematocrit in SCI patients lacking significant lung disease. Thus, in the absence of renal insufficiency, EPO response to anemia is qualitatively preserved in SCI patients and is largely independent of the level of injury. PMID- 8420524 TI - Energy storing property of so-called energy-storing prosthetic feet. AB - The purpose of this study is to evaluate the energy storing and releasing property of 14 different prosthetic feet, in which so-called energy storing prosthetic feet are included. The prostheses were fitted to a young male amputee who walked on a level and slope walkway. Gait-analysis data were obtained by an automatic gait measurement system from which the ankle moment and ankle angular velocity were calculated. From these values, the power and the energy at the ankle joint were calculated. The prosthetic feet were ranked according to the total amount of energy, that is the sum of energy being stored and released. Both of the stored and released energy are important feature of the prosthetic feet. According to these evaluation criteria, we ranked the feet into three groups. The high total energy feet were Flex-Walk,a SAFE II,b and High Functionc. The medium total energy feet were SAFE,b Greissinger,d Seattle,e Copy II,f and Dynamic.d The low total energy feet were Seattle Light,e Fore Joint,g Quantum,h SACH,dSTEN,i and Uniaxial.d As for the subjective feeling, our subject preferred higher energy absorbing feet, in which a somewhat high level of energy was stored but low level of energy was released. PMID- 8420525 TI - Effect of "standing" on spasticity, contracture, and osteoporosis in paralyzed males. AB - The effect of "standing" in a frame on spasticity (clinical assessment and H reflex), contracture (lower extremity joint range of motion), and osteoporosis (dual photon absorptiometry) was studied in six paralyzed males (mean age 49 yr) who had been confined to wheelchairs for an average of 19 years. Standing time averaged 144 hours over a mean of 135 days. Clinical Assessment measured reflexes, tone, and clonus in the legs. Results revealed no important differences between initial and final scores for clinical assessment and joint range of motion. In three subjects for whom H-reflexes were found, latency and amplitude were not altered by "standing." Bone density was normal in the lumbar spine but significantly reduced in the femoral neck. "Standing" did not modify the bone density in any site. A follow-up interview revealed that 67% of subjects continued to "stand" and felt healthier because of it. In summary, "standing" had no ill effects, did not alter measured variables, and had a positive psychological impact. PMID- 8420526 TI - Scurvy--a forgotten disease. AB - A four-year-old male child was reported with sudden inability to stand walk of 10 days duration. Acute paralytic poliomyelitis was diagnosed in the presence of fever, respiratory infection, muscle tenderness, and weakness in lower extremity in an unimmunized child. Reevaluation after a week showed swelling, thickening, and tenderness of left femur. Plain x-ray of the left thigh was advised to exclude underlying osteomyelitis of the femur. X-ray showed features of scurvy. The child responded well to parenteral vitamin C therapy and resumed normal walking with in two weeks. Scurvy can present as pseudoparalysis of acute onset without any other signs even in older children. It emphasized the need for an index of suspicion of scurvy in a case of suspected paralysis. Dramatic response to vitamin C therapy can be used to confirm the diagnosis in the absence of laboratory support. Health education can be accelerated to prevent a disease like scurvy. PMID- 8420527 TI - Hyperthermia associated with baclofen withdrawal and increased spasticity. AB - Rapid baclofen withdrawal is known to cause markedly increased spasticity, but high fever associated with this complication has not been reported. We describe a 13-year-old boy with sensory incomplete C1 quadriplegia two years after injury who was on 200mg of baclofen per day for spasticity. Concerns about adverse side effects prompted tapering of his baclofen. Severely increased spasticity was noted with associated hyperthermia to 107 degrees F after the dosage was gradually decreased. Sepsis work-up was negative, head computed tomography scan was unchanged, and electroencephalogram showed no epileptiform activity. Cooling blankets, intravenous diazepam, and return of baclofen to 160mg per day decreased spasticity and normalized body temperature without recurrence of hyperthermia. Possible fever etiology is the hypermetabolic state associated with the acute return of spasticity. PMID- 8420528 TI - A case of cerebral infarction in association with free protein S deficiency and oral contraceptive use. AB - The purpose of this article is to describe protein S deficiency as a cause of cerebral infarction in the young adult. We report a 27-year-old previously healthy woman with a primary free protein S deficiency, who developed a left temporoparietal infarction. Protein S is a naturally occurring vitamin K dependent protein which, in conjunction with active protein C, inhibits the clotting cascade. Protein S deficiency is known to be of clinical significance in patients with deep venous thrombosis or pulmonary emboli and in these patients, treatment is long-term anticoagulation. Protein S deficiency has been found to be associated with cerebrovascular occlusion and may possibly warrant long-term anticoagulation in these patients as well. Measurement of total and free protein S levels should be part of the evaluation for any young adult who has had a stroke. PMID- 8420529 TI - Velocity change and fatality risk in a crash--a rule of thumb. PMID- 8420530 TI - Guidelines for the development of safety-related standards for consumer products. AB - Product safety standards, issued by official standardization bodies, play an important part in consumer protection policy. Many of these standards exhibit considerable shortcomings, which limit their applicability and effectiveness. This paper deals with a number of problems in the development of safety standards, in particular for consumer products. Recently published guidelines for safety-related standards are discussed, and additional recommendations are developed. In addition to the traditional problems in standardization, attention is devoted to product information and the generalizability of test results. The analysis and recommendations in this paper do not only apply to standards, but also to other documents containing test methods for consumer products, e.g. for comparative testing, certification, and product development. PMID- 8420531 TI - Fatal accidents of older drivers. AB - Fatal accidents of drivers aged 65 or more in Finland in 1984-1989 were compared with those of the statistically safest age group of 26-40 with special emphasis on self-caused accidents. The basic material consisted of 769 multidisciplinary investigated traffic accidents. Older drivers had an overall responsibility ratio [(single + guilty)/total] of .89 versus .61 for the comparison group; in collisions between vehicles this ratio was .87 versus .50. The number of accidents per driver's license increased with age in old drivers. Accidents caused by older drivers were different from those of the comparison group. Old drivers typically collided in an intersection with a crossing vehicle, which they did not notice at all, or saw so late that they did not have enough time to try an avoiding maneuver. Accident characteristics and their implications for safety research and countermeasures are discussed. PMID- 8420532 TI - Mechanism of injury from air bag deployment loads. AB - Loadings induced by deploying currently representative air bags were studied with driver surrogates (anesthetized swine) leaning against the system during inflation. Torso injury mechanisms were studied in a physiologic model, supported against a static steering wheel-mounted air bag system. Severe and extensive chest and abdominal injuries to the swine were observed in the tests. Loading caused by air bag deployment can occur in either of two phases. The first phase represents the initial punch out of the bag from the module; the second phase represents the membrane force of the inflating bag. Statistical analysis indicated that punch out induced injury because of the high rate of loading to the surrogate body region in direct contact with the air bag module. Membrane forces induced injury by high compression over a larger area. Punch-out loading might be reduced by allowing the bag to escape from other parts of the container not in contact with the driver during deployment. Loading by the inflating bag might be reduced by using a compliant steering system to support the module. The amount and rate of generated gas had only marginal effect on the cumulative injury. Even an inflator with inadequate gas output to protect a properly seated occupant had sufficient energy to induce severe injuries in a surrogate in contact with the inflating module. Analysis of the field relevance of the results must consider not only the injury potential given that a driver is in direct contact with the air bag module at the time of deployment, but also the expected field frequency of such an event. Analysis of the field relevance of the results must also consider the correlation of the laboratory test environment with real world exposure. PMID- 8420533 TI - The potential of community diagnosis as a tool in planning an intervention programme aimed at preventing injuries. AB - Injuries due to accidents are a serious public health problem in Sweden as in the rest of the world. In Sweden injuries are the most frequent cause of death among people under the age of 50. More than 75% of all injuries occur in the home or surrounding area. Most accidents strike children, teenagers, and the elderly. Many accidents can be avoided. Prevention is therefore important. A community intervention programme for the prevention of accidents has been developed in the municipality of Sollentuna in Stockholm County. During the planning phase, a basic analysis of the local community was found to be useful, i.e. a Community Diagnosis, which includes three stages: description, analysis, and a health action programme. This report concentrates on the first two stages. To make a community diagnosis, some basic data are needed. In this report the relevance of the existing registers to the Community Diagnosis model is discussed. It is also shown how the Community Diagnosis model helped in the planning phase: the community profile demonstrated whom the prevention should be aimed at, the health profile emphasized the importance of accident prevention, the health risk profile showed where to change the environment, and last, the organizational profile elucidated how preventive work should be organized. PMID- 8420534 TI - Elderly drivers in Germany--fitness and driving behavior. AB - During the period between 1985 and 2000 the number of elderly people (65 years and older) holding a driver's licence will probably double in Germany. Under the broadly accepted assumption that the elderly drive less safely than other age groups, it is suspected that this will negatively affect traffic safety. The central topic of the study concerns the results of driving tests with 80 elderly drivers (60-82 years old), compared with a reference group of 30 middle-aged motorists (40-50 years), and their relation to laboratory performance data. In the laboratory marked differences were found concerning visual acuity by daylight (even when deficiencies were corrected by visual aids) and in the dark, performance in a traffic-related tachistoscopic perception test, and notably in the amount of time needed in tracking and reaction tests. The performance of elderly drivers proved worse in all of these laboratory tasks. On the other hand, in driving tests in the overwhelming number of traffic situations the elderly did not differ unfavorably from the middle-aged drivers. Possible explanations for these findings are considered. PMID- 8420535 TI - Some contagion models of speeding. AB - Drivers' decision on whether or not to speed are only partially predicted by attitudes towards speeding, beliefs about the consequences of speeding, and police efforts to enforce speed restrictions. We propose that a significant role may be played by drivers' comparisons of their own speed with that of other, nearby drivers. Such comparisons may lead to self-amplifying, nonintuitive consequences at the aggregate level. We present several simple models of these social contagion processes and demonstrate analytical strategies for tracing their implications. We also present some preliminary data suggesting that significant contagion effects exist. Finally, we outline some promising directions for research on contagion effects, and trace their implications for enforcement efforts. PMID- 8420536 TI - Safety restraint usage in fatal motor vehicle crashes. AB - In 1987, following implementation of the Washington safety restraint mandatory usage law, collision statistics indicated an increase in motor vehicle crashes in which occupants had died while utilizing safety restraints. Because of concerns expressed by restraint use law advocates, a "Fatal Collision Research Team" was established to evaluate and reconstruct collisions in which restrained occupants had died. Collisions were analyzed over a three-year period and included 337 crashes involving 1,058 occupants, of whom 446 died. The team analyzed fatal collisions involving restrained occupants and documented restraint usage as well as severity of the crash. Crash severity was measured using either the Collision Deformation Classification (CDC) system, which defines the extent of crush to the vehicle, and/or intrusion into the passenger occupant area, or by measuring deceleration forces (delta v) acting on the vehicle, where sufficient data existed to do so. The survivability of the crash was determined individually for each occupant and independently of whether the occupant lived or died, as well as whether the occupant was restrained or not. The major findings were: (i) that in the majority of cases where a restrained occupant died, the fatality can be attributed to the extent of vehicle crush and deceleration forces, i.e. the crash forces and dynamics made the crash nonsurvivable, and (ii) restraint misuse seemed not to be a major contributory factor in fatal crash injuries. PMID- 8420537 TI - Male/female driver characteristics and accident risk: some new evidence. AB - Due to the high correlation that generally exists among driver characteristics, as well as the overall complexity of the factors involved, the role that gender plays in the relationship between driver characteristics and accident risk has been difficult to quantify using traditional statistical approaches. This paper attempts to provide some new insight by using hazard functions and a sample of University of Washington drivers. The subsequent empirical analysis uncovers important differences in the relationship between male and female driver characteristics and their respective accident risk. PMID- 8420538 TI - A new method for making interstate comparisons of highway fatality rates. AB - Highway fatality rates vary significantly among jurisdictions. Before attributing these variations to policy differences, it is important to account for exogenous factors (e.g. weather) that are beyond the control of policy makers. A new method is developed to identify states that are doing better or worse than expected. The method is applied to the 48 contiguous American states using highway fatality data for the years 1975 to 1986. A fixed effects linear model is used to estimate the fatality rate for each state, taking into account exogenous factors. Results indicate that many states have higher or lower fatality rates than is expected based on a ranking of states according to crude fatality rates. Policy implications are discussed. PMID- 8420539 TI - The specific accident factor of older employees. AB - The aim of this study was to find out the specific accident factor of older workers by comparing them to younger workers. The data consist of 99 occupational accidents in which 102 persons were seriously injured. Eighteen of the victims were over 50 years old. The particular accident risk of older employees was getting run over by moving vehicles. In future, work processes will further speed up, and the number of older workers will increase. These two trends are contradictory, and imply an increased accident risk for older employees. PMID- 8420540 TI - Gastrointestinal lymphoma: a surgical perspective. AB - The incidence of gastrointestinal lymphoma, especially gastric lymphoma, has increased threefold since 1960. The stomach is affected most frequently, followed by the small bowel and colon. Gastric lymphoma can be diagnosed using endoscopic biopsies in about 80% of cases. Small bowel and colonic lymphomas are usually diagnosed at operation. The most important factors affecting survival are pathologic stage and nuclear grade. Since about half of gastric lymphomas are low grade, but virtually all small bowel lymphomas are high grade, survival in patients with gastric lymphomas, as a group, is better than that in patients with small bowel lymphomas. Most series have found that surgical resection provides a marked survival advantage. However, no controlled trials have determined the optimal treatment for these patients. In addition to surgical resection, virtually all patients should receive adjuvant chemotherapy and/or radiotherapy. For gastric lymphomas, average 5-year survival rates by stage are about 85% (stage IE), 65% (stage IIE), and 25% (stage IVE). PMID- 8420541 TI - Plans for Medicare outpatient payment reform: Part 1. PMID- 8420542 TI - Hypercalcemia of malignancy: pathophysiology and implications for treatment. AB - As the most common metabolic consequence of cancer, hypercalcemia of malignancy is often encountered in patients with solid tumors, most often lung, head and neck, and breast carcinomas. Since the clinical consequences of hypercalcemia of malignancy may be fatal, an understanding of its pathogenesis and skeletal related factors that may lead to hypercalcemia is important in directing therapy. It is also important to have reasonable expectations and goals outlined before initiating therapy in an individual patient. Interventions aimed specifically at osteoclast inhibition normalize serum calcium levels while treating the final common pathway responsible for the disorder; these include calcitonin, plicamycin, gallium nitrate, and the bisphosphonates. An important consequence of the advent of antiresorptive therapy has been the initiation of clinical trials aimed at preventing skeletal-related morbid events from bone metastases. These trials may ultimately prove to be the most significant benefit of osteoclast inhibitor therapy for patients. PMID- 8420543 TI - Ongoing progress in the treatment of childhood rhabdomyosarcoma. AB - During the past 2 decades, several different multidisciplinary treatment studies of childhood rhabdomyosarcoma have been conducted, some as single-institution protocols and others as multi-institution cooperative endeavors. Much of our current understanding of the natural history and histopathology, as well as response to treatment, is derived from these studies. This understanding has allowed for the continued evolution of treatment, which has already resulted in a dramatic increase in overall survival rates. Current treatment trials are aimed at maintaining high cure rates while reducing treatment-related morbidity, and exploring innovative regimens to improve the outlook of poor-risk patients. The improved outcome in children with rhabdomyosarcoma is a tribute to the multi institutional, multidisciplinary approach to cancer research and treatment. PMID- 8420544 TI - Expanded programme on immunization. Global advisory group--Part I. PMID- 8420545 TI - The stability of the visual world. PMID- 8420546 TI - Motion parallax and other dynamic cues for depth in humans. PMID- 8420547 TI - How insects infer range from visual motion. PMID- 8420548 TI - Subcortical analysis of visual motion: relative motion, figure-ground discrimination and self-induced optic flow. PMID- 8420549 TI - Cortical processing of visual motion. PMID- 8420550 TI - Decoding of retinal image flow in insects. PMID- 8420551 TI - Visual stabilization in arthropods. PMID- 8420552 TI - The sensory-motor link in motion-dependent flight control of flies. PMID- 8420553 TI - Multisensory control in insect oculomotor systems. PMID- 8420554 TI - Motion psychophysics. PMID- 8420555 TI - Detecting visual motion: theory and models. PMID- 8420556 TI - Development of OKN. PMID- 8420557 TI - Subcortical optokinetic mechanisms. PMID- 8420558 TI - The optokinetic response in primates and its possible neuronal substrate. PMID- 8420559 TI - The role of attention and cognitive processes. PMID- 8420560 TI - The sensing of rotational and translational optic flow by the primate optokinetic system. AB - In primates, there are several reflexes that generate eye movements to compensate for the observer's own movements. Two vestibuloocular reflexes compensate selectively for rotational (RVOR) and translational (TVOR) disturbances of the head, receiving their inputs from the semi-circular canals and otolith organs, respectively. Two independent visual tracking systems deal with any residual disturbances of gaze (global optic flow) and are manifest in the two components of the optokinetic response: the indirect or delayed component (OKNd) and the direct or early component (OKNe). I hypothesize that OKNd--like the RVOR--is phylogenetically old, being found in all animals with mobile eyes, and that it evolved as a backup to the RVOR to compensate for residual rotational disturbances of gaze. Indeed, optically induced changes in the gain of the RVOR result in parallel changes in the gain of OKNd, consistent with the idea of shared pathways as well as shared functions. In contrast, OKNe seems to have evolved much more recently in frontal-eyed animals and, I suggest, acts as a backup to the TVOR--also recently evolved?--to deal primarily with translational disturbances of gaze. Frontal-eyed animals with good binocular vision must be able to keep both eyes directed at the object of regard irrespective of proximity and, in order to achieve this during translational disturbances, the output of the TVOR is modulated inversely with the viewing distance. This sensitivity to absolute depth is also shared by OKNe, consistent with the idea that OKNe is synergistic with the TVOR and shares some of its central pathways. There is evidence that OKNe is also sensitive to relative depth cues such as motion parallax and disparity, which I suggest help the system to segregate the object of regard from other elements in the scene. I also suggest that highly complex optic flow patterns (such as those experienced by the moving observer who looks a little off to one side of his direction of heading) are dealt with by a third visual tracking mechanism--the smooth pursuit system--that spatially filters visual motion inputs so as to exclude all but the motion of the object of interest (local optic flow). PMID- 8420561 TI - The role of visual motion in the stabilization of body posture. PMID- 8420562 TI - Movement detection in arthropods. PMID- 8420563 TI - Directional selectivity in vertebrate retinal ganglion cells. PMID- 8420564 TI - New strategies for winning more battles. PMID- 8420565 TI - Demographics map route to MSV membership growth. AB - The Medical Society of Virginia's new database revealed 1) an aging membership with a high percentage of dues exemptions; 2) an apparent reluctance on the part of many young physicians to consider MSV dues a worthwhile investment; and 3) a marked underrepresentation in the MSV membership of women, who now comprise nearly half of most medical school classes. In order to maintain a viable organization, the Medical Society of Virginia must do more to attract those who are underrepresented in the membership. This is not a choice. It is an imperative. PMID- 8420566 TI - Expect calls from pharmacists as DUR process gears up. PMID- 8420567 TI - Resection for esophageal carcinoma at a Virginia hospital 1981-1989. AB - Between January 1, 1981 and December 31, 1989, 222 patients with carcinoma of the esophagus were seen at Fairfax Hospital. Fifty-eight (26.1%) underwent esophagogastrectomy. Operative (30-day) mortality was 8.6%. Follow-up was 98.3% complete. Of hospital survivors, 38 (76%) were resected for potential cure versus 12 (24%) for palliation. Consistent with the experience of others, a minority of patients (26%) presented with early (Stage I & II) disease; forty patients (69%) were noted to be Stage III or IV at time of resection and three patients (5%) were stage indeterminant. The five year Kaplan-Meier product limit survival estimate for Stage II patients was 52%, versus 22% for stage III, and 0% for Stage IV. PMID- 8420568 TI - Post-concussive disorders. PMID- 8420569 TI - Fractures in elderly psychiatric patients: 42-month study. PMID- 8420570 TI - Urges adoption by all states of Virginia's SB 499. PMID- 8420571 TI - Structure and function of eukaryotic proprotein processing enzymes of the subtilisin family of serine proteases. AB - Production of a broad spectrum of regulatory proteins in eukaryotes occurs via an intricate cascade of biosynthetic and secretory processes. Often these proteins initially are synthesized as parts of higher molecular weight, but inactive, precursor proteins. Specific endoproteolytic processing of these proproteins is required to generate the regulatory proteins in a mature and biologically active form. Such endoproteolysis generally occurs at cleavage sites consisting of particular sequence motifs of basic amino acids, often paired basic residues. This phenomenon, first observed almost 25 years ago, has intrigued scientists ever since then. Nevertheless, the responsible enzymes remained elusive for long. The first known eukaryotic enzyme with the exquisite cleavage specificity for paired basic amino acid residues was the prohormone processing enzyme kexin (EC 3.4.21.61), a subtilisin-like serine protease that is encoded by the KEX2 gene of yeast Saccharomyces cerevisiae. Recently, a number of kexin-like mammalian proprotein-processing enzymes were discovered. The enzyme furin, which is encoded by the fur gene, was the first and can be considered the prototype of a mammalian subclass of subtilisin-like serine proteases. It is predicted to contain a "prepro" domain, a subtilisin-like catalytic domain, a middle domain, a cysteine rich region, a transmembrane anchor, and a cytoplasmic domain. Furin is expressed in a wide variety of tissues, perhaps even in all tissues. In all likelihood, it is the enzyme responsible for the proteolytic bioactivation of a wide variety of precursor proteins. Two other novel mammalian proprotein-processing enzymes are PC1 (also known as PC3) and PC2. Some of the protein domains of these enzymes resemble those in kexin and furin, however, there are also differences. The PC1/PC3 and PC2 enzymes exhibit a more restricted expression pattern than furin. It has been suggested that PC1/PC3 and PC2 are involved primarily in the processing of prohormones within the regulated secretory pathway of cells of endocrine and neural tissue. Recently, the coding sequences for two other candidate mammalian proprotein-processing enzymes were identified. They were called PACE4 and PC4. Like that of furin, the tissue distribution of PACE4 is widespread. PC4, however, may represent a candidate for a precursor-processing endoprotease that is specifically expressed in the testicular germ cells. Finally, DNA sequences encoding kexin- and furin-like candidate pro-protein processing enzymes have been identified in Drosophila melanogaster, Dfur1 and Dfur2 genes; in Xenopus laevis, Xen-14 gene; and in Caenorhabditis elegans, bli-4 gene.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8420572 TI - Molecular mechanisms of hematologic malignancies. AB - This article reviews both the role of signal transduction in human cancer and the recently uncovered molecular mechanisms involved in a large number of human hematologic malignancies. Chronic myelogenous leukemia, follicular lymphoma, and colorectal carcinoma are used as models to illustrate the different, as well as partly convergent, molecular pathways involved in the multistep process of human neoplasias. Examples are also provided as to how unique molecular markers can be used as fast, sensitive, and precise tools for cancer prevention, early diagnosis, and the management of residual disease. PMID- 8420573 TI - Ras-responsive genes and tumor metastasis. AB - Transfected ras oncogenes have been shown to induce metastatic properties in some cells. Clarification of the mechanisms by which ras is able to increase the metastatic ability in model systems will improve our understanding of tumor progression to metastasis, even in those cells in which ras activation has not been implicated. Many of the consequences of ras expression also have been detected in cells that have become metastatic in the apparent absence of an altered ras gene, suggesting that there is a set of common changes that can lead to metastasis with multiple signals capable of eliciting these changes. These changes, which have been documented for some ras-transformed cells, include increased expression or activity of various degradative enzymes, including metalloproteinases (type IV collagenases) and cysteine proteinases (cathepsins L and B), as well as decreased expression or activity of their inhibitors (TIMPs and cystatins, respectively). In addition, some metastatic ras-transformed cells have an increased expression of calcyclin, a cytoplasmic calcium-binding protein, and osteopontin, a secreted calcium-binding protein with possible adhesive function. Not all cells, however, respond in the same fashion to a ras oncogene signal. Some cells are resistant to ras-mediated tumor progression to metastasis. Understanding the mechanism by which these cells fail to respond to a specific oncogene signal may provide clues with broader applicability and potential therapeutic relevance. In this review, we summarize some of the studies in which ras has been used as a tool to learn about the molecular requirements for metastasis. We discuss ras-mediated changes in gene expression and how these may contribute to metastatic ability, as well as some possible mechanisms by which ras expression may result in altered expression of other genes. We also consider some cell lines which appear to be resistant to an oncogenic ras signal and possible mechanisms for this nonresponsiveness. These studies are providing insights into the molecular mechanisms of tumor metastasis and the responses of cells to oncogenic signals. PMID- 8420574 TI - A woman's work... PMID- 8420575 TI - Identification of soft tissue sarcoma deaths in cohorts exposed to dioxin and to chlorinated naphthalenes. AB - Identification of soft tissue sarcomas (STSs) in epidemiologic mortality studies is complicated by nosologic coding rules that require that STSs arising in a visceral organ must be coded in the International Classification of Diseases (ICD) category for that organ, rather than in the ICD category for malignant neoplasms of connective tissue. Moreover, prior studies have shown poor agreement between diagnoses recorded on death certificates compared with those in hospital records for these tumors. We reviewed deaths from STS among workers in a registry of 6,716 dioxin-exposed workers at the National Institute for Occupational Safety and Health (NIOSH) and in a NIOSH cohort mortality study of 10,240 workers exposed to chlorinated naphthalenes. We identified 19 subjects with STSs. Of these, 17 (89%) were identifiable by reading the entries on selected death certificates, and two (11%) were found only by reviewing medical records of cases coded to ICD categories likely to have contained STS. Of the 17 STSs identified from death certificates, only nine (53%) had been coded as underlying cause of death to the ICD category "malignant neoplasms of soft and connective tissue." Medical records were obtained for 14 of the 17 cases (82%), and in each case, the STS diagnosis was verified. Tissue blocks from tumors were available for review in nine of the 17 cases identified from death certificates, and the diagnosis of STS was verified in seven (78%). Nosologic rules reduce the sensitivity of cohort mortality studies to detect excesses of STS.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420577 TI - Misclassified sarcomas and confounded dioxin exposure. PMID- 8420576 TI - Estrogen receptor status and dietary intakes in breast cancer patients. AB - We used data from a study of racial differences in cancer patient survival to examine the association between estrogen receptor status and the intake of nutrients and food groups among 689 black and white women, ages 20-79, with breast cancer newly diagnosed in 1985 and 1986. We reviewed medical records and collected interview data, including a 34-item food frequency questionnaire. Consistent with published reports, we found positive estrogen receptor status to be positively associated with age and inversely associated with parity and oral contraceptive use. Whites were more likely than blacks to have estrogen receptor positive tumors. We examined eight nutrients and six food groups in multivariate analyses for association with estrogen receptor status. After adjusting for age, race, usual body mass index, and parity, a high percentage of calories from fat was associated with estrogen receptor-positive cancer, and a high percentage of calories from carbohydrates was associated with estrogen receptor-negative breast cancer. These findings indicate that women with breast cancer who are on diets with a high percentage of calories from fat may be more likely to develop estrogen receptor-positive cancers. PMID- 8420578 TI - Additive risk versus additive relative risk models. AB - The distinction between additive risk models and additive relative risk models is important when nonadditivity is used as a criterion for interdependence of causal effects (causal interaction). I show here that, in stratified studies, additive relative risk models do not provide the often-assumed correspondence between additivity and absence of causal interaction. Under the causal models of Rothman and others, complete assessment of causal interaction requires that one fit additive risk models; in matched case-control studies, such fitting may require external information. PMID- 8420579 TI - Measures of effect based on the sufficient causes model. 1. Risks and rates of disease associated with a single causative agent. AB - The sufficient causes model of disease occurrence leads to a specific conception of how the risk of disease resulting from exposure to an agent combines with background risks. From this conception, one can derive a method for quantifying in populations the respective effects of the agent and background, using either rates or risks. This method differs from the usual difference or ratio measures of effect by taking into account the probability that the sufficient cause of disease involving the agent of interest and that not involving it will both occur during the observation period. The method leads to: (1) measures of the risks or rates of completion of sufficient causes involving or not involving the agent of interest; (2) a measure of the proportion of cases preventable by removing or blocking the agent, based on observed risks of disease. This proportion varies as a function of the duration of exposure; (3) a measure of the proportion of cases caused by the agent, based on observed rates of disease. This proportion is constant over time, if the rates are; (4) a causal interpretation of a constant rate ratio, when the rates vary over time. PMID- 8420580 TI - Surveillance bias and the excess risk of malignant melanoma among employees of the Lawrence Livermore National Laboratory. AB - To assess the role of surveillance bias in the observed three-fold excess of cutaneous malignant melanoma (CMM) at the Lawrence Livermore National Laboratory (LLNL) in California, we examined the thickness of CMMs among all 20 laboratory employees who were members of a large prepaid health plan and whose CMM was diagnosed from 1970 through 1984. For comparison, we reviewed slides of 36 other members of the same health plan matched (usually 2:1) to the laboratory case by age, sex, facility, and year of diagnosis. Three expert dermatopathologists read the slides using a multiheaded microscope to reach a consensus; they were blind to the laboratory employment status of the subjects. We found that from 1970 to 1976, before there was widespread publicity about the excess incidence of CMM at LLNL, lesion thickness was greater for non-LLNL employees (mean difference = 1.5 mm; 95% confidence interval 0.1-2.9). From 1977 through 1984, however, there was no appreciable difference [mean difference = -0.3 mm; 95% confidence limits (CL) = -1.4, 0.9]. Dropping the matching to adjust for histologic type of melanoma as well as gender, year, and age at diagnosis yielded substantially the same results. These data are compatible with an effect of surveillance bias up to around 1976, but in this health plan population, they do not support a role for surveillance bias in the continuing excess incidence observed since that time. PMID- 8420581 TI - The effect of parity on the relation between maternal history of spontaneous pregnancy loss and the risk of sudden infant death syndrome in offspring. AB - We conducted a population-based case-control study, using the 1984-1989 linked Washington State birth and death certificate data, to determine whether maternal prior spontaneous pregnancy loss was associated with the risk of sudden infant death syndrome (SIDS). We identified 872 autopsied SIDS cases and 3,704 controls matched to cases on year of birth. To reduce the confounding from unmeasured socioeconomic and behavioral factors, we analyzed the data separately for infants with and without father's race provided on the birth certificate. Among infants whose father's race was known, those of multiparous women with a prior spontaneous pregnancy loss before 20 weeks gestational age were at reduced risk of SIDS, after adjustment for maternal age, prenatal smoking,a nd gravidity. An increased number of prior pregnancy losses was associated with a further reduction in the risk of SIDS: one prior pregnancy loss was associated with a 25% decrease [odds ratio (OR) = 0.75; 95% confidence interval (CI) = 0.52-1.08], and two or more prior pregnancy losses was associated with a 47% decrease (OR = 0.53; 95% CI = 0.30-0.94). There was no such association, however, among infants of women with no prior livebirth. Similar associations were observed between prior spontaneous pregnancy loss during the entire gestational period and the risk of SIDS in offspring. To explain the different associations between maternal prior pregnancy loss and the risk of SIDS observed among infants of women with and without a prior livebirth, we hypothesize an upward bias resulting from the presence of a larger proportion of infants of human leukocyte antigen-compatible couples among first liveborn infants. PMID- 8420582 TI - Patterns of pesticide use among farmers: implications for epidemiologic research. AB - Epidemiologic studies of farmers have linked pesticides with certain cancers. Information on exposures from many of these studies was obtained by interview of farmers or their next-of-kin. The reliability and validity of data on pesticide use obtained by recall, often years after the event, have been questioned. Pesticide use, however, is an integral component in most agricultural operations, and the farmers' knowledge and recall of chemicals used may be better than for many other occupations. Contrary to general belief, many farmers typically use only a few pesticides during their lifetimes and make only a few applications per year. Data from U.S. Department of Agriculture surveys indicate that herbicides are applied to wheat, corn, soybeans, and cotton and that application of insecticides to corn averages two or fewer times per year. In epidemiologic studies at the National Cancer Institute, the proportion of farmers ever reporting lifetime use of five or more different chemicals was 7% for insecticides and 20% for herbicides. Surrogate respondents have often been used in epidemiologic studies of cancer; they are able to recall pesticide use with less detail than the farmers themselves. The pesticides reported by surrogates were the same as reported by subjects themselves, but with less frequency. Comparison of reporting by cases and controls provided no evidence of case response (differential) bias; thus, inaccurate recall of pesticide use by subjects or surrogates would tend to diminish risk estimates and dilute exposure response gradients. PMID- 8420583 TI - AIDS in sub-Saharan Africa: the epidemiology of heterosexual transmission and the prospects for prevention. AB - As the epidemic of the acquired immunodeficiency syndrome (AIDS) in sub-Saharan Africa enters its second decade, much has been learned about the distribution and determinants of the disease and its causative agent, the human immunodeficiency virus (HIV). Over 6 million people, or 2.5% of the adult population, are thought to be infected with HIV. The distribution of HIV is largely determined by sexual behavior; as for other sexually transmitted diseases, the characteristics of sexual networks determine the extent and rate of spread of HIV. Female sex workers and their male clients are at high risk for HIV and have been important in initiating the epidemic in many African countries. The dynamics of HIV in the rest of the population are complex; men with multiple sexual partners are largely responsible for transmission of HIV to women in the general population. Other sexually transmitted diseases and lack of male circumcision may increase the probability of transmission of HIV during sexual intercourse and probably are partially responsible for the rapid diffusion of HIV in Africa. Interventions among high-risk groups are needed, but they must be accompanied by attempts to induce behavior change among men and women in the general population. Epidemiologic studies of the determinants of sexual behavior and sexual contact patterns, as well the design and evaluation of interventions, are urgently needed. Key areas for development are the study of behavioral exposures and outcomes, the evaluation of interventions, developing new methods for conducting interventions in resource-poor environments, and increasing the number of African scientists with the skills and resources to conduct epidemiologic studies. PMID- 8420585 TI - Pregnancy loss and work schedule during pregnancy. AB - Shift work has been associated with many health complaints, but its relation with pregnancy loss has rarely been studied. We compared the work schedule during pregnancy of 331 women with a pregnancy loss with that of 993 pregnant women matched on gestational age. The risk of pregnancy loss was four times higher among women with fixed evening work schedules in comparison with women on a fixed day schedule (odds ratio = 4.17; 95% confidence interval = 2.19-7.92), and more than twice as high among those on a fixed night schedule (odds ratio = 2.68; 95% confidence interval = 0.53-13.43). PMID- 8420584 TI - The mortality experience of workers exposed to 2,3,7,8-tetrachlorodibenzo-p dioxin in a trichlorophenol process accident. AB - Epidemiologic studies on dioxin, specifically 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), have not consistently found an increased cancer risk. This study examines mortality rates for 754 employees at a chemical plant after an accident in 1949 in which 122 workers developed chloracne from high exposure to TCDD. We also examined exposure to 4-aminobiphenyl, a potential bladder carcinogen. Although the numbers are small, and confounding factors cannot be ruled out, workers exposed to 4-aminobiphenyl who developed chloracne from the accident had increased mortality rates from soft tissue sarcoma, bladder cancer, and respiratory cancer. All soft tissue sarcomas occurred among workers with potential 4-aminobiphenyl exposure, whereas no soft tissue sarcomas occurred among workers with TCDD exposure alone. In these workers, exposure to TCDD alone at levels great enough to cause chloracne was not associated with increased cancer rates. PMID- 8420586 TI - Epidemic asthma in Cartagena, Spain, and its association with soybean sensitivity. AB - We conducted a matched case-control study to assess the association between epidemic asthma and soybean sensitivity in Cartagena, Spain. In skin tests, 81% of the cases and no control subjects were positive to the commercial soybean allergen; 87% of the cases and 12% of the controls were positive to the prepared soybean allergen; and 69% of the cases and 6% of the controls showed high levels of specific immunoglobulin E antibodies (odds ratio = 10; 95% confidence interval = 1.4-433). The association remained strong after adjustment for total immunoglobulin E and for 28 other allergens. The results indicate that soybean sensitivity may be causing asthma epidemics in Cartagena. PMID- 8420587 TI - Sex differences in coronary heart disease mortality trends: the Minnesota Heart Survey, 1970-1988. AB - Recent U.S. national statistics indicate a slowing of the rate of decline in coronary heart disease (CHD) deaths among women. To examine recent sex-specific rates of mortality decline in the Minnesota Heart Survey, we computed the average annual percentage change in age-adjusted CHD death rate for the periods 1970-1978 and 1979-1988. We fit a log-linear regression model to the yearly CHD death rates, with separate sex-specific slopes estimated simultaneously for the two calendar periods. The average annual percentage decline in age-adjusted CHD death rate among men was slightly greater in the period 1979-1988 [4.8%; 95% confidence interval (CI) = 5.2-4.3] than in the period 1970-1978 (3.9%; 95% CI = 4.6-3.1). In contrast, among women, the rate of decline was less in the period 1979-1988 (3.6%; 95% CI = 4.8-2.3) than in 1970-1978 (4.8%; 95% CI = 5.6-4.0). We found a slowing of the rate of decline in out-of-hospital CHD deaths in both men and women. We observed a large increase in the rate of decline in the CHD death rate in hospital among men in the period 1979-1988 (8.3%; 95% CI = 9.3-7.2), compared with the period 1970-1978 (2.4%; 95% CI = 3.2-1.7). This increase did not occur among women (4.1%; 95% CI = 5.7-2.5 in 1970-1980 vs 3.8%; 95% CI = 4.9-2.7 in 1970-1978); this difference is a major factor in the steeper recent decline in overall CHD mortality among men compared with women. PMID- 8420588 TI - Community-based surveillance of acute ischemic heart disease: are one-time mailed questionnaires to physicians useful? AB - A 12-month data collection from medical records in Seattle/King County for acute ischemic heart disease among patients with diabetes mellitus was augmented with mailed questionnaires to personal physicians. The responsible Human Subjects Committee restricted the study to a one-time questionnaire mailing. Only 380 of 1,235 patients required physician contact when information was missing from primary data sources. Questionnaires were highly personalized, and most were limited to one page. Seventy-seven per cent of 330 physicians returned at least one questionnaire, and 62% of all questionnaires were returned with usable data. We conclude that useful data may be collected from physicians by way of mailed questionnaires if researchers use a brief and personal format. PMID- 8420589 TI - Subjective terminology. PMID- 8420590 TI - Subjective terminology. PMID- 8420591 TI - Subjective terminology. PMID- 8420592 TI - The delusional misidentification syndromes in patients with and without evidence of organic cerebral disorder: a structured review of case reports. AB - Two series, each of 50 cases of delusional misidentification reported in the literature, were analyzed in order to study the interaction between organic and functional mental processes. The details of the time course of the development of the delusional misidentification and related mental symptoms, the phenomenology, and evidence of cognitive impairment and/or cerebral damage were recorded. The first series of cases included delusional misidentification either of place, and/or of persons; the second series was limited to cases, published since 1977, with delusional misidentification of person, who had had an electroencephalogram (EEG) and/or computerised tomographic (CT) brain scan. Paranoid delusions, preceding the onset of the delusional misidentifications, were more common in cases without evidence of organic cerebral disorder. In the second series there was good evidence of an inverse relationship between the presence of paranoid delusions preceding the delusional misidentification, and the intensity of organic cerebral disorder; (1) no evidence clinically or on investigation, (2) evident only on investigation, or (3) evident clinically. In the first series delusional misidentification of place was more common in cases with evidence of organic brain disorder, whereas delusional misidentification of person was more common with functional mental disorder. Nevertheless the delusional misidentification was equally likely to involve objects of personal significance whether or not there was evidence of organic cerebral disorder. Paramnesic misidentifications were associated with both memory impairment and disorientation for time. PMID- 8420593 TI - A comparison of the behavioral effects of oral versus intravenous mCPP administration in OCD patients and the effect of metergoline prior to i.v. mCPP. AB - In prior studies form three centers, an exacerbation of obsessive-compulsive disorder (OCD) symptoms was reported in some (55%-83%) patients with OCD receiving the serotonergic agonist m-chlorophenylpiperazine (m-CPP) orally, whereas intravenously administered mCPP produced anxiety but no OCD symptom exacerbation. In the present replication attempt, 27 OCD patients were given mCPP either orally (n = 17) or intravenously (n = 10) under double-blind conditions, using identical behavioral rating measures. OCD symptoms were significantly increased after intravenous mCPP (0.1 mg/kg), but not after oral mCPP (0.5 mg/kg). Anxiety and other ratings were markedly elevated after intravenous mCPP administration. After oral mCPP administration, anxiety and most other self ratings were only slightly elevated in comparison to placebo administration, and behavioral rating increases were no different for the OCD patients compared to age-matched healthy controls. Pretreatment with the potent serotonin (5-HT) antagonist, metergoline, prior to intravenous mCPP was associated with essentially complete blockade of the exacerbation in OCD symptoms and the other behavioral responses in the OCD patients. These results suggest that the behavioral response of OCD patients to mCPP are variable and depend on the route and dose of mCPP. In addition, the ability of metergoline to antagonize the behavioral effects of intravenous mCPP suggests that these responses are mediated by 5-HT1/5-HT2 receptors. PMID- 8420594 TI - The cholinergic REM sleep induction test with pilocarpine in mildly depressed patients and normal controls. AB - Previous studies suggested that depressed patients enter rapid-eye movement (REM) sleep more quickly than normal controls following the administration of muscarinic agonists such as arecoline or RS 86. We recently reported that pilocarpine, an orally active muscarinic agonist, induced REM sleep and reduced Stage 3 & 4 (Delta) sleep in young normal volunteers. In this study we present preliminary evidence that pilocarpine had similar effects on REM latency, REM percentage, and Delta (Stages 3 & 4) sleep percentage in depressed patients and controls. Pilocarpine, however, decreased stage-4 sleep in controls more than in depressed patients. Because this group of patients were only mildly depressed at the time of the study, had a high frequency of comorbid substance abuse diagnoses, and had normal electroencephalogram (EEG) sleep patterns under placebo conditions, further studies are necessary to test the hypothesis that depressed patients show hypersensitive cholinergic REM sleep induction. PMID- 8420595 TI - meta-Chlorophenylpiperazine decreases slow-wave sleep in humans. PMID- 8420596 TI - Folate, B12 and thyroid function in depression. PMID- 8420597 TI - Sleep deprivation with consecutive sleep-phase advance therapy in patients with major depression: a pilot study. PMID- 8420598 TI - Cortisol/18-OH-DOC in menstrual cycle. PMID- 8420599 TI - Cell cycle characteristics and schizophrenia. PMID- 8420600 TI - Cryptosporidiosis. PMID- 8420601 TI - Human T-cell responses in Leishmania infections. PMID- 8420602 TI - Evolution of prosobranch snails transmitting Asian Schistosoma; coevolution with Schistosoma: a review. PMID- 8420603 TI - Toxoplasmosis: an update on clinical and therapeutic aspects. PMID- 8420604 TI - Anisakidae and anisakidosis. PMID- 8420605 TI - Predictors of future breast cancer risk. PMID- 8420606 TI - Canadian National Breast Screening Study: first screen results as predictors of future breast cancer risk. AB - Characteristics of women aged 40-59, recorded at an initial breast screen, were compared with the outcome of incident, invasive breast cancer occurring up to 3 years after the initial screen. The study design was case-control, nested within the study population of the Canadian National Breast Screening Study. Screening consisted of a two-view mammographic examination and physical examination of the breasts; additional risk factor information was obtained from self-administered questionnaires. Of the etiological risk factors considered only age at entry or years menstruating were significant risk factors for breast cancer (P < 0.0025). Years menstruating accounted for much of the age effect and all of the menopausal status effect. Risk factors obtainable from mammography and physical examination were more significant than self-reported risk factors, other than age or years menstruating. In spite of possible misclassification of the variable parenchymal pattern, women with a mammographic film classified as either P2 or DY had a 2 fold risk of breast cancer (odds ratio = 2.1; 95% confidence interval = 1.5-2.9, P = 2.9E-05). An abnormality reported by either the radiologist [odds ratio = 1.7 (1.3-2.3)] or nurse examiner [odds ratio = 1.9 (1.4-2.6)] was also associated with an independent significant risk. PMID- 8420607 TI - Serological precursors of cancer: serum hormones and risk of subsequent prostate cancer. AB - A population-based nested case-control study was conducted to determine the relation of prediagnostic serum levels of testosterone, dihydrotestosterone, prolactin, follicle-stimulating hormone, luteinizing hormone, estrone, and estradiol to the risk of subsequent prostate cancer. Serum specimens of study subjects were available from a blood collection campaign in Washington County, Maryland, in 1974. Serum hormone levels of 98 histologically confirmed prostate cancer cases diagnosed in the subsequent 13 years were compared to those of 98 controls who were individually matched to cases on the basis of age (within weeks), sex, and race. There were no significant differences in levels of these hormones between cases and controls, although elevated levels of luteinizing hormone and of testosterone:dihydrotestosterone ratios were associated with mild increased risks of prostate cancer. PMID- 8420608 TI - ASPO Distinguished Achievement Award Lecture. Studies on the epidemiology and natural history of benign breast disease and breast cancer using nipple aspirate fluid. AB - Nicholas L. Petrakis, M.D., was presented the American Society of Preventive Oncology's Distinguished Achievement Award at the Society's annual meeting on March 15, 1992, in Bethesda, Maryland. The basis for choosing Dr. Petrakis was his distinguished achievements for over 20 years in research and education concerning the etiology and prevention of cancer, with substantial emphasis on breast cancer. Dr. Petrakis obtained a Bachelor of Arts degree from Augustana College, Sioux Falls, South Dakota, and his Doctor of Medicine degree from Washington University, St. Louis, Missouri. He has held numerous principal positions at the University of California, beginning with a position with the U.S. Public Health Service, National Cancer Institute Laboratory of Experimental Oncology, and then the Cancer Research Institute, both at the University of California, San Francisco. Dr. Petrakis has held continuing appointments in the Department of Preventive Medicine and Department of Epidemiology and International Health, beginning as an Assistant Professor. He was subsequently promoted to Associate Professor and Professor of Preventive Medicine. During the years from 1978 to 1989, Dr. Petrakis was chair of the Department of Epidemiology and International Health. Since 1981 he has been Professor of Epidemiology, University of California, Berkeley, School of Public Health. In 1989 he became Professor Emeritus of Preventive Medicine and Epidemiology.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420609 TI - Serum levels of dehydroepiandrosterone and dehydroepiandrosterone sulfate and the risk of developing gastric cancer. AB - Although the incidence of gastric cancer varies widely between countries it is nonetheless a leading cause of cancer deaths worldwide. Migration studies indicate that dietary choices are an important exogenous factor. The United States has a very low incidence of gastric cancer, suggesting that exogenous etiological agents are at a minimum and providing a favorable setting for detecting important endogenous etiological factors. Dehydroepiandrosterone and dehydroepiandrosterone sulfate are endogenous steroids produced in the adrenal gland. Epidemiological studies show that the risk of developing specific cancers is related to the serum or urinary levels of these steroids. In addition, dehydroepiandrosterone prevents a variety of spontaneous and chemically induced tumors when administered to laboratory animals. To examine the association between circulating levels of dehydroepiandrosterone and dehydroepiandrosterone sulfate and the development of gastric cancer, we measured the serum levels of these steroids in 13 individuals who donated serum to the Washington County Maryland serum bank in 1974 and who subsequently developed gastric cancer and in 52 matched controls. Prediagnostic serum levels of dehydroepiandrosterone were 38% lower in cases as compared to controls (P = 0.09). The risk of developing gastric cancer increased with decreasing levels of both steroids. Adjustment for confounding factors such as smoking or the interval between blood donation and time to diagnosis did not alter the findings. These results suggest that there may be a role for this steroid in the prevention of gastric cancer. PMID- 8420610 TI - Helicobacter pylori antibodies in areas of Italy at varying gastric cancer risk. AB - In a survey of 930 adults aged 35-74 years randomly sampled from the general population of four areas of Italy, two at low and two at high risk for gastric cancer, plasma levels of Helicobacter pylori IgG antibodies were assayed in order to investigate associations with the geographical distribution of gastric cancer and other dietary and life-style factors, as assessed by personal interview. H. pylori positivity (antibody titer above or equal to 10 micrograms/ml), 45% overall, increased with age and was inversely associated with social class but showed little geographical variation or association with dietary variables and blood nutrients. H. pylori positivity was also associated with increased blood levels of pepsinogens, particularly pepsinogen II. The authors discuss these findings in relation to those from a previous case-control study of gastric cancer in the same areas. PMID- 8420611 TI - Plasma selenium concentration predicts the prevalence of colorectal adenomatous polyps. AB - The objective of this cross-sectional study was to determine whether plasma selenium concentration predicts the prevalence of adenomatous polyps of the colon and rectum. The source population for the study was 101 patients undergoing sequential colonoscopies at the Veterans Affairs Medical Center, Tucson, AZ. The study population was then limited to the 48 patients (all male) undergoing their initial colonoscopy who did not have a diagnosis of colorectal cancer. For each of these patients, a prediagnostic fasting plasma selenium concentration was determined. The data from this study suggest that fasting plasma selenium concentrations may be an important risk factor for colorectal adenomas. Patients with fasting plasma selenium concentrations below the median (< 128 mcg/liter) were significantly more likely to have one or more adenomatous polyps (prevalence odds ratio 4.2) and more adenomatous polyps (3.5 times) per patient. There was also a suggestion of a more proximal distribution of adenomatous polyps in the patients with a lower level of selenium. These associations were not confounded by age or smoking. The results of this study are consistent with the experimental animal studies, geographic mortality studies, and prospective cohort studies of selenium and colorectal cancer. PMID- 8420612 TI - Urinary trans,trans-muconic acid as an indicator of exposure to benzene in cigarette smokers. AB - Epidemiological studies have shown an association between cigarette smoking and increased risk of myeloid leukemia in smokers. In evaluating this link it is important to note that cigarette smoke contains benzene, among other carcinogens. Since chronic benzene, among other carcinogens. Since chronic benzene exposure causes acute myeloid leukemia in humans, we aimed to determine the uptake and metabolic activation of benzene from cigarette smoke in smokers by measuring the levels of the urinary benzene metabolite, trans,trans-muconic acid (t,t-MA). The method used involved a clean-up procedure, followed by high-performance liquid chromatography with UV detection. The levels of urinary t,t-MA ranged from 0.02 to 1.3 mg/g creatinine, resulting in a mean of 0.29 +/- 0.04 mg/g creatinine in 42 male smokers, and corresponding values in nonsmokers ranged from "nondetectable" to 0.52 mg/g creatinine with an average of 0.09 +/- 0.02 mg/g creatinine. In the current study, the levels of t,t-MA in smokers were about 3 times higher than those in nonsmokers (P = 0.0001), and a significant correlation between concentration of t,t-MA and levels of cotinine in smokers was observed (r = 0.55; P = 0.0001; 95% confidence interval, 0.30-0.93), suggesting that urinary t,t-MA can be utilized as a biochemical marker to quantitate benzene exposure due to cigarette smoking. PMID- 8420613 TI - Polycyclic aromatic hydrocarbon-DNA adducts in white blood cells and urinary 1 hydroxypyrene in foundry workers. AB - In an ongoing comprehensive evaluation of biological markers, workers in or near an iron foundry with varying exposure to polycyclic aromatic hydrocarbons (PAH) were analyzed for molecular response to this exposure. Exposure to benzo(a)pyrene, determined by personal monitors worn by the workers (2 to 60 ng/m3), was considerably lower than in a previous study at this foundry (< 50 to 200 ng/m3) (F.P. Perera et al., Cancer Res., 48: 2288-2291, 1988). Two biomarkers, 1-hydroxypyrene in urine measured by high-performance liquid chromatography with fluorescence detection (a measure of internal dose) and PAH DNA adducts in WBC measured by immunoassay (a measure of biologically effective dose) were assessed to demonstrate their relationship to the lowest exposures yet analyzed in foundry workers. In addition, these markers were analyzed for dose response and interindividual variability. Cigarette smoking, but not age or charbroiled food, influenced the level of 1-hydroxypyrene but not PAH-DNA adducts. When workers were classified into three exposure categories (low, medium, and high), mean 1-hydroxypyrene levels were 2.7, 1.8, and 3.6 mumol/mol creatinine, respectively. Comparisons by analysis of variance showed a significant difference between the groups after controlling for smoking (P = 0.02), but a trend test using multivariate linear regression analysis was not significant (r = 0.27; P = 0.07). Substantial interindividual variation was demonstrated by the 19- to 20-fold range in the values within each of the three exposure groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420614 TI - 4,4'-Methylene-bis(2-chloroaniline)-DNA adduct analysis in human exfoliated urothelial cells by 32P-postlabeling. AB - 4,4'-Methylene-bis(2-chloroaniline) (MOCA) is an aromatic amine used widely in industry, and human exposure to this compound is well documented. MOCA induces lung and liver tumors in rodents and urinary bladder tumors in dogs, and it is regarded as a suspect urinary bladder carcinogen in humans. In this pilot study, we investigated the occurrence of MOCA-DNA adducts in exfoliated urothelial cells of a MOCA-exposed worker by 32P-postlabeling analysis. Urine samples were collected from the worker at various times after accidental acute exposure to MOCA. DNA isolated from exfoliated urothelial cells collected from each urine sample was enzymatically digested and postlabeled with excess [32P]ATP. Thin layer chromatographic analysis of the labeled digests revealed the presence of a single, major DNA adduct that cochromatographed with the major N-hydroxy-MOCA-DNA adduct, N-(deoxyadenosin-8-yl)-4-amino-3-chlorobenzyl alcohol, formed in vitro. The MOCA-DNA adduct was detected in samples obtained between 4 and 98 h after initial exposure but not in samples collected at later times. The level of DNA adducts 4 h after exposure was determined to be 516 adducts/10(8) nucleotides. A 5-fold decrease in adduct level was observed 14 h later, followed by a gradual decrease over subsequent days. The results indicate that MOCA is potentially genotoxic to human urinary bladder in vivo and that 32P-postlabeling analysis of exfoliated urothelial cells provides a noninvasive means for biomonitoring the formation of MOCA-DNA adducts resulting from occupational exposure. PMID- 8420615 TI - A randomized double-blind intervention study on the effect of calcium supplementation on esophageal precancerous lesions in a high-risk population in China. AB - To determine whether dietary calcium supplementation affects esophageal precancerous lesions, 200 subjects with esophageal lesions in a high-risk area for esophageal cancer in China (Huixian, Henan) were randomly divided into 2 groups (100 subjects/group). Subjects in one group received an oral supplementation of calcium carbonate tablets (1200 mg of calcium daily), and subjects in the other group received placebo pills for 11 months. At the entry and the end of the trial, esophagoscopy was performed, and 2 or 3 biopsy specimens were taken from the middle and lower thirds of the esophagus and from macroscopic lesions, if any, of each subject for histopathology and cell proliferation analysis with deoxythymidine labeling. In comparison to normal epithelium, increased proliferative compartment size was observed in epithelia with hyperplasia or dysplasia. After the intervention, the percentage of individuals with "normal epithelium," "basal cell hyperplasia," "basal cell hyperplasia II," and "basal cell hyperplasia III and dysplasia" were 44, 31, 13, and 11% in the calcium group and 35, 39, 17, and 6% in the placebo group, respectively. The labeling index was 0.046 in the calcium group and 0.044 in the placebo group. After the intervention, the labeling index in basal cell layers 1 to 5, the major zone of cell proliferation, fell 38% in the calcium group and 44% in the placebo group from before the intervention. Therefore, in this study, calcium supplementation was not shown to have beneficial effects in alleviating precancerous lesions and abnormal cell proliferation patterns. PMID- 8420616 TI - Epidemiology of ovarian cancer. PMID- 8420617 TI - Differential lateral septal vasopressin innervation in aggressive and nonaggressive male mice. AB - The vasopressinergic (VP) projection from the bed nucleus of the stria terminalis (BNST) to the lateral septum (LS) is sexually dimorphic and dependent of androgens at adult and neonatal age. We studied the relation between testosterone (T) and VP in male mice, which were genetically selected for their differences in aggression level. Aggressive males, characterized by a short attack latency (SAL), have a higher production capacity of T at adult age compared to males with a long attack latency (LAL). Neonatally, however, a higher T production occurs in the nonaggressive LAL males than in SAL males. In the present study we showed a more dense VP-immunoreactive (VP-ir) innervation in the LS and a higher VP-ir neuron density in the BNST of LAL males as compared to SAL males. The described differences may be the consequence of a differential neonatal androgen effect on the organization of the forebrain vasopressinergic network. PMID- 8420618 TI - The effects of stress on splenic immune function are mediated by the splenic nerve. AB - Intermittent footshock (FS) suppresses immune function of spleen cells. To determine if the autonomic nervous system mediates this immunosuppression in spleen cells, we tested whether cutting the splenic nerve, which depletes splenic norepinephrine levels by 98-100% and eliminates catecholamine fibers, blocks the effects of stress. Splenic nerve sections, sham operations, or no surgery were performed on male Sprague-Dawley rats. Ten days later, rats were injected with sheep red blood cells (SRBC). Three days later, rats were placed in a chamber equipped with a shock grid. Foot shock (1.6 mA) was administered for 5 s on a VI 3.5 min schedule for 60 min. Each FS was preceded by a 15-s warning tone. Controls were treated identically except for the FS. The next day spleen cells were harvested and the number of IgM plaque-forming cells (PFCs) determined. For the sham and unoperated control animals, the number of PFCs was reduced for the stressed animals relative to the nonstressed controls, and there was no effect of the sham surgeries. In contrast, there was no difference between the stressed and nonstressed groups in which the splenic nerve had been sectioned, and their PFC response was comparable to the controls. Next we examined the effects of FS on the proliferative response to mitogens (PHA and ConA) following splenic nerve sections or sham operations. One week following surgery, animals were given a 60 min session of FS or exposed to the chamber/tone without FS. Rats were then killed, spleens harvested, and the proliferative response to mitogens determined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420619 TI - Decrease of central dopamine level in the adult spontaneously hypertensive rats related to the calcium metabolism disorder. AB - The metabolism of calcium and brain dopamine in spontaneously hypertensive rats (SHR) after the development of hypertension was investigated as a possible model for the hypertension mechanism. Serum calcium level in SHR was lower than that in the normotensive control. Wistar Kyoto rats (WKY, the parent strain of SHR). Conversely, bone calcification of SHR was higher than that in WKY. Possible mechanisms for the lower serum calcium level seen in SHR include a decrease in the availability of calcium from bone. The immunohistochemical dopamine levels in the neostriatum and nucleus accumbens in SHR were lower than those in WKY. In these regions, the dopamine level was increased by the intraventricular administration of CaCl2 through a central, calmodulin-dependent system. This study suggests, based upon previous pharmacological studies, that the decrease of the serum calcium level in SHR causes a decrease in central, calcium-calmodulin dependent dopamine synthesis and a subsequent low level of dopamine in the brain that produces an increase in blood pressure through functions of cerebral dopaminergic neurons and peripheral sympathetic nerves. Our results suggest that this could be one of the mechanisms of hypertension in SHR. PMID- 8420620 TI - Control of rapid elbow extension movement. AB - Normal human subjects made rapid extensions of the elbow from an initial position to visual targets at different angular distances (9 degrees-54 degrees). We recorded joint angle, acceleration, and surface electromyograms (EMGs) with a monopolar electrode from the triceps and the biceps brachii muscles. Both the agonist triceps and antagonist biceps initiated their activation with a slow EMG wave (AGslw and ANTslw). The amplitude of the AGslw increased with angular distance moved, but its duration remained constant. The amplitude of the ANTslw varied among subjects with distance moved. The duration of the ANTslw was approximately constant but more variable than that of the AGslw. The onset of the ANTslw was delayed with increasing movement distance. Our results indicate that movement distance is controlled, at the motoneuron pool level, by the interval of a set of pulses sent to the agonist and antagonist muscles in association with pulse amplitude modulation to the agonist muscle. PMID- 8420621 TI - Area postrema: gastric vagal input from proximal stomach and interactions with nucleus tractus solitarius in cat. AB - The responses of neurons in the area postrema (AP) during electrical stimulation of the gastric vagal branches that serve the proximal stomach were evaluated in halothane-anesthetized cats. The evoked orthodromic responses were localized bilaterally in the AP and had a mean latency of 272 ms (SD +/- 53.1). Convergence of gastric vagal input on single AP units from afferents in the ventral and dorsal vagal trunks serving the proximal stomach was observed on 13% of gastric vagally evoked neurons. Reciprocal connections between neuronal populations in the nucleus tractus solitarius and the AP were identified electrophysiologically by direct microstimulation of the former structure. Spontaneously AP discharging units showed an increase in frequency after NTS microstimulation. Our study provided evidence that the AP neurons received vagal input from the proximal stomach and suggested a potential role of the AP in the regulation of food intake. PMID- 8420622 TI - Prolonged depression of force developed by single motor units after their intermittent activation in adult cats. AB - The fatigue of fast-twitch, glycolytic mammalian motor units [i.e., type FF; nomenclature of (3)] is dependent, in part, on the stimulation regimen (total number of stimuli, frequency, duty cycle, temporal patterning of stimuli, etc.) used to induce fatigue. To study the effect of the temporal pattern of the stimulus train on the rate and extend of fatigue in single FF units, one theoretically acceptable approach would be to use each motor unit as its own control: i.e., a sequential testing with two fatigue tests that differ only in the temporal organization of their stimuli. The purpose of this communication is to provide evidence that such an approach is not feasible when studying FF units, due to the delayed recovery of force following their repetitive activation. It was shown that 1/s activation of single FF units for only 15 or 45 s with intermittent 40-Hz, 300-ms duration trains significantly reduced their force response to a double-pulse shock for several hours. This finding suggests that in studies designed to test for the effects of different stimulation patterns on the fatigue of single motor units, deeply anaesthetized, reduced animal preparations are not appropriate models for the sequential application of different stimulation regimens to fast-twitch, glycolytic, mammalian motor units. PMID- 8420623 TI - Dissociation of delta EEG amplitude and incidence in rat NREM sleep. AB - The delta (1-4 Hz) EEG of nonREM (NREM) sleep was subjected to period/amplitude analysis in 10 Sprague-Dawley rats. During NREM sleep in the 12-h light period, average delta wave amplitude and delta wave incidence (halfwaves/min) both declined; the curves were biphasic with a plateau across hours 4-6. In contrast, the behavior of amplitude and incidence was strikingly different in dark period NREM sleep. At dark onset, amplitude increased sharply and remained at this elevated level without any significant trend across the 12 hours. Delta incidence was low at dark onset and increased with a strong linear trend. These data point to several experiments to test the mechanisms mediating the behavior of delta wave amplitude at the light-dark transition; they also bear on the homeostatic model of delta sleep. PMID- 8420624 TI - Age-dependent change in the inhibitory effect of an adenosine agonist on hippocampal acetylcholine release in rats. AB - To investigate the possibility that age-dependent deficits in acetylcholine (ACh) release are precipitated by the alteration of endogenous purinergic activities, the effects of (-)N6-phenylisopropyladenosine (PIA), an adenosine agonist, in modulating K+ (25 mM)-induced [3H]ACh release from the hippocampal slices of young (3-6 months old) and old rats (26-30 months old) were examined. In young rats, PIA (0.1-10 microM) caused a dose-related inhibition of [3H]ACh release from the hippocampal slices and a significant reduction in [3H]ACh release was observed in the presence of 1 microM PIA. In old rats, a similar pattern of PIA suppression of K(+)-induced [3H]ACh release was observed; however, a 10-fold higher concentration of PIA (10 microM) was required to elicit a significant inhibition. This age-dependent reduction in responsiveness to PIA may be due to an enhanced endogenous adenosine activity in aged rats leading to downregulation of the adenosine receptors. This notion is supported by the finding that both the adenosine concentration and activity of 5'-nucleotidase, an enzyme partially governing adenosine synthesis, were increased in the hippocampus of old rats as compared to their younger counterparts. PMID- 8420625 TI - Pineal sensitivity to pulsed static magnetic fields changes during the photoperiod. AB - The effect of pulsed static magnetic fields on the rat pineal melatonin synthesis was studied at different times of the photoperiod. Exposure to magnetic fields during mid- or late dark phase significantly suppressed pineal N acetyltransferase activity, the rate-limiting enzyme in melatonin synthesis, as well as the melatonin content in the pineal gland. These parameters were not influenced by magnetic fields when the exposure occurred early in the dark phase or during the day. These results suggest that the responsiveness of the pineal gland to magnetic field perturbations changes throughout the photoperiod. PMID- 8420626 TI - Effect of light intensity on diurnal sleep-wake distribution in young and old rats. AB - During the aging process, the amplitude of the circadian rhythms of many physiological variables is reduced. It has been hypothesized that increasing light intensity during the light phase of the light-dark cycle might result in a reduction of age-related changes in the circadian rhythms. Indeed, in the present sleep-wake study in young and old rats it was found that (a) various parameters, such as the light-dark differences and total amounts of each behavioral state responded positively to changes in environmental light intensity (i.e., age related trends were reversed), (b) in both age groups, the logarithm of light intensity appeared to have a linear dose-response relationship with light-dark differences of the sleep-wake states, (c) the light-dark difference of active wakefulness and quiet sleep of old rats under high light intensity were comparable to those of young rats under low light intensity. The results of the present study suggest that, under appropriate conditions, light could be of clinical use in reducing age-related circadian sleep disturbances in humans. This may, in turn, reduce the use of sedatives in elderly people. PMID- 8420627 TI - Cocaine-induced expression of striatal c-fos in the rat is inhibited by NMDA receptor antagonists. AB - To assess the possible involvement of NMDA receptors in mediating the expression of striatal c-fos by cocaine injection, we investigated the effects of the noncompetitive NMDA receptor antagonists, ketamine and (+)-5-methyl-10,11-dihydro 5H-dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801), as well as the competitive NMDA receptor antagonist, 3-(2-carboxypiperazin-4-yl)-propyl-1 phosphonic acid (CPP), in the perikarya of cocaine-treated rat brains. As previously shown by our group, administration of 20 mg/kg cocaine (IP) resulted in the immunocytochemical expression of the protooncogene in numerous cells of the caudate putamen (dorsal/sensorimotor striatum). A ketamine mixture anesthetic (2 mg/kg), however, administered 30 min prior to cocaine exposure completely blocked such genomic expression. Pretreatment with MK-801 (1 mg/kg) or CPP (5 mg/kg) also interfered, albeit to a lesser extent, with the expression of c-fos by cocaine in awake animals. These results indicate that cocaine induction of cellular c-fos in the caudate putamen is mediated at least in part by NMDA sensitive receptors. PMID- 8420628 TI - Bombesin and neurotensin excite neurons in hypothalamic arcuate nucleus in brain slices: an extracellular single-unit study. AB - Using extracellular single-unit recording in brain tissue slices, the effects of bombesin (Bomb), neurotensin (NT), and their analogs on hypothalamic arcuate (ARC) neurons were tested in this study. Adult ovariectomized Sprague-Dawley rats were used for obtaining the brain slices. Both Bomb and NT in pmol ranges stimulated nearly 70% of ARC neurons tested and the effects were all very significant. [D-Trp11]-NT, a NT analog, behaved more like an analog of NT instead of an antagonist, although [Leu13-psi(CH2NH)-Leu14]-Bomb, a Bomb receptor antagonist, could not block most of Bomb's effects on ARC neurons. The significant effects of Bomb and NT shown in this report indicate that both peptides may play significant roles in the neuroendocrine control system. PMID- 8420629 TI - An electromyographic volley at initiation of rapid isometric contractions of the elbow. AB - When normal subjects made rapid isometric extensions of the elbow to target torque, the agonist electromyograms (EMGs), recorded by a monopolar surface electrode, initiated from an EMG volley which was characterized by a negative potential immediately followed by a positive one. This study examined properties of the EMG volley and determined whether or not it is a movement artifact. Surface EMGs recorded by multiple monopolar electrodes showed that the negative peak of the EMG volley was earliest and greatest in amplitude at the motor point region. The integrated negative potential correlated with peak torque. EMGs recorded by needle and bipolar surface electrodes showed a duration of comparative activity with the EMG volley. To eliminate movements between the skin and monopolar electrode, the skin around the electrode was clamped to a fixture. This left the EMG volley unchanged. The antagonist showed a similar EMG volley, but was less stable in duration and amplitude. Our results demonstrated that the EMG volley is not a movement artifact. PMID- 8420630 TI - Human performance with a seventeen-arm radial maze analog. AB - Results using radial mazes suggest rats may have a greater short-term memory (STM) capacity than the human "magical number seven." We examined spatial STM in humans using a radial maze analog drawn on paper. Subjects were instructed to lift, in "random" order, each of 17 cardboard flaps arranged radially around a center point. Fifteen undergraduate subjects, tested seven trials a day on 2 consecutive days, averaged 15.4 correct choices per trial. Thus, humans perform equally well on a 17-arm radial maze analog as rats do on a 17-arm radial maze, suggesting comparable spatial STM capacities, and perhaps homologous brain substrates for these tasks, in these two species. PMID- 8420631 TI - Opposite effects of intranigral ibotenic acid and 6-hydroxydopamine on motor behavior and on striatal neuropeptide Y neurons. AB - Unilateral lesions of the basal ganglia circuit induce a disequilibrium of motor processing, most obviously expressed by the resulting circling behavior. Compensatory events, which reduce the motor asymmetry, could be accompanied by changes in neurotransmitter/modulator parameters in the involved brain regions. In the present investigation, the effects of an interruption of the striato-nigro thalamic loop by ibotenic acid (IBO)-induced lesions of total substantia nigra (SN) on circling behavior and on striatal neuropeptide Y (NPY) neurons were compared with those after the selective destruction of the dopaminergic nigrostriatal projection with 6-hydroxydopamine (6-OHDA). Directly after the operation, IBO-lesioned rats showed a high circling rate to the side contralateral to the lesion, whereas 6-OHDA-lesioned rats showed ipsiversive circling. With the lesion-induced development of dopamine receptor supersensitivity, 6-OHDA-treated rats, when stimulated with the dopaminergic agonist apomorphine, change their circling direction to the contralateral side. Complete IBO lesions of the SN abolished this effect: rats continued to circle to the contralateral side. These observations suggest that not only the dopaminergic denervation of the striatum but also the imbalance in the activity of the thalamo cortical projection (reduced after 6-OHDA, augmented after IBO) are instrumental in determining the degree and direction of circling. Quantification of NPY immunoreactive neurons in striatum revealed a decrease in 6-OHDA lesioned rats after 3 days on the side contralateral to the lesion, an effect even more pronounced after 4 month's survival time. IBO-induced lesions of the SN had an opposite effect on NPY-immunoreactivity in the striatum: neuron counts were lower on the ipsi- than on the contralateral side. In addition, a time-dependent variation in total number of NPY-neurons was noted: during the early postoperative periods an increase, followed by a prolonged decrease to values below 50% of the controls after 4 months. Taken together, these results provide evidence that a dopaminergic deafferentation and its consequences on the nigro thalamo-cortical loop will determine NPY expression in the striatal interneurons. In particular, it is suggested that the number of striatal NPY-neurons and the imbalance in cortical activity are tightly coupled in terms of a negative correlation. PMID- 8420633 TI - Hippocampal damage does not impair instrumental appetitive conditioning with delayed reinforcement. AB - The present study evaluated the effects of bilateral hippocampal lesions on appetitive instrumental conditioning with delayed (5-s interval) reinforcement in rats. Acquisition of a bar press response was considerably slower than rates observed with immediate reinforcement; however, no significant differences between hippocampally lesioned and control groups were noted regarding training to criteria or ratio of responses to reinforcements. These results suggest that the hippocampus is not essential for the association of temporally discontinuous stimuli, and that deficits in other forms of associative learning, such as spatial cognition, must be mediated by the loss of other functions. Putative functions and underlying substrates are discussed for response modulation and sensory (cue relations) associations. PMID- 8420632 TI - Functional evidence that the angiotensin antagonist losartan crosses the blood brain barrier in the rat. AB - Losartan is a novel nonpeptidergic antagonist of angiotensin (ANG) II subtype 1 (AT1) receptors, which effectively lowers blood pressure in high-renin hypertensive rat and blocks the pressor response to systemic ANG II. It is well known that high densities of ANG II receptors exist in the hypothalamic paraventricular nucleus (PVN). In addition, activation of putative angiotensinergic afferents to the PVN originating in subfornical organ (SFO) elevates blood pressure and facilitates the activity of PVN neurons. We report here that systemic administration of losartan (3 mg/kg) significantly attenuates the pressor response to electrical stimulation of SFO. The excitatory responses of PVN neurons to SFO stimulation or local pressure microinjection of ANG II were also significantly inhibited in 58.8% and 88.9% of PVN cells, respectively, by intravenous administration of losartan. These pharmacological effects were rapid and reversible, and were accompanied by little change of basal arterial blood pressure or spontaneous neuronal activity. These observations suggest that systemic losartan crosses the blood-brain barrier (BBB) and acts at AT1 receptors within the PVN. PMID- 8420634 TI - Augmented neuronal activity in the hypothalamus of spontaneously hypertensive rats. AB - Previous studies indicate a tonic GABAergic inhibitory mechanism in the posterior hypothalamus (PH) contributes to modulating cardiovascular activity. Blockade of GABA receptors on neurons in this area elicits an increase in sympathetic discharge, arterial pressure, and heart rate. It has been proposed that a deficit in this inhibitory system may be responsible for the elevated pressure in the spontaneously hypertensive rat (SHR). The purpose of this study was to determine if the spontaneous neuronal activity in the posterior hypothalamus of spontaneously hypertensive rats differs from that of age-matched normotensive Wistar-Kyoto rats (WKY). Single unit, extracellular recordings of posterior hypothalamic neurons were performed on both in vivo and in vitro preparations. The spontaneous firing rate of posterior hypothalamic neurons in the anesthetized adult SHR was significantly higher (3.66 +/- 0.55 Hz) compared to that of the anesthetized adult WKY rat (2.11 +/- 0.29 Hz). Moreover, more of the neurons in the anesthetized SHR (38%) had a bursting discharge pattern than in the WKY (16%). In order to exclude inputs from peripheral receptors or other brain areas, an in vitro preparation was used. Neurons from both young and adult SHRs also had an increased spontaneous discharge rate and higher percentage of burster-type cells in the posterior hypothalamus compared to neurons from age-matched WKYs in the brain slice preparation. Both the in vivo and in vitro findings support the possibility that an elevated neuronal activity in the posterior hypothalamus, a known pressor area, of the SHR contributes to the development and/or maintenance of hypertension in this animal model. PMID- 8420635 TI - The acupuncture point and its connecting central pathway for producing acupuncture analgesia. AB - Characteristics of the acupuncture point in producing acupuncture analgesia (AA) were examined by the inhibition of noxious responses in the brain stem reticular formation, potentials, and neuronal activity in the dorsal periaqueductal central gray (D-PAG), and analgesia caused by low frequency stimulation of the acupuncture point. As a result, stimulation of the muscle beneath the acupuncture point was found to be effective in producing AA. AA measured by tail flick, vocalization, and writhing tests was abolished by hypophysectomy, and by antiserum of beta-endorphin administered into the 3rd ventricle. The pathway from the D-PAG to the anterior hypothalamus (AA-AH) in the AA afferent pathway from the acupuncture point to the pituitary gland was determined. The lateral hypothalamus, lateral septum, cingulate bundle, dorsal-hippocampus, and habenulo interpeduncular tract were found, in addition to regions previously found, to belong to the AA afferent pathway. A network of divergence and convergence in their rostral and caudal relations was observed. The AA afferent pathway diverges from the D-PAG, converges to the HP, and then projects to the AA-AH. PMID- 8420636 TI - GABA-like immunoreactivity in the gustatory zone of the nucleus of the solitary tract in the hamster: light and electron microscopic studies. AB - The distribution of GABA-like immunoreactive (GABA-LI) somata was studied in the gustatory zone of the nucleus of the solitary tract (NST) in the hamster in order to identify putative inhibitory circuitry in gustatory processing. Immunoreactive somata were located throughout the gustatory NST, in accordance to the distribution of large and small types of neurons as determined in previous morphometric studies. Consequently, GABA-LI somata were mostly found in the dorsal two-thirds of the gustatory zone. Such somata were mostly ovoid in shape and possessed somal areas that averaged 85.5 +/- 2.8 microns 2 (12.7 x 8.4 microns). A narrow range of somal areas (50-125 microns 2) suggested a single functional group. At the electron microscopic level, 18% of the neurons encountered were immunoreactive and their nuclei always possessed deeply invaginated boundaries. This morphological feature indicated that GABA-LI neurons are smaller members of the most common class of neurons within the gustatory NST. Because GABA is often implicated as the neurotransmitter of small inhibitory local circuit neurons, these findings indicate a possible inhibitory aspect to the processing of taste information at the level of the first relay in the brainstem. PMID- 8420638 TI - Genetic profiles of morphine-induced EEG, EEG power spectra, and behavior in two inbred rat strains. AB - The purpose of the present study was to examine and compare the effects of morphine at doses of 3, 10, and 30 mg/kg, IV, on EEG, EEG power spectra, and behavior in two inbred rat strains, Lewis and Fischer 344. Duration of morphine induced EEG slow-wave bursts and associated behavioral stupor was greater in Lewis rats. Latency to slow-wave sleep increased in a dose-related manner for both strains; differences were not significant between the two groups. The analog EEG signal was transformed by fast Fourier analysis; six power spectral quantities were examined: peak frequency, complexity, mobility, mean frequency, edge frequency, and total power. With the exception of peak frequency and edge frequency, all differed as a function of inbred rat strain. Regarding morphine dose, all spectral parameters differed except peak frequency. Factor analysis on morphine-induced EEG revealed a unique factor for each strain that was bipolar in nature and may be associated with the burst and interburst periods that occur in EEG after opiate administration. Genetic variability appears to play a role in the behavioral, EEG, and derived power spectral responses of both LEW and F344 inbred rat strains following acute morphine administration. These results may reflect differences in neurosensitivity and/or opioid receptor populations between Lewis and Fischer 344 inbred rat strains. PMID- 8420637 TI - Pituitary-adrenal function and hypothalamic beta-endorphin release in vitro following food deprivation. AB - Basal and dexamethasone-suppressed adrenal glucocorticoid secretion and hypothalamic beta-endorphin (BE) release in vitro were investigated in rats deprived of food for 24, 48, 72, and 96 h. Fasting for up to 48 h neither caused significant changes in serum corticosterone levels nor in the suppressive effect of dexamethasone. Food deprivation for 72-96 h resulted in increased basal serum corticosterone, diminished suppression by dexamethasone, and a significant involution of the thymus. Basal in vitro BE release from hypothalamic explants was significantly increased after the first day of food deprivation, and in vitro perifusion with corticotropin-releasing hormone (CRH) failed to enhance BE release further. With continuing food deprivation, basal BE release remained significantly greater than that from hypothalami originating from normally fed control rats. The stimulatory effect of CRH on BE release was only partially restored after 2 days of fasting. The results suggest that food deprivation for more than 2 days increases basal glucocorticoid secretion, and signs of impairment in hypothalamic-pituitary-adrenal regulation become apparent. These findings might be implicated in the pathogenetic mechanisms of endocrine dysregulation in diseases related to caloric reduction. PMID- 8420639 TI - Protein levels in the vitreous of rats with streptozotocin-induced diabetes mellitus. AB - The vitreous is a neural extracellular space separated from the blood-vascular compartment by the blood-retinal barrier. Study of the appearance of serum proteins in this space have been carried out in rats with streptozotocin-induced diabetes mellitus, a condition associated with barrier dysfunction. A vitreous sampling technique that avoids contamination with surrounding tissue was employed. In rats 1 month after administration of streptozotocin (fasting serum glucose > or = 375 mg/dl), significant increases in vitreous protein were observed in the absence of discernible eye pathology. Two-dimensional isoelectric focusing and SDS-polyacrylamide gel analysis of the soluble fraction demonstrated 85 polypeptides, 28 of whose electrophoretic positions coincided with positions of serum polypeptides. The remainder were unrelated to serum polypeptide loci. Overall patterns of soluble protein from the vitreous of streptozotocin-injected and normoglycemic-uninjected control animals were virtually identical. Results support a system for selective transfer for certain proteins into the extraneural vitreous space as suggested by Chen and Chen (6). PMID- 8420640 TI - Prenatal cocaine and cell development in rat brain regions: effects on ornithine decarboxylase and macromolecules. AB - Prenatal cocaine exposure has been shown to cause neurobehavioral abnormalities. To determine whether effects on basic patterns of cell development underlie these functional deficits, we examined the aftermath of acute and chronic cocaine exposure on ontogenetic patterns of ornithine decarboxylase (ODC), a key regulator of cell replication/differentiation, DNA synthesis as monitored by [3H]thymidine incorporation, and markers of cell number (DNA content) and cell size (protein/DNA ratio). Administration of 30 mg/kg SC of cocaine to pregnant rats on gestational day 20 resulted in acute increases of ODC throughout the brain. When the same dose of cocaine was given daily from gestational days 8 through 20, ODC elevations persisted into the neonatal period but disappeared by the middle of the first postnatal week. Although this treatment regimen retarded maternal weight gain, there was little or no effect on pup body or brain region weights. Similarly, minor changes in DNA synthesis were seen in two brain regions (forebrain, cerebellum), but DNA content was largely unaffected. Postnatal cell growth was significantly reduced in the forebrain, as evidenced by deficits in protein/DNA but, again, the magnitude of effect was quite small. Raising the daily dose of cocaine to 100 mg/kg resulted in significant maternal mortality and fetal resorptions in surviving dams. Shortening the treatment regimen to a 3-day period (gestational days 18 through 20) eliminated the effects on maternal weight gain and on postnatal pup brain region ODC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420641 TI - Continuing medical education at the annual meeting. PMID- 8420642 TI - Scary changes for physicians. PMID- 8420643 TI - "Best interests" standard not safe? PMID- 8420644 TI - Timing of the Medical Council of Canada clinical examination. PMID- 8420645 TI - Timing of the Medical Council of Canada clinical examination. PMID- 8420646 TI - Abortion debate continues. PMID- 8420647 TI - Abortion debate continues. PMID- 8420648 TI - How to diagnose and treat Lyme disease in children. PMID- 8420649 TI - Patient education about asthma. PMID- 8420650 TI - Patient education about asthma. PMID- 8420651 TI - Chiropractic manipulation. PMID- 8420652 TI - Can tamoxifen prevent breast cancer? PMID- 8420653 TI - The tobacco epidemic: how far have we come? PMID- 8420654 TI - Periodic health examination, 1993 update: 1. Primary prevention of child maltreatment. The Canadian Task Force on the Periodic Health Examination. PMID- 8420655 TI - The reuse of hemodialyzers: an assessment of safety and potential savings. AB - OBJECTIVE: To evaluate the safety and potential cost savings of hemodialyzer reuse. DATA SOURCES: All English and French articles published from 1960 to 1991 related to hemodialyzer reuse (retrieved through an Index Medicus and MEDLINE search [corrected]), the indexes of eight North American journals from 1960 onward, conference proceedings, association guidelines, and US and Canadian laws and regulations. RESULTS: For health care personnel the reuse of hemodialyzers did not entail any increased risk of infection or exposure to toxic substances if proper control measures were taken. For patients there was no evidence to suggest any excess risk of complications or death as long as precise and appropriate procedures are observed. The "first-use syndrome" can be prevented and should no longer be considered as a reason to favour reuse. A cost-minimization analysis indicated that five uses might save up to $3629 per patient yearly. Thus, the adoption of a policy of reuse in Canada for all eligible patients undergoing long term hemodialysis could result in direct savings of about $5.8 to $8.9 million per year. CONCLUSION: The health risks associated with hemodialyzer reuse can be reduced to acceptable levels through the rigorous observance of proper quality assurance and quality-control measures and the use of automated reconditioning equipment. Such a policy could achieve modest savings for the health care system. A decision to reuse should be formally adopted by the institution and accompanied by a precise definition of the standards of quality assurance and control. PMID- 8420656 TI - Trends in rates of admission to hospital and death from asthma among children and young adults in Canada during the 1980s. AB - OBJECTIVE: To update reports of increases in the rates of admission to hospital and death from asthma among children and young adults in Canada during the 1970s by examining data for the 1980s. DESIGN: Age-standardized rates were calculated from data for people less than 35 years of age at the time of death from asthma, bronchitis or other respiratory conditions (from 1980 through 1989) and at the time of admission to hospital for treatment of these diseases (from 1980 through 1988). Standardized mortality ratios were calculated with the death rate for Canada as the expected rate. SETTING: Data for all of Canada were examined by sex, age group and province. RESULTS: In contrast to sharp increases in the rate of death from asthma observed from 1970 through the early 1980s among Canadians less than 35 years of age, the rate showed no net change between 1980 and 1989; on average, there were 58 deaths in this age group annually. During the decade, the rates of death from asthma were three times higher in Saskatchewan and Alberta than in Newfoundland. The national rate of hospital admission/separation for asthma, however, increased greatly, though changes in the rate varied by province. Increases of over 90% were observed in Prince Edward Island and New Brunswick, whereas little overall change occurred in Newfoundland, Manitoba and Saskatchewan. The rate of hospital admission/separation for asthma was highest in Prince Edward Island and lowest in Manitoba and British Columbia. Although the rates of hospital admission/separation for asthma among boys aged less than 15 years of age were consistently 50% higher than those among girls of that age, the rate among people aged 15 through 34 years was twice as high among females as males. A slight decrease in the rates of death from respiratory conditions other than asthma was observed, together with a steady, fairly substantial decline in the rates of hospital admission/separation for these conditions. CONCLUSIONS: Whether there is any relation between increases in rates of admission to hospital for asthma and trends in the rates of death from asthma during the decade will require further study. PMID- 8420657 TI - [Prevalence of cervical Chlamydia trachomatis infection in a female population seeking contraception counseling]. AB - OBJECTIVES: To determine the prevalence and risk indicators of cervical infection due to Chlamydia trachomatis among female patients consulting for contraception and to evaluate an enzyme immunoassay for the detection of C. trachomatis in this setting. DESIGN: Prevalence study. Endocervical specimens were analysed by means of culture and enzyme immunoassay. C. trachomatis infection was diagnosed through culture. SETTING: A hospital family planning clinic in Trois-Rivieres, Que. SUBJECTS: All 533 female patients who consulted for contraception between November 1986 and March 1988. Results of culture were available for 495 patients. MAIN OUTCOME MEASURE: Demographic, epidemiologic and clinical information was collected by means of a standard questionnaire and a gynecologic examination. MAIN RESULTS: The prevalence rate of chlamydial infection was 9% (45/495). Enzyme immunoassay detected 37 (82%) of the infections. The mean age of the patients was 19.8 years, and 98% of the infections were diagnosed in those aged 25 years or less. The variables significantly associated with C. trachomatis infection were having more than one sexual partner in the preceding year (odds ratio [OR] 2.9; 95% confidence limits [CL] 1.7 and 5.0) and having more than one partner in the preceding 3 months (OR 2.3; 95% CL 1.2 and 4.3). These two indicators would have detected 58% and 22% of the infections respectively. CONCLUSIONS: Screening for C. trachomatis infection by means of enzyme immunoassay should be proposed to all female patients aged 25 years or less consulting for contraception in our clinic. Such screening may prove to be an effective preventive measure in other similar clinical settings. PMID- 8420658 TI - Cigarette smoking: a clinical and public health challenge. Part II. The Public Health Challenge. 1962. PMID- 8420659 TI - Chance and the blood count. 1934. PMID- 8420660 TI - National Non-Smoking Week a time to consider battles won, victories needed: CMA. PMID- 8420661 TI - Doctors need to be more vocal in opposing smoking, AMA representative tells Canadian MDs. PMID- 8420662 TI - Breast cancer activists call for more research, more compassion. PMID- 8420664 TI - Cairo residents find signs of hope in wake of city's worst natural disaster. PMID- 8420663 TI - PEI's video-gambling machines creating an addiction problem, island MDs warn. PMID- 8420665 TI - BC report on sexual abuse by MDs "tough and fair," college past president says. PMID- 8420666 TI - New native healing centre in Toronto opens eyes of non-native MDs who work there. PMID- 8420668 TI - Report may sound death knell for some of London's historic hospitals. PMID- 8420667 TI - Female circumcision: when medical ethics confronts cultural values. PMID- 8420669 TI - The HIV-positive health care worker and testing for HIV. PMID- 8420670 TI - Older women as the focus for research and treatment of ovarian cancer. An overview for the National Institute on Aging, National Cancer Institute, and American Cancer Society Multidisciplinary Working Conference. AB - Ovarian cancer disproportionately affects women 65 years of age and older who are likely to have concomitant changes in physical ability, physiological functioning, and other chronic conditions associated with advancing age. The National Institute on Aging, National Cancer Institute, and the American Cancer Society cosponsored a multidisciplinary working conference, "Perspectives on Ovarian Cancer in Older-Aged Women: Current Knowledge and Recommendations for Research," at the National Institutes of Health on November 20-21, 1991 to confront the age-related aspects of ovarian cancer in epidemiology, etiology, clinical investigations, and patient management. Conference participants devoted attention to such special topics as drug resistance, dose intensity, patterns of care, and screening potential for ovarian cancer. After exploring these areas with existing data, the task was then to generate recommendations for research and practice. The scope of the conference and an introduction to the proceedings are presented in this paper. PMID- 8420671 TI - Ovarian cancer. Age contrasts in incidence, histology, disease stage at diagnosis, and mortality. AB - BACKGROUND: Age comparisons for incidence, histology, disease stage at initial diagnosis, and mortality of more than 20,000 ovarian cancer patients diagnosed between 1973-1987 are the focus of this descriptive epidemiologic study. This paper highlights key issues and concerns regarding ovarian cancer in women 65 years and older as a frame of reference for the proceedings of the working conference, "Perspectives on Ovarian Cancer in Older-Aged Women," co-sponsored by the National Institute on Aging, National Cancer Institute, and American Cancer Society held at the National Institutes of Health, November, 1991. METHODS: Data are from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) Program and the National Center for Health Statistics. The SEER Program database represents approximately 9.6% of the U.S. population. RESULTS: Ovarian cancer affects women in the age group 65 years and older more frequently than younger women. More than 48% of all ovarian cancers occur in women in this age group. Age-adjusted rates increase as age advances, peaking at 54.0 per 100,000 in the age group 75-79 years. Time trends also indicate increases in age specific incidence rates. This malignancy takes its toll in mortality in women 65 years and older with 64% of all deaths due to this neoplasm (in 1989). Moreover, older women are more likely to be initially diagnosed with advanced disease. CONCLUSIONS: Important questions about ovarian cancer in older-aged women need urgent attention from the research community. New strategies for diagnostic leads have to be developed for older women. PMID- 8420672 TI - Ovarian cancer. Survival and treatment differences by age. AB - BACKGROUND: Cancer of the ovary is a disease of older American women with an incidence rate of 9.4 per 100,000 for those under 65 compared to 54.8 per 100,000 for those 65 years of age and over. METHODS: Over 22,000 women were diagnosed with ovarian cancer between 1973 and 1987 within the Surveillance, Epidemiology and End Results (SEER) Program of the National Cancer Institute. SEER is a population-based program that covers nearly 10% of the U.S. population for cancer incidence and survival. RESULTS: Ovarian cancer survival rates vary dramatically by stage. Within stage, however, differences are noted in survival by age, with younger women surviving better than older women even after adjustment for the general life expectancy of each age group (relative survival). For Stages III-IV disease, women under 45 years of age have a 5-year relative survival rate of over 45% compared to only 8% for those 85 years of age and over. Between 1973-1977 and 1983-1987, the treatment for Stages III-IV disease has changed. For all age groups, there were sharp increases in the percentage having surgery and chemotherapy/hormonal therapy and decreases in those having surgery and radiation as part of the first course of therapy. Over 40% of women 85 years of age and over did not receive any definitive treatment according to the hospital medical record. In 1983-1987, younger women received more combination therapy (surgery with chemotherapy/hormonal therapy) versus older women who received more single modalities such as surgery only or chemotherapy/hormonal therapy only. CONCLUSIONS: Older women with ovarian cancer are treated less aggressively than their younger counterparts and have poorer survival rates. PMID- 8420673 TI - The biology of ovarian cancer development. AB - BACKGROUND: In theory, all the cell types that comprise the human ovary have the potential for malignant transformation. The vast majority of malignant ovarian tumors in the human, however, arise from the ovarian surface epithelium. These cells have important functions during reproductive life; they contribute to follicular rupture and by cell division repair the wound that accompanies ovulation. There has been much speculation that the rapid cycles of cell division associated with wound repair contribute significantly to the development of ovarian cancer. Such speculation is based on the observation that ovarian cancer occurs most frequently at the end of a woman's reproductive life and is associated with nulliparity. It is of potential significance that, unlike most epithelia, these cells are not replaced through replenishment stem cells with the development of one end-stage cell and one cell with continued growth potential. Rather, the division of an ovarian surface epithelial cell yields two daughter cells with equal potential for subsequent growth. Thus, all potential mutations as they accumulate are passed on to near-exponentially expanding subsequent generations of cells that can acquire additional mutations that could confer the malignant phenotype. METHODS: We have developed a model to test the hypothesis that repeated cell division by ovarian surface epithelial cells contributes to development of malignancy. In this model, rat ovarian surface epithelial cells are isolated and subjected in vitro to repetitious cell division to mimic in a simple way growth of the surface epithelium in vivo. RESULTS: These cells develop a malignant phenotype based on loss of contact inhibition, the ability for substrate independent growth, tumorigenicity in athymic mice, and cytogenetic changes. CONCLUSIONS: Our data support the involvement of tumor suppressor genes in the development of ovarian cancer. PMID- 8420674 TI - Age contrast in ovarian pathology. AB - BACKGROUND. Ovarian cancer accounts for approximately 23% of all gynecologic tumors and is the most common fatal gynecologic malignancy. These tumors can occur in women of all ages, but there are differences in the histologic types during various decades of life. CONCLUSIONS. During infancy and childhood, the predominant type of neoplasms are those of germ cell origin, such as teratomas, dysgerminomas, and endodermal sinus tumors. In adults, epithelial neoplasms, or tumors that originate from the epithelium that covers the ovarian surface, are the most common, accounting for almost 85% of all neoplasms after the age of 50 years. The peak incidence of benign epithelial tumors occurs between the ages of 20 and 40 years. Young women (30 to 40 years of age) are frequently affected by the so-called "tumors of low malignant potential," which have excellent prognosis. Older women, on the other hand, usually have the most aggressive forms of ovarian cancer, present with advanced disease, and have a dismal prognosis. PMID- 8420675 TI - Growth regulation and transformation of ovarian epithelium. AB - The discovery of peptide growth factors and cancer-causing genes (oncogenes and tumor-suppressor genes) has provided us with the exciting opportunity to begin to understand the molecular pathology of human ovarian cancer. Activation of several genes, including HER-2/neu, myc, ras, and p53 have been described in some ovarian cancers. In addition, some protooncogenes such as the epidermal growth factor receptor (erbB) and the M-CSF receptor (fms) are expressed along with the respective ligands (peptide growth factors) in some ovarian cancers. Although the studies reviewed in this paper represent a promising beginning, we remain far from a comprehensive understanding of growth regulation and transformation of human ovarian epithelium. PMID- 8420676 TI - Aging issues in invasion and metastasis. Fertile ground for investigation. AB - BACKGROUND: Ovarian cancer is a disease of predominantly older women and one in which metastases occur early and extensively. The unusual pattern of local metastasis is fertile ground for laboratory and clinical investigation. METHODS: Tools are now available with which to study the complex cascade of invasion and metastasis as it underlies invasive ovarian cancer. The metastatic cascade involves five repetitive steps: angiogenesis, adhesion to the vascular basement membrane, local proteolysis, migration into and out of the vasculature, and proliferation at the secondary sites. Both in vivo and in vitro models of ovarian cancer are available; however, there are no systems targeted to understanding age related differences in ovarian cancer biology. RESULTS: Progress in investigation of the biology of ovarian cancer has led to new diagnostic and therapeutic leads, including the use of adhesion receptors, protease secretion, and stimulation of tumor cell migration as potential markers and the identification of a new anti cancer agent, CAI (NSC609974). CONCLUSIONS: CAI inhibits signal transduction pathways important in the regulation and activation of metastasis and proliferation. A phase I study has begun accrual at the National Cancer Institute; the protocol will contain no upper age limit. PMID- 8420677 TI - Personal characteristics relating to risk of invasive epithelial ovarian cancer in older women in the United States. AB - BACKGROUND: The ovarian cancer incidence rate is higher for women over 55 years of age than for younger women, but it increases less rapidly with age after age 55 years than before. This slower rate of increase suggests that the postmenopausal ovary may be less exposed or less susceptible than the functioning one to endogenous or exogenous carcinogens. For example, it is not exposed to mitotic stimuli at follicle rupture, which may increase the likelihood of malignancy. METHODS: Findings from a collaborative analysis of 12 U.S. case control studies are used to examine associations between invasive epithelial ovarian cancer and certain reproductive and hormonal characteristics, and to determine if those associations change as women age. RESULTS: Ovarian cancer risk reductions were associated with pregnancy (regardless of outcome), lactation, and oral contraceptive use. The percentage risk reduction associated with pregnancy was smaller for older women than younger women, although the absolute magnitude of the risk reduction increased with age. In contrast, the percentage risk reduction associated with oral contraceptive use was greater for older women. The total duration of ovulation was strongly associated with increased risk in women under 55 years of age, but not in older women. CONCLUSIONS: The greater protection to older women associated with oral contraceptive use suggests that the early high-potency contraceptive formulations used by these women may have been more protective than recent ones. The sparsity of strong risk factors in older women emphasizes the need for sensitive and specific markers for early detection of ovarian malignancy in this age group. PMID- 8420678 TI - Genetic epidemiology of epithelial ovarian cancer. AB - BACKGROUND: Aside from age, family history is the strongest predictor of ovarian cancer risk. Genetic components of risk for ovarian cancer have been evaluated by a number of designs, including case-control studies of family history and other risk factors, segregation and genetic linkage studies, and studies of biomarkers and tumor-specific cytogenetic abnormalities. METHODS: Data were extracted from all available case-control studies that included family history. Cytogenetic, biomarker, segregation, analytic, and genetic linkage studies were reviewed. RESULTS: Family history of ovarian cancer confers a 3.6-fold increased risk for this disease. Segregation studies of breast and ovarian cancer in five large families were consistent with dominant inheritance. Low levels of alpha-L fucosidase confer mildly increased risk for ovarian cancer. Low galactose-1 phosphate uridyl transferase and type A blood group may increase risk for ovarian cancer. Cytogenetic and oncogene studies have identified regions that may be important in tumorigenesis and metastasis, but discriminating between early and late changes is difficult from these studies. Presence of a genetic susceptibility locus for breast and ovarian cancer has been confirmed on chromosome 17q21. CONCLUSIONS: Family history is an important predictor of ovarian cancer risk. In rare families, a specific dominantly acting gene can be identified, but in the vast majority of familial ovarian cancers the underlying mechanism remains unclear. Specific studies are needed for women with a family history of ovarian cancer because evidence suggests modification of the effects of oral contraceptive use and reproductive patterns in this population of women. PMID- 8420679 TI - Hereditary ovarian cancer. Heterogeneity in age at onset. AB - BACKGROUND: Hereditary ovarian cancer (HOC) is heterogeneous, with at least three distinctive syndromes, namely, hereditary site-specific ovarian cancer, hereditary breast-ovarian cancer (HBOC) syndrome, and Lynch syndrome II. Ovarian cancer, in accord with virtually all varieties of adult onset cancer, displays an increasing incidence with advancing age; however, it shows an earlier age of onset in hereditary settings. METHODS: Detailed medical and pathology studies were performed on extended ovarian cancer-prone pedigrees, with special attention given to age at ovarian cancer onset. RESULTS: The age of onset of ovarian cancer is heterogeneous, wherein the average age of onset in HBOC is 52 years, in hereditary site-specific ovarian cancer it is 49 years, and in the Lynch syndrome II it is 45 years, in contrast to its occurrence in the general population, at an average age of 59 years. CONCLUSIONS: These differences are important for the initiation of surveillance and management strategies. Age of onset of ovarian cancer differences in these several hereditary subsets are less striking than they are in the case of other integral forms of cancer in the respective syndromes, such as the breast in the HBOC syndrome. In addition, the phenomenon of extremely early age of onset of ovarian cancer occurs infrequently in HOC when compared to other forms of cancer, such as the breast in HBOC or the colon in Lynch syndrome II. Knowledge about age of onset heterogeneity in HOC may harbor important clues about etiology, pathogenesis, and cancer control. PMID- 8420680 TI - Familial ovarian cancer. A report of 658 families from the Gilda Radner Familial Ovarian Cancer Registry 1981-1991. AB - BACKGROUND: Because of the small number of cases (five) reported between 1929 and 1969 and a significant increase reported in the decade of the 1970s, the Familial Ovarian Cancer Registry was established in 1981 to study the occurrence of familial ovarian cancer in the United States. METHODS: Any woman (with or without ovarian cancer herself) who contacted the Registry and demonstrated a familial history of ovarian cancer was added to the Registry as an index case. RESULTS: From 1981 through May 31, 1991, 658 families for a total of 1568 cases of familial ovarian cancer were accessioned into the Registry. Of the 219 mothers and 251 daughters with familial ovarian cancer, the mean (58.5 years) and median (57.0 years) age at diagnosis of the mothers was significantly older than the mean (49.8 years) and median (49.0 years) ages of their daughters with ovarian cancer. Significantly more index cases without ovarian cancer had used oral contraceptives as compared to index cases with ovarian cancer (P < 0.001). Significantly more index cases with ovarian cancer used other estrogens as compared to index cases without ovarian cancer (P < 0.001). The Registry cases exhibited a higher proportion of serous adenocarcinoma, poorly differentiated adenocarcinoma, borderline carcinoma, and gonadoblastoma as compared to the 1978 SEER data. Mother and a minimum of one daughter was the most common relationship and was represented in 49.5% of the families with familial ovarian cancer. Sister sister relationships were the second most frequent and accounted for 38.5% of the 658 families. CONCLUSIONS: Familial ovarian cancer occurs most frequently in mother-daughter relationships followed by sister-sister and appears to be an autosomal dominant inheritance with variable penetrance. PMID- 8420681 TI - A National Cancer Institute sponsored screening trial for prostatic, lung, colorectal, and ovarian cancers. AB - BACKGROUND. Ovarian cancer is the fifth most common cause of cancer-related death in American women. The median age at diagnosis is about 62 years; incidence rises rapidly after age 60. Pelvic examination has been the primary method for detection of ovarian carcinoma. It is insensitive for the detection of early disease, however: most women present with disease beyond the pelvis (Stages III and IV) and are not curable with existing techniques. Two new technologies may be useful as screening tools for earlier detection of ovarian cancer. CA 125 is an antigenic determinant expressed on an ovarian cancer cell line. Transvaginal ultrasound (TVUS) images the ovaries from within the vagina and can be performed by a technician in about 10 minutes. In small preoperative studies of women with ovarian masses, serum CA 125 levels have been elevated (typically above 35 U/ml) in over two-thirds of cases and in up to 50% of Stage I cases. The test is not absolutely specific: elevations have been reported with pregnancy, endometriosis, menstruation, benign ovarian tumors, and with cancers of the breast, colon, pancreas, lung, stomach, and liver. Nevertheless, the specificity of CA 125 in postmenopausal women has been reported at about 95% or more. TVUS provides higher resolving power for ovarian abnormalities than transabdominal ultrasound or physical examination; however, experience with it is limited. CA 125 and TVUS may be complementary. CONCLUSIONS. For these reasons, the National Cancer Institute is planning a randomized trial of all three tests versus routine medical care in women of ages 60-74 years. This is part of a larger trial to determine the efficacy of screening for lung, colorectal, and ovarian cancers in women, and for lung, colorectal, and prostatic cancers in men. Seventy-four thousand women will be randomized in the study. PMID- 8420682 TI - Localized ovarian cancer in the elderly. The Gynecologic Oncology Group experience. AB - BACKGROUND: The appropriate therapy for patients with localized (FIGO Stage I and II) ovarian cancer has been poorly defined for all age groups and particularly for the elderly. Few prospective randomized comparisons of adjuvant therapy after careful surgical staging have been performed. The Gynecologic Oncology Group (GOG) has performed a series of trials testing adjuvant treatment in carefully staged patients with early-stage ovarian cancer. Early trials included few elderly patients but the most recent trial (GOG 95) included 18% over the age of 65 years. METHODS: Comprehensive surgical staging defined by protocol is performed before randomization. Patients with predefined stages and histologies are included and the patients are randomized prospectively to receive either intraperitoneal phosphorus-32 or three monthly cycles of cyclophosphamide and cisplatin. Assessment of the value of this adjuvant therapy will depend on survival, disease-free survival, and relapse pattern differences between the two adjuvant therapies. RESULTS: This is an ongoing clinical trial and insufficient numbers of patients have been randomized for definitive conclusions. There have been seven recurrences on both arms of the trial with a median time to recurrence of 14 months. There currently are no significant age differences between relapsed patients and disease-free patients. At this point, 12 elderly patients have been randomized to each of the arms of therapy. CONCLUSIONS: Although no apparent survival differences exist for elderly patients in the most recent adjuvant chemotherapy trial of early ovarian cancer, the number of patients with cancer randomized and follow-up are insufficient to establish such a difference. Currently there is no evidence that elderly patients display a significant difference in relapse frequency or pattern. PMID- 8420683 TI - Age as a prognostic factor in ovarian carcinoma. The Gynecologic Oncology Group experience. AB - BACKGROUND: The Gynecologic Oncology Group (GOG) has completed six major randomized trials in advanced ovarian carcinoma over the 15-year period between 1976 and 1990. This large database of 2123 patients provides a well-studied patient population with which to examine the importance of age as a prognostic factor. METHODS: The 2123 patients studied in the six GOG trials were analyzed as a group to determine important prognostic factors. Further analyses were then conducted to examine outcome by decade of life from younger than 40 years old to 70 years old and older and to evaluate the interaction of age with other significant prognostic variables. RESULTS: Three major prognostic factors were identified as exerting an influence on patient outcome in the overall patient population: age, volume of residual disease, and performance status. With regard to the effect of age, patients older than 69 years of age exhibited significantly poorer survival than those younger, even after correction for stage, residual disease, and performance status. This was not altered by variations in drugs, doses, and schedules; but there was no evidence that older patients tolerated intensive schedules less well than younger patients. CONCLUSIONS: Two practical conclusions result from this analysis. First, there is no evidence that modification of the drugs and schedules that make up the regimens used can overcome the adverse effect of older age. Second, age does not adversely affect the dose intensity that can be achieved; hence, age in itself is not reason to withhold or attenuate intensive chemotherapy, particularly in light of the fact that older patients have a poorer prognosis. PMID- 8420684 TI - Treatment of ovarian cancer in elderly women. Mayo Clinic-North Central Cancer Treatment Group studies. AB - BACKGROUND: With a 25% reduction in ovarian cancer mortality rate between 1973 and 1988 among younger American women, the overall control of ovarian cancer appeared to be improving. Unfortunately, American women older than 65 years of age experienced a 16% increase in mortality from this disease during that same interval. We examined our therapeutic outcomes following systemic chemotherapy administered to women of various ages accepted for treatment on phase III Mayo Clinic and North Central Cancer Treatment Group protocols between 1974 and 1988. METHODS: Three randomized studies of chemotherapy for Stage III and IV epithelial ovarian carcinoma were analyzed for possible effects of age on the results of treatment. All of the patients, regardless of age, had been enrolled and treated according to standard accession and dosage adjustment criteria. RESULTS: Among our 383 patients, 107 (28%) were 65 years of age or older. Although the elderly women tolerated our five different chemotherapy regimens nearly as well as did the younger patients, we found that progressively greater dose reductions were required for treatment continuation with advancing age between groups aged 44 years or younger, 45-64 years, and 65 years and older. When nonserous histology, Stage IV, ECOG performance status above 0, tumor grade above 1, and extent of residual disease greater than 2 cm were considered, our Cox model analysis yielded no firm evidence that age 65 years and older per se (P = 0.58) was a negative prognostic factor for survival. CONCLUSIONS: Elderly women eligible for randomized clinical trials tolerated Stage III and IV epithelial ovarian carcinoma and its chemotherapy nearly as well as did younger women. Among this population of women accepted for study in our three clinical trials, age 65 years and older per se was not proven to be a negative prognostic factor in our multivariate analysis. PMID- 8420685 TI - Analysis of patient age as an independent prognostic factor for survival in a phase III study of cisplatin-cyclophosphamide versus carboplatin-cyclophosphamide in stages III (suboptimal) and IV ovarian cancer. A Southwest Oncology Group study. AB - BACKGROUND: The purpose of this study was to evaluate the effect of age (i.e., less than 65 years or 65 years of age and older) on survival in a recently completed phase III Southwest Oncology Group study in ovarian cancer patients. METHODS: Multivariate and univariate regression analyses were used to identify independent prognostic factors of survival in 342 patients with previously untreated Stage III (suboptimal) or Stage IV ovarian cancer who participated in a randomized, phase III study of intravenous (I.V.) carboplatin 300 mg/m2 plus I.V. cyclophosphamide 600 mg/m2 versus I.V. cisplatin 100 mg/m2 plus I.V. cyclophosphamide 600 mg/m2 every 4 weeks for six courses. RESULTS: Multivariate regression analysis showed the following variables to be independent prognostic factors of survival: age (P = 0.04); performance status (P = 0.004); disease stage (P = 0.03); and race (P = 0.05). Patients under 65 years of age survived significantly longer than those 65 years or older, especially patients with a performance status of 2. Patients with a baseline performance status of 0-1 survived longer than patients with a performance status of 2, and Stage III patients longer than those with Stage IV disease. An unexpected finding was that white patients survived significantly longer than black patients, regardless of age, performance status, or stage of disease. Carboplatin-cyclophosphamide treated patients experienced similar survival and significantly less nausea and emesis, renal toxicity, hearing loss, tinnitus, neuromuscular toxicities, and alopecia. CONCLUSIONS: Ovarian cancer patients with advanced disease who are 65 years of age or older and/or with a performance status of 2 have significantly decreased survival compared to their younger and/or less debilitated counterparts. Carboplatin-cyclophosphamide is the recommended treatment (rather than cisplatin-cyclophosphamide), especially for older or debilitated patients because it is associated with less toxicity and similar survival. PMID- 8420686 TI - Patterns of care in carcinoma of the ovary. AB - BACKGROUND: During an epidemiologic study of women with ovarian cancer, the type of incision, intraoperative evaluation of the extent of disease, frequency of mistaken diagnosis, and ovarian cancer occurring in women who had a prior hysterectomy were reviewed. METHODS: The thoroughness of intraoperative evaluation of the extent of disease as well as the type of incision in 291 women with ovarian cancer was investigated. Within this group 41 women had undergone hysterectomy with retention of one or both ovaries before the diagnosis of ovarian cancer. An additional 43 women were discovered not to have primary carcinoma of the ovary. RESULTS: Seventy-one percent of the women with transverse incisions had incomplete intraoperative evaluations of the extent of disease compared with 42% of the vertical incision group. Medical record documentation revealed that 97% of the cases operated on by gynecologic oncologist had complete staging evaluations performed, but only 52% and 35% of cases operated on by obstetricians/gynecologists and general surgeons, respectively, were evaluated adequately. Of the 43 women discovered not to have primary carcinoma of the ovary, 5% had benign ovarian tumors, 3% had primary carcinoma of the peritoneum, and 5% had primary carcinoma of the intestines. Women who had their ovaries retained at hysterectomy and later had invasive ovarian cancer had approximately an 80% mortality. CONCLUSIONS: The mistaken diagnosis of carcinoma of the ovary can be reduced if surgeons and pathologists improve communications between themselves. The greater use of preoperative second opinions by gynecologic oncologists in women suspected of ovarian cancer should enhance quality of care. PMID- 8420687 TI - Epithelial ovarian cancer in the elderly. The Memorial Sloan-Kettering Cancer Center experience. AB - BACKGROUND: There is evidence of differences between older (> or = 65 years of age) and younger (< 65 years of age) women with ovarian cancer. METHODS: To evaluate differences in the care of older versus younger patients with ovarian cancer, a retrospective review was conducted of the records of 146 patients who had their initial surgery for ovarian cancer at the Memorial Sloan-Kettering Cancer Center from January 1987-January 1991. RESULTS: There was a significant difference in stage at presentation between the older (48 patients) and younger (98 patients) populations (Stage I/II: older 10%; younger 25%; P < 0.05). Although there was no statistically significant difference in the distributions of patients according to tumor grade between the two groups, 10% of the younger patients had ovarian cancer of low malignant potential, compared to only 2% of older patients (P < 0.1). Forty-six percent of the younger patients entered an intensive initial chemotherapy trial compared to only 17% of the older patients (P < 0.001), principally due to comorbid medical conditions (e.g., heart disease). Finally, whereas the relative number of younger to older patients with ovarian cancer undergoing initial surgery at this institution was 2:1, the relative number of totally new patients to the service (including referral for initial treatment or salvage programs) was 4:1 (P < 0.001), suggesting that older patients with ovarian cancer are less likely to be referred for secondary experimental programs than for initial treatment. CONCLUSIONS: These data suggest that there are major differences between the presentation and treatment strategies of older compared to younger patients with ovarian cancer. PMID- 8420688 TI - Age contrasts in patients with advanced epithelial ovarian cancer. The M.D. Anderson Cancer Center experience. AB - BACKGROUND: This study reviews the experience at the University of Texas M.D. Anderson Cancer Center with elderly patients treated for advanced epithelial ovarian cancer with cisplatin-based combination chemotherapy. METHODS: From 1978 through 1988, 215 patients with Stage III or IV, Grade 2 or 3 epithelial ovarian cancer were entered onto one of three consecutive trials involving cisplatin based combination chemotherapy. The treatment plans were as follows: Trial 1: 12 cycles of cisplatin-melphalan (75 patients); Trial 2: 12 cycles of cisplatin cyclophosphamide (49 patients); and Trial 3: 6 cycles of cisplatin cyclophosphamide (91 patients). End-points of analysis included the effect of age on other prognostic factors, clinical response rate, surgical response rate, toxicity, progression-free survival, and survival. RESULTS: Of the entire study group, 57 patients (27%) were 65 years of age and older at diagnosis. There was no effect of age on FIGO stage or histologic grade distribution, but 61% of patients younger than 65 years of age had residual tumors less than or equal to 2 cm compared with only 33% of patients 65 years of age and older (P = 0.00027). There were no differences between the two age groups with respect to response rates, recurrence rate after negative second-look surgery, toxicity, or progression-free survival. Patients in the younger than 65 years of age group, however, had a significantly longer median survival time than those 65 years of age and older (30 versus 19 months, respectively) (P = 0.0038). CONCLUSIONS: This analysis suggests that elderly patients are more likely to begin chemotherapy with bulky residual disease and to have a significantly shorter survival time than their younger counterparts. PMID- 8420689 TI - Mechanisms of drug resistance in ovarian cancer. AB - Alkylating agents, natural products and platinum complexes are the primary chemotherapeutic agents used in the treatment of patients with ovarian cancer. Resistance frequently develops to all three classes of drugs and can be functionally separated into distinct biochemical pathways: (1) relative dose intensity plays a role in resistance to platinum complexes and to a lesser degree with alkylating agents; (2) induction of the membrane P-170 glycoprotein confers resistance to natural products and due to the potential usefulness of Taxol (a natural product extracted from the bark of yew trees), this mechanism of resistance may become more clinically relevant in the future; (3) increased levels of cellular glutathione (GSH) and glutathione S-transferases are important in the detoxification of alkylating agents and platinum complexes; and (4) increased DNA repair also is characteristic of resistance to platinum complexes and alkylating agents. Clinical trials have been initiated with agents that may inhibit the biochemical mechanisms of acquired drug resistance. Clinical trials are already in progress with alkylating agents combined with inhibition of GSH biosynthesis (i.e., buthionine sulfoximine) or enzymatic inhibitors of glutathione S-transferase activity (i.e., ethacrynic acid). Furthermore, the combination of aphidicolin, an inhibitor of DNA repair, together with platinum complexes also soon will be clinically tested based on promising results in preclinical models of ovarian cancer. Ovarian cancer is a disease of the elderly. Advances in the pharmacology of platinum compounds and in our understanding of the mechanisms of drug resistance should permit these patients to receive increasingly more effective chemotherapy. PMID- 8420690 TI - Dose intensity analysis of high-dose carboplatin in refractory ovarian carcinoma relative to age. AB - BACKGROUND: Cisplatin and carboplatin are essential to management of advanced stage epithelial ovarian carcinoma. No published data exist regarding carboplatin dose intensity importance in ovarian cancer, nor are there data for the effect of age on the ability to deliver dose-intensive carboplatin. METHODS: Retrospective dose intensity analyses were performed for 93 patients with advanced-stage refractory ovarian carcinoma, who received single-agent, high-dose carboplatin chemotherapy (800 mg/m2/35 days of 160 mg/m2/week). These patients had been treated on one of three high-dose carboplatin studies conducted in the Medicine Branch of the National Cancer Institute during the 1980s. Eligibility criteria required age greater than or equal to 18 years, good end organ function, and good performance status. RESULTS: Patients 60 years of age or older comprised 33% of the cohort and patients 70 years of age or older comprised 8% of the cohort. Administered dose intensity of carboplatin did not differ among age groups. Patients of age < or = 39 years received a dose intensity of 136 +/- 28 mg/m2/week, those of age 40-59 years received 129 +/- 33, those of age > or = 60 years received 134 +/- 24, and those of age > or = 70 years received 129 +/- 19. Further, the administered carboplatin dose intensity did not differ among clinical response groups. Dose intensity in complete responders was 138 +/- 18 mg/m2/week; in partial responders, 121 +/- 29; and in nonresponders, 134 +/- 30. The age distribution of responders matched the age distribution of the cohort. CONCLUSIONS: These data demonstrate that elderly patients with good end organ function and good performance status tolerate the same level of dose-intensive platinum therapy as younger patients. Age alone should not be a determinant for carboplatin dose modification in the treatment of ovarian carcinoma. PMID- 8420691 TI - Transcranial magnetic stimulation (TMS) of the brain in patients with mesiotemporal epileptic foci. AB - Transcranial magnetic stimulation (TMS) of the human brain is mainly used for the diagnosis of diseases with disturbed central motor conduction. Recent studies revealed controversial results concerning the possibility of a TMS-induced specific activation of epileptogenic foci in patients with localization-related epilepsies, which would make TMS an additional diagnostic tool for the presurgical localization of the primary epileptogenic zone. We applied TMS to 19 patients with complex-partial seizures and investigated its effects and safety. In 12 patients we performed TMS during scalp electroencephalogram (EEG) recordings. The remaining 7 patients with localization-related epilepsies of mesiobasal limbic seizure origin underwent EEG with additionally implanted foramen-ovale-electrodes (FOE). We did not notice any significant spike activation and even observed bilateral reduction of epileptic activity in some patients. On the contrary, hyperventilation induced a marked activation of the epileptic focus. Our findings support that TMS is safe since adverse effects did not occur. However, due to possible safety hazards, TMS in epileptic patients still requires cautious application until more data will be available. PMID- 8420692 TI - Frontal intermittent delta activity in schizophrenic patients receiving antipsychotic drugs. AB - FIRDA was found in 14 EEGs recorded from 338 schizophrenic patients receiving antipsychotic drugs, although they showed no FIRDA in the baseline EEG. The daily dose of antipsychotic drugs when FIRDA was found was larger than when FIRDA disappeared. FIRDA was assumed to be induced by relatively high doses of antipsychotic drugs. Thus, the effect of antipsychotic drugs should be added to the list of differential diagnoses associated with FIRDA. FIRDA did not correlate with the baseline psychopathology of schizophrenic patients. Patients without FIRDA tended to respond poorly to antipsychotic drugs in terms of negative symptoms. Improvement of positive symptoms in FIRDA patients was not as remarkable as in patients without FIRDA. PMID- 8420693 TI - Visual event related potentials after methylphenidate and sodium valproate in children with attention deficit hyperactivity disorder. AB - We divided a group of Attention Deficit Hyperactivity Disorder (ADHD) boys into two groups determined by the results of EEG and visual ERP tests--those who had excessive EEG background slowing, EEG epileptiform activity or abnormally high amplitude ERP waves (the "E-ADD" group), and those with normal EEG and ERP (the "ADD" group). The subjects of each group were then given either a dose of methylphenidate (Ritalin, MPH), sodium valproate (Depakene, VPA) or placebo, and repeated visual ERPs one hour later. Following both medications there was a significant suppression of the amplitude of the N3, the "slow negative" wave of the ERP, in the E-ADD group but not in the ADD group. The effect of VPA and MPH in ADHD children may be related to the electrophysiological (EEG, ERP) background. Both medications may have a similar effect on arousal processes in ADHD. PMID- 8420694 TI - Electroencephalography in childhood conduct and behavior disorders. AB - The pathophysiology of behavior disorders in children is controversial. In particular, the relationship of episodic behavior disturbance to epilepsy and chronic behavior problems to subclinical neurologic disorder has been debated. It has been suggested that EEG may assist in this sometimes difficult determination. We report on routine screening EEGs in children hospitalized over an 18-month period for behavior problems. Eighty-six children were admitted for conduct disorder, attention deficit hyperactivity disorder, or both. Seventy-eight tracings (91%) were normal or showed normal variant patterns. Eight records (9%) were abnormal, showing background slowing or paroxysmal discharges not associated with behavioral manifestations. None of these neurologically normal, nonretarded patients had epilepsy or other known cerebral disorder. This suggests that routine EEG screening may be of limited value in childhood behavior problems without clinical evidence of neurologic disorder. PMID- 8420695 TI - Latency of P3 in semantic categorization of Chinese characters: preliminary report. AB - ERPs to target stimulation with tone and semantic categorization of Chinese characters were studied in 100 healthy subjects. Age ranged from 40-50 years old (80 subjects) and 50-68 years old (20 subjects). Two hundred Chinese characters were divided into four groups in accordance with the meaning of characters. The probability of each group was 25%. Each group could be used as target stimulus. The latencies of the exogenous components on these two different tasks were the same. The endogenous components on the semantic categorization of Chinese characters were longer than with tone stimulation. In the 40-50 years age group the mean latencies of positive waves at Fz, Cz, C3, and Pz were 546.1034 msec, 547.8621 msec, 548.0345 msec and 547.3103 msec, respectively; latencies in the 50 68 years group were 542.9412 msec, 545.2941 msec, 543.8824 msec and 553.0588 msec respectively. A positive wave, which we called P550 and divided into P550a and P550b, could be recognized in 27.5% of them. PMID- 8420696 TI - Migraine-triggered epilepsy. AB - Eight young adult patients are reported, referred because of generalized tonic clonic seizures with unsatisfactory treatment response. All of them had migraine. The major convulsions were preceded by a typical visual prodroma in 6 out of 8 patients, and a full-blown migraine attack followed the convulsion in all patients. Clinical findings were unremarkable, the EEG was mostly normal or slightly abnormal without typical paroxysmal findings, and CT scan and MRI were normal. All patients also had independent migraine attacks without convulsions; all of them had a positive family history of migraine. The response to antimigrainous and/or antiepileptic medication was sluggish or disappointing in most cases. A correction of the patient's life style proved to be the most effective approach. PMID- 8420697 TI - Episodic coma due to acute basilar artery migraine: correlation of EEG and brainstem auditory evoked potential patterns. AB - Our patient presented with three episodes of deep coma in 5 weeks, followed by a complete recovery. The neuroradiological tests and spinal fluid analysis excluded structural lesions, including subarachnoid hemorrhage, from the diagnosis. A nonconvulsive status was excluded by 24-hour EEG monitoring. The EEGs and brainstem auditory evoked responses were abnormal during ictus, but they reversed to normal with the clinical recovery. Timely neurophysiological tests helped in the diagnosis of basilar artery migraine. PMID- 8420698 TI - Midline theta rhythm revisited. AB - The midline theta rhythm consists of runs of theta waves occurring in the midline, especially in the central and parietal vertex regions. Few reports regarding this uncommon EEG finding have been published. It has been given various names, e.g. theta discharges in the middle-line, theta spindles, etc. Out of the 4,236 patients who were administered EEG examinations during an investigation period of 9 months, the midline theta rhythm, with a frequency in the range of 4-7 Hz, voltage over 50 microV, duration over 3 seconds, and most prominent in the central and parietal vertex areas, was recorded in 35 (0.83%). The age distribution of these patients peaked in two groups, adults aged 21-40 years and elderly patients aged 61-77 years. The waveform of the theta rhythm observed in adults 40 years old and under was often sinusoidal and regular. In this group, the most common clinical disorder was epilepsy. Often, the appearance of the theta rhythm observed in elderly patients was triangular and irregular, and sometimes sharp. In this group, the common clinical symptoms were headaches and dizziness, characterized by diagnoses of organic brain diseases such as cerebrovascular disorders. This study re-examines the clinical significance of the midline theta rhythm, taking into consideration studies that have been published in the past. PMID- 8420699 TI - Aging, sun damage, and sunscreens. AB - Intrinsic aging deleteriously changes the structure and function of human skin. Acute and chronic sun exposure may accelerate the aging processes in skin. Patients should avoid sun during the peak hours of sunshine, wear protective clothing, and use sunscreens while outdoors. Acceptable medical and surgical treatments may be offered to patients who manifest the cutaneous consequences of aging and sun exposure. PMID- 8420700 TI - Dysplastic and congenital nevi. AB - The dysplastic nevus, both the sporadic and familial type, has a significant association with melanoma of the skin. The congenital nevus, which ranges in size from less than 1 cm to a giant hairy nevus, are treated primarily for aesthetic reasons and, in the giant variety, are also treated as a prophylaxis against the possibility of malignant change. PMID- 8420701 TI - Nonepidermal and appendageal skin tumors. AB - This article reviews the essential clinical and pathologic features of a number of tumors of the dermis and epidermal appendages to help improve the clinician's skill at formulating preoperative differential diagnoses, assessing the need for treatment, and determining appropriate follow-up. In addition, it attempts to alert the practitioner to a number of heritable and systemic conditions that may be signalled by the presence of cutaneous tumors. PMID- 8420702 TI - Cryosurgery. AB - This article has discussed the use of cryosurgery as a method for treating benign, premalignant, and malignant skin lesions. Treatment of acne has also been discussed. Correct freeze times for obtaining good cosmetic results and a high cure rate have been stressed. Methods for obtaining high cure rates in treating skin cancer have been given. Modern day cryosurgery is an efficient, effective modality yielding good cosmetic results and high cure rates. PMID- 8420703 TI - Mohs micrographic surgery. AB - This exacting surgical technique with histologic examination of frozen sections is only applicable to tumors that spread locally by contiguous cell extension into local tissue. The narrow margins obtained with this technique may preserve function and cosmesis when basal or squamous cell carcinomas are resected and the defect is reconstructed. PMID- 8420704 TI - Reconstruction after Mohs cancer excision. AB - At our institution, 363 skin defects following Mohs excision for carcinoma were repaired in a two-year period. The majority of the patients were women (62%). Most repairs were to the nose (42%), and almost all followed basal cell carcinoma excision (91% of tumor types). Flaps were preferable to skin grafts for facial repair, with forehead and nasolabial flaps particularly useful for the nose. Injection of Kenalog (triamcinolone acetonide, 5-20 mg/mL) speeds the maturation of scars and flaps. PMID- 8420705 TI - Injectable soft tissue substitutes. AB - Historical and modern advances in the development of an injectable soft tissue substitute are reviewed. Nonbiologic alloplastic and biologic injectables are described. The authors' experiences, as well as those of others, employing presently available materials in terms of specific indications and special techniques are delineated. The search for a safe, effective, easy-to-use, and long-lasting soft tissue substitute continues. PMID- 8420706 TI - Considerations for cosmetic surgery in the black population. AB - The availability of cosmetic surgical procedures to the general public, especially blacks, has become increasingly widespread. Aesthetic surgeons should not be deterred from performing these procedures in blacks. The myth that all black patients develop keloids or dyspigmentation after surgery should be dispelled; however, in those patients with a history of keloid formation or hypertrophic scarring, elective cosmetic procedures should be either withheld or performed with extreme caution. In general, the cosmetic surgeon can proceed with surgery in blacks and attain good aesthetic results if the procedures are slightly modified when indicated to minimize pigmentary changes and scarring and to maintain ethnic congruency. PMID- 8420707 TI - Acne and related disorders. AB - Acne vulgaris is the clinical expression of inflammation of the pilosebaceous unit. Factors known to predispose to the development of acne include increased sebum, which is acted on by Propionobacterium acnes to generate inflammatory substances, and retention hyperkeratosis, which causes obstruction of the sebaceous follicle. Therapeutic modalities for acne include topical and systemic antibiotics, comedolytic agents (such as benzoyl peroxide and topical retinoids) and systemic retinoids. Acne scars may be treated surgically using procedures such as dermabrasion and dermal injections of bovine collagen or simple scar excision, scar punch elevation, or punch grafting. PMID- 8420708 TI - Vascular lesions. AB - Most cutaneous vascular abnormalities can be successfully, and in fact preferentially, treated with lasers. The field is evolving rapidly. It is important that the appropriate laser be used. In addition, many of the early references in this field are quite out of date. We are treating lesions at a much earlier age and our expectations are much higher. We expect the result to be virtually normal-looking skin with complete resolution of the problem. Capillary vascular malformations (port wine stain) are the most frequent indication for laser treatment in both children and adults. With the advent of the flashlamp pumped pulsed dye laser, these lesions can be treated in infancy and early childhood. This represents a significant breakthrough, because unlike with previous lasers, scarring is a rare side effect of treatment with yellow light lasers. The psychologic trauma of growing up with a facial deformity can be minimized with early treatment. It is hoped that the hypertrophy and permanent deformity associated with these lesions can be mitigated. The various lasers currently available to the plastic surgeon provide treatment options not previously available. Lasers are expected to play an increasing role in cutaneous abnormalities in the field of plastic surgery. PMID- 8420709 TI - Pigmentary changes in the skin. An introduction for surgeons. AB - Alterations in pigmentation of the skin are commonly encountered as a patient's primary reason for seeking medical care. They may be discovered as incidental findings on examination of the skin. The physician should be able to recognize and treat some of the common pigmentary disorders and to make referrals when appropriate. It is important to be aware of associations of pigmentary changes with systemic diseases. PMID- 8420710 TI - Infectious disorders of the skin. AB - Cutaneous infections are common occurrences in clinical practice. Plastic surgeons need to recognize the varied presentations of cutaneous infections because they may mimic cutaneous malignancies or be a harbinger of underlying disease. We have reviewed the diagnosis and management of some of the more common bacterial, viral, and fungal infections of the skin. PMID- 8420711 TI - Connective tissue diseases. AB - The classic connective tissue diseases (lupus erythematosus, dermatomyositis, and scleroderma) have been described with respect to their pathogenesis, clinical picture, laboratory diagnosis, and management. Because the aim of this volume is to provide an interface between plastic surgery and dermatology, the cutaneous manifestations, both shared and pathognomonic for each of these diseases, have been emphasized. An attempt has been made to point out opportunities for surgical management of some of the problems associated with these diseases. PMID- 8420712 TI - Treatment of photoaging. Facial chemical peeling (phenol and trichloroacetic acid) and dermabrasion. AB - A complete armamentarium using phenol, trichloroacetic acid, and dermabrasion allows the physician to successfully treat a variety of difficult photoaged skin problems in a consistent fashion. These three techniques have their specific indications, and patient selection is the key to a successful outcome. Proper attention to technical detail will allow the physician to fine-tune technique to meet the individual's needs. It is important to realize that phenol, trichloroacetic acid, and dermabrasion are not exclusive of each other, but are additive in their value. As one becomes well versed in these differing treatment modalities, one can tailor these techniques to obtain consistent results according to the needs and desires of the patient. PMID- 8420713 TI - Nonexcisional treatment of benign and premalignant cutaneous lesions. AB - The correct diagnosis of benign and premalignant cutaneous lesions requires appropriate biopsy techniques that reflect an understanding of the specific cutaneous pathology. Violation of the basement membrane zone generally results in scarring. Knowledge of the healing processes of the epidermis, dermis, and subcutis permit the least destructive diagnostic and treatment techniques, resulting in the best functional and cosmetic results. Excessive scarring or procedures inappropriate to the lesion under consideration should be avoided. PMID- 8420714 TI - Injuries of the knee. PMID- 8420715 TI - Acute brain swelling after out-of-hospital cardiac arrest: pathogenesis and outcome. AB - OBJECTIVES: First, to examine factors that may be related to brain swelling, which was identified by the absence or compression of the lateral and third ventricles and perimesencephalic cisterns on brain computed tomography (CT) scans in the early postresuscitation period in patients who suffered an out-of-hospital cardiac arrest. Second, to characterize the neurologic outcome in those patients in whom cardiac arrest was followed by brain swelling. DESIGN: Prospective and retrospective analyses. SETTINGS: General ICU, tertiary care hospital. PATIENTS: Fifty-three patients (35 male, 18 female) who had an out-of-hospital cardiac arrest and who also had a brain CT examination on the third day after resuscitation. The 53 patients were divided into two groups: group A (25 patients) experienced brain swelling on postresuscitation day 3; group B (28 patients) did not experience noticeable brain swelling. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: There was a significant difference between the two groups in the etiology of the cardiac arrest. Twenty-three of 25 patients in group A had cardiac arrest due to respiratory distress, whereas this finding was true in only five patients in group B. In laboratory data, arterial pH was significantly lower in group A than in group B (6.93 vs. 7.09), as was base deficit (-21.0 mmol/L in group A vs. -13.7 mmol/L in group B). Neurologic outcome was evaluated 1 wk after resuscitation. There were significantly more patients in group A who were not awake and who were diagnosed as brain dead. CONCLUSIONS: The cause of brain swelling may be related to the development of the metabolic acidosis (possibly lactic acidosis) due to hypoxia before the resuscitation period. Brain swelling may be one of the indicators that predicts a poor neurologic outcome in the patients who suffer an out-of-hospital cardiac arrest. PMID- 8420716 TI - Pharmacokinetics of exogenous epinephrine in critically ill children. AB - OBJECTIVES: This study was designed to determine the steady-state plasma concentrations and clearance rates of epinephrine in critically ill children, to examine if epinephrine pharmacokinetics conform to a linear model, and to compare epinephrine clearance rates with clearance rates of dopamine and dobutamine. DESIGN: This study was prospective, without intervention or control groups. SETTING: The pediatric ICUs of two tertiary care teaching hospitals. PATIENTS: All patients who were hemodynamically stable while requiring continuous epinephrine infusions were eligible for the study. INTERVENTIONS: Blood samples were taken at steady state and analyzed for epinephrine concentrations, as well as dopamine and dobutamine concentrations, if present, by high-pressure liquid chromatography with electrochemical detection. RESULTS: Plasma epinephrine concentrations during steady-state infusions of 0.03 to 0.2 micrograms/kg/min ranged from 670 to 9430 pg/mL (3660 to 51,490 pmol/L), with a mean of 4360 +/- 3090 pg/mL (23,810 +/- 16,870 pmol/L) and were linearly related to dose. Epinephrine clearance rates ranged from 15.6 to 79.2 mL/kg/min (mean 29.3 +/- 16.1) and were not dependent on steady-state plasma concentrations. Epinephrine clearance rate was in the same range as the clearance rates of dopamine (34.1 +/- 16.6) and dobutamine (35.9 +/- 27.8) and was linearly related to them (p < .005). CONCLUSIONS: Epinephrine infusions produce pharmacologic plasma concentrations of epinephrine in critically ill children. The plasma concentration of epinephrine correlates with the infusion rate, suggesting linear pharmacokinetics. Epinephrine clearance rates in critically ill children appear to be lower than the reported clearance rates in healthy adults. The clearance rates of two other inotropic catecholamines, dopamine and dobutamine, are significantly correlated with the clearance rate of epinephrine. PMID- 8420717 TI - Synchronous mechanical ventilation of the neonate with respiratory disease. AB - OBJECTIVES: To assess the importance of synchronization of mechanical ventilation with spontaneous respiratory efforts in mechanically ventilated neonates. The actions of this synchronization on ventilation, oxygenation, and BP variation were assessed. DESIGN: Prospective evaluation using within-subject comparison of asynchronous and synchronous states. SETTING: Neonatal ICU in a large, university affiliated hospital. PATIENTS: Fourteen neonates requiring mechanical ventilation who were initially asynchronous with the ventilator. INTERVENTION: The ventilator settings were adjusted using the patients' own inspiratory and expiratory timing to create synchronous interaction with the ventilator. MEASUREMENTS AND MAIN RESULTS: Synchrony was assessed using clinical observation combined with inspection of the air flow waveform and computerized analysis of the air flow signal to assess cycle-to-cycle reproducibility, so-called autocorrelation. Synchronous ventilation significantly improved tidal volume (p < .05), minute volume (p < .001), and all indices of the variability of arterial BP (p < .001). Mean airway pressure did not change significantly. No infant developed an airleak syndrome or intraventricular hemorrhage, which have previously been associated with asynchronous ventilation and an unstable BP, respectively. CONCLUSION: Synchronous ventilation can be readily applied to most ventilated neonates. It improves ventilation, and results in a marked reduction in BP variation, which may have implications for reducing the risk of intraventricular hemorrhage. PMID- 8420718 TI - Predicting mortality risk for infants weighing 501 to 1500 grams at birth: a National Institutes of Health Neonatal Research Network report. AB - OBJECTIVES: To develop and evaluate a model that predicts mortality risk based on admission data for infants weighing 501 to 1500 grams at birth, and to use the model to identify neonatal ICUs where the observed mortality rate differs significantly from the predicted rate. DESIGN: Validation cohort study. SETTING: University-based, tertiary care neonatal ICUs. PATIENTS: Sample of 3,603 infants with birth weights of 501 to 1500 grams who were born at seven National Institute of Child Health and Human Development (NICHHD) Neonatal Research Network Centers, over a 2-yr period of time. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Based on logistic regression analysis, admission factors associated with mortality risk for inborn infants were: decreasing birth weight, appropriate size for gestational age, male gender, non-black race, and 1-min Apgar score of < or = 3. The mortality prediction model based on these factors had a sensitivity of 0.50, a specificity of 0.92, a correct classification rate of 0.82, and an area under the receiver operating characteristic curve of 0.82 when applied to a validation sample. Goodness-of-fit testing showed that there was a marginal degree of fit between the observations and model predictions (chi 2 = 15.4, p = .06). The observed mortality rate for 3,603 infants at the seven centers was 24.7%, ranging from 21.8% to 27.7% at individual centers. There were no statistically significant differences between observed and predicted mortality rates at any of the centers. One center had an observed mortality rate that was 2.8% lower than predicted by the model (95% confidence interval -6.0% to 0.5%), and another center had an observed rate that was 3% higher than expected (95% confidence interval -0.3% to 6.2%). CONCLUSIONS: Mortality risk for infants weighing 501 to 1500 grams can be predicted based on admission factors. However, until more accurate predictive models are developed and validated and the relationships between care practices and outcomes are better understood, such models should not be relied on for evaluating the quality of care provided in different neonatal ICUs. PMID- 8420719 TI - Effects of dense, high-volume, artificial surfactant aerosol on a heated exhalation filter system. AB - OBJECTIVE: To evaluate a supplemental heated filter system during mechanical ventilation with continuous nebulization of an artificial surfactant by a new, high-volume nebulizer. DESIGN AND METHODS: A new nebulizer system, containing artificial surfactant, provided half of a 20-L minute ventilation and the remainder of this minute ventilation was provided by a commonly used mechanical ventilator. Ventilation sources were joined in the inspiratory limb of the breathing circuit, which was connected to a test lung system. A supplemental filter system was placed upstream of the ventilator's heated filter in the expiratory limb of the circuit. Circuit pressures at the inlet of the supplemental filter (P1), between the filters (P2), and after the ventilator expiratory filter (P3) were monitored and recorded. Nebulizer canisters containing artificial surfactant were replaced every 4 hrs. The performance of four supplemental filters in continuous use was examined. Another four filters were each used over 4 hrs, steam autoclaved, and reused. SETTINGS: The ventilator was set at a rate of 20 breaths/min, with a tidal volume of 0.5 L, a flow rate of 40 L/min, and positive end-expiratory pressure of 10 cm H2O. The nebulizer provided an equal volume and flow rate so that the delivered tidal volume was 1.0 L with a flow rate of 80 L/min. RESULTS: Ventilator failure and/or excessive airway pressure caused by increased filter resistance occurred at a mean of 7.3 +/- 1.3 (SD) hrs of continuous ventilation. Mean P1-peak increased from 67.5 +/- 8.2 to 94.0 +/- 10.7 cm H2O (p < .001) and P1-baseline increased from 9.3 +/- 1.0 to 53.5 +/- 17.1 cm H2O (p = .014). Filters that were autoclaved after 4 hrs of ventilation and reused lasted a total of 7.0 +/- 1.3 hrs. Mean P1-peak increased from 68.9 +/- 4.9 to 84.8 +/- 19.1 cm H2O and P1-baseline increased from 9.5 +/- 1.7 to 30.8 +/- 14.2 cm H2O (p < .05). CONCLUSIONS: The supplemental filter system was able to protect the ventilatory exhalation sensors for approximately 7 hrs at a minute ventilation of 20 L/min. Steam sterilization did not extend the supplemental filter life. PMID- 8420720 TI - Mechanisms of ventilator-induced lung injury. AB - OBJECTIVES: To describe the physiologic mechanisms of ventilator-induced lung injury and to define the major ventilator and host-dependent risk factors that contribute to such injury. DATA SOURCE: Basic science and clinical studies related to ventilator-induced barotrauma and lung pathophysiology. STUDY SELECTION: Emphasis on controlled, experimental studies and clinical studies related to specific mechanisms. DATA EXTRACTION: Preference given to studies with quantitative end-points to assess damage and causal relationships. DATA SYNTHESIS: Related studies are integrated to obtain basic mechanisms of damage where possible. CONCLUSIONS: Ventilation with high tidal volumes can increase vascular filtration pressures; produce stress fractures of capillary endothelium, epithelium, and basement membrane; and cause lung rupture. Mechanical damage leads to leakage of fluid, protein, and blood into tissue and air spaces or leakage of air into tissue spaces. This process is followed by an inflammatory response and possibly a reduced defense against infection. Predisposing factors for lung injury are high peak inspiratory volumes and pressures, a high mean airway pressure, structural immaturity of lung and chest wall, surfactant insufficiency or inactivation, and preexisting lung disease. Damage can be minimized by preventing overdistention of functional lung units during therapeutic ventilation. PMID- 8420721 TI - A survey of charting in critical care units. AB - OBJECTIVES: To determine what data are currently being collected at the critical care bedside, the role of flow sheets in storing these data, and what other forms and locations are used to store critical care data. DESIGN: We undertook a descriptive survey in a nonrandom sample of acute care hospitals with designated critical care units. Flow sheets were reviewed, and interviews with critical care nurses and physicians were conducted according to a predefined questionnaire. Finally, compliance in the surveyed hospitals with previous recommendations and existing guidelines on data collection and storage was then determined. SETTING: Fifteen "teaching" and 19 "nonteaching" acute care hospitals in the Province of Ontario. RESULTS: A lack of standardization of flow-sheet design and of the specific data included on these flow sheets was identified. Abbreviations were inconsistent. Little collaboration between physician and nonphysician members of the critical care team was identified in flow-sheet design. Variance between units in their collection of physiologic data was remarkable. Finally, few units surveyed used quantifiable illness severity scoring to describe their unit population. CONCLUSIONS: The lack of standardization regarding key information that should be collected and archived in critical care units identifies important risk management and quality assurance issues. There is a need for agreement on what information should be collected and maintained at the bedside in order to provide quality patient care. PMID- 8420722 TI - Rhabdomyolysis associated with status asthmaticus. PMID- 8420723 TI - Indomethacin and apnea of prematurity. PMID- 8420724 TI - Treatment of gram-negative septic shock with an immunoglobulin preparation: a prospective, randomized clinical trial. PMID- 8420725 TI - Relationship between cardiac index, mixed venous blood oxygen saturation and regional venous blood oxygen saturation. PMID- 8420726 TI - Continuous intravenous cimetidine decreases stress-related upper gastrointestinal hemorrhage without promoting pneumonia. AB - OBJECTIVES: To determine whether a continuous i.v. infusion of cimetidine, a histamine-2 (H2) receptor antagonist, is needed to prevent upper gastrointestinal (GI) hemorrhage when compared with placebo and if that usage is associated with an increased risk of nosocomial pneumonia. Due to the importance of this latter issue, data were collected to examine the occurrence rate of nosocomial pneumonia under the conditions of this study. DESIGN: A multicenter, double-blind, placebo controlled study. INTERVENTIONS: Patients were randomized to receive cimetidine (n = 65) as an iv infusion of 50 to 100 mg/hr or placebo (n = 66). SETTING: Intensive care units in 20 institutions. PATIENTS: Critically ill patients (n = 131), all of whom had at least one acute stress condition that previously had been associated with the development of upper GI hemorrhage. MEASUREMENTS AND MAIN RESULTS: Samples of gastric fluid from nasogastric aspirates were collected every 2 hrs for measurement of pH and were examined for the presence of blood. Upper GI hemorrhage was defined as bright red blood or persistent (continuing for > 8 hrs) "coffee ground material" in the nasogastric aspirate. Baseline chest radiographs were performed and sputum specimens were collected from all patients, and those patients without clear signs of pneumonia (positive chest radiograph, positive cough, fever) at baseline were followed prospectively for the development of pneumonia while receiving the study medication. Cimetidine-infused patients experienced significantly (p = .009) less upper GI hemorrhage than placebo-infused patients: nine (14%) of 65 cimetidine vs. 22 (33%) of 66 placebo patients. Cimetidine patients demonstrated significantly (p = .0001) higher mean intragastric pH (5.7 vs. 3.9), and had intragastric pH values at > 4.0 for a significantly (p = .0001) higher mean percentage of time (82% vs. 41%) than placebo patients. Differences in pH variables were not found between patients who had upper GI hemorrhage and those patients who did not, although there was no patient in the cimetidine group who bled with a pH < 3.5 compared with 11 such patients in the placebo group. Also, the upper GI hemorrhage rate in patients with one risk factor (23%) was similar to that rate in patients with two or more risk factors (25%). Of the 56 cimetidine-infused patients and 61 placebo-infused patients who did not have pneumonia at baseline, no cimetidine-infused patient developed pneumonia while four (7%) placebo-infused patients developed pneumonia. CONCLUSIONS: The continuous i.v. infusion of cimetidine was highly effective in controlling intragastric pH and in preventing stress-related upper GI hemorrhage in critically ill patients without increasing their risk of developing nosocomial pneumonia. While the number of risk factors and intragastric pH may have pathogenic importance in the development of upper GI hemorrhage, neither the risk factors nor the intragastric pH was predictive. Therefore, short-term administration of continuously infused cimetidine offers benefits in patients who have sustained major surgery, trauma, burns, hypotension, sepsis, or single organ failure. PMID- 8420727 TI - Can we predict mortality for low birth weight infants? PMID- 8420728 TI - Stress ulcer prophylaxis: in whom? With what? PMID- 8420729 TI - Iatrogenic complications in adult intensive care units: a prospective two-center study. AB - OBJECTIVES: a) To evaluate the frequency, types, severity, and morbidity of iatrogenic complications; b) determine associated factors that favor iatrogenic complications; and c) suggest new or more efficient protective measures that may be taken to improve patient safety. DESIGN: Prospective, observational study. SETTING: Two ICUs in France. PATIENTS AND METHODS: The study included 382 patients (age > or = 15 yrs; 400 consecutive admissions). Patients were monitored by two physicians in each ICU to assess all iatrogenic complications occurring during their ICU stay, with the exception of adverse effects of drugs. An iatrogenic complication was defined as an adverse event that was independent of the patient's underlying disease. RESULTS: We observed 316 iatrogenic complications in 124 (31%) of the 400 admissions. Of these iatrogenic complications, 107 (in 53 [13%] of the 400 admissions) complications were major, three leading to death. Severe hypotension, respiratory distress, pneumothorax, and cardiac arrest represented 78% of the major iatrogenic complications. Fifty nine percent of the major iatrogenic complications had clearly identified associated factors. Human errors accounted for 67% of these factors. Patients > 65 yrs (adjusted odds ratio = 2.6, 95% confidence interval: 1.4 to 4.9) and those patients admitted with two or more organ failures (adjusted odds ratio = 4.8, 95% confidence interval: 2.5 to 9.2) were more likely to develop major iatrogenic complications. High or excessive nursing workload also led to an increased risk of major iatrogenic complications. Persistent morbidity, secondary to iatrogenic complications at the time of discharge, was present in five survivors. The risk of ICU death was about two-fold higher for the patients with major iatrogenic complications than in the remaining patients after adjusting for the Organ System Failure Score and the prognosis of the disease (relative risk = 1.92, 95% confidence interval: 1.28 to 2.56). CONCLUSIONS: Major iatrogenic complications were frequent, associated with increased morbidity and mortality rates, related to high or excessive nursing workload, and were often secondary to human errors. To improve patient safety in our ICUs, preventive measures should be targeted primarily on the elderly and the most severely ill patients. Special attention should be given to improving the organization of workload and training, and promoting wider use of noninvasive monitoring. PMID- 8420731 TI - Relationship between supranormal circulatory values, time delays, and outcome in severely traumatized patients. AB - OBJECTIVE: To describe the temporal patterns of hemodynamics and oxygen transport in survivors and nonsurvivors of severe trauma in relation to time delays, mortality, and morbidity. DESIGN: Prospective, empiric analysis. SETTING: University-run, inner city county hospital with a Level I trauma center. PATIENTS: A series of 90 consecutively monitored, severely ill trauma patients. METHODS: We followed 90 patients from admission through their hospital course, and divided the study group into patients with estimated blood loss of < or = 3000 mL and those patients with an estimated blood loss of < 3000 mL. For each patient, vital signs were recorded in the Emergency Department, operating room, recovery room, and surgical ICU. Hemodynamic and oxygen transport variables were measured at least every 12 hrs for 96 hrs postadmission. Final outcome and complications were recorded. RESULTS: In the first 24 hrs, the values of 60 survivors were significantly higher than the values of 30 nonsurvivors for mean cardiac index (4.52 +/- 1.45 vs. 3.80 +/- 1.20 L/min/m2; p < .05), oxygen delivery (670 +/- 230 vs. 540 +/- 200 mL/min/m2; p < .01), and oxygen consumption (166 +/- 48 vs. 134 +/- 47 mL/min/m2; p < .01). Thirteen (50%) of 26 patients who never attained mean survivors' values (defined as the mean survivors' values listed above) died. Also, 12 (57%) of 21 patients who took > 24 hrs to attain these values died. In contrast, only five (12%) of 43 patients who reached mean survivors' values in < or = 24 hrs died. Thirty-five of 90 patients lost < 3000 mL of blood; 17 of these 35 patients failed to reach survivors' values within 24 hrs, and 12 (71%) patients died. However, of 18 patients with an estimated blood loss of > 3000 mL, who reached survivors' values in < or = 24 hrs, only two (12%) died. The patients reaching survivors' values in < or = 24 hrs, > 24 hrs, or not at all had similar Injury Severity Scores (28 +/- 13, 26 +/- 13, and 26 +/- 12, respectively) and Trauma Scores (12 +/- 3, 13 +/- 3, and 12 +/- 3, respectively). Only six (12%) of 43 patients reaching survivors' values in < or = 24 hrs developed adult respiratory distress syndrome (ARDS), while 27 (57%) of 47 patients showed these values in > 24 hrs or never developed ARDS. CONCLUSIONS: Reaching supranormal circulatory values, especially within 24 hrs of injury, may improve survival and reduce the frequency of shock-related organ failure in severely traumatized patients. PMID- 8420730 TI - Do-not-resuscitate orders in a children's hospital. AB - OBJECTIVES: a) To quantify the use of do-not-resuscitate orders in a tertiary care children's hospital; and b) to characterize the circumstances in which such orders are written. DESIGN: Retrospective chart review. SETTING: University teaching hospital. PATIENTS: All inpatients who died in an urban children's hospital over a 1-yr period of time. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The hospital records of 54 of 69 patients who died were reviewed. Eighty two percent of patient deaths occurred in the ICU; 13% of patient deaths occurred in the operating room, and 5% occurred in a medical ward. Other findings included the following: 25 (46%) of 54 patients died after attempted cardiopulmonary resuscitation; 13 (24%) patients were brain dead; and 16 (30%) died with a do-not resuscitate order in effect. Age was associated with resuscitation status: do-not resuscitate orders were written for five (21%) of 22 infants (< 1 yr of age); seven (50%) of 14 children (1 to 11 yrs of age); and four (80%) of five adolescents who died (p < .002). Fifteen of 16 do-not-resuscitate orders were written for patients who were in the ICU, where they remained until death. Findings in patients when the do-not-resuscitate orders were written were as follows: 15 ICU patients were receiving mechanical ventilation; 14 (95%) of 15 were receiving inotropic agents; 12 (80%) of 15 patients were receiving narcotic analgesics; and one (6%) patient was being dialyzed. At least one therapeutic modality was withdrawn in 7 (44%) of 16 patients. Do-not-resuscitate orders followed documented conferences with physicians and family members in 13 (81%) of 16 cases. These discussions were initiated by physicians in 12 (92%) of 13 cases. CONCLUSIONS: Do-not-resuscitate orders in pediatric patients are written more often in older than younger hospitalized children who die. Most do-not resuscitate orders are written for patients who are receiving aggressive medical therapy in the ICU. PMID- 8420732 TI - Relationship between oxygen consumption and oxygen delivery during anesthesia in high-risk surgical patients. AB - OBJECTIVE: To identify critical oxygen delivery (DO2) and oxygen extraction ratio in high-risk surgical patients studied preoperatively and intraoperatively. DESIGN: Prospective study. Consecutive series of intraoperatively monitored patients. SETTING: The surgical ICU in a tertiary care center. PATIENTS: High risk surgical patients undergoing major noncardiac surgery. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Twenty high-risk patients were studied during the preoperative and intraoperative periods. All patients were monitored with triple lumen thermodilution catheters. Hemodynamic profiles consisted of determinations of intravascular pressures, cardiac output, and arterial and venous pulmonary gases. Oxygen transport variables were calculated by standard formulas. Multiple determinations were carried out during the perioperative period in each patient. The critical levels of DO2, determined by a polynomial regression method, were 375 and 390 mL/min/m2 in the preoperative and intraoperative periods, respectively. Oxygen extraction reached at the critical level of DO2 was significantly (p < .01) lower during the intraoperative period (31 +/- 4.5% vs. 18 +/- 2.3%). Critical mixed venous oxygen tension was significantly (p < .01) higher in the intraoperative period (36 +/- 5 vs. 46 +/- 4 torr [4.8 +/- 0.7 vs. 6.1 +/- 0.5 kPa]). CONCLUSIONS: Our data show that the intraoperative period might be associated with a reduction in tissue ability to extract oxygen. If this reduction in oxygen extraction is proportionately higher than the reduction in metabolic oxygen demand produced by anesthetic agents and hypothermia, then the critical value for DO2 may be similar to, or higher than, that value in the preoperative period. Thus, the intraoperative period represents, for this patient population, a high-risk condition for the development of a tissue oxygenation debt in the presence of a limitation in DO2. Cautious dosing of inhaled agents, maintenance of normothermia, and early optimization of the oxygen delivery/oxygen consumption relationship seem to constitute reasonable measures in the intraoperative handling of these patients in order to reduce perioperative morbidity and mortality. PMID- 8420733 TI - Epinephrine as an inotropic agent in septic shock: a dose-profile analysis. AB - OBJECTIVE: To characterize the acute actions and physiologic dose profile of epinephrine, as a single inotrope, in patients with septic shock. DESIGN: Prospective clinical study. The relationship between epinephrine dose and cardiovascular variables was analyzed using repeated-measures analysis of variance. SETTING: ICU in a university teaching hospital. PATIENTS: Eighteen patients with septic shock, mean age 64 +/- 8 (SD) yrs, and with a mean admission Acute Physiology and Chronic Health Evaluation (APACHE II) score of 23 (range 14 to 35). INTERVENTIONS: Initial volume loading and the measurement of a baseline hemodynamic profile were followed by the administration of an epinephrine infusion at 3 microgram/min with subsequent increments of 3 micrograms/min and the determination of a hemodynamic profile after each dose increment. Therapy was titrated to clinical goals of perfusion and restoration of premorbid systolic arterial BP. MEASUREMENTS AND MAIN RESULTS: After volume loading, mean hemodynamic indices were as follows: mean arterial pressure (MAP) 62 +/- 7 mm Hg; cardiac index 3.8 +/- 1.1 L/min/m2; left ventricular stroke work index 25 +/- 11 g.m/m2; oxygen delivery (Do2) index 460 +/- 168 mL/min/m2; and oxygen consumption (VO2) index 165 +/- 64 mL/min/m2. In the dose range of 3 to 18 microgram/min, epinephrine produced linear increases in average heart rate, MAP, cardiac index, left ventricular stroke work index, stroke volume index, VO2, and DO2. No effect was noted on pulmonary artery occlusion pressure (PAOP), mean pulmonary arterial pressure, or systemic vascular resistance index. CONCLUSIONS: Epinephrine increases DO2 in septic shock by increasing cardiac index without an effect on systemic vascular resistance index or PAOP. PMID- 8420734 TI - Respiratory mechanics and bronchodilator responsiveness in patients with the adult respiratory distress syndrome. AB - OBJECTIVE: To study the effects of salbutamol (a selective beta 2-adrenergic receptor agonist) on respiratory mechanics in patients with the adult respiratory distress syndrome (ARDS). DESIGN: Prospective study. SETTING: ICU in a university hospital. PATIENTS: Seven mechanically ventilated, paralyzed ARDS patients. MAIN OUTCOME MEASUREMENTS: Measurements of respiratory system compliance, maximum, and minimum inspiratory resistance (by the end-inspiratory occlusion method during constant flow inflation) were performed at 0, 5, 10 cm H2O positive end expiratory pressure, both before and at least 30 mins after the start of a continuous iv infusion of salbutamol (15 micrograms/min). Minimum inspiratory resistance represents the ohmic air flow resistance, while maximum inspiratory resistance includes minimum inspiratory resistance plus the effective additional resistance due to stress adaptation and to time constant inhomogeneities. Air flow was measured at the airway connector and tracheal pressure near the central end of the artificial airway. RESULTS: Maximum inspiratory resistance, minimum inspiratory resistance, and additional resistance were higher than the values reported for normal anesthetized subjects. On average, salbutamol caused a decrease in maximum and minimum inspiratory resistances (from 6.48 +/- 2.56 to 4.67 +/- 1.74 and from 4.06 +/- 2.12 to 2.07 +/- 0.95 cm H2O/L/sec, respectively). Positive end-expiratory pressure increased additional resistance, whereas it decreased minimum inspiratory resistance. No interaction was found between positive end-expiratory pressure and salbutamol. Respiratory system compliance was not significantly affected by salbutamol nor by positive end expiratory pressure. CONCLUSIONS: In ARDS patients, salbutamol decreases the abnormally high airway resistance, by reducing minimum resistance, but has no effect on the effective additional resistance. PMID- 8420735 TI - Iatrogenic complications in the intensive care unit. PMID- 8420736 TI - Heparin in experimental hyperdynamic sepsis. AB - OBJECTIVE: To evaluate the hypothesis that heparin administration increases cardiac output and improves oxygenation in experimental hyperdynamic sepsis in sheep. DESIGN: Prospective trial. SETTING: Laboratory at a large university affiliated medical center. SUBJECTS: A total of 14 sheep weighing 28 to 44 kg. INTERVENTIONS: All 14 chronically instrumented sheep received a continuous infusion of Escherichia coli endotoxin (10 ng/kg/min) over 24 hrs. Seven sheep received a fixed bolus of beef lung heparin (5000 units) every 4 hrs intravenously. The other seven sheep served as controls. MEASUREMENTS AND MAIN RESULTS: The heparinized animals showed a triphasic cardiovascular response. Cardiac index increased (p < .05), and systemic vascular resistance index decreased (p < .05) at 2 hrs after the start of the endotoxin infusion (early phase, 0 to 2 hrs). Both variables returned to approximately baseline levels at 4 hrs (second period, 2 to 4 hrs). A hyperdynamic state (in terms of an increased cardiac index), a decreased systemic vascular resistance index, and a decreased mean arterial pressure (MAP) (p < .05 for all) was observed in the third phase (8 to 24 hrs). In the control group, cardiac index, systemic vascular resistance index, and MAP showed no changes in the first period, but a slight decrease in cardiac index and a slight increase in systemic vascular resistance index in the second period. The onset of the hyperdynamic state was less pronounced in the control group and cardiac index was lower (p < .05); likewise, systemic vascular resistance index was increased (p < .05) when compared with heparinized animals. Both groups developed pulmonary hypertension during the endotoxin infusion. The gas exchange in the heparin group was significantly impaired in the first and second periods, but returned to baseline levels in the hyperdynamic phase, whereas the oxygenation of the nonheparinized animals showed only minor changes in the first and second phases, but deteriorated significantly during the third phase of endotoxemia. CONCLUSIONS: In this experimental model of hyperdynamic sepsis, heparin significantly influenced the cardiopulmonary performance. Heparin preserved gas exchange and increased cardiac output but lowered systemic vascular resistance and MAP in the hyperdynamic state. PMID- 8420737 TI - Pediatric do-not-resuscitate orders. PMID- 8420738 TI - Alterations in feline tracheal permeability after mechanical ventilation. AB - OBJECTIVES: Previous investigations of ventilator-induced airway injury focused on histopathologic changes associated with various ventilators and strategies for their use. We hypothesized that mechanical ventilation is associated with alterations in tracheal epithelial permeability, and designed a study using an animal model to evaluate changes in tracheal epithelial permeability after administering different types of mechanical ventilation to test this hypothesis. DESIGN: Prospective, multiple-group, controlled trial. Five groups of animals were studied and compared. Eight animals were studied without intubation or mechanical ventilation. A total of 28 animals (seven in each group) were studied after conventional mechanical ventilation, high-frequency positive-pressure ventilation, high-frequency jet ventilation, or high-frequency flow interruption at respiratory rates of 20, 150, 400, and 900 breaths/min, respectively. Comparison of data for each group was done using the Kruskall-Wallis analysis of variance. Between-group comparisons were made using standard error of the mean comparisons. For airway pressures and other physiologic data, one-way analysis of variance was performed. Between-group comparisons were made using the Student Newman-Keuls' test. SETTING: Small animal physiology laboratory. SUBJECTS: Thirty six adult cats. INTERVENTIONS: Mechanically ventilated animals were treated for 8 hrs and then killed. Inspired oxygen concentration, BP, and mean airway pressures were comparable in mechanically ventilated animals. Spontaneously breathing control animals were killed without endotracheal intubation or exposure to mechanical ventilation. MEASUREMENTS AND MAIN RESULTS: Permeability values in isolated tracheal segments were calculated for 14C-sucrose, 3H-inulin, and fluorescein isothiocyanate-dextran-20. Tracheal epithelial permeability to all studied molecules increased after exposure to mechanical ventilators. These different mechanical ventilators increased epithelial permeability in a progressive manner that paralleled ventilatory frequency. The changes were greatest after ventilation at the highest frequency. These observed changes in tracheal permeability are consistent with previously observed alterations in tracheal histopathology after exposure to mechanical ventilation. CONCLUSIONS: Mechanical ventilation was associated with increases in tracheal permeability to large and small nonionic molecules. These changes occurred with all studied ventilators, used as they are clinically. Permeability changes paralleled ventilatory rate changes. PMID- 8420739 TI - Neurologic complications of critical medical illnesses. AB - OBJECTIVES: To identify the neurologic complications of critical medical illnesses, and to assess their effect on mortality rates and on medical ICU and hospital lengths of stay. DESIGN: Prospective clinical evaluation of all medical ICU admissions for 2 yrs. SETTING: A 14-bed, general medical intensive and coronary care unit in a large university hospital. PATIENTS: Patients (n = 1,850) admitted to the hospital, of whom 92 were admitted for primarily neurologic problems. Of the remaining 1,758 patients, 217 (12.3%) experienced a neurologic complication. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients developing a neurologic complication while in the medical ICU demonstrated an increased risk of inhospital mortality when compared with patients who did not suffer such problems (45.7% vs. 26.6%; p < .00001). Patients with neurologic complications experienced 2.5-fold longer medical ICU stay times (p < .001) and almost two-fold longer hospital stay times (p < .001). Metabolic encephalopathy, seizures, hypoxic-ischemic encephalopathy, and stroke were the most common complications. Sepsis was the most frequent cause of encephalopathy, and cerebrovascular lesions were the most common cause of seizures. Formal neurologic consultations were requested in only 36% of these patients. CONCLUSIONS: Neurologic complications are associated with increased mortality rates and longer medical ICU and hospital lengths of stay. These conditions are probably underrecognized at present. ICUs have the potential to serve as environments for neurologic teaching and research. PMID- 8420740 TI - Improvement of fecal fat excretion after addition of omeprazole to pancrease in cystic fibrosis is related to residual exocrine function of the pancreas. AB - Pancreatic function tests were performed in 11 adult cystic fibrosis (CF) patients with a fecal fat excretion of more than 10% during treatment with pancrease 2 capsules three times a day. These tests included urinary p aminobenzoic acid (PABA) excretion, fasting serum trypsin and pancreatic polypeptide (PP), and glucose and insulin in fasting and postprandial serum. Subsequently, the patients entered a double-blind placebo-controlled crossover study to assess the effect of gastric acid inhibition by 20 mg omeprazole on fecal fat excretion. Adjunct therapy with omeprazole resulted in a reduction of fecal fat excretion in patients with residual pancreatic function. This improvement showed significant positive correlations with urinary PABA excretion and the increase in serum PP after the meal (P < 0.02 and P < 0.05), but not with the other parameters studied. Therefore, the addition of omeprazole to pancrease is most successful in CF patients with residual pancreatic function, determined by urinary PABA excretion or incremental PP. PMID- 8420741 TI - Esophageal eosinophilia with dysphagia. A distinct clinicopathologic syndrome. AB - Small numbers of intraepithelial esophageal eosinophils (IEE) may be seen in 50% of patients with gastroesophageal reflux disease and occasionally in normal volunteers. High concentrations of IEE are rarely seen in either setting. During a two-year period we identified 12 adult patients with very dense eosinophil infiltrates in esophageal biopsies (defined as > 20 IEE/high-power field). Dysphagia was the presenting complaint in each, but no evidence of anatomical obstruction could be found. Endoscopic esophagitis was absent, but biopsy showed marked squamous hyperplasia and many IEE. Eleven patients had normal esophageal acid exposure on 24-hr pH monitoring. Esophageal manometry showed a nonspecific motility disturbance in 10 patients. For comparison, 90 patients with excess esophageal acid exposure on 24-hr pH monitoring were studied. Thirteen (14%) had motility disturbance, and 21 (23%) had dysphagia. Esophageal biopsies were devoid of IEE in 47 patients; none of the 43 with IEE had infiltrates as dense as those seen in the 12 study patients. The presence of high concentrations of IEE in esophageal biopsies from patients with dysphagia, normal endoscopy, and normal 24 hr esophageal pH monitoring represents a distinctive clinicopathologic syndrome not previously described. PMID- 8420742 TI - Link between Helicobacter pylori-associated gastritis and duodenal ulcer. AB - We examined the interrelationships among the degree of fundic mucosal atrophy, the prevalence of Helicobacter pylori in the gastric antrum, the gastric juice, and the duodenum with and without gastric metaplasia, in 20 duodenal ulcer patients and 20 non-duodenal ulcer patients. The detection rates of H. pylori in the antrum, the gastric juice, and the duodenum were significantly higher in duodenal ulcer patients (80%, 65%, and 60%) than in non-duodenal ulcer subjects (50%, 20%, and 5%). The frequency of H. pylori was significantly lower in the gastric juice (30%) and the duodenum (10%) in non-duodenal ulcer patients with antral H. pylori, compared with those in duodenal ulcer patients with antral H. pylori. All of seven patients with both gastric metaplasia and H. pylori infection in the duodenum had duodenal ulcer, whereas only 1 of 14 patients without either gastric metaplasia or H. pylori infection in the duodenum had duodenal ulcer. There was normal or mild atrophic mucosa in the fundus of duodenal ulcer patients with H. pylori in the antrum, whereas moderate or severe atrophic mucosa in non-duodenal ulcer patients with H. pylori gastritis. These results suggest that the preserved fundic mucosa, gastric metaplasia in the duodenum, and a greater load of H. pylori to the duodenum through the gastric juice may be prerequisites for the formation of duodenal ulcers. PMID- 8420743 TI - Pancreatic secretion in chronic alcoholics. Effects of acute alcohol or wine on response to a meal. AB - To gain further insight on the effects of alcohol on human pancreatic enzyme secretion, we tested the effects of a 12% (v/v) alcohol solution, wine, and a glucose solution added to a meal on trypsin output in duodenal aspirate of nonalcoholic volunteers and compared the results to those of chronic alcoholics. Plasma concentrations of gastrin, cholecystokinin, and pancreatic polypeptide were monitored pre- and postprandially. Similar blood alcohol concentrations were determined in nonalcoholics and alcoholics following wine and the alcohol solution. Nonstimulated trypsin output (basal) was higher in alcoholics but not significantly so when compared to nonalcoholics. However postprandial trypsin output, 2014 +/- 301 mg/5 hr was significantly greater in alcoholics (P < 0.05) compared to nonalcoholics 1271 +/- 118 mg/5 hr. Alcohol and wine when added to the meal significantly (P < 0.05) inhibited trypsin output in both groups. Basal and postprandial levels of gastrin and cholecystokinin were similar in nonalcoholics and alcoholics. Basal plasma pancreatic polypeptide levels were similar in both groups, but the postprandial increments in pancreatic polypeptide levels observed in nonalcoholics were not observed in alcoholics. We conclude that chronic alcoholics have increased postprandial pancreatic enzyme secretion, and that this secretion, as that of nonalcoholics, can be affected by alcohol or wine. The postprandial hypersecretion of enzymes in alcoholics is not related to increased plasma levels of cholecystokinin or gastrin. It is possible that the impaired release of pancreatic polypeptide may participate in the mechanism for increased pancreatic enzyme secretion in chronic alcoholics. PMID- 8420744 TI - Helicobacter pylori increases gastric antral juxtamucosal pH. AB - In order to examine the effect of Helicobacter pylori colonization on the gastric mucus microclimate, antral juxtamucosal pH was measured in 47 patients attending as out patients for upper gastrointestinal endoscopy. The mean pH in 28 patients negative for H. pylori was 6.40 +/- 0.24 compared to 6.88 +/- 0.16 in 19 patients who were positive (P < 0.0001). In six of seven patients who agreed to a second study, H. pylori was eradicated and the mean pH fell from 6.81 +/- 0.17 to 6.08 +/- 0.16 (P < 0.001). The pH remained high in the one patient who remained positive (6.8 and 7.0). This study provides the first in vivo evidence that H. pylori can increase the antral juxtamucosal pH and suggests that ammonia production by the organism is capable of altering gastric mucus microclimate to impair the normal negative feedback controlling gastrin release. This observation may explain the coexistence of relative hypergastrinemia and H. pylori colonization in duodenal ulcer patients. PMID- 8420745 TI - Helicobacter pylori in duodenal ulcer patients with idiopathic gastric acid hypersecretion. AB - Thirty-three consecutive patients with idiopathic gastric acid hypersecretion (defined as a basal acid output > 10.0 meq/hr with a normal fasting serum gastrin level and negative secretin stimulation test) who were being treated for duodenal ulcer disease and other acid-peptic disorders were evaluated for the presence of Helicobacter pylori by means of a rapid urease test. Fourteen patients had duodenal ulcer and 19 had other acid-peptic disorders (gastroesophageal reflux in 14, including six with Barrett's esophagus; four with nonulcer dyspepsia; and one with erosive gastritis). Helicobacter pylori was present in 12 of the 14 ulcer patients (86%) compared to only two of the 19 nonulcer patients (11%) (P < 0.0001). The distribution of basal acid output for patients with duodenal ulcer was similar to that for nonulcer patients, and no significant difference in the mean basal acid output was found among Helicobacter pylori-positive compared to Helicobacter pylori-negative patients. Seven of the duodenal ulcer patients with a basal acid output greater than 15.0 meq/hr were Helicobacter pylori-positive, suggesting that the organism can withstand even extreme levels of gastric acidity. In conclusion, this study demonstrates that the prevalence of Helicobacter pylori infection in patients with duodenal ulcer disease associated with idiopathic gastric acid hypersecretion is not different from a majority of ulcer patients with normal acid secretory profiles and offers additional evidence that extreme levels of gastric acid are not bactericidal for the organism. PMID- 8420746 TI - A neutrophil chemotactic factor present in H. pylori but absent in H. mustelae. AB - This study was designed to compare the capabilities of Helicobacter pylori and Helicobacter mustelae to generate neutrophil chemotactic activity (NCA) in vitro. H. pylori and H. mustelae were grown in parallel cultures under identical conditions. The cultures were washed and transferred to saline solution for 3 hr to avoid detecting nonspecific chemotactic activity from culture media. Supernatants were subjected to size-exclusion HPLC. All peaks from HPLC were collected and assayed for NCA. Peaks having significant NCA were subjected to gel electrophoresis. H. pylori generated 85.9 +/- 1.7% NCA compared to only 41.6 +/- 2.5% for H. mustelae (P < 0.001). The HPLC peak containing the highest NCA from H. pylori revealed a band on gel electrophoresis at approximately 10.5 kDa. This band was not present on gels from H. mustelae. We conclude that H. pylori produces a neutrophil chemotactic factor lacking from H. mustelae. This offers an explanation for the histologic difference between gastritides caused by these organisms. PMID- 8420747 TI - Study of Helicobacter pylori colonization of patches of heterotopic gastric mucosa (HGM) at the upper esophagus. AB - Helicobacter pylori (HP), known to cause active chronic gastritis, has primarily been found in gastric-type mucosa. Even in the duodenum, the organism was detected in islands of metaplastic gastric mucosa. HP has also been found in gastric metaplasia of Barrett's esophagus in 15-50%. The aim of our study was to determine: (1) the frequency with which HP is found on histopathological sections of heterotopic gastric mucosa (HGM) patch(es) at the upper esophagus, as compared to that of the stomach proper, and (2) the histopathological significance of infection in the HGM patches. From 63 patients with HGM patches at the upper esophagus, 48 patients were found to have concurrent adequate specimen from the stomach for modified Steiner's stain. In 22 patients (45.8%), pair sections from HGM and stomach were negative for HP. Of 26 patients (54.1%) HP-positive on sections from the antrum and/or body (both in 21 cases) nine patients (18.7%) demonstrated HP in the HGM patches. Whereas focal acute inflammatory changes on the H&E section of HGM was present in six patients, HP was detected in HGM only in one. Chronic inflammatory cell infiltration was detected in all nine HP positive HGM patches and in 37 of 39 HP-negative patches. A mixed acute and chronic inflammatory cell infiltration was found in five of these 37 patients. Our data demonstrate that HP infection of HGM patches at the upper esophagus is part of the HP gastritis and an independent colonization of HGM patches without gastric infection does not occur.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420748 TI - What is behind dyspepsia? AB - The first aim of the present study was to determine the cause of dyspepsia after negative conventional diagnostic work-up. In such patients, an extended diagnostic work-up was performed including esophageal pH monitoring and manometry, gastric and hepatobiliary scintigraphy, and lactose tolerance test. In 88 of 220 dyspeptic patients (mean age 49 years, range 17-87; 114 women) presenting to our gastroenterological outpatient department, a cause for dyspepsia was found by conventional work-up. Thirty-one of the remaining patients did not enter extended work-up because of minor symptoms. In 47 of 101 patients entering extended work-up, a diagnosis was established (21 endoscopy-negative gastroesophageal reflux disease, 11 gastric stasis, 6 biliary dyskinesia, and 5 lactase deficiency among them). A second aim of the study was to determine whether clusters of symptoms such as "gastroesophageal reflux-like," "dysmotility like," and "dyspepsia of unknown origin" reliably predict the groups of diseases suggested by these terms. This was not the case. In conclusion, in 40% of dyspeptic patients, a conventional diagnostic work-up led to a diagnosis that explained a patient's symptoms. After a negative conventional diagnostic work-up, an extended diagnostic work-up with functional tests yielded a possible explanation for their symptoms in 47% of patients. In such patients symptomatology was of little help for predicting the diagnosis. PMID- 8420749 TI - Differences in antigen expression between neoplastic and nonneoplastic gallbladder epithelium. An immunohistochemical study. AB - Immunoreactivity for a panel of 15 monoclonal antibodies (MAbs), which are known to react with different gastrointestinal tumor antigens, was assessed in formalin fixed paraffin-embedded sections that were prepared from cholecystectomy specimens obtained from Mexican patients. Each case was classified histologically into one of the following groups: (1) invasive adenocarcinoma (N = 21), (2) high grade dysplasia (carcinoma in situ) (N = 2), (3) low-grade dysplasia (N = 4), hyperplasia (4) (N = 15), and (5) chronic cholecystitis (N = 10). Significant differences (P < 0.05) were identified among the five histopathologic groups in the proportion of epithelial cells demonstrating immunoreactivity with MAbs to Lewisb; Lewis(a); sialylated Lewis(a); sialylated Lewis(a) and Lewis(a); Y antigen; H antigen; X antigen; X-like antigen; 200-kDa protein of CEA; 180-, 160 , 50-, 40-kDa proteins of CEA; 30- to 37-kDa protein; and an undefined antigen identified by MAb 99-57, with invasive carcinoma more frequently being positive as compared to nonneoplastic (hyperplasia, chronic cholecystitis) epithelium. Significant differences were also observed among the five histopathologic groups (P < or = 0.0005) in the proportion of epithelial cells demonstrating immunoreactivity with MAbs to Y antigen and the 20- to 50-kDa glycoprotein. However, with these two antibodies immunoreactivity was more frequently found in nonneoplastic epithelium rather than in invasive carcinomas. No significant differences in immunoreactivity were detected among the different histologic groups with MAb to blood group B antigen, types 1 and 2. This study demonstrates that cellular antigens are both developed and lost during the process of neoplastic transformation in the gallbladder.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420750 TI - High vesicular cholesterol and protein in bile are associated with formation of cholesterol but not pigment gallstones. AB - To examine the differentiating parameters between cholesterol and pigment gallstones, we compared the nucleation times, concentrations of biliary lipid and protein, and the distribution of vesicular cholesterol in gallbladder bile of 16 patients with cholesterol, eight patients with black pigment gallstones, and nine gallstone-free control patients. Cholesterol monohydrate crystals were present in the fresh bile of only the cholesterol gallstone group. The nucleation time was significantly faster in the cholesterol stone group (3.3 +/- 3.2 days) than in the other two groups (pigment stone: 15.8 +/- 6.6, control: 16.9 +/- 5.7). The cholesterol saturation indices and the distribution of vesicular cholesterol were significantly higher in the cholesterol gallstone group than those in the other two groups. The total biliary protein concentration was significantly (P < 0.01) higher in the cholesterol gallstone group [2.57 +/- 1.91 (SD) mg/ml] than that in the black pigment stone group (1.09 +/- 0.59). All parameters in patients with black pigment gallstone were essentially similar to the controls. We conclude that the presence of cholesterol crystals, rapid nucleation time, high vesicular cholesterol distribution, elevated cholesterol saturation index, and high protein concentration are associated with cholesterol gallstones but not with black pigment gallstones. PMID- 8420751 TI - Prostaglandin E2 stimulates ion transport in prairie dog gallbladder. AB - The effects of prostaglandins, and specifically prostaglandin E2, on gallbladder ion transport were examined in the prairie dog. Gallbladders were mounted in an Ussing chamber and baseline short-circuit current, potential difference, and tissue resistance were measured. Addition of arachidonic acid (10(-4) M, mucosal surface) produced sustained elevations in short-circuit current and potential difference (P < 0.05), with mild reductions in resistance. In a second set of tissues, indomethacin exposure (10(-6) M) resulted in a significant (P < 0.02) decrease in short-circuit current and potential difference, with an increase in resistance. Subsequent addition of prostaglandin E2 (10(-7) M, serosal surface) fully reversed these changes and led to a significant increase in short-circuit current and potential difference (P < 0.001) with a return of resistance to baseline values. These findings suggest that endogenous prostaglandins mediate gallbladder ion transport. PMID- 8420752 TI - Gangliocytic paraganglioma, a rare cause of upper gastrointestinal bleeding. Endoscopic ultrasound findings presented. PMID- 8420753 TI - Lusorian artery lesion as rare cause of severe upper gastrointestinal tract bleeding. AB - A patient in an intensive care unit experienced severe esophageal bleeding caused by erosion of a lusorian artery. The lusorian artery is a rare variant of the right subclavian artery. It originates in the descending aortic arch and crosses behind the esophagus to the right, sometimes generating esophageal compression. The patient's condition required respirator therapy and placement of a duodenal tube. At the point of crossing over of the lusorian artery and the esophagus, the duodenal tube caused esophageal necrosis, leading to erosion of the lusorian artery. This resulted in extensive esophageal bleeding, which at last required surgical intervention. To attain proper treatment and to avoid unnecessary diagnostic and therapeutic approaches, a lusorian artery lesion has to be included in the differential diagnosis of upper gastrointestinal bleeding. PMID- 8420755 TI - Gastrointestinal involvement in a woman with dyskeratosis congenita. AB - Dyskeratosis congenita (DC) is a rare hereditary condition with characteristic dermatological findings and is frequently associated with the development of pancytopenia. Involvement of the gastrointestinal tract and liver occurs in a significant proportion of patients. Although the disorder is transmitted in an X linked fashion, several affected females have been described. We report a case of DC involving a woman in whom gastrointestinal symptoms were prominent, primarily diarrhea, gastrointestinal hemorrhage, and ascites. Portal hypertension was evident by the presence of splenorenal varices. Extensive evaluation failed to disclose an etiology for either her colitis or portal hypertension. The literature concerning gastrointestinal involvement in DC is reviewed and a possible association with noncirrhotic portal hypertension is suggested. PMID- 8420754 TI - Impaired mononuclear phagocyte function in patients with severe acute pancreatitis: evidence from studies of plasma clearance of trypsin and monocyte phagocytosis. AB - Activated proteases in plasma are complexed by alpha 2-macroglobulin. Although the complexes retain peptidase activity, they are usually eliminated promptly by mononuclear phagocytes. In severe acute pancreatitis, almost 30% of plasma alpha 2-macroglobulin becomes complexed, suggesting impaired clearance. In the present study, plasma [methyl-14C]trypsin clearance and monocyte phagocytosis were investigated. Attacks complicated by major organ-system failure, pancreatic pseudocyst, abscess, or necrosis were graded severe (median Ranson score 5.5). Plasma [methyl-14C]trypsin half-life was significantly increased in severe attacks (N = 7, median 21.1 min), compared to mild attacks (N = 14, median 15.4 min, P < 0.05) and healthy controls (N = 4, median 10.8 min, P < 0.02). Monocyte phagocytosis was significantly lower in severe attacks (N = 9, median 3.6%) compared to mild attacks (N = 20, median 20.8%, P < 0.01) and healthy controls (N = 8, median 26.9%, P < 0.01). Plasma [methyl-14C]trypsin half-life and monocyte phagocytosis were significantly inversely correlated (r = -0.51, P < 0.01). Impaired clearance of circulating trypsin in acute pancreatitis is potentially deleterious but may be reversed by stimulating mononuclear phagocytes. PMID- 8420756 TI - Familial adenomatous polyposis (Gardner's syndrome) and thyroid carcinoma. A case report and review of the literature. AB - The case history of a 24-year-old woman with Gardner's syndrome [familial adenomatous polyposis (FAP)] and papillary thyroid carcinoma is presented, representing the 37th report of this association. Although FAP is transmitted as an autosomal dominant trait with similar penetrance in both sexes, thyroid carcinoma has been found almost exclusively in women (94.3%). The majority have been papillary carcinomas (88.5%), which have become apparent during the third decade (average 23.6, range 16-40 years). Most (55.5%) thyroid carcinomas have been discovered 1-17 years after FAP was identified, although some have been found before (29.6%), or at the same time (14.8%) FAP was diagnosed. Multicentric papillary carcinomas have been reported in 64% (14 of 22) of FAP patients, a frequency at least twofold greater than usual. Although papillary carcinoma found before age 30 (as it was in most patients with FAP) typically has an excellent prognosis, one patient with FAP developed distant metastases from thyroid carcinoma and a 28-year-old woman's death was attributed to papillary carcinoma. The high frequency of multicentric papillary thyroid carcinoma in young patients with FAP and the potential for metastases and death due to thyroid carcinoma warrant aggressive diagnostic screening at regular intervals with neck palpation, ultrasonography, and if necessary, fine-needle aspiration biopsy. When thyroid carcinoma is found, total or near-total thyroidectomy should be considered because of the tumor's high likelihood of being multifocal.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420757 TI - Effect of cysteamine on insulin release and exocrine pancreatic secretion in vitro. AB - Cysteamine is known to deplete somatostatin from pancreatic D cells. In the isolated perfused rat pancreas we investigated its effects on somatostatin and insulin release as well as exocrine pancreatic secretion in the presence of 16.7 mM glucose and 180 pM CCK-8. At a concentration of 0.1 mM, cysteamine had no significant effect on pancreatic endocrine and exocrine functions. At 10 mM, however, cysteamine released somatostatin (380 +/- 70 vs 100 +/- 20 fmol/20 min), inhibited insulin output (890 +/- 120 vs 13210 +/- 3260 mu units/20 min) and reduced exocrine pancreatic secretion (volume: 12 +/- 2 vs 20 +/- 2 microliters/20 min; lipase: 31 +/- 3 vs 60 +/- 7 units/20 min). We conclude that the complex changes induced by cysteamine are consistent with a physiological role of endogenous somatostatin in the regulation of insulin release. The reduction of exocrine pancreatic secretion, however, was at least in part, if not completely, mediated via the insuloacinar axis rather than a direct effect of cysteamine-released somatostatin on pancreatic acinar cells. PMID- 8420758 TI - Bile-induced acute pancreatitis in cats. Roles of bile, bacteria, and pancreatic duct pressure. AB - The relationship between pancreatic duct pressure, bile, bacterial infection of bile, and the development of acute pancreatitis was studied in a feline model. The cat main pancreatic duct was perfused from the head to the tail of the gland with bile and/or Escherichia coli bacteria at "basal" pancreatic duct pressure (< 10 cm H2O) and at pancreatic duct pressure in the upper physiologic range (45 cm H2O). Sterile bile and sterile control solution produced no inflammatory alterations at either pressure. Infected control solution caused a mild acute edematous pancreatitis under low and high pressure. Infected bile caused a severe acute edematous pancreatitis at basal duct pressure; at high pressure, additional focal acinar necrosis was observed in the majority of animals. Infected bile was found to raise basal pancreatic duct pressure by 30%. The other test solutions, which caused only mild inflammatory alterations of the pancreas, did not alter duct pressure. We conclude: (1) Bacterial infection is important for the initiation of acute pancreatitis in this model. (2) High physiologic duct pressure may result in the conversion of nondestructive forms of the inflammation to acinar necrosis. (3) Infected bile-induced increase in duct pressure is likely to result from compression of the duct lumen by the inflammatory edema of the gland. PMID- 8420759 TI - Relation of symptoms to impaired stomach, small bowel, and colon motility in long standing diabetes. AB - Stomach, intestinal, and colonic transit were measured in males with insulin requiring diabetes of greater than 10 years' duration to compare with symptoms and to estimate the medical significance. For all diabetics only the symptom constipation correlated with the appropriate regional delayed transit. Diabetics with delayed transit in any region, however, had more overall gastrointestinal symptoms. Diabetics with delayed transit had disease of significantly longer duration than those without delay. Delayed transit was common in the diabetics selected for study with 21 of 54 stomachs, 10 of 20 small intestines, and 14 of 20 colons showing impairment. Of 35 diabetics with impaired transit at one or more locations, only seven were judged of medical importance and five of these responded to treatment. In this study, delayed transit was frequent; in the one fifth requiring management, the symptoms related closely to the region impaired. PMID- 8420760 TI - Reduced postprandial blood glucose levels in recently diagnosed non-insulin dependent diabetics secondary to pharmacologically induced delayed gastric emptying. AB - In a previous study we demonstrated that patients with recently diagnosed non insulin-dependent diabetes mellitus (NIDDM) had significantly increased gastric emptying rates of glucose solutions compared with those of nondiabetic sex- and age-matched controls. This finding of rapid gastric emptying contrasts with the delayed gastric emptying often exhibited as a late manifestation of diabetes mellitus that is attributed to autonomic neuropathy. The purpose of this study was to determine, in seven of the patients previously studied, whether (1) an intravenous infusion of cholecystokinin-8 (CCK-8) would delay the gastric emptying of a liquid glucose meal and, if so, (2) whether the delay in gastric emptying would result in reduced postprandial blood glucose concentrations due to prolongation of the absorption of the glucose in the liquid meal. Each patient underwent two separate gastric emptying studies, one during a saline infusion and one during a CCK-8 infusion. Blood samples were obtained at 15-min intervals for measurement of glucose, insulin, CCK-8, and gastric inhibitory polypeptide (GIP) concentrations. The average gastric half-emptying time was 41 min with the saline infusion and 94 min with the CCK-8 infusion (P = 0.0042). The average glucose concentration over the 2-hr period following glucose ingestion was 17.1 mmol/liter with the saline infusion and 14.0 mmol/liter with the CCK-8 infusion (P = 0.0073). The average glucose excursion value over the 2-hr period was reduced from 5.6 mmol/liter to 3.7 mmol/liter with the CCK-8 infusion (P = 0.0550).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420761 TI - Endosonography and cytology in diagnosing and staging pancreatic body and tail carcinoma. Preliminary results of endosonographic guided puncture. AB - Endosonography was performed in diagnosing and staging pancreatic body and tail carcinoma in two patients. In the first case endoscopy, abdominal ultrasound, and computed tomography were nondiagnostic in diagnosing the origin of submucosal gastric abnormalities. Endosonography diagnosed a pancreatic tail carcinoma with submucosal gastric involvement, and this was confirmed by endosonographic-guided cytology. Fundus varices due to segmented splenic vein involvement were found. Surgery was not recommended due to the advanced disease. In the second case pancreatic body carcinoma was diagnosed by ERCP and computed tomography. Transcutaneous ultrasonographic-guided cytological puncture confirmed the diagnosis. Endosonography revealed additional information of segmental portal hypertension with fundic varices due to splenic vein involvement. Autopsy confirmed the endosonographic diagnosis. PMID- 8420762 TI - Infectious complications of endoscopic procedures in bone marrow transplant recipients. AB - The prevalence of clinically relevant bacteremia after endoscopic procedures in bone marrow transplant recipients was assessed retrospectively. Bacteremia, within 24 hr of procedure, was defined as positive blood cultures, while hypotension and temperature greater than 38 degrees C were taken as possible indicators of bacteremia. Sixty-seven procedures were performed in 53 endoscopic sessions (upper endoscopy 37, flexible sigmoidoscopy 7, upper endoscopy + flexible sigmoidoscopy 8, colonoscopy 1). Twenty-five endoscopic sessions were performed in patients receiving broad-spectrum antibiotics and 28 sessions in patients not receiving antibiotics. Both groups were comparable with respect to patient characteristics, procedures performed, and immune status. No patient in either group developed hypotension. One patient developed fever after flexible sigmoidoscopy; no source of fever was identified. We conclude that: (1) there were no episodes of clinically relevant bacteremia attributable to endoscopic procedures, and (2) not all bone marrow transplant recipients require routine antibiotic prophylaxis prior to endoscopic procedures. PMID- 8420763 TI - Pneumatic dilatation or esophagomyotomy treatment for idiopathic achalasia: clinical outcomes and cost analysis. AB - The choice between pneumatic dilatation and surgical esophagomyotomy as the initial treatment for achalasia is controversial. The aims of this study were to determine the long term clinical outcome and costs of treating achalasia initially with pneumatic dilatation as compared to esophagomyotomy. Of 123 patients undergoing an initial pneumatic dilatation for achalasia at our institution from 1976 to 1986, 71 (58%) received no further treatment for achalasia during a mean follow up of 4.7 +/- 2.8 years. Only 15 of these 123 patients (12%) eventually underwent surgical esophagomyotomy (two for perforation during pneumatic dilatation, 13 for persistent or recurrent symptoms). The degree of dysphagia at follow up was improved to a similar degree in patients treated with an initial pneumatic dilatation as compared to patients treated with an initial esophagomyotomy. Patients with age > or = 45 years at time of initial pneumatic dilatation had fewer subsequent treatments for persistent or recurrent symptoms and had less dysphagia on follow up as compared to patients < 45 years. Subsequent pneumatic dilatations to treat persistent or recurrent symptoms were less beneficial than an initial pneumatic dilatation. The cost of esophagomyotomy was 5 times greater than the cost of pneumatic dilatation. When costs were analyzed to include subsequent treatments of symptomatic patients, the total expectant costs of treating with an initial esophagomyotomy remained 2.4 times greater than treating with an initial pneumatic dilatation. This study suggests that an initial pneumatic dilatation will be the only treatment needed for the majority of patients with achalasia. A treatment regimen starting with pneumatic dilatation has less overall costs than starting with esophagomyotomy. For each subsequent pneumatic dilatation, however, the clinical benefit leans toward surgery. PMID- 8420764 TI - Fasting and postprandial mechanisms of gastroesophageal reflux in children with gastroesophageal reflux disease. AB - In order to define the mechanisms of gastroesophageal reflux (GER) in children, we performed simultaneous intraluminal esophageal motility and pH studies in 24 children with symptomatic reflux and abnormal prolonged pH probe study, ten (group A) without endoscopic and histologic esophagitis, 14 (group B) with endoscopic and histologic esophagitis. Median (ranges) age (years) was 5.0 (6 months-10 years) and 3.0 (6 months-12 years), respectively. Recordings were done for 1 hr before and 1 hr after feeding apple juice (15 ml/kg; pH 4.0). All episodes of GER in group A patients and 77.1% in group B patients were accounted for by abrupt transient lower esophageal sphincter (LES) relaxation (TLESR); 22.9% of reflux events in group B patients occurred during gradual drifts of LES pressure (LESP) to undetectable levels. Esophageal refluxate exposure (mean percentage time with esophageal pH < 4.0), the rate of TLESR (number of episodes/hr), and the percentage of TLESRs associated with reflux significantly increased in the fed period both in group A (18.5 +/- 5.4%, 6.2 +/- 2.65, 87.1%) and in group B (29.7 +/- 6.5, 7.8 +/- 3.05, 84.9%) as compared to the fasting state (group A: 10.8 +/- 3.9, 3.9 +/- 3.17, 46.1%; group B: 16.1 +/- 2.6, 4.14 +/ 3.06, 55.17%) (p < 0.01). The rate of LESP drifts (number of episodes/hr) was also significantly higher postprandially (4.85 +/- 1.24 vs 1.8 +/- 0.9, p < 0.01); furthermore there was a postfeeding increase of the LESP drift percentage associated with reflux (79.41% vs 46.15%, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420765 TI - Low-proof alcoholic beverages and gastroesophageal reflux. AB - Previous studies have demonstrated an increased gastroesophageal reflux after the ingestion of high-proof alcoholic beverages in normal subjects. Data on gastroesophageal reflux with usual amounts of low-proof alcoholic beverages are not available. The effect of white wine (7.5% v/v, pH 3.2) and beer (7.0% v/v, pH 4.5) was compared with water, a nonalcoholic beverage of pH 3.2, and an ethanol solution (7.5% v/v, pH 7.6) using ambulatory pH measurement in healthy volunteers. The fraction of time at pH < 4 in the first hour after ingestion of 300 ml white wine (median 13.2%) was significantly increased compared with beer (3.6%; P < 0.01), water (0.9%; P < 0.001), ethanol (1.3%; P < 0.001), and the nonalcoholic beverage (0.9%; P < 0.05). Beer provoked significantly more gastroesophageal reflux than water (P < 0.01). It is concluded that white wine and beer induce gastroesophageal reflux, which is neither related to their ethanol content nor to their pH. The mechanism for this effect remains to be identified. PMID- 8420766 TI - Detection by scanning electron microscopy of a distinctive esophageal surface cell at the junction of squamous and Barrett's epithelium. AB - Metaplastic columnar epithelium replaces the normal squamous epithelium in Barrett's esophagus. We characterized the surface epithelial cells of the junction between squamous and Barrett's epithelium using scanning electron microscopy and light microscopy. In four biopsy specimens from the squamous Barrett's junction in three patients, we found a distinctive cell type having features intermediate between those of squamous and columnar epithelium. Its distinguishing characteristic is the presence on its surface of two disparate structures not normally present on the same cell in the gastrointestinal tract: microvilli (a scanning electron microscopy feature of glandular epithelium) and intercellular ridges (a scanning electron microscopy feature of squamous mucosa). The surface characteristics of this newly recognized cell were strikingly similar to those of cells found in the transformation zone of the uterine cervix, an area in which squamous epithelium physiologically replaces columnar epithelium. We also examined 28 biopsies of the gastroesophageal junction area from 14 patients with and without a history of heartburn but with no evidence of Barrett's esophagus. None of these biopsies showed the distinctive cell. We hypothesize that this distinctive cell represents an intermediate step in either the development or the healing of Barrett's epithelium, during which surface characteristics of two different cell types, columnar and squamous, coexist on the same cell. PMID- 8420767 TI - [Fibrin agglutination of thyroid gland cysts after fine needle puncture]. AB - Tolerance to and efficacy of a fibrin adhesive injected into thyroid cysts after their evacuation by fine needle puncture was investigated in 60 consecutive patients (52 women, 8 men; mean age 50 [18-82] years) with solitary thyroid cysts. The patients were randomly separated into two groups of 30 each. In group A patients the cysts were completely emptied by fine needle puncture under ultrasound control. In group B patients a fibrin adhesive (0.8-1.4 ml) was subsequently injected. 25 of 30 patients in group A had recurring cysts, twelve of which were larger than the original one. There were 13 recurrences in group B (P < 0.005), but only one was of a larger volume. In each group three patients reported a brief period of pain after the evacuation procedure. The described technique has proven to be reliable and well tolerated. Cysts obliterated by fibrin adhesive recurred less frequently and had a smaller volume than those only evacuated by needle puncture. PMID- 8420768 TI - [Acute pancreatitis and stomach wall necrosis caused by cholesterol embolisms]. AB - A 60-year-old man was hospitalized because of a sudden onset of severe pain in the epigastrium and haematemesis. Acute pancreatitis was diagnosed on the basis of an increased serum amylase concentration (642 U/l). Abdominal ultrasound and computed tomography demonstrated a necrotic zone with central liquefaction in the tail of the pancreas adjoining the stomach wall. Gastroscopy revealed as source of the bleeding an extensive mucosal necrosis at the greater curvature of the stomach. At laparotomy, partial resection of the pancreas, gastrectomy and splenectomy were performed. Histological examination of the resected specimens showed multiple cholesterol emboli in the small arteries of the pancreas and the gastric submucosa. PMID- 8420769 TI - [Hypersalivation as a leading symptom of neoplastic meningiosis in highly malignant non-Hodgkin's lymphoma]. AB - Partial remission of a centroblastic non-Hodgkin's lymphoma, clinical stage IV A, in a 79-year-old man was achieved by six courses of chemotherapy with epirubicin, cyclophosphamide and vincristine. The only residual finding was a palpable small cervical lymphoma. After a treatment pause of about 6 weeks increasing hypersalivation set in which ultimately made food intake impossible and led to a breakdown in the patient's general state. Findings in the region of the head, neck, throat and the base of the skull were unremarkable, but cerebrospinal fluid contained 1300/3 cells, almost all of them lymphoblasts. After five intrathecal injections of at first 15 mg methotrexate and 4 mg dexamethasone each, followed by five more with 40 mg cytarabine added to them, the CSF cell count became normal. At the same time salivation clearly decreased and food intake became once again possible. The patient died 5 months later from hypercalcaemia due to osseous infiltrations. Until his death there was no recurrence of the hypersalivation as the cardinal sign of meningeal carcinomatosis. PMID- 8420770 TI - [Borderline between ambulatory and stationary medicine in the example of diabetes]. PMID- 8420771 TI - [Nevus cell nevi, solar exposure and risk of melanoma]. PMID- 8420772 TI - [Cholesterol in food and serum]. PMID- 8420773 TI - [Nitinol stent in esophageal carcinoma]. PMID- 8420774 TI - [Healing of chronic psoriasis vulgaris while using omeprazole]. PMID- 8420775 TI - [Illusion, worm or artefact?]. PMID- 8420776 TI - [Clinical experiences with nocturnal intermittent peritoneal dialysis]. AB - Nocturnal intermittent peritoneal dialysis (NIPD) was performed in 56 patients (37 men, 19 women; mean age 44 [18-77] years, 18 of them diabetics). The NIPD protocol laid down 6-10 automatic changes of 1.5-2.3 1 dialysate (mean glucose concentration: 1.9%) each night. Clinical and biochemical tests were performed three times, at least 4 weeks apart (the first one after 2 months of stable NIPD). 36 of the 52 patients under the age of 60 were working. Mean hospital stay (excluding catheter implantation and training) was 5 days yearly. Mean peritonitis rate was one episode every 54 months. Because of technical problems a change from peritoneal to haemodialysis became necessary in four patients. Clinical and biochemical findings were: creatinine 877 +/- 269 mumol/l, urea 20 +/- 5.2 mmol/l, potassium 4.5 +/- 0.6 mmol/l, calcium 2.3 +/- 0.2 mmol/l, phosphate 1.7 +/- 0.5 mmol/l, total protein 69 +/- 7 g/l, cholesterol 6.8 +/- 1.7 mmol/l, and triglycerides 2.6 +/- 1.4 mmol/l. These data show that NIPD is an effective method of dialysis which--in comparison with other techniques--has several advantages. Peritonitis and technical failure are rare. PMID- 8420777 TI - Quantification of technetium-99m lung radioactivity from planar images. AB - Six methods of quantifying technetium-99m lung uptake from planar gamma camera images were evaluated. Camera sensitivities, the broad beam attenuation coefficient and build-up factors were derived from suitable phantom measurements. The accuracy of the methods was evaluated by quantifying the lung uptake of 99mTc macroaggregated albumin (MAA) in ten patients and assuming complete trapping by the lung of the known activity of injected MAA particles. Three methods based on published techniques which related the count rate from an image of a lung to that from a lung phantom were the least accurate, producing lung activities which were typically about 70% of the injected activity. Of the other three techniques, the depth-dependent build-up factor method was slightly more accurate than the geometric mean and the depth-independent build-up factor methods, producing average values (+/- SD) of lung activity which were 100% +/- 3%, 106% +/- 3% and 101% +/- 5%, respectively, of the injected activity. To measure lung uptake, all of these latter three methods required an attenuation correction with a flood source transmission scan, and therefore their accuracy was affected by the variation in the activity distribution and attenuation across an image of the thorax. PMID- 8420778 TI - Thyroidal handling of radioiodine in sea level residents exposed to hypobaric hypoxia. AB - In the present investigation thyroidal accumulation of radioiodine and its release were assessed by direct testing of thyroid function using radioactive iodine, in vivo, in sea level residents intermittently exposed to hypobaric hypoxia. Thyroidal accumulation of radioiodine and its turnover were examined daily for 14 days. Twelve healthy human male volunteers were divided into three groups, with an equal number of individuals in each group. A decompression chamber was used to expose each group of subjects to hypoxic conditions at a simulated altitude of 3810 m for 8 h/day for 14 days. An oral dose of 25 mu Ci iodine-131 was administered to each individual of the first group immediately before the initiation of intermittent hypoxia. The second group of subjects received a tracer dose at the beginning of the 4th day of the 14 days, intermittent exposure to hypoxia, while the third group received the tracer dose 1 week after the completion of the exposure. Control studies were carried out on the subjects before they were subjected to the experimental conditions. Thyroidal accumulation of 131I in experimental subjects during the hypoxic state and in the post-hypoxic state was higher than in the control studies. The pattern of accumulation during exposure to hypoxia and in the post-hypoxic state showed multiple peaks of radioactive iodine uptake (PRAIU), a unique feature. The multiple PRAIU by the thyroid in experimental subjects were sharp and of short duration, reflecting an increased rate of 13I release from the thyroid. Control subjects had a single PRAIU by the thyroid 24 h after the administration of tracer. PMID- 8420779 TI - A comparison of rest sestamibi and rest-redistribution thallium single photon emission tomography: possible implications for myocardial viability detection in infarcted patients. AB - Thirty patients (26 men, 4 women, mean age 61 +/- 8 years) who had suffered myocardial infarction 15 +/- 6 months previously, were submitted to (1) standard stress-redistribution thallium-201 single photon emission tomography (SPET), (2) rest-redistribution 201T1 SPET and (3) stress-rest technetium-99m sestamibi SPET. Uptake modifications in relation to exercise-induced defects were evaluated in a total of 390 myocardial segments. Tracer uptake was scored as normal (=0), mildly reduced (=1), apparently reduced (=2), severely reduced (=3) or absent (=4). Comparison of stress studies failed to show any statistical difference (58% segmental abnormalities with sestamibi vs 61% with thallium). Uptake abnormalities (score 1-4) were detected in 55% of the segments with sestamibi, 55% with standard thallium redistribution, 55% with early imaging after thallium injection at rest and 54% with 3-h delayed rest imaging (P = NS). Absence of tracer uptake (score = 4) under resting conditions was recorded in 75 (19%) segments with standard 201T1 redistribution, 75 (19%) with rest sestamibi, 70 (18%) with rest 201T1 imaging and 62 (16%) with rst-redistribuion 201T1 (P < 0.05 vs other imaging modalities). Thus, 3-h delayed rest thallium imaging detected reversibility of uptake defects in a significantly higher number of myocardial segments. This finding might have important implications for both tracer and technique selection when myocardial viability is the main clinical issue. PMID- 8420780 TI - Improvement of brain single photon emission tomography (SPET) using transmission data acquisition in a four-head SPET scanner. AB - Attenuation coefficient maps (mu-maps) are a useful way to compensate for non uniform attenuation when performing single photon emission tomography (SPET). A new method was developed to record single photon transmission data and a mu-map for the brain was produced using a four-head SPET scanner. Transmission data were acquired by a gamma camera opposite to a flood radioactive source attached to one of four gamma cameras in the four-head SPET scanner. Attenuation correction was performed using the iterative expectation maximization algorithm and the mu-map. Phantom studies demonstrated that this method could reconstruct the distribution of radioactivity more accurately than conventional methods, even for a severely non-uniform mu-map, and could improve the quality of SPET images. Clinical application to technetium-99m hexamethyl-propylene amine oxime (HMPAO) brain SPET also demonstrated the usefulness of this method. Thus, this method appears to be promising for improvement in the image quality and quantitative accuracy of brain SPET. PMID- 8420781 TI - The role of thallium-201 single photon emission tomography in the investigation and characterisation of brain tumours in man and their response to treatment. AB - The aim of this study was to characterise brain tumour type and treatment response in relation to the uptake of thallium-201. 201T1 single photon emission tomography (SPET) was performed in 58 patients with brain tumours. Fifty-six patients were utilised for the statistical comparison of the early and delayed 201T1 indices expressed as the ratio of tumour to contralateral cerebral hemisphere uptake. The retention index of 201T1 in the tumour tissue calculated from the early and delayed scans was also analysed. Furthermore, in 56 patients with 58 brain tumours, a comparison was made of the diagnostic value of high 201T1 uptake and gadolinium diethylene triamine penta-acetic acid (Gd-DTPA) enhancement on MRI scans. Although high 201T1 uptake was observed in viable malignant gliomas, brain metastases, meningiomas and malignant teratoma, the viable malignant gliomas could not be differentiated from brain metastases and extracerebral tumours by means of 201T1 indices. 201T1 SPET failed to diagnose a viable ring-enhanced tumour with a thin rim and small tumours of less than 1.5 cm in diameter visualised by Gd-DTPA-enhanced MRI. In spite of this, 201T1 SPET appears to be effective for determination of the malignant viability of tumours. PMID- 8420782 TI - 99mTc-DTPA aerosol for same-day post-perfusion ventilation imaging: results of a multicentre study. AB - A multicentre study was performed in an attempt to evaluate a submicronic technetium-99m diethylene triamine penta-acetic acid aerosol generated by a newly developed delivery system, the aerosol production equipment (APE nebulizer), for same-day post-perfusion ventilation imaging in patients with clinically suspected pulmonary embolism. Quantitative comparison between the DTPA aerosol and krypton gas demonstrated a close correlation with respect to regional pulmonary distribution of activity and peripheral lung penetration (n = 14, r = 0.94, P < 0.001 and r = 0.75, P < 0.0025, respectively). In 169 consecutive patients, DTPA aerosol images performed immediately following perfusion (inhalation scan I) were compared to those carried out on the next day (inhalation scan II) with respect to image quality and assessment of perfusion-ventilation matches or mismatches. Agreement between inhalation scans I and II with respect to perfusion defects matched or mismatched to ventilation was found in 166/169 (98%) studies. The image quality of inhalation scan I was equal to that of scan II in 72%; inhalation scan I was superior in 11% of cases, while scan II was superior in 17%. This submicronic 99mTc-labelled DTPA aerosol is well suited for fast same day post-perfusion ventilation imaging in patients with clinical suspicion of pulmonary embolism. PMID- 8420783 TI - Application of carbon-11 labelled nicotine in the measurement of human cerebral blood flow and other physiological parameters. AB - Using positron emission tomography (PET), we measured the regional cerebral blood flow (rCBF) in five normal human subjects after intravenous injection of carbon 11 labeled (R)nicotine. The rCBF of the same subjects was measured by PET using the C15O2 inhalation steady-state method. The distribution of 11C activity in the brain after injection of 11C-(R)nicotine was almost equivalent to the CBF image obtained with the C15O2 inhalation stead-state method. The kinetics of 11C (R)nicotine in the brain was analysed using a two-compartment model consisting of vascular and brain tissue compartments. The rCBF values obtained with 11C (R)nicotine were higher than with C15O2 gas. The relatively long fixed distribution of 11C-(R)nicotine with a short uptake period allows stimulation studies by measurement of CBF to be performed with better photon flux and a longer imaging time than are possible with H215O. PMID- 8420784 TI - Postoperative assessment of cerebral blood flow in subarachnoid haemorrhage by means of 99mTc-HMPAO tomography. AB - Regional hypoperfusion is a very frequent complication of subarachnoid haemorrhage (SAH), being related to vasospasm in the majority of cases. Twenty six patients who were admitted for SAH underwent follow-up with technetium-99m hexamethylpropylene amine oxime single photon emission tomography (SPECT) 3 and 8 days after surgery. Fifteen patients of these had one more examination 15 days after surgery. The degree of hypoperfusion was quantified using an index of asymmetry which allow the comparison of two symmetrical regions of interest (ROIs) on the transaxial slice which presented the greatest perfusion defect. Comparison of CT data, transcranial Doppler data and clinical signs with the perfusion as quantified by 99mTc-HMPAO SPECT indicates that a difference in counts of less than 10% between the two symmetrical ROIs is of no diagnostic value. Follow-up of the brain perfusion clearly shows that the most pronounced hypoperfusion was observed just after surgery, with progressive normalization at 8 and 15 days after surgery. 99mTc-HMPAO SPECT performed 8 days after surgery allows prediction of the clinical outcome. For these reasons, 99mTc-HMPAO SPECT, which is the only method for follow-up of cerebral perfusion in routine clinical practice, should be the first examination to be performed after surgery in patients with SAH. PMID- 8420785 TI - Protocols for selection of cardiac radionuclide studies for use as a data base of normal studies and typical patterns of diseases. COST B2 Working Group II, in association with relevant working/task groups of the European Association of Nuclear Medicine and the European Society of Cardiology. AB - A data base of clinical studies is required for quality assurance of software used for analysis of radionuclide cardiac imaging procedures. Studies used must be rigorously validated in terms of both the clinical condition of the patient undergoing the procedure and the imaging protocol used. Selection protocols for the creation of a software phantom data base of normal studies and three typical patterns of cardiac disease--recent transmural myocardial infarction, isolated myocardial ischaemia and dilated cardiomyopathy--have been developed by the Cardiac Working Group of the European COST B2 project in association with the Cardiac Task Group of the European Association of Nuclear Medicine and the Working Group on Nuclear Cardiology of the European Society of Cardiology. These protocols include criteria for the interpretation of qualitative and quantitative non-radionuclide data. Compliance of the clinical data with the selection criteria will have to pass scrutiny by an international team for each study used as a software phantom. The radionuclide studies encompass stress and rest myocardial perfusion studies (planar and single photon emission tomography) using thallium-201 and technetium-99m methoxyisobutylisonitrile and rest gated blood pool studies. Methods for acquisition of data are defined for each type of study and for each individual study a portfolio of all clinical data is established. A pilot study is required to investigate the problems and logistics of distributing clinical radionuclide studies between a range of computers and institutes, and to ascertain the procedures necessary for analytical comparison of the results obtained. PMID- 8420786 TI - Indium-111-labelled antimyosin antibody imaging in a patient with cardiac sarcoidosis. AB - The aetiology of cardiac dysfunction caused by sarcoid granulomatous inflammation may be difficult to clarify, and the potential of imaging methods is limited. We report on a patient who presented with acute biventricular decompensation. Pulmonary sarcoidosis was confirmed after hospitalization. Four weeks after the initiation of corticosteroid treatment, scintigraphy with indium-111-labelled antimyosin antibody Fab fragments (AMAB) revealed distinct activity accumulation in major parts of the left ventricular wall (heart-lung ratio: 1.6) 72 h following injection. There may be a role for AMAB scintigraphy in the early detection of cardiac sarcoidosis. PMID- 8420787 TI - Value of ellipsoid volume masking in myocardial tomography. AB - The circumferential profile technique, used to obtain a bull's eye image from short axis slices in myocardial perfusion tomography, requires isolation of the reconstructed volume from other neighbouring structures that may take up the radiopharmaceutical used. If this is not possible, some regional artefacts can be introduced in the polar map that do not represent the actual myocardial perfusion at the corresponding level. In this case report we describe a method for volume masking that permits the myocardium inside in ellipsoidal volume to be enclosed. This technique is compared with the spherical method used in a Sophy 20P system in a case with impaired perfusion of an inferolateral segment and an extracardiac "hot" structure located in the same area. The results obtained show the usefulness of our method in such cases. PMID- 8420788 TI - Novel regulation of 5-HT1C receptors: down-regulation induced both by 5-HT1C/2 receptor agonists and antagonists. AB - The 5-hydroxytryptamine1C (5-HT1C) receptor shares many features with the 5-HT2 receptor. To determine if the regulation of the sites is also similar we studied the effects of chronic treatment with drugs active at 5-HT1C/2 receptors on [3H]mesulergine-labelled 5-HT1C binding sites in spinal cord. The 5-HT receptor agonists 1-(3-chlorophenyl)piperazine (m-CPP) (-38%), 1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane (DOI) (-35%), quipazine (-27%) and m trifluoromethylphenylpiperazine (TFMPP) (-27%) significantly down-regulated spinal 5-HT1C sites with chronic injection compared to vehicle treatment. The 5 HT receptor antagonists methiothepin (-71%), mianserin (-24%), methysergide ( 21%), and cyproheptadine (-27%) also induced down-regulation, and ritanserin and metergoline further reduced [3H]mesulergine specific binding to undetectable levels. There were no significant changes in Kd to implicate presence of residual drug except for mianserin, methiothepin, and TFMPP. Pindolol and spiperone had no significant effects. In acute dose-response studies, injection of a single dose of DOI did not result in a significant change in any receptor parameters. The capacity of a drug to lower Bmax correlated significantly with its pKd (r = 0.84, P < 0.0007). This drug regulation pattern for 5-HT1C sites of down-regulation by both 5-HT1C/2 receptor agonists and antagonists is similar to that for 5-HT2 receptors and is consistent with the classification of 5-HT1C and 5-HT2 receptors in the same superfamily. PMID- 8420789 TI - Characterization of muscarinic receptors in rat kidney. AB - Muscarinic receptors in mammalian kidney seem to be involved in diuresis. In this study we give a detailed characterization of receptors in rat kidney. Specific binding of [3H](-)-quinuclidinylbenzilate ([3H]QNB) to membranes of rat kidney cortex was saturable and of high affinity. A dissociation constant of 0.063 +/- 0.003 nM and a receptor density of 1.46 +/- 0.07 pmol/g wet weight were obtained. The dissociation kinetics could be best described by assuming a mono-exponential function (k-1 = (0.52 +/- 0.1) x 10(-4) s-1). The binding of [3H]QNB reached a maximum in 60 min at 0.6 nM at 37 degrees C. Competition experiments with the enantiomers of benzetimide confirmed the muscarinic nature of the [3H]QNB binding sites. The inhibition constants of pirenzepine (0.23 +/- 0.02 microM), (+-) hexahydrosiladifenidol (0.040 +/- 0.002 microM), AF-DX 116 (1.45 +/- 0.07 microM), methoctramine (1.67 +/- 0.02 microM) and gallamine (78 +/- 3 microM) classified this receptor as an M3 receptor. Inhibition of [3H]QNB binding by the agonists methylfurtrethonium, arecoline, isoarecoline methiodide, arecaidine propargyl ester and McN-A-343 displayed monophasic inhibition curves. With (+/-) cis-2-methyl-4-dimethylaminomethyl-1,3- dioxolane methiodide in two out of four experiments a small (11%) population of high affinity agonist sites could be detected. The potassium sparing diuretic amiloride inhibited [3H]QNB binding (36 +/- 3 microM). Although in a way related to the amiloride binding site, the muscarinic receptors in rat kidney are unlikely to be the primary target of diuretic action of this drug. PMID- 8420790 TI - Effects of nitric oxide-containing compounds on increases in cytosolic ionized Ca2+ and on aggregation of human platelets. AB - The present study was undertaken to determine the modulatory effects of nitric oxide (NO)-releasing compounds on increases in cytosolic ionized calcium ([Ca2+]i) and on aggregation of gel-filtered human platelets induced via diverse agonists. We used various sydnonimines and organic nitrates as donors of NO. Gel filtered and fura-2-loaded platelets were stimulated with ADP (4-8 microM), collagen (2-10 micrograms/ml) or thrombin (0.02-0.05 IU/ml), respectively. Half maximal inhibiting effects of sydnonimines on agonist-evoked increases in [Ca2+]i were observed between 30 and 1000 nM, while half-maximal inhibiting effects of the compounds on aggregation were between 3 and 500 nM. The compound C 87-3754, which is the bioactive metabolite of pirsidomine, was a much stronger inhibitor of increases in [Ca2+]i than of platelet aggregation. This was due to an enhanced NO release from this compound exposed to ultraviolet light during Ca2+ measurement. The organic nitrates isosorbide 5-mono-nitrate and nicorandil inhibited both aggregation and increase of cytosolic ionized calcium in stimulated platelets at half-maximal concentrations of approximately 200 microM. The present results suggest that some of the effects of NO on platelets are independent of cytosolic ionized calcium. The results also suggest that some of the inhibitory effects of NO-releasing compounds correspond rather to the presence of the A forms (NO-containing intermediates) than to the presence of free NO. PMID- 8420792 TI - Expression of alternatively-spliced glutamate receptors in human hippocampus. AB - Rat glutamate receptors have been shown to be expressed as two developmentally regulated, alternatively spliced isoforms. We have investigated the expression of these isoforms of GluRA and GluRB in the human hippocampus. The expression pattern of the mRNAs coding for these subunits does not correspond to that in the rat hippocampus, both isoforms being preferentially expressed in the dentate gyrus and CA1 regions, with lower expression in CA3, with the exception of GluRB flop, where hybridization in CA3 is only lower than in dentate gyrus. Cloning of cDNA from human frontal cortex has also revealed that the two isoforms of human GluRB have virtual nucleotide sequence identity with the alternative exons in the rat, confirming the usefulness of oligonucleotides complementary to the rat cDNAs as probes for these receptor subunits in human neuropsychiatric disorders. PMID- 8420791 TI - Muscarinic M3 receptors mediate total inositol phosphates accumulation in murine HSDM1C1 fibrosarcoma cells. AB - Muscarinic receptors in murine fibrosarcoma HSDM1C1 cells were characterized using both radioligand binding and total inositol phosphates accumulation (IPs). Muscarinic agonists elicited a concentration-dependent enhancement of IPs accumulation with a maximum of 14-fold stimulation above basal level. The following potencies (-log EC50) were observed for the full agonists: (+)-cis dioxolane 5.4, oxotremorine-M 5.3, (+)-muscarine 5.2 and carbachol 5.0. Bethanechol (4.1) and arecoline (5.0) were partial agonists, evoking 43 and 55%, respectively of the maximum level of stimulation to (+)-cis-dioxolane, whereas pilocarpine and McN-A-343 were inactive as agonists (1 mumol/l-1 mmol/1). The apparent affinities for muscarinic antagonists (-log KB) estimated by Schild regression were: 4-DAMP (4-diphenylacetoxy-N-methylpiperidine methiodide) 9.2, dicyclomine 7.0, pirenzepine 6.9, (+/-)-p-F-HHSiD (para-fluoro-hexahydro siladifenidol) 7.0, AF-DX 116 6.2, methoctramine 5.7. In saturation binding studies using [3H]N-methylscopolamine a homogeneous population of sites was identified, with a density of 145 pmol/mg protein. In competition radioligand binding studies, the following apparent affinities (-log Ki) were observed: 4 DAMP 9.7, dicyclomine 8.3, (+/-)-p-F-HHSiD 7.6, AF-DX 116 6.8, methoctramine 6.6 and gallamine 6.8. In binding studies all antagonists studied recognized a single population of sites, as judged by the Hill coefficients from the displacement isotherms. These data are consistent with HSDM1C1 cells expressing an apparent homogeneous muscarinic M3 population that mediates a large level of total IPs accumulation. This clonal line may provide a useful model to further elucidate relationship between endogenous muscarinic M3 receptor stimulation and IPs accumulation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420793 TI - Novel 2-substituted cocaine analogs: binding properties at dopamine transport sites in rat striatum. AB - A novel scheme utilizing vinylcarbenoid precursors has been developed for the synthesis of novel tropane analogs of cocaine. Using this method, 15 analogs were prepared and tested for activity in binding to dopamine transporters in rat striatal membranes using [125I]RTI-55. In all the analogs, the aryl group at the 3 position was directly bound to the tropane ring (as in WIN 35,428), and methyl or ethyl ketone moieties were present at the 2 position instead of the typical ester group. The most potent analog was a 2-naphthyl derivative (IC50 value of 0.2 nM, vs. 170 nM for cocaine), while replacement of the aryl with either ethyl or cyclohexyl drastically reduced potency (to > 50 microM and 5 microM, respectively). PMID- 8420794 TI - Structure and sequence of an intronless gene for human casein kinase II-alpha subunit. AB - Using sixteen different primers based on the cDNA sequence of the human casein kinase II-alpha subunit, different fragments of this gene were amplified by PCR from human genomic DNA. The sizes of these fragments were identical to amplified cDNA, which suggests the existence of an intronless genomic gene. The amplification was carried out on whole blood genomic DNA from three different individuals. The total sequence of the amplified casein kinase II-alpha gene showed more than 99% homology to the cDNA. The gene contains a noninterrupted open reading frame, as expected for the homolog cDNA. Although the gene sequence is complete, four point mutations were found. Since there are no interruptions of the open reading frame, this intronless gene might be expressed. PMID- 8420795 TI - Lipid transfer proteins (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens. AB - Four homogeneous proteins (Cw18, Cw20, Cw21, Cw22) were isolated from etiolated barley leaves by extraction of the insoluble pellet from a Tris-HCl (pH 7.5) homogenate with 1.5 M LiCl and fractionation by reverse-phase high-performance liquid chromatography. All 4 proteins inhibited growth of the pathogen Clavibacter michiganensis subsp. sepedonicus (EC50s = 1-3 x 10(-7) M) and had closely related N-terminal amino acid sequences. The complete amino acid sequences of proteins Cw18 and Cw21 were determined and found to be homologous to previously described, non-specific lipid transfer proteins from plants (32-62% identical positions). The proteins also inhibited growth of the bacterial pathogen Pseudomonas solanacearum (EC50s = 3-6 x 10(-7) M) and the fungus Fusarium solani (EC50s = 3-20 x 10(-6) M). A homologous protein from maize leaves (Cw41) was purified in a similar manner and also found to have inhibitory properties. A synergistic effect against the fungus was observed when protein Cw21 was combined with thionins. A defense role for non-specific lipid transfer proteins from plants is proposed. PMID- 8420796 TI - Stabilization of xylanase by random mutagenesis. AB - Four heat-resistant mutants of xylanase (N56, N102, N104 and F1) were obtained by random mutagenesis. The mutant genes had the following amino acid changes: N56, Ser-26 to Trp, Gly-38 to Asp and Thr-126 to Ser; N102, Gly-38 to Asp; N104, Gly 38 to Ser and Arg-48 to Lys; F1, Ser-12 to Cys. Kinetic studies showed that N104 is stabilized by an increase in the activation enthalpy, while the other mutants are stabilized by a decrease in the activation entropy. PMID- 8420797 TI - Purification and sequencing of cytochrome b from potato reveals methionine cleavage of a mitochondrially encoded protein. AB - Several mitochondrial genes from a large number of different fungi, mammals and plants have been sequenced but little is known about the corresponding translation products. We have affinity purified cytochrome c reductase from potato mitochondria and isolated the mitochondrially encoded cytochrome b protein. Amino-terminal sequencing reveals that the polypeptide does not start with a methionine. Comparison of the amino acid sequence with the recently published sequence of the gene encoding the cytochrome b apoprotein suggests that the N-formylmethionine is removed. This result provides the first evidence for the presence of a deformylase and a methionine aminopeptidase in mitochondria. PMID- 8420798 TI - cDNA cloning and expression of an Arabidopsis GTP-binding protein of the ARF family. AB - A cDNA clone encoding a small GTP-binding protein, the ADP-ribosylation factor (ARF) was isolated from a cDNA library of Arabidopsis thaliana cultured cells. The predicted amino acid sequence was highly homologous to the known yeast, bovine and human ARF sequences. Southern analysis of Arabidopsis genomic DNA suggested the existence of at least two copies of ARF genes. The level of ARF mRNA was found to be nearly constant during all cell growth stages in suspension cultures. PMID- 8420799 TI - Identification and characterization of an anti-tyrosine kinase factor in cystic gliomas. AB - In view of the frequent activation of the epidermal growth factor receptor (EGF R) in gliomas and autocrine hypothesis, we searched for 'EGF-like' factor(s) in cystic fluids (CFs) associated with gliomas. Membranes of A431 cells, which overexpress EGF-R, were used to explore such activity in 20 CFs. In all cases CFs induced inhibition of EGF-R phosphorylation. Biochemical analysis revealed an anti-tyrosine kinase activity which was identified as a 18 kDa proteic factor. Effectiveness at high dilution and anti-proliferative effect on living cells in culture suggest that this factor may be involved in the negative regulation of glial oncogenesis. PMID- 8420800 TI - Effect of modification of carbohydrate component on properties of glucoamylase. AB - In this study, we investigated enzymatic deglycosylation of glucoamylase from Aspergillus awamori X 100/D27, a glycoprotein which has two N-linked and about forty short mannose-bearing O-linked sugars per molecule. O-Linked sugars were modified by treatment with alpha-mannosidase and N-linked sugars were removed using endo-beta-N-acetylglucosaminidase F. Analysis of conformational changes following deglycosylation suggests that O-linked sugars essentially contribute to the stabilization of glucoamylase domains. Modification of the carbohydrate component by adding 1-deoxymannojirimycin to the culture medium induced inhibition of alpha-mannosidases involved in the processing, leading to a more complete glycosylation and, consequently, to a higher stability of the enzyme. PMID- 8420801 TI - c-fos gene expression in cell revertants from a transformed to a pseudonormal phenotype. AB - c-fos gene expression in two types of mouse sarcoma cells of spontaneous origin and in revertants to pseudonormal phenotype has been investigated. In the latter cells the content of c-fos mRNA is similar to that in normal fibroblasts. Activity of transcription factors interacting with the regulatory elements, SRE, DSE and TRE, in the c-fos promoter do not correlate with the c-fos mRNA concentration. However, experiments with cells transformed with the indicator plasmid, fos-CAT, showed that the 600 bp c-fos promoter region provides the chloramphenicol acetyltransferase activity correlating with c-fos mRNA expression in cell revertants to a pseudonormal phenotype. PMID- 8420802 TI - Cloning and characterization of the elongation factor EF-1 beta homologue of Saccharomyces cerevisiae. EF-1 beta is essential for growth. AB - A Saccharomyces cerevisiae cDNA homologue of the elongation factor EF-1 beta was found among the clones obtained by immunoscreening of a yeast cDNA expression library with an antibody against calmodulin affinity-purified proteins. The cDNA encoded a protein of 206 amino acids which was highly homologous (about 70% homology) with Artemia salina and human EF-1 beta. A protein with an apparent molecular mass of 33,000, significantly larger than that expected from the gene, was identified by Western blotting. Gene disruption experiments indicated that EF 1 beta is essential for growth. PMID- 8420803 TI - Molecular weight-determination of biosynthetically modified monomeric and oligomeric muropeptides from Escherichia coli by plasma desorption-mass spectrometry. AB - The presence of certain D-amino acids in the growth media of Escherichia coli results in the accumulation of 2 major and 3-5 minor new muropeptides in the murein sacculus. Preliminary data suggested that the major muropeptides correspond to a monomer and a cross-linked dimer with one residue of D-amino acid per molecule. We have analyzed several D-amino acid-modified muropeptides by plasma desorption-mass spectrometry. Our results confirmed that the general structures of the major modified muropeptides are: GlucNAc-MurNAc-L-Ala-D-Glu-m A2pm-D-X, and GlucNAc-MurNAc-L-Ala-D-Glu-m-A2pm-D-Ala; GlucNAc-MurNAc-L-Ala-D-Glu m-A2pm-D-X, being X a residue of the D-amino acid. These results corroborate the usefulness of this technique for the structural analysis of muropeptides. PMID- 8420804 TI - Conformational changes in subdomain-2 of G-actin upon polymerization into F-actin and upon binding myosin subfragment-1. AB - The susceptibility of subdomain-2 of actin to different proteases has been examined, for G-actin, F-actin, G-actin-S1(A2) and F-actin-S1(A2) complexes on a comparative basis. The sites of subtilisin, alpha-chymotrypsin and trypsin attack, exposed on G-actin, are protected in F-actin, F-actin-S1(A2) as well as in the G-actin-S1(A2) complex. In contrast, a new cleavage site (Arg39-His40) for ArgC protease, which is protected in G-actin, is exposed in G-actin-S1(A2) as well as in F-actin and F-actin-S1(A2). These results are consistent with the previously proposed structural analogy between the ternary (G-actin)2S1 and the F actin-S1 complexes, and provide information on the mechanism of S1-induced polymerization of G-actin. PMID- 8420805 TI - Characterization of metabotropic glutamate receptors coupled to a pertussis toxin sensitive G-protein in bovine brain coated vesicles. AB - Glutamate metabotropic receptors (mGluRs) in bovine brain coated vesicles have been characterized by pharmacological and kinetic binding experiments. Saturation experiments revealed a single binding site with a Kd = 607.9 +/- 78.5 nM and a Bmax = 6.45 +/- 0.88 pmol/mg protein. The specific binding of L-[3H]glutamate to mGluRs is regulated by guanine nucleotides. Guanosine-5'-triphosphate (GTP; 100 microM) shifts the agonist competition curves to the right, increasing the IC50 values. Pertussis toxin treatment produces a pharmacological binding profile for quisqualate similar to that obtained in the presence of 100 microM GTP. These results indicate the presence of metabotropic glutamate receptors in coated vesicles and its coupling to a pertussis toxin sensitive G-protein. PMID- 8420806 TI - Attenuated fibrinolysis and accelerated atherogenesis in type II diabetic patients. AB - Patients with hyperinsulinemia, defined by increased concentrations of IRI in plasma, experience increased cardiovascular mortality. In type II diabetic patients, the increase in IRI may reflect, in part, not only insulin but also proinsulinemia as a result of impaired conversion of proinsulin to insulin by pancreatic beta-cells. High IRI is accompanied by attenuation of endogenous fibrinolytic activity and increased plasma PAI-1, the primary physiological inhibitor of t-PA. Concordant increases of plasma PAI-1 and plasma IRI appear to reflect direct effects of insulin and proinsulin on the synthesis and secretion of PAI-1 by endothelial and liver cells as judged from results of studies in vitro. Because attenuated fibrinolysis may predispose to thrombosis, the increased exposure of luminal surfaces of vessels to atherogenic, clot-associated mitogens and chemoattractants may activate macrophages and potentiate proliferation of vascular smooth muscle cells. Accordingly, increased concentrations of plasma IRI may contribute to macrovascular disease in diabetic patients by impairing endogenous fibrinolysis. PMID- 8420807 TI - Distribution of type II diabetes in nuclear families. AB - Type II diabetes has a substantial genetic component, but the mode of inheritance and the molecular basis of this inheritance are uncertain. This study documents the familial distribution of the disease in the parents and siblings of a consecutive series of type II diabetic subjects. We studied 66 first-degree relatives of 20 white subjects with type II diabetes and both parents alive. They were tested with a continuous infusion of glucose (5 mg.kg IBW-1.min-1) (n = 49) or FPG and hemoglobin A1c (n = 17). Seven probands had neither parent affected with diabetes or IGT, 10 had one parent affected (6 with diabetes and 4 with IGT), and 3 had both parents affected. The probands with affected and those with unaffected parents were phenotypically similar. These findings indicate that a sizable subgroup of type II diabetic subjects may have neither parent affected with a demonstrable abnormality of glucose tolerance. The assumption of autosomal dominance with complete penetrance is not supported, although it remains possible that a dominant gene of low penetrance may play a role in some pedigrees. Polygenic inheritance would appear likely, and genetic heterogeneity may occur. The inheritance of diabetic traits from phenotypically normal parents needs to be considered in the analysis of genetic linkage with type II diabetes. PMID- 8420808 TI - Role for protein kinase C in the mediation of increased fibronectin accumulation by mesangial cells grown in high-glucose medium. AB - The fibronectin content of RMC cultures grown for 8-14 days in medium containing 30 mM (540 mg/dl) D-glucose was increased 30-60% relative to that of control cells cultured in 10 mM (180 mg/dl) glucose. Addition of equiosmolar concentrations (20 mM, 360 mg/dl) of L-glucose, 3-O-methylglucose, or mannitol to 10 mM glucose media did not alter fibronectin accumulation compared with values observed in 10 mM glucose alone. The basal phosphorylation of the 80,000-M(r) MARKS protein, which is a substrate for PKC, and the phosphorylation induced by acute (15-min) exposure of cells to 15% FCS or to 0.1 microM (50 ng/ml) PDBu were all increased in cells grown in 30 mM compared with 10 mM glucose. By contrast, phosphorylation of the 80,000-M(r) protein in response to a maximal concentration of PDBu (1 microM, 500 ng/ml) was not different in cells grown in 30 mM compared with 10 mM glucose. The acute increases in phosphorylation of the 80,000-M(r) protein were blocked by the PKC inhibitor calphostin C. Chronic (7-day) exposure of mesangial cells grown in 10 mM glucose to 0.1 microM of the PKC agonist PDBu also resulted in a sustained 40% increase in 80,000-M(r) phosphorylation and a 20 30% increase in fibronectin accumulation. As assessed by [35S]methionine incorporation, mesangial cell fibronectin synthesis was increased by exposure to PDBu, a finding consistent with earlier observations with 30 mM glucose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420809 TI - Effect of hypoxia on insulin secretion by isolated rat and canine islets of Langerhans. AB - The effect of pO2s reduced below physiological levels on GSIR by isolated islets of Langerhans was investigated with a microperifusion apparatus that provided control of pO2 and rapid dynamic response. Second-phase insulin secretion was reduced substantially by hypoxia. The response to lower pO2 was rapid and reversible. Although the steady, normoxic (pO2 = 142 mmHg) second-phase secretion rate varied widely from one islet preparation to another, the ratio of Sx to S142 for each preparation could be represented by a single curve that exhibited a continuous reduction with decreasing pO2. For rat islets perifused 1 day after isolation, the secretion rate was nearly 100% of the normoxic value at a pO2 of 60 mmHg, 50% at 27 mmHg (P50, the pO2 at which the S142 is reduced by 50%), and approximately 2% at 5 mmHg. Oxygen sensitivity of second-phase secretion rate declined after 1 wk of in vitro culture: P50 was 13 mmHg after 1 wk and remained at 10 mmHg after 2-5 wk of culture. Canine islets exhibited a P50 of 16 mmHg after 1 wk of culture. The reduction in insulin secretion is thought to be associated with the existence of pO2 gradients outside and inside the isolated islets, resulting in exposure of islet cells to low pO2 levels that decrease radially from the periphery to the core. We hypothesize that the effect of low pO2 on S is manifested through depletion of the energy stores of the beta-cells. The effect of hypoxia on S may be an important factor in some in vitro secretion studies and may play a critical role in the effectiveness of transplanted islets before their revascularization and of immunoisolated islet implantation devices. PMID- 8420810 TI - Increase in simultaneous coexpression of naive and memory lymphocyte markers at diagnosis of IDDM. AB - The monoclonal antibodies 2H4 (anti-CD45RA) and UCHL1 (anti-CD45RO) were used to subdivide the CD4 and CD8 T-cell subsets into naive and memory cells. The peripheral blood lymphocytes of 34 patients with recent-onset IDDM, 21 patients with long-standing IDDM, and healthy control subjects of similar age and sex were analyzed by a three-color immunofluorescence technique. CD4 and CD8 lymphocytes expressed the CD45 isoforms alone (CD45RA+ or CD45RO+) or in combination CD45RA+RO+). Simultaneous coexpression of both CD45RA and CD45RO (CD45RA+RO+) on CD4 and CD8 lymphocytes in patients with recent-onset IDDM was higher than in control subjects (P < 0.001). The proportion of CD4 lymphocytes expressing CD45RA alone (CD45RA+RO-) was similar in these groups, but the percentage of CD8 lymphocytes that were CD45RA+RO- was significantly higher in the patients with recent-onset IDDM (P < 0.05). The result of these changes is a significant increase in expression of naive phenotypes (CD45RA+ and CD45RA+RO+) on CD4 and CD8 lymphocytes in recent-onset IDDM (P < 0.005 and P < 0.0001). In long-standing IDDM, total CD45RA+ expression on CD4 and CD8 lymphocytes was reduced compared with control subjects (P < 0.05) as a result of a tendency of CD45RA+RO- and CD45RA+RO+ subsets to be lower. This increase in total naive (CD45RA+) lymphocytes and in coexpression of naive (CD45RA) and memory (CD45RO) markers on CD4 and CD8 lymphocytes subsets in patients with recent-onset IDDM suggests that abnormal regulation of T-cell activation and maturation is important in the pathogenesis of the disease. PMID- 8420811 TI - Endothelium-dependent vascular relaxation in patients with type I diabetes. AB - The endothelium plays an important role in the regulation of vascular tone. Although animal data show evidence for an impaired endothelium-dependent vasodilation in diabetes, human in vivo data are scarce. We investigated 11 type I diabetic patients and 11 matched healthy control subjects. The diabetic patients were selected on their relatively poor metabolic regulation (HbA1c > 8.5%), but none showed signs of microvascular complications. In all subjects, we recorded the forearm vasodilator responses to three different stimuli: 1) 5 min of forearm ischemia to obtain a maximal vasodilator response; 2) infusion of MCh into the brachial artery (dosages: 0.03-0.3-1.0 micrograms.min-1.100 ml-1 forearm volume) to evaluate endothelium-dependent vasodilation; and 3) intra-arterial infusion of SNP (dosages: 0.06-0.2-0.6 micrograms.min-1.100 ml-1) to evaluate endothelium-independent vasodilation. The diabetic patients had their usual subcutaneous insulin dose and breakfast 90 min before the start of the test. Baseline levels of BP and FBF were similar in both groups. The PORH response was similar in both groups, with a percentage fall in FVR of 92 +/- 1% in diabetic patients and 94 +/- 1% in control subjects. In the control subjects, MCh infusions exerted a dose-dependent vasodilator response with a maximal fall in the FVR of 90 +/- 2%. The highest dose of SNP induced a fall in FVR of 81 +/- 6% in this group. In diabetic patients, the percentage decrements in FVR during the several dosages of MCh and SNP were similar when compared with the control group. We conclude that chronic hyperglycemia, as occurred in our patients with uncomplicated diabetes mellitus, does not impair endothelium-dependent vasodilation in vivo. PMID- 8420812 TI - Incidence of NIDDM and impaired glucose tolerance in hypertensive subjects. The San Antonio Heart Study. AB - Hypertension often occurs in association with NIDDM and IGT. We examined the association of hypertension at baseline to the 8-yr incidence of NIDDM and IGT in 1471 subjects who participated in the San Antonio Heart Study. Subjects who were hypertensive at baseline had a higher incidence of NIDDM (8.9 vs. 4.9%, P = 0.041) and IGT (25.2 vs. 10.0%, P < 0.001) than subjects who were normotensive at baseline. After adjusting for age, sex, ethnicity, obesity, body fat distribution, fasting glucose, and insulin, this excess was eliminated for NIDDM, but not for IGT. Specifically, the adjusted OR for NIDDM in hypertensive versus normotensive patients was 0.73 (95% Cl 0.34-1.58), whereas the adjusted OR for IGT was 1.87 (95% Cl 1.08-3.22). The excess risk of NIDDM in hypertensive patients can be explained by their greater age, obesity, more unfavorable body fat distribution, and hyperinsulinemia, whereas the excess risk of IGT is independent of these factors. PMID- 8420813 TI - Normal proinsulin responses to glucose in mild type II subjects with subnormal insulin response. AB - IPI, 32-33 SPI, and insulin were measured by specific assays and related to plasma glucose and BMI in diet-treated type II diabetic subjects (FPG 7.3 +/- 1.8 mM) and nondiabetic control subjects, both fasting and during a 12-mM hyperglycemic clamp. In both groups, BMI correlated with fasting plasma insulin (rs = 0.76, P < 0.001 and 0.50, P < 0.01, respectively) and IPI (rs = 0.49, P = 0.03 and rs = 0.69, P < 0.001, respectively). Accounting for obesity, fasting plasma insulin was subnormal in diabetic subjects (58% of control group, 1 SD range, 49-68%), but did not correlate with FPG. In contrast, fasting plasma IPI correlated with FPG in the diabetic patients (rs = 0.47, P < 0.05). In all subjects, 64% of the variance in plasma IPI was explained by BMI and FPG. Fasting 32-33 SPI was similar in the two groups. In response to a hyperglycemic clamp, the diabetic subjects had subnormal insulin concentrations (geometric means 71 and 214 pM, P < 0.001), but normal IPI concentrations (11.6 and 14.2 pM, respectively). Reduction of 32-33 SPI concentrations in diabetic subjects was intermediate (7.3 and 13.2 pM, P < 0.05). In diabetic subjects both fasting and clamp responses were subnormal for insulin but apparently normal for IPI. The major defect in pancreatic function is an impaired insulin response to glucose, and this, rather than an increase in proinsulin secretion, gives rise to the relative increase in proinsulin. PMID- 8420814 TI - Effect of high glucose on type IV collagen production by cultured glomerular epithelial, endothelial, and mesangial cells. AB - Immunochemical and metabolic radiolabeling procedures revealed that homogeneous cultures of calf glomerular epithelial, endothelial, and mesangial cells actively synthesize type IV collagen (primarily as alpha 1 (IV)3) which is secreted into the medium and incorporated into the extracellular matrix. Exposure of confluent cultures of the three cell types to a high glucose concentration (30 mM) for 60 h resulted in a pronounced increase (two- to threefold) in type IV collagen production over that observed at a physiological level (5 mM) of this sugar, as determined by either immunoblotting or fluorography of electrophoretically separated media or cell-matrix components. The elevated glucose did not bring about a change in the rate of cell proliferation or fibronectin production. Moreover, studies with mannitol indicated that the stimulation of type IV collagen synthesis was not a function of hyperosmolarity. In contrast to the glomerular cells, glucose-induced enhancement of formation of this collagen was not observed in 3T3 cells despite a substantial acceleration in the consumption of this sugar. Time studies indicated that the response of the glomerular cells to high glucose occurs over an extended period (maximal at approximately 78 h) and, furthermore, that the stimulatory effect on type IV collagen production is only slowly reversed after restoration of the glucose to a normal level. We believe that these findings are relevant to an understanding of the sequence of events that lead to the development of diabetic glomerular lesions. PMID- 8420815 TI - Picotamide, a dual TXB synthetase inhibitor and TXB receptor antagonist, reduces exercise-induced albuminuria in microalbuminuric patients with NIDDM. AB - We investigated the short-term effect of the TXB inhibitor picotamide on albuminuria induced by exercise in 15 microalbuminuric (i.e., with UAE at rest between 20 and 200 micrograms/min) type II diabetic patients (12 men and 3 women, age 56 +/- 2, BMI 28 +/- 1 kg/m2) and in six normal age-matched control subjects. The diabetic subjects performed five submaximal exercise tests (90% of theoretical heart rate) on a cycle ergometer: the first two under basal conditions; the third and fifth after subjects had received picotamide (900 mg/day) or placebo (3 tablets/day) for 10 days; the fourth exercise always was performed after 10 days of wash-out. Control subjects performed two exercises: the first in baseline conditions and the second after 10 days of picotamide administration (900 mg/day). When diabetic patients were untreated, a significant (P < 0.05) increase in UAE with respect to baseline levels was observed immediately after and 1 h after the exercise test. After picotamide administration, UAE significantly decreased (P < 0.05) immediately after and 1 h after exercise, as compared with diabetic patients given a placebo. In normal subjects, exercise was followed by a slight increase in UAE, which was not significantly affected by picotamide administration. Our results show that short term administration of picotamide is associated with a reduction in UAE after exercise in type II diabetes patients with microalbuminuria while at rest. Picotamide, a TXB synthetase and receptor inhibitor, may decrease exercise induced albuminuria in diabetic patients through a reduction in circulating TXB levels and inhibition of TXB action, which in turn may act by lowering glomerular capillary hydraulic pressure. PMID- 8420816 TI - Fibronectin-induced increase in mesangial cell prostaglandin release. Effect of hyperglycemia and PKC inhibition. AB - Glomerular accumulation of extracellular matrix in diabetes is a potential regulator of mesangial cell-matrix interactions through transmembrane matrix receptors. We now provide evidence that PG production from rat glomerular mesangial cells is increased by Fn. An increase in PG (measured as PGE) was demonstrated in mesangial cell-enriched glomerular cores after 1-h exposure (149 +/- 8% of timed control) and was sustained over a 24-h period (214 +/- 7%). Increased PG production followed exposure to a chymotryptic fragment (120,000 M(r)) of Fn and occurred concomitant with an increase in particulate PKC activity. A tetrapeptide (Arg-Gly-Asp-Ser) with the Arg-Gly-Asp sequence, contained in Fn and the chymotryptic fragment and recognized by specific membrane receptors (integrin matrix-binding proteins), also raised PG levels. As has been shown previously, exposure to high glucose concentration can increase mesangial cell PGE production (from 677 +/- 61 pg.mg protein-1.2 h-1 at 5.6 mM glucose to 1561 +/- 132 pg.mg protein-1.2 h-1 at 50 mM glucose, P < 0.001). The response to the chymotryptic fragment of Fn also was enhanced by concurrent exposure to high glucose concentration (from 2560 +/- 199 pg.mg protein-1.2 h-1 at 5.6 mM glucose to 4672 +/- 358 pg.mg protein-1.2 h at 50 mM glucose, P < 0.001). Coincubation with H-7, an inhibitor of PKC, abolished the PG response to glucose and the chymotryptic fragment. Involvement of PKC was supported further by abrogation of the effect of chymotryptic fragment in mesangial cells cultured for a prior prolonged period with phorbol ester.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420817 TI - Glucose modulates glucose transporter affinity, glucokinase activity, and secretory response in rat pancreatic beta-cells. AB - Pancreatic islets were cultured for 24 h in medium containing either low (1.4), normal (5.5), or high (16.7 mM) glucose, and then insulin secretion was measured at the end of 1 h incubation at 37 degrees C. Insulin release in the absence of glucose was 64 +/- 20, 152 +/- 11, and 284 +/- 30 pg.islet-1.h-1 (mean +/- SE, n = 6, G1.4 and G16.7 vs. G.5.5, P < 0.05) and the response to 22 mM glucose stimulation was 640 +/- 136, 2460 +/- 276, and 1890 +/- 172 pg.islet-1.h-1, respectively (n = 6, G1.4 vs. G5.5, P < 0.01, G16.7 vs. G5.5, P = 0.065). The 50% maximal response of insulin secretion (increment over baseline) was reached at an average glucose concentration of 9.9 +/- 0.7 mM in islets preexposed to G5.5, and at glucose 13.3 +/- 0.9 and 4.8 +/- 0.4 mM (P < 0.05 in respect to G5.5) in islets preexposed to G1.4 and G16.7, respectively. To investigate the molecular mechanism responsible for this altered glucose sensitivity, we measured, in parallel experiments, the kinetic characteristics of glucose transport, glucokinase, and glucose utilization. Glucose transport was measured by evaluating 3-O-methylglucose uptake. The apparent Km of the low-affinity transporter (GLUT2) was 16.6 +/- 2.4 mM in isolated pancreatic cells cultured at 5.5 mM glucose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420818 TI - Increased proinsulin/insulin ratio in pancreas extracts of hyperglycemic rats. AB - The plasma ratio of proinsulin/insulin is raised in people with NIDDM. A relative hypersecretion of proinsulin is thought to be the cause, because pancreas extracts from diabetic rats have a raised proinsulin/insulin ratio. We tested the hypothesis that the pancreatic proinsulin/insulin mismatch results from hyperglycemia-induced beta-cell degranulation. Normal rats made hyperglycemic with 48-h glucose infusions had a raised pancreatic percentage of proinsulin. In contrast, rats infused with enough glucose to induce compensatory hyperinsulinemia without changing the plasma glucose level had a normal percentage of proinsulin. The raised percentage of proinsulin in the hyperglycemic rats reflected a reduction in pancreatic insulin content. Administering an inhibitor of insulin release, diazoxide, to hyperglycemic rats blocked the fall in pancreatic insulin content and prevented the rise in the percentage of proinsulin. Normal rats infused with tolbutamide for 3 days and enough glucose to maintain euglycemia had a 50% reduction in pancreatic insulin content. The beta-cell degranulation from this nonhyperglycemic mechanism resulted in a raised pancreatic percentage of proinsulin. In summary, chronic hyperglycemia causes beta-cell degranulation primarily because of hyperstimulated insulin release. The net result is a rise in the ratio of immature (proinsulin rich) to mature (insulin-rich) granules, which is reflected as an increased relative proportion of proinsulin. Mobilization of these proinsulin-enriched granules may explain the relative hypersecretion of proinsulin that occurs with diabetes. PMID- 8420819 TI - Effects of ethanol on carbohydrate metabolism in the elderly. AB - We have previously reported that in young men, ethanol caused acute insulin resistance, but compensatory insulin secretion prevented deterioration of glucose tolerance (1). In this study, we tested the hypothesis that elderly men, because of their pre-existing insulin resistance and compromised insulin secretory capacity, may experience worsening of their glucose tolerance after ethanol. Nine elderly men (65.7 +/- 0.8 yr, BMI 25.8 +/- 1.4 kg/m2) received ethanol (13 mmol/kg for 30 min i.v.) or saline followed 30 min later by i.v. glucose (2.8 mmol/kg for 5 min). To determine the mechanism of the ethanol effect, six of the men underwent euglycemic-hyperinsulinemic (approximately 350 pM) clamping with simultaneous infusion of ethanol or saline. Muscle biopsies were obtained before and 1 and 4 h after insulin infusion. In all nine men, glucose concentrations after i.v. glucose were higher after ethanol than after saline, whereas insulin was the same and glucose tolerance decreased by 23% (Kg 2.41 +/- 0.2 vs. 1.86 +/- 0.1%/min, P < 0.01). Ethanol reduced insulin-stimulated glucose uptake from 40.6 +/- 3.1 to 25.6 +/- 1.9 mumol.kg-1.min-1 (-37%, P < 0.05), glucose oxidation from 11.7 +/- 1.1 to 7.0 +/- 0.7 mumol.kg-1.min-1 (-33%, P < 0.01), and glucose storage from 28.7 +/- 2.4 to 18.6 +/- 1.7 mumol.kg-1.min-1 (-35%, P < 0.01). Ethanol increased muscle lactate concentration from 0.49 +/- 0.14 to 1.99 +/- 0.99 mumol/mg protein (P < 0.05), but had no effects on muscle concentration of free glucose, G-6-P, and citrate concentrations, nor did it affect muscle GS activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420820 TI - Interaction of interleukin-1 with islet beta-cells. Distinction between indirect, aspecific cytotoxicity and direct, specific functional suppression. AB - A 5-day culture of adult rat islets with human recombinant IL-1 beta (3 U/ml) resulted in the death of most alpha-cells and 50% of beta-cells. The IL-1- exposed islet tissue contained--in addition to poorly granulated beta-cells- patches of outgrowing monolayers and dispersed activated macrophages. In purified alpha- and beta-cell preparations, no cytodestructive effects of IL-1 (as high as 30 U/ml) were noticed, indicating that the cytokine is in itself not a beta-cell- selective killer. Pure beta-cells were, on the other hand, more sensitive (from 0.3 U/ml on) than intact islets to an IL-1--induced suppression of hormone synthesis. This inhibitory action was reversible and affected predominantly the production of insulin, leading to degranulated cells with modified shape and attachment. Further studies with IL-1 should take into account that isolated islet preparations do not allow distinction between its irreversible, indirect, and aspecific beta-cell toxicity and its reversible, direct, and specific suppression of beta-cell functions. It is not yet known whether IL-1--suppressed beta-cells exhibit an altered sensitivity to beta-cell--toxic conditions. PMID- 8420821 TI - Vascular thrombosis in type II diabetes mellitus. PMID- 8420822 TI - The role of histocompatibility antigens in transplantation of isolated islets of Langerhans in the rat. AB - In a model of congenic and intra-MHC recombinant rat strains, the differential role of various histocompatibility antigens in renal subcapsular transplantation of purified islets of Langerhans is evaluated. Class I MHC antigens of the RT1.A region, expressed on the endocrine cells of the islets themselves, do not induce graft rejection on their own. MHC class I antigens as encoded by the RT1.C region do not induce rejection either. MHC class II antigens as encoded by the RT1.B/D region are not expressed on the endocrine pancreas, not even during rejection. Although interstitial dendritic cells situated within the islets express these antigens, an isolated RT1.B/D incompatibility of islets is associated with prolonged survival in contrast to rapid rejection of fully MHC-mismatched grafts. Unlike other organs, islets matched for all MHC antigens, but incompatible at minor histocompatibility antigens, undergo rejection early after transplantation. PMID- 8420823 TI - Insulin secretory function in relation to transplanted islet mass in STZ-induced diabetic rats. AB - In vivo insulin secretion was quantified as the AIRg or AIRa in islet transplanted rats. Male Wistar-Furth rats previously made diabetic by STZ administration (55 mg/kg) were transplanted with 500, 1000, 2000, or 3000 islets infused into the portal vein (n = 12-14 per group) and were compared with sham treated controls (CN, n = 16). At 4-5 wk posttransplantation, no significant differences were noted in the FPG or fasting plasma insulin of the experimental groups (P > 0.05). Body weight, however, was 10% less (P < 0.05) in rats receiving 500 islets than in controls, indicating an effect of beta-cell deficiency on growth rates. To determine the relationship between islet mass and insulin secretion, we measured AIRg after a 0.3 g/kg glucose bolus in fasted conscious animals. A significant correlation was observed between the AIRg and islet number (r = 0.61, P = 0.0001), and both 500- and 1000-islet groups could be differentiated from controls by ANOVA (500: 8%; 1000: 12% of controls; P < 0.05). During a glycemic potentiation protocol, AIRa was measured at basal and elevated blood glucose (approximately 16 mM). At neither basal nor elevated blood glucose was AIRa correlated with islet number (basal r = 0.0622, P = 0.7834; elevated r = 0.3133, P = 0.1667). None of the groups could be differentiated by ANOVA (elevated 500: 37%; 1000, 68% of controls; P > 0.05). Although this study illustrates that AIRa may be better preserved in islet-transplanted rats, AIRg is the better correlate of islet number.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420824 TI - New T-cell receptor gamma haplotypes in wild mice and evidence for limited Tcrg-V gene polymorphism. AB - Tcrg gene polymorphism was investigated by Southern blot analysis on a panel of laboratory and wild mouse strains using a set of probes which identify all known Tcrg-V and -C genes. Only three haplotypes are found in laboratory mice: gA, gB, and gC which are represented by BALB/c, AKR, and DBA/2 prototypes respectively. gA and gC haplotypes are the most frequent among laboratory mice whereas gB is poorly represented. Seven new haplotypes are described among 23 wild mice corresponding to four Mus musculus subspecies (Mus mus domesticus, castaneus, musculus, and molossinus). However, only a few new alleles of individual genes are observed. Tcrg-V genes located at the 5' end of the Tcrg locus (V7 and V4) appear to be nonpolymorphic whereas two Tcrg-V3, -V5, -V6, -C4 and three Tcrg-V1, -V2, -C1, -C2, and -C3 specific restriction fragment length polymorphisms are detected. These results indicate a relatively high degree of conservation of Tcrg genes as compared to other members of the immunoglogulin (Ig) gene family and might be related to the specifity and function of gamma delta T cells. Several of the new haplotypes described here result from point mutations in noncoding Tcrg-V or -C gene-flanking regions. Recombinations may have also participated in the evolution of the Tcrg locus. Finally, these new Tcrg haplotypes are unequally distributed among the four M. m. subspecies and support the idea that the gA and gC haplotypes found in laboratory mice are inherited from M. m. domesticus whereas gB might originate from asian subspecies (castaneus, musculus or molossinus). PMID- 8420825 TI - Strong genetic association between HLA-DR3 and a polymorphic variation in the regulatory region of the HSP70-1 gene. AB - A three-allele polymorphic system is detectable by direct electrophoretic analysis of the amplified 5' untranslated und 5' flanking regions of the HLA linked HSP70-1 gene. Single nucleotide differences at two sites, i.e., -110 and +120, are responsible for changes in the bending pattern and, consequently, in the electrophoretic mobility of the three alleles. The presence of the sequence AAACCCC around position -110 is strongly associated with DR3. PMID- 8420826 TI - Isolation of a novel cDNA clone showing marked similarity to ME491/CD63 superfamily. AB - A novel cDNA clone, A15, was isolated by the differential screening of a cDNA library of an immature T cell line, HPB-ALL using radioactive cDNA probes from the mRNA of either HPB-ALL or peripheral blood lymphocytes. It hybridized to a single mRNA species of about 2.0 kilobases which is expressed in HPB-ALL cell line, but not in the PBL or a promyelocytic leukemia cell line, HL-60. The A15 gene codes for a protein of 244 amino acids which contains four potential transmembrane domains and four possible N-linked glycosylation sites. A computer aided comparison showed a marked similarity to several other membrane proteins: CD9, CD37, CD53, TAPA-1, Sm23, CO-029, and ME491/CD63. PMID- 8420827 TI - T-cell repertoire in a strain of transgenic C57BL/6 mice with the HLA-DRA gene on the X-chromosome. AB - We have established a strain of transgenic mice in which the HLA-DRA gene was integrated into the X-chromosome and the xenogeneic mixed isotype molecule, DR alpha E beta b, was expressed in a cell type-specific manner, although the transgenic DRA gene contained only 268 base pairs of the 5'-flanking region. The DR alpha E beta b molecules expressed in the transgenic mice functioned as major histocompatibility complex (MHC) class II to select T-cell repertoire, and to stimulate mixed lymphocyte reaction. In female transgenic mice homozygous for HLA DRA (DR alpha-B6-F-homo) and male transgenic mice (DR alpha-B6-M), DR alpha E beta b molecules were expressed in almost all of the MHC class II Ab-positive cells. In contrast, the expression of DR alpha E beta b molecules in female transgenic mice hemizygous for HLA-DRA (DR alpha-B6-F-hemi) was found only in part of the Ab positive cells, and the proportion of cells expressing the DR alpha E beta b molecules varied due to random inactivation of one of the X chromosomes. Clonal deletions of the T cells and mature thymocytes bearing Tcrb V5 and Tcrb-V11, which are eliminated from the peripheral repertoire in mice expressing self-superantigen and MHC class II E molecules, were incomplete in DR alpha-B6-F-hemi as compared with those in DR alpha-B6-F-homo, and were correlated with the proportion of DR alpha E beta b-positive spleen cells. These observations suggested that the number of bone marrow-derived cells expressing DR alpha E beta b molecules was critical for clonal deletions of Tcrb-V5+ and Tcrb V11+ T cells in the thymus. PMID- 8420828 TI - Molecular analysis of HLA-B39 subtypes. AB - Serological studies have suggested the presence of a new HLA-B39 subtype (B39.2) in the Japanese population. To identify the new HLA-B39 subtype and compare it with an other HLA-B39 subtype (B39.1), the genes encoding HLA-B39.1 (B*39013) and B39.2 (B*3902) have been cloned from Japanese. We have sequenced these genes and completed the sequence of HLA-B39.1 (B*39011) gene from a Caucasian that was partially sequenced. Comparison of the sequence data revealed that B*3902 and B*39013 differ by three nucleotide substitutions which result in a two amino acids change at residues 63 and 67, while one silent substitution at codon 312 is found between B*39011 and B*39013. These results suggest that B*3902 has evolved from B*39013 rather than B*39011. PMID- 8420829 TI - DST4: a new, and probably the last, functional DH gene in the BALB/c mouse. PMID- 8420830 TI - Sequence analysis of sheep T-cell receptor beta chains. PMID- 8420831 TI - Three in vivo promoter phenotypes in MHC class II deficient combined immunodeficiency. PMID- 8420832 TI - Sequencing and genetic analysis of a bovine DQA cDNA clone. PMID- 8420833 TI - HLA class I nucleotide sequences, 1992. AB - The HLA class I sequences included in this compilation are taken from publications listed in the papers: Nomenclature for factors of the HLA system, 1991 (Bodmer et al. 1992); Nomenclature for factors of the HLA system, 1990 (Bodmer et al. 1991); and Nomenclature for factors of the HLA system, 1989 (Bodmer et al. 1990). Due to the increased number of sequences we have only included sequences for exons 2, 3, and 4 in this compilation. Where discrepancies have arisen between reported sequences, the original authors have been contacted where possible, and necessary amendments to published sequences have been incorporated into this alignment. Future sequencing may identify errors in this list and we would welcome any evidence that helps to maintain the accuracy of this compilation. In the sequence alignments, identity between nucleotides is indicated by a hyphen (-). An unavailable sequence is indicated by a period (.). Gaps in the sequence are inserted to maintain the alignment between different alleles showing variation in amino acid number. PMID- 8420834 TI - HLA-DPB1 alleles in a population from south China. AB - Using the polymerase chain reaction (PCR) and hybridization with oligonucleotide probes, we analyzed the distribution of DPB1 alleles in 99 healthy unrelated individuals from the city of Guangzhou (Canton), South China. Twelve different DPB1 alleles were found in this panel. The most common allele was DPB1*0501 (62.6%). Other major alleles detected included DPB1*02 (DPB1*0201 and DPB1*0202), DPB1*1301, DPB1*0401, and a recently described allele, designated DPB1*2101. The hybridization pattern of DPB1*2101 showed that this allele shared sequences with DPB1*0301 and DPB1*0601 in the A and F hypervariable regions, while the C, D, and E regions were identical to those of DPB1*0202. DPB1*2101 was observed in 11% of the subjects tested. It was found to be in strong linkage disequilibrium with DRB1*1202. In family studies, segregation of the haplotype DRB1*1202, DRB3*0301, DQA1*0601, DQB1*0301, DPB1*2101 was observed. The second exon of DPB1*2101 was sequenced from codon 8 to codon 90 and the sequence, inferred from the pattern of hybridization, was confirmed. DPB1*0301, DPB1*0402, DPB1*0101, DPB1*1401, DPB1*1901, and another recently recognized allele, now designated DPB1*2401, were detected with low frequencies. DPB1*2401 had the same hybridization pattern as DPB1*0501 except for a probe that matches codons 85-90. In this region, DPB1*2401 encoded the amino acid sequence GPMTLQ instead of EAVTLQ as in DPB1*0501. PMID- 8420835 TI - Novel members and germline polymorphisms in the human T-cell receptor Vb6 family. AB - The human T-cell receptor (Tcr) Vb6 family has been scrutinized for polymorphisms, both in coding as well as in intronic sequences by polymerase chain reaction (PCR), subsequent multiple electroblot hybridizations, and sequence analysis. Multiplex PCR is an efficient means of screening for Tcr variability. Four novel loci could be distinguished and several new alleles are described including two pseudogenes. The Vb6 family is characterized by an intronic stretch of simple repetitive (gt)n sequences. These elements are hypervariable, especially in the Vb6.7 subfamily, where they are particularly long. The unexpected persistence of simple repetitive sequences in Tcr and major histocompatibility complex (MHC) class II genes over extended periods of the vertebrate evolutionary history can be interpreted in parallel terms in both gene families. PMID- 8420836 TI - Primate ABO glycosyltransferases: evidence for trans-species evolution. AB - The human ABO blood group system is controlled by alleles at a single locus on chromosome 9. The alleles encode glycosyltransferases, which add different sugar residues to the terminal part of the oligosaccharide core, thus generating the A or B antigens; an allele encoding enzymatically inactive protein is responsible for the blood group O. The A and B antigens are present not only in humans, but also in many other primate species and it has been proposed that the AB polymorphism was established long before these species diverged. Here we provide molecular evidence for the trans-species evolution of the AB polymorphism. Polymerase-chain reaction (PCR) amplification and sequencing has revealed that the critical substitutions differentiating the A and B genes occurred before the divergence of the lineages leading to humans, chimpanzees, gorillas, and orangutans. This polymorphism is therefore at least 13 million years old and is most likely maintained by selection. Comparison of the sequences derived from different species indicates that the difference in enzymatic activities between the A and B transferases is caused by two single nucleotide substitutions responsible for Leu-Met and Gly-Ala replacement at positions 265 and 267 in the polypeptide chains, respectively. PMID- 8420837 TI - Invariant components of the sheep T-cell antigen receptor: cloning of the CD3 epsilon and Tcr zeta chains. PMID- 8420838 TI - Complete characterization of the expressed immune response genes in Biozzi AB/H mice: structural and functional identity between AB/H and NOD A region molecules. PMID- 8420839 TI - Construction of a monoclonal antibody against Igh-7a. PMID- 8420840 TI - Identification of rat Tcrb-V8.2, 8.5, and 10 gene products by monoclonal antibodies. PMID- 8420841 TI - Structure of porcine MHC class II DRB genes. PMID- 8420842 TI - The nucleotide sequence of bovine MHC class II DQB and DRB genes. PMID- 8420843 TI - New Medicaid program promotes caring for elderly at home. PMID- 8420844 TI - Advice to transplant patients? Ask questions. AB - Organ transplantation has meant a miraculous prolongation of life for many Iowans. However, this young man's story--sent unsolicited to the editors of Iowa Medicine--illustrates what happens when a patient has unrealistic expectations of what life will be like after a transplant. PMID- 8420845 TI - Logic and consistency needed in long-term care. PMID- 8420846 TI - Suburethral sling--an old procedure revisited. PMID- 8420847 TI - Want to be a technician? PMID- 8420848 TI - Shared management. PMID- 8420849 TI - Nonautonomous patients. PMID- 8420850 TI - Children aren't second-class citizens. PMID- 8420851 TI - Caring for the elderly: a crisis on the horizon. AB - As baby boomers age, the proportion of elderly people will increase. Growing numbers of senior citizens coupled with shrinking budgets may force society to find alternatives to nursing homes, predicts a Des Moines geriatrician. PMID- 8420852 TI - Wolff-Parkinson-White syndrome and the use of radiofrequency catheter ablation. AB - OBJECTIVE: To describe Wolff-Parkinson-White (WPW) syndrome and the use of radiofrequency catheter ablation to prevent further recurrence of the tachycardias associated with this syndrome. The pathophysiology, electrocardiographic findings, treatment modalities for both short- and long-term therapy, as well as the treatment with radiofrequency catheter ablation are presented. PATHOPHYSIOLOGY: WPW is the most common form of the preexcitation syndromes. Ventricular myocardium is preexcited by use of an alternate conduction pathway or accessory pathway. Electrocardiographic characteristics show a short PR interval, presence of a delta wave, widened QRS complex, and ST-T wave changes. Most common tachyarrhythmias associated with this syndrome are orthodromic and antidromic reciprocating tachycardia and atrial fibrillation. OUTCOME: There is significant morbidity and mortality associated with WPW. Although rare, some patients' initial presentation may be ventricular fibrillation or sudden cardiac death. INTERVENTIONS: Short-term therapy will include vagal maneuvers, atrioventricular nodal blocking agents, or direct current cardioversion. Long-term therapy includes antiarrhythmic therapy, and surgical or catheter ablation. NURSING CONSIDERATIONS: Critical Care nurses play an important role in the diagnosis and treatment of the patient with WPW. Recognizing the characteristic signs on the 12-lead electrocardiogram, understanding the proper therapy for arrhythmias, and possessing knowledge of the syndrome to educate the patient are vital pieces of information necessary to care for the patient with WPW. PMID- 8420853 TI - Excimer laser angioplasty. AB - Excimer laser angioplasty is one of the newest procedures being used in interventional cardiology. Because of persistently high rates of restenosis with conventional balloon angioplasty (which may be attributed to the acceleration of atherosclerosis by remodeling and dilatation of diseased arteries) alternatives to remove and debulk atheromatous plaque are being explored. Among these, excimer laser angioplasty (ELA) has been developed as a modality that offers an alternative to percutaneous transluminal coronary angioplasty. The laser energy is delivered via fiberoptics to the target lesion and the plaque material is rendered into a gaseous state (vaporized) by breaking the molecular bonds. With proper training and patient selection, ELA has complication rates that are no higher than conventional balloon angioplasty. Preliminary data indicate that the laser is most successful in treating lesions that are not well treated with conventional balloon angioplasty. These include saphenous vein graft lesions, aorto-ostial lesions, long lesions (> 10 mm in length), moderately calcified stenoses, total occlusions, and percutaneous transluminal coronary angioplasty dilitation failures. The purpose of this article is to review the history and physics of ELA and to provide the nursing professional with an understanding of how the procedure is performed. Nursing considerations when caring for the patient undergoing this procedure are provided. PMID- 8420854 TI - Radiofrequency catheter ablation for supraventricular tachycardia. AB - Radiofrequency catheter-mediated ablation is a recently developed technique of achieving cure of certain rhythm disorders, notably supraventricular tachycardia. In less than a decade, it has evolved from a theoretic concept to first line therapy for many patients, including those in the pediatric age group. The most common types of arrhythmias amenable to catheter ablation include supraventricular tachycardia due either to an accessory pathway or A-V nodal reentry and atrial fibrillation with a rapid ventricular response refractory to medical therapy. Before the development of the technique, the only therapeutic options included either lifelong antiarrhythmic drugs or open heart surgery. The initial experiences with catheter ablative techniques used direct current shocks of up to 400 J, delivered to an intracardiac catheter from a standard defibrillator. This energy source has largely been replaced by radiofrequency current because of the substantially lower morbidity and greater efficacy. The treatment of patients with these tachyarrhythmias is evolving from a palliative approach to a curative one. With the current worldwide experience, complications are relatively rare and do not appear to differ from those associated with routine electrophysiologic testing or cardiac catheterization. The success rate is high, usually exceeding a 90% efficacy, but is dependent on the skill and experience of the operator. The technique, arrhythmias amenable to treatment, and the nursing implications before, during, and after the procedure are discussed in this article. PMID- 8420855 TI - Effects of child visitation in adult critical care units: a pilot study. AB - OBJECTIVE: To examine the behavioral and emotional responses of the child and of the nonhospitalized adult family member (NHAFM) to facilitated child visitation in the critical care setting. DESIGN: Quasi-experimental, posttreatment design. SETTING: An adult surgical intensive care unit at a large Midwestern teaching hospital. PARTICIPANTS: Twenty families participated in the study, 10 families in a restricted and 10 families in a facilitated visitation group. Each family unit had a child, an NHAFM, and a critically ill family member. In the control group the NHAFMs visited the patient in the customary routine, but children were restricted from visiting. After a 2-week waiting period a facilitated child visitation intervention was implemented for the experimental group. OUTCOME MEASURES: The child completed measures on anxiety as measured by the Manifest Anxiety Scale and behavioral and emotional changes as measured by the Perceived Change Scale. The NHAFM completed measures on anxiety as measured by the State Trait Anxiety Inventory and mood as measured by the Mood Adjective Check List. Family functioning, as measured by the Feetham Family Functioning Survey, and life event changes, as measured by the Life Event Scale, were examined as extraneous variables. INTERVENTION: The Child Visitation Intervention encompassed systematic facilitation and supervision of children visiting a critically ill adult family member in a surgical intensive care unit and provision of emotional support before, during, and after visitation (the intervention protocol may be obtained from the investigators on request). RESULTS: Children in the facilitated visitation group had a greater reduction (t = 4.0, df = 18, p = 0.0004) in negative behavioral and emotional changes as measured by the Child-Perceived Change Scale when compared with children in the restricted visitation group. CONCLUSION: Facilitated child visitation may help children deal with the critical illness of an adult family member and deserves further study. PMID- 8420856 TI - Child and family reactions during and after pediatric ICU hospitalization: a pilot study. AB - OBJECTIVE: To describe child behaviors and level of family functioning after discharge from the pediatric ICU (PICU) and to begin to explore the relationship of family reactions and the child's severity of illness to child and family outcomes after discharge. DESIGN: Exploratory, repeated measures: Time 1 (T1) at 24 hours after admission; Time 2 (T2) at 2 to 4 weeks after hospital discharge. SUBJECTS: Nine mothers and fathers of children who were 5 years of age or younger, hospitalized in a PICU for at least 1 day, expected to survive. MAIN STUDY MEASURES: Parental Concern Scale and Parental Stressor Scale: PICU at T1; Posthospitalization Behavior Questionnaire at T2; Feetham Family Functioning Survey and FACES III, at both T1 and T2; and Pediatric Risk of Mortality to measure illness severity. RESULTS: Mothers' cohesion scores decreased significantly from T1 to T2. Scores from the Pediatric Risk of Mortality were not related to family measures. Mothers' family cohesion and satisfaction with family after discharge were negatively related to time the child was intubated. Few family measures were related to the Parental Concern Scale and Parental Stressor Scale: PICU subscales. CONCLUSIONS: PICU admission of a child is a stressful event for parents, independent of the child's illness severity. Mothers' perceptions of family may be negatively affected. PMID- 8420857 TI - Effects of iced and room temperature injectate on cardiac output measurements in critically ill patients with low and high cardiac outputs. AB - OBJECTIVE: To determine the effect of injectate temperature (iced or room temperature) on cardiac output values in critically ill adults with low and high cardiac outputs. DESIGN: Quasi-experimental. SETTINGS: Two multidisciplinary intensive care units in two large, metropolitan, private, nonprofit hospitals in Texas. SUBJECTS: A convenience sample of 21 critically ill men and women who averaged 61 years of age (range 31 to 82 years) and whose most recent cardiac output measured with room temperature injectate was low (< or = 3.5 L/min) or high (> or = 8.0 L/min). INTERVENTION: Iced injectate and room temperature injectate (randomly ordered) were used to measure cardiac output in each subject. OUTCOME MEASURES: Cardiac output value with iced injectate versus cardiac output value with room temperature injectate. RESULTS: We found significant differences between cardiac output measurements with room temperature and those with iced injectate in eleven critically ill patients with low cardiac outputs (< or = 3.5 L/min) and in ten critically ill patients with high cardiac outputs (> or = 8.0 L/min). In the low cardiac output group, cardiac outputs using room temperature injectate averaged 0.37 L/min (range 0.1 to 1.10 L/min) higher than cardiac outputs using iced injectate (p = 0.001). In the high cardiac output group, measurements with room temperature injectate averaged 1.17 L/min L/min (range 0.3 to 3.0 L/min) higher than cardiac outputs with iced injectate (p = 0.005). Percent differences between room temperature and iced injectate values averaged 13% (range 3% to 27%) in patients with low cardiac outputs and 11% (range 3% to 29%) in patients with high cardiac outputs. Seven (77%) of the patients in the low cardiac output group and four (40%) of the patients in the high cardiac group had a 10% or greater difference--which many clinicians describe as a clinically significant difference--between room temperature and iced injectate cardiac output values. CONCLUSION: Although research is clearly needed to substantiate these findings, we suggest that nurses use iced injectate in patients with low and high cardiac outputs (< or = 3.5 L/min or > or = 8.0 L/min) to ensure accurate measurement of cardiac output. PMID- 8420858 TI - Symptoms, thoughts, and environmental factors in suspected acute myocardial infarction. AB - OBJECTIVE: To increase our understanding of patients' reactions and behavior at onset of symptoms of myocardial infarction. PROCEDURE: During a 5-month period a questionnaire focusing on symptoms, thoughts, and environmental factors at onset of symptoms was administered to all patients admitted to the coronary care unit at Sahlgrenska Hospital in Goteborg, Sweden, because of suspected acute myocardial infarction. RESULTS: A myocardial infarction developed in 48% of the 226 patients answering the questionnaire. In 81% of the patients, chest pain was the main symptom bringing them to the hospital. Forty-three percent characterized their symptoms as an oppression or uncomfortable feeling. Eighty-five percent suspected that the pain emanated from the heart, and yet 51% hesitated to go to the hospital, mainly because they expected the pain to disappear. Efforts to relieve pain were made by 63%, (mostly with nitroglycerin), which was taken more often by patients who did not subsequently develop a myocardial infarction than by those who did. Only 50% of the patients used an ambulance for transportation to hospital. There was a significant relation between subjective assessment of severity of symptoms and 1-year mortality (p < 0.05) and rehospitalization rate (p < 0.01), respectively. CONCLUSION: The majority of patients seem to interpret the symptoms of a myocardial infarction correctly and also have a correct perception of the severity of symptoms. For only a few is the natural next step to immediately call for an ambulance to get to the hospital. PMID- 8420859 TI - Transfer of a patient with a ventricular assist device to a non-critical care area. AB - Ventricular assist device (VAD) support has traditionally been associated with critically ill patients. Indeed, a VAD is inserted as the last hope for patients with cardiogenic shock who are unresponsive to conventional therapy. However, many patients bridged to potential cardiac transplantation are no longer critically ill after hemodynamic stabilization is achieved with VAD support. The focus of this article is to provide guidelines established for the transfer and provision of quality nursing care for patients with a VAD on a general cardiothoracic nursing floor. PMID- 8420860 TI - Restrained patients: an important issue for critical care nursing. AB - OBJECTIVE: To determine practice and attitudes of acute-critical care nurses toward the use of physical restraints. DESIGN: Survey with a self-administered anonymous questionnaire. SUBJECTS: A self-selected sample of 235 acute-critical care nurses from 17 states who were enrolled in critical care review classes. MEASUREMENTS: Data were gathered by use of a questionnaire developed by the researchers. The questionnaire was designed to elicit information regarding demographic and professional characteristics, nursing practice, and attitudes toward the use of restraints. Nurses responded to the practice items on a three point Likert Scale as to whether they "always," "sometimes," or "never" carried out the procedure. To obtain information on attitudes, nurses were asked to respond on a three-point Likert Scale as to whether they "agreed," were "undecided," or "disagreed" with the statement. RESULTS: Responses indicated 78% of the sample "always" try alternative nursing measures before restraining the patient. However, when units were believed to be understaffed there was more reliance on restraints. Thirty-eight percent of the sample would "always" rather sedate patients than physically restrain them. The overall score on the attitude component of the questionnaire reflected positive or acceptable attitudes toward the use of restraints. Most (62%) "agreed" that a patient suffers a loss of dignity when placed in restraints. Significant relationships did exist between select demographic and professional characteristics and practice and attitudes regarding the use of restraints. The longer the respondents had worked in critical care the more appropriate (positive) their attitudes toward the use of restraints (r = -.211, p < .01). CONCLUSIONS: In general, nursing practice and attitudes regarding the use of physical restraints indicated that respondents were using restraints in accordance with accepted practice. There is a need for additional research in this area, especially regarding the use of alternative measures to physical restraints. PMID- 8420861 TI - Recurrent unilateral lung hyperinflation as a manifestation of "auto-PEEP". AB - Recurrent right lung hyperinflation developed in a 68-year-old man with chronic airflow obstruction. His past history was significant for tuberculosis in the left lung requiring a left upper lobe resection and residual bronchiectasis in the remaining lung. Recognition of the unilateral lung hyperinflation as being a manifestation of unintentional positive end-expiratory pressure (auto-PEEP) resulted in appropriate measures to reverse the process and prevent further complications such as barotrauma. Patients with chronic airflow obstruction and an inflammatory or fibrotic disease process primarily involving one lung are at risk for the development of this atypical presentation of auto-PEEP. PMID- 8420862 TI - Spotlight article: does routine nursing care complicate the physiologic status of the premature neonate with respiratory distress syndrome? (Peters KL. J Perinat Neonat Nurs 1992;6[2]:67-84.). PMID- 8420863 TI - Over the top. PMID- 8420864 TI - Care of critically ill patients in the ED. PMID- 8420865 TI - The use of a multi-leaf collimator for conformal radiotherapy of carcinomas of the prostate and nasopharynx. AB - We investigate the use of a multi-leaf collimator for conformal radiation therapy of carcinomas of the prostate and of the nasopharynx. Following verification of dose calculation algorithms for multi-leaf collimated fields using film dosimetry, we compute dose distributions for multi-field conformal treatment using fields shaped with either the multi-leaf collimator or conventional cerrobend blocks. We compare the two sets of treatment plans using graphical isodose displays, tissue specific dose volume histograms, tumor control probabilities, and normal tissue complication probabilities. We also incorporate setup errors into the calculated dose distributions to assess the effect of treatment uncertainties on the various criteria. Based on these comparisons, we conclude that for multi-field conformal radiotherapy for these two disease sites, the use of multi-leaf collimation is equivalent to that of conventional cerrobend blocks. PMID- 8420866 TI - Initiation of multi-leaf collimator conformal radiation therapy. AB - Clinical studies have been initiated in conformal radiotherapy using a computer controlled multi-leaf collimator. Quantitative dosimetry and treatment planning studies comparing field shaping by lead alloy blocks and the multi-leaf collimator demonstrate the clinical acceptability of the multi-leaf collimator. Sixteen patients with tumors in multiple sites have received some part of their treatments with both blocking systems. Studies of dosimetry and field shaping show that the multi-leaf collimator produces clinically acceptable blocking for most field shapes and disease sites. The 80-20% penumbra was characterized for a wide range of shaped beams. For straight edges perpendicular to the leaf travel, the penumbra of measured dose distributions from the multi-leaf collimator is equal to conventional divergent blocking. When the multi-leaf collimator leaves approach a contour at an angle, the penumbra increases. At forty-five degrees, the maximum angle of approach, the penumbra is approximately 4 mm wider than that for divergent blocks. Three-dimensional treatment planning demonstrates that equivalent dose distributions can be obtained from the two field shaping systems. The multi-leaf collimator can be used effectively and efficiently to treat a variety of disease sites. Its optimal utility may be in treating complex fields- five or more shaped coplanar or non-coplanar beams. It is well suited for conformal therapy applications. PMID- 8420867 TI - Characterization of a multi-leaf collimator system. AB - Commissioning measurements for a multi-leaf collimator installed on a dual energy accelerator with 6 and 15 MV photons are described. Detailed dosimetric characterization of the multi-leaf collimator is a requirement for modeling the collimator with treatment planning software. Measurements include a determination of the penumbra width, leaf transmission, between-leaf leakage, and localization of the leaf ends and sides. Standard radiographic film was used for the penumbra measurements, and separate experiments using radiochromic film and thermoluminescent dosimeters were performed to verify that distortions of the dose distribution at an edge due to changing energy sensitivity of silver bromide film are negligible. Films were analyzed with a scanning laser densitometer with a 210 micron spot. Little change in the penumbra edge distribution was noted for different positions of a leaf in the field. Experiments localizing the physical end of the leaves showed less than 1 mm deviation from the 50% decrement line. This small difference is attributed to the shaped end on the leaves. One side of a single leaf corresponded to the 50% decrement line, but the opposite face was aligned with a lower value. This difference is due to the tongue and groove used to decrease between-leaf leakage. For both energies, approximately 2% of photons incident on the multi-leaf collimator are transmitted and an additional 0.5% leakage occurs between the leaves. Alignment of the leaves to form a straight edge results in a penumbra profile which compares favorably with the standard technique of using alloy blocks. When the edge is stepped, the isodose lines follow the leaf pattern and the boundary is poorly defined compared to divergent blocks. PMID- 8420868 TI - CT characteristics of patients with brain metastases treated in RTOG study 79-16. AB - The search continues for a favorable subgroup of patients with brain metastases in whom testing of new modalities might show a benefit in overall survival. Complete pre- and post-treatment CT evaluation of the brain was performed in 779 of the 859 patients entered into RTOG protocol 7916, a phase III study of the role of misonidazole combined with radiation therapy in the treatment of brain metastases. Pretreatment scan findings of mass effect, midline shift, massive edema, central necrosis, location of sentinel lesion, and number of lesions were correlated with length of survival for all patients as well as for each treatment group. The only characteristics that showed a statistically significant difference in survival in the overall group were the presence of < or = 3 lesions and the presence of a midline shift. The actual benefit in overall survival, however, was found to be only 3 weeks. The volume of the largest lesion prior to treatment did not correlate well with survival, nor did location of lesions. The time to response, number of responders and absolute decrease in number of lesions were similar for the four treatment arms. Patients who responded to cranial treatment had a significantly prolonged survival over those who did not respond. No CT characteristic evaluated in this study showed value as a clinically relevant prognosticator for patients with brain metastases for the overall group. Patients who fulfilled three of the four favorable clinical characteristics previously described by Diener-West (age < or = 60, KPS > or = 70, primary lesion absent or controlled and brain as sole site of metastasis), were analyzed separately. Those with < or = three lesions had a statistically significantly prolonged survival over those with four or more lesions. PMID- 8420869 TI - Optic glioma in children: surveillance, resection, or irradiation? AB - Eighty-seven consecutive children with newly diagnosed optic glioma were managed at University of Toronto hospitals 1958-1990. Overall the 10-year survival, relapse-free survival and freedom from second relapse rates were 84%, 68% and 85%. Twenty-seven patients relapsed or progressed, of whom 40% were free of a second relapse 10 years after the first relapse. Fourteen patients had a second relapse. Thirteen are dead. None survived 5 years after second relapse. Patients with anteriorly located tumors (N = 35), which involved the optic nerve, or chiasm and optic nerves, fared better than those with posteriorly located tumors (N = 52) with spread beyond the chiasm, 10-year survival 95% versus 76%, (p = .02), 10-year relapse-free survival 80% versus 59% (p = .02), respectively. For posterior tumors primary irradiation was more effective than primary subtotal resection for prevention of subsequent relapse, 10-year relapse-free survival 75% versus 41% (p = .02), but salvage therapy was, in part, successful and multivariate analysis of prognostic factors influencing survival for posterior tumors indicated that neither primary resection nor primary irradiation were significant factors. For first relapse, primary irradiation and the presence of neurofibromatosis were the significant favorable factors. Since 1977 and for posterior optic glioma subtotal resection or surveillance were used in 21/29 (72%) patients compared with 4/23 (17%) previously. Ten-year survival rates before and after 1977 were 78% and 67% and 10-year relapse-free survival 64% and 56%, respectively. PMID- 8420870 TI - Cranial nerve length predicts the risk of delayed facial and trigeminal neuropathies after acoustic tumor stereotactic radiosurgery. AB - PURPOSE: To test the hypothesis that length of cranial nerve irradiated is a major factor predicting the risk of cranial nerve injury following radiosurgery and to identify any other significant related treatment factors. METHODS AND MATERIALS: Ninety-two patients (93 acoustic tumors) were treated with a 201 source Cobalt-60 gamma unit from 1987 to 1990 and prospectively followed. The range of minimum tumor dose was 12-20 Gy and maximum dose 24-50 Gy. Univariate and multivariate analyses were used to evaluate any correlations between tumor measurements and treatment factors, with the development of trigeminal and facial neuropathies following radiosurgery. RESULTS: The risks of trigeminal and facial neuropathy following radiosurgery were associated with the pon-petrous distance and mid porous transverse tumor diameters respectively (anatomically related to the irradiated length of cranial nerves V and VII respectively) in both univariate (p = .002 for V and p = .026 for VII) and multivariate (p = .004 for V and p = .055 for VII) analyses. Tumor volume, other tumor measurements, maximum dose, minimum tumor dose, and tumor dose inhomogeneity were not significantly related to either trigeminal or facial neuropathy in univariate and multivariate analyses. CONCLUSION: Within a minimum tumor dose range of 12-20 Gy, the incidence of delayed trigeminal or facial neuropathy depended more on the estimated length of nerve irradiated than the tumor dose or tumor volume. In the future, the risk of delayed facial or trigeminal cranial neuropathy may be reduced significantly by performing radiosurgery when the tumor still has both a small mid-porous transverse diameter and a small pons-petrous distance. PMID- 8420871 TI - Contemporary management of adult and pediatric brain stem gliomas. AB - The aim of this study was to analyze the survival of a series of patients diagnosed as having brain stem tumors by computerized tomography scan, magnetic resonance imaging and/or biopsy and treated with megavoltage irradiation. Fifty three patients presenting to the Peter MacCallum Cancer Institute with a diagnosis of brain stem tumor from January 1980 to July 1989 were reviewed. There were 32 pediatric (age < or = 16) and 21 adult patients. The median age at presentation was 12 years (range 2-73 years). Eighteen patients had biopsy proof of glioma, the rest were diagnosed on the basis of CT and/or MRI appearance. Eighty-seven percent of patients received 44-55 Gy in 1.67-2.25 Gy fractions to the brain stem. Seventy-seven percent of patients showed improvement following treatment, 6% were stable, 11% progressed, and 6% were not evaluable. The estimated median survival from presentation for all patients was 34 months with 49% surviving at 3 years. The estimated median time to progression was 19 months with 34% not progressed at 3 years. There was no significant survival difference between patients with biopsy-proven glioma and those patients where the diagnosis was made radiologically. Survival was not significantly affected by age-group or sex. Patients with symptoms for more than 9 months prior to presentation had significantly longer survival than those with shorter duration of symptoms (p = 0.002). This paper presents the survival of patients with brain stem tumors diagnosed and treated by contemporary radiological and radiotherapeutic techniques. PMID- 8420872 TI - Effects of fractionated radiation therapy on human brain tumor multicellular spheroids. AB - We investigated the cytotoxic effects of fractionated radiation therapy on multicellular spheroids of human malignant glioma cell lines U-87 MG, U-251 MG, and U-373 MG. Graded doses of x-rays were administered in 1, 3, 8, 15, and 30 fractions over 15 days. The isoeffect dose for a 1 log cell kill ranged from 4 4.5 Gy for a single fraction to 7-8 Gy for an 8-fraction protocol; no additional dose-sparing was achieved with more fractions. Therefore, the effects of individual doses (1.56 Gy) of the 8-fraction protocol were studied in U-251 MG spheroids. A cell survival assay showed that the first dose of radiation killed 30-50% of the cells; subsequent doses usually killed fewer cells. The cell kill after all 8 doses was about 1.0 log. No consistent relationship between the intracellular glutathione level and fraction number was observed. The 24-hour labeling index of the spheroids did not decrease until after the second fraction. Thus, the higher cell kill of the first dose does not seem to be related to cell cycle synchrony. Multinuclear and mononuclear giant cells were limited almost entirely to the periphery of the spheroids and increased with the number of radiation fractions. We conclude that multicellular spheroids can be used to study the biological effects of fractionated radiation therapy on human brain tumor cells. Although this model cannot be used to evaluate the effect of radiation on normal tissue, it may be useful in developing more effective radiation therapy protocols for human brain tumors. PMID- 8420873 TI - The TCD50 and regrowth delay assay in human tumor xenografts: differences and implications. AB - The response to irradiation of five human xenograft cell lines--a malignant paraganglioma, a neurogenic sarcoma, a malignant histiocytoma, a primary lymphoma of the brain, and a squamous cell carcinoma--were tested in nude mice. All mice underwent 5 Gy whole body irradiation prior to xenotransplantation to minimize the residual immune response. The subcutaneous tumors were irradiated at a tumor volume of 120mm3 under acutely hypoxic conditions with single doses between 8 Gy and 80 Gy depending on the expected radiation sensitivity of the tumor line. Endpoints of the study were the tumor control dose 50% (TCD50) and the regrowth delay endpoints growth delay, specific growth delay, and the tumor bed effect corrected specific growth delay. Specific growth delay and corrected specific growth delay at 76% of the TCD50 was used in order to compare the data to previously published data from spheroids. The lowest TCD50 was found in the lymphoma with 24.9 Gy, whereas the TCD50 of the soft tissue sarcomas and the squamous cell carcinoma ranged from 57.8 Gy to 65.6 Gy. The isoeffective dose levels for the induction of 30 days growth delay, a specific growth delay of 3, and a corrected specific growth delay of 3 ranged from 15.5 Gy (ECL1) to 37.1 Gy (FADU), from 7.2 Gy (ENE2) to 45.6 Gy (EPG1) and from 9.2 Gy (ENE2) to 37.6 Gy (EPG1), respectively. The corrected specific growth delay at 76% of the TCD50 was correlated with the number of tumor rescue units per 100 cells in spheroids, which was available for three tumor lines, and with the tumor doubling time in xenografts (n = 5). The TCD50 values corresponded better to the clinical experience than the regrowth delay data. There was no correlation between TCD50 and any of the regrowth delay endpoints. This missing correlation was most likely a result of large differences in the number of tumor rescue units in human xenografts of the same size. PMID- 8420874 TI - A comparison of tumor and normal tissue microvascular hematocrits and red cell fluxes in a rat window chamber model. AB - This laboratory has previously used a window chamber model to measure red blood cell velocity in mammary tumors and normal granulation tissues of the F-344 rat. Because red cell flux and hematocrit more accurately reflect the oxygen carrying potential of blood, we used this model to measure these parameters. Red blood cells were labelled with fluorescein isothiocyanate, and 0.2 ml. packed cells were injected intravenously into rats bearing an 8 to 10 day old R-3230 mammary carcinoma. beta-phycoerythrin (0.15 mg.) was also injected and served as a plasma dye to outline the blood vessels. A sample of peripheral blood was then taken and analyzed by flow cytometry to determine the labeled fraction of red blood cells. Flowing tumor and normal tissue vessels were recorded onto a VCR, and these video images were used to determine vascular length and diameter, RBC flux and velocity, and hematocrit. Median vessel diameter and loge (red blood cell flux) were significantly greater in tumors than in normal tissues (p = 0.007 and p < 0.025, respectively). After controlling for these variables, the median tumor hematocrit of 19% was not significantly greater than the median normal tissue hematocrit of 15%. This technique provides a nontoxic and reproducible method that is now being used to assist in the in vivo definition of tumor oxygenation. PMID- 8420875 TI - Radiotherapy of the R1H-tumor: dose-rate effect on tumor response in brachytherapy with 106-ruthenium eye applicators. AB - PURPOSE: The influence of the dose-rate on tumor response in radiotherapy with 106-Ruthenium eye plaques has been investigated in an experimental tumor system. MATERIALS AND METHODS: The identical total dose was applied within three different overall treatment times (42 hr, 192 hr and 312 hr), corresponding to dose-rates of 6.0 Gy/hr, 1.3 Gy/hr and 0.8 Gy/hr. RESULTS: The therapeutic outcome of brachytherapy varied significantly between the three groups of animals treated with different dose-rates. At a dose-rate of 1.3 Gy/hr all tumors were locally controlled, but no local control was observed when a dose-rate of 6.0 Gy/hr was delivered. 0.8 Gy/hr was less effective than 1.3 Gy/hr but more effective than 6.0 Gy/hr. CONCLUSION: These results were unexpected but they might be explained by an incomplete reoxygenation if the overall treatment time is too short (42 hr, dose-rate 6.0 Gy/hr) and by proliferation of tumor cells under treatment if the overall treatment time is too long (312 hr, 0.8 Gy/hr). PMID- 8420876 TI - Radiobiological studies of PC-3 and DU-145 human prostate cancer cells: x-ray sensitivity in vitro and hypoxic fractions of xenografted tumors in vivo. AB - PURPOSE: To determine the quantitative responses to x-irradiation of exponentially growing human prostatic cancer cell lines PC-3 and DU-145 in vitro, and to determine the hypoxic percentages of these two cell lines when grown in vivo as xenografted solid tumors in nude mice. METHODS AND MATERIALS: Radiation survival in vitro was quantitated using both single-hit, multitarget and linear quadratic formalisms. Hypoxic fractions in vivo were determined from tumors of average sizes of about 750 mm3 using clonogenic excision assay. RESULTS: In vitro, the average single-hit, multitarget survival values for 7 replicate experiments for the DU-145 line were n = 1.92 (1.39-2.65), Dq(Gy) = 1.25, and Do(Gy) = 1.91 (1.88-1.94) (all values in parentheses indicate 95% confidence limits). For the PC-3 line (10 replicate experiments), these values were n = 2.84 (2.11-2.79), Dq(Gy) = 1.02, and Do(Gy) = 1.06 (0.87-1.25). For the linear quadratic formalism, values of alpha(Gy-1 x 10(1) and beta(Gy-2 x 10(2) for the DU-145 and PC-3 lines were, respectively, 1.55 (0.42) and 5.21 (1.09); and 4.87 (1.11) and 5.50 (1.88). The mean percentage survival of the DU-145 and PC-3 lines at a dose of 2 Gy were, respectively, 59.8 (53.3-67.0) and 32.0 (25.8-38.2). In vivo, the hypoxic fractions for the DU-145 and PC-3 tumors were, respectively, 7.20 (4.30-11.5), and 52.3 (42.8-63.9). RESULTS: The data from the in vitro experiments show that the DU-145 cell line is significantly more radioresistant than the PC-3 cell line. In vivo, the DU-145 tumors exhibit a significantly lower hypoxic percentage than do PC-3 neoplasms. CONCLUSIONS: Results indicate that significantly variability exists within human prostate tumors in regard to both intrinsic radiosensitivity in vitro and levels of hypoxia in vivo. Because these data appear to be the first published information on the intrinsic radiosensitivity and intratumor hypoxia characteristics of human prostate cancer, additional studies are needed to define the distributional aspects of these clinically relevant endpoints. PMID- 8420877 TI - Sensitivity of hyperthermia trial outcomes to temperature and time: implications for thermal goals of treatment. AB - PURPOSE: In previous work we have found that the cumulative minutes of treatment for which 90% of measured intratumoral temperatures (T90) exceeded 39.5 degrees C was highly associated with complete response of superficial tumors. Similarly, the cumulative time for which 50% of intratumoral temperatures (T50) exceeded 41.5 degrees C was highly associated with the presence of > 80% necrosis in soft tissue sarcomas resected after radiotherapy and hyperthermia. In the present work we have calculated the time for isoeffective treatments with T90 = 43 degrees C and T50 = 43 degrees C, respectively, using published thermal isoeffective dose formulae. The purpose of these calculations was to determine the sensitivity of treatment outcome to variations in thermal isoeffective dose. METHODS AND MATERIALS: The basis for the calculations were the thermal parameters and treatment outcomes in three patient populations: 44 patients with moderate or high grade soft tissue sarcoma treated preoperatively with hyperthermia and radiation; 105 patients with superficial tumors treated with hyperthermia and radiation, and 59 patients with deep tumors treated with hyperthermia and radiation. RESULTS: The thermal dose values calculated are strongly associated with outcome in multivariate logistic regression analysis. Simple dose-response equations result from the analysis, and we use these equations to assess the sensitivity of outcome upon variations in thermal dose. This information, in turn, allows us to estimate the number of patients required in Phase II and III trials of hyperthermia and radiation therapy. CONCLUSIONS: For regimens of 5 to 10 hyperthermia treatments, improvements in median T90 (superficial tumors) and T50 (deep tumors) parameters by 1.2-1.5 degrees C could result in response rates high enough (compared to radiotherapy alone) to justify Phase III trials. A similar improvement in response rates would require an increase in overall duration of treatment by a factor of 3 to 5. This would be difficult to achieve while also avoiding thermal tolerance induction. Achieving these temperature goals may be possible with improvements in hyperthermia technology. Alternatively, there may be ways to increase the sensitivity of cells to temperatures that can be achieved currently, such as pH reduction or chemosensitization. PMID- 8420878 TI - The role of the urethrogram during simulation for localized prostate cancer. AB - Urethrograms on 89 consecutive patients with localized prostate cancer were evaluated retrospectively, and the inferior border of the treatment field based on this study was compared with the inferior border that would have been defined by using the lower border of the ischial tuberosities. An analysis of the relationship between the margin used and the dose at the inferior border of the prostate supported our policy of requiring a 2 cm margin for optimal coverage of the prostate. Inclusion of at least 1 cm of proximal penile urethra was essential to ensure this 2 cm margin. Based on this assumption, twenty-five percent of patients would have had an inadequate margin if the lower border of the ischial tuberosities had been used instead of the urethrogram to define the inferior border of the treatment field. Assuming that a margin of more than 3 cm inferiorly is excessive, 11% of patients would have had excessive urethral irradiation if the bottom of the ischial tuberosities had been used to define the inferior border. Combining these two extremes, more than one in three patients would have had an inappropriate inferior margin if the bottom of the ischial tuberosities had been used to define the inferior border of the treatment field. There was no apparent increase in morbidity as a result of the urethrograms or an increase in treatment related toxicity in association with using the treatment fields defined by urethrography. Computed tomography was complimentary in defining the apex of the prostate. These data support the routine use of the urethrograms during simulation for localized prostate cancer. The use of the lower border of the ischial tuberosities to define the inferior border of the treatment field is associated with an unacceptable risk of either underdosing the apical portion of the prostate or overdosing the urethra. PMID- 8420879 TI - Treatment of arteriovenous malformations with stereotactic radiosurgery employing both magnetic resonance angiography and standard angiography as a database. AB - Twenty-one arteriovenous malformations were prospectively evaluated using magnetic resonance angiography and compared with stereotactic angiography. The goals were to establish the feasibility of magnetic resonance angiography, compare it to stereotactic angiography, employ magnetic resonance angiography in follow-up, and semiquantify flow. A correlative evaluation between flow and response to stereotactic radiosurgery was carried out. Phase contrast angiograms were obtained at flow velocities of 400, 200, 100, 60, and 20 cm/sec. The fractionated velocities provided images that selectively demonstrated the arterial and venous components of the arteriovenous malformations. Qualitative assessment of the velocity within the arteriovenous malformations and the presence of fistulae were also determined by multiple velocity images. In addition, 3-dimensional time-of-flight magnetic resonance angiograms were obtained to define the exact size and shape of the nidus. This technique also permitted evaluation of the nidus and feeding arteries for the presence of low flow aneurysms. Correlation between the two imaging modalities was carried out by subjective and semiquantitative estimation of flow velocity and estimation of nidus size. The following velocity parameters were employed: fast, intermediate, slow, and none (arteriovenous malformation obliterated). In 19 of 21 (90.5%) arteriovenous malformations, magnetic resonance angiography was equal or superior to stereotactic angiography for flow quantification and visualization of the nidus. Only 2 of 21 arteriovenous malformations were better demonstrated by stereotactic angiography than by magnetic resonance angiography (failure rate of 9.5%). The nidus size in one case was clearly underestimated by stereotactic angiography and would have resulted in a geographic miss without magnetic resonance angiography. Seven post-radiosurgery arteriovenous malformations were evaluated for follow-up with both magnetic resonance angiography and stereotactic angiography. In 6 of 7 arteriovenous malformations, magnetic resonance angiography response matched stereotactic angiography response. Correlation of flow with outcome was carried out for 14 arteriovenous malformations using magnetic resonance angiography only. Interestingly, all nine arteriovenous malformations with intermediate or slow flow demonstrated partial or complete obliteration; whereas only 3 of 5 fast flow arteriovenous malformations achieved a response with a median follow-up of 10 months. This early analysis suggests that slower flowing arteriovenous malformations may obliterate faster after stereotactic radiosurgery and flow parameters could be employed to predict response. In conclusion, magnetic resonance angiography permits semiquantitative flow velocity assessment and may therefore be superior to stereotactic angiography. An additional advantage of magnetic resonance angiography is the generation of serial transverse images which can replace the conventional CT scan employed for stereotactic radiosurgery treatment planning.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8420880 TI - Gamma knife surgery of cerebral arteriovenous malformations: serial MR imaging studies after radiosurgery. AB - PURPOSE: To investigate the temporal sequence of post radiosurgery magnetic resonance imaging changes in cerebral arteriovenous malformations. METHODS AND MATERIALS: Eighteen patients were regularly followed up after gamma knife surgery. The follow-up intervals ranged from one day to 44 months. High signal lesion in or around arteriovenous malformations on T2-weighted magnetic resonance images corresponding to the treatment volume and developing after radiosurgery were defined as the adverse reaction of the irradiation. This high signal and the regression of arteriovenous malformations nidus after radiosurgery were evaluated. RESULTS: Adverse reaction of irradiation was observed in nine cases. Seven of them were symptomatic. The reactions presented as focal high signal in three cases and focal high signal with extension along the neural tracts in six cases. The reactions were seen either immediately after treatment (one case), between 3 and 14 months (seven cases), and 40 months after treatment (one case). The regression of the adverse reaction was observed to start 5 +/- 3 months after its appearance. Regression of the arteriovenous malformations' nidus was found in 16 cases. In two cases the AVMs became invisible on magnetic resonance images but the angiogram still demonstrated abnormal shunts. In another one case with angiogram showing total obliteration, the nidus was erroneously interpreted as incomplete obliteration on magnetic resonance images. CONCLUSION: It is concluded that magnetic resonance imaging is a sensitive in vivo method for detecting cerebral radiation injury. Magnetic resonance imaging offers a method for evaluating the regression of arteriovenous malformations' nidus, but the diagnosis of complete obliteration of the nidus after radiosurgery still relies on the angiogram. PMID- 8420881 TI - Continuous low dose rate brachytherapy with high activity iodine-125 seeds in the management of meningiomas. AB - The purpose of this study was to evaluate if meningiomas can be effectively treated with brachytherapy using permanent implantation of high activity I-125 seeds. Thirteen patients with intracranial meningiomas were treated by means of permanent stereotactic implantation of one or more high-activity I-125 seeds. The physical characteristics of I-125 enabled us to deliver a minimum tumor dose ranging from 100 Gy to 500 Gy at a low dose rate of 5 cGy to 25 cGy per hr. Indications for this procedure included recurrence after initial surgery or as primary modality of treatment in patients who were not candidates for surgery. All 13 patients are alive at a median follow-up of 25 months. Nine of 13 patients achieved complete resolution of the tumor and in the remaining four, more than 50% reduction in tumor volume was noted at the last follow-up. No late complications were observed. We conclude from this initial data that localized high dose irradiation delivered at a low dose rate using I-125 permanent implantation is an effective, safe, and simple method in the treatment of both recurrent and primary intracranial meningiomas. PMID- 8420882 TI - Poor outcome in radiation-induced constrictive pericarditis. AB - PURPOSE: To compare the outcome of patients with radiation-induced constrictive pericarditis versus patients with constriction due to another etiology. METHODS AND MATERIALS: Twenty patients with constrictive pericarditis seen during 1975 1986 at a single medical center. Six had radiation-induced constrictive pericarditis (Group A). The etiology was idiopathic in ten subjects and secondary to carcinomatous encasement, chronic renal failure, purulent infection and tuberculosis in one patient each (Group B, N = 14). Mean age was 53.4 +/- 15.5 years. RESULTS: Extensive pericardiectomy was performed in 3/6 Group A and 13/14 Group B patients. All Group A patients died, 4 weeks--11 years post-diagnosis (median = 10 months). Two Group A patients died suddenly, one died post operatively of respiratory failure, another of pneumonia and two of recurrent carcinoma. Thirteen Group B patients are alive (median follow-up = 72 months). The only death in this group was due to metastatic cancer. CONCLUSION: The poor outcome with radiation-induced constriction is probably multi-factorial. Poor surgical outcome is to be expected in patients with evidence of recurrent tumor, high-dose irradiation, pulmonary fibrosis or associated radiation-induced myocardial, valvular or coronary damage. PMID- 8420883 TI - Portal film enhancement: technique and clinical utility. AB - We report on the results a 3-year project which had as its goal the development of methods to enhance radiation portal films to improve their readability. We had previously reported on a portal film enhancement technique, contrast limited adaptive histogram equalization, which could enhance low contrast detail, but degraded sharply contrasted edges. A new method, unsharp masking followed by contrast limited adaptive histogram equalization, now appears to overcome this problem. A clinical trial to test whether enhanced portal films could be read more accurately than standard ones was undertaken. The trial involved 12 readers from two institutions doing 276 readings. In this trial the enhanced films were judged to be of higher quality than the non-enhanced films (p < .001) and were read more accurately (p = .026). The usefulness and difficulties of routinely performing portal film enhancement in a busy radiation therapy department are discussed. PMID- 8420884 TI - Prostatic thermoluminescent dosimeter analysis in a patient treated with 18 MV X rays through a prosthetic hip. AB - External beam radiation therapy with high energy photon beams through hip prostheses has been shown to cause dose inhomogeneities for target volumes in the pelvis. In this work, measurements of dose using thermoluminescent dosimetry were compared with dose calculations from a computerized treatment planning system in a patient with prostatic carcinoma and a cobalt-chromium-molybdenum hip prosthesis. A 39% decrement in dose at isocenter was demonstrated for an 18 MV photon beam passing through the prosthesis. A discrepancy of only 3.1% was shown between measured and calculated dose when the tissue-maximum ratio (TMR) method of heterogeneity correction was used. However, it is recognized that several sources of error are possible when heterogeneity corrections are performed for high density prostheses and these are discussed below. The results of this work stress the importance of accurate data for use with the "ratio of TMR's" algorithm in order that accurate treatment planning can be performed. PMID- 8420885 TI - A survey of radiation oncologists regarding their radiation physics instruction. AB - The American Association of Physicists in Medicine, Committee on Training of Radiologists conducted a survey of radiation oncologists requesting information regarding their radiation oncology physics training. General questions were asked of the oncologist regarding their radiation oncology practice such as number of oncologists, number of new patients treated, and the size and type of facility in which the practice is located. The oncologist also responded to questions regarding their educational background. The survey requested the radiation oncologists to answer questions regarding the adequacy and importance of their training in specific areas of radiation physics. The responders indicated that the importance of most physics topics in their clinical practice corresponded to the level of their understanding. The survey indicated that for most radiation oncologists their physics instruction was an important and interesting part of their residency program. PMID- 8420886 TI - Acute toxicity during external-beam radiotherapy for localized prostate cancer: comparison of different techniques. AB - PURPOSE: The chronic and acute toxicities associated with conventional radiotherapy of localized prostate cancer are well documented. However, the degree and incidence of toxicities with conformal techniques are not known. Studying side effects associated with modern radiotherapeutic techniques is more important now since there has been a general trend to use computerized tomography based techniques in recent years; beam's eye view-based conformal techniques are also becoming more commonplace. It is possible that the local disease control can be improved with the delivery of higher doses than currently used. Conformation of the treatment volume to the target volume may facilitate such dose-escalation. However, prior to such dose-escalation, it is important to know the toxicities associated with such techniques with conventional doses. METHODS AND MATERIALS: We have compared week-by-week acute toxicities associated with conventional (Group A, 16 patients), computerized tomography-based, manual (Group B, 57 patients) and beam's eye view-based (Group C, 43 patients) techniques during 7 weeks of radiotherapy. Group B and C patients were treated contemporaneously (1988-1990). RESULTS: Acute side effects gradually increased from week 1 through weeks 4-5 and generally declined or plateaued after that. The incidence of acute toxicities was significantly less with the beam's eye view/based technique than with the other two methods. For instance, the percentages of Grade 2 acute genitourinary toxicities for Groups A, B, and C were as follows: Week 1-0, 0, 0; Week 2-6, 0, 0; Week 3-6, 9, 2; Week 4-12, 14, 9; Week 5-35, 14, 9; Week 6-31, 16, 7; Week 7-33, 8, 8, respectively. The p values associated with differences in acute genitourinary toxicities for Weeks 1-7 using chi-square test were 0.072, 0.627, 0.389, 0.538, 0.123, 0.06, and 0.012; the p values for acute gastrointestinal toxicities were 0.512, 0.09, 0.031, 0.031, 0.003, < 0.0001, and 0.004, respectively. Pairwise comparison (Wilcoxon rank-sum test) showed statistically significant lower acute toxicity in Group C than Group B (e.g., p values, Weeks 1-7 for gastrointestinal toxicity: 0.633, 0.056, 0.010, 0.014, < 0.0001, < 0.0001, and < 0.0001, respectively) in the latter part of the treatment course. No correlation was found between the extent of toxicity and the patient age or the overall treatment time. Also, no correlation was found between the degree of toxicity and the radiation dose and fraction size, within the narrow ranges used (65-70 Gy and 180-200 cGy, respectively). A trend suggesting increased severity of toxicity with increase in the volume of treatment was seen. CONCLUSION: The findings in this retrospective study need to be confirmed by other prospective studies. PMID- 8420887 TI - Optimization of radiation therapy and the development of multi-leaf collimation. PMID- 8420888 TI - Conformal radiation in prostate cancer: reduced morbidity with hope of increased local control. PMID- 8420889 TI - Response to Esche et al. PMID- 8420890 TI - Potentiating effect of diabetes in radiation retinopathy. PMID- 8420891 TI - The radiobiology of radiosurgery: rationale for different treatment regimes for AVMs and malignancies. AB - Based on basic radiobiological principles, we suggest that the radiosurgery technique of delivering a radiation dose in a single fraction, whilst appropriate for benign brain lesions such as arteriovenous malformations (AVM), is not optimal for treating malignant tumors. Radiosurgery was originally developed to treat benign lesions in the brain, such as AVMs, and has been successfully used for this purpose for over four decades. Recently, the technique has been adopted for treating small primary malignant brain tumors or single metastases. We argue, and derive radio-biological data to support the view that, treating malignant tumors with a single fraction will result in a suboptimal therapeutic ratio between tumor control and late effects, even for small tumors; and that improved therapeutic ratios would be expected if the treatment were fractionated into a small number of fractions. On the other hand, no therapeutic gain is to be expected from fractionating treatment of AVMs. A new generation of noninvasive relocatable stereotactic head frames makes feasible the use of fractionated stereotactic external-beam radiotherapy, and may allow significant benefits over single, radiosurgical, treatments for malignant brain tumors. As stereotactic fractionation/protraction regimes become more widespread, a uniform approach for determining equivalent fractionation schemes becomes important for intercomparing clinical results, and such calculations can be reliably carried out using the linear-quadratic formalism. PMID- 8420892 TI - Segmental ischemic necrosis of the small intestine in two postparturient mares. AB - Two mares developed segmental ischemic necrosis of the small intestine after parturition. In one mare, the mesentery of the small intestine apparently tore during parturition, after which necrosis developed in a 52-cm segment of the distal portion of jejunum. In the other mare, a 52-cm segment of the proximal portion of ileum tore during parturition. Three weeks later, a 40-cm segment of the distal portion of jejunum became incarcerated in the mesenteric rent and twisted 360 degrees on its mesentery. Both mares recovered after resection of affected intestine and are functioning as broodmares. PMID- 8420893 TI - Postoperative evaluation of renal function after surgical correction of a ureterovaginal fistula in a cat. AB - Unilateral hydronephrosis and urinary incontinence, secondary to a ureterovaginal fistula, were corrected in a cat after surgical excision of the fistula and implantation of the affected ureter into the bladder. Salvage of the kidney was attempted because underlying chronic bilateral renal disease was suspected. Renal scintigraphy was used to monitor improvement in the function of the previously obstructed kidney after surgery. PMID- 8420894 TI - Renal failure attributable to atrophic glomerulopathy in four related rottweilers. AB - Atrophic glomerulopathy resulting in chronic renal failure was diagnosed in 4 related Rottweilers, each < 1 year old. All 4 dogs had severe azotemia and massive protein-losing nephropathy. Histologically, the glomerular lesion was characterized by mild dilatation of Bowman's space, with glomerular tufts absent or markedly atrophied. The lesion is distinct from the congenital glomerular changes described in Samoyeds or Doberman Pinschers. PMID- 8420895 TI - Jejunal intussusception in adult horses: 11 cases (1981-1991). AB - Medical records of 11 adult horses with jejunal intussusception examined at 5 veterinary teaching hospitals between 1981 and 1991 were reviewed. Nine of 11 horses had signs of acute abdominal discomfort for < 24 hours, whereas 2 horses had a history of chronic signs. Five of 11 horses had an intraluminal or intramural mass associated with the jejunal intussusception. Two horses died or were euthanatized prior to surgery. Partial jejunal resection and jejunojejunal anastomosis were performed in 9 horses. One horse died during surgery and 2 were euthanatized prior to hospital discharge because of postoperative complications. Four of the 6 horses that were discharged from the hospital survived from 16 months to 6 years and returned to their previous level of performance. One horse died 3 months after surgery from unknown causes, and 1 horse was lost to long term follow-up evaluation. PMID- 8420896 TI - Congenital Neospora infection in calves born from cows that had previously aborted Neospora-infected fetuses: four cases (1990-1992). AB - Four dairy cows that had been successfully rebred following fetal Neospora infection and abortion were identified from 2 drylot dairies. All 4 cows had uncomplicated pregnancies with the birth of 5 full-term calves. The calves all had high precolostral serum IgG antibodies. The precolostral antibodies to Neospora sp as determined by indirect fluorescent antibody test ranged from 5,120 to 20,480, compared with maternal serum and colostral antibody titers from 320 to 1,280. Two calves had mild neurologic limb deficits. Three calves had mild nonsuppurative encephalomyelitis and Neospora organisms were found in the CNS of 3 calves. Findings indicate that repeat transplacental Neospora infections occur in cows. Additionally, calves born from cows with a history of Neospora fetal infection and abortion may have congenital Neospora infections and/or neurologic dysfunctions at birth. The Neospora indirect fluorescent antibody test appears to be a useful antemortem test for detection of calves exposed in utero to Neospora organisms. PMID- 8420898 TI - Steel-jaw leghold trap. PMID- 8420897 TI - High-rise syndrome in dogs: 81 cases (1985-1991). AB - We evaluated 81 dogs with high-rise syndrome. Dogs fell from 1 to 6 stories, and of 52 dogs for which the fall was witnessed, 39 had (75%) jumped. Dogs sustained a triad of injuries to the face, thorax, and extremities, similar to injuries seen in cats with high-rise syndrome, but with differences in degree and distribution. Height fallen and landing surface affected initial status and type and severity of injury. Cause of fall influenced distribution of extremity injury. Dogs falling < 3 stories had a high prevalence of extremity fractures. Higher falls resulted in more spinal injuries. We recommend initial treatment for shock and thoracic trauma followed by orthopedic and neurologic evaluation. Visceral trauma should be considered if response to emergency treatment is poor. All but 1 of the dogs survived. PMID- 8420899 TI - Tympanic bulla osteotomy for treatment of middle-ear disease in cats: 19 cases (1984-1991). AB - Twenty-two tympanic bulla osteotomies were performed in 19 cats for the treatment of bacterial otitis media (n = 11), inflammatory polyps (n = 7), or middle-ear neoplasia (n = 4). Inflammatory polyps mostly affected young adult cats (mean age, 1.5 years), whereas otitis media affected middle-aged cats (mean age, 5.5 years) and neoplasia affected older cats (mean age, 10.25 years). Although not specific for the underlying disease process, fluid density within the tympanic bulla or thickening of the bulla was observed radiographically in 100% of cats with middle-ear disease. In cats for which results were available, bacterial culturing revealed pathogens in 6 cats diagnosed with otitis media and in 7 cats with benign or malignant tumors within the bulla. The ventral approach to the bulla was the most common surgical procedure (18 osteotomies), followed by the lateral approach combined with ablation of the vertical and horizontal parts of the external acoustic meatus (4 osteotomies). Immediate postoperative complications were common but short-term, and included Horner's syndrome (11 cats), facial nerve paralysis (5 cats), and otitis interna (2 cats). Bulla osteotomy resulted in resolution of clinical signs without recurrence in cats with middle-ear polyps. Drainage of the infected tympanic cavity aided in resolution of the clinical signs of bacterial otitis media, but the long-term clinical course was characterized by persistent neurologic deficits (5 cats) and recurrence of disease (2 cats). Neoplastic infiltration of the tympanic bulla was associated with a poor prognosis, and although surgery helped to establish a diagnosis, it did not alter the clinical course of the disease.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420900 TI - Hepatic fatty cirrhosis in ruminants from western Texas. PMID- 8420901 TI - What is your diagnosis? Hemangiosarcoma of the falciform ligament. PMID- 8420902 TI - Neospora threatens livestock, dogs. PMID- 8420903 TI - Pieces of scrapie puzzle falling into place. PMID- 8420904 TI - From a satisfying hobby to a fulfilling, full-time job. PMID- 8420905 TI - Residue prevention strategies in the United States. PMID- 8420906 TI - Indemnifying veterinary medical association board members and officers. PMID- 8420907 TI - Anesthetic techniques for neutering 6- to 14-week-old kittens. AB - Forty-eight male and 48 female 6- to 14-week-old kittens were neutered by use of 4 anesthetic protocols. Preanesthetic disposition, depth of sedation, loss of resistance to handling, induction quality, induction time, sternal and stand times, and recovery quality were evaluated. Analgesia and muscle relaxation without supplemental inhalational anesthetics were evaluated in male kittens, and the time until extubation was recorded in female kittens. Intramuscular administration of tiletamine/zolazepam (TZ), midazolam/ketamine, atropine/midazolam/ketamine/butorphanol (AMKB), and atropine/midazolam/ketamine/oxymorphone (AMKO) produced rapid sedation and smooth induction into anesthesia. In male kittens, there were no significant differences in sedation, relaxation, induction time, or quality. Tiletamine/zolazepam administration induced the best analgesia, and midazolam/ketamine administration induced the least analgesia for castration. The recovery time in male kittens was longest with TZ and shortest with the opioid groups (AMKB, AMKO). In females, TZ produced significantly faster induction times, but the degree of sedation and relaxation after administration of injectable agents was not significantly different among the groups. More females given TZ could be intubated without supplemental inhalational agents than females in other groups. Extubation time was rapid in all groups, but the times until sternal and standing were significantly longer, and recovery quality was significantly poorer in females given TZ. In kittens given opioids, reversal of the opioid did not shorten recovery time or improve recovery quality.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420908 TI - Prevalence and morphologic features of apical deltas in the canine teeth of dogs. AB - Prevalence and morphologic features of the apical delta in the canine teeth of dogs were determined to assist clinicians in the endodontic treatment of these teeth. Eighty-two canine teeth from mixed-breed dogs were cleared and dyed to outline root canal ramifications. Apical deltas were classified according to location in the dental arch, number of ramifications, and vertical extent of these ramifications from the apical terminus. A classification system was devised to aid in identification and treatment of the delta. Recommendations were made regarding indications for conventional and/or surgical endodontic therapy depending on the type of delta encountered. Apical deltas were detected in 69.5% of the canine teeth studied. Surgical endodontic intervention should be seriously considered when the delta is found to extend a distance > 1 mm from the apical terminus. PMID- 8420909 TI - Comparison of adverse effects of phenylbutazone, flunixin meglumine, and ketoprofen in horses. AB - The relative toxicity of phenylbutazone, flunixin meglumine, and ketoprofen was studied in healthy adult horses. Sixteen horses were randomly assigned to receive 10 ml of physiologic saline solution, or ketoprofen (2.2 mg/kg of body weight), flunixin meglumine (1.1 mg/kg), or phenylbutazone (4.4 mg/kg) IV, every 8 hours, for 12 days. Results of CBC, serum biochemical analyses, and fecal occult blood tests were monitored. On day 13, all horses were euthanatized and complete necropsy examinations were performed. Mean CBC values remained within normal limits for all groups. Phenylbutazone-treated horses had a significant (P < 0.05) decrease in serum total protein and albumin concentrations. Mean values of all other serum biochemical assays were not different from those of the saline treated group. Results of all fecal occult blood tests were negative. At necropsy, the glandular portion of the stomach was the area of the gastrointestinal tract most severely affected by phenylbutazone, flunixin meglumine, and ketoprofen. In the phenylbutazone-treated group, but not in the other groups, edema of the small intestine and erosions and ulcers of the large colon were observed. None of the horses treated with saline solution had lesions in the glandular portion of the stomach or in the intestine. Four horses (1/5 and 3/3 in the flunixin- and phenylbutazone-treated groups, respectively) developed renal crest necrosis. Horses in the saline- and ketoprofen-treated groups did not develop renal lesions.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420910 TI - Prevalence and severity of lameness in lactating dairy cows in a sample of Minnesota and Wisconsin herds. AB - An epidemiologic study investigating the prevalence of lameness in lactating dairy cows was performed in 17 dairy herds in Minnesota and Wisconsin. The mean herd size was 50 cows. Cows from 14 herds were housed in stanchions or tie stalls, and cows from 3 herds in free stalls or dry lot. During visits to each farm in the summer and subsequent spring, 2 investigators simultaneously but independently evaluated the ambulation of lactating cows by use of a standardized scoring system. The lameness scoring system was reliable at the 2 visits, with 92.7 and 91.3% agreement between the 2 observers and kappa coefficients of 0.60. The prevalence of lameness detected by the investigators ("clinical" lameness) was 13.7% (117/853) in summer and 16.7% (134/801) in spring in lactating dairy cows. These prevalences were 2.5 times higher than those estimated by the herd managers. Parity was significantly (P < or = 0.03) associated with lameness, with higher prevalence of clinical lameness found in cows of higher parity. PMID- 8420911 TI - Aldrin intoxication and clearance of associated dieldrin residues in a group of feedlot cattle. AB - A sudden onset of bizarre neurologic dysfunction was found in 8 of 90 mixed-breed feeder calves. Seven other calves were dead, and 3 more died during the next week. A diagnosis of organochlorine toxicosis was made when rumen and abomasal contents from 1 of the calves revealed 22.4 and 20.6 micrograms of aldrin/g of ingesta, respectively. Complete feeds retrieved from self-feeders contained 54 and 528 micrograms of aldrin/g of feed. The initial concentration range in fat from 40 live calves was 6.01 to 42.44 micrograms of dieldrin/g of fat. Additional fat samples were analyzed to verify residue compliance until the entire herd was clear of residue 18 months after removal of the contaminated ration. The range of apparent half-lives for dieldrin in body fat of heifers and steers was 69 to 231 and 53 to 116 days, respectively. These findings demonstrate the considerable variability in apparent half-life of dieldrin in field cases. In cases of dieldrin-contaminated livestock, veterinarians and regulatory personnel must accurately determine the necessary slaughter withholding times so that informed economic decisions are made in the best interest of the producer while enhancing the probability of a safe food supply. Excretion rates of dieldrin from field contaminated cattle may not be consistent with results obtained under experimental conditions. PMID- 8420912 TI - Motor neuron degeneration in a horse. AB - A 9-year-old Quarter Horse mare was examined because of progressive weight loss, weakness, muscle atrophy and tremors, and behavioral change. Selenium and glutathione peroxidase assays, blood lead analysis, erythrocyte transketolase analysis, pseudorabies and Borrelia burgdorferi serology, electromyography, and CSF analysis were performed. Motor neuron degeneration was diagnosed by microscopic examination of neural tissues. The cause of the disease was not substantiated, but several possibilities were excluded via diagnostic testing. Diagnosis of motor neuron degeneration in horses may be made from an accurate history, thorough neurologic examination, and ancillary testing. In particular, antemortem diagnosis may be based on finding scattered angular atrophy of predominantly type-1 or of type-1 and -2 skeletal muscle fibers in frozen sections of muscle biopsy specimens. PMID- 8420913 TI - Treatment of smoke inhalation in five horses. AB - Five horses were admitted for treatment of smoke-inhalation injuries sustained in a barn fire. Three of the horses were mildly affected, with high respiratory rates (24 to 36 breaths/min) and normal to low arterial oxygen tensions (77.0 to 94.1 mm of Hg), and responded well to administration of diuretics, bronchodilators, corticosteroids, and antibiotics. The 2 remaining horses were severely affected. Both were in respiratory distress, with markedly low arterial oxygen tensions (50.4 and 57.1 mm of Hg) and cyanosis. These 2 horses required fluid resuscitation in addition to the treatments given to the less severely affected horses. Tracheostomy was performed to facilitate removal of large, obstructive, pseudomembranous tracheobronchial casts. Oxygen was administered by nasal or tracheal insufflation or by use of a high-frequency jet ventilator. The most severely affected horse developed hemorrhagic colitis and was euthanatized. The 4 surviving horses recovered in 2 to 5 months and resumed working without reduction in performance capability. PMID- 8420914 TI - Osteosarcoma in a cow. AB - A mass involving the maxilla of a cow was determined to be an osteosarcoma. Microscopically, the mass was composed of plump pyriform cells aligned along thin trabeculae of osteoid. Osteosarcoma rarely develops in large animal species, but when it does, it is usually found in the skull and must be differentiated from more common diseases, such as actinomycosis and tooth abscesses. PMID- 8420915 TI - Complications of middiaphyseal radial ostectomy performed for treatment of premature closure of the distal radial physis in two dogs. AB - Middiaphyseal radial ostectomy was performed in 2 dogs that had premature closure of their distal radial physes. Complications included disuse osteoporosis, carpal hyperextension of the affected and contralateral limbs, degenerative joint disease involving the cubital and carpal joints, synostosis of the radius to the ulna, premature closure of the distal portion of the ulna, and failure to achieve normal limb length. Both dogs were lame after exercise. PMID- 8420916 TI - Binding and degradation of lectins by components of rumen liquor. AB - The binding of 15 different plant lectins to feed particles and microbes in rumen liquor, and their degradation were studied in vitro. The rate of degradation assessed from the label released when radioactive iodine-labelled lectins were incubated with rumen liquor conflicted with the rates calculated from measurements of the survival of the antigenic structure (immuno-rocket electrophoresis) or the biological function (haemagglutination) of the lectins. Thus solubilization of the radioactive label indicated that Concanavalin A (Con A), but not the soyabean agglutinin, SBA, or kidney bean phytohaemagglutinin, PHA E3L, was stable to rumen proteolysis. In contrast, both SBA and PHA-E3L were shown by immuno-rocket electrophoresis or haemagglutination tests to be highly resistant to breakdown, while the degradation of Con A proceeded at a constant slow rate under the same conditions. This was in accord with the previously established general stability of lectins in the gut of single-stomach animals. Of the 15 lectins, SBA, favin (Vicia faba lectin) and Con A were bound by hay and the particle fraction of rumen liquor. This was, in part, specific and reversible in the presence of appropriate sugars. Most pure bacterial strains preferentially bound lectins with specificity for glucose/mannose (favin and Con A), while rumen fungi reacted with SBA. The level of binding was low with other lectins. However, inter-strain differences of lectin-binding were found in Selenomonas ruminantium and Ruminococcus flavefaciens. Clearly, as some lectins were not fully degraded in the rumen, they could be expected to depress the utilization of the diet not only in single-stomach animals but, possibly, also in ruminants. PMID- 8420917 TI - Distribution of pathogen inhibition in the Lactobacillus isolates of a commercial probiotic consortium. AB - Pure strains of Lactobacillus ssp. isolated from a commercial probiotic consortium were checked in a double layer solid medium for their inhibition activities against selected pathogenic bacteria including serotypes of Listeria monocytogenes, Escherichia coli and Salmonella. The antagonistic properties of the Lactobacillus strains may be related to the production of bacteriocin-like compounds. All the pathogens tested were inhibited by one or a few strains of Lactobacillus, the best inhibition was observed against L. monocytogenes but the inhibition was also satisfactory against E. coli, Salm. typhimurium and Salm. enteritidis. PMID- 8420918 TI - Biosynthesis of aflatoxin--the role of nutritional factors. PMID- 8420919 TI - Detection by in vitro amplification of the alpha-toxin (phospholipase C) gene from Clostridium perfringens. AB - A polymerase chain reaction (PCR) with thermostable DNA polymerase from Thermus aquaticus is described for the specific amplification of the phospholipase C (alpha-toxin) gene of Clostridium perfringens. A set of primers selected for their high specificity could detect Cl. perfringens in stools with a detection limit of approximately 5 x 10(2) bacteria, after bi-amplification. A modified PCR without thermal steps was performed to rapidly amplify, with a yield of 60%, the DNA template. With this PCR method Cl. perfringens alpha-toxin gene could be detected within 2 h. The PCR method detected alpha-toxin positive Cl. perfringens but did not react with phospholipase C-producing Bacillus cereus, Pseudomonas aeruginosa, Cl. sordellii and Cl. bifermentans. The amplified PCR products were screened through ethidium bromide agarose gel electrophoresis or, in only 1 h, with the PhastSystem (Pharmacia). This PCR satisfies the criteria of specificity, sensitivity and rapidity required for a useful tool in epidemiology and for the diagnosis of the pathogen Cl. perfringens as it may be used directly on stool samples. PMID- 8420920 TI - Purification, partial characterization and plasmid-linkage of pediocin SJ-1, a bacteriocin produced by Pediococcus acidilactici. AB - Pediococcus acidilactici SJ-1, isolated from a naturally-fermented meat product, produced an antibacterial agent active against selected strains of Lactobacillus spp., Clostridium perfringens and Listeria monocytogenes. The agent was bactericidal against sensitive indicators, and sensitive to proteolytic enzymes; it was identified as a bacteriocin, and was designated as pediocin SJ-1. It was stable over a wide pH range (3-9), and apparently most stable in the lower part of that range. At pH 3.6, pediocin SJ-1 was stable at heat-processing temperatures within the range 65-121 degrees C; its activity decreased significantly, however, when it was heated at pH 7.0. The activity of pediocin SJ 1 on sensitive indicator cells was lost in the presence of alpha-amylase, suggesting that it contains a glyco moiety, necessary for its antibacterial action. Native pediocin SJ-1 exists in the form of monomers and aggregates (with molecular weights in the range 80-150 kDa). Pediocin SJ-1 was purified 262-fold by direct application of cell-free supernatant fluids to a cation-exchange chromatography column, and was resolved by SDS-PAGE as a single peptide band with a MW of ca 4 kDa. The original pediocin SJ-1-producing strain (bac+) harbours three plasmids of 4.6, 23.5, and 45.7 MDa. Production of pediocin SJ-1, but not immunity to SJ-1, is associated with the 4.6 MDa plasmid. PMID- 8420921 TI - The effect of slurry storage and anaerobic digestion on survival of pathogenic bacteria. AB - The decline in viable numbers of Salmonella typhimurium, Yersinia enterocolitica and Listeria monocytogene in beef cattle slurry is temperature-dependent; they decline more rapidly at 17 degrees C than at 4 degrees C. Mesophilic anaerobic digestion caused an initial rapid decline in the viable numbers of Escherichia coli, Salm. typhimurium, Y. enterocolitica and L. monocytogenes. This was followed by a period in which the viable numbers were not reduced by 90%. The T90 values of E. coli, Salm. typhimurium and Y. enterocolitica ranged from 0.7 to 0.9 d during batch digestion and 1.1 to 2.5 d during semi-continuous digestion. Listeria monocytogenes had a significantly higher mean T90 value during semi continuous digestion (35.7 d) than batch digestion (12.3 d). Anaerobic digestion had little effect in reducing the viable numbers of Campylobacter jejuni. PMID- 8420922 TI - Adaptation of Pseudomonas aeruginosa to 2,2'-methylenebis (4-chlorophenol). AB - A culture of Pseudomonas aeruginosa, isolated from a cooling water system, was grown in the presence of sub-inhibitory concentrations of 2,2'-methylenebis(4 chlorophenol) (MBC). It adapted to increasing concentrations from an initial minimum inhibitory concentration of 36 micrograms ml-1 to the highest, 80 micrograms ml-1. Resistant cultures exhibited a higher survival rate when exposed to 320 micrograms ml-1 than did the original strain. Lipopolysaccharide and outer membrane protein profiles were determined by SDS PAGE. No changes were detected in lipopolysaccharide profiles. The quantity of OprP, the phosphate uptake protein in the outer membrane, decreased to a low level correlating with decreased phosphate (P(i)) uptake during growth. It is proposed that OprP is the place of entry for MBC and that the cell can adapt by decreasing the level of OprP in the outer membrane. PMID- 8420923 TI - The MARCKS family of cellular protein kinase C substrates. PMID- 8420924 TI - Promoter activity of human renin 5'-flanking DNA sequences is activated by the pituitary-specific transcription factor Pit-1. AB - Although the renal juxtaglomerular cell is the source of circulating renin, the renin gene is also expressed at a number of extrarenal sites including lactotrope cells of human and ovine pituitaries. In the present study, we demonstrate that GC cells, a pituitary lactotrope precursor cell line, efficiently express transiently transfected hybrid genes containing human renin 5'-flanking DNA sequences -148/+11. Gel mobility shift competition analyses show that a highly conserved sequence in human and rodent renin 5'-flanking DNAs (human coordinates: -80/-58) binds a nuclear factor from GC cells, most likely the pituitary-specific factor Pit-1. Deletional and mutational analyses demonstrate that this site is a major determinant of renin promoter activity in GC cells. Transfection of a Pit-1 expression construct into HeLa cells, where activity of the human renin promoter is low, stimulates expression of cotransfected renin-luciferase constructs. Moreover, activation of the human renin promoter by co-expression of Pit-1 is dependent on an intact Pit-1 site. Taken together, these data strongly suggest that Pit-1 activates pituitary renin gene expression. This finding raises the possibility that member(s) of the POU family of transcription factors, of which Pit-1 is an archetypal member, may direct renin expression in other tissues, including the kidney. PMID- 8420925 TI - Cloning and expression of a mammalian Na+/amino acid cotransporter with sequence similarity to Na+/glucose cotransporters. AB - We describe the full-length sequence and functional expression of a cDNA cloned from LLC-PK1 cells, which appears to encode a mammalian Na(+)-dependent neutral amino acid transporter with properties characteristic of system A. This sequence, designated SAAT1, is 76% identical and 89% similar in amino acid sequence to the Na(+)-dependent glucose transporter SGLT1 of the same species. A leucine zipper region was detected in both SAAT1 and SGLT1. The message for SAAT1 was a single 2.4-kilobase species in kidney, but mRNA species of 2.4 and 3.7 kilobases were observed in LLC-PK1 cells as well as in intestine. Transcripts were also found in spleen, liver, and muscle. Expression of SAAT1 in COS-7 cells resulted in increased levels of Na(+)-dependent uptake of 2-(methylamino)isobutyric acid, a specific substrate for the system A amino acid transporter. Uptake due to cDNA expression was inhibited by a range of amino acids that are transported by system A and exhibited a km of 0.8 +/- 0.2 mM. These results suggest that the system A amino acid transporter is closely related to the Na+/glucose transporter SGLT1. PMID- 8420926 TI - Disruption of potential sites for N-linked glycosylation does not impair hormone binding to the lutropin/choriogonadotropin receptor if Asn-173 is left intact. AB - The rat lutropin/choriogonadotropin receptor (rLHR) is a G protein-coupled receptor, the large extracellular domain of which binds human choriogonadotropin (hCG) with high affinity. Within the extracellular domain are six potential sites for N-linked glycosylation. Although several studies have attempted to determine if N-linked carbohydrates are necessary for hormone binding, the results have been in apparent disagreement. In this study we have used site-directed mutagenesis to singly and collectively alter the consensus sequences for N-linked glycosylation in the rLHR. In particular, we examined the binding activity in both intact cells as well as detergent-solubilized extracts so that the effects on trafficking to the plasma membrane could be determined. In addition, we independently examined the effects of substituting a particular Asn versus Thr or Ser residue within a given glycosylation consensus sequence. Our data suggest that substitution of Asn-173 with Gln results in both a decreased ability of the receptor to be expressed on the plasma membrane as well as a vastly decreased binding affinity of the receptor for hCG. However, if the consensus sequence for N-linked glycosylation at Asn-173 is altered by substitution of Thr-175 with Ala (instead of Asn-173 to Gln), the resulting receptor binds hCG with high affinity although it is still impaired in its ability to be expressed at the plasma membrane. Furthermore, if all consensus sequences for N-linked glycosylation are mutated collectively while maintaining Asn-173 (by substituting Thr-175 with Ala instead of Asn-173 to Gln), the resulting deglycosylated receptor, although not expressed on the plasma membrane, binds hCG with high affinity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8420927 TI - The biological relevance of the binding of calcium ions by inositol phosphates. AB - The binding of Ca2+ (chelation) by myo-inositol polyphosphates at pH 7.0 was studied using a Ca(2+)-sensitive electrode. Glucose 6-phosphate (used as a model for a monophosphate) bound Ca2+ with an affinity of 152 +/- 31 liters/mol and a molar ratio of 0.94 +/- 0.02. Inositol 3,4-bisphosphate, inositol 1,4,5 trisphosphate, inositol 1,3,4,5-tetrakisphosphate, and inositol hexakisphosphate showed affinities of 9.0 +/- 2.1 x 10(3), 6.3 +/- 1.5 x 10(3), 6.2 x 10(4), and 1.92 +/- 0.47 x 10(5) liters/mol, respectively, and molar ratios of 0.92 +/- 0.49, 0.95 +/- 0.10, 0.75, and 2.5 +/- 0.5. In general, the affinity increased with the number of phosphate substituents on the inositol ring, although the stereochemistry is also expected to be important. This suggests that for the physiologically relevant inositol phosphates (tris-, tetrakis-, pentakis-, and hexakis-) half-maximal Ca2+ binding will occur in the Ca2+ concentration range of approximately 5 x 10(-6) to 2 x 10(-4) M. This range lies between the basal intracellular and the fee extracellular Ca2+ levels (10(-7) and 10(-3) M), respectively, and may therefore be of physiological importance. Chelation provides a possible simple explanation for the inhibition by Ca2+ of inositol 1,4,5-trisphosphate binding to its receptor in rat cerebellum and other tissues. It may also have a role in limiting inositol phosphate-mediated increases in intracellular Ca2+. PMID- 8420928 TI - Conformational, thermodynamic, and stability properties of Manduca sexta apolipophorin III. AB - Apolipophorin III (apoLp-III) is a major protein in hemolymph of adult Manduca sexta. Although it normally exists in a lipid-free state, during sustained flight, apoLp-III functions as an apolipoprotein, reversibly associating with the surface of lipoprotein particles. In an effort to gain a fuller understanding of this dual existence, we have investigated its solution properties using spectroscopic methods. The UV absorption spectrum of apoLp-III is distinctive owing to the absence of tryptophan and the presence of a single tyrosine residue. Circular dichroism experiments revealed an enhancement of apoLp-III alpha-helical content when spectra were obtained in 50% trifluoroethanol versus aqueous buffer. The helical content in buffer was unaffected by protein concentration, suggesting that apoLp-III exists in solution as a monomeric species. At pH values > 10 and < 4, there was a marked loss of helical content. Increasing the temperature of apoLp-III solutions also caused a loss of secondary structure, with a temperature induced denaturation midpoint of 52 degrees C. Upon recooling of heat-denatured apoLp-III, approximately 95% of the secondary structure was restored. In guanidine HCl denaturation studies monitored by CD, a 50% transition midpoint of 0.355 M was determined, corresponding to a delta GDH2O of 1.29 kcal/mol. Fluorescence studies indicated that guanidine HCl induced an enhancement of tyrosine fluorescence emission at 300 nm when excited at 277 nm. In native apoLp III, we propose that tyrosine fluorescence is quenched to a large extent due to a hydrophobic stacking interaction of its side chain with that of a neighboring phenylalanine residue. delta GDH2O was determined from the fluorescence data to be 2.1 kcal/mol, with a transition midpoint occurring at 0.25 M guanidine HCl. The lower concentration of guanidine HCl required to induce half-maximal tyrosine fluorescence enhancement versus the transition midpoint detected by CD may be a reflection of the fact that this residue is located near the COOH-terminal end of the protein and as such may be more susceptible to denaturation. The results presented indicate that apoLp-III assumes a relatively labile conformation in solution that appears to be partially stabilized by side chain charge-charge interactions within predicted alpha-helical segments. PMID- 8420929 TI - Evidence that energization of the chloroplast ATP synthase favors ATP formation at the tight binding catalytic site and increases the affinity for ADP at another catalytic site. AB - Previous results have not established whether the attainment of a rapid photophosphorylation rate as ADP concentration is increased in the micromolar range (apparent Km = approximately 30 microM) results from the filling of a second or a third catalytic site. Measurements reported here show that the ATP synthase of chloroplast thylakoids, with 2-4 microM medium ADP present during steady-state photophosphorylation, has one catalytic site filled with tightly bound nucleotides, but other catalytic sites are largely empty. Thus, the rapid increase in the photophosphorylation rate with higher ADP concentrations results from the filling of a second catalytic site. Even with 30 microM added ADP in the dark, the binding of more than one ADP per synthase was not detectable. The sensitivity of the assay was such that the Kd for binding of ADP at a second catalytic site of the de-energized synthase is > 150 microM, considerably above the apparent Km for rapid photophosphorylation. This result can be explained by an increase in the affinity of a second catalytic site for ADP upon energization. Other experiments assessed the effect of ADP binding at a second catalytic site on the equilibrium between bound ATP and ADP and P(i) at the tight catalytic site. When the rate of photophosphorylation is limited by a low ADP concentration, about equal amounts of ATP and ADP are bound at one catalytic site on the synthase. In contrast, when the rate is limited by a low P(i) concentration with 100 microM ADP present, the equilibrium of bound reactants is shifted so that close to one ATP per synthase is present. This is as expected if the binding of ADP at a second catalytic site allows the protonmotive force to promote ATP formation from ADP and P(i) at a tight binding catalytic site. A scheme for the binding change mechanism incorporating these results is presented. PMID- 8420930 TI - Slow binding of ATP to noncatalytic nucleotide binding sites which accelerates catalysis is responsible for apparent negative cooperativity exhibited by the bovine mitochondrial F1-ATPase. AB - The bovine heart mitochondrial F1-ATPase depleted of nucleotides (nd-MF1) hydrolyzes 50 microM ATP in three kinetic phases at 30 degrees C. An initial "burst" rapidly transforms into an intermediate, slower rate, which slowly accelerates to the final, steady-state rate. The intermediate phase disappears progressively as the concentration of ATP in the assay medium is increased and is absent at 2 mM. Activation in the intermediate phase is lost when nd-MF1 is inactivated by 5'-p-fluorosulfonylbenzoyladenosine, which modifies three noncatalytic sites. Correlation of [3H]ATP binding to nd-MF1, after treatment either with 50 microM Mg[3H]ATP plus a regenerating system or 10 mM free [3H]ATP, with stimulation of the intermediate phase suggests that this phase is abolished when at least two noncatalytic sites are filled with ATP. Prior incubation of nd MF1 with MgPPi stimulates hydrolysis of 30 microM to 2 mM ATP and abolishes the intermediate phase. Following incubation with Mg[32P]PPi, 3.3 mol of [32P]PPi/mol of enzyme are bound, 1 and 0.5 mol of which are released by cold chases with MgATP and MgITP, respectively. Since the cold chases diminish activation only slightly, the stimulatory effect is not caused by PPi binding to catalytic sites. A Lineweaver-Burk plot of initial rates of the intermediate phase for hydrolysis of 30 microM to 2 mM ATP by nd-MF1 is biphasic, extrapolating to apparent Km values of 120 and 440 microM. The latter value is the same as the apparent Kd determined from dependence of the rate of activation of the intermediate phase on ATP concentration in the assay medium. After prior incubation of nd-MF1 with MgPPi or free ATP, Lineweaver-Burk plots are linear with the highest Km disappearing. Thus, this Km reflects rate acceleration when ATP binds to noncatalytic sites. From these results it is concluded that slow binding of ATP to noncatalytic sites during hydrolysis of low concentrations of substrate, which accelerates catalysis, is responsible for apparent negative cooperativity exhibited by MF1. PMID- 8420931 TI - Cell penetration of diphtheria toxin. Reduction of the interchain disulfide bridge is the rate-limiting step of translocation in the cytosol. AB - The pathway of cell penetration of diphtheria toxin (DT) was studied in Vero cells by following the kinetics of uptake, reduction, degradation, and sub cellular distribution of 125I-DT in the absence or presence of bafilomycin A1 (baf-A1), a powerful inhibitor of the endosomal H(+)-ATPase. After a lag phase of 4 min, DT, bound to Vero cells, reached an acidic intracellular compartment, where about one-third of it underwent a transition to a state competent for subsequent reduction and membrane translocation. After further 4 min, this DT fraction was reduced in a baf-A1-insensitive reaction and DT-A, the intracellularly active protomer of DT, was immediately released in the cytosol. The present data indicate that cell-mediated reduction of the toxin is the rate determining step of the DT cell intoxication process. Subcellular fractionation showed that DT underwent the low pH-driven conformational change in an early endosome, distinct from the subsequent endosomal compartment where reduction took place. DT-B remained endosome-bound and was proteolyzed at low pH as well as the portion of DT which was not reduced after the exposure to low pH in early endosomes. PMID- 8420932 TI - Genetic and immunological analyses of the cyanobacterium Synechocystis sp. PCC 6803 show that the protein encoded by the psbJ gene regulates the number of photosystem II centers in thylakoid membranes. AB - The psbJ gene is a member of the psbEFLJ gene cluster in the cyanobacterium Synechocystis sp. PCC 6803 as well as in the chloroplasts of green plants. The putative product of the psbJ gene is a 4-kDa protein with one membrane-spanning domain. We have raised rabbit antibodies against a T7 gene 10-psbJ fusion protein, overexpressed in Escherichia coli. These antibodies recognized a polypeptide of expected size in the thylakoid membrane from wild type Synechocystis cells. We have also created a targeted mutant of Synechocystis 6803 in which the fourth codon of the psbJ open reading frame was modified to a translational stop codon. Thylakoid membranes from these mutant cells lacked the protein recognized by the antibodies. In the mutant cells, the partial electron transfer reaction mediated by the photosystem I complex was unaffected, whereas the rate of the photosystem II (PSII)-mediated reaction was 46% of that in wild type cells. Herbicide binding assays indicated that the PSII to chlorophyll ratio in the mutant cells was 49% of that in wild type cells. These results indicate that while the PsbJ protein is not essential for the photochemical activity it controls the amount of functionally assembled PSII complex in the thylakoid membrane. PMID- 8420933 TI - Histamine induces a gene-specific synthesis regulation of secretogranin II but not of chromogranin A and B in chromaffin cells in a calcium-dependent manner. AB - The effect of histamine on steady-state mRNA levels of three soluble secretory proteins from chromaffin granules was investigated in cultured bovine adrenal medullary cells. Histamine stimulated secretogranin II mRNA 4-fold, with no or only slight effects on chromogranin A and B transcription. After an initial lag phase, secretogranin II mRNA increased markedly between 4 and 12 h followed by a plateau phase up to 48 h. The effect of histamine on secretogranin II gene expression was abolished by the H1 receptor antagonist promethazine but not by the H2 blocker cimetidine. The histamine-induced elevation of secretogranin II mRNA was partially reduced by the L-type calcium channel blocker nifedipine, indicating a contribution of extracellular calcium in the second messenger signaling pathway involved. At present, the H1 histaminergic receptor is the first membrane receptor found regulating secretogranin II biosynthesis in chromaffin cells. The selective up-regulation of secretogranin II mRNA but not of chromogranin A and B by histamine is another example illustrating that the synthesis regulation of secretory components co-stored in large vesicles of neuroendocrine cells is specific to the individual gene. Together with results obtained earlier, our data demonstrate a marked variation in the relative composition of peptides secreted from adrenal medulla into circulation following different physiologic conditions. PMID- 8420934 TI - Mutations of the molecular chaperone protein SecB which alter the interaction between SecB and maltose-binding protein. AB - SecB is a 16-kDa cytosolic chaperone protein that is required for efficient export of particular proteins in Escherichia coli. To identify regions of SecB that contribute to efficient protein export, we isolated secB point mutants that are defective for protein export in vivo. We obtained missense mutations at residues Leu75 (SecBL75Q), Cys76 (SecBC76Y), and Glu77 (SecBE77K) in the center of the secB gene. In vivo, mutant SecBL75Q and SecBE77K proteins are capable of binding to precursor maltose-binding protein (MBP) and preventing the formation of export-incompetent precursor MBP; however, export of MBP is still defective. In vitro, purified SecBL75Q and SecBE77K proteins bound to unfolded MBP and blocked its refolding. SecBL75Q and SecBE77K were more effective than wild-type SecB at blocking the refolding of unfolded MBP, suggesting that SecBL75Q and SecBE77K have a higher affinity for unfolded MBP. PMID- 8420935 TI - The number of amphipathic alpha-helical segments of apolipoproteins A-I, E, and A IV determines the size and functional properties of their reconstituted lipoprotein particles. AB - The objective of this work was to determine the role of the amphipathic alpha helical structural units of human apolipoproteins A-I, E, and A-IV in defining the sizes and reactivities with lecithin:cholesterol acyltransferase (LCAT) of their reconstituted lipoprotein particles. We prepared reconstituted high density lipoprotein (rHDL) particles with each of the three apolipoproteins in two weight ratios with lipid: 2.7/0.07/1 and 1.35/0.04/1, palmitoyloleoylphosphatidylcholine/cholesterol/apolipoprotein, by the sodium cholate dialysis procedure; and examined the rHDL product sizes and distributions by nondenaturing gradient gel electrophoresis. The rHDL particles were also incubated with low density lipoprotein (LDL), and with LDL plus LCAT, to observe any structural modifications due to phospholipid transfers to LDL and to cholesterol esterification by LCAT. In addition, we examined the average structural properties of the original rHDL by several fluorescence methods and circular dichroism spectroscopy, and determined their reaction kinetics with LCAT. The results indicate that the diameters of the largest rHDL particles, containing two apolipoproteins per particle, correlate with the maximum number of putative amphipathic alpha-helical segments in their sequences, and that smaller particles of this class may arise from the removal of one or more alpha-helical segments from contact with lipid. Furthermore, the larger particles may be converted into the smaller ones upon loss of phospholipid to LDL, and may form one or two well defined products when reacted with LCAT. In general, the subclasses of particles have distinct spectroscopic properties, consistent with a different apolipoprotein folding in particles containing different proportions of phospholipid to apolipoprotein. Furthermore, the different apolipoprotein structures lead to significant differences in reactivity with LCAT. PMID- 8420936 TI - Alpha- and beta-xylosides alter glycolipid synthesis in human melanoma and Chinese hamster ovary cells. AB - beta-D-Xylosides are often used to competitively inhibit proteoglycan synthesis by serving as primers for free glycosaminoglycan (GAG) chain assembly. Quite unexpectedly, we found that when human melanoma cells and Chinese hamster ovary cells are labeled with [3H] galactose in the presence of 4-methyl umbelliferyl beta-D-xyloside (Xyl beta 4MU), a large portion of the labeled acceptor does not consist of the expected GAG chains, but of the novel GM3 ganglioside-like structure: Sia-alpha 2,3-[3H]Gal beta 1, 4Xyl beta 4MU. Moreover, formation of this derivative is associated with an inhibition of glycosphingolipid synthesis by up to 78% without affecting synthesis of other [3H]Gal-labeled glycoconjugates. Inhibition occurs rapidly and equally for all glycolipid species and is partially abrogated by brefeldin A. Inhibition requires the addition of a single galactose residue to the xyloside within the lumen of the Golgi apparatus. This addition appears to be carried out by galactosyl transferase I that normally synthesizes the core region of GAG chains. Although alpha-xyloside does not inhibit proteoglycan synthesis, it is galactosylated, but not sialylated, and is nearly as effective as a beta-xyloside at inhibiting glycolipid biosynthesis. Similar results were obtained for human macrophage U937, and differentiated or undifferentiated PC12 cells. However, in neuroblastoma cell line MR23, no low molecular weight xyloside products were made and glycolipid synthesis was not inhibited. These results suggest that some of the previously documented effects of beta-xylosides might result, in part, from their inhibition of glycolipid synthesis. The mechanism of inhibition is not a direct competition for glycolipid synthesizing enzymes; rather, it is an unexplained result of formation of Gal beta 1,4Xyl-1 (alpha or beta)4MU. PMID- 8420937 TI - Identification of vicinal thiols of phosphoenolpyruvate carboxykinase (GTP). AB - Phosphoenolpyruvate carboxykinase (PEPCK) from the cytosol of rat liver has 13 cysteines, at least one of which (Cys288) is known to be very reactive and critical for catalytic activity (Lewis, C. T., Seyer, J. M., and Carlson, G. M. (1989) J. Biol. Chem. 264, 27-33). Previous results provided evidence for the existence of at least 1 pair of vicinal cysteines within or near the active site of PEPCK (Lewis, C. T., Haley, B. E., and Carlson, G. M. (1989) Biochemistry 28, 9248-9255). An intramolecular cystine disulfide is induced to form upon treatment of PEPCK with equimolar 5,5'-dithiobis(2-nitrobenzoate) (Nbs2) or upon irradiation of the enzyme in the presence of the photoaffinity probe 8-azidoGTP. In each case, modification is accompanied by a substantial loss in catalytic activity, and substrates protect against inactivation and modification. We now report the identification of these modified thiols by differential alkylation of cysteines and half-cystines with radioactive iodoacetate, followed by isolation and sequencing of the modified tryptic peptides. The results indicate that the disulfide formed by equimolar Nbs2 lies within a 15-residue region of the PEPCK sequence that includes Cys399, Cys407, and Cys413. In addition, Cys407 and/or Cys413 also appear to participate in formation of the disulfide induced by 8 azidoGTP. These thiols lie very near a consensus sequence that has been suggested to represent the binding site for the guanine ring of GTP. PMID- 8420938 TI - Horseradish peroxidase-catalyzed two-electron oxidations. Oxidation of iodide, thioanisoles, and phenols at distinct sites. AB - The atypical two-electron oxidation of thioanisole and its p-methyl, p-methoxy, and p-nitro analogues by horseradish peroxidase, contrary to earlier reports, stereoselectively produces the (S) sulfoxides in 60-70% enantiomeric excess. Horseradish peroxidase reconstituted with delta-meso-ethylheme has little peroxidase (guaiacol oxidizing) activity, as previously reported, but exhibits increased sulfoxidation activity. Difference spectroscopy shows that guaiacol binds to delta-meso-ethylheme-reconstituted horseradish peroxidase even though it is essentially not oxidized. In contrast, horseradish peroxidase reconstituted with delta-meso-methylheme is active in both reactions. Studies with H(2)18O2 show that the oxygen in the sulfoxide produced by delta-meso-ethylheme reconstituted horseradish peroxidase derives, as it does in the reaction catalyzed by the native enzyme, primarily from the peroxide. Preincubation of horseradish peroxidase with phenylhydrazine, which modifies the protein, suppresses peroxidase activity but does not inhibit thioanisole sulfoxidation. On the other hand, the oxidation of iodide is blocked by reconstitution of horseradish peroxidase with delta-meso-ethylheme or preincubation with phenylhydrazine. Noncompetitive kinetics are observed for the inhibition of guaiacol and iodide oxidation by thioanisole and of guaiacol oxidation by iodide. The kinetic data and the differential inhibitory effects of delta-meso-ethylheme reconstitution and phenylhydrazine preincubation indicate that thioanisole and iodide, both of which undergo net two-electron oxidations, are oxidized at sites distinct from each other and from that involved in the oxidation of guaiacol. Spectroscopic substrate binding studies provide support for distinct thioanisole, guaiacol, and iodide-binding sites. An active site model is proposed to rationalize the results. PMID- 8420939 TI - The Streptococcus sanguis platelet aggregation-associated protein. Identification and characterization of the minimal platelet-interactive domain. AB - Streptococcus sanguis expresses a cell wall-bound protein that induces the activation and aggregation of platelets. This platelet aggregation-associated protein (PAAP) contains a collagen-like, platelet-interactive domain within a 23 kDa protein fragment. To isolate the minimal platelet-interactive domain, p23 PAAP was digested with collagenase, and the digest chromatographed to isolate fractions with activity inhibitory to S. sanguis-induced platelet aggregation. The active fraction was then digested with cyanogen bromide, the product chromatographed, and a smaller inhibitory peptide isolated. Finally, this fraction was digested with endoproteinase Lys-C, and the digest fractionated. After each step, inhibitory activity resolved into single chromatographic peaks of 13 kDa (p13 PAAP), 2.7 kDa (p2.7 PAAP), and a minimal 7-mer peptide, respectively. These PAAP fragments showed similar ID50 (19-28 nM), suggesting that each contained a single copy of the platelet-interactive domain. The minimal 7-mer peptide was purified by immunoaffinity chromatography and reverse phase high pressure liquid chromatography. The primary structure was determined to be Pro-Gly-Glu-Gln-Gly-Pro-Lys. This sequence conforms to the predicted structural motif of the platelet-interactive domains of types I and III collagen. This 7-mer peptide is therefore the platelet-interactive domain of the PAAP from S. sanguis. Its structure explains the molecular basis for immunological cross-reactivity and functional similarity to the platelet-interactive domains of collagens. PMID- 8420940 TI - Repair of individual DNA strands in the hamster dihydrofolate reductase gene after treatment with ultraviolet light, alkylating agents, and cisplatin. AB - We have analyzed gene-specific and strand-specific DNA damage and repair in the dihydrofolate reductase gene in hamster cells. Cells were UV-irradiated or treated with two types of chemotherapeutics, alkylating agents or cisplatin. UV induced pyrimidine dimers were detected using a previously published technique in which the T4 endonuclease V enzyme is used to create nicks at the lesion sites. 6 4 photoproducts were detected in a similar assay using ABC excinuclease after prior reversal of the pyrimidine dimers with photolyase. Adducts formed by the alkylating agents nitrogen mustard and dimethyl sulfate were quantitated by generating strand breaks at basic sites after neutral depurination. Cisplatin induced intrastrand adducts were detected with ABC excinuclease, and cisplatin interstrand cross-links were detected using a denaturation-reannealing reaction before electrophoresis. In accord with previous reports by other investigators, we find distinct strand specificity of the repair of pyrimidine dimers after UV; the transcribed strand was much more efficiently repaired than the nontranscribed strand. In contrast, there was little or no strand bias in the repair of the 6-4 photoproducts. For alkylating agents, a slight bias toward repair in the transcribed strand was found after treatment with nitrogen mustard, but there appeared to be no bias in the repair after treatment with dimethyl sulfate. Cisplatin interstrand cross-links are repaired with equal efficiency from the two strands, but the more common cisplatin-induced lesion, the intrastrand adduct, is preferentially repaired from the transcribed strand. In conclusion, there is strand bias in the repair of pyrimidine dimers and cisplatin intrastrand adducts, but the strand specificity of repair may not be a general feature for all DNA lesions, as we found little or no strand bias in the repair of other lesions studied. PMID- 8420941 TI - Phycobilins of cryptophycean algae. Novel linkage of dihydrobiliverdin in a phycoerythrin 555 and a phycocyanin 645. AB - Cryptomonad strain IVF2 phycoerythrin 555 carries phycoerythrobilins attached through single thioether bonds at alpha-Cys-18, beta-Cys-82, and beta-Cys-158 and a doubly linked 15,16-dihydrobiliverdin (DBV) at beta-DiCys-50,61 (for sequence numbering, see Sidler, W., Nutt, H., Kumpf, B., Frank, G., Suter, F., Brenzel, A., Wehrmeyer, W., and Zuber, H. (1990) Biol. Chem. Hoppe-Seyler 371, 573-547). Analysis of the beta-DiCys-50,61-linked DBV by 1H homonuclear and 1H-13C heteronuclear NMR spectroscopy establishes that the thioether bond from Cys-50 is to the 3"-carbon of the DBV ring A and that from Cys-61 is to the 18'-carbon of ring D, i.e. the peptide-linked bilin is an 8,12-bis(2-carboxyethyl)-3-(2 (cysteinyl-S)-ethyl)-18-(1-(cysteinyl-S)-e thyl)- 2,7,13,17-tetramethylbiladiene ab-1,19(16H,21H)-dione. DBV is also present at beta-DiCys-50,61 in cryptomonad strain UW374 phycocyanin 645 (Wedemayer, G. J., Kidd, D. G., Wemmer, D. E., and Glazer, A. N. (1992) J. Biol. Chem. 267, 7315-7331). NMR spectroscopy shows that the thioether bonds to this DBV are also at 3" and 18'. Linkage of tetrapyrroles to polypeptides through the 3"-carbon has not hitherto been reported. PMID- 8420942 TI - Uptake of high density lipoprotein cholesterol ester by HepG2 cells involves apolipoprotein E localized on the cell surface. AB - High density lipoprotein (HDL) cholesterol ester (CE) is taken up by many cells without simultaneous uptake of HDL apoprotein. The studies described herein demonstrate that the selective uptake of cholesterol ester by HepG2 cells is reduced by antibody directed against the receptor-binding domain of apoE (monoclonal antibody (mAb) 1D7) but not by antibody directed against the NH2 terminal portion of the molecule. The reduction, by 1D7, of HDL cholesteryl ester uptake is not due to apoE acquisition by the labeled HDL preparation or by the transfer of [3H]CE of HDL to apoE-containing lipoproteins and uptake by the apoB/E or apoE receptors. Rather, it appears that mAb 1D7 recognizes apoE localized at the cell surface of HepG2 cells. This conclusion is supported by the fact that: 1) reduction of HDL-CE uptake by HepG2 cells is observed within 15 min after the addition of the antibody-ligand mixture; 2) 1D7 is similarly effective in reducing the selective uptake of HDL-CE when added to the ligand or to the cells; 3) three different anti-apoE mAbs (1D7, 3B7, and 3H1) bind specifically to the surface of the cells. We have also demonstrated that heparin (5 mg/ml) does not reduce the amount of apoE-immunoreactive material bound at the cell surface when added before or after the binding period. 1D7, but not 3B7 or 3H1, binds less in the presence of heparin. The observations are consistent with a localization of apoE on the cell membrane rather than on lipoproteins bound to apoB/E or apoE receptors. PMID- 8420943 TI - Isozyme-specific modules on human aldolase A molecule. Isozyme group-specific sequences 1 and 4 are required for showing characteristics as aldolase A. AB - Vertebrate aldolase molecules bear at least four stretches of isozyme group specific sequences (referred to as IGS). The IGSs of the type A isozyme are known to endow the aldolase molecules with some characteristics typical of A. In order to locate the type A regions, 4 chimeric enzymes were constructed between human aldolases A and B and 5 mutant enzymes with single or double mutations in the IGS 1 region. Among engineered proteins, the chimeric enzymes bearing the type A IGS 1 to -4 (BABA34-108:306-363) and the IGS-1 and -4 (BABA34-55:306-363) exhibited similarities to isozyme A in many respects. On the other hand, neither chimeric enzyme bearing the type A IGS-1 to -3 (BAB34-108) nor that bearing the IGS-1 alone (BAB34-55) exhibited properties as isozyme A. Four mutant aldolases A (carrying single mutation in the IGS-1 region) maintained the original activity as A. Similarly, the BA306 chimera with the type B-->A substitution at positions 41 and 45 (BA306 N41K:R45S) failed to exhibit the A-like properties although the activities toward Fru-1,6-P2 and Fru-1-P significantly increased. Conclusively, the type A IGS-1, together with the IGS-4, act as indispensable modules in determining the characteristic properties of human aldolase A. PMID- 8420944 TI - Precise location of the Cu(II)-inhibitory binding site in higher plant and bacterial photosynthetic reaction centers as probed by light-induced absorption changes. AB - Light-dependent absorption change at 325 nm, ascribed to QA activity, was strongly reduced in the presence of Cu(II) in oxygen-evolving core complex. This change was much less affected in the presence of the herbicide 3-(3,4 dichlorophenyl)-1,1-dimethylurea (DCMU), indicating that the Cu(II)-binding site is different from that of the DCMU and that Cu(II) blocks QA reduction. Cu(II) did not eliminate the absorption change at 545 nm, ascribed to pheophytin reduction, in Na2S2O4-treated oxygen-evolving core and D1-D2-cytochrome b559 complexes. This indicates that Cu(II) does not affect the electron transport between P680 and pheophytin. Moreover, the activity of the bacterial reaction center probed by the absorption change at 790 nm was inhibited by Cu(II), but the signal at 530 nm, associated to the reduction of bacteriopheophytin in Na2S2O4 treated reaction center, was not inhibited. We conclude that Cu(II) impaired the photosynthetic electron transport between pheophytin and QA in both higher plants and photosynthetic bacteria. Cu(II) would bind to an amino acid(s) highly conserved in non-oxygenic and oxygenic reaction centers, which is(are) necessary for the electron transfer between pheophytin and QA. Based on the atomic structure of the bacterial reaction center several schemes of possible Cu(II) binding are shown. PMID- 8420945 TI - Cooperative phenomena in binding and activation of Bordetella pertussis adenylate cyclase by calmodulin. AB - The catalytic domain of Bordetella pertussis adenylate cyclase located within the first 400 amino acids of the protein can be cleaved by trypsin in two subdomains (T25 and T18) corresponding to ATP-(T25) and calmodulin (CaM)-(T18) binding sites. Reassociation of subdomains by CaM is a cooperative process, which is a unique case among CaM-activated enzymes. To understand better the molecular basis of this phenomenon, we used several approaches such as partial deletions of the adenylate cyclase gene, isolation of peptides of various size, and site-directed mutagenesis experiments. We found that a stretch of 72 amino acid residues overlapping the carboxyl terminus of T25 and the amino terminus of T18 accounts for 90% of the binding energy of adenylate cyclase-CaM complex. The hydrophobic "side" of the helical region situated around Trp242 plays a major role in the interaction of adenylate cyclase with CaM, whereas basic residues that alternate with acidic residues in bacterial enzyme play a much less important role. The amino-terminal half of the catalytic domain of adenylate cyclase contributes only 10% to the binding energy of CaM, whereas the last 130 amino acid residues are not at all involved in binding. However, these segments of adenylate cyclase might affect protein/protein interaction and catalysis by propagating conformational changes to the CaM-binding sequence which is located in the middle of the catalytic domain of bacterial enzyme. PMID- 8420946 TI - Characterization of a synthetic calmodulin-binding peptide derived from Bacillus anthracis adenylate cyclase. AB - A 34-amino acid peptide corresponding to residues 532-565 of Bacillus anthracis adenylate cyclase (P532-565), a calmodulin (CaM)-activated enzyme, was synthesized by solid phase method. Although not homologous to any known CaM binding sequence, P532-565 exhibits molecular features characteristic of this class of peptides: a higher proportion of basic and hydrophobic residues, segregated onto the two faces of the alpha-helical structure. Fluorescence measurements and gel retardation analysis showed that P532-565 binds CaM in a Ca(2+)-dependent manner, with a binding energy that represents 80% of the binding energy of the adenylate cyclase-CaM complex. Circular dichroism analysis showed that P532-565 exists in solution as a mixture of random-coil and alpha-helical structures and that trifluoroethanol increases the relative proportion of alpha helical population. Analysis of proton NMR spectrum in H2O allowed identification of the different amino acid spin systems and complete spectral assignment. The pattern of nuclear Overhauser effect connectivities, intense NN(i,i + 1) and medium range alpha N(i,i + 3) and alpha beta (i,i + 3) indicate the presence of an alpha-helix in the carboxylterminal end (between residues 551 and 563) in fast exchange with extended structures. These data, together with CaM-binding properties of Bordetella pertussis adenylate cyclase, show that despite rather divergent primary structures, the two bacterial enzymes possess similar structural organization of their binding sites for activator protein. PMID- 8420947 TI - Purification and characterization of membrane-bound chitin synthase. AB - The membrane-bound chitin synthase, a key enzyme of chitin biosynthesis, was purified, for the first time to homogeneity as a zymogen form. Digitonin could solubilize the enzyme from microsomal fraction of the filamentous fungus Absidia glauca, with 60-70% of the enzyme activity. The solubilized form of the enzyme was effectively purified by a sequence of chelating Sepharose, concanavalin A Sepharose, and Mono Q column. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the purified enzyme gave a single band with a molecular weight of 30,000. IgG prepared against this 30-kDa species on SDS-polyacrylamide gel electrophoresis immunoprecipitated chitin synthase. The purified enzyme existed as a zymogen, was converted into active form by treatment with trypsin, and the limited digestion with trypsin produced a little smaller polypeptide (28.5 kDa) of which the amino-terminal sequence was identical to the zymogen. The purified enzyme was the glycoprotein and showed a requirement for Mg2+. N Acetylglucosamine stimulated the enzyme activity approximately 5-fold and polyoxin D, an analogue of substrate, and UDP, a byproduct of enzyme reaction, strongly inhibited the enzyme activity. PMID- 8420948 TI - Mechanism of reaction of fatty acid hydroperoxides with soybean peroxygenase. AB - 13(S)-Hydroperoxyoctadeca-9(Z),11(E),15(Z)-trienoic acid (13-HPOT) was used to probe the mechanism of the hydroperoxide O-O bond cleavage catalyzed by solubilized and partially purified soybean peroxygenase. When reacted with this ferrihemoprotein, it was converted to 13(S)-hydroxyoctadeca-9(Z),11(E), 15(Z) trienoic acid (13-HOT) and a single epoxide regio-isomer, i.e. 15,16-cis-epoxy 13(S)-hydroxyoctadeca-9(Z),11(E)-dienoic acid (15,16-EHOD). In the absence of co oxidizable substrates, such as oleic acid or thiobenzamide, this latter compound accounted for about two-thirds of the reaction products. 13-HOT and 15,16-EHOD are products of heterolytic scission of the O-O bond of 13-HPOT; no products arising by homolytic scission could be detected. Therefore, soybean peroxygenase catalyzes hydroperoxide reduction exclusively by a heterolytic mechanism leading to a ferryl-oxo complex analogous to peroxidase compound I. In similar experiments, 13(S)-hydroperoxyoctadeca-9(Z),11(E)-dienoic acid gave 13(S) hydroxyoctadeca-9(Z),11(E)-dienoic acid and 9,10 epoxy-13(S)-hydroxyoctadec-11(E) enoic acid. Experiments with 18O-labeled 13-HPOT indicated that about 83% of the oxygen atom incorporated into the epoxide group of 15,16-EHOD, originated from the hydroperoxide group. Moreover, using mixtures of unlabeled and 18O-labeled 13 HPOT, it was established that this transfer takes place predominantly (about 3:1) by an intramolecular process. In the intermolecular reaction 13-HOT, formed after reduction of the hydroperoxide, diffuses from the active site and, after reassociation, is epoxidized at the 15,16-double bond. A unifying mechanistic scheme, which takes into account all of the reactions catalyzed by the peroxygenase, is proposed. PMID- 8420949 TI - Influenza hemagglutinin-mediated membrane fusion does not involve inverted phase lipid intermediates. AB - Intermediate lipid structures such as inverted micelles and interlamellar attachments, which can form near liquid crystalline lamellar (L alpha) to inverted hexagonal (HII) phase boundaries, are thought to play a role in membrane fusion. To investigate whether these structures are also involved in influenza hemagglutinin-mediated membrane fusion, measurement of fusion under conditions where such structures could not form was attempted. It was found that the fusion of influenza virus with liposomal membranes containing phosphatidylcholine and gangliosides, which cannot form HII phases, was only slightly slower than fusion with liposomes that also contained the HII competent phosphatidylethanolamine. Furthermore, the virus fused efficiently with liposomes consisting either of pure saturated phosphatidylcholines or phosphatidylcholine/ganglioside mixtures, even when the liposomal membranes were in the gel (L beta') phase and thus far from L alpha/HII transitions. Isolated hemagglutinin, reconstituted into dimyristoylphosphatidylcholine membranes, induced fusion with liposomes composed of dimyristoylphosphatidylcholine and gangliosides at temperatures below the L beta' to L alpha phase transition temperature of dimyristoylphosphatidylcholine. This latter finding excluded the possibility that the viral lipids alone could have formed inverted phase intermediates, thus enabling them to fuse with liposomes that do not contain lipids capable of forming inverted phases. Therefore, it is concluded that structures resembling intermediates in L alpha/HII transitions are most likely not involved in influenza hemagglutinin mediated fusion. PMID- 8420950 TI - A luteinizing hormone receptor with a severely truncated cytoplasmic tail (LHR ct628) desensitizes to the same degree as the full-length receptor. AB - The wild type murine luteinizing hormone (mLH) receptor, which in its mature form is predicted to be a protein of 674 amino acids (mLHR), and an artificially mutated form lacking the last 46 amino acids (mLHR-ct628) were stably expressed in murine L cells. Both forms stimulated adenylyl cyclase and underwent rapid desensitization. The mutation removed 1 tyrosine, 2 threonines, and 6 serines from the receptor. The results indicate that none of these potential phosphorylation sites participates in either adenylyl cyclase stimulation or receptor desensitization. Our results with the mLHR-ct628 (carboxyl-terminal amino acid sequence CCKHRAEL) differ from those reported recently for the essentially identically mutated rat LHR that lacks the last 43 amino acids (rLHR ct631 with carboxyl-terminal amino acid composition CCKRRAELYRR). This 43-amino acid truncation was described to have the effect of preventing hormone-induced desensitization. While the reasons for the discrepant results are not known, our results do not support the proposal for a participatory role of the extreme carboxyl terminus of the receptor in its desensitization. PMID- 8420951 TI - Ca(2+)-dependent and Ca(2+)-independent isozymes of protein kinase C mediate exocytosis in antigen-stimulated rat basophilic RBL-2H3 cells. Reconstitution of secretory responses with Ca2+ and purified isozymes in washed permeabilized cells. AB - Rat basophilic RBL-2H3 cells, which exhibit Ca(2+)-dependent secretion of granules when stimulated with antigen, contained the Ca(2+)-dependent alpha and beta and the Ca(2+)-independent delta, epsilon, and zeta isoforms of protein kinase C. These isoforms associated, to variable extents (i.e. delta the most and zeta the least), with the membrane fraction upon antigen stimulation but without external Ca2+; only the Ca(2+)-independent isoforms did so. Both types of isozymes were probably necessary for optimal responses to antigen as indicated by the following observations. All Ca(2+)-dependent isozymes were degraded in cells treated with 20 nM phorbol 12-myristate 13-acetate for 6 h, whereas the Ca(2+) independent isozymes were not degraded and were retained when the cells were subsequently permeabilized and washed. Cells so treated still exhibited antigen induced secretion (25-33% of normal) which was suppressed by selective inhibitors of protein kinase C (Ro31-7549 and calphostin C) thereby indicating a possible contribution of the Ca(2+)-independent isozymes in secretion. Normally, washed permeabilized cells lost all isozymes of protein kinase C and failed to secrete in response to antigen. A full secretory response to antigen could be reconstituted by the subsequent addition of nanomolar concentrations of either beta or delta isozymes of protein kinase C (other isozymes were much less effective) but only in the presence of 1 microM free Ca2+ to indicate distinct roles for Ca2+ and protein kinase C in exocytosis. PMID- 8420952 TI - An N-terminal glycosylation signal on cytochrome P450 is restricted to the endoplasmic reticulum in a luminal orientation. AB - The mechanism of retention of cytochrome P450 in the endoplasmic reticulum is unknown, and the membrane topology of the N-terminal region remains controversial. To address these problems, a sequence of 29 amino acids encoding an internal N-glycosylation site of rabbit cytochrome P450 2C2 was attached to the N terminus of cytochrome P450 2C1. This protein is glycosylated at a single site in a cell-free translation system containing microsomal membranes, as indicated by gel mobility and sensitivity to endoglycosidase H. When expressed in COS1 cells, an immunoreactive species with the same gel mobility as the in vitro synthesized glycosylated product was detected. Treatment with endoglycosidase H changed its mobility to that of unglycosylated hybrid cytochrome P450 2C1. These results indicate that in intact cells, as in the cell-free system, the N terminus of cytochrome P450 is luminally oriented which is not consistent with a hairpin loop conformation. Sensitivity of the glycosylated protein to endoglycosidase H suggests that the protein does not reach the Golgi compartments. When transfected cells were incubated at low temperatures to inhibit retrograde transport from the intermediate pre-Golgi compartment into the endoplasmic reticulum, localization of cytochrome P450 was not changed, as assayed by subcellular fractionation and immunofluorescent staining. These observations suggest that cytochrome P450 is restricted to the endoplasmic reticulum membrane by a mechanism different from recycling through the intermediate compartment, which is a pathway utilized by soluble endoplasmic reticulum proteins. PMID- 8420953 TI - Insulinomimetic effect on glucose transport by epidermal growth factor when combined with a major histocompatibility complex class I-derived peptide. AB - Peptides derived from the alpha 1-region of the murine H-2Dk molecule enhance glucose uptake in rat adipose cells above the maximum obtained with insulin stimulation alone (Stagsted, J., Reaven, G. M., Hansen, T., Goldstein, A., and Olsson, L. (1990) Cell 62, 297-307). We now describe that epidermal growth factor (EGF) in combination with the same peptides, Dk-(61-85) and Dk-(62-85), stimulates cellular glucose uptake 5-7 times over the basal level, i.e. to 30-50% of the maximal insulin effect. EGF alone increased glucose uptake by only approximately 50% above basal and the peptide alone by 100% above basal. Maximal effect of EGF and peptide was reached in 10-20 min with 30 microM peptide (EC50 10-15 microM) and 50 nM EGF (EC50 1-2 nM). The effect of EGF and peptide on glucose uptake was additive to that of insulin and peptide until the maximal level attained with insulin and peptide was reached. The combined effect of EGF plus peptide on glucose transport was associated with a recruitment of GLUT4 molecules to the plasma membrane. However, the phosphatidylinositol (PI) kinase which is activated by insulin was not activated by EGF plus peptide. Thus, the effect of EGF plus peptide on glucose uptake seems independent of the activity status of the insulin receptor. 125I-Labeled EGF bound specifically to rat adipose cells with an apparent affinity of approximately 2 nM and Bmax approximately 5 x 10(3). However, the major histocompatibility complex (MHC) peptides did not affect EGF-stimulated internalization of EGF receptor, in contrast to their effect on the insulin receptors. Transforming growth factor alpha had an effect similar to EGF on glucose uptake. Three other peptides derived from other parts of murine MHC class I had no effect on glucose uptake in combination with EGF. Thus, EGF in combination with certain MHC class I-derived peptides is insulinomimetic concerning glucose transport and this effect is independent of the insulin receptor activity. PMID- 8420954 TI - Differential regulation of adenylylcyclases in vegetative and gametic flagella of Chlamydomonas. AB - To learn more about the mechanism of regulation of cAMP during fertilization in Chlamydomonas, we have begun to study the flagellar adenylylcyclase. Recently (Zhang, Y., Ross, E. M., and Snell, W. J. (1991) J. Biol. Chem. 266, 22954-22959) we reported that the adenylylcyclase in gametic flagella is inhibited by ATP and activated by pretreatment at 45 degrees C or by incubation with the protein kinase inhibitor staurosporine. Here we present evidence that this novel regulatory mechanism is unique to gametes and may be required for sexual signaling between mt+ and mt- gametes during fertilization. The vegetative form of the enzyme, which has a specific activity 3-5-fold less than the gametic form, was neither inhibited by ATP nor activated at 45 degrees C. 5' Adenylylimidodiphosphate, staurosporine, and Mn2+, which activated the gametic enzyme, had no effect on the vegetative adenylylcyclase. In addition the gametic enzyme was inhibited by low (0.1-1 microM) Ca2+ concentrations, whereas the vegetative form was unaffected by 10 microM Ca2+. During gametogenesis acquisition of the ability to undergo sexual signaling was coincident with the appearance of the gametic flagellar adenylylcyclase. Our results suggest that gametogenesis is accompanied by appearance of a new adenylylcyclase that may play a central role in signal transduction during fertilization in Chlamydomonas. PMID- 8420955 TI - Binding of DNA quenches tyrosine fluorescence of RecA without energy transfer to DNA bases. AB - The binding of single- as well as double-stranded DNA to RecA, in the presence of the cofactor analog ATP gamma S (adenosine 5'-O-(3-thiotriphosphate)), leads to about 20% quenching of the tyrosine fluorescence of the protein but to no essential change of the tryptophan fluorescence. The excitation spectrum of the fluorescent DNA analog poly(d epsilon A), complexed with RecA, shows no sign of energy transfer from the tyrosine residues of RecA to the etheno-modified adenine bases of the polynucleotide. From this observation we reject stacking interaction between tyrosine residues and DNA bases. The RecA filament may bind up to three molecules of single-stranded DNA; however, the observed fluorescence change occurs only upon the binding of the first DNA strand, indicating that the binding mode of this first strand is different from those of the others. The fluorescence change is interpreted in terms of a conformational change of the RecA protein promoted by cooperative binding to DNA. A larger quenching (40%) upon the binding of single-stranded DNA is observed in the absence of cofactor. At high salt condition, which induces ATPase activity in RecA just as DNA binding does, the tyrosine fluorescence is more pronounced than at low salt conditions, indicating that the effect induced by high salt is different from the conformational change induced by DNA binding. PMID- 8420956 TI - Role of tyrosine residue 264 of RecA for the binding of cofactor and DNA. AB - The tyrosine fluorescence of the RecA protein is quenched by about 15% upon binding of the cofactor analog adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S). This quenching is not observed with a modified RecA in which the tyrosine residue at position 264 (Tyr-264) is replaced for alanine by site-directed mutagenesis, a modification which also results in a decrease of binding affinity of cofactor. This indicates that Tyr-264 is responsible for the fluorescence change and that the residue is close to or within the cofactor binding site. Upon DNA binding, a change of tyrosine fluorescence is observed both with the modified protein and with wild type RecA, indicating that DNA binding affects the environment of other tyrosine residues than Tyr-264. However, the change is significantly smaller in the modified protein, suggesting that both Tyr-264 as well as other residue(s) may be affected by the DNA binding. Changed fluorescence properties of the remaining tyrosine residues as a result of a slightly different DNA binding mode of the modified protein are also possible. Tyr-264 may be an important residue for the allosteric effect induced by the cofactor for the binding of DNA to RecA. In the recent crystal structure of RecA-ADP published by Story and Steitz (Story, R.M., and Steitz, T. A. (1992) Nature 355, 374-376), ADP is stacked with Tyr-103 and does not interact with Tyr-264. The fact that we observe no interaction of ATP gamma S with Tyr-103 (as evidenced from absence of fluorescence change) but instead with Tyr-264 may suggest an important conformational difference between the RecA complexes with, respectively, ADP and ATP. PMID- 8420957 TI - Cloning and sequencing of a cDNA encoding Saccharomyces cerevisiae carnitine acetyltransferase. Use of the cDNA in gene disruption studies. AB - cDNA encoding for carnitine acetyltransferase (CAT) of yeast S. cerevisiae was isolated by screening a yeast cDNA lambda gt11 library with antibody. The whole coding sequence was obtained from the cDNA and from a YEP 13 DNA clone identified using the cDNA as probe. The coding sequence consists of 670 residues, which amounts to a molecular mass of 77,300 kDa. This cDNA was used successfully to disrupt the gene for the mitochondrial isoenzyme of CAT, which was shown by measuring the enzyme activity and by immunoblot. The acetylcarnitine content of these cells decreased significantly. A search in the PIR protein data base revealed that besides the known carnitine acyltransferases, choline acyltransferases are highly homologous to yeast CAT. The mitochondrial CAT deficient (CAT-) cells were able to grow on different fermentable and nonfermentable carbon sources, even on acetate at the same rate as the parental strain. In contrast to these, 13C NMR studies revealed significant differences between parental and CAT- cells. In CAT-cells [3-13C]pyruvate was converted mainly to lactate and acetate, whereas in the parental cells alanine and tricarboxylic acid cycle intermediates were found as the main products of pyruvate metabolism beside acetate. These results suggest diminished flux through the pyruvate dehydrogenase complex in the absence of mitochondrial CAT in yeast cells. PMID- 8420958 TI - Cold adaptations in Drosophila. Qualitative changes of triacylglycerols with relation to overwintering. AB - Triacylglycerols are the major fuel for basal metabolism during the winter in temperate species of the Drosophila melanogaster species group. Differential scanning calorimetry analysis revealed that the transition temperatures of triacylglycerols were lower in diapausing adults than in reproducing ones, and also lower in species or strains adapted to cooler climates than those adapted to warmer climates. These phenomena were correlated to the fatty acid compositions of the triacylglycerols; the proportion of unsaturated fatty acids in the triacylglycerols was higher in the diapausing individuals, and in the species or strains adapted to cooler climates. Furthermore, in the temperate species of the montium species subgroup (D. subauraria, D. biauraria, D. triauraria, and D. rufa), the amount of saturated triacylglycerols was smaller than the value expected on the assumption that fatty acids are randomly distributed in the triacylglycerols, suggesting the nonrandom distribution of unsaturated fatty acids among triacylglycerols. This may facilitate the lowering of the transition temperature of triacylglycerols, and hence may be related to the ability of Drosophila to cope with temperate climates. PMID- 8420959 TI - Cloning and functional expression in yeast of two human isoforms of the outer mitochondrial membrane channel, the voltage-dependent anion channel. AB - The voltage-dependent anion channel (VDAC) of the outer mitochondrial membrane is a small abundant protein found in all eukaryotic kingdoms which forms a voltage gated pore when incorporated into planar lipid bilayers. VDAC is also the site of binding of the metabolic enzymes hexokinase and glycerol kinase to the mitochondrion in what may be a significant metabolic regulatory interaction. Recently, there has been speculation that there may be multiple forms of VDAC in mammals which differ in their localization in the outer mitochondrial membrane and in their physiological function. In this report, we describe the identification and characterization of two human cDNAs encoding VDAC homologs (HVDAC1 and HVDAC2). To confirm VDAC function, each human protein has been expressed in yeast lacking the endogenous VDAC gene. Human proteins isolated from yeast mitochondria formed channels with the characteristics expected of VDAC when incorporated into planar lipid bilayers. In addition, expression of the human proteins in such strains can complement phenotypic defects associated with elimination of the endogenous yeast VDAC gene. Since VDAC is the site of binding of hexokinase to the outer mitochondrial membrane, the binding capacity of each VDAC isoform expressed in yeast mitochondria was assessed. When compared with the binding of hexokinase to mitochondria lacking VDAC, the results show that mitochondria expressing HVDAC1 are capable of specifically binding hexokinase, whereas mitochondria expressing HVDAC2 only bind hexokinase at background levels. The expression of each human cDNA has been assessed by Northern blot and polymerase chain reaction techniques. With one exception, each is expressed in all human cell lines and tissues examined. PMID- 8420960 TI - Polypeptide composition of the alpha-latrotoxin receptor. High affinity binding protein consists of a family of related high molecular weight polypeptides complexed to a low molecular weight protein. AB - alpha-Latrotoxin is a vertebrate neurotoxin from black widow spider venom that causes massive neurotransmitter release. In order to gain insight into the mechanism of action of alpha-latrotoxin, we have studied alpha-latrotoxin-binding proteins from bovine and rat brain. Proteins purified by affinity chromatography on immobilized alpha-latrotoxin were investigated. Two sets of proteins were isolated: 1) three polypeptides of M(r) 79,000, 65,000, and 43,000 that were eluted from immobilized alpha-latrotoxin by increasing KCl concentrations in the presence of Ca2+, and 2) a family of related proteins ranging in molecular weight from 160,000 to 220,000 and a low molecular weight component of M(r) 29,000 that were eluted from immobilized alpha-latrotoxin only after removal of Ca2+. Amino acid sequences of these proteins demonstrated that all of these proteins represent novel proteins except for the M(r) 65,000 polypeptide, which is identical with synaptotagmin (Petrenko, A. G., Perin, M. S., Davletov, B. A., Ushkaryov, Y. A., Geppert, M., and Sudhof, T. C. (1991) Nature 353, 65-68). Surprisingly, the M(r) 79,000 and 43,000 proteins were also found in tissues insensitive to alpha-latrotoxin action. Since these proteins do not bind 125I alpha-latrotoxin with high affinity, their purification probably is not physiologically significant. On the other hand, the fractions containing the M(r) 160,000-220,000 and 29,000 polypeptides bound alpha-latrotoxin with high affinity. Sucrose gradient centrifugations and anion exchange chromatography suggested that most of the M(r) 160,000-220,000 proteins were complexed with the M(r) 29,000 protein. alpha-Latrotoxin binding correlated with the presence of the M(r) 160,000-220,000 proteins and M(r) 29,000 polypeptide, and alpha-latrotoxin formed stable complexes with the M(r) 160,000-220,000 proteins. Accordingly, the alpha-latrotoxin receptor consists of a high molecular weight protein (M(r) 160,000-220,000) that is complexed with one or several copies of an M(r) 29,000 polypeptide. In addition, the receptor is found in a less tight association with synaptotagmin but not with other polypeptides. PMID- 8420961 TI - Translational regulation of mitochondrial differentiation in neonatal rat liver. Specific increase in the translational efficiency of the nuclear-encoded mitochondrial beta-F1-ATPase mRNA. AB - Postnatal (1-h) mitochondrial differentiation in normal neonatal rat liver is regulated at the translational level (Izquierdo, J. M., Luis, A. M., and Cuezva, J. M. (1990) J. Biol. Chem. 265, 9090-9097). The rapid postnatal increase in liver global rate of protein synthesis preferentially affects mitochondrial proteins (Valcarce, C., Navarrete, R. M., Encabo, P., Loeches, E., Satrustegui, J., and Cuezva, J. M. (1988) J. Biol. Chem. 263, 7767-7775). Analysis of polysome profiles and determination of both eukaryotic initiation factor 2 (eIF-2) activity and amount of eIF-2 beta protein in the liver of fetal and 1-h-old neonatal rats, indicate a rapid activation of translation initiation without changes in the amount of the translational machinery available between both stages of liver development. Appearance of a more acidic eIF-2 beta-subunit form in two-dimensional Western blots from 1-h-old rat livers suggests that covalently regulated modifications of the initiation factor phosphoproteins might be responsible for increased translation in the neonatal liver. On the other hand, preferential cytosolic translation of the mitochondrial nuclear-encoded beta-F1 ATPase mRNA at this stage of liver development is accomplished by (i) the antenatal accumulation of this mRNA in the fetal liver in 5-6-fold excess than that found in adults, although fetal liver beta-F1-ATPase mRNA shows negligible translational efficiency when compared to the adult counterpart; (ii) a 2-fold increase of the stored beta-F1-ATPase mRNA being rapidly mobilized into cytosolic polyribosomes, and (iii) a 3-fold increase in the in vitro determined translational efficiency of beta-F1-ATPase mRNA. Increased translational efficiency of beta-F1-ATPase mRNA at 1-h postnatal is specific for the nuclear encoded template since beta-tubulin mRNA did not show any postnatal alteration in its translational efficiency. The results presented suggest that developmental changes in the poly(A)+ RNA fraction or in the reporter template itself are responsible for the increased and preferential translation of the nuclear-encoded mitochondrial mRNAs needed for mitochondrial differentiation and, thus, for mammalian adaptation to the extrauterine environment. PMID- 8420962 TI - Control of protein traffic between distinct plasma membrane domains. Requirement for a novel 108,000 protein in the fusion of transcytotic vesicles with the apical plasma membrane. AB - We have developed a cell-free system that reconstitutes the last step in transcytosis, i.e. the fusion of transcytotic transport vesicles with the apical plasma membrane (PM). Subcellular fractions containing transcytotic vesicles (the donor) or apical PM (the acceptor) were prepared from rat liver by sucrose density centrifugation. Fusion between the donor and acceptor fractions was measured by the conversion of the 120,000 transmembrane form of the polymeric IgA receptor (pIgA-R), an endogenous protein of transcytotic vesicles, to a processed fragment by a protease endogenous to the apical PM. Fusion occurred only at 37 degrees C and was critically dependent on the presence of ATP and cytosol. Fusion was inhibited by treating the in vitro fusion reaction with N-ethylmaleimide or by adding antibodies against N-ethylmaleimide-sensitive factor (NSF). We have previously identified a specific transcytotic vesicle-associated protein (TAP) and here show that TAP exists in both cytosolic and membrane-associated pools. Because of its exclusive interaction with transcytotic vesicles, we tested the involvement of TAP in distinct fusion processes. Removal of TAP inhibited fusion in an in vitro transcytotic fusion reaction but had no inhibitory effect in an in vitro endosome-endosome fusion system or in an in vitro intra-Golgi transport reaction. We propose that TAP represents part of the molecular machinery specifically involved in targeting and/or fusion of transcytotic vesicles with the apical PM. PMID- 8420963 TI - Cloning of a novel surface antigen from the insect stages of Trypanosoma brucei by expression in COS cells. AB - Trypanosoma brucei cDNA libraries constructed in the vector pCDM8 were screened selectively for insect (procyclic) stage surface antigen cDNAs by transient expression in mammalian COS-7 fibroblasts and "panning" with a rabbit polyclonal antiserum. This strategy yielded two surface antigen cDNAs termed PSSA-1 and PSSA 2. The PSSA-1 cDNA encoded an isotype of procyclin, the major phosphatidylinositol-linked stage-specific glycoprotein antigen of the tsetse fly infective forms of T. brucei. The PSSA-2 cDNA encoded a new and previously unidentified stage-specific surface antigen with the features of a typical transmembrane glycoprotein but with an unusual cytoplasmic tail composed of a proline-rich tandem repeat. Fluorescent antibody staining of PSSA-1 transfected COS cells with a panel of procyclin-specific monoclonal antibodies confirmed that the protein was located on the outer surface of the plasma membrane. Furthermore, the antigen on COS cells was insensitive to treatment with phosphatidylinositol specific phospholipase C suggesting that the inositol of the glycosylinositol phospholipid-lipid anchor contained the same fatty acyl modification reported recently for the procyclin molecule in cultured procyclic trypanosomes. In contrast the PSSA-2 antigen on COS cells was stained very weakly by whole parasite antisera. Northern blot hybridization revealed that the PSSA-2 antigen was encoded by a single 1.7-kilobase transcript which was present in parasites from the insect procyclic stage of the life cycle but not from the animal bloodstream stage. Southern blot hybridization analysis of DNA from procyclic stage trypanosomes indicated that the gene for PSSA-2 may be present in more than one copy in procyclic trypanosomes. PMID- 8420964 TI - ATP induces a conformational change of the 90-kDa heat shock protein (hsp90). AB - The 90-kDa heat shock protein (hsp90) is a well conserved, abundant cytosolic protein believed to be a "chaperone" of most steroid receptors. We have recently demonstrated that hsp90 has an ATP-binding site and autophosphorylating activity (Csermely, P., and Kahn, C. R. (1991) J. Biol. Chem. 266, 4943-4950). Circular dichroism analysis of highly purified hsp90 from rat liver shows that ATP induces an increase of beta-pleated sheet content of hsp90. Vanadate, molybdate, and heat treatment at 56 degrees C induce a similar change in the circular dichroism spectrum. Fourier transformed infrared spectroscopy reveals an ATP-induced increase in the interchain interactions of the 90-kDa heat shock protein due to an increase in its beta-pleated sheet content. In further studies we found that ATP: 1) decreases the tryptophan fluorescence of hsp90 by 11.6 +/- 1.9%; 2) increases the hydrophobic character of the protein as determined by its distribution between an aqueous phase and phenyl-Sepharose; and 3) renders hsp90 less susceptible to tryptic digestion. Our results suggest that hsp90 undergoes an "open-->closed" conformational change after the addition of ATP, analogous in many respects to the similar changes of the DnaK protein, the immunoglobulin heavy chain binding protein (BiP/GRP78), and hsp70. The ATP-induced conformational change of hsp90 may be important in regulating its association with steroid receptors and other cellular proteins. PMID- 8420965 TI - Activating and inhibitory mutations in the regulatory domain of CheB, the methylesterase in bacterial chemotaxis. AB - In the chemotaxis system of Escherichia coli, CheB promotes sensory adaptation by interacting with the chemotaxis receptor-transducer proteins to catalyze removal of their gamma-glutamyl methyl ester groups. CheB is comprised of two functional domains; the C-terminal domain contains the methylesterase active site, and the N terminal domain regulates the activity of this active site. The chemotaxis system controls CheB methylesterase activity via a mechanism involving phosphorylation of the CheB regulatory domain by the chemotaxis protein kinase CheA. To further explore the communication between the regulatory and methylesterase domains of CheB, I generated mutations in the CheB regulatory domain that affect methylesterase activity in vitro. Three of these mutations (D11K, E58K, and E91K) caused increased methylesterase activity in the absence of phosphorylation, and several other mutations (R42H, R73H, and K107R) caused decreased methylesterase activity in the purified proteins. Several of these mutations (D10N, D11K, R42H, E58K, and K107R) also affected the phosphorylation biochemistry of CheB by reducing the rate of CheA-mediated phosphorylation of CheB and/or by decreasing the autodephosphorylation rate of CheB. In addition, all of these mutations diminished the ability of excess CheA to inhibit CheB methylesterase activity. The locations of these mutations in the deduced three-dimensional structure of the CheB N-terminal domain indicate that the region of the protein surrounding the putative phosphorylation site plays important roles in its interaction with the CheB C-terminal domain as well as in its interactions with CheA. PMID- 8420966 TI - DNA repair by eukaryotic nucleotide excision nuclease. Removal of thymine dimer and psoralen monoadduct by HeLa cell-free extract and of thymine dimer by Xenopus laevis oocytes. AB - Using a human cell-free extract, we have recently shown that thymine dimers are removed from DNA in oligonucleotides 27-29 nucleotides in length (Huang, J. C., Svoboda, D. L., Reardon, J. T., and Sancar, A. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 3664-3668). In this study we find that the excision reaction is dependent on ATP, the excised fragments range in length from 27-32 nucleotides, and have 5'-P and 3'-OH termini. We also found that a thymine-psoralen furan side monoadduct is excised from the DNA with a similar incision pattern, indicating that in humans bulky adducts are removed from DNA by the same enzyme system which hydrolyzes mainly the 22-24th and the 5th phosphodiester bonds 5' and 3', respectively, to the lesion. This incision pattern might be common to eukaryotic excision nucleases as thymine dimers were removed from DNA by the same dual incision pattern by Xenopus laevis oocytes. PMID- 8420967 TI - Sequence analysis of lens beta-crystallins suggests involvement of calpain in cataract formation. AB - Abnormal activation of the protease calpain in the lens may be a cause of cataracts. Cataracts were induced in 10-day-old rats by a single overdose of sodium selenite. The water-insoluble protein from the opaque lens nucleus was separated by two-dimensional electrophoresis, electroblotted onto membranes, and the NH2-terminal sequence of partially degraded beta-crystallin polypeptides determined. Selenite cataractous lenses contained four major structural proteins, beta B1, beta B3, beta A3/A1, and beta A4 crystallins, missing from 5 to 49 amino acids from their NH2 termini. Incubation of intact beta-crystallins with calpain II in vitro produced identical cleavage sites. This provided further evidence for the role of calpain in the production of light scattering insoluble protein in cataractous lenses and also suggested that a similar process may lead to lens protein insolubilization during aging. PMID- 8420968 TI - The motif Tyr-X-X-hydrophobic residue mediates lysosomal membrane targeting of lysosome-associated membrane protein 1. AB - We have investigated the mechanism by which LAMP-1, a principal protein of the lysosomal membrane, is targeted to lysosomes. Mutagenesis and transfection experiments indicate that the motif Tyr-X-X-hydrophobic residue at the carboxyl terminus of the 11-amino acid cytoplasmic tail of the protein constitutes the lysosomal targeting signal for LAMP-1. This motif directs CD44, a cell surface hyaluronate receptor, to the lysosomal membrane, but only when the signal is placed at the carboxyl-terminus of a truncated cytoplasmic tail. The signal did not confer lysosomal targeting when it was situated internally or at the carboxyl terminus of the normal CD44 cytoplasmic tail. An apparent paradox is that similar Tyr-containing sequences mediate internalization, but not lysosomal targeting, of several receptors. Of possible relevance is the additional finding that purified LAMP-1 protein lacks the two carboxyl-terminal residues predicted by cDNA, both of which are essential for proper trafficking. A model is proposed in which lysosomal targeting is distinguished from receptor internalization through proteolytic modification of the internalization signal. PMID- 8420969 TI - Binding of the RNA polymerase I transcription complex to its promoter can modify positioning of downstream nucleosomes assembled in vitro. AB - We have studied the reconstitution of chromatin-like structures in vitro, using purified RNA polymerase I transcription complexes and histone octamers. The plasmid construct used in these studies is a pUC8 derivative in which we have inserted an RNA polymerase I core promoter region of Acanthamoeba castellanii upstream of four repeats of the 5 S rDNA nucleosome positioning sequence (208 base pairs) from Lytechinus variegatus. When histone octamers were reconstituted onto the naked DNA template, the expected nucleosome positioning previously observed using tandem repeats of the same 208-base pair fragment was not obtained (as assayed by restriction enzyme digestion mapping of the inserted region of the plasmid). We show that the location of the RNA polymerase I core promoter region with regard to the tandemly repeated 208-base pair positioning sequence is a major determinant in the positioning of the histone octamers. Reconstituting first with the stalled transcription complex excluded octamers from the promoter region and recovered the expected nucleosome positioning downstream on the four repeats of the 5 S positioning sequence. The observed competition between histone octamers and the transcription complex for the promoter region suggests a great similarity with what has been reported from in vitro studies of RNA polymerase II and III transcription systems. We may be looking at a mechanism of regulation of transcription for the RNA polymerase I. PMID- 8420970 TI - Characterization of a DNA polymerase from the hyperthermophile archaea Thermococcus litoralis. Vent DNA polymerase, steady state kinetics, thermal stability, processivity, strand displacement, and exonuclease activities. AB - We have isolated, cloned, and characterized a DNA polymerase from the hyperthermophile archaea Thermococcus litoralis, the Tli DNA polymerase (also referred to as Vent DNA polymerase). The enzyme is extremely thermostable, having a half-life of 8 h at 95 degrees C and about 2 h at 100 degrees C. Pseudo-first order kinetics at 70 degrees C reveal an extremely low Km for a primed M13mp18 substrate (0.1 nM), coupled with a relatively high Km for dNTPs (50 microM). Accompanying extension rates are on the order of 1000 nucleotides/min. Synthesis by the polymerase is largely distributive, adding an average of 7 nucleotides/initiation event. This distributive synthesis can generate products of at least 10,000 bases. Tli DNA polymerase contains a 3'-->5' exonuclease activity that enhances the fidelity of replication by the enzyme (Mattila, P., Korpela, J., Tenkanen, T. and Pitkanen, K. (1991) Nucleic Acids Res. 19, 4967 4973). A 2-amino acid substitution within the conserved exonuclease domain abolishes both double and single strand-dependent exonuclease activity, without altering kinetic parameters for polymerization on a primed single-stranded template. Strand displacement activity by the mutated and unmutated forms increases with increasing temperature and is enhanced in the exonuclease deficient form of the enzyme. PMID- 8420971 TI - A 40-kDa epidermal growth factor/transforming growth factor alpha-binding domain produced by limited proteolysis of the extracellular domain of the epidermal growth factor receptor. AB - Elucidation of the three-dimensional structure of the complex of the epidermal growth factor (EGF) and its receptor is essential for understanding the molecular mechanisms of the EGF-receptor interaction and EGF-induced receptor-receptor interaction. NMR is useful to investigate interactions in solution between macromolecules at atomic resolution, but has a limitation in molecular masses of target proteins: less than 300 residues. We have prepared a fragment with apparent molecular mass of 40 kDa in SDS gels from the soluble extracellular domain of the EGF receptor (sEGFR, 619 residues) by sequential limited proteolysis with proteinase K and bromelain. This fragment is a monomeric structural domain consisting of 202 amino acid residues (Cys302-Arg503) and 18 kDa sugar chains, and binds EGF and transforming growth factor-alpha (TGF alpha). This 40-kDa domain has a dissociation constant of about 1 microM for human TGF alpha, which is similar to that of the parental sEGFR. sEGFR oligomerizes in response to EGF and TGF alpha, while the 40-kDa domain does not, suggesting that the sequences other than this domain is required for receptor oligomerization. The 40-kDa ligand-binding domain described in this report is suitable for analysis by various physico-chemical approaches such as NMR. PMID- 8420972 TI - A myristoylated pseudosubstrate peptide, a novel protein kinase C inhibitor. AB - Synthetic peptides corresponding to the pseudosubstrate domains of protein kinase C (PKC) have been used as specific inhibitors of PKC in in vitro assays and permeabilized cell systems. However, their use in vivo was hampered by the impermeability of the plasma membrane for such peptides. Here, we show that N myristoylation of the PKC pseudosubstrate nonapeptide Phe-Ala-Arg-Lys-Gly-Ala-Leu Arg-Gln permits its use as an inhibitor of PKC in intact cells. The myristoylated peptide, myr-psi PKC, inhibits phosphorylation of the myristoylated alanine-rich C kinase substrate protein, as induced by 12-O-tetradecanoyl-phorbol-13-acetate, and the activation of phospholipase D by bradykinin, which strictly depends on PKC. Half-maximal inhibition is obtained at concentrations of 8 and 20 microM, respectively. An N-myristoylated peptide derived from an inhibitor protein of the cAMP-dependent protein kinases, Myr-Gly-Arg-Arg-Asn-Ala-Ile-His-Asp-Ile, was ineffective. These results show that myr-psi PKC is a selective and cell permeable inhibitor of PKC. PMID- 8420973 TI - Angiotensin II regulates parathyroid hormone-related protein expression in cultured rat aortic smooth muscle cells through transcriptional and post transcriptional mechanisms. AB - Parathyroid hormone-related protein (PTHrP), a tumor product responsible for malignancy-associated hypercalcemia, is also produced in many normal tissues, including vascular smooth muscle cells (SMC). As PTHrP exhibits vasodilatory properties, we postulated that other vasoactive agents may control PTHrP gene expression in SMC. Addition of angiotensin II to serum-deprived SMC resulted in a marked induction of PTHrP mRNA by 2 h, with a peak (6-10-fold) at 4-6 h. Angiotensin II effects on PTHrP gene expression were inhibited by saralasin, an angiotensin II receptor antagonist, and blocked by actinomycin D and cycloheximide, suggesting a requirement for gene transcription and protein synthesis. Nuclear run-off assays revealed a 3-fold increase in PTHrP gene transcription 1 h after angiotensin II treatment. Angiotensin II also prolonged PTHrP mRNA half-life by 2-3-fold. Angiotensin-induced PTHrP mRNA is partially dependent on cyclooxygenase products and protein kinase C activation. Other vasoconstrictor substances, including serotonin and bradykinin, also stimulated PTHrP expression, whereas the vasodilator atrial natriuretic peptide did not. Addition of recombinant PTHrP-(1-141) significantly inhibited angiotensin II induced SMC DNA synthesis. PTHrP expression is increased by angiotensin II through transcriptional and post-transcriptional mechanisms. In addition, PTHrP modulates the effect of angiotensin II on SMC proliferation. This suggests that PTHrP acts locally in SMC, possibly to oppose the vasoactive and/or growth promoting effects of vasoconstrictor agents such as angiotensin II. PMID- 8420974 TI - Proteolytic processing of human amyloid beta protein precursor in insect cells. Major carboxyl-terminal fragment is identical to its human counterpart. AB - The predominant component of amyloid plaques of Alzheimer's disease is the amyloid beta protein (A beta), a 39-42-amino-acid peptide derived by proteolysis of a family of precursors known as amyloid precursor proteins (APP). In mammalian brain and in cultured mammalian cells, the release of APP amino-terminal fragments into the extracellular medium occurs by a proteolytic cleavage within the A beta domain, thereby precluding amyloidogenesis. Infection of Sf9 insect cells with baculovirus vectors containing APP cDNAs results in high levels of APP expression. The concomitant release of amino-terminal fragments of APP and the production of carboxyl-terminal, cell-associated cleavage products are observed. Here we demonstrate by direct protein microsequencing that the proteolytic processing of APP in the Sf9 cells generates a prominent carboxyl-terminal species that is identical to that produced in human cells, suggesting that the major pathway for proteolytic processing of APP is conserved among metazoans. PMID- 8420975 TI - Evidence for an extended structure of the T-cell co-receptor CD8 alpha as deduced from the hydrodynamic properties of soluble forms of the extracellular region. AB - We expressed soluble forms of the human T-cell coreceptor CD8 alpha extracellular region, CD8 alpha 161, and the amino-terminal immunoglobulin-like domain, CD8 alpha 114, in Chinese hamster ovary cells and Escherichia coli, respectively. Both molecules were readily purified to homogeneity in milligram amounts and were recognized by a large panel of monoclonal antibodies. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis indicated that approximately 70% of CD8 alpha 161 was secreted as a disulfide-linked dimer, but CD8 alpha 114 was not disulfide-linked. To investigate the structural features of CD8 alpha 161 and CD8 alpha 114 under native conditions, we performed gel filtration and sucrose gradient sedimentation analysis. In spite of being partially or totally noncovalently bound, both recombinant molecules were stably associated homodimers, as no monomers could be detected at a fairly low protein concentration (approximately 1 microM). This suggests that the CD8 alpha amino terminal domain alone strongly contributes to chain association. Determination of the Stokes radius (Rs) and sedimentation coefficient (s20,w) gave results consistent with CD8 alpha 114 having a globular shape and CD8 alpha 161 being an asymmetric molecule. Taking into account the contribution of hydration to the frictional coefficient, we obtained for CD8 alpha 161 an axial ratio of approximately 5, when modeled as a prolate ellipsoid. These results indicate that the elongated structure of CD8 alpha 161 is essentially contributed by the hinge region and help to explain how the CD8 alpha is able to bridge the distance between the T-cell surface and its binding site in the alpha 3 domain of major histocompatibility complex class I molecules on the target cell. PMID- 8420976 TI - The D domain of the thyroid hormone receptor alpha 1 specifies positive and negative transcriptional regulation functions. AB - Four structural domains are characteristic of the members of the thyroid/steroid receptor superfamily. Of these, the A/B and D domains are the least conserved. We have investigated the role of two clusters of positively charged amino acids within the D domain of the thyroid hormone receptor alpha 1 (TR alpha 1). The sequences Lys134-Arg-Lys and Arg188-Arg-Lys, individually or together, were substituted to the neutral residues TIT in three mutants named alpha 1-1, alpha 1 2, and alpha 1-3, respectively. Subcellular localization of transiently transfected wild-type and mutated TRs was monitored by immunostaining, using a TR alpha 1-specific antibody. The wild-type and the alpha 1-2 TRs were detected exclusively in the nucleus, in the presence or absence of thyroid hormone. In contrast, the alpha 1-1 and alpha 1-3 mutants accumulated in both cytoplasm and nucleus, underscoring the importance of the Lys134-Arg-Lys residues for correct nuclear targeting. More importantly, although the mutants had unimpaired DNA- and hormone-binding activities, all three had lost positive and negative transcriptional regulatory functions. Thus, transactivation and repression functions can be entirely dissociated from the other properties of the receptor. In addition, substitution of either one of the positively charged amino acid clusters was sufficient to convert the native TR alpha 1 into a dominant, thyroid hormone-dependent receptor antagonist. These observations, which underline the functional relevance of the D domain for TR alpha 1 function, may also have implications for the autosomal dominant syndrome of generalized resistance to thyroid hormone. PMID- 8420977 TI - Changes in the structure and catalytic activities of the bovine pituitary multicatalytic proteinase complex following dialysis. AB - The multicatalytic proteinase complex (proteasome) contains at least four distinct active sites catalyzing the degradation of selected chromogenic substrates (trypsin-like, chymotrypsin-like, and peptidylglutamyl peptide hydrolyzing activities) and proteins such as beta-casein. Oxidized insulin B chain was recently proposed as a model substrate for protein degradation by the multicatalytic proteinase complex (Dick, L. R., Moomaw, C. R., DeMartino, G. N., and Slaughter, C. A. (1991) Biochemistry 30, 2725-2734). We studied the dialysis induced activation of the hydrolysis of oxidized insulin B chain by this enzyme. Removal of EDTA from purified preparations of bovine pituitary multicatalytic proteinase complex by dialysis against Tris-HCl buffers led to marked changes in the catalytic properties and structure of the enzyme. Dialysis produced a time dependent activation of oxidized insulin B chain hydrolysis with predominant cleavage at the Glu13-Ala14 bond. A new chromogenic assay was developed for measurement of this activity. Activation was accompanied by a virtually total inactivation of the chymotrypsin-like, trypsin-like, and peptidylglutamyl peptide hydrolyzing activities. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a loss of the 24-kDa subunit and the appearance of a new band at 21 kDa. Amino-terminal amino acid analysis established that the 21-kDa band was autolytically derived from the 24-kDa subunit. Evidence for partial dissociation and/or aggregation indicated that autolysis destabilizes the complex. By altering the profile of catalytic activities of the multicatalytic proteinase complex, autolysis may serve as a mechanism for regulation of this macromolecule. PMID- 8420978 TI - Aspartyl residue 10 is essential for ATPase activity of rat hsc70. AB - Three mutants of rat hsc70 were constructed, overexpressed in Escherichia coli, purified, and characterized. First, site-directed mutation was utilized to substitute Asn for Asp-10. The recombinant protein, hsc70(D10N), loses not only its peptide-stimulated ATPase activity but also its basal ATPase activity. The measured dissociation constants of ATP (0.3 microM) and S-peptide (5 microM) for hsc70(D10N), however, are virtually identical to those of hsc70. The intrinsic fluorescence spectra of hsc70(D10N) also remain largely unchanged. Therefore, the overall structure of the hsc70 protein is most likely intact after mutation. Second, the entire C-terminal peptide-binding domain was deleted and the resultant mutant contains only the N-terminal ATPase domain of hsc70. This recombinant protein, Nt-hsc70, is a peptide-independent ATPase. The ATPase activity at 37 degrees C of the Nt-hsc70, 270 pmol/h/micrograms of protein, is comparable to that of maximally peptide-activated hsc70. Third, the Asp-10 of Nt hsc70 was replaced by Asn. Despite that this mutant, Nt-hsc70(D10N), is capable of binding ATP and it loses the capability to hydrolyze ATP. Taken together, these results indicate that aspartyl residue 10 of hsc70 is essential for ATP hydrolysis. Purified hsc70 and its mutants autophosphorylate in vitro at a substoichiometric level. On average, less than 1% of the hsc70 and Nt-hsc70 proteins are phosphorylated. Although the amount of phosphate incorporated into hsc70(D10N) and Nt-hsc70(D10) is reduced, a significant level of phosphorylation can still be achieved in these two site-directed mutants. Hence, autophosphorylation of hsc70 and its mutants is not correlated with their ability to hydrolyze ATP. PMID- 8420979 TI - The rat liver ecto-ATPase is also a canalicular bile acid transport protein. AB - A approximately 110-kDa glycoprotein purified from canalicular vesicles by bile acid affinity chromatography has been identified as the canalicular bile acid transport protein. Internal amino acid sequence and chemical and immunochemical characteristics of this protein were found to be identical to a rat liver canalicular ecto-ATPase. In order to definitively determine whether these were two activities of a single polypeptide, we examined the possibility that transfection of cDNA for the ecto-ATPase would confer bile acid transport characteristics, as well as ecto-ATPase activity, on heterologous cells. The results show that transfection of the ecto-ATPase cDNA conferred on COS cells de novo synthesis of a approximately 110-kDa polypeptide, as immunoprecipitated by antibody to the purified canalicular bile acid transport protein and conferred on COS cells the capacity to pump out [3H]taurocholate with efflux characteristics comparable with those previously determined in canalicular membrane vesicles (Km = 100 microM; Vmax = 200 pmol/mg of protein/20 s). A truncated ecto-ATPase cDNA, missing the cytoplasmic tail, was targeted correctly to the cell surface but did not confer bile acid transport activity on COS cells. The results of this study also show that the canalicular ecto-ATPase/bile acid transport protein is phosphorylated on its cytoplasmic tail and that its phosphorylation is stimulated by activation of protein kinase C and inhibited by inhibitors of protein kinase C activation. Moreover, inhibition of protein kinase C activation by staurosporine completely abrogates bile acid transport but does not affect ATPase activity. This study, therefore, demonstrates that the rat liver canalicular ecto-ATPase is also a bile acid transport protein, that the capacity to pump out bile acid can be conferred on a heterologous cell by DNA-mediated gene transfer, and that phosphorylation within the cytoplasmic tail of the transporter is essential for bile acid efflux activity but not for ATPase activity. PMID- 8420980 TI - Accumulation of 1,2-sn-diradylglycerol with increased membrane-associated protein kinase C may be the mechanism for spontaneous hepatocarcinogenesis in choline deficient rats. AB - Choline deficiency, via deprivation of labile methyl groups, is associated with a greatly increased incidence of hepatocarcinoma in experimental animals. This dietary deficiency also causes fatty liver, because choline is needed for hepatic secretion of lipoproteins. We hypothesized that fatty liver might be associated with the accumulation of 1,2-sn-diradylglycerol and subsequent activation of protein kinase C. Several lines of evidence indicate that cancers might develop secondary to abnormalities in protein kinase C-mediated signal transduction. We observed that rats fed a choline-deficient diet for 1, 6, or 27 weeks had increased hepatic concentrations of 1,2-diradylglycerol. At 1 and 6 weeks, hepatic plasma membrane from choline-deficient rats had increased concentrations of 1,2-sn-diacylglycerol and 1-alkyl, 2-acylglycerol, with the latter accounting for 20-26% of membrane 1,2-sn-diradylglycerol (as compared with only 2-5% in controls). Protein kinase C activity was increased in hepatic plasma membrane at 1 week of choline deficiency. By Western blotting there was an increase in the amount of protein kinase C zeta and a decrease in the amount of protein kinase C delta in liver at 1 week. By 6 weeks of choline deficiency, hepatic plasma membrane and cytosolic protein kinase C (PKC) activities were increased significantly, with increased amounts of hepatic plasma membrane protein kinase C alpha, and delta detected by Western blotting. Glycogen synthase activity in liver was diminished after 1 week of choline deficiency; this enzyme is inhibited by PKC-mediated phosphorylation. We suggest that choline deficiency perturbed PKC mediated transmembrane signaling within liver and that this contributed to the development of hepatic cancer in these animals. PMID- 8420981 TI - Molecular characterization of four pharmacologically distinct gamma-aminobutyric acid transporters in mouse brain [corrected]. AB - Two novel gamma-aminobutyric acid (GABA) transporters, GAT3 and GAT4, were cloned from the mouse neonatal brain cDNA library and expressed in Xenopus oocytes. Sequence analysis indicated they were members of the Na(+)-dependent neurotransmitter transporter family. The GABA uptake activities were measured in cRNA injected Xenopus oocytes. The Km for GABA uptake by GAT3 was 18 microM and by GAT4 was 0.8 microM. GAT3 also transports beta-alanine and taurine with Km of 28 and 540 microM, respectively. Similarly, GAT4 transports beta-alanine with Km of 99 microM and taurine with a Km of 1.4 mM. The newly cloned GABA transporters were compared with two previously cloned GABA transporters, GAT1 and GAT2, in terms of molecular and pharmacological properties. While GAT1 and GAT4 gene expression were neural specific, GAT2 and GAT3 mRNAs were detected in other tissues such as liver and kidney, in which GAT3 mRNA was especially abundant. The expression of GAT3 mRNA in mouse brain is developmentally regulated, and its mRNA is abundant in neonatal brain but not in adult brain. High affinity GABA transporters GAT1 and GAT4 were more sensitive to inhibition by nipecotic acid. Low affinity GABA transporters GAT2 and GAT3 were inhibited most effectively by betaine and beta-alanine, respectively. The differential tissue distribution and distinct pharmacological properties of those four GABA transporters suggest functional specialization in the mechanisms of GABA transmission termination. PMID- 8420982 TI - Site-directed mutagenesis of HIV-1 integrase demonstrates differential effects on integrase functions in vitro. AB - The retroviral integrase (IN) protein is essential for integration of retroviral DNA into the host cell genome. To identify functional domains within the protein and to assess the importance of conserved residues, we performed site-directed mutagenesis of HIV-1 IN and analyzed the mutants in vitro for IN-mediated activities: 3' processing (att site-specific nuclease activity), strand transfer (the joining of att site oligonucleotides to target DNA), disintegration (the reverse of strand transfer), and integration site selection. Changing the conserved residue His-16 either to Cys or to Val in a proposed zinc-finger region had minimal effect on IN activities. Alteration of two highly conserved amino acid residues, Asp-116-->Ile and Glu-152-->Gly, each resulted in complete or nearly complete loss of 3' processing, strand transfer, and disintegration, whereas alteration of another conserved residue, Trp-235-->Glu, had no demonstrable effect on any of the activities in vitro. Two mutants, Asp-64-->Val and Arg-199-->Cys delta, each demonstrated differential effects on IN activities. Asp-64-->Val has no demonstrable strand transfer or disintegration activity yet maintains 3' processing activity at a diminished level. Arg-199-->Cys delta, which lacks part of the carboxyl terminus of IN, has impaired strand transfer activity without loss of disintegration activity. Use of a target site selection assay showed that all of our mutants with strand transfer activity maintain the same integration pattern as wild type IN. We conclude that not all highly conserved IN residues are essential for IN activities in vitro, zinc coordination by the proposed zinc-finger domain may not be required for the activities assayed, alteration of single residues can yield differential effects on IN activities, and target site selection into naked DNA is not necessarily altered by changes in strand transfer activity. PMID- 8420983 TI - Direct association of interleukin-6 with a 130-kDa component of the interleukin-6 receptor system. AB - Affinity cross-linking of membrane bound 125I-interleukin-6 (IL-6) on several cell lines revealed a three-band pattern of IL-6-containing cross-linked complexes with molecular masses of 100, 120, and 150 kDa. To identify the membrane components that were associated with IL-6 in the three complexes, we employed the Denny-Jaffe reagent, a heterobifunctional, cleavable cross-linker that allows the transfer of 125I from the ligand to its receptor. Samples cross linked with Denny-Jaffe reagent were analyzed by two-dimensional SDS polyacrylamide gel electrophoresis in which the cross-linker was cleaved prior to the second dimension. This analysis revealed that IL-6 directly associates with a 130-kDa membrane protein thus allowing the formation of the 150-kDa complex. In addition, both the 100- and 120-kDa cross-linked complexes were shown to include an 80-kDa membrane glycoprotein associated with one and two IL-6 molecules, respectively. PMID- 8420984 TI - Regulation of human tissue factor expression by mRNA turnover. AB - Tissue factor serves as the cellular receptor for circulating blood coagulation factor VII and is the principal physiological initiator of blood coagulation. Tissue factor is not normally expressed in cells that contact blood, such as endothelial cells and monocytes, but can be induced in these cells by tumor necrosis factor or tumor-promoting phorbol esters. Following induction, the human tissue factor mRNA is degraded with a half-life of approximately 0.75-1.5 h. The cellular mechanisms responsible for this rapid mRNA turnover were investigated with chimeric tissue factor.beta-globin constructs expressed in stably transfected mouse NIH/3T3 cells. These constructs were expressed with the transiently inducible c-fos promoter which eliminated the need to use transcriptional inhibitors to determine mRNA half-lives. Sequences capable of conferring rapid turnover to the normally stable beta-globin transcript were localized to the last 600 nucleotides of the tissue factor mRNA. The 3' end of this fragment is similar to previously described AU-rich mRNA destabilizing elements. Activity of the tissue factor element was dependent on its specific sequence and not simply a high AU nucleotide content. The degradation of unstable chimeric tissue factor.beta-globin mRNAs was prevented by inhibition of transcription with actinomycin D. Chimeric tissue factor.beta-globin mRNAs were superinduced by the protein synthesis inhibitor cycloheximide, and this superinduction may be due in part to stabilization of the mRNA. PMID- 8420985 TI - Activation of the human vimentin gene by the Tax human T-cell leukemia virus. I. Mechanisms of regulation by the NF-kappa B transcription factor. AB - The molecular basis for transactivation of the human vimentin gene by the Tax protein from the human T-cell leukemia virus (HTLV-1) is analyzed in this report. We first demonstrate that the factor NF-kappa B binds to the vimentin promoter. Using gel retardation assays, we found the putative NF-kappa B protein. Specific antibodies and competition experiments between the NF-kappa B-binding site in the interleukin-2R alpha and HIV-1 promoters and the vimentin promoter show that the three sites have identical affinity for the factor. We further show that the mechanisms of activation of NF-kappa B by the Tax protein involve a cellular inducer. Nuclear extract from lymphoid cells expressing Tax can induce in vitro a NF-kappa B binding activity in nonlymphoid cytosolic extract. This inducer, if preexisting in an inactivate state in T-cells which are not expressing Tax, cannot be switched to an active state by addition of partially purified Tax protein. While found in the nucleus of Tax-expressing cells in our experiments, this inducer might be cytoplasmic as well. In a first attempt to identify and characterize the inducer, we present the results of fractionation assays of nuclear extract. PMID- 8420986 TI - A pathogen-responsive gene of parsley encodes tyrosine decarboxylase. AB - A group of recently isolated parsley (Petroselinum crispum) cDNAs representing genes that are transcriptionally activated upon fungal infection or elicitor treatment have been demonstrated to encode tyrosine decarboxylase (TyrDC). The deduced TyrDC protein sequence shares extensive similarity with two functionally related enzymes, tryptophan decarboxylase from periwinkle and dopa decarboxylase from Drosophila melanogaster. Expression of TyrDC cDNA in Escherichia coli yielded catalytically active protein with high substrate specificity for tyrosine. All four identified parsley TyrDC genes have been cloned and encode at least three TyrDC isozymes. Treatment of cultured parsley cells with fungal elicitor caused very rapid and transient increases in TyrDC mRNA and enzyme activity levels. PMID- 8420987 TI - Dominant lethal mutations near the 5' substrate binding site affect RNA polymerase propagation. AB - The segment Asp1064-Lys1073 in the beta subunit of Escherichia coli RNA polymerase is evolutionarily conserved and is located near the "5' face" of the nucleotide binding pocket as was shown by affinity labeling with priming substrates (Grachev, M. A., Lukhtamov, E. A., Mustaev, A. A., Zaychikov, E. F., Abdukayumov, M. N., Rabinov, I. V., Richter, V. I., Skoblov, Y. S., and Chistyakov, P. G. (1989) Eur. J. Biochem. 180, 577-585). We engineered single Xaa ->Ala or Ala-->Ser substitutions of eight evolutionarily conserved amino acids in this segment as well as a multiple alanine (KRNK) substitution of four of these residues. The KRNK mutation as well as four of the single substitutions were dominant lethal, two of the single mutations were recessive lethal, and two were viable. RNA polymerase bearing the dominant mutations was prepared for biochemical study by in vitro reconstitution from subunits. All of the mutant enzymes formed stable, specific promoter complexes, capable of initiating RNA synthesis. However, the KRNK polymerase was totally blocked in initiation-to elongation transition, whereas the four point mutants displayed allele-specific changes in promoter clearance rate. Each of the four mutations changed, in a specific way, both the pattern of short oligomers generated in abortive initiation and the pattern of RNA polymerase pausing during elongation. Thus, the mutations appear to distort but not destroy the active center and to alter, in allele-specific manner, the coupling between the catalytic reaction and RNA polymerase propagation along the template. PMID- 8420988 TI - Stimulation of poly(A) tail elongation by the VP39 subunit of the vaccinia virus encoded poly(A) polymerase. AB - The VP55 subunit of the vaccinia virus-encoded poly(A) polymerase can add a maximum of 35 adenylates to the 3'-end of an RNA primer in a rapid and highly processive manner, whereas the VP55-VP39 heterodimer catalyzes the formation of poly(A) tails several hundred nucleotides in length. Here, we describe the overexpression of the VP39 subunit, its purification to near homogeneity, and its ability to associate physically with VP55 and to stimulate polyadenylation. Although VP39 possessed no independent poly(A) polymerase activity, RNA primers with oligo(A) tails greater than 30 adenylates in length could be extended nearly 40-fold more rapidly in the presence of VP39. VP39 enhanced the polyadenylation rate by converting the slow, nonprocessive polyadenylation occurring after the rapid burst in the presence of monomeric VP55, to a rapid, semiprocessive reaction. The effect of VP39 was dramatic when poly(A) primers were used as, 60 mM NaCl, VP39 enhanced the polyadenylation rate 500-fold, and at 90 mM NaCl VP39 was absolutely required. Nevertheless, the VP39-containing polymerase remained selective for polyadenylation of an mRNA 3'-end in the presence of excess poly(A). These data suggest that the role of VP39 in polyadenylation is to increase the affinity of the polymerase for the growing poly(A) tail. PMID- 8420989 TI - Regulatory role of GM3 ganglioside in alpha 5 beta 1 integrin receptor for fibronectin-mediated adhesion of FUA169 cells. AB - Mouse mammary carcinoma mutant cell line FUA169, characterized by high GM3 ganglioside content, was established from parent cell line FM3A/F28-7, which has high lactosyl ceramide (LacCer) content but no GM3. FUA169 displays no changes in protein glycosylation, and is a typical glycolipid mutant differing from its parent in that it contains high quantities of GM3 and GlcCer, but no LacCer (see accompanying paper; Tsuruoka, T., Tsuji, T., Nojiri, H., Holmes, E. H., Hakomori, S. (1993) J. Biol. Chem. 268, 2211-2216). In contrast to parent F28-7 cells, FUA169 cells showed clear adhesion to fibronectin (FN). Several lines of evidence indicate that adhesion of FUA169 cells to FN requires the presence of GM3, which supports the function of integrin receptor. (i) Both FUA169 and F28-7 cells express the same quantity of FN integrin receptor, which consists of alpha 5 beta 1 (sensitive to RGDS peptide) and alpha 4 beta 1 (sensitive to CS1 peptide). However, adhesion to FN-coated plates, regardless of type of FN, was much higher for FUA169 than for F28-7 cells. (ii) F28-7 cells, which normally lack GM3 and adhere only weakly to FN, acquired GM3 during incubation in GM3-containing medium, and subsequently adhered strongly to FN. (iii) Cholesterol-lecithin liposomes (cholesterol was 14C-labeled) incorporating alpha 5 beta 1 receptor isolated from human placenta showed clear adhesion to FN-coated plates, and this adhesion was completely inhibited by RGDS peptide and by anti-beta 1 mAb ZH1. When liposomes included a moderate quantity of GM3 (0.22-0.44 micrograms (0.2-0.4 nmol)/55 micrograms of phosphatidylcholine, 33 micrograms of cholesterol, 5 micrograms of alpha 5 beta 1 in liposome), adhesion was enhanced significantly. In contrast, adhesion was greatly reduced below control level for alpha 5 beta 1 liposomes containing a higher quantity (2.2 micrograms; > 2 nM) of GM3. Adhesion to FN was also inhibited, but never enhanced, for alpha 5 beta 1 liposomes with similar composition but containing 0.4 nmol (or other quantities) of LacCer or GlcCer instead of GM3. These findings suggest that the greater adhesion to FN by FUA169 cells, relative to parent F28-7 cells, is due to functional support by GM3 of alpha 5 beta 1 integrin receptor. PMID- 8420990 TI - Structural and functional changes of lysosomal acid alpha-glucosidase during intracellular transport and maturation. AB - The synthesis and posttranslational modification of lysosomal acid alpha glucosidase were studied in a cell-free translation system and in mammalian cells transfected with acid alpha-glucosidase cDNA constructs. The newly synthesized precursor, sequestered in the endoplasmic reticulum, was demonstrated to be membrane-bound by lack of signal peptide cleavage, and to be catalytically inactive. Sugar chain modification was shown to occur in the Golgi complex and to be dependent on the rate of transport. From the trans-Golgi network different routes were found to be followed by acid alpha-glucosidase. A fraction of precursor molecules, proteolytically released from the membrane anchor, appeared to enter the secretory pathway and was recovered from the cell culture medium in a catalytically active form. A second fraction was transported to the lysosomes and was trimmed in a stepwise process at both the amino- and carboxyl-terminal ends. The intramolecular cleavage sites were determined. Involvement of thiol proteinases was demonstrated. Specificity for the natural substrate glycogen was gained during the maturation process. The phosphomannosyl receptor is assumed to be instrumental in the lysosomal targeting of acid alpha-glucosidase, but a phosphomannosyl receptor-independent transport of membrane-bound precursor molecules to the lysosomes, either directly or via the plasma membrane, cannot be excluded. PMID- 8420991 TI - Dissection of the biotinyl subunit of transcarboxylase into regions essential for activity and assembly. AB - Transcarboxylase, a multisubunit enzyme containing 12 S, 5 S, and 1.3 S subunits, catalyzes the transfer of a carboxyl group from methylmalonyl-CoA to pyruvate (overall reaction) via two partial reactions. In the first partial reaction, a carboxyl group from methylmalonyl-CoA bound to the 12 S subunit is transferred to the biotin of the 1.3 S subunit, and, in the second partial reaction, the carboxylated biotin transfers its carboxyl group from biotin to pyruvate, bound to the 5 S subunit. Previously we have shown that the region around the biotinyl lysine of the 1.3 S subunit is critical for catalysis, that peptides in the amino terminal region of 1.3 S are capable of forming complexes with 12 S and 5 S, and that amino acids in the carboxyl terminus of the 1.3 S subunit form part of the recognition site for holocarboxylase synthetase. In order to further examine the role of the sequences in this subunit, we generated 8 shortened forms of the 1.3 S biotinyl subunits missing either one or both termini. Truncated 1.3 S subunits were active in both partial reactions until deletion reached amino acid 59. None of the truncated subunits was able to support stable complex formation with the 12 S and 5 S subunits or catalyze the overall reaction. The results suggest that the region between 59 and 78 is required for activity and the sequence 1-18 is required for enzyme assembly. Activity in the partial reactions correlated with intrinsic fluorescence enhancement of tryptophan residues in either the 12 S or 5 S subunit. Fluorescence enhancement was observed with the shortened 1.3 S subunits until truncation reached amino acid 59 implying either 1) that the internal sequence, 59-78, transiently associates with the other subunits to properly orient the biotin for catalysis or 2) that the sequence 59-78 contributes to the folded conformation of the 1.3 S subunit so that subunit interactions can take place. PMID- 8420992 TI - Hydrodynamic properties and immunological identification of the sodium- and chloride-coupled glycine transporter. AB - We have recently reported the purification of the native sodium- and chloride coupled glycine transporter from pig brain stem (Lopez-Corcuera, B. Vazquez, J., and Aragon, C. (1991) J. Biol. Chem. 266, 24809-24814). This preparation is essentially homogeneous and contains a unique 100-kDa polypeptide based on electrophoretic migration under denaturing conditions. In this paper we report the hydrodynamic characterization of the native transporter, solubilized in two different detergents, as well as the immunological identification of the protein. On the basis of results obtained from size-exclusion chromatography, we calculated the Stokes radii of transporter 3-[(3-cholamidopropyl)dimethylammonio] 1-propanesulfonic acid (CHAPS) and transporter-cholate complexes to be 5.5 and 6.0 nm, respectively. In addition, from H2O/D2O sucrose density gradient sedimentation analysis, we calculated the molecular weight of protein-detergent complexes to be 115,000 and 160,000 in CHAPS and cholate, respectively, and the molecular weight of the protein moiety as 86,000. Finally, polyclonal antibodies raised against the 100-kDa polypeptide were found to immunoprecipitate specifically glycine transport activity. Taken together, the results reported herein corroborate the identity of the 100-kDa band as the sodium- and chloride coupled glycine transporter and also suggest that in its native state the transporter is a monomeric protein. PMID- 8420993 TI - HRas-dependent pathways can activate morphological and genetic markers of cardiac muscle cell hypertrophy. AB - We have investigated the role of the proto-oncogene HRas in cardiac cell growth and hypertrophy. By direct needle microinjection of activated Ras protein into primary neonatal rat ventricular cardiac myocytes, we find that, unlike many other cell types, Ras does not induce DNA synthesis in these cells. However, injection of activated Ras does induce expression of both the c-Fos and atrial natriuretic factor (ANF) genes. Expression of both these genes is associated with the hypertrophic response in ventricular myocytes suggesting that Ras is involved in the hypertrophic signalling pathway. Ras injection also causes morphological changes in the cells so that they increase in profile and show changes in the organization of the contractile apparatus. Further support for a role for Ras in the hypertrophic response was obtained from studies showing that activated Ras stimulates ANF promoter activity in transient transfection assays. We also show that a dominant interfering Ras mutant inhibits the hypertrophic stimulation of the ANF promoter by phenylephrine, indicating a role for Ras in the hypertrophic effect of an alpha-adrenergic agonist. PMID- 8420994 TI - c-myb transactivates cdc2 expression via Myb binding sites in the 5'-flanking region of the human cdc2 gene. AB - The c-myb protooncogene is preferentially expressed in hematopoietic cells and is required for cell cycle progression at the G1/S boundary. Because c-myb encodes a transcriptional activator that functions via DNA binding, it is likely that c-myb exerts its biological activity by regulating the transcription of genes required for DNA synthesis and cell cycle progression. One such gene, cdc2, encodes a 34 kDa serine-threonine kinase that appears to be required for G1/S transition in normal human T-lymphocytes. To determine whether c-myb is a transcriptional regulator of cdc2 expression, we subcloned a segment of a cdc2 human genomic clone containing extensive 5'-flanking sequences and part of the first exon. Sequence analysis revealed the presence of two closely spaced Myb binding sites that interact with bacterially synthesized Myb protein within a region extending from nucleotides -410 to -392 upstream of the transcription initiation site. A 465-base pair segment of 5'-flanking sequence containing these sites was linked to the CAT gene and had promoter activity in rodent fibroblasts. Cotransfection of this construct with a full-length human c-myb cDNA driven by the early simian virus 40 promoter resulted in a 6-8-fold enhancement of CAT activity that was abrogated by mutations in the Myb binding sites. These data suggest that c-myb participates in the regulation of cell cycle progression by activating the expression of the cdc2 gene. PMID- 8420995 TI - Glycophorin B and glycophorin E genes arose from the glycophorin A ancestral gene via two duplications during primate evolution. AB - Human glycophorin A, B, and E genes are homologous from the 5'-flanking region to 1 kilobase downstream from the exon encoding the transmembrane region. Analysis of human Alu sequences at the transition site from the homologous to nonhomologous region suggested that the GPA gene most closely resembles the ancestral gene, whereas GPB and GPE genes arose by homologous recombination within the Alu repetitive sequence, and acquired 3' sequences from an unrelated gene (Kudo, S., and Fukuda, M. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 4619 4623; Kudo, S., and Fukuda, M. (1990) J. Biol. Chem. 265, 1102-1110). To understand glycophorin gene evolution in primate phylogeny, transmembrane and Alu regions of several primate genomes were amplified by the polymerase chain reaction and their sequences were analyzed. These studies revealed that the GPA gene was present in all primates studied, and the GPB gene was present in pygmy chimpanzee, chimpanzee, and gorilla, but absent from orangutan and gibbon. GPE gene was present in all species with a GPB gene, but was detected in only 7 out of 16 gorillas. The 24-base pair insertion sequence found in the transmembrane exon of the human GPE gene was shown to be derived from the ancestral GPB gene and was inserted into the ancestral GPE gene prior to gorilla divergence. The recombination site in the GPA gene was confirmed to be within an Alu repetitive sequence. We conclude that GPB and GPE genes arose from an ancestral GPA gene via two gene duplications occurring during primate evolution, prior to gorilla divergence. PMID- 8420996 TI - Type I human complement C2 deficiency. A 28-base pair gene deletion causes skipping of exon 6 during RNA splicing. PMID- 8420997 TI - Bacterial adhesion to poly(HEMA)-based hydrogels. AB - The effects of water content and comonomer chemistry upon the adhesion of Pseudomonas aeruginosa to poly(hydroxyethyl methacrylate)-based hydrogels were studied. Hydrogels which varied in swollen water content from 33-69 wt% were polymerized onto glass microscope slides pretreated with a vinyl silane. The hydrogel water content was varied by adding methacrylic acid (1-5 wt%) or N-vinyl pyrrolidone (NVP, 10-25 wt%) or combinations of the two comonomers. The resulting hydrogel surfaces, which were 0.1 mm thick, transparent, and adherent to the glass slide, formed the test surfaces of laminar flow cells (Re = 1.3, wall shear rate = 1.6/s). The bacteria were grown for 8 h in tryptic soy broth (TSB), washed by filtration, and collected on 0.45-microns filters, resuspended in phosphate buffered saline (PBS) at pH = 7.2, and recirculated through the flow cell and across the test surface at 0.85 mL/min for 2 h. Results show that P. aeruginosa adhered less to hydrogels with higher water contents. In the presence of TSB and possible poly(NVP) contamination, the concentration of adherent bacteria was reduced to low and uniform levels independent of the hydrogel chemistry. PMID- 8420998 TI - Kinetic study of collagen fixation with polyepoxy fixatives. AB - A new biomaterial has been developed by fixing native collagen with a polyepoxy compound (PC) fixative. In this study, bovine internal thoracic arteries were fixed with PC under various conditions to help understand the kinetics of the collagen-PC reactions and optimize the fixation process. At predetermined time intervals, small samples were cut from the arteries to determine the quantities of the remaining unreacted amino acids in the collagen. Temperature, concentration, and solution pH were among the key parameters studied. The overall fixation rate was found to be reaction-rate controlled, as the rate of fixation was relatively slow compared with the rate of diffusion of PC. As might be expected, the reaction rate was favored by a higher temperature, concentration, and solution pH. A kinetic model, with a 2.5th reaction order with respect to the reactive functional groups of collagen and a first order with respect to PC, was developed that gave a good fit to the experimental data. Based on this model, the degree of fixation, X, as a function of time, t, is given by (1 - X)-1.5 = 1 + Kt, where K is a constant related to the initial concentrations and the reaction rate constant. PMID- 8420999 TI - Influence of mineral content and composition on graylevels in backscattered electron images of bone. AB - To determine the meaning of graylevels in backscattered electron (BSE) images of actual bone tissues, the influence of mineral content and mineral composition on BSE image graylevels was studied using chick bone tissue representing a broad age range. These tissues were analyzed for BSE image graylevels, Ca/P molar ratios, mineral composition mineral content (v/v), ash fraction (w/w), and density (g/cm3). Linear regression analyses showed that the weighted mean graylevels (WMGLs) in BSE images were positively correlated to ash fraction (r2 = 0.711), mineral content (r2 = 0.720), and density (r2 = 0.843). Although the Ca/P ratio increased from 1.65 in embryos to 1.80 in 2-year olds, the compositional changes corresponding to this Ca/P molar ratio were estimated to produce a relatively minor (< 4.0%) change in BSE image graylevel. These results demonstrate that graylevels in BSE images of actual bone tissue can be attributed to mineral content and density, but only as a coincidence of their association with atomic number. PMID- 8421000 TI - Measurement of intracellular hydrogen peroxide induced by biomaterials implanted in a rodent air pouch. AB - Biomaterials elicit an inflammatory response that is undoubtedly a factor in their healing and in the complications associated with their use. Herein, we report the modification of a rat air pouch in which we measured the production of intracellular hydrogen peroxide by inflammatory cells adherent to the surfaces of silicone elastomer (SE) and expanded polytetrafluoroethylene (ePTFE). We used the fluorescent probe dichlorofluorescein diacetate (DCFH-DA) to measure the intracellular production of hydrogen peroxide 2 and 7 days after implantation. After 2 days, host cells adherent to ePTFE produced almost 3.5 times more hydrogen peroxide than did cells adherent to SE (P < .001). By 7 days, the H2O2 production of cells adherent to ePTFE had decreased while that on cells adherent to SE was unchanged. These data indicate that the air pouch is a good alternative to other in vivo models for studies of the biological reactions induced by biomaterials. PMID- 8421001 TI - Extensive porosity at the cement-femoral prosthesis interface: a preliminary study. AB - This study is the first description of the extensive porosity which is preferentially located at the cement-prosthesis interface of cemented femoral components of total hip replacements. The observation is important because the interfacial porosity may decrease the strength of the cement-femoral prosthesis interface and jeopardize the mechanical integrity of the cement mantle. We examined the cement-metal interfaces from a multiplicity of in vivo and in vitro specimens using both optical and scanning electron microscopy. These samples included several stem designs, implants made from either Co-Cr or Ti alloy, implants made with a variety of surface finishes and both centrifuged and uncentrifuged cement. All in vivo and in vitro samples had marked porosity in the cement focally concentrated at the cement-metal interface. The amount of porosity at the interface greatly exceeded the amount of general porosity found throughout the bulk cement. Centrifuging did not affect the interfacial porosity, and neither did alloy nor surface finish. The presence of these pores may be explained by the rheological characteristics of the cement. PMID- 8421002 TI - A new type of biomaterial for artificial skin: dehydrothermally cross-linked composites of fibrillar and denatured collagens. AB - A new type of biomaterial for artificial skin was developed as a form of sponge by combining fibrillar collagen (F-collagen) with gelatin. The sponge was physically and metabolically stabilized by introducing dehydrothermal cross links. To get the final product, various conditions in the preparation of sponges were evaluated by in vitro cellular responses and in vivo tissue reactions. Fibroblasts placed on a sponge of gelatin attached themselves to it, migrated well into the sponge, and remained inside it for at least 7 days. However, sponges of gelatin showed structural instability for hydrolytic degradation by the cells. Most fibroblasts appeared not to penetrate into the interior of a sponge of F-collagen but to remain on its surface when fibroblasts were placed on the sponge, suggesting poor attraction of F-collagen toward cells. Implantation experiments of sponges of F-collagen revealed an intense infiltration of neutrophils into the sponge, indicating F-collagen as an inducer of the inflammatory reaction. These aggravating characters of F-collagen sponges were greatly improved by blending gelatin with F-collagen. The new type of collagen based biomaterials developed in the present study is expected to become a useful matrix substance for artificial skin. PMID- 8421003 TI - Thrombin and albumin adsorption to PVA and heparin-PVA hydrogels. 2: Competition and displacement. AB - Thrombin adsorption to polyvinyl alcohol (PVA) was different from its adsorption to polyethylene (PE)--not so much in amount, but in its affinity. Thrombin was more easily displaced from polyethylene and its adsorption was more readily prevented by prior or simultaneous exposure to albumin. From PVA (or heparin PVA), only approximately 30% of the adsorbed protein could be removed by a series of eluents, including even harsh ones such as 2.5M NaOH and 6M guanidine; > 85% could be removed from PE. Thrombin adsorption to PVA was not affected by the presence of BSA in solution or at the surface, but was virtually prevented on PE by preexposure to or adsorption with BSA. Heparin-PVA was not much different than PVA in most of these experiments, but did exhibit a "Vroman effect". In the absence of fibrinogen or antithrombin III, there was a maximum in thrombin adsorption from plasma at a plasma concentration of 1%. The behavior on this surface was dependent on both exposure time and protein concentration. These studies highlight the complexity of the interaction between plasma proteins and polymer surfaces (particularly hydrogel surfaces) and the difficulty of obtaining a clear picture of what happens when a single protein interacts with a polymer in the presence of other proteins. PMID- 8421004 TI - Enzyme-biomaterial interactions: effect of biosystems on degradation of polyurethanes. AB - Enzyme-induced liberation of hard-segment-containing components from polyurethanes was evaluated using two 14C-labeled polyurethanes. A polyester urea urethane and polyether urea-urethane were synthesized from toluene-2,4 diisocyanate (TDI)/polycaprolactone diol (PCL) or TDI/polyethylene glycol (PEO) with 14C-labeled ethylene diamine. Both materials were characterized using electron spectroscopy for chemical analysis (ESCA), differential scanning calorimetry (DSC), size exclusion chromatography, and material chemistry by Fourier transform infrared (FTIR) spectroscopy. Biodegradation assays were carried out using cholesterol esterase (CE), collagenase (CO), cathepsin B (CB), and xanthine oxidase (XO) at the pH optimum conditions for each enzyme at 37 degrees C. Biodegradation was analyzed by monitoring the release of radiolabel, by weight change, and by surface analysis using scanning electron microscopy. The polyester urea-urethane was shown to be susceptible to enzymatic degradation above the effect of the buffer control solution by the CE but not by the other enzyme systems as monitored by radiolabel released. In the initial period of incubation, the rate of degradation was increased for all systems, including buffer controls; however, the rates dropped off rapidly by day 28. The change in weight data for the polyester urea-urethane and polyether urea-urethane showed no enzyme-dependent biodegradation above the buffer controls. However, in sodium acetate buffer at pH = 5, the polymers showed a significant weight loss relative to other buffers. In conclusion, this study showed that the biological component responsible for the onset of the biodegradation process is more likely the result of a multitude of biologically mediated compounds acting synergistically, with the process being enhanced by physical parameters such as material dissolution. In addition characterization of surface and bulk chemistry as well as material structure evaluation have been shown to be essential to interpret degradation data. PMID- 8421005 TI - Bone induction in monkeys by bone morphogenetic protein. A trans-filter technique. AB - We investigated bone induction by bone morphogenetic protein in primates, comparing it with that seen in rodents. Twelve Millipore diffusion chambers containing 5 mg of semipurified bone morphogenetic protein were implanted into the dorsal muscles of 12 young, adult crab-eating monkeys (Macaca fascicularis) and were retrieved six weeks later. In six of nine unbroken chambers, new bone with haematopoietic marrow had been induced on the host-side surface of the filter. The incidence of trans-filter bone induction in the monkeys was almost equal to that observed in mice, and the new bone yield was approximately half as much as in mice. Our results show that bone morphogenetic protein can induce new bone formation in primates within six weeks, and support the hope that it will be useful as a substitute for bone graft in man. PMID- 8421006 TI - Minimally invasive surgery for osteoid osteoma of the proximal femur. AB - Seven patients with osteoid osteoma of the proximal femur were treated by percutaneous excision of the nidus. The combination of preoperative localisation by tomography and intraoperative localisation by image intensifier resulted in a curative procedure with minimal bone resection in all cases, although a second operation was required in one patient. PMID- 8421007 TI - Reconstruction nailing for pathological subtrochanteric fractures with coexisting femoral shaft metastases. AB - We describe the use of intramedullary reconstruction nails in the treatment of 14 patients with pathological subtrochanteric fractures and coexisting metastases in the femoral shaft. After nailing, all patients were free from pain and regained mobility. They were followed up clinically and radiologically until death from the primary disease. There were no mechanical failures even when a less than ideal reduction had been achieved. PMID- 8421008 TI - Vascularised fibular grafts for reconstruction of the femur. AB - From 1979 to 1990 we treated 20 patients with large bone defects or established nonunion of the femur by vascularised fibular grafts. There were 18 men and two women with an average age at operation of 36.6 years (16 to 69). Ten patients had infected nonunion, three had post-traumatic nonunion or a bone defect without infection, four had a defect after tumour resection, and three had other lesions. The mean length of the fibular grafts was 18.1 cm. Postoperative circulatory disturbances needed revision surgery in five patients, including three with circulatory problems in the monitoring flap, but not at their anastomoses. The outcome was successful in 19 of the 20 patients with bone union at means of 6.1 months at the proximal site and 6.6 months at the distal site. Three patients had fractures of the fibular grafts but all these united in two to three months after cast immobilisation. PMID- 8421009 TI - Avoiding nerve damage during elbow arthroscopy. AB - To define the anatomical relationships of the nerves to the common arthroscopy portals at the elbow an arthroscope was introduced into 20 cadaver elbows and the positions of the nerves were then determined by dissection. In all cases the posterior interosseous nerve lay close to the radiohumeral joint and to the anterolateral portal. Pronation of the forearm displaced the nerve away from the arthroscope. The median nerve passed consistently within 14 mm of the arthroscope when it was introduced through the anteromedial portal. The branches supplying the superficial forearm flexor muscles were at risk. PMID- 8421010 TI - Vascularity of the humeral head after proximal humeral fractures. An anatomical cadaver study. AB - We studied the arterial anatomy and the effect of four-part fractures on the vascularity of the humeral head, using barium sulphate perfusion of 16 cadaver shoulders. The main arterial supply to the humeral head was via the ascending branch of the anterior humeral circumflex artery and its intraosseous continuation, the arcuate artery. There were significant intraosseous anastomoses between the arcuate artery and: 1) the posterior humeral circumflex artery through vessels entering the posteromedial aspect of the proximal humerus; 2) metaphyseal vessels; and 3) the vessels of the greater and lesser tuberosities. Simulated four-part fractures prevented the perfusion of the humeral head in most cases. If, however, the head fragment extends distally below the articular surface medially, some perfusion of the head persists by the posteromedial vessels. These vessels are important in the management of comminuted fractures of the proximal humerus. PMID- 8421011 TI - The relative strengths of the rotator cuff muscles. A cadaver study. AB - We studied five cadaver shoulders to determine the strength relationship of the four rotator cuff muscles. The mean fibre length and volume of each muscle were measured, from which the physiological cross-sectional area was calculated. This value was used to estimate the force which each muscle was capable of generating. The lever arm of each muscle about the humeral head was then measured and the moment exerted was calculated. The strength ratios between the muscles were more or less constant in the five specimens. Subscapularis was the most powerful muscle and contributed 53% of the cuff moment; supraspinatus contributed 14%, infraspinatus 22% and teres minor 10%. The force-generating capacity of the subscapularis was equal to that of the other three muscles combined. PMID- 8421012 TI - Impacted cancellous allografts and cement for revision total hip arthroplasty. AB - We report the results of using impacted cancellous allografts and cement for fixation of the femoral component when revision arthroplasty is required in the face of lost bone stock. In 56 hips reviewed after 18 to 49 months there were few complications and a majority of satisfactory results with evidence of incorporation of the graft. Further study and review are necessary, but the use of the method appears to be justified. PMID- 8421013 TI - Clinical and pathological aspects of solitary spinal neurofibroma. AB - Four cases are described of solitary spinal neurofibroma, a rare tumour of the spinal cord or nerve roots. Computerised tomography provided an accurate topographical definition of the tumour. Magnetic resonance imaging showed an increased T2-weighted signal and multiple areas of decreased T1- and T2-weighted signals centrally. The MR signals matched the histological examination which showed hyperplastic interfascicular connective tissue, pleomorphic cells, and tightly packed nerve fibres compressed by the surrounding loose connective tissue. Electron microscopy showed three types of cell: Schwann cells, fibroblast like cells, and mast cells. The histological findings suggests that solitary spinal neurofibroma is a distinct pathological entity which could be diagnosed preoperatively from the MR images. PMID- 8421014 TI - Giant-cell tumours of the spine. AB - Between 1955 and 1989 we treated 24 patients (17 women and seven men) with giant cell tumours of the spine at the Mayo Clinic. Their mean age was 30 years and the mean follow-up time was 12.4 years. Pain was the presenting symptom in all and half had a neurological deficit. The cervical, thoracic, and lumbar spines were equally involved. The tumours recurred in five of the 14 patients treated by one stage surgery and in five of the ten treated by two-stage surgery. Seven patients received adjuvant radiotherapy, one for the primary lesion and six for recurrent lesions. Surgical management was by curettage or en bloc excision depending on the location and the extent of the tumour. Because of the risk of sarcomatous transformation, radiation therapy should be reserved for patients with incomplete excision or for those with local recurrence. PMID- 8421015 TI - Callotasis in melorheostosis: a case report. PMID- 8421016 TI - Diastasis between the medial and the intermediate cuneiforms. PMID- 8421017 TI - Regional prophylactic antibiotic in knee arthroplasty. PMID- 8421018 TI - Avascular necrosis of the femoral head in HIV-infected patients. PMID- 8421019 TI - Particles in loose hips. PMID- 8421020 TI - Hip socket impingement. PMID- 8421021 TI - Footprints and arches. PMID- 8421022 TI - Hip fractures and QALYS. PMID- 8421023 TI - Standardisation of long radiographs. PMID- 8421024 TI - Locked nailing of humeral fractures. PMID- 8421025 TI - Gaucher's disease: a novel treatment and an important breakthrough. PMID- 8421026 TI - Sequential haematogenous infection of bilateral cementless total hip replacements. AB - We report a patient with bilateral uncemented total hip replacements in whom a deep infection on the second side may have been caused by a transient bacteraemia associated with a revision operation for deep infection on the first side. Both hips were successfully treated by one-stage exchange. PMID- 8421027 TI - A technique for removing an intrapelvic acetabular cup. AB - We describe a simple, retroperitoneal approach for the removal of acetabular components that have migrated into the pelvis. The dense fibrous tissue layer which surrounds the implant protects the iliac vessels during removal of the cup by this method. PMID- 8421028 TI - Exchange arthroplasty for infected knee replacements. A new two-stage method. AB - We report a series of 17 exchange arthroplasties for infected knee prostheses, ten one-stage and seven two-stage procedures. The method proved successful in controlling infection and restoring function. In two-stage exchanges the interval between the stages was managed by using a prosthesis as a spacer, and acrylic cement beads containing the appropriate antibiotic to provide high local concentrations. Three one-stage procedures had recurrence of infection, but were successfully treated by further exchange operations. All patients had satisfactory function and there have been no serious complications. We recommend this modified two-stage technique for the management of infected knee arthroplasties. PMID- 8421029 TI - Timing of antibiotic administration in knee replacement under tourniquet. AB - Cephamandole levels in serum and drain fluid were measured in 32 knee replacement operations to determine the benefit of an intravenous dose of antibiotic at the time of tourniquet deflation. Concentrations of cephamandole in drain fluid were directly proportional to the serum concentration at the time of tourniquet release. A 'tourniquet-release' dose of antibiotic increased drain fluid concentration threefold. PMID- 8421030 TI - Results of partial meniscectomy related to the state of the anterior cruciate ligament. Review at 20 to 35 years. AB - We reviewed 195 knees in 167 patients at least 20 years after a rim-preserving meniscectomy. They were considered in two groups: 102 knees had had an intact anterior cruciate ligament (ACL), and 93 had had an unrepaired rupture. More patients with a ruptured ACL had downgraded their sport activity by five years after meniscectomy. The incidence of radiographic osteoarthritis was about 65% at 27 years in patients with a ruptured ligament, and 86% in those followed up for over 30 years. In the ligament-deficient group 10% had had operations for osteoarthritis, and another 28% had had other operations, mainly further meniscectomies. Only 6% of those with an intact ligament had needed a second operation after meniscectomy and at long-term follow-up 92% of them were satisfied or very satisfied. Only 74% of the ligament-deficient patients were satisfied with their result. The long-term outcome after rim-preserving meniscectomy depends mainly upon the state of the anterior cruciate ligament. PMID- 8421031 TI - Ionising radiation and the orthopaedic patient. PMID- 8421032 TI - Can muscle co-contraction protect knee ligaments after injury or repair? AB - A computer-based model of the knee was used to study forces in the cruciate ligaments induced by co-contraction of the extensor and flexor muscles, in the absence of external loads. Ligament forces are required whenever the components of the muscle forces parallel to the tibial plateau do not balance. When the extending effect of quadriceps exactly balances the flexing effect of hamstrings, the horizontal components of the two muscle forces also balance only at the critical flexion angle of 22 degrees. The calculations show that co-contraction of the quadriceps and hamstring muscles loads the anterior cruciate ligament from full extension to 22 degrees of flexion and loads the posterior cruciate at higher flexion angles. In these two regions of flexion, the forward pull of the patellar tendon on the tibia is, respectively, greater than or less than the backward pull of hamstrings. Simultaneous quadriceps and gastrocnemius contraction loads the anterior cruciate over the entire flexion range. Simultaneous contraction of all three muscle groups can unload the cruciate ligaments entirely at flexion angles above 22 degrees. These results may help the design of rational regimes of rehabilitation after ligament injury or repair. PMID- 8421033 TI - Can MRI of the knee affect arthroscopic practice? A prospective study of 58 patients. AB - We made a prospective study of 58 patients with suspected internal derangement of the knee. They were examined by magnetic resonance imaging using 3-D gradient echo intermediate-weighted studies before having an arthroscopy. The preoperative clinical assessment was found to have a diagnostic sensitivity of 77% and a specificity of 43%, compared with 100% and 63% respectively for magnetic resonance imaging. Comparison of magnetic resonance imaging and arthroscopy confirmed the accuracy of magnetic resonance imaging in the diagnosis of internal derangement but the results for articular cartilage lesions were much less good, with a sensitivity of only 18% but a specificity of 100%. Acceptance of the magnetic resonance imaging findings could have resulted in a 29% reduction in the number of arthroscopies without missing any significant meniscal lesion. PMID- 8421034 TI - Poor results of Darrach's procedure after wrist injuries. AB - Darrach's procedure was performed for post-traumatic symptoms in the inferior radio-ulnar joint in 36 patients, who were reviewed after a mean follow-up of 6 years. Only 18 of the patients had a satisfactory clinical result. Poor outcome was associated with osteoarthritis of the wrist, the occurrence of algodystrophy and a short ulnar remnant. PMID- 8421035 TI - The distal radio-ulnar joint in Colles' fractures. AB - The effect of involvement of the distal radio-ulnar joint on subsequent function was studied in 170 patients with Colles' fractures, reviewed for one year after cast removal. Patients with involvement of this joint had significantly weaker grips and a significantly greater incidence of pain and tenderness over the joint at all stages of follow-up. They also had a poorer range of supination at six months and at one year. The presence or absence of an ulnar styloid fracture was not related to the functional results. PMID- 8421036 TI - Uncemented porous-coated anatomic total hip replacement. Results at six years in a consecutive series. AB - We studied 108 patients (116 hips) who were followed for a minimum of six years (73 to 89 months) after primary total hip arthroplasty using an uncemented porous coated anatomic hip. The average age of the patients at operation was 48.4 years, and the diagnosis was avascular necrosis of the femoral head in 46 hips, neglected femoral neck fracture in 27, osteoarthritis secondary to childhood pyogenic arthritis in 24 and to childhood tuberculous arthritis in five, and miscellaneous in 14. The average preoperative Harris hip score was 55, which improved to 91 at latest follow-up. All patients with loose femoral components or disabling thigh pain had received prostheses which were undersized in the coronal or the sagittal plane, or in both. No patient with a satisfactory fit in both coronal and sagittal planes had loosening of the femoral component or disabling thigh pain. Three acetabular components showed aseptic loosening and 20 showed excessive wear (5 to 11 mm) of the polyethylene liner. Excessive wear was related to young age, but not to body-weight, gender, primary diagnosis, hip score, or range of hip movement. There was a high incidence of osteolysis (38 of 116 hips, 33%). We recommend careful preoperative planning, with the use of a contemporary cemented technique when a satisfactory fit cannot be obtained. The high incidence of excessive wear and of osteolysis needs further investigation. PMID- 8421037 TI - Radiography and scintigraphy of suspected scaphoid fracture. A long-term study in 160 patients. AB - Radiographs of the scaphoid after injury are difficult to interpret, and bone scintigraphy is widely used to increase the accuracy of diagnosis, though many fractures suspected on scintigraphy cannot be confirmed radiologically. We have reviewed the clinical consequences, after one year, of managing suspected scaphoid fractures according to the bone-scan results. We studied 160 patients, 35 of whom had initially positive radiographs and were treated in a cast for 12 weeks. The other 125 had bone scintigraphy and were managed according to the result. After a minimum of one year 119 patients were reviewed. Scintigraphically suspected scaphoid fracture could not be confirmed radiologically in 25%. There were no cases of nonunion. The long period of immobilisation in patients with positive radiographs or positive bone scans did not influence the frequency or severity of late symptoms compared with those with a normal bone scan. PMID- 8421038 TI - Ultrasound or image intensifier for closed femoral nailing. AB - We describe a method of closed, unlocked nailing for femoral fractures using ultrasound instead of an image intensifier. Radiography was used only to confirm that the guide wire had been passed into the intramedullary canal of both fragments. The method succeeded in 26 of 30 cases. The failures all occurred in fractures which could not be reduced within 20 minutes. The operating time in those nailed successfully with ultrasound control was not different from the time for 30 control cases using conventional methods with an image intensifier. PMID- 8421039 TI - A new technique of talectomy for severe fracture-dislocation of the talus. AB - We report a new technique of talectomy for patients with Hawkins group III fracture-dislocation of the talus. Talectomy is performed through a medial incision, the foot is displaced anteriorly, and the fractured or osteotomised medial malleolus is moved laterally and fixed to the tibia with a malleolar screw. Full weight-bearing is allowed after six weeks. In four patients at 36 to 57 months after operation the results were excellent in three and good in one, with no pain or early evidence of degenerative arthritis in the remaining joints of the foot. PMID- 8421040 TI - Failures of screening and management of congenital dislocation of the hip. AB - We report the screening of 67,093 infants for congenital dislocation of the hip from 1980 to 1989 and compare the results with those during the preceding two decades. More dislocations have been missed at neonatal examination during the last decade (0.13% of live births). Operative treatment was needed in 54 children (0.08% of live births) some of whom had been diagnosed at birth. We discuss the reasons for the failure of neonatal screening. PMID- 8421041 TI - Joint laxity and hip rotation in normal children and in those with congenital dislocation of the hip. AB - We measured the range of rotation in both hips of 397 normal children and in the unaffected hip of 135 children with unilateral congenital dislocation of the hip. Both groups were assessed for generalised joint laxity. Joint laxity was more common in normal children with an internally centred arc of hip rotation than in normal children with a neutral or an externally rotated arc. The children with congenitally dislocated hips had significantly more joint laxity than did the control group and significantly more of them had an internally centred arc of hip rotation. We suggest that the lax joint capsule fails to mould away the neonatal anteversion of the femoral neck during the first few months of life. PMID- 8421042 TI - Ultrasound in diagnosis and management of acute haematogenous osteomyelitis in children. AB - We reviewed the ultrasound findings in 59 children suspected of having bone infection. Twenty-nine were eventually proved to have acute haematogenous osteomyelitis and 26 of these showed characteristic ultrasound findings. Such changes were rare in 30 patients with other clinically similar conditions. Ultrasound examination was also able to detect the presence of subperiosteal pus and thus indicate the need for surgical treatment. PMID- 8421043 TI - Paralytic drop foot and gluteal fibrosis after intramuscular injections. AB - Eight children with paralytic drop foot after intramuscular injections later developed gluteal fibrosis. Sciatic palsy, presenting as equinovarus or equinus deformity, was diagnosed on average 3.8 months after the intragluteal injections, but gluteal fibrosis was not diagnosed until 5.1 years after the injections. In three patients the equinovarus recurred after surgical correction due to persistent muscle imbalance and the effect of the external rotation contracture of the hip. PMID- 8421044 TI - A chart of anthropometric values. AB - A chart is presented to assist with the assessment and treatment of patients with growth-related deformities. It is based on anthropometric values from five published sources and relates sitting height to stature, limb length, the radiographic lengths of the leg bones and the lengths of the feet and hands. It has proved useful in the prediction of leg-length discrepancies, in the diagnosis of cases of short stature, and in the assessment of spinal shortening from scoliosis. PMID- 8421045 TI - Laboratory comparison of the cannulated Herbert bone screw with ASIF cancellous lag screws. AB - The compression produced by and the resistance to pullout of the 6.5 mm cannulated Herbert screw were compared with those of ASIF headed screws. The latter were tested with and without washers and in the following sizes: 4.5 mm cortical, 6.5 mm cancellous with a 16 mm threaded segment, and 6.5 mm cancellous with a 32 mm threaded segment. Polyurethane foam was used as a substitute for cancellous bone and ASIF artificial bone for corticocancellous bone. The compression produced by a cancellous lag screw with a washer was significantly greater than that produced by a Herbert screw of equivalent size (p < 0.05). When the screws were tested using the corticocancellous composite the ASIF cancellous screw without a washer produced significantly greater compression (p < 0.05); when used with a washer the difference was highly significant (p < 0.001). The dual pitch Herbert screw is not appropriate for the management of fractures in which compression is of greater importance than the need to avoid prominence of the screw head. PMID- 8421046 TI - Prenatal development of the iliolumbar ligament. AB - Serial sections, in the frontal plane, of 12 human fetuses showed that the iliolumbar ligament was always present at the gestational age of 11 to 15 weeks; in younger specimens, it could not be identified. The ligament develops during the prenatal period and is not formed by metaplasia of the quadratus lumborum muscle during the second decade of life. PMID- 8421047 TI - New bone formation during leg lengthening. Evaluated by dual energy X-ray absorptiometry. AB - We measured the extent and rate of new bone formation over an 18-month period before, during and after the lengthening of ten leg segments in six patients aged between 8 and 18 years, using dual-energy X-ray absorptiometry (DEXA). New bone formation could be identified within one week of the start of distraction. As lengthening proceeded, the bone density of the gap fell, reaching minimum values at the time of maximal distraction. Consolidation of the regenerating bone was started 1 to 2 weeks later in the tibia, and 2.5 to 3.0 weeks later in the femur. The rate of mineral accretion in new bone was significantly greater in the tibia than in the femur (16 +/- 1.86%/month, and 11 +/- 1.1%/month respectively; mean +/- SEM). There was significant osteoporosis distal to the osteotomy, more in the tibia than in the femur, particularly on the side of the fixator. The bone mineral density of the distal segment remained low at the time of fixator removal (44.2 +/- 5.58% and 61.0 +/- 4.2% of the control values at the tibia and femur respectively) and was only partially reversed by subsequent weight-bearing. We conclude that dual-energy X-ray absorptiometry provides an objective and quantitative assessment of new bone formation during leg lengthening. The technique also allows the measurement of the distraction gap and the assessment of leg alignment from the high-resolution images. Its use may decrease the requirements for conventional radiography. PMID- 8421048 TI - Histone H4 acetylation and transcription in amphibian chromatin. AB - Lampbrush chromosomes from oocytes of the amphibian Triturus cristatus have been used to examine the role of histone acetylation in transcription by indirect immunofluorescence with antisera to H4 acetylated at specific lysine residues. Electrophoresis on acid-urea-Triton gels and Western blotting have confirmed the specificity of these antisera and defined the order in which particular lysine residues are acetylated in amphibian cells. As in mammals, lysine 16 is acetylated first, followed by 8 and/or 12 and then 5. With lampbrush chromosomes from immature (previtellogenic) oocytes, antisera to H4 acetylated at lysines 8, 12, and 16 labeled fluorescent foci at the bases of transcription loops. Antisera to H4 acetylated at lysine 5 labeled weakly (i.e., the tri- and tetraacetylated isoforms must be rare). Loops showed weak labeling of the chromatin axis but intense fluorescence at particular points, which probably represent incompletely decondensed chromatin. The RNP matrix of loops, including the RNP-rich sphere bodies and the dense matrix of "marker" loops, was not labeled. Treatment of immature oocytes with butyrate for 12 h to inhibit histone deacetylation did not affect immunolabeling, suggesting that turnover of H4 acetates is slow. In contrast, in chromosomes from mature oocytes, in which loops have retracted and transcription is low, butyrate caused an increase in labeling with all antisera, followed by the appearance of vestigial loops, weakly labeled, but with regions of intense fluorescence. These loops contain RNP and are presumably transcriptionally active. We conclude that H4 acetates turn over more rapidly in mature than immature oocytes and that histone hyperacetylation precedes, and possibly induces, loop formation and transcriptional activation. PMID- 8421049 TI - Overlapping domains of the heterochromatin-associated protein HP1 mediate nuclear localization and heterochromatin binding. AB - The Drosophila protein HP1 is a 206 amino acid heterochromatin-associated nonhistone chromosomal protein. Based on the characterization of HP1 to date, there are three properties intrinsic to HP1: nuclear localization, heterochromatin binding, and gene silencing. In this work, we have concentrated on the identification of domains responsible for the nuclear localization and heterochromatin binding properties of HP1. We have expressed a series of beta galactosidase/HP1 fusion proteins in Drosophila embryos and polytene tissue and have used beta-galactosidase enzymatic activity to identify the subcellular localization of each fusion protein. We have identified two functional domains in HP1: a nuclear localization domain of amino acids 152-206 and a heterochromatin binding domain of amino acids 95-206. Both of these functional domains overlap an evolutionarily conserved COOH-terminal region. PMID- 8421050 TI - Structure of the colcemid-treated PtK1 kinetochore outer plate as determined by high voltage electron microscopic tomography. AB - High voltage electron microscopic tomography was used to determine the organization of the kinetochore plate and its attachment to the underlying chromosome. Six reconstructions were computed from thick sections of Colcemid treated PtK1 cells and analyzed by a number of computer graphics methods including extensive thin slicing, three-dimensional masking, and volume rendering. When viewed en-face the kinetochore plate appeared to be constructed from a scaffold of numerous 10-20-nm thick fibers or rods. Although the fibers exhibited regions of parallel alignment and hints of a lattice, they were highly variable in length, orientation and spacing. When viewed in stereo, groups of these fibers were often seen oriented in different directions at different depths to give an overall matted appearance to the structure. When viewed "on edge," the plate was 35-40 nm thick, and in thin slices many regions were tripartite with electron-opaque domains, separated by a more translucent middle layer, forming the inner and outer plate boundaries. These domains were joined at irregular intervals. In some slices, each domain appeared as a linear array of 10-20-nm dots or rods embedded in a less electron-opaque matrix, and adjacent dots within or between domains often appeared fused to form larger blocks. The plate was connected to the underlying chromosome by less densely arrayed 10-20-nm thick fibers that contacted the chromosome-facing (i.e., inner) surface of the plate in numerous patches. These patches tended to be arrayed in parallel rows perpendicular to the long axis of the chromosome. In contrast to connecting fibers, corona fibers were more uniformly distributed over the cytoplasmic-facing (i.e., outer) surface of the plate. When large portions of the reconstructions were viewed, either en-face or in successive slices parallel to the long axis of the chromosome, the edges of the plate appeared splayed into multiple "fingers" that partly encircled the primary constriction. Together these observations reveal that regions of the kinetochore outer plate contain separate structural domains, which we hypothesize to serve separate functional roles. Our three dimensional images of the kinetochore are largely consistent with the hypothesis that the outer plate is composed of multiple identical subunits (Zinkowski, R. P., J. Meyne, and B. R. Brinkley. 1991. J. Cell Biol. 113:1091-1110). PMID- 8421051 TI - Ran/TC4: a small nuclear GTP-binding protein that regulates DNA synthesis. AB - Ran/TC4, first identified as a well-conserved gene distantly related to H-RAS, encodes a protein which has recently been shown in yeast and mammalian systems to interact with RCC1, a protein whose function is required for the normal coupling of the completion of DNA synthesis and the initiation of mitosis. Here, we present data indicating that the nuclear localization of Ran/TC4 requires the presence of RCC1. Transient expression of a Ran/TC4 protein with mutations expected to perturb GTP hydrolysis disrupts host cell DNA synthesis. These results suggest that Ran/TC4 and RCC1 are components of a GTPase switch that monitors the progress of DNA synthesis and couples the completion of DNA synthesis to the onset of mitosis. PMID- 8421052 TI - An integral membrane glycoprotein associated with an endocytic compartment of Trypanosoma vivax: identification and partial characterization. AB - A 65-kD glycoprotein (gp65) of Trypanosoma (Duttonella) vivax was identified using a murine monoclonal antibody (mAb 4E1) that had been raised against formalin-fixed, in vitro-propagated, uncoated forms. Intracellular localization studies utilizing the mAb in immunofluorescence on fixed, permeabilized T. vivax bloodstream forms and immunoelectron microscopy on thin sections of Lowicryl K4M embedded cells revealed labeling of vesicles and tubules in the posterior portion of the parasite. Some mAb-labeled vesicles contained endocytosed 10 nm BSA-gold after incubation of the parasites with the marker for 5-30 min at 37 degrees C, and the greatest degree of colocalization was observed after 5 min. Double labeling experiments using the mAb and a polyclonal anti-variant surface glycoprotein (VSG) antibody to simultaneously localize both gp65 and VSG demonstrated that there was little overlap in the distribution of these antigens. Thus, gp65 is associated with tubules and vesicles that are involved in endocytosis but which appear to be distinct from VSG processing pathways within the cell. Using the mAb for immunoblot analyses, gp65 was shown to be enriched in a fraction of solubilized membrane proteins eluted from either immobilized Con A or Ricinus communis agglutinin and was found to possess carbohydrate linkages cleaved by both endoglycosidase H and O-glycosidase, suggesting the presence of N and O-linked glycans. Protease protection and crosslinking experiments suggest that gp65 is a transmembrane protein with trypsin cleavage and NH2-crosslinking sites on the lumenal face of the vesicles. PMID- 8421053 TI - Cloning, functional analysis and cell localization of a kidney proximal tubule water transporter homologous to CHIP28. AB - The localization and transporting properties of a kidney protein homologous to human erythrocyte protein CHIP28 was evaluated. The cDNA encoding rat kidney protein CHIP28k was isolated from a rat renal cortex cDNA library. A 2.8-kb cDNA was identified which contained an 807 bp open reading frame encoding a 28.8 kD protein with 94% amino acid identity to CHIP28. in vitro translation of CHIP28k cDNA in rabbit reticulocyte lysate generated a 28-kD protein; addition of ER derived microsomes gave a 32-kD transmembrane glycoprotein. Translation of truncated RNA demonstrated glycosylation of residue Asn42 which is predicted to lie between the first and second transmembrane domains. Expression of in vitro transcribed mRNA encoding CHIP28k in Xenopus oocytes increased oocyte osmotic water permeability (Pf) from (4 +/- 1) x 10(-4) to (33 +/- 4) x 10(-4) cm/s at 10 degrees C; the increase in oocyte Pf was weakly temperature dependent and inhibited by HgCl2. Two-electrode voltage clamp measurements indicated that CHIP28k was not permeable to ions. Oocyte Pf also increased with expression of total mRNA from kidney cortex and papilla; the increase in Pf with mRNA from cortex, but not kidney papilla, was blocked by coinjection with excess antisense CHIP28k cRNA. In situ hybridization of a 150 base cRNA antisense probe to tissue sections from rat kidney showed selective CHIP28k localization to epithelial cells in proximal tubule and thin descending limb of Henle. Pf in purified apical membrane vesicles from rat and human proximal tubule, and in proteoliposomes reconstituted with purified protein, was very high and inhibited by HgCl2; stripping of apical vesicles with N-lauroylsarcosine enriched a 28-kD protein by 25-fold and yielded a vesicle population with high water, but low urea and proton permeabilities. CHIP28k identity was confirmed by NH2-terminus sequence analysis. These results indicate that CHIP28k is a major and highly selective water transporting protein in the kidney proximal tubule and thin descending limb of Henle, but not collecting duct. PMID- 8421054 TI - A family of cation ATPase-like molecules from Plasmodium falciparum. AB - We report the nucleotide and derived amino acid sequence of the ATPase 1 gene from Plasmodium falciparum. The amino acid sequence shares homology with the family of "P"-type cation translocating ATPases in conserved regions important for nucleotide binding, conformational change, or phosphorylation. The gene, which is present on chromosome 5, has a product longer than any other reported for a P-type ATPase. Interstrain analysis from 12 parasite isolates by the polymerase chain reaction reveals that a 330-bp nucleotide sequence encoding three cytoplasmic regions conserved in cation ATPases (regions a-c) is of constant length. By contrast, another 360-bp sequence which is one of four regions we refer to as "inserts" contains arrays of tandem repeats which show length variation between different parasite isolates. Polymorphism results from differences in the number and types of repeat motif contained in this insert. Inserts are divergent in sequence from other P-type ATPases and share features in common with many malarial antigens. Studies using RNA from the erythrocytic stages of the malarial life cycle suggest that ATPase 1 (including the sequence which encodes tandem repeats) is expressed at the large ring stage of development. Immunolocalization has identified ATPase 1 to be in the region of the parasite plasma membrane and pigment body. These findings suggest a possible model for the genesis of malarial antigens. PMID- 8421055 TI - Tropomodulin is associated with the free (pointed) ends of the thin filaments in rat skeletal muscle. AB - The length and spatial organization of thin filaments in skeletal muscle sarcomeres are precisely maintained and are essential for efficient muscle contraction. While the major structural components of skeletal muscle sarcomeres have been well characterized, the mechanisms that regulate thin filament length and spatial organization are not well understood. Tropomodulin is a new, 40.6-kD tropomyosin-binding protein from the human erythrocyte membrane skeleton that binds to one end of erythrocyte tropomyosin and blocks head-to-tail association of tropomyosin molecules along actin filaments. Here we show that rat psoas skeletal muscle contains tropomodulin based on immunoreactivity, identical apparent mobility on SDS gels, and ability to bind muscle tropomyosin. Results from immunofluorescence labeling of isolated myofibrils at resting and stretched lengths using anti-erythrocyte tropomodulin antibodies indicate that tropomodulin is localized at or near the free (pointed) ends of the thin filaments; this localization is not dependent on the presence of myosin thick filaments. Immunoblotting of supernatants and pellets obtained after extraction of myosin from myofibrils also indicates that tropomodulin remains associated with the thin filaments. 1.2-1.6 copies of muscle tropomodulin are present per thin filament in myofibrils, supporting the possibility that one or two tropomodulin molecules may be associated with the two terminal tropomyosin molecules at the pointed end of each thin filament. Although a number of proteins are associated with the barbed ends of the thin filaments at the Z disc, tropomodulin is the first protein to be specifically located at or near the pointed ends of the thin filaments. We propose that tropomodulin may cap the tropomyosin polymers at the pointed end of the thin filament and play a role in regulating thin filament length. PMID- 8421056 TI - Cofilin is an essential component of the yeast cortical cytoskeleton. AB - We have biochemically identified the Saccharomyces cerevisiae homologue of the mammalian actin binding protein cofilin. Cofilin and related proteins isolated from diverse organisms are low molecular weight proteins (15-20 kD) that possess several activities in vitro. All bind to monomeric actin and sever filaments, and some can stably associate with filaments. In this study, we demonstrate using viscosity, sedimentation, and actin assembly rate assays that yeast cofilin (16 kD) possesses all of these properties. Cloning and sequencing of the S. cerevisiae cofilin gene (COF1) revealed that yeast cofilin is 41% identical in amino acid sequence to mammalian cofilin and, surprisingly, has homology to a protein outside the family of cofilin-like proteins. The NH2-terminal 16kD of Abp1p, a 65-kD yeast protein identified by its ability to bind to actin filaments, is 23% identical to yeast cofilin. Immunofluorescence experiments showed that, like Abp1p, cofilin is associated with the membrane actin cytoskeleton. A complete disruption of the COF1 gene was created in diploid cells. Sporulation and tetrad analysis revealed that yeast cofilin has an essential function in vivo. Although Abp1p shares sequence similarity with cofilin and has the same distribution as cofilin in the cell, multiple copies of the ABP1 gene cannot compensate for the loss of cofilin. Thus, cofilin and Abp1p are structurally related but functionally distinct components of the yeast membrane cytoskeleton. PMID- 8421057 TI - Concentration of an integral membrane protein, CD43 (leukosialin, sialophorin), in the cleavage furrow through the interaction of its cytoplasmic domain with actin-based cytoskeletons. AB - In leukocytes such as thymocytes and basophilic leukemia cells, a glycosilated integral membrane protein called CD43 (leukosialin or sialophorin), which is defective in patients with Wiskott-Aldrich syndrome, was highly concentrated in the cleavage furrow during cytokinesis. Not only at the mitotic phase but also at interphase, CD43 was precisely colocalized with ezrin-radixin-moesin family members. (ERM), which were previously reported to play an important role in the plasma membrane-actin filament association in general. At the electron microscopic level, throughout the cell cycle, both CD43 and ERM were tightly associated with microvilli, providing membrane attachment sites for actin filaments. We constructed a cDNA encoding a chimeric molecule consisting of the extracellular domain of mouse E-cadherin and the transmembrane/cytoplasmic domain of rat CD43, and introduced it into mouse L fibroblasts lacking both endogenous CD43 and E-cadherin. In dividing transfectants, the chimeric molecules were concentrated in the cleavage furrow together with ERM, and both proteins were precisely colocalized throughout the cell cycle. Furthermore, using this transfection system, we narrowed down the domain responsible for the CD43 concentration in the cleavage furrow. Based on these findings, we conclude that CD43 is concentrated in the cleavage furrow through the direct or indirect interaction of its cytoplasmic domain with ERM and actin filaments. PMID- 8421058 TI - Dynamics of microtubules bundled by microtubule associated protein 2C (MAP2C). AB - MAP2C is a microtubule-associated protein abundant in immature nerve cells. We isolated a cDNA clone encoding whole mouse MAP2C of 467 amino acid residues. In fibroblasts transiently transfected with cDNA of MAP2C, interphase microtubule networks were reorganized into microtubule bundles. To reveal the dynamic properties of microtubule bundles, we analyzed the incorporation sites of exogenously introduced tubulin by microinjection of biotin-labeled tubulin and the turnover rate of microtubule bundles by photoactivation of caged fluorescein labeled tubulin. The injected biotin-labeled tubulin was rapidly incorporated into distal ends of preexisting microtubule bundles, suggesting a concentration of the available ends of microtubules at this region. Although homogenous staining of microtubule bundles with antibiotin antibody was observed 2 h after injection, the photoactivation study indicated that turnover of microtubule bundles was extremely suppressed and < 10% of tubulin molecules would be exchanged within 1 h. Multiple photoactivation experiments provided evidence that neither catastrophic disassembly at the distal ends of bundles nor concerted disassembly due to treadmilling at the proximal ends could explain the observed rapid incorporation of exogenously introduced tubulin molecules. We conclude that microtubules bundled by MAP2C molecules are very stable while the abrupt increase of free tubulin molecules by microinjection results in rapid assembly from the distal ends within the bundles as well as free nucleation of small microtubules which are progressively associated laterally with preexisting microtubule bundles. This is the first detailed study of the function of MAPs on the dynamics of microtubules in vivo. PMID- 8421059 TI - Monoclonal antibody 7H6 reacts with a novel tight junction-associated protein distinct from ZO-1, cingulin and ZO-2. AB - The tight junction is an essential element of the intercellular junctional complex; yet its protein composition is not fully understood. At present, only three proteins, ZO-1 (Stevenson, B. R., J. D. Siliciano, M. S. Mooseker, and D. A. Goodenough. 1986. J. Cell Biol. 103:755-766), cingulin (Citi, S., H. Sabanay, R. Jakes, B. Geiger, and J. Kendrick-Jones. 1988. Nature (Lond.). 333:272-275) and ZO-2 (Gumbiner, B., T. Lowenkopf, and D. Apatira. 1991. Proc. Natl. Acad. Sci. USA. 88:3460-3464) are known to be associated with the tight junction. We have generated a monoclonal antibody (7H6) against a bile canaliculus-rich membrane fraction prepared from rat liver. This 7H6 antigen was preferentially localized by immunofluorescence at the junctional complex regions of hepatocytes and other epithelia, and 7H6-affiliated gold particles were shown electron microscopically to localize at the periphery of tight junctions. Immunoblot analysis of a bile canaliculus-rich fraction of rat liver using 7H6, anti-ZO-1 antibody (R26.4C), and anti-cingulin antibody revealed that 7H6 reacted selectively with a 155-kD protein, whereas R26.4C reacted only with a 225-kD protein. Anti-cingulin antibody reacted solely with 140 and 108-kD proteins, indicating that the protein recognized by 7H6 is immunologically different from ZO-1 and cingulin. Immunoprecipitation of detergent extracts obtained from metabolically labeled MDCK cells with R26.4C coprecipitated a 160-kD protein, which corresponds to ZO-2, with ZO-1. However, 7H6 did not react with the 160-kD protein. These results strongly suggest that the 7H6 antibody recognizes a novel tight junction-associated protein different from ZO-1, cingulin and ZO-2. PMID- 8421060 TI - Antennapedia homeobox peptide enhances growth and branching of embryonic chicken motoneurons in vitro. AB - Spinal motoneuron development is regulated by a variety of intrinsic and extrinsic factors. Among these, a possible role for homeoproteins is suggested by their expression in the motoneuron at relatively late stages. To investigate their possible involvement in motoneuron growth, we adapted a novel technique recently developed in this laboratory, based on the ability of the 60 amino acid long homeobox of Antennapedia (pAntp) to translocate through the neuronal membrane and to accumulate in the nucleus (Joliot, A. H., C. Pernelle, H. Deagostini-Bazin, and A. Prochiantz. 1991. Proc. Natl. Acad. Sci. USA. 88:1864 1868; Joliot, A. H., A. Triller, M. Volovitch, C. Pernelle, and A. Prochiantz. 1991. New Biol. 3:1121-1134). Motoneurons from E5 chicken spinal cord were incubated with pAntp, purified by panning on SC1 antibody and plated on polyornithine/laminin substrata without further addition of pAntp. After 24 h, neurite outgrowth was already extensive in controls. In cultures of motoneurons that had been preincubated with 10(-7) M pAntp, neurite length was doubled; a similar effect was obtained using postnatal muscle extracts. Morphological analysis using a neurofilament marker specific for axons indicated that the homeobox peptide enhances primarily axonal elongation and branching. To test the hypothesis that the biological activity of pAntp involves its specific attachment to cognate homeobox binding sites present in the genome, we generated a mutant of pAntp called pAntp40P2, that was still able to translocate through the motoneuron membrane and to reach the nucleus, but had lost the specific DNA-binding properties of the wild-type peptide. Preincubation of pAntp40P2 with purified motoneurons failed to increase neurite outgrowth. This finding raises the possibility that motoneuron growth is controlled by homeobox proteins. PMID- 8421061 TI - Biosynthesis and in vivo localization of the decapentaplegic-Vg-related protein, DVR-6 (bone morphogenetic protein-6). AB - DVR-6 (BMP-6 or Vgr-1) is a member of the TGF-beta superfamily of polypeptide signaling molecules. In situ hybridization studies have previously shown that DVR 6 RNA is expressed in a variety of cell types in the mouse embryo, but no information has been available on protein localization and biosynthesis. We have produced a polyclonal antibody to the proregion of DVR-6 and used it to localize the protein in whole mount and sectioned embryonic, newborn, and adult mouse tissues. DVR-6 protein is expressed in the mouse nervous system beginning at 9.5 days postcoitum (d.p.c.) and continues through adulthood. A variety of epithelial tissues also produce DVR-6 protein, including the suprabasal layer of the skin, bronchiolar epithelium, and the cornea. Additionally, a stably transfected cell line, BMGE+H/D6c4, is used to study the biosynthesis of DVR-6 protein and evidence is presented for translational regulation of DVR-6 expression. PMID- 8421062 TI - Phosphorylation-site mutagenesis of the growth-associated protein GAP-43 modulates its effects on cell spreading and morphology. AB - The 43-kD growth-associated protein (GAP-43) is a major protein kinase C (PKC) substrate of axonal growth cones, developing nerve terminals, regenerating axons, and adult central nervous system areas associated with plasticity. It is a cytosolic protein associated with the cortical cytoskeleton and the plasmalemma. Membrane association of GAP-43 is mediated by palmitoylation at Cys3Cys4. In vitro and in vivo, phosphorylation by PKC exclusively involves Ser41 of mammalian GAP-43 (corresponding to Ser42 in the chick protein). To identify aspects of GAP 43 function, we analyzed the actions of wild-type, membrane-association, and phosphorylation-site mutants of GAP-43 in nonneuronal cell lines. The GAP-43 constructs were introduced in L6 and COS-7 cells by transient transfection. Like the endogenous protein in neurons and their growth cones, GAP-43 in nonneuronal cells associated with the cell periphery. GAP-43 accumulated in the pseudopods of spreading cells and appeared to interact with cortical actin-containing filaments. Spreading L6 cells expressing high levels of recombinant protein displayed a characteristic F-actin labeling pattern consisting of prominent radial arrays of peripheral actin filaments. GAP-43 had dramatic effects on local surface morphology. Characteristic features of GAP-43-expressing cells were irregular cell outlines with prominent and numerous filopodia. The effects of GAP 43 on cell morphology required association with the cell membrane, since GAP 43(Ala3Ala4), a mutant that failed to associate with the cell cortex, had no morphogenetic activity. Two GAP-43 phosphorylation mutants (Ser42 to Ala42 preventing and Ser42 to Asp42 mimicking phosphorylation by PKC) modulated the effects of GAP-43 in opposite ways. Cells expressing GAP-43(Asp42) spread extensively and displayed large and irregular membranous extensions with little filopodia, whereas GAP-43(Ala42) produced small, poorly spreading cells with numerous short filopodia. Therefore, GAP-43 influences cell surface behavior and phosphorylation modulates its activity. The presence of GAP-43 in growing axons and developing nerve termini may affect the behavior of their actin-containing cortical cytoskeleton in a regulatable manner. PMID- 8421063 TI - Identification and characterization of a tumor cell receptor for CSVTCG, a thrombospondin adhesive domain. AB - We have previously shown that peptides derived from the thrombospondin sequence, CSVTCG, promoted tumor cell adhesion. To further investigate this observation, the CSVTCG-tumor cell adhesion receptor from A549 human lung adenocarcinoma cells was isolated and characterized. A single protein peak was isolated by CSVTCG affinity chromatography which also analyzed as a single peak by anion exchange chromatography. The purified protein had a pI of 4.7 and analyzed on SDS-gels as a single band of M(r) = 50,000 under nonreducing conditions and as two protein bands of M(r) = 50,000, and 60,000 under reducing conditions. Purified CSVTCG binding protein (CBP) bound either CSVTCG- or TSP-Sepharose but showed little interaction with either VCTGSC- or BSA-Sepharose. CBP was cell surface exposed. CSVTCG derivatized with [125I] Bolton-Hunter reagent was taken up by cells in a dose-dependent manner and the cell association was inhibited with a monospecific polyclonal anti-CBP antibody. Examination of the cell proteins crosslinked to labeled CSVTCG by SDS-gel electrophoresis revealed one band that comigrated with purified CPB. Using an in vitro binding assay, purified CBP bound mannose, galactose, and glucosamine-specific lectins. CBP bound TSP saturably and reversibly. The binding was Ca+2/Mg+2 ion dependent and inhibited with fluid phase TSP and anti-CBP. Little or no binding was observed on BSA, fibronectin, GRGES, and GRGDS. Heparin, but not lactose, inhibited binding. Anti-CBP IgG and anti-CSVTCG peptide IgG inhibited A549 cell spreading and adhesion on TSP but not on fibronectin and laminin. These results indicate that CBP and the CSVTCG peptide domain of TSP can mediate TSP-promoted tumor cell adhesion. PMID- 8421064 TI - Interaction of integrins alpha 3 beta 1 and alpha 2 beta 1: potential role in keratinocyte intercellular adhesion. AB - The colocalization of integrins alpha 2 beta 1 and alpha 3 beta 1 at intercellular contact sites of keratinocytes in culture and in epidermis suggests that these integrins may mediate intercellular adhesion (ICA). P1B5, an anti alpha 3 beta 1 mAb previously reported to inhibit keratinocyte adhesion to epiligrin, was also found to induce ICA. Evidence that P1B5-induced ICA was mediated by alpha 2 beta 1 and alpha 3 beta 1 was obtained using both ICA assays and assays with purified, mAb-immobilized integrins. Selective binding of alpha 2 beta 1-coated beads to epidermal cells or plate-bound alpha 3 beta 1 was observed. This binding was inhibited by mAbs to integrin alpha 3, alpha 2, or beta 1 subunits and could be stimulated by P1B5. We also demonstrate a selective and inhibitable interaction between affinity-purified integrins alpha 2 beta 1 and alpha 3 beta 1. Finally, we show that expression of alpha 2 beta 1 by CHO fibroblasts results in the acquisition of collagen and alpha 3 beta 1 binding. Binding to both of these ligands is inhibited by P1H5, an anti-alpha 2 beta 1 specific mAb. Results of these in vitro experiments suggest that integrins alpha 2 beta 1 and alpha 3 beta 1 can interact and may do so to mediate ICA in vivo. Thus, alpha 3 beta 1 mediates keratinocyte adhesion to epiligrin and plays a second role in ICA via alpha 2 beta 1. PMID- 8421065 TI - Multiple functional forms of the integrin VLA-2 can be derived from a single alpha 2 cDNA clone: interconversion of forms induced by an anti-beta 1 antibody. AB - The integrin VLA-2 was previously found to bind to either collagen alone, or collagen plus laminin, but the mechanism for this cell-specific functional difference was unknown. Here we transfected VLA-2 alpha 2 subunit cDNA into K562 cells and obtained VLA-2 (called Form-O) which bound to neither collagen nor laminin. We then used a Matrigel selection procedure to enrich for a minor subpopulation of K562 cells stably expressing a form of VLA-2 (Form-C) that bound to collagen but not laminin. In contrast, the same alpha 2 cDNA transfected into RD cells yielded VLA-2 (Form-CL) which bound to both collagen and laminin. These Form-O, -C, and -CL activities were stably expressed during extended cell culture, and could not be qualitatively altered by adding phorbol esters or by exchaning the resident divalent cations. However, addition of stimulatory anti beta 1 antibodies (TS2/16, A-1A5) rapidly converted VLA-2 Form-O and Form-C into Form-CL. Anti-beta 1 antibody stimulation of VLA-2 activity was observed not only on whole cells, but also with solubilized receptors. These results suggest (a) that the ligand binding specificity of VLA-2 can be determined by its cellular environment, rather than by variations in the primary sequence of the alpha 2 subunit, (b) that stably inactive or partly active VLA-2 can be rapidly converted to a fully active form through conformational changes initiated at a nonligand binding site on the beta 1 subunit, and (c) that the mechanisms for VLA-2 stimulation by phorbol ester and by antibody are quite distinct, because the latter does not require an intact cell. PMID- 8421066 TI - Structure and binding properties of collagen type XIV isolated from human placenta. AB - Collagen XIV was isolated from neutral salt extracts of human placenta and purified by several chromatographic steps including affinity binding to heparin. The same procedures also led to the purification of a tissue form of fibronectin. Collagen XIV was demonstrated by partial sequence analysis of its Col1 and Col2 domains and by electron microscopy to be a disulphide-linked molecule with a characteristic cross-shape. The individual chains had a size of approximately 210 kD, which was reduced to approximately 180 kD (domain NC3) after treatment with bacterial collagenase. Specific antibodies mainly to NC3 epitopes were obtained by affinity chromatography and used in tissue and cell analyses by immunoblotting and radioimmunoassays. Two sequences from NC3 were identified on fragments obtained after trypsin cleavage. They were identical to cDNA-derived sequences of undulin, a noncollagenous extracellular matrix protein. This suggests that collagen XIV and undulin may be different splice variants from the same gene. Heparin binding was confirmed in ligand assays with a large basement membrane heparan sulphate proteoglycan. This binding could be inhibited by heparin and heparan sulphate but not by chondroitin sulphate. In addition, collagen XIV bound to the triple helical domain of collagen VI. The interactions with heparin sulphate proteoglycan and collagen VI were not shared by the NC3 domain, or by reduced and alkylated collagen XIV. No or only low binding was observed for collagens I-V, pN-collagens I and III, and several noncollagenous matrix proteins, including laminin, recombinant nidogen, BM-40/osteonectin, plasma and tissue fibronectin, vitronectin, and von Willebrand factor. Insignificant activity was also shown in cell attachment assays with nine established cell lines. PMID- 8421067 TI - Old and new facts about perinatal brain development. PMID- 8421068 TI - Parenting provided by adults with mental retardation. AB - Studies assessing the quality of parenting provided by adults with mental retardation present conflicting conclusions. Some consider the majority to be doing reasonably well, whilst others report frequently unsatisfactory caretaking. There are a number of reasons for such different views. First, inconsistent selection criteria make it hard to compare across studies. In particular, sample composition will be influenced by the recruitment source. For example, if parents have been chosen from voluntary educational programmes a rather different picture is likely to be found than if they have been selected from individuals known to, or referred by, statutory agencies. On the whole, authors working with subjects from the former source have been rather more optimistic than those working with parents referred because there were already serious concerns about parenting difficulties or about delayed child development. Secondly, the majority of studies have used poorly defined global measures of parenting, with variable criteria of what constitutes adequate care. Some have concentrated on physical care and hygiene, whilst others have looked for the presence of affection and warmth. A child's reception into care as the sole measure of the quality of parenting is an unsatisfactory criterion because parental retardation has itself occasionally been used as the basis for removal of a child into care, even in the absence of other evidence of neglect or abuse. Thirdly, methodological flaws are found in studies that have used observational assessments of parenting. Such studies have suggested mothers with mental retardation tend to lack interactive skills (such as high levels of praise and imitation, and low restrictiveness) which are known to be associated with optimal child development. Control groups have often not been matched on social and other variables which might be expected to exert a significant influence upon parenting practices. In addition, the generalisability of these observational studies is open to question as parenting style has been adduced from brief play sessions, lasting at the most 10 minutes. The extent to which mothers with mental retardation play spontaneously with their children at home in a stimulating and age-appropriate fashion has not been measured. With regard to the evidence on abuse and neglect, questions have been raised about whether the children of parents with mental retardation are at increased risk. This problem has rarely been addressed in a methodologically satisfactory way. First, most studies have drawn their samples from referrals to medical or psychiatric departments. Secondly, as indicated earlier, reception into care cannot by itself be taken to be an indicator of child abuse.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8421069 TI - Controversy, theory and social context in contemporary day care research. PMID- 8421070 TI - Structured interviews for assessing children. AB - Structured interviews provide a valuable means of obtaining and quantifying information about the mental status of children. This review indicates that children can reliably self-report and that the information they provide can concur with the opinion of adults knowledgeable about them. However, considerably more research is warranted before it may be assumed that these interviews are adequately reliable and valid. In general, it appears that the task of documenting the psychometric soundness of these interviews has not been taken seriously, as if content validity were sufficient. For example, except for the CAS and the DISC, there has been little effort to study contrast group validity (i.e. whether the interview even differentiates "known groups"). More specifically, review of the reliability and validity data relevant to DSM-III-R diagnoses provides support for the CAS, DICA, ISC and K-SADS, with the validity data for the DICA being weaker than for the others. One limitation of these data for the DICA, ISC and K-SADS is that the diagnoses were clinician-generated, rather than algorithm-generated. Unfortunately the processes for generating clinical diagnoses were not specified, except for criterion reference to DSM-III R. The findings for the DISC and the DISC-R are notably weak. There is no evidence for DISC reliability, except for adolescents, and the validity studies have demonstrated only weak relationships. There has been limited study of the psychometric properties of symptom scales. In fact, for two interviews (i.e. DICA and ISC), there are no data available. Reliability for the DISC scales is adequate only for adolescents. Psychometric data have been generated for the CAS and the K-SADS, with considerably more studies conducted with the CAS. The relative paucity of interest in scale scores is striking given that they provide a continuous variable which can indicate extent of symptoms. Other measures of mental status, besides presence/absence of diagnosis, will become increasingly important as research in child psychopathology progresses toward more sophisticated studies (i.e. treatment effects, risk factors). These interviews are labor intensive and costly to the researcher as well as time-consuming and tedious for the children and parents. Given this commitment, researchers should invest in developing other ways of exploiting the richness of the data generated. An example is the CAS "content" scales, which generate scores reflecting on the child's functioning in various areas (e.g. school, friends, family). As the evolution of these interviews continues, it will be important to remain attentive to the developmental limitations of children.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8421071 TI - Recent advances in selected aspects of adolescent development. PMID- 8421072 TI - Effects of calcium restriction on metabolic characteristics of premenopausal women. AB - We studied 28 healthy premenopausal women before and after manipulating Ca intake, and then in response to a hypercalcemic challenge. Women with low Ca intakes at entry (< 17.5 mmol/day) were restricted to about 5 mmol/day (low-Ca), and those with higher self-selected intakes were supplemented to about 70 mmol/d (high-Ca). After 8 weeks on these regimens, more bone resorption was occurring among the low-Ca women, as evidenced by their higher values for immunoreactive PTH and urine hydroxyproline. There was a 7-fold range in 24-h urine Ca among the low-Ca women, with upper values equivalent to about 80% of intake. In response to induced hypercalcemia (0.5 mmol Ca/kg lean body mass, infused over 4 h), the low Ca group had greater increases in serum Ca (1.31 vs. 1.05 mmol/L, P < 0.05) and reached a marginally higher peak (3.64 vs. 3.50 mmol/L, P < 0.1). Despite these greater calcemic responses, the low-Ca women excreted a smaller fraction of the infused Ca (44.3 vs. 62.2%, P < 0.02). Furthermore, preinfusion urine Ca was directly correlated with excretion of infused Ca in the low-Ca women, but not in the high-Ca women. Preinfusion differences between groups in immunoreactive PTH and urine hydroxyproline tended to reappear within 2 days. Our findings show that there are detectable differences in the Ca regulatory system between subjects at practical extremes of Ca intake, and that these differences persist through exposure to a temporary Ca surfeit. Ca-restricted women have higher levels of PTH and of bone remodeling activity. There is considerable variation in the apparent capacity to conserve Ca among women with low intakes. None of our subjects had high-P diets. Thus, our findings show that Ca restriction can evoke a persistent PTH response in the absence of a high P intake. PMID- 8421073 TI - Low aromatase activity in microsomes from complete hydatidiform mole. AB - The complete hydatidiform mole (CHM) is characterized by the presence of aberrant placenta, with hyperplasia of cyto- and syncytiotrophoblasts and the absence of maternal genetic information. Steroidogenesis in this condition is, thus, of special interest. In this study we investigated the kinetic parameters of aromatase in microsomes from CHM compared with those in normal early placenta (NEP). The enzyme activity was determined by measuring the conversion of [3H] testosterone to [3H]estradiol plus [3H]estrone. The Km value for testosterone was 33 nmol/L in CHM and 17 nmol/L in NEP of similar gestational ages. Aminoglutethimide, a nonsteroidal inhibitor, decreased in a dose-dependent manner and with the same potency the aromatization of testosterone in both tissues (ID50, 2 vs. 1 mumol/L in CHM and NEP, respectively). These results suggest that the enzymes from the two sources are kinetically similar. However, the enzyme efficiency, expressed as the maximum velocity/Km ratio, was 17-fold lower in CHM than in NEP tissue (1.22/33 vs. 10.68/17 min/mg.mL). These findings suggest that in molar pregnancy the decreased capacity of trophoblast tissue for the formation of estrogen could increase the testosterone concentration inside the molar vesicle, which, in turn, as we previously reported, inhibits progesterone formation. All of these data could provide an explanation for the low circulating level of progesterone, which may directly or indirectly affect the spontaneous expulsion of this aberrant tissue in the second trimester of pregnancy. PMID- 8421074 TI - Prenatal dexamethasone treatment in pregnancies at risk for congenital adrenal hyperplasia due to 21-hydroxylase deficiency: effect on midgestational amniotic fluid steroid levels. AB - Prenatal diagnosis of congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency by amniotic fluid (AF) steroid analysis is not possible in those cases in which prenatal dexamethasone (DEX) therapy is initiated to prevent virilization of female CAH fetuses because AF steroid levels are suppressed if DEX therapy is continued beyond amniocentesis (AC). In order to use AF steroids for prenatal diagnosis, it is necessary to discontinue DEX therapy for 5 days before AC. To study the effects of this interruption on AF steroid levels, we measured levels of aldosterone, corticosterone, 11-deoxycorticosterone, progesterone, 17-hydroxyprogesterone (170HP), 11-deoxycortisol, cortisol, and cortisone as well as androstenedione (delta 4-A) in AF samples (16-18 weeks) obtained from 25 pregnancies at risk for CAH treated with Dex (daily dosage: 1.0 1.5 mg). The prenatal diagnosis of 14 normal fetuses and 11 affected CAH fetuses was postnatally confirmed in all cases. Additionally, steroid levels were measured in AF samples (16-18 weeks) from 8 untreated CAH fetuses and in 19 AF samples (weeks 16-20) obtained in normal pregnancies. In 17/19 prenatally diagnosed CAH fetuses, the affected sibs had the salt wasting (SW)-form, in 2 cases the simple virilizing (SV)-form. All steroids were measured by RIA after extraction and Sephadex LH-20 chromatography. AF levels of aldosterone, corticosterone, deoxycorticosterone, progesterone, cortisol, cortisone, and 11 deoxycortisol were not different between CAH fetuses, prenatally DEX-treated normal fetuses and untreated controls. The 170HP-levels of the CAH-SW-fetuses (range: 19.9-59.8 mmol/L) were clearly above the normal range (3.74-11.6), but normal in the SV-fetuses (10.9, 11.5), whereas delta-4 A-levels (normal range: 0.87-5.13 mmol/L) were elevated both in the SW-(range: 6.53-37.6) and the SV-form (9.37,6.25) of CAH. 170HP and delta-4 A levels of prenatally DEX-treated pregnancies with normal fetuses were not different from levels found in normal pregnancies. Mean 170HP and delta-4 A AF steroid levels of prenatally DEX-treated CAH-pregnancies were slightly lower (NS) than levels of untreated CAH-pregnancies (170HP: 30.5 vs. 40.7; delta-4 A: 15.8 vs. 21.1). 170HP levels are elevated in the SW-form of CAH, but not in the SV-form. However, with the combination of 170HP and delta-4 A levels it is possible to diagnose prenatally both forms. There is no rebound phenomenon of AF steroid levels if DEX therapy is interrupted 5 days before amniocentesis. PMID- 8421075 TI - Lipoprotein(a) and apolipoprotein B plasma concentrations in hypothyroid, euthyroid, and hyperthyroid subjects. AB - Overt hypothyroidism is associated with premature coronary artery disease, and this is assumed to be due to a deteriorated metabolism of atherogenic lipoproteins. The effect of thyroid status on plasma concentrations of lipoprotein(a) [Lp(a)], a recently recognized highly atherogenic lipoprotein in man, is unknown. In a cross-sectional study, plasma Lp(a) concentrations were higher in overtly hypothyroid subjects [255 +/- 28 (+/- SD) mg/L; n = 19] and lower in hyperthyroid subjects (75 +/- 28 mg/L; n = 27) compared to those in 54 euthyroid subjects (150 +/- 36 mg/L) and a reference population of local blood bank donors (155 +/- 31 mg/L; n = 114). These findings were confirmed in a follow up study of 19 hypothyroid and 8 hyperthyroid individuals. In the hypothyroid subjects, initial levo-T4 substitution therapy (25 micrograms daily) caused a 55% decrease in plasma Lp(a) concentrations and a 27% decrease in total plasma apolipoprotein B (apo B). Good agreement was found between the decrease in Lp(a) and apo B at a normal free T4 index. Follow-up of 8 hyperthyroid subjects revealed that their plasma Lp(a) and apo B concentrations significantly increased with return of euthyroidism. In conclusion, good agreement was found between the direction and magnitude of the responses of apo B and Lp(a) to changes in thyroid status. The following findings suggest that different thyroid hormone-dependent mechanisms modulate plasma Lp(a) concentrations in man, in part analogous to modulation of apo B: 1) impaired catabolism in the hypothyroid state, and 2) a combination of suppressed secretion of apoB and Lp(a) with increased catabolism in hyperthyroid subjects. Increased plasma Lp(a) concentrations may contribute to the increased risk of premature coronary artery disease in the hypothyroid state. PMID- 8421076 TI - Autocrine stimulation of interleukin-1 in the growth of human thyroid carcinoma cell line NIM 1. AB - We reported first in this study that human thyroid cell line NIM 1 established from a patient with papillary adenocarcinoma of the thyroid associated with hypercalcemia and peripheral neutrocytosis produced interleukin (IL)-1 alpha and IL-1 beta in the culture supernatant and cell lysate as detected by murine thymocyte proliferative response and enzyme-linked immunosorbent assay. Production of IL-1 alpha and IL-1 beta was further confirmed by the demonstration of IL-1 alpha and IL-1 beta messenger ribonucleic acid expression with Northern blot hybridization analysis. The in vitro growth of NIM 1 cells was inhibited by the addition of anti-IL-1 alpha and IL-1 beta antibody. The growth of NIM 1 cells was further enhanced by the addition of recombinant human IL-1 alpha and IL-1 beta, whereas this enhancement was also inhibited by the addition of anti-IL-1 antibody. IL-1 receptors were expressed on NIM 1 cells. These results suggest that IL-1 plays a regulatory role in the growth of NIM 1 cells by an autocrine mechanism. PMID- 8421077 TI - Continuous subcutaneous infusions of growth hormone (GH) releasing hormone 1-44 for 14 days increase GH and insulin-like growth factor-I levels in old men. AB - Twice daily sc injections of GHRH increase serum GH and IGF-I levels in healthy old men to values like those of untreated young men by producing high amplitude GH peaks after the injections. In the present study, we measured baseline insulin like growth factor-I (IGF-I) 24-h profiles of GH release, and responses to GHRH stimulation tests in healthy young and old men. Old men were then given, in random order, 1 and 2 mg continuous sc GHRH 1-44 infusions daily for 14 days with an intervening 14-day treatment-free period. The study protocol was repeated on day 14 of each treatment. At baseline, mean duration of GH peaks (P < 0.005) and IGF-I levels (P < 0.0001) were lower in old men. Both doses increased (vs. old basal) mean 24-h GH, integrated area under the GH curve, and GH peak number (P < 0.05), as well as serum IGF-I (P < 0.001). Interpeak GH levels also increased during low (P < 0.01) and high (P < 0.05) dose treatment. Significant increases in mean GH and integrated area under the GH curve occurred only during the day (0800-2000 h) during both low (P < 0.01) and high (P < 0.05) dose treatment. Treatment also decreased nocturnal peak GH amplitude and duration. GH responses to GHRH stimulation tests did not differ with age at baseline, or in old men after treatment. Thus, continuous, short-term sc administration of GHRH to healthy old men restores subnormal GH secretion and IGF-I levels by increasing GH peak frequency and interpeak secretion, particularly during the day. PMID- 8421078 TI - Progression of uremic hyperparathyroidism involves allelic loss on chromosome 11. AB - In occasional cases of secondary hyperparathyroidism, long term stimulation of the parathyroid glands leads from compensatory to autonomous hyperfunction, and thus, hypercalcemia develops. This clinical entity, named tertiary hyperparathyroidism, is possibly due to the formation of an adenoma in one of the hyperplastic glands. Previous studies have shown that parathyroid adenomas may arise with allelic loss on chromosome 11. We tested for allelic loss at several loci on chromosome 11 in 12 enlarged parathyroid glands from 6 uremic patients and found loss of heterozygosity in 2 of the glands from 2 different patients with higher serum calcium levels (11.3 +/- 0.29 vs. 9.8 +/- 0.28 mg/dL; P < 0.004) and, therefore, ascribable to the so-called tertiary hyperparathyroidism. The 2 glands with allelic loss were significantly greater in mass than those without loss (3.42 +/- 0.37 vs. 1.60 +/- 0.54 g; P < 0.001). These data offer new evidence that autonomous parathyroid proliferation in uremic patients can develop through overgrowth by a monoclonal tumor, presumably with inactivation of a tumor suppressor gene(s) on chromosome 11. PMID- 8421079 TI - Clinical review 41: Current therapy for osteoporosis. PMID- 8421080 TI - A radioimmunoassay for measurement of thyroxine sulfate. AB - A highly sensitive, specific, and reproducible RIA has been developed to measure T4 sulfate (T4S) in ethanol extracts of serum. rT3 sulfate (rT3S) cross-reacted 7.1%, and T3S cross-reacted 0.59% in the RIA; T4, T3, rT3, and 3,3' diiodothyronine cross-reacted 0.004% or less. The recovery of nonradioactive T4S added to serum averaged 95%. The detection threshold of the RIA was 18 pmol/L. The coefficient of variation averaged 6.9% within an assay and 12% between assays. T4S was bound by T4-binding globulin and albumin in serum. The free fraction of T4S in four normal sera averaged 0.06% compared to a value of 0.03% for T4 (P < 0.001). The serum concentration of T4S was (mean +/- SE) 19 +/- 1.2 pmol/L in normal subjects, 33 +/- 10 in hyperthyroid patients with Graves' disease, 42 +/- 15 in hypothyroid patients, 34 +/- 6.9 in patients with systemic nonthyroidal illnesses, 21 +/- 4.3 in pregnant women at 15-40 weeks gestation, and 245 +/- 26 in cord blood sera of newborns; the value in the newborn was significantly different from normal (P < 0.001). The mean concentration of T4S in amniotic fluid samples at 15-38 weeks gestation was 106 +/- 22 pmol/L (cf. normal adults; P < 0.001). Administration of sodium ipodate (Oragrafin; 3 g, orally) to hyperthyroid patients was associated with a transient increase in serum T4S. The T4S content of the thyroid gland was less than 1/4000th that of T4. We conclude that 1) T4S is a normal component of human serum, and its levels are markedly increased in newborn serum and amniotic fluid; and 2) the sulfation pathway plays an important role in the metabolism of T4 in man. PMID- 8421081 TI - Treatment of familial male precocious puberty with spironolactone, testolactone, and deslorelin. AB - Combined antiandrogen (spironolactone) and aromatase inhibitor (testolactone) are effective for the short term treatment of familial male precocious puberty. During this therapy, plasma testosterone levels remain in the adult range, since spironolactone blocks the testosterone receptor without significantly affecting plasma testosterone levels. After our initial 18-month pilot study, we continued to treat eight boys with the combined therapy for 2.0-4.2 yr. During this time all boys exhibited a pubertal rise in gonadotropin secretion and a diminishing response to treatment, which was manifested by the recurrence of clinical features of puberty and an increase in the bone maturation rate (P < 0.05). Addition of the LHRH agonist deslorelin (4 micrograms/kg.day, sc) to the combined therapy decreased peak LH, plasma testosterone, bone maturation rate, and growth velocity (P < 0.05) over the next year. We conclude that the rise in gonadotropin levels during central activation of hypothalamic LHRH secretion in boys with familial male precocious puberty causes a partial escape from the combined effect of spironolactone and testolactone. The addition of deslorelin to the combined therapy appears to restore the control of puberty in this setting. PMID- 8421082 TI - Endothelin binding to receptors and endothelin production by human thyroid follicular cells: effects of transforming growth factor-beta and thyrotropin. AB - Measurable endothelin (ET) was found in serum-free medium of cultured primary thyroid cells derived from human thyroid tissues after 3 days incubation at levels ranging from undetectable to 35 fmol/100,000 cells. Out of 12 thyroid glands studied, 2 responded to transforming growth factor-beta (TGF-beta) treatment with increased ET secretion into medium. Detailed study of cells derived from one of these thyroids showed TGF-beta at 1 ng/ml stimulated ET secretion from 13.7 to 33.3 fmol/100,000 cells after 3 days incubation. Although TSH alone did not significantly modulate ET release into medium, TSH enhanced the stimulatory effect of TGF-beta. A combination of TSH at 1 mU/ml and TGF-beta at 1 ng/ml stimulated ET secretion to 63.8 fmol/100,000 cells after 3 days incubation. Immunostaining studies demonstrated the presence of intracellular immunoreactive ET, largely localized in perinuclear regions, which was greatly stimulated by TSH but not by TGF-beta. These observations suggest that TSH may stimulate only ET synthesis, whereas TGF-beta may stimulate both synthesis and secretion. Binding of [125I]ET-1 to receptor on thyroid cells was dose-dependently stimulated by TGF beta (0-10 ng/ml) pretreatment for 3 days. Scatchard analysis of competitive binding data from TGF-beta (1 ng/ml)-treated cells indicated that increased binding was the result of increased receptor number and not increased receptor affinity. TSH (0-10 mU/ml), though not as potent as TGF-beta, also dose dependently stimulated ET binding. Treatment of thyrocytes with 1 mU/ml TSH for 3 days did not significantly affect ET receptor number or binding affinity. These results illuminate aspects of a possible autocrine regulation of ET in the thyroid gland. PMID- 8421083 TI - The serum growth hormone-binding protein is reduced in young patients with insulin-dependent diabetes mellitus. AB - Despite elevated serum concentrations of GH, longitudinal growth is stunted in a considerable number of children and adolescents with insulin-dependent diabetes mellitus (IDDM). To elucidate, whether reduced peripheral action of GH contributes to this phenomenon, GH-binding protein (GH-BP) activity was measured in 117 children and adolescents with IDDM (mean age 14.6 yr, range 4.5-28 yr) and 132 healthy controls (13.1 yr, 6.3-26 yr). Serum was incubated with 125I-GH, then chromatographed on a Sephacryl S200 column (1.8.100 cm), apparent binding of 125I GH to GH-BP was corrected for the amount of endogenous GH present in the sample. GH-BP activity was significantly lower in IDDM patients, with a corrected binding of 16.8 +/- 0.6% compared to 21.3 +/- 0.7% in control children (mean +/- SE; P < 0.0001, Wilcoxon-test). Previous studies demonstrated that GH-BP is increased in healthy overweight children. In contrast, in IDDM children, GH-BP was reduced despite a moderate degree of overweight (z-score for weight: +0.94 +/- 0.12; mean +/- SE). Reduced serum GH-BP activity in IDDM children is further accentuated when compared to healthy children with a similar degree of overweight (22.8 +/- 0.5%; n = 44). Based on this novel finding, we conclude that decreased GH receptor density may explain reduced growth velocity despite increased secretion of GH in some IDDM children. PMID- 8421084 TI - Effect of growth hormone (GH)-releasing hormone (GHRH), atropine, pyridostigmine, or hypoglycemia on GHRP-6-induced GH secretion in man. AB - His-DTrp-Ala-Trp-DPhe-Lys-NH2 (GHRP-6) is a synthetic compound that releases GH in a dose-related and specific manner in several species, including man. To further characterize the effects and mechanism of action of GHRP-6 on GH secretion, we assessed in normal man plasma GH responses to that hexapeptide 1) alone and in combination with exogenous GH-releasing hormone (GHRH) administration, 2) in a state of high endogenous somatostatinergic tone after atropine administration, and 3) in a state of low endogenous somatostatinergic tone induced by the cholinergic receptor agonist drug pyridostigmine or after insulin-induced hypoglycemia. We found a similar increase in plasma GH levels after the administration of either GHRP-6 (1 microgram/kg) or GHRH (1 microgram/kg); the areas under the curve (AUC) were (mean +/- SEM) 973 +/- 181 and 821 +/- 139, respectively. After combined GHRP-6 and GHRH administration, GH responses were considerably greater than those after either compound alone (4412 +/- 842; P < 0.01). Administration of the cholinergic receptor antagonist atropine (1 mg, im) completely prevented the GH responses to GHRP-6 (area under the curve, 103 +/- 14 vs. 815 +/- 156, respectively). On the other hand, pyridostigmine, a cholinergic agonist, slightly increased GH responses to GHRP-6 (P < 0.01 when comparing the AUC after pyridostigmine administration of 1571 +/- 151 and the AUC after administration of GHRP-6 alone of 815 +/- 156). Finally, combined GHRP-6 and insulin administration induced a much greater increase in plasma GH levels (AUC, 4047 +/- 327) than insulin alone (1747 +/- 229; P < 0.05) or GHRP-6 alone (1248 +/- 376; P < 0.05). Our results lend support to the view that GHRP-6-induced GH secretion is exerted through a non-GHRH-dependent mechanism. Furthermore, the fact that enhancement of somatostatinergic tone with atropine completely prevented the GH responses to GHRP-6, while pyridostigmine and insulin-induced hypoglycemia, which increased plasma GH levels by inhibiting hypothalamic somatostatin release, increased the same response suggest that although GHRP-6-induced GH secretion is dependent on the endogenous somatostatinergic tone, the stimulatory effect of GHRP-6 on plasma GH levels is not mediated by a change in hypothalamic somatostatinergic tone. PMID- 8421085 TI - Genetic basis of endocrine disease. 4. The spectrum of mutations in the androgen receptor gene that causes androgen resistance. AB - Mutations in the androgen receptor gene cause phenotypic abnormalities of male sexual development that range from a female phenotype (complete testicular feminization) to that of undervirilized or infertile men. Using the tools of molecular biology, we have analyzed androgen receptor gene mutations in 31 unrelated subjects with androgen resistance syndromes. Most of the defects are due to nucleotide changes that cause premature termination codons or single amino acid substitutions within the open reading frame encoding the androgen receptor, and the majority of these substitutions are localized in three regions of the androgen receptor: the DNA-binding domain and two segments of the androgen binding domain. Less frequently, partial or complete gene deletions have been identified. Functional studies and immunoblot assays of the androgen receptors in patients with androgen resistance indicate that in most cases the phenotypic abnormalities are the result of impairment of receptor function or decreases in receptor abundance or both. PMID- 8421086 TI - Differences in postprandial lipemia between patients with normal glucose tolerance and noninsulin-dependent diabetes mellitus. AB - In this paper we have compared the postprandial increase in triglyceride (TG) rich lipoproteins of intestinal origin in 10 patients with noninsulin-dependent diabetes mellitus (NIDDM) and 10 subjects with normal glucose tolerance. The two groups were matched for age, sex distribution, body mass index, and plasma TG concentration. Breakfast was consumed at 0800 h and lunch at 1200 h, at which time vitamin A was also administered. Blood was sampled frequently from 1200 h to 2400 h, and measurements made of glucose, insulin, and TG concentrations. Furthermore, the retinyl palmitate (RP) content of plasma, the Sf > 400 lipoprotein fraction, and the Sf 20-400 lipoprotein fraction was also determined, and differences compared by two-way analysis of variance. Fasting and postprandial (from 1200 h to 2400 h) TG concentrations in the plasma and the two lipoprotein fractions were not significantly different in normal subjects and patients with NIDDM. In addition, the postprandial RP concentration of the two groups was not different in the chylomicron containing Sf > 400 lipoprotein fraction. However, the postprandial Sf 20-400 RP concentration was significantly higher (P < 0.001) in patients with NIDDM, estimated as hourly values over time, peak value, or total integrated response area. Significant correlation coefficients (r = 0.60-0.75, P < 0.08 < 0.02) were seen in patients with NIDDM between the total integrated insulin response and both the TG and RP responses in the Sf > 400 and Sf 20-400 fractions. In addition, fasting high density lipoprotein-cholesterol concentration in patients with NIDDM was significantly correlated with the postprandial TG response in the Sf > 400 (r = -0.64, P < 0.05) and the Sf 20-400 (r = -0.68, P < 0.05) lipoprotein fractions. In summary, the postprandial RP concentration in the Sf 20-400 lipoprotein fraction was higher than normal in patients with NIDDM. In addition, associations have been defined in patients with NIDDM between postprandial insulin response, fasting TG and high density lipoprotein-cholesterol concentrations, and magnitude of postprandial increase in TG-rich lipoproteins of intestinal origin. PMID- 8421087 TI - Effects of nitrendipine on glucose tolerance and serum insulin and dehydroepiandrosterone sulfate levels in insulin-resistant obese and hypertensive men. AB - To determine whether the calcium channel blocker nitrendipine improves glucose tolerance, lowers circulating insulin, and raises serum dehydroepiandrosterone sulfate (DHEA-S) levels in insulin-resistant men, a total of 15 obese and hypertensive men were enrolled in a single blind, placebo-controlled study. A nitrendipine group (n = 8) and a placebo group (n = 7) were studied before and after treatment with either nitrendipine (10 mg) or a placebo capsule, twice daily for 7 days, by determining serum insulin, glucose, and DHEA-S levels in the fasting state and during an oral glucose tolerance test. Nitrendipine treatment 1) lowered fasting serum insulin from 265 +/- 24 to 194 +/- 22 pmol/L (P < 0.01) without changing fasting serum glucose, 2) reduced both the area under the curve for glucose (from 1246 +/- 31 to 1091 +/- 26 mmol/L.min; P < 0.005) and the area under the curve for insulin (from 123.6 +/- 9.4 to 82.9 +/- 10.0 nmol/L.min; P < 0.015) during the oral glucose tolerance test, and 3) increased fasting serum DHEA-S by 63% from 4.21 +/- 0.17 to 6.84 +/- 0.21 mumol/L (P = 0.0001). No change was noted in the placebo group. We conclude that nitrendipine treatment is associated with improved glucose tolerance, reduced fasting and glucose stimulated serum insulin levels, and increased circulating DHEA-S levels. PMID- 8421088 TI - Differences in the low density lipoprotein subfraction profile between oral contraceptive users and controls. AB - To investigate the effect of low dose oral contraceptives on the low density lipoprotein (LDL) subfraction profile, the distribution of the LDL subfraction patterns in 20 premenopausal women on oral contraceptive (OC) therapy and 41 premenopausal women not taking gonadal hormones was studied. The LDL subfraction patterns were identified by density gradient ultracentrifugation and each individual LDL subfraction pattern was characterized by the relative contribution of three major LDL subfractions: light, LDL1; intermediate, LDL2; and dense, LDL3 to total LDL. Serum lipid and lipoprotein levels were similar in OC users and controls, except for significantly higher triglyceride levels in OC users. As for the LDL subfraction patterns, among the OC users the mean relative contribution of dense LDL3 to total LDL was significantly higher than in the controls (32% +/- 8% vs. 26% +/- 13%, P < 0.05), whereas the relative contribution of light LDL1 to total LDL was significantly lower (27% +/- 8% vs. 34% +/- 10%, P < 0.01), indicating a higher prevalence of the more dense LDL subfraction patterns among OC users. Furthermore, the distribution of the LDL subfraction patterns in OC users (27% LDL1, 41% LDL2, and 32% LDL3) resembled that of men (25% LDL1, 43% LDL2, and 33% LDL3, n = 59). Statistical analysis revealed that OC use was significantly associated with a more dense LDL subfraction pattern, characterized by an increased relative contribution of LDL3 (+6%, P < 0.05) and a decreased relative contribution of LDL1 (-6%, P < 0.01), even after correcting for the influence of lipid and lipoprotein levels, which in controls were shown to have a significant relation to LDL3 and LDL1, respectively. So, independent of the lipid and lipoprotein levels, low dose OC alter the composition of LDL to a heavy, dense LDL subfraction profile, which reportedly has been associated with an increased risk of atherosclerosis. PMID- 8421089 TI - Congenital gigantism due to growth hormone-releasing hormone excess and pituitary hyperplasia with adenomatous transformation. AB - The cause of gigantism in most patients is a GH-secreting pituitary tumor. In this report, a case of congenital gigantism due to probable central hypersection of GH-releasing hormone (GHRH) is described. Normal at birth (4.4 kg; 53 cm), our 7-yr-old male patient grew progressively thereafter to attain a height of 182 cm and a weight of 99.4 kg at the time of our evaluation. The markedly increased baseline plasma levels of GH (730 micrograms/L) did not suppress during a standard 3-h oral glucose tolerance test, but did increase 54% after iv infusion of GHRH. Baseline plasma levels of insulin-like growth factor-I, PRL, and immunoreactive GHRH were also markedly increased. Computed imaging of the head showed a large, partially cystic sellar and suprasellar mass. Extensive imaging studies did not localize a potential source of GHRH. Preoperative treatment with octreotide and bromocriptine for 4 months resulted in a 25% reduction of suprasellar tissue mass. The pituitary tissue removed at transsphenoidal and transfrontal operations showed massive somatotroph, lactotroph, and mammosomatotroph hyperplasia. Areas of GH- and PRL-secreting cell adenomatous transformation were also evident. No histological or immunohistochemical evidence of a pituitary source of GHRH was found. The peripheral plasma immunoreactive GHRH concentration remained unaffected by pharmacological and surgical interventions. We suspect that a congenital hypothalamic regulatory defect may be responsible for the GHRH excess in this case. PMID- 8421090 TI - Progestational regulation of human endometrial stromal cell tissue factor expression during decidualization. AB - Decidualized endometrial stromal cells from mid- to late secretory phase and decidual cells from gestational human endometrium displayed prominent immunohistochemical staining for tissue factor, a primary initiator of hemostasis. Consistent with the regulation by progesterone of the decidualization process in vivo, medroxy-progesterone acetate elevated the tissue factor content of primary stromal cell cultures 8-fold over basal values. This increase paralleled the release of immunoreactive PRL, a marker of decidualization. The induced, as well as basal, tissue factor displayed full functional activity and the expected electrophoretic mobility (46 kilodaltons). Moreover, Northern analysis of RNA from cultured stromal cells indicated that medroxyprogesterone acetate increased tissue factor mRNA levels approximately 10-fold relative to control levels. In contrast, cultured stromal cell tissue factor protein content and mRNA levels were unaffected by exogenous estradiol. These findings indicate that enhancement of endometrial stromal cell tissue factor content is associated with progesterone induction of the decidualization process. In humans, trophoblastic invasion of the endometrial vasculature during blastocyst implantation risks hemorrhage. Therefore, increases in perivascular decidual cell tissue factor expression could serve to promote periimplantational endometrial hemostasis. PMID- 8421091 TI - Biochemical characterization and autoradiographic localization of [125I]endothelin-1 binding sites on trophoblast and blood vessels of human placenta. AB - The presence of endothelin binding sites in the human placenta raises the question of the precise localization of these receptors on well defined placental constituents. In order to find an answer to this problem various approaches were used. Specific binding sites for [125I] endothelin-1 (ET-1) were identified on human term placenta, not only on membranes of smooth muscles stem villi vessels, but also on trophoblastic plasma membranes prepared from trophoblast in culture. Scatchard analysis of binding data revealed a single class of high affinity binding sites with Kd values of 26 +/- 4 pmol/L for stem villi vessels and 126 +/ 4 pmol/L for trophoblast in culture, with maximum binding capacities of 681 +/- 61 and 224 +/- 53 fmol/mg protein, respectively. The anatomical localization of these binding sites was determined by in vitro autoradiography. Autoradiograms obtained from placental sections incubated with [125I]ET-1 indicate that [125I]ET 1 high affinity binding sites exist on placental stem villi vessels and on the trophoblastic layer of the villi. The latter localization was also found on autoradiograms of trophoblast in culture. The human placental syncytiotrophoblast is a polarized epithelium with the microvillous membrane, facing maternal blood space and the basal plasma membrane, facing fetal circulation. [125I]ET-1 high affinity binding sites are present on both membranes but the number of binding sites is higher on the basal plasma membrane. These findings lead to the suggestion that ET-1 may be involved in the regulation of the feto-placental circulation and may subserve specific trophoblastic functions. PMID- 8421092 TI - Puberty: when? PMID- 8421093 TI - Serum levels of mullerian inhibiting substance in boys throughout puberty and in the first two years of life. AB - Serum levels of Mullerian inhibiting substance (MIS) have been measured in 91 boys throughout normal pubertal development. MIS levels fell sharply after pubertal stage 1 and were mostly undetectable at pubertal stage 6. The relationship between MIS concentration and pubertal stage was similar when compared with age. Seven patients with precocious puberty and 12 with delayed puberty were also investigated and found to have MIS levels consistent with their degree of pubertal development. Precocious puberty was associated with MIS levels that were abnormally low for age, while delayed puberty resulted in persistence of high MIS levels. Serum MIS levels were also measured in 29 boys less than 2 yr of age undergoing minor surgery. High levels were found throughout this time period, which is consistent with previous reports. MIS levels appear to be inversely related to levels of gonadotropins, steroids, and inhibin, which fall in the first 2 yr of life and rise throughout puberty. PMID- 8421094 TI - The growth hormone and somatomedin axis in short children with osteogenesis imperfecta. AB - Growth deficiency is a cardinal feature of severe osteogenesis imperfecta (OI) and a frequent feature of mild to moderate forms of this disease. We have investigated the status of hormones related to growth in 22 short prepubertal children, 13 males and 9 females, with various types of OI. Ten children had Sillence type III OI, 10 had type IV, and 2 had type I. Evaluation included GRH stimulation, three standard GH provocative tests (arginine-insulin tolerance test, L-dopa), 24-h sampling for measurement of unstimulated GH secretion and a somatomedin-C generation test. None of these children had GH deficiency by standard criteria. We found that 9 OI children had decreased responsiveness to GRH, similar to the GRH response of GH-deficient children. Overall, however, mean 24-h GH values and mean peak GH response to provocative agents of OI children were within the normal range. In the somatomedin generation test, the OI children as a group showed a blunted response, with 13 of 22 having less than a 2-fold stimulation of somatomedin-C by GH. This suggested resistance of the liver and other somatomedin-C secreting tissues to GH. The group with blunted insulin-like growth factor-I response did not correlate significantly with the group with decreased GRH response. To investigate the responsiveness of OI bone to growth stimulation, six OI children with less than average integrated GH secretion were enrolled in a pilot study in which one child received exogenous GH and six received clonidine for at least 6 months. The child treated with exogenous GH and three of six treated with clonidine experienced at least a 4.7 cm/yr increase over their pretreatment growth rates. Growth response could not be predicted from baseline studies. We conclude that abnormalities of the GH-somatomedin axis exist in some children with OI. Administration of GH or clonidine may augment growth rates in OI children; however, the effect of these agents on final stature is unknown. PMID- 8421095 TI - Multiple genetic factors in the heterogeneity of thyroid hormone resistance. AB - Generalized resistance to thyroid hormone (GRTH), a syndrome of inherited tissue hyposensitivity to thyroid hormone, is linked to thyroid hormone receptor (TR) mutations. A typical feature of GRTH is variable severity of organ involvement among families that, surprisingly, does not correlate with the degree of T3 binding impairment of the corresponding in vitro synthesized mutant TRs. Furthermore, variations in the clinical severity among family members harboring identical TR beta mutations have been reported. We compared serum levels of thyroid hormones that maintained a normal TSH in members of a large family with GRTH divided in three groups: Group A, 8 affected subjects with a mutation replacing arginine-320 with a histidine in the T3-binding domain of TR beta; Group B, 11 first degree relatives (sibs and children of affected subjects) with no TR beta mutation; Group C, 16 controls related by marriage. TSH values were not different among the three groups. As expected, total and free T4 and T3, and rT3 levels were significantly higher in Group A vs Groups B and C. However, with the exception of T3, the same tests were also significantly higher in Group B vs Group C. The latter differences are not due to thyroid hormone transport in serum since TBG concentrations were not different. It is postulated that genetic variability of factors that contribute to the action of thyroid hormone modulate the phenotype of GRTH associated with TR beta mutations. PMID- 8421096 TI - Early morning plasma testosterone is an accurate predictor of imminent pubertal development in prepubertal boys. AB - In the management of constitutional delayed growth and/or puberty, there is a need for simple tests which can assess the overall developmental maturity of the hypothalamic-pituitary-testicular axis in clinically prepubertal patients. This would enable the physician to predict the likelihood or otherwise of an individual entering puberty spontaneously within subsequent months. Based on our previous physiological data on the sequential pattern of peripubertal pituitary testicular activation by hypothalamic GnRH, we hypothesized that the nocturnal secretion of testosterone, in response to sleep-entrained LH secretion, may provide a basis for an in vivo bioassay of neuroendocrine sexual maturity. Overnight testosterone secretion by the testis in clinically prepubertal boys was assessed with respect to their subsequent clinical progress, the target being the attainment of testicular volumes of greater than or equal to 4 mL (a clinical landmark when puberty has assuredly begun and virilization will soon follow). Forty-five prepubertal (Tanner stage G1PH1 testicular volume < or = 2 mL) boys aged 10.0-15.3 yr (mean +/- SEM 11.8 +/- 0.2) with short stature had paired plasma T concentration measured at 2000 h and 0800 h the following morning. After the initial assessment, all patients were reviewed clinically at 3-month intervals for a minimum of 21 months (mean 26.0 +/- 1.1, range 21-50 months). During this period, 38 (84.4%) patients received treatment in the form of sc human GH 2-4 IU daily or oxandrolone 2.5 mg daily by mouth to improve short-term growth although this did not have any significant effect on the subsequent timing of pubertal onset. The patients were divided according to whether 1) there was a demonstrable increase in plasma T between 2000 and 0800 h and 2) morning plasma T concentration was less than or greater than or equal to 0.7 nmol/L at their initial assessment. In those with a significant overnight T increment, 58% and 89% achieved testicular volume of greater than or equal to 4 mL after 12 and 21 months, respectively. In contrast, only 12% and 56% of patients who had not shown a T increase went into puberty by these times. In patients who had morning plasma testosterone concentrations greater than or equal to 0.7 nmol/L, 77% entered puberty within 12 months and 100% within 15 months. However, in those with a morning testosterone of less than 0.7 nmol/L, only 12.5% and 25% entered puberty within 12 and 15 months, respectively.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8421097 TI - Corticotropin releasing hormone-binding protein (CRH-BP): plasma levels decrease during the third trimester of normal human pregnancy. AB - In pregnancy, maternal plasma corticotropin releasing hormone (CRH) concentrations rise substantially in the third trimester and fall rapidly post partum. A binding protein (BP) specific for CRH exists in the human circulation which inactivates CRH, thus possibly explaining why maternal ACTH does not rise outside normal limits throughout gestation. We here describe the measurement of CRH-BP directly in plasma during human pregnancy using a radioimmunoassay that is not affected by the presence of the high plasma levels of CRH that occur at this time. In 119 healthy non-pregnant individuals, mean CRH-BP levels were 4.46 nmol/L +/- 1.0 (SD), with a wide range of 1.81-7.24 nmol/L. Plasma CRH-BP in 34 pregnant women randomly sampled during the first and second trimesters also averaged 4.46 nmol/L +/- 1.54, with individual values ranging from 1.59-7.51 nmol/L and there was no correlation of CRH-BP levels with gestational age. In a group of 14 women sampled sequentially throughout the third trimester, plasma CRH BP averaged 4.56 nmol/L +/- 1.70 at 30-35 weeks gestation and fell dramatically to 1.84 nmol/L +/- 0.43 at weeks 38-40 (P < 0.001). The post partum recovery in CRH-BP levels occurred within 48 hours of delivery. These results indicate that there is an increase in the availability of free, potentially bioactive CRH at term to stimulate the release of ACTH from the maternal pituitary and/or to act at a peripheral, non-pituitary CRH receptor(s). PMID- 8421098 TI - Pope Joan: a recognizable syndrome. PMID- 8421099 TI - Prospective study of the long-term effects of somatostatin analog (octreotide) on gallbladder function and gallstone formation in Chinese acromegalic patients. AB - This article reports the changes in gallbladder function examined by ultrasonography in 20 Chinese patients with active acromegaly treated with sc injection of the somatostatin analog octreotide in dosages of 300-1500 micrograms/day for a mean of 24.2 +/- 13.9 months. During treatment with octreotide, 17 patients developed sludge, 10 had gallstones, and 1 developed acute cholecystitis requiring surgery. In all of 7 patients examined acutely, gallbladder contractility was inhibited after a single 100-micrograms injection. In 8 patients followed for 24 weeks, gallbladder contractility remained depressed throughout therapy. After withdrawal of octreotide in 10 patients without gallstones, 8 patients assessed had return of normal gallbladder contractility within 1 month. In 8 of the remaining 10 patients who developed gallstones during treatment, gallbladder contractility normalized in 5 patients (3 of whom has disappearance of their stones within 3 weeks), and remained depressed in 3 (2 of whom had stones present at 6 months). Our results suggest that the suppression of gallbladder contractility is the cause of the successive formation of bile sludge, gallstones, and cholecystitis during octreotide therapy in Chinese acromegalic patients. It is therefore very important to follow the changes of gallbladder function during long-term octreotide therapy of acromegalic patients. PMID- 8421100 TI - Hypoaldosteronism accompanied by normal or elevated mineralocorticosteroid pathway steroid: a marker of adrenal carcinoma. AB - In order to find a biochemical marker to assist the physician in the difficult differential diagnosis between malignant and nonmalignant adrenal tumors, plasma levels of the mineralocorticosteroids (deoxycorticosterone, 18 hydroxydeoxycorticosterone, corticosterone, 18-hydroxycorticosterone, and aldosterone) were determined. The same method (RIA which is preceded by a crucial separation step) was used to measure all these steroids including aldosterone. The subjects included 15 adults presenting various clinical signs of adrenocortical tumors (histopathologically: 6 with adrenal carcinoma, 1 with a history of adrenal carcinoma, 1 with adrenal metastasis from other forms of cancer, 6 with adenoma, and 1 with hyperplasia). The results show that both presurgery and during a recurrence of adrenal carcinoma, hypoaldosteronism occurs which contrasts with the normal or even elevated levels of some aldosterone precursors. In the 7 cases of adrenal cortical carcinoma, this dysfunction of the aldosterone pathway was detected regardless of the impairment of the other steroidogenesis pathways, whereas it was never found with a nonmalignant tumor. Despite the limited number of cases so far available, these findings suggest that detection of abnormalities of the aldosterone pathway, and particularly the detection of hypoaldosteronism by an assay method involving a crucial steroid separating step, could contribute to a differential diagnosis between benign and malignant adrenocortical tumor and between adrenal metastasis and other forms of cancer. PMID- 8421101 TI - Plasma insulin, C-peptide, and proinsulin concentrations in obese and nonobese individuals with varying degrees of glucose tolerance. AB - Conventional immunoassays to quantify insulin concentration do not differentiate between insulin and proinsulin. Thus, previous conclusions as to the relationship between the development of hyperglycemia in patients with noninsulin-dependent diabetes mellitus (NIDDM) and pancreatic insulin secretory function may have been confounded by not being able to determine the contribution made by plasma proinsulin to the putative measurements of plasma insulin concentration in these patients. The current study was initiated to address this issue by making specific measurements of plasma insulin, proinsulin, and C-peptide concentrations in 42 individuals: 14 with normal glucose tolerance, 12 with impaired glucose tolerance (IGT), and 16 with NIDDM. The study population was further subdivided into a nonobese (body mass index, < 30 kg/m2) and an obese (body mass index, > 30 kg/m2) group. Mixed meals were given at 0800, 1200, and 1800 h, and blood was removed at 0800 h (before the meal) and at hourly intervals from then until 1600 h. Plasma glucose concentrations throughout the sampling period were slightly, but significantly (P < 0.01), greater in patients with IGT than in normal individuals. Patients with NIDDM had markedly elevated glycemic excursions, greater than either of the other two groups (P < 0.002). Both plasma immunoreactive insulin and C-peptide concentrations from 0800-1600 h were higher (P < 0.002-0.001) in patients with either IGT or NIDDM than in the group with normal glucose tolerance. Although day-long plasma immunoreactive insulin and C peptide concentrations were higher, on the average, in patients with IGT compared to those with NIDDM, the difference was not statistically significant. Plasma proinsulin concentrations were highest in patients with NIDDM (P < 0.002), lower in those with normal glucose tolerance (P < 0.002), and intermediate in patients with IGT. When the calculated "true" insulin concentration was determined by taking the proinsulin content into consideration, patients with IGT had the highest day-long levels, with the lowest values found in the control population (P < 0.002). Although absolute values varied as a function of obesity, the generalizations outlined above were found in both weight groups. These results show that ambient plasma proinsulin concentrations increase as glucose tolerance declines. However, true plasma insulin concentrations in response to mixed meals remain highest in patients with IGT, lowest in normal individuals, and intermediate in patients with NIDDM. Thus, previous conclusions that absolute day long plasma insulin concentrations are not lower than normal in patients with NIDDM do not appear to result from an inability to differentiate true insulin from proinsulin. PMID- 8421102 TI - Growth hormone secretion by human peripheral blood mononuclear cells detected by an enzyme-linked immunoplaque assay. AB - In order to evaluate the secretion of GH from human peripheral blood mononuclear cells (PBMC) and its possible role as a modulator of lymphoproliferation, we have developed a hormonal enzyme-linked immunoplaque assay. This assay captures GH between a monoclonal and polyclonal antibody. This is followed by adding substrate and a horseradish peroxidase-conjugated antibody against the polyclonal antibody which produces violet colored plaques where GH has been secreted. This assay is sensitive, specific, highly reproducible, and can detect picogram quantities of GH. Using this assay we have detected GH secretion from approximately 1% of human PBMC under unstimulated conditions. Regression analysis showed a linear relationship between the number of cells plated and the number of GH plaques formed. Therefore, GH plaques were formed by single cells or its progeny and did not represent aggregation of secreting cells. Preincubation of PBMC with cycloheximide, a protein synthesis inhibitor, completely abolished the formation of GH plaques which suggests that the PBMC were responsible for the synthesis of the secreted GH. In addition, we have also observed that stimulation of human PBMC with T-cell mitogens, Concanavalin A and PHA and the cytokine IL-2 led to significant increases in GH plaque area and number whereas stimulation with LPS, a B cell mitogen, was ineffective. The PHA and IL-2 induced increase in plaque number suggests that they can recruit noncommitted lymphocytes to actively secrete GH which raises the possibility that this secreted GH may serve as a growth factor in T cell proliferation. We conclude that this immunoplaque assay may be useful in evaluating the secretion of other peptides from human immune cells. PMID- 8421103 TI - Amino acid substitutions in the intracellular part of the growth hormone receptor in a patient with the Laron syndrome. AB - By complementary DNA cloning we have identified two amino acid substitutions in the intracellular region of the human GH receptor in a child with growth failure and clinical features of the Laron syndrome. At the second position of codon 422 a G to T transversion changes a cysteine residue to phenylalanine, whereas at the first nucleotide of codon 561 an alteration from C to A leads to the substitution of threonine for proline. Direct analysis of exon 10 of the GH receptor gene showed that both nucleotide substitutions reside on the same chromosome and were inherited from the patient's mother. Evaluation of DNA from 10 additional prospectively recruited children with growth failure and a clinical picture similar to the index case did not reveal any nucleotide alterations in codons 422, 560, or 561. These observations represent the first demonstration of variation within the intracytoplasmic part of the human GH receptor and indicate that mutations occurring at multiple locations within the receptor gene may lead to the same clinical phenotype. PMID- 8421104 TI - Evidence for abnormal regulation of insulin receptors in Friedreich's ataxia. AB - Friedreich's ataxia is associated with a high incidence of diabetes mellitus. We have previously demonstrated that insulin resistance is present in nondiabetic patients with Friedreich's ataxia. This was associated with a reduction in the affinity of insulin receptors on freshly isolated monocytes. In this study we investigated the ability of the monocyte insulin receptor to acutely alter its affinity in response to oral glucose. Glucose and insulin concentrations were higher in the patients with Friedreich's ataxia after an oral glucose load, consistent with the presence of insulin resistance. The normal increase in the affinity of insulin receptors on monocytes 5 h after oral glucose was absent in the five patients with Friedreich's ataxia. Receptor affinity actually decreased in three of the five patients. These findings support the concept that a membrane abnormality that alters the binding function of the insulin receptor is present in these patients. PMID- 8421105 TI - The relative expression of mutant and normal thyroid hormone receptor genes in patients with generalized resistance to thyroid hormone determined by estimation of their specific messenger ribonucleic acid products. AB - Generalized resistance to thyroid hormone (GRTH), is a syndrome of reduced tissue responsiveness to thyroid hormone. So far, mutations linked to GRTH have been only detected in the hormone-binding domain of the human thyroid hormone receptor (hTR)-beta gene. Although there is no doubt that these mutations result in abnormal hTRs, there is a conspicuous lack of correlation between the severity of clinical manifestations and the degree of functional impairment of the mutant hTRs. In this work we examined whether variable expression of mutant genes relative to the normal genes could explain the observed discrepancies. The relative amounts of mutant and normal hTR beta and normal hTR alpha messenger RNAs in fibroblasts from normal subjects and those from individuals with GRTH were estimated by coamplification of their complementary DNA products. Heterozygous subjects with GRTH from two families manifesting differences in the severity of clinical manifestations expressed equally both normal and mutant hTR beta alleles. Furthermore, there was no compensatory increase in the expression of the normal hTR alpha gene in these individuals nor in fibroblasts from members of a third family with homozygous deletion of the hTR beta gene. In vitro treatment with thyroid hormone did not affect the results. It is concluded that the apparent discrepancies between the functional impairment of the mutant hTRs and the clinical manifestations of GRTH are not due to quantitative differences in the expression of the normal or mutant hTR genes but more likely to variations in the interactions of the mutant hTRs with the normal hTR beta, hTR alpha and nuclear stabilization factors. PMID- 8421106 TI - Increased in vitro thyrotropic activity of partially sialated human chorionic gonadotropin extracted from hydatidiform moles of patients with hyperthyroidism. AB - The intrinsic thyrotropic activity of hCG purified from normal pregnancy urine has been demonstrated in several laboratories. hCG has a specific thyrotropic potency of about 0.04-0.5 microU bovine (b) TSH/IU hCG, depending on the bioassay system. The corresponding potency for hCG derived from pathological sources, such as hydatidiform moles and choriocarcinoma tissue, or from the serum of these patients has not been studied as extensively. Since the biological activity of glycoproteins can be strongly influenced by variations in the oligosaccharide side-chain composition, we have investigated the effect of anion exchange chromatography on the thyrotropic potency of hCG derived from the hydatidiform mole and serum of three hyperthyroid patients with molar pregnancy. The activity for the fraction of total molar hCG immunoreactivity that was not retained by an anion exchange column (0.18-0.90 microM bTSH/IU hCG) was about twice that of the corresponding serum and molar hCG fraction eluting during the NaCl gradient elution (0.08-0.40 microU bTSH/IU hCG). The unretained hCG fraction corresponds to a previously described hCG precursor that is partially desialated in the C terminal region of the beta-subunit. PMID- 8421107 TI - Abnormal facilitative glucose transporter gene expression in human islet cell tumors. AB - Our previous studies have shown that increased expression of GLUT1/erythrocyte and GLUT3/brain type glucose transporter genes in human tumors is associated with cellular transformation. We have now determined the levels of messenger RNAs (mRNAs) encoding these two glucose transporter isoforms as well as that of GLUT2/liver isoform in insulin-, glucagon-, and gastrin-secreting islet cell tumors. Northern blot analysis and reverse transcriptase-polymerase chain reaction revealed the presence of GLUT1 and GLUT3 mRNA in all human islet cell tumors and normal islets examined. In contrast, GLUT2 mRNA, which is present at high levels in normal islets, was not detected in insulinomas or other types of islet cell tumors. These results imply that GLUT1 and GLUT3 are primarily responsible for glucose uptake by these tumors. PMID- 8421108 TI - Alterations in the kinetics of C-peptide and insulin secretion in hyperthyroidism. AB - Previous studies investigating the mechanisms underlying the hyperinsulinemia observed in hyperthyroid subjects have demonstrated increased, normal, or reduced insulin secretory rates when peripheral concentrations of C-peptide were used as a marker of beta-cell function. In this study, using individually derived C peptide kinetic parameters, insulin secretion rates were calculated directly from plasma C-peptide concentrations in 13 hyperthyroid and 13 euthyroid control subjects matched for age, weight, and sex. Eight subjects in each group were studied during a 24-h period in which they ate three mixed meals, whereas the remaining five were studied during a 3-h hyperglycemic clamp. Although insulin secretory rates under basal conditions in both groups were similar, the hyperthyroid group had an enhanced insulin secretory response to meals and, accordingly, the total amount of insulin secreted over 24 h was significantly greater (P < 0.02) in this group. Insulin secretory rates were also 50% higher in the hyperthyroid subjects during the hyperglycemic clamp at a time when glucose levels in both groups were comparable. Despite these differences in secretion, the C-peptide concentrations were not significantly different. Analysis of C peptide clearance kinetics using multivariate analysis demonstrated that the mean clearance rate of C-peptide was significantly increased (P < 0.02) in the hyperthyroid group. Thus, stimulated insulin secretion rates are significantly increased in thyrotoxicosis possibly reflecting an increased sensitivity of the beta-cell to glucose in subjects who are hyperthyroid. However, due to the rapid clearance of C-peptide from the circulation in the setting of hyperthyroidism, differences in beta-cell secretory responses between hyperthyroid and euthyroid subjects may not be evident by measurement of C-peptide levels alone. PMID- 8421109 TI - Lack of intrathyroidal tumor necrosis factor alpha in Graves' disease. AB - In vitro tumor necrosis factor alpha (TNF alpha) exerts a synergistic action on HLA class II expression and a cytotoxic action in FRTL-5 cells. Therefore, a role for TNF alpha as a local mediator of cell destruction in thyroid autoimmunity has been postulated. To elucidate the in vivo significance of these and other in vitro findings for the pathophysiology of Graves' disease we investigated 11 thyroid glands of patients suffering from Graves' disease for TNF alpha. In situ hybridization was done with a TNF alpha probe synthesized with T7 polymerase on a 750-base pair EcoRI fragment of the coding region. Primers at positions 152 and 854 in the TNF alpha copy DNA sequence were used for reverse polymerase chain reaction (PCR) amplification of TNF alpha in 5 patients. Immunohistological staining for TNF alpha was done with a mouse monoclonal antibody. We could not detect any TNF alpha and TNF alpha messenger RNA in thyroid tissue of 11 patients suffering mostly from relapsing Graves' disease by immunohistology and in situ hybridization as well as reverse PCR, respectively. A faint signal could be detected by reverse PCR in control thyroid tissue from a patient with recurrent goiter. This lack of intrathyroidal TNF alpha in relapsing Graves' disease is in accordance with a lack of increased TNF alpha production by T cell clones isolated from Graves' disease thyroid glands and contrasts with previous in vitro results. Since protective TNF actions have been demonstrated in other autoimmune diseases it could therefore be envisaged that the lack of intrathyroidal TNF alpha may be associated with the relapse of Graves' disease in our patients. PMID- 8421110 TI - Successive removal of periodontal tissues. Marginal healing without plaque control. AB - The aim of the present study was to compare periodontal healing after successive removal of periodontal tissue components, from the alveolar bone to the dentin surface. The prevailing tissue reaction when adhering PDM was left on the exposed roots was that most of the bone tissue that had been removed from the buccal surfaces had regenerated and the integrity of the PDM between the new alveolar bone and cementum surface had been reestablished. On exposed etched cementum surfaces, 2 prevailing healing results were recorded. In half the number of the roots, the root surfaces were covered by connective tissue with fibers running parallel to the root surfaces in a capsule-like arrangement. The other prevailing reaction was a thin epithelial cell-lining running parallel to the root surfaces in close contact or partly penetrating the adjacent connective tissue. On exposed denuded dentin surfaces, gingival retraction was a constant finding, associated with an epithelial cell-lining of varying thickness sometimes with rete pegs and cyst-like formations. Gingival retraction was also a constant finding on etched dentin surfaces. This was associated with pathological pockets outlined by epithelial cell-layers of varying thicknesses. The significance of these findings were discussed with special emphasis on dynamics of recurrent periodontitis. PMID- 8421111 TI - Evaluation of a mouthrinse containing chlorhexidine and fluoride as an adjunct to oral hygiene. AB - Chlorhexidine and fluoride have valuable preventive roles in dental and oral diseases. There is also evidence that in caries prevention, together, they provide additive benefits. However, combined chlorhexidine/fluoride formulations have rarely been evaluated. The aim of this study was to determine whether a 0.12% chlorhexidine, 100 ppm F- mouthrinse provided adjunctive oral hygiene benefits compared to a minus active control rinse. The study was a randomised double-blind parallel design involving 102 subjects of whom 99 completed the 6 week experimental period. Subjects rinsed 2x per day for 1 min with 15 ml of the allocated rinse. Normal toothbrushing procedures were maintained throughout the study. At 6 weeks, plaque and gingivitis scores were significantly lower and the incremental reduction significantly larger in the active rinse group. However, as expected, toothstaining was significantly increased with the active rinse. It is apparent that this chlorhexidine/fluoride rinse could be used in those regimens recommended for other chlorhexidine formulations. The value of the formulation in caries prevention would seem worthy of further investigation. PMID- 8421112 TI - Lactoferrin in the gingival crevice as a marker of polymorphonuclear leucocytes in periodontal diseases. AB - This study examined lactoferrin (LF) levels in gingival crevicular fluid (GCF) and set out to test the hypothesis that LF could act as a marker of crevicular polymorphonuclear leucocytes (PMN). Therefore, 2 experiments were conducted: (a) to quantify total LF (ng/30 s sample) in GCF; (b) to correlate LF levels (ng/microliters) and PMN numbers (PMNs/microliters) in gingival crevicular washings (GCW). GCF was collected from 71 sites in a total of 22 patients. These sites were classified on the basis of clinical indices of gingivitis (GI) and pocket depth (PD) into three clinical groups: 'healthy', 'gingivitis' and 'periodontitis'. GCWs were obtained from an additional 63 sites in 21 patients. LF in GCF and GCWs was assayed by a sandwich ELISA. Total leucocyte and differential counts were performed on the GCWs. GCF LF (ng/30 s) correlated positively with GI (r = 0.418, p < 0.001), PD (r = 0.415, p < 0.001) and GCF volume (r = 0.624, p < 0.001). Gingivitis (n = 21) and periodontitis sites (n = 24) demonstrated significantly higher (p < 0.05) total GCF LF than healthy (n = 26) sites. In GCWs LF (ng/microliters) showed stronger correlations with clinical indices (GI: r = 0.452, PD: r = 0.513, p < 0.001) than did PMN numbers (PMNs/microliters) (GI: r = 0.279, PD: r = 0.388, p < 0.05). LF correlated strongly with PMNs in GCWs (r = 0.531, p < 0.001) and provides a simple and effective marker of crevicular PMN numbers. PMID- 8421113 TI - Marginal bone loss in the primary dentition of patients with juvenile periodontitis. AB - 118 patients with juvenile periodontitis (JP), diagnosed when the patients were 13-19 years old, were studied retrospectively with respect to radiographic marginal bone loss in the primary dentition, experienced when the patients were 5 12 years old. 168 other 13-19 year old patients without any signs of bone loss in the permanent teeth were used as a reference group. The JP patients were classified into 2 groups according to the number of sites with bone loss in the permanent dentition: JP group I having 1 site with bone loss (n = 45) and JP group II having > or = 2 sites with bone loss (n = 73). It was found that 35 patients (52%) of JP group II displayed 1 or more sites with bone loss in the primary dentition during the age of 5-12 years. The corresponding numbers for JP group I and the reference group were 9 (20%) and 8 (5%) respectively. These findings indicate that juvenile periodontitis, at least in some individuals, may have its onset already in the primary dentition. PMID- 8421114 TI - The prevalence and severity of cyclosporin and nifedipine-induced gingival overgrowth. AB - The gingival health of 32 renal transplant patients who were medicated with cyclosporin was compared with a similar cohort of 23 renal transplant patients medicated with both cyclosporin and nifedipine. Both groups of patients had been taking the above medication for at least 3 months. Plaque scores, gingival inflammation and probing depths were similar for both groups. Patients medicated with the combination of nifedipine and cyclosporin had a significantly higher gingival overgrowth score (p = 0.046) when compared with the group receiving cyclosporin alone. The incidence of clinically significant overgrowth (i.e., overgrowth > 30% which would require surgical intervention) was similar in both groups. Gingival overgrowth was not related to cyclosporin dosage. It is concluded that patients taking cyclosporin or cyclosporin and nifedipine experience gingival overgrowth and that the severity of the overgrowth is greater in patients taking the combined therapy. The levels of plaque and gingival inflammation appear to be associated with this phenomenon. PMID- 8421115 TI - Relationship between periodontal probing velocity and gingival inflammation in human subjects. AB - Little is known about the biophysical characteristics of the dentogingival junction in response to the development or resolution of inflammation. The Toronto Automated Periodontal Probe (TAPP) provides an estimate of the integrity of the dentogingival junction by measuring intrapocket probing velocity. The aim of this study was to measure changes of probing velocity in inflamed human periodontium before and after subgingival debridement. 32 subjects exhibiting gingival inflammation were selected; 29 completed the study. Gingival index (GI), plaque index (PLI), bleeding index (BI) and the rate of gingival crevicular fluid flow (CFF) were measured as concomitant variables. The experimental group (N = 16) received scaling, root planing and oral hygiene instruction at baseline. The control group (N = 13) received no treatment until after 28 days. Subjects were seen at baseline, day 14, 21 and 28 for measurement of probing velocity and concomitant variables on 6 index teeth. At day 28, the control group was treated and then reassessed 28 days later. The experimental group showed a reduction of 51.6% for mean crevicular fluid flow (p < 0.0001), 79.7% for mean plaque index (p < 0.0001), 58.0% for mean gingival index (p < 0.0001), and 72.0% for mean bleeding index (p < 0.01) at day 28, confirming that inflammation was reduced compared with baseline. No significant changes were observed in the control group until after treatment. The velocity of probing and the formation of a plateau in the velocity profile were recorded.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421116 TI - The effect of a mouthrinse containing chlorhexidine and fluoride on plaque and gingival bleeding. AB - The aim of this study was to test the effect of a rinse with 0.05% sodium fluoride and 0.05% chlorhexidine on plaque and gingival inflammation compared with a placebo without these agents. In a double-blind study, 47 adults with > 20 teeth and a CPITN score > 1 but < 4 were randomised into test and control groups. After baseline assessments for plaque, bleeding and stain, teeth were professionally cleaned. Subjects were asked to rinse for 30 s with 10 ml of the respective test or placebo rinse after normal oral hygiene for 8 weeks. 39 subjects completed the study. There was no significant difference in the 2 groups at baseline with respect to either plaque or bleeding scores. After scaling and 8 weeks use of the test rinse, there were significant reductions (p < 0.001) in both plaque and bleeding. The control group showed no significant reduction in plaque scores after 8 weeks, but a significant (p < 0.05) reduction in bleeding. However, this reduction was significantly greater (p < 0.001) in the test group than in the control. The test group had a significantly greater (p < 0.05) stain score than the control at baseline. After scaling and rinsing for 8 weeks, stain scores were lower for both groups compared to baseline but reached significance (p < 0.05) only for the control group. It is concluded that, as an adjunct to normal oral hygiene, the chlorhexidine/fluoride rinse had a significant inhibitory effect on plaque and bleeding but its effect on staining is uncertain. PMID- 8421117 TI - Evaluation of reliability and reproducibility of dental indices. AB - Evaluation of periodontal therapy involves the use of several oral indices to describe the health status of hard and soft tissues. It was the objective of the present study to evaluate the reliability and reproducibility of some of these indices. A calibration and standardization session was designed to calibrate 10 examiners and a "gold standard" (an experienced examiner) in evaluating the following indices: the Volpe-Manhold calculus index (VM), the Lobene stain index (SI), a non-invasive modification of the Loe-Silness gingival index (GI), the papillary bleeding score (PBS) of Loesche, and the plaque index (PI) of Quigley Hein as modified by Turesky. For each index, the average intraclass correlation was calculated between two subject visits. The highest intraclass correlation, 0.94, was found for PBS. The intraclass correlation for PI was 0.70 and for VM, 0.65. The lowest intraclass correlations were for stain, 0.47, and GI, 0.25. Intra-subject correlations between the 2 visits were good for all indices, but were best for PBS, followed by VM. PBS seems to be the most reliable index (both intra- and inter-examiner) for measuring the oral health status and is therefore recommended for use in clinical studies. PMID- 8421118 TI - The effects of a chlorhexidine toothpaste on the development of plaque, gingivitis and tooth staining. AB - Chlorhexidine has found many uses in clinical dentistry as an antiplaque agent. To date, effective chlorhexidine-containing toothpastes have not been made available. This study was the first phase in the evaluation of a 1% chlorhexidine toothpaste, formulated to ensure a high availability of the contained antiseptic. The study was a 19-day, randomised double-blind placebo-controlled, crossover experiment al gingivitis clinical trial employing 14 healthy human volunteers. From a zero plaque and low gingivitis baseline, the development of plaque, gingivitis and dental staining was measured on days 12 and 19 of the 2 study periods. The toothpaste was used 2x a day as a slurry rinse with no other form of oral hygiene. The washout period was 21 days. The development of plaque and gingivitis was statistically and clinically highly significantly reduced during the use of the active compared to the placebo product. Conversely, dental staining was statistically and clinically significantly increased. The product deserves further evaluation for potential clinical use. PMID- 8421119 TI - Calculus removal from multirooted teeth with and without surgical access. (I). Efficacy on external and furcation surfaces in relation to probing depth. AB - The purpose of this study was to evaluate the efficacy of the calculus removal from multirooted teeth after closed root planing, open root planing and use of a rotary diamond for the furcation area. The effect of pocket depth on the effectiveness of calculus removal was also examined. 30 first and second lower molars scheduled for extraction, with a calculus index > or = 2 and a degree II or III furcation involvement, were divided into 3 groups: 10 molars were scaled and root planed using a closed approach; 10 molars were scaled and root planed using an open approach; 10 molars were scaled and root planed with an open approach and rotary diamond was used for removal of deposits in the furcation area. After extraction, the teeth were assessed in a stereomicroscope and the % of residual calculus was calculated on external and furcation surfaces. The % of residual calculus on the external surfaces was significantly higher after closed than open root planing (p = 0.002). Pocket depth affected the effectiveness of scaling and root planing, with more residual calculus observed for depths > or = 7 mm for both groups. Differences between the 3 groups in the % of residual calculus on furcation surfaces were statistically significant (p < 0.0001 and p < 0.0005). The most effective method was the combination of open root planing and rotary diamond. More calculus was observed in all groups for pocket depths > or = 7 mm but the difference was significant only in the closed group (p = 0.006).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421120 TI - Periodontal disease and dental caries in relation to primary IgG subclass and other humoral immunodeficiencies. AB - 22 females and 3 males with primary hypogammaglobulinaemia (n = 6) or IgG subclass deficiencies with (n = 3) or without (n = 16) concomitant IgA deficiency were examined for periodontal disease and dental caries. Only 1 patient showed more tooth loss than that found in the normal Swedish population. 1 patient demonstrated advanced periodontal disease. No patient exhibited more severe dental caries than that of comparable normal Swedes. Microbiological samples from periodontal pockets and saliva showed recovery of potential periodontopathic and cariogenic bacteria within normal ranges. This study could not support the notion that immunodeficient subjects exhibit an increased risk of developing periodontal disease or dental caries. PMID- 8421121 TI - Teaching site: the waiting room. PMID- 8421122 TI - Role stress and strain among nondoctorally prepared undergraduate faculty in a school of nursing with a doctoral program. AB - Initiation of a doctoral program within existing schools of nursing causes significant change in organizational structure and function. The role expectations of the current faculty change as well. The purpose of the qualitative study was to identify if role stress and strain are present in nondoctorally prepared undergraduate nursing faculty in a southern university school of nursing with a doctoral program. The design was a qualitative, open ended interview. The primary research question was: Does the initiation of a doctoral program in a school of nursing impose role stress and strain on nondoctorally prepared faculty members? Academic educators will find our results interesting because faculty report that role stress and strain affect both their teaching and decisions to remain in academia. Use of these findings may contribute toward improved role relationships, better role performance, greater job satisfaction, and decreased job turnover. PMID- 8421123 TI - Postdoctoral study in a multidisciplinary research center: an alternative to more traditional nurse fellowships. AB - This article examines postdoctoral study in a multidisciplinary research setting. The author describes the offerings of one particular multidisciplinary center, the Monell Chemical Senses Center, and highlights the advantages and disadvantages of a fellowship in such a setting. The relevance of this option for nursing is discussed. PMID- 8421124 TI - The "real world" of hospital nursing practice as perceived by nursing undergraduates. AB - The purpose of this study was to examine the expectations of senior nursing undergraduates in the "real world" of hospital nursing practice, with particular regard to the ethical dimensions of their role. The method was qualitative; specifically, a grounded theory approach was used. The sample consisted of 23 senior baccalaureate nursing students, from a possible population of 120, who were in their final clinical rotation before graduation. Informants were volunteers who gave informed consent, having been briefed on the purposes of the study and how their confidentiality would be protected. Data were collected through audiotaped interviews and clinical logs and analyzed through the constant comparative method. The study concluded that (1) senior nursing students were not naive about the reality of practice; (2) most perceived themselves as fairly powerless; (3) they voiced a commitment to the ethical principle of respect for the client as evidenced by listening, providing information, accepting values, and encouraging a climate for self-determination; (4) they experienced guilt when they did not "say something"; and (5) they expressed disappointment that nurses did not "stand up" for patients. PMID- 8421125 TI - Inquiry, insights, and history. A new phase. PMID- 8421126 TI - Learning style research: a critical review of the literature and implications for nursing education. AB - Much has been written about the implications of learning styles on the teaching/learning process. As a result, several articles related to learning styles can be found in the nursing literature. Although learning styles is one factor that should be considered when planning teaching strategies, its impact may be overrated. Many studies have ignored the complexity of the learning process by attempting to show a cause-and-effect relationship between cognitive/learning styles and achievement. Other factors that should be considered when developing teaching plans are discussed. PMID- 8421127 TI - Education. Downsizing--keep the focus on your mission. PMID- 8421128 TI - Effects of a participative intervention on high school students' image and valuation of nursing. AB - In this pretest, posttest, control group study, high school biology students (N = 451) were actively engaged in a dialogue with an investigator/change agent regarding specific nursing situations in which the image dimensions of caring, useful, scientifically knowledgeable, and autonomous were highlighted. The effects of this involvement and participation on students' image and valuation of nursing were assessed. The process was found to be highly effective in expanding students' image of nursing and moderately effective in increasing their valuation of nursing. PMID- 8421129 TI - International affairs. International exchange--students and faculty. PMID- 8421130 TI - Pay inequity: it's still with us. AB - This article explores reasons for the continued existence of pay inequity. Throughout the past 3 decades, pay equity proponents have applied a number of remedies for inequity. For example, civil rights legislation has helped somewhat, but narrow interpretations hinder the usefulness of law. Labor market surveys do more to perpetuate inequity than to cure it. Collective bargaining offered by traditional unions benefits nursing to a lesser extent than nurses commonly believe. Job evaluation systems combine with multiple pay scales and inequitable pay formulas to perpetuate discrimination. Finally, there remains a danger that nurses' own lack of sophistication regarding pay equity issues may work against the interests of the profession on the pay equity scene. Nurses, therefore, need to keep abreast of current pay equity issues. PMID- 8421131 TI - Therapeutic syncretism: a conceptual framework of persistence and change for international nursing. AB - International nursing, as well as the entire nursing profession, has committed itself to a change paradigm. An alternative to the change paradigm has been proposed to account for elements of stability and persistence. This alternative, therapeutic syncretism, is defined and described, emphasizing an individual's effect on the syncretic process. Finally, the Kingdom of Swaziland illustrates therapeutic syncretism in action as the nation, nurses, and indigenous healers reflect qualities of persistence and change. PMID- 8421132 TI - Legal and ethical issues. Whither ethics in nursing curricula? PMID- 8421133 TI - Schools with nursing centers: a 5-year follow-up study. AB - The article reviews the growing body of literature that examines academic nursing centers in the aggregate including the first such study conducted by the primary author 5 years earlier. A follow-up study to this original work was conducted to determine current demographics and faculty practice policies of schools or colleges of nursing that operate nursing centers and to compare these findings with those of schools without nursing centers. A survey was sent to the deans or directors of the 462 National League of Nursing (NLN)-accredited baccalaureate nursing programs. Although there were 362 respondents to the survey (78 per cent), only 41 indicated they had a nursing center. Schools with nursing centers were significantly more likely to be in public institutions (P = .05), and offer master's degrees (P = .01). They also were significantly larger (P = .01), with a mean of 34 full time-equivalent (FTE) faculty members. Then the administrative policies of schools with nursing centers (N = 41) were compared with those of schools without centers but with practicing faculty (N = 187). Requiring practice was not a common policy in either group. While about one-third of both groups had practice plans, the majority were described as informal. More schools with nursing centers had other formalized practice arrangements than did schools without centers (P = .01). Faculty practice was more likely to be a criterion for promotion (P = .05) but not for tenure in schools with nursing centers. An inverse relationship (P = .05) was found between the amount of practice revenue generated and the presence of a nursing center.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421135 TI - 50th Anniversary of the American Academy of Allergy and Immunology. 49th Annual Meeting. Chicago, Illinois, March 12-17, 1993. Abstracts. PMID- 8421134 TI - 42nd Annual Scientific Sessions. Anaheim, California, March 14-18, 1993. Abstracts. PMID- 8421136 TI - Effects of enteric parasitoses and HIV infection upon small intestinal structure and function in patients with AIDS. AB - We compared, retrospectively, the effects of infection in jejunal mucosa with the protozoa cryptosporidia or microsporidia and with the human immunodeficiency virus (HIV) upon mucosal structure and absorptive function in 29 AIDS patients. The presence or absence of protozoal infection was confirmed by transmission electron microscopy. Villus blunting and crypt hyperplasia were seen mainly in the parasite-infected groups, although two patients without parasites also had shortened villi. Absorptive functions, including disaccharidase-specific activities and D-xylose absorption, closely paralleled the degree of small intestinal alteration. Evidence of HIV-infected cells in jejunal mucosa was examined by RNA in situ hybridization and by antigen-capture ELISA of mucosal homogenates. We found evidence of HIV in almost half the patients, which did not correlate with intestinal injury or diminished absorption. PMID- 8421137 TI - Subclinical celiac sprue. Increasing occurrence and clues to its diagnosis. AB - We have reviewed the clinical records of 226 consecutive adult patients with celiac sprue diagnosed in our department from 1972 to 1989. The study period has been divided into three subperiods of 6 years each (1972-1977, 1978-1983, 1984 1989). From the first to the third subperiod a significant increase in the number of new diagnoses and of the proportion of patients recognized by minor symptoms has been observed. These patients represent 50% of our series in the last 3 years of the study and 70% in 1989. Diagnosing subclinical forms of celiac sprue has significantly lowered the mean age at diagnosis and the female/male ratio. First degree relatives of celiac patients; subjects who had a gluten-free diet for a period during childhood; patients with short stature, anemia, or amenorrhea of no obvious cause; or patients with unexplained immunological abnormalities are the groups in which most patients with subclinical celiac sprue have been found and in which potential patients should be sought. Helpful diagnostic tools include antigliadin antibody testing and the observation of absent or reduced Kerckring folds in the descending duodenum in the course of upper gastrointestinal endoscopy. PMID- 8421138 TI - The gastroenterologist-patient relationship. AB - A strong doctor-patient relationship is a vital need, especially in gastroenterology, a specialty fraught with problems of ambience, logistics, invasiveness of both diagnostic and therapeutic techniques, and frequent inclusion in therapeutic trials. Studies designed to assess the degree of emotion aroused by endoscopic examination show the sensitivity of many patients to them and the emotion also generated at times in the endoscopist. In this era of rapid technological advancement, it is essential to pursue this problem at the cultural level. The solution lies both in greater maturity on the part of the patient and in the psychological as well as technological training of gastroenterologists so that they may regain the status of "complete" doctor. PMID- 8421139 TI - Malignant duodenocolic fistula. The role of extended surgery. AB - Malignant duodenocolic fistula is a seldom-seen complication of advanced right colon carcinoma. We describe four patients who illustrate the sometimes rewarding results of aggressive treatment. Because duodenocolic fistula is always associated with severe nutritional impairment, operation should be deferred until total parenteral nutrition (TPN) and blood transfusions have improved the clinical state. Whenever feasible, resection offers the best treatment; lesser techniques such as bypass and exclusion result only in minimal palliation. A direct approach and fistula disconnection are contraindicated. The benefit of exploration should almost always be offered, even in such secondary fistulas, as a better quality of life and long-term survival are realistic goals. PMID- 8421140 TI - Role of infectious agents in exacerbations of ulcerative colitis in India. A study of Clostridium difficile. AB - Fifty patients with idiopathic ulcerative colitis--25 with acute exacerbation of the disease (Group I) and 25 in quiescent phase (Group II)--were studied. None of the patients had a history of recent exposure to antimicrobial drugs or hospitalization. Evidence of infection with protozoal and bacterial agents and/or presence of Clostridium difficile toxin was demonstrated in eight (32%) patients in group I and one (4%) patient in group II (group I vs. group II, p < 0.05; chi 2 test with Yate's correction). In five of the six patients with demonstrable Clostridium difficile toxin in the stool, one patient with Entamoeba trophozoites, and another with Salmonella infection, the exacerbation responded clinically and endoscopically to specific antimicrobial therapy. Another two patients who had Entamoeba histolytica cysts in the stool had no change in their clinical status with metronidazole. We conclude that infectious agents are responsible for some of the exacerbations in patients of ulcerative colitis in a tropical country like India, where careful microbiologic examination is in order in every acute exacerbation of this disease. This contrasts with the findings in developed countries. PMID- 8421141 TI - Segmental ischemic colitis associated with nonsteroidal antiinflammatory drugs. AB - We have seen two cases of a segmental ischemic colitis develop during nonsteroidal antiinflammatory drug (NSAID) treatment. No other possible etiologic factors were shown. The short-term clinical course and the follow-up were uneventful. NSAIDs have been reported to cause different lesions in the large bowel, either by worsening preexisting colonic diseases or by inducing a primary pathology. We suggest that ischemia should be considered a possible mechanism of NSAID-associated colitis. Such ischemic colitis, not triggered by severe cardiovascular disease or operation, may be related to NSAIDs more often than currently recognized. PMID- 8421142 TI - Ischemic hepatitis in cirrhosis. Rare but lethal. AB - We report two cases of ischemic hepatitis in patients with alcoholic cirrhosis. In both, hepatic ischemia was induced by hemorrhagic shock and severe sepsis. Despite control of the bleeding and restoration of normal hemodynamics, liver failure deteriorated to hepatic coma and death in both cases. Ischemic hepatitis occurred in 1.5% of 130 consecutive cases of cirrhosis admitted for hemorrhage on our medical intensive care unit. Although cirrhotic patients run an increased risk of ischemic hepatitis, our experience and our review of the literature indicate that this condition is rare in these patients. PMID- 8421143 TI - Celiac sprue and diabetes mellitus. AB - The frequent association of celiac sprue (CS) and insulin-dependent diabetes mellitus (DM) which may be the result of interplay between genetic, hormonal, and immunologic factors has obvious therapeutic implications. A gluten-free diet may improve the diarrhea in some patients with DM where the reason for diarrhea is underlying celiac sprue; the diet may also improve control of DM by normalizing the serum hormonal profile. PMID- 8421144 TI - Chronic diffuse varioliform gastritis in a child. Total gastrectomy for acute massive bleeding. AB - Chronic diffuse varioliform gastritis is an uncommon, subacute, inflammatory gastric mucosal disease characterized by swollen congested rugae and disseminated mucosal erosions. The entity is exceptionally rare in children. The spectrum of reported symptoms is broad; frank hematemesis has never been reported in childhood. We present a child in whom the disease had a remarkably unusual clinical course. Because many caretakers were unaware of the existence of the disease in children, the patient had numerous hospitalizations and surgical procedures, until massive gastric bleeding resulted in unavoidable emergency total gastrectomy. Microscopical examination and immunofluorescent staining of the gastric mucosa confirmed the diagnosis of chronic diffuse varioliform gastritis. PMID- 8421145 TI - "Postgastrectomy" bezoar secondary to gastric cancer. AB - Phytobezoars are frequently associated with partial gastrectomy and are generally considered benign. However, we report two patients who developed a gastric bezoar due to malignancy at the anastomotic site. Development of intragastric bezoar may be an indication of neoplastic growth sufficient to obstruct a small gastric outlet. Consequently, we conclude that endoscopic or surgical evaluation of the anastomotic site is necessary in patients who develop bezoars after gastric surgery. PMID- 8421146 TI - The brown bowel syndrome and gastrointestinal adenocarcinoma. Two complications of vitamin E deficiency in celiac sprue and chronic pancreatitis? AB - The brown bowel syndrome is a rare disorder caused by vitamin E deficiency occurring in malabsorption syndromes. In patients with celiac sprue and chronic pancreatitis, the death rate from malignancy is high. We believe that vitamin E deficiency is responsible for the development of the brown bowel syndrome and may be partially responsible for the high incidence of malignancy in patients with celiac sprue and chronic pancreatitis. We report such a patient, and review the literature. PMID- 8421147 TI - Leptospirosis with acute acalculous cholecystitis and pancreatitis. AB - Severe Leptospira autumnalis infection was associated with acute acalculous cholecystitis and pancreatitis in a 66-year-old man. He was successfully treated with antimicrobial agents and supportive therapy, including hemodialysis. We review these uncommon manifestations and the effectiveness of antimicrobial therapy in advanced leptospirosis. PMID- 8421148 TI - Ulcerative colitis, primary biliary cirrhosis, and chronic pancreatitis: coincident or coexistent? AB - We report a 45-year-old woman with chronic ulcerative colitis complicated by primary biliary cirrhosis and concomitant chronic pancreatitis. The combination of primary biliary cirrhosis and ulcerative colitis has only rarely been reported, and the association of these diseases with chronic pancreatitis is even rarer. These systemic manifestations of ulcerative colitis should be looked for more often. PMID- 8421149 TI - Biliary and colonic cutaneous fistula successfully treated by endoscopic insertion of biliary stents. AB - We report a case in which a large, postoperative cutaneous biliary and a colonic fistula were successfully treated by means of endoscopic insertion of two endoprostheses into the common bile duct. Twenty-four hours after the procedure, bile flow through the leak ceased completely while the colonic fistula closed in 6 days. Six months after the treatment, when a new endoscopic cholangiography showed no evidence of leak or stricture, the stents were removed. For the treatment of postoperative biliary leaks, insertion of endoprostheses is a good and safe alternative to surgery. PMID- 8421150 TI - Helicobacter pylori and gastroduodenal disorders in India--lessons from epidemiology. AB - The relationship of Helicobacter pylori and gastroduodenal disorders should be compared in the developing (DG) and developed (DD) nations (Table 1) for a better understanding of the etiopathogenesis of these disorders. Although chronic gastritis is widespread in the DG nations, benign and malignant gastric ulcer (GU) are uncommon. Repeated reinfections and longer duration of H. pylori infection in the fundus-body mucosa of patients with duodenal ulcer from the DG nations explain the lower values of maximal acid output reported. The lower prevalence of gastric metaplasia in patients with duodenal ulcer (DU) in the DG nations compared to patients in the DD nations suggests that DU in different populations results from varying mechanisms. PMID- 8421151 TI - Fulminant herpes hepatitis mimicking hepatic abscesses. AB - Fulminant hepatitis due to herpes simplex virus (HSV) in adults is a rare and deadly disease. We describe a 23-year-old woman with a 20-year history of Crohn's disease (CD) who was hospitalized with an acute febrile illness and diarrhea. A computed tomography (CT) scan of the abdomen demonstrated an intramural sigmoid colon abscess and multiple abscesses in the liver. Despite high-dose parenteral corticosteroids and broad-spectrum antibiotics, the patient remained acutely ill, with high fever and markedly elevated serum transaminase levels, but no jaundice. Sigmoid resection and wedge liver biopsy were performed at laparotomy. Histologic examination documented HSV-type intranuclear inclusions and inflammation with necrosis in both the sigmoid colon and liver specimens. The patient subsequently died despite parenteral acyclovir treatment. Although rare, fulminant hepatitis due to HSV simplex virus should be considered in the differential diagnosis of all patients with severe hepatitis. Of special note, the necrotizing liver lesions may be mistaken for pyogenic abscesses on CT scan. PMID- 8421152 TI - Primary small noncleaved cell lymphoma of the liver. Report of an adult case in complete remission after treatment with combination chemotherapy. AB - Open liver biopsy in a 34-year-old woman with hepatosplenomegaly showed small noncleaved cell lymphoma. Except for an enlarged spleen, there was no evidence of other sites of involvement. She was treated with combination chemotherapy and is alive and free of disease > 5 years after diagnosis. We believe this to be the first reported case in an adult of primary hepatic or hepatosplenic lymphoma of the small cleaved cell type with long-term disease-free survival. PMID- 8421153 TI - Low frequency of endoscopic esophagitis in Asian patients. AB - The aim of this study was to determine the frequency of endoscopic esophagitis in patients seen for upper gastrointestinal complaints in an Asian center. We studied a consecutive series of 11,943 patients undergoing diagnostic esophagogastroduodenoscopy at our unit over a 10-year period. Three hundred and eighty-nine patients (3.3%) had endoscopic esophagitis with no other significant lesion (primary esophagitis), whereas 143 (1.2%) had esophagitis associated with peptic ulcer or gastric or duodenal malignancy (secondary esophagitis). In contrast, peptic ulcer was diagnosed in 2,787 patients (23.3%) and gastric carcinoma in 286 (2.4%). The reported frequency of endoscopic esophagitis among patients undergoing endoscopy in Western countries varied from 9 to 23%. Our data therefore show that endoscopic esophagitis is much less common in Singaporean patients. PMID- 8421154 TI - Diaphragm disease of the ascending colon. Association with sustained-release diclofenac. AB - We describe the clinicopathological features of six patients, two with rheumatoid arthritis and four with osteoarthritis, in whom intake of sustained-release diclofenac for one or more years was associated with ulceration and or stricture of the ascending colon. All were referred for further evaluation of anemia and changes in bowel habits. Three had chronic watery diarrhea, one suffered from progressive constipation and subsequently needed a right hemicolectomy because of complete intestinal obstruction. In five patients, colonoscopy revealed single to multiple semilunar ulcers, predominantly localized on the crest of the haustra of the ascending colon. In five of six cases the lumen was narrowed, from slight accentuation of the haustrum to almost pinhole-like concentric stenosis. All except one patient had multiple diaphragm-like strictures. The macroscopic and microscopic appearances closely resembled those of similar lesions previously described in the terminal ileum in patients treated with nonsteroidal anti inflammatory drugs. It appears that the slow-release form of a nonsteroidal anti inflammatory drug, such as sustained-release diclofenac, predisposes to manifestations of such lesions in the ascending colon. PMID- 8421155 TI - Endogenous mucosal prostacyclin levels in newly developed, untreated ulcerative colitis. PMID- 8421156 TI - More causes of abdominal wall pain. PMID- 8421157 TI - The 13C-urea breath test, Helicobacter pylori infection, and the operated stomach. PMID- 8421158 TI - Biliary obstruction due to spontaneous hemorrhage into benign hepatic cyst. PMID- 8421159 TI - Spontaneous rupture of abdominal-wall hernia in a noncirrhotic patient with ascites. PMID- 8421160 TI - Acute colonic obstruction caused by anastomotic jejunal loop two years after antrectomy with antecolic Billroth II reconstruction. PMID- 8421161 TI - Tuberculosis of the anal canal. PMID- 8421162 TI - Sinusitis and atopy in human immunodeficiency virus infection. AB - Sinusitis is increased in patients with human immunodeficiency virus (HIV) infection. To determine the underlying mechanism(s), 37 HIV-positive patients were evaluated. HIV-negative controls included 21 with rhinosinusitis, 32 with atopy, and 16 without sinusitis. Twenty-two HIV-positive patients (59%) had sinusitis; 14 of them had AIDS. There was a significant association between sinusitis severity and stage of HIV infection (P < .05). IgE levels were higher in the HIV-positive patients, increased with disease progression, and were strongly correlated with sinusitis severity (P < .01). Of HIV-positive patients, 72% exhibited more than two positive skin tests compared with 24% of HIV-negative rhinosinusitis patients and 12.5% of controls (P < .05). Sinusitis is common in HIV-positive patients, especially those with AIDS. HIV causes an allergic diathesis with increased IgE levels and allergic reactivity. There is a significant correlation between IgE levels and sinusitis severity, suggesting sinusitis is part of this acquired atopic state. PMID- 8421163 TI - Efficacy of low doses of the polyethylene glycol derivative of interleukin-2 in modulating the immune response of patients with human immunodeficiency virus type 1 infection. AB - Interleukin-2 (IL-2) is a key cytokine in cellular immunity. Human immunodeficiency virus type 1 (HIV-1)-infected individuals lack IL-2 because of low CD4+ T lymphocyte numbers. In an attempt to enhance cellular immunity, low dose recombinant human (rh) IL-2 at 10 micrograms or 180,000 units or its polyethylene glycol (PEG) derivative at 9 micrograms or 36,000 units was given by intracutaneous injection to 8 HIV-1-infected men for 30 days. Participants had no evidence of opportunistic infection and received concurrent zidovudine. IL-2 treatment was nontoxic and elicited a local cellular response resembling classic delayed-type hypersensitivity (DTH) with local interferon-gamma production, even in anergic patients. Systemic responses included enhanced DTH responses to recall antigens, improved in vitro proliferative responses to mitogen, and enhanced NK cell activity. Peripheral leukocyte phenotype and virus titers were unchanged. Long-term studies of low-dose IL-2 are warranted to determine whether immunoenhancing effects can be sustained and if they are associated with improved clinical course. PMID- 8421164 TI - Relationship of hepatitis B virus infection to human immunodeficiency virus type 1 infection. AB - The Multicenter AIDS Cohort Study (MACS) was designed to study the natural history of human immunodeficiency virus type 1 (HIV-1) infection, including the relationship between hepatitis B virus (HBV) and HIV-1 infection. In total, 4954 homosexual men were recruited from April 1984 through March 1985 and have been followed up thereafter every 6 months. Hepatitis B surface antigen and hepatitis B core antibody were tested for at the first visit by RIA or EIA; HIV-1 antibody testing was done at each visit by ELISA and confirmed by Western blot assay. The role of HBV infection in HIV-1 seroconversion was studied by stratification for sexual behavior and disease visit by visit. The adjusted risk ratio was 2.02 for hepatitis B surface antigen carriers and 2.14 for hepatitis-immune cases compared with hepatitis B-susceptible subjects. Similar results were obtained using a logistic regression model. After taking into account changes in sexual behavior and disease over time, the authors conclude that past HBV infection remains suspect as a cofactor or as a surrogate for other factors associated with HIV-1 seroconversion. PMID- 8421165 TI - Trivalent attenuated cold-adapted influenza virus vaccine: reduced viral shedding and serum antibody responses in susceptible adults. AB - Trivalent cold-adapted recombinant (CR) influenza virus vaccines containing types A (H1N1 and H3N2) and B viruses were evaluated in two double-blind, placebo controlled trials. Susceptible adults were randomly assigned to receive the following vaccines by intranasal drops 1 month apart: two doses of trivalent vaccine, bivalent CR influenza A (Bi A) vaccine followed by monovalent B (Mono B) vaccine or vice versa, or two doses of placebo. All vaccines were well tolerated. Shedding of each of the three vaccine viruses was reduced after the first dose of trivalent vaccine compared with primary vaccination with Bi A or Mono B. Shedding was also reduced after second vaccinations, whether homologous (trivalent trivalent) or heterologous (Bi A/Mono B or Mono B/Bi A). Reduced viral shedding was associated with reduced serum antibody responses. Thus, both simultaneous and sequential inoculations of susceptible adults with CR influenza vaccine viruses result in reduced viral shedding and serum antibody responses. PMID- 8421166 TI - Adhesion of Staphylococcus aureus to surface-bound platelets: role of fibrinogen/fibrin and platelet integrins. AB - Platelets adhering to artificial or biologic surfaces have been implicated in the pathogenesis of catheter infections or endocarditis; however, the ligands involved in Staphylococcus aureus interaction with adherent platelets remain incompletely understood. Radiolabeled S. aureus Cowan I were incubated with purified platelets adherent to polymethylmethacrylate (PMMA) coverslips and washed, and adhesion was determined. Platelets promoted adhesion of S. aureus approximately 30-fold compared with adhesion to albumin-PMMA. In the presence of both plasma (1% vol/vol) and platelets, adhesion was extensively promoted, with 30% (of inoculated) S. aureus adherent (150-fold increase). Platelet pretreatment with anti-GPIIb/IIIa monoclonal antibodies or inhibitors of platelet activation decreased plasma-enhanced adhesion, suggesting a role of platelet activation in S. aureus adhesion. Plasma-enhanced adhesion was sensitive to thrombin antagonists, proteinase inhibitors, heparin, or antifibrinogen antibodies, indicating that fibrinogen/fibrin is necessary for bridging between adherent platelets and S. aureus. In conclusion, S. aureus adhesion to immobilized platelets may play a role in the pathogenesis of invasive bloodstream infections or endocarditis. PMID- 8421167 TI - Staphylococcus epidermidis adhesion to hydrophobic biomedical polymer is mediated by platelets. AB - A quantitative investigation on the effects of plasma proteins and platelets on the adhesion of Staphylococcus epidermidis RP62A to a hydrophobic biomedical polymer (National Heart, Lung, and Blood Institute reference polyethylene) was carried out under well-defined shear conditions approximating human blood circulation by using a rotating disk system. The results showed that contact activated platelets mediated S. epidermidis adhesion to the polymer surface. In the range of physiologic shear conditions, the adhesive coefficient (ratio of bacteria per unit area to the product of bacterial flux and the duration of the experiment) to platelets was significantly greater than to the protein-adsorbed polyethylene surface by at least one order of magnitude (P < or = .01). The presence of absorbed plasma proteins on polyethylene reduced the adhesion of S. epidermidis compared with that seen with the bare polymer surface. These studies show that S. epidermidis adhesion to polyethylene is mediated by contact activated platelets, not absorbed plasma proteins. PMID- 8421168 TI - Pneumococcal capsular polysaccharide-meningococcal outer membrane protein complex conjugate vaccines: immunogenicity and efficacy in experimental pneumococcal otitis media. AB - Vaccines composed of pneumococcal capsular polysaccharides (PS) conjugated to outer membrane protein complex (OMPC) from Neisseria meningitides group B bacteria were tested in the chinchilla otitis media model. Monovalent (types 6B and 23F), bivalent (6B+23F), and tetravalent (6B+14+19F+23F) PS-OMPC conjugate vaccines elicited significant total serum antibody responses against all four PS. Type 6B vaccine elicited IgG, IgM, and IgA antibodies after a single dose and an anamnestic IgG response after a second vaccine dose on day 28. Type 6B and 19F vaccines prevented or greatly attenuated pneumococcal otitis media after direct middle ear challenge with the immunizing serotype, type 14 vaccine was not protective by this challenge route, and type 23F pneumococci were not sufficiently virulent in chinchillas to test vaccine effectiveness. The promising results with two serotypes suggest the PS-OMPC conjugates may be useful in human infants. PMID- 8421169 TI - Genes involved in Haemophilus influenzae type b capsule expression are frequently amplified. AB - The genes involved in Haemophilus influenzae type b capsule expression are present as a duplication of an approximately 18-kb DNA segment (the Cap b locus). It has been shown previously that recombination occurs between the two copies of the repeat, resulting in deletion of one copy and loss of capsule expression at frequencies of 0.1%-0.5%. The present study tested the hypothesis that the duplicated arrangement could serve as a template for further amplification of capsule gene sequences. Southern hybridization analysis of 66 type b invasive isolates showed that amplifications exist and are moderately common (23/66 were amplified). In addition to three copies of the 18-kb repeat, four copies were detected in some strains, and up to five copies in 1 isolate. By ELISA, a five copy strain made about six times more capsular polysaccharide than did an isogenic two-copy derivative. The evolutionary significance of the duplicated arrangement may be its ability to rapidly amplify under conditions where it is advantageous to produce more capsule. PMID- 8421170 TI - Anti-capsular polysaccharide antibodies reduce nasopharyngeal colonization by Haemophilus influenzae type b in infant rats. AB - Haemophilus influenzae type b (Hib) conjugate vaccines reduced oropharyngeal carriage of Hib among children in Finland and the United States. To study the mechanisms of this reduction, a colonization model in infant rats with passively administered antibodies was developed. Of the pups, 94% were colonized 48 h and 64% 7 days after intranasal inoculation with approximately 2500 cfu of Hib. Intranasally administered anti-Hib antibodies, including human IgG and both serum and secretory IgA (sIgA) as well as murine monoclonal anti-Hib capsular polysaccharide (PS) of IgG1 isotype, given simultaneously with, before, or after bacteria, significantly reduced nasopharyngeal colonization by Hib. Hib colonization was also significantly reduced when the antibodies were given intraperitoneally, with a resultant anti-Hib PS serum concentration of > or = 7 micrograms/mL. Thus, anti-Hib PS antibodies, both sIgA and IgG, can function on the mucosal surface and prevent colonization. PMID- 8421171 TI - Pathogenesis of Campylobacter fetus infections: critical role of high-molecular weight S-layer proteins in virulence. AB - Wild-type Campylobacter fetus strains possess high-molecular-weight S-layer proteins (S+) and are highly resistant to serum-mediated killing and phagocytosis. Spontaneous mutant strains lacking these proteins (S-) are serum and phagocytosis sensitive and have reduced virulence in a mouse model. Intact S+ cells were treated with pronase, which made them S- although genotypically S+ and had essentially no effect on other cellular proteins or on viability. Treatment with pronase, but not buffer alone, rendered these cells serum and phagocytosis sensitive and reduced mouse virulence to the level observed for the S- mutant cells. In related studies, purified S-layer proteins diminished neutrophil chemoluminescent responses to a heterologous particulate antigen. Finally, passive administration of antiserum to the 97-kDa S-layer protein partially protected mice against lethal challenge with the S+ strain. These studies define the contribution of the S-layer proteins to C. fetus virulence. PMID- 8421172 TI - Therapeutic trial of lipid X in a canine model of septic shock. AB - Three groups of dogs were given lipid X (0, 1, or 10 mg/kg) every 8 h for for seven doses, starting simultaneously with the intraperitoneal placement of Escherichia coli-containing fibrin clots. All animals developed bacteremia, hypotension, and a pattern of decreased left ventricular ejection fraction characteristic of septic shock (P = .01). Survival rates and survival times were not significantly different between treatment groups (P > .2). In a similar experiment, higher doses of lipid X resulted in a significantly decreased survival time compared with concurrent controls (P = .04). Animals receiving lipid X did not differ from controls in serial determinations of temperature, hemodynamic measurements, or laboratory parameters (except serum total protein). Although lipid X has antiendotoxin effects, no benefit could be demonstrated in this antibiotic-treated, gram-negative bacillary-infected model of septic shock. These data do not support a therapeutic role for lipid X in the treatment of gram negative sepsis. PMID- 8421173 TI - Combined multidrug and Mycobacterium w vaccine therapy in patients with multibacillary leprosy. AB - Immunotherapy with Mycobacterium w vaccine was attempted in patients with borderline-borderline, borderline lepromatous (BL), or lepromatous leprosy (LL) to determine whether immunization can hasten recovery and reduce treatment time by invigorating cell-mediated immunity. Mycobacterium w, a nonpathogenic, rapidly growing, atypical mycobacterium, shares a number of common B and T cell determinants with Mycobacterium leprae and Mycobacterium tuberculosis. Patients receiving the vaccine had rapid clinical improvement and accelerated bacteriologic clearance. After treatment with vaccine for 2 years, 13 of 31 BL and LL patients were bacteriologically negative as were 5 of 25 controls. Vaccinated patients had one of two distinct histologic features, either an upgrading in the disease spectrum or complete clearance of granuloma. Some 80% of lepromin conversions were in BL and LL patients who received vaccine versus none and 14.3% of BL and LL controls, respectively. Thirteen of 17 vaccinated LL patients were released from treatment after 2 years in contrast to 2 of 15 controls. PMID- 8421174 TI - Correlates of Leishmania-specific immunity in the clinical spectrum of infection with Leishmania chagasi. AB - Patients from across the spectrum of clinical manifestations of Leishmania chagasi infection were evaluated for in vitro correlates of immunity. Peripheral blood mononuclear cells were assayed for parasite-specific lymphoproliferation, cytokine generation, and the capacity to activate autologous macrophages to kill intracellular amastigotes. Patients with acute kala-azar were generally unreactive in each of these assays. Children with subclinical infection demonstrated relatively low levels of proliferation and interferon-gamma production, but none went on to develop overt kala-azar during the study. Patients evaluated after therapy for kala-azar demonstrated yet higher levels of lymphoproliferation and cytokine generation and produced low but significant levels of cytokines in vitro in response to parasite antigens, but not during the activation of infected macrophages. Finally, peripheral blood mononuclear cells from adults with positive delayed-type hypersensitivity responses and no history of kala-azar showed the broadest reactivity in vitro. These patients' cells generated the largest amounts of activating cytokines in vitro during the activation of autologous macrophages to a leishmanicidal state. PMID- 8421175 TI - Parasitism of epidermal Langerhans cells in experimental cutaneous leishmaniasis with Leishmania major. AB - Murine epidermal Langerhans cells (LC) have been demonstrated to stimulate a vigorous T cell response to Leishmania major, a cause of human cutaneous leishmaniasis. It was therefore of interest to analyze whether LC can take up viable parasites. Epidermal cells were obtained from mouse ear skin for incubation with L. major and subsequent detection of intracellular parasites by cytochemistry. Freshly isolated LC, but not cultured LC, phagocytosed L. major and the uptake was inhibited by antibodies to the complement receptor type 3. Electron microscopic studies revealed the presence of viable amastigotes within LC. Moreover, with double-labeling techniques, L. major-containing LC could also be detected in infected skin. The results demonstrate that LC can internalize L. major. Since the number of organisms per infected LC remained consistently low, the prime task of LC may not be the promotion of parasite spreading but the presentation of L. major antigen to T cells and, thus, the regulation of the cellular immunity during cutaneous leishmaniasis. PMID- 8421176 TI - Characterization of Giardia duodenalis isolates from a waterborne outbreak. AB - Isolates were retrieved from drinking water and from animal and human sources associated with a waterborne outbreak of giardiasis. This is the first report of water-source and epidemic-associated Giardia isolates being adapted to in vitro propagation. Outbreak-associated, non-out-break-associated, and reference isolates were characterized using isoenzyme electrophoresis and pulsed-field gel electrophoresis (PFGE). All outbreak-associated and 2 other isolates were in one of eight zymodemes. The chromosomal complement of the outbreak-associated isolates was relatively homogeneous; this PFGE karyotype was distinguishable from other karyotypes. Overall results of both characterization methods were similar, although PFGE appears to be a more discriminating biotyping technique. Banding patterns of the outbreak-associated Giardia isolates remained the same even though the parasite passed through different hosts during the outbreak. Heterogeneity of isolates was also demonstrated for the first time within a single community not associated with the outbreak. PMID- 8421177 TI - Breast-feeding during primary maternal human immunodeficiency virus infection and risk of transmission from mother to infant. AB - Examination of breast-fed infants of the complete cohort of Australian women whose primary human immunodeficiency virus (HIV) infection occurred postpartum allows for an estimate of risk of transmission of HIV. Ten women with no other risk factors were infected via blood transfusion postpartum. They breast-fed for up to 9 months; 2 of their infants were infected. Another woman, who shared needles for intravenous drug use, seroconverted 6-10 months post-partum. She breast-fed for 14 months. Retrovirus was visualized in the cellular and cell-free fraction of her milk by electron microscopy. Infection in her infant was confirmed at 12 months. Thus, 3 of the 11 babies at risk became infected, providing an estimate of risk of 27% for breast-feeding during primary maternal infection (95% confidence interval, 6-61%). These data establish the association of primary maternal HIV infection and breast-feeding with a high risk of transmission to infants. PMID- 8421178 TI - Effect of sulbactam on infections caused by imipenem-resistant Acinetobacter calcoaceticus biotype anitratus. AB - A recent outbreak of multiresistant strains of Acinetobacter calcoaceticus biotype anitratus was observed mostly, but not exclusively, in the surgical intensive care unit in our hospital. Disk diffusion and microdilution susceptibility studies demonstrated resistance to imipenem, all aminoglycosides, and all individual beta-lactam antibiotics. Only ampicillin plus sulbactam, cefoperazone plus sulbactam, and polymyxin produced zone sizes and MICs in the susceptible ranges. Determination of MICs and MBCs demonstrated that sulbactam was the antimicrobial agent responsible for the killing of these organisms. Nine of 10 patients who were infected with imipenem-resistant Acinetobacter strains and received ampicillin plus sulbactam for > 3 days improved clinically, and in many cases organisms were eradicated from the site of isolation. PMID- 8421179 TI - A massive outbreak of type E botulism associated with traditional salted fish in Cairo. AB - In April 1991, 91 hospitalized patients in Cairo were reported to the Egyptian Ministry of Health with botulism intoxication. To define the spectrum of illness and identify a food vehicle, 45 patients were interviewed and a case-control investigation was conducted among families of 5 hospitalized patients. Clinical specimens and specimens of implicated food were tested for toxin and cultured for Clostridium botulinum. Hospitalized patients had symptoms consistent with botulism; 18 (20%) of 91 reported patients died. Illness was associated with eating faseikh (uneviscerated, salted mullet fish; lower 95% confidence limit of odds ratio = 6.6, P < .001). All 5 case-families purchased faseikh from one shop. Very high levels of type E botulinal toxin were detected in faseikh reported to be purchased from the implicated shop; C. botulinum type E was isolated from cultures of clinical specimens and from the faseikh. This is the first documented outbreak of botulism in Egypt and the largest type E outbreak ever reported. PMID- 8421180 TI - Detection of toxigenic Clostridium difficile in stool specimens by the polymerase chain reaction. AB - Polymerase chain reaction (PCR) amplification of a segment of the toxin A gene was used to detect toxigenic Clostridium difficile directly from stool specimens of patients with antibiotic-associated diarrhea. Although PCR-inhibitory substances were recognized in DNA prepared from stool specimens, the inhibitory substances were eliminated by using an ion-exchange column after phenol chloroform extraction. Eventually, 39 stool specimens were evaluated by PCR. PCR results for detection of toxigenic C. difficile were in complete agreement with cell culture assay results; all 12 PCR-positive stool specimens were positive by cytotoxin assay, and all 27 PCR-negative specimens were negative by cytotoxin assay. Toxigenic C. difficile was cultured from all PCR-positive specimens. These results suggest that PCR amplification may be an effective method for laboratory diagnosis of C. difficile-associated diarrhea and colitis. PMID- 8421181 TI - Detection of Shigellae and enteroinvasive Escherichia coli by amplification of the invasion plasmid antigen H DNA sequence in patients with dysentery. AB - Detection of Shigella organisms and enteroinvasive Escherichia coli (EIEC) by polymerase chain reaction (PCR) was evaluated in 20 patients with dysentery before and in 17 of the 20 after treatment with ciprofloxacin. DNA sequences coding for IpaH antigen, a multiple copy sequence found on the chromosome, and the invasion plasmid locus (ial) was detected after DNA amplification in 13 stools from patients from whom shigellae or EIEC were isolated but not in 21 nondysenteric stools containing other enteric bacteria. Although shigellae or EIEC were not isolated from any patient with dysentery after ciprofloxacin treatment, IpaH and ial sequences were found after PCR amplification in 7 patients after treatment with ciprofloxacin. IpaH sequences alone were detected in 4 patients; DNA augmentation of IpaH in stools in a specific way to identify Shigella or EIEC infection in persons from whom cultures cannot be obtained promptly after the onset of diarrhea or who have received antibiotics. PMID- 8421182 TI - Endotoxin and tumor necrosis factor-alpha-induced interleukin-8 release in humans. AB - Neutrophil recruitment and activation are thought to play an important role in tissue damage observed in septicemia. Interleukin-8 (IL-8) is a small cytokine with important neutrophil-activating and chemoattractant properties. IL-8 release was studied after injection of human volunteers with low doses of either endotoxin (2 ng/kg of body weight) or tumor necrosis factor-alpha (TNF alpha) (50 micrograms/m2). After TNF-alpha injection, IL-8 appeared at 30 min, whereas increased levels were first observed after 90 min in endotoxin-challenged volunteers. Peak levels were measured at 120 min after both endotoxin (192 +/- 193 ng/L) and TNF alpha (500 +/- 236 ng/L) injection. These data indicate that IL 8 is released in humans after injection of endotoxin and TNF alpha and suggest that endotoxin-induced IL-8 release is mediated by TNF alpha. PMID- 8421183 TI - The biologic activities of peptidoglycan in experimental Haemophilus influenzae meningitis. AB - While gram-positive bacterial cell walls are known to incite inflammation, the contribution of gram-negative peptidoglycan to disease has not been characterized. The ability of cell wall, purified peptidoglycan, and soluble peptidoglycan subcomponents from Haemophilus influenzae to provoke inflammation was determined in a rabbit model of meningitis. Haemophilus peptidoglycan, with or without associated proteins, produced brain edema at > or = 0.1 micrograms/mL of cerebrospinal fluid (CSF); leukocytosis and protein accumulation in CSF occurred only at > or = 10.0 micrograms/mL of CSF. Solubilized peptidoglycan was 10-fold more active than intact cell wall. The bioactivity of peptidoglycan from ampicillin-resistant H. influenzae was at least twofold greater than that of ampicillin-sensitive strains. Consistent with these pathologic effects of purified peptidoglycan, ampicillin-induced bacterial lysates in which endotoxin was neutralized induced brain edema and protein influx but little leukocytosis. Thus, peptidoglycan seems to contribute to the pathology of gram-negative meningitis, particularly brain edema. PMID- 8421184 TI - Ceftriaxone no longer predictably cures chancroid in Kenya. AB - Ceftriaxone in a dose of 250 mg given intramuscularly is currently recommended for the treatment of chancroid. Among 133 men in Nairobi, Kenya, with culture proven chancroid, who were treated with ceftriaxone, treatment failed in 35%. Poor outcome was associated with human immunodeficiency virus type 1 seropositivity. Thus, treatment recommendations for chancroid should be reevaluated. PMID- 8421185 TI - Interleukin-8 in serum and cerebrospinal fluid from patients with meningococcal disease. AB - To evaluate the role of interleukin (IL)-8 in meningococcal disease, a solid phase double-ligand ELISA was used to quantitate IL-8 in sera and cerebrospinal fluid (CSF) from patients with meningococcal meningitis, bacteremia, or both with or without septic shock. IL-8 was demonstrated in sera from 28 of 62 patients; levels were significantly higher in patients with septic shock without meningitis (median, 36.1 ng/mL) than in patients with other manifestations (median, < 0.02 ng/mL), and 4 of 5 patients who died had high levels. IL-8 was detected in all 27 CSF samples. Serum IL-8 levels correlated highly significantly with those of IL-6 (r = .83) and tumor necrosis factor (TNF; r = .64), while the correlations between corresponding CSF levels were less pronounced (r = .43 and r = .38, respectively) but still significant. Serum IL-8 levels were highest in patients with a symptom history < 12 h. The elimination rate of IL-8 from serum varied and was similar to that of IL-6 and TNF. IL-8 appears to participate in the complex cytokine network during the initial phase of systemic meningococcal infections. PMID- 8421186 TI - Vibrio infections on the Gulf Coast: results of first year of regional surveillance. Gulf Coast Vibrio Working Group. AB - In 1989, the first year of coordinated Vibrio surveillance in four Gulf Coast states (Alabama, Florida, Louisiana, and Texas), 121 infections were reported. These included 34 V. parahaemolyticus, 30 V. cholerae non-O1, 18 V. vulnificus, 9 V. hollisae, 7 V. alginolyticus, and 7 V. fluvialis. Fourteen patients had primary septicemia, 71 had gastroenteritis, and 29 had wound infections; 7 had other or unknown illnesses. Sixty-six patients were hospitalized, and 9 died. All patients with primary septicemia, but only 17% of those with gastroenteritis, were known to have an underlying illness (P < .001). Among patients for whom data were available, 67% with primary septicemia and 74% with gastroenteritis ate raw oysters in the week before illness began. Of 50 patients with data on where oysters were obtained, 42 (84%) ate them at oyster bars or restaurants. These data provide evidence that in the Gulf Coast region raw oyster consumption is an important cause of Vibrio-associated gastroenteritis among adults without underlying illnesses. PMID- 8421187 TI - Efficacy of acellular pertussis vaccine in young infants. AB - A prospective study of a pertussis outbreak in a residential facility was done. Among 19 residents aged < or = 2 years, 10 children were unimmunized and 9 were immunized with acellular pertussis vaccines. Of the 10 unimmunized children, 7 acquired laboratory-confirmed pertussis (four-fold titer rise, positive culture, or both); of these, 6 developed typical symptoms. Eight of the 9 immunized children acquired laboratory-confirmed infections, and 1 of these developed typical symptoms. No difference in infection was noted between the unimmunized and immunized groups (7/10 vs. 8/9, respectively), but a significant difference was observed in the development of symptoms (6/10 vs. 1/9, P < .05). Also, the isolation rate of Bordetella pertussis seemed to differ between the two groups (6/10 vs. 2/9, P = .12). In this population, acellular pertussis vaccine did not prevent infection with B. pertussis but protected most young infants from the onset of clinical symptoms. PMID- 8421188 TI - A prospective study of the catheter hub as the portal of entry for microorganisms causing catheter-related sepsis in neonates. AB - The hypothesis that catheter-related sepsis (CRS) may be preceded by contamination of the catheter hub was tested in neonates with central venous catheters. Cultures of the catheter hub were obtained three times per week. One hundred thirteen catheters were placed in 88 patients. Of 35 episodes of sepsis, 28 were catheter-related, for a catheter sepsis rate of 1.03/100 catheter-days. CRS occurred in 26 (23%) of 113 catheters. In 10 of 28 episodes, the infecting microorganism was cultured from the hub before its culture from blood obtained at the time of clinical sepsis. In an additional 5 cases, a culture of the catheter hub at the time of clinical sepsis yielded the same isolate as the blood culture. Thus, 54% of episodes of CRS were preceded by or coincided with contamination of the hub. The catheter hub may be a major portal of entry for microorganisms causing sepsis in a neonatal intensive care unit. PMID- 8421189 TI - The activity of atovaquone (566C80) in murine toxoplasmosis is markedly augmented when used in combination with pyrimethamine or sulfadiazine. AB - The activity of atovaquone in the treatment of murine toxoplasmosis was greatly enhanced when administered in combination with pyrimethamine or sulfadiazine. Mice infected with lethal inocula of tachyzoites or cysts of Toxoplasma gondii and treated with doses of atovaquone, pyrimethamine, or sulfadiazine that were ineffective when administered alone had 70% survival when pyrimethamine plus atovaquone and 100% survival when sulfadiazine plus atovaquone was used. Of interest, doses of pyrimethamine and, particularly, sulfadiazine far below the doses that would induce any protection in infected mice were active when combined with atovaquone. These results suggest that clinical trials for treatment of toxoplasmosis in AIDS patients using the combination of atovaquone with sulfadiazine or pyrimethamine are justified. PMID- 8421190 TI - Cell-mediated immune response to human immunodeficiency virus in homosexual men. PMID- 8421191 TI - Intravenous immune globulin for passive tetanus prophylaxis. PMID- 8421192 TI - Possible association of HLA-DR2 phenotype and detectable human immunodeficiency virus (HIV) p24 antigen in HIV-positive patients. PMID- 8421193 TI - Ground beef consumption in noncommercial settings is a risk factor for sporadic Escherichia coli O157:H7 infection in Canada. PMID- 8421194 TI - Persistence of Chlamydia complement-fixation antibody after an outbreak of psittacosis. PMID- 8421195 TI - Increased interferon-gamma and neopterin concentrations in patients with acute brucellosis. PMID- 8421196 TI - Levels of interleukin-8 in patients with adult respiratory distress syndrome. PMID- 8421197 TI - Occlusive hyperemia: a new way to think about an old problem. PMID- 8421198 TI - Occlusive hyperemia: a theory for the hemodynamic complications following resection of intracerebral arteriovenous malformations. AB - An alternative theory is proposed to explain the brain edema and hemorrhage that may occur after resection of high-flow intracerebral arteriovenous malformations (AVM's). This theory, termed "occlusive hyperemia," is based on a retrospective analysis of operative dictations along with postoperative imaging studies (191 angiograms and 273 computerized tomography scans) in 295 cases of intracerebral AVM's operated on at the Mayo Clinic between 1970 and 1990. In this series, 34 cases (12%) of postoperative deterioration were documented, of which 15 were due to incomplete resection of the AVM. Of the remaining 19 cases, six had brain edema alone and 13 had hemorrhage with edema, despite complete excision of the AVM. In these 19 cases, the AVM's were greater than 6 cm in diameter in 10 patients, between 3 and 6 cm in six, and less than 3 cm in three. Obstruction of the venous drainage system was observed in 14 (74%) of the 19 cases. Ten of these 14 were due to obstruction of the primary venous drainage of the brain parenchyma immediately surrounding the lesions, while four were due to obstruction of other venous structures. In no case was a rapid circulation identified on postoperative angiograms. The flow pattern was slow or stagnant in former AVM feeders and their parenchymal branches. It is proposed that postoperative intracranial hemorrhage and/or brain edema in AVM patients may be due to: 1) obstruction of the venous outflow system of brain adjacent to the AVM, with subsequent passive hyperemia and engorgement; and 2) stagnant arterial flow in former AVM feeders and their parenchymal branches, with subsequent worsening of the existing hypoperfusion, ischemia, and hemorrhage or edema into these areas. Supportive hemodynamic evidence for this theory was derived from the literature. PMID- 8421199 TI - Complications of surgery for arteriovenous malformations of the brain. AB - A series of 112 patients undergoing complete surgical resection of arteriovenous malformations (AVM's) of the brain between 1974 and 1990 were analyzed for complications and 12-month outcomes. The cohort consisted of 44 patients with small AVM's (< 2 cm in diameter), 43 patients with medium-sized AVM's (2 to 4 cm in diameter), and 25 patients with large AVM's (> 4 cm in diameter). There was a 3.6% series mortality rate and an 18% morbidity rate. One of the four deaths was caused by normal perfusion pressure breakthrough. Analysis of logistic regression found that the most important factor influencing the occurrence of complications in this series was AVM size (p = 0.005) and that the occurrence of complications (p < 0.001) and the neurological grade at the time of surgery (p < 0.004) both significantly contributed to the outcome at 12 months. This study stresses the importance of defining complications in terms of rigid criteria when analyzing AVM series in order to allow for a correct evaluation of the risk:benefit ratio of surgery. Furthermore, it emphasizes the need for a separate analysis of the importance of complications upon outcome. PMID- 8421200 TI - Use of transcranial Doppler sonography to predict development of a delayed ischemic deficit after subarachnoid hemorrhage. AB - Blood flow velocity was recorded from the middle or anterior cerebral and extracranial internal carotid arteries using transcranial Doppler sonography (TCD) in 121 unselected consecutive patients with acute aneurysmal subarachnoid hemorrhage (SAH). Recordings were made daily or every 2nd day after SAH for a 14 day period. The highest recorded velocity was greater in the 47 patients who developed a delayed ischemic neurological deficit (186 +/- 6 cm sec-1; mean +/- standard error of the mean) than in the 74 patients who did not develop a neurological deficit (149 +/- 5 cm sec-1) (p < 0.001, Mann-Whitney test). Peak velocity recordings can thus assist in the diagnosis of delayed ischemic neurological deficit; however, peak velocity was often recorded only after the onset of neurological deficit. When only those readings made before the onset of neurological deficit were considered, there was no significant difference in peak velocity between the groups (157 +/- 8 cm sec-1 vs. 149 +/- 5 cm sec-1, respectively). Alternative TCD parameters for predicting delayed neurological deficit were therefore sought. The rate of increase in TCD velocity, recorded during the first few days after SAH, was significantly higher in the patients who later developed a neurological deficit. A maximum velocity increase of 65 +/- 5 cm sec-1 per 24-hour period was recorded in patients who later developed a neurological deficit, compared to 47 +/- 3 cm sec-1 24 hrs-1 in patients who did not develop a delayed neurological deficit (p = 0.003). A rise of more than 50 cm sec-1 24 hrs-1 identifies those patients who are most likely to develop a delayed ischemic neurological deficit after SAH. This can be applied prospectively to individual cases. Serial TCD studies in the early period after SAH are thus of value to identify patients who can be selected for prophylactic therapy, which may prevent or ameliorate development of delayed ischemic neurological deficits. PMID- 8421201 TI - Intracerebral hemorrhage more than twice as common as subarachnoid hemorrhage. AB - The authors report a study of all instances of spontaneous intracerebral hemorrhage (ICH) (188 cases) and subarachnoid hemorrhage (SAH) (80 cases) that occurred in the Greater Cincinnati area during 1988. Adjusted for age, sex, and race, the annual incidence of ICH was 15 per 100,000 population (95% confidence interval 13 to 17) versus six per 100,000 for SAH (95% confidence interval 5 to 8). The incidence of ICH was at least double that of SAH for women, men, and whites and approximately 1 1/2 times that for blacks. The 30-day mortality rate of 44% for ICH was not significantly different from the 46% mortality rate for SAH. Despite the evidence that ICH is more than twice as common and the disorder just as deadly as SAH, clinical and laboratory research continues to focus primarily on SAH. PMID- 8421202 TI - Transluminal angioplasty for atherosclerotic disease of the vertebral and basilar arteries. AB - Transluminal angioplasty for hemodynamically significant stenosis (> 70%) involving the posterior cerebral circulation is now being performed by the authors in selected cases. A total of 42 lesions affecting the vertebral or basilar artery have been successfully treated by percutaneous transluminal angioplasty techniques in 41 patients. The lesions involved the proximal vertebral artery in 34 cases, the distal vertebral artery in five, and the basilar artery in three. Patients were examined clinically at 1 to 3 and 6 to 12 months after angioplasty. Three (7.1%) permanent complications occurred, consisting of stroke in two cases and vessel rupture in one. There were four (9.5%) transient complications (< 30 minutes): two cases of vessel spasm and two of cerebral ischemia. Clinical follow-up examination demonstrated improvement of symptoms in 39 cases (92.9%). Radiographic follow-up studies demonstrated three cases (7.1%) of restenosis involving the proximal vertebral artery; two were treated by repeat angioplasty without complication, and the third is being followed clinically while the patient remains asymptomatic. In patients with significant atherosclerotic stenosis involving the vertebral or basilar artery territories, transluminal angioplasty may be of significant benefit in alleviating symptoms and improving blood flow to the posterior cerebral circulation. PMID- 8421203 TI - Total calvarial reconstruction for sagittal synostosis in older infants and children. AB - Premature closure of the sagittal suture is the most common form of craniosynostosis, but this condition occasionally goes unrecognized until the child is too old to undergo procedures that depend upon continued calvarial growth for success. As the entire calvaria is affected and thus misshapen by sagittal synostosis, late correction involves total calvarial reconstruction. The extensive nature of this undertaking has precluded its utilization despite the presence of significant deformities. Adapting the techniques and experience gained from craniofacial surgery, the authors performed total calvarial reconstruction on nine children with sagittal synostosis and subsequent scaphocephaly diagnosed after the age of 1 year. In each case the goals of shortening the anteroposterior length, widening the biparietal diameter, and reducing frontal and occipital deformities were met. Morbidity consisted of acute blood loss, postoperative hyponatremia, and in one case a residual skull defect. The rationale for this procedure and the techniques utilized are discussed. PMID- 8421204 TI - Transsphenoidal adenomectomy for growth hormone-secreting pituitary adenomas in acromegaly: outcome analysis and determinants of failure. AB - The results of transsphenoidal adenomectomy for growth hormone (GH)-secreting pituitary adenomas in acromegaly performed over a 17-year period were analyzed retrospectively to determine which preoperative factors significantly influenced the long-term surgical outcome. These variables were then used to develop a logistic regression model to determine the probability of surgical failure. The series consisted of 103 patients. Long-term follow-up study (mean duration 102 +/ 64 months) was performed to derive outcome analysis and determinants of failure. Surgical control was defined as a long-term postoperative serum basal GH level of less than 5 micrograms/liter, a long-term postoperative serum somatomedin C (SM C) level of less than 2.2 U/ml, and a favorable clinical response. Eighteen (17.5%) patients did not meet these criteria. The overall control rate by the GH criteria was 81.3% and by the SM-C criteria 76.2%. By multivariate logistic regression analysis, tumor stage was the strongest predictor of outcome (p < 0.05). The preoperative GH level, tumor grade, and preoperative SM-C level were significant univariate predictors (p < 0.05). There were statistically significant differences in mean preoperative GH and SM-C levels (p < 0.05, t test) and tumor stage (p < 0.05, chi-squared test) between patients whose acromegaly was controlled by surgery and those whose acromegaly was not. Furthermore, estimates were derived of the probability of surgical failure based on preoperative GH level, preoperative SM-C level, and tumor grade and stage. The authors believe these findings will enhance clinical decision-making for neurosurgeons considering transsphenoidal microsurgery in patients with acromegaly. PMID- 8421205 TI - Intrathecal baclofen for intractable spasticity of spinal origin: results of a long-term multicenter study. AB - A total of 93 patients with intractable spasticity due to either spinal cord injury (59 cases), multiple sclerosis (31 cases), or other spinal pathology (three cases) were entered into a randomized double-blind placebo-controlled screening protocol of intrathecal baclofen test injections. Of the 88 patients who responded to an intrathecal bolus of 50, 75, or 100 micrograms of baclofen, 75 underwent implantation of a programmable pump system for chronic therapy. Patients were followed for 5 to 41 months after surgery (mean 19 months). No deaths or new permanent neurological deficits occurred as a result of surgery or chronic intrathecal baclofen administration. Rigidity was reduced from a mean preoperative Ashworth scale score of 3.9 to a mean postoperative score of 1.7. Muscle spasms were reduced from a mean preoperative score of 3.1 (on a four-point scale) to a mean postoperative score of 1.0. Although the dose of intrathecal baclofen required to control spasticity increased with time, drug tolerance was not a limiting factor in this study. Only one patient withdrew from the study because of a late surgical complication (pump pocket infection). Another patient received an intrathecal baclofen overdose because of a human error in programming the pump. The results of this study indicate that intrathecal baclofen infusion can be safe and effective for the long-term treatment of intractable spasticity in patients with spinal cord injury or multiple sclerosis. PMID- 8421206 TI - Mapping of sensory responses to epidural stimulation of the intraspinal neural structures in man. AB - A database is presented of sensory responses to electrical stimulation of the dorsal neural structures at various spine levels in 106 subjects subjected to epidural spinal cord stimulation. All patients were implanted for chronic pain management and were able to perceive stimulation in the area of pain. All patients entered in this study were able to reliably report their stimulation pattern. Several patients were implanted with more than one electrode array. The electrode arrays were placed in the dorsal epidural space at levels between C-1 and L-1. The structures that were likely involved include the dorsal roots, dorsal root entry zone, dorsal horn, and dorsal columns. At the present time, exact characterization of the structure being stimulated is possible only in limited instances. Various body areas are presented with the correspondent spine levels where implanted electrodes generate paresthesias. Areas that are relatively easy targets for stimulation are the median aspect of the hand, the abdominal wall, the anterior aspect of the thigh, and the foot. Some areas are particularly difficult to cover with stimulation-induced paresthesias; these include the C-2 distribution, the neck, the low back, and the perineum. PMID- 8421207 TI - Reinforced cytotoxicity of lymphokine-activated killer cells toward glioma cells by transfection with the tumor necrosis factor-alpha gene. AB - Lymphokine-activated killer (LAK) cells generated from peripheral blood lymphocytes incubated with recombinant interleukin-2 were transfected with the human tumor necrosis factor (TNF)-alpha gene by means of novel liposomes with a positive change on their surface. The cells secreted significant amounts of TNF alpha into the culture medium and exhibited reinforcement of cytotoxicity toward a human glioma cell line (U251-SP), being three times more cytotoxic than nontransfected LAK cells. The mechanism for the reinforcement of cytotoxicity is considered to involve not only an increase in TNF-alpha secretion from LAK cells but also its expression on their surface. Intratumoral or intrathecal injection of LAK cells transfected with the TNF-alpha gene may be useful for the treatment of patients with malignant gliomas. PMID- 8421208 TI - Contributions of ions and albumin to the formation and resolution of ischemic brain edema. AB - Changes in brain water, sodium, potassium, and albumin contents and blood-brain barrier (BBB) permeability were determined at various times between 1 hour and 6 weeks following occlusion of the middle cerebral artery (MCA) in rats. In the center of the infarct, brain edema increased to a maximum level by 12 hours, remained elevated for 7 days, and then returned to normal. The change in water content was accompanied by a parallel increase in sodium and decrease in potassium contents; however, the increase in sodium always exceeded the decrease in potassium, resulting in a net gain in brain cations during edema formation which returned to normal with edema resolution. The BBB permeability to 3H-alpha aminoisobutyric acid was increased by 24 hours after MCA occlusion and returned to normal by 1 week after the edema had resolved. The time course for changes in brain albumin content was different than that for brain edema formation. Large increases in brain albumin content were not apparent until 6 hours after the onset of ischemia, rose to a peak at 3 days after occlusion of the MCA, and returned to normal several weeks after the edema had resolved. Albumin appeared to spread from the central infarct zone to surrounding, less ischemic areas. The relative contributions of the osmotic force produced by the increase in brain cations and the oncotic force produced by the increase in brain albumin to the observed change in water content were calculated. At all time points, the increase in brain cations accounted for nearly all of the observed brain edema, while the increase in albumin played essentially no role in edema development. PMID- 8421209 TI - Local protective effects of nerve growth factor-secreting fibroblasts against excitotoxic lesions in the rat striatum. AB - Neurotrophic factors, such as nerve growth factor (NGF), in addition to their role in neuronal development, have protective effects on neuronal survival. Intracerebral implantation of cells genetically altered to secrete high levels of NGF is also found to promote neuronal survival in experimental lesioning models of the brain. The range of activity for such biological delivery systems has not yet been well described either spatially or temporally. Therefore, the authors chose to study the local and distant protective effects of an NGF-secreting rat fibroblast cell line implanted in an excitotoxic lesion model of Huntington's disease. They found that preimplantation of NGF-secreting fibroblasts placed within the corpus callosum reduced the maximum cross-sectional area of a subsequent excitotoxic lesion in the ipsilateral striatum by 80% when compared to the effects of a non-NGF-secreting fibroblast graft, and by 83% when compared to excitotoxic lesions in ungrafted animals (p < 0.003). However, NGF-secreting cells placed in the contralateral corpus callosum failed to affect striatal lesion size significantly when compared to contralateral or ipsilateral non-NGF secreting cell implants. Of note, fibroblasts were clearly visible within the graft site at 7 and 18 days after implantation; however, few cells within the grafts stained positively for NGF peptide or for the messenger ribonucleic acid (mRNA) encoding the transfected NGF gene-construct at either time point. These results show that biological delivery systems for NGF appear to have a profound but local effect on neuronal excitotoxicity, which will necessitate careful neurosurgical placement for maximum effect. Furthermore, the ability of this genetically altered cell line to synthesize NGF mRNA and peptide appears to decrease spontaneously in vivo, a characteristic that will need to be addressed before this method of biological delivery can be utilized as a treatment for chronic degenerative diseases. PMID- 8421210 TI - Noncommunicating syringomyelia following occlusion of central canal in rats. Experimental model and histological findings. AB - This report describes a new and reliable technique for producing experimental noncommunicating syringomyelia. In 30 rats, 1.2 to 1.6 microliters of kaolin was microinjected into the dorsal columns and central gray matter of the spinal cord at C-6. The inoculations caused transient neurological deficits in four animals and no deficits in 26 animals. Within 24 hours, kaolin and polymorphonuclear leukocytes entered the central canal and drained rostrally. The clearance of inflammatory products induced a proliferation of ependymal cells and periependymal fibrous astrocytes, which formed synechiae and obstructed the canal at the level of injection and at one or more levels up to C-1. In 22 animals followed for 48 hours or longer, the upper end of the central canal became acutely dilated and formed an ependyma-lined syrinx that enlarged to massive dimensions within 6 weeks. The rostral syrinxes did not communicate with the fourth ventricle and were not associated with hydrocephalus. The histological findings in acute noncommunicating syringomyelia were characterized by progressive stretching and thinning of the ependyma, elongation of intracanalicular septae, and the formation of periependymal edema. After 3 weeks, there was progressive compression of the periependymal tissues associated with stretching of axons, fragmentation of myelin sheaths, and the formation of myelin droplets. These findings and the sequence in which they evolved were identical in most respects to those occurring in acute and subacute noncommunicating hydrocephalus. PMID- 8421211 TI - Spinal angiolipomas. Report of three cases. AB - Spinal angiolipomas are distinct, benign lesions composed of mature lipocytes admixed with abnormal blood vessels. Three new cases of spinal angiolipoma are presented and 34 previously reported cases are analyzed. The 37 total cases (23 females and 14 males) ranged in age from 17 to 73 years (mean 43 years; median 45 years). The mean age of the female patients was older than that for the males (45.0 vs. 41.6 years; p < 0.001, Student's t-test) and most were peri- or postmenopausal. Prior to diagnosis, 97% of the patients had weakness of the lower extremities, 94% had sensory dysfunction, 84% had hyperreflexia and spasticity, 51% had sphincter dysfunction, and 41% had back pain lasting from 1 to 180 months (mean 28 months). Five (22%) of the 23 female patients were pregnant and two had exhibited significant weight gain coincident with the onset of symptoms. The angiolipomas were extradural in 35 patients and intramedullary in two; seven of the extradural lesions infiltrated the surrounding bone. The tumors extended from C-6 to L-4 and had a predilection for the midthoracic region (53% of cases). Plain radiographs were abnormal in 11 (39%) of 28 patients and in all patients with bone infiltration. Myelograms were abnormal in 97% of 32 patients and showed a complete block in 63% of patients. Computerized tomography (CT) and magnetic resonance (MR) imaging revealed the fat-density lesions in all cases studied. There was vascular enhancement in three of five cases with contrast-infused CT and in the one case with gadolinium-infused MR imaging. All patients improved following resection of the epidural lesions and internal decompression of the intramedullary lesions. It is concluded that spinal angiolipomas predominantly affect women. They involve the thoracic (especially the midthoracic) region, and produce symptoms and signs of spinal compression and, in some cases, bone erosion and pathological fractures. Their symptomatology can be exacerbated by pregnancy and weight gain, suggesting that vascular engorgement and the presence of obesity influence their evolution. Their preponderance in older, peri-, or postmenopausal women, and their clinical exacerbation in pregnant women support a role for hormonal influence. Magnetic resonance imaging is the investigation of choice for the diagnosis of these lesions. Surgery is universally successful in relieving symptoms. PMID- 8421212 TI - Childhood meningioma associated with meningio-angiomatosis. Case report. AB - This paper reports childhood meningioma in association with meningioangiomatosis. The patient was an 11-month-old baby boy who presented with a left focal seizure. He had no stigmata of neurofibromatosis. Computerized tomography and magnetic resonance imaging revealed an extra-axial, contrast-enhancing mass in the interhemispheric fissure which indented the right frontal lobe. The tumor was totally removed. Microscopically, the lesion was a fibrous and transitional meningioma with foci of necrosis and scattered mitotic figures. The adherent neural parenchyma showed the histological features of meningioangiomatosis. It is concluded that meningioangiomatosis may accompany childhood meningiomas more often than is generally appreciated. PMID- 8421213 TI - Contralateral external carotid-to-middle cerebral artery graft using the saphenous vein. Case report. AB - A variation of the extracranial-intracranial arterial bypass, using a long saphenous vein graft, is presented. The saphenous vein graft was inserted from the contralateral external carotid artery to the distal middle cerebral artery to replace the common and internal carotid arteries in a patient with a large neck tumor that invaded the common and internal carotid arteries, the esophagus, and the trachea. The patient had a positive balloon Matas' test. The saphenous vein was covered with an artificial vascular graft so that turning of the head or movement of the mandible did not displace or compress the graft. A large volume of flow began immediately after anastomosis. A description of the case and the operative technique is presented herein. PMID- 8421214 TI - Intracerebral polyposis. Case report. AB - A 25-year-old man presented with nontraumatic cerebrospinal fluid rhinorrhea and meningitis. On investigation, he was found to have a multiloculated intracerebral cystic lesion of the right frontal lobe with a bony lesion inside the cyst, just above the right cribriform plate. Surgery revealed multiple grape-like cystic pedunculated lesions with narrow stalks attached to a bony outgrowth which was adherent to the right cribriform plate. Macroscopically and microscopically, the excised lesions were similar to nasal polyps. PMID- 8421215 TI - Intracranial xanthogranuloma of the dura in Hand-Schuller-Christian disease. Case report. AB - The authors describe a 37-year-old man with the classic clinical features of Hand Schuller-Christian disease. He presented with symptoms of increased intracranial pressure due to obstructive hydrocephalus secondary to a huge xanthogranuloma involving falx cerebri and tentorium cerebelli. Immunohistochemical and ultrastructural studies failed to demonstrate Langerhans histiocytes, however. The implication of this finding is discussed in light of the recent relevant literature. PMID- 8421216 TI - The Duke University Medical Center. AB - The events leading up to the creation of Duke University, the Duke University School of Medicine, and Duke Hospital are reviewed. The efforts of many individuals during more than 80 years were rewarded by an endowment and then a bequest by James B. Duke that converted Trinity College into Duke University and made possible the origination of its Medical Center. The first neurosurgical operation at the new hospital was performed on July 24, 1930, the fourth day it was open. PMID- 8421217 TI - Neurogenic hypertension. PMID- 8421218 TI - Thalamic target for tremor. PMID- 8421219 TI - Spinal accessory-facial nerve anastomosis. PMID- 8421220 TI - Anomalies of the A1 segment. PMID- 8421221 TI - Long-chain dietary fatty acids affect the capacity of Drosophila melanogaster to tolerate ethanol. AB - Four-day post-hatch larvae (mid-third instar) of Drosophila melanogaster were fed an intermediate diet with or without supplement of an individual fatty acid for 2 d and then transferred to a diet with a growth-limiting level of 0.94 mol/L ethanol (5.5%, v/v) or an ethanol-free diet. The ethanol stress decreased survival and larval development rate but increased the weight of surviving adult males. Dietary long-chain fatty acids altered the fatty acid composition of tissue lipids of larvae. When an unsaturated fatty acid was fed, except for 18:2(n-6), the tissue level of total unsaturated fatty acids was markedly increased. Both saturated and unsaturated 18-carbon fatty acids shortened larval development time. Linoleic acid [18:2(n-6)] and linolenic acid [18:3(n-3)] enhanced survival overall and, together with stearic acid (18:0), gave marked protection from ethanol stress in terms of survival. Correlation analysis across the different fatty acid diets indicated a strong positive association between tissue 18-carbon fatty acid levels and ethanol tolerance and between 18-carbon fatty acid levels and development rate. No major differences were observed in the effects of the fatty acids on the Canton-S and OD4 (Tahbilk) wild-type strains. Thus, the fatty acid content of D. melanogaster larvae is important for growth and survival in ethanol-rich habitats. PMID- 8421222 TI - Intestinal solubility and absorption of ferrous iron in growing rats are affected by different dietary pectins. AB - The effects of pectin structure on iron solubility were examined in vivo and in vitro using pectins differing in degree of esterification (DE) and molecular weight (MW). The pectins prepared differed (in DE and MW, respectively) as follows: P-A (73%, 860,000), P-B (75%, 89,000), P-C (22%, 1,260,000) and P-D (24%, 114,000). Weanling rats were trained to consume a basal diet containing ferrous sulfate as the iron source within 2 h. Food-deprived rats were presented the basal diet or the same diet containing one of the pectins at 80 g/kg diet. One hour after the meal, soluble, insoluble and tissue iron in intestinal segments and serum iron concentration in portal blood were determined and amounts of absorbed iron were calculated. Portal blood iron and calculated absorbed iron were highly correlated (r = 0.97). None of the pectins reduced iron solubility or absorption below that in rats fed the basal diet. Addition of P-B increased the amount of iron absorbed and postprandial serum iron concentration in portal blood. Solubility of ferrous sulfate in vitro was higher in solutions containing pectins than in a cornstarch solution (P < 0.05) and was also higher in diets containing pectin than in the basal diet (P < 0.05). The pectin with the high DE and low MW (P-B) improved iron solubility and absorption of solubilized iron. PMID- 8421223 TI - Requirements of juvenile milkfish (Chanos chanos Forsskal) for essential amino acids. AB - The dietary requirements of juvenile milkfish (Chanos chanos Forsskal) for essential amino acids were determined in a series of experiments. The fish (< or = 8.0 g) were reared in fiber glass tanks provided with flow-through seawater at 28 degrees C and salinity of 32 g/L for 12 wk. In each experiment, a series of amino acid test diets was formulated containing a combination of intact protein sources (casein-gelatin, fish meal-gelatin, fish meal-soybean meal or fish meal zein) and crystalline amino acids to simulate the levels found in milkfish tissue proteins except for the test amino acid. Each set of isonitrogenous diets contained 40-45% protein and graded levels of the amino acid to be tested. At the end of the feeding experiment, growth, survival and feed efficiency were determined. The requirement level for each essential amino acid was estimated from breakpoint analysis of the growth curve. The dietary essential amino acid requirements (as the percentage of dietary protein) of milkfish juveniles were as follows: arginine, 5.25; histidine, 2.00; isoleucine, 4.00; leucine, 5.11; lysine, 4.00; methionine, 2.50 (cystine, 0.75); phenylalanine, 4.22 (tyrosine, 1.00) or 2.80 (tyrosine, 2.67); threonine, 4.50; tryptophan, 0.60; valine, 3.55. This information is valuable in developing cost-effective practical or commercial feeds and research diets for milkfish juveniles. PMID- 8421224 TI - Dietary zinc affects serum concentrations of insulin and insulin-like growth factor I in growing lambs. AB - Glucose tolerance and concentrations of insulin, somatotropin, insulin-like growth factor-I and cortisol were evaluated in lambs deficient, marginal or adequate in zinc. There were three treatments: 1) deficient (basal diet deficient in zinc; 3.7 mg zinc/kg diet); 2) marginal (basal diet + 5 mg zinc/kg diet); and 3) adequate (basal diet + 40 mg zinc/kg diet). Lambs fed the zinc-deficient diet had lower (P < 0.05) serum insulin concentrations 1 h after feeding compared with those fed the marginal diet, whereas the concentrations in lambs fed the adequate diet were intermediate. Dietary zinc did not affect plasma glucose or serum somatotropin before or after feeding or intravenous glucose administration. A growth hormone-releasing factor analog was given to evaluate concentrations of somatotropin. Serum somatotropin in response to growth hormone-releasing factor analog tended to be higher (P = 0.20) in deficient and marginal lambs when compared with adequate lambs. Serum insulin-like growth factor-I was lower (P < 0.05) in deficient lambs than in marginal or adequate lambs. Cortisol concentrations were not affected (P > 0.05) by zinc status. Severe zinc deficiency altered circulating concentrations of insulin, insulin-like growth factor-I and somatotropin, whereas a marginal deficiency had no effect in growing lambs. PMID- 8421225 TI - Influence of dietary calcium and vitamin D on colon epithelial cell proliferation and 1,2-dimethylhydrazine-induced colon carcinogenesis in rats fed high fat diets. AB - We investigated whether increased levels of dietary calcium and vitamin D could inhibit colon carcinogenesis in rats injected with a single dose of 1,2 dimethylhydrazine. Rats were given a single subcutaneous injection (200 mg/kg body wt) 2 wk before they were fed purified diets containing 20% fat for 32 wk. Diets contained one of three levels of calcium (5, 10 or 15 g/kg diet) as calcium gluconate and one of three levels of vitamin D (0.025, 0.05 or 0.1 mg/kg diet) as cholecalciferol in a 3 x 3 factorial design. Rats receiving the highest level of vitamin D had greater plasma concentrations of 25-hydroxy-vitamin D. Autoradiographic examination of [3H]thymidine-treated rats demonstrated that a higher dietary level of calcium as well as higher levels of vitamin D significantly affected cellular kinetic indices. The total tumor incidence and tumor incidence in the distal colon was 45% lower in rats fed the highest level of both calcium and vitamin D compared with the other eight groups, although this decrease was not statistically significant (P = 0.12). The possible importance of these observations is discussed. PMID- 8421226 TI - Brainstem auditory evoked potential interwave intervals are prolonged in vitamin B-6-deficient cats. AB - Vitamin B-6 deficiency has been reported to produce behavioral, neurophysiological and neuropathological abnormalities in a variety of species. In this investigation we used brainstem auditory evoked potentials (BAEP) to determine if vitamin B-6 deficiency in cats affected peripheral and brainstem auditory pathways. Brainstem auditory evoked potentials were recorded from growing cats as they developed vitamin B-6 deficiency, which was confirmed using clinical, hematological and urinary criteria. The BAEP interwave intervals measured from early (wave 1 or 1N) to late waves (5N) or from middle (wave 3) to late waves increased significantly, whereas interwave intervals from early to middle waves did not differ significantly. These results indicate that vitamin B 6 deficiency affects one or more structures of the brainstem that generate the later parts of the BAEP. The finding of prolonged interwave intervals in vitamin B-6-deficient animals is consistent with slowed axonal conduction velocity secondary to defective myelination. Recording BAEP provided a noninvasive means of detecting effects of vitamin B-6 deficiency on specific parts of the central nervous system. PMID- 8421227 TI - Methionine but not folinic acid or vitamin B-12 alters the frequency of neural tube defects in Axd mutant mice. AB - The Axd (axial defects) mutation, which causes open neural tube defects (NTD) in 25-30% of d 14 mouse embryos of heterozygous (Axd/+) matings and curly tails (CT) in 31%, can be useful in studying neurovertebral morphogenesis. Current interest focuses on the role of methionine in neurulation, because supplementation of dams with this essential amino acid (70 mg/kg body wt) has been shown to reduce by 41% the incidence of NTD among embryos of Axd/+ x Axd/+ crosses when administered on d 8 and 9 of pregnancy. The present experiments were undertaken to determine whether supplementation of dams with a higher dose of methionine could effect a greater level of remediation. At a dose of 180 mg/kg body wt, the amino acid produced a reduction in NTD of 47%, similar to that produced at 70 mg/kg body wt. Supplementation of dams of reciprocal matings of Axd/+ x BALB/cByJ or CF-1 mice allowed assessment of the effects of methionine in heterozygous embryos, which exhibit CT. The amino acid had no effect on Axd/+ embryos from the BALB crosses, but the frequency of CT declined significantly among embryos of the CF-1 outcrosses. Maternal supplements of folinic acid (33 mg/kg) or vitamin B-12 (330 mg/kg) did not alter the incidence of NTD or CT among Axd embryos. No difference in methionine concentration was detected in the serum of Axd/+ and +/+ dams. PMID- 8421228 TI - Cloning of rat intestinal mRNAs affected by zinc deficiency. AB - Male rats were fed a purified diet containing 1 mg Zn/kg to induce zinc deficiency (-Zn). A second set of rats were fed a 30 mg Zn/kg diet, which supplied adequate levels of zinc (+Zn). The zinc-adequate rats were pair-fed to the rats fed the 1 mg Zn/kg diet to eliminate differences in diet intake. After 16 d, the two conditions of dietary zinc status were confirmed by assaying serum zinc concentrations and by Northern analysis for metallothionein mRNA abundance in the kidney. Messenger RNA was purified from small intestine by oligo(dT) chromatography and pooled within conditions. A cDNA plasmid library was constructed from the mRNA derived from the intestines of zinc-deficient rats. The library was then screened by the method of differential hybridization using 32P labeled first strand cDNA probes derived from the +Zn and -Zn mRNA. After screening 10,000 independent cDNA clones, nine cDNAs were isolated and studied further, corresponding to mRNAs that are down-regulated by zinc deficiency. The ratio of the abundance between the two conditions for the nine mRNAs ranged from 1.5- to sevenfold as determined by Northern analysis of the +Zn and -Zn intestinal mRNAs. Two of these clones seemed to be specific to the intestine, and four others were abundant in the intestine and one or two other tissues. We are using these cDNAs as models to study gene regulation under various conditions of dietary zinc intake and to explore the genes most sensitive to zinc deficiency. PMID- 8421229 TI - Restriction of maternal dietary carbohydrate decreases fetal brain indoles and glycogen in rats. AB - In spite of evidence that dietary carbohydrate can increase brain tryptophan and 5-hydroxytryptamine in adult rats, the possible influence of maternal dietary carbohydrate on fetal brain indoles has received little attention. We studied the effect of graded levels (0, 4, 12 and 60%) of maternal dietary fructose or glucose fed throughout pregnancy on fetal brain glycogen and indoles. The diets were isoenergetic and met the NRC energy requirements for pregnant rats. The results demonstrated that low maternal dietary carbohydrate, with adequate energy intake, reduced fetal brain weight and concentrations of glycogen, tryptophan, 5 hydroxytryptamine and 5-hydroxyindoleacetic acid. There were no significant differences between glucose and fructose feeding at any dietary carbohydrate level for any fetal brain measurements, showing that it was the level and not the type of dietary carbohydrate that was important. Significant correlations between fetal brain 5-hydroxytryptamine and brain glycogen, and between fetal brain 5 hydroxytryptamine and brain weight, suggested that lowered brain 5 hydroxytryptamine was only one symptom of disrupted brain development in fetuses of dams fed low levels of carbohydrate. The results show that dietary carbohydrate restriction during pregnancy can have adverse effects on fetal brain development, glycogen levels, and neurotransmitter synthesis even when maternal dietary protein and energy intake are adequate. PMID- 8421230 TI - Age-related changes in rat adipose tissue cellularity are altered by dietary restriction and exercise. AB - This study examined the effects of diet restriction and exercise training during aging on adipocyte size and number in male Wistar rats. An average of 30 weanling rats each were assigned to one of four experimental groups: ad libitum access to food, nonexercised; diet restricted, nonexercised; ad libitum access to food, exercised; diet restricted, exercised. Diet-restricted groups were allowed free access to food every other day, and, over the course of the study, they consumed 18-25% less than the groups allowed ad libitum access. Exercise training consisted of swimming on alternate days for up to 3 h per exercise period. Fat cell size, number and fat depot mass were determined in the inguinal, epididymal and perirenal fat depots at 12 mo of age and every 4 mo thereafter until 28 mo of age. Fat cell number was not influenced by age, exercise or food restriction in the epididymal or perirenal fat depots. Inguinal cell number was lower (P < 0.05) in the diet-restricted, exercised group at all ages compared with all other treatments. Dietary restriction and exercise training significantly decreased fat depot mass and fat cell size, with dietary restriction having the greatest effect. A bimodal distribution of adipocytes was observed in the epididymal, perirenal and inguinal depots of animals allowed ad libitum access to food. The presence of a large population of small fat cells (< 35 microns) indicated that even in the very aged animal there was the propensity for large variations in fat pad mass. The influence of diet restriction, exercise and aging on adipocyte cell size was highly variable and these effects were dependent on adipose tissue depot location. PMID- 8421231 TI - Dietary sorbose prevents and improves hyperglycemia in genetically diabetic mice. AB - The effect of dietary sorbose on the prevention (Experiment 1) and amelioration (Experiment 2) of diabetes was investigated in the genetically diabetic mouse [C57BL/KsJ (db/db)] for 6 wk. When sucrose (200 g/kg diet) in a control diet was replaced by sorbose, the blood glucose concentration was dramatically lower, but the serum insulin concentrations did not differ. When mice were fed the diets before the onset of diabetic symptoms, glucose excretion in urine was prevented in the mice fed the sorbose diet, but mice fed the control diet excreted glucose in the urine, and the concentration increased with age. When dietary treatment began after the development of diabetic symptoms, dietary sorbose greatly reduced the incidence of hyperglycemia and lowered urinary glucose excretion, compared with mice fed the sucrose-containing diet. These results suggest that dietary sorbose might be useful in patients with, or at risk of developing, noninsulin dependent diabetes, both before and after exhibiting the syndrome. PMID- 8421232 TI - Linolenic acid transport in hamster intestinal cells is carrier-mediated. AB - The intestinal uptake of [1-14C]linolenic acid [18:3(n-3)], an essential fatty acid, was investigated in isolated hamster intestinal cells using a rapid filtration method and 20 mmol/L taurocholate as solubilizing agent. Under these conditions, the initial rate of alpha-linolenic acid uptake was not a linear function of external monomer concentrations in the range of 2 to 2250 nmol/L, but rather the transport system was characterized by saturation kinetics with Vmax = 11.37 nmol.mg protein-1.min-1 and Km = 382 nmol/L. Temperature and metabolic poisons (2,4-dinitrophenol, antimycin A) drastically decreased the initial rate of uptake, as did replacement of Na+. The presence of excess unlabeled alpha linolenic acid in the incubation medium significantly inhibited the uptake of [1 14C]linolenic acid, whereas L-alanine and D-glucose had no effect. Other long chain fatty acids (saturated or unsaturated), as well as cholesterol, inhibited the uptake of [1-14C]linolenic acid. We concluded that an active, carrier mediated mechanism was involved in the intestinal transport of alpha-linolenic acid. Inhibition data are compatible with the hypothesis that intestinal uptake of alpha-linolenic acid is mediated by a carrier common to long-chain fatty acids. PMID- 8421233 TI - Litter size, adrenalectomy and high fat diet alter hypothalamic monoamines in genetically lean (Fa/Fa) Zucker rats. AB - To determine if diet-induced obesity is associated with depressed serotonergic activity (as is genetic obesity), we examined hypothalamic biogenic amines in 11 wk-old genetically lean (Fa/Fa) male Zucker rats raised in small (3 pups/dam) or control (8-9 pups/dam) litters. Five-week-old rats were adrenalectomized or sham operated and, 1 wk later, fed either 11% of energy as fat (low fat) or 68% of energy as fat (high fat) diets for 5 wk. Tissue punches from the ventromedial nucleus (VMN), the paraventricular nucleus and the preoptic area were assayed via HPLC. Rats fed high vs. low fat had a greater percentage of body fat and brown fat mitochondrial GDP binding, whereas serotonergic turnover was lower. Small litter vs. control litter animals had lower VMN and preoptic concentrations of 3,4-dihydroxyphenylacetic acid, a major metabolite of dopamine. Although adrenalectomy resulted in smaller, leaner rats, it did not differentially affect the rats that became fatter. Because VMN and preoptic dopaminergic activities were depressed in small litter vs. control litter rats but the percentage of body fat was unchanged, this decreased dopamine metabolism is probably not causal to the obesity development. However, the same cannot be said for the attenuated serotonergic activity, although such activity may not be directly related to the degree of obesity. PMID- 8421234 TI - Pyridoxine-5'-beta-D-glucoside competitively inhibits uptake of vitamin B-6 into isolated rat liver cells. AB - Uptake of pyridoxine-5'-beta-D-glucoside by freshly isolated rat liver cells was studied at a concentration (0.5 mumol/L) approximating the physiological range of vitamin B-6 by using a membrane filtration method. Unlabeled pyridoxine glucoside was found to competitively inhibit the uptake of [4'-3H]pyridoxine but have no detectable effect on the uptake of D-[1-3H]glucose by hepatocytes. The uptake of [3H]pyridoxine glucoside by isolated rat liver cells was very similar to the uptake of [3H]pyridoxine with Kt = 13.8 mumol/L (6.3 mumol/L for pyridoxine) and Vmax = 82 pmol/(10(6) cells.min) [28 pmol/(10(6) cells.min for pyridoxine)]. The results of this study indicate that pyridoxine glucoside uses the same transport system as does pyridoxine. Upon entry to the cell, pyridoxine glucoside undergoes hydrolysis to release pyridoxine, which is the rate-limiting step in metabolism of this beta-glucoside. PMID- 8421235 TI - Energy restriction reduces metabolic rate in adult male Fisher-344 rats. AB - Energy restriction, without malnutrition, prolongs the maximum life span of laboratory rodents. A reduction in metabolic rate has been proposed as a potential mechanism for increased longevity. The present study examines changes in metabolic rate of adult rats after a 6-wk period of energy restriction. Two groups (n = 6) of 6-mo-old male Fisher-344 rats were studied. Restricted rats were pair-fed a diet equivalent in vitamins and minerals but restricted to 60% of energy consumed by rats eating ad libitum. Average and basal metabolic rates were measured by direct calorimetry over a 24-h period without food. Fat mass and lean body mass were determined by NMR spectroscopy. After 6 wk of restriction, when expressed per kilogram of lean body mass the average metabolic rate was reduced by 14% and basal metabolic rate by 12% compared with the ad libitum diet rats (P < or = 0.01). Reduction of metabolic rate did not seem to be a transient effect of chronic energy restriction in mature rats. PMID- 8421236 TI - Calcium deficiency and food deprivation improve the response of chickens to acute heat stress. AB - The tolerance of chickens to acute heat stress may be modified by diet. Broiler chickens fed calcium-adequate (0.90% Ca) or -deficient (0.45% or 0.15% Ca) diets were either fed or not fed for 24 h and exposed to increasing temperatures (from 24 to 41 degrees C). Diets were fed for 7 d before heat stress in Experiment 1 and for 14 d before heat stress in Experiment 2. Body temperature, blood ionized Ca, pH, pCO2, plasma inorganic phosphate and total Ca were determined. During heat stress, Ca+2 and inorganic phosphate were depressed in all treatments. Feeding the 0.45% Ca diet for 7 d reduced hyperthermic body temperature of fed chickens but had no effect on body temperature of unfed chickens relative to the groups fed 0.90% Ca. No further improvement in body temperature response to heat stress was obtained by lowering the dietary Ca level to 0.15% or extending the feeding period to 14 d. Food deprivation was more effective in counteracting the heat-induced rise in body temperature than a dietary Ca deficiency. Heat-induced changes in body temperature, Ca+2, inorganic phosphate and blood pH were highly correlated (P < 0.001). The change in Ca+2 followed a pattern similar to that of changes in body temperature, but changes in inorganic phosphate seemed to be more indicative of changes in pH. Control birds fed 0.90% Ca exhibited the highest changes in Ca+2 and body temperature values. Feeding Ca-deficient diets reduced changes in both Ca+2 and body temperature. Unfed birds, regardless of dietary Ca level, showed the lowest changes in Ca+2 and body temperature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421237 TI - Discipline: what do you recommend? PMID- 8421238 TI - The parasomnias of childhood. AB - Parents are rarely questioned in detail about the sleep characteristics of their children except during the infancy period. Somnambulism, commonly known as sleepwalking, affects approximately 15% of young children. Safety is the biggest concern for a family whose child sleepwalks. Other parasomnias include night terrors, nightmares, and sleeptalking. This article explores the incidence and characteristics of parasomnias, reviews the physiology of sleep, and offers pediatric nurse practitioners information for history taking and anticipatory guidance. PMID- 8421239 TI - Autism in young children: an update. AB - Autism is a pervasive developmental disorder with onset in infancy or childhood. Research has identified infant/toddler characteristics that should suggest autism to pediatric primary care providers. Many autistic children can be referred for diagnosis as young toddlers if social, perceptual, and language delays are considered significant. Early diagnosis of children with autism allows intervention, which helps the child modulate disturbing sensory stimuli and seeks to meet his/her unique social and communication needs. This article describes the disorder and its management and offers guidelines for recognizing young children who would benefit from further comprehensive neurobiologic evaluations by developmental specialists. PMID- 8421240 TI - Pediatric nurse practitioners specializing with survivors of childhood cancer. AB - The evolution of the role of the pediatric nurse practitioner in oncology specializing in the care of childhood cancer survivors is described, with certain aspects of the role solidified or expanded and new functions added. The early concept of the role included three interdependent functions: (a) clinician/caregiver, (b) educator, and (c) researcher. The functions of specialty care provider and educator remain strong; the role of researcher has been expanded, and new role components, clinical/program manager and consultant, have been added. The central focus for the pediatric nurse practitioner in oncology is the survivor and family, which is extended to the clinic population and related groups by the blending of the pediatric nurse practitioner and clinical nurse specialist roles. Any role function on behalf of this clinical population should be assumed as necessary to provide comprehensive care. PMID- 8421241 TI - AIDS risk among black male adolescents: implications for nursing interventions. AB - Acquired immunodeficiency syndrome (AIDS) is a devastating disease the incidence of which is increasing in the black heterosexual community. Only 12% of the nation's population is black, yet 28% of the people with AIDS are black. AIDS currently threatens black adolescents because of their high risk sexual behavior. No cure exists for AIDS nor is a vaccine available. However, AIDS is preventable. Presently, teaching people how to avoid becoming infected with the human immunodeficiency virus is the only strategy for reducing the spread of AIDS. Nurses can be an important link in the chain of AIDS prevention in this country. Minimal information is available to assist nurses in learning how to effectively intervene with adolescents, especially black male adolescents, regarding AIDS prevention. This article reviews research on AIDS knowledge, attitudes, and high risk sexual behavior among black male adolescents, suggests strategies for using the nursing process as an AIDS education model in working with black male adolescents, and provides guidelines for nurses to use when working with black male adolescents concerning the issue of sexuality education and AIDS prevention. PMID- 8421242 TI - Partners in caring: a state-funded primary care initiative for children. AB - Improving access to health care for children is a growing concern. Health goals for the year 2000, both in the nation and in the state of Wisconsin, underscore the urgency of preventive care. The Partners in Caring program is a state-funded primary care initiative involving 13 suburban public health departments that provide preventive care to uninsured and underserved children in the participating communities of the consortium. This article describes the program during the first year of well child assessments, the role of the pediatric nurse practitioner, the impact of the program, and implications for nursing practice. PMID- 8421243 TI - Hands and feet that blister and peel: dyshidrosis. PMID- 8421244 TI - Gangs. PMID- 8421245 TI - Urinary frequency in an adolescent female. PMID- 8421246 TI - It's the 103rd Congress: do you know how your members of Congress are going to vote? PMID- 8421247 TI - NAPNAP President's message on access to care. PMID- 8421248 TI - Eysenck's arousal theory of introversion-extraversion: a converging measures investigation. AB - Convergent electroencephalographic, performance, and psychometric measures of arousal were used to test predictions based on H. J. Eysenck's (1967) theory. Twelve introverts and 12 extraverts had their brainstem auditory evoked responses (BAERs) monitored under 3 levels of caffeine administration (0.0, 1.5, and 3.0 mg/kg) and 3 levels of task demand (resting, simple response time, and complex response time). Results revealed a decrease in the latency of Wave V of the BAER as a function of personality, with introverts evidencing significantly shorter latency of Wave V as compared with extraverts. Faster conduction time between Waves I-III and I-V were also found in introverts. Caffeine at either the 1.5 or 3.0 mg/kg level was associated with decreased latency of Wave V, compared with the placebo condition. No personality differences in subjective arousal or reaction time performance were found across any of the conditions. PMID- 8421249 TI - Negative life events, perceived stress, negative affect, and susceptibility to the common cold. AB - After completing questionnaires assessing stressful life events, perceived stress, and negative affect, 394 healthy Ss were intentionally exposed to a common cold virus, quarantined, and monitored for the development of biologically verified++ clinical illness. Consistent with the hypothesis that psychological stress increases susceptibility to infectious agents, higher scores on each of the 3 stress scales were associated with greater risk of developing a cold. However, the relation between stressful life events and illness was mediated by a different biologic process than were relations between perceived stress and illness and negative affect and illness. That these scales have independently relations with illness and that these relations are mediated by different processes challenges the assumption that perceptions of stress and negative affect are necessary for stressful life events to influence disease risk. PMID- 8421250 TI - When ego threats lead to self-regulation failure: negative consequences of high self-esteem. AB - The tendency for people with high self-esteem to make inflated assessments and predictions about themselves carries the risk of making commitments that exceed capabilities, thus leading to failure. Ss chose their performance contingencies in a framework where larger rewards were linked to a greater risk of failure. In the absence of ego threat, Ss with high self-esteem showed superior self regulation: They set appropriate goals and performed effectively. Ego threat, however, caused Ss with high self-esteem to set inappropriate, risky goals that were beyond their performance capabilities so they ended up with smaller rewards than Ss with low self-esteem. The results indicate the danger of letting egotistical illusions interfere with self-regulation processes. PMID- 8421251 TI - When does introspection bear fruit? Self-reflection, self-insight, and interpersonal choices. AB - Whereas earlier research suggests that the fruits of introspection may promote error and misperception, this research suggests that thinking about the self may sometimes foster self-insight. Participants who had opportunity to reflect on themselves were particularly inclined to display self-insight by (a) rating feedback that confirmed their self-views as self-descriptive (Experiments 1 and 3), (b) rating themselves in ways that matched their friends' appraisals of them (Experiment 2), and (c) choosing a self-verifying interaction partner rather than an overly favorable one (Experiment 4). These effects were moderated by the nature of the introspective activity (Experiment 3) and by its duration (Experiment 4). Implications of these findings for the nature of self-knowledge and the worlds people construct around themselves are discussed. PMID- 8421252 TI - The stigma of overweight: affective consequences of attributional ambiguity. AB - Attributing negative outcomes to prejudice and discrimination may protect the mood and self-esteem of some stigmatized groups. Thus, the overweight may be low in self-esteem because they blame their weight, but not the attitudes of others, for negative outcomes based on their weight. In an experiment, 27 overweight and 31 normal weight college women received either positive or negative social feedback from a male evaluator. Relative to other groups, overweight women who received negative feedback attributed the feedback to their weight but did not blame the evaluator for his reaction. This attributional pattern resulted in more negative mood for these overweight women in comparison with other groups. Dimensions of stigma that may account for differences in the tendency to attribute negative outcomes to prejudice, and implications of these findings for weight loss programs and psychotherapy for the overweight, are discussed. PMID- 8421253 TI - Behavioral markers and recognizability of the smile of enjoyment. AB - Ekman and Friesen (1982) predicted that smiles that express enjoyment would be marked by smoother zygomatic major actions of more consistent duration than the zygomatic major actions of nonenjoyment smiles. Study 1 measured the duration and smoothness of smiles shown by female subjects in response to positive emotion films while alone and in a social interaction. Enjoyment smiles in both situations were of more consistent duration and smoother than nonenjoyment smiles. In Study 2 observers who were shown videotapes of enjoyment and nonenjoyment smiles were able to accurately identify enjoyment smiles at rates greater than chance; moreover, accuracy was positively related to increased salience of orbicularis oculi action. In Study 3, another group of observers were asked to record their impressions of the smiling women shown in Study 2. These women were seen as more positive when they showed enjoyment compared with nonenjoyment smiles. These results provide further evidence that enjoyment smiles are entities distinct from smiles in general. PMID- 8421254 TI - Social cognitions as organizers of autonomic and affective responses to social challenge. AB - Autonomic and affective responses to children were assessed as a function of adult perceptions of interpersonal control. Women (N = 160) interacted with and provided feedback to computer-simulated children who "behaved" responsively or unresponsively on a computer game. Women were categorized as low in perceived control (PC) if they attributed high control to children but low control to self over negative events on the Parent Attribution Test. As predicted, low-PC women were maximally reactive to child characteristics, manifesting peak levels of defensive arousal (increased level of heart rate and electrodermal activity) and negative affect with unresponsive children and minimal levels of arousal and negative affect with responsive children. Intermediate response levels were shown by high-PC Ss. We interpreted results as suggesting mediating factors that may operate in dysfunctional interaction patterns previously found for low-PC caregivers. PMID- 8421255 TI - Psychosocial stress, cognitive performance and disability after common whiplash. AB - The relationship between psychosocial stress, cognitive performance and disability was assessed in 97 randomly selected common whiplash patients. Patients were investigated early after injury (mean 7.2 days, SD = 3.8) and again at 6 months. Assessment included different aspects of psychosocial stress, negative affectivity, personality traits and attentional functioning. At 6 months six patients (7%) showed partial or complete disability (disabled group) while 91 patients went back to work at pre-injury levels (non-disabled group). However, 26 patients from the latter group at 6 months were still symptomatic. The disabled and non-disabled groups did not differ with respect to psychosocial stress, negative affectivity and personality traits as assessed at baseline. At 6 months no significant differences were found between the disabled group and 26 symptomatic patients from the non-disabled group with respect to any of the assessed factors. The disabled group showed a combination of the following variables as assessed at baseline: greater age, initial neck pain intensity, initial back pain, blurred vision, and anxiety but less dizziness, sensitivity to noise and neurotic or behavioural problems in childhood. PMID- 8421256 TI - Components of Type A behavior and the first-year prognosis of a myocardial infarction. AB - This study examined prospectively the role of Type A behavior and its subcomponents in the first-year prognosis of myocardial infarction (MI). Anger expression, irritability, and cynicism were assessed as traits related to the hostility component of the construct. The sample comprised 92 patients, less than 65 yr old, who survived the acute phase of their first MI. Psychological data was collected by self-report questionnaires during the initial hospital stay. Type A behavior was measured by the Jenkins Activity Survey and by the Finnish Type A Scale. Factors were controlled for age, sex, social status, and the MI severity. The results showed that patient who had a reinfarction or died during the first year reported more irritability and experienced anger more often than patients surviving without any complications. From the standard subcomponents only the Speed-Impatience factor of the JAS predicted poor prognosis. Our results indicated that the global Type A scores were not associated with the prognosis of MI. PMID- 8421257 TI - Development of the Perceived Stress Questionnaire: a new tool for psychosomatic research. AB - A 30-question Perceived Stress Questionnaire (PSQ) was validated, in Italian and English, among 230 subjects. Test-retest reliability was 0.82 for the General (past year or two) PSQ, while monthly Recent (past month) PSQs varied by a mean factor of 1.9 over 6 months; coefficient alpha > 0.9. General and/or Recent PSQ scores were associated with trait anxiety (r = 0.75), Cohen's Perceived Stress Scale (r = 0.73), depression (r = 0.56), self-rated stress (r = 0.56), and stressful life events (p < 0.05). The General PSQ was higher in in-patients than in out-patients (p < 0.05); both forms were correlated with a somatic complaints scale in a non-patient population (r > 0.5), and were higher, among 27 asymptomatic ulcerative colitis patients, in the seven who had rectal inflammation than in those with normal proctoscopy (p = 0.03). Factor analysis yielded seven factors, of which those reflecting interpersonal conflict and tension were significantly associated with health outcomes. The Perceived Stress Questionnaire may be a valuable addition to the armamentarium of psychosomatic researchers. PMID- 8421258 TI - Longitudinal measurement in the diagnostics of the premenstrual syndrome. AB - In this study 51 women, who had stated that they suffered from premenstrual syndrome (PMS), answered a Dutch adaptation of the Moos' Menstrual Distress Questionnaire (MDQ) every other day for at least two consecutive menstrual cycles. Results showed a clear increase in symptomatology starting about seven days before menstruation and reaching a maximum at the second day of the following cycle. Diagnostic criteria to categorize individual women showed, over 2 years, a substantial decrease of the number of women with PMS in one or both of two menstrual cycles. PMID- 8421260 TI - Dimensions of illness behaviour as measured by the Illness Behaviour Questionnaire: a replication study. AB - A principal component analysis was carried out on the 62-item Illness Behaviour Questionnaire responses of 832 pain clinics and patients and 746 psychiatric in patients in the same general hospital. Analyses were carried out for the groups separately, and combined (N = 1578). The factors obtained were compared to those of the original study, Pilowsky and Spence J Psychosom Res 1975, 19: 279-287, and studies by Zonderman et al. Hlth Psychol 1985; 4: 425-436, and Main and Waddell Pain 1987; 28: 13-26. Varying degrees of robustness of the original factors was demonstrated and this is discussed in relation to the clinical context in which the IBQ is used. The significance of five new factors which emerged is discussed. They were labelled 'Interpersonal Sensitivity', 'Responsiveness to Medical Reassurance', 'Illness Impact', 'Self Appraisal' and 'Biological Functions'. PMID- 8421259 TI - Psychosocial factors and clinical evolution in HIV-1 infection: a longitudinal study. AB - The aim of the study was to assess the relationship between the initial psychosocial situation and the probability of later symptom development in HIV-1 infection. One hundred HIV-1 seropositive subjects, 79 in Stage III (LAS) and 21 in Stage II (asymptomatic), were examined both immunologically (CD4+, Skin Test) and psychologically (test battery). Follow-up at 6 and 12 months involved clinical and immunological reassessment of subjects, who were then classified as fully symptomatic (S, Stage IV) or unchanged (U). The two groups were compared through ANOVA on initial psychosocial measures, while stepwise logistic multiple regression was employed to assess the predictive value of psychosocial measures on clinical and immunological evolution. Psychosocial measures most clearly showing an association with clinical evolution were Denial/Repression attitudes (negatively) and Fighting Spirit (positively), whereas aspects of Hardiness and Social Support showed an effect in interaction with initial CD4+ levels. No stable results were obtained on immunologic evolution. The two groups (U and S) did not show significant differences on other independent variables, with the exception of age. PMID- 8421261 TI - The effect of voluntary breathing on reaction time. AB - The effect of voluntary changes in inspiratory duration on reaction time was studied in 17 normal subjects. All received a visual feedback on inspiratory duration and successively adjusted this variable to two target values which were respectively higher and slightly lower than resting level. The order of the two tasks was counterbalanced across subjects. Reaction times to auditory stimuli were measured during a key-pressing task immediately after voluntary breathing. Voluntary control of inspiration induced concomitant changes in breathing period and minute ventilation. Per cent changes in these breathing variables and in reaction times under the two conditions were analysed. Correlations were significant for period not for minute ventilation. This effect was probably due to neuromuscular rather than humoral factors. PMID- 8421262 TI - Effectiveness of self-management programmes and relaxation training in the treatment of bronchial asthma: relationships with trait anxiety and emotional attack triggers. AB - Twenty-seven children with light to moderate asthma were allocated to three groups matched with respect to a number of subjects with (a) high trait anxiety and (b) emotionally triggered attacks (ET). The first group received an asthma self-management programme (SM) and the second the same programme including relaxation training (SM + R); the control group (CG) received only pharmacological treatment. Before and immediately after intervention, and at 6 and 12 month follow-ups, we evaluated a series of clinical and pulmonary function indicators of disease severity. Overall, patients receiving SM showed a significantly greater reduction in their subjective assessment of attack intensity and in the severity of the intervention measures employed. Inclusion of individual difference variables in the analysis also indicated that ET patients receiving SM + R showed significant decreases in attack duration, and improvements in peak expiratory flow rate (PEFR) with respect to ET patients in all other groups. Trait anxiety level did not affect response to either intervention. PMID- 8421263 TI - Transitional housing for veterans with a psychiatric diagnosis. AB - 1. Veterans with a psychiatric diagnosis needed affordable housing and emotional support to successfully reintegrate into the community. 2. A supervising nurse established a structured, transitional housing program with a housing manager and mandatory weekly meetings. 3. The focus of the house program was centered on resident independence and responsibility. 4. The housing program is cost effective and has shown a 79% success rate in assisting clients to become productive members of the community. PMID- 8421264 TI - Dissociation & multiple personality disorder. A challenge for psychosocial nurses. AB - 1. Multiple personality disorder (MPD) is a dissociative disorder in which two or more distinct personality states exist within an individual. At least two of these personalities recurrently take full control of the person's behavior. 2. Most MPD clients report histories of severe childhood trauma, particularly physical, sexual, and ritual cult abuse. MPD most likely originates in childhood, but is not usually diagnosed until adulthood. In most reported cases, the first dissociative episode is thought to have occurred at a very young age. 3. Signs and symptoms that may suggest MPD include a history of medical and psychiatric diagnoses; inconsistencies in accounts of elapsed time and physical behaviors; psychophysiological complaints; experiencing voices inside the head; and an individual referring to herself as "we" instead of "I." PMID- 8421265 TI - Grieving: facilitating the process for dementia caregivers. AB - 1. Grief is a psychological and physiological process that occurs in response to a specific loss. Through the grieving process, those who have experienced a loss come to terms with the finality of the loss and the meaning of the loss in their life. 2. Dementias bring a number of predeath losses that cause caregivers to grieve. Because such losses are often hard to recognize, nurses can plan interventions to facilitate predeath grief. 3. Even when dementia caregivers are able to work through their predeath grief, they cannot fully re-establish their lives until after their relative dies. Some caregivers may never completely resolve the predeath and postdeath losses that have resulted from caring for their relative. PMID- 8421266 TI - A clinical ladder for milieu counselors. An opportunity to contribute to self esteem. AB - 1. Self-esteem affects motivation, achievement, and job satisfaction. 2. Access to a clinical ladder for milieu counselors rewards their achievements in a visible and tangible way, thereby increasing their self-esteem with positive effects on their job satisfaction and quality of patient care. 3. Since the implementation of the clinical ladder program for milieu counselors, there has been documented evidence of increased retention and higher levels of job performance on work evaluations. PMID- 8421267 TI - The problem of missed psychiatric diagnoses in the elderly. PMID- 8421268 TI - Fighting the stigma. PMID- 8421269 TI - Music therapy credentials. PMID- 8421270 TI - The voodoo religion. PMID- 8421271 TI - A view from both sides. PMID- 8421272 TI - Come out! PMID- 8421273 TI - Contracting to lower precaution status for child psychiatric inpatients. AB - 1. Contracting will reduce high-risk precaution incidents. 2. Children are capable of understanding, writing, and adhering to contracting procedures. 3. Contracting performed according to established procedures that use specified criteria for advancement is effective. 4. Nurses should listen carefully and critically examine the child's statements, avoiding making inferences about the child's intentions. PMID- 8421275 TI - Oxidation chemistry of the endogenous central nervous system alkaloid salsolinol 1-carboxylic acid. AB - The oxidation chemistry of salsolinol-1-carboxylic acid (1), an alkaloid endogenous to the central nervous system which is elevated as a result of ethanol consumption, has been studied by electrochemical approaches at pH 7.0 in aqueous solution. The first voltammetric oxidation peak of Ia of 1 at pH 7.0 occurs at Ep = +0.116 V, indicating that this alkaloid is a very easily oxidized compound. The peak Ia reaction is a 2e-2H+ oxidation of 1 to 1,2,3,4-tetrahydro-1-methyl-1 carboxy-6,7-isoquinolinedione (8), which rapidly decarboxylates (k > 10(3) s-1) to give predominantly the quinone methide tautomer of 3,4-dihydro-1-methyl-6,7 isoquinolinediol (2). The latter compound is responsible for the second observed oxidation peak IIa observed with 1. This peak is a 2e oxidation of 2 to a quinoid intermediate (9) which can either be attacked by water to yield 3,4-dihydro-1 methyl-5,7-dihydroxyisoquinolin-6-one (13b) (which is readily further oxidized to 3,4-dihydro-1-methyl-5-hydroxyisoquinoline-6,7-dione (3)) or aromatizes to yield 1-methyl-6,7-isoquinolinediol (4). Preliminary in vivo experiments have revealed that 2 and 13b are behavioral toxins when injected into the brains of laboratory mice. The in vitro oxidation reactions of 1 and 2 reported here might be of relevance to the neurodegenerative, behavioral, and addictive consequences of chronic alcoholism. PMID- 8421274 TI - Nonpeptide angiotensin II antagonists: N-phenyl-1H-pyrrole derivatives are angiotensin II receptor antagonists. AB - A series of 5-[1-[4-[(4,5-disubstituted-1H-imidazol-1-yl)methyl]- substituted]-1H pyrrol-2-yl]-1H-tetrazoles and 5-[1-[4-[(3,5-dibutyl-1H-1,2,4-triazol-1 yl)methyl]-substituted]- 1H-pyrrol-2-yl]-1H-tetrazoles were investigated as novel AT1-selective angiotensin II receptor antagonists. Computer-assisted modeling techniques were used to evaluate structural parameters in comparison to the related biphenyl system. New synthetic procedures have been developed to prepare the novel compounds. The best antagonists in this series had IC50 values (rat uterine membrane receptor binding) in the 10(-8) M range and corresponding pA2 in isolated organ assay (rabbit aorta rings). Structure-activity relationships indicate some similarities with the finding in the biphenyl system. Substitution on the pyrrole ring modulates activity. Compound 5 antagonized angiotensin induced blood pressure increase when administered to conscious rat at 30 mg/kg per os. PMID- 8421276 TI - Synthesis and use of the n-bromododecane-1,12-diols as conformational probes for general anesthetic target sites. AB - The n-bromododecane-1,12-diols with bromine on carbons 2, 3, 5, and 6, respectively, were synthesized and found to be potent general anesthetics. They were also found to be potent inhibitors of firefly luciferase, a protein model for the primary target sites underlying general anesthesia. However, their effects on lipid bilayers were small, lowering the chain-melting phase transition temperature by less than 1 degree C at their EC50 concentrations for general anesthesia. A large dependence upon the position of the bromine atom was found for both n-hexadecane/water partition coefficients and inhibition constants for firefly luciferase; a much smaller positional dependence was found for induction of general anesthesia and for disrupting lipids. These results are consistent with the bulky bromine atom inhibiting the conformational flexibility of the diol hydrocarbon chain, making these bromo diols useful probes for ascertaining the shapes of apolar binding sites. In particular, our measurements suggest that these novel anesthetics produce general anesthesia by binding to long and relatively narrow apolar target sites in the central nervous system. PMID- 8421277 TI - Purine 2'-deoxy-2'-fluororibosides as antiinfluenza virus agents. AB - Twenty purine 2'-deoxy-2'-fluororibosides were synthesized by enzymic pentosyl transfer from 2'-deoxy-2'-fluorouridine. Each nucleoside analogue was assayed for cytotoxicity in uninfected Madin-Darby canine kidney cells and for their ability to suppress influenza A virus infections in these cells. The most potent antivirial activity was observed with analogues having an amino group in the 2 position of the purine moiety. All 2-unsubstituted analogues were less potent than their 2-amino counterparts. Furthermore, 2-methyl,2-methoxy, or 2-fluoro substitution obliterated antivirial activity. The most cytotoxic member of the series was the 2-fluoro-6-amino analogue (IC50 = 120 microM). 2'-Deoxy-2' fluoroguanosine and those congeners readily converted to it by adenosine deaminase showed the most potent antivirial activity (IC50 = 15-23 microM). Little cytotoxicity was observed with this subgroup of analogues which renders them worthy of further investigation as potential antiinfluenza agents. PMID- 8421278 TI - Use of affinity capillary electrophoresis to determine kinetic and equilibrium constants for binding of arylsulfonamides to bovine carbonic anhydrase. AB - Affinity capillary electrophoresis (ACE) provides a new approach to studying protein-ligand interactions. The basis for ACE is the change in the electrophoretic mobility of the protein when it forms a complex with its ligand. This binding interaction can be quantified directly for charged ligands or indirectly for neutral ligands in competition with a previously characterized charged ligand. Determination of kinetic and equilibrium constants using ACE relies only on the changes in the migration time and shape (but not the area) of the peak due to protein. Simulation of the protein mobility under conditions of ACE suggests that the experimentally obtained electropherograms can be explained in terms of few variables: on and off rates (and thus, binding constant), concentration of the ligand(s), and relative mobilities of the protein and its complex(es). PMID- 8421279 TI - Pyrazoline bisphosphonate esters as novel antiinflammatory and antiarthritic agents. AB - Vinylidenebisphosphonic acid tetraethyl ester (1) and diazo ketones 7a-1 in ether at 22 degrees C yield pyrazoline bisphosphonate tetraethyl esters 8a-1 in moderate to good yield. These compounds were evaluated in animal models of arthritis: rat adjuvant-induced polyarthritis (AIP) and murine antigen-induced arthritis (AIA) and a murine model of chronic inflammation, the delayed type hypersensitivity granuloma reaction (DTH-GRA). (5-Benzoyl-2,4-dihydro-3H-pyrazol 3-ylidene)-bisphosphonic acid tetraethyl ester (8a), and [5-(3-fluorobenzoyl)-2,4 dihydro-3H-pyraxol-3-ylidene]- bisphosphonic acid tetraethyl ester (8d) significantly inhibited the arthritis models, AIP (15 mg/kg) and AIA (25 mg/kg), as well as the DTH-GRA (25 mg/kg). Conversion of 8a to the corresponding bisphosphonic acid, 10a, resulted in loss of activity. Compounds with alkyl substituents on the pyrazoline nitrogen, 9a-d, were inactive in the DTH-GRA. These results show that 8a and 8d have novel antiinflammatory activity and are capable of inhibiting chronic arthritis and inflammation in animals. Such compounds might be useful in man for treating chronic tissue injury associated with arthropathies such as inflammatory joint disease as well as other chronic inflammatory diseases. PMID- 8421280 TI - Further development of hydrogen bond functions for use in determining energetically favorable binding sites on molecules of known structure. 1. Ligand probe groups with the ability to form two hydrogen bonds. AB - The directional properties of hydrogen bonds play a major role in determining the specificity of intermolecular interactions. An energy function which takes explicit account of these properties has been developed for use in the determination of energetically favorable ligand binding sites on molecules of known structure by the GRID method (Goodford, P.J.J. Med. Chem. 1985, 28, 849. Boobbyer, D.N.A.; Goodford, P.J.; McWhinnie, P.M.; Wade, R.C.J. Med. Chem. 1989, 32, 1083). In this method, the interaction energy between a target molecule and a small chemical group (a probe), which may be part of a larger ligand, was calculated using an energy function consisting of Lennard-Jones, electrostatic, and hydrogen bond terms. The latter term was a function of the length of the hydrogen bond, its orientation at the hydrogen-bonding atoms, and their chemical nature. We now describe hydrogen bond energy functions which take account of the spatial distribution of the hydrogen bonds made by probes with the ability to form two hydrogen bonds. These functions were designed so as to model the experimentally observed angular dependence of the hydrogen bonds. We also describe the procedure to locate the position and orientation of the probe at which the interaction energy is optimized. The use of this procedure is demonstrated by examples of biological and pharmacological interest which show that it can produce results that are consistent with other theoretical approaches and with experimental observations. PMID- 8421281 TI - Further development of hydrogen bond functions for use in determining energetically favorable binding sites on molecules of known structure. 2. Ligand probe groups with the ability to form more than two hydrogen bonds. AB - The specificity of interactions between biological macromolecules and their ligands may be partially attributed to the directional properties of hydrogen bonds. We have now extended the GRID method (Goodford, P. J. J. Med. Chem. 1985, 28, 849. Boobbyer, D. N. A.; Goodford, P. J.; McWhinnie, P. M.; Wade, R. C. J. Med. Chem. 1989, 32, 1083), of determining energetically favorable ligand binding sites on molecules of known structure, in order to improve the treatment of groups which can make multiple hydrogen bonds. In this method, the interaction energy between a probe (a small chemical group that may be part of a larger ligand) and a target molecule is calculated using an energy function which includes a hydrogen bond term which is dependent on the length of the hydrogen bond, its orientation at the hydrogen-bonding atoms, and their chemical character. The methods described in the preceding paper (Wade, R. C.; Clark, K. J.; Goodford, P. J. J. Med. Chem., preceding paper in this issue) for probes capable of making two hydrogen bonds are here extended to the following probes which have the ability to make more than two hydrogen bonds: ammonium-NH3+, amine NH2, sp3-hybridized hydroxyl, and water. Use of the improved GRID procedure is demonstrated by the determination of the conformation of an amino acid side chain at the subunit interface in hemoglobin and of the location of water binding sites in human lysozyme. PMID- 8421282 TI - Synthesis and structure-activity relationship of some 5-[[[(dialkylamino)alkyl]-1 piperidinyl]acetyl]-10,11-dihydro-5H- benzo[b,e][1,4]diazepin-11-ones as M2 selective antimuscarinics. AB - A series of 5-[[[(dialkylamino)alkyl]-1-piperidinyl]acetyl]- 10,11-dihydro-5H dibenzo[b,e][1,4]-diazepin-11-ones were prepared as potential M2-selective ligands. The compounds were evaluated for their affinity and selectivity for the muscarinic cholinergic receptor. The best M2-selective antimuscarinic agent studied is 5-[[4-[4-diethylamino)butyl]-1- piperidinyl]acetyl]-10,11-dihydro-5H dibenzo[b,e][1,4]diazepin-11- one, which is approximately 10 times more potent at M2 receptors than previously known compounds such as 11-[[4-[4 (diethylamino)butyl]- 1-piperidinyl]acetyl]-5,11-dihydro-6H- pyrido[2,3 b][1,4]benzodiazepin-6-one (AQ-RA 741). PMID- 8421283 TI - CCK-B agonist or antagonist activities of structurally hindered and peptidase resistant Boc-CCK4 derivatives. AB - Replacement of Met31 by (N-Me)Nle in CCK8 or CCK4 has been shown to improve the affinity and selectivity for CCK-B receptors. In order to obtain molecules with enhanced bioavailability, two novel series of protected tetrapeptides of the general formula Boc-Trp30-X-Asp-Y33 have been developed. Introduction of (N Me)Nle and the bulky, aromatic naphthylalaninamide (Nal-NH2) in positions X and Y, respectively, does not greatly modify the affinity for guinea pig brain CCK-B receptors. In contrast, incorporation of hindering N-methyl amino acids such as (N-Me)Phe, (N-Me)Phg, or (N-Me)Chg, but not their non-methylated counterparts, in position X induced a large decrease in affinity for the CCK-B binding sites. Among the various peptides synthesized, Boc-[(N-Me)Nle31,1Nal-NH2(33)]CCK4 (2) (KI = 2.8 nM), Boc-[Phg31,1Nal-NH2(33)]CCK4 (15) (KI = 14 nM), and Boc [Phg31,1Nal-N(CH3)2(33)]CCK4 (17) (KI = 39 nM) displayed good affinities for brain CCK-B receptors and had good selectivity ratios. These pseudopeptides, in which the presence of unnatural and hydrophobic residues is expected to improve their penetration of the central nervous system, were shown to be very resistant to brain peptidases. Interestingly, whereas compounds 2 and 15 proved to be full agonists for rat hippocampal CCK-B receptors when measured in an electrophysiological assay, compound 17 behaved as a potent antagonist in the same test and displayed a good affinity in rat brain KI(CCK-B) = 51 nM as compared to the Merck antagonist L365,260,KI(CCK-B) = 12 nM. This illustrates a simple means to obtain CCK-B antagonists and suggests that the free, CONH2 group plays a critical role in the recognition of the agonist state of brain CCK-B receptors. PMID- 8421284 TI - Phosphorus-containing inhibitors of endothelin converting enzyme: effects of the electronic nature of phosphorus on inhibitor potency. PMID- 8421285 TI - Hemicholinium and related lipids: inhibitors of protein kinase C. PMID- 8421286 TI - Inhibitors of protein kinase C. 3. Potent and highly selective bisindolylmaleimides by conformational restriction. AB - The protein kinase inhibitor staurosporine has been used to design a series of selective bisindolylmaleimide inhibitors of protein kinase C (PKC). Guided by molecular graphics, conformational restriction of the cationic side chain has led to ATP competitive inhibitors of improved potency and selectivity. Two compounds have been further evaluated and were shown to inhibit PKC of human origin and prevent T-cell activation in a human allogeneic mixed lymphocyte reaction. One of these compounds was orally absorbed in mice and antagonized a phorbol ester induced paw edema in a dose-dependent manner. This compound also selectively inhibited the secondary T-cell mediated response in a developing adjuvant arthritis model in rats and provides evidence for the potential use of PKC inhibitors as therapeutic immunomodulators. PMID- 8421287 TI - 1,3-dioxolanylpurine nucleosides (2R,4R) and (2R,4S) with selective anti-HIV-1 activity in human lymphocytes. AB - In order to study the structure-activity relationships of dioxolane nucleosides as potential anti-HIV-1 agents, various enantiomers of pure dioxolanylpurine nucleosides were synthesized and evaluated against HIV-1 in human peripheral blood mononuclear cells. The enantiomerically pure key intermediate 1, which was synthesized in nine steps from 1,6-anhydro-beta-D-mannose, was condensed with 6 chloropurine, 6-chloro-2-fluoropurine, and 2,6-dichloropurine in the presence of TMS triflate. The chloro or fluoro substituents were readily converted into amino, N-methylamino, hydroxy, methoxy, thiol, and methylthio under appropriate reaction conditions. Upon evaluation of these dioxolanes, the guanine derivative 24 exhibited the most potent anti-HIV-1 activity without cytotoxicity up to 100 microM in various cells. The decreasing antiviral activity order of beta-isomers was as follows: guanine > 6-chloro-2-aminopurine > 2-fluoroadenine > or = adenine > or = 2,6-diaminopurine > hypoxanthine > 2-chloroadenine > 6-chloropurine approximately equal to N6-methyladenine approximately equal to 6-mercaptopurine approximately equal to 6-(methylthio)purine. PMID- 8421288 TI - 6-[(aminoalkyl)amino]-substituted 7H-benzo[e]perimidin-7-ones as novel antineoplastic agents. Synthesis and biological evaluation. AB - A class of chromophore-modified anthracenediones with an additional pyrimidine ring incorporated into the chromophore system has been obtained in an attempt to provide compounds with diminished peroxidation activity and thus potentially lowered cardiotoxicity. Their synthesis was carried out by the reaction of 6 amino- or 6-hydroxy-7H-benzo[e]perimidin-7-one with a number of alkylamines. Potent activity was demonstrated in vitro against murine L1210 leukemia cells (equipotent with ametantrone) as well as against P388 leukemia in vivo (% T/C = 130-255). We observed that the benzoperimidines did not stimulate free radical formation, perhaps due to their poor substrate properties for NADH dehydrogenase. PMID- 8421289 TI - Synthesis of 20-hydroxy-, 20-amino-, and 20-nitro-14-hydroxy-21-nor-5 beta,14 beta-pregnane C-3 glycosides and related derivatives: structure-activity relationships of pregnanes that bind to the digitalis receptor. AB - The preparation of derivatives of 14-hydroxy-21-nor-5 beta,14 beta-pregnane and 5 beta,14 beta-pregnane C-3 alpha-L-rhamnosides and tris-beta-D-digitoxosides is described. These derivatives, possessing a C-17 beta COCH2OH, CH2OH, CO2H, CO2Me, CH2NH2, or CH2NO2 group, bind to the digitalis receptor recognition site of heart muscle as measured in a radioligand binding assay. The 21-norpregnane derivatives consistently show greater binding affinity than the corresponding 20 alpha- and 20 beta-pregnane analogs. The C-20 nitro rhamnoside is comparable to digitoxin in binding affinity. The 17 beta-CH2NO2 group is the most effective replacement for the unsaturated lactone in the binding assay found so far, showing binding affinity comparable to that of the cardiac glycosides. PMID- 8421290 TI - S-adenosylmethionine decarboxylase inhibitors: new aryl and heteroaryl analogues of methylglyoxal bis(guanylhydrazone). AB - A series of 3-acylbenzamidine (amidino)hydrazones 7a-h, the corresponding (hetero)aromatic congeners 7i-p, and 3,3'-bis-amidino-biaryls 25a-e were synthesized. The hydrazones 7a-p were prepared by conversion of the corresponding acyl nitriles 1a,c-d,i,n-p to the imido esters 3a,c-d,i and the amidines 5a,c-d,h i, followed by a reaction with aminoguanidine, or vice versa. Similarly, the biaryl 3,3'-dinitriles 23a-e were converted, via the imino esters 24a-c or the imino thioesters 27d-e, to the diamidines 25a-e. These new products are conformationally constrained analogues of methylglyoxal bis(guanylhydrazone) (MGBG). They are up to 100 times more potent as inhibitors of rat liver S adenosylmethionine decarboxylase (SMDC) and generally less potent inhibitors of rat small intestine diamine oxidase (DAO) than MGBG. Some of these SAMDC inhibitors, e.g., compounds 7a, 7e, 7i, 25a, and 25d, have shown antiproliferative effects against T24 human bladder carcinoma cells. These products, whose structure-activity relationships are discussed, are of interest as potential anticancer agents and drugs for the treatment of protozoal and Pneumocystis carinii infections. PMID- 8421291 TI - Structure-based design of inhibitors of purine nucleoside phosphorylase. 1. 9 (arylmethyl) derivatives of 9-deazaguanine. AB - Purine nucleoside phosphorylase (PNP, EC 2.4.2.1) is a salvage enzyme important to the T-cell-mediated part of the immune system and as such is an important therapeutic target. This paper describes the design, synthesis, and enzymatic evaluation of potent, competitive inhibitors of PNP. Potential inhibitors were designed using the three-dimensional structure of the enzyme in an iterative process that involved interactive computer graphics to model the native enzyme and complexes of it with the inhibitors, Monte Carlo-based conformational searching, and energy minimization. Studies of the enzyme/inhibitor complexes were used to determine priorities of the synthetic efforts. The resulting compounds were then evaluated by determination of their IC50 values and by X-ray diffraction analysis using difference Fourier maps. In this manner, we have developed a series of 9-(arylmethyl)-9-deazapurines (2-amino-7-(arylmethyl)-4H pyrrolo[3,2-d]-pyrimidin-4-ones) that are potent, membrane-permeable inhibitors of the enzyme. The IC50 values of these compounds range from 17 to 270 nM (in 1 mM phosphate), with 9-(3,4-dichlorobenzyl)-9-deazaguanine being the most potent inhibitor. X-ray analysis explained the role of the aryl groups and revealed the rearrangement of hydrogen bonds in the binding of the 9-deazaguanines in the active site of PNP relative to the binding of the 8-aminoguanines that results in more potent inhibition of the enzyme. PMID- 8421292 TI - Structure-activity relationship of 99mTc complexes of phenylenediamine-thiol thioether ligands (PhAT) to brain uptake in rats. AB - A novel class of ligands, phenylenediamine-thiol-thioether (PhAT), was synthesized, and their 99mTc complexes were evaluated for potential use as a functional brain imaging agent. The ligands reacted with Na99mTcO4 and SnCl2 to form single, stable, neutral, and lipophilic 99mTc complexes. Several of these complexes showed significant brain uptake and retention in rats. In particular, the S-ethyl, allyl, and propargyl derivatives had high initial brain uptake (0.88, 0.99, and 0.82% dose/g at 5 min, respectively) and good retention (0.71, 0.75, and 0.67% dose/g at 30 min). The structure-activity relationship of alkyl, alkenyl, and alkynyl thioether derivatives is reported. PMID- 8421293 TI - Exploration of neutral endopeptidase active site by a series of new thiol containing inhibitors. AB - With the aim of characterizing the active site of the neutral endopeptidase [EC 3.4.24.11 (NEP)] and especially its putative S1 subsite, two series of new thiol inhibitors designed to interact with the S1, S'1, and S'2 subsites of the enzyme have been synthesized. These molecules correspond to the general formula HSCH(R1)CH(R2)CONHCH(R3)COOH (series I) and HSCH(R1)CH(R2)CONHCH(R3)CONHCH(R4)COOH (series II). Due to the synthetic pathway used, these inhibitors were obtained as mixtures of four stereoisomers. HPLC separation of the stereoisomers of 17 HSCH[CH2CH(CH3)2]CH(CH2Ph)CONHCH(CH3)COOH allowed the stereochemical dependence of the inhibitory potency to be determined. The most active isomer 17b (IC50 = 3.6 nM) is assumed to have the S,S,S stereochemistry, as deduced from both NMR and HPLC data. Although none of the inhibitors obtained were significantly more active than thiorphan, HSCH2CH(CH2Ph)CONHCH2COOH (IC50 = 4 nM), which interacts only with the S'1 and S'2 subsites of NEP, their enhanced hydrophobicity is expected to improve their pharmacokinetic properties. All these compounds displayed low affinities for ACE (IC50s > 1 microM). The determination of the IC50s of two inhibitors of series II for NEP and for a mutated enzyme in which Arg102 was replaced by Glu102 allowed their mode of binding to the active site of NEP to be characterized. The R2 and R3 chains fit the S'1-S'2 subsites, while the R4 group is probably located outside the active site. Taken together these results indicate that the R1 chain of these inhibitors creates no additional stabilizing interactions with the active site of NEP. Two hypotheses may account for this: there is no hydrophobic S1 subsite in NEP or the inhibitors have structures which are too constrained for optimized interactions with the active site. PMID- 8421294 TI - Structural determinants of haloenol lactone-mediated suicide inhibition of canine myocardial calcium-independent phospholipase A2. AB - Haloenol lactones are potent mechanism-based inhibitors of a novel class of calcium-independent phospholipases A2 which have been implicated as the enzymic mediators of membrane dysfunction during myocardial ischemia (Hazen, S. L.; et al. J. Biol. Chem. 1991, 266, 7227-7232). Herein we demonstrate that the ring size, hydrophobic group, and cryptic electrophile in the haloenol lactone moiety are important and modifiable determinants of the inhibitory potency of haloenol lactone-mediated inhibition of calcium-independent phospholipase A2. Direct comparisons between haloenol lactone-mediated inhibition of calcium-independent phospholipase A2 and the absence of inhibition with calcium-dependent phospholipase A2 further underscore the marked differences in the catalytic strategy employed by these two classes of intracellular phospholipases A2. PMID- 8421295 TI - Spacing requirements between LexA operator half-sites can be relaxed by fusing the LexA DNA binding domain with some alternative dimerization domains. AB - The dimerization domain of the LexA repressor has been replaced by two heterologous dimerization motifs: the "leucine zipper" from the jun oncogene product and the carboxy-terminal oligomerization domain of Escherichia coli lac repressor. The corresponding hybrid proteins LexA1-87-Jun zipper and LexA1-87-lac repressor have been purified and their DNA binding properties have been studied using gel mobility shift assays. Both fusion proteins form stable specific complexes with a short DNA duplex harboring the CTGT(at)4ACAG consensus sequence of the LexA repressor. This conserved DNA binding capacity distinguishes these two fusion proteins from many others containing a LexA DNA binding domain fused to different heterologous transactivation and/or dimerization domains. However the fusion proteins LexA1-87-Jun zipper and LexA1-87-lac repressor behave differently from native LexA repressor in that these fusion proteins tolerate the insertion of additional base-pairs between the two invertedly repeated CTGT motifs. LexA1-87-Jun zipper requires two CTGT motifs and tolerates the insertion of at least two additional base-pairs between these motifs, whereas LexA1-87-lac repressor requires in fact only a single CTGT motif for the formation of a specific complex detectable in gel mobility shift assays. The inability of the normal LexA repressor to form well-defined complexes with operators containing additional base-pairs in the center suggests that the LexA "hinge region" between the amino-terminal DNA binding and the carboxy-terminal dimerization domain might not be entirely flexible. In an attempt to remove a hypothetical interaction between the LexA cleavage site (which is situated within the hinge region) and the catalytic cleavage center (situated within the carboxy-terminal domain) a LexA mutant repressor containing five simultaneous mutations in the hinge region has been constructed and purified. Surprisingly this mutant repressor failed to form stable complexes detectable by the gel mobility shift assay even with the normal consensus sequence, suggesting that the LexA hinge region is more than a simple connector between the two structural domains and that its chemical nature is important not only for LexA cleavage, but also for the formation of stable LexA-DNA complexes. PMID- 8421296 TI - Conformational filtering in polypeptides and proteins. AB - We present a method for assigning an ensemble of conformational states to each amino acid residue of a sequence. The states are defined as regions in the (phi, psi) map. The procedure is based on the use of conformational filters. In each filter we use a different set of approximations to estimate the probability of conformational states, and retain only the ones whose probability exceeds an acceptance probability. The resulting state assignment is not necessarily unique, but provides information that can be further exploited in searches for the tertiary structure. This conformational filtering approach to the de novo analysis of a sequence has a number of advantages over traditional structure prediction. First, it is possible to select acceptance probabilities such that the true conformational state is retained for up to 87% of residues, while substantially reducing the number of potential conformations. Second, in solution most linear peptides are present as ensembles of rapidly interconverting conformers, and such ensembles can be well predicted by filtering. Third, we can use Markov chains instead of a statistical mechanical (Ising) treatment, and avoid the need for estimating statistical weight matrices valid for the molecule as a whole. Markov models can use local transition matrices that are assumed to be independent of the rest of the chain, and are directly calculated from pairwise data. We show here that the locally identifiable transition matrices are transferable from the crystal structures of proteins to the solution structures of short peptides, and the ensembles of filtered conformations are in good agreement with nuclear magnetic resonance data. When applied to proteins, the filters retain several conformational states for most residues, and provide a measure of conformational variability. Small variability means that the segment is well defined by local interactions alone, and hence is likely to preserve its structure when isolated from the rest of the chain. Conversely, the structure of a segment with above-average conformational variability is likely to be significantly affected by its protein environment. PMID- 8421297 TI - Molecular shape of vinculin in aqueous solution. AB - We have investigated the molecular structure of chicken gizzard vinculin in solution. The translational diffusion coefficient of the intact protein and its amino-terminal head fragment, as obtained by proteolytic digestion, was determined by photon correlation spectroscopy. The experimental data are compared with hydrodynamic calculations, where the anisotropic shape of the macromolecule is modeled by spherical subunits. Our results are in agreement with the concept of a "balloon on a string" for the molecular shape of native vinculin. The existence of dimer and oligomer structures in low ionic strength buffer can be excluded. The calculated dimensions of the head fragment were estimated to r = 3.3 nm for a spherical particle, but the diffusion coefficient suggests a slightly anisotropic shape. In solution, the rod-like tail exhibits some flexibility, which is probably located in the "neck region" of the protein, considering the known sequence data. PMID- 8421298 TI - Crystal structure of glucose oxidase from Aspergillus niger refined at 2.3 A resolution. AB - Glucose oxidase (beta-D-glucose: oxygen 1-oxidoreductase, EC 1.1.3.4) is an FAD dependent enzyme that catalyzes the oxidation of beta-D-glucose by molecular oxygen. The crystal structure of the partially deglycosylated enzyme from Aspergillus niger has been determined by isomorphous replacement and refined to 2.3 A resolution. The final crystallographic R-value is 18.1% for reflections between 10.0 and 2.3 A resolution. The refined model includes 580 amino acid residues, the FAD cofactor, six N-acetylglucosamine residues, three mannose residues and 152 solvent molecules. The FAD-binding domain is topologically very similar to other FAD-binding proteins. The substrate-binding domain is formed from non-continuous segments of sequence and is characterized by a deep pocket. One side of this pocket is formed by a six-stranded antiparallel beta-sheet with the flavin ring system of FAD located at the bottom of the pocket on the opposite side. Part of the entrance to the active site pocket is at the interface to the second subunit of the dimeric enzyme and is formed by a 20-residue lid, which in addition covers parts of the FAD-binding site. The carbohydrate moiety attached to Asn89 at the tip of this lid forms a link between the subunits of the dimer. PMID- 8421299 TI - Crystal structure of an endochitinase from Hordeum vulgare L. seeds. AB - Higher plants contain multiple constitutively expressed proteins for defense against infection by viruses, bacteria, and fungi. One such class of proteins, the chitinases, are effective antifungal agents because they hydrolyze the insoluble beta-1,4-linked polymer of N-acetylglucosamine (chitin), which is the major component of the mycelial cell wall of many fungi. We report here the three dimensional, 2.8 A, crystal structure of a 26 kDa endochitinase from barley (Hordeum vulgare L.) seeds. The 243 amino acid residue molecule is rich in alpha helices and has three disulfide bonds. A prominent elongated cleft runs the length of the molecule, and is presumably the region responsible for substrate binding and catalysis. Endochitinases from various species of plants show a high degree of similarity in their amino acid sequences. It is therefore likely that the barley endochitinase structure can serve as a model for other plant endochitinases and that catalytic models based on that structure will be applicable to the entire enzyme family. PMID- 8421300 TI - Fragment ranking in modelling of protein structure. Conformationally constrained environmental amino acid substitution tables. AB - Conformationally constrained environment-dependent amino acid residue substitution tables have been constructed from a database comprising 33 homologous families of protein sequences aligned on the basis of their three dimensional structures. Residues are allotted to one of 216 (or 54) classes of combinations of structural features. These include nine main-chain conformation classes, three classes of side-chain accessibility and eight (or two) classes of side-chain involvement in three types of hydrogen bond. Seven different main chain conformational classes outside of regions of regular structure were identified in an analysis of the distributions of phi-psi torsion angles in 84 high-resolution crystallographic structures. Residue substitutions at equivalent positions in the structural alignments are included where the main-chain conformational class is conserved. Frequency data in the form of 216 (or 54) environment specific (20 x 20 residue type) matrices are then converted to probabilities. Two smoothing regimes incorporating entropy-driven weights were applied to the set of 54 tables. Predicted residue substitutions have been generated for individual residue positions in beta-hairpins and the hypervariable regions of the immunoglobulins. These have been compared with the observed sequence variation at the same positions using rank correlation methods. Measurements of chi 2 distances demonstrate the considerable improvement in predictive power at key residue positions identified from interactive graphics studies when compared to the Dayhoff MDM250 mutation matrix. An illustrative example is given of an application of the method in the ranking of loop fragments in model building studies of structurally variable regions in two subtilisins. A combined template scoring procedure is found to be 26-fold more discriminatory than the Dayhoff matrix. The success rate is approximately 85%. PMID- 8421301 TI - Structure of Sesbania mosaic virus at 4.7 A resolution and partial sequence of the coat protein. AB - Sesbania mosaic virus (SMV) is a plant virus infecting Sesbania grandiflora plants in Andhra Pradesh, India. Amino acid sequence of the tryptic peptides of SMV coat protein were determined using a gas phase sequenator. These sequences showed identical amino acids at 69% of the positions when aligned with the corresponding residues of southern bean mosaic virus (SBMV). Crystals diffracting to better than 3 A resolution were obtained by precipitating the virus with ammonium sulphate. The crystals belonged to rhombohedral space group R3 with a = 291.4 A and alpha = 61.9 degrees. Three-dimensional X-ray diffraction data on these crystals were collected to a resolution of 4.7 A, using a Siemens-Nicolet area detector system. Self-rotation function studies revealed the icosahedral symmetry of the virus particles, as well as their precise orientation in the unit cell. Cross-rotation function and modelling studies with SBMV showed that it is a valid starting model for SMV structure determination. Low resolution phases computed using a polyalanine model of SBMV were subjected to refinement and extension by real-space electron density averaging and solvent flattening. The final electron density map revealed a polypeptide fold similar to SBMV. The single disulphide bridge of SBMV coat protein is retained in SMV. Four icosahedrally independent cation binding sites have been tentatively identified. Three of these sites, related by a quasi threefold axis, are also found in SBMV. The fourth site is situated on the quasi threefold axis. Aspartic acid residues, which replace Ile218 of SBMV from the quasi threefold-related subunits are suitable ligands to the cation at this site. PMID- 8421302 TI - Mutational analysis of the function of Gln115 in the EcoRI restriction endonuclease, a critical amino acid for recognition of the inner thymidine residue in the sequence -GAATTC- and for coupling specific DNA binding to catalysis. AB - The Gln115 residue of the EcoRI restriction endonuclease has been proposed to form a hydrophobic contact to the methyl group of the inner thymidine of the EcoRI recognition sequence -GAATTC- and to be involved in intramolecular hydrogen bonds to the mainchain at positions 140 and 143 as well as to the side-chain of Asn173. We have exchanged Gln115 for Ala and Glu by site-directed mutagenesis and analysed the purified mutant proteins (Q115A and Q115E) biochemically and physico chemically. Q115A and Q115E have the same secondary structure composition as wild type EcoRI but are less stable towards thermal denaturation than the wild-type enzyme. In contrast to wild-type EcoRI the mutant proteins show a biphasic denaturation profile under alkaline pH, presumably because the amino acid exchange labilizes one part of the molecule, which unfolds before the rest of the protein is denatured. Q115A is catalytically inactive under normal buffer conditions, in part due to a diminished affinity towards DNA. At low ionic strength and alkaline pH, as well as in the presence of Mn2+, i.e. under conditions where wild-type EcoRI shows a relaxed specificity, Q115A is active, however not as much as wild-type EcoRI. Under these conditions it cleaves the canonical sequence -GAATTC- with the same kcat/Km value as the sequence -GAAUTC-, which differs from the former sequence by a single methyl group, while wild-type EcoRI shows a tenfold lower kcat/Km for cleavage of -GAAUTC- than for -GAATTC-. Binding experiments, carried out in the absence of Mg2+, demonstrate that Q115A has a similar affinity towards -GAATTC- as to -GAAUTC-, while wild-type EcoRI binds to -GAATTC- with a tenfold preference over -GAAUTC-. On the basis of these thermodynamic and kinetic results it can be concluded that the hydrophobic contact between the gamma-methylene group of Gln115 and the methyl group of the inner thymidine contributes about 3 kJ/mol (0.7 kcal/mol) to the energy of interaction, both in the ground and the transition state. Q115E is catalytically inactive under normal buffer conditions, but becomes active at low ionic strength or in the presence of Mn2+. Different from Q115A, Q115E is inactive at alkaline pH and its DNA binding affinity is highest at acidic pH.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8421303 TI - Crystallization and preliminary X-ray crystallographic analysis of alpha-amylase from Bacillus subtilis. AB - Large crystals of alpha-amylase from Bacillus subtilis have been obtained at room temperature using polyethylene glycol 6000 as precipitant. They grow to typical dimensions of 0.25 mm x 0.3 mm x 2.0 mm in five days. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit cell dimensions of a = 85.46 A, b = 166.5 A and c = 332.7 A. The asymmetric unit seems to contain eight molecules of alpha-amylase, with crystal volume per protein mass (Vm) of 2.69 A3/Da and solvent content of 54.3% by volume. Despite a very long c-axis, the crystals diffracted to about 2.2 A Bragg spacing using the rotating anode X-rays and were resistant to damage by X-rays. Thus they are suitable for structure determination by X-ray methods at high resolution. X-ray diffraction data have been collected to 3.4 A Bragg spacing from a native crystal. PMID- 8421304 TI - Purification, crystallization and space group determination of DNA repair enzyme exonuclease III from E. coli. AB - Escherichia coli exonuclease III possesses multiple catalytic activities: (1) a nucleotidyl hydrolase activity cutting 5' to apurinic/apyrimidinic sites and urea residues in DNA; (2) a 3' to 5' exonuclease activity specific for double-stranded DNA; (3) a RNase H activity preferentially degrading the RNA strand of a DNA.RNA hybrid and (4) an activity that can remove a number of 3' termini from duplex DNA including 3' phosphates, 3' phosphoglycolate residues, 3' phosphoglycolaldehyde residues and 3' trans-4-hydroxy-2-pentenal-5-phosphate residues. These multiple activities make exonuclease III a major enzyme in the base excision repair pathway for DNA damage. We have purified exonuclease III and grown crystals by the vapor diffusion method using polyethylene glycol 4000 as the precipitant. Buffers were found to have profound effects on crystallization with high concentrations of imidazole/malate buffer (0.4 M to 1.0 M) yielding larger crystals with less twinning. The crystals belong to the space group P3(1)21 or its enantiomorph P3(2)21 with unit cell dimensions of a = b = 107.8 A, c = 42.2 A, alpha = beta = 90 degrees, gamma = 120 degrees, have one 31 kDa monomer per asymmetric unit and diffract to 1.6 A. These crystals are stable to X-rays and suitable for high resolution structure determination. PMID- 8421305 TI - Crystallization and preliminary crystallographic characterization of the copper containing amine oxidase from pea seedlings. AB - The copper-containing amine oxidase from pea seedlings has been crystallized using lithium sulfate as precipitant at pH 5.2. The unit cell is orthorhombic, space group P2(1)2(1)2(1), with dimensions a = 89.3 A, b = 113.4 A, c = 199.0 A. The mass of the asymmetric unit is 131(+/- 13) kDa, consistent with independent evidence that the molecule has two approximately 66 kDa subunits. The crystals diffract to 2.5 A in a synchrotron X-ray beam. PMID- 8421306 TI - Crystallization and preliminary X-ray analysis of the catalytic domain of xylanase a from Pseudomonas fluorescens subspecies cellulosa. AB - The catalytic domain of the xylan-degrading enzyme xylanase A, from Pseudomonas fluorescens subspecies cellulosa, has been expressed in Escherichia coli and crystallized. The crystals are well ordered and diffract to 1.8 A using X-rays generated at the Photon Factory in Japan. The crystals are orthorhombic, space group P2(1)2(1)2(1) with a = 95.7 A, b = 97.1 A and c = 149.8 A (all +/- 0.2 A). The similarity of the a and b cell edges, the intensity of the reflections along c* and the self rotation function results suggest a pseudo-tetragonal arrangement of molecules in the unit cell. There are probably four molecules in the asymmetric unit. PMID- 8421307 TI - Crystallization and preliminary X-ray analysis of human antithrombin III. AB - Human antithrombin III has been crystallized from 18 to 21% (w/v) polyethylene glycol 4000 at pH 7.15. The spacegroup is P2(1) with cell parameters a = 89.8 A, b = 100.8 A, c = 70.0 A and beta = 106 degrees. The diffraction limit is 3.2 A. The asymmetric unit contains two protein molecules. Analysis of dissolved crystals for biological activity and by gel electrophoresis suggests that one protein molecule in the asymmetric unit is intact, while the other is cleaved. PMID- 8421308 TI - Crystallization of the C-terminal domain of rabbit serum hemopexin. AB - The C-terminal domain of rabbit serum hemopexin, comprising residues 215 to 435, has been crystallized following removal of the attached carbohydrate using the endoglycosidase Endo F. The crystals, grown by vapour diffusion from solutions containing polyethylene glycol 1500, are orthorhombic, with cell dimensions a = 41.0 A, b = 64.2 A, c = 85.2 A, space group P2(1)2(1)2(1), and one molecule in the asymmetric unit. The crystals diffract to 2.4 A resolution and are suitable for X-ray structure analysis. PMID- 8421309 TI - Preliminary crystallographic analysis of sweet potato beta amylase. AB - Beta amylase from sweet potato has been crystallized in a form suitable for high resolution X-ray diffraction analysis from a mixture of polyethylene glycol 400 and ammonium sulfate at room temperature. The crystals are rectangular prisms and occasionally reach a size of 1 mm on an edge. The space group of the crystals is I422 with cell dimensions of a = b = 196.4 A and c = 72.2 A. There are four tetramers of about M(r) = 210,000 in the unit cell centered at 222 symmetry points. The asymmetric unit is a single beta amylase subunit of about M(r) = 52,000. The tetrameric molecule must have exact 222 molecular symmetry relating its subunits. PMID- 8421310 TI - Crystallization and preliminary X-ray crystallographic analysis of probable amylase/protease inhibitor-B from rice seeds. AB - Large crystals of probable amylase/protease inhibitor-B have been grown at room temperature from ammonium sulfate solution. The crystals grow within five days to dimensions of 0.6 mm x 0.6 mm x 0.6 mm. They diffract to at least 1.7 A upon exposure to synchrotron X-rays. The crystals belong to the space group P4(1)2(1)2 (or P4(3)2(1)2) with unit cell dimensions of a = 38.02 A and c = 98.98 A. The presence of one molecule per asymmetric unit gives the unit cell volume per protein mass (Vm) of 1.99 A3/Da and the solvent fraction of 38.2% by volume. X ray data have been collected to 2.0 A Bragg spacing from native crystals. PMID- 8421311 TI - Crystallization and preliminary X-ray diffraction analysis of ScrY, a specific bacterial outer membrane porin. AB - The sucrose-specific outer membrane porin ScrY of Salmonella typhimurium was isolated from Escherichia coli K-12 strain KS 26 containing the plasmid pPSO112. The protein was purified to homogeneity by differential extraction of the cell envelope in the presence of the detergents sodium dodecyl sulfate and lauryl (dimethyl)-amine oxide (LDAO). The porin had apparent molecular weights of 58 kDa and 120 kDa for the monomer and for the trimer, respectively, on SDS/PAGE. The purified trimers were crystallized using poly(ethylene glycol) 2000 and the detergents octylglucoside (OG) and hexyl-(dimethyl)-amine oxide (C6DAO). X-ray diffraction of the crystals showed reflections to 2.3 A. The space group of the crystals was R3 and the lattice constants of the hexagonal axes were a = b = 112.85 A and c = 149.9 A. The crystal volume per unit of protein molecular weight was 3.47 A3/Da. PMID- 8421312 TI - Critical role of the acceptor stem of tRNAs(Met) in their aminoacylation by Escherichia coli methionyl-tRNA synthetase. AB - To be aminoacylated by Escherichia coli methionyl-tRNA synthetase, a tRNA requires the presence of the methionine anticodon (CAU sequence). However, the importance in this reaction of the other nucleotides of tRNAs(Met) has still to be described. In this work, through the study of more than 35 variants of tRNAs(Met), it is shown, firstly, that the parameters of the aminoacylation reaction remain independent of the mutations affecting either the sequences or the sizes of the D-loop, D-stem and variable loop. This conclusion is illustrated by the construction and study of a tRNAf(MetCAU) with the D-stem, D-loop and very long variable loop of a class II tRNA. The resulting chimaeric tRNA is methionylated as efficiently as tRNAf(MetCAU) or tRNAm(MetCAU). Secondly, mutations affecting base 73 and base pairs 2.71 and 3.70 in the acceptor stem of tRNAf(MetCAU), as well as bases 32, 33 and 37, adjacent to the anticodon, cause a strong reduction of the rate of the aminoacylation reaction. Thirdly, it is shown that, provided it is given the acceptor stem of tRNAm(MetCAU) or tRNAf(MetCAU), a tRNA having the anticodon loop of tRNA(Met) can be converted into a substrate for methionyl-tRNA synthetase as efficient as tRNAf(MetCAU) or tRNAm(MetCAU). Finally, it is proposed that, beyond the binding of the anticodon loop to the synthetase, the sequence of the acceptor stem may strongly influence the rate of the catalytic step of the aminoacylation reaction by properly orientating the 3' end of the tRNA towards the catalytic centre. PMID- 8421313 TI - Primary structure of three distinct isoabrins determined by cDNA sequencing. Conservation and significance. AB - A family of toxic proteins, the isoabrins, which possess N-glycosylase activity toward eukaryotic 28 S r-RNA, may have potential use in cancer chemotherapy. By polymerase chain reaction techniques, cDNA clones of three isoabrins, carrying A and B-chain sequences, were isolated and their nucleotide sequences were determined. The isoabrins consist of an A-chain comprised of 250 of 251 amino acids, followed by a 10 amino acid linker and a B-chain of 267 amino acids. There is substantial conservation in the B-chain of the three isoabrins, with less than six amino acid substitutions, whereas as many as 46 amino acid substitutions occur in the A-chains. Based on the relationships between the biological activities and the putative amino acid sequences of the isoabrins, three isoabrins, abrin-a, -b and -d, could be identified and the potential epitope of immunological response of these isoabrins could be assigned. PMID- 8421314 TI - Isolation and sequencing of Escherichia coli gene proP reveals unusual structural features of the osmoregulatory proline/betaine transporter, ProP. AB - Transporters encoded in genetic loci putP, proP and proU mediate proline and/or betaine accumulation by Escherichia coli K-12. The ProP and ProU systems are osmoregulatory. Activation of ProP in response to hyperosmotic stress has been demonstrated both in vivo and in vitro. It therefore serves as a model experimental system for the analysis of osmosensory and osmoregulatory mechanisms. We developed methodologies which will facilitate the identification of proline transporter genes by functional complementation of putP proP proU bacteria. E. coli gene proP was isolated and located within a chromosomal DNA fragment. Deletion, complementation and sequence analysis revealed putative promoter and transcription termination signals flanking a 1500 base-pair open reading frame. The predicted 55 kDa ProP protein was hydrophobic. In vitro expression of proP yielded a protein whose apparent molecular mass was determined to be 42 kDa by polyacrylamide gel electrophoresis under denaturing conditions. Database searches and cluster analysis defined relationships among the ProP sequence and those of integral membrane proteins that comprise a transporter superfamily. Members of the superfamily catalyze facilitated diffusion or ion linked transport of organic solutes in prokaryotes and eukaryotes. Multiple alignment revealed particularly close correspondence among the ProP protein, citrate transporters from E. coli and Klebsiella pneumoniae and an alpha ketoglutarate transporter from E. coli. The predicted ProP sequence differed from those closely similar sequences in possessing an extended central hydrophilic loop and a carboxyl terminal extension. Unlike other protein sequences within the transporter superfamily, the carboxyl terminal extension of ProP was strongly predicted to participate in formation of an alpha-helical coiled coil. These data suggest that the ProP protein catalyzes solute-ion cotransport. Its unusual structural features may be related to osmoregulation of its activity. PMID- 8421315 TI - Effects of a single base-pair deletion in the bacteriophage lambda PRM promoter. Repression of PRM by repressor bound at OR2 and by RNA polymerase bound at PR. AB - We have deleted a single base-pair in the -35 region of the bacteriophage lambda PRM promoter. The deletion (PRM delta 34) creates a better match of PRM to consensus, thereby substantially increasing the activity of the promoter in vitro and in vivo. Since the mutation also increases the overlap between OR2 and the 35 region of PRM, binding of repressor to OR2 no longer activates, but in fact represses PRM. Finally, the mutation decreases the distance between the PRM and PR transcription start sites from 82 to 81 base-pairs. As a consequence, the interaction of RNA polymerase with either promoter in vitro strongly inhibits open complex formation at the other. Kinetic analyses and DNase I protection assays lead to the surprising result that mutual inhibition is not due to steric occlusion. Both promoters can be occupied by RNA polymerase at the same time. Determination of KB and kf revealed that inhibition of PRM delta 34 by PR was manifest in a 100-fold decrease in the value of kf, but at the same time KB was increased tenfold. These data raise the possibility that RNA polymerase molecules bound at the two promoters contact and mutually stabilize each other and that this interaction subsequently inhibits a substep in the isomerization of closed to open complexes. In footprinting assays, each promoter is characterized by sites of enhanced cleavage when that promoter is occupied alone. These enhancements are substantially diminished when both promoters are occupied, suggesting that complexes of each promoter with RNA polymerase alter the structure of complexes formed at the other promoter. Assays of the effects of the delta 34 mutation in vivo indicate that interference between PRM and PR does not limit the rate of open complex formation at PRM in the cell. Apparently, transcription initiation clears the promoter rapidly enough that neither promoter is occupied a significant fraction of the time. PMID- 8421316 TI - Domain organization of the subunit of the Salmonella typhimurium flagellar hook. AB - The deduced amino acid sequences of the family of axial proteins of the bacterial flagellum possess N and C-terminal heptad repeats of hydrophobic amino acid residues, which suggests that these proteins all fold to form bundles of alpha helices (e.g. coiled coils). There is evidence that flagellin, which is one of the axial proteins, has an axially oriented bundle of alpha-helices that gives rise to the inner, rod-shaped domains seen in electron density maps. We present evidence that a second member of the family, the hook subunit, also has such an axially oriented, rod-shaped domain. In three-dimensional reconstructions from electron micrographs of the helical hook of Salmonella typhimurium, the rod shaped domain has a diameter of 18 A, which is that expected for a coiled coil. The corresponding domain in the flagellin subunit of the filament, however, is larger, having a diameter of 24 A suggesting a bundle of three or more alpha helices. In addition to the rod-shaped domain, the hook has two other domains. At a radius of 55 A is the middle spheroidal domain about 25 A in diameter and at a radius of 75 A is the outer ellipsoidal domain about 20 A by 30 A by 40 A. The flagellin subunit also has a middle and an outer domain although they appear different from those of the hook. This is no doubt a result of the lack of any sequence similarity of the hook and flagellin subunits, apart from the N and C terminal heptad repeats. Along the hook axis, there is a 25 A wide channel, which presumably serves in the export of hook and flagellin subunits in the assembly of the filament. There is a comparably sized channel in the filaments as deduced from electron micrographs. Thus, electron microscopy consistently finds a small channel, whereas in X-ray diffraction studies of the filament, the channel size appeared to be about 60 A. At a diameter of 60 A, the channel could pass the flagellin or hook subunit in its completely folded state, but if the channel is only 25 A in diameter, the subunit would have to be at least partially unfolded in order to pass through the channel. PMID- 8421317 TI - General antimutators are improbable. AB - Antimutator mutations reduce spontaneous mutation rates, at least at some sites and along some pathways. Antimutators have been found in several microbial systems since their initial discovery in bacteriophage T4, where they occur mainly among mutations of gene 43 (which encodes the viral DNA polymerase). The phage T4 antimutators are highly specific, often strongly reducing mutations rates but only along specific pathways, usually A.T-->G.C. They may fail to affect other pathways, such as G.C-->A.T, and may even accelerate mutation at yet other pathways, such as transversions (R.Y-->Y.R). Both enzymatic and evolutionary considerations suggest that it should be difficult to isolate strong, general antimutator mutations, that is, mutations that substantially lower the total spontaneous mutation rate over the entire genome without producing strongly deleterious side effects. This notion has been tested by measuring mutation rates over a target comprising several kilobases in a set of phage T4 antimutators. In each case, this rate was indistinguishable from or greater than the wild-type rate. A survey of reports describing antimutators in other microbes reveals that none are yet demonstrated to be general antimutators. PMID- 8421318 TI - Cloning and overexpression of the triosephosphate isomerase genes from psychrophilic and thermophilic bacteria. Structural comparison of the predicted protein sequences. AB - We focused on the temperature adaptation of triosephosphate isomerase (TIM; E.C. 5.3.1.1.) by comparing the structure of TIMs isolated from bacterial organisms living in either cold or hot environments. The TIM gene from psychrophilic bacteria Moraxella sp. TA137 was cloned and its nucleotide sequence determined. Its deduced amino acid sequence revealed 34% identity with the thermophilic bacteria Bacillus stearothermophilus TIM. Expression vectors were constructed and recombinant Moraxella TA137 and Bacillus stearothermophilus TIMs were overproduced and purified to homogeneity. Recombinant TIM inactivation constants (Ki), measured at various temperatures, compared to those of the mesophilic Escherichia coli recombinant TIM clearly show that Moraxella TA137 and B. stearothermophilus TIMs have respectively psychrophilic and thermophilic characteristics. To try to elucidate the structure-thermolability and structure thermostability relationship, factors affecting the overall stability of these two TIMs were examined, based on the alignment with the mesophilic chicken TIM, the three-dimensional structure of which is already known. From this comparison, it appears that the adaptability of TIM to high temperature is favored by better stabilizing residues for the helix dipole as well as better helix-forming residues whereas the adaptability of TIM to low temperature seems to reside in the nature of helix-capping residues. PMID- 8421319 TI - Conformational and nucleic acid binding studies on the synthetic nucleocapsid protein of HIV-1. AB - A 55 residue peptide corresponding to the nucleocapsid protein of HIV-1 (NCp7) containing two zinc binding domains as well as three truncated peptides were synthesized by Fmoc-based solid phase synthesis using the fragment condensation approach. Circular dichroism (CD) data support a conformational model in trifluoroethanol/buffer solution consisting of two helical segments at the chain ends with two Zn-modules in the center of the molecule. CD titration experiments show that the synthetic protein binds two equivalents of Zn2+ stoichiometrically, and the Zn2+ induced conformational changes are completely reversible by addition of EDTA. NCp7 and its S-acetamidomethylated analog (NCp7-Acm), devoid of the zinc co-ordination centers, exhibit preferential binding to RNA with a Kd = approximately 10(-9) M irrespective of the cysteine modification as determined by filter binding assays. The binding affinity of the NCp7 protein to single stranded DNA is lower than to RNA. Binding to double-stranded DNA is lower than to ssDNA. The NCp7-Acm protein exhibits reduced single-stranded DNA binding affinity compared to the unmodified protein. Nucleic acid binding analyses with the fragments of NCp7 protein suggest that two basic amino acid stretches are involved in RNA binding of the NCp7. PMID- 8421320 TI - Tissue distribution, excretion, and urinary metabolites of dichloroacetic acid in the male Fischer 344 rat. AB - The disposition of dichloroacetic acid (DCA) was investigated in Fischer 344 rats over the 48 h after oral gavage of 282 mg/kg of 1- or 2-[14C]-DCA (1-DCA or 2 DCA) and 28.2 mg/kg of 2-DCA. DCA was absorbed quickly, and the major route of disposition was through exhalation of carbon dioxide and elimination in the urine. The dispositions of 1- and 2-DCA at 282 mg/kg were similar. With 2-DCA, the disposition differed with dose in that the percentage of the dose expired as carbon dioxide decreased from 34.4% (28.2 mg/kg) to 25.0% (282 mg/kg), while the percentage of the radioactivity excreted in the urine increased from 12.7 to 35.2%. This percentage increase in the urinary excretion was mostly attributable to the presence of unmetabolized DCA, which comprised more than 20% at the higher dose and less than 1% at the lower dose. The major urinary metabolites were glycolic acid, glyoxylic acid, and oxalic acid. DCA and its metabolites accumulated in the tissues and were eliminated slowly. After 48 h, 36.4%, 26.2%, and 20.8% of the dose was retained in the tissues of rats administered 28.2 and 282 mg/kg of 2-DCA and 282 mg/kg of 1-DCA, respectively. Of the organs examined, the liver (4.9-7.9% of dose) and muscle (4.5-9.9%) contained the most radioactivity, followed by skin (3.3-4.5%), blood (1.4-2.6%), and intestines (1.0 1.7%). One metabolite, glyoxylic acid, which is mutagenic, might be responsible for or contribute to the carcinogenicity of DCA. PMID- 8421321 TI - Metabolic interaction of ethanol and cyclohexanone in rabbits. AB - The interaction of ethanol (EtOH) and cyclohexanone (CHN) metabolism was studied to demonstrate the influence of alcohol beverage in cyclohexanone poisoning. Rabbits were administered CHN and EtOH separately or together, and the plasma concentration of CHN and cyclohexanol (CHL), a metabolite of CHN, and blood concentrations of EtOH were measured at various times. When CHN alone was administered orally, the time to maximum plasma concentration (Tmax) of CHN was as short as 15 min, but that of CHL was 120 min. The short Tmax of CHN was considered to be due to the first-pass effect. The plasma concentration of CHL was much greater than that of CHN. On the other hand, when the same amount of molar CHL was administered in rabbits for sake of comparison, only small amounts of CHN were detected in plasma. The interconversion between CHN and CHL at the time of equilibration tended to shift predominantly toward the formation of CHL, whose plasma concentration ratio was about 1:6 to 1:8. The coadministration of EtOH decreased the plasma concentration of CHN and increased that of CHL more than the administration of CHN alone. The blood concentration of EtOH was also decreased with the coadministration of CHN. The coadministration of EtOH with CHN demonstrated an acceleration in the metabolism of both CHN and EtOH, which may be due to an increase in the concentrations of the alcohol dehydrogenase-NADH and NAD complexes. PMID- 8421322 TI - Alterations in cytochrome P-450 levels in adult rats following neonatal exposure to xenobiotics. AB - Neonatal exposure to certain xenobiotics has been shown to alter hepatic metabolism in adult rats in a manner that indicates long-term changes in enzyme regulation. Previously, we have observed changes in adult testosterone metabolism and in cytochrome P-450 (P-450) mRNA levels in animals neonatally exposed to phenobarbital (PB) or diethylstilbestrol (DES). In order to test for other enzyme alterations, we used Western blot procedures for specific P-450s to analyze hepatic microsomes from adult rats (24 wk old) that had been exposed neonatally to DES, PB, 7,12-dimethylbenz[a]anthracene (DMBA), or pregnenolone 16 alpha carbonitrile (PCN). The most striking effects were observed in the DES-treated males: P-4502C6 and an immunologically similar protein were increased 60 and 90%, respectively, relative to control values, but P-4503A2 was decreased by 44%. No changes were observed in the DES-treated males in levels of P-4502E1, P-4502B, or the male-specific P-4502C13. Adult males neonatally treated with PB had 150% increase in levels of anti-P4502B-reactive protein without significant changes in the other enzymes. The DES- and DMBA-treated females had increased levels of the female-specific P-4502C12 of 38 and 48%, respectively, but no other observed alterations. The results confirm that neonatal exposure to DES or PB can cause alterations in adult hepatic cytochrome P-450 levels but show that these chemicals act on different enzymes. Neonatal DMBA resulted in changes in adult females similar to those produced by the synthetic estrogen DES, but did so at about two-thirds lower dose. PMID- 8421323 TI - Subchronic oral toxicity study of captafol in B6C3F1 mice. AB - The effects of subchronic administration of captafol were studied in B6C3F1 mice given dose levels of 0, 0.3, 0.625, and 1.25% in the diet for 12 wk. There was a dose-related decrease in body weight gain during the 12-wk experiment and a loss of body weight in the 1.25% group of both sexes. Whiles the mice given captafol consumed less diet than the control mice, this was not directly dose-related. The relative weights of liver demonstrated a tendency for dose-dependent increase. Light-microscopic examination revealed cytoplasmic vacuolar degeneration, depending in severity on the dosage, in the livers of both sexes given captafol. In conclusion, the findings obtained from the present subchronic toxicity study indicated the liver to be a primary target organ. PMID- 8421324 TI - d-limonene exposure to humans by inhalation: uptake, distribution, elimination, and effects on the pulmonary function. AB - The toxicokinetics of d-limonene were studied in human volunteers exposed by inhalation (2 h, work load 50 W) in an exposure chamber on three different occasions. The exposure concentrations were approximately 10, 225, and 450 mg/m3 d-limonene. The relative pulmonary uptake was high, approximately 70% of the amount supplied. The blood clearance of d-limonene observed in this study, 1.1 l kg-1 h-1, indicates that d-limonene is metabolized readily. About 1% of the total uptake was eliminated unchanged in the expired air after the end of exposure, while approximately 0.003% was eliminated in the urine. A long half-time in blood was observed in the slow elimination phase, which indicates accumulation in adipose tissues. A decrease in vital capacity was observed after exposure to d limonene at a high exposure level. The subjects did not experience any irritative symptoms or symptoms related to the central nervous system (CNS). PMID- 8421325 TI - Acute and subacute toxicity tests of onion coat, natural colorant extracted from onion (Allium cepa L.), in (C57BL/6 x C3H)F1 mice. AB - The toxicity test of onion coat colorant (OC), a food colorant extracted from onion (Allium cepa L.), was undertaken using (C57BL/6 x C3H)F1 mice of both sexes for the safety assessment of this product. The acute toxicity test was performed by administration of OC suspended in corn oil by gavage at doses of 2500, 5000, 7500, and 10,000 mg/kg body weight to groups of 5 or 6 males and 6 or 7 females, maintained for 14 d. Six of 12 females dosed at 10,000 mg/kg body weight and 3 of 11 females dosed at 7500 mg/kg body weight were dead before the end of the study, indicating that the tolerated dose of OC was between 7500 and 5000 mg/kg body weight. The subacute toxicity test of OC was examined using 123 mice of both sexes (62 males and 61 females) by feeding a diet mixed with OC at concentrations of 5, 2.5, 1.25, 0.6, and 0.3% for 90 d. All mice tolerated these doses of OC well. The body weight gains of male and female mice were not affected by the treatment. Histopathological examinations showed that hyperplastic changes in the esophagus, forestomach, pancreas, cervix, and endometrium of mice were found in treated and control mice. However, their incidences were not related to the dose of OC. Moreover, only a spontaneous ovarian teratoma was found in an OC-treated mouse. These results suggest that OC has no acute and subacute toxic effects in mice. PMID- 8421326 TI - Axillofemoral bypass with externally supported, knitted Dacron grafts: a follow up through twelve years. AB - PURPOSE: The purpose of this study was to review our experience with externally supported, knitted Dacron grafts used for axillofemoral bypass. METHODS: Retrospective analysis was performed on records of 79 consecutive axillofemoral bypass graft operations performed on 77 patients from January 1978 to April 1990. RESULTS: The mortality rate within 30 days of operation was 5% (four of 79); 36 patients died in the follow-up period; none died of graft causes. During this 12 year period (mean follow-up 42 months) three patients were unavailable for follow up. The primary patency rate was 78% at 5 years and 73% at 7 years, with no change thereafter. Neither the graft configuration (i.e., axillounifemoral [n = 50] vs axillobifemoral [n = 29]) nor patency of the superficial femoral artery had an impact on the primary patency rate. Patients who underwent surgery for disabling claudication (n = 30 grafts) had a primary patency rate of 80% at 6 years compared with 65% at 6 years for those who required surgery for limb salvage (n = 49 grafts); the difference was not significant (p = 0.37). Actuarial survival of patients with axillofemoral grafts was 23% at 10 years compared with 72% in a concurrent population of patients with aortofemoral bypass (p < 0.001). CONCLUSION: These findings indicate that axillofemoral bypass grafts may be appropriate for high-risk patients with severe aortoiliac disease who require revascularization for either limb salvage or incapacitating claudication. PMID- 8421327 TI - Refining the indications for arteriography in penetrating extremity trauma: a prospective analysis. AB - PURPOSE: Five hundred fourteen consecutive patients with an isolated upper or lower extremity penetrating injury were entered into a prospective study designed to refine the indications for diagnostic arteriography. METHODS: Twenty-two (4%) patients with limb-threatening ischemia who required immediate operation and 23 (4%) who refused arteriography were excluded from subsequent analyses. The remaining 469 patients were classified as being at high, intermediate, or low risk for an arterial injury. RESULTS: Two hundred thirteen patients who were at low risk were observed for 24 hours, discharged, and monitored as outpatients. No delayed complications of an arterial injury developed in any patient in this group. The intermediate-risk group of 151 patients and the high-risk group of 105 patients underwent arteriography. Seventy-seven injuries were identified; 24 were major (limb-threatening) and 53 were minor. Fourteen major injuries required operative repair or transcatheter embolization; the remaining 10 nonocclusive major injuries were observed without sequelae. CONCLUSIONS: By step-down logistic regression only pulse deficit (p < 0.01) and an ankle/brachial or wrist/brachial index less than 1.00 in the injured extremity (p < 0.03) were found to be significant predictors of an arterial injury. The presence of either of these two clinical variables successfully predicted all major arterial injuries. This prospective study supports the proposition that arteriography that is limited only to those patients who have either a pulse deficit or minimum ankle/brachial or wrist/brachial index less than 1.00 successfully detects all significant arterial injuries. PMID- 8421328 TI - A prospective study of the clinical outcome of femoral pseudoaneurysms and arteriovenous fistulas induced by arterial puncture. AB - PURPOSE: Although spontaneous thrombosis of femoral false aneurysms (FAs) and arteriovenous fistulas (AVFs) has been reported, the frequency of this occurrence is unknown. This prospective study was designed to establish the natural history of FA and AVF and to evaluate factors that might predict eventual thrombosis of these lesions. METHODS: Twenty-two patients with either femoral FAs (n = 16) or AVFs (n = 6) induced by percutaneous arterial punctures were evaluated prospectively. After an initial duplex scan, all patients were monitored with serial scans, either in hospital or weekly as outpatients, depending on the stability of the process. Operative repair was performed for the following indications: (1) a greater than 100% increase in size of a FA by duplex scan, (2) the development of symptoms, or (3) continued patency of the lesion after 2 months of observation. RESULTS: Nine of 16 FAs and four of six AVFs closed spontaneously; FAs greater than 6 cm3 (1.8 cm in diameter) required repair more often (p = 0.065). However, size was not an absolute predictor of the need for repair because two small aneurysms (1.6 and 0.7 cm3) remained patent, although both patients were discharged safely from the hospital, and two large aneurysms (13.2 and 10.7 cm3) thrombosed spontaneously. Three of seven patients whose aneurysms required repair received anticoagulation continuously from the time of catheterization until repair became necessary. None of the patients whose FAs closed spontaneously were receiving anticoagulants at the time of thrombosis (p = 0.02). Neither length of the FA neck, velocity in the FA cavity, size of original arterial puncture, nor velocity in the AVF correlated with thrombosis. CONCLUSIONS: We conclude that (1) all FAs do not thrombose spontaneously and at least one third require surgical repair, (2) patients receiving continuous anticoagulation should undergo aneurysm repair, (3) discharge of patients with FAs less than 6 cm3 is safe (the majority of these FAs will eventually thrombose spontaneously), and (4) many AVFs close spontaneously and repair is not required unless symptoms or signs of progressive enlargement develop. PMID- 8421329 TI - Innominate artery trauma: a thirty-year experience. AB - PURPOSE: Injury to the innominate artery may represent a zone I cervical, thoracic outlet, or intrathoracic vascular injury and poses diagnostic, exposure, and management problems for the general, vascular, and thoracic surgeon. This complex injury often becomes a new learning experience with each infrequent encounter. METHODS: Between 1960 and 1992 43 patients with innominate artery injuries were treated. Penetrating injuries were from gunshot wounds in 25, stab wounds in 7, and shotgun wounds in 2 patients. Blunt injuries accounted for seven of the patients. In 28 patients there were multiple injuries, including multiple vascular injuries. Median sternotomy was used in all planned operations in the later part of the study, and bilateral transsternal anterolateral thoracotomy was used in patients undergoing urgent or emergency center thoracotomy. RESULTS: Blunt injury resulted in tears near the aortic arch with intimal disruption. Bypass grafting without hypothermia, shunts, or systemic heparinization is now used in all patients. Thirty-two patients survived to leave the hospital with no new complications related to the procedure. Postoperative neurologic complications were associated with preexisting neurologic deficits. CONCLUSIONS: Patients with innominate artery injury and stable vital signs can usually be transported without difficulty and treated without complex perioperative adjuncts. These patients can undergo revascularization with simple vascular surgical techniques and should expect an uncomplicated postoperative course unless there has been associated central nervous system injury or related injuries leading to systemic infection. Synthetic conduits have been used with success and have not required systemic heparinization or complex temporary shunting. PMID- 8421330 TI - Carotid endarterectomy with reconstruction techniques tailored to operative findings. AB - PURPOSE: Reconstruction techniques tailored to operative findings were applied to 466 consecutive carotid endarterectomies (CEA) performed on 408 patients over 5 years. The choice of reconstructive technique was based on the extent of the arteriotomy incision required to obtain a complete internal CEA endpoint, the ability to obtain a complete endpoint, and the quality or redundancy of the endarterectomized internal carotid artery (ICA) segment. The hypothesis was that a complete internal CEA endpoint and a tapered, smooth, nonkinked reconstruction minimize complications. METHODS: Complete distal endpoint feathering was obtained in 437 (94%) CEA. Patch reconstruction was performed in the 429 (92%) CEA in which the arteriotomy extended distal to the ICA bulb. Before patching, 16 (3.4%) redundant endarterectomized ICA segments were shortened by transverse-eversion suture plication to prevent kinking. A saphenous vein interposition graft was used in nine (1.9%) CEA. The other 28 (6.0%) CEA had an arteriotomy that did not extend distal to the bulb and were primarily closed. RESULTS: Two (0.4%) patients died of myocardial infarction in the hospital; one of these patients also had a stroke. Three (0.6%) patients had nonfatal strokes in the hospital. Five patients had hyperperfusion syndrome after CEA, one of which occurred without complications in the hospital. The other four occurred after discharge from the hospital. Three of these patients had a stroke, and two with strokes died. The combined 30-day mortality and nonfatal stroke rate was 2.1% for CEA and 2.4% for patients. There were no patch ruptures, false aneurysms, or ICA occlusions. Three patients had a > 50% diameter carotid artery stenosis 6 months after CEA. CONCLUSIONS: These results support an aggressive attempt to obtain a complete or optimal ICA endpoint with reconstruction techniques based on operative findings. Recognition of patients at risk for and treatment of hyperperfusion syndrome after CEA remains a clinical challenge. PMID- 8421331 TI - Is the iliac artery a suitable inflow conduit for iliofemoral occlusive disease: an analysis of 514 aortoiliac reconstructions. AB - PURPOSE: The aorta is the conventional inflow source for reconstructions in patients with aortoiliofemoral occlusive disease. In patients with unilateral iliac or femoral disease, femoral-to-femoral bypasses have been used but with less favorable patency rates. The purpose of this study is to evaluate the performance of the unobstructed iliac artery as an inflow source for ipsilateral, contralateral, or bilateral reconstructions in iliofemoral occlusive disease. METHODS: Over the past 6 years 322 reconstructions have been performed with the iliac artery as the donor vessel. Patients were evaluated for proximal hemodynamically significant lesions by augmented pullout pressures during aortography. Patients who had balloon angioplasty were excluded. RESULTS: Results were compared with 192 patients who underwent conventional aortodistal bypass operation for occlusive disease during the same period. Both groups were similar in risk factors, age, sex, and indications for operation. For the iliac group the operative mortality rate was 1.6%, and the 30-day patency rate was 97%, similar to those in the aortic group (3.6% and 95%, respectively). Cumulative patency rates at 5 years by life-table analysis were 82% for iliac artery inflow and 77% for aortic inflow reconstructions. CONCLUSIONS: Our experience suggests that an unobstructed iliac artery is a reasonable inflow source for reconstructions in iliofemoral occlusive disease. The long-term patency rate is comparable to aortodistal bypasses and superior to other extraanatomic bypasses. PMID- 8421332 TI - Correlation of North American Symptomatic Carotid Endarterectomy Trial (NASCET) angiographic definition of 70% to 99% internal carotid artery stenosis with duplex scanning. AB - PURPOSE: The North American Symptomatic Carotid Endarterectomy Trial (NASCET) has thus far demonstrated conclusive benefit for carotid endarterectomy for patients with symptomatic 70% to 99% internal carotid artery (ICA) stenosis. In the NASCET, ICA stenosis was classified angiographically: % ICA stenosis = (1 - [narrowest ICA diameter/diameter normal distal cervical ICA]) x 100%. However, widely used duplex scan criteria for ICA stenosis correlate with different angiographic categories of high-grade stenosis (50% to 79%, > 80%) and were developed on the basis of estimated bulb diameter. We therefore blindly evaluated with separate observers carotid angiograms from 100 patients who also underwent carotid duplex scanning in our vascular laboratory. METHODS: "Angiographic stenosis" was calculated as in NASCET. Duplex scan measurements of ICA peak systolic velocity (PSV), ICA end-diastolic velocity, and the ratio of ICA PSV to common carotid artery (CCA) PSV were analyzed for sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy to identify a 70% to 99% ICA stenosis. RESULTS: Analysis of the data revealed that an ICA PSV/CCA PSV ratio of 4.0 provided the best combination of sensitivity (91%), specificity (87%), positive predictive value (76%), negative predictive value (96%), and overall accuracy (88%) for detection of a 70% to 99% stenosis. CONCLUSION: We conclude duplex scan determination of 70% to 99% stenosis as defined in the NASCET requires the adoption of duplex criteria modified from those in current use in most vascular laboratories. PMID- 8421333 TI - The natural history of asymptomatic carotid artery disease. AB - PURPOSE: The purpose of this article is to determine the natural history of carotid artery disease among asymptomatic patients with cervical bruits or other risk factors for stroke and to study the value of duplex ultrasonography in predicting future neurologic events. METHODS: Two hundred forty-two asymptomatic, unoperated patients, referred for evaluation of asymptomatic carotid artery disease, were followed prospectively with duplex ultrasonography. RESULTS: Fifteen ischemic strokes (6.2%) and 20 transient ischemic attacks (TIA) (8.3%) occurred in 34 patients during a mean follow-up of 27.4 months. Annual stroke, TIA, and combined event rates were 2.7%, 3.6%, and 6.2%, respectively. Although patients with 80% to 99% lesions had a 20.6% annual event rate, most events occurred contralateral to these lesions; the vessel-specific annual event rate for 80% to 99% disease was 5.1%. Only one of 15 strokes occurred ipsilateral to an 80% to 99% stenosis. Echolucent plaques were associated with TIA and stroke (5.7% annual vessel event rate vs 2.4% for echogenic plaques, p = 0.03). Disease progression was highly correlated with TIA and stroke (p < 0.0001), but it usually occurred in association with rather than before ischemic events, thus proving more useful in explaining pathogenesis than in predicting future events. There was no association between aspirin use and TIA, but patients taking aspirin had a threefold higher annual stroke rate (1.6% vs 4.8%, p = 0.027). CONCLUSIONS: This study, while confirming significant risk for asymptomatic patients with critical stenosis or echolucent plaque, demonstrates the importance of contralateral disease and the absence of orderly progression from minimal disease through high-grade stenosis to symptomatic cerebral ischemia. TIA and stroke commonly occur in association with abrupt, unpredictable, quantum changes in carotid artery disease. PMID- 8421334 TI - Nonoperative treatment of superficial femoral artery disease: long-term follow up. AB - PURPOSE: Between 1977 and 1991, 405 patients with atherosclerotic occlusive disease of the superficial femoral artery underwent clinical as well as noninvasive laboratory evaluation and were recommended for nonoperative treatment. METHODS: Limbs with uncorrected aortoiliac occlusive disease, aneurysmal degeneration, or previous femoropopliteal bypass were excluded, leaving 568 involved extremities. Complete follow-up, which forms the basis for this report, was available in 377 patients (93%) with 520 limbs (93%). Patients were monitored for a minimum period of 2 years (range, 24 to 164 months; median, 86 months). During the surveillance period 45 limbs (8.6%) in 42 patients (11.1%) required arterial intervention. This entailed operation in 39 cases and endovascular treatment in six cases. With use of life-table analysis, the risk for intervention was found to be 11% at 5 years and 14% at 10 years. A total of 14 limbs (2.7%) in 14 patients (3.7%) ultimately required major limb amputation, either after failed bypass (8 patients) or as a primary procedure (6 patients). RESULTS: Analysis of risk factors revealed that female sex (p = 0.04), chronic renal failure (p = 0.0001), diabetes mellitus (p = 0.0011), history of contralateral femoropopliteal bypass (p = 0.0005), level of disease (p = 0.003), and entry ankle/brachial index less than 0.50 (p = 0.004) were associated with an increased risk over time for intervention. Other factors, including age, current or prior smoking history, hypertension, and the presence of coronary artery disease or cerebrovascular disease failed to reach statistical significance. CONCLUSIONS: These data support the continued conservative approach to surgery for patients with superficial femoral artery occlusive disease without limb threatening symptoms. Patients with multilevel disease, lower ankle/brachial index, a history of contralateral femoropopliteal bypass, chronic kidney failure, or diabetes mellitus are at increased risk and should be monitored more closely. PMID- 8421335 TI - Risks and benefits of femoropopliteal percutaneous balloon angioplasty. AB - PURPOSE: The purpose of this study was to evaluate the efficacy of angioplasty in the treatment of femoropopliteal arterial disease. METHODS: From 1980 to 1991, 126 angioplasty procedures were performed in 131 limbs of 106 patients with 175 femoropopliteal lesions (26 common femoral, 118 superficial femoral, and 31 popliteal). Critical ischemia was present in 55 limbs (42%), and claudication was present in 76 (58%). Angioplasty was performed for a single lesion in 87 limbs (66%) and for multiple lesions in 44 (34%). In 13 limbs (10%) the most severe lesion was an occlusion; in 118 (90%) all lesions were stenoses. Distal runoff was good (2 or 3 vessels patent) in 72 limbs (55%) and poor (0 or 1 vessel patent) in 59 (45%). RESULTS: Death within 30 days occurred in 0.8%, nonfatal systemic morbidity in 7.1%, and local morbidity in 1.6% of procedures. Multivariate analysis revealed that indication and age were predictive of increased morbidity and mortality rates. Immediate success was achieved in 95% of limbs treated. Mean follow-up time was 2.0 years. The overall 5-year cumulative primary patency rate was 45% (+/- 5%). In a proportional hazards model indication and lesion type were predictive (p < 0.01) of long-term failure, with relative risks of 2.0 (1.2 to 3.3) and 2.7 (1.3 to 5.6), respectively. The 5-year primary patency rate after angioplasty for stenoses and claudication was 55% (+/- 7%), for stenoses and critical ischemia it was 29% (+/- 11%), and for occlusions it was 36% (+/- 14%). CONCLUSION: These results suggest that femoropopliteal angioplasty is a low-risk procedure with acceptable long-term results in patients with claudication and stenoses. PMID- 8421336 TI - The characteristics and anatomic distribution of lesions that cause reversed vein graft failure: a five-year prospective study. AB - PURPOSE: The cause of vein graft failure in the intermediate postoperative period (3 to 18 months) has not been well defined. To delineate the incidence, characteristics, and anatomic distribution of lesions that cause graft failure in this critical interval, 227 consecutive infrainguinal reversed vein grafts (IRVGs) constructed at a single institution from July 1986 to December 1991 were prospectively entered into a duplex scan surveillance protocol. METHODS: Duplex surveillance with arteriographic confirmation identified 29 patent, hemodynamically failing IRVGs during a mean follow-up of 22 months (range 1 to 64 months). An additional 18 grafts thrombosed before detection of any underlying abnormality; thrombolytic therapy and repeat operation uncovered the cause of occlusion in 12 of these grafts. The cause of graft failure (failing as well as failed) was therefore clear in 41 (87.2%) of 47 instances. RESULTS: The causes of failure were intrinsic graft stenosis (n = 28; 59.6%), inflow failure (n = 6; 12.8%), outflow failure (n = 4; 8.5%), muscle entrapment (n = 2; 4.3%), and hypercoagulable state (n = 2; 4.3%). The most common intrinsic graft lesion was focal intimal hyperplasia (18 lesions in 16 grafts) in the juxtaanastomotic position, occurring solely in the vein graft itself. It occurred with equal frequency immediately distal to the proximal anastomosis or proximal to the distal anastomosis. Only rarely (n = 5) did this involve the juxtaanastomotic artery. Focal midgraft valvular stenoses (n = 6) and diffuse myointimal hyperplasia (n = 4) were also detected. The peak incidence of graft failure was 4 to 12 months after operation (70% within 12 months, 80% within 18 months). CONCLUSIONS: We conclude that duplex surveillance of IRVGs is warranted by the 21% incidence of potentially remediable graft failure. A significant portion of these failures occur during the intermediate postoperative period (3 to 18 months), usually as a result of focal intrinsic vein graft lesions. With reversed vein conduits, these lesions arise predominantly in the vein graft itself, in the juxtaanastomotic position. PMID- 8421337 TI - Biologic characteristics of long-term autogenous vein grafts: a dynamic evolution. AB - PURPOSE: The venous conduit as an arterial substitute has dynamic biologic properties that affect its durability. This study evaluated the morphologic and physiologic characteristics of 72 lower extremity vein grafts functioning at 4.5 to 21.6 years (median 6.6 years). METHODS: The entire graft was imaged with use of color duplex ultrasonography and then classified as normal (class I), abnormal but not graft-threatening (class II), or abnormal and graft-threatening (class III) for the proximal, middle, and distal thirds. Thirty-one grafts (43%) were classified as normal, whereas 41 (57%) were classified as abnormal, with 58 class II and 15 class III segments. RESULTS: Three types of abnormalities were found by duplex imaging: nonstenotic wall plaques, discrete stenoses, and aneurysmal dilation. Aneurysms developed in five of seven grafts that had required thrombectomy in the distant past (mean of 40 months). There were 70 postoperative revisions in 38 limbs (53%). In 23 (60%) the conduit was revised, in 11 (29%) the revisions corrected progression of native artery atherosclerotic disease, and in 4 (11%) both types of revisions were required. Eleven grafts were revised in the first 30 days to correct technical errors. Eighteen limbs were revised between 1 and 24 months, with 12 (67%) of the revisions correcting stenotic lesions in the conduit or at one of the anastomoses. After 24 months 12 (67%) of 18 limbs were revised to correct progression of occlusive disease in the inflow or outflow vessels. At the time of this study 18 (67%) of the 27 conduits revised for intrinsic lesions were abnormal by color duplex imaging, and they harbored 12 (80%) of the 15-graft-threatening lesions. CONCLUSIONS: Autogenous vein remains the most durable arterial conduit, but vigilant surveillance is essential because the atherosclerotic environment continually produces lesions that may imperil the longevity of the graft. PMID- 8421338 TI - Surgical treatment of aneurysm of the persistent sciatic artery. AB - Persistent sciatic artery (PSA) is a rare congenital malformation that is complicated by aneurysm formation in more than 25% of the reported cases. Two cases of aneurysm of the PSA are presented. Twenty-six aneurysms of the PSA, including our two cases, have been reported in the English literature in the last three decades. Early surgical treatment is warranted to decrease the 25% amputation rate associated with thromboembolic complications. The posterior, transgluteal repair of this aneurysm affords excellent exposure and avoids a long bypass graft, multiple incisions, and staged procedures. Magnetic resonance imaging may be helpful in preoperative evaluation of the feasibility of proximal, extrapelvic vascular control. PMID- 8421339 TI - Transcutaneous ultrasonography can be used to guide and monitor balloon angioplasty. AB - PURPOSE: Percutaneous balloon angioplasty is an accepted technique for the treatment of short segmental stenoses of the iliac and superficial femoral arteries. Some surgeons have not embraced this technique because of lack of training, unfamiliarity with radiologic equipment, or poor-quality fluoroscopy equipment in the operating room. A new technique, ultrasound-directed balloon angioplasty, enables the vascular surgeon to guide the catheter and evaluate the progress of the procedure without the use of radiographic imaging. METHODS: The new catheter is integrated externally into a standard duplex scanner. A spherical brass bead positioned on the catheter shaft in the middle of the balloon is covered with a piezoelectric material. This omnidirectional receiver is connected through a wire to a catheter interface system, which allows the exact position of the balloon to be represented on the screen of a duplex unit. The catheter is advanced under visual control with use of the scanner. RESULTS: The system has been used to perform balloon angioplasty on 21 lesions (16 superficial femoral, 3 iliac, 2 popliteal arteries) in 17 patients. Procedures in 16 patients were performed percutaneously; in an additional patient bilateral angioplasties were performed during operation. The balloon was successfully positioned in each case with use of ultrasonography alone; each procedure was confirmed by use of angiography. Angioplasty was accomplished in 16 (76%) of 21 with the ultrasound guided catheter alone. Supplemental use of a high-pressure balloon or atherectomy device was necessary in five patients. CONCLUSIONS: This technique records physiologic and anatomic data in real time so that the progress of the angioplasty can be monitored. In three patients the inadequacy of the initial angioplasty was recognized by a persistent velocity increase but not by angiography. As a result corrective measures were taken. This new procedure allows vascular surgeons to use equipment with which they are familiar, reduces the risk of ionizing radiation and contrast nephropathy, and permits the monitoring of the angioplasty with hemodynamic parameters. PMID- 8421340 TI - Infrainguinal reconstruction for patients with chronic renal insufficiency. AB - PURPOSE: The efficacy of autogenous infrainguinal reconstruction for patients with varying degrees of chronic renal insufficiency was reviewed because it has not been well defined. METHODS: Pertinent data were retrieved from hospital records and the vascular registry for all patients with chronic renal insufficiency (serum creatine > 2 mg/dl) who required surgical intervention for ischemic lower limbs during the past 15 years at Brigham and Women's Hospital. RESULTS: During the past 15 years, 56 patients underwent 70 autogenous vein bypass procedures, and 31 underwent 42 primary major amputations. All patients had serum creatinine levels greater than 2 mg/dl. Limb salvage was the indication for intervention in 84% of reconstructed limbs, and 48% required infrapopliteal bypass. No significant difference was found in the operative mortality rate associated with primary amputation (17%) and reconstruction (11%), but overall 5 year survival for the group undergoing reconstruction (40%) was significantly superior to that observed in the group undergoing amputation (9%). The 5-year cumulative limb salvage rate for the group having reconstruction was 80%, with a primary graft patency of 74% and a secondary patency of 77%. Patients with diabetes had lower rates of both limb salvage (63%) and survival (21%). No patient on dialysis survived beyond the third-year interval, but the limb salvage rate after 2 years was 76%. CONCLUSION: Patients with chronic renal insufficiency achieved graft patency and limb salvage results comparable to those obtained in patients with normal kidney function, but they sustained higher perioperative morbidity and reduced survival. PMID- 8421341 TI - Impact of a color-flow duplex surveillance program on infrainguinal vein graft patency: a five-year experience. AB - PURPOSE AND METHODS: The contribution of color-flow duplex surveillance to improving vein graft patency was evaluated in two patient groups after 201 infrainguinal bypass procedures were performed during a 5-year period. Incidence of revision procedures and the primary and assisted primary patency rates were compared for 160 bypass grafts monitored during the first 2 years by use of color flow duplex scanning of the vein graft and adjacent arterial segments (color-flow surveillance group) versus 41 bypass grafts monitored by use of clinical assessment alone (clinical follow-up group). Only grafts that were patent after the first postoperative month are considered. RESULTS: The two groups were comparable with regard to most of the pertinent clinical factors. Stenotic lesions were identified in 58 bypass grafts, and severity was determined by use of intraarterial digital subtraction angiography. Eighteen bypass grafts with stenoses did not undergo a revision for reasons that were determined by the doctor, the hospital, or the patient. The occlusion rates of revised and nonrevised stenotic grafts were compared for lesions of different severity. None of the grafts for stenoses with 30% to 49% diameter reduction (DR) failed during follow-up. Occlusion occurred in 57% of the nonrevised and 9% of revised grafts (p = 0.047) for stenoses with 50% to 69% DR. Stenoses with 70% or greater DR were associated with graft failure in 100% of nonrevised bypasses and in 10% of revised grafts (p = 0.004). The assisted primary patency rate was higher in grafts that underwent color-flow surveillance compared with grafts with that underwent clinical follow-up (3-year patency rates of 91% and 72%, respectively; p = 0.004). The independent correlation of color-flow surveillance with higher patency rates was demonstrated in a proportional hazard analysis. The relative risk (probability of occlusion) in color-flow surveillance grafts is less than one third of the relative risk in bypass grafts that underwent clinical follow up. CONCLUSIONS: We conclude that revision procedures were more optimally used during color-flow surveillance, whereas asymptomatic stenotic graft lesions are missed with clinical follow-up, which results in a higher percentage of graft failures. Overall graft patency rates can be improved with use of color-flow duplex surveillance and repair of significant stenotic lesions. PMID- 8421342 TI - Does correction of stenoses identified with color duplex scanning improve infrainguinal graft patency? AB - PURPOSE: This study was undertaken (1) to determine whether correction of infrainguinal bypass stenoses detected with color duplex scanning (CDS) improved graft survival and (2) to define the natural history of grafts that did not undergo revision. METHODS: Over a 39-month period 462 color-flow duplex scans were obtained on 170 limbs with autogenous vein grafts. Grafts were scanned within 3 months of operation, at 6 and 12 months, and then yearly. Doubling of the velocity at any point in the graft-arterial system compared with the velocity immediately above or below (velocity ratio > or = 2.0) was the criterion adopted for identification of a hemodynamically significant (> or = 50%) diameter reduction. RESULTS: One hundred ten stenoses were detected in 62 (36%) of the limbs, of which 9 (8%) were in native vessels, 30 (27%) were at the anastomoses, and 71 (65%) were in the graft itself. Seventy-seven percent of the stenoses were detected in the first year. Twenty-four (39%) of the grafts with positive scans were revised. During follow-up, occlusions occurred in 10 (9%) of the 108 grafts with negative scans (NEG), in 2 (8%) of the 24 revised grafts with positive scans (PR), and in 10 (26%) of the 38 non-revised grafts with positive scans (PNR). Cumulative patency rates of NEG grafts were 90% at 1 year and 83% at 2 through 4 years. Similar patency rates were found in the PR vein grafts: 96% at 1 year and 88% at 2 through 4 years. In contrast, patency rates in PNR grafts with 50% or greater stenoses were only 66% at 1 year and 57% at 2 through 4 years. Log-rank tests showed a significant difference between the cumulative patency rates of NEG and PNR grafts (p < 0.002) and between PR and PNR grafts (p = 0.02). Flow velocities less than 45 cm/sec and ankle/brachial indexes did not discriminate well between grafts with or without 50% or greater stenoses or identify those grafts that subsequently occluded. CONCLUSIONS: The results of this study suggest that CDS detects graft-threatening lesions, that a velocity ratio of 2.0 or greater is the most highly predictive parameter, and that revision of grafts with stenoses identified with CDS prolongs patency. PMID- 8421343 TI - Surgical treatment of infrainguinal arterial occlusive disease in women. AB - PURPOSE: This study reviewed the outcome of 131 women who underwent infrainguinal bypass in 150 limbs from 1984 to 1991 for limb-threatening ischemia (95%) or disabling claudication (5%). METHODS: These women were compared with 209 men who underwent infrainguinal arterial reconstruction of 231 lower extremities for limb threat (89%) or claudication (11%) during the same interval. On average, women were 3 years older than men (mean age 72 vs 69 years, p < 0.005) but were less frequently cigarette smokers (56% women, 68% men, p < 0.05). Fifty-two percent of women had diabetes and 67% had hypertension, similar to the male patients. Infrainguinal disease distribution necessitated bypass to the above-knee popliteal artery in 10%, to the below-knee popliteal artery in 25%, and to the tibial or pedal arteries in 65% of women, comparable to the disease distribution in men. Autogenous vein grafts were performed in 90% of both groups. RESULTS: Early postoperative (30-day) mortality was 4% for women and 2% for men (not significant). Life-table survival after 3 years, however, was only 54% in women, compared with 72% in men (p < 0.05). Multivariate analysis indicated that diabetes increased the mortality rate 2.5-fold in women, which was not true in men. Three-year life-table survival of women with diabetes was only 39%, compared with 78% in women without diabetes (p < 0.001). Primary graft patency in women was 59% at 1 year and 54% at 3 years, significantly less than the 73% and 70% graft patency rates observed in men (p < 0.005). Secondary graft patency improved in women to 75% and 69% after 1 and 3 years, but this was still significantly less than the secondary patency rates of 89% and 86% observed in men (p < 0.001). Multivariate analysis indicated that female sex decreased secondary graft patency 2.4-fold and was the only variable associated with graft failure. Cumulative 3 year limb salvage in women was 82%, not statistically different than the 89% limb salvage rate observed in men. CONCLUSIONS: Women and men requiring arterial reconstruction for infrainguinal occlusive disease had comparable operative mortality and limb salvage rates, but long-term survival and graft patency were significantly reduced in women. Our results indicate that sex substantially influences the outcome of patients after infrainguinal bypass. PMID- 8421344 TI - John Hunter--triumph and tragedy. PMID- 8421345 TI - Mesenteric duplex scanning: a blinded prospective study. AB - PURPOSE: Based on retrospective comparisons of duplex scanning with arteriography of the celiac (CA) and superior mesenteric (SMA) arteries in 34 patients, we previously suggested that an SMA peak systolic velocity of 275 cm/sec or greater or no flow signal and a CA PSV of 200 cm/sec or greater or no flow signal were reliable indicators of a 70% or greater angiographic stenosis of the SMA and CA, respectively. We now report the results of a blinded, prospective study in a larger patient group designed to determine the ability of mesenteric duplex scanning to visualize the CA and SMA and to validate our proposed duplex criteria for splanchnic artery stenosis. METHODS: During an 18-month period 100 patients admitted to our vascular surgery service for aortography underwent routine mesenteric artery duplex scanning and lateral abdominal aortography regardless of abdominal symptoms. The lateral aortograms were evaluated to determine the presence or absence of a 70% or greater stenosis in the CA or SMA. Duplex determined peak systolic velocities from the CA and SMA were recorded without knowledge of the angiographic results. RESULTS: Aortography satisfactorily visualized 100% of the CAs and 99% of the SMAs. Of these, 93% of the SMAs and 83% of the CAs were visualized by duplex. According to the above criteria, duplex sensitivity, specificity, positive predictive value, negative predictive value, and overall accuracy for detection of a 70% or greater SMA stenosis were 92%, 96%, 80%, 99%, and 96% and for a 70% or greater CA stenosis 87%, 80%, 63%, 94%, and 82%. CONCLUSIONS: Mesenteric duplex scanning is feasible in the majority of patients. Prospective evaluation of duplex diagnostic criteria for 70% or greater stenosis indicates that mesenteric duplex scanning is sufficiently accurate to be clinically useful as a screening examination to detect SMA and CA stenosis. PMID- 8421346 TI - Detection of aortic graft infection with leukocytes labeled with technetium 99m hexametazime. AB - PURPOSE: To reduce the rates of morbidity and mortality in aortic graft infection, a new diagnostic approach is needed to help identify low-grade stages, specifically when there are minimal or no clinical signs of overt infection. The aim of this study was to evaluate the role of technetium 99m--hexametazime white blood cell scanning (99mTc scanning) in detecting aortic graft infection, particularly in the low-grade stages. METHODS AND RESULTS: Thirty-seven patients with suspected aortic graft infection were categorized into three groups according to their signs and symptoms on readmission. Ten patients (group A) had advanced graft infections that were correctly diagnosed by use of computed tomography (CT) scanning and 99mTc scanning and confirmed by intraoperative findings and culture results. Eighteen patients (group B) had nonspecific signs and symptoms of graft infection. Patients only underwent CT and 99mTc scanning for graft infection after standard clinical work-ups failed to reveal disease processes that accounted for the clinical symptoms. In this group of patients 99mTc scanning identified four cases of low-grade graft infection, which was confirmed by intraoperative findings and graft cultures. None of these four cases was confirmed by results of CT scanning. On an average 18-month follow-up in patients who did not undergo surgery graft infections developed. Nine patients (group C) had anastomotic aneurysms; CT scanning and 99mTc scanning correctly diagnosed five patients as being infected. The result of 99mTc scanning was false positive in one patient. CONCLUSIONS: The diagnostic accuracy of 99mTc scanning in patients who did not have specific signs of graft infection (groups B and C) was 100% for sensitivity, 94.4% for specificity, 90% for the positive predictive value, and 100% for the negative predictive value. 99mTc scanning seems to be a useful diagnostic technique for detecting aortic graft infection, particularly in low-grade stages. PMID- 8421347 TI - Determinants of successful peroneal artery bypass. AB - PURPOSE: The purpose of this study was to identify the determinants of long-term success with the peroneal artery bypass. METHODS: Seventy-seven consecutive peroneal artery bypasses performed between 1981 and 1990 were reviewed to evaluate the influence of venous conduit modification, surgeon experience, patency of the dorsalis pedis or posterior tibial artery at the ankle, and the quality of the peroneal collateral branches and pedal arch. The average follow-up was 34 months (range 1 to 92 months). RESULTS: The 5-year primary and secondary patency rates were 61% and 92%, respectively. Modification of the vein graft at the initial procedure was necessary in 26 (34%). This led to a reduced (p < 0.001) 5-year primary patency rate of 22% compared with 80% for unmodified conduits. Operative results improved with surgeon experience. The 5-year secondary patency rate of grafts placed before 1985 was 82% compared with 98% for subsequent grafts (p < 0.03). The initial postoperative mean ankle/brachial index for grafts revised for hemodynamic failure or thrombosis in the follow-up period was 0.84 compared with 0.95 in grafts that did not require revision (p < 0.04). The presence of a patent dorsalis pedis or posterior tibial artery at the ankle and an intact pedal arch did not significantly influence primary or secondary patency. The 5-year secondary graft patency rate for patients with a patent dorsalis pedis or posterior tibial artery at the ankle was 88%. CONCLUSIONS: The peroneal artery should be selected for outflow when it is the single tibial runoff vessel and is preferable to a bypass to an inframalleolar arterial segment. The quality of the venous conduit and the technical skill of the surgeon are the two most important factors in the success of bypasses to the peroneal artery. PMID- 8421348 TI - A novel, unstable DNA mutation cracks decades-old clinical enigma. PMID- 8421350 TI - Military medicine prepares health and safety tips for those serving in Somalia, requests feedback. PMID- 8421349 TI - Triplet repeat mutations: amplification within pedigrees generates three human diseases. PMID- 8421351 TI - Medicare case underlines importance of physician compliance with all rules when claims are filed. PMID- 8421352 TI - From the National Institutes of Health. PMID- 8421353 TI - From the Centers for Disease Control and Prevention. Acute respiratory illness linked to use of aerosol leather conditioner--Oregon, 1992. PMID- 8421354 TI - From the Centers for Disease Control. Deaths and hospitalizations from chronic liver disease and cirrhosis--United States, 1980-1989. PMID- 8421355 TI - From the Centers for Disease Control. Air pollution information activities at state and local agencies--United States, 1992. PMID- 8421356 TI - The definition of alcoholism. PMID- 8421357 TI - High-altitude syncope: history repeats itself. PMID- 8421358 TI - Waived and nonwaived tests: the Clinical Laboratory Improvement Act. PMID- 8421359 TI - Excision margins for melanoma in awkward places like the eyelid. PMID- 8421360 TI - Hepatitis B in a prenatal population. PMID- 8421361 TI - Q-wave vs non-Q-wave infarction: an oversimplified dichotomy. PMID- 8421362 TI - Euthanasia: should the public decide? PMID- 8421363 TI - Moderate caffeine use and the risk of spontaneous abortion and intrauterine growth retardation. AB - OBJECTIVE: To examine the relationship between caffeine consumption during pregnancy and the occurrence of spontaneous abortion and intrauterine growth retardation. DESIGN, SETTING, AND PATIENTS: A cohort of 431 women, enrolled in a multicenter study within 21 days of conception, was monitored throughout pregnancy to determine (1) caffeine exposure, (2) exposure to other risk factors, (3) fetal growth as assessed by ultrasonography, and (4) pregnancy outcome. OUTCOME MEASURES: Spontaneous abortion, intrauterine growth, birth weight, and head circumference. RESULTS: The mean (+/- SD) first-trimester caffeine consumption was not significantly higher in women who aborted (125.9 +/- 123.1 mg) than in women who delivered liveborn infants (111.6 +/- 107.0 mg) (P = 34). The adjusted odds ratio (OR) for spontaneous abortion was 1.15 (95% confidence interval [CI], 0.89 to 1.49). Early fetal growth, assessed by crown-rump length on ultrasonographic examination, was not affected by caffeine. Although the group consuming the most caffeine (> 300 mg/d) had a significantly higher proportion of babies with birth weights and head circumferences below the 10th percentile in the crude analysis, the association with caffeine was no longer significant when other risk factors (notably smoking) were taken into account. The adjusted ORs were 1.11 (95% CI, 0.88 to 1.40) for decreased birth weight and 1.09 (95% CI, 0.86 to 1.37) for smaller head circumference. CONCLUSIONS: Close monitoring of a cohort identified very soon after conception enabled us to identify all abortions after 21 days postconception, monitor intrauterine growth prospectively, and track caffeine use. Despite this intensive surveillance, we found no evidence that moderate caffeine use increased the risk of spontaneous abortion, intrauterine growth retardation, or microcephaly after accounting for other risk factors. PMID- 8421364 TI - The impact of comorbid and sociodemographic factors on access to renal transplantation. AB - OBJECTIVE: To assess the impact of sociodemographic factors and comorbid conditions on access to renal transplantation for adult US dialysis patients with end-stage renal disease (ESRD). DESIGN: Cohort analytic study. Data on comorbid conditions at onset of ESRD were abstracted from patients' medical records and matched to sociodemographic and ESRD data from the United States Renal Data System database. SETTING: United States Medicare dialysis population. PATIENTS: Random, national sample of ESRD patients starting dialysis in 1986 and 1987 (n = 4118). MAIN OUTCOME MEASURE: Time to first renal transplant (living or cadaver donor) since onset of ESRD regressed with two nested Cox proportional hazards models, first against sociodemographic factors alone, and then against sociodemographic factors and comorbid conditions. RESULTS: Cardiovascular diseases are most predictive of who received a transplant; patients with coronary heart disease, congestive heart failure, or left ventricular hypertrophy showed lower transplantation rates relative to patients without the disease (relative rate [RR] = 0.65 to 0.80, P < .05 each). Obese patients and patients with peripheral vascular disease also showed lower transplantation rates (RR = 0.65 to 0.75, P < .05 each). Previously reported sociodemographic effects of lower transplantation rates for older patients, women, nonwhite patients, and lower income patients were confirmed (P < .01). Sociodemographic effects remained essentially unchanged when adjusted for comorbid conditions. CONCLUSIONS: These findings indicate that sociodemographics have strong independent effects on access to transplantation that cannot be explained away as "surrogate" effects related to comorbid factors. Furthermore, the results suggest that lower mortality rates for transplant recipients relative to dialysis patients are due, in part, to a healthier case mix among patients receiving transplants. PMID- 8421365 TI - Influence of diagnostic criteria on the incidence of gestational diabetes and perinatal morbidity. AB - OBJECTIVE: To determine the incidence of gestational diabetes and its associated maternal and infant morbidity by two sets of 3-hour glucose tolerance test criteria, those recommended by the National Diabetes Data Group or the lower, modified criteria of Carpenter and Coustan. DESIGN: Prospective, observational outcome cohort study. SETTING: Prepaid health maintenance organization. SUBJECTS: A total of 2019 pregnant women preregistered in a health maintenance organization were screened for a plasma glucose of 7.77 mmol/L (140 mg/dL) or greater, 1 hour after ingestion of a 50-g glucose drink administered after an overnight fast. Positive subjects received a 3-hour glucose tolerance test interpreted by the two criteria. Data are presented for 521 randomly selected negative screenees; 264 positive screen, negative glucose tolerance test subjects; and 101 subjects with gestational diabetes mellitus. OUTCOME MEASURES: Maternal risk factors for diabetes, infant birth weight corrected for gestational age (birth-weight ratio), umbilical cord serum insulin levels, and 33 maternal and infant perinatal morbidities assessed from chart review. RESULTS: Gestational diabetes incidence was 5.0% overall based on the modified criteria and 3.2% by the recommended criteria. Maternal age and family history of diabetes were higher in both modified and recommended groups. Birthweight ratio was 1.05 in negative screenees, 1.09 in gestational diabetes overall (P < .05 when only diet-treated subjects were considered), and 1.11 in modified and 1.08 in recommended criteria groups. Cord serum insulin levels in infants of gestational diabetic mothers by both criteria were 40% above those of negative screenees (P < .001). The average percentage incidence of 33 possible perinatal morbidities was 41% higher in gestational diabetic pregnancies by the modified criteria. The cumulative number of morbidities was higher in both modified and recommended criteria groups (P < .01 in both instances). In contrast, subjects having a positive glucose screening test but a negative glucose tolerance test by modified criteria had none of the characteristics of gestational diabetes with the single exception of greater age. CONCLUSIONS: Fifty percent more cases of gestational diabetes were identified using the more inclusive, modified criteria. These cases had as much excess in maternal diabetes risk factors, infant macrosomia, and cord hyperinsulinemia and nearly as much increase in perinatal morbidity as subjects diagnosed by the recommended criteria. The incidence and perinatal impact of gestational diabetes may be greater than previously appreciated. The modified criteria deserve wider verification and use. PMID- 8421366 TI - Preventive medicine for our ailing health care system. PMID- 8421367 TI - Diabetic ketoacidosis in prisoners without access to insulin. AB - OBJECTIVE: To assess the cause and clinical severity of diabetic ketoacidosis in male prisoners hospitalized in New York City. DESIGN: Retrospective chart review. SETTING: A municipal hospital in New York City. PATIENTS: Forty-nine adult male prisoners with a total of 54 hospital admissions for diabetic ketoacidosis between January 1, 1989, and June 30, 1991. MAIN OUTCOME MEASURES: Charts were reviewed for diabetic and medical history, time from arrest until hospitalization, cause of diabetic ketoacidosis, admission laboratory data, and hospital course. RESULTS: Thirty-eight (70%) of the 54 admissions for diabetic ketoacidosis among prisoners occurred because prisoners had not received insulin during the period immediately following arrest (mean number of days from arrest until hospitalization was 2.5). All of these individuals had a history of insulin dependent diabetes and were reportedly compliant with their insulin regimen at the time of arrest. Admission laboratory data for this group of prisoners included a mean serum glucose level of 27.4 mmol/L (495 mg/dL) and a mean serum bicarbonate level of 14.4 mmol/L. Mean number of days in the hospital was 3.4 including a mean of 1 day in an intensive care unit. CONCLUSIONS: Inadequate access to medication results in serious sequelae for recently arrested prisoners in New York City with insulin-dependent diabetes. Access to health care for recently arrested prisoners needs to be improved. PMID- 8421368 TI - No pain, no gain. Perspectives on cost containment. PMID- 8421369 TI - Health care for prisoners. How soon is soon enough? PMID- 8421370 TI - A piece of my mind. No shattered vision. PMID- 8421371 TI - America's aging population: changing the face of health care. PMID- 8421372 TI - Aspects of the medical history unique to older persons. PMID- 8421373 TI - [Methods of economizing donor blood--the legal situation]. PMID- 8421374 TI - [Risks of homologous blood transfusion]. PMID- 8421375 TI - [Donor blood conserving surgical techniques in acute traumatology]. PMID- 8421376 TI - [Preoperative autologous blood donation]. PMID- 8421377 TI - [Autologous plasmapheresis]. PMID- 8421378 TI - [Hemodilution]. PMID- 8421379 TI - [Mechanical autotransfusion]. PMID- 8421380 TI - [Methods p6 economizing donor blood. Appendix. Recommendations for the adoption of methods of economizing donor blood]. PMID- 8421381 TI - [Risks of autologous blood transfusion]. PMID- 8421382 TI - [The critical hematocrit--an analysis]. PMID- 8421384 TI - [Artificial and biological volume replacement solutions]. PMID- 8421383 TI - [Cardiovascular risks of "methods of economizing donor blood"]. PMID- 8421385 TI - [Hemostatic disorders in operative medicine: the place of treatment with blood components and the effect of the use of artificial volume replacement solutions]. PMID- 8421386 TI - [Indications, effectiveness and limitations of controlled hypotension]. PMID- 8421387 TI - [Blood saving operations in abdominal surgery]. PMID- 8421388 TI - Andreas Vesalius on the larynx and hyoid bone: an annotated translation from the 1543 and 1555 editions of De humani corporis fabrica. PMID- 8421389 TI - Caesarean section in the Maltese Islands. PMID- 8421390 TI - The development of ethical guidance for medical practitioners by the General Medical Council. PMID- 8421391 TI - The miswak, an aspect of dental care in Islam. PMID- 8421392 TI - Illustrations from the Wellcome Institute Library. Some recently acquired papers of Robert Whytt (1714-1766), FRS. PMID- 8421393 TI - [Time for continuing education of "surgeons for adults?" Pediatric surgery is neglected in the county health service]. PMID- 8421394 TI - [The barons von Munchhausen were not all liars and cheats!]. PMID- 8421395 TI - [Save time and money--let the patient decide!]. PMID- 8421396 TI - [Conservatism prevents widening of psychotherapeutic education]. PMID- 8421397 TI - [Preparations for smoking cessation to pregnant women?]. PMID- 8421398 TI - [A revolution in Linkoping. The patient in the centre]. PMID- 8421399 TI - [Is Swedish emergency medicine better than the English? Cost savings a threat to quality]. PMID- 8421400 TI - [Myocardial hypertrophy and fibrosis. New connections with the renin-angiotensin aldosterone system]. PMID- 8421401 TI - [Acute fulminant liver failure. Importance of early contact with a specialist]. PMID- 8421402 TI - [Migraine related stroke. New classification makes for better research]. PMID- 8421403 TI - [Passive smoking. Limited risk factor of fetal damage]. PMID- 8421404 TI - [Mycobacterium infections. New techniques are going to revolutionize diagnosis]. PMID- 8421405 TI - [Quincke's edema and photosensitization in a new report of adverse effects]. PMID- 8421406 TI - [What causes macular degeneration? 2 recently identified components can be the answer]. PMID- 8421407 TI - [Basic diet in Crohn disease. Good complement to drugs for children and adolescents]. PMID- 8421408 TI - [Adder bite caused severe thrombocytopenia]. PMID- 8421409 TI - [Living wills law in Denmark. Rapid input of 25,000 wills to a nation-wide registry]. PMID- 8421410 TI - Functional endonasal sinus surgery in adults and children. AB - Functional endonasal sinus surgery (FESS) is becoming the procedure of choice for the surgical treatment of chronic and recurrent sinusitis in adults and children. Retrospective analysis of the charts of 513 adult and 260 pediatric patients who underwent FESS after failing to respond to optimal medical treatment revealed an improvement rate of approximately 80% for both age groups. Although high response rates and low complication rates were found for both groups, there were significant differences in indications, preoperative evaluation, operative technique, and methods of postoperative follow-up for children. PMID- 8421411 TI - Excision of rhinophyma with Nd:YAG laser: a new technique. PMID- 8421412 TI - Directory of otolaryngological societies. PMID- 8421413 TI - Serologic determinants of survival in patients with head and neck cancer: validating a clinical prediction model. AB - Quantitative measurements of serum C1q-binding macromolecules (C1qBM) and immunoglobulin A (IgA) were done on 162 patients using previously described methodology. The measurements were compared to a previously described head and neck cancer population. Using the Cox Proportional Hazards model, the prognostic implications regarding high C1qBM and subsequent death with disease (P = .02), and regional recurrence (P = .0094) were validated, but not our previous IgA related prognostic implications. When both study populations were combined, C1qBM was predictive of survival in those patients treated with induction chemotherapy (P = .0001). C1qBM was not a significant predictor of survival in patients treated with surgery plus postoperative radiation therapy in either this second "test" population or in the original "training" population. The findings demonstrate the confounding influence of treatment modalities and the importance of model validation. PMID- 8421414 TI - Lymphoma of the parotid gland. AB - In a consecutive group of 452 patients undergoing parotid surgery at this institution, 18 (4%) were found to have lymphoma. Review and analysis of presenting symptoms, predisposing factors, histopathology, postsurgical morbidity, and long-term outcome with treatment are presented. The current literature on parotid lymphoma is reviewed, and management strategies are outlined. Although a relatively uncommon primary lesion, lymphoma must be considered in the differential diagnosis of any mass presenting in the parotid gland. PMID- 8421415 TI - Nonwound infections following head and neck oncologic surgery. AB - Little information exists regarding the comorbidity of postoperative nonwound infections (NWIs) in patients with head and neck cancer. Prospectively, 225 patients were randomized in a double-blind fashion to receive either clindamycin or ampicillin sodium/sulbactam sodium for prevention of postoperative wound infection. Of the 113 patients receiving clindamycin, 14 developed nonwound infections, compared with 10 of 112 patients receiving ampicillin/sulbactam. A single site of nonwound infection occurred in 21 patients, and 2 sites occurred in 3 patients. The majority of infections were pulmonary (22), followed by urinary tract (3), septic phlebitis (1), and acute sinusitis (1). Gram-negative organisms were isolated more frequently among patients on clindamycin (18) versus ampicillin/sulbactam (6) (P = .014). Risk factors for pulmonary nonwound infection included: longer surgery, a greater than 70 packs per year smoking history, blood transfusion, and hypoalbuminemia (P < .05). Nonwound infections produce significant postoperative morbidity and the predominance of gram-negative organisms isolated from these infections has therapeutic implications. PMID- 8421416 TI - Subperiosteal orbital abscess in children: diagnosis, microbiology, and management. AB - A chart review was undertaken of 30 patients who were presented to Children's Hospital of Pittsburgh from 1983 to 1990 and underwent surgical management for suspected subperiosteal abscess of the orbit (SPA). All patients had preoperative computerized tomography (CT) scans. Twenty-seven of 30 CT scans were reevaluated, of which 20 (74%) demonstrated findings consistent with or suggestive of SPA. Opacified sinuses were seen in 100% of this study group. Abscess collections were found in 21 patients (70%). Predominant organisms included: Streptococcus pneumoniae (38%), Streptococcus pyogenes (33%), and Haemophilus influenzae (14%); Moraxella catarrhalis was not isolated and anaerobic bacteria were isolated from only one patient. CT scans were found to be accurate predictors of SPA in 16 (80%) of 20 patients. All patients did well following surgical intervention without visual or central nervous system sequelae. We conclude from this study that CT scans should be obtained when SPA is suspected, and antimicrobial therapy should be directed toward the bacteria isolated from these abscesses. PMID- 8421417 TI - Once-a-day therapy for sinusitis: a comparison study of cefixime and amoxicillin. AB - The efficacy and safety of a once-a-day antibiotic in the treatment of sinusitis was studied. Two randomly assigned groups were treated with either once-a-day cefixime, a third generation cephalosporin, or amoxicillin three times a day. One hundred and fourteen patients were evaluated with antral punctures, microbiologic evaluation, and radiographic studies. Cultures revealed 40% gram-negative organisms, 48% gram-positive, and 12% anaerobes. The most common bacteria were Haemophilus influenzae, Streptococcus pneumoniae, Staphylococcus aureus and viridans group streptococci. Ninety-four percent of the cefixime group were cured compared with 96% of the amoxicillin group. Staphylococcus resistance was a problem in both groups, necessitating an occasional change to amoxicillin clavulanate potassium in the amoxicillin group. Once-a-day antibiotics offer the potential for improved compliance in the treatment of sinusitis. Cefixime offers an additional benefit of covering beta-lactamase producing strains of bacteria which are increasing in incidence and resistant to many penicillins. PMID- 8421418 TI - Oncogenes in head and neck cancer. AB - Primary head and neck squamous cell carcinomas from 58 patients were analyzed for the presence of alterations in the K-ras, raf, and erb-B oncogenes. Analysis of 17 fresh tumor specimens using sequence analysis of target sequences amplified by the polymerase chain reaction showed no evidence of mutations in the K-ras oncogene. Thirty fresh tumor specimens were analyzed for the presence of raf gene activation using Southern blot analysis. Under these conditions, no mutations in the c-raf oncogene were detected. In this study, 6 (13%) of the 47 tumors studied displayed epidermal growth factor receptor (EGFR) gene amplification and/or gene rearrangement. Overexpression of the erb-B gene was observed in 18 (67%) of the 27 head and neck tumors studied. Those patients expressing high levels of EGFR or showing EGFR amplification had tumors that were clinically more advanced. These data suggest that amplification and over-expression of the EGFR gene may be a useful diagnostic and prognostic marker in head and neck squamous cell carcinomas. PMID- 8421419 TI - Blastomycosis in otolaryngology: review of a large series. AB - Blastomycosis is a fungal disease whose signs, symptoms, and roentgenographic findings may suggest pneumonia, cancer, tuberculosis, or other fungal infections. Blastomycosis has a variable clinical course; it primarily affects the lower respiratory tract, although extrapulmonary manifestations are common. To determine the frequency of head and neck manifestations, we reviewed the records of patients with blastomycosis seen at the Mayo Clinic from 1960 to 1990 and found that involvement of the skin and mucosal surfaces, including the larynx, was common. Gross and histopathologic features of the lesions often resemble those of well-differentiated squamous cell carcinoma; thus, overly aggressive therapy may be mistakenly instituted. The clinical features, morphologic findings, and diagnosis of blastomycosis are discussed, and the importance of obtaining and handling specimens correctly is emphasized. PMID- 8421420 TI - Neck needle foreign bodies in intravenous drug abusers. AB - Intravenous drug abusers often resort to deep cervical venous access if peripheral access is problematic. Deep cervical injection can occasionally result in needle breakage and needle foreign body. The records of 50 patients with neck needle foreign bodies related to intravenous drug abuse treated at The Johns Hopkins Hospital were reviewed retrospectively. Only half of these patients presented to this hospital with a complaint of a neck needle. Complications related to neck needles were manifested in 5 (10%) of the patients at the time of initial presentation. No delayed complications occurred. Human immunodeficiency virus seropositivity for this group of patients was 77% compared to a rate of 29% for all intravenous drug abusers at the same institution. The demographics and management of neck needle foreign bodies are discussed, and the importance of recognition of neck needle foreign bodies is emphasized. PMID- 8421421 TI - Effect of a topical vasoconstrictor on computed tomography of paranasal sinus disease. AB - Apparent ethmoid inflammation which resolved with alternation of the nasal cycle or following application of topical vasoconstrictors has been observed with magnetic resonance imaging. A similar phenomenon might occur to a lesser degree with computed tomography (CT), leading to overdiagnosis of limited sinus disease. The degree to which ostiomeatal complex disease is reversible by topical vasoconstrictors was investigated. Ten patients with histories of chronic or recurrent sinusitis underwent coronal CT studies of the paranasal sinuses before and after the application of a topical vasoconstrictor. Mucosal volume or thickness measurements were obtained from the turbinates, infundibulum, ethmoidal cells, and antrum. Vasoconstrictor application markedly reduced turbinate size and appeared to reduce mucosal thickening in the ethmoidal infundibulum. Minimal mucosal changes identified in the paranasal sinuses by computed tomography were not reversed by vasoconstrictors and therefore are likely to be pathologic. PMID- 8421422 TI - Implications of positive surgical margins. AB - The recently concluded Head and Neck Intergroup trial tested the addition of three courses of cis-platinum containing chemotherapy to standard treatment of surgery and postoperative radiotherapy for patients with advanced operable squamous cell carcinoma of the head and neck. Only patients with negative surgical margins were eligible for the trial. One hundred twelve patients with positive surgical margins were dropped from the trial but continued to be followed. These patients received a variety of treatments. Positive surgical margins were most often seen in nonglottic primaries and with increasing frequency as the N stage increased. Patients with positive margins who achieved a complete clinical response to subsequent treatment had a median survival of 33.8 months vs. 9.1 months for those with less than a complete clinical response. The addition of chemotherapy did not significantly alter the median survival of the positive margin patients. PMID- 8421423 TI - Clinical staging for primary malignancies of the supraglottic larynx. AB - A study of 520 patients with primary supraglottic cancer was conducted. The tumors were staged according to the 1983 and 1988 American Joint Committee on Cancer (AJCC) T- and N-stage definitions. There were 293 patients with early stage (T1, T2) tumors, 227 with advanced stage (T3, T4) tumors and 428 with early nodal disease (N0, N1) in both systems. In the 1983 N-staging, there were 44 N2 and 48 N3 lesions; in the 1988 N-staging, there were 62 N2 and 30 N3 lesions. Cox regression analysis showed that the 1983 and 1988 T-stage (T1 through T4) definitions successfully prognosticate for survival when patients were without neck node involvement. In contrast, when neck nodes were present, the N-stage (N0 through N3) of the disease prognosticated survival better than T-stage. Further analyses showed that the 1988 N-stage definition provided a better separation between N2 and N3 lesions compared to the 1983 version. Combined-modality treatment (surgery and radiation) significantly improved survival compared to single-modality treatment (surgery or radiation alone) when patients were staged T4 and N0 through N3 neck disease, but not when patients advanced from T1 to T3. Comparison of treatment efficacy over the last four decades for single- and combined-modality treatment did not reveal statistically significant differences in survival rates in our patient population. This was consistent with cumulative results of various institutions over the last four decades. We conclude that the 1988 AJCC T- and N-stage definitions successfully prognosticate for T-stage (T1 through T4) and N-stage (N0 through N3) with better separation of N2 and N3 lesions compared to the 1983 version. PMID- 8421424 TI - Bony dehiscence between singular canal and round window niche. AB - The incidence of bony dehiscence of the human singular canal was studied by examining 409 human temporal bones obtained from 300 individuals. Such a dehiscence was noted in three (0.7%) of the bones from three (1%) of the individuals, all male, aged 2, 6, and 76 years. All of these bony dehiscences were located between the round window niche and the cribrose area of the singular canal. The finding of a microfissure in the vicinity of the bony dehiscence in two of the three cases indicates that such dehiscences occur independently of microfissures. Although the incidence of bony dehiscences of the singular canal is small, these dehiscences appear to have considerable clinical significance in that they form a communication between the middle and inner ears or possibly between the middle ear and the cerebrospinal fluid space. PMID- 8421425 TI - Hemipharyngectomy and hemilaryngectomy for pyriform sinus cancer: reconstruction with remaining larynx and hypopharynx and with tracheostomy. AB - When resection of the posterior hypopharyngeal wall is undertaken for treatment of pyriform sinus carcinoma, a very narrow mucosal strip may be left for reconstruction. A surgical technique for carcinoma which has invaded the whole lateral hypopharyngeal wall is described. It consists of resecting half the larynx and half the hypopharynx and reconstructing the food canal using the mucosa of the unaffected half of the larynx and the remnant hypopharynx. A series of 34 patients treated by this technique is presented. No hypopharyngeal stenosis was observed. Local recurrences developed in 9 cases and distant metastases in 6. A specific disease mortality rate of 44% was encountered. At present, 15 patients are disease-free after a median follow-up of 48 months. PMID- 8421426 TI - Peripheral hearing loss following head trauma in children. AB - The purpose of this prospective study was to determine the incidence and type of hearing loss occurring in children who suffered head injuries. Fifty children admitted to the neurosurgical service after sustaining head trauma were studied. Neurologic, otologic, and audiologic evaluations were performed. Diagnostic studies included skull roentgenograms and computerized tomography scans. A 32% incidence of conductive hearing loss and a 16% incidence of high-frequency sensorineural hearing loss was found in this group. All patients with temporal bone fractures had conductive hearing losses, but the presence of a skull vault fracture did not correlate with the presence, type, or degree of hearing loss. In addition, there was no correlation between either cause of injury, loss of consciousness, or Glasgow Coma Scale scores and the presence, type, or degree of hearing loss. There was a significant incidence of both sensorineural and conductive hearing loss in this series of patients, which indicates that close audiologic and otologic follow-up is necessary for all head injury patients. PMID- 8421427 TI - A safe, effective anesthetic technique for outpatient myringotomy tube placement. PMID- 8421428 TI - Composite TORPs and PORPs for ossicular reconstruction. PMID- 8421429 TI - Free jejunal autografts for immediate voice restoration following laryngectomy. PMID- 8421430 TI - Anti-allergic activity of roxithromycin: inhibition of interleukin-5 production from mouse T lymphocytes. AB - This study was designed to evaluate the effects of roxithromycin (RXM), a newly synthesized macrolide antibiotic on allergic responses in mice. RXM was orally administered into BALB/c mice once a day for 42 days in a single dose of 5 mg/kg body weight. Spleen cells (Sp C) collected from mice on day 7, 14, 28 and 42 post RXM administration showed higher blastic activity of lymphocytes than those from control. The activity peaked on the 7th day, then gradually decreased, and returned to the control level by the 42nd day. Production of cytokines, IL-2 and IL-5, by Sp C in response to concanavalin A stimulation was also examined in the course of RXM administration. The capacity of Sp C to produce IL-2 was enhanced by oral administration of RXM for 28 days. However, a long-term (for 42 days) administration inhibited it. On the other hand, the capacity of of Sp C to produce IL-5 was strongly inhibited by oral administration of RXM; the titer of IL-5 was similar to that obtained in cultures of Sp C from control mice. These results strongly suggest that oral administration of RXM inhibits the function of Th2-type helper T lymphocytes and that a long-term administration of RXM may be beneficial in asthma and allergy. PMID- 8421431 TI - Endothelial potentiation of relaxation response to ascorbic acid in rat and guinea pig thoracic aorta. AB - The role of the endothelium was evaluated in the relaxation of rat and guinea pig aortic rings induced by ascorbic acid. Ascorbic acid relaxed rat and guinea pig aortic rings that were previously contracted with submaximal dose of phenylephrine (PE), in a concentration dependent manner. Removal of the endothelium significantly reduced the sensitivity but not the magnitude of the response to ascorbic acid. Methylene blue, but not propranolol, blocked the endothelial augmentation of vascular relaxation to ascorbic acid. Vessels precontracted with potassium chloride (high K+) were also relaxed by ascorbic acid. Methylene blue also inhibited the response to ascorbic acid in the intact vessels precontracted with high K+. A23187 and acetylcholine, but not ADP, variably caused endothelium dependent component relaxation in guinea pigs, whereas all of these three probes constantly caused it. In Ca(2+)-free medium, Ca(2+)-induced contraction of high K(+)-depolarized rat aorta was inhibited by the presence of ascorbate, which was more pronounced in endothelium intact rings than in endothelium denuded ones. PE-induced contraction in the presence of different concentrations of ascorbate reduced both the sensitivity and the maximal contractile force in rat aorta. Ascorbic acid (0.125-32 mM) did not change the pH in the medium. From these findings, it is speculated that 1) receptor- and potential-operated Ca2+ channels may be modulated by ascorbate, 2) endothelium has a significant role in promoting relaxation induced by ascorbic acid. PMID- 8421432 TI - Effects of in vivo morphine treatment on antibody responses in C57BL/6 bgJ/bgJ (beige) mice. AB - C57BL/6J bgJ/bgJ (beige) mice are less sensitive than other strains to the analgesic effects of morphine, although they have normal numbers of mu receptors. In the present study, beige mice and their normal littermates (beige+) were treated in vivo with morphine or the opioid antagonist, naltrexone and their primary in vitro antibody responses were assessed. Morphine treatment caused splenic atrophy and suppressed the primary in vitro antibody response in beige and beige+ mice. However, these effects were not blocked by naltrexone co treatment. In these mouse strains, naltrexone decreased spleen size and antibody responses by itself, which may mask its ability to antagonize morphine. In beige mice, placebo pellet implantation suppressed the primary in vitro antibody response. Morphine did not cause a further suppression of the antibody response in beige mice compared to placebo. Because of this anomalous response to placebo treatment, the immunosuppressive effects of morphine on the antibody response/10(7) cells can not be attributed to a specific drug effect in this strain. However, when antibody responses were expressed on a per spleen basis, the overall capacity to respond to antigenic challenge was suppressed by morphine treatment. PMID- 8421433 TI - Role of vanadium in nutrition: metabolism, essentiality and dietary considerations. AB - Vanadium is a pervasive element of biological systems, being widely distributed across the food supply. Food refining and processing appear to increase vanadium content. At higher intakes, it accumulates in body tissues such as liver, kidney and bone. Essentiality of the nutrient has been established in lower life forms but the significance and extent of vanadium's role in humans has been overshadowed by the absence of deficiency symptoms in man. While the pharmacological properties of vanadium have stimulated much interest, knowledge of basic metabolic processes regulating vanadium remains incomplete. Ultimate determination of essentiality for humans will depend on greater understanding of the fundamental biochemical roles of vanadium. PMID- 8421434 TI - Replacement of estrogen by methoxychlor in the artificially-induced decidual cell response in the rat. AB - The pesticide methoxychlor (MXC) exhibits estrogenic activity although it is not a steroid. Therefore its mode of action may differ from that of estrogen. Here we evaluated the ability of MXC to replace estrogen in the ovariectomized, hormone treated decidual cell response (DCR) model. Following priming with estrone, ovariectomized rats were treated with estrone plus progesterone, progesterone alone, or progesterone plus various dosage levels of MXC. Within a narrow dose range, MXC can replace estrone and, in combination with progesterone, produce a maximal DCR. In the same manner as that seen with progesterone plus estrone, progesterone plus MXC produced no effect different from progesterone alone at low to intermediate dosages and an inhibition of decidual growth at high doses. The data support the hypothesis that MXC exhibits classical mechanisms of estrogenic activity. PMID- 8421435 TI - Total parenteral nutrition-associated cholestasis after selective damage to acinar zone 3 hepatocytes by bromobenzene in the rat. AB - Total parenteral nutrition is known to cause cholestasis, but the hepatic site of this effect has not been determined. The purpose of our study was to observe the effect of TPN on bile flow and bile salt secretion rate in rats after selective damage to acinar zone 3. Bromobenzene, 3.8 mmol/kg, was injected i.p., and the animals were studied 48 hours later. Experimental groups received either parenteral nutrition or saline for 2 hours. Bromobenzene caused selective damage to acinar zone 3 hepatocytes, and reduced baseline bile flow (23.99 +/- 1.09 vs 37.2 +/- 1.66, mean +/- SEM, microliter/min/kg, p < 0.001). Bromobenzene had no effect on bile salt secretion rate. Total parenteral nutrition decreased bile flow in the bromobenzene treated groups, despite the selective hepatic damage to acinar zone 3 (20.54 +/- 1.07 vs 23.28 +/- 1.63, mean +/- SEM, p < 0.001). Total parenteral nutrition reduced bile salt secretion rate in healthy animals, but this reduction was not seen in bromobenzene treated rats. Our results suggest that bile flow reduction in response to total parenteral nutrition is mediated through an effect on acinar zones 1 and 2, as this reduction is still observed after zone 3 destruction by bromobenzene. Zone 3 hepatocytes may be involved in the effect of parenteral nutrition on bile salt secretion, as the reduction in secretion rate seen in healthy animals was not observed in bromobenzene treated rats. PMID- 8421436 TI - Endothelium-dependent contraction produced by acetylcholine and relaxation produced by histamine in monkey basilar arteries. AB - The present experiments were carried out to investigate endothelium-dependence of the responses to ACh, arachidonic acid and histamine in monkey basilar arteries. As we reported previously (1), ACh (10(-7) M) and arachidonic acid (5 x 10(-7) M) caused endothelium-dependent contraction (EDC) in both monkey and canine basilar arteries. The endothelium-derived contracting factor (EDCF) is probably TXA2, since the EDC was attenuated by a cyclooxygenase inhibitor (aspirin, 5 x 10(-5) M), thromboxane A2 (TXA2) synthetase inhibitors (OKY-046, 10(-5) M; RS-5186, 10( 6) M) and TXA2 antagonists (ONO-3708, 10(-8) M; S-145, 5 x 10(-9) M). Histamine caused endothelium-dependent relaxation (EDR) in monkey basilar arteries and EDC in canine basilar arteries. The EDR in monkey basilar arteries was attenuated by nitric oxide synthetase inhibitor, NG-monomethyl-L-arginine (L-NMMA) in a concentration-dependent manner and the EDC in canine basilar arteries was attenuated by aspirin, TAX2 synthetase inhibitors and TXA2 antagonists. The EDR and EDC were antagonized by tripelennamine (10(-6) M) but not by cimetidine (10( 5) M), indicating that they are mediated by H1-receptors. From these results, we suggest that in the monkey basilar artery, either there are two types of endothelium, an EDCF type for ACh and arachidonic acid and an EDRF type for histamine, or there is single type of endothelium with two types of signalling processes, one for EDC and one for EDR. PMID- 8421437 TI - [Analysis of early, late-phase and delayed skin reactions against 10 groups of antigens in 420 patients with allergic asthma]. AB - Immediate, late phase and delayed skin reactions against ten group of allergens were investigated in 420 patients with allergic asthma. The wheal and flare reaction were characteristic for type I IgE-mediated hypersensitivity reaction in human skin. This reaction develops rapidly after injection of antigen, peaks in 10-30 minutes, and then subsides within a few hours. However, careful observation for longer periods has shown that in many instances a late inflammatory response also appears at the same site and is quite different in appearance from the initial reaction. This late phase reaction sometimes lasts for 24 hours and it is characterized by a dense cellular infiltrate and is more oedematous than the early reaction. These late phase reactions can also be seen following challenge of the nasal mucosa and bronchi of allergic subjects and may be of major importance in the development of chronic asthma. In this study, we have investigated immediate, late phase also delayed phase skin reactivity incidence of the allergic asthma patients. PMID- 8421438 TI - [Determination of bacterial antigens by latex agglutination tests in cultured cerebrospinal fluid specimens from bacterial meningitis]. AB - Between December 1988 and February 1991, the latex agglutination test was evaluated for its ability to detect the antigens of N. meningitidis, S. pneumoniae and H. influenzae in the samples of cerebrospinal fluid (CSF) of 57 patients with bacterial meningitis. Totally antigens were detected in 34 of 42 (80.9%) patients with bacterial meningitis proven by culture. Latex agglutination was positive in 60% of CSFs for N. meningitidis, 88.2% of CSFs for S. pneumoniae and 100% of CSFs for H. influenzae. Antigens were detected in 7 of 9 (77.8%) patients with no bacterial organisms grown on CSF culture but seen on microscopy after Gram staining. No antigens were identified in CSFs of 4 patients with culture and gram stain negative. 32-48 hours after the treatment started, the CSF samples were obtained again from 11 patients and antigens were detected in 9 of them. PMID- 8421439 TI - [The antibacterial activity of the amniotic fluid during the last week of pregnancy]. AB - Amnion fluid samples were obtained from 100 pregnant women in the last week of pregnancy just before in labor. In detecting of antibacterial effectiveness, S. aureus ATCC 29213, E. coli ATCC 25922 and S. agalactiae ATCC 13813 strains were used. Antibacterial effectiveness of amnion fluid to these bacteria are found respectively as; 74%, 63% and 73.9%. PMID- 8421440 TI - [In vitro activity of ofloxacin and ciprofloxacin against Mycobacterium tuberculosis strains]. AB - The activity of Ofloxacin and Ciprofloxacin against 50 strains of M. tuberculosis were investigated in-vitro on Lowenstein-Jensen medium. Minimal Inhibitory Concentration (MIC90) of Ofloxacin and Ciprofloxacin were found 1 mg/L and 4 mg/L, respectively. There were no difference in susceptibility to Ofloxacin and Ciprofloxacin between strains which were susceptible or resistant to Standard Anti-tubercular drugs. PMID- 8421441 TI - [Evaluation of 18 epididymo-orchitis cases]. AB - Eighteen patients with epididymo-orchitis were reviewed clinically, microbiologically and serologically. While there were positive urine culture in 5 patients before treatment, only one of them had positive culture in the epididymal aspirate. The epididymo-orchitis in two patients was the complications of the brucellosis. Ofloxacin or doxycycline was used in the treatment of 16 patients for 2-3 weeks and all, except one patient were recovered completely. The cases secondary to the brucellosis were treated with rifampicin plus doxycycline combination for 6 weeks. After treatment, no positive urine cultures were noted in all patients. As a result the epididymal aspiration to clarify etiological agent is not necessarily needed and empirical treatment may be generally curative. PMID- 8421442 TI - [Chlamydia trachomatis in the eyes and sexual partners of women with genital chlamydial infections]. AB - Conjunctival scrapings from 24 women found to be positive for genital Chlamydia trachomatis previously and urethral swabs from their sexual partners were obtained and processed with direct fluorescent antibody technique. 8.3% of the conjunctival scrapings and 33.3% of the urethral specimens were found positive for C. trachomatis. None of the female patients had ocular complaints or signs of conjunctivitis and only two of the male subjects had urogenital symptoms. PMID- 8421443 TI - [Hepatitis B marker seropositivity in prostitutes using ELISA]. AB - In this study, HBsAg, Anti-HBc and Anti-HBs have been investigated in 172 prostitutes and 50 healthy women with ELISA. Anti-HBc seropositivity was increasing with age in prostitutes. No significant difference was found between Hepatitis-B marker seropositivity and working period of prostitutes. In prostitutes, 6.3% HBsAg, 62.7% Anti-HBc, 50.5% Anti-HBs seropositivity and 69.1% total seropositivity was found. Statistically, except HBsAg seropositivity, seropositivity in prostitutes was higher than the control group. Therefore, prostitutes are an important risk group for Hepatitis-B infection. PMID- 8421444 TI - [Histopathological examinations of skin biopsy specimens from patients with acute HAV and HBV infections]. AB - Punch skin biopsy is performed from 35 patients who are hospitalized with the diagnosis of acute viral hepatitis (AVH) at clinical examination and to 13 control groups. Thirteen of the 35 patients (37.4%) are diagnosed as Hepatitis A (HAV) infection, and 22 (62.6%) of them are diagnosed as Hepatitis B (HBV) infection. The histopathological examination of the skin biopsy specimens under light microscope leukocytoclastic vasculitis is found only in one case in the group of HBV infection. PMID- 8421445 TI - [Nose and throat carriage in "food handlers"]. AB - From December 1991 to February 1992, 450 personnel have been investigated for pathogen microorganisms in their nose and throat. The study was performed in the Infectious Diseases Section of Gulhane Military Medical Academy. Pathogen microorganisms have been isolated from 54 nose culture (12%) and 6 throat culture (1.33%). In one person pathogen microorganisms have been isolated from his nose and throat. The difference between the two groups (The nose and throat cultures) was significant at p = 0.001 by Fisher's exact test (t = 6.414). In the nose cultures the pathogen microorganisms were Staphylococcus aureus (85.2%), Proteus vulgaris (5.6%), Proteus mirabilis (3.7%), Klebsiella pneumoniae (1.8%), Citrobacter freundii (1.8%), Group C beta-hemolytic streptococcus (1.8%) and Pseudomonas aeruginosa (1.8%). Only one (1.85%) had two different pathogen microorganisms (Staphylococcus aureus and Proteus vulgaris) in his nasal culture. In the throat cultures the pathogen microorganisms were Staphylococcus aureus (66.7%) and group A beta-hemolytic Streptococcus (50%). Only one (16.6%) had two different pathogen microorganisms (S.aureus and group A beta-hemolytic Streptococcus) in his throat culture. The same pathogen microorganisms (S.aureus) has been isolated in an only one person's nasal and throat cultures (0.22%). We treated 60 personnel who were nasal and throat carriers according to the results of antibiograms. After treatment, two still had previous pathogen microorganism (Staphylococcus aureus). These two carriers were eradicated by repeating the treatment. PMID- 8421446 TI - [The colors produced by Fusarium Link ex Fr. 1821 (Deuteromycetes) species in various carbohydrate cultures. I. Fusarium equiseti (Corda) Sacc. 1886]. AB - In this study, Fusarium equiseti (Corda) Sacc. 1886 fungus was inoculated in cultures containing glucose, maltose, galactose, mannitol, sucrose, fructose, lactose and starch and control media cultures and were observed at intervals of five days. In most of the cultures various tones of brown were observed. Our results showed a correlation when compared with literature in general no different results were observed. Three color catalogs were used for the identification of these colors. PMID- 8421447 TI - [Laboratory infections]. AB - Infection constitutes an important professional hazard for health care workers. Since the past decades infections acquired during laboratory work have caused morbidity and even mortality among laboratory employees. In this article the importance of laboratory acquired infections and some guidelines for protection are stated. PMID- 8421448 TI - [Latest developments in the serodiagnosis of tuberculosis]. AB - In the serodiagnosis of tuberculosis, we are in a period used molecular biological techniques instead of classical methods. PCR (Polymerase Chain Reaction), ELISA (enzyme-linked immunosorbent assay), gas chromatography and mass spectrophotometry are routinely used for this propose in some countries. In this review, the importance and potential of ELISA will be discussed for developing countries. PMID- 8421449 TI - Trends and projections in selected chronic disease risk factors in Missouri, 1986 2000. PMID- 8421450 TI - Toward agricultural safety and health. PMID- 8421451 TI - Alpha-1-antitrypsin deficiency: rare cause of adult cirrhosis--a case report. AB - Alpha-1-antitrypsin is a glycoprotein which inactivates proteolytic enzymes, especially neutrophil elastase. Infants deficient in this enzyme commonly develop neonatal hepatitis. In adults, the deficiency typically results in emphysema. Only rarely will an adult manifest liver disease. We present a case of adult liver cirrhosis due to Alpha-1-antitrypsin deficiency in a 63-year-old man. Manifestations of alpha-1-antitrypsin deficiency and liver disease are discussed. PMID- 8421452 TI - Hepatitis C testing: interpretation, implications, and counseling. AB - New molecular biology techniques recently identified hepatitis C virus (HCV) as the major cause of non-A, non-B hepatitis. Serologic assays for HCV specific antibodies are a significant advance, but they require cautious interpretation due to problems with the tests' sensitivity and specificity. Patients with suspected HCV infection should be thoroughly evaluated to verify the presence of infection, to exclude other forms of chronic liver disease, and to determine the extent of liver damage prior to considering treatment. PMID- 8421453 TI - The request for Tylenol. PMID- 8421454 TI - Sexual risk behaviors of STD clinic patients before and after Earvin "Magic" Johnson's HIV-infection announcement--Maryland, 1991-1992. AB - During the human immunodeficiency virus (HIV) epidemic, media and public interest have been captured periodically by accounts of persons infected with HIV. However, the effect of these stories on HIV/acquired immunodeficiency syndrome (AIDS) awareness and sexual behaviors is largely unknown. On November 7, 1991, Earvin "Magic" Johnson announced at a press conference he was infected with HIV and would be retiring from professional basketball. This report summarizes findings on the self-reported sexually transmitted disease (STD)/HIV sexual risk behaviors of patients of an STD clinic in a Maryland suburb of Washington, D.C., during the 14 weeks before and the 14 weeks after Johnson announced he was infected with HIV. PMID- 8421455 TI - Probable transmission of multidrug-resistant tuberculosis in a correctional facility--California. AB - During 1990-1991, an inmate in a California prison spent a total of 6 months in the prison infirmary with multidrug-resistant tuberculosis (MDR-TB). As a result, from November 1990 through March 1991, 11 of the 21 prison infirmary physicians and nurses underwent tuberculin skin testing; two persons could be documented as newly positive. In addition, two correctional officers (from an unknown number tested) also had newly positive tuberculin skin tests. State and local departments of health, industrial relations, and corrections investigated the possibility of nosocomial TB transmission from inmates to staff. This report presents the findings from their investigation. PMID- 8421456 TI - Update: influenza activity--United States, 1992-93 season. AB - From September 27, 1992, through January 19, 1993, 344 influenza virus isolates were reported in 29 states (Figure 1). The number of reported isolates began to increase in early December and continued to increase during the first 2 weeks of January. PMID- 8421457 TI - Differences between anonymous and confidential registrants for HIV testing- Seattle, 1986-1992. AB - Human immunodeficiency virus (HIV) counseling and testing is a major component of the public health effort to contain the HIV/acquired immunodeficiency syndrome (AIDS) epidemic. However, persons may avoid HIV testing in part because they fear discrimination and legal sanctions if their drug use, sexual behavior, or test results became public (1,2). Anonymous testing has been offered to address these fears, but it is not clear whether anonymous testing, compared with confidential testing, actually results in testing more persons at risk for or infected with HIV. This report, using registration data from a freestanding HIV-testing clinic in Seattle, compares demographic, behavioral, and serologic characteristics of anonymous and confidential registrants during 1986-1992. PMID- 8421458 TI - Public health focus: effectiveness of rollover protective structures for preventing injuries associated with agricultural tractors. AB - Agriculture ranks fourth among U.S. industries for work-related fatalities (1). Fatalities associated with agricultural machinery commonly involve farm tractors, and rollover incidents (i.e., the tractor tips sideways or backward and overturns, crushing the operator) account for 46% (Minnesota) to 76% (Georgia) of all farm tractor-related fatalities (2). Annually, agricultural tractor rollovers result in approximately 132 work-related deaths among persons aged > or = 16 years* (3). This report summarizes information regarding the efficacy, effectiveness, and cost-effectiveness of rollover protective structures (ROPS) on agricultural tractors. PMID- 8421459 TI - Brief report: deficiency of pulmonary surfactant protein B in congenital alveolar proteinosis. PMID- 8421460 TI - Pathogenesis and treatment of gallstones. PMID- 8421461 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 6-1993. A 69-year-old woman with a sclerotic lesion of the femur and pulmonary nodules. PMID- 8421462 TI - A new therapy for the adult respiratory distress syndrome. PMID- 8421463 TI - Pandemonium in the modern hospital. PMID- 8421464 TI - Screening and informed consent. PMID- 8421465 TI - The pathogenesis and treatment of kidney stones. PMID- 8421466 TI - The pathogenesis and treatment of kidney stones. PMID- 8421468 TI - The pathogenesis and treatment of kidney stones. PMID- 8421467 TI - The pathogenesis and treatment of kidney stones. PMID- 8421469 TI - Replacement of central vascular catheters. PMID- 8421470 TI - Replacement of central vascular catheters. PMID- 8421471 TI - Replacement of central vascular catheters. PMID- 8421472 TI - High frequency of the R117H cystic fibrosis mutation in patients with congenital absence of the vas deferens. PMID- 8421473 TI - Integration of cellular and molecular biology with clinical research in cardiology. PMID- 8421474 TI - Recombinant factor VIII for the treatment of previously untreated patients with hemophilia A. Safety, efficacy, and development of inhibitors. Kogenate Previously Untreated Patient Study Group. AB - BACKGROUND: Although methods of viral attenuation in plasma-derived clotting factor concentrates have improved, there is still a possibility that such concentrates may transmit certain blood-borne viruses. For this reason, the use of recombinant DNA-derived factor VIII (which is virus-free) to treat hemophilia A has generated considerable interest. METHODS: We conducted a multicenter trial in previously untreated children with hemophilia A. They received recombinant factor VIII for all treatment or for prophylaxis and were evaluated at their respective clinics at intervals of no more than three months. RESULTS: Between January 1, 1989, and July 1, 1992, 95 patients who could be evaluated received recombinant factor VIII. By September 1, 1992, they had received the concentrate exclusively for 2.4 months to 3.5 years (median, 1.5 years). All responded well, with no treatment failures. A total of 3315 infusions were administered; there were three reports of minor adverse reactions. Inhibitor antibodies to factor VIII developed in 16 of 81 patients tested for them, after a median of nine days of exposure to factor VIII treatment. Inhibitor titers were or became low in 9 of the 16 patients despite continued episodic treatment with the concentrate. Inhibitors disappeared completely in 4 patients and remained at a low level (< 10 Bethesda units) in 5 patients receiving episodic treatment. CONCLUSIONS: Transient or low levels of inhibitor, as observed in this study, may represent part of the natural history of hemophilia in infants. In view of the transient nature and lower concentration of the inhibitors detected and the generally satisfactory response to treatment, the benefits of recombinant factor VIII for the treatment of hemophilia seem to outweigh the risks. PMID- 8421475 TI - Effect of calcium supplementation on bone loss in postmenopausal women. AB - BACKGROUND: The use of calcium supplements slows bone loss in the forearm and has a beneficial effect on the axial bone density of women in late menopause whose calcium intake is less than 400 mg per day. However, the effect of a calcium supplement of 1000 mg per day on the axial bone density of postmenopausal women with higher calcium intakes is not known. METHODS: We studied 122 normal women at least three years after they had reached menopause who had a mean dietary calcium intake of 750 mg per day. The women were randomly assigned to treatment with either calcium (1000 mg per day) or placebo for two years. The bone mineral density of the total body, lumbar spine, and proximal femur was measured every six months by dual-energy x-ray absorptiometry. Serum and urine indexes of calcium metabolism were measured at base line and after 3, 12, and 24 months. RESULTS: The mean (+/- SE) rate of loss of total-body bone mineral density was reduced by 43 percent in the calcium group (-0.0055 +/- 0.0010 g per square centimeter per year) as compared with the placebo group (-0.0097 +/- 0.0010 g per square centimeter per year, P = 0.005). The rate of loss of bone mineral density was reduced by 35 percent in the legs (P = 0.02), and loss was eliminated in the trunk (P = 0.04). Calcium use was of significant benefit in the lumbar spine (P = 0.04), and in Ward's triangle the rate of loss was reduced by 67 percent (P = 0.04). Calcium supplementation had a similar effect whether dietary calcium intake was above or below the mean value for the group. Serum parathyroid hormone concentrations tended to be lower in the calcium group, as were urinary hydroxyproline excretion and serum alkaline phosphatase concentrations. CONCLUSIONS: Calcium supplementation significantly slowed axial and appendicular bone loss in normal post-menopausal women. PMID- 8421476 TI - Direct diagnosis of myotonic dystrophy with a disease-specific DNA marker. AB - BACKGROUND: Myotonic dystrophy is the most common inherited form of muscular dystrophy affecting adults. Its symptoms are not confined to muscle, and variability in their nature and in the patient's age at their onset can make diagnosis difficult. A specific unstable DNA sequence associated with myotonic dystrophy has recently been identified. We describe the use of a DNA probe (p5B1.4) that can detect this mutation directly, improving the accuracy and speed of diagnosis. METHODS: We analyzed DNA extracted from the peripheral-blood lymphocytes of 112 unrelated patients with myotonic dystrophy and their families, using molecular genetic techniques. Southern blot analysis and amplification with the polymerase chain reaction were used to determine the extent of expansion of the unstable DNA sequence. RESULTS: Probe p5B1.4 allowed direct identification of the myotonic dystrophy mutation in 108 of the 112 unrelated patients. In three families for whom the clinical and genetic data obtained with linked probes were ambiguous, the probe identified persons at risk for symptoms of this disorder and demonstrated that a possible sporadic case of myotonic dystrophy was familial. In one of these families the size of the unstable myotonic dystrophy-specific fragment decreased on transmission to offspring, who remained asymptomatic. CONCLUSIONS: The diagnosis of myotonic dystrophy is improved by the use of a probe that detects directly the mutation responsible for this disorder. PMID- 8421477 TI - Brief report: reverse mutation in myotonic dystrophy. PMID- 8421478 TI - Gynecomastia. PMID- 8421479 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 7-1993. A six-year-old boy with multiple bone lesions, repeated fractures, and sexual precocity. PMID- 8421480 TI - Thinking straight about calcium. PMID- 8421481 TI - Immune-complex glomerulonephritis--deposits plus interest. PMID- 8421482 TI - When life support is questioned early in the care of patients with cervical-level quadriplegia. PMID- 8421483 TI - Do medical journals suppress information? PMID- 8421484 TI - Cardiac structure and function in HIV-infected children. PMID- 8421485 TI - Survival of ethylene glycol poisoning with profound acidemia. PMID- 8421486 TI - More on snake-venom and insect-venom extractors. PMID- 8421487 TI - Adventures with a vulnerable knee. PMID- 8421488 TI - Cell biology, Sticking to the point. PMID- 8421489 TI - Healy: in or out at NIH? PMID- 8421490 TI - Synthetic chemistry. Nature's anticancer agents. PMID- 8421491 TI - Ribosomal RNA trees misleading? PMID- 8421492 TI - A palatal rete in the right whale? PMID- 8421493 TI - Checkpoint check. PMID- 8421494 TI - A synaptic model of memory: long-term potentiation in the hippocampus. AB - Long-term potentiation of synaptic transmission in the hippocampus is the primary experimental model for investigating the synaptic basis of learning and memory in vertebrates. The best understood form of long-term potentiation is induced by the activation of the N-methyl-D-aspartate receptor complex. This subtype of glutamate receptor endows long-term potentiation with Hebbian characteristics, and allows electrical events at the postsynaptic membrane to be transduced into chemical signals which, in turn, are thought to activate both pre- and postsynaptic mechanisms to generate a persistent increase in synaptic strength. PMID- 8421495 TI - Fast forward for gene therapy. PMID- 8421496 TI - FGF-4 and BMP-2 have opposite effects on limb growth. AB - Limb development is dependent on epithelial-mesenchymal interactions. The apical ectodermal ridge (AER), a specialized epithelium at the limb tip, stimulates proliferation of underlying mesenchyme, causing directed limb outgrowth (for review see ref. 2). Several genes are expressed in the mouse AER, including Fgf-4 (fibroblast growth factor-4) and Bmp-2 (bone morphogenetic protein-2), both of which encode secreted signalling molecules. Using a culture system developed to explore the function of molecules produced by the AER, we have shown that FGF-4 protein stimulates proliferation of mesenchyme in the early mouse limb-bud. This suggests that FGF-4 serves that major function of the AER. In contrast, BMP-2 inhibits limb growth, suggesting that as a result the AER may serve a hitherto unrecognized inhibitory function. Furthermore, the extent of limb outgrowth can be modulated by mixing the two signalling molecules, suggesting that limb growth is regulated by a combination of stimulatory and inhibitory signals from the AER. PMID- 8421497 TI - A gene for the mouse pink-eyed dilution locus and for human type II oculocutaneous albinism. AB - The mouse pink-eyed dilution (p) locus on chromosome 7 is associated with defects of skin, eye and coat pigmentation. Mutations at p cause a reduction of eumelanin (black-brown) pigment and altered morphology of black pigment granules (eumelanosomes), but have little effect on pheomelanin (yellow-red) pigment. We show here that the human complementary DNA DN10, linked to the p locus in mice, identifies the human homologue (P) of the mouse p gene, and appears to encode an integral membrane transporter protein. The expression pattern of this gene in various p mutant mice correlates with the pigmentation phenotype; moreover, an abnormally sized messenger RNA is detected in one mutant, p(un), which reverts to the normal size in p(un) revertants. The human P gene corresponds to the D15S12 locus within the chromosome segment 15q11-q13, which is typically deleted in patients with Prader-Willi and Angelman syndrome (see ref. 5 for review). These disorders are phenotypically distinct, depending on the parent of origin of the deleted chromosome, but both syndromes are often associated with hypopigmentation of the skin, hair and eyes (see ref. 8 for review), and deletion of the P gene may be responsible for this hypopigmentation. In addition, we report a mutation in both copies of the human P gene in one case of tyrosinase-positive (type II) oculocutaneous albinism, recently linked to 15q11-q13 (ref. 9). PMID- 8421498 TI - Adhesion of epidermal Langerhans cells to keratinocytes mediated by E-cadherin. AB - Langerhans cells (LC) are the principal accessory cells present in epidermis. Because LC have limited capacity for self-renewal, epidermis is continually repopulated by as-yet uncharacterized bone marrow-derived LC progenitors. In addition, although LC persist in epidermis for extended periods, LC are induced to migrate from skin to regional lymph nodes after antigen exposure. To begin to elucidate mechanisms involved in LC trafficking, we characterized LC-keratinocyte (KC) interactions. Here we report that fresh murine LC express cadherins, and that LC adhere to KC in vitro through E-cadherin. Cultured LC (which may bear a phenotypic and functional relationship to LC that have migrated to lymph nodes) express lower levels of E-cadherin and exhibit decreased affinity for KC. These results suggest that expression of E-cadherin by LC promotes persistence of these cells in epidermis, and that cadherins may play important and unanticipated roles in interactions between leukocytes and epithelia. PMID- 8421499 TI - Metal ion catalysis in the Tetrahymena ribozyme reaction. AB - All catalytic RNAs (ribozymes) require or are stimulated by divalent metal ions, but it has been difficult to separate the contribution of these metal ions to formation of the RNA tertiary structure from a more direct role in catalysis. The Tetrahymena ribozyme catalyses cleavage of exogenous RNA or DNA substrates with an absolute requirement for Mg2+ or Mn2+ (ref. 6). A DNA substrate, in which the bridging 3' oxygen atom at the cleavage site is replaced by sulphur, is cleaved by the ribozyme about 1,000 times more slowly than the corresponding unmodified DNA substrate when Mg2+ is present as the only divalent metal ion. But addition of Mn2+ or Zn2+ to the reaction relieves this negative effect, with the 3' S-P bond being cleaved nearly as fast as the 3' O-P bond. Considering that Mn2+ and Zn2+ coordinate sulphur more strongly than Mg2+ does, these results indicate that the metal ion contributes directly to catalysis by coordination to the 3' oxygen atom in the transition state, presumably stabilizing the developing negative charge on the leaving group. We conclude that the Tetrahymena ribozyme is a metalloenzyme, with mechanistic similarities to several protein enzymes. PMID- 8421500 TI - Solution structure of the cyclosporin A/cyclophilin complex by NMR. AB - Cyclosporin A, a cyclic undecapeptide, is a potent immunosuppressant that binds to a peptidyl-prolyl cis-trans isomerase of 165 amino acids, cyclophilin. The cyclosporin A/cyclophilin complex inhibits the calcium- and calmodulin-dependent phosphatase, calcineurin, resulting in a failure to activate genes encoding interleukin-2 and other lymphokines. The three-dimensional structures of uncomplexed cyclophilin, a tetrapeptide/cyclophilin complex, and cyclosporin A when bound to cyclophilin have been reported. However, the structure of the cyclosporin A/cyclophilin complex has not been determined. Here we present the solution structure of the cyclosporin A/cyclophilin complex obtained by heteronuclear three-dimensional NMR spectroscopy. The structure, one of the largest determined by NMR, differs from proposed models of the complex and is analysed in terms of the binding interactions and structure/activity relationships for CsA analogues. PMID- 8421501 TI - X-ray structure of a decameric cyclophilin-cyclosporin crystal complex. AB - Human cyclophilin A (CypA), a ubiquitous intracellular protein of 165 amino acids, is the major receptor for the cyclic undecapeptide immunosuppressant drug cyclosporin A (CsA), which prevents allograft rejection after transplant surgery and is efficacious in the field of autoimmune diseases. CsA prevents T-cell proliferation by blocking the calcium-activated pathway leading to interleukin-2 transcription. Besides their ability to bind CsA, the cyclophilin isoforms also have peptidyl-prolyl isomerase activity and enhance the rate of protein folding. The macrolide FK506 acts similarly to CsA and its cognate receptor FKBP also has peptidyl-prolyl isomerase activity. Inhibition of this enzymatic activity alone is not sufficient to achieve immunosuppression. A direct molecular interaction between the drug-immunophilin complex (CsA-CypA, or FK506-FKBP) and the phosphatase calcineurin, is responsible for modulating the T-cell receptor signal transduction pathway. Here we describe the crystal structure of a decameric CypA CsA complex. The crystallographic asymmetric unit is composed of a pentamer of 1:1 cyclophilin-cyclosporin complexes of rather exact non-crystallographic fivefold symmetry. The 2.8 A electron density map is of high quality. The five independent cyclosporin molecules are clearly identifiable, providing an unambiguous picture of the detailed interactions between a peptide drug and its receptor. It broadly confirms the results of previous NMR, X-ray and modelling studies, but provides further important structural details which will be of use in the design of drugs that are analogues of CsA. PMID- 8421502 TI - Active-centre torsion-angle strain revealed in 1.6 A-resolution structure of histidine-containing phosphocarrier protein. AB - The histidine-containing phosphocarrier protein (HPr) is a central component of the phosphoenolpyruvate: sugar phosphotransferase system that transports carbohydrates across the cell membrane of bacteria. A typical phosphotransfer sequence is phosphoenolpyruvate-->enzyme I-->HPr-->enzyme II/IIIsugar-->sugar. This is thermodynamically favourable owing to the participation of the high energy phosphoenolpyruvate. We report here the structure of HPr from Streptococcus faecalis determined at 1.6 A resolution. Remarkable disallowed Ramachandran torsion angles at the active centre, revealed by the X-ray structure, demonstrate a unique example of torsion-angle strain that is probably directly involved in protein function. During phosphorylation, the active-centre torsion-angle strain should facilitate the phosphotransfer reaction by lowering the activation-energy barrier. A recently reported Bacillus subtilis HPr structure, which represents the phosphorylated state of HPr with no torsion-angle strain, provides direct evidence supporting our hypothesis that torsion-angle strain plays a direct part in the function of HPr. An HPr phosphotransfer cycling mechanism is proposed, based primarily on the structures of HPr and other phosphotransferase system proteins. PMID- 8421503 TI - Cloning the Menkes disease gene. AB - Three groups have successfully isolated the gene responsible for the X-linked Menkes disease, heralding great promise for our understanding of copper metabolism and for diagnosis of the disorder. PMID- 8421504 TI - Cuenod will head Human Frontier programme. PMID- 8421505 TI - French study sparks debate on informed consent laws. PMID- 8421506 TI - Medical council of Canada broadens role. PMID- 8421507 TI - Protesters target European animal patents. PMID- 8421508 TI - Consciousness in other words. PMID- 8421509 TI - Creative reviews. PMID- 8421510 TI - Future of Baer Museum. PMID- 8421511 TI - Unreconstructed man. PMID- 8421512 TI - Unreconstructed man. PMID- 8421513 TI - Backwardness of human neuroanatomy. AB - To interpret the activity of living human brains, their neuroanatomy must be known in detail. New techniques to do this are urgently needed, since most of the methods now used on monkeys cannot be used on humans. PMID- 8421514 TI - Visual system. Vision in blind mole rats. PMID- 8421515 TI - Cell motility. Variations on the theme of movement. PMID- 8421516 TI - Developmental biology. Transforming frogs and flies. PMID- 8421517 TI - A word in your protein. PMID- 8421518 TI - Neurotoxicity of beta-amyloid. PMID- 8421519 TI - Photoinduced electron transport across a lipid bilayer mediated by C70. AB - Electron transport across a membrane is central to photosynthesis, to mitochondrial respiration and to the design of molecular systems for solar energy conversion. Relatively few synthetic molecules, however, have been shown to facilitate transport of electrons across a lipid bilayer. We report here that C70 can act as both a photosensitizer for electron transfer from a donor molecule and a mediator for electron transport across a lipid bilayer membrane. The steady state photocurrent density obtained from the C70-bilayer system is about 40 times higher, at comparable light intensities, than that of the carotene porphyrinquinone system, previously the most efficient artificial system. The C70 bilayer system has a quantum yield of about 0.04, while the stability (tens of minutes) and turnover number (electrons transported per C70 before decay) of 10(3) are one to three orders of magnitude greater than those of other systems. We anticipate that other higher fullerenes may also provide the basis for efficient transmembrane electron-transport systems. PMID- 8421520 TI - Disease resistance results from foreign phytoalexin expression in a novel plant. AB - Although phytoalexins have long been inferred to be important in the defence of plants against fungal infection, there are few reports showing that they provide resistance to infection. Several plants, including grapevine, synthesize the stilbene-type phytoalexin resveratrol when attacked by pathogens. Stilbenes with fungicidal potential are formed in several unrelated plant species, such as peanut (Arachis hypogaea), grapevine (Vitis vinifera) and pine (Pinus sylvestris). Stilbene biosynthesis only specifically requires the presence of stilbene synthase. Furthermore, the precursor molecules for the formation of hydroxy-stilbenes are malonyl-CoA and p-coumaroyl-CoA, both present in plants. To investigate the potential of stilbene biosynthetic genes in a strategy of engineering pathogen resistance, we isolated stilbene synthase genes from grapevine, where they are expressed at a high level, and transferred them into tobacco. We report here that regenerated tobacco plants containing these genes are more resistant to infection by Botrytis cinerea. This is, to our knowledge, the first report of increased disease resistance in transgenic plants based on an additional foreign phytoalexin. PMID- 8421521 TI - Novel GABA responses from rod-driven retinal horizontal cells. AB - gamma-Aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the central nervous system. Two classes of GABA receptors (GABAA and GABAB) have been identified. GABAA receptors are ligand-gated chloride channels that are competitively antagonized by bicuculline, noncompetitively blocked by picrotoxin, and often allosterically modulated by barbiturates and benzodiazepines. GABAB receptors regulate potassium and calcium channels through G-protein and intracellular second-messenger pathways, are selectively activated by baclofen, and are antagonized by phaclofen and 2-hydroxysaclofen. For some years, evidence has accumulated that there are GABA receptors, especially prominent along visual pathways, which are neither antagonized by bicuculline nor activated by baclofen, but are activated by certain conformationally restricted analogues of GABA, including cis-4-aminocrotonic acid (CACA). These receptors have been designated GABAC receptors. As yet, membrane current responses from isolated neurons that reflect this novel pharmacology have not been reported, although such responses have been recorded from oocytes injected with retinal messenger RNA. Here we describe a chloride-mediated current response from isolated rod-driven horizontal cells (H4) of the white perch retina that has this novel pharmacology. PMID- 8421522 TI - Chemomechanical cycle of kinesin differs from that of myosin. AB - Motor proteins move unidirectionally along cytoskeletal polymers by coupling translocation to cycles of ATP hydrolysis. The energy from ATP is required both to generate force and to dissociate the motor-filament complex in order to begin a new chemomechanical cycle. For myosin, force production is associated with phosphate release following ATP hydrolysis, whereas dissociation of actomyosin is tightly coupled to the binding of ATP. Dynein, a microtubule motor, uses a similar cycle, suggesting that all cytoskeletal motors might operate by a common mechanism. Here we investigate kinesin's chemomechanical cycle by assaying microtubule movement by single kinesin molecules when intermediate states in the hydrolysis cycle are prolonged with ATP analogues or inhibitors. In contrast to myosin and dynein, kinesin with bound ADP dissociates from microtubules during translocation, whereas kinesin with unhydrolysed nucleotide remains tightly associated with the polymer. These findings imply that kinesin converts ATP energy into mechanical work by a pathway distinct from that of myosin or dynein. PMID- 8421523 TI - Arsenical-resistant trypanosomes lack an unusual adenosine transporter. AB - The melaminophenyl arsenical melarsoprol is still used to treat African sleeping sickness, a disease caused by parasitic protozoa of the Trypanosoma brucei subgroup. Based on the observation that melamine antagonizes the trypanocidal activity of this class of drugs, we investigated whether other physiological compounds could compete for the same receptor. Here we report that the in vitro trypanolytic effect of melarsen oxide can be specifically abrogated by adenine, adenosine and dipyridamole, all of which compete for uptake by an adenosine transporter. Melarsen-sensitive trypanosomes have two high-affinity adenosine transport systems: a P1 type, which also transports inosine; and a P2 type, which also transports adenine and the melaminophenyl arsenicals. Melarsen-resistant trypanosomes lack P2 adenosine transport, suggesting that resistance to these arsenicals is due to loss of uptake. PMID- 8421524 TI - Ordered duplex RNA controls capsid architecture in an icosahedral animal virus. AB - Small spherical viruses are among the simplest replicating systems in biology, yet the factors affecting their assembly, stability and disassembly are still poorly understood. A molecular switch is required for the assembly of icosahedral virus particles containing more than 60 identical subunits because strict symmetry cannot be maintained in subunit packing. All previously reported viruses with this type of structure use a portion of the capsid protein to regulate interactions between chemically equivalent but structurally distinct interfaces. We have investigated the T = 3 quasiequivalent nodaviruses, which are small non enveloped viruses with a single-stranded RNA genome that infect insects, mice and fish. They undergo a well-characterized series of steps in assembly and maturation, which in some respects are similar to the picornaviruses, despite their different capsid architecture. Here we report the X-ray structure of Flock House virus at 3.0 A resolution, which reveals an ordered RNA duplex of 20 nucleotides and a protein segment that control the subunit interactions in this animal virus. The RNA interacts with a helical protein domain of the subunit that lies inside the capsid shell. One of the helices that binds the RNA is part of a 44-amino-acid polypeptide which is autocatalytically cleaved from the initial subunit translation product after virion assembly. The structure indicates that RNA associated with the cleaved polypeptide may be important in the infection process. PMID- 8421525 TI - Double-helical RNA in satellite tobacco mosaic virus. AB - Satellite tobacco mosaic virus (STMV) is the spherical satellite to an obligatory rod-shaped helper tobacco mosaic virus (TMV), which is required for replication. STMV has 60 protein subunits of M(r) 17,500 on a T = 1 icosahedral capsid containing a single-stranded RNA genome of 1,059 bases. STMV appears similar to another virus, STNV, but is approximately 20 per cent smaller. It shows no amino acid homology or immunological cross-reactivity with either STNV or its host TMV. Here we report the X-ray crystal structure of STMV, which shows that the coat protein of STMV contains a 'Swiss roll' beta-barrel. An amino-terminal strand extends more than 60A and is primarily responsible for quaternary interactions. Each capsid dimer is associated with a segment of genomic RNA double helix comprising seven base pairs. The dyad of each protein dimer is coincident with that of the central base pair of the associated RNA segment whose helix axis is directed along an icosahedral edge. Protein-nucleic acid interactions are extensive. The RNA helices, which have additional stacked bases at their 3' termini, differ significantly from canonical nucleic acid helical forms. PMID- 8421526 TI - Evolution in vitro of an RNA enzyme with altered metal dependence. AB - The Tetrahymena group I ribozyme catalyses a sequence-specific phosphodiester cleavage reaction on an external RNA oligonucleotide substrate in the presence of a divalent metal cation cofactor. This reaction proceeds readily with either Mg2+ or Mn2+, but no detectable reaction has been reported when other divalent cations are used as the sole cofactor. Cations such as Ca2+, Sr2+ and Ba2+ can stabilize the correct folded conformation of the ribozyme, thereby partially alleviating the Mg2+ or Mn2+ requirement. But catalysis by the ribozyme involves coordination of either Mg2+ or Mn2+ at the active site, resulting in an overall requirement for one of these two cations. Here we use an in vitro evolution process to obtain variants of the Tetrahymena ribozyme that are capable of cleaving an RNA substrate in reaction mixtures containing Ca2+ as the divalent cation. These findings extend the range of different chemical environments available to RNA enzymes and illustrate the power of in vitro evolution in generating macromolecular catalysts with desired properties. PMID- 8421527 TI - [A little less sodium is not without hazard]. PMID- 8421528 TI - [DNA and dystrophin analysis in Duchenne's and Becker's disease; benefit to the patient]. PMID- 8421529 TI - [Controversies in preventive health care. II. Preconceptional and prenatal care]. PMID- 8421530 TI - [Application of combined DNA and dystrophin protein analysis in the diagnosis of Duchenne's and Becker's muscular dystrophy in 102 Dutch patients]. AB - The recent progress in molecular genetic studies on Duchenne and Becker muscular dystrophy (DMD, BMD) had an important spin-off for our diagnostic abilities of both muscle disease. The mapping and isolation of the DMD gene which codes for the 427 kD cytoskeletal protein dystrophin made it possible to diagnose 80-85% of the patients by means of DNA analysis. At present, most of the remaining 15-20% of the patients can be diagnosed by protein analysis. In this report we describe the analysis of dystrophin in a group of 102 Dutch patients with muscular dystrophies. An immunohistochemical and immunobiochemical study of dystrophin was performed on muscle tissue, partly integrated with DNA analysis. In this study we underline the value of dystrophin analysis in all patients suspected of DMD, BMD or other muscular dystrophies, particularly in those without detectable DNA mutations. By means of integrated DNA/dystrophin analysis 98% of the DMD patients and 90% of th BMD patients and their families can now be provided with an unambiguous diagnosis. In particular, discrimination between BMD and other muscular dystrophies has strongly improved. PMID- 8421531 TI - [The value of cytological studies in nodular goiter: results in 495 patients]. AB - The accuracy of clinical diagnosis and fine-needle aspiration biopsy (FNAB) was evaluated in a total of 495 patients of whom 183 were operated upon within 6 months after FNAB and 312 were not. Operated patients were divided into three subgroups with high, moderate or low suspicion of malignant neoplasms on clinical grounds. Histological examination revealed an overall malignant neoplasm rate of 23%. The rate, i.e. the positive predictive value, was 74% for the subgroup with high clinical suspicion, vs 14% and 10% respectively for the subgroups with moderate and low clinical suspicion. The sensitivity of a high clinical suspicion was 60% and the specificity 93%. The overall sensitivity of FNAB was 93% in the operated group and probably not less than 87% in all patients studied; specificity was 71% and 84% respectively for these groups. The overall positive predictive value of a positive cytology result (malignant or uncertain) was 48%. In the subgroups with moderate or low clinical suspicion which are more representative of a non-university setting, the average predictive value was 27%. In our opinion all patients in the group with high clinical suspicion need surgical treatment, regardless of the FNAB result; those with lower degrees of clinical suspicion and malignant or uncertain FNAB result should also undergo diagnostic surgical exploration. PMID- 8421532 TI - [The dynamic hip screw in medial fractures of the femoral neck: results in 51 patients]. AB - Femoral neck fractures are common in the elderly, especially in female patients. Pseudarthrosis and femoral head necrosis were complications with a high incidence in earlier days, when the femoral neck fracture was treated by obsolete osteosynthesis techniques. Replacement of the femoral head by an endoprosthesis was introduced in the seventies because of these complications. This hemiarthroplasty also causes many postoperative complications. With the development of the dynamic hip screw for trochanteric and subtrochanteric fractures a new technique for stabilising femoral neck fractures has been introduced. In a retrospective study (average follow-up: 3.4 years; 3 months-6 years; median: 3.6 years) of 51 patients with a femoral neck fracture who were treated with a DHS the short- and long-term postoperative complications (femoral head necrosis and pseudarthrosis) were determined. The most frequent short-term complications were bleeding (10 patients), the need for blood transfusion (10), decubitus (6), wound infection (5), urinary tract infection (5). Femoral head necrosis was seen in two patients more than 30 days after the operation. Pseudarthrosis was not observed. Because of the low complication rate, we can conclude that DHS osteosynthesis, on a correct indication, can be used as first choice in femoral neck fractures. PMID- 8421534 TI - [Albania--the Cinderella of Europe]. PMID- 8421533 TI - [3 adolescents with hip pain caused by idiopathic chondrolysis]. AB - The cases are described of three adolescents in whom hip pain was found to be due to chondrolysis. Arthroscopy revealed chronic synovitis in all three cases. It is stated that the synovitis is probably the cause of the articular damage and that the disorder should be regarded as juvenile chronic arthritis of the oligoarticular type. Treatment with intra-articular triamcinolone acetonide and in two patients with an anti-inflammatory agent as well resulted in almost complete recovery of hip function. PMID- 8421535 TI - [Albania, a country in transition]. PMID- 8421536 TI - [Abdominal aortic aneurysm as incidental finding in echographic study of the abdomen]. PMID- 8421537 TI - A systematic approach to spinal reconstruction after anterior decompression for neoplastic disease of the thoracic and lumbar spine. AB - The anterior approach to the thoracic and lumbar spine for neoplastic disease is now a well-accepted procedure, with results, for the most part, superior to those achieved with laminectomy. However, the specific indications for anterior decompression and the selection of reconstruction techniques based on the location and extent of bony destruction have received surprisingly little attention. The authors report their experience with the operative management of 33 patients with benign and malignant tumors of the thoracic and lumbar spine, using the anterior transthoracic or retroperitoneal approach. The role of stabilization and the relative indications for anterior or posterior instrumentation are emphasized. The mean age of patients was 58 years. Twenty three patients were male. Five patients had benign tumors, and the remainder had a variety of metastatic lesions. Twenty-nine patients had lower extremity motor deficits, although 25 were ambulatory preoperatively. Thirty-seven noncontiguous resections were performed in 33 patients. In 13 patients, the resected vertebral body was replaced with acrylic or bone without instrumentation; in 18, the acrylic was supplemented with anterior instrumentation; and in 6, both anterior and posterior instrumentation were used. Above T11, vertebral reconstruction techniques were used to restore stability after decompression. Between T11 and L4, anterior instrumentation was used to supplement vertebral reconstruction in all patients. Supplemental posterior instrumentation was used for three-column involvement. Motor function was stabilized or improved in 94% of patients, and 88% of patients were ambulatory postoperatively. Of 28 patients with malignant disease, 23 died after a mean survival of 10.2 months (range, 2-51 mo) and 5 are alive a mean of 34.4 months since their operation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421538 TI - The histopathological effects of the CO2 versus the KTP laser on the brain and spinal cord: a canine model. AB - Because no data are available concerning the histopathological effects of the potassium titanyl phosphate (KTP) laser on central nervous tissue, a study was performed using a canine model to compare the histopathological effects of a commonly used laser (CO2) and the KTP laser on brain and spinal cord tissue. Exposed brain and spinal cord tissue were irradiated with 0.1-s pulses (x10), with spot sizes of 1 mm (in focus) over a range of 1 to 10 W. Wedge-shaped lesions were produced with the CO2 laser, while more blunt, semilunar-shaped lesions were produced by the KTP laser. The depth and width of the lesions were proportional to the energy applied. The lesions ranged in surface diameter from 0.6 to 1.3 mm for CO2 and 0.8 to 1.6 mm for KTP lasers, respectively. The depth of the lesions varied from 0.4 to 2.0 mm for CO2 and 0.3 to 1.1 mm for KTP lesions. Histopathologically, a central zone of tissue destruction and vaporization was surrounded by a zone of coagulative necrosis, in turn surrounded peripherally by a zone of pallor. CO2-induced lesions were histologically more hemorrhagic than KTP-induced lesions. In view of the histopathological findings, the KTP laser appears as safe as the CO2 laser in terms of tissue lateral thermal change (penetration) and tissue absorption. The additional hemostatic advantage observed clinically for the KTP laser is demonstrated histologically as well. Although the wavelength of the KTP and argon laser light are similar, the histopathological effects seem to be less pigment dependent. The KTP laser seems well suited for neurosurgery and has the versatility provided by a fiberoptic delivery system. PMID- 8421539 TI - The "subdural" space: a new look at an outdated concept. AB - This review considers the structure of the meninges, as seen at the electron microscopic level, with particular emphasis on the dura-arachnoid junction and whether a naturally occurring space is found at this interface. The classic view has been that a so-called subdural space is located between the arachnoid and dura and that subdural hematomas or hygromas are the result of blood or cerebrospinal fluid accumulating in this (preexisting) space. The dura is composed of elongated, flattened fibroblasts and copious amounts of extracellular collagen. A specialized layer of fibroblasts, the dural border cell layer, is found at the dura-arachnoid junction and is characterized by flattened fibroblasts, no extracellular collagen, extracellular spaces, and few cell junctions. These features combine to create a layer of the inner dura that is structurally weak when compared with external portions of the dura and the internally located arachnoid. The arachnoid layer is composed of larger cells with numerous cell junctions, no extracellular space, and no extracellular collagen. The occurrence of many tight junctions in this layer also serves as a barrier to the movement of fluids and ions. Fibroblasts specialized to form the arachnoid trabeculae attach to the inner surface of the arachnoid layer, bridge the subarachnoid space, and surround vessels in the subarachnoid space as well as attach to pia on the surface of the brain. Under normal conditions, there is no evidence of a naturally occurring space being extant at the dura-arachnoid junction. A space may appear at this point subsequent to pathological/traumatic processes that result in tissue damage with a cleaving opening of the structurally weakest plane in the meninges--through the dural border cell layer. Furthermore, when a space does appear, it is not "subdural" in location but rather within a morphologically distinct cell layer. PMID- 8421540 TI - Recovery of vision after transcranial decompression of pituitary apoplexy characterized by third ventricular hemorrhage. AB - The authors present a case of pituitary apoplexy characterized by massive third ventricular hemorrhage. This rare complication, which caused total blindness in the patient, a 67-year-old man, was treated with transcranial decompression of the pituitary tumor and removal of a blood clot from the third ventricle. After surgery, the patient totally recovered his vision. To the best of our knowledge, this is the first case of a patient with pituitary apoplexy that ruptured into the third ventricle (diagnosed by imaging studies) who was treated by emergency transcranial decompression. PMID- 8421541 TI - Posterior dural arteriovenous malformation and medulloblastoma in an infant: case report. AB - A newborn is described who presented with heart failure from a posterior dural arteriovenous malformation and had a coexisting congenital medulloblastoma. There have been sporadic reports of arteriovenous malformation and brain neoplasms in older children and adults, and these have generally been glial tumors. This is the first known case of a combined congenital primitive neuroectodermal tumor and arteriovenous malformation in an infant. PMID- 8421542 TI - Primary carcinoma of the choroid plexus in Li-Fraumeni syndrome: case report. AB - An 8-month-old female infant with a primary carcinoma of the choroid plexus developed a rhabdomyosarcoma in the anterior chest wall at the age of 1 year and 2 months. Her mother had developed a liposarcoma in her left thigh at the age of 17 years. One of the patient's siblings had a rhabdomyosarcoma of the epipharynx at the age of 1 year. This is the fourth reported case of a choroid plexus carcinoma occurring in Li-Fraumeni syndrome. PMID- 8421543 TI - Resolution of an actinomycotic abscess with nonsurgical treatment: case report. AB - A case of actinomycotic brain abscess is presented. Conservative treatment by prolonged administration of antibiotics after needle biopsy showed complete resolution of the abscess. Previously reported cases suggest that definitive treatment requires excision or open surgical drainage of the abscess. The case presented suggests an alternative approach to treating this unusual cause of brain abscess. PMID- 8421544 TI - Functional obstruction of an antisiphon device by raised tissue capsule pressure. AB - The proper function of an antisiphon device (ASD) requires that it be exposed externally to atmospheric pressure and that its mobile membrane be free to move. A 4-year-old boy who had an ASD placed for postshunt subdural hematomas presented with functional obstruction of the ASD-symptomatic ventriculomegaly despite evidence of patency of the shunt system by radionuclide flow study. At surgery, the distal shunt system including the ASD was infused with normal saline before and after surgical exposure of the device. The pressures required to maintain a flow rate of 5 ml/h and 50 ml/h were 27 mm Hg and 30 mm Hg, respectively. After surgical exposure, these pressures fell to 0 mm Hg and 5 mm Hg, respectively, for the same infusion rates. Surgical exposure removes the effects of the tissue capsule, including the overlying skin and a collagenous sheath, restoring the external pressure to atmospheric pressure. Functional obstruction of ASDs occurs because of raised ambient pressure from the tissue capsule acting to depress the mobile membrane of the ASD, increasing its resistance to flow. This case confirms previously reported effects of subcutaneous implantation of ASDs in experimental animals and is the probable explanation for reported functional obstruction of ASDs in other patients. PMID- 8421545 TI - Spontaneous movement of bullets in the brain. AB - We report two patients in whom bullets in the brain migrated into the adjacent lateral ventricle and moved freely as a consequence of gravity. A review of the literature suggests that the spontaneous migration of intracerebral bullets is influenced by cerebral softening, the specific gravity of the bullet compared with brain tissue, and the sink function of the cerebral ventricles. In patients undergoing the surgical removal of intracerebral or intraventricular bullets, it is recommended that an x-ray be obtained after the final positioning of the head. PMID- 8421546 TI - Branches of the persistent primitive trigeminal artery--an autopsy case. AB - An adult autopsy case of the persistent primitive trigeminal artery (PTA) is reported. At the origin of the PTA in the cavernous portion of the internal carotid artery, the PTA branched into the meningohypophyseal trunk and the artery of the inferior cavernous sinus. This might indicate that these arteries were formed at the same embryonic stage. At the cisternal portion of the PTA in the posterior fossa, it also branched into the trigeminal nerve root and pons on the way to the basilar artery. The PTA might have functions for the normal structures even in the case of adults. The PTA passed through the dural foramen, which was located medial to Meckel's cave, and connected with the basilar artery. The diameter of the PTA decreased markedly at the foramen. The dural foramen might be the site of regression of the PTA during the embryonic stage. PMID- 8421547 TI - Traumatic subdural hygroma: pathology and meningeal enhancement on magnetic resonance imaging. PMID- 8421548 TI - The use of acetazolamide-enhanced regional cerebral blood flow measurement to predict risk to arteriovenous malformation patients. PMID- 8421549 TI - Elevated cortical venous pressure in hydrocephalus. PMID- 8421550 TI - Carotid endarterectomy complicated by vein patch rupture. PMID- 8421551 TI - Multiple cerebral hydatid disease: case report with magnetic resonance imaging study. PMID- 8421552 TI - Long-term observations of intracranial pressure after severe head injury. The phenomenon of secondary rise of intracranial pressure. AB - The long-term course of intracranial pressure (ICP) was studied in 53 patients from a group of 90 patients with severe head injury treated over a 3-year period. In 49 of these, ICP was significantly elevated during the observation period. The maximum in ICP was usually observed 24 to 96 hours posttrauma. A subgroup of patients developed a second rise of ICP. Such a course was observed in 15 (31%) of the 49 patients with intracranial hypertension. In these cases, ICP increased initially to 20 to 30 mm Hg but could be controlled. Thereafter, ICP was decreased again for at least 12 hours. The secondary ICP rise occurred 3 to 10 days after trauma. In six patients, intracranial hypertension became uncontrollable and eventually caused brain death. The outcome of patients with a secondary rise of ICP was worse when compared with that of patients without this complication. A cause of the secondary ICP rise could only be identified in some cases. Delayed traumatic intracerebral hemorrhage, traumatic vasospasm, hypoxia, and hyponatremia were diagnosed in seven cases. In seven other patients, the secondary ICP rise coincided with a pronounced leukocytosis, which was not associated with apparent infections. Because the occurrence and degree of a secondary rise of ICP after severe head injury are important factors affecting outcome, monitoring of ICP after severe head injury should be prolonged. PMID- 8421553 TI - Delayed and progressive brain injury in closed-head trauma: radiological demonstration. AB - The importance of delayed or secondary brain insults in the eventual outcome of closed-head trauma has been documented in experimental models. To understand this phenomenon in the clinical setting, we studied a series of head-injured patients in whom multiple cranial computed tomographic (CT) scans were obtained. Patients whose follow-up CT studies revealed new intracranial lesions or worsening, compared with admission findings, were considered to have delayed cerebral injury. One hundred forty-nine (44.5%) of 337 consecutively studied patients developed delayed brain injury. There were highly significant associations (P < 0.001) between the appearance of delayed cerebral insults and the severity of the initial brain injury, the need for cardiopulmonary resuscitation in the field, the presence of coagulopathy at admission, and subdural hematoma on the initial CT scan. In addition, delayed injury was associated (P < 0.001) with higher mortality, slowed recovery, and poorer outcome at 6 months. Delayed brain injury was not significantly associated with patient age, sex, injury mechanism, associated injury, the need for endotracheal intubation in the field, early talking, CT abnormality other than intracranial hematoma, or type of residual neurological deficits. We used multiple regression analysis to explore the relationship between severity of injury, delayed insults, and outcome. As expected, the severity of the initial brain trauma contributed significantly to neurological outcome. The presence of delayed cerebral injury makes the outcome dramatically worse for each category of initial injury severity. The relationship between initial and secondary brain injury is discussed. PMID- 8421554 TI - Surgical management of high jugular bulb in acoustic neurinoma via retrosigmoid approach. AB - Of 200 patients with acoustic neurinoma undergoing an operation via the retrosigmoid transmeatal approach in the semisitting position, 18 patients had a high jugular bulb on the tumor side. The frequency was 9%. From a neurosurgical point of view, a jugular fossa above the low border of the internal auditory canal (IAC) is classified as a high one. All 200 patients were evaluated by computed tomography with bone window reconstruction of high-resolution thin axial slices (1.5 mm). High jugular bulbs were classified into three grades as follows: Grade I, jugular bulb situated less than 1.5 mm above the low border of IAC; Grade II, jugular bulb between 1.5 and 3.0 mm above the low border of the IAC; Grade III, jugular bulb > 3 mm above the low border of IAC. There were eight patients with Grade I, six patients with Grade II, and four patients with Grade III. In these patients, in order to open the IAC without concomitant injury of the jugular bulb, the superior posterior portion of the porus was drilled away. Opening the jugular fossa was unavoidable in Grade III cases. No difference was noted in functional preservation of facial or cochlear nerve between HJB cases and normal jugular bulb cases, but HJB cases had a higher frequency of air embolism during tumor removal than did normal cases (16 versus 5%), especially Grade III cases (two of four). There was no mortality or morbidity in the cases of air embolism. Details of the surgical procedure in such cases are discussed. PMID- 8421555 TI - Correlation of microanatomical localization with postoperative survival in posterior fossa ependymomas. AB - Twenty-two surgically treated infratentorial ependymomas were analyzed according to their anatomical origins and characteristics of extension in conjunction with the microsurgical anatomy of the fourth ventricle. The correlation between tumor origin and postoperative survival of the patients was also assessed. The tumors were classified into three types according to their origins and extensions: 1) midfloor-type: tumors originating from the caudal half of the fourth ventricular floor beneath the striae medullares. After occupying the fourth ventricular cavity, they extended downward through the foramen Magendie to the upper cervical level. 2) Lateral type: tumors arising from the vestibular area and/or the lateral recess. They grew not only inferiorly but also laterally to the cerebellomedullary cistern through the cerebellomedullary fissure and the foramen of Luschka. 3) Roof type: tumors originating from the roof of the ventricle. The overall cumulative survival rates at 2, 5, and 10 years were 84, 62, and 47%, respectively. Interestingly, the lateral-type tumors showed a significantly lower 5-year cumulative survival rate and mean survival time (21% and 40 months) when compared with midfloor-type tumors (73% and 170 months). Because the tumor originates near the vital neural structures and because each type has characteristics of extension, a clear knowledge of the microanatomical relationship between the tumor and the surrounding structures would be of great benefit for improving the operative outcome of posterior fossa ependymomas. PMID- 8421556 TI - Monitoring of infectious intracranial aneurysms by sequential computed tomographic/magnetic resonance imaging studies. AB - To monitor the course of infectious intracranial aneurysms, repeated cerebral angiography has been recommended every 2 weeks during intravenous antibiotic therapy until the aneurysm has resolved or an operation has been performed. However, serial cerebral angiograms are not without some risk to the patient. We have prospectively studied five patients harboring a total of six infectious intracranial aneurysms by sequential computed tomography (CT) and/or magnetic resonance imaging (MRI) studies. All infectious aneurysms were initially identified by cerebral angiography and were treated with 6 to 8 weeks of intravenous antibiotics. The aneurysm size ranged from 4 to 10 mm. Sequential CT scans and/or MRI studies were obtained at 2- to 3-week intervals to monitor the course of the aneurysms. Three aneurysms enlarged during antibiotic therapy, and one remained unchanged in size. These four aneurysms were treated surgically. The two remaining aneurysms resolved with intravenous antibiotic therapy. Cerebral angiograms were obtained routinely preoperatively and were used to verify the resolution of the infectious aneurysms when they were no longer visible on CT or MRI. On the basis of this prospective study, we conclude that sequential thin slice CT and/or MRI can effectively and safely monitor the course of infectious intracranial aneurysms once identified by cerebral angiography. This may reduce the need for serial angiography and reduce the ultimate risk in the management of infectious intracranial aneurysms. PMID- 8421557 TI - Sonic stereometry in microsurgical procedures for deep-seated brain tumors and vascular malformations. AB - A frameless computerized navigating system was successfully employed in 20 cases of open microsurgical operations on deep-seated brain tumors and vascular malformations. Localization in space was made by measuring the traveling time of sonic waves (24 kHz) in air. This allowed the construction of mechanically simple, lightweight freehand targeting instruments. The tips can be localized with a measuring accuracy of +/- 1 mm intracranially on the basis of computed tomographic scans. This thoroughly redesigned sonic stereometrical device is being optimized for everyday use; magnetic resonance imaging, digital substraction angiography, and other diagnostic modalities will be implemented. Some perspectives on "computer-aided (neuro)surgery" are discussed. PMID- 8421558 TI - The prevention of oxyhemoglobin-induced endothelial and smooth muscle cytoskeletal injury by deferoxamine. AB - The oxidized breakdown products of hemoglobin are important in the pathogenesis of cerebral vasospasm because of their effects on the endothelium and the smooth muscle of the arterial wall. Cytoskeletal changes in cultured vascular cells are sensitive indicators of oxidative injury. Cultured endothelial cells and smooth muscle cells showed a dose-related disruption of the cytoskeleton, particularly the F-actin and vimentin filaments, when exposed to 10(-5) M oxyhemoglobin. The cytoskeletal injury was prevented by the addition of 10(-3) M deferoxamine or 1% albumin. These experiments support a role for deferoxamine in the pharmacological treatment of vasospasm. Furthermore, cytoskeletal studies of cultured arterial endothelial and smooth muscle cells provide a novel in vitro approach by which to study the cellular mechanisms of oxidant injury initiated by the breakdown products of hemoglobin. PMID- 8421559 TI - Regeneration of the rat carotid artery after clipping injury. Part I. A morphological study. AB - We investigated the natural course of the morphological regeneration of the endothelium and smooth muscle of the rat carotid artery after clipping injury. Vascular damage was produced by clipping the right carotid arteries of Wistar rats. Endothelial regeneration was confirmed by the injection of Evans blue dye and the detection of factor VIII-related antigen. The volume of the smooth muscle cell layer and the luminal size were measured by computer-assisted morphometric analysis. Immediately after arterial injury, Evans blue dye freely permeated the smooth muscle layer, suggesting that complete endothelial denudation had occurred. Endothelial regrowth started within 24 hours and was fastest on the third and fourth days after injury. The endothelial injury was repaired within 5 days. The area of the smooth muscle layer did not change immediately after clipping injury, but it gradually increased within a month. The luminal area of the injured artery increased during the 3-month recovery period. These findings suggest that endothelial regrowth is completed within a week after clipping injury, whereas smooth muscle cell regrowth is slower. In addition, arteriosclerotic luminal narrowing did not occur during recovery of the rat carotid artery from clipping injury. PMID- 8421560 TI - Regeneration of the rat carotid artery after clipping injury. Part II. A pharmacological study. AB - This study investigated the natural course of functional recovery of the vascular endothelium and smooth muscle after clipping injury of the rat carotid artery. Vascular injury was induced by clipping the right carotid arteries of Wistar rats. The contractile response to KCl, serotonin (5-hydroxytryptamine), and norepinephrine was decreased immediately after arterial injury. The response to KCl and serotonin recovered within 8 weeks, whereas the response to norepinephrine recovered after 12 weeks. Endothelium-dependent relaxation also disappeared immediately after clipping injury, but the recovery of relaxation in response to acetylcholine and adenosine triphosphate was observed within 1 week. Four weeks after clipping injury, higher doses of acetylcholine induced slight arterial contraction. These findings suggest that the recovery of smooth muscle contractility was slower than the process of endothelial regeneration in the rat carotid artery after clipping injury. Endothelium-dependent relaxation recovered within only a week, although the characteristics of the arterial cholinergic receptors may have changed in the chronic recovery stage. PMID- 8421561 TI - Clinical predictors of abnormality disclosed by computed tomography after mild head trauma. AB - We prospectively studied 712 consecutive patients during a 1-year period who presented with amnesia or loss of consciousness after nonpenetrating head trauma and who had a perfect Glasgow Coma Scale score of 15. Of the 67 (9.4%) patients with acute traumatic lesions disclosed by computed tomography (CT) of the head, 2 required neurosurgical intervention and 1 died. Four factors were statistically correlated (P < 0.05) with abnormal CT findings: Older age, white race, signs of basilar skull fracture, and being either a pedestrian hit by a motor vehicle or a victim of an assault. Sex, length of antero- or retrograde amnesia, forward and reverse digit spans, object recall, focal abnormality on the general neurological exam, and subjective complaints were not statistically correlated with CT abnormality. Using step-wise discriminant function analysis, no single item or combination of items could be used to classify 95% of the patients into either the normal or abnormal CT group. Therefore, regardless of age, mechanism of injury, or clinical findings, intracranial lesions cannot be completely excluded clinically on head-trauma patients who have loss of consciousness or amnesia, even if the Glasgow Coma Scale score is 15. However, only two patients (0.3%) required neurosurgical intervention. PMID- 8421562 TI - Human brain tumor cyst fluid is mitogenic for primary astrocytes in culture. AB - Tumor cyst fluid from five patients with malignant glial neoplasms was assayed for mitogenic activity by measuring [3H]thymidine uptake by rat astrocyte cultures in serum-free defined media. Cyst fluid from all patients stimulated [3H]thymidine uptake three- to fourfold in astrocyte cultures in comparison with untreated controls. Mitogenic activity was found to be soluble and resistant to freezing, but inactivated by heat and trypsin. The activity was retained by ultrafiltration through a 100-kd molecular weight filter. It appears from these results that mitogenic factor(s) for nonneoplastic astrocytes are present in human brain tumor cyst fluid and that such factors appear to be protein in nature and associated with a complex of molecular weight greater than 100,000. PMID- 8421563 TI - Poised for the 21st century. PMID- 8421564 TI - Faculty practice: criterion for academic advancement. PMID- 8421565 TI - MSN in nursing administration and the dual degree. PMID- 8421567 TI - Why men choose nursing. PMID- 8421566 TI - Pay equity and North American nurses. PMID- 8421568 TI - Pamela Maraldo: a bias for action. PMID- 8421569 TI - AIDS: advocacy and activism. Dear President-elect Clinton. PMID- 8421571 TI - Providing cast care. PMID- 8421570 TI - Ensuring patient compliance. PMID- 8421572 TI - Levoxine order could be misread. PMID- 8421573 TI - Reviewing clinical skills, Part III. PMID- 8421574 TI - Recognizing benzodiazepine overdose. PMID- 8421575 TI - Viral hepatitis: unscrambling the alphabet. PMID- 8421576 TI - Low pulse: sign of toxicity? PMID- 8421577 TI - It's okay to die. PMID- 8421578 TI - Postoperative confusion: helping your patient emerge from the shadows. PMID- 8421579 TI - Interpreting C.P.K. & L.D.H. results. PMID- 8421580 TI - How to access an implanted port. PMID- 8421581 TI - John had AIDS--and one romantic wish. PMID- 8421582 TI - Restoring speech with tracheoesophageal puncture. PMID- 8421583 TI - 5 goals for managing older patients. PMID- 8421585 TI - Specialization: the choice is yours. PMID- 8421584 TI - Who's to judge? PMID- 8421586 TI - Using the media to improve nursing's image. PMID- 8421587 TI - Unselfish love. PMID- 8421588 TI - Defining the differences. PMID- 8421589 TI - High-speed dental handpieces and spread of airborne infections. AB - Demonstration of a suction effect in operation of high-speed dental handpieces adds credence to concept that the handpiece can be contaminated internally; it also demonstrates a method by which bacteria can be incorporated into the aerosol cloud that occurs when the high-speed drill is used. PMID- 8421590 TI - Establishing the horizontal plane for positioning of the plane of occlusion. AB - The positioning of the teeth according to the proper occlusal plane provides a considerable advantage in fabricating prosthetic appliances and contributes to optimal appearance and function for patients with complete or partial loss of their teeth. A special device simplifies the technique of establishing the horizontal reference for reproducing the occlusal plane. PMID- 8421591 TI - Buyer beware. When it comes to choosing a computer system. AB - Picking a computer system and developing that system is a process not an event. This article outlines the benefits and the pitfalls inherent in the process. PMID- 8421592 TI - Oral side effects of drugs. PMID- 8421593 TI - The path to public service. PMID- 8421594 TI - Chemoradiotherapy of anal carcinoma: tumour response and acute toxicity. AB - This study was performed to evaluate local tumour control and toxicity of chemoradiotherapy given to patients with primary or recurrent carcinoma of the anal canal. A total of 117 patients were admitted to the Norwegian Radium Hospital during the period of 1983-1989, of which 106 received a combination of radiotherapy (50 Gy to the pelvis) and chemotherapy (mitomycin C + 5 fluorouracil). Sixty-five percent of the patients with primary carcinomas presented with advanced tumours (T3 or T4). Good local tumor control was obtained as only 25% of the patients with advanced tumours (T3 and T4) and 7% of those with smaller tumours (T1 and T2) had a local relapse after treatment. Recurrent tumours following primary surgery did not respond as well; 50% of these patients still had carcinoma 1 month following therapy. All patients experienced acute toxicity (dermatitis/mucositis, diarrhoea and general fatigue), and 50% needed as split course. There was, however, no therapy-related mortality. Survival data seem promising, but further follow-up is necessary to make conclusions. Eight percent of the patients had serious anal insufficiency after treatment. We conclude that the present regimen provides good local tumour control and well preserved anal function, but has considerable acute toxicity. PMID- 8421595 TI - Chemonecrosis for localized dynamic destruction of hepatic metastases of colonic cancer. A new approach. AB - Injection of ethyl alcohol in high concentrations into tissues produces coagulative necrosis. The benefits of direct injection of 98% ethanol into hepatic metastases from 1,2-dimethylhydrazine (DMH)-induced colonic cancer was investigated in groups of 20 Sprague-Dawley rats of either sex. At 10 weeks of age, rats were subcutaneously injected every week with 10 mg/kg DMH for 28 weeks. They were then housed for 3 months. At the end of this period all the animals had developed colonic carcinoma with multiple hepatic metastases. Total colectomy and the fashioning of an ileostomy coupled with direct injection of 0.1-0.2 ml of ethanol into each of the hepatic metastases, after mobilizing the liver by dividing its fascial attachments to facilitate easier tumour detection by inspection and palpation, afforded a significant survival advantage relative to colectomy alone (20.1 +/- 0.2 months of age, vs, 12.8 +/- 0.2 months of age, mean +/- SEM, n = 20, p < 0.01 by the Mann-Whitney U test). The clinical implications of these observations were, therefore, examined in a pilot study carried out on 6 patients (2 women and 4 men with an age range of 43-71 years, mean 56) who had adenocarcinoma of the sigmoid colon with multiple hepatic secondaries. The colonic tumour was resected and an end-to-end anastomosis effected, then all palpable hepatic metastases were slowly injected with 1-1.5 ml of 98% ethanol. Two weeks post-operatively and thereafter once every 2 months any hepatic lesions detected ultrasonically were similarly treated percutaneously. This treatment produced no adverse effects of any significance. None of the patients died during the first post-operative year. Death was due to disease spread in all the patients. The mean survival measured from the time of tumour resection until death from any cause was 20 months (range 17-26 months). It thus appears that chemonecrosis for the treatment of liver metastases from colonic cancer is a simple and safe therapeutic modality which offers a survival gain. PMID- 8421596 TI - Assay, isolation and characterization of circulating immune complexes from serum of gastrointestinal cancer, stage III and IV melanoma and chronic inflammatory bowel disease patients. AB - Circulating immune complexes (CIC) have been detected in several autoimmune diseases, and studies have also suggested that CIC provide a useful tool as tumor markers. In order to identify differences or similarities in antigenic composition, CIC from 23 patients with gastrointestinal (GI) tumors, from 20 patients with stage III and IV melanoma and from 6 patients with inflammatory bowel disease (IBD) were studied. Serum from all GI, melanoma and IBD patients showed higher levels of CIC than controls. SDS/PAGE electrophoresis under reducing conditions revealed some differences between cancer and IBD patients as far as the CIC protein composition was concerned. In melanoma patients, two fast migrating bands, in the regions of 71-74 and 30-49 kD, were found, consistent with previously isolated and characterized antigens described in the literature. PMID- 8421597 TI - DNA ploidy of colorectal carcinoma by tumour site, gender and history of noncolorectal malignancies. AB - DNA ploidy was analysed by flow cytometer from frozen samples of 205 colorectal carcinomas. Sixty-two percent of the tumours had an abnormal DNA stemline. Forty eight percent of carcinomas in the right colon, 62% of carcinomas in the left colon and 74% of carcinomas in the rectum were aneuploid (p = 0.007). Sixty-nine percent of tumours in males and 55% of tumours in females were aneuploid (p = 0.029). The difference in frequency of aneuploidy between females and males was greatest in tumours of the right colon, where 37% of the tumours in females and 62% of the tumours in males were aneuploid (p = 0.047). The percentage of diploid colorectal carcinomas was higher (55%) in patients with a history of noncolorectal malignancy than in others (34%, p = 0.031). These results suggest that flow cytometry may be helpful in understanding the development of colorectal carcinomas. PMID- 8421598 TI - Usefulness of nucleolar organizer region staining in gastric myogenic tumors: correlation with ploidy by DNA flow cytometry. AB - A silver-staining technique to identify nucleolar organizer region-associated proteins (Ag-NOR) was applied to 55 gastric myogenic tumors. The mean numbers of Ag-NOR in the nucleus were: leiomyoma (30 cases), 2.0 +/- 0.6 (mean +/- SD); low grade leiomyosarcoma (11 cases), 3.0 +/- 0.7; and high-grade leiomyosarcoma (14 cases), 3.9 +/- 0.7. The Ag-NOR counts were compared with DNA ploidy as determined by flow cytometry in 46 tumors. The Ag-NOR counts were significantly different in the aneuploid leiomyosarcomas (4.1 +/- 0.6: mean +/- SD) and diploid leiomyosarcomas (2.9 +/- 0.6: mean +/- SD). Thus, this parameter may serve as an objective histological discriminant for malignancy. PMID- 8421599 TI - Predictive value of a morphometric prognostic index in female breast cancer. AB - Survival data of the patients were correlated to tumour size, axillary lymph node (pN) status, mitotic frequency and morphometric prognostic index (MPI) in a series of 611 women with a primary breast carcinoma treated and followed-up for over 12 years in Kuopio University Hospital. The pN status, tumour size, mitotic activity index (MAI), volume-corrected mitotic index (M/V index) and MPI all predicted recurrence-free survival and cancer survival (p < 0.001). In pN(-) patients, the MPI was the most important predictor of recurrence-free survival and cancer survival (p < 0.001) followed by the mitotic frequency. In pN(+) patients, tumour diameter and MPI were equal predictors (p < 0.001) of survival followed by M/V index. In Cox's analysis, MPI, pN status and mitotic frequency independently predicted survival in the whole series. In the separate analysis of pN(-) and in pN(+) tumours, the MPI and MAI independently predicted survival. The M/V index was independently related to recurrence-free survival in pN(+) tumours. In multivariate analysis, MPI was an independent predictor although it does not include all the prognostic information. The results suggest that the decisions on adjuvant therapy in breast cancer can be based on the MPI, particularly in pN(-) patients. PMID- 8421600 TI - Growth inhibition and modulation of cell markers of melanoma by S-allyl cysteine. AB - A sulfur-containing amino acid compound, S-allyl cysteine (SAC), derived from garlic extract inhibited proliferation of nine human and murine melanoma cell line in a dose-dependent manner (1.2-10 mM) assessed by a [3H]thymidine incorporation assay. Three control human lymphoblastoid cell lines were not inhibited by SAC concentrations < 5 mM. Four human melanoma cell lines in a soft agar assay also showed dose-dependent inhibition of colony formation by SAC. Melanin content was increased up to 95% compared to the same untreated cell lines in these four human melanoma and two B16 murine melanoma sublines. Expression of cell surface gangliosides, cellular-differentiation and transformation markers, decreased after SAC treatment. Significant morphological changes including 'flattening and/or dendritic-like elongations' were also observed. Thus SAC inhibited cellular growth and proliferation and modulated major cell differentiation markers of melanoma. PMID- 8421601 TI - Position specificity of Ki-ras oncogene mutations during the progression of colorectal carcinoma. AB - The Ki-ras proto-oncogene is converted into an active oncogene by mutations in codon 12, 13, or 61. The incidence of mutations in the Ki-ras oncogene in colorectal adenomas and primary colorectal carcinomas has been shown to be 50-75 and 40-65%, respectively. To determine the role activation of the Ki-ras oncogene plays in the progression of colorectal carcinoma, we analyzed DNA from 11 nude mouse xenografts and from 24 metastases of 22 patients with colorectal carcinoma, using the polymerase chain reaction technique and hybridization with labeled mutation-specific oligomers. Eleven of the 24 metastases (46%) carried mutations, 7 in codon 12 and 4 in codon 13, whereas only 1 nude-mouse tumor (9%) harbored a Ki-ras codon-12 mutation. Eleven of these 12 mutations in advanced stages of colorectal cancer were localized to the second position of either codon 12 or codon 13, whereas a majority of published ras mutations in earlier stages are in the first position of codon 12 of the Ki-ras oncogene. We conclude that there is a position specificity of Ki-ras oncogene mutations in advanced stages of colorectal carcinoma. In general, however, these mutations do not seem to play an important role in the progression of this cancer. PMID- 8421602 TI - A historical review of shoulder arthroscopy. AB - The last decade has seen a huge increase in the application of arthroscopic techniques to the shoulder. From early efforts around 1980, arthroscopic shoulder surgeries have become well established and accepted. With the creative development and application of new techniques, the scope of shoulder pathology amenable to arthroscopic treatment continues to expand. PMID- 8421603 TI - Arthroscopic treatment of synovial diseases of the shoulder. AB - A wide spectrum of synovial disorders may affect the shoulder joint. This article provides a detailed description of the clinical features of synovial disorders of the shoulder. The role of arthroscopic surgery in the diagnosis and treatment of this spectrum of disease is discussed also. PMID- 8421604 TI - Arthroscopic debridement and decompression for selected rotator cuff tears. Clinical results, pathomechanics, and patient selection based on biomechanical parameters. AB - Clinical results of arthroscopic rotator cuff debridement and decompression in 25 patients are presented. Additionally, a fluoroscopic study of 14 patients with known massive rotator cuff tears reveals four kinematic patterns in patients with major cuff deficiencies. Biomechanical models are used to support further the clinical impression that certain rotator cuff tears are amenable to arthroscopic debridement and decompression without repair. Specific indications for arthroscopic debridement and decompression are developed and presented. PMID- 8421605 TI - Management of rotator cuff calcifications. AB - Although investigators do not agree on the etiology of calcification of the rotator cuff, it may be linked to hypoxia of the tissue. New evidence suggests that there may be a genetic predisposition linked to the HLA-A1 antigen. The initial phases of formation of the calcification are rarely symptomatic. The acute phase symptoms that debilitate the patient are usually associated with the resorptive phase, in which there is vascular invasion and influx of phagocytic cells, increasing the intratendinous pressure and exacerbating the symptoms. Conservative treatment including local injections of anesthetic, needling, and barbotage is frequently successful. Steroid injections are controversial and may slow the long-term resorption of calcium. A small group of patients remain symptomatic. For these patients and for the chronic subacute patient who fails to resolve with conservative treatment, excision of calcium offers reliable relief. Previous experience with open excision provided predictable results but with a surprisingly long time to recovery. Recent experiences with arthroscopic excision have decreased the morbidity, and several investigators have reported uniformly excellent results. The technique is demanding, but arthroscopy permits reliable removal of the calcification and resolution of pain. Acromioplasty with or without coracoacromial ligament resection should be performed only in patients in whom impingement has been demonstrated by physical examination or intraoperative arthroscopic examination. PMID- 8421606 TI - Arthroscopic resection of the distal clavicle. AB - Painful conditions of the acromioclavicular joint without instability can be treated successfully with arthroscopic methods. The direct approach is best suited for isolated acromioclavicular pathology. It also can be used to address the acromioclavicular joint during shoulder arthroscopy and bursoscopy, but two additional acromioclavicular portals are needed. In patients with both subacromial and acromioclavicular joint pathology, the bursal approach to the acromioclavicular joint can be used. In some patients with narrow or medially inclined overriding clavicles, the distal clavicle is not easily resected with the bursal approach. The direct approach is an alternative in these situations. Either method has been shown to be an effective treatment and can return the patient to full activity much sooner than with a traditional open resection. PMID- 8421607 TI - Our technique for the arthroscopic Mumford procedure. AB - This article describes a new technique and instrumentation for performing arthroscopic distal clavicle resection safely and accurately using the three standard portals for shoulder arthroscopy. A simple six-step surgical technique is reviewed. Clinical results of patients who have undergone an arthroscopic Mumford procedure using this technique are presented also. PMID- 8421608 TI - Arthroscopic acromioplasty. The superiority of the posterior portal over the lateral portal. AB - The authors demonstrate that the posterior portal is superior to the lateral portal for performing an arthroscopic acromioplasty by using pre- and postoperative three-dimensional computed tomography scans to quantitate bone shape and volume. The posterior portal gives superior resection of volume of bone as well as superior contour of the bone, especially in removing the anterior acromial hook. PMID- 8421609 TI - Arthroscopic subacromial decompression and postoperative management. AB - Arthroscopic subacromial decompression is the arthroscopic equivalent of a standard open procedure. Although technically demanding, it facilitates early rehabilitation. The results in patients with stage II disease are equal or better than those achieved through open surgery. The procedure is also useful in selected patients with stage III disease in whom pain is the major complaint. Complete rotator cuff tears can be repaired into a bony trough through a small deltoid-splitting incision. PMID- 8421610 TI - Evaluation and treatment of the rotator cuff. AB - The use of the arthroscope for evaluation and treatment of rotator cuff problems is becoming more common. Technology has advanced to the point where we can now not only positively diagnose early rotator cuff problems, but successfully treat them; many times without open shoulder surgery. Arthroscopic cuff debridement and decompression are the mainstays of surgical prevention. Arthroscopic suture fixation of delaminated cuff tears or small rotator cuff avulsions is now possible. The miniopen rotator cuff approach is excellent for repairing most larger full-thickness tears. This article reviews all of these current topics and offers step-by-step surgical techniques used and recommended by the author. PMID- 8421611 TI - Arthroscopic portals: anatomy at risk. AB - Proper portal placement is critical for successful and safe arthroscopic surgery. Multiple portals and their safe location are needed for evolving surgical procedures. Familiarity with these portals and their formation will aid the arthroscopist. PMID- 8421612 TI - Office arthroscopy of the shoulder. A diagnostic alternative. AB - Office arthroscopy of the shoulder is a technically demanding procedure and is significantly more challenging than office arthroscopy of the knee. With the proper equipment and in the hands of an experienced shoulder arthroscopist, the procedure offers many advantages. Patients unanimously prefer office arthroscopy to MR imaging because of the ability to view their pathology personally in a medium more palatable and understandable than MR imaging. Office arthroscopy appears to be a safe, accurate, and cost-effective alternative to MR imaging for the diagnosis of shoulder pathology in select patients. PMID- 8421613 TI - Arthroscopic anatomy of the glenohumeral joint and subacromial bursa. AB - A review of normal glenohumeral and subacromial anatomy and the clinical relevance of normal variants and pathologic conditions has been presented. This information will aid the arthroscopist in making accurate diagnoses and giving effective treatment. PMID- 8421614 TI - Evaluation and treatment of biceps tendon pathology. AB - Until recently, the role of the long head of the biceps tendon as a source of shoulder pain had been controversial. With careful examination, improved imaging techniques, and arthroscopy, a specific diagnosis can be made. In 95% of patients, biceps tendinitis is secondary to a primary diagnosis of impingement syndrome. Subluxation of the biceps tendon and primary biceps tendinitis are diagnoses of exclusion. However, in the properly selected patient, both respond well to biceps tenodesis. Rupture of the long head of the biceps tendon can be the end result of any of these pathologic processes. In the older sedate patient, conservative treatment results in little functional loss and a mild cosmetic deformity. In the young active patient, especially those who perform tasks that require supination strength, a primary biceps tenodesis should be performed, as well as decompression if there is any evidence of impingement. The key to successful treatment of lesions of the long head of the biceps tendon is recognition of associated pathologic findings in the shoulder. With advances in arthroscopy, the orthopedist can tailor treatment exactly to the pathology, minimizing morbidity and maximizing a successful outcome. PMID- 8421615 TI - Injuries to the glenoid labrum, including slap lesions. AB - The glenoid labrum of the shoulder has extensive anatomic variation but appears to be important for contributing to shoulder stability and for increasing the depth of contact between the glenoid labrum and the humeral head. Tears of the labrum are commonly seen in association with other pathologic entities, such as instability and rotator cuff tears, and treatment of the labral pathology may be incidental to treatment of the other more significant pathology. However, conditions isolated to the labrum do occur and can be a significant source of shoulder problems. Effective treatment of these lesions may result in significant improvement in the patient's symptoms. Labral lesions are difficult to diagnose, and special diagnostic studies and, frequently, arthroscopy are required. The recently described SLAP lesion is an uncommon but significant cause of shoulder disability that generally requires arthroscopic diagnosis. The arthroscopic treatment of this lesion depends on the type of SLAP lesion present. Recent techniques have permitted arthroscopic stabilization of the biceps labral detachment and type II SLAP lesions. PMID- 8421616 TI - Arthroscopy. Basic setup and equipment. AB - Shoulder arthroscopy necessitates attention to detail. Adopting a comfortable setup that includes a plan for every detail, including the surgeon's position, the patient's position, the draping techniques, and the types of fluid and equipment necessary, adds to the smooth flow of the operation even when the surgical plan changes. I have described my method of basic arthroscopic setup and equipment, which has helped me to deal with the many frustrations faced during arthroscopy on the shoulder. It is hoped this article may enable the reader to avoid some of these problems if this or a similar basic setup is adopted. PMID- 8421617 TI - The rationale and technique for arthroscopic reconstruction of anterior shoulder instability using multiple sutures. AB - Recent anatomic and arthroscopic studies have increased knowledge of the pathology of traumatic anterior shoulder instability. Advances in arthroscopy have made reconstruction of the shoulder using this technique seem increasingly attractive. The authors review the rationale and technique of arthroscopic reconstruction with multiple sutures. PMID- 8421618 TI - Arthroscopic capsulolabral repair using suture anchors. AB - This article presents a different arthroscopic approach to the diagnosis and treatment of anterior shoulder instability. Surgical technique, the postoperative regimen, and the results and pitfalls of surgery using suture anchors are discussed. PMID- 8421619 TI - Arthroscopic shoulder capsulorrhaphy using metal staples. AB - Staple capsulorrhaphy on the shoulder using a metal staple for traumatic anterior instability has the advantages of increased diagnostic accuracy, microdebridement of the pathology, accurate assessment of the glenohumeral ligament pathology, and selective repair of the ligament pathology. Although the same advantages should apply to staple capsulorrhaphy for traumatic posterior instability, our experience remains very limited. Staple capsulorrhaphy on the shoulder has multiple disadvantages, including being technically difficult with a slow learning curve, not being applicable to all unstable shoulders, an average failure rate of 12% that may be related to inadequate postoperative immobilization, no extra-articular reinforcement, and the use of a metal implant that may need to be removed at a second operation. Staple capsulorrhaphy is currently performed for traumatic anterior instability in the shoulder, with a selected repair of the pathology using a single, well-placed staple and prolonged postoperative immobilization. The design of the staple affords a simpler insertion technique than rivets, screws, and intra-articular sutures. The advent of a biodegradable staple should eliminate inherent problems of metal implants while preserving the advantages of this method. PMID- 8421620 TI - Instability versus impingement syndrome in the throwing athlete. AB - Shoulder pain in the throwing athlete has become a well recognized phenomenon. Shoulder pain in the athlete is often due to overuse and typically responds with a well managed conservative program that includes rest and rotator cuff rehabilitation exercises. The author's experience has demonstrated that persistent pain and symptoms of an impingement syndrome are often due to an unrecognized instability syndrome in the athlete. PMID- 8421621 TI - [Diagnosis and therapy of retroperitoneal tumors]. AB - Upon detecting 112 retroperitoneal space reducing processes the authors operated on 62 adults with primary retroperitoneal tumours. Of them there were 12 benign and 50 malignant tumours. The diagnostics and complex therapy of these tumours are dealt with. It is established that although the CR, MR, and USG have resulted in a considerable development the therapeutic results are still not far better. In the surgical management of benign lesions total tumour exstirpation is recommended even with extensive multivisceral resections. For tumours of low grade malignancy the authors have the same standpoint. In case of retroperitoneal tumours of high grade malignancy--and a considerable number of patients belongs to this group--the ultraradical interventions are not associated with promising results therefore they are not recommended by the authors. In the complex management of primary retroperitoneal tumours the achievements of radio- and chemotherapy are nowadays rather only promising than notable. PMID- 8421622 TI - [Effect of acipimox on diabetes mellitus-associated hyperlipoproteinemia]. AB - The authors give summary on the pathogenesis of hyperlipoproteinaemia in patients with diabetes mellitus. They investigated the effects of acid nicotinic derivate acipimox on lipid metabolism in these patients. They treated with acipimox 15 patients who suffered from non-insulin-dependent diabetes mellitus, as well as II/B, IV type hyperlipoproteinaemia by Fredrickson. They investigated the effect of acipimox on the lipoprotein content and quality in the sera and uric acid and carbohydrate metabolism. The authors found decreased cholesterol, triglyceride, apolipoprotein-B, glycosilated haemoglobin, glucose and uric acid levels in the sera and they found that the atherogenic index decreased too. During the treatment they could not recognise a change in the low-density lipoprotein level of the sera while the high density lipoprotein and apo-A1 level increased. PMID- 8421623 TI - [Autologous spleen transplantation]. AB - Splenectomy is known to increase the risk of overwhelming bacterial infection. There is a decrease in immunoglobulin IgM, T-lymphocytes, impaired primary antibody response to antigen challenge, an altered opsonic function and a tuftsin deficiency. Splenic autotransplantation has been suggested as a method of preserving function and this concept is supported by experiments in animals (dogs). The study describes autotransplantation of the traumatized spleen in human beings for the preservation of splenic function. Eleven patients operated on for abdominal trauma in the Kenezy Hospital in Debrecen, required total splenectomy, than splenic autotransplantation. In these patients splenic slides were implanted in between two layers of omental pouch (Furka's "spleen chip"). In 10 patients the follow-up radionuclid imaging, the IgM level, and the tuftsin level unambiguously confirmed the functioning of the splenic tissue. PMID- 8421624 TI - [Genital chlamydia infection in andrologic patients. Direct antigen detection and serologic screening]. AB - Authors investigated the frequency of genital chlamydial infection. 83 couples- complaining with infertility--were examined. Twenty patients were chlamydia positive by direct IF antigen detection. All chlamydia positive patients have elevated IgG antibody levels in sera. In 33 cases (18.1%) the antibody titers were significantly high above 1:128, in 58 cases (34.9%) antibody levels were slightly elevated between 1:16-1:64. Authors are analysing the questions of the humoral immune reactions and the results of the medical treatment. PMID- 8421625 TI - [Pathogenic role of a pacemaker in occult infection]. AB - A case of seronegative pacemaker lead infection is presented. The diagnosis based on patient history and echocardiography was proved by removing the infected leads from the beating heart through surgery. PMID- 8421626 TI - [Reorganization of the Hungarian public health system]. PMID- 8421627 TI - Dislocation of the sternoclavicular joint. Evaluation using paraxial computed tomographic reconstruction. AB - Sternoclavicular dislocations are relatively infrequent, constituting less than 1% of somatic dislocations. Despite the fact that the sternoclavicular joint is the only articulation between the upper extremity and the axial skeleton, it possesses the least amount of osseous stability of any joint in the body. Sternoclavicular dislocations are generally divided into anterior and posterior disruptions, the former being the most common. An unusual case of an anterior dislocation of the sternoclavicular joint with a large superior component is described. It was found that coronal paraxial computed tomographic reconstruction of the joint was quite useful in evaluating this injury. PMID- 8421628 TI - A 19-month-old female with unilateral bowed leg. AB - The following case is presented to illustrate the roentgenographic findings of a condition of interest to the radiologist and orthopaedic surgeon. Initial history, physical findings, and roentgenographic examinations are presented, along with the final histologic and roentgenographic differential diagnosis. PMID- 8421629 TI - A new use of the Herbert bone screw system: treatment of a displaced radial styloid fracture. AB - A 28-year-old man sustained a displaced radial styloid fracture. Roentgenograms revealed a significant intra-articular step-off. The Herbert Bone Screw System was used to obtain an anatomic reduction. This method of treatment for a displaced radial styloid fracture has not been previously reported. PMID- 8421630 TI - The "flatback" cast. AB - A properly applied above-elbow cast in a child presents difficulties not seen in adults. Because of the differences in anatomy, physiology, and compliance, children can benefit from modification of the standard above-elbow cast. We report on a method of application of the "flatback" cast to address these concerns. PMID- 8421631 TI - Shoulder dislocation. PMID- 8421632 TI - Big Red is back. PMID- 8421633 TI - Pedicle screw fixation in the management of unstable thoracolumbar spine injuries. PMID- 8421634 TI - Medical therapy of low back pain. AB - Low back pain is a common clinical symptom usually associated with mechanical disorders, primarily muscle strain, affecting the lumbosacral spine. The majority of patients with low back pain improve within a 2-month period with conservative medical management. Conservative management includes limited physical activity, injections, nonsteroidal anti-inflammatory drugs, and muscle relaxants. These therapies work best in combination and are very effective in decreasing low back pain. PMID- 8421635 TI - Variations of the ulnar nerve. A surgical perspective. AB - A systematic review of ulnar nerve variations is presented. Many of these anomalous neural structures account for the atypical clinical or electromyographic findings that are often a source of diagnostic confusion. Knowledge of these variations will hopefully lessen the likelihood of inadvertent injury and consequent motor and/or sensory loss. PMID- 8421636 TI - Internal fixation of proximal femur fractures: a biomechanical comparison of the Gamma Locking Nail and the Omega Compression Hip Screw. AB - The fracture fixation capability of the Gamma Locking Nail was compared with the Omega Compression Hip Screw using cadaver femurs with simulated intertrochanteric and subtrochanteric fractures. Loading of the femurs was done on an Instron machine. Fracture fragment motion was recorded at 100-lb loading increments using standard roentgenograms. Both forms of hardware exhibited sufficient structural integrity to carry loads up to 600 lbs. Fracture motion was generally greater with the Gamma Locking Nail, which appeared to suspend the proximal fragments above a pivot point defined by the distal locking screws. Compression of the fracture site under loading did not occur with the Gamma Locking Nail. The potential for iatrogenic fracture generation appeared greater with the Gamma Locking Nail. Additionally, no clear mechanical advantage was documented in the fracture patterns tested. A limited discretionary use of the Gamma Locking Nail is advised. PMID- 8421637 TI - Treatment of rotator cuff impingement. AB - The author performed 497 shoulder decompression procedures between 1978 and 1989. Of these procedures, 252 (51%) had tears of the rotator cuff that were repaired concomitantly. The results were successful in over 90% of the cases. It is suggested that articular-surface tears or intratendinous tears are the possible cause of some of the failures of the open procedure. The findings also suggest that the use of arthroscopy or magnetic resonance imaging may be indicated in selected cases. However, for the appropriate patient, open shoulder decompression continues to be an excellent treatment. PMID- 8421638 TI - Results of endoscopic management of carpal tunnel syndrome. AB - This paper is a retrospective study of 27 women with carpal tunnel syndrome (39 hands) who underwent a new endoscopic operative procedure utilizing the Universal Subcutaneous Endoscope system developed by the lead author. Operations on 199 hands were performed under local anesthesia on an outpatient basis. The etiology was considered idiopathic in all cases. Complete preoperative and postoperative clinical and electrophysiologic data were analyzed in 39 hands. The follow-up period ranged from 12 to 45.8 months (mean, 18.3 months). Symptoms of sensory disturbances disappeared in an average of 20 weeks in all patients. Electrophysiologic studies showed definite improvement when compared with preoperative studies. No complications were recorded. Clinical and electrophysiologic results showed that the less-invasive management of carpal tunnel syndrome by endoscopy is safe and effective. PMID- 8421639 TI - High radial nerve palsy in a tennis player. AB - High radial nerve palsy occurred in a 47-year-old athlete following muscular overexertion. Surgical exploration showed a fibrous arch coming from the long head of the triceps and causing entrapment of the radial nerve. Radial nerve entrapment following muscular overexertion has been reported to be caused by fibrous arches coming from the lateral head of the triceps. Fibrous arches can occur from either the lateral or long head of the triceps muscle. The neuropathy that occurs can be irreversible. PMID- 8421640 TI - Use of the Herbert bone screw compression jig to reduce and stabilize a Bennett fracture. AB - This case report describes a new use for the compression jig from a Herbert bone screw set. The use of this device in stabilizing a Bennett fracture of the thumb metacarpal prevented extensive palmar exposure and facilitated reduction of the fracture. PMID- 8421641 TI - USP institutes new mandates for potassium chloride for injection concentrate. PMID- 8421642 TI - Development of a vital-sign/fluid-balance flow sheet. AB - An improved flow sheet for recording vital signs and fluid balance on a medical oncology unit was developed and tested using quality-assurance techniques. The new form, which replaced three separate forms, measurably improved documentation on all quality-assurance monitors tested. Additional benefits include cost savings and decreased time expenditures by nursing staff. PMID- 8421643 TI - Card provides quick access to outpatient chemotherapy information. PMID- 8421644 TI - Ingenuity reaps results in the homecare setting. PMID- 8421645 TI - Interinstitution coordination smooths transitions. PMID- 8421646 TI - Planning guide increases patient referrals. PMID- 8421647 TI - Form enhances communication within hospital system. PMID- 8421648 TI - Causal attribution, perceived control, and adjustment in patients with lung cancer. AB - The relationships among causal attribution, perceived control, and adjustment to lung cancer were examined in 61 outpatients who had received a diagnosis of primary lung cancer. Data were collected using a structured interview and a self report questionnaire. Both internal and external causal attributions were significantly positively correlated with perceived control. The relationship between internal causal attribution and perceived control was stronger. No significant relationships were found between perceived control and adjustment, although both internal and external causal attributions were significantly negatively correlated with aspects of adjustment. Recommendations are made for future research. PMID- 8421650 TI - Acute tumor lysis syndrome: assessment and nursing implications. AB - Acute tumor lysis syndrome (TLS) is a metabolic complication that occurs in some patients with cancer and frequently is triggered by chemotherapy. TLS results from a rapid destruction of a large number of tumor cells, which causes several potentially fatal biochemical changes. If not identified and treated quickly, TLS can result in acute renal failure. TLS often is associated with tumors that have high growth fractions. Typically, these tumors are very sensitive to the effects of chemotherapeutic agents. Fortunately, this complication can be prevented with appropriate medical and nursing management of patients at high risk. This article reviews the pathogenesis and medical treatment of TLS and identifies nursing implications for oncology nursing practice. PMID- 8421649 TI - The effect of preparation for lumbar puncture on children undergoing chemotherapy. AB - At the University Hospital in Lund, Sweden, a preparation program was developed for children undergoing lumbar punctures (LPs) during chemotherapy for leukemia or lymphoma. Subsequently, a study was initiated to determine whether a preparation program for children prior to treatment would reduce their anxiety and improve their cooperation. This study also was undertaken to examine whether reinforcing the preparation information prior to each LP would be beneficial. The 30 children who participated in the study were divided into three groups: a control group whose members did not receive preparation and two other groups whose members were exposed to different numbers of preparation programs. The parents and the nurse in charge evaluated the children's reactions during treatment using two 6-point rating scales: an anxiety scale and a noncooperation behavior scale. Two unbiased, trained observers later viewed video recordings of the children's reactions and evaluated them using the same tools. In addition, each child rated his or her experience of pain on a 10 cm visual analogue scale. Based on these ratings, the groups were analyzed to determine if within-group differences existed from one treatment to the next and to determine if between group differences existed at the various times of treatment. Few statistically significant differences were found, but the results indicate that the children in the most informed group exhibited sustained reductions in their perceptions of pain. This may signify that reinforcing the preparation information before each of the LPs enabled these children to cope with the pain more effectively. PMID- 8421651 TI - Intravenous immunoglobulin in oncology nursing practice. AB - Intravenous immunoglobulin (IVIG) is a concentrated form of IgG, also known as gamma globulin, that is derived from the pooled serum of a large number of donors. IVIG contains many types of antibacterial and antiviral antibodies. While its use in certain clinical conditions (e.g., severe combined immunodeficiency) is well-established, other indications still are under investigation. Along with nursing implications for use in inpatient and outpatient settings, the role of IVIG in treating immune thrombocytopenic purpura, chronic lymphocytic leukemia, treatment-induced neutropenia and thrombocytopenia, bone marrow transplantation, and AIDS will be discussed. PMID- 8421652 TI - Comparison of perceived symptoms of patients undergoing bone marrow transplant and the nurses caring for them. AB - This study describes the symptomatology of patients hospitalized for bone marrow transplant (BMT) (n = 30) and compares their perceptions of these symptoms to those of nurses (n = 28). Patients and nurses responded to the Symptom Distress Scale (SDS) four times: within 48 hours of BMT day one (T1); day 7-10 post-BMT (T2); day 20-23 post-BMT (T3); and day 30-34 post-BMT (T4). Patients also completed the Profile of Mood States at T1. Each time, the investigators completed a Karnofsky Performance Status evaluation of each patient. Patients perceived significantly more distress from their symptoms at T1 than their nurses perceived that they were experiencing. Over time, patients' SDS scores did not change significantly. However, nurses' SDS scores indicated significant differences, with their SDS scores at T1 less than those at T2 and T3. The results indicate the importance of nurses exploring the perceived symptom experiences of patients undergoing BMT. Any incongruence between nurses' and patients' perceptions potentially could prevent patients' symptoms from being managed effectively. PMID- 8421653 TI - Thyroid cancer: a review. AB - Thyroid cancer is a rare and complex disease. The thyroid contains various cell types from which distinct diseases arise. These malignancies range from indolent to extremely aggressive. Diagnosis includes attention to risk factors, family history, and subjective reports. The most valuable tool for diagnosis is the fine needle aspiration. Primary treatment is surgery with postoperative hormone therapy. Radiation and chemotherapy serve palliative and adjuvant roles in advanced, recurrent, or metastatic disease. Nurses make a significant contribution to patient understanding and successful treatment outcome. PMID- 8421654 TI - Postradiation sarcoma of bone. PMID- 8421655 TI - The International Documentation and Evaluation System (IDES) PMID- 8421656 TI - Changing patterns of prophylaxis for deep venous thrombosis following elective hip replacement. AB - The author audited the charts of elective hip replacements at a community hospital over an 18 month period. Initially, aspirin was the most commonly ordered prophylactic agent (31%). Only 15% of patients received an ideal or probably effective prophylactic regimen. After feedback was received by the orthopedists, another audit was performed. Warfarin was the most commonly used agent and 54% of patients received an ideal or probably effective regimen in the follow-up period (P < .0001 compared to initial survey). Although there is still room for further improvement, orthopedists in this community are using more effective regimens to prevent thromboembolism following elective hip surgery. PMID- 8421657 TI - Treatment of isolated ulnar shaft fractures with functional bracing. AB - A prospective study was initiated using functional bracing to treat isolated ulnar shaft fractures. Forty-seven consecutive patients with 48 fractures were treated from July 1987 to January 1989. Twenty-eight patients with 29 fractures were available for follow up. Three of these fractures were subsequently excluded, as they had open reduction and internal fixation, leaving a group of 26 fractures. The mean time to union was 45 days (6.4 weeks). Fractures of the middle third of the ulnar diaphysis had the highest mean time to union, as did those with an oblique fracture pattern of 26% to 50% initial displacement. For union time, there was no significant difference for fracture location (P > .05 via t-test) or for fracture pattern (P > .05 via analysis of variance). PMID- 8421658 TI - The medial approach to the hip revisited. AB - Several muscular intervals have been described to approach the hip through a medial incision. We studied in detail the original descriptions of these surgical planes and found that several of them are nearly identical. We attempt to clarify the anatomic intervals available to the surgeon in hope of eliminating ambiguity in future reporting. PMID- 8421659 TI - Displaced proximal humeral fractures. PMID- 8421660 TI - Philosophy of osteosynthesis in shoulder fractures. AB - A wide range of workable instrumentation is available for the treatment of proximal humeral fractures, whereas osteosynthesis of scapular fractures is commonly achieved by plate/screw fixation. Older persons are confronted with a steadily rising incidence of osteoporotic fractures. Especially for this group of patients, minimally invasive and minimally instrumented surgical procedures, such as K-wiring and bone sutures, seem to have superseded plate fixation. For the treatment of scapular fractures, an important question remains: Which fracture types provide better functional results with open reduction and internal fixation, and which with conservative treatment? Our indications for an operative procedure are limited to the floating shoulder, dislocated scapular neck fracture, and dislocated glenoid fracture. PMID- 8421661 TI - Delta shoulder prosthesis for rotator cuff rupture. PMID- 8421662 TI - Stress fracture MRI. PMID- 8421663 TI - Rotator cuff tears in the adolescent. PMID- 8421664 TI - Chronic osteomyelitis of the calcaneus: treatment with a vascularized muscle flap. PMID- 8421665 TI - Long-term follow up of a case of kyphomelic dysplasia. PMID- 8421666 TI - Posterior shoulder dislocation: pitfalls and perils. PMID- 8421667 TI - Chloroplast gene sequences and the study of plant evolution. AB - A large body of sequence data has accumulated for the chloroplast-encoded gene ribulose-1,5-biphosphate carboxylase/oxygenase (rbcL) as the result of a cooperative effort involving many laboratories. The data span all seed plants, including most major lineages from the angiosperms, and as such they provide an unprecedented opportunity to study plant evolutionary history. The full analysis of this large data set poses many problems and opportunities for plant evolutionary biologists and for biostatisticians. PMID- 8421668 TI - Identification of a nerve growth factor- and epidermal growth factor-regulated protein kinase that phosphorylates the protooncogene product c-Fos. AB - Nerve growth factor (NGF) treatment of rat pheochromocytoma (PC12) cells induces the synthesis of the transcription factor c-Fos, which becomes highly phosphorylated relative to that produced as a result of depolarization of the cell. A peptide derived from the carboxyl terminus of c-Fos (residues 359-370, RKGSSSNEPSSD) containing putative phosphorylation sites was used to detect a NGF stimulated Fos kinase. NGF treatment of PC12 cells resulted in a rapid activation of a protein kinase which phosphorylated both the c-Fos peptide and authentic c Fos at its carboxyl terminus. The kinase was selectively activated by NGF and epidermal growth factor but was not induced by depolarization or other agents. The c-Fos peptide was phosphorylated at a serine corresponding to Ser362, a site critically implicated in the capacity of c-Fos to exhibit transrepressive activity [Ofir, R., Dwarki, V. J., Rashid, D. & Verma, I. M. (1990) Nature (London) 348, 80-82)]. The NGF-stimulated Fos kinase may play an important role in regulating the expression and transforming potential of c-Fos. PMID- 8421669 TI - A yeast expression system for human galactose-1-phosphate uridylyltransferase. AB - Galactose-1-phosphate uridylyltransferase (GALT) (UTP: alpha-D-hexose-1-phosphate uridylyltransferase, EC 2.7.7.10) is an essential enzyme of the Leloir pathway of galactose metabolism. Mutations in human GALT are associated with the potentially lethal disorder galactosemia, which affects 1 in 30,000-60,000 live-born infants. Although a number of base substitutions have been identified in the GALT alleles of galactosemia patients, the detailed biochemical impact of these mutations on GALT enzymatic activity remains obscure. Similarly, little is known about the sequence/structure/function relationships for wild-type human GALT. As a first step toward addressing these questions, we have developed a yeast-based expression system for the human enzyme. The wild-type human GALT coding sequence has been introduced into a strain of Saccharomyces cerevisiae that carries a disruption of the GALT-encoding GAL7 gene and, therefore, expresses no endogenous GALT. Transformants were tested for restoration of GALT activity both indirectly, by cell growth on galactose, and directly, by analysis of enzyme activity in cell extracts. The results of both tests were striking; wild-type human GALT functioned in yeast almost as well as the endogenous enzyme. In contrast, cells transformed with either human or yeast GALT sequences engineered to carry a common human GALT mutation, Q188R (changing Gln188 to Arg), exhibited essentially no detectable GALT activity and failed to grow on galactose. Lymphoblasts from patients homozygous for the Q188R mutation similarly exhibited essentially no detectable GALT activity in parallel assays. The results reported here establish the utility of the yeast-based expression system for human GALT and set the stage for more detailed studies of this important enzyme and its role in galactosemia. PMID- 8421670 TI - Rapid, high-level expression of biologically active alpha-trichosanthin in transfected plants by an RNA viral vector. AB - alpha-Trichosanthin, a eukaryotic ribosome-inactivating protein from Trichosanthes kirilowii, inhibits the replication of the human immunodeficiency virus (HIV) in vitro. The alpha-trichosanthin gene was placed under the transcriptional control of a tobamovirus subgenomic promoter in a plant RNA viral vector. Two weeks after inoculation, transfected Nicotiana benthamiana plants accumulated alpha-trichosanthin to levels of at least 2% of total soluble protein. The recombinant alpha-trichosanthin was purified and its structural and biological properties were analyzed. The 23-amino acid signal peptide was recognized by N. benthamiana and the processed enzyme caused a concentration dependent inhibition of protein synthesis in vitro. The high level of heterologous gene expression observed in these studies is due to the unique features of the RNA viral-based transfection system. PMID- 8421671 TI - Genomic structure and chromosomal localization of the human deoxycytidine kinase gene. AB - Deoxycytidine kinase (NTP:deoxycytidine 5'-phosphotransferase, EC 2.7.1.74) is an enzyme that catalyzes phosphorylation of deoxyribonucleosides and a number of nucleoside analogs that are important in antiviral and cancer chemotherapy. Deficiency of this enzyme activity is associated with resistance to these agents, whereas increased enzyme activity is associated with increased activation of such compounds to cytotoxic nucleoside triphosphate derivatives. To characterize the regulation of expression of this gene, we have isolated genomic clones encompassing its entire coding and 5' flanking regions and delineated all the exon/intron boundaries. The gene extends over more than 34 kilobases on chromosome 4 and the coding region is composed of 7 exons ranging in size from 90 to 1544 base pairs (bp). The 5' flanking region is highly G+C-rich and contains four regions that are potential Sp1 binding sites. A 697-bp fragment encompassing 386 bp of 5' upstream region, the 250-bp first exon, and 61 bp of the first intron was demonstrated to promote chloramphenicol acetyltransferase activity in a T-lymphoblast cell line and to have > 6-fold greater activity in a Jurkat T lymphoblast than in a Raji B-lymphoblast cell line. Our data suggest that these 5' sequences may contain elements that are important for the tissue-specific differences in deoxycytidine kinase expression. PMID- 8421672 TI - Phylogenic and ontogenic expression of hepatocellular bile acid transport. AB - The phylogenic and ontogenic expression of mRNA for the Na+/bile acid cotransporter was determined by Northern analysis utilizing a full-length cDNA probe recently cloned from rat liver. mRNA was detected in several mammalian species, including rat, mouse, and man, but could not be found in livers from nonmammalian species, including chicken, turtle, frog, and small skate. When expression of the bile acid transporter in developing rat liver was studied, mRNA was detected between 18 and 21 days of gestation, at the time when Na(+) dependent bile acid transport is first detected. Two hepatoma cell lines (HTC and HepG2), the latter of which is known to have lost the Na+/bile acid cotransport system, also did not express mRNA for this transporter. Finally, when mRNA from the lower vertebrate (the small skate) was injected into Xenopus oocytes, only a sodium-independent, chloride-dependent transport system for bile acids was expressed, confirming the integrity of the mRNA and consistent with prior functional studies of bile acid transport in this species. These findings establish that the Na+/bile acid cotransport mRNA is first transcribed in mammalian species, a process that is recapitulated late during mammalian fetal development in rat liver, and that this mRNA is lost in dedifferentiated hepatocytes. In contrast, the mRNA for a multispecific Na+/independent organic anion transport system is transcribed earlier in vertebrate evolution. PMID- 8421673 TI - Protein folding--what's the question? AB - The folding reactions of many small, globular proteins exhibit two-state kinetics, in which the folded and unfolded states interconvert readily without observable intermediates. Typically, the free energy difference, delta G, between the native and denatured states of such a protein is quite small, lying in the range of approximately -5 to -15 kcal/mol. We point out that, under these circumstances, a population of native-like molecules will persist, even in the presence of mutations sufficiently destabilizing to change the sign of delta G. Therefore, it is not energy per se that determines conformation. A corollary to this argument is that specificity--not stability--would be the more informative focus in future folding studies. PMID- 8421674 TI - The conserved lysine of the catalytic domain of protein kinases is actively involved in the phosphotransfer reaction and not required for anchoring ATP. AB - The study of the various protein kinases reveals that, despite their considerably diversity, they have evolved from a common origin. Eleven conserved subdomains have been described that encompass the catalytic core of these enzymes. One of these conserved regions, subdomain II, contains an invariant lysine residue present in all known protein kinase catalytic domains. Two facts have suggested that this conserved lysine of subdomain II is essential for binding ATP: (i) several investigators have demonstrated that this residue is physically proximal to the ATP molecule, and (ii) conservative substitutions at this site render the kinase inactive. However, these results are also consistent with a functional role of the conserved lysine of subdomain II in orienting or facilitating the transfer of phosphate. To study in more detail the role of subdomain II, we have generated mutants of the protein-tyrosine kinase pp56lck that have single amino acid substitutions within the area surrounding the conserved residue Lys-273 in subdomain II. When compared with wild-type pp56lck, these mutants displayed profound reductions in their phosphotransfer efficiencies and small differences in their affinities for ATP. Further, the substitution of arginine for Lys-273 resulted in a mutant protein unable to transfer the gamma-phosphate of ATP but able to bind 8-azido-ATP with an efficiency similar to that of wild-type pp56lck. These results suggest that the region including Lys-273 of subdomain II is involved in the enzymatic process of phosphate transfer, rather than in anchoring ATP. PMID- 8421675 TI - Phospholipid transmembrane domains and lateral diffusion in fibroblasts. AB - The lateral diffusion of fluorescent phospholipids in cultured Chinese hamster lung fibroblasts was examined by modulated fringe pattern photobleaching. When cells were labeled and maintained at 7 degrees C, the fluorescence remained localized at the plasma membrane. N-[6-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl amino)caproyl] sphingosylphosphocholine (C6-NBD-SphPCho) and 1-acyl-2-[6-(7 nitrobenz-2-oxa-1,3-diazol-4-yl-amino)caproyl] phosphatidylcholine (C6-NBD PtdCho) both diffused with the same apparent lateral diffusion coefficient (D1 approximately 0.3 x 10(-9) cm2/s). By contrast, the phosphatidylserine derivative (1-acyl-2-[6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl-amino)caproyl] phosphatidylserine (C6-NBD-Ptd-Ser)) gave rise to two diffusional components: a slow component, D1, analogous to that measured with the choline-containing lipids, and a fast component (D2 approximately 2 x 10(-9) cm2/s). The fast component only exists in ATP-containing cells. It was shown to be associated with C6-NBD-PtdSer translocated to the inner leaflet. This indicates that the two leaflets form very different membranous domains. At higher temperature, the same difference in mobility was observed between the choline-containing lipids and the aminolipid. However, with C6-NBD-SphPCho, a fraction of very slowly diffusing or quasi immobilized probes gradually appeared with time. This could be attributed to sphingomyelin located in small organelles after internalization. From the amplitude of this component registered at different intervals, we calculated that approximately 50% of the plasma membrane sphingomyelin is recycled in less than 30 min in Chinese hamster fibroblasts by an ATP- and microtubule-dependent process. PMID- 8421676 TI - A dominant truncation allele identifies a gene, STE20, that encodes a putative protein kinase necessary for mating in Saccharomyces cerevisiae. AB - This work reports the identification, characterization, and nucleotide sequence of STE20, a newly discovered gene involved in the Saccharomyces cerevisiae mating response pathway, to date one of the best understood signal transduction pathways. STE20 encodes a putative serine/threonine-specific protein kinase with a predicted molecular mass of 102 kDa. Its expression pattern is similar to that of several other protein kinases in the mating response pathway. Deletion of the kinase domain of STE20 causes sterility in both haploid mating types. This sterility can be partially suppressed by high-level production of STE12 but is not suppressible by high levels of STE4 or a dominant STE11 truncation allele. A truncation allele of STE20 was isolated that can activate the mating response pathway in the absence of exogenous mating pheromone. This allele causes dominant growth arrest that cannot be suppressed by deletions of STE4, STE5, STE7, STE11, or STE12. The allele is able to suppress the mating defect of a strain in which the STE20 kinase domain has been deleted, but not the mating defects of strains carrying mutations in STE4, STE5, STE7, STE11, or STE12. PMID- 8421677 TI - Mouse placental macrophages have a decreased ability to present antigen. AB - Large numbers of macrophages can be found in an animal's uteroplacental unit. This high concentration of macrophages suggests they must play an important role during placental development. To gain a better understanding of the functional capacity of placental macrophages, we have obtained a highly enriched placental macrophage culture and have derived several cell lines from this population. Both placental macrophages and cell lines show colony-stimulating factor 1-dependent growth, express Fc receptors, and can perform Fc-receptor-mediated phagocytosis. In addition, they express macrophage markers Mac-1, F4/80, and CD14. Although placental macrophages express major histocompatibility complex class II molecules constitutively, they display a decreased ability to present protein antigens to T cells. Since primary fetal liver macrophages of the same gestational stage also show a decreased ability to present antigens, this phenomenon may reflect a developmental stage of macrophages. PMID- 8421678 TI - Induction of protein-tyrosine-phosphatase activity by interleukin 6 in M1 myeloblastic cells and analysis of possible counteractions by the BCR-ABL oncogene. AB - Interleukin 6 (IL-6) induces in M1 myeloblastic cells growth arrest and terminal differentiation toward monocytes. It is reported here that IL-6 reduced by 5- to 20-fold the tyrosine phosphorylation of cellular proteins in these cells. The same-fold reduction was also observed in M1 cells that were transfected with the BCR-ABL deregulated protein kinase. In these stable clones, the levels of tyrosine phosphorylation of cellular proteins were 30- to 100-fold higher than in the parental cells. IL-6 did not reduce the expression levels or the inherent tyrosine kinase activity of BCR-ABL p210. By measuring the protein-tyrosine phosphatase (PTPase; protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48) activity in crude cell lysates, we found that protein dephosphorylation resulted, at least partially, from induction of PTPase activity by IL-6. The induction of PTPase in the BCR-ABL-transfected clones was not sufficient to confer the minimal protein phosphorylation levels characteristic of IL-6-treated cells. Yet, the transfected M1 clones showed normal growth and differentiation responses to IL-6. None of the gene responses to IL-6 including suppression in the levels of c-myc, c-myb, and cyclin A mRNA; junB and c-jun mRNA induction; and dephosphorylation of retinoblastoma protein were rescued by the BCR-ABL oncogene. The functional relevance of PTPase induction by IL-6 is discussed. PMID- 8421679 TI - Evidence to implicate translation by ribosomes in the mechanism by which nonsense codons reduce the nuclear level of human triosephosphate isomerase mRNA. AB - The abundance of the mRNA for human triosephosphate isomerase (TPI) is decreased to 20-30% of normal by frameshift and nonsense mutations that prematurely terminate translation within the first three-quarters of the reading frame. The decrease has been shown to be attributable to a reduced level of TPI mRNA that copurifies with nuclei. Given that the translational reading frame of an mRNA is assessed in the cytoplasm during protein synthesis, cytoplasmic and nuclear RNA processes may be linked. Alternatively, a nuclear mechanism may exist whereby in frame nonsense codons can be identified. To differentiate between these two possibilities, two distinct modulators of protein synthesis have been tested for the ability to influence the nonsense-codon-mediated reduction in the mRNA level. (i) A suppressor tRNA, which acts in trans to suppress an amber nonsense codon within TPI mRNA, and (ii) a hairpin structure in the 5' untranslated region of TPI mRNA, which acts exclusively in cis to inhibit initiation of TPI mRNA translation, were found, individually, and to a greater extent, together, to abrogate the decrease in mRNA. These results show that tRNA and ribosomes coordinately mediate the effect of a nonsense codon on the level of newly synthesized TPI mRNA. We suggest that the premature termination of TPI mRNA translation in the cytoplasm can reduce the level of TPI mRNA that fractionates with nuclei. PMID- 8421680 TI - A localized differentiation-inducing-factor sink in the front of the Dictyostelium slug. AB - Differentiation-inducing factor 1 [DIF-1; 1-(3,5-dichloro-2,6-dihydroxy-4 methoxyphenyl)-hexan-1-one] induces stalk cell differentiation during Dictyostelium development. It is present as a gradient in the multicellular slug, its lowest concentration being in the anterior. Here we demonstrate the existence of a localized sink for DIF-1, also in the anterior of the slug, which could be responsible for generating the DIF-1 gradient. DIF-1 is metabolized extensively by developing cells, initially by a mono-dechlorination. We used an enzyme assay for DIF-1 dechlorinase to examine its distribution in the slug. DIF-1 dechlorinase activity is 30-fold higher in prestalk cells (largely anterior) compared with prespore cells (posterior) when these are separated from each other on Percoll density gradients. Dissection experiments showed that DIF-1 dechlorinase is 25-fold enriched in the anterior 13% of the slug compared with the rest. These experiments also showed that DIF-1 dechlorinase is more anterior enriched than the standard prestalk markers, the ecmA and ecmB mRNAs. When cut from a slug, both prestalk and prespore fragments regulate to restore the missing cell type. Prespore fragments rapidly regain (by 30 min) a DIF-1 sink in their anteriors, and prestalk fragments restore a posterior zone with low DIF-1 dechlorinase by 4 hr after cutting. The reappearance of the DIF-1 sink in the anterior of prespore fragments is accomplished without detectable cell sorting and may, therefore, be in response to positional signals. Finally, a localized sink may provide a general way of producing a gradient of a signal substance in a developing embryo. PMID- 8421681 TI - In vivo analysis of Chlamydomonas chloroplast petD gene expression using stable transformation of beta-glucuronidase translational fusions. AB - We have used the Escherichia coli beta-glucuronidase (uidA) gene as a reporter gene to localize the promoter and analyze the function of the 5' untranslated region (UTR) of the Chlamydomonas chloroplast petD gene. Using particle bombardment, petD-uidA transcriptional and translational fusion genes were introduced into the chloroplast genome in the large inverted repeat flanking the atpB gene. In transformants carrying a petD-uidA transcriptional fusion, uidA mRNA accumulated but was not translated. However, in a translational fusion that included the entire petD 5' UTR, uidA mRNA accumulated and a high level of beta glucuronidase activity was detected. When approximately 70% of the petD 5' UTR was deleted from the translational fusion, uidA mRNA accumulation and beta glucuronidase activity decreased 4- to 6-fold and 8-fold, respectively. Run-on transcription assays demonstrated that all strains transcribe the uidA gene at equivalent rates. Our results show that sequences essential for translation reside in the petD 5' UTR and also that sequences within the 5' UTR directly or indirectly affect mRNA stability. The expression of beta-glucuronidase under the control of chloroplast transcriptional and translational signals will facilitate further studies of chloroplast gene regulatory mechanisms. PMID- 8421682 TI - Carboxyethyllysine in a protein: native carbonyl reductase/NADP(+)-dependent prostaglandin dehydrogenase. AB - Two different forms of the monomeric NADP(+)-linked prostaglandin dehydrogenase/carbonyl reductase were purified from human placenta and shown to differ by the modification of a lysine residue. The modified and the unmodified proteins were reproducibly recovered in a ratio of approximately 1:3, and both were chemically stable. The modified form was more acidic (pI approximately 7.4 versus pI approximately 7.7) but indistinguishable from the unmodified form in specificity and activity. Amino acid analysis, sequence analysis, mass spectrometry, and chemical synthesis identified the modified residue as N6-(1 carboxyethyl)lysine with C-2 of propionic acid attached to the side-chain N of Lys-238. This compound can be formed from the lysine residue and pyruvate via a Schiff base and subsequent reduction. The enzyme and its NAD(+)-dependent counterpart are distantly related (23% residue identity) and have the same family assignment to short-chain dehydrogenases. Alignments and model-building into the tertiary structure of 3 alpha/20 beta-hydroxysteroid dehydrogenase show that carbonyl reductase has an extra loop (positions 149-189) that forms a separate extension and replaces a backbone C-terminal beta-strand. This change affects the substrate pocket, explaining the different substrate specificities but conserves residues of known functional importance. Carboxyethyllysine at position 238 corresponds to a proteolysis-sensitive position in several short-chain dehydrogenases, less well-defined in the model but close to a surface, and is compatible with the accessibility and enzyme properties observed. PMID- 8421683 TI - Insulin down-regulates expression of the insulin-responsive glucose transporter (GLUT4) gene: effects on transcription and mRNA turnover. AB - Insulin rapidly represses expression of the gene encoding the insulin-responsive glucose transporter (GLUT4) in 3T3-L1 mouse adipocytes. Upon exposure to the hormone the cellular level of GLUT4 mRNA falls (t1/2 approximately 2.5 hr) to 20 30% of its initial level within 10 hr. This is followed by a similar decrease in the level of GLUT4 protein. Down-regulation of GLUT4 mRNA is a result of both rapid repression of transcription of the GLUT4 gene and an increased rate of turnover of the GLUT4 message. As a consequence of prolonged exposure to insulin, 3T3-L1 adipocytes lose their capacity for acute stimulation of hexose uptake by insulin. These findings provide an explanation for the resistance of glucose uptake to insulin in adipose tissue observed in non-insulin-dependent (type 2) diabetes mellitus, particularly that associated with hyperinsulinemia and obesity. PMID- 8421684 TI - Identification of an RNA binding site for human thymidylate synthase. AB - Previous studies from this laboratory have shown that human TS mRNA translation is regulated by its protein product in a negative autoregulatory manner. In this paper, we identify an RNA binding site for TS protein located within the first 188 nt of TS RNA. A 36-nt RNA sequence contained within this 188-nt fragment, corresponding to nt 75-110 and including the translational initiation site, binds TS protein with an affinity similar to that of both the full-length and the 188 nt TS RNA sequences. Variant RNAs with either a deletion or a mutation at the translational initiation region are unable to compete for TS protein binding. UV crosslinking studies reveal that an RNA fragment of approximately 36 nt is protected from RNase T1 digestion by TS protein binding. A second TS protein binding site is localized within the protein-coding region corresponding to nt 434-634. These findings demonstrate a specific interaction between human TS protein and its TS RNA and identify an RNA binding site that includes the translational initiation site. PMID- 8421685 TI - Differential expression of the two nonallelic proinsulin genes in the developing mouse embryo. AB - In the mouse, insulin is produced from two similar but nonallelic genes that encode proinsulins I and II. We have investigated expression of these two genes during mouse embryonic development, using a PCR to detect the two gene transcripts and immunocytochemistry to visualize the two corresponding proteins. At appearance of the dorsal pancreatic anlage at day 9.5 of gestation, both mRNAs could be detected in the embryos, and both proteins were present together in the same cells of the developing pancreas. At days 9.5 and 10.5, when the ventral anlage appears, there were fewer proinsulin II mRNAs than proinsulin I mRNAs. At day 12.5 this ratio was reversed. Proinsulin II mRNA, but not proinsulin I mRNA, could be detected at day 8.5 in the prepancreatic embryo. Proinsulin II mRNA, but not proinsulin I mRNA, was also found in the heads of embryos at day 9.5 and at all later stages studied. These results indicate that the two proinsulin genes are regulated independently, at least in part. They also suggest that insulin might play a role as a growth factor in the developing mouse brain. PMID- 8421686 TI - A mutation within intron 3 of the Pax-3 gene produces aberrantly spliced mRNA transcripts in the splotch (Sp) mouse mutant. AB - The splotch (Sp) mouse mutant displays defects in neural tube closure in the form of exencephaly and spina bifida. Recently, mutations in the Pax-3 gene have been described in the radiation-induced Spr and Sp2H alleles. This led us to examine the integrity of the Pax-3 gene and its cellular mRNA transcript in the original, spontaneously arising Sp allele. A complex mutation in the Pax-3 gene including an A-->T transversion at the invariant 3' AG splice acceptor of intron 3 was identified in the Sp/Sp mutant. This genomic mutation abrogates the normal splicing of intron 3, resulting in the generation of four aberrantly spliced mRNA transcripts. Two of these Pax-3 transcripts make use of cryptic 3' splice sites within the downstream exon, generating small deletions which disrupt the reading frame of the transcripts. A third aberrant splicing event results in the deletion of exon 4, while a fourth retains intron 3. These aberrantly spliced mRNA transcripts are not expected to result in functional Pax-3 proteins and are thus responsible for the phenotype observed in the Sp mouse mutant. PMID- 8421687 TI - Conserved nucleotide sequences in the open reading frame and 3' untranslated region of selenoprotein P mRNA. AB - Rat liver selenoprotein P contains 10 selenocysteine residues in its primary structure (deduced). It is the only selenoprotein characterized to date that has more than one selenocysteine residue. Selenoprotein P cDNA has been cloned from human liver and heart cDNA libraries and sequenced. The open reading frames are identical and contain a signal peptide, indicating that the protein is secreted by both organs and is therefore not exclusively produced in the liver. Ten selenocysteine residues (deduced) are present. Comparison of the open reading frame of the human cDNA with the rat cDNA reveals a 69% identity of the nucleotide sequence and 72% identity of the deduced amino acid sequence. Two regions in the 3' untranslated portion have high conservation between human and rat. Each of these regions contains a predicted stable stem-loop structure similar to the single stem-loop structures reported in 3' untranslated regions of type I iodothyronine 5'-deiodinase and glutathione peroxidase. The stem-loop structure of type I iodothyronine 5'-deiodinase has been shown to be necessary for incorporation of the selenocysteine residue at the UGA codon. Because only two stem-loop structures are present in the 3' untranslated region of selenoprotein P mRNA, it can be concluded that a separate stem-loop structure is not required for each selenocysteine residue. PMID- 8421688 TI - A cyclophilin-related protein involved in the function of natural killer cells. AB - Natural killer cells are non-major histocompatibility complex-restricted large granular lymphocytes that can recognize and destroy tumor cells without prior stimulation. A 150-kDa molecule on the surface of human natural killer cells was identified as a component of a putative tumor-recognition complex. We report here the isolation of cDNAs coding for the 150-kDa tumor-recognition molecule from human and mouse cDNA libraries. The amino terminus of the predicted protein contains a large hydrophobic region followed by a domain that is highly homologous to cyclophilin/peptidylprolyl cis-trans isomerase. The remainder of the protein is extremely hydrophilic and contains three homologous positively charged clusters. There are also three regions that contain extensive arginine- and serine-rich repeats. Comparison of the human and mouse predicted amino acid sequences revealed > 80% homology. PMID- 8421690 TI - Erosion kinetics of hydrolytically degradable polymers. AB - Degradable polymers are beginning to play an increasing role as materials for environmental and medical applications. Understanding factors that control erosion, such as bond cleavage and the dissolution and diffusion of degradation products, will be critical to the future development of these materials. Erosion kinetics, photomicroscopy, and infrared spectroscopy were used to understand the erosion mechanism of two families of degradable polymers, polyanhydrides and polyesters. Polyanhydrides exhibit behavior more characteristic of surface erosion, whereas the polyesters exhibit bulk erosion patterns. Control of erosion times from a few days to several years can be achieved by a judicious choice of monomer units and bond selection. PMID- 8421689 TI - A recombinant immunotoxin that is active on prostate cancer cells and that is composed of the Fv region of monoclonal antibody PR1 and a truncated form of Pseudomonas exotoxin. AB - Monoclonal antibody PR1 binds to the surface of normal prostate cells and to adenocarcinomas of the prostate. The cDNAs coding for the heavy and light chain variable regions of monoclonal antibody PR1 were cloned by PCR techniques. A recombinant toxin was then constructed that has the heavy chain variable region of monoclonal antibody PR1 connected to the light chain variable region by a flexible peptide linker to create a single-chain Fv; the Fv in turn is fused to a truncated form of Pseudomonas exotoxin. The resulting recombinant immunotoxin PR1(Fv)-PE38KDEL was produced in Escherichia coli and accumulated in inclusion bodies. After denaturation and renaturation, active monomeric molecules with a molecular mass of approximately 65 kDa were purified to homogeneity. PR1(Fv) PE38KDEL binds specifically to cells containing the PR1 antigen and is very cytotoxic toward a subset of LNCaP cells that express the PR1 antigen on their surface. PMID- 8421691 TI - Barium-induced exocytosis is due to internal calcium release and block of calcium efflux. AB - The concentration of cytosolic free Ca2+ ([Ca2+]i) and the release of tritiated norepinephrine ([3H]NE) were monitored during Ba2+ stimulation of sympathetic neurons cultured from chick embryos. Ba2+ (2.5 mM in Ca(2+)-free medium) caused a rise in [Ca2+]i in all regions (cell bodies, neurites, and growth cones) of sympathetic neurons and evoked [3H]NE release in the absence of other stimuli. The increase in [Ca2+]i and release of [3H]NE were sustained for up to 30 min in the presence of Ba2+. When Ba(2+)-stimulated cells were immediately washed in Ca(2+)-free Ba(2+)-free EGTA solution, both the elevated [Ca2+]i and [3H]NE release returned to basal levels, with similar, fast, time courses. Ba2+ also blocked Ca2+ efflux from neurons loaded with 45Ca. We conclude from the parallel effects of Ba2+ on [Ca2+]i and [3H]NE release that Ba2+ stimulates exocytosis by a Ca(2+)-dependent mechanism. The Ba(2+)-induced rise in [Ca2+]i is a result of two separate actions: (i) the release of Ca2+ from intracellular sites and (ii) an effective block of Ca2+ extrusion. The ability of Ba2+ to release Ca2+ in growth cones that are insensitive to caffeine suggests that Ba2+ may displace Ca2+ from binding sites other than endoplasmic reticulum. PMID- 8421692 TI - A possible glycine radical in anaerobic ribonucleotide reductase from Escherichia coli: nucleotide sequence of the cloned nrdD gene. AB - During anaerobic growth of Escherichia coli an oxygen-sensitive ribonucleoside triphosphate reductase, different from the aerobic ribonucleoside diphosphate reductase (EC 1.17.4.1), produces the deoxyribonucleoside triphosphates required for DNA replication. The gene for the anaerobic enzyme has now been cloned and was found to contain a 2136-nucleotide coding region, corresponding to 712 amino acid residues, and an Fnr binding site 228 base pairs upstream of the initiator ATG. The deduced amino acid sequence shows 72% identity to a gene of coliphage T4, sunY, hitherto of unknown function, suggesting that the virus codes for its own anaerobic reductase. The location of an organic free radical formed during activation of the bacterial anaerobic reductase is proposed to be on Gly-681, since the pentapeptide RVCGY at positions 678-682 shows a striking similarity to the C-terminal sequence. RVSGY, of pyruvate formate-lyase. During activation of the anaerobically induced pyruvate formate-lyase, the glycine residue of the pentapeptide becomes an organic radical [Wagner, A. F. V., Frey, M., Neugebauer, F. A., Schafer, W. & Knappe, J. (1992) Proc. Natl. Acad. Sci. USA 89, 996-1000]. The gene for the anaerobic reductase is located at a position around 96 min on the E. coli genomic map. PMID- 8421693 TI - Paradoxical structure and function in a mutant human insulin associated with diabetes mellitus. AB - The solution structure of a diabetes-associated mutant human insulin (insulin Los Angeles; PheB24-->Ser) was determined by 13C-edited NMR spectroscopy and distance geometry/simulated annealing calculations. Among vertebrate insulins PheB24 is invariant, and in crystal structures the aromatic ring appears to anchor the putative receptor-binding surface through long-range packing interactions in the hydrophobic core. B24 substitutions are of particular interest in relation to the mechanism of receptor binding. In one analogue ([GlyB24]insulin), partial unfolding of the B chain has been observed with paradoxical retention of near native bioactivity. The present study of [SerB24]insulin extends this observation: relative to [GlyB24]insulin, near-native structure is restored despite significant loss of function. To our knowledge, our results provide the first structural study of a diabetes-associated mutant insulin and support the hypothesis that insulin undergoes a change in conformation on receptor binding. PMID- 8421694 TI - Genome mapping by nonrandom anchoring: a discrete theoretical analysis. AB - As part of our effort to construct a physical map of the genome of the fission yeast Schizosaccharomyces pombe, we have made theoretical predictions for the progress expected, as measured by the expected length fraction of island coverage and by the expected properties of the anchored islands such as the number and the size of islands. Our experimental strategy is to construct a random clone library and screen the library for clones having unique sequence at both ends. This scheme is essentially the same as the clone-limited double sequence-tagged-site selection scheme which was used in a computer simulation by Palazzolo et al. [Palazzolo, M. J., Sawyer, S. A., Martin, C. H., Smoller, D. A. & Hartl, D. L. (1991) Proc. Natl. Acad. Sci. USA 88, 8034-8038]. Both simulation and ongoing experiments in our laboratory have shown that the nonrandom anchoring method is far superior to random anchoring. In this paper, we propose a theoretical model to explain the simulated data and the experimental data. PMID- 8421695 TI - The trans-activator tax of human T-cell leukemia virus type 1 (HTLV-1) interacts with cAMP-responsive element (CRE) binding and CRE modulator proteins that bind to the 21-base-pair enhancer of HTLV-1. AB - The trans-activator protein Tax of human T-cell leukemia virus type 1 (HTLV-1) activates the viral 21-base-pair (bp) enhancer in the long terminal repeat and has been suggested to associate indirectly with the enhancer DNA. To demonstrate this, we used DNA-affinity precipitation assay and detected the Tax protein in 21 bp DNA-protein complexes isolated from HTLV-1-infected cells. To identify cellular components in the complexes, we tested various 21-bp DNA-binding proteins by gel electrophoretic mobility-shift assay. Each binding protein gave a shifted band of each 21-bp DNA-protein complex, and exogenously added Tax protein further shifted these bands of cAMP-responsive element (CRE) binding protein (CREB) and CRE modulator but did not shift other bands. Anti-Tax antibodies blocked formation of the complex, indicating complex formations of [Tax-CREB(or CRE modulator)-21-bp DNA]. The formations of these complexes paralleled the functional activities of Tax mutants. Furthermore, the Tax-CREB complex was detected in a nuclear extract of HTLV-1-infected cells, and the Tax-CREB-21-bp DNA complex was demonstrated as a major component of Tax complexes containing the 21-bp DNA probe. These observations indicate that Tax protein binds to CREB and CRE modulator and the complexes then bind to the 21-bp enhancer, suggesting that the complex binding to the enhancer mediates trans-activation of transcription. PMID- 8421696 TI - Expression of wild-type and mutant bovine pancreatic ribonuclease A in Escherichia coli. AB - Wild-type ribonuclease A and five mutants thereof have been expressed in Escherichia coli as fusion proteins by using a T7 expression system. The five mutants are C[65-72]S, C[40-95]S, C[58-110]S, C[26-84]S, and K41G. The expressed fusion protein formed inclusion bodies which were then cleaved by factor Xa. The cleaved ribonuclease A was isolated as unfolded (sulfonated), soluble protein which was subsequently folded. This expression system can be used to produce mutants of ribonuclease A in yields suitable for folding and structural studies. All four native three-disulfide mutants exhibited enzymatic activity (5-30%), although only two were thermally stable at room temperature, demonstrating that no single native disulfide bond is essential for folding. The K41G mutant was enzymatically inactive with cyclic cytidine monophosphate as substrate. PMID- 8421697 TI - Rhizobium NodB protein involved in nodulation signal synthesis is a chitooligosaccharide deacetylase. AB - The common nodulation genes nodABC are conserved in all rhizobia and are involved in synthesis of a lipooligosaccharide signal molecule. This bacterial signal consists of a chitooligosaccharide backbone, which carries at the nonreducing end a fatty acyl chain. The modified chitooligosaccharide molecule triggers development of nodules on the roots of the leguminous host plant. To elucidate the specific role of the NodB protein in nodulation factor synthesis, we have purified recombinant NodB and determined its biochemical role by direct assays. Our data show that the NodB protein of Rhizobium meliloti deacetylates the nonreducing N-acetylglucosamine residue of chitooligosaccharides. The monosaccharide N-acetylglucosamine is not deacetylated by NodB. In the pathway of Nod factor synthesis, deacetylation at the nonreducing end of the oligosaccharide backbone may be a necessary requirement for attachment of the fatty acyl chain. PMID- 8421698 TI - A histidine protein kinase is involved in polar organelle development in Caulobacter crescentus. AB - Mutations having pleiotropic effects on polar organelle development (pod) in Caulobacter crescentus have been identified and shown to occur in at least 13 genes scattered throughout the genome. Mutations at each locus affect a unique combination of polar traits, suggesting that complex interactions occur among these genes. The DNA sequence of one of these genes, pleC, indicates that it is homologous to members of the family of histidine protein kinase genes. Membes of this family include the senor components of the bacterial two-component regulatory systems. Furthermore, in vitro experiments demonstrated that the PleC protein was capable of autophosphorylation. These results suggest that the PleC protein (and perhaps the proteins encoded by the other pod genes as well) regulates the expression of genes involved in polar organelle development through the phosphorylation of key regulatory proteins. The use of a phosphorelay system cued to internal changes in the cell would provide a mechanism for coordinating major changes in gene expression with the completion of specific cell cycle events. PMID- 8421699 TI - Three small nucleolar RNAs of unique nucleotide sequences. AB - Three small RNA species were detected in human cells, and their cDNAs were synthesized and cloned. These RNAs are nucleolar, are 207, 154, and 135 nucleotides long, and are named E1, E2, and E3, respectively, and their unique nucleotide sequences suggest that they may belong to an additional family of small nucleolar RNAs. The 5' ends of these three RNAs do not appear to have a trimethylguanosine cap or another type of cap. Apparent homologs of these three RNAs were detected in mouse, rabbit, and frog cells, suggesting their universal importance. They are housekeeping RNA species, since they are present in all rabbit tissues analyzed. PMID- 8421700 TI - On the nature of the protein folding code. AB - This paper investigates quantitatively the characteristics of the local folding code. The overlapping four-residue fragments which make up the amino acid sequences of 114 proteins are divided into classes on the basis of the physical properties of their constituent amino acids. The distribution of structural types associated with each class of sequence fragment is determined and compared with an ensemble of random structural distributions of the same size selected from the actual protein structures. A criterion is proposed, based on the relative entropies of the two types of distribution, and on a hypothesis as to the characteristics of fragments which code for local structure, that makes it possible to identify those four-residue sequence elements which encode specific time-averaged structure. It is determined that, by this criterion, only 60-70% of the four-residue fragments encode specific structures. It is suggested that the remaining sequence fragments intrinsically encode susceptibility to conformational alteration under the influence of long-range interactions and that this susceptibility is required for correct folding of the molecule. This feature introduces an inherent indeterminacy into the local folding code. The implications of this observation for the prediction of protein structure by various methods are briefly discussed. PMID- 8421701 TI - Inhibitory effects of antisense oligodeoxynucleotides targeting c-myc mRNA on smooth muscle cell proliferation and migration. AB - Smooth muscle cell (SMC) proliferation and migration play pivotal roles in restenosis following angioplasty. c-myc is an immediate early response gene induced by various mitogens, and several lines of evidence derived from experiments using transformed or hematopoietic cell lines, or transgenic mice, suggest its protein product plays a role in numerous signaling transduction pathways, including those modulating cell division. We therefore reasoned that a strategy employing oligodeoxynucleotides (ODNs) complementary to c-myc mRNA (antisense ODNs) might be potent inhibitors of SMC proliferation and, perhaps, of SMC migration. To evaluate this concept, we tested several antisense ODNs targeted to c-myc mRNA (15- or 18-mer ODNs complementary to different c-myc mRNA sequences) by introducing them individually into the medium of cultured rat aortic SMCs. Phosphoroamidate-modified ODNs were employed to retard degradation. Antisense ODNs inhibited, in a concentration-dependent manner, SMC proliferation and SMC migration. Maximal inhibitory effect was 50% for proliferation and > 90% for migration. These effects were associated with decreased SMC expression of c myc-encoded protein by Western immunoblotting and immunocytochemical staining. ODNs with the same nucleotides but a scrambled sequence caused no effect. These results indicate that the c-myc gene product is involved in the signal transduction pathways mediating SMC proliferation and migration in the in vitro model we employed. The results also suggest a potential role of antisense strategies designed to inhibit c-myc expression for the prevention of coronary restenosis. PMID- 8421702 TI - Molecular evolution of the myosin family: relationships derived from comparisons of amino acid sequences. AB - To examine the evolutionary relationships between members of the myosin family, we have used two different phylogenetic methods, distance matrix and maximum parsimony, to analyze all available myosin head sequences. We find that there are at least three equally divergent classes of myosin, demonstrating that the current classification of myosin into only two classes needs to be reexamined. In the myosin II class, smooth muscle myosin is more closely related to nonmuscle myosin than to striated muscle myosin, implying that smooth muscle and skeletal muscle myosins were independently derived from nonmuscle myosin and suggesting that similarities between these types of muscle are the result of convergent evolution. The grouping of head sequences produced by phylogenetic analysis is consistent with classifications based on enzymology and structural localization and is generally consistent with grouping based on common tail structure elements. This result demonstrates that specific head sequences are tightly coupled to specific tail sequences throughout evolution and challenges the idea that myosin heads are freely interchangeable units whose unique function is determined only by the tail structure to which it is attached. PMID- 8421703 TI - Construction of chimeric alleles with altered specificity at the b incompatibility locus of Ustilago maydis. AB - Multiallelic incompatibility systems found in many fungi and plants function to limit inbreeding by mediating self versus nonself recognition. The plant pathogenic fungus Ustilago maydis has a locus called b that governs incompatibility. Two multiallelic genes, bE and bW, are present at the b locus. Fusion of haploid strains carrying different alleles at bE and bW establishes an infectious dikaryon capable of pathogenesis on maize (Zea mays). Cells carrying a single type of b locus, whether haploid or dikaryotic, are nonpathogenic. To identify sequences within the bE gene that determine allelic specificity, targeted gene replacement was employed to produce a series of chimeras between the b1E and b2E alleles. Incompatibility tests with strains carrying the chimeric alleles identified a 30- to 48-amino acid region responsible for specificity. Suprisingly, the chimeras with recombination points within this region had a specificity different from both parent alleles. Overall, these results define an important domain in bE involved in self versus nonself recognition. PMID- 8421704 TI - Developmental regulation of a murine T-cell-specific tyrosine kinase gene, Tsk. AB - Protein-tyrosine kinases have been implicated in signal transduction in T lymphocytes after stimulation of many cell-surface molecules, including the T cell antigen receptor, CD4, CD8, CD2, CD5, and CD28. Yet the identities of many of these tyrosine kinases remain unknown. We have isolated a murine tyrosine kinase gene, called Tsk for T-cell-specific kinase, that appears to be exclusively expressed in T lymphocytes. The Tsk cDNA clone encodes a polypeptide of 70 kDa, which is similar in sequence to both the src and abl families of tyrosine kinases. Sequence comparisons also indicate that Tsk contains one src homology region 2 domain and one src-homology 3 domain but lacks the negative regulatory tyrosine (src Tyr-527) common to src-family kinases. In addition, Tsk expression is developmentally regulated. Steady-state Tsk mRNA levels are 5- to 10-fold higher in thymocytes than in peripheral T cells and increase in the thymus during mouse development from neonate to adult. Furthermore, Tsk is expressed in day 14 fetal thymus, suggesting a role for Tsk in early T-lymphocyte differentiation. PMID- 8421705 TI - Pleiotrophin transforms NIH 3T3 cells and induces tumors in nude mice. AB - The pleiotrophin (PTN) gene (Ptn) encodes an 18-kDa protein that is highly conserved among mammalian species and that functions as a weak mitogen and promotes neurite-outgrowth activity in vitro. To further investigate the role PTN plays in regulating cell growth, we overexpressed the bovine PTN cDNA and now show that PTN phenotypically transforms NIH 3T3 cells, as evidenced by increased cell number at confluence, focus formation, anchorage-independent growth, and tumor formation in the nude mouse. The results demonstrate that the Ptn gene has the potential to regulate NIH 3T3 cell growth and suggest that PTN may influence abnormal cell growth in vivo. PMID- 8421706 TI - Protease inhibitors and indoleamines selectively inhibit cholinesterases in the histopathologic structures of Alzheimer disease. AB - Neurofibrillary tangles and amyloid plaques express acetylcholinesterase and butyrylcholinesterase activity in Alzheimer disease. We previously reported that traditional acetylcholinesterase inhibitors such as BW284C51, tacrine, and physostigmine were more potent inhibitors of the acetylcholinesterase in normal axons and cell bodies than of the acetylcholinesterase in plaques and tangles. We now report that the reverse pattern is seen with indoleamines (such as serotonin and its precursor 5-hydroxytryptophan), carboxypeptidase inhibitor, and the nonspecific protease inhibitor bacitracin. These substances are more potent inhibitors of the cholinesterases in plaques and tangles than of those in normal axons and cell bodies. These results show that the enzymatic properties of plaque and tangle-associated cholinesterases diverge from those of normal axons and cell bodies. The selective susceptibility to bacitracin and carboxypeptidase inhibitor indicates that the catalytic sites of plaque and tangle-bound cholinesterases are more closely associated with peptidase or protease-like properties than the catalytic sites of cholinesterases in normal axons and cell bodies. This shift in enzymatic affinity may lead to the abnormal protein processing that is thought to play a major role in the pathogenesis of Alzheimer disease. The availability of pharmacological and dietary means for altering brain indoleamines raises therapeutic possibilities for inhibiting the abnormal cholinesterase activity associated with Alzheimer disease. PMID- 8421707 TI - Expression of the human neuropeptide tyrosine Y1 receptor. AB - Neuropeptide tyrosine (NPY) is the predominant peptide in the innervation of many human tissues and is considered to play a role in the regulation of blood flow, gastrointestinal secretion and motility, and renal function. Three NPY receptors have been identified (Y1, Y2, and Y3) and the cDNAs encoding the human Y1 and bovine Y3 receptors have recently been cloned. We have demonstrated the expression of the Y1 receptor subtype in several fetal and adult human tissues, including the colon, kidney, adrenal gland, heart, and placenta. A single transcript was identified (approximately 2.2 kb) and localized in tissue sections by in situ hybridization. In the colon the receptor is expressed in the mucosa and basal glands, as well as the myenteric and submucous plexuses. Y1 receptor mRNA was detected in renal collecting ducts, loop of Henle, and juxtaglomerular apparatus and in the syncytiotrophoblast layer of placental villi. Fetal aorta and adult intramyocardial, colonic, and renal blood vessels also exhibited receptor expression, localized to the intima as well as the media. The distribution of Y1 receptor expression correlates with that of NPY-immunoreactive nerves and the apparent actions of NPY in the intestine, kidney, and heart. Although the placenta is devoid of nerves, an NPY-like transcript was detected in the villous trophoblast layer. The results indicate a tissue-specific regulation of NPY Y1 receptor expression. PMID- 8421708 TI - Acute hypoxia increases ornithine decarboxylase activity and polyamine concentrations in fetal rat brain. AB - The cellular responses to hypoxia are poorly understood. To test the hypothesis that ornithine decarboxylase (ODC; L-ornithine carboxy-lyase; EC 4.1.1.17) activity and polyamine concentrations change in response to acute hypoxia, we performed the following studies. Pregnant Sprague-Dawley rats inspired various O2 concentrations (9-21%) for various time periods (0.5-48 h) from days 15 to 21 of gestation. In fetal brains we measured the activity of ODC, ODC mRNA, and polyamines. In response to 4-h acute mild hypoxia, ODC activity in fetal rat brain (cerebrum, cerebellum, and hippocampus) increased to 330-450% from control values (P < 0.001), after which it declined to control levels in 6-8 h. The 4-h ODC response varied inversely with inspired O2 concentration and was not mimicked by beta 2 agonist or blocked by beta 2-antagonist administration. The ODC response was associated with an increase in fetal brain putrescine concentration to 190% above control at 4-6 h (P < 0.01) and an increase in the polyamines spermidine and spermine to about 115% above control at 6-8 h. We also observed that ODC mRNA increased significantly after 2-4 h of hypoxia. ODC activity and polyamine concentrations appear to be useful enzymatic markers for fetal brain hypoxia. The magnitude and time course of the acute hypoxic ODC increase were similar to responses to extracellular signals that result in differentiation or cell growth. Thus, the well-defined and regulated ODC activity response may represent a protective mechanism in brain to hypoxia. PMID- 8421709 TI - Regulation of cysteine-rich intestinal protein by dexamethasone in the neonatal rat. AB - The cysteine-rich intestinal protein (CRIP) is an intestinal zinc-binding protein containing a single copy of a cysteine-rich domain known as the LIM motif. CRIP mRNA and protein levels increased in the rat small intestine throughout the suckling period, reaching highest levels by the late weanling stage. A similar developmental pattern of CRIP protein levels was also detected by an increase in zinc binding to CRIP-containing HPLC fractions of intestinal cytosol. Administration of the synthetic glucocorticoid hormone dexamethasone to neonates caused the precocious rise of CRIP mRNA and protein. In adult rats, CRIP mRNA levels were not significantly altered by dexamethasone. Maximal CRIP mRNA content was detected in cells from the mid-villus, as confirmed by expression of cryptdin mRNA. In this report we show the glucocorticoid regulation of the LIM motif containing protein CRIP and suggest that glucocorticoid hormones play a role in developmental regulation of CRIP. PMID- 8421710 TI - Possible role of mammalian secretory group II phospholipase A2 in T-lymphocyte activation: implication in propagation of inflammatory reaction. AB - Both 2-lysophosphatidylcholine and cis-unsaturated fatty acids were previously shown to intensify agonist-induced cellular responses by enhancing the diacylglycerol-dependent activation of protein kinase C. Consistent with these observations, extracellular, secretory group II phospholipase A2, when added directly to human resting T lymphocytes, greatly potentiates their activation that was induced by a membrane-permeant diacylglycerol and ionomycin, as determined by the expression of the alpha subunit of the interleukin 2 receptor and thymidine incorporation into DNA. Diacylglycerol and ionomycin were essential for this cellular response, and phospholipase A2 alone showed no effect. The amount of 2-lysophosphatidylcholine produced in these cells by the exogenous phospholipase A2 was greatly increased in the presence of diacylglycerol and ionomycin, suggesting that the membrane phospholipids became susceptible to the phospholipase A2 when protein kinase C was activated. The results suggest that phospholipase A2 secreted into inflammatory sites plays a role in the propagation of cellular responses. Protein kinase C may function in the hydrolysis of membrane phospholipids by the exogenous phospholipase A2, and the products of this phospholipid hydrolysis enhance agonist-induced protein kinase C activation, thereby intensifying cellular responses. PMID- 8421711 TI - Specific activation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the gamma or zeta subunits of the immunoglobulin and T-cell receptors. AB - The generation of tumor-specific lymphocytes and their use in adoptive immunotherapy is limited to a few malignancies because most spontaneous tumors are very weak or not at all immunogenic. On the other hand, many anti-tumor antibodies have been described which bind tumor-associated antigens shared among tumors of the same histology. Combining the variable regions (Fv) of an antibody with the constant regions of the T-cell receptor (TCR) chains results in chimeric genes endowing T lymphocytes with antibody-type specificity, potentially allowing cellular adoptive immunotherapy against types of tumors not previously possible. To generalize and extend this approach to additional lymphocyte-activating molecules, we designed and constructed chimeric genes composed of a single-chain Fv domain (scFv) of an antibody linked with gamma or zeta chains, the common signal-transducing subunits of the immunoglobulin receptor and the TCR. Such chimeric genes containing the Fv region of an anti-trinitophenyl antibody could be expressed as functional surface receptors in a cytolytic T-cell hybridoma. They triggered interleukin 2 secretion upon encountering antigen and mediated non major-histocompatibility-complex-restricted hapten-specific target cell lysis. Such chimeric receptors can be exploited to provide T cells and other effector lymphocytes, such as natural killer cells, with antibody-type recognition directly coupled to cellular activation. PMID- 8421712 TI - Apolipoprotein J expression at fluid-tissue interfaces: potential role in barrier cytoprotection. AB - Apolipoprotein J (apoJ) is a sulfated secreted glycoprotein that exhibits ubiquitous expression, evolutionary conservation, and diverse tissue inducibility. It has been proposed to have roles in programmed cell death, sperm maturation, complement regulation, and lipid transport. To identify cell types that synthesize apoJ and to aid evaluation of its function, we screened mouse and human tissues by in situ hybridization. ApoJ was expressed at high levels in an array of specialized cell types of adult and fetal mouse tissues and in similar cell types of human tissues. Most of these cell types are highly secretory and form the cellular interfaces of many fluid compartments. This group includes epithelial boundary cells of the esophagus, biliary ducts, gallbladder, urinary bladder, ureter, kidney distal convoluted tubules, gastric glands, Brunner's glands, choroid plexus, ependyma, ocular ciliary body, endometrium, cervix, vagina, testis, epididymus, and visceral yolk sac. Several nonepithelial secretory cell types that express high levels of apoJ also line fluid compartments, such as synovial lining cells and ovarian granulosa cells. In the context of its known biochemical properties, this expression pattern suggests that localized synthesis of apoJ serves to protect a variety of secretory, mucosal, and other barrier cells from surface-active components of the extracellular environment. PMID- 8421713 TI - Abnormal activation of H+ conductance in NADPH oxidase-defective neutrophils. AB - To combat bacterial infection, phagocytes generate superoxide (O2-) and other microbicidal oxygen radicals. NADPH oxidase, the enzyme responsible for O2- synthesis, is deficient in chronic granulomatous disease (CGD) patients. Although O2- generation is accompanied by a large burst of metabolic acid production, intracellular pH (pHi) remains near neutrality due to the concomitant stimulation of H+ extrusion. Three major pathways contribute to pHi regulation in activated phagocytes: Na+/H+ exchange, vacuolar-type H+ pumps, and a H+ conductance. The present study analyzed the relationship between activation of the NADPH oxidase and stimulation of the H+ extrusion mechanisms in human blood neutrophils. Phorbol ester-induced activation of Na+/H+ exchange and H+ pumping occurred normally in cells from CGD patients. Unlike normal individuals, however, CGD patients were unable to activate the H+ conductive pathway. Thus, activation of the H+ conductance appears to be contingent on the assembly of a functional NADPH oxidase. These findings imply a dual role of the NADPH oxidase in O2- synthesis and in the regulation of pHi. The oxidase (or some components thereof) may itself undertake H+ translocation or, alternatively, may signal the activation of a separate H+ conducting entity. PMID- 8421714 TI - Transforming growth factor beta 1 null mutation in mice causes excessive inflammatory response and early death. AB - To delineate specific developmental roles of transforming growth factor beta 1 (TGF-beta 1) we have disrupted its cognate gene in mouse embryonic stem cells by homologous recombination to generate TGF-beta 1 null mice. These mice do not produce detectable amounts of either TGF-beta 1 RNA or protein. After normal growth for the first 2 weeks they develop a rapid wasting syndrome and die by 3-4 weeks of age. Pathological examination revealed an excessive inflammatory response with massive infiltration of lymphocytes and macrophages in many organs, but primarily in heart and lungs. Many lesions resembled those found in autoimmune disorders, graft-vs.-host disease, or certain viral diseases. This phenotype suggests a prominent role for TGF-beta 1 in homeostatic regulation of immune cell proliferation and extravasation into tissues. PMID- 8421716 TI - Measurement validity in physical therapy research. AB - This article considers the role of measurement validity within physical therapy research. The concept of measurement validity is identified as a component of internal validity, and it is differentiated from the notion of reliability; these concepts are related to systematic and random sources of error, respectively. Using examples from physical therapy and rehabilitation, four main types of validity are reviewed: face validity, criterion-related validity, content validity, and construct validity. The differing implications of these types of validity for quantitative and qualitative research are discussed. Three principal areas of concern are then addressed, based on a critical discussion of selected examples from the literature. First, it is argued that validity is often poorly distinguished from the allied concept of reliability and that purported claims for validity often only demonstrate reliability. Second, it is claimed that validity is too often neglected in favor of reliability, and specific examples relating to gait analysis are put forward to support this argument. Third, some of the methodological difficulties that may occur when attempts are made to demonstrate validity are considered. The article concludes with a plea for a closer focus on the issue of measurement validity within physical therapy research. PMID- 8421715 TI - Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African and American visceral leishmaniasis. AB - We report the cloning of a Leishmania chagasi antigen gene and an evaluation of leishmaniasis patient antibody responses to the recombinant protein, rK39. rK39 contains a 39-amino acid repeat that is part of a 230-kDa protein predominant in L. chagasi tissue amastigotes. Sequence analyses showed this protein, LcKin, to be related to the kinesin superfamily of motor proteins. Southern blot analyses demonstrated LcKin-related sequences in seven species of Leishmania, with conservation of the repeat between L. chagasi and Leishmania donovani. Serological evaluation revealed that 98% (56 of 57) of Brazilian and 100% (52 of 52) of Sudanese visceral leishmaniasis patients have high antibody levels to the rK39 repeat. Detectable anti-K39 antibody was virtually absent in cutaneous and mucosal leishmaniasis patients and in individuals infected with Trypanosoma cruzi. The data show that rK39 may replace crude parasite antigens as a basis for serological diagnosis of visceral leishmaniasis. PMID- 8421717 TI - A comparison of photographic and transparency-based methods for measuring wound surface area. AB - BACKGROUND AND PURPOSE: The purposes of this study were to compare test-retest reliability of measurements obtained by the use of a photographic method and those obtained by the use of a transparency method and to compare wound surface area (WSA) measurements obtained by these two methods. SUBJECTS: Twenty inpatients (18 male, 2 female), aged 31 +/- 16 years (mean +/- SD), participated in the study. METHODS: Tracings of ulcer borders generated by the photographic and transparency methods were digitized to obtain WSA measurements. To assess intrarater reliability for each method, 5 ulcers were measured on two occasions. The magnitude of WSA measurements obtained by the photographic and transparency methods was compared in 22 ulcers measured on one occasion and in 16 ulcers measured at 5-day intervals for 20 days. RESULTS: Intraclass correlation coefficients (ICCs) were high (ICC = .99) for each method, indicating reliability of measurements. The WSA measurements did not differ between photographic and transparency methods, either at one occasion or over a 20-day period. CONCLUSION AND DISCUSSION: We concluded that the photographic and transparency methods, as applied in this study of ulcers, provided equivalently reliable measurements and that WSA measurements obtained by the two methods were equivalent. The transparency method was more economical than the photographic method in terms of time and equipment requirements. PMID- 8421718 TI - Goal attainment scaling. PMID- 8421719 TI - Evaluation of soft foot orthotics in the treatment of patellofemoral pain syndrome. AB - BACKGROUND AND PURPOSE: The effectiveness of soft foot orthotics in the treatment of patients who have patellofemoral pain syndrome was investigated. SUBJECTS: Subjects were 20 adolescent female patients, aged 13 to 17 years (mean = 14.8, SD = 1.2), who were diagnosed with patellofemoral pain syndrome and who exhibited excessive forefoot varus or calcaneal valgus. METHODS: Subjects were randomly assigned to one of two groups: a control group (n = 10), which took part in an exercise program, or a treatment group (n = 10), which used soft foot orthotics in addition to participating in the exercise program. The exercise program consisted of quadriceps femoris and hamstring muscle strengthening and stretching exercises. A visual analogue scale was used to assess the level of pain of the subjects over an 8-week period. RESULTS: Both the treatment and control groups demonstrated a significant decrease in the level of pain, but the improvement of the treatment group was significantly greater than that of the control group. CONCLUSION AND DISCUSSION: The results suggest that in addition to an exercise program, the use of soft foot orthotics is an effective means of treatment for the patient with patellofemoral pain syndrome. PMID- 8421720 TI - Reliability of the scores for the finger-to-nose test in adults with traumatic brain injury. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine the intrarater and interrater reliability of measurements of three clinical features of coordination based on the performance of the "finger-to-nose" test. SUBJECTS: Thirty-seven persons with traumatic brain injury (26 male, 11 female), aged 17 to 64 years (mean = 29.1, SD = 9.9), participated in the study. METHODS: Each subject's performance was videotaped and evaluated for the right and left upper extremities (UEs) (two trials each) with respect to the following variables: time of execution, degree of dysmetria, and degree of tremor (four-point ordinal ratings). One year later, five experienced physical therapists (including the original investigator) independently rated each patient's videotaped performance in the same manner as described above. RESULTS: Intraclass correlation coefficients (ICC[3,1]) for intrarater reliability were .971 and .986 and ICCs for interrater reliability were .920 and .913 for right and left UEs, respectively, for the time of execution. A generalized Kappa statistic of .54 was calculated for the scoring of dysmetria (both UEs), and Kappa statistics calculated for the scoring of tremor were .18 and .31 for right and left UEs, respectively. Interrater reliability was lower for the scoring of these variables and varied from .36 to .40 for dysmetria and from .27 to .26 for tremor (right and left UEs, respectively). CONCLUSION AND DISCUSSION: These results indicate that physical therapists demonstrate low reliability in assessment of the presence of dysmetria and tremor using videotaped performances of the finger-to nose test. The results suggest, however, that therapists reliably measure the time of execution of this test. If the limitations associated with therapists' capacity for objective measurement of subjective phenomena cannot be overcome (eg, by establishment of more definitive scoring criteria for the measures of dysmetria and tremor), then therapists should seek alternative methods of evaluation of UE coordination. PMID- 8421721 TI - The effect of frequency of kinetic feedback on learning an isometric force production task in nondisabled subjects. AB - BACKGROUND AND PURPOSE: The purpose of this study was to investigate the effects of kinetic feedback frequency and concurrent kinetic feedback on the performance and learning of an isometric force production task in young, nondisabled adults. SUBJECTS: Twenty-four nondisabled, right-hand-dominant adults (18 male, 6 female), aged 19 to 33 years (mean = 22.5, SD = 4.1), participated in this study. METHODS: Eight subjects in each of three kinetic feedback groups performed an isometric elbow extension task in an attempt to minimize error between their effort and a force template over a 5-second period. Feedback was provided (1) concurrently with and after each attempt (concurrent feedback), (2) after each attempt (100% feedback), or (3) after every other attempt (50% feedback). Immediate and delayed (48-hour) retention tests were performed without feedback. Separate analyses of variance for repeated measures were used to compare task error among the three feedback groups for acquisition, immediate retention, and delayed retention trials. RESULTS: A significant interaction was found during the acquisition trial blocks, but at each trial block, subjects in the concurrent feedback group exhibited less error than did the subjects in either the 50% or 100% feedback group during the acquisition trials. For the immediate retention test, the 50% and 100% feedback groups exhibited 58% and 39% less error, respectively, than did the concurrent group. For the delayed retention test, the 50% and 100% feedback groups exhibited 52% and 26% less error, respectively, than did the concurrent group. In the immediate and delayed retention tests, subjects in the 50% feedback group displayed less error (31% and 36%, respectively) than did the 100% feedback group. CONCLUSION AND DISCUSSION: When permanent changes in the performance of a motor task are desired, concurrent feedback about task performance may be less desirable than feedback that is provided after the task has been completed. In addition, when feedback was used after the task had been completed, a lower frequency of feedback resulted in more permanent changes in the subjects' ability to complete the task. PMID- 8421722 TI - Alternative approaches to research in physical therapy: positivism and phenomenology. AB - This article presents philosophical approaches to research in physical therapy. A comparison is made to demonstrate how the research purpose, research design, research methods, and research data differ when one approaches research from the philosophical perspective of positivism (predominantly quantitative) as compared with the philosophical perspective of phenomenology (predominantly qualitative). Differences between the two approaches are highlighted by examples from research articles published in Physical Therapy. The authors urge physical therapy researchers to become familiar with the tenets, rigor, and knowledge gained from the use of both approaches in order to increase their options in conducting research relevant to the practice of physical therapy. PMID- 8421723 TI - Imaging of the pleura. PMID- 8421724 TI - A la recherche du temps perdu and the thymus (with apologies to Marcel Proust) PMID- 8421725 TI - Storage phosphor radiography of the chest. PMID- 8421726 TI - How to measure carotid stenosis. PMID- 8421727 TI - MR imaging is highly sensitive for acute subarachnoid hemorrhage ... not! PMID- 8421728 TI - Noninvasive carotid imaging: caveat emptor. PMID- 8421729 TI - Noninvasive carotid evaluation: carpe diem. PMID- 8421730 TI - Carotid endarterectomy: preoperative evaluation of candidates with combined Doppler sonography and MR angiography. Work in progress. AB - Doppler sonography and magnetic resonance (MR) angiography were prospectively used in combination as a substitute for conventional angiography in 24 consecutive patients likely to undergo carotid endarterectomy. Of 19 patients (20 lesions) who underwent carotid endarterectomy, 18 had not undergone preoperative angiography. High-grade lesions (> 75%-diameter stenosis) were surgically confirmed in 16; a 60%-70% stenosis, in one; and subtotal occlusion of the internal carotid artery, in another. Angiography was performed in a case of suspected internal carotid artery dissection. In the five nonsurgical cases, no significant stenosis was noted with both techniques in two instances, total occlusion was seen in two cases, and an asymptomatic stenosis was seen in one. Follow-up (average, 4.5 months) showed one case of symptomatic occlusion of the internal carotid artery after endarterectomy. This preliminary study suggests that the combined use of Doppler sonography and MR angiography can, in up to 79% (15 of 19) of cases, replace angiography for the preoperative evaluation of patients likely to need carotid endarterectomy. PMID- 8421731 TI - Carotid artery: prospective blinded comparison of two-dimensional time-of-flight MR angiography with conventional angiography and duplex US. AB - A prospective blinded comparison of two-dimensional (2D) time-of-flight (TF) magnetic resonance (MR) angiography and color duplex flow ultrasound (US) with conventional angiography as a standard of reference was performed in 50 patients with hemispheric ischemic symptoms. The guidelines of the North American Symptomatic Carotid Endarterectomy Trial for measuring stenosis of the internal carotid artery were utilized for maximum-intensity-projection (MIP) images and conventional angiograms. While the 2D TF MIP technique overrepresented the degree of stenosis in the internal carotid arteries, it matched the performance of color duplex flow US as measured by means of receiver operating characteristic curves. A signal void on the MIP images corresponded to a 70% or greater internal carotid artery stenosis in 17 of 20 arteries. At its current state of development, the accuracy of 2D TF MR angiography equals that of US in characterizing the degree of carotid stenosis but cannot be considered a replacement for conventional angiography. PMID- 8421732 TI - Subarachnoid hemorrhage: evaluation with MR imaging. AB - Thirty-seven magnetic resonance (MR) examinations were performed at 0.5 T in 33 patients with subarachnoid hemorrhage (SAH) caused by a ruptured aneurysm. Images were obtained 2 hours to 75 days after the ictus. Twenty-four proton-density weighted (long repetition time [TR], short echo time [TE]) images were obtained in the acute stage (< 72 hours after the ictus) of SAH; SAH was hyperintense to brain parenchyma and cerebrospinal fluid in all cases. The detectability of acute SAH on T1- (short TR, short TE) and T2- (long TR, long TE) weighted images was 36% and 50%, respectively. In the subacute and chronic stages (> 3 days after the ictus), the detectability of SAH on T1-, T2-, and proton-density-weighted images was 73%, 31%, and 83%, respectively. Although computed tomography is still the modality of choice for evaluating acute SAH, the authors emphasize that even acute SAH can be reliably demonstrated with MR imaging with the appropriate parameters. PMID- 8421733 TI - Measurement of brain activity with bolus administration of contrast agent and gradient-echo MR imaging. AB - This study was performed to measure changes in cerebral blood volume (CBV) associated with visual activation by use of bolus administration of contrast agent and conventional, clinically configured magnetic resonance (MR) hardware and software. Fast gradient-recalled acquisition in the steady state technique was used to study five healthy subjects during visual activation and a control dark state. MR images were obtained every 2.048 seconds for 2 minutes. A bolus of gadopentetate dimeglumine was injected during visual stimulation and darkness. Cine images produced from the series of rapid images clearly depicted arterial, capillary, and venous phases. Analysis of serial concentration maps derived from the rapid images revealed expected differences between the relative CBV of gray matter and that of white matter, as well as significantly increased relative CBV in calcarine cortex during visual activation versus the control state (mean increase, 15.24%; range, 6.41%-27.78%; P < .05). These results confirm those reported in echo-planar imaging studies and demonstrate that brain function can be assessed with the bolus method by means of MR imaging hardware and software with conventional clinical configurations. PMID- 8421734 TI - Posterior lobe of the pituitary in diabetes insipidus: dynamic MR imaging. AB - Magnetic resonance (MR) imaging was performed in 10 subjects: four with central idiopathic diabetes insipidus (DI), two with nephrogenic DI, and four with normal pituitary glands. Characteristics high intensity of the posterior pituitary lobe was seen in all four control subjects and one case of nephrogenic DI. However, it was undetectable in all four cases of central idiopathic DI and one case of nephrogenic DI. Peak contrast enhancement of the posterior lobe occurred within 30 seconds after injection of gadopentetate dimeglumine in control subjects and in both cases of nephrogenic DI and at 30-60 seconds in one case of mild partial idiopathic DI. In contrast, no or faint early enhancement of the posterior pituitary lobe was observed in two cases of complete and one case of partial central DI. In patients with central DI, MR imaging reveals delayed enhancement of the posterior pituitary lobe compared with the early enhancement seen in subjects without central DI. PMID- 8421735 TI - Brain blood flow in the dementias: SPECT with histopathologic correlation. AB - Single photon emission computed tomography (SPECT) of regional cerebral blood flow (RCBF) has been employed experimentally in the assessment of patients with dementia. The standard with which the SPECT diagnosis has been compared previously has been the initial clinical diagnosis. Recognizing that histopathologic diagnosis would be a more reliable standard, the authors compared SPECT diagnoses and clinical diagnoses with histopathologic diagnoses in a series of 18 patients who had been referred by the Alzheimer Disease Research Center. SPECT RCBF studies were carried out prospectively in 15 patients with an inhaled xenon-133 SPECT technique and in three patients with technetium-99m hexamethyl propylene-amine oxime and triple-camera-scanner SPECT. When compared with histopathologic diagnosis, clinical diagnosis was correct in 15 of 18 patients; visual scanning diagnosis, in 13 of 18; and Xe-133-SPECT diagnosis based on quantitative ratios in regions of interest, in 14 of 15 (13 of 13 with Alzheimer disease). PMID- 8421736 TI - Complete obstruction of the nasolacrimal system. Part I. Treatment with balloon dilation. AB - Fluoroscopically guided balloon dilation in treatment of complete obstruction of the nasolacrimal system was assessed in 39 eyes of 35 patients. All patients had severe epiphora. With nasal endoscopic guidance, a 0.018-inch guide wire was introduced through the superior punctum into the inferior meatus of the nasal cavity and was pulled out through the nasal aperture with a hemostat. A deflated angioplasty balloon catheter was passed retrograde over the guide wire through the inferior meatus until it straddled the stricture. Balloons were dilated with water-soluble contrast medium. No major complications occurred. At 7 days after balloon dilation, epiphora decreased in 22 of 39 eyes; 17 demonstrated complete resolution of severe epiphora, and five showed incomplete resolution. At 2 months, improvement had not been maintained in 10 of the 22 eyes. This technique for treatment of complete obstruction of the nasolacrimal system is simple and safe, but the high failure and recurrence rates in this study are not encouraging. PMID- 8421737 TI - Complete obstruction of the nasolacrimal system. Part II. Treatment with expandable metallic stents. AB - To evaluate the clinical usefulness of expandable metallic stents for maintenance of the dilated nasolacrimal sac and duct, double stents were implanted under fluoroscopic guidance in seven patients with severe epiphora after failed balloon dacryocystoplasty. Stents were constructed of 0.25-mm stainless steel wire in a cylindrical zigzag configuration of 10 bends. A single stent was 4 mm in diameter when fully expanded and 10 mm long. The stent was placed accurately in all but one patient, in whom a second stent was placed accurately after removal of the dislodged stent with a hemostat under endoscopic guidance. After stent placement, all patients demonstrated complete resolution of epiphora. During follow-up of 4 20 weeks, complete blockage of the stent occurred in one patient. This was treated with intrastent balloon dacryocystoplasty. No stents migrated. Although the long-term validity of this treatment has not been proved, expandable metallic stents seem to be of value in the treatment of complete obstruction of the lacrimal sac and nasolacrimal duct. PMID- 8421738 TI - Evaluation of the craniocervical junction in Down syndrome: correlation of measurements obtained with radiography and MR imaging. AB - Measurement of the atlas-dens interval is the radiographic standard for identification of patients with Down syndrome at high risk for neurologic injury from spinal cord compression. In 17 pediatric patients with Down syndrome, measurements of the atlas-dens interval, distance between the clivus and posterior odontoid process, and width of the neural canal obtained with plain radiographs were compared to predict the width of the subarachnoid space on magnetic resonance (MR) images. In all patients, cervical radiography and craniocervical MR imaging were prospectively performed during lateral flexion and extension. The width of the subarachnoid space was measured with MR imaging. When the 95% confidence interval for correlation coefficients of regression was used, subarachnoid space width on MR images correlated with neural canal width on radiographs better than with either atlas-dens interval or clivus-posterior odontoid process distance (P = .05). Measurement of neural canal width is a better predictor of potential spinal cord compression than the atlas-dens interval or clivus-posterior odontoid process distance and should be emphasized in screening examinations performed with plain radiography. PMID- 8421739 TI - Cervical trachea: dynamics in response to herniation of the normal thymus. AB - Nineteen infants aged 2 months to 2.5 years, first seen predominantly with stridor, were noted to have intermittent soft-tissue mass effect in the upper airway during routine evaluation with fluoroscopic or radiographic methods. The cervicothoracic trachea was always buckled posteriorly and, in almost all instances, to the right during forced exhalation (crying). Real-time ultrasound examination with the neck extended was used in these patients to define the cause of the tracheal dynamics. The intermittent cephalic movement of the homogeneous echotextured thymus from the anterior mediastinum into the neck was determined to be the probable cause of the mass effect in these infants. Magnetic resonance imaging in three infants confirmed this finding. The intermittent, physiologic suprasternal movement of the thymus in these infants did not by itself cause any luminal compromise of the trachea and did not result in any respiratory difficulty in these infants. PMID- 8421740 TI - Portable chest imaging: comparison of storage phosphor digital, asymmetric screen film, and conventional screen-film systems. AB - The quality of chest images obtained with portable radiography was evaluated for a conventional screen-film system, a new asymmetric screen-film chest system, and computed radiography (CR). Sixty chest images were obtained in 20 patients in an intensive care unit. The CR system was ranked by all three evaluating radiologists as substantially better in overall diagnostic quality, interpretability of the lungs, and musculoskeletal detail and by two of the three observers as better for the visibility of catheters and lines. In the upper abdomen and mediastinum, there was not a clear preference. Standard deviations of film density were +/- 0.12, +/- 0.41, and +/- 0.39 for the CR, conventional, and asymmetric systems, respectively. For the same systems, phantom results indicated the relative lung contrast values were 1.2, 1.0, and 0.89, respectively. Similarly, the limiting resolution values in the lung were 2.0, 4.2, and 6.3 line pairs per millimeter. The CR system had twice the root-mean-square noise of the screen-film systems. Overall, the preferred system for portable chest imaging was the CR system. PMID- 8421741 TI - Digital chest radiography with photostimulable storage phosphors: signal-to-noise ratio as a function of kilovoltage with matched exposure risk. AB - A photostimulable storage phosphor (PSP) digital radiography system was evaluated regarding the signal-to-noise ratio (S/N) on chest images acquired at differing peak kilovoltage settings but with matched risk from radiation exposure. Images of two chest phantoms were acquired by using bedside (portable) imaging equipment at tube voltages ranging from 60 to 120 kV. Exposure factors were set so that the effective dose equivalent, a risk estimator weighted for various organs, was approximately equal in all exposures. The S/N in the lung-equivalent regions was found to be slightly higher (maximum, 15%) in the low-energy exposures, while the S/N values in the mediastinum- and subdiaphragm-equivalent regions were approximately equal at all kilovoltage settings. The absence of a high sensitivity of S/N to kilovoltage in risk-matched PSP images should enable institutions to select x-ray beam quality on the basis of other imaging criteria. PMID- 8421742 TI - Computed equalization radiography: preliminary clinical evaluation. AB - Computed Equalization Radiography (CER) was designed to increase visualization of mediastinal structures without affecting lung contrast. CER optimizes the image by obtaining two 1-second fan beam scans. The initial low-dose prescan determines the location of the lungs by means of a stationary krypton gas detector. During the equalization scan, this information is used to position 35 beam attenuators in real time so as to increase exposure of the mediastinum while maintaining ideal optical density over the lungs. Eight observers analyzed 20 pairs of posteroanterior (PA) and lateral radiographs obtained on conventional and CER units. On the PA images, three of four mediastinal areas analyzed were visualized better in CER images, while there was no difference in lung detail. There were no appreciable differences in the lateral projection between the CER and conventional images. Contrast on the PA radiograph was preferred on the CER image. CER provided modest improvement in visualization of mediastinal structures and maintained lung contrast without degrading lung detail on the PA image. CER had no major effect on visualization in the lateral images. PMID- 8421743 TI - Determining the likelihood of malignancy in solitary pulmonary nodules with Bayesian analysis. Part I. Theory. AB - Only two radiographic findings allow reliable distinction of benign from malignant solitary pulmonary nodules. Intuitively, it is clear that other radiographic and clinical findings should also be important in making this distinction. Subjectively incorporating these other findings into the decision of whether a nodule is benign or malignant is difficult. Likelihood ratios, which indicate the degree of malignancy or benignity represented by a test result or clinical finding, can be combined by means of the Bayes theorem to quantitate the probability of malignancy of a given nodule. From a literature survey, likelihood ratios were derived for six radiographic and four clinical characteristics associated with solitary pulmonary nodules. There were a total of 15 malignant and 19 benign findings, the most important of which were radiographic characteristics. For malignant nodules, the most important radiographic characteristics were thickness of the cavity wall spicular edge, and diameter of over 3 cm. For benign nodules, the most important radiographic characteristics were benign growth rate and a benign pattern of calcification. PMID- 8421744 TI - Determining the likelihood of malignancy in solitary pulmonary nodules with Bayesian analysis. Part II. Application. AB - Four board-certified radiologists estimated the probability of malignancy in 66 cases of solitary pulmonary nodules. Two other radiologists evaluated the same nodules according to various radiographic and clinical findings. These findings were then used to estimate the probability of malignancy by using previously derived likelihood ratios and the Bayes theorem. The readers using Bayesian analysis performed significantly better than the expert readers (P < .05) when individual radiographs were considered and when all radiologic studies were combined. In addition, the readers using Bayesian analysis misclassified fewer malignant nodules as benign (mean, 6.5) than did the expert readers (mean, 6.5) than did the expert readers (mean, 16.5). The authors conclude that Bayesian analysis may be a useful aid in the evaluation of solitary pulmonary nodules. PMID- 8421745 TI - Mediastinal bronchogenic cyst: demonstration of a fluid-fluid level at MR imaging. AB - The appearance of mediastinal bronchogenic cysts at cross-sectional imaging is variable. Intracystic hemorrhage, proteinaceous fluid, or suspended calcium may belie the true cystic nature of the lesion at computed tomography or magnetic resonance (MR) imaging. The authors report a case of a fluid-fluid level seen on MR images of a bronchogenic cyst. This appearance, likely due to dependent layering of proteinaceous material, helped preoperatively confirm the true cystic nature of the lesion. PMID- 8421746 TI - New high-resolution teleradiology system: prospective study of diagnostic accuracy in 685 transmitted clinical cases. AB - To compare the accuracy of interpretation of digitized radiographs with that of plain films, the authors prospectively evaluated the first 685 plain film cases (530 adult and 155 pediatric cases, each of which consisted of one or more images) transmitted from an outpatient center 18 miles (approximately 29 km) to a hospital radiology department by means of a high-speed teleradiology system. Plain films were digitized and transmitted via a T-1 (1.544 Mbit/sec) data link for display on high-resolution (2,560 x 2,048-pixel) workstations. Radiologists at the hospital used a copy of the radiology requisition that had been faxed from the remote center. Interpretation of the digital images was followed by review of the original plain radiographs within 1 working day. Discrepant interpretations occurred in 18 cases (2.6%) (sensitivity, 96%; specificity, 99%); an arbitration panel decided that they were associated with observer performance more than with the fidelity of the digital display. It is concluded that primary diagnosis without review of the original plain radiographs is feasible with state-of-the art teleradiology systems. PMID- 8421747 TI - The normal shoulder: common variations that simulate pathologic conditions at MR imaging. AB - The appearance of the supraspinatus tendon and anterior capsular mechanism was analyzed in 60 asymptomatic shoulders with magnetic resonance (MR) imaging. The images were reviewed with special attention to findings that simulate pathologic conditions, as defined by means of currently accepted criteria. On T1-weighted and proton-density (PD) spin-echo (SE) images, intermediate signal intensity was present within the supraspinatus tendon in most shoulders. Focal signal intensity within the distal tendon was particularly common finding, being present in 95% (57 of 60) of shoulders on PD images. Focal obliteration of the subacromial subdeltoid fat stripe and acromioclavicular joint arthrosis were seen in 95% (57 of 60) and 48% (29 of 60) of the subjects, respectively. There was considerable variation in the shape of the anterior glenoid labrum-glenohumeral ligament (GHL) complex. The labrum may appear triangular, round, crescentic, or absent. The middle and inferior GHLs lie in proximity to the upper half of the anterior labrum; the cleavage plane between the ligaments and the labrum can mimic a tear at MR imaging. PMID- 8421748 TI - Shoulder after surgery: MR imaging with surgical validation. AB - To assess the accuracy of magnetic resonance (MR) imaging in the evaluation of the shoulder after surgery, MR examinations were performed in 31 patients before repeated surgery, and MR findings were correlated with the subsequent operative findings. In addition, the MR findings associated with prior surgery were reviewed, including altered structure of the acromion, soft-tissue metal artifacts, a surgical trough in the humeral head, nonvisualization of the subacromial-subdeltoid fat stripe, and intermediate signal intensity within the substance of the rotator cuff on images obtained with all pulse sequences. The MR criteria for full-thickness tears in the shoulder after surgery are the presence of fluidlike signal intensity on T2-weighted images that extends through an area of the rotator cuff or the nonvisualization of a portion of the rotator cuff. With use of these criteria, six of seven full-thickness rotator cuff tears were identified at MR imaging, with an accuracy of 90%. Partial cuff tears were indistinguishable from repaired tendons. Findings at MR imaging were 74% accurate concerning the presence of impingement. PMID- 8421749 TI - Two segments of the supraspinous muscle: cause of high signal intensity at MR imaging? AB - A zone of increased signal intensity seen in asymptomatic subjects near the insertion of the supraspinous tendon onto the greater tuberosity ("pseudogap") leads to difficulty in the interpretation of magnetic resonance (MR) images of the shoulder. To better understand this pseudogap, the authors dissected and sectioned the shoulders of four cadavers and analyzed 20 MR images of patients' shoulders. They found that the supraspinous muscle consists of two distinct portions: an anterior fusiform portion that contains the dominant tendon and a straplike posterior portion. The orientation of the dominant tendon differs from that of the main muscle by approximately 10 degrees. In addition, oblique-coronal MR images were obtained parallel to the supraspinous main muscle and the central tendon in six volunteers; the appearance of the pseudogap was similar with both projections. Fat-saturated and gradient-echo images of the shoulder were obtained with increasing echo times in four additional volunteers. There was no evidence of fat within the pseudogap. Because the anatomic composition of the pseudogap could not be explained with partial volume averaging with adjacent muscle or fat, the authors conclude that it may represent a manifestation of unique focal tissue relaxation parameters or depend on the tendon orientation in the main magnetic field. PMID- 8421750 TI - Liposarcoma of the extremities: MR and CT findings in the histologic subtypes. AB - The findings on images of liposarcomas of the extremities in 48 patients (26 men and 22 women aged 20-85 years) were reviewed to correlate the histologic subtypes with radiologic appearance. Computed tomographic scans were obtained in 36 patients; magnetic resonance (MR) images, in 27 patients. The study group had 19 myxoid, 12 well-differentiated, nine round cell, and eight pleomorphic liposarcomas. Well-differentiated liposarcomas were predominantly composed of fat, typically with thick septa, which were hyperintense on T2-weighted spin-echo MR images. The heterogeneity of tumor helped differentiate tumor subtypes. Eleven of 19 myxoid tumors were mildly heterogeneous. Round cell and pleomorphic subtypes, which tend to be aggressive, were generally heterogeneous. Well differentiated liposarcomas may be confidently differentiated from other subtypes of liposarcoma. Except for the well-differentiated subtypes, liposarcomas often contain no fat. Moderate to marked heterogeneity is common in high-grade liposarcomas; myxoid liposarcomas tend to be homogeneous and may mimic cysts. PMID- 8421751 TI - Rapidly destructive hip disease: clinical and imaging abnormalities. AB - The clinical and radiographic records of 23 patients (15 women, eight men) with rapidly destructive hip disease (RDHD) were retrospectively reviewed. Criteria for RDHD included a history of hip pain of 1-6 months duration and the radiographic appearance of a rapidly progressive atrophic form of bone destruction involving both the femoral head and the acetabulum. Radiographs of the remainder of the appendicular skeleton were assessed in 14 patients. The mean patient age was 72 years. The average time from clinical presentation to the appearance of severe hip destruction was 14 months. Five patients demonstrated similar atrophic bone destruction around other articulations. No patients had clinical or laboratory evidence of sepsis or neurologic disease. Although previous reports have suggested that RDHD is degenerative in nature, similar involvement of other articulations suggests that it may represent a focal finding of a more generalized process. PMID- 8421752 TI - Regional variations in bone mineral density as assessed with dual-energy photon absorptiometry and dual x-ray absorptiometry. AB - Total-body, lumbar spine, and hip bone mineral density (BMD) scans were obtained in postmenopausal women to examine regional variations in mineralization of trabecular bone. One hundred ninety-nine patients were studied with dual-energy photon absorptiometry (DPA) and/or dual x-ray absorptiometry (DXA). Comparison of BMD at the different sites showed statistically significant correlations (P < .001) among all the sites; however, variations between sites were observed in many patients. Total-body measurements were within normal limits as defined by the system software in 90% of patients with substantial bone mineral loss in the spine or hip. Lumbar spine measurements calculated from the total-body scan differed from the direct measurements by an average of 10% (DPA) and 12% (DXA). These results suggest that site-specific measurements are required to assess regional osteopenia. While total-body scans may be precise and offer the advantage of total-body composition determination, BMD values derived from total body scans cannot currently replace direct measurements. PMID- 8421753 TI - Computerized radiographic analysis of osteoporosis: preliminary evaluation. AB - Measurement of bone mass is important in determining the risk for fracture and in following the course of patients undergoing therapy for osteoporosis. Bone mineral densitometry (BMD) is a good predictor of fracture risk, but there is considerable overlap in BMD measurements between individuals with fracture risk and those without. In this study, computerized texture analysis of the trabecular pattern on conventional spine radiographs was used to evaluate bone structure as a determinant of fracture risk. Standard lumbar spine radiographs of 43 individuals were analyzed and compared with BMD measurements obtained with dual photon absorptiometry. This method was more effective than BMD in differentiation of patients with fractures elsewhere in the spine from those with no fracture. These preliminary results suggest that this method of bone structure analysis, combined with BMD, may lead to a more sensitive and specific predictor of osteoporosis and risk of fracture. PMID- 8421754 TI - Duchenne muscular dystrophy: MR grading system with functional correlation. AB - The pelvic girdle, thigh, and calf muscles of 29 patients with Duchenne muscular dystrophy (DMD) were evaluated with magnetic resonance (MR) imaging. The muscles that were most resistant to disease were the gracilis, followed by the sartorius, semitendinous, and semimembranous. Sixteen patients had asymmetric involvement in part of the muscle bundles of thigh. An MR grading system was developed to assess the number of preserved pelvic and thigh muscles, the severity of fatty infiltration of the calf, and the increase of subcutaneous fat. Statistically significant correlations were obtained between MR grade and clinical functional grade (P < .01), MR grade and disease duration (P < .01), and MR grade and patient age (P < .01). An inverse correlation was observed between the creatine kinase values and the MR grade (P < .05). The MR grading system helped identify disease severity in patients with low clinical functional grades. Twenty-four follow-up studies were performed in 22 patients. In 13 of the 24 cases, the MR grade progressed while the clinical functional grade remained unchanged. Both the MR and clinical functional grades progressed in six cases. The results suggest that MR imaging may be useful in prebiopsy mapping and may help accurately monitor the progression of DMD. PMID- 8421755 TI - Teratomas versus cystic hemorrhagic adnexal lesions: differentiation with proton selective fat-saturation MR imaging. AB - To assess the value of proton-selective fat-saturation magnetic resonance (MR) imaging in differentiating teratomas from cystic hemorrhagic masses, 38 patients with 48 lesions (21 teratomas, 27 cystic hemorrhagic masses) detected at prior ultrasound or computed tomography were imaged with standard T1- and T2-weighted spin-echo (SE) sequences. Twenty-one patients with 28 lesions (13 teratomas, 15 cystic hemorrhagic masses) were imaged with an additional T1-weighted fat saturation sequence. On standard T1- and T2-weighted SE images, 43% of the teratomas and 52% of the cystic hemorrhagic masses were characterized correctly with signal intensity criteria. Sixty-two percent of the teratomas and 100% of the cystic hemorrhagic masses were characterized correctly with chemical shift artifact criteria. With fat-saturation images alone, the characterization sensitivity for teratomas and cystic hemorrhagic masses increased to 92% and 100%, respectively. Fat-saturation MR imaging was statistically superior to standard T1- and T2-weighted imaging in characterizing teratomas. PMID- 8421756 TI - Characterization of hemorrhagic adnexal lesions with MR imaging: blinded reader study. AB - To assess the performance of published criteria for differentiation of endometriomas from other hemorrhagic adnexal lesions with magnetic resonance (MR) imaging, 74 lesions with pathologic proof in 46 patients were evaluated by means of a 1.5-T system with use of conventional T1-weighted and either T2-weighted spin-echo (SE) or fast SE sequences. The MR images were independently presented to each of three blinded readers, and receiver operating characteristic (ROC) curve analysis was performed. The criterion of a cyst with high signal intensity on short repetition time (TR)/echo time (TE) images and shading on long TR/TE images had a mean sensitivity of 68%, mean specificity of 83%, and mean accuracy of 76% for diagnosis of endometrioma. Ancillary findings previously reported to be suggestive of endometrioma (eg, low-signal-intensity rims, adhesions, bowel tethering, and implants) were proved inaccurate and showed low interobserver agreement. ROC analysis showed similar area measurements among the three radiologists for diagnosis of endometrioma. MR imaging had only moderate accuracy in distinction of endometriomas from other hemorrhagic adnexal lesions. PMID- 8421757 TI - Normal uterus and FIGO stage I endometrial carcinoma: dynamic gadolinium-enhanced MR imaging. AB - Dynamic magnetic resonance (MR) imaging was performed on a 1.5-T superconducting unit for evaluation of 26 stage I endometrial carcinomas. To establish the appearance of the normal uterus, 27 normal uteri were also evaluated. After rapid injection of gadopentetate dimeglumine, dynamic images were obtained every 30 seconds with the spin-echo technique in the sagittal plane. On dynamic studies of endometrial carcinoma, the tumor-myometrial contrast was marked at 120 seconds after administration of gadopentetate dimeglumine (contrast-to-noise ratio [C/N], 26.0). The tumor-myometrial contrast on the dynamic study was more marked than that on postcontrast T1-weighted images (C/N, 10.0) and on T2-weighted images (C/N, 2.14). Dynamic and postcontrast MR images were superior in enabling differentiation of viable tumors from necrosis or residual secretion in the endometrial cavity. In the evaluation of presence of tumor and myometrial invasion, the accuracy of T2-weighted imaging and dynamic imaging was 67.9% and 84.9%, respectively. PMID- 8421758 TI - Coaxial percutaneous biopsy technique with automated biopsy devices: value in improving accuracy and negative predictive value. AB - Three hundred percutaneous biopsies were performed in 267 consecutive patients by means of a coaxial technique with use of 18- and 20-gauge automated cutting biopsy devices and 22-gauge aspiration needles. Thoracic, hepatic, renal, pancreatic, adrenal, splenic, retroperitoneal, and musculoskeletal soft-tissue masses were sampled. For malignant masses (229 cases), the sensitivity was 79% for cytologic analysis, 88% for histologic analysis, and 92% for both combined. In benign disease (71 cases), a correct specific diagnosis was made with cytologic analysis in 38%, with histologic analysis in 97%, and with both combined in 97%. The negative predictive value was 60% for cytologic analysis, 72% for histologic analysis, and 80% for both combined. When only cancer-negative results in which a specific benign diagnosis was made were considered, the negative predictive value was 100% for cytologic analysis, 97% for histologic analysis, and 97% for both combined. The positive predictive value was 100% for both cytologic and histologic analysis. Bleeding complications occurred in 3% of biopsies, including in one patient who died. PMID- 8421759 TI - Transjugular intrahepatic portosystemic shunts in patients with portal vein occlusion. AB - The feasibility and efficacy of transjugular intrahepatic portosystemic shunts (TIPS) were evaluated in 10 patients with preexisting portal vein occlusions. A standard transjugular approach was used in six of the 10 patients for both portal vein recanalization and TIPS placement. The protal veins were successfully recanalized and TIPS were established in three of the six patients. TIPS placement was unsuccessful in the other three patients because the catheters could not be advanced through the occluded segments. A transhepatic approach was used in four of the 10 patients for portal vein recanalization before transjugular catheterization and TIPS placement were attempted. Both portal vein recanalization and TIPS placement were technically successful in all four patients. Bleeding stopped in all patients after successful shunt placement. TIPS can be used to control variceal bleeding in some patients, despite preexisting portal vein occlusion. Preliminary recanalization of the occluded portal segment by means of the transhepatic approach may facilitate TIPS placement. PMID- 8421760 TI - Transjugular intrahepatic portosystemic shunt: evaluation with Doppler sonography. AB - To assess the potential role of ultrasound (US) in evaluating transjugularly inserted intrahepatic portosystemic shunts, the authors performed sonographic, including duplex and color Doppler, studies in 23 patients. Imaging was performed before, during the first week after, and 2-3 months after shunt placement. The prostheses were readily visualized and shunt patency was easily determined with Doppler US in all patients. The mean of the maximum blood flow velocity in the main portal vein increased from 7 (range, 3-16) cm/sec before shunting to 24 (range, 18-47) cm/sec 2-3 months after shunting. Flow reversal was detected in the intrahepatic portal branches in 16 (69%) of the 23 patients. US allowed prompt detection and management of two cases of incomplete stent expansion and one case of shunt stenosis due to pseudo-intimal hyperplasia. Sonography is a valuable tool for evaluation of changes in hepatic hemodynamics after transjugular intrahepatic shunt placement. PMID- 8421761 TI - Echo-planar MR imaging of normal and ischemic myocardium with gadodiamide injection. AB - Rapid echo-planar (EP) magnetic resonance (MR) imaging was used to monitor the first pass of a bolus of gadodiamide injection in the hearts of normal rats and rats subjected to left coronary artery occlusion. Inversion-recovery EP imaging combined with a low dose (0.05 mmol/kg) of the contrast agent caused signal enhancement of normal myocardium from 19% +/- 4 to 63% +/- 5 (mean +/- 1 standard error of the mean) of fully relaxed intensity at the peak of the bolus but only slight increase in signal intensity of the ischemic zone. Thus, ischemic myocardium was demarcated as a hypointense zone (cold spot) during passage of the bolus. A higher dose (0.20 mmol/kg) of the same agent caused signal loss of normal myocardium from 100% to 39% +/- 7 of control at the peak of the bolus on gradient-recalled echo EP images, and ischemic myocardium was visualized as a hyperintense zone (hot spot). With either method of monitoring bolus transit, myocardial signal intensity recovered slowly following the peak bolus effect, consistent with substantial extraction of the agent during the first pass through the heart. Use of gadodiamide injection can allow discrimination between ischemic and nonischemic myocardium on both T1- and susceptibility-weighted EP images during bolus transit. PMID- 8421762 TI - Orally administered manganese chloride: enhanced detection of hepatic tumors in rats. AB - To evaluate its potential as a tissue-specific hepatobiliary magnetic resonance (MR) contrast agent, manganese chloride was orally administered to rats in increasing doses of 100-1,500 mumol/kg MnCl2, and the relaxation times of liver, pancreas, kidney, and heart were measured with ex vivo relaxometry and in vivo MR imaging. Two hours after ingestion of 200 mumol/kg MnCl2, liver T1 was decreased by 48%, whereas tumor T1 decreased by only 9%. On spin-echo MR images, the signal to-noise ratio in liver increased by 54%; the contrast-to-noise ratio in tumor, by 375%. The T1 of pancreas, kidney, and heart decreased by 8%, 23%, and 13%, respectively. At subjective assessment, the signal intensity of the upper gastrointestinal tract was reduced, likely because of the high concentration of manganese in the lumen, and delineation of the intestine from other abdominal structures was improved. These results indicate that orally administered MnCl2 causes substantial, reproducible, and tissue-specific enhancement of liver. Because enhancement of tumor was minimal, orally administered MnCl2 may potentially be used to improve detection of hepatic tumors. PMID- 8421763 TI - Fine-needle biopsy: prospective comparison of aspiration versus nonaspiration techniques in the abdomen. AB - A prospective study was designed to compare the aspiration (suction method) and nonaspiration (nonsuction method) techniques of fine-needle biopsy (FNB) in 50 consecutive patients with abdominal pathologic conditions. Sites of biopsy included liver (n = 24), retroperitoneum (n = 9), adrenal gland (n = 5), pancreas (n = 4), omentum (n = 4), and miscellaneous sites (n = 4). Aspiration and nonaspiration FNBs were performed in each lesion with 22-gauge needles, and results were interpreted by a single cytopathologist. Cytologic specimens obtained with each technique were analyzed for diagnostic accuracy, total number of cell clusters per biopsy (graded 0-10, 10-20, 20-30, and > 30), presence of crush artifact, and amount of blood present (graded from 0 to +3). No significant differences were seen between the aspiration and nonaspiration techniques with regard to number of cell clusters per biopsy (44 of 50 specimens vs 42 of 50) (P < .0003), amount of blood present (grade 2.3 vs 2.2) (P < .0003), and amount of crush artifact. The positive predictive value for the aspiration technique was 91.5% versus 74% for the nonaspiration technique. The aspiration technique of FNB appears superior to the nonaspiration technique in the abdomen. PMID- 8421764 TI - Retrogastric colon: CT demonstration of anatomic variations. AB - In a series of 9,400 body computed tomographic (CT) scans of adult patients, a retrogastric course of the transverse colon with interposition of colon between the pancreatic body-tail and stomach was identified in 18 patients (0.2%). Three other patients (0.03%) had retrosplenic, retrogastric colon with transverse colon interposed between the spleen and posterior diaphragm. Three of the 18 patients with pancreaticogastric interposition also had small bowel malrotation. Clinically, no symptoms attributable to retrogastric colon were found. Retrogastric colons probably result from mild embryologic abnormalities of bowel rotation and fixation. Radiologically, colon in the area of the lesser sac must be differentiated from the normal duodenojejunal flexure, a perforated viscus, and abscess. PMID- 8421765 TI - Malignant stricture with colovesical fistula: stent insertion in the colon. AB - The authors describe a 91-year-old man with a malignant stricture of the sigmoid colon complicated with a colovesical fistula, which was successfully managed with insertion of a metallic esophageal stent. The fistula closed, and the stent function was good during the observation time of 1 month. Temporary stent placement for treatment of malignant strictures in the large bowel is proposed. PMID- 8421766 TI - Long-term results of curative irradiation in pathologically staged IA and IIA Hodgkin disease. AB - One hundred seventy-six patients with pathologically staged IA and IIA Hodgkin disease (HD) treated with irradiation alone were evaluated for long-term survival and freedom from relapse (FFR). Most of the patients received treatment to mantle and paraaortic fields; chemotherapy was not given except as salvage therapy. For pathologically staged IA disease, the 5-, 10-, and 15-year survival rates were 94%; the corresponding FFR rates were 96%, 93%, and 93%. For pathologically staged IIA disease, respective survival rates were 93%, 89%, and 80%, with FFR rates of 86%, 84%, and 84%. Twenty-one patients (12%) had relapse of HD; salvage therapy was successful in 11 of these patients. Pelvic recurrence was uncommon (three of 176 cases [2%]). No patient developed leukemia, and only two patients developed second malignancies (lung cancer in both cases). The authors conclude that radiation therapy is effective in treatment of early-stage HD. PMID- 8421767 TI - Management of oligodendrogliomas. AB - To define optimum treatment and delineate recurrence patterns and prognostic factors in oligodendrogliomas, the authors reviewed the records of 49 patients who received treatment for these rare tumors between 1957 and 1990; 41 patients had at least 5 years follow-up. Age, race, gender, performance of computed tomography (CT), tumor location and grade, histologic findings, calcifications, surgery with or without postoperative radiation therapy, and the doses and fields used in radiation therapy were evaluate for prognostic relevance by means of chi 2 and Wilcoxon log-rank tests. Metaanalysis of relevant literature was also performed. Actuarial survival at 5, 10, and 15 years was 61%, 41%, and 24%, respectively. The pattern of recurrence was predominantly local alone (30 of 31 patients [97%] with recurrence). Age younger than 40 years, low-grade tumor, calcifications, and performance of CT were statistically significant prognostic factors. A trend existed toward improved 5-year survival in patients who underwent postoperative radiation therapy (P = .067); at stratification for subtotal resection, this improvement became significant (74% vs 25%, P = .019). Metaanalysis demonstrated a survival advantage for surgery with radiation therapy versus surgery only (56% vs 42%, P < .01). PMID- 8421768 TI - Renal biopsy: in vitro and in vivo comparison of a new automatic biopsy device and conventional biopsy systems. Work in progress. AB - A nondisposable full-cut biopsy gun with a disposable needle (system 1, prototype of system 2) and a disposable gun-needle combination (system 2) were compared with conventional needles and Tru-Cut-type biopsy systems in renal parenchymal biopsy procedures. In cadaveric kidneys, more glomeruli were harvested with system 1 than with four other biopsy systems. In native kidneys, more glomeruli were harvested and core quality was better with system 2 than with a Tru-Cut-type biopsy gun. PMID- 8421769 TI - Spiral CT arterial portography of the liver. AB - The enhancement characteristics of the liver, aorta, and hepatic vein were evaluated during computed tomography with arterial portography (CTAP) performed with spiral technique in 10 patients. The attenuation of the liver was nearly constant over the scanning interval (average attenuation, 168 HU +/- 16). The attenuation of the aorta increased by an average of 54 HU +/- 31, and the average attenuation of the hepatic vein was 250 HU +/- 34. These data indicate that high levels of liver parenchymal and vascular enhancement can be reliably obtained with CTAP performed with spiral technique. PMID- 8421770 TI - Acronymophobia. PMID- 8421771 TI - Propofol for sedation of pediatric patients. PMID- 8421772 TI - Proposal for a shared resource in clinical diagnostic radiology research. PMID- 8421773 TI - Quality assurance in mammography: the Michigan breast phantom test experience. PMID- 8421774 TI - [Isolation of Borrelia burgdorferi in the cerebrospinal fluid of 3 children with neurological involvement]. AB - Isolation of Borrelia burgdorferi from the CSF is relatively rare. The present report describes the first three isolations in Switzerland. Clinically, our first observation confirmed the frequent association of B. burgdorferi with peripheral facial paresis in children. The other two cases illustrate the variety of symptoms in neuro-borreliosis. In the first case the culture was positive after 6 weeks. The results of serologic tests (indirect immunofluorescence and ELISA) for detection of antibodies against B. burgdorferi were negative or non-significant in this child's serum. On the other hand, specific antibodies (IgG) were detected in the serum by western blot. Culture of the second CSF already showed Borrelia growth after 10 days. Immunofluorescence revealed high antibody titers (1/256) against B. burgdorferi in this patient's serum. IgG showed a weakly positive reaction in western blot. The reliability of this result was confirmed by isolation of Borrelia. In neither of the two CSF could intrathecal synthesis of specific antibodies be demonstrated. In the third case, however, immunofluorescence showed IgG antibody titers of 1/128 in the CSF and 1/512 in serum. Intrathecal synthesis of specific antibodies was demonstrated with an index of 13.4 (norm < 2). Western blot confirmed the specificity of the reactions observed with the serum and CSF IgG. Culture of CSF produced significant growth of Borrelia within 7 days. Protein profile and reactions with poly- and monoclonal antibodies confirmed that the three strains belonged to B. burgdorferi.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421775 TI - ['Cardiac ballet' with and without amiodarone]. AB - Two well documented cases of torsades de pointes under amiodarone therapy are discussed. This special form of ventricular tachycardia is also known as "twisting around the points" or "cardiac ballet". Our first case involves recurrent torsades de pointes manifesting almost 2 weeks following onset of amiodarone therapy. They were successfully treated by overpacing. In the second case the isolated torsade de pointes was probably not caused by the longlasting and successful use of amiodarone but reflected severe ischemic heart disease and slight hypokalemia. This was confirmed in the one year follow-up, as torsade de pointes was no longer present despite increased dosage of amiodarone. Amiodarone induced torsades de pointes is very rare. Most cases reported in the literature are poorly documented or were caused by other factors (electrolyte disturbances, other antiarrhythmic medication). If amiodarone has been administered as a matter of utmost necessity, other etiologies must be excluded or eliminated before amiodarone therapy is stopped when torsades de pointes occurs. Torsade de pointes due to amiodarone arises independently of the duration of therapy, the administered dosage and the extent of QT-prolongation. Holter monitoring does not allow a prediction and the benefit of electrophysiological studies is still controversial. The management of patients with amiodarone-induced torsades de pointes includes administration of magnesium and, frequently, overpacing. Due to the long half-life of amiodarone, prolonged temporary pacing may be required. General experience suggested that amiodarone can be used safely in patients who have developed torsades de pointes with other agents. PMID- 8421776 TI - [Retroperitoneal fibrosis and portal hypertension]. AB - A patient is described in whom retroperitoneal fibrosis manifested itself by bleeding from ruptured varices and pain in the left flank and lumbar region. The clinical diagnosis was confirmed by axial computerized tomography and MRI, while the histology from a mesenteric biopsy taken during splenectomy was considered insufficiently consistent with the diagnosis of retroperitoneal fibrosis. Treatment with prednisone resulted in relief of lumbar and flank pain a well as in prevention of further digestive tract hemorrhage. A review of the literature since 1966 shows 16 articles describing the association of retroperitoneal fibrosis and portal hypertension manifesting itself by hemorrhage from bleeding varices. PMID- 8421777 TI - [Proliferative myositis--a little-known pseudomalignant lesion]. AB - The leading symptom of proliferative myositis is a rapidly growing hard tumor in the muscle. The disease is rare and therefore widely unknown. We observed a case of proliferative myositis in a 54-year-old female. We initially interpreted the hard tumor in the left musculus sternocleidomastoideus and its rapid growth as a malignant disease. The histological findings finally led to the correct diagnosis. In the light of the literature of 78 reported cases, a presumptive diagnosis can already be suspected from the clinical examination. The addition of CT scan and fine needle aspiration is highly diagnostic. PMID- 8421778 TI - Gun ownership and risk. PMID- 8421779 TI - Antibiotic resistance. PMID- 8421780 TI - Gene therapy. Healy approves an unproven treatment. PMID- 8421781 TI - An inhibitor of p34CDC28 protein kinase activity from Saccharomyces cerevisiae. AB - The p34CDC28 protein from Saccharomyces cerevisiae is a homolog of the p34cdc2 protein kinase, a fundamental regulator of cell division in all eukaryotic cells. Once activated it initiates the visible events of mitosis (chromosome condensation, nuclear envelope breakdown, and spindle formation). The p34CDC28 protein also has a critical role in the initiation of DNA synthesis. The protein kinase activity is regulated by cycles of phosphorylation and dephosphorylation and by periodic association with cyclins. An endogenous 40-kilodalton protein (p40) originally identified as a substrate of the p34CDC28 protein kinase was purified. The p40 protein bound tightly to p34CDC28 and inhibited the activity of the kinase. The p40 protein may provide another mechanism to regulate p34CDC28 protein kinase activity. PMID- 8421782 TI - The prevention of thymic lymphomas in transgenic mice by human O6-alkylguanine DNA alkyltransferase. AB - Nitrosoureas form O6-alkylguanine-DNA adducts that are converted to G to A transitions, the mutation found in the activated ras oncogenes of nitrosourea induced mouse lymphomas and rat mammary tumors. These adducts are removed by the DNA repair protein O6-alkylguanine-DNA alkyltransferase. Transgenic mice that express the human homolog of this protein in the thymus were found to be protected from developing thymic lymphomas after exposure to N-methyl-N nitrosourea. Thus, transgenic expression of a single human DNA repair gene is sufficient to block chemical carcinogenesis. The transduction of DNA repair genes in vivo may unravel mechanisms of carcinogenesis and provide therapeutic protection from known carcinogens. PMID- 8421783 TI - Regulation of heat shock factor trimer formation: role of a conserved leucine zipper. AB - The human and Drosophila heat shock transcription factors (HSFs) are multi-zipper proteins with high-affinity binding to DNA that is regulated by heat shock induced trimerization. Formation of HSF trimers is dependent on hydrophobic heptad repeats located in the amino-terminal region of the protein. Two subregions at the carboxyl-terminal end of human HSF1 were identified that maintain the monomeric form of the protein under normal conditions. One of these contains a leucine zipper motif that is conserved between vertebrate and insect HSFs. These results suggest that the carboxyl-terminal zipper may suppress formation of trimers by the amino-terminal HSF zipper elements by means of intramolecular coiled-coil interactions that are sensitive to heat shock. PMID- 8421784 TI - Rate and mechanism of nonhomologous recombination during a single cycle of retroviral replication. AB - Oncogenes discovered in retroviruses such as Rous sarcoma virus were generated by transduction of cellular proto-oncogenes into the viral genome. Several different kinds of junctions between the viral and proto-oncogene sequences have been found in different viruses. A system of retrovirus vectors and a protocol that mimicked this transduction during a single cycle of retrovirus replication was developed. The transduction involved the formation of a chimeric viral-cellular RNA, strand switching of the reverse transcription growing point from an infectious retrovirus to the chimeric RNA, and often a subsequent deletion during the rest of viral DNA synthesis. A short region of sequence identity was frequently used for the strand switching. The rate of this process was about 0.1 to 1 percent of the rate of homologous retroviral recombination. PMID- 8421785 TI - Circadian rhythm in membrane conductance expressed in isolated neurons. AB - Although isolated neurons can generate rhythmic activity, they have not yet been shown to generate rhythms with a period in the circadian range (near 24 hours). The eye of the mollusk Bulla gouldiana expresses a circadian rhythm in optic nerve impulses that is generated by electrically coupled cells known as basal retinal neurons (BRNs). Daily fluctuations in the membrane potential of the BRNs appear to be driven by a rhythm in membrane conductance. Isolated BRNs exhibited spontaneous conductance changes similar to those observed in the intact retina. Membrane conductance was high in the late subjective night and decreased approximately twofold near projected dawn during at least two circadian cycles in culture. The persistence of daily conductance changes in isolated BRNs indicates that individual neurons can function as circadian pacemakers. PMID- 8421786 TI - Health research, a key to equity in health development. AB - This paper reinforces the idea of integrating more closely the social and medical sciences with an emphasis on people. The nexus between health and development is examined together with national research capability building as part of essential national health research progress. Suggestions are made identifying the most urgent requirements for further action. PMID- 8421787 TI - The promotion of health through planned sociopolitical change: challenges for research and policy. AB - The focus of health promotion is moving from the level of individuals to organizations, communities and broader social policy. Traditional quantitative methods (e.g. social surveys and experimental designs) which are variously appropriate at the level of individual behavior change require adaptation and refinement when sociopolitical change becomes the mechanism for health promotion. Because of their training and experience health services researchers and health educators (especially psychologists) are understandably resistant to necessary methodologic changes. Well designed and carefully conducted qualitative studies (e.g. ethnographic interviewing, participant observation, case studies and focus group activities) are required to complement quantitative approaches, and can fill gaps where quantitative techniques are suboptimal or even inappropriate: hard qualitative techniques can support soft quantitative methods. Their utility in process evaluation is now beyond dispute. Recent work at the New England Research Institute (NERI) is used to illustrate the role of qualitative research in the evaluation of health promotion through planned sociopolitical change. PMID- 8421788 TI - Geography of health: some trends and perspectives. AB - The present evolution of geography of health is highlighted through a selection of recent publications. Some current research trends are listed, such as disease ecology in a changing environment, geography of AIDS, urbanization, epidemiological transition, health care delivery. The most salient future perspectives are pointed out. PMID- 8421789 TI - Health transition: the cultural, social and behavioural determinants of health in the Third World. AB - The paper defines 'health transition' and outlines the development of recent research programmes. Evidence is reviewed as to the cultural, social and behavioural determinants of health in the Third World, and the extent to which they interact with the provision of health services in reducing mortality. Specific attention is given to the impact on mortality of education, and the historic experience of the now developed countries is compared with contemporary developing countries. Consideration is also given to the role of cultural factors and to radicalism, egalitarianism and the role of women in traditional society as well as fertility control and various forms of deleterious behaviour in contemporary society. The extent to which all these changes are facets of a single social transformation is discussed. Finally, the future of health transition research and its value for planned health interventions are summarized. PMID- 8421790 TI - Reflections on the economist and health economics in an international setting. PMID- 8421791 TI - In search of a contemporary theory for understanding mortality change. AB - This paper represents a speculative approach to the question of changing mortality levels in human populations. The surprising resilience and reluctance of mortality declines to respond to powerful countervailing is considered. A more integrated approach is proposed to examine the interactive roles of income, technology and behavior in relation to life expectancy. The paper concludes by a discussion of the relevance of the accumulation of health assets to national mortality levels. PMID- 8421792 TI - Health and poverty: past, present and prospects for the future. AB - Periodically the results of class comparisons in mortality rates have been reported. These reports have permitted comparisons since the earlier part of this century to the present period. The data thus available enables us to make some tentative predictions about the likely magnitude of class inequalities in mortality in the future. We consequently argue that: the concept of class should be abandoned in favour of a more direct measure of economic inequality which emphasises those living in poverty. despite overall declines in mortality for all socioeconomic groups, in the most recent period there has been an increase in the relative mortality disadvantage in some countries. this increase in mortality disadvantage is paralleled by an increase in the proportion of people, particularly children, living in poverty. Five groups constitute the bulk of those living in poverty and, of these, three (single mothers, the aged and the disabled) are likely to increase in numbers in the future, producing a likely increase in class-related mortality inequalities. Reducing these inequalities will depend upon welfare and education initiatives more than on any changes likely to be produced by the health system. PMID- 8421793 TI - Self, identity and the naming question: reflections on the language of disability. AB - With all the emphasis on 'political correctness', it is especially important to delineate the functions of naming. People with disabilities are facing issues quite similar to minority groups which have preceded them in attempting to enter 'mainstream' America. Their similarities and differences with these groups are traced as well as their own unique path (with all its implications) and some possible analytic and political solutions. PMID- 8421794 TI - Social research in health and the American sociopolitical context: the changing fortunes of medical sociology. AB - The fortunes of medical sociology, like other public policy-relevant disciplines, are shaped by political dynamics and prevailing values and attitudes. In the 1980s the field, which views disease and disability as consequences to a substantial degree of material conditions, social stratification, and inequalities among varying strata, lost ground to economics as societal attention focused on cost containment issues. Sociological concern with social structures clashed with dominant conservative and individualistic perspectives and the increased focus on personal responsibility and market strategies. There was decreasing tolerance in policy circles for the view that health, and the problems affecting disenfranchised groups such as the poor, the homeless, the uninsured and people with disabilities, were more due to our politics and social arrangements than the personal characteristics of those affected. Thus, little attention has been given in public health discourse to how life imperatives and social opportunities and constraints shape behavior. The paper documents the important role of the social sciences in health services research with special attention to examining the social context of the RAND Health Insurance Experiment. Although many of the questions medical sociology address sit on the periphery of policy-makers' concerns, a strong case is made for the revitalization of a critical scholarly role in medical sociology. I conclude, given the short and longterm problems we face in medicine and health, that we would have to invent a vigorous critical medical sociological enterprise if we did not already have one. PMID- 8421795 TI - An analysis of 9,918 consecutive perioperative autotransfusions. AB - The efficacy of perioperative autotransfusion (PAT) can be evaluated by the reduction in homologous transfusion accompanying its use. An alternative approach is to evaluate the amount of blood salvaged and retransfused. An analysis of 9,918 consecutive PAT procedures in various surgical specialties revealed that the average return of autologous blood salvaged was equivalent to 2.61 units of erythrocytes ("packed cells"). Cardiac operation had the greatest average number of units recovered (4.65), while orthopedic operation had the least (1.05). This method of analysis demonstrates that significant quantities of blood can be salvaged during PAT procedures. PMID- 8421796 TI - Modified Burch colposuspension for stress urinary incontinence in females. AB - Modified Burch colposuspension was performed upon 186 women having clinically and urodynamically verified stress urinary incontinence (SUI). After two years, 91 percent of the patients were cured or markedly improved. The success rate was as much as 95 percent among the patients who underwent Burch colposuspension as a first operative treatment for incontinence. In the patients with recurrent SUI, the cure rate was 82 percent. Failures appeared mostly during the first six months. There were no significant differences in cure rates between the patients who underwent concomitant hysterectomy and those who did not. The comparison of two different suture materials, Dexon (polyglycolic acid) and Maxon (polyglyconate monofilament absorbable) did not produce any significant differences. Temporary urinary retention was the most common postoperative complaint. During the six to 12 postoperative months, 19 percent of the patients experienced urgency and 9 percent had mild voiding difficulties. After one year, 12 percent of the patients had rectocele or enterocele, or both, which were surgically corrected. We conclude that modified Burch colposuspension is a safe and effective primary surgical method for the treatment of stress urinary incontinence in women. PMID- 8421797 TI - Duodenogastric reflux in patients with biliary lithiasis before and after cholecystectomy. AB - Duodenogastric reflux is quantified in ten patients with biliary lithiasis and a functioning gallbladder, before and six months after performing cholecystectomy. The results are compared with those of a control group (n = 10) with similar age and gender, without gastric or hepatobiliary pathologic factors. To evaluate reflux, we used six hour continuous intravenous infusion and subsequent determination in gastric juice of 99mTc-diethyliminodiacetic acid. Our results showed that patients with cholelithiasis have higher reflux rates than those in the control group (p < 0.001). When comparing patients before and after cholecystectomy, we confirmed that removal of the gallbladder produces a significant increase (p < 0.001) in reflux rates compared with preoperative values. PMID- 8421798 TI - Diminishing blood loss during operation for burns. AB - The current placebo-controlled, randomized clinical trial was done to determine the effect of preoperative 1-desamino-8-D-arginine vasopressin (DDAVP) infusion on blood loss in patients undergoing burn debridement and grafting, a patient population in which extreme blood loss is a frequent occurrence. Eleven patients undergoing 22 surgical procedures completed the study protocol--mean age was 33 years (range of 12 to 70 years), mean burn size was 53 percent body surface area (BSA) (range of 17 to 92 percent) and mean area debrided and grafted was 3,935 centimeters squared (range of 848 to 8,134) or 21.1 percent (range of 4.0 to 43.5 percent) BSA. The treatment group received 0.3 microliter per kilogram DDAVP infused during 15 to 30 minutes within one hour of anesthetic induction. The control group received placebo in a similar manner. Standard hemostatic maneuvers were used in all patients. Blood loss was calculated based on Warden's formula. No significant hemodynamic consequences or changes in routine coagulation profiles were noted in either group. No significant difference was found between the control and treatment groups in the volume of blood lost per percent BSA debrided and grafted (145.9 +/- 109.7 versus 130.2 +/- 61.7, respectively) or the volume lost per unit area debrided and grafted (0.75 +/- 0.54 versus 0.74 +/- 0.41, respectively). Based on these data, we cannot conclude that preoperative DDAVP infusion reduces blood loss in patients undergoing debridement and grafting of burn wounds. PMID- 8421799 TI - Wound healing after preoperative radiation for sarcoma of soft tissues. AB - Morbidity from wound healing was retrospectively analyzed in a series of 202 consecutive patients with tumors of the soft tissue of the extremities, torso and head and neck region who were treated with preoperative irradiation and conservative operation at the Massachusetts General Hospital between January 1971 and June 1989. A radiation boost dose was given to 143 patients (71 percent) postoperatively. The overall wound complication rate was 37 percent. One patient died because of necrotizing fasciitis. In 33 instances (16.5 percent), secondary operation was necessary, including six patients (3 percent) who required amputation. The wounds in the remaining 40 patients (20 percent) were treated without operation. Multivariate analyses of the data showed the factors that were significantly associated with wound morbidity: tumor in the lower extremity (p < 0.001), increasing age (p = 0.004) and postoperative boost with interstitial implant (p = 0.016). Accelerated fractionation (BID, two fractions per day) reached borderline statistical significance (p = 0.074). Two other factors showed association with wound morbidity by univariate analysis, but not in multivariate model: high pathologic grade (p = 0.02) and estimated volume of resected specimen > or = 200 milliliters (p = 0.065). Patient gender, intercurrent disease (diabetes or hypertension), obesity, maximal tumor size, primary versus recurrent tumor, duration of bed rest postoperatively, dose of postoperative boost radiation, the use of postoperative boost, the use of adjuvant chemotherapy and year of treatment did not show significant importance for wound morbidity. When the severe wound complications (defined as requiring secondary operation and including the patient who died because of necrotizing fasciitis) were considered, among all analyzed variables, only localization of tumor in the lower extremity as a single factor was significant (p < 0.001). Techniques for managing the wound are considered which are judged likely to contribute to a decrease of the incidence of wound healing delays. PMID- 8421800 TI - Total parathyroidectomy and presternal subcutaneous implantation of parathyroid tissue for renal hyperparathyroidism. AB - When hyperparathyroidism recurs in patients previously treated by total parathyroidectomy and intramuscular parathyroid autotransplantation, excision of the graft is not always technically easy and it is often necessary to resect the portion of the muscle containing the implants. During November 1986, we began a program to test the feasibility of presternal subcutaneous autotransplantation of parathyroid tissue. We hoped this technique would make removal of the grafts simpler. Thirty-six patients with renal hyperparathyroidism (RHPT) received subcutaneous parathyroid implants. Persistent or recurrent hyperparathyroidism was observed in three patients during the follow-up period. The presence of active parathyroid tissue was demonstrated after some time in all the patients. The parathyroid implants were easily excised in three instances (one persistent RHPT and two recurrences). Microscopic examination of the resected specimens did not show any sign of malignant transformation. We conclude that presternal subcutaneous implantation of parathyroid tissue after total parathyroidectomy is a quick, safe and efficient surgical procedure in the treatment of RHPT. PMID- 8421801 TI - Exulceratio simplex dieulafoy. AB - During a three year period, 18 patients with the Exulceratio Simplex Dieulafoy lesion of the stomach were treated (nine males and nine females). Because of intermittent bleeding episodes, lack of chronic ulceration of the gastric mucosa and the location of the lesion (most common in the upper part of the stomach), the lesion is easily overlooked at endoscopy and even at laparotomy with gastrotomy. Our treatment includes repeated endoscopies, attempted hemostasis with a combination of injection sclerotherapy and electrocoagulation and, if this is not successful, emergency surgical treatment with wedge resection using a technique with the TA 90 (U. S. Surgical Corp.) linear stapler. The diagnostic and therapeutic problems are discussed on the basis of patient profiles. PMID- 8421802 TI - Improvement of impaired pancreatic microcirculation by isovolemic hemodilution protects pancreatic morphology in acute biliary pancreatitis. AB - Impairment of the pancreatic microcirculation is a characteristic finding in experimental biliary pancreatitis. Isovolemic hemodilution with dextran 60 has been proven to maintain pancreatic capillary perfusion. To evaluate the significance of this therapeutic approach with respect to histologic changes, intravital microscopic assessment of the microcirculation was combined with a morphometric analysis of the pancreas by means of light microscopy in rabbits (n = 18). Pancreatic capillary perfusion was maintained in the rabbits subjected to hemodilution 30 minutes after the induction of pancreatitis with 54 percent of the capillaries still being perfused at 12 hours, compared with only 16 percent in the control group. The improved capillary perfusion resulted in a significant reduction of those changes considered potentially reversible (cell vacuolization and interstitial edema) that surround zones of necrosis. However, because of the early establishment of necrosis in this model, hemodilution was unsuccessful in preventing all cell death. Hemodilution can limit the progressive extension of pancreatic injury in this model of biliary pancreatitis. PMID- 8421803 TI - Limiting the anatomic extent of upper thoracic sympathectomy for primary palmar hyperhidrosis. AB - Ninety-four consecutive patients undergoing bilateral sympathectomy of the upper part of the thorax for primary palmar hyperhidrosis were reviewed. The supraclavicular operative approach was used and a limited sympathectomy was performed from below T1 to above T3, denervating the palm only. Follow-up evaluation was complete in 86 patients at a median period of 31 months. All patients had complete and permanent relief of palmar hyperhidrosis. However, 19 had compensatory hyperhidrosis and this was the common cause of patient dissatisfaction. Although axillary denervation was not performed, axillary sweating was a problem postoperatively in only two patients. Significant morbidity was minimal; the only permanent disability was in one patient with Horner's syndrome. Upper thoracic sympathectomy is a safe and effective method of treatment for primary palmar hyperhidrosis. The low incidence of compensatory sweating may be explained by the limited extent of the sympathectomy. Axillary sweating is rarely a significant postoperative problem, and extensive sympathectomy to include axillary denervation is unnecessary and should be avoided to minimize compensatory hyperhidrosis. PMID- 8421804 TI - Alterations in intracranial pressure and cerebral blood volume in endotoxemia. AB - Marked deterioration of neurologic function accompanies organ dysfunction in systemic sepsis. Although previous hypotheses have suggested that cerebral hypoperfusion, anoxia or progressive edema of the brain may be causative, the pathogenesis remains unknown. Patients with sepsis with stable or supported hemodynamics and adequate oxygenation may manifest confusion, stupor or coma. Recent evidence has demonstrated that the brain is the source of many classical mediators of inflammation after various forms of injury. These mediators, including the leukotrienes, have pronounced effect on cerebrovascular function. Endotoxin is known to stimulate the release of arachidonate from cell membranes, the rate limiting step in leukotriene synthesis. The current studies were performed to test the hypothesis that neurologic dysfunction associated with endotoxemia is characterized by alterations in cerebrovascular permeability or vasomotor function manifested by intracranial hypertension, or both. We studied the response of miniature swine to experimental endotoxemic shock and compared this response with hemorrhagic hypotension. We observed a dramatic elevation of intracranial pressure in swine subjected to endotoxemic shock, despite arterial hypotension. Moreover, estimation of cerebral blood volume (CBV) by reflectance infrared photoplethysmography demonstrated a dramatic increase in CBV, which corresponded to this elevation in intracranial pressure. However, cerebral cortical oxygen saturation was significantly reduced despite this net increase in CBV, indicative of an increase in the venous volume of the brain, while arterial volume remained the same or decreased from baseline levels. Oxygen extraction across the brain decreased during this same period compared with baseline and control values. These results demonstrate that endotoxemia is associated with the development of intracranial hypertension and an increase in CBV secondary to elevation of cerebrovascular venous volume coupled with reduced oxygen extraction across the brain. This evidence of cerebrovascular dysfunction probably represents blood flow maldistribution, similar to that seen in other organs with sepsis, suggesting a cause for altered neurologic function in systemic sepsis. PMID- 8421805 TI - Management of postoperative intra-abdominal abscesses by routine percutaneous drainage. AB - Postoperative intra-abdominal abscess (IAA) is a dreaded surgical complication. Percutaneous drainage (PD) has been offered as an alternative to surgical drainage for IAA because of the perceived lower morbidity and mortality rates. Seventeen consecutive unselected instances of IAA were reviewed to determine the value of the routine use of PD. Two patients with obvious anastomotic dehiscence and one with a retroperitoneal abscess were believed to be inappropriate for PD. Patients who could be managed by PD (group 1, n = 6) were similar to patients undergoing PD plus surgical drainage (group 2, n = 8) with respect to age, initial procedure, interval from operation to diagnosis of IAA and extent of organ failure. However, in group 2, more abscesses were greater than 5 centimeters (16.7 versus 62.5 percent), associated with polymicrobial growth (0.0 versus 50.0 percent), associated with enteric organisms (16.7 versus 100 percent; p < 0.05), accompanied by anastomotic dehiscence (16.7 versus 62.50 percent) and followed by a greater degree of mortality (16.7 versus 50 percent). Overall, only 33 percent of IAA were successful managed with PD alone. We conclude from these data that PD is of value only in selected instances of IAA. If an anastomotic dehiscence is a possibility, the patient should be managed operatively. PMID- 8421806 TI - Peripheral vascular injuries from plastic bullets in children. AB - During an eight month interval, 15 children and adolescents less than 20 years of age sustained vascular injuries to the lower extremities from plastic bullets. The plastic bullets had been introduced by the authorities as an attempt to reduce mortality from clashes between soldiers and demonstrators. Twenty-three vessels were injured in these 15 children: four patients had isolated arterial injuries, three had isolated venous injuries, and eight had combined injuries. The arterial injuries were repaired primarily or with reversed saphenous vein interposition grafts. The limb salvage rate was 100 percent, and there were no deaths. Eighty percent of the patients had palpable distal pulses observed at follow-up examination. We conclude from this experience that plastic bullets can cause serious injuries in children, vascular injuries from gunshots in children should be repaired or reconstructed using techniques perfected in the adult population, and low velocity gunshot vascular injuries in the pediatric age group can be successfully treated in a minimally equipped, developing world hospital by well-trained general surgeons. PMID- 8421807 TI - Less hepatic tissue necrosis after argon beam coagulation than after conventional electrocoagulation. PMID- 8421808 TI - Temporary end to side portacaval shunt in orthotopic hepatic transplantation in humans. PMID- 8421809 TI - The mythology of povidone-iodine and the development of self-sterilizing plastics. AB - The predominance of elemental iodine as a chemical antiseptic has been established during a century. Free iodine is effective for treatment and prevention of infection. Iodophors, such as povidone-iodine, have replaced elemental iodine in clinical use. Toxic absorption of povidone-iodine occurs from all tissues except intact adult skin, to which its use should be restricted. Povidone-iodine binds iodine so firmly that insufficient free iodine is released to be effective for treating or preventing infection. It is a weak antiseptic that is marginally acceptable as a disinfectant for adult skin. The shortcomings of povidone-iodine stimulated a search for iodophors that would liberate therapeutically effective concentrations of free iodine. These investigations led to a new self-sterilizing plastic formed by the complexing of polyurethane and iodine. PMID- 8421810 TI - Pathophysiologic factors and management of ascites. AB - Ascites is a manifestation of cirrhosis-induced portal hypertension. Pathogenesis is the result of a complex interaction of mechanical, humoral and neural events. Impaired excretion of sodium by the kidney is a hallmark of ascites, which is addressed by many of the available treatment options. Ascites contributes significantly to the morbidity and mortality rates of cirrhosis by increasing the likelihood of such fatal complications as variceal bleeding, hepatorenal syndrome and spontaneous bacterial peritonitis. Most ascitic patients respond to conservative or medical treatment. Five to 10 percent of the patients are refractory and may be candidates for surgical treatment. Peritoneovenous shunting is effective, and while safety is improved by following certain guidelines, its impact on survival is not clear. Portacaval shunting is also safe and effective. The use is accompanied by a prohibitively high morbidity rate because of encephalopathy, which limits its application despite what seems to be a significant impact on survival. PMID- 8421811 TI - [An intrauterine and prenatal fasciola hepatica infection in a dairy herd in South Netherlands]. AB - A intra-uterine infection with Fasciola hepatica in a three years old cow is proved on a milking cow herd in the South of the Netherlands. Two calves showed to have a prenatal infection. Further investigations on the farm revealed a general presence of this parasite. PMID- 8421812 TI - [A field study of the efficacy of ivermectin in propylene glycol in the treatment of mange in guinea pigs]. AB - The clinical manifestation of mange in a guinea-pig population in Utrecht is discussed. The etiology and the treatment with two subcutaneous injections of ivermectin 0.2 mg/kg with a ten-day interval, is described. A total of 37 guinea pigs with symptoms and 36 guinea-pig without signs of mange were treated. On the 21st day all previously infested guinea-pigs were free of symptoms. During this study side-effects occurred in 11 of the 73 animals. The most prominent side effects were a (sub-)dermal necrosis in the younger animals and an inflammation of the cutis in the older animals. The side-effects were probably caused by propylene glycol which was used as a diluent. PMID- 8421813 TI - [Could mycotoxins in Dutch pig-farming explain certain management problems?]. AB - The author reports a number of practical cases, indicating that moulds and mycotoxins may play an important role in Dutch pig farming. Therefore it seems desirable to him to form a study group that after investigation advises how problems with mycotoxins in farm animals can be restricted in the future. PMID- 8421814 TI - [Careful with trimethoprim in pregnant sows]. PMID- 8421815 TI - [Coughing adult cattle]. PMID- 8421816 TI - [Code for Competent Veterinary Practice]. PMID- 8421817 TI - Hyperfixation of HMPAO in subacute ischemic stroke leading to spuriously high estimates of cerebral blood flow by SPECT. PMID- 8421818 TI - Blood pressure after stroke. A one-year follow-up study. AB - BACKGROUND AND PURPOSE: Blood pressure changes in the year after acute stroke have been poorly documented. METHODS: We therefore studied blood pressure for 1 year after discharge from the hospital in 226 consecutive patients (mean age, 73 years) surviving an acute stroke. RESULTS: Marked increases (p < 0.001) in mean systolic and mean diastolic blood pressures were seen in two thirds (69%) of the patients 1 month after discharge, and blood pressure remained stable at this level during the remainder of the follow-up year. Similar blood pressure changes were seen irrespective of sex, final stroke diagnosis, or whether the patient had a history of hypertension before the stroke. Patients with a history of hypertension had significantly higher blood pressures (p < 0.001) throughout the follow-up year than previously normotensive patients. One month after discharge blood pressure was found to have decreased in 31% of the patients; these were older and had a higher mortality during the follow-up year than patients with blood pressure increases. About 20% of all patients suffered from orthostatism (defined as a decrease in systolic blood pressure of > or = 20 mm Hg when rising from the supine position to standing). CONCLUSIONS: We conclude that antihypertensive treatment should not be reduced before discharge from the hospital and that blood pressure should be checked about 1 month after discharge. We suggest that standing blood pressure also be measured to make an appropriate treatment decision. PMID- 8421819 TI - Recovery of functional status after stroke. A postrehabilitation follow-up study. AB - BACKGROUND AND PURPOSE: Information on predictors of long-term change in functional capacity after a rehabilitation program in stroke patients is scant. This study describes the long-term evolution of self-reported functional ability after discharge from rehabilitation and its relation with age, level of neural impairment at discharge, and changes in neural impairment during follow-up. METHODS: Fifty patients (31 men and 19 women; mean +/- SD age, 66.0 +/- 9.9 years; range, 47-86 years) with a first unilateral stroke and no severe cognitive impairment were consecutively enrolled. Self-reported disability in activities of daily living and neural impairment measured by the Fugl-Meyer Scale were evaluated after discharge from a rehabilitation program and 3 and 6 months later. RESULTS: Functional disability was significantly reduced after 3 and 6 months. Attenuation of disability occurred mainly among those patients with more severe baseline neural impairment. In this group, patients aged > or = 65 years were more disabled at baseline than younger individuals, but they had the same rate of improvement. In patients aged < 65 years, changes in disability over time could be attributed to changes in neural function, whereas older patients' functional recovery was greater than that expected from their improvement in neural impairment alone. CONCLUSIONS: These results suggest that in stroke patients with severe neural damage further functional improvement occurs even after completion of a rehabilitation program. There is evidence that older patients may be more likely to employ compensatory strategies to overcome some of the neural impairment that remains after stroke. PMID- 8421820 TI - Distribution of atherosclerosis and risk factors in atherothrombotic occlusion. AB - BACKGROUND AND PURPOSE: The present study was performed to determine the relation between distribution of atherosclerosis and risk factors in Japanese patients with atherothrombotic occlusion. METHODS: We studied 154 patients with atherothrombotic occlusion of the extracranial internal carotid artery (n = 75, ICA group), the horizontal portion of the middle cerebral artery (n = 47, MCA group), and the basilar artery (n = 32, BA group), all of which were confirmed by cerebral angiography. We investigated the distribution of atherosclerosis of the three sites and compared the risk factors for atherosclerosis between the three groups. We used 113 subjects without stroke as the control group. RESULTS: A strong correlation was present between atherosclerosis of the extracranial internal carotid artery and that of the basilar artery. However, only a weak correlation existed between atherosclerosis of the middle cerebral artery and that of the other vessels. Although the prevalence of smoking and hypertension was higher and high density lipoprotein cholesterol levels were lower in the three groups than in the control subjects, no significant differences were found in age, sex, prevalence of smoking and hypertension, serum levels of triglycerides, and high density lipoprotein cholesterol among the three groups. The prevalence of coronary heart disease and diabetes mellitus and the serum levels of hemoglobin A1c, total cholesterol, and low density lipoprotein cholesterol were higher in the ICA and BA groups than in the MCA group. The prevalence of electrocardiographic evidence of left ventricular hypertrophy was higher in the MCA group than in the other groups. CONCLUSIONS: In addition to smoking, hypertension, and low concentration of high density lipoprotein cholesterol, diabetes mellitus and hypercholesterolemia seem to be associated with atherosclerosis of the extracranial internal carotid artery and the basilar artery, and advanced hypertension may play a role in the development of middle cerebral artery occlusion. PMID- 8421821 TI - Secular trend of mortality from cerebral infarction and cerebral hemorrhage in Taiwan, 1974-1988. AB - BACKGROUND AND PURPOSE: Pathological patterns of stroke are different in various races, with the predominant stroke type in the Chinese being intracerebral hemorrhage. A total of 31,078 deaths from cerebral infarction and 77,773 deaths from cerebral hemorrhage in Taiwan were reported for groups of subjects aged 40 79 years during the period 1974-1988 to elucidate their secular trends. METHODS: Vital statistics and demographic data were collected for analyzing the truncated age-adjusted mortality rates. Poisson regression analysis was used to estimate the relative risk and 95% confidence interval. RESULTS: The mortality rates for cerebral hemorrhage were greater than those for cerebral infarction. The mortality rates increased exponentially with age for both subtypes. The decline in age-specific mortality for the period 1974-1983 was much more striking for cerebral hemorrhage than for cerebral infarction. Mortality from cerebral infarction and hemorrhage increased with age for all birth cohorts except the cerebral hemorrhage mortality of the oldest cohort. Male/female ratios for both cerebral infarction and hemorrhage were greater in the younger age groups. The cerebral hemorrhage/infarction ratio during the period 1984-1988 was highest for the younger age groups and lowest for the oldest age groups. CONCLUSIONS: The different secular trends of mortality from cerebral infarction and hemorrhage imply that these two patterns of stroke may be associated with some different risk factors. PMID- 8421822 TI - Increased thromboxane biosynthesis in patients with acute cerebral ischemia. AB - BACKGROUND AND PURPOSE: Clinical and experimental studies suggest that platelets have a major role in the pathogenesis of cerebral ischemia. However, ex vivo both platelet aggregation studies and measurements of platelet-derived products in patients with cerebral ischemia have shown inconsistent results. The present study was designed to resolve this inconsistency. METHODS: We have measured the urinary excretion of a thromboxane metabolite, 11-dehydro-thromboxane B2, by a previously validated radioimmunoassay technique in 51 patients with acute cerebral ischemia who had experienced either a transient ischemic attack (14 patients) or an ischemic stroke (37 patients) and in 20 control patients with nonvascular neurological disorders. The median time between the onset of symptoms and urine sampling was 24 hours (range, from 2 hours to 8 days). RESULTS: The excretion rate of immunoreactive 11-dehydro-thromboxane B2 ranged between 39 and 478 pmol/mmol creatinine in patients with a transient ischemic attack and between 23 and 5,916 pmol/mmol creatinine in stroke patients, with 29% (p = 0.18) and 51% (p = 0.004) of the urine samples, respectively, exceeding the upper limit of the control samples (251 pmol/mmol creatinine [mean +/- 2 SD]) (p = 0.01). In stroke patients, metabolite excretion was not related to the type (cortical or "lacunar") or site of cerebral infarction. Low-dose aspirin (50 mg per day for 7 days) reduced the urinary excretion by approximately 85% in 11 consecutive stroke patients. CONCLUSIONS: We conclude that 1) episodes of enhanced thromboxane biosynthesis are detected infrequently in patients with a transient ischemic attack, 2) aspirin-suppressible episodes of increased thromboxane formation can be detected during the early phase of acute ischemic stroke, and 3) this finding may provide a rationale for testing the efficacy and safety of this drug in this setting. PMID- 8421823 TI - Free protein S deficiency in acute ischemic stroke. A case-control study. AB - BACKGROUND AND PURPOSE: Deficiency of free protein S, a naturally occurring anticoagulant, may be acquired in the setting of acute illness and increasingly has become recognized as a possible stroke risk factor. We sought to determine whether free protein S deficiency is associated with acute cerebral infarction among older individuals at risk for stroke. METHODS: Free protein S was measured by Laurell rocket immunoelectrophoresis in 94 adults admitted for acute cerebral infarction and in 94 hospitalized control subjects of similar age, sex, and race. Patients with a history of cerebrovascular disease, acute thrombotic or hematologic diseases, or medical conditions known to cause free protein S deficiency were excluded from the control group. RESULTS: The percentage of patients with free protein S deficiency (< 20% normal total protein S) was similar in the case and control groups (21% versus 20%, respectively). Among all subjects, free protein S deficiency was more common in blacks than nonblacks (34% versus 13%, p = 0.001). A very low free protein S (< 15% normal total protein S) was more frequent among case patients than control subjects (11% versus 5%), but this trend failed to reach statistical significance. CONCLUSIONS: Free protein S deficiency is common among hospitalized patients, even in the absence of a recognized predisposing condition. Our findings indicate that acquired deficiency of free protein S is not a major risk factor for ischemic stroke. PMID- 8421824 TI - Differentiation of multi-infarct and Alzheimer dementia by intracranial hemodynamic parameters. AB - BACKGROUND AND PURPOSE: The differentiation between the Alzheimer and multi infarct types of dementia may still be equivocal considering clinical criteria, neuropsychological tests, and imaging techniques. Cerebral microangiopathic alterations underlying multi-infarct dementia should allow the characterization of dementia subgroups. METHODS: Patients with a diagnosis of multi-infarct dementia (n = 17; mean age, 69.1 +/- 8.5 years) or Alzheimer dementia (n = 24, mean age, 65.8 +/- 9.0 years) according to standard testing criteria, clinical findings, and neuroimaging techniques (computed tomography and magnetic resonance imaging) were investigated prospectively by transcranial Doppler sonography and compared with a normal reference group (n = 64; mean age, 61.0 +/- 11.1 years). Transcranial Doppler sonography allows an indirect evaluation of peripheral flow resistance in the microcirculatory bed by quantifying pulsatility characteristics, as reflected in the effective pulsatility range (time-averaged mean blood flow velocity minus the peak-systolic to end-diastolic amplitude, in centimeters per second). RESULTS: A total of 204 vessels were investigated in 105 subjects. Mean and diastolic blood flow velocities as well as the effective pulsatility range were significantly lower in the multi-infarct dementia group compared with the Alzheimer and the normal reference groups (p < 0.001). By using receiver operating characteristic analysis, a cutoff point for effective pulsatility range values of -5 cm/sec gives a side-dependent sensitivity of 90.48 95.24% and a specificity of 64.71-70.59% in diagnosing Alzheimer-type dementia; the corresponding sensitivity and specificity for a value of -2 cm/sec are 82.35 88.24% and 80.95-90.48%, respectively. CONCLUSIONS: Pulsatility changes as reflected by the effective pulsatility range are a noninvasive additional criterion in the differential diagnosis of dementia. PMID- 8421825 TI - Leukocyte infiltration in acute hemispheric ischemic stroke. AB - BACKGROUND AND PURPOSE: The dynamics of leukocyte infiltration in human cerebral ischemia were studied using technetium-99m hexamethylpropyleneamine oxime (99mTc HMPAO)-labeled leukocyte brain single-photon emission computed tomography (SPECT). METHODS: Twenty-two patients diagnosed as having hemispheric ischemic stroke were examined with 99mTc HMPAO brain SPECT for cerebral blood flow study and 99mTc HMPAO-labeled leukocyte brain SPECT for the study of leukocyte infiltration. Three patients with chronic hemispheric ischemic stroke received one examination. Nineteen patients with acute hemispheric ischemic stroke received their initial examination within the first after onset. Follow-up examinations were performed at intervals of 1-3 weeks whenever possible. RESULTS: In patients with chronic hemispheric ischemic stroke, leukocyte infiltration was not seen in areas of perfusion defect. In patients with acute hemispheric ischemic stroke, leukocyte infiltration was seen in areas of perfusion defect during the acute stage, which persisted for no less than 5 weeks after onset and then declined. CONCLUSIONS: A new method to study and monitor the process of leukocyte infiltration in acute cerebral ischemia using 99mTc HMPAO-labeled leukocyte brain SPECT is described. This method shows that leukocyte infiltration in acute hemispheric ischemic stroke is a dynamic process that persists for no less than 5 weeks and then declines. PMID- 8421826 TI - Age as a modifying factor on the effect of antihypertensive therapy in focal stroke in rats. AB - BACKGROUND AND PURPOSE: Antihypertensive treatment with hydralazine for 10 weeks but not 6 weeks reduces infarct size in 13-week-old spontaneously hypertensive rats subjected to focal cerebral ischemia. This study was designed to examine whether the duration of treatment needed to reduce infarct size depends on how long hypertension is present before the initiation of antihypertensive therapy. METHODS: Six-week-old spontaneously hypertensive rats were treated for 6 weeks and 10-month-old spontaneously hypertensive rats for 10 weeks with 20 mg/kg hydralazine added daily to the drinking water. The animals were then subjected to focal cerebral ischemia by tandem permanent common carotid and middle cerebral artery occlusion. RESULTS: Blood pressure in the treated groups was lower than that in the untreated groups for the entire treatment period in both experiments. Infarct volume in 10-month-old spontaneously hypertensive rats treated for 10 weeks, but not in 6-week-old spontaneously hypertensive rats treated for 6 weeks, was significantly less than in untreated controls (p = 0.02). CONCLUSIONS: This study emphasizes the importance of duration of antihypertensive treatment in reducing infarct volume in spontaneously hypertensive rats after focal cerebral ischemia and demonstrates that the effect appears to be independent of the duration of hypertension before the initiation of treatment. PMID- 8421827 TI - Prophylactic effect of imidapril on stroke in stroke-prone spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: It has been reported that some angiotensin converting enzyme inhibitors can prevent stroke-prone spontaneously hypertensive rats from stroke at much higher doses than clinical doses used for hypertension therapy. This study was performed to investigate the prophylactic effectiveness of imidapril against stroke in comparison with enalapril. METHODS: Salt-loaded stroke-prone spontaneously hypertensive rats were orally given imidapril (0.5, 1, 2, and 5 mg/kg per day), enalapril (2 and 5 mg/kg per day), or hydralazine (5 mg/kg per day). Stroke signs were scored, and blood pressure, protein concentration, and N-acetyl-beta-D-glucosaminidase activity in urine were measured. After 2 weeks of medication, angiotensin converting enzyme activities in the aorta were measured 24 hours after dosing. RESULTS: In the control group, severe hypertension developed, and all rats died within 12 weeks because of stroke. Imidapril and enalapril dose-dependently decreased the stroke-related mortality, and both agents at 5 mg/kg per day showed excellent prophylaxis, although they did not inhibit hypertensive development. Imidapril at 0.5 mg/kg per day significantly prevented stroke to almost the same extent as enalapril at 2 mg/kg per day or hydralazine at 5 mg/kg per day. Imidapril dose-dependently suppressed the elevation of the two urinary indexes, which was followed by stroke. Imidapril inhibited enzyme activity in the aorta more strongly than did enalapril at the same dose. CONCLUSIONS: Imidapril prevented the incidence of stroke in stroke-prone spontaneously hypertensive rats at a dose of 0.5 mg/kg per day or more by amelioration of kidney dysfunction. Reduction of blood pressure is not necessary, although enzyme inhibition in the vasculature may partly relate to the effect. PMID- 8421828 TI - Spatial features of focal infarction after hydralazine treatment in stroke-prone spontaneously hypertensive rats. AB - BACKGROUND AND PURPOSE: Rapid occlusion of the middle cerebral artery above the rhinal fissure produces a large ischemic infarct in hypertensive rats, but this occlusion results in a minimal lesion in young normotensive rats. Our purpose was to attenuate rising blood pressure in young stroke-prone spontaneously hypertensive rats with hydralazine before the occlusion to determine if the gross infarct volume is smaller, and if it is, to determine whether length, width, depth, or surface area of the infarct changes, which could suggest a mechanism of protection. METHODS: Untreated rats (n = 6) and rats receiving hydralazine for 1.5 (n = 6) or 5 (n = 5) weeks were anesthetized, and the middle cerebral artery was rapidly occluded with a ligature. One day later the rats were killed and the brains were fixed in formalin. Fine-grain-release film that is sensitive to spectral properties of the infarct was used to photograph the tissue. Infarcted areas were traced on paper and then digitized for measurements and computations with a microcomputer. RESULTS: Compared with untreated rats, tail systolic blood pressure (120 +/- 3 versus 138 +/- 4 mm Hg), infarct volume (61 +/- 4 versus 93 +/- 6 mm3), infarct surface area (39 +/- 1 versus 54 +/- 2 mm2), infarct width (3.8 +/- 0.1 versus 4.8 +/- 0.2 mm), and infarct length (6.0 +/- 0.3 versus 8.1 +/- 0.3 mm) were less in rats receiving hydralazine for 5 weeks (p < 0.05). No change was detected in infarct depth. CONCLUSIONS: Treatment of young stroke prone spontaneously hypertensive rats with hydralazine for 5 weeks before middle cerebral artery occlusion results in a smaller infarct. The narrower, shorter dimensions indicate increased protection against lateral enlargement of the infarct and the possibility that protection was due to increased collateral blood flow through modified blood vessels. PMID- 8421829 TI - A new approach to the integrity of dual blood-brain barrier functions of global ischemic rats. Barrier and carrier functions. AB - BACKGROUND AND PURPOSE: We studied the influence of reperfusion on carrier and barrier functions of the blood-brain barrier after transient global ischemia in rats. METHODS: We used iodine-125-labeled 3-iodo-alpha-methyl-L-tyrosine and carbon-14-labeled sucrose as tracers for studying carrier and barrier functions of the blood-brain barrier, respectively. Brain uptakes of these two tracers were measured in Wistar rats subjected to either 15- or 30-minute four-vessel occlusion developed by Pulsinelli and Brierly before recirculation for 3, 6, 24, 48, and 72 hours. Tracer (5 microCi) was injected intravenously in each rat 30 minutes before killing the animal. RESULTS: Following 15- or 30-minute ischemia, [14C]sucrose uptakes were significantly higher at 3 and 6 hours of reperfusion before recovery to control values after reperfusing for 24 to 48 hours in almost all brain regions. However, a rebound in radioligand uptake was significantly manifested in some sites at 72 hours after reperfusion (p < 0.05 to p < 0.01). Uptakes of 125I-3-iodo-alpha-methyl-L-tyrosine were brain site-dependent: significantly (p < 0.05) higher in cortex (3 and 48 hours after reperfusion) and thalamus (3, 6, and 48 hours after reperfusion) but significantly (p < 0.05 to p < 0.01) lower in striatum, cortex (72 hours after reperfusion), and midbrain (6, 24, and 72 hours after reperfusion). Because the [14C]sucrose uptake in brain was 10% lower than that of 125I-3-iodo-alpha-methyl-L-tyrosine, the change in absolute transport of the latter tracer was approximated to its brain uptake. CONCLUSIONS: The carrier and barrier functions of the blood-brain barrier should be evaluated separately. The radioligand 125I-3-iodo-alpha-methyl-L-tyrosine may serve as a useful tool to evaluate the carrier function of the blood-brain barrier after transient cerebral ischemia in rats. PMID- 8421830 TI - Comparative histopathologic consequences of photothrombotic occlusion of the distal middle cerebral artery in Sprague-Dawley and Wistar rats. AB - BACKGROUND AND PURPOSE: We have developed a minimally invasive model of photothrombotic occlusion of the distal middle cerebral artery in rats and have evaluated the patterns and features of the resulting histopathologic injury in two normotensive strains. METHODS: Food-deprived male Sprague-Dawley (n = 14) and Wistar (n = 10) rats anesthetized with halothane/nitrous oxide underwent a small craniotomy to expose the right distal middle cerebral artery just above the rhinal fissure. The animals were injected intravenously with the photosensitizing dye rose bengal, and the distal middle cerebral artery was irradiated with light from an argon laser-activated dye laser at three separate points to induce thrombotic occlusion. The ipsilateral common carotid artery was then permanently occluded, and the contralateral common carotid artery was occluded for 60 minutes. Three days later, the brains were perfusion-fixed and prepared for histopathologic examination, and infarct volume was determined by quantitative planimetry. RESULTS: In Sprague-Dawley rats, a large consistent temporoparietal cortical infarct was observed; mean +/- SD infarct volume was 130.5 +/- 40.0 mm3 (coefficient of variation, 30.7%) and a relatively small adjacent zone of selective neuronal necrosis ("incomplete infarction"), amounting to only 9.1% of the total injury volume, was also seen. By contrast, Wistar rats had smaller and more variable cortical infarcts (volume, 48.4 +/- 26.9 mm3; coefficient of variation, 55.6%) but displayed a much more substantial zone of incomplete cortical infarction (volume, 20.8 +/- 10.1 mm3; 30.1% of the total injury volume). In neither strain was infarct size related to alterations of blood pressure. In both strains, infarcts were limited to the cortex, typically involving the parietal cortex, somatosensory cortex, and forelimb region. Three rats exhibited infarcts in the contralateral hemisphere. CONCLUSIONS: This model has the advantages of necessitating only minimal surgery, allowing the dura to remain intact, and avoiding mechanical trauma to the brain surface. In Sprague Dawley rats, the resulting large cortical infarct exhibited relatively small interanimal variation, making the model suitable, for example, for replicate studies of pharmacotherapy. In Wistar rats, the large zone of incomplete infarction, a unique feature heretofore undescribed in rodent models of permanent focal ischemia, lends the model to the study of the pathomechanisms underlying graded cortical ischemic injury. PMID- 8421831 TI - Middle cerebral artery occlusion without craniectomy in rats. Which method works best? AB - BACKGROUND AND PURPOSE: Our purpose was to assess the effectiveness of middle cerebral artery occlusion in producing acute focal ischemia in the rat by the use of Koizumi's and Longa's methods, in which occlusion is achieved by passing a nylon thread into the internal carotid artery. METHODS: Cerebral blood flow was measured by using the hydrogen clearance method, and the brains were examined histologically to assess ischemic damage. RESULTS: By Koizumi's method profound reduction in cerebral blood flow was achieved in 28 of 30 rats (93%). The mean cerebral blood flow in the middle cerebral artery territory was 10.7 (95% confidence interval, 9.9-11.5) ml/100 g per minute. By Longa's method reduction in cerebral blood flow was achieved in only 29 of 52 rats (56%), and in these animals mean cerebral blood flow was 33 (95% confidence interval, 28.3-33.7) ml/100 g per minute (p < 0.001 compared with Koizumi's method). Cerebral blood flow was reduced to < 16 ml/100 g per minute in only seven animals (24%). CONCLUSIONS: By Koizumi's method the depth of ischemia is more profound, occlusion is achieved in a much higher proportion of cases, and the incidence of perforation of the intracranial internal carotid is much less frequent than by Longa's method. PMID- 8421832 TI - Role of transcranial Doppler in neuroradiological treatment of intracranial vasospasm. AB - BACKGROUND AND PURPOSE: The expanded role of interventional neuroradiological treatment for post-subarachnoid hemorrhage vasospasm has highlighted the diagnostic utility of transcranial Doppler studies in this condition. The role of transcranial Doppler in follow-up and determining the need for repeat intervention has not been previously emphasized. SUMMARY OF REPORT: Intracranial angioplasty for clinically evident vasospasm after subarachnoid hemorrhage was performed in four patients. In two patients, transcranial Doppler flow velocities remained elevated despite initial anatomic correction of the vasospasm. Reangiography revealed new areas of involvement by vasospasm. Reangioplasty or papaverine infusion treatment of the new lesions resulted in decreased flow velocities and clinical improvement in all patients. CONCLUSIONS: Transcranial Doppler has a more significant role than has been previously emphasized in the management of patients undergoing interventional neuroradiological treatment for intracranial vasospasm. Specifically, the persistence of elevated transcranial Doppler flow velocities after intracranial angioplasty suggests the need for repeat angiographic evaluation and possibly further therapy. PMID- 8421833 TI - Cerebral blood flow in mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes. AB - BACKGROUND AND PURPOSE: The precise mechanism of neurological symptoms with mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes (MELAS) is still controversial. We investigated the correlation between strokelike episodes and cerebral blood flow in two patients with MELAS and discuss the pathogenesis of strokelike episodes with MELAS. SUMMARY OF REPORT: Cerebral dynamic computed tomography and cerebral angiography were used to measure cerebral circulation in the first case, that of a 20-year-old woman with MELAS. The second subject was a 13-year-old female who was studied with xenon enhanced computed tomography. The cerebral blood flow studies were performed 3-72 hours after the onset of strokelike episodes. Serial cerebral angiography, dynamic computed tomography, and xenon-enhanced computed tomography showed vasodilation localized in the affected cerebral cortexes during strokelike episodes, without any reduction in regional cerebral blood flow. CONCLUSIONS: Our study suggests that the strokelike episodes associated with MELAS are different in origin from ischemic stroke. PMID- 8421834 TI - Possible association of ischemic stroke with phentermine. AB - BACKGROUND AND PURPOSE: Some commonly used anorexiants, including methamphetamine and phenylpropanolamine, have been associated with stroke. Because phentermine is an anorexiant with a chemical structure similar to that of amphetamines, similar side effects might be expected. CASE DESCRIPTIONS: Two patients using phentermine (one was also using phendimetrazine) developed ischemic cerebrovascular disease. One suffered a cerebral infarct with angiographic evidence of vasculopathy involving multiple vascular beds. The other patient developed headache and a hemisensory disturbance of 7 days' duration. CONCLUSIONS: Phentermine, and possibly phendimetrazine, should be considered an anorexiant and sympathomimetic drug that can be associated with ischemic cerebrovascular disease. PMID- 8421835 TI - Carotid artery intraplaque hemorrhage and stenotic velocity. AB - BACKGROUND AND PURPOSE: One of the proposed mechanisms for sudden expansion of a carotid bifurcation plaque is hemorrhage within the lesion. It has been postulated that the sudden increase in plaque size will acutely reduce blood flow to the ipsilateral hemisphere and induce either a transient ischemic attack or a stroke. In this study, the relation between peak systolic velocity at the site of narrowing and its potential role in the development of intraplaque hemorrhage were investigated. METHODS: Ten patients who had carotid endarterectomy were examined by duplex Doppler sonography before surgery to determine the peak systolic velocity at the site of maximal narrowing. The excised carotid plaques were sectioned at 1-mm intervals and examined for histological evidence of intraplaque hemorrhage. The recorded peak systolic velocities in patients with intraplaque hemorrhage were compared with the velocities in cases in which no hemorrhage was identified. RESULTS: Five of the ten patients had intraplaque hemorrhage. Four of the five patients with intraplaque hemorrhage had a peak systolic velocity of > 420 cm/sec and diastolic velocities of > 160 cm/sec; none of the patients without intraplaque hemorrhage had such high values. CONCLUSIONS: Peak systolic velocity is significantly higher in patients with intraplaque hemorrhage. The specificity and sensitivity of a peak systolic velocity of > 420 cm/sec in predicting intraplaque hemorrhage remains to be determined. PMID- 8421836 TI - Measuring quality of life in stroke. AB - BACKGROUND AND PURPOSE: Little attention has been focused on quality of life in stroke outcome research. The purpose of this review is to outline the meaning of the concept, describe important methodological issues and methods of assessment, review existing quality of life measures, and discuss criteria for selecting an appropriate instrument. SUMMARY OF REVIEW: The following 10 quality of life instruments were reviewed: COOP Charts; Euroqol; Frenchay Activities Index; Karnofsky Performance Status Scale; McMaster Health Index Questionnaire; Medical Outcomes Study 20-Item Short-Form Health Survey; Nottingham Health Profile; Quality of Life Index; Quality of Well-being Scale; and the Sickness Impact Profile. They were evaluated in terms of length, time needed to complete, content, scoring, and psychometric characteristics. CONCLUSIONS: Emphasis should be placed on further psychometric evaluation of existing quality of life measures rather than on generating new instruments. There is particular need for supplementary data on the responsiveness of the instruments to changes in patients' clinical status over time. The choice of a suitable quality of life instrument should be based not only on psychometric properties but also on careful consideration of the research question, the relevance to the objectives of the study, the feasibility of the instrument, and the specific characteristics of the stroke patients under investigation. PMID- 8421837 TI - Small, deep, penetrating cerebral artery embolic cerebral infarct or embolic lacune? PMID- 8421838 TI - Monkey model of complete global ischemia. PMID- 8421839 TI - [Neonatology yesterday, today, tomorrow]. PMID- 8421840 TI - [Description of activities in the neonatal department at Rigshospitalet during the period 1983-1990]. AB - Since 1983, the gestational age (GA in completed weeks), use of mechanical ventilation, duration of hospitalization and the diagnosis-related group were registered routinely for all infants admitted to the Neonatal Department, Rigshospitalet, Copenhagen. The data were analysed concerning the 6,636 infants treated during the seven-year period until the end of 1991. The annual number of admissions decreased during the period (p < 0.001) whereas the number of extremely preterm infants (GA < 28 weeks) increased from 30 to 45 annually (p < 0.05). The total mortality remained constant at about 7% of all admissions but decreased significantly for the extremely preterm infants. The use of mechanical ventilation was almost halved during the period: from 235 to 146 annually. Even where the extremely preterm infants were concerned, the use of mechanical ventilation decreased from 77% to 51% (p < 0.05). The gestational age specific duration of hospitalization for the surviving infants remained constant. The number of infants with surgical anomalies or congenital heart disease increased among the infants born at term. The cost of treatment increased by 40% in fixed prices to DDK 4,000 per day (approximately 333 pounds). The authors conclude that the introduction of nasal continuous positive airway pressure (CPAP) has permitted treatment of moderately preterm infants in county hospitals and has also resulted in avoidance of mechanical ventilation in extremely preterm infants without preventing improved survival. Although the patients have become more selected, the duration of hospitalization remains unchanged. PMID- 8421841 TI - [Structural and functional cerebral changes in patients with schizophrenia]. AB - During the past decades the possibility of measuring regional brain structure and function by computed tomography, magnetic resonance, single photon emission computed tomography and positron emission tomography has brought the biological research on schizophrenia into focus. Structural changes with enlargement of the lateral and third ventricle and broader sulci on the brain surface compared to those of healthy volunteers have been reproducible findings. Functionally, the reduced activity in the prefrontal cortex especially during activation tests has been the most reliable finding. However, the etiology and specific meaning of the brain changes remains unsolved. Furthermore, in all the studies performed there is a considerable overlap in the measurements in the schizophrenic patients and in the controls. PMID- 8421842 TI - [Transport of sick newborn infants]. PMID- 8421843 TI - [Juvenile recurrent parotitis]. AB - Juvenile recurring parotitis is a relatively rare condition. The clinical picture as illustrated by a characteristic case history is reviewed. The condition shows a tendency to remission around puberty. Sialographic examination is characterised by the configuration of the parotid gland said to resemble a bunch of grapes. Biochemical tests do not contribute to the diagnosis which is established by the clinical picture and the typical sialographic findings. No systemic disease can be demonstrated in the patients who are healthy in other respects. The cause of the condition appears to be congenital sialectic changes localised to the parotid gland with resultant secondary ascending bacterial infection. In treatment during the acute stages, attempts should be made to stimulate secretion of saliva with sour foodstuffs and chewing gum. Treatment with penicillin for one week is recommended. On account of the benign character of the condition and the good long-term prognosis, operative intervention should not prove necessary. PMID- 8421844 TI - [Deficiency of mannan-binding protein--a recently discovered complement defect syndrome]. AB - Mannan-binding protein (MBP) is a newly discovered serum protein which activates the complement system by the classical pathway by binding to mannose-rich surfaces on microorganisms. MBP deficiency predisposes to infection in infancy. MBP is a C-type lectin with collagen structure, a so-called collection. The paper discusses the structure and physiological functions of MBP. MBP may be an important parameter in the investigation of immunodeficiencies in children. PMID- 8421845 TI - [A stink bomb in an office environment. Sick building syndrome with toxic rhinitis after exposure to fusel]. AB - In 1983 WHO, defined "The Sick Building Syndrome". Various conditions influence the indoor climate, among others the degassing of chemicals. An epidemic of sick building syndrome is described in a two year old office building. The symptoms appeared after exposure to toxic chemicals released by a stink bomb--a form of exposure not previously described in the literature. Gas chromatographic analysis of the content of the stink bomb revealed 22 different chemicals likely to be remains from an alcoholic fermentation process. Twenty-four employees were exposed. A questionnaire investigation of the employees revealed that seven had symptoms related to the exposure. A clinical investigation of those who claimed to have symptoms took place. Six of the seven patients were investigated. They all had toxic rhinitis with bleeding. Owing to an unsystematic procedure it took more than two months before the indoor climate was normalized. The toxic rhinitis and other symptoms gradually decreased over more than four months. In order to minimize potential health damage due to the sick building syndrome, we recommend that experts should be consulted within this particular field. PMID- 8421846 TI - [Should femoral neck fractures be treated as emergency surgery?]. PMID- 8421848 TI - [Aspiration of a coin after correct use of Rotahaler with Ventoline inhalation powder]. AB - Rotahalers are widely used to administer Ventoline and Becotide. We report a case of inhalation of a coin from a rotahaler in a 71 year old asthmatic male and discuss prophylaxis. PMID- 8421847 TI - [Electrocardiography and serum CK-MB determination in myocardial infarction and reperfusion in patients treated with intravenous streptokinase or placebo]. PMID- 8421849 TI - [Risks for kidney donors]. PMID- 8421850 TI - [A broken hip]. PMID- 8421851 TI - [Accidental falls of the elderly]. PMID- 8421852 TI - [Transportation of sick newborn infants]. AB - In a six month period, 96 newborn infants were transferred from local hospitals to the Department of Neonatology, State University Hospital (Rigshospitalet), Copenhagen. Complications related to transport and potential risk factors were registered prospectively. Considering the condition of the infants, 31% of the transports were not carried out under optimal conditions. Serious complications occurred in 13% of the transports of infants at risk for cardio-pulmonary insufficiency. In 25% of the intubated infants, complications related to displacement of the tracheal tube were observed on arrival, but in only one of these cases the complication was recognized and treated during transport. Hypothermia occurred frequently in low birth weight infants. Failing monitoring equipment and difficulties with stethoscopy and other observations during transport may have contributed to the fatal outcome in one case. Careful planning of neonatal transports according to listed recommendations and considering the specific problems in each case may prevent complications. PMID- 8421853 TI - [Social and economic course after amputation of the upper extremity]. PMID- 8421854 TI - [Double-blind dose-response study of zidovudine in AIDS and advanced HIV infection. Nordic Medical Research Councils' HIV Therapy Group]. PMID- 8421855 TI - [Botulism in Ammassalik]. AB - Botulism is a rare and serious form of food poisoning and was diagnosed for the first time in the East Coast of Greenland. Historical reports suggest that outbreaks of this condition have occurred previously in this region. In 1990, however, the presence of Clostridium botulinum type E could be confirmed with certainty. Eight individuals partook of a meal which consisted of raw seal meat and raw seal intestines. Four of these developed symptoms of botulism and two of these required assisted ventilation. On the basis of this experience, the medical officers of health in Greenland recommend that all hospitals in Greenland should maintain a supply of antitoxin. PMID- 8421856 TI - [Oxygen therapy at home. A review of the frequency of oxygen use for therapy at home in Denmark]. PMID- 8421857 TI - [Brain biopsy in AIDS patients--is it justified in clinical practice?]. PMID- 8421858 TI - [Registration of traumatic hip dislocation in a nationwide registry]. PMID- 8421859 TI - [Silicone breast prosthesis and connective tissue diseases]. PMID- 8421860 TI - [Siberian psychiatry--prison camps?]. PMID- 8421861 TI - [Antihypertensive treatment. Benefits and costs]. PMID- 8421862 TI - [Life-threatening group A streptococcal infections]. PMID- 8421863 TI - [Toxic shock syndrome in group A streptococcal infection]. AB - Three new cases of toxic shock syndrome due to infection with group A beta hemolytic streptococci are described and similar cases in the literature are reviewed. The typical features of this disease include rapid development of multiorgan failure with renal impairment and, in many patients, also the respiratory distress syndrome. Cardiac dysfunction with myocardial depression is a prominent feature which is most reasonably explained by an effect of the septicaemia per se but may also be toxic cardiomyopathy mediated by circulating toxins. Other major findings include exanthema--often with the development of haemorrhagic bullae as part of toxic epidermal necrolysis. In patients with initial soft tissue infection this is rapidly progressive and often associated with necrotizing fasciitis and myositis, which may give rise to a compartment syndrome with rhabdomyolysis. In addition to conventional therapy with antibiotics, fluid replacement and inotropics, most patients with extensive soft tissue infection also require surgical intervention with debridement and occasionally fasciotomy. PMID- 8421864 TI - [Silicone breast implants and connective tissue disease]. AB - The American Food and Drug Administration (FDA) recently introduced a temporary stop for silicone breast prosthesis implants. The reason is a suspected connection between silicone implants and the development of autoimmune disease. The present authors have reviewed the 32 cases of connective tissue disease which have hitherto been reported in the literature, of these 13 were cases of systemic sclerodermia. On the basis of approximately 2 million silicone prosthesis implants which were introduced in USA alone during the past 20 years, approximately 200 cases of systemic sclerodermia would be anticipated. Nevertheless, the fact that several patients had spontaneous remission of their chronic disease after removal of the silicone implant speaks in favour of a possible connection. It is concluded that on account of the limited number of cases of connective tissue disease which have been reported in patients with silicone implants, insufficient evidence is present to stop implantation of these. Implantation of prostheses filled with saline are recommended for patients with connective tissue disease. PMID- 8421865 TI - [Endoscopic prosthetic treatment in stenosis of the choledochus caused by malignant disease]. AB - Endoscopic insertion of a biliary endoprosthesis is often the treatment of choice in patients with malignant common bile duct obstruction. In experienced centers the procedure is combined with low risk and high success rates. A well known and unresolved problem is the tendency of the stents to clog within a few months, recessitating their replacement. The best survival of the stents is achieved by using the largest straight teflon stent without sideholes. Long-term treatment with antibiotics, antimicrobial impregnated stents, stents without side holes or intraductally placed stents might increase the survival of the stents. Great experience is necessary in order to ensure optimal endoscopic treatment of all the patients with malignant common bile duct obstruction and the treatment therefore should be performed in few large centers. PMID- 8421866 TI - [Sick-listing in pregnancy. Extent, diagnostic pattern and the issuing authority]. AB - Out-of a study population of 1127 women who were delivered at Sct. Maria Hospital, Vejle 993 (88.1%) had had relations with the labour market during their pregnancy. 43% of these 993 women were sicklisted in average 8.4 weeks apart from their legal or agreed leave of four or eight weeks prior to the expected date of delivery. The diagnostic panorama was dominated by four diagnoses: 1) Bleeding, 2) threatening premature birth, 3) back pain or 4) symphysiolysis. These diagnoses could account for half the total number of weeks of sicklisting, and were involved in one quarter of the women sicklisted. In accordance with the intended role of the family physician, the majority of the sicklistings were issued by this instance. PMID- 8421867 TI - [Utilization of the right to maternity leave and resumption of work after delivery. A longitudinal prospective study]. AB - A longitudinal interview study of 360 mothers was carried out to investigate: 1) utilization of maternity leave by the parents, 2) mothers' resumption of occupation after delivery and 3) duration of breast feeding seen in correlation with the mothers' resumption of work. 1) 95% of the families utilized the entire maternity leave. In 92% of the families, the mothers utilized all 24 weeks and, in 3% of the families, the fathers took part in the maternity leave. Only 46% of the fathers utilized the right to take leave in connection with delivery and 14% employed their holiday but were at home for a significantly shorter period. 2) Only one fourth of the mothers had resumed work 24 weeks after the delivery. 44% of the mothers prolonged the maternity leave with their holiday and 32% prolonged the maternity leave even longer. One year after delivery, significantly more mothers were housewives than previously. 3) A significantly positive correlation was found between the time for maternal return to work and the duration of breast feeding. We conclude that almost all of the families utilized the maternity leave and that prolongation of maternity leave (which is 24 weeks after delivery at present) might be necessary. PMID- 8421868 TI - [Course of pregnancies and labors in a department of general surgery. A prospective study of pregnancies during a 1-year period at Horsholms hospital]. AB - The complications of pregnancy and delivery in a material consisting of 841 women without obstetric risk factors were investigated. The deliveries were planned to take place in a department of general surgery with a specialist obstetrician. Seventy-five women (8.9%) were transferred to a specialized obstetric department before delivery, 14 women (1.7%) because of imminent premature delivery. In addition, 69 women were transferred because of removal or their own wish for transfer. A total of 631 patients (90.5%) were delivered vaginally and 66 (9.5%) were delivered abdominally. In thirty-five of these, caesarean sections were performed during the course of delivery. In 34 deliveries (5.1%) vacuo extraction was used. Among 181 women (27.1%), unexpected complications occurred, 80 of these women (12.0%) requiring emergency treatment. A total of 697 infants were delivered. One infant died before delivery because of placental separation. Twenty-two infants (3.2%) were transferred to a paediatric department during the neonatal period, three because of asphyxia. Three of the liveborn infants (0.4%) had Apgar scores at 1 minute between 0 and 3, 40 infants (5.7%) had scores between 4 and 6. After 5 minutes, no infants had Apgar scores between 0 and 3, but three infants had scores between 4 and 6. It is concluded, that the low perinatal mortality and morbidity might indicate, that deliveries in a department of general surgery are safe, provided a specialist obstetrician is available and the rules of referral strictly observed. PMID- 8421869 TI - [Danish experience with physical and occupational rehabilitation after heart transplantation. The heart transplantation group at Rigshospitalet]. AB - Heart transplantations have been carried out for one year (1.9.1990-1.9.1991) in Denmark. Twenty-three out of 27 patients survived at the end of this period. Prior to transplantation, all of the patients were in NYHA groups III or IV. On discharge, all of the patients could manage a 45-60 minutes training programme followed by a stair test (two to six floors up). Prior to transplantation, 17 patients received financial aid in one form or another and one child received special schooling. On an average 165 days (1.9.1991) after transplantation, nine patients were in full or part-time employment, two were students, seven received financial aid and five were still in hospital. A correlation between the duration of financial aid before transplantation and return to work after transplantation was found. In addition, age was found to be of significance. It is concluded that physical status and return to work are satisfactory. PMID- 8421870 TI - [Low back pain and back exercise]. AB - In the last decennium, information regarding the efficacy of back exercises in the treatment of back pain has increased. It has been shown that exercise programs in acute back pain prevents chronicity, and that back exercises in chronic patients decreases back pain and disability. It is primarily the high intensity and duration of the exercise program which is the most important for a successful result rather than the design of the exercises. PMID- 8421871 TI - [Risk factors for development of low back pain]. AB - The incidence, severity and potential disability of low back pain are related, in particular, to previous back complaints, heavy lifting and the general psychosocial environment and to the physical constitution of the individual, viz the endurance of his back muscles. Recommendations for approach to the problem in future are discussed. PMID- 8421872 TI - [Hemodynamic aspects of thoracic epidural analgesia]. AB - Twenty-seven experimental and clinical studies on hemodynamic changes during thoracic epidural analgesia (TEA) are reviewed. It is concluded, that TEA exerts negative chronotropic, inotropic, dromotropic and bathmotropic effects on the heart. The systemic vascular resistance as well as cardiac output are diminished, and a substantial reduction in the myocardial oxygen consumption is also seen. Under close monitoring of the cardiovascular status, TEA seems suitable for surgery in patients with ischemic heart disease. Furthermore, the use of TEA with moderate dosage of the local analgesic agent could be a supplement in the treatment of severe angina pectoris. PMID- 8421873 TI - [Is it possible to establish a constant physician/nurse team at an emergency department of internal medicine?]. AB - In order to obtain better continuity in the contact between the patient and doctors/nurses during hospitalization in a medical department, small teams of nurses and doctors were established, so that the patients were attached to their personal doctor and nurse. In nearly three-fourth of the weekdays the patients saw their personal doctor and nurse. We conclude, that within the Danish public health care system and according to the contractual agreements it is possible to establish small doctor/nurse teams and by this to obtain a high continuity during daytime. PMID- 8421874 TI - [Does the patient obtain close contact with the hospital physician when a physician/nurse team is established?]. AB - The contact between patients and doctors was examined having established smaller doctor/nurse teams to take care of the same seven to eight patients during the weekdays of their stay at hospital. Ten weekdays after their admission to hospital 63% of the patients had not had contact with any other doctor(s) than their personal doctor(s), and 70% of the patients had seen only two different doctors. During the same period the doctors saw their patients three times on an average. We conclude that organizing the hospital doctors in small teams is a way to obtain good continuity in the contact between the patient and the hospital doctor. PMID- 8421875 TI - [Attitude of students to physician's health behavior--an analysis of attitudes and behavior]. AB - A total of 419 students in the first half of their curriculum (243 medical students, 39 engineering students, 36 sociology students and 101 law students) replied to a self-administered questionnaire during autumn 1988 and spring 1989. The questionnaire was concerned with health and included, among other things, their attitudes to doctors' health behaviour. The objects of the questionnaire were 1) to reveal the number of restrictive students who consider that doctors should show a good example as regards health behaviour e.g. as regards smoking, alcohol consumption, diet and physical exercise), 2) to characterize the social profiles of the restrictive students, 3) to assess whether the attitudes and behaviour are in agreement and 4) to assess whether the proportion of restrictive medical students differs from the other students. The result of the questionnaire reveals that 44.4% of all the students were restrictive. The social profiles of these students showed a slight difference from those of the non-restrictive students but only as regards variables related to health behaviour. No difference was found in the degree of restrictiveness in medical students and the remaining students. In addition, the investigation revealed that there was only a slight association between attitudes to doctors' health behaviour and their own health behaviour. PMID- 8421876 TI - [Transesophageal echocardiography and endocarditis]. AB - In 49 patients (34 men and 15 women with an average age of 51 years, range 21-81 years) with a total of 51 episodes of suspected or already demonstrated endocarditis, the diagnostic and therapeutic value of transthoracic echocardiography (TTE) was compared with transoesophageal echocardiography (TEE). It was demonstrated by operation, autopsy, or the course of the condition, that endocarditis was present in 34 cases, while 17 patients did not have endocarditis. The correct diagnosis was established in 19 out 51 cases (37%) by TTE and in 44 (85%) cases by TEE (p < 0.05). The number of ambiguous investigation results fell significantly from 30 (58%) with TTE to seven with TEE. A total of 14 cases of cavity formation related to endocarditis, rupture of fistulae, or perivalvular leakage from prostheses occurred. Three (21%) of these complications were demonstrated by TTE while TEE demonstrated all 14. After examination with TEE, treatment of the patients was changed in 20 cases (39%). It is concluded that: 1) TEE can confirm or exclude the diagnosis of endocarditis with much greater certainty than TTE, 2) TEE more than halves the number of ambiguous results of investigation and 3) TEE multiplies recognition of complications of endocarditis. Even although the results from the cardiological/thoracic surgical centre cannot be transferred to the primary hospital just like that, the results of these and other investigations suggest that TEE should be carried out when TTE cannot confirm or exclude clinically suspected endocarditis with certainty.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421877 TI - [Pattern of injuries at a Danish sports college]. AB - A batch of college students was followed with the object of describing the pattern of athletic injuries. Men sustained the greatest number of acute injuries while no sex difference was observed in the number of stress injuries. The majority of injuries occurred in connection with gymnastics but, when all ball games are considered together, ball game injuries exceeded the gymnastic injuries in number. This difference is further increased when the fact that, in this college, twice as much time is spent on gymnastics as on ball games, is taken into consideration. PMID- 8421878 TI - [Efficiency of eye shields during phototherapy]. AB - The penetrance of light through 21 different types of eye shields used in Danish neonatal care units was evaluated. A phototherapy unit (Drager Phototherapiegerat 800) with a visual illuminance of 8317 Lux was placed in a distance of 39 cm from a photographic lightmeter (Minolta Autometer IIIF). The opening of the meter was covered by the various shields tested. The penetrance varied between 1.0 and 2886 Lux. The most effective shield was made of three layers of green, tightly woven cotton. A commercially made eye shield (Posey Eye Protector 4646) was able to reduce visual illuminance to 118 Lux (ambient light in our nursery was 20-139 Lux). A tubular stockinette covering was found to be ineffective as thicknesses of 1-16 layers was able to reduce penetrance to only 2886-1055 Lux. The use of an eye protector made of three layers of green cotton, sized 140 x 50 mm is recommended. PMID- 8421879 TI - [Ejaculation induced by electrostimulation in men with spinal cord injuries]. AB - Five men (aged 29-43 years) with ejaculatory dysfunction and spinal cord injuries for 10-41 years and level of complete lesions from C6 to T10 underwent electrostimulation. Antegrade ejaculate was obtained in all five participants and simultaneous retrograde ejaculation in four. In general, the total number of spermatozoa was greatest in the retrograde ejaculates and highly variable between the subjects both in the antegrade (0.2-5330 mill.) and in the retrograde ejaculates (0.4-5690 mill.). The percentage of immobile and morphologically abnormal spermatozoa was increased compared with normal values. In connection with the electrostimulation procedures no cases of autonomous hyperreflexia or rectal mucosal injury were observed. PMID- 8421880 TI - [Surveys of diet costs in patients with diabetes mellitus]. PMID- 8421881 TI - [Treatment of asthmatic bronchitis in small children with steroid inhalation]. PMID- 8421882 TI - [Venoruton in chronic venous insufficiency]. PMID- 8421883 TI - The role of uropathogens in feline lower urinary tract disease. Clinical implications. AB - Bacterial, fungal, and parasitic uropathogens have small but significant roles as causative agents in naturally occurring feline lower urinary tract disease. However, the exact cause of hematuria, dysuria, and/or urethral obstruction remains unknown in a large percentage of cats. Feline calicivirus, feline syncytia-forming virus, bovine herpesvirus-4, mycoplasmas, and ureaplasmas are potential uropathogens whose etiopathogenic roles in idiopathic feline lower urinary tract disease remain, as of yet, unresolved. PMID- 8421884 TI - Feline dirofilariasis. AB - The cat must be recognized as a different entity than a dog when considering heartworm disease. The cat responds differently than the dog to larval migration, adult development, and therapy. As a result of these differences, the clinical signs shown by the cat and the therapeutic alternatives must be considered as unique to the cat. The diagnosis of heartworm disease in cats usually is more difficult than in the dog; therefore, the veterinarian must be more aware of the strengths and weaknesses of thoracic radiography, microfilaremia tests, and serologic tests than in the dog. Clinical signs in the cat are usually more vague and generalized than in the dog. When a diagnosis of heartworm disease in cats is made, the decision of which therapeutic regimen to use is a challenge. In many instances, the best treatment (except for supportive treatment) may be no treatment. Knowledge of the incidence of heartworm disease in an area may determine whether a veterinarian counsels a client to consider one of the preventive medications. PMID- 8421885 TI - Feline gingivitis-stomatitis-pharyngitis. AB - Inflammatory conditions of the feline mouth are commonly encountered in small animal practice. Although the majority can be attributed to dental disease and a small percentage are due to autoimmune diseases, the eosinophilic granuloma complex, neoplasia, and other miscellaneous syndromes, many cases appear to be due to a gingivitis-stomatitis-pharyngitis complex, which is likely multifactorial in origin. Viruses, bacterial infection, diet, dental disease, oral conformation, genetic predisposition, hypersensitivities, immunoinsufficiencies, and other defects in oral defense mechanisms may all be contributory. The complexities of this syndrome have made it one of the most challenging diagnostic and therapeutic problems in feline medicine. PMID- 8421886 TI - Nutritional management of cats with infectious disease. AB - The effect of malnutrition in patients with a variety of ailments is known to be deleterious to their overall quality of life, response and tolerance to therapy, and ultimate survival. These effects may be amplified in feline patients with infectious disease owing to the blunting of immunologic responsiveness that results from malnutrition. This vicious cycle of infection, ensuing malnutrition, and subsequent immunocompromise may not allow the patient to mount an attack on the invading pathogen adequately. The most logical means to reverse this cycle is to ensure an adequate nutritional state in the patient. Initial attempts at maintaining or increasing voluntary oral intake should be initiated in the short term. However, enteral or parenteral support should be instituted as quickly as possible if these should fail. Much is yet to be discerned about specific optimal supplementation techniques and dietary formulations for feline patients; however, the application of relatively simple and general principles of nutritional support offer our feline patients with infectious disease the best opportunity to overcome their disease. PMID- 8421887 TI - Role of infectious agents in respiratory disease. AB - There is no question that disease prevention, rather than treatment, represents the single most significant advancement in the management of infectious diseases in companion animals. In particular, the introduction of sensitive, rapid in clinic diagnostic test kits, new vaccines, and implementation of strict vaccination protocols has dramatically decreased the morbidity and mortality associated with viral infections in cats. However, neither the agents nor the disease they cause will be eradicated through the use of even the most comprehensive testing and vaccination programs alone. In 1908, W. T. Sedgwick wrote in his leading public health textbook that "the absence of dirt is not merely an esthetic adornment." Comments such as this, published by noted public health authorities at the turn of the century, marked the beginning of the health revolution in the United States. As the nineteenth century ended, mortality rates from infectious diseases among infants and young adults dramatically and abruptly plunged. The fact that personal hygiene was the hero of this revolution illustrates the importance of the fundamental rules of cleanliness in disease prevention that still apply today. Although basic to infection control, ignoring even the simplest concepts of cleanliness and hygiene in the environment of the multiple cat household can lead to diastrous consequences. PMID- 8421888 TI - Intestinal protozoa infections. AB - This article reviews important intestinal protozoal infections in cats. Among intestinal protozoa, Giardia and coccidia are the most important infections. Giardiasis is a common problem in catteries, especially where young kittens run free on the floor. Overcrowding and high humidity favour the survival and transmission of Giardia. Diagnosis is made by fecal examination by direct or concentration methods. Metronidazole is the drug of choice for treating giardiasis. Among the coccidial parasites, Isospora felis and Cryptosporidium sp. are the common infections. Virtually all cats become infected with Isospora felis. The pathogenicity of I. felis is controversial. Cryptosporidium parvum infection is a zoonosis. Cryptosporidium is transmitted by direct fecal-oral cycle. Cryptosporidial infections appear to be clinical in immunosuppressed cats. No drug has been found satisfactory for treatment of cryptosporidiosis. PMID- 8421889 TI - Feline zoonotic diseases. AB - Many of the feline zoonoses occur more frequently in veterinary personnel owing to their direct contact with cats and the potential for exposure to infected body tissue or fluids. Infection of humans with Afipia felis, Yersinia pestis, Francisella tularensis, and other aerobic/anaerobic bacteria may cause great discomfort and in some situations terminal illness. Although many systemic fungal agents infect humans and cats, only Sporothrix schenckii has been shown to infect humans following direct exposure to infected cats. Various parasites, enteric protozoans and bacteria, and Toxoplasma gondii infections also may cause significant human illness. Therefore, routine handling of cats may expose human personnel in a veterinary facility to an array of important or emerging feline associated human illnesses that occur in the United States. PMID- 8421890 TI - Feline hemobartonellosis. AB - H. felis is a rickettsial parasite that causes hemolysis and sequestration of feline erythrocytes. It should be considered as a potential primary pathogen or opportunist in any cat presented with signs ranging from episodic malaise to acute anemic collapse. Diagnosis requires visualization of the organism in properly prepared blood smears. Treatment uses antirickettsial drugs, corticosteroids, and supportive measures. Clinical recovery requires immune containment of the organism. Treatment does not eliminate the organism from the host. Carrier cats may relapse when their immunity is severely compromised by other diseases such as FeLV. Transmission is presumed to be by blood-sucking parasites and possibly bite wounds between cats. Prevention requires prudent health management of cats. Future advances in the knowledge of the disease will relate mainly to the development of a diagnostic technique that will allow identification of all infected cats. PMID- 8421891 TI - Feline neonatal sepsis. AB - When bacterial infections exceed or overcome the ability of a kitten's immune system to provide protection, life-threatening illnesses such as neonatal sepsis often occur. Many kittens with neonatal sepsis show unusual presentations or a wide variety of clinical presentations that may not be immediately recognized as being associated with sepsis. Because neonatal sepsis causes unexpected sudden death, kittens suspected of having sepsis should be treated immediately. In most instances, initial antimicrobial therapy is selected empirically. Kittens are treated by giving intravenous or intraosseous fluids for dehydration, oxygen to counter tissue hypoxemia, and glucose if hypoglycemia is present. The beta-lactam antimicrobial agents such as the penicillins, cephalosporins, and the combination of beta-lactam antimicrobials and beta-lactamase inhibitors are considered to be the first choice in the treatment of any septicemic kittens. PMID- 8421892 TI - A mutation of cauliflower mosaic virus gene I interferes with virus movement but not virus replication. AB - A 300-bp deletion was made in gene I of a multimeric copy of cauliflower mosaic virus (CaMV). The deleted and wild-type constructs were mobilized into Agrobacterium tumefaciens and used for the inoculation of plants and leaf discs of the host plant, turnip, by "agroinfection." Infection induced by the wild-type construct gave characteristic viral symptoms on the inoculated and systemically infected leaves of whole plants whereas no infection was detected using the deletion mutant. The possibility that the deletion in gene I affected a cis related function (e.g., encapsidation) was tested by complementation of mutant gene I with wild-type CaMV. Double infection experiments showed that replication and spread of both viruses occurred without detectable recombination of the mutant. In contrast to the situation in plants, agroinfection (Grimsley et al., 1986, Proc. Natl. Acad. Sci. USA 83, 3282-3286) of leaf discs, detected as the accumulation of encapsidated forms of virion DNA, was recorded for both the wild type and the deleted constructs. The reduced level of virus accumulation from the mutant was consistent with virus replication occurring only in cells involved in the primary interaction with Agrobacterium, and accordingly progeny viral DNA was only detected in cells around the edge of the tissue disc. These data provide definitive evidence that CaMV gene I encodes a movement protein and show that this function can act in trans in a virus infection. PMID- 8421893 TI - Stable expression of the transfected HIV-1 env gene in a human B cell line: characterization of gp120-expressing clones and immunobiological studies. AB - The env gene of human immunodeficiency virus type 1 (HIV-1) encodes gp 120/41 which plays an important role in the viral infection process and pathogenesis. The surface glycoprotein gp120 is a candidate molecule for the development of a subunit vaccine against HIV-1-induced acquired immunodeficiency syndrome (AIDS). However, thorough studies on the immunobiology of this molecule are hampered by the lack of a suitable model. With this background in mind, and in order to learn more on anti-gp120 cellular immunity, we attempted to develop gp120-expressing human cell clones. Thus by transfecting a human lymphoid cell line of B lineage (Raji), which is known to be resistant to the natural killer cell activity, with an expression vector encoding the envelope and vpu, we established three clones that stably express gp120/41 and vpu. The surface glycoprotein gp 120 is also expressed on the cell surface of these clones. The transfected cells from syncytia with CD4+ human cell lines as well as with peripheral blood mononuclear cells (PBMC) leading to the death of the fused cells. This observation represents additional evidence for the eventual depletion of CD4+ viral targets that fuse with adjacent HIV-infected, gp 120-expressing cells. The latent Epstein-Barr virus genome present in the transfected cells, was not induced to express the lytic cycle antigens. The densities of the surface expression of a number of molecules examined remained unchanged in the transfected cells except for the surface IgM, which increased significantly (P < 0.05) in two clones. One of the clones exhibited a significantly (P < 0.05) reduced proliferation rate as compared to the other clones. The transfected cells of all the three clones showed a significantly (P < 0.01) increased susceptibility to lysis by the PBMC from normal, healthy individuals in a 16-hr 51Cr-release assay. This is the first report of the MHC- and antibody-independent lysis of human cells transfected with the HIV-1 surface glycoprotein. The transfected cells also served as targets in a gp120-specific antibody-dependent cellular cytotoxicity assay. We anticipate that the present model will prove very useful for studying the gp120-specific immune responses in HIV-infected individuals. PMID- 8421894 TI - The matrix region is responsible for the differential ability of two retroviruses to function as helpers for vector propagation. AB - We have investigated the ability of two related reticuloendotheliosis viruses to propagate a spleen necrosis virus (SNV) based retroviral vector in canine osticosarcoma (D17) cells. Reticuloendotheliosis virus strain A (REV-A) consistently propagated the vector more efficiently than SNV in cell culture. To identify the area of the viral genome responsible for the superior helper function of REV-A, we constructed chimeric viruses between SNV and REV-A. Analysis of helper function indicated that a virus comprised of the SNV genome, but containing the matrix region of REV-A, could propagate the vector as well as REV-A. Although REV-A is also a superior virus for vector propagation in chicken embryo fibroblast cells, the region of the viral genome that confers superior helper function does not map to the gag region of REV-A in this cell type. PMID- 8421895 TI - The beet western yellows virus ST9-associated RNA shares structural and nucleotide sequence homology with carmo-like viruses. AB - Complementary DNAs corresponding to the virion RNAs of the ST9 isolate of beet western yellows virus (ST9 BWYV) were cloned and the complete, 2843 nucleotide sequence of a unique RNA associated with ST9 BWYV, the ST9-associated RNA (ST9 aRNA), was determined. Three ORFs with deduced translation products of molecular weights 22,600, 12,500, and 35,900, respectively, and a small ORF, potentially encoding for a 3400 molecular weight protein, were identified in the ST9 aRNA sequence. Possible gene products of the ST9 aRNA were identified by in vitro translation analysis of the ST9 BWYV virion RNAs (the ST9 BWYV genomic RNA [ST9 BWYV gRNA] and the ST9 aRNA) and of transcripts representing the nearly full length ST9 aRNA, individual ORFs, and specific ORF combinations. Proteins of 23,000 and 28,000 M(r) were derived from ORF 1 transcripts, while proteins of 25,000 and 35,000 M(r) were derived from ORF 3 transcripts. Larger proteins of 85,000 and 45,000 M(r) were also identified, and shown to possibly arise by continuous translation of ORFs 1, 2, and 3 (by possible readthrough and frameshift), or ORFs 2 and 3 (by possible frameshift), respectively. No products were detected from transcripts representing only ORF 2. Computer database searches and analyses indicated that the ST9 aRNA shares significant nucleotide sequence homology with carmo-like viruses, but not with its helper virus, BWYV. Amino acid sequence analyses also indicated that the ST9 aRNA ORF 2 and ORF 3 regions share extensive homology with putative RNA-dependent RNA polymerases of carmo-like viruses. PMID- 8421896 TI - High-level expression of the Japanese encephalitis virus E protein by recombinant vaccinia virus and enhancement of its extracellular release by the NS3 gene product. AB - Recombinant vaccinia viruses expressing the prM and E genes of the Japanese encephalitis virus (JEV) were constructed by use of synthetic promoters. While the recombinant virus mOJ6-SL, with an optimized vaccinia late-gene promoter, produced a 20-fold elevated level of E protein, as well as an 86-kDa precursor protein in infected cells, no significant quantitative difference was detected between the extracellular or cell-surface E protein produced by mOJ6-SL and those produced by mOJ6 with the 7.5-kDa promoter. However, when the cells were infected with Dengue 2 virus before infection with mOJ6-SL, the amount of the extracellular E protein increased 16-fold. In addition, enhancement of its extracellular release was observed when cells were co-infected with mOJ6-SL and recombinant vaccinia virus expressing the NS3 gene of JEV. PMID- 8421897 TI - A general method for the identification of transcribed retrovirus sequences (R-U5 PCR) reveals the expression of the human endogenous retrovirus loci HERV-H and HERV-K in teratocarcinoma cells. AB - During the past decade, different types of endogenous retroviral sequences have been defined in the human genome usually by low stringency hybridization employing DNA probes of evolutionary conserved animal retrovirus genes. Although all human genomic loci sequenced were found to be defective or interspersed with stop codons, indirect evidence is accumulating that human endogenous retroviral loci are expressed at least in some instances. One example is the synthesis of retroviral particles in human teratocarcinoma cell lines observed by electron microscopy. To establish a link between virus expression and genomic loci we searched for retroviral RNA in human cellular mRNA populations using a generally applicable method. A tRNA-derived primer complementary to a putative retroviral primer binding site was extended by reverse transcription and this product was elongated with a homopolymeric stretch and amplified by PCR (R-U5 PCR). Cloning and sequencing of such products revealed that the endogenous retroviral loci HERV H and HERV-K are expressed in those human teratocarcinoma cell lines which produce retroviral particles. The size distribution of four HERV-K mRNAs detected in Northern blots is reminiscent of the complex expression pattern seen with a number of exogenous retroviruses. PMID- 8421898 TI - Reproducible high level infection of cultured adult human hepatocytes by hepatitis B virus: effect of polyethylene glycol on adsorption and penetration. AB - We have previously succeeded in infecting normal human hepatocyte primary cultures with hepatitis B virus (HBV). However, infection was subject to individual variations even in the presence of dimethyl sulfoxide (DMSO), which appeared to increase the amounts of viral DNA associated with the cells. In this study, we have defined conditions which enhance hepatitis B virus penetration into the cells, and we show that, under these conditions, infection of hepatocytes is always possible, regardless of their individual origin. We have found that addition of polyethylene glycol (PEG) to the cultures maintained in the presence of 2% DMSO at the time of infection markedly increased the infection process and made it highly reproducible. Moreover, both the tissue and species specificity were preserved. This increased HBV infection was correlated to increased amount of internalized HBV DNA and to enhanced attachment of the virions. From these results it may be assumed that PEG could favor a better interaction between virions and cells, resulting in an activated internalization of bound viral particles. Data also show that adult human hepatocyte primary cultures, which are not equally susceptible to HBV infection, are consistently capable of viral replication when the viral genome has entered the cells. This suggests that the main limitation of the in vitro HBV infection lies in the ability of human hepatocytes to specifically bind the viral particles. PMID- 8421899 TI - Repair in vivo of altered 3' terminus of cymbidium ringspot tombusvirus RNA. AB - Progeny RNA of CyRSV RNA transcripts ending with -GGG, instead of -CCC as in wild type RNA, was analyzed and shown to revert to wild-type. A new full-length cDNA clone of CyRSV was prepared, bearing the correct 3' terminus, from which transcripts could be synthesized having very high infectivity. Mutants were prepared which lacked the three terminal Cs; transcripts were infectious and most cDNA clones recovered from progeny RNA had regained the wild-type sequence. Transcripts ending in -GGCCAn were also infectious, and the 3' sequence of most cDNA clones recovered after inoculation was the same as the inoculum sequence. Deletion of G at position -4 completely abolished infectivity. The occurrence of a repair mechanism at the 3' end of CyRSV similar to telomerase activity on chromosomes is suggested. PMID- 8421900 TI - Enhanced production of morbillivirus gene-specific RNAs following induction of the cellular stress response in stable persistent infection. AB - Previous in vitro work demonstrated the incorporation of the major inducible 70k heat shock protein (i.e., 72k HSP) into the biologically active light nucleocapsid (L-NC) variant of canine distemper virus (CDV). Here, in vitro induction of the cellular stress response, characterized by elevated cytoplasmic and intranuclear 72k HSP, enhanced L-NC expression in mink lung cells supporting stable persistent infection by raccoon-origin CDV. Increases in L-NC were correlated to increased viral RNA production in cell-free transcriptional assays. The enhanced production of viral transcripts within infected cells following stress response induction was confirmed by slot blot and Northern blot analysis of total cellular RNA and was reflected in increased total viral protein production. Post-shock increases in viral fusion (F) gene transcripts and F protein were associated with dramatic increases in viral cytopathic effect. Modest induction of cell-free infectious viral progeny was also documented. A similar effect of the cellular stress response upon viral protein expression, cytopathic effect, and cell-free infectious progeny release was demonstrated in murine neuroblastoma cells persistently infected with a canine CDV isolate. Alterations of the persistent viral phenotype were independent of the specific mechanism of stress-response induction (i.e., heat or sodium arsenite), supporting the role of the stress response and not a particular stressor in mediating these changes. These results document the ability of the cellular environment to alter persistent viral RNA metabolism, thereby altering the infection phenotype. PMID- 8421901 TI - Mutagenesis of conserved residues at the yellow fever virus 3/4A and 4B/5 dibasic cleavage sites: effects on cleavage efficiency and polyprotein processing. AB - Flavivirus proteins are produced by co- and post-translational proteolytic processing of a large polyprotein using both host- and virus-encoded enzymes. The flavivirus serine proteinase, which consists of NS2B and NS3, is responsible for cleavages of at least four dibasic sites in the nonstructural region. In this study, a number of substitutions for the conserved amino acids flanking the 3/4A and 4B/5 dibasic cleavage sites [Arg(P2)-Arg(P1) decreases Gly(P1')] were examined for their effects on yellow fever virus (YF) polyprotein processing. The substrate for these studies was a truncated YF polyprotein, called sig2A-5(356), which consists of a signal sequence fused to NS2A and extending through the first 356 amino acids of NS5. At the P1' position (Gly) of the 4B/5 site, only Ser and Ala were allowed while six other substitutions abolished cleavage. Substitutions of the 4B/5 P1 Arg residue with Lys, Gln, Asn, or His were tolerated while replacement with Glu eliminated cleavage. The 4B/5 P2 position (Arg) was found to be tolerant of substitutions with polar or hydrophobic residues which allowed varying degrees of partial cleavage. Previous studies have shown that cleavage at the 3/4A site is incomplete in YF-infected cells and that the cleavage efficiency at this site is significantly less for the sig2A-5(356) polyprotein. Replacement of the 3/4A P1 Arg residue with noncharged polar or hydrophobic residues reduced the cleavage efficiency, whereas substitutions with Glu or Pro abolished cleavage. Studies with polyproteins containing one or both of the 3/4A and 4B/5 cleavage sites blocked indicate that there is not an obligatory processing order for cleavages generating the N termini of YF NS4A, NS4B, and NS5. PMID- 8421902 TI - Analysis of protein expression and virus-like particle formation in mammalian cell lines stably expressing HIV-1 gag and env gene products with or without active HIV proteinase. AB - Cell lines stably releasing noninfectious virus-like particles containing wild type or mutant gene products represent useful tools for a biochemical, immunological, and structural analysis of virus assembly. Human immunodeficiency virus (HIV) type 1 gag and env gene products were transiently and stably expressed in mammalian cells and the formation of virus-like particles incorporating viral glycoproteins was analyzed. Transient cotransfection of plasmids directing the synthesis of gag and env gene products yielded efficient release of particles but specific incorporation of HIV glycoproteins was not detected. A stable cell line expressing wild type HIV-1 glycoproteins was generated and transient transfection of this cell line with gag-encoding constructs led to the release of virus-like particles incorporating HIV surface and transmembrane glycoproteins. Attempts to establish stable cell lines expressing wild type HIV gag and pol genes were unsuccessful and only highly unstable lines primarily expressing uncleaved precursor polyproteins were obtained. This result appears to be caused by the cytotoxic effects of the viral proteinase since stable lines were readily selected after transfection of constructs either encoding an inactive mutant of the proteinase or a mutated frameshift signal which prevented expression of the pol reading frame. Stable coexpression of uncleaved Gag polyprotein and wild type env gene products yielded efficient release of immature virus-like particles incorporating HIV glycoproteins. Electron micrographs revealed lentiviral budding structures with the typical surface projections of viral glycoprotein oligomers. PMID- 8421903 TI - Aurothiolates inhibit HIV-1 infectivity by gold(I) ligand exchange with a component of the virion surface. AB - Aurothioglucose and aurothiomalate have anti-HIV-1 activity in vitro. Antiviral activity requires the formation of a reactive intermediate with a molar equivalent amount of a thiol ligand. This activates gold(I) ligand exchange between the reactive species bis(thiolato)gold(I) and acidic thiol groups exposed on the surface of proteins. Bis(thioglucose)gold(I) (bisAuTG) which is formed by the reaction of molar equivalent amounts of aurothioglucose and 1-thio-beta-D glucose completely protected MT-4 and CEM cells against HIV-1NL4-3-induced cytopathogenicity. Although bisAuTG is an inhibitor of human immunodeficiency virus-1 (HIV-1) reverse transcriptase in a cell-free assay, its antiviral effect is due to modification of a surface component of the virion. The HIV-1 strain NL4 3 is 200-fold more sensitive to inhibition of infectivity by bisAuTG than are the strains MN, RF, and SF-2. HIV-1NL4-3 has a unique cysteine residue close to the amino terminus of its gp41 envelope glycoprotein (residue 532 of gp160) which we hypothesize is the target of bisAuTG binding. Mutation of that residue alters HIV 1NL4-3 infectivity and dominantly suppresses virus assembly when coexpressed with the wild-type NL4-3 genome. We show that bisAuTG treatment releases gp120 from the surface of cells expressing wild-type HIV-1NL4-3 envelope glycoprotein, but it does not release gp120 if Cys532 is mutationally altered to Ala. Thus, the antiviral effect of bisAuTG on HIV-1NL4-3 is due to an effect on the association of gp120 with gp41. PMID- 8421904 TI - Biological activity of cauliflower mosaic virus aphid transmission factor expressed in a heterologous system. AB - Aphid transmission factor (ATF) activity of cauliflower mosaic virus (CaMV) gene II product was recovered after expression of the gene by a baculovirus recombinant. The expression product, when first acquired by aphids through parafilm membrane, was able to mediate the transmission of two aphid nontransmissible isolates (CM1841, CM4-184) providing the first direct evidence that the product of the gene II is the CaMV ATF. The CaMV ATF in its active conformation has a strong tendency to aggregate and all attempts at solubilizing it resulted in the loss of the ATF activity. The CaMV ATF was also expressed in Escherichia coli, using the pGEX 3X plasmid vector, as a fusion protein to glutathione S-transferase (GST) and was purified. The fusion product (GST-P18), whether purified or not, was not able to complement the transmission of transmission-defective isolates. However, when GST-P18 was added to some extracts from a plant infected with an aphid-transmissible isolate (Cabb B-JI), the transmission was inhibited. This suggests that it could be possible to block the in vitro transmission of CaMV using a molecule analogous to the ATF. PMID- 8421905 TI - Gene II product of an aphid-nontransmissible isolate of cauliflower mosaic virus expressed in a baculovirus system possesses aphid transmission factor activity. AB - A new baculovirus expression system was used to express the gene II which encodes the 18-kDa aphid transmission factor (ATF) protein of two isolates of cauliflower mosaic virus (CaMV), the aphid-transmissible (AT+) Cabb B-JI and the aphid nontransmissible (AT-) CM1841 isolates. ATF activity was shown to be associated with the baculovirus expression product of CaMV gene II from both the AT+ and the AT- isolates. No differences could be detected in the stability of the P18s from AT+ and AT- isolates expressed in the baculovirus system. These results indicate that the AT- phenotype of CM1841 isolate might not be due to a defective P18 but to an insufficient concentration of it in infected plant tissue. PMID- 8421906 TI - Analysis of a temperature-sensitive mutation affecting the integration protein of Moloney murine leukemia virus. AB - We have previously described a series of mutations in the 3' terminus of the pol gene of Moloney murine leukemia virus which adversely affect the establishment of an integrated provirus (Roth, M. J., Schwartzberg, P., Tanese, N., and Goff, S. P., J. Virol. 64, 4709-4717, 1990). While most of the mutations were unconditionally lethal for virus replication, we now demonstrate that one mutation, in6161-12, a 12-base pair linker insertion into the 3' end of the pol gene, causes a temperature-sensitive phenotype. This mutant virus is temperature sensitive for IN protein function as well as viral replication. PMID- 8421908 TI - Comparison of structure and sequence of influenza B/Yamagata and B/Beijing neuraminidases shows a conserved "head" but much greater variability in the "stalk" and NB protein. AB - In 1988 the B/Yamagata/16/88 influenza virus appeared, which was antigenically quite different from the other B viruses circulating at that time. Neuraminidase heads of this strain were purified, crystallized, and analyzed by X-ray crystallography and compared with the structure of B/Beijing/1/87 neuraminidase. Only two point mutations could be detected in the "head" of the molecule leading to local rearrangement of amino acid side-chains. The nucleotide sequence, however, revealed significant differences in the stalk region and in the NB protein. PMID- 8421907 TI - The receptor-destroying enzyme of influenza C virus is required for entry into target cells. AB - The hemagglutinin-esterase (HE) protein of influenza C viruses possesses an acetylesterase activity, which appears essential for replication, as determined by reduced infectivity after inhibition of the viral enzyme [Vlasak et al., J. Virol. 63, 2056-2062 (1989)]. Analysis revealed the absence of virus-specific RNA and protein synthesis in infected cells after inhibition of the receptor destroying enzyme. In addition, hemolytic activity was reduced after incubation of influenza C/JJ/50 virus with diisopropyl-fluorophosphate or 3,4-dichloro isocoumarin. Further analysis revealed that inhibition of hemolysis depends on virus and erythrocyte concentrations. It is suggested that an active receptor destroying enzyme is required for entry of influenza C virus into target cells at a step prior to fusion of the viral and cellular membrane. Our data indicate that cleavage of receptors bound to the HE protein is a prerequisite for the low pH triggered conformational change required for fusion. PMID- 8421909 TI - Comparison of the nucleotide and deduced amino acid sequences of the envelope protein genes of the wild-type French viscerotropic strain of yellow fever virus and the live vaccine strain, French neurotropic vaccine, derived from it. AB - The envelope (E) protein genes of the wild-type yellow fever (YF) virus French viscerotropic virus and its' vaccine derivative, French neurotropic virus (FNV), were compared and found to differ by 13 nucleotides that coded for 8 amino acid substitutions. Comparison of the E proteins of FNV and 17D, the vaccine strain derived from wild-type strain Asibi, showed that there was no common nucleotide change or amino acid substitution between these two vaccine strains. However, changes are clustered around amino acid positions 52-56 and may represent the common vaccine epitope shared by 17D and FNV vaccine viruses. The molecular basis of any difference in neurotropism and viscerotropism of YF virus, attributable to the E protein, remains unclear. PMID- 8421910 TI - Modulation of host cell nuclear proteins that bind to HIV-1 trans-activation responsive element RNA by phorbol ester. AB - The trans-activation-responsive (TAR) element located within the 5' untranslated region of HIV-1 mRNA is the cis-responsive element for Tat, the viral trans activator protein. Several TAR RNA binding proteins (TRBPs) have been identified in the nuclear extract from HeLa cells as cellular factors required for a full Tat-mediated trans-activation. In this study, we have tried to identify TRBPs in human T cell line (MOLT-4) persistently infected with HIV-1. Nuclear extract from the infected MOLT-4 cells was analyzed by gel-retardation and uv cross-linking assays with radiolabeled TAR RNA probe. Two major complexes of TAR RNA with some cellular proteins were detected in the gel-retardation assay. As the components of these complexes, at least five TRBPs (p30, p37, p46, p50, and p56) showing specific binding to the TAR RNA were detected in the uv cross-linking assay. We also observed that the detectable levels of p37 and p50 in the infected MOLT-4 cells were greatly reduced after phorbol ester (TPA) treatment under the condition of which HIV-1 gene expression was increased by about fivefold. These results suggest that the modulation of TRBPs by some mitogenic stimuli such as TPA might have a role in the trans-activation of HIV-1 gene expression in vivo. PMID- 8421911 TI - Double cheek-cervical rotation flap for repair of facial defects. AB - Repair of extensive facial defects due to cancer surgery or trauma is a challenge to restore form, function, and facade. This article studies the advantages of using the double cheek-cervical rotation flap for immediate reconstruction of 10 patients with huge facial defects. PMID- 8421912 TI - The use of modern radiologic methods in identifying incidental renal cell carcinoma. AB - The increased use of ultrasonography and computed tomography has made the incidental discovery of renal masses more common. With this factor in mind, the records of 109 patients with renal cell carcinoma diagnosed and treated at Charleston Area Medical Center (CAMC) between 1983 and 1990 were reviewed to determine how many cases were "incidentally diagnosed." Thirty-two (29 percent) of these patients had their renal cell carcinoma discovered incidentally; and of these tumors, 47 percent were Stage I and only 13 percent were Stage IV; for symptomatic patients, 22 percent were Stage I and 36 percent were Stage IV. This study also found that the classic triad of pain, hematuria and flank mass was present in only six of 109 patients (5.5 percent). Tumors found incidentally will usually be of lower pathological stage and may carry a better prognosis. PMID- 8421913 TI - Evaluation of the obstetrical care system in West Virginia. PMID- 8421914 TI - Demands of illness and individual, dyadic, and family adaptation in chronic illness. PMID- 8421915 TI - Transfer of cognitive learning to a clinical skill: linear versus interactive video. PMID- 8421916 TI - Integrating research into an R.N. to B.S.N. clinical course. PMID- 8421917 TI - Ways of discussing validity in qualitative nursing research. PMID- 8421918 TI - Coping with cancer. PMID- 8421919 TI - The phenomena of doing well in people with AIDS. PMID- 8421920 TI - Revision of the Ways of Coping Checklist for a clinical population. PMID- 8421921 TI - The unknown and the unknowable--managing sustained uncertainty. PMID- 8421922 TI - A tribute to "The Mount ". PMID- 8421923 TI - Stress and health outcomes among mothers of low-birth-weight infants. PMID- 8421924 TI - Left-sided leg edema of the elderly: a common variant of the iliac compression syndrome. AB - BACKGROUND: Anatomically, the right common iliac artery crosses the left iliac vein and its accompanying lymphatics. We hypothesized that this situation could lead to a predominance of edema, telangiectasis, and venous varicosities on the left lower extremities of older persons. METHODS: To test this hypothesis, a research assistant who was blinded to the study goals examined 215 predominantly elderly residents of North Carolina homes for the aged and disabled. RESULTS: Among these subjects, 17.7 percent had predominantly left-sided edema, and 5.7 percent had predominantly right-sided edema (P < 0.001). When the 88 subjects with pitting edema greater than 3 mm were studied, 34.5 percent showed a left sided predominance, and 6.9 percent showed a right-sided predominance (P < 0.001). In contrast, no significant difference was found in the lateralization of venous varicosities or of telangiectasis. CONCLUSIONS: Asymmetric edema is common and is usually left-sided in older persons. Compression of the left common iliac vein and its accompanying lymphatics by the right iliac artery, rather than overt clinical disease, might explain the majority of asymmetric edema seen in clinical practice. PMID- 8421925 TI - Attitudes, age, and participation in mammographic screening: a prospective analysis. AB - BACKGROUND: To address the needs of older women, we investigated age-specific attitudes toward mammography that might be influenced by written or verbal communications. METHODS: Attitudinal scores for women aged 40 through 64 years and 65 years and older were calculated prospectively from responses to a mailed questionnaire based on the theory of reasoned action. Age-group mean scores were compared using t-tests for eight components of the attitude measure. Score correlations with participation were compared between age groups using multivariate analysis. RESULTS: Of the 919 eligible women, 666 (72 percent) completed the study questionnaire, and 433 (65 percent) of the 666 women obtained mammograms. A woman aged 65 years or older was less likely to believe that mammography could find a cancer that she (P < 0.01) or her physician (P < 0.05) could not find, and she valued this characteristic less than a younger woman in each instance (P < 0.01). The belief that mammography involved asymptomatic detection was more highly correlated with participation in older women (P < 0.05), as was the attitude that mammography was unfamiliar, but acceptable (P < 0.05). CONCLUSIONS: Older women are less likely to understand that mammography can find cancers that might be missed by other screening methods. Communications to encourage mammography among older women should explain its strengths and familiarize them with the procedure. Communications to younger women need to consider other factors. PMID- 8421926 TI - Families of homosexual men: their knowledge and support regarding sexual orientation and HIV disease. AB - BACKGROUND: The number of homosexual men (gay) with the human immunodeficiency virus (HIV) who will become ill in the next few years is expected to rise dramatically. Many will need and desire support from their families of origin. Understanding the history of family relationships can be crucial for the optimal care of these patients and their families. Little is known, however, about the relationship between gay men and their families of origin. METHODS: A convenience sample of gay men (n = 265) from three northern California cities was surveyed to determine family member knowledge of their sexual orientation and HIV status and perceived family supportiveness regarding issues of HIV disease and acquired immunodeficiency syndrome (AIDS). Fifty-five percent of the sample were HIV negative, 14 percent were HIV positive, 8 percent had received a diagnosis of AIDS, and 23 percent had not been tested. RESULTS: Approximately 70 percent of family members knew a son's or brother's sexual orientation. Fewer family members (50.9 percent) knew a son's or brother's HIV status than knew his sexual orientation. Untested gay men reported the least family knowledge of both sexual orientation and HIV status. There was, however, considerable variation in knowledge among members of the same family, with mothers and sisters knowing HIV status more frequently than fathers and brothers. The amount of supportiveness regarding issues of HIV disease within the family also varied considerably. CONCLUSIONS: It is important for the family physician caring for the gay male patient, his family, or both to understand that the pattern of knowledge and supportiveness among family members concerning sexual orientation and HIV status is selective, even within the same family. The physician needs to assess family members' knowledge and attitudes to plan an overall care strategy. PMID- 8421927 TI - The pesticide-exposed worker: an approach to the office evaluation. AB - BACKGROUND: Pesticide exposures have clinical, epidemiological, legal, and political ramifications that go beyond the confines of a physician's office. The examination of the employee who has been exposed to a pesticide should be undertaken in an organized and methodical manner so that specific questions of causality can be answered and treatment can be initiated. METHODS: Five representative case studies illustrate different circumstances in which a pesticide injury can be seen in the office setting. RESULTS AND CONCLUSIONS: A wide range of pesticides is used in business and home, and the dangers of misdiagnosis, maldiagnosis, underdiagnosis, and overdiagnosis are especially common given the variety of chemicals used. The exposed employee must be removed from the source of exposure. Complete decontamination is a primary concern, and patients with unstable vital signs will need to be hospitalized. A methodical office examination, however, can be carried out on many exposed employees. A detailed description of the circumstances of the exposure should be elicited, and the chemical implicated in the exposure should be researched. A pesticide exposure is a sentinel event in the life of a patient and also suggests that other employees can be exposed. Such an exposure needs to be carefully assessed and documented, and proper treatment must be rendered. Further, the exposure can represent the first of many other exposures that might or might not be reported. Proper notification of authorities can limit exposures before they become severe. PMID- 8421928 TI - Transcervical amnioinfusion. AB - BACKGROUND: Transcervical amnioinfusion is a new and relatively safe, simple procedure that can be performed in most modern hospital maternity units. METHODS: We reviewed the current medical literature concerning this topic by searching MEDLINE files from 1987 to the present, using key words "amnioinfusion," "fetal distress," "premature rupture of membranes," "meconium aspiration," and "oligohydramnios." Older articles were accessed from cross-reference of the more recent publications. RESULTS: When amnioinfusion was used to treat variable fetal heart rate decelerations, it usually reduced the severity of the decelerations, as well as the Cesarean section rate for fetal distress. Prophylactic transcervical amnioinfusion has been studied in three other settings: premature rupture of membranes, meconium passage during labor, and oligohydramnios. A suggested protocol for saline amnioinfusion during labor is given. CONCLUSIONS: Further studies are needed to confirm efficacy reports and to clarify the indications for saline amnioinfusion. PMID- 8421929 TI - The family physician and ethics at the bedside. AB - BACKGROUND: Clinical ethics and ethics committees are a real phenomenon in medicine and play an important role in improving patient care by enhancing the decision-making process between patients and their physicians and thus strengthening this special bond. This article reviews the evolving qualifications of the clinical ethics consultant and compares them with the traditional knowledge, skills, attitudes, and values of the family physician. METHODS: Maternal for this comparison was gathered from a review of the pertinent literature of the past 10 years on the development of clinical ethics and the addition of ethics to the medical curriculum and from personal observations and literature commentary on family practice as a distinct entity. RESULTS AND CONCLUSIONS: Family physicians are uniquely qualified to serve as ethics consultants, particularly with additional training in moral philosophy and health care law. It is hoped that family physicians will become increasingly involved in this growing field to serve as future teachers, researchers, institutional leaders, and policy makers in clinical ethics. More importantly, faculty role models are needed to train family practice residents in ethical decision making at the bedsides of their patients. PMID- 8421930 TI - Detection of major depressive disorder in primary care patients. AB - BACKGROUND: The aim of this study was to assess the utility of a two-phase case detection strategy for major depressive disorder and a quantitative self-rating instrument for dysphoria. METHODS: A convenience sample of 302 ambulatory patients received three self-administered depression rating instruments: a modified version of the Dartmouth Cooperative (COOP) Functional Health Assessment Chart on emotional condition, a three-question screening test for depression taken from the Diagnostic Interview Schedule (DIS), and the Inventory to Diagnose Depression (IDD). All patients' medical charts were reviewed after the administration of these instruments, and those charts of patients found to be depressed were reviewed again 6 months later. A family practice ambulatory patient center in a university hospital was the setting for the study. RESULTS: The IDD detected current major depressive disorder in 41 persons (13.6 percent of sample). Twenty-five of the 41 IDD-positive patients had not previously had a major depressive disorder diagnosed according to chart notes. Six months later, 16 of the 25 patients with newly diagnosed major depressive disorders had not returned to the clinic since the index visit. A single question (DIS question 073b), when compared with the IDD diagnosis of major depressive disorder, had a sensitivity of 95.1 percent, specificity of 78.9 percent, positive predictive value of 41.5 percent, and negative predictive value of 99 percent. Dysphoria, as measured by the modified COOP chart on emotional condition, was significantly worse among those with current major depression (IDD positive) than for those without (IDD negative). CONCLUSIONS: A two-phase case detection strategy for major depressive disorder consisting of a single screening question followed by a self-administered diagnostic instrument can efficiently pick out virtually all ambulatory primary care patients with a major depressive disorder. A brief functional assessment chart can quantify the extent of dysphoria. PMID- 8421931 TI - A comprehensive microcomputer-based medical records system with sophisticated preventive services features for the family physician. AB - BACKGROUND: Computer-based medical records systems improve the provision of preventive services in the offices of family physicians. Until recently, these systems were either not commercially available for use by practicing physicians or were very expensive. METHODS: A commercially available, microcomputer-based medical records system is currently used at the Department of Family Medicine at the Medical University of South Carolina. This system is used as a fully electronic medical record and has sophisticated health maintenance tracking and reminder features. These features track the provision of preventive services, provide physician reminders at the time of patient visits, permit generation of mailed patient reminders, and provide reference to relevant patient education resources. RESULTS AND CONCLUSION: The system described in this paper can be used by practicing physicians to improve their delivery of preventive services. PMID- 8421932 TI - Need for continuing tuberculosis surveillance in previously screened new immigrants. PMID- 8421933 TI - Volvulus during coitus. PMID- 8421934 TI - Patient perceptions of family practice residents in rural private practice settings. PMID- 8421935 TI - Reflections in family practice. The watch. PMID- 8421936 TI - Current report--HIV. Antiretroviral strategies. PMID- 8421937 TI - Clinical ethics and the family physician. PMID- 8421938 TI - Why can't a man be more like a woman? PMID- 8421939 TI - Ambulatory blood pressure monitoring. PMID- 8421940 TI - Pseudoephedrine in pregnancy. PMID- 8421941 TI - Microskills model of clinical teaching. PMID- 8421942 TI - Obstetrics in rural family practice. PMID- 8421943 TI - Autonomic response to beeper. PMID- 8421944 TI - NMR spectroscopy: current status and future possibilities. AB - Nuclear magnetic resonance (NMR) spectroscopy is now established as a non invasive method of studying metabolism in living systems, ranging from cellular suspensions to man. With respect to clinical applications, recent developments include the successful implementation of new techniques for spatial localisation, and in particular the acquisition of excellent 1H spectra from selected regions of the human brain. Localised 1H spectroscopy opens the way to monitoring a wide range of compounds that are inaccessible to 31P NMR, and should add considerably to the information that is available from 31P studies. NMR spectroscopy does, however, have its limitations, which arise primarily from the fact that it is an insensitive technique. This lack of sensitivity limits the spatial resolution for metabolic studies, and means that metabolites must be present at fairly high concentrations in order to produce detectable signals. In this article, we illustrate the scope and limitations of NMR spectroscopy by describing a few examples of studies undertaken on animals and humans. PMID- 8421945 TI - Systematic development of cerebral resuscitation after cardiac arrest. Three promising treatments: cardiopulmonary bypass, hypertensive hemodilution, and mild hypothermia. AB - Since 1970 we have investigated postischemic anoxic encephalopathy and potential treatments for cerebral resuscitation after cardiac arrest by cardiopulmonary cerebral resuscitation (CPCR). The post-resuscitation syndrome has been studied at the levels of cell, organ, organism and community. Short-term and long-term models in rats, dogs, and monkeys have been developed, and an international multicenter randomized clinical trial mechanism was established. Clinical studies disproved the 5-min limit of reversible cardiac arrest and yielded other valuable data on treatments and prognostication. Thiopental loading or calcium entry blocker therapy (lidoflazine) gave no significant improvement in patients. Free radical scavengers are under investigation in the laboratory. We hypothesize that post-arrest perfusion failure and necrotizing cascades require etiology-specific combination treatments. Standard (control) therapy in a current dog model of cardiac arrest (no flow) of 12.5-20 min, reperfusion with cardiopulmonary bypass, and intensive care for 72-96 h has consistently resulted in survival with brain damage. After ventricular-fibrillation (VF) arrest of 17 min, moderate hypothermia (28-32 degrees C) inconsistently improved cerebral outcome. After VF arrest of 12.5 min, hypertension plus hemodilution normalized the local (multifocal) cerebral hypoperfusion post-arrest and, again, inconsistently improved cerebral outcome. Additional mild hypothermia (34-36 degrees C), however, consistently improved cerebral outcome, whether induced before or during and after arrest. PMID- 8421946 TI - Taxonomy of subjective phenomena: a neuropsychological basis of functional assessment of ischemic or traumatic brain lesions. AB - A proper evaluation of functional competence after central lesions has to be based on a classification of functions that one can agree upon. It is a sad fact in neuropsychology that such a classification is not available. An attempt will be made to discuss such a classification (or taxonomy) that might be useful. The basic idea is that elementary psychological functions are evolutionary products whose availability is dependent on the functional integrity of neuronal modules. Such modules are embedded neuronal mechanisms that are linked to localized structures or distributed neuronal algorithms. Constancy of interindividual loss of psychological functions associated with lesions of modules can be used to define a catalogue of functions. Using this principle one can differentiate four areas of psychological functions that are represented in a modular fashion. These areas are stimulus representations ("perception"), processing of information ("learning and memory"), evaluation of information (for instance by emotions), and finally action or reaction. Functional competence is, however, not only described by the potential availability of elementary psychological functions, but also by formal aspects, i.e. how functions are made available. Such formal aspects refer to activation and in particular to temporal problems of neuronal processing. A particular "time machine" will be discussed which is essential for functional competence. Central lesions may either effect the what of functions or the how of functions. A differentiation between these material and formal aspects of functional competence are essential with respect to recovery or restitution of function. PMID- 8421947 TI - Assessment of emergency care in trauma patients. AB - There are many reasons for evaluation of an emergency care system, such as expenses (1.035 Bio. DM in 1985) and quality control. From January 1, 1987 to December 31, 1987 information on all patients seen by an emergency physician in the field have been recorded prospectively in a standard form by the Cologne emergency medical services. Cologne has 1,000,000 inhabitants and covers an area of 405 km2. The patients' status, diagnosis and therapeutic interventions were recorded. Trauma patients were further assessed as to time of accident, cause of accident, and trauma score. All trauma patients with a trauma score < 16 were followed up to their discharge from the hospital. In 1987, 2,073 trauma patients were treated. Overall mortality at the time of discharge was 9.2%. This result alone, however, is not sufficient for assessment of the trauma system. It is important to provide better information on the patient. The trauma evaluation score already used in the US became also a valid instrument in West-Germany. It shows a high correlation between survival and the patients' physiological status in the field. Standard curves could be established for comparing individual or regional trauma systems. PMID- 8421948 TI - Current level of prehospital care in severe head injury--potential for improvement. AB - The fact that 50-60% of cases with severe head injury result from traffic accidents underlines the great significance of emergency care and of its organization. Many patients with severe head injury are threatened from vital complications diagnosed with delay, or not at all, which plays a major role not only for survival but also for the quality of recovery and regaining of employment capabilities. Thus, the necessity of qualified and trained physicians with experience in emergency care is obvious. Emergency care can be divided into an early resuscitation phase of securing or reestablishment of general vital functions, and a following stabilisation phase with administration of measures directed towards the specific conditions underlying trauma. 1. Prevention and treatment of respiratory complications. In addition to classical emergency care measures, endotracheal suction might be employed. The most effective method for clearance of airways and, thus, securing of the cerebral oxygenation is endotracheal intubation. Early intubation provides also for control of the intracranial pressure by hyperventilation and administration of O2. Recently assistant ventilation is available as compared to the past when only controlled ventilation was possible. 2. Circulatory support. A major requirement for a sufficient cerebral perfusion is an adequate cerebral perfusion pressure making necessary early fluid substitution. In case the patient is in circulatory shock, shock-specific treatment may compete with adequate positioning of the patient. 3. Pharmacological treatment in the prehospital phase. Although dexamethasone has been reported to directly influence brain edema, its benefits in head injury are not clear. Currently conducted clinical studies using markedly higher doses may provide so far missing information.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421949 TI - Prehospital management of head injuries: international perspectives. AB - The extent of disability and, therefore, the management of head injured patients before their arrival to the hospital should be influenced by a number of factors including: the age of the victim (children with comparable severity of head injury have less disability and lower mortality than persons over the age of 40); the type of injury (motor-vehicle accidents are the most frequent cause of head trauma in the U.S. and Canada); the decision to transfer the patient to either the nearest hospital or to a designated neurotrauma center (at several communities in the U.S. and Canada, recent analysis has demonstrated that the extent of disability and the mortality among head injured victims can be significantly decreased by their admission to specialized trauma units); elapsed time as another important factor (at least in one type of traumatic injury--acute subdural hemorrhage--it has been shown than an interval of less than 2 hours between the time of injury and the time of the craniotomy can significantly decrease both the mortality and the extent of neurological impairment among the survivors). A number of acute injury effects on endothelial, neuronal or glial cells could potentially be influenced by compounds that may be administered before the victim arrives to the hospital.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421950 TI - Management of intracranial hypertension in head injury: matching treatment with cause. AB - Raised intracranial pressure (ICP) is common after head injury and strongly associated with mortality and morbidity. Empirical and prophylactic therapy with steroids and barbiturates has proved unsuccessful. Ideally, therapy should be targeted at the predominant cause of the increase in ICP. In head injury these may be (1) an increase in cerebral blood volume best treated by hyperventilation and hypnotic drugs. (2) an increase in brain water content best treated by osmotherapy and (3) increased CSF outflow resistance best treated by CSF drainage. This last cause seldom predominates in head injury. To determine whether it is possible to identify the best therapy in individual head injured patients, we are comparing osmotherapy (mannitol) and hypnotic drugs (thiopentone and gamma-hydroxybutyrate) in selected patients with severe head injury where it is possible to maintain standard conditions of ventilation and stable blood pressure and to measure ICP, CPP, brain electrical activity, PR ratio of the ICP wave form and cerebral AvDO2 before and during each of the two forms of therapy. Effective therapy means that ICP has been reduced to 20 mm Hg with preservation or improvement in CPP. 17 patients have been studied so far and 4 groups identified. Osmotherapy was superior to hypnotic in 5 cases, hypnotic superior to mannitol in 3 cases, both were effective in 5 cases and neither effective in 4 cases. Patients in whom hypnotics were superior tended to be younger, with diffuse rather than focal brain injury, had the highest levels of brain electrical activity prior to treatment and a higher PR ratio.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421951 TI - Traumatic brain tissue acidosis: experimental and clinical studies. AB - We have been focusing on potential metabolic derangement associated with severe head injury and a clinical trail directed toward treating brain tissue acidosis is currently underway. More specifically, we based this study on the hypothesis that following brain trauma brain tissue acidosis develops which may contribute to the prolongation of coma and neurologic deficit. Tromethamine (THAM), a safe and low toxicity agent which buffers in major part by causing a hypocapnic alkalosis, was selected for trial. Patients admitted with GCS < 8 were randomized into one of three arms: control: THAM plus hyperventilation; hyperventilation alone. Each regimen was maintained for 5 days post injury. Our analysis of 3 and 6 months Glasgow outcome score showed that prophylactic hyperventilation retards recovery, and the use of THAM overcomes the apparent deleterious effects of hyperventilation. One explanation is that the reduced ICP instability observed in THAM treated patients may account for this improvement. Is THAM effective in buffering traumatized brain tissue? What factors account for improvement in ICP stability? We addressed these questions in experimental studies utilizing MR spectroscopy to measure brain lactate production and tissue pH in fluid percussed anaesthetized cats. The protocol was designed to match our clinical trial, and brain injured animals were randomized into control, THAM, and hyperventilated groups. We observed that brain lactate production increased with trauma and remained above control at 8 hrs post injury. Lactate production in THAM treated animals was not elevated. Highest lactate production was associated with injured animals treated with sustained hyperventilation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421952 TI - NMR-spectroscopic investigation of cerebral reanimation after prolonged ischemia. AB - The severity of brain injury following interruption of blood flow depends on a number of ischemic and post-ischemic variables. The most important ischemic variables are the duration of ischemia, the amount of residual blood flow, the type and depth of anesthesia, brain glucose content and temperature. Among the post-ischemic factors the no-reflow phenomenon, edema and a variety of biochemical disturbances are of particular importance. Due to the complex interaction of these factors irreversible brain injury usually occurs after less than 10 min cerebrocirculatory arrest in normothermia. However, the safe ischemia time of the brain can be substantially extended when appropriate therapeutic measures are used to alleviate post-ischemic injury. NMR-spectroscopy is particularly suited for the analysis of this process. Recording of 31P, 1H and 19F spectra allow the continuous non-invasive assessment of such basic parameters as brain energy state, tissue pH, the content of lactate and blood flow (using Freon-23 as an inert tracer). In addition, information is obtained about changes in the content of phosphomonoesters and -diesters, glutamate, glutamine, aspartate and N-acetyl aspartate. These measurements can be combined with in vivo electrophysiological and post-mortem biochemical investigations for the further refinement of functional/metabolic monitoring. We have used this approach to study the potentials of post-ischemic resuscitation after one hour complete ischemia of the normothermic cat brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421953 TI - Quantification of primary and secondary lesions in severe head injury. AB - The current trend is to classify brain damage due to non-missile head injury as focal or diffuse. Quantitative findings will be discussed and reference will be made to both human and experimental non-human primate. PMID- 8421954 TI - Traumatic damage to the nodal axolemma: an early, secondary injury. AB - Electronmicroscopical investigations were made in a model of optic nerve damage in guinea-pigs on the development of acute axonal damage on an ultrastructural basis. It was expected to obtain thereby further information on mechanisms underlying axonal damage in traumatic brain injury. For that purpose an injury apparatus was employed to deliver defined elongation and/or tensile strains to the optic nerve. Transmission electronmicrographs were examined of longitudinal and transverse nerve sections throughout its entire length. The most severe abnormalities were identified in the prechiasmatic portion of the nerve. Among others, elongations of the nodes of Ranvier were encountered, swollen axons with accumulation of organelles, and even disrupted axons having a morphology similar to retraction balls. In all instances, abnormal axons were found together with axons having a normal structural appearance. Nodes of Ranvier demonstrated outward dilatations of the nodal axolemma and of the adjacent axoplasm, which are named as nodal blebs. Nodal blebs occurred already 15 min after injury, and were fully developed at 6 or 24 hrs. The blebs had disappeared again after 5-7 days. The axoplasm in the blebs demonstrated considerable disorganization of cytoskeletal elements with an array of amorphous material appearing as granular degeneration. Taken together, the present experimental model is a useful approach to analyse axonal damage at the ultrastructural level as it may occur in white matter of the central nervous system. PMID- 8421955 TI - Morphometrical evaluation of triflusal in brain infarction. AB - MCA occlusion in animals is a common model for experimental stroke. In previous studies we have shown that one of the factors, which influence evolution of an infarct is microthrombosis in the area of infarction and in the surrounding brain tissue. The present study was undertaken for assessment of the number of microthrombi and of the size of brain infarcting in rats treated with the antiaggregatory substance Triflusal. 7 groups of Sprague-Dawley rats, each group consisting of 6 animals, underwent transsphenoidal MCA occlusion. The animals received Triflusal in various amounts from day 2 till day 6. At day 7 animals were decapitated and the brains were fixed in formaldehyde. The brain was dissected at the level of the optic chiasm and embedded in paraffin. Fresh microthrombi were detected py PTAH (Phosphotungstic acid hematoxylin) staining. In each animal the hemisphere with the ischemic lesion as well as the contralateral hemisphere were examined. The area of both hemispheres was calculated by subtraction of the ventricle area from the total brain area of a section. Infarct was defined as the region of necrosis which was sharply demarcated from normal brain. The infarcted area was planimetrically measured to obtain a ratio of infarcted to normal brain. A correlation between the effect of Triflusal, number of microthrombi and size of the infarcted area could be demonstrated. The pathogenetic role of the microthrombi in the evolution of cerebral infarction as well as the effect of Triflusal in different dosages on the number of microthrombi could be clearly assessed by quantitative morphometry. PMID- 8421956 TI - In vivo and in vitro control of acid-base regulation of brain cells during ischemic and selective acidic exposure. AB - The three-compartment model of brain acid-base regulation postulates that under circumstances of changing function or disease, hydrogen ion concentrations may differ considerably in the interstitial space (ISS), the neurons and the glial cells. During hyperglycemia plus profound ischemia, for example, direct measurements by microelectrodes followed by intracellular HRP staining show that intraglial pH can fall transiently as low as 3.9, although more often the nadir drops to the 4.5-5.5 range. Concurrently, ISS-pH and, by calculation, neuronal pH fails to and remains constant (but not necessarily the same) at pH 6.2. By contrast, during spreading depression, ISS and intraglial pH at first move rapidly and transiently in opposite directions, ISS [H+] rising, intraglial falling. These two then gradually stabilize, whereas neuronal pH remains substantially more steady and near normal, shifting only minimally from resting baseline levels over several minutes' time. Similar but less pronounced effects follow direct electrical stimulation. The net change represents complex biophysical transmembrane and buffering mechanisms that appear to guard neuronal homeostasis. Studies carried out on embryonic rat forebrain neurons and glia show that these cells have considerably different vulnerabilities to extracellular acidity depending on the anionic nature of the acid in the bathing medium. In cultures to which HCI was added to the medium, neurons and neuronal processes almost all survived ten minute exposures to pH 3.8, whereas glial cells succumbed after ten minute exposures at pH not lower than 4.2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421957 TI - Glutamate receptor antagonists in experimental focal cerebral ischaemia. AB - Excessive activation of the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor has been implicated in the sequence of neurochemical events in cerebral ischaemia that results in irreversible neuronal damage. The effects of the NMDA antagonist MK-801 upon the amount of ischaemic brain damage has been assessed quantitatively in a cat and in a rat model of focal cerebral ischaemia. In chloralose-anaesthetised cats, focal cerebral ischaemia was produced by permanent occlusion of one middle cerebral artery (MCA) and the animal sacrificed 6 hours later. Pretreatment with the non-competitive NMDA antagonist, MK-801 (5 mg/kg, i.v.) reduced significantly the volume of ischaemic damage in the cerebral cortex by 57% compared to vehicle-treated cats. A similar degree of neuroprotection could be demonstrated in the cat MCA occlusion model if treatment with MK-801 was initiated 2 hours after the induction of ischaemia. In halothane-anaesthetised rats, focal cerebral ischaemia was produced by permanent MCA occlusion and the animals sacrificed 3 hours later. Pretreatment with MK-801 (0.5 mg/kg, i.v.) reduced the volume of ischaemic damage in the cerebral cortex by 38%; treatment with MK-801 initiated 30 minutes after MCA occlusion was equally effective in reducing cortical damage. In contrast to calcium entry blockers such as nimodipine in the rat MCA occlusion model, the improved histopathological outcome with MK-801 is not associated with improvement in cerebral tissue perfusion to the ischaemic tissue. The increasing evidence that NMDA receptor antagonists are beneficial in experimental focal cerebral ischaemia is reviewed. PMID- 8421958 TI - Effects of acute isotonic saline administration on serum osmolality, serum electrolytes, brain water content and intracranial pressure. AB - Albino rabbits who had undergone a cryogenic insult over the left parieto occipital cortex 24 hours previously were analyzed for serum osmolality, serum electrolytes, brain water content, intracranial pressure (ICP), following a 3 hour baseline intravenous infusion of above maintenance isotonic saline, and compared to sham operated controls. In the acute setting there was no difference in the serum osmolality and electrolytes between the subgroups. There was a significant increase in the water content of the white matter of the left hemisphere in the cold lesion group when compared to sham operated controls. Despite the intravenous fluid challenge, the ICP did not rise during the 3 hour experimental trial when compared to pre-trial values. It is concluded that in the acute setting an isotonic fluid load is compensated without significant disturbances of the above measured parameters, and should not alter therefore intracranial dynamics per se, in acute resuscitation measures following brain insults. PMID- 8421960 TI - Clinical features and classification of inherited ataxias. PMID- 8421959 TI - Cerebral metabolic studies in vivo by combined 1H/31P and 1H/13C NMR spectroscopic methods. AB - Intracellular pH and ammonium ion concentration are potent modulators of cerebral amino acid metabolism. Furthermore, intracellular acidosis and hyperammonemia accompany conditions such as ischemic encephalopathy and seizures and may contribute to the pathological sequelae observed. In vivo NMR spectroscopy permits multiple, non-destructive measurements of important cerebral metabolic intermediates in the same animal. We describe here the use of 1H, and 31P NMR spectroscopy to investigate the effects of acute changes in intracellular pH and ammonium ions on cerebral glutamate, glutamine, and lactate levels in vivo. We then show how 1H NMR can be used to indirectly follow the flow of 13C label from [1-13C] glucose into the cerebral glutamate pool, allowing us to measure cerebral TCA activity in normal and chronically hyperammonemic rats. Male Sprague-Dawley rats (160-210 gm), fasted 24-hours, were tracheotomized, paralyzed and ventilated on 30% O2/70% N2O. NMR spectroscopy was performed at a field strength of 8.4 Tesla using a Bruker AM-360 wide bore spectrometer. An elliptical surface-coil (8 x 12 mm) was double-tuned to either the 1H and 31P or 1H and 13C frequencies. After retraction of extracranial tissues, the coil was positioned over the skull 2 mm posterior to the bregma. Tail arteries and veins were cannulated allowing periodic measurements of PO2, pCO2, pH and glucose in arterial blood and intravenous infusions. Respiratory acidosis was induced in rats by the addition of CO2 to the ventilation gas mixture. Arterial pCO2 increased within 5 min from a pre-hypercarbic value of 36.4 +/- 6.1 mm Hg to 200-220 mm Hg and was maintained at this level for over 1 hour. Hypercarbia led to rapid cerebral acidification. Intracellular pH decreased from 7.18 +/- 0.08 (pre-hypercarbic period) to 6.68 +/ 0.06 (n = 4) at 10 min and remained stable throughout the NMR observation period. Glutamate decreased to 53 +/- 4% of control after 60 min of hypercarbia, while glutamine increased to 126 +/- 7% of control. Acute hyperammonemia was produced by a programmed intravenous infusion of 250 mM ammonium acetate, which rapidly raised and maintained the concentration of ammonium ions in the blood at approximately 500 microM. Shortly after the start of the infusion (10-20 min), the levels of glutamine and lactate rose continuously throughout the experiment, reaching levels of 170 +/- 25% and 260 +/- 60% of control, respectively (n = 12) after 50 min. Glutamate decreased during the same time interval to 80 +/- 4% of control (n = 12).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8421961 TI - Early onset ataxias in Tunisia. Intrafamilial heterogeneity. PMID- 8421962 TI - Genetic aspects of ataxia telangiectasia. PMID- 8421963 TI - Linkage studies in dominantly inherited ataxias. PMID- 8421964 TI - Epidemiology and clinical aspects of Machado-Joseph disease. PMID- 8421965 TI - The anatomy and physiology of cerebellar disease. PMID- 8421966 TI - Inherited prion diseases. PMID- 8421967 TI - Oligodendrocyte- and myelin-associated inhibitors of neurite growth in the adult nervous system. PMID- 8421968 TI - Cerebellar grafting in murine heredodegenerative ataxia. Current limitations for a therapeutic approach. PMID- 8421969 TI - Molecular analysis of the Friedreich's ataxia locus. PMID- 8421970 TI - Clinical and imaging correlations in inherited ataxias. PMID- 8421971 TI - Clinical and molecular genetic studies on autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS). PMID- 8421972 TI - Impact of treatment strategy on predischarge exercise test in the Thrombolysis in Myocardial Infarction (TIMI) II Trial. AB - Predischarge supine bicycle ergometry was used to assess persistent myocardial ischemia in postinfarction patients who received thrombolytic therapy and were randomized to an invasive versus conservative strategy in the Thrombolysis in Myocardial Infarction (TIMI) II trial. The frequency of ischemic responses in both strategies, and the 1-year prognostic importance of the different exercise test outcomes were examined. At 14 days, the percentage of patients with any adverse outcome (including death, presence of exercise-induced ST-segment depression, or inability to perform the exercise test) was 33.7% of 1,681 randomly assigned to the invasive strategy compared with 34.6% of 1,658 randomly assigned to the conservative strategy (p = 0.57). The 1-year mortality was greater in patients who did not perform the predischarge exercise test (7.7%) than in those who did (1.8%) (p < 0.001); the former were older, and a greater proportion were women, had a more frequent history of myocardial infarction, and more extensive coronary artery disease (p < 0.01 for each comparison). The 1-year mortality in patients with exercise-induced ST-segment depression or chest pain was only 1.4% (3 of 22) among those randomly assigned to the conservative strategy where coronary angiography and revascularization were recommended if the test result was abnormal (relative risk compared with those without ST-segment depression or chest pain 0.6; 99% confidence interval 0.1 to 2.9).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421973 TI - Transient myocardial ischemia after a first acute myocardial infarction and its relation to clinical characteristics, predischarge exercise testing and cardiac events at one-year follow-up. AB - The relation between early out-of-hospital ambulatory ST-segment monitoring, clinical characteristics, predischarge maximal exercise testing and cardiac events was determined in 123 consecutive men (age 55 +/- 8 years) with a first acute myocardial infarction (AMI). During 36 hours of ambulatory recording 11 +/- 5 days after AMI 23 patients (19%) had 123 ischemic episodes (group 1), whereas 100 patients demonstrated no ischemia (group 2). Exercise-induced ST-segment depression was more prevalent in group 1 (83%) than in group 2 (47%) (p < 0.005). Group 1 patients also had more severe ischemia as judged from a shorter exercise duration before significant ST-segment depression (5.5 +/- 2.4 vs 7.7 +/- 4.1 minutes; p < 0.03) and more pronounced ST-segment depression on exercise testing (4.1 +/- 2.6 vs 2.6 +/- 1.6 mm; p < 0.03). Furthermore, exercise test results revealed an impaired hemodynamic response in group 1 compared with group 2: systolic blood pressure at maximal work load 160 +/- 31 vs 176 +/- 28 mm Hg (p < 0.025) and systolic blood pressure increase during exercise 41 +/- 24 vs 56 +/- 22 mm Hg (p < 0.01). With-in 368 +/- 8 days of follow-up the frequency of cardiac events (cardiac death, nonfatal reinfarction, and severe angina including the need of revascularization) was 52% in group 1 compared with 22% in group 2 (p < 0.01). Exercise-induced ischemia did not predict an adverse outcome: event rate 30 vs 25% in patients without residual ischemia (p = NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421974 TI - Continuously updated 12-lead ST-segment recovery analysis for myocardial infarct artery patency assessment and its correlation with multiple simultaneous early angiographic observations. AB - Early angiography may not adequately subgroup patients with myocardial infarction if cyclic changes in coronary flow occur frequently. From a pilot experience using a new 12-lead ST-segment monitor, a continuously updated, self-referenced ST-recovery analysis method was developed to quantify both instantaneous recovery, as a noninvasive marker of patency, and cumulative ST recovery over time, as a marker of the speed, stability and duration of reperfusion. In 22 patients with acute infarction in whom 44 observations of unique angiographic patency were noted within 6 hours of presentation, serial patency assessments simultaneous with all angiographic observations predicted coronary occlusion with 90% sensitivity and 92% specificity. Of the 22 patients, 11 (50%) had multiple ST trend transitions suggesting cyclic changes in coronary flow before catheterization. Speed, stability and duration of ST-segment recovery were defined by the time to first 50% ST recovery, total number of ST-trend transitions and patent physiology index (percentage of monitoring period showing ST recovery), respectively. Subgrouped angiographically, the median (interquartile range) for cumulative ST parameters with patent (n = 8) versus occluded (n = 14) arteries were, respectively--time to 50% recovery, 1.57 (1.16, 1.70) versus 0.17 (-0.47, 0.32) hours; number of reelevation/recovery events, 1.5 (1, 3) versus 3 (1, 3); and patent physiology index, 52 (47, 59) versus 50 (5, 73). Thus, continuous ST-segment recovery analysis appears to predict simultaneous angiographic patency over serial assessments, whereas cumulative parameters appear to contain independent information, probably because of patency changes before or after angiography. PMID- 8421975 TI - Frequency and prognostic significance of secondary ventricular fibrillation complicating acute myocardial infarction. SPRINT Study Group. AB - The incidence of secondary ventricular fibrillation (VF) complicating acute myocardial infarction (AMI) was 2.4% in a large cohort of unselected patients with AMI (142 of 5,839). Secondary VF was more frequent in patients with recurrent AMI (4%) than in those with a first AMI (1.9%) (p < 0.01). The hospital course was more complicated and in-hospital mortality was significantly higher in patients with secondary VF than in those with the same clinical hemodynamic condition but without VF (56 vs 16%; p < 0.0001). Multivariate analyses confirmed secondary VF complicating AMI as an independent predictor of high in-hospital mortality, with an odds ratio of 7 (95% confidence interval 4.6-10.6). However, long-term mortality after discharge (mean follow-up 5.5 years) was not increased in patients with as compared with those without secondary VF (39 vs 42%). These findings were also true when patients receiving beta blockers and antiarrhythmic therapy were excluded from analysis. Thus, this life-threatening arrhythmia occurring during hospitalization is not a marker of recurrent susceptibility to VF or an indicator of increased mortality after discharge from the hospital. PMID- 8421976 TI - Comparison of thrombolytic therapy for acute myocardial infarction in patients aged < 35 and > 55 years. AB - There are differences in the risk factor profile and coronary anatomy of young patients who develop coronary artery disease compared with those of older ones. There is an absence of data in published reports regarding the response to thrombolytic therapy and the outcome of acute myocardial infarction in young patients. Sixty-two patients aged < 35 years (group 1) were compared with 58 aged > 55 years (group 2) who presented with acute myocardial infarction and were treated with intravenous streptokinase. Group 1 had a significantly higher incidence of smoking (p = 0.0009) and a lower incidence of diabetes mellitus (p = 0.002) than did group 2. Fifty-eight patients in group 1 and 40 in group 2 were studied by angiography at a similar time (5 to 6 days) after admission. Patients in group 1 had a better left ventricular ejection fraction (55 +/- 13% vs 49 +/- 13%; p = 0.03), but similar patency rates of the infarct vessel (74 vs 73%) compared with those of group 2. Group 1 also had a higher incidence of insignificant disease (22.5 vs 2.5%; p = 0.008) and a lower incidence of 3-vessel disease (10 vs 37.5%; p = 0.004). This suggests that there are differences in the risk factor profiles and coronary anatomy of young patients compared with those of older ones. Despite similar benefits from thrombolytic therapy in the form of a patient infarct vessel, there may be differences in the long-term outcome among these patients. PMID- 8421977 TI - Relation of apolipoprotein E phenotype to myocardial infarction and mortality from coronary artery disease. AB - The apolipoprotein E polymorphism is a genetic determinant of low-density lipoprotein (LDL) cholesterol. Its status as a risk factor for coronary artery disease (CAD), either through a causal relation with LDL cholesterol level or independently, is less clearly established. Data from the Multiple Risk Factor Intervention Trial were used to examine the influence of apolipoprotein E phenotype on risk of coronary events. Of the 12,866 randomized participants, 619 were studied in a nested case-control design. CAD deaths (93) and nonfatal myocardial infarctions (113) were matched to 412 controls. The allele frequencies of apolipoprotein E in the white subset (epsilon 2 = 0.06, epsilon 3 = 0.79, and epsilon 4 = 0.15) were very similar to other nonselected white American populations, and the relation of apolipoprotein E on total and LDL cholesterol was generally similar to that seen in other studies, with the epsilon 2 allele being associated with lower and the epsilon 4 allele with higher total and LDL cholesterol. Allele frequencies were not the same for patients and control subjects. The presence of epsilon 4 was associated with an increased risk of CAD that was most evident for fatal cases. There was no relation between changes in LDL cholesterol over time during the trial and apolipoprotein E phenotypes. PMID- 8421978 TI - Value of rest thallium-201/technetium-99m sestamibi scans and dobutamine echocardiography for detecting myocardial viability. AB - The relation between radioisotopic and echocardiographic markers of myocardial viability and postrevascularization recovery of function is still to be defined. To this purpose, 14 patients (11 men, 3 women, aged 35 to 64 years, mean 54 +/- 7) with ventricular dysfunction were studied by a multiparametric approach. Each patient underwent, on separate days, rest thallium-201 and technetium-99m sestamibi scintigraphy, dobutamine echocardiography and coronary angiography. Coronary angiography was analyzed by a quantitative approach. Thallium uptake at rest was quantified from planar early (10-minute) and delayed (16-hour) thallium 201 images and expressed as a percentage of maximal activity in each projection using a 13-segment model. Sestamibi uptake was expressed in the same way. Dobutamine (up to 10 micrograms/kg/min) echocardiography was analyzed using a score index ranging from 1 (normokinesia) to 4 (dyskinesia) and a similar segmental model. Before revascularization 50 segments were grouped as normal (coronary stenosis < 50% and normal function, group 1); of the remaining 132 segments with > 50% coronary stenosis, 57 had normal wall motion (group 2) and 75 showed regional dyssynergies (group 3). Early and delayed thallium-201 regional percent activities did not differ in group 1 and in group 2 but were significantly less in group 3 segments. Sestamibi percent activity was more in group 1 and significantly reduced both in group 2 and 3 segments. Segments with improved wall motion after dobutamine had more early, delayed thallium-201 and sestamibi percent activities than unresponsive segments. Postrevascularization echocardiography was performed in all patients. Delayed thallium-201 scans and dobutamine echocardiography showed good sensitivity and specificity in detecting viable myocardium. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421979 TI - Unrecognized left main coronary artery disease in patients undergoing interventional procedures. AB - Selective, coronary arteriographic, catheter-based, intravascular ultrasound images were obtained to determine the presence and extent of angiographically undetected or underestimated left main (LM) coronary arterial narrowing in patients receiving coronary interventional therapy. Coronary arteriograms were determined to be either normal or abnormal by visual inspection. Abnormal arteriograms were digitized and quantitated using a semiautomated edge-detection algorithm. Thirty-eight patients receiving percutaneous treatment of stenoses in the left coronary artery system were studied. Optimal LM coronary angiograms were obtained in 2 views, and intravascular ultrasound images were obtained after the coronary interventional procedure. Intravascular ultrasound detected plaque in 24 of 27 angiographically normal LM arteries (89%), whereas narrowing was observed in 11 of 11 angiographically abnormal LM arteries (100%). Eight of 38 patients (21%) had > 40% area stenosis by intravascular ultrasound. In patients with angiographic disease, there was no correlation between quantitative angiographic and ultrasound percent area stenosis (r = 0.12; p = 0.72; SEE 19%). The median plaque area was not different between angiographically normal (0.05 cm2; 0.03, 0.08 [25th, 75th percentile]) and abnormal (0.06 cm2; 0.03, 0.1) patients. The median percent area stenosis in arteriographically normal subjects (26%; 14, 32%) was less than that in abnormal ones (37%; 20, 46%) (p = 0.03). Unrecognized LM disease is widespread and often underestimated in patients with normal LM angiograms undergoing interventional procedures. Plaque area is similar for angiographically normal and insignificantly abnormal vessels. This study suggests that intravascular ultrasound overcomes the limitations of silhouette imaging and can be a clinically useful, adjunctive method to evaluate LM coronary artery disease. PMID- 8421980 TI - Transient myocardial ischemia during nifedipine therapy in stable angina pectoris, and its relation to coronary collateral flow and comparison with metoprolol. AB - There are conflicting results concerning the anti-ischemic effect of nifedipine in patients with chronic stable angina. Therefore, the purpose of this study was to assess whether the anti-ischemic effect of nifedipine may be related to coronary collateral circulation. Forty-one patients with stable angina and coronary artery disease were randomized to a parallel double-blind study with nifedipine and metoprolol, and compared for effects on transient ischemic episodes during ambulatory electrocardiographic monitoring and exercise-induced ischemia. The effects were correlated to the presence of collateral circulation. In 17 patients, angiographically poor or no collateral flow was observed (group 1), and 24 had good collateral flow (group 2). Nifedipine was administered to 20 patients (8 in group 1, and 12 in group 2). In group 1, nifedipine reduced the frequency of total and asymptomatic ischemic episodes (p < 0.05), whereas significant increases in both total (p < 0.05) and silent (p < 0.01) ischemia were observed in group 2. Exercise variables were slightly improved (p = NS) during nifedipine therapy in group 1, and slightly worsened (p = NS) in group 2. Reflex tachycardia was not observed at either the onset of transient ischemia out of the hospital or exercise-induced ischemia. This was in contrast with the effect in 21 patients treated with metoprolol (9 in group 1, and 12 in group 2) where significant reductions were observed in the frequency of both total (p < 0.01) and silent (p < 0.01) ischemia in both groups. Furthermore, a beneficial effect was observed on all exercise variables.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421981 TI - Comparison of two mechanical ultrasound devices with and without echogenic contrast dye. AB - To evaluate the impact and limitations of intracoronary ultrasound, 64 segments of 13 isolated coronary arteries were examined and 2 mechanical devices (device A, 30 MHz 5Fr [CVIS] and device B, 20 MHz 4.8Fr [Diasonics/Boston Scientific] were compared with the corresponding histologic specimens. Luminal dimensions were assessed with and without echogenic contrast dye (Laevovist, Schering AG, Germany). After application of contrast dye, correlation of luminal area between histology and ultrasound was improved from r = 0.62 to r = 0.77 (device B; p < 0.05) and from r = 0.82 to r = 0.88 (device A; p = NS). Low accuracy of lumen measurements in segments < 2.5 mm could be improved by application of contrast dye. The number of quadrants in which wall thickness measurements were impossible was significantly higher for device B (n = 56; 22%) than for device A (n = 28; 11%); p < 0.01. This may be due to the different ringdown diameters of both systems (B, 2.6 mm; A, 2.0 mm; p < 0.0001). In assessing wall thickness only in segments of > 2.5 mm, a reliable correlation between ultrasound and histology was found (A, r = 0.80; B, r = 0.60). Sensitivity of plaque (n = 51) detection was lower for device B (63%) than for device A (82%, p < 0.05), and measurements correlated with histology only for device A. There are considerable differences in the accuracy of ultrasound measurements between mechanical systems. Nevertheless, additional application of contrast dye can improve accuracy of luminal measurements, especially in smaller vessels. PMID- 8421982 TI - Left atrial appendage flow velocity assessment using transesophageal echocardiography in nonrheumatic atrial fibrillation and systemic embolism. AB - Fifty-four patients with nonrheumatic atrial fibrillation (AF) were studied: 16 patients with (group I) and 38 patients without (group II) documented systemic embolism. Transesophageal echocardiography (TEE) was performed to evaluate the presence of left atrial (LA) appendage thrombus and LA spontaneous contrast, LA size, systolic and diastolic peak velocity of the left pulmonary vein, and forward and backward peak velocity of the LA appendage. No difference was observed in the presence of LA thrombus between the 2 groups. The occurrence of LA spontaneous contrast was significantly (p = 0.01) higher in the group with embolism. LA size, measured by atrial length (4.96 +/- 0.84 vs 4.79 +/- 1.38 cm; p = NS) and atrial width (4.50 +/- 0.96 vs 4.31 +/- 1.24 cm; p = NS), was the same for both groups and thus not associated with embolism. There was no difference in systolic peak velocity (0.39 +/- 0.22 vs 0.44 +/- 0.22 m/s; p = NS), and a trend toward a higher diastolic peak velocity (0.50 +/- 0.17 vs 0.42 +/- 0.15 m/s; p = 0.08) was seen in the left pulmonary vein in the group with embolism. Forward (0.25 +/- 0.19 vs 0.39 +/- 0.23 m/s; p < 0.05) and backward (0.23 +/- 0.15 vs 0.33 +/- 0.16 m/s; p < 0.05) peak velocities of the LA appendage were significantly lower in the embolism group. Assessment of LA appendage flow velocity may potentially identify patients with nonrheumatic AF at high risk for systemic embolism. PMID- 8421983 TI - Comparison of biphasic and monophasic shocks for defibrillation using a nonthoracotomy system. AB - A comparison of defibrillation thresholds was made using biphasic and monophasic shocks delivered by a nonthoracotomy lead system in 2 clinically distinct groups of patients. The first group were patients receiving an implantable cardioverter defibrillator who were studied before surgery with their chests closed. The second group were patients undergoing coronary artery bypass grafting (CABG) who were studied before surgery with their chests open but reapproximated. Biphasic defibrillation thresholds (stored energy) were significantly (p < 0.001) less than monophasic ones in subjects with the implantable cardioverter-defibrillator (12.3 +/- 5.3 vs 21.1 +/- 9.3 J) or CABG (14.6 +/- 7.1 vs 24.2 +/- 12.6 J). These values are less than were previously reported with a similar nonthoracotomy lead configuration. There were no significant differences between the 2 groups in all measurements derived from corresponding shock waveforms, although impedance tended to be greater in patients with CABG. However, subjects with CABG had greater left ventricular ejection fractions and did not have history of potentially lethal ventricular arrhythmias. Despite these differences, the conclusion that biphasic shocks are more effective would have been made in a study of either group alone. It is concluded that patients with CABG who have not had preceding potentially lethal ventricular arrhythmias may be a potential source of surrogate subjects for defibrillation research such as epicardial mapping, which requires that the chest be open. PMID- 8421984 TI - Effects of intravenous milrinone on left ventricular function in ischemic and idiopathic dilated cardiomyopathy. AB - M-mode echocardiography and Doppler were used to assess the effects of phosphodiesterase inhibition on subendocardial function in dilated cardiomyopathy, and in particular, to study interactions with both systolic and diastolic left ventricular function. Twelve adult patients with dilated cardiomyopathy were studied (6 ischemic in origin and 6 idiopathic), 7 of whom were being considered for cardiac transplantation. Cardiac index increased without significant change in heart rate or blood pressure. Longitudinal mitral ring motion, which had been uniformly reduced, increased markedly after intravenous milrinone. Left ventricular cavity size decreased, and shortening fraction, posterior wall thickness, and rates of posterior wall thickening and thinning increased markedly. Left atrial pressure decreased, and isovolumic relaxation time increased. However, the peak velocity and duration of the transmitral E wave increased, with no change in the A wave. Improved longitudinal (subendocardial) function was reflected by improved posterior wall dynamics, and early filling, possibly by augmentation of restoring forces. Thus, severely reduced subendocardial function in dilated cardiomyopathy is potentially reversible, with marked effects on systolic and diastolic function. These previously unrecognized actions of milrinone provide further evidence to justify its short-term use in supporting the severely depressed myocardium. PMID- 8421985 TI - Comparison of transthoracic and transesophageal echocardiography for detection of abnormalities of prosthetic and bioprosthetic valves in the mitral and aortic positions. AB - Two-dimensional echocardiography is the diagnostic procedure of choice for evaluation of prosthetic valve abnormalities. However, transthoracic echocardiography (TTE) may be limited owing to acoustic shadowing and poor acoustic windows. Some of these limitations may be overcome by transesophageal echocardiography (TEE). One hundred twenty-six patients with 148 prosthetic valves (113 bioprostheses and 35 mechanical devices) were studied by M-mode and 2 dimensional TTE and TEE. Prosthetic valve morphology was confirmed by surgery or autopsy in all cases; 124 prostheses were classified as diseased (33 endocarditis, 8 thrombi, and 83 degeneration defined as leaflet thickening > 3 mm with restricted motion) and 24 as normal. Prosthetic valve endocarditis and thrombi were correctly identified by TTE in 12 of 33 (36%) and 1 of 8 (13%) prostheses, respectively, but could be diagnosed by TEE in 27 of 33 (82%; p < 0.001) and 8 of 8 (100%; p < 0.01), respectively. Compared with TTE, TEE had a higher sensitivity for morphologic prosthetic valve abnormalities in patients with either bioprostheses (88 [87%] vs 66 [65%] of 101 prostheses; p < 0.01) or mechanical devices (19 [83%] vs 5 [22%] of 23 prostheses; p < 0.01) and in patients with a prosthesis in either the aortic (49 [77%] vs 32 [50%] of 64; p < 0.01) or mitral (58 [97%] vs 39 [65%] of 60; p < 0.001) position. Overall, sensitivity and specificity were 57 and 63%, respectively, for TTE, and 86 and 88%, respectively, for TEE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421986 TI - Color Doppler echocardiographic determination of mitral regurgitant flow from the proximal velocity profile of the flow convergence region. AB - Flow rate across an orifice can be determined from color Doppler echocardiographic maps of the flow convergence region proximal to the orifice. Different methods have been developed in vitro. The proximal velocity profile method was prospectively evaluated in patients with mitral regurgitation. Color Doppler echocardiography was performed in 74 patients before cardiac catheterization. The increasing velocities within the flow convergence region were determined in an apical plane on the straight line from the transducer to the leak; thus the proximal velocity profile was established and plotted on a nomogram. Instantaneous regurgitant flow rate was derived from the position of the resulting curve in relation to the nomogram's reference curves, which were derived from in vitro measurements. Regurgitant stroke volume was calculated as regurgitant flow rate.regurgitant velocity-time integral/regurgitant peak velocity, using additional continuous-wave Doppler. The 55 patients with angiographic regurgitation had a close association between regurgitant flow rate (0 to 600 ml/s) and angiographic grade (Spearman's rank correlation coefficient = 0.91; p < 0.0001). Regurgitant flow rate did not overlap between grades < or = 2+, 3+ and 4+. In 16 patients, regurgitant stroke volume by echocardiography correlated well with that by the angiography/Fick method (r = 0.88; SEE = 17.1 ml), with a regression line close to identity (y = 0.89x + 12.7 ml). The proximal velocity profile method enables determination of mitral regurgitant flow and estimation of regurgitant volume. PMID- 8421987 TI - Cineradiography for determination of normal and abnormal function in mechanical heart valves. AB - To determine the diagnostic value of cineradiography of mechanical heart valves, 112 cinefluoroscopic studies were performed in 76 patients with 95 valve prostheses (caged ball or disk valves, tilting disk and bileaflet valves). A patient group (n = 45) presenting with clinical or echocardiographic findings suggestive of valve-related complications was compared with a control group (n = 31) without such symptoms. Disk-opening angles (mean +/- SD) for Medtronic Hall aortic valves were found to be significantly smaller (62.8 +/- 11.1 degrees) in patients than in control subjects (73.9 +/- 1.6 degrees; p < 0.05). Tissue ingrowth or thrombus formation, or both, demonstrated in 3 patients on subsequent reoperation, are considered as the main cause of incomplete or asymmetric disk opening. Opening and closing times did not differ significantly between patients and control subjects. Besides abnormal valve motion, structural defects such as strut fracture or leaflet escape could be rapidly detected by cineradiography if x-ray projections according to the particular valve design were used. Together with quantitative Doppler echocardiographic and clinical data, this method can help to give specific answers if the question is to either confirm or exclude imminent or acute valve malfunction. Thus, modern cineradiography is a highly valuable noninvasive diagnostic tool for both rapid management of emergency cases and routine follow-up of patients with mechanical heart valves. PMID- 8421988 TI - Balloon mitral commissurotomy in patients aged > or = 70 years. AB - Of 280 patients treated by balloon mitral commissurotomy (BMC) between 1987 and 1991, 28 (10%) were > or = 70 years old. Two patients with associated significant aortic stenosis were excluded from the study. Older patients more often were in New York Heart Association class III or IV (84 vs 67%; p < 0.007) and atrial fibrillation (61 vs 36%; p < 0.0001), and had a higher echocardiographic score (9.3 +/- 2 vs 8 +/- 1.6; p < 0.0004) and a lower baseline cardiac index (2.1 +/- 0.6 vs 2.4 +/- 0.6 liters/min/m2; p < 0.03) than younger ones. Baseline mean pulmonary pressure (37 +/- 11 vs 34 +/- 12 mm Hg), transmitral gradient (14 +/- 4 vs 14 +/- 5 mm Hg) and valve area (1.0 +/- 0.4 vs 1.1 +/- 0.3 cm2) were not different between older and younger patients (p = NS). Acute complications during the procedure (including cardiac perforation, embolism, severe mitral regurgitation and surgical atrial shunt), and 30-day mortality after BMC were more frequent in older than younger patients (27 vs 9% [p < 0.01], and 12 vs 0.8% [p < 0.005], respectively). A complete success, defined as a mitral valve area increase > 25% and postmitral valve area > 1.5 cm2 was obtained in 16 of the 22 older patients (72%) with the completed procedure (compared with 81% of younger ones; p = 0.1).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8421989 TI - Effects of successful cardiac transplantation on plasma endothelin. AB - After cardiac transplantation, cyclosporine-treated patients exhibit a high incidence of systemic hypertension, the mechanism of which is not known. Endothelin, a potent vasoconstrictor peptide of endothelial origin, may be activated by cyclosporine-induced endothelial injury and therefore may mediate post-transplant hypertension. In the present study, we tested whether immunoreactive endothelin-1 could be detected by radioimmunoassay in the plasma of cardiac transplant recipients and if levels correlated with hemodynamic characteristics, cyclosporine level, or renal function as assessed by serum creatinine. Plasma endothelin was measured in 22 stable cyclosporine-treated patients 9 days to 3 years after successful orthotopic cardiac transplantation before routine hemodynamic assessment and surveillance endomyocardial biopsy. Fifteen patients were receiving chronic therapy for hypertension. Plasma endothelin-1 was 5.2 +/- 1.8 pg/ml (range 3.1 to 10.5), which was increased compared with that in 12 normal subjects (1.9 +/- 0.3 pg/ml; range 1.4 to 2.4); the difference was statistically significant (p < 0.0001). Repeated sampling in 8 patients at weekly intervals identified a persistent increase in endothelin with only modest variability. Endothelin-1 did not correlate with any hemodynamic variable, serum creatinine or cyclosporine level. Thus, endothelin-1 is increased after successful orthotopic cardiac transplantation. In the absence of discrete correlations with hemodynamic variables, serum creatinine or cyclosporine levels, both the characteristics and mechanisms for increased endothelin in recipients of cardiac transplants require further evaluation. PMID- 8421990 TI - Gender independence of ambulatory electrocardiographic monitoring in predicting perioperative cardiac risk. PMID- 8421991 TI - Acute pulmonary edema after Tityus serrulatus scorpion sting in children. PMID- 8421992 TI - Effects of coronary angioplasty using progressive dilation on ostial stenosis of the left anterior descending artery. PMID- 8421993 TI - Effects of adenosine on atrial repolarization in the transplanted human heart. PMID- 8421994 TI - Atrial flutter causes pseudo late potentials on signal-averaged electrocardiogram. PMID- 8421995 TI - Signal-averaged electrocardiography in asymptomatic alcoholics. PMID- 8421996 TI - Balloon angioplasty of aortic recoarctation. PMID- 8421997 TI - Thrombolytic therapy after cardiopulmonary resuscitation in acute myocardial infarction. PMID- 8421998 TI - Antifibrotic effects of spironolactone in preventing myocardial fibrosis in systemic arterial hypertension. AB - Activation of the renin-angiotensin-aldosterone system in arterial hypertension can lead to remodeling of the myocardial collagen network, with progressive collagen accumulation in the cardiac interstitium. This reactive myocardial fibrosis, which is not secondary to myocyte necrosis, appears to be an important determinant of diastolic dysfunction and thus of pathologic hypertrophy. To examine the effects of the aldosterone antagonist spironolactone on myocardial fibrosis, we analyzed interstitial fibrosis in 7 different models of arterial hypertension in rats: 2 kidney, 1 clip model of renovascular hypertension (RHT); continuous subcutaneous aldosterone (0.75 micrograms/hr) infusion; RHT and aldosterone models treated with 20 mg/kg per day of subcutaneous spironolactone; uninephrectomized rats on a high sodium diet; and age- and sex-matched controls with or without spironolactone treatment. Systolic arterial pressure was comparably elevated in RHT and aldosterone models; it was modestly lowered but not normalized by 8 weeks of spironolactone treatment at the low doses used. Such treatment also failed to prevent left ventricular hypertrophy (LVH) in all experimental groups with hypertension. Spironolactone, however, was able to prevent myocardial fibrosis in RHT and aldosterone models of acquired arterial hypertension irrespective of the development of LVH and the presence of hypertension. These findings provide further evidence that elevated aldosterone levels play an important role in the adverse remodeling of the myocardium in arterial hypertension. The antifibrotic effects of spironolactone, if confirmed in human studies, may be a valuable strategy in treating hypertensive heart disease. PMID- 8421999 TI - Influence of arterial blood pressure and aldosterone on left ventricular hypertrophy in moderate essential hypertension. AB - In a group of 36 untreated patients with mild-to-moderate essential hypertension (office systolic blood pressure [SBP] 160 +/- 3.4 mm Hg, office diastolic blood pressure [DBP], 102 +/- 1.5 mm Hg), 24-hour ambulatory blood pressure monitoring, and determination of left ventricular (LV) mass index according to the formula of Devereux were performed. After an overnight fast, blood samples were taken for the determination of serum aldosterone levels and plasma renin activity. Urinary catecholamine concentrations were assayed from 24-hour urine collections. Left ventricular mass index (143.7 +/- 8 g/m2) did not correlate significantly with either office SBP or office DBP. The correlation of LV mass index with mean 24 hour SBP (145 +/- 3 mm Hg) was statistically significant: r = 0.395, p = 0.026. However, the best correlation was obtained with mean 24-hour DBP (90 +/- 3 mm Hg) with r = 0.499 (p = 0.004). Urinary catecholamine levels did not correlate with LV mass index. In addition, LV mass index correlated significantly with plasma renin activity (r = 0.346, p = 0.050) and serum aldosterone levels (r = 0.559, p = 0.0009). There was a strongly significant correlation between LV mass index and serum aldosterone levels even after adjustment for mean 24-hour SBP (r = 0.496, p = 0.005) and DBP (r = 0.514, p = 0.004). These results demonstrate that ambulatory blood pressure determinations but not office blood pressure parameters correlate well with left ventricular hypertrophy in essential hypertension. Nonhemodynamic factors are important determinants of left ventricular mass as well.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422000 TI - Spironolactone in congestive heart failure refractory to high-dose loop diuretic and low-dose angiotensin-converting enzyme inhibitor. AB - Patients with severe congestive heart failure (New York Heart Association [NYHA] functional classes III-IV) often can tolerate only low doses of angiotensin converting enzyme (ACE) inhibitors because pronounced hypotension caused by additional ACE inhibitor increments may decrease renal perfusion. The use of high dose loop diuretics is currently advised to overcome diuretic resistance in refractory congestive heart failure (CHF). In a baseline controlled study, we evaluated 21 patients with diuretic resistance and evident fluid retention for the responses to 5 days of double drug therapy consisting of high-dose loop diuretic (10 mg oral bumetanide) in combination with the maximum tolerable dose of an ACE inhibitor (individualized to blood pressure and kidney function). Five patients (24%) showed a gross natriuresis and reduction in excess weight > 25% in response to this therapy. The remaining 16 patients (76%) with insufficient responses (i.e., < 25% reduction in excess weight) subsequently received 100 mg spironolactone once a day for 7 days in addition to the double therapy. Spironolactone coadministration was highly effective in 13 of 16 patients (81%). Marked natriuresis and diuresis were achieved within the next week of treatment, and CHF symptoms regressed or disappeared. The clinical course was similar in the bumetanide-ACE inhibitor and the bumetanide-ACE inhibitor-spironolactone treatment (triple therapy) groups. Plasma aldosterone was significantly higher (p < 0.05) in the patients who needed spironolactone. The 3 patients who were considered refractory to triple therapy exhibited the highest baseline plasma aldosterone concentrations.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422001 TI - Captopril and spironolactone therapy for refractory congestive heart failure. AB - Short- and long-term clinical effects of the angiotensin-converting enzyme (ACE) inhibitor captopril in severe congestive heart failure (CHF) were evaluated during a 3-year open study of 124 inpatients with New York Heart Association (NYHA) functional class III or IV CHF refractory to treatment with cardiac glycosides and high doses of loop diuretics. Captopril was added to each patient's regimen, which comprised combinations of furosemide (124 patients), digitalis (117 patients), and spironolactone (90 patients). By the end of the first month of captopril administration, improvement in NYHA functional class was seen in 89 patients (72%). During the first year of captopril treatment, the number of hospital admissions and hospital days declined significantly (p < 0.001) and functional class improved significantly (p < 0.001). Although most patients tolerated captopril well, 44% experienced hypotension, which in 10% of patients necessitated termination of captopril therapy. Although mean serum potassium levels tended to increase, serious hyperkalemia did not occur. After 1 year, a subset of 30 patients who had not initially received spironolactone deteriorated clinically and manifested increasing urinary aldosterone levels. Hypotension precluded increasing the captopril dose, but introduction of spironolactone improved clinical status in this cohort. The results suggest that rational therapy for severe CHF includes addition of the aldosterone antagonist spironolactone to low doses of captopril (or another ACE inhibitor) and high doses of loop diuretics, provided renal function is adequate. PMID- 8422002 TI - Aldosterone and antialdosterone therapy in congestive heart failure. AB - The pathophysiologic cycle that links myocardial failure with the appearance of congestive heart failure is not fully understood. It is clear, however, that an activation of several neurohormonal systems and the interplay between kidneys, adrenal glands, and heart contribute to abnormal sodium and water homeostasis. Aldosterone, the body's most potent mineralocorticoid hormone, contributes to intravascular and extravascular volume expansion, and thus to the appearance of symptomatic failure. Antialdosterone therapy in patients with secondary hyperaldosteronism due to heart failure must achieve one or more of the following goals: reduce or, preferably, normalize plasma aldosterone levels by limiting synthesis; antagonize the renal and systemic effects of aldosterone at its receptor sites; and eliminate or minimize the multiple stimuli to aldosterone secretion. PMID- 8422003 TI - Angiotensin-converting enzyme inhibitor and spironolactone combination therapy. New objectives in congestive heart failure treatment. AB - Secondary aldosteronism has deleterious effects in patients with congestive heart failure (CHF) and can contribute to congestion, ventricular arrhythmias, and sudden death. Mortality is higher in patients with elevated levels of plasma aldosterone. Aldosterone increases as CHF progresses as a result of activation of the renin-angiotensin-aldosterone system (RAAS). This is further amplified by the routine use of diuretics. Angiotensin-converting enzyme (ACE) inhibitors produce a profound and consistent inhibition of angiotensin II production, but they exert only a mild and transient antialdosterone effect. In a number of studies involving ACE inhibitors, plasma aldosterone levels at the end of the trial do not differ significantly from baseline. Spironolactone, a specific aldosterone receptor antagonist, may exert an independent and additive effect to that of ACE inhibitors. Apart from its renal effects, recent evidence suggests that spironolactone may exert direct cardiac and vascular effects inhibiting cardiac collagen hypertrophy and limiting vascular constriction. Combining an ACE inhibitor and spironolactone may achieve a more complete inhibition of the whole RAAS and may produce further clinical benefits. The efficacy and safety of such a combination has not been properly addressed. In the CONSENSUS trial, plasma potassium and creatinine levels were not necessarily adversely affected when enalapril was added to the regimens of patients receiving spironolactone, a condition existing in > 40% of the patients enrolled in this study. One prospective open study and other anecdotal reports suggest that combining spironolactone and ACE inhibitors resulted in clinical improvement without serious side effects in patients who could not tolerate further increases in the ACE inhibitor dose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422004 TI - Dose-related cardiovascular effects of spironolactone. AB - The aim of this study was to assess the short-term hemodynamic effects of increasing doses of spironolactone (25, 50, and 75 mg/day) on oliguric patients (5 men, mean age 47 +/- 12 years) undergoing hemodialysis for chronic renal impairment. Parameters of interest included heart rate (HR), cardiac output, systemic vascular resistance (SVR), arterial pressure, right atrial pressure, and pulmonary capillary wedge pressure (PCWP). The study also evaluated how spironolactone modified the effects on arterial and right atrial pressures and PCWP of infusion of increasing doses of norepinephrine (20, 40, and 100 ng/kg/min) and angiotensin II (2, 4, and 10 ng/kg/min). Compared with placebo, the lowest dose of spironolactone (25 mg/day) produced statistically significant (p < 0.05) modifications in systemic arterial pressures without a compensatory increase in cardiac output. The modifications were more pronounced at 50 and 75 mg/day, and the latter had a significant dose-dependent effect. Moreover, doses of 50 and 75 mg/day produced significant (p < 0.05) decreases in right atrial pressure and PCWP. Spironolactone administration caused the curve expressing the relation between an infused norepinephrine or angiotensin II dose and the blood pressure response to shift significantly (p < 0.05 to < 0.01) to the right, and the pressor doses of norepinephrine or angiotensin II showed a significant (p < 0.05 to < 0.01) dose-related increase, suggesting that treatment with spironolactone inhibited cardiovascular reactivity. This effect was observed on both the capacitance (i.e., low-pressure) and resistance (i.e., high pressure) vessels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422005 TI - Role of mechanical and hormonal factors in cardiac remodeling and the biologic limits of myocardial adaptation. AB - Patients with chronic congestive heart failure manifest > or = 1 of the following abnormalities: diastolic dysfunction, systolic dysfunction, and arrhythmias. Diastolic dysfunction, one of the first symptoms to occur during hypertensive cardiopathy, depends on both active relaxation of the cardiac muscle and passive ventricular compliance. The ability of the ventricles to relax depends on normal calcium metabolism and adenosine triphosphate concentration. Ability to extrude intracellular calcium is depressed in the hypertrophied, overloaded heart as compared with the normal myocardium. Myocardial fibrosis is the major cause of increased diastolic ventricular stiffness. Left ventricular (LV) hypertrophy and myocardial fibrosis also greatly increase the likelihood of ventricular arrhythmias, in particular by prolonging the QRS interval and facilitating the occurrence of reentry arrhythmias. Findings in animal studies have indicated that such fibrosis, which involves excessive collagen deposition, is independent of LV hypertrophy and that LV hypertrophy does not necessarily result in myocardial fibrosis. Instead, the development of myocardial fibrosis is sensitive to circulating levels of both angiotensin II and aldosterone, and the fibrotic response to each of these substances is independent. The aldosterone antagonist spironolactone prevents myocardial fibrosis in several animal models, thus confirming the importance of aldosterone in the genesis of excessive collagen deposition. PMID- 8422006 TI - Postmortem diagnosis of acute anaphylaxis by serum tryptase analysis. A case report. AB - Systemic anaphylaxis is an acute allergic emergency resulting from generalized mast cell degranulation. In the United States, it is estimated that anaphylaxis accounts for about 500 deaths each year. Hymenoptera-sting hypersensitivity is one of the most common causes of systemic anaphylaxis. The authors report a case of a healthy 26-year-old man who developed acute anaphylaxis after a bee sting, could not be resuscitated, and died within 1 hour. At autopsy, performed 14 hours after the event, the only pathologic findings were laryngeal edema and congestion of lung. Postmortem tryptase levels in the blood were obtained and were instrumental in confirming a diagnosis of acute anaphylaxis. This case is reported to discuss the difficulties associated with using traditional histamine levels in making a diagnosis of anaphylaxis and to validate the value of using tryptase levels to document acute anaphylaxis as a cause of death, even when serum is not obtained until many hours after death. PMID- 8422007 TI - Isolation of Borrelia burgdorferi from skin biopsy specimens of patients with erythema migrans. AB - Procedures for the cultural isolation and identification of Borrelia burgdorferi from skin biopsy specimens are described. B. burgdorferi was isolated from 24 of 34 skin biopsy specimens from patients with erythema migrans. Eight of the culture-positive patients had single, primary lesions and 16 had multiple, secondary lesions. The 17 male and 7 female patients were 2 to 70 years old. Biopsy samples were obtained from erythematous or normal-appearing skin within 1 cm of the peripheral aspect of the lesion. Twenty-three of the isolates were detected within 8 days of incubation in Barbour-Stoenner-Kelly medium with no antimicrobial agents. The identities of the isolates were determined by reactivity with monoclonal antibodies H9724 and H5332. Cultivation of B. burgdorferi from skin lesions suggestive of erythema migrans is a practical and clinically relevant procedure. Clinical isolates and corresponding patient sera and urine will contribute to efforts to improve existing immunoserologic testing methods and develop new assays to diagnose Lyme borreliosis. PMID- 8422008 TI - Lactate dehydrogenase in CML. PMID- 8422009 TI - Large cell calcifying Sertoli cell tumor of the testis. PMID- 8422010 TI - Expression of matrix metalloproteinase-2 related to lymph node metastasis of oral squamous cell carcinoma. A clinicopathologic study. AB - To investigate the effect of matrix-degrading enzymes on the malignant potential of oral squamous cell carcinoma, the expression of matrix metalloproteinase-2 (MMP-2/72-kD gelatinase/type IV collagenase) in 46 patients who had neck surgery for oral cancer was studied immunohistochemically. In 20 of 26 patients (76.9%) with lymph node metastases, proMMP-2 was strongly expressed, whereas the production of proMMP-2 in tissue was detected only in 5 of 20 patients (25%) who had no lymph node metastases. In tissue specimens, proMMP-2 was expressed in a diffuse invasive mode and in the advancing front of cancer. Because MMP-2 can degrade type IV collagen composed of basement membrane, these results suggest that the in vivo production of the enzyme by cancer is an indicator of the degree of malignancy, and that the analysis of proMMP-2 expression is useful to evaluate the malignant potential in individual oral squamous cell carcinoma. PMID- 8422011 TI - Focal enterocyte vacuolization. A new microscopic finding in the acquired immune deficiency wasting syndrome. AB - Vacuolization of duodenal enterocytes was found by light microscopic examination in five patients meeting the Centers for Disease Control criteria for the acquired immunodeficiency wasting syndrome. Four of these patients had chronic diarrhea and malabsorption as documented by an abnormal D-xylose test, whereas one patient had no diarrhea or malabsorption. Enterocyte vacuolization was patchy in distribution, although affected cells were most notable on villous tips. Staining with period acid-Schiff, acid-fast bacilli, periodic acid-Schiff following diastase treatment, Congo red, and alcian blue were negative, suggesting that vacuolization is due to lipid accumulation. Immunoperoxidase staining for the human immunodeficiency virus envelope protein gp41 was positive in lamina propria mononuclear cells in all five patients. The authors hypothesize that lipid accumulation represents an enterocyte response to injury, possibly by an indirect effect of the human immunodeficiency virus. PMID- 8422012 TI - Immunohistochemical identification of receptors for epidermal growth factor in tumor endothelium may be affected by cross-reactivity to blood group A antigen. AB - It has been reported that endothelium in malignant glioma stains with a commercial antibody raised against the receptor for epidermal growth factor (EGFr) on A431 cells (clone 29.1). In this report, this antibody was used to study the immunohistochemical expression of EGFr in benign and malignant ovarian, mid-gut carcinoid, and thyroid neoplasms using the avidin-biotin-peroxidase complex technique. Eighteen of the 37 ovarian neoplasms, 4 of the 10 thyroid neoplasms, and 14 of 28 mid-gut carcinoid tumors expressed strong and distinct endothelial staining, whereas staining results of the remaining tumors were negative. The endothelial nature of the staining was verified by staining serial sections with Ulex europaeus agglutinin-I. The staining was independent of that obtained with an antibody raised against a synthetic peptide consisting of residues 985 to 996 from the cytoplasmic domain of EGFr (clone F4). All positive staining occurred in patients determined to be of blood groups A or AB, whereas samples from patients with blood groups B or O were negative. Immunoabsorption of the antibody with centrifuged erythrocytes from a blood group A donor, but not from a blood group B donor, abolished the positive staining. The data indicate that positive staining of tumor endothelium with this antibody is due to cross reactivity with blood group A antigen. The results obtained challenge the validity of previously performed immunohistochemical studies in which monoclonal antibodies raised against the EGFr of A431 cells have been used, and in which the epitope for the monoclonal antibody has not been determined. PMID- 8422013 TI - Improvement of the quantification of estrogen and progesterone receptors in paraffin-embedded tumors by image analysis. AB - It has been shown previously that estrogen receptors (ER) detected by immunohistochemical examination of paraffin-embedded tissue sections could be quantified by computerized image analysis. Several factors were identified that were, in part, responsible for the modest correlation obtained with biochemical assay results. In the present study, 45 formalin-fixed, paraffin-embedded breast carcinomas from a previous study were reevaluated to determine if current methods could provide a better correlation and more consistent results. Sections of the tumors were made to react with estrogen and progesterone receptor antibodies (ER ICA and PgR-ICA) and the intensity of the stain was quantified using an image analysis system. Vimentin immunostain was used to assess the degree of antigenic loss. Quantitation was performed only on the areas with nuclear staining. The correlation of the estrogen and progesterone receptor values obtained by the dextran charcoal-coated method with the percentages of stained areas and with the intensity of the stain was excellent. The agreement between both methods was 91.1% for estrogen and 86.7% for progesterone receptor values. These results represent a significant improvement compared with those found in a previous study (87% agreement for estrogen receptor). The current approach to estrogen and progesterone receptor quantitation is simplified and eliminates subjectiveness in the selection of the fields for evaluation. The studies are reproducible because discrepancies due to sampling techniques are excluded. Finally, the method validates the technique as a substitute for cytosol-based methods. The results of the two vastly dissimilar assays, the pitfalls of the "gold standard" dextran charcaol-coated assay, and the need for retrospective studies to acquire a range of quantified ER-ICA and PgR-ICA values with clinical significance are discussed. PMID- 8422014 TI - Progressive hemochromatotic cardiomyopathy despite reversal of iron deposition after liver transplantation. AB - Dilated cardiomyopathy is a frequent and serious complication of idiopathic hemochromatosis. The mechanism by which disordered iron metabolism induces heart failure is not entirely understood, but myocardial dysfunction appears to be intimately related to the deposition of iron in myocytes. Cardiac function characteristically worsens or improves in proportion to the degree of iron accumulation in cardiac myocytes. The authors report the case of a 47-year-old man with idiopathic hemochromatosis and cirrhosis who developed symptoms of congestive heart failure and was found to have dilated cardiomyopathy 7 months after receiving a liver transplant. An initial endomyocardial heart biopsy demonstrated severe iron deposition in myocytes. The patient's heart failure worsened in the next 3 years and he eventually required a heart transplant. Examination of the explanted heart revealed dilated cardiomyopathy, but the previously demonstrated iron deposits in the cardiac myocytes were depleted. This "uncoupling" of cardiac function and cardiac iron load suggests that a threshold may be reached at which point the metabolic and ultrastructural derangements of iron deposition are no longer reversible, even with the removal of the inciting agent. Furthermore, displacement of myocyte iron stores after liver transplantation implicates altered hepatic iron metabolism as a primary or contributing mechanism in the pathophysiology of idiopathic hemochromatosis. PMID- 8422015 TI - Capturing spirochetes from humans. PMID- 8422016 TI - Cervicovaginal cytology, false-negative results, and standards of practice. PMID- 8422017 TI - Equipment, standardization, and applications of image processing. PMID- 8422018 TI - Changing etiology of macrocytosis. Zidovudine as a frequent causative factor. AB - Macrocytosis is most commonly associated with vitamin B12 and folate deficiencies, followed by alcoholism, liver disease, and malignant neoplasms. Many laboratories have observed that in recent years macrocytosis increasingly has been associated with zidovudine treatment of acquired immune deficiency syndrome. One hundred consecutive inpatients in a large metropolitan urban hospital with mean corpuscular volumes greater than 110 fL were studied; 44% were patients with acquired immune deficiency syndrome being treated with zidovudine, 19% were alcoholics, and 12% had malignant neoplasms. Only 3% were folate deficient and just 4% were vitamin B12 deficient. This study suggests that zidovudine has become the most common cause of macrocytosis in the hospitalized urban patient population and that vitamin B12 and folate deficiencies have decreased in proportion. PMID- 8422019 TI - Inosithin neutralization test to measure lupus anticoagulants. AB - The inosithin neutralization test was performed in 14 patients in whom lupus anticoagulant was detected. To test its specificity, it was also performed in 10 patients with severe hemophilia and in three patients with Factor VIII inhibitors. Prolonged kaolin clotting time was corrected by adding varying amounts of inosithin (Asolectin, 0.19 to 100 micrograms), a soybean-derived phospholipid, in all patients with lupus anticoagulant but not in patients with hemophilia or in two patients with Factor VIII inhibitors. In one patient, both Factor VIII inhibitors and lupus anticoagulant were present. The concentration of lupus anticoagulant in a patient was expressed as the amount of inosithin (measured in micrograms) required to normalize the prolonged kaolin clotting time. This amount correlated significantly with the occurrence of thrombosis and/or recurrent abortion. The inosithin neutralization test is useful to measure lupus anticoagulant. PMID- 8422020 TI - Mean platelet volume during coagulase-negative staphylococcal sepsis in neonates. AB - Coagulase-negative staphylococci are the most common cause of late-onset septicemia in neonates in intensive care nurseries. Clinical and laboratory diagnosis of infection with coagulase-negative staphylococci can be difficult. The authors reviewed serial mean platelet volumes of 18 infants in whom coagulase negative staphylococci sepsis developed and found a significant increase in the mean platelet volume at the time of diagnosis and a return to baseline after resolution of the infection. The increase in mean platelet volume occurred although thrombocytopenia developed in only two of the infants and no difference was found in the mean platelet counts before and at the time of diagnosis of the infection. This finding may be a useful adjunct to the current laboratory tests used to diagnose coagulase-negative staphylococci sepsis in neonates. PMID- 8422021 TI - Evaluation of the Coulter STKS five-part differential. AB - The authors evaluated the Coulter STKS (Coulter Corp., Hialeah, FL) five-part differential in a tertiary-care hospital using samples with a broad range of distributional and morphologic abnormalities. Particular attention was given to the performance of the instrument-generated suspect flags that occur as an aid to identify samples with abnormal leukocytes. A morphologically abnormal, or positive, blood smear was defined by the presence of any blasts, malignant lymphoid cells, grossly dysplastic neutrophils, nucleated red blood cells (nRBC), platelet clumps, or reactive lymphocytes of more than 5%. The presence of any white blood cell-related suspect flag, except for Immature Granulocyte/Bands (i.e., Blasts, Variant Lymph, NRBC, Platelet Clumps, Review Slide, or WBC*R), was considered to be a positive instrument result. The STKS showed excellent quantitative results for the WBC differential compared with the manual differential when these "morphologic abnormalities" were absent in a 400-cell manual differential or low in numbers (< or = 5%). Specificity of these non immature granulocyte/band suspect flags was good, with a false-positive rate of only 11.7%. Overall sensitivity in 113 samples with morphologic abnormalities was 67.3%. Sensitivity to detection of > or = 1% abnormal WBCs or > or = 1 nRBC/100 WBCs (a subset of 78 samples) was 80.8%. Sensitivity to detection of more than 5% abnormal WBCs or more than 5 nRBC/100 WBCs (a subset of 53 samples) was 84.9%. The primary deficiency was the inability of the STKS to flag samples with lymphoma cells, lymphoid blasts, or more than 5% reactive lymphocytes. PMID- 8422022 TI - An unusual case of mycosis fungoides presenting as sarcoidosis or granulomatous mycosis fungoides. AB - The authors describe a case of cutaneous and lymph node granulomas first reported as sarcoidosis. As skin sarcoidlike reactions disappeared, the development of typical histologic and immunopathologic features of cutaneous mycosis fungoides suggested granulomatous mycosis fungoides. This case illustrates the difficulties in differentiating true systemic sarcoidosis associated with mycosis fungoides from sarcoidlike reactions when extensive granulomas obscure the underlying cutaneous lymphoma. This report emphasizes the utility of immunohistochemical analysis to identify the early cutaneous T-lymphomatous infiltrate, initially admixed with epithelioid and giant cell granulomas. This technique also made it possible to characterize a Ki-1-positive anaplastic large-cell lymphoma when the transformation of mycosis fungoides into highly malignant lymphoma occurred in the lymph node. PMID- 8422023 TI - Sequential measurement of anti-platelet antibodies in a patient who developed EDTA-dependent pseudothrombocytopenia. AB - Ethylenediaminetetraacetic acid (EDTA)-dependent pseudothrombocytopenia is the occurrence of a falsely low platelet count caused by antibodies that agglutinate platelets in the presence of EDTA. If unrecognized, it may result in the erroneous diagnosis of thrombocytopenia and possible inappropriate therapy. It has been noted that this phenomenon tends to appear in hospitalized patients after an initially normal platelet count, but sequential measurements of anti platelet antibody have not been reported. The case of a patient who developed EDTA-dependent pseudothrombocytopenia approximately 1 week after being hospitalized for severe trauma is described. Anti-platelet antibodies were not detected on admission by a radiolabeled antiglobulin technique but were shown to increase in titer concurrent with the appearance of EDTA-dependent pseudothrombocytopenia. PMID- 8422024 TI - Identification of false-positive CK-MB activity in an elderly patient. AB - An unusual occurrence of an aberrant CK-MB activity in an elderly female patient is described. The CK-MB-like activity present in serum samples from this patient was definitively identified as an immunoglobulin A complex of CK-MM (macro CK, type 1) using CK isoenzyme agarose gel electrophoresis after pretreating the patient's serum sample with either anti-immunoglobulin A antiserum or anti-CK-M antibody reagent. PMID- 8422025 TI - A severe mandibular discrepancy case. PMID- 8422026 TI - Stability of mandibular constriction with a symphyseal osteotomy. AB - The purposes of this study are to determine the stability of surgical mandibular constriction with a midline osteotomy and to evaluate the periodontal and temporomandibular joint responses. A symphyseal osteotomy to facilitate mandibular construction was performed in 15 patients to correct transverse discrepancies. At the same time a surgical anterior or posterior repositioning of the mandible was done by using a bilateral sagittal osteotomy of the vertical ramus. Tomograms in the coronal plane including the mandibular second molars were taken preoperatively (T1), immediately postoperatively (T2) and 8 weeks postoperatively (T3). Linear measurements between the cortical borders of the mandible were assessed from the tomograms at each time period. Median surgical and postoperative changes in mandibular width were determined. When evaluating the entire group no statistically significant change in the surgical constriction was found postoperatively (T2 and T3), although there was some individual variability. An examination of the periodontal response at the osteotomy site revealed no statistically significant change between the initial and the 5-month postoperative examinations. No changes in joint noise were detected postoperatively, and all mandibular joint movements returned to preoperative values except for excursive movements. Mandibular constriction with a midline osteotomy on conjunction with a bilateral sagittal osteotomy was found to be a stable modality for correcting transverse disharmonies. PMID- 8422027 TI - Comparison of Roth appliance and standard edgewise appliance treatment results. AB - A retrospective comparison of Roth appliance and standard edgewise appliance treatment results was made using two indices. The first, the ideal tooth relationship index (ITRI), scored dental casts for the presence of ideal tooth contacts. The second judged posttreatment dental casts on the basis of criteria established by Andrews in his "Six Keys to Normal Occlusion." The sample consisted of 120 orthodontically treated cases completed by two practitioners who have used both the Roth and standard edgewise appliances. Thirty cases of each appliance type were collected from each practitioner. The overall posttreatment ITRI percentage scores showed no significant differences between the appliances. Practitioner differences existed for the anterior intraarch, anterior interarch, and posterior buccal interarch relationships. These differences were related to both treatment time and finishing arch wire size. The results of the Six Keys Analysis showed that the angulation and inclination of the maxillary posterior teeth were better with the Roth appliance. However, success in achieving some components of the six keys did not translate into an increased percentage of ideal tooth contacts as measured by the ITRI. Despite using the Roth appliance, experienced clinicians still found it difficult to achieve all six keys to normal occlusion. PMID- 8422028 TI - The use of twins in dentofacial genetic research. AB - A literature review is given on the different results obtained with twin and family studies in relationship to the development and structure of the dentofacial complex. Recent advances in twin and family studies are mentioned. Attention is focused on functional components considered to be of primary importance in craniofacial growth. PMID- 8422029 TI - Genioglossi muscle activity in response to changes in anterior/neutral head posture. AB - Clinicians have acknowledged swallowing, tongue activity, and head posture as interdependent variables that must be concurrently examined. The purpose of this study was to evaluate genioglossus activity during swallowing, rest, and maximal tongue protrusion in two head positions (HPs) with a noninvasive recording device. Eight Angle Class I subjects were evaluated. Repeated measures were performed in a single session to record surface intraoral electromyographic (EMG) activity of the genioglossus muscles. Head position was measured in angular degrees from photographs. Three variables were measured in both the neutral-head position (NHP) and anterior-head position (AHP): (1) duration of genioglossus EMG during swallowing, (2) genioglossus EMG with the tongue at rest, and (3) genioglossus EMG during maximal isometric tongue protrusion. A Wilcoxin matched pair signed-rank statistic was used for EMG analysis, and a paired sample t test statistic was used for head posture analysis. The angles measured for NHP and AHP within each subject were significantly different verifying two different head positions. Duration of swallowing was not significantly different between head positions. Resting genioglossus EMG and maximal isometric genioglossus EMG were statistically greater in the AHP. The data suggest that head positional changes may have an effect on genioglossus muscle activation thresholds. However, small differences in resting EMG activity between head positions suggests that the clinical significance needs further investigation. PMID- 8422030 TI - The effect of fixed and functional appliances on enamel decalcifications in early Class II treatment. AB - The presence of enamel decalcifications before and after early Class II treatment was examined on 41 subjects and 164 permanent first molars. No new demineralization areas were found in patients wearing only a removable appliance. Of the maxillary molars 6% displayed decalcifications in patients wearing a headgear only: whereas of patients wearing a headgear and biteplate 25% displayed enamel lesions. Eighteen percent of white spot lesions diagnosed before treatment had shown a "reversal phenomenon," whereby the enamel appeared normal after treatment. This process only occurred in the functional appliance group indicating that remineralization was better achieved with an appliance that was capable of being removed from the oral cavity. PMID- 8422031 TI - Maxillary incisor crown-root relationships in different angle malocclusions. AB - The long axis of the maxillary incisor root is not always identical to that of the crown. Instead, there is appreciable variation in the crown-root angle, generally with the crown torqued lingual to the root axis. In orthodontic cases assessed before and at the end of full-banded treatment, the crown-root angle was significantly deflected in the Class III molar relationship series, notably so in moderate to severe cases where the maxillary incisors are constrained lingual to the lower arcade. Apical root resorption was not significantly associated with the crown-root angle before or after comprehensive orthodontics. Cephalometric predictors of the amount of deflection of the crown-to-root axis were localized to intertooth relationships (overjet, interincisal angle). It is proposed that the large collum angles in Class III cases develop during tooth eruption when the maxillary incisors are trapped within the lower arch; this torques the crown of the maxillary incisor but leaves the unmineralized portion of the root free to develop as if the crown were still in its prior, more procumbent orientation. PMID- 8422032 TI - A study of clinical signs and symptoms of temporomandibular dysfunction in subjects with normal occlusion, untreated, and treated malocclusions. AB - In the light of the universally growing concern over the suggested relationship between orthodontic treatment and temporomandibular (TM) dysfunction, the purpose of the present investigation was to compare the status of signs and symptoms of TM disorders in three groups of adolescents and young adults. The groups consisted of 30 persons with normal occlusions, 41 with untreated malocclusions, and 31 with treated malocclusions. The clinical status and subjective symptoms of TM dysfunction were recorded according to the principles introduced by Helkimo. The results showed that the normal occlusion group had the maximum number of persons free from any dysfunction, but the differences between the groups in the distribution of persons according to the anamnestic and clinical dysfunction indices were not significant. The only statistically significant finding was the difference in the clinical dysfunction index scores of the persons with normal occlusions and untreated malocclusions. According to anamnesis, the most frequently reported symptoms were related to periods of stress. Among the clinical signs and symptoms, the most commonly occurring were crepitations on palpation and sounds on auscultation of the joints in all the three groups. In conclusion, the absence of substantial differences between the three groups indicates that the role of orthodontic treatment in either precipitation or prevention of TM dysfunction remains questionable. PMID- 8422033 TI - Root resorption after orthodontic treatment: Part 1. Literature review. PMID- 8422034 TI - Clinical excellence in an academic program? PMID- 8422035 TI - Orthodontic education. Portending its future? PMID- 8422036 TI - A simple removable acrylic-free retainer (the Sarhan type). PMID- 8422037 TI - Biodegradation of orthodontic appliances. Part I. Biodegradation of nickel and chromium in vitro. AB - The purpose of this study is to compare in vitro the corrosion rate of a standard orthodontic appliance consisting of bands, brackets and either stainless steel or nickel-titanium arch wires. The corrosion products analyzed were nickel and chromium. Evaluation was conducted with the appliances immersed for 4 weeks in a prepared artificial saliva medium at 37 degrees C. Ten identical sets were used, each simulating a complete orthodontic appliance used on a maxillary arch with a full complement of teeth. Five sets were ligated to stainless steel arch wires, and the other five sets were ligated to nickel-titanium arch wires. Nickel and chromium release was quantified with the use of a flameless atomic absorption spectrophotometry. The analysis of variance was used to determine if differences existed between the nickel and chromium release according to arch wire type, as well as with time (days 1, 7, 14, 21, and 28). The results indicate that (1) orthodontic appliances release measurable amounts of nickel and chromium when placed in an artificial saliva medium. (2) The nickel release reaches a maximum after approximately 1 week, then the rate of release diminishes with time. On the other hand, chromium release increases during the first 2 weeks and levels off during the subsequent 2 weeks. (3) The release rates of nickel or chromium from stainless steel and nickel-titanium arch wires are not significantly different. (4) For both arch wire types, the release for nickel averaged 37 times greater than that for chromium. How much of these corrosive products are actually absorbed by patients still needs to be determined. PMID- 8422038 TI - Orthodontists and the ADA. PMID- 8422039 TI - Channels and vehicles of communication: the asbestos awareness campaign. AB - In April 1978, the U.S. Department of Health, Education, and Welfare launched a multimedia campaign to inform doctors, workers, and others about the increased risks of asbestos exposure. Unlike most worker notification efforts, this one has no lists of workers or even of workplaces but faced the challenge of locating people who had worked in the shipbuilding industries more than 30 years earlier. Multiple mass media channels were used, but since most messages were distributed as public service announcements, gatekeepers were critical to the success of the campaign. Some campaign messages were aired, but mostly at hours other than prime time, and the coverage focused more on the controversial, fast-breaking events rather than on estimates of risk or on behaviors to reduce risk. The campaign was effective in increasing the number of people who believed they were at risk, but was less successful with older Americans than with manual laborers. PMID- 8422040 TI - Worker notification: lessons from the past. AB - The methodological choices being made in current worker notification programs are influenced in both obvious and subtle ways by certain ground-breaking notifications carried out during the 1970s and 1980s. A lore has built up concerning these cases. Their names are heavy with implied meaning. As worker notification gains in acceptance and frequency, some of those newly interested in the area will be unfamiliar with these early cases, their significance, and their lessons. To help provide a context for these proceedings, six landmark worker notifications are briefly described here. Different cases might have been chosen; more cases might have been added. However, these six give a flavor of the struggles, trials, triumphs, and insights that have marked worker notification's passage through recent history. PMID- 8422041 TI - Social aspects of high-risk notification among chromium-exposed workers. AB - This study, based on 169 telephone interviews, explores how a sample of chromium exposed workers responded to notification of their cohort's elevated risk of lung cancer. It is important to recognize that notified workers do not react as isolated individuals. Their responses are social, actively constructed through interaction with others, unfolding over time within a context of relationships and shared symbols that mediate the risk information. This report illustrates some of the ways that socially-based beliefs and interaction with the social environment can influence worker response, and suggests a more sociologically sophisticated concept of notification to fit the realities of workers' lives. PMID- 8422042 TI - High-risk notification of chromate and bichromate production workers. AB - A high-risk notification program is in progress for 4,862 former chromate and bichromate production workers. A previous mortality study of this cohort found a 29-fold increase in mortality for respiratory cancer. The increase was greater in black than white workers (80-fold vs. 15-fold). The cohort was compiled from Social Security Administration records. Notification was performed mainly using addresses on file with the Internal Revenue Service. At this time, 68% of the cohort has either been determined to have died or has been contacted. A description of the various steps of notification is included. This project has demonstrated that it is possible to perform high-risk notification in an economical manner even in the absence of employer personnel records or current addresses. The methodology used in this project is generalizable to other cohorts. PMID- 8422043 TI - Comprehensibility of material safety data sheets. AB - The Hazard Communications Standard, commonly known as the Worker Right to Know law, requires that a chemical information sheet, known as a material safety data sheet (MSDS), be produced for all hazardous chemicals. This study examines the comprehensibility of a sample of MSDSs to a group of about 100 unionized workers in manufacturing industries located in the state of Maryland. Workers were given several MSDSs and performed the equivalent of an open-book test answering questions regarding health and safety information that was provided on the MSDS. On average, the information on the MSDSs was found to be about one-third incomprehensible. This result raises concerns regarding policies and practices for hazard communication. There is a clear indication that MSDSs need to be improved if workers must rely on them for health and safety information. PMID- 8422044 TI - Asbestos screening and education programs for building and construction trades unions. AB - Worker notification involves informing current and past employees of their risk of disease. It also involves suggesting ways to reduce their risks. The asbestos screening and education programs designed for the building and construction trades unions were a national multisite effort that focused on improving the health of eligible union members and retirees at high risk of developing asbestos related disease. The asbestos screening and education programs were made available to "high-risk," asbestos-exposed local union members through the efforts of a number of international unions, including the International Union of Elevator Constructors and the Laborers' International Union of North America- both affiliates of the Building and Construction Trades Department, AFL-CIO. Consultation and program assistance in developing and implementing these programs were provided by the Occupational Health Foundation, a labor-sponsored, nonprofit organization with a multidisciplinary safety and health staff. Program components included identification of "high-risk" individuals, notification of risk, education, medical screening, legal referral, and various support services. Community-based physicians and/or physician-staffed mobile testing units provided services on a contractual basis according to a standardized medical protocol. Between 1988 and 1991, 2,136 union members and retirees from 89 local unions affiliated with the Elevator Constructors or the Laborers were screened in 59 regional programs. A general description and historical perspective are offered concerning program implementation, integration into existing union infrastructures. Emphasis is placed on the role of the unions in advancing members' interests when dealing with the health and socioeconomic implications of asbestos-related disease. PMID- 8422045 TI - Beyond notification: a case study from Cloquet, Minnesota. AB - All members of Local 158 of the United Paperworkers International Union in Cloquet, Minnesota, were invited to attend a meeting to notify them about the outcome of the Union's study concerning exposure to asbestos fiber. Union members and their families were contacted by letter, radio and television announcements, area newspapers, and word of mouth. They were informed about the medical screening that was being planned and about other information on asbestos-related diseases. During the meeting, they were asked to attend one of the educational sessions, to schedule an appointment for the screening, and to fill out a medical history. It was determined that paperworkers with asbestos-related disease might have rights to at least two types of compensation: worker's compensation from their employers and/or court- or jury-awarded compensation from asbestos manufacturers. Other sources of compensation such as social security of disability pensions might also be available to eligible individuals. The Union did not have a complete listing of all the individuals who worked at plants or in operations where asbestos exposure may have occurred. The Union solicited the aid of the Minnesota Department of Health to help identify former asbestos-exposed workers and to develop an ongoing screening and medical program for workers who were exposed but did not develop symptoms of asbestos-related disease; these illnesses usually take 20 years or more to develop. However, the Union was able to follow-up the notification and subsequent screening by sending members and their families the results of the initial screenings and other information concerning the status of any legal actions taken on behalf of the exposed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422046 TI - Experiences of a state-sponsored notification and screening program for asbestos workers. AB - Worker notification can involve a broad range of activities including medical screening, personal and mass communications, cohort identification and tracing, and even litigation. The inclusion or exclusion of various supporting activities in a worker notification program may pose significant medical, public health, financial, logistical, and even legal implications for targeted individuals as well as for the agencies involved. This report describes some experiences in a state-sponsored notification and screening program of approximately 4,500 asbestos workers in Minnesota. In this program, a variety of factors led to the decision to provide medical screening to 1,101 workers and 451 spouses. It is anticipated that another 3,400 workers will be notified but not screened. A follow-up survey of notified workers showed overwhelming support for this program. It is estimated that this program will cost more than $650,000 by its completion. The decision to institute medical screening and other support activities should be made with careful consideration of the diverse implications of these activities to the individuals, communities, and agencies involved. PMID- 8422047 TI - Notification in the community context. PMID- 8422048 TI - Dioxin: a case study. AB - The need to notify individuals of a possible health risk from their past exposure to potentially hazardous agents frequently extends beyond workers to include community groups. The issues to consider in community notification are frequently similar to those that are important for worker notification but may include some that are unique. This case study traces the evolution of one company's strategy for communicating with the public about possible dioxin contamination associated with its operations. Early communications tended to emphasize the technical aspects of the issues in the fashion of scientists talking to other scientists. This was interpreted by some to be symptomatic of an arrogant and uncaring attitude. Beginning in the early 1980s, the company's management recognized the need to reach out to a variety of audiences on multiple levels, and shifted to a more comprehensive communications strategy. A similar shift is now occurring throughout the chemical manufacturing industry as top managers realize that, if they expect to continue to operate, they must become more accountable and responsive to the public. PMID- 8422049 TI - Superfund and one community program. AB - The U.S. Environmental Protection Agency--Region II conducted a pilot program in risk communication and notification of hazardous waste information at a Superfund site in Toms River, New Jersey. The program was designed to assess the levels of awareness and concern among local citizens and to provide accurate information about health risks associated with potential exposure to environmental contaminants. The purpose of this program was to develop linkages among the community; local, state, and federal governments; industry; health professionals; and educators in dealing with environmental problems. A Community Leaders Network was formed and became actively involved in the program. Following a community needs assessment for risk information, a series of fact sheets was developed, pretested, disseminated, and evaluated. The analysis of the program highlights the important lesson of being able to respond to the specific changing dynamics of the community and offers guidelines useful for risk communication in many communities. PMID- 8422050 TI - Community lead exposure. AB - In August, 1990, a regional transit authority began removing paint on two overhead bridges on a commuter railroad line running through a residential neighborhood of Lynn, Massachusetts, an industrial city north of Boston. The contracted work involved the sandblasting of existing bridge paint to prepare the surface for repainting. The sandblasting was conducted without shrouding or adequate worker protection. The bridge paint was not tested for lead content, nor were neighborhood residents informed of the operations prior to the commencement of the work. Following complaints by residents to the local and state health departments, a cease-work order was issued. Laboratory tests of paint samples revealed a lead content ranging from 5% to 15%. Sandblasting debris was evident throughout the adjacent neighborhood. What followed was a 3-month effort by state and local government officials and the transit authority and its contractors to assess, communicate, and remedy the damage done by the operations. Soil, dust, and blood samples were collected to assess environmental contamination and health effects. Notices were sent, meetings held, and informational sites established to inform community residents of the impact of the operations and remediation efforts. The local and Boston media provided extensive coverage of the incident and its aftermath. Remediation efforts involved the relocation of occupants from residences in proximity of the site and extensive clean-up of the interiors of affected homes by licensed lead abatement contractors. The cost of the cleanup was well over $1 million.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422051 TI - Evaluation issues in the Drake Chemical Workers Notification and Health Registry Study. AB - The Drake Chemical Workers' Health Registry combined notification of workers about bladder cancer risk with access to a free program for screening and diagnosis. Evaluation of the project has given rise to several findings and new research questions. Findings in this article illustrate the following evaluation issues: 1) studying the combination of strategies that are most effective and cost effective to notify workers of their disease risks, 2) determining the realistic yield from strategies to gain participation in health screening and other protective services for notified workers, 3) identifying the notification strategies that were most effective for different kinds of participants, 4) using process evaluation to identify key activities for ensuring continued participation of cohort members in screening, and 5) examining the extent to which participants are willing to quit smoking to protect their health. PMID- 8422052 TI - Importance of evaluating the context within which notification occurs. AB - Workers' response to notification about health effects from exposure to toxic materials is determined not just by the content of the message but also by the "context" within which notification occurs, that is, the workers' prenotification knowledge, attitudes, and experiences concerning environmental health risks in general as well as the health problem that is the subject of the notification. In many cases, workers already have a high level of awareness of environmental health issues and are also aware that their work environments could be bad for their health, before receiving official notification of a particular health risk. This is one reason why worker notification programs often have limited impacts on the workers' health behaviors. To understand this process, researchers should assess workers' baseline attitudes and behaviors and study how they affect response to notification. Persons conducting notification programs should evaluate workers' prenotification attitudes, knowledge, and behaviors, and use this information in planning notification efforts. PMID- 8422053 TI - Evaluating the clarity of research reports written for research subjects. AB - It is not simple to report research results to the people who participated in the research as subjects of study. Few evaluations have examined subjects' understanding of written research findings. In two recent studies of solvent exposure, subjects received brief summaries of the research. Before they were sent, the summaries were evaluated by research, health, and communication specialists as well as by representatives of the target audience. For one of the studies, interviews were conducted with subjects before and after they received the reports. Although the subjects said they understood the summary, their answers to questions about its content indicated otherwise. The results suggest that researchers not rely on subjects' self-assessments of their understanding but instead design methods to test subjects' knowledge directly. Those tests would be most valuable if applied in formative evaluations, when the opportunity exists to improve the research summaries. PMID- 8422054 TI - Assessing quality of life when planning and evaluating worker notification programs: two case examples. AB - Notifying workers that they have been placed at risk for serious medical problems may affect their lives in unexpected ways. Although very little evidence supports the fear of notification causing grave psychological problems, more subtle disruptions in quality of life may result. In addition, psychological reactions to notification, such as anxiety or denial, may impede or enhance workers' abilities to process, remember, or act upon the information presented. Measuring how different notification strategies affect quality of life is crucial; this process can enable us to design and select effective strategies that improve, rather than disrupt, the lives of workers and their families. Two preliminary efforts to include consideration of quality of life in planning and evaluating notification programs are described. PMID- 8422055 TI - Is the medical community ready for worker notification? AB - Although the potential for early detection of disease has been of central importance in the evolution of worker notification policy and practice, it is not the only positive outcome of medical intervention. The provider-patient encounter also creates an important opportunity for patient-centered risk communication that may result in workplace modifications as well as lifestyle changes to reduce the likelihood of subsequent illness and injury--both work and nonwork related. At the same time, it may signal the beginning of a series of social, legal, and political actions that may have positive or negative consequences for notified workers. This article identifies several important roles for members of the medical community relating to worker and community notification programs and explores the readiness of health care professionals in assuming these roles. It is suggested that health care providers lack both the training and sociopolitical sensitivity needed for meaningful participation in the notification process. This article addresses possible short-term and long-term approaches to enhancing the readiness of health care providers and suggests that effective intervention with notified workers requires a willingness to step beyond the confines of the medical model. PMID- 8422056 TI - Methodologic issues in risk communications to workers. AB - Until the late 1980s, epidemiologists in general did not individually notify subjects of the results of epidemiological studies. Now that they are beginning to do so, the question arises of how best to notify those involved. In general, the methods, the processes and the policies related to effectively communicating risks to workers have not been thoroughly examined in the scientific literature. This is especially true in situations where workers have already experienced the exposures that led to increased risks for disease. The recent increasing numbers of notifications have raised several methodologic issues, which are examined in terms of: (1) the content of notification, (2) the process of notification, and (3) the evaluation of the impact and effectiveness of notification. Too often in the discussion concerning notification, attention is paid to the content but the process and evaluation are rarely considered. The potential impact and effectiveness of notification have been raised as reasons for or against notification, but rarely has there been a concerted effort to evaluate a notification in this regard. This workshop was designed to address all these issues. The ultimate goal is to improve communications for workers. PMID- 8422057 TI - Worker notification: viewpoint of the employer. AB - Prior to notifying workers of risk exposure, employers should first consider and weigh the psychological, physiological, and social costs as well as the potential negative impact on labor-management relations, community resources, and the costs of false negatives and false positives. Only if the contributions of notification override and outweigh the composite costs should employers feel compelled to notify workers. PMID- 8422058 TI - Cultural and linguistic factors in worker notification to blue collar and no collar African-Americans. AB - This presentation offers historical and sociolinguistic perspectives regarding the origins of the language used by African-Americans. U.S. descendants of slaves of West and Niger Congo African origin should not be thought of as native speakers of English. To the extent that they have lived in de jure or de facto social isolation from Euro-American English-speaking persons, they have retained a West and Niger Congo African thought process or substratum in the phonology and morphosyntax of their speech. Their mother tongue has been described by several African-American linguists, psychologists, and ethnologists as Ebonics. In a sense, Ebonics refers to the paralinguistic verbal and non-verbal sounds, cues, and gestures that are systematically used in the process of communication by many African-Americans. As one consequence of historical social isolation, African Americans invariably score poorly on nationally standardized scales of "Standard American" English competence. Therefore, worker notification intended for African Americans, especially among the blue- and no-collar levels, that does not contain an Afro-centric perspective will likely not prove successful. The linguistic factors, which can severely impede or tremendously enhance the effectiveness of worker notification to these groups, are therefore vital and should not be ignored. Worker notification targeted for limited English-speaking or unlettered populations should include the development of materials and presentations in the language or primary medium best understood by the audience. Based upon research findings on the psycholinguistics of learning modal preference, the development of notification materials that target a learning channel that is audio oral or aural-oral is promoted. PMID- 8422059 TI - The ethics of choice in the struggle against industrial disease. AB - This article argues that broad recognition of the ethical basis for society's decisions about the reduction of the risks of workers, including notification, has been, is, and will be a more persuasive method of establishing programs implementing the rights to know and act in controlling the industrial vector in human disease than sheer power politics. If the argument is correct, then attention needs to be focused on the nature of the ethics of choice. The moral dialectic in the history of science and medicine is traced to the metaphysically assumed values of open and closed models of knowledge and action that determine how we weigh genetic and environmental factors in the process of choosing who shall live and who shall die of disease, with strong industrially generated vectors. The author suggests that we escape from the concepts of the closed systems of the past, whose grammar reflects values repugnant to the empowerment of those notified of risk, to open systems that enhance community-ecologic values of life and freedom. PMID- 8422060 TI - Issues in notification: reflections of a public health worker. AB - The objectives of notification are to inform individuals of research results so that they can make informed choices regarding health care and risk reduction behaviors, and to inform workers and employers of results and recommendations in order to take actions to improve workplace health and safety. Many questions confront researchers or public health workers engaged in notification efforts. Who should be notified? What information should be included in notification materials? When should notification occur? How and where should notification take place? What is the socioeconomic context in which notification is occurring? This article presents a public health worker's perspective on these issues. PMID- 8422061 TI - Reading, readability, and legibility research: implications for notification letters. AB - This article defines communication science and then briefly describes three research areas relevant to worker notifications: (1) receiving of notification messages, (2) reading and understanding of notification messages, and (3) influences or effects of notification messages on workers. Next, the article focuses on the reading and legibility research relevant to worker notifications and then provides 16 guidelines for drafting worker notifications. Finally, the article suggests evaluating draft notifications by using one or more of the following: skilled editorial reviews, readability scoring, Cloze techniques, signal stopping techniques, in-depth personal interviews, focus groups, and usability testing. PMID- 8422062 TI - Content of worker notification. AB - This article summarizes discussions held during the workshop on the issue of the content of worker notification messages and related topics. The workshop itself involved a number of sessions where participants formed small groups. This article represents a distillation of the summary reports by the rapporteurs from each group. It is not intended as a complete review of this issue. Rather, it presents some of the concerns that arose during freewheeling discussions, with the idea that the points raised may help others to think about some of the issues involved. PMID- 8422063 TI - Perspective on the content of worker notifications. AB - The concept of worker notification has evolved over the years to a prevailing view that workers should be notified of all epidemiologic studies that make use of their medical information. To adequately communicate health information to workers, epidemiologists must consider the intended audience along with its need for risk information that is scientifically valid and clearly outlines areas of uncertainty. This goal is facilitated by full disclosure to workers of information pertinent to the planning, conduct, and interpretation of research efforts. Subjectivity in choosing unanticipated research findings for inclusion in worker notifications may permit bias toward a personal viewpoint, but this can be minimized by prestudy agreement on appropriate criteria for selecting which research findings to communicate. Epidemiologic theory and principles of causal inference should guide the development of appropriate criteria. The timeliness of worker communications has received limited attention, and workers have often been the "last to know" about important studies. This may influence workers' receptivity to the risk message. Sponsoring organizations should ensure that an acceptable communications plan is included in research protocols and that the plan accords priority to notifying workers about study results. PMID- 8422064 TI - Notification of workers about an excess of malignant melanoma: a case study. AB - In January 1991, NIOSH completed a retrospective cohort mortality study of 3,588 Westinghouse Electric Corporation workers who had been engaged in the manufacture of electrical capacitors. The study evolved from a NIOSH Health Hazard Evaluation, which was conducted at the request of the Indiana State Board of Health because of its concern about PCB exposures among the Westinghouse workers. Life table analysis revealed a fourfold excess of deaths due to malignant melanoma. Though the workers were principally exposed to PCBs, the available exposure data did not lend itself to constructing an exposure-response curve that could relate PCB exposure to development of malignant melanomas. This was further complicated by the lack of substantial corroboration from other studies of PCB exposed cohorts. Because of the magnitude of the malignant melanoma excess and the fact that malignant melanoma is probably more amenable to treatment and remediation than most other cancers, NIOSH determined that notification of the individual cohort members was a prudent and necessary public health action. This article describes the notification process from the time the decision to notify was made through the postnotification period. It details the interaction between NIOSH, the former and current plant owners, the two labor organizations that represented the workers at the plant, and the recipients of the notification materials. Scientific and other issues surrounding this notification effort are also discussed. A number of lessons were learned about the notification process; these are described for the benefit of others who conduct notifications. PMID- 8422065 TI - Content of notification: exposure to the pesticide chlordimeform. AB - A notification about exposure requires a detailed search of records and interviews with personnel to properly identify potentially exposed groups. Next, these potentially exposed groups must be examined to determine which groups had the potential for highest exposures. The notification should address three key questions: Why am I receiving this notification? What problems are associated with exposure to the chemical? What am I supposed to do? Consideration should be given to notifying lesser-exposed groups, as to why they are not considered to be at increased risk. PMID- 8422066 TI - Asbestos disease risk communication conducted by the New Jersey Department of Health. AB - Since October 1985, the New Jersey Department of Health (NJDOH) has required hospitals to report all patients with a discharge diagnosis of asbestosis. As follow-up to a needs assessment survey of these patients, the NJDOH developed an educational packet including an information bulletin titled "Asbestos Disease: Medical and Legal Facts for Employees" and a pre-stamped postcard evaluation survey. The packet was sent to 1,418 patients reported by hospitals; 433 patients returned the evaluation questionnaire. The survey assessed attitudinal responses (were the materials helpful and easy to understand?) and behavioral responses (did the respondent plan on discussing the materials with a doctor or lawyer?) to the information in the packet. Of the postcard respondents, 85% found the materials helpful, and approximately half indicated that they would discuss the materials with a doctor or lawyer. Of the 33% who were proxy respondents, half appeared not to have understood that the materials had relevance to them as family members. Modifications to the protocol for the asbestos disease educational packet were made based on the evaluation data. Limitations of the evaluation survey and the value of process evaluations in risk communication projects are discussed. PMID- 8422067 TI - Practice guidelines. PMID- 8422068 TI - Cognitive remediation in schizophrenia: is it time yet? AB - Considerable effort has been directed toward identifying and conceptualizing information-processing deficits in schizophrenia and other psychotic disorders. The impressive gains in this field have made meaningful contributions to our understanding of the etiology and course of these disorders. This article considers whether the time is ripe to move beyond identification of these deficits to remediation of them. This move is far from a simple matter; it requires careful attention to theoretical frameworks and to the criteria for selecting certain cognitive deficits, among many, for remediation. A sparse and somewhat dated literature suggests that certain types of cognitive remediation, at least when defined in a narrow sense, are feasible with schizophrenic patients. This literature can reasonably justify continuing investigations on a modest scale. Specific questions for further studies include the following. 1) Which cognitive models provide a framework to guide cognitive interventions? 2) Which specific functions or levels of information processing should be targeted by cognitive interventions? 3) Are cognitive interventions effective? 4) Does the remediation of basic cognitive deficits generalize? A major strength of this area of investigation is the testability of the questions. Several research designs are tentatively suggested. PMID- 8422069 TI - The concept of boundaries in clinical practice: theoretical and risk-management dimensions. AB - The authors systematically examine the concept of boundaries and boundary violations in clinical practice, particularly as they relate to recent sexual misconduct litigation. They selectively review the literature on the subject and identify critical areas that require explication in terms of harmful versus nonharmful boundary issues short of sexual misconduct. These areas include role; time; place and space; money; gifts, services, and related matters; clothing; language; self-disclosure and related matters; and physical contact. While broad guidelines are helpful, the specific impact of a particular boundary crossing can only be assessed by careful attention to the clinical context. Heightened awareness of the concepts of boundaries, boundary crossings, and boundary violations will both improve patient care and contribute to effective risk management. PMID- 8422070 TI - Diagnostic decision making in psychiatry. AB - The purpose of this article is to examine the consequences of and possible responses to uncertainty in psychiatric diagnosis. Uncertainty is inevitable because of the overlap in characteristics, or test results, between populations with and without a psychiatric disorder. As a result, there is never one correct method of identifying cases and noncases (i.e., case definition). In this paper principles of decision analysis and clinical epidemiology are used to develop a framework for thinking about the consequences of different diagnostic schema and for choosing among them. The framework illustrated here involves choosing an external validator, choosing a separator, and choosing a cutoff. This framework is applied to the problems facing three hypothetical researchers, and the consequences for their research of different diagnostic choices are explored. It is demonstrated how the relevance of research to clinicians and policy makers rests on the choice of the case definition process. The prevalent use of structured psychiatric interviews has not been accompanied by adequate attention to the problem of determining a diagnosis once the information is obtained. It is argued that more attention must be given to this process if we are to make optimal use of available resources for research and treatment. PMID- 8422071 TI - Practice guideline for eating disorders. American Psychiatric Association. PMID- 8422072 TI - Use of posttraumatic stress disorder to support an insanity defense. AB - OBJECTIVE: The authors examine the allegation that the diagnosis of posttraumatic stress disorder (PTSD) is frequently abused in the legal system as the basis for a defense of not guilty by reason of insanity. METHOD: Data for the investigation were drawn from a study of insanity pleas gathered from court records in 49 counties in eight states. Data on the 28 insanity plea defendants for whom PTSD was diagnosed before or immediately after trial were compared with data on 8,135 defendants whose insanity pleas were based on other diagnoses. RESULTS: Insanity pleas by defendants with diagnoses of PTSD constituted only 0.3% of the cases. There were few significant differences between the two groups on demographic variables, psychiatric histories, previous involvement in crime, or current charges. The defendants with PTSD were more likely to have been married, less likely to have been arrested as juveniles, and less likely to have been detained after trial. CONCLUSIONS: Contrary to previously expressed concerns, PTSD was infrequently associated with an insanity defense in the cases in this study. In the cases in which pleas based on PTSD were used, they were no more likely to succeed than pleas based on any other diagnosis. Defendants with PTSD-related insanity defenses differed little from other insanity defendants, contradicting the stereotype of the person who is driven by PTSD to commit crimes. The data do not support fears of widespread misuse of the diagnosis of PTSD in connection with the insanity defense. PMID- 8422073 TI - Childhood physical abuse and combat-related posttraumatic stress disorder in Vietnam veterans. AB - OBJECTIVE: Early trauma in the form of childhood physical or sexual abuse has been associated with adult psychopathology. The purpose of this study was to compare rates of childhood abuse in Vietnam veterans with and without combat related posttraumatic stress disorder (PTSD). METHOD: Premilitary stressful and traumatic events including childhood abuse and other potential predisposing factors were assessed in Vietnam combat veterans who sought treatment for PTSD (N = 38) and Vietnam combat veterans without PTSD who sought treatment for medical disorders (N = 28). Stressful and traumatic events including childhood physical abuse were assessed with the Checklist of Stressful and Traumatic Events and a clinician-administered interview for the assessment of childhood abuse. Level of combat exposure was measured with the Combat Exposure Scale. RESULTS: Vietnam veterans with PTSD had higher rates of childhood physical abuse than Vietnam veterans without PTSD (26% versus 7%). The association between childhood abuse and PTSD persisted after controlling for the difference in level of combat exposure between the two groups. Patients with PTSD also had a significantly higher rate of total traumatic events before joining the military than patients without PTSD (mean = 4.6, SD = 4.5, versus mean = 2.8, SD = 2.9). CONCLUSIONS: These findings suggest that patients seeking treatment for combat-related PTSD have higher rates of childhood physical abuse than combat veterans without PTSD. Childhood physical abuse may be an antecedent to the development of combat related PTSD in Vietnam combat veterans. PMID- 8422074 TI - Psychopathology and psychiatric diagnoses of World War II Pacific theater prisoner of war survivors and combat veterans. AB - OBJECTIVE: This study assessed current and long-term psychological and psychiatric sequelae of war trauma in World War II Pacific theater combat veterans, some of whom had been Japanese prisoners of war (POWs). METHOD: A group of 36 POW survivors and a group of 29 combat veterans, all of whom had seen fierce fighting and heavy unit casualties, were compared approximately 40 years later on psychological instruments assessing psychopathology constructs, negative mood states, and symptoms of posttraumatic stress disorder (PTSD) and on the computer-administered National Institute of Mental Health Diagnostic Interview Schedule. RESULTS: Although similar in personal backgrounds and in having suffered catastrophic war trauma, the two groups differed in the severity and type of psychiatric symptoms and in the occurrence of psychiatric disorders. Anxiety and depressive disorders were common in both groups, but there were differences in the frequency of PTSD diagnoses. Among the POW survivors, 70% fulfilled the criteria for a current diagnosis and 78% for a lifetime diagnosis of PTSD, compared to 18% and 29%, respectively, of the combat veterans. CONCLUSIONS: The findings point to the persistent nature of symptoms thought to be residuals of extraordinary stress and the relation between severity of psychiatric sequelae and characteristics of the stressors. PMID- 8422075 TI - Lifetime prevalence of panic states. AB - OBJECTIVE: Little is known about the prevalence of panic symptoms that do not meet criteria for panic disorder. This study was conducted to determine the prevalence of panic disorder, panic attacks, and limited-symptom attacks in the general population. METHOD: The authors identified a community-based sample of 1,683 randomly selected adults in 18 census tracts in San Antonio, Tex.; 1,306 of these subjects agreed to be interviewed with the Structured Clinical Interview for DSM-III. Subjects were classified as having panic disorder if they met DSM III-R criteria, as having panic attacks if they had attacks of four or more panic symptoms but did not have panic disorder, and as having limited-symptom attacks if they had attacks of fewer than four symptoms but no full-blown panic attacks. RESULTS: The crude lifetime prevalence rates were 3.8% for panic disorder, 5.6% for panic attacks, and 2.2% for limited symptom attacks. Women had higher rates of panic disorder and panic attacks than men, but the difference between men and women was not statistically significant for limited-symptom attacks. No statistically significant differences in rates between Hispanic and either non Hispanic white or black subjects were found. Non-Hispanic white subjects had higher rates of limited-symptom attacks than black subjects. CONCLUSIONS: The prevalence of limited-symptom attacks in this community-based study was 2.2%; black subjects had lower rates than non-Hispanic white subjects. Panic attacks appear to be at least as common as DSM-III-R panic disorder and, like panic disorder, are more common among women. PMID- 8422076 TI - Bereavement after homicide: a synergism of trauma and loss. AB - OBJECTIVE: The purpose of this study was to examine the intensity and relationship of trauma and responses to bereavement in family members after homicide. METHOD: The authors established an outpatient clinic that offered evaluation and supportive psychotherapy after the homicide of a family member. A standardized evaluation protocol was followed with 18 adults in order to detail variables of previous trauma, bereavement, and psychiatric disorder. Standardized measures of bereavement (Texas Revised Inventory of Grief) and trauma (Impact of Event Scale and Dissociative Experiences Scale) were also administered. RESULTS: As a group, the 18 adults were characterized by a high frequency of antecedent psychiatric disorder (N = 12), the homicide of a child (N = 12), and an intensely idealized attachment to the decreased, whose image of violent dying recurred as a disorganizing flashback and dream. The measures of bereavement and trauma showed generally higher levels of intensity in the 18 subjects in the present study than in normal subjects and other cohorts of bereaved subjects. CONCLUSIONS: For those who have lost a family member through homicide, recognition of a relatively specific pattern of dysfunctional responses of grief and trauma would promote early identification and psychiatric referral. PMID- 8422077 TI - Sexually assaultive male juveniles: a follow-up. AB - OBJECTIVE: The purpose of this study was to investigate the adult outcome of a group of male juveniles who committed sexual assault. METHOD: Nineteen sexually assaultive male juveniles and a comparison group of 58 violent juveniles were studied over an 8-year period through use of criminal records and clinical interviews. RESULTS: Although in adolescence the two groups were similarly violent, on follow-up those who had committed sexual assault were significantly more likely to commit adult sexual offenses. They also committed significantly more violent nonsexual offenses. Childhood sexual abuse, especially by females, was associated with adult sexual offenses. CONCLUSIONS: Sexually assaultive delinquents are at particularly high risk for subsequent violence. Hence, special efforts must be made to treat these delinquents in adolescence. Prevention of violent sexual behavior must include improved methods of detecting sexual abuse, especially that perpetrated by older females. PMID- 8422078 TI - Partial hospitalization (day treatment) for psychiatrically ill elderly patients. AB - OBJECTIVE: The purpose of this paper is to present initial findings from a retrospective chart review of geriatric day treatment patients in order to focus attention on this potentially important area, add to the limited database in this area, and generate hypotheses for future investigations. METHOD: Data were abstracted from the charts of 100 geriatric day treatment patients over a period of approximately 5 years (1985-1989). Descriptive, univariate, and multiple regression techniques were used to describe the patients and identify variables associated with their outcomes. RESULTS: The typical patient in this program was a widowed white woman in her 70s who suffered from a depressive disorder. During the initial treatment period (usually approximately 3 months), 57% of the patients experienced some clinical improvement. Variables associated with a favorable outcome included diagnosis of a mood disorder rather than a psychotic disorder, better initial functional status, greater initial social support, fewer stressful events during treatment, and longer duration of treatment. CONCLUSIONS: Geriatric day treatment can be effective and merits further study as a mode of treatment for psychiatrically ill elderly patients. PMID- 8422079 TI - Familial aggregation of female sexual orientation. AB - OBJECTIVE: The purpose of this study was to determine whether female homosexuality is familial and whether it is cofamilial with male homosexuality. METHOD: Subjects included 84 homosexual and 79 heterosexual female probands recruited through newspaper advertisements. Probands were asked about their siblings' sexual orientations and were asked for permission to contact siblings to confirm their reports. RESULTS: The authors were able to contact 60% of eligible siblings, and the information they provided about their sexual orientations confirmed that probands' reports were highly accurate. Homosexual probands had a significantly higher proportion of homosexual sisters according to four criteria for rating siblings' sexual orientations. Homosexual probands also had a higher proportion of homosexual brothers; however, this difference was not significant. CONCLUSIONS: Female homosexuality appears to be familial. Further research is required to resolve the question of whether female and male homosexuality are cofamilial. PMID- 8422080 TI - Pathological laughing and crying following stroke: validation of a measurement scale and a double-blind treatment study. AB - OBJECTIVE: This study was undertaken to test the reliability and validity of the Pathological Laughter and Crying Scale and the effectiveness of nortriptyline treatment for patients with emotional lability following stroke. METHOD: Eighty two patients with ischemic brain injury-54 who had been hospitalized with acute stroke and 28 others who requested treatment for pathological laughing and crying -were given standardized psychiatric and neurological assessments and then administered the Pathological Laughter and Crying Scale. The 54 acute stroke patients were used to evaluate the Pathological Laughter and Crying Scale, and the 28 patients with pathological emotional display were randomly assigned to nortriptyline treatment or placebo in a 6-week double-blind trial to assess the efficacy of a tricyclic antidepressant in treatment of this disorder. RESULTS: The interrater reliability on the Pathological Laughter and Crying Scale for a subgroup of 15 patients was 0.93, and the test-retest reliability of the scale was excellent. After 4 and 6 weeks of treatment, scores on the Pathological Laughter and Crying Scale showed significantly greater improvement in the 14 patients given nortriptyline than in the 14 given placebo. Although almost one half of these patients also had major depression, the improvement in emotional lability was independent of depression status. In addition, response to treatment was not significantly affected by lesion location or time since stroke. CONCLUSIONS: The severity of symptoms in pathological emotional display can be reliably quantified with the Pathological Laughter and Crying Scale, and treatment with nortriptyline can effectively ameliorate this emotional disorder. PMID- 8422081 TI - Depressive symptoms and the self-reported use of alcohol, caffeine, and carbohydrates in normal volunteers and four groups of psychiatric outpatients. AB - OBJECTIVE: The authors examined the relationship between depressive symptoms and the self-reported use of alcohol, carbohydrates, and caffeine in normal volunteers and four groups of psychiatric outpatients. METHOD: Outpatients and normal volunteers were given a questionnaire asking about their use of each of the three substances in response to each of the 14 depressive symptoms on the Hamilton Rating Scale for Depression. They also rated how much each substance improved each symptom. Twenty-six normal volunteers, 35 patients with major depression, 117 patients with seasonal affective disorder, 16 patients with alcohol dependence, and 24 patients with comorbid primary depression and secondary alcohol dependence completed the questionnaire. Test-retest reliability was established. Analysis of variance and stepwise multivariate discriminant function analyses were used to determine if diagnostic groups differed in the reported use and effect of each of the three substances. RESULTS: The responses concerning use and effect of alcohol of patients with alcohol dependence with or without depression were indistinguishable from each other. The responses of the patient groups regarding caffeine and carbohydrate use did not differ from each other, but all differed significantly from the responses of normal volunteers. Discriminant function analysis distinguished alcoholics from nonalcoholics in the relationship between drinking and the symptoms of anger and anhedonia. CONCLUSIONS: The relationship between symptoms and substance use varied depending on the substance. Alcoholics without depression were as likely to report drinking in response to depressive symptoms as were those who had had depression. Patients of all diagnostic groups were more likely than normal volunteers to report using caffeine and carbohydrates in response to depressive symptoms. PMID- 8422082 TI - Body dysmorphic disorder: 30 cases of imagined ugliness. AB - OBJECTIVE: Body dysmorphic disorder, preoccupation with an imagined defect in appearance, is included in DSM-III-R but has received little empirical study. The authors investigated the demographics, phenomenology, course, associated psychopathology, family history, and response to treatment in a series of 30 patients with the disorder. METHOD: The patients (including 12 whose preoccupation was of probable delusional intensity) were assessed with a semistructured interview and the Structural Clinical Interview for DSM-III-R, and their family histories were obtained. RESULTS: The 17 men and 13 women reported a lifetime average of four bodily preoccupations, most commonly "defects" of the hair, nose, and skin. The average age at onset of body dysmorphic disorder was 15 years, and the average duration was 18 years. Seventy-three percent of the patients reported associated ideas or delusions of reference; 73%, excessive mirror checking; and 63%, attempts to camouflage their "deformities." As a result of their symptoms, 97% avoided usual social and occupational activities, 30% had been housebound, and 17% had made suicide attempts. Ninety-three percent of the patients had an associated lifetime diagnosis of a major mood disorder; 33%, a psychotic disorder; and 73%, an anxiety disorder. The patients generally responded poorly to surgical, dermatologic, and dental treatments and to adequate trials of most psychotropic medications, with the exception of fluoxetine and clomipramine (to which more than half had a complete or partial response). CONCLUSIONS: This often secret, chronic disorder can cause considerable distress and impairment, may be related to obsessive-compulsive disorder or mood disorder, and may respond to serotonin reuptake-blocking antidepressants. PMID- 8422084 TI - Quality assessment and improvement in group psychotherapy. AB - OBJECTIVE: The authors sought a practical means of monitoring and evaluating group psychotherapy, using existing clinical resources, for purposes of quality improvement and education on a large general hospital psychiatric service. METHOD: Monitoring indicators were developed which addressed 1) the integration of group psychotherapy into treatment planning and 2) the competence and technique of group psychotherapists. The second indicator was assessed by skilled observers using a newly constructed Group Psychotherapy Rating Scale in direct observation of group psychotherapy sessions. The rating scale was examined for interrater reliability and, as a measure of construct validity, for its ability to distinguish the performance of professional staff therapists from that of trainees. RESULTS: The indicators provided useful monitors of the use and quality of group psychotherapy. The rating scale had satisfactory interrater reliability and construct validity. The immediate constructive educational critique given by the observers of the therapy groups was highly valued by group therapists. CONCLUSIONS: The monitoring and evaluation program proved to be a practical, positive, and inexpensive means of assuring and improving the quality of group psychotherapy in a clinical setting. PMID- 8422083 TI - Personality disorder in patients infected with HIV: a controlled study with implications for clinical care. AB - OBJECTIVE: Previous studies of psychiatric illness in populations infected with or at risk for HIV have not included systematic evaluation for personality disorders. The authors present the first controlled study of 1) personality disorders in HIV-positive and HIV-negative homosexual men and 2) the impact of personality disorder on coping, social support, and mood in asymptomatic HIV positive homosexual men. METHOD: The authors studied 58 asymptomatic HIV-positive and 53 HIV-negative homosexual men living outside the high-prevalence epicenters of the AIDS epidemic. Personality disorder was assessed with a clinician administered interview, the Structured Clinical Interview for DSM-III-R. RESULTS: There was a significantly higher prevalence of personality disorder in the HIV positive (33%) than in the HIV-negative (15%) subjects. In the HIV-positive subjects, those with a personality disorder (compared to those without a personality disorder) showed 1) significantly greater mood disturbance, with higher scores on the Hamilton Rating Scale for Depression, Hamilton Rating Scale for Anxiety, Profile of Mood States Total Mood Dysfunction, and the Beck Hopelessness Scale, 2) greater use of denial and helplessness when coping with the threat of AIDS, and 3) greater social conflict. CONCLUSIONS: These findings suggest that personality disorder is common in the HIV-positive population. Compared with HIV-infected patients without a personality disorder, patients with a personality disorder may experience greater dysphoria and be more likely to cope with the threat of AIDS in a dysfunctional way. Recognition of the impact of personality disorder on coping with HIV infection is important for comprehensive, sensitive, and effective clinical care. PMID- 8422085 TI - A double-blind crossover pilot study of l-deprenyl (selegiline) combined with cholinesterase inhibitor in Alzheimer's disease. AB - The potential efficacy of oral l-deprenyl (5 mg b.i.d.) added to the regimen of 10 patients with Alzheimer's disease receiving either tacrine or physostigmine was assessed in a double-blind, placebo-controlled, 4-week, two-period crossover pilot study. l-Deprenyl was associated with significant improvement in scores on the cognitive subscale of the Alzheimer's Disease Assessment Scale, suggesting possible additive effects of l-deprenyl to the effects of cholinesterase inhibitors. PMID- 8422086 TI - HIV-related risk behaviors among psychiatrically hospitalized adolescents and school-based adolescents. AB - The authors compared the responses of 76 adolescents on an inpatient psychiatric service with those of 802 school-based adolescents in the same city regarding HIV risk behaviors. The psychiatrically hospitalized adolescents reported a significantly higher rate of sexual and drug-related behaviors that involve a risk for contracting sexually transmitted disease, including HIV-related illnesses. PMID- 8422087 TI - Manic syndrome early and late in the course of HIV. AB - In a chart review at a hospital's infectious disease AIDS clinic, manic syndromes affected 8% of patients who had AIDS. Of the 14 patients with manic episodes, those without a family or personal history of mood disorder presented later in the course of HIV infection and had a higher prevalence of comorbid dementia. PMID- 8422088 TI - Treatment of alcoholism among schizophrenic outpatients: 4-year outcomes. AB - In this pilot study, the authors assessed 4-year outcomes for 18 schizophrenic outpatients with alcoholism who were treated in an innovative dual-diagnosis program. Over half (61.1%) achieved stable remissions from alcoholism. The mean duration of remission was 26.5 months. PMID- 8422089 TI - Correlation of severity of psychiatric patients' delusions with right hemispatial inattention (left-turning behavior). AB - Studies associate psychotic disorders with various forms of subtle inattention to the right hemispace (left-turning behavior). The authors examined the correlation between this dopamine-related sign and severity of delusions (presumably dopaminergic symptoms) in 20 psychotic patients. Delusions were significantly correlated with severity of left-turning bias, and this neurological sign accounted for 33% of the variance in severity of delusions. PMID- 8422090 TI - Correlation between antisaccade and Wisconsin Card Sorting Test performance in schizophrenia. AB - In 27 patients with chronic schizophrenia, there was a significant correlation between performance on an antisaccade eye movement task and on the Wisconsin Card Sorting Test. A significant correlation was not obtained between antisaccade task performance and scores on the modified Mini-Mental State examination or the Schedule for the Assessment of Negative Symptoms in Schizophrenia. In addition, patients' antisaccade task performance was impaired compared with that of 12 normal subjects. PMID- 8422091 TI - Blunted growth hormone responses to growth hormone-releasing factor and to clonidine in panic disorder. AB - Blunted growth hormone (GH) responses to growth hormone-releasing factor (GH-RF) and clonidine have been reported in patients with panic disorder. In this study GH-RF and clonidine were administered to 13 patients with panic disorder and 20 healthy volunteers. Compared to the normal subjects, the patients with panic disorder had significantly blunted GH responses after both GH-RF and clonidine. PMID- 8422092 TI - Extrapyramidal symptoms and cocaine abuse. PMID- 8422093 TI - Heroin withdrawal precipitated by nonmedical use of naltrexone. PMID- 8422094 TI - Domestic violence associated with anabolic steroid abuse. PMID- 8422095 TI - Relapse in a clozapine responder following lorazepam withdrawal. PMID- 8422096 TI - Psychotropic medications and priapism. PMID- 8422097 TI - Sleep deprivation in an elderly man with Parkinson's disease. PMID- 8422098 TI - Failed suicide or successful male genital self-amputation? PMID- 8422099 TI - Driving and psychiatric illness. PMID- 8422100 TI - Update on the dementia spectrum of depression. PMID- 8422101 TI - Pain in depression and Parkinson's disease. PMID- 8422102 TI - Diagnostic criteria for multiple personality disorder. PMID- 8422103 TI - Treatment of stuttering with phenelzine. PMID- 8422104 TI - Seasonal patterns of bulimia nervosa. PMID- 8422105 TI - Is childhood sexual abuse a risk factor for bulimia? PMID- 8422107 TI - Effects of pill-giving on maintenance of placebo response. PMID- 8422106 TI - Alprazolam in the emergency treatment of schizophrenia. PMID- 8422108 TI - Cutaneous manifestations of histoplasmosis in the acquired immune deficiency syndrome. AB - The clinical and histologic features of cutaneous histoplasmosis in three patients with acquired immunodeficiency syndrome (AIDS) are described. The patients presented with multiple discrete papules on the extremities, trunk, and face, some of which were follicular. Histologically, the skin biopsies were characterized by a sparse perivascular infiltrate with polymorphonuclear leukocytes, lymphocytes, and occasional histiocytes. Prominent leukocytoclasia and associated dermal necrosis were seen around the superficial blood vessels of the dermis. The Histoplasma capsulatum organisms were for the most part extracellular and difficult to visualize on the hematoxylin and eosin-stained sections. A diagnosis of atypical leukocytoclastic vasculitis was considered. Histoplasmosis is a relatively common mycosis among AIDS patients, and it is sometimes the first manifestation of the syndrome. The clinical and histologic findings described herein may be relatively common among AIDS patients and are quite different from those of classic disseminated histoplasmosis. PMID- 8422109 TI - Proliferating cell nuclear antigen (cyclin) expression in normal and abnormal cervical squamous epithelia. AB - Expression of Proliferating cell nuclear antigen (PCNA) was evaluated in formalin fixed, paraffin-embedded normal and abnormal cervical squamous epithelia using immunoperoxidase stains and PC10 monoclonal antibody to PCNA. PCNA was exclusively expressed in the parabasal and basal layers of normal ectocervix and a similar pattern was seen in nine of the 11 cases with squamous metaplasia. Examples of cervical dysplasia showed expression in higher layers of cervical epithelium, corresponding to the degree of dysplasia. Increased staining was seen in condylomas and markedly reduced staining with atrophy. The percentage of basal cells that stained increased progressively from atrophic to normal, to condylomatous, to dysplastic epithelia. Proliferative activity can be satisfactorily assessed in formalin-fixed cervical epithelia using PC10 PCNA antibody. This assessment can be of potential diagnostic use in difficult cases. PMID- 8422110 TI - Hodgkin's disease following non-Hodgkin's lymphoma. A clinicopathologic and immunophenotypic study of nine cases. AB - We describe nine patients who initially developed non-Hodgkin's lymphoma and subsequently developed Hodgkin's disease. The median interval from the diagnosis of non-Hodgkin's lymphoma (NHL) to the diagnosis of Hodgkin's disease (HD) was 5 years (range, 2-12 years). The median age of the patients at time of diagnosis of NHL was 54 years (range, 27-81 years). Seven of nine cases (78%) of NHL were primarily nodal. According to the Working Formulation, seven NHL were follicular (two small cleaved cell, three mixed small and large cell, two large cell), one was diffuse large cell, and one was large cell immunoblastic. All NHL had histologic or immunophenotypic findings indicative of B-cell lineage. Seven of the nine patients were treated in a nonuniform manner: four with chemotherapy and three with chemotherapy and radiation therapy. At the time of HD, the median age of the patients was 59 years (range, 35-85 years). Lymph nodes were involved in all patients. Six HD biopsies were subclassified as nodular sclerosis, one as mixed cellularity, and two cases were not further subclassified. Immunophenotypic studies revealed that the Reed-Sternberg and Hodgkin cells were LeuM1 or BerH2 positive and LCA negative in eight of nine biopsies, supporting the histologic diagnosis. These results further demonstrate that patients with NHL may subsequently develop HD. The NHLs are usually of B-cell lineage. The results also emphasize the need for rebiopsy in patients with NHL who experience an apparent clinical relapse. PMID- 8422111 TI - Malignant lymphomas involving the ovary. A clinicopathologic analysis of 39 cases. AB - We report 39 patients with non-Hodgkin's lymphoma who initially presented with ovarian enlargement. Fifteen patients had unilateral (10 left, four right, one unknown) and 24 had bilateral ovarian masses. The median size of the tumors was 8 cm (range, 2 to 23 cm). Histologically, using the Working Formulation, 21 lesions (54%) were classified as small noncleaved cell, Burkitt's type; 12 (31%) as large cell (nine diffuse, three focally follicular); three (8%) as diffuse mixed, small and large cell; two (5%) as large cell immunoblastic; and one (2%) as follicular and diffuse small cleaved cell. Twenty-six tumors were analyzed immunophenotypically, 25 (96%) of which were B-cell neoplasms. However, combining histologic and immunohistochemical findings, 37 neoplasms were of B-cell lineage, one diffuse large-cell lymphoma was not analyzed, and one large-cell immunoblastic lymphoma (with features of anaplastic large-cell lymphoma) was of T cell lineage with an aberrant immunophenotype. On the basis of staging studies and clinical follow-up, we conclude that only four (10%) of the neoplasms in this study most likely arose in the ovary. The primary neoplasms, three diffuse large cell and one diffuse mixed small- and large-cell, were B-cell neoplasms. Three of four patients with primary neoplasms were apparently cured at last follow-up following surgical excision and chemotherapy. The remainder of the lymphomas in this study, most commonly of small noncleaved cell, Burkitt's type, appear to be systemic tumors that involved the ovaries secondarily. Approximately 40% of patients with systemic neoplasms were alive without evidence of disease at last follow-up. PMID- 8422112 TI - Adenocarcinoma of the colon simulating primary urinary bladder neoplasia. A report of nine cases. AB - Nine cases of adenocarcinoma of the colon, secondarily involving the urinary bladder mucosa and histologically mimicking primary bladder neoplasia, are reported. Five patients presented with bladder involvement at the time of diagnosis of colon cancer; four developed vesical lesions 9 to 66 months after resection of their colonic primary. The majority (89%) had genitourinary symptoms at presentation; gastrointestinal manifestations were present in only 60% of those with synchronous colonic involvement. The initial clinical impression, largely based on cystoscopic and radiographic studies, was a bladder primary in four cases and colon cancer in five. Of the former, three (75%) were known to have a history of colon cancer. Histologically, all were enteric-type adenocarcinomas and all had features mimicking a villous adenoma of the bladder. Distinguishing a primary bladder adenocarcinoma from spread of a colonic carcinoma to the bladder may not be possible on histopathologic grounds alone. Consideration should be given to the possibility of an extravesical primary even when symptomatology, cystoscopy, radiographic studies, and histopathology suggest a primary bladder neoplasm. PMID- 8422113 TI - Biliary adenofibroma. A heretofore unrecognized benign biliary tumor of the liver. AB - Benign biliary tumors of the liver are uncommon. In this report, we describe a distinctive biliary tumor of 7-cm diameter occurring in the right lobe of the liver of a 74-year-old Chinese woman. The lesion, characterized by a complex tubulocystic nonmucin secreting biliary epithelial and an abundant fibroblastic stromal components, is distinct from other well-recognized biliary lesions. A number of unusual features are focally present, namely, intraluminal bile concretions, apocrine-like epithelial change, acute inflammation, and granuloma. The tumor shows a striking resemblance to Meyenburg's complex (MC), but the large size of the lesion and the absence of any typical MC in the background liver are exceptional for the latter. Its expansile growth, possession of mitoses, and foci of epithelial tufting and cellular atypia favor a neoplastic process. Previous reported cases of adenomatous neoplastic transformation of MC are dissimilar. We therefore conclude that this is a hitherto unrecognized biliary tumor that may be yet another neoplastic form of MC and propose the designation biliary adenofibroma. The course appears benign, but malignant epithelial transformation may supervene if the lesion is left untreated. PMID- 8422114 TI - Ovarian carcinoma metastasis to the breast case report and review of the literature. AB - Metastasis to the breast from extramammary malignancies is rare, but its recognition is important because the prognosis and treatment differ from that of primary breast cancer. We report a case of ovarian cancer with metastasis to the breast, which was found at the time of presentation. A 57-year-old woman presented with shortness of breath and was found to have a malignant pleural effusion. A right breast nodule contained papillary adenocarcinoma. Laparotomy showed bilateral ovarian papillary cystadenocarcinoma with dissemination in the peritoneal cavity. DNA image analysis showed multiple aneuploid stem lines. Immunohistochemical staining was positive with ovarian tumor marker OC125 but negative with breast tumor marker gross cystic disease fluid protein-15 (GCDFP 15) and estrogen receptor. The breast specimen was positive with OV632, a more specific tumor marker for ovarian cancer, thus favoring the ovary as the site of the primary tumor. PMID- 8422115 TI - Intraosseous lipoma. PMID- 8422116 TI - Microglandular adenosis, apocrine adenosis, and tubular carcinoma of the breast. An immunohistochemical comparison. AB - Four cases of microglandular adenosis (MA), together with four cases of apocrine adenosis (AA) and 10 cases of tubular carcinoma (TC) of the breast were studied at the light and immunohistochemical level. One case of MA was studied with electron microscopy. MA is characterized by an absence of myoepithelial cells (ME), epithelial membrane antigen (EMA), and gross cystic disease fluid protein (GCDFP-15). The absence of EMA in MA makes it unique among benign glandular hyperplasias of the breast. AA contains myoepithelial cells and a distinct basal lamina. It is characterized by the presence of GCDFP-15, the specific apocrine marker, which is not present in MA. TC lacks both myoepithelial cells and a basal lamina. It is negative for GCDFP-15. Periductal and vascular elastosis are common and usually prominent, whereas they are not found in either MA and AA. Other stromal changes further distinguish the three lesions. These three distinct entities can be separated objectively and unequivocally and it is essential that this be done so as to prevent confusion. PMID- 8422117 TI - In concept and practice: quality in nursing care. PMID- 8422118 TI - ANA letter to Clinton: first 100 days are critical. Interview by Mandy Mikulencak. PMID- 8422120 TI - "Juggling act" a daily routine for nurse researcher. PMID- 8422119 TI - Long-term care serves more than just elders. PMID- 8422121 TI - Improving quality through clinical practice guidelines. PMID- 8422122 TI - Cultural diversity promoter through "Ladders" program. PMID- 8422123 TI - Integrating computers and nursing care. Interview by Mandy Mikulencak. PMID- 8422124 TI - Archiving preserves nursing's heritage. PMID- 8422125 TI - HHS honors nurses on World AIDS Day. PMID- 8422126 TI - Nurses and professional boundaries: legal barriers to practice. PMID- 8422127 TI - A crying need for immunization. PMID- 8422128 TI - Exceeding consumer expectations in health care. PMID- 8422129 TI - Share ideas to speed CQI success. As I see it. PMID- 8422130 TI - Reforming health care, transforming nursing. PMID- 8422131 TI - Coalitions bring quality to health care reform. PMID- 8422132 TI - Exploring the legal aspects of quality improvement. PMID- 8422133 TI - Merging theory and reality: three quality programs in action. PMID- 8422134 TI - Congress of Nursing Practice discusses malpractice reform, payment to NPs. PMID- 8422135 TI - Congress on Nursing Economics addresses payment reform, UAP. PMID- 8422136 TI - Surgical face masks: protection of self or patient? PMID- 8422137 TI - Management of screen-detected breast cancer: audit of the first 100 cases in the Southampton and Salisbury breast screening programme. AB - With the natural history and optimal treatment of a high proportion of screen detected breast cancers yet to be determined, treatment poses the management team with a number of therapeutic dilemmas. This study surveys the management policy and treatment of a consecutive series of 100 screen-detected cancers treated in a single breast unit. The problems encountered are discussed. There were 87 women with stage Tis or T1 tumours, including 26 women with in situ cancers, four with invasive cancers less than 5 mm in size, and seven with tubular cancers. Sixty six women were managed with breast-conserving surgery and 36 women underwent localisation biopsy as the sole surgical treatment of the breast. With selection bias for high-grade and lateral tumours, only 2/13 cancers up to 10 mm in size were lymph node positive on axillary clearance. All lymph node positive women received adjuvant therapy. No adjuvant therapy was given in 43 cases, including those with in situ cancer. Thirty-six had extensive intraductal component (EIC). Patient and surgeon choice tends to be a major factor both in type of surgery and adjuvant therapy for screen-detected breast cancer. The optimal treatment for tumours detected by breast cancer screening is debatable and randomised trials on their management need to be expedited. PMID- 8422138 TI - Breast size and prognosis in early breast cancer. AB - The influence of breast size on the prognosis of 196 patients with early breast cancer diagnosed in the period 1984-1985 was studied. Breast size was based on the volume from mammography. This method was validated against the volume of the mastectomy specimen determined by water displacement in 18 patients and found to be accurate (r = 0.93, P < 0.01). The median breast volume was 833.5 cm3 (interquartile range 522.8-1153.3 cm3). Breast size was significantly associated independently with age (Spearman's rank r = 0.24. P = 0.001), menstrual status (z = -4.81, P < 0.001), body weight (Spearman's rank r = 0.61, P < 0.001), T stage (z = -1.91, P = 0.05) but not N stage (z = -1.64, P = 0.10) or hormone receptor status (z = -0.80, P = 0.42). In an analysis of breast size and other known prognostic factors, based upon Cox's proportional hazards regression, N stage was the only significant factor for both breast cancer survival and disease-free survival. Even though women with larger tumours at presentation had larger breasts, breast size was not a significant prognostic factor in early breast cancer. PMID- 8422139 TI - Should we still stitch the subcutaneous fat layer? A clinical and ultrasound assessment in 50 hip operations. AB - Wound haematoma is an undesirable complication of surgery. We report a prospective trial to establish whether a subcutaneous fat stitch affects haematoma formation in hip surgery. A series of 50 patients undergoing hip surgery were randomised to have either a fat stitch or no fat stitch during wound closure. The wounds were assessed clinically with a minimum follow-up of 5 weeks. Subcutaneous haematomas were seen more easily with the patient standing. As clinical assessment for haematoma formation may be unreliable, each patient also had an ultrasound scan. Although the incidence of subcutaneous haematoma was slightly higher in the no fat stitch group (36% compared with 24% in the fat stitch group) this difference was not statistically significant (chi 2 test, P < 0.5). There were no significant differences in the incidence of wound infection and healing rate. From our study we also showed that ultrasound examination was twice as sensitive as clinical examination in diagnosing subcutaneous haematomas and that subcutaneous haematomas are common after hip surgery. PMID- 8422140 TI - Comparison of 'intraoperative' parathormone measurement with frozen section during parathyroid surgery. AB - Intact parathyroid hormone (PTHi) has a relatively short half-life and levels fall significantly within 15 min of the successful excision of all the abnormal parathyroid glands during surgery for hyperparathyroidism. Monitoring this fall has been suggested as useful in decreasing the failure rate of neck explorations in parathyroid surgery. We have performed a comparative study of the use, during surgery for primary hyperparathyroidism, of routine frozen section with the recently available rapid assay for PTHi. The assay demonstrated a significant fall (P = 0.013) in the level of PTHi from a pre-excision mean of 26.66 pmol/l to 5.94 pmol/l 15 min after surgical excision. In all cases the level of PTHi fell to less than 30% of pre-excision levels by 30 min. However, while frozen section results were available in a mean time of 22 min after excision, the PTHi levels took a mean of 105 min. We conclude that during straightforward parathyroid surgery for primary hyperparathyroidism, the current assay does not offer any advantages over the already routine use of frozen section. PMID- 8422141 TI - Lipoprotein(a) and its role in occlusive vascular disease. AB - An increased serum level of lipoprotein(a) (LP(a)) is acknowledged as an independent risk factor for occlusive vascular disease, especially atherosclerosis. Recently, a considerable amount of data has been produced on the structure and functional relationships between Lp(a) and the primary cellular interactions of atherosclerosis. In this review we discuss the structure and function of Lp(a) with specific reference to its role in the proposed mechanisms of atherosclerosis. PMID- 8422142 TI - How long do patients convalescence after inguinal herniorrhaphy? Current principles and practice. AB - Over the course of this century it has become apparent that there is no longer any rationale behind the old-established advice to rest for several weeks after hernia repair. It was our impression that such advice continues to be widely accepted, and we therefore sent questionnaires to 100 recently appointed consultant surgeons, 400 of their patients and 200 recently established partners in general practice to assess current practices. Our findings show that surgeons advised a mean of 4.4 weeks off work and GPs 6.2 weeks off-work, in both cases the period varying with the nature of the patient's occupation. Patients actually took a mean of 7.0 weeks off work. The wide variation reflects the lack of evidence that an early return to work after hernia repair causes any detrimental effect. We believe that this should be explained to patients, who should be free to return to work as soon as they feel comfortable. Such a policy could substantially decrease the current loss of productivity. PMID- 8422143 TI - Infantile hypertrophic pyloric stenosis: where should it be treated? AB - A retrospective analysis in the form of an audit into the management of infantile hypertrophic pyloric stenosis in a district general hospital has revealed that the results are equivalent to that of published data from specialised units. It is stressed in this study that close co-operation has to be maintained between paediatricians and surgeons in the care of these infants. The diagnosis can be made on clinical grounds in the majority of cases. The operation has to be carried out by experienced surgeons and anaesthetists. The morbidity can be minimised under these circumstances and pyloromyotomy can be performed safely in a district general hospital. PMID- 8422144 TI - Hepatic 'pseudotumours': an important diagnostic pitfall. AB - Hepatic 'pseudotumours' are radiological abnormalities which give the appearance of tumour. They may be focal areas of disease such as fatty infiltration or areas of focal sparing in diffuse processes. We present four cases of hepatic 'pseudotumours' due to diffuse fatty infiltration with focal sparing and emphasise the need for further radiological assessment of focal lesions found on ultrasound scanning. Measurement of CT attenuation allows a confident diagnosis of fatty infiltration with focal sparing, thus preventing unnecessary investigation. Other forms of hepatic pseudotumour may require further investigation. PMID- 8422145 TI - Colostomy is no longer appropriate in the management of uncomplicated large bowel obstruction: true of false? PMID- 8422146 TI - Gastrointestinal complications after cardiac surgery. AB - Gastrointestinal complications after cardiac surgery are uncommon, but are associated with a high morbidity and mortality. Over 11 years 8559 procedures requiring cardiopulmonary bypass were performed in this unit and 35 patients were identified who developed gastrointestinal complications after surgery, an incidence of 0.41%. There were nine deaths in this group, a mortality of 25.7% compared with an overall mortality after cardiac surgery in Ireland ranging from 3.24% to 4.81%. These complications required surgery in 21 patients. The most common indication for surgical intervention was upper gastrointestinal bleeding in 10 patients, three patients had acute pancreatitis, two patients had perforated peptic ulcer; two patients had intestinal ischaemia, with five cases of colon pathology. The difficulties of making an early diagnosis are outlined and a low threshold to exploratory laparotomy is recommended. PMID- 8422147 TI - Can efficiency of follow-up for superficial bladder cancer be increased? AB - This study evaluated the efficiency with which follow-up cystoscopy was employed in a population-based cohort of patients with superficial bladder cancer. Subjects were 240 men, aged under 75 years, with superficial bladder cancer first diagnosed in 1982. The median duration of follow-up was 6.1 years. The median (interquartile range) number of follow-up cystoscopies was 8 (5-12) per patient with a patient-specific annual rate of 1.7 (1.2-2.2) per year. The median number of cystoscopies at which recurrent tumour was detected was 2 (0-5) per patient, patient-specific annual rate 0.4 (0.0-1.3) per year of follow-up. Among patients with a single tumour at diagnosis and a negative first check cystoscopy (MRC group 1), the proportion of positive cystoscopies was 0.1 (0.0-0.4). Comparison of observed intervals between cystoscopies with optimal intervals calculated using an optimisation model showed that patients in MRC group 1 were seen sooner in practice than the model predicted, while in practice other patients were seen later than the model predicted. These data support the suggestion that efficiency of follow-up for patients with superficial bladder cancer could be increased by dividing patients into groups with different risks of recurrence and differing follow-up requirements. PMID- 8422148 TI - Informed consent: a case for more education of the surgical team. AB - A questionnaire was given to 37 members of staff of the Department of Surgery, Addenbrooke's Hospital, Cambridge, in order to determine whether their knowledge was adequate to give accurate information to patients regarding operations and thus to obtain properly informed consent for that operation. Each participant was asked to estimate the 24-h and 30-day mortality for five common elective operations. A wide range of answers was given for operations by all groups. Estimates of 24-h mortality after unilateral inguinal herniorrhaphy differed between staff grades by a factor of 3, but estimates of 24-h mortality after thyroidectomy differed by a factor of 100 between consultant surgeons and staff nurses. Our findings suggest that some members of the surgical team have insufficient knowledge about common operations to obtain properly informed consent from patients. PMID- 8422149 TI - Amputations in diabetics. PMID- 8422150 TI - Laparoscopic laser cholecystectomy: our first 200 patients. PMID- 8422151 TI - Can cholangiography be safely abandoned in laparoscopic cholecystectomy? PMID- 8422152 TI - Alcohol-related hand injuries: an unnecessary social and economic cost. PMID- 8422153 TI - Non-tumour morbidity and mortality after modified radical mastectomy. PMID- 8422154 TI - A study of the effects of Picolax on body weight, cardiovascular variables and haemoglobin concentration. PMID- 8422155 TI - Prevention of bacterial infection and sepsis in acute severe pancreatitis. PMID- 8422156 TI - Intestinal ischaemia in the unconscious intensive care unit patient. PMID- 8422157 TI - Blood loss and transfusion requirements in total joint arthroplasty. PMID- 8422158 TI - Emergency treatment of tracheal tear during pharyngolaryngectomy. PMID- 8422159 TI - Will screening for breast cancer reduce mortality? Evidence from the first year of screening in Avon. AB - In the first year of screening in Avon, 93 malignant lesions were detected of which one-half were impalpable. Of the impalpable lesions, one-half were in situ or showed areas of microinvasion only. One-fifth of the malignant lesions were invasive tumours of special histological type which are known to carry a good prognosis even when not detected by screening. One-quarter of the lesions had clinical or pathological features which would be expected to confer a poor prognosis. Only 16 invasive ductal carcinomas measuring 1 cm or less in diameter were detected--a small proportion of the total number of malignant lesions. Although these early figures suggest that the effect of screening on mortality from breast cancer may be small, continued high-quality screening and careful detailed analysis are essential to determine the effect of screening on the mortality from breast cancer and the effect on the population as a whole. PMID- 8422160 TI - A waiting list initiative in general surgery--experience in a large district general hospital. AB - In a waiting list initiative scheme 566 operations were performed on 447 patients in 104 sessions over a period of six months. In addition the names of 78 other patients were deleted from the waiting list. Special attention was paid to pre operative education and assessment of patients by information sheets and 'question and answer' history sheets respectively. The maximum waiting time for all general surgical patients was reduced from 2.5 years to less than one year and the total waiting list reduced from 2254 to 1529 patients. PMID- 8422161 TI - Inadequacies of hospital medical records. AB - We have assessed the extent to which hospital records follow the Guidelines for Medical Records and Notes published by the Royal College of Surgeons of England. Notes of 100 consecutive discharges were reviewed from two surgical units, one at a District General Hospital (DGH) and the other at a London Teaching Hospital (TH). Overall, only 65 per cent (DGH) and 67 per cent (TH) of the entries specified by College guidelines were both present and correct. Substandard categories included the regular update of notes, post-operative instructions, comments about post-operative recovery, the record of advice given to relatives and incorrect consent. The guidelines produced by the Royal College of Surgeons are being applied, but there is room for considerable improvement. Inadequate medical records limit audit and may have medico-legal consequences. We recommend regular assessment of the standard of note keeping. PMID- 8422162 TI - Disruption caused by the house officer's bleep: a simple solution. AB - For a 'control' week, all bleep calls a surgical house officer received were charted without the nurses' knowledge and graded according to an arbitrary scale of usefulness. Also recorded were time of day, the person bleeping and whether the house officer was involved in patient care. A system was then tried in a later 'test' week, whereby only the nurse in charge would bleep the house officer and a board was placed on the house officer's own two wards where tasks could be written. Again all calls were recorded. This led to a decrease in inappropriate bleep calls from the nurses (58 per cent falling to 15 per cent p < 0.01) and a fall in numbers of nurses' bleep calls by two-thirds before 5 pm (p < 0.01). There was also a decrease in the number of bleeps when the house officer was directly involved in patient care (46 per cent falling to 28 per cent). The nurses in charge were agreeable to the changes. The entire study was repeated by another house officer with similar results. There has been little research on the disruption caused by the bleep and we believe that by this simple policy the house officers' day can be disrupted far less. PMID- 8422163 TI - Height, weight, BMI and weight-for-height of adults in southern Iran: how should obesity be defined? AB - Height, weight, body mass index (BMI) and weight for height are reported for 2210 parents of a random sample of school children in Shiraz, Iran. Weight for age and weight for height centiles have been computed by the Healy, Rasbash and Yang's nonparametric method and are presented graphically. The Iranian adults are taller and heavier than previously reported for people in the region but are smaller than European and US white adults. The distribution of BMI is broadly similar to that reported for UK adults but curves corresponding to BMI-constant, i.e. weight proportional to height, cut across the centiles of weight for height. PMID- 8422164 TI - Growth and health status of preschool Karen highlanders. AB - The physical growth and health status of preschool (< 6 years) Karen highlanders belonging to the Pwo (212 males, 227 females) and Sgaw (107 males, 101 females) ethnic groups, and residing in northwest Thailand are described in the present report. The growth and health status of Thai tribal groups, including the Karen, is generally assumed to be substantially poorer than those of Thai lowlanders, who are primarily of Thai ancestry. However, the Karen highlanders in the present report fell within the range of variation in growth and health status found in lowland southeast Asians. Also, the growth and health status of Pwo preschool children tended to be significantly poorer than those of Sgaw children, despite the fact that both groups reside in isolated villages within the same ecozone and utilize similar food production techniques. The cause of these differences cannot be determined at the present time, but several potential factors are discussed. PMID- 8422165 TI - Pulmonary function studies in healthy non-smoking women of Calcutta. AB - Pulmonary function measurements were made in 230 healthy non-smoking women from Calcutta with an age range of 20-59 years. The tests consisted of vital capacity (VC), forced vital capacity (FVC), forced expiratory volume in one second (FEV1), forced expiratory volume in one second as a percentage of forced vital capacity (FEV1%), forced expiratory time (FET), maximum voluntary ventilation, uncontrolled frequency (MVVF), forced expiratory flow (FEF200-1200ml), forced mid expiratory flow (FEF25-75%), forced end-expiratory flow (FEF75-80%) and peak expiratory flow rate (PEFR). Except for PEFR, all the measurements were made with the help of two 9-litre closed-circuit-type expirographs using standard spirometric techniques. PEFR was recorded with the help of two Wright peak flow meters. Prediction formulae were derived on the basis of physical characteristics. Age and height were found to be the significant predictor variables for VC, FVC and FEV1, while only age was significant for FEV1%. The FVC and FEV1 values of the subjects, standardized for age and height, are much lower than those of Americans, Europeans and Jordanians. On comparison with data reported from other parts of India, it was revealed that the VC and FEV1 values of the current study, after adjustment for age and height, were much higher than those of Southern Indians but comparable with those of North-Western Indians. PMID- 8422166 TI - Serum alkaline phosphatase activity and skeletal maturation in Guatemalan adolescents. AB - Serum alkaline phosphatase activity (AP), stature and Tanner-Whitehouse-2 RUS skeletal age (SA) were determined for 873 rural Guatemalan youth and young adults 11-25 years of age. Mean AP decreases systematically with chronological age (CA) in girls, while an adolescent increase in mean AP occurs at 14 years in boys. When mean AP is calculated within SA groups, clear adolescent increases are apparent in girls, peaking at 11 years SA, and at 14 years SA in boys. Correlations between AP and relative skeletal age (SA-CA) decrease systematically from moderately positive at the youngest CA groups to moderately negative at the older ages, crossing zero at the chronological ages of maximum adolescent AP values. Analyses suggest the patterns of mean AP in adolescence follow closely the timing and patterns of growth velocity in stature in each sex, and are only indirectly related to SA as it is a proxy for the maturational timing of the adolescent spurt in stature. PMID- 8422167 TI - Anthropometric-hormonal correlation patterns in fertile and post-menopausal women from Austria. AB - Correlations between sex-hormone levels and body dimensions were investigated in a group of women from Vienna. Since sex-hormone concentrations in women are subject to dramatic changes with increasing age, both 124 fertile and 142 postmenopausal women were examined. Twenty-nine anthropometric traits were correlated with oestradiol, the gonadotrophins luteinizing hormone (LH) and follicle-stimulating hormone (FSH), the gestagens progesterone, 17 hydroxyprogesterone and prolactin, and the androgens testosterone, androstenedione and DHEA-S, as well as with the transport protein, sex-hormone binding globulin. It was also found that oestradiol concentration was related to levels of gonadotrophins, prolactin, progesterone and androgens and was equally correlated with metric traits. Statistically significant correlations could be observed between all sex-hormone concentrations and the 29 anthropometric traits. The most striking result is the change in direction of correlations between metric traits and oestrogen, gonadotrophin, prolactin and gestagen levels between both proband groups, which is probably due to changes in sex-hormone concentrations in women after menopause. The direction of correlations with androgens remained the same. In post-menopausal women the great impact of fat tissue for sufficient synthesis of oestrogen is shown by a positive correlation between oestradiol and circumference measures. PMID- 8422168 TI - Ages at peak velocity and peak velocities for seven body dimensions in Japanese children. AB - Seven body dimensions of 37 Japanese children, 16 boys and 21 girls, were measured longitudinally at 6-monthly intervals from 9 through 15 years of age. Spline curves were fitted to the longitudinal data of each dimension to obtain age at peak velocity and magnitude of peak velocity for each child. Ages at peak velocity for each dimension occurred, on average, later in boys than in girls. Mean peak velocities were also larger in boys except for chest circumference. Ages at peak velocity occurred, on average, earlier in Japanese than in European and North American children. PMID- 8422169 TI - Pubertal development in south-Hungarian boys and girls. AB - Data on pubertal development in 1392 boys and 1494 girls from the cross-sectional Pecs Growth Study are reported. Genital, breast and pubic-hair development were rated according to Tanner (1962). The initiation of pubic-hair development in boys was 11.8 years and that of genital development was 12.0 years. Stage 2 of pubic-hair development (PH2) in boys occurred 1.7 years later than in girls. The mean time of different stages of pubertal development in boys was in the same range as in Eveleth and Tanner's study (1991). Stages 2, 3 and 4 for pubic-hair and breast development in Pecs girls occurred earlier than in Eveleth and Tanner's subjects (1991), while the mean time of stage 5 for both sexual characteristics occurred at a later age than reported in Eveleth and Tanner (1991). The age of menarche was 12.9 years in Pecs girls, which seems later by 0.1-0.6 years than in Western-European countries and earlier by 0.1-0.3 years than in South-European countries. PMID- 8422171 TI - List of members of the Society for the Study of Human Biology. PMID- 8422170 TI - Reference values and standards for paired organs. AB - When estimating reference values for measurements on paired organs, the standard deviation for a single measurement can be obtained to a good approximation simply by ignoring the pairing. In practice, however, there are advantages in the use of conventional standards for pair means and differences. PMID- 8422172 TI - [Development of fluorinated pyrimidines in Japan]. AB - Four fluorinated pyrimidines that are in the stage of clinical trials at present in Japan were reviewed: BOF-A2, Ro09-1390, TT-62 and S-1. Both BOF-A 2 and S-1 are a compound of 5-FU derivative combined or mixed with an inhibitor of 5-FU degradation in order to prolong the blood 5-FU level as well as increase selective toxicity to tumor. Furthermore, an inhibitor of 5-FU phosphorylation in G1 tract contained in S-1 reduces G1 toxicity such as diarrhea etc due to prolongation of blood 5-FU level. Ro09-1390 is an improved compound of 5'-DFUR, which intends to reduce diarrhea caused by the latter. TT-62 is a FdUMP derivative and an active metabolite of 5-FU for oral formulation, which is superior to available 5-FU type anticancer agents in efficacy, and doesn't show cross tolerance to 5-FU. PMID- 8422173 TI - [Cooperative Study of Surgical Adjuvant Chemotherapy for Colorectal Cancer (third report): five-year results after surgery]. AB - Beginning in January 1984, in collaboration with 357 institutions nationwide, comparative studies on curative resection cases of colorectal cancer and a control group were conducted for 2 years. The following method of administration was employed; Group A received 12 mg/m2 of MMC on the day of operation, followed by 6 mg/m2 every 2 months 6 times. From the 2nd week after the operation, 800 mg/body/day of Futraful was administered for one year. Group B received no treatment. Altogether 2786 cases were collected and, excluding ineligible cases, 2477 evaluable cases were analyzed. There was no difference in either survival rate or disease free rate, but after the bias of background factors was corrected by the hazard model, the disease free rate was better with Group A, and there was a significant difference with the rectum. According to the Dukes classification, in the Dukes C cases with the rectum Group A had higher rate of survival, disease free rate and lower rate of metastasis to the liver and local recurrence of cancer. PMID- 8422174 TI - [Cooperative Study of Surgical Adjuvant Chemotherapy for Colorectal Cancer (fourth report): five-year results after surgery]. AB - In collaboration with 428 institutions nation-wide, comparative studies on the optimal daily dosage of UFT were carried out for non-curative resection cases of colorectal cancer for two years from January 1984 to December 1985. The administration method employed was as follows. Group C received 12 mg/m2 of MMC on the day of operation, followed by 6 mg/m2 every 2 months 6 times, and from the 2nd week after the operation 600 mg/body of UFT was continuously administered every day for 1 year. Group D received 400 mg/body of UFT every day and the same amount of MMC as Group C. Analysis was made of 556 evaluable cases. There was no difference in the survival rate between the two groups. With respect to the dosage per body weight (kg), 8-12 mg/kg of UFT produced good prognoses and less side effects than the group which received more than 12 mg/kg. It seems necessary to administer UFT at its optimal dosage for colorectal cancers. PMID- 8422175 TI - [Combination therapy of high dose doxifluridine (5'-DFUR)+mitomycin C (MMC) for advanced or recurrent colorectal cancer. Joint Research Group in the Osaka Area for Combination Therapy of 5'-DFUR with Other Drugs]. AB - A co-study was conducted on treatment with high dose 5'DFUR+MMC in order to determine its effectiveness as a systemic chemotherapy in patients with advanced and recurrent colorectal cancers. The treatment schedule included 5'-DFUR given orally at a dose of 1,600 mg/body/day for five consecutive days, a two-day discontinuation, then repeated medication with the above, and MMC injected intravenously at a dose of 6 mg/m2 once every four weeks. One course consisted of four weeks, and courses were repeated as long as tolerated by the patients. Twenty-nine patients were registered for the study and 28 cases were perfectly capable of anti-tumor responses. The response rate was 25.5%, including: CR, 2; PR, 5; NC, 11; and PD, 10 cases. Responding cases were found in metastases to the lung, cervical lymph node and abdominal wall. Response rate was especially high in pulmonary metastases at 41.7% (5/12 cases). The slight side effects were mostly gastrointestinal in nature and did not necessitate drug discontinuance. The results suggest that treatment with high dose 5'-DFUR+MMC is useful for advanced and recurrent colorectal cancers, especially those with pulmonary metastases. PMID- 8422176 TI - [Studies on tissue concentration of tegafur, 5-fluorouracil, uracil after UFT administration together with the study of microangiography of colorectal cancer]. AB - In order to elucidate the effect of tumor vascularity on a various regimens concentration in tumor tissue, correlation among tegafur, 5-fluorouracil (5-FU), uracil concentrations in tissue and the microangiography were examined in 27 patients with colorectal cancer after preoperative administration of UFT (400 mg/day for 7 days). The concentrations of tegafur, 5-FU and uracil in tumor were higher than those in normal tissue (p < 0.01). There was no definite correlation between tissue concentration in each regimen and the extra or intramural vascular changes. A significant high concentration of 5-FU was found in the protuberant cancer and the ulcerative cancer with horizontal growth than that in the ulcerative cancer with vertical growth (p < 0.05). The present study indicates that the vascular changes within a tumor and the vascular pattern of tumor may be a contributing factor affecting the tissue concentration in the regimens. PMID- 8422177 TI - [Concentration of mitomycin C in portal blood and peripheral blood after intra venous, intraperitoneal or intrapelvic administration]. AB - Immediately before closing the abdomen in a curative operation for colorectal cancer, 20 mg of Mitomycin C (MMC) was administered by intravenous, intraperitoneal or intrapelvic routes, and portal blood and peripheral blood were simultaneously collected to investigate the disposition of MMC. After intravenous administration, the MMC concentration in portal and peripheral bloods changed at the same levels, showing a biphasic pattern. The maximum concentration (Cmax) seen shortly after administration was about 2 micrograms/ml. The blood halflife in phase alpha was about 5 min, and that in phase beta about 30 min. After intraperitoneal administration, the MMC concentration in portal blood was over 60 min higher than that in peripheral blood. Cmax in portal blood, seen within 5 min after administration, was about 0.5 micrograms/ml, and that in peripheral blood was about 0.2 micrograms/ml around 10 min. The half-life was about 30 min in both types of blood. After intrapelvic administration, the MMC concentration in portal blood changed at higher levels than in peripheral blood, as after intraperitoneal administration. The Cmax in portal blood, obtained within about 5 min, was about 0.3 micrograms/ml, and that in peripheral blood was about 0.1 micrograms/ml at 12 min. The half-life was about 30 min in both types of blood. As described above, the disposition of MMC was investigated on the basis of the concentration in portal and peripheral blood. As a result, intravenous MMC showed a similar change, whereas intraperitoneal or intrapelvic MMC was higher in portal blood than in peripheral blood, indicating that the disposition of MMC was different with the route of administration. PMID- 8422178 TI - [A case of secondary leukemia induced by chemotherapy with a CDDP-based regimen for gastric cancer 5 years following radical resection]. AB - Cisplatin, mitomycin C and 5-fluorouracil were given a 55-year-old woman for an unresectable gastric cancer, and successful radical gastrectomy was performed. Postoperative adjuvant immunochemotherapy using UFT and PSK was continued for about 4 years and 4 months. Pancytopenia was observed at 5 years after the treatment and then marked leucocytosis was noted. She also showed complications of general fatigue, appetite loss etc. A secondary acute leukemia associated with eosinophilia was diagnosed by peripheral blood examinations, showing WBC, 122,400: blast, 37.5 % and eosinophil, 41%. Results also showed atypia and pseudo Pelger nuclear abnormality of eosinophil, high positive stain of cell myelogenic cell surface marker, many numeral and structural abnormalities of chromosomal analysis, etc. From the above results, it was suggested that the leukemia might be induced by previously performed chemotherapy. The patient died about 2 months following its onset. PMID- 8422179 TI - [A case of bilateral pulmonary metastasis from rectal cancer successfully treated with resection after high-dose 5'-DFUR plus MMC combination chemotherapy]. AB - We reported a case of successful treatment of bilateral pulmonary metastasis from rectal cancer with high-dose 5'-DFUR plus MMC combination chemotherapy. A woman born in 1948 showed a recurrence in the bilateral lung about 29 months after low anterior resection. High-dose 5'-DFUR plus MMC combination chemotherapy was started in March, 1991. The chest X-ray examination 8 weeks after beginning this therapy showed a remarkable decrease in the size of the pulmonary metastatic foci and CEA decreased in the same way. The dose of 5'-DFUR was reduced after 5 courses, and then CEA increased. No remarkable side effect was encountered and the patient could be safely treated at an outpatient clinic. During this therapy no recurrence has been detected, and we performed a resection of the bilateral pulmonary metastasis by median sternotomy in October, 1991. The above findings suggested that this was an effective and safe therapy for pulmonary metastasis from colon cancers and could be a neo-adjuvant chemotherapy for surgical resection of pulmonary metastasis. PMID- 8422180 TI - [A case of ALL complicated with acute pancreatitis and pancreatic pseudocyst caused by L-asparaginase ]. AB - We reported a case of ALL complicated with acute pancreatitis caused by L asparaginase (L-Asp). The patient was a 42-year-old man, who showed eosinophilia in peripheral blood and an increase of lymphoblast in bone marrow. He was diagnosed as ALL (L2) and treated by JALSG '87 protocol. Remission induction chemotherapy including L-Asp was administered by 5,000 IU i.v. for 10 days. The day after giving all dose of L-Asp, slight epigastralgia developed and then became severe. After two days, s-amylase was markedly elevated, and the patient was diagnosed as acute pancreatitis caused by L-Asp. He was treated conservatively, but hyperglycemia occurred. The epigastrial tumor was palpable and gradually grew in size. CT-scan and abdominal ultrasonography revealed pancreatic pseudocyst, so he was treated by percutaneous cyst drainage. The patient died of a relapse of ALL. The prophylaxis and early diagnosis of the pancreatitis and hyperglycemia caused by L-Asp are very difficult. We have to examine more cases and pay greater attention to the chemotherapy, including L Asp. PMID- 8422181 TI - [A case of B-chronic lymphocytic leukemia/prolymphocytic leukemia (CLL/PL)]. AB - A 59-year-old man was admitted to our hospital on May 17, 1991 because of dizziness and a sense of abdominal fullness. Physical examination on admission showed splenomegaly without hepatomegaly or lymphadenopathy, and blood examination revealed normocytic anemia, thrombocytopenia and marked leukocytosis of 16,800/microliters with 87% lymphoid cells. Prolymphocytoid cells formed 28% of the lymphoid cells. Bone marrow aspiration revealed massive infiltration of lymphoid cells. Surface marker analysis showed that the lymphoid cells were positive for anti-HLA-DR, CD 5, CD19, CD20, CD21, SmIgM and SmIgD. The patient was diagnosed as having B-CLL/PL, according to the classification advocated by Melo in 1986, and initially treated with vindesine + prednisolone + pirarubicin (VP-THP). However, the prolymphocyte count increased, so we changed to VP-THP + cyclophosphamide (VEP-THP), and remission was obtained. CLL/PL is a rare disease in Japan but we obtained a good response to chemotherapy. PMID- 8422182 TI - [Anaerobic glycolysis in the culture of rat C6-glioma cells]. PMID- 8422183 TI - [Combined mitoxantrone therapy for heavily pretreated metastatic breast cancer]. PMID- 8422184 TI - [Development of antimetabolites, II]. AB - Two analogues of cytosine arabinoside (ara-C) and three analogues of methotrexate (MTX) are under their clinical studies in Japan. Gemcitabine and DMDC are analogues of ara-C, showing similar antitumor efficacies each other and rather mild animal toxicities. Currently, dFdC is under phase II study, showing activity against lung cancer and DMDC under phase I study. As for MTX analogues, edatrexate, trimetrexate and TNP-351 have been put on the clinical phase studies. Among these antifolates, both edatrexate and trimetrexate have showed responses against lung cancer and we expect that TNP-351 may also show a similar response in the future. Currently, phase II studies of edatrexate and trimetrexate and phase I study of TNP-351 are under progress. PMID- 8422185 TI - [New anthracyclines]. AB - Idarubicin showed the superior activity against Acute Non-Lymphocytic Leukemia (ANLL) in the prospective randomized trials comparing to daunorubicin and it is judged that the analog will become the first choice in the treatment on ANLL. Anthracyclines including SM-5887, KRN-8602, ME-2303 under studies in Japan have shown comparable or superior antitumor activities and lower cardiac toxicities compared to doxorubicin in preclinical studies and therefore the results obtained in clinical trials are expected. Phase II trials of anthrapyrazoles which is an analog of mitoxantrone are in progress. Among three compounds entered it is of note that CI-941 has demonstrated an excellent activity against advanced breast cancer. PMID- 8422186 TI - [Topoisomerase inhibitors developing in Japan]. AB - Irinotecan hydrochloride (CPT-11), topotecan, sobuzoxane, NC-190, and IST-622 are unique topoisomerase inhibitors and are investigational in Japan. CPT-11 is a water-soluble, semisynthetic derivative of camtothecin. CPT-11 shows its anticancer activity by inhibiting topoisomerase I activity, now a target of anticancer agents with major interest. Recent clinical trials reveal that CPT-11 is very effective in the treatment of cancer including lung cancer, cervical cancer, ovary cancer, stomach cancer, colon cancer, and non-Hodgkin's lymphoma. Major dose limiting toxicities are leukopenia and diarrhea, and are dose related. Topotecan is an another semisynthetic derivative of camtothecin and is also topoisomerase I inhibitor. Topotecan has undergone phase I clinical evaluations in USA, europe, and recently in Japan. DLF are leukopenia and neutropenia. Topotecan is more hydrophilic than its parent compound and shows lesser protein binding. Renal excretion appears to be the major route of elimination. Sobuzoxane (MST-16) is a unique derivative of dioxopiperazine, an inhibitor of topoisomerase II. In phase II studies, definite anticancer effects are observed in patients with non-Hodgkin's lymphoma and adult T-cell leukemia/lymphoma. Responses are seen even in pretreated cases. Leukopenia is also dose-limiting. Non-hematologic toxicities are mild and include alopecia and G.I. toxicities. NC-190 is a novel benzophenazine derivative with excellent antitumor activities against murine tumors. NC-190 also inhibits topoisomerase II. Now the drug is an early clinical phase II studies in Japan. Toxicities include bone marrow suppression, transient mild to moderate liver enzyme elevation, alopecia and mild G.I. toxicities. Tumor responses are occasionally encountered. IST-622 is a semisynthetic derivative of chartreusin. The drug is an inhibitor of topoisomerase II (and I in high concentration). IST-622 shows excellent, broad anticancer activity against murine tumors. The drug is well absorbed from small intestine. IST-622 is now in phase I clinical trial in Japan. PMID- 8422187 TI - [Antitumor activity of navelbine (vinorelbine ditartrate), a new vinca alkaloid analog]. AB - The antitumor activity of navelbine (vinorelbine ditartrate, KW-2307) against murine and human transplantable tumors was compared with those of other vinca alkaloids, vindesine (VDS), vincristine (VCR) or vinblastine (VLB). KW-2307 and VDS increased the life span of mice bearing ascitic tumors (P 388 leukemia, L 1210 leukemia, EL-4 lymphoma, Colon 26, FM 3 A mammary carcinoma and M 5076 sarcoma) slightly more than VCR or VLB. A significant difference was not found in the antitumor activities against 6 murine solid tumors (B 16 melanoma, Colon 26, FM 3 A mammary carcinoma, Lewis lung carcinoma, M 5076 sarcoma and Sarcoma 180). However, a remarkable difference was observed in the antitumor activities against 11 human tumors inoculated into nude mice. The activity of KW-2307 was more than those of other 3 drugs against 4 human non-small cell lung carcinomas (Lu-65, Lu 99, LC-6 and L-27) and 2 stomach carcinomas (St-4 and St-40). KW-2307 and VDS were also effective in inhibiting the growth of 2 human breast carcinomas (MX-1 and Br-10). PMID- 8422188 TI - [Phase I Study of RA-700. RA-700 Clinical Study Group]. AB - An antitumor substance, RA-700, isolated from Rubia akane or Rubia cordifolia has the novel structure. Phase I clinical study was conducted by the RA-700 clinical study group consisting of 6 institutions. A single dose administration and 5-day schedule administration were evaluated with 14 patients respectively. RA-700 was given from 0.2 to 1.4 mg/m2 in single i.v. dose study, from 0.4 to 2.0 mg/m2 in 5 day i.v. schedule study. Nausea and vomiting, fever, stomachache, mild hypotension and slight abnormality of electric-cardiogram were observed as the toxicities. In pharmacokinetic study, the elimination half-lives (t1/2) of RA-700 in plasma were 55 min, of alpha-phase and 3.9 hrs. of beta-phase by single dose study, and 23-25 min. of alpha-phase and 6-14 hrs. of beta-phase by 5-day schedule study. Accumulation was not found by 5-day schedule administration, and metabolite were not observed in plasma and urine. It seems that RA-700 is metabolized by the liver and excreted in the feces. In conclusion, the maximum tolerated dose was 1.4 mg/m2 for 5-day schedule administration. PMID- 8422189 TI - [Phase II study of NK 622 (toremifene citrate) in advanced breast cancer, a multicentral cooperative dose finding study]. AB - In order to determine the usual dose in the first line therapy and a high dose in the second or third line therapy, a dose finding study of a novel antiestrogen NK 622 (toremifene citrate) was performed in patients (pts) with advanced or recurrent breast cancer. NK 622 was orally administered daily once for more than 8 weeks. In pts without previous drug therapy or in pts with cancer relapse after adjuvant therapy, the response rates [(CR + PR)/total] were 24.1% (7/29), 13.8% (4/29), 20.0% (1/5) and 40.0% (2/5) at doses of 40, 60, 120 and 240 mg/day, respectively. A 40 mg/day dose showed an objective response only in postmenopausal pts with estrogen receptor (ER) positive or unknown cancer. At a dose of 60 mg/day, some of the responding cases were premenopausal pts or pts with ER(-) cancer. In pts with cancer relapse during adjuvant therapy or in those with previous therapy and/or radiation, response rates were 25.0% (2/8), 0% (0/4), 13.5% (5/37) and 10.3% (4/39) at doses of 40, 60, 120 and 240 mg/day, respectively. Response was more frequent in pts with ER (+) cancer than with ER ( ) cancer. The response rates in pts with previous therapy including tamoxifen (TAM) except medroxyprogesterone (MPA) were 14.3% (4/28) at a 120 mg/day dose and 6.1% (2/33) at a 240 mg/day dose. In pts with previous therapy including TAM, MPA and other antitumor agents, the rate was 18.2% (2/11) at a 120 mg/day dose. Side effects such as elevation of GOT, GPT and serum Ca level, decrease of hemoglobin, anorexia, nausea/vomiting, fatigue, dizziness and hot flush were observed. These side effects were moderate in grade and reversible. Dose dependency of side effects was not clearly observed in grade and incidence. From these results, NK 622 is expected to be a safe drug with efficacy in first line therapy at a dose of 40 mg/day and in second or third line therapy at a dose of 120 mg/day. PMID- 8422190 TI - [Efficacy and safety of high dose NK 622 (toremifene citrate) in tamoxifen failed patients with breast cancer]. AB - Efficacy and safety of high dose administration of NK 622 (toremifene citrate) were studied in tamoxifen (TAM)--failed patients with breast cancer. The patients included in the study were the following failure cases in TAM therapy: unresponded cases in TAM therapy (TAM unresponded cases), temporary responded (CR, PR) but progressed cases in TAM therapy (TAM temporary responded cases), and those relapsing during TAM adjuvant therapy or within 6 months after the adjuvant therapy (TAM adjuvant failed cases). NK 622 of a 120 mg/day dose were orally given daily once at least for 8 weeks. The response rates in evaluable cases were 6.3% (1/16) in TAM unresponded cases, 11.1% (1/9) in TAM temporary responded cases, 15.4% (4/26) in TAM adjuvant failed cases, and 11.8% (6/51) in total cases including 1 CR and 5 PR cases. Long NC in which duration of NC maintained for more than 6 months was observed in 18.8% (3/16) of TAM unresponded cases, 22.2% (2/9) of TAM temporary responded cases, 11.5% (3/26) of TAM adjuvant failed cases, and 15.7% (8/51) of total cases. Rates of response and long NC were 14.3 and 19.0% in postmenopausal patients with estrogen receptor positive cancer, respectively. A median value of duration to the onset of response was 34 days (15 137). Median duration of response and long NC were 127 days (39-381) and 238.5 days (178-281), respectively. Adverse effects were experienced in 3 (5.1%) of 59 patients: nausea in 1, vertigo in 1 and increase of GOT, GPT, LDH and gamma-GTP in another 1. The side effects were moderate and reversible. From these results, NK 622 seems to become a safe and effective drug for TAM-failed patients with breast cancer by using a 120 mg/day dose. PMID- 8422191 TI - Low-dose fluconazole therapy potentiates the hypoprothrombinemic response of warfarin sodium. AB - BACKGROUND: Fluconazole has been reported to interact with many medications. This study examined the effect of low-dose fluconazole therapy on the hypoprothrombinemic response of warfarin sodium in patients. METHODS: Patients receiving low-intensity anticoagulation therapy with warfarin were recruited. All patients were taking stable doses of warfarin and had two baseline prothrombin times (PTs) within 10% of each other. Each patient received 100 mg of fluconazole daily for 7 days. Prothrombin times were measured on days 2, 5, and 8 during fluconazole administration. RESULTS: All patients had a progressive increase in PTs. Mean (+/- SD) of PTs increased from 15.8 +/- 1 seconds before the administration of fluconazole to 18.9 +/- 1.9 seconds on day 5 and 21.9 +/- 2.2 seconds on day 8. Fluconazole therapy was stopped early in the three patients due to high PTs. The largest change in PT was 9.7 seconds. No bleeding was noted during the study. CONCLUSION: Fluconazole predictably potentiates the hypoprothrombinemic response of warfarin. Prothrombin times must be monitored when fluconazole is administered to patients taking warfarin. PMID- 8422192 TI - Successful surgical management of neutropenic enterocolitis in two patients with severe aplastic anemia. Case reports and review of the literature. AB - We describe two patients with severe aplastic anemia in whom neutropenic enterocolitis developed while they were undergoing treatment at the National Institutes of Health. Both patients had progressive symptoms while receiving optimal medical management and both underwent and survived surgical intervention despite continued prolonged neutropenia in the perioperative period. This experience contrasts with six cases reported in the literature and suggests that surgery can be employed even in patients with profound neutropenia. Thus, in patients who remain persistently septic or who develop clinical deterioration despite medical management or who have other indications for surgical intervention, neutropenia should not be a contraindication to the appropriate or necessary procedure. PMID- 8422193 TI - Clinical and immunologic study of 205 patients with chronic fatigue syndrome: a case series from Italy. PMID- 8422194 TI - Quinolone resistance: a practical perspective. PMID- 8422196 TI - Nuts do not prevent heart attacks. PMID- 8422195 TI - Medical ethics--industry and realism. PMID- 8422197 TI - Left ventricular hypertrophy in diabetic patients. PMID- 8422198 TI - Freedom vs fairness. The need for a rational system of evaluation of medical trainees. PMID- 8422199 TI - Cigarette smoking, nicotine addiction, and its pharmacologic treatment. AB - Cigarette smoking is the most prevalent modifiable risk factor for increased morbidity and mortality in the United States and perhaps the world. Not only does the smoker incur medical risks attributable to smoking, passive smokers and society also bear ill effects and increased economic costs attributable to the smoker's habit. These risks of morbidity and mortality have been shown to be related to the addictive component of tobacco smoke, so that pharmacologic therapies have been studied in an attempt to modify the addiction, aid in smoking cessation, and prevent relapse. Presently, nicotine polacrilex and transdermal nicotine show some efficacy. Clonidine's efficacy has been equivocal. In addition, the combination of nicotine substitution and clonidine may be useful but will need formal investigation. Nicotine agonists and antagonists have not proven helpful. Antidepressants are being studied at this time. Nonpharmacologic modalities are briefly mentioned but play a major role in helping the smoker quit. PMID- 8422200 TI - Report of the Council on Scientific Affairs. Diet and cancer: where do matters stand? AB - During the past decade, the scientific literature base on the putative but elusive relationship between diet and cancer expanded enormously. Increased emphasis by funding agencies, fueled in turn by broadening public interest in the topic, led to this growth. The laboratory and epidemiologic research conducted in the past decade to determine the role of nutritional factors in the cause of cancer has shown that a simple solution does not exist. The key to the diet/cancer puzzle may lie in nutrient interactions and in individual response to dietary factors, determined in turn by genetic, physiologic, and life-style factors. Given the rapid strides being made in furthering the understanding of the biochemistry and molecular biology of cancer, it may be possible to look forward to the day when optimal dietary and life-style guidelines can be tailored on a specific individualized basis. PMID- 8422201 TI - Decreased risk of stroke among postmenopausal hormone users. Results from a national cohort. AB - OBJECTIVE: To assess the impact of postmenopausal hormone use on the risk of stroke incidence and stroke mortality. DESIGN: Longitudinal study consisting of three data collection waves. The average follow up for cohort members was 11.9 years (maximum, 16.3 years). Cox proportional hazards regression models were used to estimate the relative risk of stroke for postmenopausal hormone ever-users compared with never-users. PARTICIPANTS: A national sample of 1910 (of 2371 eligible) white postmenopausal women who were 55 to 74 years old when examined in 1971 through 1975 as part of the first National Health and Nutrition Examination Survey and who did not report a history of stroke at that time. MAIN OUTCOME MEASURE: The main outcome measure was incident stroke (fatal and nonfatal). Events were determined from discharge diagnosis information coded from hospital and nursing home records and cause of death information coded from death certificates collected during the follow-up period (1971 through 1987). RESULTS: There were 250 incident cases of stroke identified, including 64 deaths with stroke listed as the underlying cause. The age-adjusted incidence rate of stroke among postmenopausal hormone ever-users was 82 per 10,000 woman-years of follow up compared with 124 per 10,000 among never-users. Postmenopausal hormone use remained a protective factor against stroke incidence (relative risk, 0.69; 95% confidence interval, 0.47 to 1.00) and stroke mortality (relative risk, 0.37; 95% confidence interval, 0.14 to 0.92) after adjusting for the baseline risk factors of age, systolic blood pressure, diabetes, body mass index, smoking, history of hypertension and heart attack, and socioeconomic status. CONCLUSIONS: The results suggest that postmenopausal hormone use is associated with a decrease in risk of stroke incidence and mortality in white postmenopausal women. PMID- 8422202 TI - Metered-dose inhalers. Do health care providers know what to teach? AB - OBJECTIVE: The specific aim of this investigation was to evaluate the proficiency of health care providers and patients in the proper use of metered-dose inhalers. DESIGN, SETTING, AND PARTICIPANTS: Health care providers, which include house staff, nurses, and respiratory care practitioners who provide care to patients with asthma in the primary general medicine clinic or the pulmonary medicine clinic of a university-county hospital in which patients were referred, were surveyed and assigned a performance score regarding the knowledge base of the appropriate use of metered-dose inhalers. Patients who attended the primary care general medicine and pulmonary subspecialty clinic were also assessed as to their proficiency in the use of metered-dose inhalers. RESULTS: A significant percentage of patients had a poor understanding of the technique used with the metered-dose inhaler. House staff and nursing staff were also less proficient in the proper use of the metered-dose inhaler. The respiratory care practitioners were the most knowledgeable of the health care providers. CONCLUSIONS: This study confirms that a large percentage of patients use metered-dose inhalers improperly. It also demonstrates a significant lack of understanding by health care providers of the proper use of metered-dose inhalers. Furthermore, this study supports the use of respiratory care practitioners in the outpatient setting, since they were the most proficient among all the health care providers in the proper use of metered-dose inhalers. PMID- 8422203 TI - Do older Medicare patients cost hospitals more? Evidence from an academic medical center. AB - BACKGROUND: It is uncertain, under prospective payment, if hospitals face financial disincentives to treat older Medicare patients. Therefore, we studied associations between age and hospital charges and length of stay for Medicare patients and the impact on hospital reimbursement of Medicare's decision in October 1987 to eliminate older age (> or = 70 years) as a criterion for stratifying diagnosis-related groups (DRGs). METHODS: The 23,179 medical and surgical admissions to one academic medical center in 1985 through 1989 who were aged 65 years or more were studied using a retrospective cohort design. Clinical and financial data were obtained from hospital databases; charges and length of stay for each patient were adjusted for DRG weight, the measure used to determine reimbursement. Admission severity of illness was measured for 11,060 patients using the Nursing Severity Index, a previously validated method. RESULTS: Compared with patients aged 65 to 69 years, DRG-adjusted charges were 1%, 5%, 5%, and 6% higher and DRG-adjusted length of stay was 4%, 11%, 16%, and 18% greater for patients aged 70 to 74 years, 75 to 79 years, 80 to 84 years and 85 years or more, respectively. In multivariate analyses, these estimates were similar, even after controlling for sex, race, socioeconomic status, and other variables associated with charges and length of stay. However, further controlling for severity of illness revealed that nearly all of the differences in charges and a large proportion of the differences in length of stay in older patients could be explained by their higher severity of illness. In separate stratified analyses, the association with age was stronger and more consistent in patients admitted after October 1987 and in medical patients. CONCLUSIONS: These findings suggest that currently hospitals may face financial disincentives to care for older Medicare patients and that the equitability of DRG-based hospital payments, with respect to age, may have been adversely affected by Medicare's decision to eliminate older age (> or = 70 years) as a criterion for classifying DRGs. The inclusion of patient age in prospective payment formulas may make hospital reimbursement more equitable. PMID- 8422204 TI - Resistance exercise training is associated with decreases in serum low-density lipoprotein cholesterol levels in premenopausal women. AB - BACKGROUND: Aerobic exercise training is associated with reduced serum concentrations of triglycerides, increased concentrations of high-density lipoprotein cholesterol, and minimal changes in serum levels of total cholesterol or low-density lipoprotein cholesterol. There are few data on the effects of resistance exercise on blood lipid levels. METHODS: Premenopausal women were randomly assigned to a supervised resistance exercise training program (n = 46) or a control group (n = 42) for 5 months. Serum was analyzed for levels of total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglycerides. Body composition and dietary intake were also measured. RESULTS: The exercise group showed a 0.33 +/- 0.03-mmol/L (mean +/- SE) decrease in total cholesterol level and a 0.36 +/- 0.001-mmol/L decrease in low density lipoprotein cholesterol level that was significantly different from the control group. No significant changes were noted in serum high-density lipoprotein cholesterol or triglyceride levels in either group. Changes in body composition showed no significant correlations with changes in total cholesterol or low-density lipoprotein cholesterol. There were no significant differences in nutrient intake between the groups. CONCLUSION: In healthy, premenopausal women, with normal baseline lipid profiles, 5 months of resistance exercise training was associated with significant decreases in serum total cholesterol and low-density lipoprotein cholesterol concentrations. PMID- 8422205 TI - Hypertension: steps forward and steps backward. The Joint National Committee fifth report. PMID- 8422206 TI - The fifth report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure (JNC V) PMID- 8422207 TI - National High Blood Pressure Education Program Working Group report on primary prevention of hypertension. PMID- 8422208 TI - Tuberculosis infection among young adults entering the US Navy in 1990. AB - BACKGROUND: From 1958 through 1969, more than 1.2 million US Navy recruits received tuberculin skin tests; 5.2 per 100 were tuberculin reactors. Subsequent analyses predicted a downward trend in the risk of tuberculosis infection in the United States. We sought to determine the current prevalence of tuberculin reactors by sex, race/ethnic group, and birthplace among young adult residents of the United States entering the US Navy. METHODS: Recruits routinely receive a tuberculin skin test on entering US Navy recruit training in Great Lakes, Ill, Orlando, Fla, or San Diego, Calif. In January and February 1990, 2416 young men and women (mean age, 20.6 years) received tuberculin skin tests and completed questionnaires eliciting demographic and tuberculosis risk factor data. A tuberculin reactor was defined as a subject having 10 mm or greater induration to a skin test with 5 tuberculin units, purified protein derivative, administered intradermally by the Mantoux method. RESULTS: Fifty-five of 2214 men (2.5 per 100; 95% confidence interval, 1.9 to 3.2 per 100) and five of 202 women (2.5 per 100; 95% confidence interval, 0.8 to 5.8 per 100) were tuberculin reactors. For men, the prevalence was greater in blacks (5.2 per 100), Hispanics (5.4 per 100), and Asian/Pacific Islanders (26.4 per 100) than in whites (0.8 per 100) and greater in foreign-born recruits (19.2 per 100) than in recruits born in the United States (1.6 per 100). Women had the same pattern of prevalence by race/ethnic group and birthplace. CONCLUSIONS: The prevalence of tuberculosis reactors declined as predicted among young adults, especially the white US-born recruits, entering the US Navy. Although the prevalence also declined among nonwhites and the foreign-born recruits, a substantial proportion continue to enter adulthood with preexisting tuberculosis infection. PMID- 8422209 TI - Cost of care for patients with human immunodeficiency virus infection. Patterns of utilization and charges in a public health care system. AB - BACKGROUND: The increasing impact of human immunodeficiency virus (HIV) infection on the health care delivery system requires surveillance of current patterns of HIV-related health care utilization to adequately plan for future needs. Most studies to date have concentrated on inpatient care for patients with the acquired immunodeficiency syndrome (AIDS). Outpatient utilization has been less well studied and there are few data regarding HIV-infected patients without a diagnosis of AIDS. METHODS: Denver Health and Hospitals is a public system delivering comprehensive health care to mostly indigent residents of the city and county of Denver. Patients with HIV infection in this system were identified through multiple surveillance sources, and billing system records for these patients were analyzed. RESULTS: During 1990, 812 patients with HIV infection of 13 years or more were accessed in the Denver Health and Hospitals. During that year, the total HIV-related health care charges were $7,858,690, of which 57% were for inpatient care and 43% for ambulatory care. Patients with AIDS (34% of patients) accounted for 62% of all charges, and patients with HIV infection but without a diagnosis of AIDS (66% of patients) for 38% of charges. Compared with national predictions, patients with AIDS in our system had lower inpatient and higher outpatient utilization. CONCLUSIONS: These results are consistent with a shift from inpatient to outpatient health care services in patients with AIDS. A significant proportion of HIV-related health care costs are incurred by patients who have not yet developed AIDS. PMID- 8422210 TI - Impact of a procedure-specific do not resuscitate order form on documentation of do not resuscitate orders. AB - BACKGROUND: Serious problems exist with respect to documentation of do not resuscitate (DNR) orders. We studied the impact of a procedure-specific DNR order form on documentation of these orders. METHODS: We prospectively compared DNR chart documentation during a 3-month period before and after implementation of a procedure-specific DNR order form. RESULTS: The order form was used in 41 (93%) of 43 charts after its implementation. Documentation of attending physician agreement with the DNR order form increased from 30 of 34 charts in which the order form was used). The number of orders where it was uncertain whether at least one component of acute cardiopulmonary life support-related procedures was to be performed decreased from 30 (88%) of 34 charts to three (7%) of 43 charts. The order form had no measurable impact on documentation of DNR discussion. Only 25% of the charts had any discussion of the risks and benefits of CPR. CONCLUSIONS: A procedure-specific DNR order form can improve documentation of DNR decisions. The reduction of uncertainty in these orders about the use of specific procedures can prevent errors in patient care. PMID- 8422211 TI - Prevalence and determinants of acute renal failure following cardiopulmonary resuscitation. AB - BACKGROUND: The purpose of this study was to determine the prevalence and determinants of acute renal failure in patients following cardiac arrest. METHODS: This was a cross-sectional study of 420 consecutive admissions with a diagnosis of cardiac arrest admitted to the Long Island Jewish Medical Center, New Hyde Park, NY, the Long Island Campus for the Albert Einstein College of Medicine, Bronx, NY, over a 2-year period. Fifty-six patients who initially survived cardiopulmonary resuscitation following cardiac arrest and had serial biochemical and renal function data available were studied. The events during cardiopulmonary resuscitation and clinical and biochemical data were compared and contrasted among patients who developed acute renal failure following cardiopulmonary resuscitation (group 1, n = 16) and those who did not (group 2, n = 40). RESULTS: Patients who developed acute renal failure following cardiopulmonary resuscitation (group 1) had longer duration of resuscitation (12.0 +/- 2.1 minutes vs 6.7 +/- 0.9 minutes for group 2) and received larger dosages of epinephrine during cardiopulmonary resuscitation (1.81 +/- 0.36 mg vs 0.90 +/- 0.18 mg for group 2). Patients in group 1 had a significantly higher frequency of congestive heart failure (43.8% vs 12.5% for group 2), coronary artery disease (87.5% vs 37.5% for group 2), and preexisting compromised renal function (50% vs 12.5% for group 2). Patients in group 1 had significantly worsened long-term survival compared with group 2 patients (6.3% vs 47.5% for group 2). CONCLUSIONS: We conclude that acute renal failure occurs commonly in the postcardiac arrest period. Administration of the vasoconstrictor epinephrine, congestive heart failure, coronary artery disease, and preexisting renal insufficiency may be significant risk factors for the development of postcardiac arrest acute renal failure. The development of acute renal failure following cardiopulmonary resuscitation predicts a lesser likelihood of survival to discharge from the hospital. PMID- 8422212 TI - Reporting of age data in clinical trials of arthritis. Deficiencies and solutions. AB - OBJECTIVE: We studied recent nonsteroidal anti-inflammatory drug (NSAID) randomized control trials of arthritis to identify the age and number of older people (> or = 65 years) and to document the way information on age was presented. We hypothesized that older people, who are most likely to take NSAIDs are underrepresented and underreported. STUDY SELECTION AND DATA EXTRACTION: All NSAID articles (n = 1008) in MEDLINE between September 1987 and May 1990 were identified. Eighty-three trials employing NSAIDs in a randomized control trial of arthritis reported in 73 articles were identified and studied in detail for age related information. RESULTS: A total of 9664 subjects with a female-to-male ratio of 2.3:1 were enrolled. Forty-four trials studied osteoarthritis (53.0%), 37 studied rheumatoid arthritis (44.6%), and two studied both conditions (2.4%). More than half of the studies reviewed included people 65 years of age or older, only 207 people in this older age group could be identified (2.1%). While there was inclusion of the 'young-old' (65 to 74 years of age), only 14 of the 9664 people studied were between 75 and 84 years of age, and no one 85 years or older could be identified. The inclusion of the young-old is documented by the weighted mean age that ranged from 59.6 to 64.9 years for patients with osteoarthritis (mean, 62.9; SD, 1.67) and from 47.4 to 53.0 years (mean, 49.9; SD, 2.16) for those with rheumatoid arthritis. CONCLUSION: We demonstrate that older people, who represent a high proportion of the population treated with NSAIDs in practice, are generally omitted from drug trials. Recommendations designed to improve the reporting of age information to make clinical trials more informative and applicable to older people are presented. PMID- 8422213 TI - Spontaneous remission in Cushing's disease. AB - Two patients with Cushing's disease underwent spontaneous clinical and biochemical remission following a period of secondary adrenal insufficiency. One of the patients was untreated, while the other had been treated with partial bilateral adrenalectomy before. Both patients underwent remission shortly after the administration of dexamethasone for diagnostic purposes and remained symptom free for 2 and 3 years, respectively, which was followed by relapse of the disease. We conclude that although resolution of Cushing's disease may occur spontaneously, probably by hemorrhagic infarction of corticotroph microadenomas, careful follow-up studies are required since the relapse of the disease occurs later in some patients. PMID- 8422214 TI - Dichotomy in health care policy. PMID- 8422215 TI - How was 'race' determined? PMID- 8422216 TI - Site for subcutaneous heparin injection. PMID- 8422217 TI - Seasonal mood disorders. Patterns of seasonal recurrence in mania and depression. AB - DSM-III-R criteria, applied retrospectively in a research-oriented psychiatric clinic, identified patients (N = 146) with a mood disorder and a seasonal pattern of recurrence (seasonal mood disorder). The seasonal mood disorder syndrome was not rare (10% of all mood disorders); diagnostic distribution was as follows: recurrent depression, 51%, and bipolar disorder, 49%, with 30% of the latter having mania (bipolar disorder type I) and 19% having hypomania (bipolar disorder type II). Most patients were women (71%); onset age averaged 29 years, with a mean of eight cycles in 12 years of illness; mean episode duration was 5.0 months. Mood disorder was found in a high proportion (68%) of the families. All but one patient followed one of two seasonal patterns in equal frequency: type A, fall-winter depression with or without spring-summer mania or hypomania; and type B, spring-summer depression with or without fall-winter mania or hypomania. Both types showed consistent times of onset and remission. These results emphasize that DSM-III-R seasonal mood disorder includes severe cases of recurrent depression and bipolar disorder and support a distinction between two seasonal subtypes. PMID- 8422218 TI - Sexual function in depressed men. Assessment by self-report, behavioral, and nocturnal penile tumescence measures before and after treatment with cognitive behavior therapy. AB - Clinicians have long associated depression with alterations in sexual function, predominantly loss of sexual interest. In a longitudinal study measuring self report, behavioral, and nocturnal penile tumescence variables before and after treatment with cognitive behavior therapy in an unmedicated sample of 40 outpatient depressed men, we found, contrary to expectation, that sexual activity per se was not reduced during the depressed state. Rather, loss of sexual interest appeared to be related to the cognitive set of depression, ie, loss of sexual satisfaction that then improved with remission from depression. Depressed men were heterogeneous, however, with respect to sexual behavior, eg, an anxious and more chronically depressed subgroup of men who did not have remissions with cognitive behavior therapy reported increased sexual interest and sexual activity. Also, contrary to expectation, nocturnal penile tumescence abnormalities in depressed men did not reverse when measured in early remission, nor did nocturnal penile tumescence measures correlate significantly with behavioral measures of sexual function. Nocturnal penile tumescence alterations in depression may thus be similar to other persistent electroencephalographic sleep abnormalities seen in depressed patients in remission, in being more trait like than statelike. PMID- 8422219 TI - Nicotine dependence and major depression. New evidence from a prospective investigation. AB - We examined prospectively the association between nicotine dependence and major depression (MDD). The following questions were addressed: (1) Are smokers with a history of MDD at increased risk for progression to nicotine dependence and more severe levels of dependence? (2) Are persons with a history of nicotine dependence at increased risk for MDD? A sample of 995 young adults were interviewed in 1989 and reinterviewed in 1990, 14 months later. The revised National Institute of Mental Health-Diagnostic Interview Schedule was used to ascertain DSM-III-R nicotine dependence and other substance use and psychiatric disorders. A history of MDD increased the risk for progression to nicotine dependence or more severe levels of dependence (odds ratio, 2.06; 95% confidence interval, 1.21 to 3.49). In addition, persons with a history of nicotine dependence had a higher rate of first-incidence MDD during the follow-up period than persons with no history of nicotine dependence (7.5% vs 3.2%; odds ratio, 2.45; 95% confidence interval, 1.17 to 5.15). The prospective data suggest that the association between nicotine dependence and MDD, observed previously in cross sectional studies, might be either causal, with influences flowing in both directions, or, more probably, noncausal, reflecting the effects of common factors that predispose to both disorders. PMID- 8422220 TI - Smoking and major depression. A causal analysis. AB - Among 1566 personally evaluated female twins from a population-based register, average lifetime daily cigarette consumption was strongly related to lifetime prevalence and to prospectively assessed 1-year prevalence of major depression (MD). Using the cotwin control method, we evaluated whether the association between smoking and lifetime MD was causal or noncausal. While the relative risk (95% confidence interval) for ever smoking given a lifetime history of MD was 1.48 (1.30 to 1.65) in the entire sample, it was 1.18 (0.88 to 1.47) and 0.98 (0.71 to 1.26), respectively, in dizygotic and monozygotic twin pairs discordant for a history of MD. The relative risk for a history of MD given ever smoking was 1.60 (1.39 to 1.83) in the entire sample, while in dizygotic and monozygotic twins discordant for smoking, it was 1.29 (0.87 to 1.74) and 0.96 (0.59 to 1.42), respectively. Controlling for personal smoking history, family history of smoking predicted risk for MD; controlling for the personal history of MD, family history of MD predicted smoking. The best-fitting bivariate twin model suggested that the relationship between lifetime smoking and lifetime MD resulted solely from genes that predispose to both conditions. These results suggest that the association between smoking and MD in women is not a causal one but arises largely from familial factors, which are probably genetic, that predispose to both smoking and MD. PMID- 8422221 TI - A comparison of fluvoxamine, cognitive therapy, and placebo in the treatment of panic disorder. AB - Seventy-five outpatients with moderate to severe panic disorder were randomly assigned to receive 8 weeks of fluvoxamine, cognitive therapy, or placebo. Fifty five patients completed the treatment protocol. Fluvoxamine was found to be an effective and well-tolerated treatment for panic using clinician- and patient rated variables. Subjects receiving cognitive therapy also showed improvement, but this improvement did not significantly differ from the experience of the placebo-treated group for most comparisons. Fluvoxamine was superior to cognitive therapy for many ratings, but cognitive therapy was not superior to fluvoxamine on any rating. Fluvoxamine also produced improvement earlier than cognitive therapy. At the main comparison point (week 4), 57% (13/23) of patients receiving fluvoxamine were rated moderately improved or better vs 40% (8/20) of the group given cognitive therapy and 22% (5/23) of the placebo-treated group. At that point, 43% (10/23) of the fluvoxamine recipients vs 25% (5/20) of cognitive therapy and 4% (1/23) of placebo recipients were free of panic attacks. PMID- 8422222 TI - Maintenance drug treatment of panic disorder. I. Results of a prospective, placebo-controlled comparison of alprazolam and imipramine. AB - One hundred six patients diagnosed according to DSM-III as suffering from agoraphobia with panic disorder, panic disorder with limited phobic avoidance, or uncomplicated panic disorder entered an acute 8-week treatment phase. Patients who improved received an additional 6 months' maintenance treatment. Significantly more patients treated with alprazolam than with imipramine hydrochloride or placebo remained in therapy and experienced panic attack and phobia relief during the acute treatment phase. During the maintenance phase, neither tolerance nor daily dose increase was observed. All patients who completed the maintenance phase (27 in the alprazolam group, 11 in the imipramine group, and 10 in the placebo group) were panic free at the end of 8 months of study treatment. Alprazolam therapy was effective and well tolerated at a mean daily dose of 5.7 mg. Imipramine hydrochloride (175 mg/d) also produced significant panic relief but was associated with poor patient acceptance. PMID- 8422223 TI - Maintenance drug treatment for panic disorder. II. Short- and long-term outcome after drug taper. AB - Forty-eight patients with panic disorder completing 8 months of maintenance treatment with alprazolam (mean dose, 5.2 mg [n = 27]), imipramine hydrochloride (mean dose, 175 mg [n = 11]), or placebo (mean dose, 8.0 pills [n = 10]) underwent a gradual taper from medication over a 4-week period. A withdrawal syndrome was observed in almost all alprazolam-treated patients but in only a few imipramine- or placebo-treated patients. The clinical worsening of withdrawal symptoms after discontinuation tended to subside over the course of 3 medication free weeks, but 33% of alprazolam-treated patients were unable to discontinue their medication regimen successfully. Severity of panic attacks at baseline but not daily alprazolam dose appeared as a significant independent predictor of taper difficulty. Forty-nine percent of the total study population continue to receive drug therapy: 82% alprazolam and 18% imipramine. Among patients who received alprazolam during study treatment and at follow-up, the mean daily dose was substantially reduced (6.1 vs 1.6 mg [n = 14]). At follow-up, after 1 year of naturalistic treatment for panic symptoms and combining 8-month completers and study dropouts, there were no significant differences in remission (68% to 71%) or in antipanic medication intake (39% to 56%) at follow-up for the three original treatment groups. However, 8-month study completers compared with study dropouts had a significantly higher remission rate (85% vs 55%). PMID- 8422224 TI - Quantitative cerebral anatomy in depression. A controlled magnetic resonance imaging study. AB - Magnetic resonance imaging was used to examine cerebral anatomy in 48 inpatients with severe depression who were referred for electroconvulsive therapy and in 76 normal control subjects. The magnetic resonance imaging measures included determinations of regional cerebral volumes and ratings of the frequency and severity of cortical atrophy, lateral ventricular enlargement, and subcortical hyperintensity. The mean total frontal lobe volume was found to be 7% smaller in the inpatients with severe depression (235.88 mL) than in the normal control subjects (254.32 mL)--a difference that was statistically significant even after adjusting for the effects of age, sex, education, and intracranial size. No group differences were observed in the volumes of the cerebral hemispheres, the temporal lobes, or the amygdala-hippocampal complex, nor in the frequency of cortical atrophy. Neither did the groups differ with respect to the total volumes of the lateral and third ventricles, nor in the frequency of lateral ventricular enlargement. Patients with depression had a significantly higher frequency of subcortical hyperintensity in the periventricular white matter, with an odds ratio of 5.32. PMID- 8422225 TI - What is the 'core' symptom of mania? PMID- 8422226 TI - Lithium treatment increases norepinephrine turnover in the plasma of healthy subjects. PMID- 8422227 TI - A balanced 2:18 translocation and familial schizophrenia: falling short of an association. PMID- 8422228 TI - Dependence of baroreceptor-mediated sympathetic outflow on biventricular assist device driving frequency. AB - To investigate the drive condition of the artificial heart from a neurophysiological point of view, the dynamic transduction characteristics of the baroreflex system were analyzed by means of sympathetic neurograms. Two pneumatically actuated ventricular assist devices were implanted as biventricular bypasses (BVBs) in adult mongrel dogs to compare the natural heart circulation and prosthetic circulation. After BVB pumping was initiated, the natural heart was electrically fibrillated. Renal sympathetic nerve activity (RSNA) was recorded to analyze sympathetic outflow. Coherence function between the arterial pressure and RSNA was calculated to evaluate the linearity of the baroreflex system. The largest peak coherency was observed when BVB was driven at the frequency of natural heart beat prior to electrical fibrillation, which suggests that the baroreflex system shows the largest linearity at this frequency. These results suggest the possibility that the natural heart beat frequency is the setting frequency at which the baroreflex system transfers the hemodynamic rhythm to the sympathetic outflow. PMID- 8422229 TI - Analysis of phosphate kinetics in hemofiltration and hemodiafiltration: formulation of an efficiency index. AB - The kinetics of phosphate (P) during a treatment of hemofiltration or hemodiafiltration follows a simple exponential law, and P in 90 min reaches a stationary level that remains constant until the end of the treatment. The theoretical compartmental model cannot be estimated by a dialytic treatment, but a simpler diffusive model may be considered. Once the stationary level is reached, the net flux of P from the inaccessible compartments to the accessible compartment may be estimated via the parameters obtained in the exponential model. Hemodiafiltration seems more efficient than hemofiltration in determining P clearance. Thus an efficiency index that accounts for the period in which P is eliminated from the accessible compartment has been formulated. PMID- 8422230 TI - Improved liver function following treatment with an extracorporeal liver assist device. AB - A 68-year-old woman with fulminant hepatic failure of unknown etiology was treated with a bioartificial liver assist device. Prior to treatment, she had a number of clinical and laboratory features that suggested a hopeless outcome. Treatment was associated with a dramatic change in her mental status, and her clinical picture improved progressively during 6 days of continuous therapy. Evidence of recovery of native liver function allowed the discontinuation of treatment, and she continued to improve for a further 3 1/2 days before her demise from septic shock. We propose that a metabolically active liver assist device is a logical and practical method for treating the critical phase of fulminant hepatic failure. PMID- 8422231 TI - Alternative descriptions of combined diffusive and convective mass transport in hemodialyzer. AB - Two alternative versions of the mathematical description of the combined diffusive and convective membrane transport in hemodialyzers were compared using the one-dimensional theory of hemodialyzers. The first version is based on the assumption of homogeneity of the membrane. The second version is a widely used "ad hoc" formulation, which can be interpreted as a description of the membrane as tighter at the dialysate side than at the blood side. Theoretical predictions of the increase of dialyzer clearance caused by ultrafiltration, as assessed by transmittance coefficient, were compared to experimental data about transport of small solutes (urea, creatinine, and sodium) as well as middle molecules (vitamin B12) in three types of hollow-fiber hemodialyzer. For one type of dialyzer, the theory assuming the homogeneous membrane yielded the correct predictions for the small solutes. For two other types of dialyzer, the alternative version of the theory was adequate. For vitamin B12, the experimental values of transmittance coefficient were between the values predicted by the two versions of the theory for all three types of hemodialyzer. Thus, the two versions should be considered as a possible adequate description of solute transport in hemodialyzers. PMID- 8422232 TI - Experimental results using nafamostat mesilate as anticoagulant during extracorporeal lung assist for 24 hours in dogs. AB - We report on the experimental application of nafamostat mesilate (NM, 6-amidino-2 naphthyl p-guanidinobenzoate dimethanesulfonate, FUT), a new anticoagulant, to extracorporeal lung assist (ECLA) with an artificial membrane lung. Venovenous ECLA, from the jugular vein to the femoral vein, was performed with a hollow fiber membrane lung at a blood flow rate of approximate 82 ml kg-1 min-1 for 24 h in 7 dogs under anesthesia and hypoventilation. Heparin (10 U ml-1 in a priming lactated Ringer solution of 140 ml, and 200 U kg-1) was administered before blood access cannulation. After start of ECLA, however, no heparin was used, and nafamostat mesilate was continuously infused into the drainage line of the bypass circuit to control activated coagulation time (ACT) at about 150 to 200 s. To maintain the prolonged ACT, 8.0 +/- 1.7 mg kg-1 h-1 of NM was required. Arterial blood pressure and pulse rate decreased significantly. Though fibrin monomer test revealed hypercoagulability after 6 h of ECLA, platelet counts did not significantly decrease. Total blood loss remained less than 40 g. The artificial membrane lung sustained a good gas exchange and low flow resistance throughout ECLA. Macroscopic examination revealed small spotty thrombi in the artificial lung but no major pathologic changes of the visceral organs in the all dogs at autopsy. High-dose NM administration could control blood coagulation and decrease blood loss during ECLA for 24 h without deterioration of the artificial lung and systemic complication other than mild hypotension and bradycardia. PMID- 8422233 TI - A seal-less centrifugal pump (Baylor Gyro Pump) for application to long-term circulatory support. AB - We are developing a new centrifugal pump, the Baylor Gyro Centrifugal Pump (Gyro Pump), which can function for more than 2 weeks. The concept of the Gyro Pump is that a one-piece rotor-impeller with embedded permanent magnets, driven directly by a brushless direct current motor stator placed outside, rotates like a "gyroscope," and the rotor-impeller is supported by one pivot bearing at the bottom in accordance with the gyroscopic principle. This concept enables us to eliminate a driving shaft and a seal between the driving shaft and the blood chamber, which results in extending the life of the centrifugal pump. The blood passes through the space between the motor stator and the rotor to the impeller portion. In this preliminary phase, two pivot bearings were applied to support the rotor-impeller at the top and the bottom inside the blood chamber. Both pivot bearings showed less blood trauma and less thrombogenicity in in vitro and in vivo studies. The Gyro Pump is a promising second-generation centrifugal pump for long-term circulatory support in the near future. PMID- 8422234 TI - Liver assist devices (LADs) will not be used to treat fulminant hepatic failure (FHF), but its consequences, namely hepatic encephalopathy (HE) PMID- 8422235 TI - Iontophoresis of drugs in the bladder wall: equipment and preliminary studies. AB - Iontophoresis is the active transport of ions into tissues by means of an electric current: Ji = -D(i)delta Ci/delta chi + DizeECi/kT. Where Ji is the total ionic flux, D(i) the diffusion coefficient, Ci the concentration, z the valency, and E the electric field. The first expression on the right side of the equation is Fick's law of diffusion and approaches zero for bladder mucosa, which leads to uncertain results following intravesical administration of various therapeutic agents. The application of an electric field will potentially accelerate drug administration into the bladder wall in a controllable manner. To evaluate this concept, an appropriate source of electric current and electrodes was fabricated; then, studies were conducted in human cadaveric bladders and clinical trials in human subjects. Ionized dyes were applied in duplicate to 10 fresh cadaveric bladders. Electric currents (3.5-5.0 mA) were applied for 20 min to 10 solutions, and no current was used in 10 controls. Twenty-eight patients had 100 ml solutions of 1% mepivacaine or lidocaine with epinephrine infused into their bladders prior to endoscopic resections. Twenty-two patients received currents of 10-20 mA for 10-20 min, and 6 controls had either no drugs or a current of reverse polarity applied. Visually and on microscopy, the 10 control cadaveric bladder surfaces demonstrated only faint staining of the surface mucosa whereas the experimental surfaces showed full-thickness staining of the mucosa extending into the muscularis. The 6 control patients required supplemental anesthesia or abandonment of the operative procedure. Of the 22 experimental subjects, 16 tolerated procedures with up to 25 g of tissue removed by diathermic resection. PMID- 8422236 TI - Cloning of a functional cDNA for human cytidine deaminase (CDD) and its use as a marker of monocyte/macrophage differentiation. AB - We have identified a cDNA clone for human cytidine deaminase (EC 3.5.4.5) during an investigation which aimed at cloning novel gene expression products related to monocyte/macrophage differentiation. The derived amino acid sequence of the clone comprises 145 residues yielding a molecular mass for the polypeptide of 16.1 kDa and exhibits a nearly 50% homology to cytidine deaminase from Bacillus subtilis. Cytidine deaminase activity of the cloned sequence could be demonstrated in a prokaryotic expression system. The mRNA is highly expressed in granulocytes while expression is very low in fibroblasts, chondrocytes, monocytes, and T- as well as B-cell lines. The mRNA can be induced in monocytes, the monocytoid cell line U937 and the myeloblastic line HL 60 by the differentiation inducer calcitriol. PMID- 8422237 TI - Rat CINC (cytokine-induced neutrophil chemoattractant) is the homolog of the human GRO proteins but is encoded by a single gene. AB - Human GRO and rat CINC proteins are members of the pro-inflammatory "chemokine" superfamily of chemotactic cytokines which includes IL-8. We have used Northern and Southern blot analyses to compare the expression of CINC-related and GRO transcripts in human cells and to compare the hybridization patterns of CINC and GRO probes in human and rat genomic DNA. Our data indicate that rat CINC is encoded by a single gene unlike GRO-alpha, -beta and -gamma which are encoded by three distinct genes and that they are the nearest homologs to each other from their respective species. PMID- 8422238 TI - Beta 3 integrin derived peptide 217-230 inhibits fibrinogen binding and platelet aggregation: significance of RGD sequences and fibrinogen A alpha-chain. AB - beta 3 Integrin derived peptides 217-230 (DAPEGGFDAIMQAT) and 217-231 (Y) (DAPEGGFDAIMQATVY) at 100 microM inhibited 125I-fibrinogen binding to ADP stimulated platelets and platelet aggregation. Peptide 217-231 (Y) (100 microM) significantly inhibited the binding of 125I-albolabrin (a disintegrin with a single RGD sequence) to ADP- and thrombin-activated platelets while it had only a slight effect on albolabrin binding to resting platelets. The 125I-beta 3 217-231 (Y) cross-linked selectively to the fibrinogen A alpha-chain. The interaction of the RGD sequence in the A alpha-chain of fibrinogen with beta 3 217-231 sequence appears to play a significant role in the events leading to platelet aggregation. PMID- 8422239 TI - Expression of an N-terminally truncated form of human focal adhesion kinase in brain. AB - We have cloned a novel tyrosine kinase that is widely expressed in human tissues using degenerate oligonucleotide primers. The cDNA clone was subsequently found to be the human homologue of the recently cloned chicken focal adhesion associated kinase (pp125FAK). The homology between the chicken and human sequences is 95% at the amino acid level. By RT-PCR we have detected hFAK in human tonsillar T and B cells, several human lymphoid cell lines, a neuroblastoma cell line and HeLa cells. By Northern blot analysis we show that hFAK is expressed in all organs tested with the highest abundance in brain and the least in heart and skeletal muscle. An additional transcript of ca. 3.3 kb, encoding an N-terminally truncated form of hFAK, was observed only in brain. PMID- 8422240 TI - Induction of apoptosis by hemorrhagic snake venom in vascular endothelial cells. AB - Vascular degeneration appears to play crucial roles in producing many vascular malfunctions (1-3). In order to identify specific inducers of programmed death in vascular endothelial cells (VEC), examinations were made of the effects of substances that are known to affect the vascular system by using VEC in culture (4,5). We found that hemorrhagic snake venoms induced apoptotic cell death or programmed cell death of VEC. By contrast, neurotoxic snake venoms did not induce programmed cell death but caused necrosis at much higher doses of the venoms. No effect of hemorrhagic venom was observed with many types of cultured cells other than VEC. Thus, hemorrhagic snake venom appears to be a useful tool for studies of the molecular mechanisms of vascular apoptosis. The results also suggest a possible mechanism of action of hemorrhagic snake venom on the vascular system. PMID- 8422241 TI - Interleukin 1 receptor on fibroblasts from systemic sclerosis patients induces excessive functional responses to interleukin 1 beta. AB - To determine if SSc fibroblasts are more sensitive to exogenous IL-1 than are normal fibroblasts, we studied the inductions of IL-6, prostaglandin E2 (PGE2), IL-1 beta mRNA production by IL-1-stimulated SSc and normal fibroblasts. The minimal IL-1 beta concentrations for stimulation of IL-6 and PGE2 production by SSc fibroblasts were 10-fold and 100-fold lower, respectively, than those for normal fibroblasts. The minimal IL-1 beta concentration for stimulation of IL-1 beta mRNA expression by SSc fibroblasts was also 10-fold lower than that for normal fibroblasts. These results suggest that the IL-1 signal transduction through IL-1R could be induced excessively in SSc fibroblasts. It is suggested that the IL-1 signal transduction plays an important role in the abnormal cytokine networks observed in the fibrosis of SSc. PMID- 8422242 TI - Cloning of a cDNA encoding rat intestinal 15 kDa protein and its tissue distribution. AB - A cDNA encoding rat intestinal 15 kDa protein was isolated and sequenced from a rat ileum-specific cDNA library. This cDNA was found to contain an open reading frame of 384 nucleotides as well as 5' (27 nucleotides) and 3' (46 nucleotides) non-coding regions. The deduced sequence of 127 amino acids was identical to that of rat I-15P which was purified from rat intestinal epithelium. The nucleotide sequence of the open reading frame exhibited 79% identity to that of the porcine gastrotropin. Northern blot analysis indicated that the same size of transcript as that of the ileum was detected in the ovary, suggesting that I-15P or a homologous protein might be involved in the metabolism of steroids in steroid hormone-producing tissues. PMID- 8422243 TI - Nuclear translocation of aflatoxin B1 - protein complex. AB - The in vitro binding of [3H]-AFB1 to various proteins was studied by equilibrium dialysis. At 23 +/- 1 degree C, [3H]-AFB1 binding activity (mmol/mol) decreased as follows: pyruvate kinase > albumin-NLS > albumin > carbonic anhydrase > RNase > histones. The nuclear translocation and activation of AFB1 and AFB-protein complexes was investigated using isolated rat liver nuclei in the presence of ATP and a NADPH regenerating system. Proteins containing NLS such as histones and albumin-NLS facilitated AFB1 translocation into the nucleus where activation and adduct formation took place. PMID- 8422244 TI - The disulfide bond arrangement of leukemia inhibitory factor: homology to oncostatin M and structural implications. AB - Murine leukemia inhibitory factor (LIF) (the fully active recombinant form produced in E. coli) was digested in the unreduced state with trypsin and Staphylococcal V8 protease in 0.05% sodium dodecyl sulfate. Disulfide-bonded peptides were identified by altered mobility on reverse-phase high-performance liquid chromatography in the presence or absence of dithiothreitol and subjected to amino acid sequencing. Peptides containing more than one disulfide bond were subjected to further proteolysis and disulfide-bonded subfragments identified and sequenced. The three disulfide bonds are CYS13-CYS135, CYS19-CYS132 and CYS61-164 and the first and third of these are clearly homologous to the two disulfide bonds in oncostatin M. The spatial organization of the cysteine residues in the predicted four alpha-helical bundle structure of LIF (Bazan, Neuron 7,197;1991) is compatible with these disulfide assignments. PMID- 8422245 TI - Quinone analogues: a drug of choice for the control of filariasis. AB - Human filariasis is caused by Wuchereria bancrofti, Brugia malayi and B. timori. Of the several recommended model filarial parasites by WHO, Setaria digitata a bovine one has characteristics such as cyanide insensitivity, lack of detectable cytochromes, presence of two quinones Q8 and Q6. Of the two quinones Q8 seems to have a predominant role in energy production. In vitro inhibitory studies using quinone analogues, coenzyme Q0 and menadione have shown that these compounds paralyse the worms in very low concentrations compared to diethyl carbamazine, the drug of choice for filariasis. The mitochondrial energy production associated with electron transfer is intercepted by quinone analogues. Hence for the treatment of filariasis, this study paves a chemotherapeutic target for the design of drugs which can control the parasites by interacting at the subcellular level by energy depletion. PMID- 8422246 TI - A proteoglycan form of macrophage colony-stimulating factor that binds to bone derived collagens and can be extracted from bone matrix. AB - We previously found that the human osteoblastic cell line MG-63 produces two molecular types of macrophage colony-stimulating factor (M-CSF). One is an 85-kD M-CSF, and the other is a proteoglycan form of M-CSF (PG-M-CSF) that has a binding affinity to type V collagen. The latter type of M-CSF showed dose dependent binding to wells coated by pepsin-extracted bone collagens, whereas the 85-kD M-CSF did not. Immunoblot analysis of urea-extracted bone M-CSF revealed the presence of PG-M-CSF. PG-M-CSF contained in bone matrix may have physiological importance in the bone metabolism. PMID- 8422247 TI - Modulation of guanine triphosphate nucleotide binding to p21ras immunoprecipitates of rat liver plasma membranes by agents affecting redox state. AB - GTP-gamma-[35S] and GTP-gamma-[32P] or GTP-alpha-[32P] bound to plasma membranes of rat liver was immunoprecipitated using anti p21V-H-ras. Binding was enhanced approximately 2-fold by incubation with an exogenous electron acceptor, potassium ferricyanide (but not with potassium ferrocyanide), or oxidized ubiquinone10 and was inhibited or unaffected by incubation with reduced pyridine nucleotides (NADH or NADPH) or reduced ubiquinone10. The results suggest a mechanism of guanine nucleotide exchange that is responsive to oxidation-reduction. PMID- 8422248 TI - Protein kinase C phosphorylates Ser152, Ser156 and Ser163 but not Ser160 of MARCKS in rat brain. AB - MARCKS is one of the major physiological substrates of PKC and was reported to be phosphorylated by PKC at 4 serine residues that are within the CaM-binding region (Graff et al., J. Biol. Chem. 264, 11912, 1989). Using MARCKS from rat brain and a synthetic peptide of 25 amino acids containing all 4 of the serine residues, we investigate the differences in phosphorylation by PKC isozymes I, II and III. Tryptic peptide analysis of PKC phosphorylated MARCKS or peptide, we found 32P was in peptides of (K)S152FK, (R)FS156FK and LS160GFS163FK. Further digestion of LSGFSFK with alpha-chymotrypsin revealed that 32P incorporation occurred only at Ser163 but not at Ser160. The initial rates and stoichiomatry of phosphorylation of Ser152 and Ser156 were twice as those of Ser163 using either one of the three PKC isozymes. These results indicate that in vitro, PKC phosphorylates MARCKS only at three sites, but not at Ser160 as that reported previously, and there was no preferential phosphorylation of MARCKS by either PKC isozyme I, II or III. PMID- 8422249 TI - Rational design and expression of a heparin-targeted human superoxide dismutase. AB - In order to improve the therapeutic effectiveness of human Cu,Zn superoxide dismutase (HSOD) by targeting it to cell surfaces and increasing its circulatory half-life, we have designed and expressed a heparin-binding derivative of HSOD. This design was based on the idea that structurally independent protein units, HSOD and the heparin-binding A+ helix from protein C inhibitor, could be combined with a carefully chosen linker, GlyProGly, to form a stable, bifunctional protein. The chimeric HSOD-GlyProGly-A+ protein was expressed and secreted to the periplasm of E. coli and had normal SOD activity. HSOD-GlyProGly-A+ had a significantly increased retention time relative to wild-type HSOD on a heparin affinity column, indicating that it was successfully targeted to heparin, and this binding was maintained at physiological ionic strength. When administered to mice, HSOD-GlyProGly-A+ had a half-life of approximately 15 minutes, twice that of wild-type HSOD. Our rational design approach should be generally applicable to the creation of bifunctional chimeric molecules. PMID- 8422250 TI - A new and rapid method for visualising DNA replication in spread DNA by immunofluorescence detection of incorporated 5-iododeoxyuridine. AB - We have developed a new and rapid immunofluorescent method for visualisation of replicated regions on fixed DNA fibers. Using this method we have found in 5 fluorodeoxyuridine (FrdU)-blocked human cells distinct replication units covering about 60 kb of DNA and corresponding in size to single replicons or chromatin loops. Our results also suggest that nonadjacent replicons within a replicon cluster may be activated after FrdU arrest and that the method may be adapted for localization on fibers of specific DNA sequences. PMID- 8422251 TI - Protein kinase C inhibits the Ca(2+)-activated K+ channel of cultured porcine coronary artery smooth muscle cells. AB - The effect of protein kinase C (C-kinase) on the Ca(2+)-activated K+ channel (KCa channel) was studied in cultured smooth muscle cells from porcine coronary artery by the patch-clamp technique. In cell-attached patches, bath application of phorbol 12-myristate 13-acetate (PMA, 1 microM), a C-kinase activator, significantly decreased the open probability of the activated KCa-channel in the presence of the calcium ionophore A23187 (20 microM), which increases intracellular Ca2+. This decrease in the open probability was reversed by subsequent application of staurosporine (1 nM), a C-kinase inhibitor. Application of 1-oleoyl-2-acetylglycerol (OAG, 30 microM) or 1,2-dioctanoylglycerol (DG8; 30 microM), activators of C-kinase, also inhibited KCa-channel activation by A23187, and these inhibitions were also reversed by staurosporine. PMA (1 microM) also inhibited KCa-channel activation by dibutylyl cyclic AMP (db-cAMP, 2 mM) or caffeine (30 mM). In inside-out patches, bath application of the C-kinase fraction from rat brain in the presence of ATP (1 mM) and PMA (1 microM) markedly inhibited the KCa-channel. These results indicate that activation of C-kinase inhibits the KCa-channel and may cause membrane depolarization and vascular contraction. PMID- 8422252 TI - Angiotensin II receptors in rabbit vascular grafts. AB - New Zealand White rabbits were used as a model of autologous venous bypass grafting. A section of jugular vein was sutured into the divided common carotid artery. The animals were euthanized one month after surgery. At that time histopathology showed myointimal thickening of the graft segments. Angiotensin II receptor assays showed that the AT1 receptor was present in the grafts, concentrated in the intima. These localized receptors were shown to be similar to the prototypic vascular AT1 receptor through the use of the specific antagonist DuP 753. PMID- 8422253 TI - Reductive cleavage of the disulfide bonds of the collagen IV noncollagenous domain in aqueous sodium dodecyl sulfate: absence of intermolecular nondisulfide cross-links. AB - The subunits of the collagen IV hexameric, noncollagenous NC1 domain obtained from bovine aorta, glomerular basement membrane, alveolar basement membrane and placental basement membrane are predominantly dimers. A large fraction of the dimers had been thought to be linked by nondisulfide bonds because they were resistant to cleavage by 5% (v/v) 2-mercaptoethanol, 2% (w/v) SDS, at 100 degrees C. However, if an unusually high concentration of 2-mercaptoethanol, e.g., 40% (v/v), is used, complete conversion of dimers into monomers is achieved, indicating the lack of intersubunit nondisulfide cross-links. Electrophoresis patterns indicate that some of the intermolecular disulfide bonds of the dimers are more resistant to reduction in aqueous SDS than are some of the intramolecular disulfide bonds. PMID- 8422254 TI - Preparation of anti-phosphoserine and anti-phosphothreonine antibodies and their application in the study of insulin- and EGF-induced phosphorylation. AB - We prepared antibodies against phosphoserine (P-Ser) and phosphothreonine (P-Thr) by immunizing rabbits with P-Ser or P-Thr conjugated to bovine serum albumin. The antibodies (anti-P-Ser and anti-P-Thr) were purified using P-Ser or P-Thr affinity columns. Anti-P-Thr was highly specific for P-Thr, while anti-P-Ser showed weak cross-reactivity with P-Thr. We showed that these antibodies can immunodetect serine/threonine phosphorylated insulin and epidermal growth factor (EGF) receptors and several proteins which are phosphorylated on serine/threonine residues in response to insulin or EGF stimulation. The antibodies will certainly provide a good tool for discovering novel kinases and substrates involved in signal transduction. PMID- 8422255 TI - Molecular cloning of the smaller subunit(P52) of rat liver mitochondrial processing protease. AB - A cDNA encoding the smaller subunit (P52) of mitochondrial processing protease was isolated from a rat liver cDNA library, using cDNA fragment for yeast MAS1 as the probe. The deduced amino acid sequence is highly homologous to those of PEP from Neurospora crassa and MAS1 from Saccharomyces cerevisiae. After in vitro transcription and translation, the precursor peptide was imported into isolated rat liver mitochondria and processed to its mature form. PMID- 8422256 TI - Occurrence of 2-O-methyl-N-(3-deoxy-L-glycero-tetronyl)-D-perosamine (4-amino-4,6 dideoxy-D-manno-pyranose) in lipopolysaccharide from Ogawa but not from Inaba O forms of O1 Vibrio cholerae. AB - A structural study by GC-MS, methylation analysis, and 1H and 13C NMR was carried out on alpha (1-->2)-linked linear N-(3-deoxy-L-glycero-tetronyl)-D-perosamine homopolymer constituting the O-polysaccharide chain of lipopolysaccharide from O1 Vibrio cholerae Ogawa and Inaba O forms. Occurrence of 2-O-methyl-N-(3-deoxy-L glycero-tetronyl)-D-perosamine was demonstrated at the non-reducing terminus of the perosamine-homopolymer of lipopolysaccharide from the Ogawa O form in contrast to the presence of N-(3-deoxy-L-glycero-tetronyl)-D-perosamine at the nonreducing terminus for the Inaba O form. PMID- 8422257 TI - Sequence-specific binding of HMG-I (Y) to the proximal promoter of the gp91-phox gene. PMID- 8422258 TI - Calcium-independent activation of protein kinase C by the dianionic form of phosphatidic acid. AB - Phosphatidic acid in the form of small unilamellar vesicles has a dissociation constant of about 8.3 as determined by 31P nuclear magnetic resonance (NMR) spectroscopy. The activation of protein kinase C (PKC) by monovalent phosphatidic acid or phosphatidylserine occurs only in the presence of Ca2+. However, PKC activity on membranes of divalent anionic phosphatidic acid is independent of Ca2+ concentration. PMID- 8422259 TI - The calmodulin antagonist W-7 depletes intracellular calcium stores in FRTL-5 thyroid cells. AB - Incubating Fura 2 loaded thyroid FRTL-5 cells with the calmodulin inhibitor W-7 decreased the ATP-evoked increase in intracellular free calcium. In addition, pretreatment of the cells with W-7 decreased both the thapsigargin-evoked release of sequestered calcium and the entry of extracellular calcium. Studies with 45Ca2+ showed that W-7 decreased the amount of sequestered calcium in the cells. Furthermore, after stimulating with ATP, the ability to sequester calcium was decreased in cells treated with W-7 compared with control cells. The results suggest that calmodulin is necessary for the signal-transduction system in FRTL-5 cells, and may be especially important in maintaining functional intracellular calcium stores in the cells. PMID- 8422260 TI - An explanation for the disparate effects of synthetic peptides corresponding to human follicle-stimulating hormone beta-subunit receptor binding regions (33-53) and (81-95) and their serine analogs on steroidogenesis in cultured rat Sertoli cells. AB - We have recently reported that synthetic peptide amides corresponding to regions of human FSH beta-subunit, hFSH-beta-(33-53) and hFSH-beta-(81-95), bind to receptor and stimulate estradiol biosynthesis by cultured rat Sertoli cells. Because of experimental difficulties caused by the presence of free sulfhydryl groups in these peptides, synthetic analogs were prepared in which all Cys residues were replaced with Ser. These analogs, [Ser-51]-hFSH-beta-(33-53) and [Ser-82, 84, 87, 94]-hFSH-beta- (81-95), also bound to receptor but did not stimulate estradiol biosynthesis by cultured rat Sertoli cells. In order to explain this observation, we compared the effects of hFSH-beta-(33-53) and hFSH beta-(81-95) and their Ser analogs on another recently recognized effect of FSH in Sertoli cells, namely its ability to promote influx of extracellular calcium. We and others have shown that estradiol biosynthesis by these cells is markedly decreased in the presence of high intracellular calcium. Cys-containing hFSH-beta (33-53) and hFSH-beta-(81-95) did not increase influx of extracellular calcium over basal levels, whereas [Ser-51]-hFSH-beta-(33-53) and [Ser-82, 84, 87, 94] hFSH-beta-(81-95) induced 2.8- and 1.8-fold increases, respectively. Cellular cAMP and estradiol biosynthesis in response to each Ser-substituted peptide were not significantly different from basal levels. Thus, the explanation for the observed disparate effects of Cys and Ser analog peptides on estradiol biosynthesis may be related to the ability of the Ser peptides to stimulate calcium entry but not cAMP accumulation in cultured rat Sertoli cells. PMID- 8422261 TI - D-erythro-sphingosine lowers 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in Chinese hamster ovary cells. AB - D-Erythro-sphingosine lowered the levels of HMG-CoA reductase activity in CHO-K1 cells. Significant suppression of reductase activity was observed at 5 microM, 10 microM, and 15 microM concentrations of the sphingolipid base and approximately 50% lowering was found at 10 microM. In contrast, L-threo-sphingosine had no effect on the levels of reductase activity under the conditions studied. Direct addition of D-erythro-sphingosine, at concentrations up to 100 microM, to rat liver microsomes had no effect on the levels of HMG-CoA reductase activity. The concentrations of D-erythro-sphingosine required to lower reductase activity in CHO-K1 cells correspond to reported levels of sphingosine in mammalian cells. PMID- 8422262 TI - Evolution of the ets gene family. AB - Over the past few years a variety of genes have been described whose protein products share similarity with that of the c-ets-1 proto-oncogene, the cellular counterpart of the v-ets oncogene of the avian E26 retrovirus. This so-called "ets family" of transcription factors includes at least a dozen members present in several organisms. We have questioned the common evolutionary origin of these various gene products. By constructing phylogenetical trees with different methods, we show that the ets family is very ancient since the duplication of the various groups of ets related proteins occurred before the Arthropods/Vertebrates split (ca. 500 million years). PMID- 8422263 TI - Effect of H1 histone isoforms on the methylation of single- or double-stranded DNA. AB - The loosely and tightly bound H1 histone isoforms were shown to exert, on the in vitro methylation of linker DNA in H1-depleted oligonucleosomes, inhibitory or activating effects respectively similar to those previously shown in the methylation of Micrococcus luteus dsDNA. When assayed on the enzymic methylation of Micrococcus luteus ssDNA, addition of the tightly bound one resulted in a stimulation similar to that exerted on double-stranded bacterial DNA or on linker DNA from mammalian chromatin, while the loosely bound isoform had no effect whatsoever. The transformation of the "typical" loosely bound H1 isoform into its tightly bound counterpart can be visualized as being an essential event in the modulation of DNA methylation process in eukaryotic chromatin. PMID- 8422264 TI - Genetic exclusion of apo-B gene in recessive abetalipoproteinemia. AB - Abetalipoproteinemia is a recessive genetic disorder of unknown origin, which is characterized by absence of circulating apo-B-containing lipoproteins, malabsorption of intestinal fat, and degenerative neurological and retinal lesions. In this study, four families were analysed for genetic linkage between the abetalipoproteinemia phenotype and the apo-B genotype determined from polymorphisms of XbaI, MsPI, EcoRI and PvuII restriction sites and that of the 3' minisatellite of the apo-B gene. The results definitively exclude mutation of the apo-B gene as a causal factor of abetalipoproteinemia in three families. Consanguinity of the parents in the fourth family made genotyping less conclusive. PMID- 8422265 TI - A survey of retirement planning by Texas psychiatrists. AB - Forty-six percent (274 of 600) of the members of the Texas Society of Psychiatric Physicians responded to a mailed survey that addressed retirement issues. Forty seven percent of respondents were 60 to 69 years of age. Only 12% were female. Ninety-one percent were white, with 86% of respondents married. Only 24% of the sample (66 of 274) were retired at the time of the survey. Although engaged in more than one area of professional activity, 71% were involved in private practice, with 51% practicing in a private office. The major reasons for retiring were "a full life and ready to change pace" (52%) and "retired to pursue other interests" (25%). Twenty percent retired because of physical illness. A mixed financial package formed their retirement package, which included social security (80%), personal savings (73%), Keogh (64%), and state/US government monies (39%). Retired psychiatrists were interested in receiving additional information on successful patterns of retirement (21%), maintaining emotional and physical health (17%), and material on the problems and trials of retirement (15%). Leisure-time activities were varied, and most psychiatrists reported two to four activities. According to 39% of the sample, retirement should be discussed during medical school, 46% indicated that it should be discussed during residency training, and 80% selected Continuing Medical Education courses. PMID- 8422266 TI - Combined use of labetalol and nifedipine in controlling the cardiovascular response from ECT. AB - Serious cardiac complications have been reported to occur in elderly depressed patients during a course of electroconvulsive therapy (ECT). As a result, cardiac medications are being used more often to dampen the cardiovascular response that occurs during an ECT treatment. Specifically, labetalol (a mixed alpha- and beta blocker) has been shown to effectively control the heart rate during ECT. However, on occasion, patients may still exhibit sustained elevations of blood pressure during ECT when receiving labetalol. The optimum clinical management of these patients is unclear. The authors report on the safety and efficacy of combining nifedipine with labetalol to control blood pressure during ECT in ten elderly patients whose blood pressures were not adequately controlled by labetalol alone. No adverse effects were noted, nor did nifedipine appear to shorten seizure duration. PMID- 8422267 TI - Presbyophrenia: a possible subtype of dementia. AB - Presbyophrenia is characterized by memory impairment, disorientation, confabulation, hypomanic features, and a preserved social facade. These occur in the absence of prior history of alcoholism or affective illness. We present three cases with presbyophrenia and suggest that the syndrome is a recognizable subtype of dementia, possibly related to disruption of aminergic pathways in frontal and subcortical structures. PMID- 8422268 TI - Risk factor and behavioral differences between vascular and Alzheimer's dementias: the pathway to end-stage disease. AB - Differences between vascular dementia and Alzheimer's dementia may be present in both risk factors and in behavioral manifestations. Behavioral distinctions may be apparent only at particular stages of the disease process, but comparisons at different stages require a large number of subjects. The California Alzheimer Disease Diagnostic and Treatment Center Program has collected data on a large number of subjects with dementia. We examined differences in risk factors and in behaviors in 502 subjects with vascular dementia and 810 subjects with probable Alzheimer's dementia. With respect to risk factors, vascular dementia subjects were more likely to have a history of general anesthesia. We found no difference in history of closed head trauma, family history of dementia, or cigarette smoking, suggesting that these factors are less likely to have a disease-specific etiologic role. With respect to behaviors, subjects with vascular dementia were more likely to have depression in all stages of dementia. Subjects with Alzheimer's dementia were more likely to exhibit wandering in the middle or late stages. Subjects were similar in five other behavioral factors in the three stages studied. We conclude that general anesthesia is a risk factor for vascular dementia and that depression is more prevalent in vascular dementia, while wandering is more characteristic of Alzheimer's dementia. PMID- 8422269 TI - Brain oxidative energy and related metabolism, neuronal stress, and Alzheimer's disease: a speculative synthesis. AB - A reduction in the cerebral metabolic rate of glucose is one of the most predominant abnormalities generally found in the Alzheimer brain, whereas the cerebral metabolic rate of oxygen is diminished only slightly or not at all at the beginning of this dementive disorder. From the cerebral metabolic rates of oxidized glucose and oxygen, the cerebral adenosine triphosphate (ATP) formation rate was calculated in incipient early-onset, incipient late-onset, and stable advanced dementia of the Alzheimer type (DAT). A reduction in ATP formation by various amounts was found, ranging from at least 7% in incipient early-onset DAT, from around 20% in incipient late-onset DAT, and from 35% up to more than 50% in stable advanced dementia. The cerebral diminution in energy availability, along with a loss of functionally important amino acids, ammonia toxicity, supposed membrane damage, dysregulation of Ca2+ homeostasis, and glycogen accumulation in the incipient stages of DAT are assumed to be stress-related abnormalities capable of inducing the formation of heat shock proteins. These events may lead to an enhanced generation of amyloid precursor protein in earlier states of DAT. If abnormally cleaved, amyloid A4 protein may be produced in increased amounts. From the results discussed in this article it is deduced as a speculative synthesis that perturbations in brain oxidative energy and related metabolism may precede the generation of amyloid precursor protein and the formation of plaques in the brain affected by incipient DAT. PMID- 8422270 TI - Weight loss in Alzheimer's disease. AB - In order to determine whether weight loss is a core symptom or secondary manifestation of Alzheimer's disease, we analyzed weight change in 81 outpatients with Alzheimer's disease. During a mean 2.9 years of observation, the mean absolute weight loss was 1.9 +/- 0.8 kg, and the mean weight loss per year was 0.7 +/- 0.3 kg. Although statistically significant (P < .05), the mean absolute weight loss was only 3% of initial body weight. Data analyses revealed that weight change was significantly (P < .0001) correlated with decreased independence in self-feeding. Weight change was not related to duration of dementia, clinical measures of dementia severity, psychiatric symptoms, or medications. These observations support the view that weight loss is a consequence of Alzheimer's disease and does not reflect specific brain lesions. PMID- 8422271 TI - [Progress report for the period 1 January 1990--31 December 1991]. PMID- 8422272 TI - Molecular cloning of myomodulin cDNA, a neuropeptide precursor gene expressed in neuron L10 of Aplysia californica. AB - Screening of an abdominal ganglion cDNA library of Aplysia californica with a synthetic oligonucleotide based on the myomodulin peptide sequence has been used to identify and characterize a cDNA clone expressed in interneuron L10. The complete sequence of 1,534 nucleotides in length shows an open reading frame of 370 amino acids that encodes a 42-kD pre-propeptide. Proteolytic cleavage of the precursor potentially gives rise to 10 copies of myomodulin A, single copies of 6 myomodulin-related peptides, and other unrelated sequences. Southern blot analysis of sperm DNA shows that the haploid Aplysia genome contains only one copy of the gene. RNA blot analyses of central nervous system (CNS) mRNA show that the myomodulin gene is expressed in all major ganglia and that a single transcript of around 1,600 nucleotides can be detected in pooled CNS mRNA, suggesting that the same sequence is transcribed in each ganglia. Nucleotide sequences of partial cDNA clones isolated from a cerebral ganglion and a CNS cDNA library are identical to the abdominal ganglion sequence, further suggesting that the same myomodulin gene is expressed in other ganglia. PMID- 8422273 TI - Promoter elements and transcriptional regulation of the acetylcholinesterase gene. AB - The 5' region of the acetylcholinesterase gene from the electric ray Torpedo californica has been cloned and its cap site identified. The 5' untranslated region is divided into two exons where a small exon extending between bp -22 to 60 is alternatively spliced. Cap sites are defined at two positions, bp -138 and 143. Twenty-one base pairs 5' of the -143 cap site a repeating TATA sequence is found. Further upstream in the gene consensus sequences for Sp1, AP1, and AP2 factors are evident. The promoter region of the acetylcholinesterase gene enhances transcription of a luciferase reporter gene transfected into C2 myoblasts. However, increased transcription was not evident after C2 myoblasts were induced to form myotubes. Cotransfection of this construct with c-Jun (AP1) and AP2 expression vectors shows marked increases of transcription rates in HepG2 and C2 cells. Protein kinase A elicited regulation of expression is also evident in quail fibroblasts. In gel retardation experiments both recombinant c-Jun (AP1) and AP2 proteins bind to the appropriate Torpedo sequences. Cellular extracts from the Torpedo electric organ exhibit AP2 binding activity. Thus, although all facets of specific regulation expected upon differentiation of mammalian muscle cells were not evident, the 5'-flanking region from the Torpedo AChE gene contains consensus sequences and functional promoter elements typical of mammalian nerve and muscle systems. PMID- 8422274 TI - A human putative lymphocyte G0/G1 switch gene homologous to a rodent gene encoding a zinc-binding potential transcription factor. AB - G0S24 is a member of a set of genes (putative G0/G1 switch regulatory genes) that are expressed transiently within 1-2 hr of the addition of lectin or cycloheximide to human blood mononuclear cells. Comparison of a full-length cDNA sequence with the corresponding genomic sequence reveals an open reading frame of 326 amino acids, distributed across two exons. Potential phosphorylation sites include the sequence PSPTSPT, which resembles an RNA polymerase II repeat reported to be a target of the cell cycle control kinase cdc2. Comparison of the derived protein sequence with those of rodent homologs allows classification into three groups. Group 1 contains G0S24 and the rat and mouse TIS11 genes (also known as TTP, Nup475, and Zfp36). Members of this group have three tetraproline repeats. Groups 1 and 2 have a serine-rich region and an "arginine element" (RRLPIF) at the carboxyl terminus. All groups contain cysteine- and histidine rich putative zinc finger domains and a serine-phenylalanine "SFS" domain similar to part of the large subunit of eukaryotic RNA polymerase II. Comparison of group 1 human and mouse genomic sequences shows high conservation in the 5' flank and exons. A CpG island suggests expression in the germ line. G0S24 has potential sites for transcription factors in the 5' flank and intron; these include a serum response element. Protein and genomic sequences show similarities with those of a variety of proteins involved in transcription, suggesting that the G0S24 product has a similar role. PMID- 8422275 TI - Molecular characterization of the cDNA coding for translation elongation factor-2 of pathogenic Entamoeba histolytica. AB - To investigate the humoral immune response of patients with amoebic dysentry against Entamoeba histolytica, immunoglobulin G (IgG)-immunopositive cDNA clones from the pathogenic strain SFL-3 were examined. A large part of the IgG-positive cDNA clones obtained with one serum encoded highly conserved intracellular proteins. A clone was found that was homologous to translation elongation factor 2 (EF-2). Sequence analysis of the EF-2 cDNA showed 63.6% amino acid sequence identity with the human homologue. The deduced protein sequence has a length of 840 amino acid residues with a molecular mass of 93.3 kD. The 3' and 5' untranslated regions of the mRNA are relatively short as shown for other genes of E. histolytica. A genomic clone was used to analyze the region upstream of the translation initiation codon. The codon distribution of EF-2 and other published E. histolytica sequences reflects the high A/T content. The codons for different amino acids are biased to a widely differing extent. PMID- 8422276 TI - Post-translational processing of barley beta-glucan endohydrolases in the baculovirus-insect cell expression system. AB - Two cDNAs encoding barley (1-->3,1-->4)-beta-glucanase (EC 3.2.1.73) isoenzymes EI and EII have been expressed in Spodoptera frugiperda (Sf9) cell cultures using the baculovirus AcNPV vector. Modifications to both the 5' and 3' ends of the cDNAs were required before satisfactory levels of expression were obtained. The modified cDNAs directed high levels of (1-->3,1-->4)-beta-glucanase expression in the Sf9 insect cell cultures, with yields of approximately 10 mg/liter of isoenzyme EI (expEI) and 15 mg/liter of isoenzyme EII (expEII). Amino acid sequence analyses showed that the expressed enzymes were processed correctly at their amino termini. However, affinity chromatography of the isoenzyme expEII on concanavalin-A (conA)-Sepharose indicated that, although the enzyme is glycosylated, the structures of the carbohydrate chains differ from those of the native enzyme. When a cDNA encoding the homologous barley (1-->3)-beta-glucanase (EC 3.2.1.39) isoenzyme GII was expressed in insect cells, aberrant amino terminal processing of the nascent polypeptide was sometimes observed. The forms with incompletely removed signal peptides retained their substrate specificity, but exhibited slightly reduced catalytic efficiency, altered chromatographic behavior, and reduced stability at elevated temperatures. The results show that high levels of expression of recombinant plant proteins can be obtained in insect cells, but they emphasize the need to characterize thoroughly the products that are expressed in the heterologous insect cell system before comparisons are made with the native enzyme or with engineered enzyme mutants. PMID- 8422277 TI - Pancreatic cancer: how can we progress? PMID- 8422278 TI - Quality assurance in cancer treatment. Report of a Working Party from the European School of Oncology. PMID- 8422279 TI - The impact of received dose intensity on the outcome of advanced ovarian cancer. AB - It has been demonstrated that the prognosis of ovarian cancer is influenced by the dose intensity of cytotoxic treatment. The impact of received dose intensity of platinum-based combination chemotherapy on disease outcome was analysed in 226 stage III-IV ovarian cancer patients entered into two prospective randomised trials. All patients received either cisplatin or carboplatin and cyclophosphamide with or without doxorubicin for six courses after primary surgery. The impact of the received dose intensity of each drug (RDI), the average received dose intensity of the treatment regimen (ARDI) and the relative total drug dose (RTD) on progression-free survival (PFS) and survival were analysed. In the 198 patients receiving the full six courses of treatment, RDI of cisplatin or carboplatin, ARDI and RTD were > 0.76 in 74.2, 61.1 and 65.1% of cases, respectively. Although the differences were not significant, pathological complete response was more frequently observed in the group of patients with ARDI < 0.75, whereas the partial response rate was higher in the ARDI > or = 0.76 group. Median survival and PFS were 19 and 13 months; 22 and 10 months; 23 and 13 months for the groups of patients receiving chemotherapy at a ARDI of < 0.75, > or = 0.76-0.99 and > 1.00, respectively (P = not significant). It appears that modest dose modifications and brief treatment delays during first-line platinum based chemotherapy do not affect response rate, survival and PFS in advanced ovarian cancer patients. PMID- 8422280 TI - Neuroimmunotherapy of advanced solid neoplasms with single evening subcutaneous injection of low-dose interleukin-2 and melatonin: preliminary results. AB - On the basis of the demonstrated existence of immunoneuroendocrine interactions and on the previously observed synergistic action between the pineal hormone melatonin (MLT) and interleukin-2 (IL-2), we have designed a neuroimmunotherapeutic combination consisting of low-dose IL-2 and MLT in the treatment of advanced solid neoplasms. The study included 24 patients with advanced solid tumours (non-small cell lung cancer 9; colorectal cancer 7; gastric cancer 3; breast cancer 2; cancer of pancreas 1; hepatocarcinoma 1; unknown primary tumour 1), 21 of whom showed distant organ metastases. Not all patients responded to previous chemotherapies, or had tumours for which no standard therapy was available. Moreover, not all patients were able to tolerate IL-2 immunotherapy at the conventional doses. IL-2 was given subcutaneously at a dose of 3 x 10(6) U/day at 8:00 p.m. for 6 days/week for 4 weeks. MLT was given orally at a dose of 50 mg at 8:00 p.m. every day, starting 7 days before IL-2 injection. In non-progressed patients, a second cycle was given after a 21-day rest period. A partial response was seen in 3/24 patients (lung 2; stomach 1; duration: 11, 4, 4 months, respectively). Moreover, a minimal response (duration: 8+ months) was seen in 1 lung cancer patient. Stable disease was obtained in 14/24 patients (median duration: 6+ months), while the remaining 6 patients progressed. An improvement in performance status was seen in 7/24 patients. No important toxicity was observed. Mean eosinophil and lymphocyte levels significantly increased during the immunotherapy, and their rise was significantly higher in patients with response or stable disease than in those with progressive disease. These preliminary results show that neuroimmunotherapy with low-dose IL-2 and the pineal hormone MLT is a biologically active and well tolerated strategy, capable of determining an apparent control of tumour growth in patients with advanced solid neoplasms, for whom no standard effective therapy is available. PMID- 8422281 TI - Prednisolone, cytosine arabinoside, lomustine (CCNU), etoposide and thioguanine (PACET) combination chemotherapy for relapsed or refractory non-Hodgkin lymphoma. AB - 27 patients with relapsed/refractory non-Hodgkin lymphoma (NHL) received combination chemotherapy with prednisolone, cytosine arabinoside, lomustine (CCNU), etoposide and thioguanine (PACET). 25 patients are evaluable for response. 7 (26%) obtained a complete response and one (4%) a partial response. The median survival for the entire group was 6 months. 2 patients are currently alive without disease, 1 of whom has received further therapy. The regimen was intensely myelosuppressive, but was well tolerated. The complete response rate and median survival figures are comparable to previous studies of salvage therapy confirming the poor prognosis for relapsed NHL and emphasising the need for prospective randomised studies. PMID- 8422282 TI - Neuron-specific enolase in cerebrospinal fluid of patients with metastatic and non-metastatic neurological disease. AB - Neuron-specific enolase (NSE) activities were measured in cerebrospinal fluid (CSF) in 361 patients with various neurological diseases. CSF was collected as part of the diagnostic procedure both in the control group, which consisted of 189 subjects with low back pain, and in the patient group (172 patients). The mean CSF NSE level in 189 control subjects was 7.14 +/- 1.94 micrograms/l. Slight elevations of CSF NSE (> or = 11.0 micrograms/l) were observed in 9 patients with non-malignant diseases and in 2 patients with malignant diseases. The findings of this study indicate that measurement of NSE in CSF cannot be used as an adjunctive diagnostic test for CNS metastases. PMID- 8422283 TI - Activity of cisplatin in adenocarcinoma of the pancreas. AB - The activity of cisplatin in metastatic adenocarcinoma of the pancreas was assessed in 33 patients. Cisplatin was administered in a dose of 100 mg/m2, every 4 weeks. There were 2 complete responses and 5 partial responses (a response rate of 21%). The median duration of response was 5 months (range, 2+ to 15 months). Cisplatin is a modestly active drug in advanced pancreatic cancer. PMID- 8422284 TI - Phase II study of miltefosine (hexadecylphosphocholine) in advanced soft tissue sarcomas of the adult--an EORTC Soft Tissue and Bone Sarcoma Group Study. AB - The EORTC Soft Tissue and Bone Sarcoma Group conducted a phase II study with oral miltefosine at a dose of 50 mg thrice daily in patients with metastatic soft tissue sarcomas. No responses were seen in 18 evaluable patients. Toxicity consisted mainly of nausea/vomiting. It is concluded that oral miltefosine has no activity in soft tissue sarcoma. PMID- 8422285 TI - Serum immunoreactive and bioactive lactogenic hormones in advanced breast cancer patients treated with bromocriptine and octreotide. AB - 6 patients with advanced breast cancer who had failed first and second line endocrine therapies received bromocriptine (1.25-2.5 mg twice daily per os) and octreotide (Sandostatin) via a continuous subcutaneous infusion (200-400 micrograms/24 h) until disease progression. Pre-treatment 24-h profiles of serum lactogenic hormones and their response to standard provocative tests were established and repeated at 2 weeks, and 3 and 6 months (or at tumour progression). Immunoreactive prolactin (ir-PRL), growth hormone (ir-GH) and insulin-like growth factor I (IGF-I) were measured by radioimmunoassay and bioactive lactogenic hormone levels (BLH) were estimated using the Nb2 rat lymphoma cell bioassay. Before treatment all patients showed episodic secretion of ir-PRL, ir-GH and BLH and provocative stimuli resulted in a peak of ir-GH and BLH maximal between 60 and 90 min after injection but no change in ir-PRL. After 2 weeks of treatment, ir-PRL levels were reduced to below the limit of detection in all 6 patients. Peaks of ir-GH and BLH were still apparent, although much reduced. Immunoreactive PRL continued to be profoundly suppressed in 3 of the 4 patients who remained on treatment for 3 to 6 months. Small pulses of ir-GH were still detectable in these patients with which BLH was, again, well correlated. After 2 weeks of treatment, serum IGF-I levels were reduced by 9-54% of the pretreatment values and generally remained suppressed throughout treatment. Clinically, 4 patients did not show disease progression for periods of up to 6 months and side-effects were minimal. PMID- 8422286 TI - Preferential antibody targeting to small lymphoma metastases in the absence of the primary tumour. AB - Targeting of spontaneous liver metastases of the ESb.MP murine lymphoma was achieved with anti-CD2 monoclonal antibody (MAb) 12-15A, which does not react with normal liver tissue. Using quantitative autoradiography on whole body sections of animals that had received a standard dose of 1.1 MBq of 125I-labelled monoclonal antibody, metastases accumulated up to > 90% of the injected dose per gram (id/g). The average uptake of primary tumour lesions was at a low level of 24 Bq/mg (corresponding to 2.2% id/g) because of highly non-uniform accumulation, while metastatic lesions were all above 50 Bq/mg. Uptake was particularly pronounced in animals tested after resection of the primary tumour: 85% of metastases showed levels above 300 Bq/mg, which was the upper limit of uptake in metastases of non-resected animals. These findings demonstrate the potential of the antibody approach with regard to attacking residual metastatic lesions after debulking. PMID- 8422287 TI - Suramin-induced growth inhibition and insulin-like growth factor-I binding blockade in human breast carcinoma cell lines: potentially related events. AB - Suramin, a polyanionic drug used in the treatment of trypanosomiasis and onchocerciasis, inhibits growth factor-induced mitogenesis in several human tumours. We have investigated the effect of suramin on human breast cancer cell lines (HBCCL). By cell counts and thymidine incorporation we found that 50 to 400 micrograms/ml suramin inhibits the proliferation of HBCCL in a dose-dependent and reversible fashion (ID50 approximately 200 micrograms/ml for MCF-7 and MDA-MB 231). Radioreceptor and affinity cross-linking assays showed that suramin was also able to reduce the binding of insulin-like growth factor I (IGF-I) to its receptor (40-50% inhibition at 100 micrograms/ml). Our results indicate that the drug does not affect the IGF-I receptor (IGF-I-R), but binds directly to the IGF I peptide. In conclusion, the strict correlation observed between suramin inhibition of proliferation and IGF-I binding on HBCCL suggests a possible therapeutic role for this molecule as an antineoplastic drug in human breast tumours. PMID- 8422288 TI - MHC class I expression on prelymphomatous and lymphomatous B-cells is not inhibited by an E mu-myc transgene. AB - Increased expression of oncogenes from the myc family has been associated with down-regulation of major histocompatibility complex (MHC) class I molecules. In certain models this has been suggested to contribute to tumour progression. Transgenic mice bearing the cellular myc oncogene coupled to the lymphoid specific immunoglobulin heavy chain enhancer (E mu) develop clonal B lymphoid malignancies early in life. We have asked if expression of such a constitutively activated E mu-myc transgene in BALB/c mice affects MHC class I expression. H-2Kd and Dd expression on prelymphomatous and lymphomatous B-cells as well as newly established pre-B or B lymphoma cell lines derived from E mu-myc transgenic BALB/c mice were analysed. The results reveal no down-regulated or otherwise altered expression of H-2Kd or Dd on any of the cell populations examined. The results are discussed in relation to the myc associated down-regulation of MHC class I molecules observed in other experimental models. PMID- 8422289 TI - In vitro effect of suramin on lung tumour cells. AB - In the search for new therapeutic concepts in lung cancer chemotherapy, suramin, a potential anticancer drug which evades multidrug resistance, was tested in vitro on 25 lung-derived cell lines, either non-tumorigenic cells, or established cell lines from five different tumour types. Suramin treatment resulted in a time and dose-dependent decrease in [3H]thymidine incorporation, except in one adenocarcinoma cell line where DNA synthesis was highly stimulated. [3H]Leucine incorporation was less affected, indicating that suramin acted cytostatically rather than cytotoxically. Our results show that suramin affected DNA synthesis of the different types of lung derived cells, including non-tumorigenic and tumour cell lines, to a similar extent. PMID- 8422290 TI - Oestrogen receptor message in premalignant and normal cervical cells: a methodological study. AB - Normal and abnormal biopsies of the uterine cervix, to a total of 124 samples, have been analysed for the detection of oestrogen receptor (ER) mRNA. The tough fibrous nature of the cervix proved very resistant to disaggregation in the first instance and subsequently, it was difficult to extract good high molecular weight message. This necessitated a systematic study of methodological technique, including two methods of tissue disaggregation and five techniques of extraction of ER mRNA, which in total involved the use of 124 cervical biopsies. The most successful method involved chopping the tissue, then digesting the cells with proteinase K and extracting the nucleic acids in salt and sodium dodecyl sulphate. Using the perfected technique, 16 cervical biopsies obtained at serial intervals from four women did not show any differences in ER mRNA in cervical biopsies either in the presence of oral contraception or histological abnormality. The successful method described will prove valuable for the detection of ER message in human tumours and other tissues of similar nature. PMID- 8422291 TI - Interval cancers and sensitivity in the screening centres of the UK trial of early detection of breast cancer. AB - The incidence rates of interval cancers following a negative breast screen in two screening centres which offered women aged 45-64 annual screening by mammography and/or clinical examination are examined. Sensitivity of screening is estimated by comparing the incidence rate of interval cancers with that expected in the absence of screening, and the results are compared with those from alternative methods of calculating sensitivity. The incidence rate of cancers diagnosed within 12 months of a negative screen by mammography plus clinical examination was reduced by 70% for women aged 45-54, and 84% for women aged 55+. There is no indication from this that sensitivity in the UK trial was substantially lower than in other studies which have achieved larger reductions in mortality. PMID- 8422292 TI - Rheumatoid arthritis as a risk factor for multiple myeloma: a case-control study. AB - This population-based case-control investigation was designed to study the importance of rheumatoid arthritis, other diseases and different types of treatment for the risk of developing multiple myeloma. In total, 275 cases with verified myeloma in northern Sweden were matched to as many control subjects. Information about different diseases, drug use, diagnostic X-ray investigations and radiotherapy was obtained through an extensive questionnaire mailed to all living subjects, i.e. cases and controls, and to the next of kin regarding deceased subjects. The study confirmed a suspected association with both rheumatic diseases in general and rheumatoid arthritis specifically. No other disease gave an increased risk for myeloma, but on the contrary, other diseases were in general more common among the controls. In accordance with this finding, use of medications and diagnostic X-ray investigations were also less common in cases than in controls. The study did, however, give some support to a recent finding that the number of X-ray investigations might be a risk factor for myeloma. Earlier radiotherapy on benign indications was more common in cases, whereas radiotherapy for malignant disease was more common among controls in this study. PMID- 8422293 TI - Prognosis of rectal cancer in France. AB - We studied changes in the prognosis of cancer of the rectum (excluding the rectosigmoid junction) from 1978 to 1986 in the French department of Calvados on the basis of the 616 cases in the cancer registry. Taken as whole, survival has improved slightly with time (P < 0.01), but the improvement is only significant for men (P < 0.02), patients under 70 years (P < 0.01) and patients living in urban areas (P < 0.05). With regard to tumour characteristics, the improvement was significant only for patients with Dukes' stage C tumours at surgery (P < 0.02). To determine the reasons for the improvement in survival, the year of diagnosis and all other prognostic factors were studied in a multivariate model. Diagnostic conditions such as age and tumour stage did not vary from 1978 to 1986; in contrast, the rates of tumour resection and adjuvant radiation therapy increased, possibly explaining at least part of the improvement, particularly for patients with Dukes' stage C tumours. PMID- 8422294 TI - Activities of antioxidant enzymes and lipid peroxidation in endometrial cancer. AB - Antioxidant enzyme activities and lipid peroxidation were analysed in normal endometrium and endometrial cancer tissues from Finnish and Japanese patients. The catalase and glutathione peroxidase activities of normal endometrium were significantly lower in Finns than in Japanese. Lipid peroxidation was slightly higher in endometrial cancer as compared with normal endometrium both in the Finns and in the Japanese. When cancer tissues were compared with normal endometrium both in Finns and Japanese the activity of superoxide dismutase was significantly lower in cancer tissue than in normal endometrium. In Finns glutathione S-transferase activity was also lower in endometrial cancer tissue than in normal endometrium, and a similar tendency was also found in Japanese. This study suggests that endometrial cancer tissue is associated with an impaired enzymic antioxidant defence system. PMID- 8422295 TI - Regional lung cancer death rates unrelated to smoking? The case of The Netherlands. AB - It has been observed in various countries that regional variation in lung cancer mortality can hardly be explained by differences in current smoking. This paper addresses the question of whether mortality variation within. The Netherlands in 1980-1984 is due to differences in past instead of current smoking. A first indication of the role of past smoking is that, within male birth cohorts, regional mortality patterns have been very stable for over 30 years. Reliable data on smoking in 1972 explain 40% of the mortality variation among women, but only 2% of that among men. A crude indicator on smoking in 1930 explained 43% of the mortality variation among men aged 75+ years (correlation = 0.66). The lack of a relationship with smoking in 1972 appears to be due to a radical change in regional smoking differences, which caused smoking in 1972 to be unrelated to smoking in 1930; long time lags, so that these changes were not yet followed by changes in regional mortality differences. It is concluded that the explanation of regional lung cancer death rates sometimes has to go far back in time. Studying determinants of lung cancer by means of regional analyses requires a more detailed control for smoking history than has been usual. PMID- 8422296 TI - Black (air-cured) and blond (flue-cured) tobacco and cancer risk II: Pharynx and larynx cancer. AB - Two case-control studies have examined the relationship between black or blond tobacco smoking and the occurrence of pharynx or larynx cancer. The first study was carried out in several European countries. Tobacco smoking was found to be associated with higher risks for supraglottic and epilarynx cancer localisations than for pharynx, glottic and subglottic localisation. In all localisations, risk was twice as high again in users of black tobacco after adjusting for alcohol and for lifetime average daily dose of tobacco. The other study was carried out in Uruguay. After taking into account age, age at start of smoking, duration of smoking, years since stopping smoking and filter use, risks were found to be higher in black tobacco smokers than in blond tobacco smokers. All known studies which have been performed in countries where blond tobacco is generally smoked showed lower risks even when adjusted for alcohol. Use of black tobacco appears to be associated with higher risks of cancer of the pharynx and larynx than blond tobacco use. PMID- 8422297 TI - The assessment of body image in cancer patients. AB - It is well recognised that cancer treatment can have a negative impact on body image, and this has proved to be an important outcome variable in treatment comparisons, such as surgery for breast cancer. However, there has been a good deal of variation in the way in which dissatisfaction with body image has been assessed, making comparison of results difficult. Some scales or subscales appear promising but lack the rigorous testing required to confirm their accuracy and reliability. This paper reviews the techniques and questionnaires that have been used for the evaluation of body image and highlights their strengths and weaknesses with respect to their use with cancer patients. At the present time, no single scale stands out as the ideal measure, but a pool of items can be generated from recent research, which merit future evaluation. PMID- 8422298 TI - Feasibility trial of a combination of vinorelbine, ifosfamide, fluorouracil and folinic acid (VIF regimen) in advanced urothelial cancer. PMID- 8422299 TI - Long-term complete remissions in patients with disseminated melanoma treated by fotemustine and dacarbazine. PMID- 8422300 TI - Extrapyramidal reaction associated with ondansetron. PMID- 8422302 TI - Phase II trial with ifosfamide in pancreatic cancer. PMID- 8422301 TI - Phase II and pharmacokinetic study of fotemustine in inoperable colorectal cancer. PMID- 8422303 TI - Triple malignant neoplasms in a patient with adult T-cell leukaemia. PMID- 8422304 TI - Treatment of advanced neuroblastoma. PMID- 8422305 TI - Treatment of advanced neuroblastoma. PMID- 8422306 TI - High interest. PMID- 8422307 TI - The future of vocational training. PMID- 8422308 TI - The title 'doctor'. PMID- 8422309 TI - Mike Grace talks to Dave Geddes. AB - David Geddes is a general practitioner in Portland in Dorset. After seven years in the Royal Army Dental Corps he has been in general practice since 1983 and in his present practice since 1986. His practice is one-third private and two-thirds exempt patients. He was one of the originators of the 'Forum for the Future of Dentistry', and played a prominent role in organising a meeting with Baroness Hooper to try and demonstrate the difficulties local practices were having with the funding of the new contract in November 1991. PMID- 8422310 TI - Mercury spillage. PMID- 8422311 TI - Night-grinding. PMID- 8422312 TI - 'Sick dentist scheme'. PMID- 8422313 TI - Re-using disposables. PMID- 8422315 TI - 'GA for tooth extraction in children in London'. PMID- 8422314 TI - Maintaining the airway during the treatment of severe facial injuries. PMID- 8422316 TI - 'A problem with dentures'. PMID- 8422317 TI - The dental status and attitudes of patients at risk from infective endocarditis. AB - Infective endocarditis represents one of the few potentially fatal infections that may be caused in the patient by a dentist. Efforts to reduce the incidence of this disease usually take the form of appropriate antibiotic cover prior to dental treatment, together with the establishment and maintenance of good oral health. This paper is a report of a survey on the dental health and awareness of 81 'at-risk' patients attending a cardiology outpatient clinic. It was found that whilst the patients were apparently well motivated and well informed, a high prevalence of periodontal disease existed in the group. It is concluded that when patients are diagnosed with a cardiac disorder which predisposes to infective endocarditis they should have a dental examination as soon as possible, preferably by a dentist experienced in the treatment of patients with special needs. PMID- 8422318 TI - Effect of mixing speed on mechanical properties of encapsulated glass-ionomer cements. AB - It is well established that the strength of encapsulated dental amalgam is affected by the speed of mixing. An investigation into the effects of mixing speeds on the compressive strength of three encapsulated glass-ionomer cements was carried out. The working and setting time and compressive strength at 24 hours and 7 days were evaluated for the three materials at four mixing speeds. There appeared to be little variation in working and setting times at the different mixing speeds. The early compressive strength of one material, Ketac fil, increased as the speed of mixing increased. The variation in mixing speed did not appear to affect the early compressive strengths of the other two materials. Seven-day compressive strengths of all three materials were not affected by variations in mixing time. PMID- 8422319 TI - Pericoronitis and accidental paracetamol overdose: a cautionary tale. AB - A case of accidental paracetamol overdose in a patient suffering from pericoronitis is described. Self-medication with paracetamol was exacerbated by the prescription of a compound analgesic containing paracetamol by the patient's dental practitioner. The consequent overdose of paracetamol resulted in liver toxicity and acute liver failure. The hazards of accidental paracetamol overdose are discussed and analgesic preparations containing paracetamol described. PMID- 8422320 TI - Dental pay in the 21st century. AB - The current debate on our pay system sparked off by the elegant dissertation by David Watson-James (BDJ, October 24 1992) has been welcomed by those of us in the profession who try to see past the size of our next monthly schedule. Having spent the last eight years as a member of the Representative Board of the BDA and the last four as a member of GDSC, perhaps I may be forgiven for being more than a little cynical when listening to discussions concerning the NHS/GDS and dentists' pay and conditions of (self-)employment. PMID- 8422321 TI - The future of NHS dentistry. AB - The Government has set out clear policy objectives for the development of a health strategy. In the 'Health of the Nation' there is a strong emphasis on preventive care, stringent quality control and a devolution of central responsibility to give greater flexibility and discretion at a local level. PMID- 8422322 TI - Pediatric neurosurgery: guidelines and cost containment. PMID- 8422323 TI - Classification of slit-ventricle syndromes using intracranial pressure monitoring. AB - The term slit-ventricle syndrome appears to have different meanings to different authors. This study proposes a sublcassification of headaches in shunted children based on data obtained from chronic monitoring of intracranial pressure in 7 symptomatic children. Five distinct syndromes are identified: (1) intermittent, extremely low pressure headaches that are analogous to spinal headaches, (2) intermittent proximal obstruction, (3) shunt failure with small ventricles ('normal volume hydrocephalus'), (4) intracranial hypertension with working shunts (hydrocephalic pseudotumor), and (5) headaches unrelated to shunt function. We would prefer to limit the use of the term slit-ventricle syndrome to the triad of intermittent headaches lasting 10-30 min, smaller than normal ventricles on imaging studies, and slow refill of shunt-pumping devices. In other situations, a description relating to the presumed pathogenesis should be used. PMID- 8422324 TI - Neuron-specific enolase and glial fibrillary acidic protein in vitamin-A-induced mouse myeloschisis: an immunohistochemical study. AB - In an effort to establish the appropriate timing of myeloschisis repair, changes in the exposed neural tissue were studied during fetal development. Neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) were examined immunohistochemically in mice with vitamin-A-induced myeloschisis. As in the normally developing lumbosacral spinal cord, NSE was already expressed in the cytoplasm of neurons in the basal plate of the neural plaque in 16-day-old embryos. GFAP became positive at day 17 both in normal embryos and at the outer border of the plaques in dysraphic embryos. Expression of both NSE and GFAP in normal controls was unchanged in intensity and localization during later fetal development. In contrast, the expression of GFAP increased during later development in the neural plaque of dysraphic animals and suggests a progressive gliosis of tissue with the passage of time. The expression of NSE in the plaque did not change during this time. These results suggest that the neural plaque retains intact neurons in the face of progressive gliosis. Moreover, the results suggest that repair of the myeloschisis should start before irreversible changes are established by progressive gliosis. PMID- 8422325 TI - Tethered spinal cord in patients with anorectal and urogenital malformations. AB - We reviewed 26 patients with anorectal or urogenital malformations managed by the neurosurgical service at The Children's Hospital for tethered spinal cord as diagnosed by spinal MRI. There were 13 patients with cloacal exstrophy, 5 with the Vater association, 3 with imperforate anus, and 5 with other complex anomalies. Tethering spinal cord lesions included myelocystocele, lipomyelomeningocele, and filum lipoma. We review the clinical course, management, correlation of MRI and intraoperative findings, and outcome of the patients, as well as the embryology which underlies the association of these malformations. PMID- 8422326 TI - Posterior fossa extradural hematomas in children. AB - Posterior fossa extradural hematomas (PFEHs) are less frequent than supratentorial extradural hematomas. 9 children operated on for PFEH are reported. There were 5 boys and 4 girls aged from 1 to 16 years. The mode of injury was fall in 6 and traffic accident in 3 cases. Diagnosis of PFEH was made by computed tomography (CT) scanning in 8 cases. In 1 patient, who deteriorated suddenly, respiratory arrest developed before surgery, and the patient died in spite of evacuation of the hematoma. CT scanning enables early diagnosis and reveals the associated supratentorial lesions. Prompt surgical evacuation of PFEHs is the treatment of choice. PMID- 8422327 TI - Traumatic optic neuropathy in children: a prospective study. AB - Fifty children with indirect optic nerve injury were prospectively studied over an 8-year period. They constituted 20% of all patients with optic nerve injury managed on a slightly different protocol. Half of these children were injured due to fall from a height, followed by road traffic accidents in 40% of the cases. Forty-three children had unilateral and 7 children had bilateral visual impairment. Five children had definite history of delayed-onset visual involvement, which ranged from 12 to 72 h following injury. Fifty percent of these children had fractured skulls and 14% had optic canal fracture. Visual evoked potentials were performed in 43 patients of which wave formation was normal in 7 and abnormal in 14 patients. The remaining 22 patients had no wave formation. All the patients were managed conservatively and received corticosteroids for a period of 3 weeks. In 7 children an optic nerve decompression was performed between 4 and 6 weeks following injury. Spontaneous visual improvement was observed in 20 (40%) children. In all the patients onset of visual recovery was noticed within 3 weeks. Of the 7 children who had optic nerve decompression, 4 had minimal spontaneous recovery and their vision remained static. All these 4 children benefited by surgery and the vision improved further following the decompression. Three children had no perception of light prior to surgery and none of these patients showed visual improvement following surgery. The present study brings out the clinical profile of children with optic nerve injury and the indications of surgery in these patients. PMID- 8422328 TI - Complications of epilepsy surgery in children and adolescents. AB - The Children's Hospital of Eastern Ontario's Epilepsy Surgery Series of 47 medically intractable children and adolescents was reviewed and analyzed to assess complications of surgical treatment. Complications and rates of incidence are discussed, and compared with the existing literature regarding children, adolescents and adults. No deaths or permanent disabling complications were incurred. Minor morbidity was minimal: 2 (4.2%) sustained permanent but nondisabling complications while an additional 3 (6.4%) incurred transient neurologic deficits which resolved rapidly. Epilepsy surgery in children and adolescents is safe, highly effective, and underutilized. PMID- 8422329 TI - Radical resections of childhood craniopharyngiomas. AB - A series of 27 children with craniopharyngiomas is presented. All the craniopharyngiomas were detected by computed tomography, and radical total resection was attempted. Postoperative evaluations were done by a series of computed tomography and magnetic resonance imaging scans. Twenty patients who had total resection confirmed by surgical and radiographic observations showed no recurrence during the follow-up period of from 1 to 12 years. It is emphasized that total resection using modern diagnostic and surgical methods is the mainstay for childhood craniopharyngiomas. PMID- 8422330 TI - Influence of the apoA-II gene locus on HDL levels and fatty streak development in mice. AB - Previous studies have shown that distal mouse chromosome 1 contains the apolipoprotein AII (apoAII) gene, encoding the second most abundant apolipoprotein in high density lipoproteins (HDLs), as well as a gene termed Ath 1 that controls aortic fatty streak development and HDL cholesterol levels in response to a high-fat, high-cholesterol diet. We report genetic studies confirming that the genes are distinct. Using molecular markers for mouse chromosome 1, we have further mapped the two genes, and our results indicate that they are separated by a minimum of 2 cM. We also report evidence that in mice on a low-fat chow diet, the apoAII gene locus influences HDL cholesterol levels. Thus, statistical analysis of two sets of recombinant inbred strains revealed concordant segregation patterns of HDL cholesterol levels and the apoAII gene locus. The effect of apoAII expression on HDL cholesterol levels was further tested by using a congenic strain that exhibits increased apoAII synthesis in comparison to the background strain. The results support the concept that increased synthesis of apoAII results in increased HDL cholesterol levels. Unexpectedly, increased expression of apoAII appeared to promote rather than retard aortic fatty streak development. PMID- 8422331 TI - Regulation of factor VIIa/tissue factor functional activity in an umbilical vein model. AB - Activation of factor IX in an umbilical vein model was established to result solely from factor VIIa/tissue factor (TF) activity generated within the umbilical vein wall, and the model was then used to study regulation of such extravascular factor VIIa-TF complexes. Vein segments were filled with a reaction mixture containing factor VIIa, Ca2+, a substrate, either [3H]factor IX or [3H]factor X, and a test material. Subsamples were assayed for activation peptide release. Test materials included defibrinated plasma or recombinant protein as a source of TF pathway inhibitor (TFPI), recombinant factor VIIa to 10 times plasma factor VII concentrations, and annexin V. A plasma concentration of TFPI inhibited but did not totally suppress factor VIIa/TF activity. Reducing the TFPI concentration by 50% markedly reduced the inhibition. A 10-fold increase in the factor VIIa concentration in reaction mixtures failed to accelerate factor Xa generation. Annexin V, in contrast to its inhibition of factor VIIa/TF formed with TF reconstituted into mixed phospholipid vesicles, failed to inhibit factor VIIa-TF complexes formed within the vessel wall. We conclude that 1) moderate variation in plasma TFPI concentration or activity may affect TFPI's ability to inhibit factor VIIa/TF activity during hemostasis, 2) a plasma concentration of factor VII suffices to saturate TF sites exposed in a vessel after tissue injury, and 3) the resistance of factor VIIa-TF complexes to inhibition by annexin V suggests that they are formed in the umbilical vein model primarily on cell surfaces. PMID- 8422332 TI - HDLs and alimentary lipemia. Studies in men with previous myocardial infarction at a young age. AB - The plasma concentration, particle size, and chemical composition of high density lipoproteins (HDLs) are associated with the metabolism of triglyceride-rich lipoproteins (TGRLs). During alimentary lipemia there is active exchange of lipids and apolipoproteins between HDL and apolipoprotein B-containing lipoproteins. Whereas HDL has been assigned a protective role against the development of atherosclerosis, alimentary lipemia has been proposed to represent a potentially atherogenic state. We examined plasma HDL concentration, particle size, and composition and their relations to postprandial TGRLs in 32 postinfarction patients and 10 healthy control subjects after intake of a standardized oral fat load of a mixed-meal type. All patients had undergone coronary angiographies in connection with the myocardial infarction and around 5 years thereafter. The plasma HDL cholesterol concentration decreased significantly in response to the oral fat load, particularly in hypertriglyceridemic patients, with a concomitant increase of HDL triglycerides. A limited and reversible yet consistent increase of HDL particle size (1-2%) was seen 6 hours after intake of the oral fat load on nondenaturing gradient gel electrophoresis (GGE) in both patients and control subjects. Virtually no changes in the plasma concentration of HDL GGE subclasses, lipoproteins containing apolipoprotein A-I but no apolipoprotein A-II (LpA-I), or lipoproteins containing both apolipoproteins A-I and A-II (LpA-I:A-II) were induced in the postprandial state despite massive increases of large very low density lipoprotein (VLDL) and large chylomicron remnant levels (determined as apolipoproteins B-100 and B-48 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis). Strong inverse correlations with fasting plasma HDL cholesterol and the larger HDL GGE subspecies were found for large postprandial VLDL and large chylomicron remnants, whereas the corresponding relations for small VLDL and small chylomicron remnants were weaker. The relations of both large and small VLDL and chylomicron remnants to HDL cholesterol were confined to subjects in the lower fasting plasma HDL cholesterol range (< 1.2 mmol/l). None of the HDL parameters measured, either in the fasting or in the postprandial state (HDL cholesterol, HDL triglycerides, HDL GGE subclasses, LpA-I, and LpA-I:A-II), were related to the development of coronary atherosclerosis, whereas the postprandial plasma levels of small chylomicron remnants, which showed weak negative correlations with HDL, related positively to the progression of coronary atherosclerosis.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8422333 TI - Additive and synergistic effects of a low-molecular-weight, heparin-like molecule and low doses of cyclosporin in preventing arterial graft rejection in rats. AB - Arteriosclerotic intimal proliferation is one of the main long-term complications of organ transplantation. Low-molecular-weight, heparin-like molecules prevent myointimal proliferation in arterial wall injury and limit rejection in skin allografts. Cyclosporin limits rejection but has no major effect on intimal proliferation. Therefore, an experimental protocol was designed to test whether heparin-like molecules interacted with low doses of cyclosporin to prevent arterial wall immune system injury and response in a model of arterial graft rejection in normotensive and hypertensive rats. Aortic allografts were performed in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto (WKY) normotensive control rats. Four groups of 10 allografted (SHR and WKY) rats were used: one group was treated with placebo, one with low doses of cyclosporin (2 mg/kg body wt per day), one with low-molecular-weight, heparin-like molecule (1 mg/kg body wt per hour), and one with low doses of cyclosporin plus low-molecular-weight, heparin-like molecule. Ten SHRs and 10 WKYs were isografted and served as the control groups. All rats were killed 8 weeks after aortic grafting. Structural parameters of the grafted segment were measured by morphometric analysis on formalin-fixed sections with specific stains. The classical signs of immune system injury and response were present in the untreated allografts in SHRs and WKYs: inflammatory infiltration of the adventitia, medial injury, and intimal proliferative response. Low doses of cyclosporin had a significant beneficial effect on immune medial injury by increasing medial thickness and the number of remaining smooth muscle cells and decreasing the extracellular matrix injury. Cyclosporin had no protective effect on intimal proliferation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422334 TI - Osteopontin overexpression is associated with arterial smooth muscle cell proliferation in vitro. AB - To isolate the genes involved in the cell cycle G1 phase progression of arterial smooth muscle cells (SMCs), a cDNA clone (M11) was previously selected by differential hybridization screening of a mid-G1 serum-stimulated SMC cDNA library. The delay of induction after mitogenic stimulation, time of expression, and need for new protein synthesis for full expression made it possible to classify this gene in the "delayed early" gene group. Determination of the partial M11 cDNA sequence showed full homology with the osteopontin gene (secreted phosphoprotein 1, 2ar), an Arg-Gly-Asp-containing extracellular matrix protein. Osteopontin mRNA was also detected in the aorta at levels as high as in the kidney but lower than in bone, two tissues in which it has been previously detected. In vitro analysis of osteopontin expression in serum-stimulated quiescent SMCs and asynchronously cycling SMCs demonstrated that osteopontin overexpression was associated with SMC proliferation. In view of our results, the high osteopontin expression observed by others in the injured carotid artery could be explained by the involvement of SMCs in the proliferative process. Taken together, these results suggest that osteopontin may play an important role in pathological processes that are associated with arterial SMC proliferation, such as atherosclerosis or restenosis. PMID- 8422335 TI - Characterization of apoA-IV-containing lipoprotein particles isolated from human plasma and interstitial fluid. AB - Apolipoprotein (apo) A-IV has been proposed to play a role in reverse cholesterol transport. ApoA-IV-containing lipoprotein particles (A-IVLp) were isolated from human plasma and interstitial fluid and characterized by immunoaffinity chromatography. Two major A-IVLp subpopulations, lipoprotein particles containing apoA-IV with apoA-I (LpA-I:A-IV) and lipoprotein particles containing apoA-IV without apoA-I (LpA-IV), were identified. The larger subpopulation of A-IVLp is the LpA-IV that represents 70% (protein mass) of the initial particles. Only 5.8% of apoA-IV was recovered in the retained fraction after affinity chromatography with an anti-apoA-I immunosorbent. ApoA-I, apoA-II, apoA-IV, apoB, apoC-III, apoD, apoE, apoH, lecithin: cholesterol acyltransferase (LCAT), cholesteryl ester transfer (CET) protein, proline-rich protein, and a protein of Mr 59,000 were detected in the A-IVLp. These particles contain more than 20% triglycerides (lipid mass). ApoA-IV-containing particles that were isolated from plasma are heterogeneous in size, consisting of two major populations with Stokes' diameters of 10.3 nm and 9.3 nm. Both subpopulations of A-IVLp contain LCAT and CET activities and promote cholesterol efflux from cholesterol-preloaded adipose cells. These data support the hypothesis that A-IVLp particles may be involved in reverse cholesterol transport. PMID- 8422336 TI - In situ immunolocalization of lipoproteins in human arteriosclerotic tissue. AB - The concentration of serum lipoproteins, especially those of low density (LDL) and high density (HDL) lipoprotein, are related to the pathogenesis of arteriosclerosis. However, there is a lack of data concerning lipoprotein distribution in the human arteriosclerotic plaque. To detect these lipoproteins, we performed immunogold labeling on ultrathin sections of fixed and embedded human arteriosclerotic tissue. We used a panel of specific antibodies to different lipoproteins and their apolipoprotein constituents, namely LDL, formaldehyde-fixed LDL, apolipoprotein B-100, HDL, and formaldehyde-fixed apolipoprotein A-I. We also applied antibodies to alpha-actin and cathepsin D to characterize the cells and organelles involved in lipoprotein uptake and metabolism. Semiquantitative evaluation was carried out for a detailed comparison of the results obtained. Electron microscopic examination revealed that the majority of HDL and LDL in the pathological tissue was localized intracellularly in macrophage-derived foam cells and smooth muscle cells, whereas only LDL was found in the extracellular matrix. In some cases, we observed an intracellular accumulation of lipoproteins in electron-dense vesicles, which appeared to be of lysosomal origin, as shown by double labeling with an antibody to cathepsin D. These vesicles were present only in macrophage-derived foam cells, which were localized in the necrotic cores of arteriosclerotic plaques, and could not be found in healthy tissue or in the early stages of arteriosclerotic disease. PMID- 8422337 TI - Inhibition of copper-mediated oxidation of LDL by rat serosal mast cells. A novel cellular protective mechanism involving proteolysis of the substrate under oxidative stress. AB - Rat serosal mast cells, when stimulated to exocytose their cytoplasmic granules, effectively blocked the copper-mediated oxidation of low density lipoproteins (LDLs) in vitro. This effect depended on the proteolytic activity of the formed extracellular granule remnants, since specific inhibition of chymase, the neutral protease that they contain, blocked the protective effect of the mast cells. The mechanism of this chymase-mediated inhibition of LDL oxidation was found to be binding of the copper ions present in the incubation medium by peptides released from LDL on proteolytic degradation of their apolipoprotein B (apoB) component. This was verified by demonstrating that addition of such peptides to LDL--copper ion mixtures completely prevented oxidation of LDL and that this protective effect could be overcome by adding copper ions in excess. Furthermore, proteolytic degradation of the apoB of LDL, with concomitant release of copper containing peptides, left the partially degraded apoB without the copper ions necessary for propagation of LDL oxidation. These observations provide the first evidence for cell-mediated inhibition of LDL oxidation. PMID- 8422338 TI - Metabolic heterogeneity associated with high plasma triglyceride or low HDL cholesterol levels in men. AB - To further understand the factors involved in the regulation of high plasma triglyceride (TG) or low plasma high density lipoprotein cholesterol (HDL-C) levels, three groups of male subjects (normal TG with low HDL-C levels, high TG with normal HDL-C levels, and high TG with low HDL-C levels) were compared with a sample of normolipemic men with normal TG and HDL-C plasma levels. Mean age was 34 years (range, 20-42 years), and none of the subjects had plasma TG levels > 4.0 mmol/l or familial hypercholesterolemia. Both groups of subjects with high TG levels had a higher body mass index, waist circumference, waist-to-hip circumferences ratio, and a higher ratio of abdominal to femoral adipose tissue areas as measured by computed tomography when compared with normolipemic control subjects. However, during an oral glucose tolerance test only high TG-low HDL-C men had fasting hyperinsulinemia and higher plasma insulin levels compared with normolipemic subjects. In addition, the high TG-low HDL-C group showed reduced HDL apoprotein (apo) A-I levels and a low HDL2-C/HDL3-C ratio. These changes were observed along with a nonsignificant trend for a lower plasma postheparin lipoprotein lipase activity. However, among subjects with high TG and normal HDL C levels, no evidence of insulin resistance or of a reduction in postheparin lipoprotein lipase activity was observed, suggesting that the high plasma TG levels could be attributed to an increased production of apo B-containing lipoproteins, as high plasma apo B and low density lipoprotein (LDL)-apo B levels were observed in this group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422339 TI - Insulin resistance in familial and nonfamilial hypercholesterolemia. AB - High levels of very low density lipoprotein triglycerides and low levels of high density lipoprotein cholesterol have been found to be associated with insulin resistance measured by the euglycemic clamp technique. In contrast, the association of isolated hypercholesterolemia with insulin resistance has not been systematically studied. Therefore, we performed two separate studies designed to investigate the degree of insulin resistance in familial hypercholesterolemia (FH) (study 1) and nonfamilial hypercholesterolemia (non-FH) (study 2). Study 1 included eight young adults with FH and 13 corresponding control subjects. Fasting blood glucose, insulin, and C-peptide levels were similar in FH patients and control subjects during an oral glucose tolerance test. During the euglycemic hyperinsulinemic (1,200-1,300 pmol/l) clamp studies, FH patients and control subjects had similar rates of whole-body glucose uptake (73 +/- 6 versus 70 +/- 3 mumol/kg per minute, respectively; p = NS). Glucose oxidation, glucose nonoxidation, lipid oxidation, suppression of free fatty acid levels, and potassium disposal were similar in both groups. Study 2 included 25 middle-aged non-FH patients and 18 corresponding control subjects. Glucose, insulin, and C peptide responses in an oral glucose tolerance test were similar in both groups. During the euglycemic hyperglycemic clamp studies, non-FH patients and control subjects had similar rates of whole-body glucose uptake (61 +/- 3 versus 58 +/- 3 mumol/kg per minute, p = NS). In addition, glucose oxidation, glucose nonoxidation, lipid oxidation, and suppression of free fatty acid levels as well as potassium disposal were similar in non-FH patients and control subjects. We conclude that FH and non-FH are not insulin-resistant states. PMID- 8422340 TI - Isolated systolic hypertension and vessel wall thickness of the carotid artery. The Rotterdam Elderly Study. AB - We studied the association between isolated systolic hypertension (ISH) and generalized atherosclerosis as indicated by intima-media wall thickness (IMT) of the distal common carotid artery. The Rotterdam Elderly Study is a single-center study of a cohort of 11,854 elderly persons > or = 55 years old. Baseline measurements included ultrasonic evaluation of plaques and vessel wall thickness of both carotid arteries and extensive measurements of cardiovascular risk factors. Mean IMT and lumen diameter of subjects with ISH (systolic pressure > or = 160 mm Hg and diastolic pressure < 90 mm Hg) among the first 1,000 participants (n = 33) and 66 age- and sex-matched control subjects were compared. None of the subjects were using antihypertensive drug treatment, and all were free of cardiovascular disease. Mean IMT of the right common carotid artery was significantly higher in those with ISH than in normotensive subjects, with a mean difference of 0.07 mm (95% confidence interval [CI], 0.01, 0.14). Results for the left carotid artery were similar (mean difference, 0.06 mm; 95% CI, -0.01, 0.13). The end-diastolic mean lumen diameter was significantly larger in subjects with ISH than in control subjects for both right and left sides, with a mean difference of 0.70 mm (95% CI, 0.38, 1.01) and 0.48 mm (95% CI, 0.17, 0.80), respectively. Adjustment for differences in body mass index, serum lipids, smoking, and fibrinogen did not materially change the findings. Furthermore, atherosclerotic plaques were more frequently observed among those with ISH compared with control subjects, with a mean difference of 12% (95% CI, -1, 25).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422341 TI - Arterial intimal hyperplasia after occlusion of the adventitial vasa vasorum in the pig. AB - Oxygenation of the arterial wall is provided by diffusion of oxygen outward from the main vessel lumen and inward from the adventitial vasa vasorum. In a group of four Yucatan miniature pigs the oxygenation profiles across the superficial femoral arteries were recorded by polarographic oxygen microelectrodes. The profiles obtained suggested a relatively poorly oxygenated media (a trough value of approximately 25% that of the intimal oxygenation) with a progressive rise in oxygenation toward the intimal and adventitial surfaces. In four other survival experiments, occlusion of the adventitial vasa vasorum by flush ligation of the arterial branches that supply them resulted in the production of a focal, intimal hyperplastic lesion that was absent in control vessels (intimal to medial ratios [mean +/- SEM] of 0.053 +/- 0.008, n = 8, p < 0.001 and 0.013 +/- 0.001, n = 8, respectively). By electron microscopy this lesion was seen to be composed mainly of smooth muscle cells. This evidence would support the hypothesis that arterial wall hypoxia may be involved in the initiation of intimal hyperplasia. It is proposed that human atherosclerosis may be initiated by occlusion of the vasa vasorum and concomitant hypoxia. PMID- 8422342 TI - Potential of 99mTc-LDLs labeled by two different methods for scintigraphic detection of experimental atherosclerosis in rabbits. AB - In this study we evaluated two different 99mTc-labeling techniques to produce 99mTc-low density lipoprotein (99mTc-LDL) suitable for the scintigraphic delineation of experimental atherosclerotic lesions. The two methods are 1) a procedure that uses stannous chloride and sodium borohydride (borohydride method) and 2) a procedure that uses sodium dithionite as a reducing agent and that has been successfully applied in previous scintigraphic atherosclerosis detection (dithionite method). 99mTc-LDL produced by either method was injected into New Zealand White rabbits with diet-induced atherosclerotic plaques and in control rabbits. Scintigraphic images were taken 10 minutes (t = 0) and 1, 4, 8, 16, and 24 hours after injection. Clearance of plasma radioactivity was also studied. Stability of the 99mTc-LDL complex in the circulation was examined by size exclusion chromatography of plasma samples. After scintigraphy, the animals were killed, and the biodistribution of radioactivity was determined. The thoracic and abdominal aortas appeared in scintigraphic images to accumulate 99mTc over their entire length with either 99mTc-LDL preparation. The sparse imaging of focal atherosclerosis was found to be due to the fact that the aortas were covered with confluent atherosclerotic lesions. Scintigraphic image analysis showed that 24 hours after injection, the accumulated radioactivity in the abdominal aorta of the atherosclerotic rabbits was 57% and 54%, respectively, of the accumulated radioactivity in the abdominal aorta at t = 0 when the borohydride versus the dithionite method was used. In the control animals this value was 25% for the dithionite method, whereas in the borohydride method the aortas could not be detected in the images at t = 24 hours.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422343 TI - The effects of olive oil, hydrogenated palm oil, and omega-3 fatty acid-enriched diets on megakaryocytes and platelets. AB - Unsaturated fatty acids are thought to prevent thrombotic and arteriosclerotic disease, whereas saturated fatty acids are thought to increase the incidence of these disorders. However, the effects of these diets on megakaryocytes and platelets are not well understood. We compared the effects of diets enriched with 8.4% olive oil, 8.4% hydrogenated palm oil, or 10.2% omega-3 fatty acid ethyl esters on guinea pig megakaryocytes and platelets. In plasma, changes in fatty acid composition reflected the composition of each diet. However, in platelets and megakaryocytes, hydrogenated palm oil induced a decrease in 16:0 and an increase in 18:2 while the olive oil diet caused a marked increase in 18:1 and a decrease in most other fatty acids. The differences in the effects of the diets on cellular versus plasma fatty acids suggest that megakaryocytes and platelets have an extensive capacity to regulate their fatty acid composition. Thrombocytosis occurred with the omega-3 fatty acid-enriched diet: 12.9 +/- 1.78 x 10(5) compared with 7.45 +/- 1.08 x 10(5) platelets per microliter of platelet rich plasma in control animals. There was an increase in megakaryocyte size, ploidy, and morphological stage (cytoplasmic maturation) with the omega-3 fatty acid-enriched diet but not with the other diets. The omega-3 fatty acid-enriched diet decreased platelet thromboxane production while the other diets had no effect. Platelet hypersensitivity was suggested in collagen aggregation studies with olive oil but not with the hydrogenated palm oil diet. Although saturated fatty acid diets are thought to be atherogenic, this diet had no affect on platelet function.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422344 TI - Arterial alterations with aging and high blood pressure. A noninvasive study of carotid and femoral arteries. AB - Noninvasive in situ evaluations of pulsatile changes of blood pressure and arterial diameter were performed at the sites of the common carotid and femoral arteries in a population of 78 untreated normotensive and hypertensive subjects. Arterial segments were studied by using an original echo-tracking technique for internal diameter and validated applanation tonometry for local pulse pressure measurements. Whereas mean arterial pressure is known to be identical in all parts of the arterial tree, pulse pressure was significantly lower in the carotid (52.7 +/- 2.2 mm Hg) than in the brachial (62.0 +/- 2.0 mm Hg) or femoral (62.5 +/- 2.5 mm Hg) arteries. Despite a higher pulse pressure and diastolic diameter, the femoral artery had a lower pulsatile change in diameter (3.47 +/- 0.18% versus 6.07 +/- 0.28%; p < 0.0001) and distensibility coefficient (9.36 +/- 0.58 versus 21.60 +/- 1.75 x 10(-3) kPa-1) than the carotid artery. Local cross sectional compliance of the carotid artery was higher than that of the femoral artery (7.42 +/- 0.46 versus 6.20 +/- 0.28 m2.kPa-1.10(-7); p < 0.05). Whereas age was strongly correlated with arterial parameters at the site of the carotid artery (pulse pressure: r = 0.54, p < 0.0001; pulsatile change in arterial diameter: r = -0.62, p < 0.0001; distensibility coefficient: r = -0.70, p < 0.0001), no significant correlation was observed at the femoral artery. Mean blood pressure was the second factor of carotid artery alterations: the higher the mean blood pressure, the lower the distensibility of this artery (r = -0.36, p < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422345 TI - Inhibition of exercise-induced shortening of bleeding time by fish oil in familial hypercholesterolemia (type IIa). AB - Fourteen patients suffering from familial hypercholesterolemia (type IIa) participated in a double-blind, placebo-controlled trial that evaluated the effects of fish oil ethyl ester (K-85, 5.7 g/day) or a hydroxymethylglutaryl coenzyme A reductase inhibitor (lovastatin, 40 mg/day) alone or in combination on lipid metabolism and bleeding time at rest and after standardized exercise. Lovastatin treatment reduced total cholesterol (-27%), low density lipoprotein cholesterol (-37%), and triglycerides (-18%), whereas high density lipoprotein cholesterol increased significantly (14%). K-85 affected total (-4%), low density lipoprotein (-9%), and high density lipoprotein (+7%) cholesterol insignificantly, whereas the triglyceride level decreased by 24% (p < 0.001). The combined regimen caused an additive decrease in the triglyceride level (41%), which differed significantly (p < 0.01) from that gained by lovastatin alone. Under basal conditions the bleeding time was not influenced by the different interventions. Standardized exercise shortened the bleeding time by 19% (p < 0.001) and 16% (p < 0.001) before intervention and after lovastatin treatment, respectively. After K-85 alone or in combination with lovastatin, the exercise induced shortening of the bleeding time was totally inhibited, which may reflect a favorable influence of fish oil on the platelet-vessel wall interaction in these high-risk patients. PMID- 8422346 TI - Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment. PMID- 8422347 TI - Selective isotopic enrichment of synthetic RNA: application to the HIV-1 TAR element. AB - The introduction of isotopically enriched nucleotides into NMR quantities of a synthetic 29-mer RNA derived from the HIV-1 TAR element is described. RNA enriched in 13C and/or 15N is produced by a procedure which involves isolation of whole cellular RNA from Escherichia coli, nucleolysis, separation of mononucleotides, chemical or enzymatic pyrophosphorylation, and in vitro transcription by T7 RNA polymerase. Spectral characteristics of each residue type are examined in isolation. 13C chemical shifts provide an alternative method to determine ribose puckers for larger RNAs. Nonprotonated sites such as purine N7 groups can now be monitored through the use of multiple-bond 1H-15N coupling. When applied conservatively, coordinate analysis of chemical shift values should prove valuable for NMR studies of RNA structure and recognition. 1H, 13C, and 15N chemical shift data suggest that TAR residue A35 has an unusual local environment, consistent with extrusion of its base from the terminal loop. PMID- 8422348 TI - Acetylcholinesterase: electrostatic steering increases the rate of ligand binding. AB - Brownian dynamics simulations have been used to calculate the diffusion controlled rate constants for the binding of a positively charged ligand to several models of acetylcholinesterase (AChE). The crystal structure was used to define the detailed topography and the active sites of the dimeric enzyme. The electric field around AChE was then computed by solving the Poisson equation for different charge distributions in the enzyme at zero ionic strength. These fields were used in turn to calculate the forces on the diffusing ligand. Significant increases in the rate constant resulted in going from a model with no charges to one with the net charges concentrated at the centers of the monomers and then to a model with a realistic distribution of charges throughout the enzyme. The results show that electrostatic steering of ligands contributes to the high rate constants that are observed experimentally for AChE. PMID- 8422349 TI - Mechanism of bacterial bioluminescence: 4a,5-dihydroflavin analogs as models for luciferase hydroperoxide intermediates and the effect of substituents at the 8 position of flavin on luciferase kinetics. AB - Bioluminescence catalyzed by bacterial luciferases was measured using FMN, iso FMN (6-methyl-8-nor-FMN), and FMN analogs carrying the following substituents at position 8: -H, -Cl, -F, SMe, SOMe, -SO2Me, or -OMe. The first-order rate constants for the decay of light emission correlate with the one-electron oxidation potentials of the 4a,5-dihydro forms of the FMN analogs. To determine the values of these potentials, isoalloxazine (flavin) derivatives having the 4a,5-propano-4a,5-dihydro structure and -H, -CH3, -Cl, -OCH3, and -NH2 as substituents at position 8 have been synthesized as models for the 4a-peroxy-4a,5 dihydroflavin intermediates occurring during catalysis by the flavin-dependent monooxygenase luciferase. The tetrahydropyrrole ring between positions 4a and 5 of these isoalloxazine derivatives stabilizes the 4a,5-dihydroflavin by impeding formation of the thermodynamically more stable 1,5-dihydro form. One-electron oxidation potentials (Eobs) were measured by cyclic voltammetry and used to determine the empirical coefficients in the Swain equation. On the basis of this, the one-electron oxidation potentials of 4a,5-propano-4a,5-dihydro analogs with other substituents in position 8 were calculated (Ecalc). The bioluminescence reaction rate is fastest with FMN analogs of lowest oxidation potential; i.e., the slope of the correlation is negative. This indicates that in the rate limiting step the 4a,5-dihydroflavin moiety donates negative charge. The results are compatible with an intramolecular, chemically initiated electron exchange luminescence mechanism for the bacterial luciferase reaction.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422350 TI - Statistical clustering techniques for the analysis of long molecular dynamics trajectories: analysis of 2.2-ns trajectories of YPGDV. AB - The microscopic interactions and mechanisms leading to nascent protein folding events are generally unknown. While such short time-scale events are difficult to study experimentally, molecular dynamics simulations of peptides can provide a useful model for studying events related to protein folding initiation. Recently, two extremely long molecular dynamics simulations (2.2 ns each) were carried out on the pentapeptide Tyr-Pro-Gly-Asp-Val [Tobias, D. J., Mertz, J. E., & Brooks, C. L., III (1991) Biochemistry 30, 6054-6058] that forms stable reverse turns in solution. Tobias et al. examined folding events in this large system (approximately 30,000 conformations) using traditional methods of trajectory analysis. The shear magnitude of this problem prompted us to develop an automated approach, based on self-organizing neural nets, to extract the key features of the molecular dynamics trajectory. The neural net is used to perform conformational clustering, which reduces the complexity of a system while minimizing the loss of information. The conformations were grouped together using distances in dihedral angle space as a measure of conformational similarity. The resulting clusters represent "conformational states", and transitions between these states were examined to identify mechanisms of conformational change. Many conformational changes involved the rotation of only a single dihedral angle, but concerted angle changes were also found. Most of the conformational information in the 30,000 samples from the full trajectories was retained in the relatively few resultant clusters, providing a powerful tool for analysis of an expanding base of large molecular simulations. PMID- 8422351 TI - Capping interactions in isolated alpha helices: position-dependent substitution effects and structure of a serine-capped peptide helix. AB - The influence of an amino acid on the stability of alpha-helical structure depends on the position of the residue in the helix with respect to the ends. Short alpha helices in proteins are stabilized both by H-bonding of the main chain NH and CO groups and by capping interactions between side chains and unfulfilled peptide groups at the N and C termini. Peptide models based on consensus position-dependent helix sequences allow one to model capping effects in isolated helices and to establish a base line for these interactions in proteins. We report here an extended series of substitutions in the cap positions of our peptide models and the solution structure of peptide S3, with serine at the N-cap position defined as the N-terminal residue with partly helix and partly coil conformation. The resulting model, determined by 2D 1H NMR, is consistent with a structure at the N-cap involving H-bonding between the serine gamma oxygen and the peptide NH of the glutamic acid residue three amino acids toward the C terminus. A bifurcated H-bond of Ser O gamma with the NH of Asp5 is possible also, since this group is within interacting distance. This provides direct evidence that specific side-chain interactions with the main chain stabilize isolated alpha-helical structure. PMID- 8422352 TI - Comparison of backbone and tryptophan side-chain dynamics of reduced and oxidized Escherichia coli thioredoxin using 15N NMR relaxation measurements. AB - The backbone and tryptophan side-chain dynamics of both the reduced and oxidized forms of uniformly 15N-labeled Escherichia coli thioredoxin have been characterized using inverse-detected two-dimensional 1H-15N NMR spectroscopy. Longitudinal (T1) and transverse (T2) 15N relaxation time constants and steady state (1H)-15N NOEs were measured for more than 90% of the protonated backbone nitrogen atoms and for the protonated indole nitrogen atoms of the two tryptophan residues. These data were analyzed by using a model free dynamics formalism to determine the generalized order parameter (S2), the effective correlation time for internal motions (tau e), and 15N exchange broadening contributions (Rex) for each residue, as well as the overall molecular rotational correlation time (tau m). The reduced and oxidized forms exhibit almost identical dynamic behavior on the picosecond to nanosecond time scale. The W31 side chain is significantly more mobile than the W28 side chain, consistent with the positions of W31 on the protein surface and W28 buried in the hydrophobic core. Backbone regions which are significantly more mobile than the average include the N-terminus, which is constrained in the crystal structure of oxidized thioredoxin by specific contacts with a Cu2+ ion, the C-terminus, residues 20-22, which constitute a linker region between the first alpha-helix and the second beta-strand, and residues 73-75 and 93-94, which are located adjacent to the active site. In contrast, on the microsecond to millisecond time scale, reduced thioredoxin exhibits considerable dynamic mobility in the residue 73-75 region, while oxidized thioredoxin exhibits no significant mobility in this region. The possible functional implications of the dynamics results are discussed. PMID- 8422353 TI - Theoretical predictions of DNA hairpin loop conformations: correlations with thermodynamic and spectroscopic data. AB - A computational procedure for generating conformations of DNA hairpin loop structures from a broad range of low-energy starting states is described. The starting point of the modeling is the distribution of oligonucleotide chain conformations obtained from Monte Carlo simulations of feasible dinucleotide steps. Structures which meet the spatial criteria for hairpin loop formation are selected from the distributions and subsequently minimized using all-atom molecular mechanics. Both d(CTnG) and d(CAnG) oligomers, where n = 3, 4, or 5, are modeled. These sequences are chosen because of the large number of published NMR and thermodynamic studies on DNA hairpins containing thymine or adenine residues. The minimized three-dimensional hairpin loop structures are compared with one another as well as analyzed in terms of available experimental data. The computational approach provides the first detailed analysis of DNA hairpin loop structure in terms of a multistate conformational model. Investigation of the minimized conformations reveals several interesting structural features. First, hairpin loops of the same sequence adopt several distinctly different conformations, as opposed to minor variants of the same equilibrium structure, that could potentially interconvert in solution. Second, in contrast to double helical nucleic acids, the hairpin loop models exhibit hydrophobic and hydrophilic surfaces. The different disposition of hydrophobic groups in loops versus duplexes could modulate both protein-nucleic acid interactions and nucleic acid self-associations. Third, perpendicular aromatic interactions of loop residues are observed in many of the computed hairpins. This sort of interaction might be important in the stabilization of non-hydrogen-bonded nucleic acid secondary and tertiary structures. The predicted structural features in the models help, in addition, to account for the unusual thermodynamic properties of DNA hairpin loops. Comparison of the theoretically-generated NOEs in different structures further reveals that very different molecular structures and interactions can, in principle, produce the same NOEs. The multistate description suggested by this observation differs from the conventional interpretation of DNA solution structure in terms of the fluctuations about a single preferred chain conformation. There is not necessarily only one set of closely related structures consistent with the observed data. PMID- 8422354 TI - Symmetry and molecular structure of a DNA triple helix: d(T)n.d(A)n.d(T)n. AB - A structure for the triple helix d(T)n.d(A)n.d(T)n consistent with recent infrared spectral data is proposed, and its salient features are discussed. The present structure preserves the pseudodyad between the Watson-Crick base-paired adenine and thymine strands and in addition has a pseudorotational symmetry relating the Hoogsteen-paired adenine and thymine strands. The simultaneous presence of these two symmetries gives rise to a dyad between the two thymine polynucleotides. These symmetries result in identical backbone conformations for all three strands, unlike any previously proposed model for a triple helix. The proposed structure has an axial rise per residue of 3.26 A and 12 residues per turn obtained from X-ray fiber diffraction [Arnott S., & Selsing, E. (1974) J. Mol. Biol. 88, 509-521]. The present structure is structurally and conformationally similar to double helical B-form DNA and has sugar pucker in the C2'-endo region. This structure is fundamentally different from the one proposed by Arnott and co-workers, which was based on structural and conformational features similar to double helical A-form DNA with C3'-endo sugar pucker. It is stereochemically satisfactory, and it does not have the disallowed nonbonded distances present in the earlier model of Arnott and co-workers. It is energetically much more favorable than their structure. Coordinates of the present structure are given. PMID- 8422355 TI - The sugars in chromomycin A3 stabilize the Mg(2+)-dimer complex. AB - Chromomycin A3 (CRA3) is a glycosylated antitumor antibiotic that binds as a dimer to the minor groove of DNA, with a Mg2+ cation (or another divalent cation with a radius less than 0.85 A) forming the center of the dimer. It has been shown that the chromose sugars are necessary for DNA binding [Kaziro & Kamiyama (1967) J. Biochem. (Tokyo) 62, 424-429; Kamiyama (1968) J. Biochem. (Tokyo) 63, 566-572], although the reason for this has not been addressed. We have investigated the role that the chromose sugars play in metal complexation in solution (methanol) by comparing the optical behavior of CRA3 and its aglycon, CRN, in the presence of various divalent metals (Mg2+, Ni2+, and Ca2+). The results show that CRA3 forms a dimeric complex [i.e., (CRA3)2M, where M is a metal ion] in the presence of 1 mol equiv of either Ni2+ or Mg2+ but a 1:1 complex in the presence of the much larger Ca2+. In contrast, CRN forms a 1:1 complex (CRN.M)+ with all three metals under identical conditions (1:1 mole ratio of drug to metal). Thus, for the smaller metal ions the sugars stabilize the 2:1 CRA3-metal complex in solution. NMR data on the 2:1 CRA3-Mg2+ complex show that the trisaccharide of one CRA3 molecule lies in close proximity to the chromophore of the other CRA3 molecule. This interaction, which is also present in the Mg(2+) CRA3-DNA complex [Gao & Patel (1989) Biochemistry 28, 751-762], appears to be related to the stability of the dimer in solution.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422356 TI - In vivo evidence that UV-induced C-->T mutations at dipyrimidine sites could result from the replicative bypass of cis-syn cyclobutane dimers or their deamination products. AB - The major mutations induced by UV light are C-->T transitions at dipyrimidines and arise from the incorporation of A opposite the C of dipyrimidine photoproducts. The incorporation of A has most often been explained by the known preference of a polymerase to do so opposite noninstructional DNA damage such as an abasic site (A rule). There are also mechanisms that suppose, however, that cis-syn dipyrimidine photodimers are instructional. In one such mechanism (tautomer bypass), the incorporation of A is directed by the tautomer of a C of a dimer that is equivalent in base-pairing properties to U [Person et al. (1974) Genetics 78, 1035-1049]. In another mechanism (deamination bypass), the incorporation of A is directed by a U of a dimer that results from the deamination of the C of a dimer [Taylor & O'Day (1990) Biochemistry 29, 1624 1632]. The viability of these mechanisms was tested by obtaining the mutation spectrum of a TU dimer in Escherichia coli by application of a standard method for site-directed mutagenesis. To this end, a 41-mer containing a site-specific TU dimer was constructed via ligation of a dimer-containing decamer that was produced by triplet-sensitized irradiation and used to prime DNA synthesis on a uracil-containing (+) strand of an M13 clone containing a double mismatch opposite the dimer. The reaction mixture was used to transfect a uracil glycosylase proficient, photoproduct repair deficient E. coli host, and all progeny phage weakly hybridizing to the parental (+) or (-) strands were sequenced. Under non-SOS conditions the TU dimer almost completely blocked replication, while under SOS conditions it directed the incorporation of two As with much higher specificity (96%) than would an abasic site. The implications of these results to the mechanism of the UV-induced TC-->TT mutation, and by extension to the CT-->TT, CC-->TC, CC-->CT, and the tandem CC-->TT mutations, are discussed. PMID- 8422357 TI - Assignment of the inter- and intramolecular disulfide linkages in recombinant human macrophage colony stimulating factor using fast atom bombardment mass spectrometry. AB - The disulfide bridges in recombinant human macrophage colony stimulating factor (rhM-CSF), a 49-kDa homodimeric protein, were assigned. The 18 cysteines in the dimer form three intermolecular and two sets of three intramolecular disulfide bonds. The intermolecular disulfide bridges hold the dimer together and form symmetric bonds in which Cys31 and Cys157/Cys159 from one monomer unit are linked to the corresponding cysteines of the second monomer. The intramolecular disulfide bonds are located between Cys7-Cys90, Cys48-Cys139, and Cys102-Cys146, respectively. The resistance of native M-CSF to proteolytic cleavage was overcome by an initial chemical cleavage reaction using BrCN. The close proximity of four cysteines (Cys139, Cys146, Cys157, and Cys159) results in a tight core complex that makes the protein undigestable for most proteases. Digestion using endoprotease Asp-N resulted in cleavage at Asp156 near the C-terminal end of this region, thereby opening the complex structure. PMID- 8422358 TI - Effects of plasmid length and positioned nucleosomes on chromatin assembly in vitro. AB - Histone H5 induces extensive nucleosome alignment in vitro, with a 210 +/- 5 base pair (bp) average unit repeat, on some of the constructs derived from plasmid pBR327. Plasmid pBR327 itself aligns nucleosomes poorly, even though it possesses a chromatin organizing region which nucleates the alignment reaction [Jeong et al. (1991) J. Mol. Biol. 222, 1131-1147]. Examination of various regions of pBR327 chromatin by Southern hybridization revealed no substantial regional differences, suggesting an essentially all-or-none alignment mechanism. Twenty four pBR327 deletion constructs, with the chromatin organizing region intact, were analyzed for nucleosome alignment in vitro, in addition to the six previously described. Although nucleosome alignment on plasmids of size greater than 5 kb was not affected by small length changes, circular plasmids with total lengths between 2400 and 3600 bp generally permitted alignment only when their lengths were close to integer multiples of 210 +/- 3 bp. The measured repeat lengths for the large plasmids and the smaller ones that aligned nucleosomes were all 210 bp, within experimental precision. The failure of two approximately 3.2 kb plasmids to align nucleosomes, even though their lengths were close to 15 x 210 bp, could be attributed to the effects of four strongly positioned nucleosomes that form on pBR327 sequences. Evidence is provided that nucleosome arrays can be quasicrystalline and are capable of transmitting information over a distance of more than 2 kb. PMID- 8422359 TI - Effect of the Z mutation on the physical and inhibitory properties of alpha 1 antitrypsin. AB - A major feature of the structure of alpha 1-antitrypsin is a five-stranded A sheet into which the reactive center loop inserts after cleavage. We describe here the effect of the Z mutation (342Glu to Lys) at the head of the fifth strand of the A-sheet on the mobility of the reactive center loop and hence on the physical properties of the antitrypsin molecule. The mutant Z but not the normal M antitrypsin spontaneously polymerizes at 37 degrees C by a mechanism involving the insertion of the reactive center loop of one molecule into the A-sheet of a second. It is demonstrated that Z antitrypsin polymerized after incubation with 1.0 M guanidinium chloride at 37 degrees C at the same rate as M antitrypsin. Reducing the temperature to 4 degrees C favored the formation of the L-state in M antitrypsin in which the loop is stably incorporated into the A-sheet, but resulted in loop-sheet polymerization in Z antitrypsin. Z, like M antitrypsin, undergoes the S to R transition, but we show that the accompanying change in thermal stability results from loop-sheet polymerization (S) which can be prevented by the insertion of the cleaved strand of the reactive center loop into the A-sheet (R). Z antitrypsin has a reduced association rate constant with neutrophil elastase [(5.3 +/- 0.06) x 10(7) and (1.2 +/- 0.02) x 10(7) M-1 s-1 for M and Z, respectively], but both M and Z antitrypsin had Ki values of less than 5 pM.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422360 TI - Opossum serum alpha 1-proteinase inhibitor: purification, linear sequence, and resistance to inactivation by rattlesnake venom metalloproteinases. AB - Opossum (Didelphis virginiana) serum was fractionated with (NH4)2SO4 and then chromatographed on DEAE-Sepharose and phenyl-Sepharose. Affinity chromatography on a protein A-Sepharose-antibody column removed traces of opossum serum metalloproteinase inhibitors, and resulted in a homogeneous preparation of opossum alpha 1-proteinase inhibitor (alpha 1-PI). The inhibitor is a single chain glycoprotein (17.7% carbohydrate) with an estimated M(r) = 54,000. An opossum liver cDNA library was immunoscreened, and clones containing cDNA encoding for the open reading frame for opossum alpha 1-PI were isolated. The cDNA inserts contained nucleotide sequences corresponding to the amino-terminal and an internal peptide sequence of opossum alpha 1-PI which had been separately determined by protein sequence analysis. The entire inserts coded for a protein consisting of a 21-residue signal peptide and a 389-residue mature protein. Opossum alpha 1-PI shows 51-58% identity with other mammalian alpha 1-PI amino acid sequences, and the conserved residues expected for a member for the serpin family have been retained. The carbohydrate attachment sites and the reactive site residues (M-S) of opossum alpha 1-PI are identical to those of human alpha 1 PI. Opossum alpha 1-PI formed stable enzyme/inhibitor complexes with trypsin, chymotrypsin, and human neutrophil elastase, but did not react with thrombin or with snake venom serine proteinases. Opossum alpha 1-PI was inactivated by papain or Pseudomonas aeruginosa elastase, and electrophoretic analysis of the reaction products indicated limited proteolysis in the reactive site loop of the inhibitor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422361 TI - Differential scanning calorimetric study of the effect of cholesterol on the thermotropic phase behavior of a homologous series of linear saturated phosphatidylcholines. AB - We have studied the effects of cholesterol on the thermotropic phase behavior of aqueous dispersions of a homologous series of linear saturated phosphatidylcholines, using high-sensitivity differential scanning calorimetry and an experimental protocol which ensures that broad, low-enthalpy phase transitions are accurately monitored. We find that the incorporation of small amounts of cholesterol progressively decreases the temperature and the enthalpy, but not the cooperativity, of the pretransition of all phosphatidylcholines exhibiting such a pretransition and that the pretransition is completely abolished at cholesterol concentrations above 5 mol % in all cases. The incorporation of increasing quantities of cholesterol also alters the main or chain-melting phase transition of these phospholipid bilayers in both hydrocarbon chain length-dependent and hydrocarbon chain length-independent ways. At cholesterol concentrations of from 1 to 20-25 mol %, the DSC endotherms of all phosphatidylcholines studied consist of a superimposed sharp and broad component, the former ascribed to the melting of cholesterol-poor and the latter to the melting of the cholesterol-rich phosphatidylcholine domains. The temperature and cooperativity of the sharp component are reduced only slightly and in a chain length-independent manner with increasing cholesterol concentration, an effect we ascribe to the colligative effect of the presence of small quantities of cholesterol at the domain boundaries. Moreover, the enthalpy of the sharp component decreases and becomes zero at 20-25 mol % cholesterol for all of the phosphatidylcholines examined.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422362 TI - Membrane insertion and antibody recognition of a glycosylphosphatidylinositol anchored protein: an optical study. AB - The kinetics of binding of a glycolipid-anchored protein (the promastigote surface protease, PSP) to planar lecithin bilayers is studied by an integrated optics technique, in which the bilayer membrane is supported on an optical wave guide and the phase velocities of guided light modes in the wave guide are measured. From these velocities, the optical parameters of the membrane and PSP layers deposited on the waveguide are determined, yielding in particular the mass of PSP bound to the membrane, which is followed in real time. From a comparison of the binding rates of PSP and PSP from which the lipid moiety has been removed, it is shown that the lipid moiety plays a key role in anchoring the protein to the membrane. Specific and nonspecific binding of antibodies to membrane-anchored PSP is also investigated. As little as a fifth of a monolayer of PSP is sufficient to suppress the appreciable nonspecific binding of antibodies to the membrane. PMID- 8422363 TI - Membrane partition of fatty acids and inhibition of T cell function. AB - Short-term exposure to elevated levels of free fatty acids (FFA) perturbs a variety of cellular functions. It is frequently observed that cis-unsaturated fatty acids (FA) mediate these perturbations while trans-unsaturated or saturated FA are relatively inert. This dichotomy has generally been ascribed to the differential effects of these FA on membrane structure, specifically, that cis but not saturated FA alter lipid acyl chain order. Direct support for this view, however, is lacking because membrane partition of FA has not been determined for the relevant FA and for the conditions of the functional studies. Previous measurements of membrane partition of natural long-chain FA have relied on the determination of the amount bound to the membrane rather than the aqueous-phase concentration of FA. Because [FFA] is low, however, the partition coefficient (Kp) is relatively insensitive to the membrane-bound concentration and, therefore, accurate determinations of Kp require direct [FFA] measurements. In this study FA partition between the aqueous phase and either lipid bilayer vesicles or cytotoxic T lymphocytes (CTL) was measured using a recently developed fluorescent indicator of FFA. This indicator is composed of recombinant intestinal fatty acid binding protein derivatized with acrylodan and is termed ADIFAB. Using ADIFAB, partition coefficients were determined for seven saturated and cis-unsaturated FA under conditions that parallel those in which cis FA have been shown to inhibit CTL signaling. In general, Kp values were approximately an order of magnitude greater than previous determinations and were found to be greater for the noninhibitory saturated FA than the inhibitory cis FA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422364 TI - Subunit conformational changes accompanying bacteriophage P22 capsid maturation. AB - In double-stranded DNA bacteriophages, packaging of dsDNA requires the transformation of a precursor procapsid into a mature viral capsid. Lattice expansion and release of scaffolding subunits accompanying DNA packaging. Three dimensional structures of procapsid and mature phage lattices demonstrate that the capsid transformation involves substantial changes in subunit environment. Since this transformation occurs without subunit dissociation, it represents a transition between at least two stable subunit conformations. Using Raman spectroscopy, we have identified changes in coat protein secondary structure and side-chain environments which accompany the capsid transformation. The subunits of procapsid shells contain only 2.0 +/- 0.4% more alpha-helix and less beta sheet than those of mature capsids; however, numerous side chains are substantially altered by the transformation, including tyrosines, tryptophans, phenylalanines, and aliphatics, which are widely distributed through the subunit sequence. We propose, therefore, that procapsid expansion is accomplished through the relative motion of coat subunit domains with little change in secondary structure. Such hinge-bending conformational transitions may couple ATP-dependent dsDNA condensation with shell expansion. PMID- 8422365 TI - Potential role of two novel elastase-like enzymes in processing pro-transforming growth factor-alpha. AB - Transforming growth factor-alpha (TGF-alpha) is a mitogenic peptide produced by tumor cells and by virally and chemically transformed cells in culture. TGF-alpha is almost certainly derived from its precursor protein (pro-TGF-alpha) by limited proteolysis, but the physiologically relevant processing enzyme(s) is(are) unknown. We now report that oncogenically transformed rat liver epithelial cells (known to secrete TGF-alpha) and Schwann cells in culture transfected with SV40 T antigen (which are now reported to express mRNA encoding pro-TGF-alpha) contain membrane associated, neutral pH, serine proteinases which are elastase-like in their substrate specificity, but elastase is not known to be associated with these cell types. In both cell types, the enzyme is associated with a subcellular fraction enriched for microsomes and plasma membranes. Furthermore, the enzyme appears to be specifically induced 4-fold in the transformed epithelial cells as compared with the level of enzyme present in the nontransformed parental cells. The enzymes have been purified approximately 20,000-fold to near homogeneity (50 60 units/mg) and are virtually identical with regard to their molecular weights (38,000) and other physiochemical properties. Results obtained with numerous synthetic peptide substrates show the enzymes prefer nonpolar residues such as Ala and Val in the P1 and P2 positions, but promiscuity of cleavage specificity observed with long-chain peptide substrates is attributed to the absence of structure in these peptides. Thus, although these enzymes may be involved in processing pro-TGF-alpha at the plasma membrane of the cell, it is just as likely that these enzymes play other physiological roles in the parental and/or transformed cells and that there is no specific endoproteolytic processing enzyme of pro-TGF-alpha. PMID- 8422366 TI - Effects of temperature on ADP-ribosylation factor stimulation of cholera toxin activity. AB - The effects of cholera toxin, a secretory product of Vibrio cholerae, result from ADP-ribosylation of the stimulatory guanine nucleotide-binding (Gs) protein of the adenylyl cyclase system. Cholera toxin A subunit (CTA) also uses agmatine, a simple guanidino compound, several proteins unrelated to Gs, and CTA itself as alternative ADP-ribose acceptors. The effects of toxin occur in the jejunum presumably at body core temperature. With agmatine as a model substrate, the optimal temperature for CTA-catalyzed ADP-ribosylation was 25-30 degrees C, and that for CTA-catalyzed auto-ADP-ribosylation was 20-25 degrees C. Both activities were significantly less at 37 degrees C, reflecting lower initial velocities, not heat-inactivation of the toxin. All the transferase activities of CTA are enhanced by ADP-ribosylation factors (ARFs), approximately 20-kDa guanine nucleotide-binding proteins that are ubiquitous in mammalian cells. Phospholipids and a soluble brain ARF, in a GTP-dependent manner, activated toxin NAD:agmatine ADP-ribosyltransferase activity; their simultaneous effect was maximal at physiological temperatures (approximately 37 degrees C). At lower temperatures, the stimulation by ARF was much less. There were similar effects on other toxin catalyzed reactions, notably, the ADP-ribosylation of Gs alpha and the hydrolysis of NAD. Thus, host factors, such as ARF and phospholipid, synergistically increase cholera toxin activity at 37 degrees C and may be important in toxin action in the mammalian gut. PMID- 8422367 TI - Expression of the mature and the pro-form of human sterol carrier protein 2 in Escherichia coli alters bacterial lipids. AB - Sterol carrier protein 2 (SCP2) is a protein that is believed to be involved in the intracellular transport of cholesterol and phospholipids. Expression in mammalian COS cells of a cDNA encoding SCP2 revealed that the mature protein is synthesized as a pro-form containing a 20-residue amino-terminal leader sequence. The function of this presequence is currently not known, and pro-SCP2 is generally not detected in tissues. In order to obtain large quantities of pro SCP2 as well as the mature form of human SCP2, Escherichia coli expression plasmids were constructed. Both proteins were produced in high yield (10-30% of the total cell protein) and were found in the supernatant fraction after cell lysis. Recombinant human SCP2 and pro-SCP2 were purified to homogeneity by acid precipitation followed by ion-exchange chromatography. Both recombinant human SCP2 and pro-SCP2 had sterol exchange activity similar to that seen with SCP2 purified from rat liver. In addition, the lipid content of SCP2- and pro-SCP2 producing E. coli was analyzed. Acidic lipids were significantly increased in the transfected cells. Specifically, fatty acids were increased 2-3-fold, phosphatidylglycerol was increased 2-fold, and lipid A was increased 3-4-fold, while neutral lipids were decreased 2-3-fold as compared to control cells. This alteration of the lipid composition of E. coli expressing SCP2 or pro-SCP2 is consistent with the proposed role for SCP2 in intracellular lipid movement. PMID- 8422368 TI - Human nonpancreatic secreted phospholipase A2: interfacial parameters, substrate specificities, and competitive inhibitors. AB - The rate and equilibrium parameters for the interfacial catalysis by recombinant human nonpancreatic secreted phospholipase A2 were determined. Results show that the enzyme binds to anionic interfaces with considerably higher affinity than to zwitterionic interfaces. The extent of hydrolysis per enzyme on anionic vesicles in the processive scooting mode shows that the enzyme is fully catalytically active as a monomer. Among several secreted phospholipases A2 tested, the human nonpancreatic secreted enzyme is unique in its ability to undergo slow intervesicle exchange either by dissociation from the interface followed by binding to a different vesicle or by promoting the fusion of vesicles. The equilibrium dissociation constants for calcium, substrate analogs, reaction products, and several competitive inhibitors bound to the enzyme at the interface were determined by monitoring the ligand-conferred protection of the active site histidine residue from alkylation by phenacyl bromide. The interfacial Michaelis Menten parameters were determined from the analysis of the entire reaction progress curve and also by monitoring the effect of competitive inhibitors on the initial rate of hydrolysis in the scooting mode. The interfacial Michaelis constant (KM*) for the substrate 1,2-dimyristoylglycero-sn-3-phosphomethanol was determined to be considerably above the maximal attainable mole fraction of unity for the substrate in the bilayer. Substrate specificity studies show that the enzyme does not significantly discriminate between phospholipids that differ in the type of polar head group or in the degree of unsaturation of the fatty acyl chains. Competitive inhibitors are described that display a high degree of selectivity for binding to the nonpancreatic versus pancreatic phospholipase A2. The kinetic properties of the human nonpancreatic secreted phospholipase A2 suggest that the enzyme has evolved to hydrolyze substrates at anionic interfaces and at high calcium concentrations. PMID- 8422369 TI - Role of lateral phase separation in the modulation of phospholipase A2 activity. AB - Phospholipase A2-catalyzed hydrolysis of phosphatidylcholine large unilamellar vesicles is characterized by a period of slow hydrolysis followed by a rapid increase in the rate of hydrolysis. The temporal relationship between the burst of PLA2 activity and the lateral distribution of substrate and product lipids was examined by simultaneously recording product accumulation and the fluorescence of 1-pyrenyldecanoate, a fatty acid derivative sensitive to lipid distribution and lateral diffusion. The excimer: monomer ratio of the probe changes slowly prior to the burst in activity and then abruptly at the time of the burst. A partial phase diagram for the ternary codispersion of substrate and products (dipalmitoylphosphatidylcholine and 1:1 monopalmitoylphosphatidylcholine/palmitic acid) was constructed by differential scanning calorimetry and suggests gel/gel immiscibility in this system. Thus, the changes in pyrene fluorescence during the time course of hydrolysis appear to be due to lateral phase separation. The critical mole fraction of product both for lateral phase separation in the gel state and for elimination of the lag phase is approximately 0.083. The simultaneous recordings of PLA2 activity and pyrene fluorescence show that the lateral rearrangement of lipids begins prior to and continues during the rapid activation process of PLA2. Two possible effects of lateral phase separation are that concentration of the protein in the product-rich regions promotes putative dimerization or that formation of phase interface regions promotes enzyme activation. PMID- 8422370 TI - Lung surfactant proteins, SP-B and SP-C, alter the thermodynamic properties of phospholipid membranes: a differential calorimetry study. AB - The ability of the low molecular weight lung surfactant-associated proteins, SP-B and SP-C, to alter the thermotropic properties of synthetic multilamellar vesicles was tested using differential scanning calorimetry (DSC). The presence of either SP-B or SP-C in dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosphatidylglycerol (DPPG) multilamellar vesicles broadened the DSC thermogram and reduced the enthalpy of transition in a concentration-dependent manner. With both proteins, the temperature at which the peak of the phase transition (Tm) was detected was shifted to a higher value. The increase in Tm caused by both proteins was greater with DPPG than DPPC. We have interpreted these results as implying the presence of a protein-perturbed domain of lipid. Both SP-B and SP-C were found to influence the surface activity of the phospholipids in a concentration-dependent fashion. We speculate that instability of lipid packing predicted to occur at protein-created lipid domain boundaries may be important for the expression of surface activity in pulmonary surfactant. PMID- 8422371 TI - Thermodynamics of G-tetraplex formation by telomeric DNAs. AB - Telomeres are structures at the ends of eukaryotic chromosomes, the DNA of which contains stretches of tandemly repeated sequences with G clusters along one strand. Model telomeric G-rich DNAs can form different tetraplex structures, stabilized by cyclic hydrogen bonding of four guanines in the presence of metal ions such as Na+ or K+. Oligonucleotides with a single copy of the Oxytricha sequence dT4G4 form a tetramer, with a parallel-stranded, right-handed helical structure. Additional copies favor folded-back structures that associate to form an antiparallel dimer. The parallel-stranded tetramer has all G's in the anti configuration, while the folded-back dimer has alternating syn and anti nucleotide conformations along each strand. Here we have constructed two G tetraplex structures, containing identical G-tetrad base pairs, from oligonucleotides. One has the truncated telomeric sequence from Oxytricha, dG4T4G4, which forms an antiparallel G-quartet structure; the second is constrained to form a parallel G-strand arrangement by insertion of a 5'-p-5' linkage between two dT2G4 sequences. Each oligomer forms a defined G-tetraplex dimeric structure in the presence of Na+. The standard-state enthalpies, entropies, and free energy for formation of these tetraplexes have been determined. The parallel strand structure is thermodynamically more stable than the antiparallel one, primarily because of both greater enthalpy and entropy of formation. In addition, the two molecules differ in their interaction with sodium ions, reflecting a difference in ion binding and therefore in structure between the two forms. PMID- 8422372 TI - 13C NMR study of the effects of mutation on the tryptophan dynamics in chymotrypsin inhibitor 2: correlations with structure and stability. AB - Recombinant chymotrypsin inhibitor 2 (CI-2) and the three mutants Ile39-->Val, Ile39-->Leu, and Arg67-->Ala were successfully enriched with [2-13C]tryptophan at position 24 within the hydrophobic core of the protein. Carbon-13 NMR relaxation measurements were then used to investigate the effect of these mutations on the dynamics of the tryptophan residue. In addition, the stability of wild-type and mutant CI-2s was measured by their susceptibility to unfolding by guanidine hydrochloride. The mutant proteins were all found to be less stable, giving delta delta GU values relative to wild-type of 1.17, 1.96, and 1.21 kcal mol-1, respectively. The indole moiety of the tryptophan residue was found to be more mobile in all the mutants studied than in wild-type CI-2. Order parameters of 0.69, 0.60, 0.56, and 0.44 were derived for wild-type, Ile39-->Val, Ile39-->Leu, and Arg67-->Ala CI-2, respectively. It is concluded that there is a correlation between the protein stability and the picosecond dynamics within the hydrophobic core and that mutations can influence the dynamic behavior of the residues that are relatively distant in the three-dimensional structure. PMID- 8422373 TI - Near-UV circular dichroism of band 3. Evidence for intradomain conformational changes and interdomain interactions. AB - Near-UV circular dichroism (CD) was used to identify differences in the tertiary structure of human erythrocyte band 3, the chloride/bicarbonate exchange protein, consequent to covalent binding of anion transport inhibitors to the intramonomeric stilbenedisulfonate (ISD) site. Isolated intact band 3 and its membrane domain (B3MD) were compared. Spectral differences were observed which involved intradomain effects, in that they were seen both with intact band 3 and with B3MD, or interdomain effects, in that they were observed only for B3MD, but were inhibited when the cytoplasmic domain was attached. The intradomain effect involved a significant loss in optical activity in the Phe/Tyr region of the spectrum below 280 nm. It was seen only when the ISD site had stilbenedisulfonates bound covalently at pH 7.4. Raising the pH to 9.6 after adduct formation "normalized" this spectral change irreversibly. The interdomain effect was identified in the Trp spectral region at 292 nm. There was a significant increase in optical activity at 292 nm when bulky covalent ligands such as DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonate) were bound to B3MD, but not when the same ligands were bound to intact band 3. These latter results offer evidence that certain aspects of the conformational response of the integral domain are inhibited by the presence of an attached cytoplasmic domain. The potential significance of interdomain interactions to band 3 function is discussed briefly. PMID- 8422374 TI - A partially folded state of hen egg white lysozyme in trifluoroethanol: structural characterization and implications for protein folding. AB - The effect of 2,2,2-trifluoroethanol (TFE) on the solution conformation of hen egg white lysozyme has been investigated using circular dichroism (CD) and 1H nuclear magnetic resonance (NMR) spectroscopy. Addition of TFE to lysozyme at pH 2.0, 27 degrees C, up to a concentration of 15% (v/v) induces only slight changes in the NMR spectrum. However, above this concentration a cooperative transition to a new but partially structured state of the protein is observed. This state shows no structural cooperativity against further denaturation and is characterized by an ellipticity in the far-UV CD greater than that of the native protein. Near-UV CD intensity is dramatically reduced compared with that of the native state, and 1H NMR studies indicate that side-chain interactions are substantially averaged in this denatured state. Solvent proton/deuterium exchange rates for 66 amide hydrogens were measured site-specifically by a combination of amide trapping experiments and 2D 1H NMR. Significant protection from exchange occurs for about 25 backbone amides, the majority of which are located in regions of the protein that are helical in the native enzyme. By contrast, amides located in a second region of the native protein which contains a beta-sheet and one 3(10)-helix as well as a long loop show little protection. This pattern of protection resembles that found in the stable molten globule state of alpha lactalbumin and in an early kinetic intermediate detected in the refolding of hen lysozyme. PMID- 8422375 TI - Use of hydrazine to release in intact and unreduced form both N- and O-linked oligosaccharides from glycoproteins. AB - The use of hydrazine to release unreduced N- and O-linked oligosaccharides from glycoproteins has been investigated using several "standard" glycoproteins of previously defined glycosylation. It is shown that hydrazinolysis can be used to release intact N- and O-linked oligosaccharides in an unreduced form. The release of O-linked oligosaccharides occurs with a lower temperature dependence than the release of N-linked oligosaccharides, and the kinetic parameters governing release of oligosaccharides from these standard glycoproteins have been determined. These parameters allow a definition of reaction conditions under which anhydrous hydrazinolysis can be used to selectively release O-linked oligosaccharides (60 degrees C, 5 h) or release both N- and O-linked oligosaccharides (95 degrees C, 4 h) in high yield (> 85%) from all glycoproteins investigated (n = 11). Under these reaction conditions, the recovered N- and O linked oligosaccharides are structurally intact (as judged by 600-MHz 1H-NMR, laser-desorption mass spectrometry, HPAEC-PAD, gel filtration, and glycosidase digestion), with the possible exception of certain N- and O-acyl substituents of sialic acid. This use of mild hydrazinolysis therefore allows both the simultaneous and sequential chemical release from glycoproteins of O- and N linked oligosaccharides in their intact unreduced form. PMID- 8422376 TI - Binding of substrates to human deoxycytidine kinase studied with ligand-dependent quenching of enzyme intrinsic fluorescence. AB - Deoxycytidine kinase is a key enzyme in the salvage pathway, and its activity is required for 5'-phosphorylation of several important antiviral and cytostatic nucleoside analogues. It has recently been purified completely from human sources. Steady-state and time-resolved fluorescence of human deoxycytidine kinase was used to study its interaction with the substrates dCyd, dAdo, dUrd, dTTP, and the feedback inhibitor dCTP. Enzyme fluorescence quenching by dCTP, dCyd, dTTP, and dAdo was bimodal, and the best fits of the quenching patterns were obtained using two modified Stern-Volmer equations with two sets of quenching constants (Ksv) and accessibility values (fa) fitted independently for "low" and "high" concentration ranges of ligands. The transition between these occurred at about 20 microM dCTP, 50 microM dCyd, 30 microM dTTP, and 180 microM dAdo. Enzyme fluorescence showed unimodal quenching by dAdo and 30% reduced accessibility of the binding site in the presence of dCyd. dUrd quenching was also unimodal with Ksv = 0.0047 +/- 0.0007 microM-1 and fa = 0.75 +/- 0.05, hence in the same range as for the "high" concentration range of dAdo in the absence of dCyd, where they are 0.0025 +/- 0.0003 microM-1 and 0.73 +/- 0.03, respectively. Fluorescence quenching was used to directly determine enzyme-ligand binding and revealed bimodal binding of dCTP, dCyd, dTTP, and dAdo and unimodal binding of dUrd, and of dAdo in the presence of 0.1 microM dCyd. Transition between these two modes of binding occurred at the concentrations described above.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422377 TI - Characterization of an intermediate in the folding pathway of phosphoglycerate kinase: chemical reactivity of genetically introduced cysteinyl residues during the folding process. AB - The unfolding-refolding kinetics of yeast phosphoglycerate kinase were studied using the chemical reactivity of genetically introduced cysteinyl residues as conformational probes and far-ultraviolet circular dichroism. A unique internal cysteinyl residue was introduced in several mutants at selected positions in the N- and C-domains. The cysteinyl residues were at positions 97 (the unique cysteinyl residue of the wild-type enzyme), 183 in the N-domain, 285 and 324 in the C-domain. A similar strategy has been used to study the unfolding-refolding transition under equilibrium conditions [Ballery et al. (1990) Protein Eng. 3, 199-204]. Except for the mutant C97A,A183C, whose cysteinyl residue is located at the domain interface, three labeling phases were observed during the refolding process, indicating the presence of three species, the unfolded, intermediate, and folded proteins. The comparison of the data obtained following the accessibility of the thiol group to 5,5'-dithiobis(2-nitrobenzoate) and ellipticity at 218 nm indicated that all mutants have the same folding pathway and allowed us to characterize the intermediate. In this species, each domain appeared to have a high content of secondary structure but a flexible tertiary structure; this intermediate, which had the characteristics of a molten globule, remained in fluctuating equilibrium with a widely unfolded form. The same folding intermediate was detected for mutant C97A,A183C; however, the cysteinyl residue being totally accessible to the reagent, it is likely that in this intermediate the interdomain interactions are not established. Domain pairing and formation of the native tertiary structure occur simultaneously in the slow phase of refolding. The validity and limitations of the methodology are discussed. PMID- 8422378 TI - Sequence-specific deamidation: isolation and biochemical characterization of succinimide intermediates of recombinant hirudin. AB - Natural hirudin variant 2 with a lysine residue in position 47 (rHV2-Lys47) was produced in a genetically engineered strain of Saccharomyces cerevisiae as a secreted protein of 65 amino acids and purified to greater than 99% homogeneity. Only reversed-phase high-performance liquid chromatography (RP-HPLC) using very shallow acetonitrile gradients indicated the presence of a component in the final product (approximately 1% of total protein) with a slightly increased retention time. Using successive RP-HPLC purification steps, this hydrophobic impurity was isolated and separated into two constituents defined as components A1 and A2 which differed from the parent molecule by mass reductions of 17.2 Da (A1) and 17.6 Da (A2), respectively, as determined by electrospray mass spectrometry (ESMS). Proteolytic digestion with endoprotease Glu-C from Staphylococcus aureus (V8 protease) and analysis of the peptide mixture by ESMS showed that the mass difference between rHV2-Lys47 and component A1 was due to a modification between amino acids 1 and 43, while the corresponding mass difference with component A2 was the result of a modification within the peptide fragment comprising residues 50-61. Further analyses using amino acid sequencing and ESMS in combination with collision-activated dissociation (CAD) detected modifications at residues Asn33 Gly34 in component A1 and at Asn53-Gly54 in component A2. Both of these sites were previously shown to be susceptible to spontaneous deamidation under slightly basic pH conditions. Thus, the mass reductions of approximately 17 Da and the fact that both asparagines, Asn33 in component A1 and Asn53 in component A2, proved to be resistant to Edman degradation provided strong support for them being stable succinimide intermediates of the corresponding deamidation reactions. Both intermediates were shown to have inhibition constants for human alpha-thrombin on the order of 1 pM, identical to that of rHV2-Lys47. The isoelectric point of component A2 was determined to be within 0.01 pH unit of that of the parent molecule by isoelectric focusing in an immobilized pH gradient. PMID- 8422379 TI - Horse liver alcohol dehydrogenase-catalyzed oxidation of aldehydes: dismutation precedes net production of reduced nicotinamide adenine dinucleotide. AB - The oxidation of aldehydes by horse liver alcohol dehydrogenase (HL-ADH) is more complex than previously recognized. At low enzyme concentrations and/or high aldehyde concentrations, a pronounced lag in the assay progress curve is observed when the reaction is monitored for NADH production at 340 nm. When the progress of the reaction is followed by 1H NMR spectroscopy, rapid dismutation of the aldehyde substrate into the corresponding acid and alcohol is observed during the lag phase. Steady-state production of NADH commences only after aldehyde concentrations drop below 5% of their initial value; thereafter, NADH production occurs with continuous adjustment of the equilibrium between aldehyde, alcohol, NADH, and NAD+. The steady-state NADH production exhibits normal Michaelis-Menten kinetics and is in accord with earlier studies using much higher enzyme concentrations where no lag phase was reported. These results establish that the ability of HL-ADH to oxidize aldehydes is much greater than previously thought. The relationship between aldehyde dismutase and aldehyde dehydrogenase activities of HL-ADH is discussed. PMID- 8422380 TI - Structures of the modified folates in the thermophilic archaebacteria Pyrococcus furiosus. AB - The structures of the modified folates present in Pyrococcus furiosus have been determined. This was accomplished largely by the characterization of the arylamines resulting from the air oxidative cleavage of the reduced modified folates present in these cells, using both chemical and enzymatic methods. Cell extracts separated on DEAE-Sephadex columns showed one major peak containing the arylamines derived from the modified folates. These arylamines were not retained on the DEAE-Sephadex columns, indicating that they contained no net negative charge. Purification of the azo dye derivatives of these arylamines on a Bio-Gel P-6 column showed the presence of three different compounds (compounds 1, 2, and 3) in an average amount of 4.1, 7.6, and 22 nmol/g dry weight of cells, respectively. Each of these compounds readily underwent mild acid hydrolysis (0.1 M HCl, 110 degrees C, 1 min) to produce the azo dye derivative of 5-(p aminophenyl)-1,2,3,4-tetrahydroxypentane (pAPT). The structure and stereochemistry (ribo) of the pAPT was the same as the pAPT present in methanopterin. In addition, compounds 1, 2, and 3 were each shown to contain 1 mol equiv of ribose and 1, 2, and 3 mol equiv of N-acetylglucosamine (gluNAc), respectively, and were designated as the azo dye derivatives of pAPT-ribose gluNAc, pAPT-ribose-(gluNAc)2, and pAPT-ribose-(gluNAc)3. Each of these compounds was readily cleaved to the azo dye derivative of pAPT-ribose by the enzymatic action of beta-N-acetylglucosaminidase, indicating that all the gluNAc residues were beta-linked.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422381 TI - Structure of the asn-linked oligosaccharides of apolipophorin III from the insect Locusta migratoria. Carbohydrate-linked 2-aminoethylphosphonate as a constituent of a glycoprotein. AB - The primary structures of the N-linked carbohydrate chains of apolipophorin III from the insect Locusta migratoria have been determined. The glycoprotein was completely deglycosylated with peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F. Released oligosaccharides were separated from the remaining protein using gel-permeation chromatography on Bio-Gel P-100. Purification of the carbohydrate chains was achieved by a combination of FPLC anion-exchange chromatography on Mono-Q and amine adsorption HPLC on Lichrosorb-NH2. The structures of the carbohydrate chains were deduced with a combination of fast atom bombardment mass spectrometry, 1H- and 31P-NMR spectroscopy, and methylation analysis. The majority of the carbohydrate chains contains 2 aminoethylphosphonate (AEP), which is linked to the 6-position of Man and/or GlcNAc. L. migratoria apolipophorin III is the first example of a glycoprotein containing carbohydrate-linked 2-aminoethylphosphonate. The structures of the major oligosaccharides were established to be the following: [formula: see text] PMID- 8422382 TI - Inhibition of bovine lens leucine aminopeptidase by bestatin: number of binding sites and slow binding of this inhibitor. AB - Aminopeptidases catalyze the hydrolysis of amino acid residues from the amino terminus of peptide substrates. Their activity has been implicated in myriad fundamental biochemical and physiological processes, and alterations in aminopeptidase activity have been correlated with a variety of pathologies. Nevertheless, information about this group of proteases is less well developed. Bovine lens leucine aminopeptidase (blLAP) can be considered prototypical of many enzymes in this family of peptidases. It shows common features of (1) requiring divalent metal ions for activity, (2) having a relatively large size, and (3) having slow, relatively tight binding of bestatin, a transition-state analog of the substrate PheLeu. Bovine lens LAP is the only bestatin-inhibitable aminopeptidase for which structural and mechanistic data are available. However, full exploitation of these data required knowledge of the number of inhibitor molecules bound per subunit. Independent direct binding experiments and kinetic determinations indicate that one bestatin is bound per subunit in blLAP. Ki and Ki* for formation of the initial and final complexes are approximately 1.1 x 10( 7) and 1.3 x 10(-9) M, respectively. The mode of binding is slow and competitive. The t1/2 for formation and deformation of the final enzyme-inhibitor complex is approximately 30 and 22 min, respectively, with 10(-8) M bestatin. To perform these measures, a new assay using physiological peptides (LeuGlyGly) as substrate was adapted. Taken together with prior NMR, photoaffinity labeling, and crystallographic data, these binding data allow us to propose a mechanism of the blLAP-catalyzed hydrolysis of peptides. PMID- 8422383 TI - Halophilic class I aldolase and glyceraldehyde-3-phosphate dehydrogenase: some salt-dependent structural features. AB - Aldolase and glyceraldehyde-3-phosphate dehydrogenase from the extremely halophilic archaebacterium Haloarcula vallismortis are stable only in high concentrations of KCl present within the physiological environment. Data concerning the structural changes in the two enzymes as a result of lowering of salt concentration and changes in pH were obtained by monitoring the intrinsic protein fluorescence in the presence of quenchers. When the KCl concentrations were lowered below 2 M or in the presence of 6 M guanidine hydrochloride, the emission maximum shifted to a longer wavelength, indicating enhanced exposure of tryptophyl residues to the solvent. The spectral characteristics of the two proteins in guanidine hydrochloride and 0.4 M KCl were identical. However, these denatured states appear to be different than those observed after acid denaturation. Further perturbation of fluorescence was observed due to I-, and application of the Stern-Volmer law showed that the total fluorescence was available to the quenchers only in 0.4 M KCl solutions. The unfolding of proteins in 0.4 M KCl was a gradual process which was accompanied by a time-dependent loss in enzyme activity. The activity loss was complete within 30 min for aldolase whereas in the case of GAPDH nearly 3 h was required for the destruction of activity. For both enzymes, inactivation and protein denaturation were strongly correlated. The data on activity and thermostability measurements of the two enzymes in varying concentrations of KCl and potassium phosphate revealed that though both proteins are halophilic, the forces in the maintenance of their stability could be different.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422384 TI - Escherichia coli fumarase A catalyzes the isomerization of enol and keto oxalacetic acid. AB - Fumarase A, a product of the fumA gene of Escherichia coli, has been found to catalyze the isomerization of enol to keto oxalacetic acid (OAA) in addition to catalyzing the fumarase reaction. The kcat/Km for the isomerization is almost identical to that for the fumarase reaction. The isomerization reaction apparently takes place at the same active site as the fumarase reaction since both reactions show a similar sensitivity to inactivation by O2, both reactions are strongly inhibited by 2-hydroxy-3-nitropropionate, and the isomerization reaction is inhibited by fumarate and malate. The isomerization requires the presence of a [4Fe-4S] or [3Fe-4S] cluster, perhaps for structural rather than catalytic reasons. Hydration of enol OAA to the gem diol has been ruled out as a possible mechanism of isomerization on the basis of the preservation of the oxygen on carbon 2 and the position of protonation on carbon 3. The isomerization is not stereospecific in the position of protonation at carbon 3 but appears to be stereoselective, with protonation preferentially occurring in the 3-pro-S position. Porcine fumarase, isopropyl malate isomerase, and dihydroxyacid dehydratase do not catalyze this isomerization. Fumarase A and aconitase, two enzymes with 4Fe-4S clusters that bind a linear 4-carbon dicarboxylic acid moiety in the trans conformation during their normal hydro-lyase reaction, do catalyze this isomerization. PMID- 8422385 TI - Inhibition of tryptophan synthase by (1-fluorovinyl)glycine. AB - Tryptophan synthase (alpha 2 beta 2 complex) from Salmonella typhimurium catalyzes the formation of tryptophan from serine and indole. The enzyme is inactivated by (1-fluorovinyl)glycine. Concomitant with enzyme inactivation, the absorbance at 485 nm increases, indicating covalent modification of pyridoxal 5' phosphate. It is proposed that inactivation involves elimination of HF to form an allene, which reacts with a nucleophile at the active site. The inactivation reaction involves an alpha,beta-elimination, as does the formation of tryptophan from indole and serine. The inactivation occurs with k(in) > 1.3 s-1, which is very close to k(cat) (6.4 s-1) for the formation of tryptophan from indole and serine. The inactive enzyme (alpha 2 beta 2) regains activity with k(off) = 0.005 min-1. Aminoacetone is formed during reaction, and pyridoxal 5'-phosphate is regenerated. Tryptophan synthase also catalyzes the dehydration of serine, or 3 fluoroalanine, to pyruvate in the absence of indole. This reaction involves an alpha,beta-elimination and the intermediate formation of an aminoacrylate adduct with pyridoxal 5'-phosphate, as does the formation of tryptophan. Pyruvate formation proceeds at less than 5% the rate of tryptophan formation. With [2 2H]serine an isotope effect (DVmax = 1.5) is observed. We propose that pyruvate formation is limited by the rate of hydration of the aminoacrylate intermediate and the rate of the abstraction of the serine alpha-hydrogen. PMID- 8422386 TI - Rat liver aromatic L-amino acid decarboxylase: spectroscopic and kinetic analysis of the coenzyme and reaction intermediates. AB - The physiochemical properties of the coenzyme in rat liver aromatic L-amino acid decarboxylase (AADC) expressed in Escherichia coli have been studied by spectroscopic analysis of the enzyme, its reaction intermediates, and its complexes with substrate analogs. The enzyme, having one pyridoxal 5'-phosphate (PLP) per subunit, shows a prominent absorption maximum at 335 nm and a weaker one at 425 nm. The spectrum did not essentially change in the pH range of 6.0 8.0. When the coenzyme was excited at 335 nm, it emitted fluorescence primarily at 520 nm. The structure for the absorption at 335 nm was ascribed to the enolimine form of the PLP-lysine Schiff base. On the reaction of AADC with L-3,4 dihydroxyphenylalanine (L-dopa), the absorption of PLP showed biphasic changes before reaching a steady-state. Results of both pre-steady-state and steady-state kinetic analyses were consistent with the model that the reaction proceeds as shown in the equation: E + S<==>X1<==>X2-->E + P. The rate constant was determined for each step, and the Km value for L-dopa was obtained as 0.086 mM. The absorption spectra of the two intermediates, X1 and X2, were postulated from the calculation of the absorption changes during the first and the second steps of the reaction in which X1 and X2 showed an absorption maximum at 425 and 380 nm, respectively, with a concomitant decrease in absorbance at 335 nm. These predicted absorption spectra of X1 and X2 showed striking resemblances to those of AADC complexed with dihydroxyphenylacetic acid (DOPAc) and L-dopa methyl ester (DopaOMe), respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422387 TI - Antagonistic action of imidazolineoxyl N-oxides against endothelium-derived relaxing factor/.NO through a radical reaction. AB - A labile inorganic free radical, nitric oxide (.NO), is produced by nitric oxide synthase from the substrate L-arginine in various cells and tissues. It acts as an endothelium-derived relaxing factor (EDRF) or as a neurotransmitter in vivo. We investigated the reactivity of stable radical compounds, imidazolineoxyl N oxides such as 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO), carboxy-PTIO, and carboxymethoxy-PTIO against .NO/EDRF in both chemical and biological systems. By using electron spin resonance (ESR) spectroscopy, imidazolineoxyl N-oxides were found to react with .NO in a stoichiometric manner (PTIO/.NO = 1.0) in a neutral solution (sodium phosphate buffer, pH 7.4) with rate constants of approximately 10(4) M-1 s-1, resulting in the generation of NO2 /NO3- and imidazolineoxyls such as 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl (PTI), carboxy-PTI, or carboxymethoxy-PTI. Furthermore, the effects of imidazolineoxyl N-oxides on acetylcholine- or ATP-induced relaxation of the smooth muscle of rabbit aorta were tested. The vasorelaxations were inhibited by all three imidazolineoxyl N-oxides markedly. The inhibitory effects of carboxy PTIO was almost 2-fold stronger than those of .NO synthesis inhibitors, N omega nitro-L-arginine and N omega-monomethyl-L-arginine. Generation of EDRF/.NO was identified by reacting the PTIO in aortic strips and quantitating the reaction product with ESR spectroscopy. Thus, it was clarified that imidazolineoxyl N oxide antagonize EDRF/.NO via a unique radical-radical reaction with .NO. PMID- 8422388 TI - Kinetics of the oxidation of p-coumaric acid by prostaglandin H synthase and hydrogen peroxide. AB - Steady-state kinetics of the oxidation of p-coumaric acid (CA) by prostaglandin H synthase and hydrogen peroxide was studied at 25 degrees C in 0.1 M phosphate buffer, pH 8.0, using a stopped-flow apparatus. The following evidence supports a mechanism in which CA serves as a reducing substrate for prostaglandin H synthase through two one-electron oxidation steps: (a) the oxidation product of CA is the same in the prostaglandin H synthase/hydrogen peroxide and the horseradish peroxidase/hydrogen peroxide systems; (b) an identical steady-state enzyme intermediate (compound II) is present in both systems; (c) CA stimulates the cyclooxygenase activity of prostaglandin H synthase; the concentration of CA that produces 50% stimulation, A50, is 350 +/- 30 microM. On the time scale of our experiments, the inactivation of prostaglandin H synthase by hydrogen peroxide was insignificant when CA was present. A molar absorptivity of 17.2 +/- 0.9 mM-1 cm-1 at 300 nm was determined for CA which was used to follow the initial rate of disappearance of CA. The reaction of CA with hydrogen peroxide catalyzed by prostaglandin H synthase showed saturation behavior. An irreversible reaction mechanism for the steady-state kinetics of prostaglandin H synthase is proposed which is consistent with all of our experimental results. Under steady-state conditions, the second-order rate constants for the reactions of prostaglandin H synthase with hydrogen peroxide and prostaglandin H synthase-compound II with CA are (9.2 +/- 0.1) x 10(5) and (2.5 +/- 0.1) x 10(6) M-1 s-1, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422389 TI - Charge recombination between P700+ and A1- occurs directly to the ground state of P700 in a photosystem I core devoid of FX, FB, and FA. AB - The charge recombination between P700+ and electron acceptor A1- was studied by flash kinetic spectroscopy in a photosystem I core devoid of iron-sulfur centers FX, FB, and FA. We showed previously that the majority of the flash-induced absorption change at 820 nm decayed with a 10-microseconds half-time, which we assigned to the disappearance of the P700 triplet formed from the backreaction of P700+ with A1- [Warren, P.V., Parrett, K.G., Warden, J.T., & Golbeck, J.H. (1990) Biochemistry 29, 6545-6550]. We have reinvestigated this assignment in the near UV, blue, and near-IR wavelength regions. The difference spectrum from 380 to 480 nm and from 720 to 910 nm shows that the P700+ A1- charge recombination is dominated by the P700 cation rather than the P700 triplet. Accordingly, the 10 microseconds kinetic transient represents the direct backreaction of P700+ with A1-, which repopulates the ground state of P700. This is unlike a P700-FA/FB complex where, in the presence of reduced FX-, FB-, and FA-, the P700+ A1- charge recombination populates the P700 triplet state [Setif, P., & Bottin, H. (1989) Biochemistry 28, 2689-2697]. The A1 acceptor is highly susceptible to disruption by detergents in the absence of iron-sulfur center FX. The addition of 0.1% Triton X-100 to the P700-A1 core leads to a approximately 2.5-fold increase in the magnitude of the flash-induced absorption change at 780 nm; thereafter, 85% of the absorption change decays with a 25-ns half-time and 15% decays with a 3 microseconds half-time.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422391 TI - Time-resolved infrared spectral analysis of the KL-to-L conversion in the photocycle of bacteriorhodopsin. AB - Time-resolved infrared spectra of the hydrated film of light-adapted bacteriorhodopsin were recorded from earlier than 200 ns to 450 microseconds after light excitation in the 1800-900-cm-1 region on a newly designed dispersive type infrared spectrometer [Iwata & Hamaguchi (1989) Appl. Spectrosc. 44, 1431 1437]. Both the KL-to-L and L-to-M conversions were detected in this time range. The spectral shape of KL is similar to K measured at 77 K except for the intense hydrogen out-of-plane vibrational band at 984 cm-1. The kinetics of this band are different from those of the other KL-specific bands at 1510, 1296, and 956 cm-1. Since the hydrogen out-of-plane vibrational bands are intensified by twists of the polyene chain, a change in the twist of the chromophore is suggested within the lifetime of KL. During the decaying process of L, the KL-specific vibrational bands are observed in parallel with L, indicating that KL and L are in equilibrium. PMID- 8422390 TI - Effects of phospholipase A2 digestion on the carotenoid and bacteriochlorophyll components of the light-harvesting complexes in Rhodobacter sphaeroides chromatophores. AB - The instantaneous electrochromic response of carotenoids associated with the B800 850 light-harvesting complex of Rhodobacter sphaeroides has been used widely as an intrinsic probe of membrane potential. In the present study, the structural basis for this phenomenon was examined by phospholipase A2 digestion of chromatophores from R. sphaeroides strain NF57G, containing B800-850 as the sole pigment-protein complex. The major phospholipase-induced alterations of the overall carotenoid absorption spectrum were characterized by an absorbance loss and a blue shift that were accompanied by a decrease in absorbance at 800 nm and a red shift in the B850 absorbance band. In wild-type chromatophores, the electrochromic carotenoid response induced by both flash illumination and a K+ diffusion potential was diminished by approximately 60% after 1 h of digestion. The initial loss of the carotenoid response was correlated specifically to the hydrolysis of phosphatidylethanolamine, and was shown to arise from effects exerted directly upon the electrochromically active carotenoid pool, possibly by alterations in the spatial relationship between the field-sensitive carotenoids and the polarizing permanent field. In phospholipase A2-digested NF57G preparations in which the B800 band was diminished by nearly half and the carotenoid response was abolished, no significant changes in the efficiency of energy transfer from carotenoids to bacteriochlorophyll were detected at 77 K, suggesting that the electrochromically active carotenoids are not energetically linked to B800 bacteriochlorophyll. PMID- 8422392 TI - Ligand-protein electrostatic interactions govern the specificity of retinol- and fatty acid-binding proteins. AB - Cellular retinol-binding protein II (CRBP-II) and intestinal fatty acid-binding protein (I-FABP) are both expressed in small intestinal enterocytes and exhibit 31% sequence identity. I-FABP binds a single molecule of long-chain fatty acid and forms an ion-pair electrostatic interaction between the cationic side chain of arginine-106 and the anionic fatty acid carboxyl group. In contrast, CRBP-II binds all-trans-retinol or -retinal and contains a glutamine residue in the corresponding position, residue 109. We have characterized and compared the interactions of fatty acids and retinoids with I-FABP, CRBP-II, and two reciprocal mutant proteins. The mutants were designated CRBP-II(Q109R), where glutamine-109 was replaced by arginine, and I-FABP(R106Q), where arginine-106 was replaced by glutamine. As monitored by titration calorimetry and carbon-13 NMR spectroscopy, the fatty acid-binding properties of CRBP-II(Q109R) were found to be essentially identical to those of wild-type I-FABP. Both proteins bound 1 molecule of fatty acid with identical affinities (Kd = 0.2 microM). The enthalpic contribution to the total free energy of binding was large for both proteins: 66% and 87%, respectively. In addition, the carboxyl groups of fatty acids bound to both proteins were solvent-inaccessible. There was little or no change in the ionization state of the bound fatty acid over a wide pH range, as monitored by the chemical shift of the fatty acid carboxyl 13C resonance. Furthermore, the binding of fatty acid to both proteins was accompanied by a selective perturbation of the guanidino 13C resonance of a single arginine residue.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422393 TI - Quantitative analysis of liposome-cell interactions in vitro: rate constants of binding and endocytosis with suspension and adherent J774 cells and human monocytes. AB - We have characterized the parameters describing the total association (uptake) of liposomes with murine macrophage-like cell line J774 cells and human peripheral blood monocytes at 4 degrees C and at 37 degrees C with or without inhibitors of endocytosis. The uptake of neutral liposomes composed of phosphatidylcholine (PC)/cholesterol (Chol) (2:1 mole ratio) is about 10-fold lower than that of negatively charged liposomes composed of phosphatidylserine (PS)/PC/Chol (1:1:1). However, the rate of uptake of PC/Chol liposomes at 37 degrees C is still 10-fold higher than that by fluid-phase pinocytosis. The uptake of liposomes, which is mediated by high-affinity binding to the cell surface binding sites and subsequent endocytosis, could be simulated and predicted by model calculations employing mass action kinetics. The number of binding sites, affinity constants of binding at 37 degrees C and 4 degrees C, on- and off-rate constants of binding, and endocytic rate constants for both types of liposomes were determined. The number of binding sites and the binding constants for PS/PC/Chol liposomes binding to J774 cells is severalfold to an order of magnitude higher than that for PC/Chol liposomes, but the rate constants at which they are endocytosed following binding to the cells are similar for both liposome types. The binding of liposomes, especially PS/PC/Chol, to J774 cells and monocytes is greatly enhanced by adherence of cells to plastic substratum and is also increased by maturation/differentiation in the case of monocytes. Our quantitative analysis indicates that the binding and endocytosis of liposomes, especially PS-containing liposomes, is mediated by binding sites that have strong affinity, comparable to or about an order of magnitude smaller than other known particle-cell interactions with specific receptors such as virus and lipoproteins binding to cells. PMID- 8422394 TI - Inactivation of PR8 influenza virus through the octadecylrhodamine B chloride membrane marker. AB - The octadecylrhodamine B chloride (R18) membrane marker was incorporated into PR8 influenza viruses and virus receptor (GD1a-) containing small unilamellar vesicles (SUV). Both were tested in a fusion/lipid transfer assay [Wunderli Allenspach, H., & Ott, S. (1990) Biochemistry 29, 1990-1997] to find out whether incorporation into artificial and biological membranes yields equivalent results. The R18 assay is based on incorporation of quenched concentrations of the label into donor membranes and monitoring of the dequenching upon its dilution into unlabeled acceptors. With PR8 viruses and R18-labeled SUV, a fast, hemagglutinin specific fusion takes place at pH 5.3, independently of the initial quenching. At neutral pH, a slow, nonspecific R18 transfer occurs. Both processes follow second order kinetics. Upon incubation of R18-labeled PR8 viruses with unlabeled SUV or LUV in neutral buffer, transfer is also found. At pH 5.3, a complex dequenching curve best described with superposition of two second-order functions was encountered: a fast, hemagglutinin-specific component and a slow, nonspecific component. A decreasing proportion of the fast fusion was found with increasing initial quenching of labeled virus. Extrapolation showed that full fusion activity is obtained only with low initial quenching (20-30%). The gradual inactivation of the virus by increasing amounts of R18 was confirmed with biological assays (e.g., infectivity). The R18 surface density is much higher in viruses than in liposomes to obtain the same initial quenching. Analysis with the Stern-Volmer plot revealed that R18 monomers and dimers contribute to the quenching in labeled PR8 viruses, whereas only dimers determine the quench curve in liposomes. PMID- 8422395 TI - Conformational analysis of [D-Ala9]alpha-factor and [L-Ala9]alpha-factor in solution and in the presence of lipid. AB - The conformations in solution and in the presence of lipid vesicles of [D-Ala9] and [L-Ala9] analogues of the alpha-factor (WHWLQLKPGQPMY) from the yeast Saccharomyces cerevisiae were examined by NMR spectroscopy. Although both peptides are flexible molecules, NOE and NH d delta/dT data indicate that the [D Ala9]alpha-factor analogue in DMSO and aqueous solution adopts a type II beta turn about residues 8 and 9. In contrast, various NMR parameters for the less active [L-Ala9] analogue do not provide evidence for a regular secondary structure in solution. Transfer NOE data indicate that for both peptides binding to the lipid is strongest for the N-terminal residues. The C-terminus of the [D Ala9] analogue appears to be more constrained in the bound state than the C terminus of the [L-Ala9] analogue. This result is consistent with transfer NOE evidence that the type II beta-turn conformation of the [D-Ala9]alpha-factor is maintained in the lipid bound state. PMID- 8422396 TI - pH dependence of the active site of horseradish peroxidase compound II. AB - Using X-ray absorption spectroscopy, we investigated the active site of horseradish peroxidase (HRP) compound II at two different pH values. The results indicate that the bond length of the sixth coordinated ligand of the active site was 1.90 +/- 0.02 A at pH 7, decreasing to 1.72 +/- 0.02 A at pH 10. The average iron-to-pyrrole nitrogen and the proximal ligand bond lengths showed no significant changes. The position of higher coordination shells around the iron center changed, implying that some movement or deformation of nearby amino acid residues and/or of the heme occurred. Results of this study suggest that the decrease of the Fe-O bond length of HRP compound II at the higher pH might be attributed to the loss of a hydrogen bond which is present between the oxygen ligand and an amino acid residue in the heme pocket at pH 7. PMID- 8422397 TI - A symmetric inhibitor binds HIV-1 protease asymmetrically. AB - Potential advantages of C2-symmetric inhibitors designed for the symmetric HIV-1 protease include high selectivity, potency, stability, and bioavailability. Pseudo-C2-symmetric monools and C2-symmetric diols, containing central hydroxymethylene and (R,R)-dihydroxyethylene moieties flanked by a variety of hydrophobic P1/P1' side chains, were studied as HIV-1 protease inhibitors. The monools and diols were synthesized in 8-10 steps from D-(+)-arabitol and D-(+) mannitol, respectively. Monools with ethyl or isobutyl P1/P1' side chains were weak inhibitors of recombinant HIV-1 protease (Ki > 10 microM), while benzyl P1/P1' side chains afforded a moderately potent inhibitor (apparent Ki = 230 nM). Diols were 100-10,000x more potent than analogous monools, and a wider range of P1/P1' side chains led to potent inhibition. Both classes of compounds exhibited lower apparent Ki values under high-salt conditions. Surprisingly, monool and diol HIV-1 protease inhibitors were potent inhibitors of porcine pepsin, a prototypical asymmetric monomeric aspartic protease. These results were evaluated in the context of the pseudosymmetric structure of monomeric aspartic proteases and their evolutionary kinship with the retroviral proteases. The X-ray crystal structure of HIV-1 protease complexed with a symmetric diol was determined at 2.6 A. Contrary to expectations, the diol binds the protease asymmetrically and exhibits 2-fold disorder in the electron density map. Molecular dynamics simulations were conducted beginning with asymmetric and symmetric HIV-1 protease/inhibitor model complexes. A more stable trajectory resulted from the asymmetric complex, in agreement with the observed asymmetric binding mode. A simple four-point model was used to argue more generally that van der Waals and electrostatic force fields can commonly lead to an asymmetric association between symmetric molecules. PMID- 8422398 TI - Sampling of the conformations of the d(CGCTGCGGC) hairpin in solution by two dimensional nuclear magnetic resonance and theoretical methods. AB - Most NMR studies of DNA oligomers have focused on rigid structures that show a strong preference for one or a small set of ground-state conformations. There is an increasing interest in extending NMR methods to investigate DNA systems in which this preference does not exist. A DNA hairpin is one such system where a large number of low-energy structures coexist in solution. In this article we show how 1D/2D NMR data of the d(C1-G2-C3-T4-G5-C6-G7-G8-C9) hairpin are used to map the conformational space of this molecule. First, we characterize the gross morphology of the hairpin by monitoring the exchangeable imino signals in the molecule. Second, we extract a set of inter-proton distances (i.e., the average values and the associated dispersions) for various pairwise interactions by performing full-matrix NOESY simulation with respect to the observed NOESY data for mixing times of 250 and 100 ms. Third, we use these distances as structural constraints to perform a 300-ps molecular dynamics simulation at 500 K. Fourth, we extract 600 snapshots (one after every 0.5 ps) from the MD trajectory and perform constrained energy minimization to map local minima on the sampled energy surface (we call this the rapid temperature quenching step). Fifth, we assign 600 structures to 14 disjoint clusters such that conformationally similar hairpins belong to the same cluster while conformationally distinct hairpins belong to different clusters. Finally, we interpret the NOESY data in terms of conformationally distinct structures by recalculating NOESY contributions taken from representative structures of different clusters. Our analyses clearly demonstrate that the NMR data correspond to an ensemble of distinct structures, i.e., a set of energetically stable but conformationally distinct structures that satisfies the constraints of loop folding in the d(C1-G2-C3-T4-G5-C6-G7-G8-C9) hairpin. Two types of loop folding consistent with NMR data are obtained: (i) a hairpin with two G.C pairs in the stem and four residues in the loop and (ii) a hairpin with two G.C pairs and a reverse wobble G.T pair in the stem plus two residues in the loop. PMID- 8422399 TI - Localized interaction of the polyamine methylspermidine with double-helical DNA as monitored by 1H NMR self-diffusion measurements. AB - The 1H NMR pulsed field gradient self-diffusion method has been used to measure the diffusion coefficient of the polyamine analogue methylspermidine (completely N-methylated spermidine) in DNA solution, as a function of the concentration ratio of methylspermidine to DNA phosphate. Three different DNA's have been investigated: d(GC)4 (8 base pairs), core length calf thymus DNA (approximately 120 base pairs), and sonicated high molecular weight calf thymus DNA (average 7500 base pairs). For a constant ratio of methylspermidine to DNA phosphate, the diffusion coefficient decreases with increasing DNA length. Moreover, at low concentration ratios the diffusion coefficient of methylspermidine approaches a limiting value that is close to that of the DNA molecule. The experimental data are well reproduced by a two-state diffusion model. In this model the diffusion coefficient of the polyamine is a population-weighted average of polyamine associated with DNA (with a diffusion coefficient given by that of the DNA molecule) and polyamine free in solution. PMID- 8422401 TI - Mutagenicity and genotoxicity of the major DNA adduct of the antitumor drug cis diamminedichloroplatinum(II). AB - The mutagenicity and genotoxicity of cis-[Pt(NH3)2[d(GpG)-N7(1),-N7(2)]] (G*G*), the major DNA adduct of the antitumor drug cisplatin, has been investigated in Escherichia coli. A duplex bacteriophage M13 genome was constructed to contain the G*G* adduct at a specific site in the (-) strand. The singly platinated duplex genome exhibited a survival of 22% relative to that of the unplatinated control genomes, and this value rose to 38% in cells treated with ultraviolet light to induce the SOS response. Singly platinated single-stranded genomes were also produced. Replication of the single- and double-stranded genomes in vivo yielded SOS-dependent, targeted mutations at frequencies of 1.3% and 0.16%, respectively. The mutagenic specificity of G*G* in both single- and double stranded DNA was striking in that 80-90% of the mutations occurred at the 5' platinated G. Approximately 80% of the mutations were G-->T transversions at that site. A model of mutagenesis is presented to explain this mutational specificity with respect to current understanding of platinum-DNA adduct structure. PMID- 8422400 TI - DNA sequencing of the seven remaining structural genes of the gene cluster encoding the energy-transducing NADH-quinone oxidoreductase of Paracoccus denitrificans. AB - In our previous papers, seven structural genes (NQO1-7) of the energy-transducing NADH-quinone (Q) oxidoreductase of Paracoccus denitrificans were characterized [Xu, X., Matsuno-Yagi, A., & Yagi, T. (1991a) Biochemistry 30, 8678-8684; (1991b) Biochemistry 30, 6422-6428; (1992a) Biochemistry 31, 6925-6932; (1992b) Arch. Biochem. Biophys. 296, 40-48]. This paper reports the identification, cloning, and sequencing of seven additional structural genes in the same gene cluster (P. denitrificans enzyme complex). These seven genes, designated NQO8-14, are composed of 1038, 492, 603, 306, 2112, 1542, and 1500 base pairs, respectively. The polypeptides encoded by the NQO8-14 genes are homologous, respectively, to the ND1 product, the 23-kDa polypeptide, and the ND6, ND4L, ND5, ND4, and ND2 products of the bovine NADH-Q oxidoreductase. The order of the 14 structural genes of the Paracoccus energy-transducing NADH-Q oxidoreductase in the gene cluster is NQ07, NQO6, NQO5, NQO2, NQO1, NQO3, NQO8, NQO9, NQO10, NQO11, NQO12, NQO13, and NQO14. Downstream from the NQO14 gene an open reading frame (designated URF240) was detected which encodes a predicted polypeptide homologous to the biotin [acetyl-CoA-carboxylase] ligase of Escherichia coli. In addition, a putative terminal sequence motif was observed downstream of the NQO14 gene, suggesting that the structural gene NQO14 is the 3'-terminal gene of the Paracoccus NADH-Q oxidoreductase gene cluster. Nucleotide sequencing of the entire gene cluster revealed the presence of three unidentified reading frames: one between the NQO3 and NQO8 genes and other two between the NQO9 and NQO10 genes. These are designated URF4, URF5, and URF6 and are composed of 768, 393, and 405 base pairs, respectively. The possible functions of the putative proteins encoded by URF5 and URF6 are discussed. PMID- 8422402 TI - Import of phosphatidylserine into isolated yeast mitochondria. AB - A yeast phosphatidylserine transfer protein was used as a tool to transport radioactively labeled phosphatidylserine from unilamellar vesicles to isolated mitochondria of the yeast, Saccharomyces cerevisiae. Import of phosphatidylserine into mitochondria was monitored by the appearance of radioactively labeled phosphatidylethanolamine, which is produced from phosphatidylserine by the action of phosphatidylserine decarboxylase. This enzyme is located on the outer aspect of the inner mitochondrial membrane. Import of phosphatidylserine into mitochondria and formation of phosphatidylethanolamine does not require ATP or an electrochemical gradient, and is not affected by adriamycin. Evidence is presented that contact sites between the two mitochondrial membranes are zones of intramitochondrial translocation of phosphatidylserine and phosphatidylethanolamine. When phosphatidylserine decarboxylase is inhibited by hydroxylamine, transport of phosphatidylserine to the mitochondrial surface is unaffected. Under these conditions only a small amount of phosphatidylserine accumulates in the inner mitochondrial membrane indicating that the intramitochondrial transport of phosphatidylserine and its metabolic conversion to phosphatidylethanolamine are linked processes. PMID- 8422403 TI - Porcine aortic endothelial cell membranes contain a LPAF: CoA-independent transacylase. AB - Membranes isolated from porcine aortic endothelial cells (PAEC) contain a CoA independent transacylase enzyme (CoA-IT). CoA-IT, an integral membrane protein, transfers an acyl moiety to added [3H]alkylhydroxyglycerophosphocholine (LPAF) to generate [3H]alkylacylglycerophosphocholine (alkylacyl-GPC). This enzyme exhibits an apparent Km of 0.7 microM and a Vmax of 0.8 nmol/min per mg for the transfer of an acyl group to added [3H]LPAF. The addition of the nonionic detergent Triton X-100 (TX-100) (0.5 mg/ml), the sulfhydryl reagents N-ethylmaleimide (NEM) (200 microM) or thimerosal (200 microM), or pre-incubating the membranes at 95 degrees C for 10 min all decreased LPAF: CoA-IT activity by more than 95%. The inhibitory action of NEM or thimerosal suggests that sulfhydryl group(s) are involved in or are close to the catalytic site of LPAF: CoA-IT. PMID- 8422404 TI - Orientation and structure of the NH2-terminal HIV-1 gp41 peptide in fused and aggregated liposomes. AB - For several retroviruses, the N-terminal hydrophobic sequence of the viral envelope glycoprotein has been shown to play a crucial role in the interaction between the virus and the host cell membrane. We report here on the interaction of a synthetic 16 residues peptide corresponding to the gp41 NH2-terminal sequence of Human Immunodeficiency Virus with the phospholipid bilayer. Fluorescence energy transfer measurements show that this peptide can induce lipid mixing of large unilamellar vesicles (LUV) of various compositions at pH 7.4 and 37 degrees C. LUV undergo fusion, provided they contained phosphatidylethanolamine (PE) in their lipid composition. To provide insight into the mechanism of the fusion event, the peptide secondary structure and orientation in the lipid bilayer were determined using Fourier Transform Infrared Spectroscopy (FTIR). The peptide adopts mainly a beta-sheet conformation in the absence of lipids. After interaction with LUV the beta-sheet is partly converted into alpha-helix. The orientation of the peptide with respect to the lipid acyl chains depends on the presence of PE in the lipid bilayer. The peptide is inserted into the lipid bilayer with the helix axis oriented parallel to the lipid acyl chains in the fused vesicles, whereas it is adsorbed parallel to the lipid/water interface in the aggregated vesicles. The role of the two kinds of orientation during the fusion event is discussed. PMID- 8422405 TI - The location of amantadine hydrochloride and free base within phospholipid multilayers: a neutron and X-ray diffraction study. AB - Concomitant neutron and X-ray studies were undertaken in order to locate accurately the anti-influenza and Parkinson's disease drug amantadine in multilayers of 1,2-dioleoyl-sn-glycero-3-phosphocholine. The X-ray data were phased using the swelling series method and the neutron data were phased using D2O/H2O exchange and a variation of the isomorphous replacement technique. The sets of data complement each other and reveal two populations of amantadine within the bilayer. One site is close to the bilayer surface, the other is much deeper. The majority of the amantadine occupies the surface site. The relative occupancy, but not the position, of the two locations appears to be dependent upon the initial protonation state of the drug. No evidence of bilayer perturbation was observed with either the protonated or the deprotonated forms of amantadine. PMID- 8422406 TI - Mutual interaction of ion uptake and membrane potential. AB - The concentration dependence of cation uptake by the cell may be considerably complicated when this uptake is accompanied by a depolarization of the cell membrane. In case of carrier-mediated transport deviations from Michaelis-Menten kinetics may come to the fore comparable to those found in a dual mechanism of cation uptake or when substrate inhibition is involved. This remains true when only the maximum rate of uptake and not the Km is dependent upon the membrane potential. We have proven this by means of computer simulation of cation transport mediated by a non-mobile carrier. Under restricted conditions still apparent Michaelis-Menten kinetics may be found despite the fact that the membrane potential varies with increasing substrate cation concentration. But even then there are still differences with 'normal' transport kinetics. A non competitive inhibitor does not only affect the maximum rate of uptake but also the apparent Km. Depolarization of the cells by a cation which passes the cell membrane by means of diffusion, affects the uptake of the substrate cation almost in the same way as a non-competitive inhibitor does and causes both a decrease in the maximum rate of uptake and an increase in Km. In the case of competitive inhibition the apparent affinity of the inhibitor for the carrier depends upon the rate of transfer of this inhibitor through the cell membrane. The mutual influence of cation uptake and membrane potential is dealt with for uniport of either monovalent or divalent cations and for cotransport of monovalent cation with protons, as well. Possible effects of the surface potential are accounted for. PMID- 8422407 TI - Properties of gel phase lipid-trehalose bilayers upon rehydration. AB - When dipalmitoylphosphatidylcholine bilayers dried under vacuum in different concentrations of trehalose are rehydrated in buffer without the sugar they show different physicochemical properties in the gel state in comparison to the normal gel state. Dry DPPC/trehalose mixtures are readily dispersed in buffer below the phase transition showing by electron microscopy a morphology similar to liposomes prepared by dispersing the lipids in buffer above the phase transition temperature. In these conditions, an increase in the peak at 570 nm of merocyanine after the dehydration-rehydration process in the presence of the sugar is observed and the water permeation increases to values comparable to those found in the fluid state as indicated by the activation energy values and the osmotic volume. The trehalose-dried liposomes rehydrated in buffer show a similar osmotic response to hypertonic gradient as DPPC liposomes without sugar near the phase transition temperature. In accordance with this behavior the trehalose-dried liposomes are lysed below the phase transition temperature by lysoderivatives. These modifications of the gel state of hydrated phospholipids by trehalose can only be achieved if a drastic dehydration is performed in the presence of the sugar. After rehydration the changes in the gel state can be detected after dyalizing the rehydrated membranes in media without trehalose during at least 24 h. These results suggest that trehalose is still intercalated between the phospholipids after restoring water to the dried liposomes either at temperatures below or above the phase transition. PMID- 8422408 TI - Transport properties and inhibitor sensitivity of isolated and reconstituted porin differ from those of intact mitochondria. AB - The pore-forming protein porin has been isolated from rat heart mitochondria and reconstituted in phospholipid vesicles of different composition. Rapid release of anions, cations and non-charged molecules has been demonstrated from the proteoliposomes but not from the protein-free liposomes. In spite of its higher molecular mass and charges, the movement of ATP was almost as fast as that of inorganic phosphate. Polyanion (1:2:3 copolymer of methacrylate/maleate/styrene), a potent inhibitor of porin residing in the mitochondrial contact sites decreased the solute movements but did not completely block any of the investigated transport processes (phosphate, chloride, ATP). Alterations of the lipid environment had significant effect: an increase in the proportion of soybean phospholipids to egg yolk phospholipids resulted in a decrease in the amount of transported substance but did not fully inhibit the ion movements. It is concluded that the transport properties of porin reconstituted in artificial phospholipid membranes are different from the characteristics of porin prevailing in the mitochondrial contact sites and additional regulatory factors are suggested to be effective in the intact mitochondria. PMID- 8422409 TI - Actin and actin-binding proteins in plasma membranes derived from Walker 256 ascites or solid tumour cells. AB - Plasma membranes from Walker 256 carcinoma cells grown ascitically or as a solid tumour were examined with respect to actin content, [3H]cytochalasin B-binding and the binding of 125I-labelled G-actin to membrane proteins separated by SDS PAGE. Differences were observed both in cytochalasin B-binding to membrane actin and affinity of 125I-labelled G-actin for specific membrane proteins. PMID- 8422410 TI - Misuse of graphical analysis in nonlinear sugar transport kinetics by Eadie Hofstee plots. AB - It has become common practice to analyse the sugar transport kinetics from initial uptake rates in Saccharomyces cerevisiae cells with Eadie-Hofstee plots. These plots often demonstrate a nonlinear behaviour. They have been resolved incorrectly into two quasilinear components indicating the presence of (at least) two uptake systems or components, with Km values differing by a factor of about 10. This graphical analysis neglects the obvious additivity of the two hypothetical systems and is therefore in error. A more efficient way to determine kinetic parameters from initial uptake experiments is to use computer-assisted nonlinear regression analysis. PMID- 8422411 TI - Chaotropic anion-phosphatidylcholine membrane interactions: an ultra high field NMR study. AB - NMR studies on the interaction of the linear chaotropic anions, SCN- and SeCN-, with sonicated egg phosphatidylcholine (EPC) vesicles have been carried out at field strengths up to 14.1 Tesla. At 600 MHz, both anions cause splitting or increased splitting of the choline N+(CH3)3, CH2N+ and O3POCH2 1H resonances with SeCN- being somewhat more effective in this action than is SCN-. No changes were observed in the glycerol CH2OP and CH2OCO 1H resonances and the phosphate 31P resonance of the headgroup region. The 13C spectrum was unchanged by the presence of the anions. After 18 h of exposure to the anion, the 1H resonance splittings but not the chemical shift values returned to those prior to anion exposure. Increasing the temperature of the vesicles decreased the anion-induced splitting, but, upon return to the beginning temperature, the chemical shifts did not return to their original values. The results are considered in terms of the 'molecular electrometer' model recently developed by Seelig and co-workers [1]. PMID- 8422412 TI - Assembly and channel-forming activity of a naturally-occurring nicked molecule of Staphylococcus aureus alpha-toxin. AB - From the culture supernatant of Staphylococcus aureus Wood 46, we obtained a naturally-occurring nicked molecule of staphylococcal alpha-toxin. The nicked alpha-toxin consisted of non-covalently-linked 8-kDa and 25-kDa polypeptides, which were derived, respectively, from the N-terminal and the C-terminal part of the toxin nicked at the peptide bond between Glu-71 and Gly-72. The nicked toxin, as well as native alpha-toxin, bound to and oligomerized in the liposome membranes composed of choline-containing phospholipids (i.e., phosphatidylcholine and sphingomyelin) and cholesterol, and formed membrane channel in the liposome membranes. However, the channel-forming activity of the nicked toxin, assessed as a toxin-induced carboxyfluorescein leakage from the liposomes, was approx. 20 fold lower than that of native alpha-toxin. Channel-forming activity of the nicked toxin as well as native toxin was inhibited by divalent cations including Zn2+, Cd2+, Ca2+ and Mg2+, and degree of the inhibitory effect of the divalent cations was in the following order: Zn2+ > Cd2+ > Ca2+ > Mg2+. Thus, although the cleavage of alpha-toxin at the position between Glu-71 and Gly-72 drastically reduced channel-forming activity of the toxin, the nicked toxin retained the ability to oligomerize in phospholipid-cholesterol membranes and the characteristics of channel-forming activity in terms of the specificity for phospholipids and the susceptibility to divalent cations. PMID- 8422413 TI - Effect of sub-skinning concentrations of saponin on intracellular Ca2+ and plasma membrane fluidity in cultured cardiac cells. AB - To determine the underlying mechanisms of the positive inotropic effect of sub skinning concentrations of saponin, we studied changes in the intracellular Ca2+ ([Ca2+]i) and plasma membrane fluidity after exposure to digitonin (a representative saponin) in cultured cardiac cells. [Ca2+]i was measured by use of the fluorescent calcium indicator Calcium Green-1. The membrane fluidity was evaluated by measuring the diffusion coefficient using the method of fluorescence recovery after photobleaching. 1,1'-Dioctadecyl-3,3,3',3' tetramethylindodicarbocyanine perchlorate was used as the fluorescent probe. Digitonin at a sub-skinning concentration (0.1 to 1 microM) produced an increase in cell motion and an augmentation of [Ca2+]i. Membrane fluidity which is evaluated by the diffusion coefficient (from 0.34.10(-8) to 0.28.10(-8) cm2/s; P < 0.05), decreased in the presence of 0.2 microM digitonin while the cell maintained an augmented motion and an increased [Ca2+]i. The skinning concentration of digitonin (5 microM) produced a rapid contracture with a marked increase in [Ca2+]i. The membrane fluidity was further reduced (diffusion coefficient: 0.24.10(-8) cm2/s; P < 0.001). These results suggest that saponin at the sub-skinning concentration also causes holes in the plasma membrane by interaction with cholesterol, as was shown with the skinning concentration, and it increases [Ca2+]i, which thereby induces a positive inotropic effect. PMID- 8422414 TI - Structural and functional properties of rhodopsin from rod outer segment disk and plasma membrane. AB - The structural and functional properties of bovine rhodopsin from rod outer segment disk and plasma membranes were compared by high performance liquid chromatography (HPLC), mass spectrometric analyses, and in vitro rhodopsin phosphorylation assays. Disk and plasma membranes separated by a ricin gold dextran affinity perturbation method were treated with trypsin or cyanogen bromide, and the N-terminal and C-terminal rhodopsin peptides were isolated by immunoaffinity chromatography using antirhodopsin monoclonal antibodies coupled to Sepharose. Reverse phase HPLC chromatograms of the C-terminal and N-terminal peptides from disk and plasma membrane rhodopsin were found to be similar. Mass spectrometric, PicoTag, and hexose analyses of the tryptic 1-16 N-terminal peptides further indicated that the post-translational glycosylation of plasma membrane rhodopsin is identical to that of disk membrane rhodopsin. HPLC analysis of soluble peptides obtained from cyanogen bromide and tryptic digestion of immunoaffinity purified rhodopsin also indicated that no significant differences exist between disk and plasma membrane rhodopsin. Light-induced phosphorylation of rhodopsin in disk and plasma membranes were also compared using in vitro phosphorylation assays. Plasma membrane rhodopsin was found to undergo light dependent, rhodopsin kinase catalyzed phosphorylation to the same extent as disk membrane rhodopsin. These results indicate that the bulk rhodopsin in rod outer segment plasma membranes appears to be identical to rhodopsin in disk membranes in regard to primary structure, post-translational glycosylation and light dependent phosphorylation. On this basis, it is unlikely that the sorting of rhodopsin between disk and plasma membranes occurs by a mechanism based on differences in structural properties of rhodopsin. PMID- 8422415 TI - Dehydration-induced lamellar-to-hexagonal-II phase transitions in DOPE/DOPC mixtures. AB - Plasma membranes of protoplasts isolated from non-acclimated rye plants undergo a transition from the bilayer to the inverted hexagonal (HII) phase during freeze induced dehydration at -10 degrees C. It has been suggested (Bryant, G. and Wolfe, J. (1989) Eur. Biophys. J. 16, 369-372) that the differential hydration of various membrane components may induce fluid-fluid demixing of highly hydrated (e.g., PC) from poorly hydrated (PE) components during dehydration. This could yield a PE-enriched domain more prone to form the HII phase. We have examined the lyotropic phase behavior of mixtures of DOPE and DOPC at 20 degrees C by freeze fracture electron microscopy, differential scanning calorimetry, and X-ray diffraction. HII phase formation was favored by higher proportions of DOPE and lower water contents. Mixtures of 1:1 and 1:3 DOPE/DOPC had a hydration-dependent appearance of two L alpha phases at water contents just above those at which the HII phase occurred. The hydration-dependence of the lamellar repeat spacings suggested that the DOPE-enriched domains preferentially underwent the L alpha-to HII phase transition. Mixtures of 3:1 DOPE/DOPC did not separate into two L alpha phases during dehydration. These data suggest that the differential hydration characteristics of various membrane components may induce their lateral fluid fluid demixing during dehydration. PMID- 8422416 TI - Purification and properties of an oxalate oxidase from leaves of grain sorghum hybrid CSH-5. AB - An oxalate oxidase (EC 1.2.3.4), which catalyzes aerobic oxidation of oxalate to CO2 and H2O2, has been purified to apparent homogeneity from 10-day-old leaves of grain sorghum hybrid CSH-5, as determined by disc-gel electrophoresis. The molecular weight of the enzyme was about 120,200 by Sephadex G-200 gel filtration, 62,000 by SDS disc-gel electrophoresis. The enzyme had an optimum pH of 5.0 and activation energy of 4.4 kcal/mol. The rate of reaction was linear up to 2 min. The Km value for oxalate was 0.78.10(-4) M. The enzyme was inhibited by EDTA, L-cysteine and sodium azide, but iodoacetate had no effect. The enzyme was strongly stimulated by Cu2+ and was unaffected by chloride ions in physiological concentration range. The better suitability of the enzyme for urinary oxalate determination is demonstrated. PMID- 8422417 TI - Characterization of the activation of pro-urokinase by thermolysin. AB - The bacterial metalloproteinase thermolysin catalyzes the efficient activation of pro-urokinase to an active high-molecular-weight form of the protein. Thermolysin and plasmin convert pro-urokinase to enzymes of essentially equal activities in amidolytic assays, but with different molecular structures. The B-chains of the proteins produced by thermolysin and plasmin are of the same size (33 kDa) and have the same amino-terminal sequences, demonstrating that the cleavage of the Lys158-Ile159 bond of pro-urokinase is catalyzed by both enzymes. However, thermolysin also reacts at additional sites in the growth factor domain of the A chain at nearly the same rate as that of the activation reaction. Polypeptides derived from hydrolyses of the Glu3-Leu4, Tyr24-Phe25, Asn27-Ile28 and Lys36 Phe37 bonds are recovered after reduction of the activated protein. The carboxy terminus of the A-chain has been shown to be Arg-156, a consequence of proteolysis of the Arg156-Phe157 bond. In contrast to plasmin, thermolysin activates thrombin-inactivated pro-urokinase nearly as rapidly as it does the native zymogen. Thermolysin provides a useful alternative to plasmin for the catalytic activation and analysis of pro-urokinase, since the bacterial metalloproteinase is stable in solution and not susceptible to inhibition by aprotinin and other serine proteinase inhibitors. PMID- 8422418 TI - Transition-state discrimination by adenosine deaminase from Aspergillus oryzae. AB - Adenosine deaminase from Aspergillus oryzae resembles mammalian adenosine deaminases in its ability to catalyze the hydrolytic removal of many substituents from C-6, and in the chirality at C-6 of the active isomer of the transition state-analogue inhibitor 6-hydroxymethyl-1,6-dihydropurine ribonucleoside. The 5' OH group of adenosine has been found to contribute a factor of 5.10(4) to transition-state stabilization by calf intestinal adenosine deaminase, and crystallographic observations suggest that a zinc-histidine 'bridge' is formed between the 6-OH and the 5'-OH groups of the substrate in the transition state for its deamination. The present paper describes experiments indicating that this bridge is not present during the action of adenosine deaminase from Aspergillus oryzae. We find (1), that the fungal enzyme catalyzes deamination of adenosine and 5'-deoxyadenosine with kcat/Km values that are almost identical; (2), that the Ki value of the transition-state-analogue inhibitor 2'-deoxycoformycin is much higher for the fungal enzyme (2.7.10(-9) M) than for the mammalian enzyme (2.10(-12) M) and (3), that this difference in binding affinities arises mainly from a difference in rates of enzyme-inhibitor association. Thus, the onset of inhibition was markedly slower for the fungal enzyme (kon = 1.3.10(4) M-1 s-1) than for the calf intestinal enzyme (kon = 2.6.10(6) M-1 s-1). Effects of chelating agents and divalent cations suggest that the fungal enzyme, like other deaminases for adenosine and cytidine, contains essential zinc. PMID- 8422419 TI - H-NMR characterization of the human myocardium myoglobin and erythrocyte hemoglobin signals. AB - The 1H-NMR signal of deoxy Mb provides a unique opportunity to measure tissue oxygenation in vivo. To utilize the technique for human application, however, requires a specific spectral characterization of both human Mb and erythrocyte Hb. We report that the proximal histidyl-NH signal of human deoxy Mb resonates at 80.3 ppm at 25 degrees C and maintains a 3.9 ppm separation with the corresponding Hb A signal throughout the physiological temperature range. In the particular case of the human thenar muscle, the deoxy Mb signal is clearly detectable without any interference from Hb. PMID- 8422420 TI - Identification of the sites of modification on bovine carbonic anhydrase II (BCA II) by the salt-bridge reagent cyanogen, C2N2. AB - The hydrolase activities of bovine carbonic anhydrase B (BCA II carbonate hydrolyase, EC 4.2.1.1) were modified by cyanogen (C2N2, N identical to C-C identical to N, ethanedinitrile) with decreases in Vmax of as much as 99%. This was not accompanied by a reduction in hydrolyase activity. These changes were not reversed at lower pH values but the enzymatic activity was restored by incubation at pH 10. 14C-labeled glycine ethyl ester ([14C]GEE) specifically and covalently bound to the cyanogen-treated BCA II, as verified by HPLC and 14C monitoring. It was shown that sites of cyanogen-introduced modifications in BCA II could be effectively labeled and identified by incubation with the nucleophile [14C]GEE. Three radiolabeled tryptic peptides from BCA II arising from a labeling process designed to study cyanogen-induced modifications leading to nucleophile labile covalent bonds have been isolated. The residues identified by [14C]GEE labeling were Asp-34, Glu-117 and Asp-152. Three moieties attached to the omega-carboxyls by C2N2 were tentatively identified by molecular modeling; they were Arg-111, His 107 and/or His-119 and Ser-216, respectively. The use of C2N2 afforded a means to compare the salt bridges in two species and showed that two of three were not conserved. PMID- 8422421 TI - Influence of the N- and C-terminal chains on the zinc-binding and conformational properties of the central zinc-finger structure of Moloney murine leukaemia virus nucleocapsid protein: a steady-state and time-resolved fluorescence study. AB - The nucleocapsid protein NCp10 of the Moloney murine leukaemia virus is a small basic protein characterized by a central Cys26-X2-Cys29-X4-His34-X4-Cys39 zinc finger domain. Mutants with deletion of either the N- or C-terminal chain (or both) surrounding the central zinc-finger domain were synthesized by a solid phase approach in order to evaluate the influence of these lateral chains on zinc binding and conformational properties of NCp10. For this purpose, the steady state and time-resolved fluorescence properties of the single Trp-35 residue of the various NCp10 derivatives were analyzed. The binding properties of the various derivatives suggest that the central zinc-finger domain affinity for zinc is not modified by the N-terminal chain and is only slightly (about one order of magnitude) increased by the C-terminal chain leading to a Kapp of (1.2 +/- 0.2).10(14) M-1 for the whole NCp10. Concerning the conformation of the NCp10 derivatives, fluorescence data are in agreement with structureless polypeptide chains in the absence of zinc. In contrast, in the presence of zinc, the fluorescence intensity decays are in agreement with a unique conformation of the finger motif backbone and a distribution of the Trp-indole moiety into two classes with different local environments. Decay-associated spectra, fluorescence quenching by acrylamide and anisotropy decay data further suggest that the Trp indole moiety of both classes was highly exposed to solvent and had a high degree of rotational freedom. Finally, in contrast to the C-terminal chain, the N terminal chain modifies the local environment and the accessibility to external quenchers of both Trp-35 classes, suggesting that it was folded in the vicinity of the Trp-35 residue. PMID- 8422422 TI - Effect of salt concentration of buffer on the binding of sodium dodecyl sulfate and on the viscosity behavior of the protein polypeptide derived from bovine serum albumin in the presence of the surfactant. AB - The complex between SDS and a protein polypeptide derived from bovine serum albumin was characterized with respect to binding of SDS and viscosity behavior. The amount of bound SDS increased from 1.0 to 2.2 g/g with increase of the buffer concentration from 10 to 220 mM. A logarithmic plot of the amount of bound SDS against the buffer concentration gave a linear relation like in the plot where the number of SDS molecules constituting a spherical micelle of SDS is plotted similarly. The increase in the buffer concentration up to 25 mM, from 25 to 100 mM and beyond 100 mM, was accompanied by a sharp rise, monotonic decrease and levelling-off of the intrinsic viscosity in the respective region. In the region 45-175 mM, a linear relation was found between the intrinsic viscosity and reciprocal square root of the buffer concentration. The observed changes can be interpreted as follows: (1), the electrostatic repulsion between charges introduced by the bound SDS caused the initial increase; (2), shielding of the charges as the result of ion condensation with further increase in ionic strength caused the viscosity drop and subsequent levelling-off. The characteristics of the plots are consistent with the necklace model proposed previously for such complexes in which SDS is bound to a protein polypeptide forming micelle-like clusters and which behave like a flexible polyelectrolyte (Shirahama, K., Tsujii, K. and Takagi, T. (1974) J. Biochem. 75, 309-319). PMID- 8422423 TI - The kinetic mechanism of yeast phosphoenolpyruvate carboxykinase. AB - The kinetic mechanism of yeast phosphoenolpyruvate carboxykinase, in the physiological direction, has been determined. Product inhibition using KHCO3 showed competitive inhibition, when both oxalacetate (OAA) and ATP were varied. Phosphoenolpyruvate showed noncompetitive inhibition against OAA, and competitive inhibition with respect to ATP. Conversely, ADP showed competitive inhibition against OAA and noncompetitive inhibition vs. ATP. Dead-end inhibition studies with beta-sulfopyruvate showed competitive inhibition against OAA and noncompetitive inhibition vs. ATP. Ethene-ATP exhibited competitive inhibition against ATP and noncompetitive inhibition with respect to OAA. These results are consistent with a random Bi-Ter mechanism with the formation of two abortive complexes: enzyme-ATP-ADP and enzyme-OAA-PEP. PMID- 8422424 TI - Structure and composition of ferritin cores from pea seed (Pisum sativum). AB - Iron cores from native pea seed (Pisum sativum) ferritin have been analysed by electron microscopy and Mossbauer spectroscopy and shown to be amorphous. This correlates with their relatively high phosphate content (Fe: P = 2.83; 1800 Fe, 640 P atoms/molecule). Reconstituted cores obtained by adding iron (2000 Fe atoms/molecule) in the absence of phosphate to pea seed apoferritin were crystalline ferrihydrite. In vitro rates of formation of pea-seed ferritin iron cores were intermediate between those of recombinant human H-chain and horse spleen apoferritin and this may reflect the amino-acid residues of its ferroxidase and putative nucleation centres. The high phosphate content of pea seed ferritin suggests that this molecule could be involved in both phosphorus and iron storage. The high phosphate concentration found within plastids, from which the molecules were isolated, is a possible source of the ferritin phosphate. PMID- 8422425 TI - Purification and characterization of two metalloproteinases from squid mantle muscle, myosinase I and myosinase II. AB - Metalloproteinases, myosinase I and myosinase II, that hydrolyze the heavy chain of myosin, were purified from squid mantle muscle. Myosinase I does not hydrolyze other muscle proteins, casein, haemoglobin, or MCA-substrates, while II hydrolyzes tropomyosin. Both myosinase I and myosinase II gave a single protein band on SDS-PAGE with a molecular mass of 16 and 20 kDa, respectively. Their activities were inhibited by EDTA and 1,10-phenanthroline, and II was also inhibited by EGTA. They could be reactivated with some divalent cations, I was especially reactivated with Co2+ and II especially with Zn2+. The optimum pH of both activities was 7.0; the optimum temperature for both was 40 degrees C. Myosinase I hydrolyzes myosin heavy chains to produce 130 and 90 kDa fragments. The N-terminal amino-acid sequence of the 90 kDa fragment indicates that myosinase I splits the myosin heavy chain between Ala-1161 and Thr-1162 in subfragment 2. Myosinase II hydrolyzes myosin heavy chain to produce 158 and 65 kDa fragments, and it splits between Glu-1381 and Thr-1382 in LMM. Myosinases I and II are most likely related to the metabolism of myosin in vivo. PMID- 8422426 TI - The effect of heparin on cell proliferation and type-I collagen synthesis by adult human dermal fibroblasts. AB - (1), A commercial heparin inhibited the proliferation of two normal human dermal fibroblast cultures in a dose-responsive manner. 300 micrograms heparin/ml gave the maximum inhibition of 65%. Proliferation of fibroblasts from a patient with progressive systemic sclerosis was similarly inhibited by heparin. Heparan sulphate, pentosan polysulphate and a fucoidan also inhibited proliferation of the fibroblast cultures but chondroitin sulphate, dermatan sulphate and hyaluronate had no effect. (2) Type-I collagen synthesis per cell was elevated by up to 1.5-fold in the normal fibroblast cultures when proliferation was inhibited by heparin, heparan sulphate, pentosan polysulphate and the fucoidan. Progressive systemic sclerosis fibroblasts synthesized more collagen than the normal cell cultures and this was further stimulated by heparin and pentosan polysulphate. In contrast, heparin and the other polysulphates inhibited type-I collagen synthesis by about 20-30% in normal confluent fibroblast cultures. Collagen synthesis by confluent systemic sclerosis fibroblasts was reduced by only about 10%. (3), Total [35S]proteoglycan synthesis per cell was greatly elevated (approx. 2.5 fold) in normal subconfluent cultures treated with heparin. In contrast, heparin stimulated only a small increase in [35S]sulphate incorporation into proteoglycans in confluent cultures. PMID- 8422427 TI - Synergistic interactions of etoposide and interleukin-1 alpha are not due to DNA damage in human melanoma cells. AB - Several possible mechanisms of the synergistic interactions of IL-1 alpha and VP 16 against A375-C6 human melanoma cells were investigated. Studies indicate that IL-1 alpha did not increase topoisomerase II-dependent VP-16-mediated DNA damage, nor did IL-1 alpha inhibit the repair of VP-16-induced DNA damage in these cells. Furthermore, IL-1 alpha by itself or in combination with VP-16 did not cause significant fragmentation of cellular DNA into oligomers, indicating programmed cell death (apoptosis) was not involved in the mechanism of synergy. In contrast, an IL-1-specific receptor antagonist significantly decreased IL-1 alpha toxicity toward the melanoma cells and nearly eliminated the synergistic interactions of IL-1 alpha with VP-16. These results strongly indicate that synergism of IL-1 alpha with VP-16 was dependent upon an IL-1-receptor-mediated processes. DNA strand breakage was unlikely to be a primary intracellular target for IL-1 alpha cytotoxicity and synergism with VP-16. PMID- 8422428 TI - Increased expression of the mRNA for hormone-sensitive lipase in adipose tissue of cancer patients. AB - The expression of genes coding for regulatory enzymes involved in the uptake, synthesis and mobilisation of lipid was measured in adipose tissue of cancer patients. Total RNA was isolated from subcutaneous adipose tissue of control and cancer patients and the various mRNAs measured by Northern blot analysis. The total lipoprotein lipase enzymic activity and the relative levels of the mRNAs for lipoprotein lipase and for fatty acid synthase were not significantly different between cancer patients and control patients. However, there was a significant two-fold increase in the relative level of mRNA for hormone-sensitive lipase (HSL) in adipose tissue of cancer patients compared with control patients. The cancer patients also exhibited a two-fold elevation in serum triacylglycerol levels and serum free fatty acid levels. There was a significant correlation between the serum free fatty acid level and expression of HSL mRNA in the adipose tissue. The serum levels of insulin and tumour necrosis factor-alpha were not different between cancer and control patients. The results suggest that at least one of the mechanisms for depletion of lipid from adipose tissue in cancer patients operates at the level of increased expression of mRNA of the lipolytic regulatory enzyme, hormone-sensitive lipase. PMID- 8422429 TI - Decreased messenger RNA of arachidonate 12-lipoxygenase in platelets of patients with myeloproliferative disorders. AB - On the basis of the previous work by Okuma and Uchino [Blood 54, 1258-1271, 1979], three patients with myeloproliferative disorders were investigated with a special reference to arachidonate 12-lipoxygenase in their platelets. The cytosol of the patients' platelets showed a markedly reduced activity of arachidonic acid oxygenation to 12-hydroperoxy acid. A peroxidase-linked immunoassay for the 12 lipoxygenase demonstrated only 7-12% of the normal level of the enzyme protein in the cytosol fraction of platelets. Furthermore, 12-lipoxygenase mRNA level was determined quantitatively by a reverse transcriptase-polymerase chain reaction with an internal standard cRNA which was synthesized by in vitro transcription of human platelet 12-lipoxygenase cDNA with a 105-bp deletion. The 12-lipoxygenase mRNA content was 4.7 +/- 2.0 (mean +/- S.D.) ng/10(11) platelets in 13 normal subjects. In contrast, the mRNA content was as low as 0.15, 0.11 and 0.10 ng/10(11) platelets in the three patients. Taken together, the 12-lipoxygenase deficiency in these patients was attributable to the decreased 12-lipoxygenase mRNA level and thus the impaired synthesis of the enzyme protein in their platelets. PMID- 8422430 TI - Localization of cells in the rat brain expressing fumarylacetoacetate hydrolase, the deficient enzyme in hereditary tyrosinemia type 1. AB - Fumarylacetoacetate hydrolase (FAH) is the terminal enzyme in the catabolic pathway of tyrosine. This enzyme which is mainly expressed in the liver and kidney is deficient in hereditary tyrosinemia type 1. As some affected individuals present neurologic abnormalities, we studied the expression of FAH in the rat and human brain. The FAH gene was shown to be expressed in the rat brain by immunoblot and Northern blot analysis. The FAH protein was also detected in human brain by the immunoblot assay. An immunohistochemical study was undertaken to localize the FAH-producing cells in the rat central nervous system. This analysis showed that the majority of FAH-producing cells are localized in the axonal nerve fibers of the white matter, although positive cells could also be found throughout the brain. The greatest number of FAH-positive cells were found in structures consisting essentially of white matter, such as the corpus callosum. This specific localization in the white matter indicates that some type of glial cells are responsible for the expression of the FAH protein in the rat central nervous system. The characteristic linear organization found in some of the FAH-positive cells in the corpus callosum suggests that these glial cells are oligodendrocytes. These findings are discussed with respect to the neurologic symptoms observed in some tyrosinemia patients. PMID- 8422431 TI - Immunoblot analysis of dystrophin-related protein (DRP). AB - Polyclonal antibodies against the carboxy-terminal portion of dystrophin-related protein (DRP), the putative autosomal gene product which shares sequence homology with dystrophin, show the clear expression of DRP in mouse fetal muscle and in cultured human muscle cells, but not in mature mouse or human muscle. DRP has the same molecular mass as X-linked dystrophin and is recovered from the membrane fraction, but is associated with membranes more loosely than dystrophin. PMID- 8422432 TI - Incorporation of [14C]hypoxanthine into cardiac adenine nucleotides: effect of aging and post-ischemic reperfusion. AB - In order to investigate whether the 'hypoxanthine salvage' pathway of the cardiac muscle is modified with age, we aerobically perfused isolated hearts of 4-month- and 22-month-old male Wistar rats for 20 min with 0.18 microM [14C]hypoxanthine. A second group of hearts was subjected to a 30-min ischemic perfusion (95% reduction of the coronary flow), followed by 20 min of reperfusion. In this last 20 min, the perfusate contained the same concentration of [14C]hypoxanthine used under the aerobic condition. After 20 min of aerobic perfusion the myocardial levels of ATP were significantly lower (15%) in aged than young rat hearts, whilst no age-related differences were observed at the end of the reperfusion. In the young rats the incorporation of the isotope into ATP, ADP, and AMP was significantly higher (192%, 226%, and 300%, respectively), after 20 min of reperfusion with respect to the aerobic values. On the contrary, in the aged hearts, no significant change in the rate of [14C]-incorporation into ATP was observed during reperfusion, despite an increase of the [14C]-incorporation into ADP and AMP. Moreover, the content of each labeled adenine nucleotide was significantly higher in aged than young hearts at the end of the aerobic period, whereas the incorporation of the labeled hypoxanthine was not affected by age after 20 min of reperfusion. The release of uric acid into coronary effluents was greater (50%) in aged than young rats during the reperfusion period, but no age dependent differences in the isotope incorporation into uric acid were observed. These data indicate that in the aged rat heart, perfused under aerobic conditions, there is an increased incorporation of hypoxanthine into ATP, although it does not further increase during postischemic reperfusion. PMID- 8422433 TI - Difference between human and guinea pig Hageman factors in activation by bacterial proteinases: cleavage site shift due to local amino acid substitutions may determine the activation efficiency of serine proteinase zymogens. AB - Human and guinea pig Hageman factors have been subjected to the action of pseudomonal elastase and serratial E15 proteinase. The pseudomonal elastase cleaved 22-24% of the human molecule at Arg353-Val354, and the remainder at Gly357-Leu358 resulting in the generation of about 20% of potential activity as activated Hageman factor, compared with trypsin activation, while it hydrolyzed Arg340-Ile341 bond in guinea pig molecule and generated about 75% of activity as activated Hageman factor. The serratial proteinase did not hydrolyze the essential cleavage site (Arg353-Val354) of the human zymogen but Gly356-Gly357 (30%) and Gly357-Leu358 (70%) bonds. Both products showed no activity. The guinea pig zymogen, in contrast, was cleaved mostly at Arg340-Ile341 (70%) and less abundantly at Gly344-Leu345 (30%), generating about 85% of the whole potential activity as activated Hageman factor. From the high correspondence between the proportions of activation and of hydrolysis at the essential cleavage site in activation, it was concluded that hydrolysis of the bonds different from the essential bond did not cause activation, even when the spatial separation was only 3 or 4 residues. Considering the amino acid differences between human and guinea pig Hageman factors, -Met351-Thr-Arg-Val-Val-Gly-Gly-Leu-Val-Ala360- and Leu338-Ser-Arg-Ile-Val-Gly-Gly-Leu-Val-Ala347-, respectively, it was realized that even the minor amino acid substitutions caused the cleavage site shift which resulted in significant differences in activation efficiency of the proteinase zymogens. PMID- 8422434 TI - A biochemical basis for synergism of 6-mercaptopurine and mycophenolic acid in Molt F4, a human malignant T-lymphoblastic cell line. AB - 6-Mercaptopurine (6MP) cytotoxicity was studied in Molt F4 cells, a T-cell acute lymphoblastic leukemia (ALL) cell line. The effects on cytotoxicity were concentration-dependent. Measurements of intracellular thionucleotide intermediates of 6MP demonstrated a rapid rise of thio-IMP (tIMP) levels, and subsequently a rapid decrease. Thio-GMP (tGMP) and methyl-thio-IMP (Me-tIMP) appeared later in time, and persisted longer. Mycophenolic acid (MPA), a specific inhibitor of IMP dehydrogenase (IMPDH), was used to inhibit the conversion of tIMP into tGMP, thereby decreasing the incorporation of 6MP into DNA. A synergistic effect on cell viability and cell growth was observed when cells were treated with a combination of 2 microM 6MP and 0.5 microM MPA. Also, intracellular Me-tIMP increased 5 times with the combination. Based on the increase of Me-tIMP concentration and the observed synergism between 6MP and MPA, we conclude that methylation of tIMP into Me-tIMP is an important alternative route for 6MP cytotoxicity. PMID- 8422435 TI - Divergent effects of butyrate on the alkaline phosphatases of SW-620 cells. AB - The human colon cancer cell line SW-620 produces two alkaline phosphatases (ALPs) which are not expressed by normal colon. They are the heat-stable, term-placental and the heat-labile, L-homoarginine-sensitive, liver/bone/kidney ALPs. Butyrate, an ALP inducer, has strikingly dissimilar effects on the activity of these enzymes: whereas high (2.0 mM) butyrate concentrations exclusively induce increased activity levels of liver/bone/kidney ALP, low (0.5 mM) concentrations increase the activity of both, albeit induction of term-placental ALP is less pronounced. These observations indicate that the effect of butyrate on the two ALPs is non-coordinate and suggest that their expression by SW-620 cells is independently modulated. PMID- 8422436 TI - Specific non-enzymatic glycation of the rat histone H1 nucleotide binding site in vitro in the presence of AlF4-. A putative mechanism for impaired chromatin function. AB - We show here that an aluminium derivative, AlF4-, stimulates glycation of histone H1 selectively in the proximity of its nucleotide-binding site. This adduct formation interferes with nucleoside triphosphate hydrolysis by H1 and with nucleotide modulation of H1 DNA binding. The present mode of aluminium action may in part be responsible for its effects on the chromatin structure and expression of tissue-specific genes, and may constitute a mechanism in the pathogenesis of aluminium-induced encephalopathy and in that of Alzheimer's disease, for example. PMID- 8422437 TI - The interaction of soluble human complement receptor type 1 (sCR1, BRL55730) with human complement component C4. AB - Human CR1 is a membrane-bound protein which plays an important role in the control of the human complement system. In addition to its involvement in the processing and clearance of immune complexes with C3b or C4b on their surface, CR1 acts as a cofactor for the proteolysis of C3b and C4b by Factor I. sCR1 is a recombinant, soluble form of CR1 which retains the cofactor activities of CR1, and is of potential therapeutic value for the suppression of complement-mediated tissue damage in vivo. An assay has been established using microtitre plates to explore the binding of sCR1 to the two isotypes of C4, C4A and C4B, and to C4 fragments. Specific binding of 125I-sCR1 to C4b and ammonia-treated C4 has been demonstrated. The binding of 125I-sCR1 to ammonia-treated C4 is dependent on pH and ionic strength, decreasing with an increase in pH and with an increase in ionic strength. At physiological ionic strength, up to twice as much 125I-sCR1 bound to ammonia-treated C4A as bound to ammonia-treated C4B. This preference of sCR1 for binding to the C4A isotype has implications for the clinical association of immune complex disease with C4A null alleles. PMID- 8422438 TI - Chronic hyperammonemia prevents changes in brain energy and ammonia metabolites induced by acute ammonium intoxication. AB - Acute ammonia toxicity has been attributed to the depletion of energy metabolite intermediates. Ingestion of an ammonium containing diet produces hyperammonemia and protects rats against acute ammonium intoxication. We have tested the effect of chronic hyperammonemia on the brain contents of energy and ammonia metabolite intermediates and on the effect on these contents of acute ammonia intoxication (i.p. injection of 7 mmol/kg of ammonium acetate). Chronic hyperammonemia was induced in rats by feeding them a diet containing 20% ammonium acetate. Control rat were fed the same diet without addition of ammonium acetate. It is shown that chronic hyperammonemia did not affect the content of most metabolites, the only remarkable changes are the increases of the contents of ammonia (46%), glutamine (81%), acetoacetate (31%) and of the mitochondrial NAD+/NADH ratio (32%) as well as the marked decrease of beta-hydroxybutyrate (by 86%). Chronic hyperammonemia prevents most changes in metabolites induced by acute ammonium intoxication (i.p. injection of 7 mmol/kg of ammonium acetate). In control rats it was a marked breakdown of glycogen and increased contents of glucose, lactate and pyruvate, with decreased cytosolic NAD+/NADH ratio and beta-hydroxybutyrate and ATP contents. These changes were nearly completely prevented in hyperammonemic rats. In controls, ammonia increased 12.8-fold while glutamate and aspartate decreased by approximately 40% and glutamine and alanine raised by 37% and 93%, respectively; in hyperammonemic rats ammonia increased 6.9-fold while glutamate, glutamine and alanine were not significantly affected. Also the mitochondrial NAD+/NADH ratio raised by 18-fold in controls and by 6-fold in hyperammonemic rats. These results indicate that chronic hyperammonemia markedly prevents the alterations of the contents of energy and ammonia metabolites induced by acute ammonium intoxication. PMID- 8422439 TI - Very long chain acyl-CoA dehydrogenase deficiency: identification of a new inborn error of mitochondrial fatty acid oxidation in fibroblasts. AB - A patient highly suspected of long chain fatty acid oxidation defect was investigated for membrane-bound palmitoyl-CoA dehydrogenation in a membrane extract from skin fibroblasts, using 1% sodium cholate as detergent. The profoundly decreased activity observed is consistent with a deficiency of the newly identified mitochondrial 'very long chain acyl-CoA dehydrogenase'. PMID- 8422440 TI - The role of alcohol in work-related fatal accidents in Australia 1982-1984. AB - This paper describes the role of detectable blood alcohol in fatal work injuries. An attempt was made to identify all work-related fatalities that occurred throughout Australia in the period 1982-1984. A research team examined coroners' records and classified 1737 fatal injury cases as being work-related according to study definitions. The following were also extracted from data in coroners' records: (i) whether or not the description of the fatal event indicated that inebriation was likely to have been a factor; (ii) whether or not there was documentation of blood alcohol concentration (BAC), and if so, what it was. The likelihood of inebriation was assessed without knowledge of the victim's BAC. In 1030 (59 per cent) of the 1737 fatal work injury cases, a BAC determination was documented. Zero levels were detected in 867 fatalities (84 per cent), and 163 cases (16 per cent) had non-zero BAC. In the latter group the median BAC was 104 mg%. Sixty-five per cent of measurable BAC cases had BAC greater than 50 mg%. Fatality risk in the non-zero BAC group relative to that of the zero BAC group was elevated for the following factors: marital status--single (risk ratio (RR) = 1.7, 95 per cent confidence interval (CI) 1.1-2.8) or separated/divorced (RR = 2.4, CI 1.5-3.8); occupation as manager, executive or administrator (RR = 2.5, CI 1.5-5.8); and commuting (RR = 1.6, CI 1.2-2.0).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422441 TI - Exposure to neurotoxic metals among workers in Singapore: an overview. AB - The extent of occupational exposure to inorganic lead, manganese, arsenic and inorganic mercury in Singapore was determined from the results of Statutory Medical Examinations and environmental monitoring carried out by the Department of Industrial Health in 1989. There were 786 workers exposed to lead. Of these, 7.8 per cent had blood levels greater than 40 micrograms/dl. There were 67 workers exposed to mercury, 11.9 per cent of whom had urinary mercury levels greater than 50 micrograms/l. There were 101 and 144 workers exposed to arsenic and manganese respectively. None of the biological samples exceeded the health based limits. A review of local studies showed that some of the exposed workers had neurophysiological and neurobehavioural changes. PMID- 8422442 TI - Audit of near patient cholesterol testing in occupational health clinics. AB - An audit of near patient cholesterol testing was carried out in occupational health clinics. The aims were to examine the statistical agreement between Reflotron and laboratory measurements of blood cholesterol and to formulate a policy for the use of Reflotrons in cholesterol testing. Three hundred and fifty two staff members attending occupational health clinics over a period of 10 months had blood taken for both Reflotron and laboratory measurements. The correlation between the two methods was 0.95. The Reflotron had a negative bias compared to the laboratory, with the mean difference between the two methods of measurement being -0.21 mmol/l (95 per cent confidence interval -0.18 to -0.25 mmol/l). Despite the high correlation coefficient and small mean difference, the scatter of Reflotron-laboratory differences was broad, with 95 per cent of the differences lying in the range of 0.95 mmol/l below to 0.52 mmol/l above the laboratory result. For Reflotron results of 5.50 mmol/l and greater, the sensitivity and specificity of the Reflotron in detecting subjects with laboratory cholesterol levels greater than 6.5 mmol/l were 100 per cent and 70 per cent respectively. The laboratory participated in two external quality assessment schemes for cholesterol testing during the course of the audit and all the results of these fell within the acceptable limits. The audit demonstrated that the Reflotron was too imprecise to be used to give accurate measurements of blood cholesterol. However, providing a suitable Reflotron result above which patients were sent for confirmatory laboratory testing was selected, it was an acceptable screening device in the detection of hypercholesterolaemia. Other Reflotron users should consider carrying out similar audits. PMID- 8422443 TI - Developments in the darkroom: a cross-sectional study of sickness absence, work related symptoms and environmental monitoring of darkroom technicians in a hospital in Glasgow. AB - Nine female technicians using both manual and automatic processing methods were matched by age, sex and smoking habits with controls from the occupational therapy department. All were interviewed using a questionnaire about symptoms experienced during the day in question and the preceding month. Sickness absence information for 1987 was obtained, and the working environment was assessed by standard occupational hygiene methods. The technicians and controls had similar numbers of episodes of sickness absence (23 versus 24, respectively) but aggregated substantially more days absence attributed to sickness (204 versus 41). The total number of 'monthly' symptoms experienced by the technicians was greater than the controls (44 versus 30) as was the number of 'daily' complaints (26 versus 6). In all cases, respiratory symptoms were responsible for about one third of problems, but statistical analysis (Wilcoxon matched pairs signed ranks tests) showed that the differences were not statistically significant. Occupational hygiene results were all within normal limits. PMID- 8422444 TI - Irritant contact dermatitis in warehouse employees. AB - A detailed survey of skin complaints amongst 114 airline employees working in a new warehouse revealed 26 cases of skin problems which originated during the 2 1/2 year operation. A clinical survey of broadly the same population confirmed 14 cases from 98 employees as chronic irritant contact dermatitis of the hands. The work involved the reception, unpackaging, inspection, repackaging and dispatch of aircraft parts. The source of the skin irritation was not to be found in the work itself. Rather, the presence among the employees of two severe cases of non occupational eczema, combined with the idea that incoming aircraft parts from foreign countries might be 'dirty' in some way, had caused a heightened perception of a risk of skin disease, and the frequency of hand washing had increased as a result. Over-frequent hand washing in a few employees had resulted in precisely what the warehouse staff had been trying to avoid. PMID- 8422445 TI - Tinnitus in noise-exposed workers. AB - Tinnitus is said to be a common complaint of workers who are exposed to noise. We studied the prevalence and characteristics of tinnitus in 647 noise-exposed workers who had been notified as cases of noise-induced deafness. One hundred and fifty-one had tinnitus, giving a prevalence of 23.3 per cent. The tinnitus was bilateral in 42.4 per cent of cases, and of high frequency in 44.4 per cent. In 23.8 per cent it was associated with other symptoms. About 30 per cent of those with tinnitus complained that it interfered with daily activities like telephone conversation and sleep. The workers with tinnitus had consistently higher hearing thresholds at both high and low frequencies than those with no tinnitus. This finding remained even after adjusting for differences in sex, age and ethnic group composition and in the noise exposure duration. Workers are often told that noise exposure causes deafness, but little is mentioned about tinnitus. Awareness of the possible occurrence of tinnitus may encourage workers to cooperate more actively in a company hearing conservation programme. PMID- 8422446 TI - Attacks on postmen in Northern Ireland. What features of the attacks are associated with prolonged absence from work? AB - The sickness absence records of 228 postmen who had been attacked on duty in Northern Ireland between 1985 and 1989 were examined. The pattern of absences due to psychological causes in the six months after the attack was recorded. The presence of relevant sickness absence and the amount was correlated to three specific features of the attacks, ie use of arms, use of violence and abduction. Armed attacks produced a significantly more frequent and larger amount of sickness absence. The proportion of victims taking any sickness absence after violent attacks was not significantly greater but those who did so were off work for much longer. Abduction of victims from the scene produced much more frequent absence from work but there was no significant difference in the duration of the absence. The 20 employees who were attacked for a second time during the study period showed a five-fold increase in the length of sickness absence taken. PMID- 8422447 TI - The health care industry. AB - This series of articles is concerned with technological change in industry over the last 20 years, its effect on the health of the worker, and the measures taken by the employer to control any risk arising therefrom. Technological change cannot be viewed in isolation. Although advances in technology and the introduction of new processes are the outward and most visible sign of change in work practices, in most cases it is managerial restructuring, political pressures, and changes in legislation which bring about far greater change and which in the long term may have a greater effect on the health of the worker. No industry illustrates this better than the National Health Service and more recently the rapidly expanding private health care sector. PMID- 8422448 TI - Approach to risk reduction in manufacturing firms in Australia. AB - A survey of the 100 largest manufacturing firms in Australia has revealed that there is a lack of emphasis on controlling the incidence of high severity injuries and diseases. In a considerable number of organizations, there appears to be a reliance on the attitude of employees and safety training to control risk. There seems to be a lack of knowledge among the respondents of information sources which can be used to assess both risk and proposed control measures. The results suggest that Robens type legislation may be of limited utility in controlling risk in the workplace and a more definitive approach to identifying hazards and instituting appropriate control measures should be provided to organizations. PMID- 8422449 TI - Tempora mutantur, et nos mutamur in illis. PMID- 8422450 TI - Should atopic employees be excluded from specific occupations? PMID- 8422451 TI - Does RSI exist? PMID- 8422452 TI - Where is the workplace and who is the practitioner? PMID- 8422453 TI - One view of the Japanese work ethic. PMID- 8422454 TI - Preparative regimens for autologous bone marrow transplantation. PMID- 8422455 TI - Hematopoietic growth factors and their receptors in acute leukemia. PMID- 8422456 TI - Topographical dissociation of BCL-2 messenger RNA and protein expression in human lymphoid tissues. AB - Immunohistochemistry and in situ hybridization with a synthetic oligonucleotide probe were used to compare the topographical distribution of BCL-2 proto oncogenic protein with that of its messenger RNA (mRNA) in normal lymphoid tissues, follicular lymphomas, and lymphoma-derived cell lines. In normal lymph nodes, BCL-2 protein was most abundant in the small lymphocytes of primary lymphoid follicles and the mantle zones of secondary follicles, virtually absent within germinal centers, and of variable abundance in many interfollicular cells. In contrast, the distribution of BCL-2 mRNA was roughly reciprocal to that of the protein with intense hybridization signal in germinal centers and almost none in mantle zones. Discordant BCL-2 RNA and protein levels were also observed in tonsillar epithelial cells and cortical thymocytes. Concordant and abundant expression of BCL-2 mRNA and protein was detected in biopsy tissues and cell lines from t(14;18)-carrying lymphomas. The contrasting distributions of BCL-2 protein and RNA in normal lymphoid tissues suggest that translational and posttranslational control mechanisms play a significant role in regulating BCL-2 protein levels in germinal center cells, epithelial cells, and cortical thymocytes. Concordant BCL-2 mRNA and protein levels in follicular lymphomas suggest that translational control mechanisms may be disrupted as part of the sequence of genetic changes that transforms normal lymphoid cells into neoplastic follicular lymphoma cells. PMID- 8422457 TI - Autologous bone marrow transplantation for acute myeloid leukemia using busulfan plus etoposide as a preparative regimen. AB - We have studied the use of a new preparative regimen for the treatment of patients in remission of acute myeloid leukemia (AML) with autologous bone marrow transplantation. Chemotherapy consisted of busulfan 1 mg/kg every 6 hours for 4 days (total dose, 16 mg/kg) on days -7 through -4 followed by an intravenous infusion over 6 to 10 hours of etoposide 60 mg/kg on day -3. Autologous bone marrow, treated in vitro with 100 micrograms/mL of 4-hydroperoxycyclophosphamide, was infused on day 0. We have treated 58 patients up to the age of 60 years, 32 in first remission, 21 in second or third remission, and 5 with primary refractory AML unresponsive to high-dose Ara-C, but achieving remission with aggressive salvage regimens. Of the first remission patients, there has been 1 treatment related death and 5 relapses. With median follow-up of 22 months, the actuarial relapse rate is 22% +/- 9% and disease-free survival is 76% +/- 9% at 3 years. Patients with favorable French-American-British (FAB) subtypes (M3 or M4 EO) did especially well, with no relapses seen in 15 patients observed for a median of 30 months. Actuarial relapse rate at 3 years was 48% for first remission patients with less favorable FAB subtypes. Of patients in second or third remission, there were 5 treatment related deaths and 4 relapses. With median follow-up of 22 months, the actuarial relapse rate is 25% +/- 11% and disease-free survival is 56% +/- 11% at 3 years. Four of five primary refractory patients died during treatment and 1 remains in remission with short follow-up. These preliminary data are very encouraging and, if confirmed, support the use of autologous purged bone marrow transplantation using aggressive preparative regimens as one approach to improve the outcome of adults with AML. PMID- 8422458 TI - Busulfan/etoposide--initial experience with a new preparatory regimen for autologous bone marrow transplantation in patients with acute nonlymphoblastic leukemia. AB - Current intensive chemotherapy for acute nonlymphoblastic leukemia (ANLL) results in a complete remission in the majority of patients. Unfortunately, the duration of remission is short and most of the patients will experience a relapse of their underlying disease. Autologous bone marrow (BM) transplantation is being explored as a treatment modality designed to improve relapse-free survival. We have conducted a phase II trial exploring the combination of busulfan (16 mg/kg) and etoposide (60 mg/kg) in an attempt to improve antitumor efficacy using this novel preparative regimen. To date, 50 patients (48 with ANLL and 2 patients with biphenotypic acute leukemia) have been treated. The first 20 patients received unmanipulated BM; 28 patients subsequently received 4-hydroperoxycyclophosphamide (4-HC) (60 micrograms/mL)-purged bone marrow, and 2 patients with biphenotypic acute leukemia received both 4-HC (60 micrograms/mL) and etoposide (5 micrograms/mL)-purged BM. Thirty-four patients were in first complete remission (CR1), 12 patients in second complete remission (CR2), and 4 patients in relapse. The median time from first complete remission to BM harvest was 3 months (range, 0.8 to 4) compared with median time of 2 months (range, 1.5 to 5.0) for patients in second complete remission. The median time from harvest to transplant was 1 month for both groups (range, 0.4 to 36). A median of 0.7 x 10(8) (range, 0.2 to 1.4) mononuclear cells were infused. Patients achieved an absolute neutrophil count of > or = 500/microL at a median of 26 days (range, 13 to 96), an untransfused platelet count > or = 20,000/microL at a median of 56 days (range, 15 to 278) and a sustained hematocrit > or = 30% at a median of 50 days (range, 19 to 116). Twenty-six patients are alive and in continued CR. Follow-up of the surviving patients ranged from 6 months to 66 months with a median follow-up of 31 months. Patients receiving purged BM have an actuarial disease-free survival of 57% with a relapse rate of 28% compared with patients receiving unpurged BM whose actuarial disease-free survival is 32% with a relapse rate of 62% (P = .06 for relapse rate). The most significant extramedullary toxicities for this regimen are hepatic and cutaneous (including mucositis). The BU/VP-16 regimen is associated with a significant proportion of patients surviving disease free, especially in the group receiving purged BM. Whether this regimen offers a substantial improvement in disease-free survival over currently used regimens will require a prospective randomized study. PMID- 8422459 TI - Interleukin-4 inhibits the lipopolysaccharide-induced expression of c-jun and c fos messenger RNA and activator protein-1 binding activity in human monocytes. AB - We studied the effect of interleukin-4 (IL-4) on the lipopolysaccharide (LPS) induction of two immediate early genes c-fos and c-jun. These genes encode proteins that form the dimeric complex activator protein-1 (AP-1), which is active as a transcriptional factor. Maximal accumulation of either c-fos and c jun messenger RNA (mRNA) occurred 30 minutes after LPS addition. When cells were treated with IL-4 for 5 hours before LPS activation, both the c-fos and the c-jun mRNA expression was decreased. The inhibition of c-fos and c-jun expression by IL 4 in LPS-treated cells was shown to be due to a lower transcription rate of the c fos and c-jun genes. IL-4 did not affect the stability of the c-fos and c-jun transcripts. Finally, using electrophoretic mobility shift assays, evidence was obtained that IL-4 inhibits LPS-induced expression of AP-1 protein. These data indicate that IL-4 suppresses the induction of transcription factors in human activated monocytes. PMID- 8422460 TI - Protein S enhances C4b binding protein interaction with neutrophils. AB - Protein S, an inhibitor of coagulation, interacts reversibly with C4b binding protein (C4bBP). The physiologic role of this complex formation remains unknown. In this study we examined the possibility that protein S would facilitate C4bBP binding to the surface of neutrophils where, in turn, this complex could help protect the cell from complement-mediated damage at the site of inflammation. Neutrophils bind approximately 60,000 protein S molecules per cell (kd = 140 nmol/L) in a Ca(2+)-dependent fashion. C4bBP also binds to neutrophils (23,000 sites per cell at physiologic C4bBP concentration), but this binding is not Ca2+ dependent. Protein S approximately doubled C4bBP binding, but protein S only influenced C4bBP binding in the presence of Ca2+. Protein S binding to neutrophils decreased approximately twofold in the presence of saturating concentrations of C4bBP. Neutrophil activation had only minor effects on the affinity and number of sites for protein S or the protein S-C4bBP complex. These results indicate that the protein S-C4bBP complex binds to the neutrophil surface where it can presumably modulate complement assembly on the cell surface. PMID- 8422461 TI - Determination of adhesion force between single cell pairs generated by activated GpIIb-IIIa receptors. AB - A biophysical approach was used to directly determine the avidity of the junction between two Chinese hamster ovary (CHO) cells bearing recombinant GpIIb-IIIa in the presence and absence of fibrinogen. Micromanipulation was used to induce conjugation of the cell pairs with or without activating the GpIIb-IIIa molecules with monoclonal antibody (MoAb) 62. Activation of GpIIb-IIIa caused an increase in the force required to separate the conjugates. The molecular bonding force between cells bearing activated GpIIb-IIIa and fibrinogen molecules was found to be 2.1 x 10(-7) dyne, which is 3.7 times higher than that between nonactivated GpIIb-IIIa and fibrinogen (5.7 x 10(-8) dyne). The results provide a quantitative assessment of the molecular bonding force between fibrinogen and the GpIIb-IIIa expressed on cell surface. The findings indicate that the activation of GpIIb IIIa leads to an increase in the adhesive force in CHO cell aggregation by increasing the strength of the GpIIb-IIIa-fibrinogen bonds rather than the number of these bonds. PMID- 8422462 TI - High serum and ascitic soluble interleukin-2 receptor alpha levels in advanced epithelial ovarian cancer. AB - This study was undertaken to determine if advanced epithelial ovarian cancer was associated with increased serum and ascitic levels of soluble interleukin-2 receptor alpha (sIL-2R alpha). Serum and ascitic fluid samples from 23 ovarian cancer patients were analyzed for sIL-2R alpha using an enzyme-linked immunosorbent assay and compared with the serum and peritoneal levels in 18 normal females. The samples were analyzed for CA-125 levels using a radioimmunoassay and the total protein was also measured. Normal individuals had low serum levels of sIL-2R alpha (367.5 +/- 44.6 U/mL), with similar levels of sIL-2R alpha in the normal peritoneal fluid (438.6 +/- 48.8 U/mL). In contrast, the serum and ascitic fluid levels in ovarian cancer patients were significantly higher (746.7 +/- 82.9 U/mL, P = .0006; 2,656.7 +/- 373.7 U/mL, P = .00002, respectively). The results for sIL-2R alpha were also significant when the levels were expressed per milligram of total protein. More importantly, in almost every ovarian cancer patient the ascitic sIL-2R alpha level far exceeded the serum level, a pattern also observed for CA-125. There was no correlation between the serum and ascitic sIL-2R alpha levels, or between the serum and ascitic CA-125 levels. Although the serum levels of sIL-2R alpha and CA-125 were elevated in the same patient, overall there was no correlation between the serum sIL-2R alpha and serum CA-125 levels, either when the levels were expressed in absolute units or per milligram of total protein. Similarly, there was no correlation between sIL 2R alpha and CA-125 levels in individual ascitic samples. While CA-125 levels may reflect an independent index of tumor burden, these results suggest that selective accumulation of sIL-2R alpha in the ascites may be one of the factors associated with the known nonresponsiveness of the infiltrating lymphocytes against ovarian carcinoma cells. PMID- 8422463 TI - Human immunodeficiency virus-1 infection of the human promyelocytic cell line HL 60: high frequency of low-level infection and effect of subsequent cell differentiation. AB - As a model system to study the infection of early myeloid cells by human immunodeficiency virus-1 (HIV-1), we have infected the human promyelocytic cell line, HL-60, with a recombinant selectable HIV-1 clone. A fully infected population showed a relatively high frequency of low-level infection, with 40% of subcloned cells being negative by reverse transcriptase and p24 indirect immunofluorescence analysis and displaying only low levels of supernatant p24. The same treatment of a T-lymphoid cell line produced 100% productive infections. HIV-1 infection of HL-60 did not appear to alter the state of differentiation of the cells, as assessed by surface antigen expression, regardless of the level of viral expression. Furthermore, infected cells were able to respond normally to chemical inducers of differentiation. Induction of differentiation towards monocyte/macrophages by phorbol myristate acetate activated the HIV-1 long terminal repeat in a transient transfection system, and there was a corresponding increase in viral production from the infected subclones. Granulocytic differentiation, as stimulated by dimethyl sulfoxide or retinoic acid, had no effect on long terminal repeat activity and did not stimulate viral replication. These data suggest that low-level HIV-1 infections may be established at a relatively high frequency in myeloid precursor cells, and that different pathways of promyelocytic differentiation vary in their ability to stimulate HIV-1 replication. PMID- 8422464 TI - Subtractive cDNA cloning of a novel member of the Ig gene superfamily expressed at high levels in activated B lymphocytes. AB - Using subtractive cDNA cloning we have isolated a series of cDNA clones that are exclusively or selectively expressed in B lymphocytes. mRNA transcripts from one such cDNA clone, referred to as BL11, were found to be expressed at low levels in RNA from normal B lymphocytes, but at very high levels in RNA from in vitro activated B lymphocytes. One major 2.5-kb BL11 mRNA transcript was detected, while low levels of 4.8-, 1.8-, and 1.6-kb transcripts were also found. BL11 mRNA transcripts were absent or present at low levels in RNA prepared from resting or mitogen activated T cells, a variety of lymphoid cell lines including several B cell lines, and several different tissues. Low levels of BL11 transcripts were found in poly(A) RNA purified from brain and lung. A study of the kinetics of BL11 mRNA accumulation in B lymphocytes stimulated in vitro with Staphylococcus aureus Cowan strain I showed a rapid induction of BL11 mRNA within 2 hours of stimulation with peak expression by 16 hours and a mild decrease with time following the peak levels. Consistent with the in vitro data, in situ hybridization using antisense BL11 RNA probes and human tonsillar tissue localized BL11 transcripts in B-cell-enriched areas. Multiple BL11 cDNA and genomic clones were isolated and sequenced to complete and verify the BL11 cDNA sequence (2,404 bp). A 615-nucleotide open reading frame predicted to encode for a 205-amino acid protein with a molecular weight of 23 Kd was identified. Search of protein data bases with the predicted BL11 protein showed homologies to several members of the Ig superfamily. Analysis of the predicted protein showed a likely signal peptide, a single membrane spanning region, and one V-like Ig domain with three predicted n-glycosylation sites. Southern blot analysis of human genomic DNA suggested that BL11 is a single copy gene without evidence of rearrangement. Primer extension and S1 nuclease mapping identified four tightly clustered transcriptional start sites approximately 40 bp upstream of the predicted translation start site. The first 270 bp of the promoter region were sequenced and found to contain a CATAA box rather than a TATAA box and several DNA motifs found in activation genes. BL11 should prove to be an interesting gene that likely encodes for a protein involved in B-cell activation. PMID- 8422465 TI - Regulation of myeloblastin messenger RNA expression in myeloid leukemia cells treated with all-trans retinoic acid. AB - Retinoic acid is known to induce differentiation of human myeloid leukemia cells in vitro. Recently, all-trans retinoic acid has been used to induce remissions in patients with acute promyelocytic leukemia, probably through differentiation of the leukemia cells. Myeloblastin (mbn) is a protease that has been identified in the human leukemia cell line HL-60. Downregulation of this protease can inhibit proliferation and induce differentiation of HL-60-derived leukemia cells. Here we have investigated the regulation of mbn messenger RNA (mRNA) expression in two human leukemia cell lines, HL-60 and NB4, treated with all-trans retinoic acid. Under this treatment, downregulation of mbn mRNA was observed in both cell lines, but was considerably delayed in NB4 cells that carry the t(15;17) translocation characteristic of acute promyelocytic leukemia. We have found that multiple mechanisms were involved in the control of mbn mRNA expression. These mechanisms were different in HL-60 and NB4 cells. Our results show that in HL-60 cells, all trans retinoic acid rapidly decreased transcription of mbn. In contrast, in the t(15;17)-positive NB4 cells treated with all-trans retinoic acid, upregulation of mbn mRNA expression was followed by a late downregulation, both achieved via posttranscriptional mechanisms. PMID- 8422466 TI - Retrovirally transduced antisense sequences stably suppress P210BCR-ABL expression and inhibit the proliferation of BCR/ABL-containing cell lines. AB - There is now strong evidence that the BCR-ABL gene product of the Philadelphia chromosome (P210) plays a crucial role in the pathogenesis of chronic myeloid leukemia (CML). We have previously shown that introduction of antisense oligonucleotides into K562 cells could transiently block the expression of P210 and specifically inhibit cellular growth in culture. In this report, we describe the use of a retroviral vector to introduce selected antisense and sense sequences, first into murine B10 cells, previously rendered interleukin-3 (IL-3) independent by transfection of BCR-ABL sequences, and second into K562 cells. The antisense transcripts generated under the control of MoMLV promoter specifically killed B10 cells in the absence of IL-3 and inhibited P210 expression almost completely. In K562 cells, the antisense sequences led to a dramatic reduction of P210 expression and increased their doubling time by more than twofold. This effect was not reversed by the addition of exogenous IL-3 to the culture medium. Control HeLa or HL60 cells infected with the same constructs did not show any change in proliferation rate, despite abrogation of the normal BCR gene products. Rather unexpectedly, P210 suppression was not lethal in K562 cells, showing that such a cell line does not rely entirely on the expression of P210 for surviving, but depends on it as far as growth properties are concerned. We conclude that this approach can successfully achieve stable suppression of the oncogenic protein P210 and may be used to study further the mechanisms by which P210 is transforming cells. The effect on fresh CML cells in bone marrow cultures remains to be assessed before we can tell whether this technique may be used for selective suppression of leukemic hematopoiesis in vitro. PMID- 8422467 TI - Function of integrin in duodenal mucosal uptake of iron. AB - A mechanism for the absorption of inorganic iron in the small intestine is described in which integrins appear to play an important role in the passage of iron across microvillous membranes. Biochemical isolates from microvillous preparations of duodenum from rats dosed with radioiron showed radioactivity concentrated in integrins. The presence of integrins on mucosal surfaces of duodenal cells was confirmed by immunofluorescent microscopy using anti-integrin monoclonal antibodies. Immunoprecipitation methods were used to show that microvillous radioiron was precipitated with anti-integrin antibodies and that mobilferrin, a 56-Kd cytosol iron-binding protein, coprecipitated with integrins. We postulate from these data that the mucosal uptake of iron from the gut lumen is mediated via an integrin-mobilferrin pathway. PMID- 8422468 TI - Dependence of the permanent deformation of red blood cell membranes on spectrin dimer-tetramer equilibrium: implication for permanent membrane deformation of irreversibly sickled cells. AB - Red blood cells (RBCs) in sickle cell anemia, transformed into a sickled shape by prolonged deoxygenation, or normal RBCs deformed by a prolonged micropipette aspiration become permanently stabilized in their abnormal shape. This semisolid plastic behavior is thought to involve an irreversible reorganization of the membrane skeleton, but the exact nature of this skeletal rearrangement is not known. In this study, we first asked whether the irreversible deformation is associated with a permanent stretching of the skeletal network, and then whether it is due to a rearrangement of skeletal components involving a disruption of pre existing protein associations and the subsequent reassociation of new protein contacts. Having found no ultrastructural evidence of stretching of the skeletal lattice in membranes derived from permanently deformed RBCs, we addressed the possibility of reorganization of the proteins of the membrane skeleton. We examined the temperature dependence of irreversible cell deformation to see if it correlated with the known temperature dependence of spectrin tetramers to dimer dissociation and reassociation. Testing the shape irreversibility of both deoxygenated reversibly sickled cells and Nucleopore-aspirated normal cells, we found that both types of cells became permanently deformed when the prolonged incubation of applied force or deoxygenation was performed at 37 degrees C, the temperature at which spectrin tetramers were free to dissociate and reassociate. In contrast, both types of cells were able to regain their original discocytic shape if the prolonged incubation was performed at the lower temperature: at less than 13 degrees C instead of 37 degrees C. Furthermore, normal RBCs were incubated with inosine and pyruvate to elevate intracellular 2,3 diphosphoglycerate, the polyanion shown to destabilize spectrin-actin-protein 4.1 association. This did not result in a promotion of irreversible deformation of these cells. We conclude that the irreversible cell deformation observed at physiologic temperature is associated with a skeletal rearrangement through dissociation of spectrin tetramers to dimers and a subsequent reassociation of dimers to tetramers in the new (deformed) configuration. These findings may explain a permanent stabilization of irreversibly sickled cells in their abnormal shape in vivo. PMID- 8422469 TI - Fetal hemoglobin induction with butyric acid: efficacy and toxicity. AB - Butyric acid induces fetal hemoglobin (HbF), a property of potential therapeutic advantage in patients with disorders of globin chain synthesis. We performed dose escalation studies of this compound in baboons to assess whether clinically significant increases in HbF are achievable, and to define the associated toxicities. Additionally, the effect of butyrate in combination with erythropoietin on HbF induction was assessed. HbF induction in response to butyrate was dependent on the dose and duration of treatment. Doses of butyrate less than 4 g/kg/d were associated with minimal toxicity (hypokalemia) and significant HbF induction in these nonanemic animals, with 1 g/kg/d producing an increase in HbF-containing reticulocytes (F reticulocytes) from 0.9% to 8.7% and an increase in HbF from 0.8% to 1.4%. A dose of 2 g/kg/d resulted in an increase in F reticulocytes from 2.1% to 27.8% and an increase in HbF from 0.7% to 2.2%. Doses of 4 g/kg/d in another animal produced an increase in F reticulocytes from 1% to 21.6% and in HbF from 1.9% to 5.3%. Infusions in excess of 4 g/kg/d were complicated (after a variable amount of time) by a decreased level of alertness (caused by hyperosmolality or butyrate itself) and hematologic toxicity (with declines in reticulocyte, white blood cell, and platelet counts). Prolonged infusions of high doses of butyrate (8 to 10 g/kg/d) were associated with peak F reticulocyte percentages reaching 38% to 64.5% and HbF reaching levels in excess of 20%. These high doses (8 to 10 g/kg/d) were complicated in two animals with a striking and unique neuropathologic picture and, in one animal, multiorgan system failure. Erythropoietin in combination with butyrate, induced F reticulocytosis in an additive manner. We conclude that butyric acid is a strong inducer of HbF, particularly when administered in combination with erythropoietin. As chronic toxicities remain undefined, patients in future clinical trials of this and similar compounds should be monitored closely for evidence of neurologic toxicity. PMID- 8422470 TI - Downregulation of the anti-HLA alloimmune response by variable region-reactive (anti-idiotypic) antibodies in leukemic patients transfused with platelet concentrates. AB - Approximately 30% to 40% of patients with acute leukemia receiving repeated pooled random-donor platelet transfusions develop anti-HLA alloantibodies. Over time, however, serum anti-HLA concentrations decrease in approximately 50% of these patients, despite continued exposure to platelet and/or red blood cell transfusions. Using an enzyme-linked immunosorbent assay to measure serum Igs, the present study demonstrates that the sera of 67% of 82 transfused patients exhibiting a decrease in anti-HLA contain antibodies (anti-idiotypes) that react with the variable (V) region of anti-HLA antibodies. Anti-HLA binding to platelet membranes could be inhibited by these serum antibodies in 36% of the patients, indicating they had paratope-related reactivity. Protein G sepharose absorption showed that the anti-HLA V region-reactive antibodies were IgG. Of the 43 patients who had a decrease in anti-HLA levels, that were 16 whose anti-HLA decreased to undetectable levels; 7 (44%) developed anti-idiotypic antibodies that could specifically inhibit their own previously anti-HLA-positive serum. In contrast, antibodies with reactivity to the V region of anti-HLA antibodies (anti idiotypes) were not demonstrable in patients who developed anti-HLA that did not decrease or disappear. The findings suggest that the development of anti-HLA V region-reactive antibodies (anti-idiotypic antibodies) correlates with a decrease in anti-HLA antibody formation in patients multiply transfused with platelet concentrates. The observations indicate that anti-idiotypic antibodies may downregulate alloimmune responses in patients undergoing repeated allostimulation during platelet transfusion therapy. PMID- 8422471 TI - Unrelated donor marrow transplantation therapy for chronic myelogenous leukemia: initial experience of the National Marrow Donor Program. AB - In the interval from December 1987 to November 1990, 196 consecutive patients with chronic myelogenous leukemia (CML) received unrelated donor marrow transplantation using marrow procured by the National Marrow Donor Program (NMDP) at 21 NMDP-affiliated marrow transplant centers. Baseline donor and recipient data as well as follow-up data were obtained systematically in all cases by the NMDP. The median interval from the initiation of a search for an unrelated donor to bone marrow transplantation was 8.4 months (range, 1.7 to 34.6 months). Median age of the recipients was 33.3 years (4.5 to 54.5 years). Seventy-five recipients were female and 121 were male. At time of transplant, 115 patients were in chronic phase, 51 in accelerated phase, 14 in blast crisis, and 16 in a second or subsequent chronic phase. In 133 cases, donors and recipients were identical at the HLA A, B, and DR loci using standard serologic typing, and in 63 cases, there was nonidentity at one HLA locus. Patients were prepared for transplantation with a combination of high-dose chemotherapy and total body irradiation (N = 169) or with high-dose chemotherapy only (N = 27). Thirty-five patients received marrow depleted ex vivo of T lymphocytes, whereas 161 patients received non-T-depleted marrow. One hundred seventy-four of 196 patients engrafted (absolute neutrophil count > or = 500/mm3 for 3 consecutive days). The median time to engraftment was 22 days (6 to 69 days). Twenty-two patients failed to engraft, and an additional 10 patients experienced late graft failure. The incidence of grades III or IV acute graft-versus-host disease (GVHD) was 0.54 +/- 0.10, and that of extensive chronic GVHD was 0.52 +/- 0.12. A lower incidence of both grades III and IV acute GVHD (P = .0003) and of extensive chronic GVHD (P = .01) were independently associated with use of T-depleted marrow. The actuarial incidence of hematologic relapse at 2 years is 0.11 +/- 0.06. The 2-year actuarial incidence of disease free survival for patients transplanted in first chronic phase within 1 year of diagnosis is 0.45 +/- 0.21, in chronic phase more than 1 year from diagnosis is 0.36 +/- 0.11, in accelerated phase is 0.27 +/- 0.12, in second or subsequent chronic phase is 0.22 +/- 0.21, and in blast crisis is 0. Fifteen of 55 patients transplanted at 40 to 50 years of age survive.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8422472 TI - Detection of residual leukemia by the polymerase chain reaction and sequence specific oligonucleotide probe hybridization after allogeneic bone marrow transplantation for AKR leukemia: a murine model for minimal residual disease. AB - Disease relapse after allogeneic bone marrow transplantation (BMT) is a major cause of treatment failure and is thought to evolve from clinically occult residual disease in the recipient. However, the demonstration of minimal residual disease (MRD) in individual patients is of uncertain prognostic significance because the detection of residual disease has not consistently correlated with subsequent relapse. Moreover, the optimal therapeutic approach in patients with MRD after allogeneic BMT is unknown. The study of these issues has been hindered by the lack of clinically relevant animal models. In this report, we characterize a novel murine model for the study of MRD after allogeneic BMT. This model was designed to simulate high-risk BMT in humans in which patients receive transplants in relapse and disease recurrence in the major cause of treatment failure. The H-2-compatible, mixed lymphocyte culture nonreactive murine strains, AKR (H-2k) and CBA (H-2k), were chosen to parallel marrow transplants from HLA matched siblings, which represent the majority of allo-transplants in humans. Male AKR leukemia cells were used in female donor/host chimeras permitting the Y chromosome to serve as a leukemia-specific marker for MRD. Detection of residual male leukemia cells in the peripheral blood of the primary host was facilitated by use of the polymerase chain reaction (PCR) and sequence-specific oligonucleotide probe hybridization (SSOPH). Use of PCR/SSOPH was highly predictive of clinical outcome (relapse or cure) in animals receiving transplants (P < .00002) and detected disease recurrence earlier than comparative flow cytometric analysis studies. This murine model will be useful in evaluating the efficacy of therapeutic strategies aimed at reducing disease relapse posttransplant and can be adapted to other transplant murine tumor systems for the study of MRD. PMID- 8422473 TI - Complete remission after fludarabine for chronic lymphocytic leukemia. PMID- 8422474 TI - CD45 expression and prognosis in acute lymphoblastic leukemia. PMID- 8422475 TI - Bence Jones protein in primary systemic amyloidosis. PMID- 8422476 TI - Rapid diagnosis of hemoglobin constant spring and hemoglobin E by amplified created restriction sites. PMID- 8422478 TI - Early ABRs in infants undergoing assisted ventilation. AB - ABR was performed on 42 preterm infants undergoing assisted ventilation with conventional or high-frequency oscillatory ventilation (HFOV). ABRs from these very young neonates were evaluated to further detail the emerging response and to determine if type of ventilation or other perinatal factors had effects on the ABR. While responses were present down to 26 weeks gestational age, the only factors which appeared related to absent ABRs were birthweight and gestational age. PMID- 8422477 TI - Omission rates of young and elderly listeners in word recognition testing. AB - Elderly listeners, as a group, do not perform as well as younger listeners on tests of word recognition, particularly in the presence of competing noise. It has been suggested that elderly listeners may be less willing to guess at items when they are unsure of the correct response, and as a result omit more responses. Omitting responses on word recognition tests may deflate word recognition scores. To test whether elderly listeners actually do omit more responses on word recognition tests, a group of elderly normal-hearing adults was compared to a group of young normal-hearing adults. To promote guessing, ipsilateral multitalker babble was presented at individually determined levels that approximated 50 percent performance. The subjects responded to two lists of NU6 words, and numbers of omissions were recorded. The elderly group did not omit significantly more responses to word recognition test items than the younger group. Variability was high within both groups. PMID- 8422479 TI - Test-retest reliability of a shortened version of the hearing aid performance inventory. AB - Sixty-four elderly hearing aid users completed a shortened version of the Hearing Aid Performance Inventory (HAPI) on two occasions, separated by 1 to 2 months. The test-retest correlation was .80 for the overall scale and ranged from .70 to .85 for the four factor ratings. The mean overall rating and the mean ratings on each of the four factors demonstrated nonsignificant shifts from test to retest. The 95 percent confidence intervals for the overall scale and for each factor are presented for interpretation of differences in HAPI ratings between aided conditions. PMID- 8422480 TI - Reliability of threshold estimation in hearing-impaired adults using the AMFR. AB - The amplitude modulation-following response (AMFR), an auditory evoked potential elicited by continuous amplitude-modulated tones, can be recorded for carrier frequencies across the audiometric range. AMFR thresholds (based on the amplitude spectra of the responses) have been found to closely follow behavioral thresholds in six normal-hearing and four hearing-impaired adults. In the current work, we studied the reliability of the AMFR as an index of behavioral low-frequency (500 and 1000 Hz) thresholds in a larger sample (n = 16) of hearing-impaired adults with mild to moderate hearing loss. The AMFR amplitudes and detection thresholds were strongly correlated across the two recording sessions, and AMFR thresholds obtained in both sessions were strongly correlated to behavioral thresholds. The average absolute difference between the AMFR and behavioral thresholds was 7.3 dB and 6.4 dB for 500 Hz and 1000 Hz, respectively. PMID- 8422481 TI - Ability to achieve gain/frequency response and SSPL-90 under three prescription formulas with in-the-ear hearing aids. AB - Custom in-the-ear (ITE) hearing aids (standard linear amplifiers with single-pole filter low-frequency tone control and a class A amplifier output stage) were fit to 90 ears using the revised National Acoustics Laboratories' formula (NAL-R), and to 20 ears each using Prescription of Gain/Output II (POGO II) and Memphis State University (MSU) formulas. Both real-ear insertion gain and 2-cc coupler gain were evaluated. Examination of differences between prescribed gain and that actually achieved in the fittings revealed that too much gain was often given in the low- and mid-frequency range and insufficient gain in the high frequencies. There was little difference among the formulas in the degree of deviation from target. For some fittings, the deviation resulted in poorer predicted speech recognition scores (modified Speech Transmission Index). For the POGO II and MSU methods, underfitting of prescribed SSPL-90 values was far more common than overfitting. PMID- 8422482 TI - Gender affects audiometric shape in presbyacusis. AB - A review of large-scale surveys of hearing over the past 50 years reveals a "gender-reversal" phenomenon in the average audiograms of the elderly. Above 1 kHz males show greater average loss than females, but below 1 kHz females show greater average loss than males. The effect increases with both age and degree of hearing loss. The difference is present whether or not the elderly persons complain of a hearing problem and remains after persons with a history of noise exposure are excluded from the analysis. A possible explanation, based on the greater likelihood of cardiovascular disease in the elderly female, is considered. PMID- 8422483 TI - Real-ear measures in evaluation of frequency response and volume control characteristics of telephone amplifiers. AB - The spectral frequency response, frequency response range, and volume control linearity of five telephone amplifiers were examined using real-ear measures. All measurements were performed in KEMAR's (Knowles Electronics Manikin for Acoustic Research) ear canal using a composite speech-shaped waveform as the stimulus. Spectral frequency response and response range of each device was obtained at four volume control settings and compared to those of a standard telephone receiver. Only two of the amplifiers replicate the spectral frequency response of the standard receiver and show an increase in the amount of gain provided with increasing volume control rotation. The remaining three amplifiers show a more restricted spectral frequency response and response range when compared to those of the standard receiver. The volume control characteristics of the amplifiers were somewhat more uniform. Overall results indicate that the spectral frequency response and response range of telephone amplifiers can be objectively evaluated using real-ear measures, and these measures are essential in determining the usefulness of certain telephone amplifying devices. PMID- 8422484 TI - Speech-spectrum analysis of Mandarin: implications for hearing-aid fittings in a multi-ethnic society. AB - Using similar recording and analysis techniques, the long-term average speech spectra of English and Mandarin were compared in order to identify difference that might influence hearing-aid fitting strategies. Despite the well-documented pitch contour differences between English and Mandarin, no significant difference was found in the long-term average spectral analysis. Implications for hearing aid prescription are discussed. PMID- 8422485 TI - Patulous eustachian tube identification using tympanometry. AB - The patulous, or nonclosing, eustachian tube is believed to affect as many as 7 percent of all adults, causing physical and psychological difficulties. This paper provides a brief review of the literature regarding the patulous eustachian tube (PET), its symptoms, precipitating conditions, incidence, diagnosis, and current medical management. Several case studies are also presented to illustrate the use of tympanometry in PET identification. PMID- 8422486 TI - Multisensory verbal communication method. PMID- 8422487 TI - Cardiopulmonary bypass in neonates weighing less than 2.5 kg: analysis of the risk factors for early and late mortality. AB - A low weight has been implicated as one of the major reasons for deferring cardiac surgical procedures on cardiopulmonary bypass (CPB), resorting instead to palliative procedures. The purpose of this study is to analyze the risk factors associated with an increased mortality for surgery using CPB in small infants, and to compare this group with infants weighing over 2.5 kg. Between 1979 and 1990, 60 infants weighing less than 2.5 kg underwent surgery on CPB at our institute. The mean age was 17.9 days (range 1 to 68 days, SD = 16.4) and the mean weight was 2.1 kg (range 1.2 to 2.5, SD = 0.35). Eight had univentricular and 52 had biventricular repairs. The hospital mortality was 16.5% (70% CL 11.5% to 22.7%). The actuarial survival over 100 months was 67.5% (including hospital mortality). Over the same period, 946 patients under the age of 6 months but weighing more than 2.5 kg underwent surgery on CPB with a lower hospital mortality of 7.1% (70% CL 6.3% to 8.0%, p < 0.006). The risk factors associated with an increased risk of early death in the group weighing less than 2.5 kg were analyzed using logistic regression. They were: (1) presence of preoperative metabolic acidosis; (2) univentricular versus biventricular repair; and (3) duration of CPB. Within the less than 2.5 kg group, low weight per se did not affect the outcome adversely. The main cause of late death was the presence of associated medical conditions, notably bronchopulmonary dysplasia and tracheo bronchomalacia. PMID- 8422488 TI - Blood conservation and autotransfusion in cardiac surgery. AB - Blood conservation techniques of withdrawal of blood just before surgery, intraoperative blood salvage with Solcotrans (Solco Basle [UK] Ltd.), and profound hemodilution were used in 14 patients undergoing open heart surgery (group I) and compared with equally matched 14 patients who had profound hemodilution only during cardiopulmonary bypass (CPB) and acted as controls (group II). Group I patients required a mean of 255 mL of homologous blood per patient to achieve a target hemoglobin of 8 g/dL compared to group II patients who required a mean of 1,011 mL per patient (p = 0.0001). By using autologous blood there was a marked reduction in homologous blood exposure. Eight patients in group I and two patients in group II required no homologous blood. No adverse events occurred. In the process of conservation of blood in open heart surgery, we found the combination of the above techniques used in Group I patients to be safe and effective. PMID- 8422489 TI - The endothelium: a key to the future. AB - The vascular endothelium is a complex modulator of a variety of biological systems and may well be the key to definitive success in the treatment of cardiovascular disorders. Surgically-induced endothelial injury may occur preoperatively during cardiac catheterization and intraoperatively from mechanical manipulation, ischemia, hypothermia, and exposure to cardioplegic solutions. The normal endothelium is antithrombogenic and yet promotes platelet aggregation and coagulation if injured. Vasospasm, occlusive intimal hyperplasia, and accelerated arteriosclerosis can also all occur as a result of endothelial injury. Furthermore, endothelial injury is harmful even in the absence of disruption of its monolayer integrity. Thus, preservation of the endothelium should be an additional objective for all cardiovascular surgeons. Synthetic vascular grafts, cardiac valves, and artificial ventricles do not spontaneously endothelialize and thus usually require some form of anticoagulation to maintain patency. Hence, endothelialization of prosthetic implants became an attractive concept. A number of different methods of obtaining an endothelial lining of prosthetic material has since been developed; these include facilitated endothelial cell migration, and endothelial cell seeding by using either venous or microvascular endothelial cells. Manipulating the endothelium might well provide the next major advancement for therapeutic and preventive measures for cardiovascular disease. PMID- 8422490 TI - Initial report of the Working Group on Frontiers in Perioperative Myocardial Management. AB - An informal working group on perioperative myocardial management has been formed, which is predominantly composed of cardiac surgeons and anesthesiologists. The first meeting was introduced with a reminder that perioperative ischemia is still an important component of cardiac operations when sophisticated tools are used to monitor its presence and physiological effects. Moreover, perioperative ischemia is associated with adverse outcomes in many instances. The discussion touched upon generalized problems that included leukocyte activation, oxidant stress, warm cardioplegia, warm heart surgery, echocardiographic assessment of coronary blood flow, vascular endothelial responses to cardiac operations and cardiopulmonary bypass, myocardial stunning, substrate enhancement of cardioplegic solutions, tools for assessing perioperative ventricular performance, and detection of perioperative myocardial ischemia. It was agreed that a highly focused program on the agenda of myocardial stunning would be the topic for the next meeting of the group, with the results to be published in a future issue. PMID- 8422491 TI - The subscapular artery: an alternative conduit for coronary bypass. AB - The search for an alternative conduit for coronary bypass has prompted our use of the subscapular artery for coronary bypass via a left thoracotomy in three patients. Historically, this artery in conjunction with muscle flaps has been used successfully as a free graft. Its size and length have proved feasible for coronary bypass to the circumflex coronary artery system. All three patients have had excellent clinical results lasting from 6 to 21 months. PMID- 8422492 TI - Does coronary endarterectomy adversely affect the results of bypass surgery? AB - Coronary endarterectomy (TEA) is performed infrequently during coronary artery bypass graft (CABG) surgery due to the impression that it results in higher rates of myocardial infarction (MI), operative mortality (OM), and poor long-term outcome. To assess the effectiveness of TEA, 1,228 patients undergoing isolated CABG between 1982 and 1989 were evaluated. The incidence of OM (3.2%) and MI (6.0%) following TEA was similar to conventional CABG (OM = 3.8%, MI = 5.5%, p = NS). The incidence of low output syndrome (LOS, 15.1%) and intraaortic balloon pump insertion (IABP, 4.5%) following TEA was similar to conventional CABG (LOS = 12.6%, IABP = 6.0%, p = NS). The highest level of the cardiac specific isoenzyme (CK-MB) released following surgery was similar for the TEA group (46 +/- 49) and conventional CABG group (42 +/- 44, p = NS). Ventricular dysfunction, urgent surgery, left main stenosis, advanced age, and reoperative surgery were similar in the TEA and conventional CABG groups. At a mean follow-up of 4.2 years, 65.6% of all TEA patients were free of angina, 44.4% were gainfully employed, and 62% were in New York Heart Association Class I. The incidence of late myocardial infarction was 5.4%. The 5-year actuarial survival was 90%. Patients with double TEA and limited TEA ( < or = 3 cm TEA specimens) tended to have a lower 5-year survival. With strict criteria for selection of TEA patients and with significant technical experience, the short- and long-term results of TEA are comparable to conventional CABG. PMID- 8422493 TI - The pericardium reinforced suture annuloplasty: another tool available for mitral annulus repair? AB - From May 1985 to May 1992, 169 patients underwent surgery for mitral valve repair. In 87% of these patients, the valve reconstruction involved the mitral annulus. At the beginning, in an effort to preserve systolic movement of the annulus and avoid the implant of prosthetic materials inside the heart, we mainly used simple suture annuloplasty in 66 patients. When we reoperated upon three patients only a few months after reconstruction of the mitral valve for a dehiscence of the suture annuloplasty, we decided to perform ring annuloplasty with the Carpentier ring in 23 patients. Though we have not seen any problems with the Carpentier ring in our series, we performed a suture annuloplasty reinforced with a strip of autologous pericardium to eliminate all prosthetic material. Since introducing this technique in 1989, we have used this annuloplasty in 58 patients. We have not observed any dehiscence of the suture or other complications related to this procedure. Three patients with a pericardial annuloplasty underwent reoperation for other reasons; the autologous pericardium was perfectly attached to the annulus, covered by a smooth layer of fibrous tissue without calcification. After this encouraging initial experience, we believe that long-term follow-up is necessary to confirm that autologous pericardium is an effective method of mitral annulus repair. PMID- 8422494 TI - Five-year experience with the Medtronic Hall prosthesis in isolated aortic valve replacement. AB - We reviewed clinical data in 216 patients who underwent isolated aortic valve replacement with the Medtronic Hall prosthesis. Between January 1983 and December 1990, a total of 216 prosthetic valves were implanted in 180 males and 36 females. Preoperatively, 45.5% of patients were in New York Heart Association (NYHA) Class III to IV. Cumulative follow-up was 682 years, and 3.2% of patients were lost to follow-up. The actuarial 5-year survival rate was 90% for the whole group. All the patients were anticoagulated with aceno-coumarol (Sintrom). There were no cases of structural dysfunction and one patient presented with valve thrombosis. The Medtronic Hall valve has a low rate of thromboembolic events without structural failure. It is an excellent device for aortic valve replacement. PMID- 8422495 TI - Modern cardiac valve devices--bioprostheses and mechanical prostheses: state of the art. AB - The choice of bioprostheses and mechanical prostheses as valvular substitutes for cardiac valve replacement surgery has existed for over 20 years. The extensive developments over the past three decades have been introduced to reduce or eliminate valve related complications, namely thromboembolism, anticoagulant related hemorrhage, and structural failure, as well as to optimize hemodynamic performance. The biological valvular prostheses, namely porcine aortic or bovine pericardium, have been developed with tissue preservation, together with stent designs, that contribute to preservation of anatomical characteristics and biomechanical properties of the leaflets. The mechanical prostheses have been developed to eliminate structural failure, to facilitate prevention of blood status and thrombus formation, to facilitate radiopacity for evaluation of prosthesis function, and to facilitate intraoperative leaflet positioning. The implantation of the various present generation bioprostheses and mechanical prostheses requires special considerations to avoid technical complications and support ventricular performance. The studies of biological and mechanical prostheses, both randomized and nonrandomized, as well as specific prosthesis assessments, have contributed to the establishment of indications for types of prostheses. Bioprostheses have a high risk of structural failure and reoperation, while mechanical prostheses have a high risk of thromboembolism and anticoagulant hemorrhage. Within the bioprostheses population, the risk factors for structural valve deterioration are younger age and mitral prosthesis. Older patients (> 65 years of age) have a greater risk of valve related complications with mechanical prostheses, while younger patients (< 40 years of age) are at greater risk with bioprostheses. Comparison of large bioprostheses and mechanical prostheses populations by age groups revealed that regardless of the differences in the freedom from structural valve deterioration, the freedom from treatment failure (valve related mortality and permanent impairment from thromboembolism, anticoagulant hemorrhage, and septal emboli from prosthetic valve endocarditis) is essentially the same for mechanical prostheses and bioprostheses at 10 years. The quality of life is superior with bioprostheses, while patient survival and total valve related morbidity/mortality are similar with both types of prostheses. PMID- 8422496 TI - A five-year clinical experience with 112 Blalock-Taussig shunts. AB - In spite of recent advances in neonatal open repair for complex cyanotic heart disease, some patients require palliation with a systemic-to-pulmonary artery shunt. We report a 5-year experience (1985-1990) with 112 Blalock-Taussig shunts. Forty-six of the 92 patients had some variant of tetralogy of Fallot, with a wide spectrum of diagnoses in the remainder. The median age at surgery was 3 months. A classic Blalock-Taussig shunt was done in 26% (group I), and a 4- or 5-mm PTFE graft was utilized in the remainder (group II). The technical aspects of each of the procedures are reviewed. There were three early deaths in the entire group, none of them related to Blalock-Taussig shunt function. There was no incidence of early shunt insufficiency, bleeding, infection, limb ischemia, or pulmonary artery distortion. There was a 21% incidence of clinical congestive heart failure, seen somewhat more commonly in group I. The overall need for reshunting/open repair was similar in both groups, but there was a statistically longer interval between the initial Blalock-Taussig shunt and the second procedure in group I (21.6 vs 12.4 months). The Blalock-Taussig shunt remains a safe, reliable, and effective means of increasing pulmonary flow. PMID- 8422497 TI - Re-localization of Actsk-1 to mouse chromosome 8, a new region of homology with human chromosome 1. AB - We present here the genetic mapping of the alpha-skeletal actin locus (Actsk-1) on mouse Chromosome (Chr) 8, on the basis of the PCR analysis of a microsatellite in an interspecific backcross. Linkage and genetic distances were established for four loci by analysis of 192 (or 222) meiotic events and indicated the following gene order: (centromere)-Es-1-11.7 cM-Tat-8.3 cM-Actsk-1-0.5 cM-Aprt. Mapping of ACTSK to human Chr 1 and of TAT and APRT to human Chr 16 demonstrates the existence of a new short region of homology between mouse Chr 8 and human Chr 1. Intermingling on this scale between human and mouse chromosomal homologies that occurred during evolution creates disorders in comparative linkage studies. PMID- 8422498 TI - Characterization of four VNTR loci on human chromosome 6. AB - We have determined DNA sequences of four VNTR loci; three in the peritelomeric region of Chromosome (Chr) 6q and one at 6p21. Heterozygosities of these loci among 80 CEPH parents are 78% (D6S139), 69% (D6S149), 76% (D6S161), and 94% (D6S193), respectively. The consensus sequences of repeating units at these VNTR loci are GAGCGGCAGGGGCAGCGGGGCCTGGCCAGAGAG-(34 bp) at D6S139, CCAGGCTGGTTCACAGGCTGTGGGGTGTGATGGGTGATG (39 bp) at D6S149, GGATGGGGTTGGAGGAACTACAGAGCGGTGGTGAAGAGGA (40 bp) at D6S161, and GAGGAGGTGGGGGCCT (16 bp) at D6S193. The GC content of the consensus sequences is 62% as high as reported previously. Furthermore, we have established a PCR assay for D6S193 locus and this will be useful for individual identification or for a study of loss of heterozygosity on the long arm of Chr 6 in malignant tumors. PMID- 8422499 TI - The structure and evolution of the human salivary proline-rich protein gene family. AB - We present the nucleotide sequences of four members of the six-member human salivary proline-rich protein (PRP) gene family. The four genes are PRB1 and PRB2, which encode basic PRPs, and PRB3 and PRB4, which encode glycosylated PRPs. Each PRB gene is approximately 4.0 kb in length and contains four exons, the third of which is entirely composed of 63-bp tandem repeats and encodes the proline-rich portion of the protein products. Exon 3 contains different numbers of tandem repeats in the different PRB genes. Variation in the numbers of these repeats is also responsible for length variations in different alleles of the PRB genes. We have determined a probable evolutionary history of the human PRP gene family by comparing the nucleotide sequences of the six PRP genes. The present day six PRP loci probably evolved from a single ancestral gene by four sequential gene duplications, leading to six genes that fall into three subsets, each consisting of two genes. During this evolutionary process, multiple rearrangements and gene conversion occurred mainly in the region from the 3' end of IVS2 and the 3' end of exon 3. PMID- 8422500 TI - Genetic mapping of the athymic nude (RNU) locus in the rat to a region on chromosome 10. AB - The nude trait in the rat is transmitted in an autosomal recessive manner and is associated with thymic aplasia, T-cell deficiency, and hairlessness. Congenic rats homozygous for the RNU (Rowett nude) locus are important models in the study of inflammatory disease, tumor growth, and transplant rejection. The RNU locus has not been previously mapped, and the nature of the gene product is unknown. To determine the map location of this gene, a single F344.rnu/rnu (athymic nude congenic Fischer rat) male congenic rat was bred with 3 LEW/N (NIH stock Lewis rat) female rats to produce F1 progeny. Twelve F1 brother-sister breeding pairs were established. Forty-nine phenotypically nude F2 offspring (198 total) were obtained. Linkage analysis done on F2 DNA revealed highly significant cosegregation between the nude phenotype and eight polymorphic markers located on Chromosome (Chr) 10. The tightest linkages were with: MYH3 (embryonic, skeletal myosin heavy chain) and SHBG (sex hormone-binding globulin), giving 2 point lod scores of 20.2, and 20.0, respectively. The map order and map distances, determined by multipoint linkage calculations, were: RR24-(16.1 cM)-MYH3-(3.5 cM) SHBG-(4.7 cM)-RNU-(11.9 cM)-F16F2-(24.1 cM)-CLATP (citrate lyase ATPase)-(2.4 cM) ACE (angiotensin converting enzyme)/PPY (pancreatic polypeptide)-(14.1 cM) RR1023. The position of the RNU locus in the rat corresponds closely with that of the recently reported nu locus in the mouse. This finding suggests that the nude phenotype in the rat and the mouse arise from defects in homologous genes. PMID- 8422501 TI - Chromosomal assignment of the recoverin gene and cancer-associated retinopathy. AB - The deduced amino acid sequence of the recently cloned mouse 23kD photoreceptor cell-specific protein showed it to be identical to the recoverin protein and the CAR (cancer-associated retinopathy) protein. DNA sequence variants were found in the mouse recoverin gene (Rcvrn), and segregation analysis of restriction fragment length variants in recombinant inbred strains of mice assigned Rcvrn to mouse Chromosome (Chr) 11, between Sparc (3.7 map units) and Zfp-3 (2.3 map units). These results demonstrate a close linkage of recoverin to the tumor suppressor gene, Trp53. On the basis of these data, knowledge of the function of recoverin, and the characteristics of CAR, an experimentally testable model is presented to explain the molecular basis for CAR. PMID- 8422502 TI - Deletion mapping of the chocolate (cht) locus within the Fes-Hbb region of mouse chromosome 7. PMID- 8422503 TI - Isolation, sequencing, and expression analysis of a bovine apolipoprotein E (APOE) cDNA and chromosomal localization of the APOE locus. PMID- 8422504 TI - Mouse t haplotype-specific double insertion of B2 repetitive sequences in the Tcp 1 intron 7. PMID- 8422505 TI - Mapping of the porcine involucrin (IVL) gene to chromosome 4 by fluorescence in situ hybridization. PMID- 8422506 TI - Mapping of the porcine alpha interferon (IFNA) gene to chromosome 1 by fluorescence in situ hybridization. PMID- 8422507 TI - Mapping of the porcine SLA class I gene (PD1A) and the associated repetitive element (C11) by fluorescence in situ hybridization. PMID- 8422508 TI - Mapping of the porcine apolipoprotein B (APOB) gene to chromosome 3 by fluorescence in situ hybridization. PMID- 8422509 TI - Influence of birth weight on adult bone mineral density. AB - Bone mineral density (BMD) increases during growth until a peak is reached at maturity. The risk of development of postmenopausal osteoporosis depends on the peak bone density and the rate of its subsequent loss. To identify whether low weight at birth could affect the peak bone density, we measured BMD at both the lumbar spine and femoral neck using dual energy X-ray absorptiometry (DXA) in a group of women who had low weight at birth and in a control group of normal birth weight. There was no significant correlation between the weight at birth and the adult BMD. It appears, therefore, that low weight at birth does not influence the peak bone density and that prematurity is not a risk factor for osteoporosis. PMID- 8422510 TI - Evaluation of a program for rehabilitation of osteoporotic patients (PRO): 4-year follow-up. The Bone and Mineral Group of the University of Toronto. AB - The value of a program for the rehabilitation of osteoporotic patients (PRO) was assessed from a 4-year follow up of 139 patients referred to the program over its initial 2 years of operation. The program consisted of educational seminars, social activities and regular exercise supervision. Patients had annual clinical assessments, and bone mass measurements by neutron activation analysis (reported as CaBI). Fitness was assessed by performance on a treadmill (reported in terms of VO2max, ml/kg/min). Seventy-eight of the 139 patients remained in the program over the 4-year follow-up. This unusually high level of commitment to the program is indicative of the psychological value that patients have derived from it. The effect of the program on the osteoporosis process was inconclusive. Group 2, the 37 patients who obtained the greatest improvement in fitness (VO2max > 6 ml/kg/min), had a significantly greater reduction in back pain than did group 1, the 36 with less significant improvement (VO2max < or = 6 ml/kg/min). The bone mass and incidence of vertebral fractures on entry into the program were not significantly different between the two groups. Group 2 had on average a greater increase in bone mass over the 4 years; mean increases in CaBI, (+/- SD) were 0.09 +/- 0.09 and 0.05 +/- 0.10 for groups 2 and 1 respectively. Group 2 had on average fewer new vertebral fractures (0.08 +/- 0.36 and 0.28 +/- 0.75 for groups 2 and 1 respectively).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422511 TI - Skin color and body size as risk factors for osteoporosis. AB - We compared skin color, body size and bone mineral density (BMD) among three groups of postmenopausal women: 104 healthy black women, 45 healthy white women, and 52 osteoporotic white women with vertebral fractures. Skin color was measured by reflectometry, stature with a Harpenden stadiometer, weight with digital scales, and radial BMD by single photon absorptiometry. There were no significant differences in mean skin color (age-adjusted) between the healthy and osteoporotic white women, although both white groups differed from the black group. There was no significant correlation between skin color and BMD (age- and weight/height-adjusted) in any of the groups. All three groups differed significantly in age-adjusted BMD, although there was less difference between the healthy blacks and whites when covariates (body size, age) were taken into account. We further investigated body size differences by estimating stature at age 55 in all three groups based on our observations that osteoporotic women with vertebral fractures lose height at a rate that is 2.6 times faster than that of healthy aging women. Our analyses indicate that the osteoporotics were not shorter than the normals before the onset of their disease (based on estimated height), and do not have a significantly smaller body mass (weight/height and weight/height 2) than the normal white women. Additionally, the osteoporotics are above the ideal body mass index recommended by the National Institutes of Health. We conclude that fair skin is not a risk factor for osteoporosis and that large body size is not protective against the development of osteoporosis, although it may have a salutary effect on BMD in both blacks and whites. PMID- 8422512 TI - The effect of treatment with calcitonin on vertebral fracture rate in osteoporosis. AB - The efficacy of treatments for osteoporosis does not become evident when evaluated by fracture incidence (FI). Vertebral FI decreased in all controlled studies on calcitonin, but not significantly. Small sample sizes and short periods of treatment may have masked a possible therapeutic benefit, but longer, controlled studies with sodium fluoride or etidronate in larger groups of patients also failed to show a decrease in FI. The present analysis of nine published, therapeutic studies which indicate the FI per year and the initial prevalence of vertebral fractures, examines the question of whether the initial prevalence of fractures has an effect on the subsequent incidence of new fractures and whether the therapeutic effects have to be evaluated as a function of the initial prevalence of fractures. Bearing in mind the differences in roentgenological evaluation and in the size and quality of the various studies, the analysis revealed (1) that in the control groups there was a higher FI in patients with more than three vertebral fractures at baseline (estimated odds ratio (OR) = 49, p = 0.011); (2) that a similar trend, although not statistically significant, was observed in treated patients; (3) that the groups of control patients treated for more than 1 year showed in general an increase in FI beyond the first year and that the reverse was true in treated patients. In conclusion, failure to allow for the initial prevalence of vertebral fractures at the individual level in therapeutic trials of calcitonin to treat osteoporosis and prevent new fractures might have contributed to the absence of a demonstrable benefit of the treatment in those studies. PMID- 8422514 TI - Ultrasonic velocity measurements through the calcaneus: which velocity should be measured? AB - The assessment of skeletal integrity by the measurement of ultrasonic velocity through the calcaneus has only recently become widely available and is usually made in conjunction with the measurement of broadband ultrasonic attenuation. Using data obtained with a contact ultrasonic bone analyser (CUBA) system, this report examines whether ultrasonic studies of the heel require the measurement of true velocity of sound in the calcaneus (Vbone), or whether heel velocity (Vheel, defined as the mean velocity through bone and soft tissue) or time of flight velocity (Vtof, defined as the mean velocity between the two transducers) are adequate surrogates. The populations selected for study were 15 healthy young women (group 1, mean age 26 years), 231 healthy peri- and postmenopausal women (group 2, mean age 52 years) and 33 osteoporotic women with confirmed vertebral fracture (group 3, mean age 66 years). Precision was studied by performing 10 repeated scans on the subjects in group 1 and duplicate scans on 144 women randomly selected in groups 2 and 3. Precision was expressed as the percentage coefficient of variation (CV). Both precision studies yielded similar results. The precisions (and 5% to 95% ranges) for all groups combined were: Vbone 2.71% (1465-1809 m/s); Vheel, 1.10% (1511-1646 m/s): Vtof, 0.70% (1349-1425 m/s). Although the precision data suggest Vtof should be preferred, when the range of clinical values is taken into account the smaller CV is exactly cancelled by the narrower range.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422513 TI - Loss of bone mass in patients with Klinefelter's syndrome despite sufficient testosterone replacement. AB - To determine whether testosterone replacement therapy reverses the detrimental effects of hypogonadism on bone density, we measured the total body, lumbar spine and proximal femur bone mineral density (BMD) by dual-energy X-ray absorptiometry in 14 patients with Klinefelter's syndrome on long-term testosterone replacement therapy and compared the results with 14 age- and sex-matched normal controls. Seven of the patients were receiving oral testosterone undecanoate thrice daily (240 mg/day) and the others were having intramuscular testosterone enanthate injections once every 3 weeks (250 mg/injection). Their serum testosterone levels were maintained within the normal limits (10-40 nmol/l). We showed that patients on testosterone replacement had decreased amount of bone density in the left femoral neck when compared with the controls (p < 0.01). Similar decreases were also observed in the left Ward's triangle (p < 0.01) and in the left trochanter (p < 0.05). There were no significant differences in the total body and the lumbar spine measurements in these two groups of subjects. No correlation was found between the BMD values of femur and the duration of testosterone treatment in the patients with Klinefelter's syndrome. The type of testosterone treatment was also not associated with significant differences in BMD. In conclusion, sufficient testosterone replacement with currently available methods does not reverse the decrease in bone mass associated with hypogonadism in patients with Klinefelter's syndrome. PMID- 8422515 TI - Cross-calibration of liquid and solid QCT calibration standards: corrections to the UCSF normative data. AB - Quantitative computed tomography (QCT) has been shown to be a precise and sensitive method for evaluating spinal bone mineral density (BMD) and skeletal response to aging and therapy. Precise and accurate determination of BMD using QCT requires a calibration standard to compensate for and reduce the effects of beam-hardening artifacts and scanner drift. The first standards were based on dipotassium hydrogen phosphate (K2HPO4) solutions. Recently, several manufacturers have developed stable solid calibration standards based on calcium hydroxyapatite (CHA) in water-equivalent plastic. Due to differences in attenuating properties of the liquid and solid standards, the calibrated BMD values obtained with each system do not agree. In order to compare and interpret the results obtained on both systems, cross-calibration measurements were performed in phantoms and patients using the University of California San Francisco (UCSF) liquid standard and the Image Analysis (IA) solid standard on the UCSF GE 9800 CT scanner. From the phantom measurements, a highly linear relationship was found between the liquid- and solid-calibrated BMD values. No influence on the cross-calibration due to simulated variations in body size or vertebral fat content was seen, though a significant difference in the cross calibration was observed between scans acquired at 80 and 140 kVp. From the patient measurements, a linear relationship between the liquid (UCSF) and solid (IA) calibrated values was derived for GE 9800 CT scanners at 80 kVp (IA = [1.15 x UCSF] - 7.32).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422516 TI - Fractures of the proximal femur in Picardy, France, in 1987. AB - Between 1 January and 31 December 1987, 1178 hip fractures were recorded in the 28 clinical centres, public and private, of the Picardy region (19443 km2, 1.8 million inhabitants). Patients under 20 years and those with metastatic cancer and myelomatous fractures were excluded. Women sustained 853 fractures (age mean +/- SD: 80.2 +/- 10.4 years) and men 325 (age 69.7 +/- 16.0 years). The crude incidence rate per 10,000 person years was 13.4 for women and 5.4 for men (female/male ratio 2.6). These incidences are among the lowest recorded in Northern Europe. Women with trochanteric fractures were older than those with cervical ones, but no difference was observed for men. After adjusting for age and sex, the incidence of hip fracture was greater in urban (10.5 per 10,000 person years) and semi-rural areas (8.2) than in rural areas (5.3). The mean bed days per patient (+/- SD) was 21.6 +/- 16.0 (quartiles: 13-17-26 days); no difference was observed between sex or age classes. The in-hospital mortality rate was 8.7%, it increased with age and was higher in men, whatever their age. We review the data in different countries, mostly European, to compare with the Picardy region. PMID- 8422517 TI - The effect of the menopause and hormone replacement therapy on serum carboxyterminal propeptide of type I collagen. AB - We investigated the effect of the menopause and postmenopausal hormone replacement therapy on the serum concentration of carboxyterminal propeptide of type I procollagen (PICP), which is a biochemical marker of type I collagen synthesis. A group of 124 healthy postmenopausal women, aged 45-53 years, had about 20% higher serum PICP than did a group of 40 healthy premenopausal women aged 35-52 years (114 +/- 35 micrograms/l vs. 95 +/- 26 micrograms/l (mean +/- SD); p = 0.002). The 124 postmenopausal women were also participating in a double masked longitudinal study with two placebo groups and four different hormone replacement therapy groups. The four hormone regimens resulted in similar responses in serum PICP. Compared with placebo, 1 year of treatment with any of the four hormone replacement therapies significantly decreased serum PICP to premenopausal levels. We conclude that the formation of type I collagen is increased shortly after the menopause and that hormone replacement therapy reverses this increase. PMID- 8422518 TI - Muscle strength in osteoporotic versus normal women. AB - Strong back muscles contribute to good posture and skeletal support. Osteoporosis, being a metabolic bone disease, should not affect muscle strength. In this study we were interested in comparing the back extensor strength (BES) of osteoporotic and normal women. Fifty-five women ages 40-85 years who had a documented diagnosis of osteoporosis and were referred for initiation of proper exercise programs were included in our study after meeting the inclusion criteria. They all had evaluation of their posture, back and upper extremity strength, and physical activity score through our Rehabilitation of Osteoporosis Program--Exercise (ROPE). In addition, to avoid the interference of pain on application of maximal effort, we did not include subjects with acute back pain or those who experienced back pain with maximal effort during the testing trial. BES for osteoporotic women ranged from 16 to 65 lb (mean +/- SD, 36.5 +/- 15.5) for ages 40-59 years, 9 to 55 lb (mean +/- SD, 29.9 +/- 10.6) for ages 60-69 years, 6 to 52 lb (mean +/- SD, 24.3 +/- 10.2) for ages 70-79 years, and 17 to 27 lb (mean +/- SD, 21.2 +/- 4.2) for ages 80 years or older. Comparison of these data with the BES of 25 normal women, with statistical adjustment for age, demonstrated that the osteoporotic women had significantly lower BES than the normal women. A longitudinal study of a larger group of women would be of great interest for clarifying whether the weakness of back extensors precedes and, indeed, contributes to compression fractures of the spine. PMID- 8422519 TI - Antibiotic substance produced by a newly isolated marine microalga, Chlorococcum HS-101. PMID- 8422520 TI - Comparison of Artemia feeding regimens on larval growth in a short-term fathead minnow toxicity test. PMID- 8422521 TI - Copper toxicity and phosphate utilization in the cyanobacterium Nostoc calcicola. PMID- 8422522 TI - Cytotoxic effects of distillary waste on Allium cepa L. PMID- 8422523 TI - Selective protection of temperature against cadmium acute toxicity to Bufo arenarum tadpoles. PMID- 8422524 TI - An on-site immunoassay for detecting PCB in soil. PMID- 8422525 TI - Degradation of dacthal and its metabolites in soil. PMID- 8422526 TI - Heavy metals in epigeic fauna: trophic-level and physiological hypotheses. PMID- 8422527 TI - Concentration and bioavailability of heavy metals in sediments in Lake Yojoa (Honduras). PMID- 8422528 TI - Survey of chemical (mostly metals) poisoning cases as reflected in hospital admissions in urban Zimbabwe. PMID- 8422529 TI - Evaluation of acute immunotoxicity of alachlor in male F344/N rats. PMID- 8422531 TI - Toxicological impacts of benzophenone on the liver of guinea pig (Cavia porcellus). PMID- 8422530 TI - Relationship among cyanide-induced encephalopathy, blood ammonia levels, and brain aromatic amino acid levels in rats. PMID- 8422532 TI - Age and sex related behavioral changes induced by dibutyltin-dilaurate in rats. PMID- 8422533 TI - Effect of sibship on DDT residue levels in human serum from a malaria endemic area in northern Kwazulu. PMID- 8422534 TI - Organochlorine insecticide and polychlorinated biphenyl residues in human breast milk in Madrid (Spain). PMID- 8422535 TI - Levels of chlordane, oxychlordane, and nonachlor on human skin and in human blood. PMID- 8422536 TI - The major lines of metazoan evolution: summary of traditional evidence and lessons from ribosomal RNA sequence analysis. PMID- 8422537 TI - Molecular biology of excitatory amino acid receptors: subtypes and subunits. AB - Glutamate receptors coupled to ion channels have been named according to their selective agonist: N-methyl-D-Aspartate (NMDA), kainate, quisqualate and alpha amino-3-hydroxy-5-methyl-4-isoxazole-propionate (AMPA). The pharmacology of the NMDA receptor is clearly different from that of the kainate, quisqualate and AMPA receptors, thus differentiating two types: NMDA and non-NMDA receptors. Molecular cloning and expression of non-NMDA receptor subunits have now established that the different neuronal responses to kainate, quisqualate and AMPA are mediated by at least two subtypes of ligand-gated channels: one responding to the three ligands, the other responding to kainate and quisqualate but not to AMPA. PMID- 8422538 TI - Structural and functional conservation of serotonin receptors throughout evolution. AB - Pharmacological studies as well as molecular cloning of serotonin receptors have revealed a multiplicity of receptor subtypes not only in mammals but also in molluscs and arthropods. Most of these receptors belong to the G protein-coupled receptor family and modulate levels of second messengers such as cAMP, IP3 and calcium. Rather than being specialized in a particular physiological function, a given receptor may be expressed in multiple neurons throughout the brain but always in the same compartment of these neurons. The 5HT1B receptor for example, is generally localized presynaptically on neuronal terminals where it inhibits neurotransmitter release. A widespread distribution of 5-HT receptors might explain how serotonin can modulate the multiple neuronal circuits which underlie complex behaviors. PMID- 8422539 TI - The wide range of actions of the FMRFamide-related peptides and the biological importance of peptidergic messengers. AB - The importance of peptides as intercellular messengers is discussed. The view is put forward that peptides evolved early in evolution as chemical messengers and that they have come to exert a wide range of actions. Using as an example the FMRFamide (Phe-Met-Arg-Phe-NH2) related peptide family of molluscs, the wide range of peptide actions on membrane currents is discussed and considered in relation to co-localization of peptides with low molecular weight (or "classical") intercellular messengers. PMID- 8422540 TI - Molecular studies on insect octopamine receptors. AB - Octopamine receptors are widely distributed in the insect nervous system and carry out a range of functions equivalent to the adrenergic receptors of the vertebrate sympathetic nervous system. Molecular studies on insect octopamine receptors have concentrated upon molecular pharmacological approaches to identify the particular subtype of octopamine receptor mediating its effects in a particular tissue and on the modes of action of the receptors in a particular tissue. Molecular biological approaches are now being pursued to define the structure of the octopamine receptor. Recent findings in this area will be reviewed, along with promising approaches for future molecular studies on insect octopamine receptors. PMID- 8422541 TI - Acetylcholine receptor molecules of the nematode Caenorhabditis elegans. AB - Receptors for acetylcholine are present in nematodes. Studies using physiological and biochemical methods have revealed the existence of nicotinic acetylcholine receptors with a novel pharmacology. Caenorhabditis elegans provides a particularly suitable organism with which to investigate such receptors using molecular genetic approaches. Mutants resistant to the cholinergic agonist (and anthelmintic drug) levamisole have permitted the isolation of a number of genes, including structural subunits of the nicotinic acetylcholine receptor. The only known viable mutants of nicotinic receptors are those of Caenorhabditis elegans. This organism offers the prospect of studying the developmental and regulatory effects of the loss of a single component of the receptor. Using Caenorhabditis elegans it is possible to select interesting phenotypic mutations by in vivo mutagenesis before determining the causative lesion. Resistance genes other than those encoding structural subunits are of particular interest, as they will encode additional polypeptides closely associated with nicotinic receptor function. Such proteins are often difficult or impossible to identify using conventional biochemical approaches, whereas genetic selection should permit their identification. PMID- 8422542 TI - Muscarinic acetylcholine receptors in invertebrates: comparisons with homologous receptors from vertebrates. AB - The pharmacology, physiology and molecular biology of invertebrate muscarinic acetylcholine receptors are compared with current knowledge concerning vertebrate muscarinic acetylcholine receptors. Evidence for the existence of multiple receptor subtypes in invertebrates is examined, emphasizing what is presently known about the sensitivity of invertebrate preparations to subtype selective ligands previously defined in vertebrate studies. Other evidence for muscarinic receptor subtypes which is examined includes: heterogeneous responses to classical muscarinic ligands and evidence for coupling of invertebrate muscarinic receptors to several different classes of second messenger systems. Clues regarding possible functions for invertebrate muscarinic receptors are discussed, including evidence from both physiological studies and in situ localization studies which reveal patterns of receptor protein and mRNA expression. A detailed analysis of the structural similarities between a cloned Drosophila muscarinic receptor and vertebrate muscarinic receptors is also presented. Regions of the receptors that may be involved in ligand binding, effector coupling and receptor regulation are identified in this comparison. Future directions for invertebrate muscarinic receptor research are considered including: methods for cloning other receptor subtypes, methods for cloning homologous receptors from other species and genetic approaches for determining the physiological roles of muscarinic receptors. PMID- 8422543 TI - Molecular characteristics of HGF and the gene, and its biochemical aspects. AB - Hepatocyte growth factor (HGF) was initially identified as a potent mitogen for mature hepatocytes in sera of partially hepatectomized rats and seems to be the long sought after hepatotrophic factor for liver regeneration. HGF, first purified to homogeneity from rat platelets in 1986, is a disulfide-linked heterodimer molecule composed of a 69 kDa alpha-subunit and a 34 kDa beta subunit. In 1989, cDNAs of both human and rat HGF were cloned and the primary structure of HGF was determined. This factor is derived from a single chain precursor of 728 amino acid residues, which is proteolytically processed to form mature HGF. While HGF has no homology wit any known growth factors, it does have a considerable homology (38%) with plasminogen. The alpha-subunit of HGF contains four kringle domains and the beta-subunit resembles serine proteases, but there is no evidence of protease activity. The human HGF-encoding gene is composed of 18 exons and 17 introns, spans about 70 kb and located on chromosome 7q11.1-21 in humans. The overall gene organization of the HGF gene is highly homologous with that of plasminogen. These genes seem to be derived from a common ancestor, by gene duplication. Artificially mutated HGFs with deletion of the N-terminal hairpin structure or the first or second kringle domain lose almost completely all biological activities, hence these domains are probably essential for the biological activities of HGF and possibly for binding to its receptor. A high affinity receptor for HGF with a Kd value of 20-30 pM was detected on various epithelial cells, including hepatocytes. The c-met proto-oncogene product was recently identified to be an HGF receptor. Transfection of c-met cDNA into COS cells provided further evidence that the c-met product is a functional high affinity receptor for HGF capable of transducing the mitogenic signal of HGF. PMID- 8422544 TI - In vitro regulation of HGF-SF expression by epithelial-mesenchymal interactions. AB - Studies of parameters which affect cellular proliferation, cellular differentiation together with studies of cell-cell interactions which affect cell behavior are particularly interesting in that they can be used to identify and characterise molecules which through changes in gene expression induce/inhibit cell proliferation, differentiation and movement. Such studies are crucial not only in the context of understanding growth and development, but also in understanding the processes of wound healing and regeneration as well as tumor invasion and metastasis. Here we present a summary of some cell culture models which we have developed for the study of the above-mentioned phenomenon, together with their application to the studies of regulation of HGF-SF expression. PMID- 8422545 TI - Roles of HGF as a pleiotropic factor in organ regeneration. AB - Hepatocyte growth factor (HGF) has been discovered, purified, and molecularly cloned as a potent mitogen for mature hepatocytes. HGF is produced by mesenchymal cells and predominantly acts on a wide variety of epithelial cells, as a mitogen (stimulation of cell growth), a motogen (stimulation of cell motility), and a morphogen (induction of multicellular architecture). HGF mRNA and HGF protein are rapidly and markedly increased in the liver and plasma of rats with various types of liver injuries, and HGF receptors on plasma membranes of the liver were almost completely down-regulated due to HGF-binding and subsequent internalization. Thus HGF acts as a hepatotrophic factor for regeneration; intravenously injected recombinant HGF remarkably enhances liver regeneration. HGF mRNA, HGF protein, and HGF receptors in the kidney are also rapidly altered prior to the onset of renal tubular cell replication after various renal insults, thus HGF also acts as a renotropic factor in renal regeneration. HGF mRNA is rapidly induced not only in the injured organ but also in non-injured organs such as the lung and spleen following hepatic or renal injury. Because "injurin", a protenous factor which induces HGF mRNA expression, was detected in the plasma of rats with various organ injuries, it seems to increase HGF mRNA expression in non-injured organs and HGF produced in non-injured organs may also participate in the regeneration of the liver or kidney through an endocrine mechanism. Based on these results and its uniquely suited ability to act as mitogen, motogen and morphogen, HGF is considered to be a key molecule for construction of normal tissue structure during embryogenesis, organogenesis, and organ regeneration. PMID- 8422546 TI - The role of HGF-SF in animal and human hepatic physiology and pathology. AB - Hepatocyte growth factor (HGF) was first described as a hepatotrophic factor in partially hepatectomized rat plasma in the early 1980's and was purified from plasma of a patient with fulminant hepatic failure and from rat platelets in 1986 1987. Recent progress has revealed that HGF is the same protein as scatter factor and tumor cytotoxic factor, and is now known to be a broad-spectrum growth factor which stimulates cell growth not only of hepatocytes but also of many other types of epithelial and endothelial cells. In this review, however, we concentrate on the role of HGF, mainly human HGF, on liver regeneration after injury. In humans, plasma levels of hHGF increase to greater than 10 ng/ml during severe liver disease such as fulminant hepatic failure and decrease rapidly to normal levels when the patients recover from the disease. In less severe liver damage such as occurs in acute hepatitis, levels of hHGF in plasma increase to 0.5-1 ng/ml which is approaching the half maximal concentration for the stimulation of DNA synthesis in human hepatocytes in culture. Thus, HGF is believed to be involved in control of liver regeneration. Although the cell type(s) which produces HGF during liver disease is not yet identified, it is possible that circulating leukocytes or splenocytes are responsible. Synthesis of HGF is though to be regulated by a putative inducer(s) derived from damaged liver tissue. A control mechanism for HGF production during liver disease is proposed. PMID- 8422547 TI - Metabolism of HGF-SF and its role in liver regeneration. AB - HGF-SF is rapidly sequestered in the liver after intravenous injection. This demonstrates that liver is the major organ responsible for clearance of HGF-SF. A minor involvement of the kidney in that function was also shown. Plasma levels of HGF-SF mRNA increase by 15-17 fold rapidly after 2/3 partial hepatectomy. This increase precedes the increase of HGF-SF mRNA in liver following surgical resection. The findings strongly suggest that the observed rapid elevation in plasma HGF-SF after 2/3 partial hepatectomy is due to decrease of clearance by the liver. A mechanism is proposed whereby changes in HGF-SF levels due to removal of hepatic mass stimulate entry of hepatocytes from G0 to G1 and eventually lead to DNA synthesis. PMID- 8422548 TI - Properties and structure-function relationship of HGF-SF. AB - HGF-SF is a cytokine independently isolated and characterized for its activities as a mitogen for liver cells in culture (hepatocyte growth factor, HGF) and as a factor which enhances the movement and induces the dissociation of epithelial colonies (scatter factor, SF). The factor is composed of two subunits (A and B) with a M(r) of approximately 57 K and approximately 30 K produced by proteolytic cleavage of a polypeptide precursor. Based on the complete conservation of critical cysteine residues, the domain structure of HGF-SF appears to be remarkably similar to the domain structures of plasminogen, (a blood protease involved in fibrinolysis) and the HGF-like protein (the translation product of a gene recently isolated and which shares extensive sequence identity with HGF-SF). The A subunit of HGF-SF consists of an N-terminal domain, homologous to plasminogen preactivation peptide (PAP), followed by four kringle domains. The B subunit has the basic structure of a serine protease domain although HGF-SF has no protease activity due to replacement substitutions in two of the three residues of the catalytic site. Experiments with a truncated form of HGF-SF generated by alternative splicing and with a series of deletion and point mutants generated by site-directed mutagenesis, have established that both the N-terminal and the kringle domains of the A chain and the B chain are all required for biological activity. Loss of mitogenic activity in the HGF-SF mutants is always associated with loss of motogenic activity and vice versa. Thus, the protein engineering experiments confirm the hypothesis that the mitogenic and motogenic responses to HGF-SF diverge at the post-receptor level. PMID- 8422549 TI - Gene expression and regulation of HGF-SF. AB - Molecular cloning and characterization of cDNAs and genomic DNA for human HGF-SF revealed the expression pattern of the gene. Alternative use of splicing sites and processing/polyadenylation sites generates multiple mRNAs for human HGF-SF in a variety of tissues and cell lines. This alternative mRNA production may be regulated in a tissue-specific manner. Full-length HGF-SF is encoded by the 6.3 kb or 3.1 kb mRNA. A variant form of HGF-SF is encoded by the 1.5 kb mRNA which is generated by alternative splicing accompanied by the utilization of an alternative processing/polyadenylation site in an extra exon. The variant form consists of an N-terminal sequence and the first two kringles and acts as an antagonist of HGF-SF mitogenic activity. HGF-SF with a deletion of 5 amino acids in the first kringle is produced from an alternatively spliced mRNA. The deletion induces the change in the heparin-binding property of the protein. Analysis of responses of the HGF-SF mRNA during liver regeneration revealed that the HGF-SF gene is activated. The level of mRNA markedly increases in the rat liver, spleen and lung after administration of hepatotoxins. Characteristic regulatory elements, an IL6 response element and an NF-IL6 binding element, are present proximal to the major transcription initiation site in the human HGF-SF gene. These elements may be involved in the activation of the gene after liver injury. The potential autocrine role of HGF-SF in the acquisition of altered phenotypes was determined by introduction of the gene into target cells for HGF-SF. Autocrine production of the factor induces changes in cell properties from parental types to those obtained by the addition of exogenous factor. PMID- 8422550 TI - Genomic organization, chromosomal localization and developmental expression of hepatocyte growth factor-like protein. AB - Hepatocyte growth factor is a multi-functional protein that elicits different biological responses in a tissue- or cell-specific manner. We have isolated the cDNA and gene for a previously unidentified protein that has the identical domain composition as hepatocyte growth factor. We have called this novel protein "hepatocyte growth factor-like protein". Both proteins contain four kringle domains followed by a serine protease-like domain. Overall, the two proteins are only about 50% identical. The human gene for hepatocyte growth factor-like protein has been localized to locus p21 on chromosome 3, where one or more tumor suppressor genes may be located. Although the biological function of HGF-like protein has not been determined we have localized the major site of synthesis to the liver. Whether this newly identified member of the hepatocyte growth factor family of proteins is involved in the development and differentiation of the liver remains to be determined. PMID- 8422551 TI - Electrotherapy. PMID- 8422552 TI - Systemic lupus erythematosus and genes within the HLA region. PMID- 8422553 TI - The stiffness of normal articular cartilage and the predominant acting stress levels: implications for the aetiology of osteoarthrosis. AB - A survey was conducted of compressive stiffness of normal articular cartilage over the entire surfaces of 13 knees and 10 ankle joints, using an indentation technique. Contoured maps of the stiffness of these joints was obtained. A direct relationship found between the stiffness and the predominant level of stress to which the cartilage was subjected, suggests a process of cartilage adaptation to stress. This finding, together with further evidence of osteoarthrotic damage in areas where infrequent but excessive levels of stress occur, supports a hypothesis which implicates a particular pattern of mechanical stress as being an important aetiological factor in osteoarthrosis. PMID- 8422554 TI - Reflex sympathetic dystrophy (algodystrophy) PMID- 8422555 TI - Prevention of collagen induced arthritis in mice by deletion of T cell receptor V beta 8 bearing T cells with monoclonal antibodies. AB - Collagen induced arthritis (CIA) is an animal model of inflammatory polyarthritis. Immunotherapy with the monoclonal antibody F23.1, which deletes V beta 8 bearing T cells, significantly decreased the incidence of CIA in mice. Treatment with the monoclonal antibody 466B5, to delete V beta 6 bearing T cells in combination with F23.1 was no more effective than F23.1 alone and the CIA incidence in 466B5 treated animals was not significantly different from controls. Thus, the V beta 8 family of T cell receptor is expressed on self-reactive T cells in the CIA model of B10.RIII mice injected with porcine type II collagen. PMID- 8422556 TI - Thyroid disease and other autoimmune phenomena in a family study of primary Sjogren's syndrome. AB - Autoimmune diseases and autoantibodies have been documented in 42 index cases with definite primary Sjogren's syndrome (1 degree SS), 207 relatives and 39 spouses. The results were compared with control data from a local population survey. Thyroid disease, 1 degree SS and their associated autoantibodies were the commonest autoimmune abnormalities observed and found predominantly in older female relatives. The HLA-DR3 phenotype associated with 1 degree SS, antinuclear factor, hypothyroidism, and thyroid microsomal antibody. Rheumatoid arthritis and systemic lupus erythematosus were not found in excess in the families. Primary Sjogren's syndrome is frequently associated with thyroid disease and we suggest that there is a common genetic predisposition between these diseases which differs from 2 degrees SS associated with rheumatoid arthritis and systemic lupus erythematosus. This includes MHC and non-MHC genes. PMID- 8422557 TI - The SCID mouse as a vehicle to study autoimmunity. AB - Peripheral blood mononuclear cells (PBMC) injected into severe combined immune deficiency (SCID) mice continue to secrete human immunoglobulin and respond to immunization with recall antigens. PBMC for patients with autoimmune diseases produce autoantibodies of the same specificities but at lower levels compared to the donor. SCID recipients of patients' PBMC fail to develop clinical disease although some histological lesions suggestive of autoimmunity have been reported. Transfer of autoimmunity from rodents to SCID mice has been successful in some instances. Despite obstacles related to limited survival and varying degrees of graft vs host disease (GVHD), SCID mice should prove to be a useful vehicle to explore autoantibody regulation. PMID- 8422558 TI - Quantitative bone scintigraphy in reflex sympathetic dystrophy. AB - We have investigated the appearances of the technetium-99m labelled methylene diphosphonate bone scan in 29 scans performed in 16 patients with post-fracture reflex sympathetic dystrophy (RSD) of the hand. There was a close correlation between increased uptake in the metacarpophalangeal joints and the metacarpal bones, suggesting that the increased uptake in RSD is not confined to the periarticular areas as has been previously reported but occurs throughout the affected region. In seven cases examined 3 months after fracture, RSD was associated with a significant increase in uptake at all sites in the hand on the delayed scan which was not seen in matched post-fracture controls, confirming that increased uptake on delayed bone scintography is a sensitive test for the presence of RSD even in the presence of a fracture of the wrist. The early increased uptake gradually returned to normal. Local tenderness assessed by dolorimetry was correlated with increased uptake, suggesting that the bone scan is not only useful as a diagnostic tool but also provides a quantitative indication of the severity of the condition which may be useful in planning and assessing treatments. PMID- 8422559 TI - Changes in cortical and trabecular bone in algodystrophy. AB - We studied the pattern of bone loss in the hand of 77 patients with Colles' fracture using metacarpal morphometry, single photon absorptiometry and a radiographic scoring system. Forty-four patients had post-traumatic algodystrophy and the remainder served as controls. Both groups were immobilized in the same manner and for the same period of time and both showed loss of bone during immobilization. The loss of bone 7 weeks after fracture was significantly greater in algodystrophy than in controls both at cortical (P < 0.05) and at trabecular sites (P < 0.001). Recovery of bone occurred in the control patients by 19 weeks after fracture at cortical sites and by 31 weeks in trabecular bone. In contrast, the bone loss seen in patients with algodystrophy persisted for the 6-month duration of the follow-up, and up to 1 year in all nine patients studied for longer. These findings indicate that post-traumatic algodystrophy is associated with regional skeletal losses greater than those following uncomplicated fracture and may result in irreversible changes in the structure and thus the strength of the bony architecture. PMID- 8422560 TI - A preliminary study on the interferon-alpha treatment for xerostomia of Sjogren's syndrome. AB - We studied the therapeutic effect of interferon-alpha on the xerostomia of Sjogren's syndrome by injecting 1 x 10(6) units of interferon-alpha intramuscularly once weekly. Saliva production was quantitated by the Saxon test. Variation of saliva production measured at monthly intervals during the 3-month period prior to administration of interferon-alpha was within +/- 0.30 g/2 min. After administration of interferon-alpha, saliva production increased to greater than 0.30 g/2 min in six patients, and the increase was statistically significant by the paired t-test (P = 0.002). The result suggests a beneficial effect of this agent in increasing the saliva production of patients with primary Sjogren's syndrome. PMID- 8422561 TI - Chlormezanone in primary fibromyalgia syndrome: a double blind placebo controlled study. AB - Primary fibromyalgia syndrome (PFS) is a common condition that often proves resistant to health interventions. Chlormezanone combines corrective effects on disturbed sleep with muscle-relaxant properties, and therefore could be of potential benefit in PFS. Forty-two female patients with PFS (mean age 49, range 24-72 years) were randomly and blindly allocated either chlormezanone 400 mg nocte or placebo. Patients were assessed by single observer at 0, 3 and 6 weeks of treatment; assessments included sleep quality, inactivity and morning stiffness, morning alertness, tender point score, mood change and global opinion (patient and observer). No beneficial therapeutic effect could be attributed to chlormezanone. Although there are problems in assessing severity of a predominantly subjective condition, this essentially negative finding is of interest in respect to the pathogenesis of PFS. PMID- 8422562 TI - Carboxyterminal type I procollagen peptide concentrations in systemic sclerosis: higher levels in early diffuse disease. AB - Systemic sclerosis (SSc) is characterized by increased collagen deposition in skin and internal organs, and increased serum concentrations of different connective tissue metabolites have been reported. In this study serum concentrations of carboxyterminal type I (PICP) and aminoterminal type III procollagen peptide (PIIINP) were higher in 24 patients with diffuse cutaneous systemic sclerosis (dSSc), than in 30 patients with limited cutaneous systemic sclerosis (lSSc). Thirty patients with advanced and progressing disease had higher serum concentrations of both peptides than 24 patients with milder disease. In patients with advanced disease, both peptides showed a negative correlation to disease duration. Despite being relatively higher both in early, widespread and advanced disease, serum PICP concentrations varied only within the normal range for healthy controls. Thus PICP is of limited value as a marker of disease activity in SSc. PMID- 8422563 TI - Salmonella bacteraemia occurring concurrently with the first presentation of systemic lupus erythematosus. AB - There is increasing recognition of the association between Salmonella infections and systemic lupus erythematosus (SLE). However, these infections have usually occurred in patients with established SLE who were receiving immunosuppressive therapy. We describe two young women with previously undiagnosed SLE who developed Salmonella bacteraemia. Neither of these patients had received immunosuppressive drugs. From these and two previously reported patients, it may be concluded that SLE per se is likely to be associated with a defect in host defences to Salmonella bacteria. PMID- 8422564 TI - Gastro-duodenal damage due to non-steroidal anti-inflammatory drugs in children. AB - Thirteen juvenile chronic arthritis patients with abdominal symptoms related to non-steroidal anti-inflammatory drug therapy were endoscoped before and after a 6 week course of either misoprostol or ranitidine therapy. Major presenting symptoms were generalized abdominal pain and nausea. Symptoms did not correlate well with endoscopic findings which revealed no evidence of ulceration and minimal erosive damage. Five patients had mild erythema or gastritis. Bleeding lesions were confined to small numbers of petechiae. Following treatment with either misoprostol or ranitidine, patients improved symptomatically without a corresponding improvement on endoscopic and histological examination of stomach and duodenum. Both treatments were well tolerated. PMID- 8422565 TI - The use of non-steroidal anti-inflammatory drugs in paediatric rheumatic diseases. AB - The use of NSAIDs for arthritis differs in children from adults in their indications, uses and pharmacokinetics, and fewer are available. Children with arthritis are assessed differently, as they complain less of pain. Salicylates, indomethacin and ibuprofen are used for the fever of systemic JCA. For control of joint symptoms, diclofenac, ibuprofen, tolmetin and naproxen are equal in their efficacy and tolerance:salicylates and indomethacin are no more effective but more toxic. Children tolerate NSAIDs well. Gastrointestinal symptoms appear to be less common than in adults, but the evidence regarding endoscopic changes in conflicting. Renal toxicity is rare. Tolmetin can cause pseudoproteinuria and naproxen pseudoporphyria. The liver in systemic JCA is vulnerable to drug toxicity. A therapeutic trial of an NSAID should continue for 8 weeks. Interactions with methotrexate and carbonic anhydrase inhibitors for glaucoma complicating iridocyclitis may occur. PMID- 8422566 TI - Systemic-onset juvenile chronic arthritis and bone marrow hypoplasia. AB - A two- and-a-half-year-old girls with systemic-onset JCA developed pancytopaenia 21 days after her illness began. Bone marrow examination revealed hypoplasia, with no evidence of erythrophagocytosis. Two weeks later peripheral blood specimens showed evidence of bone marrow regeneration. No definite cause for the hypoplasia was found subsequently. Persistent haematological abnormalities after resolution of the hypoplasia included anaemia of chronic inflammation, leucocytosis and thrombocytosis. Unexplained bone marrow hypoplasia has not previously been described in systemic-onset JCA. PMID- 8422567 TI - Special interest group for joint hypermobility. PMID- 8422568 TI - Arthritis and Rheumatism Council funding priorities. PMID- 8422569 TI - The coexistence of intractable nodular rheumatoid arthritis and scleroderma. PMID- 8422570 TI - Streptococcal arthritis and erysipelas complicating dental extraction. PMID- 8422571 TI - Vasculitis, Raynaud's phenomenon and polyarthritis associated with gemfibrozil therapy. PMID- 8422572 TI - Is the hip involved in generalized osteoarthritis? PMID- 8422573 TI - Digital vasculopathy associated with hyperparathyroidism. PMID- 8422574 TI - The monitoring of patients on disease-modifying antirheumatoid drugs. PMID- 8422575 TI - Vasculitis associated with herbal preparation containing Passiflora extract. PMID- 8422576 TI - Salmonella septic arthritis in HIV patients. PMID- 8422577 TI - Expression of gamma-aminobutyric acid immunoreactivity in reactive astrocytes after ischemia-induced injury in the adult forebrain. AB - Transient ischemia induces an increase in glial fibrillary acidic protein (GFAP) immunoreactivity which can be detected in specific forebrain regions of the adult gerbil as early as day 2, becomes prominent by day 4-7 and persists for at least 3 months. These forebrain areas include layers 2/3 of the somatosensory and auditory cortices, the CA1 and CA4 sectors of the hippocampus, the dorsolateral region of the striatum, and the dorsolateral subregion of the medial septal nucleus. In addition, astrocytes in the ischemically lesioned areas stain with gamma aminobutyric acid (GABA) antiserum. These GABA-immunoreactive astrocytes are not found in non-damaged areas. The time-course of expression of GABA immunoreactivity is similar to that of GFAP immunoreactivity. Using a double immunofluorescent staining method, reactive astrocytes which express GABA immunoreactivity were also found to immunostain with either GFAP or vimentin. On the other hand, astrocytes were not found to be immunoreactive with antibodies to glutamic acid decarboxylase or glutamate. Our present finding demonstrates, in an in vivo model, an aberrant expression of GABA immunoreactivity by astrocytes which is not observed in non-ischemic adult animals. PMID- 8422578 TI - Thermal dependence of chemosensory activity in the carotid body superfused in vitro. AB - We studied the relationship between chemosensory activity and temperature in carotid bodies excised from pentobarbitone-anesthetized cats, and superfused in vitro at flows between 0.4 and 2.0 ml/min with modified Tyrode's solution buffered with HEPES at pH 7.43. The basal frequencies of chemosensory discharges were recorded from the entire carotid nerve at different steady thermal conditions. For preparations superfused with saline equilibrated with 100% O2, thermally dependent increases in frequency were observed, with significant differences between all nearby thermal stages separated by 0.5 degrees C steps between 36.0 and 38.5 degrees C. The larger gains were recorded between higher temperatures at high flows, between mid temperatures at intermediate flows, and between lower temperatures at low flows. The critical temperature for the calculated maximal gain was directly correlated to superfusion flow. The basal frequencies were consistently elevated when switching to saline equilibrated with 20% O2 and no significant differences in mean ranks were recorded between 36 and 37 degrees C, as between 38 and 39 degrees C, but frequencies at 36-37 degrees C were significantly higher than those at 38-39 degrees C. Brief rises in chemosensory discharges were evoked by injections of NaCN applied to carotid bodies superfused with saline equilibrated with 100% O2. The least effective dose was lower at 40 degrees C than at 37.5 degrees or 35.0 degrees C, but the reactivity and slope were not significantly different. It is concluded that the carotid body chemoreceptors fulfill the criteria for being considered as thermosensors, and that their frequency of discharges is thermally modulated within a range close to physiological body temperature. PMID- 8422579 TI - The formalin test in the naked mole-rat (Heterocephalus glaber): analgesic effects of morphine, nefopam and paracetamol. AB - The present experiments were initiated to study the effects of morphine, nefopam and paracetamol in the naked mole-rat, a hairless rodent that lives in subterranean colonies of up to 300, following the inability to demonstrate morphine analgesia in the hot-plate test in the rodent. The formalin test was used. Injection of 20 microliters 10% formalin produced two periods of high licking and pain behaviour, the early (0-5 min) and the late phase (15-60 min). Morphine (10 or 20 mg/kg), nefopam (10 or 20 mg/kg) and paracetamol (200 mg/kg) significantly inhibited the two phases. Paracetamol (400 mg/kg) produced significant analgesia only during the late phase. It is concluded that, unlike in the hot-plate test, it is possible to demonstrate the analgesic effects of morphine in the naked mole-rat, in the formalin test. PMID- 8422580 TI - Characterization of [3H]cytisine binding to human brain membrane preparations. AB - The binding characteristics of [3H]cytisine, a putative CNS nicotinic receptor ligand, were examined in 4 regions of the human brain. [3H]Cytisine was found to bind non-cooperatively with high affinity to a single site in tissue homogenates and to exhibit low non-specific association. The binding characteristics of this ligand were evaluated in thalamus at 4 degrees C and 24 degrees C. The association constants were found to be 0.234 and 0.308 min-1 nM-1, while the dissociation constants were 0.007 and 0.098 min-1, respectively. Saturation analysis of thalamus revealed the equilibrium Kd to be 147 pM (4 degrees C) and 245 pM (24 degrees C), values in good agreement with those determined kinetically. The Hill coefficient varied slightly between brain regions; however, the mean values in all regions examined were close to 1.0 at 0.95 +/- 0.03 (4 degrees C) and 0.91 +/- 0.04 (24 degrees C). [3H]Cytisine binding could be displaced using both nicotinic agonists and antagonists. Cytisine was the most potent displacer of [3H]cytisine binding with an Ki of 250 pM. Nicotine and acetylcholine were also potent displacers with Ki values of 1.8 and 8.1 nM, respectively. The nicotinic antagonists alpha-bungarotoxin and mecamylamine were ineffective competitors for the [3H]cytisine binding site while dihydro-beta erythroidine had an Ki value of 109 nM. Thalamus showed the highest density of cytisine binding sites of all the regions examined (48 fmol/mg protein) while the hippocampus, cingulate gyrus and the cortex showed Bmax values of 18.9, 19.3 and 8.8 fmol/mg protein, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422581 TI - Distribution of parvalbumin immunoreactivity in the vertebrate retina. AB - Parvalbumin, a calcium-binding protein thought to buffer intracellular calcium, is expressed in selected neuronal and non-neuronal cell populations. We used a well-characterized antibody directed against parvalbumin to investigate the distribution of parvalbumin in the retina of twelve vertebrate species to evaluate patterns of cellular expression for recurrent functional features. Parvalbumin immunoreactivity was displayed by subpopulations of ganglion, amacrine, bipolar and horizontal cells in different species-specific combinations. In the pigeon retina, subpopulations of amacrine, ganglion and bipolar cells were immunoreactive for parvalbumin. Parvalbumin immunoreactive bipolar cells in this species were mostly confined to the temporal dorsal region of the retina. In the owl, no immunoreactive amacrine cells were found, but many bipolar cells displayed parvalbumin immunoreactivity. In the teleost retina, amacrine and ganglion cells were found to be immunoreactive for parvalbumin. A high degree of species-specific variation was encountered in the mammalian retina. The most consistent finding within this class was that subpopulations of parvalbumin-immunoreactive amacrine cells were consistently observed in every species. In the rabbit, horizontal and ganglion cells displaying parvalbumin immunoreactivity were also seen. In rodents (hamster, ground squirrel), parvalbumin immunoreactivity was displayed by subpopulations of amacrine cells and, in the squirrel, by some ganglion cells as well. In the cat and in the baboon retina, parvalbumin immunoreactivity was found in horizontal cells, ganglion cells and a subpopulation of amacrine cells. The distribution of parvalbumin immunoreactive neurons in the vertebrate retinae studied showed no systematic correlation with phylogenetic proximity. The expression of parvalbumin within the systems of retinal neurons may therefore reflect the functional needs of different visual behaviors. PMID- 8422583 TI - Gliosis in human brain: relationship to size but not other properties of astrocytes. AB - Gliosis is the most frequent and therefore important neurocellular reaction to brain insult occurring in diseases ranging from AIDS to infarction. Neuropathological diagnosis of gliosis is based on morphological changes of brain glial cells. Changes commonly agreed to reflect gliosis are qualitative increases in size, number and glial fibrillary acidic protein (GFAP) immunoreactivity of astrocytes. These parameters were morphometrically quantified in brain tissues of 22 individuals who died with 7 diseases and statistically compared to the extent of gliosis independently determined by 3 qualified observers. The data indicate that the extent of gliosis correlated with the increase in size of astrocytes in white matter (p = 0.67) and this relationship was statistically significant (P = 0.0006). In contrast, the extent of gliosis was not correlated with the density of astrocytes nor the intensity of GFAP staining. PMID- 8422582 TI - Differential response of striatal dopamine and muscarinic cholinergic receptor subtypes to the loss of dopamine. III. Results in Parkinson's disease cases. AB - The distribution and number of DA uptake sites, DA receptors (D1 and D2) and cholinergic muscarinic receptors (M1 and M2) were examined by autoradiography in the striatal complex of Parkinson's and age-matched control cases. The greatest loss of DA uptake sites occurred in the dorsolateral striatum which was the only region showing an increase in D2 receptors. The number of M2 receptors was reduced in the dorsolateral striatum and M1 receptors were reduced in most regions of the striatum. The anatomical pattern of changes in DA uptake sites, D2 receptors and M2 receptors suggests a coordinated change in the dopaminergic cholinergic interneuron synapse in Parkinson's disease. PMID- 8422584 TI - Bulbospinal respiratory neurons are a source of double synapses onto phrenic motoneurons following cervical spinal cord hemisection in adult rats. AB - The purpose of this study was to determine if the medullary neurons that provide the primary excitatory drive to phrenic motoneurons (i.e., rostral ventral respiratory group, rVRG) are a source of double synapse formation in the phrenic nucleus after spinal cord hemisection. The axons of rVRG neurons either ipsilateral or contralateral to the hemisection were labeled by injection of a mixture of HRP and WGA-HRP into the rostral ventral respiratory group. Phrenic motoneurons ipsilateral and caudal to the hemisection were labeled by the retrograde transport of HRP. The ultrastructural results indicated that after hemisection, rVRG neurons from both sides of the medulla formed labelled double synapses in the phrenic nucleus. PMID- 8422585 TI - Electrophysiology and burst-firing of rat subicular pyramidal neurons in vitro: a comparison with area CA1. AB - Intracellular recordings were made from subicular and CA1 neurons in slices of the ventral hippocampal and parahippocampal region of the rat. All of the subicular cells that were stained by intracellular injection of biocytin were pyramidal in form. Although most electrophysiological properties were similar in the two areas, in response to depolarising current injection, the majority of subicular cells displayed a distinctive pattern of burst-firing which was rarely seen in CA1 cells. Burst-firing was voltage sensitive but was not abolished by blocking excitatory synaptic transmission, suggesting that it is an intrinsic membrane property of subicular pyramidal cells. PMID- 8422586 TI - Medial region of the amygdala: involvement in adrenal-steroid-induced salt appetite. AB - We report the following observations: first that cell body damage induced by ibotenic acid within the medial region of the amygdala resulted in the abolition of corticosterone-potentiated, aldosterone-induced salt appetite. Sodium depletion-induced salt appetite was not impaired by the lesion, nor was angiotensin-dependent salt appetite via captopril treatment. These results provide evidence that the medial region of the amygdala has a major role in the natriorexigenic effects of the adrenal steroids. PMID- 8422587 TI - Prolonged inhibition of the flexor reflex by probing the cervix uteri in the cat. AB - In decerebrate or spinal cats, sustained mechanical stimulation of the cervix uteri inhibited the flexor reflex elicited by electrical stimulation of the foot pad during the probing period (160 s). After probing, 3-15 min were required for reflex recovery. No additional inhibition was produced if probing was repeated before recovery, but instead the reflex was facilitated. When probing was applied 5-10 min after reflex recovery the reflex was again abolished. The recovery, however, occurred earlier and was followed by facilitation. Probing the cervix with single mechanical pulses inhibited transiently (140-200 ms) the short latency reflex components, but the components with longer latencies are unaffected or facilitated. Distension of the vaginal wall with a balloon also inhibited the flexor reflex, but a transient, mild facilitation appeared several seconds after the distension. In general, whenever the inhibition decreases, the facilitation predominates. Our findings suggest that cervical probing or vaginal distension triggers both a long-lasting inhibition and a concomitant facilitation in different intraspinal flexor reflex pathways. PMID- 8422588 TI - Sustained activation of the triceps surae muscles produced by mechanical stimulation of the genital tract of the female cat. AB - In decerebrate cats, controlled mechanical stimulation of the perivulvar skin, the vaginal wall or the cervix uteri induced visible hind limb extension. Pressing on the cervix uteri produced the greater response. To quantify these responses, the EMG activity and the tension developed by the normally inserted triceps surae muscles were recorded. The activity induced in these muscles by stimulation of the genital canal outlasted the stimulus by many seconds or a few minutes. These effects disappeared after spinalization at the T12 level. We propose that stimulation of the vaginal canal in the female cat may induce bistability of triceps surae motoneurones. PMID- 8422589 TI - CGRP-like immunoreactivity in A11 dopamine neurons projecting to the spinal cord and a note on CGRP-CCK cross-reactivity. AB - Using the indirect immunofluorescence technique and double labelling procedures combined with retrograde tracing it could be demonstrated that the A11 dopamine cell group, located at the border between the diencephalon and mesencephalon of the rat brain and some of which project to the spinal cord, contains calcitonin gene-related peptide (CGRP)-like immunoreactivity. Thus, another catecholamine group in the rat brain has been shown to have a coexisting peptide. One of the CGRP antisera used in the present study also stained cholecystokinin (CCK) containing neurons in various brain areas. Absorption and displacement experiments using immunohistochemistry and radioimmunoassay showed that this cross-reactivity was confined to the C-terminal portion of the peptide molecule. Therefore, the present results suggest that CGRP antisera used for immunohistochemistry and radioimmunoassay should be tested for possible cross reactivity with CCK. PMID- 8422590 TI - Trophic effects of interleukin-4, -7 and -8 on hippocampal neuronal cultures: potential involvement of glial-derived factors. AB - The main purpose of the present study was to determine whether specific lymphokines may be neurotrophic, by testing their effects on the survival of hippocampal neuronal cultures. Previous studies have shown that a variety of interleukins may be neurotrophic or neurotoxic, depending upon the culture conditions, as well as the concentration and time of exposure to the interleukins. The present results indicate that interleukins-4, -5, -7 and -8 significantly enhance neuronal survival of hippocampal cultures. These effects were concentration-dependent and reached maximal levels with concentrations of the lymphokines ranging from 500 to 1,000 ng/ml. With increased exposure to the lymphokines, the increase in neuronal survival compared to control untreated cultures persisted. Moreover, with IL-7, and particularly IL-8, this increased survival was more pronounced in the longer-term cultures. Thus, in the 7-day-old cultures, the magnitude of the increase in survival in the IL-8-treated cultures ranged from 93 to 123% compared to 56-68% in the 3-day-old cultures. In contrast, other lymphokines tested, interleukin-3 and -6, did not affect the survival of 1 day-old cultures and caused significant reductions in the longer-term cultures. Although the mechanism(s) of the neurotrophic effects of interleukins-4, -7 and 8 are not clear, an indirect effect mediated by proliferating glia in the treated cultures may be possible. Clearly, exposure to interleukins-4, -7 and -8 resulted in a marked increase in the number of astroglia and microglia compared to the control cultures, an effect that was amplified with increased time in vitro.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422591 TI - Pyramidal cells in piriform cortex receive a convergence of inputs from monoamine activated GABAergic interneurons. AB - Previously, serotonin (5-HT) was shown to increase inhibitory post-synaptic potentials (IPSPs) in layer II pyramidal cells, and excite a subpopulation of interneurons located on the layer II/III border of piriform cortex in rat in vitro brain slices. In the present study, the effects of norepinephrine (NE) and dopamine (DA) on these two populations of neurons were examined in brain slices using intracellular and extracellular recordings. All three monoamines increased GABAergic IPSPs in many pyramidal cells; overall, 5-HT was most effective in eliciting IPSPs (58% of cells), followed by NE (45%), then DA (24%). Commonly, pyramidal cells responded with an increase in IPSPs to more than one of the monoamines. The increase in IPSPs was found to include an increase in the frequency of IPSPs present at baseline, as well as recruitment of additional IPSPs of different amplitudes. In interneurons the effects of the monoamines paralleled that which was found for the pyramidal cells. Thus, all three monoamines increased the firing rate of many interneurons; again 5-HT was most effective (56%), followed by NE (51%), then DA (42%). In about 10% of the interneurons the monoamines inhibited cell firing. Interneurons frequently had responses to more than one of the monoamines. The excitatory amino acid (EAA) antagonist, kynurenic acid (200-400 microM), spared most 5-HT and NE responses on interneurons, suggesting that these effects were directly mediated. We conclude that IPSPs elicited by monoamines in pyramidal cells result from a convergence of inputs from populations of layer II/III interneurons that are activated by one, two or all three of the monoamines. PMID- 8422592 TI - Reversal of LTP by theta frequency stimulation. AB - Reversal of long-term potentiation (LTP) by physiological stimulation was tested in the CA1 field of hippocampal slices. In control medium, a one minute episode of 5 Hz (theta frequency) stimulation beginning 1-3 min after LTP had no effect on the degree of potentiation measured 30 min later. However, in the presence of norepinephrine (200 microM), 5 Hz stimulation reduced LTP by about 30%. Theta frequency stimulation was only effective when administered within 10 min of LTP induction and had no lasting effects on non-potentiated synapses. Stimulation at 1 Hz did not reverse LTP and stimulation at 10 Hz was no more effective than 5 Hz stimulation. LTP could be nearly completely reversed by theta frequency stimulation when potentiation was induced by milder and more naturalistic stimulation patterns. Under these conditions, LTP reversal was blocked by an antagonist of adenosine A1 receptors. These results suggest that the hippocampal theta rhythm promotes both the induction of LTP and its subsequent reversal with the latter process involving activation of adenosine receptors. Reversal of LTP may function to refine or sharpen recently encoded representations. PMID- 8422593 TI - Chemotherapy consisting of cisplatin, epirubicin, and cyclophosphamide in advanced ovarian carcinoma. The Gynecologic Oncology Group of the Comprehensive Cancer Center, Limburg, The Netherlands. AB - Between January 1985 and May 1988, 87 patients with stage III-IV ovarian carcinoma were entered in a study assessing chemotherapy consisting of cyclophosphamide, 500 mg/m2, epirubicin, 75 mg/m2, and cisplatin, 50 mg/m2, intravenously (IV) on day 1, every 4 weeks (CEP-1). The results after a median follow-up of 4 years are presented. The pathologically complete remission rate was 36%. The median survival of all patients was 26 months. For patients with stage III disease debulked to lesions < or = 1.5 cm before the initiation of chemotherapy (n = 35), the median survival was 61+ months. These data are not clearly different from the results of a previous study conducted by our group utilizing CAP-1 chemotherapy. PMID- 8422594 TI - The use of lasers in colorectal cancer. AB - The potential addition of lasers to the armamentarium of physicians treating advanced colorectal cancer is welcome because of the morbidity associated with the disease and because of the limitations of standard treatments for some of the problems related to this common disease. However, laser treatment cannot be regarded yet as a standard modality. Questions still remain regarding the efficacy of these treatments in relieving the most troublesome symptoms of tenesmus and urgency, although bleeding and mucous discharge appear to be well controlled. Additional questions remain regarding expense and the relative benefits of lasers versus electrocoagulation (much cheaper) for rectal cancers. Other concerns relate to the learning curve involved with lasers, particularly for decompression of colonic obstructions, and efficacy compared to pneumatic dilatation. PMID- 8422595 TI - Edatrexate, an antifolate with antitumor activity: a review. AB - Edatrexate (10-ethyl, 10-deaza-aminopterin; 10-EdAM) is one of a group of compounds developed by substitutions at the N10-position of 4-aminofolate. In phase I and II trials, activity has been seen against non-small-cell lung cancer, breast cancer, non-Hodgkin's lymphoma, and cancer of the head and neck. In preclinical studies, a synergistic effect has been reported when edatrexate is combined with other antineoplastic drugs, and enhanced activity has been seen in two combination-chemotherapy phase II studies in patients with non-small-cell lung cancer. In in vivo preclinical studies, edatrexate has demonstrated antitumor activity against mouse solid and ascites tumors as well as human tumor xenografts. The activity is superior to that of methotrexate and the other antifolates tested. The improved therapeutic index of edatrexate appears to be related to its increased entry into, and polyglutamylation within, tumor cells, and its relative exclusion and rapid elimination from sensitive host tissues, compared to methotrexate. Edatrexate is metabolized in the liver and then excreted mainly in the bile. In clinical trials in cancer patients, the dose limiting and most frequent toxicity is mucositis. Other side effects are generally mild and include myelosuppression, nausea, vomiting, elevations in SGOT, and macular rash. The responses seen in clinical trials along with preclinical data suggest that edatrexate may be a valuable agent in the treatment of cancer. Studies currently underway include the evaluation of edatrexate in small-cell lung cancer and edatrexate in combination with leucovorin, new vinca alkaloids, and cisplatin. PMID- 8422596 TI - Offering the option of randomized clinical trials to cancer patients who overestimate their prognoses with standard therapies. AB - We have shown that cancer patients' routine (and understandable) overestimations of their prognoses with standard therapy may inhibit their accrual to randomized clinical trials for which standard therapies are the alternative. Patients' appreciation of the rationale for a trial, and the potential benefit of trial participation, can only be enhanced if they understand their prognoses with standard therapy. However, clinical investigators may be reluctant to provide specific information that deflates patients' estimates of their prognoses. The routine withholding of information regarding the modest benefits of standard therapies may avoid patient distress, but such physician behavior is paternalistic and may deleteriously affect trial accrual. On the other hand, the routine communication of prognostic information will cause significant distress among patients and will perhaps be destabilizing to that minority of patients who would otherwise shun this information or truly cannot psychologically tolerate it. A middle ground between these extremes is the stepwise disclosure of potentially distressing information, wherein specific prognostical information is offered by physicians to patients and actually provided or communicated only after patients first understand the nature of it and then indicate their interest in receiving it. A practical disadvantage of this approach is its additional demand on physicians' time. Therefore, if impracticality is to be avoided and yet the approach fostered, clinical investigators might consider developing trial specific, written or audiovisual materials for patient education about general background information. These could be employed prior to patient-physician dialogue and so enable physicians to focus on more sensitive subjects, such as prognosis with standard therapy. PMID- 8422597 TI - An intensive sequenced adjuvant chemotherapy regimen for breast cancer. AB - Twenty-two women (17 pre- and 5 postmenopausal) with nodepositive, stage II breast carcinoma were treated with adjuvant chemotherapy consisting of 16 weeks of intensive CMFVP (cyclophosphamide, methotrexate, 5-fluorouracil, vincristine, and prednisone) administered in the original dose and schedule of the "Cooper regimen," followed by four monthly, 3-day cycles of escalating doxorubicin. Toxicity was related primarily to myelosuppression associated with the doxorubicin component of the treatment regimen. All patients recovered without sequelae. No patient developed significant cardiac toxicity. With a median follow up of 43 months (range: 20-89 months), two postmenopausal patients and one premenopausal patient have relapsed at 35, 37, and 42 months, respectively. By Kaplan-Meier survival analysis, there is an 80.5% chance of disease-free survival to 42 months. The feasibility of administering adjuvant CMFVP followed by intensive doxorubicin has been established. The pilot results warrant comparative trial with the best of current regimens. PMID- 8422598 TI - Commitment to a world without cancer. AB - The author is a layperson, health educator, and political oncologist who is, in addition to her work at the cancer center at UCLA, a long-time volunteer with the American Cancer Society. She has served the National Cancer Institute as a member of the National Cancer Advisory Board and is currently a member of its Board of Scientific Counselors, Division of Cancer Control. In these various roles she was part of the lay leadership that influenced and cooperated with the Nixon administration in its efforts to pass the National Cancer Act of 1971. She has participated continually in the implementation of the National Cancer Program and Act since 1971. This essay reviews the history of our nation's support of cancer research, commencing with the founding of the American Cancer Society. It considers the factors that led to the passage of the National Cancer Act in 1971. It then comments on the interaction between universities, the National Cancer Institute, and the Cancer Act, which has produced the "cancer business" that has come of age since the Cancer Act was passed. This commentary does not review the advances made in research and clinical practice as a result of the passage of the National Cancer Act of 1971. Others, better qualified, will do that in this journal and in other publications as well. These views are concerned with the process, the systems, and ancillary activities that have developed as a consequence of the national will to conquer cancer. They are meant to highlight several portions of the whole. The "seeds" of cancer consciousness sown early by the American Cancer Society and those so firmly planted in research and in the creation of a national network of researchers, patient care institutions, and information systems by the National Cancer Institute and the National Cancer Program Plan and Act of 1971 have reaped a harvest of independent programs and even businesses that have given substance to the hopes of those seeking to achieve a world without cancer. PMID- 8422599 TI - The impact of health care reform on reimbursement for investigational therapy. PMID- 8422600 TI - Retroviruses and cancer: models for cancer in animals and humans. PMID- 8422602 TI - Best papers on leukemia. PMID- 8422601 TI - Genetic changes in prostate carcinoma cells. PMID- 8422603 TI - Recent incidence trends for breast cancer in women and the relevance of early detection: an update. AB - The incidence of breast cancer in women has been rising dramatically in the United States since 1982, based on data collected by the Surveillance, Epidemiology, and End Results (SEER) program of the National Cancer Institute. An additional three years of incidence and mortality data are included in this update of the earlier analyses; however, the focus of this report remains on examining the steeply increasing incidence trend between 1982 and 1986. Over this period, incidence rates for in situ and localized invasive tumors increased among women age 50 and older, while rates for regional and distant tumors remained stable. The incidence of localized tumors less than 1.0, 1.0 to 1.9, and 2.0 to 2.9 cm in size increased more rapidly than that of tumors 3.0 cm and greater among women over age 50. Survival rates also improved among cases diagnosed over this time period. These descriptive data suggest that early detection may be playing a role in the recent increase in the incidence of breast cancer in women, although other factors cannot be ruled out. Conclusions regarding improved cancer control await confirmation by reduced breast cancer mortality. PMID- 8422604 TI - Defining and updating the American Cancer Society guidelines for the cancer related checkup: prostate and endometrial cancers. PMID- 8422605 TI - Questionable methods of cancer management: hydrogen peroxide and other 'hyperoxygenation' therapies. AB - "Hyperoxygenation" therapy--also called "oxymedicine," "bio-oxidative therapy," "oxidative therapy," and "oxidology"--is a method of cancer management based on the erroneous concept that cancer is caused by oxygen deficiency and can be cured by exposing cancer cells to more oxygen than they can tolerate. The most highly touted "hyperoxygenating" agents are hydrogen peroxide, germanium sesquioxide, and ozone. Although these compounds have been the subject of legitimate research, there is little or no evidence that they are effective for the treatment of any serious disease, and each has demonstrated potential for harm. Therefore, the American Cancer Society recommends that individuals with cancer not seek treatment from individuals promoting any form of hyperoxygenation therapy as an "alternative" to proven medical modalities. PMID- 8422606 TI - Current trends in breast cancer. PMID- 8422607 TI - Questionable methods of cancer management: Cancell/Entelev. PMID- 8422608 TI - Breast reconstruction. PMID- 8422610 TI - History, general overview, and protection of health care workers. AB - This article provides a background so that the intensivist can appreciate the social and cultural context in which the AIDS epidemic developed. The upheaval that AIDS has caused in the gay community is described along with some of the economic effects AIDS has had on that population and on the practice of medicine, both locally and nationally. Also reviewed are changes that have taken place in the handling of patients and biologic secretions in order to protect health care workers from contamination from the causative virus. Finally, this article discusses changes in the AIDS patient population and clinical manifestations of the disease. PMID- 8422609 TI - Cancer statistics, 1993. PMID- 8422611 TI - Outcome of intensive care for patients with AIDS. AB - Data on the outcome of intensive care for patients with AIDS have been reported since 1984. Most published statistics focus on the outcome of respiratory failure due to Pneumocystis carinii pneumonia, the most frequent reason for admission to the ICU. Survival appears to be improving in recent years, partly due to the use of adjunctive corticosteroid therapy. PMID- 8422612 TI - Ethical considerations in the treatment of AIDS patients in the intensive care unit. AB - The treatment of patients with AIDS in the ICU presents the clinician with special challenges. The admission policies of ICUs are examined, and the authors suggest ways in which ethical difficulties may be minimized. A new concept of futility is suggested, which considers both the patient's holistic needs together with his or her immediate medical prognosis. Changes in the law regarding patient choice are discussed. PMID- 8422613 TI - Advance directives for patients with human immunodeficiency virus infection. AB - A consensus has emerged in recent years that decisions about life-sustaining treatment made by informed, competent patients should be respected. Advance directives are expressions of a patient's preferences regarding life-sustaining treatments made while the patient is still competent, to be used if the patient loses competence. Patients infected with HIV have a high mortality rate and frequently lose competence as their disease progresses to AIDS. Therefore, the completion of advance directives in this patient group is vitally important. PMID- 8422614 TI - Mask continuous positive airway pressure in AIDS. AB - FMCPAP and NCPAP therapy is an effective modality for treating patients with PCP, hypoxia, and respiratory insufficiency. The therapy decreases intrapulmonary shunting and improves oxygenation. It is safe and, in some cases, can be provided outside of an intensive care unit. The major benefit of MCPAP is that it postpones (and sometimes obviates) the need for intubation and mechanical ventilation. This may provide adequate time for a trial of therapy, education, ethical discussions, and completion of personal matters by patients. It is conceivable that failure to respond to MCPAP may provide prognostic information to help guide further therapy. Further outcome studies are needed to clarify this issue. Adding MCPAP to mechanical ventilation and conventional mask oxygen therapy increases the options that practitioners can use to provide the best titrated care for their patients. PMID- 8422615 TI - Acute respiratory failure due to Pneumocystis carinii pneumonia. AB - Pneumonia caused by Pneumocystis carinii is the most frequent indication for admission of AIDS patients to intensive care units. In this article, an approach to the diagnosis and management of this condition will be presented along with prognostic information. Differential diagnosis will be discussed, and characteristic responses to current standard and alternative chemotherapeutic agents and modes of ventilatory support will be reviewed. PMID- 8422616 TI - Neurologic critical care in patients with human immunodeficiency virus 1 infection. AB - Neurologic illness related to HIV-1 infection can involve the entire neuraxis and result in a wide range of neurologic syndromes. This article reviews intracerebral mass lesions, meningitis, myelopathy, peripheral neuropathy, and occupational exposure to HIV-1. PMID- 8422617 TI - Serious gastrointestinal disorders associated with human immunodeficiency virus infection. AB - Disorders of the gastrointestinal tract are a frequent complication of AIDS. Although the majority of these processes are not life threatening, some diseases will require ICU monitoring. With the increasing longevity of the HIV-infected patient, many more patients may be admitted to the ICU for a variety of other complications where gastrointestinal disorders may be manifested. This article focuses on the differential diagnosis for the more common gastrointestinal complications of HIV infection as well as a rational approach for the ICU setting. PMID- 8422618 TI - Predicting survival in AIDS patients with respiratory failure. Application of the APACHE II scoring system. AB - This article describes the APACHE II classification system as a measure of severity of illness applied to AIDS patients with respiratory insufficiency. Among 82 patients, observed mortality in patients with high APACHE II scores (greater than 30) and those with low scores (less than 18) was significantly higher than predicted. There was variable correlation between predicted and observed mortality in the other APACHE II score ranges. The usefulness of the APACHE II scoring system is reviewed as limited and inaccurate in predicting survival rates in AIDS patients with respiratory failure. PMID- 8422619 TI - Swallowing function in patients with esophageal cancer treated with concurrent radiation and chemotherapy. AB - BACKGROUND: Major goals of concurrent radiation and chemotherapy in the treatment of esophageal cancer are the early restoration and long-term maintenance of swallowing function. The purpose of this study was to determine the impact of concurrent radiation and chemotherapy on swallowing function. METHODS: Between September 1980 and September 1990, 120 patients with esophageal cancer were treated at the Fox Chase Cancer Center on the basis of one of three prospective nonrandomized protocols using concurrent chemotherapy and radiation. Swallowing function was retrospectively assessed in these patients by use of a swallowing function scoring system. In addition, patients who had long-term control of their esophageal cancer underwent a more detailed analysis of swallowing function. RESULTS: Initial improvement in dysphagia occurred in 88% of the 102 assessable patients, with a median time to improvement of 2 weeks. There was no difference in overall percentage of initial improvement for patients with adenocarcinoma versus squamous cell carcinoma. Patients with distal tumors, however, showed both earlier and higher frequency of initial improvement than did patients with tumors in the upper two-thirds of the thoracic esophagus (95% versus 79%). Local relapse free survival of definitively treated patients at 3 years was 60% and was significantly better for patients with Stage I (76%) versus Stage II cancers (55%) (P < 0.05). All 25 patients treated with curative intent who survived for more than 1 year without evidence of disease were able to eat soft or solid foods and had a benign stricture rate of only 12%. Even in patients with advanced disease who were treated with palliative intent, 91% had an initial improvement in swallowing function and 67% had improvement in swallowing function that lasted until death. CONCLUSIONS: High-dose concurrent radiation and chemotherapy provides rapid improvement in dysphagia, and this improvement results in normal or near-normal swallowing function of long duration. PMID- 8422620 TI - The association of Helicobacter pylori with gastric cancer and preneoplastic gastric lesions in Chiapas, Mexico. AB - BACKGROUND: Helicobacter pylori recently was identified as a risk factor for gastric cancer. Its association with preneoplastic conditions of the stomach, however, is undocumented. METHODS: Gastric biopsy specimens from 245 symptomatic patients were examined for neoplastic and preneoplastic lesions and for gastric H. pylori infection. The sera of 183 subjects were tested by enzyme-linked immunosorbent assay (ELISA) for anti-H. pylori immunoglobulin G. RESULTS: Histologic H. pylori infection, usually accompanied by acute and chronic gastritis, was found in 85.7% of patients. There was a strong association between H. pylori in the tissue and atrophy (relative risk, 15.0; 95% confidence interval, 4.2-56.6), intestinal metaplasia (relative risk, 5.7; 95% confidence interval, 1.9-16.8), and dysplasia or cancer (relative risk, 4.0; 95% confidence interval, 1.1-14.8). The ELISA was 93.2% sensitive and 57.1% specific for histologic infection with a positive predictive value of 96.1%. The overall seroprevalence rate was 86.1%, with no significant difference in rates between patients with cancer precursors and those with normal stomachs. CONCLUSIONS: In this high-risk population, precursor lesions for adenocarcinoma were associated universally with H. pylori infection. PMID- 8422621 TI - Diagnostic factors in gastric cardia cancer invading the esophagus. AB - BACKGROUND: The prognosis of patients with tumors in the upper one-third of the stomach, particularly those with esophageal invasion, is poor. METHODS: This study involved 168 patients with advanced cancer in the upper third of the stomach whose lesion had had invaded the esophagus. Clinical and pathologic studies were performed with respect to diagnostic factors and histologic differentiation of the lesion. RESULTS: Eighty-three patients (49.4%) had differentiated gastric cancer, and 85 (50.6%) had undifferentiated cancer. The survival time was shorter for patients with undifferentiated cancer than for those with differentiated cancer (P < 0.01). Multivariate analysis showed operative curability, liver metastasis, peritoneal dissemination, lymph node metastasis, serosal invasion, and tumor size to be independent prognostic factors. In patients with undifferentiated cancer, tumors were larger, serosal invasion was prominent, lymph node metastasis was present in 85.9% of cases, 64.7% of metastases were noncuratively resected, and survival time was less favorable. CONCLUSIONS: Our analysis shows that a group of patients at increased risk for tumor advancement will benefit from more aggressive therapy. PMID- 8422622 TI - Cytogenetic aberrations in colorectal adenocarcinomas and their correlation with clinicopathologic features. AB - BACKGROUND: Little is known about the karyotypes of colorectal carcinomas and, in particular, about how the cytogenetic findings correlate with clinicopathologic features. METHODS: Short-term cultures from 49 colorectal adenocarcinomas were analyzed cytogenetically. The karyotypes were correlated with grade, stage, lymphocytic infiltration, and site (using the chi-square test), with patient age and tumor size (using the Student t test), and with survival (using the log-rank or Mantel-Haenszel test). RESULTS: Normal karyotypes were detected in 17, simple numeric changes in 22, and multiple structural and numeric abnormalities in 10. The most common numeric aberrations were +7, -Y, -18, and -22. The most common structural rearrangements were, in decreasing order of frequency, of chromosomes 1 (eight samples, leading to loss of 1p material in five), 3, 11, 17, 6, 8, 13, and 20. Marked or moderate lymphocytic infiltration was seen significantly less often (P < 0.05) in tumors with complex chromosomal abnormalities than in those with simple anomalies or normal karyotypes. The subset of patients who had tumors with multiple chromosomal abnormalities had a significantly shorter survival time (P < 0.025) than those who had lesions with simple changes or normal karyotypes. CONCLUSIONS: Loss of 1p material is the most consistent chromosomal change in colorectal carcinomas but probably represents a progressional rather than a primary event. Structural changes of chromosomes 3 and 11 seem to be more common in tumors located in the distal part of the large intestine. The significantly shorter survival time of patients with complex aberrations indicates that the karyotype could be used as a prognostic parameter in patients with colorectal cancer. PMID- 8422623 TI - Liver metastases with 10 human colon carcinoma cell lines in nude mice and association with carcinoembryonic antigen production. AB - BACKGROUND: The secretion of carcinoembryonic antigen (CEA) by many colorectal tumors is associated with a worse prognosis and a greater likelihood of metastases. The exact biologic function of CEA is not known. In the literature, it has been postulated CEA may be a tumorigenicity-enhancing factor. METHODS: Ten different human colonic adenocarcinoma cell lines (RW-7213, RW-2982, LS174T, SW1116, RW-5928, DLD-2, SW-48, DLD-1, SW-480, and HCT-8) with a wide range of CEA production (from undetectable to 5200 ng/ml in culture medium) were injected into the spleens of groups of nude mice as a model for experimental hepatic metastasis. RESULTS: There was a wide range in local tumorigenicity in the spleen (from 0-90%) and in liver metastases (from 0-70%). The capacity to grow in both liver and spleen was associated with CEA production. The four cell lines that secreted the highest amounts of CEA produced the highest tumorigenicity in the spleen (67-90%) with frequent liver colonization (25-70%). The two cell lines that secreted no detectable CEA produced neither splenic tumors nor hepatic colonies. Low-level CEA production was associated with intermediate and more variable tumorigenicity. CONCLUSIONS: There was an association between CEA secretion and the ability of 10 different colorectal cell lines to grow in nude mouse spleen and liver models. PMID- 8422624 TI - Evaluation of combination chemotherapy and phase II agents in pancreatic adenocarcinoma. A Southwest Oncology Group study. AB - BACKGROUND: Pancreatic carcinoma responds poorly to conventional chemotherapy. To identify potentially useful agents, a sequence of clinical trials were done. METHODS: A series of Phase II randomized trials were done by the Southwest Oncology Group in which patients with metastatic or advanced pancreatic cancer were randomized to receive single agents (methylglyoxal-bis-guanylhydrazone [MGBG], dihydroxyanthracenedione [DHAD], and aziridinylbenzoquinone [AZQ]) or a combined regimen of 5-fluorouracil, doxorubicin, mitomycin C, and streptozotocin (FAM-S). Toxicity, response, and survival were determined. RESULTS: Seventy-one patients received FAM-S and 82, the Phase II single agents. Response rates (95% confidence intervals) for the various treatments were: FAM-S, 11% (0%, 21%); MGBG, 6% (0.8%, 21%); DHAD, 0% (0%, 12%); and AZQ, 0% (0%, 16%). The median survival times were: FAM-S Group, 4.8 months and Phase II agent Group, 3.4 months. CONCLUSIONS: The FAM-S regimen and the Phase II agents tested did not have substantial antitumor activity in pancreatic cancer. The use of new agents as initial therapy is reasonable. PMID- 8422625 TI - Antibody affinity disturbance in the immunoglobulin immune response to mucosal antigenic stimulation in lung cancer. The Betalactoglobulin Model. AB - BACKGROUND: This study is a continuation of a recent study, in which a defect in the immunoglobulin G (IgG) response to some natural antigens (bovine betalactoglobulin [BLG] from cow's milk and antigen p1 from the house dust mite Dermatophagoides pteronyssinus), usually presented at the mucosal level, was documented in lung cancer patients. The present study further characterizes this difference in terms of antibody relative functional affinity in the BLG model. METHODS: Relative functional affinity was evaluated by solid-phase enzyme-linked immunosorbent assay in terms of the relationship between specific IgG retention, assessed with peroxidase-labeled protein A, and serial dilutions of IgG fractions isolated from 24 sera from lung cancer patients and 24 sera from healthy control subjects matched for their anti-BLG IgG antibody titers. The procedure was performed in the presence and absence of low concentrations of diethylamine, which was expected to prevent low-affinity antigen-antibody binding without affecting the binding of high-affinity antibodies. Anti-BLG IgG antibody affinity also was evaluated in 25 patients with early-stage lung cancer, before and after (5 +/- 1 week) complete surgical excision of the tumor. RESULTS: Results, expressed as the slope of the binding curves and their leftward shift induced by diethylamine, showed different antibody populations between the two groups. Control sera showed a heterogeneous population of anti-BLG IgG antibodies, including antibodies of higher (steeper slope) and lower (more gradual slope) functional affinity. Cancer sera exhibited a less heterogeneous population of anti-BLG IgG antibodies, mostly with lower functional affinity. No change was observed in anti-BLG IgG antibody affinity in the 25 lung cancer patients tested 5 +/- 1 week after complete surgical excision of the tumor. CONCLUSIONS: These results document a persistent qualitative immunologic disturbance in patients with lung cancer, regardless of the type and extent of tumor. The potential relationship between this observation and the development of lung cancer, however, is presently unknown. PMID- 8422626 TI - Osteosarcoma. Low-grade intraosseous-type osteosarcoma, histologically resembling parosteal osteosarcoma, fibrous dysplasia, and desmoplastic fibroma. AB - BACKGROUND: Low-grade intraosseous osteosarcoma is a rare variety of osteosarcoma and it is difficult to recognize. METHODS: A series of 10 patients with low-grade intraosseous osteosarcoma is reported. These patients were identified from the Istituto Rizzoli files, which includes approximately 1000 cases of osteosarcoma. Clinical data and radiographic and histologic features were studied. RESULTS: The radiographic appearance confirmed malignancy in five patients and suggested it in two. A benign lesion was diagnosed in three patients. Three lesions resembled parosteal osteosarcoma, two appeared similar to fibrous dysplasia, and two had features of desmoplastic fibroma. A mixed histologic pattern was found in three other tumors. Recurrence after intralesional excision in all patients indicated the aggressive nature of this lesion. The development of metastases in two patients and progression in the grade of malignancy in one of these highlighted the malignant nature of the tumor. CONCLUSIONS: The correct diagnosis would permit adequate treatment with wide surgical margins. PMID- 8422627 TI - Aggressive osteoblastoma of the calcaneus. AB - Aggressive osteoblastoma of the left calcaneus in 29-year-old Japanese woman is reported. Her initial symptom was heel pain while walking. This was a primary calcaneal tumor, initially diagnosed as a benign osteoblastoma. After a 5-year follow-up (from the initial curettage), there was local recurrence. The histologic findings of aggressive osteoblastoma were confirmed after right lower leg amputation. The recurrent tumor was mildly aggressive to the talocalcanean joint and the retrocalcaneal area, without distant metastasis. The characteristics of the primary and recurrent tumors were examined by the radiologic, histologic, and electron microscopic procedures. Although there are questions about aggressive osteoblastoma, the authors believe that there are osteoblastic tumors of borderline malignancy between benign osteoblastoma and low grade osteosarcoma. The current case was an example compatible with an aggressive osteoblastoma with the proposed name of Dorfman classification Group 4. PMID- 8422628 TI - Acalculous cholecystitis in bone marrow transplant patients. AB - BACKGROUND: Acalculous cholecystitis (ACC) is an uncommon disorder of the biliary tract, accounting for approximately 6% of acute cholecystitis cases. In this study, cholecystitis was seen in 8 of 770 bone marrow transplant recipients, with ACC occurring in five (63%). METHODS: Records of 592 allogenic and 150 autologous BMT patients were reviewed for risk factors associated with ACC. RESULTS: Only the number of blood transfusions administered and the use of total parenteral nutrition were associated with ACC development. ACC occurred in 4 of 42 (9%) allogeneic recipients who required exchange transfusion for ABO incompatibility. ACC developed in one autogolous recipient alongside venoocclusive disease of the liver. There was no association between ACC development and preparative regimen, cyclosporine usage, graft versus host disease, or cytomegalovirus infection. CONCLUSIONS: ACC occurs more frequently in patients after ABO incompatible BMT requiring exchange transfusion than in other transplant recipients. PMID- 8422629 TI - Heterogeneous immunoglobulin gene rearrangement in a B-chronic lymphocytic leukemia progressing into non-Hodgkin lymphoma (Richter syndrome). AB - BACKGROUND: The relationship between chronic lymphocytic leukemia (CLL) and supervening non-Hodgkin lymphoma is debated, as is whether a particular genomic pattern is related to the emergence of the terminal lymphoma. To investigate these features, the molecular organization of the immunoglobulin (Ig) gene region in a case during both the B-CLL and Richter transformation phase was studied. METHODS: B-CLL and non-Hodgkin lymphoma cells were processed for Southern blot analysis of Ig heavy- and light-chain gene configuration. RESULTS: Molecular studies of B-CLL cells revealed the presence of a single Ig heavy-chain rearrangement with both kappa and lambda light-chain rearranged genes, which was consistent with the occurrence of multiple mutational events during the development of the B-CLL clone. Molecular analysis of the lymphoma DNA showed new Ig heavy- and kappa light-chain rearrangements in addition to the original ones related to the CLL phase, indicating that the lymphoma tissue consisted of two genotypically distinct populations of cells. CONCLUSIONS: On the basis of the overall molecular configuration, this heterogeneous pattern of Ig gene rearrangement was interpreted as an inherent genetic instability of the CLL clone, in which multiple mutational events allowed a selective pressure toward more aggressive subclones, resulting in the emergence of the terminal lymphoma. PMID- 8422630 TI - Effects of high-dose chemotherapy with syngeneic bone marrow transplantation in experimental brain tumor in rats. AB - BACKGROUND: The clinical dose of 3-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-1-(2 chloroethyl)-1-nitro- sourea hydrochloride (ACNU) is limited by the bone marrow suppression it produces. The authors investigated whether bone marrow transplantation (BMT) associated with a high dose of ACNU could alleviate marrow toxicity and achieve greater antitumor effects in Fischer rats. METHODS AND RESULTS: High doses of ACNU (20, 25, and 30 mg/kg) were administered on day 1, followed by injection of syngeneic bone marrow cells on day 3. Patterns of leukocyte recovery were compared in BMT and non-BMT groups. There was a significant benefit in leukocyte recovery in the BMT groups. A 9L glioma that was sensitive to ACNU was transplanted into the brain tissue of rats. Seven days later, the animals were divided into three groups based on the dose of ACNU (2, 10, and 25 mg/kg). The median survival time increased with larger ACNU doses. CONCLUSIONS: This suggested that high-dose ACNU chemotherapy with BMT could achieve greater antitumor effects and less bone marrow suppression than high-dose ACNU alone. PMID- 8422631 TI - Analysis of the cost-effectiveness ratio of the health campaign for the early diagnosis of cutaneous melanoma in Trentino, Italy. AB - BACKGROUND: The cost-effectiveness ratio of a health campaign for the early diagnosis of cutaneous melanoma (CM), currently the only means to reduce mortality from this melanoma, was calculated for the Trentino region in Italy. METHODS: Health campaigns in Trentino were carried out, and comparisons were made with neighboring regions where there had been no campaigns. To do this, the trend of the mortality rates calculated according to sex and age using the direct standardized method and data provided by ISTAT (the Central Italian Institute of Statistics) was considered. RESULTS: It was determined that 22.3 lives were saved by early diagnosis during the period 1977-1985, resulting in a savings of $494,636 to the Italian National Health Service in avoided treatments. The total cost to the Health Service of the campaign was $70,800. The cost per year of lives saved was calculated to be $400. CONCLUSIONS: The outcome of this campaign was a recommendation for the use of such programs in other countries and regions. PMID- 8422632 TI - High levels of DNA index heterogeneity in advanced breast carcinomas. Evidence for DNA ploidy differences between lymphatic and hematogenous metastases. AB - BACKGROUND: The aim of this study was to investigate DNA ploidy status and DNA index heterogeneity of lymphatic and hematogenous metastases of advanced breast carcinomas and the relations among the various tumor sites. METHODS: DNA ploidy status was analyzed by flow cytometry on frozen and paraffin-embedded tissue blocks taken from primary and metastatic tumor sites in 18 patients with advanced breast cancer. RESULTS: Presumably because of the extensive sampling, high percentages of DNA aneuploidy, DNA multiploidy, and DNA index heterogeneity were found in primary breast carcinomas as well as in lymph node and distant metastases. DNA aneuploid tumor stemlines were frequently accompanied by DNA diploid tumor stemlines. Most of the DNA tumor stemlines found in the primary tumors recurred in lymph node (55%) and distant (59%) metastases, even after 17 years of relapse-free survival. DNA tumor stemlines found in distant metastases, however, often differed from those in lymph node metastases (61%). CONCLUSIONS: A marked DNA index heterogeneity can be found in primary and metastatic tumor sites when appropriate sampling is applied. There were no DNA ploidy subclasses, notably absent in either type of metastasis, indicating similar metastatic capacities of both DNA aneuploid and DNA diploid tumor stemlines in advanced breast carcinomas. The difference in DNA ploidy status between lymphatic and hematogenous metastases suggest that these metastases can be generated independently. PMID- 8422633 TI - Feasibility of dose-intensive continuous 5-fluorouracil, doxorubicin, and cyclophosphamide as adjuvant therapy for breast cancer. AB - BACKGROUND: Several studies support the belief that the efficacy of adjuvant chemotherapy in breast cancer is related to the dose intensity of the chemotherapy used (expressed in milligrams per square meter per week). Retrospective analyses indicate that the actual delivered dose intensity may correlate more strongly with efficacy than the intended dose delivery. METHODS: A doxorubicin-based adjuvant regimen was studied for breast cancer. It was modeled on the Southwest Oncology Group's regimen of cyclophosphamide, methotrexate, and 5-fluorouracil in that daily oral cyclophosphamide and weekly intravenous 5 fluorouracil were used, but weekly doxorubicin was substituted for methotrexate and administered in both the adjuvant and neoadjuvant setting to 29 patients. RESULTS: The actual dose delivery was 1.21-1.24-fold that calculated for the delivered dose in the two other adjuvant regimens using the same three drugs (5 fluorouracil, doxorubicin, and cyclophosphamide) for which this could be determined. This regimen was tolerated well. Toxicity was minimal and consisted largely of expected neutropenia. CONCLUSIONS: Whether improved dose intensity can be increased further with the use of growth factors or actually confers improved outcomes awaits the results of larger future trials. PMID- 8422634 TI - Gene structure and expression analysis of the epidermal growth factor receptor, transforming growth factor-alpha, myc, jun, and metallothionein in human ovarian carcinomas. Classification of malignant phenotypes. AB - This study reports the structure and expression rates of genes of the transforming growth factor-alpha (TGF-alpha) signal transduction pathway (TGF alpha, epidermal growth factor receptor [EGF-R], jun, myc, and metallothionein [MT]) in 47 specimens of ovarian cancer and 21 nonmalignant tissues. The objective was to establish a direct correlation between the genetic activities and the malignant phenotype of the ovarian cancer. The Southern blot technique identified four samples with myc amplification and two with rearranged EGF-R genes. By using the S1 nuclease assay, the analysis of myc transcription showed a similar use of both promotors. Although the size of the investigated transcripts was unaltered, significant differences in the transcription rates were noticed in malignant tissue probes (using northern blot analysis and RNAase protection assay). The following results of messenger RNA analysis in ovarian cancer were observed: EGF-R, negative in 25%, low in 65%, and strongly positive in 10%; TGF alpha, negative in 34%, low in 36%, and strongly positive in 30%; myc, negative in 8%, low in 64%, and strongly positive in 28%; jun, negative in 4%, low in 58%, and strong in 38%; and MT, low in 80% and strongly positive in 20%. In most nonmalignant tissues studied, no or only a low expression of TGF-alpha, EGF-R, and myc. was found. A comparison of these messenger RNA results with the clinical data from tumors showed four different subgroups of ovarian carcinomas. The results of chemotherapy were known in 32 cases. Tumors with negative or low expression rates of all investigated genes did not respond to chemotherapy; 13 of 18 tumors with high expression rates did respond. Additional signal transduction chains distinct from the TGF-alpha pathway, however, are likely to influence both the expression and activity of transcription factors and MT. PMID- 8422635 TI - The management of ovarian carcinoma is improved by the use of cancer-associated serum antigen and CA 125 assays. AB - BACKGROUND: The new tumor-associated mucin assay, cancer-associated serum antigen (CASA), was assessed with the CA 125 assay for use in the management of patients with epithelial ovarian cancer. METHODS: CASA and CA 125 were assessed retrospectively for use in (1) monitoring 28 patients with Stage 3 or 4 ovarian carcinoma during therapy, (2) predicting the outcome of 41 second-look laparotomies (SLL), and (3) predicting the survival outcome by measuring these levels after surgery but before chemotherapy in 65 patients with Stage 3 disease. RESULTS: Of 20 patients with recurrence after an initial response, the presence of CASA levels detected recurrence in 65% before clinical detection; CA 125, 50%; and the combination of CASA and CA 125, 80%. Six patients whose disease was in long-term remission did not have elevations of either marker. When used to predict the results of SLL, the positive predictive values of CASA and CA 125 were 77% and 100%, respectively. The negative predictive values for CASA and CA 125 were 71% and 66%, respectively. CASA detected 50% of positive SLL where microscopic disease only was found; the CA 125 test did not. Multivariate analysis of survival rates using levels of CASA and CA 125, age, residual disease, tumor type and grade, or the presence or absence of cisplatin in the chemotherapeutic regimen found that postoperative CASA levels ranked above all prognostic factors except age. CASA levels may be more accurate than surgical reporting of residual disease or they may define a subset of patients with biologically more aggressive ovarian carcinoma. CONCLUSIONS: The CASA test is sensitive to ovarian carcinoma, and both CASA and CA 125 are more useful when used in conjunction. PMID- 8422636 TI - Late results in patients treated with pi-mesons for bladder cancer. AB - BACKGROUND: The treatment of bladder cancer with 60 converging pion beams was expected to have certain dose-distribution advantages with possibly fewer side effects than other high linear-energy-transfer (LET) radiation therapies, such as neutrons. RESULTS: Early results were promising: 20 of 24 (83%) evaluable patients with sessile invasive bladder carcinomas had clinically complete responses. However, only 3 of 10 (30%) evaluable patients with superficial bladder tumors had clinically complete responses. This article reports the long term follow-up (6-8 years) of these patients with emphasis on the late side effects of pion radiation therapy. Thirty-eight of the 41 (93%) patients treated died after a median survival time of 17 months (range, 4-98 months). Seventeen (45%) died of metastatic disease (in two instances, this was combined with a local recurrence) 5-27 months after radiation therapy. Four (10%) died of locally progressive disease, and eight (21%) died of late side effects of radiation therapy 9-98 months after treatment. All these patients were treated with more than 33 pion Gy and had generally a symptom-free interval of 9-18 months. The observed side effects were severe, consisting of chronic inflammation and vascular damage in the pelvic region often followed by ulceration, fistulas, and perforations throughout the intestines. In 11 patients, cystectomy and urinary diversion was necessary because of excessive fibrosis and bladder shrinkage. In eight patients, a colostomy was required for stenotic inflammatory disease, necrosis, and perforations of the intestines. The remaining nine patients (24%) died of causes unrelated to the primary disease 4-60 months after radiation therapy. CONCLUSIONS: The results of the first Phase I/II trial using the Swiss piotron showed a high complete response rate in patients with sessile bladder cancers but also a high incidence of local recurrences and severe, in some instances lethal side effects. Although it is expected that these results will be the basis for future improvements, particularly regarding dosing and fractionation, this experience emphasizes the need for a sufficiently long observation period before reaching conclusions about any high LET treatment, such as neutron, pion, or heavy ion radiation therapy. PMID- 8422637 TI - Primary malignant melanoma of the female urethra. AB - BACKGROUND: Malignant melanoma is one of the rarest tumors of the female urethra. The prognosis of urethral melanoma is poor. METHODS: A 59-year-old woman underwent total urethrectomy, bilateral inguinal lymph node dissection, and vesicostomy for Stage A primary malignant melanoma of the urethra. She received adjuvant therapy consisting of dacarbazine, vincristine, cyclophosphamide, and alpha-interferon. Two years later, repair of the parastomal hernia and interposition of an isoperistaltically ileum intussusception between the bladder and the abdominal wall were done. RESULTS: Complete continence was achieved, and the patient catheterizes herself every 3-5 hours during the day and once at night without difficulty. She has survived 5 years without any evidence of tumor recurrence. CONCLUSIONS: There have been only six case reports of women with primary malignant melanoma of the urethra surviving more than 5 years, including this case. Total urethrectomy with bilateral inguinal lymph node dissection should be done as the initial form of the treatment in patients with this disease who have no evidence of distant metastasis. The postoperative adjuvant therapy, consisting of alpha-interferon and chemotherapy, should be administered immediately; therefore, vesicostomy was done because it is a simple procedure. However, this type of urinary diversion requires an external appliance. Clinicians should consider the patient's quality of life, prognosis, and general condition and attempt to solve the problems of patients 2-3 years after the operation. PMID- 8422638 TI - Regional lymph node involvement and its significance in the development of distant metastases in head and neck carcinoma. AB - BACKGROUND: The incidence of distant metastases in head and neck cancer patients is rising because of greater locoregional control of the disease. METHODS: The relative risks for having distant metastases as first site of failure relative to the regional lymph node involvement were determined. RESULTS: The overall incidence was 10.7%, with a clear relationship between the number of involved lymph nodes and extranodal spread on one hand, and distant spread on the other hand. The group with histopathologic presence of disease in the neck had twice as much distant metastases as did those with histopathologic absence (13.6% versus 6.9%). Patients with more than three histologically positive lymph nodes were most at risk for having distant metastases (46.8%). The presence of extranodal spread meant a threefold increase in the incidence of distant metastases, compared with patients without this feature (19.1% versus 6.7%). CONCLUSIONS: Patients with three or more positive nodes and with extranodal spread may benefit from adjuvant systemic therapy. PMID- 8422639 TI - Hodgkin disease, nodular sclerosis type. Implications of histologic subclassification. AB - BACKGROUND: The prognostic significance of the cellular composition of the nodules of Hodgkin disease, nodular sclerosis type (HDNS), is controversial. METHODS: Tumors from 79 patients with HDNS, who had a median follow-up time of 9.3 years, were studied. RESULTS: Based on British National Lymphoma Investigation criteria, 58 cases were classified as NSI (low-grade) and 21 as NSII (high-grade). The study included 24 male and 55 female patients, aged 10-57 years (mean, 27 years), who presented with Stage I (13 patients [12A, 1B]), Stage II (45 patients [40A, 5B]), or Stage III (21 patients [16A, 5B]) disease. Fifty three patients had no relapse, 4 died of other causes, and 49 are in complete clinical remission. Twenty-six patients had progression of disease during therapy or relapsed and 17 were successfully salvaged. Overall length of survival was significantly shorter with NSII (P = 0.0001), extensive necrosis (P = 0.0034), high stage (P = 0.0058), and B symptoms (P = 0.030). Multivariate analysis showed that grade had the strongest effect on overall survival (P = 0.0042; hazard ratio = 10.19). The 5-year survival was 100% for NSI patients and 75% for NSII patients. Only B symptoms were significantly associated with risk of relapse after initial therapy (P = 0.030). For patients who relapsed, only histologic grade predicted subsequent disease-free survival (P = 0.0023; hazard ratio = 26.5). Five-year disease-free survival after first relapse was 94% for NSI patients and 11% for NSII patients. CONCLUSIONS: Patients with NSI disease who relapse have a more successful salvage and longer period of survival than do those with NSII disease. Histologic subclassification of HDNS appears clinically relevant, and consideration of histologic subtype may be important when planning therapy. PMID- 8422640 TI - A VM 26-based regimen for patients with previously untreated non-Hodgkin lymphoma. Prolonged disease-free survival in patients younger than 60 years of age: a phase II trial of the eastern Cooperative Oncology Group. AB - BACKGROUND: The epidophyllotoxin VM 26 has been shown to have single-agent activity in patients with diffuse aggressive lymphoma. In an attempt to determine its activity in combination with other agents known to be effective in lymphoma, a Phase II trial of a novel chemotherapy regimen was conducted. METHODS: Forty two patients with Stages II, III, and IV diffuse aggressive lymphoma were treated with teniposide, doxorubicin, prednisone, cyclophosphamide, vincristine, and bleomycin (PA Ten-CPOB) as part of a Phase II trial of the Eastern Cooperative Oncology Group. Fifty-five percent of patients had Stage IV disease, 21% Stage III, and 24% Stage II. RESULTS: The overall complete response rate was 64%. Of the 27 patients who had complete response, 19 (70% [45% of the entire group]) are still alive without disease (median follow-up, 5.7 years). No patient had a follow-up time of less than 5 years. On examination of factors that were predictive of survival and relapse, it was found that age younger than 60 years was predictive of long-term survival, as 76% of patients younger than 60 years of age were alive without disease. Forty patients were evaluable for toxicity. There were four (10%) early deaths, and six patients (15%) had Grade 4 hematologic toxicity. CONCLUSIONS: This alternating combination chemotherapy regimen (PA Ten CPOB) results in a complete response rate comparable to what has been reported previously in the literature, but 45% of patients in this series demonstrated long-term disease-free survival. When patients younger than 60 years of age with follow-up times of at least 5 years were considered, disease-free survival was 76%. PMID- 8422641 TI - Ternary complex formation and reduced folate in surgical specimens of human adenocarcinoma tissues. AB - BACKGROUND AND METHODS: Various factors, including thymidylate synthase, thymidine kinase, 5-fluorouracil phosphorylation and degradation pathways, folate concentrations, and the stability of ternary complex, which influence thymidylate synthase inhibition rate of fluoropyrimidines, were studied in 87 human adenocarcinoma tissues. RESULTS: The activity of the 5-fluorouracil degradation pathway was not significantly lower than the activity of the 5-fluorouracil phosphorylation pathway. The activity of the catabolism pathway of 5-fluorouracil should be considered in human adenocarcinoma tissue during chemotherapy. On the other hand, the means plus or minus standard deviations (means +/- SD) of the concentration of 5,10-methylenetetrahydrofolate and tetrahydrofolate were 0.69 +/ 0.54 and 1.25 +/- 0.69 nM, respectively, for the adenocarcinoma tissues without previous chemotherapy. CONCLUSIONS: Because the half-life of tritium-labeled ternary complex and folate concentration in cytosol were correlated well, the differences in folate concentration among tumors must influence the dynamic equilibrium of ternary complex formation. Moreover, these results show that the ratio of 5,10-methylenetetrahydrofolate concentration to thymidylate synthase concentration influences the thymidylate synthase inhibition rate in tumor, and that the new synthesis of 5,10-methylenetetrahydrofolate and tetrahydrofolate from other endogenous reduced folates is also important in tumors with high thymidylate synthase concentrations. PMID- 8422642 TI - Pneumocystis carinii pneumonia in patients with cancer. An increasing incidence. AB - BACKGROUND: The incidence of Pneumocystis carinii pneumonia (PCP) is increasing in patients with cancer. Possible nosocomial transmission from patients with acquired immune deficiency syndrome to those with cancer has been advocated. METHODS: Retrospectively, 17 patients with cancer were reviewed who had 20 PCP episodes that occurred during a 3-year period (1988-1990). RESULTS: Twelve patients had a hematologic malignant lesion, and five had a solid tumor as their underlying disease. Cytotoxic and/or immunosuppressive drugs were used in 16 patients (94%). The clinical presentation varied from fulminant to inapparent pneumonia. Clinical improvement and survival after appropriate therapy occurred in 12 patients (70%), whereas the remaining 5 patients died within 4 weeks of trimethoprim-sulfamethoxazole treatment initiation. In two patients, pentamidine was substituted. Complications of treatment occurred in six patients (35%). When survivors were compared with nonsurvivors, there was no difference in mean age, leukocyte counts, arterial oxygen tension, or duration of symptoms before treatment. CONCLUSIONS: In this series, nosocomial transmission of PCP was unlikely. Prophylaxis after a first episode of PCP should be considered in patients who will remain immunosuppressed. PMID- 8422643 TI - The effects of cyclooxygenase inhibitors on tumor-induced anorexia in rats. AB - BACKGROUND: Interleukin-1 (IL-1) and tumor necrosis factor (TNF) are potent induces of prostaglandin (PG) synthesis and injection of PGE, IL-1, or TNF decreases food intake in healthy animals, whereas the anorexigenic effects of injected IL-1 and TNF are blocked by inhibitors of PG synthesis. It has been hypothesized that host secretion of IL-1 and TNF contribute to tumor-induced anorexia. This study was undertaken to determine whether administration of PG inhibitors alters food intake in anorectic rats implanted with Walker 256 carcinoma. METHODS: Groups of six tumor-bearing rats were implanted with slow release pellets containing ibuprofen, indomethacin, or acetylsalicylic acid. Food intake, tumor growth, and body temperature were monitored for 14 days and compared with control tumor-bearing animals implanted with placebo pellets. RESULTS: Tumor growth was associated with anorexia, fever, weight loss, and increased leukocyte secretion of IL-1 and TNF. Indomethacin and ibuprofen retarded tumor growth 30-40% and lowered body temperature compared with controls, but had no effect on food intake or body weight of tumor-bearing animals. CONCLUSIONS: Prostaglandins do not mediate tumor-induced anorexia. PMID- 8422644 TI - The syndrome of 5-fluorouracil cardiotoxicity. An elusive cardiopathy. AB - BACKGROUND: A case of reversible cardiogenic shock linked to 5-fluorouracil (5 FU) was observed. Recognizing the increasing use of 5-FU, the authors tried to map this syndrome. METHODS: They reviewed 134 additional case reports, retrieved information from literature searches, focused on clinical features, and discussed possible pathophysiologic findings and prevention of this syndrome. RESULTS: Although angina and electrocardiographic changes were common and reproducible (approximately 90% each), coronary artery disease was found in a few patients. A total of 33 patients had severe left ventricular dysfunction, 28 without evidence of myocardial infarction. The symptoms were responsive to conservative management (90%). CONCLUSIONS: Cardiac toxicity is a little known complication of 5-FU therapy, with an unknown but significant incidence. It is highly treatable. PMID- 8422645 TI - Reduction of aberrant crypt foci induced in rat colon with azoxymethane or methylnitrosourea by feeding cholic acid. AB - Recent studies in our laboratory have demonstrated that feeding cholic acid (CHA) to rats treated with a single dose of azoxymethane (AOM) reduces the growth of putative preneoplastic lesions, aberrant crypt foci (ACF), in a dose-dependent manner. This finding was unexpected since CHA has been reported to promote colon cancer in rats receiving multiple treatments of the colon carcinogen, methylnitrosourea (MNU). The main objective of the present investigation was to evaluate the effect of the type of carcinogen and treatment protocol on the induction and growth of ACF in conjunction with CHA treatment. Male Sprague Dawley rats received 0, 1 or 2 treatments with AOM or MNU and were fed either the AIN-76A or AIN-76A plus 0.2% CHA diet for 4 weeks. The total number and average size of ACF were significantly reduced in CHA-fed animals regardless of the type or number of treatments of carcinogen. The greatest reduction of ACF due to CHA feeding was seen in the distal colon. The average crypt multiplicity (number of crypts in each ACF) was not altered by diet or carcinogen treatment. Colonic cell proliferation (crypt height and number of mitotic figures) was significantly increased in CHA-fed animals compared to control diet animals. Therefore, feeding CHA for 4 weeks reduced the number and size of ACF in rats induced by 1 or 2 injections of AOM or MNU, despite stimulation of colonic cell proliferation. These findings suggest further investigation is needed to understand the mechanism of promotion by cholic acid and the value of number and growth characteristics of ACF as a biological endpoint in the pathogenesis of colon cancer. PMID- 8422646 TI - The cellular composition in the peritoneal cavity and the cytotoxic function of the peritoneal cells from patients with ovarian cancer; effect of tumor necrosis factor-alpha treatment. AB - In patients with epithelial ovarian cancer (EOC), the cellular composition in the peritoneal cavity and the functional capacities of the peritoneal cells (PC) are unknown. Especially the peritoneal macrophages (m phi) could play an important role in defense against tumor cells. To study the cellular composition in the peritoneal cavity and the functional capacities of PC, these cells were obtained from three patients with EOC. The PC were immunophenotyped and tested functionally in vitro in a cytotoxicity assay. One of the patients was treated intraperitoneally (i.p.) with a single dose of 0.06 mg/m2 tumor necrosis factor alpha (TNF-alpha). PC were obtained before the treatment, after 24 h and after 1 week. PC from healthy women undergoing laparoscopic sterilization served as controls. It appeared that patients with EOC have a lower percentage of macrophages (m phi) in the peritoneal cavity than healthy persons. These m phi of patients were also less capable of killing U 937 tumor cells as compared to the peritoneal m phi of control persons. However, in the patient treated i.p. with TNF-alpha the cytotoxic capacities of the peritoneal m phi were strongly improved. The percentage cytotoxicity at an effector to target ratio of 10, increased from 17% to 80%. Thus, the peritoneal m phi in this patient were activated in vivo to a tumoricidal state. These findings indicate that PC in patients with EOC differ from controls, but further investigation is necessary to define the contribution of the disease and/or prior chemotherapy to this defect. PMID- 8422647 TI - Transforming growth factor-beta 2 is an autocrine growth inhibitory factor for the MOSER human colon carcinoma cell line. AB - The MOSER human colon carcinoma cell line is significantly growth inhibited by exogenous transforming growth factor-beta (TGF-beta). The secretion of TGF-beta by these cells was examined to determine if endogenous TGF-beta might also regulate MOSER cell growth. MOSER cells secreted 11 ng TGF-beta/10(6) cells, 24% of which was in the active form. Blocking antibodies specific for TGF-beta 2 stimulated growth 1.4-fold, while TGF-beta 1 specific antibodies were without effect. Treatment of MOSER cells with the differentiation agent, N,N dimethylformamide (DMF), inhibited cell growth and resulted in an 8-fold increase in secreted TGF-beta (20% active). Only antibodies specific for TGF-beta 2 were able to reverse the growth inhibitory effect of DMF on these cells. Therefore, TGF-beta 2 acted as a negative autocrine inhibitory factor for MOSER cells and the growth inhibitory effects of DMF were mediated by the increased secretion of active TGF-beta 2. PMID- 8422648 TI - Detoxication products of the carcinogenic azodye Sudan I (solvent yellow 14) bind to nucleic acids after activation by peroxidase. AB - The C-hydroxyderivatives of the carcinogenic dye Sudan I, 1-phenylazo-2,6 dihydroxynaphthalene and 1-(4-hydroxyphenylazo)-2-hydroxynaphthalene, which are considered to be detoxication products of this dye bind to DNA or tRNA after oxidation into active metabolites by peroxidase and H2O2 in vitro. The 32P postlabeling analysis of DNA modified by active metabolites of both Sudan I derivatives provides evidence that the covalent binding to DNA is the principal type of DNA modification. Since the urinary bladder is rich in peroxidases, the participation of these enzymes in activation of detoxicating products of Sudan I may be involved in the initiation of Sudan I-carcinogenesis in this organ. PMID- 8422649 TI - Human glutathione S-transferase mu (GST mu) deficiency as a marker for the susceptibility to bladder and larynx cancer among smokers. AB - The isoenzyme mu of glutathione S-transferase (GST mu) is dominantly inherited and the prevalence of this isoenzyme in the population is about 60%. An increased risk of lung cancer has been previously shown among smokers lacking GST mu in (Seidegard J., Pero R.W., Miller D.G., Beattie E.J. (1986) Carcinogenesis, 7, 751 753). The frequency of the phenotypes of this isoenzyme in bladder cancer patients (n = 75), in larynx cancer patients (n = 78) and healthy controls matched for age and smoking history is reported here. A significantly higher proportion of smokers in the control group had measurable GST mu compared with bladder cancer patients (54.6% vs. 33.3%, P < 0.01) and also compared to larynx cancer patients (55.1% vs. 33.3%, P < 0.01). Odds ratio analysis indicates that smokers with this polymorphic variant have an approximately 2-fold greater risk of developing these cancers. PMID- 8422650 TI - Enhancement of tumorigenesis by N-nitrosodiethylamine, N-nitrosopyrrolidine and N6-(methylnitroso)-adenosine by ethanol. AB - Inclusion of 10% ethanol with 6.8 ppm N-nitrosodiethylamine in the drinking water of strain A male mice resulted in a 4-fold enhancement of multiplicity of lung tumors and a 16-fold increase in incidence of fore-stomach tumors, compared with carcinogen alone. Given with 40 ppm N-nitrosopyrrolidine, ethanol caused a 5.5 fold increase in lung tumor multiplicity. The inclusion of 15% ethanol with N6 (methylnitroso)adenosine, given orally to Swiss female mice, led to reduced body weights and shortened survival time related to hemangiosarcoma occurrence or increased incidence of thymic lymphoma, depending on dose of carcinogen. The data provide additional support for the proposal that co-administered ethanol increases the tumorigenicity of nitrosamines by blocking hepatic first-pass clearance. PMID- 8422651 TI - Effect of restricted caloric intake on the development of the azoxymethane induced glutathione S-transferase placental form positive hepatocellular foci in male F344 rats. AB - The modifying effect of 30% caloric restriction on the occurrence of azoxymethane (AOM)-induced glutathione S-transferase placental form (GST-P) positive hepatocellular foci was investigated in male F344 rats. Starting at 5 weeks of age, groups of animals were fed ad libitum a high-fat (23.5%) semipurified diet. At 7 weeks of age, all animals except the vehicle-treated groups were s.c. injected with AOM (15 mg/kg body wt., once weekly for 2 weeks). Four days after the second injection, groups of animals were continued on high-fat diet and fed ad libitum (ad libitum group) whereas other groups were restricted to 70% of total calories (calorie-restricted group) consumed by the ad libitum group, but received the same amounts of fiber, vitamins and minerals. Thirty-two weeks after AOM injections, all animals were necropsied and livers were sectioned and stained for GST-P by a immunohistochemical technique for quantitative analysis of enzyme altered foci of the liver. Comparing AOM treated groups. The density and the unit area of enzyme altered foci were significantly lower in the calorie-restricted group (3.84 +/- 1.55/cm2, 7.96 +/- 5.43%) than in the ad libitum group (10.14 +/- 3.62/cm2, 28.11 +/- 12.33%). The size of foci was also reduced in the calorie restricted group (17.15 x 10(-3) mm2 vs. 32.36 x 10(-3) mm2). The incidence and density of hepatocellular foci in rats fed calorie restricted diet were significantly lower than those in rats fed ad libitum, comparing vehicle-treated groups. These results indicate that calorie restriction inhibited the occurrence of both of spontaneous and AOM induced GST-P positive foci in rats. PMID- 8422652 TI - Modulation of cytochrome P-450IA1-mediated mutagenicity, DNA binding and metabolism of benzo[a]pyrene by Chinese medicinal herbs. AB - Oldenlandia diffusa (OD) and Scutellaria barbata (SB) have been used in traditional Chinese medicine for treating liver, lung and rectal tumors. We previously showed that they inhibited mutagenesis, DNA binding and metabolism of benzo[a]pyrene (BaP) and aflatoxin B1 (AFB1) bioactivated by Aroclor 1254-induced rat hepatic S9. The purpose of this study was to investigate the effects of OD and SB on the cytochrome P-450IA1-mediated mutagenicity of BaP in Salmonella typhimurium TA100 using beta-naphthoflavone (beta NF)-induced rat hepatic S9. We also determined the effects of OD and SB on cytochrome P-450IA1-linked ethoxyresorufin O-deethylase (EROD) activity in beta NF-induced hepatic microsomes. In addition, we studied the effects of these two herbs on BaP metabolite binding to calf thymus DNA and using high performance liquid chromatography (HPLC) we investigated the effects of OD and SB on the metabolism of BaP by beta NF-induced S9. Our experimental results showed that OD and SB inhibited the mutagenicity of BaP in the presence of either non-induced or beta NF-induced S9. SB significantly inhibited BaP binding to DNA. These effects correlated with the inhibition of cytochrome P-450IA1-linked EROD activity in beta NF-induced microsomes and with an inhibition of beta NF-induced S9 mediated metabolism of [3H]BaP as determined by HPLC. These results suggest that OD and SB may possess antimutagenic activity by inhibiting P-450IA-mediated metabolism of BaP. PMID- 8422653 TI - High incidence of mutations of the p53 gene detected in ovarian tumours by the use of chemical mismatch cleavage. AB - We have investigated a series of ovarian tumours for evidence of mutations in the p53 tumour suppressor gene. In this study we have made use of the chemical mismatch cleavage technique which, from analyses of other genes, has been shown to consistently identify all point mutations present within a region of DNA. This approach revealed mutations of p53 in 11/20 tumours studied, mainly in exons 5 or 7. After sequencing the relevant regions of the gene it was shown that ten of these mutations would have resulted in an amino acid substitution in the protein and only one represented a polymorphism. The observed incidence of p53 missense mutations in our series (50%) was the highest recorded in ovarian tumours and demonstrated the potential of the mismatch cleavage technique as a reliable method for the detection of p53 mutations in human tumours. PMID- 8422654 TI - Hypersensitivity vasculitis and systemic lupus erythematosus induced by anticonvulsants. AB - Vasculitis can be a systemic manifestation of hypersensitivity to many drugs, among them anticonvulsants. The clinical manifestations include rash and renal, hepatic, and pulmonary involvement. Diagnosis is based upon clinical findings and a characteristic biopsy showing granulocytic and sometimes eosinophilic infiltrates around small blood vessels, especially venules. A severe form of hypersensitivity vasculitis, with extensive visceral involvement and poor prognosis, has been encountered very rarely following phenytoin and in isolated cases following carbamazepine and trimethadione administration. Drug-induced systemic lupus erythematosus is much more frequent, with distinct clinical and laboratory abnormalities. The syndrome was described following treatment with most anticonvulsants in clinical use--phenytoin, carbamazepine, ethosuximide, trimethadione, primidone, and valproate, but not phenobarbital or benzodiazepines. The early recognition of these syndromes as being related to drugs is important, because they usually remit upon withdrawal of the offending agent. PMID- 8422655 TI - Is there a relationship between Parkinson's disease and essential tremor? AB - The issue of whether there is a relationship between essential tremor (ET) and Parkinson's disease (PD) is controversial partly due to the confusion regarding the accurate diagnosis of these conditions. The presence of postural tremor, which often occurs in PD, by itself is insufficient for the diagnosis of ET. Most epidemiological studies have shown that there is no association between these two conditions. Some studies, but not others have found a higher prevalence of tremor in family members of PD patients. Clinical, positron emission tomography scan, and neuropathological data have failed to show any relationship between these two conditions. It is concluded that there is no relationship between ET and PD and that when these two conditions are seen in the same patient, this represents a chance occurrence of two common diseases. PMID- 8422656 TI - Quantitative assessment of parkinsonian patients by continuous wrist activity monitoring. AB - The aim of this study was to examine the quantitative relationship between activity monitor measures and clinical scores of patients with Parkinson disease (PD). Motor activity was recorded continuously for 5 to 6 days at home with a wrist-worn activity monitor in 69 PD patients and 59 healthy controls. Clinical scores of the patients, age, and sex were submitted to multiple regression analysis to examine the quantitative relationship with measures reflecting the activity level and the proportion of activity and immobility over time. The patients' age, sex, and scores representing hypokinesia and rigidity and resting tremor explained approximately 50% of the variance of the motor activity measures. All motor activity measures declined with age; the rate of decline was similar for the patients and controls. Sex emerged as a predictor of the motor activity measures in the patients only. Male patients with PD showed significantly lower values for all motor activity measures than female patients and controls. Our results show that activity monitoring can be used as an objective quantitative assessment in PD. PMID- 8422657 TI - Short-term beneficial effect of deprenyl monotherapy in early Parkinson's disease: a quantitative assessment. AB - It was recently shown that early treatment with deprenyl in patients with Parkinson's disease can delay the need for initiation of levodopa therapy. It was therefore suggested that deprenyl may slow down disease progression. Alternatively, the observed stabilization of clinical disability may merely reflect drug-induced symptomatic benefit. We therefore examined a possible short term beneficial effect of deprenyl (10 mg/day) as the first and only drug in 15 consecutive de novo patients. Bradykinesia was quantitatively assessed by computerized analysis of isometric force/time curves of biceps and triceps bilaterally and by calculation of simple reaction time (RT) and maximal muscle contraction rate (MMCR). We also measured maximal muscle force, evaluated clinical status by the Unified Parkinson's Disease Rating Scale (UPDRS) motor score, and recorded patients' subjective reports. All tests were carried out before and at 1 and 2 months of treatment. Only one patient reported a beneficial effect. No significant changes in the UPDRS or score or muscle force were observed. In contrast, MMCR and RT improved at 1 month by an average of 12.1 and 7.2%, respectively (p < 0.01, paired t test). This improvement persisted after 2 months of treatment. Study shows that deprenyl monotherapy exerts a short-term beneficial effect in de novo parkinsonian patients. This effect, however, appears to be small, subclinical, and probably cannot account for the observed delay in the need to start levodopa therapy. PMID- 8422658 TI - Safety and efficacy of oral physostigmine in the treatment of Alzheimer disease. AB - Results of therapeutic trials with physostigmine in the treatment of Alzheimer disease (AD) have been inconsistent and controversy persists concerning safety and efficacy. In a double-blind, placebo-controlled, crossover study, patients received 6 weeks of oral physostigmine (OP) and placebo in random order. Twenty nine patients with AD received as much as 16 mg/day of OP and were assessed with neuropsychological and functional measures. No significant cardiac side effects were noted, though other systemic adverse effects were noted, requiring dose reduction in four patients. There was a slight but significant improvement (12%) in performance on the selective reminding test with physostigmine and the memory performance was correlated with dosage. This improvement compares favorably with the 15% decrease in scores seen in an untreated comparison cohort followed for an equivalent time period. There was a trend toward an improvement in communication and a reduction in memory complaint. These results suggest that oral physostigmine is safe and may improve memory in AD. PMID- 8422659 TI - Effects of yohimbine on plasma catecholamine levels in orthostatic hypotension related to Parkinson disease or multiple system atrophy. AB - Different pathophysiological mechanisms may underly orthostatic hypotension (OH) observed in neurological degenerative disorders. The present study investigates the responses to the pharmacological activation of sympathetic pathways induced by yohimbine (0.2 mg/kg orally) through measurements of plasma catecholamine levels in parkinsonian patients with (n = 9) or without OH (n = 11), in patients with multiple system atrophy (MSA) plus OH (n = 9), and in controls (n = 6). Basal norepinephrine plasma levels in parkinsonian patients with OH (71 +/- 11 pg/ml) were significantly lower (p < 0.05) than in parkinsonian patients without OH (280 +/- 25 pg/ml) or in controls (259 +/- 48 pg/ml). In patients with MSA plus OH, basal catecholamine plasma levels were in the normal range (344 +/- 54 pg/ml). Yohimbine significantly increased norepinephrine (p < 0.05) but not epinephrine plasma levels in all groups. However, the increment obtained in parkinsonian patients with OH (+53 +/- 18 pg/ml) remained significantly lower (p < 0.05) than in parkinsonian patients without OH or in controls (+638 +/- 140 and +457 +/- 103 pg/ml, respectively) as well as in MSA plus OH (+633 +/- 142 pg/ml). Yohimbine failed to modify the blood pressure and heart rate at the dose used. The results suggest that the yohimbine test is useful to elucidate the site of the dysfunction of the efferent sympathetic pathways in these two conditions. In Parkinson disease with OH, the lesion is both central and postganglionnic, whereas in MSA it is only centrally located. PMID- 8422660 TI - Pharmacokinetic comparison of two albendazole dosage regimens in patients with neurocysticercosis. AB - To evaluate two different dosage regimens for albendazole (7.5 mg/kg twice a day or 5.0 mg/kg three times a day), a study was performed in 10 patients with a diagnosis of parenchymal brain cysticercosis. Each patient received both regimens sequentially according to a randomized, crossover design. Blood and urine samples were taken once the drug steady state had been reached. Plasma levels of albendazole sulfoxide at steady state were determined using a HPLC method. In spite of a great intersubject variability observed with both regimens in the area under the curve (AUC) and in the minimum steady-state plasma concentration (Cp min ss), no statistically significant differences were found in these parameters. We suggest that a regimen of 7.5 mg/kg every 12 h can favorably replace the currently used regimen of 5 mg/kg every 8 h. PMID- 8422661 TI - Effects of selegiline dosing on motor fluctuations in Parkinson's disease. AB - To examine selegiline's dosing effects, we studied 16 Parkinson disease patients with motor fluctuations in a double-blind, crossover trial of selegiline at 0, 5, and 10 mg daily. Patients' motor diaries, disability scores, and walking speed were marginally improved, with no advantage of 10 mg over 5 mg found. We conclude that in patients with motor fluctuations, selegiline provides modest symptomatic effects, with daily dosing of 5 mg being indistinguishable from 10 mg. PMID- 8422662 TI - Ketorolac in reflex sympathetic dystrophy. AB - Treatment of reflex sympathetic dystrophy (RSD) is primarily surgical. Typically, regional sympathetic blockade or sympathectomy is employed. Alternatives to neurosurgery, such as pharmacotherapy, include systemic corticosteroids and antidepressants. However, side effects and nonresponders to these approaches are not uncommon. Recently, ketorolac was demonstrated to relieve pain in RSD patients when administered by intravenous regional block (IVRB), yet this method has obvious limitations in the outpatient setting. The following case demonstrates marked improvement in treatment-resistant RSD pain with intramuscular ketorolac. PMID- 8422663 TI - Specific immune recognition of autologous tumor by lymphocytes infiltrating colon carcinomas: analysis by cytokine secretion. AB - Tumor-infiltrating lymphocytes (TIL) were grown in the presence of interleukin-2 from 19 colon carcinoma specimens, including 1 primary lesion and 18 metastatic lesions. These cultures showed a median proliferation of 606-fold (range 13-fold to 28,000-fold) over 49 culture days (range 26-76 days). By phenotype, mature cultures were 69%-99% CD3+ (mean 93%) and contained mixed populations of CD4+ and CD8+ cells (CD4 > CD8 in 10 of 19 cultures). Fresh cryopreserved colon tumors were not lysed by autologous TIL in short-term 51Cr-release assays, and were poorly lysed by lymphokine-activated killer cells. Ten TIL cultures were assayed for cytokine secretion in response to autologous and allogeneic tumors during a 6 to 24-h coincubation. Culture supernatants were tested by ELISA for the presence of granulocyte/macrophage-colony-stimulating factor, interferon gamma, and tumor necrosis factor alpha. Of 10 TIL, 4 secreted at least two of these cytokines specifically in response to autologous and/or HLA-matched fresh allogeneic colon carcinomas, but not to melanomas or HLA-unmatched colon carcinomas. Cytokine secretion was mediated by both CD4+ and CD8+ TIL, and could be inhibited by mAb directed against the appropriate class of MHC antigen. These data provide evidence for specific, MHC-restricted immune recognition of human colon carcinomas by T lymphocytes. PMID- 8422664 TI - Effect of cisplatin upon expression of in vivo immune tumor resistance. AB - The major intent of cancer treatment with cytotoxic drugs is direct tumor cell damage, but some of these drugs have been shown to be immunomodulatory. Cisplatin is a widely used cytotoxic drug that has been combined with biological response modifiers in recent clinical trials. To evaluate further whether cisplatin may independently alter the level of host resistance against tumor growth, the drug was tested in the Mc7 sarcoma rat tumor model. The expression of in vivo tumor resistance against Mc7 sarcoma in syngeneic Wistar rats is mediated by circulating non-cytotoxic T lymphocytes. These cells interact specifically with tumor cells to generate cytotoxic effectors locally at the site of a tumor challenge. Activities of these components of expression of tumor resistance were measured in vivo after administration of cisplatin and dose-dependent effects were found. Low-dose cisplatin (0.3 mg/kg) increased the activity of the circulating lymphocytes that mediate tumor resistance, and high-dose cisplatin (9 mg/kg) suppressed both mediator lymphocyte activity and the generation of antitumor effector mechanisms. These studies suggest that low-dose cisplatin may be immunomodulatory and combining it with biological response modifiers might be a useful strategy. However, high-dose cisplatin given with biological response modifiers may negate potential immunomodulatory activities of such agents. PMID- 8422665 TI - Effects of natural interferon alpha, natural tumor necrosis factor alpha and their combination on human mesothelioma xenografts in nude mice. AB - Effects of human natural interferon alpha (nIFN) alone, human natural tumor necrosis factor alpha (nTNF) alone and their combination (OH-1) were tested on three human mesothelioma lines implanted in nude mice. Tumors were transplanted subcutaneously by trocar on treatment day -12. nIFN was given intraperitoneally (i.p.) at a dose of 2 x 10(7) or 2 x 10(8) IU kg-1 day-1, 5 days a week for 3 weeks. nTNF was given i.p. at a dose of 2 x 10(7) or 2 x 10(8) U kg-1 day-1 in the same schedule as that of nIFN. Tumor diameters were serially measured and tumor volumes were calculated. Antitumor effects were assessed by two methods: comparison of final tumor volumes in treated and control groups (T/C), and changes in median average total tumor volume. The treatment produced no clinically discernible toxicities. nIFN had strong inhibitory activity against all three human mesothelioma lines. nTNF alone had modest activity only at the high dose used. The combination of the two produced activity essentially similar to that produced by nIFN alone. High-dose nIFN may have a role as an active agent in the treatment of patients with mesothelioma. PMID- 8422666 TI - Characterization of the exogenous interleukin-2 requirements for the generation of enhanced antitumor cytotoxicity by thymocytes from low-dose melphalan-treated MOPC-315 tumor bearers. AB - We have shown previously that thymocytes from MOPC-315-tumor-bearing mice treated with low-dose melphalan (L-phenylalanine mustard) (L-PAM TuB mice) are superior to thymocytes from untreated MOPC-315-tumor-bearing mice or thymocytes from untreated normal mice or normal mice treated with low-dose melphalan in their ability to generate an antitumor cytotoxic response following 5-day in vitro stimulation with MOPC-315 tumor cells in the presence of a low concentration of recombinant interleukin-2 (rIL-2) [Mokyr MB, Bartik MM, Ahn M-C (1989) Cancer Res 49; 870]. Here we characterize the rIL-2 requirements for the generation of enhanced antitumor cytotoxicity by L-PAM TuB thymocytes relative to normal thymocytes upon in vitro stimulation with MOPC-315 tumor cells. Specifically, we show that delaying the addition of a low concentration of rIL-2 to 5-day in vitro stimulation cultures of thymocytes resulted in a progressive decline in the generation of antitumor cytotoxicity by both normal and L-PAM TuB thymocytes. However, even when rIL-2 was added on day 2 after culture initiation, thymocytes from L-PAM TuB mice generated a more potent antitumor cytotoxicity than did thymocytes from normal mice. In addition, when rIL-2 was added at the time of culture initiation, replacement of the conditioned medium with fresh medium lacking rIL-2 on day 3 of the 5-day in vitro stimulation culture period eliminated the ability of normal thymocytes, and reduced (but did not eliminate) the ability of L-PAM TuB thymocytes, to generate a significant level of antitumor cytotoxicity. A low concentration of fresh rIL-2 was sufficient to restore completely the generation of antitumor cytotoxicity by normal or L-PAM TuB thymocytes when added to the stimulation cultures immediately after the removal of the rIL-2-containing conditioned medium. The same low concentration of rIL-2 was also sufficient for restoring the generation of antitumor cytotoxicity by cultures of L-PAM TuB thymocytes, but not normal thymocytes, from which the rIL-2 containing medium was removed 1 day earlier. At the same time, conditioned medium from stimulation cultures of L-PAM TuB thymocytes was not superior to conditioned medium from stimulation cultures of normal thymocytes in supporting the generation of antitumor cytotoxicity by either normal or L-PAM TuB thymocytes.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8422667 TI - Effect of ibuprofen on monocyte activation by liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (CGP 19835A): can ibuprofen reduce fever and chills without compromising immune stimulation? AB - The purpose of this study was to determine the effects of ibuprofen on the ability of liposome-encapsulated muramyl tripeptide phosphatidylethanolamine (L MTP-PE) to activate human blood monocytes in vitro. We undertook these experiments because the major toxic side-effects following L-MTP-PE infusion, fever and chills, could be prevented when ibuprofen was given orally immediately before L-MTP-PE infusion. It was therefore important to determine whether ibuprofen interfered with the macrophage-activation properties of L-MTP-PE. Peripheral blood monocytes were isolated from normal donors, then incubated with L-MTP-PE in the presence or absence of ibuprofen. The cytotoxic properties of the monocytes were assessed by a radioisotope-release assay against A375 cells. Ibuprofen at dose levels of 40 micrograms/ml suppressed the generation of the cytotoxic phenotype but did not interfere with the killing process once the cells were activated. Interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF alpha) production, as well as the mRNA expression of these cytokines, was suppressed by 40 micrograms/ml ibuprofen. Since IL-1 and TNF play a crucial role in the cytotoxic function of monocytes, these findings may explain the mechanism by which ibuprofen inhibited the generation of the cytotoxic phenotype by L-MTP-PE. By contrast, ibuprofen dose levels up to 10 micrograms/ml had no effect on the generation of monocyte-mediated cytotoxicity by L-MTP-PE and no effect on the production, secretion, or mRNA expression of TNF and IL-1. Therefore, we concluded that if ibuprofen is to be used to control the side-effects of L-MTP PE, blood levels of up to 10 micrograms/ml are desirable. In two of three patients, we determined that an oral dose of 200 mg given immediately before L MTP-PE infusion could achieve these desired blood levels. PMID- 8422668 TI - Clinical application of recombinant human erythropoietin for treatments in patients with head and neck cancer. AB - The therapeutic effects of intravenous recombinant human erythropoietin (r-hEPO) administration on anemia induced by radiation therapy (3 cases), chemotherapy (18 cases) and combined therapies (5 cases) in patients with head and neck malignancies were examined. The effectiveness was evaluated by the changes in the hemoglobin concentration examined before and after the r-hEPO administration. The r-hEPO administration combined with anticancer therapies improved anemia induced by all three treatments. The therapeutic effectiveness of r-hEPO injection was also noted on anemia induced by all of four different chemotherapeutic regimens that have been ordinarily used for head and neck malignancies. Furthermore, the efficacy of the different dose schedules, 3000 IU (12 cases) or 6000 IU (14 cases), three times a week, was compared. Both of the r-hEPO dose schedules were effective for anemia, but the efficacy of 6000 IU was superior to that of 3000 IU. No significant changes were observed in the number of white blood cells and platelets and the results of biochemical examinations after the r-hEPO injection. There were no objective side-effects related to the r-hEPO administration. These results suggest that anemia induced by chemotherapy and/or radiotherapy could be prevented by r-hEPO administration. The addition of r-hEPO to anticancer therapies would make it possible to pursue the planned therapeutic schedules, prevent the decrease of immunity after allogeneic blood transfusion and bring about an-improvement in the prognosis of patients with malignancies. PMID- 8422669 TI - Prognostic value of non-MHC-restricted killer cell activity in lung cancer. AB - The prognostic value of peripheral blood non-MHC-restricted cytotoxicity against the myeloid leukaemic line K562 in lung cancer patients was studied. At the time of diagnosis and before operation, 57 patients with lung cancer were tested for cytotoxicity and subsequently followed for up to 4 years. In addition, 145 lung cancer patients, 30 patients with non-neoplastic lung diseases and 76 healthy donors were tested for cytotoxicity without the follow-up, in order to correlate the stage of lung cancer and the growth rate of tumours to the level of non-MHC restricted cytotoxicity. On average, lung cancer patients had similar non-MHC restricted cytotoxicity to the controls. However, patients with stage II-IV diseases showed an impaired activity, stages III and IV differing significantly from the controls. This result shows that the decline in natural killer (NK) activity is associated with tumour burden. Patients with slowly growing neoplasms had stronger cytotoxic activity than patients with fast or moderately progressing disease. In the follow-up study, the whole material of 57 patients showed only a slight correlation between cytotoxicity and survival: 42% of the patients with strong activity survived for more than 2.5 years, whereas 6% of the patients with weak activity did so. In stage I patients there was no correlation between cytotoxicity and survival, nor was there a correlation in patients with stages II IV of the disease. Hence, in our group of patients the determination of cytotoxicity preoperatively yielded no prognostic information beyond that already available from staging. However, those stage II-IV patients that survived for 1 year or more after the diagnosis and cytotoxicity tests, showed a significant correlation between cytotoxicity and survival. PMID- 8422671 TI - Unwinding dosage compensation. PMID- 8422670 TI - Immunogenicity of synthetic peptides related to the core peptide sequence encoded by the human MUC1 mucin gene: effect of immunization on the growth of murine mammary adenocarcinoma cells transfected with the human MUC1 gene. AB - The immune response of CAF1 mice to various synthetic peptides (SP) related to the amino acid sequence (PDTRPAPGSTAPPAHGVTSA) of the tandem repeat of the MUC1 human breast mucin core peptide was evaluated. The most immunogenic preparations of the synthetic peptides were those conjugated to keyhole limpet hemocyanin (KLH) or clustered in a dendritic multiple antigenic peptide (MAP-4) configuration. The mice were immunized subcutaneously with synthetic peptides emulsified in RIBI adjuvant, employing various immunization protocols. Equivalently high IgG responses were induced using SP-KLH conjugates (GVTSAPDTRPAPGSTA-KLH) or an SP--MAP-4 chimeric configuration (SP1-6), which also included a universal malarial CST-3 T-helper epitope (SP1-6 = SAPDTRPAEKKIAKMEKASSVFNVVNS--MAP-4). These IgG antibodies bound both the appropriate MUC1 synthetic peptides and the cell surface expressed MUC1 mucin on murine mammary cells that had been transfected with the human MUC1 gene and a human breast cancer cell line that expresses cell-surface MUC1. A MAP-4 molecule, which included the entire 20-amino-acid sequence of the MUC1 tandem repeat (SP1-5 = PDTRPAPGSTAPPAHGVTSA-MAP-4) induced a poor IgG response. In contrast, all three types of molecule: SP-KLH, SP1-6 and SP1-5, were found to be good immunogens for the induction of specific delayed-type hypersensitivity (DTH) reactions measured using either synthetic peptides or MUC1-transfected cells. In addition, immunization with irradiated MUC1-transfected cells induced strong DTH reactions measured using synthetic peptides that expressed the PDTRP sequence, which has been shown to be, or to overlap, a T cell epitope in humans and a B cell epitope in mice. Finally, it was demonstrated that synthetic MUC1 peptide "vaccines" could be used both prophylactically and therapeutically to inhibit the growth of MUC1-transfected tumor cells and prolong the survival of tumor-bearing mice. PMID- 8422672 TI - A short, disordered protein region mediates interactions between the homeodomain of the yeast alpha 2 protein and the MCM1 protein. AB - Homeodomains are folded into a characteristic three-dimensional structure capable of recognizing DNA in a sequence-specific manner. We show that correct target site selection by the yeast alpha 2 protein requires, as well as its homeodomain, an adjacent short and apparently unstructured region of the protein. This flexible homeodomain extension is responsible for specifying an interaction with a second regulatory protein, MCM1, which permits the cooperative binding of the two proteins to an operator. Two additional experiments suggest that this extension-homeodomain arrangement is likely to have some generality. First, when the extension of alpha 2 is grafted onto the Drosophila engrailed homeodomain, it yields a protein with the DNA binding specificity of engrailed and the ability to bind cooperatively to DNA with MCM1. Second, the alpha 2 extension specifies interaction not only with the yeast MCM1 protein, but also with the related human protein SRF. PMID- 8422673 TI - Function of the S. cerevisiae DST1/PPR2 gene in transcription elongation. PMID- 8422674 TI - Frameshifting in the expression of the E. coli trpR gene occurs by the bypassing of a segment of its coding sequence. AB - The E. coli trpR gene encodes the 108 amino acid long trp repressor. We have previously shown that a +1 frameshifting event occurs during the expression of trpR. Here we show that the transition from the 0 to the +1 frame of trpR occurs by the bypassing of a 55 nt long segment of the trpR+1-lacZ mRNA. This bypassing event is not pretranslational, and it probably takes place during translation. Two adjacent elements are required: a specific sequence of trpR, which must be preceded by a nonspecific 5' end longer than 10 translatable codons. Unique to trpR-lacZ bypassing is that the 55 nt long region must be translated in frame 0 to enable bypassing into the +1 frame. Translational bypassing as a newly discovered mechanism of gene expression is discussed, and the possible existence of translational introns is suggested. PMID- 8422675 TI - Molecular biology of carrier proteins. PMID- 8422676 TI - The DNA-dependent protein kinase: requirement for DNA ends and association with Ku antigen. AB - The DNA-dependent protein kinase (DNA-PK) phosphorylates Sp1 and several other nuclear proteins. Here, we show that Sp1 and the DNA-PK must be colocalized on the same DNA molecule for efficient phosphorylation to occur. Interestingly, we find that the DNA-PK binds to and is activated by the ends of DNA molecules. Furthermore, we show that the DNA binding properties of the DNA-PK are identical to those of Ku, a well-characterized human autoimmune antigen. We demonstrate that the DNA-PK can be fractionated into two components, one of which is Ku and the other of which is a polypeptide of approximately 350 kd. DNA cross-linking and coimmunoprecipitation studies indicate that the catalytic 350 kd DNA-PK component is directed to DNA by protein-protein interactions with Ku. The implications of the unusual DNA binding mode and multicomponent nature of the DNA PK are discussed. PMID- 8422677 TI - DNA deletion associated with hereditary neuropathy with liability to pressure palsies. AB - Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal dominant disorder that causes episodes of focal demyelinating neuropathy following minor trauma to peripheral nerves. We assign the HNPP locus to chromosome 17p11.2 and demonstrate the presence of a large interstitial deletion associated with this disorder in three unrelated pedigrees. De novo deletion is documented in one pedigree. The deleted region appears uniform in all pedigrees and includes the gene for peripheral myelin protein 22 (PMP-22), suggesting that underexpression of PMP-22 may cause HNPP. The deletion in HNPP spans approximately 1.5 Mb and includes all markers that are known to map within the Charcot-Marie-Tooth neuropathy type 1A (CMT1A) duplication. Furthermore, the breakpoints in HNPP and CMT1A map to the same intervals in 17p11.2, suggesting that these genetic disorders may be the result of reciprocal products of unequal crossover. PMID- 8422678 TI - A role for synaptotagmin (p65) in regulated exocytosis. AB - Proteins that are specifically localized to synaptic vesicles in the nervous system have been proposed to mediate aspects of synaptic transmission. Antibodies raised against the cytoplasmic domains of five of these proteins, vamp, rab3A, synaptophysin, synaptotagmin, and SV2, were used to investigate their function. Microinjection of monoclonal and polyclonal antibodies raised against synaptotagmin (p65), but not the other vesicle proteins, decreases K+/Ca(2+) mediated dopamine beta-hydroxylase surface staining, a measure of regulated secretion in PC12 cells. Microinjection of a soluble fragment of synaptotagmin encompassing one of the domains homologous to the C2 regulatory region of protein kinase C, but lacking the membrane anchor, also inhibits evoked dopamine beta hydroxylase surface staining. These results provide support for the hypothesis that synaptotagmin, a Ca(2+)- and phospholipid-binding protein, is important for regulated exocytosis in neurons. PMID- 8422679 TI - A nuclear pore complex protein that contains zinc finger motifs, binds DNA, and faces the nucleoplasm. AB - We have molecularly cloned and sequenced a cDNA for a rat liver nucleoporin with a molecular mass of 152.8 kd, termed nup153, that shares a repetitive degenerate pentapeptide motif with a subgroup of nucleoporins of yeast and vertebrates. However, its most striking feature is a novel 4-fold repeat of a Cys2-Cys2-type zinc finger motif. When expressed in E. coli, the zinc finger domain of nup153 binds DNA in a zinc-dependent fashion. Immunoelectron microscopy localized nup153 exclusively to the nucleoplasmic side of the nuclear pore complex. We suggest that nup153 recognizes a specific DNA sequence to organize the genome three dimensionally and to gate transcribable genes to nuclear pore complexes. PMID- 8422680 TI - Determination of cleavage planes. PMID- 8422681 TI - Regulation of the sex-specific binding of the maleless dosage compensation protein to the male X chromosome in Drosophila. AB - In Drosophila, the single male X chromosome is transcribed at twice the rate of a single female X chromosome. This hypertranscription requires the functions of at least four autosomal male-specific lethal genes (msls) and is under the control of the Sex-lethal (Sxl) gene. One of the msls, the maleless (mle) gene, encodes a protein that is associated with the male X chromosome. To investigate how dosage compensation is regulated, we have determined whether Sxl and the other msls are required for mle X chromosome binding. We have found that in females, Sxl functions to prevent mle from binding to the two X chromosomes. Additionally, we have found that mle X chromosome binding requires wild-type msl1, msl2, and msl3 functions. These data support a model whereby the activity of the mle protein is regulated through its association with one or more of the other msl proteins. PMID- 8422682 TI - Saccharomyces telomeres acquire single-strand TG1-3 tails late in S phase. AB - Saccharomyces telomeres consist of approximately 300 bp of C1-3A/TG1-3 DNA. Nondenaturing Southern hybridization, capable of detecting approximately 60 to approximately 300 bases of TG1-3 DNA, revealed that yeast telomeres acquired and lost TG1-3 tails, the predicted intermediate in telomere replication, in a cell cycle-dependent manner. TG1-3 tails were also detected on the ends of a linear plasmid isolated from late S phase cells. In addition, a nonlinear form of this plasmid was detected: this structure migrated in two-dimensional agarose gels like a nicked circle of the same size as the linear plasmid, but had considerably more single-stranded character than a conventional nicked circle. The evidence indicates that these circles were formed by telomere-telomere interactions involving the TG1-3 tails. These data provide evidence for a cell cycle-dependent change in telomere structure and demonstrate that TG1-3 tails, generated during replication of a linear plasmid in vivo, are capable of mediating telomere telomere interactions. PMID- 8422683 TI - RTG1 and RTG2: two yeast genes required for a novel path of communication from mitochondria to the nucleus. AB - The expression of some nuclear genes is sensitive to the functional state of mitochondria, a process we term retrograde regulation. Here we show that retrograde regulation of the yeast CIT2 gene encoding peroxisomal citrate synthase depends on a new class of upstream activation site element (UASr) and two previously unidentified genes, RTG1 and RTG2. RTG1 encodes a protein of 177 amino acids with similarity to basic helix-loop-helix transcription factors that likely functions at the CIT2 UASr. RTG2 encodes a protein of 394 amino acids of unknown function. Cells containing null alleles of RTG1 and RTG2 are viable and respiratory competent. However, they are auxotrophic for glutamic or aspartic acid and cannot use acetate as a sole carbon source, suggesting that both the tricarboxylic acid and glyoxylate cycles are compromised. Thus, RTG1 and RTG2 are pivotal genes in controlling interorganelle communication between mitochondria, peroxisomes, and the nucleus. PMID- 8422684 TI - Three in one and one in three: it all depends on TBP. PMID- 8422685 TI - A positive role for histone acetylation in transcription factor access to nucleosomal DNA. AB - Acetylation of the N-terminal tails of the core histones directly facilitates the recognition by TFIIIA of the 5S RNA gene within model chromatin templates. This effect is independent of a reduction in the extent of histone-DNA interactions or a change in DNA helical repeat; it is also independent of whether a histone tetramer or octamer inhibits TFIIIA binding. Removal of the N-terminal tails from the core histones also facilitates the association of TFIIIA with nucleosomal templates. We suggest that the histone tails have a major role in restricting transcription factor access to DNA and that their acetylation releases this restriction by directing dissociation of the tails from DNA and/or inducing a change in DNA configuration on the histone core to allow transcription factor binding. Acetylation of core histones might be expected to exert a major influence on the accessibility of chromatin to regulatory molecules. PMID- 8422686 TI - The adenovirus protease is activated by a virus-coded disulphide-linked peptide. AB - In common with many other viruses, adenoviruses code for a protease essential for the development of infectivity. Recombinant adenovirus protease was active in crude in vitro complementation assays but was inactive with peptide or purified protein substrates. Activity was reconstituted by a component of adenovirus virions, which was identified as GVQSLKRRRCF, a peptide derived from the virus protein pVI. Synthetic peptides were used to demonstrate that the cysteine is essential and that the disulphide-linked dimer is required for activity. It is proposed that the adenovirus protease is a cysteine protease and that its activation by the peptide involves thiol-disulphide interchange, which serves to expose the active site cysteine. This represents a novel strategy for controlling the activity of a protease that is required for virus maturation. PMID- 8422687 TI - Quantitation of the synergistic interaction of edatrexate and cisplatin in vitro. AB - Cytotoxicity studies combining edatrexate (EDX) and cisplatin (Cis-Pt) were carried out in HL-60 cells in vitro as a retrospective analysis of the same combination in animal models and as a prospective study of this combination for future clinical trials. For purposes of comparison, parallel studies were carried out using methotrexate (MTX) and Cis-Pt. Dose-effect relationships were analyzed by the median-effect principle and the combination index-isobologram technique. EDX was the most cytotoxic agent of the three examined. The doses effective in 50% inhibition of the cell proliferation (ED50 values) for EDX, MTX, and Cis-Pt were 0.001, 0.0043, and 1.08 microM, respectively. Synergism occurred at effect levels corresponding to greater than 65% inhibition of cell growth by EDX + Cis Pt, with an increase in synergism being observed at high doses. By contrast, MTX + Cis-Pt exhibited moderate synergism, with a decrease in synergism being noted at high doses. Preceding one drug by another for 4 h during the 48-h incubation period did not result in synergism greater than that produced by simultaneous exposure to both drugs for both pairs of combinations. Due to the synergism arising from these combinations, the ED90 values can be reduced by as many as 52 and 7.3 times for Cis-Pt and EDX, respectively, as compared with only 4.0 and 1.9 times for Cis-Pt and MTX, respectively. The calculation of these drug interactions was carried out automatically with the use of computer software and was also illustrated by a sample calculation performed without computer simulation. PMID- 8422688 TI - Pharmacokinetics of peptichemio in myeloma patients: release of m-L-sarcolysin in vivo and in vitro. AB - Peptichemio (PTC) is a mixture of six synthetic oligopeptides, each of which contains the alkylating residue m-[di(2-chloroethyl)amino]-L-phenylalanine (L mSL). The fate of PTC was investigated in eight patients with multiple myeloma after intravenous infusion of the drug. The quantitative analysis of the plasma samples was performed by liquid chromatography with fluorometric detection. L-mSL was rapidly released from the peptides and reached its maximal plasma concentration at the end of the infusion. Its median elimination half-life was 1.73 (range, 0.72-2.41) h. It was possible to follow the concentration of only one of the peptides, L-mSL-L-Arg(NO2)-L-Nval.OEt, during and shortly after the infusion of PTC. The stability of L-mSL and the peptides was studied in buffer solution (pH 7.3), plasma, and blood. The stability of some of the peptides was drastically decreased in blood, the degradation half-lives being only about 1 min. We conclude that L-mSL plays an important role in the mechanism of action of PTC. PMID- 8422689 TI - Prolonged retention of high concentrations of 5-fluorouracil in human and murine tumors as compared with plasma. AB - Concentrations of 5-fluorouracil (5-FU) and its active metabolite 5-fluoro-2' deoxy-5'-monophosphate (FdUMP) were measured in biopsy specimens of tumor tissue, normal mucosa, metastatic liver nodules, and normal liver tissue obtained from 39 patients and in two murine colon tumors (colon 26 and colon 38) after a single injection of 5FU at a therapeutic dose (500 mg/m2 and 100 mg/kg, respectively). These data were compared with plasma concentrations. Peak plasma concentrations (300-500 microM) of 5FU were comparable in human and murine plasma. The half-life of plasma elimination (during the period from 15 to 120 min) in both mouse and man ranged from 10 to 20 min, whereas at between 2 and 8 h, plasma concentrations varied from 0.1 to 1 microM, the half-life being about 100 min. In both species, 5FU could be measured in plasma at concentrations ranging from 0.01 to 1 microM for several days after 5FU treatment. 5FU concentrations in tissue samples obtained from 14 patients were measured during the time range of 1-6 h, those in samples taken from 7 patients, during the interval of 19-27 h; and those in samples obtained from 18 patients, within the interval of 40-48 h after injection. 5FU tumor concentrations varied between 0.78-21.6, 0.44-6.1, and 0.17 10.8 mumol/kg wet wt., respectively. Some of the 48-h samples were obtained from patients who had received leucovorin plus 5FU; coadministration of leucovorin did not alter 5FU tissue concentrations. At between 4 and 48 h, the tissue concentration/plasma concentration ratio was at least 10. 5FU concentrations in murine tumors were measured for up to 10 days after 5FU administration, with plateau 5FU tumor concentrations being about 50 mumol/kg wet wt. in colon 38 and about 200 mumol/kg wet wt. in colon 26 at 2 h after treatment; after 4 days, values of 0.5 and 4.8 mumol/kg, respectively, were obtained and after 10 days, respective concentrations of 0.1 and 0.07 mumol/kg were detected. The FdUMP concentrations measured in colon 26 and colon 38 tumors were 214 and 46 pmol/g, respectively, at 2 h after 5FU administration, and these values subsequently decreased to about 15 pmol/g in both tumors. In human tumors the initial FdUMP concentration ranged from 10 to 1000 pmol/g; at later time points the level of FdUMP was just above the detection limit of the assay. In liver metastases, high 5FU concentrations seemed to be related to high levels of FdUMP, which was likely of importance for the antitumor effect.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8422690 TI - Relationship between the melanin content of a human melanoma cell line and its radiosensitivity and uptake of pimonidazole. AB - The intra-cellular uptake of the weakly basic radiosensitiser pimonidazole (PIMO) was determined as a function of the pigmentation of Na11+ human melanotic melanoma cells in vitro. Two experimental conditions were considered: exponentially growing cells (Exp.) and plateau-phase cells (PI.). The melanin content of Na11+ cells ranged from 500 micrograms/g cell weight in exponentially growing cells to 6000 micrograms/g in heavily pigmented plateau-phase cells. Cells were exposed to PIMO (medium dose, 0.2 mmol/dm3; 58.2 micrograms/ml). The intra-cellular concentration ranged from 163 micrograms/g in Exp. to 900 micrograms/g in pigmented Pl.; the latter being equivalent to an intra- to extracellular concentration ratio (Ci/Ce) of 17. However, this increase in the cellular uptake of PIMO was not accompanied by an increase in radiosensitising efficiency. In comparison, the Ci/Ce for etanidazole (ETA), a radiosensitiser that is uncharged at physiological pH, remained approximately constant at 1 for all values of melanin contents. Treatment of Na11+ tumours in vivo with [3H]-PIMO resulted in a tumour:blood ratio of about 3 at 30-60 min after administration. However, at 24 h a grain count of label derived from [3H]-PIMO showed that picnotic areas of tumours contained levels that were some 40 times greater than the background value. This high level of label was coincident with areas of highest apparent melanin content. In conclusion, PIMO accumulates in very heavily pigmented melanoma cells present in necrotic zones with picnosis. As these cells are probably non-clonogenic, PIMO is not suitable for use in melanoma radiotherapy. PMID- 8422691 TI - Phase I clinical and pharmacology study of 502U83 given as a 24-h continuous intravenous infusion. AB - 502U83 is an arylmethylaminopropanediol that displays significant antitumor activity in a number of murine and human tumor-model systems. In the present phase I study, a 24-h continuous intravenous infusion of this agent was given every 28 days to patients with advanced or refractory solid tumors. In all, 46 patients received a total of 96 cycles of 502U83 at doses ranging from 25 to 8,000 mg/m2. No significant hematologic, gastrointestinal, or neurologic toxicity was observed. At doses of 2,000 mg/m2 and higher, prolongation of the corrected QT interval on ECG was evident in most patients but was completely reversible, was not associated with arrhythmias, and was not dose-limiting. Dose-limiting pulmonary toxicity characterized by acute onset of dyspnea, severe hypoxemia, interstitial pulmonary edema, and death occurred in three patients treated at the highest dose levels. Plasma concentrations of 502U83 and its metabolites were measured by high-performance liquid chromatography. The 502U83 maximal concentration (Cmax) and area under the concentration-time curve (AUC) were proportional to the delivered dose; however, substantial interpatient variability in total body clearance was noted at all dose levels. Significant conversion of 502U83 to two glucuronide metabolites was detected. Metabolite concentrations were highest in the three patients who succumbed to pulmonary toxicity, although the precise contribution of these metabolites to the observed toxic effects is unknown. In view of the unfavorable clinical profile of QTc prolongation and pulmonary toxicity produced by 502U83, further clinical development of this agent has been suspended. PMID- 8422692 TI - Correlation of the in vitro cytotoxicity of ethyldeshydroxysparsomycin and cisplatin with the in vivo antitumour activity in murine L1210 leukaemia and two resistant L1210 subclones. AB - The cultured murine leukaemia L1210 cell populations used in the present study were derived from L1210 cells that had been grown in vivo. Subclones resistant to sparsomycin (L1210/Sm) or cisplatin (L1210/CDDP) were also developed in vivo. The doubling times of the cultured cell populations were identical. Fractions surviving after drug treatment in vitro were determined by colony formation in soft agar. The results, based on the differential sensitivity of the cell populations to ethyldeshydroxysparsomycin (EdSm) and CDDP, indicated that after a short exposure, cultured L1210/CDDP cells were cross-resistant to EdSm. L1210/Sm cells, however, were not cross-resistant to CDDP. The results obtained in cultured cell populations were confirmed in vivo. CD2f1 mice bearing i.p. implants of 1 x 10(5) tumour cells were given EdSm or CDDP and a combination of the two agents. Drugs were given once daily every 4 days for 3 doses starting at 24 h after tumour implantation. Treatment of mice bearing L1210/wt leukaemia with combined EdSm and CDDP caused strongly synergistic antitumour activity. In animals bearing the two resistant subclones, however, combined drug treatment did not improve the antitumour activity. The corresponding median survival of mice receiving combined drug treatment was 60 days in each group containing 6 mice bearing L1210/wt, with 4-6 cures being noted; 19 days in animals harbouring L1210/Sm, with 2 cures being recorded among 6 mice; and 11 days in mice bearing L1210/CDDP, with no cure being obtained. The results of this study indicate that the synergism resulting from combined treatment with CDDP and EdSm is a function of the cellular properties of the target tumour-cell populations and is independent of host factors. PMID- 8422693 TI - Doxorubicin and local hyperthermia in the microcirculation of skeletal muscle. AB - Doxorubicin HCl (Doxo) is an established intercalating antitumor drug. Specific side effects of Doxo primarily affect the cardiac muscle tissue to cause cardiac arrhythmias and chronic cardiomyopathies. The mechanism of action of these side effects is incompletely understood. Thus, the first objective of the present study was to test whether Doxo might have a direct effect on the microcirculation of muscular tissue. We studied large and small arterioles and large venules in the cremaster muscle of rats before and after sequential infusion of 1 (low-dose) and 10 mg/kg (high-dose) Doxo. Large arterioles showed some constriction after low Doxo doses and pronounced constriction after high Doxo doses, whereas small arterioles showed a variable response to low Doxo doses. At high Doxo doses, small arterioles dilated almost maximally (80% of the maximal response to nitroprusside). The heart rate and the diameter of large venules did not change at high Doxo doses, although the blood pressure decreased. This indicates that Doxo directly affects skeletal muscle arterioles. The second purpose of this study was to determine whether local hyperthermia would influence the microcirculation of muscular tissue such that the systemic concentration of Doxo could be reduced. In this second series of experiments, we tested whether local hyperthermia would have an effect on the skeletal muscle microvasculature and whether Doxo would change that response. Local hyperthermia alone did not alter the diameter of small arterioles or large venules, but we observed constriction of large arterioles at temperatures above 37 degrees C and during continued (60 min) hyperthermia at 40 degrees C. The low dose of Doxo did not alter these microvascular diameters at 40 degrees C. However, local hyperthermia at 40 degrees C changed the response of small arterioles to low doxo doses (no vasodilation was observed). Large arterioles continued to constrict in response to Doxo during hyperthermia. These data suggest that large arteriolar responses could be partly responsible for the toxic effect of Doxo on cardiac muscle and that local hyperthermia potentiates that response. PMID- 8422694 TI - Decreased resistance to N,N-dimethylated anthracyclines in multidrug-resistant Friend erythroleukemia cells. AB - Doxorubicin-resistant Friend erythroleukemia cells, line F4-6 ADM2R, were selected by exposure of wild-type F4-6 cells to doxorubicin concentrations of up to 1 microgram/ml. In these cells, increased expression of multidrug resistance (MDR) genes was demonstrated by Northern blot analysis. The growth-inhibitory effect of doxorubicin, daunorubicin, N,N-dimethyldoxorubicin, N,N dimethyldaunorubicin, morpholinodoxorubicin, and pyrromycin was comparatively investigated in resistant and wild-type cells. The doxorubicin-resistant F4-6 cells showed approx. 200-fold resistance to doxorubicin and about 100-fold resistance to daunorubicin with respect to the drug-sensitive counterpart. A dramatic decrease in resistance was observed for the N,N-dimethylated derivatives of doxorubicin and daunorubicin as well as for the N,N-dimethylated natural anthracycline pyrromycin and for morpholinodoxorubicin. Uptake studies using [14C]-daunorubicin and [14C]-N,N-dimethyldaunorubicin in resistant F4-6 cells showed a decreased accumulation of daunorubicin but no significant reduction in N,N-dimethyldaunorubicin accumulation as compared with the wild-type cells. Treatment with verapamil led to increased intracellular levels of daunorubicin in resistant cells, whereas an excess of N,N-dimethyldaunorubicin did not have this effect. Thus, the decreased resistance of the doxorubicin-resistant F4-6 cells to the N-alkylated anthracyclines may at least in part be due to a reduced affinity of these compounds for the efflux pump. The results indicate that the dimethylation of the amino group of the anthracycline sugar moiety and its incorporation within a morpholinyl ring may overcome MDR by similar mechanisms. PMID- 8422695 TI - Antitumor activity of the aromatase inhibitor FCE 24928 on DMBA-induced mammary tumors in ovariectomized rats treated with testosterone. AB - The antitumor activity of the irreversible aromatase inhibitor FCE 24928 (4 aminoandrosta-1,4,6-triene-3,17-dione) was studied in ovariectomized and testosterone propionate (TP)-treated rats bearing 7,12-dimethylbenzanthracene (DMBA)-induced mammary tumors. This experimental condition was used as a model of postmenopausal breast cancer. TP was given s.c. three times per week for 4 weeks at a dose of 20 mg/kg per day, a treatment that is effective in maintaining tumor growth in ovariectomized rats (89% growing tumors). FCE 24928 given s.c. twice daily 6 days/week for 4 weeks at doses of 10 and 30 mg/kg per day inhibited the tumor growth-promoting effect of TP as shown by 81% and 80% tumor-regression rates. When FCE 24928 was given at various dose levels either s.c. (3, 10, and 30 mg/kg daily) or orally (10, 30, and 100 mg/kg per day), tumor regressions were observed at all doses, amounting to 63%-93% and 63%-72%, respectively. In addition, FCE 24928 given alone (30 mg/kg daily s.c.) to ovariectomized rats did not affect ovariectomy-induced tumor regression. In conclusion, following both s.c. and oral administration, FCE 24928 was effective against DMBA-induced mammary tumors in ovariectomized TP-treated rats, a postmenopausal mammary tumor model. PMID- 8422696 TI - Organ distribution and antitumor activity of free and liposomal doxorubicin injected into the hepatic artery. AB - The plasma levels, organ distribution, and in vivo antitumor activity of free and liposomal doxorubicin injected into the hepatic artery of rats bearing W256 liver tumors were studied. The administration of liposomal doxorubicin resulted in liver-tumor and liver-parenchyma doxorubicin areas under the curve (AUCs) that were 4.7- and 3.8-fold, respectively, those obtained after the administration of free doxorubicin. Spleen and plasma AUCs were also increased by 2.8 and 2.5 times, respectively, following administration of the liposomal form. In contrast, liposomal doxorubicin did not affect heart AUCs; peak doxorubicin levels in heart tissue were three times lower in animals treated with liposomal doxorubicin. Following treatment with the liposomal form, the cumulative urinary excretion of doxorubicin at 8 h was 38 times lower. In good correlation with these findings, liposomal doxorubicin (2.35 mg/kg on day 7) was more effective than free doxorubicin against liver W256 tumors as measured by tumor-growth inhibition at 5 days after treatment (16% for liposomal doxorubicin versus -53.7% for free doxorubicin, P < 0.05) and increased life span (ILS; 108% for liposomal doxorubicin versus 27% for free doxorubicin, P < 0.05). These results demonstrate that as compared with free doxorubicin, the administration of liposomal doxorubicin into the hepatic artery results in higher drug levels in the liver tumor and enhanced antitumor activity while maintaining the cardioprotective effect of the liposome carrier as suggested by the decreased peak drug levels measured in the heart tissue. PMID- 8422697 TI - Effect of toremifene on antipyrine elimination in the isolated perfused rat liver. AB - Toremifene is a triphenylethylene antioestrogen with significant antitumor activity. It is structurally very similar to tamoxifen. Both drugs undergo extensive hepatic metabolism, and tamoxifen is known to inhibit hepatic mixed function oxidases (MFO). Using the isolated perfused rat-liver model, we investigated the effect of toremifene on the elimination of antipyrine, a standard marker of MFO activity. Perfusate consisted of 20% red cells in a modified Krebs-Henseleit buffer, and 80 ml was recirculated at 14 ml/min for 3 h. High but clinically relevant steady-state toremifene levels of 3 and 10 micrograms/ml were achieved using bolus plus constant infusion into the reservoir. Elimination of 2.5 mg antipyrine was not inhibited by steady-state toremifene, but methanol (maximal perfusate concentration, 1.29%), the vehicle used for toremifene administration, caused a statistically significant increase in the antipyrine elimination half-life (mean, 1.4 +/- 0.2 h for controls vs 2.2 +/- 0.3 h for methanol; P < 0.05, n = 4). Whereas the methanol had no apparent effect on liver viability as assessed by bile flow and perfusate back-pressure, toremifene at a steady-state concentration of 10 micrograms/ml caused a statistically significant decrease in bile flow (value at 180 min, 0.22 +/- 0.05 ml/h as compared with 0.52 +/- 0.06 ml/h in the methanol control; P < 0.05) and a statistically significant increase in perfusate back-pressure (value at 180 min, 17.5 +/- 1.8 cm vs 11.0 +/- 2.6 cm in the methanol control; P < 0.05). Therefore, toremifene used at high doses can impair liver function in the isolated perfused rat liver, but it does not have any effect on antipyrine elimination. PMID- 8422698 TI - A limited sampling method for estimation of the carboplatin area under the curve. AB - A limited sampling method for estimation of the carboplatin area under the curve (AUC) from one or two plasma concentration determination is presented. The model was conceived and developed using 43 pharmacokinetic studies in 15 patients with ovarian cancer (model data set) who received carboplatin in combination with cyclophosphamide. Linear regression analyses comparing the AUC and the drug concentration at a single time point (0.25-10 h after the end of the infusion) as calculated from the fitted exponential equations gave correlation coefficients as high as 0.97, with maximal correlations falling within the interval of 2-3.25 h. The model was validated prospectively in 9 patients with ovarian cancer (validation data set) who received the same treatment as did the model data set (21 pharmacokinetic studies), testing the equation AUC = 0.52 x C2.75 h + 0.92. Observed and estimated AUCs were correlated in the validation data set (r = 0.91). The mean predictive error (MPE% +/- SE) was -4.4% +/- 3.1% and the root mean squared error (RMSE%) was 13.9%. Multiple regression analysis revealed that adding a second sample drawn at 0.25 h (AUC = 0.053 x C0.25h + 0.401 x C2.75h + 0.628) improved the MPE% to -2.2% +/- 2.1% and the RMSE% to 9.4% (r = 0.96). We conclude that the carboplatin AUC can be estimated from a single plasma sample at 2.75 h or, more precisely, from two plasma samples at 0.25 and 2.75 h. The methods described may prove to be a handy tool for the calculation of approximate AUCs in trials of a size that would discourage detailed pharmacokinetic studies. PMID- 8422699 TI - Phase I clinical evaluation of [SP-4-3(R)]-[1,1-cyclobutanedicarboxylato(2-)](2 methyl-1,4- butanediamine-N,N1) platinum in patients with metastatic solid tumors. AB - The development of clinically useful drugs is a priority of clinical cancer research. CI-973, [SP-4-3(R)]-[1,1-cyclobutanedicarboxylato(2-)](2-methyl-1,4- butanediamine-N,N1) platinum, has been shown in preclinical murine and human tumor models to have activity equivalent or superior to that of cisplatin and carboplatin and to exert activity against cisplatin-resistant cell lines. In addition, preclinical testing suggests a reduced toxicity profile for CI-973 as compared with currently available drugs, especially decreased nephrotoxicity, ototoxicity, and gastrointestinal toxicity. A total of 29 (28 evaluable) patients with solid tumors were treated with intravenous CI-973 given over 30 min every 4 weeks. No routine pre- or post-treatment hydration or antiemetic program was used. The CI-973 doses given were 75, 150, 170, 188, 230, and 290 mg/m2. The dose limiting toxicity was granulocytopenia. Nausea and vomiting occurred in the majority of patients but was mild to moderate in severity. No renal or auditory toxicity was seen. The maximum tolerated dose (MTD) for patients who had a good performance status, had not received prior radiation therapy to bone marrow, and had not previously been exposed to platinum or stem-cell toxin was 290 mg/m2. For those who had received prior radiation therapy, had a performance status of 2 or worse, or had previously been exposed to platinum or stem-cell toxin, the MTD was 230 mg/m2. The recommended phase II starting doses for these groups of patients are 230 and 190 mg/m2, respectively. No clinical tumor response was seen in this phase I study. PMID- 8422700 TI - Ifosfamide in advanced epidermoid head and neck cancer. AB - A total of 37 men with epidermoid head and neck cancer whose disease had recurred following primary treatment (surgery and/or radiotherapy) received first-line chemotherapy with ifosfamide at i.v. doses of 3 g/m2 given daily on 3 consecutive days in combination with mesna (600 mg/m2 x 3 oral daily doses on days 1-3) every 3 weeks. In all, 7 patients showed a partial response and 2 patients achieved a complete response, for an overall objective response rate of 26% (9 of 35 eligible patients; 95% confidence interval, 12.5%-43%). Excluding the 5 early nontoxic deaths observed during the first 3 weeks of therapy, the objective response rate was 30% (9 of 30 patients; 95% confidence interval, 15%-49.5%). Responses were seen in lung metastases (2 patients), lymph nodes (2 patients), skin (3 patients), and cases of local recurrence (5 patients). The median duration of responses was 3 months (range, 2-5 months). The main side effects of ifosfamide were alopecia (83% of patients), emesis (80%), granulocytopenia (23%), and mild mucositis (20%). Two poor-risk patients suffered severe CNS complications that were probably related to treatment. Three patients died due to chemotherapy-related complications (2 patients with CNS toxicity and 1 patient with granulocytopenic sepsis). In conclusion, ifosfamide appears to be an active drug in epidermoid head and neck cancer and merits further evaluation in this disease. PMID- 8422701 TI - Tumor progression and metastasis in murine D2 hyperplastic alveolar nodule mammary tumor cell lines. AB - We have examined tumor progression and metastatic properties of three clonal murine mammary tumor cell lines of recent origin (D2A1, D2.OR and D2.1). These lines were derived from spontaneous mammary tumors which originated from a D2 hyperplastic alveolar nodule (HAN) line. D2A1 cells were more malignant than D2.OR or D2.1 cells, whether measured by experimental metastasis assays after intravenous injection in nude mice or chick embryos, in vivo growth rate of primary tumors following mammary fat pad injection in nude mice, or spontaneous metastasis assay from primary tumors growing in mammary fat pads. D2A1 cells also were more invasive in vitro in a Matrigel invasion assay than D2.1 cells, while the D2.OR cells were non-invasive in this assay. The increased invasiveness and malignancy of D2A1 cells were associated with increased levels of mRNA for the cysteine proteinase cathepsin L. Levels of osteopontin (OPN), nm23, int-1 and int 2 mRNAs were also examined. Nm23 levels were highest in the most malignant cell line. These cell lines provide a model for studying the tumorigenic and metastatic ability of mammary tumor cells and offer several advantages: they were cloned from mammary tumors that originate from a common source of preneoplastic cells (D2HAN); they are of relatively recent origin; and they have spontaneously arrived at different stages of tumor progression. PMID- 8422703 TI - Expression of ganglioside GM3 and H-2 antigens in clones with different metastatic and growth potentials isolated from Lewis lung carcinoma (3LL) cell line. AB - In view of the evidence that cell expression of gangliosides in several tumors is positively involved in the metastatic phenotype, Lewis lung carcinoma (3LL) cell line, expressing GM3 as the major ganglioside, was analysed for the cell surface expression of GM3. An indirect immunofluorescence assay, using a M2590 monoclonal antibody recognizing GM3, was used for this purpose. Since the parental 3LL cells consist of heterogenous subpopulations differing in the degrees of GM3 expression, we have developed clones of this cell line with different degrees of metastatic potentials by using an in vitro non-selective procedure in order to investigate whether the expression of GM3 is associated with metastatic potential. The degree of cell surface expression of GM3 among the clones correlated well with their total cellular content of this ganglioside. However, we were unable to confirm the report of increased level of GM3 in high metastatic 3LL clones, nor did a decreased level correlate with weak metastatic ability. In our recent work, an inhibitor of glucosylceramide synthase, D-threo-1-phenyl-2 decanoylamino-3-morpholino-1-propanol (D-PDMP), was found to decrease the levels of all cellular glucosphingolipids and cause the accumulation of the precursors of glucosylceramide. The present study does not, however, rule out the possible involvement of this lipid family in metastatic dissemination, since treatment of 3LL cells with D-PDMP resulted in significant inhibition of their experimental metastatic potential. Clones expressing very low GM3 grew slowly in culture dishes, suggesting that GM3 may have a regulatory role in cell proliferation. The low metastatic clones expressed high levels of H-2Kb antigen, while the expression of the same antigen on the high metastatic clones was relatively low, confirming the previous observation of this tumor system. Moreover, a clone showing the lowest tumorigenic potency revealed both a high cell surface expression of H-2Kb and a high H-2Kb/H-2Db ratio. PMID- 8422702 TI - Rapid effects of EGF on cytoskeletal structures and adhesive properties of highly metastatic rat mammary adenocarcinoma cells. AB - In the highly metastatic rat mammary adenocarcinoma cell clone MTLn3, EGF induced increased adhesion to fibronectin while in the human epidermoid carcinoma cell line A431 EGF induced diminished adhesive properties. Flattening of cells with extensive formation of filopodia was observed in MTLn3 cells within 5 min of EGF addition, while in A431 cells EGF induced rounding up and only occasional formation of filopodia. Immunofluorescent analysis revealed extension of microtubules (MT) into the filopodia and Western blot analysis demonstrated an EGF-induced 2- to 3-fold increase in the amount of assembled tubulin in MTLn3 but not in A431 cells. In MTLn3, but only marginally in A431 cells, EGF treatment resulted in phosphorylation of a 280 kD cytoskeleton-associated protein, which was rapid and dose-dependent. These results suggest differential signal transduction pathway of cytoskeleton-associated EGFRs in highly metastatic MTLn3 as compared with A431 cells. PMID- 8422704 TI - Expression of annexins on the surfaces of non-metastatic and metastatic human and rodent tumor cells. AB - Annexins are a large group of calcium-dependent cytoskeletal- and membrane associated proteins whose properties include cytoskeleton and phospholipid binding and mitotic signal transduction. Although annexin-like molecules have been reported on the external plasma membranes of certain cells, in general they are considered to be cytoplasmic proteins. We report here the heterogenous expression of certain annexins (I-VI) on the external cell surfaces of non metastatic and metastatic murine (RAW117 large-cell lymphoma), rat (13762NF mammary adenocarcinoma) and some human (KM12 and HT29 colorectal carcinoma) cell lines but not on some other cell lines such as human (A375 and MeWo) and mouse (B16) melanoma. The implication of annexin cell surface expression in the metastatic process is discussed with respect to tumor cell adhesion. PMID- 8422705 TI - A spontaneous subcutaneous tumor in C57BL/6 mice that metastasizes to the liver. AB - A malignant tumor that arose spontaneously in the subcutaneous tissue of the back of a C57BL/6 female mouse was found to metastasize spontaneously to the liver. The primary and metastatic tumors, SML (spontaneous metastasis to the liver) 1 and SML 2, were established in vitro in long-term cell suspension culture and were passaged 10 times in vivo for 18 months. When 100,000 cells were injected subcutaneously in the orthotopic position, tumor growth appeared in 60% of the SML 1 mice and 100% of the SML 2 mice. SML 1 did not grow when injected in the footpad, while SML 2 did. The median survival was 47 days for SML 1 and 48.5 days for SML 2 (P = 0.013). The pattern of metastasis was similar for both tumor cell lines, irrespective of intravenous or subcutaneous injection routes. Spontaneous metastasis of the SML 2 line occurred from both the orthotopic and heterotopic sites, while the SML 1 metastasized spontaneously from the orthotopic site only. Liver metastasis appeared in > 90% of the mice for both SML 1 and SML 2. Metastasis to the spleen occurred in about half the mice. Other sites of metastasis were the ovaries (36% and 52%, respectively, for SML 1 and SML 2), the kidneys (approximately 15%) and the small bowel (very rarely). Metastasis to the lungs did not occur except very rarely in the later passages of the SML 2 line. Histologic, immunohistochemical and electron microscopic studies showed a histiocytic tumor with macrophage characteristics. The cells exhibited chemotaxis toward liver extracellular matrix and reduced motility toward collagen IV, laminin and fibronectin compared to the B16-F10 melanoma line. This spontaneously occurring tumor should prove useful for the study of organ-specific metastasis to the liver. PMID- 8422706 TI - Importance of orthotopic implantation for human tumors as model systems: relevance to metastasis and invasion. AB - Transplantation of human tumors into immunodeficient athymic nude mice has become an important experimental approach to study the biology and the treatment of human cancer. Most human tumor xenograft experiments have employed subcutaneous injection procedures, but the main limit of this technique is the lack of metastasis from the subcutaneous site. The possibility of producing experimental metastasis by intravenous injection of cells in the animals has been known for a long time, and it has been recently reported that tumorigenic properties and metastatic ability of human cancer can be altered by transplantation of the tumor into its organ or tissue of origin in the recipient animals (orthotopic transplantation). In this paper we review (1) the principal techniques of orthotopic injection of most solid tumors, (2) the most recent techniques to achieve experimental metastases, and (3) the methods for preparing tumor cell suspensions from human surgical specimens suitable for transplantation into animals. These animal models should be used for a more appropriate evaluation of new antitumor treatments including the ones targeted to inhibit metastatic spread. PMID- 8422707 TI - Modulation of invasive potential in different clonal subpopulations of a rat rhabdomyosarcoma cell line (BA-HAN-1) by differentiation induction. AB - Three clonal subpopulations (A, B, C) isolated from the same rhabdomyosarcoma of the rat and differing in their degree of spontaneous differentiation were tested for their invasive potential before and after differentiation induction with retinoic acid (RA), N-monomethylformamide (NMF) and sodium butyrate (NaBut). Invasive potential was analysed in an in vitro assay using embryonic chick heart fragments in organotypic culture. In standard culture medium, all three subpopulations were shown to be invasive, progressively replacing the chick heart fragments within 7-11 days after confrontation. After exposure to RA, NMF or NaBut, marked differences in the invasive potential of these subpopulations were, however, observed. Subpopulation C exhibited a pronounced decline in invasive potential, as evidenced by a significant decrease (P = 0.005) in the proportion of chick heart fragments with advanced stages of invasion. This response, however, was confined to the differentiation-inducing agents RA and NaBut, which had also produced a marked increase in morphological and/or biochemical differentiation (P = 0.0001). In contrast, NMF, which had only minor effects on differentiation, failed to affect the invasive potential of subpopulation C. In subpopulation B, a transient inhibition of single cell invasion became evident after exposure to RA, whereas NMF and NaBut failed to affect the invasive potential in spite of minor effects on differentiation. In the least differentiated subpopulation A, which was shown to be refractory to the differentiation-inducing effects of RA, NMF and NaBut, there was also no observation of any reduction of invasive potential. The results of our study demonstrate that differentiation-inducing agents can significantly reduce the invasive potential of malignant tumors, although marked differences of response are to be expected between the different subpopulations of a tumor. PMID- 8422708 TI - Effects of metastatic and non-metastatic mammary adenocarcinoma on tissue distribution of gallium-67 in the rat. AB - The purpose of this work was to study the effect of metastatic and non-metastatic mammary adenocarcinoma on tissue distribution of gallium-67 (67 Ga) citrate in Fischer-344 female rats by the use of gamma counting techniques. The homogenate (0.1 mm) of a sample of metastatic and non-metastatic tumor was implanted by subcutaneous injection in the right footpad of each animal's hind extremity. The animals bearing metastatic tumor were studied 2-24 days and the non-metastatic group 2-30 days after the implantation of tumor homogenate. The control group consisted of four animals and tumor-bearing groups of seven to eight animals at each time point. All animals were injected with 1.11 MBq of 67Ga citrate by intravenous administration and sacrificed in halothane anesthesia 48 h later. The relative tissue uptake data are presented as arithmetical mean value with a standard error and graphically demonstrated as normalized data with respect to control. The results demonstrate that 67Ga citrate uptake was largely unaffected in most organs by the presence of either metastatic or non-metastatic tumor. Gallium-67 uptake, however, was significantly and consistently increased in the popliteal lymph nodes of the ipsilateral extremity of tumor implant in the metastatic group. No difference was observed in the non-metastatic tumor group. The findings of this experimental work indicate that the host reaction to the tumor does not modify the gallium uptake characteristics in the normal tissues of tumor-bearing animals. PMID- 8422709 TI - Repression of stromelysin metalloprotease expression in rat fibrosarcoma cells by dimethylsulfoxide. AB - Metalloproteases are implicated in conferring invasive properties to tumor cells. We show here that treatment of ras-oncogene-transformed rat fibroblasts with dimethylsulfoxide (DMSO) results in a reversible decrease in stromelysin mRNA. Furthermore, stromelysin expression was found to be repressed by DMSO, but not by glucocorticoid hormone, in a fibrosarcoma cell line showing low AP-1 (fos/jun) transcription factor activity. In two fibrosarcoma cell lines which express high levels of stromelysin and low levels of 68 kDa type IV collagenase, the DMSO induced decrease in stromelysin expression was paralleled by a decreased invasive propensity. PMID- 8422711 TI - Scintigraphic detection of bile leaks after laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy is widely accepted as an alternative to traditional open cholecystectomy. Its advantages are a shorter hospital stay, earlier return to normal activity, less pain, and improved cosmetic results. However, in the initial experience with this technique, biliary leaks were several times more frequent than in traditional cholecystectomy. Four cases of scintigraphic detection of biliary leaks after laparoscopic cholecystectomy are described. The results of the examination are compared in two cases with endoscopic retrograde cholangiography and in the remaining two with intraoperative findings. Hepatobiliary scintigraphy is suggested for early evaluation of postoperative complications involving the biliary system. PMID- 8422712 TI - Septic pulmonary emboli from pulmonic valvular endocarditis demonstrated by serial ventilation-perfusion lung imaging. AB - The evolution of multiple ventilation-perfusion mismatches on serial lung scintigraphy in a fully anticoagulated, bacteremic patient with small pulmonary infiltrates and repeatedly negative Doppler ultrasonography of the lower extremities suggested pulmonary emboli from right-sided valvular endocarditis. The demonstration of pulmonic valvular vegetations on follow-up transesophageal echocardiography supported this diagnosis. In the proper clinical setting, septic pulmonary emboli should be included in the differential diagnosis of high probability ventilation-perfusion lung imaging. PMID- 8422710 TI - Adriamycin-induced inhibition of melanoma cell invasion is correlated with decreases in tumor cell motility and increases in focal contact formation. AB - Tumor cell adhesion to the extracellular matrix (ECM) is closely linked with tumor cell invasion and metastasis. In this study, we demonstrate that low levels of adriamycin, a widely used anticancer drug, can inhibit the invasion of highly metastatic K1735-M2 mouse melanoma cells in vitro through a reconstituted basement membrane extract. Adriamycin-induced inhibition of melanoma cell invasion occurred at levels of the drug (i.e. 1 ng/ml) that did not inhibit tumor cell growth, suggesting that the observed inhibition in tumor cell invasion was not due to the well-documented ability of adriamycin to interfere with DNA and/or RNA synthesis. Rather, these studies indicated that adriamycin-induced inhibition of melanoma cell invasion was accompanied by a corresponding decrease in the ability of adriamycin-treated tumor cells to migrate in response to several isolated ECM components including fibronectin, laminin and basement membrane (type IV) collagen. The decreased migration of adriamycin-treated tumor cells was not accompanied by a decrease in the adhesion or spreading of the adriamycin treated cells on substrata coated with these ECM components. Instead, adriamycin treated cells actually exhibited a slightly increased propensity (compared to untreated control cells) to adhere on fibronectin-, laminin-, and type IV collagen-coated substrata. Additionally, adriamycin treatment caused a dramatic increase in focal contact formation by these melanoma cells, as assessed by fluorescent microscopy of actin and vinculin. In addition to providing a useful model for which to study the molecular and cellular basis for focal contact formation, these results further emphasize the results of several other investigators that have suggested an important role for focal contacts in modulating tumor cell motility, invasion and metastasis. PMID- 8422713 TI - Comparison of Tc-99m DTPA aerosol ventilation studies with pulmonary function testing in cystic fibrosis. AB - In 43 patients with cystic fibrosis, the results of 122 Tc-99m DTPA aerosol ventilation (DAV) studies were compared with pulmonary function tests (PFTs) that were performed within 24 hours of the DAV studies. The DAV studies were evaluated blindly for (A) number of pulmonary segments showing little or no ventilation, (B) number of foci of bronchial deposition of aerosol, and (C) subjective overall improvement, lack of change, or worsening from the previous study. (A) and (B) correlated significantly with all PFTs (p's < .001, r's = -.51 to -.73). Changes in (A) and (B) also correlated with changes in PFTs (p's < or = .001, r's = -.37 to -.58). The three populations in (C) were significantly different from each other with respect to changes in all PFTs (p < or = .002). Intervals between studies showing subjective improvement, no change, and worsening averaged 60, 133, and 306 days, respectively. These results suggest that DAV is an indicator of both regional and global pulmonary function and may be useful in evaluating patients with cystic fibrosis. PMID- 8422714 TI - Respiratory distress syndrome. A suggestive pattern of shunt effect detected by means of macroparticles. AB - An esophagobronchial fistula developed in a patient who had well-differentiated squamous carcinoma of the lung that was treated with chemotherapy. Because the esophagobronchial fistula could not be surgically repaired, it was isolated with a mechanical stitch above and below it. Forty-eight hours after initiation of enteral nutrition, a perfusion lung scan was performed because of clinical suspicion of pulmonary embolism. Because the scan showed reduced pulmonary radioactivity and accumulation of activity in the kidneys and spine, an arteriovenous shunt was suspected. However, subsequent digital subtraction angiography ruled out this possibility and a recurrence of the esophagobronchial fistula was confirmed with an esophagogram. The unusual extrapulmonary activity could be related to a reversible capillary shunt in the pulmonary vasculature, secondary to acute respiratory distress syndrome. PMID- 8422715 TI - Tc-99m MAG3 as an alternative to Tc-99m DTPA and I-131 Hippuran for renal transplant evaluation. AB - This study investigated the new radiopharmaceutical, Tc-99m mercaptoacetyltriglycine (MAG3), as a possible alternative to Tc-99m DTPA and I 131 Hippuran for renal transplant evaluation. Tc-99m MAG3 and I-131 Hippuran were used sequentially in 19 consecutive patient studies and compared qualitatively and quantitatively. Serum creatinine values ranged from 1.6 mg/dL to 10.3 mg/dL (mean, 5.4 mg/dL). The Tc-99m MAG3 images and time-activity curves were of superior quality (3+) compared to I-131 Hippuran (1-2+). Quantitative analysis of the time to peak activity (Tmax) demonstrated a high correlation and no significant difference for Tc-99m MAG3 (mean, 11.4 min) compared to I-131 Hippuran (mean, 11.2 min). The percentage of peak activity retained at 20 minutes (T20) also showed a high correlation between the two agents; however, Tc-MAG3 showed a significantly slower clearance (T20 mean, 77%) than I-131 Hippuran (mean, 71%). Flow images were judged to be of good quality; however, the first pass time-activity curves were often different from that seen with Tc-99m DTPA. Interpretation of delayed Tc-99m MAG3 images, for example to diagnose a slow urinary leak, were sometimes complicated by bowel clearance. In conclusion, Tc 99m MAG3 is superior to I-131 Hippuran for evaluation of renal transplant function. Tc-99m is the agent of choice for evaluation of renal transplant blood flow and function. PMID- 8422716 TI - Gallbladder activity appearing 6 minutes after the intravenous injection of Tc99m MAG3 simulating a picture of obstructive uropathy of the right kidney. AB - Two cases of Tc-99m MAG3 renograms performed on the same day are presented. Both of them show considerable liver activity, which is a known drawback for MAG3 by distribution. In the first case, gallbladder activity appeared after 6 minutes, at the same time when tubular drainage and caliceal activity started to show in both kidneys. The gallbladder activity was superimposed over the upper calyces of the right kidney and simulated an obstructive pattern in the time-activity curves. This early gallbladder activity after intravenous injection of Tc-99m MAG3 has not been previously reported. The second case is presented for the purpose of confirming that nothing in the preparation of the kit contributed to this phenomenon. PMID- 8422717 TI - Clinical and prognostic effect of a positive granulocyte scan in infective endocarditis. AB - Thirty patients with clinical signs of infective endocarditis and pathologic echocardiographic findings indicating vegetations underwent scanning with In-111 or Tc-99m hexamethylpropylene amineoxime (HMPAO) labeled granulocytes. Blood cultures were positive in 60% of the patients. The other cases were negative as a result of antibiotic pretreatment. The results of scintigraphy were correlated with the subsequent clinical course, and in 20 cases with data obtained by histologic examination of the valves. With regard to the degree of the inflammatory process, the nuclear medical procedure provided the following results: true-positive in 6 cases, false-positive in 1 case, true-negative in 19 cases, and false-negative in 3 cases. In this study, positive granulocyte scans correlate with high activity of the inflammatory process and predict a poor prognosis for the patients concerned. PMID- 8422718 TI - False-positive Tc-99m sestamibi SPECT in a patient with left bundle branch block. AB - A 37-year-old man with atypical chest pain and complete left bundle branch block showed a marked exercise induced septal defect on Tc-99m sestamibi stress testing. A repeat examination at rest the next day revealed complete reversal of the previous septal defect. Coronary angiography two days later showed a normal coronary artery system. Left bundle branch block has been associated with false positive results of exercise (and more recently, dipyridamole) TI-201 examinations for septal ischemia. The case presented here may be the first reported example of a false positive Tc-99m sestamibi examination for septal ischemia in the presence of left bundle block. PMID- 8422719 TI - Serial SPECT imaging in moyamoya using I-123 IMP. A method of noninvasive evaluation and follow-up. AB - Serial cerebral perfusion studies were performed in a patient with moyamoya disease, utilizing N-isopropyl iodoamphetamine (I-123 IMP). The SPECT findings correlated closely with those of a CT scan and magnetic resonance imaging (MRI). In fact, the perfusion defects seen on SPECT studies were larger than those seen on CT and MRI studies. The SPECT findings paralleled the patient's improving clinical course. Cerebral perfusion SPECT studies may be very helpful in the evaluation and follow-up of patients with moyamoya disease. PMID- 8422720 TI - High resolution Tc-99m HMPAO SPECT in a patient with transient global amnesia. AB - This is a report of a patient who underwent high-resolution brain perfusion SPECT studies during and after an episode of transient global amnesia. During the attack, SPECT imaging showed increased perfusion in the left medial temporal region involving the amygdala and hippocampus. After recovery from amnesia, a follow-up SPECT examination showed slightly decreased perfusion in this region. These findings support the hypothesis that transient global amnesia is associated with transient hyperperfusion in the medial temporal brain structures, and confirm the utility of high-resolution SPECT imaging for the evaluation of the fine details of functional brain anatomy. PMID- 8422721 TI - The effects of danazol on a patient with familial dysalbuminemic hyperthyroxinemia. AB - Familial dysalbuminemic hyperthyroxinemia, a syndrome involving an abnormal affinity of albumin for thyroxine, results in elevated total thyroxine and free thyroxine index levels but normal triiodothyronine resin uptake and thyroid stimulating hormone levels. Danazol is a synthetic androgen that increases triiodothyronine resin uptake and decreases total thyroxine, secondary to a decrease in thyroid-binding globulin levels. A 35-year-old woman with familial dysalbuminemic hyperthyroxinemia who was given danazol, in whom nervousness, insomnia, and weight gain developed, is described. Thyroid tests performed after initiation of danazol therapy revealed an increase in triiodothyronine resin uptake, with persistently elevated total thyroxine and free thyroxine index levels, and normal thyroid-stimulating hormone levels. Once the danazol was withdrawn, the symptoms resolved, the triiodothyronine resin uptake returned to normal, and the thyroid-stimulating hormone remained normal. The effects of danazol on a patient with familial dysalbuminemic hyperthyroxinemia correlate well with the effects on normals, and the ultrasensitive thyroid-stimulating hormone was the most useful test in separating hyperthyroxinemia from hyperthyroidism. PMID- 8422722 TI - Study of the tumor marker carbohydrate antigen 50 in liver cirrhosis. Pathogenetic considerations. AB - Carbohydrate antigen 50 (CA 50) is a tumor marker that increases in many malignancies, especially in carcinoma of the digestive tract. False-positive results occur in benign liver disease. The behavior of CA 50 in 86 cirrhotic patients was studied, with thorough clinical and laboratory evaluations. There were abnormal values in 75.6% of the patients without significant differences among the different Child's grades. Significant correlations with some liver tests were found, especially transaminases, and at lower degrees with cholestatic parameters. Despite the previously reported relation with cholestasis, especially in biliary diseases, the CA 50 serum levels of the authors' cirrhotic patients appeared to be more closely related to cytolysis, according to the results of several statistical tests, including multivariate analysis. Because of the percentage and the levels of the abnormal results, this antigen cannot be used as a tumor marker in cirrhotic patients. Cytolysis seems to have a pathogenetic role in the increase of CA 50, at least in cirrhosis. PMID- 8422723 TI - Bladder variants noted on bone and renal imaging. PMID- 8422724 TI - Scintigraphic detection of transient afferent loop obstruction. PMID- 8422725 TI - Pulmonary embolism from a popliteal vein aneurysm. AB - This is the first report of a venous aneurysm detected by radionuclide venography. Aneurysms of the popliteal vein are very rare. Described here is a case of a saccular popliteal venous aneurysm that presented as dyspnea secondary to pulmonary embolism. In this case, there was recurrent PE despite anticoagulant therapy. Almost all such aneurysms are associated with PE, about half of which are recurrent. This report demonstrates another useful aspect of obtaining a simultaneous radionuclide venogram when performing a perfusion lung scan with Tc 99m MAA. PMID- 8422726 TI - Skeletal metastasis of malignant glucagonoma mimicking avascular necrosis of the hip scintigraphic and MRI correlation. PMID- 8422727 TI - Unilateral medullary sponge kidney detected on bone scan. PMID- 8422728 TI - Labeled leukocyte imaging in chemical cystitis. PMID- 8422729 TI - Blood pool imaging of portosystemic collaterals. PMID- 8422730 TI - An active bleeding gastric ulcer demonstrated by Tc-99m RBC gastrointestinal bleeding study. PMID- 8422731 TI - Cardiac blood pool visualization by Ga-67 scintigraphy at 48 hours in a patient with severe heart failure. PMID- 8422732 TI - Radiation effects on pulmonary ventilation and perfusion. PMID- 8422733 TI - High spatial resolution Tc-99m HMPAO brain SPECT in cerebellar embolic infarction. PMID- 8422734 TI - Three-dimensional surface display with I-123 IMP of slowly progressive apraxia. PMID- 8422735 TI - Mall de Pott. A cause of photopenic lesion on Tc-99m HMPAO-labeled leukocyte scintigraphy. PMID- 8422737 TI - Valproic acid pharmacokinetics in children. IV. Effects of age and antiepileptic drugs on protein binding and intrinsic clearance. AB - Pharmacokinetic data from 48 children who were taking valproic acid were analyzed by multiple stepwise linear regression. Children who were receiving enzyme inducing antiepileptic drugs (n = 27) had greater (p < 0.01) clearances, elimination rates, and dosage requirements and greater (p < 0.05) variability in pharmacokinetic values than patients receiving monotherapy. Age and polytherapy explained most of the interpatient variability in total (r2 = 0.80; p < 0.001) and intrinsic (r2 = 0.77; p < 0.001) clearances and the elimination rate (r2 = 0.61; p < 0.002). Free fraction variability was related to valproate concentration and phenobarbital (r2 = 0.47; p < 0.001). Distribution volume variance was associated with free fraction (r2 = 0.48; p < 0.001). The effect of age and polytherapy on valproate clearance is primarily attributable to changes in metabolism rather than in protein binding. Valproic acid dosage requirements are greater and more variable for children who are receiving other enzyme inducing antiepileptic drugs. PMID- 8422736 TI - The South to South Cooperation in Reproductive Health. PMID- 8422738 TI - Awakenings: using a popular movie to teach clinical pharmacology. PMID- 8422739 TI - A predictor for side effects in patients with Alzheimer's disease treated with deferoxamine mesylate. AB - In a previously reported clinical trial, patients with Alzheimer's disease were treated with deferoxamine mesylate, which resulted in a 50% reduction in the average rate of deterioration over 2 years. There were five deaths in the untreated group during the trial and no deaths in the treated group, although five of 25 treated patients reported anorexia. Deferoxamine metabolite analysis of urine for 24 hours after deferoxamine injection from sensitive and nonsensitive patients showed marked differences. Occurrence of side effects correlated with increased formation of a monoamine oxidase catalyzed (major) metabolite, MFO1. The metabolite ratio, MFO1/total metabolites, plus parent drug (TOT) showed a bimodal distribution with a mean +/- SD value of 0.68 +/- 0.06 for the nonsensitive and 0.79 +/- 0.04 for sensitive patients. The MFO1/TOT ratio discriminates between sensitive and nonsensitive patients, and we suggest that the half difference mark between the two mean values (0.735) can be used as a predictor of side effects. Patients with a MFO1/TOT ratio of greater than 0.70 would be considered at risk and observed for onset of side effects. Patients with a MFO1/TOT ratio greater than 0.80 would be considered for immediate adjunct treatment with isoniazid or other monoamine oxidase inhibitors. PMID- 8422740 TI - The pharmacokinetic and pharmacodynamic interaction between propafenone and lidocaine. AB - Although propafenone is a known substrate and inhibitor of the cytochrome P450 4 hydroxylation pathway of debrisoquin (CYP2D6 isozyme), its effects on other hepatic mixed- function oxidative isozymes have not been extensively evaluated. We studied the influence of propafenone on the disposition of continuously infused lidocaine in 12 healthy male volunteers. Placebo or propafenone (225 mg every 8 hours) was orally administered for 4 days before and during lidocaine administration (2 mg/kg/hr for 22 hours). In the 11 (92%) subjects phenotyped as extensive metabolizers, propafenone significantly increased the lidocaine area under the plasma concentration time curve (81.7 +/- 16.2 versus 76.3 +/- 15.6 micrograms.hr/ml; p < or = 0.05) and reduced systemic lidocaine clearance (9.53 +/- 1.77 versus 10.27 +/- 2.24 ml/min/kg; p < or = 0.05), but did not significantly affect volume of distribution at steady state (2.48 +/- 0.33 versus 2.64 +/- 0.45 L/kg; p = 0.10) or mean residence time (4.37 +/- 0.92 versus 4.47 +/- 0.87 hours; difference not significant) compared with placebo, respectively. Adverse central nervous system effects were significantly worse in severity and duration during the propafenone phase (p < or = 0.05). Propafenone minimally inhibits the metabolism of lidocaine. This suggests that the ability of propafenone to inhibit metabolic pathways exclusive of the CYP2D6 isozyme may be limited. In addition, potentiation of disturbing central nervous system adverse effects may occur during combination therapy of propafenone and lidocaine. PMID- 8422741 TI - Characterization of glutathione conjugation in humans: stereoselectivity in plasma elimination pharmacokinetics and urinary excretion of (R)- and (S)-2 bromoisovalerylurea in healthy volunteers. AB - Characterization of glutathione conjugation in vivo was performed in 12 healthy male volunteers by use of the racemic drug bromisovalum (bromisoval; 2 bromoisovalerylurea) as a model substrate. To study whether the pharmacokinetics of both bromisovalum enantiomers was related to the glutathione S-transferase class Mu phenotype, six subjects who were class Mu deficient and six subjects who were not class Mu deficient participated. After oral administration of 600 mg racemic bromisovalum, enantioselective measurement of unchanged bromisovalum (plasma and saliva) and the diastereomeric bromisovalum mercapturates (urine) showed a pronounced stereoselectivity in all subjects. The plasma clearance of R bromisovalum was about 12 times higher than that of S-bromisovalum (9.3 +/- 3.7 and 0.78 +/- 0.38 L/min, respectively), which was in agreement with the higher urinary cumulative excretion for the mercapturate derived from R-bromisovalum: 26% +/- 4% of the dose versus 8% +/- 3% of the dose for the mercapturate derived from S-bromisovalum. Both the bromisovalum pharmacokinetics in general and the stereoselectivity in bromisovalum pharmacokinetics were not different for the subjects who were glutathione S-transferase class Mu deficient and the subjects who were not glutathione transferase class Mu deficient. PMID- 8422742 TI - Pharmacodynamic variability of flecainide assessed by QRS changes. AB - The effect of flecainide on the QRS interval was studied in 10 patients who were receiving long-term oral treatment (50 to 150 mg twice daily) for arrhythmias that were refractory to other drugs. Total and free drug plasma levels and QRS durations were measured at intervals after the morning administration. Free drug plasma levels were linearly correlated with QRS duration in each patient and the slope of the line was widely variable in the population studied. Even after the data from one patient with an unusually high slope (0.454) was excluded from the analysis, the slope range was 0.0284 to 0.144. Pharmacodynamic variability could not be explained by heart rate changes, active metabolites, electrolyte disturbances, or free drug concentration. None of the pharmacokinetic parameters measured (average steady-state concentration, fluctuation of maximum and minimum concentrations, time to peak concentration, final half-life, and protein binding) showed an intersubject variability greater than 4.4 times. Our findings suggest that the determination of flecainide free plasma concentration may not be sufficient to forecast electrophysiologic effects in individual patients. PMID- 8422743 TI - Pharmacokinetic-pharmacodynamic modeling of caffeine: tolerance to pressor effects. AB - We propose a parametric pharmacokinetic-pharmacodynamic model for caffeine that quantifies the development of tolerance to the pressor effect of the drug and characterizes the mean behavior and inter-individual variation of both pharmacokinetics and pressor effect. Our study in a small group of subjects indicates that acute tolerance develops to the pressor effect of caffeine and that both the pressor effect and tolerance occur after some time delay relative to changes in plasma caffeine concentration. The half-life of equilibration of effect with plasma caffeine concentration is about 20 minutes. The half-life of development and regression of tolerance is estimated to be about 1 hour, and the model suggests that tolerance, at its fullest, causes more than a 90% reduction of initial (nontolerant) effect. Whereas tolerance to the pressor effect of caffeine develops in habitual coffee drinkers, the pressor response is regained after relatively brief periods of abstinence. Because of the rapid development and regression of tolerance, the pressor response to caffeine depends on how much caffeine is consumed, the schedule of consumption, and the elimination half-life of caffeine. PMID- 8422745 TI - Pharmacodynamic effects of inhaled dry powder formulations of fenoterol and colforsin in asthma. AB - The airway and tremor response and cardiovascular and hypokalemic effects of single doses of inhalative fenoterol dry powder capsules (0.4 mg) were compared with the fenoterol metered dose inhaler (0.4 mg) and colforsin (forskolin) dry powder capsules (10.0 mg), a direct activator of the adenylate cyclase system, in 16 patients with asthma. Subjects (FEV1 < or = 60% predicted) were investigated in a randomized, double-masked, placebo-controlled, four-period, crossover trial for a 120 minute period. All active drugs caused a significant increase in specific airway conductance (p < 0.05); the order of potency (mean +/- SEM maximum increase from baseline) was fenoterol metered dose inhaler (0.51 +/- 0.06 sec-1 x kPa-1), fenoterol dry powder capsules (0.49 +/- 0.07), and colforsin dry powder capsules (0.30 +/- 0.03). A marked increase in finger tremor amplitude resulted after fenoterol metered dose inhaler only (62.93% +/- 10.21%; p < 0.05) in contrast to fenoterol dry powder capsules (15.84% +/- 4.35%; p < 0.05) and colforsin dry powder capsules (12.87% +/- 10.44%; p > 0.05). A decrease in plasma potassium was found after fenoterol (metered dose inhaler > dry powder capsules; p < 0.05). In conclusion, fenoterol dry powder capsules caused less tremor response and hypokalemic effects than the metered dose inhaler, although the bronchodilator capacity was similar. Colforsin dry powder capsules resulted in a measurable bronchodilatation in patients with asthma. PMID- 8422744 TI - Comparative study on the efficacy and acceptability of two contraceptive pills administered by the vaginal route: an international multicenter clinical trial. AB - The efficacy and acceptability of two widely used oral contraceptive tablets, one containing 250 mg levonorgestrel and 50 micrograms ethinyl estradiol and the other containing 150 micrograms desogestrel and 30 micrograms ethinyl estradiol, administered by the vaginal route were compared in 1055 women studied over 12,630 woman-months of vaginal contraceptive pill use. This multicenter clinical trial was performed in nine countries of the developing world by the "South to South Cooperation in Reproductive Health," an organization founded by scientists from the Third World working in the area of reproductive health, and the study was developed and coordinated by one of these centers. The findings of this study confirm the efficacy of both these tablets when administered by the vaginal route. Involuntary pregnancy rates at 1 year of 2.78 for subjects in the levonorgestrel group and 4.54 for subjects the desogestrel group showed no statistically significant difference between the two groups. However, total discontinuation rates of 47.01 for subjects in the levonorgestrel group and 56.33 for subjects in the desogestrel group showed a statistically significant difference between the two groups, and discontinuation rates attributable to prolonged bleeding of 0.6 for subjects in the levonorgestrel group and 3.2 for subjects in the desogestrel group were also significantly higher in the group of subjects using the desogestrel vaginal contraceptive pill. Blood pressure remained at admission values throughout treatment. A statistically significant weight increase from admission values occurred in both groups of subjects. PMID- 8422746 TI - Genetically determined drug-metabolizing activity and desipramine-associated cardiotoxicity: a case report. PMID- 8422748 TI - Annual Meeting of the Western Section American Federation for Clinical Research. Carmel, California, February 17-20, 1992. Abstracts. PMID- 8422747 TI - Biotechnology and therapeutics. PMID- 8422749 TI - Detection of human chromosomes in somatic cell hybrids by PCR analysis. AB - The detection of human chromosomes in somatic cell hybrids is usually made by chromosomal analysis, Southern blot analysis with human probes, and starch-gel electrophoresis of isoenzymes. We describe here a new, quick, and very efficient method to detect human chromosomes in somatic cell hybrids between human and rodent (rat and mouse) cells. The method is based on the polymerase chain reaction to promote amplification of human DNA, using primers derived from localized genes or DNA fragments from each human chromosome. PMID- 8422750 TI - Translocations between two specific human chromosomes detected by three-color "chromosome painting". AB - Translocations between two specific chromosomes are important markers for many human malignancies. Previously, the detection of translocations involving random breakpoints between two specific chromosomes could only be accomplished by banding techniques, which are severely labor intensive and require highly trained technicians. The three-color chromosome painting approach described in this paper was developed in our laboratory to detect translocations between two specific human chromosomes rapidly and accurately, while simultaneously revealing the nonhybridized chromosomes. Because this method efficiently detects translocations involving breakpoints anywhere on the targeted chromosomes, it is ideal as a screening tool for chromosome-specific translocations. PMID- 8422751 TI - A novel, convenient, and inexpensive approach for deriving ISCN (1985) relative lengths: validation by a morphometric study of 100 karyotyped metaphase cells. AB - The present study stemmed from a need for a rapid means of deriving reproducible chromosome measurements. An internal set of standards can serve as the basis for routine, easy, and reliable morphometric comparisons. In this study, a total of 100 karyotyped metaphases were analyzed using the Nestler Run-Mate, a computerized curvilinear measuring tool. The null hypothesis tested was that there are no significant differences between chromosomal relative-length values obtained via this previously untested approach and those cited in ISCN (1985). The results indicate that this new method is not only feasible and adequate but has advantage over the conventional approach, which requires the use of a projector and screen to measure chromosomes in unkaryotyped metaphase spreads; further, it is less expensive and easier than using computerized digitizing tablets, a conclusion supported by time-and-effort measurements. Immediately obvious applications include routine use in clinical cytogenetics laboratories, as well as for fractional length estimations in fluorescent in situ hybridization studies performed in research laboratories that do not have access to expensive automated instrumentation. PMID- 8422752 TI - Localization of the human cystatin D gene (CST5) to chromosome 20p11.21 by in situ hybridization. AB - The gene coding for cystatin D (CST5), a cysteine proteinase inhibitor, was mapped by fluorescent in situ hybridization to human chromosome 20p11.21. This assignment, together with previous data on mapping of members of the cystatin gene family, indicates that cystatin family II genes are all clustered on the short arm of chromosome 20, whereas cystatin family I and III genes are located on the long arm of chromosome 3. PMID- 8422753 TI - Localization of the human oncostatin M gene (OSM) to chromosome 22q12, distal to the Ewing's sarcoma breakpoint. AB - Using fluorescence in situ hybridization, a cosmid clone containing the gene for oncostatin M (OSM) was mapped to human chromosome 22q12, placing the OSM gene in the same chromosome band as the leukemia-inhibitory factor gene (LIF). The location of the OSM gene was determined relative to the t(11;22)(q24;q12) of Ewing's sarcoma and found to be distal to the translocation breakpoint on chromosome 22. Analysis of physical distances by pulsed-field gel electrophoresis demonstrated further that the two genes lie within 500 kb of each other. PMID- 8422754 TI - Localization of the human collagen gene COL7A1 to 3p21.3 by fluorescence in situ hybridization. AB - An 8-kb genomic probe, containing 34 collagen-encoding exons, was localized to 3p21.3 by fluorescence in situ hybridization. The genomic probe encoded a previously uncharacterized carboxyl terminal portion of the alpha 1(VII) collagen chain. This mapping result confirms the previous assignment of the alpha 1(VII) gene (COL7A1) to 3p21 and offers a finer subregional localization than was previously available. PMID- 8422755 TI - Regional assignment of rat androsterone UDP-glucuronosyltransferase gene (UGT2B2) to chromosome 14p21.2-p22. AB - Rat androsterone UDP-glucuronosyltransferase gene (UGT2B2) was regionally assigned to chromosome band 14p21.2-p22 by fluorescence in situ hybridization using a biotinylated cDNA (1.8-kb insert) probe. The present mapping is the first report on the precise localization of the rat UGT2B2 on high resolution banded metaphase chromosomes. PMID- 8422756 TI - Meiotic behavior of rye heterozygous for ditelocentric substitutions. AB - Meiotic behavior has been investigated in rye plants carrying a heterozygous ditelocentric substitution for chromosomes 1R and 3R. Such a chromosomal rearrangement replaces a biarmed chromosome with two telocentrics, a situation karyotypically comparable to a Robertsonian translocation. As in Robertsonian translocation heterozygotes, delayed pairing characterizes the centromeric region in the rye pachytene heteromorphic trivalents. In addition, telocentric axes appear to be frequently engaged in nonhomologous synapsis (e.g., fold-back loops and two-by-two heterosynapsis) at their centromeric ends. Unlike those in Robertsonian translocation heterozygotes, the synaptic abnormalities detected here cannot be due to lack of partner homology. Rather, the chromosomal rearrangement itself appears to interfere with normal synaptonemal complex (SC) progression. The synaptic irregularities, which often result in complex pachytene multivalents, seem to have no effect on chiasma formation, since the rearranged pairs behave properly at metaphase I. Finally, differences in both SC length and progression patterns between plants and animals could explain the trans configurations of telocentric centromeres in plants, whereas only cis configurations of acrocentric kinetochores have been described in animals. PMID- 8422757 TI - Reassessment of breakpoints in chromosome 11p15. AB - Specific tumor-associated rearrangements involving the regions 11p13 and 11p15 have been extensively documented. However, cytogenetic definition of the breakpoints occurring at the boundaries of these two regions was not precise enough to correlate with the molecular data. Using probes corresponding to the genes coding for MYOD1, CTSD, LDHA, and RBTN1 and to the anonymous sequence D11S776, we have reassessed the breakpoints of three hybrids (J1.10, BID7, and NYX3.1) and confirmed the localization or more precisely mapped these four genes and the anonymous DNA marker on different subregions of 11pter-->p13, including the smallest region of 11p15.5 duplicated in a patient with Beckwith-Wiedemann syndrome. PMID- 8422758 TI - Linkage between two blood-group markers in rhesus monkeys (Macaca mulatta). AB - Using marker data on an extensive pedigreed population from the Wisconsin Regional Primate Research Center, we report the first genetic linkage in rhesus monkeys (Macaca mulatta). Five blood-group markers (G, I, J, K, and Q) and one serum protein marker (transferrin) were analyzed. Linkage analyses indicate the existence of loose linkage between G and Q (maximum lod score, Zmax = 4.84 at theta m = 0.314 +/- 0.38, theta f = 0.266 +/- 0.054). The genetic linkage described here constitutes the initial step toward the description of a genetic map in this species. PMID- 8422759 TI - An irradiation-reduced hybrid panel for fine-structure mapping of the Xq28 region in the human genome. AB - Irradiation-reduced somatic cell hybrids containing fragments of the human X chromosome were constructed. Analysis of 16 hybrids that retained the Xq28 region with 12 Xq28-specific markers identified at least six different breakpoints, supporting the order cen-DXS304-DXS374-(DXS33, DXS134, DXS52, DXS15)-RCP-(DXS254, G6PD, F8C)-(DXS115, DXYS64)-qter. The generated panel of hybrids provides a useful tool for fine mapping of probes in the Xq28 region. PMID- 8422760 TI - Demonstration of replication patterns on histone-depleted chromosomes. AB - A modified immunologic technique is described for the purpose of demonstrating replication patterns on mammalian chromosomes after partial histone depletion. Replication patterns were induced by BrdU substitution and visualized by BrdU antibodies, coupled with peroxidase (diaminobenzidine/H2O2) or immunogold detection. The replication patterns obtained by this technique did not reveal any additional details of replication compared to those shown by conventional cytogenetic staining. However, the possibility of demonstrating replication patterns on these partially histone-depleted chromosomes may prove useful for chromosomal in situ hybridization studies since the chromosomes produced are considerably larger than those seen in conventional preparations. PMID- 8422761 TI - Local skin-fold thickness as a clinical predictor of depot size during basal rate infusion of insulin. AB - OBJECTIVE: To determine the influence of local adiposity on insulin depot size during CSII at basal rate. RESEARCH DESIGN AND METHODS: In 27 diabetic patients, a constant infusion of 125I-labeled Actrapid insulin was given, with U-40 insulin at a rate of 1.12 IU/h in 20 patients, and U-100 at a rate of 1 IU/h in 7 patients. After 16 h of infusion, the steady-state depot size was measured by external counting, and the local skin fold was measured with a Harpenden skin fold caliper. RESULTS: U-40 insulin infusion resulted in a steady-state depot size of 5.1 U (2.1-10.9 U), and a corresponding skin-fold thickness of 17.8 mm (5 34 mm). A positive correlation was found between depot size and skin-fold thickness. A similar correlation was observed with U-100 insulin. CONCLUSIONS: During basal rate CSII, large variations in local skin-fold thickness create large variations in the steady-state depot size, which is partly predictable just by lifting the skin fold. PMID- 8422762 TI - Restoration of fuel homeostasis in IDDM patients during pregnancy by an open-loop insulin infusion system. AB - OBJECTIVE: To characterize the effects of subcutaneous insulin pump therapy on the metabolic response (CHO and lipid oxidation, and nonoxidative glucose metabolism) to a glucose challenge of diabetic women at early pregnancy. RESEARCH DESIGN AND METHODS: Seven nondiabetic and seven IDDM pregnant patients on insulin pump therapy were studied at the first trimester. Fuel oxidation rates were determined by indirect calorimetry, and blood levels of substrates and hormones were measured before and for 2 h after ingestion of a 50 g oral glucose load. RESULTS: The increments in npRQ and CHO oxidation rates after the glucose meals in the diabetic women on insulin pump therapy were similar to those in the normal subjects. The glucose disposal data during the 2 h of the studies revealed that the amounts of oxidative and nonoxidative glucose utilization in the control subjects and in the IDDM patients on insulin pump therapy did not show significant differences. CONCLUSIONS: This investigation demonstrated that the treatment of IDDM patients during early pregnancy by an open-loop insulin infusion system is sufficient to normalize their glucose-processing capability with respect to cellular oxidative and nonoxidative glucose metabolism in response to an oral glucose challenge, but some abnormalities in their blood profiles of glucose, lactate, and pyruvate persisted. PMID- 8422763 TI - The effect of glyburide on beta-cell sensitivity to glucose-dependent insulinotropic polypeptide. AB - OBJECTIVE: To assess whether treatment with glyburide alters beta-cell sensitivity to GIP in NIDDM patients. RESEARCH DESIGN AND METHODS: We studied 5 untreated NIDDM patients in a meal study (Ensure, 240 ml/M2) and a 2-h hyperglycemic glucose clamp study (glucose 5.4 mM above fasting). From 60 to 120 min of the clamp, GIP was infused in a primed continuous manner at a rate of 2 pmol.kg-1 x min-1. Subjects then were treated with glyburide. After they had been on a stable dose of medication for 1 mo, the meal study and glucose clamp studies were repeated. RESULTS: In response to treatment, a decrease in fasting glucose and an increase in weight was observed (12.8 +/- 1.8 vs. 8.5 +/- 0.8 mM and 74.3 +/- 6.3 vs. 76.1 +/- 6.3 kg, respectively, P < 0.05). In response to the meal study, the AUC for glucose was less, for insulin was increased, and for GIP was unchanged after treatment (16.9 +/- 2.1 vs. 12.6 +/- 6.9 mM, P < 0.05; 161 +/- 47 vs. 242 +/- 60 pM, P < 0.05; and 199 +/- 22 vs. 219 +/- 18 pM, respectively). During the hyperglycemic clamp, steady-state glucose and 90- to 120-min GIP values were equivalent before and after treatment (18.0 +/- 1.3 vs. 18.3 +/- 1.3 mM and 302 +/- 59 vs. 298 +/- 37 pM, respectively). The 90-120 min insulin responses to the hyperglycemic clamp were greater after therapy (123 +/- 37 vs. 283 +/- 80 pM, P < 0.05) reflecting increased beta-cell responses to GIP. CONCLUSIONS: We conclude that glyburide enhances beta-cell sensitivity to GIP. PMID- 8422765 TI - Decrease of glomerular hyperfiltration in short-term diabetic adolescents without microalbuminuria. AB - OBJECTIVE: This study has been designed to follow prospectively the GFR and UAE of young patients with short-term IDDM and normal UAE. RESEARCH DESIGN AND METHODS: The study population consisted of 19 patients with glomerular hyperfiltration and 19 patients with normal GFR, matched for duration of diabetes and age. GFR has been assessed by radioisotopic tracer and UAE by RIA at the beginning of the study and after 30.5 +/- 10.4 mo of follow-up. RESULTS: GFR decreased in the two groups btt delta GFR of patients with glomerular hyperfiltration was greater than delta GFR of patients with normal GFR (0.83 +/- 0.55 vs. 0.28 +/- 0.63 ml.min-1.mo-1; P < 0.01). UAE, BP, and prevalence of microalbuminuria were comparable between the two groups at follow-up. Rate of fall of GFR was positively correlated with initial GFR (r = 0.59, P < 0.001) but not with initial UAE, BP, or changes in HbA1C, UAE, BP, or pubertal development during follow-up. CONCLUSIONS: Investigation of kidney function in children and adolescents with IDDM over a 3-yr follow-up period shows that glomerular hyperfiltration is characterized by a greater decline in GFR without an increased rate of appearance of microalbuminuria, than in patients with normal GFR. PMID- 8422764 TI - Glomerular hyperfiltration in NIDDM patients without overt proteinuria. AB - OBJECTIVE--To evaluate the frequency and correlates of glomerular hyperfiltration in NIDDM patients without overt proteinuria. RESEARCH DESIGN AND METHODS--A cross sectional study was conducted. Seventy-one consecutive NIDDM patients attending an outpatient clinic, with Albustix-tested negative urine and a 24-h AER < 200 micrograms/min, were examined for long-term complications of diabetes. We measured their GFR (51Cr-EDTA single-injection method), 24-h AER (RIA), plasma creatinine, HbA1c, total cholesterol, triglycerides, urinary glucose, and urea. RESULTS--GFR above the upper limit of the normal range for age-matched control subjects (137.1 ml.min-1 x 1.73 m2) was present in 15 of 71 (21%) NIDDM patients. Subjects with normal and hyperfiltration did not differ in terms of age, sex distribution, BMI, duration of NIDDM, BP, AER, or frequency of long-term complications. Plasma glucose was significantly higher in subjects with hyperfiltration (mean [range]: 12.8 [4.3-18.7] vs. 8.7 [2.6-17.5] mM). HbA1c failed to reach statistical significance, although it tended to be higher in the group with hyperfiltration (10.4 [6.7-13.9] vs. 9.4 [4.2-16.5]%, P = 0.10). Age (rS -0.37, P = 0.002), FPG (rS 0.45, P < 0.0005), total cholesterol (rS -0.31, P = 0.008), and glycosuria (rS 0.40, P = 0.001) correlated significantly with GFR. In a stepwise multiple regression analysis, FPG, age, and total cholesterol emerged as significant correlates of the dependent variable GFR. CONCLUSIONS- Hyperfiltration occurred in 21% of NIDDM patients without overt proteinuria. FPG and age significant correlates of the GFR in these patients. Cholesterol is significantly (although only modestly) correlated with the GFR. PMID- 8422766 TI - Determinants of clinical remission in recent-onset IDDM. AB - OBJECTIVE: To assess the relationship of SI and insulin secretion (C-peptide levels) to remission status in recent-onset IDDM. RESEARCH DESIGN AND METHODS: We followed 22 newly diagnosed patients, of whom 16 received immunomodulatory treatment with low-dose (5 mg.kg-1 x day-1) CsA and/or short-term (72 h) methylprednisolone and 6 received standard insulin treatment, at 3-mo intervals for 12 mo. Insulin secretion was assessed by C-peptide levels and AIRglu, which was determined as the area under the insulin response curve, above the fasting level, from 0-10 min after a 0.3 g.kg-1 x i.v. glucose bolus. SI was assessed by the minimal model technique applied to a frequently sampled IVGTT. Clinical remission was defined in those patients who maintained normal range GHb and capillary blood glucose levels < 7.8 mM premeal without insulin therapy for a minimum of 14 days. RESULTS: The rate of clinical remission was not different with immunomodulatory treatment; nor were the metabolic parameters of plasma C peptide levels, AIRglu, and SI different in the treatment groups. The mean plasma C-peptide level improved significantly at 3 mo and was maintained to 12 mo. AIRglu was grossly subnormal throughout, but a significant improvement was seen at 3 and 6 mo. Mean SI was normalized at 3 and 6 mo but not maintained beyond 9 mo. The maximum rate of clinical remission was seen at 6 mo. CONCLUSIONS: Clinical remission in recent-onset IDDM patients is associated with improvement in both insulin secretion and SI. Although the improvement in basal C-peptide persisted, AIRglu increased only transiently and declined as loss of remission occurred in most patients. Loss of remission to an insulin-requiring state is associated with a decrease in SI. PMID- 8422767 TI - Sex differences in incidence of IDDM in age-group 15-29 yr. Higher risk in males in Province of Turin, Italy. AB - OBJECTIVE: To report the incidence of IDDM in the age-group 0-29 yr in the Province of Turin, Italy (951, 445 inhabitants 0-29 yr of age), over a 5-yr period (1984-1988) according to age, sex, and geographical region within the area and to identify any temporal trend. RESEARCH DESIGN AND METHODS: The survey used as the primary data source the list of all patients attending diabetic clinics, and as secondary data source, used the list of hospital discharges for diabetes. RESULTS: We identified 298 incident cases of IDDM in people 0-29 yr of age. Estimated completeness of ascertainment of the registry was 97%. Age-adjusted (world-standard) incidence rates were 7.40/100,000 (95% CI 6.28-8.71), 5.83 (4.95 6.86), and 6.70 (5.97-7.51), respectively, in the age-groups 0-14, 15-29, and 0 29 yr. Incidence was significantly higher in males than in females in the age group 15-29 yr (7.36, 6.02-8.98, vs. 4.21, 3.12-5.56). An increasing incidence from rural areas to the greater Turin area (city and its industrial belt) was evident. No significant temporal trend during the study period was found, although year-to-year variability was evident, with the highest incidence in 1984. CONCLUSIONS: This study suggests a high male-to-female ratio of incidence of IDDM after 14 yr; either sex hormones or different exposure to environmental determinants could be involved. PMID- 8422768 TI - Development of macrovascular diseases in NIDDM patients in northern Taiwan. A 4 yr follow-up study. AB - OBJECTIVE: To assess the development of macrovascular diseases and explore major associative factors in NIDDM. RESEARCH DESIGN AND METHODS: A total of 479 NIDDM patients > or = 40 yr of age were recruited from four community primary care health centers of northern Taiwan in July 1986 for a cohort study with a 4-yr follow-up. No patient required insulin therapy within 1 yr of diagnosis nor had a history of diabetic ketoacidosis. All were able to participate independently in the activities of daily living. BP and ECG were measured, and a structured questionnaire was asked of each patient. Venous blood after overnight fasting was collected every year to measure cholesterol, HDL cholesterol, plasma glucose, and HbA1c. RESULTS: The duration of diabetes was associated with the development of stroke with a relative risk of 1.063 for every 1-yr increment (P = 0.07). As for HVDs, the significant risk factors were serum cholesterol and HbA1c. For every 1 mg/dl increase in mean total cholesterol level, the relative risk of developing HVD increased 1.016-fold (P = 0.04). For every 1% increase in HbA1c, the relative risk of developing HVD increased 1.170-fold (P = 0.01). With regard to leg VDs, sex and cigarette smoking were significant risk factors. Women diabetic subjects had a higher relative risk than men. Cigarette smoking was significantly associated with leg VD with a relative risk of 6.9 for smokers compared with nonsmokers. The most significant risk factor for LVD was the total cholesterol level. For every 1-mg/dl increase in mean serum cholesterol level, the relative risk of LVD increased 1.013-fold. CONCLUSIONS: In the prevention of macrovascular diseases, effective intervention of the nondiabetic cardiovascular risk factors may be as important as or even more important than the good control of diabetes. PMID- 8422769 TI - Coronary heart disease risk factors in morbidly obese women with normal glucose tolerance. AB - OBJECTIVE--To examine if the risk for CHD increases progressively with increases in the BMI of normoglycemic, hyperinsulinemic, morbidly obese women (BMI > or = 35 kg/m2). RESEARCH DESIGN AND METHODS--Insulin sensitivity was evaluated by calculating an ISI following an OGTT. There was a significant linear relationship between ISI and BMI fitted by two straight lines intersecting at a point corresponding to a BMI of 29.7 +/- 1.5 kg/m2. Significant linear relationships between insulin sensitivity and BMI were obtained below and above this breakpoint. Similarly, a breakpoint for the relation between dBP and BMI corresponding to a BMI > or = 33.7 +/- 3.4 kg/m2 was obtained. Significant linear relationships between BMI and plasma fasting glucose, triglyceride, cholesterol, HDL cholesterol, sBP, or dBP were not observed in the women with a BMI > 35 kg/m2. RESULTS--Compared with lean (BMI < 27) women of similar age, the morbidly obese patients appear to be at a higher risk for CHD. This is suggested by statistically significant increases in fasting insulin (mean +/- SD; 187 +/- 137 vs. 64.2 +/- 16.2 pM) and triglyceride levels (128 +/- 78.1 vs. 73 +/- 25 mg/dl), sBP (132 +/- 114 vs. 104 +/- 15.8) and dBP (84 +/- 72 vs. 67 +/- 2.1 mmHg), and decreases in HDL cholesterol (1.03 +/- 0.44 vs. 1.29 +/- 0.82 mM) and apo A-I (91 +/- 55 vs. 122 +/- 35 mg/dl) concentrations. CONCLUSIONS--It appears that there may be a threshold of body mass up to which insulin sensitivity is associated with CHD risk. Above this threshold, there does not appear to be a progressive increase in the risk factors for CHD with increases in BMI. PMID- 8422770 TI - Impaired glucose tolerance and its relationship to ECG-indicated coronary heart disease and risk factors among Chinese. Da Qing IGT and diabetes study. AB - OBJECTIVE: To determine the prevalence of IGT and diabetes and identify the occurrence of CVD and its risk factors. RESEARCH DESIGN AND METHODS: This study was a population-based, cross-sectional study of 110,660 residents, 25-74 yr of age of DaQing, Hei Long Jiang Province, China. RESULTS: Using 1985 WHO criteria, 577 (5.5/1000) subjects with IGT and 630 (6.0/1000) with newly diagnosed diabetes were identified. In addition, 190 (1.7/1000) with previously known diabetes were identified. Hypertension, obesity, and abnormal albumin excretion were twice as frequent in those with IGT as in 279 men and 240 women of similar age and sex distribution who had normal OGTTs. Plasma cholesterol and TGs were higher, and HDL cholesterol was lower in subjects with IGT than in nondiabetic subjects. Those with IGT had a prevalence of electrocardiographically recognized CHD 9.5 fold greater than the normoglycemic subjects. Subjects with IGT had higher plasma insulin concentrations, but the 1-h insulin-glucose ratio after the 75-g glucose load was lower. In a forward stepwise multiple logistic regression analysis, IGT itself remained an independent factor associated with CHD after adjustment for age, sex, cigarette smoking, plasma cholesterol, BP, and obesity. CONCLUSIONS: Therefore, in China, IGT may occur with almost as high a frequency as diabetes and is accompanied by an increased frequency of CVD and its risk factors. Estimates from this study indicate that > 12.0% of all ECG-indicated CHD in the Da Qing population occurs in individuals with IGT and NIDDM. The IGT subjects identified in this survey form the cohort for a long-term follow-up and intervention study. PMID- 8422771 TI - Lower-extremity amputations in diabetic and nondiabetic patients. A population based study in eastern Finland. AB - OBJECTIVE: To study the incidence of LEAs attributable to PVD in diabetic and nondiabetic patients. The age at first amputation, the level of amputation, the number of reamputations, and survival after amputation also were examined in the study populations. RESEARCH DESIGN AND METHODS: This retrospective study was based on a population of 253,000 inhabitants in eastern Finland. All patients with their first LEA performed during the period from 1 January 1978 to 31 December 1984 were identified from the registers of operation theaters in the study area. Furthermore, patient records and death certificates were reviewed. Amputations attributable to causes other than evident atherosclerotic vascular disease were excluded. RESULTS: Altogether, 477 patients (85 diabetic men, 127 nondiabetic men, 169 diabetic women, and 96 nondiabetic women) were identified. The overall LEA rate was 26.9/100,000 per yr, and the incidence increased strongly with age in both diabetic and nondiabetic patients. The age-adjusted amputation incidence per yr was 349.1/100,000 for diabetic men, 33.9/100,000 for nondiabetic men, 239.4/100,000 for diabetic women, and 17.2/100,000 for nondiabetic women. The proportion of peripheral (toe, leg) amputations was markedly higher in diabetic patients who also tended to have more reamputations during the follow-up than did nondiabetic subjects. The diabetic status per se was a statistically significant risk factor for mortality in women, but not in men. CONCLUSIONS: Diabetic men and women had a 10.3- and 13.8-fold higher risk, respectively, for LEA. PMID- 8422772 TI - Deranged platelet calcium homeostasis in poorly controlled IDDM patients. AB - OBJECTIVE: Platelet hyperfunction frequently occurs in IDDM. As in many other cellular systems, cytosolic free Ca plays a key role in platelet activation. RESEARCH DESIGN AND METHODS: We measured cytosolic free Ca concentration ([Ca2+]i) by means of the fluorescent probe fura-2 in 60 IDDM patients (mean age 30.8 yr, range 18-50 yr) and in 31 age-matched healthy control subjects. Platelets were studied in both resting conditions and after stimulation with thrombin at 0.05, 0.1, and 0.5 U/ml. RESULTS: No differences were noted between control subjects and diabetic patients, as a whole. Patients with a poor metabolic control (HbA1c > 8%) had significantly (P < 0.01 and P < 0.03) higher [Ca2+]i in resting platelets. The presence or absence of retinopathy did not modify resting platelet [Ca2+]i. After stimulation with thrombin, a significantly (P < 0.009) higher rise of platelet [Ca2+]i was observed only in those patients who were both free from complications and had good metabolic control. A highly significant (P < 0.001) correlation was found between resting [Ca2+]i and both blood cholesterol and HbA1c in the diabetic patients. Platelets from 10 young healthy subjects also were studied after in vitro incubation with various glucose concentrations (from 1.68 to 56 mM): resting and thrombin-stimulated platelet [Ca2+]i and thrombin-induced aggregation were not modified. CONCLUSIONS: These data confirm that platelet hyperfunction is present in IDDM patients who have unsatisfactory metabolic control, and give evidence that such an activation involves Ca homeostasis. Acute variations of blood glucose concentration are probably not influent, in this respect. PMID- 8422773 TI - Successful treatment of severe refractory sulfonylurea-induced hypoglycemia with octreotide. AB - OBJECTIVE: To test the clinical use of octreotide in the treatment of sulfonylurea-induced hypoglycemia. RESEARCH DESIGN AND METHODS: A case is reported of sulfonylurea-induced hypoglycemic coma in a nondiabetic subject, which was complicated by relapse of hypoglycemia after resuscitation with intravenous dextrose. Subcutaneous octreotide, 50 micrograms 12 hourly, suppressed stimulated endogenous insulin secretion, thereby preventing a further recurrence of hypoglycemia. RESULTS: No adverse effects of treatment were observed. CONCLUSIONS: These results suggest a significant role for octreotide as an adjunct to intravenous dextrose in the management of severe and refractory cases of sulfonylurea-induced hypoglycemia. PMID- 8422774 TI - A search for malnutrition-related diabetes mellitus among Ethiopian patients. AB - OBJECTIVE: To search for evidence of MRDM among Ethiopian patients. RESEARCH DESIGN AND METHODS: We reviewed the records from March 1976 to January 1991 of 1835 Ethiopian diabetic patients registered consecutively in the Diabetic Clinic of Yekatit 12 Hospital in Addis Ababa, Ethiopia. RESULTS: Of those aged 15-30 at onset, 41.3% had a BMI < 19 kg/m2 at diagnosis, and 55% of the latter had a history of normal weight before the onset of symptoms. The frequency of KA was similar in the very thin patients and others of the same age-group, and only 1 patient required > 1.5 IU insulin in 24 h. Pancreatic calcification was observed in only 4 middle-aged men with histories of alcohol abuse, and the very thin patients did not have other stigmata of malnutrition. Of the 1604 patients > or = 18 yr of age at diagnosis, only 21 of 1116 who knew their previous weight had a BMI < 19 kg/m2 before the onset of diabetic symptoms, and most of the 178 patients with a BMI < 19 kg/m2 on treatment had been normal or even overweight. In regression analysis, the factors associated with weight loss before diagnosis were the duration of symptoms in type I diabetes patients and the need for insulin from diagnosis, poverty certification, and symptom duration in type II diabetes patients. Weight gain with treatment was related to female sex, the duration of symptoms, and the absence of tuberculosis in type I diabetes patients and related to an address/birthplace not in Addis Ababa in type II diabetes patients. CONCLUSIONS: The undernutrition at presentation is probably caused by the untreated diabetic state and is reversible with treatment, even if the patient is poor and/or lives in a rural area. No convincing cases of MRDM fulfilling the published definition could be found. PMID- 8422775 TI - Behavioral treatment of obesity. Its application to type II diabetes. AB - This review describes the improvements made in the behavioral treatment of obesity from the 1970s to the present, and then provides a comparable overview of progress that has been made in the behavioral treatment of obese patients with type II diabetes by Wing et al. (13-19) at the University of Pittsburgh. Evidence is presented to show that structured exercise, VLCDs, and intensification of treatment programs may be useful in improving the long-term outcome of behavioral weight-loss interventions. PMID- 8422776 TI - Strategies for improving maintenance of weight loss. Toward a continuous care model of obesity management. AB - The management of obesity represents an important objective in the care of many NIDDM patients. In recent years, progress has been made in increasing initial weight reductions, but poor long-term maintenance of weight loss remains a vital clinical concern. This article reviews the challenge of weight-loss maintenance and recommends the adoption of a continuous care model of obesity management. Strategies to improve the long-term maintenance of weight loss are described, and empirical tests of their effectiveness are reviewed. Collectively, the findings suggest that, after treatment for obesity, multifaceted programs comprised of continued professional contact, skills training, social support, and exercise, can enhance the long-term maintenance of weight loss. PMID- 8422777 TI - Intensive conventional insulin therapy for type II diabetes. Metabolic effects during a 6-mo outpatient trial. AB - OBJECTIVE: To determine whether tight glycemic control can be obtained using intensive conventional split-dose insulin therapy in the outpatient management of type II diabetes without development of unacceptable side effects. RESEARCH DESIGN AND METHODS: Fourteen type II diabetic subjects were treated with an intensive program of conventional insulin (subcutaneous NPH and regular insulin before breakfast and supper) for 6 mo. Insulin dose adjustments were based on an algorithm built on frequent CPG measurements (4-6 times/day). Patients were monitored biweekly as outpatients and admitted 1 day/mo for metabolic evaluation. RESULTS: Glycemic control was achieved by 1 mo (mean plasma glucose fell from 17.5 +/- 0.9 to 7.7 +/- 0.7 mM, P < 0.001) and remained in this range thereafter. Hypoglycemic events at 1 mo were infrequent (mean +/- SE events per patient per month: 4.1 +/- 0.3) and mild in nature, and progressively decreased to 1.3 +/- 0.5 events/mo by 6 mo. After treatment, basal HGO fell 44% from 628 +/- 44 to 350 +/- 17 mumol.m-2.min-1 (P < 0.001), and maximal rates of glucose disposal measured by hyperinsulinemic euglycemic clamp (1800 pmol.m-2.min-1) improved from 1418 +2- 156 to 1657 +/- 128 mumol.m-2.min-1 (P < 0.05). The total dose of exogenous insulin required was 86 +/- 13 U at 1 mo and 100 +/- 24 U at 6 mo. During treatment, mean serum insulin levels increased from 308 +/- 80 to 510 +/- 102 pM (P < 0.05), while body weight increased from 93.5 +/- 5.8 to 102.2 +/- 6.8 kg (P < 0.001). Both pre- and posttreatment glucose disposal rates correlated with the total exogenous insulin dose required to achieve glycemic control (r = 0.75 and -0.78, both P < 0.005). Weight gain was inversely related to the pretreatment glucose disposal rate (r = -0.53, P < 0.05) and directly correlated with both mean day-long serum insulin level (r = 0.67, P < 0.01) and total exogenous insulin dose (r = 0.62, P < 0.02). CONCLUSIONS: Intensive CIT, when combined with CBG measurements, can be used to rapidly improve glycemic control in type II diabetes without development of unacceptable hypoglycemia. This degree of metabolic improvement, however, requires large doses of exogenous insulin to overcome peripheral insulin resistance and results in greater hyperinsulinemia with progressive weight gain. PMID- 8422778 TI - Diabetes in American Indians and Alaska Natives. PMID- 8422779 TI - Determinants of diabetes mellitus in the Pima Indians. AB - OBJECTIVE: To review the research findings on the determinants of diabetes mellitus in Pima Indians. RESEARCH DESIGN AND METHODS: Pima Indians in Arizona have participated in a longitudinal diabetes study that has provided data on and hypotheses about the development of NIDDM. Findings from this study are reviewed and updated. RESULTS: Frequency distributions of plasma glucose and HbA1 are bimodal in Pima adults, and substantial risk of the specific vascular complications of diabetes is confined to those in the higher components of these distributions. These findings contributed to the adoption of internationally recognized criteria for classification of glucose tolerance. Diabetes in the Pimas is strongly familial, and probably of genetic origin, although the precise nature of the gene or genes involved remains unknown. Obesity, which is at least in part environmentally determined, is a major factor interacting with the presumed genetic susceptibility to result in diabetes. The incidence of diabetes in the Pimas has increased during the last several decades, providing further evidence for environmental-genetic interaction. Longitudinal studies suggest that the progression from normal to diabetes can be considered to involve two stages. The first, primarily attributable to insulin resistance, leads to impaired glucose tolerance, and the second, which depends on insulin secretory failure, leads to worsening hyperglycemia and overt diabetes. CONCLUSIONS: The Pimas and many other American Indian populations suffer from a high incidence of diabetes and its characteristic disabling or fatal complications, and would benefit from continued research on the pathogenesis and prevention of the disease. PMID- 8422780 TI - Insulin resistance in the pathogenesis of NIDDM in Pima Indians. AB - NIDDM in Pima Indians is characterized by obesity, abnormal insulin secretion, insulin resistance, and excess hepatic glucose output. Cross-sectional studies, and, as yet incomplete longitudinal studies of nondiabetic and diabetic Pima Indians suggest that the natural history of the disease begins with insulin resistance and, subsequently, when insulin secretion fails, increasing hepatic glucose output occurs, resulting in increasing fasting hyperglycemia. The insulin resistance that precedes the development of fasting hyperglycemia is not due solely to obesity. Insulin resistance aggregates in families and the trimodel frequency distribution of insulin action in vivo suggests it may have genetic determinants. PMID- 8422781 TI - Energy metabolism in obesity. Studies in the Pima Indians. AB - OBJECTIVE: To review the major data collected over the past 8 yr regarding energy expenditure in relationship to obesity and the development of obesity in the Pima Indian population. RESEARCH DESIGN AND METHODS: The different components of 24-h energy expenditure (i.e., the RMR, the thermic effect of food, and the energy cost of spontaneous physical activity) were measured in a respiratory chamber after a few days on a weight-maintenance diet. RESULTS: Fat-free body mass, the major determinant of RMR, explains 82% of its variance. However, an extra 7% of the variance in RMR observed between people is related to family membership. The variability of RMR for a given body size and composition is of importance, because a low metabolic rate is a major risk factor for weight gain in man. Also, genetic factors seem to be the cause of the familial aggregation of metabolic rate in man. CONCLUSIONS: The high prevalence of obesity and NIDDM in the Pima Indian community might be the consequence of a "thrifty genotype." The increasing evidence that obesity cannot always be attributed to gluttony and sloth forces us to consider obesity as a "real metabolic disease" that needs to be treated as such, using new behavioral and pharmacological therapies. PMID- 8422782 TI - Diabetes in American Indians. An overview. AB - OBJECTIVE: To review the growth of diabetes as a major health problem for American Indians and Alaska Natives. RESEARCH DESIGN AND METHODS: Pertinent studies of diabetes in Indians previously published are reviewed and compared with current data. RESULTS: Although diabetes rates may vary among different Indian tribes, diabetes has become a widespread health problem. CONCLUSIONS: Our understanding of the natural history of diabetes among Indians has improved, but better strategies to prevent complications and ultimately to prevent diabetes are urgently needed in Indian communities. PMID- 8422783 TI - Diabetes and its complications among selected tribes in North Dakota, South Dakota, and Nebraska. AB - OBJECTIVE: To determine the prevalence and incidence rates of diabetes and two specific complications for selected American-Indian tribes in North Dakota, South Dakota, and Nebraska. RESEARCH DESIGN AND METHODS: A descriptive epidemiological study was conducted using ambulatory care data during 1987 for prevalence and diabetes registries and complication case reporting during 1988 from IHS facilities on reservations in these states. RESULTS: The Winnebago and Omaha tribes had the highest age-adjusted diabetes rates, with prevalence 8.8 times and incidence 7.7 times the respective U.S. rates. The diabetes prevalence rate of combined data for the Sioux was 3.7 times the U.S. rate. Among Sioux Indians, the age-adjusted incidence rate for ESRD was 4.8 times the American-Indian/Alaska Native rate and 13.4 times the rate for U.S. whites. The proportion of new diabetes-related ESRD (86%) was almost 3 times greater than the general U.S. population rate (30%). Also, among the Sioux, the age-adjusted incidence rate for LEA (86.7/10,000 diabetic population) was 1.5 times higher than the U.S. rate; the proportion of diabetes-related LEA (84%) was 1.8 times higher than the general U.S. population rate (45%). CONCLUSIONS: The age-adjusted rates of diabetes and certain complications among these Northern Plains tribes are greater than the U.S. rates. Improved health services to detect and monitor diabetes and its complications and community-based prevention activities directed at the epidemic of diabetes among the various Indian tribes are urgently needed. PMID- 8422784 TI - Increasing prevalence of diabetes among the three affiliated tribes. AB - The findings from a 1988 audit of medical records of the Three Affiliated Tribes (the Mandan, Arickara, and Hidatsa) at the Fort Berthold, ND, Indian Reservation were compared with the findings from a 1975 audit. The total population served by the IHS clinic there has remained about the same. However, the number of people diagnosed as having diabetes has increased by > 40%. This increase in known cases is caused largely by an increase in the number of women aged > 40 yr who have diabetes. The actual prevalence of diabetes among people aged > 40 yr has increased very little, presumably because the clinic population has aged somewhat over this period of time. PMID- 8422785 TI - Diabetes in Mississippi Choctaw Indians. AB - OBJECTIVE: To establish a registry of patients with diabetes and determine the point prevalence of diabetes in the Mississippi Choctaw Indians at the end of September 1989. RESEARCH DESIGN AND METHODS: A survey of diabetes cases was conducted, using the NDDG criteria for diagnosis with comparison to U.S. rates determined from the 1979-1981 NHIS. IHS-supported hospital and outpatient clinics serving the Choctaw Indians of Mississippi were the setting. Medical charts of patients on the computer record of outpatient visits seen for diabetes and laboratory lists of patients who had elevated blood glucose levels in 1988 were reviewed. RESULTS: The age-adjusted prevalence of 163.2/1000 population was 6.5 times (95% CI 4.3-9.8) the U.S. rate. The prevalence of diabetes in the Choctaw Indians was significantly higher than the U.S. rates for all age-groups except for those < 15 yr of age. CONCLUSIONS: Intervention programs are urgently needed to prevent or delay the onset of diabetes and its complications in this southeastern tribe. The registry system is being used to gather information on the prevalence and incidence of diabetic complications and associated risk factors to guide programs. PMID- 8422786 TI - Prevalence of diabetes and its complications in the Eastern Band of Cherokee Indians. AB - OBJECTIVE: To determine the prevalence of diabetes and selected complications among the Eastern Band of Cherokee Indians in North Carolina. RESEARCH DESIGN AND METHODS: Multiple IHS data systems were used to determine diabetes prevalence and complication rates. The RPMS was used to identify diabetes cases as of April 1989, degree of Indian inheritance, cardiovascular diseases (including hypertension), and retinopathy. Data on laser treatments and LEA were obtained from individual registries. Information on ESRD was obtained by a research assistant. The IHS Ambulatory Patient Care reporting system was used to calculate the number of diabetic patients for the years 1982-1987. The IHS user population was used as the denominator. Rates were age-adjusted to the 1980 U.S. population and diabetic population by the direct method. RESULTS: Using clinical records, the age-adjusted prevalence of diabetes in 1988 was 105.6/1000 people, four times the U.S. rate. Rates of diabetes were highest in the groups with the highest degree of Indian inheritance. LEAs occurred among diabetic patients at three times the rate for the U.S. Between 1985 and 1989, new cases of ESRD occurred at a crude annualized rate of 578/million, approximately six times the rate for U.S. whites. CONCLUSIONS: Diabetes presents a major burden to the Eastern Band of Cherokees. The limitation inherent in this study probably result in underestimation of the prevalence of diabetes and its complications in the community. Future studies are needed to assess the impact of medical and preventive programs on diabetes prevalence and complications in this community. PMID- 8422787 TI - Diabetes prevalence, incidence, and complications among Alaska Natives, 1987. AB - OBJECTIVE: To provide descriptive epidemiological data on diabetes mellitus among Alaska Natives, including incidence, updated prevalence, and incidence rates of ESRD, LEA, MI, and stroke in the diabetic population. RESEARCH DESIGN AND METHODS: In IHS and tribal contract hospitals and clinics throughout Alaska, Alaskan Indians, Eskimos, and Aleuts living in Alaska with documented diabetes mellitus were studied from 1986 to 1987. RESULTS: The number of diabetic patients increased from 610 to 708, and the prevalence changed from 15.7 to 17.4/1000 (not a statistically significant increase). Incidence rates per 10,000 diabetic person yr for complications were 38 for ESRD, 69 for LEA, 92 for MI, and 92 for stroke. CONCLUSIONS: The prevalence of diabetes mellitus increased during the period of observation, but not to a statistically significant degree. Incidence rates for diabetes are lower than for the U.S. general population, but complications rates are as high as those in other diabetic populations. PMID- 8422788 TI - Diabetes in St. Regis Mohawk Indians. AB - OBJECTIVE: To establish a registry of diabetes patients and to determine the point prevalence of diabetes in the St. Regis Mohawk New York community at the end of August 1989. RESEARCH DESIGN AND METHODS: Participants in this study consisted of residents of the St. Regis Mohawk reservation in New York state, who were registered for health care at the St. Regis Mohawk Clinic. This study was a survey of diabetes cases using the National Diabetes Data Group criteria for diagnosis with comparison with the U.S. rates determined from the 1987-1982 National Health Interview Survey and Canadian Mohawk Indian rates from other studies. RESULTS: The age-adjusted prevalence of 48.8/1000 Mohawk population was two times (95% CI 1.2-3.2) the U.S. rate. Although precise comparisons with Canadian studies are difficult because of differences in study design, rates for Quebec Mohawks aged 45-64 yr were similar. CONCLUSIONS: A high prevalence (21%) among New York Mohawk Indians < 45 yr of age, compared with 17% of diabetic patients in that age-group in the U.S. population, means that complications resulting from diabetes will occur at a relatively young age, placing an increased burden on the health-care system with costs of patient care associated with diabetes increasing accordingly. The registry system is being used to gather information on the prevalence and incidence of diabetic complications and risk factors for developing diabetes. These data will be used to evaluate programs. PMID- 8422789 TI - Prevalence of diagnosed diabetes and selected related conditions of six reservations in Montana and Wyoming. AB - OBJECTIVE: To describe the prevalence of diabetes and related complications in Plains Indians. RESEARCH DESIGN AND METHODS: The medical records of individuals with diagnosed diabetes seen in IHS facilities before 1 January 1987 were reviewed. Records were obtained from primary care clinics and hospitals accessible to the following six reservation communities in Montana and Wyoming: Blackfeet, Crow, Fort Belknap, Fort Peck, Northern Cheyenne, and Wind River. Medical records were reviewed to ascertain accuracy of diagnosis and associated complications. RESULTS: Age-adjusted prevalence rate of diagnosed diabetes was 119/1000 adults > or = 15 yr of age. CONCLUSIONS: Diabetes rates in Plains Indians are higher than rates described for the general U.S. population. PMID- 8422790 TI - Diabetes in a northern Minnesota Chippewa Tribe. Prevalence and incidence of diabetes and incidence of major complications, 1986-1988. AB - OBJECTIVE: To determine the prevalence and incidence of diabetes, and the incidence of major diabetic complications, in a Chippewa Indian population. RESEARCH DESIGN AND METHODS: The format was a longitudinal population study that used active community and health center-based surveillance. The setting was a North American Indian reservation community of 4075 residents, served by an IHS clinic from 1986 to 1988. Patients were American Indians of Chippewa descent living on or near the Red Lake Reservation. RESULTS: Midway through the study, 346 people had been diagnosed with diabetes, which yielded an age- and sex adjusted point prevalence of 148/1000 population. The adjusted rate for individuals > or = 25 yr of age was 252/1000 population, 3.82 times the U.S. rate (CI 2.95-4.93). Some 97 new cases of diabetes were identified for an age- and sex adjusted average annual incidence of 17/1000 population. The incidence of hospitalization for LEA was 26/1000 diabetic person-yr, 4.3 times the 1978 U.S. rate (95% CI 2.8-6.8). Twelve individuals developed proliferative retinopathy, for an incidence of 12/1000 diabetic person-yr. Newly diagnosed ESRD incidence was 6/1000 diabetic person-yr. Twenty-three acute myocardial infarctions were observed, yielding an incidence of 22/1000 diabetic person-yr. CONCLUSIONS: Diabetes and its complications are prevalent in this Chippewa population, and further surveillance is needed to evaluate the effectiveness of prevention efforts. PMID- 8422791 TI - Prevalence of diagnosed diabetes among American Indians and Alaska Natives, 1987. Estimates from a national outpatient data base. AB - OBJECTIVE: To estimate the prevalence of diagnosed diabetes among American Indians and Alaska Natives served by the IHS. RESEARCH DESIGN AND METHODS: This was a cross-sectional study of the 1987 IHS national outpatient data base. RESULTS: Prevalence rates of diagnosed diabetes determined from the IHS outpatient data base were consistent with recent studies of diabetes in different IHS areas. IHS-wide, age-adjusted prevalence was 69/100,000, or 2.8 times the U.S. rate. There was considerable variation in prevalence rates of diabetes throughout the country, with rates ranging from 15.3/100,000 in Alaska to 119.2/100,000 in southern Arizona. CONCLUSIONS: This study documented the high prevalence of diabetes among American Indians and Alaska Natives and the wide variation in rates between different tribal groups. This study also demonstrated the feasibility of using an outpatient data base to estimate rates of disease that have uniform methods of diagnosis and result in frequent clinic visits. PMID- 8422792 TI - Cardiovascular disease among American Indians and Alaska Natives. AB - OBJECTIVE: To compare the regional differences in cardiovascular disease in AI/AN with the U.S. general population and determine the parity gap and preventable proportion of cardiovascular mortality. RESEARCH DESIGN AND METHODS: Age-adjusted cardiovascular disease mortality rates for 1981-1983 and hospital discharge rates for 1982-1984 reported by the IHS were compared with U.S. data for 1982 and 1983, respectively. RESULTS: Rates of ischemic heart disease and atherosclerosis were found to be generally low among AI/AN although those in the 25- to 44-yr age group have higher death rates from cardiovascular disease than in the U.S. population. Although the mortality rate from cardiovascular disease in AI/AN is 19% lower than the rate for the general U.S. population, the parity gap in individual regions of the U.S. ranges from favorable to extremely unfavorable. There were also wide variations in the preventable gap theoretically possible by reduction of the three major risk factors. CONCLUSIONS: Changing nutrition and exercise patterns and the increasing prevalence of diabetes in many Indian tribes may have adverse effects in the future, possibly increasing the prevalence of heart disease. Regional differences in the prevalence of some major cardiovascular risk factors (smoking, hypertension, hypercholesterolemia, and diabetes) are the probable explanation for these differences in cardiovascular morbidity and mortality rates. Prevention and treatment of these risk factors will have the greatest impact in attempts to reduce cardiovascular disease among AI/AN. In addition, moderation in the use of alcohol, or abstinence, may prevent sudden deaths resulting from acute intoxication. PMID- 8422793 TI - Diabetes and plasma lipoproteins in Native Americans. Studies of the Pima Indians. AB - OBJECTIVE--To determine the effects of diabetes on plasma lipoproteins in Pima Indians, to identify metabolic determinants of these differences, and to examine the effects of various modes of diabetes therapy. RESEARCH DESIGN AND METHODS--A series of studies was performed including a population survey of lipoprotein concentrations, kinetic studies of VLDL and LDL metabolism, and studies of the effects of weight loss, sulfonylureas, and high carbohydrate diets. RESULTS- Population data established that diabetes was consistently associated with elevations in total and VLDL TGs and decreases in HDL cholesterol. Metabolic studies indicated multiple alterations in VLDL metabolism induced by NIDDM, including overproduction of VLDL TG, impaired clearance of VLDL TG and apoB, and decreases in adipose tissue lipoprotein lipase. Although changes in LDL concentrations accompanying NIDDM were minimal, the clearance of LDL appeared to be impaired and a higher proportion of VLDL was metabolized without conversion to LDL. There were significant changes in the flux of particles to the LDL compartment. Total and VLDL TG concentrations were found to be inversely related to rates of insulin-mediated glucose disposal, and HDL cholesterol concentrations were positively related to glucose disposal. These relationships between lipoproteins and insulin action were independent of adiposity and insulin, suggesting that insulin resistance may be involved with diabetes-induced changes in VLDL and HDL. Weight loss was associated with decreases in total and VLDL TG, decreases in total and LDL cholesterol, and improvements in the ratio of HDL to LDL cholesterol. Sulfonylurea therapy was associated with lower total and VLDL TGs and lower LDL cholesterol, but little change in HDL. Substitution of complex carbohydrates for saturated fat in the diet showed consistent and significant decreases in total and LDL cholesterol, no decreases in HDL cholesterol, or elevation of total or VLDL TG. CONCLUSIONS--Studies suggest that there are multiple changes in plasma lipoproteins accompanying NIDDM in Pima Indians, but that many of these may be reversed by current modes of hypoglycemic therapy. PMID- 8422794 TI - Clinical hypertension and its interaction with diabetes among American Indians and Alaska Natives. Estimated rates from ambulatory care data. AB - OBJECTIVE: To estimate the prevalence of clinical hypertension and describe the coexistence with diabetes in American Indian and Alaska Native communities. RESEARCH DESIGN AND METHODS: A cross-sectional study of outpatient visits for hypertension and diabetes over a 1-yr period (1 October 1986 to 30 September 1987) in IHS facilities was conducted. RESULTS: The 1987 estimated age-adjusted prevalence of diagnosed hypertension for this group was 10.9/100 for people > or = 15 yr of age. Thirty-seven percent of diabetic patients were diagnosed with hypertension. The relative risk of hypertension in the diabetic populations compared with the nondiabetic population varied from 4.7 to 7.7 among the different IHS areas. CONCLUSIONS: Despite high rates of diabetes and obesity, hypertension rates were relatively low among American Indians and Alaska Natives when compared with other ethnic groups in the U.S. PMID- 8422795 TI - Diabetes-associated mortality in Native Americans. AB - OBJECTIVE: To describe diabetes-associated mortality among Native Americans. RESEARCH DESIGN AND METHODS: In this population-based study, we analyzed diabetes associated mortality data from the IHS and the NCHS. We also examined diabetes data from the 1986 NMFS. RESULTS: IHS area-specific diabetes mortality rates for 1984-1986 ranged from 10 to 93/100,000, compared with 15/100,000 for the total U.S. population. NCHS data for the same period listed diabetes as the underlying cause of 708 deaths among Native Americans and the contributory cause of 1252 deaths; 63% of the latter deaths were attributable to circulatory diseases. The 1986 NMFS demonstrated that Native American heritage is underreported by 65% on death certificates. Using deaths identified as Native American by NMFS, the age adjusted mortality rate for diabetes as the underlying cause for Native Americans (96/100,000) was 4.3 times that for whites and two times that for blacks. Where diabetes was a contributory cause of death, the mortality rate for Native Americans (264/100,000) was 3.7 times that for whites and 2.4 times that for blacks. CONCLUSIONS: The excessive diabetes-associated mortality among Native Americans is consistent with other indicators of the magnitude of the diabetes problem in this population. Further epidemiological research and expanded diabetes control interventions are needed. PMID- 8422796 TI - A follow-up study of diabetic Oklahoma Indians. Mortality and causes of death. AB - OBJECTIVE: To determine the mortality rates and causes of death for diabetic Oklahoma Indian adults by sex and age. RESEARCH DESIGN AND METHODS: This was a cohort follow-up study with baseline examination between 1972-1980 and the mortality follow-up between 1986 and 1989. Mean follow-up time was 10 +/- 4 yr. A quasi-random sample of 1012 (379 men and 633 women) NIDDM American Indians in Oklahoma was performed. Mean age was 52 yr and duration was 7 yr at baseline. Mean degree of Indian blood was 92% (77% full blood). At follow-up, 548 (54%) were alive, 452 (45%) were decreased, and 12 (1%) could not be traced. RESULTS: Death certificates were obtained and coded (ICD-9) for 439 (97%) of the deceased. Mean annual mortality rates were 4.99% for men and 4.17% for women, with an increasing trend with age for both sexes. Compared with the general population of Oklahoma, the observed/expected ratios for number of deaths were 2.92 for men and 4.09 for women (P < 0.0001). The three leading causes of death were circulatory disease (38%), diabetes (24%), and malignant neoplasms (12%). CONCLUSIONS: There is an excessively high mortality among diabetic Oklahoma Indians compared with the general population in the state and with diabetic patients in other populations. PMID- 8422797 TI - Diabetes mortality among New Mexico's American Indian, Hispanic, and non-Hispanic white populations, 1958-1987. AB - OBJECTIVE: To determine the diabetes-related mortality rates among New Mexico's American Indians, Hispanics, and non-Hispanic whites over a 30-yr period. RESEARCH DESIGN AND METHODS: Death certificates were used to identify diabetes as an underlying cause of death by ethnic group in New Mexico during each 5-yr period from 1958 through 1987. The age-adjusted rates were calculated by ethnic group and sex, and temporal trends were examined. Comparison was made to U.S. white age-adjusted rates during the same time period. RESULTS: Age-adjusted diabetes mortality rates for American Indians and Hispanics increased throughout the 30-yr period, and far exceeded rates for New Mexico non-Hispanic whites and U.S. whites by the 1983-1987 time period. The rates increased most dramatically among the state's American Indians, increasing 550% among women and 249% among men. Hispanic women and men experienced increases of 112 and 140%, respectively. CONCLUSIONS: New Mexico's American Indian and Hispanic populations have higher diabetes mortality rates than non-Hispanic whites, and American Indian mortality rates have risen dramatically over the 30-yr period included in our study. Although the high prevalence of diabetes in American Indians and Hispanics is a major contributor to these rates, other factors may also influence the reported mortality rates. PMID- 8422798 TI - Diabetes and obesity in the offspring of Pima Indian women with diabetes during pregnancy. AB - OBJECTIVE: To review the long-term effects of the diabetic pregnancy on the offspring among the Pima Indians of Arizona. RESEARCH DESIGN AND METHODS: Studies published by the Phoenix Epidemiology and Clinical Research branch of the National Institute of Diabetes and Digestive and Kidney Diseases, since the inception of the longitudinal diabetes studies in 1965 were reviewed. In addition, pertinent studies from other centers, mentioned as references in these publications, were reviewed. As far as possible, all original articles and abstracts on this aspect of the Pima Indian studies were discussed. RESULTS: The offspring of women who had diabetes during pregnancy, on average, were more obese and had higher glucose concentrations and more diabetes than the offspring of women who developed diabetes after pregnancy or who remained nondiabetic. Although no new analyses were attempted, several of the older publications were updated by repeating the analyses on later, expanded data sets. CONCLUSIONS: The diabetic pregnancy, in addition to its effects on the newborn, has effects on the subsequent growth and glucose metabolism of the offspring. These effects are in addition to genetically determined traits. PMID- 8422799 TI - Prevalence of diabetes mellitus in pregnancy among Yup'ik Eskimos, 1987-1988. AB - OBJECTIVE: To evaluate the prevalence of diabetes mellitus in pregnancy in Yup'ik Eskimos. RESEARCH DESIGN AND METHODS: A retrospective review of consecutive birth log data and medical records was conducted. Records were taken from the primary care and tertiary referral facilities in Alaska Area Native Health Service. Some 630 consecutive deliveries were reviewed from 1 March 1987 to 29 February 1988, with 25 excluded for ancestry other than Alaska Native. Another 605 Alaska Native patient charts were reviewed, with 545 Alaska Native patients screened for diabetes in pregnancy. The study population had a mean age of 25.6 yr, mean gravidity of 3.4, mean parity of 1.9, and mean birth weight of 3567 +/- 493 g. RESULTS: Patients were screened with a 50-g glucose oral load with a plasma glucose 1 h later. 156 of 605 (25.7%) patients, with a screen > or = 7.8 mM received a 100-g OGTT. Of 605, 35 (5.8%) patients met O'Sullivan criteria, and 2 of 605 (0.3%) patients met WHO criteria for previous diabetes mellitus, for a total 37 of 605 (6.7% [corrected]) women with diabetes in pregnancy. The subjects who met O'Sullivan criteria had statistically greater mean age (29.9 yr), gravidity (4.9), parity (2.9), and birth weight of their infants (3678 +/- 389 g), compared with women with a screen < 7.8 mM. CONCLUSIONS: The prevalence of diabetes in pregnancy among Yup'ik Eskimos is twice the rate for the U.S. for all races, despite the Yup'ik having the lowest rate of diabetes mellitus among Alaska Natives. This may represent a large number of undiagnosed patients with impaired glucose tolerance, and may reflect the wide-spread dietary and life style changes that have occurred in the Yup'ik in the last 30 yr. The Yup'ik present a unique opportunity to apply prevention techniques in a population with an emerging problem with glucose tolerance. PMID- 8422800 TI - Diabetes mellitus in Tohon O'odham pregnancies. AB - OBJECTIVE: To determine the prevalence of diabetes in Tohono O'odham pregnancies, and to assess the efficacy of early prenatal diabetes screening in populations with high rates of diabetes. RESEARCH DESIGN AND METHODS: All Tohono O'odham women receiving prenatal care at the Sells Service Unit who delivered a viable baby in the 5-yr study period were reviewed. RESULTS: Diabetes was confirmed in 97 of 1854 (5.2%) Tohono O'odham pregnancies delivered during the study period. NIDDM antedated pregnancy in 38 (39%) of the diabetic pregnancies. Diabetes was diagnosed for the first time in 59 (61%) of these diabetic pregnancies. The diagnosis was made at or before the 20th gestational wk in 25 of 59 (42%) gestational diabetic women. These 25 women represented 61% of 41 gestational diabetic women who received prenatal care at or before the 20th gestational week. CONCLUSIONS: Our findings indicate that first trimester diabetes screening is justified in this population and may be appropriate in other populations with high rates of diabetes. PMID- 8422801 TI - Hyperinsulinemia in macrosomic infants of nondiabetic mothers. AB - OBJECTIVE: We tested the hypothesis that macrosomic infants of nondiabetic mothers are more likely to have hyperinsulinemia and increased subcutaneous fat. RESEARCH DESIGN AND METHODS: Plasma insulin concentrations were measured in cord blood from 50 macrosomic infants and 32 normal-sized (control), term infants. All mothers had had a normal 50-g 1-h GCT. Skin-fold measurements of the triceps and subscapular area were done on 44 macrosomic infants with a Halpern caliper. RESULTS: No difference was observed in GCT between mothers of macrosomic (5.8 +/- 1.0 mM) and normal (5.7 mM) infants. The insulin level in macrosomic infants (18.75 +/- 19.08 microU/ml) was significantly higher than in control infants (8.67 +/- 6.64 microU/ml). Macrosomia was a predictor of hyperinsulinemia and vice versa (R2 = 0.26). Maternal height, prepregnancy weight, and weight gain were predictors for macrosomia (R2 = 0.26). No differences were noted in anthropometric measurements between hyperinsulinemic and normoinsulinemic infants. CONCLUSIONS: A subset of macrosomic infants have hyperinsulinemia. Maternal anthropometric factors as well as hyperinsulinemia are correlated with macrosomia. The macrosomia may be causally related to the high insulin levels. PMID- 8422802 TI - Diabetic retinopathy. AB - OBJECTIVE: To review the natural history and treatment indications for diabetic retinopathy. RESEARCH DESIGN AND METHODS: Studies of the natural history of diabetic retinopathy and major clinical trials sponsored by the National Eye Institute were reviewed. RESULTS: Diabetic retinopathy remains a leading cause of vision disability in the U.S. Vision loss results from the consequences of proliferative retinopathy or macular edema. Photocoagulation has been demonstrated to be an effective treatment for these complications of diabetes. CONCLUSIONS: All people with diabetes should be evaluated regularly to determine whether they would benefit from photocoagulation. As a referral guideline, any person with type I diabetes of > 5 yr duration and all people with type II diabetes or any retinopathy should be referred to an ophthalmologist. PMID- 8422804 TI - Periodontal disease. The sixth complication of diabetes mellitus. PMID- 8422803 TI - Factors associated with failure to complete treatment for diabetic retinopathy among Navajo Indians. AB - OBJECTIVE: To determine the proportion of patients with suspected proliferative diabetic retinopathy who did not receive the recommended follow-up ophthalmological evaluation and care, and to examine associations between various patient characteristics and the failure to obtain care. RESEARCH DESIGN AND METHODS: The study cohort included all Navajo Indians identified by a retrospective review of records who had proliferative diabetic retinopathy diagnosed at an Indian Health Service Optometry Clinic between 1 October 1985 and 30 September 1988. Follow-up data were obtained by medical record reviews and by interviews with subjects. RESULTS: Of 69 patients identified, 57 of 61 living patients were interviewed. Twenty-three (40.4%) had failed to obtain recommended follow-up. The RR for incomplete treatment among those without a vehicle in the household compared with those with a vehicle was 1.91 (95% CI 1.32-2.76). Other factors associated with incomplete treatment were female sex and marital status other than currently married. Twelve (21%) patients answered "no" to the question, "Have you been told that diabetes was affecting your eyes?" Eight of 38 (21%) who confirmed that they had been told that diabetes was affecting their eyes responded "no" to the question, "Do you think that diabetes is affecting your eyes?" However, the answers to these questions did not distinguish between patients who obtained or did not obtain recommended care. CONCLUSIONS: Interventions to increase the proportion of Navajo Indians with diabetic retinopathy who receive appropriate ophthalmologic care must address the issue of transportation. PMID- 8422805 TI - Diabetic kidney disease in Pima Indians. AB - OBJECTIVE--To describe the natural history of kidney disease in Pima Indians with NIDDM. RESEARCH DESIGN AND METHODS--Review of previous studies describing diabetic kidney disease in this Native-American population and in other populations. RESULTS--NIDDM is the leading cause of renal failure in Pima Indians, among whom the incidence of ESRD is 23 times that of the general U.S. population. The high incidence of NIDDM and its early onset in the Pima undoubtedly contribute to this difference. The incidence of overt nephropathy and ESRD, as a function of diabetes duration, is at least as high in Pima Indians with NIDDM as that reported in other populations with IDDM. Furthermore, nearly all of the excess mortality associated with NIDDM is found in individuals with overt nephropathy. Mild elevations of UAE, which may be present even shortly after the onset of diabetes, predict the development of overt nephropathy in diabetic Pimas. Additional predictors include high blood pressure, level of glycemia, duration of diabetes, family history of diabetic nephropathy, and type of diabetes treatment. CONCLUSIONS--Diabetic kidney disease is a major cause of morbidity and mortality in Pima Indians. The natural history of diabetic kidney disease in this population is similar, in many ways, to the natural history described in individuals with IDDM. PMID- 8422806 TI - Renal disease among the Eastern Band of Cherokee Indians. AB - OBJECTIVE: To ascertain the incidence and prevalence of ESRD in the Eastern Band of Cherokee in the IHS user population from 1978 to 1988 and to determine what proportion of ESRD and chronic renal failure is attributable to diabetes. RESEARCH DESIGN AND METHODS: Cases were identified from three IHS data sources and from death records obtained from the North Carolina Bureau of Health Statistics. Chronic renal insufficiency was defined as an individual having a serum creatinine of > or = 176.8 microM that remained at that level for a sustained period. An ESRD case was defined as one in which dialysis was required to sustain life, regardless of whether treatment was actually implemented. Data were obtained from chart review and included date of diagnosis, maximum serum creatine level attained, diabetes status, and certain demographic data. RESULTS: Twenty-two (88%) cases of ESRD were attributable to diabetes. The average annual incidence of ESRD during the study period was much higher than that in the U.S. white population and that of Native Americans. The incidence of ESRD caused by diabetes was 2.5 times higher than that reported in the U.S. Native American population affected by diabetes. Degree of Indian inheritance did not appear to be related to prevalence of diabetic renal disease in individuals having diagnosed diabetes. CONCLUSIONS: The proportion of ESRD attributable to diabetes indicates that primary prevention of diabetes may be the best method of preventing ESRD in this population. PMID- 8422807 TI - Diabetic end-stage renal disease among Native Americans. AB - OBJECTIVE: To examine why ESRD has become a major source of morbidity and mortality for Native Americans with diabetes mellitus. RESEARCH DESIGN AND METHODS: Using data from the Medicare ESRD Program, we examined incidence rates for ESRD among Native Americans for the years 1983-1987. RESULTS: During this period, the annual incidence of total ESRD in Native Americans increased by 18%, from 170.5/million to 200.1/million. The incidence of diabetic ESRD increased by 47%, from 80.6/million to 118.2/million. In 1987, the age-adjusted incidence rate of diabetic ESRD was 6.8 times higher in Native Americans than in whites. CONCLUSIONS: Recommendations for the prevention of diabetic ESRD include early identification of renal disease and improved control of hypertension and blood glucose. The magnitude of diabetic ESRD among Native Americans also underscores the need for primary prevention of non-insulin-dependent diabetes mellitus. PMID- 8422808 TI - Epidemiology of lower-extremity amputations in the Indian Health Service, 1982 1987. AB - OBJECTIVE: To describe the LEA experience among IHS diabetic patients. RESEARCH DESIGN AND METHODS: A cross-sectional study of hospital discharges for nontraumatic LEAs from 1982 to 1987 in four IHS areas was conducted. RESULTS: Incidence rates of first LEA in the study period increased with increasing age. Compared with nondiabetic subjects, diabetic patients had increased risks in each age-group, with those between the ages of 15 and 44 yr having a 158-fold increased risk. The average annual age-adjusted incidence rates of all LEAs among diabetic subjects in the Tucson (240.8/10,000) and Phoenix (203.1/10,000) IHS areas were substantially higher than the rates for the U.S. (73.1/10,000), Navajo (74.0/10,000), and the Oklahoma (87.3/10,000) IHS areas. CONCLUSIONS: LEA rates varied in different IHS areas. Reasons for these findings need to be evaluated, but may include IHS area differences in preventive health-care practices or risk factors for LEA. PMID- 8422809 TI - Prevalence and risk factors for diabetes and diabetes-related amputations in American Indians in southern Arizona. AB - OBJECTIVE: To describe the prevalence of NIDDM and LEA using data from a computer based patient data base. RESEARCH DESIGN AND METHODS: Diabetic patients with and without LEA, and nondiabetic patients were identified by computer search. Charts of diabetic patients were reviewed for confirmation of diagnosis of diabetes and diabetes-related amputation. The diabetic and nondiabetic populations were described, and certain risk factors were identified. RESULTS: The overall prevalence of NIDDM in this tribe in 1985-1986 was 18.3/100 adults (> or = 18 yr of age), whereas the prevalence of LEA/100 adults with NIDDM was 10.3%. Females were 1.3 times as likely to have diagnosed diabetes as males (95% CI 1.2-1.4), and males with diabetes were 1.4 times more likely to have had LEA than females with diabetes (95% CI 1.1-1.9). CONCLUSIONS: Automated health-care delivery data base used for this tribe can be used to maintain surveillance for diabetes and amputations in diabetic patients. Effective programs to prevent complications of diabetes, such as LEA, in this tribe are urgently needed. PMID- 8422810 TI - Diabetic complications among American Indians of Washington, Oregon, and Idaho. Prevalence of retinopathy, end-stage renal disease, and amputations. AB - OBJECTIVE: To estimate the prevalence of severe diabetic complications. RESEARCH DESIGN AND METHODS: We surveyed the data systems of the IHS and local care providers to identify all AI/AN people with any of three diabetic complications: retinopathy needing one or more laser treatments, ESRD needing dialysis, and one or more LEAs. We calculated prevalence rates that were sex-age adjusted to the U.S. population for total sample, each community, gender, and culture area. RESULTS: Of the population-based complete sample with diabetes, 1.7% had retinopathy, 1% ESRD, and 4% LEA. Complication rates among the culture areas were not statistically different. The LEA rate among diabetic men (5.8%) was higher than that among women (2.3%). The odds ratio was 2.2 (95% CI 1.2-4.2). One community had an extraordinarily high LEA rate of 16%. CONCLUSIONS: This study showed that IHS can use its data systems to estimate prevalence of diabetic complications and identify population groups with higher prevalence rates. The data led the IHS clinic with the high LEA rate to start a program to prevent amputations. We suggest quality assurance activities based on the methods of this study. PMID- 8422811 TI - The diabetes project at Fort Totten, North Dakota, 1984-1988. AB - OBJECTIVE: To characterize demographic, therapeutic, and complication features of patients in the Fort Totten Diabetes Project and to assess the longitudinal impact of intervention strategies. RESEARCH DESIGN AND METHODS: Individuals (n = 215) of varying American Indian ethnic origin and a quantum of mean age 53 yr (range 14-86 yr), 62% women, with type II diabetes of 9 yr average duration were studied. Annual chart audits of the complete diabetic population were conducted from 1984 to 1988. RESULTS: Obesity is a major problem complicating diabetes management (average BMI 32 +/- 0.9), but BMI does not correlate with glucose control as assessed by total GHb (r = 0.014, NS). Individuals lost an average of 3.7 kg (P < 0.001) during the audit years. Mean GHb declined for the population (11.3-9.7%, P < 0.025) during the interval, but no significant decline was evident when values for individuals were considered. Complication rates for retinopathy, nephropathy, and neuropathy paralleled those rates published for urban populations. CONCLUSIONS: Diabetes mellitus in American Indians living in a rural, economically deprived area can be identified and cared for with success similar to that published for urban populations. PMID- 8422813 TI - The traditional Pima Indian diet. Composition and adaptation for use in a dietary intervention study. AB - OBJECTIVE: To examine changes in the Pima Indian diet composition that may have played a role in the dramatic rise in the incidence of NIDDM among Pima Indians over the last century. RESEARCH DESIGN AND METHODS: We investigated the composition of the foods comparable to those available to the Pima approximately 100 yr ago, with the aim of reproducing this traditional diet as faithfully as possible for a dietary intervention study. An approximation of the traditional diet was ascertained from the ethnohistoric literature and traditional recipes. RESULTS: We estimated that the traditional Pima diet, although seasonably variable, was approximately 70-80% carbohydrate, 8-12% fat, and 12-18% protein. A diet analogous to the traditional Pima diet is largely reproducible with the foods available today. Many native foods are available locally and many commercial products can be substituted when native foods are unavailable. CONCLUSIONS: The Pima Indian diet of the last century was much higher in carbohydrate and lower in fat compared with the modern-day Pima diet. Any changes that this diabetes-prone population can make toward their traditional diet may help to decrease their incidence of diabetes. PMID- 8422812 TI - Insulin therapy and weight change in Native-American NIDDM patients. AB - OBJECTIVE: To determine whether NIDDM patients exposed to insulin therapy in a clinical setting gain weight. RESEARCH DESIGN AND METHODS: This study, an historical cohort chart review, was conducted at the IHS clinic on the Navajo Reservation in Northern Arizona. We studied 27 Native Americans with NIDDM and 102 Native-American nondiabetic control subjects. RESULTS: Insulin therapy consisting of a mean of 105 U/day was associated with a mean weight gain of 3.0 +/- 2.2 kg/yr. When insulin was discontinued or decreased, a mean weight loss of 5.2 +/- 2.7 kg/yr was observed in the same patients. No significant weight gain was noted in 102 nondiabetic control subjects, nor in 20 of 27 insulin-treated patients given oral hypoglycemic agents before initial insulin therapy. CONCLUSIONS: Insulin therapy appeared to be associated with weight gain in this group of NIDDM patients. This suggests that observations for weight gain be undertaken when treating NIDDM patients with insulin, because it may exacerbate the underlying pathophysiology of the disease. PMID- 8422814 TI - Effects of gemfibrozil on triglyceride levels in patients with NIDDM. Hyperlipidemia in Diabetes Investigators. AB - OBJECTIVE: Patients with NIDDM have a two- to fourfold increased risk of macrovascular disease. The constellation of elevated TGs and decreased HDL cholesterol are recognized as risk factors and constitute the major dyslipidemia in NIDDM. We therefore sought to determine if gemfibrozil (600 mg b.i.d.) was effective in correcting the dyslipidemia of NIDDM. RESEARCH DESIGN AND METHODS: After 8 wk of placebo stabilization, 442 patients from 46 study centers were randomized to double-blind treatment, in a designated 2:1 ratio, 295 received gemfibrozil and 147 received placebo for 20 wk. The primary end point was plasma TG; secondary end points were TC, LDL cholesterol, VLDL cholesterol, HDL cholesterol, and HbA1c. No baseline differences were noted between groups in sex, age, weight, type of diabetic therapy, fasting plasma levels of TGs, HbA1c, or C peptide. About two-thirds received oral hypoglycemic drugs, one-third insulin. RESULTS: TG fell 26.4% in the gemfibrozil group and rose 7.4% in the placebo group (P < 0.023), by an intent-to-treat analysis. When patients who were noncompliant or with inadequate data were excluded, similar results were found--a 30.4% fall with gemfibrozil and a 4.8% increase with placebo (P < 0.0001). TG levels fell within 4 wk and remained low for 20 wk (P < 0.001). Mean HDL cholesterol rose by 4 wk and increased further at 12 wk (8-12%), P < 0.0001. TC fell. We observed a significant rise in LDL cholesterol in both gemfibrozil- and placebo-treated groups, with no significant differences between these groups. Changes in HbA1c were similar in gemfibrozil and placebo groups. No differences were observed in responses in groups treated with insulin and or oral hypoglycemic drugs. Overall AEs that were clinically important occurred in 6.1% in the gemfibrozil group vs. 2.0% in the placebo group (NS). CONCLUSIONS: We conclude that gemfibrozil is an effective and safe agent in combating the dyslipidemia of NIDDM, irrespective of type of diabetic therapy. PMID- 8422815 TI - Improving diabetes care for American Indians. AB - In 1986, a diabetes control program was implemented in the Billings area of the IHS. Baseline health-care practices in the program were described using a structured audit. The program included adoption of the IHS Minimum Standards of Care for diabetes, technical assistance, and professional and patient education. A second audit was performed in 1988. Care practices improved significantly for all facilities in 7 of 10 parameters measured. Facilities that implemented key program activities showed more overall improvement in screening practices, education, and immunization than those that did not organize diabetes care. Factors associated with improved care practices include establishment of a coordinated, multidisciplinary diabetes team with regular meetings, acceptance of standards of care by the medical staff, use of flow sheets by multiple providers, and diabetes-related professional and patient education sessions. PMID- 8422816 TI - Prevention and early treatment of NIDDM. PMID- 8422817 TI - Treating diabetes and its complications. PMID- 8422818 TI - Diabetes in American Indians. Reflections and future directions. PMID- 8422819 TI - Fetal pancreas transplantation for treatment of IDDM patients. PMID- 8422820 TI - Diabetes and the World Health Organization. Progress towards prevention and control. PMID- 8422821 TI - Need to assess readability of written materials for diabetes education curricula. PMID- 8422822 TI - A joint venture with the commonwealth of independent states. PMID- 8422823 TI - Diabetes patients and their physicians--all eager for knowledge from the West. PMID- 8422824 TI - Effect of meal frequency on blood glucose, insulin, and free fatty acids in NIDDM subjects. AB - OBJECTIVE: We studied the effects of meal frequency on blood glucose, serum insulin, and FFAs in 12 NIDDM subjects. RESEARCH DESIGN AND METHODS: Subjects were assigned in random order to two 8-hr observation periods after an overnight fast. They received isocaloric diets with similar composition either as six small or as two large meals. At the end of each study period, an IVGTT was given. RESULTS: Two large meals induced an 84% greater maximum amplitude of glucose excursions than six small meals (6.1 +/- 0.5 vs. 3.3 +/- 0.5 mM, P < 0.005) and higher insulin responses (P < 0.03). The Kg response to an IVGTT did not differ in the two situations. The average FFA level was lowest in response to frequent meals (P < 0.02). CONCLUSIONS: A higher meal frequency acutely subdues glucose excursions and reduces insulin and FFA levels during the daytime in older NIDDM subjects. PMID- 8422825 TI - Is the traditional alcohol wipe necessary before an insulin injection? Dogma disputed. PMID- 8422826 TI - Effects of pravastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, on glucose tolerance in patients with essential hypertension. PMID- 8422827 TI - Absence of thyroid disease in Chinese children with IDDM. PMID- 8422828 TI - Relevance of HLA-DR phenotype and breast-feeding status in age heterogeneity at IDDM onset. PMID- 8422829 TI - Severe sensory motor polyneuropathy at the onset of diabetes mellitus. Total remission with testosterone therapy following recognition of associated hypogonadism. PMID- 8422830 TI - Estimated creatinine clearance is not an accurate index of glomerular filtration rate in normoalbuminuric diabetic patients. PMID- 8422831 TI - Can islet cell antibodies predict IDDM in the general population? AB - OBJECTIVE: To evaluate the likely prognostic significance of ICAs in children with no family history of IDDM. RESEARCH DESIGN AND METHODS: We examined the prevalence of ICAs in 2925 English schoolchildren aged 9-13 yr and in 274 age matched siblings of children with diabetes from the same region, and we compared the estimated risk of progression to diabetes within 10 yr in the two groups. RESULTS: ICAs were present at levels > or = 4 JDF U in 2.8% of schoolchildren and 6.6% of siblings and at > or = 20 JDF U in 0.8% of schoolchildren and 2.2% of siblings. Although ICAs are only 2-3 times more prevalent in siblings than schoolchildren, the estimated cumulative risk that siblings will progress to diabetes by age 21 is 13 times greater (2.8 vs. 0.21%). CONCLUSIONS: ICAs are unexpectedly prevalent in English schoolchildren, but only a small minority, with this evidence of immune activation directed against islet cells, will progress to diabetes. Although ICAs alone have limited predictive value in the general population, combining two or more predictive tests in series could achieve a level of prediction equivalent to that now obtained in first-degree relatives. PMID- 8422832 TI - Lipid tolerance testing in pregnancy. AB - OBJECTIVE: Macrosomia in the infant of the well-controlled diabetic mother suggests that a substrate other than glucose may influence fetal growth. We hypothesized that GDM alters lipid homeostasis during pregnancy. Our objective was to determine if an LTT could assist in identification of lipid abnormalities in the GDM individual. RESEARCH DESIGNS AND METHODS: The LTT consisted of bolus infusion of 1.0 mg/kg 10% Intralipid (Cutter Vitrum, Berkeley, CA) followed by measurements of plasma glucose, insulin, glycerol, total triglycerides, and triglyceride fatty acids (18:1 and 18:2), total FFAs, and total phospholipids for 30 min before and 90 min after the bolus. The study groups were composed of 8 nonpregnant, nondiabetic subjects, 8 pregnant, nondiabetic subjects, 8 GDM patients receiving insulin, and 8 GDM patients who were diet controlled. RESULTS: Plasma glucose and plasma insulin concentrations did not change significantly after the bolus. No significant difference was noted in the K2 for glycerol of the nonpregnant, nondiabetic group compared with the pregnant groups. The K2 for total triglycerides, plasma triglyceride 18:1, and plasma triglyceride 18:2 indicated increased rates of disappearance of these substrates for the nonpregnant nondiabetic group compared with all pregnant groups. No significant differences were observed among the pregnant groups for any of these parameters. CONCLUSIONS: Pregnancy may be associated with a decreased rate of triglyceride lipolysis compared with nonpregnancy. No differences in lipid metabolism were noted among normal pregnant and relatively well-controlled GDM patients. PMID- 8422833 TI - Elevated serum sialic acid concentration in NIDDM and its relationship to blood pressure and retinopathy. AB - OBJECTIVE: In view of the possible link between serum sialic acid and cardiovascular disease in the general population, we investigated whether serum total and lipid-associated sialic concentrations are elevated in NIDDM patients compared with normal subjects. We also investigated how sialic acid levels relate to glycemic control, blood pressure, microalbuminuria, retinopathy, and serum lipid levels. RESEARCH DESIGN AND METHODS: We selected 20 NIDDM patients at random and matched them for age and sex with 20 normal subjects. The patients also had a similar BMI as the control subjects. A first morning blood sample was taken for sialic acid, glucose, fructosamine, and lipid analysis, as was a first morning urine sample for assessment of microalbuminuria. Retinopathy was assessed by fundoscopy. RESULTS: Both total and lipid-associated sialic acid levels were elevated in the NIDDM patients compared with control subjects (mean +/- SD, total: 0.74 +/- 0.11 vs. 0.60 +/- 0.22 g/L, P < 0.02; lipid-associated: 0.18 +/- 0.04 vs 0.12 +/- 0.04 g/L, P < 0.001). Total serum sialic acid was correlated with systolic blood pressure (r = 0.58, P < 0.01) and diastolic blood pressure (r = 0.58, P < 0.02). There was no significant relationship of total sialic acid with age, duration of diabetes, BMI, microalbuminuria, serum triglyceride, blood glucose, or serum fructosamine. A relationship of lipid-associated sialic acid levels and systolic blood pressure did not reach significance (P = 0.09). In 9 patients with background retinopathy with or without maculopathy, the total serum sialic acid concentration was higher than in those without retinopathy (0.81 +/- 0.09 vs. 0.69 +/- 0.10 g/L, P < 0.008). Lipid-associated sialic acid levels were similar in those with and without retinopathy. (The conversion factor for standard units to SI units is 1 gL = 3.2 mM.) CONCLUSIONS: Total serum sialic acid levels were significantly elevated in a relatively small group of NIDDM patients and were correlated with hypertension and retinopathy. A larger study of circulating sialic acid concentrations as a risk factor for the development or marker of diabetic angiopathy is therefore justified. PMID- 8422834 TI - Autonomic function in neuropathic diabetic patients with foot ulceration. AB - OBJECTIVE: The aim of this study was to compare peripheral autonomic function in neuropathic diabetic subjects with and without foot ulceration. RESEARCH DESIGN AND METHODS: Measurements were made on 57 diabetic subjects; 35 subjects had evidence of peripheral neuropathy, 14 of these had a history of foot ulceration, and 22 subjects had no evidence of peripheral neuropathy. No patient had peripheral vascular disease. Measurements were made of motor and sensory nerve conduction. Autonomic function was investigated by using standard cardiovascular reflex tests and by measuring blood flow variability in the foot. The vasoconstrictor responses to deep breathing and body cooling were measured by using venous occlusion plethysmography. RESULTS: Peripheral sympathetic function was significantly worse in the DU group. The vasoconstrictor response to deep breathing in the DU group was significantly smaller than the response in the DN group (15.3 +/- 2.7 vs. 38 +/- 4%, P < 0.001). The response to body cooling in the DU group was significantly smaller than the response in the DN group (6.2 +/- 3.1 vs. 20.8 +/- 3.5%, P < 0.01). Tests of cardiac autonomic function and measurements of motor and sensory nerve conduction were similar in both neuropathic groups. CONCLUSIONS: Peripheral autonomic neuropathy is associated with the development of foot ulceration in diabetic subjects. PMID- 8422835 TI - Prevalence of NIDDM and impaired glucose tolerance in aborigines and Malays in Malaysia and their relationship to sociodemographic, health, and nutritional factors. AB - OBJECTIVE: To determine the prevalence of diabetes mellitus and IGT in different ethnic groups living in the same physical environment and to find their relationship to nutritional status and dietary intake. RESEARCH DESIGN AND METHODS: The study was conducted among Malays and Orang Asli in six rural and urban locations in Malaysia. OGTTs were performed on 706 adult subjects > or = 18 yr of age. WHO criteria were used for diagnosing diabetes mellitus and IGT. RESULTS: The overall prevalence of diabetes mellitus and IGT among Orang Asli was 0.3 and 4.4% compared with 4.7 and 11.3%, respectively, among Malays. This increased prevalence of glucose intolerance among Malays was associated with higher levels of social development. Among rural Malays, the crude prevalence of diabetes in a traditional village was 2.8% and in the land scheme was 6.7%, whereas urban Malays had a prevalence of 8.2%. In contrast, the prevalence of IGT (10.5-14.8%) was higher among rural Malays, compared with 9.6% among urban Malays. Ethnic group, > or = 40 yr of age, an income > M$250, fewer daily activity, and obesity were associated with a higher prevalence of diabetes. CONCLUSIONS: Diabetes mellitus and IGT, which were more common among Malays than Orang Asli, were associated with more affluent life-styles and modernization. PMID- 8422836 TI - C-peptide in NIDDM. Follow-up for 4-6 yr. AB - OBJECTIVE: To study whether fasting and 1-h postbreakfast C-peptide levels in NIDDM stabilize with time in individual patients. RESEARCH DESIGN AND METHODS: Within the period of 4-6 yr, 49 NIDDM patients had repeated tests of fasting and 1-h postprandial levels of plasma glucose and C-peptide with the aim of determining their individual qualitative patterns. Throughout the follow-up period, 13 patients were treated with insulin, 21 with oral sulfonylureas, and 15 were switched from oral drugs to insulin, with the tests done in both treatment periods. RESULTS: The group as a whole demonstrated no changes in mean fasting or postprandial C-peptide within 4-6 yr of observation, irrespective of the mode of therapy or its changes. Glycemic and C-peptide response to breakfast was qualitatively typical for each patient with the correlation between plasma glucose and C-peptide. However, the response was vastly different from patient to patient, and the cross-sectional data showed no correlation between postprandial changes in glycemia and C-peptide, nor between glycemic response to breakfast and fasting plasma glucose or C-peptide. In spite of high fasting glycemia, 25% of the patients showed remarkable tolerance to breakfast with only small increases in plasma glucose. In many other patients, however, in spite of similar increase in C-peptide, plasma glucose rose sharply after the meal. CONCLUSIONS: In our group, no deterioration of the insulin secretory function was observed within 4-6 yr of follow-up. Qualitative patterns of the glycemic and C-peptide responses to breakfast were typical for each patient but vastly different between patients. We see in NIDDM a syndrome with few common characteristics and recommend further work for its subclassification into forms with different pathogenesis. PMID- 8422837 TI - Mortality of Mexican Americans with NIDDM. Retinopathy and other predictors in Starr County, Texas. AB - OBJECTIVE: To determine the rate and risk factors of mortality in a cohort of Mexican Americans with NIDDM. RESEARCH DESIGN AND METHODS: A cohort of 353 Mexican Americans with NIDDM were identified between 1981 and 1986. All individuals underwent extensive evaluations that included physical, historical, ophthalmological, and laboratory assessments. This cohort was followed prospectively for a mean of 8 yr. Follow-up included mortality surveillance, death certificate extraction, and a combination of annual and intermediate examinations. RESULTS: The cohort experienced 67 mortality events. One-third of all deaths were premature < 65 yr of age) and most often were attributed to diseases of the heart (60.0%). In no case was diabetes listed as the cause of death, although it was listed as a contributing cause in 25.5% of cases. Men had a higher mortality rate than women. In both sexes, baseline retinopathy was identified as an important predictor of subsequent mortality. Mortality was significantly elevated in those with nonproliferative retinopathy and even further elevated in those with proliferative disease (relative risks of > or = 4 for proliferative disease). CONCLUSIONS: Mexican Americans with NIDDM are experiencing premature and excessive mortality compared with the general population. The results clearly link microvascular complications with macrovascular disease, but this link is not explained by a more untoward profile of traditional cardiovascular risk factors. Retinopathy appears to serve as an important monitor of the progression of diabetes and when identified would warrant aggressive action to inhibit or slow the processes leading to subsequent mortality. PMID- 8422838 TI - Glucose intolerance in Colombia. A population-based survey in an urban community. AB - OBJECTIVE: To determine the prevalence of diabetes and its relationship to age and obesity in an urban community in Colombia. RESEARCH DESIGN AND METHODS: A cluster sample of 670 adults > or = 30 yr of age was selected from the city of Santafe de Bogota. Classification of diabetes and IGT was according to WHO criteria. RESULTS: Response to the survey, conducted in 1988-1989, was 71% for men and 84% for women. Prevalence of diabetes was 7% in both sexes. Prevalence of IGT was 5% in men and 7% in women. Age-standardized prevalence of diabetes in the 30- to 64-yr age range was comparable with that reported in urban Brazilians and rural Hispanics in the U.S.. Prevalence was higher than in the white population of the U.S. but lower than in several urban U.S. Hispanic communities. Some 40% of men and 30% of women with diabetes were unaware of their condition before the survey, but all those < 50 yr of age were diagnosed previously. Glucose intolerance was associated with high BMI in men and with advancing age in both sexes. CONCLUSIONS: Glucose intolerance is common in this community and will likely increase in frequency in Colombians with further urbanization and population aging. PMID- 8422839 TI - Improvement of diabetic control and acceptability of a three-injection insulin regimen in diabetic adolescents. A multicenter controlled study. AB - OBJECTIVE: To compare the effectiveness and acceptability of a three-injection insulin regimen with the conventional two-injection therapy in an unselected population of diabetic adolescents. RESEARCH DESIGN AND METHODS: Some 205 patients aged 10-18 yr with IDDM, who were previously treated with two daily insulin injections, were included without any selection into a randomized trial. They were either switched to three (regular prebreakfast, regular prelunch, and [regular+ultralente] predinner) or remained on two ([regular+intermediary] prebreakfast and predinner) subcutaneous injections. They were evaluated after 1 yr of treatment. The major criteria of outcome of efficacy were the concentration of GHb, the frequency of severe hypoglycemia and DKA, and body weight. RESULTS: Of the patients, 82% accepted the three-injection regimen, and 83% accepted the two-injection regimen. At entry into the trial, no significant differences appeared between the two treatment groups nor among patients refusing the allocated regimen. Significant explanatory variables predicting initial diabetes control were duration of disease and adherence to diet. GHb, decreased from 9.8 +/- 0.1 to 9.3 +/- 0.2% (P < 0.05) in the three-injection group, whereas it increased from 9.5 +/- 0.3 to 9.8 +/- 0.3% (P < 0.05) in the two-injection group, resulting in a modest (0.75%) but significant difference (P < 0.05) between GHb change in the two groups. The difference reached 1.4% (P < 0.0002) in patients with GHb > 11.2% at entry. The frequency of hypoglycemia and DKA was similar in the two groups. None of the parameters known to potentially influence glycemic control changed during the trial, and, therefore, the improvement of GHb could be attributed to the pattern of daily insulin distribution per se. CONCLUSIONS: In the general diabetic adolescent population, the efficacy of a three-injection regimen is somewhat superior to that of a conventional two-injection regimen, particularly in patients previously poorly controlled. The acceptability of this regimen being excellent, its increased use should be considered in this age group. PMID- 8422840 TI - Cystic fibrosis. AB - Cystic fibrosis, an autosomal recessive disorder, is the most common genetic disease of Caucasians. One in 25 Caucasians are carriers of the gene. The gene is found far less commonly in other races. There are over 230 different alleles of the gene, located on the 7th chromosome. The gene encodes for a membrane protein that functions as an ion channel. The survival of cystic fibrosis patients has been gradually increasing, with a mean survival in 1990 of 28 years. If the current trend of improved survival continues, it is estimated that half of cystic fibrosis patients will be over 18 years old by 1996. Disease is found in many organs including the lungs, sinuses, pancreas, gastrointestinal tract, hepatobiliary system, sweat glands and reproductive tract. The majority of patients die of pulmonary disease. The airways become chronically colonized with bacteria that cannot be eradicated, leading to bronchitis, bronchiectasis, and finally, pulmonary fibrosis with respiratory failure. The pulmonary disease may be complicated by massive hemoptysis and pneumothorax. Patient survival rates have increased because of antibiotic therapy and improved nutrition with pancreatic enzyme replacements. New treatments for the pulmonary disease are under clinical trial and include antiproteases, amiloride, a sodium channel blocker, and DNase. The insertion of the normal cystic fibrosis allele into an animal model using a modified adenovirus with effective transcription suggests that gene therapy may be possible in the future, but safety and technical problems have to be addressed. PMID- 8422841 TI - Calcium-dependent regulation of genetically determined spike and waves by the reticular thalamic nucleus of rats. AB - The pathophysiologic role of the reticular thalamic nucleus (RT) in rat generalized nonconvulsive epilepsy was investigated in the selected strain GAERS (genetic absence epilepsy rats from Strasbourg). After the RT was lesioned by the excitotoxic agent ibotenic acid stereotaxically injected in previously callosotomized rats, a disruption of ipsilateral spike and wave discharges (SWD) was observed in freely moving animals. In a second group of animals Cd2+ (0.5-1.5 microliter, 1 mM), which is known to block Ca2+ and Ca(2+)-dependent K+ conductances (gK+ (Ca2+)), was injected into the thalamus. Cd2+ reversibly suppressed ipsilateral SWD when injected in RT, whereas it slightly reduced SWD expression when injected in the ventrobasal (VB) complex. The difference was highly significant. We conclude that Ca(2+)-dependent oscillatory properties of the RT are critical for expression of genetically determined SWD in GAERS. PMID- 8422842 TI - Hypnic tonic postural seizures in healthy children provide evidence for a partial epileptic syndrome of frontal lobe origin. AB - In the pediatric age period, three idiopathic partial epileptic syndromes are recognized: benign childhood epilepsy with centrotemporal spikes, childhood epilepsy with occipital paroxysms, and primary reading epilepsy. All other partial epilepsies are considered cryptogenic, and no other idiopathic partial epilepsies have been recognized. We observed 10 children with tonic partial postural seizures, mainly hypnic, coinciding with a recognizable ictal epileptiform EEG pattern. The children all had normal neuropsychological development before and after seizure onset. The seizures were tractable in all. Onset was in the early pediatric age period; no other type of seizure was recognized. The occurrence of a family history of epilepsy was high. The seizure pattern was typical of supplementary motor area origin. Because of the normal neuropsychological status, high familial incidence of epilepsy, and benign course of this partial epilepsy, we believe it should be considered idiopathic, using the criteria of the International Classification of Epilepsies and Epileptic Syndromes. We therefore suggest the existence of a partial idiopathic epileptic syndrome having onset with seizures of frontal lobe origin. PMID- 8422843 TI - Electroclinical seizures in Lennox-Gastaut syndrome. AB - We analyzed electroclinical seizures observed by long video split-screen recording in 21 patients with Lennox-Gastaut syndrome (LGS). All patients had atypical absence seizures, 18 (81%) had tonic seizures, and 4 (21%) had myoclonic atonic seizures. Tonic seizures were axial with flexion or extension of the head or trunk, or global with generalized tonic spasm mimicking infantile spasm, or involved the eyeballs only (either brief, with upward deviation of eyeballs or long, with oscillatory nystagmus). EEG showed either a bilateral 10-13-Hz rhythm or generalized synchronous spike wave at 3 Hz. Myoclonic-atonic seizures involving limbs, trunk, or neck were either brief or massive; the discharges were 2-3.5-Hz spike wave. Atypical absence seizures evolved gradually, terminated abruptly, and manifested alone or with subtle motor activity or oral automatism. EEG discharges were variable and of different types: (a) Diffuse irregular spike wave at 2-2.5 Hz with or without fragmentation (consciousness was regained during fragmentation or when spike wave discharges were < 2 Hz), (b) irregular diffuse fast activity at 10-13 Hz, or (c) a combination of fast spike wave or sharp waves of increasing amplitude followed by synchronous spike wave discharges at 3 Hz. PMID- 8422844 TI - Primary generalized seizure disorder: correlation of epileptiform discharges with seizure frequency. AB - The role of routine follow-up EEG in assessment of severity of a seizure disorder is not known. This retrospective chart review study of patients with primary generalized seizure disorders explored the relation between parameters of epileptiform discharge bursts on EEG and number of seizures experienced in the preceding months. Eighty-seven such patients were found, each with one to seven EEGs and corresponding clinical records, comprising a total of 201 clinicoelectrographic epochs. The 56 chronically attending patients (more than two attendances) differed from the 31 nonchronic attenders in receiving more antiepileptic drugs (AEDs) and having fewer EEG polyspikes; otherwise, the two groups were similar and were analyzed together. We noted a strong relation between number and maximum length of epileptiform bursts in the resting EEG record and number of absence seizures reported in the months preceding the EEG. Age did not affect this relation. After stepdown regression analysis, only the maximum length of epileptiform bursts remained significantly related to the reported number of absence seizures. These findings may prove useful in assessing clinical progress, e.g., in patients whose ability to estimate numbers of attacks is suspect. Neither number nor maximum length of epileptiform bursts in the resting EEG record had a significant relation with the number of generalized tonic-clonic seizures. Presence of polyspikes on resting or "activated" EEG had no added predictive value. The presence of epileptiform bursts after activation by sleep, hyperventilation, or photic stimulation was not associated with an increased likelihood of having had seizures. PMID- 8422845 TI - Episodic psychic symptoms in the general population. AB - The frequency of some episodic psychic symptoms (dysmnesic, perceptual, and experiential) was determined in a 2,500-subject general population sample. Correlations with some risk factors eventually associated with nervous system dysfunctions (seizure history, head injury, car accident, hospitalization, febrile illness, and birth injury) were calculated. Subjects with one or several risk factors were more likely to report episodic psychic phenomena in daily life. Significant correlations of episodic psychic phenomena with sleep disorders, headache, allergies, and a history of learning disabilities were observed. We propose that some subclinical dysfunctions can be associated with the appearance of episodic psychic phenomena in otherwise normal subjects. PMID- 8422846 TI - Risk factors for developing seizures after a stroke. AB - We evaluated development of seizures in 219 consecutive patients who had ischemic or hemorrhagic stroke. Subjects with transitory ischemic attacks, subarachnoid, subdural, and epidural hemorrhages or those with previous history of epilepsy were excluded. Mean follow-up time was 11.5 months (range 1-72 months). Twenty two of 219 stroke patients (10.04%) had seizures. Twelve (54.55%) were of early onset (< 1 month after the stroke), and 10 (45.45%) were of late onset. No statistically significant differences were evident between the early- and late onset seizure group in comparisons of type of stroke, localization, and size of the lesion. Six of 22 patients (27%) had seizure recurrence. Seizures developed in (a) 13 of 183 patients with ischemic stroke (7.1%) and 9 of 36 patients with hemorrhagic stroke (25%) (p = 0.01); (b) 16 of 93 patients with cortical lesions (17%) and 6 of 126 patients with subcortical lesions (4.7%) (p = 0.01); and (c) 14 of 66 patients with a lesion comprising more than one lobe (21.2%) and 8 of 153 patients with a lesion comprising less than one lobe (5.2%) (p < 0.01). We conclude that patients with hemorrhagic stroke, cortical lesions, and lesions involving more than one lobe are at higher risk of developing seizures. PMID- 8422847 TI - Phenytoin monitoring in status epilepticus in infants and children. AB - Two successive protocols of phenytoin (PHT) plasma concentration monitoring were tested in 60 children with status epilepticus (SE). In each protocol, a loading dose of 15 mg/kg was injected and followed by three injections during the first 24 h. Clinical evaluation was performed at the end of the study by grouping patients into three classes according to seizure frequency during treatment: complete effect, partial effect, and no effect. In protocol 1, a complementary dose at the fourth hour was adjusted from individual plasma concentrations. Plasma concentrations at the 40th hour were within the therapeutic range in the 19 patients with complete effect (CE, mean 19 mg/L) and in the 5 patients with no effect (NE, mean 23 mg/L) whereas in the 11 patients with partial effect (PE), plasma concentrations were higher (mean 31 mg/L). In protocol 2, we added monitoring of the doses injected at the 16th and 24th hours to prevent the increase in PHT concentration noted in protocol 1. All patients but 1 were classified as either CE (13 patients) or NE (4 patients). In NE patients, average plasma concentration at the 40th hour (mean 10.5 mg/L) was lower than in CE patients (mean 15.7 mg/L). In both protocols, the NE patients were the youngest. In SE, PHT doses should be adjusted according to plasma concentrations to avoid overdosage and paradoxical inefficacy. Younger children had lower concentrations and appeared to respond less well than older children, but the etiology of SE may also play an important role. PMID- 8422848 TI - Submandibular salivary gland hypertrophy induced by phenytoin. AB - A 16-year-old boy with epilepsy developed hypertrophy of the submandibular salivary glands, with high phenytoin (PHT) serum levels. The submandibular salivary glands became normal in 12 days after discontinuation of PHT. Other causes of salivary gland hypertrophy were excluded and we suggest that the hypertrophy was due to PHT. PMID- 8422849 TI - Effects of carbamazepine and phenytoin on EEG and memory in healthy adults. AB - Using a randomized, double-blind, cross-over design, we investigated the effects of carbamazepine (CBZ) and phenytoin (PHT) on memory and spectral EEG components in 15 healthy adults. Each subject was treated with each drug for 1 month, separated by a 1-month washout. Evaluations were conducted at baseline, at the end of each treatment month, and 1 month after the last treatment phase. EEG was collected during an eyes-closed resting condition and a verbal memory activation task. Spectral analysis of the EEG in the nondrug conditions showed that the memory task significantly reduced theta components and increased delta components. As compared with nondrug conditions, the antiepileptic drugs (AEDs) significantly impaired memory performance and produced mild EEG slowing. Memory performance did not differ statistically between the AEDs, but minor differences in spectral EEG components were noted. The results suggest that differences in the cognitive and EEG effects of CBZ and PHT are not clinically significant. PMID- 8422850 TI - Comparative pharmacokinetic study of chewable and conventional carbamazepine in children. AB - Fifteen children (7 boys and 8 girls) with generalized tonic-clonic seizures (GTCS) and partial seizures with elementary or complex symptomatology, treated with carbamazepine (CBZ) alone (n = 7) or in combination with either phenobarbital (PB, n = 6) or clobazam (CLB, n = 2) given for at least 3 months at stable individualized doses and regimens, entered an open, within-patient, change over study of consecutive periods, each lasting 2 weeks. During period 1, conventional CBZ was given; during period 2, a chewable CBZ formulation was substituted for conventional CBZ and given at the same total daily dosage with the same schedule as in period 1. Blood samples for measuring plasma concentration of both total CBZ and CBZ-10,11 epoxide (CBZ-E) were taken on the last day of each period. No significant difference between the two periods was noted in the mean +/- SD of Cmax, Css mean, and area under the curve (AUC) of total CBZ and CBZ-E. The two different CBZ formulations, administered at the same total daily dosage, can be considered bioequivalent. PMID- 8422851 TI - Clarithromycin-carbamazepine interaction: a case report. AB - We report a clinically relevant interaction between a new macrolide antibiotic, clarithromycin, and carbamazepine (CBZ). In a patient receiving CBZ monotherapy, 10-day antibiotic treatment increased CBZ concentration despite concomitant CBZ dose reduction and doubled the CBZ concentration/dose ratio. Concentration of the CBZ epoxide (CBZ-E) metabolite was reduced, suggesting that the interaction occurs at a metabolic level. PMID- 8422852 TI - Cross-reactive skin eruption with both carbamazepine and oxcarbazepine. AB - Oxcarbazepine (OCBZ), a 10-keto derivative of carbamazepine (CBZ) has been reported to have a similar range of efficacy and fewer unwanted effects than CBZ since it is a prodrug for the monohydroxy derivative (MHD). A cross-reactivity of only 1 in 4 has been reported between OCBZ and CBZ. For these reasons, we tried OCBZ with 3 consecutive patients with poorly controlled epilepsy who had had a therapeutic response to CBZ but in whom CBZ was discontinued because of serious skin reaction. Each patient had a similar skin response after exposure to only 600-900 mg OCBZ, which suggests a need to practice caution when substituting OCBZ for CBZ in patients known to have serious skin reaction to CBZ. PMID- 8422853 TI - Carbamazepine and lamotrigine in healthy volunteers: relevance to early tolerance and clinical trial dosage. AB - This study was conducted to examine the effects of acute doses of lamotrigine (LTG) and carbamazepine (CBZ) in healthy subjects and determine whether the low tendency to impairment with LTG observed in animals applied to humans. Twelve healthy men participated in a placebo-controlled, balanced, double-blind comparison of the drugs on a series of psychomotor, autonomic, sensory, and subjective variables. Variables were analyzed by analysis of variance, and p < 0.05 was considered significant. Adaptive tracking and body sway were impaired by CBZ 600 mg. CBZ 400 and 600 mg impaired smooth pursuit eye movements and also reduced mean peak saccadic velocity. No differences from placebo occurred after LTG. CBZ 600 mg increased heart rate (HR), but no drug-related changes were noted in pupil size, salivary secretion, visual near point, or subjective effects. During the controlled study, mean plasma CBZ concentrations at 2 and 6.5 h after the 600-mg dose were 5.28 and 5.36 micrograms/ml; after LTG 300 mg, they were 3.16 and 3.00 micrograms/ml. Increased CBZ saliva concentrations were significantly associated (p < 0.01) with impaired adaptive tracking, smooth and saccadic eye movements and increased HR, and plasma concentrations were associated with impaired eye movements and body sway. PMID- 8422854 TI - Acute chemical pancreatitis associated with carbamazepine intoxication. AB - A 5-year-old mentally retarded child developed laboratory evidence of pancreatitis during accidental acute carbamazepine (CBZ) intoxication. He had been seizure-free with CBZ for 4 years for a seizure disorder with no obvious toxicity. CBZ had been discontinued 5 months before he was admitted to the hospital. After he accidentally ingested a CBZ overdose, he was found vomiting and lethargic. Serum amylase and lipase levels were increased for several days. With supportive treatment and no CBZ, he recovered and serum amylase and lipase levels returned to normal. No other causes of pancreatitis were identified. Therefore, most likely the chemical pancreatitis was associated with the acute CBZ intoxication. PMID- 8422855 TI - Valproate-associated pancreatitis. AB - To assess the clinical characteristics of valproate (VPA)-associated pancreatitis, information from three sources was gathered: (a) a survey among 507 physicians with a special interest in treatment of epilepsy, (b) a review of the authors' patient population, and (c) a review of the literature. Of 366 physicians answering the survey, 53 (14.5%) reported a case of pancreatitis. Thirty-nine cases were available for review (24 from the medical literature, 12 from the survey, and 3 from the authors). Pancreatitis appeared to be more frequent in young persons (mean age 16.4 years) but may occur at any age. The highest risk appears to exist during the first months of treatment: 43.8% of the cases developed during the first 3 months, and 68.8% developed during the first year. Seventy-six percent of patients were receiving polytherapy, and 41% had some form of associated chronic encephalopathy. In most patients, the reaction was rapidly reversible when VPA was discontinued. It was severe in 6 patients, with 3 deaths reported. Rechallenge with VPA was attempted in 9 patients, with a high incidence of relapses. Asymptomatic elevation of serum amylase in patients receiving VPA was reported by 40 (10.9%) of the physicians surveyed. Awareness of the problem and early discontinuation of VPA may be effective in preventing serious reactions. PMID- 8422856 TI - Recovery of susceptibility to audiogenic seizure in mice. AB - The applicability of the auditory fatigue and anoxia hypotheses for the refractory period observed after sound-induced seizure was examined in SJL/J mice. Duration of auditory stimulation was varied independent of attainment of clonic seizure activity by use of an acoustic interruption technique. Duration of the recovery period affected the pattern of preconvulsive running. Furthermore, the motor asymmetries exhibited during clonus remained consistent across tests. Because duration of acoustic stimulation and attainment of clonus did not affect recovery rate, we conclude that neither the auditory fatigue nor the anoxia hypothesis provides a complete account for the refractory period after audiogenic seizure. We suggest that an inhibitory process, activated before clonus occurs and perhaps linked to depletion of excitatory amino acids in the inferior colliculus, may also be involved. PMID- 8422857 TI - Carnitine disposition before and during valproate therapy in patients with epilepsy. AB - Free and total carnitine and acylcarnitine in plasma and urine samples was measured in 22 epileptic patients before and after 15 and 45 days of valproate (VPA) therapy and in 16 healthy volunteers on a single occasion. Carnitine plasma concentration and renal excretion observed in epileptic patients before VPA therapy did not differ from control values. After VPA was started, free and total plasma concentration decreased significantly (p < 0.05) from 49 +/- 17 to 35 +/- 16 at 15 days and to 35 +/- 13 nmol/ml at 45 days of therapy (free carnitine) and from 60 +/- 18 to 50 +/- 18 at 15 days and to 55 +/- 14 nmol/ml at 45 days of therapy (total carnitine), whereas acylcarnitine increased significantly (p < 0.05) from 10 +/- 8 to 14 +/- 8 at 15 days and to 18 +/- 16 nmol/ml at 45 days of therapy. Free carnitine urinary excretion decreased significantly (p < 0.05) from 200 +/- 135 to 115 +/- 76 and 118 +/- 75 mumol/24 h, whereas acylcarnitine urinary excretion increased significantly (p < 0.05) from 78 +/- 56 to 154 +/- 98 and 155 +/- 89 mumol/24 h after VPA therapy was started. As a consequence, acylcarnitine renal clearance increased significantly (+30%, p < 0.05) whereas free carnitine renal clearance did not change during VPA therapy. No difference was detected between 15 and 45 days of therapy. No patients experienced symptoms of VPA toxicity. Our results suggest that VPA in patients increases both formation and renal clearance of acylcarnitine. PMID- 8422858 TI - Anticonvulsant action of oxcarbazepine, hydroxycarbamazepine, and carbamazepine against metrazol-induced motor seizures in developing rats. AB - Antimetrazol effects of carbamazepine (CBZ, 5, 12.5, 25, or 50 mg/kg), oxcarbazepine (OCBZ, 5, 10, 30, or 60 mg/kg), and hydroxycarbamazepine (HCBZ, the main human metabolite of OCBZ, 10, 30, or 60 mg/kg) were studied in 7-, 12-, 18-, 25-, and/or 90-day-old laboratory rats. No drug tested affected the incidence of minimal (clonic) metrazol seizures (mMs) in animals aged > or = 18 days; in rats aged 7 or 12 days in which mMs are rare under control conditions, the incidence of mMs was increased by lower doses of CBZ and HCBZ. All drugs tested specifically abolished the tonic phase of major generalized tonic-clonic seizures (MMs) in a dose-dependent manner. In addition, CBZ and OCBZ were able to suppress all phases of MMs in the two youngest groups (7- and 12-day-old). There were no marked differences among the three drugs tested (CBZ, OCBZ, and HCBZ) on their action against metrazol-induced seizures during ontogenesis of rats; i.e., all these drugs appeared to possess an identical profile of anticonvulsant action. PMID- 8422859 TI - Vigabatrin (gamma-vinyl-GABA): neuropathologic evaluation in five patients. AB - We performed neuropathologic examination of cerebral cortex specimens from 4 patients who underwent epilepsy surgery and the brain of 1 patient who died suddenly. All had severe epilepsy and had received gamma-vinyl-GABA (GVG, vigabatrin) for 3-5.5 years. Neither the surgically resected temporal lobe specimens nor the frontal and temporal lobes autopsy specimens showed abnormal white matter vacuolation. PMID- 8422860 TI - Effect of sustained pyridoxine treatment on seizure susceptibility and regional brain amino acid levels in genetically epilepsy-prone BALB/c mice. AB - Epilepsy-prone and epilepsy-resistant substrains were selectively bred from a strain of BALB/c mice; audiogenic-sensitive epilepsy-prone animals showed enhanced sensitivity to chemical convulsants. Treatment with pyridoxine (100 mg/L in drinking water) initiated at mating and continued throughout pregnancy and the life of the offspring abolished the enhanced sensitivity to chemical convulsants and reduced the severity of audiogenic seizures. Withdrawal of pyridoxine restored the enhanced seizure sensitivity. [1H] Nuclear magnetic resonance (NMR) spectroscopy of perchloric acid extracts of tissue was used to determine the concentrations of several compounds [N-acetylaspartate (NAA), GABA, glutamate, aspartate, alanine, taurine, creatine, cholines, inositol] in the hippocampus, neocortex, brainstem, and cerebellum of untreated and pyridoxine-treated 6-week old female animals. The ratios of the concentrations of excitatory to inhibitory putative neurotransmitter amino acids tended to be higher in epilepsy-prone animals, with the most pronounced difference being a significantly elevated glutamate/GABA ratio in every brain region examined. Pyridoxine treatment abolished this imbalance in the hippocampus, brainstem, and cerebellum, but not in the neocortex. Treatment of epilepsy-resistant animals with pyridoxine using the same protocol decreased the glutamate/GABA concentration ratio in the hippocampus, brainstem, and neocortex and resulted in impaired development of the animals. The amino acid imbalance and the accompanying seizure susceptibility in these genetically epilepsy prone mice may originate from an inborn error in pyridoxine metabolism or in a pyridoxine-dependent enzyme system. PMID- 8422861 TI - Rapid cessation of focally induced generalized seizures in rats through microinfusion of lidocaine hydrochloride into the focus. AB - An experimental animal model of complex partial seizures which become secondarily generalized is produced by microinfusion of the GABA antagonist bicuculline (BIC) into the deep prepiriform cortex (DPC) of rats. In the present study, we investigated the effects of microinfusion of the local anesthetic, lidocaine hydrochloride, directly into the BIC focus in the DPC and demonstrated that direct inactivation of the focus arrested a focal seizure that was in progress. A measure of the integrated amplitude of the electrocorticogram (ECoG) and behavioral seizure scores from unanesthetized and freely moving rats were used to address this question quantitatively. Microinfusion of 2% lidocaine hydrochloride into the BIC focus significantly reduced the integrated amplitude of the ECoG to levels that did not differ from baseline in either hemisphere (mean = 112% ipsilateral, 99% contralateral), whereas saline microinfusion had no effect (mean = 175% ipsilateral, 125% contralateral). Moreover, ECoG reductions after lidocaine were present as soon as the microinfusion was complete. Behaviorally, clonic seizure severity was assessed on a rating scale of 0-5. Lidocaine microinfusion significantly reduced the seizure scores to values not different from baseline during the first postinfusion measurement period (i.e., 30 s). Microinfusion of saline alone also significantly reduced behavioral seizure severity, although to a lesser degree and not as rapidly as lidocaine. This effect suggests the need for caution in interpretation and design of studies investigating the anticonvulsant effects of various pharmacologic agents when microinfusions are used. PMID- 8422863 TI - Duration of sodium amytal effect: behavioral and EEG measures. AB - In performance of an intracarotid sodium amytal test duration of action is a critical factor since measurements obtained after cessation of action are invalid. Duration of action is often monitored by measuring handstrength contralateral to injection or by visual inspection of amytal-induced EEG slow waves ipsilateral to injection. We describe new objective methods of monitoring drug effect: quantified EEG and sensory discrimination. In a first study of patients with scalp electrodes, the two traditional and the two new measures were compared in 40 injections. Results indicated that the two EEG measures did not differ significantly and estimated a shorter drug effect than the two behavioral measures, which were also not significantly different. The two new measures had a smaller SD, indicating that they may be less erratic. In a second study of 14 patients, using intracerebral electrodes, we showed that slow waves in the frontocentral region lasted as long as the motor and sensory effects, but also that slow waves in the medial temporal region dissipated earlier than the motor and sensory deficits. Our results suggest that motor and sensory measures may overestimate the time in which valid testing can occur if hippocampal function is at issue. PMID- 8422862 TI - Correlation between function and structure in "epileptic" human hippocampal tissue maintained in vitro. AB - Intracellular and extracellular recordings were obtained from the CA1 region and gyrus dentatus of human hippocampi that had been surgically removed for treatment of intractable seizures. Pathologic diagnoses were obtained, and electrophysiologic results were correlated with degree of principal cell loss (sclerosis) in the regions studied. Extracellular field potentials were absent or smaller in amplitude, with abnormal characteristics, in sclerotic regions. In all regions, regardless of the degree of sclerosis, individual pyramidal or granule cells could be penetrated. These neurons showed apparently normal electrophysiologic characteristics and intact excitatory synaptic input. No signs of hyperexcitability were observed. PMID- 8422864 TI - Hemispheric specialization in children with unilateral epileptic focus, with and without computed tomography-demonstrated lesion. AB - We evaluated 24 testable children with unilateral epileptic foci, with and without radiologically demonstrable lesions, to determine if the normal pattern of cerebral dominance is changed by the presence of an epileptic focus. Verbal and figural stimuli were presented tachistoscopically to the right and left visual hemifields to investigate the specific abilities of the two hemispheres. Three blocks of stimuli consisting of two-letter patterns, three-letter patterns, and meaningful two-syllable words were used as verbal stimuli to assess the abilities of the left hemisphere. A test consisting of localizing a dot on a 3 x 3 matrix was used to assess the abilities of the right hemisphere. Six normal children were chosen as controls. In all groups, specialization of the epileptic hemisphere was lost. The presence or absence of a radiologically demonstrable lesion did not influence this pattern. PMID- 8422865 TI - Value of intraoperative EEG changes during corpus callosotomy in predicting surgical results. AB - The intraoperative transformation of generalized epileptiform discharges (GED) to lateralized epileptiform activity during the course of corpus callosum sectioning for intractable epilepsy in 37 patients was correlated with percentage of decrease in atonic-tonic seizures with "drops" at mean follow-up of 26 months (range 12-86 months). Twenty-seven (73%) patients had intraoperative interictal discharges, and 21 (78%) showed varying degrees of lateralization of GED during corpus callosum sectioning (two thirds to total). All patients experienced > 80% reduction in atonic-tonic seizures with drops. The group (n = 7) with largest decrease in GED had the greatest decrease in seizures (95.5%). Six patients without change in GED had 88% decrease in seizures, as did 14 patients (85-86%) with mild or moderate decreases in GED, but there was no statistically significant correlation between decrease in GED and seizure frequency after operation. Thus, although lateralization of GED after corpus callosum sectioning was evident in 78% of patients with GED, the degree of lateralization of GED did not correlate with degree of reduction of tonic-atonic seizures. Therefore, intraoperative surface EEG monitoring does not appear to be helpful at this time as a guide to extent of callosotomy. PMID- 8422866 TI - Corpus callosotomy for seizures associated with band heterotopia. AB - Band heterotopia is a severe form of neuronal migration disorder associated with intractable epilepsy and neurologic impairment. Surgical treatment of seizures associated with this malformation has not been reported previously. We report a patient with band heterotopia and poorly controlled atonic seizures causing falls and injury. The patient was treated with anterior corpus callosotomy, with significant postoperative decrease in seizure frequency. Corpus callosotomy is a reasonable alternative to consider in management of patients with cortical heterotopia and intractable seizures. PMID- 8422867 TI - Metaphit-induced audiogenic seizure and its inhibition by MK-801: electroencephalographic and behavioral characterization. AB - Adult male Wistar rats were subjected to intense sound stimulation from an electric bell (100 dB and 12 KHz for 60 s) after a single intraperitoneal (i.p. 50 mg/kg) injection of metaphit [1-(1-/3 isothiocyanatophenyl-cyclohexyl) piperidine]. EEG recordings demonstrated appearance of paroxysmal activity and spike-wave complexes from cortical electrodes, with frequency and amplitude increasing with time. Metaphit-induced audiogenic seizures in the rats were tested 24 h after metaphit administration. The seizures consisted of wild running followed by clonic and tonic convulsions, and the seizure pattern could be elicited at hourly intervals for the next 24 h in all tested animals. Forty-eight hours after metaphit administration, susceptibility to sound stimulation began to decrease gradually. The first component of seizure response to disappear was tonic extension, followed by disappearance of clonic convulsion; the last component to disappear was running behavior. Each behavioral seizure response had a characteristic EEG correlate. After approximately 50 h, no animal responded to sound stimulation. The noncompetitive antagonist of the N-methyl-D-aspartate (NMDA) receptors, MK-801 [5-methyl-10,11-dihydro-5H-dibenzo (a,d) cyclohepten 5,10-imine maleate] was evaluated as an anticonvulsant against metaphit-induced audiogenic seizures in two experiments. In the first experiment, MK-801 was administered in a single dose of 0.5 mg/kg i.p. 23.5 h after metaphit injection and 30 min before sound stimulation, which completely blocked both the EEG and the behavioral response to sound stimulation for 37 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8422868 TI - Occipitotemporal epilepsies: evaluation of selected patients requiring depth electrodes studies and rationale for surgical approaches. AB - In 8 patients in whom it was uncertain whether they had occipital or temporal lobe (TL) epilepsy, clinical, scalp EEG, and radiologic features were correlated with the sites of seizure onset as determined by depth EEG. The 8 patients were selected from > 40 with occipital epilepsy because they had (a) an aura considered to be of occipital lobe (OL) origin, (b) an occipital interictal epileptic focus, (c) an OL lesion, or (d) a combination of all of these. Scalp EEG and clinical patterns suggested temporal involvement in all, however. Extracranial EEG recordings were often misleading, showing multilobar interictal epileptic abnormalities, and seizure onset was of poor localizing value and did not clarify the problem sufficiently. Intracranial EEG recordings showed that seizure onset could be ordered along an occipitotemporal gradient. Consistent OL seizure onset was observed in patients who had only elementary visual auras. Those who had inconsistent aura or no aura, suggesting OL origin, had onset of most attacks in the TL. All patients had a seizure spread pattern suggesting early TL involvement. To prevent visual field defect, surgical approaches included temporal resection when temporal seizure origin or spread was demonstrated; although occasionally this produced excellent results, it was of limited benefit in most patients, even when some seizures were proven to originate in TL structures. In patients with malignant epilepsy and in those with an occipital lesion, occipital resection should be considered. PMID- 8422869 TI - Facilitation of infantile spasms by partial seizures. AB - We report 16 patients with infantile spasms in whom onset of the clusters of spasms appeared to be triggered by close temporal association with partial seizures. Common features included the presence of focal cerebral lesions in 12 infants (3 were classifiable as cryptogenic); all had partial seizures with EEG localization, clusters of bilateral spasms always preceded by partial seizures, and response to adrenocorticotropic hormone (ACTH) and traditional antiepileptic drugs (AEDs) generally was poor. Three had complete agenesis of the corpus callosum, which argues against interhemispheric callosal spread of focal discharges resulting in the generalized spasms. Surgical cortical resections were performed in 6 of the infants, with good outcomes. This group of patients supports a model in which the spasms, although probably generated at a subcortical level, are facilitated or possibly induced by focal discharges from cortical pathology. PMID- 8422870 TI - Iodine deficiency disorders and endemic goitre. PMID- 8422871 TI - Dietary determinants of serum beta-carotene and serum retinol. AB - The relationship of major dietary carotenoids, preformed and total vitamin A, and different foods to serum beta-carotene and serum retinol levels was studied among 224 male and 117 female adults taken from the Finnish Mobile Clinic Health Examination Survey. Serum nutrients were analysed after 10-15 years of storage at -20 degrees C. Dietary data were collected by a quantitative dietary history interview method. Intakes of nutrients were calculated based on analysed data on Finnish foods. The positive gradient between beta-carotene intake and serum level, being highly significant in women and non-significant in men, was concentrated in non-smokers. Other major dietary carotenoids tended to be positively correlated with serum beta-carotene in parallel with dietary beta carotene. Carrot intake was the most specific single food predictor for serum beta-carotene. Serum retinol levels were not significantly associated with dietary variables and were not affected by current smoking. In women, serum beta carotene values were higher, but serum retinol levels lower, compared with men. The results support earlier findings that smoking modifies the association between dietary beta-carotene and serum beta-carotene, and suggest that despite the long storage of serum samples beta-carotene determinations had some value as a biological marker for beta-carotene in the diet. PMID- 8422873 TI - Alcohol consumption, smoking habits and body fat distribution in Italian men and women aged 20-60 years. AB - The study was performed on 601 patients (294 males and 307 females) of a general practitioner. Alcohol intake and smoking habits were compared with other anthropometric measurements including waist-to-hip girth ratio. Patients were divided into non-smokers and smokers (subdivided into three groups according to the number of cigarettes smoked per day) and into non-drinkers and drinkers (subdivided into three groups with different alcohol intakes). Ex-smokers were excluded from the study. Analysis of covariance using age, body mass index, physical activity and menopausal status as covariates, showed that: (1) cigarette smoking is not accompanied by a specific pattern for body fat distribution; (2) waist-to-hip ratio was significantly different for the four classes of alcohol intake for women (non-drinkers: 0.809, < 11 g: 0.805, 11-20 g: 0.809, > 20 g: 0.826; F = 2.8, P < 0.05) but not for men (non-drinkers: 0.944, < 20 g: 0.934, 21 40 g: 0.940, > 40 g: 0.943; F = 0.9); (3) increased alcohol intake corresponds to an increased lipid and energy supply. PMID- 8422872 TI - Growth dynamics during the first two years of life: a prospective study in the Philippines. AB - This study examines determinants of growth from birth to 24 months in a sample of approximately 3000 urban and rural Filipino children. Individual, household, and community data were collected bimonthly during the Cebu Longitudinal Health and Nutrition Survey. Separate longitudinal, multivariate models were used to identify determinants of weight in children from birth to 6 months and 6-24 months of age. Previous weight, male gender, mother's height, and season of the year showed significant positive associations with weight in all models. Full and mixed breast-feeding significantly increased weight, but the effects of breast feeding declined as children got older. Breast-feeding had a direct growth enhancing effect in addition to its indirect effect through the prevention of diarrheal morbidity. Detrimental effects of recent diarrheal morbidity were particularly important in the older age group, but these effects were mitigated by breast-feeding. Since infant feeding variables are included in the models, the results strongly suggest an effect of diarrheal morbidity on growth independent of its known effects on infant feeding and dietary intake. Febrile respiratory infections had important detrimental effects on weight in both age groups. PMID- 8422874 TI - Bioimpedance analysis and resting energy expenditure in undernourished and refed anorectic patients. AB - Anthropometry, bioimpedance analysis (BIA) and resting energy expenditure (REE) were evaluated in young female patients affected by anorexia nervosa who were either severely malnourished (MnA) or already refed (RfA) and in a control group of healthy young women (WnC). Fat-free mass (FFM) and fat mass (FM), evaluated from skinfold thickness, were severely decreased in the MnA group while they were very similar in the RfA group and in the WnC group. With respect to BIA parameters, impedance (Z) was significantly higher in absolute terms in the MnA but lower than in the other groups when expressed as specific impedance, i.e. after normalization for both FFM and height. Phase angle significantly differed between the three groups, being significantly (P < 0.01) lower in the MnA (3.70 +/- 0.83 degrees) and the RfA (4.36 +/- 0.82) than in the WnC (5.17 +/- 0.40). REE was comparable in RfA subjects and WnC subjects, while it was sharply decreased (P < 0.01) in the MnA patients both in absolute value and after adjustment for body composition (FFM and FM) or body weight. This cross-sectional study shows that marked changes in BIA parameters occur in undernourished anorectic patients and also in the anorectic subjects who were previously very underweight but studied only after having already regained a normal body size. REE was deeply decreased in the undernourished anorectic women even when the differences in body composition or Wt were taken into account, indicating the occurrence of a significant adaptation of energy expenditure to chronic underfeeding. PMID- 8422875 TI - Can infants and young children eat enough green leafy vegetables from a single traditional meal to meet their daily vitamin A requirements? AB - To evaluate the feasibility of providing adequate vitamin A precursors to meet the daily need from a meal oftraditionally cooked green leafy vegetables and boiled rice and to understand mothers' perceptions and acceptance of leafy vegetables for infants and young children, 118 children aged 6 months to 3 years and their mothers were studied. The mothers were interviewed regarding their acceptance and perceptions about giving leafy vegetables to their young children. Their children were served a measured amount of rice and cooked leafy vegetables and mothers were asked to feed the child within about half an hour. Median intakes of leafy vegetables in children aged 6-11 months, 12-17 months and 18-35 months were 41 g, 71 g and 129 g respectively (in terms of raw green leaf). Approximately 40 g green leaf provides the recommended daily allowance for vitamin A precursors. 77% of the under-1-year-old children were breast-fed. The breast-fed children had a lower intake of vegetables than the completely weaned children. 87% of the children were found to like vegetables, 89% of the mothers liked to give vegetables to their children and 74% of the mothers answered that vegetables were good for health. Only two mothers (1.5%) refused to feed their children the leafy vegetables. The results show that leafy vegetables are acceptable to most of the mothers, and that children can eat enough leafy vegetables to meet a day's need of vitamin A precursors. Feasibility of feeding children enough green leafy vegetables at home on a regular basis needs further study. PMID- 8422876 TI - Body mass index, ageing and differential reported morbidity in rural Sarawak. AB - Classifications of adult low energy nutritional status based on the Quetelet or body mass index (weight/height2) have often assumed independence of age and sex. This paper reports findings of a study of 447 men and 564 women aged over 18 years and belonging to the Iban tribe of central Sarawak, East Malaysia. BMI and fat-free mass fell markedly in both sexes, and fat mass in women but not men, after about 40 years of age. In men over age 40, and women aged 18-40, BMI was sensitive to reported morbidity. For subjects aged over 40 years, BMI was related to morbidity independently of age effects in men, and to age alone in women. These findings suggest that the functional significance of low BMI differs between the sexes and with age. PMID- 8422877 TI - Exercise-associated myopathy: is calcium the culprit? PMID- 8422878 TI - Muscle histopathology and plasma aspartate aminotransferase, creatine kinase and myoglobin changes with exercise in horses with recurrent exertional rhabdomyolysis. AB - Six horses with a history of recurrent exertional rhabdomyolysis (RER) (Horses A F) and 7 control horses performed a submaximal and later a near-maximal treadmill exercise test. Blood samples were obtained before, during and after exercise and muscle biopsies were taken before and after exercise. At rest, plasma aspartate aminotransferase (AST) activities in horses with RER were above 95% confidence intervals for control horses. During submaximal exercise, 3 horses with RER (A, B and C) had much greater increases in plasma AST, creatine kinase (CK) and myoglobin concentrations than did Horses D, E and F and control horses. Clinical signs of muscle stiffness and pain were only obvious in Horse A. During near maximal exercise, only Horse C showed a substantial increase in CK activity and myoglobin concentrations without any associated clinical signs of rhabdomyolysis. Muscle biopsies from Horses A, B and C contained necrotic type II fibres which, on electron microscopic examination, contained disrupted myofibrils and swollen mitochondria. These results suggest that, in RER, subclinical episodes of muscle fibre necrosis and associated increases in plasma AST, CK and myoglobin occur with exercise more frequently than could be detected clinically. Furthermore, the pattern of increase in muscle enzymes and myoglobin concentrations in the 6 horses with RER suggested that the high plasma AST and CK activities commonly observed at rest in symptom-free Standardbred horses are probably a result of repeated subclinical episodes of rhabdomyolysis after exercise, rather than leakage due to abnormal sarcolemmal permeability. PMID- 8422879 TI - Blood chemistry and skeletal muscle metabolic responses to exercise in horses with recurrent exertional rhabdomyolysis. AB - Six horses with a history of recurrent exertional rhabdomyolysis (RER) and 7 control horses performed both a 55-min submaximal and later a 10-min near-maximal exercise test on a treadmill. Blood samples were obtained during exercise and gluteus medius muscle biopsies were obtained before and immediately after each exercise test and at 24 h after completion of the submaximal test. Rhabdomyolysis was developed by 3 of 6 RER horses during submaximal exercise and in 1 of the RER horses during near-maximal exercise. Concentrations of potassium, glucose, free fatty acids, ammonia, lactate, cortisol, adrenaline and noradrenaline in the blood were measured. None of these variables appeared useful in predicting which RER horses would develop rhabdomyolysis. The RER horses that developed rhabdomyolysis (RERa) had higher cortisol and blood glucose concentrations but otherwise had blood chemistry and muscle metabolic responses during submaximal and near-maximal exercise similar to those of RER horses which did not develop rhabdomyolysis (RERb) and to controls. At rest, muscle glycogen concentrations were significantly higher (> 650 mmol/kg dry wt) in RERa and RERb horses than in controls. Lactate concentrations in muscle after submaximal and near-maximal exercise were similar or lower, respectively, in RERa horses compared with controls. The results of this study indicate that, although horses with RER had high resting intramuscular glycogen concentrations, rhabdomyolysis did not appear to be caused by an excessively rapid rate of anaerobic glycolysis with lactate accumulation. PMID- 8422880 TI - Ultrasonographic and histopathological findings in equine superficial digital flexor tendon injury. AB - The ultrasonographic and histopathological findings in 12 normal and 28 injured superficial digital flexor tendons, with lesions ranging in duration from 2 days to 15 months, were compared. A consistent relationship between the ultrasonographic and histological findings was demonstrated. The echogenicity of lesions, the distinctness of their delineation from the surrounding tissue, and the presence and arrangement of the linear echoes were useful features by which to assess the ultrasonograms. Acute lesions were anechoic, a complex mixture of anechoic and hypoechoic areas, or diffusely hypoechoic. These appearances represented haemorrhage, fibrolysis and early granulation tissue. Fibroplasia and granulation tissue produced well to moderately well defined hypoechoic lesions. Chronic fibrosis was characterised by heterogeneously echogenic areas which were poorly defined from the surrounding tissue and had irregularly-arranged linear echoes on longitudinal images. Intratendinous scar formation resulted in multiple hyperechoic foci. Extensive peritendinous lesions were readily apparent on ultrasonograms, but intertendinous adhesions were more difficult to assess, and produced ill-definition of the borders between the superficial and deep digital flexor tendons. PMID- 8422881 TI - An investigation of injection techniques for local analgesia of the equine distal tarsus and proximal metatarsus. AB - A positive radiographic contrast agent was injected into the tarsometatarsal (TMT) joints of both hindlimbs of 10 horses. Lateromedial radiographic views were obtained at 5, 15 and 30 mins after injection. Injection was successful in 19 of 20 limbs. Communication between the centrodistal (CD) and TMT joints was identified in 7 limbs (35%). Contrast agent extended around the tendons of tibialis cranialis (TC) and fibularis tertius (FT) in 18 limbs, and in 7 limbs some contrast entered the tarsal sheath. Slight to moderate plantar and/or distal extension of contrast agent was identified in 13 limbs. On a subsequent occasion positive contrast agent was injected subtarsally using one of two techniques and radiography was repeated. Contrast agent was principally distributed on the plantar aspect of the 3rd metatarsal bone, the plantar aspect of the suspensory ligament and between the superficial and deep digital flexor tendons. Extension of contrast agent into the TMT joint was identified in only 1 limb but in 8 limbs contrast agent extended into the tarsal sheath. The practical implications of these results include the possibility that local anaesthetic solution injected into the TMT joint may alleviate pain from the CD joint, the insertions of TC and FT or the tarsal sheath. It may also result in perineural analgesia of the dorsal metatarsal nerves or the plantar metatarsal nerves. In some cases subtarsal injection of local anaesthetic solution may result in alleviation of pain from the tarsal sheath. False negative results for subtarsal analgesia may be achieved by inadvertent injection into the tarsal sheath or into a blood or lymphatic vessel. PMID- 8422882 TI - Arterio-venous differences of NEFA during exercise. PMID- 8422884 TI - A method for loading equine platelets with the fluorescent calcium indicator Fura 2: ADP induces a rise in the cytosolic free calcium ion concentration. AB - Equine platelets in platelet-rich plasma were incubated with the fluorescent indicator dye, Fura-2-AM (Fura-2-acetoxymethyl ester) and the degree of loading of the cells with the dye and the extent of hydrolysis of the ester was assessed by quantitative fluorimetry and by thin-layer chromatography respectively. Under these conditions the cells loaded poorly with Fura-2 to a concentration of 4 microM. The technique was validated by demonstrating adequate loading of human platelets with Fura-2, to a concentration of 250-300 microM, using the same method. The removal of plasma from the extracellular medium was important for successful loading of the cells with the dye since washed cells resuspended in a physiological salt solution loaded adequately with Fura-2 to a concentration of 190 microM. Cells loaded as such showed a resting [Ca2+]i of 129 nM and a concentration-dependent rise in [Ca2+]i to a transient maximum of 350 nM when stimulated by the pro-aggregatory agonist adenosine 5'-diphosphate (ADP). PMID- 8422883 TI - Nutrient utilisation by the hindlimb of thoroughbred horses at rest. AB - Nutrient uptake by the hindlimb was investigated utilising the arteriovenous difference technique in 5 Thoroughbred horses fed to maintenance a diet of 100% roughage or 52% oat grain and 48% roughage. Arterial blood was obtained from a catheter inserted into the carotid artery while venous blood was simultaneously collected from a catheter placed into the iliac vein via the medial saphenous vein. The arteriovenous difference for glucose was significant and represented a mean extraction of 10 +/- 1% with no effect of diet. If fully oxidised, glucose uptake (corrected for lactate and pyruvate arteriovenous difference) was sufficient to account for 78 +/- 13% or 107 +/- 15% of the oxygen consumed by the hindlimb in horses fed a roughage or 52% oat grain diet respectively. Acetate was also a major metabolite of the hindlimb, showing a 39 +/- 5% extraction with no effect of diet. However, the 52% oat grain diet did induce a significant decline in the concentration of acetate in arterial blood. The potential contribution to oxidation in the hindlimb was significantly reduced from 32 +/- 4% in horses fed roughage to 21 +/- 3% when fed 52% oat grain. D-3-Hydroxybutyrate uptake could account for 9 +/- 1% of the oxidation by the hindlimb with no effect of diet. The technique for measuring nutrient uptake across the hindlimb using the arteriovenous difference is relatively simple and would be valuable in investigating fuel use by muscle during exercise. PMID- 8422885 TI - Effect of carnitine supplement to the dam on plasma carnitine concentration in the sucking foal. AB - The changes in carnitine in plasma and milk during the first 3 months of lactation were studied in 14 broodmares and their foals. Six of the mares (Group S) were given a supplement of 10 g carnitine split between the morning and evening feeds, starting 2 weeks before birth. At birth the plasma carnitine concentration in Group S mares was about twice that in Group NS mares (no supplement). In both groups the concentration initially declined in the days after birth. Whilst this trend was reversed in Group S mares, the concentration in Group NS mares remained at a reduced level for the remainder of the study. Milk concentrations declined continuously over the monitoring period in both groups. There was no apparent relationship between milk and plasma concentrations. Despite this the milk concentration tended to be higher in Group S than in Group NS mares although differences were not significant. There was an immediate drop in the plasma concentration in foals after birth which was reversed in foals of Group S mares but not in those of Group NS mares. There were no apparent side effects of carnitine supplementation. PMID- 8422886 TI - The correlation of running ability and physiological variables in thoroughbred racehorses. AB - The running abilities of 25 Thoroughbred racehorses were measured at distances of 1200, 1600 and 20000 m. Various physiological variables were measured subsequently on the treadmill and correlated with running speed. There was a negative correlation for running speed with the velocity (VLa4) and work rate (WLa4) at which blood lactate reaches a steady-state concentration of 4 mmol/litre and a positive correlation with peak plasma lactate, suggesting that plasma lactate concentrations of faster horses rise more rapidly and to higher levels than do those of slower horses. The correlation between running speeds and heart rates (HR) was stronger for unfit than fit horses, suggesting that cardiovascular effects of training are more beneficial to slower horses. The significant correlation between running speeds and V200 suggests that the HR of faster horses increases more rapidly than in slower horses performing similar exercise. The correlation of running speeds and VO2max suggests that the HR of faster horses increases more rapidly than in slower horses performing similar exercise. The correlation of running speeds and VO2max suggests that faster horses utilise more oxygen during maximal intensity exercise. The correlation of running speeds with minimum pH and minimum HCO3- suggests that faster horses maintain speed at higher hydrogen ion (H+) concentrations. Correlations between running speeds and the measured variables were consistently stronger for the longer distance runs. Because VLa4 and WLa4 were obtained during sub-maximal exercise, these variables were determined to be the best correlates of running performance. PMID- 8422887 TI - Assessment of calcium dynamics in platelets. PMID- 8422888 TI - A retrospective evaluation of laminitis in horses. AB - Case records of 202 horses treated for laminitis were reviewed with the intent of determining the long-term outcome and correlating this with digital radiographic findings and with the degree of pain associated with the laminitis. At long-term follow-up 57 horses had returned to athletic soundness (Group 1), 20 horses were intermittently lame (Group 2), 19 horses had permanent severe lameness (Group 3), 97 were dead (Group 4), and 9 were lost to follow up. Using simple regression analysis, functional outcome did not correlate with the degree of rotation (R2 = 0.004) or the presence of distal displacement (R2 = 0.139). Functional outcome did correlate with the clinical grade of laminitis (R2 = 0.504). Horses in Group 1 had significantly less distal phalangeal rotation (5.89 +/- 6.48 degrees) than did horses in Group 2 (11.10 +/- 8.19) and Group 3 (14.50 +/- 10.80), but were not significantly different from Group 4 horses (7.49 +/- 6.57). Of 96 surviving horses, 23 had evidence of distal displacement compared with 54 of 97 non survivors. Based on these results, horses that develop distal displacement of the distal phalanx are more likely to die than are horses without distal displacement; however, the presence or absence of distal phalangeal displacement and the degree of distal phalangeal rotation cannot be used to predict the outcome of a horse with laminitis. Clinical assessment is a more reliable means of determining the final outcome and should be given precedence over radiographic findings. PMID- 8422889 TI - Advancing the time of ovulation in the mare with a short-term implant releasing the GnRH analogue deslorelin. AB - A small, biocompatible and short-term implant releasing 1.5 mg or 2.25 mg of the GnRH analogue deslorelin was evaluated in 140 Hanoverian (warm blooded) mares during the 1990 breeding season (Study I). Mares in oestrus and with a follicle 40 +/- 2 mm in diameter were assigned alternately to treatment (70) or remained as untreated controls. Implants were administered subcutaneously, and intervals to ovulation determined by rectal examination and ultrasound at 12-h intervals. Since results with both doses of deslorelin were similar, data were pooled. Deslorelin implantation resulted in ovulations in 65 of 70 mares within 48 h (93%), while only 5 of 70 control mares ovulated within the same time period (7%) (P < 0.01). Most induced ovulations (63%) occurred 36-48 h after implantation. In Study II, 4 groups of 12 Hanoverian mares each were treated with 3,000 or 5,000 iu hCG, or a 2.25 mg deslorelin implant, or received placebo. All treatments resulted in 100% ovulations within 48 h, versus 25% ovulations in controls (P < 0.01), and 63%, 75% and 86% of these ovulations occurred 36-48 h after treatment with 3,000 and 5,000 iu hCG and deslorelin respectively. Hormonal response to deslorelin in treated mares, sampled every 12 h, consisted of elevations of FSH and LH concentrations of > 200% and > 300% baseline values at 12 h (P < 0.001), of 67% and 79% at 24 h (P < 0.01) and of 35% and 49% at 48 h (P < 0.05), respectively. No local reactions at the implantation site were observed. PMID- 8422890 TI - Muscle fibre compartmentalisation in the gluteus medius of the horse. PMID- 8422891 TI - Serum concentrations of ormetoprim/sulphadimethoxine in 1-3-day-old foals after a single dose of oral paste combination. PMID- 8422892 TI - Fracture of the caudal medial femoral condyle in a horse. PMID- 8422893 TI - Scleral mastocytosis in a horse. PMID- 8422894 TI - Did vaccinia virus come from a horse? PMID- 8422895 TI - Cholestatic hepatopathy, thrombocytopenia and lymphopenia associated with iron toxicity in a thoroughbred gelding. PMID- 8422896 TI - Unlike morphine the endogenous enkephalins protected by RB101 are unable to establish a conditioned place preference in mice. AB - The mixed inhibitor prodrug, RB101, was used to study the motivational properties of the endogenous opioid peptides, the enkephalins. In the conditioned place preference test, which is commonly used to investigate the reinforcing properties of drugs, mice alternately treated with morphine (3 mg/kg i.p.) on the initially non-preferred compartment and with saline on the preferred one, for four place pairings, spent more time in the drug-associated compartment. This shift in place preference after the conditioning procedure was not found after treatment with RB101 (80 mg/kg i.p.). Administration of naloxone (1 mg/kg s.c.) after the conditioning phase increased the preference for the drug-associated compartment of mice treated with 6 mg/kg (i.p.) of morphine. This illustrates the negative motivational properties of morphine withdrawal or the establishment of psychic dependence on the drug. In contrast, no modification of preference was observed after injection of naloxone in animals treated with a high dose of RB101 (160 mg/kg i.p.). The failure to establish conditioned place preference by inhibiting endogenous enkephalin metabolism, and the lack of development of psychic dependence after RB101 administration demonstrate for the first time the interest of mixed inhibitors of enkephalin-degrading enzymes as potent new non-addictive analgesics. PMID- 8422897 TI - L-arginine potentiates excitatory amino acid-induced seizures elicited in the deep prepiriform cortex. AB - Microinjection of N-methyl-D-aspartate (NMDA; 1 and 2.5 nmol) or kainate (KA; 50 pmol) into the deep prepiriform cortex elicited behavioral signs of seizure activity. No epileptiform activity was observed after deep prepiriform cortex microinjection of either L-arginine (L-Arg, 5 and 10 nmol) or its D-enantiomer, D arginine (D-Arg, 2.5-10 nmol). However, both the seizure score and the incidence of electroencephalographic (EEG) epileptic discharges elicited by NMDA (1 and 2.5 nmol) and KA (50 pmol) were significantly increased by L- but not D-Arg. The facilitatory effects of L-Arg on seizure activity elicited by both NMDA and KA were dose-dependent and could be prevented by co-administration of L-Arg (10 nmol) and the nitric oxide (NO) synthase inhibitor, N omega-nitro-L-arginine methyl ester (L-NAME, 20 nmol). Motor and electrocortical seizures were observed after microinjection of the NO donor sodium nitroprusside (SNP; 5 to 20 nmol) into the deep prepiriform cortex. Infusion of methylene blue (20 nmol), a soluble guanylate cyclase inhibitor, protected against SNP-induced seizures. Furthermore, prior infusion of a subconvulsant dose of SNP into the deep prepiriform cortex significantly potentiated the seizure activity elicited by either NMDA (1 and 2.5 nmol) or KA (50 pmol). These results support the proposal that NO is formed from L-Arg upon excitatory amino acid receptor activation within the deep prepiriform cortex, thereby contributing to the genesis of seizure activity. PMID- 8422898 TI - Neurotensin and cholecystokinin octapeptide control synergistically dopamine release and dopamine D2 receptor affinity in rat neostriatum. AB - Combined perfusion of the neostriatum with 1 nM of cholecystokinin octapeptide (CCK-8) and 0.01, 0.1 or 1 nM of neurotensin was done in the halothane anesthetized rat after systemic apomorphine treatment (0.05 mg/kg, s.c.). Neurotensin (1 nM) plus CCK-8 (1 nM) effectively counteracted the apomorphine induced inhibition of neostriatal perfusate levels of dopamine (DA). With a constant concentration of CCK-8 (1 nM), the apomorphine-induced inhibition of DA release was counteracted dose relatedly by neurotensin in concentrations of 0.01, 0.1 and 1 nM. The results of binding experiments demonstrated that threshold concentrations of CCK-8 and neurotensin significantly increased the KD values of the high-affinity D2 receptors without significant alterations in the low affinity D2 receptors or in the proportion of D2 receptors in the high-affinity state. Thus, neurotensin and CCK receptors may regulate synergistically, via intramembrane interactions with the D2 receptors, the binding characteristics and the signal transduction of D2 autoreceptors in the neostriatum. The combined presence of very low concentrations of CCK-8 and neurotensin in the extracellular fluid may be sufficient to regulate D2 receptor transduction, underlining the important role of these peptide receptor interactions with the D2 receptors. PMID- 8422899 TI - The influence of pH on the use-dependent effects of lidocaine in adult and neonatal canine Purkinje fibers. AB - We used microelectrode techniques to examine the influence of pH on the effects of lidocaine on neonatal and adult canine Purkinje fiber action potentials. Lidocaine, 5 mg/l and 10 mg/l, significantly decreased overshoot, Vmax, and action potential duration in neonatal and adult fibers (basic cycle length 1300 and 300 ms) at pH 7.3 and 6.8. The effects were similar except for that on action potential duration which was greater in adults. Lidocaine, 5 mg/l, caused a comparable tonic block in adults and neonates at pH 7.3 (12 +/- 2 and 11 +/- 2%, respective decreases of Vmax) and at pH 6.8. Onset of use-dependent block (UDB) (pH 7.3) was faster in adults than neonates, 2 +/- 0.3 vs. 6 +/- 0.8 beats (P < 0.05); and tau off was slower in adults (133 +/- 10 ms) than neonates (81 +/- 8 ms; P < 0.05). At pH 6.8 the 'on' rates were 5 +/- 0.8 and 7 +/- 1 beats for adults and neonates, respectively (P > 0.05), and tau off increased to 210 +/- 15 ms for adults and 193 +/- 10 ms for neonates (P > 0.05). Thus, developmental differences in lidocaine action may be modified by the degree of protonation. PMID- 8422900 TI - Age-related effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine treatment of common marmosets. AB - The effect of treatment with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on juvenile (6-8 months), young adult (2-4 years) and aged (8-10 years) common marmosets were compared. Juvenile marmosets were more resistant to the actions of MPTP and required a greater cumulative dose over a longer period to induce the same degree of motor disability observed in older animals. Young adult and aged marmosets showed an equivalent motor recovery in the 4-5 weeks following cessation of MPTP treatment, but juvenile animals were less able to compensate for the motor impairments. Losses of putamen [3H]dopamine uptake and caudate nucleus dopamine content were equivalent in young adult and aged animals. However, juvenile animals showed a more marked degree of dopamine depletion and reduction in [3H]dopamine uptake. Histological analysis showed cell loss in the substantia nigra to be most prominent in juvenile animals although it was evident in all groups. No loss of cells in the locus coeruleus was apparent in any of the groups studied, and no intraneuronal eosinophilic inclusions were seen. Greater nigral cell loss and dopamine depletion were required in juvenile animals to impair motor function. The degree of behavioural recovery was less in juvenile animals than in young adult and aged marmosets. The extent of behavioural recovery appeared linked to the severity of cell loss and was not reduced in old age. PMID- 8422901 TI - In vivo occupancy of the striatal dopamine uptake complex by various inhibitors does not predict their effects on locomotion. AB - We compared the ability of various dopamine (DA) uptake inhibitors to displace the in vivo striatal [3H]GBR 12783 (1-[2(diphenylmethoxy) ethyl)-4-(3-phenyl 1[3H]-2-propenyl)-piperazine) binding with their stimulant effect on locomotor activity on mice. GBR 12783 (8 mg/kg), GBR 13069 (10 mg/kg), cocaine (20 mg/kg), mazindol (3 mg/kg) or pyrovalerone (2 mg/kg) stimulated locomotion as long as they occupied the DA uptake complex. In contrast, nomifensine (3 mg/kg) did not stimulate locomotion although it competed with [3H]GBR 12783 for the occupancy of the DA uptake complex at a significant level (> 50%). Administered at their ED50 doses, GBR 12783, BTCP (N-[1-(2-benzo(b)thiophenyl)cyclohexyl]piperidine, GBR 13069, amineptine and dexamphetamine significantly increased locomotor activity whereas the other inhibitors tested did not. The locomotor response elicited by GBR 12783 (10 mg/kg) was not decreased by desipramine (20 mg/kg) nor by oxaprotiline (10 mg/kg). The increase in locomotion elicited by GBR 12783 was positively correlated with the basal locomotor activity of the mice. The stimulant effect of GBR 12783 was potentiated by SKF 525A and by budipine. Additional pharmacological properties might conceal the relationship between the effects of some DA uptake inhibitors on locomotion, and on in vivo occupancy of DA uptake sites. PMID- 8422902 TI - ATP-dependent K+ channel blockers antagonize morphine- but not U-504,88H-induced antinociception. AB - The effects of four ATP-dependent K+ channel blockers (hypoglycemic sulfonylureas) against morphine- and U50488H-induced antinociception were evaluated using the tail flick test in mice. None of the sulfonylureas tested significantly modified tail flick latency in control animals. However, i.c.v. pretreatment with gliquidone (0.4-1.6 micrograms/mouse), glipizide (2.5-10 micrograms/mouse), glibenclamide (10-40 micrograms/mouse) or tolbutamide (20-80 micrograms/mouse) dose dependently antagonized morphine-induced antinociception approximately equieffectively, the only difference being in potency: gliquidone > glipizide > glibenclamide > tolbutamide. This effect of sulfonylureas was very specific, since none antagonized the antinociception elicited by U50488H even at doses twice as great as the dose that induced maximum antagonism of morphine antinociception. Because morphine, but not U50488H, opens K+ channels in neurons and because the order of potency of the different sulfonylureas for blocking ATP dependent K+ channels in neurons and for antagonizing morphine antinociception is the same, we suggest that morphine antinociception is mediated by the opening of ATP-dependent K+ channels. PMID- 8422903 TI - Effect of riluzole on focal cerebral ischemia in rats. AB - The effects of riluzole, a putative inhibitor of glutamate release, on the histological and neurobehavioral consequences of middle cerebral artery occlusion were tested in Sprague-Dawley rats. Neurobehavioral studies (neurological examination, passive avoidance task) were carried out with sham-operated and occluded rats. Riluzole 4 and 8 mg/kg administered 30 min after occlusion reduced (P < 0.01) the cortical infarct (respectively 94 +/- 12 mm3 and 73 +/- 15 mm3 versus 139 +/- 8 mm3 for control rats). Striatum necrosis was not modified by the low dosage (46 +/- 3 mm3 versus 49 +/- 3 mm3 for control rats), whereas the high dosage increased it (61 +/- 3 mm3, P < 0.05). The ischemia-induced neurological and memory impairments were not improved by riluzole. Our results indicate that a drug depressing glutamatergic neurotransmission without blocking the glutamate receptors exerts anti-ischemic activity. Moreover, the results highlight the need for carrying out histological and neurobehavioral studies in parallel in this model. PMID- 8422904 TI - Activation of K+ channel in vascular smooth muscles by cytochrome P450 metabolites of arachidonic acid. AB - Arachidonic acid can be oxidatively metabolized by cytochrome P450 epoxygenase to four regioisomeric epoxyeicosatrienoic acids (5,6-; 8,9-; 11,12-; 14,15-EET), which exhibit vasorelaxant effects in vivo and in vitro with unknown mechanisms. In this study, the patch-clamp method was used to examine the effects of EETs on the Ca(2+)-activated K+ channel in cells from rabbit portal vein, rat caudal artery, guinea pig aorta and porcine coronary artery. In all four cell types, EETs in the bath activated the K+ channel in cell-attached patches by increasing the single channel open-state probability. Potencies of the four EETs did not differ significantly for each cell type. The concentrations for doubling open state probability were 0.1 microM in portal vein and coronary artery, 0.3-1 microM in aorta and 1-3 microM in caudal artery. In caudal artery cells, K+ channel activation by 3 microM 5,6- and 1 microM 11,12-EET was blocked and reversed by glyburide at 0.5 microM. In aorta, coronary artery, and caudal artery cells, micromolar EETs induced a dose-dependent and reversible augmentation of whole-cell K+ current by 50-120% and a 5-12 mV hyperpolarization. EETs on the cytosolic side of inside-out patches produced little or no potentiation of K+ channels, implying an interaction of receptor-mediated nature. Thus, EETs may promote vasodilation by functioning as endogenous K+ channel openers. PMID- 8422906 TI - Repeated methamphetamine-treatment alters brain sigma receptors. AB - The present study was undertaken to examine whether repeated exposure of rats to the psychostimulant drug, methamphetamine, induces changes in sigma receptor binding. A significant up-regulation (130-145% of control Bmax) of sigma receptors, labeled with [3H](+)pentazocine, was observed in the substantia nigra, frontal cortex and cerebellum of rats treated with methamphetamine (4.0 mg/kg per day; 10 days). These findings suggest that methamphetamine-induced psychosis may be associated with the up-regulation of sigma receptors in critical brain regions. PMID- 8422905 TI - Region-specific inhibition of potassium-evoked [3H]noradrenaline release from rat brain synaptosomes by neuropeptide Y-(13-36). Involvement of NPY receptors of the Y2 type. AB - The effects of the Y2 receptor agonist neuropeptide Y NPY-(13-36) on the depolarization-evoked release of [3H]noradrenaline (NA) from synaptosomal preparations of the medulla oblongata, the hypothalamus, the hippocampal formation and the parieto-occipital cortex of the male rat were studied. NPY-(13 36) (0.1-100 nM) caused a concentration-related inhibition of the depolarization induced release of [3H]NA in all areas studied, except the parieto-occipital cortex. The results indicate that NPY Y2 receptors are present on NA terminals in all areas studied, except the parieto-occipital cortex and inhibit depolarization evoked [3H]NA release. PMID- 8422907 TI - Formation of a novel 20-hydroxylated metabolite of lipoxin A4 by human neutrophil microsomes. AB - Lipoxin A4 (LXA4) is a biologically active compound produced from arachidonic acid via interactions of lipoxygenases. Incubation of LXA4 either with human neutrophils or with the neutrophil microsomes leads to formation of a polar compound on a reverse-phase high-performance liquid chromatography. We have identified the metabolite as 20-hydroxy-LXA4, a novel metabolite of arachidonic acid, on the basis of ultraviolet spectrometry and gas chromatography-mass spectrometry. The LXA4 omega-hydroxylation requires both molecular oxygen and NADPH, and is inhibited by carbon monoxide, by antibodies raised against NADPH cytochrome P-450 reductase, or competitively by leukotriene B4 (LTB4) and LTB5, substrates of LTB4 omega-hydroxylase. These findings indicate that the formation of 20-hydroxy-LXA4 is catalyzed by a neutrophil cytochrome P-450, the LTB4 omega hydroxylase. PMID- 8422908 TI - Expression and targeting of a 47 kDa integral peroxisomal membrane protein of Candida boidinii in wild type and a peroxisome-deficient mutant of Hansenula polymorpha. AB - A 47 kDa integral peroxisomal membrane protein (PMP47) of Candida boidinii was expressed in wild type (WT) and a temperature-sensitive (Ts6) peroxisome deficient (per) mutant of Hansenula polymorpha. The subcellular location of PMP47 appeared to be dependent on the level of expression. At low expression levels PMP47 was sorted to the peroxisomal membrane; however, in Ts6 cells grown at restrictive temperatures (which lack intact peroxisomes) PMP47 was solely located in small cytosolic aggregates, together with homologous H. polymorpha PMP's. At enhanced expression levels, however, part of the protein also became incorporated into mitochondria, both in transformed WT and Ts6 cells. PMID- 8422909 TI - Membrane topology of the 22 kDa integral peroxisomal membrane protein. AB - In order to study the membrane topology and the possible function of the rat liver 22 kDa integral peroxisomal membrane protein (PMP 22) at a molecular level, we have cloned PMP 22 from a lambda gt11 expression library and sequenced its cDNA. Hydropathy analysis of the deduced primary structure indicates 4 putative transmembrane segments. The accessibility to exogenous aminopeptidase of PMP 22 in intact peroxisomes suggests that the N-terminus faces the cytosol. A model of the topology of PMP 22 in the peroxisomal membrane is discussed. Homology studies revealed a striking similarity with the Mpv 17 gene product. Lack of this membrane protein causes nephrotic syndrome in mice. PMID- 8422910 TI - Coupling of D2 and D3 dopamine receptors to G-proteins. AB - Recombinant cell lines expressing D2 and D3 dopamine receptor isoforms have been used to study coupling of these receptors to G-proteins from the effects of GTP to reduce agonist binding affinities in agonist/[3H]spiperone competition experiments. D2(short) expressed in Ltk- cells, D2(long) expressed in Ltk- or CHO cells and D3 expressed in CHO cells all showed coupling to the endogenous G proteins of the cells. The detailed agonist binding characteristics indicated that there were differences in receptor/G-protein coupling for the same receptor(D2(long)) expressed in two cell types or for different receptors (D2(short), D2(long)) expressed in the same cell type. PMID- 8422911 TI - Epidermal growth factor and transforming growth factor-alpha can induce neuronal differentiation of rat pheochromocytoma PC12 cells under particular culture conditions. AB - In rat pheochromocytoma PC12 cells, NGF induces neuronal differentiation. Upon stimulation with NGF, Ras is activated to a GTP-bound form, and the activated Ras can induce neuronal differentiation. Recently, we and others observed that epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) can also activate Ras in PC12 cells. This is puzzling since previous reports indicated that EGF stimulates proliferation rather than differentiation in PC12 cells. In this paper, we re-examined the biological effect of EGF and TGF-alpha, and found that these factors can also induce neuronal differentiation under particular culture conditions. Not only the outgrowth of long neurites, but the induction of neurofilament proteins and the metalloprotease transin was also observed in the EGF- and TGF-alpha-stimulated cells. These data clearly indicate that in addition to NGF, EGF and TGF-alpha can also induce the differentiation of PC12 cells under particular conditions. PMID- 8422912 TI - Cyclosporin A suppression of uncoupling in liver mitochondria of ground squirrel during arousal from hibernation. AB - Energy coupling parameters were studied in liver mitochondria of ground squirrel during arousal from hibernation. It was found that such mitochondria become uncoupled during incubation with phosphate in a salt medium. The uncoupling was revealed by respiration rate increase and membrane potential decrease in the presence of oligomycin. Both effects were reversed by addition of cyclosporin A. Under the same in vitro conditions, mitochondria from aroused (active) animals showed no uncoupling but could be uncoupled by addition of palmitate in the cyclosporin A-sensitive fashion. It is proposed that formation of cyclosporin A sensitive pores can be involved in urgent heat production in arousing hibernators. PMID- 8422914 TI - On the localization of FKBP25 in T-lymphocytes. AB - Using polyclonal rabbit antibodies against bovine FKBP25, NEPHGE/SDS-PAGE and Western blotting we demonstrate that the rapamycin-specific immunophilin FKBP25 is present in T-lymphoma Jurkat cells. Subsequent fractionations of the soluble Jurkat cell proteins have revealed that FKBP25 predominantly occurs in the nuclear fraction. FKBP25 has the ability to bind to DNA. The FKBP25/DNA complex can be dissociated in the presence of a high salt concentration. FKBP12, which shares high amino acid sequence homology to the C-terminal domain of FKBP25, has no tendency to bind to DNA. CD-constrained predictions of the secondary structures in FKBP25 suggest that an amphipathic helix-loop-helix occurs in the N terminal part of the protein and may account for its binding to DNA. PMID- 8422913 TI - The tyrosine kinase inhibitors methyl 2,5-dihydroxycinnamate and genistein reduce thrombin-evoked tyrosine phosphorylation and Ca2+ entry in human platelets. AB - Platelet activation is associated with the phosphorylation of a number of platelet proteins at tyrosine residues. The significance of this is unknown. Here we have investigated the effects of two tyrosine kinase inhibitors, methyl 2,5 dihydroxycinnamate and genistein, on thrombin-evoked protein tyrosine phosphorylation and Ca2+ signal generation in fura-2-loaded human platelets. Both compounds inhibited thrombin-evoked tyrosine phosphorylation and reduced the elevation of [Ca2+]i in the presence, but not the absence, of external Ca2+. This suggested a selective inhibition of thrombin-evoked Ca2+ entry but not release from internal stores. Both compounds also reduced thrombin-evoked Mn2+ entry. In contrast, selective blockade of protein kinase C with Ro 31/8220-002 potentiated the thrombin-evoked Ca2+ signal. These data are compatible with a role for protein tyrosine phosphorylation contributing to thrombin-evoked Ca2+ entry in human platelets. PMID- 8422915 TI - A conserved intron in the V-ATPase A subunit genes of plants and algae. AB - Amplification and sequencing of part of the coding regions of the catalytic V type ATPase subunit shows the presence of (at least) two genes in all land plants as well as the conservative insertion of a noncoding sequence. The two genes exhibit a coding region of the same length but differ in the number of nucleotides present in the intron. The latter is surprisingly conserved suggesting the presence of functional constraints on the intron sequences. The findings presented in this report indicate that introns from plants and animals are characterized by different internal structural elements. PMID- 8422916 TI - Apolipoprotein B exhibits phospholipase A1 and phospholipase A2 activities. AB - Low density lipoproteins (LDL) as well as isolated apolipoprotein B (ApoB) have been shown to exhibit phospholipase A2 (PLA2) activity toward phospholipids containing an oxidized or short fatty acyl chain at position 2. Some of these studies employed the fluorescent analogue of phosphatidyl choline (PC), C6-NBD PC, containing NBD-caproic acid (C6-NBD-FA) at position 2 as a substrate, representative of short fatty acyl chains. The release of NBD-caproic acid from position 2 is attributed to PLA2-catalysed hydrolysis. However, this fatty acid can be released also by other enzymatic pathways. In the present study we examined, and ruled out, other enzymatic pathways which may be responsible for the hydrolysis of fatty acids from position 2 of phospholipids. On the other hand, we found that LDL as well as isolated ApoB hydrolyse C6-NBD-FA from both carbon 1 and carbon 2 of these phospholipids, thus exhibiting independent and simultaneous activities of phospholipase A1 and phospholipase A2. PMID- 8422917 TI - Polysialic acids: potential in drug delivery. AB - A number of bacterial polysialic acids were injected intravenously into mice. Half-lives (up to 40 h) in the blood circulation were dependent on the polysialic acid used, increased by deacylation of their phospholipid moiety, decreased with shorter chain derivatives and appeared to be dose independent. A model drug (fluorescein) covalently coupled to a polysialic acid was found to assume the half-life of its carrier. Results suggest that intact or deacylated polysialic acids and shorter chain derivatives can be used to augment the half-lives of drugs, small peptides, proteins and drug delivery systems in the blood circulation, thus prolonging their pharmacological action. PMID- 8422918 TI - Conformational junctions between left-handed DNA in (dA-dT)16 and contiguous B DNA in a supercoiled plasmid contain chemically reactive bases. AB - Alternating adenine-thymine sequences in supercoiled DNA may undergo a transition to the left-handed Z-conformation in the presence of Ni2+ ions and high Na+ concentrations [(1989) FEBS Lett. 243, 313-317]. In this work we have studied the junctions between B- and Z-conformations in a supercoiled plasmid containing a (dA-dT)16 insert, by means of chemical probing. We observed enhanced reactivity of bases at both ends of the alternating tract to chloro- and bromoacetaldehyde. The degree of chemical reactivity was found to increase with the level of negative supercoiling. Only individual bases were observed to be reactive in the B-Z junctions, consistent with tightly localized interfacial regions. PMID- 8422919 TI - Temporal nature of the promoter and not relative strength determines the expression of an extensively processed protein in a baculovirus system. AB - We demonstrate that the expression of extensively modified and secreted heterologous proteins synthesized in the baculovirus expression vector system (BEVS) depends on the temporal nature of the promoter transcribing the foreign gene. The beta subunit of the human chorionic gonadotropin, an extensively modified secretory glycoprotein hormone was expressed under the transcriptional control of the AcNPV basic protein gene promoter (MP) and the polyhedrin gene promoter (POL), respectively. MP, activated late in the infection cycle, is a weaker promoter when compared to the stronger very late POL promoter. Levels of secretion, immunoreactivity and bioactivity of recombinant proteins, beta hCGMP and beta hCGPOL synthesized under control of the MP and POL promoter were compared. Secretion of beta h CGMP was relatively higher. Enzymatic analysis revealed that the synthesized protein was sialylated. Receptor binding assays and testosterone stimulation assays in a mouse Leydig cell system demonstrated that on a unit protein basis, beta hCGMP was biologically more active than beta hCGPOL. PMID- 8422920 TI - Primary structure of crayfish visual pigment deduced from cDNA. AB - The primary structure of opsin of the crayfish Procambarus clarkii has been deduced from the cDNA sequence. The opsin is composed of 376 amino acid residues including all the conservative residues characteristic of other members of the rhodopsin family. Comparison of sequences of all known opsins reveals that the major Drosophila rhodopsin is more similar to the crayfish rhodopsin than to the Drosophila UV-sensitive pigments. The phylogenetic trees of invertebrate opsins are constructed. PMID- 8422921 TI - Structural alignment of globins, phycocyanins and colicin A. AB - A database search employing a novel algorithm for protein structure comparison by alignment of distance matrices has revealed a striking resemblance between the tertiary structures of the bacterial toxin colicin A and globins. The globin-like domain in colicin A contains all elements essential for the toxin's lethal ionophoric activity. The structural similarity between colicin A and globins is comparable to that between globins and phycocyanins. This suggests that these three protein families, which have unrelated sequences and different functional contexts, are an example of physical convergence to a stable folding motif, the three-on-three helical sandwich. PMID- 8422922 TI - Na(+)-H+ exchange in sheep parotid endpieces. Apparent insensitivity to amiloride. AB - We have used microspectrofluorimetry with the pH-sensitive dye, BCECF, to examine the control of intracellular pH in the secretory endpieces of the sheep parotid gland. Unstimulated endpieces in HCO3(-)-free media have a cytosolic pH of 7.5 +/ 0.03 (n = 69) which is maintained by a Na(+)-dependent proton extrusion process that can be partially supported by Li+ but not by Cs+, and is not affected by changes in extracellular Cl-, HCO3- or K+. It is not blocked by SITS or DIDS, which inhibit Na(+)-(n)HCO3- co-transport and CL(-)-HCO3- exchange, nor is it sensitive to the amiloride analogs, MIA and EIPA, which inhibit Na(+)-H+ exchangers, although very high concentrations of amiloride itself (1 mmol/l) have a (probably non-specific) inhibitory effect. It seems likely that sheep parotid secretory endpieces do contain a Na(+)-H+ exchanger that drives secretion of a HCO3(-)-rich juice, and that its insensitivity to amiloride and its analogs explains why these drugs do not block fluid secretion by the intact sheep parotid gland. PMID- 8422923 TI - Genomic DNA structure of a gene encoding cytosolic ascorbate peroxidase from Arabidopsis thaliana. AB - A genomic DNA clone encoding cytosolic ascorbate peroxidase was isolated from a genomic library of Arabidopsis thaliana, using a cDNA for the enzyme as a probe. Nucleotide sequence and primer extension analyses of this gene (APX1) revealed nine exons split by eight introns, one of which is inserted in the 5' untranslated region. The exon/intron organization of the APX1 gene differs from that of the guaiacol peroxidase genes. PMID- 8422924 TI - The rat delta-1 and delta-2 subunits extend the excitatory amino acid receptor family. AB - We have characterized a second member (delta-2) of a new class of subunits for the ligand-gated excitatory amino acid receptor superfamily. The sequence of delta-2 exhibits an average identity of 25% and 18.5% to the non-NMDA and NMDA receptor subunits, respectively. The rat delta-2 gene is expressed predominantly in Purkinje cells of the cerebellum whereas only low levels of delta-1 transcripts are found in the adult brain. However, delta-1 gene expression undergoes a pronounced developmental peak, with particularly high mRNA levels in the caudate putamen of late embryonic/early postnatal stages. PMID- 8422925 TI - A peptide corresponding to the N-terminal 13 residues of T4 lysozyme forms an alpha-helix. AB - Solid-phase methods have been used to synthesize LYS(1-13), a peptide corresponding to the first 13 residues of T4 lysozyme. 2D 1H NMR techniques were used to investigate its solution structure in the presence of SDS micelles. The identification of numerous medium-range NOESY crosspeaks and several slowly exchanging NH protons indicated the presence of an alpha-helical structure. This was confirmed by simulated annealing calculations performed using XPLOR. PMID- 8422926 TI - In vitro expression and activity of lycopene cyclase and beta-carotene hydroxylase from Erwinia herbicola. AB - The cyclisation of lycopene to beta-carotene and the hydroxylation of beta carotene to zeaxanthin are common enzymatic steps in the biosynthesis of carotenoids in a wide range of bacteria, fungi, and plants. We have individually expressed in E. coli the two genes coding for these enzymatic steps in Erwinia herbicola. The cyclase and hydroxylase enzymes have apparent molecular weights of 43 kDa and 22 kDa, respectively, as determined by SDS-PAGE. Hydroxylase in vitro activity was obtained only in the cytoplasmic fraction. Cyclase also demonstrated enzyme activity in a crude cell-free lysate, although to a lesser extent. PMID- 8422927 TI - Stoichiometry of the binding of chromosomal protein MC1 from the archaebacterium, Methanosarcina spp. CHTI55, to DNA. AB - We have investigated the binding stoichiometry of the chromosomal MC1 protein on DNA using the gel retardation technique. Analysis of the distribution of the complex containing 0, 1, 2, 3 ... bound proteins shows that the protein MC1 interacts with the DNA as a monomer. Binding experiments with short DNA fragments of various lengths shows that the site size is 11 bp in length. These results are compared to those obtained with other chromosomal proteins including HU protein. PMID- 8422928 TI - Cloning and sequence analysis of cDNA for the Ca(2+)-activated photoprotein, clytin. AB - Clytin is a member of the aequorin family of photoproteins. It is made up of 189 amino acid residues, contains 3 Ca(2+)-binding sites, and shows 62% homology in amino acid residues to those in aequorin. The cysteine, tryptophan, and histidine residues, and the C-terminal proline, that are conserved in aequorin and clytin may be involved in the Ca(2+)-activated bioluminescence of the two proteins. Clytin may also prove useful in the determination of Ca2+. PMID- 8422929 TI - A ubiquityl-calmodulin synthetase that effectively recognizes the Ca(2+)-free form of calmodulin. AB - Ubiquityl-calmodulin synthetase (uCaM-synthetase) activity as detected in reticulocyte lysate and the crude extracts of rabbit tissues [FEBS Lett. 294 (1991) 229-233] has been well characterized as being essentially Ca(2+)-dependent (-Ca2+/+Ca2+ activity ratio: 0.15-0.2). However, during the purification of this enzyme on ubiquitin-Sepharose the Ca(2+)-dependent activity is lost and an essentially Ca(2+)-independent enzyme (-Ca2+/+Ca2+ activity ratio: 1.0-1.5) is obtained which was purified 90-fold (uCaM-Syn F1) to a final specific activity of 0.32 pkat/mg. During the purification procedure a second protein factor (uCaM-Syn F2) was isolated that has no catalytic activity by itself but restores Ca2+ dependence to the uCaM-Syn F1 fraction (-Ca2+/+Ca2+ activity ratio: 0.1) and enhances the catalytic activity in uCaM-Syn F1 in the presence of Ca2+ over 40 fold. It is concluded that several (possibly interdependent) forms of uCaM synthetase exist which display different substrate specificities for calmodulin. PMID- 8422930 TI - Role of a serum phospholipase A1 in the phosphatidylserine-induced T cell inhibition. AB - We have previously shown that unsaturated phosphatidylserines inhibit mitogen induced T cell activation. We now report that the inhibitory action requires a protein present in bovine and human serum. Partial purification and phospholipase assay show that this protein has phospholipase A activity on phosphatidylserine but not phosphatidylethanolamine, phosphatidylcholine and phosphatidylinositol. In short incubations (1-3 h)2-acyl lysophosphatidylserine is produced but in longer incubations the cis-unsaturated fatty acid is also released. Experiments on peripheral blood mononuclear cells indicate that the unsaturated fatty acid becomes the main responsible for the PS-induced inhibition and that 2-acyl lysophosphatidylserine enhances the inhibitory effect of fatty acid. PMID- 8422931 TI - Comment to Knoop et al. (1990) FEBS Letters 267, 9-12, toxin B of Clostridium difficile does not have enolase activity. PMID- 8422932 TI - Localization of Kex2-like processing endoproteases, furin and PC4, within mouse testis by in situ hybridization. AB - By in situ hybridization analysis, we show here the localization of furin and PC4, which are both members of a growing family of endoproteases structurally related to the yeast precursor processing protease Kex2, within mouse testis. Furin transcript was detected in both germ and somatic cells, while PC4 transcript was found only in round spermatids. Proenkephalin transcript was also localized in round spermatids. These observations suggest that, within testis, PC4 is involved in processing of peptide precursors such as proenkephalin and may play a role in regulation of sperm maturation, while furin may serve as a more general processing endoprotease. PMID- 8422933 TI - Gluten exorphin C. A novel opioid peptide derived from wheat gluten. AB - A novel opioid peptide, Tyr-Pro-Ile-Ser-Leu, was isolated from the pepsin-trypsin chymotrypsin digest of wheat gluten. Its IC50 values were 40 microM and 13.5 microM in the GPI and MVD assays, respectively. This peptide was named gluten exorphin C. Gluten exorphin C had a structure quite different from any of the endogenous and exogenous opioid peptides ever reported in that the N terminal Tyr was the only aromatic amino acid. The analogs containing Tyr-Pro-X-Ser-Leu were synthesized to study its structure-activity relationship. Peptides in which X was an aromatic amino acid or an aliphatic hydrophobic amino acid had opioid activity. PMID- 8422934 TI - Developmental changes in the expression of HMG 2a protein. AB - The levels of HMG 2a chromosomal protein and its mRNA change during the post hatched development of chicks were investigated. The contents of both HMG 2a and 2b proteins of liver, heart, brain, muscle and gizzard were abundant in the newly hatched chicks but their contents decreased significantly in those tissues of the 70-day-old chicks. The HMG 2a mRNA levels of liver, heart and brain in 70-day-old chick decreased to about 40% of those mRNA in the newly hatched chicks while the HMG 2a mRNA levels of muscle and gizzard in the 70-day-old chicks increased 5- and 3-fold, respectively. These results suggest that the decrease in the HMG 2a protein contents of the muscle and gizzard in the 70-day-old chicks may be largely due to the stimulation of HMG 2a protein degradation or the reduction of HMG 2a mRNA translation. PMID- 8422935 TI - Sequence similarities in (alpha/beta)8-barrel enzymes revealed by conserved regions of alpha-amylase. AB - The parallel (alpha/beta)8-barrel is a frequently occurring protein-folding motif. Although the arrangement of secondary structural elements along the barrel is very similar in different (alpha/beta)8-barrel enzymes, there is a very low mutual amino acid sequence homology among the enzymes, contributing in part to the hazy view of their evolution. Here an approach to identifying at least the rough of evolutionarily conserved (alpha/beta)8-barrel sequence is presented. Based on the idea that highly conserved sequence regions of a particular enzyme should be more or less conserved in the sequences of the other evolutionary related enzymes, five sequence similarities of ten different (alpha/beta)8-barrel enzymes were revealed, using the five conserved regions of the amino acid sequence of the alpha-amylase (alpha/beta)8-barrel as the templates. PMID- 8422936 TI - Prevention of phospholipase-C induced aggregation of low density lipoprotein by amphipathic apolipoproteins. AB - Phospholipase C (PL-C) digestion of human low density lipoprotein (LDL) results in hydrolytic cleavage of the phosphocholine head group of phosphatidylcholine, thereby generating diacylglycerol. Loss of amphiphillic surface lipids and/or accumulation of diacylglycerol causes LDL samples to develop turbidity. Examination of PL-C treated LDL by electron microscopy revealed a progressive aggregation of LDL as a function of phosphatidylcholine hydrolysis: fused particles, clusters, and multiple stacked aggregates were observed. Lipid analysis of untreated and aggregated LDL confirmed that the phosphatidylcholine content of the latter had decreased with a corresponding increase in diacylglycerol. It is likely that phospholipolysis created hydrophobic gaps within the surface monolayer of LDL, thereby inducing LDL fusion and aggregation. When amphipathic alpha-helix-containing apolipoproteins, such as human apoA-I or Manduca sexta apolipophorin III (apoLp-III) were present, PL-C treated LDL did not aggregate. Compositional analysis of apolipoprotein-containing PL-C LDL showed that phospholipolysis was not affected by the presence of apolipoproteins. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of lipoproteins re isolated following incubation with PL-C revealed an association of apoA-I or apoLp-III with PL-C digested LDL. Electron microscopy showed no major morphological differences between native LDL and apoprotein stabilized PL-C treated LDL and the average particle diameter of apoA-I stabilized PL-C LDL was 22.5 +/- 2.2 nm versus 22.8 +/- 1.6 nm for control LDL. Incubation of tritium labeled apoLp-III with LDL and PL-C demonstrated that association of apoLp-III with PL-C LDL correlated with the extent of phospholipid hydrolysis, the apolipoproteins apparently being recruited to compensate for the increased hydrophobic surface created by conversion of phosphatidylcholine into diacylglycerol. The results suggest that transient association of amphipathic apolipoproteins with damaged or unstable LDL may provide a mechanism to obviate formation of atherogenic LDL aggregates in vivo. PMID- 8422937 TI - High levels of profilin suppress the lethality caused by overproduction of actin in yeast cells. AB - Overproduction of actin is lethal to yeast cells. In contrast, overexpression of the profilin gene, PFY1, encoding an actin-binding protein, leads to no very obvious phenotype. Interestingly, profilin overproduction can compensate for the deleterious effects of too much actin in a profilin concentration-dependent manner. Our results, thus, document that actin and profilin interact in vivo. Immunofluorescence studies suggest that suppression works by reducing actin assembly. We observed, however, that even massive overproduction of profilin fails to fully restore the wild-type phenotype (e.g. the wild-type appearance of the actin microfilament system). This may indicate that actin monomer sequestration is not the only mechanism by which the balance of actin polymerization is controlled. PMID- 8422938 TI - The C-terminal tripeptide of glycosomal phosphoglycerate kinase is both necessary and sufficient for import into the glycosomes of Trypanosoma brucei. AB - Glycosomal phosphoglycerate kinase (gPGK) of Trypanosoma brucei differs from the cytoplasmic isozyme (cPGK) in its higher isoelectric point characterized by clusters of positive charges along the polypeptide chain, and a 20 amino acid C terminal extension ending in serine-serine-leucine (SSL). While a C-terminal SSL tripeptide is apparently not capable of directing luciferase to the peroxisomes in mammalian cells [J. Cell Biol. 108 (1989), 1657-1664], we show here that it is sufficient for the import of luciferase as well as an unrelated protein, beta glucuronidase, into the glycosomes of T. brucei, as determined by immunoelectron microscopy. The analysis of luciferase-gPGK fusion proteins indicates that the only targeting signal for import of gPGK into the glycosome resides in this C terminal SSL sequence. PMID- 8422939 TI - Synthesis and binding characteristics of two sulfhydryl-reactive probes for vasopressin receptors. AB - The present study describes the synthesis and receptor binding affinities of the sulfhydryl-reactive vasopressin analogs deamino[Dab(N delta-N-maleoyl-beta-alanin e)4]AVP (1a) and deamino[Lys(N epsilon-N-maleoyl-beta-alanine)8VP (2a). The analogs were obtained by introducing the sulfhydryl-reactive maleoyl-beta-analyl group at the delta-amino group of Dab4 in deamino[Dab4]AVP (1) and at the epsilon amino group of Lys8 in deamino[Lys8]VP (2), which were synthesized by the solid phase method. Furthermore, the analog modified at Dab4 was prepared as tritium labeled compound (1b) after catalytic iodine tritium exchange at Tyr2 in deamino[Dab4]AVP. The sulfhydryl-reactive vasopressin analogs retained high binding affinity for the V2 vasopressin receptor in membranes derived from bovine kidney inner medulla. Apparent dissociation constants Kd of 45 nM (compound 1a) and 15 nM (compound 2a) were determined. Incubation of the ligand receptor complexes at pH 5.5 resulted in dissociation of the sulfhydryl-reactive vasopressin analogs from the V2 receptor. No indications of a covalent reaction between analogs 1a, 2a and 1b and sulfhydryl groups in or close to the hormone binding site of the V2 receptor were found. PMID- 8422940 TI - Cloning of the rat heart Na(+)-Ca2+ exchanger and its functional expression in HeLa cells. AB - A functional rat heart Na(+)-Ca2+ exchanger gene has been obtained by splicing and ligating two partially overlapping clones isolated from a rat heart lambda ZAP cDNA library. The deduced primary structure of the protein encoded by the open reading frame corresponds to 971 amino acids, that can be organized into 12 transmembrane helices. The cloned gene was functionally expressed in HeLa cells. Maximal expression was detected 18 h after transfection, after which transport activity rapidly declined. The electrogenic properties of the cloned transporter were demonstrated following reconstitution of the expressed exchanger protein into a tightly sealed phospholipid membrane. PMID- 8422941 TI - Calcium homeostasis in yeast cells exposed to high concentrations of calcium. Roles of vacuolar H(+)-ATPase and cellular ATP. AB - Cytosolic Ca2+ concentrations ([Ca2+]i) were determined in haploid and diploid cells of Saccharomyces cerevisiae, loaded with indo-1 and exposed to media containing a range of Ca2+ concentrations. [Ca2+]i homeostasis was maintained at the 100-150 nM level in cells which were pre-incubated with glucose and exposed to 0.1 microM-10 mM Ca2+ in the medium. Slightly higher levels of [Ca2+]i were determined in cells exposed to 50 mM Ca2+. Pre-incubation with metabolic inhibitors instead of glucose caused a reduction in cellular ATP levels and an impaired [Ca2+]i homeostasis; [Ca2+]i reached 800 nM in cells exposed to 10 mM CaCl2. Cells of the delta vma4 mutant strain, with no functional vacuolar H(+) ATPase, had elevated levels of [Ca2+]i, reaching 1.8 microM when pre-incubated with glucose and exposed to 10 mM CaCl2. Higher levels of [Ca2+]i were measured in the mutant cells which were pre-incubated with metabolic inhibitors. This result indicates the central role of the vacuoles in maintaining [Ca2+]i homeostasis and suggests the presence of an additional non-vacuolar ATP-requiring mechanism which contributes to keeping [Ca2+]i at low levels. PMID- 8422942 TI - Early induction of Na(+)-dependent uridine uptake in the regenerating rat liver. AB - Na(+)-dependent uridine transport into liver plasma membrane vesicles from partially hepatectomized and sham-operated rats was studied. Preparations purified 6 h after 70% hepatectomy exhibited an increased Vmax of uridine uptake (3.7 vs. 1.4 pmol/mg prot/3 s) without any change in Km (6 microM). Incubation of the vesicles in the presence of monensin decreased uridine uptake although the differences between both experimental groups remained identical. It is concluded that uridine transport is induced early after partial hepatectomy by a mechanism which does not involve changes in the transmembrane Na+ gradient. This is the first evidence in favor of modulation of nucleoside transport into liver cells. PMID- 8422943 TI - N-terminal amino acid sequences of the subunits of the Na(+)-translocating F1F0 ATPase from Propionigenium modestum. AB - We report here the N-terminal protein sequences of the subunits of the ATPase from Propionigenium modestum. Subunits c, b, delta, alpha and beta start with an N-terminal methionine residue, the gamma and epsilon subunits have an alanine N terminus, from which N-formylmethionine was hydrolyzed by posttranslational modification, and subunit a contains a blocked N-terminus. Each of the N-terminal sequences exactly matches a portion of the DNA sequence in the gene encoding the respective subunit protein on the unc operon. Thus, the exact translational start for each subunit protein can be identified and the primary structures of the protein transcripts can be clearly defined. Based on these data the putative size of the open reading frame that was envisaged from the DNA sequence had to be revised for the alpha and delta subunits. PMID- 8422944 TI - Chloroplast envelope protein encoded by chloroplast genome. AB - The gene product of an open reading frame of chloroplast genome, ORF 231 in pea, was immunochemically detected in chloroplast and etioplast envelopes. This is the first protein of a Chloroplast Envelope Membrane encoded by a chloroplast genome. It was named CEM A and the gene, cem A. CEM A is an acidic protein having an apparent molecular mass of 34 kDa on SDS-PAGE, and a minor component detected in the fractionated inner envelope. PMID- 8422945 TI - Thromboxane A2 analogue U 46619 enhances tumour cell proliferation in HeLa cells via specific receptors which are apparently distinct from TXA2 receptors on human platelets. AB - In this study, we have demonstrated for the first time by using U 46619, a stable analogue of thromboxane A2 (TXA2), that TXA2 exerts a cell proliferative effect on HeLa cells which is mediated by specific TXA2 receptors, inasmuch as the cell proliferation could be dose-dependently suppressed by TXA2 receptor antagonist BM 13177. The investigation of the phospholipase C pathway by U 46619 and prostaglandin H2 (PGH2) in the presence and absence of BM 13177 in cells with or without pertussis toxin pretreatment, as well as radioligand receptor binding studies, revealed that, in contrast to TXA2 receptors on human platelets, where TXA2 and PGH2 share the same receptor binding sites, HeLa cells possess distinct receptors for TXA2 and PGH2. PMID- 8422946 TI - The 'non-exchangeable' nucleotides of F1-F0ATP synthase. Cofactors in hydrolysis? AB - The F1-F0 ATP synthase bears 6 nucleotide binding sites, only 3 of which turn over during catalysis. The remaining 3 are occupied by slowly exchanging ATP in vivo, although at least 1 molecule is generally lost on isolation of the enzyme in the absence of nucleotide. It is proposed that the function of the slowly exchanging (NC) nucleotides is to participate in catalysis, the terminal phosphate of the bound ATP acting as an acid catalyst in the cleavage/synthesis of the phosphate anhydride bond in the catalytic sites. Such a role has been demonstrated for the bound pyridoxal phosphate moiety in glycogen phosphorylase. Evidence is presented that (i) the NC nucleotide spans the interface between an alpha subunit and its partner beta, interacting near the catalytic binding site on beta; (ii) the phosphate moieties of the catalyzed and NC nucleotide are close in space; and (iii) occupation of the NC nucleotide sites promotes ATP hydrolysis by F1 or its subfragments. All of these findings are required by the proposed mechanism. Relationships between phosphorylase and F1 structures are discussed. PMID- 8422947 TI - FAD-linked glycerophosphate dehydrogenase deficiency in pancreatic islets of mice with hereditary diabetes. AB - The mitochondrial enzyme FAD-linked glycerophosphate dehydrogenase plays a key role in the glucose-sensing device of the insulin-producing pancreatic B-cell. Its activity was found to be decreased in islet, but not liver, homogenates of BL/Ks-db/db mice, in which diabetes mellitus represents an inherited disease. The decreased activity of FAD-linked glycerophosphate dehydrogenase contrasted with a normal activity of glutamate dehydrogenase and 2-ketoglutarate dehydrogenase in the islets of db/db mice. It is proposed that a site-specific defect of FAD linked glycerophosphate dehydrogenase in the pancreatic B-cell might represent a far-from-uncommon causal or contributing factor in the pathogenesis of non insulin-dependent diabetes mellitus. PMID- 8422948 TI - Functional expression of 5-HT1c receptor cDNA in COS 7 cells and its influence on protein kinase C. AB - Two subtypes of receptors for serotonin (5-hydroxytryptamine; 5-HT) are known to stimulate inositol (1,4,5)-trisphosphate production, the 5-HT1c and 5-HT2 receptors. In this study we investigated the ability of 5-HT1c receptors, transiently expressed in COS 7 cells, to functionally interact with protein kinase C-alpha, the indigenous (phorbol ester-responsive) isoform of the enzyme in those cells. Serotonin caused translocation of the [3H]phorbol 12,13 dibutyrate (PDBu) binding site of PKC-alpha from the cytosolic to the membrane fraction in a Ca(2+)-dependent manner which was prevented by the 5-HT1c receptor antagonist mianserin. The lipid activators of PKC, PDBu and 1,2-dioctanoyl-sn glycerol (DOG) also caused translocation, but through a mechanism apparently quite independent of Ca2+. PMID- 8422949 TI - A new family of basic cysteine-rich plant antifungal proteins from Brassicaceae species. AB - Out of seeds of 4 Brassicaceae species, 7 antifungal proteins were isolated which are nearly identical to 2 previously characterized radish seed antifungal proteins. These basic proteins, multimers of a 5 kDa polypeptide, specifically inhibit fungal growth. One of the antifungal proteins has decreased antifungal activity and an increased antibacterial activity. In addition, the previously described antifungal activity of the radish seed 2S albumins was extended to the 2S albumins of the seeds of the 4 other Brassicaceae species. A 2S albumin-like trypsin-inhibitor from barley seeds was found to have much less activity against fungi. PMID- 8422950 TI - Conserved secondary structures in the ITS2 of trematode pre-rRNA. AB - Biological functions of transcribed spacer regions in eucaryotic pre-rRNAs remain elusive so far. Utilization of the comparative approach to analyse their secondary structure has been impeded by the extensive sequence divergence observed among most of the specimens available to date. However, we have taken advantage of the recent derivation of a set of largely similar sequences for different schistosome species to look for the presence of constrained secondary structures within the internally transcribed spacer 2 (ITS2). We show that several common features of secondary structure are shared by these species despite sequence variation, with some of them supported by compensatory changes, suggesting a significant role of ITS2 as an RNA domain during ribosome biogenesis. PMID- 8422951 TI - Expression of highly active sex-inducing pheromone of Volvox carteri f. nagariensis in a mammalian cell system. AB - A cDNA fragment coding for the sex-inducing glycoprotein of Volvox carteri f. nagariensis was expressed in a mammalian cell system (baby hamster kidney (BHK) cells). The transfection product exhibited a specific biological activity intermediate between the natural pheromone of the strains Volvox carteri f. nagariensis and Volvox carteri f. weismannia. Immunoblot analysis showed that the sex-inducing activity was expressed as a set of three iso-glycoproteins (35, 34 and 31 kDa). PMID- 8422952 TI - In vitro complex formation between cholesterol and alpha 1-proteinase inhibitor. AB - The in vitro interaction between human alpha 1-proteinase inhibitor (alpha 1-PI) and cholesterol was studied with electrophoretic and gel chromatographic methods. The addition of cholesterol (from 1 to 20 mol/mol alpha 1-PI) at 37 degrees C resulted in retarded electrophoretic mobility of alpha 1-PI towards the anode, diminished immunoreactivity and antiproteinase activity. At a molar ratio of 2:1 (cholesterol/alpha 1-PI), antitryptic activity was reduced by 15% but antielastase activity by 50%. At this ratio the gel filtration alpha 1-PI peak appeared at 67 kDa, as compared to 52 kDa for native alpha 1-PI. No size difference was noted on SDS-PAGE. These results suggest the occurrence of noncovalent complex formation between cholesterol and alpha 1-PI in vitro. PMID- 8422953 TI - Adenosine analogs inhibit the guanine-7-methylation of mRNA cap structures. AB - The adenosine analogs neplanocin A and deazaneplanocin A were observed to inhibit the in vivo guanine-7-methylation of mRNA cap structure using a new assay for hypomethylated RNA. Treatment of cultured mammalian cells with these adenosine analogs resulted in the same extent of hypomethylation of cap structure as did ethionine injection in mice. Neplanocin A and its non-metabolizable analog 3 deazaneplanocin A show the same maximal level of inhibition of methylation suggesting that these adenosine analogs exert their effects by elevating S adenosylhomocysteine levels rather than by conversion to other inhibitory compounds. PMID- 8422954 TI - Effects of tyrosine kinase inhibitors on protein kinase-independent systems. AB - Tyrosine kinase inhibitors have been widely used to probe the role of tyrosine phosphorylation in cellular signalling. These inhibitors exhibit an apparent specificity for tyrosine kinases over the serine/threonine kinases but little is known about their effects on other enzymes or biological systems. We demonstrate that genistein, erbstatin and alpha-cyanocinnamamides (tyrphostins) have inhibitory effects on fatty acid synthesis, lactate transport, mitochondrial oxidative phosphorylation and aldehyde dehydrogenase. We propose, therefore, that results obtained using tyrosine kinase inhibitors should be interpreted with caution, particularly if used at concentrations sufficient to inhibit these non protein kinase-dependent events. PMID- 8422955 TI - Forskolin stimulates porcine sperm capacitation by increasing calcium uptake. AB - Using the fluorescent calcium indicator fura-2, forskolin was found to dose dependently cause an immediate increase in the concentration of intracellular free calcium of porcine cauda epididymal sperm. This stimulatory effect of forskolin is due to the enhancement of Ca2+ uptake by the verapamil-sensitive transporter on the sperm plasma membrane and results in the promotion of the sperm capacitation and subsequent acrosome reaction. PMID- 8422956 TI - In vitro selection and evolution of RNA: applications for catalytic RNA, molecular recognition, and drug discovery. AB - In vitro selection and in vitro evolution methods represent powerful tools for isolating functional RNA molecules, and are proving to have wide applications in biology. Selection in the absence of living cells is possible because some RNA molecules possess a selectable "phenotype" (catalytic activity or ligand binding) as well as a "genotype" (nucleotide sequence). This review discusses the basic principles of in vitro selection technology and the application of these methods to isolate RNA molecules with interesting and novel properties. Selection techniques have been used to analyze the structure and function of catalytic RNA molecules (ribozymes), and to isolate novel catalytic structures not found in nature. They are also useful for studying protein-RNA interactions and for isolating RNA molecules that bind specifically to peptides and other ligands. The isolation of RNA molecules with new binding functionalities (aptamers) for both large and small molecules has exciting potential for discovery of new drugs and diagnostic reagents. PMID- 8422957 TI - Ribosomal RNA: a key to phylogeny. AB - As molecular phylogeny increasingly shapes our understanding of organismal relationships, no molecule has been applied to more questions than have ribosomal RNAs. We review this role of the rRNAs and some of the insights that have been gained from them. We also offer some of the practical considerations in extracting the phylogenetic information from the sequences. Finally, we stress the importance of comparing results from multiple molecules, both as a method for testing the overall reliability of the organismal phylogeny and as a method for more broadly exploring the history of the genome. PMID- 8422958 TI - Point and compensation mutations to evaluate essential stem structures of genomic HDV ribozyme. AB - In elucidating the part played by the essential stem structures (I, II, and III) in the self-cleavage activity of genomic HDV ribozyme, several point and compensation variants were constructed on pseudoknot-like structure by site directed mutagenesis. The self-cleavage activities of these variants indicated that stems I and III were essential for the activity by forming Watson-Crick base pairs. On the other hand, disruption of A704:U767 had little influence on the cleavage activity, indicating that it is not essential in forming an active structure. Also, our V1 nuclease probing studies showed that the A704U and HDV88 variants have a sensitivity similar to the nuclease, and major cuts are visible in the stem I and stem II regions. Thus, stem I and stem II regions are maintained together with stem III regions in both molecules. These results and our earlier site-directed mutagenesis studies strongly support a pseudoknot-like structure for the genomic HDV ribozyme. PMID- 8422959 TI - Modification interference analysis of a self-cleaving RNA from hepatitis delta virus. AB - A chemical modification-interference assay was used to evaluate the sequence requirements for self-cleavage of a 73-nucleotide self-cleaving RNA from the genomic hepatitis delta virus (HDV). Twenty-two nucleotides were categorized as individually essential for self-cleavage, shown by loss of activity when modified. All of these required nucleotides fell within 38 nucleotides downstream of the cleavage site, suggesting an essential structural or functional role for this region. Lesser effects were seen for nucleotides further 3' of the cleavage site, and a small number of nucleotides had a negligible effect on the extent of self-cleavage when modified. Several modifications increased the extent of self cleavage, suggesting these nucleotides may act to inhibit the reaction when unmodified. The functional requirements for certain nucleotides are discussed in the light of structural probing data and conventional mutational analysis available for other HDV RNAs. PMID- 8422960 TI - Theoretical analyses on the role of Mg2+ ions in ribozyme reactions. AB - To elucidate the role of the Mg2+ ion in ribozyme reactions, we carried out ab initio molecular orbital investigations on dianionic trimethoxyphosphorane A and its Mg2+ complex (overall a neutral molecule) as a model system for the reaction center of Tetrahymena-type ribozyme. Although dianionic oxyphosphorane A concentrates its negative charges on the equatorial phosphoryl oxygens, the coordination of the Mg2+ ion between these two oxygens is unlikely. Geometry optimizations of the complex and the electrostatic potential of A both suggest that Mg2+ coordination preferably occurs in the region between the axial oxygen and the equatorial phosphoryl oxygen. The considerations of electrostatic potential rationalize the geometries of carboxylate-metal and phosphate-metal interactions extracted from the Cambridge Structural Database as well. Consequently, the Mg2+ ion at the active site of Tetrahymena-type ribozyme most likely lies in the regions between the axial and equatorial oxygens. The axial equatorial coordinations of Mg2+ ions conceivably increase the electronegativities of the axial oxygens and facilitate cleavage of the phosphodiester bond located at the junction of the intron and the exon. It is thus likely that the Mg2+ ions play the key role in the phosphodiester cleavage reactions mediated by ribozymes. PMID- 8422961 TI - Multiple exoribonucleases are required for the 3' processing of Escherichia coli tRNA precursors in vivo. AB - Our knowledge of the 3' processing of tRNA precursors is severely limited. Although six exoribonucleases able to act on Escherichia coli tRNA precursors in vitro have been identified, their involvement in tRNA maturation in vivo has not been demonstrated. Here we show, using a wide range of multiple RNase-deficient strains and a quantitative suppression assay, that at least five of these enzymes -RNase II, RNase D, RNase BN, RNase T, and RNase PH--can participate in the synthesis of functional tRNA(Tyr)su+3 in vivo. Moreover, any one of the five RNases is sufficient to allow tRNA processing to proceed although with varying effectiveness. Examination of the level of aminoacylation of tRNA isolated from RNase-deficient strains suggested that tRNA precursors accumulate in the most defective cells. These data indicate that exoribonucleases are required for tRNA maturation in vivo and that there is a high degree of functional overlap among the enzymes. These studies contribute to the identification of all the enzymes necessary for defining the complete processing pathway for E. coli tRNA precursors. PMID- 8422962 TI - Group I and group II introns. AB - Group I and group II introns are two types of RNA enzymes, ribozymes, that catalyze their own splicing by different mechanisms. In this review, we summarize current information about the structures of group I and group II introns, their RNA-catalyzed reactions, the facilitation of RNA-catalyzed splicing by protein factors, and the ability of the introns to function as mobile elements. The RNA based enzymatic reactions and intron mobility provide a framework for considering the role of primordial catalytic RNAs in evolution and the origin of introns in higher organisms. PMID- 8422963 TI - Clustering of modified nucleotides at the functional center of bacterial ribosomal RNA. AB - An aryl trifluoromethyl diazirine photoreactive derivative was attached to the 2 thiocytidine residue at position 32 of tRNA(IArg) and this derivatized tRNA was bound to Escherichia coli 70S ribosomes. After irradiation at 350 nm the site of cross-linking to the 16S RNA was analyzed by our standard procedures and found to lie within the secondary structural element comprising bases 956-983; this region contains two modified nucleotides at positions 966 and 967. Similarly, an aryl azido photoreactive derivative was attached to the phenylalanine residue of Phe tRNA(Phe), and the derivatized aminoacyl tRNA was bound to the ribosome either at the A- or the P-site. In both cases, after irradiation at 250 nm, the cross-link site was localized to position 2439 of the 23S RNA; in the secondary structure of the latter the neighboring nucleotide 2442 is base-paired to a modified nucleotide at position 2069. Taken together with other cross-linking data, these results now directly implicate a total of 27 out of the 29 modified nucleotides in E. coli 16S and 23S RNA as lying within or close to the functional center of the ribosome. PMID- 8422964 TI - Energy cost of proofreading in vivo: the charging of methionine tRNAs in Escherichia coli. AB - Previous in vitro work has shown that Escherichia coli methionyl-tRNA synthetase has a limited ability to discriminate against cognate methionine in the editing site designed for noncognate homocysteine. As a result, a small fraction of the correct product Met-tRNA is deacylated with the formation of a cyclic sulfonium compound, S-methyl-homocysteine thiolactone. This is exploited here to estimate energy costs associated with the destruction of a correct product by methionyl tRNA synthetase in bacterial cells. In vivo measurements of S-methyl-homocysteine thiolactone indicate that in Escherichia coli 3.3 molecules of Met-tRNA are destroyed by deacylation per 10,000 molecules of Met-tRNA successfully transferring methionine to protein. This number of destroyed molecules of a correct product, Met-tRNA, is 30 times lower than the number of destroyed molecules of an incorrect product, homocysteinyl adenylate. Thus, most of the energy cost of proofreading in vivo is due to editing of the noncognate amino acid. PMID- 8422965 TI - In vitro dimerization of HIV-2 leader RNA in the absence of PuGGAPuA motifs. AB - Retroviral particles contain a dimeric genome consisting of two full-length, noncovalently linked RNA molecules. Linkage of the two genomes is thought to be critical for a productive reverse transcription reaction and may increase genetic recombination rates. The molecular nature of the dimer linkage structure (DLS) is poorly understood. It was recently shown that in vitro synthesized retroviral transcripts can dimerize in the absence of protein factors. We studied in vitro dimerization of human immunodeficiency virus type 2 (HIV-2) RNA. Specific dimerization of HIV-2 RNAs was observed upon incubation at 37 degrees C in high salt buffer. Previously, physical and biochemical studies have mapped dimer linkage structures in retroviral leader RNA close to the gag open reading frame. In this study, we found efficient dimerization of HIV-2 RNAs containing only the 5' terminal 255 nucleotides of the leader RNA. Therefore, it seems likely that multiple dimerization signals are present in retroviral leader RNA. The implications for genome dimerization and genome packaging are discussed. PMID- 8422966 TI - 5S rRNA modification in the hyperthermophilic archaea Sulfolobus solfataricus and Pyrodictium occultum. AB - The 5S rRNAs from Sulfolobus solfataricus and Pyrodictium occultum were digested to nucleosides and analyzed using directly-combined HPLC/mass spectrometry. P. occultum 5S rRNA contains two modified nucleoside species, N4-acetylcytidine (ac4C) and N4-acetyl-2'-O-methylcytidine (ac4Cm). Oligonucleotides were generated from P. occultum 5S rRNA by RNase T1 hydrolysis, and their molecular weights were determined using electrospray mass spectrometry and compared with those predicted from the P. occultum 5S RNA gene sequence. Deviation in mass between expected and observed molecular weights permitted ac4Cm to be located at position 35, in the nonanucleotide CAA-CACC[ac4Cm]G, and the ac4C in one or both of two (C,U)G trinucleotides. 2'-O-Methylcytidine is unambiguously characterized in S. solfataricus 5S rRNA, confirming earlier tentative assignments at the analogous sequence position (Stahl, D.A., Luehrsen, K.R., Woese, C.R., and Pace, N.R. (1981) Nucleic Acids Res., Vol. 9, pp. 6129-6137; Dams, E., Londei, P., Cammarano, P., Vandenberghe, A., and De Wachter, R. (1983) Nucleic Acids Res. Vol. 11, pp. 4667-4676). Potential effects of the presence of ac4C and ac4Cm on thermal stabilization of 5S rRNA in thermophiles are discussed. PMID- 8422967 TI - HIV-1 TAR RNA-binding proteins control TAT activation of translation in Xenopus oocytes. AB - Human immunodeficiency virus (HIV-1) gene expression is activated by the viral TAT protein that interacts with an RNA sequence, TAR, located at the 5' end of all viral mRNAs. TAT functions primarily as a transcriptional activator in mammalian cells. However, in Xenopus oocytes TAT functions primarily as a translational activator. TAR is an RNA structure comprising a partially base paired stem, a tripyrimidine bulge in the upper stem, and an unpaired six nucleotide loop. In vitro, TAT binds directly to the bulge with no requirement for the loop. In vivo, however, mutations in the loop abolish TAT activation of transcription and translation, implying a requirement for TAR-binding cellular factors. We now provide genetic evidence for the presence of two TAR-specific cellular factors in Xenopus oocytes. These factors display independent and mutually exclusive interactions with either the loop or the bulge region of TAR. Furthermore, by using in vivo RNA competition assays we show that the cellular factors regulate the accessibility of the TAT binding site. The fact that Xenopus oocytes contain factors that specifically interact with a human viral RNA sequence might indicate that the TAT/TAR interaction is subverting a conserved pathway in the cell. PMID- 8422968 TI - Unique phylogenetic position of Diplomonadida based on the complete small subunit ribosomal RNA sequence of Giardia ardeae, G. muris, G. duodenalis and Hexamita sp. AB - Complete small-subunit rRNA (SSU-rRNA) coding region sequences were determined for two species of the intestinal parasite Giardia: G. ardeae and G. muris, both belonging to the order Diplomonadida, and a free-living member of this order, Hexamita sp. These sequences were compared to published SSU-rDNA sequences from a third member of the genus Giardia, G. duodenalis (often called G. intestinalis or G. lamblia) and various representative organisms from other taxa. Of the three Giardia sequences analyzed, the SSU-rRNA from G. muris is the smallest (1432 bases as compared to 1435 and 1453 for G. ardeae and G. duodenalis, respectively) and has the lowest G+C content (58.9%). The Hexamita SSU-rRNA is the largest in this group, containing 1550 bases. Because the sizes of the SSU-rRNA are prokaryotic rather than typically eukaryotic, the secondary structures of the SSU rRNAs were constructed. These structures show a number of typically eukaryotic signature sequences. Sequence alignments based on constraints imposed by secondary structure were used for construction of a phylogenetic tree for these four taxa. The results show that of the four diplomonads represented, the Giardia species form a distinct group. The other diplomonad Hexamita and the microsporidium Vairimorpha necatrix appear to be distinct from Giardia. PMID- 8422969 TI - Bacterial diversity in a soil sample from a subtropical Australian environment as determined by 16S rDNA analysis. AB - In order to investigate the genetic diversity of streptomycetes in an acid forested soil sample from Mt. Coot-tha, Brisbane, Australia, cells were mechanically lysed within the soil matrix and genomic DNA was isolated and purified. 16S ribosomal (r)DNA was amplified by the polymerase chain reaction (PCR) method using one primer conserved for members of the domain Bacteria and a second designed specifically for streptomycetes and related taxa. PCR amplification products were cloned into phage vector M13 mp19 and the diversity of 16S rDNA genes was determined by sequence analysis and oligonucleotide probing of the resultant clone library. Comparison of partial 16S rDNA sequences with published sequences revealed that few sequences originated from streptomycetes. The majority of sequences belonged to members of the alpha subclass of Proteobacteria. Other clones were related to planctomycetes, actinomycetes, or represented novel lines of descent. Bacteria that are customarily isolated from soil of pH 4-7 such as thiobacilli, bacilli, spore- and nonsporeforming actinomycetes, and pseudomonads are represented in the clone library in small numbers or were not detected at all. Parameters influencing the recovery, amplification, quantification, and interpretation of genetic information from natural sites are discussed. PMID- 8422970 TI - Thrombospondins: structure and regulation of expression. PMID- 8422971 TI - Self-cleaving catalytic RNA. AB - We describe the structures and catalytic properties of several naturally occurring self-cleaving RNA motifs that give 2', 3' cyclic phosphate products. The hammerhead and hairpin motifs are derived from plant pathogenic RNAs and the delta motif is part of the human hepatitis delta element. A fourth motif from Neurospora is less well characterized. By assembling the self-cleaving RNAs from more than one oligoribonucleotide, the cleavage reaction can be examined under a variety of conditions and catalytic turnover can be demonstrated. Mutagenesis and chemical methods to introduce modified nucleotides allowed the structural requirements to be deduced. The role of divalent cations in the catalytic mechanism is discussed. PMID- 8422972 TI - How many catalytic RNAs? Ions and the Cheshire cat conjecture. AB - Three well-characterized RNA catalysts not only require Mg2+ for activity, but also bind a metal ion (or ions) within the active site, apparently in a catalytic rather than solely structural role. I suggest, in view of the general catalytic utility of bound ions, that catalytic RNAs be viewed as Cheshire cats, by dimming their complex three-dimensional ribonucleotide structure to leave only the sharp mineral parts in view. That is, catalytic RNAs may be viewed as metalloenzymes, with the burdens of catalysis frequently borne by specifically poised metal ions. Comparison to modern protein metalloenzymes predicts particular RNA metallocatalysts that may be possible presently, and in a hypothetical ancestral RNA world that did not encode peptide catalysts. In support of this view, known catalytic RNAs can be considered Cheshire catalysts; that is, they have apparent cognates among the protein metalloenzymes. PMID- 8422973 TI - Trans-splicing of pre-mRNA in plants, animals, and protists. AB - Messenger RNA maturation in eukaryotes typically involves the removal of introns from long precursor molecules. An unusual form of RNA splicing in which separate precursor transcripts contribute sequences to the mature mRNA through intermolecular reactions has now been documented in a number of diverse organisms. In this review, the phenomenon of pre-mRNA trans-splicing has been divided into two categories. The "spliced leader" type, found in protozoans such as trypanosomes and lower invertebrates such as nematodes, results in the addition of a short, capped 5' noncoding sequence to the mRNA. The "discontinuous group II intron" form of trans-splicing, found in plant/algal chloroplasts and plant mitochondria, involves the joining of independently transcribed coding sequences, presumably through interactions between "intronic" RNA pieces. Both categories of trans-splicing are mechanistically similar to conventional nuclear pre-mRNA cis-splicing; potential evolutionary relationships are discussed. PMID- 8422974 TI - Small nuclear RNAs in messenger RNA and ribosomal RNA processing. AB - Ribonucleoproteins (RNPs) play essential roles in many aspects of gene expression. Two families of nuclear RNPs are involved in the processing of primary transcripts made by RNA polymerases I and II (pol I and II), two of the three polymerases present in the nuclei of eukaryotic cells. Ribosomal RNA precursor transcription by pol I, subsequent processing of the precursor, and the initial steps of ribosome assembly all take place in the nucleolus. A group of nucleolar RNPs containing small RNAs (small nucleolar RNAs or snoRNAs) are involved in the posttranscriptional nucleolar events of ribosome biosynthesis. Six members of a related family of small nuclear RNAs (snRNAs) are required for the processing of mRNA precursors in the nucleoplasm. Five of these snRNAs (U1, U2, U4-6) participate in the removal of intervening sequences while the sixth (U7) plays an essential role in the 3' processing of a subset of mRNA precursors, the histone pre-mRNAs. This is a review of structural and functional aspects of the U1-U7 snRNAs and of snoRNAs. PMID- 8422975 TI - RNA editing in kinetoplastid mitochondria. AB - RNA editing in the mitochondrion of kinetoplastid protozoa results in the posttranscriptional addition and deletion of uridine residues in mRNAs. Editing of mRNAs can lead to the formation of initiation codons for mitochondrial translation, the correction of frame-shifted genes at the RNA level, and in extensively edited mRNAs, the formation of complete reading frames. Kinetoplastid RNA editing requires that genetic information from two or more separately transcribed genes be brought together to form the mature, edited mRNA. The information necessary for the proper insertion or deletion of uridines in the mRNA is present in small mitochondrial transcripts termed guide RNAs (gRNAs). Editing of mRNAs appears to be associated with a high molecular weight complex, called the editosome, containing specific gRNAs, unedited mRNAs, and proteins. Editing is likely a two-step process involving first the breakage of a phosphodiester bond at the editing site and formation of a chimeric molecule with a gRNA covalently joined to the 5' end of the 3' portion of an mRNA. The chimera is resolved by the rejoining of the 5' end of the mRNA to the 3' portion of the mRNA with the addition or deletion of a uridine at the junction point. Two models are proposed for the biochemical mechanism of RNA editing. The first is an enzymatic cascade of cleavage and ligation while the other supports successive rounds of transesterification. The obvious functional necessity for editing in kinetoplastid mitochondria is the formation of translatable mRNAs. Far less clear is the evolutionary origin of editing and the role editing plays in regulating mitochondrial gene expression. PMID- 8422976 TI - RNA editing in plant mitochondria and chloroplasts. AB - In the mitochondria and chloroplasts of flowering plants (angiosperms), transcripts of protein-coding genes are altered after synthesis so that their final primary nucleotide sequence differs from that of the corresponding DNA sequence. This posttranscriptional mRNA editing consists almost exclusively of C to-U substitutions. Editing occurs predominantly within coding regions, mostly at isolated C residues, and usually at first or second positions of codons, thereby almost always changing the amino acid from that specified by the unedited codon. Editing may also create initiation and termination codons. The net effect of C-to U RNA editing in plants is to make proteins encoded by plant organelles more similar in sequence to their nonplant homologs. In a few cases, a strong argument can be made that specific C-to-U editing events are essential for the production of functional plant mitochondrial proteins. Although the phenomenon of RNA editing in plants is now well documented, fundamental questions remain to be answered: What determines the specificity of editing? What is the biochemical mechanism (deamination, base exchange, or nucleotide replacement)? How did the system evolve? RNA editing in plants, as in other organisms, challenges our traditional notions of genetic information transfer. PMID- 8422977 TI - Transfer RNA identity. AB - Since its theoretical inception, the tRNA molecule has been identified and many of its functions have been described in terms of its structure. However, the molecular basis for the aminoacylation specificity of tRNA has remained largely unsolved until recently, when substantial progress has disclosed fundamental aspects of this process. What we know and would like to know about tRNA identity will be outlined, as will the experimental approaches used in this research. PMID- 8422978 TI - Recognition of tRNAs by aminoacyl-tRNA synthetases. AB - Our present understanding of the molecular mechanisms responsible for the recognition of tRNAs by their cognate aminoacyl-tRNA synthetases (aaRS) is essentially based on three sources of information: 1) the characterization of tRNA identity determinants using in vivo and in vitro approaches, 2) the classification of synthetases from primary sequence analysis: aaRS can be partitioned into two classes according to the spatial structure of their ATP binding domain, and 3) the structural results of crystallographic investigations and solution studies. The crystal structures of three aaRS and two complexes, one of each class, are known to atomic resolution. tRNA recognition has two structural components. The interaction between the acceptor end and the active site domain is class-specific and the binding mode of the stem observed in the crystal structures of GlnRS-tRNA(Gln) and AspRS-tRNA(Asp) complexes can be generalized to their respective classes. Identity determinants located in other parts of the tRNA molecule are decoded by different domains of the enzyme. These protein modules exhibit a large structural diversity. The recognition process is then system or subgroup specific. PMID- 8422979 TI - tRNA-rRNA interactions and peptidyl transferase. AB - The extent to which ribosomal RNA is directly involved in the function of ribosomes has important implications for both the mechanism of translation and the molecular origins of life. Detailed evidence has accumulated that places the anticodon and acceptor ends of tRNA in close proximity to conserved features of rRNA in the ribosome. Recent studies are providing evidence that these features are important for ribosomal function. PMID- 8422980 TI - The frog prince-ss: a molecular formula for dorsoventral patterning in Xenopus. PMID- 8422981 TI - Each hypersensitive site of the human beta-globin locus control region confers a different developmental pattern of expression on the globin genes. AB - We have tested the effect of the individual DNase I hypersensitive site (HS) regions of the globin locus control region (LCR) on the developmental expression pattern of the human gamma and beta genes in transgenic mice. The results show that HS3 is the most active site during the embryonic period. It is also the only site capable of high level expression of the gamma genes during fetal hematopoiesis, in a population of cells that are capable of expressing both the gamma and beta genes. Region HS4 shows the highest activity during the adult stage and expresses the gamma genes only at low levels during the embryonic period. HS2 drives equivalent levels of gamma or beta transgene expression throughout development. HS1 has a similar pattern to HS2, although the activity of HS1 is very low. From these results we conclude that the HS regions have distinct developmental specificities and suggest that in the complete LCR they interact with each other to form a larger complex which, in turn, interacts with the globin genes. PMID- 8422982 TI - Interactions between Xwnt-8 and Spemann organizer signaling pathways generate dorsoventral pattern in the embryonic mesoderm of Xenopus. AB - This study analyzes the hierarchy of signals that spatially restrict expression of Xenopus Xwnt-8 to mesodermal cells outside of the Spemann organizer field and examines the potential role that endogenous Xwnt-8 may play in dorsoventral patterning of the embryonic mesoderm. The effects of ectopic introduction of a Nieuwkoop center-like activity or of ectopic expression of goosecoid, on the distribution of endogenous Xwnt-8 transcripts were analyzed. The results of these studies are consistent with the hypothesis that maternally derived signals from the Nieuwkoop center function to positively regulate expression of the homeo box gene goosecoid in Spemann organizer cells, leading to a subsequent repression of Xwnt-8 expression in these cells. This exclusion of Xwnt-8 from cells of the organizer field may be important for normal dorsal development, in that ectopic expression of Xwnt-8 in organizer cells after the midblastula stage, by injection of plasmid DNA, ventralizes the fate of these cells. This is distinct from the previously observed dorsalizing effect of Xwnt-8 when expressed prior to the midblastula stage by injection of RNA. The effects of plasmid-derived Xwnt-8 on isolated blastula animal cap ectoderm were also analyzed. Expression of Xwnt-8 in animal pole ectoderm after the midblastula stage ventralizes the response of dorsal animal pole cells to activin and allows naive ectodermal cells to differentiate as ventral mesoderm in the absence of added growth factors. Collectively, these data are consistent with the hypothesis that Xwnt-8 plays a role in the mesodermal differentiation of ventral marginal zone cells during normal development. Furthermore, endogenous Xwnt-8 may ventralize the response of lateral mesodermal cells to dorsalizing signals from the organizer, thus contributing to the graded nature of the final body pattern. PMID- 8422983 TI - SinI modulates the activity of SinR, a developmental switch protein of Bacillus subtilis, by protein-protein interaction. AB - SinR, a 111-amino-acid DNA-binding protein, is a pleiotropic regulator of several late growth processes in Bacillus subtilis. It acts as a developmental switch, positively regulating genes for competence and motility and repressing aprE and stage II sporulation genes. It is encoded by the second gene in a two gene operon, but previous results have also indicated that these two genes are differently regulated. We show in this discussion that the product of sinI, the first open reading frame (ORF) of this operon, interferes with the function of SinR. In vivo experiments have demonstrated that overexpression of sinI results in phenotypes that are observed in cells with a null mutation of sinR. A chromosomal in-frame deletion of sinI gives rise to a phenotype associated with higher levels of SinR. Thus, SinI acts as an antagonist to SinR. In vitro experiments have shown that the interaction between these two proteins is a direct one. SinI prevents SinR from binding to its target sequence on aprE, and the two proteins form a complex that can be immunoprecipitated with antibodies to either SinR or SinI. PMID- 8422984 TI - Role of the sigma 70 subunit of RNA polymerase in transcriptional activation by activator protein PhoB in Escherichia coli. AB - Transcription of the genes belonging to the phosphate (pho) regulon in Escherichia coli, which are induced by phosphate starvation, requires the specific activator protein PhoB in addition to the RNA polymerase holoenzyme containing the major sigma-factor sigma 70. To study the mechanism of transcriptional activation and identify the subunit of RNA polymerase involved in specific interaction with PhoB, we attempted to isolate rpoA and rpoD mutants that are specifically defective in the expression of the pho genes. We isolated two rpoD mutants with such properties, but no rpoA mutant with similar properties. The rpoD mutations altered amino acids within and near the first helix of the putative helix-turn-helix (HTH) motif in the carboxy-terminal region of sigma 70. Activities of the pho promoters in vivo were severely reduced in these mutants, whereas those of the PhoB-independent promoters were affected only marginally at most. The reconstituted mutant RNA polymerase holoenzymes were severely defective in transcribing the pstS gene, one of the pho genes, whereas they were efficient in transcribing the PhoB-independent promoters. Phosphorylated PhoB, which binds to the pho promoters with high affinity, mediated the specific binding of the wild-type holoenzyme to the pstS promoter, but it did not mediate the binding of the mutant holoenzymes. These results suggest that PhoB promotes specific interaction between RNA polymerase and the pho promoters for transcriptional activation, and the first helix of the putative HTH motif plays an essential role in the interaction, probably by making direct contact with PhoB. PMID- 8422985 TI - Elongation factor NusG interacts with termination factor rho to regulate termination and antitermination of transcription. AB - NusG is a transcriptional elongation factor in Escherichia coli that aids transcriptional antitermination by the phage lambda N protein. By using NusG affinity chromatography, we found that NusG binds directly and selectively to termination factor rho. NusG was shown previously to be needed for termination by rho in vivo, and we show here that NusG increases the efficiency of termination by rho at promoter-proximal sites in vitro. The rho026 mutation makes termination by rho less dependent on NusG. It also makes antitermination by N at rho dependent terminators and the binding of rho to NusG temperature sensitive. Therefore, the interaction of NusG with rho is important both for rho-dependent termination and for antitermination by N at rho-dependent terminators. PMID- 8422986 TI - Morphogenesis in Drosophila requires nonmuscle myosin heavy chain function. AB - We provide the first link between a known molecular motor and morphogenesis, the fundamental process of cell shape changes and movements that characterizes development throughout phylogeny. By reverse genetics, we generate mutations in the Drosophila conventional nonmuscle myosin (myosin II) heavy chain gene and show that this gene is essential. We demonstrate that these mutations are allelic to previously identified, recessive, embryonic-lethal zipper mutations and thereby identify nonmuscle myosin heavy chain as the zipper gene product. Embryos that lack functional myosin display defects in dorsal closure, head involution, and axon patterning. Analysis of cell morphology and myosin localization during dorsal closure in wild-type and homozygous mutant embryos demonstrates a key role for myosin in the maintenance of cell shape and suggests a model for the involvement of myosin in cell sheet movement during development. Our experiments, in conjunction with the observation that cytokinesis also requires myosin, suggest that the processes of cell shape change in morphogenesis and cell division are intimately and mechanistically related. PMID- 8422987 TI - Sex-specific transcriptional regulation by the male and female doublesex proteins of Drosophila. AB - The somatic sexual phenotype of Drosophila is regulated by the sexual differentiation pathway. Male (DSXM) and female (DSXF) proteins encoded by doublesex (dsx), a gene at the end of this pathway, bind to three sites within a 127-bp enhancer that directs sex- and tissue-specific transcription of Yolk protein genes. We describe mutagenesis of these binding sites and the resulting effects on DSXM and DSXF binding in vitro and on gene regulation in wild-type and dsx mutant flies. The results demonstrate that DSXM represses and DSXF activates transcription from the two strongest binding sites. Thus, the pathway regulates sex-specific transcription through the male and female dsx proteins that act directly on the target gene, but with opposite effects. PMID- 8422988 TI - Tissue-specific RNA splicing generates an ankyrin-like domain that affects the dimerization and DNA-binding properties of a bHLH protein. AB - mRNAs encoding two rat bHLH proteins, referred to as REB alpha and REB beta, have been identified as alternatively spliced transcripts derived from a single genomic locus. Alternative RNA processing events results in tissue-specific differences in the ratios of these two mRNAs. Although it exhibits a highly enriched level of expression in the developing neural tube, the REB gene is expressed at variable levels in many organs of the mature animal. The REB alpha sequence contains a region characterized by a leucine heptad repeat that is situated amino-terminal of the carboxy-terminally located bHLH domain. REB beta is identical to REB alpha except for a 24-amino-acid insertion in the leucine heptad repeat that results from the inclusion of an additional 72-bp exon in the REB beta transcript. As a consequence of this insertion, REB beta exhibits a markedly diminished capacity to bind to cognate E-box-binding sites and to form homodimers and heterodimers with other members of the bHLH gene family. Analysis of the 24-amino-acid REB beta-specific insert revealed that it mediates an inhibitory function and exhibits a significant degree of sequence similarity to ankyrin-like repeats. It is proposed that this tissue-specific pattern of REB RNA splicing is involved in the determination of corresponding tissue-specific combinations of heterodimeric complexes of ubiquitous and tissue-restricted bHLH proteins. Thus, REB alpha and REB beta represent a novel example of a regulated formation of an ankyrin-like domain within a bHLH protein, thereby mediating control of protein-protein interactions. PMID- 8422989 TI - Differential positive control by Oct-1 and Oct-2: activation of a transcriptionally silent motif through Oct-1 and VP16 corecruitment. AB - Transcriptional regulation by the ubiquitous human POU homeo domain protein Oct-1 and the related B-cell protein Oct-2 is a model for understanding how proteins that recognize the same regulatory site elicit different programs of gene transcription. Here, we describe a mechanism for differential promoter activation whereby only Oct-1, through selective corecruitment with the herpesvirus trans activator VP16, acquires the ability to stimulate transcription from a TAATGARAT containing site that responds to neither Oct-1 nor Oct-2 alone. To measure differential in vivo activation by human Oct-1 and Oct-2 in response to VP16, we have developed a transient assay in murine NIH-3T3 cells. Surprisingly, murine Oct-1 associates with VP16 much less effectively than its human counterpart, most likely because the murine Oct-1 homeo domain differs at four positions from the human Oct-1 homeo domain. The murine cell transient assay shows directly that human Oct-1, but not human Oct-2, can respond to VP16 in vivo. The Oct-1 DNA binding POU domain is sufficient and the Oct-1 homeo domain is critical for this response, because an Oct-1 POU domain containing the Oct-2 homeo domain fails to respond to the VP16-induced positive control of transcription. Thus, by selective homeo domain interaction and corecruitment to an otherwise silent regulatory element, VP16 expands the repertoire of sites responsive to Oct-1 without affecting the activity of its close relative Oct-2. PMID- 8422990 TI - Circular forms of developmentally excised DNA in Euplotes crassus have a heteroduplex junction. AB - Extensive DNA elimination via a DNA breakage and rejoining process occurs during macronuclear development in the hypotrich ciliate Euplotes crassus. The excision process involves the removal of short, unique segments of DNA (internal eliminated sequences; IESs) and at least two highly repetitive families of transposon-like elements (Tec elements). Previous studies have demonstrated that circular forms of both IESs and Tec elements are generated following their developmental excision and that some flanking DNA sequences are retained at the circle junctions. In this study we have further analyzed the circle junctions of IESs. Analysis of polymerase chain reaction (PCR) products derived from IES circle junctions indicates that at least two sequence arrangements can be present. The circle junctions contain both of the direct repeats that define the ends of the IES separated by either 2 bp flanking the right end of the IES and 8 bp from the left-flanking region, or 8 bp from the right and 2 bp from the left. Using a method that we have termed "strand-biased PCR," we obtained evidence that the junctions of free circular IESs have a 6-base heteroduplex at their center, such that one strand of the DNA is derived from the left-flanking region of the IES and the other from the right. Models of IES excision are presented that incorporate these results and those of previous studies on the excision process. PMID- 8422991 TI - Increased antibody expression from Escherichia coli through wobble-base library mutagenesis by enzymatic inverse PCR. AB - We tested the value of a new library mutagenesis approach, called library enzymatic inverse PCR (LEIPCR), for expression-level enhancement of antibody Fv fragments produced in Escherichia coli. The production level of active, metal chelate-specific antibody from our constructs is limited by a low expression level of the second, heavy-chain cistron. To increase the production level, LEIPCR was applied to the wobble bases of the second cistron leader peptide. In LEIPCR mutagenesis, the entire plasmid is amplified using mutagenic primers with class-IIS restriction endonuclease (ENase) sites at their 5' ends. The PCR product is digested with the class-IIS ENase (here, BsaI; GGTCTCN[symbol: see text]NNNN[symbol: see text]), which removes its own recognition sequence, and the ends are self-ligated. Thus, LEIPCR can be used to make plasmid mutant libraries regardless of the nucleotide sequence, and independent of available ENase sites. The resulting library of 10(7) wobble mutants was screened for active Fv by a colony filter lift. A selected mutant was shown to produce between four- and elevenfold more active Fv than the wild type (wt), and fivefold more heavy chain. Mutations outside of the leader peptide were shown not to be involved. The mutated areas of the mRNAs of two different up-mutants may have less secondary structure than the wt. Thus, the sequence of the mRNA of the second leader peptide was limiting to the expression level of heavy-chain and active Fv. PMID- 8422992 TI - Endoglucanase CasA from alkalophilic Streptomyces strain KSM-9 is a typical member of family B of beta-1,4-glucanases. AB - CasA is an endo-beta-1,4-glucanase from Streptomyces KSM-9 belonging to family B of beta-1,4-glucanases. A previous analysis of a portion of the corresponding gene (casA) revealed sequencing errors in a region encoding part of the catalytic site. Additional errors in the original sequence were suspected, based on sequence comparison of the C terminus of CasA with other members of its family. Re-sequencing of the remainder of the casA coding region showed that CasA is a typical member of family B. PMID- 8422993 TI - Cloning, characterization, and expression in Streptomyces lividans 66 of an extracellular lipase-encoding gene from Streptomyces sp. M11. AB - A gene encoding an extracellular lipase from Streptomyces sp. M11 was cloned in the high-copy-number vector pIJ486, using S. lividans 66 as host. A 28-kDa protein was secreted by S. lividans carrying pB13, which harbors a 6-kb insert, and identified as the product of the cloned gene. Comparison of the N-terminal amino acid (aa) sequence of the purified extracellular lipase with the nucleotide (nt) sequence of the lip gene revealed the presence of a 48 aa long signal peptide. The nucleotide sequence also revealed the presence of a motif, Gly-His Ser-Met-Gly, similar to the one found surrounding the active-site Ser in other lipases. The gene is most likely monocistronic. Subcloning experiments indicated that another gene might be required for high-level expression, since subcloning of the structural gene alone resulted in diminished extracellular lipase activity. The lipase gene promoter was identified by S1 mapping experiments, and found to be similar to other Streptomyces vegetative promoters. PMID- 8422994 TI - Cloning and characterisation of an aminopeptidase P-encoding gene from Streptomyces lividans. AB - An aminopeptidase P (PepP)-encoding gene has been cloned from Streptomyces lividans 66 by screening for overexpression of activity using the chromogenic substrate Gly-Pro-beta-naphthylamide as a liquid overlayer on colonies growing on agar medium. The pepP gene was localised by deletion mapping, and the nucleotide sequence was determined. The deduced amino acid sequence was found to display significant similarity to Escherichia coli PepP. The partially purified S. lividans enzyme had a 50-kDa subunit and was present as a homodimer. Direct Edman degradation of the purified protein confirmed that pepP encoded the observed intracellular PepP. PMID- 8422995 TI - Characterization of the gene coding for the Rickettsia prowazekii DNA primase analogue. AB - The gene (dnaG) coding for DNA primase in the obligate intracellular parasitic bacterium, Rickettsia prowazekii, has been isolated and characterized. An open reading frame (ORF) of 1848 bp capable of encoding 616 amino acids (aa) is located 18 bp upstream from the gene coding for the major sigma factor of R. prowazekii, sigma 73. Based on aa sequence comparisons of DNA primase from R. prowazekii, Escherichia coli, Salmonella typhimurium and Bacillus subtilis, we propose that R. prowazekii dnaG begins 69 bp into the ORF and encodes 593 aa with a calculated M(r) of 68,683. An upstream ORF overlaps 66 of the first 69 bp of the larger R. prowazekii dnaG ORF, suggesting either an overlapping gene structure or the generation of the smaller protein product of 593 aa. Predicted aa sequence of R. prowazekii primase compared to E. coli, S. typhimurium and B. subtilis primases reveals 30.5%, 30.5% and 29.7% aa identity, respectively. The R. prowazekii dnaG gene failed to complement an E. coli dnaG temperature sensitive mutation perhaps due to poor expression of the gene or inability to function properly in E. coli. The gene organization of an ORF followed by DNA primase (dnaG) and then the major sigma factor (rpoD) is consistent with the major macromolecular synthesis operons of E. coli, S. typhimurium and B. subtilis. PMID- 8422996 TI - Thiamine-repressible expression vectors pREP and pRIP for fission yeast. AB - The promoter and polyadenylation signal of the thiamine-repressible gene nmt1 of Schizosaccharomyces pombe have been used to construct the pREP extrachromosomally replicating plasmids and the pRIP integrative expression plasmids. These plasmids permit thiamine-mediated control of transcription to be applied to cloned genes. PMID- 8422997 TI - TATA box mutations in the Schizosaccharomyces pombe nmt1 promoter affect transcription efficiency but not the transcription start point or thiamine repressibility. AB - The nmt1 gene of Schizosaccharomyces pombe is highly expressed and subject to transcriptional repression by thiamine. The nmt1 promoter, in common with other strong promoters in this organism, contains a canonical sequence element, 5' ATATATAAA, located 25 bp upstream from the transcription start point (tsp). We have made stepwise deletions of the TATA box and quantitated the effects of the mutations by assaying the expression of the chloramphenicol acetyltransferase (CAT)-encoding gene (cat) cloned downstream. Our results demonstrate that progressive truncation of the TATA box results in a concomitant decrease in promoter strength as judged both by the loss of CAT activity in cell extracts and by a reduction in the steady-state level of cat mRNA. Both the induced level and the residual, repressed level of expression observed in the presence of thiamine are similarly down-regulated. On the other hand, even in the most extreme mutant, the tsp is unaffected, suggesting that other elements in the nmt1 promoter are important in determination of the tsp. The properties of the modified promoters have made them useful for extending the range of the pREP inducible expression vectors. PMID- 8422999 TI - Complete sequence of the Salmonella typhimurium gene encoding malate dehydrogenase. AB - The nucleotide (nt) sequence of mdh (encoding malate dehydrogenase) in Salmonella typhimurium is presented. The relative positions of argR (encoding arginine repressor) and mdh on the chromosome of S. typhimurium were determined from analysis of the nt sequence. The homology of the 3'-terminal end with only one of two published mdh sequences in Escherichia coli identifies the correct sequence in this organism. PMID- 8422998 TI - Cloning and characterization of an alpha-tubulin-encoding gene from rat-derived Pneumocystis carinii. AB - The gene and cDNA (TUB1) encoding an alpha-tubulin (alpha-Tub) from Pneumocystis carinii carried in rats were isolated and sequenced. The gene produced a 1.4-kb transcript that contained an open reading frame of 1347 nucleotides coding for a protein of 50,326 Da. The nt sequence of TUB1 of P. carinii was 65-71% identical to other TUB1 genes, including those of yeast and human. The deduced amino acid sequence of P. carinii alpha-Tub showed 75-84% identity to other alpha-Tubs. Eight small introns resided within TUB1. P. carinii TUB1 was mapped to a 425-kb chromosome. The cloned gene appears to be the only alpha-Tub-encoding gene in the P. carinii genome. PMID- 8423000 TI - The nifH gene encoding the Fe protein component of the molybdenum nitrogenase from Azotobacter chroococcum. AB - The nucleotide sequence spanning the nifH gene and part of the nifD gene encoding the molybdenum nitrogenase from Azotobacter chroococcum was determined. The transcription start point of the nifH promoter was mapped, and a potential transcriptional attenuator was located between the nifH and nifD genes. PMID- 8423001 TI - Nucleotide sequences of the trpI, trpB, and trpA genes of Pseudomonas syringae: positive control unique to fluorescent pseudomonads. AB - A 904-bp probe from Pseudomonas aeruginosa was used to identify the trpB, trpA and trpI genes of Pseudomonas syringae. Transcription initiation at the P. syringae trpBA promoter in vitro was activated by the P. aeruginosa TrpI protein in the presence of indoleglycerol phosphate. Thus, trpB and trpA are regulated positively in three species of fluorescent pseudomonads, P. aeruginosa, P. putida, and P. syringae, but in no other eubacteria so far investigated [Crawford, Annu. Rev. Microbiol. 43 (1989) 567-600]. In addition to conservation of protein-coding sequences, there is a high degree of nucleotide sequence identity in the intergenic control region that includes the divergent trpI and trpBA promoters, especially in the binding sites for TrpI protein. Differences in patterns of codon usage distinguish the trpI genes of P. syringae and P. putida from P. aeruginosa trpI and from the trpB and trpA genes of all three species. PMID- 8423002 TI - Application of the mini-mu phage for the isolation of lac transcriptional fusions in Bacillus subtilis genes. AB - A cassette containing a selectable cat gene and the lacZ gene without its own promoter has been incorporated into the mini-Mu bacteriophage genome. This mini Mu derivative, referred to as mMu-Bs, can be used in Escherichia coli for the generation of lacZ transcriptional fusions to Bacillus subtilis genes cloned into plasmids. The resultant fusions can be analyzed in B. subtilis either as multicopy plasmids or as a single copy integrated via a Campbell-like recombination into the wild-type locus of the cloned fragment. PMID- 8423003 TI - Control of gonococcal pilin-encoding gene expression in Escherichia coli. AB - The pilE gene from Neisseria gonorrhoeae, unlike other type-4 pilin-encoding genes, is well expressed in Escherichia coli. Two putative promoters have been implicated in the transcription of this gene. Besides the -24/-12 promoter used to transcribe type-4 pilin-encoding genes in most species, the consensus sequence for a conventional promoter is also present. The two promoters overlap and would have almost identical transcription start points (tsp). Transcription from a -24/ 12 promoter should be abolished in an E. coli rpoN mutant. A recombinant plasmid carrying pilE could not be transformed into such a mutant, apparently because the synthesis of the N-terminal hydrophobic domain of pilin is lethal to the rpoN mutant. This suggests that pilE is expressed at a higher level in an rpoN mutant than it is in a wild-type (wt) strain of E. coli. This suggestion was confirmed by constructing fusions between the pilE promoter region and a promoter-less cat gene. We suggest that the conventional promoter is primarily responsible for the transcription of pilE, but that the binding of the RpoN sigma factor partially represses transcription of this gene in wt strains. In an rpoN mutant, the repression is removed and transcription occurs at a level that is lethal to the mutant host. PMID- 8423004 TI - Analysis of hemolysin operons in Actinobacillus pleuropneumoniae. AB - Among the twelve different serotypes of Actinobacillus pleuropneumoniae, the causative agent of swine pleuropneumonia, a strongly active hemolysin I (HlyI) is produced by serotypes which are particularly virulent, and less active hemolysin II (HlyII) is produced by all serotypes except type 10. In the serotypes 1, 5a, 5b, 9, 10 and 11, which produce HlyI, the hemolysin (hly) operon consists of a structural hlyIA gene, encoding pre-HlyI, an activator gene, hlyIC, necessary for the activation of pre-Hly to active Hly, and two genes, hlyIB and hlyID, involved in Hly secretion. These genes are clustered in the order, hlyICABD. This is characteristic to RTX toxin (repeats in the structural toxin) operons. The HlyII operons in all serotypes producing HlyII consist only of the pre-HlyII-encoding gene, appA, and its activator gene, appC. The serotypes, which produce HlyII, but not HlyI, contain a truncated HlyI operon, with the promoter, hlyIB and hlyID, and a small segment of the C terminus of hlyIA. This partial HlyI operon might have been formed by deletion of hlyIC and most of hlyIA. In serotype 3, which produces HlyII, but no HlyI, and which releases only minute amounts of this Hly into the growth medium, none of the hlyI genes and consequently no Hly secretion genes were found. The above results postulate that HlyII is secreted via the products of hlyIB and hlyID, and explain the low amount of HlyII secreted by serotype 3. Cloning and analysis of the structural genes encoding pre-HlyI and pre-HlyII among the different serotypes revealed differences in the hlyIA genes which are highly similar in the serologically related serotypes 1, 9 and 11, and differ from the serotypes, 5a, 5b and 10. The hlyIIA genes, in contrast, seem to be conserved in all serotypes. PMID- 8423005 TI - A simplified method for generating step-wise deletions using PCR. AB - A simple and general method is described for the generation of ordered deletions for DNA sequencing. Nicked plasmids, the substrates for step-wise digestion by exonuclease III, are obtained after the self-ligation of PCR products with phosphorylated and non-phosphorylated primers and plasmid DNAs as template. The method is suitable for use with any plasmid vector and for generation of deletion clones with deletions in both possible directions. PMID- 8423006 TI - Deletion of the essential gene 24 from the bacteriophage T4 genome. AB - We deleted the essential gene 24 from the genome of bacteriophage T4. The delta 24 phage is a conditional lethal mutant that can grow only when the host strain supplies the product of gene 24 in trans, or when the phage acquires a functional gene 24 by some type of recombination event. Thus, gene 24 can be used as a selectable marker, for example permitting transposition into the T4 genome and analyses of plasmid-phage recombination [Woodworth and Kreuzer, Mol. Microbiol. 6 (1992) 1289-1296; H.W.E. and K.N.K., manuscript submitted]. We also found that the promoter region of gene 24 allows a low level of autonomous plasmid replication in T4-infected cells, raising the possibility of a previously unrecognized mode of T4 replication initiation. PMID- 8423007 TI - A protein required for secretion of cholera toxin through the outer membrane of Vibrio cholerae. AB - A gene essential for the secretion of cholera toxin from the periplasm of Vibrio cholerae into the extracellular medium has been isolated and its nucleotide sequence determined. It encodes a cytoplasmic protein of 56 kDa that exhibits a high degree of similarity to gene products required for extracellular protein secretion in several other Gram- organisms. Sequence similarities in its potential ATP-binding site suggest that the protein may act as an energy provider or signal transducer in the process of extracellular secretion. PMID- 8423008 TI - Cloning and characterization of a gene coding for the catechol 1,2-dioxygenase of Arthrobacter sp. mA3. AB - The catA gene, coding for the catechol 1,2-dioxygenase (C12O) of the bacterial strain Arthrobacter sp. mA3, was cloned and expressed in Escherichia coli. One plasmid containing a 6.1-kb EcoRI insert was selected by its ability to degrade catechol and to accumulate cis-cis-muconate. The DNA insert of this plasmid was mapped with restriction enzymes. The catA gene was subcloned on a 1.3-kb PstI EcoRI fragment by deleting the adjacent restriction fragments. The nucleotide sequence of catA was determined. The C12O is coded for by a gene spanning 849 nucleotides and the deduced M(r) of the protein is 30,560. The polypeptide encoded by the cloned catA gene was expressed in an E. coli minicell system and detected by gel electrophoresis. PMID- 8423009 TI - Inactivation of the Escherichia coli B41 (O101:K99/F41) rfb gene encoding an 80 kDa polypeptide results in the synthesis of an antigenically altered lipopolysaccharide in E. coli K-12. AB - The genetic organisation of the rfb region from Escherichia coli B41 (O101:K99/F41) which determines the biosynthesis of the O101 O-antigen of the lipopolysaccharide (LPS) has been examined in E. coli K-12. Maxicell analysis of the plasmid-encoded proteins facilitated the construction of a physical map of the rfb region, consisting of six proteins, designated A (87 kDa), B (80 kDa), C (49 kDa), D (38 kDa), E (36.5 kDa) and F (27 kDa). Proteins E and F are not required for O-antigen biosynthesis. The introduction of frameshift mutations within the region encoding protein B resulted in the synthesis of an antigenically altered LPS which is shorter than the wild-type LPS, as assessed by reaction to antisera in colony and Western immunoblots, and by silver staining of LPS separated on sodium dodecyl sulfate-polyacrylamide-gel electrophoresis. The results demonstrate that protein B has a novel role in O-antigen biosynthesis associated with both the control of LPS chain length and antigenic structure. The nucleotide sequence of the rfb gene encoding protein B has been determined, confirming it to be a 697-amino acid protein of 78.9 kDa predicted to be located in the cytoplasmic membrane. PMID- 8423010 TI - Sequence and arrangement of genes encoding enzymes of the acetone-production pathway of Clostridium acetobutylicum ATCC824. AB - The nucleotide sequence of three open reading frames in the acetone-production locus of Clostridium acetobutylicum ATCC824 has been established. The three gene products, corresponding to acetoacetate decarboxylase (EC 4.1.1.4) and both subunits of the acetoacetyl-CoA:acetate/butyrate:CoA transferase (EC 2.8.3.9) are transcribed in two convergently arranged operons. The intervening DNA region separating the two transcripts is characterized by an inverted repeat which appears capable of forming a stem-loop structure functioning as a Rho-independent transcription terminator in both directions. PMID- 8423011 TI - The use of degenerate, sensor gene-specific, oligodeoxyribonucleotide primers to amplify DNA fragments from Staphylococcus aureus. AB - The sensor proteins of bacterial two-component regulatory systems comprise a large family of proteins that are involved in environmental sensing and signal transduction. To study these proteins in the Gram+ pathogen, Staphylococcus aureus, two pairs of degenerate oligodeoxyribonucleotides (oligos) that corresponded to conserved sequences contained within sensor protein-encoding genes were synthesized. Using these oligo primers, DNA fragments from S. aureus were amplified by polymerase chain reaction (PCR), cloned in Escherichia coli, and sequenced. Comparison of the deduced amino acid sequences from these cloned fragments to the sequences contained in the GenBank database suggest that some of the PCR products were derived from sensor protein-encoding genes. However, several other fragments were identified that encoded peptides with up to 65% identity to transport proteins. Given the biochemical and functional properties of some of these proteins, these data suggest that sensor and transport proteins may be evolutionarily related. PMID- 8423012 TI - Is ovarian cancer a clonal disease? PMID- 8423013 TI - A portable tensiometer for assessing surgeon's knot tying technique. AB - The purpose of this study was to determine the influence of surgeon's tying technique on knot security using O and 2-O monofilament and multifilament nylon sutures. By use of an Instron Tensile Tester and a portable tensiometer, knot security was achieved with these sutures using four-throw square knots (1 = 1 = 1 = 1). After didactic and psychomotor skill training, medical students were taught to construct the four-throw square knot using either a two-hand tie or an instrument tie. By use of the portable tensiometer, their knot tying techniques were judged to be superior to those used by a cohort of obstetricians and gynecologists. The surgeon's faulty technique can easily be corrected by didactic information and psychomotor skill training. PMID- 8423014 TI - Management of advanced juvenile granulosa cell tumor of the ovary. AB - Juvenile granulosa cell tumor of the ovary occurs most frequently in young women and children. Ten percent of cases present during pregnancy. The majority of tumors are in FIGO Stage I and have a favorable prognosis. The prognosis of higher stage tumors, however, is generally less favorable. We report the long term, disease-free survival of a patient with FIGO Stage III juvenile granulosa cell tumor of the ovary. We believe this to be the first report of a successful pregnancy following "MAC" chemotherapy for this particular malignancy. PMID- 8423015 TI - Five primary malignancies in a single patient: a report of two cases and a review of the literature. AB - Two patients who developed five independent primary malignancies are reported. Each developed multiple distinct malignancies over 45 and 29 years, respectively. The medical literature and etiology of this rare phenomenon are discussed. PMID- 8423016 TI - Advanced ovarian carcinoma managed in an HIV-positive patient. AB - This is the first report of ovarian cancer developing in a patient with HIV infection. The patient had Stage IIIC, grade 2, serous ovarian carcinoma. Aggressive therapy consisting of primary cytoreduction, high-dose platinum-based chemotherapy, and second-look laparotomy was well tolerated. The patient's CA-125 level quickly normalized but microscopic disease was found at second-look laparotomy. Although ovarian carcinoma has not been reported in the HIV-positive population, the incidence is likely to increase as the HIV-positive patient population ages and the epidemic in women continues. Aggressive primary therapy can result in a significant response; however, further experience and follow-up will be necessary to determine if there is decreased survival in this patient group as has been reported with other HIV-associated malignancies. PMID- 8423017 TI - Radiosensitization of uterine cancer cell lines by cytotoxic agents. AB - Radiotherapy remains an integral part of uterine cancer therapy. Overcoming radioresistant tumors by sensitizers continues to be a prime objective in radiotherapy research. In this study, the effects of five cytotoxic agents on two radiosensitive and four radioresistant uterine cancer cell lines were investigated. The ATP bioluminescence was used to measure surviving fractions. Data analysis was done using the linear quadratic model and radiosensitivity index D. Both AN3 and SKUT1B were radiosensitive with Ds of 1.73 and 1.72 Gy, respectively. The resistant cell lines had the following D values: AE7, 3.50; ECC, 6.61; HEC1A, 4.59; and HEC1B, 13.49 Gy. The average radiosensitization effects for various drugs were measured by reduction of D: DXR 45 +/- 7, DDP 40 +/- 9, 5FU 55 +/- 10, MITO 59 +/- 14, and HU 1.7 +/- 7%. Except for HU, Wilcoxon analyses revealed that these sensitizing effects were significant with P < 0.02. In summary, Adriamycin, 5-fluorouracil, cisplatin, and mitomycin-C have the potential to be radiosensitizers in uterine cancer cell lines. PMID- 8423018 TI - Interstitial reirradiation for recurrent gynecologic malignancies: results and analysis of prognostic factors. AB - Thirteen patients with recurrent or new primary gynecologic malignancies after previous radiation therapy (RT) underwent interstitial reirradiation (IRI) from July 1986 through December 1990. Mean and median ages were 63 and 70 years, respectively. Mean and median implanted volumes were 14.3 and 12 cc, respectively. Overall, 9/13 (69%) had complete responses to IRI and 6 (46%) continue to have no evidence of disease (NED) 24-71 months later (median follow up, 59 months). Of 7 patients with recurrent cervical or new primary vaginal carcinoma, 5 (71%) remain free of disease 27-71 months (median, 58 months) after IRI. Of 6 patients with recurrent endometrial carcinomas, only 1 (16%) continues with NED 24 months after IRI. Patients with NED after IRI had a median disease free interval prior to IRI of 100 months compared to 6 months in patients failing IRI. Trends toward improved outcomes were observed in squamous vs adenocarcinoma, smaller tumor volumes, higher implant doses, and vaginal wall/suburethra vs vaginal cuff location. One possible complication, a rectovaginal fistula, developed in the presence of recurrent cervical cancer 22 months after IRI. Interstitial reirradiation is an effective treatment for selected patients with recurrent gynecologic malignancies after previous RT. Advantages of IRI over radical surgery include its potential to preserve organ structure and function and its applicability to patients with medical contraindications to salvage surgery. Furthermore, since subsequent exenterative surgery should not be compromised in patients failing IRI, a policy of IRI as initial treatment may be justified for patients in whom the potential for morbidity is limited. PMID- 8423019 TI - A new aggressive treatment approach to high-grade endometrial cancer of possible benefit to patients with stage I uterine papillary cancer. AB - Between January 1, 1988 and December 31, 1990, a new high-intensity treatment was delivered to 10 patients with clinical stage I uterine papillary serous carcinoma (UPSC) and 21 patients with clinical stage I, FIGO grade 3 endometrial adenocarcinoma. The treatment consisted of a radical hysterectomy and bilateral salpingo-oophorectomy with pelvic lymph node sampling, adjuvant external pelvic radiation, and four courses of cis-platinum/epirubicin. The clinical outcome was compared to that of a historical control group treated before 1988 with surgery and radiation. None of the high-intensity treated UPSC patients died or relapsed during a median observation time of 32 months. There was a significantly better survival in the high-intensity treated UPSC group compared to the controls. The UPSC patients diagnosed after January 1, 1988 showed a trend toward better survival compared to those diagnosed before this date, whether they had received high-intensity treatment or not. No significant difference in survival was observed in the non-UPSC group subjected or not to the high-intensity treatment. PMID- 8423020 TI - Treatment results and prognostic factors in a population-based study of epithelial ovarian cancer. AB - The Swedish Tumor Registries are population based. Three hundred eighty-four patients with epithelial ovarian cancer registered in the Tumor Registry of the Southeast Health Care Region during the years 1984 to 1987 were reviewed; 332 patients were eligible for survival analysis. The protocol treatment during this period included primary surgery aimed at tumor reduction followed by combination chemotherapy (anthracyclin + cis-platinum, AP) in FIGO stages IC-IV. All patients treated otherwise were registered as prescribed nonprotocol treatment. An overall corrected 5-year survival of 40% was recorded. However, only 166/325 of the patients (51%) were actually prescribed protocol treatment as defined above. The 5-year survival was 49% for patients prescribed protocol treatment compared to 33% for those prescribed nonprotocol treatment (P < 0.0001). Stages III-IV patients prescribed protocol treatment had 23% 5-year survival compared to 11% for patients prescribed nonprotocol treatment (P < 0.0001). The impact of cis platinum-based combination chemotherapy was thus not as pronounced as expected from clinical studies since many patients were not considered to be in a good enough general condition to receive cis-platinum. Multivariate Cox analysis of 223 cases showed that age (P < 0.0001), stage (P = 0.0002), grade of differentiation (P = 0.006), and postoperative residual tumor (P = 0.007) were independent prognostic factors. A prognostic index was developed which divided the patients into different risk groups. It was shown that high-risk patients identified by this index were mainly found in the group prescribed nonprotocol treatment and that the prognostic index could not be used to identify patients with a low probability of response among patients prescribed protocol treatment. PMID- 8423021 TI - Molecular genetic evidence of a unifocal origin for human serous ovarian carcinomas. AB - The hypothesis that ovarian cancer is multifocal in origin was examined using molecular genetic techniques. Patterns of allelic deletion on chromosome 17 were studied in 16 informative cases of Stage III serous epithelial ovarian carcinoma. DNA was extracted from specimens collected from the omentum and both ovaries, and the specific alleles and chromosomal loci involved in the deletion were identified and compared. In all cases, the patterns of allelic deletion were identical for the tumors that had been collected from different sites in the same patients. In addition, 4 of the 16 cases were heterozygous for the hypoxanthine phosphoribosyl transferase (HPRT) gene on the X-chromosome and were examined for methylation status. In all 4, the same parental allele of the HPRT gene was methylated in tumor cells collected from both ovarian and omental sites, suggesting that the patterns of inactivation of the X-chromosome are identical. This pattern of allelic deletion and HPRT-gene methylation in tumor samples collected from different sites implies that ovarian carcinomas have a unifocal origin. PMID- 8423022 TI - Determinants of invasive vulvar cancer risk: an Italian case-control study. AB - Risk factors for vulvar cancer have been evaluated in a case-control study conducted between 1987 and 1990 in northern Italy on 73 women with histologically confirmed invasive vulvar cancer and 572 control subjects in hospital for acute nongynecological, nonneoplastic non-hormone-related conditions. The risk of vulvar cancer was inversely related to education level: with reference to women reporting less than 7 years of schooling, the relative risk estimates were 0.6 and 0.4, respectively, in those reporting 7 to 11 and 12 or more years of schooling (chi 2(1) trend = 4.91 P = 0.03). No relationship emerged between number of births and spontaneous or induced abortions. Parous women reporting late first birth tended to be at lower risk (relative risk = 0.5, 95% confidence interval 0.3 to 1.1 for < 25 vs > or = 25 years at first birth), but there was no evidence of the risk to decrease with increasing age at first birth. The risk of vulvar cancer increased with body mass index, but the trend in risk was not significant after taking into account potential confounders in the multivariate analysis. No association emerged with indicators of sexual habits, menstrual history, and smoking. The risk of the disease was lower in women reporting Pap smears during their life and diminished with increasing number of cervical smears and decreasing recency of last Pap: compared to women reporting no Pap screening, it was 0.5 in those who reported one smear and 0.3 in those with two or more. PMID- 8423023 TI - High-resolution positron emission tomography of human ovarian cancer in nude rats using 124I-labeled monoclonal antibodies. AB - PET has inherently high resolution and excellent contrast imaging and accurately measures radioactivity concentrations in vivo. When combined with specific immunological targeting it might provide a highly specific and sensitive radioimmunoscintigraphic tool. To investigate this we injected 124I-labeled MAb MX35 or MAb MH99 monoclonal antibodies (doses 200-400 mu Ci) intravenously into nude rats bearing subcutaneous human ovarian cancer xenografts (SK-OV-7 and SK-OV 3 cell lines). A melanoma cell line (SK-MEL-30) was used as a control tumor. These murine monoclonal antibodies react with cell-surface antigens expressed by most ovarian cancer cells, including the ovarian cell line used. Imaging was performed at 1-6 days using a high-resolution positron emission tomograph (PCR-I) with a spatial resolution of 4.5 mm. The slice thicknesses were 0.5 and 1.0 cm. Forty to seventy thousand coincident pulses were obtained per frame. The PET results were compared with those of autopsy and histology. Samples of blood, tumor, and normal tissues were obtained at various time points. PET calculation of isotope uptake ratios demonstrated specific localization of the antibodies in tumor, with ratios of tumor to normal tissue uptake as high as 6:1. Subcutaneous ovarian cancer nodules as small as 7 mm were identified with PET imaging. The results corresponded well with tissue sampling. Our findings suggest that PET imaging of tumors with 124I-labeled monoclonal antibodies may be useful in human diagnostic and therapeutic applications in ovarian cancer as well as other diseases. PMID- 8423024 TI - Surgical management of early invasive cancer of the cervix associated with pregnancy. AB - The simultaneous occurrence of carcinoma of the cervix in pregnancy is uncommon. In a prospective study of 397 patients undergoing type III radical hysterectomy for early invasive cancer of the cervix, 18 were pregnant at the time of surgery; 4 of these were operated after delivery elsewhere. Two others had a type I extrafascial hysterectomy. The incidence was 1 in 4077 deliveries. The clinical and histological characteristics of these patients are presented. Routine speculum examination and cervical cytology in all pregnant patients early in pregnancy are vital for early diagnosis. Bleeding in pregnancy should not automatically be assumed to be caused by pregnancy-related conditions. The strategies in surgical management are discussed. Radical hysterectomy in pregnancy is safe. No major complications were encountered; the mean blood loss was 1.4 liters. The incidence of pelvic node metastases was similar to that seen in nonpregnant patients. The overall 5-year survival rate was 77.7%. Of the 4 patients who died, 3 presented in the puerperium; all succumbed within 27 months. Poor histological prognostic features contributed to the significantly poorer survival in the puerperal patients (P = 0.0445). The 5-year survival in those presenting during the antepartum period (92.8%) was, however, similar to that in the nonpregnant patients. Metastases to the placenta or fetus were not encountered. PMID- 8423026 TI - Squamous cell carcinoma of the vagina: a review of 70 cases. AB - Seventy cases of squamous cell carcinoma of the vagina registered between 1985 and 1989 were analyzed. The overall 2-year disease-free survival was 33%. Stagewise 2-year survivals were as follows: stage I, 100% (8/8); stage II, 70% (7/10); stage III, 19% (8/42); and stage IV, 0% (0/10). Incidentally, 60% of the cases presented below 50 years of age, and the majority of these were in advanced stage (p < 0.02). Due to the relatively greater number of cases of advanced disease in the elderly age group treated with external radiotherapy alone, a significant survival difference between the two age groups was not apparent (P > 0.10). External radiotherapy alone yielded poor results. External radiotherapy in combination with brachytherapy in the form of either vaginal cylinders or uterine tandems with vaginal cylinders resulted in 42 and 50% 2-year disease-free survivals, respectively. Advanced stage, more than two-thirds to full vaginal length involvement, and multiple vaginal wall involvement were found to be poor prognostic factors. The majority of cases had tumor grades 2 and 3. No association between tumor grade and survival was observed. To improve survival, downstaging of the disease through routine gynecological checkup, even in premenopausal women, and delivery of high doses through a judicious combination of external radiotherapy and brachytherapy are needed. PMID- 8423025 TI - Ketorolac tromethamine, a nonsteroidal anti-inflammatory drug: ability to inhibit post-radical pelvic surgery adhesions in a porcine model. AB - The objective of this study was to investigate the ability of ketorolac tromethamine, a parenterally administered nonsteroidal anti-inflammatory drug (NSAID), to inhibit post-radical pelvic surgery adhesion formation (PRPSA). Twenty adult female hogs undergoing radical hysterectomy, bilateral salpingo oophorectomy, and resection of pelvic peritoneum were used. Animals were randomized to either ketorolac (4 mg/kg loading dose 45 min prior to skin incision, followed by 2 mg/kg IM every 8 hr for nine doses) or no adjuvant therapy. Four weeks after surgery, repeat celiotomy was performed, and adhesions were quantified. Note was made of any bowel segments adherent to the pelvis. Adhesion scores for the ketorolac-treated animals (n = 9, mean = 0.92 +/- 0.54, median = 0.90) were significantly less than those for control animals (n = 10, mean = 6.73 +/- 2.21, median = 7.03, P = 0.0001). Similarly, significantly fewer treated animals had small bowel loops adherent in the pelvis when compared to control animals (11% vs 90%, P = 0.001). Ketorolac is an effective inhibitor of PRPSA formation and limits small bowel adherence to the pelvis. PMID- 8423027 TI - Estro-progestinic replacement therapy modulates the levels of basic fibroblast growth factor (bFGF) in postmenopausal endometrium. AB - The levels of basic fibroblast growth factor (bFGF) in human endometrium do not vary during the ovulatory cycle but they increase significantly after menopause (M. Rusnati et al., Growth Factors, 3, 299-307, 1990). Here we report that cyclic estro-progestinic replacement therapy reduces the levels of bFGF in human postmenopausal endometrium to those measured in ovulatory cycle endometrium. Also, we have observed that the levels of bFGF in well-differentiated adenocarcinomas of the endometrium from postmenopausal patients are lower than those detected in their normal counterpart and are comparable to those measured in the normal cycling tissue. The data indicate that the stimulation of cell proliferation induced in postmenopausal endometrium by hormonal therapy or by neoplastic transformation is paralleled by a decrease of endometrial levels of bFGF. This may be due to an increase of the turnover rate of the growth factor in the proliferating versus the quiescent tissue and suggests a role of bFGF in the growth of normal and neoplastic human endometrium. Our data represent the first evidence that sex hormones modulate the levels of biologically active bFGF in vivo. PMID- 8423028 TI - Nuclear type II estradiol binding sites and type I estrogen receptors in human endometrial cancer: a 5-year follow-up study. AB - Type II nuclear-estrogen binding sites (type II EBS) have been postulated to play a crucial regulatory role in estrogen-stimulated uterine growth. The pathogenesis of endometrial cancer is known to be connected to estrogens. However, there are no data on type II EBS in human endometrial cancer. Therefore, we investigated the presence of nuclear type II EBS and nuclear type I estrogen receptors (ER) by radioligand-binding assays in endometrial tumorous tissues (n = 135). Determination of cytoplasmic ER and progesterone receptor (PR) concentrations were also included in the study. Estradiol (E2) and progesterone (P) hormone levels in sera of patients were determined by RIAs. In addition, all data were analyzed on 5-year survival rates. Saturation analyses of 3H-E2 binding in crude nuclear pellets (n = 5) showed two types of estradiol binding: type I ER having high affinity and low capacity and type II EBS binding estradiol with lower affinity and high capacity in a positive cooperative fashion. The nuclear type II EBS and type I ER concentrations were significantly higher in cancers of increasing grade. In contrast, a significant decrease of cytoplasmic 3H-E2 binding was detected. Cytoplasmic PR binding capacities were high in G1 and G2, but low in G3 tumors. The 5-year survival data showed nuclear type I ER to have the best correlation for prognostic value (cut-off, 0.5 pmol/mg DNA), while type II EBS had no significant impact on it. Serum E2 concentrations decreased with tumors of higher grade, but were generally still higher than the control values. The serum P levels did not alter. None of the parameters investigated here differed from control values in adenoacanthoma (n = 6), suggesting a separate pathomechanism. As type II EBS are known to be stimulated by E2 under conditions that cause uterine hyperplasia, we concluded that the higher "runaway" nuclear type II EBS levels, in contrast with the lower serum estradiol concentrations, might be connected with the strong proliferative and invasive character of higher grade endometrial adenocarcinomas. This is the first demonstration of the presence of nuclear type II EBS and their possible pathological role in human endometrial cancer, and of the high prognostic significance of nuclear type I ER to identify a subgroup having a fatal form of the disease in a 5-year survival study. PMID- 8423029 TI - [Onychomycoses--therapeutic challenge for physician and patient. Report from the MYK 92 26th Scientific Meeting of the German-speaking Mycology Association. Graz, 4 September 1992]. PMID- 8423030 TI - Pregnancy and cholelithiasis: pathogenesis and natural course of gallstones diagnosed in early puerperium. AB - Several recent reports have indicated an increased prevalence of gallstones in association with pregnancy. If these reports are true, the early puerperium should be a favorable time to detect the disease in its initial stages and follow its natural course. Accordingly, the gallbladder was examined by ultrasound in 980 women during the immediate postpartum period and in 150 nulliparous, age matched healthy volunteers. Gallstones were detected in 12.2% of the puerperal women and in 1.3% of the control group. In 70 patients who had stones in a functioning gallbladder, 22 (31%) had had attacks of biliary colic. The history of pain was more common in patients with stones greater than 10 mm in diameter. Forty-one women with small stones (< 10 mm) were followed clinically and ultrasonographically for between 6 and 24 (mean = 8.7) mo. All remained pain free, and in twelve subjects (29%) the stones disappeared. Gallbladder bile was examined in 11 normal volunteers (controls) immediately after delivery and in 19 women with small stones 39 +/- 6 days postpartum. Bile was saturated with cholesterol in the controls and was unsaturated in patients with gallstones. We conclude that in our population pregnancy is a very important pathogenetic factor favoring gallstone formation. Attacks of biliary colic appear early and frequently in young Chilean women with this disease. Unexplained disappearance of small stones frequently occurs: in some cases it is likely to be the result of spontaneous dissolution because bile becomes unsaturated within a few weeks of delivery. PMID- 8423031 TI - Bile acid sulfonates alter cholesterol gallstone incidence in hamsters. AB - The prevention of cholesterol gallstone formation by three bile acid analogs, sodium 3 alpha,7 alpha-dihydroxy-5 beta-cholane-24- sulfonate, sodium 3 alpha,7 beta-dihydroxy-5 beta-cholane-24-sulfonate and sodium 3 alpha,6 alpha-dihydroxy-5 beta-cholane-24-sulfonate, was examined in a hamster model of cholesterol cholelithiasis. Sodium taurochenodeoxycholate, sodium tauroursodeoxycholate and sodium taurohyodeoxycholate were studied simultaneously for comparison. Gallstones and cholesterol crystals were induced in 14 of 15 hamsters fed a bile acid-free, semipurified lithogenic diet containing 0.3% cholesterol and 4% butterfat for 6 wk. The addition of 0.1% sodium taurochenodeoxycholate and sodium tauroursodeoxycholate to the lithogenic diet had little effect on the formation of gallstones or biliary cholesterol crystals. In contrast, sodium 3 alpha,7 alpha-hydroxy-5 beta-cholane-24- sulfonate and sodium 3 alpha,7 beta-dihydroxy-5 beta-cholane-24-sulfonate, when fed at the same dose, prevented cholesterol gallstone formation significantly. Sodium taurohyodeoxycholate and sodium 3 alpha,6 alpha-dihydroxy-5 beta-cholane- 24-sulfonate inhibited cholesterol gallstone formation effectively. The cholesterol saturation index of bile was greater than 1.00 in all groups, with the exception of the group fed sodium 3 alpha,7 alpha-dihydroxy-5 beta-cholane-24-sulfonate. Liver and serum cholesterol levels tended to be lower in most of the groups that were fed bile acids. This effect was most pronounced in the animals receiving sodium taurohyodeoxycholate. At the end of the experiment, the administered sulfonate analogs were detected in gallbladder bile.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423032 TI - Biliary copper excretion in acutely and chronically copper-loaded rats. AB - Biliary copper excretion was examined in rats with acute, continuous and chronic copper loads. Copper was excreted into bile, and the concentration peaked 40 min after a venous injection of copper sulfate (127 ng/gm body weight). The excretion was significantly inhibited by colchicine. Therefore some copper may be transported in hepatocytes by a vesicular pathway and excreted into bile. Biliary copper output increased over time and reached a plateau 180 min after a continuous venous infusion of copper sulfate (318 ng/gm body weight/hr) had started, when the concentration of copper in bile was much higher than that in plasma; the bile/plasma ratio of copper concentrations was 4.32 +/- 0.46. These data support the idea that copper transport involves a specific uptake and transport system. In chronically copper-loaded rats, hepatic copper content was significantly increased compared with controls, and reaction products for copper were observed in hepatocyte granules by light microscopic examination with p dimethylaminobenzylidene rhodanine stain. The number of lysosomes in hepatocytes increased and the shape changed. In chronically copper-loaded rats the number of tubular lysosomes was very high. However, other organelles appeared to be normal. In these rats biliary excretion of not only copper but also acid phosphatase, a lysosomal enzyme, was significantly greater than the control. Therefore hepatocyte lysosomes may play an important role in biliary copper excretion. Furthermore, when biliary lysosomal excretion increases, the tubular lysosomes actively participate in this excretion. PMID- 8423033 TI - The clearance rate of chylomicron retinyl ester from plasma can be used to distinguish rats with cirrhosis from those with portacaval shunt. AB - Effects of carbon tetrachloride treatment and portacaval shunt surgery on exogenous chylomicron retinyl ester clearance from rat plasma were analyzed assuming three-compartment model kinetics. In rats with cirrhosis and in those with Eck fistulas, the relative pool size of compartment 2 decreased (0.20 and 0.36, respectively) compared with controls (0.82). The relative mass of compartment 3 significantly increased in rats with cirrhosis (1.55) or Eck fistula (0.19) compared with control rats (0.11). The cirrhotic and Eck fistula groups were indistinguishable on the basis of these parameters and on the basis of indocyanine green test values. However, the cirrhosis and Eck fistula groups differed clearly from each other with respect to fractional efflux rate constants (l2, l3), where the constant l2 is from compartment 2 and the l3 is from compartment 3. Both values decreased in cirrhotic rats, suggesting that the hepatic uptake of chylomicron retinyl esters was impaired by carbon tetrachloride. On the other hand, Eck fistula rats did experience dramatic increases in l3, implying that the hepatic uptake of chylomicron retinyl esters from compartment 3 was enhanced by portacaval shunting. Elevation of the plasma estrogen level observed in Eck fistula rats may be responsible for the induction of low-density lipoprotein receptors on hepatocytes and for the subsequent enhancement of l3. These results suggest that a three-compartment model of plasma retinyl ester disappearance kinetics gives important quantitative information about hepatic function. Clinical application of the chylomicron retinyl ester clearance test is discussed for estimating hepatic function reserves and for differential diagnosis of portal hypertension. PMID- 8423034 TI - Experimental hepatitis induced by ethanol after immunization with acetaldehyde adducts. AB - We produced hepatitis in guinea pigs by immunization with acetaldehyde adducts and ethanol treatment. Human hemoglobin-acetaldehyde adducts were prepared without any reducing agents and affinity purified with polyclonal antibodies against acetaldehyde adducts. Female guinea pigs were immunized with the adducts and were simultaneously given ethanol for 40 days. These treatments induced hepatic necrosis with infiltration of mononuclear cells in the hepatic lobules. The formation of the lymphoid follicle was also observed in severe cases. These changes were accompanied by the elevation of serum AST and lactic dehydrogenase activities and titers of circulating antibodies against acetaldehyde adducts. By contrast, the combination of ethanol and immunization with unmodified hemoglobin produced only fatty change of the liver, and animals immunized with the adducts alone had minimal inflammatory changes of the liver. Peripheral blood lymphocytes obtained from the animals with hepatitis were shown to be stimulated by acetaldehyde adducts to a significantly greater degree than those from control animals who received nothing, ethanol alone or ethanol and unmodified hemoglobin. These results suggest that the immune response to acetaldehyde adducts may be involved, at least partly, in the pathogenesis of inflammation observed in alcoholic liver disease. PMID- 8423035 TI - Vasopressin gene expression in rats with experimental cirrhosis. AB - Impaired ability to excrete a water load occurs in a substantial number of patients with advanced cirrhosis and in animals with experimental cirrhosis. The nonosmotic stimulation of arginine vasopressin release from the pituitary has been implicated as an important factor in the abnormal water excretion in patients and animals with cirrhosis. In this study, arginine vasopressin hypothalamic gene expression was studied in cirrhotic rats. Cirrhosis was induced by a combination of phenobarbital treatment in drinking water and weekly intragastric administration of carbon tetrachloride for 13 to 15 wk. Severe cirrhosis was confirmed by morphological analysis and the presence of ascites. Plasma arginine vasopressin was also significantly higher in rats with cirrhosis (control = 1.77 +/- 0.16 and cirrhotic rats = 4.14 +/- 0.62 pg/ml, n = 9, p < 0.002). Hypothalamic arginine vasopressin messenger RNA was also significantly higher in cirrhotic rats (control = 762.1 +/- 132.3 and cirrhotic rats = 1,834.2 +/- 271.9 pg/hypothalamus, n = 9, p < 0.005). Pituitary arginine vasopressin content was significantly lowered in cirrhotic rats (control = 3.69 +/- 0.98 and cirrhotic rats = 1.57 +/- 0.09 micrograms/pituitary, n = 9, p < 0.05). No difference was seen in hypothalamic arginine vasopressin content between the two groups (control = 4.64 +/- 0.34 and cirrhotic rats = 4.23 +/- 0.33 ng/hypothalamus, n = 9, NS). Oxytocin messenger RNA in the hypothalamus was also not significantly different between the two groups (control = 8.61 +/- 0.68 and cirrhotic rats = 9.33 +/- 0.65 unit of density, n = 9, NS).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423036 TI - Transjugular intrahepatic portal-systemic shunts: the state of the art. PMID- 8423037 TI - Pregnancy and gallstones. PMID- 8423038 TI - Transforming growth factor-beta 1: there is regulation beyond transcription. PMID- 8423039 TI - Differentiation between heterozygotes and homozygotes in genetic hemochromatosis by means of a histological hepatic iron index: a study of 192 cases. AB - The biochemical hepatic iron index, defined as the ratio of hepatic iron concentration (expressed as micromoles per gram dry weight) to age permits accurate prediction of genetic status in patients with genetic hemochromatosis. However, the hepatic iron concentration is not always available. Therefore a histological hepatic iron index, defined as the ratio of total histological iron score (range = 0 to 60) to age, was evaluated in a total of 192 Australian and French patients with genetic hemochromatosis. These subjects had been classified previously as heterozygotes (n = 18) or homozygotes (n = 174) according to clinical and familial data only. Biochemical hepatic iron index and histological hepatic iron index were well correlated (Spearman's test: rho = 0.75, p < 0.0001). Both were significantly (p < 0.0001) increased in homozygotes (respectively, 6.7 +/- 3.8 [range = 1.2 to 22.6] and 0.62 +/- 0.28 [range = 0.14 to 1.5]) compared with heterozygotes (respectively, 1 +/- 0.4 [range = 0.45 to 1.6] and 0.08 +/- 0.05 [range = 0 to 0.14]). The histological hepatic iron index was less than 0.15 in all heterozygotes and greater than 0.15 in all but two homozygotes. These data show that the age-dependent nature of iron accumulation can also be accommodated by calculating the histological hepatic iron index and that histological study is an accurate means of predicting the genetic status of hemochromatosis patients when hepatic iron concentration is not available. PMID- 8423040 TI - Orthotopic liver transplantation in two adults with Niemann-Pick and Gaucher's diseases: implications for the treatment of inherited metabolic disease. AB - Two adults were seen with cirrhosis caused by different lipid storage diseases. A 42-yr-old woman with Niemann-Pick disease type B had marked hepatomegaly, ascites and recent variceal bleeding. Her evaluation showed chronic bilateral pulmonary infiltrates, multiple stigmata of chronic liver disease including the recent cessation of menses, diuretic-resistant sterile ascites, hepatic encephalopathy and variceal bleeding. Five percent of normal sphingomyelinase activity was measured in peripheral leukocytes. A 42-yr-old man with Gaucher's disease and a history of bilateral hip replacements presented with hepatomegaly, jaundice, refractory ascites and renal insufficiency. His evaluation showed 20% to 23% of normal glucocerebrosidase activity in peripheral leukocytes. Both patients underwent orthotopic liver transplantation with resolution of all aspects of decompensated liver function. Assessment of the underlying metabolic defect before and 6 to 14 mo after transplantation showed that after transplantation the patient with Niemann-Pick disease had above normal hepatic sphingomyelinase activity, a less-marked increase in peripheral leukocyte enzyme activity and lower than normal hepatic sphingomyelin and cholesterol content. In contrast, the patient with Gaucher's disease had only a 61% increase in hepatic glucocerebrosidase activity but had an elevated hepatic glucocerebroside content that was only 15% of the pretransplant level and decreased peripheral leukocyte enzyme levels. These findings suggest that variable relationships may exist between posttransplant hepatic and peripheral leukocyte enzyme activities in the different lipidoses, which may have implications for recurrence of glycolipid induced liver damage. PMID- 8423041 TI - Oddi's sphincter motor activity in patients with recurrent pyogenic cholangitis. AB - Oddi's sphincter motor activity was studied in 15 healthy subjects and 15 patients with recurrent pyogenic cholangitis. No significant difference was found in the common bile duct pressures, the Oddi's sphincter basal pressures or the amplitude, frequency and duration of phasic contractions between the controls and patients. However, a significant difference was found in the percentage of antegrade and retrograde phasic wave sequences between the two groups. In control patients 64.0% +/- 5.3% of phasic wave sequences were antegrade, and 20% +/- 3.6% were retrograde. In patients with recurrent pyogenic cholangitis, 37% +/- 3.6% of phasic wave sequences were antegrade, and 42% +/- 5.5% were retrograde. No significant difference was found in the percentage of simultaneous phasic wave sequences (16% +/- 1.5% vs. 18.5% +/- 2.8%, respectively) between the two groups. In all, five (33%) patients with recurrent pyogenic cholangitis had abnormal propagation of phasic contractions of Oddi's sphincter (retrograde phasic wave sequences greater than 50%), two (13%) patients had elevated Oddi's sphincter basal pressures (greater than 40 mm Hg) and one (7%) patient had increased frequency of Oddi's sphincter phasic contractions (more than 9/min). Six of the 15 patients had evidence of papillitis on duodenoscopy. We believe papillitis could induce motor abnormalities in these patients, thus resulting in delayed biliary drainage and recurrent attacks of cholangitis. PMID- 8423042 TI - Effect of dipyridamole on kidney function in cirrhosis. AB - Adenosine is a potent endogenous renal vasoconstrictor. To investigate the sensitivity of the renal circulation to adenosine in cirrhosis, we evaluated kidney function and vasoactive hormones in 20 patients with cirrhosis before and after administration of dipyridamole (0.4 mg/kg, intravenously), a drug that increases extracellular levels of adenosine. In patients with ascites and increased plasma renin activity (6.9 +/- 4.0 ng/ml.hr [mean +/- S.D.]) (n = 7), dipyridamole induced marked reductions in renal plasma flow (from 623 +/- 294 to 374 +/- 188 ml/min, p = 0.03), glomerular filtration rate (from 89 +/- 22 to 48 +/- 16 ml/min, p = 0.009), urine volume (from 7.1 +/- 2.1 to 1.5 +/- 1.1 ml/min, p = 0.0001), free water clearance (from 4.0 +/- 1.7 to 0.4 +/- 0.6 ml/min, p = 0.001) and sodium excretion (from 28 +/- 36 to 7 +/- 15 mu Eq/min, p = 0.05) in the absence of changes in arterial pressure, plasma renin activity and levels of aldosterone, norepinephrine and antidiuretic hormone. In patients without ascites (n = 5) and in patients with ascites and normal plasma renin activity (0.9 +/- 0.5 ng/ml.hr) (n = 8), renal plasma flow and glomerular filtration rate did not change significantly after dipyridamole administration, whereas excretion of sodium and free water was reduced. These results indicate that in cirrhotic patients with ascites and overactivity of the renin-angiotensin system, dipyridamole induces renal vasoconstriction in the absence of changes in systemic hemodynamics, suggesting that these patients are particularly sensitive to the renal vasoconstrictor effect of endogenous adenosine. PMID- 8423043 TI - Depression of drug-metabolizing activity in the human liver by interferon-beta. AB - The depressant effect of interferon beta on drug-metabolizing activity in the human liver was investigated. Seven patients with chronic hepatitis C were treated with interferon beta at doses of 3 x 10(6) to 9 x 10(6) IU/day for 8 wk. The activities of 7-methoxycoumarin O-demethylase and 7-ethoxycoumarin O deethylase in specimens obtained by liver biopsy were examined before and after interferon treatment. Theophylline pharmacokinetics were also examined before and after interferon treatment. Interferon beta treatment reduced the activities of both O-dealkylases from 6.0 (100%) to 3.2 (53%) nmol/gm liver per minute and from 1.9 (100%) to 1.1 (58%) nmol/gm liver per minute, respectively (p < 0.05). The total body clearance of theophylline was also decreased (from 0.76 to 0.56 ml/kg/min; p < 0.05), and its elimination half-life was increased (from 8.4 to 11.7 hr; p < 0.05); however, the volume of distribution was not significantly affected. The magnitude of the decreases in enzyme activities and in theophylline clearance varied widely in individual patients and did not correlate with the dose of interferon administered. This study provides the first direct evidence that interferon beta can depress the activity of drug-metabolizing enzymes in the human liver. PMID- 8423044 TI - Hyperfibrinolysis resulting from clotting activation in patients with different degrees of cirrhosis. The CALC Group. Coagulation Abnormalities in Liver Cirrhosis. AB - This study explored the relationship between clotting activation and tissue plasminogen activator and its inhibitor in cirrhotic patients with different degrees of liver failure. Sixty-seven patients (40 men, 27 women; age = 31-77 yr) with cirrhosis diagnosed by liver biopsy were divided into three subgroups (A, B and C) on the basis of Child-Pugh classification. Tissue plasminogen activator antigen and activity, plasminogen activator inhibitor antigen and activity, fibrin/fibrinogen degradation products, and D-dimer were measured in each patient. Forty-two patients with normal levels of fibrin/fibrinogen degradation products and D-dimer showed significant progressive decreases of plasminogen activator inhibitor antigen levels (p < 0.01) and activity (p < 0.0001) from class A to class C. This decrease was significantly related to prothrombin time (p < 0.003). Tissue plasminogen activator values were not different in the three Child classes. Twenty-five patients (7 class B and 18 class C) with high circulating values of fibrin/fibrinogen degradation products and D-dimer had higher values of tissue plasminogen activator antigen (20.0 +/- 10.1 ng/ml vs. 5.9 +/- 3.0 ng/ml; p < 0.0001) and activity (6.9 +/- 2.2 U/ml vs. 2.1 +/- 1.3 U/ml; p < 0.0001) and lower values of plasminogen activator inhibitor antigen (6.9 +/- 4.1 ng/ml vs. 14.8 +/- 5.6 ng/ml; p < 0.0001) and activity (4.1 +/- 2.8 U/ml vs. 9.8 +/- 3.7 U/ml; p < 0.0001) than did patients with normal values of fibrin/fibrinogen degradation products and D-dimer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423045 TI - N omega-nitro-L-arginine administration corrects peripheral vasodilation and systemic capillary hypotension and ameliorates plasma volume expansion and sodium retention in portal hypertensive rats. AB - In portal hypertensive states, peripheral vasodilation leads to sodium retention and plasma volume expansion. N omega-nitro-L-arginine, a specific biosynthesis inhibitor of the vasodilator nitric oxide, has been shown to acutely reverse peripheral vasodilation and the vascular hyporesponsiveness to endogenous and exogenous vasoconstrictors observed in portal hypertensive rats. This study investigated whether N omega-nitro-L-arginine treatment in portal hypertensive rats prevents peripheral vasodilation and therefore ameliorates plasma volume expansion and sodium retention. For 2 days before partial portal vein ligation or sham operation and then continuously for 4 days after the operation, animals received either placebo (0.9% saline) or N omega-nitro-L-arginine (approximately 2 micrograms/kg/min) intravenously through a subcutaneously implanted Alzet osmotic pump (model 2ML1; Alza, Palo Alto, CA). In portal hypertensive rats, N omega-nitro-L-arginine treatment significantly increased mean arterial pressure (placebo vs. N omega-nitro-L-arginine, 123 +/- 4 vs. 150 +/- 2 mm Hg, respectively; p < 0.001) and systemic vascular resistance (3.8 +/- 0.2 vs. 5.6 +/ 0.3 mm Hg/ml/min/100 gm body weight; p < 0.001), associated with a decrease in the cardiac index (33.5 +/- 1.0 vs. 27.0 +/- 1.1 ml/min/100 gm body weight; p < 0.001). N omega-nitro-L-arginine treatment also induced a decrease in plasma volume (4.6 +/- 0.1 vs. 4.1 +/- 0.1 ml/100 gm body weight; p < 0.001) and extracellular sodium space (39.4 +/- 0.7 vs. 37.4 +/- 0.4 ml/100 gm body weight; p < 0.05) without changes in serum sodium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423046 TI - Glycine protects hepatocytes from injury caused by anoxia, cold ischemia and mitochondrial inhibitors, but not injury caused by calcium ionophores or oxidative stress. AB - Isolated hepatocytes, suspended in an organ preservation solution, can be preserved at 4 degrees C for up to 6 days. After preservation, normothermic normoxic incubation causes loss of hepatocyte viability. The addition of 3 mmol/L glycine to the rewarming medium prevents the loss of viability. In this study we investigated the cytoprotective effects of glycine under many conditions known to cause hepatocellular injury to understand the mechanism of cold-induced injury in the liver. Hepatocytes were suspended in modified Krebs-Henseleit buffer with or without 3 mmol/L glycine and exposed to agents or conditions known to induce cell death. Hepatocyte viability was assessed by measuring the percentage of lactate dehydrogenase leakage from the cells and the concentration of ATP during incubation at 37 degrees C under room air for up to 90 min. Mitochondrial inhibitors (potassium cyanide and carbonyl cyanide m-chlorophenylhydrazone); calcium ionophores (ionomycin and A23187); an oxidizing agent, tert-butyl hydroperoxide; and anoxia were all used to cause cell injury. Hepatocytes were also isolated from fasted rats and hypothermically preserved as another model of cell death. Other amino acids were also tested in the hypothermic preservation model to study the specificity of the amino acid requirement for prevention of lactate dehydrogenase leakage. Of the amino acids tested, only alanine (10 mmol/L) and the combination of alanine (3 mmol/L) and serine (3 mmol/L) were as effective as glycine in preventing lactate dehydrogenase release in the hypothermic preservation model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423047 TI - Increase in ornithine decarboxylase activity caused by hepatocyte growth factor in primary cultured adult rat hepatocytes. AB - The effect of hepatocyte growth factor on ornithine decarboxylase activity was studied in primary cultured adult rat hepatocytes. Ornithine decarboxylase activity was increased 3 hr after the addition of hepatocyte growth factor and remained at a high level until 12 hr; thereafter it decreased, and it returned to the control level by 24 hr. Enzyme activity began to increase with 1 ng/ml hepatocyte growth factor and reached its maximum with 5 ng/ml hepatocyte growth factor. When insulin or epidermal growth factor was added with hepatocyte growth factor, enzyme activity was further stimulated. The level of ornithine decarboxylase messenger RNA did not increase with addition of hepatocyte growth factor. The half-time of ornithine decarboxylase activity was prolonged about twofold by hepatocyte growth factor treatment. These results suggest that hepatocyte growth factor treatment of cells enhanced ornithine decarboxylase activity posttranslationally. PMID- 8423048 TI - Genomic structure of NKG5, a human NK and T cell-specific activation gene. AB - We reported previously the isolation of a cDNA clone, designated NKG5, encoding a secreted protein that is expressed only in natural killer and T cells and is strongly upregulated upon cell activation. In this report we have isolated the NKG5 gene from a human placental genomic library and sequenced the gene and two kilobases of 5'-flanking DNA. Comparison with the cDNA sequence reveals that the NKG5 gene consists of five exons and four introns. Intron 1 contains a DNA segment that was reported to occur as an exon in 519, a closely related cDNA clone that was isolated from a T-cell library. This result indicates that NKG5 and 519 are alternative splicing products of a single gene. The 5'-flanking region of the NKG5 gene was analyzed for homology with the promoter regions of cytokines and other activation-induced genes showing lymphocyte-specific expression. Several segments displaying sequence similarity were identified. We also identified numerous sequence elements that have strong similarity to known binding sites for transcriptional regulatory proteins including T cell-specific and activation-specific regulatory factors. These findings are consistent with the cell-specific expression and the tight regulatory control that is observed for the NKG5 gene. PMID- 8423049 TI - Allelic variations clustered in the antigen binding sites of HLA-Bw62 molecules. AB - HLA-Bw62 is a serologically defined class I antigen specificity, but we show that it represents a family of five distinct alleles in this study. Five variants of HLA-Bw62 antigens were identified by isoelectric focusing, and sequencing studies revealed that these are a family of closely related alleles differing from one another by one to six amino acid substitutions at eight positions: 63 in the alpha 1 domain and 94, 95, 97, 99, 113, 152, and 156 in the alpha 2 domain. These substitutions are located in the two alpha-helices and two adjacent beta-strands, and the side chains of most amino acids face into the antigen binding groove. Functional assays using an in vitro generated Epstein-Barr virus (EBV)-specific Bw62-restricted cytotoxic T lymphocyte clone indicated that the minimal structural variations located in the antigen binding sites of the HLA-Bw62 variant molecules could affect the presentation of the nominal EBV antigen. This study revealed that the HLA-Bw62 antigen family consists of at least five closely related alleles, and further demonstrated that these alleles with minimal structural variations might play distinct functional roles in regard to antigen presentation. PMID- 8423050 TI - The thioester and isotypic sites of complement component C4 in sheep and cattle. AB - The region inclusive of the thioester and the isotype-determining sites of the sheep C4 genes from a single animal was amplified by polymerase chain reaction (PCR). Two bands, at 880 base pairs (bp) and 1000 bp, were resolved by agarose gel electrophoresis. Four different clones were obtained for the 880 bp (type 1) product and two from the 1000 bp (type 2) product. Two of the type 1 clones (type 1H) and both type 2 clones (type 2H) code for the PCPVIH sequence at the isotypic site whereas the other two type 1 clones (type 1D) code for the PFPVMD sequence. By restriction mapping and Southern blot analysis, there appears to be four C4 gene loci for the sheep: two type 1H, one type 1D, and one type 2H. The type 1H and type 2H genes are likely to code for proteins with C4B-like properties whereas the type 1D genes for proteins with C4A-like properties. The same region of the sheep C4 genes of nine other breeds of sheep are also amplified by PCR and analyzed by restriction mapping and Southern hybridization. Each of the sheep has type 1H, type 2H, and type 1D genes and appears to have four C4 gene loci except for the Orkney, which may have five. A single band of 880 bp was obtained from the PCR product from the genomic DNA of a single cow. Five different clones were identified, two of which code for the PFPVMD sequence and three for the PCPVIH sequence at the isotypic site, which is consistent with previous finding that C4 proteins with A- and B-like activities could be purified from the plasma of the same animal. Comparison of the nucleotide sequences of the isotype-determining region of the sheep and cattle C4 genes with those of the primates and mouse suggests that the C4A-like genes evolved independently in the primates and the ungulates. PMID- 8423052 TI - Tum- mutation P35B generates the MHC-binding site of a new antigenic peptide. AB - Immunogenic tumor cell variant P35 was obtained by mutagen treatment of mouse mastocytoma P815. It expresses a potent new antigen recognized by syngeneic cytolytic T lymphocytes (CTL). This antigen is the result of a point mutation in a gene that is expressed by most healthy cells. A decapeptide encoded by the region spanning the mutation sensitized P815 cells to the relevant CTL, whereas the homologous decapeptide corresponding to the normal sequence did not. Only the mutant decapeptide was capable of enhancing the expression of the Dd-presenting molecule at the cell surface, indicating that the mutation generates a motif which enables the antigenic peptide to bind to Dd. PMID- 8423051 TI - Differential susceptibility of mouse Lyt-2 and Lyt-3 genes to negative regulation. AB - Hybrids of Lyt-2/Lyt-3-positive class I-specific cytotoxic T lymphocytes (CTLs) with the BW5147 thymoma cell line (Lyt-2/Lyt-3-negative) are known to be Lyt 2/Lyt-3-negative due to shutoff of transcription of the CTL's Lyt-2 gene. Hybrids of a constitutively Lyt-2-positive transfectant of BW5147 (3B2) with a long term CTL line, 2C, and with CTLs generated in a mixed leucocyte reaction (MLR) shut off the CTL's Lyt-2 gene as expected but express the CTL's Lyt-3 gene product as a heterodimer with the product of the transfected Lyt-2 gene. Thus the Lyt-3 gene is not subject to the same negative regulatory influences as the Lyt-2 gene. That expression of Lyt-2 is not necessary for Lyt-3 gene transcription to continue is demonstrated by the finding that hybrids of MLR-generated CTLs with either BW5147 (Lyt-2-negative) or 3B2 (Lyt-2-positive) cells express Lyt-3 RNA. Southern hybridization and structural analysis of DNA fragments generated using the polymerase chain reaction demonstrated that hybrids contain several species of Lyt-3 RNA, one of which lacks the exon encoding the extracellular V-like domain and appears to be the product of an alternatively-spliced RNA transcript. PMID- 8423053 TI - HLA-B51 transgenic mice as recipients for production of polymorphic HLA-A, B specific antibodies. PMID- 8423054 TI - Polymorphism in the upstream regulatory regions of HLA-DRB genes. PMID- 8423055 TI - Allelic variation in the DQ subregion of the canine major histocompatibility complex: II. DQB. PMID- 8423056 TI - Regional localization of the nu mutation on mouse chromosome 11. PMID- 8423057 TI - Reactive glia as rivals in regulating neuronal survival. AB - Reactive gliosis is a response noted after nearly every type of CNS injury and involves both activated microglia and astroglia. Although many investigators believe that reactive glia in some way regulate the survival of injured neurons, the influence of glial elements upon damaged neural tissues remains uncertain. To examine relationships between reactive glia and neurons, secretion products from both microglia and astroglia are tested for their effects upon the survival of cultured neurons. Microglia are found to secrete neurotoxic agents, while astroglia are a source of neuronotrophic factors. Similar patterns of soluble factor production are noted for astroglia-rich or microglia-rich regions of rat neocortex damaged by ischemia. These observations suggest that microglia and astroglia compete for control of neuronal survival. Importantly, microglial neurotoxins might hinder the recovery of neurologic function at sites of inflammation. PMID- 8423058 TI - Cytotoxicity of microglia. AB - The most characteristic property of microglia is their swift activation in response to neuronal stress and their capacity for site-directed phagocytosis. The transformation of microglia into intrinsic brain macrophages appears to be under strict control and takes place if neuronal and/or terminal degeneration occurs in response to nerve lesion. The differentiation of microglia into brain macrophages is accompanied by the release of several secretory products, e.g., proteinases, cytokines, reactive oxygen intermediates, and reactive nitrogen intermediates. Interference with the microglial activation or the productions of cytotoxic metabolites by microglia may thus offer new therapeutic opportunities for the prevention of neuronal cell death in CNS disease. PMID- 8423059 TI - Microglia progenitor cells: a subpopulation in cultures of mouse neopallial astroglia. AB - We have shown that astroglia cultures initiated from newborn mouse neopallium contain microglia progenitor cells. Astroglia secrete growth factor(s) that transform the progenitor cells into microglia. One of the trophic factors is bone marrow macrophage growth factor CSF-1. Mouse embryonic fibroblast cells STO also secrete trophic factor(s) that acts synergistically or additively with CSF-1 on microglia progenitor cells. Using limiting dilution analysis we estimated that in the presence of CSF-1 and STO cells, 1 in every 8 cells in astroglia cultures is a potential microglia progenitor cell. PMID- 8423060 TI - Characterization of microglia and macrophages in the central nervous system of rats: definition of the differential expression of molecules using standard and novel monoclonal antibodies in normal CNS and in four models of parenchymal reaction. AB - This report describes the development of a new panel of monoclonal antibodies produced following immunization of mice with cultured rat microglial cells. Using these new reagents and previously defined antibodies that bind to microglia or macrophages, the responses of parenchymal microglia, perivascular "microglial" cells, and infiltrating macrophage/monocytes were examined in 4 divergent models of central nervous system reaction. These were brain abscess, experimental allergic encephalomyelitis, Wallerian degeneration, and stab wound. No single new antibody was specific only for microglia; all antibodies positively staining microglial cells also labeled various subsets of macrophage/monocytic cells in normal tissues of the immune system. Moreover, the results indicate that microglia are capable of different levels and a variety of types of response, as defined by the molecules they elaborate. These findings suggest that these CNS resident cells belong to the extended monocyte/macrophage/dendritic cell family and that they do not respond in a stereotypic manner to all forms of CNS insult. PMID- 8423061 TI - Microglial MHC antigen expression after ischemic and kainic acid lesions of the adult rat hippocampus. AB - By taking advantage of the specific neuronal and connective organization of the hippocampus and the different susceptibility of hippocampal neurons to transient cerebral ischemia or intraventricular injections of kainic acid (KA), we examined the microglial reactions to different types of neuronal injury. In all areas with neuronal or axonal degeneration, the microglial cells reacted by specific degeneration-related morphological transformations and expression of class I major histocompatibility complex (MHC) antigen. Subpopulations of microglial cells also expressed class II MHC antigen and leukocyte common antigen (LCA) in relation to (1) degenerating nerve cell bodies in the dentate hilus and the CA1 and CA3 pyramidal cell layers, (2) postischemic degeneration of dendrites in the stratum radiatum of CA1, and (3) combined dendritic and axonal degeneration in the stratum radiatum of the KA-lesioned CA3. MHC II and LCA expression was not observed in relation to degeneration of the CA3-derived Schaffer collaterals in CA1 after KA-induced CA3 lesions. In the case of ischemia the degeneration related reactions were preceded by an early, generalized microglial reaction, which also included areas without subsequent signs of neural degeneration. This reaction, which was transient and characterized by subtle morphological changes and induction of class I MHC antigen only, was presumably triggered by a general postischemic perturbation of the cerebral microenvironment, and not by actual neural degeneration. In conclusion, we found that microglial expression of class I MHC antigen was a sensitive marker of both the general perturbation after ischemia and axonal degeneration distant from the areas of actual nerve cell death.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423062 TI - A chronicle of microglial ontogeny. PMID- 8423063 TI - Altered antigen expression of microglia in the aged rodent CNS. AB - Microglia, the resident macrophages of the central nervous system, are characterised by a highly specialized morphology and unusual antigenic phenotype. Microglia appear to be downregulated by their microenvironment when compared to other tissue macrophages. We have studied the microglia in brains of healthy, aged rats with a panel of monoclonal antibodies. We have found that microglia in the brains of these aged rats express antigens that are downregulated or absent from microglia of juvenile rats. The stimuli which give rise to this upregulated phenotype are not known. Age related changes in the phenotype of microglia should be taken into account when considering the possible role of microglia in neuropathological conditions. PMID- 8423064 TI - Heterogeneity of microglial and perivascular cell populations: insights gained from the facial nucleus paradigm. AB - We reflect here on the development of a neuroimmunological concept which has been formulated over the past 5 years through studying microglial cell responses in the facial nerve system. A simple axotomy of the adult rat facial nerve which causes regeneration of facial motor neurons and little, if any, cell death can activate microglial cells just as easily as a full-blown degeneration of the entire nucleus induced by toxic ricin. In both instances, the prompt microglial reaction is characterized by a series of structural and phenotypic changes which are in many ways similar to an immune response, e.g., there is cell proliferation and upregulation of MHC antigens. However, since white blood cells do not participate in the retrograde response of facial motor neurons, we have adopted a notion which views microglia as a CNS-wide network of immunocompetent cells whose morphological dissimilarities from leukocytes are a result of their unique adaptation to the CNS architecture. We have continued our in vivo investigations of the phagocytic and immunophenotypic properties of microglial and perivascular cells during the retrograde reaction of facial motor neurons by using intra neural injections of fluorogold (FG) and ricin followed by lectin and immunostaining for microglia. Two new findings can be added to the microglial neuroimmune network: (1) Microglia take up FG only after motor neuron degeneration, whereas perivascular cells may take up FG under nondegenerating conditions. (2) Immunologically important molecules, such as MHC class II, CD4, and leukocyte common antigens, are expressed by different microglial subpopulations. Thus there is functional and phenotypic heterogeneity among immunocompetent cells of the CNS. PMID- 8423065 TI - Microglia and cytokines in neurological disease, with special reference to AIDS and Alzheimer's disease. AB - Microglia are associated with central nervous system (CNS) pathology of both Alzheimer's disease (AD) and the acquired immunodeficiency syndrome (AIDS). In AD, microglia, especially those associated with amyloid deposits, have a phenotype that is consistent with a state of activation, including immunoreactivity with antibodies to class II major histocompatibility antigens and to inflammatory cytokines (interleukin-1-beta and tumor necrosis factor alpha). Evidence from other studies in rodents indicate that microglia can be activated by neuronal degeneration. These results suggest that microglial activation in AD may be secondary to neurodegeneration and that, once activated, microglia may participate in a local inflammatory cascade that promotes tissue damage and contributes to amyloid formation. In AIDS, microglia are the primary target of retroviral infection. Both ramified and ameboid microglia, in addition to multinucleated giant cells, are infected by the human immunodeficiency virus (HIV-1). The mechanism of microglial infection is not known since microglia lack CD4, the HIV-1 receptor. Microglia display high affinity receptors for immunoglobulins, which makes antibody-mediated viral uptake a possible mechanism of infection. In AIDS, the extent of active viral infection and cytokine production may be critically dependent upon other factors, such as the presence of coinfecting agents. In the latter circumstance, very severe CNS pathology may emerge, including necrotizing lesions. In other circumstances, HIV infection of microglia probably leads to CNS pathology by indirect mechanisms, including release of viral proteins (gp120) and toxic cytokines. Such a mechanism is the best hypothesis for the pathogenesis of vacuolar myelopathy in adults and the diffuse gliosis that characterizes pediatric AIDS, in which very little viral antigen can be detected. PMID- 8423066 TI - Microglia in degenerative neurological disease. AB - Microglia express many leukocyte surface antigens which are upregulated in such chronic degenerative neurological diseases as Alzheimer's disease (AD) and amyotrophic lateral sclerosis (ALS). These surface antigens include leukocyte common antigen, immunoglobulin Fc receptors, MHC class I and class II glycoproteins, beta 2-integrins, and the vitronectin receptor. Ligands for these receptors are also found. They include immunoglobulins, complement proteins of the classical pathway, T lymphocytes of the cytotoxic/suppressor and helper/inducer classes, and vitronectin. T lymphocytes marginate along capillary venules, with some penetrating into the tissue matrix. Immunoglobulins and complement proteins are synthesized locally in brain, although they may also come from the bloodstream if the blood-brain barrier is compromised. The membrane attack complex, which is formed from C5b-9, the terminal components of complement, has been identified in AD and multiple sclerosis brain tissue. In addition, proteins designed to defend against bystander lysis caused by the membrane attack complex, including protectin, C8 binding protein, clusterin, and vitronectin, are associated with damaged neuronal processes in AD. Autodestruction may play a prominent part in these 2 diseases. PMID- 8423067 TI - The origin and nature of ramified and amoeboid microglia: a historical review and current concepts. AB - The origin of ramified microglia has been a longstanding controversial issue, with 4 major schools of thought, which state that they are derived (1) from invasion of mesodermal pial elements, (2) from neuroectodermal matrix cells together with the macroglia, (3) from pericytes, and (4) from invasion of monocytes in early development. This paper is in support of the last-mentioned hypothesis. It is known that ramified microglial cells do not divide under normal circumstances, and since our studies in the corpus callosum have shown that these cells do not appear until the fifth postnatal day, it is reasoned that they must be derived from some preexisting mitotically active cells. The putative precursor is the preponderant amoeboid microglia in the same region. Our experimental studies with the carbon labelling technique have demonstrated for the first time that blood monocytes invade into the early postnatal brain to become amoeboid microglia, which then differentiate into ramified microglia. Just like other tissue macrophages, the monocyte-derived amoeboid microglia exhibit features indicative of phagocytic activities. These include the content of hydrolytic enzymes, uptake of carbon, and a characteristic surface morphology, as seen by scanning electron microscopy. The transformation of amoeboid microglia into ramified microglia, which occurs between the second and third postnatal week, is considered to be a regressive phenomenon, as shown by the diminution of their content of hydrolytic enzymes and the downregulation of membrane antigen. Apart from their primary role as active phagocytes, their involvement in Alzheimer's disease (AD) is evidenced recently by the fact that the cells are specifically marked by antibodies present in the cerebrospinal fluid of AD patients. In conclusion, ramified microglial cells are derived from monocytes, but through an intermediate amoeboid microglia stage as active macrophages in the perinatal period. PMID- 8423068 TI - Reduction in oral immunogenicity of cholera toxin B subunit by N-terminal peptide addition. AB - The mucosal adjuvanticity of cholera toxin and the potential of the B subunit of cholera toxin (CtxB) to serve as an oral vaccine carrier have prompted interest in the coupling of immunogenic peptides to this protein. The purpose of this study was to determine how such fusions affect the function of CtxB. Oligonucleotides were genetically fused to the 5' terminus of the ctxB gene to encode additional amino acids of 8, 12, and 24 residues in length. None of these additions affected the ability of CtxB to oligomerize, as determined by nondenaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Circular dichroism revealed no difference in conformation between the modified B subunits, regardless of the length of the addition. However, when compared with native CtxB, additions to the N terminus induced a consistent change in the net conformation of the protein. By using a competitive enzyme immunoassay, the affinity of the modified B subunits for GM1 ganglioside was shown to gradually decrease with increasing length of the N-terminal addition. A similar pattern was observed for the ability of the chimeras to inhibit proliferation of concanavalin A-stimulated spleen cells in vitro, which is a previously described functional property of CtxB that is dependent on its binding to cells. Lastly, the oral immunogenicity of these chimeras was found to be less than that of native CtxB. These results indicate that large fusions to the N terminus of CtxB can significantly affect its biological properties and could reduce its value as a mechanism for effective mucosal immunization. PMID- 8423069 TI - Immunity and responses of circulating leukocytes and lymphocytes in monkeys to aerosolized staphylococcal enterotoxin B. AB - Rhesus monkeys immunized intramuscularly or orally with staphylococcal enterotoxin B (SEB) toxoid or SEB toxoid incorporated in microspheres made of poly(DL-lactide-co-glycolide) were challenged with a lethal dose of aerosolized SEB to study their immunity and cellular responses in the circulation. It was found that circulating antibodies play a critical role in preventing SEB from triggering toxicosis. Monkeys with high levels of antibodies survived, while those with low levels underwent 2 to 3 days of toxicosis and died. Intramuscular immunization induced high levels and oral immunization induced low levels of antibodies. The circulating antibodies in surviving monkeys decreased dramatically within 20 min and started to rebound at 90 min after SEB challenge. At 90 min, the dying monkeys showed in the circulation a dramatic increase of polymorphonuclear leukocytes and decreases of NK cells and monocytes (CD16 and CD56 markers) as well as of lymphocytes with HLA-DR, CD2, CD8, and IL2R alpha (CD25) markers. The number of polymorphonuclear leukocytes showed an inverse correlation with the numbers of monocytes and various lymphocyte subpopulations which, except for IL-2R, CD16, and CD56(+) cells, showed a direct correlation with one another. The changes in the populations of leukocytes, monocytes, NK cells, and lymphocytes seem to be an indication of initial toxicosis; however, the roles of these cells in toxicosis and death remain to be defined. PMID- 8423070 TI - Cell-mediated immunity to Bordetella pertussis: role of Th1 cells in bacterial clearance in a murine respiratory infection model. AB - A murine respiratory infection model was used to study the mechanism of protective immunity to Bordetella pertussis. We found that nude mice, which are deficient in T cells, developed a persistent infection and failed to clear the bacteria after aerosol inoculation. In contrast, normal adult nonimmune mice cleared a respiratory infection approximately 35 days after challenge. Before bacterial clearance, antipertussis antibody levels in serum were low or undetectable, whereas consistent antigen-specific T-cell responses were demonstrated throughout the course of infection. The in vitro responses detected in immune spleen cells were mediated by a population of CD4+ major histocompatibility complex class II-restricted Th1-like cells that secreted interleukin-2 and gamma interferon but not interleukin-4. Adoptive transfer of immune spleen cells into nude or sublethally irradiated immunosuppressed mice before challenge resulted in bacterial clearance within 14 to 21 days. In contrast, injection of serum from convalescent mice before challenge only marginally reduced the bacterial load early in the course of infection. Furthermore, transfer of enriched T cells or purified CD4+ T cells but not CD8+ T cells from immune mice conferred a high level of protection. Recipients of CD4+ T cells cleared the bacteria from the lungs within 20 days of challenge, at which time B. pertussis-specific antibodies in the serum were undetectable. Although we do not rule out a contribution of mucosal immunoglobulin A, our findings suggest that cellular responses mediated by CD4+ Th1 cells play an important role in protective immunity to B. pertussis. PMID- 8423071 TI - Cloning, heterologous expression, and characterization of the Erysipelothrix rhusiopathiae DnaK protein. AB - The dnaK (hsp70) gene from the facultative intracellular pathogen Erysipelothrix rhusiopathiae was cloned by heterologous DNA hybridization of a genomic library using the Escherichia coli dnaK gene as a probe. A 3.2-kb fragment which encoded an 1,800-bp open reading frame was recovered. The deduced amino acid sequence of this open reading frame shares 56% identity with the E. coli DnaK protein. Expression of the encoded protein in E. coli by using the phage T7 promoter/polymerase system resulted in accumulation of a unique 65-kDa protein. Western blot (immunoblot) analysis of extracts from a recombinant E. coli strain using anti-E. coli DnaK polyclonal antibodies confirmed that the cloned gene encodes a DnaK homolog. The recombinant E. rhusiopathiae DnaK protein was purified to 80% homogeneity by ATP affinity chromatography. The purified material hydrolyzed ATP with a specific activity of 100 nmol min-1 mg of protein-1. Analysis of total protein extracts from E. rhusiopathiae indicates that DnaK is a highly expressed protein in this organism. PMID- 8423072 TI - Mycobacterial antigen complex A60-specific T-cell repertoire during the course of pulmonary tuberculosis. AB - The Mycobacterium bovis antigen complex A60 is known to be immunodominant in tuberculosis and to have a protective effect against experimental infection in vitro and in vivo. To identify immunodominant and possibly protective antigens in pulmonary tuberculosis, the T-cell repertoire directed to nitrocellulose-bound fractions of A60 antigen was analyzed in active tuberculosis patients during the course of the infection and after recovery. The results show that patients infected with Mycobacterium tuberculosis acquired complete A60-T-cell reactivity only in the late phases of infection. At disease onset, patients with active tuberculosis were characterized by (i) T-cell unresponsiveness to most A60 fractions, (ii) high tumor necrosis factor alpha production, and (iii) low gamma interferon (IFN-gamma) release. Several weeks after chemotherapy, the unresponsive state disappeared and the following reverse situation was observed: (i) high blastogenic response to almost all A60 fractions, (ii) low tumor necrosis factor alpha release, and (iii) high IFN-gamma production. In addition, 60% of these patients significantly responded against seven A60 fractions (61 to 58, 56 to 53, 49 to 46, 46 to 44, 35 to 33, 33 to 30, and 30 to 28 kDa), indicating that they included immunodominant antigens. Furthermore, only the fractions within the molecular mass ranges of 56 to 44 and 35 to 28 kDa induced IFN-gamma synthesis. One year after complete recovery from infection, more than 60% of past-active tuberculosis subjects had memory T cells specific for the immunodominant fractions of 61 to 58, 56 to 53, 49 to 46, and 33 to 30 kDa. Since the same fractions induced the strongest IFN-gamma production, known to exhibit antimycobacterial effects, it is suggested that these may represent the inducers of a protective immune response. PMID- 8423073 TI - Comparison of the alpha-toxin genes of Clostridium perfringens type A and C strains: evidence for extragenic regulation of transcription. AB - The Clostridium perfringens plc gene encoding phospholipase C (alpha-toxin) was cloned from type C NCIB 10662, a strain which produces low levels of phospholipase C activity. The nucleotide sequence of a cloned 3.1-kb HindIII fragment was determined. The same fragment was also cloned from type A NCTC 8237, a phospholipase C-overproducing strain. In this case, an open reading frame (ORF2) truncated in the previously cloned 2-kb fragment was also sequenced. Comparison of the nucleotide sequence between the 3.1-kb fragments of the two type strains shows some differences both in the plc gene and in ORF2. However, when the 3.1-kb fragment was cloned into plasmid pUC19 and expressed in Escherichia coli, the plc genes from both type strains were similarly expressed and the toxins produced showed similar levels of activity. Northern blot analysis revealed that the type A strain produced 16 to 23 times more plc mRNA than the type C strain. These results indicate that in C. perfringens the two plc genes are transcribed at different rates, probably because of a difference in a locus lying outside of the cloned fragments. Gel retardation analysis showed that the type A strain possessed two different proteins that bound different regions of the plc gene. However, one of these proteins, which binds within the plc coding region, was not found in the type C strain, suggesting that it plays a role in the regulation of the plc gene expression. PMID- 8423074 TI - Effect of polysaccharide capsule and methods of preparation on human lymphocyte proliferation in response to Cryptococcus neoformans. AB - Cryptococcus neoformans is a pathogenic encapsulated yeast that infects patients that have defective cell-mediated immunity, including AIDS patients. Whole cryptococcal organisms that are killed by heating stimulate normal human lymphocytes to proliferate. However, strains of C. neoformans vary widely in virulence and therefore in their ability to cause disease in humans. To determine the effect of virulence factors such as the cryptococcal capsule, serotype, and the state of the organisms on the lymphocyte response to C. neoformans, human peripheral blood mononuclear cells were stimulated with C. neoformans in vitro and lymphocyte proliferation was determined. The major determinant of the lymphocyte response to C. neoformans was the amount of polysaccharide present. The response was greater after stimulation by minimally encapsulated strains (strains C3D, 68, and 613) than by heavily encapsulated strains (strains 6 and 145). A heavily encapsulated strain (strain 6) did not suppress the response to an acapsular mutant (strain 67). However, the response to an acapsular strain was suppressed by the addition of purified polysaccharide. Human lymphocytes responded to both serotypes of C. neoformans var. neoformans. The antigen responsible for lymphocyte stimulation was preserved despite various techniques of inactivation, including heat, paraformaldehyde fixation, irradiation, and mechanical disruption. Finally, lymphocytes responded equally to live and killed organisms. These results suggest that capsular polysaccharide, a known virulence factor, may suppress the human lymphocyte response to C. neoformans during an infection. Lymphocytes could respond to C. neoformans regardless of the viability of the organism, and they could also respond to disrupted organisms. We speculate that lymphocyte proliferation in vitro could be related to the protective immune response in host defense to C. neoformans and that it is suppressed by virulence factors of C. neoformans. PMID- 8423075 TI - Cloning and sequencing of Coxiella burnetii outer membrane protein gene com1. AB - The gene for an approximately 27-kDa outer membrane-associated, immunoreactive protein was cloned from the rickettsial pathogen Coxiella burnetii. The gene, designated com1 for Coxiella outer membrane protein 1, was expressed in Escherichia coli, presumably by its own promoter. The complete nucleotide sequence of the gene was determined. The deduced amino acid sequence of 252 residues includes a putative leader sequence. The leader sequence is recognized in and removed by E. coli on the basis of the difference in the molecular mass of the protein produced in an in vitro transcription-translation system (27.6 kDa) and that of the protein immunoprecipitated from an iodinated E. coli clone (25.7 kDa). The Com1 protein expressed in E. coli was proteinase K sensitive in nondisrupted cells and soluble in 1% Sarkosyl, suggesting a loose association with the outer membrane. While the complete predicted sequence of the Com1 protein does not show any overall similarity to those of previously described proteins, a region which includes the only two cysteines in Com1 is homologous to the catalytic site of protein disulfide oxidoreductases. PMID- 8423076 TI - Group B streptococci invade endothelial cells: type III capsular polysaccharide attenuates invasion. AB - Group B streptococci (GBS) are the most common cause of neonatal sepsis and pneumonia. The pathogenesis of GBS disease is not completely defined. GBS-induced endothelial cell injury is suggested by histological findings at autopsy and in animal studies. We hypothesized that (i) type III GBS (COH-1) invade and injure human umbilical vein endothelial (HUVE) cells and (ii) isogenic mutations in GBS capsule synthesis would influence HUVE invasion. Confluent HUVE monolayers were infected for 0.5, 2, or 6 h. Media with penicillin plus gentamicin were added and incubated for 2 h to kill extracellular bacteria. Cells were washed and lysed, and the number of live intracellular bacteria was determined by plate counting. COH-1 invaded HUVE cells in a time-dependent manner at levels 1,000-fold higher than those of the noninvasive Escherichia coli strain but significantly lower than those of Staphylococcus aureus. There was no evidence for net intracellular replication of GBS within HUVE cells. COH-1 infection of HUVE cells caused the release of lactate dehydrogenase activity. GBS invasion was inhibited by cytochalasin D in a dose-dependent manner; GBS-induced lactate dehydrogenase release was attenuated by cytochalasin D. The isogenic strains COH 1-11, devoid of capsular sialic acid, and COH 1-13, devoid of all type III capsule, invaded HUVE cells three- to fivefold more than the parent COH-1 strain. The type III capsular polysaccharide and particularly the capsular sialic acid attenuate GBS invasion of HUVE cells. Electron micrographs of lung tissue from a GBS-infected newborn Macaca nemestrina also showed GBS within capillary endothelial cells. We conclude that endothelial cell invasion and injury are potential mechanisms in the pathogenesis of GBS disease. PMID- 8423077 TI - Bordetella pertussis and Bordetella parapertussis: two immunologically distinct species. AB - Bordetella pertussis and Bordetella parapertussis are closely related species. Both are responsible for outbreaks of whooping cough in humans and produce similar virulence factors, with the exception of pertussis toxin, specific to B. pertussis. Current pertussis whole-cell vaccine will soon be replaced by acellular vaccines containing major adhesins (filamentous hemagglutinin and pertactin) and major toxin (pertussis toxin). All of these factors are antigens that stimulate a protective immune response in the murine respiratory model and in clinical assays. In the present study, we examined the protective efficacies of these factors, and that of adenylate cyclase-hemolysin, another B. pertussis toxin, against B. parapertussis infection in a murine respiratory model. As expected, pertussis toxin did not protect against B. parapertussis infection, since this bacterium did not express this protein, but the surprising result was that none of the other factors were protective against B. parapertussis infection. Furthermore, B. parapertussis adenylate cyclase-hemolysin, although it protected against B. parapertussis infection, did not protect against B. pertussis infection. Despite a high degree of homology between both B. pertussis and B. parapertussis species, no cross-protection was observed. Our results outline the fact that, as in other gram-negative bacteria, Bordetella surface proteins vary immunologically. PMID- 8423078 TI - Gamma interferon-induced nitric oxide production reduces Chlamydia trachomatis infectivity in McCoy cells. AB - McCoy cells, murine-derived cells commonly used for propagation of chlamydiae, were found to be efficient producers of nitric oxide (NO) when primed with murine gamma interferon (IFN-gamma) and then exposed to the second signals provided by Escherichia coli lipopolysaccharide, human interleukin-1 alpha, murine tumor necrosis factor alpha, or Chlamydia trachomatis type H. Murine recombinant IFN gamma over a range of 0 to 50 U/ml inhibited infectivity of C. trachomatis type H in a dose-dependent fashion in McCoy cells while simultaneously inducing NO production. Quantitation of infectious chlamydia progeny remaining in McCoy cells 48 or 72 h postinfection revealed that IFN-gamma-primed McCoy cells reduced chlamydial inclusion-forming units (expressed as units per milliliter) by 4 log10 units at higher IFN-gamma concentrations (50 U/ml) compared with control values. The magnitude of this antichlamydial effect was directly related to increased synthesis of NO, the production of which was IFN-gamma dose dependent. The antichlamydial effects of IFN-gamma were blocked in a dose-dependent manner by the addition of N-guanidino-monomethyl L-arginine (MLA), an inhibitor of nitric oxide synthesis. These results suggest that although IFN-gamma priming of McCoy cells is required for antichlamydial activity, nitric oxide is a necessary effector molecule involved in the mechanism(s) of IFN-gamma-induced inhibition of chlamydial proliferation in this murine cell line. The ability to block the potent antichlamydial effects of IFN-gamma by inhibition of a specific enzyme, nitric oxide synthase, may give insights into mechanisms by which IFN-gamma and perhaps other cytokines are able to control proliferation of chlamydiae and other intracellular pathogens. PMID- 8423079 TI - Effects of antibody isotype and host cell type on in vitro neutralization of Chlamydia trachomatis. AB - Monoclonal antibodies (MAbs) E-4, E-21, and DIII A3, which recognize the same or similar overlapping peptides in the variable domain IV of the major outer membrane protein of Chlamydia trachomatis but differ in isotype, were used in a complement-independent (CI) in vitro neutralization assay. These MAbs had previously been shown to neutralize chlamydial infectivity in HeLa 229 cells in a complement-dependent assay. In this report, all three MAbs neutralized chlamydial infectivity in HaK cells in a CI assay. However, when HeLa cells were used as the host cell, MAb E-4 (immunoglobulin G2b [IgG2b]) and MAb DIII A3 (IgG2b) failed to neutralize infectivity, while MAb E-21 (IgG1) neutralized chlamydial infectivity. These findings are consistent with the proposal that because of the presence of Fc gamma RIII receptors, HeLa cells facilitate infectivity and thus block neutralization through the uptake of an IgG2b-chlamydia complex. Since Fc gamma RIII receptors do not bind or bind poorly to IgG1, neutralization of C. trachomatis by MAb E-21 in HeLa cells is also corroborative evidence for the role of Fc gamma RIII receptors in this interaction. A fivefold enhancement of infectivity was seen when 10 and 1 micrograms of MAb E-4 per ml were tested in a CI assay with HeLa cells. In performing CI neutralization synergy studies in HeLa cells with MAbs E-4 and E-21, antagonism between MAbs E-4 and E-21 was observed at MAb E-4 concentrations of 10 and 1 micrograms/ml for all concentrations of MAb E-21 tested (10 to 0.1 micrograms/ml). When HaK cells were used in the same studies, no antagonism between the MAbs was found. In addition, when HeLa cells were used in a CI assay, polyclonal serum raised to a peptide representing variable domain IV of the major outer membrane protein inhibited the neutralizing ability of MAb E-21. The blocking of neutralization and the enhancement of infectivity by chlamydia-specific antibodies seen in this investigation with HeLa cells may have important clinical implications for developing preventive strategies for chlamydial infections. PMID- 8423080 TI - The Salmonella typhimurium virulence plasmid increases the growth rate of salmonellae in mice. AB - The virulence plasmids of Salmonella typhimurium and other invasive Salmonella serovars have long been associated with the ability of these bacteria to cause systemic infection beyond the intestines in orally inoculated animals. Genetic analysis of virulence genes on the high-molecular-weight plasmids has revealed that no more than five genes spanning a 6.2-kb region are sufficient to replace the entire plasmid for conferring virulence. However, the exact virulence function(s) encoded by these genes has not been elucidated. In this report, we measured the possible effect of the virulence plasmid on the growth rate of S. typhimurium in mice by two complementary procedures. The first procedure used segregation of a temperature-sensitive plasmid in vivo to provide a measure of bacterial divisions and the number of recovered marker plasmid-containing salmonellae as a measure of killing. In the second procedure, aroA deletions were transduced into virulence plasmid-containing and plasmid-cured S. typhimurium. Since AroA- salmonellae are inhibited for growth in vivo, if the virulence plasmid affected only growth rate, no difference in the recoveries of the paired AroA- strains would be seen. Virulence plasmid-containing S. typhimurium segregated the marker plasmid more rapidly than did the virulence plasmid-cured strain, and AroA- derivatives of both strains were recovered equally from mice. Therefore, the S. typhimurium virulence plasmid increased growth rate but had no detectable effect on killing or bacterial movement into deep tissues. To examine whether the plasmid accomplished this function by affecting the intracellular/extracellular location of bacteria, orally infected mice were injected with gentamicin to kill the extracellular bacteria. Wild-type and plasmid-cured S. typhimurium strains were equally resistant to gentamicin in vivo and hence most likely located intracellularly to equal degrees. When wild-type and plasmid-cured S. typhimurium strains were sequestered within peritoneal chambers in mice, the resulting extracellular growth was equal. Therefore, the virulence plasmid increases the growth rate of S. typhimurium in mice, probably within mouse cells. PMID- 8423081 TI - Antibiotics enhance binding by human lipid A-reactive monoclonal antibody HA-1A to smooth gram-negative bacteria. AB - The effect of antibiotic exposure of phenotypically smooth gram-negative bacteria on binding by the human lipid A-reactive monoclonal antibody HA-1A (trademark of Centocor, Inc.) was examined by liquid-phase immunoassay and by dual-parameter flow cytometry (fluorescence-activated cell sorter [FACS]) analysis. HA-1A exhibited dose-dependent binding to untreated rough gram-negative bacteria such as the Escherichia coli D21F2 Re chemotype strain but little binding to untreated smooth strains such as E. coli O111:B4, or to gram-positive bacteria. However, overnight incubation of E. coli O111:B4 with inhibitory concentrations of ceftazidime produced dose-dependent enhancement of HA-1A binding. Similar augmentation of HA-1A binding was observed when other smooth strains were exposed to cell wall-active agents. Dual-parameter FACS analysis of E. coli O111:B4 exposed overnight to two times the MIC of ceftazidime revealed a decrease in forward light scatter, indicating a reduction in average cell size or bacterial fragmentation, accompanied by a striking increase in lipid A-inhibitable HA-1A binding. Moreover, ceftriaxone, but not gentamicin, produced a marked increase in propidium iodide uptake, indicating an increase in bacterial cell permeability, and a corresponding enhancement of HA-1A binding. Antibiotic-induced enhancement of HA-1A binding to smooth strains of gram-negative bacteria thus appears related to specific alterations in bacterial cell morphology resulting in exposure of the epitope recognized by HA-1A. PMID- 8423082 TI - Cysteine-dependent zinc binding by membrane proteins of Giardia lamblia. AB - The abundant, highly variable surface proteins (VSPs) which cover the surface of Giardia lamblia trophozoites compose a group of extremely cysteine (C)-rich proteins in which more than half of the cysteines are in the motif CXXC. Because of the constancy of these features among the known VSPs and the prominence of cysteine and particularly CXXC in proteins that bind zinc and other metals, we asked whether G. lamblia VSPs bind zinc in vitro. VSPs are the major protein component of Triton X-114 detergent-phase extracts of G. lamblia trophozoites and can be readily identified by surface iodination of intact cells. The partitioning of 65Zn binding into the Triton X-114 detergent phase and the correspondence between surface iodination and zinc binding patterns of four G. lamblia strains or sublines with different VSPs support the idea that VSPs bind zinc. The requirement for renaturation of blots with a reducing agent indicates that Zn2+ is coordinated by cysteine residues, rather than by other amino acids. Binding did not appear to be specific to zinc since it was inhibited by competition with other divalent metal ions. The abundance of the VSPs and the prevalence of metal binding motifs among all known variants suggest that they may play an important role in trophozoite survival and colonization in the host. PMID- 8423083 TI - Suppression of T-cell proliferation by CD8+ T cells induced in the presence of protoscolices of Echinococcus multilocularis in vitro. AB - Immunoregulatory influences of protoscolices (PSC) of Echinococcus multilocularis on murine T-lymphocyte functions have been examined in an in vitro system. Proliferative responses of spleen cells stimulated with concanavalin A (ConA) or anti-CD3 monoclonal antibodies were depressed by the addition of PSC. In the presence of PSC, both interleukin-2 (IL-2) production and IL-2 receptor (IL-2R) expression by lymphocytes stimulated with ConA were significantly reduced. However, exogenous IL-2 reconstituted both the ConA-stimulated proliferative responses and IL-2R expression. These findings suggest that PSC of E. multilocularis can suppress lymphoid cell responses via influences on IL-2 production. Indeed, addition of CD(8+)-enriched cells from cultures stimulated with ConA plus PSC to fresh spleen cells showed marked suppression of the ConA responses. IL-2 production as well as IL-2R expression on the spleen cells so treated were suppressed. These findings reveal a suppressive immunologic function induced by E. multilocularis PSC that involves inhibition of IL-2 production and reduction of IL-2R expression. The PSC-induced CD8+ cells appear to play a key role in the suppressive regulation of host immune responses against E. multilocularis. PMID- 8423084 TI - Shiga-like toxin II-related cytotoxins in Citrobacter freundii strains from humans and beef samples. AB - By hybridizing colonies grown from 928 individual stool samples of patients suffering from diarrhea with oligonucleotide probes 772 and 849 complementary to Shiga-like toxin I (SLT-I) and SLT-II gene sequences, respectively, we identified two strains that hybridized with probe 849, which biochemical identification revealed as Citrobacter freundii. An additional five slt-II probe-positive isolates were screened from 81 beef samples. Polymerase chain reaction analysis and restriction of amplified products provided evidence for slt-II-related genes in all seven strains. From C. freundii LM 76, the genes encoding the A and B subunits were cloned in pUC 18 vectors and sequenced. The gene encoding the A subunit differed from that of Escherichia coli slt-IIvhc in 4 bases, resulting in two amino acid residue differences. In 11, 13, and 11 nucleotides, differentiation of slt-IIA, slt-IIcA, and vtx2haA, respectively, was found. These differences affected the predicted amino acid sequence as follows: there were six amino acid differences with SLT-IIA, five with SLT-IIcA, and four with VTx2haA. The nucleotide sequence of the gene encoding the B subunit is identical to slt IIvhcB and differed from slt-IIcB and vtx2haB by only a single nucleotide base, but this resulted in a predicted amino acid sequence identical to that reported for these toxins. We therefore termed the toxin genes C. freundii slt-IIcA and slt-IIcB. Culture filtrates inoculated with material from the colonies from primary cultures were cytotoxic to Vero cells. Neutralization assays with antisera to E. coli SLT-I, SLT-II, and SLT-IIvhc revealed that antibodies against SLT-IIvhc reduced the C. freundii cytotoxic activity specifically and to the same degree as with the E. coli SLT-IIvhc control strain. In five of the seven strains tested, subcultivation on both a liquid or solid medium resulted in loss of cytotoxic activity. With polymerase chain reaction, we demonstrated that loss of cytotoxic activity ran parallel with the loss of slt genes. These data demonstrate the intergeneric occurrence of SLT-II-related toxins, which may well be a new marker of enteropathogenicity in C. freundii. Our findings that the toxin genes belong to the slt-II family plus their evident instability in the majority of strains should help pave the way to a better understanding of their role in diarrhea or food poisoning. PMID- 8423085 TI - Suppression of lymphocyte and neutrophil functions by Pseudomonas aeruginosa mucoid exopolysaccharide (alginate): reversal by physicochemical, alginase, and specific monoclonal antibody treatments. AB - The mucoid exopolysaccharide (MEP or alginate) of Pseudomonas aeruginosa is thought to be a virulence factor for this organism by virtue of its ability to suppress local host defense mechanisms. We purified MEP from clinical isolates of mucoid P. aeruginosa, subjected it to degradation by ultrasonication, heat, alkali, and alginase, and reacted it with monoclonal antibodies specific for MEP epitopes. Partial reversal or complete abrogation of the inhibitory effects of alginate on human neutrophil random migration, chemotaxis, and hexose monophosphate shunt activity and lymphocyte transformation were observed following most of these treatments. Physicochemical analysis of degraded MEP revealed a positive correlation between changes in molecular size and viscosity and loss of biological properties. The biological properties of MEP were also shown to be dependent on the structural integrity of the O-acetyl groups substituted for the mannuronic acid residues. The results show that the capacity of MEP to suppress neutrophil and lymphocyte functions is dependent on its acetyl content and the physical properties of large size and viscosity and may provide part of the explanation for the propensity of mucoid P. aeruginosa to persist in the airways of patients with cystic fibrosis. These findings highlight the important role of MEP as one of the virulence factors in the pathogenesis of inflammatory damage and subsequent pulmonary destruction in cystic fibrosis. PMID- 8423086 TI - Molecular characterization of a protective outer membrane lipoprotein (OmlA) from Actinobacillus pleuropneumoniae serotype 1. AB - An expression library was constructed from an Actinobacillus pleuropneumoniae serotype 1 clinical isolate using a plasmid vector. The library was screened with serum raised against the culture supernatant of this strain. One Escherichia coli transformant which also reacted with convalescent serum was isolated and found to express a protein with an electrophoretic mobility of approximately 50,000. The A. pleuropneumoniae-derived DNA encoding the protein was localized and characterized by nucleotide sequence analysis and primer extension mapping. One open reading frame of 1,095 bases was detected and confirmed by TnphoA insertion mutagenesis. It encoded a protein with a calculated molecular mass of 40 kDa which was lipid modified and present in the outer membrane and in membrane blebs of A. pleuropneumoniae. This protein was designated as outer membrane lipoprotein A (OmlA), and the encoding gene as omlA. Southern blotting under low-stringency conditions revealed the presence of hybridizing sequences in all A. pleuropneumoniae type strains, and a specific serum detected a homologous protein in serotypes 2, 8, 9, 11, and 12 type strains. Pigs immunized with this recombinant protein preparation were protected from death in an aerosol challenge experiment with an A. pleuropneumoniae serotype 1 isolate. PMID- 8423087 TI - Antibody responses in the serum and respiratory tract of mice following oral vaccination with liposomes coated with filamentous hemagglutinin and pertussis toxoid. AB - Mice were orally vaccinated with liposomes coated with filamentous hemagglutinin (FHA) and detoxified pertussis toxin (PT) of Bordetella pertussis. FHA- and PT specific immunoglobulin G (IgG) was detected in serum, and both IgG and IgA were detected in lung washes following the immunization. Antibody responses in mice immunized with liposomes coated with FHA and PT were significantly higher than those in mice immunized with free FHA and PT, which demonstrated the adjuvanticity of the liposome carrier. The results indicate the potential usefulness of this approach for eliciting immune responses against FHA and PT (and perhaps other pertussis antigens) in humans and its possible utility in large-scale vaccination to protect against both B. pertussis infection and disease. PMID- 8423088 TI - Sequencing of leucocidin R from Staphylococcus aureus P83 suggests that staphylococcal leucocidins and gamma-hemolysin are members of a single, two component family of toxins. AB - A 2,813-bp HincII-ClaI DNA fragment encodes the two S and F components (LukS-R and LukF-R) of leucocidin R (Luk-R) which are secreted by Staphylococcus aureus P83. The two genes (lukS-R and lukF-R) belong to a single operon. Two peptidic sequences were deduced: LukS-R is a 35,721-Da polypeptide of 315 amino acids, including a signal sequence of 29 residues, and LukF-R is a 36,838-Da polypeptide of 325 amino acids, including a signal sequence of 25 residues. LukS-R and LukF-R were expressed in Escherichia coli and purified from the periplasmic space. Luk-R exerts biological activities on polymorphonuclear cells and on erythrocytes from various animals. Comparison of the amino acid sequence of LukF-R with that of the B component of gamma-hemolysin (HlgB), those of the F and S components of another recently sequenced staphylococcal leucocidin, and those of a few peptides of the F component from Panton-Valentine leucocidin suggests that all four toxins belong to a single, two-component family of toxins. PMID- 8423089 TI - Selective cytokine production by epithelial cells following exposure to Escherichia coli. AB - This study compared the repertoire of cytokines produced by epithelial cell lines and human peripheral blood monocytes in response to Escherichia coli. The A-498 and J82 urinary tract epithelial cell lines and human peripheral blood monocytes were exposed to E. coli Hu734. The cytokine content of single cells was detected by indirect immunofluorescence using monoclonal antibodies to interleukin-1 alpha (IL-1 alpha), IL-1 beta, tumor necrosis factor alpha (TNF-alpha), TNF-beta, IL-6, IL-8, and granulocyte macrophage-colony-stimulating factor, and the number of positive cells was used to quantitate the response. The J82 bladder cell line stained positive for IL-6, IL-8, and IL-1 alpha. The IL-8 and IL-6 response peaked at 2 h, while the number of IL-1 alpha-positive cells reached a peak 6 h after E. coli stimulation. The A-498 kidney cell line stained for IL-8 with a peak at 2 h and IL-6 with a peak at 6 h after E. coli stimulation. Peripheral blood monocytes stained for the cytokines IL-1 alpha, IL-1 beta, IL-8, IL-6, and TNF-alpha but not for TNF-beta and granulocyte macrophage-colony-stimulating factor after stimulation with E. coli. The results demonstrated that bacteria activated a cytokine response in the epithelial cell lines and monocytes. The epithelial cell lines had a more limited cytokine response profile than circulating monocytes, which may serve to limit the consequences of microbial exposure at the mucosal surface and help maintain the integrity of other tissue compartments. PMID- 8423090 TI - Oral immunization of mice and swine with an attenuated Salmonella choleraesuis [delta cya-12 delta(crp-cdt)19] mutant containing a recombinant plasmid. AB - Salmonella choleraesuis chi 3781, an attenuated [delta cya-12 delta(crp-cdt)19] mutant, was electroporated with the plasmid pBA31-R7, which codes for the expression of a 31-kDa protein from Brucella abortus (BCSP31). Recombinant S. choleraesuis chi 3781 stably maintained the pBA31-R7 plasmid and continued to express the cloned protein following recovery of the organism from orally inoculated animals. Unlike previous studies using S. typhimurium chi 4064(pBA31 R7), S. choleraesuis chi 3781(pBA31-R7) was able to colonize both the gut mucosa and deep tissues of both BALB/cByJ mice and crossbred swine. Orally inoculated mice developed serum antibodies to both the cloned 31-kDa protein (rBCSP31) and to S. choleraesuis chi 3781 endotoxin. These mice also developed a local intestinal antibody response to Salmonella endotoxin but not to rBCSP31. Similarly, mice inoculated with recombinant S. choleraesuis chi 3781 did not develop a delayed-type hypersensitivity (DTH) footpad response following injection with rBCSP31; however, these mice did respond to S. choleraesuis chi 3781 soluble antigen. Conversely, orally inoculated swine did not develop significant serum or intestinal antibody responses to cloned protein or Salmonella endotoxin, but DTH responses to both cloned protein and S. choleraesuis chi 3781 soluble antigen were strongly positive. The cell-mediated nature of these DTH responses was confirmed by histological examination. Results suggest that S. choleraesuis chi 3781 may be a suitable choice for further studies of vaccine efficacy in swine, especially for diseases which require cell mediated immunity for resolution. PMID- 8423091 TI - Modulating effect of dietary carbohydrate supplementation on Candida albicans colonization and invasion in a neutropenic mouse model. AB - We studied the effect of dietary carbohydrate supplementation on Candida albicans colonization and invasion of the gastrointestinal tract in a neutropenic mouse model. Mice inoculated with C. albicans were allowed free access to standard chow and drinking water supplemented with either glucose or xylitol or no carbohydrates (control). On days 33 through 36 postinoculation, the mean +/- standard error log10 CFU of C. albicans per gram on the mucosal surface, determined by quantitating CFU dislodged in the first wash of the gastric wall, was significantly higher in mice given the glucose supplement: 7.20 +/- 0.09 (glucose) versus 5.38 +/- 0.28 (xylitol) and 5.11 +/- 0.33 (control) CFU/g (P < or = 0.05 for each comparison by Fisher's protected least-significant-difference test). Fecal cultures also yielded the highest quantities of C. albicans in the glucose group. Invasion of the gastric wall by C. albicans correlated well with surface colonization in glucose-supplemented animals. Eight of 10 mice in this group, all with > 10(6) CFU/g, showed extensive invasive growth, as compared with only 2 of 26 mice in the remaining groups (P = 0.00006 by Fisher's exact test). These results indicate that dietary glucose intake is a key determinant of C. albicans growth in the gastrointestinal tract. The data provide an experimental rationale for clinical trials to decrease the intake of glucose or its utilization by C. albicans in immunocompromised patients. PMID- 8423092 TI - Identification by two-dimensional gel electrophoresis of a 58-kilodalton tumor necrosis factor-inducing protein of Mycobacterium tuberculosis. AB - We have previously identified proteins in fractions of culture filtrate of Mycobacterium tuberculosis with the capacity to induce cytokine production in monocytes, by using a technique we defined as "monocyte Western blotting" (immunoblotting). In this series of experiments, we have extended this technique to two-dimensional gel electrophoresis and have identified a novel 58-kDa protein of M. tuberculosis which induces production of tumor necrosis factor by human monocytes. Nitrocellulose particles bearing this protein were used to develop murine monoclonal antibodies by the technique of intrasplenic immunization. The protein was purified by preparative isoelectric focusing and gel electrophoresis and subjected to N-terminal amino acid sequence analysis. As tumor necrosis factor is a mediator of both pulmonary necrosis and macrophage activation for intracellular killing, this 58-kDa protein may play an important role both in the immunopathogenesis of tuberculosis and in mycobacterial immunity. PMID- 8423093 TI - Impact of experimental genital mycoplasmosis on pregnancy outcome in Sprague Dawley rats. AB - Specific-pathogen-free (SPF) female Sprague-Dawley rats were infected by intravaginal inoculation with 3 x 10(7) CFU of Mycoplasma pulmonis X1048 in 0.1 ml of Frey's broth or with an equal volume of sterile Frey's broth. A minimum of 10 days postinfection, rats were bred to noninfected males. Rats were necropsied at days 11, 14, and 18 of gestation and within 24 h of parturition. Throughout pregnancy, at least 50% of rats remained infected in the lower genital tract. At parturition, the major site of colonization was the respiratory tract (P = 0.02). M. pulmonis was not isolated from any site of any control rat. Pregnancy outcome was adversely affected by infection with M. pulmonis. Infected rats had significantly smaller litter sizes at day 18 of gestation (P < or = 0.01) and at term (P < or = 0.004). No statistically significant differences among the gestational stages in infected rats were noted for litter size. Total litter weight is a reflection of individual pup weight and of the number of pups born. Therefore, it was obvious that infected rats would have a significantly lower (P < or = 0.008) total litter weight than noninfected controls. However, when individual pup weights were considered, infected pups (n = 49) also had significantly lower (P < or = 0.0001) birth weights than did noninfected controls (n = 68). The incidence of an adverse pregnancy outcome at term (stillbirths, macerated fetuses, or resorptions) was higher (P < or = 0.01) in infected rats than in noninfected control rats. No stillborn pups or macerated fetuses were observed in any control term rats (n = 5). All control rats had live-born pups. Three infected rats had no live-born offspring. Resorptions were more common in infected rats than in control rats (P < or = 0.01). The mean number of resorptions per rat was greater in rats which went to term than in rats necropsied during gestation, indicating that the severity of disease was progressive. The rat is frequently the laboratory animal of choice for a wide variety of reproductive studies, and the experimental parameters that are most often measured (litter size, pup weight, and neonatal survival) were all adversely affected by genital mycoplasmosis. Genital mycoplasmosis is important as an animal model for the interaction of infectious agents and the host during pregnancy as well as in its own right as a confounding variable affecting research projects which use the rat as a model to study reproductive function and physiology. PMID- 8423094 TI - Light emission from a Mudlux transcriptional fusion in Salmonella typhimurium is stimulated by hydrogen peroxide and by interaction with the mouse macrophage cell line J774.2. AB - Hydrogen peroxide is known to induce a multigenic response in Salmonella typhimurium cells. We have used a Mudlux transcriptional reporter system to identify and isolate fusions in the virulent strain SL1344 which respond to hydrogen peroxide in vitro by light production, and one of these fusions, MPG203, has been further characterized. Transient light production was observed from MPG203 at levels of hydrogen peroxide as low as 10 microM. However, high levels of this toxic oxidizing agent resulted in light suppression, particularly at low bacterial densities. This fusion was also shown to produce light following adhesion to cells of the mouse macrophage cell line J774.2. Furthermore, the response was greatly reduced in the presence of catalase, directly implicating hydrogen peroxide as the eliciting agent and suggesting the involvement of the hydrogen peroxide-induced bacterial stress response in the infection process. Chemiluminescence studies also indicated that inhibition of the respiratory burst may occur as the infection ratio is increased. In addition, the level of light produced from bacteria within individual macrophage cells was shown to vary. PMID- 8423096 TI - An antagonist of platelet-activating factor suppresses endotoxin-induced tumor necrosis factor and mortality in mice pretreated with carrageenan. AB - We found that carrageenan (CAR), that is, sulfated polygalactose, can enhance both lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF) production and the rate of lethality in mice (M. Ogata, S. Yoshida, M. Kamochi, A. Shigematsu, and Y. Mizuguchi, Infect. Immun. 59:679-683, 1991). It has been reported that platelet-activating factor (PAF) antagonists reduce the rate of mortality from endotoxin shock. However, there are few reports regarding the effect of PAF antagonists on TNF production. The aim of the present study is to examine the effect of TCV-309, a new PAF antagonist, on LPS-induced TNF production and mortality in mice pretreated with CAR. ddY mice (6 to 7 weeks old) were injected intraperitoneally with CAR (5 mg per mouse) and were then divided into two groups: mice treated with a PAF antagonist (TCV-309; Takeda Pharmaceutical Co.) and control mice. The mice treated with PAF antagonist received indicated doses of TCV-309 subcutaneously (s.c.) at 30 min before LPS injection, while the control mice received 1 ml of saline s.c. at the same time. All mice were stimulated by intravenous injection of LPS (50 micrograms per mouse) at 24 h after pretreatment with CAR. At intervals after injection of LPS, serum samples were obtained for a TNF assay in which cytotoxicity to L929 cells was measured. TCV-309 both significantly suppressed LPS-induced TNF production and reduced mortality in a dose-dependent manner. When TCV-309 was administered at 30 min before injection of LPS, the effect of TCV-309 on the suppression of TNF activity was at its peak. Treatment with TCV-309 (990 micrograms per mouse) s.c. significantly improved the survival rate after challenge with LPS compared with the survival rate of control mice. Although the 50% lethal dose of LPS was 15 micrograms per mouse for control mice, it increased to 102 micrograms per mouse for mice that were treated s.c. with TCV-309 (990 micrograms per mouse). Even in vitro, TCV-309 also inhibited LPS-induced TNF production in thioglycolate elicited macrophages. It was concluded that PAF plays an important role in endotoxin-induced TNF production and mortality. PMID- 8423095 TI - Neutralization of gamma interferon and tumor necrosis factor alpha blocks in vivo synthesis of nitrogen oxides from L-arginine and protection against Francisella tularensis infection in Mycobacterium bovis BCG-treated mice. AB - Peritoneal cells from Mycobacterium bovis BCG-infected C3H/HeN mice produced nitrite (NO2-, an oxidative end product of nitric oxide [NO] synthesis) and inhibited the growth of Francisella tularensis, a facultative intracellular bacterium. Both NO2- production and inhibition of bacterial growth were suppressed by NG-monomethyl-L-arginine, a substrate inhibitor of nitrogen oxidation of L-arginine, and monoclonal antibodies (MAbs) to gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha). Intraperitoneal injection of mice with BCG increased urinary nitrate (NO3-) excretion coincident with development of activated macrophages capable of secreting nitrogen oxides and inhibiting F. tularensis growth in vitro. Eight days after BCG inoculation, mice survived a normally lethal intraperitoneal challenge with F. tularensis. Treatment of these BCG-infected mice with MAbs to IFN-gamma or TNF-alpha at the time of BCG inoculation reduced urinary NO3- levels to those found in normal uninfected mice for up to 14 days. The same anticytokine antibody treatment abolished BCG-mediated protection against F. tularensis: mice died within 4 to 6 days. Intraperitoneal administration of anti-IFN-gamma or anti-TNF-alpha antibody 8 days after BCG infection also reduced urinary NO3- and abolished protection against F. tularensis. Isotype control (immunoglobulin G) or anti-interleukin 4 MAbs had little effect on these parameters at any time of treatment. IFN-gamma and TNF-alpha were clearly involved in the regulation of macrophage activation by BCG in vivo. Protection against F. tularensis challenge by BCG depended upon the physiological generation of reactive nitrogen oxides induced by these cytokines. PMID- 8423097 TI - Antibacterial activity of lactoferrin and a pepsin-derived lactoferrin peptide fragment. AB - Although the antimicrobial activity of lactoferrin has been well described, its mechanism of action has been poorly characterized. Recent work has indicated that in addition to binding iron, human lactoferrin damages the outer membrane of gram negative bacteria. In this study, we determined whether bovine lactoferrin and a pepsin-derived bovine lactoferrin peptide (lactoferricin) fragment have similar activities. We found that both 20 microM bovine lactoferrin and 20 microM lactoferricin release intrinsically labeled [3H]lipopolysaccharide ([3H]LPS) from three bacterial strains, Escherichia coli CL99 1-2, Salmonella typhimurium SL696, and Salmonella montevideo SL5222. Under most conditions, more LPS is released by the peptide fragment than by whole bovine lactoferrin. In the presence of either lactoferrin or lactoferricin there is increased killing of E. coli CL99 1-2 by lysozyme. Like human lactoferrin, bovine lactoferrin and lactoferricin have the ability to bind to free intrinsically labeled [3H]LPS molecules. In addition to these effects, whereas bovine lactoferrin was at most bacteriostatic, lactoferricin demonstrated consistent bactericidal activity against gram-negative bacteria. This bactericidal effect is modulated by the cations Ca2+, Mg2+, and Fe3+ but is independent of the osmolarity of the medium. Transmission electron microscopy of bacterial cells exposed to lactoferricin show the immediate development of electron-dense "membrane blisters." These experiments offer evidence that bovine lactoferrin and lactoferricin damage the outer membrane of gram-negative bacteria. Moreover, the peptide fragment lactoferricin has direct bactericidal activity. As lactoferrin is exposed to proteolytic factors in vivo which could cleave the lactoferricin fragment, the effects of this peptide are of both mechanistic and physiologic relevance. PMID- 8423098 TI - Secondary Vibrio cholerae-specific cellular antibody responses following wild type homologous challenge in people vaccinated with CVD 103-HgR live oral cholera vaccine: changes with time and lack of correlation with protection. AB - Peripheral blood immunoglobulin A antibody-secreting-cell (ASC) responses are thought to reflect the mucosal immune response to locally presented antigens. We evaluated the ASC response to cholera toxin (CT) and Inaba lipopolysaccharide (LPS) in 26 North American volunteers following immunization with a single oral dose of live attenuated Vibrio cholerae O1 vaccine strain CVD 103-HgR and again upon homologous wild-type challenge with V. cholerae classical Inaba 569B. Challenge occurred at either 7, 30, or 180 days after vaccination. The CT and LPS ASC responses of volunteers following vaccination (83 and 55%, respectively) were similar in magnitude and frequency to those of unvaccinated controls following wild-type challenge (80 and 60%, respectively [0.1 < or = P < or = 0.9]). The responses were primarily immunoglobulin A. Vaccinated volunteers challenged within 30 days of vaccination had reduced or nondetectable CT and LPS ASC responses. Challenge at 6 months resulted in a heightened ASC response to LPS, confirming the existence of mucosal memory. ASC responses to CT upon challenge at 6 months were detectable but not different from that seen following primary immunization, suggesting that secondary ASC responses to different antigens from a single vaccine operate independently. In spite of these variable ASC responses, the vaccine efficacy was 100% following challenge for all vaccinees. V. cholerae specific ASC responses following antigenic reexposure gave information on the presence of mucosal B memory cells but did not correlate with protective immunity. As such, these ASC assays will have limited usefulness for evaluating vaccine responders in vaccine field trials in cholera-endemic areas where prior V. cholerae O1 exposure is unknown. PMID- 8423099 TI - Cellular and humoral immune responses induced in cattle by vaccination with Babesia divergens culture-derived exoantigens correlate with protection. AB - Previous results with the Babesia divergens gerbil vaccination model were extended in studies with cattle. Two calves were vaccinated with culture-derived B. divergens exoantigens, and two others were treated with control supernatant; both preparations were adjuvanted with Quil-A saponin. A parasite-specific humoral response was observed after the first vaccine injection and was boosted by two succeeding vaccine injections. Sera from the two vaccinated calves immunoprecipitated eight major parasitic proteins (with molecular masses ranging between 17 and 110 kDa) whose patterns were close to those observed in gerbil vaccine assays. The cellular immune response, monitored by lymphoproliferation assays, was slightly delayed in comparison with the humoral response; a significant proliferation occurred only after the second vaccine injection. Mononuclear cell proliferation was dose dependent in the presence of (i) lysates of B. divergens-parasitized erythrocytes, (ii) exoantigens of the whole supernatant, or (iii) protective exoantigens of two low-molecular-mass fractions obtained after supernatant gel filtration chromatography. An infectious challenge was administered 3 weeks after the third vaccine injection, with 3.6 x 10(10) B. divergens-parasitized erythrocytes. Erythrocyte count, rectal temperature, and parasitemia of the animals were monitored daily until they returned to initial values. All parameters indicated that the exoantigens induced protection from B. divergens infection for the two vaccinated calves. PMID- 8423100 TI - Identification of a Mycobacterium bovis BCG 45/47-kilodalton antigen complex, an immunodominant target for antibody response after immunization with living bacteria. AB - Increased protection against a virulent challenge with Mycobacterium tuberculosis is induced mainly by a previous immunization with living avirulent mycobacteria, usually Mycobacterium bovis BCG. Only a transient and marginal protection is obtained after immunization with bacterial extracts or dead bacteria. Both living and heat-killed bacteria share a number of common antigens. In order to identify mycobacterial molecules which are dominant antigens during immunization with living bacteria, a two-step selection method was used. Two groups of guinea pigs were immunized either with living or with heat-killed BCG. Sera were then collected and used to select and counterselect antigens present in BCG culture filtrates. Each major fraction eluted from a series of high-pressure liquid chromatography columns (gel filtration, DEAE, and reverse-phase chromatography) was run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred on polyvinylidene difluoride sheets. The molecules present on twin immunoblots were stained with antibodies raised in guinea pigs immunized either with living or with heat-killed BCG. Cross-reactive antigens stained in twin immunoblots were eliminated. Major antigens interacting with antibodies raised after immunization only with living bacteria were further purified. A complex of 45- and 47-kDa major molecules (45/47-kDa complex) was thus identified and further purified. The complex was found to interact only with antibodies present in sera of guinea pigs immunized with living bacteria and not at all with antibodies raised after immunization with dead bacteria. The 45/47-kDa antigen complex molecules were resolved on two-dimensional electrophoresis in three major and seven minor proteins detected with silver staining. All the molecules interacted with the antibodies present in sera of guinea pigs immunized with living BCG. The three major proteins (two at 47 kDa and one at 45 kDa) were amino terminal sequenced. The sequence A-P-E-P-A-P-P-V-P-P-A-A-A-A-P-P-A, which was not previously reported, was the same for these three molecules. By using a competitive enzyme-linked immunosorbent assay, the concentrations of the 45/47 kDa antigen complex were measured in BCG culture filtrates, freeze-dried BCG, and dried heat-killed BCG; they were, respectively, 2, 0.01, and 0.001% of the total mass. The low or very low values compared with the high antibody concentration emphasized the ability of the 45/47-kDa complex delivered through live BCG to trigger an antibody response. PMID- 8423101 TI - Localization of the meningococcal receptors for human transferrin. AB - The interaction between gold-labelled human transferrin (Au-HTF) with live meningococci after growth in vivo or in different in vitro conditions was examined by electron microscopy to localize and quantify the numbers of HTF binding sites on the cell surface. It was clearly demonstrated that HTF binds to the surface of live meningococci (of different serogroups and serotypes) after growth in either iron-sufficient or iron-restricted cultures, although the degree of labelling was always higher (2- to 35-fold) in the latter case. The commensal Neisseria polysaccharea behaved similarly. Ultrathin sections showed that Au-HTF was localized predominantly on the outer membrane of the cells and vesicles, with hardly any internalization. Au-HTF labelling on meningococci was significantly reduced after incubation with unlabelled HTF or with rabbit antiserum containing antibodies against transferrin-binding proteins (TBPs), demonstrating the specificity of the interaction. These sera also blocked binding between HTF and outer membrane proteins on Western immunoblots. Direct evidence of the expression of the TBPs (Western blots) and localization of the HTF receptor (electron microscopy) on in vivo-grown meningococci was obtained from organisms derived without laboratory culturing from the cerebrospinal fluid of a patient. There was considerable cell-to-cell variation in the amount of labelling present on cells of the same sample (in vitro- or in vivo-grown organisms) and between different strains. The degree of binding varied with time of incubation of the cells with Au-HTF. The gold particles frequently formed discrete circles on the cell surfaces of the in vitro-grown organisms; these circles appear to be associated with outer membrane vesicle formation. The results show that the TBPs, which form part of the active components of the HTF receptor(s), are expressed in vivo and are surface exposed and immunogenic and that antibodies against them can interfere with the HTF binding of the meningococcal cells, which may affect iron utilization. This study further supports the concept of regarding the TBPs as future vaccine candidates. PMID- 8423102 TI - Calcium-calmodulin dependence of actin accretion and lethality in cultured HEp-2 cells infected with enteropathogenic Escherichia coli. AB - Infection of cultured HEp-2 cells with enteropathogenic Escherichia coli causes substantial actin accretion at points of bacterial contact and cell death. Loss of viability was delayed by chelating intracellular free calcium. Actin accretion was partially inhibited by preventing elevation of free cytosolic calcium and prevented by treatment with a calmodulin inhibitor. PMID- 8423103 TI - The gamma-hemolysin locus of Staphylococcus aureus comprises three linked genes, two of which are identical to the genes for the F and S components of leukocidin. AB - The Staphylococcus aureus gamma-hemolysin comprises two polypeptides, whereas the gamma-hemolysin locus (hlg) contains three open reading frames. The hlgA and hlgB genes encode the gamma 1 and gamma 2 components, respectively. The HlgB protein (gamma 2) has 27% residue identity with S. aureus alpha-toxin. Surprisingly, hlgB and hlgC are 98.5 and 99.1% identical to the lukF and lukS genes, respectively, encoding the F and S components of the Panton-Valentine leukocidin. PMID- 8423104 TI - Analysis of the immune response in mice following intrauterine infection with the Chlamydia trachomatis mouse pneumonitis biovar. AB - A Swiss Webster white mouse model of salpingitis was used to characterize the immune response following an intrauterine infection with the Chlamydia trachomatis mouse pneumonitis biovar. Western blot (immunoblot) analyses of the serum samples showed that the immunodominant bands corresponded to molecular masses of 72, 60, 42, and 28 kDa and to the lipopolysaccharide. Antibodies to the 60-kDa heat shock protein and to the 60-kDa cysteine-rich protein were detected at 2 and 3 weeks postinfection, respectively. Neutralization was observed in an in vitro assay with serum samples as early as the 3rd day postinfection and remained high for the 7 weeks of observation. The mice were mated in the 7th week following infection. Of the infected experimental mice, 71.4% were found to be either unilaterally or bilaterally infertile, whereas only 27.4% of the noninfected control mice were found to be infertile. PMID- 8423105 TI - Production of mucoid exopolysaccharide during development of Pseudomonas aeruginosa biofilms. AB - Production of mucoid exopolysaccharide by planktonic, chemostat-derived, and adherent Pseudomonas aeruginosa 579 bacteria was separately monitored for 7 days by using a lacZ-algD promoter-reporter gene and assays of total carbohydrate and metabolic activity. Mucoid exopolysaccharide production was transiently elevated following adherence but declined to planktonic levels by day 7. PMID- 8423106 TI - Opsonization of Treponema pallidum is mediated by immunoglobulin G antibodies induced only by pathogenic treponemes. AB - Rabbit antisera to Leptospira interrogans, Borrelia hermsii, and Treponema phagedenis biotype Reiter, reactive to shared spirochetal antigens, failed to enhance phagocytosis of Treponema pallidum by macrophages, while immunoglobulin G to Treponema pallidum subsp. pertenue and Treponema paraluiscuniculi promoted phagocytosis. Opsonic antibodies are directed to pathogen-restricted, not shared spirochetal, antigens. PMID- 8423108 TI - The diagnosis of syncope in the elderly. AB - Given a group of individuals presenting with syncope, the use of the standard diagnostic maneuvers will yield a diagnosis for approximately half. Promising new diagnostic tests offer the prospect of more efficient diagnostic pathways and treatments and demonstrate the need for better clinical trials before they are adopted. PMID- 8423107 TI - Role of antigen-presenting cells in activation of human T cells by the streptococcal M protein superantigen: requirement for secreted and membrane associated costimulatory factors. AB - The requirements for T-cell activation by the streptococcal superantigen (SAg), pepsin-extracted M protein from type 5 streptococci (pep M5), were studied by monitoring Ca2+ influx and cell proliferation. Cells from a pep M5-specific T cell line showed no change in intracellular Ca2+ levels in response to pep M5 when added alone or with freshly isolated autologous antigen-presenting cells (APC). However, after being incubated with pep M5 overnight, the APC secreted soluble factors that together with pep M5 induced a marked increase in intracellular Ca2+ levels in pep M5-specific T cells or freshly isolated, purified T cells. Removal of the SAg from the overnight APC-derived supernatants resulted in loss of the Ca(2+)-mobilizing activity, which was restored within seconds of addition of SAg, suggesting that both the SAg and the soluble factors synergize to induce the Ca2+ influx. Induction of cell proliferation required additional signals inasmuch as the activated APC-derived supernatant failed to synergize with pep M5 to induce the proliferation of purified T cells and required the presence of phorbol myristate acetate for this activity. Metabolically inactive, fixed APC were impaired in their ability to present pep M5 to T cells. Presentation of pep M5 by fixed APC was, however, restored when the APC-derived soluble costimulatory factors were added to the culture. Our data suggest that pep M5-induced activation of T cells is dependent on APC-derived soluble factors and an APC membrane-associated costimulatory molecule(s). These interactions may be important in regulating the in vivo responses to M proteins, could contribute to the severity or progression of infections with Streptococcus pyogenes, and may influence the susceptibility of individuals to its associated nonsuppurative autoimmune sequelae. PMID- 8423109 TI - Making clinical policy explicit. Legislative policy making and lessons for developing practice guidelines. AB - Promulgation of practice guidelines in medicine has increased interest in the structure of clinical policy making. It is argued that with a generic definition of policy as "the rules governing the behavior of individuals or institutions," clinical policy making is analogous to legislative policy making. Practice guidelines emphasize the advantages of making clinical policy more explicit. The structure of legislative policy making has evolved over many years to meet the challenge of making both the policies and the process of policy making explicit. Processes to promulgate clinical policies may be able to exploit this experience to improve clinical policy making and thereby retain control of the process within medicine. Generic steps are outlined for making decisions with incomplete information; synthesis of facts, vested interests, and values; involvement of stakeholders; and implementation of policy. An illustration of the use of the generic steps to make and implement a clinical policy for cesarean birth follows, with evaluations of its impact on the behavior and satisfaction of clinical stakeholders. PMID- 8423110 TI - Noninvasive testing of asymptomatic patients for the detection of silent ischemia after an infarction. A decision analysis. AB - This decision analysis estimates the overall gain in life expectancy and the relative efficacy of predischarge submaximal exercise electrocardiography, ambulatory cardiac monitoring, and thallium-201 scintigraphy for the identification of silent ischemia in asymptomatic postinfarct patients. A small, virtually equal increase in life expectancy can be obtained from any of the noninvasive tests (as compared to no testing). Large differences in life expectancy may result only when the prevalence of residual coronary artery disease and the probability of left-main and three-vessel lesions are high. PMID- 8423111 TI - Ambulatory cardiac monitoring for the evaluation of antiarrhythmic agents. AB - This decision-analysis model assesses ambulatory cardiac monitoring (ACM), ACM followed by exercise testing, and electrophysiologic studies (EPS) in the evaluation and selection of antiarrhythmic agents in postinfarct patients with malignant arrhythmias. With existing data, we find no consistent advantage for one method of drug testing over another, although ACM appears to require fewer resources than does EPS. More patients qualify for EPS, but this fact does not increase the proportion of patients for whom a drug can be identified. These methods may test different aspects of arrhythmia activity and drug response, and sequential use may provide additional benefits. Such benefits must be determined empirically. PMID- 8423112 TI - Accidental injuries of hospitalized patients. A prospective cohort study. AB - Eight hundred and six medical and surgical patients who were hospitalized via the emergency ward were followed over their entire impairment stays and rated in anterograde, double-blind fashion for inpatient incidents (falls, medication errors, other). Injuries were minor but affected 2.2% of admissions, a figure which is strikingly similar to studies in other hospitals. There was a statistical trend toward a higher-than-normal risk of hazardous in-hospital incidents for males age 20 to 40 admitted because of injury and for medically ill females over 60 years old. PMID- 8423113 TI - Standardizing methodologies for economic evaluation in health care. Practice, problems, and potential. AB - There has been an exponential growth in the literature on economic evaluation in health care. As the range and quality of analytical work has improved, economic studies are becoming more influential with health care decision makers. The development of standards for economic evaluation methods would help maintain the scientific quality of studies, facilitate the comparison of economic evaluation results for different health care interventions, and assist in the interpretation of results from setting to setting. However, standardization might unnecessarily stifle methodological developments. This paper reviews the arguments for and against standardization, assesses attempts to date, outlines the main areas of agreement and disagreement on methods for economic evaluation, and makes recommendations for further work. PMID- 8423114 TI - The appropriateness of the use of cardiovascular procedures. British versus U.S. perspectives. AB - To determine whether patients are less likely to receive an inappropriate procedure in countries that devote fewer resources to health care than does the United States, we studied how appropriately coronary angiography and coronary artery bypass surgery were performed in the Trent region of the United Kingdom. The medical records of 320 patients who underwent coronary angiography and 319 who underwent coronary artery bypass surgery in 1987 and 1988 were randomly selected for review. Despite the United Kingdom's more limited use of coronary angiography and coronary artery bypass surgery, a substantial proportion were still performed for less than appropriate reasons, by both U.S. and U.K. criteria. Merely reducing the rate of use of these procedures will not be sufficient to eliminate such inappropriate use. PMID- 8423115 TI - Toward quality assurance in QALY calculations. AB - The utility weights that are used in published scientific articles for assessing benefits in terms of quality-adjusted life-years gained often have a weak theoretical and empirical basis. To a large extent, the weights seem to be used without critical thought and sensible discussion. In a majority of cases, the sensitivity of the results to the weights that have been chosen is not demonstrated, and readers are not provided with information in a way that facilitates independent calculations. This paper calls for a standard for future publications. PMID- 8423116 TI - Defusing technology. Technology diffusion in British Columbia. AB - In order to explore the diffusion of the selected technologies in one Canadian province (British Columbia), two administrative data sets were analyzed. The data included over 40 million payment records for each fiscal year on medical services provided to British Columbia residents (2,968,769 in 1988) and information on physical facilities, services, and personnel from 138 hospitals in the province. Three specific time periods were examined in each data set, starting with 1979-80 and ending with the most current data available at the time. The detailed retrospective analysis of laboratory and imaging technologies provides historical data in three areas of interest: (a) patterns of diffusion and volume of utilization, (b) institutional profile, and (c) provider profile. The framework for the analysis focused, where possible, on the examination of determinants of diffusion that may be amenable to policy influence. PMID- 8423117 TI - Diffusion of new technologies in the treatment of the Medicare population. Implications for patient access and program expenditures. AB - Patterns of diffusion were analyzed for three pairs of technologies: one of each pair represents a very recent innovation, and the other is more established. Tremendous growth in utilization from 1985-89 was documented, primarily due to more patients obtaining access to the technologies. Nevertheless, disturbing differences in levels of use remain between black and white patients. PMID- 8423118 TI - Perspectives on health technology assessment in Latin America. The case of perinatal care in the literature. AB - The possibility of referring to the medical literature to support technology assessments in perinatal care was studied by examining more than 40,000 references contained in the Index Medicus of Latin America for the period 1984 88. A total of 1,074 references addressed perinatal technologies, but with irregular distribution. Information on effectiveness, safety, costs, and social impact of perinatal technologies was extremely limited. These results led to the conclusion that immediate action needs to be taken to create the necessary conditions for health technology assessment in Latin America. PMID- 8423119 TI - Is high-flux dialysis cost-effective? AB - High-flux dialysis is a new method for providing routine-maintenance hemodialysis to patients with end-stage renal disease. It promises to shorten the duration of the dialysis session, but poses potential clinical risks to patients and financial risks to dialysis centers because of the high unit cost of purchasing new dialysis equipment. We retrospectively evaluated the cost-effectiveness of high-flux dialysis compared to conventional dialysis in a hospital-based center. The center provided only conventional dialysis until March 1989, when it initiated high-flux dialysis. The estimated annual costs of treatment were US $31,249 (high-flux) and $32,562 (conventional). The rate of hospital admissions was almost identical in both groups (conventional, 1.29 admissions per year; high flux, 1.24 admissions per year; p = 0.23). Predicted prolongation of life expectancy with high-flux dialysis was significantly higher after statistical adjustment for observable patient characteristics (1.8 to 4.5 years; p < 0.01). The cost-effectiveness ratio was $28,188 per life-year saved for high-flux compared to conventional dialysis. These findings suggest that the added capital expense of purchasing high-flux equipment can be justified from the perspective of its societal cost-effectiveness. PMID- 8423121 TI - Determination of the electrical axis of the heart. AB - The rapid, accurate determination of the electrical axis of a patient's heart is very useful for diagnosis. This determination can be made readily by using six electrocardiographic leads. The procedure for interpreting the data from these leads and determining the electrical axis of the heart is charted. This method uses key leads I and aVF to determine the quadrant where the axis is located and secondary leads II, III, aVR, and aVL to find the axial position within the quadrant. When either I or aVF is isoelectric, only one key lead is used. PMID- 8423120 TI - Assessing the effectiveness of ambulatory cardiac monitoring for specific clinical indications. Introduction. AB - This introduction and the three essays that follow examine ambulatory cardiac monitoring for specific clinical indications. They also examine the ways in which evidence from the literature may be synthesized through the framework of decision analysis to guide its appropriate use and identify areas in which more research is needed. The essays discuss ambulatory cardiac monitoring for evaluation of syncope in the elderly; detection of silent ischemia after a myocardial infarction; and selection of antiarrhythmic drugs for malignant ventricular arrhythmias. PMID- 8423122 TI - Integrate osteopathic principles and practices in postgraduate medical education- now. AB - We issue a call for an "osteopathic medical renaissance" in teaching osteopathic principles and osteopathic manipulative techniques. This article describes a seminar series in the postdoctoral training program at the College of Osteopathic Medicine of the Pacific. We ask osteopathic physicians to enhance their manual dexterity skills to better serve as role models for physicians-in-training. We challenge osteopathic physicians to stimulate a renewed emphasis in teaching distinctive osteopathic medical care. PMID- 8423123 TI - Juvenile myoclonic epilepsy. AB - Juvenile myoclonic epilepsy is a genetically transferred generalized epilepsy that affects approximately 7% of adolescents and adults with epilepsy. Juvenile myoclonic epilepsy always has a myoclonic component alone or in combination with generalized tonic-clonic seizure or absence seizure. The myoclonic component is most prominent on waking or in a drowsy state. The electroencephalogram may be normal between seizures. Life-long treatment with valproate is effective in more than 80% of patients. The author describes the clinical features, diagnosis, and treatment of juvenile myoclonic epilepsy in a typical patient. PMID- 8423124 TI - Peduncular hallucinosis as a transient ischemic attack. AB - Peduncular hallucinosis is characterized by vivid hallucinations associated with organic midbrain disease. In the case reported here, the computed tomography brain scan showed basilar arterial atherosclerotic disease with central and cortical atrophy. Carotid imaging revealed bilateral stenosis. The electroencephalogram and magnetic resonance imaging of the midbrain revealed no abnormalities. The patient's symptoms resolved completely in 48 hours. This is believed to be the first report of peduncular hallucinosis as the manifestation of a transient ischemic attack syndrome. PMID- 8423125 TI - Sample sizes key in interpreting OMT research results. PMID- 8423126 TI - Heterosexual Americans are ignoring safe sex practices. PMID- 8423127 TI - Nephrology/hypertension/fluid and electrolyte disorders. AB - The following annotated bibliography has been developed for the purpose of providing to primary care physicians a handy source of review articles and major studies in the field of nephrology, hypertension, and fluid and electrolyte disorders. Almost all of the articles are review articles of clinical topics that would be of interest to the practicing physician. JAOA will be publishing additional annotated bibliographies in various fields of internal medicine over the next several months. The second in this series will appear in a forthcoming issue of THE JOURNAL. PMID- 8423128 TI - Lead toxicity in children demands DOs' attention. PMID- 8423129 TI - Correlative imaging of pelvic soft tissue sarcomas in children. AB - The authors retrospectively reviewed 15 cases of pelvic soft tissue sarcoma treated at the Children's Hospital of Philadelphia during a recent 6-year period. These patients were treated before the availability of magnetic resonance imaging. The authors compared ultrasonography, computed tomography, and excretory urography/voiding cystourethrography for establishing the diagnosis and following the progression/regression of the disease. Results were correlated with clinical, surgical, and pathologic findings. Although computed tomography and ultrasonography were both capable of characterizing the size and texture of the lesion, computed tomography was superior in defining disease arising from the pelvic sidewall. Both techniques effectively identified residual mass at the tumor site, but neither could differentiate active tumor from inactive tumor, posttherapy inflammation, or fibrosis. Excretory urography/voiding cystourethrography provided no information that could not be gleaned from either ultrasonography or computed tomography. PMID- 8423130 TI - Demonstration of Helicobacter pylori in tracheal secretions. AB - A new gram-negative bacterium, Helicobacter pylori, has been found in gastric secretions. In view of an almost 9% incidence of unidentified gram-negative bacteria in aspiration pneumonia, the authors set out to prove that H pylori could be found in tracheal secretions. Eighteen sequential patients admitted to the intensive care unit who had endotracheal or nasogastric intubation for 24 hours or longer were studied. Of 20 sets of specimens from 18 patients. Helicobacter was recovered from 2(10%) of the endotracheal specimens. The authors conclude that H pylori can gain access to the endobronchial tree and therefore may contribute to the origin of pneumonia due to unidentified gram-negative bacteria. PMID- 8423131 TI - Myofascial release of carpal tunnel syndrome. AB - Current treatment for carpal tunnel syndrome may be ineffective or associated with complications or recurrence. In the case reported here, a myofascial release by the physician combined with the patient's self-stretch reduced pain and numbness and improved electromyographic results. The manipulative approach releases the transverse carpal ligament,-and "opens" or dilates the canal. The patient stretches the wrist, digits, and thumb, including myofascial components. An aggressive, conservative approach lessens the need for surgery in mild to moderate cases. Studies with magnetic resonance imaging may be helpful to document canal size before and after treatment. PMID- 8423132 TI - Environmental effects on the light-harvesting complex of cyanobacteria. PMID- 8423133 TI - Retrotransfer in Escherichia coli conjugation: bidirectional exchange or de novo mating? AB - DNA can be transferred among eubacteria and to plants and fungi by related, plasmid-mediated processes collectively referred to as bacterial conjugation. Conjugation occurs between cells in contact with one another and results in the unidirectional delivery of DNA from a bacterial donor to a recipient. Recent experiments that have reexamined the directionality of DNA flow during conjugation have come to different conclusions, some suggesting that genetic material also flows from recipient cells into the donor and that this process, termed retrotransfer, is likewise directed by donor-encoded functions. Given that bacteria are perhaps united with all living creatures by conjugation, the possibility of gene flow into donor bacteria during conjugation raises interesting evolutionary and biocontainment issues. Here we report that plasmid transmission from bacterial recipients to donors is not a donor-mediated event. Movement of genetic material from recipients to donors was inhibited by streptomycin, which does not inhibit the conjugative donor, indicating that retrotransfer requires gene expression in recipients. Furthermore, retrotransfer was reduced in matings mediated by plasmids that encode strong entry exclusion, to a similar degree as matings between two donors. Therefore we suggest that retrotransfer is in fact newly initiated conjugation between transconjugants and donors. PMID- 8423134 TI - Construction and biochemical characterization of recombinant cytoplasmic forms of the IucD protein (lysine:N6-hydroxylase) encoded by the pColV-K30 aerobactin gene cluster. AB - The aerobactin gene cluster in pColV-K30 consists of five genes (iucABCD iutA); four of these (iucABCD) are involved in aerobactin biosynthesis, whereas the fifth one (iutA) encodes the ferriaerobactin outer membrane receptor. iucD encodes lysine:N6-hydroxylase, which catalyzes the first step in aerobactin biosynthesis. Regardless of the method used for cell rupture, we have consistently found that IucD remains membrane bound, and repeated efforts to achieve a purified and active soluble form of the enzyme have been unsuccessful. To circumvent this problem, we have constructed recombinant IucD proteins with modified amino termini by creating three in-frame gene fusions of IucD to the amino-terminal amino acids of the cytoplasmic enzyme beta-galactosidase. Two of these constructs resulted in the addition to the iucD coding region of a hydrophilic leader sequence of 13 and 30 amino acids. The other construct involved the deletion of the first 47 amino acids of the IucD amino terminus and the addition of 19 amino acids of the amino terminus of beta-galactosidase. Cells expressing any of the three recombinant IucD forms were found to produce soluble N6-hydroxylysine. One of these proteins, IucD439, was purified to homogeneity from the soluble fraction of the cell lysates, and it was capable of participating in the biosynthesis of aerobactin, as determined in vitro by a cell free system and in vivo by a cross-feeding bioassay. A medium ionic strength of 0.25 (250 mM NaCl) or higher was required to maintain the protein in a catalytically functional, tetrameric state.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423135 TI - Correlated physical and genetic map of the Bradyrhizobium japonicum 110 genome. AB - We describe a compilation of 79 known genes of Bradyrhizobium japonicum 110, 63 of which were placed on a correlated physical and genetic map of the chromosome. Genomic DNA was restricted with enzymes PacI, PmeI, and SwaI, which yielded two, five, and nine fragments, respectively. Linkage of some of the fragments was established by performing Southern blot hybridization experiments. For probes we used isolated, labelled fragments that were produced either by PmeI or by SwaI. Genes were mapped on individual restriction fragments by performing gene-directed mutagenesis. The principle of this method was to introduce recognition sites for all three restriction enzymes mentioned above into or very near the desired gene loci. Pulsed-field gel electrophoresis of restricted mutant DNA then resulted in an altered fragment pattern compared with wild-type DNA. This allowed us to identify overlapping fragments and to determine the exact position of any selected gene locus. The technique was limited only by the accuracy of the fragment size estimates. After linkage of all of the restriction fragments we concluded that the B. japonicum genome consists of a single, circular chromosome that is approximately 8,700 kb long. Genes directly concerned with nodulation and symbiotic nitrogen fixation are clustered in a chromosomal section that is about 380 kb long. PMID- 8423136 TI - Characterization of a broad-range disulfide reductase from Streptomyces clavuligerus and its possible role in beta-lactam antibiotic biosynthesis. AB - Streptomyces clavuligerus is a potent producer of penicillin and cephalosporin antibiotics. A key step in the biosynthesis of these beta-lactam compounds is the cyclization of the thiol tripeptide delta-(L-alpha-aminoadipyl)-L-cysteinyl-D valine (ACV) to isopenicillin N by the enzyme isopenicillin N synthase (IPNS). However, bis-ACV, the oxidized disulfide form of the tripeptide, is not a substrate for IPNS. We show here that S. clavuligerus possesses an NADPH dependent disulfide reductase of broad substrate specificity that efficiently catalyzes the reduction of disulfide bonds in bis-ACV and in other low-molecular weight disulfide containing compounds and proteins. The disulfide reductase comprises two protein components, a 70-kDa reductase consisting of two identical subunits, and a 12-kDa heat-stable protein reductant. The structural and functional properties of the disulfide reductase resemble those of the thioredoxin class of oxidoreductases. When the disulfide reductase system is coupled with IPNS, it quantitatively converts bis-ACV to isopenicillin N. These findings suggest that the disulfide reductase may play a role in the biosynthesis of penicillins and cephalosporins in streptomycetes. We also show here that S. clavuligerus lacks glutathione reductase and have previously reported that Streptomyces species do not contain glutathione. This disulfide reductase may therefore be important in determining the thiol-disulfide redox balance in streptomycetes. PMID- 8423137 TI - The product of the hypB gene, which is required for nickel incorporation into hydrogenases, is a novel guanine nucleotide-binding protein. AB - The products of the hyp operon genes are essential for the formation of catalytically active hydrogenases in Escherichia coli. At least one of these auxiliary proteins, HYPB, appears to be involved in nickel liganding to the hydrogenase apoprotein, since mutations in hypB can be phenotypically suppressed by high nickel concentrations in the medium (R. Waugh and D. H. Boxer, Biochimie 68:157-166, 1986). To approach the identification of the specific function of HYPB, we overexpressed the hypB gene and purified and characterized the gene product. HYPB is a homodimer of 31.6-kDa subunits, and it binds guanine nucleotides, with a Kd for GDP of 1.2 microM. The protein displays a low level of GTPase activity, with a kcat of 0.17 min-1. The apparent Km for GTP, as measured in the GTP hydrolysis reaction, was determined to be 4 microM. A chromatography system was established to measure nickel insertion into hydrogenase 3 from E. coli and to determine the effects of lesions in hypB. Nickel appears to be associated only with the processed large subunit of hydrogenase 3 in the wild type, and hypB mutants accumulate the precursor form of this subunit, which is devoid of nickel. The results are discussed in terms of a model in which HYPB is involved in nickel donation to the hydrogenase apoprotein and in which GTP hydrolysis is thought to reverse the interaction between either HYPB or another nickel-binding protein and the hydrogenase apoprotein after the nickel has been released. PMID- 8423138 TI - Autonomous replication of foreign DNA in Histoplasma capsulatum: role of native telomeric sequences. AB - Genetic transformation of the dimorphic pathogenic fungus Histoplasma capsulatum can result in chromosomal integration of the transforming DNA or the generation of multicopy linear plasmids carrying the transforming DNA. We showed previously that Escherichia coli plasmids do not replicate autonomously in H. capsulatum without significant modifications, one of which is the in vivo addition of Histoplasma telomeres at the termini of linear DNA. To address the requirements for autonomous replication in H. capsulatum, we constructed a circular E. coli plasmid containing adjacent inverted stretches of Histoplasma telomeric repeats separated by a unique restriction site. The linearized plasmid bearing telomeric termini was maintained in H. capsulatum without modification other than the addition of more telomeric sequence. We recovered the original plasmid in E. coli after removal of the telomeric termini by using engineered restriction sites. Thus, no special Histoplasma modification or sequence other than the telomeres was needed for autonomous replication in H. capsulatum. Additionally, this plasmid provides a shuttle vector that replicates autonomously in E. coli (as a circular plasmid) and in H. capsulatum (as a linear plasmid). PMID- 8423139 TI - Characterization of dinY, a new Escherichia coli DNA repair gene whose products are damage inducible even in a lexA(Def) background. AB - Bacteriophage Mu dX(Ap lac) was used to isolate a mutation in an Escherichia coli lexA(Def) strain representing a previously undescribed gene (dinY) which does not seem to be under the direct control of LexA. The insertion created a dinY::lacZ fusion in which beta-galactosidase expression required a DNA-damaging treatment (UV irradiation or mitomycin) and activable RecA protein. This strain showed a decreased Weigle reactivation of bacteriophage lambda. However, it was fully inducible for UV mutagenesis. Two-dimensional gel electrophoresis analysis identified two spots absent in the mutant which were both UV inducible only in the presence of activated RecA protein (RecA*). This finding suggests that the dinY::lacZ fusion lies in a gene either that is under the direct control of activated RecA or whose product undergoes RecA*-dependent posttranscriptional/posttranslational modification(s). The dinY gene may also control the expression of some other gene(s) and/or lie in an operon. The fusion was mapped at a position between 41 and 41.5 min on the E. coli chromosome, in the vicinity of the ruv operon. PMID- 8423140 TI - Identification of a putative Bacillus subtilis rho gene. AB - Transposon Tn917 mutagenesis of Bacillus subtilis BD99 followed by selection for protonophore resistance led to the isolation of strain MS119, which contained a single Tn917 insertion in an open reading frame whose deduced amino acid sequence was 56.6% identical to that of the Escherichia coli rho gene product. The insertional site was near the beginning of the open reading frame, which was located in a region of the B. subtilis chromosome near the spoOF gene; new sequence data for several open reading frames surrounding the putative rho gene are presented. The predicted B. subtilis Rho protein would have 427 amino acids and a molecular weight of 48,628. The growth of the mutant strain was less than that of the wild type on defined medium at 30 degrees C. On yeast extract supplemented medium, the growth of MS119 was comparable to that of the wild type on defined medium at 30 degrees C. On yeast extract-supplemented medium, the growth of MS119 was comparable to that of the wild type at 30 degrees C but was much slower at lower temperatures; sporulation occurred and competence was developed in cells of the mutant grown at 30 degrees C. To determine whether the protonophore resistance and sensitivity to low growth temperature resulted from the insertion, a chloramphenicol resistance cassette was inserted into the wild type B. subtilis rho gene of strain BD170; the resulting derivative displayed the same phenotype as MS119. PMID- 8423141 TI - Localizing the replication origin region on the physical map of the Mycoplasma capricolum genome. AB - Four lines of evidence argue that the replication origin of the Mycoplasma capricolum genome lies within the 46-kb BamHI fragment bordered by two BamHI sites of the total of nine BamHI sites that have been located on the physical map (M. Miyata, L. Wang, and T. Fukumura, FEMS Microbiol. Lett. 79:329-334, 1991). First, this fragment lost its labeling in preference to other fragments when log phase cultures were incubated in the presence of chloramphenicol for various times to inhibit the initiation of new rounds of replication and then further incubated with radioactive dTMP to allow DNA elongation to continue. Second, the relative frequencies of various restriction fragments of the genome DNA from exponentially growing cells decreased with increasing distance from the putative origin. Third, preferential labeling occurred when radioactive dTMP was added to cultures of a DNA elongation-defective, temperature-sensitive mutant with a simultaneous temperature downshift. Fourth, the M. capricolum homolog of the dnaA gene, which is located near the replication origin in many other bacteria, was found in the 46-kb fragment. PMID- 8423143 TI - Initial steps in the anaerobic degradation of 3,4,5-trihydroxybenzoate by Eubacterium oxidoreducens: characterization of mutants and role of 1,2,3,5 tetrahydroxybenzene. AB - Chemical mutagenesis and antibiotic enrichment techniques were used to isolate five mutant strains of the obligate anaerobe Eubacterium oxidoreducens that were unable to grow on 3,4,5-trihydroxybenzoate (gallate). Two strains could not transform gallate and showed no detectable gallate decarboxylase activity. Two other strains transformed gallate to pyrogallol and dihydrophloroglucinol but lacked the hydrolase activity responsible for ring cleavage. A fifth strain accumulated pyrogallol, although it contained adequate levels of the enzymes proposed for the complete transformation of gallate to the ring cleavage product. The conversion of pyrogallol to phloroglucinol by cell extract of the wild-type strain was dependent on the addition of 1,2,3,5-tetrahydroxybenzene or dimethyl sulfoxide. This activity was induced by growth on gallate, while the other enzymes involved in the initial reactions of gallate catabolism were constitutively expressed during growth on crotonate. The results confirm the initial steps in the pathway previously proposed for the metabolism of gallate by E. oxidoreducens, except for the conversion of pyrogallol to phloroglucinol. PMID- 8423142 TI - Transcription of the Escherichia coli rrnB P1 promoter by the heat shock RNA polymerase (E sigma 32) in vitro. AB - The P1 promoters of the seven Escherichia coli rRNA operons contain recognition sequences for the RNA polymerase (RNAP) holoenzyme containing sigma 70 (E sigma 70), which has been shown to interact with and initiate transcription from rrn P1 promoters in vivo and in vitro. The rrn P1 promoters also contain putative recognition elements for E sigma 32, the RNAP holoenzyme responsible for the transcription of heat shock genes. Using in vitro transcription assays with purified RNAP holoenzyme, we show that E sigma 32 is able to transcribe from the rrnB P1 promoter. Antibodies specific to sigma 70 eliminate transcription of rrnB P1 by E sigma 70 but have no effect on E sigma 32-directed transcription. Physical characterization of the E sigma 32-rrnB P1 complex shows that there are differences in the interactions made by E sigma 70 and E sigma 32 with the promoter. E sigma 32 responds to both Fis-mediated and factor-independent upstream activation, two systems shown previously to stimulate rrnB P1 transcription by E sigma 70. We find that E sigma 32 is not required for two major control systems known to regulate rRNA transcription initiation at normal temperatures in vivo, stringent control and growth rate-dependent control. On the basis of the well-characterized role of E sigma 32 in transcription from heat shock promoters in vivo, we suggest that E sigma 32-directed transcription of rRNA promoters might play a role in ribosome synthesis at high temperatures. PMID- 8423144 TI - The rightward gas vesicle operon in Halobacterium plasmid pNRC100: identification of the gvpA and gvpC gene products by use of antibody probes and genetic analysis of the region downstream of gvpC. AB - The extreme halophile Halobacterium halobium synthesizes intracellular gas-filled vesicles that confer buoyancy. A cluster of 13 genes on the 200-kb endogenous plasmid pNRC100 has been implicated in the biosynthesis of gas vesicles. Here, we show that two gas vesicle proteins are encoded by genes in the rightward operon, gvpA and gvpC, by Western blotting (immunoblotting) analysis with antibodies directed against LacZ-GvpA and LacZ-GvpC fusion proteins. Our results are consistent with previous data showing that the gvpA gene product is the major gas vesicle protein and demonstrate for the first time that the gvpC gene product is also present in H. halobium gas vesicles. Northern (RNA) blotting analysis showed two RNA species, an abundant 0.35-kb transcript of gvpA and a minor 2.5-kb transcript of gvpAC, and a third gene 3' to gvpAC, named gvpN. The gvpN gene encodes a hypothetical acidic protein with a molecular weight of 39,000 and a nucleotide binding motif. We used a site-directed mutagenesis method involving double recombination in Escherichia coli to insert a kanamycin resistance cassette just beyond the stop codon of gvpN. Introduction of the mutated gene cluster into an H. halobium mutant with a deletion of the entire gas vesicle gene cluster resulted in gas vesicle-positive transformants; this result suggests that gvpN is the last gene of the rightward gas vesicle transcription unit. We discuss the design and utility of the kanamycin resistance cassette for the mutagenesis of other genes in large operons. PMID- 8423145 TI - Xis and Fis proteins prevent site-specific DNA inversion in lysogens of phage HK022. AB - HK022, a temperate coliphage related to lambda, forms lysogens by inserting its DNA into the bacterial chromosome through site-specific recombination. The Escherichia coli Fis and phage Xis proteins promote excision of HK022 DNA from the bacterial chromosome. These two proteins also act during lysogenization to prevent a prophage rearrangement: lysogens formed in the absence of either Fis or Xis frequently carried a prophage that had suffered a site-specific internal DNA inversion. The inversion is a product of recombination between the phage attachment site and a secondary attachment site located within the HK022 left operon. In the absence of both Fis and Xis, the majority of lysogens carried a prophage with an inversion. Inversion occurs during lysogenization at about the same time as prophage insertion but is rare during lytic phage growth. Phages carrying the inverted segment are viable but have a defect in lysogenization, and we therefore suggest that prevention of this rearrangement is an important biological role of Xis and Fis for HK022. Although Fis and Xis are known to promote excision of lambda prophage, they had no detectable effect on lambda recombination at secondary attachment sites. HK022 cIts lysogens that were blocked in excisive recombination because of mutation in fis or xis typically produced high yields of phage after thermal induction, regardless of whether they carried an inverted prophage. The usual requirement for prophage excision was bypassed in these lysogens because they carried two or more prophages inserted in tandem at the bacterial attachment site; in such lysogens, viable phage particles can be formed by in situ packaging of unexcised chromosomes. PMID- 8423146 TI - Presence of two independent chromosomes in the Brucella melitensis 16M genome. AB - Mapping the restriction fragments of the Brucella melitensis 16M genome with a new restriction endonuclease, PacI, which cut the DNA into only eight fragments, indicated that this species contains two unique and independent replicons of about 2,100 and 1,150 kb. Pulsed-field gel electrophoresis of intact DNA revealed two bands migrating the expected distances. These replicons were identified as two unique and independent chromosomes by the presence of rRNA operons and genes for heat shock proteins mapping to separate replicons. PMID- 8423147 TI - Stability and asymmetric replication of the Bacillus subtilis 168 chromosome structure. AB - Chromosomal DNAs from a number of strains derived from Bacillus subtilis 168 were digested with restriction endonucleases NotI or SfiI, and the locations of chromosomal alterations were compared with the recently constructed standard NotI SfiI restriction map (M. Itaya and T. Tanaka, J. Mol. Biol. 220:631-648, 1991). In general, the chromosome structure of B. subtilis 168 was found to be stable, as expected from the genetic stability of this species. DNA alterations, typically deletions, are formed in three limited loci on the chromosome. One of these alterations was characterized as a spontaneous deletion formed between rrn operons, and another occurred as a result of prophage SP beta excision. I found that oriC and terC are not located on precisely opposite sides of the chromosome. Replication in the counter clockwise direction was 196 kb longer than replication in the clockwise direction. The characteristic of length difference is not changed by deletion formation. PMID- 8423148 TI - Polysaccharide synthesis in relation to nodulation behavior of Rhizobium leguminosarum. AB - In this study, we characterized four Tn5 mutants derived from Rhizobium leguminosarum RBL5515 with respect to synthesis and secretion of cellulose fibrils, extracellular polysaccharides (EPS), capsular polysaccharides, and cyclic beta-(1,2)-glucans. One mutant, strain RBL5515 exo-344::Tn5, synthesizes residual amounts of EPS, the repeating unit of which lacks the terminal galactose molecule and the substituents attached to it. On basis of the polysaccharide production pattern of strain RBL5515 exo-344::Tn5, the structural features of the polysaccharides synthesized, and the results of an analysis of the enzyme activities involved, we hypothesize that this strain is affected in a galactose transferase involved in the synthesis of EPS only. All four mutants failed to nodulate plants belonging to the pea cross-inoculation group; on Vicia sativa they induced root hair deformation and rare abortive infection threads. All of the mutants appeared to be pleiotropic, since in addition to defects in the synthesis of EPS, lipopolysaccharide, and/or capsular polysaccharides significant increases in the synthesis and secretion of cyclic beta-(1,2)-glucans were observed. We concluded that it is impossible to correlate a defect in the synthesis of a particular polysaccharide with nodulation characteristics. PMID- 8423149 TI - Conversion of the Salmonella phase 1 flagellin gene fliC to the phase 2 gene fljB on the Escherichia coli K-12 chromosome. AB - The Escherichia coli-Salmonella typhimurium-Salmonella abortus-equi hybrid strain EJ1420 has the two Salmonella flagellin genes fliC (antigenic determinant i) and fljB (determinant e,n,x) at the same loci as in the Salmonella strains and constitutively expresses the fliC gene because of mutations in the genes mediating phase variation. Selection for motility in semisolid medium containing anti-i flagellum serum yielded 11 motile mutants, which had the active fliC(e,n,x) and silent fljB(e,n,x) genes. Genetic analysis and Southern hybridization indicated that they had mutations only in the fliC gene, not in the fljB gene or the control elements for phase variation. Nucleotide sequence analysis of the fliC(e,n,x) genes from four representative mutants showed that the minimum 38% (565 bp) and maximum 68% (1,013 bp) sequences of the fliC(i) gene are replaced with the corresponding sequences of the fljB(e,n,x) gene. One of the conversion endpoints between the two genes lies somewhere in the 204-bp homologous sequence in the 5' constant region, and the other lies in the short homologous sequence of 6, 8, or 38 bp in the 3' constant region. The conversions include the whole central variable region of the fljB gene, resulting in fliC(e,n,x) genes with the same number of nucleotides (1,503 bp) as the fljB gene. We discuss the mechanisms for gene conversion between the two genes and also some intriguing aspects of flagellar antigenic specificities in various Salmonella serovars from the viewpoint of gene conversion. PMID- 8423150 TI - A new type of Alcaligenes eutrophus CH34 zinc resistance generated by mutations affecting regulation of the cnr cobalt-nickel resistance system. AB - Spontaneous mutants that were resistant to zinc were isolated from Alcaligenes eutrophus CH34 containing either the native plasmid pMOL28 or a derivative derepressed for its self-transfer, pMOL50. With the cured plasmid-free derivative of CH34, strain AE104, such mutants were not detected. The mutations, which were shown to be located in the plasmid, increased the level of the nickel and cobalt resistance determined by the cnr locus. The chromate resistance closely linked to the cnr locus was not affected by these mutations. In the Znr mutants, the resistance to zinc and nickel was constitutively expressed. Uptake studies showed that the zinc resistance in a Znr mutant resulted from reduced accumulation of zinc ions in comparison with that in the plasmid-free strain. Reduced accumulation of zinc was also observed to a lesser degree in the parental strain induced with nickel, suggesting that zinc interferes with the Ni2+ and Co2+ efflux system. A 12.2-kb EcoRI-XbaI restriction endonuclease fragment containing the cnr locus was cloned from plasmid pMOL28 harboring the mutation and shortened to an 8.5-kb EcoRI-PstI-PstI fragment conferring resistance to zinc, nickel, and cobalt. The 12.2-kb EcoRI-XbaI fragment was also reduced to a 9.7-kb BamHI fragment still encoding weak resistance to nickel and cobalt but not to zinc. Complementation studies demonstrated the recessivity of the cnr mutations with a Znr phenotype. Such mutations thus allow positive selection of mutants affected in the expression of the cnr operon. PMID- 8423151 TI - Nucleotide sequence of the Serratia marcescens threonine operon and analysis of the threonine operon mutations which alter feedback inhibition of both aspartokinase I and homoserine dehydrogenase I. AB - The nucleotide sequence of the Serratia marcescens threonine operon (thrA1A2BC) was determined. Three long open reading frames were identified; these open reading frames code for aspartokinase I (AKI)-homoserine dehydrogenase I (HDI), homoserine kinase, and threonine synthase, in that order. The predicted amino acid sequences of these enzymes were similar to the amino acid sequences of the corresponding enzymes in Escherichia coli. The AKI-HDI protein is apparently a tetramer composed of monomer polypeptides that are 819 amino acids long. A deletion analysis revealed that the central and C-terminal region was responsible for threonine-resistant HDI activity, a monomeric fragment extending from the N terminus to residue 306 was responsible for threonine-resistant AKI activity, and an N-terminal portion containing 468 residues was responsible for threonine sensitive AKI activity. The thrA(1)1A(2)1 and thrA(1)5A(2)5 mutations of threonine-excreting strains HNr21 and TLr156, which result in the loss of threonine-mediated feedback inhibition of both AKI activity and HDI activity, cause single amino acid substitutions (Gly to Asp at position 330 and Ser to Phe at position 352, respectively) in the central region of the AKI-HDI protein. The thrA1+A(2)2 mutation of strain HNr59, which results in a threonine-sensitive AKI and a threonine-resistant HDI, also causes a single amino acid substitution (Ala to Thr at position 479). PMID- 8423152 TI - Salmonella typhimurium fliG and fliN mutations causing defects in assembly, rotation, and switching of the flagellar motor. AB - FliG, FliM, and FliN are three proteins of Salmonella typhimurium that affect the rotation and switching of direction of the flagellar motor. An analysis of mutant alleles of FliM has been described recently (H. Sockett, S. Yamaguchi, M. Kihara, V. M. Irikura, and R. M. Macnab, J. Bacteriol. 174:793-806, 1992). We have now analyzed a large number of mutations in the fliG and fliN genes that are responsible for four different types of defects: failure to assembly flagella (nonflagellate phenotype), failure to rotate flagella (paralyzed phenotype), and failure to display normal chemotaxis as a result of an abnormally high bias to clockwise (CW) or counterclockwise (CCW) rotation (CW-bias and CCW-bias phenotypes, respectively). The null phenotype for fliG, caused by nonsense or frameshift mutations, was nonflagellate. However, a considerable part of the FliG amino acid sequence was not needed for flagellation, with several substantial in frame deletions preventing motor rotation but not flagellar assembly. Missense mutations in fliG causing paralysis or abnormal switching occurred at a number of positions, almost all within the middle one-third of the gene. CW-bias and CCW bias mutations tended to segregate into separate subclusters. The null phenotype of fliN is uncertain, since frameshift and nonsense mutations gave in some cases the nonflagellate phenotype and in other cases the paralyzed phenotype; in none of these cases was the phenotype a consequence of polar effects on downstream flagellar genes. Few positions in FliN were found to affect switching: only one gave rise to the CW mutant bias and only four gave rise to the CCW mutant bias. The different properties of the FliM, FliG, and FliN proteins with respect to the processes of assembly, rotation, and switching are discussed. PMID- 8423153 TI - Neisseria meningitidis produces iron-regulated proteins related to the RTX family of exoproteins. AB - A monoclonal antibody (A4.85) which reacted with Fe-regulated proteins of Neisseria meningitidis, was used to isolate a lambda gt11 clone from N. meningitidis FAM20. Chromosomal fragments flanking the fragment expressing the A4.85 epitope were cloned, and their DNA sequences revealed a 3,345-bp open reading frame predicting a 122-kDa protein. This gene was named frpA (Fe regulated protein). A computer similarity search of GenBank revealed high levels of similarity to members of the RTX family of cytotoxins, especially in a region of tandem 9-amino-acid repeats. These repeats are found in all members of the RTX family; similar repeats were present 13 times in the predicted FrpA protein. Antigenic relatedness between the meningococcal proteins and the RTX proteins was demonstrated by the reactivity of A4.85 with Escherichia coli hemolysin (HlyA) and Bordetella pertussis adenylate cyclase-hemolysin (CyaA). Similarly, FrpA was recognized by 9D4, a monoclonal antibody directed against B. pertussis CyaA. In addition to the frpA gene, a second gene (frpC) produced a larger RTX-related protein. The frpA and frpC loci were mutagenized in strain FAM20, resulting in the loss of RTX-related proteins. A 120-kDa protein was expressed from the reconstructed frpA gene in E. coli. The biological function of FrpA is unknown, but its similarity to other RTX toxins suggests that it may play an important role in the pathogenesis of meningococcal infection. PMID- 8423154 TI - Insertion derivatives containing segments of up to 16 amino acids identify surface- and periplasm-exposed regions of the FhuA outer membrane receptor of Escherichia coli K-12. AB - The FhuA receptor in the outer membrane of Escherichia coli K-12 is involved in the uptake of ferrichrome, colicin M, and the antibiotic albomycin and in infection by phages T1, T5, and phi 80. Fragments of up to 16 amino acid residues were inserted into FhuA and used to determine FhuA active sites and FhuA topology in the outer membrane. For this purpose antibiotic resistance boxes flanked by symmetric polylinkers were inserted into fhuA and subsequently partially deleted. Additional in-frame insertions were generated by mutagenesis with transposon Tn1725. The 68 FhuA protein derivatives examined contained segments of 4, 8, 12, 16, and 22 additional amino acid residues at 34 different locations from residues 5 to 646 of the mature protein. Most of the FhuA derivatives were found in normal amounts in the outer membrane fraction. Half of these were fully active toward all ligands, demonstrating proper insertion into the outer membrane. Seven of the 12- and 16-amino-acid-insertion derivatives (at residues 378, 402, 405, 415, 417, 456, and 646) were active toward all of the ligands and could be cleaved by subtilisin in whole cells, suggesting a surface location of the extra loops at sites which did not affect FhuA function. Two mutants were sensitive to subtilisin (insertions at residues 511 and 321) but displayed a strongly reduced sensitivity to colicin M and to phages phi 80 and T1. Four of the insertion derivatives (at residues 162, 223, 369, and 531) were cleaved only in spheroplasts and probably form loops at the periplasmic side of the outer membrane. The number and size of the proteolytic fragments indicate cleavage at or close to the sites of insertion, which has been proved for five insertions by amino acid sequencing. Most mutants with functional defects were affected in their sensitivity to all ligands, yet frequently to different degrees. Some mutants showed a specifically altered sensitivity to a few ligands; for example, mutant 511-04 was partially resistant only to colicin M, mutant 241-04 was reduced in ferrichrome and albomycin uptake and showed a reduced colicin M sensitivity, and mutant 321-04 was fully resistant to phage T1 and partially resistant to phage phi 80. The altered residues define preferential binding sites for these ligands. Insertions of 4 to 16 residues at positions 69, 70, 402, 530, 564, and 572 resulted in strongly reduced amounts of FhuA in the outer membrane fraction, varying in function from fully active to inactive. These results provide the basis for a model of FhuA organization in the outer membrane. PMID- 8423155 TI - Characterization of a sucrase gene from Staphylococcus xylosus. AB - The Staphylococcus xylosus gene scrB, encoding a sucrase, has been isolated from a genomic library of S. xylosus constructed in Escherichia coli. The gene was detected by its ability to confer utilization of the glucose and fructose residues of raffinose in an E. coli strain that is not able to metabolize galactose. It was found to reside within a 1.8-kb DNA fragment, the nucleotide sequence of which was determined. One large open reading frame, which is preceded by a ribosome binding site, is encoded on the fragment. Its deduced amino acid sequence yields a protein with a molecular mass of 57.377 kDa which shows significant homology with bacterial sucrose-6-phosphate hydrolases and sucrases. Overexpression of scrB in E. coli by the bacteriophage T7 polymerase promoter system resulted in the production of a protein with an apparent molecular mass of 58 kDa. Disruption of the scrB gene in the S. xylosus genome rendered S. xylosus unable to utilize sucrose. Thus, the ScrB sucrase is essential for sucrose metabolism in S. xylosus. PMID- 8423156 TI - Combinatorial mutagenesis of the lamB gene: residues 41 through 43, which are conserved in Escherichia coli outer membrane proteins, are informationally important in maltoporin structure and function. AB - A new strategy for combinatorial mutagenesis was developed and applied to residues 40 through 60 of LamB protein (maltoporin), with the aim of identifying amino acids important for LamB structure and function. The strategy involved a template containing a stop codon in the target sequence and a pool of random degenerate oligonucleotides covering the region. In vitro mutagenesis followed by selection for function (Dex+, ability to utilize dextrins) corrected the nonsense mutation and simultaneously forced incorporation of a random mutation(s) within the region. The relative importance of each residue within the target was indicated by the frequency and nature of neutral and deleterious mutations recovered at each position. Residues 41 through 43 in LamB accepted few neutral substitutions, whereas residues 55 through 57 were highly flexible in this regard. Consistent with this finding was that the majority of defective mutants were altered at residues 41 to 43. Characterization of these mutants indicated that the nature of residues 41 to 43 influenced the amount of stable protein in the outer membrane. These results, as well as the conserved nature of this stretch of residues among outer membrane proteins, suggest that residues 41 to 43 of LamB play an important role in the process of outer membrane localization. PMID- 8423157 TI - Expression and regulation of Bradyrhizobium japonicum and Xanthobacter flavus CO2 fixation genes in a photosynthetic bacterial host. AB - Calvin cycle carbon dioxide fixation genes encoded on DNA fragments from two nonphotosynthetic, chemolithoautotrophic bacteria, Bradyrhizobium japonicum and Xanthobacter flavus, were found to complement and support photosynthetic growth of a ribulose 1,5-bisphosphate carboxylase-oxygenase (RubisCO) deletion mutant of the purple nonsulfur bacterium Rhodobacter sphaeroides. The regulation of RubisCO expression was analyzed in the complemented R. sphaeroides RubisCO deletion mutant. Distinct differences in the regulation of RubisCO synthesis were revealed when the complemented R. sphaeroides strains were cultured under photolithoautotrophic and photoheterotrophic growth conditions, e.g., a reversal in the normal pattern of RubisCO gene expression. These studies suggest that sequences and molecular signals which regulate the expression of diverse RubisCO genes may be probed by using the R. sphaeroides complementation system. PMID- 8423158 TI - Anaerobic regulation of the adhE gene, encoding the fermentative alcohol dehydrogenase of Escherichia coli. AB - The regulation of the adhE gene, which encodes the trifunctional fermentative acetaldehyde-alcohol dehydrogenase of Escherichia coli, was investigated by the construction of gene fusions and by two-dimensional protein gel electrophoresis. Both operon and protein fusions of adhE to lacZ were induced 10- to 20-fold by anaerobic conditions, and both fusions were repressed by nitrate, demonstrating that regulation is at the level of transcription. Nitrate repression of phi (adhE lacZ) expression, as well as of alcohol dehydrogenase enzyme activity, was partly relieved by a mutation in narL. Mutations in rpoN or fnr had no effect on the expression of adhE. Two-dimensional protein gels demonstrated that increases in the amount of adhE protein correlated with increases in enzyme activity, demonstrating that induction was due to synthesis of new protein, not to activation of preexisting protein. When oxidized sugar derivatives such as gluconate or glucuronate were used as carbon sources, the anaerobic expression of phi (adhE-lacZ) was greatly reduced, whereas when sugar alcohols such as sorbitol were used, the expression was increased compared with expression when glucose was the carbon source. This observation suggested that induction of phi (adhE-lacZ) might depend on the level of reduced NADH, which should be highest with sorbitol grown cells and lowest with glucuronate-grown cells. When phi (adhE-lacZ) was present in a strain deleted for the adhE structural gene, anaerobic expression of phi (adhE-lacZ) was approximately 10-fold higher than in an adhE+ strain. Since the presence of alcohol dehydrogenase would serve to decrease NADH levels, this finding again implies that the adhE gene is regulated by the concentration of reduced NAD. Introduction of a pgi (phosphoglucose isomerase) mutation reduced the anaerobic induction of phi(adhE-lacZ) when the cells were grown on glucose, but had little effect on fructose-grown cells. Pyruvate did not overcome the pgi effect, but glycerol 3-phosphate did, which is again consistent with the possibility that adhE expression responds to the level of reduced NAD rather than to a glycolytic intermediate. PMID- 8423159 TI - Use of synthetic peptides and site-specific antibodies to localize a diphtheria toxin sequence associated with ADP-ribosyltransferase activity. AB - Diphtheria toxin (DT) and Pseudomonas aeruginosa exotoxin A have the same molecular mechanism of toxicity; both toxins ADP-ribosylate a modified histidine residue in elongation factor 2. To help identify amino acids involved in this reaction, sequences in DT that share homology with P. aeruginosa exotoxin A were synthesized and examined for a role in the ADP-ribosyltransferase reaction. By using this approach, residues 32 to 54 of DT were found to define an epitope associated with antibody-mediated inhibition of DT enzyme activity. This lends further support to the notion that residues in this region of DT are involved in the enzymatic reaction. PMID- 8423160 TI - Positive regulation of the Escherichia coli glycine cleavage enzyme system. AB - A new mutation in Escherichia coli, designated gcvA1, that results in noninducible expression of both gcv and a gcvT-lacZ gene fusion was isolated. A plasmid carrying the wild-type gcvA gene complemented the mutation and restored glycine-inducible gcv and gcvT-lacZ gene expression. These results suggest that gcvA encodes a positive-acting regulatory protein that acts in trans to increase expression of gcv. PMID- 8423161 TI - Purification and characterization of SP21, a development-specific protein of the myxobacterium Stigmatella aurantiaca. AB - Stigmatella aurantiaca is a gram-negative bacterium with a complex life cycle, including cellular aggregation resulting in the formation of a characteristic three-dimensional structure, the so-called fruiting body. During fruiting and upon chemical induction of sporulation, a major development-specific protein, SP21, is synthesized. SP21 was purified to homogeneity from the membranous fraction of chemically induced spores. Expression of SP21 was studied with an antiserum raised against the purified protein. PMID- 8423162 TI - Viability of folA-null derivatives of wild-type (thyA+) Escherichia coli K-12. AB - Dihydrofolate reductase (the folA gene product) catalyzes the synthesis of tetrahydrofolate, a key methyl donor in many biosynthetic pathways. Loss of folA had been thought to be lethal to wild-type (thyA+) Escherichia coli. Viable folA null derivatives of thyA+ E. coli were obtained, however, by recombining a folA deletion linked to a Kanr selectable marker into a lambda folA+ phage and using this phage to transduce cells to kanamycin resistance. folA-null strains were slow growing, formed small colonies, and were auxotrophic for thymidine, adenine, methionine, glycine, and pantothenate. PMID- 8423163 TI - The inherent DNase of pyocin AP41 causes breakdown of chromosomal DNA. AB - Pyocin AP41 degrades the chromosomal DNA in sensitive strains of Pseudomonas aeruginosa but has little effect on RNA, protein, and lipid syntheses. In vitro experiments showed that the carboxyl-terminal part of the large subunit of pyocin AP41 carries an inherent DNase that is responsible for its killing action. PMID- 8423164 TI - Evidence for cis-trans isomerization of a double bond in the fatty acids of the psychrophilic bacterium Vibrio sp. strain ABE-1. AB - Vibrio sp. strain ABE-1 was grown in a medium that contained as its stable isotope tracer either [2,2-2H2]cis-9-hexadecenoic or [2,2-2H2]trans-9 hexadecenoic acid. Gas chromatographic-mass spectrometric analysis of the cis-9 hexadecenoic and trans-9-hexadecenoic acid fractions from the cells revealed the formation of an intracellularly isomerized 2,2-2H2-fatty acid which differed from the tracer only in the geometrical configuration of the double bond. This observation shows that cis-trans isomerization without a shift in double-bond position between these two geometric hexadecenoic acid isomers can occur in the cells. PMID- 8423165 TI - The influence of the biarticularity of the gastrocnemius muscle on vertical jumping achievement. AB - Hypotheses concerning the influence of changes in the design of the human musculoskeletal system on performance cannot be tested experimentally. Computer modelling and simulation provide a research methodology that does allow manipulation of the system's design. In the present study this methodology was used to test a recently formulated hypothesis concerning the role of the biarticularity of the gastrocnemius muscle (GAS) in vertical jumping [Bobbert and van Ingen Schenau, J. Biomechanics 21, 249-262 (1988)]. This was done by comparing maximal jump heights for a model equipped with biarticular GAS with a model equipped with a monoarticular GAS. It was found that jump height decreased by 10 mm when GAS was changed into a monoarticular muscle. Thus, the hypothesis formulated by Bobbert was substantiated, although quantitatively the effect is small. Our result differs from that of Pandy and Zajac [J. Biomechanics 24, 1-10 (1991)], who performed similar model calculations. It is shown that the results described by these authors can be explained from the moment-arm-joint-angle relation of GAS at the knee in their model. PMID- 8423166 TI - Nonhomogeneous analysis of epicardial strain distributions during acute myocardial ischemia in the dog. AB - To study the nonuniform mechanical function that occurs in normal and ischemic ventricular myocardium, a new method has been developed and validated. An array of 25 lead markers (approximately 4 x 4 cm) was sewn onto the epicardium of the anterior free wall of the left ventricle in an open-chest, anesthetized canine preparation. Biplane cineradiography was used to track marker positions throughout the cardiac cycle before and during episodes of acute ischemia induced by occlusion of the left anterior descending coronary artery. To estimate two dimensional nonhomogeneous deformations in the region at risk and its border zone with normally perfused tissue, surfaces defined by bicubic Hermite isoparametric finite element interpolation were fitted by least squares to the three dimensional marker coordinates in successive cine frames. Global smoothing functions prevented ill-conditioning in areas of low marker density. Continuous distributions of systolic finite strains referred to the end-diastolic state were obtained under normal and ischemic conditions without the conventional assumption of homogeneous strain analysis. Substantial regional variations in epicardial strains were observed in both the normal and ischemic heart. The method was validated in regions of small to moderate strain variations by comparing the continuous distributions of strain components with piecewise-constant measurements made using marker triplets and homogeneous strain theory. The influence of marker density was examined by recomputing strains from surfaces fitted to subsets of the original array. Further validation of moderate to large strain variations was obtained by simulating a nonuniform distribution of stretch across a planar sheet and computing strains both analytically and using the current method. The new method allows for more comprehensive measurements of distributed ventricular function, providing a tool with which to quantify better the nonhomogeneous function associated with regional ischemia. PMID- 8423167 TI - A computer simulation of the non-Newtonian blood flow at the aortic bifurcation. AB - A two-dimensional numerical model was developed to determine the effect of the non-Newtonian behavior of blood on a pulsatile flow at the aortic bifurcation. The blood rheology was described by a weak-form Casson equation. The successive over-relaxation (SOR) method was used to solve both the vorticity and Poisson equations numerically. It was disclosed that the non-Newtonian property of blood did not drastically change the flow patterns, but caused an appreciable increase in the shear stresses and a slightly higher resistance to both flow separations and the phase shifts between flow layers. PMID- 8423168 TI - An in vivo comparison of anterior tibial translation and strain in the anteromedial band of the anterior cruciate ligament. AB - The objective of this in vivo study was to determine if strain in the anteromedial band (AMB) of the anterior cruciate ligament (ACL) may be predicted by an external measurement of anterior tibial-femoral translation. A Hall effect strain transducer was implanted on the AMB of five human subjects with normal intact ACLs. AMB strain was then measured during anterior shear loading of the tibia relative to the femur, with the knee flexed to 30 and 90 degrees, simulating the loads applied in the Lachman and anterior drawer tests, respectively. The Knee Signature System, a commercially available arthrometer, was used to simultaneously measure anterior tibial translation relative to the femur. The resulting AMB strains and translations during anterior shear loading of the tibia with respect to the femur at 30 and 90 degrees were compared using a regression analysis to determine if AMB strain could be predicted from a measure of anterior tibiofemoral translation at either flexion angle. AMB strain at 150 N anterior shear load at 30 degrees flexion (3.0%) was significantly greater than that at 150 N anterior shear load at 90 degrees flexion (0.9%). During anterior shear loading at 30 degrees flexion, AMB strain correlated with anterior tibial translation (r2 = 0.59). However, there was no significant correlation between AMB strain and anterior tibial translation for anterior shear loading at 90 degrees flexion (r2 = 0.002). Therefore, AMB strain was not accurately predicted from an external measurement of tibial displacement at 90 degrees in this experiment. PMID- 8423169 TI - Impedance matching at arterial bifurcations. AB - Reflections of pulse waves will occur in arterial bifurcations unless the impedance is matched continuously through changing geometric and elastic properties. A theoretical model is presented which minimizes pulse wave reflection through bifurcations. The model accounts for the observed linear changes in area within the bifurcation, generalizes the theory to asymmetrical bifurcations, characterizes changes in elastic properties from parent to daughter arteries, and assesses the effect of branch angle on the mechanical properties of daughter vessels. In contradistinction to previous models, reflections cannot be minimized without changes in elastic properties through bifurcations. The theoretical model predicts that in bifurcations with area ratios (beta) less than 1.0 Young's moduli of daughter vessels may be less than that in the parent vessel if the Womersley parameter alpha in the parent vessel is less than 5. Larger area ratios in bifurcations are accompanied by greater increases in Young's moduli of branches. For an idealized symmetric aortic bifurcation (alpha = 10) with branching angles theta = 30 degrees (opening angle 60 degrees) Young's modulus of common iliac arteries relative to that of the distal abdominal aorta has an increase of 1.05, 1.68 and 2.25 for area ratio of 0.8, 1.0 and 1.15, respectively. These predictions are consistent with the observed increases in Young's moduli of peripheral vessels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423170 TI - A method to determine the range of motion of the ankle joint complex, in vivo. AB - The purpose of this investigation was to develop a method to quantify a range of motion of the ankle joint complex (AJC) in vivo. A six-degree-of-freedom apparatus was constructed to measure the unconstrained active range of motion of the AJC. Extensive tests evaluating the accuracy of the fixture and data collection protocol were conducted. Euler angles similar to dorsiflexion plantarflexion, eversion-inversion, and abduction-adduction were used to quantify the range of motion. Significantly greater (p < 0.05) values were found for plantarflexion, inversion, and abduction of a young group of male subjects (n = 7) compared to an elderly group of male subjects (n = 10). In addition, maximum range of motion values did not necessarily occur from the neutral position. In future, this method can be used to help investigate the relationships between AJC motion and injury, changes in range of motion as a function of age, and design of prosthetic ankle joints. PMID- 8423171 TI - Stress relaxation in native and EDTA-treated bone as a function of mineral content. AB - The relaxation Young's modulus, epsilon(t), of bovine femoral bone was measured as a function of mineral content. Five different specimens with different mineral contents were prepared by the EDTA treatment. The relaxation curves for specimens of different mineral contents were superimposable upon one another by shifting along the log t axis as well as the log epsilon(t) axis. A well-specified master curve of stress relaxation was obtained from a set of relaxation curves for samples with different mineral contents. The mineral content dependence of vertical shift factors, along the log epsilon(t) axis, accorded well with the mineral content dependence of Young's modulus itself, the accordance indicating the plausibility of the vertical shifting procedure. The stress relaxation in bone has been reported to be related to the viscoelastic properties of the collagen matrix. It is considered that the reinforcement of the matrix around the mineral, by the mineral as fillers, increases the average modulus of the matrix and lengthens the characteristic time of the relaxation. This consideration explains the existence of a shift factor along the logt axis and its mineral content dependence. As the structures of bone specimens treated by EDTA are expected to be different from that of normal bone, the conclusion drawn from this experiment cannot be immediately applied to normal bone. It can be, however, concluded that the size of the mineral and also the interface area between mineral and collagen matrix play an important role in the viscoelastic properties of normal bone and the bone specimen treated by EDTA. PMID- 8423172 TI - An assessment of the deflecting effect on human movement due to the Coriolis inertial forces in a space vehicle. AB - Under conditions of prolonged space flight, it may be feasible to restore gravity artificially using centrifugal inertial forces in a spinning vehicle. As a result, the motion of the passengers relative to the vehicle is affected by Coriolis forces. The aim of this study is to propose a theoretical method to evaluate the extent of these effects compared to other inertial or motor forces affecting movement. We investigated typical right upper limb movement in a numerical model with a two-solid-links mechanism, including a spherical joint for the shoulder and a hinge joint for the elbow. The inertial and dimensional characteristics of this model derive from measurements and computations obtained on laboratory subjects. The same is true for the movements assigned to the model. These were inferred from actual recordings of arm movement when the subject presses a button placed in front of him with his index finger. From these relative velocities, the resulting forces and moments applied to the elbow and the shoulder were computed for a 1 rad s-1 rotational speed of transport motion, using classical kinetic relations. The result is that the Coriolis moments are of the same order of magnitude as the corresponding inertial moments and one-tenth of the value of a typical elbow flexion moment. Thus, they should cause a significant disturbance in movement. PMID- 8423173 TI - Changes in the cyclic and static relaxations of the rabbit medial collateral ligament complex during maturation. AB - There is at present no information on how the viscoelastic properties of bone ligament-bone complexes change during maturation. Changes in the static and the cyclic relaxation behaviours of rabbit bone-medial-collateral-ligament-bone complexes at 3, 6, 9 and 12 months of age are described. These ages cover maturation and initial ageing of this complex. The specimens were subject to 30 cycles of uniaxial tension between zero and 0.68 mm extension at 10 mm min-1. Immediately after the cyclic test, the specimens were held at a constant 0.68 mm extension for 1200 s. The results showed that in both the cyclic and the static tests, specimens from 3-month-old animals relaxed significantly more than specimens at 6, 9 or 12 months of age, whereas those 6, 9 and 12 months old were not significantly different from each other. When the results of the cyclic tests were compared to those of the static tests, cyclic relaxation was higher than static relaxation for each of the 6-, 9- and 12-month-old specimens. For specimens 3 months of age, the opposite was observed. An explanation is offered whereby the total relaxation is suggested to be comprised of two parts: a material relaxation and a water flux component. Differing contributions by each of the two components at particular ages can explain the observed results. PMID- 8423175 TI - Orthopaedics at a crossroads. PMID- 8423174 TI - Relative phase quantifies interjoint coordination. AB - This note illustrates by example how expression of joint movement on a phase plane can quantitatively describe multijoint coordination during complex actions. Automatic digitisation of high-speed video records was used to obtain lumbar vertebral, hip, knee and ankle joint angular kinematics in the sagittal plane of a subject performing a symmetric two-handed lifting movement. A consistent proximal-to-distal coordination was illustrated via angle-angle and relative phase angle presentations. During bending to pick up a load, the joints began their movement in the order proximal to distal while the reverse order of joint involvement occurred during extension. Phase angle relationships between joints may provide sufficiently sensitive measurements to identify changes in multijoint coordination induced by alterations in task variables such as (in the case of lifting) object mass, lifting height and load moment. Information regarding multijoint coordination is likely to be important in attempting to understand the respective roles and interaction between the bi and monarticular muscles which are involved in everyday complex actions like lifting. PMID- 8423176 TI - Early failure of short-segment pedicle instrumentation for thoracolumbar fractures. A preliminary report. AB - The results after treatment of fifty-two lumbar and thoracolumbar fractures with Cotrel-Dubousset instrumentation were reviewed as part of an ongoing study. Nineteen patients (average duration of follow-up, fifteen months) had been managed with short-segment pedicle-screw instrumentation. This preliminary report outlines the complications and pitfalls identified during the initial healing phase in this subgroup of patients. There were no neurological or vascular injuries due to placement of the pedicle screws, but ten patients had some form of failure of the fixation during the early period of healing. Failure of the fixation was manifested in three ways: progressive kyphosis secondary to the bending of screws (six patients), kyphosis secondary to osseous collapse or vertebral translation without bending of the hardware (three patients), and segmental kyphosis after a caudad screw in the lumbar construct broke (one patient, who had had a combined instrumentation for multiple fractures). Untreated anterior instability, and pre-stressing of the screws when the rods were contoured in situ, resulted in a high rate of failure. The high rate of failure of the hardware associated with this fixation construct suggests that posterior screw fixation alone may not be adequate when Cotrel-Dubousset instrumentation is used for short-segment lumbar arthrodeses. Bent screws or measurable kyphosis did not always herald a clinical failure, but patients who had progressive kyphosis of more than 10 degrees had substantially more pain than did those who had little or no progression. The results reported here are preliminary, and speculation as to the importance of these findings and as to the long-term outcome in these patients would be premature.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423177 TI - Spinal mobility and deformity after Harrington rod stabilization and limited arthrodesis of thoracolumbar fractures. AB - The results were reviewed for thirty patients in whom a thoracolumbar fracture had been treated with a Harrington rod placed three segments cephalad and two or three segments caudad to the injured vertebra and a short arthrodesis with removal of the Harrington rod after at least twelve months. The patients were evaluated with clinical examinations, lateral radiographs made with the spine in flexion and extension, and questionnaires at a median of thirty-two months after the stabilization. All of the arthrodeses were successful. The twenty patients who had a burst or compression fracture and who had posterior instrumentation lost only an average of 9 degrees of sagittal correction before attaining a solid fusion. For all thirty patients, the average motion in the sagittal plane at the vertebrae that had been spanned by the Harrington rod but not included in the arthrodesis ranged from 9 degrees at the third and fourth lumbar level to 2 degrees at the eighth and ninth thoracic level. The average total motion between five vertebrae that had been spanned by the rod but did not have an arthrodesis was 24 degrees; between four vertebrae, 23 degrees; and between three vertebrae, 19 degrees. In this series of thoracolumbar fractures treated with a longer Harrington rod than needed to span the level of the arthrodesis and a short arthrodesis, there was a documented return of motion to segments that had been previously spanned by the instrumentation but not included in the arthrodesis and good or excellent functional results over-all. PMID- 8423178 TI - Regulation of growth-plate chondrocytes by insulin-like growth-factor I and basic fibroblast growth factor. AB - A study was performed in order to investigate the possible functional roles of insulin-like growth-factor I (IGFI) and basic fibroblast growth factor (bFGF) in the regulation of mitotic and metabolic activity of growth-plate chondrocytes. Chondrocytes from the distal radial growth plates of calves and the costal physeal cartilage of rats were exposed to these factors, individually and in combination, in primary monolayer culture, to assess their effects. The data showed that bFGF had both a greater potency and a greater efficacy as a mitogen for bovine growth-plate chondrocytes than did IGF-I. The maximum incorporation of 3H-thymidine by bFGF was 8.3 times that in serum-free (control) cultures; the maximum stimulation of incorporation by IGF-I was 2.5 times that in the control medium. In contrast, IGF-I stimulated a maximum incorporation of 35S-sulfate into glycosaminoglycans that was 2.6 times that in the IGF-I serum-free control cultures, while bFGF had no effect or was mildly inhibitory. When used together, these two factors acted synergistically. Incorporation of 3H-thymidine was more than two times greater than the sum of the effects of the growth factors when used alone and 20.5 times greater than that of the growth factor-free control cultures. Physeal chondrocytes from six-day-old rats were mitotically more responsive to bFGF than to IGF-I, but they were more responsive to IGF-I when they had been derived from twenty-eight-day-old rats. Interaction between bFGF and factors in the serum enhanced the mitotic activity of the rat chondrocytes, but bFGF did not interact with IGF-I under the same experimental conditions. In the presence of bFGF, there was a reduction in the stimulation by IGF-I of incorporation of 35S-sulfate and a decrease in the percentage of chondrocytes containing alkaline phosphatase. These growth factors also influenced cellular morphology in culture. In the presence of IGF-I or serum, the rat chondrocytes manifested the polygonal morphology typical of chondrocytes in culture, while bFGF promoted a more elongated spindle shape. Removal of bFGF and replacement by IGF-I restored the polygonal morphology, indicating that this transition is reversible. PMID- 8423179 TI - Deformity of the calcaneocuboid joint in patients who have talipes equinovarus. AB - A retrospective analysis was done of the records and radiographs of 100 club feet (in sixty-six patients) that had been consecutively treated with an operation. Twenty-six feet (26 per cent) had had a Grade-II deformity of the calcaneocuboid joint, as determined with a radiographic classification that had been developed on the basis of the degree of medial displacement of the cuboid. When the calcaneocuboid joint is in normal alignment, the central point of the cuboid ossification center lies on the mid-longitudinal axis of the calcaneus; when there is a Grade-I deformity, the mid-point of the cuboid ossification center lies lateral to the medial tangent but medial to the longitudinal axis of the calcaneus; and when there is a Grade-II deformity, the central point of the cuboid lies on or medial to the medial tangent of the calcaneus. Although a Grade I deformity of the calcaneocuboid joint need not be corrected, a Grade-II deformity should be treated with release of the calcaneocuboid joint, which in this series was performed in conjunction with a complete subtalar release (including a talonavicular release). Sixteen of the twenty-six feet that had a Grade-II deformity had a complete release of the calcaneocuboid joint at the time of the operation; the release was not done in the remaining ten feet, some of which were operated on early in the series, before the importance of the deformity at the calcaneocuboid joint had been recognized.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423180 TI - Proximal tibial osteotomy. A critical long-term study of eighty-seven cases. AB - Eighty-seven valgus osteotomies of the tibia were performed in seventy-three patients for osteoarthrosis of the medial compartment of the knee; the median follow-up was ten years (range, three to fourteen years). The data were subjected to univariate and multivariate statistical analysis and to survivorship analysis. For these calculations, the end-point of failure was defined as an arthroplasty of the knee, and additional calculations were performed with the end-point defined as the performance of an arthroplasty or moderate or severe pain in patients who had declined an arthroplasty. None of the many risk factors that were evaluated could be found to be associated with the duration of survival, except for relative weight and angular correction. The median loss of correction after the osteotomy was 1 degree. If, at one year after the operation, the valgus angulation was 8 degrees or more, or if the patient's weight was 1.32 times the ideal weight or less, the probability of survival five years thereafter was at least 90 per cent and the probability ten years thereafter was at least 65 per cent. However, when valgus angulation at one year was less than 8 degrees in a patient whose weight was more than 1.32 times the ideal weight, the rate of survival decreased to 38 per cent five years thereafter and to 19 per cent ten years thereafter. There is a considerable risk of failure of a proximal tibial osteotomy if the alignment is not overcorrected to at least 8 degrees of valgus angulation and if the patient is substantially overweight. PMID- 8423181 TI - Motor recovery after arthroscopic partial meniscectomy. Analyses of gait and the ascent and descent of stairs. AB - We studied motor recovery as shown by locomotor activities after arthroscopic partial medial meniscectomies in seventeen men who were twenty-five to forty-nine years old. The patients were evaluated before the operation and two, four, and eight weeks after the operation. Control values were obtained from twenty-two healthy men whose ages, weights, and heights were similar to those of the patients who had had a meniscectomy. Motion of the hip, knee, and ankle in the sagittal plane and the electromyographic activities (as measured with surface electrodes) in five muscles were recorded while each subject walked on a level walkway and then ascended and descended stairs at free speeds. The results showed that meniscal tears affect the motor-control mechanisms involved in the submaximum locomotor activities that were studied and that these abnormalities may persist for as long as eight weeks after a meniscectomy. PMID- 8423182 TI - The natural history of heterotopic ossification in patients who have fibrodysplasia ossificans progressiva. A study of forty-four patients. AB - Forty-four patients who had fibrodysplasia ossificans progressiva responded by mail to a questionnaire regarding the age at the onset of heterotopic ossification at fifteen commonly involved anatomical sites. The average age of the patients when they responded to the questionnaire was twenty-seven years (range, three to sixty-nine years). The average age at the onset of ossification was five years (range, birth to twenty-five years). The most common sites of early heterotopic ossification were the neck, spine, and shoulder girdle. Thirty five (80 per cent) of the patients had had some restrictive heterotopic ossification by the age of seven years. By the age of fifteen years, forty-two (more than 95 per cent) of the patients had severely restricted mobility of the upper limbs. In these patients, heterotopic ossification proceeded in a direction that was axial to appendicular, cranial to caudad, and proximal to distal; this pattern appeared typical for fibrodysplasia ossificans progressiva. PMID- 8423183 TI - Measurement of intracompartmental pressure: a comparison of the slit catheter, side-ported needle, and simple needle. AB - An experimental model of acute compartment syndrome involving the anterolateral compartment of the hindlimb in dogs was used to compare three methods of measurement of intracompartmental pressure: the simple-needle technique, use of the slit catheter, and use of the side-ported needle. No statistical difference was found between the values obtained with the slit catheter and those obtained with the side-ported needle; the mean difference was 1.4 millimeters of mercury throughout the range of compartment pressures that were measured. The side-ported needle appeared to be as accurate as the slit catheter for the measurement of compartment pressures (p = 0.355, 1-beta = 0.9). The values obtained with use of the simple needle were consistently higher than those obtained with the other two methods (p < 0.001): an average of 18.3 millimeters of mercury higher than the values measured with the slit catheter and 19.3 millimeters of mercury higher than those measured with the side-ported needle. Clinically, the side-ported needle or the slit catheter can be used to obtain accurate measurements of compartment pressure. Use of the simple 18-gauge needle is not recommended for this purpose. PMID- 8423184 TI - The natural history and long-term follow-up of Scheuermann kyphosis. AB - Sixty-seven patients who had a diagnosis of Scheuermann kyphosis and a mean angle of kyphosis of 71 degrees were evaluated after an average follow-up of thirty-two years (range, ten to forty-eight years) after the diagnosis. All sixty-seven patients completed a questionnaire; fifty-four had a physical examination and radiographs; fifty-two, pulmonary function testing; and forty-five, strength testing of the trunk muscles. The results were compared with those in a control group of thirty-four subjects who were matched for age and sex. The patients who had Scheuermann kyphosis had more intense back pain, jobs that tended to have lower requirements for activity, less range of motion of extension of the trunk and less-strong extension of the trunk, and different localization of the pain. No significant differences between the patients and the control subjects were demonstrated for level of education, number of days absent from work because of low-back pain, extent that the pain interfered with activities of daily living, presence of numbness in the lower extremities, self-consciousness, self-esteem, social limitations, use of medication for back pain, or level of recreational activities. Also, the patients reported little preoccupation with their physical appearance. Normal or above-normal averages for pulmonary function were found in patients in whom the kyphosis was less than 100 degrees. Patients in whom the kyphosis was more than 100 degrees and the apex of the curve was in the first to eighth thoracic segments had restrictive lung disease. Five patients had an unexplained, mildly abnormal neurological examination. Mild scoliosis was common; spondylolisthesis was not observed. PMID- 8423185 TI - Long-term results of staple capsulorrhaphy for anterior instability of the shoulder. AB - The results of 204 open staple capsulorrhaphies, performed consecutively as treatment for recurrent anterior instability of the shoulder in 192 patients, were reviewed after an average of ten years (range, two to twenty years). The operation had been performed for recurrent dislocations in 88 per cent of the shoulders and for recurrent subluxations in the remaining 12 per cent. Postoperative instability--dislocation or subluxation--occurred in 22 per cent of the shoulders and increased in frequency logarithmically with the duration of follow-up. In more than half of these shoulders, the episodes of postoperative instability were recurrent. In one-third of the shoulders, the stapling had been combined with a Putti-Platt procedure; in the others, a muscle-splitting approach had been used. The rate of recurrent instability was 8 per cent in the shoulders in which a Putti-Platt procedure had been added and 29 per cent in the shoulders that had been treated by stapling alone. The difference was significant (p = 0.002). Loosening or migration of a staple, or penetration of the articular cartilage by a staple, occurred in twenty-four shoulders (12 per cent); the staple was removed from eighteen of them. There was no significant difference in the rate of loosening or migration between non-barbed and barbed staples (p = 0.92). Pain, physical restrictions, and osteoarthrosis were more frequent in patients who had complications associated with a staple. Although most of the patients (84 per cent of the shoulders) thought that they had benefited from the operation, approximately half (51 per cent of the shoulders) had pain and approximately half (50 per cent of the shoulders) said that the shoulder was sufficiently different from normal to affect the quality of life. Problems with the shoulder that had not been present before the operation caused several patients (5 per cent of the shoulders) to change occupations. The average ranges of internal and external rotation were slightly reduced. The subjective and objective results after the stapling procedure were not as good as previous reports have suggested, and we no longer recommend staple capsulorrhaphy for anterior instability of the shoulder, even when it is augmented by a Putti-Platt procedure. PMID- 8423186 TI - A syndrome of dislocated hips and radial heads, carpal coalition, and short stature in Puerto Rican children. AB - An orthopaedic syndrome that apparently had not been reported previously was identified in twenty-three children. Characteristics shared by all twenty-three children included Hispanic descent, residence in Puerto Rico, bilateral dislocation of the hip, dislocated radial heads, short stature, and other osseous anomalies. Twelve dislocated hips in six patients were not treated. All of these hips were functioning satisfactorily at the time of the review, but only four of the children had reached skeletal maturity. Sixteen hips in eight patients remained reduced after closed reduction. Of these eight patients, the four who were skeletally immature at the time of the review had a satisfactory result, and the four who were skeletally mature had an unsatisfactory result because of discomfort or fibrous ankylosis. Eighteen hips in nine patients were treated with a reduction augmented by some form of operation. All of these hips redislocated. Of the forty-six elbows in the twenty-three children, thirty-three were dislocated, as seen clinically and radiographically; eight were normal, both clinically and radiographically; and there was dysplasia at the radiocapitellar articulation of the remaining five. Twenty of the twenty-three children were found to have carpal coalitions. Fourteen children had scoliosis, and five of them were managed with spinal arthrodesis and correction. Three patients had an anomaly of the cervical spine, with one deformity causing symptoms and signs that were treated with decompression. Eight patients had talipes cavus bilaterally, which was not treated. PMID- 8423187 TI - Osteofibrous dysplasia in the newborn. Report of a case. PMID- 8423188 TI - Extensive osteolysis behind an acetabular component that was well fixed with cement. A case report. PMID- 8423189 TI - Fracture of the olecranon through a persistent physis in an adult. A case report. PMID- 8423190 TI - Premature monomelic physeal closure. A case report. PMID- 8423191 TI - Alternatives to reimplantation for salvage of the total knee arthroplasty complicated by infection. PMID- 8423192 TI - The clinical results and basic science of total hip arthroplasty with porous coated prostheses. PMID- 8423193 TI - Treatment of instability of the shoulder with an exercise program. PMID- 8423194 TI - Experimental models of kidney tumors. AB - We here present in outline some outstanding results on the animal models of renal tumors submitted to the highest attention, which include two kinds of epithelial neoplasms: those developed from the epithelium lining each the renal tubules (renal cell tumors) and pelvis; the mesenchymal tumor of rat; and tumors with embryonal appearance: the nephroblastoma as well as the variant of it known as the estrogen-induced tumor in the hamster. The review deals with methods for tumor induction and the pathobiology of tumors, the latter mainly focused on the identification of the precise types of renal cells committed in tumor origin. The existence of closely related counterparts of these animal tumors in man is also emphasized, by representing a situation of great potentiality, already exploited, to improve understanding of both the nature and development of renal tumors in man. PMID- 8423195 TI - Breaks in DNA accompany estrogen-receptor-mediated cytotoxicity from 16 alpha[125I]iodo-17 beta-estradiol. AB - Strategies for diagnosis and therapy in which sex steroid receptor ligands serve as carriers for radionuclides are attractive because a high incidence of carcinomas of the female genital tract and the breast that are seen clinically have an abundant expression of one or more of the receptor proteins. A radiohalogenated estrogen receptor (ER) ligand, 16 alpha-[123I]iodo-17 beta estradiol [123I]E, has met clinical criteria for receptor-mediated diagnostic imaging. Its [125]I-labeled sister nuclide derivative [125I]E decays by orbital electron capture with emission of very-low-energy (Auger) electrons, which gives this latter radiohalogen the potential to serve in pharmaceuticals for radiotherapy; as examples, [125I]deoxyuridine, when incorporated into the DNA molecule, or [125I]E, when bound to the receptor within ER-rich tumor cells, are both cytotoxic in vitro. Whereas the mechanisms and subcellular changes that accompany the cytotoxicity from [125I]deoxyuridine are well documented in the form of aberrations and breaks in the cellular DNA, the effects at the subcellular level causing the cytotoxicity of the sex steroid receptor ligand [125I]E have not been characterized and are the focus of our study. We found that in a standard colony-forming assay the addition of [125I]E to the cultures decreased the survival rate of ER-positive MCF-7 cells in a dose-dependent manner. The decreased survival rate was prevented by the addition of competing excess radioinert ER ligand (diethylstilbestrol); [125I]E did not reduce survival in ER-negative MCF-7 cells. The [125I]E-induced and ER-mediated cytotoxicity was accompanied by aberrations in the DNA components of the nuclei of the cells. These included chromatid and chromosome breaks, gaps, and tri-radial chromosome formation. Our findings add plausibility and credence to the notion that the cytotoxicity imparted by Auger-electron-emitting radioligands for sex steroid receptors is in part attributable to radiodecay that causes double-stranded breakage of DNA. PMID- 8423196 TI - Treatment of the rat R-1 rhabdomyosarcoma with methotrexate and radiation; effects of timing on cell survival and tumour growth delay. AB - The interaction of radiation (10 Gy 300-kV X-rays) and methotrexate (MTX; 3 x 10 mg/kg at 3.5-h intervals) was investigated with respect to effects on cell survival and tumour regrowth of the transplantable rat R-1 rhabdomyosarcoma. The treatment with MTX alone caused acceptable toxicity and no lethality. On day 3 after treatment with MTX alone a maximum decrease in the fraction of clonogenic cells was observed, which is in accordance with data on MTX concentrations in tumour tissue, indicating that MTX is active in the tumour for at least 3 days after injection. The clonogenic capacity after combined treatments, i.e. MTX before or after radiation, was assessed 3 days after the MTX administration. The fractions of clonogenic cells determined after combined therapy with intervals of up to 4 days were not significantly different from those expected on the basis of simple addition of the effects from individual treatments. However, the excess growth delay was positive at specific intervals (6-8 days after X-rays plus MTX and 5-6 days after MTX plus X-rays), whereas negative excess delays were observed when the two treatments were separated by less than 3 days. It is concluded that expectations with respect to clinical application of the combination must be modest in view of the short duration of favourable intervals and the observed absence of synergistic effects with respect to cell killing. The discrepancy between the two assays indicates that erroneous conclusions can be obtained if one endpoint only is assessed. PMID- 8423197 TI - Production and characterization of monoclonal antibodies to N7-phenylguanine. AB - N7-Phenylguanine, a base adduct possibly formed after arylation of DNA by benzene oxide, the first reaction metabolite during benzene metabolism, was synthesized in our laboratory and used as reference for the production and characterization of monoclonal antibodies. 2-Hydroxymethyl-7-phenylhypoxanthine, a molecule structurally similar to N7-phenylguanine, was coupled by a linker molecule to different carrier proteins. The resulting conjugate was used to immunize BALB/c mice, the spleen cells of which were fused with mouse P3X63-Ag8.653 myeloma cells to obtain monoclonal antibodies. Several hybridoma lines were cultivated in defined media and characterized as to sensitivity and specificity by an enzyme linked immunosorbent assay (ELISA). Competitive ELISA demonstrated that all antibodies showed a very high affinity for N7-phenylguanine but had a lower affinity towards various other samples including N7-chlorophenylguanines and C8-, N2-and O6-phenylguanine. As little as about 20 pg N7-phenylguanine could be detected with one of the most sensitive antibodies, CE6/G11, with a colorimetric end point while the detection limit could be lowered to about 10 pg N7 phenylguanine when a fluorescent end point was used. The detection limit of other methods used to determine N7-phenylguanine so far is 10 ng for gas chromatography/mass-spectrometry and 1 ng for high-pressure liquid chromatography. Thus the use of specific monoclonal antibodies seems to be the most sensitive method for the detection of N7-phenylguanine. PMID- 8423198 TI - Transient cisplatin-resistant murine fibrosarcoma cell characterized by increased metallothionein content. AB - Cisplatin-resistant mouse fibrosarcoma cells, SSK-R, were isolated after short and low-dose drug treatment of the sensitive SSK cells in vitro. These SSK-R sublines exhibit up to sevenfold cisplatin resistance and are characterized mainly by an increased metallothionein content. Loss of drug resistance after about 140-180 cell divisions in drug-free medium coincides with loss of metallothionein content. The glutathione level is the same in the sensitive and resistant sublines; inhibition of glutathione synthesis by buthionine sulphoximine enhances the sensitivity in both cells lines by a factor of 2.7. The resistant sublines are not cross-resistant to radiation; a radiation exposure followed immediately by cisplatin treatment results in an additive effect. The cellular cisplatin content is slightly reduced in SSK-R2 cells and it remains at this level also upon loss of drug sensitivity. PMID- 8423199 TI - Phase II trial of zeniplatin (CL 286,558), a new patinum compound, in patients with advanced ovarian cancer previously treated with organoplatinum-based therapy. AB - There is a critical need to find new chemotherapeutic agents that are active in platinum-refractory ovarian cancer. A phase II trial of zeniplatin (CL 286,558), a third-generation platinum compound, was conducted in 31 patients with advanced ovarian cancer to examine the safety and activity of the agent when used as a salvage treatment in individuals previously exposed to organoplatinum-based therapy. In general the drug was well tolerated, with moderate emesis and bone marrow suppression being observed in most patients. An unexpected side-effect was significant fever, of unknown etiology, which was noted in 16% of patients. Out of 20 patients, 2 (10%; 95% confidence intervals: 1%-32%) with clinically defined platinum-refractory disease achieved a partial response. Unfortunately, although we have defined definite but modest activity for zeniplatin in platinum refractory ovarian cancer, further development of this drug has been discontinued because of the severe renal toxicity observed in other clinical trials of this cytotoxic agent. PMID- 8423200 TI - Increasing rates of DNA single-strand breaks in lymphocytes of clinical personnel handling cytostatic drugs. AB - A total of 27 persons, working in cancer stations with exposure to cytostatics, and 40 healthy control persons were examined for DNA single-strand breaks in peripheral lymphocytes. Non-smoking personnel from cancer stations were found to have an increased rate of DNA single-strand breaks compared to the non-smoking control subjects. In the case of smokers an increased rate of DNA single-strand breaks could be recorded for those working in cancer stations as well as with the controls. DNA single-strand breaks indicate reversible damage to DNA. As DNA repair is not perfect in every case, an increased number of DNA single-strand breaks leads to irreversible DNA damage. PMID- 8423202 TI - International Neuropsychological Society 21st Annual Meeting. Galveston, Texas, February 24-27, 1993. Abstracts. PMID- 8423201 TI - Symposium on human tumour viruses. PMID- 8423203 TI - Segregation of immunoglobulin heavy chain constant region genes in multiple sclerosis sibling pairs. PMID- 8423204 TI - HTLV-1 antibody class and subclass distribution in African TSP and control populations. AB - The humoral immune responses in 44 sera from HTLV-1 seropositive African subjects were compared. The sample population was composed of 12 patients with HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP), 12 patients with other neurological conditions and 20 asymptomatic carriers. Samples HTLV-1 antigens were tested against all immunoglobulin classes and IgG subclasses, using the Western blot technique with polyclonal and monoclonal antibodies. Whilst IgG reacted with gag, env and tax products for the three groups studied, IgM and IgA were found to react more frequently with HTLV-1 in HAM/TSP patients. For these patients, IgM and IgA were particularly directed against tax and env proteins. Among IgG subclasses, IgG1 was most sensitive to gag, env and tax products reacting in similar proportions in all three groups. IgG2 and IgG4 were apparently not involved. IgG3 was most responsive in HAM/TSP patients. These data are similar to those observed in AIDS patients, LAS and HIV asymptomatic carriers and emphasize the role of HTLV-1 in HAM/TSP. PMID- 8423205 TI - Vimentin-crossreactive antibodies induce cell death in primary cultures of embryonic spinal cord. AB - Sera from guinea pigs inoculated with spinal cord gray matter were cytotoxic for embryonic day 14 (ED14) primary spinal cord cell cultures. All cell types including astroglia and neurons were affected. Immunoglobulin (IgG) from inoculated animals and biologically active complement proteins were both identified as necessary components for the observed cytotoxicity. Adsorption with cytoskeletal constituents removed the cytotoxic effect. Adsorption with purified vimentin substantially reduced cytotoxicity, while adsorption with glial fibrillary acid protein (GFAP) had no cytoprotective effects. Antiserum from vimentin-immunized guinea pigs was also toxic to spinal cord cell cultures. Antibody and complement-mediated toxicity appeared to result from interaction with vimentin bound to the surface of embryonic cultured glia and neurons, although interaction with a cross-reacting epitope could not be definitely excluded. Cytotoxic vimentin-directed antibody was not noted in control sera, and the presence of this antibody in guinea pig and human sera did not correlate with the clinicopathological state. PMID- 8423206 TI - Immunological changes in the MPTP-induced Parkinson's disease mouse model. AB - The role of the central dopaminergic system in modulating immune response is not completely established. We examined the influence of central dopamine depletion on selected parameters of immune functions in 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) treated and untreated mice. IgM antibody production of splenocytes to sheep red blood cells was reduced in MPTP-treated mice (P < 0.001). Proliferation of splenocytes in response to a wide range of mitogen concentrations (Concanavalin A, phytohaemagglutinin, lipopolysaccharide) was also significantly diminished in MPTP-treated mice. Production of migration inhibition factor (MIF) was diminished only in low mitogen concentration. Our results obtained in the experimental model of Parkinson's disease provide evidence that the damage of the central dopaminergic pathways induces alterations of some immune functions in mice. PMID- 8423207 TI - Enhancing effects of irrelevant lymphocytes on adoptive transferred experimental allergic encephalomyelitis. AB - To help understand effector mechanisms in experimental allergic encephalomyelitis (EAE) we examined the effects of adding 'irrelevant' lymphocytes from non-EAE donors to major basic protein (MBP)-reactive lymphocytes in the adoptive transfer of EAE (Tr-EAE). The intravenous injection of tetanus toxoid-reactive lymphocytes (TT-cells) and phytohemagglutinin-stimulated lymphocytes on the same day or 2 days after intra-arterial injection of MBP-reactive lymphocytes enhanced the clinical and pathological expression of Tr-EAE. In lymphocyte trafficking studies there was significant accumulation of these injected TT-cells in the central nervous system during enhanced transfer of EAE. W3/25-positive cells were much more predominant in lesion of central nervous system when reinjected with TT cells along with MBP cells compared with lesions of rats injected with MBP cells alone. These findings suggest possible participation of lymphocytes other than MBP-reactive cells in the expression of EAE and provide a useful model to further explore effector mechanisms in the future. PMID- 8423208 TI - Glial cell-specific mechanisms of TGF-beta 1 induction by IL-1 in cerebral cortex. AB - Transforming growth factor beta-1 (TGF-beta 1) immunoreactive product (IRP) has recently been detected in autopsied brains of individuals who died with central nervous system diseases and/or fever but not in normal individuals or in normal rodent brain. However, the mechanism(s) of induction of TGF-beta 1 in brain and the identity of cells expressing TGF-beta 1 need to be understood before a role, if any, for this potent pleiotropic cytokine in neuropathogenesis can be discerned. Towards this end we determined that IL-1 stimulated the production of TGF-beta 1 IRP in cells and TGF-beta 1 activity in culture fluids of all glial cells, astrocytes, microglial cells, and oligodendrocytes, derived from neonatal rat cortex and grown in cell type-enriched cultures. TGF-beta 1 production in vitro varied with the cell type and isoform of IL-1. Oligodendrocytes produced the most and astrocytes the least amount of TGF-beta 1. IL-1 alpha stimulated TGF beta 1 production in all glial cell types, whereas IL-1 beta did not. In vivo, TGF-beta 1 IRP was detected in human tissues from cerebral frontal cortex and subcortical white matter only when interleukin-1 (IL-1) was elevated in the same tissues. Moreover, the amount of detectable TGF-beta 1 was positively correlated with the amount of detectable IL-1 (rho = 0.605; P = 0.003), as determined by morphometry. Double-labelling of cells for their phenotypic markers and expression of TGF-beta 1 indicated that all glial cells, but not neurons, expressed TGF-beta 1. IL-1 alpha and IL-1 beta IRPs were also detected in all three glial cell types, most frequently in astrocytes and least frequently in microglial cells. The cells containing both cytokine IRPs were also detected. These results indicate that TGF-beta 1 may be induced by IL-1 in all glial cells of the frontal cortex, by both autocrine and paracrine mechanisms. PMID- 8423209 TI - Endothelial dysfunction in the coronary microcirculation: a new clinical entity or an experimental finding? PMID- 8423210 TI - Soluble complex of complement increases hydraulic conductivity in single microvessels of rat lung. AB - We determined the effect of sera enriched with the soluble complex of complement (SC5b-9), on hydraulic conductivity (Lp) of single pulmonary venules (diameter 20 30 microns). Sera free of anticoagulants and blood cells were prepared from rat and human blood. Lp were determined by our split drop technique in isolated, blood-perfused lungs prepared from anesthetized rats (2% halothane; Sprague Dawley, 500 g; n = 73). Zymosan-activated (ZAS) and control sera were used for Lp determinations. In ZAS prepared from human serum, SC5b-9 concentration was > 300 micrograms/ml (control: < 1 microgram/ml) as determined by ELISA. At baseline, Lp averaged 3.4 +/- .4 x 10(-7) ml/(cm2.s.cm H2O), but it increased by 217 +/- 32% with undiluted ZAS (P < 0.05). The Lp increase correlated significantly with different ZAS dilutions for rat serum and with SC5b-9 concentration for human serum. Lp did not increase significantly with ZAS prepared from heat-treated sera, C6- and C8-deficient sera; or with ZAS in which SC5b-9 had been depleted by immunoprecipitation. The ZAS-induced increase of Lp was blocked completely by venular preinfusion with the arginine-glycine-aspartic acid (RGD) tripeptide (1 mg/ml, 10 min). We report for the first time that: (a) SC5b-9 increases lung endothelial Lp; and (b) the increase of Lp is attributable to an integrin dependent mechanism. PMID- 8423211 TI - Glucocorticoids stimulate rabbit proximal convoluted tubule acidification. AB - Glucocorticoids have an important role in renal acidification; however, a direct effect of glucocorticoids on proximal convoluted tubule (PCT) acidification has not been directly demonstrated. In the present in vitro microperfusion study PCT from animals receiving dexamethasone (600 micrograms/kg twice daily for 2 d and 2 h before killing) had a significantly higher rate of bicarbonate absorption than did controls (92.0 +/- 13.3 vs 59.9 +/- 3.2 pmol/mm.min, P < 0.01). To examine if glucocorticoids had a direct epithelial action, dexamethasone was added to the bath of PCT perfused in vitro. After 3 h of incubation in paired experiments 10( 6) M and 10(-5) M dexamethasone resulted in an approximately 30% stimulation in the rate of bicarbonate absorption. 10(-7) M dexamethasone and 10(-6) M aldosterone had no effect on bicarbonate absorption. The stimulation of acidification by 10(-5) M dexamethasone was blocked by actinomycin D and cycloheximide. These data are consistent with a direct effect of glucocorticoids on PCT acidification, and this effect is dependent upon protein synthesis. PMID- 8423212 TI - Specificity of T cells invading the skin during acute graft-vs.-host disease after semiallogeneic bone marrow transplantation. AB - The mechanisms responsible for skin lesions during acute graft-vs.-host disease (aGVHD) after allogeneic bone marrow transplantation (BMT) are poorly understood. The exact role of various effector cell populations and "major" (particularly HLA DP) or "minor" antigens as target molecules is not known. To investigate the nature of cells responsible for tissue injury, we cultured T cells from skin biopsy first with interleukin 2 (IL-2) alone and then in polyclonal activation conditions to avoid in vitro antigenic sensitization before specificity testing. We applied this method to two biopsies performed during aGVHD after semiallogeneic BMT and obtained cytotoxic T cells against four graft mismatches: CD8+ T cells against HLA-A2.2 and HLA-B27 and CD4+ T cells against HLA-DP101 and HLA-DP401. This demonstrates that T cells with documented specificity can be obtained from an aGVHD lesion without antigenic selection. Moreover, these data directly implicate DP as a potential target antigen for aGVHD. PMID- 8423213 TI - Development and transfer of immediate cutaneous hypersensitivity in mice exposed to aerosolized antigen. AB - We previously showed that BALB/c mice sensitized to ovalbumin (OVA) by brief daily inhalations of antigen over 10 consecutive days exhibit elevated antigen specific serum IgE antibody levels and increased airways responsiveness. For the first time, we now show that animals sensitized in this fashion to either OVA or ragweed (RGW) develop immediate hypersensitivity skin test reactions when challenged 2 d after completion of the sensitization protocol. Skin testing, performed by direct assessment of wheal formation after intradermal injection of allergen, was sensitive and specific, since animals exposed to RGW by inhalation only responded to RGW, and OVA-sensitized animals responded only to OVA. Positive reactions were associated with mast cell degranulation, whereas control injections were not. Since only sensitized IgE high responder BALB/c mice but neither nonsensitized BALB/c mice nor OVA-sensitized IgE low responder SJL/J mice exhibited wheal responses, induction of OVA-specific IgE appeared to be essential for the mediation of OVA-specific immediate hypersensitivity reactions of the skin in this model. Passive cutaneous anaphylaxis (PCA) testing confirmed the presence of antigen-specific IgE in the serum. Mice that developed IgG (predominantly IgG2b) anti-OVA antibodies did not respond to OVA injection, indicating that OVA-specific IgG was not involved in this system. Further support for the role of IgE in the immediate hypersensitivity response included the wheal response to intradermal injection of anti-IgE antibody that occurred in OVA- and RGW-sensitized mice at 10-fold lower concentrations than in nonsensitized BALB/c mice and not in sensitized SJL/J mice. After transfer of mononuclear cells from peribronchial lymph nodes of OVA- or RGW-sensitized BALB/c mice, naive, syngeneic recipients developed antigen-specific IgE and specific immediate hypersensitivity responses, indicating that the local lymphoid tissue at the site of sensitization can transfer responsiveness to these allergens. These results demonstrate for the first time the ability to elicit and study IgE-mediated immediate skin hypersensitivity responses in the mouse and illustrate the association of increased antigen-specific and total serum IgE levels, airways hyperresponsiveness, and antigen-specific immediate cutaneous reactivity after sensitization to allergen via the airways. PMID- 8423214 TI - Effect of insulin-like growth factor-1 on the responses to and recognition of hypoglycemia in humans. A comparison with insulin. AB - Recombinant human insulin-like growth factor-1 (rhIGF-1) lowers blood glucose in humans but its effect on counterregulatory responses has not been established. We therefore compared infusions of rhIGF-1 (0.7 micrograms/kg per min) and insulin (0.8 mU/kg.min) for 120 min in 10 healthy volunteers (glucose allowed to fall freely). With both, glucose fell rapidly because of stimulation of glucose uptake and suppression of hepatic glucose production. Despite similar plasma glucose nadirs (2.6 +/- 0.1 vs. 2.7 +/- 0.1 mM), the glucagon response was absent (P < 0.005), growth hormone release was attenuated (P < 0.03), and norepinephrine levels were increased (P < 0.05) by rhIGF-1 compared with insulin. Absent glucagon responses were associated with a blunting of the rebound increase in glucose production (P < 0.05 vs. insulin). After stopping the infusions, glucose recovery was delayed with rhIGF-1 (P < 0.001 vs. insulin). To further evaluate the effects of rhIGF-1 during a standard hypoglycemic stimulus, eight additional healthy subjects received rhIGF-1 or insulin while glucose was clamped at 2.8 mM. Again the rise in glucagon during insulin-induced hypoglycemia was totally abolished by rhIGF-1. Growth hormone responses were delayed, whereas increases in norepinephrine, heart rate, and symptomatic awareness of hypoglycemia were greater with rhIGF-1 compared with insulin (P < 0.05). It was concluded that rhIGF-1 suppression of glucagon release during hypoglycemia impairs glucose recovery. Paradoxically, awareness of hypoglycemia is enhanced with rhIGF-1 in part due to stimulation of the sympathetic activity. PMID- 8423215 TI - Expression of vascular endothelial growth factor does not promote transformation but confers a growth advantage in vivo to Chinese hamster ovary cells. AB - Vascular endothelial growth factor (VEGF) is a mitogen with a specificity for endothelial cells in vitro and an angiogenic inducer in vivo. We tested the hypothesis that VEGF may confer on expressing cells a growth advantage in vivo. Dihydrofolatereductase--Chinese hamster ovary cells were transfected with expression vectors which direct the constitutive synthesis of VEGF. Neither the expression nor the exogenous administration of VEGF stimulated anchorage dependent or anchorage-independent growth of Chinese hamster ovary cells in vitro. However, VEGF-expressing clones, unlike control cells, demonstrated an ability to proliferate in nude mice. Histologic examination revealed that the proliferative lesions were compact, well vascularized, and nonedematous. Ultrastructural analysis revealed that capillaries within the lesions were of the continuous type. These findings indicate that the expression of VEGF may confer on cells the ability to grow in vivo in the absence of transformation by purely paracrine mechanisms. Since VEGF is a widely distributed protein, this property may have relevance for a variety of physiological and pathological proliferative processes. PMID- 8423216 TI - High prevalence of mutations of the p53 gene in poorly differentiated human thyroid carcinomas. AB - The development and progression of thyroid tumors is signaled by phenotype specific mutations of genes involved in growth control. Molecular events associated with undifferentiated thyroid cancer are not known. We examined normal, benign, and malignant thyroid tissue for structural abnormalities of the p53 tumor suppressor gene. Mutations were detected by single-strand conformation polymorphisms of PCR-amplified DNA, using primers bracketing the known hot spots on either exons 5, 6, 7, or 8. The prevalence of mutations was as follows: normal thyroid 0/6; follicular adenomas 0/31; papillary carcinomas 0/37; medullary carcinomas 0/2; follicular carcinomas 1/11; anaplastic carcinomas 5/6; thyroid carcinoma cell lines 3/4. Positive cases were confirmed by direct sequencing of the PCR products. All five anaplastic carcinoma tissues and the anaplastic carcinoma cell line ARO had G:C to A:T transitions leading to an Arg to His substitution at codon 273. In both tumors and cell lines, examples of heterozygous and homozygous p53 mutations were identified. The only thyroid carcinoma cell line in which p53 mutations were not detected in exons 5-8 had markedly decreased p53 mRNA levels, suggesting the presence of a structural abnormality of either p53 itself or of some factor controlling its expression. The presence of p53 mutations almost exclusively in poorly differentiated thyroid tumors and thyroid cancer cell lines suggests that inactivation of p53 may confer these neoplasms with aggressive properties, and further loss of differentiated function. PMID- 8423217 TI - Synthesis of oxytocin in amnion, chorion, and decidua may influence the timing of human parturition. AB - Despite the widespread clinical use of oxytocin (OT) as a potent and specific stimulant of labor, previous research data have not supported a role for OT in the physiology of normal human parturition. We have demonstrated synthesis of OT mRNA in amnion, chorion, and decidua using Northern blot analysis, ribonuclease protection assays, and in situ hybridization. Probes directed towards both the 3' and 5' ends of the gene have been used. Levels were highest in decidua with considerably less in chorion and amnion and very low levels in placenta. The transcript size in decidua appears to be 60-80 nucleotides smaller than the transcripts in amnion and chorion. OT gene expression in chorio-decidual tissues increased three- to fourfold around the time of labor onset. Estradiol stimulated synthesis of OT mRNA during in vitro incubation. These results support the hypothesis of a paracrine system involving OT and sex steroids within intrauterine tissues wherein significant changes could occur without being reflected in the maternal circulation. Such a paracrine system could rationalize a long-sought role for oxytocin in the physiology of human labor. These data may lead to novel approaches towards prevention or treatment or preterm labor. PMID- 8423218 TI - Kistrin, an integrin antagonist, blocks endocytosis of fibrinogen into guinea pig megakaryocyte and platelet alpha-granules. AB - Recent data indicate that megakaryocyte/platelet alpha-granule fibrinogen is endocytosed from plasma. Because fibrinogen is the major platelet protein present in high concentrations in alpha-granules, fibrinogen uptake into alpha-granules may occur via specific receptors. In that cells of the megakaryocyte/platelet lineage contain two integrins--alpha IIb beta 3 (GP IIb-IIIa) and the vitronectin receptor (alpha v beta 3)--that can bind fibrinogen, one or both of these receptors may mediate the endocytic uptake of fibrinogen. To test this hypothesis, we examined the effect of Kistrin, an RGD-containing protein purified from the venom of Agkistrodon rhodostoma that inhibits fibrinogen binding to human platelet receptors, on endocytosis of fibrinogen by megakaryocytes and platelets. Continuous intravenous infusion of kistrin into guinea pigs (200 micrograms/h) over a 24-h period inhibited collagen-induced platelet aggregation. When biotinylated fibrinogen was injected intravenously into animals receiving Kistrin, megakaryocytes failed to endocytose the labeled fibrinogen. Endocytosis of fibrinogen into platelets was also inhibited in these animals. In contrast, platelets and megakaryocytes obtained from sham-infused control animals contained the injected biotinylated fibrinogen. We conclude that, in addition to the well known extracellular function of cell adhesion, integrins can also act as receptors that mediate endocytosis of exogenous proteins and incorporate them into regulated secretory granules. PMID- 8423219 TI - Band 3 and glycophorin are progressively aggregated in density-fractionated sickle and normal red blood cells. Evidence from rotational and lateral mobility studies. AB - Band 3 aggregation in the plane of the red blood cell (RBC) membrane is postulated to be important in the pathophysiology of hemolysis of dense sickle and normal RBCs. We used the fluorescence photobleaching recovery and polarized fluorescence depletion techniques to measure the lateral and rotational mobility of band 3, glycophorins, and phospholipid analogues in membranes of density separated intact RBCs from seven patients with sickle cell disease and eight normal controls. The fractions of laterally mobile band 3 and glycophorin decreased progressively as sickle RBC density increased. Normal RBCs also showed a progressive decrease in band 3 fractional mobility with increasing buoyant density. Rapidly rotating, slowly rotating, and rotationally immobile forms of band 3 were observed in both sickle and normal RBC membranes. The fraction of rapidly rotating band 3 progressively decreased and the fraction of rotationally immobile band 3 progressively increased with increasing sickle RBC density. Changes in the fraction of rotationally immobile band 3 were not reversible upon hypotonic swelling of dense sickle RBCs, and normal RBCs osmotically shrunken in sucrose buffers failed to manifest band 3 immobilization at median cell hemoglobin concentration values characteristic of dense sickle RBCs. We conclude that dense sickle and normal RBCs acquire irreversible membrane abnormalities that cause transmembrane protein immobilization and band 3 aggregation. Band 3 aggregates could serve as cell surface sites of autologous antibody binding and thereby lead to removal of dense sickle and normal (senescent) RBCs from the circulation. PMID- 8423220 TI - The antimalarial action of desferal involves a direct access route to erythrocytic (Plasmodium falciparum) parasites. AB - We designed the N-methylanthranilic-desferrioxamine (MA-DFO) as a fluorescent iron (III) chelator with improved membrane permeation properties. Upon binding of iron (III), MA-DFO fluorescence is quenched, thus allowing traceability of drug iron (III) interactions. MA-DFO is well tolerated by mammalian cells in culture. Its antimalarial activity is pronounced: IC50 values on in vitro (24-h) growth of Plasmodium falciparum were 3 +/- 1 microM for MA-DFO compared with 30 +/- 8 for DFO. The onset of growth inhibition of rings or trophozoites occurs 2-4 h after exposure to 13 microM MA-DFO. This effect is commensurate with MA-DFO permeation into infected cells. In a 24-h exposure to MA-DFO or DFO, trophozoites take up either compound to approximately 10% of the external concentration, rings to 5%, and noninfected cells to < 1%. Red cells encapsulated with millimolar concentrations of DFO or MA-DFO fully support parasite invasion and growth. We conclude that extracellular MA-DFO and DFO gain selective access into parasites by bypassing the host. The rate-limiting step is permeation through the parasite membrane, which MA-DFO accomplishes faster than DFO, in accordance with its higher hydrophobicity. These views are consistent with the proposed duct, which apparently provides parasitized cells with a window to the external medium. PMID- 8423221 TI - Diversity of airway epithelial cell targets for in vivo recombinant adenovirus mediated gene transfer. AB - A variety of pulmonary disorders, including cystic fibrosis, are potentially amenable to treatment in which a therapeutic gene is directly transferred to the bronchial epithelium. This is difficult to accomplish because the majority of airway epithelial cells replicate slowly and/or are terminally differentiated. Adenovirus vectors may circumvent this problem, since they do not require target cell proliferation to express exogenous genes. To evaluate the diversity of airway epithelial cell targets for in vivo adenovirus-directed gene transfer, a replication deficient recombinant adenovirus containing the Escherichia coli lacZ (beta-galactosidase [beta-gal]) gene (Ad.RSV beta gal) was used to infect lungs of cotton rats. In contrast to uninfected animals, intratracheal Ad.RSV beta gal administration resulted in beta-gal activity in lung lysate and cytochemical staining in all cell types forming the airway epithelium. The expression of the exogenous gene was dose-dependent, and the distribution of the beta-gal positive airway epithelial cells in Ad.RSV beta gal-infected animals was similar to the normal cell differential of the control animals. Thus, a replication deficient recombinant adenovirus can transfer an exogenous gene to all major categories of airway epithelial cells in vivo, suggesting that adenovirus vectors may be an efficient strategy for in vivo gene transfer in airway disorders such as cystic fibrosis. PMID- 8423222 TI - Increased plasma viscosity as a reason for inappropriate erythropoietin formation. AB - The aim of this study was to examine whether altered plasma viscosity could contribute to the inappropriately low production rate of erythropoietin (EPO) observed in patients suffering from hypergammaglobulinemias associated with multiple myeloma or Waldenstrom's disease. We found that the EPO formation in response to anemia in these patients was inversely related to plasma viscosity. A similar inverse relationship between plasma viscosity and EPO production was seen in rats in which EPO formation had been stimulated by exchange transfusion and the plasma viscosity of which was thereby altered by using exchange solutions of different composition to alter plasma viscosity and thus whole blood viscosity independently from hematocrit. Raising the gammaglobulin concentration to approximately 40 mg/ml plasma in the rats almost totally blunted the rise in serum EPO levels despite a fall of the hematocrit to 20%. Determination of renal EPO mRNA levels by RNase protection revealed that the reductions in serum EPO levels at higher plasma viscosities were paralleled by reductions in renal EPO mRNA levels. Taken together, our findings suggest that plasma viscosity may be a significant inhibitory modulator of anemia-induced EPO formation. The increased plasma viscosity in patients with hypergammaglobulinemias may therefore contribute to the inappropriate EPO production, which is a major reason for the anemia developing in these patients. PMID- 8423223 TI - Macrophage colony-stimulating factor is indispensable for both proliferation and differentiation of osteoclast progenitors. AB - The mechanism of action of macrophage colony-stimulating factor (M-CSF) in osteoclast development was examined in a co-culture system of mouse osteoblastic cells and spleen cells. In this co-culture, osteoclast-like multinucleated cells (MNCs) were formed within 6 d in response to 10 nM 1 alpha,25(OH)2D3 added only for the final 2 d of culture. Simultaneously adding hydroxyurea for the final 2 d completely inhibited proliferation of cultured cells without affecting 1 alpha,25(OH)2D3-stimulated MNC formation. Autoradiographic examination using [3H] thymidine revealed that osteoclast progenitors primarily proliferated during the first 4 d, whereas their differentiation into MNCs occurred predominantly during the final 2 d of culture in response to 1 alpha,25(OH)2D3. When anti-M-CSF antibody or anti-M-CSF receptor antibody was added either for the first 4 d or for the final 2 d, the MNC formation was similarly inhibited. In co-cultures of normal spleen cells and osteoblastic cells obtained from op/op mice, which cannot produce functionally active M-CSF, the lack of M-CSF either for the first 4 d or for the final 2 d failed to form MNCs in response to 1 alpha,25(OH)2D3 added for the last 2 d. These results clearly indicate that M-CSF is indispensable for both proliferation of osteoclast progenitors and their differentiation into mature osteoclasts. PMID- 8423224 TI - A monoclonal antibody recognizes a von Willebrand factor domain within the amino terminal portion of the subunit that modulates the function of the glycoprotein IB- and IIB/IIIA-binding domains. AB - We developed a monoclonal antibody, 1C1E7, against vWf that increases ristocetin induced platelet aggregation in a dose-dependent manner and lowers the threshold concentration of ristocetin needed to obtain a full aggregatory response. The platelet aggregatory effect of asialo vWf (ASvWf) also is enhanced by 1C1E7, in the presence or absence of glycoprotein (GP) IIb/IIIa receptor antagonism. In the presence of ristocetin, both intact 1C1E7 and its Fab fragments enhance specific binding of 125I-vWf to platelets. With 1C1E7, the intermediate and higher molecular weight multimers of vWf are preferentially bound to both GP Ib and GP IIb/IIIa. Thrombin-induced 125I-vWf binding to GP IIb/IIIa also is increased by 1C1E7. Maximal binding of 1C1E7 to vWf corresponds to 0.97 mol/mol vWf monomer with a Kd of 4.7 x 10(-10) M. 1C1E7 reacts with a 34/36-kD tryptic fragment (III T4) and a 34-kD plasmic fragment (P34), which localizes the epitope between amino acid residues 1 and 272; this was confirmed by NH2-terminal amino acid sequencing. Finally, platelet aggregation by ASvWf was associated with a sharp rise in intracellular Ca2+ only in the presence of 1C1E7. An antibody-mediated conformational change of vWf may result in an improved presentation of the GP Ib- and GP IIb/IIIa-binding domains of mainly the larger multimers; the increased density of vWf on the platelet surface leads to platelet activation. The antibody may thus recognize a domain of relevance for vWf physiology. PMID- 8423225 TI - Rhesus monkey lipoprotein(a) binds to lysine Sepharose and U937 monocytoid cells less efficiently than human lipoprotein(a). Evidence for the dominant role of kringle 4(37). AB - Rhesus lipoprotein(a) (Lp[a]) binds less efficiently than human Lp(a) to lysine Sepharose and to cultured U937 cells. Studies using elastase-derived plasminogen fragments indicated that neither kringle 5 nor the protease domain of Lp(a) are required in these interactions pointing at an involvement of the K4 region. Comparative structural analyses of both the human and simian apo(a) K4 domain, together with molecular modeling studies, supported the conclusion that K4(37) plays a dominant role in the lysine binding function of apo(a) and that the presence of arginine 72 rather than tryptophan in this kringle can account for the functional deficiency observed with rhesus Lp(a). These in vitro results suggest that rhesus Lp(a) may be less thrombogenic than human Lp(a). PMID- 8423226 TI - Impaired coronary blood flow response to acetylcholine in patients with coronary risk factors and proximal atherosclerotic lesions. AB - We examined whether coronary risk factors and atherosclerotic lesions in the study artery were associated with impaired endothelium-dependent dilation of coronary resistance arteries. Acetylcholine (ACH) at graded doses (1, 3, 10 and 30 micrograms/min) and papaverine (10 mg) were selectively infused into the left anterior descending coronary artery of 28 patients, in whom the study artery was angiographically normal (n = 16) or with mild stenosis < or = 40% (n = 12). Coronary blood flow (CBF) was estimated from the product of mean CBF velocity measured by an intracoronary Doppler catheter and the arterial cross-sectional area of the study artery determined by quantitative arteriography. ACH increased CBF in a dose-dependent manner. However, the maximum CBF response to ACH varied widely among patients (from 50% to 660%). By multivariate analysis, the presence of atherosclerotic lesions in the study artery was an independent predictor for impaired CBF response to ACH (P < 0.01). Hypertension (P < 0.001), hypercholesterolemia (r = -0.52, P < 0.005), age > or = 50 yr (P < 0.01) and total number of coronary risk factors (r = -0.62, P < 0.001) were associated with the impaired increase in CBF with ACH by univariate analysis. The percent increase in CBF evoked with papaverine did not correlate with these risk factors. The results suggest that mild atherosclerotic lesions in the study artery and coronary risk factors are accompanied by impaired endothelium-dependent dilation of coronary resistance arteries evoked with ACH. Endothelial dysfunction of coronary resistance arteries may result in altered regulation of myocardial perfusion in patients with mild coronary atherosclerosis and coronary risk factors. PMID- 8423227 TI - Clonal expansion of lung V delta 1+ T cells in pulmonary sarcoidosis. AB - Sarcoidosis is a multisystem disease of unknown etiology characterized by the presence of noncaseating granulomas in involved tissues. To investigate a potential role for gamma/delta T cells in the pathogenesis of pulmonary sarcoidosis, we studied lung and blood T cells from patients for preferential expression of particular gamma/delta T cell receptors. An abnormally high percentage of gamma/delta cells was found in the blood of some patients. However, the increased percentage did not reflect an increase in absolute number, and appeared to be secondary to a decrease in T cells expressing alpha/beta receptors. Furthermore, as in normals, the circulating gamma/delta cells in patients predominantly expressed V gamma 9/V delta 2 receptors, a subset that was not enriched at the site of disease. In contrast, in the lung, an increased percentage of gamma/delta cells expressing V delta 1 was found in a subset of patients. Importantly, these cells demonstrated evidence of prior activation by selectively expanding in vitro in the presence of interleukin 2. Furthermore, an analysis of junctional region sequences revealed their clonal nature. These clonal expansions of V delta 1+ cells in pulmonary sarcoidosis provide evidence for a disease process that involves specific recognition of a local antigen by T cells, and contributes new information regarding the nature of the as yet undefined antigenic stimulus. PMID- 8423228 TI - Preserved incretin activity of glucagon-like peptide 1 [7-36 amide] but not of synthetic human gastric inhibitory polypeptide in patients with type-2 diabetes mellitus. AB - In type-2 diabetes, the overall incretin effect is reduced. The present investigation was designed to compare insulinotropic actions of exogenous incretin hormones (gastric inhibitory peptide [GIP] and glucagon-like peptide 1 [GLP-1] [7-36 amide]) in nine type-2 diabetic patients (fasting plasma glucose 7.8 mmol/liter; hemoglobin A1c 6.3 +/- 0.6%) and in nine age- and weight-matched normal subjects. Synthetic human GIP (0.8 and 2.4 pmol/kg.min over 1 h each), GLP 1 [7-36 amide] (0.4 and 1.2 pmol/kg.min over 1 h each), and placebo were administered under hyperglycemic clamp conditions (8.75 mmol/liter) in separate experiments. Plasma GIP and GLP-1 [7-36 amide] concentrations (radioimmunoassay) were comparable to those after oral glucose with the low, and clearly supraphysiological with the high infusion rates. Both GIP and GLP-1 [7-36 amide] dose-dependently augmented insulin secretion (insulin, C-peptide) in both groups (P < 0.05). With GIP, the maximum effect in type-2 diabetic patients was significantly lower (by 54%; P < 0.05) than in normal subjects. With GLP-1 [7-36 amide] type-2 diabetic patients reached 71% of the increments in C-peptide of normal subjects (difference not significant). Glucagon was lowered during hyperglycemic clamps in normal subjects, but not in type-2 diabetic patients, and further by GLP-1 [7-36 amide] in both groups (P < 0.05), but not by GIP. In conclusion, in mild type-2 diabetes, GLP-1 [7-36 amide], in contrast to GIP, retains much of its insulinotropic activity. It also lowers glucagon concentrations. PMID- 8423229 TI - Differentiation-associated switches in protein 4.1 expression. Synthesis of multiple structural isoforms during normal human erythropoiesis. AB - Erythroid differentiation is accompanied by dramatic alterations in morphology and membrane mechanical properties resulting, in large part, from reorganization of the membrane skeletal protein network. The 80-kD protein 4.1 is an important organizational component of this membrane skeleton. Recently, it has been recognized that multiple structural isoforms of 4.1 are encoded by a single gene via alternative pre-mRNA splicing, and that an upstream ATG can be spliced in and used for translation of high molecular weight 4.1. We are exploring the hypothesis that differentiation-associated switches in protein 4.1 structure play an important role in membrane reorganization. To study changes in 4.1 gene expression during normal human differentiation, we analyzed 4.1 protein and mRNA structure at various developmental stages. Using immunofluorescence microscopy, we observed high molecular weight 4.1 isoforms in preproerythroblasts producing punctate, predominantly cytoplasmic staining with a perinuclear area of intense fluorescence, while mature red cells expressed very little high molecular weight 4.1. Isoforms containing an alternatively expressed 102-nucleotide exon near the COOH terminus were abundant in both preproerythroblasts and mature cells but produced a punctate distribution of fluorescence over the entire preproerythroblast and intense membrane-associated fluorescence in the erythrocyte. Characterization of RNA by polymerase chain reaction and nuclease protection assays revealed a differentiation-associated switch in pre-mRNA splicing in the spectrin-actin binding domain. Since this domain plays a critical role in regulating membrane material properties, we speculate that this switch may be crucial to reorganization of the skeletal network during erythropoiesis. We conclude that 4.1 isoforms are differentially expressed and differentially localized during erythropoiesis, and that this isoform family is likely to have diverse functions during terminal differentiation. PMID- 8423230 TI - Antibody to Rmp (outer membrane protein 3) increases susceptibility to gonococcal infection. AB - The severe adverse effects of gonococcal infection on human fertility suggests that Neisseria gonorrhoeae would exert powerful selection for the development of a protective immune response in humans. N. gonorrhoeae is an obligate human pathogen and must persist in humans to survive. Since it is an ecologically successful organism, it must have evolved strategies to evade any human immune response it elicits. In a longitudinal study among 243 women working as prostitutes and experiencing frequent gonococcal infection, younger women, women with HIV infection, and women with antibody to the gonococcal outer membrane protein 3 (Rmp) were at increased risk of infection (adjusted odds ratio 3.4, CI95% 1.1-10.4, P < 0.05). Rmp is highly conserved in N. gonorrhoeae and the blocking of mucosal defences may be one of its functions. As similar proteins occur in many gram negative mucosal pathogens, the enhancing effect of such proteins may be a general strategy whereby bacteria evade human immune responses. PMID- 8423231 TI - Islet cell cytoplasmic autoantibody reactivity to glutamate decarboxylase in insulin-dependent diabetes. AB - Individuals with or at risk for insulin-dependent diabetes (IDD) frequently have autoantibodies against an islet cell cytoplasmic (ICA) antigen thought to be a sialoglycolipid. However, we now report that preabsorption of ICA-positive sera with recombinant glutamate decarboxylase (human GAD 65 and/or GAD 67) reduced or blocked the ICA reactivity of 5/18 (27%) new-onset IDD patients and 7/18 (39%) prediabetics. Interestingly, nondiabetic subjects with ICA of > or = 5 yr in duration had GAD-reactive ICA significantly more often (16/24, 67%, P < 0.04) than the diabetic groups. ICA reactivity to GAD was not related to serum ICA titer nor the age of the individual, and in all cases tested was blocked by GAD 65 or GAD 67 with equivalent efficiency. The ICA observed in 21/25 (84%) IDD patients with ICA long after clinical onset of disease (9-42 yr) was reactive to GAD. A natural history analysis of three individuals showed conversions from ICA which was reactive to GAD to a non-GAD-reactive ICA nearer to their clinical onsets of IDD. This study further defines the autoantigens reactive to ICA, and suggests that, whereas ICA that are not reactive to GAD may identify an advanced and more prognostic lesion, GAD-reactive ICA may typify the early or inductive lesion that may or may not progress to clinically significant beta cell injury. PMID- 8423232 TI - Quantitative assay using recombinant human islet glutamic acid decarboxylase (GAD65) shows that 64K autoantibody positivity at onset predicts diabetes type. AB - At and before onset, most insulin-dependent diabetics (IDDM) have islet GAD65 autoantibodies (GAD65Ab). Since IDDM also occurs in older patients where non insulin-dependent diabetes is common, we studied GAD65Ab at onset to classify diabetes type. Our quantitative immunoprecipitation assay uses recombinant human islet GAD65 stably expressed in hamster fibroblasts. Electrophoretic mobility was identical to native islet GAD65. Like native antigen, recombinant GAD65 migrated as two bands during electrophoresis, but converted to one under stronger reduction. Immunoprecipitation was linear with respect to antibody or antigen concentration. In 120 population-based diabetic patients of all ages grouped by treatment at onset and after 18 mo, GAD65Ab were present in 70% on insulin (n = 37), 10% on oral agent (n = 62, P < 0.0001), 69% changing from oral agent to insulin (n = 16, P < 0.001), and 1 of 33 controls. 65% with GAD65Ab, versus 8% without, changed from oral agent to insulin (P < 0.01). The GAD65Ab quantitative index was remarkably stable, and only 2 of 32 patients changed antibody status during follow-up. Concordance between GAD65Ab and islet cell antibodies was 93%. Quantitative correlation was approximate but significant. This highly sensitive, quantitative, high capacity assay for GAD65Ab reveals treatment requirements better than clinical criteria, perhaps guiding immunomodulatory therapy. PMID- 8423233 TI - Serum-derived growth factor is thrombin? PMID- 8423234 TI - Characterization of the internalization of bacillus Calmette-Guerin by human bladder tumor cells. AB - Adjuvant intravesical Mycobacterium bovis BCG is the treatment of choice for recurrent superficial bladder cancer. Fibronectin (FN) was previously demonstrated to be necessary for the retention of BCG within the bladder and for the expression of antitumor activity. Recent studies have demonstrated that BCG attach and are ingested by bladder epithelial cells, suggesting the existence of a second bacterial attachment mechanism. We report the characterization of the molecules involved in BCG attachment and internalization by the human bladder transitional cell carcinoma cell line T-24. Pretreatment of T-24 cells with monoclonal antibodies to either alpha 5 or beta 1 integrin subunits significantly inhibited both BCG attachment and ingestion. Exogenous FN was observed to enhance both attachment and ingestion of BCG, and anti-FN was observed to inhibit both phenomena. Latex beads precoated with either FN or laminin (LN) but not BSA were ingested by T-24 cells, but only FN-coated beads inhibited BCG attachment and ingestion. Pretreatment of BCG with FN augmented both attachment and ingestion. The role of bacterial FN binding proteins was evaluated. A monoclonal antibody to a 55-kD FN-binding protein was observed to abrogate attachment and ingestion. These results demonstrate that attachment and ingestion of BCG are mediated in part by the alpha 5 beta 1 integrin receptor and are dependent on FN. These studies demonstrate a mechanism of entrance of mycobacteria into epithelial cells and suggest a second role for FN in the adjuvant antitumor effect of BCG. PMID- 8423235 TI - An isoform-specific mutation in the protein 4.1 gene results in hereditary elliptocytosis and complete deficiency of protein 4.1 in erythrocytes but not in nonerythroid cells. AB - Multiple protein 4.1 isoforms are expressed in a variety of tissues through complex alternative pre-mRNA splicing events, one function of which is to regulate use of two alternative translation initiation signals. Late erythroid cells express mainly the downstream initiation site for synthesis of prototypical 80-kD isoforms; nonerythroid cells in addition use an upstream site to encode higher molecular mass isoform(s). In this study, we examined the effects of a 5' gene rearrangement in a family with hereditary elliptocytosis and complete deficiency of erythrocyte 4.1 protein on 4.1 isoform expression in erythroid vs. nonerythroid cells. Patient 4.1 mRNAs from reticulocytes, fibroblasts, and B lymphocytes were amplified by reverse transcriptase/polymerase chain reaction techniques and shown to exhibit a 318-nucleotide deletion that encompasses the downstream AUG, but leaves intact the upstream AUG. Immunoblot analysis revealed a total deficiency of 4.1 in patient red cells and a selective deficiency of 80 kD isoform(s) but not high molecular weight 4.1 in patient nonerythroid cells. Thus, the 4.1 gene mutation in this family produces an isoform-specific deficiency that is manifested clinically in tissue-specific fashion, such that red cells are affected but other cell types are unaffected because of tissue specific differences in RNA splicing and translation initiation. PMID- 8423236 TI - Lovastatin inhibits proliferation of rat mesangial cells. AB - Products of intracellular mevalonate metabolism are essential for cell growth and proliferation. Lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, blocks the formation of mevalonate and its metabolites, and has been shown to inhibit proliferation of several cell types. In vivo, lovastatin has reduced mesangial cellularity and glomerular injury in experimental renal disease. In this study, we investigated the effects of lovastatin on DNA replication and proliferation in rat glomerular mesangial cells. Growth-arrested mesangial cells were exposed to medium containing 10% fetal bovine serum to stimulate mitogenesis. Lovastatin (1-20 microM) caused a significant (P < 0.05) dose-dependent reduction in DNA synthesis ([3H]thymidine incorporation) which was completely prevented in the presence of exogenous mevalonate (100 microM). Lovastatin (1 microM) inhibited cell proliferation by 90% over a 5-d period, and this was largely overcome by added mevalonate. Exogenous low density lipoprotein (100 micrograms/ml) did not prevent lovastatin inhibition of DNA synthesis. The isoprenoid end product isopentenyl adenine (5 or 50 microM) had little effect on DNA synthesis and cell proliferation in lovastatin-blocked cells. By contrast, the isoprenoid farnesol (5 microM) largely prevented lovastatin inhibition of DNA synthesis. We conclude that mevalonate metabolism is essential for mesangial cell proliferation, possibly through the production of the isoprenoid farnesol. Moreover, the action of lovastatin to reduce experimental glomerular injury may involve a direct effect on mesangial cells. PMID- 8423237 TI - Expression of high affinity interleukin-4 receptors on human renal cell carcinoma cells and inhibition of tumor cell growth in vitro by interleukin-4. AB - Previously, Puri et al. (Puri, R. K., M. Ogata, P. Leland, G. M. Feldman, D. Fitzgerald, and I. Pastan. 1991. Cancer Res. 51:3011-3017) have demonstrated that murine sarcoma and colon adenocarcinoma cells express high affinity interleukin-4 receptors (IL-4R) which are internalized after binding to a chimeric ligand consisting of IL-4 and Pseudomonas exotoxin. In the present study, we have tested primary cultures of human renal cell carcinoma (RCC) cells, generated from tumor specimens obtained after nephrectomy, for the expression of IL-4R and their modulation by IL-4. By using iodinated IL-4 in a receptor binding assay, we observed that renal cell carcinoma cells expressed a single class of high affinity IL-4R ranging from 1,425 +/- 207 (mean +/- SEM) to 3,831 +/- 299 (mean +/- SEM) IL-4R molecules/cell with a Kd ranging from 112 +/- 11 pM to 283 +/- 71 pM. Northern blot analysis for IL-4R gene expression, performed with a cDNA probe to IL-4R, revealed that all RCC cells exhibited a single mRNA species of 4 kb. IL 4 downregulated the surface expression of IL-4R on one RCC tumor cell line. The function of IL-4R expression on RCC tumor cells was further determined by investigating the effect of IL-4 on tumor cell growth in vitro and comparing it with IL-4 effect on growth of normal fibroblast and endothelial cell lines. Tumor cell growth, as measured by [3H]thymidine incorporation, was inhibited by IL-4 from 20 to 68% in a dose-dependent manner. A neutralizing antibody to human IL-4 was able to reverse the growth inhibitory effect of IL-4. Normal human fibroblast and endothelial cell lines also expressed high affinity IL-4R, however, IL-4 did not inhibit their growth in vitro. In fact, IL-4 caused modest stimulation of their growth. Taken together, our findings can help develop strategies for the treatment of RCC in which IL-4R may be used as a target for IL-4 itself, for IL-4 toxin therapy or, alternatively, in gene therapy. PMID- 8423238 TI - Carrier detection in families with properdin deficiency by microsatellite haplotyping. AB - Human properdin deficiency is an X-linked disorder strongly predisposing to meningococcal disease which has been recorded in over 50 cases of various ethnic origins. Immunochemically, total deficiency (type I), partial deficiency (type II), and deficiency due to a dysfunctional molecule (type III) can be differentiated. It is therefore most likely that the causative molecular defects will show considerable genetic heterogeneity. Analysis of the properdin locus at Xp11.3-Xp11.23 has led to the characterization of two polymorphic (dC-dA)n.(dG dT)n repeats located approximately 15 kb downstream from the structural gene. Three families (two Scottish Caucasoid, one Tunisian Sephardic) with seven deficient individuals were investigated immunochemically and using a nonradioisotopic polymerase chain reaction-based method for microsatellite detection. Probable and definite carriers frequently showed properdin levels which were in the normal range. No recombinants between the microsatellite loci and properdin deficiency were detected, thus allowing identification of the defective allele through the generations in all three pedigrees. Haplotyping for these highly polymorphic microsatellites in close physical linkage to the properdin gene can provide rapid and nonradioactive detection of carrier status and prenatal diagnosis without extensive sequencing analysis. PMID- 8423239 TI - Pre- and postnatal development of efferent connections of the cochlear nucleus in the rat. AB - Although the connections of the auditory brainstem nuclei are well described in adult mammals, almost nothing is known concerning how and when these connections develop. The purpose of the present study was to describe the development of the efferent projections of the cochlear nucleus (CN), the first central relay station in the ascending auditory pathway of mammals. We used two tracers in rats aged between embryonic day 15 (E15) and postnatal day 14 (P14; birth in the rat is at E22 = P0). The carbocyanine dye DiI was applied into the CN in aldehyde fixed tissue. The second tracer, biocytin, was applied into the ventral acoustic stria in an in vitro slice preparation. The ontogeny of the efferent projections from the CN could be divided into three periods. The first period (E15-E17) is characterized by axonal outgrowth. Axons traverse nuclei in the superior olivary complex and the lateral lemniscus and finally grow up into the inferior colliculus, but axon collaterals do not form during this period. The second period (E18-P5) is marked by pronounced collateral branching of CN fibers in auditory brainstem nuclei. Collateralisation in the contralateral inferior colliculus starts shortly before that in the ipsilateral superior olivary complex. The remaining auditory nuclei become successively innervated, as indicated by collaterals found in them. During the third period (P5-P14) terminal structures mature further, as shown by the morphological changes of the calyces of Held in the medial nucleus of the trapezoid body. In conclusion, our results show that the efferent connections from the cochlear nucleus form over a period of almost two weeks and are laid down without forming aberrant internuclear connections. On a nuclear level, an adult-like projection pattern is already achieved one week prior to the onset of physiological hearing. PMID- 8423240 TI - Development of the mesencephalic nucleus of the trigeminal nerve in chick embryos: target innervation, neurotrophin receptors, and cell death. AB - The goal of this study was to determine whether processes of neurons in the mesencephalic nucleus of the trigeminal nerve (Mes V) of chick embryos arrive in their peripheral target prior to the period of developmental cell death, and to determine whether neurons with early target contact survive to a greater extent than neurons with processes that reach their peripheral target later. The arrival of Mes V nerve fibers in the masticatory muscles was determined by injecting the fluorescent tracer DiI, and the position of labeled and unlabeled neurons was mapped in subdivisions of the Mes V nucleus. Developmental changes in the numerical configuration of Mes V subdivisions were studied in DiI-labeled as well as Nissl-stained material. The expression of low-affinity (p75) neurotrophin receptors was investigated throughout development of the Mes V nucleus with in situ hybridization to assess whether and how levels of expression of this trophic receptor may relate to target innervation and cell death. The extent of cell death was evaluated by counting pyknotic nuclei. Processes of Mes V neurons invade their peripheral target between 5 and 7 days of incubation (E5-7). At E7 12, between 800 and 1,400 labeled Mes V neurons were distributed throughout the two main subdivisions of the Mes V nucleus, the tectal commissure and the optic tectum. Only few Mes V neurons were labeled in the posterior commissure or outside the brain. Cell counts in Nissl-stained material from E7-13 revealed that the numbers of Mes V neurons in the optic tectum decreased to about 40-60%, and in the tectal commissure to 20-25%, whereas Mes V neurons in the posterior commissure disappeared almost entirely. Few Mes V neurons remained in the leptomeninges at E8-10, but a considerable number was found outside the midbrain at E11, indicating ongoing migration of some Mes V neurons. Neurotrophin receptors were differentially expressed in the Mes V nucleus: Before and after the period of cell death, 90-100% of Mes V neurons expressed neurotrophin receptors, whereas during, and immediately preceding the period of developmental cell death (E9-E13), merely 70% of Mes V neurons expressed this receptor. These findings are consistent with the hypothesis that early target contact may provide an advantage for the survival of Mes V neurons and that competition for trophic factors may occur in the peripheral target of this nucleus prior to the period of cell death. PMID- 8423241 TI - Early pattern of neuronal differentiation in the Xenopus embryonic brainstem and spinal cord. AB - Wholemount antibody labeling techniques and horseradish peroxidase backfilling were used to analyze the pattern of neuronal differentiation in the embryonic Xenopus central nervous system between stages 22 and 35/36. In the spinal cord, the first neurons to differentiate are the Rohon-Beard neurons; they are followed by ventral neurons with descending axons (descending interneurons, motoneurons) and lateral interneurons with commissural axons. The somata and axons of these primary neurons form dorsal, ventral, and lateral columns, respectively; the ventral and lateral columns uninterruptedly continue forward into the brainstem. The distribution and projection patterns of spinal neurons were analyzed quantitatively. Rohon-Beard neurons, commissural interneurons, and primary motoneurons vary in number from segment to segment. Thus, these neurons are not distributed in a segmental pattern. In each segment, neurons of a given type project axons whose length varies over a wide range. The numerical distribution of axons formed by a population of neurons of a given type was calculated and expressed as the projection profile of these neurons. For each type of neuron and spinal segment, the projection profile is different. Furthermore, the projection profiles change in a systematic way along the spinal cord. For example, the fraction of Rohon-Beard neurons with long ascending axons steadily increases if one moves towards caudal spinal levels. The findings suggest that suprasegmental cues with a graded distribution along the spinal cord determine the number and projection profile of a particular cell type in a given segment. PMID- 8423242 TI - Sensory innervation in the inner conical body of the vibrissal follicle-sinus complex of the rat. AB - The innervation of the inner conical body of the vibrissal follicle-sinus complex of the rat was examined by high-voltage and conventional transmission electron microscopy of serial and semi-serial sections. The inner conical body is innervated by axons supplied almost exclusively by several superficial vibrissal nerves that arise from the infraorbital branch of the trigeminal nerve and converge upon the neck of the follicle-sinus complex. Each superficial vibrissal nerve contains a few A delta myelinated axons and several bundles of 20-30 unmyelinated axons. These axons enter the inner conical body and distribute circumferentially within 7-10 ring-like arrays that encircle the vibrissal follicle and are stacked through the superficial-to-deep extent of the inner conical body. Each ring consists of 1 or 2 myelinated axons and several small bundles of 2-15 unmyelinated axons enclosed in sheaves of parallel collagen fibrils. Myelinated axons provide exclusively lanceolate endings that may arise at the termination of the axon or at nodes of Ranvier. Within the small bundles, unmyelinated axons individually terminate in succession as abrupt cytoplasmic swellings referred to as cytoplasmic blebs, which contain mitochondria or clusters of clear or dense-core vesicles. Because of their affiliation with collagen fibrils and the proximity of myelinated axons, the blebbed endings may have been misinterpreted as Ruffini endings in previous studies. Their structure, distribution, and origin from unmyelinated axons suggest that the blebbed endings may constitute a unique array of low-threshold C-mechanoreceptors. PMID- 8423243 TI - Spinal neurogenesis and axon projection: a correlative study in the rat. AB - The purpose of the present study was to determine the relationship between the duration of a spinal neuron's neurogenic period and the length of its axon or level of projection. Spinal segment L1 was chosen for examination and neurons were divided into four projection groups: 1) supraspinal projection (SSp), 2) long ascending propriospinal (LAPr), 3) short ascending propriospinal (SAPr), and 4) descending propriospinal (DPr). To determine the duration of the neurogenic period for each group, 3H-thymidine was administered to fetal rats during the proliferative period for spinal neuroblasts on one of embryonic (E) days E13 through E16. Between 50 and 100 days after birth neurons in each group were labeled with the retrograde fluorescent tracer Fluoro-Gold. To demonstrate nerve cells with SSp projections, spinal cords were hemisected at spinal segment C3 in one group of animals and Fluoro-Gold was applied to the sectioned surface of the cord. Three additional sets of animals were used to label nerve cells with LAPr, SAPr, and DPr projections by injecting Fluoro-Gold into the gray matter at spinal segments C6, T12, and L5, respectively. Neurons labeled with both Fluoro-Gold and 3H-thymidine and neurons labeled with Fluoro-Gold alone in each animal in each group were counted and the data statistically analyzed. Results showed that within each spinal lamina neurons with different projections were generated, i.e., completed cell division, at significantly different rates. Neurons with the longest axons, those with SSP projections, were generated first. These were followed by those with LAPr projections, and finally those with SAPr and DPr projections. In most laminate there was no significant difference between the neurogenic periods of rostrally projecting short propriospinal (SAPr) neurons versus caudally projecting short propriospinal (DPr) neurons. It was concluded that the duration of the neurogenic period for a given group of neurons within each spinal lamina is inversely related to the distance between the nerve cell and its projection site regardless of the direction of its projection. PMID- 8423244 TI - Cholinergic projection to the dorsal cap of the inferior olive of the rat, rabbit, and monkey. AB - The inferior olive is divided into several subnuclei that receive specific sensory information. The caudal dorsal cap of the medial accessory subdivision of the inferior olive receives horizontal optokinetic information from the nucleus of the optic tract. The immediately subjacent beta-nucleus receives vertical vestibular information mediated by a GABAergic pathway originating from the ipsilateral descending and medial vestibular nuclei. None of the transmitters to the dorsal cap have been identified. Using choline acetyltransferase (ChAT) immunohistochemistry, we have identified a cholinergic pathway that terminates exclusively in the dorsal cap of rats and monkeys. No other division of the inferior olive received a significant cholinergic innervation. In the rabbit, immunostaining for ChAT reveals a weaker and more diffuse cholinergic innervation of both the dorsal cap and the subjacent beta-nucleus. In rats and rabbits we injected iontophoretically the orthograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) into the medial and descending vestibular nuclei (MVN, DVN) as well as the nucleus prepositus hypoglossi (NPH) in order to trace the possible origin of the cholinergic projection. PHA-L injections into the NPH and medial aspect of the MVN labeled terminals within the contralateral dorsal cap. PHA-L injections in the central and lateral aspects of the MVN as well as the DVN labeled the ipsilateral beta-nucleus. Pressure injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) in the caudal dorsal cap of the rabbit inferior olive demonstrated a predominantly contralateral projection to the dorsal cap from the lateral aspect of the NPH. However, pressure injections of HRP into the caudal dorsal cap combined with ChAT immunohistochemistry in the rabbit demonstrated that most of the neurons of the NPH that projected to the dorsal cap were not cholinergic, and that most of the ChAT-positive neurons within the NPH occupied a more ventral location than the neurons within the NPH that were retrogradely labeled from the HRP injection into the contralateral dorsal cap. In the rat, we made lesions in the MVN, DVN and NPH by injection of ibotenic acid (0.3-0.5 microliter), in an attempt to deplete the dorsal cap of the inferior olive of its cholinergic input. Lesions confined to the NPH and medial aspect of the MVN of the rat caused a loss of ChAT staining in the contralateral dorsal cap. Lesions placed more laterally within the MVN or DVN failed to deplete ChAT-positive terminals in the contralateral or ipsilateral dorsal caps. The dorsal cap of the rat and monkey receives a discrete cholinergic projection.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423245 TI - Pulp capping with Bioglass and autologous demineralized dentin in miniature swine. AB - The purpose of this study was to compare the responses of mechanically exposed dental pulps which had been capped with three dissimilar materials: a bioactive ceramic (Bioglass), autologous demineralized dentin matrix (DDM), and a calcium hydroxide product (Life), with Teflon discs as controls. Mechanical dental pulp exposures were made after preparation of deep buccal Class V cavities in 48 teeth in four miniature swine. The exposures were capped and the cavity preparations restored with zinc oxide-eugenol (IRM) cement. The animals were killed after 90 days, the coronal 2/3 of the teeth removed, and sections prepared for either histological or microradiographic examination. The pulpal inflammatory reactions and the degree of reparative dentin formation were assessed from demineralized serial sections. A qualitative assessment of the degree of mineralization of the reparative dentin was made from microradiographs of undecalcified sections. The observations suggest that reparative dentin formation occurs under a variety of pulp-capping materials, but the structure of the reparative dentin varies with the material and the condition of the underlying pulp. PMID- 8423246 TI - Influence of dentinal fluid and stress on marginal adaptation of resin composites. AB - The influence of dentinal fluid and of a number of stress procedures on the quality of the margins of class V restorations located in both enamel and dentin was quantitatively assessed in vitro with the aid of a scanning electron microscope. The materials tested were GLUMA 2000 experimental, Prisma Universal Bond 3, and Syntac, together with the fine hybrid composites supplied by the respective manufacturers (Pekafill, AP.H, and Tetric). All materials achieved over 95% of "continuous margin" in enamel before and after stressing. In dentin, the initial values, with as well as without dentinal fluid simulation, were situated between 93.2 and 98.2%. With GLUMA 2000 experimental after stressing, a "continuous margin" occurred in only 50.2%, but with Prisma Universal Bond 3 and Syntac, the value was 79.0%. The influence of dentinal fluid simulation was dependent on the dentinal adhesive used. The effects of the various stress procedures were not significantly different. PMID- 8423247 TI - Immunocytochemical localization of cathepsin D in rat junctional epithelium. AB - Localization of cathepsin D was studied in the junctional epithelium (JE) of healthy rat gingivae by immuno-light and -electron microscopy, by means of both the avidin-biotin-peroxidase complex method and a colloidal gold IgG method. At the light-microscopic level, cathepsin D was demonstrated in the JE and oral sulcular epithelium (OSE). Cathepsin D immunoreactivity was remarkable in the coronal portion of the JE and decreased toward its apical portion. However, cathepsin D immunoreactivity in the basal cell layer of the JE was negligible or negative. In the OSE, the granular layer was positive for cathepsin D. In the adjacent connective tissue, many macrophage-like cells (not clear at this level) close to the basal cell layer showed strong immunoreactivity. At the electron microscopic level, cathepsin D was found in the primary lysosomes and trans cisternae of Golgi apparatus in the JE cells. These lysosomes were often fused together or were fused with cathepsin D-negative intracytoplasmic vacuoles to form secondary lysosomes, which indicated that intracellular digestion may have been in progress. However, neutrophils contained few gold particles based on cathepsin D. It is likely that the amounts of cathepsin D contained in the JE cells and macrophages are larger than those of cathepsin D contained in the neutrophils. These findings provided morphological evidence that JE cells have the same endocytotic capacity as macrophages and neutrophils, and that JE cells participate in the intracellular digestion that is carried out by lysosomal enzymes such as cathepsin D. It is suggested, in addition, that maximum intracellular digestion occurs in the coronal portion of the JE. PMID- 8423248 TI - The association of mutans streptococci and non-mutans streptococci capable of acidogenesis at a low pH with dental caries on enamel and root surfaces. AB - Coronal dental plaque from each of 12 caries-positive subjects (Group I) was pooled from "white spot" tooth surface areas and, separately, from sound surface areas; sound surface areas in each of 18 caries-free subjects (Group II) were sampled similarly. Two samples, one consisting of material from a root-surface lesion and another of plaque from a sound root-surface area, were obtained from each of another 10 subjects (Group III). The samples from Groups I and II were evaluated for: (1) pH-lowering potential in vitro with dispersed plaque suspensions, excess glucose supply, and a 60-minute test; (2) the levels of mutans streptococci (MS) and lactobacilli; and (3) the distribution of the predominant non-mutans streptococci (non-MS) according to their final pH in glucose broth; only microbial analysis was done for the Group III samples. The levels of the MS were generally positively associated with caries. A weaker positive association was found for the levels of those non-MS capable of acidogenesis at low pH (final pH < 4.4). The latter generally far outnumbered the MS in all types of samples. The levels of lactobacilli were nearly always very low. The pH-lowering potential (final pH and pH drop rate) was higher for plaque from "white spot" areas than for plaque from sound surface areas (Group I). The samples from caries-free subjects (Group II), however, exhibited a pH-lowering potential which was not significantly different from that of both types of samples from the caries-active subjects (Group I).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423249 TI - In vitro antimicrobial activity of the human neutrophil cytosolic S-100 protein complex, calprotectin, against Capnocytophaga sputigena. AB - Calprotectin is a complex of two anionic proteins found in abundance in the cytosol of neutrophils, certain macrophages, and oral epithelial keratinocytes. Bacteria of the genus Capnocytophaga are pathogens of periodontal origin which can cause systemic infection in neutropenic subjects. Recently, it has been observed that Capnocytophaga may be internalized by neutrophils within the cytosol rather than within a membrane-delimited phagosome. The purpose of this study was to test the in vitro antibacterial effect of the cytosolic complex, calprotectin, against Capnocytophaga sputigena. Calprotectin was purified from the cytosol of human neutrophils by gel filtration and anion exchange FPLC, and it exerted potent in vitro antimicrobial effects against C. sputigena. Net bacteriostatic activity was exerted up to 18 h, after which bactericidal effects were observed. Both net bacteriostatic and bactericidal activity occurred at concentrations above 20 micrograms/mL and exhibited identical dose-response characteristics. Particle counts increased in the presence of calprotectin, despite net bacteriostasis as assessed by changes in colony-forming units (CFU). Dose-response characteristics and direct particle counts suggested that net bacteriostatic effects were the result of balanced cell division and death, rather than suspension of cell division. We conclude that calprotectin can be a significant contributor to host defense against infection by Capnocytophaga. PMID- 8423250 TI - Synergistic, growth-inhibitory effects of chlorhexidine and copper combinations on Streptococcus mutans, Actinomyces viscosus, and Actinomyces naeslundii. AB - The purpose of this investigation was to determine whether chlorhexidine combined with copper would exert a synergistic, growth-inhibitory effect against selected oral bacteria. Actinomyces viscosus, Actinomyces naeslundii, and Streptococcus mutans were all susceptible to chlorhexidine individually, with S. mutans displaying the highest sensitivity. Much higher concentrations of copper were needed to achieve growth inhibition of the micro-organisms tested. Determination of minimum inhibitory concentrations (MICs) of chlorhexidine and Cu2+ combinations suggested synergistic activity. Bactericidal kinetics assays confirmed synergism of chlorhexidine and Cu2+ combinations with 1 to 2 log greater decreases in viable cell numbers compared with chlorhexidine alone. Under the constraints of the conditions employed, these data demonstrate the efficacy of chlorhexidine and Cu2+ combinations against the oral bacteria tested. These studies suggest that a chlorhexidine and copper formulation could be useful as a mouthrinse in helping control cariogenic micro-organisms and/or in the treatment of gingivitis. PMID- 8423251 TI - Caries risk assessment by a cross-sectional discrimination model. AB - Although the prevalence of dental caries is continuing to decline, it still affects a majority of the US population and can be a serious problem for those afflicted. The objective of this project was to develop and perfect a model for assessment of risk of dental caries onset in children. In the first study, reported herein, a set of clinical, microbiological, biochemical, and socio demographic variables was identified that distinguished, with an acceptable level of sensitivity and specificity, between children who had no previous caries experience and children who had high caries levels. A total of 313 children--age 12-15 years, 140 from a fluoridated community and 173 from a fluoride-deficient community--was selected on the basis of previous caries experience, either zero DMFS or high DMFS (> or = 6 in the fluoridated or > or = 8 in the fluoride deficient community). Clinical exams for DMFS, dental fluorosis, and plaque were conducted. Stimulated whole saliva was collected for analysis of mutans streptococci, lactobacilli, total viable flora, and fluoride concentration. A questionnaire was used for collection of demographic data as well as information on prior fluoride exposure, dietary habits, and oral hygiene practices. By means of discriminant analyses, with use of seven key clinical and laboratory variables, it was possible for zero-DMFS subjects to e classified correctly (specificity) in 77.6% of cases in the fluoridated community and in 86.1% of cases in the fluoride-deficient community. High-caries subjects were classified as such (sensitivity) in 79.3% and 88.1% of cases, respectively. PMID- 8423252 TI - Familial aggregation of periodontal indices. AB - Analysis of three measures of periodontal health (plaque index, gingival index, and attachment loss) was carried out on 178 individuals in 75 families examined as part of a family study of periodontal health. Original participants in this study were volunteers recruited from the University of Maryland Dental Clinic, and were selected independently of any specific dental disease or condition. Relatives were invited to participate in the family study so that the extent of familial aggregation of indices of periodontal health could be assessed. By means of an analysis of variance model for persons nested within families, evidence for familial aggregation of plaque index was found both before and after adjustment for covariates such as age, gender, race, and reported oral hygiene habits. While a substantial fraction of variance in gingival index and attachment loss was also due to differences among families, neither attained statistical significance in these data. Examination of familial correlations (e.g., parent-offspring, sib sib, spouse correlations) confirmed that plaque index showed greater familial resemblance compared with other measures of periodontal health. Both mean gingival index and mean attachment loss showed a stronger correlation between mothers and offspring compared with fathers and offspring. This suggests that further analysis of models for separating genetic and environmental effects may be appropriate for plaque index, but complete analysis of other periodontal indices will require more flexible statistical models for separation of genetic and cultural inheritance while considering gender-specific expression and transmission, as well as incorporation of information from covariates. PMID- 8423254 TI - AADS 70th Annual Session. Chicago, Illinois, March 7-10, 1993. Abstracts. PMID- 8423253 TI - Needed: a research agenda for women's oral health. PMID- 8423255 TI - Auditory demonstrations on compact disk for large N. AB - The popular compact disk entitled Auditory Demonstrations, sponsored by the Acoustical Society of America, includes a number of demonstrations that lead to quantitative results. Those demonstrations are evaluated here in the context of a sizeable class in a lecture room. Demonstrations concern masking, loudness, and pitch; specifically they are numbers 2, 3, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 20, and 25. The evaluations find that most of the demonstrations successfully illustrate psychoacoustical principles in a classroom context; others are less successful or require special circumstances for success. Explanations for success and failure are offered, together with some suggestions for optimizing the chances of success. PMID- 8423256 TI - Two-tone suppression of inner hair cell and basilar membrane responses in the guinea pig. AB - Recordings of receptor potentials from inner hair cells (IHCs) and the basilar membrane (BM) motion were made in pigmented guinea pigs. The acoustic stimuli were single tones near best frequency (BF) and two-tone complexes. Single tone input/output (I/O) functions had a saturating growth for the magnitude and their phase shifts were strongly dependent on the tone frequency relative to BF. For IHCs, a BF tone stimulus produced no phase shift in the ac receptor potential response. Phase lag or lead occurred for tones below or above BF, respectively. BM velocity I/O functions were not as compressively saturating as IHC ac I/O curves. BM phase shifts (in relation to BF) were similar to those of the IHCs. Two-tone suppression was observed in both IHC and BM response measures. Suppressor tones on the low-frequency side of BF produced complex suppression results, which were inconsistent with a simple attenuation model for suppression. The growth of suppression was faster than the attenuation from equivalent level reductions of the probe tone, and phase shifts were phase lead. Depending upon experimental conditions, phase change with suppression may be in the opposite direction from phase change observed from pure attenuation of the probe tone. High-frequency suppressors (relative to BF) are consistent with an attenuation model of suppression for the IHCs of the current study. High side suppression of basilar membrane velocity, however, differed from the IHCs in a systematic way. The phase change caused by suppression of BM velocity was always smaller than that of an equivalent reduction in the probe tone level. PMID- 8423257 TI - A model for the responses of low-frequency auditory-nerve fibers in cat. AB - A computational model was developed for the responses of low-frequency auditory nerve (AN) fibers in cat. The goal was to produce realistic temporal response properties and average discharge rates in response to simple and complex stimuli. Temporal and average-rate properties of AN responses change as a function of sound-pressure level due to nonlinearities in the auditory periphery. The input stage of the AN model is a narrow-band filter that simulates the mechanical tuning of the basilar membrane. The parameters of this filter vary continuously as a function of stimulus level via a feedback mechanism, simulating the compressive nonlinearity associated with the mechanics of the basilar membrane. A memoryless, saturating nonlinearity and two low-pass filters simulate transduction and membrane properties of the inner hair cell (IHC). A diffusion model for the IHC-AN synapse introduces adaptation. Finally, a nonhomogeneous Poisson process, modified by absolute and relative refractoriness, provides the output discharge times. Responses to several different stimuli are presented. These responses illustrate nonlinear temporal response properties that cannot be achieved with linear models for AN fibers. PMID- 8423258 TI - Vibrotactile adaptation enhances amplitude discrimination. AB - Human psychophysical detection and amplitude discrimination thresholds for 25-Hz sinusoidal vibrations were measured on the thenar eminence using two-interval forced-choice tracking, in the unadapted state and following exposure to 25-Hz adapting stimuli representing a range of amplitudes (5-25 dB SL). As expected, detection threshold was elevated 6 to 7 dB for each 10-dB increase in the adapting stimulus. In contrast, amplitude difference thresholds for 10 and 20 dB SL standard stimuli were generally lowest when the amplitude of the adapting stimulus was equal to the amplitude of the standard. The results indicate that while adaptation impairs detection of a liminal vibrotactile stimulus, it improves intensity discrimination of supraliminal stimuli that are close in amplitude to the adapting stimulus. The compatability between these results and a recently proposed model of cortical dynamics (Whitsel et al., 1989) suggests that cortical events may contribute significantly to the physiological basis of vibrotactile adaptation. PMID- 8423259 TI - Intensity just-noticeable differences at equal-loudness levels in normal and pathological ears. AB - The relationship between loudness level and intensity discrimination was investigated in the frequency range 0.5-6.5 kHz by comparing the intensity just noticeable differences (jnd's) at a number of equal-loudness levels in the better and poorer ears of eight individuals with essentially unilateral hearing loss of cochlear origin. Such hearing loss usually produces loudness recruitment, altering the relationships among the loudness, sound-pressure level (SPL), and sensation-level (SL) variables, so that their effects can be separated. In these experiments, the jnd's were correlated with loudness level, not SPL or SL. Although, in some listeners, there were significant differences between jnd's in the two ears at some binaurally equal-loudness levels, they were not systematic over the group of listeners. Statistically, there was no significant difference between equal-loudness jnd's in the two ears, whereas the corresponding SLs and SPLs were significantly different. The rate of loudness growth had no effect on the jnd's. As expected from the near-miss to Weber's law, the mean jnd's decreased with sound intensity. The corresponding decrease in the mean Weber fraction, delta I/I, was accompanied by a decrement in its intrasubject standard deviation. The latter was correlated with the mean, independent of its relationship to SPL or SL. PMID- 8423260 TI - Comodulation masking release in subjects with unilateral and bilateral hearing impairment. AB - Three subjects with unilateral cochlear hearing loss and three subjects with bilateral cochlear hearing loss were tested in three experiments. In the first, their auditory filter shapes were measured for center frequencies of 700 and 2000 Hz, using the notched-noise method. The auditory filters were generally broader for the impaired than for the normal ears. In experiment 2, the threshold for detecting a 2000-Hz signal centered in a band of noise was measured as a function of the noise bandwidth for a Gaussian noise, and for that same noise multiplied (modulated) by a second noise low-pass filtered at 12.5 Hz. For the Gaussian noise, thresholds increased up to a certain noise bandwidth and then flattened off. This bandwidth was usually greater for the impaired than for the normal ears, consistent with the broader auditory filters of the impaired ears. For the modulated noise, thresholds tended to decrease when the noise bandwidth was increased beyond a certain value, indicating comodulation masking release (CMR). The decrease occurred at wider bandwidths for the impaired than for the normal ears. For the unilaterally impaired subjects, the amount of decrease was smaller for the impaired than for the normal ears when tested at equal SPL, but not when tested at equal SL. In experiment 3, the threshold for detecting a 700-Hz signal centered in a 20-Hz-wide band of noise (the on-frequency band, ONB) was measured in the presence of eight flanking bands (FBs) whose envelopes were either identical with that of the ONB (correlated condition) or were uncorrelated. CMR was defined as the difference in threshold between the correlated and uncorrelated conditions. The ONB and the FBs were presented either to the same ear (monaural condition) or to opposite ears (dichotic condition). CMRs tended to be greatest at high levels of the ONB and the FBs. CMRs in the monaural condition were smaller for hearing-impaired than for normal ears. However, at high levels, CMRs in the dichotic condition were similar for normal, bilaterally impaired, and unilaterally impaired subjects. In the latter case, CMRs were similar when the ONB was presented to the normal ear and to the impaired ear of each subject.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423261 TI - Audibility of partials in inharmonic complex tones. AB - These experiments examined the ability of musically trained subjects to hear out individual partials in complex tones with partials uniformly spaced on a scale related to the equivalent rectangular bandwidth (ERB) of the auditory filter. ERB spacings of 0.75, 1.0, 1.25, 1.5, and 2 were used, and the central component always had a frequency of 1000 Hz. All components had a level of 65 dB SPL. On each trial, subjects heard a pure tone (the "probe") followed by a complex tone. The probe was close in frequency to one of the partials in the complex, but was mistuned downward by 4.5% on half the trials (at random) and mistuned upward by 4.5% on the other half. The task of the subject was to indicate whether the probe was higher or lower in frequency than the nearest partial in the complex. The partial that was "probed" varied randomly from trial to trial. Scores for the highest and lowest components in the complexes were generally high (> 90%) for component spacings greater than 1 ERB, but worsened somewhat for ERB spacings of 0.75 and 1.0. Scores for the inner components were close to chance level at 0.75 ERB spacing, and improved progressively as the ERB spacing was increased from 1 to 2 ERBs. For ERB spacings of 1.25 or less, the scores did not change smoothly with component frequency; marked irregularities were observed, as well as systematic errors. An explanation for these is suggested in terms of irregularities in the middle ear transfer function. Performance for the inner components tended to be worse for component frequencies above 1000 Hz than below 1000 Hz. It is suggested that this happens because the pitches of partials are partly coded in the time patterns of neural impulses (phase locking), and the precision of phase locking deteriorates progressively with increasing frequency above 1000 Hz. The auditory filter shapes of the subjects were measured for a center frequency of 1000 Hz, using the notched noise method. One subject had a broader auditory filter than the other three subjects, and this same subject generally had more difficulty in hearing out partials from complex tones than the other subjects. PMID- 8423262 TI - Frequency discrimination in noise: comparison of cat performances with auditory nerve models. AB - Pure-tone frequency discrimination (delta F) performances were measured in cats and compared to neural models of these delta F performances based on auditory nerve data in cats. Animal psychophysical techniques were used to train cats to discriminate frequency changes for pulsed pure tones in background noise at both 1.0 and 3.0 kHz. A go-left, go-right procedure was employed, and delta F's were measured in noise as a function of signal level at a constant signal-to-noise ratio. In contrast to human listeners, cats showed increases in delta F at 1.0 kHz with increasing signal level. Model estimates of delta F's based on rate responses in the cat auditory nerve predict increasing delta F with increasing signal level, the trend observed in the cat psychophysical data. Model estimates of delta F's based on temporal (phase-locking) properties in cat auditory nerve, on the other hand, predict decreases in delta F that have been observed in previous data from human listeners [Dye and Hafter, J. Acoust. Soc. Am. 67, 1746 1753 (1980)]. These results suggest that for cats, average rate, rather than phase-locking, may be used by the central nervous system in performing frequency discrimination in background noise at 1.0 kHz. At 3.0 kHz cats showed little change in delta F as a function of signal level, a result similar to the trend for human listeners to show no change or slight increases in delta F with increases in signal level for tones in the 2- to 3-kHz range. PMID- 8423263 TI - Interaural temporal discrimination using two sinusoidally amplitude-modulated, high-frequency tones: conditions of summation and interference. AB - This paper concerns sensitivity to interaural temporal delays (ITD) in the envelopes of two, sometimes simultaneously presented, sinusoidally amplitude modulated (SAM) tones. The SAM tones were fully modulated (typically at a rate of 250 Hz) and had carrier frequencies of either 2 or 4 kHz. Of particular interest were cases in which the delay to be detected (target ITD) occurred in only one spectral region or in both. The reference ITD, to which the target ITD was added, was either 0, 300, or 600 microseconds. There were three general outcomes: (1) When both regions contained a target ITD, there was an improvement in sensitivity that was quantitatively consistent with an optimal use of independent information. This type of summation was seen even when the target ITDs were added to a pair of SAM tones with two different reference ITDs; (2) when the target ITD was restricted to the higher spectral region, sensitivity was reduced, indicating interference. Interference occurred when the two spectral regions had the same or different reference ITDs and the same or different rates of modulation; (3) sensitivity was unaffected (i.e., no interference occurred) when the target ITD was restricted to the lower spectral region of the pair of SAM tones. These results supplement the observations of Buell and Hafter [J. Acoust. Soc. Am. 90, 1894-1900 (1991)], who used low-frequency tones. PMID- 8423264 TI - Focused attention during selective adaptation along a place of articulation continuum. AB - The current investigation manipulated subjects' attention to adaptor tokens in five selective adaptation experiments. All stimuli were synthetic consonant-vowel syllables, with the consonant varying from [b] to [d] by formant frequency transitions. Two distractor conditions (auditory and visual) were compared to a more typical endpoint-[d alpha] adaptor condition. Distraction from endpoint-[d alpha] adaptors to phonetically distinct [si] and [integral of i] was used to observe whether smaller adaptation effects would result when attention was not focused on adaptor stimuli. In contrast, a focused attention condition required subjects to whisper [b alpha] adaptors right after they were heard. Performance in the focused attention condition was compared to a more typical endpoint-[b alpha] adaptation condition. Results indicated that focused attention did not affect the size of the adaptation effect. Asymmetrical adaptation results for [d alpha] vs [b alpha] adaptors, and a larger amount of adaptation with the presence of contralateral "distractor" syllables, resembled findings in psychoacoustic studies of discrimination and loudness adaptation. These results suggest that two levels of auditory processing (not special to speech perception) were responsible for the observed adaptation effects. PMID- 8423265 TI - A model for context effects in speech recognition. AB - A model is presented that quantifies the effect of context on speech recognition. In this model, a speech stimulus is considered as a concatenation of a number of equivalent elements (e.g., phonemes constituting a word). The model employs probabilities that individual elements are recognized and chances that missed elements are guessed using contextual information. Predictions are given of the probability that the entire stimulus, or part of it, is reproduced correctly. The model can be applied to both speech recognition and visual recognition of printed text. It has been verified with data obtained with syllables of the consonant vowel-consonant (CVC) type presented near the reception threshold in quiet and in noise, with the results of an experiment using orthographic presentation of incomplete CVC syllables and with results of word counts in a CVC lexicon. A remarkable outcome of the analysis is that the cues which occur only in spoken language (e.g., coarticulatory cues) seem to have a much greater influence on recognition performance when the stimuli are presented near the threshold in noise than when they are presented near the absolute threshold. Demonstrations are given of further predictions provided by the model: word recognition as a function of signal-to-noise ratio, closed-set word recognition, recognition of interrupted speech, and sentence recognition. PMID- 8423266 TI - The Lombard reflex and its role on human listeners and automatic speech recognizers. AB - Automatic speech recognition experiments show that, depending on the task performed and how speech variability is modeled, automatic speech recognizers are more or less sensitive to the Lombard reflex. To gain an understanding about the Lombard effect with the prospect of improving performance of automatic speech recognizers, (1) an analysis was made of the acoustic-phonetic changes occurring in Lombard speech, and (2) the influence of the Lombard effect on speech perception was studied. Both acoustic and perceptual analyses suggest that the influence of the Lombard effect on male and female speakers is different. The analyses also bring to light that, even if some tendencies across speakers can be observed consistently, the Lombard reflex is highly variable from speaker to speaker. Based on the results of the acoustic and perceptual studies, some ways of dealing with Lombard speech variability in automatic speech recognition are also discussed. PMID- 8423267 TI - The decrease in severity of asthma in children of parents who smoke since the parents have been exposing them to less cigarette smoke. AB - BACKGROUND: In 1985 we became aware that the smoking of parents aggravates their children's asthma. Since then we have advised all referring doctors to urge parents not to expose their asthmatic children to smoke. METHODS: We investigated 807 nonsmoking asthmatic children, from 1 through 17 years of age, who were consecutively referred between 1983 and 1990. We compared the children who were seen before July 1986 with those seen after that date. RESULTS: Those seen in the later period had intimate exposure to a far smaller number of cigarettes smoked per day, both by mothers (7 vs 3, p = 0.005) and by fathers (5 2, p = 0.001). A concurrent improvement was observed in adjusted measures of asthma severity in their children (asthma score 7.5 vs 6.5, p = 0.047; forced expiratory volume in 1 second as a percent of predicted [FEV1%] 79.2 vs 93.7, p = 0.000; and forced expiratory flow rate during middle half of forced vital capacity [FEF25%-75%] 67.3 vs 82.0, p = 0.009), and for every cigarette less smoked in the room with the child the FEV1 increased by 3%. When parents of those seen in the later period were asked whether they had been told that smoke would aggravate their child's asthma, 80% affirmed that they had. The difference in asthma severity between the two time periods was much less in children of nonsmokers than in children of smokers. CONCLUSION: It appears that if parents are aware that smoke will aggravate their child's asthma, the child will be exposed to fewer cigarettes, and the asthma will be less severe. PMID- 8423269 TI - Methotrexate in asthma--an alternative view. PMID- 8423268 TI - Defective antipneumococcal polysaccharide antibody response in children with recurrent respiratory tract infections. AB - BACKGROUND: Recurrent pyogenic infections are known to occur in patients with an impaired response to polysaccharide antigens. We investigated the occurrence of deficient responses to pneumococcal capsular polysaccharides in patients with recurrent respiratory tract and recurrent systemic infections. METHODS: Forty five patients, 1.7 to 17.1 years of age, were immunized with 23-valent pneumococcal polysaccharide vaccine. Antibody levels to seven pneumococcal serotypes (3, 4, 6A, 9N, 14, 19F, 23F) were determined by ELISA before and after immunization. In addition, patients received a booster immunization with diphtheria toxoid, tetanus toxoid, and poliomyelitis virus vaccine. RESULTS: Thirty-five patients had normal serum immunoglobulin levels. Five of these patients (14%) had low antipneumococcal preimmunization antibody levels and failed to respond to pneumococcal vaccination, whereas the response to booster immunization with protein antigens was appropriate. Three patients were younger than 3 years old, and one had a family history of IgG2 deficiency. Low IgG developed in a fifth patient during follow-up. Ten patients had a humoral immunodeficiency. Seven of these patients failed to respond to pneumococcal vaccination. CONCLUSIONS: We conclude that a defective immune response to polysaccharide antigens in patients requires long-term follow-up to distinguish transient maturational delay from a persistent selective impaired response to polysaccharide antigens, which on occasion may precede the development of humoral immunodeficiency disease. PMID- 8423270 TI - Reorganization of the American Academy of Allergy and Immunology--1987. PMID- 8423271 TI - The Academy reorganizes on its fiftieth anniversary. PMID- 8423272 TI - The American Academy of Allergy: an historical review. PMID- 8423273 TI - Allergens in Hymenoptera venom XXIV: the amino acid sequences of imported fire ant venom allergens Sol i II, Sol i III, and Sol i IV. AB - The most common cause of insect venom allergy in the Southeastern United States is the imported fire ant. The allergens are among the most potent known, with nanogram doses causing sensitization and provoking anaphylaxis. The complete amino acid sequences of imported fire ant venom allergens, Sol i II, III, and IV, were determined by solid-phase protein sequencing of overlapping peptide fragments. Sol i II has a single sequence of 119 amino acids and a molecular weight of 13,217. It has seven cysteine residues, and in its native form is a disulfide-linked dimer. The highly purified molecule does not have phospholipase activity and is not structurally related to phospholipases or other known proteins. Sol i IV has 117 amino acids, for a molecular weight of 13,340. It has six cysteines and is a monomer. Its sequence is 35% identical to Sol i II, but it is not significantly related to other proteins. The Sol i IV sequence showed two amino acid variations. Sol i III was found to consist of 212 amino acids of molecular weight 24,040 in good agreement with 26,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The sequence contained eight cysteine residues and was found to be 44% to 50% identical to five vespid venom antigen 5 molecules. IgE antibodies against Sol i III do not exhibit strong cross reactivity with vespid antigen 5s. The sequence similarity is consistent with other data, suggesting that ants are related to wasps of the superfamily Vespoidea.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423274 TI - The evidence of platelet activation in bronchial asthma. AB - The possible involvement of platelets in bronchial asthma was investigated under three different conditions: (1) chronic asthma, (2) bronchial provocation inhaling house dust mite (HDM), and (3) status asthmaticus. Plasma levels of beta thromboglobulin (beta-TG), platelet factor 4 (PF4), and in part, platelet activating factor (PAF) were measured. Approximately one third of the subjects with symptomatic or asymptomatic chronic asthma showed an increased level of beta TG or PF4. Statistically significant differences occurred in beta-TG and PF4 levels only between healthy controls and symptomatic subjects. Five out of six subjects showed no elevation of beta-TG and PF4 during immediate asthmatic response. In two out of nine subjects with status asthmaticus, beta-TG or PF4 was elevated, and statistically significant correlations occurred between the initial level of PAF and that of beta-TG or PF4. Those results suggest that the platelet activation in the circulation is sometimes provoked in asthma, but plasma level of alpha-granule-derived proteins does not reflect the intensity or severity of asthma, and that PAF is likely to be a mediator responsible for the platelet activation. PMID- 8423275 TI - Treatment of allergic rhinitis with intranasal corticosteroids in patients with mild asthma: effect on lower airway responsiveness. AB - The effect of treatment of allergic rhinitis with intranasal corticosteroids on lower airway responsiveness was assessed in a randomized, double-blind, placebo controlled, crossover study. Twenty-one young patients with perennial allergic rhinitis and asthma, with documented lower airway hyperresponsiveness (PC20 methacholine < 8 mg/ml), were treated with intranasal aqueous beclomethasone dipropionate and placebo, each given for 4 weeks. Patients recorded rhinitis and asthma symptom scores and monitored peak expiratory flow rates every morning and evening. Patients recorded global assessment of rhinitis and global asthma symptom scores at the beginning and end of each treatment. PC20 methacholine was performed at baseline and at the end of each treatment period. Intranasal beclomethasone dipropionate significantly reduced global rhinitis symptom scores (p = 0.05) after 4 weeks of treatment. Global asthma scores did not change significantly (p = 0.2). Geometric mean PC20 methacholine improved significantly after 4 weeks of intranasal beclomethasone, but not after placebo (p = 0.04). Daily morning and evening rhinitis symptom scores were lower in patients treated with intranasal corticosteroids over the first 4 weeks of treatment, but carryover effect of steroids precluded comparative analysis of the second 4-week block (morning p = 0.06, evening p = 0.03). Morning asthma scores tended to decrease (p = 0.07). Evening asthma scores were significantly decreased at weeks 2 and 3 (p = 0.001, p = 0.02, respectively). No change in peak expiratory flow rate was seen. This study confirms that treatment of inflammation in the upper airways indirectly improves asthma symptoms and decreases bronchial hyperreactivity. Ignoring inflammation in the upper airway may lead to suboptimal results in asthma treatment. PMID- 8423276 TI - Final food labeling regulations. PMID- 8423277 TI - Achieving excellence in dietetics practice: certification of specialists and advanced-level practitioners. PMID- 8423278 TI - Using Q-methodology in program evaluation: a case study of student perceptions of actual and ideal dietetics education. AB - Education and program evaluation have rarely included students or clients. Their role in evaluation can be facilitated, however, by Q-methodology. In this article, Q-methodology is described, an illustrative case study is presented, and applications in dietetics are discussed. The case study is a program evaluation of The American Dietetic Association's new Standards of Education (SOE) from the student perspective. The SOE emphasize outcome, whereas the former Essentials of Education (EOE) emphasized process. Students from four programs using the SOE and five programs using the EOE completed two Q-sorts; in one they described their perceptions of their actual program, and in the other their perceptions of an ideal program. Students in SOE programs accurately perceived the outcome emphasis of their actual programs and students in EOE programs perceived the process emphasis of their actual programs. Students in both programs perceived an ideal program as process oriented. In SOE programs, the ideal was so different from the actual that we concluded that students were dissatisfied with their programs. In EOE programs, the views of actual and ideal programs were similar; thus, students seemed satisfied with their programs. The case study illustrates ways students can participate in program evaluation. PMID- 8423279 TI - Retrospective analysis of the use of a standardized reference form in admissions to a dietetics internship program. AB - Although the letter of recommendation is the most commonly requested information relating to the personal qualities of applicants to dietetics internship programs, little research has focused on its value in selection decisions. The purpose of this study was to review how 318 letters of recommendation submitted on a standardized form related to the source of the reference and to the admission status of the applicants. The form contained 40 attributes that raters assigned to one of six categories. Nine of the 40 attributes were not rated by more than 75% of the raters, and 3 of the attributes were rated as outstanding by more than 60% of the raters. We concluded that these attributes did little to distinguish among applicants. The attribute maturity correlated 0.70 with 5 attributes and 0.99 with 2 attributes, so duplication of information existed. Raters were categorized as follows: adviser, major professor, other professor, employer, and other. The highest mean ratings were given by advisers; major professors rated students lowest. Analysis of variance supported a significant difference in rating by type of rater. Our findings suggest that fewer items should be used on a standardized form and that the type of rater should be specified if references are to distinguish among applicants. PMID- 8423280 TI - Prenatal WIC participation can reduce low birth weight and newborn medical costs: a cost-benefit analysis of WIC participation in North Carolina. AB - A number of previous studies have found that prenatal participation in the Special Supplemental Food Program for Women, Infants, and Children (WIC) improves birth outcomes, but only a few studies have provided cost-benefit analyses. The present study linked Medicaid and WIC data files to birth certificates for live births in North Carolina in 1988. Women who received Medicaid benefits and prenatal WIC services had substantially lower rates of low and very low birth weight than did women who received Medicaid but not prenatal WIC. Among white women, the rate of low birth weight was 22% lower for WIC participants and the rate of very low birth weight was 44% lower; among black women, these rates were 31% and 57% lower, respectively, for the WIC participants. Multivariate logistic regression analysis confirmed that prenatal participation in a WIC program reduced the rate of low birth weight. It was estimated that for each $1.00 spent on WIC services, Medicaid savings in costs for newborn medical care were $2.91. A higher level of WIC participation was associated with better birth outcomes and lower costs. These results indicate that prenatal WIC participation can effectively reduce low birth weight and newborn medical care costs among infants born to women in poverty. PMID- 8423281 TI - Vitamin C status in elderly women: a comparison between women living in a nursing home and women living independently. AB - The vitamin C status in blood fractions in 135 elderly women aged 65 years and older was studied within the framework of the Dutch Nutrition Surveillance System. Mean (+/- standard deviation) vitamin C intake (mg/day) was lower among women living in a nursing home (54 +/- 27 mg/day) than among women living in service flats (97 +/- 55 mg/day) and women living independently (132 +/- 44 mg/day). (Service flats are apartments in which the rent includes housekeeping and, when ordered, meal service.) Marginal vitamin C values (< 23 mumol/L) in blood fractions and even levels as low as those found in clinical scurvy (< 11 mumol/L) were frequently observed. In the nursing home, 35% of the women had plasma vitamin C values below 11 mumol/L, and 23% had values between 11 and 23 mumol/L. Blood levels were not significantly affected by age, smoking status, or use of particular drugs but were strongly (r = .47 or, after logarithmic transformation, r = .64) associated with daily intake of vitamin C. Low intake of vitamin C resulted from an overall low food consumption and selective restriction of food products rich in vitamin C. Vitamin C losses caused by food preparation practices and distribution in the nursing home's catering system reduced actual vitamin C intake levels but these losses were not substantially greater than those that are assumed to occur as a result of preparation practices by women living independently. PMID- 8423282 TI - Educational empowerment of the clinical dietitian: a proposed practice doctorate curriculum. AB - Practice doctorate (NutritionD) programs that emphasize use of discipline specific diagnostic assessments have been proposed for clinical dietitians. A preliminary survey of dietitians was conducted to ascertain perceived barriers to use of dietetics-specific diagnostic assessments; interest in earning a practice doctorate, and course topics of perceived importance to selected subspecialties and a practice doctorate curriculum. A questionnaire was developed and administered to a nonrandom sample of 120 dietitians attending a clinical nutrition conference. Response rate was 83%, yielding 100 usable responses. Of the respondents, 60 perceived education to be one of the most limiting barriers to use of diagnostic assessments; 55 expressed interest in earning a practice doctorate. Respondents ranked 22 course topics for perceived importance in each of 14 dietetics subspecialties; 5 topics were categorized as core, 12 as supporting, and 5 as elective. Course topics perceived to be of highest importance in 11 or more of 14 subspecialties were nutritional diagnosis; clinical nutrition examination procedures; advanced diet therapies/nutriotherapeutics; drug-nutrient interactions; and the care process/diagnostic charting. Dietetics-specific diagnosis, which often complements medical diagnosis, is in the process of becoming a standard of practice for many dietitians. Therefore, as seasoned dietitians we are challenged to overcome any possible educational barriers to realization of this new practice. One approach to achieving this educational empowerment would be to direct national study efforts toward components of the proposed curriculum and to develop a practice doctorate option for clinical dietitians. PMID- 8423283 TI - Recombinant bovine and porcine somatotropin: safety and benefits of these biotechnologies. AB - This article reviews the literature about the safety and benefits of two recombinantly derived proteins, bovine somatotropin (bST) and porcine somatotropin (pST), that likely will be used in animal agriculture in the future. When administered to dairy cows, bST increases milk production per cow approximately 15% to 20% and improves productive efficiency approximately 10%. Administration of pST to growing pigs reduces carcass fat content by as much as 70% to 80% and improves productive efficiency 15% to 35%. Because meat is a major source of total fat and saturated fatty acids in the diets of human beings, pST will allow consumers to include leaner, more nutrient-dense pork in their diets and still meet current dietary guidelines. Although these biotechnologies have not yet received regulatory approval from the Food and Drug Administration (FDA) for commercial use, information published by the FDA, the National Institutes of Health, the US Congress Office of Technology Assessment, and the American Academy of Pediatrics, as well as an extensive body of scientific evidence, indicate that these products are safe for the consumer. Nonetheless, it is important that consumers understand the benefits and safety of these biotechnologies. Dietitians can play an important role in providing information to consumers about the safety and benefits of bST and pST. PMID- 8423284 TI - The Ohio NSPS statewide survey of third-party reimbursement policies for nutrition services. PMID- 8423285 TI - Personality-focused learning environments: learning preferences of students in dietetics and restaurant, hotel/institution management programs. PMID- 8423286 TI - Job satisfaction of clinical, community, and long-term-care dietitians in New York City. PMID- 8423287 TI - Position of the American Dietetic Association: biotechnology and the future of food. PMID- 8423288 TI - Characteristics of advanced-level dietetics practice: a model and empirical results. AB - This article, which is the first of a two-part series, presents results for the first objective of The American Dietetic Association (ADA) 1991 Dietetic Practice Study: to determine the characteristics of advanced-level dietetics practice. A nationwide mail survey of ADA members was conducted on a stratified random sample of 8,012 beyond-entry-level (registered before April 1988) registered dietitians who were members of dietetic practice groups (DPGs). The sample was supplemented with two randomly selected control groups of 1,000 entry-level and 1,000 beyond entry-level registered dietitians. The overall response rate was 63.1%. The 5,852 usable returns were representative of the dietetics population surveyed. A model of advanced-level professional practice was developed that specified minimum necessary requirements for advanced practitioners on five components: education and experience, professional achievement, approach to practice, professional role positions, and professional role contacts; measurement of a sixth component, advanced-level practice performance, was unsuccessful. A series of validation analyses found the model to be a statistically sound and reliable means of distinguishing advanced practitioners from other groups of dietitians in 8 of every 10 cases. A total of 461 (8.9%) dietitians met all requirements of the model and were classified as advanced practitioners. Projected estimations of advanced practitioners in the population of beyond-entry-level ADA members who are also members of DPGs ranged between 2,126 and 2,640 dietitians (3.5% to 4.3% of the ADA membership).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423289 TI - Specialty practice in dietetics: empirical models and results. AB - This article, which is the second of a two-part series, presents results for the second and third objectives of The American Dietetic Association (ADA) 1991 Dietetic Practice Study: to distinguish the task activities among registered dietitians practicing in renal nutrition, pediatric nutrition, and metabolic nutrition care; and to investigate the relationship between advanced-level practice and specialty practice. A nationwide mail survey of advanced-level practice was conducted on a stratified random sample of 8,012 beyond-entry-level (registered before April 1988) registered dietitians who were members of ADA and of dietetic practice groups (DPGs). The sample was supplemented with two randomly selected control groups of 1,000 entry-level and 1,000 beyond-entry-level registered dietitians. The operational definition for specialty practice was met by 1,925 sample members who were also included in the survey on practice in the three specialty areas. The overall response rate was 63.1%, and the total number of usable returns was 5,852. The results from a discriminant analysis of 121 specialty job tasks, administered in common to practitioners in each specialty, found that 84 tasks could be used to construct a generic model of specialty practice from which specific task lists were derived that defined practice in renal nutrition, pediatric nutrition, and metabolic nutrition care. Validation analyses found the generic model to be a reliable means of distinguishing the job activities in renal nutrition and in pediatric nutrition; it was somewhat less reliable for metabolic nutrition care. A weak relationship was found between advanced-level practice and practice in the three specialty areas.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423290 TI - Report on the 1991 membership database of the American Dietetic Association. PMID- 8423291 TI - Nonunion of the scaphoid: a critical analysis of recent natural history studies. AB - To determine whether treatment of a disorder is effective, a surgeon must first understand the natural history of the disorder without treatment. We used guidelines for the performance of a valid natural history study that appear in the literature and applied them to articles on the natural history of scaphoid nonunion. No study satisfied all criteria. Moreover, none developed an inception cohort, which is the single most important method of a valid natural history study. In each study, the direction of the bias was identifiable: the natural history of scaphoid nonunion was distorted to make the outcome more severe than if methodologic problems had not been present. We conclude that the natural history of nonunion of the scaphoid is not as severe as has been reported in the literature. The magnitude of the difference is not known at this time. PMID- 8423292 TI - Extended survival and function of peripheral nerve allografts after cessation of long-term cyclosporin administration in rats. AB - To determine whether survival and function of peripheral nerve allografts are possible after cessation of long-term cyclosporin (CsA) treatment, we grafted 4.25 cm Lewis rat peripheral nerve allografts (n = 22) into tibial nerve gaps in recipient brown Norway rats. Allograft groups received CsA (15 mg/kg) subcutaneously every day for 20 days and then biweekly for either 5 or 8 months after transplantation. The control group, brown Norway rats with isografts from brown Norway donor rats (n = 2), also received identical CsA treatment. Semimonthly electrophysiologic studies were done from postoperative week 13 until the animals were killed (up to 79 weeks). The corresponding CsA levels in the nerve and blood were recorded from cessation of CsA up to 58 weeks after surgery. No electrophysiologic signs of rejection were observed in any of the 22 allograft recipients treated with CsA for up to 8 months or in 17 of the 22 observed for up to 58 weeks after cessation of CsA. Overall, 5 of 22 allografts were rejected in the first 8 weeks after discontinuation of CsA. Signs of rejection occurred only in the 5-month treatment group and followed the large initial drop in CsA blood level (from 1010 ng/ml to < 25 ng/ml) that occurred within the first 6 weeks after CsA cessation. The two isograft controls demonstrated no electrophysiologic signs of rejection up to 58 weeks after surgery. Peripheral nerve allografts in the rat can regenerate and function on long-term CsA; after cessation of CsA, they can function for extended periods of time without signs of rejection if trace amounts of CsA are present.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423293 TI - A study of the posterior interosseous nerve (PIN) and the radial tunnel in 30 Thai cadavers. AB - Sixty fresh cadaver upper extremities were dissected for a study of the course of the posterior interosseous nerve (PIN) in the radial tunnel, an area that varies but begins in the furrow between the brachioradialis and brachialis in the distal arm and ends at the distal edge of the supinator muscle in the proximal forearm. The radial nerve pierces the lateral intermuscular septum 13 cm above the elbow joint line. At 1.3 cm above the joint line the nerve divides into its posterior interosseous and superficial radial branches. The extensor carpi radialis brevis muscle received nerve supply from the superficial radial nerve, radial nerve, and PIN in 43%, 55%, and 2%, respectively. The PIN passed through the radial tunnel anterior to the radiohumeral joint and then coursed laterally and posteriorly beneath the arcade of Frohse, which is the proximal edge of the superficial layer of the supinator muscle. The arcade was tendinous in 57% of the cadavers and membranous in 43%. The distal edge of the supinator was tendinous in 65% of the specimens and membranous in 35%. The purpose of this article was to measure the various lengths of the PIN and to study possible compression sites at normal variations along the course of the PIN in the radial tunnel. No cases showed evidence of PIN compression in the radial tunnel. PMID- 8423294 TI - Magnetic resonance imaging in the evaluation of persistent carpal tunnel syndrome. AB - Magnetic resonance imaging was used to assess nine median nerves in the wrists of seven patients who had signs and symptoms of persistent compressive median neuropathy despite previous carpal tunnel release. Intraoperative findings were then correlated in eight surgically treated cases with both MRI findings and postoperative results. Magnetic resonance imaging suggested a potential abnormality in each of eight operative cases. These findings correlated very well with both intraoperative observations and postoperative results, which indicated that some abnormality involving either the median nerve or the transverse carpal ligament had been present in all cases. Magnetic resonance imaging proved to be a sensitive and specific tool in the evaluation of persistent postoperative median nerve compression. PMID- 8423295 TI - Radial nerve palsy caused by open humeral shaft fractures. AB - Although radial nerve palsy associated with a closed humeral shaft fracture may be managed by observation, it is our experience that an open humeral shaft fracture with radial nerve palsy requires exploration of the nerve. In a series of 14 patients with radial nerve palsy caused by an open humeral shaft fracture, 9 (64%) of the 14 patients had a radial nerve that was either lacerated or interposed between the fracture fragments. There was an equal incidence of radial nerve lacerations or entrapments in types I, II, and III open humeral shaft fractures. Epineural radial nerve repair, done primarily or secondarily, provided a satisfactory return of radial nerve function. Rigid fixation of the associated fracture is the recommended treatment. PMID- 8423296 TI - High-pressure injection injuries of the hand: review of 25 patients managed by open wound technique. AB - This article reviews the outcome of injury of the hand in 25 patients treated at our institution by the open wound technique: wide debridement, drainage, open packing, and delayed closure. The results obtained are superior to those reported in the literature: Among the patients, 84% of the involved hands or fingers were salvaged, 64% had essentially normal hand function at the time of final follow up, and 92% were able to return to their previous jobs. We propose that aggressive treatment by the open wound technique improves the outcome of high pressure injection injuries of the hand. PMID- 8423297 TI - Subcutaneous infection of the hand due to Mycobacterium nonchromogenicum. PMID- 8423298 TI - Ulceroglandular and pulmonary tularemia: a case resulting from a cat bite to the hand. PMID- 8423299 TI - Osteoblastoma of the hamate bone: a case report. PMID- 8423300 TI - Load mechanics of the midcarpal joint. AB - The midcarpal joint was studied in upper extremities of fresh cadavers with the use of a static positioning frame, pressure-sensitive film, and a microcomputer to analyze the contact prints. The contact area on the proximal side of the midcarpal joint was found to consist generally of four areas; the scaphoid trapezium-trapezoid, the scaphoid-capitate, the lunate-capitate, and the triquetrum-hamate. The contact areas accounted for less than 40% of the available joint surface, even under loads of 118 pounds. The distribution of load through the midcarpal joint was scaphoid-trapezium-trapezoid 23%, scaphoid-capitate 28%, lunate-capitate 29%, and triquetrum-hamate 20%. The midcarpal joint, like the radiocarpal joint, appears to transmit load through distinct areas and through a relatively small portion of the available joint surface. PMID- 8423301 TI - Hand manifestations of oxalosis. AB - Oxalosis is an unusual metabolic disease that results either from an inherited hepatic enzyme deficiency or as the result of poor oxalate clearance during chronic hemodialysis. We present two cases of oxalosis and describe the hand manifestations of this condition and their treatment. One patient had painful, progressive gangrene, whereas in the other the disease took an indolent course with small palmar crystalline deposits. PMID- 8423302 TI - Measurement of hand microvascular blood flow with isolated cold stress testing and laser Doppler fluxmetry. AB - Isolated cold stress testing applies cold thermal stress to an extremity for noninvasive study of skin surface temperature as an index of digital microcirculation. In this study, laser Doppler fluxmetry was combined with isolated cold stress testing to examine the relationship between digital thermoregulation (measured by isolated cold stress testing) and digital microcirculatory blood flow (measured by laser Doppler fluxmetry) in 50 healthy men and women. After a 5-minute baseline period, both hands were subjected to cold stress (average, 8 degrees C) for 20 minutes and then returned to room temperature for 20 minutes. Digital temperatures and laser Doppler measurements were graphed to document individual responses. The 25 women had lower digital temperatures, lower laser Doppler fluxmetry values, and less average rewarming than the 25 men. Statistical cluster analysis revealed a cold pattern, found predominantly in women, and a warm pattern, found predominantly in men. These patterns of response may help to elucidate the pathogenesis of the vascular occlusive syndromes occurring predominantly in women. PMID- 8423303 TI - Congenital pseudoaneurysm of the superficial palmar arch in a child: a case report. AB - Both true and false aneurysms are rare in the upper extremity and even more so in pediatric patients. When the condition is congenital, except for those aneurysms found in the circle of Willis, existence in the upper extremity is almost unknown. The following case report describes a congenital pseudoaneurysm involving the superficial palmar arch in a child. PMID- 8423304 TI - Extremity transplantation: a review of its current state of development. AB - The discovery of a powerful immunosuppressive, cyclosporin A, together with the development of a microsurgical free tissue transfer technique, has brought extremity transplantation closer to being a realistic possibility. In this review the authors describe the history of laboratory research into extremity transplantation before immunosuppression, with early immunosuppressive agents, with cyclosporin A, and with FK-506, one of a new generation of immunosuppressives. PMID- 8423305 TI - Dynamic external finger fixator for fracture dislocation of the proximal interphalangeal joint. AB - The treatment of fracture dislocations of the proximal interphalangeal joint often results in pain and stiffness. A small dynamic external finger fixator was designed to maintain the reduced position of the dislocated middle phalanx and allow early active range-of-motion exercise. Four patients with acute unstable fracture dislocations and three with old malunited fracture dislocations of the proximal interphalangeal joint were treated with this apparatus. The average range of the proximal interphalangeal joint motion with this device was 88 degrees. The average follow-up period was 21 months. PMID- 8423306 TI - Temporary orthosis for the cast-immobilized thumb. PMID- 8423307 TI - Silicone arthroplasty of the wrist in rheumatoid arthritis. PMID- 8423308 TI - Detection of foreign bodies in hand. PMID- 8423309 TI - Reflex sympathetic dystrophy and cigarette smoking. PMID- 8423310 TI - Possible complication of belt loop pulley reconstruction. PMID- 8423311 TI - Accuracy of dynamometers. PMID- 8423312 TI - Proximal row carpectomy: a multicenter study. AB - Twenty patients underwent proximal row carpectomy and were retrospectively evaluated for pain, motion, grip strength, functional activity, and x-ray changes at a mean follow-up of 3 1/2 years. For nonrheumatoid patients, motion decreased 15% after surgery, mean grip strength improved 22%, and 82% believed their conditions were improved and said they would repeat the procedure. The procedure failed in all three patients with rheumatoid arthritis. Patients with mild preoperative arthritic changes had better results than those with advanced disease. PMID- 8423313 TI - Scaphocapitolunate arthrodesis. AB - Since 1985 scaphocapitolunate arthrodesis has been performed on 21 patients with either chronic incompetence of the scapholunate ligament or a scaphoid nonunion. The average age was 31 years; follow-up averaged 28 months. Eighty-six percent of the injuries involved workers' compensation. The operative procedure was done through a dorsal approach, with the use of autogenous bone grafting and Kirschner wire fixation. Eight-one percent healed after the primary procedure; one patient had a major infection. Range of motion averaged 35 degrees of extension, 30 degrees of flexion, 10 degrees of radial deviation, and 20 degrees of ulnar deviation. Grip strength averaged 70% of the uninvolved side. Pain was significantly reduced in 80% of the patients. Sixteen of 20 patients were able to return to work. Radiographic examination at final follow-up showed mild degenerative changes at the radiocarpal joint in two patients. PMID- 8423314 TI - Severe fractures of the distal radius: effect of amount and duration of external fixator distraction on outcome. AB - Although severe intra-articular fractures of the distal radius are often treated with external fixation/distraction, little attention has been devoted to the amount and duration of fixator distraction required to maximize outcome. To determine these effects, we conducted a retrospective study of 26 patients whose primary treatment was external fixation/distraction. Patients were evaluated by chart review, questionnaire, x-ray films, and physical examination an average of 104 weeks after injury. The carpal height index was used to quantify distraction. Scores for pain, function, wrist motion, and grip strength and the total score were adversely affected in proportion to the increase in carpal height index by distraction. Outcome was adversely affected as the duration of distraction increased. Motion scores were affected most. Overall, patients recovered at least 75% of motion and grip strength, and 85% attained New York Orthopedic Hospital grades of good or excellent. These are the first data to show that there are potential adverse effects from increasing amounts of distraction and prolonged use of the external fixator. PMID- 8423315 TI - Diagnosis of occult scaphoid fractures by intrasound vibration. AB - We investigated a new means of diagnosing occult scaphoid fractures. Eighty-six patients underwent vibratory testing at presentation, while the clinical examination and standard four-view x-ray examination findings were unknown to the persons who performed the vibratory testing of both the injured and uninjured wrists. Thirty-six patients had radiographically confirmed scaphoid fractures and, after their vibratory tests, were eliminated from the study. Fifty patients, 39 men and 11 women, were believed to have scaphoid fractures on the basis of history and clinical examination findings but were included in the occult scaphoid study group because standard four-view x-ray films of the wrists did not reveal a scaphoid fracture. Distinction between the fracture and no-fracture patients was made with a limited two-phase technetium bone scan and delayed x-ray examination. All patients with known scaphoid fractures (36) had positive findings on vibratory examination. Vibratory testing identified all six of the patients with occult scaphoid fractures (sensitivity 100%). Results of two examinations were false-positive, and none were false-negative (specificity 95%). One of the patients with false-positive results had a fracture of the trapezium, and the other had reflex sympathetic dystrophy. The vibratory testing of injured wrists is inexpensive, noninvasive, and easy to perform, and it involves no ionizing radiation. PMID- 8423316 TI - Cineradiographic study of wrist motion after fracture of the distal radius. AB - Residual dorsal tilt is one of the main deformative factors in the malunion of fractures of the distal radius. We investigated the effect of such tilt on the range of wrist motion and on carpal alignment in a cineradiographic study of wrist motion in 30 patients with extra-articular Colles' fracture. With increased dorsal tilt, the range of wrist motion became more restricted and abnormalities of carpal alignment during wrist motion became apparent. When the dorsal tilt was less than 10 degrees, the effects on the range of motion and on the dynamic carpal alignment were small. PMID- 8423317 TI - The ulnar impaction syndrome: follow-up of ulnar shortening osteotomy. AB - Thirty wrists of 27 patients with ulnar impaction syndrome who underwent ulnar shortening osteotomy were retrospectively reviewed. The average follow-up was 51 months. The wrists were graded preoperatively and postoperatively according to a wrist-grading system modified from Gartland and Werley. The parameters of grading included pain, function, range of motion, grip strength, radiographic analysis, bony union, and complications. Twenty-four wrists were graded excellent, 4 good, 1 fair, and 1 poor after the surgery in comparison with 28 poor and 2 fair before the operative treatment. Complications were rare, with no ulnar nonunions. This long-term follow-up study revealed that distal ulnar shortening osteotomy is an excellent procedure for the treatment of ulnar impaction syndrome. PMID- 8423318 TI - Pronator quadratus transfer for chronic anterior subluxation of the distal ulna: a case report. PMID- 8423319 TI - Recessional ulnar osteotomy: use of reduction forceps for intraoperative radiography and plate application. PMID- 8423320 TI - Two new methods of tendon repair: an in vitro evaluation of tensile strength and gap formation. AB - A new tendon repair design incorporating a mesh sleeve was, together with a new epitendinal suture technique (cross-stitch), tested biomechanically on sheep tendons. The mean breaking strength with a 6-0 suture was 103 N for the mesh sleeve repair and 63 N for the cross-stitch alone, both significantly greater than the 48 N recorded for the modified Kessler repair performed with a 4-0 polyester suture and reinforced with a circumferential conventional 6-0 epitendinal stitch. The cross-stitch grasp was 117% stronger than the conventional epitendinal grasp reinforced by a core stitch. The new techniques effectively prevented the large gaps that occurred in the modified Kessler repair. PMID- 8423321 TI - Rupture of the flexor digitorum profundus tendon in the palm caused by repeated, chronic direct trauma. AB - Nonrheumatoid ruptures of the flexor tendons in the palm are rare. We report a case in which excessive exertion caused rupture of the flexor digitorum profundus tendon to the small finger in the palm at the level of the lumbrical origin. The patient had been practicing Japanese fencing for many years. The hard butt end of a bamboo sword impinged precisely on the region in the palm where the rupture occurred. PMID- 8423322 TI - Fibrin sealant in prevention of flexor tendon adhesions: an experimental study in the rabbit. AB - The effect of fibrin sealant on the development of adhesions in flexor tendon surgery was studied in both immobilized and active weight-bearing paws of rabbits. A partial laceration was produced and was treated with sutures alone or with both sutures and glue. Immobilized tendons treated in these ways showed no difference in adhesion formation, but in mobile tendons there was a difference at 6 weeks in gross appearance, scanning electron micrographic appearance, and strength of adhesions as measured when the distally cut tendon was pulled out of its sheath. These observations indicate that, after repair, a mobile flexor tendon heals with a smooth gliding surface and without formation of restricting adhesions when treated with fibrin glue around the suture site. PMID- 8423323 TI - Morphologic and biomechanical comparison of tendons used as free grafts. AB - The palmaris longus (n = 10), plantaris (n = 11), and extensor digitorum longus (n = 10) tendons were harvested from cadaver limbs for morphologic and biomechanical comparisons. Eleven flexor digitorum profundus tendons from the index finger were also harvested for comparison of biomechanical properties of the free graft tendons with those of a typical digital flexor. Maximal tendon length, cross-sectional area, volume, stiffness, and modulus of elasticity were determined by measurement, laser micrometry, and tensile testing. The plantaris and extensor digitorum longus yielded the longest grafts, 334 and 325 mm, respectively, compared with the palmaris longus, which yielded only 161 mm of length. Cross-sectional areas were as follows: palmaris longus, 3.1 mm2; plantaris, 1.4 mm2; extensor digitorum longus, 3.3 mm2; and flexor digitorum profundus, 10.6 mm2. Mean volumes were as follows: palmaris longus, 529 mm3; plantaris, 557 mm3; and extensor digitorum longus, 1006 mm3. The palmaris longus and the extensor digitorum longus showed greater stiffness, 42.0 and 47.8 N/mm, respectively, than the plantaris, which had a stiffness of 25.5 N/mm. However, the flexor digitorum profundus showed significantly greater stiffness than all of the graft tendons. The modulus of elasticity ranged from 1161.6 to 1673.0 MPa, with no significant difference between tendons tested. The findings in this study provide data that may be useful in the selection of a specific donor for free tendon grafting. PMID- 8423324 TI - Ipsilateral fractures of the scaphoid and radius. AB - From 1985 to 1990, six patients were treated for ipsilateral fractures of the scaphoid and radius. All these injuries resulted from falls from a substantial height, with multiple injuries occurring in all patients. Two of the patients had minimally displaced fractures (1.0 mm), whereas the other four patients had moderately displaced scaphoid fractures (2.0 mm). The treatment for fracture of the distal radius was dependent on the fracture type, but all the scaphoid fractures were treated with internal fixation. All the fractures united, with the radial fractures healing in 6 weeks and the scaphoid fractures healing in an average of 13 weeks. The follow-up time averaged 2.9 years (range, 1.0 to 4.6 years). Internal fixation of the scaphoid in these combined injuries allowed for earlier and more aggressive therapy to maximize wrist and forearm motion. PMID- 8423325 TI - Excursion of prime wrist tendons. AB - To allow continuous and simultaneous measurements of tendon excursion and joint rotation angles, we developed a system using an electric potentiometer and an electromagnetic tracking device. Instantaneous moment arms of each tendon could then be calculated on the basis of the slope of the curve between tendon excursion and joint angular displacement. Five prime wrist motor tendons were studied on freshly frozen cadavers. Calculated tendon moment arms were found to be consistent throughout a full range of flexion-extension wrist motion and radioulnar deviation and corresponded closely to the anatomic location and orientation of the tendons. The flexor carpi ulnaris tendon provided the largest moment contribution to the wrist joint. Forearm rotation did not affect the function of the wrist tendon except for the extensor carpi ulnaris. During flexion-extension motion, the excursion of the extensor carpi ulnaris decreased significantly from 10 to 4 mm, with the forearm in neutral and pronated positions. During radioulnar deviation, the excursion of the extensor carpi ulnaris increased from 14 to 17 mm with the forearm in supinated and pronated positions. PMID- 8423326 TI - Anatomy of the flexor retinaculum. AB - For an accurate definition of the anatomic limits of the carpal tunnel, 26 cadaver upper extremities were studied by gross (10), histologic (3), and radiographic (13) methods. The mean proximal limit of the central portion of the flexor retinaculum was 11 mm distal to the capitate-lunate joint, and the mean distal limit of the distal portion was 10 mm distal to the carpometacarpal joint of the third metacarpal. Carpal tunnel width at the hook of the hamate (20 mm) was significantly smaller than its proximal (24 mm) or distal (25 mm) extent. The flexor retinaculum extended from the distal aspect of the radius to the distal aspect of the base of the third metacarpal. We redefined the palmar boundary of the carpal tunnel to include three continuous segments of flexor retinaculum: the thin proximal segment composed of thickened deep investing fascia of the forearm; the transverse carpal ligament; and the distal portion of the flexor retinaculum, composed of an aponeurosis between the thenar and hypothenar muscles. In light of recent operative procedures that divide only the transverse carpal ligament, this study provides an anatomic basis for a more extensive release. PMID- 8423327 TI - Identification of a novel granulocyte chemotactic protein (GCP-2) from human tumor cells. In vitro and in vivo comparison with natural forms of GRO, IP-10, and IL-8. AB - Stimulated human osteosarcoma cells (MG-63) were used as a source of granulocyte chemotactic protein (GCP). In addition to the previously isolated GCP-1/IL-8, natural forms of GRO alpha, GRO gamma, and IP-10 were purified and identified by amino acid sequence analysis. Further, a novel GCP, GCP-2, was isolated in its natural form (6 kDa) and was found to be structurally related to the other members of the IL-8 family. GRO alpha, IP-10, and GCP-2 showed heterogeneity, in that several forms of each protein were recovered. These differed in truncation at the amino terminus. Reverse phase HPLC allowed us to separate four such different forms of GCP-2. These tumor-derived factors were compared in granulocyte activation and chemotaxis assays. IL-8 induced neutrophil gelatinase B release at 2 nM, but GRO alpha and GCP-2 showed a 5- to 10-fold lower specific activity. When the migration of granulocytes through polycarbonate micropore membranes was measured, GCP-2 and GRO alpha had a maximal chemotactic index comparable to that of IL-8. The minimal effective dose for GCP-2 and GRO alpha was 3 to 10 nM, whereas the specific activity of IL-8 was at least 10-fold higher. IP-10 was not active in this assay at doses up to 100 nM. Finally, in vivo chemotaxis was measured by using granulocyte recruitment in the rabbit skin model. After intradermal injection of 200 ng/site, GCP-2 provoked a significant granulocyte infiltration, albeit to a lesser extent than did IL-8 and GRO alpha. GCP-2 did not attract monocytes in vivo nor did it induce the cells in vitro to migrate or to produce enzyme. In conclusion, this study reveals a new member of the IL-8 family and shows that these related inflammatory mediators possess different potencies and efficacies towards granulocytes. PMID- 8423328 TI - Pertussis toxin-sensitive factor differentially regulates lipopolysaccharide induced tumor necrosis factor-alpha and nitric oxide production in mouse peritoneal macrophages. AB - It has been established that LPS, the major constituent of the outer membrane of gram negative bacteria, stimulates macrophages to produce numerous inflammatory mediators, including TNF-alpha and nitric oxide (NO). Both TNF-alpha and NO are important in the macrophage-mediated cytotoxicity against invading microorganisms and tumor cells. Although many LPS-dependent immune responses have been well characterized phenomenologically, the precise signal transduction pathways in LPS induced macrophage activation are not clear. We reported that 1) pretreatment of C3HeB/FeJ mouse peritoneal macrophages with pertussis toxin (PT) markedly enhanced LPS-induced TNF-alpha production but inhibited LPS-dependent NO production under the same conditions; 2) kinetics of the PT effects on these LPS responses were correlated with PT-mediated ADP-ribosylation of a 41-kDa protein(s); and 3) PT pretreatment did not correct the refractory states of C3H/HeJ macrophages to wild type smooth-LPS. These results suggest that LPS stimulates TNF-alpha and NO production in mouse peritoneal macrophages through different biochemical pathways, and that the signal transduction for both pathways is regulated by a PT-sensitive factor. It is possible that this factor is a guanine nucleotide-binding regulatory protein(s). Finally our data indicate that it is unlikely that the defect of the C3H/HeJ macrophages in response to LPS is at the level of this PT-sensitive factor. PMID- 8423329 TI - A monoclonal antibody to the NH2-terminal segment of human IFN-gamma selectively interferes with the antiproliferative activity of the lymphokine. AB - To gain more information about the relationship between the structure of IFN gamma and its activity, a peptide corresponding to a hydrophilic peak between amino acids 4 and 16 was used to immunize mice and generate mAb. mAb IGMB-15 reacts to both native and rIFN-gamma and neutralizes the antiproliferative activity of IFN-gamma without affecting its antiviral activity or its ability to up-regulate HLA-DR Ag expression. Moreover, we observed that mAb IGMB-15 was unable to inhibit the binding of radiolabeled IFN-gamma to its cellular receptor. These findings show that the NH2-terminal region may somehow be involved in the biologic activity of IFN-gamma. Besides, the capability of mAb IGMB-15 to inhibit the antiproliferative but not the antiviral activity of IFN-gamma in the same cell (HEp-2) suggests the presence of different elements involved in signal transduction, which may account for the multiple activities of the lymphokine. PMID- 8423330 TI - T lymphocytes capable of activating endothelial cells in vitro are present in rats with autoimmune diabetes. AB - Endothelial activation as evidenced by increased expression of leukocyte adhesion molecules occurs during immune-mediated inflammatory processes. One such process is insulitis, the pancreatic islet inflammation that leads to autoimmune insulin dependent diabetes mellitus (IDDM). To determine if the induction of IDDM correlates with the presence of T lymphocytes capable of activating endothelial cells (EC), we studied the diabetes resistant BB (DR) rat. These animals become diabetic after in vivo depletion of T cells expressing the RT6 alloantigen. Various populations of purified DR T lymphocytes were cocultured with MHC compatible rat EC. We observed: 1) RT6- T cells from diabetic animals induced maximal endothelial MHC Ag expression. 2) The ability of RT6- T cells to activate EC increased with the duration of in vivo RT6 depletion. It was acquired before the onset of insulitis but subsided after the onset of diabetes. 3) In contrast, neither unsorted total T cells nor in vitro-purified RT6- T cells activated EC. 4) Older DR rats depleted of RT6+ T cells did not become diabetic and their RT6- T cells did not activate EC. 5) T cell IFN-gamma production correlated with the intensity of EC activation. 6) direct T cell-EC contact was required for maximal IFN-gamma production and EC activation. We conclude that RT6- T cells capable of activating EC are generated during the induction of IDDM in DR rats. We hypothesize that such T cell activity may lead to endothelial activation in vivo and contribute to immune-mediated insulitis, beta-cell destruction, and IDDM. PMID- 8423331 TI - Soluble complement receptor type 1 prevents human complement-mediated damage of the rabbit isolated heart. AB - The purpose of this study was to determine if recombinant human soluble CR1 (sCR1) could prevent tissue damage associated with the activation of human complement. Directly mediated human complement-dependent myocardial injury was induced in the rabbit isolated heart perfused with a Krebs-Henseleit buffer containing 6% human plasma. There were three study groups: 1) 6% heat-inactivated human plasma (control); 2) 6% normal human plasma (NHP); or 3) 6% normal human plasma + 20 nM sCR1 (NHP + sCR1). Recorded functional parameters of the control group remained stable throughout the duration of the 70-min protocol. Complement activation in hearts perfused with 6% NHP increased the diastolic pressure; decreased developed pressure; and increased coronary perfusion pressure. These alterations were accompanied by a decrease in the maximum positive and negative dP/dt. Complement activation also increased cardiac muscle lymphatic fluid flow rate. The changes were greatest between 20 and 40 min, but persisted for the duration of the protocol. sCR1 (20 nM) in the perfusate containing 6% NHP prevented the complement-mediated alterations in the systolic, developed, and coronary perfusion pressures. sCR1 prevented the decrement in the positive and negative dP/dt, and the increase in the lymphatic fluid flow rate. Values for each of these parameters in hearts perfused with 6% NHP + sCR1 were not altered from those of controls at any time point in the protocol. Ultrastructural changes were present in tissues perfused with 6% NHP along with immunohistochemical evidence for presence of the terminal C5b-9 complex. sCR1 prevented the ultrastructural changes and the formation of the terminal complex. sCR1 offers significant protection against the cytolytic effects resulting from activation of the human complement system. PMID- 8423332 TI - Binding of human immunodeficiency virus type-1 to follicular dendritic cells in vitro is complement dependent. AB - The authors studied the binding in vitro of HIV-1 virus particles, conjugated to fluorescein isothiocyanate, to follicular dendritic cells (FDC) isolated from human tonsils. Analysis was done using flow cytometry, fluorescence microscopy, and immunogold electron microscopy. The focus of study was on the effect of serum from various origins, including pooled fresh serum and heated serum from control donors and pooled heated serum from HIV-1-infected patients (containing anti-HIV 1 antibodies). In the presence of heated serum, either from controls or from HIV 1-infected patients, the fluorescence signal in flow cytometry was similar to the background value. In the presence of fresh serum, the signal was substantially increased, and an even higher signal was observed in the presence of fresh serum and serum from HIV-1-infected patients. This high fluorescence signal was also found in the presence of serum depleted of complement factor C5, but not with serum deficient in complement factor C3. The binding of HIV-1 virions to FDC in the presence of fresh serum was confirmed by fluorescence microscopy on cytospot preparations. After quenching of the extracellular fluorescence with trypan blue, the fluorescence was reduced to about 30% of the initial value, indicating that most of bound fluorescent virions were present extracellularly. Similar experiments using blood mononuclear cells showed that fluorescent HIV-1 particles after binding to these cells were present intracellularly. This flow cytometry data was confirmed in immunogold electron microscopy demonstrating that most HIV 1 gag p24 or FITC label was present at the outside of FDC and on adherent virus particles. We conclude that HIV-1 virions adhere to FDC in vitro in a complement component C3-dependent way. Anti-HIV-1 antibodies in serum from HIV-1 infected patients enhance binding but, by itself, are unable to mediate binding. PMID- 8423333 TI - Appearance of granulocyte-macrophage colony-stimulating factor activity at allergen-challenged cutaneous late-phase reaction sites. AB - The cytokines IL-3 and granulocyte-macrophage CSF (GM-CSF) activate and/or prime monocytes, basophils, and eosinophils for a number of proinflammatory events in vitro. It was hypothesized that IL-3 and GM-CSF might also participate in the local inflammatory cascades that occur at cutaneous blister sites after Ag challenge in vivo. The M-07e megakaryocytic leukemia cell line, which proliferates in response to IL-3 or GM-CSF, was used to determine whether these cytokines were present in fluids derived after Ag challenge in the cutaneous blister chamber model. Fluids from blister chambers after either Ag (timothy grass, orchard grass, or ragweed) or vehicle control challenge were collected hourly for 12 h from nine patients with allergic rhinitis. Cytokine (IL-3/GM-CSF) activity was modestly elevated at 4 h after Ag challenge compared to control with the median of maximal proliferation 4% (range, 2 to 22%) vs 2% (range, 1 to 14%), respectively (Ag vs control, p < 0.03). Activity peaked at 7 h (Ag = 10%, range 1 to 12%, vs control = 1%, range 1 to 9%, p < 0.02) and then steadily declined. No increase in cytokine activity over control was seen in Ag-challenged nonatopics (n = 5, p = NS), indicating that release did not result from a nonspecific effect of the Ag solution. Neutralization of cytokine bioactivity in pooled late phase reaction (LPR) fluids from h 4 to 12 (n = 5) with anti-IL-3, anti-GM-CSF, or both antisera revealed that the majority of the activity was GM-CSF. To better quantify cytokine levels, pooled LPR fluids prepared from an additional 11 subjects were concentration-dialyzed (10x) and tested for cytokine activity. Pooled fluids from Ag-challenged sites contained a median of 625 pg/ml (range, 30 to 1250 pg/ml) GM-CSF equivalents, whereas those from the vehicle control challenged sites contained a median 30 pg/ml (range, 30 to 300 pg/ml) GM-CSF equivalents (p < 0.004 Ag vs control groups, n = 11). Concentrated fluid from Ag- and control-challenged sites in two nonatopic subjects contained < 10 pg/ml cytokine activity. To evaluate the IL-3 and GM-CSF activity with a separate technique, ELISA were performed on separately pooled blister fluids from six atopic subjects. Although no IL-3 activity was detected after Ag challenge in these six subjects, all of them demonstrated levels of GM-CSF at Ag-challenged sites comparable to that found in the bioassay.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423334 TI - Unmodified pancreatic islet allograft rejection results in the preferential expression of certain T cell activation transcripts. AB - Polymerase chain reaction-assisted reverse transcription was used to study the temporal course of rejection in unmodified recipients of murine pancreatic islet cell allografts (DBA/2-->B6AF1) by using syngeneic tissues as controls. The histologic appearance of the grafts was analyzed in parallel. Preproinsulin and constant region of the TCR-beta chain transcripts were studied as markers of graft integrity and infiltrating T cell mass, respectively. The participation of certain cytokines and CTL were analyzed by the detection of IL-2, IFN-gamma, IL 4, and CTL-specific serine protease (granzyme B) transcripts. The time-related disappearance of intragraft preproinsulin transcripts correlated with graft destruction, whereas the intensity of intragraft TCR-beta chain transcript levels correlated with the magnitude of mononuclear leukocyte infiltration in allografts. In unmodified allografts, the magnitude of IL-2 and IFN-gamma intragraft mRNA levels correlated with the intense mononuclear leukocyte infiltrate found on histologic examination at day 8. Only after stable IL-2 gene transcription on day 8 does evidence of graft destruction become apparent, indicating that IL-2 gene activation is closely related to and probably required for expression of alloimmune cytopathic processes. In contrast, IL-4 transcripts were absent or detected in low copy number throughout this time course. Intragraft expression of granzyme B mRNA, a CTL-specific transcript, peaked from day 8 to day 12 in allografts compared with syngeneic grafts or normal tissue. In syngeneic grafts IL-2 and/or IL-4 mRNA was essentially not detected. Although IFN gamma and granzyme B transcripts were detected in syngeneic grafts, after 4 days the levels of detected transcripts were far less than those noted in allografts. In vivo detection of intragraft IL-2 transcripts in the relative absence of detectable IL-4 transcripts strongly suggests IL-2-dependent immune effector mechanisms are associated with, and perhaps responsible, for allograft rejection. Apparently IL-4-dependent effector mechanisms are not necessary for allograft rejection. PMID- 8423335 TI - A monoclonal antibody to recombinant human IFN-alpha receptor inhibits biologic activity of several species of human IFN-alpha, IFN-beta, and IFN-omega. Detection of heterogeneity of the cellular type I IFN receptor. AB - A cDNA encoding a 63-kDa human IFN-alpha R (hIFN-alpha R) has recently been cloned. Mouse cells transfected with this cDNA failed to show any signal transmission for IFN-beta, suggesting the involvement of additional chains in the receptor complex. We have expressed in Escherichia coli and COS 7 cells a soluble recombinant protein (sIFN-alpha R) comprising the extracellular domain of the hIFN-alpha R fused at the carboxyl terminus to a sequence of five histidine residues. The sIFN-alpha R was affinity purified and used to generate mAb. One mAb, 64G12, which recognized the cellular receptor as well as the recombinant proteins was found to neutralize the biologic activity of IFN-alpha, -beta, and omega and natural type I IFN, but not IFN-gamma. To our knowledge, this is the first report of a neutralizing mAb against the human IFN-alpha R. Interestingly, we found that the affinity of this antibody for the cellular receptor depended on the cell line used. These results provide strong evidence that the cloned hIFN alpha R chain is required for binding and signal transmission of all subspecies of type I IFN and suggest a structural heterogeneity of the receptor at the cell surface. PMID- 8423336 TI - Regulation of IFN-gamma expression in vivo. IFN-gamma up-regulates expression of its mRNA in normal and lipopolysaccharide-stimulated mice. AB - Immune responses to such stimuli as tissue injury, infection, and allografting result in localized IFN-gamma expression. Autoregulation of cytokine expression has been described for some cytokines in vitro, but whether this occurs in vivo is unknown. We therefore examined the role of murine IFN-gamma in the regulation of its own expression in vivo after stimulation with bacterial LPS. This agent is known to induce IFN-gamma expression in both spleen and kidney in a T cell independent, cyclosporine-sensitive manner. We found that concomitant administration of a neutralizing mAb to IFN-gamma inhibited not only the MHC expression induced by LPS but also the increased IFN-gamma mRNA expression, suggesting an autoregulatory role for IFN-gamma. Inhibition was dose dependent and observed in both spleen and kidney. The effect was not seen with a neutralizing anti-IL-3 mAb, demonstrating specificity. The inhibition of IFN gamma mRNA expression by the anti-IFN-gamma mAb occurred in both T cell-deficient athymic nude mice and their normal controls, suggesting that the autoamplification of IFN-gamma mRNA in vivo is T cell independent. Administration of rIFN-gamma to unstimulated mice induced IFN-gamma mRNA expression in spleen and kidney, supporting the conclusion that IFN-gamma up-regulates expression of its mRNA. Exposure of resting murine splenocytes to concentrations of rIFN-gamma as low as 10 U/ml in vitro induced expression of IFN-gamma mRNA. Thus, in vivo IFN-gamma may participate in an autoregulatory loop to amplify the amount of IFN gamma expressed both at the site of local inflammation and at remote sites. This would have relevance in the mechanism by which the host defends itself against and prevents dissemination of an infectious agent. PMID- 8423337 TI - IFN-alpha induces homotypic adhesion and Leu-13 expression in human B lymphoid cells. AB - IFN-alpha influences the recirculation and growth of normal and malignant B lymphocytes, although the mechanisms involved are not currently known. Lymphocyte recirculation is fundamentally dependent on cell-to-cell interactions that are mediated by cell surface adhesion molecules. In this report, we examined the relationship between the effect of IFN-alpha on cell-to-cell adhesion processes and induction of the Leu-13 cell surface protein in established human Daudi B lymphoid cell lines that are either sensitive or resistant to the antiproliferative activity of IFN-alpha. IFN-alpha directly triggered homotypic adhesion of IFN-sensitive Daudi B cells in a time- and dose-dependent manner. In contrast, IFN-alpha had no effect on the cell-to-cell adhesion of IFN-resistant Daudi B cells. The capacity of IFN-alpha to trigger homotypic aggregation correlated directly with the level of induction of the cell surface protein Leu 13 and could be potentiated by anti-Leu-13 mAb. Other cytokines also known to influence the proliferation, differentiation, or recirculation of B lymphocytes such as IFN-gamma, IL-2, IL-4, IL-6, TNF-alpha, and low molecular weight B cell growth factor did not induce either Leu-13 expression or homotypic aggregation of Daudi B cells. The adhesion pathway triggered by the IFN-inducible protein Leu-13 required metabolic energy and an intact cytoskeleton but was not dependent on: 1) new protein synthesis; 2) protein kinase C, protein kinase A, or tyrosine kinase activities; or 3) the function of known adhesion molecules including LFA-1, ICAM 1, CD44, or VLA-4. Taken together, these studies demonstrate a fundamental role for IFN-alpha and the IFN-inducible protein Leu-13 in regulating a novel homotypic adhesion pathway in B lymphocytes, and provide insight into the possible mechanisms by which IFN-alpha regulates biologic processes including recirculation. PMID- 8423338 TI - Role of peripheral hemopoietic chimerism in achieving donor-specific tolerance in adult mice. AB - The role of peripheral hemopoietic chimerism in the induction and maintenance of donor-specific tolerance was investigated by our tolerance-inducing method using cyclophosphamide (CP). As has been previously reported, CP injection at a dose of 200 mg/kg to C3H (Thy-1.2, Mls-1b) mice 2 days after priming with 10(8) viable AKR (Thy-1.1, Mls-1a) spleen cells (SC) resulted in both establishment of mixed chimerism and selective elimination of V beta 6+CD4+ T cells in the periphery. When, instead of viable SC, 1300 rad irradiated 10(8) AKR SC were used for priming to C3H mice, CP treatment 2 days after the priming also caused significant but, as compared with priming with nonirradiated viable cells, incomplete elimination of V beta 6+ T cells in the periphery. In these mice, no hemopoietic chimerism was found. In parallel with this incomplete elimination of peripheral V beta 6+ T cells, LN cells of these mice showed reduced but considerable response to AKR SC. However, once hemopoietic chimerism was introduced to these incompletely tolerant mice by an injection with donor-type viable [AKR x C3H]F1 SC 2 days after CP-treatment, LN cells from these newly established chimeras, irrespective of presence or absence of the thymus, became completely nonresponsive to AKR while preserving normal response to BALB/c (third party). This state of nonresponsiveness was accompanied by clonal elimination of the remaining V beta 6+ T cells in the periphery. These results indicate that peripheral chimerism promoted profound tolerance to donor-Mls Ag specifically. Furthermore, from experiments of skin grafting, we demonstrated that tolerance to minor histocompatibility Ag was also achieved in the presence of peripheral hemopoietic chimerism. PMID- 8423339 TI - The fate of anergic T cells in vivo. AB - The kinetics of Staphylococcal enterotoxin B-(SEB) induced peripheral tolerance has been investigated. Ten days after SEB injection, thymectomized BALB/cByJ mice showed suppressed spleen cell proliferative responses to SEB. After 2 mo the SEB specific response was partly recovered. Four months later the response of spleen cells of SEB-primed mice was comparable to those of control mice. The proportion of CD4+, V beta 8+ T cells was diminished in the tolerized mice and was not restored even after the response was recovered. Purified CD4+, V beta 8+ T cells from SEB-primed mice after 4 mo responded similarly to SEB as control CD4+, V beta 8+ T cells. These expressed a similar profile of surface markers compared with that of unprimed control cells, except a homing receptor was slightly lower. An experiment that addressed the possibility that non-anergic T cells expand over time and are in fact responsible for the recovery of the proliferative response showed that such events unlikely occur in vivo. Therefore, the data indicate that T cell anergy is reversible in vivo. It is also suggested that the challenge with superantigen results in neither clonal expansion nor specific CD4+, V beta 8+ T cell memory. PMID- 8423340 TI - Detection and quantitation of cells secreting IL-6 under physiologic conditions in BALB/c mice. AB - IL-6 is a pleiotropic cytokine involved in the regulation of hematopoiesis and lymphocyte activation. An extremely sensitive ELIspot assay was developed to detect and enumerate individual cells secreting IL-6 in normal BALB/c mice. Under physiologic conditions, the frequency of cells producing IL-6 in the bone marrow, spleen, and mesenteric lymph nodes of normal adult animals was approximately 0.5, 0.1, and 0.01%, respectively. FACS analysis revealed that macrophages and Mac1- B220- cells accounted for the vast majority of IL-6 production in the bone marrow, whereas T and B lymphocytes accounted for 4% and 38% of IL-6 production in the spleen. Simultaneous studies comparing cell number with IL-6 levels indicated that the average activated cell produced approximately 85 molecules of IL-6/s. PMID- 8423341 TI - Enhancement of natural killer activity by an antibody to CD44. AB - In this study, we examined the in vitro effect of an anti-CD44 mAb, S5, on NK function using canine PBMC as effectors. S5 enhanced NK activity in a dose dependent and rapid fashion as did IM7, another anti-CD44 mAb that recognizes a common epitope(s) on CD44. Other anti-CD44 mAb (Hermes-1 and S3) that recognize epitopes distinct from S5 and IM7 had a variable effect on NK activity. The activation of increased killing by S5 only occurred when NK-sensitive targets were used, suggesting that lymphokine-activated killer cells were not being induced. Antibody-dependent cell cytotoxicity was not the mechanism involved in the augmentation of NK activity, nor was it Fc receptor-dependent, inasmuch as S5 F(ab')2 was able to increase NK activity. F(ab') fragments of S5 were equivalent to intact S5 in their ability to stimulate NK activity, demonstrating that cross linking of CD44 was not a necessary component of stimulation, and that nonspecific agglutination of target and effector cells was not occurring via the two F(ab) arms. The enhancement of NK function was monocyte-independent and mediated by radioresistant cells, indicating that the antibody enhanced NK cells directly. This finding would suggest that CD44 can direct a transmembrane signal for NK cell activation. PMID- 8423342 TI - T cell-induced suppression of IgG2ab expression in vivo leads to a large reduction of C gamma 2ab mRNA levels. AB - In the T cell-induced suppression of IgG2ab expression, the level at which B cells are blocked in their development to IgG2ab-producing plasma cells was investigated. Although IgG2ab+ lymphocytes were barely detected in normal and IgG2ab-suppressed mice, intracellular IgG2ab was only detected in crude cell extracts from normal mice. B lymphocytes producing IgG2ab were revealed in T cell depleted splenocytes from normal mice (86 +/- 15/10(6) cells), whereas corresponding cell preparations from IgG2ab-suppressed mice were completely free of such lymphocytes. However, in vitro stimulation of cell preparations from both normal and IgG2ab-suppressed mice with LPS plus rIFN-gamma resulted in IgG2ab production. Accounting for differences in spleen size between the two types of mice, these stimuli induced comparable cell proliferation and numbers of IgG2ab producing lymphocytes. In addition, the level of IgG2ab production per cell was similar in the two types of stimulated cells. This demonstrates that normal and IgG2ab-suppressed mice have the same potential to generate IgG2ab-producing cells. By using a sensitive and specific RNase protection assay, C gamma 2ab transcripts were detected in total RNA preparations from IgG2ab-suppressed mice. The levels of C gamma 2ab gene expression in spleen were much lower (between 150 and 400 times less) in IgG2ab-suppressed mice than in normal mice. Taken together, our data suggest that B lymphocytes committed to IgG2ab production represent the target of CD8+ T cells, which we have previously shown to be required for suppression. The target B cells are very efficiently and rapidly silenced, as demonstrated by the absence of detectable serum IgG2ab and corresponding low levels of C gamma 2ab mRNA. PMID- 8423343 TI - Molecular structure of a cross-reactive idiotype on autoantibodies recognizing parenchymal self. AB - The autoimmune B cell repertoire in anti-tubular basement membrane (alpha TBM) disease is focused on the nephritogenic P1 domain of the 3M-1 target Ag in kidney and normally expresses a disease-modifying cross-reactive Id (IdX). The molecular structure of this Id was determined from a library of rat mAb alpha TBM/alpha 3M 1 by using anchor polymerase chain reactions. Our findings provide the first alignment of V region sequences for rat IgG and reveal that specificity for the P1 domain among alpha 3M-1 antibodies is derived from several recurring germ-line VH genes which have not undergone somatic mutation. The IdX in this repertoire localizes to the H chain on Western blot, and to the CDR3 region as deduced from the cDNA encoding several informative clones. Computer modeling of the Id reveals a conformational structure largely dependent on hydroxyl groups in or near turn position 4 of the H chain CDR3 region. These findings demonstrate that regulatory elements protective of autoimmunity are not encoded in the germ line as IdX, but rather emerge from a recombinatorial diversity engaged by the recognition of parenchymal self. PMID- 8423344 TI - Lack of promiscuity in autoantigen-specific H and L chain combinations as revealed by human H and L chain "roulette". AB - Individual H or L chains from a human autoantibody were used to search for other L or H chains that could form antigen-binding fragments, Fab, with the same specificity. The parent Fab (SP1.2) exhibits high affinity binding for thyroid peroxidase (TPO), a 107-kDa protein that is the major autoantigen in human autoimmune thyroiditis. This autoantibody "roulette," performed by using Ig H and L chain gene libraries expressed in bacteria, increased the frequency of TPO binding clones in the new libraries. However, the frequency was still much lower than would be the case if promiscuous combinations with a variety of H or L chains were compatible with specific Ag binding. Nucleotide sequence analysis of the H and L chains of the new TPO-binding clones revealed even more restriction. Thus, with the SP1.2 H chain, all 11 new Fab utilized L chains from the same V kappa 1 family germline gene as SP1.2 itself. Similarly, five of six H chains "captured" by the SP1.2 L chain were very closely related to the SP1.2 H chain. However, one totally different H chain was isolated: SP4.6 has a VH region that differs substantially from that of SP1.2. SP4.6 also has a distinct D region, uses a different JH, and, unlike SP1.2, which is an IgG1, belongs to subclass IgG4. The affinities for TPO of SP4.6 (with the different H chain) and SP1.20 (which had the least mutated L chain germline gene) were similar to that of SP1.2 (approximately 10(-10) M). As expected, the SP1.2 and SP1.20 Fab, which have the same H chain and closely related L chains, bound to the same domain on TPO. However, a similar domain on TPO was recognized by both SP4.6 and SP1.2, despite the fact that their V, D, and J regions are quite different. This observation raises the possibility that the L chain is critical in defining epitope specificity, even in the presence of completely different D regions and nonidentical VH regions. PMID- 8423345 TI - Cytokine secretion by genetically modified nonimmunogenic murine fibrosarcoma. Tumor inhibition by IL-2 but not tumor necrosis factor. AB - Recent investigations have demonstrated that the in vivo growth of weakly immunogenic murine tumors can be inhibited by genetic manipulations that enable them to secrete a variety of cytokines. Inasmuch as most human tumors fail to elicit a detectable host immune response we questioned whether the growth of a nonimmunogenic murine tumor could be inhibited by the secretion of cytokines. We have thus inserted the cDNA encoding for human IL-2 or TNF into the nonimmunogenic murine fibrosarcoma MCA 102. Tumor cells secreting IL-2 failed to grow in vivo despite normal in vitro growth. This growth inhibition required an intact immune system as tumors grew progressively in mice sublethally irradiated before tumor injection. Tumor inhibition was abrogated by the in vivo depletion, by specific mAb before tumor injection, of either CD8+ T cells or NK cells, but not CD4+ T cells. IL-2 secretion by tumor afforded a significant survival benefit to the animal, and IL-2-secreting tumor limited the growth of admixed nonsecreting parental tumor. Histologic evidence and FACS analyses revealed a dense lymphocytic infiltration of IL-2-secreting tumors composed of both CD4+ and CD8+ T cells. In contrast, secretion of TNF failed to inhibit the growth of MCA 102, and similar lymphocyte subset depletions, or administration of specific anti TNF mAb had no effect on the growth of TNF secreting MCA 102. In summary, these investigations demonstrated that the host response to this nonimmunogenic tumor can be markedly enhanced by the genetic manipulation of the tumor cells to secrete IL-2, but not TNF. This strategy has potential application for the development of immunotherapies for nonimmunogenic tumors. PMID- 8423346 TI - The fate of Borrelia burgdorferi, the agent for Lyme disease, in mouse macrophages. Destruction, survival, recovery. AB - The macrophage is a known reservoir for a number of infectious agents, and is therefore a likely candidate site for persistence of Borrelia burgdorferi, the Lyme spirochete. We report that unopsonized B. burgdorferi enter macrophages rapidly, resulting mainly in degradation but occasionally in apparent intracellular persistence. We studied uptake of spirochetes by macrophages by simultaneously labeling infected cells with antibodies to B. burgdorferi and with sequential components of the endocytic pathway, and we examined optical sections (0.5-1.0 micron in thickness) of these cells by confocal fluorescence microscopy at multiple time points after infection. We found that only 5 min of incubation at 37 degrees C were required for nearly 100% of B. burgdorferi to enter a lysosomal glycoprotein-positive compartment, whereas 60 min were required for 90% of the spirochetes to appear in a cathepsin L-positive compartment under the same conditions. We also labeled infected living cells with acridine orange to distinguish live from killed intracellular organisms. Although the large majority of spirochetes within a given cell were dead, we saw occasional live ones up to 24 h (the longest interval examined) after all extracellular organisms had been lysed in distilled water. Moreover, we can reculture spirochetes from macrophages after infection. Persistence of spirochetes within macrophages provides a possible pathogenetic mechanism for chronic or recurrent Lyme disease in man. PMID- 8423347 TI - Characterization of responses of normal human T cells to Trypanosoma cruzi antigens. AB - The stimulation of normal human PBMC by Trypanosoma cruzi Ag was analyzed. PBMC showed significant in vitro proliferation in response to parasite lysate (Tct), with stimulation indices ranging from 10 to 400, peaking at 6 to 7 days. The cells stimulated with Tct produced significant levels of IL-2. To determine which cells proliferated in response to Tct, PBMC were separated into T- and B-enriched cell populations. Purified T cells, but not B cells, proliferated strongly to Tct. The T cell response required APC and was processing dependent. T cell lines generated against Tct proliferated in response to parasite lysate only in the presence of autologous APC and produced IL-2, IL-6, and IFN-gamma but not IL-4 in response to PMA plus ionomycin. Although there were a significant number of CD45Ra+ cells, the majority of the cells in these T cell lines were CD45Ro+. The V beta usage of Tct-responding T cells was heterogeneous, with most V beta genes represented among the responding cells. An immunodominant repeat Ag (TcD) and a ribosomal phosphoprotein (P0) of T. cruzi elicited strong proliferative responses in all subjects tested. These data indicate the presence of T cell-stimulatory Ag in Tct, characterized by nonpreferential usage of the V beta gene families. The strong stimulation of normal human PBMC by Tct may contribute to immunologic alterations seen in T. cruzi infection. PMID- 8423348 TI - Schistosoma mansoni 28-kDa glutathione S-transferase and immunity against parasite fecundity and egg viability. Role of the amino- and carboxyl-terminal domains. AB - We have previously shown that a mAb that inhibits the enzymatic activity of the Schistosoma mansoni 28-kDa glutathione S-transferase (Sm28 GST) also reduces female worm fecundity and egg viability in vivo and in vitro. By peptidic epitope mapping and an activity reconstitution assay, the carboxyl terminus (CT) amino acid residues 190-211 and to a lesser extent the truncated amino terminus (NT) residues 10-43 of the enzyme were identified as mAb recognition sites. Sera from rats immunized with the NT (10-43) and CT (190-211) peptides showed a partial inhibitory effect on Sm28 GST activity in a late phase (6 to 7 wk) but not in an early phase (2 to 4 wk) after immunization. Passive transfer of Sm28 GST inhibiting anti-N- and C-terminal sera, but not of the noninhibitory sera, protected the infected mice by reducing tissue egg deposition and the ability of eggs to hatch. In active immunization experiments, the CT peptide significantly decreased the worm burden (37 to 40%) in mice as did the rSm28 GST (28 to 52%). In terms of tissue egg deposition and egg-hatching ability, immunization with both the NT and CT peptides reproduced the reduction observed after immunization with rSm28 GST. A constant reduction in egg numbers was noted in the small intestines and the livers of the immunized mice. A clear reduction in the ability of intestinal or hepatic eggs to hatch was observed. The results are discussed in terms of the conformational participation of the NT and CT of Sm28 in the expression of GST activity. PMID- 8423349 TI - Modulation of IL-1-induced neutrophil migration by dexamethasone and lipocortin 1. AB - IL-1 is a pro-inflammatory cytokine which controls many features of the immune and inflammatory response. When injected into a mouse 6-day-old air-pouch, human rIL-1 (1 to 100 ng) induced in a dose-dependent fashion a migration of PMN that could be reliably assessed 4 h after injection. Both IL-1 alpha and IL-1 beta were active in this model. The effect of the cytokine was inhibited by local administration of actinomycin D (1 to 10 micrograms), alpha-melanocyte stimulating hormone (200 micrograms), and a mAb recognizing IL-1R type I (10 micrograms). Indomethacin (1 mg/kg), an inhibitor of cyclo-oxygenase, and BW4AC (2 mg/kg), a selective lipoxygenase inhibitor, were without effect but moderate inhibition was seen with the platelet-activating factor antagonist WEB2086 (1 to 10 mg/kg). The glucocorticoid dexamethasone (0.015 to 1.5 mg/kg) potently inhibited the elicitation of neutrophils induced by IL-1 when given systemically 2 h before the cytokine. The steroid-induced anti-inflammatory protein lipocortin 1 (LC1) also produced a dose-dependent inhibition of PMN migration into the pouch with an ED50 of approximately 0.15 to 0.21 mg/kg. The denatured protein was without effect. Passive immunization of mice with a polyclonal sheep antiserum or a mAb raised against LC1 abolished the inhibitory action of dexamethasone whereas preimmune serum or control IgG were without significant effect. These findings provide further evidence that LC1 is involved in the anti-inflammatory action of glucocorticosteroids and suggest that this protein may act as an endogenous regulator of IL-1 action. PMID- 8423350 TI - Methods for assessing complement activation in the clinical immunology laboratory. AB - Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation only indirectly, are insensitive and open to influence by factors other than complement conversion. New approaches to evaluate complement activation directly using sensitive techniques have been developed, and several could be adopted easily in most laboratories. These concentrate on the detection of activation fragments, neoantigens or complexes that only arise as a direct result of complement activation. The wide application of these techniques in research and clinical practice would enhance our understanding of the pathogenesis of a range of inflammatory and infectious diseases. PMID- 8423351 TI - A modification of an affinity procedure for purification of human C1- inhibitor that provides a homogeneous stable preparation. AB - Human C1- inhibitor can be rapidly purified by the affinity chromatography procedure described by Pilatte and his associates (1989), but the inhibitor so purified breaks down during storage or is in a cleaved form when initially purified. By adding an ion-exchange chromatography procedure after the affinity chromatography, a stable, single species of C1- inhibitor molecules is obtained. It is likely that serine proteinases in trace amounts, which may be complexed with some of the C1- inhibitor, are removed during the ion-exchange procedure. This procedure provides a highly purified and useful preparation of C1- inhibitor. PMID- 8423352 TI - Standardization of in vitro synthesis and detection of HIV-1-specific antibodies. AB - Optimal conditions for in vitro anti-human immunodeficiency virus type 1 (HIV-1) antibody (Ab) synthesis and detection were re-appraised. Western blot (WB) and radioimmunoassay (RIA) could detect about 1 and 10 ng, respectively, of HIV-1 specific Ab (HIV-Ab), while the sensitivity of an enzyme-linked immunosorbent assay (ELISA) was much lower. Optimal HIV-Ab recovery was obtained by culturing 2.5 x 10(6) peripheral blood mononuclear cells (PBMC)/ml from seropositive subjects for 16 days in the absence of mitogens; at higher cell concentrations, background levels were unacceptably high. The background of non-de novo synthesized HIV-Ab was due to insufficient PBMC washing and/or cytophilic immunoglobulin (Ig); a particular washing procedure, as well as 24 h peripheral blood mononuclear cells (PBMC) pre-culture, might help in limiting this phenomenon. However, results should be compared with those obtained in cultures containing puromycin especially in infants, where a higher CD16 antigen expression in lymphocytes is likely responsible for increased amounts of cytophilic Ig released in culture supernatants, compared to adults. PMID- 8423353 TI - New immunoassay technique using antibody immobilized on a membrane and a flow cuvette as reaction vessel. AB - Immunoenzymatic detection systems have been developed using human IgG as a model antigen. A membrane with covalently immobilized specific antibodies was placed into a specially constructed ultranarrow flow cuvette and solutions containing the antigen and antibody-peroxidase conjugate were then successively passed through the flow capillary cell. After washing, the membrane was placed into the substrate solution and the intensity of developed colour on the membrane was recorded visually or by a reflection spectrophotometer. The lower detection limit was about 5 x 10(-11) M and the overall analysis time was 10 min. Photoimmobilization was used to immobilize the antibody and thereby permitting control of the protein surface concentration on the membrane as well as the dimensions and shape of the activated region. PMID- 8423354 TI - A simple quantitative in vitro assay for thymocyte adhesion to thymic epithelial cells using a fluorescein diacetate. AB - Recent data verified the role of thymocyte adhesion to thymic epithelial cells (TEC) in T cell development. In order to measure this cellular event as one of routine examinations in immunological studies, a simple quantitative assay in in vitro setting is required. Labeling thymocytes with fluorescein diacetate (FDA) was used to measure quickly and with certainty the adhesion ability of BALB/c thymocytes to TEC cells, a thymic epithelial cell line derived from BALB/c mouse thymic stroma. Thymocytes stained with FDA at the concentration of 2.5 micrograms/ml gave a strong fluorescent intensity (FI) easily detectable in a spectrophotometer and showed the same phenotypical and functional features as unstained cells. As it was confirmed that the cell number correlated well with FI, the number of cells adhering to TEC in wells of a 96-well microplate could be estimated. The major advantage of using FDA was that it took only a few seconds to measure FI of one sample. In contrast, to count the number of adherent cells with conventional methods using a light microscope or radioisotopes required much longer time and care to avoid radiological hazards. Moreover, cell viability and FI of FDA-labeled thymocytes changed little for 24 h, and lysates of FDA-positive cells demonstrated the same FI as living cells. These results indicate that many samples could be applied for FI measurement at the same time after in vivo or in vitro experiments. PMID- 8423355 TI - An immunoassay for the rapid and specific detection of three sialidase-producing clostridia causing gas gangrene. AB - A rapid and methodologically unusual diagnostic test was developed for the specific detection of Clostridium perfringens, C. septicum and C. sordellii, which cause clostridial myonecrosis. Sialidases (EC 3.2.1.18) secreted by these bacterial species were bound to polyclonal antibodies raised against the respective enzyme and immobilized onto microtiter plates. The activity of bound sialidase was determined with the fluorogenic substrate 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid. The assay permits the detection of a minimum sialidase activity of about 0.1-1 mU/ml of sample solution within 2 h. The sensitivity of the test was reduced by about three-fold for sialidase activities in samples containing 50% serum. Only a few, low cross-reactivities, which never exceeded 10% of the homologous reaction, were observed with 12 sialidases from other bacterial sources. Clinical isolates of the three clostridial species were analysed by this assay and gave positive signals in the homologous test. The assay for the detection of C. perfringens was applied to nine samples from patients suspected to be suffering from clostridial myonecrosis. There was a high correlation between the results of the immunoassay and the bacteriological analysis of infection. PMID- 8423356 TI - A new portable device for automatic controlled-gradient cryopreservation of blood mononuclear cells. AB - Protection of the functional integrity of mononuclear cells stored in liquid N2 requires careful control of the freezing procedure. Consequently, optimal quality of cryopreserved cells is usually assured by freezing according to a specified time-temperature gradient generated by computer-controlled freezing devices. While such equipment offers large capacity and secures maximum survival and functional integrity of the lymphocytes upon thawing, it is quite costly and strictly stationary. We have previously developed and tested an alternative, manual device for controlled-gradient lymphocyte freezing, which has proved suitable for field conditions. We report here the development and testing of a similar micro-controller regulated device, allowing unattended and automatic controlled-gradient cell freezing. The equipment exploits the temperature gradient present between the liquid N2 surface and the neck in an ordinary liquid N2 refrigerator. The lymphocyte samples are placed in a small elevator, which is moved through the N2 gas phase by a stepper motor. Time and temperature are measured at regular intervals, and the position of the samples adjusted to ensure that the actual measurements closely match encoded ideal values. Results of assays of the functional integrity and phenotypic composition of human mononuclear cells frozen by the new system were comparable to those obtained when using cells frozen by a commercially available, stationary cell-freezing equipment, or fresh autologous cell samples tested in parallel. Furthermore, there was a good correlation between functional and phenotypic data obtained using frozen and autologous fresh samples of mononuclear cells. The equipment described is low weight and has low N2 consumption, and is thus suitable for the collection and cryopreservation of lymphocytes under field conditions. Furthermore, the technique provides an inexpensive alternative for researchers with a limited requirement for the simultaneous freezing of large quantities of cells. PMID- 8423357 TI - Characterization of morphine-specific monoclonal antibodies showing minimal cross reactivity with codeine. AB - Six murine monoclonal antibodies against morphine were produced using N-(4 aminobutyl)normorphine as a hapten. Most of the antibodies obtained distinguished the substituents at the 3 and 6 positions of morphine. This property of the antibodies led to a reduction in cross-reactivity with codeine, morphine-3 glucuronide (M-3-G) and morphine-6-glucuronide (M-6-G) to negligible levels. However, one of the antibodies distinguished the substituent only at the 3 position of morphine, which cross-reacted with M-6-G, naloxone and naltrexone. In the competitive inhibition enzyme-linked immunosorbent assay, morphine was detected at concentrations as low as circa 100 pg/ml. PMID- 8423358 TI - Enzyme immunoassay for the determination of bovine growth hormone using avidin biotin-peroxidase complexes. AB - The development of a competitive enzyme immunoassay for bovine growth hormone (bGH) is described. Antiserum, raised in rabbits and diluted 1/250,000, was preincubated with samples and free antibodies from the reaction mixture were immobilized using a microtiter plates coated with bGH. Bound antibodies remaining from the preincubation were visualized using a biotinylated second antibody as a bridge for subsequent amplification by an avidin-biotin-peroxidase complex. The measuring range was between 0.5 and 100 ng/ml. Cross-reactivity with other pituitary hormones was < 0.1%. The intra- and interassay coefficients of variations were 8.1 and 12.7%, respectively, and the recovery of added bGH was 110%. To validate the assay, two bull calves were treated with bGH-releasing factor. The response showed that there was an immediate rise in immunoreactive bGH which peaked after 5 and 15 min at 200 ng/ml and 60 ng/ml, respectively. This enzyme immunoassay is an economic and sensitive alternative to the established radioimmunoassay, and the first competitive enzyme immunoassay described for bGH. PMID- 8423359 TI - Determining confidence limits for drug potency in immunoassay. AB - Nonlinear models have frequently been used to characterize dose-response data obtained from biological assays. The effect of a bioactive agent is observed and the model allows prediction of the dose required to obtain the observed effect (the 'inverse prediction'). The precision of this estimate is important in potency determination. Here, a general method is presented for calculating the inverse confidence intervals for estimates of dose potencies obtained from nonlinear models often used to describe these tests. The approach is demonstrated with application of data sets to the negative exponential and four-parameter logistic regression models. Necessary theory is presented and followed by detailed discussion in which estimation strategies are explained and intermediate quantities calculated. PMID- 8423360 TI - A novel method for controlling the pepsin digestion of antibodies. AB - We have discovered that 0.5-0.8 M ammonium sulfate has a beneficial effect on the peptic digestion of several antibodies. Initially, the project was begun to address the troublesome precipitation observed when pepsin was used to digest monoclonal antibody CYT-368 (C46), an IgG2a that binds carcinoembryonic antigen. High concentrations of various salts effectively blocked this precipitation, and also accelerated the rate of digestion. Of these, ammonium sulfate had the greatest effect. Therefore, it was tested on the peptic digestion of ten other murine IgG antibodies as well as rabbit, sheep and goat immunoglobulins. Besides preventing precipitation, ammonium sulfate usefully modulated the digestion rate, accelerating the digestion of some antibodies and reducing it for others. Finally, ammonium sulfate altered the specificity of pepsin during the digestion of antibody CYT-099 (B72.3) preventing the enzyme from cleaving in the F(ab')2 region, and thereby preserving the activity of the fragment. With ammonium sulfate we were able for the first time to use pepsin to obtain active F(ab')2 in good yield from this widely used antibody. PMID- 8423361 TI - Locomotion through three-dimensional type I rat tail collagen. A modified mini assay. AB - There is an increased application of three-dimensional type I rat tail collagen as an in vitro model for the peritumoral matrix in analysis of lymphocyte migration. The increased demand prompted us to modify the previous methods. We here describe our 'mini'-setup of the collagen model assay, which uses only 1/20 the amount of collagen medium and the number of cells used in the conventional assay. The modified assay was tested for optimal collagen concentration in gel for upward and downward migration, for locomotion from a collagen-gel bead into a collagen overlayer for demonstration of the effect of inhibitors and for differentiation between locomotory properties of lymphocyte subpopulations. The results verify that the mini-assay is an applicable in vitro model, easily read and amenable to limited blood samples such as those obtained from cancer patients, and reflects well known in vivo events. PMID- 8423362 TI - Development of a latex agglutination inhibition reaction test for amphetamines in urine. AB - A simple and rapid immunoassay of amphetamines based on latex agglutination inhibition reaction has been developed. N-(3-aminopropyl) amphetamine, a novel amphetamine derivative, and its BSA conjugate were synthesized and characterized. The hapten density in the conjugate was determined spectroscopically to be 62.52 mol/mol of BSA. Two other immunogens, amphetamine-BSA and amphetamino succinamide BSA, were also synthesized and studied. It was found that N-(3-aminopropyl) amphetamine-BSA exhibits favorable features in terms of immunogenicity and immunochemical specificity when compared to the other two amphetamine immunogens. A latex agglutination inhibition reaction test (LAIRT) using DEAE-cellulose purified rabbit IgG against N-(3-aminopropyl) amphetamine-BSA was found to give a sensitivity of 0.6 microgram/ml and 4.0 microgram/ml of amphetamine and metamphetamine, respectively. Various commonly used drugs and narcotics at concentrations 0.8 mg/ml or less, did not interfere with the test. Interference by normal urine was observed but it could be eliminated by the inclusion of 0.78% normal rabbit serum. The sensitized latex was stable at 4 degrees C for at least 6 months. It was also stable to lyophilization and to at least four cycles of freezing and thawing. The total test time was 35 min. Comparison was made between the LAIRT and EMIT-d.a.u. on 56 urine samples collected from truck drivers. While the EMIT showed 47 positives and nine negatives, the LAIRT gave 38 positives and 18 negatives. The two tests showed no statistical significant difference (P < 0.05). PMID- 8423363 TI - A time-resolved fluoroimmunoassay for human insulin based on two monoclonal antibodies. AB - The insulin time-resolved fluoroimmunoassay (TRFIA) described here is a 'sandwich type' immunoassay using two monoclonal antibodies recognizing different epitopes of insulin. One of the antibodies is fixed to a solid phase and the other is biotinylated. After sandwich formation the biotinylated antibody is detected by time-resolved fluorometry with the fluorophore europium which is coupled to streptavidin. Serum samples of 100 microliters can be analyzed within a concentration range of 4 microIU/ml up to 500 microIU/ml. The detection limit of this assay, 3.9 microIU/ml, is comparable to the conventional radioimmunoassay and the results overall correlate well with those of the radioimmunoassay (r = 0.99). There is no detectable crossreaction with human proinsulin even using a proinsulin concentration of 2000 microIU/ml. The assay is performed with the DELFIA system (Pharmacia). PMID- 8423364 TI - An improved MTT assay. AB - The MTT assay for cell viability and cell proliferation has been modified to improve its reproducibility and accuracy. The modified test is performed on MultiScreen filtration plates, which permits the removal of the culture medium prior to formazan solubilisation, without loss of cells or formazan crystals. A 1:1 mix of DMSO and ethanol is used as the solvent, since this has the same optical refraction index as the filters, making it possible to measure the optical densities directly on the MultiScreen plate. Data obtained in the assay method using the MultiScreen plate were compared with data from studies employing the normal flat-bottomed plate, by using cells which grow in suspension. Two cell lines were used in the study. CTLL-2, which are IL-2 dependent cells of murine T cell origin, and Jurkat E.6.1 which are IL-2 producing cells of human lymphoma origin. CTLL-2 cells and the modified MTT assay were also used for evaluating the effects of different IL-2 concentrations on cell proliferation. PMID- 8423365 TI - Eupergit C-coated membranes as solid support for a sensitive immunoassay of human albumin. AB - A number of commercially available membranes preactivated by coating with oxirane containing Eupergit C beads were used for diagnostic immunoassay applications. These membranes possess a higher capacity for protein binding than the respective unmodified membranes as measured with fluorescently labelled antibodies. Immobilization of antigens or antibodies as the first step of the assay was achieved by covalent binding of amino groups of proteins to the oxirane moieties introduced onto the membranes. The high performance of the newly developed membranes bearing covalently bound antibodies, was demonstrated by a dot enzyme linked microalbuminuria immunoassay as compared to unmodified membranes. PMID- 8423366 TI - SDS-PAGE after micro-affinity adsorption for analysis of heterogeneous antigen polypeptides in individual sera. AB - A method was developed for the analysis of heterogeneity in antigen polypeptides in individual sera. Polypeptides in sera were adsorbed by polystyrene beads coated with antibody in wells of a microplate. They were dissociated with a small volume of elutant, and transferred to slots on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Polypeptides separated on gel were then immunoblotted with antibodies labeled with horseradish peroxidase. The method was applied to analyze different populations of hepatitis B surface and e antigen polypeptides in sera from carriers of hepatitis B virus. Applicability to mass-scale and high sensitivity of the method would allow surveys of heterogeneous antigen polypeptides in serum for any biological significance. PMID- 8423367 TI - Comparative analysis of using MTT and XTT in colorimetric assays for quantitating bovine neutrophil bactericidal activity. AB - Two different tetrazolium compounds were compared for use in a colorimetric assay for quantitating bovine neutrophil bactericidal activity against Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, and Brucella abortus. The tetrazolium compounds tested included 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) and sodium 3,3'-[1[(phenylamino)carbonyl]-3,4- tetrazolium]-bis(4-methoxy-6-nitro) benzene sulfonic acid hydrate (XTT). The MTT and XTT colorimetric bactericidal assays were conducted by incubating antibody opsonized bacteria with neutrophils in microtiter plates for 30 and 60 min at ratios of ten and 100 bacteria per neutrophil. Neutrophils were then lysed with saponin and samples were incubated 30 min with MTT or XTT plus coenzyme Q (CQ). Dead bacteria and lysed neutrophils did not react with MTT or XTT plus CQ. Live bacteria converted XTT to water soluble orange formazan in the presence of CQ and MTT to insoluble purple formazan. Absorption of formazan produced by bacteria from XTT was measured at 450 nm. Formazan produced by bacteria from MTT was solubilized by adding isopropanol and measured by absorption at 560 nm. Absorption of both types of formazan was directly related to viable bacteria cell number and used to determine the number of bacteria not killed by neutrophils. The percentage of bacteria killed by neutrophils was determined by extrapolation from a standard formazan curve that was derived by incubating MTT or XTT plus CQ with known numbers of bacteria. The XTT and MTT colorimetric bactericidal assays produced comparable results when used to measure bovine neutrophil bactericidal activity against S. aureus, E. coli, L. monocytogenes, and B. abortus. However, the assay using XTT was quicker and easier to perform because bacteria converted XTT to a formazan that did not need to be solubilized before measuring absorption. PMID- 8423368 TI - Use of an aqueous soluble tetrazolium/formazan assay to measure viability and proliferation of lymphokine-dependent cell lines. AB - A new tetrazolium compound, MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt), has recently been described which in the presence of phenazine methosulfate (PMS) is reduced by living cells to yield a formazan product that can be assayed colorimetrically. An important advantage of MTS/PMS over other tetrazolium dyes (e.g., MTT) is the aqueous solubility of the reduced formazan product which eliminates the need for detergent solubilization or organic solvent extraction steps. Its advantages over XTT/PMS, another tetrazolium which yields a water-soluble formazan product, include the absorbance range of color produced (515-580 nm as opposed to 450 nm), the rapidity of color development, and the storage stability of the MTS/PMS reagent solution. In the present study, MTS/PMS was used to assay viability and proliferation of the IL-2-dependent HT-2 and CTLL-2 cell lines and the IL-3 dependent FDC-P1 and FL5.12 cell lines. With each cell line, the amount of formazan product was time-dependent and proportional to the number of viable cells. Furthermore, with both HT-2 and CTLL-2 cells it was found that cultures could be simultaneously labeled with MTS/PMS and [3H]thymidine, with relatively little effect of the dye on uptake of the latter. This feature was further capitalized upon in studies with FDC-P1 cells, in which the co-addition of MTS/PMS and [3H]thymidine was used to distinguish between cell viability and proliferation. PMID- 8423369 TI - Dye-ligand affinity purification of human complement factor B and beta 2 glycoprotein I. AB - The rapid purification of human factor B using dye-ligand chromatography is described. The 50% ammonium sulphate supernatant of fresh human serum is equilibrated in pH 7.4, 25 mM Tris buffer containing 0.5 mM CaCl2 and 0.5 mM MgCl2, with 25 mM sodium caprylate and chromatographed on Cibacron Blue F3GA agarose. Caprylic acid binds to the fatty acid binding site of albumin, preventing it from binding to the resin which thus retains a high capacity for binding factor B. Factor B together with the homologous protein beta 2I are eluted from the column by a linear gradient of KCl. Subsequent NaCl gradient FPLC on Hiload S-Sepharose, equilibrated in 10 mM potassium phosphate, 5 mM EDTA, pH 7.0, provides both factor B and beta 2I in homogeneous form. PMID- 8423370 TI - Application of the MAILA technique to the study of human anti-HLA monoclonal antibody specificity. AB - The monoclonal antibody-specific immobilization of lymphocyte antigens (MAILA) assay was developed to detect antibodies present in human alloantisera against antigens of different major histocompatibility complex loci, particularly of class II specificity. The MAILA assay has been used in our laboratory to the determination of the type of HLA molecule recognized by human monoclonal antibodies 91C2 (anti-A2 + 28), 34F11 (anti-DQ1), and 2A2 (anti-DQ1 + 4 + short DQ7), using well characterized monomorphic as well as polymorphic murine monoclonals for the specific immobilization of HLA molecules. Results obtained show that the MAILA assay is also a valuable tool for the determination of specific human MHC locus products recognized by human monoclonal antibodies. PMID- 8423371 TI - Improved sensitivity in the antibody-forming cell assay for Japanese encephalitis virus in mice by optimal fixation of cells and avidin-biotin complex (ABC) immunocytochemistry. AB - A sensitive modified method for the detection in vitro of antibody-forming cells (AFCs) in Japanese encephalitis (JE) virus infected mice is described. The procedure involves the selection of the most appropriate incubation period, after the infection of monolayer cells, for expression of viral antigens to be recognized by AFCs. Adequate preservation of the expressed antigens was achieved after fixation with paraformaldehyde-lysine-periodate buffer (PLP). The development of spots which corresponded to AFCs was successfully obtained with the use of the avidin-biotin complex (ABC) amplification reaction. The number of the spots developing after completion of the assay was greater after PLP fixation compared with other fixation solutions such as methanol, formalin and acetone water. Optimum antigen expression on the infected cells for recognition by lymphoid cells was reached after 24 h of infection. However, it was possible to detect viral antigens on the infected cells 8 h after infection. The ABC reaction was shown to be the best procedure for developing spots compared with other immunocytochemical methods. A linear increment was observed in the number of spots that developed at different spleen cell densities. This assay is simple to perform and could detect virus-specific Ig producing cells from the spleens of mice infected with JE virus. It also makes it possible to enumerate accurately the kinetics of the AFC response in different lymphoid organs during infection or after vaccination. PMID- 8423372 TI - Artifacts with the thiocyanate elution method for estimating relative antibody avidity. PMID- 8423373 TI - Separation of different forms of the fourth component of human complement by fast protein liquid chromatography. AB - Disruption of the thiolester in native C4 yields a 'C4b-like C4' molecule (iC4) that functionally resembles C4b and is therefore probably accompanied by conformational changes in the C4 molecule. In most purified C4 preparations, iC4 and C4b are present to a variable extent. In this study we evaluated the use of fast protein liquid chromatography (FPLC) to resolve and isolate these various forms of C4. C4 was purified from fresh human plasma in a 4-step procedure that included barium citrate adsorption, polyethylene glycol 6000 (PEG) precipitation, Q-Sepharose Fast Flow and mono Q ion exchange chromatography. The final preparation appeared to be homogeneous on SDS-PAGE and under reducing conditions consisted of three bands that corresponded to the intact alpha, beta and gamma chains of C4. In some preparations the alpha' chain of C4b was also observed. On a Mono Q column the purified C4 preparations could be separated into three peaks that by hemolytic assay and SDS-PAGE were characterized as representing native C4, and monomeric and dimeric iC4 (or monomeric and dimeric C4b). Finally, the apparent KA of the various forms of C4 for C4b-binding protein (C4BP) was investigated. The monomeric iC4 and C4b species demonstrated similar C4BP binding affinity with an apparent KA of 5.6-6.4 x 10(8) M-1, whereas their dimeric forms demonstrated a higher affinity for C4BP with an apparent KA: 0.9-2.3 x 10(9) M-1. Binding of native C4 to C4BP was undetectable. PMID- 8423374 TI - A flow cytometric rosetting assay for the analysis of Fc receptors and C3 receptors on HSV-infected cells. AB - A sensitive and reproducible flow cytometric assay was developed for the analysis of Fc gamma and C3b(i) receptors on HSV-infected cells. The method is based on a rosette technique using fluorochrome-labeled erythrocytes sensitized with IgG or C3b(i). A comparison of flow cytometric and microscopic quantitation demonstrated that the binding of EIgG, EC3b(i) to HSV-infected cells were correlated. Flow cytometric analysis provides the opportunity to study simultaneously the distribution of E per HSV-infected cell and the total binding of E to the whole population of HSV-infected cells. Receptor activity and HSV glycoprotein cell surface expression were shown to be correlated in a linear fashion. The assay could be applied to other Fc gamma R- and C3b(i)R-bearing cells. PMID- 8423375 TI - Capture assay for specific IgE. An improved quantitative method. AB - A capture assay for the measurement of specific IgE in the serum of allergic patients is described, using monoclonal anti-human IgE (coated to the wells of a microtiter plate) and biotinylated allergens in solution. In a single incubation, IgE is bound to the solid phase through the Fc fragment and biotinylated allergens react with their specific IgE Fab regions, if present. In a second step, streptavidin-HRP conjugate is added to reveal the amount of biotin fixed on the solid phase. Quantitative determinations are obtained by comparison with a standard curve of total IgE incubated with a biotinylated monoclonal anti-IgE, complementary to the one employed as capture antibody. The assay is unaffected by allergen-specific IgG, shows good intra- and interassay reproducibility and is linear over a wide range of specific IgE concentrations. The method could be used with a wide range of different allergens. PMID- 8423376 TI - An efficient sandwich-ELISA for the determination of choline acetyltransferase. AB - A specific, sensitive, and reliable sandwich-ELISA (enzyme-linked immunosorbent assay) has been established for the determination of choline acetyltransferase (CHAT) from porcine brain. The detection limit of the assay was 30 micrograms/l and the assay was linear up to 300 micrograms/l. The within-day and day-to-day coefficients of variation were found to be 3.3% and 4.7% respectively for low CHAT concentrations (30 micrograms/l) and 3.1% and 3.4% respectively for high levels of CHAT (300 micrograms/l). The immunoassay was more sensitive than the radiometric assay of Fonnum which is widely used for the measurement of enzyme activity. In the assay monoclonal antibody was adsorbed to the polystyrene surface of the immunoplate as the capture reagent. Using a standard peroxidase protocol the immobilized antigen was detected with a highly specific anti-CHAT antiserum raised in rabbits. Two monoclonal antibodies were available for antigen binding. One of the two--A10.29B4--reacted preferentially with the active enzyme the other one--B3.9B3--reacted only with a degraded form. The polyclonal antiserum recognized both native and denatured enzyme. The effectiveness of employing the two monoclonal antibodies separately or in combination was demonstrated by measurement of porcine CHAT diluted in human cerebrospinal fluid and serum. After some minor modifications the sandwich-ELISA could be used for the determination of CHAT from the central nervous system of the rat. PMID- 8423377 TI - Assessment of mycobacterial infection and multiplication in macrophages by polymerase chain reaction. AB - The amplification of mycobacterium-specific DNA sequences from samples obtained from infected patients by the polymerase chain reaction (PCR) has been useful in the clinical diagnosis of mycobacterial diseases. Using 20 bp oligonucleotide primers that recognize a 123 bp repeated sequence present in M. bovis and M. tuberculosis DNA, we describe in detail the conditions of the PCR reaction that allow an assessment of the mycobacterial content of infected macrophages. The results of the highly reproducible, time-efficient PCR technique show good correlation with the widely used colony forming unit (CFU) and [3H]uracil incorporation methods for the detection of Mycobacterium. Our method allows an assessment of the level of M. bovis BCG infection from a variety of sources, including peritoneal macrophages and macrophage lines, within a few hours, making it the assay of choice for rapid determination of the level of mycobacterial growth in infected cells, in experimental models of mycobacterial infection. PMID- 8423378 TI - Connective tissue diseases and the skin. Memorial supplement for James N. Gilliam, M.D. PMID- 8423379 TI - Role of microvascular endothelial cells in inflammation. AB - Endothelial cells are critical elements in the evolution of all types of cutaneous inflammation. They participate through the synthesis and secretion of pro-inflammatory cytokines, including interleukin 1 (IL-1), IL-6, and IL-8, as well as M-CSF, G-CSF, GM-CSF, gro alpha, and MCP. They also express a series of cell-surface proteins and glycoproteins known as cell adhesion molecules that allow circulating leukocytes to bind to endothelial cells and allow endothelial cells to bind to matrix proteins. The regulated expression of these molecules, including those in the integrin, immunoglobulin gene, and selection families, allows for the precise trafficking of circulating leukocytes to sites of inflammation, injury, or immunologic stimulation in the skin. Furthermore, emerging evidence clearly indicates that selected differences exist between endothelial cells of the microvasculature and those that line large blood vessels. These include differences in secreted products, differences in the expression of cell adhesion molecules, and differences in cytokine-induced regulation of commonly expressed cell adhesion molecules, among others. Thus, a precise delineation of the biology of cutaneous microvascular endothelial cells is important to our understanding of cutaneous inflammation. PMID- 8423380 TI - Humoral immunity in polymyositis/dermatomyositis. AB - Autoantibodies are found in most patients with polymyositis (PM) or dermatomyositis (DM) and 35-40% of these patients have myositis-specific antibodies. Twenty-five to thirty percent have anti-aminoacyl-tRNA synthetases, of which anti-Jo-1, directed at histidyl-tRNA synthetase, is by far the most common. Patients with anti-synthetases have a high frequency of myositis, interstitial lung disease, Raynaud's phenomenon, and other features constituting an "anti-synthetase syndrome." Anti-synthetases tend to react with conformational epitopes and to inhibit enzymatic activity, suggesting reaction with conserved regions. Sera with antibodies to alanyl-tRNA synthetase (anti-PL-12) also have antibodies to tRNA(ala), whereas most sera with other anti-synthetases do not react directly with tRNA. Production of the antibodies appears to be antigen driven, and is influenced by HLA genes, although an initiating factor, possibly a viral infection, may be important. Antibodies to other cytoplasmic antigens, most notably the signal recognition particle (anti-SRP), are seen in a small percentage of patients. Patients with anti-SRP do not tend to develop the anti synthetase syndrome, but may have very severe disease. Antibodies to the nuclear antigen Mi-2 are also specific for myositis, and are strongly associated with DM. Several autoantibodies, including anti-PM-Scl, anti-Ku, and anti-U1 and U2 RNP, have been associated with scleroderma-PM overlap. The role of humoral immunity in the myositis of PM and DM has not yet been clarified. Capillary loss and ischemic damage are important in DM, and seem to be mediated by humoral mechanisms, whereas cell-mediated attack on muscle fibers is important in PM. The mechanism of skin injury in cutaneous lesions is not known, but antibody deposition is inconsistent and uncommon. Whether the myositis-specific antibodies are involved in disease pathogenesis is not yet known, although there is no direct evidence for this. An understanding of the reasons for production of these antibodies, however, should provide insight into the etiology and pathogenesis of PM and DM. PMID- 8423381 TI - Amyopathic dermatomyositis: a review. AB - Jim Gilliam's research interests throughout his career were forced upon better defining the relationships that exist between the cutaneous and systemic manifestations of the rheumatic diseases. Although the majority of his time was spent studying such relationships in lupus erythematosus patients, he was also intensely interested in dermatomyositis (DM) in this regard as well. He was particularly intrigued with the dissociation of the cutaneous and muscular manifestations of this disorder that occasionally occurs. The term "dermatomyositis sine myositis" has been used in the past to describe patients who present with only the cutaneous manifestations of DM; however, very little published data is available from systematic examinations of such patients. For several reasons, we have preferred the term "amyopathic dermatomyositis" to describe that rare patient who for long periods of time suffers from the classical skin lesions of DM as the only clinically significant manifestation of their disease. In this presentation, we review our own personal experience with a group of six such patients and compare and contrast it to that of other workers who have dealt with this subject over the past two decades. PMID- 8423382 TI - Dermatomyositis and malignancy. AB - During the past 12 years, many studies applying strict diagnostic criteria have been published that have attempted to settle the controversy about the reality of the association between dermatomyositis and malignancy. Although retrospective, recent studies have shown an increased incidence of malignancy among patients with dermatomyositis when compared with controls without myositis. In contrast, an increased frequency of malignancy in dermatomyositis as compared to polymyositis still has to be demonstrated. In most cases, malignant disease precedes or occurs concurrently with dermatomyositis and is discovered on the basis of clinical signs, symptoms, and routine screening laboratory tests. The types of neoplasms found in association with dermatomyositis parallel those observed in the general population. A possible link between dermatomyositis and an underlying malignancy remains largely hypothetical at a biologic level, although cellular immunity abnormalities may provide a direction for future investigations. Prospective epidemiologic studies using the case-control methods and cohort analysis remain necessary 1) to rigorously demonstrate the reality and to study the nature of the association between dermatomyositis and malignancy, and 2) to clarify the optimal screening strategies for malignant neoplasms in patients with dermatomyositis. PMID- 8423383 TI - Anti-Ro(SS-A) HLA-DR3-positive women: the interrelationship between some ANA negative, SS, SCLE, and NLE mothers and SS/LE overlap female patients. AB - During the past 15 years, the clinical spectrum associated with the anti-Ro(SS-A) antibody response has been defined. Various clinical presentations, including subacute cutaneous lupus erythematosus, the neonatal lupus syndrome, the Sjogren's syndrome/lupus erythematous overlap syndrome, and primary Sjogren's syndrome, have been detected in association with the anti-Ro(SS-A) response. The anti-Ro(SS-A) antibody response is associated with the HLA-DR2 and HLA-DR3 phenotypes. There is now a good deal of evidence to suggest that many anti-Ro(SS A)-positive HLA-DR3 women are genetically closely related, sharing in common an enriched frequency of the HLA-DR3-linked B8, DQw2, and DRW52 phenotypes. DNA sequence studies have confirmed this genetic relationship. These studies have led us to the following conclusions. 1) The HLA-DR2 and HLA-DR3 associations with systemic lupus erythematosus and the HLA-DR3 association with Sjogren's syndrome are related to the anti-Ro(SS-A) antibody response and not to the clinical disease expression. 2) HLA-DR3 anti Ro-positive female patients with first-degree Sjogren's syndrome, subacute cutaneous lupus erythematosus, or Sjogren's syndrome, or who are asymptomatic, are immunogenetically closely related even though the clinical presentations are strikingly different. All these HLA-DR3 anti-Ro(SS-A) antibody-positive women are at risk to give birth to a child with the neonatal lupus syndrome. PMID- 8423384 TI - Major histocompatibility class II antigen expression on the surface of epidermal cells from normal and ultraviolet B irradiated subjects. AB - The influence of ultraviolet B irradiation in therapeutic doses on MHC II positive epidermal cell numbers and their surface MHC II antigen expression was studied quantitatively using light microscopic immunoperoxidase and immunogold electron microscopic techniques. In multiple ultrathin sections through many MHC II-positive epidermal cells from five healthy subjects, prior to ultraviolet exposure, Langerhans cells and indeterminate cells were found to express similar densities of surface MHC II antigens, which were uniformly distributed over the cell surface. The variation in surface MHC II antigen expression on 97 dendritic epidermal cells from one subject was normally distributed. Following a 6-week course of ultraviolet B irradiation, in the same doses as those normally used for the treatment of psoriasis, MHC II-positive epidermal cell numbers were significantly reduced (mean decrease to 51% of the pre-UVB sample; p < 0.001 analysis of variance), but their surface MHC class II antigen density was significantly increased (p < 0.05 analysis of variance). No MHC II-negative Langerhans cells were detected in either the pre- or post-UVB samples. PMID- 8423385 TI - Subacute cutaneous lupus erythematosus. AB - Gilliam recognized subacute cutaneous lupus erythematosus (SCLE) as a lupus specific eruption that identifies a unique subset of lupus erythematosus. These patients were noted to have prominent photoaggravated skin disease and often had musculoskeletal complaints, but generally did not develop significant systemic disease. SCLE patients were later found to have other distinctive features, including the frequent presence of anti-Ro antibodies, and enrichment for the human histocompatibility antigens (HLA) B8 and DR3. In the 13 years of published reports of SCLE patients following the initial study by Sontheimer et al (Subacute cutaneous lupus erythematosus: a cutaneous marker for a distinct lupus erythematosus subset. Arch Dermatol 115:1409-1415, 1979) a number of additional observations regarding SCLE patients have been made. These have included the recognition that SCLE may be associated with other rheumatic diseases, and that photoactive medications may induce lesions of SCLE. Areas of controversy concerning SCLE include conflicting studies regarding the histopathology of SCLE as compared to discoid lupus erythematosus (DLE), as well as the frequency of detection of anti-Ro antibodies in SCLE patients. Recent interesting studies of SCLE include a description of a unique pattern of immunoglobulin G (IgG) deposition on direct immunofluorescence, which may indicate the binding of anti Ro antibodies to keratinocytes in vivo. PMID- 8423386 TI - Antiphospholipid syndrome. AB - The antiphospholipid antibodies (aPL), namely, the lupus anticoagulant and the anticardiolipin antibodies, are a family of autoantibodies directed predominantly against negatively charged phospholipids. Many studies have confirmed that patients with these antibodies are prone to repeated episodes of thrombosis, fetal losses, and thrombocytopenia. The association of aPL with these clinical events has been termed the antiphospholipid syndrome. Several skin lesions have been found in patients with this syndrome, including livedo reticularis, livedoid vasculitis, thrombophlebitis, cutaneous infarctions and gangrene of digits, ulcerations, lesions resembling vasculitis (nodules, macules), cutaneous necrosis/infarctions, subungual splinter hemorrhages, and, less commonly, discoid lupus and Degos' disease (malignant atrophic papulosis). In this article, we review the main immunologic and clinical aspects of this syndrome with special emphasis on the dermatologic features. PMID- 8423387 TI - Cultured human keratinocytes synthesize and secrete endothelin-1. AB - The human epidermal-melanin unit exists as a complex interplay of cell-cell interactions. Melanocytes synthesize melanin and transfer it to the surrounding keratinocytes, which, in turn, produce factors that affect melanocyte homeostasis, growth, and melanization. Endothelin-1 (ET-1), a vasoconstrictor peptide produced by endothelial cells, has recently been shown to stimulate human melanocyte proliferation and tyrosinase activity. To investigate the possibility that keratinocytes synthesize and secrete ET-1, we grew human keratinocytes in a defined serum-free medium and measured ET-1 levels in the keratinocytes and the keratinocyte-conditioned medium. Northern analysis of keratinocyte total RNA also was performed. We found that human keratinocytes express preproET-1 mRNA and translate the message to ET-1 protein, which is secreted into the keratinocyte medium. Human keratinocytes produced ET-1 in a time-dependent manner with total production of 20.1 +/- 1.1 pg ET-1/10(6) cells at 24 h (n = 7). Although total ET 1 production (secreted plus cell-associated ET-1) was similar, the proportion of secreted versus cell-associated ET-1 varied widely among the different donors. We have found that human keratinocytes synthesize and secrete ET-1 in vitro. From these data we believe that the keratinocyte could be an in vivo epidermal source of this melanocyte growth and pigmentation factor. PMID- 8423388 TI - A new 180-kDa dermal endothelial cell activation antigen: in vitro and in situ characteristics. AB - PN-E2 is a monoclonal antibody generated against recombinant tumor necrosis factor-alpha (TNF-alpha)-treated human umbilical vein endothelial cells (EC). PN E2 recognized a molecule with expression levels in vitro that could be downregulated by TNF, and in situ PN-E2 showed only weak reactivity with vascular EC in normal skin, as assessed by immunohistochemical staining. The expression of PN-E2 was considerably increased on EC in various pathologic skin lesions, including psoriasis, granulation tissue, and inflamed skin. PN-E2 antigen expression was analyzed in more detail in vitro on cultured EC and fibroblasts by use of enzyme-linked immunosorbent assay and fluorescence-activated cell sorter techniques. The expression level on human umbilical vein endothelial cells and capillary EC was, in contrast to the in situ immunohistologic findings, invariably high. On fibroblasts, a low expression was found. Incubation of the EC with recombinant TNF-alpha decreased expression by a factor of 2. Incubation of EC with recombinant interferon-gamma resulted in a twofold increase in PN-E2 antigen expression, whereas other cytokines [recombinant interleukin (rIL)-1 alpha, rIL-1 beta, rIL-4, rIL-6], lipopolysaccharide, or recombinant basic fibroblast growth factor had no effect. Immunoelectron microscopy of tissue specimens and EC preparations localized the antigen on the luminal membrane of the endothelium. Immunoprecipitation followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed a major band at 90 kDa and a minor band at 80 kDa under reducing conditions and bands of 180 and 400 kDa under non-reducing conditions. Molecular weight and expression patterns in vitro on EC after incubation with cytokines excluded most of the known endothelium-specific molecules, with the possible exception of endoglin (the 44G4 antigen). We conclude from our findings that this new antigen could be useful as a marker for endothelial activation in skin biopsy material. PMID- 8423389 TI - Bullous SLE: a phenotypically distinctive but immunologically heterogeneous bullous disorder. AB - Bullous systemic lupus erythematosus (SLE) is a rare blistering disease with a distinctive combination of clinical, histologic and immunopathologic features that together constitute a unique bullous disease phenotype. There appear to be at least two immunologically distinct subtypes of bullous SLE characterized by the presence or absence of circulating and/or tissue-bound basement membrane zone autoantibodies that recognize type VII collagen. The two subtypes are not clearly distinguishable except by indirect immunofluorescence and/or direct immunoelectron microscopy. In patients without circulating antibodies, immunoelectron microscopy is required to distinguish between the two subtypes. Patients with autoantibodies to type VII collagen are similar but not identical to patients with epidermolysis bullosa acquisita--another bullous disease associated with autoantibodies to type VII collagen. Autoantibodies to type VII collagen in patients with bullous SLE is only one of several lines of evidence that indicate autoimmunity to that protein and susceptibility to SLE are associated phenomena. In addition, there is emerging evidence for an association between epidermolysis bullous acquisita and SLE. There is also evidence that autoantibodies to type VII collagen are pathogenic in bullous SLE (and epidermolysis bullosa acquisita) and that their production is regulated by the class II major histocompatibility complex DR beta 1 allele, 1501 and possibly other DR beta 1 alleles that share a similar sequence of amino acids in the second hyper-variable region. PMID- 8423390 TI - Evidence that involucrin is a covalently linked constituent of highly purified cultured keratinocyte cornified envelopes. AB - The cornified envelop, the terminal product of keratinocyte differentiation, is composed of a variety of covalently cross-linked proteins that form a rigid three dimensional structure. Our present studies show that preparations of intact envelopes prepared from cultured human keratinocytes contain soluble involucrin, keratin, and filaggrin. Sequential extraction with sodium dodecyl sulfate (SDS) and urea followed by sonication produces envelope fragments that are largely free of soluble proteins, including involucrin. Digestion of these highly purified envelope fragments with cyanogen bromide (CNBr) releases a smear of anti involucrin immunoreactive material (40-180 kDa) with two enriched clusters of bands at approximately 52 and 70 kDa. The 52-kDa band cluster co-migrates with products released by CNBr digestion of purified involucrin. The CNBr-mediated release of discrete (52- and 70-kDa) involucrin-immunoreactive bands suggests that many involucrin molecules may be cross-linked at relatively few glutamyl residues/molecule in envelopes prepared from cultured keratinocytes. Moreover, what appears to be cross-linked involucrin can be localized on highly purified sonicated envelope fragments using colloidal gold electron microscopy. These results provide evidence that involucrin is a cross-linked component of the keratinocyte marginal band. PMID- 8423391 TI - Culturing keratinocytes and fibroblasts in a three-dimensional mesh results in epidermal differentiation and formation of a basal lamina-anchoring zone. AB - The purpose of this study was to characterize an in vitro co-culture model in which fibroblasts grown in a three-dimensional nylon mesh were recombined with human keratinocytes. The cultures were kept for 3 and 5 weeks and then processed for electron microscopy. Keratinocytes showed reconstruction of an epidermis consisting of a basal layer with hemidesmosomes, a stratified epithelium with tonofilaments and desmosomes, a granular layer with keratinosomes and keratohyaline granules, and a transitional stratum corneum. Anchoring filaments, lamina densa, anchoring fibrils, bundles of elastin-associated microfibrils (diameters 10 nm) and fine collagen fibrils were formed. Collagen fibrils near the epidermis were much thinner than those in the lower levels. The present study shows that the dermal model containing metabolically active fibroblasts in their natural environment will support epidermal morphogenesis and differentiation including the formation of a basal lamina and anchoring zone. PMID- 8423392 TI - Mechanisms of UV-induced inflammation. AB - The inflammation produced by exposure to ultraviolet (UV) light has been well documented clinically and histologically. However, the mechanisms by which mediators induce this clinical response remain poorly defined. It is clear that photochemistry occurring after UV absorption must be responsible for initiating these events. Some of these underlying mechanisms have been defined. After exposure to UV light, the formation of prostaglandins and the release of histamine are increased. In addition to an increase in the quantity of these mediators, an increase in sensitivity of irradiated tissue to agonist stimulation also occurs. This increased sensitivity may cause tissue to respond to agonist levels previously present. Phospholipase activity also increases, making more substrate available for prostaglandin formation. Oxygen radical-induced peroxidation of membrane lipids caused by irradiation may contribute to increased phospholipase activity. Oxygen-free radicals also participate in sunburn cell formation and in UV-induced decreases in Langerhans cell numbers. Several enzymatic and non-enzymatic mechanisms are present in skin for reducing these highly reactive oxygen species. PMID- 8423393 TI - Characterization, barrier function, and drug metabolism of an in vitro skin model. AB - We have characterized an in vitro skin model consisting of neonatal keratinocytes and fibroblasts grown on a nylon mesh. To produce a dermal model, fibroblasts were seeded onto nylon mesh and grown for 4 weeks until a physiologic dermal-like matrix was formed. This matrix was found to consist of collagens I and III, fibronectin, and glycosaminoglycans. Keratinocytes were then seeded onto the dermal model and the co-culture was grown at the air/liquid interface. A differentiated epidermis with distinct basal, spinous, granular, and stratum corneum layers was formed. When incubated in the presence of keratinocytes, fibronectin immunofluorescence increased throughout the dermis compared to cultures incubated similarly in the absence of keratinocytes. A basement membrane zone rich in laminin, collagen IV, and heparan sulfate proteoglycan was detected. The epidermis, isolated from the co-culture by thermolysin digestion, was analyzed for differentiation markers. K1 keratin (67-kDa) and involucrin were detected by immunologic techniques. Ceramide lipids (types III and IV), thought to be important in barrier function, were detected by thin-layer chromatography. The permeability of the co-culture to a panel of compounds, including [3H]-water, was determined using Franz and side-by-side diffusion cells. The permeability coefficient for water was of the same order of magnitude as that determined for neonatal foreskin. The co-culture also showed selective permeability to a panel of compounds of differing lipid solubility. This co-culture metabolized [3H] testosterone to a profile of metabolites similar to that of neonatal foreskin. We believe that this in vitro skin model will be useful for the study of drug permeability and metabolism. PMID- 8423394 TI - Epidermal cytokines in murine lupus. AB - Murine lupus and the analogous human disease systemic lupus erythematosus (SLE) in humans are characterized by multisystem disease accompanied by the production of numerous serum autoantibodies. The classic model of murine lupus is the New Zealand black mouse (NZB). In this strain anti-DNA antibodies are the most specific marker for the presence of murine lupus, in that this autoantibody parallels both the development and activity of the disease. Exposure to ultraviolet (UV) radiation is known to exacerbate the disease in both the murine and the human disease. UV irradiation of the skin increases serum levels of certain cytokines including interleukin-1 (IL-1), IL-6, and granulocyte/macrophage-colony stimulating factor (GM-CSF), which can influence B- and T-cell function. Recent studies have focused on the role of cytokines in SLE. We hypothesize that the ultraviolet (UV)-induced exacerbation in NZB mice in part is mediated by UV-induced cytokines such as IL-1. Eight-week-old female NZB and DBA/2 mice were exposed to UV irradiation. Sera and supernatants from spleen cell cultures were assayed for anti-DNA antibodies. After UV exposure, NZB mice showed a marked increase in such antibodies. Skin from both strains of mice was probed for IL-1 alpha mRNA before and after UV irradiation. At 24 h, DBA/2 mice had a slight increase in mRNA coding for IL-1 alpha, whereas a much greater increase in skin IL-1 alpha was seen in the NZB skin. This increase in IL-1 mRNA was associated with similar increases in IL-1 bioactivity. These data suggest that the mechanism underlying the UV-induced exacerbation of lupus is mediated in part by the cutaneous production of IL-1. PMID- 8423395 TI - Skin organ culture model for examining epidermal melanization. AB - To clarify the regulatory mechanism of skin pigmentation through epidermal proliferation and differentiation, organ cultures of pigmented guinea pig skins have been studied for their melanogenic responses to exogenous stimuli. Melanogenic activity was measured by both tyrosinase activity assessed by observing release of 3H2O from tissue after incubation with 3H-tyrosine and melanin synthesis, indicated by the incorporation of 14C-2-thiouracil into tissue. Those skin explants that were maintained in serum-free media under air conditions equilibrated with 5% CO2, 50% O2, and 45% N2 formed new, chemically analyzable pigment in the tissue within 4 d of culture. This melanization was accompanied by an increased number of melanocytes as well as by enhanced tyrosinase activity and elevated melanin synthesis. This organ culture system responded well to known tyrosinase inhibitors such as phenylthiourea, which decreased melanogenic activity. Of the arachidonic acid metabolites tested, PGE2, LTC4, LTB4, and 5-HETE were found to significantly stimulate melanogenic activity as indicated by tyrosinase activity, whereas PGF2-alpha, 12-HETE, and 15-HETE did not. Among several known growth factors, only bFGF significantly stimulated melanogenesis in the organ cultured melanocytes. TGF-alpha, which increased DNA synthesis, had a slightly stimulating effect on melanogenesis, whereas TGF beta, which inhibited DNA synthesis, did not stimulate melanogenesis, but rather slightly decreased it. 8-methoxypsoralen+ultraviolet A-treated skin that underwent a marked pigmentation within 14 d in vivo demonstrated enhanced melanogenesis in the organ culture system in proportion to its in vivo pigmentation. Our organ culture system might provide an opportunity to examine the mechanism of action of epidermal melanization. PMID- 8423396 TI - Sunlight and skin-associated lymphoid tissues (SALT): if UVB is the trigger and TNF alpha is its mediator, what is the message? AB - The damaging effects on cutaneous immunity of low-dose ultraviolet B (UVB) radiation in sunlight are genetically determined in mice. Polymorphic alleles at the Tnf alpha and Lps loci dictate whether mice are UVB susceptible or resistant, i.e., develop contact hypersensitivity or not when hapten is painted on UVB exposed skin. In mice, UVB susceptibility is mediated almost exclusively by tumor necrosis factor-alpha (TNF alpha). Circumstantial evidence implicates urocanic acid (UCA) in the stratum corneum as the photoreceptor for UVB, and recent results suggest that cis-UCA in turn instigates the intraepidermal accumulation of TNF alpha. It is hypothesized that TNF alpha interrupts the induction of contact hypersensitivity by preventing epidermal Langerhans cells from carrying hapten to the draining lymph node, where activation of naive, hapten-specific T cells must first occur. The phenotypic traits of UVB susceptibility (UVB-S) and UVB resistance (UVB-R) have now been documented in human beings, and the frequency of UVB-S is high (approximately 40-45%) in both Caucasians and individuals with deeply pigmented skin. Because the frequency of UVB-S is extremely high in patients with biopsy-proved basal and squamous cell skin cancer, this trait appears to be a risk factor for this disease. The unexpectedly high frequency of UVB-S in human beings, including black-skinned persons, implies that the trait is not perceived by evolutionary processes as deleterious. The possible selective advantages conferred by alleles that determine UVB-S are discussed with respect to cutaneous infections in which mortality and morbidity are primarily mediated by immunopathogenic processes. PMID- 8423397 TI - Phototesting in lupus erythematosus. AB - Ultraviolet (UV) irradiation is a major factor in the pathogenesis of certain variants of cutaneous lupus erythematosus. Photosensitivity constitutes one of the criteria of the American Rheumatism Association for the diagnosis of systemic lupus erythematosus, which further emphasizes its importance. The pathomechanism of UV-induced lupus erythematosus remains unknown. The characterization of photosensitive subacute cutaneous lupus erythematosus (SCLE) by Gilliam and Sontheimer has led to a new approach. Through the development of standardized test methods it has became possible to reproduce cutaneous lesions in the UV-A and UV-B spectrum. These standardized test methods allow a better definition of photosensitivity than clinical history does. Recent clinical data show that besides SCLE another variant, lupus erythematosus tumidus, also reveals pronounced photosensitivity. In this review article phototest procedures, phototest results, and clinical correlations in different subgroups are discussed. PMID- 8423398 TI - Subcellular distribution of tyrosinase and tyrosinase-related protein-1: implications for melanosomal biogenesis. AB - Are tyrosinase, encoded at the albino locus, and tyrosinase-related protein-1 (TRP-1), encoded at the brown locus, similarly distributed in melanocytes? We determined the subcellular distribution of tyrosinase and TRP-1 using density fractionation of postnuclear supernatants from mouse melanoma cells of defined genotype followed by immunoblotting with specific antipeptide sera. In highly melanized cells, the majority of tyrosinase cosedimented on Percoll density gradients with visible melanin and with the peak of DOPA incorporation, confirming its presence predominantly in stage III-IV melanosomes. In contrast, the distribution of TRP-1 was limited to a less-dense melanosomal compartment, devoid of melanin. In amelanotic or minimally melanized cells, the majority of tyrosinase shifted into these lighter peaks. To explore a suspected relationship between lysosomes and melanosomes, we analyzed the distribution of lysosome associated membrane protein-1 (LAMP-1). An overlap in the distribution of LAMP-1 and TRP-1 was demonstrated by immunomicroscopy and confirmed by immunoisolation. LAMP-1 was not present in the dense, melanin-rich melanosomal peak on gradient analysis. TRP-1 from melanoma cells homozygous for the brown mutation is not fully glycosylated, is more rapidly degraded, and is restricted in its distribution compared to its wild-type counterpart. In these mutant cells, all melanosomal compartments contain LAMP-1. Our results demonstrate that in wild type cells the majority of tyrosinase eventually localizes to stage III-IV melanosomes. TRP-1 is limited to a less dense melanosomal compartment that is also LAMP-1 positive. The existence of this compartment suggests that it may represent a common step in the biogenesis of melanosomes and lysosomes. PMID- 8423399 TI - Human trichohyalin gene is clustered with the genes for other epidermal structural proteins and calcium-binding proteins at chromosomal locus 1q21. AB - Trichohyalin is a major differentiation product of hard keratinizing tissues such as the inner root sheath and medullary cells of the hair follicle and the filiform papillae of the tongue, as well as terminally differentiating epidermal cells. It consists largely of quasi-repeating peptide repeats and functions primarily as an intermediate filament-associated protein in these tissues. By mapping with human-rodent somatic cell hybrids and fluorescent in situ hybridization, we demonstrate that its gene maps to chromosomal region 1q21. Interestingly, genes encoding several other structural proteins expressed during terminal differentiation in the epidermis map to this region, as do also several members of the S-100 class of small calcium-binding proteins. PMID- 8423400 TI - T- and B-cell abnormalities in systemic lupus. AB - A wealth of T- and B-cell abnormalities has been described in systemic lupus erythematosus (SLE). In general, T and B cells manifest evidence of intense in vivo activation, yet perform poorly when tested for in vitro responsiveness to exogenous antigen. The immune system behaves as if its commitment to self responsiveness overrides the normal capacity to respond to immunogenic challenges. Many immunoregulatory abnormalities have been characterized, but their relation to disease remains controversial. Although a widely accepted unifying hypothesis for the vigorous autoantibody production and associated immune deficiency of SLE is lacking, it seems certain that the profound abnormalities of T and B cells in SLE are intertwined with the fundamental cause of this illness. PMID- 8423401 TI - Molecular characterization of the Ro/SS-A autoantigens. AB - Molecular techniques have recently revealed that there are several immunologically distinct Ro/SS-A antigens. Three genes encoding putative Ro/SS-A protein antigens with calculated masses of 46, 52, and 60 kD have been isolated. The encoded amino acid sequence of each is quite dissimilar. The 46-kD antigen is calreticulin (CR), a highly conserved calcium-binding protein that resides predominately in the endoplasmic reticulum where it may be involved in protein assembly. Although CR has recently been confirmed to be a new human rheumatic disease-associated autoantigen, its relationship to the other components of the Ro/SS-A ribonucleoprotein has become somewhat controversial owing predominately to the fact that recombinant forms of calreticulin have not displayed the same pattern of autoantibody reactivity possessed by the native form of this protein. The 52-kD antigen most likely resides in the nucleus and may be involved in the regulation of gene expression. The cellular location and function of the 60-kD antigen is uncertain but studies indicate that it is a RNA-binding protein. The 46- and 60-kD antigens share homology with foreign polypeptides, suggesting that an immune response initially directed against a foreign protein may give rise to the autoimmune response directed at cross-reacting self proteins. PMID- 8423402 TI - Dendritic epidermal T cells: lessons from mice for humans. AB - Dendritic epidermal T cells (DETC) in mice form part of a primitive system of epithelial-resident T cells characterized by the expression of gamma delta T-cell receptors (TCR). Critical attributes that characterize DETC include their highly restricted T-cell receptor gene utilization, proliferation and maturation within epidermis, a capacity to kill relevant skin-derived tumor targets, and the ability to modulate immune responses that are initiated and expressed in skin. Contemporary knowledge suggests that DETC and the related skin-directed gamma delta T cells found in humans play important roles in maintaining the immunologic integrity of skin. PMID- 8423403 TI - High-affinity fatty acid-binding activity in epidermis and cultured keratinocytes is attributable to high-molecular-weight and not low-molecular-weight fatty acid binding proteins. AB - Fatty acid-binding proteins (FABPs) are abundant low-molecular-weight cytosolic proteins in tissues involved in fatty acid (FA) metabolism. Because epidermis is also an active lipogenic tissue, we examined cytosols from murine and porcine epidermis and cultured human keratinocytes and fibroblasts for FABPs. High affinity FA-binding activity was present in both epidermis and differentiated keratinocytes, whereas no high-affinity FA-binding activity was found in cultured human fibroblasts or undifferentiated keratinocytes. By column chromatography, a single binding peak was identified in the high (90-100 kDa)-molecular-weight range and no binding activity was evident in the low (14-15 kDa)-molecular-weight range, where conventional FABPs elute. Moreover, rabbit anti-rat heart FABP, anti rat intestine FABP, and anti-rat liver FABP antisera did not identify proteins in the 14-15-kDa range in murine epidermal cytosol by Western immunoblots, whereas the anti-rat-heart antibody recognized a protein of approximately 32 kDa. Isoelectric focusing of differentiated keratinocyte cytosol demonstrated a single FA-binding peak having a pI of approximately 4.0. Analysis of this binding peak by SDS-PAGE revealed peptides of approximately 66 and 38 kDa. These findings suggest the possibility that the FA-binding protein in keratinocyte cytosol normally exists as a heterodimer. Western immunoblots of both differentiated keratinocyte cytosol and keratinocyte-conditional media stained with a rabbit anti-human serum albumin antibody identified a protein of approximately 67 kDa, but the electrofocused fraction did not react with this antibody. Thus, epidermis and differentiated keratinocytes possess high-affinity cytosolic FA-binding activity that cannot be ascribed either to conventional low-molecular-weight FABPs or to albumin. PMID- 8423404 TI - Accumulation of gamma delta T cells in chronic cutaneous lupus erythematosus. AB - Monoclonal antibodies (moAbs) that recognize common or variable determinants of the gamma delta T-cell receptor (TcR) were used to assess gamma delta T-cell distribution on biopsy specimens and/or peripheral blood leukocytes (PBL) from 30 patients suffering from chronic cutaneous lupus erythematosus (CCLE). CD3+/gamma delta TcR + T cells were evaluated in 15 biopsies from patients with CCLE lesions, their numbers varying from 0.5 to 15.0% of all intralesional CD3 +T cells present. In all specimens from lesional skin gamma delta TCR+T cells were BB3 + and/or Ti gamma A +, indicating predominant use of the V gamma 2/V delta 2 phenotype. In the CCLE lesions the intraepidermal V gamma 2/V delta +T cells were observed in close vicinity to the damaged basal keratinocyte (KC) layer, and also randomly scattered among the densely packed inflammatory infiltrate in the dermis. In contrast to the immunohistologic findings, no numerical increase of gamma delta TcR+T cells could be observed among PBL from 28 of 30 CCLE patients. Only one CCLE patient being treated with hydroxychloroquine for two months had 15% CD3 +/gamma delta TcR+T cells among the PBL. Based on the immunohistologic findings one may infer that in CCLE, a skin-restricted form of LE, V gamma 2/V delta 2 +T cells expand extrathymically to an as yet unknown stimulus. One may also propose that these gamma delta T cells--based on their cytotoxic capacity- may contribute to the epidermal damage. It remains to be determined whether the extrathymic expansion of V gamma 2/V delta 2 +Tells occurs within lesional skin or in the periphery within subsequent recruitment into skin lesions. The results obtained by fluorescence-activated cell sorter analysis favor the first possibility. PMID- 8423405 TI - Antibodies to mycobacterial 65-kDa heat shock protein and other immunodominant antigens in patients with psoriasis. AB - An association of microbial agents and autoimmunity has been suggested for the pathogenesis of psoriasis. Mycobacteria are common environmental microbes and their antigens, especially the highly conserved mycobacterial 65-kDa heat shock protein (hps65), have been implicated in the pathogenesis of autoimmune arthritis and other idiopathic diseases. In this context, we investigated a possible mycobacterium-induced humoral immune response in psoriasis. Sera from 17 patients with chronic plaque-type psoriasis were studied by immunoblotting using the whole sonicate of Mycobacterium tuberculosis and purified recombinant mycobacterial hsp65. Immunoblot analysis demonstrated that 58% of the psoriasis patients compared to patients with acne and DLE, and normal controls showed strong antibody activity to 65-kDa and 48/45 doublet antigens from M. tuberculosis sonicate, whereas 47% of the patients showed antibody activity to mycobacterial hsp65. Only 10-20% of the patients had an antibody response to 16-kDa and 80-kDa antigens. Similar antibody activity to 65 kDa and 48/45 kDa was also found consistently with eight different sonicated mycobacterial species by immunoblotting, indicating that these seroreactive antigens are crossreactive and are present in common environmental mycobacteria. Antibody activities to both mycobacterial 65-kDa and hsp65 showed a positive correlation (r = 0.76) with the psoriasis disease activity, whereas antibodies to 48/45-kDa doublet antigens showed a weak correlation (r = 0.54). By enzyme-linked immunosorbent assay (ELISA), 47% of the psoriasis patients showed significantly elevated antibody titers to hsp65 (p < 0.003) as compared to control groups, and the antibody response by ELISA also showed a significant positive correlation (r = 0.76) with disease activity. Anti-mycobacterial antibody activity may be related to severity of disease and may be useful in monitoring disease activity in psoriasis. PMID- 8423406 TI - Neonatal lupus erythematosus. AB - Neonatal lupus erythematosus (NLE) is an autoimmune disease whose major findings are subacute cutaneous lupus erythematosus (SCLE) skin lesions and congenital heart block. Babies have maternal anti-Ro/SSA, anti-La/SSB, or anti-U1RNP autoantibodies. Anti-Ro/SSA are the predominant autoantibodies, having been found in about 95% of cases. The autoantibodies pass through the placenta from mother to child. Skin disease resolves at about the time that maternal autoantibodies can no longer be detected in the baby. NLE therefore provides the strongest clinical evidence that autoantibodies are involved in at least some manifestations of lupus erythematosus, but there is as yet no definitive evidence implicating autoantibodies in the disease process. Skin disease usually begins after birth, is transient, and does not result in scarring. Cardiac disease begins in utero, and the heart block is almost always permanent. Many babies require pacemakers, and about 10% die from complications related to cardiac disease. In some cases, transient liver disease or thrombocytopenia have been observed. Individuals who had NLE usually have healthy childhoods but may develop autoimmune disease in adulthood. Whether the later development of autoimmune disease is a common or an unusual event is not yet known. Mothers of babies with NLE may be asymptomatic initially, but with time usually develop symptoms of autoimmune disease. The most typical constellation of symptoms in our group of approximately 30 mothers of babies with NLE is that of Sjogren's syndrome. Most babies exposed to anti-Ro/SSA autoantibodies during gestation will not develop NLE. There is no test to determine prospectively which babies will be affected. Treatment during gestation is still controversial and, if attempted, should be reserved for fetuses with potentially life-threatening disease. Treatment after birth consists of topical management for skin disease and pacemaker implantation, if necessary, for heart block. Systemic steroids may be given for serious internal disease. PMID- 8423407 TI - Control of fibrosis in systemic scleroderma. AB - Scleroderma is characterized by an excessive deposition of collagen in all involved organs. This is due to an overproduction of extracellular matrix (ECM) molecules following induction of gene expression, whereas there is no evidence that the composition of the connective tissue matrix is altered. Several in vivo studies and in vitro experiments suggest that a close interaction between inflammatory cells and fibroblasts is required for the initial activation of fibroblasts. TGF-beta presumably plays an important role, but other cytokines, e.g., PDGF or FGF, may also be involved. Many of the ECM molecules have been shown to interact closely with fibroblasts and provide signals that regulate fibroblast metabolism. The cellular response towards those signals is a further aspect of fibrosis that has attracted attention during recent years. The altered expression of receptor proteins on the cell surface of scleroderma fibroblasts for example might explain in part the lack of down-regulation of collagen synthesis in late phases of the disease. This review summarizes the alterations of connective tissue in scleroderma, and discusses the role of cytokines as well as the ECM for the regulation of fibroblast function and their implication for the development of fibrosis. PMID- 8423408 TI - Normal molecular weight of type VII collagen produced by recessive dystrophic epidermolysis bullosa keratinocytes. AB - Studies of the recessive dystrophic form of epidermolysis bullosa (RDEB) have suggested that an abnormality in type VII collagen may be involved in the pathogenesis of this disorder. Indirect immunofluorescence studies have shown that the staining for type VII collagen along the dermal-epidermal junction is markedly reduced or absent in all but rare cases of severe, generalized RDEB. These findings imply that the genetic defect may involve type VII collagen but do not exclude the possibility that the alterations demonstrated are secondary, for example, to nonspecific proteolysis of type VII collagen. To evaluate the ability of cells of affected patients to produce type VII collagen, we cultured keratinocytes from a severely affected patient and immunoprecipitated type VII collagen from the cells. Keratinocytes were metabolically labelled with 35S methionine, and solubilized cell extracts were reacted with antibody to type VII collagen. The results indicate that the patient's keratinocytes synthesize type VII collagen and that the M(r) of the protein synthesized does not differ from that of an unaffected control. Because cultured cells from a patient severely affected with recessive dystrophic epidermolysis bullosa produce type VII collagen, the genetic defect, at least in this patient, is unlikely to reside in a major truncation of the type VII collagen molecule. PMID- 8423409 TI - L-tryptophan and the eosinophilia-myalgia syndrome: current understanding of the etiology and pathogenesis. AB - The eosinophilia-myalgia syndrome (EMS) is a newly recognized illness that occurred in an epidemic form during the summer of 1989. The illness was characterized in the acute phase by myalgia and eosinophilia, followed in many patients by chronic cutaneous lesions, progressive neuropathy, and myopathy. EMS was associated with ingestion of L-tryptophan, an essential amino acid marketed as a nutritional supplement but widely used as a therapeutic agent. Evidence of abnormal L-tryptophan metabolism has been described in patients with EMS, and most likely reflects increased activity of indoleamine 2,3-dioxygenase, the rate limiting enzyme of tryptophan metabolism. A contaminant identified in EMS associated L-tryptophan preparations has been isolated and characterized, but its biologic effects and role as the etiologic agent in EMS remain to be established. Pathologic observations and experimental studies indicate that eosinophils, mononuclear inflammatory cells, and fibroblasts are potential effector cells, and interleukin-5 and transforming growth factor-beta are important mediators in the pathogenesis of the syndrome. Although few new cases of EMS occurred following the withdrawal of L-tryptophan, affected patients continue to manifest late sequelae of the disease, including dermal fibrotic conditions. This tragic outbreak of a newly recognized illness has focused interest on the role of chemical and environmental agents in the pathogenesis of various idiopathic illness characterized by tissue inflammation and fibrosis. PMID- 8423410 TI - Pheochromocytoma with multiple system failure. AB - Pheochromocytomas may present with a variety of clinical manifestations. A rare presentation is that of multiple system failure, which may be fatal if not promptly diagnosed and treated. The case of a patient who developed multiple system failure from a pheochromocytoma is presented. PMID- 8423411 TI - Primary non-Hodgkin's lymphoma of the central nervous system. AB - In the past few years, non-Hodgkin's lymphoma of the central nervous system has been the object of increasing attention because of the recent rise in its incidence. Part of this increase can clearly be attributed to the AIDS epidemic. This tumor responds unsatisfactorily to the traditional treatments. Various studies to determine the most effective treatment modality have yielded inconclusive results. A concise review of non-Hodgkin's lymphoma of the central nervous system is presented along with a case study of cerebellar non-Hodgkin's lymphoma. PMID- 8423412 TI - Jet-ski injury: a case history. AB - A case history is presented which involves a patient who sustained severe vaginal lacerations as a result of falling backwards off of a jet-ski. The surgical repair of the lacerations and postoperative course is discussed. An extensive search of the literature has failed to reveal any published reports of a similar injury. PMID- 8423413 TI - Effective communication: a powerful risk management tool. AB - Physicians can employ communication techniques to improve patient diagnoses, outcomes, and satisfaction and ultimately to decrease their risk of malpractice suit. The skills outlined in this article form the basis of the Miles Program for Physician-Patient Communication of which the author is a participant. PMID- 8423414 TI - ECG of the month. Euthanasia and abortion. PMID- 8423415 TI - Two-way street. Ventricular ectopic activity. PMID- 8423416 TI - Velopharyngeal insufficiency. AB - Velopharyngeal insufficiency (VPI) is a relatively uncommon diagnosis and requires an understanding not only by the otolaryngologist but also by primary care physicians who are often the first to recognize its presence. An appreciation of the speech pathology that occurs in VPI assists in the often challenging diagnosis. Although VPI is more commonly seen in the pediatric population, there are multiple causes for its presence in adults and children. We review the anatomy of the velopharynx and the etiology of VPI, as well as the evaluation and the management of these patients. PMID- 8423417 TI - The relationship between the St. Thomas and Oswestry disability scores and the severity of low back pain. AB - OBJECTIVE: To investigate the relationship between the two disability questionnaires and low back pain severity using the visual analog scale (VAS). DESIGN: One hundred sixty-two chiropractic patients with low back pain were interviewed using an interactive microcomputer system. The sources of data were the St. Thomas and Oswestry disability questionnaires for disability and a VAS for pain severity. All were inherent in the computer interview system. SETTING: The computer review system was set up in the Anglo-European College of Chiropractic outpatient clinic. PATIENTS, PARTICIPANTS: One hundred sixty-two consenting new patients from the general community suffering from low back pain who attended the Anglo-European College of Chiropractic outpatient clinic were asked to participate. MAIN OUTCOME MEASURES: Main outcome measures were the St. Thomas and Oswestry disability questionnaires for disability and the VAS for low back pain severity. RESULTS: There was a moderate correlation between the scores from the St. Thomas and Oswestry disability questionnaires (r = .77, p < .0001). A low but significant correlation score was found when comparing the St. Thomas and Oswestry disability score with pain severity (0.38, p < .0001, and 0.47, p < .0001, respectively). CONCLUSION: The consequence of pain (disability) appears to have a weak relationship to pain severity. Despite the moderate correlation between the two disability questionnaires, it is suggested that they are not interchangeable. PMID- 8423418 TI - Health psychology: implications for chiropractic research. AB - There is a body of research in health psychology which is of potential interest to chiropractic researchers. Health psychology is an area of psychology devoted to the maintenance of health and prevention of illness from a biopsychosocial rather than a biomedical perspective. Many of the research topics found in the psychological literature, and the model of nervous system involvement in somatic illness, are similar to the research interests and models that can be found in the chiropractic literature. There may be benefits to chiropractic research in collaboration with established researchers in psychology, many of whom are funded by substantial federal grants. PMID- 8423419 TI - Patient satisfaction with chiropractic care. AB - OBJECTIVE: To determine the attitudes of patients regarding the process and result of chiropractic care and to identify patient characteristics which might predict satisfaction. DESIGN: Mailed survey consisting of a patient satisfaction questionnaire composed of 32 attitude statements accompanied by a five-point Likert scale, a personal information questionnaire and a doctor questionnaire providing clinical information about the patient. SETTING: Nonrandom sample of chiropractic doctors engaged in private practice. PATIENTS: Survey questionnaires were mailed to 541 new and returning chiropractic patients seeking care between June 1988 and August 1989, with a response rate of 69.5% (n = 376). RESULTS: Survey items were organized into scales and subscales from which response means were calculated reflecting attitudes about specific dimensions of care. Women responding to the survey were slightly more satisfied with the care they received than men, but other patient characteristics did not influence response means for questions referring to general satisfaction. Patients were most satisfied with the accessibility of their doctors and least satisfied with the financial aspects of treatment--especially those who reported lower incomes and no insurance coverage. Finally, among a variety of factors which might influence patient satisfaction, we found that the patient's perception of treatment outcome was the most important predictive variable. A slightly greater degree of dissatisfaction was reported by a small percentage (12%) of patients who also reported that there was either no improvement in their health problem, or minimal improvement, following chiropractic care. CONCLUSIONS: Patients expressed high levels of satisfaction with their doctors and the care they received. Although women were slightly more satisfied than men, other patient characteristics such as level of education, income, employment status or previous chiropractic care did not influence response means. Future research is needed to determine if the way in which chiropractic care is rendered affects patient satisfaction. PMID- 8423420 TI - Manual force, mechanically assisted articular chiropractic technique using long and/or short level contacts. AB - OBJECTIVE: To identify what has been theorized and/or written about the manual techniques generically classified as manual force, mechanically assisted articular chiropractic technique that utilize either short-lever contacts or a combination of short- and long-lever contacts. DATA SOURCES: A search of the Medline bibliographic database using MeSH key words (chiropractic/methods; osteopathic manipulation) was performed. The Index to Chiropractic Literature and Chiropractic Research Abstracts Collection (CRAC) were searched for the past 10 yr using the key terms of chiropractic-methods; chiropractic techniques; manipulation, chiropractic, manipulation, osteopathic, manipulation, spinal; and manipulation, joint. A hand search of textbooks was undertaken as well as review of the references included in books, monographs and collected papers. STUDY SELECTION: Studies in English were included, but it must be noted that these include publications that are not peer reviewed or refereed in any way. DATA EXTRACTION: Descriptions that discussed aspects of manipulative procedures with the characteristics were extracted by a single author. DATA SYNTHESIS: Very little was found or accessible with traditional methods of literature retrieval. Descriptions of characteristics for the attributes of techniques which use mechanical assistance to impart a force applied to specific contacts in combination with short and long levers are provided. The technique procedures which fall into this category use the mechanical assistance of specialized table parts, including drop sections and distracting sections, to achieve functional changes in the vertebral three-joint complex. Conjecture and speculation as to the advantages of mechanical assistance are presented. CONCLUSIONS: This type of review is considered the first step in the evaluative process for a chiropractic technique procedure. It demonstrates that very little has been written in an accessible fashion relative to techniques which are classified as manual force, mechanically assisted articular chiropractic techniques that utilize either short lever contacts or a combination of short- and long-lever contacts. Controlled prospective clinical trials to evaluate efficacy in using these technique procedures are nonexistent. Furthermore, there are no comparison studies to determine whether the techniques which fit into this category are any more effective or efficient in producing a positive clinical outcome than techniques in other categories. The proponents of these technique procedures should be performing the studies and publishing the results that either support or deny the usefulness of these procedures. PMID- 8423421 TI - Idiopathic osteonecrosis of the hip. AB - This article discusses the case of a 77-yr-old man suffering from osteonecrosis of the femur head. Diagnostic imaging and clinical manifestations are discussed along with the appropriate patient management. If untreated, osteonecrosis of the hip will often result in severe degenerative joint disease. Successful treatment depends on early diagnosis. PMID- 8423422 TI - Chiropractic magazines. PMID- 8423423 TI - Osteomyelitis of the femur head in a pediatric patient. AB - A case of posttraumatic osteomyelitis of the femur head is discussed. This article emphasizes the importance of radiographic examination in posttraumatic cases. We describe the common radiographic appearance of osteomyelitis, and then the appropriate mode of treatment for the patient via antibiotic therapy and surgical debridement/curettage. PMID- 8423424 TI - The precipitation or aggravation of musculoskeletal pain in patients receiving spinal manipulative therapy. AB - Complications of chiropractic adjustments or manipulations can be considered either benign or catastrophic. Although there are a number of papers on the more severe complications, the precipitation of benign and often reversible symptoms following chiropractic care has been largely ignored in the literature. This article discusses six cases where pain symptomatology was either aggravated or precipitated at a location other than the presenting complaints. Two potential causes of these complications are discussed: a) the use of long levers, which can exert force on nonsymptomatic joints in the extremities, and b) the manipulation of biomechanically dysfunctional but asymptomatic joints. PMID- 8423425 TI - The physics of spinal manipulation: work-energy and impulse-momentum principles. AB - The purpose of this article was to illustrate that the work-energy and impulse momentum relations are derived from Newton's second law, and, thus, will give identical results to F = ma for any force system analysis. It should be noted that using the work-energy or the impulse-momentum approach requires that all external forces acting on a system are considered. Furthermore, it must be emphasized that any force acting internal to the system (e.g., muscular forces in the system "chiropractor") do not enter these equations. PMID- 8423427 TI - Commentary: shades of straight--diversity among the purists. PMID- 8423426 TI - Mechanical force, manually assisted short lever chiropractic adjustment. PMID- 8423428 TI - Forefoot pain associated with muscle strain in the lower extremity. PMID- 8423429 TI - The Hmax/Mmax ratio as an outcome measure for acute low back pain. AB - OBJECTIVE: To evaluate the use of the Hmax/Mmax (H/M) ratio as an outcome measure for acute low back pain and to determine the change of this ratio in acute low back pain patients treated with spinal manipulation. DESIGN: Randomized clinical trial. SETTING: Chiropractic college teaching clinic. PATIENTS: Thirty-six patients with acute low back pain (pain of less than 2 wk duration) were referred by clinicians of the teaching clinic. Eligibility criteria for inclusion into the study consisted of the following: a score of eight or more on the Oswestry questionnaire, 33 mm or greater on a 100-mm visual analog scale, no involvement in litigation related to the low back pain complaint, patient not pregnant and no physical or electrodiagnostic signs of nerve root entrapment. INTERVENTIONS: The patients were randomly assigned to either a treatment or control group. The treatment group (n = 17) received treatment deemed appropriate by the clinician as long as it included a side-lying manipulation to the appropriate level. The control group (n = 19) received detuned ultrasound, application of a cold pack and 15-30 sec of very gentle soft tissue massage. Patients were treated three to five times over a period of 10 days and were subsequently reevaluated. MEASUREMENTS: The Hmax/Mmax ratio was calculated from the results of electrodiagnostic testing of the posterior tibial nerve. Extension/flexion ratio of the trunk musculature, Oswestry score and Visual Analog Scale score were also measured. MAIN RESULTS: The mean difference between H/M ratios pre- and postintervention for the group treated by chiropractic methods was -0.101 on the left and -0.117 on the right. The mean difference for the control group was 0.038 on the left and 0.036 on the right. Although not statistically significant, trends suggest that at the time of final assessment, the group receiving chiropractic care had improved more than the control group. CONCLUSIONS: The H/M ratio was found to be within normal limits in subjects with acute low back pain. The H/M ratio showed greater change in the group which received spinal manipulation, but the change was subtle. The results indicate that the H/M ratio may be of limited value in clinical practice. PMID- 8423430 TI - Dissociative effect of massed repetition on implicit and explicit measures of memory. AB - Subjects saw or heard words presented once, or repeated 4 or 16 times in massed fashion, and then received an implicit or explicit memory test. Massed repetition did not increase priming on word fragment completion beyond that obtained from a single presentation but did enhance performance on various explicit tests (free recall, recognition, question cued recall, and word fragment cued recall) and an implicit general knowledge test. Modality of presentation affected implicit and explicit word fragment cued tests but did not affect performance on any of the other tests. Levels of processing affected performance on implicit and explicit question cued tests. These results are consistent with a transfer appropriate processing account of dissociations among memory measures and imply that massed repetition promotes conceptual processing but does not entail a repetition of perceptual-based processes responsible for priming on word fragment completion. PMID- 8423431 TI - Incidental concept learning, feature frequency, and correlated properties. AB - Four experiments examined sensitivity to feature frequencies and feature correlations as a function of intentional and incidental concept learning. Feature frequencies were encoded equally well across variations in learning strategies, and although classification decisions in both intentional and incidental conditions preserved correlated features, this sensitivity was achieved through different processes. With intentional learning, sensitivity to correlations resulted from explicit rules, whereas incidental encoding preserved correlations through a similarity-based analogical process. In incidental tasks that promoted exemplar storage, classification decisions were mediated by similarity to retrieved examples, and correlated features were indirectly preserved in this process. The results are discussed in terms of the diversity of encoding processes and representations that can occur with incidental category learning. PMID- 8423432 TI - Phonetic recoding of phonologically ambiguous printed words. AB - Speech detection and matching simultaneously presented printed and spoken words were used to examine phonologic and phonetic processing of Hebrew heterophonic homographs. Subjects detected a correspondence between an ambiguous letter string and the amplitude envelopes of both dominant and subordinate phonological alternatives. Similar effects were obtained when the homographs were phonologically disambiguated by adding vowel marks. The matching of the unpointed printed forms of heterophonic homographs to the dominant and subordinate spoken alternatives presented auditorily was as fast as matching the pointed unambiguous forms to the respective spoken words. This outcome was not obtained when print and speech were not presented simultaneously. These results suggest that printed heterophonic homographs activate the two spoken alternatives they represent and provide further confirmation for fast phonetic recoding in reading. PMID- 8423433 TI - Monitoring changes in cognitive load during reading: an event-related brain potential and reaction time analysis. AB - Factors that contribute to cognitive load during reading were examined using a secondary task procedure. In three experiments, subjects read sets of passages twice in succession while auditory probes were presented. The N1-P2 and P300 components of the event-related brain potential and reaction time (RT) responses to secondary auditory probes were used as measures of load. N1-P2 responses indicated decreased load during the second reading, whereas P300 and RT responses indicated increased load during the second reading. The results are interpreted as reflecting changes in task demands. Lower level elements of the task, such as word recognition and local aspects of comprehension, required fewer resources during the second reading. The N1-P2 reflected this reduction in resource demands. By contrast, the amount of resources devoted to higher level processes, such as comparing the text with one's prior representation and updating memory, increased during the second reading. This resulted from task demands, which emphasized memory of the material. P300 and RT reflected this increase in higher level demands. Results are described in terms of attentional and task demands and are taken as support for a componential description of reading and task difficulty. PMID- 8423434 TI - Accessibility of information about goals during the processing of narrative texts. AB - We used a probe procedure to show that a goal established earlier in a text is active in memory at the point of its achievement. An initial experiment demonstrated that a goal category (began an investigation to nab the THIEF) is accessible, relative to a control condition, following the processing of a goal achievement sentence (had the PURSER brought to his office). The remaining experiments provided evidence against several explanations of this result: (a) that the goal category's accessibility is due to an advantage in the strength of its initial encoding; (b) that the goal category is maintained in memory from the point at which the goal is established; or (c) that the goal category is reinstated at the point of goal achievement as the result of a high-level inference. The results suggest that the goal category is reinstated as the result of a low-level inference similar to the type that links an anaphor and its antecedent. PMID- 8423435 TI - Auditory word recognition: extrinsic and intrinsic effects of word frequency. AB - Two experiments investigated the influence of word frequency in a phoneme identification task. Speech voicing continua were constructed so that one endpoint was a high-frequency word and the other endpoint was a low-frequency word (e.g., best-pest). Experiment 1 demonstrated that ambiguous tokens were labeled such that a high-frequency word was formed (intrinsic frequency effect). Experiment 2 manipulated the frequency composition of the list (extrinsic frequency effect). A high-frequency list bias produced an exaggerated influence of frequency; a low-frequency list bias showed a reverse frequency effect. Reaction time effects were discussed in terms of activation and postaccess decision models of frequency coding. The results support a late use of frequency in auditory word recognition. PMID- 8423436 TI - Mental model organization. AB - Previous research (Radvansky & Zacks, 1991) has shown that the fan effect is mediated not by the number of nominal associations paired with a concept but by the number of mental models into which related concepts are organized. Specifically, newly learned "facts" about different objects in one location are integrated into a single mental model and no fan effect is produced, whereas facts about one object in different locations are not integrated and a fan effect is produced. In 6 experiments we investigated several factors' influence on location-based organization preferences. We found no impact of either article type (definite or indefinite) or object transportability. However, animate sentence subjects (people) reduced preference for location-based organizations. A clear person-based organization emerged by using locations that typically contain only a single person (e.g., phone booth) to make location-based situations less plausible. PMID- 8423437 TI - Renal clearance measurements of electrolytes in embryonic chickens. AB - Plasma ion concentrations and renal functions were examined in chick embryos after 9, 12, and 15 days of incubation. Plasma calcium and phosphate values rise steadily and significantly between days 9 and 15 of development. Values of other plasma ions (sodium, potassium, magnesium, sulfate) remain more or less the same during this developmental period. Glomerular filtration rates and urine flow rates decline steadily and significantly during this period of embryogenesis when expressed relative to body weight. Renal clearance values of calcium decrease between day 9 (16% of filtered calcium is excreted) and day 15 (4%). The latter value is the same as in birds after hatching and indicates that calcium is being conserved by the kidney of the chick during late embryogenesis. Other ions that appear to be conserved by the embryonic kidney are sodium and sulfate, whereas high percentages of filtered phosphate, potassium, and magnesium are excreted. Except for calcium, there was no significant difference between the renal handling of ions by the mesonephric kidney (functional in 9 day embryos) and the metanephric kidney (predominant in the 15 days embryos). PMID- 8423438 TI - Retinoic acid, cortisone, or thyroxine suppresses the mutant phenotype of the eyelid development mutation, lgMl, in mice. AB - The development of the mammalian eyelid is one of the last major morphogenetic events during prenatal development. The eyelids originate as outgrowths of epithelium and mesenchyme above and below the eye; they grow and flatten across the eyes, and meet and fuse firmly but temporarily until several days after birth in mice. In mouse lidgap mutants, eyelid development fails. Previously, we have shown that normal eyelid development can be induced in lidgap mutants by maternal treatment with cortisone or thyroxine. These hormones have been reported to interact with members of a receptor superfamily, and their response elements have been shown to induce transcription of hormone-regulated genes; retinoic acid also interacts with members of this superfamily. The purposes of our study were to examine the joint effect of thyroxine and cortisone on induced eyelid development in lidgap-Miller mutant fetuses (lgMl/lgMl), to test for synergism, and also to test whether retinoic acid also induces eyelid development in this mutant. We found that the combined effect of thyroxine and cortisone was significantly greater than that of either hormone alone, but that the dose-response slopes (log dose, probit scale) for eyelid development induced by cortisone alone or by cortisone preceded by 0.1 mg thyroxine were parallel, suggesting that their action is additive, not synergistic. We found that retinoic acid administered on day 14 of gestation induced eyelid development in lgMl/lgMl mutant fetuses, with a significant dose response and ED50 of 24 +/- 6 mg/kg. The response was treatment-time dependent, with optimal responses after day 14 or 11 treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423439 TI - Developmental anomalies of Xenopus embryos following microinjection of SPARC antibodies. AB - The function of SPARC (Secreted Protein, Acidic, Rich in Cysteine) in early embryonic development was assayed by microinjecting affinity-purified antibodies directed against SPARC into the blastocoel cavity of Xenopus embryos. Microinjection of SPARC antibodies did not appear to interfere with development until late neurulation. By hatching, a broad spectrum of external developmental anomalies were observable, including bent embryonic axes, accentuated ventral masses, shortened embryonic axes, and lack of visible eye pigment. Histological sections of injected embryos demonstrated that lack of visible eye pigmentation was often associated with deformities in eye development. Bending and shortening of the embryonic axis was associated with highly disorganized myotome patterns and loss of segmental boundaries. The results indicate a requirement for SPARC in the early morphological development of several tissues in Xenopus. PMID- 8423440 TI - Relationship between local cell division and cell displacement during regeneration of embryonic Xenopus eye fragments. AB - We examined the relationship between early healing modes and extra cell division (via tritiated thymidine incorporation) during embryonic retinal regeneration. Nasal (N) and dorsal (D) one-third sized eye fragments were surgically created in stage 32 Xenopus laevis embryos. Embryos were injected with tritiated thymidine two days postsurgery (stage 43), and then fixed and processed for autoradiography one day postinjection (stage 46). Histological analysis revealed that all nasal one-third sized fragments showed cell displacement in healing regions. These displaced cells were located in the ventral retinal region and showed heavy thymidine label incorporation. Alternatively, most dorsal one-third sized fragments showed little cell displacement during healing; in addition, no extra thymidine incorporation was evident. A minority of dorsal one-third sized fragments showed cell displacement during healing, and also showed significant local label ventrally through all regions of the eye. Therefore, in both dorsal and nasal one-third sized retinal fragments, when cell displacements were observed during early healing, associated mitosis was apparent in ventral retinal regions. Furthermore, by 60 hours postsurgery, all eyes which showed cell displacements during healing were greater in volume than those eyes which showed little cell displacement. Increases in volume appear to be derived from a combination of both cells migrating in from underlying optic stalk tissue and from related extra cell division during healing. These data further support a model which predicts that specific healing interactions which involve cell displacement during embryonic retinal regeneration and subsequent intercalary growth underlie visuotectal pattern formation. PMID- 8423441 TI - Electroporation of inositol 1,4,5-triphosphate induces repetitive calcium oscillations in murine oocytes. AB - The purpose of these experiments was to determine the effect of electroporation of IP3 into the cytosol of murine secondary oocytes and evaluate any alterations in [Ca2+]i resulting from Ca2+ release from intracellular stores. In addition, we evaluated the effect of ethanol (ETOH) on the release of Ca2+ from intracellular stores. Oocytes were loaded with the Ca2+ indicator fluo-3 by incubation in 100 microliters drops of medium containing 2 microM fluo-3/AM for 60 min at 37 degrees C. Changes in fluorescence were monitored by use of an inverted microscope which had been connected to a spectrofluorometer. Fluorescent intensity measurements were acquired for a minimum of 416 sec time span or up to 1,248 sec, with integration readings of 1 sec duration obtained every 2 sec throughout the measurement period. The experimental design consisted of comparing the rise in [Ca2+]i of fluo-3 loaded secondary oocytes subjected to electroporation in PBS and Ca(2+)-free PBS, each containing 25 microM IP3, to that elicited by PBS and Ca(2+)-free PBS containing a final concentration of 7% ETOH. Non-pulsed control secondary oocytes were placed in PBS + 25 microM IP3 during monitoring of [Ca2+]i fluorescence. Pulsed control secondary oocytes were placed in Ca(2+)-free PBS, subjected to electroporation pulse, and monitored for [Ca2+]i fluorescence. Electroporation of IP3 was accomplished by placing fluo-3 loaded secondary oocytes between the electrodes of a glass slide fusion chamber which had been overlaid with 300 microliters of PBS + 25 microM IP3 or Ca(2+) free PBS + 25 microM IP3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423442 TI - Differences in chromatin condensation during spermiogenesis in two species of fish with distinct protamines. AB - The sperm cells of Mullus surmuletus (family Mullidae, order Perciformes) and Dicentrarchus labrax (family Percichthyidae, order Perciformes) belong respectively to "type I" and "type II" spermiogenesis categorized by Mattei ('70). The protein content in their sperm nuclei consists of two histone-like proteins (Mullus surmuletus) and one typical protamine (D. labrax). In order to correlate the molecular characteristics of these proteins with their function, we have analyzed the molecules in detail and studied at the ultrastructural level the condensation of chromatin during the spermiogenesis in both species. D. labrax has a true protamine of 34 amino acid residues and its sequence (PR4QASRPVR5TR2STAER5V2R4) contains four arginine clusters. The sperm proteins of M. surmuletus contain 110 and 115 amino acid residues and , by their composition (23-24% Lys, 21-22% Arg, 11-12% Ala), they are similar to protamine-like molecules from sperm of molluscs. During the spermiogenesis of D. Labrax, chromatin condensation progresses from small fibro-granular structures (25 +/- 5 nm in diameter), to larger granules (150 +/- 50 nm diameter). M. surmuletus accumulates 25 +/- 5 nm diameter structures in the basal pole of the nucleus; these structures grow till they reach a diameter of 50 +/- 10 nm and finally go through a process of fusion that changes the condensation of chromatin in sperm nuclei, acquiring a homogeneous aspect. These observations show that during spermiogenesis in the studied types, the last stages of chromatin condensation are dependent on the type of nuclear proteins. PMID- 8423443 TI - The chimeric nature of the genome of pea enation mosaic virus: the independent replication of RNA 2. AB - The genome of pea enation mosaic virus (PEMV) consists of two plus-sense RNAs, both of which are required for mechanical transmission. RNA 1 (5706 nucleotides) has strong sequence similarity with members of the luteovirus group, a similarity that is also manifested in the symptomatology, cytopathology and vector transmission of this virus. RNA 2 (4253 nucleotides) is hypothesized to facilitate systemic invasion and mechanical transmission, attributes that distinguish PEMV from the phloem-limited luteoviruses. Sequence analysis of RNA 2 has demonstrated that PEMV is unique among multicomponent viruses in that it lacks 3'- and 5'-terminal homology between its genomic RNAs. Sequence analysis of RNA 2 has identified an open reading frame encoding a putative product of 65K that contains a series of polymerase-like motifs typical of viral RNA-dependent RNA polymerases. This protein sequence lacks homology with the polymerase encoded on RNA 1 of PEMV, instead being more closely affiliated with the polymerases of viruses related to the carmo- and tombusvirus groups. Inoculation of pea protoplasts with RNA transcripts derived from a full-length cDNA clone of RNA 2 has demonstrated that RNA 2 replicates autonomously in the absence of RNA 1, although comparable inoculation of whole plants failed to establish a systemic infection. There is no evidence that RNA 2 encodes structural proteins, suggesting that encapsidation functions are supplied in trans by RNA 1, comparable to the helper-dependent complexes occurring within the luteovirus group. These data suggest that the PEMV genome can be characterized as a symbiotic association of two taxonomically distinct viral RNAs cooperatively interacting in the establishment of a systemic virus infection. PMID- 8423444 TI - Immunocytochemical characterization of p24, a baculovirus capsid-associated protein. AB - An open reading frame (ORF 1) located upstream of the polyhedron envelope protein gene in the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) genome was cloned in-frame into a trpE bacterial expression vector. The fusion protein produced by this construct was used for the preparation of a monospecific antiserum. Western blot analysis of extracts from OpMNPV-infected Lymantria dispar cells and Autographa californica NPV (AcMNPV)-infected Spodoptera frugiperda cells detected a 24K protein late in infection. This antiserum also reacted with a 24K protein in preparations of budded and polyhedra derived virus from OpMNPV and AcMNPV. The 24K protein was not N-glycosylated. Immunoelectron microscopy confirmed that the OpMNPV p24 is associated with nucleocapsids of budded and polyhedra-derived virions. PMID- 8423445 TI - Antibody response to the M2 protein of influenza A virus expressed in insect cells. AB - A recombinant baculovirus expressing the M2 protein from influenza A/Ann Arbor/6/60 (H2N2) virus (AA60 virus) was constructed. The expressed M2 protein was recognized by a monoclonal antibody specific for the M2 protein and comigrated with the M2 protein from cells infected with AA60 virus on SDS polyacrylamide gels. Immunofluorescence studies indicated that the expressed M2 protein was present on the surface of Spodoptera frugiperda (Sf9) cells infected with the recombinant baculovirus. Immunoassays using the expressed M2 protein were able to detect antibodies to the M2 protein in serum samples from humans and ferrets infected with influenza A viruses. PMID- 8423446 TI - Nucleotide sequence and genome organization of tomato leaf curl geminivirus. AB - The genome of tomato leaf curl virus (TLCV) from Australia was cloned and its complete nucleotide sequence determined. It is a single circular ssDNA of 2766 nucleotides containing the consensus nonanucleotide sequence present in all geminiviruses. It has six open reading frames with an organization resembling that of certain other dicotyledonous plant-infecting monopartite geminiviruses, i.e. tomato yellow leaf curl and beet curly top viruses. The regulatory sequences present indicate a bidirectional mode of transcription. A dimeric TLCV DNA clone was constructed in a binary vector and used to agroinoculate three different host species. Typical virus infections were produced, confirming that the single DNA component is sufficient for infectivity. PMID- 8423447 TI - An analysis of the complete sequence of a sugarcane bacilliform virus genome infectious to banana and rice. AB - The genome of sugarcane bacilliform virus (ScBV), a badnavirus, consists of a circular dsDNA. The complete sequence of a cloned infective ScBV genome is reported here. The genome is 7568 bp in size and possesses a number of features suggesting that ScBV is a pararetrovirus. A tRNA(Met)-binding site that may serve as a primer for minus-strand synthesis is present. The plus-strand of the ScBV genome contains three open reading frames (ORFs) which are capable of encoding proteins with calculated M(r) values of 22K, 13K and 215K. The 215K protein has regions with similarity to the RNA-binding domains, aspartic proteases and replicases of retro-elements. In addition, the 215K protein also has a region with restricted similarity to the intercellular transport proteins of plant viruses. Comparisons with the other sequenced badnaviruses, Commelina yellow mottle (CoYMV) and rice tungro bacilliform (RTBV) viruses, indicate that the arrangement of the ORFs in these viruses is conserved. Located next to the putative RNA-binding domain is a cysteine-rich region that is unique to the badnaviruses. When the molecular relationships of a portion of the reverse transcriptases of plant pararetroviruses were determined, two badnaviruses, CoYMV and ScBV, form one distinct cluster, whereas three caulimoviruses, cauliflower mosaic virus, carnation etched ring virus and figwort mosaic virus, form a second cluster. The badnavirus RTBV and the caulimovirus soybean chlorotic mottle virus occupy intermediate positions between the clusters. When introduced by Agrobacterium-mediated inoculation, a construct containing 1.1 copies of the cloned ScBV genome is infectious to both rice and banana. PMID- 8423448 TI - Pseudorecombination between infectious cloned DNA components of tomato mottle and bean dwarf mosaic geminiviruses. AB - A newly described whitefly-transmitted geminivirus infecting tomato plants in Florida induces yellow mottling symptoms on leaves, and stunted and distorted growth. The DNA-A and DNA-B components were cloned from extracts of field infected tomato tissue; excised monomers or uncut tandem dimers of these clones were infectious when co-inoculated on to Nicotiana benthamiana by rub inoculation. Tomato plants inoculated directly with the DNA-A and DNA-B dimers, or indirectly by sap or graft transmission from N. benthamiana plants previously infected with the dimers, developed symptoms similar to those observed in field infected plants. This tomato geminivirus is different from previously characterized geminiviruses, and has been named tomato mottle geminivirus (ToMoV). DNA sequence comparisons revealed that ToMoV is closely related to bean dwarf mosaic geminivirus (BDMV) and abutilon mosaic geminivirus. Infectious pseudorecombinants were made by exchanging the cloned infectious DNA components of ToMoV and BDMV and inoculating N. benthamiana plants. The presence of the inoculated DNA components in systemically infected plants was confirmed by characterization of DNA-A and DNA-B fragments amplified by the polymerase chain reaction. This is the first report of pseudorecombination between two distinct geminiviruses. The implications of this finding in geminivirus evolution are discussed. PMID- 8423449 TI - Cell-to-cell transmission of human immunodeficiency virus infection induces two distinct phases of viral RNA expression under separate regulatory control. AB - A cell clone persistently infected with human T cell-lymphotrophic virus type IIIB (H3B cells) contained mainly the multiply spliced (2 kb) and singly spliced (4.3 kb) species of human immunodeficiency virus (HIV) RNA. When H3B cells were co-cultured with susceptible HUT78 cells, cell fusion occurred within 4 h of cell mixing and was accompanied by a marked increase of the unspliced full-length (9.2 kb) HIV RNA. This first phase of viral RNA induction (4 to 12 h post-infection) was followed by a second phase of viral RNA synthesis from 24 h p.i. in which there were significant increases in all three species of HIV RNA. Reverse transcriptase (RT) inhibitors such as azidothymidine (AZT) at concentrations that abolished de novo HIV DNA synthesis, abolished the first phase but not the second phase of viral RNA synthesis in our model system. A comparable one-step cell-free virus infection showed a pattern of viral RNA synthesis similar to that of the cell-to-cell transmission of infection. However, viral RNA synthesis following cell-free virus infection was totally inhibited by RT inhibitors. The early phase (4 to 12 h) expression of 9.2 kb HIV RNA is likely to use newly synthesized HIV DNA as template; during this phase, HIV RNA and DNA syntheses occur simultaneously, with each process being dependent on the other for maximal yield. During the later (24 to 48 h) phase, all three HIV RNA species may be transcribed at least in part from proviral DNA from the original donor cells. This later phase may provide one of the mechanisms for natural spread of virus to new cells and for enhanced viral gene expression in vivo, despite the presence of AZT. PMID- 8423450 TI - Viable double vaccinia virus recombinants with the non-inducible phage T7 expression system. AB - Double vaccinia virus recombinants expressing both the T7 RNA polymerase gene, controlled by a weak early poxvirus PF promoter, and the Escherichia coli beta galactosidase gene, controlled by the phage T7 promoter, have been obtained. The viability of the double recombinants depended on the T7 RNA polymerase expression level. If the T7 RNA polymerase gene was inserted into a recombinant already containing the beta-galactosidase gene, the efficiency of formation of the double recombinants was significantly higher compared to that for the reverse insertion order. The negative effect of the phage T7 terminator on beta-galactosidase expression in cells infected with the recombinant viruses has been shown. The dynamics and levels of beta-galactosidase formation by different vaccinia virus recombinants have been studied. PMID- 8423451 TI - Characterization of African horsesickness virus serotype 4-induced polypeptides in Vero cells and their reactivity in Western immunoblotting. AB - The structural and non-structural proteins induced by African horsesickness virus serotype 4 (AHSV-4) in infected Vero cells were analysed by SDS-PAGE. Twenty-two virus-induced polypeptides were detected in infected cells by comparison with the polypeptides of mock-infected cells, of which four major (VP2, VP3, VP5 and VP7) and three minor (VP1, VP4 and VP6) structural proteins and four non-structural proteins (P58, P48, P21 and P20) were shown to be virus-coded, as deduced from electrophoretic and antigenic studies of purified virions and infected cells. The proteins that elicit the major antibody responses both in vaccinated and naturally or experimentally infected horses were shown to be three structural proteins, VP2, VP5 and VP7, and the four major non-structural proteins, P58, P48, P21 and P20, as deduced by radioimmunoprecipitation and immunoblotting assays. The cross-reactivity between AHSV-4 and sera obtained from horses experimentally infected with seven other serotypes was also determined. The results showed that VP5, VP7, P48, P21 and P20 are conserved and can be used to diagnose the infection of any of these eight serotypes. PMID- 8423452 TI - Immunoreactivity and protective effects in mice of a recombinant dengue 2 Tonga virus NS1 protein produced in a baculovirus expression system. AB - The double-stranded replicative form of dengue 2 virus (DEN-2) RNA (Tonga strain) was used as a substrate to produce DNA clones of the NS1-NS2A genes via reverse transcriptase synthesis of full length cDNA followed by polymerase chain reaction amplification of the NS1-NS2A region. Products were cloned into pTZ18R for sequencing and into baculovirus for expression studies. The deduced amino acid sequence of the NS1-NS2A was almost identical to that of the S1 attenuated strain of DEN-2 Puerto Rico 159, differing in only four amino acids. The NS1 protein expressed in insect cells [Spodoptera frugiperda (Sf-9)] from the baculovirus recombinant was indistinguishable from authentic NS1 of DEN-infected Aedes albopictus cells in glycosylation, dimerization, cellular presentation and antigenicity. Mice injected with the expressed protein developed NS1-specific complement-fixing antibodies and were partially protected against neurological residua after intracranial challenge. The protective effect was mouse strain- and gender-specific. PMID- 8423453 TI - Expression of Bombyx mori cytoplasmic polyhedrosis virus polyhedrin in insect cells by using a baculovirus expression vector, and its assembly into polyhedra. AB - A cDNA encoding the cytoplasmic polyhedrin of Bombyx mori cytoplasmic polyhedrosis virus (BmCPV) strain H was introduced into an improved baculovirus expression vector which can be utilized to express foreign genes in the Spodoptera frugiperda cell line IPLB-SF-21AE (Sf21 cells) and the B. mori cell line BmN. A recombinant virus produced large, cubic inclusion body-like structures in infected Sf21 and BmN cells. Western blot analysis showed that these structures were BmCPV polyhedra. This result suggested that the supramolecular assembly of BmCPV polyhedrin is responsible for its properties. PMID- 8423455 TI - Genome analysis of adenovirus 4 isolated over a six year period. AB - Genome analysis was carried out on 74 adenovirus 4 (Ad4) isolates from patients in Manchester between 1984 and 1989. Most of the isolates were associated with conjunctivitis. Of the 74 isolates studied, 51 were Ad4a and 10 were Ad4p (the prototype strain). The remaining isolates consisted of two new genome types we have designated Ad4a2 (10 isolates) and Ad4a3 (3 isolates). Most of the genome types co-circulated during the period of study. The Bst E II and Xho I restriction maps of the new variants are presented and compared with those of Ad4p. We are unable to associate genome types with particular clinical presentations. PMID- 8423454 TI - Secular trend and geographical variation in hepatitis A infection and hepatitis B carrier rate among adolescents in Taiwan: an island-wide survey. AB - During the last two decades an economic boom has occurred in Taiwan, a region where the prevalence of both hepatitis A and B virus infection was formerly very high. To examine the impact of socioeconomic developments on the secular trend and geographical variation in hepatitis A and B virus infection, 875 adolescents selected randomly from 20 junior high schools were studied. Serum samples collected from the subjects were tested for hepatitis A antibody (anti-HAV) and hepatitis B surface antigen (HBsAg) by enzyme immunoassay using commercial reagents. The anti-HAV prevalence increased from northern through central to southern Taiwan; the prevalence was highest in aboriginal townships and lowest in metropolitan precincts. This striking variation in anti-HAV prevalence in different geographical locations and at different urbanization levels remained significant in multiple logistic regression analysis. The HBsAg prevalence was significantly higher in aboriginal townships than in rural and urban townships and metropolitan precincts. In addition, HBsAg prevalence was related inversely to the number of physicians per 1,000 population. The prevalence of both anti-HAV and HBsAg declined significantly during the last decade in Taipei City and County. The decrease in anti-HAV prevalence may be due to improvements in environmental hygiene, water supply, and food sanitation, while the decline in the HBsAg carrier rate may result from the use of disposable needles and syringes as well as screening for HBsAg in blood banks. PMID- 8423456 TI - Comparative study of human and rabbit cell infection with cell-free HTLV-I. AB - Infection of human and rabbit cells with cell-free HTLV-I was studied by PCR analysis. Both human and rabbit PBL were infected similarly by cell-free virus of both human and rabbit cell origin. Cells were infected with the cell-free virus without prior treatment and regardless of the concentration of the culture supernatant containing the virus. Human and rabbit cell lines were also infected similarly by the cell-free virus, the proviral DNA persisting for more than two months. The culture supernatants of HTLV-I-producing cells could thus be a potential cause of laboratory infections. PMID- 8423458 TI - Motor neuron disease and past poliomyelitis. Geographic study in Hokkaido, the northern-most island of Japan. ALS Study Group. AB - We carried out a survey on the cases of motor neuron disease (MND) occurring in Hokkaido between 1980 and 1989 in order to establish whether there is a relationship between MND and poliomyelitis. Hokkaido, the northern-most island of Japan, was one of the most affected areas during the poliomyelitis epidemic of the late 1940s to 1950s. We ascertained 389 cases of MND for these 10 years and 2,171 cases of paralytic poliomyelitis for the period 1949-1958, and analysed their geographic distributions. No significant correlation was found between MND and poliomyelitis in their geographic distributions. PMID- 8423457 TI - Chronic progressive sensory ataxic neuropathy: clinicopathological features of idiopathic and Sjogren's syndrome-associated cases. AB - Eleven patients with chronic progressive sensory ataxic neuropathy were examined clinicopathologically. Three cases were associated with primary Sjogren's syndrome (SS-SAN) and the others were considered to be idiopathic (ISAN). The major clinical symptom in both was loss of proprioceptive and kinesthetic sensation with some impairment of superficial sensation, with multifocal and asymmetrical distribution and progression. The truncal and trigeminal nerves were frequently involved. The motor system was substantially preserved. These somatic sensory and motor symptoms did not differ between ISAN and SS-SAN, but autonomic nervous system signs were more frequent in SS-SAN. Polyclonal elevations of serum IgG and/or IgA were seen in 8 patients. One autopsied case with ISAN combined with previous reports suggested that systemic T- and B-cell infiltration into the nervous tissues, as well as a wide variety of the visceral organs, may be a common finding in ISAN and SS-SAN, and could participate in the cause of this neuropathy and polyclonal hypergammaglobulinaemia. PMID- 8423459 TI - The use of monoclonal antibodies in diagnostic tests for Becker and Duchenne muscular dystrophy. AB - Monoclonal antibodies recognizing different epitopes of dystrophin have now been widely applied in diagnostic tests for Duchenne and Becker muscular dystrophy (BMD). The preservation of the C-terminus in BMD patients necessitates the routine use of one antibody raised against this region. Additional antibodies against deletion-prone regions of dystrophin further increase the diagnostic power of the tests. We describe two cases which illustrate that the combined use of different antibodies is necessary to avoid misdiagnoses of BMD at the protein level. PMID- 8423461 TI - Visuo-spatial neglect: a new copying test to assess perceptual parsing. AB - A new copying test has been devised to explore different forms of perceptuo-motor impairment in visuo-spatial neglect. The stimuli are constructed so that one hypercomplex figure can be transformed into two complex figures by deletion of part of the original drawing. Task performance by five patients with left neglect after right hemisphere damage is reported. Their copies illustrate the varied spatial "reference frames" within which visual neglect can be manifest. PMID- 8423460 TI - The dynamics of multiple sclerosis. The Charcot Lecture. AB - The history of our understanding of the pathogenesis and pathophysiology of multiple sclerosis are reviewed in the context of Charcot's contribution. The implications for treatment of the new knowledge gained from studies during life of pathology and pathogenesis (by MRI) and pathophysiology (by evoked potentials) are reviewed. PMID- 8423462 TI - Magnetic resonance imaging in patients with multiple sclerosis and spinal cord involvement: 28 cases. AB - In patients with clinically isolated spinal disease, magnetic resonance imaging (MRI) provides a non-invasive method of detecting surgically treatable causes and is also useful in detecting asymptomatic brain lesions where the cord syndrome is due to multiple sclerosis (MS). We report the findings of spinal and brain MRI in 28 patients with spinal cord disorder due to MS. It was possible to detect intrinsic plaques reliably in the majority of patients (60.7%) with clinical findings of spinal cord MS. The results of MRI are compared with the clinical status and with cerebrospinal fluid findings and evoked potentials. PMID- 8423463 TI - In vivo relationship of interleukin-2 and soluble IL-2 receptor to blood-brain barrier impairment in patients with active multiple sclerosis. AB - Interleukin (IL)-2 has well-recognized effects on cerebral endothelial cells and, therefore, may mediate disruption of the blood-brain barrier in patients with multiple sclerosis (MS). To evaluate the in vivo relationship of the IL-2 system to blood-brain barrier impairment in MS, levels of IL-2 and soluble IL-2 receptors (sIL-2R) in cerebrospinal fluid (CSF) and serum samples from 50 patients with active MS and 49 controls were correlated with values of the CSF to serum albumin ratio. Intrathecal levels of IL-2 and sIL-2R were significantly higher in MS compared with the control groups and correlated with albumin ratios in MS patients. Intrathecal levels of IL-2 and sIL-2R also correlated with the degree of barrier damage in these patients. It is suggested that intrathecal levels of IL-2 and sIL-2R are related to barrier impairment in MS and may be important in understanding some of the pathological changes of this condition. PMID- 8423464 TI - Magnetic resonance imaging of muscle and brain in myotonic dystrophy. AB - Myotonic dystrophy (MD) is characterized by myotonia, weakness and extramuscular symptoms, including intellectual impairment. We performed magnetic resonance imaging (MRI) of brain and muscle in 25 MD patients: 81% had cerebral atrophy (severe in 36%); 68% had focal white matter lesions, which were large and multiple in 27%. Brain MRI findings correlated with mental impairment; the severity of both correlated with disease duration. Changes in brain and muscle MRI were progressive with time, but independent of each other. Muscle MRI findings were fatty degeneration and loss of bulk. In the calves, the medial gastrocnemius muscles were involved earliest and the posterior tibial muscles relatively spared. In the thighs the vastus muscles were damaged most often and the rectus femoris least. Focal muscle damage was efficiently visualized, sometimes preceding clinical detection. Muscle MRI was less sensitive than conventional methods for early diagnosis, but ideal for follow-up, owing to its non-invasiveness and examiner-independence. PMID- 8423465 TI - Activation of single neurons in the rat nucleus accumbens during self-stimulation of the ventral tegmental area. AB - Single neurons (n = 76) were recorded in the nucleus accumbens septi (NAS) of rats self-stimulating the ipsilateral medial forebrain bundle (MFB) at the level of the ventral tegmental area (VTA). Responses evoked by rewarding trains of stimulus pulses fell into five categories. The first category (40% of the sample) was characterized by a single discharge at invariant latency in response to individual pulses of the train, and hence was termed "tightly time locked" (TTL). Two TTL neurons were collision tested, and both showed collision, suggesting that self-stimulation of the VTA may involve antidromic, and thus direct, activation of a substantial number of NAS axons. The second category (26%) was characterized by discharges that varied in latency from pulse to pulse and hence was termed "loosely time locked" (LTL). Responses of the remainder of the sample showed no coupling to individual pulses but were categorized based on general firing patterns during the train: excited (7%), inhibited (4%), and no change (23%). Irrespective of category, immediately after the self-stimulation session, the likelihood of evoked discharge at monosynaptic latency by single pulse stimulation of the ipsilateral fimbria was reduced (relative to pre-session level), concurrent with elevations in mean firing rate and motor activity. NAS neurons thus exhibit vigorous activation, apparently both antidromically and orthodromically, in response to VTA self-stimulation. The responses of certain LTL and TTL neurons increased as a function of pulse number in the train, suggestive of integrative mechanisms important for brain stimulation reward. Conduction velocities of directly activated (TTL) axons (0.41-0.65 m/sec) were slower than those previously reported for first-stage, reward-relevant axons. Nonetheless, an implication of direct activation of NAS (and other MFB) axons is that rewarding stimulation triggers action potentials that could invade all axonal branches, including those between the stimulation site and the soma, and send synaptic signals to target neurons. Such signals from NAS neurons could contribute to the increased motor behavior accompanying MFB self-stimulation, and/or could interact with dopamine-mediated signals projected to the NAS from reward circuitry. PMID- 8423466 TI - Novel opioid binding sites associated with nuclei of NG108-15 neurohybrid cells. AB - Nuclear opioid binding sites have been discovered in NG108-15 neurohybrid cells. Marker enzyme analyses as well as electron and fluorescence microscopy studies attested to the high degree of purity of the nuclear preparations. Immunohistochemical studies on cryostat sections of NG108-15 cells with an antibody to the opioid receptor corroborated a nuclear localization. 3H-[D-Pen2,D Pen5]enkephalin (3H-DPDPE), 3H-[D-Ala2,D-Leu5]enkephalin (3H-DADLE), and 3H diprenorphine binding parameters, Kd and Bmax values, and heterologous competition binding and stereospecificity data satisfied criteria for the presence of delta-opioid sites in purified nuclear preparations. Neither mu-([D Ala2,mephe4,gly-ol5] enkephalin), dihydromorphine, nor kappa-(U69593) specific binding was detectable in purified nuclear preparations. Rates of association and dissociation of 3H-[D-Ser2,L-Leu5]enkephalyl-Thr were comparable to values obtained previously for opioid receptors. Opioid binding was also shown in subnuclear preparations from NG108-15 cell cultures. Agonists, 3H-DADLE and 3H DPDPE, bind with high affinity to nuclear membranes and with lower affinity to chromatin. In contrast, partial agonist 3H-diprenorphine high-affinity binding sites were predominant in chromatin, while low-affinity binding was found in the nuclear membrane. Accordingly, 5'-guanylylimidodiphosphate sensitivity of 3H DADLE binding was detected in nuclear membranes but not in chromatin. Both agonist and partial agonist opioid binding to nuclear membrane and chromatin were abolished upon cycloheximide treatment of NG108-15 cells. Taken together, the results suggest that NG108-15 cells contain newly synthesized GTP binding regulatory protein (G-protein)-coupled delta-opioid receptors in nuclear membranes and uncoupled opioid binding sites in chromatin. PMID- 8423467 TI - Cell-cell interactions during the migration of an identified commissural growth cone in the embryonic grasshopper. AB - One of the fascicles of the posterior commissure of the embryonic grasshopper is pioneered by an individually identifiable neuron named Q1. Q1 initially grows along a longitudinal pathway established by another pioneer neuron, MP1, and then crosses to the midline, where it meets and fasciculates with the axon of the contralateral Q1. The Q1 growth cone follows the contralateral Q1 axon to the contralateral longitudinal pathway, where it then fasciculates with axons of the MP1/dMP2 fascicle. In this work, we have identified a small set of early neurons that Q1 could use as guidance cues while negotiating its way along a specific and stereotyped pathway to the midline. Furthermore, we have observed characteristic morphological changes in the Q1 growth cone that could indicate responses to changing adhesivity in the substrates it contacts. We have also quantified the pattern of dye coupling between neurons in this system. Most of the neurons to which Q1 becomes coupled retain a strong, consistent pattern of dye coupling that shows no recognizable variation at times when growth cones are making pathway decisions. However, we have found one clear instance of transient, site-specific dye coupling between the Q1 growth cone and the ipsilateral MP1 soma. The timing and pattern of dye coupling in this system suggest that dye coupling may play a role in synchronizing the initiation of axon outgrowth among a small population of neurons. Although dye coupling may not play a direct role in neuronal pathfinding, it may exert a profound indirect influence on neuronal interactions by regulating the timing of axon outgrowth. PMID- 8423468 TI - Growth cone dynamics during the migration of an identified commissural growth cone. AB - We have used time-lapse video microscopy to study the behavior of a neuron, Q1, that pioneers the posterior commissure of the embryonic grasshopper. Our goal is to use time-lapse video as a tool to acquire a precise picture of normal development over time, and thereby identify stereotypic activities that might indicate important interactions necessary for proper formation of the commissure. We have identified specific and reproducible behaviors that suggest the presence of underlying cellular interactions that may play a role in pathfinding. In particular, the Q1 growth cone undergoes several morphological changes as it contacts the midline. As a commissural neuron, the midline may be a target in its outgrowth; Q1's typical response upon contacting the midline with its filopodia, however, is a rapid retraction. This inhibitory reaction can be overridden by contact with filopodia of its contralateral homolog. Q1's growth cone can translocate across the midline at an accelerated rate by a process resembling "filopodial dilation" (O'Connor et al., 1990) once the two Q1 growth cones meet. Ablation of the contralateral Q1 blocks Q1's advance across the midline. We have also analyzed in detail the behavior of individual filopodia to identify behavioral differences that could indicate differences in substrate adhesivity. Except for instances of filopodial dilation seen only at the midline, we found no significant asymmetries in rates of filopodial extension and retraction, or in the survival times of individual filopodia. We suggest that either the adhesive signal used by Q1 is relatively weak, requiring the integration of many adhesive interactions by many filopodia to be resolved, or the guidance cues may not be adhesive in nature. PMID- 8423469 TI - Rapid evolution of the visual system: a cellular assay of the retina and dorsal lateral geniculate nucleus of the Spanish wildcat and the domestic cat. AB - The large Spanish wildcat, Felis silvestris tartessia, has retained features of the Pleistocene ancestor of the modern domestic cat, F. catus. To gauge the direction and magnitude of short-term evolutionary change in this lineage, we have compared the retina, the optic nerve, and the dorsal lateral geniculate nucleus (LGN) of Spanish wildcats and their domestic relatives. Retinas of the two species have the same area. However, densities of cone photoreceptors are higher in wildcat--over 100% higher in the area centralis--whereas rod densities are as high, or higher, in the domestic lineage. Densities of retinal ganglion cells are typically 20-100% higher across the wildcat retina, and the total ganglion cell population is nearly 70% higher than in the domestic cat. These differences are confined to the populations of beta and gamma retinal ganglion cells. In contrast, the population of alpha cells is almost precisely the same in both species. The wildcat LGN is much larger than that of the domestic cat and contains approximately 50% more neurons. However, cell size does not differ appreciably in either the retina or LGN of these species. The differences in total numbers of ganglion cells and LGN neurons correspond neatly to the overall decline in brain size in the domestic lineage and to allometric predictions based on average species differences in body size. We suggest that an increase in the severity of naturally occurring cell death is the most plausible mechanism that can account for the rapid evolutionary reduction in cell populations in this feline lineage. PMID- 8423470 TI - Simultaneous pontine and basal forebrain microinjections of carbachol suppress REM sleep. AB - This study was performed to test the hypothesis that cholinoceptive basal forebrain systems can significantly influence cholinoceptive pontine mechanisms known to be important for generating rapid eye movement (REM) sleep. This hypothesis was examined by microinjecting the cholinergic agonist carbachol or saline (vehicle control) into the pons, the basal forebrain, or simultaneously into the pons and basal forebrain, while quantifying the effects on sleep and wakefulness in unanesthetized, chronically instrumented cats. All microinjections were made during wakefulness and were followed by 2 or 4 hr of recording. Polygraphic records were scored for wakefulness, non-REM sleep, REM sleep, and the REM sleep-like state evoked by pontine administration of carbachol (DCarb). Dependent variables quantified following each microinjection included the percentage of recording time spent in each state, the latency to onset of non REM, REM, and DCarb, the number of episodes per hour of each state, and the duration of the longest episode of each state. A total of 149 microinjections were made into 15 forebrain and 11 pontine sites in eight cats. Basal forebrain administration of carbachol significantly increased wakefulness. Pontine microinjection of carbachol produced a state that polygraphically and behaviorally resembled REM sleep. This REM sleep-like state occurred in amounts significantly greater than natural REM sleep. Pontine carbachol also significantly decreased wakefulness and non-REM sleep. Simultaneous injection of carbachol into the pons and basal forebrain enhanced REM sleep, but the magnitude of this enhancement was significantly less than the increase in REM sleep evoked by carbachol injection into the pons alone. The results show that cholinoceptive regions of the basal forebrain can increase wakefulness and reduce the ability of pontine carbachol to evoke the REM sleep-like state. These findings suggest that basal forebrain administration of carbachol activates an arousal-generating system that can successfully compete with the powerful cholinergic REM sleep generating system of the pons. PMID- 8423471 TI - Afferent spontaneous electrical activity promotes the survival of target cells in the developing retinotectal system of the rat. AB - This study was undertaken to investigate the role of afferent spontaneous electrical activity in regulating death of target cells in the developing mammalian visual system. We show here that naturally occurring cell death in the rat superior colliculus is greatly augmented when the spontaneous firing of retinal ganglion cells is transiently blocked with TTX. An increased number of dying cells is already observed after 1 hr of afferent blockade. A 50% increase of cell death is reached after 3 hr of blockade, an effect that closely parallels increased cell death caused by eye enucleation after similar intervals of time. These results suggest that, during development, input cells exert a trophic action on target cells, which is prevented by silencing input electrical activity. A likely explanation of this effect is that the spontaneous firing of input cells causes the release by afferent fibers of a trophic agent promoting the survival of target cells. PMID- 8423472 TI - Damage to the perirhinal cortex exacerbates memory impairment following lesions to the hippocampal formation. AB - Recent work has been directed at identifying the critical components of the medial temporal lobe that, when damaged, produce severe memory impairment. The H+A+ lesion includes the hippocampal formation, the amygdala, and the adjacent entorhinal, parahippocampal, and perirhinal cortices. A more restricted medial temporal lobe lesion that includes the hippocampal formation and parahippocampal cortex (the H+ lesion) produces less severe memory impairment. Previous work demonstrated that extending the H+ lesion forward to include the amygdala did not exacerbate the impairment. Here, we tested the hypothesis that extending the H+ lesion forward to include the perirhinal cortex (the H++ lesion), but sparing the amygdala, should produce a more severe memory impairment and one that would approximate the level of memory impairment associated with the H+A+ lesion. Monkeys with the H++ lesion were severely impaired on two of three amnesia sensitive tasks (delayed nonmatching to sample and delayed retention of object discrimination). On the third amnesia-sensitive task (concurrent discrimination learning), two of the monkeys in the H++ group obtained poorer scores than all seven normal monkeys, although the overall group comparison was not significant. The memory impairment following H++ damage was more severe overall than the impairment associated with the H+ lesion and approached the level of impairment associated with the H+A+ lesions. Quantitative measurement of damage in each anatomical component of the lesion indicated that the perirhinal cortex was the only brain region that was more extensively damaged in the H++ group than in the H+ group. These findings emphasize the importance of the perirhinal cortex in the anatomy of the medial temporal lobe memory system. PMID- 8423473 TI - Time course of extracellular dopamine and behavioral sensitization to cocaine. I. Dopamine axon terminals. AB - Repeated administration of cocaine to rodents produces a progressive augmentation in motor activity known as behavioral sensitization. By using microdialysis in the ventral striatum, some studies have found that the development of behavioral sensitization is associated with a similar augmentation in dopamine release, while others have not. It was postulated that differences in doses and withdrawal periods may account for the discrepancies between studies. Rats were behaviorally sensitized to daily peripheral injections using two cocaine treatment regimens (15 mg/kg, i.p. x 5 d or 30 mg/kg, i.p. x 5 d). Using in vivo microdialysis in the ventral striatum, the effect of acute cocaine (15 mg/kg, i.p.) on extracellular dopamine content and motor behavior was examined at various times after discontinuing daily treatments. Twenty-four hours after discontinuing the low dose of daily cocaine, the increase in motor activity and extracellular dopamine elicited by an acute cocaine challenge was significantly elevated. In contrast, following the higher daily treatment regimen there was a significant augmentation in motor activity, but the increase in extracellular dopamine produced by cocaine was significantly reduced. When rats were challenged 10-14 d after discontinuing either dosage regimen of daily cocaine, the increase in both motor activity and extracellular dopamine was augmented. In general, the increase in extracellular dopamine by an acute cocaine challenge increased over time when rats were challenged between 1 and 22 d after discontinuing daily cocaine. Basal concentrations of extracellular dopamine were determined by measuring the in vivo flux of dopamine across the dialysis membrane, and there was no significant difference at 24 hr or 2 weeks following the last daily injection of saline or cocaine. It is concluded that behavioral sensitization to cocaine is generally associated with an augmentation in extracellular dopamine in the ventral striatum, but that high doses of daily cocaine produce apparent tolerance to the augmentation in extracellular dopamine during the early withdrawal period. PMID- 8423474 TI - Initial tract formation in the mouse brain. AB - Mouse embryos from embryonic days 8.5-10.5 (E8.5-E10.5) were fixed and labeled with an antibody to neuron-specific class III beta-tubulin (Moody et al., 1987; Lee et al., 1990a,b) to reveal the first neurons, axons, and tracts in the brain. They were studied in whole-mounts and in light microscopic sections. Some conclusions were checked by labeling tracts in older embryos (E11.5 and E12.5) with the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine. The first immunoreactive cells appeared at E8.5, prior to neural tube closure, in the neural plate immediately caudal to the optic vesicle. Cells along the dorsal midline of the mesencephalon issued the first axons, on E9.0; the cells were the mesencephalic nucleus of the trigeminal nerve, and the axons formed its descending tract. The tract reached the level of the trigeminal ganglion by E10.0 but did not enter the ganglion until after E12.5. On E9.5, the number of labeled cells and axons in the alar plate of the presumptive diencephalon and mesencephalon had increased substantially, and many of the rostral ones coursed into the basal plate to enter longitudinal tracts there. Two tracts originated from cells in the basal plate: the tract of the postoptic commissure (from the base of the optic stalk to the level of the cephalic flexure) and the medial longitudinal fasciculus (from the level of the cephalic flexure caudally through the mid and hind-brains). By E10.0, a small mammillotegmental tract paralleled the tract of the postoptic commissure, but immunolabeling was so widespread that discrete tracts were impossible to discern in the presumptive diencephalon and mesencephalon. The more rostral regions remained lightly labeled. In the cerebral vesicle, the presumptive cerebral cortex, the first immunoreactive cells appeared at E10.0; they had multiple processes oriented parallel to the pia, and were identified as the Cajal-Retzius cells. By E10.5, no tracts had formed in the cerebral vesicle. All the tracts formed by E10.0 were superficial, in the subpial lamina. Those that can be identified in the adult brain are very deep structures. These results are compared with previous descriptions of the embryonic brains of amphibians, fish, birds, and other mammals, including humans. PMID- 8423475 TI - Brain glia release factors with opposing actions upon neuronal survival. AB - Microglia and astroglia have been thought to govern the survival of neurons after damage to the CNS. To investigate these putative glia-neuron relationships, we examined microglia and astroglia secretion products for effects upon growth of cultured neurons. Activated microglia secrete small neurotoxic factors (< 500 Da), while astroglia constitutively release proteins (> 10 kDa) that promote neuronal growth. Proteins released from astroglia, moreover, attenuate microglial toxicity, suggesting that different glial populations have opposing actions upon neuronal survival. Further study shows that neurotoxins from microglia are heat stable, protease-resistant molecules with biologic activities blocked by NMDA receptor antagonists. Microglial factors, although toxic for chick ciliary neurons and rat spinal cord neurons, did not reduce numbers of oligodendroglia, astroglia, or Schwann cells in culture. The microglial neurotoxins can be distinguished from cytokines, from free radical intermediates, from the excitatory amino acids glutamate or aspartate, and from the NMDA receptor mediated toxin quinolinic acid. We propose that secretion products from reactive microglia, but not astroglia, endanger surviving neurons after CNS injury by release of a novel class of neuron-killing molecules. PMID- 8423476 TI - Directed expression of an oncogene to the olfactory neuronal lineage in transgenic mice. AB - The mammalian olfactory system provides a useful model to understand the cellular and molecular mechanisms governing the development of the nervous system. The olfactory neuroepithelium undergoes continual turnover in the adult animal, resulting in a neural tissue containing cells at various stages of neurogenesis. We have generated a transgenic mouse line to examine the effects of directed expression of an oncogene within the olfactory neuronal lineage. A hybrid oncogene was constructed utilizing the regulatory elements for the olfactory marker protein gene to direct the olfactory neuronal-specific expression of simian virus 40 T-antigen, a potent oncogene. The resulting transgenic mouse line expressed T-antigen only in olfactory neurons. Ten-month-old transgenic mice displayed significant hypoplasia of the neuronal elements in the olfactory neuroepithelium. The transgenic mice developed neuroblastomas of olfactory neuronal origin at a low frequency. Distinct clonal lines were derived from the primary culture of the tumor. GAP-43, a growth-associated neuronal marker, was expressed by some of the cell lines. One of the cell lines, 2.2, appeared to be responsive to neurotrophic effects from the presumptive target tissue, the olfactory bulb. PMID- 8423477 TI - Prenatal development of excitability in cat retinal ganglion cells: action potentials and sodium currents. AB - The development of precise retinofugal projections is dependent on activity mediated events, but as yet nothing is known about the ontogeny of excitable membrane properties in retinal ganglion cells (RGCs). In order to begin to understand how functional maturity is attained in these neurons, whole-cell patch clamp recordings were obtained from acutely dissociated RGCs of fetal and postnatal timed-pregnant cats. Current-clamp recordings revealed a pronounced developmental increase in the proportion of RGCs capable of generating action potentials. At embryonic day 30 (E30), 5 weeks before birth and during a time when RGCs are still being generated, electrical stimulation elicited spikes in only a third of the cells. None of these neurons were capable of multiple discharges in response to maintained depolarization. The proportion of spiking neurons increased during ontogeny, such that by E55 all RGCs could be induced to generate action potentials, with the majority manifesting repetitive spiking patterns. Application of tetrodotoxin abolished spike activity of all fetal RGCs, indicating that sodium-mediated action potentials are present very early in development. At the same time, voltage-clamp recordings revealed significant ontogenetic modifications in several key properties of the sodium currents (INa). These were (1) a twofold increase in Na current densities; (2) a shift in the voltage dependence of both activation and steady state inactivation: with maturity, sodium currents activate at more negative potentials, while steady state inactivation of INa occurs at less negative potentials; and (3) a decrease in decay time constants of the Na current, at membrane potentials negative to -15 mV. These developmental changes were largely restricted to the period of axon ingrowth (E30-E38), suggesting that maturation of INa is not the limiting factor for the onset of activity-dependent restructuring of retinofugal projections. PMID- 8423478 TI - The roles of sex, innervation, and androgen in laryngeal muscle of Xenopus laevis. AB - The relative contributions of innervation and androgen to three muscle fiber properties--twitch type, size, and number--were examined in the sexually dimorphic, androgen-sensitive laryngeal muscle of Xenopus laevis. In adults, the muscle contains all fast-twitch fibers in males and fast- and slow-twitch fibers in females; laryngeal muscle fibers are larger and more numerous in males than in females. Juvenile larynges are female-like in both sexes; male laryngeal muscle is subsequently masculinized by androgen secretion during postmetamorphic development. Because both laryngeal motor neurons and muscle fibers are androgen sensitive during masculinization, we examined the role of the nerve in androgen regulated muscle fiber development. Laryngeal muscle of male and female juvenile frogs was unilaterally denervated, and effects on muscle fiber type, size, and number were examined 4 weeks later. Half of the frogs received a dihydrotestosterone pellet at the time of denervation. Androgen treatment converts laryngeal muscle from mixed slow and fast to all fast twitch in both innervated and denervated muscle. Thus, the nerve is not required for androgen regulated fiber type expression in either sex. Denervation produces muscle fiber atrophy and androgen treatment induces muscle fiber hypertrophy in male and female larynx. Nerve and hormone effects are independent and additive; fiber size in androgen-treated denervated muscle is greater than in untreated innervated muscle, and fiber size in androgen-treated denervated muscle is smaller than in androgen-treated innervated muscle. There is no sex difference in the effects of innervation or androgen on fiber size. Denervation causes laryngeal muscle fiber loss in males but not in females. Androgen treatment protects male laryngeal muscle from denervation-induced fiber loss and causes fiber addition in innervated female laryngeal muscle. We conclude that there is a sexually dimorphic interaction between innervation and androgen in control of laryngeal muscle fiber number. PMID- 8423479 TI - The highly irregular firing of cortical cells is inconsistent with temporal integration of random EPSPs. AB - How random is the discharge pattern of cortical neurons? We examined recordings from primary visual cortex (V1; Knierim and Van Essen, 1992) and extrastriate cortex (MT; Newsome et al., 1989a) of awake, behaving macaque monkey and compared them to analytical predictions. For nonbursting cells firing at sustained rates up to 300 Hz, we evaluated two indices of firing variability: the ratio of the variance to the mean for the number of action potentials evoked by a constant stimulus, and the rate-normalized coefficient of variation (Cv) of the interspike interval distribution. Firing in virtually all V1 and MT neurons was nearly consistent with a completely random process (e.g., Cv approximately 1). We tried to model this high variability by small, independent, and random EPSPs converging onto a leaky integrate-and-fire neuron (Knight, 1972). Both this and related models predicted very low firing variability (Cv << 1) for realistic EPSP depolarizations and membrane time constants. We also simulated a biophysically very detailed compartmental model of an anatomically reconstructed and physiologically characterized layer V cat pyramidal cell (Douglas et al., 1991) with passive dendrites and active soma. If independent, excitatory synaptic input fired the model cell at the high rates observed in monkey, the Cv and the variability in the number of spikes were both very low, in agreement with the integrate-and-fire models but in strong disagreement with the majority of our monkey data. The simulated cell only produced highly variable firing when Hodgkin Huxley-like currents (INa and very strong IDR) were placed on distal dendrites. Now the simulated neuron acted more as a millisecond-resolution detector of dendritic spike coincidences than as a temporal integrator. We argue that neurons that act as temporal integrators over many synaptic inputs must fire very regularly. Only in the presence of either fast and strong dendritic nonlinearities or strong synchronization among individual synaptic events will the degree of predicted variability approach that of real cortical neurons. PMID- 8423480 TI - Orienting head movements resulting from electrical microstimulation of the brainstem tegmentum in the barn owl. AB - The size and direction of orienting movements are represented systematically as a motor map in the optic tectum of the barn owl (du Lac and Knudsen, 1990). The optic tectum projects to several distinct regions in the medial brainstem tegmentum, which in turn project to the spinal cord (Masino and Knudsen, 1992). This study explores the hypothesis that a fundamental transformation in the neural representation of orienting movements takes place in the brainstem tegmentum. Head movements evoked by electrical microstimulation in the brainstem tegmentum of the alert barn owl were cataloged and the sites of stimulation were reconstructed histologically. Movements elicited from the brainstem tegmentum were categorized into one of six different classes: saccadic head rotations, head translations, facial movements, vocalizations, limb movements, and twitches. Saccadic head rotations could be further subdivided into two general categories: fixed-direction saccades and goal-directed saccades. Fixed-direction saccades, those whose direction was independent of initial head position, were elicited from the midbrain tegmentum. Goal-directed saccades, those whose direction changed with initial head position, were elicited from the central rhombencephalic reticular formation and from the efferent pathway of the cerebellum. Particular attention was paid to sites from which fixed-direction saccadic movements were elicited, as these movements appeared to represent components of orienting movements. Microstimulation in the medial midbrain tegmentum elicited fixed-direction saccades in one of six directions: rightward, leftward, upward, downward, clockwise roll, and counterclockwise roll. Stimulation in and around the interstitial nucleus of Cajal (InC; a complete list of anatomical abbreviations is given in the Appendix) produced ipsiversive horizontal saccades. Stimulation in the ventral InC and near the dorsal and medial edges of the red nucleus produced upward saccades. Stimulation in the reticular formation near the lateral edge of the red nucleus produced downward saccades. Stimulation in the ventromedial central gray produced ipsiversive roll saccades. The metrics and kinetics of fixed-direction saccades, but not their directions, could be influenced by stimulation parameters. As such, direction was an invariant property of the circuits being activated, whereas movement latency, duration, velocity, and size each demonstrated dependencies on stimulus amplitude, frequency, and duration. The data demonstrate directly that at the level of the midbrain tegmentum there exists a three-dimensional Cartesian representation of head-orienting movements such that horizontal, vertical, and roll components of movement are encoded by anatomically distinct neural circuits.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423481 TI - Sound-localization deficits induced by lesions in the barn owl's auditory space map. AB - Barn owls possess a two-dimensional map of auditory space. The map appears in its final form in the external nucleus of the inferior colliculus (ICx) and is projected from there to the optic tectum (OT). To determine the role of the map in ICx or its projected version in OT in the localization of acoustic stimuli, head movements of three adult owls were recorded before and after lesioning parts of the map either in ICx or in OT. Small electrolytic lesions caused sound localization deficits that were characterized by failures to turn, turns away from the sound source, increased response latencies, and reduced turning amplitudes. These deficits occurred at azimuths expected from the physiological data obtained at the lesion site before passing current. They extended over an azimuthal range of some 20 degrees. Thus, this is the first unambiguous demonstration, for sound localization, of a deficit covering a well circumscribed area within an auditory hemifield. Since the major lesion deficits were restricted to one hemifield, a second lesion was made in the space map of the other side of the brain in each animal. The second lesion resulted in behavioral deficits qualitatively similar to those produced by the first lesion. In total, the study is based on nine lesions, three in each owl. Two lesions were sham lesions. These and two further lesions did not produce a measurable deficit. In four of the five remaining cases, in which a behavioral deficit was observed initially, the behavioral performance recovered with time. In two cases, the deficit disappeared completely. Although improvement occurred also in the remaining two cases, prelesion response amplitudes were not reached in one case, and response latency did not reach prelesion values in the other case. Because the behavioral deficits induced by the second lesions disappeared in the same way as did the deficits induced by the first lesions, the possibility is excluded that the animals learned to respond tactically, that is, that they learned to associate "unlocatable sounds" with the lesioned side. The deficits were also not due to a generalized motor impairment, because immediately after the lesion the animals responded as they did before the lesion to stimulation from outside the well circumscribed affected region of space. The possibility is discussed that plasticity after small central (neural) injuries is maintained longer in the lifetime of an animal than plasticity after peripheral (mechanical) manipulations. PMID- 8423482 TI - The expression of the protein p68/70 within the goldfish visual system suggests a role in both regeneration and neurogenesis. AB - Previous attempts to characterize the molecular events that support successful regeneration of axotomized goldfish retinal ganglion cells (RGCs) have led to the discovery of an acidic protein doublet in retina that displays an increased incorporation of 35S-methionine following axotomy, and is known to be axonally transported within the optic nerve. This protein is termed p68/70, reflecting its relative migration in 10% SDS-PAGE. In the present study, an affinity-purified polyclonal antibody to p68/70 (anti-p68/70) was developed and used to assess the species, tissue, and cellular distribution of p68/70. The antibody cross-reacted with homogenates of brain and other tissues from goldfish and closely related fish species. While each goldfish tissue tested expressed p68/70, the levels varied over a 30-fold range, with the highest amounts in brain, egg, and ovary. Immunolabeling of goldfish retina revealed prominent staining of RGC somata, dendrites, and axons. During regeneration, the immunoreactivity of the RGC somata and axons increased dramatically. Intense immunolabeling was also observed in the germinal neuroepithelial cells and rod precursors and in all retinal layers near the peripheral margin, in the region of recently differentiated neurons. In the tectum, the germinal zone was also highly labeled. The elevated expression of p68/70 in each of these areas known to mediate neuronal growth within the goldfish visual system suggests that p68/70 plays a role in axonal growth, regrowth, and possibly in neural development as well. PMID- 8423483 TI - Postural force fields of the human arm and their role in generating multijoint movements. AB - When a perturbation displaces the human hand from equilibrium, arm muscles respond by producing restoring forces. When a set of displacements are given at various directions from the same equilibrium position, the resulting restoring forces form a "postural force field." It is not known whether these postural forces are related to those generated when a reaching movement is executed. However, if a movement is a consequence of a shift of the equilibrium position of the hand toward the target, then, from the postural force field, predictions can be made regarding the nature of the elastic forces acting on the hand during the movement. We have taken the first steps in testing this hypothesis by measuring the postural force field of a subject's arm over relatively large distances, and comparing these forces with the static forces generated at the hand while the subject attempted a reaching movement. Using a robot manipulandum, the hand was displaced at various directions from an equilibrium position. The measured restoring forces were fitted to a nonlinear model to define a postural force field for that equilibrium position. This field was used to predict elastic forces generated when the subject attempted to move the manipulandum from a point on the circumference of a circle to a target at its center--the center corresponded to the equilibrium position at which the postural field was measured. In some of the movement trials, the manipulandum was locked during approximately the first 120 msec of the program for motion and the resulting static "evoked" forces measured. We found that (1) the evoked forces did not point to the target, but were a function of the configuration of the arm and rotated with the shoulder joint, and (2) the magnitude of the evoked forces varied systematically, even though the movements were of the same magnitude. These patterns were remarkably similar to those observed in the postural forces. Our results provide experimental evidence linking maintenance of posture in a multijoint system to that of generating a movement. The evidence is consistent with the hypothesis that the CNS programs a reaching movement by shifting the equilibrium position of the hand toward the target. PMID- 8423484 TI - Comparative characterization and autoradiographic distribution of neuropeptide Y receptor subtypes in the rat brain. AB - Some evidence has suggested the existence and differential distribution of neuropeptide Y (NPY) receptor subtypes in the mammalian brain (Dumont et al., 1990; Aicher et al., 1991). We now report on the extensive characterization and visualization of at least two classes of NPY receptor sites using a highly selective Y1 analog, [Leu31,Pro34]-NPY or [Pro34]-NPY, and a relatively specific Y2 competitor, NPY13-36. Autoradiographic studies using 125I-peptide YY (125I PYY) clearly reveal that the Y1 receptor subtype is most abundant in various cortical areas, the dentate gyrus of the hippocampal formation, the claustrum, and the reuniens nucleus of the thalamus. In most other regions, 125I-PYY binding is potently inhibited by increasing concentrations of either NPY2-36 or NPY13-36, suggesting a Y2-like profile. Furthermore, binding assays using homogenates from discrete brain regions clearly demonstrate that various NPY fragments and analogs compete for 125I-PYY labeling with profiles indicative of heterogeneity of NPY receptor subtypes, even in the presence of a selective Y1 blocker. Thus, it is likely that, in addition to the Y1 receptor, which is particularly concentrated in cortical areas, the rat brain is enriched with a receptor class (Y2) that can exist under high- or low-affinity states or with additional receptor subtypes that are recognized by 125I-PYY. These findings cannot be explained by the existence of the very recently reported Y3 receptor subtype, since PYY does not possess significant affinity to this site (Grundemar et al., 1991). Further experiments are currently in progress to determine the nature and functional significance of each of these NPY/PYY receptor sites. PMID- 8423485 TI - Plasticity in the frequency representation of primary auditory cortex following discrimination training in adult owl monkeys. AB - Previous studies have shown that the tonotopic organization of primary auditory cortex is altered subsequent to restricted cochlear lesions (Robertson and Irvine, 1989) and that the topographic reorganization of the primary somatosensory cortex is correlated with changes in the perceptual acuity of the animal (Recanzone et al., 1992a-d). Here we report an increase in the cortical area of representation of a restricted frequency range in primary auditory cortex of adult owl monkeys that is correlated with the animal's performance at a frequency discrimination task. Monkeys trained for several weeks to discriminate small differences in the frequency of sequentially presented tonal stimuli revealed a progressive improvement in performance with training. At the end of the training period, the tonotopic organization of Al was defined by recording multiple-unit responses at 70-258 cortical locations. These responses were compared to those derived from three normal monkeys and from two monkeys that received the same auditory stimuli but that were engaged in a tactile discrimination task. The cortical representation, the sharpness of tuning, and the latency of the response were greater for the behaviorally relevant frequencies of trained monkeys when compared to the same frequencies of control monkeys. The cortical area of representation was the only studied parameter that was correlated with behavioral performance. These results demonstrate that attended natural stimulation can modify the tonotopic organization of Al in the adult primate, and that this alteration is correlated with changes in perceptual acuity. PMID- 8423486 TI - Promoting breast-feeding among vulnerable mothers. PMID- 8423487 TI - Fundal height measurement. Part 2--Intra- and interexaminer reliability of three measurement techniques. AB - The intra- and interexaminer reliability of three fundal height measurement techniques (two tape measure and one caliper) was studied in a convenience sample of 60 pregnant women. The three measurements were obtained twice by two examiners. Four examiners with varying levels of clinical experience participated in the study; one examiner obtained measurements in all 60 cases and the other three examiners each obtained measurements in approximately one-third of the cases. Examiners were blinded to their own and to other examiners' measurements. The mean absolute differences between individual examiner's first and second measurements varied from 0.68 to 1.74 cm, the percentage of differences < or = 1.0 cm varied from 45% to 77.8%, and the maximal differences varied from 1.6 to 7.5 cm. The caliper technique consistently demonstrated the smallest mean absolute intraexaminer differences (0.68 to 1.39 cm) and the highest percentage of intraexaminer differences < or = 1.0 cm (55% to 77.8%). The differences between pairs of examiners' measurements were consistently greater than the differences between individual examiner's measurements: Mean absolute differences varied from 1.36 to 3.60 cm, the percentage of differences < or = 1.0 cm varied from 9.1% to 55.6%, and the maximal differences varied from 3.1 to 11.5 cm. Interexaminer differences were smallest for the tape "over-the-curve" measurements and the caliper measurements. These findings indicate that, to ensure maximal reliability, fundal height measurements should be obtained by the same clinician throughout pregnancy. Additionally, these findings indicate that calipers may be the most reliable method of obtaining fundal height measurements and that the accuracy of caliper measurements in identifying fetal growth disturbances and other pregnancy complications merits further investigation. PMID- 8423488 TI - Fundal height measurement. Part 3--The effect of maternal position on fundal height measurements. AB - The effect of maternal position on fundal height measurements was studied in 192 nonobese women between 21 and 36 weeks of gestation. Four clinicians participated in the study and each clinician obtained measurements on 48 women. Fundal height measurements were obtained in each of four positions: supine; trunk elevation; knee flexion; and trunk elevation with knee flexion. The sequence in which measurements were obtained was assigned randomly. Clinicians were blinded to the results of their measurements. Measurements obtained in the supine position were largest, and measurements obtained in the trunk elevation with knee flexion position were smallest. Repeated measures analysis of variance demonstrated that measurements obtained in the four positions were significantly different (F = 87.71, df = 3,573, P < .001). A posteriori comparisons demonstrated that measurements obtained in each position were significantly different except for measurements obtained in the trunk elevation and knee flexion positions. These findings indicate that clinicians should be consistent when they position patients to obtain fundal height measurements. PMID- 8423489 TI - Comparison of rural and urban certified nurse-midwives in Arizona. AB - A survey of certified nurse-midwives (CNMs) in Arizona was carried out in 1990 to provide data for maternity service planning in the state. Information was gathered on location and scope of CNM practice, barriers to practice, and the contribution of CNMs to maternity care. Demographic and clinical practice characteristics of urban and rural CNMs were also compared. Urban and rural CNMs are significantly different in terms of education (urban CNMs are much more likely to have master's degrees) and number of years since first certification (urban CNMs have been certified significantly longer). Rural midwives are more likely to be under the age of 40. Health services provided by urban and rural CNMs were compared with each other and with national data. Midwives in rural areas of Arizona are more likely to provide comprehensive nurse-midwifery services than are either urban Arizona midwives or U.S. midwives as a whole. Urban and rural CNMs described lack of physician backup as a major barrier to nurse-midwifery practice in rural areas. Lack of hospital privileges was another major obstacle noted by rural nurse-midwives. Arizona CNMs felt they could provide comprehensive, cost-effective maternity services in rural areas that would improve access to care, patient satisfaction, and maternal and child health outcomes. PMID- 8423490 TI - Findings of the 1991 Annual American College of Nurse-Midwives Membership Survey. PMID- 8423491 TI - Idiosyncratic supraventricular tachycardia after epidural anesthesia. AB - During pregnancy, stress and vigorous exercise often result in pronounced tachycardia. Generally, a vagal stimulatory effort will interrupt the episode; however, intrapartum supraventricular tachycardia may not respond to vagal stimulation, necessitating drug therapy. This article is a case report of idiosyncratic supraventricular tachycardia following initiation of epidural analgesia; use of epinephrine was part of the test dose protocol. Adenosine, chosen for the lack of hypotensive effect associated with verapamil, was administered intravenously with immediate results. Fetal monitoring via scalp electrode provided evidence of fetal well-being during and after the episode. PMID- 8423492 TI - Development and translation of an English-Spanish dual-language instrument addressing access to prenatal care for the border-dwelling Hispanic women of San Diego County. AB - The health of populations residing on either side of the United States-Mexican border has public policy implications for both border communities. A study of barriers to prenatal care for residents of the San Diego, California-Tijuana, Mexico region was conducted. The study was designed specifically to address the availability of health services for women who crossed the border in either direction. This article reports the methodology used in the construction and validation of a dual-language (as opposed to a Spanish language form) survey instrument that was developed for the study. Particular attention is given to the methodology used for language translation to ensure that it would reflect the idiomatic diversity of the multicultural population. PMID- 8423493 TI - Fundal height measurement. Part 1--Techniques for measuring fundal height. AB - Several techniques of measuring fundal height have been described in the literature. Differences between the measurement techniques include the following: whether a tape measure or caliper is used to obtain the measurement; whether the uppermost border of the uterine fundus or the fetal pole in the fundus should be used as the superior landmark; which point on the symphysis pubis should be used as the inferior landmark; whether the tape measure should be held in contact with the skin of the maternal abdomen throughout the length of the measurement; and whether the measurement is obtained in the midline of the maternal abdomen or at the highest point of the uterine fundus or fetal pole. Preliminary studies indicate that the various measurement techniques yield different results. However, the differences between the measurement techniques have not been studied adequately and the superiority of one measurement technique has not been demonstrated. Because the various measurement techniques yield different results, it is important that clinicians be consistent in their measurement technique. Future studies should evaluate and compare the various measurement techniques to determine if one technique yields measurements that are more reliable and valid than other techniques. PMID- 8423494 TI - Evaluation of research studies. Part III: Statistical significance testing. AB - Reading and interpreting research studies is an important and necessary activity of clinical practitioners. Correct evaluation of research findings requires appreciation of certain fundamentals of research methodology and statistics. This article, the third of the series, will review issues pertinent to statistical significance testing. PMID- 8423495 TI - Nurse-midwife. PMID- 8423496 TI - Nurse-midwife. PMID- 8423497 TI - Unqualified specialists in occupational medicine. PMID- 8423498 TI - Risk characterization, risk communication, and risk management. The role of the occupational and environmental medicine physician. AB - Physicians who are board-certified in occupational and environmental medicine often have formal training and experience in toxicology, epidemiology, and industrial hygiene, as well as clinical medicine, human physiology, and pathology, and are well suited to be team members in the various phases of risk assessment, communication, and management. Those with several years of experience in the clinical, as well as administrative practice of occupational and environmental medicine, are engaged in the practical application of risk characterization, risk communication, and risk management on an almost daily basis. We propose that the occupational and environmental medicine physician is in a unique position to bridge the communication gap between those professionals who provide various components of the risk assessment process and those who manage those risks in society. PMID- 8423499 TI - Risk communication and occupational medicine. PMID- 8423500 TI - Health risks and health insurance claims costs. Results for health hazard appraisal responders and nonresponders. AB - The health risks, health perceptions, and health claim costs of health hazard appraisal (HHA) responders and nonresponders were compared in a sample of employees of the Adolph Coors brewery. HHA responders had lower levels of risks than nonresponders. Responders also rated themselves in better health than nonresponders. Despite the health advantage, HHA responders were more likely to file health claims in 1989 than nonresponders and also had significantly greater claims costs. Comparing the top 10% most expensive employees in each group, however, nonresponders had greater claims costs than responders. Distribution patterns also differed based on perceived health status. HHA responders who perceived themselves in poor/fair health status tended to cost less than nonresponders of similar health. Responders perceiving themselves in good/excellent health status cost more than nonresponders. The findings support the "worried well" syndrome in healthy HHA responders. PMID- 8423501 TI - Inherited factors and environmental exposures in cancer risk. AB - Carcinogenesis is a multistage process that results from the interaction of carcinogenic exposures, cellular macromolecules (eg, DNA), and endogenous mutational mechanisms. Involved in these processes are metabolic activation and detoxification of chemical carcinogens, genetic sequences of protooncogenes and tumor suppressor genes, and DNA repair, among others. Each of these vary widely among individuals and can be associated with increased cancer risk. Cytochrome P4501A1, P4502E1 and N-acetyl transferase 2 are examples of enzymes involved in the metabolic activation of potential environmental carcinogens such as polycyclic aromatic hydrocarbons, benzene, and aromatic amines, respectively. Germ-line mutations in these genes are common and associated with abnormal enzymatic function that are mechanistically related to quantitative changes in binding of carcinogens to DNA. Allelic frequencies for these mutations vary among different racial and ethnic populations and may explain, in part, differences in cancer rates. Risk assessments attempt to predict cancer rates in humans using mathematical models that are often based upon limited experimental data. They do not generally incorporate the numerous stages of carcinogenesis or interindividual variation. Thus, sensitive and resistant populations are not sufficiently considered. This limits the accuracy of currently applied risk assessment models. PMID- 8423502 TI - OSHA regulation of ergonomic health. AB - The occupational illnesses most frequently reported to the Occupational Safety and Health Administration (OSHA) are "disorders associated with repeated trauma," a category including noise-induced hearing loss and illnesses caused by ergonomic exposure. In response, OSHA has begun "rule-making" for a standard to control ergonomic exposures. This paper defines ergonomic illness as any diagnosis of gradual onset for which ergonomic exposure increases relative risk. The "Red Meat Guidelines," the model for OSHA's proposed "Ergonomic Safety and Health Program standard," are critically reviewed in terms of basic clinical and epidemiologic concepts. The OSHA 200 log is a cohort study which reaches its entire target population. The use of selected case definitions with the OSHA 200 log is recommended as a powerful, comparable, and cost-effective epidemiologic basis for ergonomic regulation. PMID- 8423503 TI - Occupational exposure of animal caretakers to cyclophosphamide. AB - Little is known about the exposure of animal caretakers to toxic agents during the administration of such chemicals to laboratory animals. In this study, we have investigated the environmental contamination with cyclophosphamide (CP) in an animal laboratory where mice were housed and injected with this compound. Also the contamination of gloves, sleeve protectors, and masks used for personal protection was studied. The uptake of CP by the animal caretakers was determined by the analysis of unmetabolized CP in urine. For the estimation of CP in the air, air samples were taken and filters of the air-circulation system were analyzed. On the filters, amounts of CP were detected corresponding with < 0.1 1.0 microgram/day. Environmental contamination was also measured by analysis of wipe samples taken from different spots (objects and surfaces). The presence of CP was not only observed in the room where the mice were housed and treated with CP but also in adjacent rooms (< 0.02-44 ng/cm2). The gloves used during the injection of CP were always contaminated (2-199 micrograms/pair). No penetration of the gloves was established. The sleeve protectors were incidentally contaminated (< 0.3-10 micrograms) and on the masks no CP was found (< 0.2 microgram). Eighty seven urine samples from four animal caretakers were analyzed for unmetabolized CP. In one sample, CP was detected (0.7 microgram). The results show that in this particular study animal caretakers are exposed to CP during their work. PMID- 8423504 TI - Mental health problems in occupational health care. A pilot study. AB - Purposes of the present study were to report prevalence data of and background factors to mental health problems in Swedish state employees attending occupational health care as compared to primary care patients. The Hopkins Symptom Check List (HSCL-25) was administered to a sample of 39 patients. For comparative reasons, 39 patients matched for sex and age were selected from a primary care sample of 388 patients. The frequency of mental health problems was 33% in occupational health care and 26% in primary care. Patients with mental health problems in both samples presented a diffuse psychosomatic clinical picture and reported a higher degree of loneliness. State employees with mental health problems reported a higher frequency of work-related problems. PMID- 8423505 TI - Exposure of hospital pharmacists and nurses to antineoplastic agents. AB - Adverse effects of antineoplastic drug exposure have been well documented in therapeutically treated patients and proposed as a potential hazard for occupationally exposed populations. Concern stems from accrued evidence of mutagenicity, carcinogenicity, and teratogenicity of many of these drugs. While the clinical significance of occupational exposure is controversial, OSHA issued handling guidelines for these agents in 1986, principally suggesting usage of a vertical laminar airflow biological safety cabinet and good work practices. Industrial hygiene measures of workplace antineoplastic exposure have been few and are limited to air sampling, addressing only the inhalation exposure route. We report here air and surface cyclophosphamide sampling results in a hospital oncology pharmacy and outpatient clinic where OSHA guidelines are in place. Results reveal rare air samples with detectable cyclophosphamide, but multiple surface wipe samples with measurable cyclophosphamide concentrations. Occupational health personnel, therefore, must consider work practices as determinants of surface contamination and recognize the potential importance of dermal and ingestion routes of exposure. PMID- 8423506 TI - A cholinesterase testing program for pesticide applicators. AB - Illnesses associated with the increased use of cholinesterase-inhibiting pesticides have brought about recommendations for monitoring pesticide applicators. The State of California requires medical supervision; however, the supervision and monitoring guidelines are often incorrectly followed. In this retrospective cohort study, 103 worker-years of cholinesterase monitoring are reported. Twenty-four (24%) workers were temporarily removed from spraying (five were removed twice) because their cholinesterase plasma activity levels were below 60% of baseline. Five workers (5%) had mild symptoms of toxicity but none reported a specific incident of exposure. Hispanic workers had fewer significant drops in plasma activity levels and fewer toxic symptoms than white workers. The relative risk of pesticide poisoning was increased in workers whose initial baseline plasma levels were low or if their levels had already dropped to 60%-80% of their baseline previously in the season. Case studies and differences in baselines by month of determination suggest poor monitoring compliance by the companies and employees. Suggestions of how the physician can overcome these problems and improvements of the guidelines are discussed. PMID- 8423507 TI - Preemployment urine substance screening. One-year experience of a large hospital system. AB - Alcohol and drug abuse is estimated to cost our nation more than $160 billion per year in lost productivity, health care costs and crime. Preemployment screening for substances of abuse has become a standard in some industries, including the health care industry. Samaritan Health Services of Phoenix, Arizona, a large health care provider (11,244 employees) began preemployment drug screening and for-cause substance abuse testing March 1, 1991. This paper presents 1-year's statistics for that provider, including problems peculiar to the health care industry, eg, forensic testing stipulation agreements from professional boards. Problems are addressed including the ease of obtaining prescription medications by health care workers ("curbstone consultations") and dealing with a professional population knowledgeable about prescription drug use. It was this corporation's experience that preemployment drug screening can be performed without great hardship to a large health care provider. PMID- 8423508 TI - Preemployment drug screening: the epidemiologic issues. PMID- 8423509 TI - Cancer mortality among workers in the salt-chemical industry. PMID- 8423510 TI - Sixty years after the discovery of pure carcinogens of the polycyclic aromatic class: a tribute to the contributions of Sir Ernest Kennaway to cancer epidemiology and fundamental research in the field of carcinogenesis. PMID- 8423511 TI - Physeal replacement with cultured chondrocytes of varying developmental time: failure to reconstruct a functional or structural physis. AB - Reconstruction of physeal regions excised from the distal femoral chondroepiphysis was attempted in a murine model. Cultured chondrocytes of varying developmental time from the same inbred strain of mice were used for replacement. Vascularity, matrix formation, and cell division, as well as growth, were assessed. The cultured chondrocytes did not produce growth. Consistent with this, cell division, as assessed with incorporation of tritiated thymidine, was not normal. However, the cultured chondrocytes did receive a nutritional supply from the host and did continue matrix formation after transplantation. PMID- 8423512 TI - Double-level cauda equina compression: an experimental study with continuous monitoring of intraneural blood flow in the porcine cauda equina. AB - Compression of the spinal nerve roots may occur clinically at multiple levels at the same time; however, the basic pathophysiology of multi-level compression is largely unknown. In this study, the intraneural blood flow was analyzed continuously in the uncompressed segment between two compression balloons, with a pig used as an experimental model and a thermal diffusion method. At 10 mm Hg compression, there was a 64% reduction of total blood flow in the uncompressed segment compared with pre-compression values. Total ischemia occurred at pressures 10-20 mm Hg below the mean arterial blood pressure. After two-level compression at 200 mm Hg for 10 min, there was a gradual recovery of the intraneural blood flow towards the baseline. Recovery was less rapid and less complete after 2 h of compression. Double-level compression of the cauda equina can thus induce impairment of blood flow, not only at the compression sites, but also in the intermediate nerve segments located between two compression sites, even at very low pressures. These findings may have clinical importance in the understanding of the pathophysiology of multiple-level cauda equina compression. PMID- 8423513 TI - Intraosseous infusion of prostaglandin E2 in the caprine tibia. AB - We evaluated the effects of intraosseously administered prostaglandin E2 (PGE2) within the proximal metaphysis of the goat (caprine) tibia under intraosseous normotensive and hypertensive conditions. PGE2 was administered at 0.5 or 1.0 mg (1 ml vol) twice daily for 10 days via an Osteoport which had been surgically implanted within the proximal tibial metaphysis. Intraosseous hypertension was produced when venous outflow obstruction (VOO) was created by ligation of the popliteal vein, which drained the proximal tibia, and occlusion of the diaphyseal medullary space distally with bone cement. After VOO, the intraosseous pressure measured at the metaphysis increased significantly (p < 0.05) from a baseline mean of 14.9 +/- 4.2 mm Hg to 28.6 +/- 5.3 mm Hg. Serum radioimmunoassays indicated that VOO prolonged the venous drainage of PGE2 from the tibia after an infusion. Static histomorphometric analysis indicated a marked dose-dependent increase in new bone formation in all PGE2 groups at 30 days after the PGE2 infusion. Significant (p < 0.05) formation of new bone occurred, primarily at the subperiosteal and endocortical surfaces, and moderately increased the marrow cavity of cancellous new bone as compared with the VOO-only group and the controls. Bone remodeling indices were also increased by PGE2. The PGE2 infusion, combined with VOO, produced significantly more new bone formation than the PGE2 infusion alone. Intensive marrow fibrosis was associated with the active bone remodeling. PMID- 8423514 TI - Differentiating and antitumor activities of 1 alpha,25-dihydroxyvitamin D3 in vitro and 1 alpha-hydroxyvitamin D3 in vivo on human osteosarcoma. AB - The differentiating and antitumor activities of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) in vitro and 1 alpha-hydroxyvitamin D3 (1 alpha(OH)D3) in vivo were studied with a human osteosarcoma cell line (OST strain). Anti-tumor activity was estimated with the use of 3-(4,5-dimethylthiazol)-2, 5 diphenyltetrazolium bromide (MTT) assay, colony-forming assay, and athymic mouse assay. The intracellular alkaline phosphatase (ALP) activity of tumor cells and production of bone Gla protein (BGP) in culture media were measured to mark osteoblastic differentiation. In addition, the combination of 1 alpha,25(OH)2D3 and cis-dichlorodiammineplatinum(II) (CDDP) was tested by the colony-forming assay and the measurement of ALP activity and BGP production for differentiating and antitumor effects. The assays revealed that 1 alpha,25(OH)2D3 exerted a dose related, growth-inhibitory influence. In the colony-forming assay, the 1 alpha,25(OH)2D3-treated colonies were smaller than the untreated colonies. The ALP activity and the BGP production also increased in relation to dose. In the assay in athymic mice, the relative weight of tumors treated with 1 alpha(OH)D3 at 2.5 nmol/kg was significantly smaller than that of the controls, and no side effects were observed in the 1 alpha(OH)D3-treated mice. Marked tumor chondrogenesis was observed in human osteosarcoma treated with 1 alpha(OH)D3 in athymic mice. The combination of 1 alpha,25(OH)2D3 at 10(-8) M and CDDP at 2 micrograms/ml significantly enhanced both the differentiation and the growth inhibition in vitro. Our study apparently is the first demonstration that vitamin D3 metabolites have an antitumor and differentiating effect on human osteosarcoma cells in vitro and in athymic mice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423515 TI - Torque history of electrically stimulated human quadriceps: implications for stimulation therapy. AB - The time course of knee extension torque was measured in human quadriceps muscles during 30 min of transcutaneous neuromuscular electrical stimulation (NMES). Ninety subjects were divided into six experimental groups (n = 15 per group), which received stimulation at one of the following frequency/duty cycle combinations: 10 Hz/50%, 30 Hz/50%, 50 Hz/50%, 10 Hz/70%, 30 Hz/70%, and 50 Hz/70%. Two-way analysis of variance revealed that the magnitude of the relative torque decrease (the percentage of decrease in torque relative to the initial value) was significantly different between frequencies (p < 0.005) and duty cycles (p < 0.02), with no significant interaction (p > 0.6). Increasing either frequency or duty cycle caused a greater decrease in torque. In spite of this result, there was no significant difference between groups in the total activity (torque-time integral) achieved during the 30 min treatment session. The magnitude of this activity corresponded to only about 7-14 maximum voluntary contractions. Finally, the average torque during the treatment session was significantly different among groups (p < 0.001), being greatest for the 50 Hz/50% group and least for the 10 Hz/70% group. Taken together, these data suggest that a smaller number of longer duration contractions produces the greatest muscle tension. They also suggest that the absolute torque levels achieved with NMES are relatively low compared with voluntary muscular activity. PMID- 8423516 TI - Early postoperative fixation of tibial components: an in vivo roentgen stereophotogrammetric analysis. AB - The fixation of cemented and noncemented tibial components in 19 total knee arthroplasties was examined 3 to 10 weeks postoperatively with roentgen stereophotogrammetric analysis. Physiologic external forces were applied either in outward-inward rotation or as eccentric posterior loading generated by squatting. In one case there was no motion, while in the remaining 18 cases relative interface motion of up to 1.3 mm was found. In some cases, the largest motion was found during inward-outward rotatory stress, while in others, the largest motion was found during squatting. Posterior tilt was weakly correlated with posterior displacement of the femoral component on the tibia during squatting (r2 = 0.323, p < 0.05). For the noncemented cases, the initial fixation to bone was probably insufficient, and ingrowth of bone would not have been achieved. For the cemented cases, motion of the implant was reduced. Given the short postoperative time and the probable absence of any substantial fibrous tissue membrane, we suggest that the observed motion represented elastic deformation of the bone. PMID- 8423517 TI - Maintenance of set force in anterior cruciate ligament grafts. AB - This study was undertaken to determine how accurately total graft force and load sharing between graft segments could be set and maintained during augmented anterior cruciate ligament (ACL) reconstruction in the goat knee. Special procedures were developed to reduce the effect of tissue creep and to overcome difficulties in the setting of graft force. Five knees from goat cadavers were reconstructed using a bone-tendon-bone graft (PT) and a synthetic augmentation device (LAD). Prescribed levels of total graft force and load-sharing between the autograft and LAD were set under a standardized external joint load. Immediately after fixation, the set force declined an average of 9 and 3% in the LAD and PT, respectively. After three subsequent exercise sequences, the set forces fell from their initial level by an average of 25% for the LAD and 28% for the PT. An analysis of variance did not show the loss of force with exercise to be statistically significant. We conclude from this in vitro study that our method can be used to set forces in an ACL reconstruction with reasonable maintenance of load-sharing but that losses of approximately 30% of total graft force after exercise of the reconstructed joint are to be expected. PMID- 8423518 TI - Evaluation of a local microsphere injection method for measurement of blood flow in the rabbit lower extremity. AB - The precision of a modified microsphere technique to measure blood flow in the hind limb of the rabbit was determined. Regional (local) injections made via a catheter in the aorta and blood withdrawn through a catheter in the hind limb enabled deposition of a high concentration of microspheres in the tissues while minimizing the number of microspheres needed for accurate measurement. 95Niobium and 141cerium-labeled 16 microns diameter microspheres were used. Blood flow was measured in 15 locations of muscle and skin for simultaneous injections into the aorta and left ventricle (group A), simultaneous injections of a mixture into the left ventricle (group B), and ventricular injections given 1 h apart (group C). No significant differences in mean blood flow measurements were found. Correlation coefficients between local and ventricular injections ranged from 0.89-0.99 and between 0.994 and 0.999 for two simultaneous ventricular injections. The variability over time was greater, with correlations between the first and second injections ranging from 0.13-0.939. Precision of the local technique compared with the ventricular technique can be kept within 30% with five or six animals. The advantage of this technique over ventricular injections is in the measurement of blood flow in small tissue samples when the number of microspheres deposited from ventricular injections is insufficient. PMID- 8423519 TI - Blood flow changes in the tibia during external loading. AB - The influence of external load on the blood flow of the rabbit tibia was assessed by laser Doppler flowmetry (LDF). Blood flow readings were obtained from three adolescent and three adult female New Zealand White rabbits whose tibiae previously had been pinned with modified orthopaedic pins. Readings were obtained from the exposed mid-shaft of the tibia of each hind leg before, during, and after loading. The loading was either static or sinusoidal intermittent. The LDF resulted in a reading from the tibia which was consistent with the recordings of blood flow; the traces were similar to the pulsatile nature of pulse pressure recordings. After loading of the tibia, in both the adolescent and adult rabbits the effects were an increase in blood flow, as measured with LDF, on the tensile aspect, and a decrease in flow on the compressive side. These changes were statistically significant (p < 0.01) as measured by a Student t test. This change in flow reached a plateau with change in strain. Furthermore, the changes in flow produced by loading continued as long as the load was applied. A response similar to reactive hyperaemia occurred on the compressive aspect of the tibia; there was an increase of blood flow above the resting level after removal of the static load. The effect of a sinusoidal intermittent load was less than that of a static load, and the differences in flow between tensile and compressive aspects was not significant. The changes induced offer further insight into factors controlling bone remodelling. PMID- 8423520 TI - A quantitative study of the routes of blood flow to the tibial diaphysis after an osteotomy. AB - Techniques of tissue exclusion have been used previously in qualitative investigations of the vascularity of long bones, after experimental fracture; we quantified their effects on bone blood flow in rabbits. Thirty-six adult rabbits were divided into three groups in which flow was measured, with the microsphere technique, 1 and 2 weeks after osteotomy. In Group 1, osteotomy of the tibial shaft only was done; in Group 2, osteotomy was done with exclusion of the periosteum and muscle by a silicone rubber sheath; and in Group 3, osteotomy was done with exclusion of the marrow by reaming and insertion of an intramedullary nail. All involved limbs were immobilized in a cast. In Group 1, cortical flow increased but marrow flow did not change, which suggests that the changes in cortical flow were mediated by a supply paralleling that of the marrow. In Group 2, the changes in cortical flow were abolished, which implies that this parallel supply is from the periosteum and extraosseous tissues. In Group 3, cortical flow was not significantly reduced, which demonstrates recruitment of this periosteal and extraosseous supply. These results lend support to the hypothesis that the blood supply to the healing diaphysis is principally from the periosteum and extraosseous tissues during the early peak period of blood flow. PMID- 8423521 TI - The mechanical properties of skeletally mature rabbit anterior cruciate ligament and patellar tendon over a range of strain rates. AB - The effect of strain rate on the mechanical properties of the rabbit anterior cruciate ligament (ACL) and patellar tendon (PT) was evaluated. The medial portion of the ACL was loaded to tensile failure at rates of 0.003, 0.3, and 113 mm/s, and the middle third of the PT was loaded at rates of 0.008, 0.8, and 113 mm/s. The load was recorded with a high-speed measurement plotting system, and each test was videotaped for strain analysis. The nonlinear portion of the stress strain curve was curve-fit to an exponential function having two nonlinear constants, representing the initial modulus and rate of change of the modulus. The modulus of the rabbit PT was found to be 89% higher than that of the ACL. The initial modulus and rate of change of the modulus also were greater for the PT than for the ACL. The modulus of the PT was shown to be more sensitive to strain rate than that of the ACL; a 94% increase was observed for the PT, and a 31% increase was observed for the ACL. There was no effect of strain rate on the mode of failure of either the ACL or the PT; all but three of the specimens failed at the insertion site. PMID- 8423522 TI - Proteoglycans in the compressed region of human tibialis posterior tendon and in ligaments. AB - Proteoglycan content and tissue morphology were examined in tendons and ligaments from 24 cadavers, ranging in age at the time of death from 1.5 months to 83 years. The region of the human tibialis posterior tendon that passes under the medial malleolus was characterized by cells having a rounded shape, positive staining with alcian blue, and higher glycosaminoglycanuronic acid content than in the more proximal region of the same tendon. Analysis of proteoglycans by sodium dodecyl sulfate/polyacrylamide gel electrophoresis indicated that the predominant small proteoglycan of the proximal/tensional region was decorin, whereas two types of small proteoglycans (decorin and biglycan) and large proteoglycans were present in the region passing under the medial malleolus and presumably subjected to compressive and shear forces in addition to tension. The pattern of proteoglycan accumulation in the compressed region of tendon was basically similar for all individuals and showed no distinctive trends related to age after puberty. In terms of type and amount of proteoglycan, the patellar tendon was like the tensional region of the tibialis posterior. Glycosaminoglycan content in the lateral collateral ligament and anterior cruciate ligament, however, was twofold higher than in the tendons. The ligaments contained large as well as small proteoglycans, just as in the compressed region of tendon. PMID- 8423523 TI - Two processes of bone remodeling in plated intact femora: an experimental study in dogs. AB - Bone loss is known to occur under plates used for internal fixation. Its exact location and extent and its relationship to the plate-bone contact area was investigated in 36 adult beagles. Plates with two different contact areas were fixed unilaterally to intact femora for periods of 8 and 24 weeks. After removal, the bones were assessed radiographically, histologically, histomorphometrically, and biomechanically. Two processes that affect the bone after plating became apparent. One is a short-term process, possibly caused by the interference of the plate with the blood supply to the cortex. This is followed by necrosis, which induces porosis. The second independent process is caused by the tendency of the bone to adapt to the new state of mechanical loads that results from the load sharing by the plate. The first stage of this process is expressed through swift changes in the bone cross-sectional area under the plate by remodeling in all three envelopes: haversian, periosteal, and endosteal. This stage is followed by a gradual remodeling towards a steady state, in which the final cross-section of the bone is attained. Both processes affected the bone simultaneously during plate fixation. The process caused by the vascular interference ceases after some time, and only the process due to stress-shielding prevails for a long period. PMID- 8423524 TI - An intracellular calcium release inhibitor TMB-8 suppresses renal nerve stimulation-induced antinatriuresis in dogs. AB - Effects of nifedipine and TMB-8 on antinatriuresis induced by renal nerve stimulation (RNS) were examined in pentobarbital-anesthetized dogs. RNS (1 Hz) decreased urine flow rate, urinary sodium excretion rate and fractional excretion of sodium and increased renal norepinephrine efflux and renal venous plasma renin activity with little changes in renal hemodynamics. Intrarenal arterial infusion of nifedipine (0.1 microgram/kg/min) or TMB-8 (50 and 100 micrograms/kg/min) increased basal urine flow rate, urinary sodium excretion rate and fractional excretion of sodium without affecting renal venous plasma norepinephrine concentration or plasma renin activity. Neither nifedipine nor TMB-8 affected the RNS-induced increases in norepinephrine efflux and plasmin renin activity. The RNS-induced decreases in urinary sodium excretion rate and fractional excretion of sodium were suppressed during the TMB-8 infusion, whereas nifedipine failed to affect these urinary responses. These results raise the possibility that the release of intracellular calcium from TMB-8-sensitive stores, but not the influx of extracellular calcium through dihydropyridine-sensitive calcium channels, participates in neural control of tubular sodium reabsorption in the dog kidney. PMID- 8423525 TI - Postnatal development, sex-related difference and hormonal regulation of acetohexamide reductase activities in rat liver and kidney. AB - A variety of patterns of postnatal development were observed for acetohexamide reductase activities in microsomes and cytosols from the liver and kidney of male and female rats. Furthermore, there were pronounced sex-related differences in the activities of liver and kidney microsomes. Prepubertal and neonatal testectomies decreased markedly the activities in liver and kidney microsomes of adult male rats to the female levels. The activities in liver and kidney microsomes decreased by prepubertal testectomy and the activity in kidney microsomes decreased by neonatal testectomy were restored to control levels by testosterone propionate treatment, whereas the activity in liver microsomes decreased by neonatal testectomy was unaffected by the same treatment. Prepubertal or neonatal testectomy had no effect on the activity in liver or kidney cytosol of adult male rats. Prepubertal ovariectomy decreased the activities in liver and kidney cytosols of adult female rats and the decreased activities were restored to control levels by estradiol benzoate treatment. Based on these results, we propose that the postnatal development and sex-related difference of acetohexamide reductase activities in rat liver and kidney can be regulated by sex hormones such as testosterone propionate and estradiol benzoate. PMID- 8423526 TI - Serotonin-induced contraction in porcine coronary artery: use of ergolines to support vascular 5-hydroxytryptamine2-receptor heterogeneity. AB - Serotonin-induced contraction in porcine coronary artery was studied in ring segments of coronary artery without endothelium. 5-hydroxytryptamine (5-HT), 5 methoxytryptamine (5-MeOT) and alpha-methyl-5-HT (alpha Me-5-HT) concentration dependently contracted vessels with similar maximal force. The agonist rank order potency was 5-HT > alpha Me-5-HT > 5-MeOT. Neither prazosin (1 microM) nor tetrodotoxin (0.3 microM) significantly altered 5-HT-induced contraction, ruling out activation of alpha-1 adrenoceptors and sodium channels in the contractile response, respectively. Using 5-MeOT as the prototype agonist, cyanopindolol blocked contraction with an antagonist dissociation constant lower than expected for 5-HT1-receptor blockade and ICS 205-930 (10 microM) did not affect 5-MeOT induced contraction. Five structurally distinct 5-HT2-receptor antagonists (ketanserin, cisapride, spiperone, MDL11939, ICI169369) blocked 5-HT-induced contraction with antagonist dissociation constants similar to reported 5-HT2 receptor affinities. Two ergoline-based 5-HT2-receptor antagonists, LY53857 and sergolexole, blocked 5-HT-induced contraction with antagonist dissociation constants lower than expected for vascular 5-HT2-receptor blockade. Based on the agonist profile and the fact that ICS 205-930 and cyanopindolol were not potent antagonists, the 5-HT receptor-mediating contraction does not represent either the 5-HT1A/B/C/D, 5-HT3 or 5-HT4 receptor. Rather, based on the affinity of several established 5-HT2-receptor antagonists, a 5-HT2 receptor mediates contraction in porcine coronary artery. However, the low antagonist affinity of the ergolines (i.e., LY53857 and sergolexole) suggests that heterogeneity of 5 HT2 receptors may exist among species. PMID- 8423527 TI - Regio- and stereoselective propranolol metabolism by 15 forms of purified cytochromes P-450 from rat liver. AB - Regio- and stereoselectivity of cytochrome P-450-mediated propranolol metabolism (4-, 5- and 7-hydroxylations and N-desisopropylation) was studied using 15 purified cytochrome P-450 species. With each purified cytochrome P-450 species, the regioselectivity was distinct and different between the two optical isomers used as substrates. The stereoselectivity was different depending on the position of propranolol to be metabolized. The regio- and stereoselectivity was altered when substrate concentration was altered, suggesting that the kinetics of the reactions are different depending on the positions of propranolol to be metabolized. Furthermore, the selectivity and its manner of alterations with substrate concentrations were different among all cytochrome P-450 species used. Propranolol, with its multiple metabolic pathways and optical isomers, is an extremely interesting substrate for characterization of cytochrome P-450 species. PMID- 8423528 TI - Pharmacological profile of NPC 17742 [2R,4R,5S-(2-amino-4,5-(1, 2-cyclohexyl)-7 phosphonoheptanoic acid)], a potent, selective and competitive N-methyl-D aspartate receptor antagonist. AB - 2R,4R,5S-(2-amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoic acid) (NPC 17742), the most potent isomer of the mixture 2-amino-4,5-(1,2-cyclohexyl)-7 phosphonoheptanoic acid (NPC 12626), was evaluated for activity in tests associated with receptors for excitatory amino acids. In receptor binding assays, NPC 17742 was selective for the N-methyl-D-aspartate (NMDA) receptor with a potency comparable to that of D(-, -3-(2-carboxypiperazine-4-yl)propyl-1 phosphonic acid. Like (+/-)cis-4-phosphono-methyl-2-piperidine carboxylic acid (CGS 19755) and (+/-)(E)-2-amino-4-methyl-5-phosphono-3-penteneoic acid (CGP 37849), NPC 17742 competitively inhibited NMDA-induced enhancement of 1-[(2 thienyl)cyclohexyl]piperidine binding to the NMDA receptor ionophore and partially inhibited [3H]glycine binding to strychnine-insensitive sites. In contrast, NPC 17742 and CGP 37849 inhibited Mg(++)-stimulated 1-[(2 thienyl)cyclohexyl]piperidine binding in a noncompetitive fashion. In voltage clamped Xenopus oocytes expressing excitatory amino acid receptors, NPC 17742 (pKB = 6.91) was equipotent with CGP 37849 (pKB = 7.17) in inhibiting NMDA induced inward currents. Likewise, NPC 17742 (ED50 = 2.68 mg/kg) was equipotent with CGP 37849 and CGS 19755 in blocking NMDA-induced convulsions, but was less potent than these two compounds in the maximal electroshock test. Unlike CGP 37849 or CGS 19755, NPC 17742 potently antagonized seizures induced by pentylenetetrazol. In a model of global ischemia, low doses of NPC 17742 given either before or after ischemic result were effective in blocking damage to hippocampal CA1 neurons. The pharmacologic responses to NPC 17742 occurred at doses 30- to 300-fold lower than the acute lethal dose.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423529 TI - Vasomotor responses of canine coronary arterial rings to NG-monomethyl-L-arginine and N omega nitro L-arginine methyl ester. AB - The actions of NG-monomethyl-L-arginine (L-NMMA) and N omega-nitro L-arginine methyl ester (L-NAME) on canine coronary arterial rings were compared with effects on rat aortic rings. After incubation of rat aortic rings with indomethacin (5 x 10(-6) M) and preconstriction with phenylephrine (10(-7) M), L NMMA (2.5 x 10(-4) M) caused an increase in tension when endothelium remained intact (+1.1 +/- 0.2 g). L-NMMA had no effect when endothelium was absent. After incubation of canine coronary arterial rings with indomethacin and preconstriction with prostaglandin F2 alpha (10(-6) M), L-NMMA (2.5 x 10(-4) M) increased tension (+39.9 +/- 7.9% of PGF2 alpha-induced constriction) when endothelium was intact, but L-NMMA caused a significant reduction in tension when endothelium was removed (Emax -52.2 +/- 10.3%; P < .05). The reduction in tension after L-NMMA was greater in the absence of indomethacin (Emax -79.8 +/- 4.1%; P < .05). It has been suggested previously that biotransformation of L-NMMA to L arginine may have contributed to vasorelaxation; L-arginine is the endogenous substrate of nitric oxide synthase. However L-arginine (10(-3) M) did not affect the fall in tension produced by L-NMMA (Emax -69.0 +/- 14.2% in the absence of indomethacin). We also found that incubation with the protein synthesis inhibitor, cycloheximide, did not block the L-NMMA-induced fall in vascular tension; in fact, it increased the magnitude of the relaxant effect (-95.4 +/- 2.5%, experiments performed without indomethacin).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423530 TI - Further evidence that the guinea pig tracheal contractile serotonergic receptor is a 5-hydroxytryptamine2 receptor: use of 5-methyltryptamine and dipropyl-5 carboxamidotryptamine. AB - The guinea pig tracheal contractile serotonergic receptor shows pharmacological similarity to both 5-hydroxytryptamine2 (5-HT2) and 5-hydroxytryptamine1C (5 HT1C) receptors. The present in vitro study utilizes 5-methyltryptamine (5-MT), a high-affinity 5-HT1C receptor ligand, and dipropyl-5-carboxamidotryptamine (DP-5 CT), a low-affinity 5-HT1C receptor ligand, as tools to probe the role of 5-HT1C vs. 5-HT2 receptors in tracheal contractility. Both tryptamines contracted trachea with lower potency (-log EC50 < 5) than 5-HT (-log EC50 = 6.98). Maximum contraction to both 5-MT and DP-5-CT was only 50 to 80% of the maximum response to 5-HT, suggesting that these compounds were partial agonists relative to 5-HT. Guinea pig tracheal contraction to 5-HT, 5-MT and DP-5-CT was inhibited by 1-(1 naphthyl)piperazine (1-NP, 30 nM). The apparent antagonist dissociation constants (KB) for 1-NP were similar when 5-HT (-log KB, K = 8.70 +/- 0.17) or 5-MT (- log KB = 8.40 +/- 0.12) were used as agonists. Although 1-NP also blocked tracheal contraction to DP-5-CT, solubility limitations with DP-5-CT did not permit calculation of a KB value for 1-NP. Nevertheless, these data indicate that 5-MT and DP-5-CT interact with the same receptor as 5-HT. As antagonists, both 5-MT and DP-5-CT inhibited tracheal contraction to 5-HT, indicating interaction with the same receptor as 5-HT and confirming partial serotonergic agonist activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423531 TI - Cholinergic modulation of N-methyl-D-aspartate-evoked [3H]norepinephrine release from rat cortical slices. AB - Previous work has shown that N-methyl-D-aspartate (NMDA) receptor activation inhibits muscarinic receptor-mediated phosphoinositide hydrolysis in brain slices. To further explore the potential interactions between NMDA receptors and cholinergic receptors, the effects of cholinergic agonists and NMDA on [3H]norepinephrine (NE) release from rat cortical slices were determined. Slices were labeled with [3H]NE, washed and treated with various agonists by transferring the slices through a series of vials at 1-min intervals. Radioactivity remaining in the medium was then quantitated to determine the fractional release of [3H]NE from the slices. Carbachol (30-3000 microM) slightly stimulated [3H]NE release from a basal level of 0.10 to approximately 0.35 fractional release by itself and significantly enhanced the effect of 250 microM NMDA (3.6 fractional release for NMDA and 5.3 for carbachol + NMDA) in a concentration-dependent manner. Carbachol (1 mM) increased the maximal response but had no effect on the EC50 of NMDA. Atropine (1 microM) significantly attenuated the effect of carbachol alone and the potentiation of NMDA-evoked [3H]NE release by carbachol, whereas d-tubocurarine (10 microM) inhibited the effect of carbachol alone but had no effect on the enhancement of the NMDA response by carbachol. Mecamylamine (100 microM) inhibited the effect of carbachol alone, but also inhibited the NMDA-evoked response with an IC50 of 16 microM. The nicotinic agonist, dimethylphenylpiperazinium (DMPP) stimulated [3H]NE release (approximately 0.4 fractional release at 30 microM) and also potentiated NMDA-stimulated [3H]NE release (2.0 above NMDA alone). d Tubocurarine, but not atropine, partially inhibited DMPP-stimulated [3H]NE release, but neither antagonist altered the enhancement of NMDA-stimulated [3H]NE release by DMPP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423532 TI - Chronic cocaine disruption of estrous cyclicity in the rat: dose-dependent effects. AB - The effects of cocaine on cyclic reproductive function in females remain largely unknown. In this study, we sought to define the range of doses of cocaine effective in disrupting estrous cyclicity and inhibiting ovulation. Estrous cyclicity was monitored daily by vaginal cytology. Group 1 consisted of rats receiving no treatment. Group 2 consisted of rats injected daily with saline s.c. Groups 3 to 6 consisted of rats injected with 1, 5, 10 and 20 mg/kg/day of cocaine HCl s.c., respectively. Group 7 consisted of rats that were food restricted to allow weight gains comparable to those of group 6. Our results indicate a dose-dependent effect of cocaine on estrous cyclicity with an estimated IC50 of 8.5 mg/kg/day (i.e., the dose of cocaine required to inhibit the number of proestrus: estrus events per 3-week period of analysis by 50%). Over 50% of the rats with cycle disruption on 10 mg/kg/day of cocaine, but almost none of those with cycle disruption on 20 mg/kg/day, returned to normal cyclic patterns after cessation of cocaine treatment. Serum luteinizing hormone levels were reduced 53 and 74% by 10 and 20 mg/kg/day of cocaine, respectively, with an IC50 of 8.9 mg/kg/day. In contrast, cocaine had no significant effect at any of the tested dosages on serum follicle-stimulating hormone or prolactin levels. Ovulation rates were significantly reduced at both 10 and 20 mg/kg/day of cocaine (IC50 = 11 mg/kg/day of cocaine).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423533 TI - Identification of an orally active, nonpeptidyl oxytocin antagonist. AB - L-366,509, a member of a novel class of nonpeptidyl compounds, has been characterized as an orally active oxytocin (OT) antagonist. L-366,509 exhibits a moderate binding affinity (K(i) values, 370-780 nM) for the rat, rhesus and human uterine OT receptor. L-366,509 also binds to vasopressin receptor subtypes (arginine vasopressin-V1 and V2) with measurable affinity in rat (K(i) values, 25 30 microM) and primate (K(i) values, 2-6 microM) tissues. In rat uterine slices, L-366,509 inhibits (IC50 = 1.6 microM) the stimulation of phosphatidylinositol turnover induced by OT but not bradykinin. In the rat isolated uterus, L-366,509 is a competitive and reversible OT antagonist (pA2 = 7.32). In vivo, L-366,509 given i.v. (10 mg/kg) or intraduodenally (10-50 mg/kg) to rats causes a marked and long-lasting inhibition of OT-stimulated uterine activity. OT antagonist activity in a pregnant rhesus macaque (approximately day 135 gestation) is also observed with L-366,509 after i.v. or p.o. dosing. L-366,509 represents a prototype for a new chemical class of OT antagonists with significant p.o. bioavailability. PMID- 8423534 TI - Pharmacokinetic evaluation of drug interactions with anti-human immunodeficiency virus drugs. VI. Effect of the calcium channel blocker nimodipine on zidovudine kinetics in monkeys. AB - Coadministration of zidovudine (AZT) and nimodipine, a calcium-channel blocker, is a potential therapeutic regimen in acquired immunodeficiency syndrome patients based on the report that nimodipine can prevent human immunodeficiency virus induced neurotoxicity in vitro. An evaluation of the pharmacokinetics of AZT and its glucuronide metabolite 3'-azido-3'-deoxy-5'-O-beta-D glucopyranurosylthymidine (GAZT) in the presence and absence of nimodipine in monkeys was undertaken. After 20 mg/kg of AZT given i.v. in the presence and absence of nimodipine, nimodipine caused a significant reduction (41%) in the volume of distribution of AZT at steady state and a 22% decrease in total clearance. The disposition of GAZT was also influenced by nimodipine, causing a large increase in its area under the plasma concentration-time curve. Renal excretion data for AZT and GAZT, although inconclusive, suggested nimodipine caused a decrease in the renal clearance of AZT with a minimal change in the renal clearance of GAZT. The combined effects of nimodipine on the clearance of AZT and volume of distribution at steady state produced no change in its elimination half-life. PMID- 8423535 TI - Alterations by a thromboxane A2 analog (U46619) of calcium dynamics in isolated rat cardiomyocytes. AB - The mechanism by which thromboxane A2 (TXA2) causes its detrimental actions on the myocardium during ischemia and reperfusion injury is unknown. The present study was designed to investigate the influence of U46619, a stable TXA2 analog, on intracellular Ca transients in electrically stimulated single neonatal rat ventricular myocytes by using spectrofluorometric analysis of fura-2-Ca binding. Administration of U46619 increased basal and peak Ca concentrations as well as width of electrically induced Ca transients in a concentration-dependent manner (0.1-1 microM) during a 1-hr exposure. Exposure to 10 microM U46619 caused irregular Ca transients and a marked increase in cytosolic-free Ca concentration. The effects of U46619 were antagonized by the TXA2 receptor antagonist SK&F95585 (2 microM), dibutyryl cyclic AMP (1 mM), verapamil (1 microM) and ryanodine (1 microM). U46619 did not affect the increase in cytosolic Ca induced by KCl (90 mM) depolarization. Caffeine (10 mM)-induced Ca release from the sarcoplasmic reticulum was enhanced markedly in U46619-treated cells. Significant lactate dehydrogenase leakage from the myocytes did not occur at 1 to 10 microM U46619. These results indicate that the increase in Ca transients by U46619 is a receptor mediated process leading to a Ca accumulation in the sarcoplasmic reticulum which is likely to be responsible for an enhanced cytosolic Ca during excitation contraction coupling. Thus, the identification of U46619-induced alterations of Ca dynamics appears to provide, at the cellular level, a direct role for TXA2 during myocardial ischemia and reperfusion. PMID- 8423536 TI - Rat kidney endothelin receptors in ischemia-induced acute renal failure. AB - The current study was designed to assess the possible changes in kidney endothelin (ET) receptors in a model of ischemia-induced acute renal failure. In unilaterally nephrectomized (right kidney) Sprague-Dawley rats, the left renal artery was occluded for 30 min under pentobarbital anesthesia (40 mg/kg i.p.). Body temperature was maintained at 37 degrees C. Sham-operated rats were used as control. Plasma creatinine levels were measured and ET receptors were quantitated by binding of [125I]ET-1 to membranes prepared from the renal cortex at 0, 2, 5 and 24 hr after reperfusion. There was a time-dependent increase in plasma creatinine levels as well as in [125I]ET-1 binding to cortical membranes at 2, 5 and 24 hr postreperfusion. Saturation binding experiments using [125I]ET-1 and [125I]ET-3 indicated that this increase in binding was due to an increase in affinity without significant change in maximum binding. The affinities were 219, 134, 100, 69 and 80 pM for [125I]ET-1 and 320, 273, 130, 168 and 80 pM for [125I]ET-3 and, for sham, 0, 2, 5 and 24 hr postreperfusion, respectively. These data support the hypothesis of ET involvement in the pathophysiology of acute renal failure in rats. PMID- 8423537 TI - Effect of Ca++ channel blockers on energy level and stimulated insulin secretion in isolated rat islets of Langerhans. AB - To investigate the effect of calcium channel blockade on intracellular energy levels and stimulated insulin secretion in isolated rat pancreatic islets, five different blockers of calcium channels were used. Insulin secretion stimulated with 16 mM glucose, 40 mM KCl or 20 mM alpha-ketoisocaproic acid was inhibited dose dependently by nifedipine, diltiazem, flunarizine and verapamil, albeit with different potencies. Nifedipine and flunarizine were more potent than diltiazem and verapamil. omega-Conotoxin GVIA (100 nM) had no significant effect on insulin release with all stimuli tested, although it caused approximately 20% inhibition of the late phase of secretion stimulated with high glucose. The doses of L-type channel antagonists and of flunarizine chosen for the measurements of cellular energy levels gave 60 to 80% inhibition of total stimulated insulin release. The [ATP]/[ADP] ratio, with 5 mM glucose in the perifusion medium, was greater when these channel blockers were present than in controls, whereas it was smaller with omega-conotoxin. The rises in the nucleotide ratio elicited by 16 mM glucose were not affected by any of the antagonists tested. Thus, influx of Ca++ and a consequent rise in its intracellular level are unlikely to be the primary causal event in stimulation of energy synthesis which occurs upon addition of high concentrations of metabolic secretagogues. PMID- 8423538 TI - A novel-response procedure enhances the selectivity and sensitivity of a triazolam discrimination in humans. AB - Placebo-appropriate responding in drug discrimination can be difficult to interpret because such responding can indicate either the absence of any drug effect or the absence of a specific drug effect. This study addressed the overinclusiveness of placebo-appropriate responding by providing a response alternative for novel-drug effects (i.e., effects unlike the training stimuli). This "novel-response procedure" used instructions that indicated that only responses on a novel-appropriate manipulandum would be reinforced in the presence of novel drug effects. Four healthy male volunteers (ages 19-32) were trained to discriminate 0.32 mg/70 kg of triazolam from placebo. Then, dose-effect curves were determined for triazolam (0.1-0.32 mg/70 kg) and d-amphetamine (5 and 20 mg/70 kg) with a standard two-response procedure (drug vs. placebo) and the novel response procedure. Triazolam produced dose-related increases in triazolam appropriate responding with both procedures. d-Amphetamine produced predominantly placebo-appropriate responding with the two-response procedure and predominantly novel-appropriate responding with the novel-response procedure. Unexpectedly, the triazolam dose-effect curve obtained with the novel-response procedure was shifted to the left relative to the two-response procedure for discrimination measures. A similar effect was evident for both the triazolam and d-amphetamine dose-effect curves for some self-report measures. Because of the increased selectivity of placebo-appropriate responding and the increased potency of the drug stimulus, the novel-response procedure may represent a methodological advance for drug discrimination research. PMID- 8423539 TI - Stimulation of endogenous prostacyclin protects the reperfused pig myocardium from ischemic injury. AB - Several attempts have been undertaken to reduce the severity of ischemic myocardial injury by exogenous administration of eicosanoids and by modification of endogenous eicosanoid production. The present study investigates whether defibrotide, a compound that stimulates endogenous prostacyclin (PGI2), has a beneficial effect in experimental ischemic myocardial injury. Anesthetized, open chest minipigs were subjected to 1 h of coronary artery occlusion, followed by 3 h of reperfusion. Defibrotide (32 mg/kg x h) or its vehicle were infused i.v. throughout the experiment. Defibrotide increased cardiac PGI2 formation 3- to 4 fold greater than control (P < .05). Thromboxane levels remained unchanged. Irreversible ischemic injury, as identified by negative tetrazolium staining, amounted to 44 +/- 6% of the area at risk in pigs receiving vehicle but was reduced to 23 +/- 4% by defibrotide (P < .05). This reduced tissue injury in defibrotide-treated pigs was associated with improved functional recovery (left ventricular pressure, + dP/dtmax), during early reperfusion. Recovery did not occur in vehicle-treated pigs. Collagen (2 micrograms/ml)-induced platelet aggregation ex vivo was increased in vehicle-treated pigs during ischemia and reperfusion, but not in animals treated with defibrotide. Polymorphonuclear neutrophil leukocyte accumulation in the ischemic border zone was reduced from 59 +/- 17 cells/mm2 in vehicle-treated pigs to 17 +/- 9 cells/mm2 by defibrotide (P < .05). Pretreatment of the animals with indomethacin (3 mg/kg) prevented the reduction of infarct size and polymorphonuclear neutrophil leukocyte infiltration by defibrotide. Indomethacin increased infarct size in vehicle- and defibrotide treated pigs by 71 and 59%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423540 TI - Detection and mechanisms of formation of S-(6-purinyl)glutathione and 6 mercaptopurine in rats given 6-chloropurine. AB - 6-Chloropurine (CP) has antitumor activity against animal and human neoplasms, but the mechanism is unclear. Recently, we have shown that S-(6 purinyl)glutathione (PG), a putative metabolite of CP, is metabolized in vivo to yield the antitumor drug, 6-mercaptopurine (6-MP). In this study, CP metabolism to PG and 6-MP was investigated in an effort to provide further insights into the mechanism of CP antitumor activity. Rat hepatic and renal glutathione S transferases metabolized CP to PG; Vmax values for liver and kidney cytosol were 166 and 24 nmol/mg of protein/min, respectively. PG was isolated and characterized by fast atom bombardment mass spectrometry from the bile of rats given CP. When rats were given CP (14 mumol/kg), PG excretion was linear with time for up to 1 hr; nearly 80% of the PG excreted at 2 hr was excreted at 1 hr. Rats given CP (10-1200 mumol/kg) excreted at 1 hr into bile nearly 18% of the dose as PG; rats given CP (400-1200 mumol/kg) excreted at 24 hr into urine nearly 4% of the dose as 6-MP and its further metabolites, 6-methylthiopurine and 6 thiouric acid. CP, PG, 6-MP, 6-methylthiopurine and 6-thiouric acid were also detected in plasma, liver and kidney of rats given CP (1200 mumol/kg); in these tissues, maximum CP concentrations were observed at 30 min, as compared to 60 to 180 min, and plasma CP concentrations were higher than those detected in liver or kidney. Liver or kidney CP metabolite concentrations at 30 to 120 min were, however, higher than those detected in plasma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423541 TI - Autoradiographic mapping of M3 muscarinic receptors in the rat brain. AB - The regional distribution of putative M3 receptors was studied by quantitative autoradiography of rat brain sections labeled with 4-diphenylacetoxy-N-methyl piperidine methioxide. The radioligand appears to label multiple muscarinic receptor subtypes in direct binding assays, as evidenced by competition experiments with unlabeled pirenzepine and 11-([2-[(2-(diethylamino)methyl]-1- piperidinyl]acetyl-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodi azepine-6-on. Conditions favoring the selective labeling of M3 receptors were determined on the basis of the competition studies and were subsequently applied to regional mapping of M3 receptor binding. Under the conditions used, autoradiographic images may be obtained with both 9-fold M3-to-M1 selectivity and 12-fold M3-to-M2 selectivity. Although distributed widely at relatively low prevalence throughout the neuraxis, M3 receptors are apparently enriched relative to M1 and M2 receptors in several diencephalic and brain stem regions including thalamic and hypothalamic nuclei, the substantia nigra, superior colliculus, periaqueductal gray and the pons. The present results suggest that the binding of 4 diphenylacetoxy-N-methyl-piperidine methioxide, under appropriate conditions, may permit direct estimation of M3 receptors in mammalian brain. PMID- 8423542 TI - Autoradiographic localization of dopamine D1 receptors in the human kidney. AB - The pharmacological profile and the anatomical localization of the selective D1 receptor antagonist [3H][R]-(+)-8-chloro-2,3,4,5-tetrahydro-5-phenyl-1H-3 benzazepin-7 -al- hemimaleate ([3H]SCH 23390) were studied in frozen sections of human kidney by using combined in vitro radioreceptor binding and autoradiographic techniques. [3H]SCH 23390 was bound by sections of human kidney in a manner consistent with the labeling of D1 sites, with a Kd value of 3.87 nM and with a Bmax value of 143 fmol/mg protein. Light microscope autoradiography revealed the highest density of [3H]SCH 23390 binding sites within the macula densa and the proximal tubules followed in descending order by the ascending limb of the loop of Henle, the distal tubules and the descending limb of the loop of Henle. No specific binding was noticeable within the glomeruli or within the epithelium of collecting tubules. [3H]SCH 23390 binding sites were also observed within the medial layer of intrarenal artery branches. These findings show that dopamine D1 receptor sites in the human kidney have a localization similar to that described in laboratory mammals with a higher density of tubular than of vascular receptors. The above data account for the vascular, diuretic and natriuretic effects elicited by D1 receptor stimulation in man. PMID- 8423543 TI - Effects of chronic ethanol on growth hormone secretion and hepatic cytochrome P450 isozymes of the rat. AB - Growth hormone (GH) secretion is sexually dimorphic in the laboratory rat and the plasma GH profile is a determining factor in the regulation of the male-specific cytochrome P450 (CYP) 2C11. Acute ethanol has been reported previously to alter the secretion of GH, and in the present investigation, we have studied the effects of chronic (38 days) ethanol on plasma GH profiles, CYP 2C11 and the major ethanol-inducible cytochrome, CYP 2E1, using a total enteral nutrition system, where 35% of the total calories were ethanol. Ethanol-treated rats had elevated (P < or = .05) CYP 2E1 activities and apoprotein levels and increased steady-state mRNA levels encoding for CYP 2E1. Ethanol-treated rats also had reduced (P < or = .05) hydroxylation of testosterone at positions 2 alpha and 16 alpha, lower 2C11 apoprotein levels and lower steady-state mRNA levels encoding for 2C11. In addition, the plasma GH pulse profiles were altered in chronically treated rats by reducing (P < or = .05) the GH pulse amplitude and mean plasma GH concentrations. Our results suggest that: 1) the reduced CYP 2C11 activities, apoprotein levels and steady-state mRNA levels during chronic alcohol exposure are causally related to the alterations in GH secretion; and 2) chronic alcohol exposure elevated CYP 2E1 activities, apoprotein levels and steady-state mRNA levels, and these changes occurred primarily as the result of ethanol rather than undernutrition or as the combination of ethanol and undernutrition. PMID- 8423544 TI - Induction of kidney heme oxygenase-1 (HSP32) mRNA and protein by ischemia/reperfusion: possible role of heme as both promotor of tissue damage and regulator of HSP32. AB - Presently we describe, for the first time, induction of microsomal heme oxygenase 1 (HO-1) mRNA and protein in response to ischemia/reperfusion and therefore define HO-1 as stress protein in the kidney. Specifically, Northern blot analysis of kidneys of rats subjected to bilateral ischemia for 30 min revealed an increase of 8- to 10-fold in the level of 1.8 Kb HO-1 mRNA 6 hr after reperfusion. The increase in transcript level was maintained when assessed after 24 hr. The levels of 1.3 and 1.9 Kb transcripts for the second isozyme of HO, HO 2, were decreased at both time points. The increase in HO-1 mRNA was reflected in HO-1 protein level, as judged by Western blot analysis and at the level of activity as judged by the rate of bilirubin formation. An absence of change in adrenal HO-1 mRNA level subsequent to renal ischemia/reperfusion suggested that the induction of kidney HO-1 did not reflect a generalized response of the rat organs to stress; rather, it was a target organ specific response. Moreover, in kidneys subjected to ischemia 6 and 24 hr after reperfusion, significant increases in the cellular content of heme were observed; heme is a known inducer of HO-1 synthesis. Ischemia/reperfusion also adversely affected concentration of cytochrome P-450 in both mitochondrial and the microsomal fractions of the kidney. We suggest that increase in tissue heme levels may be a significant factor in damage caused by ischemia/reperfusion to renal tissue, whereby the metalloporphyrin promotes oxygen-free radical formation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423545 TI - Complementary deoxyribonucleic acid cloning and expression of a human liver uridine diphosphate-glucuronosyltransferase glucuronidating carboxylic acid containing drugs. AB - A cDNA clone, designated UGT2B7 variant, encoding a 529-amino acid human liver microsomal uridine diphosphate-glucuronosyltransferase (UGT) was isolated from a lambda gt11 human liver cDNA library. UGT2B7 variant synthesized in COS-7 cells was screened for activity toward a range of clinically used drugs and other xenobiotics. The expressed enzyme glucuronidated several carboxylic acid containing nonsteroidal antiinflammatory agents including, in order of relative substrate activity, naproxen, ketoprofen, ibuprofen, fenoprofen, tiaprofenic acid, benoxprofen, zomepirac, diflunisal and indomethacin. Additionally, the stereoselectivity of ketoprofen, naproxen (S/R ratio approximately unity) and ibuprofen (S/R ratio 1.62) glucuronidation by the UGT2B7 variant was shown to differ. Two other carboxylic acid-containing drugs (clofibric acid and valproic acid) and a limited range of drugs containing an alcohol or phenolic functional group were also glucoronidated by expressed UGT2B7 variant. The deduced amino sequence of UGT2B7 variant was shown to differ only in one amino acid (tyrosine for histidine at position 268) from a previously published uridine diphosphate glucuronosyltransferase cDNA, UGT2B7. Like the previously reported enzyme, this variant efficiently glucuronidated hyodeoxycholic acid, estriol, 4-hydroxyestrone and 2-hydroxyestriol. It is, therefore, apparent that UGT2B7 variant has the capacity to glucuronidate with a degree of specificity both endogenous compounds and xenobiotics. Preferred substrates for UGT2B7 variant include xenobiotic carboxylic acids, polyhydroxylated estrogens and hyodeoxycholic acid. PMID- 8423546 TI - Electrophysiological effects of diphenylpyrazolidinone cholecystokinin-B and cholecystokinin-A antagonists on midbrain dopamine neurons. AB - The diphenylpyrazolidinone cholecystokinin (CCK)-B antagonist LY262691 has recently been demonstrated to decrease the number of spontaneously active dopamine (DA) cells in the ventral tegmental area (A10) and substantia nigra (A9) of the anesthetized rat. In the present study, three structural analogs of LY262691 with high selectivity for CCK-B receptors, LY262684, LY191009 and LY242040, also decreased the number of spontaneously active A10 DA cells. Neither an inactive analog (LY206890) nor a CCK-A-selective analog (LY219057) affected the number of spontaneously active A10 DA cells. L-365,260, a benzodiazepine CCK B antagonist, also decreased the number of spontaneously active A10 DA cells. In addition, the more active optical isomer of LY262691 (LY288513) caused twice as large a decrease in the number of spontaneously active A10 DA cells as the less active optical isomer (LY288512). The diphenylpyrazolidinone CCK-B antagonists, but neither the inactive nor the CCK-A selective analog, also decreased the number of spontaneously active A9 DA cells; however, none of these compounds produced catalepsy in awake animals. Single-unit recordings indicated that LY262691 administration inhibited the activity of individual A9 and A10 DA neurons. These results indicate that the firing of A9 and A10 DA neurons is suppressed specifically by antagonism of CCK-B, but not CCK-A receptors. CCK-B antagonists may therefore represent a novel class of antipsychotic drugs. Furthermore, because CCK-B antagonists have no cataleptogenic effects, they may also have a reduced propensity for producing extrapyramidal side effects. In addition, these actions on midbrain DA neurons may contribute to the known anxiolytic activity of CCK-B antagonists. PMID- 8423547 TI - Protective effect of E3330, a novel quinone derivative, in galactosamine-induced hepatitis in rats. AB - The effect of E3330 ((2E)-3-[5-(2,3-dimethoxy-6-methyl-1,4-benzoquinoyl)]-2-nonyl 2-++ +propenoic acid), a novel quinone derivative, was studied in the galactosamine-induced hepatitis model in F344 rats, in which endogenous endotoxin is believed to play a critical pathogenetic role. Subcutaneous injection of 300 mg/kg of galactosamine into rats resulted in liver injury. Oral treatment with E3330 (10-100 mg/kg) 1 hr after galactosamine challenge attenuated the liver injury. E3330 was also effective when administered p.o. 6 or 12 hr after galactosamine challenge. Subcutaneous injection of 1000 mg/kg of galactosamine into rats resulted in more severe liver injury with endotoxemia. The plasma endotoxin was detected 24 to 48 hr after the galactosamine challenge. The time course of increase in plasma endotoxin level was in good agreement with that in plasma aminotransferase activity. E3330 (100 mg/kg) significantly attenuated the liver injury, but did not affect the endotoxin level. Exogenous administration of endotoxin enhanced the hepatotoxicity of galactosamine. Pretreatment with E3330 also protected rats from severe liver injury induced with endotoxin plus galactosamine. These results suggest that E3330 may exert its hepatoprotective effects through inhibition of an effect of endotoxin in galactosamine-induced hepatitis in rats. PMID- 8423548 TI - Effects of heparin, aspirin and a synthetic platelet glycoprotein IIb-IIIa receptor antagonist (Ro 43-5054) on coronary artery reperfusion and reocclusion after thrombolysis with tissue-type plasminogen activator in the dog. AB - Adjuncts to thrombolytic agents have improved coronary patency and prevented early reocclusion after thrombolysis in acute myocardial infarction. The aim of this study was to compare in a canine thrombolysis model the effects of Ro 43 5054 = N-(N-[N-(p-amidinobenzoyl)-beta-alanyl]-L-alpha-aspartyl)-3- phenyl-L alanine-trifluor-acetate, a new glycoprotein IIb-IIIa receptor antagonist with aspirin or heparin. Six groups of 10 dogs each were studied. A platelet-rich coronary thrombus was induced in open-chest dogs by electrical stimulation. In addition to recombinant tissue-type plasminogen activator (30 micrograms/kg.min during 60 min), the dogs received 1) saline, 2) heparin 200 U/kg + 50 U/kg.hr i.v., 3) aspirin 10 mg/kg i.v., 4) heparin+aspirin, 5) Ro 43-5054 (3 micrograms/kg.min) and 6) heparin+Ro 43-5054. The overall reperfusion rate was 70% (range, 60-90%) and comparable in all the six groups. During the 120-min observation period, episodes of reocclusion were observed in the absence of antiplatelet therapy (group 1 and 2) irrespective of heparin treatment. Aspirin prevented coronary reocclusion in half of the reperfused dogs (group 3 and 4). However, after reinforcement of the thrombogenic stimulus, 80% of the reperfused dogs treated with aspirin showed reocclusion, whereas none of them reoccluded when treated with Ro 43-5054. Thus, inhibition of platelet activation by the selective, nonpeptidic glycoprotein IIb-IIIa receptor antagonist Ro 43-5054, although without effect on the time to reperfusion, better protected than aspirin against early reocclusion after thrombolytic therapy. PMID- 8423549 TI - Neurogenic plasma extravasation in the rat nasal mucosa is potentiated by peptidase inhibitors. AB - The increase in vascular permeability associated with neurogenic inflammation in the nasal mucosa is mediated by neuropeptides such as substance P released from sensory nerves. Substance P is degraded by the peptidases neutral endopeptidase 24.11 (NEP-24.11) and angiotensin converting enzyme (ACE). In the present study, we used capsaicin to produce neurogenic inflammation in the nasal mucosa of rats, and we examined the effect of inhibition of NEP-24.11 by phosphoramidon, inhibition of ACE by captopril or inhibition of both enzymes by giving both inhibitors. Using as tracers intravenous Evans blue dye to quantify the extravasation and Monastral blue pigment to localize the sites of leakage, we examined the magnitude and distribution of capsaicin-induced plasma extravasation in the nasoturbinates, maxilloturbinates, ethmoidal turbinates and septum. Capsaicin caused a dose-dependent increase in Evans blue extravasation in the naso- and maxilloturbinates but had only a slight effect in the septum. The leaky blood vessels responsible for this plasma extravasation, as manifested by Monastral blue labeling, were most numerous in the naso- and maxilloturbinates, particularly near the front and free borders. After phosphoramidon, the leakage of Monastral blue was more widespread and extended in a more caudal direction. The response to capsaicin was augmented by phosphoramidon alone but not by captopril alone. However, in the presence of phosphoramidon, captopril further augmented the capsaicin-induced extravasation. We conclude that neurogenic inflammation in the rat nasal mucosa is greatest in the naso- and maxilloturbinates and can be modulated by NEP-24.11 and, to a lesser extent, by ACE. PMID- 8423550 TI - Cholinergic M2 muscarinic receptor-mediated inhibition of endogenous noradrenaline release from the isolated vascularly perfused rat stomach. AB - We measured endogenous noradrenaline (NA) overflow from a vascularly perfused rat stomach in vitro. The stomach was perfused with Krebs-Ringer solution containing 10 microM pargyline. Periarterial nerves, which contain postganglionic sympathetic nerves, around the left gastric artery were stimulated for 1 min with square-wave pulses of 2 msec duration, 2.5 to 5.0 Hz, supramaximal intensity (10 mA). Oxotremorine (10(-8) to 10(-6) M) concentration-dependently inhibited the periarterial nerve stimulation-evoked NA overflow under the presence of 10(-6) M phentolamine. Bilateral vagus nerve stimulation (5 Hz, 2 msec duration, 10 mA, for 1 min) reduced the evoked NA overflow. Oxotremorine (10(-7) M)-induced inhibition of NA overflow was attenuated by atropine, methoctramine (muscarinic M2 receptor antagonist), 4-diphenylacetoxy-N-methylpiperidine (M3 receptor antagonist) and pirenzepine (M1 receptor antagonist) with the following potency; atropine > methoctramine > 4-diphenylacetoxy-N-methylpiperidine >> pirenzepine. The oxotremorine-induced inhibition was attenuated by N-ethylmaleimide (3 x 10( 5) M for 50 min), but was not affected by pertussis toxin pretreatment (10 micrograms/rat, for 4 days). However, this pretreatment with pertussis toxin abolished completely negative chronotropic and inotropic effects of oxotremorine in rat atria. These results suggest that NA release from gastric sympathetic nerve terminals is inhibited by activation of muscarinic M2 receptor, and this receptor-mediated inhibitory mechanisms are insensitive to pertussis toxin. PMID- 8423551 TI - Hormonal and cardiovascular effects of losartan (DuP753), an angiotensin receptor antagonist, in nonhuman primates. AB - Experiments were carried out in conscious dexamethasone-treated cynomolgus monkeys in an attempt to determine if losartan (DuP753) inhibited endogenous as well as exogenous angiotensin II stimulation of angiotensin receptors in vivo. Arterial blood pressure and heart rate were monitored via a radiotelemetry system and blood samples were obtained via vascular access ports for determination of plasma renin activity and plasma aldosterone (PA). Intravenous infusions of angiotensin II elicited reproducible pressor responses and increases in PA. The elevations of blood pressure and PA were completely blocked by losartan (10 mg/kg i.v.). Studies with normal monkeys showed that losartan (10 mg/kg i.v.) elicited a modest hypotension and hyper-reninemia without significantly altering PA. In contrast, studies with sodium-depleted monkeys showed that losartan (1 and 10 mg/kg i.v.) elicited hypotension, a decrease in PA and hyper-reninemia in a dose related manner. Losartan did not significantly alter heart rate in either the normal or the sodium-depleted monkeys. These results were consistent with the conclusion that losartan was an inhibitor of endogenous as well as exogenous angiotensin II stimulation of vascular smooth muscle and adrenal cortical receptors. PMID- 8423552 TI - Optimized thrombolysis of cerebral clots with tissue-type plasminogen activator in a rabbit model of embolic stroke. AB - The dose-dependent effects of tissue-type plasminogen activator (t-PA) on the kinetics of cerebral clot lysis in a rabbit model of middle cerebral artery embolic stroke were investigated. The clots were formed in vitro and tagged with 99Tc for gamma-scintigraphic imaging. After embolization, groups of animals were treated with t-PA. Dose-response curves for the t-PA were generated, and in addition, long and short dosing schedules were assessed. The optimal doses for frequency and rate of cerebral clot lysis in this model are approximately 6.3 mg/kg given over 2 hr or 3.3 mg/kg given over 30 min. These dosing regimens for t PA were accompanied by approximately 50% consumption of plasma plasminogen, fibrinogen and alpha 2-antiplasmin. Doses of t-PA on either side of this optimum caused attenuation in both the frequency and rate of cerebral clot lysis. Treatment with t-PA under either dosing regimen did not augment the frequency of hemorrhagic transformation, but the size of the resultant hemorrhage in those animals where intracranial bleeding occurred was reduced by 3.3 mg/kg t-PA given over 30 min. PMID- 8423553 TI - Pharmacological characterization of the rat hippocampal muscarinic autoreceptor. AB - The interactions of both agonists and antagonists with the hippocampal presynaptic muscarinic autoreceptor were studied in a release assay with mechanically dissociated tissue. Muscarinic agonists inhibited potassium-ion evoked release of [3H]acetylcholine maximally by 30 to 40%. Using partial receptor alkylation, the Kd value for oxotremorine-M was determined to be 2.5 microM, whereas the EC50 for this agonist was 0.43 microM; the Kd/EC50 ratio (5.8) indicates that a modest degree of "spare receptors" exist for this response. The relative intrinsic efficacies of partial agonists (pilocarpine, [(4 hydroxy-2-butynyl)trimethyl ammonium chloride] and bethanechol) were compatible with the involvement of a mixture of the M2 and the M4 receptor subtypes. With Schild analysis, pirenzepine was shown to block with low potency (Ki = 370 nM), which indicated that the M1 muscarinic receptor was not involved in this response. The potencies of the cardioselective drugs (11-[[2 (diethylamino)methyl]-1-piperdinyl] acetyl]-5,11-dihydro-6H-pyrido[2,3 b][1,4]benzodiazepine-6-one] and methoctramine (111 and 61 nM, respectively) were somewhat too low to definitely indicate the involvement of an M2 receptor subtype. The Ki values for the smooth muscle/glandular-selective drugs, 4 diphenylacetoxy-N-methylpiperidine methbromide and hexahydrosiladifenidol (2.5 and 87 nM, respectively), were similar to values for these drugs at the M4 subtype, but their potencies did not clearly distinguish this receptor from the M3 subtype. However, the Kd value for himbacine at this receptor (8.6 nM) ruled out involvement of the M3 receptor.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423554 TI - Species differences in stereoselective metabolism of mephenytoin by cytochrome P450 (CYP2C and CYP3A). AB - Stereoselective involvement of hepatic cytochrome P450 in the metabolism of mephenytoin was investigated in vitro by using livers of five different experimental animal species and humans. The rates of microsomal 4'-hydroxylation were 2 to 6 times higher with the R-enantiomer than the S-enantiomer in rabbits, dogs and rats, whereas the rates of the 4'-hydroxylation in female mice were not different between R- and S-enantiomers. Preferential S-mephenytoin 4' hydroxylation was observed in monkeys as similar to that in humans. The rates of microsomal mephenytoin N-demethylation were approximately 2 times higher with the R-enantiomer than the S-enantiomer in male rats and both sexes of dogs. Antibodies raised against CYP2C11 (anti-CYP2C) clearly inhibited microsomal 4' hydroxylation of S-mephenytoin and N-demethylation of R-mephenytoin in rats, monkeys and humans. Antibodies raised against CYP3A2 (anti-CYP3A) clearly inhibited microsomal 4'-hydroxylation of R-mephenytoin, but marginally S mephenytoin, in rats. Anti-CYP3A, however, showed no clear inhibition on microsomal 4'-hydroxylation and N-demethylation of both enantiomers in monkeys and humans, except for slight inhibition of R-mephenytoin 4'-hydroxylation in male monkeys. The results suggest that stereoselective involvement of rat CYP3A and scant involvement of human CYP3A in R-mephenytoin 4'-hydroxylation are major determinants of the species differences between rats and humans in stereoselective mephenytoin 4'-hydroxylation. PMID- 8423555 TI - 5-HT1A receptors and the tail-flick response. IV. Spinally localized 5-HT1A receptors postsynaptic to serotoninergic neurones mediate spontaneous tail-flicks in the rat. AB - The present study examined the location of the serotonin (5-HT)1A receptors mediating the induction of spontaneous tail-flicks (STFs) in the rat. Serotoninergic neurones were lesioned by i.c.v. administration of 5,7 dihydroxytryptamine, which depleted levels of 5-HT in the spinal cord and other CNS tissues by > 90% without affecting those of noradrenaline and dopamine. In lesioned rats, the ability of the 5-HT releasers, para-chloroamphetamine and methylenedioxymethamphetamine, to elicit STFs was abolished. In contrast, the 5 HT1A agonist, 8-hydroxy-2-(di-n-propylamino)- tetralin (8-OH-DPAT), continued to evoke STFs. In fact, its effect was significantly enhanced in lesioned as compared with sham animals. In (nonlesioned) rats with catheters chronically implanted at the lumbar spinal level, intrathecal 8-OH-DPAT dose-dependently evoked STFs. The action of 8-OH-DPAT was extremely rapid, being maximal within 1 min of injection. Whereas the 5-HT1B/C agonist, TFMPP, the 5-HT1C/2 agonist, DOI, and the 5-HT3 agonist, m-chlorophenylbiguanide, failed to elicit STFs, the action of 8-OH-DPAT was mimicked by several other 5-HT1A agonists: S 14671, 5-MeODMT and lisuride. These also showed a time-course with a rapid onset. Further, the highly hydrophilic 5-HT1A agonist, 5-carboxyamidotryptamine, which fails to pass the blood-brain barrier, likewise dose-dependently elicited STFs upon direct lumbar administration. In contrast, administered onto the cervical spinal cord, it was completely ineffective. Systemic administration of the 5-HT1A antagonists, BMY 7378 or (-)-alprenolol, but not of the 5-HT1C/2 antagonist, ritanserin, nor of the 5-HT3 antagonist, ondansetron, blocked STFs elicited by lumbar administration of 8-OH-DPAT. Conversely, lumbar (but not cervical) administration of BMY 7378 and (-)-alprenolol dose-dependently blocked the action of systemic 8-OH-DPAT. These data demonstrate that STFs in the rat are mediated by 5-HT1A receptors postsynaptic to 5-HT neurones and localized in the lumbar spinal cord. Further, they support the concept of a relationship between STFs and mechanisms of primary sensory (nociceptive) processing. PMID- 8423556 TI - Correlation between number of anastomosed vessels and survival rate in finger replantation. AB - In replantation surgery, an attempt is made to repair as many vessels as possible. However, operative procedures might be simplified to reduce the operative time and the number of personnel required. In an attempt to determine the essential number of vascular anastomoses in finger replantation, the correlation between the number of anastomosed vessels and survival rates was examined in 216 replanted fingers that had been completely amputated. Digits with two arteries repaired had higher survival rates than those with only one artery repaired; however, there was no significant difference between survival rates and the number of arterial anastomoses in any of the zones. The number of anastomosed veins was well-correlated statistically with the survival rate. In Zone I (distal to the lunula), there was no significant difference in the survival rates between no venous anastomosis and one to two venous anastomoses. A statistically significant difference between the survival rates with one venous anastomosis and two or three venous anastomoses was observed only in Zone III (middle phalangeal region). These results support the following conclusions: 1) The essential amount of arterial repair appears to be one arterial anastomosis. 2) The essential venous repair appears to be no veins in Zone I, one vein in Zone II (lunula to distal interphalangeal joint), two veins in Zone III, and one vein in Zone IV (proximal phalangeal region). PMID- 8423557 TI - The results of supraclavicular brachial plexus neurolysis (without first rib resection) in management of post-traumatic "thoracic outlet syndrome". AB - This study evaluated the effectiveness of a supraclavicular brachial plexus neurolysis, without a first rib resection, in relieving the symptom complex traditionally termed "thoracic outlet syndrome." The hypothesis to be tested was that patients with a history of trauma may sustain stretch-type injury and subsequent scarring in and about the brachial plexus which is left untreated during transaxillary first rib resection. This prospective study included 14 patients who each had a neurolysis of the five roots and three trunks of the brachial plexus, plus an anterior scalenectomy through a supraclavicular approach. The results were determined on 11 patients with a mean follow-up of 26.4 months. The results of surgery were five excellent (45 percent), five good (45 percent) and one who failed to improve (10 percent). It is concluded that, with a history of trauma, the symptom complex commonly referred to as "thoracic outlet syndrome" may be primarily due to entrapment of the brachial plexus at sites proximal to the interval between the first rib and the clavicle. It is suggested that: 1) the term "brachial plexus compression" best describes the syndrome without directing the surgeon to remove any one specific anatomic structure and 2) the supraclavicular approach permits excellent surgical exposure of the compressed neurovascular structures. An unexpected observation was the formation of the lower trunk from C8 and T1 proximal to the first rib in the majority of these patients. PMID- 8423558 TI - Experimental study of subcutaneously transferred intestine: effect of the presence of serous membrane on acceptance rate. AB - The serous membrane is considered suspect as one of the obstacles responsible for delaying the acceptance of free jejunal grafts. A model was created of subcutaneous transfer of jejunum from which the serous membrane had been experimentally removed and the authors compared acceptance rates for intestine with and without a serous membrane. Results showed acceptance rates of 0 percent, 37.5 percent, 87.5 percent, and 100 percent respectively, for intestine devoid of serous membrane in which the vascular pedicle had been ligated 4, 5, 7 and 14 days after transfer. When these findings were compared with the results of an experiment on intestine with an intact serous membrane, statistically significant differences were detected between the acceptance rates of the groups ligated 5 and 7 days after transfer. The acceptance time required for intestine devoid of serous membrane was shown to be shorter. PMID- 8423559 TI - Heterotopic finger transfer in ulnar ray deficiency associated with contralateral postaxial polydactyly: a case report. AB - A case of type-I ulnar ray deficiency, combined with postaxial polydactyly of the opposite hand, is described. Heterotopic transfer of the supernumerary finger was performed, in order to improve function in the defective hand and to treat the hexadactyly of the other hand. A combination of these malformations has apparently not previously been reported in the literature. PMID- 8423560 TI - Rat ear transplantation: a feasibility study. AB - The feasibility of rat ear replantation and its inherent advantages as an experimental model have previously been demonstrated. The purpose of this study was to validate the technical feasibility of rat ear transplantation. To eliminate immunologic rejection as a complicating factor, highly inbred Lewis rats (strain LEW/CRIBR) were utilized. The external, internal, or common carotid artery served as the arterial pedicle, and the posterior facial vein as the venous pedicle; tubal cartilage was anastomosed for structural support. Four of five transplanted ears were viable at 10 to 14 weeks postoperatively. The other ear appeared entirely viable, when its host animal died of respiratory complications on postoperative day 2. Rat ears therefore seem to provide a feasible model for transplantation of non-vital organs. PMID- 8423561 TI - The effect of endothelin-1 on muscle microcirculation and its attenuation by verapamil. AB - The vascular endothelium plays an important role in the regulation of vascular tone, by producing both vasodilating and vasoconstricting mediators. Using intravital videomicroscopy, this study examined the in vivo responses of the rat cremaster muscle microcirculation to topical application of endothelin-1 (ET-1), as well as the effect of verapamil pretreatment on these responses. ET-1 produced a potent and persistent vasoconstriction in arteries and veins which lasted for 150 min. The contractile response to ET-1 was significantly more prolonged in arterioles (11 to 30 microns) than in small arteries (40 to 70 microns). Pretreatment of the muscle with verapamil did not inhibit the initial vasoconstrictive action of ET-1, but produced a significant (p < 0.0001) reduction in the recovery time required for vessel relaxation in both small arteries and arterioles. The results suggest that Ca2+ channels contribute to the prolonged vasoconstriction induced in microvessels by ET-1. Because this prolonged vasoconstriction may be responsible for the "no-reflow" phenomenon which occurs in microsurgical procedures, therapy with a calcium antagonist may be useful in attenuating its duration. PMID- 8423562 TI - Successful reconstruction of a high-pressure injection injury of the hand using a first web flap of the foot. AB - High-pressure injection injury to the hand often results in loss of tissue and hand function. The successful reconstruction of a hand following high-pressure injection injury is reported. A free neurovascular flap was transferred from the first web space of the foot to cover a skin defect in the first web space area of the hand. The digital nerves of the thumb and index finger were repaired using sural nerve grafts. PMID- 8423563 TI - Recovery following stretch injury to the sciatic nerve of the rat: an in vivo study. AB - Stretch injuries to peripheral nerves are often associated with poor recovery, which may relate to both the character and the extent of structural alterations in the nerve trunk. Previous biomechanical studies have provided insight into such structural changes; however most of these investigations have involved the testing of specimens after removal from the body. This investigation presents an in vivo model of nerve stretch injury and correlates functional recovery with different regions of the stress-strain relationship described in previous biomechanical studies. Functional recovery was measured for 8 weeks using the Sciatic Functional Index of de Medinaceli. Nerves stretched prior to reaching mechanical failure demonstrated excellent recovery within 2 to 3 weeks, with acute histologic features including variable amounts of degenerating axons. Nerves stretched through the point of mechanical failure were permanently deformed, with widespread degeneration and ruptures in the epineurium and perineurium. Despite these severe pathologic changes, a significant degree of recovery was observed by the end of the study period. These results suggest that if continuity is preserved, a substantial amount of recovery is possible following severe nerve stretch lesions in the rat. Preservation of continuity may create an optimal alignment for regenerating neurons. PMID- 8423564 TI - Adrenal apoplexy--the silent killer. PMID- 8423565 TI - Quality of life measurement in breast cancer. AB - Appropriate assessment of quality of life parameters should be a mandatory requirement when determining the outcome of different treatments for breast cancer. Such measures provide useful, sometimes counterintuitive information concerning treatment costs and benefits and can help guide the clinician with management decisions. It is important to choose well-validated measures of quality of life to enable comparison between studies assessing the impact of different therapeutic modalities and psychosocial interventions. PMID- 8423566 TI - Childhood origins of teenage antisocial behaviour and adult social dysfunction. AB - The main aim of this research was to investigate the childhood predictors (age 8 10 years) of teenage antisocial behaviour (age 18 years) and adult social dysfunction (age 32 years). A sample of 411 London males was followed up from age 8 years to age 32 years. The most important childhood predictors of both outcomes (and of convictions) were measures of economic deprivation, poor parenting, an antisocial family and hyperactivity-impulsivity-attention deficit. However, childhood nervousness and social isolation were negatively related to teenage antisocial behaviour but positively related to adult social dysfunction. It was concluded that the development of adult social dysfunction depended not only on established causes of antisocial behaviour such as economic deprivation and poor parenting but also on causes of internalizing disorders such as childhood nervousness and social isolation. PMID- 8423568 TI - What and why do rheumatologists write? PMID- 8423567 TI - Ultrasonographic appearance of regenerate bone in limb lengthening. AB - The appearance of regenerate bone during the process of limb lengthening using a mono-axial device was studied using radiographs and ultrasound scanning in nine patients. The corticotomy site initially appeared as a sonolucent area. Poorly organized echogenic foci were detectable 2 weeks after the distraction was begun. After 4 weeks, these areas became aligned. At 7-8 weeks, a clear impression of a new cortical margin was seen. The formation of a medullary canal started at this point, progressing to a radiographically evident canal. Ultrasound scanning can reduce considerably exposure to ionising radiation in patients undergoing limb lengthening. Accurate measurements are possible in the early stages, and ossification can be monitored. Axial deviation can be seen but not evaluated. The maturity of the regenerate bone still has to be assessed clinically. PMID- 8423569 TI - Does venous function deteriorate in patients waiting for varicose vein surgery? AB - We have looked at the deterioration in the condition of the lower limbs in a group of 36 patients who were waiting for a median time of 20 months for varicose vein surgery, using clinical examination, colour Duplex scanning and photoplethysmograph (95% refilling times). We found a significant deterioration in this group of patients, with four limbs initially unaffected developing reflux on Duplex scanning, of which three had clinical varicose veins (all four were offered surgery), and of the initial 56 involved limbs, 10 further sources of reflux were found (18%), necessitating alteration of the initial planned surgical procedure. No patient developed deep venous insufficiency or ulceration while on the waiting list, although there was one new case of lipodermatosclerosis. However, had surgery been undertaken after the first assessment, 14 patients (25%) would potentially have required further surgery, although accepting this as justification for allowing patients to wait takes no account of patients suffering or quality of life while waiting for operation. PMID- 8423570 TI - Venous ulceration in males with sex chromosome abnormalities. AB - Two patients with Klinefelter's syndrome and three with 47,XYY who had venous ulcers, and one patient with 47,XYY and a post thrombotic limb are described. Sex chromosome abnormalities should be suspected in tall males with leg ulcers, especially those who have no progeny and are relatively young. Venous ulcers may be more common in males with 47,XYY. PMID- 8423571 TI - The gastroenterology service: a survey of general practitioners' requirements. AB - A postal questionnaire was sent to 634 Leicestershire general practitioners about the service they wanted from their local gastrointestinal unit. Their views were specifically sought in relation to the care of chronic gastrointestinal disorders such as coeliac disease and inflammatory bowel disease. This initial survey was 'testing the water' before addressing GP needs in all areas of gastroenterology including, management issues in peptic ulcer disease and hiatus hernia. The design of the questionnaire was simple with only 12 'yes' or 'no' stems. The response rate to one mailing of the questionnaire was 41% with the rate for each question ranging from 83% (on whether a telephone hot-line would be useful) to 99% (on the value of treatment protocols). There was a poor response rate to some individual stems, with rates of less than 10%, because most GPs only answered 'yes' to the stem they were interested in without answering 'no' to other parts. Most GPs wanted a regular news bulletin on the management of both inflammatory bowel disease and coeliac disease as well as detailed protocols on their treatment. Sixty per cent of respondents wanted a telephone hot line to senior gastroenterologists, with direct dialing to provide immediate advice. Eighty per cent of GPs want shared care with hospital consultants of such patients. A similar proportion thought that this decision should be made jointly by patients and their doctors. There is a clear desire by GPs for a more specialist education in line with the current trend of extending their role. GPs in Leicestershire would value a more active role in the management of patients with chronic intestinal diseases and it is likely that such views are widespread in Great Britain. PMID- 8423572 TI - Do psychiatric patients know what is good for them? AB - The paper reports the findings of a medical audit exercise which monitored 192 consecutive psychiatric admissions. Approximately two-thirds of patients, consultants and referrers provided information on admission objectives and the degree to which they were attained. One-third of the patients furnished information on social problems and satisfaction with treatment. Sanctuary was rated as an important admission objective by patients but neglected by consultants and referrers. Patient satisfaction with treatment was significantly correlated with attainment of patient as well as consultant objectives, and negatively correlated with social problems. Agreement between consultant and patient on admission objectives was inversely related to both duration of contact with psychiatric services and number of previous admissions suggesting that congruence with consultants may be lost over time. The limitations imposed by variable response rates are acknowledged. It is concluded that serious note should be taken of psychiatric patients' own treatment objectives. PMID- 8423573 TI - Do we need computer-based decision support for the diagnosis of acute chest pain: discussion paper. PMID- 8423574 TI - Occupational health--a manager's view: discussion paper. PMID- 8423575 TI - Problems of producing safe poultry: discussion paper. PMID- 8423576 TI - Dr Abraham Gesner: the father of the petroleum industry. PMID- 8423577 TI - A Christmas carol: Charles Dickens and the birth of orthopaedics. PMID- 8423578 TI - Successful but complex drainage of pancreatic abscess. PMID- 8423579 TI - Is it worth writing about? PMID- 8423580 TI - Adult-onset acid maltase deficiency with prominent bulbar involvement and ptosis. PMID- 8423581 TI - Vulval schistosomiasis. PMID- 8423582 TI - Watermelon stomach treated with oral corticosteroid. PMID- 8423583 TI - Reviving the Moore-Federman syndrome. PMID- 8423584 TI - Spontaneous rupture of the femoral arteries. PMID- 8423585 TI - So you want to get it published. PMID- 8423586 TI - Sir Frederick Treves. PMID- 8423587 TI - Cardiopulmonary resuscitation training. PMID- 8423588 TI - Familial precocious puberty in girls. PMID- 8423589 TI - Excessive fear of dilute radiation. PMID- 8423590 TI - Writing papers. PMID- 8423591 TI - Asymmetric synthesis and biological evaluation of beta-L-(2R,5S)- and alpha-L (2R,5R)-1,3-oxathiolane-pyrimidine and -purine nucleosides as potential anti-HIV agents. AB - In order to study the structure-activity relationships of L-oxathiolanyl nucleosides as potential anti-HIV agents, a series of enantiomerically pure L oxathiolanyl pyrimidine and purine nucleosides were synthesized and evaluated for anti-HIV-1 activity in human peripheral blood mononuclear (PBM) cells. The key intermediate 8 was synthesized starting from L-gulose via 1,6-thioanhydro-L gulopyranose. The acetate 8 was condensed with thymine, 5-substituted uracils and cytosines, 6-chloropurine, and 6-chloro-2-fluoropurine to give pyrimidine and purine nucleosides. Upon evaluation of these final nucleosides, the 5 fluorocytosine derivative 51 was found to be the most potent compound among those tested. In the case of 5-substituted cytosine analogues, the antiviral potency was found to be in the following decreasing order: cytosine (beta-isomer) > 5 iodocytosine (beta-isomer) > 5-fluorocytosine (alpha-isomer) > 5-methylcytosine (alpha-isomer) > 5-methylcytosine (beta-isomer) > 5-bromocytosine (beta-isomer) > 5-chlorocytosine (beta-isomer). Among the thymine, uracil, and 5-substituted uracil derivatives, thymine (alpha-isomer) and uracil (beta-isomer) derivatives exhibited moderate anti-HIV activity. In the purine series, the antiviral potency is found to be in the following decreasing order: adenine (beta-isomer) > 6 chloropurine (beta-isomer) > 6-chloropurine (alpha-isomer) > 2-NH2-6-Cl-purine (beta-isomer) > guanine (beta-isomer) > N6-methyladenine (alpha-isomer) > N6 methyladenine (beta-isomer). The cytotoxicity was also determined in human PBM cells as well as Vero cells. None of the synthesized nucleosides was toxic up to 100 microM in PBM cells. PMID- 8423592 TI - Effects of solvation on the ionization and conformation of raclopride and other antidopaminergic 6-methoxysalicylamides: insight into the pharmacophore. AB - Previous work has shown that raclopride in water at neutral pH exists in a zwitterionic form, suggesting a stereoelectronic structure largely different from that of other benzamides. In the present study, the acid-base behavior of other 6 methoxysalicylamides is shown to be comparable to that of raclopride. An extensive investigation by high-temperature molecular dynamics gave insight into the conformational behavior of neutral and zwitterionic raclopride in vacuum and in water. Partitioning of raclopride and a more rigid analogue with characterization (by first-derivative UV spectroscopy) of the predominant forms in the organic phase indicated that only neutral, internally H-bonded forms partition into the organic solvent. Thus, the predominant forms of 6 methoxysalicylamides will be very different in the aqueous and organic phases. In the latter phase, and hence presumably also in the receptor phase, the drugs exist with a neutral, internally H-bonded phenolic group and are therefore stereoelectronically similar to other substituted benzamides. PMID- 8423593 TI - Thromboxane receptor antagonism combined with thromboxane synthase inhibition. 5. Synthesis and evaluation of enantiomers of 8-[[(4-chlorophenyl)sulfonyl]amino]-4 (3-pyridinylalkyl)octanoic acid. AB - The enantiomers of 8-[[(4-chlorophenyl)sulfonyl]amino]-4-(3 pyridinylpropyl)octanoic acid (1) and its pyridinyl ether analog (2) were synthesized using the highly diastereoselective method of alkylation of acyloxazolidinone. These enantiomerically pure compounds were compared with the corresponding racemic compounds 1 and 2 for their in vitro activity. Compounds 1, 1R, and 1S and 2,2S, and 2R were equipotent as thromboxane receptor antagonists (TxRAs) and thromboxane synthase inhibitors (TxSIs) (IC50 = 2-30 nM). Upon oral administration to guinea pigs, the enantiomers inhibited the ex vivo U 46619 induced platelet aggregation with potency similar to that of the corresponding racemic compound. This indicates that the enantiomers have pharmacologic profile and bioavailability similar to that of the corresponding racemic compound. PMID- 8423594 TI - Development of selective tight-binding inhibitors of leukotriene A4 hydrolase. AB - Leukotriene A4 hydrolase is a zinc-containing enzyme which exhibits both epoxide hydrolase and aminopeptidase activities. Since the enzyme product leukotriene B4 is an inflammatory mediator, it is of interest to develop selective inhibitors of leukotriene A4 hydrolase as potential antiinflammatory agents and as mechanistic probes. A systematic study on the enzyme specificity and the inhibition of its amidase activity with more than 30 synthetic inhibitors has led to the development of an alpha-keto-beta-amino ester (26) and a thioamine (27) as tight binding, competitive type transition-state analog inhibitors of the aminopeptidase activity, with Ki values of 46 and 18 nM, respectively. Both compounds also inhibit the epoxide hydrolase activity, with the IC50 values of 1 microM and 0.1 microM for 26 and 27, respectively. PMID- 8423595 TI - (+)-Hemipalmitoylcarnitinium strongly inhibits carnitine palmitoyltransferase-I in intact mitochondria. AB - The reaction of the methyl ester of (R)-norcarnitine with 1-bromo-2-heptadecanone produces (+)-6-[(methoxycarbonyl)methyl]-2-pentadecyl-4,4-dimethylmorpholinium bromide, 3, which hydrolyzes to (+)-6-(carboxylatomethyl)-2-pentadecyl-4,4 dimethylmorpholinium (hemipalmitoylcarnitinium, HPC) upon treatment with aqueous sodium hydroxide. Single-crystal X-ray analyses have confirmed the structures of (+)-HPC and 3. (+)-HPC inhibits carnitine palmitoyltransferase (CPT-I) activity for the forward reaction (palmitoyl-CoA + carnitine-->) in intact mitochondria from rat heart and rat liver. (+)-HPC competitively (versus carnitine) inhibits CPT-I activity in both rat heart and liver mitochondria with Ki = 2.8 +/- 0.5 and 4.2 +/- 0.7 microM, respectively. As one of the strongest specific inhibitors of CPT-I, HPC is a potential therapeutic agent in myocardial ischemia and Type II diabetes. PMID- 8423596 TI - Phenyl-substituted prostaglandins: potent and selective antiglaucoma agents. AB - A series of phenyl-substituted analogues of prostaglandin F2 alpha (PGF2 alpha) were prepared and evaluated for ocular hypotensive effect and side effects in different animal models. In addition, the activity of the analogues on FP receptors was studied in vitro. The results were compared with those of PGF2 alpha and its isopropyl ester. The phenyl-substituted PGF2 alpha analogues exhibited good intraocular pressure reducing effect, were more selective, and exhibited a much higher therapeutic index in the eye than PGF2 alpha or its isopropyl ester. The analogues exhibited high activity on FP receptors in a stereoselective manner for the 15 alpha-hydroxyl group. PMID- 8423597 TI - 7 alpha-Methyl- and 11 beta-ethoxy-substitution of [125I]-16 alpha-iodoestradiol: effect on estrogen receptor-mediated target tissue uptake. AB - The 7 alpha-methyl and 11 beta-ethoxy derivatives of 16 alpha-[125I]iodoestradiol were prepared via halogen exchange with 125I of the corresponding 16 beta bromoestradiol precursors. The 16 alpha-bromo derivatives were obtained via halogenation of the analogous 17-enol acetate, epimerization to the 16 beta isomer, and hydride reduction. Stereochemical assignments were based on high resolution 1H NMR. To evaluate the effect of the nature and stereochemistry of the 16-halo substituent on the relative binding affinity for the estrogen receptor, the analogous 16-chloro derivatives were also prepared. The highest binding affinities were observed with the 7 alpha-methyl-16 alpha-haloestradiols, particularly the bromo and chloro derivatives while the 16 alpha-iodo derivatives gave somewhat lower values. Both the 11 beta-ethoxy and 7 alpha-methyl-16 alpha [125I]iodoestradiols localize in the uteri of immature female rats via a receptor mediated process. Rapid blood clearance of the 125I-labeled 7 alpha-methyl derivative results in lower 125I uptake by the uterus as well as nontarget organs as compared to the 11 beta-substituted estradiol analogs. However, uterus to blood and nontarget ratios are more favorable for the 7 alpha-methyl-16 alpha [125I]iodoestradiol as compared to the analogous 11 beta-ethoxy derivatives suggesting that this compound substituted with 123I may be useful for the in vivo imaging of estrogen receptor-rich breast tumors by single photon emission computerized tomography. PMID- 8423598 TI - Comparative evaluation of seven oligonucleotide analogues as potential antisense agents. AB - 12-Mer analogues, representative of seven different classes of structurally modified oligonucleotides and complementary to the same target, have been compared for their binding affinity for both single-stranded DNA and RNA, resistance to hydrolysis by nucleases in culture medium (RPMI 1640 + 10% inactivated fetal calf serum), and inhibition of HIV-1 replication in de novo infected MT4 T lymphocytes. The viral target was the splice acceptor site of the premessenger coding for the regulatory protein tat. The oligo(2'-O alkyl)ribonucleotides (beta-2'O-allyl-RNA and beta-2'OMe-RNA) were shown to form the most stable hybrids with complementary RNA strands whereas the alpha-anomeric oligodeoxynucleoside phosphorothioate analogue displayed the highest stability in the culture medium. All the modified oligonucleotides examined in the present study exhibited a sequence-nonspecific inhibitory effect on HIV-1 replication, the phosphorothioate analogues being the most active ones (ED50 < 1 microM). PMID- 8423599 TI - Discovery of a novel class of potent HIV-1 protease inhibitors containing the (R) (hydroxyethyl)urea isostere. PMID- 8423600 TI - 3-Tetrahydrofuran and pyran urethanes as high-affinity P2-ligands for HIV-1 protease inhibitors. PMID- 8423601 TI - 2'-(Trimethylammonio)ethyl 4-(hexadecyloxy)-3(S)-methoxybutanephosphonate: a novel potent antineoplastic agent. PMID- 8423602 TI - Active site-directed synthetic thrombin inhibitors: synthesis, in vitro and in vivo activity profile of BMY 44621 and analogs. An examination of the role of the amino group in the D-Phe-Pro-Arg-H series. PMID- 8423603 TI - The morbid anatomy of the human genome: chromosomal location of mutations causing disease. AB - Information is given in tabular form derived from a synopsis of the human gene map which has been updated continuously since 1973 as part of Mendelian Inheritance in Man (Johns Hopkins University Press, 10th ed, 1992) and of OMIM (Online Mendelian Inheritance in Man, available generally since 1987). The part of the synopsis reproduced here consists of chromosome by chromosome gene lists of loci for which there are associated disorders (table 1), a pictorial representation of this information (fig 1a-d), and an index of disorders for which the causative mutations have been mapped (table 2). In table 1, information on genes that have been located to specific chromosomal positions and are also the site of disease producing mutations is arranged by chromosome, starting with chromosome 1 and with the end of the short arm of the chromosome in each case. In table 2 an alphabetized list of these disorders and the chromosomal location of the mutation in each case are provided. Both in the 'Disorder' field of table 1 and in table 2, the numbers 1, 2, or 3 in parentheses after the name of the disorder indicate that its chromosomal location was determined by mapping of the wildtype gene (1), by mapping of the clinical phenotype (2), or by both strategies (3). PMID- 8423604 TI - The clinical features of spondyloepiphyseal dysplasia congenita resulting from the substitution of glycine 997 by serine in the alpha 1(II) chain of type II collagen. AB - The features of a child with spondyloepiphyseal dysplasia congenita resulting from a mutation in one COL2A1 allele were studied. The child was heterozygous for a G to A transition in exon 48 that resulted in the substitution of glycine 997 by serine in the triple helical domain of alpha 1(II) chains of type II collagen. Her longitudinal growth was close to the mean growth curve for children with this chondrodysplasia. Expression of the mutation by chondrocytes would account for the abnormal growth and development of the bones of the limbs and spine. Early expression of the mutation by epithelial cells and later expression by chondrocytes of the developing craniofacial structures would also account for her complex pattern of craniofacial anomalies. The findings in this study confirm that mutations of exon 48 of the COL2A1 gene, that alter the normal Gly-X-Y triplet structure of the corresponding region of alpha 1(II) chains of type II collagen, produce the spondyloepiphyseal dysplasia congenita phenotype. PMID- 8423605 TI - Holoprosencephaly: a family showing dominant inheritance and variable expression. AB - A family with probable dominant holoprosencephaly is presented with five affected subjects in two sibships, the offspring of healthy sisters who are presumed gene carriers. Of the affected children, three had cebocephaly and died shortly after birth. One had left choanal atresia, retinal coloboma, a single central maxillary incisor, microcephaly, short stature, and learning problems. Another had only a single central maxillary incisor. The occurrence of hypotelorism, microcephaly, and unilateral cleft lip and palate as minor manifestations of the gene in possible and probable gene carriers is discussed. PMID- 8423606 TI - Variability of expression in tuberous sclerosis. AB - We present three families in whom a diagnosis of tuberous sclerosis is difficult to secure and we review published reports about similar cases. Tuberous sclerosis has been reported to affect as many as 1 in 9400 subjects in the population. The manifestations of this disease vary not only between but also within families. Currently no reliable method of prenatal diagnosis is available. For these reasons, subjects known to be at 50% risk should be assessed scrupulously to clarify their status. These cases illustrate the difficulties in the clinical diagnosis of tuberous sclerosis and further reinforce the need for a molecular method of determining whether an at risk subject has the disease. PMID- 8423607 TI - Parental age, genetic mutation, and cerebral palsy. AB - Parental age and birth order were studied in 251 patients with cerebral palsy. No parental age or birth order effects were observed in spastic quadriplegia or diplegia, but a paternal age effect was detected in those with athetoid/dystonic cerebral palsy and congenital hemiplegia. These observations indicate that some cases of athetoid/dystonic or hemiplegic cerebral palsy might arise by fresh dominant genetic mutation. PMID- 8423608 TI - Incontinentia pigmenti (Bloch-Sulzberger syndrome). PMID- 8423609 TI - The rapid analysis of dystrophin gene deletions shows variable electrophoretic mobility. AB - The introduction of PCR technology to the molecular diagnosis of genetic diseases has increased the speed and range of DNA tests available. Previous analyses of dystrophin gene mutations were time consuming, taking weeks to complete, and used radioisotopic methods. Further developments in DNA amplification and post amplification techniques have made conventional tube PCR redundant. The rapid methodologies described enable the efficient screening of large populations for genetic disorders, although precautions must be taken when analysing the PCR products. PMID- 8423610 TI - The newly recognised limb/pelvis-hypoplasia/aplasia syndrome: report of a Bedouin patient and review. AB - A Bedouin infant born to consanguineous parents and grandparents is reported. She had Mullerian aplasia and the phenotypic features of the limb/pelvis hypoplasia/aplasia syndrome (MIM 276820). Phenotypic variability of this newly recognised syndrome is briefly discussed. PMID- 8423611 TI - Limb/pelvis-hypoplasia/aplasia syndrome (Al-Awadi/Raas-Rothschild syndrome): report of two Italian sibs and further confirmation of autosomal recessive inheritance. AB - A third family with two Italian neonates affected with limb/pelvis hypoplasia/aplasia syndrome is reported. The disorder shows autosomal recessive inheritance. PMID- 8423612 TI - Mosaicism for duplication 12q (12q13-->q24.2) in a dysmorphic male infant. AB - We report the clinical findings in a boy with mosaicism for a duplication of chromosome 12q13.1-->q24.2. His clinical characteristics are very similar to previously reported mosaic duplications of the distal long arm of 12, as well as several cases with non-mosaic duplications. It is proposed that this represents a clinically distinguishable syndrome for 12q duplication, in mosaic or non-mosaic form. PMID- 8423613 TI - Reproductive possibilities for balanced translocation (14) carriers in families with partial trisomy of proximal 14q. AB - Two cases of 14q proximal partial trisomy in sisters from the same family are reported. Clinical features included craniofacial dysmorphism, skin depigmentation, slight anomalies of the limbs, muscular hypertonia, and physical and mental retardation. The third sister had an abnormal phenotype, different from that of her sibs, and proved to be a carrier of a balanced translocation (2;14)(q36;q21) inherited from their phenotypically normal mother. PMID- 8423614 TI - Delayed speech development, facial asymmetry, strabismus, and transverse ear lobe creases: a new syndrome? AB - A 4 year 9 month old boy and his 3 year 5 month old sister presented with delayed speech development, facial asymmetry, strabismus, and transverse ear lobe creases. The same features were found in their mother, but the father had no such anomalies. To our knowledge this familial association has not been described before and may represent an autosomal dominant syndrome. PMID- 8423615 TI - Arthrogryposis, ophthalmoplegia, and retinopathy: confirmation of a new type of arthrogryposis. AB - Arthrogryposis multiplex congenita is a heterogeneous condition and many different types are clinically recognisable. Recently, a new type of autosomal dominant arthrogryposis was described in a father and son. We report on a male patient with similar clinical features, confirming this distinct type of arthrogryposis. The condition is characterised by congenital contractures of the hands and feet with diminished or absent phalangeal creases, ophthalmoplegia, a rigid trunk, deep set eyes, and (in the oldest patient) an abnormal electroretinogram. Differential diagnosis from amyoplasia, the different types of distal arthrogryposis, and symphalangism is discussed. PMID- 8423616 TI - Waardenburg syndrome and myelomeningocele in a family. AB - We report the first family with Waardenburg syndrome type 1 and myelomeningocele in which more than one subject was affected with both disorders. The possible association is discussed. Prenatal screening for myelomeningocele is suggested for a family with Waardenburg syndrome type 1. PMID- 8423617 TI - Dr. Frank J. Rauscher, Jr.: an appreciation. PMID- 8423618 TI - Prognostic significance of p53 overexpression in node-negative breast carcinoma: preliminary studies support cautious optimism. PMID- 8423619 TI - Three agencies to cooperate in largest agricultural study. PMID- 8423620 TI - EPA finds passive smoking causes lung cancer. PMID- 8423621 TI - Telemedicine: scanning the future of cancer control. PMID- 8423622 TI - Ondansetron leads antiemetic research. PMID- 8423623 TI - Breast cancer care in old age: what we know, don't know, and do. AB - In this review of current pertinent literature from the fields of cancer epidemiology, oncology, health services research, and geriatrics, we describe the epidemiology and unique features of breast cancer and its victims in old age. In addition, we review the current evidence regarding treatment efficacy (i.e., beneficial under ideal circumstances) and effectiveness (i.e., beneficial under usual circumstances) in relation to primary tumor management and the use of adjuvant therapy in early stage disease and outline the challenges associated with studying breast cancer care in older women (> or = 65 years of age). Comorbidity, impaired functional status, lack of social support, and differences in host physiology are among the many factors that influence treatment efficacy and effectiveness, making extrapolation of study findings from younger to older women questionable. Indeed, with the exception of studies of adjuvant tamoxifen therapy, none of the clinical trials supporting the 1990 National Institutes of Health Consensus Development Conference on Treatment of Early-Stage Breast Cancer guidelines have included women over the age of 70 years. Because (a) breast cancer is becoming increasingly common in old age and (b) health-related quality of life is frequently more important to older women than is risk of recurrence or death, all three aspects (surgical management of the primary tumor, postoperative irradiation, and axillary lymph node dissection) of recommended primary treatment deserve fresh scrutiny. The value of adjuvant chemotherapy has yet to be defined. Substantial variations in breast cancer diagnosis, treatment, and care exist, and these differences become greater with increasing age of the patient. However, evidence regarding the reasons for these variations and their relationships with subsequent outcomes is lacking. Challenges for investigators in studies of older women include recruitment into studies, collection of reliable data from interviews or surveys, measurement of disease severity and comorbidity, and selection of relevant outcomes. Given current uncertainty about optimal treatment, clinicians can best serve older patients with early stage breast cancer by involving them in decision-making, taking into account available efficacy data, and individualizing care on the basis of such factors as comorbidity, social support, functional status, and patient preferences for outcomes. Future studies of treatment efficacy in older women should examine the roles of radiation therapy and axillary lymph node dissection that follow breast conserving therapy and should focus on quality of life in addition to recurrence and mortality. Less aggressive treatments, tamoxifen therapy, and adjuvant chemotherapy should also be evaluated. PMID- 8423624 TI - Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer. AB - BACKGROUND: The p53 (also known as TP53) tumor suppressor gene encodes for a nuclear phosphoprotein thought to regulate proliferation of normal cells. Most p53 mutations result in a nonfunctional protein that accumulates in tumor cell nuclei. These common mutations appear to be involved in the development and/or progression of several neoplastic diseases including human breast cancer. PURPOSE: Our purpose was to investigate the relationships between levels of mutant p53 protein expression, tumor cell proliferation rate, and clinical outcome in patients with node-negative breast cancer. METHODS: Expression of mutant p53 protein was evaluated by frozen-section immunohistochemistry (IHC) and light microscopy in 700 breast cancers from axillary lymph node-negative patients with long-term follow-up (median, 54 months). The immunostaining signal was expressed as the sum of scores representing the proportion and staining intensity of negative and positive tumor cell nuclei (ranges, 0 and 2-8, respectively). Statistical comparisons were made between levels of p53 protein expression and disease-free survival, overall survival, and tumor proliferation rate expressed as the percentage of cells in the S phase (%S phase) as determined by flow cytometry. RESULTS: Of the 700 tumors, 362 (52%) showed positive nuclear immunostaining (IHC score > 0). Proliferation rates were significantly higher (P = .0001) in positive tumors (median %S phase, 7.1%) than in negative tumors (4.1%). In a univariate cutpoint analysis, negative tumors (n = 388) versus low positive tumors (IHC score = 2-6; n = 263) versus high-positive tumors (IHC score > 6; n = 99) showed progressively reduced disease-free survival (80% versus 72% versus 58% at 5 years, respectively; P < or = .05 for all pairwise comparisons). Analogous results for overall survival were 88% versus 84% versus 74%; only the result for negative versus high positive tumors was significant (P = .003). In a multivariate analysis, expression of p53 protein and high %S phase were independently associated with reduced disease-free survival (P = .008 and .01, respectively). CONCLUSIONS: Expression of mutant p53 protein was associated with high tumor proliferation rate, early disease recurrence, and early death in node negative breast cancer. Despite the strong direct correlation between accumulation of p53 protein and tumor proliferation rate, both factors were independently associated with poor prognosis, suggesting that p53 may have other biological functions in addition to cell-cycle regulation. IMPLICATIONS: This test, when combined with other prognostic factors, may enhance our ability to identify node-negative breast cancer patients at high risk for early disease recurrence and/or death, for whom the use of adjuvant chemotherapy is unequivocally justified. PMID- 8423625 TI - Human renal carcinoma line transfected with interleukin-2 and/or interferon alpha gene(s): implications for live cancer vaccines. AB - BACKGROUND: Combination therapy with systemically administered interleukin-2 (IL 2) and interferon alpha (IFN-alpha) has resulted in long-term objective remissions in 30% of patients with metastatic renal cell carcinoma (RCC), but toxic effects are clinically significant. PURPOSE: We have thus investigated an alternative therapeutic approach--continuous intratumoral production of IL-2 and/or IFN-alpha by a cytokine-transfected human RCC tumor cell line. METHODS: Plasmid vectors were used to transfect the R11 RCC line with the genes for human IL-2 and/or IFN-alpha by the calcium phosphate precipitation method. Biologic characteristics of the cytokine-transfected tumor cells were determined by assays of thymidine incorporation and cytotoxicity, fluorescence-activated cell-sorter analysis, Northern blotting, and in vivo studies in C3Hf/Sed/Kam mice rendered T cell deficient. RESULTS: The transfected cell lines produced the following amounts of cytokine per 10(6) cells per day: R11-IL-2 (220 U), R11-IFN-alpha (10,240 U), and R11-IL-2 + IFN-alpha (95 U + 1270 U, respectively). Gamma irradiation did not eliminate cytokine secretion. Morphology and growth rates were identical to those for the parental R11 cell line, except for IFN-alpha producing clones, which showed significant growth inhibition. All cytokine producing cells demonstrated increased susceptibility to cell killing by peripheral blood leukocytes (PBL). IFN-alpha producers exhibited enhanced HLA antigen expression and suppressed c-myc messenger RNA expression; when cocultured in vitro, they induced similar changes in parental R11 cells. IL-2 producers could stimulate growth and cytotoxicity of naive (i.e., freshly isolated, uncultured) and activated PBL. All cytokine-producing cells lost their tumorigenicity, as evidenced by failure to grow in the T-cell-depleted mice. When co-injected at a local site but not at a distant site, these cells prevented growth of parental R11 cells. Histologic examination of the injection sites revealed a substantial influx of macrophages. Intraperitoneal administration of IL-2 and/or IFN-alpha could not, however, prevent growth of the parental R11 tumors. CONCLUSION: Local production of high concentrations of IL-2 and IFN-alpha at the tumor site is more effective in preventing tumor growth than systemic administration. IMPLICATION: Continuous local delivery of cytokines via transfer of cytokine genes into tumor cells for use as live cancer vaccines is a novel strategy for manipulation of host-mediated antitumor immune response in patients with advanced RCC. PMID- 8423626 TI - Model-guided determination of maximum tolerated dose in phase I clinical trials: evidence for increased precision. AB - BACKGROUND: A widely used phase I design in clinical trials of chemotherapy for cancer and for AIDS (acquired immunodeficiency syndrome) allows for dose escalation in cohorts of three to six patients. Escalation continues until a predefined percentage of patients experience unacceptable toxic effects at a given dose level. A safe and maximum tolerated dose (MTD) for phase II study is then determined. This standard phase I study design has serious inadequacies. MTD is not a model-based estimate of the true dose that would yield the targeted dose limiting toxicity rate. Moreover, this simplistic study design allows some patients in the phase I study to be treated at doses unlikely to have therapeutic efficacy. PURPOSE: We constructed a novel quantitative assessment design that repetitively evaluates accumulating dose-toxicity data by repeatedly fitting and updating a pharmacodynamic model after small cohorts of patients are treated. The goal was to more accurately estimate the MTD. METHODS: One hundred phase I studies were simulated by both the standard and quantitative assessment phase I designs. We compared determination of MTD, frequency of grade 0 leukopenia (no toxicity), and study size in the studies simulated using the standard design with those in the studies simulated using the quantitative assessment design. RESULTS: The median MTD determined from the 100 studies was nearly identical for the two designs: 100 and 95 mg/m2 per day for standard and quantitative assessment designs, respectively. However, the interstudy variation in the MTD was decreased in the quantitative assessment design. Moreover, the study size was significantly reduced (P < .0001), and the median percentage of patients treated at subtoxic doses (no leukopenia) was significantly lower for the quantitative assessment design (44% versus 48%; P < .0001). CONCLUSION: Our results show clear evidence that a phase I study design using dose and toxicity data in a repetitive and quantitative manner can identify the MTD with more accuracy than the standard design. IMPLICATIONS: New approaches must be explored to improve our ability to identify the optimal dose for phase II studies of chemotherapy for cancer and for AIDS. There is evidence that the quantitative assessment design will identify the MTD with fewer patients, more precision, and fewer patients exposed to suboptimal doses. PMID- 8423627 TI - A prospective cohort study on toenail selenium levels and risk of gastrointestinal cancer. AB - BACKGROUND: Various animal studies and ecologic studies suggest an inverse association between low dietary selenium intake and risk of various types of cancer. PURPOSE: The goal of this prospective cohort study was to investigate the association between toenail selenium levels and risks of stomach cancer and colorectal cancer. METHODS: Our cohort study on diet and cancer started in The Netherlands in 1986 with enrollment of 120,852 subjects aged 55-69 years. Of this number, 58,279 were men and 62,573 were women. Following the case-cohort approach for analysis of the data, we randomly selected from the cohort a subcohort of 3500 subjects (1688 men and 1812 women). After 3.3 years of follow-up, 155 incident cases of microscopically confirmed stomach cancer, 313 cases of colon cancer, and 166 cases of rectal cancer had been detected in the cohort. Toenail selenium data were available for 104 patients with stomach cancer, 234 with colon cancer, and 113 with rectal cancer and for 2459 subjects from the subcohort. RESULTS: In a multivariate analysis, the relative rates (RRs) of stomach cancer for subjects in increasing quintiles of toenail selenium level were 1.00, 0.44, 0.59, 0.84, and 0.64 (trend, P = .491). For men, there was some evidence for an inverse association between toenail selenium levels and stomach cancer: The RR for those in the highest compared with the lowest quintile of toenail selenium was 0.40 (95% confidence interval = 0.17-0.96), but the trend was not statistically significant (P = .136). For stomach cancer in women, there was no negative association with toenail selenium levels. Toenail selenium level was not associated with the risk of colon or rectal cancer. After exclusion of cases diagnosed in the 1st year of follow-up, the RRs of colon cancer for increasing quintiles of toenail selenium were 1.00, 1.27, 1.17, 0.75, and 1.07 (trend, P = .544); for rectal cancer, RR estimates were 1.00, 1.73, 0.83, 1.58, and 1.12 (trend, P = .890). CONCLUSIONS: These data support a suggestive but inconsistent inverse association between selenium levels and risk of stomach cancer. Our findings, like those of other studies, do not suggest an inverse association with risk of colorectal cancer. PMID- 8423628 TI - Mammographic calcifications and risk of subsequent breast cancer. AB - BACKGROUND: Women with proliferative benign breast lesions are at increased risk of breast cancer, and some studies have provided evidence that microscopic calcifications in such lesions enhance the risk. PURPOSE: This study was performed to determine whether calcifications on mammograms are predictive of subsequent breast cancer. METHODS: Data for this study were collected on women enrolled at four of the clinics that participated in the Breast Cancer Detection and Demonstration Project (BCDDP). The presence, morphology, and distribution of calcifications visualized on baseline mammograms for 686 women who developed breast cancer over a 7- to 10-year period of follow-up were compared with those for 1357 controls who remained cancer free. We also compared presence and types of calcifications in breasts in which cancer subsequently developed with those in the contralateral breast. RESULTS: Calcifications were evident at baseline in at least one breast in 381 (55.5%) of 686 cases and in 606 (44.7%) of 1357 controls. The estimated relative risk (RR) of breast cancer was 1.68 in women with calcifications, compared with those having none. There was a statistically significant trend of increasing risk with number of breasts with calcifications; RR increased from 1.28 to 2.14 in women with calcifications in one and both breasts, respectively. In women with unilateral calcifications, RR was greater for the breast in which the calcification occurred (1.48) than for the opposite breast (1.08). The elevated risk persisted for more than 6 years from identification of the calcification, suggesting that these lesions were not indicative of existing carcinomas detected later. Risk was greatest in women with clustered calcifications of any morphology or linearly distributed punctate calcifications (RR = 3.64), and the cancer in women with such calcifications was 4.65 times more likely to occur in the involved breast than in the contralateral breast. Multiple and scattered punctate calcifications, and those of any number or distribution that were ring-shaped or linear, were also associated with subsequent risk of breast cancer (RR = 2.09 and 1.76, respectively) but were not strongly predictive of the side on which the breast cancer occurred. Risk was not altered in women with single punctate or large conglomerate calcifications, although the cancers that subsequently occurred in women with the latter lesions were over three times more likely to develop in the breast with the calcification than in the opposite breast. CONCLUSIONS: These findings are consistent with previously reported relationships between breast cancer and specific histologic types of noninvasive breast lesions. Some types of mammographic calcifications appear to be independent risk factors for breast cancer. IMPLICATIONS: If these results are confirmed by other investigators, mammographic calcifications could serve as an additional indicator of women at high risk for breast cancer who may benefit from intensified follow-up. PMID- 8423629 TI - Enhancement of metastatic potential of murine and human melanoma cells by laminin receptor peptide G: attachment of cancer cells to subendothelial matrix as a pathway for hematogenous metastasis. AB - BACKGROUND: Stable anchorage of circulating cancer cells to the vasculature is a critical step in the formation of hematogenous metastases. Although the basement membrane glycoprotein laminin clearly plays a crucial role in this event, the exact interactive pathways among cancer cells, laminin, and the vessel wall have not been elucidated. In a previous study, we identified synthetic peptide G, which contains the laminin-binding domain of the 67-kd laminin receptor and which inhibits tumor cell adhesion to endothelial cells. PURPOSE: To assess the role of the interaction between laminin and the 67-kd laminin receptor in hematogenous metastasis formation, we studied the effect of peptide G on melanoma cell behavior in vivo and in vitro. METHODS: The effect of peptide G and control peptides was studied in vivo on lung retention and colonizing potential of murine (B16BL6) and human (A2058) melanoma cells injected intravenously in C57BL/6 and nude mice, respectively. In addition, their effect on cell adhesion and chemotaxis to laminin and on binding of iodine 125-labeled laminin to cells was studied in vitro. RESULTS: In vivo, pretreatment of cells with peptide G resulted in a two- to 10-fold significant increase in the number of experimental lung metastases. A significant relative increase in lung retention of peptide G treated tumor cells was observed 48 hours after injection, although after 4 hours a partial reduction was observed. In vitro, peptide G significantly increased laminin binding and cancer cell adhesion to laminin and subendothelial matrix, whereas chemotaxis to laminin was significantly inhibited. CONCLUSIONS: Peptide G differentially affected the biological response of cancer cells to laminin. In vitro, it increased laminin binding and cell adhesion to laminin and subendothelial matrix, whereas it inhibited cell chemotaxis to laminin. In vivo, the overall effect of peptide G was an augmentation of lung metastasis. IMPLICATIONS: Our findings suggest that direct adhesion of tumor cells to the subendothelial matrix is a main pathway for hematogenous metastases and that tumor cell-matrix interaction may be more relevant than tumor cell-endothelial cell attachment in this process. PMID- 8423630 TI - Elevated levels of the angiogenic peptide basic fibroblast growth factor in urine of bladder cancer patients. PMID- 8423631 TI - Radiotherapy for breast cancer. PMID- 8423632 TI - Transient reversion of O4+ GalC- oligodendrocyte progenitor development in response to the phorbol ester TPA. AB - The physiological importance of protein kinase C during oligodendrocyte progenitor maturation was investigated by analyzing the effects of the protein kinase C activator phorbol 12-myristate 13-acetate (TPA) on the morphology, proliferation, and differentiation of oligodendrocytes at sequential stages of development. Monoclonal antibodies A2B5 and O4 were used to identify the A2B5+O4- and the A2B5+O4+ galactocerebroside- progenitor stages. Anti-galactocerebroside and anti-myelin basic protein were used to identify mature, post-mitotic oligodendrocytes. Proliferation was measured by bromodeoxyuridine incorporation. Within 24 hr after addition, TPA induced a down-regulation of the O4 antigen in OL progenitors, and an increase of expression of the intermediate filament protein vimentin, leading to a phenotypic reversion from the vimentin-A2B5+O4+ phenotype to the less mature vimentin+A2B5+O4- stage. Concomitantly, TPA induced an increase in the number of bromodeoxyuridine-labeled oligodendrocyte progenitors and extensive process elongation. The response of O4+ progenitors was transient. Even with continued exposure to TPA, by 4 days after TPA addition the reverted cells ceased proliferation, reacquired O4 immunoreactivity, became vimentin-negative, and began to express galactocerebroside and myelin basic protein, and to display the complex, highly branched morphology characteristic of terminally differentiated oligodendrocytes. These results indicate that modulation of protein kinase C activity by TPA induces a transient reversion of O4+ progenitors to less mature O4- cells, causing a transient inhibition of terminal differentiation. The relationship of these data to similar responses of the OL lineage to specific growth factors and implications for remyelination after pathologic injury are discussed. PMID- 8423633 TI - Expression of the class III beta-tubulin gene during axonal regeneration of rat dorsal root ganglion neurons. AB - The effect of peripheral axotomy on the expression of the class III beta-tubulin gene in adult dorsal root ganglion (DRG) neurons was examined. Of the 5 isotypic classes of beta-tubulin expressed in the mammalian nervous system, only the class III beta-tubulin is neuron specific. While information about the expression of several of the tubulin genes during neuronal development and regeneration has become available recently, very little is known about the expression of beta III tubulin during axonal regeneration. To explore this issue, we examined axotomy induced changes in beta III-tubulin mRNA levels in adult rat lumbar dorsal root ganglion (DRG) neurons at different times (1-28 days) after unilateral sciatic nerve crush using northern blotting of total RNA and quantitative in situ hybridization. These studies showed an initial decrease in beta III-tubulin mRNA levels in axotomized DRG neurons as compared to contralateral controls at 1 day after injury followed by robust increases in beta III-tubulin mRNA levels relative to contralateral controls from 1 to 4 weeks after injury. We postulate that beta III-tubulin may play an essential role in axonal growth because of its unique neuron-specific pattern of expression and its substantial increase in neurons that have been stimulated to regrow their axons. PMID- 8423634 TI - Electrophysiologic and molecular properties of cultured enteric glia. AB - Enteric glia, the support cells of myenteric ganglia, have been widely studied with respect to their morphology and immunohistochemical phenotype, but little is known about their functional properties. We developed a method for the amplification of enteric glia from newborn guinea pigs to further characterize these cells. Treatment with a combination of basic fibroblast growth factor and the adenylate cyclase activator, cholera toxin, permitted expansion of enteric glial cultures to confluence and serial passage for up to 8 months. The long-term cultured cells retained expression of 1) S100 protein, 2) GD3 ganglioside recognized by the monoclonal antibody LB1, and 3) the gene encoding glutamine synthetase. The electrophysiologic properties of cultured enteric glia were studied under whole-cell patch clamp conditions. Most cells expressed "delayed rectifier"-type potassium currents, and some also demonstrated tetrodotoxin sensitive sodium currents. Other subsets of voltage-dependent potassium currents, calcium currents, and glutamate-gated currents were not demonstrable. PMID- 8423635 TI - Transforming growth factor beta as a neuronoglial signal during peripheral nervous system response to injury. AB - In contrast to the central nervous system (CNS), the peripheral nervous system (PNS) displays an important regenerative ability which is dependent, at least in part, on Schwann cell properties. The mechanisms which stimulate Schwann cells to adapt their behavior after a lesion to generate adequate conditions for PNS regeneration remain unknown. In this work, we report that adult rat dorsal root ganglion (DRG) neurons are able, after a lesion performed in vivo or when they are dissociated and cultured in vitro, to synthesize transforming growth factor beta (TGF beta), a pleiotropic growth factor implicated in wound healing processes and in carcinogenesis. This TGF beta is tentatively identified as the beta-1 isoform. Adult rat DRG neurons release a biologically active form of TGF beta which is able to elicit multiple Schwann cell responses including a stimulation to proliferate. Moreover, purified TGF beta-1 produces a Schwann cell morphology alteration and decreases the secretion of tissue-type plasminogen activator (tPA) and enhances the secretion of plasminogen activator inhibitor (PAI) by Schwann cells. This generates conditions which are thought to favor a successful neuritic regrowth. Furthermore, purified TGF beta-1 stimulates type IV collagen mRNA expression in Schwann cells. This subtype of collagen is associated with the process of myelinization. Finally, TGF beta-1 decreases nerve growth factor (NGF) mRNA expression by Schwann cells, an effect which could participate in the maintenance of a distoproximal NGF gradient during nerve regeneration. We propose that neuronal TGF beta plays an essential role as a neuronoglial signal that modulates the response of Schwann cells to injury and participates in the successful regeneration processes observed in the PNS. PMID- 8423636 TI - Early dendrite development in spinal cord cell cultures: a quantitative study. AB - Neurons in dissociated cell culture provide a favorable system for the quantitative analysis of structural changes and the examination of structure function relationships during development. Fragment C of tetanus toxin was used to label neurons in murine spinal cord cell cultures and dendrite outgrowth was monitored by a number of measures. The dissociated neurons increased in morphologic complexity from approximate spheres to highly branched structures during the first week in culture. Much of the structural complexity of the dendrite arbor, as quantified by fractal dimension, was established within 48 hr after plating, i.e., prior to the development of interneuronal contacts. During the first few days in culture, dendrite branching complexity increased more rapidly than dendrite size, whereas after 4 days, fractal dimension remained relatively constant while dendrites continued to grow. Fractal analysis has provided data which suggest that the early development of dendrite branching complexity is determined intrinsically. Fractal dimension, as an effective index of morphologic complexity, should be a useful tool for the further study of extrinsic signals which might modify the generation or stabilization of dendrite form. PMID- 8423637 TI - Congruity of acetylcholine receptor, acetylcholinesterase, and Dolichos biflorus lectin binding glycoprotein in postsynaptic-like sarcolemmal specializations in noninnervated regenerating rat muscles. AB - Noninnervated regenerating muscles are able to form focal postsynaptic-like sarcolemmal specializations either in places of the former motor endplates ("junctional" specializations) or elsewhere along the muscle fibers (extrajunctional specializations). The triple labeling histochemical method was introduced to analyse the congruity of focalization in such specializations of 3 synaptic components: acetylcholinesterase (AChE), acetylcholine receptor (AChR), and a specific synaptic glycoprotein which binds Dolichos biflorus lectin (DBAR). Noninnervated regenerating soleus and extensor digitorum longus (EDL) muscles of the rat were examined and compared with denervated muscles of neonatal and adult rats. All junctional sarcolemmal specializations in noninnervated regenerating muscles accumulated AChE and AChR. Localization of the 2 components was identical within the limits of resolution of the method. DBAR could not be demonstrated in junctional specializations in 17-day-old regenerating muscles. It seems that an agrin-like inducing substance in the former junctional basal lamina invariably triggers the accumulation of both AChE and AChR in the underlying sarcolemma of the regenerating muscle fiber. However, accumulation of DBAR would probably require the presence of the motor nerve. In most of the extrajunctional sarcolemmal specializations in 8-day-old regenerating soleus and EDL muscles, both AChE and AChR accumulated. However, about 10 percent of AChE accumulations lacked AChR and about 35% of AChR accumulations lacked AChE. Even greater variability was observed in 17-day-old regenerating muscles. The presence of DBAR in the extrajunctional postsynaptic-like sarcolemmal specializations could not be demonstrated. Similar extrajunctional sarcolemmal specializations were observed in denervated postnatal rat muscles. About 70% contained both AChE and AChR, and 30% contained only AChR, but none contained DBAR. In denervated mature muscles, sparse extrajunctional AChR accumulations did not contain detectable amounts of AChE. The ability to form complex postsynaptic-like sarcolemmal specializations in the absence of nerve, which is probably inherent to noninnervated immature muscle fibers, may be reduced with muscle maturation. Variable accumulation of individual components in the postsynaptic-like specializations indicates that different triggering factors may be involved in their accumulation or, at least, the mechanisms of their accumulation can function relatively independently. PMID- 8423638 TI - Synaptophysin--a common constituent of presumptive secretory microvesicles in the mammalian pinealocyte: a study of rat and gerbil pineal glands. AB - Recent studies have established that pinealocytes of the mammalian pineal gland contain marker molecules of neuroendocrine cells or paraneurons like the synaptic vesicle-associated protein synaptophysin (p38). The objective of this study was to identify the subcellular synaptophysin-positive compartment and to characterize in detail the intracellular distribution of this protein in rat and gerbil pinealocytes. An analysis of serial semithin sections of plastic-embedded pineals immunostained for synaptophysin, including computer-assisted optical density measurements of synaptophysin immunoreactivities, demonstrated unequivocally that synaptophysin was highly concentrated in dilated process terminals of the pinealocytes. More than 75% of these process terminals were found to border or lie within the pericapillary space. At the ultrastructural level, they contained accumulations of small clear vesicles of variable size that turned out to be the site of synaptophysin immunoreactivity when immunogold staining was performed. In addition, microvesicles surrounding synaptic ribbons were also immunolabeled. Hence, the pinealocyte is the first neuroendocrine cell type that has now been shown to concentrate synaptophysin-positive microvesicles in perivascular process endings. This observation lends strong support to the hypothesis that small clear vesicles in neuroendocrine cells in general, and in pinealocytes in particular, serve secretory functions. The quantitative analysis of completely sectioned process endings revealed that the microvesicles outnumber by far the amount of dense core vesicles and therefore cannot arise by endocytosis of dense core vesicle membranes. Thus, small synaptic-like vesicles probably constitute an independent secretory pathway of the paraneuronal pinealocytes. In the present study, we could also establish the absence of immunoreactivity for synapsin I (belonging to a family of neuron-specific nerve terminal phosphoproteins) from pinealocytes. Synapsin I immunoreactivity was only detectable in intrapineal nerve terminals and varicosities. Taken together, the immunostaining patterns of the pineal gland obtained with antibodies directed against synaptic vesicle-associated proteins render the mammalian pinealocyte a very special type of neuroendocrine cell or paraneuron rather than a "classic" neuron. PMID- 8423639 TI - A piece of my mind. The internist. PMID- 8423640 TI - Health professionals oppose rules mandating participation in executions. PMID- 8423641 TI - Advocates say smoke-free society eventually may result from more curbs, taxes on tobacco use. PMID- 8423642 TI - Multicenter clinical trials of AIDS vaccines scheduled to get under way in coming months. PMID- 8423643 TI - From the Centers for Disease Control and Prevention. Current trends: respiratory syncytial virus outbreak activity--United States, 1992. PMID- 8423644 TI - From the Centers for Disease Control and Prevention. Projections of the number of persons diagnosed with AIDS and the number of immunosuppressed HIV-infected persons--United States, 1992-1994. PMID- 8423645 TI - From the Centers for Disease Control and Prevention. Condom use and sexual identity among men who have sex with men--Dallas, 1991. PMID- 8423646 TI - The supply of rural physicians. PMID- 8423647 TI - The supply of rural physicians. PMID- 8423648 TI - The supply of rural physicians. PMID- 8423649 TI - Does delayed childbearing increase risk? PMID- 8423650 TI - Does delayed childbearing increase risk? PMID- 8423651 TI - Does delayed childbearing increase risk? PMID- 8423652 TI - Cigarette smoking and risk of cataracts. PMID- 8423653 TI - Cigarette smoking and risk of cataracts. PMID- 8423654 TI - Cigarette smoking and risk of cataracts. PMID- 8423655 TI - Cigarette smoking and risk of cataracts. PMID- 8423656 TI - The appropriateness of use of coronary artery bypass graft surgery in New York State. AB - OBJECTIVE: To determine the appropriateness of use of coronary artery bypass graft surgery in New York State. DESIGN: Retrospective randomized medical record review. SETTING: Fifteen randomly selected hospitals in New York State that provide coronary artery bypass graft surgery. PATIENTS: Random sample of 1338 patients undergoing isolated coronary artery bypass graft surgery in New York State in 1990. MAIN OUTCOME MEASURES: Percentage of patients who had bypass surgery for appropriate, inappropriate, or uncertain indications; operative (30 day) mortality; and complications. RESULTS: Nearly 91% of the bypass operations were rated appropriate; 7%, uncertain; and 2.4%, inappropriate. This low inappropriate rate differs substantially from the 14% rate found in a previous study of patients operated on in 1979, 1980, and 1982. The difference in rates was not due to more lenient criteria but to changes in practice, the most important being that the fraction of patients receiving coronary artery bypass grafts for one- and two-vessel disease fell from 51% to 24%. Individual hospital rates of inappropriateness (0% to 5%) did not vary significantly. Rates of appropriateness also did not vary by hospital location, volume, or teaching status. Operative mortality was 2.0%; 17% of patients suffered a complication. Complication rates varied significantly among hospitals (P < .01) and were higher in downstate hospitals. CONCLUSIONS: The rates of inappropriate and uncertain use of coronary artery bypass graft surgery in New York State were very low. Rates of inappropriate use did not vary significantly among hospitals, or according to region, volume of bypass operations performed, or teaching status. PMID- 8423657 TI - The appropriateness of use of percutaneous transluminal coronary angioplasty in New York State. AB - OBJECTIVE: To determine the appropriateness of use of percutaneous transluminal coronary angioplasty (PTCA) in New York State. DESIGN: Retrospective randomized medical record. SETTING: Fifteen randomly selected hospitals in New York State that provide PTCA. PATIENTS: Random sample of 1306 patients undergoing PTCA in New York State in 1990. MAIN OUTCOME MEASURES: Percentage of patients who underwent PTCA for indications rated appropriate, uncertain, and inappropriate. RESULTS: The majority of patients received PTCA for chronic stable angina, unstable angina, and in the post-myocardial infarction period (up to 3 weeks). Fifty-eight percent of PTCAs were rated appropriate; 38%, uncertain; and 4%, inappropriate. The inappropriate rate varied by hospital from 1% to 9% (P = .12); the uncertain rate, from 26% to 50% (P = .02); and the combined inappropriate and uncertain rate, from 29% to 57% (P < .001). There was no difference in appropriateness when the institutions were grouped by volume (fewer than 300 procedures annually or at least 300 procedures annually), location (upstate vs downstate), or by teaching status. CONCLUSIONS: Few PTCAs were performed for inappropriate indications in New York State. However, the large number of procedures performed for indications that were rated uncertain as to their net benefit requires further study and justification at both clinical and policy levels. PMID- 8423658 TI - The appropriateness of use of coronary angiography in New York State. AB - OBJECTIVE: To determine the appropriateness of use of coronary angiography in New York State. DESIGN: Retrospective randomized medical record review. SETTING: Fifteen randomly selected hospitals in New York State that provide coronary angiography. PATIENTS: Random sample of 1335 patients undergoing coronary angiography in New York State in 1990. MAIN OUTCOME MEASURES: Percentage of patients who underwent coronary angiography for appropriate, uncertain, or inappropriate indications. RESULTS: Approximately 76% of coronary angiographies were rated appropriate; 20%, uncertain; and 4%, inappropriate. Inappropriate use did not vary significantly between the elderly (ie, patients aged 65 years and older) and nonelderly, 4.7% and 3.9%, respectively. Although the rate of inappropriate use varied from 0% to 9% among hospitals, the difference was not significant. Rates of appropriateness did not vary by hospital location (upstate vs downstate), volume (fewer than 750 procedures annually or at least 750 procedures annually), teaching status, or whether revascularization was available at the hospital where angiography was performed. CONCLUSIONS: Although coronary angiography was used for few inappropriate indications in New York State, many procedures were performed for uncertain indications in which the benefit and risk were approximately equal or unknown. PMID- 8423659 TI - The epidemiology of bacillary angiomatosis and bacillary peliosis. AB - OBJECTIVE: To determine environmental risk factors for bacillary angiomatosis bacillary peliosis (BAP), and to confirm infection with Rochalimaea species. DESIGN: Case-control study. SETTING: Community and university hospitals and clinics. PATIENTS: Case patients (N = 48) had biopsy-confirmed BAP. Controls (N = 94) were matched to patients by institution and by human immunodeficiency virus (HIV) serological status. MAIN OUTCOME MEASURES: Clinical information was obtained from medical records. Subjects were queried about environmental exposures. Univariate odds ratios (ORs) with 95% confidence intervals (CIs) were determined. Bivariate analyses were performed on variables associated with disease by univariate analysis. DNA from 22 available case-patient tissues and from 22 control tissues was amplified with the polymerase chain reaction (PCR) using primers designed to detect Rochalimaea species. RESULTS: We identified five HIV-negative, immunocompetent case patients; one HIV-negative, immunodeficient case patient; and 42 HIV-positive case patients. There were no significant differences between case patients and controls by race, sex, age, or risk factors for HIV infection. Owning a cat (OR, 2.8; CI, 1.4 to 5.8) and history of a recent cat lick (OR, 1.95; CI, 1.0 to 3.8), cat scratch (OR, 3.7; CI, 1.7 to 8.0), or cat bite (OR, 3.9; CI, 1.8 to 8.9) were associated with disease in the univariate analysis. In bivariate analyses, only the variables representing traumatic contact with a cat (bite or scratch) remained associated with disease. No other environmental exposure was associated with disease. The PCR amplified a DNA fragment of the size expected for Rochalimaea species in all 22 case-patient tissue specimens. CONCLUSIONS: These data suggest that BAP is a new zoonosis associated with both traumatic exposure to cats and infection with Rochalimaea species or a closely related organism. PMID- 8423660 TI - Privacy beliefs and the violent family. Extending the ethical argument for physician intervention. AB - Privacy beliefs associated with the family impede physicians' response to domestic violence. As a private sphere, the family is regarded as sacred, separate, and hidden from public view. Hence, physicians who look for or uncover violence in the family risk defilling a sacred object and violating norms of non interference. Privacy beliefs also obfuscate the ethical analysis of physicians' duties to intercede on behalf of battered patients. Ethical principles of beneficence and nonmaleficence have been invoked to justify physicians' duties to abused patients; however, the principle of justice has not been invoked. Ethical analysis of physicians' duties in this area must be broadened to include the principle of justice. Justice is at stake because establishing conditions favorable to self-respect is a requirement of justice, and the response physicians make to battered patients carries important ramifications for supporting patients' self-respect and dignity. If justice forms part of the ethical foundation for physician intervention in domestic violence, mandatory steps that do not transgress the confidentiality of the physician-patient relationship or infringe the patient's autonomy should be taken, such as requiring domestic violence training in medical education and following treatment plans and protocols to identify abuse and provide assistance to battered patients. PMID- 8423661 TI - Who pays for published research? AB - OBJECTIVE: To determine the extent of unfunded research published in major medical journals. DESIGN: Review of original research completed in the United States and published in 23 official journals of internal medicine and neurology during 1 month in 1991. Investigators were contacted to confirm lack of funding. MAIN OUTCOME MEASURE: Percentage of unfunded, published original research. RESULTS: One hundred ninety-six articles were evaluated. There was at least one unfunded study in 78% of journals. Forty-five published studies (23%) were unfunded. Among those 45 studies, 7% were clinical trials, 9% were cohort studies, 18% were cross-sectional or case-control studies, 53% were case series, and 13% were surveys. Thirteen unfunded studies involved procedures that presumably were performed for research purposes and not as part of routine patient care. CONCLUSIONS: Nearly one quarter of original research published in major medical journals was unfunded. Seven percent of published research involved direct clinical costs that were not accounted for by the investigators. These costs may have been passed on to study participants or third-party payers. PMID- 8423662 TI - Intensive care, survival, and expense of treating critically ill cancer patients. AB - OBJECTIVE: To determine the survival and factors affecting the survival of patients with solid tumors and hematologic cancers who were admitted to the intensive care unit, the time these patients spent at home (meaningful survival) before they died, and the cost per year of life gained and per year of life gained at home. DESIGN: Survival and cost-effectiveness analysis. SETTING: A tertiary-care cancer center at a university medical center. PATIENTS: Every patient admitted to the intensive care unit between July 1, 1988, and June 30, 1990, was entered into the study. This group comprised 83 patients with solid tumors and 64 patients with hematologic cancers. MAIN OUTCOME MEASURES: Factors affecting survival, such as age, sex, malignancy, length of stay in the intensive care unit, and necessity for mechanical ventilator assistance, as well as cost per year of life gained and cost per year of life gained at home. RESULTS: The only factor that significantly affected survival was the requirement for mechanically assisted ventilation for patients with hematologic cancers. More than three fourths of the patients in either group spent less than 3 months at home before dying. The cost per year of life gained for patients with solid tumors was $82,845 and for patients with hematologic cancers was $189,339. The cost per year of life gained at home was $95,142 for patients with solid tumors and $449,544 for patients with hematologic cancers. CONCLUSION: The majority of patients with solid tumors and hematologic cancers admitted to the intensive care unit die before discharge, or, if they survive the hospital admission, they spend a minimal amount of time at home before dying. This limited survival is achieved at considerable cost. Physicians who treat patients with neoplastic disease should discuss potential outcomes and the possibility of withdrawing life supportive therapy if appropriate with the patient and family, so that a reasonable strategy can be agreed on before the initiation of therapy. PMID- 8423663 TI - Physician utilization disparities between the uninsured and insured. Comparisons of the chronically ill, acutely ill, and well nonelderly populations. AB - BACKGROUND: This study examines the associations between lack of health insurance coverage and physician utilization for the chronically ill, acutely ill, and well nonelderly populations in the United States. METHODS: Cross-sectional data from the 1989 National Health Interview Survey, conducted by the National Center for Health Statistics, were analyzed for the nonelderly population using a correlational, two-group design (N = 102,055). Analytic models, using multiple logistic regression, were tested to predict the odds and likelihood of physician utilization for the uninsured and insured in the three subpopulations (ie, chronically ill, acutely ill, and well), controlling for health status, number of conditions, and geographic, sociodemographic, and economic factors. Disparities in utilization were then calculated between the uninsured and insured for each subpopulation. RESULTS: The nonelderly uninsured were consistently less likely than the insured to have received any health care within 12 months. Moreover, there were differential effects of being uninsured on utilization depending on whether an individual was chronically ill, acutely ill, or well. Whereas chronically ill and well uninsured persons were half as likely to have seen a physician as their insured counterparts (odds ratio, 0.50), acutely ill uninsured persons were almost two thirds as likely to receive physician care (odds ratio, 0.62). Thus, the disparity in physician utilization between the uninsured and insured was larger for the chronically ill and well than for the acutely ill; uninsured acutely ill were less likely to go without care. Of the three populations, those in the well population had average disparities with the largest magnitude (40%), compared with disparities of the chronically ill (20%) and acutely ill (10%). CONCLUSIONS: These disparities represent large inequities in utilization of care by the uninsured, particularly for the chronically ill and well. Whether these disparities result from lower access or individual choice cannot be determined from this study. When viewed in light of other studies examining the impact of utilization on health status, these results provide support for the development of comprehensive health insurance packages with universal coverage and better inclusion of chronic and preventive care models in benefit packages. PMID- 8423664 TI - Removing the incentive to sell kids tobacco. A proposal. PMID- 8423665 TI - Toward fewer procedures and better outcomes. PMID- 8423666 TI - Eustachian tube dysfunction. PMID- 8423667 TI - Vaccinations for travelers. PMID- 8423668 TI - [Transaction of the 80th Annual Meeting of the Japanese Urological Association]. PMID- 8423669 TI - Genes in a bottle. PMID- 8423670 TI - Cellular stress response induces selective intranuclear trafficking and accumulation of morbillivirus major core protein. AB - BACKGROUND: Using canine distemper virus (CDV) encephalomyelitis as a model for morbillivirus-induced intranuclear inclusion body (INB) formation, we previously demonstrated that the cellular basis for INB within infected astrocytes are viral antigen-associated nucleolar derivatives known as complex nuclear bodies. Here, we examined the relationship between the cellular stress response and intranuclear CDV protein trafficking so that we may begin to define cellular events leading to formation of complex nuclear bodies containing viral protein. EXPERIMENTAL DESIGN: Confocal dual-label immunofluorescence microscopy was used to simultaneously determine the distribution of CDV antigen and the major inducible 70 kilodalton (kd) heat shock protein (i.e., 72 kd heat shock protein (HSP)) within infected canine central nervous tissue. This dual-label immunofluorescent technique was also used to test the relationship between cellular stress and CDV core protein distribution in a CDV persistently infected cell line. RESULTS: The cytoplasm of INB-positive astrocytes expressed elevated 72 kd HSP characteristic of the cellular stress response; INB contained 72 kd HSP and the major CDV nucleocapsid protein (i.e., N). Intranuclear inclusion bodies did not stain positively for the CDV nucleocapsid phosphoprotein nor was intranuclear N protein observed in the absence of nuclear or elevated cytoplasmic 72 kd HSP. In the persistently infected cell line, induction of the cellular stress response elevated 72 kd HSP and produced translocation of the CDV N protein to the nucleus from its normal location in the cytoplasm of unstressed infected cells. As observed in vivo, the transport was specific to N and was correlated to the induction of complex nuclear bodies. The latter was documented by transmission electron microscopy, where complex nuclear bodies formed in shocked infected cells, but not shocked uninfected cells or nonshocked infected cells. CONCLUSIONS: CDV infection in vivo induces the cellular stress response. Paradoxically, this normally protective cellular response mediates redistribution of viral N protein from the cytoplasm into infected cell nuclei, which is the likely basis for complex nuclear body formation and a unique form of virus induced cytopathic effect. PMID- 8423671 TI - Radiation-induced increased platelet-activating factor activity in mixed saliva. AB - BACKGROUND: Platelet-activating factor (PAF), a family of structurally-related phospholipid mediators of inflammation, is present in normal human mixed saliva; however, its role in oral biology and the homeostasis of oral host defense mechanisms remains to be established. EXPERIMENTAL DESIGN: The current study was designed to evaluate the salivary levels of PAF in patients with oral mucositis that developed as a complication of head and neck irradiation for oral cancer. PAF activity was assessed in platelet bioassay and expressed relative to the activity of authentic PAF, 1-O-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine (C16:0-AGEPC). RESULTS: A significant increase in salivary PAF levels was observed in patients with mucositis (47,032 +/- 12,731 C16:0-AGEPC fmole equivalents/ml of saliva, mean +/- SE, N = 7) as compared with normal subjects (5,568 +/- 1,135 C16:0-AGEPC fmole equivalents/ml of saliva, N = 27). Phospholipid fractionation of the PAF isolated from the saliva of patients with mucositis by reverse phase high performance liquid chromatography revealed a single peak of activity that corresponded with the elution profile of C16:0 AGEPC, the most biologically active molecular species of PAF. In contrast, the PAF isolated from normal human mixed saliva contained multiple molecular species of PAF. CONCLUSIONS: These results suggest that this potent phospholipid inflammatory mediator may play a role in the inflammation and tissue injury associated with mucositis resulting from radiation treatment for oral cancer. PMID- 8423672 TI - Direct myocardial transfection in two animal models. Evaluation of parameters affecting gene expression and percutaneous gene delivery. AB - BACKGROUND: Gene therapy represents a novel approach to the treatment of a variety of disease states. Direct injection of pure untreated DNA into skeletal and cardiac muscle is sufficient to perform gene transfer in vivo. Little information is available, however, regarding the extent to which individual parameters of direct gene delivery affect the efficiency of myocardial transfection. Moreover, the fact that all previously reported studies of successful myocardial transfection were performed using open chest surgery to visualize the heart for direct injection of DNA constitutes a potential limitation to clinical applications of myocardial gene transfer. The objectives of the present study were: (a) to determine the extent to which gene expression is altered by varying the amount of DNA and the volume of the delivery vehicle; (b) to study whether protracted expression of DNA persists after direct myocardial transfection; and (c) to test the feasibility of percutaneous, transthoracic myocardial injection of DNA. EXPERIMENTAL DESIGN: New Zealand white rabbits (N = 30) were transfected with the firefly (Photinus pyralis) luciferase reporter gene. Twenty-three rabbits were segregated into 5 groups according to the amount of DNA and volume of delivery vehicle injected. These rabbits were sacrificed at 5 days posttransfection to measure the level of luciferase gene expression. Three more rabbits were designated exclusively for in situ hybridization. Two rabbits were sacrificed at 5 and 6 months, respectively to evaluate long-term expression. Two additional rabbits were used as controls. Yucatan microswine (N = 4) were used to evaluate the feasibility of percutaneous, transthoracic myocardial transfection with beta-galactosidase or luciferase reporter genes. Microswine were sacrificed immediately post-sham transfection (N = 2), or 5 days postmyocardial transfection (N = 2). RESULTS: Among 11 rabbits, transfection with 10 micrograms of DNA (N = 4), 25 micrograms of DNA (N = 3), and 50 micrograms of DNA (N = 4) in 100 microL of injectate yielded a step-wise but statistically insignificant increase in luciferase activity. Among 16 rabbits, transfection with 50 micrograms of DNA in injectate volumes of 50 microliters (N = 4), 100 microliters (N = 4), 150 microliters (N = 4), and 300 microliters (N = 4) yielded a statistically significant (p < 0.05) increase in luciferase activity in those hearts transfected with 300 microliters compared with those hearts transfected with either 50 or 100 microliters. Luciferase activity at 5 and 6 months postmyocardial transfection was 7.14 and 68.8 Turner light units, respectively. In situ hybridization confirmed that myocytes represented the site of luciferase expression. Percutaneous myocardial transfection was successfully accomplished with both reporter genes. CONCLUSIONS: The results of the present study demonstrate that increasing the amount of DNA employed for direct myocardial transfection fails to produce a statistically significant increase in the level of gene expression. In contrast, increasing the volume of injectate used to directly transfect a constant amount of DNA (50 micrograms) produced significantly augmented expression of the reporter luciferase gene. Luciferase expression was detected 6 months posttransfection. Successful transfection after fluoroscopic-guided direct percutaneous delivery suggests that it may be feasible to deliver genetic material into the myocardium of patients in a similar fashion. PMID- 8423673 TI - Squamous metaplasia expression of proto-oncogenes and P 53 in lung cancer patients. AB - BACKGROUND: The question of whether bronchial squamous metaplasia is a true preneoplasia is important and demonstrated in animal for several carcinogens. We have now approached this problem in humans and in vivo. EXPERIMENTAL DESIGN: Squamous metaplasia in the close vicinity of surgically resected lung tumors were evaluated for their mitotic index and screened for proto-oncogenes and P 53 protein expression by immunohistochemistry and/or in situ hybridization. RESULTS: Among 16 patients, 4 had squamous metaplasia positive for either myc messages and/or for P 53 protein accumulation. In the same patients (3/4), the autologous bronchial tumors were also positive for the same markers. Squamous metaplasia positivity was observed essentially in patients with advanced diseases and only in squamous cell carcinomas. In addition, when evaluated with 5 iodo-2' deoxyuridine systemic infusion, all patients presented hyperproliferative basal squamous metaplasic cells. CONCLUSIONS: These results are reminiscent of the typical preneoplastic changes observed in familial colic adenomatosis, where genetic changes accumulate in hyperproliferative cells. They also suggest that bronchial squamous metaplasia could be an authentic preneoplasia in, at least, squamous cell carcinomas. PMID- 8423674 TI - Early alterations in ras protooncogene mRNA expression in testosterone and estradiol-17 beta induced prostatic dysplasia of noble rats. AB - BACKGROUND: The simultaneous treatment of intact Noble rats with testosterone and estradiol-17 beta for 16 weeks consistently induces intraductal dysplasia exclusively in the dorsolateral lobe (DLP) of the prostate. The lesion closely resembles human prostatic dysplasia and is considered to be a preneoplastic alteration, since invasive carcinoma frequently develop after long-term treatment of rats with both steroids. In our current study, we investigated steady-state ras transcript expression at the earliest recognized stages of sex steroid induced dysplasia in the DLP. Our interest in studying ras expression in these evolving lesions stems from the pivotal role this family of genes are thought to play in the regulation of cell division and differentiation as well as in the genesis of a variety of human and animal neoplasms. EXPERIMENTAL DESIGN: Northern blotting and in situ hybridization were used to study ras protooncogene mRNA expression in the DLPs of NBL rats harboring sex steroid-induced ductal dysplasia and to compare findings with those from prostates of castrated and castrated androgen-treated animals. Since the prostate is an androgen-dependent gland, alterations in ras expression were compared with changes in the transcript levels of two androgen-responsive genes that encode for a prostatic secretory protein, seminal vesicle secretion protein II, and the androgen receptor. RESULTS: Similar to the situation for androgen receptor expression, orchiectomy initially enhanced levels of both H- and K-ras transcripts, whereas T administration to castrates was found to return the values to levels found in intact rats. Sixteen weeks of T and E2 administration to intact rats caused levels of H-ras mRNA and a 2.4 kb K ras transcript to rise by 50 and 60%, respectively in the DLPs with dysplasia when compared with counterpart lobes from untreated control animals. In situ hybridization revealed markedly enhanced H-ras expression in some dysplastic DLP foci and no changes in histologically normal ducts and acini. CONCLUSIONS: Taken together, results from our studies suggest that the enhanced focal expression of ras protooncogenes may participate in early aberrant proliferation of prostatic ductal cells of the DLP. Early alterations of ras expression in dysplastic lesions may therefore be a key contributing event in the multistage development of prostate cancer in this animal model. PMID- 8423675 TI - Role of integrins and other cell adhesion molecules in tumor progression and metastasis. AB - Despite rapid advances in our understanding of the biology of cell adhesion, the data available in the literature make it is difficult to propose one simple scheme in which cell adhesion molecules can be related to tumor growth and metastasis. This difficulty can be related to a number of factors. Some of the apparently conflicting experimental results that demonstrate both enhanced or diminished tumor cell adhesion during tumor progression may be attributed to the experimental systems used. Those studies that have injected tumor cells intravenously have, in general, shown that enhanced tumor cell adhesiveness correlates with metastatic ability. It should be recognized that this experimental approach bypasses many of the early stages of the metastatic cascade and is biased towards tumor cells with an enhanced ability to form aggregates with cells in the circulation and to adhere to distant vascular sites. On the other hand, studies that have implanted tumors into animals and allowed them to grow and metastasize (spontaneous metastasis) have generally demonstrated an inverse relationship between adhesive ability and the ability to metastasize. Another major obstacle in understanding the role of CAMs in metastasis is the well known problem of tumor heterogeneity and the phenotypic instability of metastatic cells over relatively short periods of time (141). The cells that make up a metastatic focus may thus be quite different from the tumor cells that originally formed the lesion. It is quite possible that the selective pressures that initially enable a cancer cell to form a metastatic lesion may be quite different than those that later favor rapid tissue growth. The major obstacle in making any sweeping generalizations about cell adhesion molecules and tumor progression, however, is that the process of successful metastasis is inherently complex, requiring tumor cells to possess decreased adhesive interactions with surrounding cells and extracellular matrix at some points in the cascade and increased adhesive interactions at other times. Based on the information available, the following scenario can be proposed. Using the schema shown in Figure 1, successful metastasis initially requires that normal cell-cell and cell substratum adhesion be disrupted, causing release of neoplastic cells from the primary tumor (step 1). For epithelial tumors, down-regulation of cadherins and perhaps, integrins, appear to be involved. This loss of cell adhesion must be followed by migration of tumor cells into the vascular system (step 2), a step requiring efficient cell-substratum interactions. In melanomas, this step seems to require expression of the vitronectin receptor.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423676 TI - Ultrastructural changes of extracellular matrices in diabetic nephropathy revealed by high resolution scanning and immunoelectron microscopy. AB - BACKGROUND: Diabetic nephropathy is invariably associated with proteinuria. EXPERIMENTAL DESIGN: To delineate the mechanism(s) of proteinuria in diabetic nephropathy, ultrastructural changes of the glomerular basement membranes (GBMs) were studied by high resolution scanning and immunoelectron microscopy. Acellular glomeruli from diabetic and age-matched control human subjects were prepared by detergent method and subjected to conductive staining, the technique in which the tissues are impregnated with metals rather than surface-coated with metallic alloys for visualization by electron microscopy. Subsequent to conductive staining, the tissues were examined by in-lens field emission scanning electron microscopy. RESULTS: Thirty glomeruli, each from the control and diabetic groups, were examined by scanning microscopy. In diabetic GBMs, a loose meshwork structure consisting of numerous pores of approximately 8 nm in diameter and distinct strands was observed. In contrast, meshwork structure was not readily discernible in controls and a few pores were observed. Five glomerular capillary loops, each from control and diabetic groups, were examined by immunoelectron microscopy. In controls, heparan sulfate-proteoglycan was localized in the lamina rara interna and externa, and type-IV collagen was distributed throughout the whole width of the GBM. In diabetic GBMs, a relative loss of staining of heparan sulfate-proteoglycan, both in the lamina rara interna and externa of the GBM, was observed. Type IV collagen was distributed in all layers of the thickened GBM, and the absolute number of the immunogold particles was increased. However, immunogold particle density of type IV collagen per unit area was decreased as compared with the control. CONCLUSIONS: These ultrastructural and immunoelectron microscopic changes in the GBM may explain the loss of charge as well as size selectivities of the glomerulus, as observed in diabetic nephropathy associated with proteinuria. PMID- 8423677 TI - Stereological estimates of volume-weighted mean glomerular volume in streptozotocin-diabetic rats. AB - BACKGROUND: The stereological estimate of mean glomerular volume is an objective and reproducible method of measuring glomerular size in terms of absolute volume. This method provides objective, unbiased, and reproducible data that may be used in the description of glomeruli. EXPERIMENTAL DESIGN: We have made a morphometric analysis of glomerular size in three cortical areas (superficial, midcortical, and juxtamedullary) in normal and diabetic animals, using the point-sampled intercepts method. The kidney weight and volume as well as the cortical and glomerular volume density were also calculated. Our study included 35 animals (5 control and 30 experimental). Fifteen experimental animals were treated with 50 mg/kg streptozotocin dissolved in citrate buffer at pH 4.5. The other 15 animals were sham injected with citrate buffer. In all cases the tissues were fixed in 10% formaldehyde and embedded in paraffin. Sections were cut at 4 microns and stained with periodic acid-Schiff and a systematic sampling was performed. RESULTS: In the control and experimental groups the juxtamedullary glomeruli was significantly larger than superficial and midcortical glomeruli. Mean values of the kidney weight and volume and the volume-weighted mean glomerular volume show a significant increase at 4, 20, and 50 days after streptozotocin-induced diabetes. CONCLUSIONS: The increase in the volume-weighted mean glomerular volume was similar in the three cortical areas, thus suggesting that the effect of diabetes on glomerular enlargement is similar in all cortical areas. PMID- 8423678 TI - Glomerulosclerosis and body growth are mediated by different portions of bovine growth hormone. Studies in transgenic mice. AB - BACKGROUND: Mice transgenic for bovine growth hormone (bGH) gene have increased body weight and severe glomerulosclerosis leading to death in uremia. EXPERIMENTAL DESIGN: The aim of this study was to determine if body growth and glomerulosclerosis were mediated by different bGH regions. Amino acid substitutions in the bGH alpha-helix III were generated, and lines of transgenic mice that expressed these products were developed. Female transgenic mice carrying the native bGH gene (bGH mice), a mutated bGH gene that encodes a destabilized alpha-helix III (bGH-L121P, E126G; bGH-m11 mice), or a mutated bGH gene that encodes a perfect amphiphilic alpha-helix III (bGH-E117L, G119R, A122D; bGH-m8 mice) were examined at 2-3 months and 6-9 months of age. Body, kidney, and heart weights were measured. Urinary glucose, albumin, creatinine, and serum glucose were measured in all mice. Serum levels of insulin-like growth factor I (IGF-I) were measured in the 2-3 month group. Whole blood hemoglobin A1 was measured in some mice of the 6-9 month group. Kidney sections were examined by light and immunofluorescence microscopy. Glomerular volume was measured and related to body weight by allometry. RESULTS: The bGH-m11 mice developed glomerulosclerosis indistinguishable from that seen in bGH transgenic mice, even though they had normal body size. Glomerular growth exceeded body growth by allometry in both bGH and bGH-m11 strains. bGH-m8 mice had glomeruli of appropriate size and normal histologic appearance; however, they were dwarfs. IGF I was increased in bGH mice; they also had an increased albumin/creatinine ratio at 6-9 months. None of the mice were hyperglycemic. CONCLUSIONS: These data indicated that development of glomerulosclerosis and body growth promotion were mediated by different regions of the growth hormone molecule. The glomerular response to bGH was unique and consisted of increased size and glomerulosclerosis. PMID- 8423679 TI - Smooth muscle cell to elastic lamina connections in developing mouse aorta. Role in aortic medial organization. AB - BACKGROUND: The structural and functional intigration of smooth muscle cells and elastic laminae in the aortic media is not well established. Detailed information concerning normal ultrastructural features of the aortic media will provide a better understanding of the medial changes that occur in vascular diseases such as hypertension and aortic aneurysms. EXPERIMENTAL DESIGN: The ultrastructural development and organization of connections between smooth muscle cells and elastic laminae in the mouse aortic media were studied by light and electron microscopy. RESULTS: Early in development, the smooth muscle cells become linked to the elastic laminae by bundles of microfibrils. These microfibrils become progressively infiltrated with elastin so as to form extensions of elastin from the elastic laminae in the adult media. Each elastin extension spans obliquely from the elastic lamina to the surface of the smooth muscle cell where it attaches in a region of membrane occupied by an intracellular membrane-associated dense plaque. On the cytoplasmic face of the plaque, a contractile filament bundle penetrates and anchors in an orientation similar to that of the extracellular elastin extension. The contractile filament bundle traverses the cell obliquely and anchors in a dense plaque on the opposite side of the cell that is in turn linked to the next elastic lamina by another elastin extension. The extracellular elastin extensions and the intracellular contractile filament bundles thus form a "contractile-elastic unit," a continuous line of structures that links adjacent elastic laminae. The oblique orientation of the contractile elastic units reverses direction in successive smooth muscle cell layers in a herringbone-like pattern. Thus, tension transmitted to one elastic lamina by the smooth muscle cells on either side results in a uniform force exerted on the elastic lamina in one circumferential direction, that on the adjacent elastic laminae being in the opposite direction. CONCLUSIONS: Results from this study demonstrate the presence of smooth muscle cell to elastic lamina connections that form early in development as contractile-elastic units; basic units of aortic medial ultrastructure. The overall organization of the contractile-elastic units within the aortic media is proposed to provide a means for coordinating contractile and elastic tensions in response to mechanical stresses imposed on the vessel wall. PMID- 8423680 TI - Dr. Lucy Hughes Brown (1863-1911). A pioneer African-American physician. PMID- 8423681 TI - Portrait of a woman doctor: Love Rosa Hirschmann Gantt "Blackwellian" or "Jacobian"? PMID- 8423682 TI - Jane Bruce Guignard, M.D.: 1876-1963. PMID- 8423683 TI - Hilla Sheriff, M.D.: 1903-1988. First lady of public health. PMID- 8423684 TI - "That crazy Anne Austin". Anne Austin Young, M.D. 1892-1989. PMID- 8423685 TI - South Carolina's pioneer women doctors. Four additional trailblazers of interest. PMID- 8423686 TI - South Carolina women in medical school, 1892-1992. PMID- 8423687 TI - Psychiatrist and humanitarian Sarah Campbell Allan (1861-1954): South Carolina's first licensed woman physician. PMID- 8423688 TI - 75th anniversary of dental surgery at the Mayo Clinic. PMID- 8423690 TI - Wegener's granulomatosis associated with vasculitis of the temporal artery: report of five cases. AB - Between January 1973 and October 1991 at the Mayo Clinic, 5 of 345 patients with Wegener's granulomatosis initially had features suggestive of giant cell arteritis and subsequently were found to have biopsy-proven vasculitis of the temporal artery. All five patients were older than 60 years of age and had jaw claudication, sudden loss of vision, severe headache with or without diplopia, or polymyalgia rheumatica at the time of initial examination. The erythrocyte sedimentation rate was high at the time of onset of symptoms in four patients (and unavailable in one patient). A temporal artery biopsy specimen revealed giant cell arteritis in one patient and non-giant cell arteritis in four patients. All five patients subsequently had pulmonary and renal lesions characteristic of Wegener's granulomatosis, with typical histopathologic features on biopsy or positive cytoplasmic staining antineutrophil cytoplasmic antibodies. Thus, overlapping features of giant cell arteritis and Wegener's granulomatosis do occur in some patients. PMID- 8423689 TI - Keeping hostility in perspective: coronary heart disease and the Hostility Scale on the Minnesota Multiphasic Personality Inventory. AB - Three prospective studies from the early 1980s found that high scores on the Hostility (Ho) Scale of the Minnesota Multiphasic Personality Inventory (MMPI) were associated with an increased incidence of coronary heart disease (CHD) and mortality from CHD and other causes. In the current study, the association between the Ho score and subsequent health status was examined in a 20-year follow-up of 620 general medical patients who completed the MMPI between 1962 and 1965. Univariately, the Ho score from the MMPI was a significant factor for predicting the development of CHD, CHD-related mortality, and total mortality. When two simple risk factors for CHD--age and sex--were also considered, however, the MMPI Ho score was no longer a significant predictive factor. PMID- 8423691 TI - Pseudosclerodermatous panniculitis after irradiation: an unusual complication of megavoltage treatment of breast carcinoma. AB - An unusual edematous and indurated erythema developed in four patients with breast carcinoma 1 to 6 months after conservative surgical treatment and irradiation. The radiation therapy consisted of megavoltage x-ray photon with or without either electron beam or iridium-192 interstitial boost. Several tissue biopsy specimens revealed pronounced lymphocytic dermal and fat inflammation in conjunction with focal areas of plasma cells. The connective tissue bundles were enlarged and hyalinized. Macrophages and isolated giant cells were noted in the dermis. One biopsy specimen showed elastic tissue in giant cell cytoplasm. No mucin, fibrin, formation of cysts, or calcification was present. Lipophages and hyaline connective tissue replaced some fat lobules. The radiation-induced changes of dilated and hyalinized blood vessels, endothelial cell hyperplasia, fibrosis associated with involution of epidermal appendages, and fibroblasts were present. This combination of radiation-related and inflammatory pathologic changes is unusual and emphasizes the remarkable qualities of this rare reaction. The clinical differential diagnoses of recurrent carcinoma, cellulitis, and connective tissue disease can be excluded by reviewing the pathologic characteristics. PMID- 8423692 TI - Aortopulmonary window: factors associated with early and late success after surgical treatment. AB - Between 1953 and 1990, 19 patients, who were from 7 weeks to 27 years old, underwent surgical correction of aortopulmonary window at the Mayo Clinic. Associated cardiac anomalies were present in nine patients (47%). At operation, extracorporeal circulation was used in all except one patient. In seven patients, division and primary closure were done. In four patients, the defect was closed by direct suture through a transpulmonary approach. In eight patients, the defect was closed with a patch through a transpulmonary or transaortic approach. Four deaths (21%) occurred intraoperatively or immediately postoperatively. All four patients had undergone division of the aortopulmonary window before 1962, and three of them had a pulmonary vascular resistance (Rp) that was 7.3 U.m2 or more and a ratio of Rp to systemic vascular resistance (Rp/Rs) that exceeded 0.6. One patient with an Rp of 11.8 U.m2 and an Rp/Rs of 0.72 died 16 years postoperatively. Statistical analysis of risk factors indicated that early year of operation (P = 0.022), division of the aortopulmonary window versus transaortic or transpulmonary closure (P = 0.009), and a high Rp/Rs (P = 0.021) were significantly associated with perioperative death. All patients with a preoperative Rp of 8 U.m2 or less, an Rp/Rs of less than 0.4, and no major associated cardiac anomalies were in functional class I (New York Heart Association) postoperatively. Our study confirms that infants with aortopulmonary window should undergo operation early, before irreversible pulmonary vascular changes have developed. Patients with an Rp/Rs that exceeds 0.4 should be thoroughly assessed to determine their operability. PMID- 8423693 TI - Use of intracoronary ultrasound imaging for assessing left main coronary artery disease. AB - Coronary angiography has many limitations for the assessment of coronary artery disease. Intracoronary ultrasound imaging may overcome some of these limitations by providing direct visualization of the luminal area and plaque morphologic features. Although the size of the currently available intracoronary ultrasound catheters precludes their use in many diseased coronary vessels, lesions in the relatively large vessels, such as the left main coronary artery, can be readily assessed. Intracoronary ultrasound imaging was performed in five patients in whom the status of the left main coronary artery was unclear after conventional coronary angiography. Qualitative assessment of atherosclerotic involvement and quantitative analysis of the absolute luminal area and the percentage of area of stenosis were performed. No complications were associated with the intracoronary ultrasound procedure. In all five patients, the ultrasound studies provided additional information on which a clinical decision could be made. Intracoronary ultrasound imaging is useful for assessing disease of the left main coronary artery in selected patients in whom current angiographic techniques have provided equivocal results. PMID- 8423694 TI - Hypertension and dyslipidemia in patients with insulinoma. AB - Hyperinsulinemia is alleged to contribute to the pathogenesis of hypertension and dyslipidemia (hypertriglyceridemia) in the setting of insulin resistance. To assess the association among hyperinsulinemia, hypertension, and hypertriglyceridemia in the absence of insulin resistance, we determined their prevalence in a large cohort of patients with insulinoma (N = 250). In this retrospective case-control study, patients with insulinoma were matched by age, gender, race, and year of operation with 217 control patients admitted to the hospital for elective cholecystectomy. Mean preoperative blood pressure measurements were compared between study patients and control patients. In addition, age-, gender-, and race-specific percentiles of blood pressure were compared with data from the National Health and Nutrition Examination Survey I, and those of triglycerides (N = 65) and cholesterol (N = 70) were compared with Mayo Clinic normal reference data. The study group consisted of 105 men and 145 women; the median age was 41 years (range, 8 to 82). The median duration of symptoms before operation was 1.9 years (range, 0.05 to 40 years). After adjustment for body mass index, no statistically significant differences in systolic and diastolic blood pressure were noted between patients with insulinoma and matched control patients (131 +/- 19 versus 128 +/- 18 mm Hg and 81 +/- 11 versus 79 +/- 9 mm Hg, respectively). No relationship was observed between duration of hyperinsulinemia (as long as 40 years) and blood pressure. The age- and gender-specific percentiles of systolic and diastolic blood pressure of the patients with insulinoma did not differ from the age- and gender-specific percentiles for the general white population.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423695 TI - Magnetic resonance imaging of the liver: current clinical applications. AB - Magnetic resonance imaging provides excellent anatomic depiction of the liver and important information about focal and diffuse diseases that affect this organ. In this report, the current clinical applications of magnetic resonance imaging of the liver in patients with known or suspected hepatic metastatic lesions, hepatocellular carcinoma, cavernous hemangiomas, focal fatty infiltration, focal nodular hyperplasia, hepatic adenoma, regenerative nodules in hepatic cirrhosis, or iron overload are reviewed, and the limitations of the technique are discussed. PMID- 8423696 TI - Magnetic resonance angiography and detection of renal artery stenosis in a patient with impaired renal function. AB - Diagnosing renovascular disease in patients with renal insufficiency has challenged physicians for many years. Although contrast angiography is the "gold standard," it is associated with major risks in patients with preexisting renal failure. Other noninvasive tests have not proved to have sufficient sensitivity and specificity to supplant angiography. Developments in magnetic resonance (MR) angiographic technology, however, now enable physicians to assess the vasculature noninvasively and without use of potentially nephrotoxic agents. Herein we describe a patient with hypertension and renal failure in whom MR angiography proved to be the only effective noninvasive test for diagnosing renal artery stenosis. In addition, we review the current literature on MR angiography for renovascular disease. In the setting of renal impairment, MR angiography may be useful in screening patients for renovascular disease. More studies are needed in order to refine MR angiographic techniques and, ultimately, to determine specific situations in which MR angiography may be useful. PMID- 8423697 TI - Psychosocial issues in oncologic practice. AB - Impressive gains in the survival of some patients with malignant diseases have primarily reflected the availability of multimodality programs for selected pediatric neoplasms and germ cell tumors and for subsets of patients with regional breast cancer, colorectal cancer, and small-cell lung cancer. Most patients with advanced solid tumors, however, will die of their disease. Sophisticated psychosocial investigations of patients with advanced cancer have targeted several areas in which clinicians can positively influence quality of life. Families often "cascade through an avalanche" of emotional upheavals as patients struggle with the sequelae of their illness. After a patient dies, clinicians should be familiar with some generally recognized patterns of behavior that are indicative of a normal mourning process. This knowledge may help clinicians be aware of situations that might necessitate intervention of other professionals, either medical or pastoral. Attention to psychosocial events is an integral part of a comprehensive oncologic program to facilitate patients and families to live in an atmosphere of peace and dignity. PMID- 8423698 TI - The etiology of lung cancer. AB - The association between tobacco smoking and lung cancer has been noted for more than 50 years and continues to dominate the etiologic milieu of this malignant disease. Other agents, many discovered in the occupational setting, have also been substantiated as lung carcinogens. Inherent predisposition to the disease has long been suspected, and recent investigations suggest several potential mechanisms and a possible mode of inheritance. Considerable progress has been made in deciphiring the molecular defects present in lung cancer cells. These recent findings have been incorporated into two well-known models of lung carcinogenesis. As the details of the carcinogenic process are unraveled, one goal is to identify intermediate (preneoplastic) markers of exposure and inherent predisposition that will help assess the risk of lung cancer for individuals as well as for groups. PMID- 8423699 TI - Occupational lung cancer. AB - Exposure to certain industrial agents has been thought to have carcinogenic potential, both for employees who work closely with such agents and for the general population that comes in contact with them. Although case reports, laboratory studies, and epidemiologic analyses help to determine the carcinogenicity of implicated agents, each of these types of investigation has limitations and deficiencies in distinguishing causal from noncausal associations. Asbestos has been linked with bronchogenic carcinoma, but several controversial factors--the degree of risk relative to exposure dose, the synergistic effect of cocarcinogens, and the question of existence of a threshold dose--complicate the understanding of the magnitude of the risk for exposed persons. Several other physical and chemical agents (such as chromium, nickel, and radon) have also been associated with an increased incidence of lung cancer in epidemiologic and animal studies. As with asbestos, the specific type of the agent and exposure conditions are important in determining the degree of carcinogenicity. In studies of exposure to man-made mineral fibers, formaldehyde, and silica, the findings have been inconsistent. Because the degree of health hazard attributable to asbestos and other known and suspected lung carcinogens is controversial, a wide range of opinions exists about the importance of occupational exposures to the overall incidence of lung cancers. Nevertheless, attempting to prevent lung cancers by minimizing or eliminating exposure to carcinogens is preferable to treating existent cases. PMID- 8423700 TI - Dr. Jennie Kidd Trout--pioneer Canadian physician. PMID- 8423701 TI - Keeping conflicting findings in perspective: the case of hostility and health. PMID- 8423702 TI - Vasculitis masquerading as vasculitis. PMID- 8423703 TI - 3-(Phenylamino)alanine--a link between eosinophilia-myalgia syndrome and toxic oil syndrome? PMID- 8423704 TI - Allergic reactions to latex. PMID- 8423705 TI - Colchicine alters lactate utilization in isolated hepatocytes of rats treated with CCl4 and ethanol. AB - Lactic acidosis has been described in patients with liver disease. Hyperlactacidaemia results from an imbalance in lactate production versus lactate utilization. It is estimated that the liver utilizes approximately 30 percent of the total lactate produced in the body under basal conditions, primarily by gluconeogenesis. The gluconeogenesis from lactate 10 mM and lactacidaemia were determined in order to investigate the effects of CCl4+ethanol administration in liver injury and, the possible effect of colchicine in our experimental fibrosis model. The tests were determined after 15, 30 or 45 days of treatment. The results indicate that the gluconeogenesis was significantly inhibited in both CCl4+ethanol groups and CCl4+ethanol+colchicine groups. By contrast, the lactacidaemia levels were much higher in the CCl4+ethanol groups than the colchicine groups. Summarising, we have documented that hyperlactacidaemia is due to the inhibition of lactate utilization by the isolated hepatocytes in experimental cirrhosis, and that the improvement in lactacidaemia caused by colchicine is not primarily due to an increase in hepatic lactate utilization. PMID- 8423706 TI - ANP(1-28), BNP(1-32) and CNP(1-22) increase the severity of picrotoxin-kindled seizure syndrome in rats. AB - The administration of rANP(1-28) in doses of 1.0 and 2.0 nmol (but not 0.2 nmol) into the lateral cerebral ventricle (i.c.v) of rats preliminarily kindled with picrotoxin resulted in an increase of the severity of picrotoxin-kindled convulsions 24 hrs after injection of the peptide. I.c.v. injection of pBNP(1-32) also resulted in a proepileptic effect when it was applied in the same doses with a similar time course; the increased seizure severity was observed 48 hrs after injection of pBNP in a dose of 2 nmol. I.c.v. administration of CNP(1-22) in a dose of 2 nmol induced an increase in the severity of picrotoxin-kindled convulsions 24 and 48 hrs after application of the peptide. None of the peptides influenced the seizure syndrome immediately after the injections. It is presumed that the delayed proepileptic properties of the three natriuretic peptides could be caused by some of their stable fragments which accumulate during their metabolism. PMID- 8423707 TI - Minireview: Molecular genetics in affective illness. AB - Genetic transmission in manic depressive illness (MDI) has been explored in twins, adoption, association, and linkage studies. The X-linked transmission hypothesis has been tested by using several markers on chromosome X: Xg blood group, colour blindness, glucose-6-phosphate dehydrogenase (G6PD), factor IX (haemophilia B), and DNA probes such as DXS15, DXS52, F8C, ST14. The hypothesis of autosomal transmission has been tested by association studies with the O blood group located on chromosome 9, as well as linkage studies on chromosome 6 with the Human Leucocyte Antigens (HLA) haplotypes and on Chromosome 11 with DNA markers for the following genes: D2 dopamine receptor, tyrosinase, C-Harvey-Ras-A (HRAS) oncogene, insuline (ins), and tyrosine hydroxylase (TH). Although linkage studies support the hypothesis of a major locus for the transmission of MDI in the Xq27-28 region, several factors are limiting the results, and are discussed in the present review. PMID- 8423708 TI - Bidirectional changes in striatal D1-dopamine receptor density during chronic ethanol intake. AB - Results of previous studies on the effects of ethanol consumption on the properties of D1 dopamine receptors appear contradictory and inconclusive. In this study we have examined the time course of the effects of dietary ethanol on the properties of striatal D1 dopamine receptors. Chronic ethanol intake led to bidirectional changes in the maximum number (Bmax) of [3H]SCH-23390 binding to striatal D1 dopamine receptors measured 10 hrs after termination of the ethanol intake. A significant decrease (80% of control), increase (159% of control), increase (122% of control), and decrease (85% of control) after 1, 2, 3, and 4 weeks of ethanol intake respectively was observed. The bidirectional changes disappeared after 6 and 10 weeks of continued ethanol intake and the Bmax returned back to the control level. The receptor affinity (Kd), however, remained unaltered in all cases. These data suggest that the duration of ethanol exposure may be an important determinant in regulating D1 dopamine receptor density. PMID- 8423709 TI - Chronic treatment of (-)deprenyl prolongs the life span of male Fischer 344 rats. Further evidence. AB - Seventy male Fischer 344 (F-344) rats were treated with s.c. injection of ( )deprenyl (0.5 mg/kg, n = 35) or physiological saline (n = 35) 3 times a week from the age of 18 months until the time of their natural death. The fifty percent survival time was 28 months in control animals and 30 months in the deprenyl treated group. The mean survival time after the start of treatment (18 months) and after 24 months were 378.3 +/- 97.4 days (mean +/- SD) and 196.3 +/- 97.4 days, respectively, in deprenyl treated rats and 328.7 +/- 108.8 days and 146.7 +/- 108.7 days in control rats. The increases in average life expectancies caused by deprenyl treatment (15% from 18 months and 34% from 24 months) were both statistically significant (P < 0.05, two-tailed t-test). The average body weights were comparable for both groups but the variation of body weight was greater in control groups, thus excluding the possibility that the life prolonging effect of deprenyl results from reduced dietary intake. The results confirm those of two previous studies (1,2) which reported a significant life prolonging effect of deprenyl in aged rats and lend added support to the results of a study on male F-344 rats where the effect was only marginally significant (16% increase after 24 months, P = 0.048 by one-tailed t test) (2). PMID- 8423710 TI - A possible mechanism of antidepressant activity of beta-amyrin palmitate isolated from Lobelia inflata leaves in the forced swimming test. AB - A mechanism of antidepressant activity of beta-amyrin palmitate was studied using the forced swimming method in mice. Beta-amyrin palmitate (10 mg/kg) reduced the increase in the duration of immobility induced by tetrabenazine (100 and 200 mg/kg), but showed no effect on that in mice treated with alpha-methyl-para tyrosine (500 mg/kg). Beta-amyrin palmitate (5 and 10 mg/kg) decreased the duration of immobility in mice treated with desipramine plus 6-hydroxy-dopamine (50 micrograms/mouse), but did not affect that induced by nomifensine plus 6 hydroxydopamine. The decreased immobility produced by desipramine (15 mg/kg) was not affected by beta-amyrin palmitate. A study of norepinephrine release in mouse brain synaptosomes indicated that beta-amyrin palmitate caused a release of [3H]norepinephrine. The results of the present study suggest that beta-amyrin palmitate might release norepinephrine from newly synthesized pools, and thus, it might activate noradrenergic activity. PMID- 8423711 TI - Human brain cortex and platelet serotonin2 receptor binding properties and their regulation by endogenous serotonin. AB - In patients with affective disorder and suicidality up-regulation of the serotonin2 receptor has been observed in brain and on platelets. Although the pharmacological profiles of the receptor in brain synaptosomes and platelet membranes are similar, it is a matter of debate whether the platelet serotonin2 receptor reflects the characteristics of the receptor in the brain and whether serotonin regulates the receptor activity. To answer these questions we measured in healthy human subjects the platelet serotonin2 receptor activity and blood serotonin concentrations. In an attempt to find whether the serotonin2 receptor activity in brain cortex synaptosomes and on platelets is similarly expressed we investigated the receptor's binding characteristics in neurosurgical patients. The results suggest that in men and women increased platelet serotonin concentrations correlate with a decrease in platelet membrane serotonin2 receptor affinity. The affinities of the brain cortex synaptosomal and platelet membrane serotonin2 receptor correlate intra-individually. These data suggest that the platelet serotonin2 receptor affinity appears to be regulated at the cellular level by blood serotonin and that the binding characteristics of the serotonin2 receptor in brain cortex synaptosomes corresponds to that on platelets. The latter finding supports the hypothesis of the platelet as a model for neuronal function. PMID- 8423712 TI - Malondialdehyde modified proteins and their antibodies in the plasma of control and streptozotocin induced diabetic rats. AB - One of the possible mechanisms of diabetes-related tissue damage is modification of various proteins via lipid peroxidation byproducts such as malondialdehyde (MDA). To determine the extent of MDA derivatization of plasma proteins, Western blots were carried out using anti-MDA antisera to study plasma proteins in control and streptozotocin (STZ)-induced diabetic rats. Since MDA can modify proteins and may alter or enhance their antigenicity, we screened plasma samples for anti-MDA antibodies using enzyme-linked immunosorbent assay (ELISA), and confirmed antibody specificity by inhibition ELISA. Circulating immune complexes containing MDA were also assayed. This study is the first demonstration of the existence in plasma of MDA-modified proteins with a molecular weight of approximately 100 Kd. Both control and diabetic rats have similar concentrations of plasma anti-MDA antibodies and circulating immune complexes. These results do not support the notion that diabetes alters the immune response to MDA modified proteins. Whether MDA modification of proteins participate in immunological processes that lead to tissue injury remains to be demonstrated. PMID- 8423713 TI - In vivo quantification of brain metabolites by 1H-MRS using water as an internal standard. AB - The reliability of absolute quantification of average metabolite concentrations in the human brain in vivo by 1H-MRS using the fully relaxed water signal as an internal standard was tested in a number of in vitro as well as in vivo measurements. The experiments were carried out on a SIEMENS HELICON SP 63/84 wholebody MR-scanner operating at 1.5 T using a STEAM sequence. In vitro studies indicate a very high correlation between metabolite signals (area under peaks) and concentration, R = 0.99 as well as between metabolite signals and the volume of the selected voxel, R = 1.00. The error in quantification of N-acetyl aspartate (NAA) concentration was about 1-2 mM (6-12%). Also in vivo a good linearity between water signal and selected voxel size was seen. The same was true for the studied metabolites, N-acetyl aspartate (NAA), creatine/phosphocreatine (Cr/PCr), and choline (Cho). Calculated average concentrations of NAA, Cr/PCr, and Cho in the occipital lobe of the brain in five healthy volunteers were (mean +/- 1 SD) 11.6 +/- 1.3 mM, 7.6 +/- 1.4 mM, and 1.7 +/- 0.5 mM. The results indicate that the method presented offers reasonable estimation of metabolite concentrations in the brain in vivo and therefore is useful in clinical research. PMID- 8423714 TI - Altered muscle metabolism shown by magnetic resonance spectroscopy in sickle cell disease with leg ulcers. AB - We performed 31P magnetic resonance spectroscopy of gastrocnemius muscle at rest in 7 normal volunteers and 12 patients with sickle cell disease (7 with leg ulcers and 5 without leg ulcers but with painful crises). We measured intracellular pH and ratios of Pi to ATP, PCr to ATP and PCr to Pi (Pi = inorganic phosphate, ATP = adenosine triphosphate, and PCr = phosphocreatine). Magnetic resonance arteriograms were also performed. Significant differences were found for PCr/Pi ratios between normals and sickle cell disease patients with leg ulcers (p < 0.008). Magnetic resonance arteriograms were normal in volunteers and patients with sickle cell disease. The altered high energy phosphate metabolism in sickle cell disease with leg ulcers is consistent with muscle ischemia or hypoxia. PMID- 8423715 TI - Ni-DTPA doped agarose gel--a phantom material for Gd-DTPA enhancement measurements. AB - In order to study the relationship between the concentration of Gd-DTPA in tissue, and the resulting changes in relaxation times and signal intensity, a phantom material that has similar relaxation times to tissue and that can be doped with Gd-DTPA is required. The "tissue-equivalent" material should not contain Gd; nor should it alter the relaxivities of Gd-DTPA from their values in aqueous solution (R1 = 4.5 sec-1 mM-1; R2 = 5.5 sec-1 mM-1 at 1.5 T). Conventional materials, based on CuSO4-, MnCl2-, or GdCl3/LaCl3-agarose mixtures, are not suitable, since Gd is displaced from the Gd-DTPA chelate. The new material, consisting of Ni-DTPA dissolved in agarose, is easy to prepare and does not interact with Gd-DTPA. Its relaxation times are stable; relaxivity R1 was within 4% of its aqueous value over 109 days. T1s have low dependence on temperature (0.2-1.0%/degrees C at 21 degrees C) and on field strength, allowing the material to be used as a relaxation time standard for quality assurance. Equations giving the concentration of Ni-DTPA and agarose to produce a required T1 and T2 are provided. PMID- 8423716 TI - Spinal intramedullary granulocytic sarcoma: magnetic resonance imaging. AB - A rare case of spinal intramedullary granulocytic sarcoma (GS) in a patient with acute myelogenous leukemia is described along with its magnetic resonance (MR) finding. The mass shows isointense signal on T1-weighted images, and slightly higher intensity on T2-weighted images. Contrast enhancement with gadolinium (Gd) DTPA in this case differs from the other report in which the tumor was not enhanced. As MRI becomes the first choice in the evaluation of spinal tumors, high index of suspicion of GS with familiarity with its MR finding in leukemic patients may obviate the need for surgical intervention, since the tumor is sensitive to both radiation and chemotherapy. PMID- 8423717 TI - Effectiveness of the Short TI Inversion Recovery (STIR) sequence in MR imaging of intramedullary spinal lesions. AB - A Short TI Inversion Recovery (STIR) sequence with spin-echo data collection was compared to a conventional cardiac gated long TR spin-echo (SE) sequence for detecting intramedullary signal abnormalities. The cervical (n = 48), cervico thoracic (n = 1), and thoraco-lumbar (n = 18) spinal cord was imaged in a sagittal plane with a 0.5 T (n = 61) or 1.5 T (n = 6) MRI unit in 67 patients with clinical evidence of myelopathy of different etiologies (e.g., multiple sclerosis, trauma, herniated intervertebral disk, spondylosis, etc.). In all patients, ungated double or quadruple echo STIR images (TR 1000-1400 msec, TI 100 msec, TE 30-60 or 30-60-90-120 msec) were compared with cardiac gated long TR (1400-2100 msec), double echo (30-100, 50-100, or 50-150 msec) SE images with first order flow compensation for the second echo. Although STIR images appeared "noisier" than long TR SE images, they showed fewer ghost artifacts. In 55 patients, single or multiple, focal or diffuse, hyperintense areas within the spinal cord were observed on both long TR SE and (magnitude reconstructed) STIR images. Lesion conspicuity was better on the STIR images in 25 patients, better on the SE images in 14 patients, and equal in 16. STIR sequence provides a valuable alternative to gated long TR SE sequence for the MRI investigation of intramedullary spinal lesions. PMID- 8423718 TI - Low resolution spin echo: a simple timesaving technique for MRI liver exams. AB - MR evaluation of the liver at mid-field strength requires relatively lengthy T2 weighted sequences (T2W) for differentiation of benign cavernous hemangiomas from malignant lesions. A short duration T2W, which maintains standard signal-to-noise ratio and also contrast relationships, can be easily implemented by increasing the pixel size in the phase-encoding direction (thus reducing spatial resolution) and proportionally decreasing the number of phase-encoding steps in the matrix (thus reducing acquisition time). Blinded interpretations of a quick (4 min), low resolution (3.4 mm x 1.7 mm pixel) T2W sequence (matrix 64 x 256, FOV 21.7 cm phase x 43.5 cm frequency) were compared to the 17 min standard resolution (1.7 mm x 1.7 mm) T2W sequence (256 x 256 matrix, FOV 43 x 43 cm) in 25 patients suspected of having liver metastasis. Lesions felt to be cavernous hemangiomas showed a 100% (24/24) agreement for interpreter "A" and 96% (22/23) agreement for interpreter "B" when 4 min low resolution T2W was compared to the standard 17 min sequence. Sensitivity (for all types of lesions) of the low resolution T2W sequence ranged from 100% (31/31) for interpreter "A" to 80% (28/35) for interpreter "B." Missed lesions (interpreter "B") were either partially obscured by excessive fat (wrap around) (N = 4), less than 1 cm in size (N = 2), or degraded by motion artifact (N = 1). Thus in many situations low resolution T2 may provide a substantial timesaving alternative to standard T2W particularly where T2W is used primarily for lesion classification in normal sized patients. PMID- 8423719 TI - Coronary artery imaging in multiple 1-sec breath holds. AB - The act of breathing results in considerable bulk cardiac motion, thus breath holding is usually required to image the coronary arteries. Normally, breath hold periods of about 15 sec are required, which can be difficult to sustain over several minutes. We introduce a breath holding strategy that only requires a 1 sec breath hold in every 4 sec. A set of cross-sectional images is acquired in approximately 10 min, and 3-dimensional renderings of the coronary tree are produced from these. PMID- 8423720 TI - Magnetic resonance imaging of female pelvic masses and local recurrent tumors at an ultra low (0.02 T) magnetic field: correlation with computed tomography. AB - Pelvic MR (41 patients) and CT (36 patients) examinations were performed on 14 females with a primary pelvic complaint, and on 28 females with a suspicion of local recurrent disease of gynecologic malignant tumor. Benign cystic tumors were found in eight patients, five patients had endometriomas, one had a lymphoma, and one had a small androblastoma. Ten local recurrent tumors were confirmed histopathologically or cytologically. All cysts, one endometrioma, the lymphoma, and six recurrent tumors were detected on images obtained by our ultra low field magnetic unit. The smallest cyst detected was 16 mm in diameter. Small scattered implants of endometriosis were not discerned. The appearance of the tumors did not differ essentially from those described at high magnetic fields. Physical examination detected all 10 recurrent tumors, CT detected 8 of them, and MRI 6 out of 9 cases. The sensitivities of physical examination, CT and MRI to find recurrent diseases were 100%, 80%, and 67%, respectively. Corresponding specificities were 93%, 67%, and 80%. The results indicate that physical examination is most important in recurrent diseases. CT is the basic method for imaging malignant pelvic tumors. MR imaging at 0.02 T magnetic field provides a convenient and inexpensive method for more specific information, if CT findings are equivocal. MRI at 0.02 T is also accurate in detecting benign pelvic masses, but the findings are not very specific. PMID- 8423721 TI - Giant extra-adrenal pheochromocytoma: magnetic resonance imaging with gadolinium DTPA enhancement. AB - A case report of a 35-yr-old male presenting with moderate hypertension and a giant extra-adrenal pheochromocytoma is discussed. The patient was treated by chemotherapy and surgery with regression of the lesion and clinical relief of symptoms. Initial magnetic resonance imaging (MRI) with gadolinium-DTPA enhancement revealed a large complex pelvic mass. Follow-up examination following chemotherapy three months later demonstrated partial regression. The MRI findings and the pattern of gadolinium-DTPA enhancement are discussed. PMID- 8423722 TI - Measurement of total and unilateral renal blood flow by oblique-angle velocity encoded 2D-cine magnetic resonance angiography. AB - Two independent measurements of total renal blood flow (RBF) were made in healthy human subjects (n = 14, mean age 30 yr) by CINE phase-contrast magnetic resonance angiography. RBF, measured by summing the flows measured in the right and left renal arteries, was 1152 +/- 44 ml/min (mean +/- SE). RBF, measured from the difference between supra- and infrarenal abdominal aorta flow, was 1109 +/- 68 ml/min. Regression analysis of the comparison of these two different RBF calculations yielded a correlation coefficient of 0.72 at a p < .05 level of significance. Based on other studies of RBF in normal subjects by para aminohippuric acid (PAH) clearance, the expected RBF in this subject group was 1211 +/- 62 ml/min. The results indicate that noninvasive measurement of RBF is possible using phase-contrast magnetic resonance methods. PMID- 8423723 TI - MRI mapping of postreconstructive edema following femoropopliteal bypass surgery. AB - We have used T1 MR images to map the distribution of water contributing to the edema which follows femoropopliteal bypass surgery. Spin-echo images and true parametric T1 images were made at the same time. The spin-echo images were used to identify the tissue anatomy. The extra fluid contributing to the edema distributes in two phases: a volume equivalent to 5% of the leg volume is distributed throughout the leg tissue, while the excess fluid collects in a localized annulus lying adjacent to the fascia. The T1 in the annulus can be as high as 4 sec, indicating build up of free fluid in this region. Although most of this free fluid lies in the subcutaneous fat, there is a component which lies underneath the fascia in the muscle compartment. PMID- 8423724 TI - Magnetization transfer imaging of the abdomen at 0.1 T: detection of hepatic neoplasms. AB - Magnetization transfer (MT) techniques have been proposed as a method of increasing contrast in MR images. To evaluate the feasibility of MT imaging of the abdomen at 0.1 T and to assess the clinical utility of this technique, the authors studied tissue contrast with a gradient-echo pulse sequence and an MT sequence in four normal volunteers, and in 17 patients with known primary or secondary neoplasms of the liver. The MT technique increased contrast between the liver and other tissues such as spleen, skeletal muscle and subcutaneous fat. The technique also produced increased contrast between hepatic tumors and normal liver parenchyma in gradient-echo images. PMID- 8423725 TI - Evaluation of renal masses with contrast-enhanced rapid acquisition spin echo MR imaging. AB - To assess the use of dynamic contrast-enhanced rapid acquisition spin-echo (RASE) imaging for the detection and characterization of renal masses, we evaluated 18 patients with CT evidence of 67 renal masses. The masses included 58 simple cysts, 8 solid neoplasms, and 1 pseudotumor. Patients were examined with standard spin-echo (SE) pulse sequences including pre- and postcontrast T1-weighted images and noncontrast T2-weighted images as well as pre- and postcontrast RASE images. Each pulse sequence was reviewed individually and in a group with other pulse sequences by two blinded observers. The performance of contrast-enhanced RASE imaging either alone or in combination with a T2-weighted SE sequence resulted in improved diagnostic accuracy compared with unenhanced conventional SE and RASE sequences. The contrast-enhanced RASE sequence outperformed the contrast-enhanced T1-weighted SE sequence for one observer; similar diagnostic accuracy was achieved with the two examinations by the other observer. Combining the T2 weighted sequence with contrast-enhanced RASE images did not increase lesion detection but did increase the observers' confidence in making the diagnosis. Contrast-enhanced MR imaging resulted in significant improvement in the detection and characterization of renal lesions compared to unenhanced MR imaging. PMID- 8423726 TI - Design and optimization of a breast coil for magnetic resonance imaging. AB - Mathematical methods were implemented to optimize the configuration of specially designed breast coils leading to substantial improvements in magnetic field homogeneity. Multiple-turn shaped coils were constructed according to mathematically determined parameters derived from B1 field optimization calculations. Significant improvements in coil homogeneity were obtained without overcompromising quality factors and signal-to-noise ratios (S/N). Coil performance was verified using both phantom and breast images, revealing improved coil performance particularly in the region adjacent to the chest wall. These new coils are easily constructed and can be designed to accommodate various breast sizes. They can be utilized in the transmit/receive or receive only mode and can be doubly tuned for multinuclear spectroscopic studies. Moreover, nothing precludes their use to image both breasts simultaneously. The results demonstrate the ease of our approach and suggests that these methods can be feasibly applied to other surface coil designs. PMID- 8423727 TI - An adjustable RF coil loading device. AB - This paper describes an adjustable loading device that can substitute for the tissue losses of various sized patients in a whole-body MR imager. It resembles a lowpass birdcage resonator but with resistors replacing the capacitors on the cylindrical surface. Power dissipated is a monotonic function of the total surface conductance of the loader. The loader can be used in conjunction with a low loss water filled phantom to measure signal-to-noise ratios (SNR). The impact on measured SNR due to screening by the loader and changing the phantom size or composition are also briefly discussed. PMID- 8423728 TI - Implementation and optimization by the simplex method of a 3D double echo sequence in steady-state free precession. AB - Under steady-state conditions, the resulting echoes have very complex T1 and T2 relationships. Many authors exploited these echoes in different sequences to produce either T1- or T2-weighted images. The simultaneous acquisition of two echoes in a single sequence provides two images of clearly different contrasts. We implemented such a sequence, in a 3D-acquisition mode, combining the advantages of thin and contiguous slices with those of a multi-echo sequence. The contrast of the images was correlated with theoretical results, derived from Bloch equations. In order to estimate the acquisition parameters (alpha, TE, TR) to obtain an optimal T1- or T2-contrast between two tissues, a computer simulation of these equations was used in conjunction with the simplex method. The results show that this sequence improves the clinical efficiency of MRI, particularly in neurological and articular disease. PMID- 8423729 TI - MRI: stability of three supervised segmentation techniques. AB - Supervised segmentation methods from three families of pattern recognition techniques were used to segment multispectral MRI data. Studied were the maximum likelihood method (MLM), k-nearest neighbors (k-NN), and a back-propagation artificial neural net (ANN). Performance was measured in terms of execution speed, and stability for the selection of training data, namely, region of interest (ROI) selection, and interslice and interpatient classifications. MLM proved to have the smallest execution times, but demonstrated the least stability. k-NN showed the best stability for training data selection. To evaluate the segmentation techniques, multispectral images were used of normal volunteers and patients with gliomas, the latter with and without MR contrast material. All measures applied indicated that k-NN provides the best results. PMID- 8423730 TI - Breast-feeding support program is now in place. PMID- 8423731 TI - Breast-feeding support program is now in place. PMID- 8423732 TI - COBRA and patient dumping. PMID- 8423733 TI - Cognitive and behavioral strategies to reduce children's pain. PMID- 8423734 TI - Chemical assessment in maternity care. PMID- 8423735 TI - Safe transport of technology-dependent children. PMID- 8423736 TI - Assessing and promoting positive parenting in adolescent mothers. PMID- 8423737 TI - Physiologic second-stage labor. PMID- 8423738 TI - Patient education standards. PMID- 8423739 TI - Exploratory data analysis. Part II: Steps in examining data. PMID- 8423740 TI - Ritodrine, reexamined. PMID- 8423741 TI - Managed care: what does it mean? PMID- 8423742 TI - Why do I have to work so hard? PMID- 8423743 TI - Simultaneous measurement of heart rate and body motion to quantitate physical activity. AB - None of the various methods used to measure habitual physical activity over days, weeks, or years in the general population have yet proven entirely satisfactory. A major problem is that no "gold standard" exists for the validation of various questionnaires, logs, or diaries that can be used in large sample population studies. Attempts have been made to accurately measure the activity profile by using heart rate or various motion sensors or accelerometers, but each approach has had significant limitations. The availability of new solid state recording techniques and computer-based analytic and display procedures now makes it possible to simultaneously record heart rate and body movement continuously for days and to combine the analysis of these data using customized software. Preliminary evaluation of this concept of simultaneous recording and analysis of heart rate and body motion via movement sensors on an arm and leg were conducted in 19 men. Subjects performed a variety of exercises in the laboratory during which heart rate, leg motion, arm motion, and oxygen uptake were recorded. Various issues regarding the prediction of energy expenditure from heart rate and body movement independently and in combination were evaluated. The results demonstrate that the accuracy of estimating oxygen uptake during a wide range of activities is improved when individualized heart rate--oxygen uptake regressions are used and heart rate and body movement are analyzed simultaneously rather that separately. PMID- 8423744 TI - Electroencephalographic measures of attentional patterns prior to the golf putt. AB - The purpose of this investigation was to determine the attentional focus patterns associated with golf putting performance. Highly skilled golfers (N = 34) were assessed using electroencephalographic (EEG) measures of the motor and temporal cortices during the 3 s prior to the golf putt. Players completed 40, 12-ft putts and performance was measured in cm error from the hole. Three measures of EEG were analyzed: slow shift, 40 Hz, and relative power spectrum; representing readiness to respond, focused arousal, and general cortical activity, respectively. All three EEG measures suggested a decrease in left hemisphere, motor cortex activity as the player prepared to putt. Relative power measures also showed significant increases in right hemisphere activity in both the motor and temporal cortices. During the last second preceding the putt, increased right hemisphere alpha activity correlated with and predicted less error. Hemispheric differentiation was also reduced as subjects prepared to putt and few, but important, differences existed between the motor and temporal cortices. An important distinction occurred in the alpha band. In the motor cortex left hemisphere alpha increased significantly over time while in the temporal cortex, right hemisphere alpha increased as subjects approached stroke initiation. Differences that existed between the attentional patterns from the present study and past sport studies may relate to the use of one versus two hands to initiate the response. PMID- 8423745 TI - Effect of time zone and game time changes on team performance: National Football League. AB - To determine the effect of time zone and game time changes on NFL team performance, win-loss records from 1978-1987 were analyzed. Twenty-seven NFL teams were grouped by time zone and possible anti-jet lag adjustments. Among all intra-time zone rivals, home teams won 56.6%, away teams won 43.8%, for a home vs away winning percentage change of -12.8% (P < 0.001). West teams (N = 5) displayed fluctuations in home vs away team performance in association with trans meridian travel. The change in winning percentage was found to be 0.0% vs West teams, -14.1% vs Central teams (N = 8) (P < 0.05), -16.3% vs East (N = 14) (P < 0.05) for West teams (N = 4) flying about 42 h pregame and +2.3% vs East for the one West team advancing practices 3-4 h to match East coast game time in addition to 48 h pregame flights. For night games within the same time zone, home vs away team winning percentage changed -23.8% (P < 0.01). West teams displayed uniformly high home winning percentages (75.0% and 68.4%) when playing Central and East teams, respectively, with little or no fall in away winning percentages (67.7% and 68.8%). For day games, a 3-h phase advance may decrease West coast team performance. In one small subset, anti-jet lag adjustments appeared to eliminate the expected decrement in performance. For night games, West coast teams, whether home or away, appear to be at a distinct advantage over East and Central teams. PMID- 8423746 TI - Effect of training on the relationship between maximal and submaximal strength. AB - The purpose of this study was to evaluate the validity of a dynamic seven to 10 repetition maximum (7-10 RM) test to estimate maximal knee extension strength (1 RM) in untrained and trained subjects. Thirty-three men and 25 women (25 +/- 5 yr) were randomly assigned to a group that trained two or three times.wk-1 for 18 wk (N = 47) or a control group (N = 11). Training included one set of 7-10 repetitions to volitional fatigue on a Nautilus knee extension machine. Prior to (T1) and after training (T2) dynamic strength was evaluated by 1-RM and 7-10 RM tests. The 7-10 RM test consisted of one set of variable resistance knee extension exercise performed to volitional fatigue with a weight that allowed 7 10 repetitions. The training group improved their 1-RM and 7-10 RM strength (by 31.7 and 51.4%, respectively) (P < or = 0.01) while the control group did not change. Training increased relative 7-10 RM strength (68.4% of 1-RM at T1 and 79.1% of 1-RM at T2) (P < or = 0.01). The relationship between the 7-10 RM weight and 1-RM at T1 was linear: 1-RM = 1.554 (7-10 RM weight)-5.181; R2 = 0.89; SEE = 9.3 kg. Application of this equation following training resulted in a systematic overprediction (p < or = 0.01) of 1-RM (21.2 kg) in trained subjects.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423747 TI - Sources of variation in oxygen consumption during a stepping task. AB - Factors that determine oxygen uptake during stepping exercise (modified Canadian Aerobic Fitness Test, CAFT) were examined in 66 women and 55 men. Subjects ranged in age from 15 to 67 yr, VO2max from 22.5 to 76.9, leg length from 75.0 to 101.0 cm, and body mass from 48.4 to 107.7 kg. In accordance with the modified CAFT protocol, subjects stepped at cadences determined by their age and heart rate response. The oxygen demand of stepping at each cadence was measured on two occasions. A paired t-test revealed no significant (P > 0.05) difference in oxygen demand between the repeats and the intraclass correlation coefficient was 0.51. The coefficient of variation of oxygen uptake (ml O2.kg-1.min-1) at a given stepping cadence averaged 10.4%. Regression analysis indicated that little variation in oxygen demand could be explained by simple descriptors of the subjects. Age was weakly related to oxygen demand at a given level (males r = 0.26, females r = 0.58). Stepwise multiple regression of the oxygen uptake at selected stepping cadences on possible independent variables confirmed that little of the variation could be explained by age, aerobic fitness, leg length, or adiposity (maximum R2 = 0.34). We conclude that variation in the oxygen demand of a stepping task can account for a large portion of the error in predicting VO2max from a submaximal stepping test. Our ability to predict the oxygen demand of stepping from subject characteristics is limited. PMID- 8423748 TI - Anthropometric measurements and estimating body composition in ballet dancers. AB - The purpose of this study was to develop and cross-validate an equation for estimating fat-free mass (FFM) in female ballet dancers. One hundred twelve, 11- to 25-yr old, female dancers had FFM measured by total body electrical conductivity (TOBEC) and anthropometrics, including skinfold and circumference measurements. The regression equation that best estimated FFM in the dancers was FFM = 0.73 x body weight (kg) + 3.0, (R2 = 88%, SEE = 1.5 kg, P < 0.001). This equation was then cross-validated on a separate group of 23 female dancers who also had FFM measured by TOBEC. FFM estimated by this equation correlated with FFM measured by TOBEC (r = 0.94, SEE = 0.9 kg), and the difference in the FFM values using the two methods (the equation and TOBEC) did not change with the size of the FFM of the dancers. FFM in accomplished female ballet dancers can be best estimated from body weight alone. This is related to the homogeneity of body size and body composition in female ballet dancers at this level. PMID- 8423749 TI - A comparison of methods to predict minimal weight in high school wrestlers. AB - The purpose of this study was to compare the accuracy of body fat determinations and subsequent calculation of minimal weight (MW) by dual energy x-ray absorptiometry (DEXA), bioelectrical impedance (BIA), near-infrared photospectometry (NIR), and anthropometry (LOHMAN). Necessitated by mandatory state minimal weight testing, the methods were cross-validated on 95 Wisconsin high school wrestlers (mean +/- SD; age: 15.1 +/- 1.2 yr, height: 170.4 +/- 7.1 cm, weight: 63.4 +/- 9.8 kg). MW, defined as fat-free body/0.93, determined by hydrostatic weighing (HW) and residual volume via O2 dilution, served as the criterion. The validity of the four selected MW predictions were evaluated against HW by examining mean differences (MD), standard deviation differences (SDD), correlations (r), standard error of estimate (SEE), and total errors (TE). Statistically significant differences were shown between the methods and the criterion by t-tests; however, these were clinically small in Lohman (0.6 kg) and BIA (0.9 kg). TE ranged from 2.25 kg (Lohman) to 6.03 kg (NIR). The results indicated that Lohman skinfold equation provided the most accurate prediction of MW, demonstrating the highest correlation (0.972), lowest MD (0.6 kg), lowest SEE (2.12 kg), and lowest TE (2.25 kg) of the methods evaluated. PMID- 8423750 TI - Metabolic response to graded downhill walking. AB - Compared with level walking or running, progressive downhill walking or running requires a decreasing energy cost to some minimum where the cost again increases with further decrements in grade. Margaria estimated this minimum occurs at a -9% grade. In this study an attempt was made to more precisely track the energy cost curve in progressive downhill treadmill walking. Ten men, mean age 22.0 +/- 2.5 yr, volunteered as subjects. After VO2max determinations the subjects attended two downhill walking sessions. Each subject performed 14 randomly ordered walking bouts of 6 min in duration, at speeds of 90 and 105 m.min-1. The grades used were 0, -3, -6, -9, -12, -15, and -18%. Gas exchange measurements were obtained by open circuit spirometry during each work bout. Heart rate was monitored continuously and the stride frequency was counted by direct observation during each walking bout. Net VO2 values decreased with decrements in grade to -9, -12% for the respective speeds of 90 and 105 m.min-1. The group mean net VO2 minimums at -9 and -12%, however, were not significantly different (P > 0.05) from the group mean values at -6 and -15% at 90 m.min-1, or between -9 and -15% grades at 105 m.min-1, Group mean net VO2 values at 0, -3, and -18% were significantly different (P < 0.05) from net VO2 values for the other grades at 90 m.min-1 walking. At 105 m.min-1, mean net VO2 values at 0, -3, -6, and -18% were significantly different (P < 0.05) from net VO2 values at the other grades.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423751 TI - Determinants of VO2peak in patients with end-stage renal disease: on and off dialysis. AB - Peak oxygen uptake (VO2peak) of patients on maintenance hemodialysis is very low. Exercise training performed during, or "on" dialysis, and at other times, "off" dialysis, both improve VO2peak. Equivalence of these approaches has not been shown, nor have the physiologic factors limiting VO2peak been clarified in these patients. We hypothesized that hemodialysis does not acutely alter the cardiovascular response to peak exercise, and further that anemia and a low peak heart rate limit VO2peak. Ten patients with end-stage renal disease (ESRD) performed cycle ergometry to peak exercise. Peak oxygen uptake measured immediately prior to dialysis was compared with predicted values, and with measurements obtained during the second 30 min of dialysis. The determinants of VO2peak were compared with previously reported norms. "Off" dialysis, VO2peak for nine of 10 patients was below the 95% confidence limit for normals. Their cardiovascular response to peak exercise was unchanged by 30-60 min of hemodialysis. Stroke volume was similar to that of normals; however, peak heart rate was 77% of predicted, and hematocrit was 27% (at rest). Peak exercise mixed venous oxygen content was 3.6 ml O2 ml.dl-1 blood, similar to values reported for athletes. These findings show that up to 1 h of dialysis minimally effects VO2peak, and imply that low oxygen delivery limited VO2peak in these patients. PMID- 8423752 TI - Osteoid osteoma of the os calcis in a teenage athlete. AB - Osteoid osteomas are small, benign bone tumors most commonly located in the proximal femur or tibia. The classic presentation is localized pain increasing in severity at night, and relieved by the use of anti-inflammatory medications. In a young athlete complaining of foot pain, many conditions should be included in the differential diagnosis. A case of osteoid osteoma of the os calcis in a teenage basketball player is presented. PMID- 8423753 TI - Tilt table testing in the evaluation and management of athletes with recurrent exercise-induced syncope. AB - Recurrent idiopathic exercise-related syncope in the young athlete is often a challenging and frustrating condition. Vasovagally mediated hypotension and bradycardia is believed to be a common, but difficult to prove, cause of this form of syncope. This study evaluated the usefulness of head-upright tilt table testing in the evaluation and management of young athletes with recurrent idiopathic exercise-related syncope. Twenty-four trained young athletes (12 male, 12 female mean age 18 +/- 3.4 yr) with recurrent unexplained exercise-related syncope were evaluated by use of an upright tilt table test for 30 min, with or without an infusion of isoproterenol (1-3 micrograms.min-1 given intravenously) in an effort to provoke bradycardia, hypotension, or both. Ten control patients with no history of syncope were also studied. Syncope occurred in 10 patients (41%) during the baseline tilt and in nine patients (37%) during the isoproterenol infusion (total positives 79%). Seventeen patients who had positive test results eventually became tilt table negative with pharmacotherapy, and over a mean follow-up period of 23 +/- 7 months, no further syncopal episodes have occurred. Two patients refused pharmacotherapy and have continued to experience syncope. We conclude that head-upright tilt table testing combined with isoproterenol infusion is useful in the diagnosis of vasovagal syncope in young athletes with recurrent exercise related syncope, and in the evaluation of prophylactic pharmacotherapy. PMID- 8423754 TI - Discrimination between cross-country and downhill skiers by pulmonary and local 31PNMR evaluations. AB - The purpose of the present study was to correlate data on calf muscle metabolism using 31P nuclear magnetic resonance spectroscopy with measurements of whole body maximal oxygen consumption and maximal power output, and to determine whether the combination of these data could be used to predict athletic ability. Experiments were performed in a 2.35 Tesla, 35 cm diameter electromagnet on the leg muscle of sedentary human subjects (N = 6) and groups of athletes trained for endurance (cross-country skiers, N = 7) or strength performance (downhill skiers, N = 5). The exercise protocol consisted of successive plantar flexions performed at graded fractions of the maximal voluntary contraction (MVC). The results obtained from NMR investigation (changes in content of inorganic phosphate: Pi, phosphocreatine: PC and muscle ATP, and intracellular pH) were then compared with those of maximal O2 consumption (VO2max) and maximal power (MP). When the data on athletes were compared with those obtained on sedentary subjects, the curves illustrating the relationship between the imposed load and the Pi/PC ratio were significantly shifted toward high output power for a given Pi/PC value. It also appeared from this study that specific training in force development (downhill skiing) induced a slighter decrease in PC level than for endurance (cross-country skiers) despite improvement in physical performance. A slight but significant intracellular acidification was observed in the muscles of sedentary subjects and downhill skiers for contraction at, respectively, 50% and 80% of MVC, but not in the skeletal muscles of cross-country skiers.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423755 TI - Heart rate transient at the onset of active and passive dynamic exercise. AB - At the onset of dynamic exercise there is an almost instantaneous heart rate (HR) acceleration caused by neural activation of central motor areas (central command) and stimulation of mechanoreceptors located in the moving limbs. Aiming to identify the independent contribution of the peripheral mechanism to the initial HR response to exercise, 29 subjects performed two 4-s bouts of unloaded cycling (active (AEx) and passive (PEx)) on an adapted commercial tandem bicycle. PEx was accomplished by having a staff member pedal while sitting on the rear seat. The HR was continuously measured from electrocardiographic tracings. Records of electromyography (EMG) were obtained in a small sample of subjects during the exercise tasks. The number of pedal rotations was very similar (mean +/- SE) (AEx = 7.4 +/- 0.3, PEx = 7.5 +/- 0.2, P = 0.455), determining significant HR changes (P < 0.001) that were similar in the two types of exercise (AEx from 92 to 125 bpm: 35.9% increase; PEx from 87 to 111 bpm: 27.6% increase; P = 0.185). Contrasting to AEx, no muscle contraction was observed by EMG during PEx, suggesting that central command was absent. We concluded that independent activation of mechanoreceptors can promote HR acceleration at the onset of dynamic exercise. PMID- 8423756 TI - Effects of electrolytes in carbohydrate beverages on gastric emptying and secretion. AB - Two experiments were done at rest to examine gastric residue and secretion volume and electrolyte composition after ingestion of beverages of varying composition. In the first experiment the effects of two different sport drinks, one isotonic (7% carbohydrate, primarily sucrose) (I) and one hypertonic (18% carbohydrate, primarily maltodextrin) (H), and a control beverage (0.08 g.l-1 aspartame in water) (C) on titratable acid, pH, osmolality, gastric emptying and secretion volume, and Na+, K+, and Cl- content were measured. In a second experiment five solutions were tested all containing 150 g.l-1 maltodextrin, with 28 meq.l-1 Na+ (low Na), 140 meq.l-1 Na+(high Na), 28 meq.l-1 K+(K), or 140 meq.l-1 Na+ and 28 meq.l-1 K+(high NaK). Beverages H and C, and distilled water (W) were also tested. Samples were taken via a nasogastric tube. A dye dilution technique for serial sampling was employed to determine beverage and secretion volumes. After receiving a bolus of 8 ml.kg-1 body weight, samples of gastric residue were taken at 0, 10, 20, 30, 40, 60, and 80 min. Gastric secretion of Na+, K+, and Cl- was fairly constant despite large differences in beverage composition. Changes in gastric residue pH, titratable acid, osmolality, and electrolyte composition reflected the increasing proportion of the residue that was from gastric secretions. The effects of varying concentrations of Na+ and K+ (in a 150 g.l-1 maltodextrin solution) on gastric emptying were not significant. The high carbohydrate concentration and/or the large volume ingested may have overridden any effect of sodium or potassium. No differences were observed between W and C. Secretion was decreased in these two solutions versus all the others. Although nonsignificant, there was a trend for greater secretion in H versus the other carbohydrate containing solutions in experiment 2. This may be a result of the higher pH maintained after ingestion of this beverage. PMID- 8423757 TI - Exercise training alters feed efficiency and body composition in iron deficient rats. AB - Studies of body composition and feed efficiency were conducted on 80 male Sprague Dawley rats to determine how exercise training alters growth and development in iron deficient animals. Animals were assigned to iron deficient (ID) or control (CN) diets (AIN-76, w/o cellulose), and sedentary (SD) or exercised groups [EX, treadmill running, 4 d.wk-1, 90 min.d-1, 65% maximal oxygen consumption (VO2max), maintained for 6 or 12 wk. The ID diet caused a sustained moderate iron deficiency (Hb 7.2 +/- 0.2 g.dl-1). Iron deficient animals failed to increase maximal oxygen consumption despite the rigorous training program, through training resulted in 25% or 35% higher VO2max in 6- or 12-wk CN rats. At 6 wk, IDEX animals had significantly (33%) lower growth rates than did IDSD animals, which in turn were 22% less than CNs. Overall, exercise did not alter relative amounts of protein in the carcass (% total mass); however, a significant interaction between diet and treatment duration was evident in IDEX animals at 12 wk, who had lower % protein than CNEX-12 or IDSD-6 rats. Training decreased fat 11% in CN at 6 wk but not 12 wk, and 20% in IDs at 12 wk but not 6 wk. Feed efficiency and energy intake were 28% and 12% lower in IDEX animals than CNEX at 6 wk. Training increased caloric intake in CNEX animals but not IDEX animals at 6 and 12 wk. Thus, exercise training exacerbates the poor growth associated with ID through alterations in both food intake and feed efficiency in early phases of training, but adaptation is apparent. PMID- 8423758 TI - Measurement of physical activity to assess health effects in free-living populations. AB - For physical activity surveys that would identify relations to chronic diseases, quality of life, and longevity, the method of choice remains the questionnaire, especially if it can be standardized and administered in uniform fashion to large populations. A sample questionnaire derived largely from epidemiological experience with the Harvard Alumni Health Study is presented that requests anthropometric estimates; physician-diagnosed diseases by year of onset; contemporary physical activities including walking, stair-climbing, and recreational pursuits; food-frequency data that estimate nutrient values and caloric intakes; and social habits affecting health. The questionnaire presents opportunities for cross-sectional, retrospective, and prospective studies. Personal characteristics, physical activities, and other elements of lifestyle may be used as predictor or outcome variables in testing specific hypotheses. Representative surveys are described that have validated and used questionnaires of various complexities, some complemented by measures of physiological fitness. The epidemiological survey questionnaire, when properly designed and administered, can measure effectively energy intake, energy retention, energy expenditure, physiological fitness, quality of life, and health maintenance. PMID- 8423759 TI - A simultaneous evaluation of 10 commonly used physical activity questionnaires. AB - Ten commonly used physical activity questionnaires were evaluated for reliability and validity in 78 men and women aged 20-59, with varying physical activity habits. One month reliability was found to be high for all questionnaires except those pertaining only to the last week or month. Longer term test-retest reliability tended to be lower. Validity was studied in relation to treadmill exercise performance, vital capacity, body fatness, the average of 14 4-wk physical activity histories and the average of 14 2-d accelerometer readings. No questionnaire measure was correlated with the accelerometer reading, and correlations with vital capacity were generally low. Only the Minnesota Leisure Time Physical Activity Questionnaire household chores measure was correlated with habitual performance of household chores. Most questionnaires, even very simple ones, were related to performance of heavy intensity physical activity and treadmill performance; these same questionnaires tended to be related to percent body fat. Fewer questionnaires related to performance of light or moderate activity. Occupational activity was unrelated to any of the validation measures. It is concluded that there are multiple, nonoverlapping dimensions of physical activity, reflected in multiple nonoverlapping validation realms. More important than the length or attention to detail of a questionnaire seems to be the logic of its questions. Important areas of physical activity that should be addressed in future questionnaires include sleep, light, moderate and heavy intensity leisure activities, household chores, and occupational activity. Recent versus habitual activity should also be considered. PMID- 8423760 TI - Effect of ibuprofen use on muscle soreness, damage, and performance: a preliminary investigation. AB - Twenty subjects were randomly assigned to: 1) prophylactic ibuprofen (N = 5) [400 mg TID initiated 4 h before collection of baseline data and strenuous eccentric exercise bout], 2) therapeutic ibuprofen (N = 5) [400 mg TID initiated 24 h after baseline], 3) placebo (N = 5), or 4) control (N = 5). Muscle soreness perception, plasma creatine kinase, knee extensor torque, and EMG of the quadriceps were evaluated at baseline, 24, and 48 h. The prophylactic ibuprofen group had between 40 and 50% less muscle soreness perception and significantly less decline in isometric, concentric, and eccentric torque at 24 h compared with the other three groups (P < 0.05). At 48 h both prophylactic and therapeutic ibuprofen had significantly less muscle soreness perception and decline in torque than the placebo and control groups (P < 0.05). There was no difference between the amount of muscle damage between the four groups at 24 and 48 h. Vastus medialis and lateralis EMG magnitude decreased across time. Vastus lateralis EMG magnitude had significantly less decline from baseline for prophylactic ibuprofen compared with the other three treatments at 24 h, while both prophylactic and therapeutic ibuprofen had significantly less decline at 48 h. These data indicate that a prophylactic dosage of ibuprofen does not prevent CK release from muscle, but does decrease muscle soreness perception and may assist in restoring muscle function. PMID- 8423761 TI - Validity and reliability of self-reported physical activity status: the Lipid Research Clinics questionnaire. AB - We assessed the validity and reliability of the Lipid Research Clinics (LRC) physical activity (PA) questionnaire against measures of PA and physical fitness, and compared a new scoring system (four-point) with the method used in the LRC studies (two-point) in 28 men and 50 women. The two-point method classified participants as inactive or active based one question about regular, strenuous exercise habits. The four-point method classified participants as very low, low, moderate, and high active based on two questions; regular, strenuous exercise and self-rating of PA relative to peers. After adjusting for age and gender, the four point method explained 17-29% of the variation in VO2max, percent body fat, and heavy-and light-intensity kcal.d-1 measured by the 4 wk history questionnaire (FWH). No association was found between the LRC ratings and Caltrac kcal and activity units and FWH total, moderate-intensity, and household PA kcal.d-1. Test retest reliability was high (two-point, r = 0.85; four-point, r = 0.88). The LRC PA questionnaire is a relatively valid and reliable indicator of heavy PA in adults. The four-point method increases the ability to show graded associations between measures of cardiorespiratory fitness, body fatness, and leisure time PA. PMID- 8423762 TI - Seven-day recall and other physical activity self-reports in children and adolescents. AB - There is need to develop low cost, practical, and accurate measures of physical activity in children and adolescents, and self-report is a promising methodology for children that is applicable for large studies. The purpose of the present study was to assess the reliability and validity of several self-reports of physical activity. Subjects were 36 fifth-, 36 eighth-, and 30 eleventh-grade male and female students. The test-retest reliabilities were r = 0.77 for the 7-d recall interview, r = 0.81 for the Godin-Shephard self-administered survey, and r = 0.93 for a simple activity rating. For the former two measures, reliability improved with age but was significant at all ages, and for the last measure there were no age effects. Memory skills and obesity status were not related to the reliability of recall, but males were more reliable reporters than females. Validity of the 7-d recall was determined by comparing heart rate monitoring records with recalls of very hard activities on the same day. A correlation of 0.53 (P < 0.001) for the total group supported the validity of the reports. Validity improved with age, but validity coefficients were significant in all age groups. These data indicate that physical activity recalls of children as young as the fifth grade are of adequate reliability and validity to use in research on physical activity in children. PMID- 8423763 TI - Zinc modulation of drug binding, cocaine affinity states, and dopamine uptake on the dopamine uptake complex. AB - Zinc plays an important role in synaptic function in the brain. Zinc is present in synaptic vesicles, is released with neuronal activity, and provides modulation of different neurotransmitter systems. Zinc altered characteristics of the dopamine uptake complex in rat caudate-putamen. The density of the dopamine uptake complex labeled with [3H]GBR 12935 (0.25 nM) was increased by up to 215% in the presence of zinc (10 microM), compared with control. Augmentation of binding occurred with concentrations of zinc from 0.5 to 100 microM. This effect on the dopamine uptake complex was due to a 65% decrease in the equilibrium dissociation constant for [3H]GBR 12935 without a significant change in the total number of binding sites. The presence of exogenous zinc (50 microM) increased the percentage of high affinity sites for cocaine from 73 +/- 4% to 90 +/- 1% and decreased the high affinity constant from 2250 +/- 880 nM to 890 +/- 150 nM. Similar effects of zinc occurred with the cocaine congener 2 beta-carbomethoxy-3 beta-(fluorophenyl)tropane. Zinc inhibited dopamine uptake into synaptosomes at concentrations from 0.5 to 100 microM. Zinc (0.5 and 1 microM) augmented cocaine inhibition of uptake by 58-79%. Modulation of drug binding to the dopamine uptake complex is another role for zinc in the central nervous system. Sulfhydryl oxidation of the dopamine uptake complex by certain metal cations (Cu2+, Hg2+, and Cd2+) or N-methylmaleimide reduced binding of [3H]GBR 12935 to the dopamine uptake complex. The sulfhydryl oxidation by N-methylmaleimide was blocked by coincubation with dithiothreitol and dopamine uptake blockers. These data support the existence of a sulfhydryl group involved in binding of uptake blockers to the dopamine uptake complex. PMID- 8423764 TI - 4-Methylpyrazole inhibits fatty acyl coenzyme synthetase and diminishes catalase dependent alcohol metabolism: has the contribution of alcohol dehydrogenase to alcohol metabolism been previously overestimated? AB - Alcohol dehydrogenase (ADH)-deficient deer mice were used as an animal model to investigate the effect of 4-methylpyrazole on alcohol metabolism. After intraperitoneal dosing of these mutant mice with 4-methylpyrazole, rates of ethanol and methanol metabolism in vivo were decreased significantly, by 41% and 35%, respectively. In perfused liver, rates of ethanol metabolism were also decreased up to 61% by 100 microM 4-methylpyrazole. Further, when livers were perfused with methanol, a selective substrate for catalase, rates of methanol metabolism were decreased by 64% by 4-methylpyrazole. It was further determined that 4-methylpyrazole administration caused negligible changes in total hepatic catalase activity and in rates of oxidation of ethanol by isolated microsomes; rather, it acts on catalase-dependent alcohol metabolism by limiting the supply of H2O2. In this study, 4-methylpyrazole inhibited fatty acyl CoA synthetase competitively in liver homogenates. Fatty acyl CoA synthetase is a key enzyme involved in the supply of substrate for peroxisomal oxidation of alcohols via catalase-H2O2. When palmitate was studied, rates of formaldehyde production from methanol were reduced competitively by 4-methylpyrazole; however, when the product palmitoyl CoA was used, the addition of 4-methylpyrazole did not alter activity. 4-Methylpyrazole also inhibited fatty acyl CoA synthetase activity measured directly from CoA disappearance. These data indicate that fatty acyl CoA synthetase is inhibited by 4-methylpyrazole, thus reducing the availability of H2O2 for catalase-dependent alcohol metabolism. Inhibition of methanol metabolism in deer mice expressing ADH indicates that this phenomenon also occurs in species with ADH. Taken together, these data support the hypothesis that the contribution of ADH to alcohol metabolism may have been previously overestimated. PMID- 8423765 TI - Identification and characterization of the cytochrome P450 enzymes involved in N dealkylation of propafenone: molecular base for interaction potential and variable disposition of active metabolites. AB - The activity of metabolizing enzymes determines plasma concentrations and hence effects of drugs. Identification of these enzymes may allow the prediction of both the interaction potential of drugs and the variability deriving from certain pathways. The antiarrhythmic propafenone is extensively biotransformed to the active metabolites 5-hydroxypropafenone and N-desalkylpropafenone. Whereas 5 hydroxylation is catalyzed by CYP2D6, the enzyme involved in N-dealkylation has not been identified. We, therefore, characterized the enzyme involved in the formation of N-desalkylpropafenone by using both in vitro [human liver microsomes, specific antibodies or inhibitors, and stably expressed cytochrome P450 (P450) enzymes] and in vivo (formation of N-desalkylpropafenone in patients under conditions of chronic therapy) approaches. Formation of N desalkylpropafenone can be described by Michaelis-Menten kinetics. A strong correlation was observed between maximum rate of formation (Vmax) of N desalkylpropafenone and the amount of CYP1A2 (r = 0.83, p < 0.001) and CYP3A (r = 0.54, p < 0.05) in the microsomal fraction of 20 human livers. In vitro intrinsic clearances (derived from Vmax/Km) indicated a wide interindividual variability in seven human livers (from 0.01 to 0.1 ml/hr/mg of protein). Antibodies directed against CYP3A and CYP1A2 inhibited formation of N-desalkylpropafenone by 54 +/- 10% and 24 +/- 16%, respectively. The CYP2D6-mediated formation of 5 hydroxypropafenone was unaffected by these antibodies. Verapamil (substrate of CYP3A4 and CYP1A2) and midazolam (substrate of CYP3A4) were competitive inhibitors of N-desalkylpropafenone formation (Ki = 70 microM and 25 microM for verapamil and midazolam, respectively). Coding sequences for CYP1A2 and CYP3A4 were inserted in a yeast expression vector and introduced into Saccharomyces cerevisiae strain W(R). Both CYP1A2 and CYP3A4 catalyzed N-dealkylation of propafenone, with specific activities of 0.32 pmol/min/pmol of P450 and 0.16 pmol/min/pmol of P450, respectively. Our data indicate that N-dealkylation of propafenone is mediated via CYP3A4 and CYP1A2. From experiments on the molecular level interactions of propafenone with other drugs that are metabolized by CYP3A4 and CYP1A2 can be predicted. Such interactions have been reported for cyclosporin, rifampicin, warfarin, and theophylline. Moreover, in vitro intrinsic clearances showed a wide interindividual variability. Therefore, variable plasma concentrations of the active metabolite N-desalkylpropafenone are expected in vivo. We tested this hypothesis in 14 patients (dose of 150 mg of propafenone three times per day) during chronic oral therapy and observed steady state plasma concentrations of N-desalkylpropafenone ranging from 4 to 293 ng/ml.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8423766 TI - Solubilization of the cannabinoid receptor from rat brain and its functional interaction with guanine nucleotide-binding proteins. AB - The present investigation was undertaken to characterize cannabinoid receptor binding in the absence of the membrane environment, inasmuch as cannabinoid drugs have been noted to influence the behavior of integral membrane proteins. The zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was able to solubilize the cannabinoid receptor from rat brain membranes, with the greatest yield and specific activity being obtained at a detergent/protein ratio of 0.5:1. [3H]CP-55940 bound to a single class of binding sites in the CHAPS extract, which exhibited a Kd of 0.94 nM as determined by nonlinear regression analysis of equilibrium binding data. The order of potency for cannabinoid agonists in heterologous equilibrium binding studies was CP-55244 > or = desacetyllevonantradol > delta 9-tetrahydrocannabinol > cannabinol >> cannabidiol, consistent with the relative affinities for these agonists in brain membrane preparations. CP-55243, the biologically inactive enantiomer of CP 55244, competed for binding of [3H]CP-55940 by < 50% at 1 microM, similar to its poor affinity for the receptor in membranes. The CHAPS-solubilized cannabinoid receptor exhibited functional interactions with guanine nucleotide-binding proteins (G proteins). GTP and nonhydrolyzable analogs decreased [3H]CP-55940 binding by 75%. The concentration-effect curves for guanine nucleotides exhibited a potency order similar to that observed for other G protein-linked receptors. Kinetic analyses indicated that GTP analogs increased the rate of agonist dissociation, decreasing the t1/2 from 60 min at 0-4 degrees to a multiphasic dissociation that exhibited a component having a t1/2 of < 1 min. The cannabinoid agonist desacetyllevonantradol was able to reduce pertussis toxin-catalyzed ADP ribosylation of G proteins by 50%, demonstrating a receptor effect on G protein functions. These studies demonstrate that the membrane environment is not necessary for agonist binding to the cannabinoid receptor. Furthermore, the cannabinoid receptor maintains its functional interactions with pertussis toxin sensitive G proteins in detergent solution. PMID- 8423767 TI - High-dose chemotherapy-induced platelet defect: inhibition of platelet signal transduction pathways. AB - Patients receiving high-dose chemotherapy and autologous bone marrow transplantation acquire a platelet secretion defect. The role of chemotherapeutic agents and their metabolites in mediating this platelet defect was investigated. 1,3-Bis(2-chloroethyl)-1-nitrosourea (BCNU), but not cyclophosphamide or cis platinum, was found to inhibit platelet aggregation in vitro in response to activation by either ADP, thrombin, or collagen. Inhibition by BCNU was dose dependent and required preincubation of platelets with BCNU. After a 60-min preincubation, 30 microM BCNU produced 50% inhibition of platelets in platelet rich plasma. The cyclophosphamide metabolites acrolein and 4 hydroperoxycyclophosphamide also inhibited platelet aggregation in a dose dependent manner, with a requirement for preincubation. Platelet inhibition occurred at clinically relevant concentrations of BCNU and metabolites of cyclophosphamide. The effects of acrolein were totally prevented by coincubation with the sulfhydryl-protecting agents N-acetylcysteine and 2 mercaptoethanesulfonic acid, whereas the effects of BCNU were incompletely prevented. The mechanism of platelet inhibition was investigated next by examining protein phosphorylation in response to platelet agonists. Acrolein inhibited thrombin- and phorbol ester-induced phosphorylation of a 40-kDa polypeptide and other substrates, indicating a cellular defect in protein kinase C signaling. BCNU did not interfere with protein phosphorylation, indicating preservation of initial signaling pathways. Thus, chemotherapeutic agents and their metabolites inhibit platelet function by inhibiting distinct components of the intracellular activation pathways. PMID- 8423768 TI - Cellular pharmacology of phosphorothioate homooligodeoxynucleotides in human cells. AB - The uptake and distribution of phosphorothioate oligodeoxynucleotides by human cells were studied using 35S-labeled 28-mer phosphorothioate oligodeoxycytidine [S-(dC)28]. Accumulation of intracellular S-(dC)28 was found to be higher in the carcinoma cells (grown in monolayers) than in the leukemia cells (grown in suspension culture). A hepatoma cell line transfected with hepatitis B virus, 2215, was chosen for further studies. The uptake of S-(dC)28 was partially dependent on temperature and energy. The intracellular concentration was significantly higher than that in the medium and the amount accumulated was dependent on the extracellular concentration. It appears that the uptake of S (dC)28 involves mechanisms of both fluid-phase pinocytosis and adsorptive endocytosis. Neither oligonucleotides nor 5'-phosphorylated nucleotides inhibited S-(dC)28 uptake. Unlike horseradish peroxidase, which was primarily associated with endosomes once it was taken into the cell, S-(dC)28 was found to be present in both nuclear and cytoplasmic fractions. Efflux of S-(dC)28 from the cell was multiphasic; a trapping mechanism that could be due to a potent interaction of S (dC)28 with cellular proteins was implicated. This trapping mechanism could be responsible for the lack of biological activity such as cytotoxicity and antisense activity of phosphorothioate oligodeoxynucleotides in some human cells. PMID- 8423769 TI - Estradiol induction of rhodamine 123 efflux and the multidrug resistance pump in rat pituitary tumor cells. AB - Rhodamine 123 is a fluorescent dye that localizes in mitochondria, is a substrate for the multidrug resistance pump, and is retained for long periods of time by carcinoma cells. 17 beta-Estradiol causes GH4C1 cells (rat pituitary tumor cells) to lose rhodamine 123 fluorescence faster than untreated cells. We found that estradiol induces accumulation of the mRNA for the multidrug resistance pump 3-5 fold, with maximum induction occurring within 1 day at 10(-9) M estradiol. Immunoblot analysis demonstrated that estradiol induces a protein of 150 kDa that reacts with an antibody to P-glycoprotein, the multidrug resistance pump. The reduced retention of rhodamine 123 caused by estradiol is prevented by verapamil and cyclosporin, inhibitors of the pump. A clone resistant to the effects of estradiol on rhodamine 123 has greatly reduced levels of mRNA for the pump. The effect of estradiol is more marked on rhodamine 123 retention than it is on that of rhodamine 110 or tetramethylrhodamine methyl ester. We conclude that estradiol enhances rhodamine 123 efflux by inducing the multidrug resistance gene. The specificity for rhodamine 123, compared with other analogs, may be caused by differences in accessibility to the pump. PMID- 8423770 TI - Sequence analysis and expression of the cDNA for the phenol-sulfating form of human liver phenol sulfotransferase. AB - A cDNA encoding the human liver phenol-sulfating form of phenol sulfotransferase (P-PST) has been isolated and characterized from a lambda Uni-Zap XR human liver cDNA library. P-PST is the major form of phenol sulfotransferase involved in drug and xenobiotic metabolism in human liver. P-PST is also responsible for the sulfation and activation of minoxidil to its therapeutically active sulfate ester. The full length cDNA, P-PST-1, is 1206 base pairs in length and encodes a 295-amino acid protein with a molecular mass of 34,097 Da. The translation sequence of P-PST-1 is 96% similar to the amino acid sequences of five peptides derived from the purified protein. In vitro transcription and translation of P PST-1 generated a protein that comigrates with immunoreactive P-PST from human liver. Significant increases in sulfotransferase activity toward two P-PST specific substrates, minoxidil and 4-nitrophenol, were detected in cytosol prepared from COS-7 cells transfected with P-PST-1 in the expression vector p-SV SPORT-1. Northern blot analysis of human liver RNA detected a transcript of approximately 1300 nucleotides in length. Characterization of P-PST at the molecular level provides insight into the structure and heterogeneity of this major class of drug-metabolizing enzymes. PMID- 8423771 TI - Hallucinogenic receptor models: interaction of imidazolium chloride with amphetamine analogs. AB - The interactions of several hallucinogenic phenethylamines with the biologically relevant electron acceptor imidazolium chloride have been investigated by monitoring phenethylamine 1H NMR chemical shift changes upon complex formation. Methoxy- and methylenedioxy-substituted phenethylamines interact with imidazolium chloride to form weak charge-transfer complexes that have a relatively narrow range of association constants. The results indicate the formation of a ternary complex in which a chloride ion is hydrogen bonded to a N+H side chain proton of the drug and a NH imidazolium proton. The imidazolium ring is positioned above a nonsubstituted ortho-position of the aromatic ring. Spectral assignment of the two diastereotopic benzylic protons allowed the geometry of the complex to be defined in more detail. Complexation occurs preferentially on that side of the phenethylamine molecule that allows the side chain alpha-methyl group to face away from the chloride ion and the imidazolium ring. The drug molecule exists in the complex in the preferred trans conformation. The biological relevance of this model is discussed. PMID- 8423772 TI - Repeated administration of SR 46349B, a selective 5-hydroxytryptamine2 antagonist, up-regulates 5-hydroxytryptamine2 receptors in mouse brain. AB - Adaptive changes in 5-hydroxytryptamine (5-HT)2 receptors were investigated in mice after repeated administration of SR 46349B, a potent, selective, and competitive 5-HT2 receptor antagonist (Kl = 0.72 +/- 0.05 nM). Repeated administration (twice per day for 3 days and once on the morning of the fourth day) of SR 46349B (5 or 10 mg/kg, orally) caused 24 hr later a marked increase in 5-HT2 receptor number (+41% and +75%, respectively), measured ex vivo in brain cortical membranes with [3H] ketanserin, without affecting its affinity constant. Further, administration of the 5-HT2 agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl) 2-aminopropane produced, in SR 46349B (10 mg/kg, orally)-treated mice, a significant stimulation of the 5-HT2 receptor-linked phosphoinositide turnover in vivo in the brain. In addition, subacute administration of SR 46349B (5 or 10 mg/kg, orally) caused a significant increase of the head-twitch response to L-5 hydroxytryptophan. This enhanced response was blocked by an acute administration of ritanserin (1 mg/kg). These results show that repeated administration of SR 46349B produced a parallel enhancement in 5-HT2 receptor number, in 5-HT2 receptor-linked signal transduction, and in 5-HT2 receptor-mediated behavioral responses in mice. These findings suggest for the first time that an up regulation of 5-HT2 receptors can occur after repeated treatment with a selective 5-HT2 antagonist. PMID- 8423773 TI - Defective replication of psoralen adducts detected at the gene-specific level in xeroderma pigmentosum variant cells. AB - Replication of damaged DNA is suspected to play an important role in cell cycle, genetic stability, and survival pathways. Using psoralen photoaddition as prototype DNA damage and the renaturing agarose gel electrophoresis technique to measure DNA cross-linking in individual genes, Vos and Hanawalt previously observed efficient bypass replication of psoralen monoadducts in human genes (J. M. H. Vos and P. C. Hanawalt, Cell 50:789-799, 1987). To understand the mechanism of bypass replication in human cells, mutants affected in such a process would be useful. We now report that cells from individuals suffering from the hereditary recessive syndrome xeroderma pigmentosum variant (XPV) are hypersensitive to killing induced by photoactivated psoralen. In addition, analysis of psoralen mediated DNA cross-linking in the rRNA genes indicated that although repair of psoralen adducts was similar to that of normal individuals, XPV cells were markedly deficient in the ability to bypass psoralen adducts during replication; in comparison with normal cells, approximately half as many monoadducts were bypassed during replication in XPV cells. Furthermore, in contrast to normal cells, replication of interstrand cross-links was not detected in XPV. This is the first demonstration of a deficiency in bypass replication detected at the gene-specific level in vivo. A model involving a strand-specific defect in recombinational bypass in XPV is proposed. PMID- 8423774 TI - FAR1 is required for posttranscriptional regulation of CLN2 gene expression in response to mating pheromone. AB - Yeast cells arrest during the G1 interval of the cell cycle in response to peptide mating pheromones. The FAR1 gene is required for cell cycle arrest but not for a number of other aspects of the pheromone response. Genetic evidence suggests that FAR1 is required specifically for inactivation of the G1 cyclin CLN2. From these observations, the FAR1 gene has been proposed to encode an element of the interface between the mating pheromone signal transduction pathway and the cell cycle regulatory apparatus. We show here that FAR1 is necessary for the decrease in CLN1 and CLN2 transcript accumulation observed in response to mating pheromone but is unnecessary for regulation of the same transcripts during vegetative growth. However, the defect in regulation of CLN1 expression is dependent upon CLN2. We show that pheromone regulates the abundance of Cln2 at a posttranscriptional level and that FAR1 is required for that regulation. From these observations, we suggest that FAR1 function is limited to posttranscriptional regulation of CLN2 expression by mating pheromone. The failure of mating pheromone to repress CLN2 transcript levels in far1 mutants can be explained by the stimulatory effect of the persistent Cln2 protein on CLN2 transcription via the CLN/CDC28-dependent feedback pathway. PMID- 8423775 TI - DNA repair synthesis during base excision repair in vitro is catalyzed by DNA polymerase epsilon and is influenced by DNA polymerases alpha and delta in Saccharomyces cerevisiae. AB - Base excision repair is an important mechanism for correcting DNA damage produced by many physical and chemical agents. We have examined the effects of the REV3 gene and the DNA polymerase genes POL1, POL2, and POL3 of Saccharomyces cerevisiae on DNA repair synthesis is nuclear extracts. Deletional inactivation of REV3 did not affect repair synthesis in the base excision repair pathway. Repair synthesis in nuclear extracts of pol1, pol2, and pol3 temperature sensitive mutants was normal at permissive temperatures. However, repair synthesis in pol2 nuclear extracts was defective at the restrictive temperature of 37 degrees C and could be complemented by the addition of purified yeast DNA polymerase epsilon. Repair synthesis in pol1 nuclear extracts was proficient at the restrictive temperature unless DNA polymerase alpha was inactivated prior to the initiation of DNA repair. Thermal inactivation of DNA polymerase delta in pol3 nuclear extracts enhanced DNA repair synthesis approximately 2-fold, an effect which could be specifically reversed by the addition of purified yeast DNA polymerase delta to the extract. These results demonstrate that DNA repair synthesis in the yeast base excision repair pathway is catalyzed by DNA polymerase epsilon but is apparently modulated by the presence of DNA polymerases alpha and delta. PMID- 8423776 TI - Analysis of the SWI4/SWI6 protein complex, which directs G1/S-specific transcription in Saccharomyces cerevisiae. AB - SWI4 and SWI6 play a crucial role in START-specific transcription in Saccharomyces cerevisiae. SWI4 and SWI6 form a specific complex on the SCB (SWI4/6-dependent cell cycle box) sequences which have been found in the promoters of HO and G1 cyclin genes. Overproduction of SWI4 eliminates the SWI6 dependency of HO transcription in vivo and results in a new SWI6-independent, SCB specific complex in vitro, which is heterogeneous and reacts with SWI4 antibodies. The C terminus of SWI4 is not required for SWI6-independent binding of SWI4 to SCB sequences, but it is necessary and sufficient for association with SWI6. Both SWI4 and SWI6 contain two copies of a 33-amino-acid TPLH repeat, which has been implicated in protein-protein interactions in other proteins. These repeats are not required for the SWI4-SWI6 association. Alanine substitutions in both TPLH repeats of SWI6 reduce its activity but do not affect the stability of the protein or its association with SWI4. However, these mutations reduce the ability of the SWI4/6 complex to bind DNA. Deletion of the lucine zipper motif in SWI6 also allows SWI4/6 complex formation, but it eliminates the DNA-binding ability of the SWI4/6 complex. This indicates that the integrity of two different regions of SWI6 is required for DNA binding by the SWI4/6 complex. From these data, we propose that the sequence-specific DNA-binding domain resides in SWI4 but that SWI6 controls the accessibility of this domain in the SWI4/6 complex. PMID- 8423777 TI - Erythroid differentiation of mouse erythroleukemia cells results in reorganization of protein-DNA complexes in the mouse beta maj globin promoter but not its distal enhancer. AB - Dimethyl sulfoxide (DMSO) induction of mouse erythroleukemia (MEL) cells represents a well-defined in vitro system of terminal erythroid differentiation. We have studied the molecular mechanisms of transcriptional activation of the mouse beta maj globin gene during MEL cell differentiation by analyzing nuclear factor-DNA interactions in vivo at the gene's upstream promoter and a distal enhancer, 5'HS-2. Genomic footprinting data indicate that three motifs, CAC, NF E2/AP1, and GATA-1, of the 5'HS-2 enhancer are bound with nuclear factors in MEL cells both prior to and after DMSO induction. No obvious conformational change of these nuclear factor-DNA complexes could be detected upon terminal differentiation of MEL cells. On the other hand, DMSO induction of MEL cells leads to the formation of specific nuclear factor-DNA complexes at several transcriptional regulatory elements of the mouse beta maj globin upstream promoter. Our genomic footprinting data have interesting implications with respect to the molecular mechanisms of transcriptional regulation and chromatin change of the mouse beta maj globin gene during erythroid differentiation. PMID- 8423778 TI - Species-specific signals for the splicing of a short Drosophila intron in vitro. AB - The effects of branchpoint sequence, the pyrimidine stretch, and intron size on the splicing efficiency of the Drosophila white gene second intron were examined in nuclear extracts from Drosophila and human cells. This 74-nucleotide intron is typical of many Drosophila introns in that it lacks a significant pyrimidine stretch and is below the minimum size required for splicing in human nuclear extracts. Alteration of sequences of adjacent to the 3' splice site to create a pyrimidine stretch was necessary for splicing in human, but not Drosophila, extracts. Increasing the size of this intron with insertions between the 5' splice site and the branchpoint greatly reduced the efficiency of splicing of introns longer than 79 nucleotides in Drosophila extracts but had an opposite effect in human extracts, in which introns longer than 78 nucleotides were spliced with much greater efficiency. The white-apricot copia insertion is immediately adjacent to the branchpoint normally used in the splicing of this intron, and a copia long terminal repeat insertion prevents splicing in Drosophila, but not human, extracts. However, a consensus branchpoint does not restore the splicing of introns containing the copia long terminal repeat, and alteration of the wild-type branchpoint sequence alone does not eliminate splicing. These results demonstrate species specificity of splicing signals, particularly pyrimidine stretch and size requirements, and raise the possibility that variant mechanisms not found in mammals may operate in the splicing of small introns in Drosophila and possibly other species. PMID- 8423779 TI - U2 small nuclear RNA 3' end formation is directed by a critical internal structure distinct from the processing site. AB - Mature U2 small nuclear RNA is generated by the removal of 11 to 12 nucleotides from the 3' end of the primary transcript. This pre-U2 RNA processing reaction takes place in the cytoplasm. In this study, the sequences and/or structures of pre-U2 RNA that are important for 3' processing have been examined in an in vitro system. The 7-methylguanosine cap, stem-loops I and II, the lariat branch site recognition sequence, the conserved Sm domain, and several other regions throughout the 5' end of U2 RNA have no apparent role in the 3' processing reaction. In fact, deletion of the entire first 104 nucleotides resulted in mini pre-U2 RNAs which were efficiently processed. Similarly, deletion of the top two thirds of stem-loop III or mutation of nucleotides in the loop of stem-loop IV had little effect on 3' processing. Most surprisingly, the precursor's 11- to 12 nucleotide 3' extension itself was of relatively little importance, since this sequence could be replaced with completely different sequences with only a minor effect on the 3' processing reaction. In contrast, we have defined a critical structure consisting of the bottom of stem III and the stem of stem-loop IV that is essential for 3' processing of pre-U2 RNA. Compensatory mutations which restore base pairing in this region resulted in normal 3' processing. Thus, although the U2 RNA processing activity recognizes the bottom of stem III and stem IV, the sequence of this critical region is much less important than its structure. These results, together with the surprising observation that the reaction is relatively indifferent to the sequence of the 11- to 12-nucleotide 3' extension itself, point to a 3' processing reaction of pre-U2 RNA that has sequence and structure requirements significantly different from those previously identified for pre-mRNA 3' processing. PMID- 8423780 TI - B-myc inhibits neoplastic transformation and transcriptional activation by c-myc. AB - B-myc is a recently described myc gene whose product has not been functionally characterized. The predicted product of B-myc is a 168-amino-acid protein with extensive homology to the c-Myc amino-terminal region, previously shown to contain a transcriptional activation domain. We hypothesized that B-Myc might also function in transcriptional regulation, although its role in regulating gene expression is predicted to be unique, because B-Myc lacks the specific DNA binding motif found in other Myc proteins. To determine whether B-Myc could interact with the transcriptional machinery, we studied the transcriptional activation properties of a chimeric protein containing B-Myc sequences fused to the DNA-binding domain of the yeast transcriptional activator GAL4 (GAL4-B-Myc). We found that GAL4-B-Myc strongly activated expression of a GAL4-regulated reporter gene in mammalian cells. In addition, full-length B-Myc was able to inhibit or squelch reporter gene activation by a GAL4 chimeric protein containing the c-Myc transcriptional activation domain. We also observed that B-Myc dramatically inhibited the neoplastic cotransforming activity of c-Myc and activated Ras in rat embryo cells. Because B-Myc inhibits both neoplastic transformation and transcriptional activation by c-Myc, we suggest that the transforming activity of c-Myc is related to its ability to regulate transcription. Whether B-Myc functions biologically to squelch transcription and/or to regulate transcription through a specific DNA-binding protein remains unestablished. PMID- 8423781 TI - Differential regulation of two distinct families of glucose transporter genes in Trypanosoma brucei. AB - A tandemly arranged multigene family encoding putative hexose transporters in Trypanosoma brucei has been characterized. It is composed of two 80% homologous groups of genes called THT1 (six copies) and THT2 (five copies). When Xenopus oocytes are microinjected with in vitro-transcribed RNA from a THT1 gene, they express a glucose transporter with properties similar to those of the trypanosome bloodstream-form protein(s). This THT1-encoded transport system for glucose differs from the human erythrocyte-type glucose transporter by its moderate sensitivity to cytochalasin B and its capacity to transport D-fructose. These properties suggest that the trypanosomal transporter may be a good target for antitrypanosomal drugs. mRNA analysis revealed that expression of these genes was life cycle stage dependent. Bloodstream forms express 40-fold more THT1 than THT2. In contrast, procyclic trypanosomes express no detectable THT1 but demonstrate glucose-dependent expression of THT2. PMID- 8423782 TI - Transforming growth factor beta and cyclosporin A inhibit the inducible activity of the interleukin-2 gene in T cells through a noncanonical octamer-binding site. AB - Transforming growth factor beta (TGF-beta) has a growth-inhibitory effect on numerous different cell types of the immune system, including T lymphocytes. We show in this study that the inhibitory action of TGF-beta on T lymphocytes is accompanied by a block of interleukin 2 (IL-2) gene expression which is mediated, at least in part, by inhibition of IL-2 promoter/enhancer activity. The functional analysis of cis-regulatory (proto-enhancer) elements of the IL-2 enhancer/promoter region showed that the most TGF-beta-responsive element maps to its so-called upstream promoter site. The proto-enhancer activity of the upstream promoter site element is also inhibited by cyclosporin A. The upstream promoter site DNA harbors two noncanonical, closely linked binding sequences for octamer and AP-1-like factors. Both sites are involved in the establishment of IL-2 enhancer activity. Since the activity of genuine octamer sites but not that of AP 1-binding sites is also impaired by TGF-beta and cyclosporin A in El4 T lymphoma cells, we conclude that both immunosuppressives interfere with the activity but not the DNA binding of octamer factors in T lymphocytes. PMID- 8423783 TI - Biochemical analysis of torso and D-raf during Drosophila embryogenesis: implications for terminal signal transduction. AB - Determination of anterior and posterior terminal structures of Drosophila embryos requires activation of two genes encoding putative protein kinases, torso and D raf. In this study, we demonstrate that Torso has intrinsic tyrosine kinase activity and show that it is transiently tyrosine phosphorylated (activated) at syncytial blastoderm stages. Torso proteins causing a gain-of-function phenotype are constitutively tyrosine phosphorylated, while Torso proteins causing a loss of-function phenotype lack tyrosine kinase activity. The D-raf gene product, which is required for Torso function, is identified as a 90-kDa protein with intrinsic serine/threonine kinase activity. D-Raf is expressed throughout embryogenesis; however, the phosphorylation state of the protein changes during development. In wild-type embryos, D-Raf is hyperphosphorylated at 1 to 2 h after egg laying, and thereafter only the most highly phosphorylated form is detected. Embryos lacking Torso activity, however, show significant reductions in D-Raf protein expression rather than major alterations in the protein's phosphorylation state. This report provides the first biochemical analysis of the terminal signal transduction pathway in Drosophila embryos. PMID- 8423784 TI - A bipartite DNA-binding domain in yeast Reb1p. AB - The REB1 gene encodes a DNA-binding protein (Reb1p) that is essential for growth of the yeast Saccharomyces cerevisiae. Reb1p binds to sites within transcriptional control regions of genes transcribed by either RNA polymerase I or RNA polymerase II. The sequence of REB1 predicts a protein of 809 amino acids. To define the DNA-binding domain of Reb1p, a series of 5' and 3' deletions within the coding region was constructed in a bacterial expression vector. Analysis of the truncated Reb1p proteins revealed that nearly 400 amino acids of the C terminal portion of the protein are required for maximal DNA-binding activity. To further define the important structural features of Reb1p, the REB1 homolog from a related yeast, Kluyveromyces lactis, was cloned by genetic complementation. The K. lactis REB1 gene supports active growth of an S. cerevisiae strain whose REB1 gene has been deleted. The Reb1p proteins of the two organisms generate almost identical footprints on DNA, yet the K. lactis REB1 gene encodes a polypeptide of only 595 amino acids. Comparison of the two Reb1p sequences revealed that within the region necessary for the binding of Reb1p to DNA were two long regions of nearly perfect identity, separated in the S. cerevisiae Reb1p by nearly 150 amino acids but in the K. lactis Reb1p by only 40 amino acids. The first includes a 105 amino-acid region related to the DNA-binding domain of the myb oncoprotein; the second bears a faint resemblance to myb. The hypothesis that the DNA-binding domain of Reb1p is formed from these two conserved regions was confirmed by deletion of as many as 90 amino acids between them, with little effect on the DNA binding ability of the resultant protein. We suggest that the DNA-binding domain of Reb1p is made up of two myb-like regions that, unlike myb itself, are separated by as many as 150 amino acids. Since Reb1p protects only 15 to 20 nucleotides in a chemical or enzymatic footprint assay, the protein must fold such that the two components of the binding site are adjacent. PMID- 8423785 TI - The human beta fibrinogen promoter contains a hepatocyte nuclear factor 1 dependent interleukin-6-responsive element. AB - Acute-phase reactants are liver proteins whose synthesis is positively or negatively regulated during inflammation. The main mediators of this phenomenon are glucocorticoids and interleukin-6 (IL-6), a pleiotropic cytokine that also controls hematopoiesis. Functional analysis of several acute-phase reactant promoter regions has identified two major DNA motifs used by IL-6-regulated genes. The first one corresponds to a CTGG(G/A)AA sequence, and the other is a binding site for members of the C/EBP family of nuclear proteins. We have previously shown that the human beta fibrinogen (beta Fg) promoter contains an IL 6-responsive region, located between bp -150 and -67 (P. Huber, M. Laurent, and J. Dalmon, J. Biol. Chem. 265:5695-5701, 1990). In this study, using DNase I footprinting, mobility shift assays, and mutagenesis, we demonstrate that at least three subdomains of this region are necessary to observe a full response to IL-6. The most distal contains a CTGGGAA motif, and its mutation inhibits IL-6 stimulation. Another, which is able to interact with several distinct nuclear proteins, among them members of the C/EBP family, is dispensable for IL-6 induction but plays an important role in the constitutive expression of beta Fg. Finally, a proximal hepatocyte nuclear factor 1 binding site, already described as the major determinant of beta Fg tissue-specific expression, is also required for IL-6 stimulation. These results indicate a complex interplay between nuclear proteins within the beta Fg IL-6-responsive region and suggest a tight functional coupling between the tissue-specific and inducible elements. PMID- 8423787 TI - p62cdc23 of Saccharomyces cerevisiae: a nuclear tetratricopeptide repeat protein with two mutable domains. AB - CDC23 is required in Saccharomyces cerevisiae for cell cycle progression through the G2/M transition. The CDC23 gene product contains tandem, imperfect repeats, termed tetratricopeptide repeats, (TPR) units common to a protein family that includes several other nuclear division CDC genes. In this report we have used mutagenesis to probe the functional significance of the TPR units within CDC23. Analysis of truncated derivatives indicates that the TPR block of CDC23 is necessary for the function or stability of the polypeptide. In-frame deletion of a single TPR unit within the repeat block proved sufficient to inactivate CDC23 in vivo, though this allele could rescue the temperature-sensitive defect of a cdc23 point mutant by intragenic complementation. By both in vitro and in vivo mutagenesis techniques, 17 thermolabile cdc23 alleles were produced and examined. Fourteen alleles contained single amino acid changes that were found to cluster within two distinct mutable domains, one of which encompasses the most canonical TPR unit found in CDC23. In addition, we have characterized CDC23 as a 62-kDa protein (p62cdc23) that is localized to the yeast nucleus. Our mutagenesis results suggest that TPR blocks form an essential domain within members of the TPR family. PMID- 8423786 TI - Sequences within the conserved cyclin box of human cyclin A are sufficient for binding to and activation of cdc2 kinase. AB - Cyclins are pivotal in the coordinate regulation of the cell cycle. By physical association, they are able to activate at least one of the cyclin-dependent kinases, cdc2. How this association between the catalytic moiety and cyclins leads to subsequent activation of the kinase remains unclear. In this report, we describe experiments to investigate this event at a physical level. Our approach was to map the regions required on the cyclin A molecule for interaction with cdc2. We have mapped the contact regions to two small noncontiguous stretches of amino acids, residues 189 to 241 and 275 to 320, both located within the conserved cyclin box domain of the protein. We have further shown that this region not only represents a contact site for cdc2 but apparently represents an intact functional domain with respect to cdc2 activation. This region alone is sufficient to stimulate maturation when injected into immature Xenopus laevis oocytes. This observation implies that events leading to the activation of cdc2 kinase can be mediated through small regions of the cyclin molecule that are located in the cyclin box. These regions contain some of the most highly conserved residues found between all the cyclin members so far identified. This suggests that the cyclin family members may have conserved a similar mechanism to bind and activate cyclin-dependent kinases. PMID- 8423788 TI - A new serum-responsive, cardiac tissue-specific transcription factor that recognizes the MEF-2 site in the myosin light chain-2 promoter. AB - We have identified a serum-responsive, cardiac tissue-specific transcription factor, BBF-1, that recognizes an AT-rich sequence (element B), identical to the myocyte enhancer factor (MEF-2) target site, in the cardiac myosin light chain-2 (MLC-2) promoter. Deletion of the element B sequence alone from the cardiac MLC-2 promoter causes, as does that of the MEF-2 site from other promoters and the enhancer of skeletal muscle genes, a marked reduction of transcription. BBF-1 is distinguishable from cardiac MEF-2 on the basis of immunoprecipitation with an antibody which recognizes MEF-2 but not BBF-1. Unlike MEF-2, BBF-1 is present exclusively in nuclear extracts from cardiac muscle cells cultured in a medium containing a high concentration of serum. Removal of serum from culture medium abolishes BBF-1 activity selectively with a concomitant loss of the positive regulatory effect of element B on MLC-2 gene transcription, indicating that there is a correlation between the BBF-1 binding activity and the tissue-specific role of the element B (MEF-2 site) sequence. The loss of element B-mediated activation of transcription is reversed following the refeeding of cells with serum containing medium. These data demonstrate that cardiac muscle cells contain two distinct protein factors, MEF-2 and BBF-1, which bind to the same target site but that, unlike MEF-2, BBF-1 is serum inducible and cardiac tissue specific. BBF-1 thus appears to be a crucial member of the MEF-2 family of proteins which will serve as an important tool in understanding the regulatory mechanism(s) underlying cardiogenic differentiation. PMID- 8423789 TI - In vivo and in vitro analysis of transcriptional activation mediated by the human cytomegalovirus major immediate-early proteins. AB - To define mechanistically how the human cytomegalovirus (HCMV) major immediate early (IE) proteins induce early-gene transcription, the IE1 72-kDa protein, the IE2 55-kDa protein, and the IE2 86-kDa protein were analyzed for their ability to activate transcription from an HCMV early promoter in vivo and in vitro. In transient-expression assays in U373MG astrocytoma/glioblastoma and HeLa cells, only the IE2 86-kDa protein was able to activate the HCMV early promoter to high levels. In HeLa cells, the IE1 72-kDa protein was able to activate the promoter to a low but detectable level, and the level of promoter activity observed in response to the IE2 86-kDa protein was increased synergistically following cotransfection of the constructs expressing both IE proteins. To examine the interaction of the HCMV IE proteins with the RNA polymerase II transcription machinery, we assayed the ability of Escherichia coli-synthesized proteins to activate the HCMV early promoter in nuclear extracts prepared from U373MG cells, HeLa cells, and Drosophila embryos. The results of the in vitro experiments correlated well with those obtained in vivo. The basal activity of the promoter was minimal in both the HeLa and U373MG extracts but was stimulated 6- to 10-fold by the IE2 86-kDa protein. With a histone H1-deficient extract from Drosophila embryos, the HCMV early promoter was quite active and was stimulated two- to fourfold by the IE2 86-kDa protein. Addition of histone H1 at 1 molecule per 40 to 50 bp of DNA template significantly repressed basal transcription from this promoter. However, the IE2 86-kDa protein, but none of the other IE proteins, was able to counteract the H1-mediated repression and stimulate transcription at least 10- to 20-fold. The promoter specificity of the activation was demonstrated by the inability of the IE2 86-kDa protein to activate the Drosophila Kruppel promoter in either the presence or absence of histone H1. These results suggest that one mechanism of transcription activation by the IE2 86-kDa protein involves antirepression. PMID- 8423790 TI - Characterization of the inducer of short transcripts, a human immunodeficiency virus type 1 transcriptional element that activates the synthesis of short RNAs. AB - The inducer of short transcripts, or IST, is an unusual transcriptional element located downstream of the human immunodeficiency virus type 1 (HIV-1) promoter. IST activates HIV-1 transcription, but the resulting RNAs are short and end at approximately position +59. IST, therefore, appears to promote the formation of transcription complexes that are unable to elongate efficiently. This activity contrasts with that of TAR, the target for Tat trans-activation, which upon binding of the viral protein Tat promotes the formation of transcription complexes capable of efficient elongation through the entire viral genome. We have localized and characterized the IST element. Our results indicate that IST is located mainly between positions -5 and +26, although the sequences from positions +40 to +59 also contribute to IST activity. Unlike TAR, which is an RNA element, IST appears to be a DNA element. Thus, the HIV-1 R region is a complex regulatory region with RNA and DNA elements that promote the formation of transcription complexes with different elongation properties. PMID- 8423791 TI - Different regulatory sequences control creatine kinase-M gene expression in directly injected skeletal and cardiac muscle. AB - Regulatory sequences of the M isozyme of the creatine kinase (MCK) gene have been extensively mapped in skeletal muscle, but little is known about the sequences that control cardiac-specific expression. The promoter and enhancer sequences required for MCK gene expression were assayed by the direct injection of plasmid DNA constructs into adult rat cardiac and skeletal muscle. A 700-nucleotide fragment containing the enhancer and promoter of the rabbit MCK gene activated the expression of a downstream reporter gene in both muscle tissues. Deletion of the enhancer significantly decreased expression in skeletal muscle but had no detectable effect on expression in cardiac muscle. Further deletions revealed a CArG sequence motif at position -179 within the promoter that was essential for cardiac-specific expression. The CArG element of the MCK promoter bound to the recombinant serum response factor and YY1, transcription factors which control expression from structurally similar elements in the skeletal actin and c-fos promoters. MCK-CArG-binding activities that were similar or identical to serum response factor and YY1 were also detected in extracts from adult cardiac muscle. These data suggest that the MCK gene is controlled by different regulatory programs in adult cardiac and skeletal muscle. PMID- 8423792 TI - Human biliary glycoprotein gene: characterization of a family of novel alternatively spliced RNAs and their expressed proteins. AB - Eight different human biliary glycoprotein (BGP) isoantigens, structurally related members of the carcinoembryonic antigen family, CD66/67 family, and immunoglobulin superfamily, are derived by alternative splicing from a single genomic transcription unit. Novel BGP isoforms have been identified by polymerase chain reaction amplification and by DNA sequencing of amplified cDNA segments. In addition to verifying previously documented BGPs, we describe four new forms, two of which have unusual nonimmunoglobulin exons contributed by inverted Alu repeats. Determination of the genomic DNA sequence encompassing most of the known extracellular and intracellular domains demonstrates that the translatable Alu like sequences are encoded in bona fide exons. The third novel BGP isoform contains none of the extracellular disulfide-linked immunoglobulin-like domains typical of these molecules but retains N-terminal and intracellular domains, suggesting distinct functions for N-terminal versus other disulfide-linked domains. cDNAs coding for each identified isoform have been transfected into COS7 monkey cells, and the resulting polypeptides are heavily N glycosylated but can be deglycosylated to their expected primary sizes. Many of these deglycosylated forms can be correlated with unique patterns of BGP expression in different cell lines, while in granulocytes, some previously undescribed or alternatively modified forms may predominate. The BGP family represents a potentially large but unknown source of functional diversity among cells of epithelial and hematopoietic origin. The availability of a defined set of expressed of BGP cDNAs should permit critical definition of their function. PMID- 8423793 TI - The rRNA enhancer regulates rRNA transcription in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, the rRNA genes are organized as a tandem array of head-to-tail repeats. An enhancer of rRNA transcription is present just at the end of each transcription unit, 2 kb away from the next one. This enhancer is unusual for S. cerevisiae in that it acts both upstream and downstream of, and even across, genes. The role of the enhancer in the nutritional regulation of rRNA transcription was studied by introducing a centromere plasmid carrying two rRNA minigenes in tandem, flanking a single enhancer, into cells. Analysis of the transcripts from the two minigenes showed that the enhancer was absolutely required for the stimulation of transcription of rRNA that occurs when cells are shifted from a poor carbon source to a good carbon source. While full enhancer function is provided by a 45-bp region at the 3' end of the 190-bp enhancer, some activity was also conferred by other elements, including both a T-rich stretch and a region containing the binding sites for the proteins Reb1p and Abf1p. We conclude that the enhancer is composed of redundant elements and that it is a major element in the regulation of rRNA transcription. PMID- 8423794 TI - Inhibition of protein kinase C zeta subspecies blocks the activation of an NF kappa B-like activity in Xenopus laevis oocytes. AB - Nuclear factor kappa B (NF-kappa B) plays a critical role in the regulation of a large variety of cellular genes. However, the mechanism whereby this nuclear factor is activated remains to be determined. In this report, we present evidence that in oocytes from Xenopus laevis, (i) ras p21- and phospholipase C (PLC) mediated phosphatidylcholine (PC) hydrolysis activates NF-kappa B and (ii) protein kinase C zeta subspecies is involved in the activation of NF-kappa B in response to insulin/ras p21/PC-PLC. Thus, the microinjection of either ras p21 or PC-PLC, or the exposure of oocytes to insulin, promotes a significant translocation to the nucleus of an NF-kappa B-like activity. This effect is not observed when oocytes are incubated with phorbol myristate acetate or progesterone, both of which utilize a ras p21-independent pathway for oocyte activation. These data strongly suggest a critical role of the insulin/ras p21/PC PLC/protein kinase C zeta pathway in the control of NF-kappa B activation. PMID- 8423795 TI - Transcription elongation in the human c-myc gene is governed by overall transcription initiation levels in Xenopus oocytes. AB - Both transcription initiation and transcription elongation contribute to the regulation of steady-state c-myc RNA levels. We have used the Xenopus oocyte transcription assay to study premature transcription termination which occurs in the first exon and intron of the human c-myc gene. Previous studies showed that after injection into Xenopus oocytes transcription from the c-myc P1 promoter resulted in read-through transcripts whereas transcription from the stronger P2 promoter resulted in a combination of prematurely terminated and read-through transcripts. We now demonstrate that this promoter-specific processivity results from the overall amount of RNA polymerase II transcription occurring from either promoter. Parameters that reduce the amount of transcription from P1 or P2, such as decreased concentration of template injected or decreased incubation time, result in a reduction in the ratio of terminated to read-through c-myc transcripts. Conversely, when transcription levels are increased by higher concentrations of injected template, increased incubation time, or coinjection with competing template, the ratio of terminated to read-through transcripts increases. We hypothesize that an RNA polymerase II processivity function is depleted above a threshold level of transcription initiation, resulting in high levels of premature transcription termination. These findings account for the promoter-specific effects on transcription elongation previously seen in this assay system and suggest a mechanism whereby limiting transcription elongation factors may contribute to transcription regulation in other eukaryotic cells. PMID- 8423796 TI - An essential yeast gene encoding a TTAGGG repeat-binding protein. AB - A yeast gene encoding a DNA-binding protein that recognizes the telomeric repeat sequence TTAGGG found in multicellular eukaryotes was identified by screening a lambda gt11 expression library with a radiolabeled TTAGGG multimer. This gene, which we refer to as TBF1 (TTAGGG repeat-binding factor 1), encodes a polypeptide with a predicted molecular mass of 63 kDa. The TBF1 protein, produced in vitro by transcription and translation of the cloned gene, binds to (TTAGGG)n probes and to a yeast telomeric junction sequence that contains two copies of the sequence TTAGGG separated by 5 bp. TBF1 appears to be identical to a previously described yeast TTAGGG-repeat binding activity called TBF alpha. TBF1 produced in vitro yields protein-DNA complexes with (TTAGGG)n probes that have mobilities on native polyacrylamide gels identical to those produced by partially purified TBF alpha from yeast cells. Furthermore, when extracts are prepared from a strain containing a TBF1 gene with an antigen tag, we find that the antigen copurifies with the predominant (TTAGGG)n-binding activity in the extracts. The DNA sequence of TBF1 was determined. The predicted protein sequence suggests that TBF1 may contain a nucleotide-binding domain, but no significant similarities to any other known proteins were identified, nor was an obvious DNA-binding motif apparent. Diploid cells heterozygous for a tbf1::URA3 insertion mutation are viable but upon sporulation give rise to tetrads with only two viable spores, both of which are Ura-, indicating that the TBF1 gene is essential for growth. Possible functions of TBF1 (TFB alpha) are discussed in light of these new results. PMID- 8423797 TI - Conditional silencing: the HMRE mating-type silencer exerts a rapidly reversible position effect on the yeast HSP82 heat shock gene. AB - The HMRE silencer of Saccharomyces cerevisiae has been previously shown to transcriptionally repress class II and class III genes integrated within the HMR silent mating-type locus up to 2.6 kb away. Here we study the ability of this element to repress at an ectopic position, independent of sequences normally associated with it. When integrated 750 bp upstream of the HSP82 heat shock gene, the silencer represses basal-level transcription approximately 5-fold but has no effect on chemical- or heat-shock-induced expression. Such conditional silencing is also seen when the HMRE/HSP82 allele is carried on a centromeric episome or when the entire HMRa domain is transplaced 2.7 kb upstream of HSP82. Notably, the a1 promoter within the immigrant HMRa locus remains fully repressed at the same time HSP82 is derepressed. The position effect mediated by the E silencer is absolutely dependent on the presence of a functional SIR4 gene product, is lost within 1 min following stress induction, and is fully reestablished within 15 min following a return to nonstressful conditions. Similar kinetics of reestablishment are seen in HMRE/HSP82 and HMRa/HSP82 strains, indicating that complete repression can be mediated over thousands of base pairs within minutes. DNase I chromatin mapping reveals that the ABF1, RAP1, and autonomously replicating sequence factor binding sites within the silencer are constitutively occupied in chromatin, unaltered by heat shock or the presence of SIR4. Similarly, the heat shock factor binding site upstream of HSP82 remains occupied under such conditions, suggesting concurrent occupancy of silencer and activator binding sites. Our results are consistent with a model in which silencing at the HMRE/HSP82 allele is mediated by direct or indirect contacts between the silencer protein complex and heat shock factor. PMID- 8423798 TI - Fusion of the erythropoietin receptor and the Friend spleen focus-forming virus gp55 glycoprotein transforms a factor-dependent hematopoietic cell line. AB - The Friend spleen focus-forming virus (SFFV) gp55 glycoprotein binds to the erythropoietin receptor (EPO-R), causing constitutive receptor signaling and the first stage of Friend erythroleukemia. We have used three independent strategies to further define this transforming molecular interaction. First, using a retroviral selection strategy, we have isolated the cDNAs encoding three fusion polypeptides containing regions of both EPO-R and gp55. These fusion proteins, like full-length gp55, transformed the Ba/F3 factor-dependent hematopoietic cell line and localized the transforming activity of gp55 to its transmembrane domain. Second, we have isolated a mutant of gp55 (F-gp55-M1) which binds, but fails to activate, EPO-R. We have compared the transforming activity of this gp55 mutant with the EPO-R-gp55 fusion proteins and with other variants of gp55, including wild-type polycythemia Friend gp55 and Rauscher gp55. All of the fusion polypeptides and mutant gp55 polypeptides were expressed at comparable levels, and all coimmunoprecipitated with wild-type EPO-R, but only the Friend gp55 and the EPO-R-gp55 fusion proteins constitutively activated wild-type EPO-R. Third, we have examined the specificity of the EPO-R-gp55 interaction by comparing the differential activation of murine and human EPO-R by gp55. Wild-type gp55 had a highly specific interaction with murine EPO-R; gp55 bound, but did not activate, human EPO-R. PMID- 8423799 TI - Heat shock factor is required for growth at normal temperatures in the fission yeast Schizosaccharomyces pombe. AB - Schizosaccharomyces pombe is becoming an increasingly useful organism for the study of cellular processes, since in certain respects, such as the cell cycle and splicing, it is similar to metazoans. Previous biochemical studies have shown that the DNA binding ability of S. pombe heat shock factor (HSF) is fully induced only under stressed conditions, in a manner similar to that of Drosophila melanogaster and humans but differing from the constitutive binding by HSF in the budding yeasts. We report the isolation of the cDNA and gene for the HSF from S. pombe. S. pombe HSF has a domain structure that is more closely related to the structure of human and D. melanogaster HSFs than to the structure of the budding yeast HSFs, further arguing that regulation of HSF in S. pombe is likely to reflect regulation in metazoans. Surprisingly, the S. pombe HSF gene is required for growth at normal temperatures. We show that the S. pombe HSF gene can be replaced by the D. melanogaster HSF gene and that strains containing either of these genes behave similarly to transiently heat-shocked strains with respect to viability and the level of heat-induced transcripts from heat shock promoters. Strains containing the D. melanogaster HSF gene, however, have lower growth rates and show altered morphology at normal growth temperatures. These data demonstrate the functional conservation of domains of HSF that are required for response to heat shock. They further suggest a general role for HSF in growth of eukaryotic cells under normal (nonstressed) growth conditions. PMID- 8423800 TI - Expression cDNA cloning of a transforming gene encoding the wild-type G alpha 12 gene product. AB - Using an expression cDNA cloning approach, we examined human tumor cell lines for novel oncogenes that might evade detection by conventional techniques. We isolated a transforming sequence that was highly efficient in transforming NIH 3T3 mouse fibroblasts. DNA sequence analysis identified the gene as the human homolog of a recently cloned alpha subunit of mouse GTP-binding protein G alpha 12. NIH 3T3 cells transfected with G alpha 12 cDNA grew in soft agar and were tumorigenic in nude mice. There were no apparent mutations in the cloned cDNA in comparison with a G alpha 12 cDNA clone isolated from a normal human epithelial cell library, implying that overexpression alone was sufficient to cause NIH 3T3 cell transformation. The observed altered growth properties mediated by G alpha 12 showed a certain degree of dependency on serum factors, and its mitogenic potential was also potently inhibited by suramin treatment. PMID- 8423801 TI - Autonomous expression of a noncatalytic domain of the focal adhesion-associated protein tyrosine kinase pp125FAK. AB - Integrins play a central role in cellular adhesion and anchorage of the cytoskeleton and participate in the generation of intracellular signals, including tyrosine phosphorylation. We have recently isolated a cDNA encoding a unique, focal adhesion-associated protein tyrosine kinase (FAK) that is a component of an integrin-mediated signal transduction pathway. Here we report the isolation of cDNAs encoding the C-terminal, noncatalytic domain of the FAK kinase, termed FRNK (FAK-related nonkinase). Both the FAK- and FRNK-encoded polypeptides, pp125FAK and p41/p43FRNK, are expressed in normal chicken embryo cells. pp125FAK and p41/p43FRNK were localized to focal adhesions, suggesting that pp125FAK is directed to the focal adhesions by sequences within its C terminal domain. We also show that the fibronectin-dependent increase in tyrosine phosphorylation of pp125FAK is accompanied by a concomitant posttranslational modification of p41FRNK. PMID- 8423802 TI - A new transcriptional-activation motif restricted to a class of helix-loop-helix proteins is functionally conserved in both yeast and mammalian cells. AB - Previous studies demonstrated that the amino-terminal portions of E2A and E2-2 are crucial for transactivation. Subsequent findings showed that the same amino terminal region of E2A is involved in two different translocation events contributing to the induction of a pre-B-cell acute lymphoblastic leukemia and a pro-B-cell acute lymphoblastic leukemia. These results led us to focus on the amino-terminal region of E2A to better understand its normal role in transcriptional regulation and its aberrant involvement in the two leukemias. We report here the identification of two conserved boxes in the E2A amino-terminal domain that show extensive homology within the transactivation domains of E12, E47, E2-2, HEB, and daughterless, all members of the same class of helix-loop helix proteins. Together, both boxes are crucial for transcriptional activation and have the potential to form a new activation motif, that of a loop adjacent to an amphipathic alpha-helix, designated the loop-helix (LH) motif. A minimal region containing the LH motif is sufficient for transcriptional activation. Point mutations in the amphipathic helix of the minimal region reduce its transactivation capabilities dramatically. The same constructs expressed in yeast cells show identical patterns of activation, suggesting that the LH motif and its target proteins are functionally conserved in yeast cells. We propose that the LH motif represents a novel transactivation domain that is distinct from the previously characterized acidic blob, proline-rich, and glutamine-rich activation motifs. In addition, the LH motif is the first activation motif restricted to one class of DNA binding proteins. PMID- 8423803 TI - Phosphorylation of the TAL1 oncoprotein by the extracellular-signal-regulated protein kinase ERK1. AB - Alteration of the TAL1 gene is the most common genetic lesion found in T-cell acute lymphoblastic leukemia. TAL1 encodes phosphoproteins, pp42TAL1 and pp22TAL1, that represent phosphorylated versions of the full-length (residues 1 to 331) and truncated (residues 176 to 331) TAL1 gene products, respectively. Both proteins contain the basic helix-loop-helix motif, a DNA-binding and protein dimerization motif common to several known transcriptional regulatory factors. We now report that serine residue 122 (S122) is a major phosphorylation site of pp42TAL1 in leukemic cell lines and transfected COS1 cells. In vivo phosphorylation of S122 is induced by epidermal growth factor with a rapid time course that parallels activation of the ERK/MAP2 protein kinases. Moreover, S122 is readily phosphorylated in vitro by the extracellular signal-regulated protein kinase ERK1. These data suggest that TAL1 residue S122 serves as an in vivo substrate for ERK/MAP2 kinases such as ERK1. Therefore, S122 phosphorylation may provide a mechanism whereby the properties of TAL1 polypeptides can be modulated by extracellular stimuli. PMID- 8423804 TI - Germ line maintenance of the pseudogene donor pool for somatic immunoglobulin gene conversion in chickens. AB - Somatic immunoglobulin diversity is generated in avian species by sequential gene conversion of variable (V) gene segments of the immunoglobulin heavy- and light chain loci during B-cell development. The germ line pools of donor sequence information for somatic V-region gene conversion are found in families of V pseudogenes, located 5' of the single functional V gene of each locus. The sequence relationships among the pseudogenes (psi VL) and functional VL1 gene of the chicken light-chain alleles in three inbred strains were compared to determine the extent of diversity within the germ line pseudogene cluster. Numerous differences were observed. For example, compared with the previously reported CB allele and the G4 allele, the S3 allele contains two intact pseudogenes between psi VL16 and psi VL18. These two adjacent psi VL gene segments (psi VL17a and psi VL17b) could have given rise to the psi VL17 segment of the G4 and CB alleles by homologous recombination. The majority of other sequence polymorphisms among the psi VL alleles appear to be the result of meiotic gene conversion. The incidence of untemplated mutations within psi VL segments is significantly lower than the incidence of mutation within the pseudogene flanking regions. Together with the observations that most psi VL segments have open reading frames and lack stop codons, these data support the hypothesis that the psi VL cluster resembles a functional multigene family maintained by evolutionary selection for its functional role in generating somatic antibody diversity. Meiotic gene conversion events within the psi VL cluster serve both to introduce diversity by the exchange of short segments between family members and to prevent the accumulation of random mutations. PMID- 8423805 TI - TSF3, a global regulatory protein that silences transcription of yeast GAL genes, also mediates repression by alpha 2 repressor and is identical to SIN4. AB - TSF3 encodes one of six (TSF1 to TSF6) recently identified global negative regulators of transcription in Saccharomyces cerevisiae. Mutant tsf3 strains exhibit defects in transcriptional silencing of the GAL1 promoter, allow expression from upstream activation sequence-less promoters, and exhibit pleiotropic defects in cell growth and development. Here we show that TSF3 is involved in transcriptional silencing mediated by the alpha 2 repressor and demonstrate that specific systems of transcriptional silencing may depend on the more global role of TSF3. Cloning and sequencing of TSF3 allowed us to predict a 974-amino-acid gene product identical to SIN4, a negative regulator of transcription of the HO (homothallism) mating type switching endonuclease. TSF3 disruptions are not lethal but result in phenotypes similar to those of the originally isolated alleles. Our results, together with those of Y. W. Jiang and D. J. Stillman (Mol. Cell. Biol. 12:4503-4514, 1992), suggest that TSF3 (SIN4) affects the function of the basal transcription apparatus, and this effect in turn alters the manner in which the latter responds to upstream regulatory proteins. PMID- 8423806 TI - Proto-oncogenes of the fos/jun family of transcription factors are positive regulators of myeloid differentiation. AB - The proto-oncogenes c-jun, junB, junD, and c-fos recently have been shown to encode for transcription factors with a leucine zipper that mediates dimerization to constitute active transcription factors; juns were shown to dimerize with each other and with c-fos, whereas fos was shown to dimerize only with juns. After birth, hematopoietic cells of the myeloid lineage, and some other terminally differentiated cell types, express high levels of c-fos. Still, the role of fos/jun transcription factors in normal myelopoiesis or in leukemogenesis has not been established. Recently, c-jun, junB, and junD were identified as myeloid differentiation primary response genes stably expressed following induction of terminal differentiation of myeloblastic leukemia M1 cells. Intriguingly, c-fos, though induced during normal myelopoiesis, was not induced upon M1 differentiation. To gain further insights into the role of fos/jun in normal myelopoiesis and leukemogenicity, M1fos and M1junB cell lines, which constitutively express c-fos and junB, respectively, were established. It was shown that enforced expression of c-fos, and to a lesser extent junB, in M1 cells results in both an increased propensity to differentiate and a reduction in the aggressiveness of the M1 leukemic phenotype. M1fos cells constitutively expressed immediate-early and late genetic markers of differentiated M1 cells. The in vitro differentiation of normal myeloblasts into mature macrophages and granulocytes, as well as the increased propensity of M1fos leukemic myeloblasts to be induced for terminal differentiation, was dramatically impaired with use of c-fos antisense oligomers in the culture media. Taken together, these observations show that the proto-oncogenes which encode for fos/jun transcription factors play important roles in promoting myeloid differentiation. The ability of the M1 leukemic myeloblasts to be induced for terminal differentiation in the absence of apparent fos expression indicates that there is some redundancy among the fos/jun family of transcription factors in promoting myeloid differentiation; however, juns alone cannot completely compensate for the lack of fos. Thus, genetic lesions affecting fos/jun expression may play a role in the development of "preleukemic" myelodysplastic syndromes and their further progression to leukemias. PMID- 8423807 TI - N-terminal DNA-binding domains contribute to differential DNA-binding specificities of NF-kappa B p50 and p65. AB - We previously reported that either oxidation or alkylation of NF-kappa B in vitro abrogates DNA binding. We used this phenomenon to help elucidate structural determinants of NF-kappa B binding. We now demonstrate that Cys-62 of NF-kappa B p50 mediates the redox effect and lies within an N-terminal region required for DNA binding but not for dimerization. Several point mutations in this region confer a transdominant negative binding phenotype to p50. The region is highly conserved in all Rel family proteins, and we have determined that it is also critical for DNA binding of NF-kappa B p65. Replacement of the N-terminal region of p65 with the corresponding region from p50 changes its DNA-binding specificity towards that of p50. These data suggest that the N-terminal regions of p50 and p65 are critical for DNA binding and help determine the DNA-binding specificities of p50 and p65. We have defined within the N-terminal region a sequence motif, R(F/G)(R/K)YXCE, which is present in Rel family proteins and also in zinc finger proteins capable of binding to kappa B sites. The potential significance of this finding is discussed. PMID- 8423808 TI - Identification of a 60-kilodalton stress-related protein, p60, which interacts with hsp90 and hsp70. AB - Immunoaffinity purification of hsp90 from chick oviduct cytosol reveals two major proteins, hsp70 and a 60-kDa protein (p60), copurifying with hsp90. A similar result is obtained when hsp90 is immunoaffinity purified from chick liver and brain cytosols, avian fibroblasts, and rabbit reticulocyte lysate. This p60 is the same protein previously identified in certain assembly complexes of chick progesterone receptor generated in a cell-free reconstitution system. Tryptic and cyanogen bromide peptide fragments were generated from gel-purified p60, and partial N-terminal sequences were determined from eight peptides. The sequences show a striking similarity to the sequence of a 63-kDa human protein (IEF SSP 3521) whose abundance is increased in MRC-5 fibroblasts following simian virus 40 transformation. A monoclonal antibody was prepared against avian p60; Western immunoblot analysis showed that p60 was present in each of eight chick tissues examined and in each of the human, rat, rabbit, and Xenopus tissues tested. Immunoaffinity purifications from both chick oviduct cytosol and rabbit reticulocyte lysate using anti-p60 and anti-hsp70 monoclonal antibodies confirm that there is a relatively abundant complex in these extracts containing hsp90, hsp70, and p60. This complex appears to comprise an important functional unit in the assembly of progesterone receptor complexes. However, judging from the abundance and widespread occurrence of this multiprotein complex, hsp90, hsp70, and p60 probably function interactively in other systems as well. PMID- 8423809 TI - The promoter region of the yeast KAR2 (BiP) gene contains a regulatory domain that responds to the presence of unfolded proteins in the endoplasmic reticulum. AB - The endoplasmic reticulum (ER) of eukaryotic cells contains an abundant 78,000-Da protein (BiP) that is involved in the translocation, folding, and assembly of secretory and transmembrane proteins. In the yeast Saccharomyces cerevisiae, as in mammalian cells, BiP mRNA is synthesized at a high basal rate and is further induced by the presence of increased amounts of unfolded proteins in the ER. However, unlike mammalian BiP, yeast BiP is also induced severalfold by heat shock, albeit in a transient fashion. To identify the regulatory sequences that respond to these stimuli in the yeast KAR2 gene that encodes BiP, we have cloned a 1.3-kb segment of DNA from the region upstream of the sequences coding for BiP and fused it to a reporter gene, the Escherichia coli beta-galactosidase gene. Analysis of a series of progressive 5' truncations as well as internal deletions of the upstream sequence showed that the information required for accurate transcriptional regulation of the KAR2 gene in S. cerevisiae is contained within a approximately 230-bp XhoI-DraI fragment (nucleotides -245 to -9) and that this fragment contains at least two cis-acting elements, one (heat shock element [HSE]) responding to heat shock and the other (unfolded protein response element [UPR]) responding to the presence of unfolded proteins in the ER. The HSE and UPR elements are functionally independent of each other but work additively for maximum induction of the yeast KAR2 gene. Lying between these two elements is a GC-rich region that is similar in sequence to the consensus element for binding of the mammalian transcription factor Sp1 and that is involved in the basal expression of the KAR2 gene. Finally, we provide evidence suggesting that yeast cells monitor the concentration of free BiP in the ER and adjust the level of transcription of the KAR2 gene accordingly; this effect is mediated via the UPR element in the KAR2 promoter. PMID- 8423810 TI - Enhancer-dependent transcription of the epsilon-globin promoter requires promoter bound GATA-1 and enhancer-bound AP-1/NF-E2. AB - We analyzed epsilon-globin transcription in erythroid cells and in erythroid extracts to determine the requirements for enhancer-dependent expression of this gene. Mutations that abolished GATA-1 binding at a single position in the promoter prevented interaction with enhancers, whereas elimination of a second more distal promoter GATA-1 site had no effect. Deletion or mutation of the GATA 1 sites in either the human beta-globin locus control region DNase-hypersensitive site II enhancer or the chicken beta A/epsilon-globin enhancer did not diminish the ability of the enhancers to interact with the promoter. In contrast, mutation of the AP-1/NF-E2 sites in these enhancers resulted in elimination of enhancement. In vitro transcription of these constructs was promoter dependent and was not sensitive to abolition of GATA-1 binding in the promoter, consistent with the role of GATA-1 solely as a mediator of the enhancer effect. Thus, GATA-1 regulates the response of the epsilon-globin gene to enhancers through a specific site in the promoter and requires enhancer AP-1/NF-E2 binding to transduce the enhancer effect on transcription. PMID- 8423811 TI - Correction of a deletion mutant by gene targeting with an adenovirus vector. AB - The usefulness of adenovirus type 5 as a vector for homologous recombination was examined in CHO cells by using the adenine phosphoribosyltransferase (aprt) gene. Infection of a hemizygous CHO APRT- cell line containing a 3-bp deletion in exon 5 of the aprt gene with a recombinant adenovirus containing the wild-type gene resulted in restoration of the APRT+ phenotype at a frequency of 10(-5) to 10(-6) per infected cell. A relatively high frequency (approximately 6 to 20%) of the transductants appears to result from a homologous recombination event. The mutation on the chromosomal aprt gene is corrected in the homologous recombinants, and APRT expression is restored to a normal hemizygous level. Neither adenovirus nor exogenous promoter sequences are detected in the homologous recombinants. The remaining transductants result from random integration of the aprt gene with the adenovirus sequence. A number of adenovirus vectors containing different promoter sequences linked to the hamster aprt gene were constructed. A possible role for the promoter region in the homologous recombination event was indicated by the lack of homologous recombination in constructs lacking an active promoter. PMID- 8423812 TI - In vivo regulation of rRNA transcription occurs rapidly in nondividing and dividing Drosophila cells in response to a phorbol ester and serum. AB - The synthesis of ribosomes is an essential cellular process which requires the transcription of the rRNA genes by RNA polymerase I (Pol I). The regulation of rRNA synthesis is known to be coupled to growth regulation. In nongrowing, slowly growing, and rapidly growing Drosophila cells, exposure to the tumor-promoting phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) increases the synthesis of precursor and mature rRNAs. Using nuclear run-on assays, we show that TPA enhances transcription of the rRNA genes. These results suggest that TPA regulates expression of RNA genes transcribed by Pol I, irrespective of the growth state of the cells. In slowly dividing Drosophila cells, increasing the serum concentration rapidly alters the accumulation of rRNA by enhancing rDNA transcription within 1 h. Thus, TPA and serum are each able to rapidly regulate rRNA gene expression in Drosophila cells. These results indicate that the RNA Pol I transcription system can be regulated by agents which have previously been shown to effect specific genes transcribed by the RNA Pol II system. PMID- 8423813 TI - In vitro transcription of Drosophila rRNA genes shows stimulation by a phorbol ester and serum. AB - The phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) and serum both stimulate rapid increases in the transcription of Drosophila rRNA genes in vivo. Here we report that this stimulation is observed in in vitro transcription assays using nuclear extracts from cells treated with TPA or serum. Experiments in which extracts from TPA- or serum-treated cells were mixed with extracts from cells grown in serum-restricted medium showed that there was an increased RNA polymerase I (Pol I) activity present in the cell extracts from treated cells. We used a series of plasmids that had been deleted in the region 5' to the start site of rRNA transcription to determine which sequences were necessary to support the increased transcription seen in extracts from stimulated cells. DNA templates that contain sequences between -150 and +32 (with +1 as the Pol I transcription start site) show dramatic increases in transcription with TPA- and serum stimulated cell extracts; however, templates that contain 5' sequences to -60 or 43 show at most one-third of the stimulation level of transcription in nuclear extracts from treated cells in comparison with untreated cell extracts. The 5' deletion to -34 abolishes the stimulation effect and drops the basal-level transcription by 20-fold. These results indicate that the regulation of Pol I transcription in Drosophila cells by serum and TPA requires two DNA elements, sequences from -150 to -60 (upstream control element) and sequences from -43 to 34 (a portion of the core promoter. PMID- 8423814 TI - Orientation and topography of RNA polymerase III in transcription complexes. AB - A photo-cross-linking method has been used to map the subunits of Saccharomyces cerevisiae RNA polymerase (Pol) III with respect to DNA in binary (preinitiation) and ternary (RNA-elongating) transcription complexes. Transcription factor- and Pol III-containing complexes have been assembled on S. cerevisiae SUP4 tRNA(Tyr) gene probes containing the photoactive nucleotide 5-[N-(p-azidobenzoyl)-3 aminoallyl]-dUMP in different specified positions. Covalent DNA-protein linkages form upon irradiation of these complexes, and the Pol III subunits that are cross linked to individual positions in the SUP4 tRNA gene have been identified. RNA Pol III cross-linking has been shown to require the box B downstream promoter element of the tRNA gene and the presence of transcription factor TFIIIB. Further proof of specificity has been provided by demonstrating that particular Pol III subunits move out of the range of upstream-placed photoactive nucleotides, and that others move into the range of downstream-placed photoactive nucleotides, as a consequence of initiating and elongating RNA chains. Binding and specific placement of Pol III have also been shown to require both the B' and the B" components of TFIIIB. Nine Pol III subunits are cross-linked from different positions of the SUP4 tRNA gene's nontranscribed strand. In binary transcription complexes, the two largest Pol III subunits are accessible to photo-cross-linking over the entire stretch of the DNase I footprint. The 27- and 34-kDa Pol III subunits are also relatively extended along DNA; its upstream projection makes the 34-kDa subunit a candidate for interaction with TFIIIB, while the 27-kDa subunit is accessible to photo-cross-linking from the leading edge of the Pol III binding site. Several subunits, including the 82- and 53-kDa subunits in binary transcription complexes, are relatively localized in their accessibility to cross linking. Multiple Pol III subunits are accessible to specific cross-linking from a single photoactive nucleotide in the middle of the transcription bubble of an arrested ternary transcription complex. It is suggested that this precisely placed transcription complex comprises a dynamic ensemble of structural states rather than a single perfectly constrained entity. PMID- 8423815 TI - Transcriptional activation by simian virus 40 large T antigen: interactions with multiple components of the transcription complex. AB - Simian virus 40 (SV40) large T antigen is a potent transcriptional activator of both viral and cellular promoters. Within the SV40 late promoter, a specific upstream element necessary for T-antigen transcriptional activation is the binding site for transcription-enhancing factor 1 (TEF-1). The promoter structure necessary for T-antigen-mediated transcriptional activation appears to be simple. For example, a promoter consisting of upstream TEF-1 binding sites (or other factor-binding sites) and a downstream TATA or initiator element is efficiently activated. It has been demonstrated that transcriptional activation by T antigen does not require direct binding to the DNA; thus, the most direct effect that T antigen could have on these simple promoters would be through protein-protein interactions with either upstream-bound transcription factors, the basal transcription complex, or both. To determine whether such interactions occur, full-length T antigen or segments of it was fused to the glutathione-binding site (GST fusions) or to the Gal4 DNA-binding domain (amino acids 1 to 147) (Gal4 fusions). With the GST fusions, it was found that TEF-1 and the TATA-binding protein (TBP) bound different regions of T antigen. A GST fusion containing amino acids 5 to 172 (region T1) efficiently bound TBP. TEF-1 bound neither region T1 nor a region between amino acids 168 and 373 (region T2); however, it bound efficiently to the combined region (T5) containing amino acids 5 to 383.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8423816 TI - Increased UV resistance of a xeroderma pigmentosum revertant cell line is correlated with selective repair of the transcribed strand of an expressed gene. AB - A UV-resistant revertant (XP129) of a xeroderma pigmentosum group A cell line has been reported to be totally deficient in repair of cyclobutane pyrimidine dimers (CPDs) but proficient in repair of 6-4 photoproducts. This finding has been interpreted to mean that CPDs play no role in cell killing by UV. We have analyzed the fine structure of repair of CPDs in the dihydrofolate reductase gene in the revertant. In this essential, active gene, we observe that repair of the transcribed strand is as efficient as that in normal, repair-proficient human cells, but repair of the nontranscribed strand is not. Within 4 h after UV at 7.5 J/m2, over 50% of the CPDs were removed, and by 8 h, 80% of the CPDs were removed. In contrast, there was essentially no removal from the nontranscribed strand even by 24 h. Our results demonstrate that overall repair measurements can be misleading, and they support the hypothesis that removal of CPDs from the transcribed strands of expressed genes is essential for UV resistance. PMID- 8423817 TI - Acquisition of telomere repeat sequences by transfected DNA integrated at the site of a chromosome break. AB - Previous analysis of plasmid DNA transfected into 108 cell clones demonstrated extensive polymorphism near the integration site in one clone. This polymorphism was apparent by Southern blot analysis as diffuse bands that extended over 30 kb. In the present study, nucleotide sequence analysis of cloned DNA from the integration site revealed telomere repeat sequences at the ends of the integrated plasmid DNA. The telomere repeat sequences at one end were located at the junction between the plasmid and cell DNA. The telomere repeat sequences at the other end were located in the opposite orientation in the polymorphic region and were shown by digestion with BAL 31 to be at the end of the chromosome. Telomere repeat sequences were not found at this location in the plasmid or parent cell DNA. Although the repeat sequences may have been acquired by recombination, a more likely explanation is that they were added to the ends of the plasmid by telomerase before integration. Comparison of the cell DNA before and after integration revealed that a chromosome break had occurred at the integration site, which was shown by fluorescent in situ hybridization to be located near the telomere of chromosome 13. These results demonstrate that chromosome breakage and rearrangement can result in interstitial telomere repeat sequences within the human genome. These sequences could promote genomic instability, because short repeat sequences can be recombinational hotspots. The results also show that DNA rearrangements involving telomere repeat sequences can be associated with chromosome breaks. The introduction of telomere repeat sequences at spontaneous or ionizing radiation-induced DNA strand breaks may therefore also be a mechanism of chromosome fragmentation. PMID- 8423818 TI - Histone H2A.X gene transcription is regulated differently than transcription of other replication-linked histone genes. AB - Histone H2A.X is a replication-independent histone H2A isoprotein species that is encoded by a transcript alternatively processed at the 3' end to yield two mRNAs: a 0.6-kb mRNA ending with the stem-loop structure characteristic of the mRNAs for replication-linked histone species, and a second, polyadenylated 1.6-kb mRNA ending about 1 kb further downstream (C. Mannironi, W. M. Bonner, and C. L. Hatch, Nucleic Acids Res. 17:9113-9126, 1989). Of the two, the 0.6-kb H2A.X stem loop mRNA predominates in many cell lines, indicating that the presence of two types of mRNA may not completely account for the replication independence of H2A.X protein synthesis. The ambiguity is resolved by the finding that the level of the 0.6-kb H2A.X mRNA is only weakly downregulated during the inhibition of DNA replication and only weakly upregulated during the inhibition of protein synthesis, while the levels of other replication-linked mRNAs are strongly down- or upregulated under these two conditions. Analysis of the nuclear transcription rates of specific H2A genes showed that while the rates of transcription of replication-linked H2A genes decreased substantially during the inhibition of DNA synthesis and increased substantially during the inhibition of protein synthesis, the rate of H2A.X gene transcription decreased slightly under both conditions. These differences in transcriptional regulation between the H2A.X gene and other replication-linked histone genes are sufficient to account for the differences in regulation of their respective stem-loop mRNAs. PMID- 8423820 TI - Ca-tension relationships of muscle fibers from patients with periodic paralysis. PMID- 8423821 TI - Gangliosides attenuate vincristine neurotoxicity on dorsal root ganglion cells. AB - We investigated the effects of bovine brain gangliosides on the neurotoxicity of vincristine in dissociated cultures of dorsal root ganglion cells from 10-day chick embryos. The effects of the drugs were quantified as the numbers of neurite bearing cells or total neurite length in individual neurite-bearing cells. The administration of vincristine (1 to 1000 pg/mL) inhibited neurite outgrowth from the cells, whereas gangliosides (10 to 1000 micrograms/mL) protected them against this inhibition in a concentration-dependent manner. Electron microscopy revealed vincristine-induced fragmentation and longitudinal disorientation of microtubules in neurites and showed the protection by gangliosides against such damaging effects. Our results show that exogenous administration of gangliosides attenuates the neurotoxicity of vincristine in vitro. PMID- 8423819 TI - Combinatorial regulation by promoter and intron 1 regions of the metallothionein SpMTA gene in the sea urchin embryo. AB - The SpMTA metallothionein gene of the sea urchin Strongylocentrotus purpuratus is regulated developmentally, histospecifically, and by heavy-metal induction. The sequenced 5' flank of the gene can be divided into proximal, middle, and distal regions, each containing a pair of metal response elements (MREs). Canonical 7-bp core sequences are present in all except the middle-region MREs c and d, which contain 1-bp mismatches. Metal-induced expression in transgenic blastulae was increased with each consecutive addition of the middle and distal regions to a chimeric reporter gene construct containing the proximal SpMTA promoter region. Reduced metal induction through point mutation of the distal MREs e and f indicated that the MREs themselves were largely responsible for the transcriptional increase. These activities were further enhanced by SpMTA intron 1, but not when a specific interior region of the intron had been deleted. The atypical MREs c and d did not support induction by themselves, i.e., when present alone with mutated proximal MREs a and b. However, in the presence of intron 1, they were able to substitute for the nullified MREs a and b in the promotion of metal-induced expression. This capability suggests, furthermore, that these atypical MREs, in addition to responding to an intron 1 region, participate cooperatively with the canonical proximal MREs. PMID- 8423822 TI - Repetitive nerve stimulation versus twitch tension in rats with EAMG. PMID- 8423823 TI - Sural nerve biopsies in Guillain-Barre syndrome: axonal degeneration and macrophage-associated demyelination and absence of cytomegalovirus genome. PMID- 8423824 TI - The vitality of the sciatic nerve of the frog and rat in a chamber which allows maintained in vitro recording of the compound nerve action potentials. PMID- 8423825 TI - Autonomic function in HMSN: evidence of slowed sudomotor conduction? PMID- 8423826 TI - Atropine modulates presynaptic function in mice with passively transferred Lambert-Eaton myasthenic syndrome. PMID- 8423827 TI - Lymphocyte subsets in amyotrophic lateral sclerosis with motor conduction block. PMID- 8423828 TI - Needle electromyography of the diaphragm: a new technique. AB - Electrodiagnostic evaluation of diaphragmatic function has consisted of phrenic nerve stimulation and surface or esophageal recordings of the electrical activity of the diaphragm. Needle electromyography of the diaphragm has rarely been reported because of the perceived danger of this procedure. We describe a new technique for needle electromyography of the diaphragm. An EMG electrode is placed in the costal insertion of the diaphragm under the 8th, 9th, or 10th rib cartilage, distant from the major vessels, pleura, lungs, and abdominal viscera. Diaphragmatic denervation was found in 42 of 81 patients using this method. There were no complications related to the procedure. Needle electromyography of the diaphragm provides important information in the diagnosis and management of respiratory dysfunction. PMID- 8423829 TI - Duration of amyotrophic lateral sclerosis is age dependent. AB - Since 1985, we prospectively followed 246 patients with ALS. The relationship between the age of developing neurological impairment and disease duration was analyzed in 138 patients (86 men and 52 women) who died. Mean disease duration was 4.0 +/- 3.8 years for men and 3.2 +/- 2.5 years for women. There was an inverse, exponential, relationship between onset age and duration (goodness-of fit P > 0.05). Mean duration at onset age < or = 40 years was 8.2 +/- 5.0 years compared with 2.6 +/- 1.4 years for patients aged 61 to 70 years (P > 0.001). The ratio of young (< or = 40 years) men to women was 3.6:1. When matched for age, disease duration was the same for patients with bulbar and nonbulbar onsets. We conclude that onset age, but not sex, is the most significant predictor determining disease duration in ALS. Longer survival in younger patients probably reflects their greater neuronal reserve. PMID- 8423830 TI - Effect of caffeine and high potassium on normal and dystrophic mouse EDL muscles at various developmental stages. AB - EDL muscles from normal and dystrophic (dy2j) mice of various ages were examined. Muscles were divided into three groups according to age: 7 to 14 days postnatal, 16 to 21 days postnatal, and 6 months old, to assess age and/or phenotype related differences in the muscle response to caffeine or high K+. The response of normal muscles to caffeine decreased with age and reached adult characteristics between the second and third week of postnatal life. Their response to high K+ also changed during postnatal development; specifically, the time taken to recover to 50% pretest twitch tension decreased with age, probably reflecting developmental changes in Cl- conductance. Up to 21 days of age, the sensitivity of dystrophic muscles to both caffeine and high K+ was essentially similar to normal, while marked differences were observed in the adult. Taken altogether, our results suggest that while the maturation of a number of systems might be delayed in dystrophic muscles at preclinical stages of the disease, their e-c coupling and SR function (Ca2+ release and reuptake) appear to be quite normal. Our results further suggest that the "abnormal" responses of dystrophic muscles at more advanced stages of the disease, when challenged by drugs acting on either of these systems, may be explained in terms of changes in muscle fiber type proportions. PMID- 8423831 TI - An augmented computer model of motor unit reorganization in neurogenic diseases of skeletal muscle. AB - A computer model of denervation and complete reinnervation in skeletal muscle was originally developed for the purpose of furthering an understanding of the underlying mechanisms of motor unit reorganization in neurogenic diseases. We now describe its successor, a computer model for investigating different rates of denervation and reinnervation, as well as incomplete reinnervation. The new model introduces the concept of permanent denervation and features enhanced interactive control over the distribution of motor unit centers and additional measures of dispersion and co-dispersion of muscle fibers. The use of this model for investigating pathophysiologically significant issues in denervating diseases is illustrated with five different sets of parameters. These simulate some of the processes that may be operational in chronic spinal muscular atrophy, amyotrophic lateral sclerosis, and progressive postpolio muscular dystrophy. The enhanced model will allow in-depth analysis of the influence of hypothesized pathophysiological processes on clinical, electrophysiological and pathological outcomes in human disease. PMID- 8423832 TI - A case of myopathy associated with a dystrophin gene deletion and abnormal glycogen storage. AB - A 30-year-old man with no family history of muscle disease presented with a progressive proximal myopathy and calf hypertrophy characteristic of Becker muscular dystrophy. A deletion of exons 45 to 48 in the dystrophin gene was confirmed by Southern blotting and multiplex polymerase chain reaction. However, muscle biopsy showed massive accumulation of glycogen, although no significant abnormality of glycolytic pathway enzymes could be demonstrated. This patient therefore has a previously undescribed myopathy associated with both Becker muscular dystrophy and a glycogen storage disorder of unknown aetiology. PMID- 8423833 TI - The quadriceps strength of healthy elderly people remeasured after eight years. AB - Isometric quadriceps strength was remeasured in 14 healthy survivors of a group of elderly people first studied 8 years previously. There were 4 men (median age 81 years, range 79 to 84) and 10 women (82 years, range 79 to 89). They were selected for their health, not their levels of physical activity. Nevertheless, they were active when first studied and, with 1 exception, had maintained or increased their levels of physical activity. Isometric quadriceps strength was well preserved; the median change in the strength of the stronger quadriceps was only -0.3% per annum (95% confidence interval = -1.4 to +0.8). PMID- 8423835 TI - Non-reflex and reflex mediated ankle joint stiffness in multiple sclerosis patients with spasticity. AB - In this study, we have measured the passive, the intrinsic, and the reflex mediated mechanical response to stretch of the ankle extensors and flexors in 13 spastic multiple sclerosis patients and 10 healthy control subjects. In the ankle flexors, the patients had no reflex-mediated stiffness. The passive stiffness was increased by 138% (95% confidence interval: 26-91%) and the intrinsic stiffness by 79% (41-158%) when compared with the healthy subjects. In the ankle extensors, the reflex-mediated stiffness and the intrinsic stiffness of the patients were equal to the reflex-mediated and the intrinsic stiffness in healthy subjects. The passive stiffness was increased by 152% (41-352%). We conclude that spastic muscles in multiple sclerosis patients have an increased non-reflex stiffness (passive plus intrinsic stiffness), and that the reflex-mediated stiffness in the extensors during a sustained voluntary contraction does not differ significantly from healthy subjects. PMID- 8423834 TI - Progressive unilateral hypertrophic myopathy: a case study. AB - A 12-year-old girl had progressive unilateral muscle hypertrophy limited to the sole, tibialis anterior, and biceps femoris muscles. The affected muscles showed complex repetitive discharges by electromyography, necrosis and variation of fiber size upon histopathological examination, and increased metabolic activity in biochemical studies. The findings suggest a myopathic origin, but the actual cause remains unknown. PMID- 8423836 TI - Acute myopathy and neuropathy in status asthmaticus: case report and literature review. AB - A 38-year-old woman developed acute, severe weakness during the treatment of status asthmaticus that included high-dose intravenous corticosteroids. A muscle biopsy and EMG indicated a myopathy, and nerve conduction studies disclosed a neuropathic component. In association with corticosteroid tapering, the clinical, EMG, and nerve conduction abnormalities resolved. In some patients, intensive treatment of status asthmaticus may cause a reversible, toxic disorder of muscle and nerve. PMID- 8423837 TI - Dissociation of [H+] from fatigue in human muscle detected by high time resolution 31P-NMR. AB - Previous in vivo studies of skeletal muscle fatigue have demonstrated significant relationships between the decline of muscular force and changes in muscle metabolism. However, these studies performed measurements over relatively long time intervals or during steady state exercise, thereby obscuring rapid metabolic changes occurring at the onset of exercise and recovery. To overcome these limitations, fatigue of human calf musculature during sustained isometric foot plantar flexion was quantified continuously as the decline in maximal voluntary contraction force (MVC), while concentrations of phosphocreatine (PCr), inorganic phosphate (Pi), intracellular free hydrogen ion (H+), and monovalent phosphate (H2PO4-) were simultaneously measured at 2-second intervals by 31P nuclear magnetic resonance. The first major finding was that [H+], which has been thought to be a mediator of muscle fatigue, actually declined during the first 10 seconds of exercise when force was declining and rose immediately postexercise, when force partially recovered. Second, the correlations of [H+], [H2PO4-] and Pi with MVC during the first minute of exercise were determined to be curvilinear and not linear as previously suggested. Furthermore, using either a linear or curvilinear regression model, [H2PO4-] and Pi demonstrated a closer correlation to MVC than [H+] during the first minute of exercise. Thus, these results reveal nuances in the relationships of MVC to metabolites previously undetected by low time resolution measurements. These findings suggest that during sustained isometric exercise, rising [H+] is not likely to be the sole mechanism of muscle fatigue and are consistent with the view that a rise of Pi or [H2PO4-] is a major causation factor in force reduction. PMID- 8423838 TI - Distinguishing unloading- versus reloading-induced changes in rat soleus muscle. AB - Previously, solei from rats orbited 12.5 days aboard Cosmos 1887 biosatellite were biopsied 48-56 hours postflight. These atrophic muscles showed severe pathology. Designing a ground-based model of that space flight, we tested the hypothesis that 48 hours of postflight muscle reloading induced pathological changes. Rats were subjected to 12.5 days of hindlimb suspension unloading and biopsied immediately after suspension or after returning to normal weightbearing 12, 24, or 48 hours. Soleus morphological changes were quantitated on histochemically and immunohistochemically stained cross-sections. Solei from 0 hour reloaded rats showed significantly decreased wet weights, diminished myofiber cross-sectional area, angular myofibers, myofibril disruption, and more myofibers expressing fast myosin. Compared with suspension alone (0-hour reloading), reloading 12-48 hours induced slightly increased soleus wet weights, myofiber swelling, significant interstitial tissue edema, macrophage activation, and monocyte infiltration. These results suggest the degree and type of muscle degenerative changes observed postflight depend on the duration of gravity readaptation before biopsy and not solely on exposure to microgravity. PMID- 8423839 TI - Patent for a mouse. PMID- 8423841 TI - Healy, hundreds of other exit with Bush. PMID- 8423840 TI - Historic Bart's fights for its life under medical school reorganization. PMID- 8423842 TI - RAC defers to NIH director on some gene therapy cases. PMID- 8423843 TI - Mediation sought on biotechnology disputes. PMID- 8423844 TI - Procedure for gene patents. PMID- 8423845 TI - Body heat. PMID- 8423846 TI - Thatcher right? PMID- 8423847 TI - Mimicking ligands. PMID- 8423848 TI - Mimicking ligands. PMID- 8423849 TI - Long-range correlations in DNA. PMID- 8423850 TI - Early T-cell development. PMID- 8423851 TI - Vertical disparities, differential perspective and binocular stereopsis. AB - To calculate the depth difference between a pair of points on a three-dimensional surface from binocular disparities, it is necessary to know the absolute distance to the surface. Traditionally, it has been assumed that this information is derived from non-visual sources such as the vergence angle of the eyes. It has been shown that the horizontal gradient of vertical disparity between the images in the two eyes also contains information about the fixation distance. Recent results, however indicated that manipulations of the vertical disparity gradient have no effect on either the perceived shape or the perceived depth of surfaces defined by horizontal disparities. Following the reasoning of Longuet-Higgins and Tyler, we suggest that vertical disparities are best understood as a consequence of perspective viewing from two different vantage points and the results we report here show that the human visual system is able to exploit vertical disparities and use them to scale the perceived depth and size of stereoscopic surfaces, if the field of view is sufficiently large. PMID- 8423852 TI - Visible flicker from invisible patterns. AB - Using a laser interferometer we can create grating patterns of high optical contrast (interference fringes) directly on the retina. With coarse fringe patterns, the alternating light and dark bars of the pattern can be seen, but the bars of the finest fringes are not subjectively resolved. We report here that when we rapidly modulate the contrast of a fine fringe pattern (keeping overall luminance constant), observers experience flicker, even if the fringes are too finely spaced to be perceived as a grating. For this flicker to be seen, the pattern needs to be resolvable by the photoreceptors themselves, but not necessarily by later stages of visual processing. It can be explained if, in man, signals associated with individual cone receptors do not depend linearly on light intensity, but instead are scaled by a fast sensitivity-regulating or light adaptation mechanism. Contrast-modulation flicker is not demonstrable in rod vision; rod vision therefore lacks such a local adaptation process. PMID- 8423853 TI - Right-handed rotation of an actin filament in an in vitro motile system. AB - Muscle contraction occurs by mutual sliding between thick (myosin) and thin (actin) filaments. But the physical and chemical properties of the sliding force are not clear; even the precise direction of sliding force generated at each cross-bridge is not known. We report here the use of a recently developed in vitro motile assay system to show supercoiling of an actin filament in which the front part of the filament was fixed to a glass surface through cross-linked heavy-meromyosin and the rear part was able to slide on a track of heavy meromyosin. A left-handed single turn of superhelix formed just before supercoiling, suggesting that the sliding force has a right-handed torque component that induces the right-handed rotation of an actin filament around its long axis. The presence of the torque component in the sliding force will explain several properties of the contractile system of muscle. PMID- 8423854 TI - DNA polymerase-alpha is essential for mating-type switching in fission yeast. AB - In the fission yeast Schizosaccharomyces pombe, the double-stranded chromosomal break (DSB) at the mating-type locus (mat1) initiates recombination during mating type switching. A constant DSB level is maintained throughout the cell-cycle. In the strand-segregation model for mating-type switching, it was postulated that if the DSB is generated during or soon after mat1 replication, one of the chromatids could be repaired and switched during replication in the next cell cycle, while the other chromatid inherits the break. Here we report a molecular characterization of swi7, one of the genes required for DSB formation. Surprisingly, a gene complementing the swi7 mutation maps to chromosome I and encodes S. pombe DNA polymerase-alpha. Disruption of this gene is lethal in both switching and non-switching strains, as expected. S. pombe DNA polymerase-alpha must therefore play a role in generating the DSB at mat1, suggesting that DSB formation is coupled with DNA replication. PMID- 8423856 TI - Unusual clustering of carboxyl side chains in the core of iron-free ribonucleotide reductase. AB - The principal driving forces of protein folding are the burial of hydrophobic residues in the interior of proteins and the exposure of charged residues at the surface. Charged residues are only occasionally found in the interior, where they form hydrogen bonds to oppositely charged residues or main-chain atoms. Ribonucleotide reductase, a key enzyme in DNA synthesis, catalyses the de novo production of deoxyribonucleotide precursors. It is composed of two different dimeric proteins R1 and R2 (refs 3-5). R2 subunits contain buried iron-centres with each centre formed by two ferric ions coordinated by four carboxylates and two histidine ligands. Iron-free R2, apoR2, is a precursor of active R2 and folds into a stable protein which is transformed into active R2 by ferrous ions and molecular oxygen. Here we show that the iron-free protein does not undergo any major structural changes compared with the iron-containing R2. The effect of this is a clustering of four carboxyl side chains in the interior of the subunit, in contrast to the normal distribution of charged residues in proteins. PMID- 8423855 TI - Viral DNA and a viral peptide can act as cofactors of adenovirus virion proteinase activity. AB - Human adenovirus (Ad2), like many other viruses, contains a virion-associated proteinase essential for the synthesis of infectious virus particles. We observed proteinase activity in wild-type virus but not in the ts-1 virus, which contains a mutation in the Ad2 L3 endoprotease gene that confers temperature-sensitive processing of virion precursor proteins. Unexpectedly, we did not observe proteinase activity with purified recombinant endoprotease protein (M(r) 23 K). Purified recombinant endoprotease protein, however, complemented the mutation in ts-1 virions, restoring proteinase activity when mixed together. This implied that cofactors may be required. Here we reconstitute proteinase activity in vitro with three purified viral components: (1) the recombinant endoprotease protein; (2) an 11-amino-acid peptide that originates from the carboxy terminus of pVI, the precursor to virion component VI; and (3) adenovirus DNA. The use of DNA for a proteinase activity is unprecedented. PMID- 8423857 TI - [Numbness of the chin, sometimes an ominous sign]. PMID- 8423858 TI - [Balloon angioplasty and heart surgery; expectations and needs in the 90s]. PMID- 8423859 TI - [Cocaine and pregnancy; who pays the bill?]. PMID- 8423860 TI - [Controversies in preventive health care. III. Juvenile health care]. PMID- 8423861 TI - [Consensus therapy of sexually transmissible diseases]. AB - The organization and the management of sexually transmitted diseases shows large differences between general practitioners and specialists and also between specialists in dermato-venereology, obstetrics and urology. Changing insights in epidemiology, growing knowledge in complications, new technology in the diagnosis of sexually transmitted diseases and limited therapeutic possibilities have important implications for the different options in the control of sexually transmitted diseases. By promoting uniformity in strategies for sexually transmitted diseases these consensus guidelines have tried to formulate minimal standards for diagnostic procedures, therapy and follow-up for the most important sexually transmitted diseases. PMID- 8423862 TI - [The epidemiological transition in The Netherlands]. AB - OBJECTIVE: To assess the differences in the pattern of mortality by sex, age and cause of death between 1875-1879 and 1970 for the Netherlands, and the contribution of different causes of death to the decrease of total mortality between these periods. METHODS: Mortality data for 1875-1879 were extracted from one of the first publications after the start of the national cause-of-death register, and were reclassified according to the cause-of-death classification in use in 1970. The mortality data for 1875-79 and 1970 were directly standardised using the age structure of the Dutch population in 1930. RESULTS: The mortality rates in 1970 were only a third (for men) to a quarter (for women) of those in 1875-79. Mortality patterns by age changed drastically, due to stronger mortality reductions among younger people. The share in total mortality decreased enormously for infectious diseases (from 38-39% to 4-5%) and for mental disorders/diseases of the nervous system (9-11% to 2-3%), but the shares of cancer (2-3% to 25-26%), cardiovascular diseases (7-8% to 42%) and external causes (1-3% to 7-9%) increased correspondingly. Almost 50% of the decrease of total mortality was due to a number of infectious diseases, of which respiratory diseases were the most important subgroup. CONCLUSION: The Dutch population experienced an 'epidemiological transition' very much like that described for some other countries. PMID- 8423863 TI - [Bronchospasm, apnea and heart arrest following dipyridamole perfusion scintigraphy]. AB - The occurrence of fatal respiratory insufficiency following dipyridamole-thallium imaging is described. The patient, a 67-year-old man, had a history of chronic obstructive lung disease. Since patients with a history of chronic obstructive lung disease have an increased risk of developing bronchospasm after dipyridamole infusion, it is advised to be cautious in performing dipyridamole-thallium imaging in these patients. Dobutamine may be an acceptable alternative to dipyridamole in these patients. PMID- 8423864 TI - [Where does one perform laboratory studies in clinical-pharmaceutical research? Medical-ethical considerations]. PMID- 8423865 TI - [Increase in gonorrhea in younger homosexual men in Amsterdam]. PMID- 8423866 TI - [Letters from Washington. Food and Drug Administration and medical technology: at an impasse?]. PMID- 8423867 TI - [Medical residents across the borders: Nijmegen versus Boston]. PMID- 8423868 TI - Internal medicine training in Nijmegen and Boston: observations from an American trained physician. PMID- 8423869 TI - Neurosurgical horizons in Parkinson's disease. AB - Based on recent neuroanatomic and physiologic discoveries, neurosurgical therapies may increasingly complement and extend pharmacologic management of Parkinson's disease. Procedures showing promise include subthalamotomy and pallidotomy; thalamic electrical stimulation may also offer application for tremor control. Transplantation of adrenal chromaffin cells has not been associated with consistent long-term improvement in most patients, and fetal mesencephalic transplantation remains controversial. Trophic factors that may be pivotal to cellular repair and survival of transplanted tissue have potential therapeutic roles when purified and perfused centrally or when the cells that produce the factors are transplanted. PMID- 8423870 TI - A controlled trial with vasopressin analogue (DGAVP) on cognitive recovery immediately after head trauma. AB - Recent evidence suggests that the vasopressin analogue desglycinamide-arginine8 vasopressin (DGAVP) might specifically benefit mild brain trauma patients. We investigated the effect of intranasal DGAVP treatment in 32 patients who had sustained a mild head injury for 3 months in a double-blind, placebo-controlled, matched-pairs study. DGAVP did not have a positive effect on cognitive recovery in this group of mildly affected patients. PMID- 8423871 TI - Clock-drawing test and unilateral spatial neglect. AB - We investigated the ability of 25 patients with left unilateral spatial neglect to make a clock face by putting numbers inside a printed circle. Impairment seen in this clock-drawing test did not parallel neglect severity as judged by results of the line-cancellation and line-bisection tests, as well as the copying of a daisy. The score for clock drawing correlated highly with the verbal WAIS score. Most neglect patients with a verbal IQ of 87 or more could draw a clock face fairly well and used planning in placing the numbers 12, 3, 6, and 9 before the others. In clock drawing, verbal intelligence may compensate for left unilateral spatial neglect. We therefore recommend use of the line-cancellation and line bisection tests, as well as the copying test, but do not recommend use of the clock-drawing test in the diagnosis of left unilateral spatial neglect. PMID- 8423872 TI - The use of magnetic resonance imaging in diagnosing infantile neuroaxonal dystrophy. AB - We used MRI to investigate the brains of four children ranging from 3 to 10 years of age with infantile neuroaxonal dystrophy. T2-weighted imaging revealed characteristic findings of marked cerebellar atrophy and diffuse hyperintensity of the cerebellar cortex. At autopsy, one child had extensive astrogliosis and neuronal loss with shrinkage of the cerebellar cortex, in addition to typical widespread changes of neuroaxonal dystrophy. The characteristic hyperintensity of the cerebellar cortex on T2-weighted imaging probably is secondary to the extensive gliosis and shrinkage of the cerebellar cortex. These cerebellar findings on MRI may permit early diagnosis of infantile neuroaxonal dystrophy. PMID- 8423873 TI - A novel antineuronal antibody in stiff-man syndrome. AB - Two-thirds of stiff-man syndrome (SMS) patients harbor an autoantibody specific for a 64-kD species of glutamic acid decarboxylase (GAD), the rate-limiting enzyme in GABA synthesis. We assayed SMS antisera from two patients with SMS for the presence of anti-GAD antibodies using Western blot, immunohistochemical, and enzymatic analyses. Both SMS antisera recognized an 80-kD antigen present in human and rat neuronal extracts, and failed to recognize the 64-kD GAD species. Immunohistochemistry demonstrated neuronal binding identical to that reported with anti-GAD antibodies. Both sera depleted GAD activity from brain extracts. Our analysis indicates that these SMS antisera differ from previously reported SMS antisera by recognizing a novel 80-kD antigen, and suggests that they contain antibodies directed against either a species of GAD different in size from the 64 kD enzyme, or a protein that co-immunoprecipitates with GAD. PMID- 8423874 TI - Wilson's disease: evidence of subgroups derived from clinical findings and brain lesions. AB - Using exploratory factor analysis, we prospectively investigated neuropsychiatric symptoms and structural brain lesions of 47 patients with proven Wilson's disease and identified three subgroups. The first subgroup clinically exhibited bradykinesia, rigidity, cognitive impairment, and an organic mood syndrome and by MRI showed a dilatation of the third ventricle. The second subgroup was characterized by ataxia, tremor, reduced functional capacity, and focal thalamic lesions. The third subgroup showed dyskinesia, dysarthria, an organic personality syndrome, and focal lesions in the putamen and in the pallidum. PMID- 8423875 TI - Central basis of muscle fatigue in chronic fatigue syndrome. AB - We studied whether muscle fatigue, metabolism, or activation are abnormal in the chronic fatigue syndrome (CFS). Subjects performed both an intermittent submaximal and a sustained maximal voluntary isometric exercise protocol of the tibialis anterior muscle. The extent of fatigue, metabolic response, and changes in both M-wave amplitude and twitch tension during exercise were similar in patients and controls. The response to systemic exercise was also normal in the patients. However, voluntary activation of the tibialis was significantly lower in the patients during maximal sustained exercise. The results indicate that patients with CFS have (1) normal fatigability and metabolism at both the intracellular and systemic levels, (2) normal muscle membrane function and excitation-contraction coupling, and (3) an inability to fully activate skeletal muscle during intense, sustained exercise. This failure of activation was well in excess of that found in controls, suggesting an important central component of muscle fatigue in CFS. PMID- 8423876 TI - Education and the prevalence of dementia and Alzheimer's disease. PMID- 8423877 TI - Idiopathic postural orthostatic tachycardia syndrome: an attenuated form of acute pandysautonomia? AB - To characterize the idiopathic postural orthostatic tachycardia syndrome (POTS), we reviewed the records of all patients aged 20 to 51 who presented to the Mayo Autonomic Reflex Laboratory and who exhibited tachycardia at rest or during head up tilt. These patients were usually women who experienced an acute onset of persistent lightheadedness and fatigue or gastrointestinal dysmotility. In seven patients, a viral illness may have preceded the onset of symptoms. In two instances, signs and symptoms of a small-fiber sensory neuropathy were present. Laboratory evaluation of autonomic function revealed increased diastolic blood pressure to tilt (5/16), increased Valsalva ratio, marked decrease in phase II of the Valsalva maneuver with normal phase IV overshoot, and normal forced respiratory sinus arrhythmia. Abnormal quantitative sudomotor axon reflex test and thermoregulatory sweat test and an excessive orthostatic increase of catecholamines were found in some patients. We conclude that in many instances POTS may be a manifestation of a mild form of acute autonomic neuropathy. PMID- 8423878 TI - Functional status, education, and the diagnosis of dementia in the Shanghai survey. AB - We examined the relationship of culturally adapted Chinese versions of the Mini Mental State Examination (CMMS) and several functional measures to the effect of education on the clinical diagnosis of dementia in 554 subjects (55 to 95 years; median, 74) who had undergone intensive evaluation during the Shanghai survey of dementia. Low education was associated with increased prevalence of clinically diagnosed dementia. The standardized history and one functional scale (Pfeffer Outpatient Disability Scale [POD]) clustered closely with clinical diagnosis on factor analysis, whereas the CMMS, Instrumental Activities of Daily Living scale (IADL), and Activities of Daily Living scale (ADL) loaded additionally onto an education-weighted component. A logistic equation based on the CMMS, history, POD, and IADL was the best predictor of the clinical diagnosis of dementia, but history, POD, and IADL without a mental status score also predicted the diagnosis with a sensitivity of 88.6%, a specificity of 89.3%, a positive predictive value of 66.0%, and a negative predictive value of 97.1%. When dementia was diagnosed using an algorithm based on the three functional scales alone, low education continued to be associated with increased age-specific risk of dementia. PMID- 8423879 TI - In vitro MR microscopy of the hippocampus in Alzheimer's disease. AB - We used MR microscopy at 7 tesla to identify the anatomy of the degenerating hippocampus in Alzheimer's disease (AD), which we then correlated with the histopathologic findings in the same specimens. The specimens studied were resected postmortem from 13 patients with confirmed AD and from nine age-matched controls. We imaged the specimens in the coronal plane using either three dimensional Fourier encoding or single-slice Carr, Purcell, Meiboom, Gill (CPMG) spin echo sequences. On all specimens imaged with the CPMG pulse sequence, we calculated the T2 relaxation times for subfields within the hippocampus. Histologic sections were taken from each specimen and compared with the corresponding MR image. Using histologic boundaries, we quantified the number of neuritic plaques and neurofibrillary tangles in each hippocampal subfield. We measured the area, morphometric characteristics, and width of identifiable signal variant regions on each image and compared these measurements with the histopathologic findings. The mean cross-sectional area of the hippocampus in AD was decreased by 31% compared with the control group. This atrophy was highly correlated with tangle counts within the hippocampus, but not with plaque counts. The width of the gray matter in hippocampal area CA1, as identified by MR, correlated with the total area of the hippocampus. An age-related decrease in the size of a low-signal region that corresponds histologically to input projections comprising part of the perforant pathway was identified. Measurements of the T2 relaxation times of hippocampal subfields showed little regional variability and were not accurate indicators of disease presence or severity (p > 0.05). PMID- 8423880 TI - Phosphatidylserine, a putative inhibitor of tumor necrosis factor, prevents autoimmune demyelination. AB - We tested the effect of bovine cortex phosphatidylserine (BC-PS), a membrane phospholipid known to inhibit the release of the cytokine tumor necrosis factor (TNF), in SJL/J mice sensitized for adoptively transferred experimental autoimmune encephalomyelitis (EAE). Control, sensitized mice developed severe clinical and histologic EAE within 6 to 9 days, whereas only 20% of mice given BC PS displayed clinical signs that were much less severe and minimal CNS pathology. Cessation of BC-PS treatment after 15 to 40 days led to disease within 1 to 2 weeks, but when treatment was more prolonged, animals remained healthy after cessation. Serial transfer of spleen cells (SC) from BC-PS-treated and control animals into naive recipients resulted in acute EAE within 6 to 7 days with cells from either donor type. Animals treated late with BC-PS failed to relapse and generally remained healthier than controls did over a 40-day period of observation. Cultures of lymph node cells or SC from BC-PS-treated and control animals showed an 80 to 90% reduction in TNF production in the BC-PS-treated group. Thus, we demonstrate that PS can abrogate or significantly reduce the severity of EAE without permanently inhibiting effector T cells. The approach might be considered a candidate for future therapies of relevance to MS. PMID- 8423881 TI - Borrelia burgdorferi-specific intrathecal antibody production in neuroborreliosis: a follow-up study. AB - We used a capture ELISA with biotinylated Borrelia burgdorferi flagella as antigen to analyze the kinetics of intrathecal antibody production against B burgdorferi in 27 patients with neuroborreliosis. All patients had lymphocytic pleocytosis, 13/27 had intrathecal specific IgM production, and 26/27 had intrathecal IgG synthesis against B burgdorferi before therapy. All patients improved after antibiotic treatment. At follow-up, 11 months to 8 years later (median, 1 1/2 years), 20 patients had had a complete clinical recovery, and seven suffered from sequelae. One patient without sequelae had persistent specific intrathecal IgM synthesis. Ten of 20 patients without sequelae and five of seven patients with sequelae had persistent intrathecal IgG production against B burgdorferi. None of the 16 patients with persistent specific intrathecal antibody synthesis had pleocytosis at follow-up. Therefore, intrathecal immunoglobulin production against B burgdorferi, especially IgG, may persist for years after treatment of neuroborreliosis without clinical signs of active disease. PMID- 8423882 TI - Botulinum toxin in the treatment of writer's cramp: a double-blind study. AB - We treated 20 patients with writer's cramp in a double-blind, placebo-controlled study. Each patient received two treatments in tandem, one with botulinum-A toxin (BTX-A) injections and another with normal saline, separated by 3 months. Treatment order was randomized and unknown to the patient and physician. Patients were assessed before each treatment and 2 and 6 weeks after each treatment by objective measurements of pen control. Twelve patients had improvement in pen control after treatment with BTX-A, but only four had significant improvement in writing. BTX-A injections are effective in relieving symptoms in selected cases of writer's cramp, particularly in those with significant wrist-joint deviation. PMID- 8423883 TI - Cortical hyperexcitability in progressive myoclonus epilepsy: a study with transcranial magnetic stimulation. AB - In progressive myoclonus epilepsy (PME), responses to afferent input are frequently abnormal. It is unclear whether the abnormality lies at the cortical, subcortical, or segmental level. To obtain evidence for an exaggerated effect on motor cortical excitability, we used peripheral nerve and transcranial magnetic stimulation in controls and subjects with idiopathic generalized epilepsy and PME. Mean threshold intensity was higher in those with idiopathic generalized epilepsy and PME than in controls, probably as a result of anticonvulsant treatment. A long-latency response to peripheral stimulation and an exaggerated facilitatory effect of peripheral stimulation on the motor evoked potential was present in subjects with PME. Latency differences between the late responses in the upper and lower limbs provided evidence against a segmental reflex and implicated rapidly conducting fibers in the spinal cord. Both the late response and the facilitatory effect had onset latencies consistent with a transcortical pathway, suggesting an exaggerated effect of afferent input on motor cortical excitability in PME. PMID- 8423884 TI - Quantitative synaptic alterations in the human neocortex during normal aging. AB - We quantified the synaptic population density in the frontal cortex of 25 individuals without dementia 16 to 98 years old, using sections double immunolabeled for beta/A4 amyloid and for synaptophysin, and found a significant inverse correlation between the presynaptic terminal (PT) counts and age (r = 0.7, p < 0.001). Individuals older than 60 years had an average 20% decrease in PT density compared with individuals younger than 60 years. There were no significant correlations between the age and the number of beta/A4 amyloid positive plaques or between synaptic density and the number of amyloid plaques. Further analysis of the digitized serial optical images showed focal areas of synapse loss and distended synaptophysin-containing boutons in the mature plaques of the normal aged cases. However, we found no microscopic changes in the synaptic content inside and outside the diffuse plaques. We suggest that a loss of synaptic input in the neocortex is an age-dependent factor that contributes to the overall synaptic loss in Alzheimer's disease, but that this might be largely independent of the beta/A4-amyloid deposition. PMID- 8423885 TI - Effects of estradiol and progesterone on seizure sensitivity in oophorectomized DBA/2J mice and C57/EL hybrid mice. AB - We investigated the effects of 17 beta-estradiol (E2) and progesterone (PG) on seizure sensitivity in two genetically epilepsy-prone strains, the DBA/2J and the C57/EL hybrid. In the DBA/2J, subject to audiogenic seizures when juvenile, oophorectomy produced a marked decrease in seizure sensitivity, both with and without E2 or PG replacement. In the C57/EL, subject to vestibular seizures, E2 significantly reduced seizure frequency and increased lag time to seizure onset. PG did not affect these variables. Both E2 and PG significantly prolonged seizure duration. These results support a role of ovarian hormones in regulating paroxysmal activity in collicular and tegmental regions associated with audiogenic seizures in the DBA/2J and in temporal structures associated with vestibular seizures in the C57/EL. PMID- 8423886 TI - Creutzfeldt-Jakob disease in a physician: a review of the disorder in health care workers. AB - We describe a 58-year-old physician who developed Creutzfeldt-Jakob disease (CJD) 30 years after formal training in pathology. To the best of our knowledge, he is the sixth physician and the 24th health care worker reported with CJD. We summarize the available data on these health care workers. PMID- 8423887 TI - Antineuronal (anti-Ri) antibodies in a patient with steroid-responsive opsoclonus myoclonus. AB - A 45-year-old woman developed opsoclonus, myoclonus, and severe truncal and gait ataxia. Serum and CSF contained IgG antibodies that appear to be identical to "anti-Ri" antibodies associated with paraneoplastic opsoclonus and ataxia. The patient had a fluctuating course with exacerbations that responded well to corticosteroids and later to cyclophosphamide. Her anti-Ri antibody titer has declined significantly but still remains high. After more than 3 years of follow up, no neoplasm has been detected. PMID- 8423888 TI - Early combination therapy (bromocriptine and levodopa) does not prevent motor fluctuations in Parkinson's disease. AB - Early combination therapy with bromocriptine (Br) and levodopa (LD) is believed to delay or prevent the onset of late treatment complications typically associated with LD monotherapy in Parkinson's disease (PD). Studies recommending this regimen have been uncontrolled. We evaluated this possibility in a 4-year, double-blind, randomized, parallel group trial comparing Br and LD both alone and in combination in 22 PD patients never before treated with dopaminergic medications. In the group receiving Br monotherapy, 17% had motor fluctuations (end-of-dose failure or on-off), 17% chorea, 33% dystonia, and 83% freezing. In the LD group, 33% had motor fluctuations, 56% chorea, 100% dystonia, and 22% freezing. In the combination group, 71% had motor fluctuations, 57% chorea, 71% dystonia, and 57% freezing. The frequency of dystonia was significantly lower with Br monotherapy than in the other two treatment groups. No other significant differences were observed. LD monotherapy appeared to have superior efficacy in the treatment of PD. Mean final doses of LD and Br were similar for the different treatment groups. Early combination therapy does not prevent or delay the onset of motor fluctuations or dyskinesia in PD. PMID- 8423889 TI - Increase of Parkinson disability after fluoxetine medication. AB - Depression is a major clinical feature of Parkinson's disease. We report the increased amount of motor disability in four patients with idiopathic Parkinson's disease after exposure to the antidepressant fluoxetine. The possibility of a clinically relevant dopamine-antagonistic capacity of fluoxetine in Parkinson's disease patients must be considered. PMID- 8423890 TI - Spared awareness for the left side of internal visual images in patients with left-sided extrapersonal neglect. AB - Two patients with neglect for extrapersonal space following right-hemisphere stroke both demonstrated retained ability to attend to the left side of internally generated visual images. The attentional system for extrapersonal space may not be identical to the attentional system for intrapersonal space. PMID- 8423891 TI - Intermittent downbeat nystagmus due to vertebral artery compression. AB - Downbeat nystagmus (DBN) uncommonly occurs as a transient phenomenon, and it rarely occurs in patients with cerebrovascular disease. We observed a patient with intermittent DBN and lightheadedness due to transient obstruction of his dominant vertebral artery when he turned his head to his left side. Surgical removal of an osteophyte at the site of the angiographically demonstrated lesion relieved his symptoms. PMID- 8423892 TI - Kearns-Sayre syndrome with a phenocopy of choroideremia instead of pigmentary retinopathy. AB - Mitochondrial DNA (mtDNA) was deleted in a patient with Kearns-Sayre syndrome (KSS) presenting with a choroideremia-like fundus picture instead of pigmentary retinopathy. No evidence for X-linked choroideremia was present, and because of the strong association between KSS and deleted mtDNA, we suggest that choroideremia is a phenocopy and can be part of KSS. PMID- 8423893 TI - Position statement: certain aspects of the care and management of profoundly and irreversibly paralyzed patients with retained consciousness and cognition. Report of the Ethics and Humanities Subcommittee of the American Academy of Neurology. PMID- 8423894 TI - Competent patients with advanced states of permanent paralysis have the right to forgo life-sustaining therapy. PMID- 8423895 TI - Brodmann's "missing" numbers. PMID- 8423896 TI - When selegiline is prescribed, Stelazine may be mistakenly dispensed. PMID- 8423897 TI - Transient monocular blindness associated with cocaine abuse. PMID- 8423898 TI - Stimulus-specific pathologic laughter: a case report with discrete unilateral localization. PMID- 8423899 TI - Oral megadose methylprednisolone therapy in a patient with chronic progressive multiple sclerosis. PMID- 8423900 TI - Lacunes. PMID- 8423901 TI - TPA in stroke. PMID- 8423902 TI - Platelet membrane fluidity. PMID- 8423903 TI - Platelet membrane fluidity. PMID- 8423904 TI - Nerve agents. PMID- 8423905 TI - Tourette's and stress. PMID- 8423906 TI - The efficacy and safety of ticlopidine and aspirin in non-whites: analysis of a patient subgroup from the Ticlopidine Aspirin Stroke Study. AB - We analyzed the efficacy of ticlopidine and aspirin in the non-white subgroup of patients from the Ticlopidine Aspirin Stroke Study. In this double-blind, randomized, multicenter study, patients received either ticlopidine 250 mg (312 non-white patients) or aspirin 650 mg (291 non-white patients) twice a day. The 1 year cumulative event rate per 100 patients for nonfatal stroke or death from any cause was 5.5 for ticlopidine and 10.6 for aspirin--an apparent 48.1% reduction in risk with ticlopidine relative to aspirin. The 1-year cumulative event rate for fatal or non-fatal stroke was 3.7 for ticlopidine and 9.4 for aspirin--an apparent 60.8% reduction in risk with ticlopidine relative to aspirin. The cumulative event rates for both endpoints also were lower in ticlopidine-treated patients after the 2nd and 3rd years. These reductions were not significantly different between treatment groups, but were of the same order of magnitude as previously found for the total series, which did attain statistical significance (p = 0.048), and the frequency of adverse events was not significantly different between the two treatment groups. Severe neutropenia, the most serious adverse event associated with ticlopidine use, did not occur in non-white patients. These results suggest that ticlopidine is superior to aspirin for stroke prevention in non-whites. PMID- 8423907 TI - Ischemic stroke in patients with atrial fibrillation: effect of aspirin according to stroke mechanism. Stroke Prevention in Atrial Fibrillation Investigators. AB - Ischemic strokes occurring in patients with nonrheumatic atrial fibrillation are due to a variety of mechanisms, not exclusively to cardiogenic embolism. Without knowledge of antithrombotic therapy assignment, we categorized strokes in the Stroke Prevention in Atrial Fibrillation Study as presumed cardioembolic or noncardioembolic. We then compared patient clinical and echocardiographic variables, as well as the efficacy of aspirin prophylaxis, for each stroke type. Of 71 ischemic strokes, we categorized 46 (65%) as cardioembolic, 13 (18%) as noncardioembolic, and 12 (17%) as of uncertain cause. Patients developing noncardioembolic strokes, relative to cardioembolic strokes, were more commonly men (p = 0.005) and were more likely to have left ventricular wall motion abnormalities by two-dimensional echocardiography (p = 0.002). Aspirin reduced the occurrence of strokes categorized as noncardioembolic significantly more than it did those categorized as cardioembolic (p = 0.01). These results emphasize the value of considering stroke mechanisms in therapeutic trials of antithrombotic agents and suggest a differential effect of aspirin according to mechanism. PMID- 8423908 TI - Alternating hemiplegia of childhood: a study of 10 patients and results of flunarizine treatment. AB - Alternating hemiplegia of childhood is a rare syndrome characterized by onset before 18 months of age of frequent attacks of alternating paralysis, transient ocular palsies, nystagmus, choreoathetosis, and autonomic dysfunction. We describe features of 10 patients followed for up to 27 years. The mechanism of alternating hemiplegia remains unknown but an association to migraine is suspected because of the strong family history of migraine and aura symptoms in some patients. We treated nine patients with flunarizine, a calcium channel blocker, for up to 5 years; they showed a reduction in duration of the hemiplegic attacks, but the episodes ceased completely in only one patient. With long-term follow-up, the persistent motor, movement, and cognitive deficits are more apparent. It is not certain if the flunarizine alters this course. PMID- 8423909 TI - Withdrawal of antiepileptic medication in children--effects on cognitive function: The Multicenter Holmfrid Study. AB - We present 100 children diagnosed with epilepsy who were seizure-free for more than 1 year and still on monotherapy of antiepileptic drugs (AEDs). We matched each child with a healthy classmate and performed neuropsychological testing and EEG before and after complete withdrawal of the AEDs. The withdrawal phase lasted 3 months, but the dose decrease was individualized for each patient. Three to 4 months after complete withdrawal of the drug all patients were reassessed. Patients with seizure relapse are excluded from the study. Seventeen patients are regarded as dropout, 11 because of seizure relapse and six because of protocol violation. The remaining 83 patients were treated with carbamazepine (n = 56), valproic acid (n = 17), or phenytoin (n = 10). Serum concentrations of the AEDs were measured using peak plasma levels that were taken immediately before or after psychological testing. We used neuropsychological tests to assess psychomotor function and "central" cognitive processing such as information processing or memory function. We found significant improvement attributable to drug withdrawal on only one of the cognitive tests, namely, psychomotor speed, suggesting that the impact of AED treatment on higher-order cognitive function is rather limited. In addition, we found group differences between the epilepsy group and the control group at baseline that persisted after drug withdrawal. Subsequent analysis showed some factors that may have contributed to these group differences. First, patients with a former diagnosis of absence seizures show lower scores both at baseline and after drug withdrawal. We may assume that the seizure propensity has not disappeared completely in these patients. Some evidence is found that phenytoin may have a different cognitive profile than carbamazepine, with more impairment on tests that measure motor and mental speed. Again, this impairment persists after drug withdrawal. PMID- 8423910 TI - Cheiro-oral topography of sensory disturbances due to lesions of thalamocortical projections. AB - Sensory disturbance in the unilateral hand and ipsilateral mouth region, the cheiro-oral syndrome, may be due to cortical, thalamic, or brainstem lesions. We report five patients with this syndrome due to infarction at the border of the posterior limb of the internal capsule and the corona radiata. The sensory fibers from the mouth area and hand probably travel to the cortical sensory areas in close proximity after leaving the posterior ventral thalamic nucleus. PMID- 8423911 TI - The positive diagnosis of narcolepsy and narcolepsy's borderland. AB - We studied the validity of cataplexy and number of sleep-onset rapid-eye-movement periods (SOREMPs) during one Multiple Sleep Latency Test (MSLT) as determinants of narcolepsy in 306 subjects with excessive daytime sleepiness not related to obstructive sleep apnea or other known syndromes. The subgroup defined by a history of cataplexy was the most homogeneous in clinical and polygraphic variables. However, only 83% of these subjects had two or more SOREMPs in one MSLT. The subgroup defined by two or more SOREMPs included many patients without cataplexy. A disproportionate number of these subjects were older women whose chances of developing cataplexy are remote. This group of older women had a higher number of periodic leg movements during sleep than the other groups. Patients with both cataplexy and two or more SOREMPs have the greatest chance of being DR2 DQw1 positive. Thus, the combination of history of cataplexy and two or more SOREMPs is the best clinical determinant of narcolepsy. However, two or more SOREMPs is a poorer discriminant of narcolepsy than history of cataplexy. PMID- 8423912 TI - The effect of cholesterol-lowering agents on hepatic and plasma cholesterol in Niemann-Pick disease type C. AB - Niemann-Pick disease type C (NP-C) is a neurovisceral lipidosis characterized by defective intracellular trafficking of cholesterol and lysosomal accumulation of unesterified cholesterol, believed to be an offending metabolite. We studied the effect of cholesterol-lowering agents on hepatic and plasma cholesterol levels in NP-C by randomly assigning 25 patients with NP-C to one of five treatment regimens containing different combinations of cholestyramine, lovastatin, nicotinic acid, or dimethyl sulfoxide (DMSO). Unesterified cholesterol content was measured in liver biopsies before and after 4 months' treatment. All drug regimens except DMSO alone reduced hepatic and plasma cholesterol levels. Toxicity was limited and did not prevent any patient from completing the study. The combination of cholestyramine, lovastatin, and nicotinic acid lowered cholesterol levels in liver and blood with minimal side effects. A controlled clinical study will be necessary to determine if this regimen influences the rate of neurologic progression. PMID- 8423913 TI - Hyperintense globus pallidus on T1-weighted MRI in cirrhotic patients is associated with severity of liver failure. AB - Hyperintense globus pallidus on T1-weighted MRI is present in most patients with advanced liver disease. We evaluated the relationship between the signal intensity of the globus pallidus and clinical or laboratory data of 77 patients eligible for liver transplantation. There was a significant correlation between the intensity of the signal and the Child-Pugh score (as indication of severity of liver disease), presence of postural tremor, previous episodes of variceal bleeding or hepatic encephalopathy, prothrombin activity, serum aspartate and alanine aminotransferase, bilirubin, and the indocyanine green (ICG) hepatic clearance, a very sensitive marker of liver function. The multivariate analysis disclosed that the ICG hepatic clearance and previous episodes of variceal bleeding were independently associated with the signal intensity in the globus pallidus. MRI repeated in 21 patients 10 to 20 months after transplant showed a disappearance of the lesion in all cases. We conclude that the hyperintense globus pallidus is secondary to the severity of the liver disease, and is reversible when liver function returns to normal. PMID- 8423914 TI - Dominantly inherited, early-onset parkinsonism: neuropathology of a new form. AB - We report postmortem findings in a 46-year-old man with dominantly inherited parkinsonism whose symptoms started at age 28. At least 13 other family members in three generations have been affected, some from early childhood. Dystonia is a prominent feature in several of the youngest patients, but was not present in this patient. After several years of successful treatment with medication, he developed severe on-off fluctuations and dyskinesias. At age 45, the patient underwent stereotaxic implantation of autologous adrenal medullary tissue into the left corpus striatum and lateral ventricle. He improved considerably over the following 6 months, but then developed glioblastoma multiforme and died 1 year after transplantation. There was severe neuronal loss in the pars compacta and pars reticulata of the substantia nigra, with prominent gliosis in the pars reticulata. The nigral neurons remaining in the pars compacta were poorly pigmented. Neither Lewy bodies nor neurofibrillary tangles were present, and we identified no other degenerative neuropathologic changes. This combination of pathologic and clinical features differs from any previously reported case. PMID- 8423915 TI - Mesiobasal versus lateral temporal lobe epilepsy: metabolic differences in the temporal lobe shown by interictal 18F-FDG positron emission tomography. AB - Metabolic abnormalities in the temporal lobe (TL) of 25 patients suffering from temporal lobe epilepsy of mesiobasal or lateral TL origin have been investigated using interictal [18F]-fluorodeoxyglucose (FDG) positron emission tomography (PET). The epileptogenic area was determined by ictal EEG recordings using foramen ovale and scalp electrodes in 20 patients, and by the use of stereo electroencephalography in one patient. Four patients with structural lesions on their MRIs had noninvasive ictal surface EEG recordings. Sixteen patients had a clear-cut mesiobasal seizure onset, and in five patients the seizures originated from the lateral temporal neocortex. Twenty-four patients underwent selective surgery. Patients with temporal limbic seizures associated with mesial gliosis (n = 15) had the lowest FDG uptake in the entire TL, followed by patients with lateral temporal seizure origin (n = 5). Patients with tumors located in the mesiobasal TL (n = 5) showed, in general, only a slight decrease of glucose metabolism in all TL structures. There was no clear-cut correlation between the degree of hypometabolism and the location of EEG-defined epileptogenic focus. The metabolic pattern, however, differed between the patient groups and allowed a discrimination between patients of mesial temporal and lateral temporal seizure onset. PMID- 8423916 TI - Quiescent and cycling cell compartments in the senescent and Alzheimer-diseased human brain. AB - Statin is a 57-kd nuclear protein expressed exclusively in nonproliferating cells. In the present study, immunohistochemical localization of statin in normal, senescent human brain revealed that virtually all neurons, ependymal cells, vascular smooth muscle cells, and endothelial cells are statin-positive and, hence, postmitotic. As we previously demonstrated in rodents, an unexpectedly large fraction of neuroglial cells throughout the aging human brain is statin-negative (range, 41 to 45%), consistent with the substantial retention of neuroglial proliferative capacity well into the senium. In Alzheimer's disease, there is a significant increase in the proportion of statin-negative neuroglia (range, 51 to 58%). In all regions except cerebellum, loss of statin in Alzheimer neuroglia could be accounted for by changes involving the astrocyte subpopulation. These results provide evidence that reactive gliosis in this neurodegenerative disorder encompasses some degree of astrocyte hyperplasia in addition to astrocyte hypertrophy. Maintenance of normal compartments of cycling and quiescent neuroglia in the senescent human brain may serve to define neurologic well-being during the aging process. Conversely, deviations in neuroglial cytokinetics may indicate the presence and extent of intervening neuropathologic processes. PMID- 8423917 TI - Residual impairments and work status 15 years after penetrating head injury: report from the Vietnam Head Injury Study. AB - We investigated the relationship of neurologic, neuropsychological, and social interaction impairments to the work status of a large sample of penetrating head injured patients wounded some 15 years earlier during combat in Vietnam. Extensive standardized testing of neurologic, neuropsychological, and social functioning was done at follow-up on each head-injured patient (N = 520), as well as on a sample of uninjured controls (N = 85). Fifty-six percent of the head injured patients were working at follow-up compared with 82% of the uninjured controls. Seven systematically defined impairments proved to be most correlated with work status. These were post-traumatic epilepsy, paresis, visual field loss, verbal memory loss, visual memory loss, psychological problems, and violent behavior. These disabilities had a cumulative and nearly equipotent effect upon the likelihood of work. We suggest that a simple summed score of the number of these seven disabilities can yield a residual "disability score" which may prove to be a practical tool for assessing the likelihood of return to work for patients in this population and perhaps in other brain-injured populations. These findings may also help to focus rehabilitation efforts on those disabilities most likely to affect return to work. PMID- 8423918 TI - Suicide in custody: occurrence in Maori and nonMaori New Zealanders. AB - AIMS: To assess whether Maori people are particularly vulnerable to suicide in custody. METHODS: Suicides in New Zealand prisons (1973-88) and police cells (1980-91) were analysed in order to compare the risks for Maori and nonMaori men. RESULTS: Among prisoners, the risk of suicide for Maori men was very similar to that for nonMaori (38.7 Maori and 40.4 nonMaori suicides per 100,000 male distinct prisoners received per year). Maori men were, however, six times more likely than nonMaori to be in prison, with half of all sentenced prisoners being Maori. This explains why the proportion of the entire Maori population who died as a result of prison suicide was eight times higher than that for the nonMaori population. For police cells it was not possible to calculate suicide rates except as proportions of the entire Maori and nonMaori populations. The proportion of Maori men who died in police custody was nine times higher. Suicide in custody accounted for about a quarter of all suicides in Maori men aged 15-49 years, but only 1.7% of suicides in nonMaori men. CONCLUSION: These results suggest that reduction of Maori suicides in custody is most likely to be achieved by tackling the overrepresentation of Maori people in custody. PMID- 8423919 TI - A team process for the introduction of the laparoscopic cholecystectomy technique to a public hospital service. AB - AIMS: To introduce the technique of laparoscopic cholecystectomy to Christchurch hospitals by a process involving all surgeons in the general surgical team. To assess prospectively the potential for cost-saving. METHOD: A visiting surgeon with experience of the procedure, demonstrated the technique and supervised operations by members of the team on twenty six informed consenting patients. RESULTS: The process has enabled all general surgeons to gain competence in the technique, with low complication rates. CONCLUSIONS: The process adopted in Christchurch quickly familiarised all general surgeons with the technique of laparoscopic cholecystectomy, so that it was safely and efficiently made available to all patients who could benefit. The capacity of the procedure to reduce the cost per case treated was assessed, and reduced social cost to patients demonstrated. PMID- 8423921 TI - Superficial dermatophyte infections. PMID- 8423920 TI - Antibiotic protocol for appendicectomy--costs and benefits of single dose therapy. AB - AIMS: To examine possible savings from the introduction of an antibiotic protocol for one surgical procedure and to define antibiotic charting techniques. METHODS: An antibiotic policy for patients undergoing an appendicectomy was instituted in the surgical unit at North Shore Hospital to examine possible savings and to define antibiotic charting techniques. Depending on the macroscopic findings at surgery a third generation cephalosporin was used either as a single dose intraoperatively or for multiple doses postoperatively. RESULTS: The case notes of a control group of 55 patients were reviewed. This showed an average antibiotic cost per appendicectomy of $69.00 ($52.00) for nongangrenous nonperforated appendices (NGNPA). During the study period 57 patients underwent appendicectomy at an average antibiotic cost of $51.00 ($32.00 for NGNPA). This represented a 26% saving per patient during the study period and a 39% saving for those with NGNPA: CONCLUSIONS: The findings advocate the use of frequently updated guidelines for antibiotic use in surgical procedures. It reaffirms that medical staff must chart precisely as poor charting represents significant waste of expenditure in hospitals. PMID- 8423922 TI - Health reforms and the public hospitals. PMID- 8423923 TI - Health reforms. PMID- 8423924 TI - Asthma in the Maori. PMID- 8423925 TI - Control of epidemic group A meningococcal disease in Auckland. AB - AIM: To study group A meningococcal vaccine delivery to infants less than 2 years of age in Auckland in 1987 to control epidemic disease. METHODS: Mechanisms of vaccine delivery and its facilitation are described. A detailed audit of delivery of vaccine to children less than two years using signed consent forms determined delivery source. This was the age group at highest risk, and poorly covered by routine childhood vaccines. Primary health care source of children presenting with disease was determined by telephone. RESULTS: The epidemic of group A meningococcal disease in the winters of 1985 and 1986 abated most likely due to the vaccination of high risk children (3 months-13 years) in 1987. 90% of the target population were vaccinated. In south Auckland the majority (92%) of vaccine doses for children less than two years of age was delivered by the Plunket Society with Department of Health backing aided by community health workers. By contrast delivery by, general practitioners was greater in north-west and central Auckland (approximately 25%, of dose 1), especially after the publicity over possible side effects (approximately 50% of dose 2). Coverage for dose 1 of children < 2 years was similar (89%) in south Auckland. Of children presenting with meningococcal disease 1 in 4 did not have an identifiable general practitioner. CONCLUSIONS: Vaccines to prevent serious paediatric illness are known to be highly cost effective. The best method of delivery of vaccinations may vary from area to area. Major community involvement including community health workers for the Maori and Pacific Island communities may have facilitated the dissemination of information in this campaign. PMID- 8423926 TI - Evaluation of the cot death prevention programme in South Auckland. AB - AIMS: The National Cot Death Prevention Programme aims to reduce the prevalence of four modifiable risk factors for cot death, namely infant sleeping prone, maternal smoking, lack of breast feeding and infant sharing a bed with another person. This study evaluated the knowledge of 200 mothers of infants in South Auckland and estimated the prevalence of these infant care practices, which were compared with that found in the New Zealand Cot Death Study. METHODS: 200 mothers were interviewed. RESULTS: The prevalence of these modifiable risk factors in this study and that found in 1987/89 in Auckland were: prone sleep position: 2.5% compared with 36.8%, p < 0.001; infant sharing a bed with another person: 23.5% and 45.2%, p < 0.001; maternal smoking: 24.0% and 26.1%, p = ns; lack of breast feeding at 4 weeks of age: 11.0% and 13.6%, p = ns. The following percentage of mothers knew that there were risk factors for cot death: prone sleep position 95.5%, maternal smoking 89.4%, lack of breast feeding 63.1% and infant sharing a bed with another person 68.0%. CONCLUSIONS: This study shows that infant care practices are changing and highlights the need for continuing efforts, especially relating to maternal smoking and the practice of infants sharing a bed with another person. PMID- 8423927 TI - Reporting of lung scans: the future beckons. PMID- 8423928 TI - The value of thallium scintigraphy in a district general hospital. AB - The usefulness of exercise Tl scintigraphy (combined with electrocardiography) in a district general hospital has been assessed in 80 patients in whom a previous exercise electrocardiogram (ECG) had not been definitive. Seventy-five of the patients presented with chest pain and five with no pain but an abnormal ECG. Case notes and test results were examined to establish the clinicians' judgment of the likelihood of having coronary heart disease (CHD) and the patient's outcome. The mean time of follow-up was 27 months. In the majority of patients the test was found to help the clinician come to a decision on the presence or absence of CHD. It was found to be particularly useful in patients with abnormal resting ECGs. There was a shorter time to discharge for patients classified as highly unlikely of having CHD. Only four patients were re-referred following discharge from the outpatient clinic. Nine patients were referred for coronary angiography with five being found to have significant stenosis. None of the 16 patients in whom the exercise ECG was normal had ischaemic myocardial segments in the Tl scintigraphy. PMID- 8423929 TI - Pulmonary uptake of sestamibi on early post-stress images: angiographic relationships, incidence and kinetics. AB - Early post-stress imaging with 99Tcm-sestamibi has the potential to reveal ancillary markers of severe coronary artery disease. Lung/myocardial ratios of sestamibi were assessed after pharmacologic, exercise or combined stress, and these were compared with historical controls who were stressed similarly, but imaged with 201Tl. Forty initial patients had planar imaging and correlating angiograms; pulmonary uptake for sestamibi related to severe coronary artery stenoses when measured on immediate images, started at 4 min post-stress (P = 0.04), but had a poor relationship to angiographic findings when measured on delayed clinical images. Of 180 subsequent studies, increased pulmonary uptake of sestamibi was seen more frequently (incidence = 34%) in those with abnormal tomograms compared to those with normal tomograms (incidence = 13%, P < 0.01), but appeared less frequently than on abnormal 201Tl studies (incidence = 60%). With sequential imaging for 5 min after injection, pulmonary uptake showed a greater fall with time on sestamibi studies than on matched 201Tl studies. No consistent differences were seen among the stress modalities. As an ancillary sign of haemodynamically severe disease, increased pulmonary uptake can be seen after various stress modalities, but may be more difficult to apply with sestamibi than with thallium imaging. PMID- 8423930 TI - Correlation of 99Tcm-sestamibi SPECT with coronary angiography in general hospital practice. AB - One hundred and twenty-eight patients had a stress test with sestamibi injection and a coronarography within 2 months of the sestamibi study. Sestamibi studies were reconstructed using Hanning 0.65 and Wiener 1.0 filters; the latter improved the results significantly (P < 0.05). Significant correlations were found between sestamibi and coronarography: the sensitivity was 0.967 and the specificity was 0.711. Thus sestamibi is a very good compound, with excellent imaging quality, which is very sensitive for mild defects, even more sensitive than coronarography. Thorough studies of discrepancies show that they could be explained by reductions in coronary flow or myocardial reserve, even when the large epicardial vessels are disease free as occurs in various coronary involvements. A follow-up of positive patients with sestamibi and negative coronarography is absolutely necessary since this group of patients represents a 'risk' group. Moreover, sestamibi single photon emission computed tomographic accuracy is highly dependent on filter choice. PMID- 8423931 TI - 99Tcm-methoxyisobutylisonitrile stress/rest SPECT in patients with constant complete left bundle branch block. AB - Numerous studies have revealed frequent false positive septal findings of 201Tl stress imaging in patients with left bundle branch block (LBBB) even with angiographically excluded significant coronary artery disease (CAD). To scrutinize this phenomenon for stress/rest 99Tcm-methoxyisobutylisonitrile (MIBI) single photon emission computed tomography (SPECT) was used to review 22 patients with constant LBBB. The findings were reversible septal defect in one patient, partially reversible septal defect in one patient and irreversible septal defects in eight patients. In four patients 99Tcm-MIBI scans were entirely normal. Thus, 12/22 (55%) patients revealed normal septal 99Tcm-MIBI uptake. Both patients with a stress-induced septal defect revealed a significant left anterior descending artery stenosis on coronary angiography. These preliminary results suggest, that 99Tcm-MIBI might be more specific and accurate than 201Tl in the evaluation of CAD in patients with LBBB because of apparently rare or absent false positive septal findings. PMID- 8423932 TI - Right ventricular function at rest and during submaximal exercise assessed by 81Krm equilibrium ventriculography in normal subjects. AB - 81Krm equilibrium ventriculography was used to study right ventricular function in 37 healthy male volunteers. 'Anatomical' lung subtraction using 99Tcm lung perfusion scintigraphy was compared with conventional background correction in the calculation of resting right ventricular ejection fraction (RVEF). Resting RVEF was significantly greater following 'anatomical' lung subtraction than that using background correction (0.59 +/- 0.06 versus 0.55 +/- 0.05). The exercise response of the normal right ventricle was defined in 23 subjects during exercise. Right ventricular ejection fraction showed a progressive increase during graded submaximal exercise (0.55 +/- 0.05 at rest, 0.60 +/- 0.05 at 50 W and 0.66 +/- 0.05 at 100 W). Right heart 81Krm equilibrium ventriculography is well suited to the evaluation of right ventricular function at rest and during exercise. The absolute value of RVEF will however be dependent upon the method of image analysis. PMID- 8423933 TI - Unresolved pulmonary embolism: the role of fibrinolysis. AB - A minority of patients with acute pulmonary embolism (PE) show failure of resolution when assessed by serial ventilation/perfusion (V/Q) radionuclide lung imaging. The fibrinolytic systems were studied in six such patients (group I), and in 11 patients in whom PE had resolved (group II), together with 17 healthy control subjects. Assays of the fibrinolytic system included euglobulin clot lysis times (ECLT), tissue plasminogen activator (tPA), and plasminogen activator inhibitor-1 (PAI-1). Euglobulin clot lysis times were not prolonged in the unresolved PE group, but were significantly longer in patients in group II when compared to control subjects (P < 0.03). This could not be explained either on the basis of tPA levels, which were higher in group II when compared to group I (P < 0.05) and control subjects (P < 0.02), or on the basis of PAI-1 levels which did not differ significantly between the three groups. Our inability to demonstrate derangements of fibrinolysis in the patients with unresolved PE makes defective fibrinolysis an unlikely aetiological factor in the persistence of thrombosis in these patients. PMID- 8423934 TI - Detection of nasopharyngeal carcinoma using 99Tcm-methoxyisobutylisonitrile SPECT. AB - 99Tcm-methoxyisobutylisonitrile (MIBI) single photon emission computed tomography (SPECT) of the head and neck was performed in 20 patients with nasopharyngeal carcinoma (NPC) to detect the malignancies. Ten volunteers also underwent 99Tcm MIBI SPECT of the head and neck for comparison. The results showed no MIBI uptake in the nasopharynx of any of the 10 normal volunteers. In the patients with NPC, 6/20 (30%) of the cases had no MIBI uptake in the nasopharynx and 14/20 (70%) had increased MIBI uptake in the nasopharynx. In addition, three cases with cervical metastases were detected by 99Tcm-MIBI. However, MIBI uptake in the NPC was not related to the size or differentiation of the NPC. PMID- 8423935 TI - Regional cerebral blood flow of Alzheimer's disease-like pattern in young patients with Down's syndrome detected by 99Tcm-HMPAO brain SPECT. AB - Fourteen young patients with Down's syndrome (nine males, five females, age range 8-30 years) were studied. 99Tcm-hexamethylpropylene amine oxime (HMPAO) brain single photon emission computed tomography (SPECT) was performed to evaluate regional cerebral blood flow (rCBF). Two experienced nuclear medicine physicians analysed the results of the SPECT by visual interpretation. The SPECT in all 14 patients with Down's syndrome invariably showed significantly unilateral perfusion defects in the temporal-parietal-occipital region, occasionally combined with small perfusion defects over other discrete cerebral areas. In this preliminary study, 99Tcm-HMPAO brain SPECT in the patients with Down's syndrome were abnormal. Patterns of rCBF in Down's syndrome were similar to the usual images of rCBF in Alzheimer's disease reported in the literature. Our findings provide new evidence of the similarities between Down's syndrome and Alzheimer's disease. PMID- 8423936 TI - Pharmacokinetics of the FO23C5 anti-CEA antibody fragment labelled with 99Tcm and 111In: a comparison in patients. AB - The FO23C5 anti-carcinoembryonic antigen (CEA) F(ab')2 antibody was radiolabelled with 111In via diethylenetriaminepentaacetic acid (DTPA) and directly with 99Tcm by stannous ion and mercaptoethanol antibody reduction to compare the pharmacokinetics of these three agents. Four patients received 15 mCi 99Tcm-Fab' 1 week before receiving 1 mCi 111In-F(ab')2. Five additional patients received only the 99Tcm-Fab'. Radiochromatograms by high-performance liquid chromatographic (HPLC) analysis of serum and urine samples from patients receiving 111In were typical of those observed by us previously in connection with other antibodies. The identical analyses of samples from patients receiving 99Tcm showed no differences with the method of reduction and more complex radiochromatograms. In addition to peaks due to a mixture of labelled F(ab')2 and Fab' fragments and, occasionally, immune complexes, there were several peaks due to labelled cysteine and other small labelled species present in both serum and urine. The biodistribution of 99Tcm was as expected for a labelled Fab' fragment: relative to 111In, 99Tcm cleared rapidly from circulation and into kidneys and urine. Liver levels of 111In and 99Tcm were surprisingly similar at 1 day (12 versus 9% ID) although initial 111In levels were lower and increased while 99Tcm levels were higher and decreased. Spleen levels were also similar. In 4/9 patients receiving 99Tcm, hepatobiliary clearance was observed at levels which could confuse interpretation whereas this mode of clearance was observed in only 1/4 patients receiving 111In. Image quality was superior with 111In versus 99Tcm at 1 day postadministration as judged by counting rates and background activity whereas the opposite was true at 2-3 h postadministration. PMID- 8423938 TI - Variables influencing cell binding assays for antibody binding kinetics. AB - Owing to growing interest in the use of cell binding assays for quality testing of radiolabelled antibodies in radioimmunoscintigraphy, factors in the assay which might affect final outcome were evaluated in order to accumulate information for effective and reliable assay design. For unmodified antibody, magnitude of binding at certain antibody concentrations changed with alteration in number of target cells attached to the microtitre plate. The level of signal detection could either be enhanced or suppressed by changing the dilution of the enzyme-linked tester antibody as in the case of the enzyme-linked immunosorbent assay (ELISA). Accordingly, changes in either assay factor resulted in a shift in the slope of the binding curve even when the immunoreactivity of the antibody remained unchanged. A conventional technique of single cell preparation by trypsin-EDTA was investigated for the trypsin action on tumour cell antigenicity. No adverse effects could be observed if dissociated cells were incubated for 24 hours before the assay. Variation in number of target cells seemed to be a key factor in determining the power of the assay to reveal changes in the quality of the radiolabelled antibody. Our findings suggested the need to standardize factors such as the size of the seed, determination of plating efficiency and the time for culture in order to assure interassay reproducibility. PMID- 8423937 TI - Evaluation of 99Tcm nonspecific polyclonal IgG in the detection of rejection in a single lung transplant canine model. AB - Acute rejection is an important cause of graft failure in single lung transplantation, however, current noninvasive tests are neither sensitive nor specific for this diagnosis. The aim of this study was to determine whether 99Tcm labelled human nonspecific polyclonal IgG (99Tcm-IgG) may serve as a marker for acute pulmonary rejection following allotransplantation in a dog model. Seventeen mongrel dogs were studied, including four controls and thirteen dogs which underwent surgery [right autotransplant recipient (n = 4), right unmodified allotransplant recipient (n = 5), and right immunosuppressed allotransplant recipient (n = 4)]. At 6 days following surgery, all dogs received 67Ga-citrate and 99Tcm-IgG. Two days later all dogs were sacrificed. Post-mortem examination revealed acute lung rejection in nine animals. No significant difference was found in the percentage uptake of both 99Tcm-IgG and 67Ga-citrate per gram of tissue between rejecting and nonrejecting transplanted lungs. In cases of moderate to severe rejection (n = 5), only 67Ga-citrate showed a significant difference (P = 0.03) in uptake between rejecting and contralateral native lungs, respectively. We conclude that 99Tcm-IgG does not accurately identify acute lung rejection in the early postoperative period. PMID- 8423939 TI - Disposition of nebulized pentamidine measured using the direct radiolabel 123I iodopentamidine. AB - The pulmonary deposition of nebulized pentamidine (300 mg, Respirgard II nebulizer) was measured in seven human immunodeficiency virus (HIV)-positive men using a new radiopharmaceutical, 123I-iodopentamidine. Mean total pulmonary deposition was 15.3 mg or 5.1% of the initial nebulizer dose. Further studies in two of the patients showed that at 24 h, 87% of deposited 123I was retained in the lungs. Small amounts of activity (expressed as a percentage of the initial nebulizer activity) were also detected over the thyroid (0.4%), bladder (1%) and gut (0.7%). The ratio of 123I activity to pentamidine concentration was similar in the nebulizer solution and urine. These results suggest that 123I-pentamidine may be sufficiently stable in vivo to be used to study the biodistribution of inhaled and parenteral pentamidine in humans. PMID- 8423940 TI - Anthropometric assessment of body size differences of full-term male and female infants. AB - OBJECTIVE: To determine gender-specific differences in anthropometric characteristics of full-term male and female infants. METHODS: Twelve hundred five term newborn infants were examined. All measures of length and skinfold thickness were performed in a standardized manner. RESULTS: After adjusting for confounding variables by regression analysis, we found that nearly all length and circumference measurements were significantly smaller in female infants than in male infants but that subcutaneous fat deposition in female infants was significantly increased. However, there was no difference in the ponderal index between male and female newborns, indicating that this measure does not correlate with newborn fat deposition across the sexes. CONCLUSIONS: Despite being shorter and having smaller circumferences, female infants have more subcutaneous fat than male infants. The ponderal index is not useful as a measure of fatness when the sexes are compared. We speculate that the greater subcutaneous fat deposition in female infants may be related to their better neonatal outcomes. PMID- 8423941 TI - Indicators of maternal nutritional status and birth weight in term deliveries. AB - OBJECTIVE: To determine the relationship between measures of maternal protein nutriture and fetal size at birth. METHODS: We obtained serum samples at 18 and 30 weeks' gestation from 289 indigent multiparous women. The concentrations of albumin, prealbumin, and retinol-binding protein were correlated with birth weight, fetal growth retardation, and other measures of nutritional status. RESULTS: Serum albumin levels at 18 weeks correlated inversely with birth weight (P = .05). This negative correlation was explained by an inverse relationship between albumin concentration and maternal body mass index (BMI), and disappeared in a regression analysis adjusting for BMI. There was no significant correlation between albumin levels at 30 weeks and birth weight or between birth weight and the concentrations of the other two proteins at either gestational age. In individual subjects, the concentration of each protein correlated significantly with the concentration of the other proteins, and the levels at 18 weeks correlated with those at 30 weeks. CONCLUSION: Serum protein levels are not predictive of birth weight or growth retardation at birth, but do correlate significantly with a number of other measures of nutritional status. PMID- 8423942 TI - The effect of artificial caput on performance of vacuum extractors. AB - OBJECTIVE: To isolate and describe the effect of chignon formation (vacuum induced caput) on the performance of six vacuum extractors. METHODS: Six vacuum extractors were examined in the laboratory: the 6-cm Malmstrom, Mityvac, M-Type, Silc, Silastic, and Tender-Touch. The devices were tested in two model systems that duplicated the fetal scalp with and without chignon. A fetal cephalic model mimicked the head without chignon. The second model used the hind quarters of anesthetized dogs. Chignon was formed by slowly increasing vacuum until the target vacuum was reached. At increments of vacuum, traction was applied in the axis of the cup and maximal tractive force (pop-off) was measured. RESULTS: A statistically unique regression line described each device. There was no significant difference in the regression lines for the Malmstrom and M-Type cups with or without chignon. At equal vacuum, there was a significant decrease (P < .05) in maximal tractive force with chignon formation in the bell-shaped cups. CONCLUSIONS: There is a demonstrable deterioration in the performance of bell shaped cups with the formation of chignon. These data suggest that when using a bell-shaped extractor, efforts should be directed toward minimizing chignon formation by applying vacuum only with traction. PMID- 8423943 TI - Vibroacoustic-induced fetal movement: two stimuli and two methods of scoring. AB - OBJECTIVE: To determine whether the fetal movement response elicited by vibroacoustic stimulation depends upon the vibrator and the method used to judge movement. METHODS: Two methods of obtaining elicited fetal movement responses- maternal perceptions and ultrasound scan observations--were compared using two different vibroacoustic stimuli in 16 low-risk term pregnancies. RESULTS: Analyses of response over trials showed that the percentage of agreement between ultrasound scan observations and maternal perceptions varied (52-96%) across stimuli. The mothers perceived 64% less movement when vibrator 1 was used and 14% less movement response with vibrator 2. Analyses of average movement scores across subjects showed similar results. Vibrator 1 elicited significantly fewer maternal perceptions of fetal movement compared with ultrasound scan observation (an average of 0.8/3 compared with 2.3/3 movements per subject, respectively). For vibrator 2, there were no differences in average movement scores obtained by maternal perception and ultrasound scan observation (an average of 2.4/3 versus 2.8/3, respectively). CONCLUSION: The stimulus used in vibroacoustic stimulation testing influences the reliability of maternal movement perceptions as compared with ultrasound scan observations. PMID- 8423944 TI - Vibroacoustic stimulation evokes human fetal micturition. AB - OBJECTIVE: To examine the effect of vibroacoustic stimulation on fetal voiding as a measure of stress. METHODS: Fetal bladder volumes were examined serially by ultrasound 5 and 1 minutes before and 1 and 5 minutes after vibroacoustic stimulation (21 cases) or sham stimulation (20 cases). RESULTS: In the stimulated group, the mean (+/- SD) bladder volume decreased from 21.7 +/- 11.3 mL 1 minute before stimulation to 12.8 +/- 9.4 mL 1 minute after vibroacoustic stimulation, a statistically significant difference (P < .001). There was no significant decrease in bladder volume in the sham-stimulated fetuses. The correlations between bladder volume 1 minute before and 1 and 5 minutes after vibroacoustic stimulation were statistically significant (P < .01 and P < .001, respectively). CONCLUSION: Vibroacoustic stimulation induces fetal voiding, which may reflect a response to stress. PMID- 8423945 TI - Fetal vibroacoustic stimulation test: vibrator response characteristics in pregnant sheep postmortem. AB - OBJECTIVE: To determine the effect of small decreases in battery voltage on the sound pressure level and spectral features of sound within the uterus during vibroacoustic stimulation with a battery-powered electronic artificial larynx. METHODS: In eight pregnant, recently sacrificed sheep, the fluid-borne, intra amniotic sound pressures resulting from vibration of the abdominal wall with an electronic artificial larynx were compared with airborne sound pressures produced by an artificial larynx and measured at 50 cm. RESULTS: At all frequencies, the intrauterine sound pressure was higher than the sound pressure in air. Decreasing battery voltage from 9 to 7 V resulted in a 5-18-dB decrease in intrauterine sound pressure at the electronic artificial larynx fundamental frequency and most overtones, but only a 4-8-dB decrease in the airborne signal. The results indicate that for frequencies relevant to fetal hearing, the reduction of sound pressure level caused by inadequate battery voltage may result in a difference in fetal response. CONCLUSION: Routine verification of the function of a vibroacoustic device by means of a battery voltage check is recommended. PMID- 8423946 TI - Fetal serum and amniotic fluid magnesium concentrations with maternal treatment. AB - OBJECTIVE: To determine the effect of maternal intravenous (IV) magnesium sulfate administration on fetal serum and amniotic fluid (AF) concentrations of magnesium. METHODS: Thirty-six patients underwent fetal blood sampling for prenatal diagnosis of one of several abnormal conditions. Fifteen subjects had uterine contractions at the time of funipuncture and received IV magnesium sulfate therapy to quiet the uterus before the procedure. Three groups of subjects were defined: 21 untreated controls, ten women who received magnesium treatment for 1 hour, and five who received it for 3 hours before fetal blood sampling. Magnesium concentrations in maternal and fetal serum and AF were compared among the three groups. RESULTS: Patients who received magnesium sulfate for 1 and 3 hours had significantly higher concentrations of serum magnesium (4.11 +/- 0.6 and 5.54 +/- 0.2 mg/dL, respectively) than untreated subjects (1.77 +/- 0.2 mg/dL) (P < .0001). Fetal serum magnesium concentrations were significantly higher in the 1- and 3-hour groups (2.48 +/- 0.1 and 4.44 +/- 0.2 mg/dL, respectively) than in controls (1.67 +/- 0.2 mg/dL) (P < .0001). The AF magnesium concentration was significantly increased only after 3 hours of elevated maternal magnesium levels (1.45 +/- 0.2 and 2.84 +/- 0.2 mg/dL in controls and 3-hour group, respectively; P < .0001). The correlation between maternal and fetal blood magnesium concentrations was highly significant (r = 0.89; P < .0001), as was the correlation between fetal serum and AF concentrations (r = 0.84; P < .0001). CONCLUSION: Magnesium levels increase in fetal serum within 1 hour and AF within 3 hours after maternal IV administration. PMID- 8423947 TI - Osteocalcin levels in maternal and cord blood. AB - OBJECTIVE: To measure osteocalcin levels in fetal and maternal blood. METHODS: Osteocalcin was measured by radioimmunoassay in paired maternal and umbilical venous and arterial blood obtained at term deliveries. RESULTS: Umbilical venous osteocalcin levels (mean +/- standard deviation) were significantly higher than arterial levels (10.28 +/- 4.99 versus 3.85 +/- 2.27 micrograms/L). Umbilical venous but not arterial levels of osteocalcin were significantly higher than maternal values (2.54 +/- 1.19 micrograms/L). CONCLUSION: Higher osteocalcin levels in umbilical venous blood than in umbilical arterial blood suggest that the placenta may be the main source of osteocalcin in late fetal life. PMID- 8423948 TI - Alterations in maternal-fetal Doppler flow velocity waveforms in preterm labor patients undergoing magnesium sulfate tocolysis. AB - OBJECTIVE: To evaluate the effects of magnesium on fetal hemodynamics in mothers treated with magnesium sulfate for preterm labor. METHODS: Twenty women who presented to the Carolinas Medical Center in preterm labor between 24-35 weeks' gestation were enrolled in the study. Pulsed-wave Doppler measurements were made from the fetal middle cerebral artery, the fetal umbilical artery, and the maternal uterine arteries before the initiation of magnesium sulfate tocolysis and during its intravenous administration. Baseline measurements were compared with intra-therapy measurements using paired t tests. RESULTS: During the administration of magnesium sulfate, the fetal middle cerebral artery diastolic blood flow velocity decreased significantly (P = .05) and the maternal uterine artery diastolic blood flow velocity increased significantly (P = .01). There were no significant changes in the umbilical artery diastolic blood flow measurements. These findings remained the same after adjusting for gestational age, estimated fetal weight, frequency of contractions, serum magnesium level at the time of the intra-therapy Doppler study, and maternal blood pressure. CONCLUSION: The alterations in fetal hemodynamics during magnesium sulfate administration suggest a physiologic normalization process related to the stressed preterm infant and the preterm labor process itself. PMID- 8423949 TI - An obstetric scoring system: its development and application in obstetric management. AB - OBJECTIVE: To develop a statistically derived but clinically usable antenatal risk scoring system. METHODS: Data from 20,985 pregnancies were statistically analyzed to identify significant risk factors. Logistic regression analysis was then used to produce a final scoring system, which was subsequently tested for validity on a separate population of 3120 pregnancies. RESULTS: Twenty-seven significant antenatal variables were included in the final scoring system. Application of the system in early pregnancy resulted in a predictive accuracy of 0.73; at the onset of labor, predictive accuracy was 0.91. At the time of labor, 87% of poor outcomes were accurately identified by allocation of only 16% of the women to the high-risk group. CONCLUSIONS: It was possible to develop a risk scoring system with a predictive accuracy higher than any previously reported statistically derived score. Summation of the logistic coefficients provides a score that by comparison with a chosen threshold identifies a high-risk pregnancy. In this way, despite the complexity of statistical analysis, all clinicians can quickly apply this scoring system. PMID- 8423950 TI - Maternal age: an independent risk factor for cesarean delivery. AB - OBJECTIVE: To evaluate whether there is an increased risk of cesarean delivery in women who delay childbearing into their later reproductive years, using a well defined cohort and controlling for difference in baseline susceptibility such as complications of pregnancy and labor. METHODS: The current study included all women at least 20 years of age delivering their first child and evaluated between 1988-1991 as part of the prospective Yale Health in Pregnancy Study (N = 735). All women planned to deliver at Yale-New Haven Hospital. The study group was divided into three maternal age groups: 20-29 (N = 422), 30-34 (N = 239), and 35 and over (N = 74). Information collected included demographic characteristics, medical history, pregnancy complications, labor complications, mode of delivery, and pregnancy outcome. RESULTS: The cesarean delivery rates for the three age groups were 16.8% (71 of 422), 26.8% (64 of 239), and 32.4% (24 of 74), respectively. This trend was highly significant when evaluated with chi 2 for linear trend (P = .0002). Within groups of women with and without complications of pregnancy or labor, cesarean rates increased with maternal age. Using stepwise logistic regression to adjust simultaneously for confounding variables, adjusted odds ratios were 1.6 (95% confidence interval [CI] 0.9-2.7) for the women aged 30 34 and 2.3 (95% CI 1.1-4.8) for the 35 and older age group. CONCLUSIONS: Maternal age appears to be an independent risk factor for cesarean delivery. The reasons for this clinically important and statistically significant increased risk are unclear, but may be due to physician and patient concern over pregnancy outcome in older women. PMID- 8423951 TI - The perioperative morbidity of scheduled cesarean hysterectomy. AB - OBJECTIVE: To compare the outcomes of patients undergoing scheduled cesarean hysterectomy with those of women treated with cesarean delivery and subsequent hysterectomy. METHODS: Through a retrospective review of 43 patients, we investigated the morbidity associated with scheduled cesarean hysterectomy and compared these findings with the combined morbidity of scheduled cesarean delivery and subsequent abdominal hysterectomy in a control population. Controls were included only if the subsequent hysterectomy was performed within 3 years of the index cesarean delivery. Each study subject was assigned two controls matched for age, parity, number of previous cesarean deliveries, and indications for procedures. The incidence of the following major morbidity events was compared between the groups: transfusion, urinary tract injury, fistula formation, cellulitis or endometritis, postoperative abscess or hematoma formation, ileus, pneumonia, and wound complications requiring prolonged therapy (seroma, hematoma, infection). RESULTS: The number of women receiving transfusions after scheduled cesarean hysterectomy was greater than among controls (39.5 versus 15.1%; P < .05). The proportion of patients with major morbidity, exclusive of transfusion, was significantly greater in the control population (44%) than in women with scheduled cesarean hysterectomy (16%) (P < .05). The cumulative number of women with a major complication, such as transfusion or a morbid event, was 22 of 43 in the study group versus 44 of 86 in the control population, a nonsignificant difference. CONCLUSION: We found no significant difference in the cumulative perioperative complication rates in women undergoing scheduled cesarean hysterectomy compared with a population of similar patients treated with cesarean delivery and subsequent abdominal hysterectomy. PMID- 8423952 TI - Fundal height as a predictor of preterm twin delivery. AB - OBJECTIVE: To determine whether fundal height might predict early delivery in twin pregnancies; that is, whether larger fundal heights predispose to earlier delivery. METHODS: From the charts of 336 well-dated twin pregnancies, we generated a series of fundal height curves. RESULTS: Within our population of twin pregnancies, the mean fundal height at any given gestational age did not differ between pregnancies delivered before 34 weeks and those delivered at or after 34 weeks. A single fundal height measurement above the 90th percentile before 34 weeks yielded a sensitivity of 23% and a specificity of 79% for delivery before 34 weeks, with a positive predictive value of 38% and a negative predictive value of 64%. CONCLUSION: Factors other than uterine overdistention (as measured by fundal height) must be implicated in preterm twin delivery. PMID- 8423953 TI - Cervical dilation: accuracy of visual and digital examinations. AB - OBJECTIVE: To determine the accuracy of visually versus digitally determined cervical dilation. METHODS: We studied 64 pregnant women with rupture of the amniotic membranes. One of two residents performed a speculum examination, estimated the dilation and effacement visually, and noted whether fetal parts or amniotic membranes were present. The same examiner then determined digitally the cervical dilation and effacement and station of the presenting part. RESULTS: There was a statistically significant correlation between visually and digitally determined cervical dilation (r = 0.78, P < .001). CONCLUSION: Visual estimation of cervical dilation correlates well with digital examination. PMID- 8423954 TI - Changed proteoglycan metabolism in human cervix immediately after spontaneous vaginal delivery. AB - OBJECTIVE: To investigate whether the final cervical ripening just before or during labor corresponds to changes in cervical connective tissue metabolism and composition. METHODS: Cervical biopsies taken transvaginally immediately after normal vaginal delivery and at hysterectomy were incubated with [35S] sulfate. In other biopsy specimens, the nonlabeled proteoglycan pattern was studied. Collagen concentration and organization were monitored to estimate the degree of cervical ripeness. Labeled and nonlabeled proteoglycans were isolated, characterized, and quantitated. Collagen concentration and collagen, extractable and not extractable by pepsin digestion, were quantitated as hydroxyproline. RESULTS: A marked increase in the accumulation rate of labeled proteoglycans was observed in cervical biopsies obtained immediately after vaginal delivery. This increase was markedly larger than that previously noted in term pregnant women with unfavorable cervices. In particular, the accumulation of one labeled large chondroitin sulfate proteoglycan and one small dermatan sulfate proteoglycan, biglycan, increased significantly compared with the unfavorable cervices. In contrast, the small dermatan sulfate proteoglycan, decorin, showed a relative decrease. The changed synthesis of proteoglycans resulted in a two- to threefold increase in chemical amounts of the large proteoglycan, while the small dermatan sulfate proteoglycans decreased to approximately half of that in the nonpregnant cervix. CONCLUSION: The changed cervical proteoglycan metabolism results in a changed proteoglycan composition of the tissue. This alteration probably influences collagen organization, contributing to the cervical softening and dilatation essential for an uncomplicated vaginal delivery. PMID- 8423955 TI - Glove perforations and blood contact associated with manipulation of the fetal scalp electrode. AB - OBJECTIVE: To assess prospectively the frequency of glove injury associated with insertion of the fetal scalp electrode and subsequent examination of the cervix with the electrode in place. METHODS: Over a 7-month period, sterile gloves were collected after use for insertion of the fetal scalp electrode or cervical examination with the electrode in place. Attendants indicated their level of training, time, date and purpose of glove use, and cervical examination. They also noted whether they were aware of a glove perforation or observed blood, amniotic fluid, or genital tract secretions on their hand. Glove patency was assessed by filling the glove with water to 1.5-2.0 times its normal volume and observing for leaks. One hundred unused gloves were tested for patency and served as controls. RESULTS: Five hundred one gloves were evaluated, of which 13 (2.6%, 95% confidence interval [Cl] 1-4%) had perforations. Seven of 277 gloves (2.5%) used only for examinations had perforations, compared with six of 244 (2.5%) used only for insertion of the electrode. Two percent (95% CI 0-5%) of the unused control gloves had perforations. These observed differences were not statistically significant. Nineteen attendants (3.8%, 95% CI 2.1-5.5%) noted blood or genital tract secretions on their hand after insertion of the electrode (N = 4) or subsequent cervical examination (N = 15). Only one point of contract resulted from a glove perforation; the other 18 were on the wrist and apparently resulted from leakage of fluid around the open cuff of the glove. CONCLUSIONS: The risk of glove perforation during insertion of the fetal scalp electrode or subsequent cervical examination is low if proper technique is observed. Blood or fluid contact is more likely to result from leakage of fluid around the open cuff of the glove during a vaginal examination. PMID- 8423957 TI - Maternal plasma D-dimer levels in normal and complicated pregnancies. AB - OBJECTIVE: To evaluate D-dimer as a marker for fibrinolysis in normal and complicated pregnancies using an enzyme-linked immunosorbent assay (ELISA) technique. METHODS: Four groups of pregnant women were enrolled: 17 normal women followed longitudinally from 28-40 weeks' gestation, 14 patients with preterm labor at 28-34 weeks, 17 patients with preeclampsia at term (37-40 weeks), and 14 patients with abruptio placentae (32-40 weeks). We assayed peripheral venous blood samples from each patient for D-dimer levels using a commercial ELISA kit. D-dimer values were calculated by regression analysis using internal standards and controls for each assay. Data were compared using Student t test or analysis of variance with repeated measures. RESULTS: D-dimer values increased slightly with increasing gestational age. Patients with preterm labor, preeclampsia, and abruptio placentae had mean D-dimer values significantly greater than those of controls (P < .003). D-dimer values of the abruption group were approximately twice those of the control group (3393 +/- 2086 versus 1750 +/- 839 ng/dL). CONCLUSION: An increase in fibrinolysis may be associated with the pregnancy complications studied, as reflected by alterations in maternal plasma D-dimer levels. PMID- 8423956 TI - Pulmonary edema associated with pregnancy: echocardiographic insights and implications for treatment. AB - OBJECTIVE: To evaluate the role of echocardiography in determining the cause of pulmonary edema in pregnancy and the impact this information has on management. METHODS: We studied prospectively 45 pregnant or recently postpartum women admitted to an obstetric intensive care unit with pulmonary edema during a 6-year period. Between 1 and 4 days after the onset of pulmonary edema, two-dimensional and M-mode echocardiography was performed, as was continuous, pulsed, and color Doppler echocardiography. The clinical diagnosis obtained from history, physical examination, chest radiograph, and laboratory data was compared with the echocardiographic diagnosis. RESULTS: Three therapeutically and prognostically distinct groups were identified by echocardiography: 1) those with decreased systolic function (N = 19), 2) those with normal systolic function but increased left ventricular mass and presumed diastolic dysfunction (N = 17), and 3) those with normal hearts (N = 9). During the study period, two patients with systolic dysfunction died and one underwent cardiac transplantation. Patients with systolic dysfunction required short- and long-term treatment with digoxin, diuretics, and angiotensin-converting enzyme inhibitors. Those with diastolic dysfunction received diuretics and long-term antihypertensive therapy. Women with normal hearts required acute therapy only. In 21 patients (47%), echocardiography demonstrated clinically unsuspected findings, which altered the long-term management in 16. CONCLUSION: Because clinical and roentgenographic findings do not accurately differentiate patients with respect to the presence and type of cardiac dysfunction, and because these subgroups differ with respect to treatment and probably prognosis, we recommend echocardiography to evaluate all pregnant women with pulmonary edema. PMID- 8423958 TI - The prevalence of substance abuse among pregnant women in Utah. AB - OBJECTIVE: To determine the rate of substance use among Utah's pregnant women, who are primarily white and middle class, for comparison to rates reported in studies of inner-city populations. METHODS: Urine specimens and demographic data were obtained anonymously from women who delivered infants at ten hospitals in urban and suburban Utah. Urine samples were screened by enzyme immunoassay for amphetamines, marijuana, cocaine, methadone, opiates, benzodiazepines, and ethanol. RESULTS: Among 792 women screened, the mean age was 26.2 years, 86.1% were white, 62.9% were multigravidas, and 66.3% had private insurance. Cocaine was detected in nine samples (1.1%), illicit amphetamines in five (0.6%), marijuana in 23 (2.9%), ethanol in 32 (4.0%), over-the-counter amphetamines in 51 (6.4%), and benzodiazepines in seven (0.9%). The prevalence rate for women positive for illicit drugs and alcohol combined was 7.8%. Cocaine-positive and marijuana-positive women were more likely to be non-white or Hispanic and to have Medicaid or no insurance than were women negative for either substance. Women with Medicaid or no insurance were four times more likely to be positive for illicit substances (10.7%) than were those with private insurance (2.3%) (P = .0001). CONCLUSION: The rates and patterns of substance use differ between Utah's pregnant women and inner-city populations. The patterns in Utah may be more representative of many communities in the United States that have a predominantly middle-class, white population. PMID- 8423959 TI - Systematic serologic screening for toxoplasmosis in pregnancy. AB - OBJECTIVE: To determine which serologic method or combination of methods could most effectively identify acute toxoplasmosis during pregnancy and to assess whether systematic screening is practical and cost-effective. METHODS: Using basic serologic tests (direct agglutination test and immunosorbent agglutination assay), we screened 2104 women for toxoplasmosis. Immunoglobulin M (IgM)-reactive patients (IgM immunosorbent agglutination assay score of at least 3) were studied by differentiating serologic tests performed sequentially. RESULTS: Specific immunity was found in 874 pregnant women (41.6%); 155 (7.4%) were IgM-reactive and 12 (0.6%) had acute toxoplasmosis. Using a reduced immunosorbent agglutination assay score of at least 3 (normally a score of at least 6 is used), acute toxoplasmosis was identified in 11 women at their first prenatal visit, including two in whom acute infection would not have been detected by a score of 6 or more until 9 weeks later. One additional nonimmune patient with acute infection was identified only by follow-up serologic testing. CONCLUSION: Systematic screening followed by sequential differentiating serologic tests is practical and cost-effective for the diagnosis of acute toxoplasmosis during pregnancy. PMID- 8423960 TI - Can prenatal vitamin K1 (phylloquinone) supplementation replace prophylaxis at birth? AB - OBJECTIVE: To assess the effect of prenatal vitamin K1 on the coagulation status of newborns. METHODS: We measured noncarboxylated prothrombin and performed the Normotest in two groups of 5-day-old infants whose mothers were given oral vitamin K1, 10 mg/day for 2 weeks at least 10 days before delivery, or were untreated. RESULTS: Noncarboxylated prothrombin was found in one of 74 treated women and 13 of 186 controls, a nonsignificant difference. The mean (+/- standard deviation) Normotest value was 59.6 +/- 10.1% (range 38.9-84.4) for the treated group and 53.4 +/- 9.9% (range 16.3-89.9) for the controls, a statistically significant difference (P < .001). CONCLUSION: Based on the Normotest results, we suggest that vitamin K crosses the placenta and persists to activate the vitamin K-dependent coagulant factors until at least the fifth day of life. Thus, prenatal vitamin K1 administration may replace prophylaxis at birth. PMID- 8423961 TI - Achievement of therapeutic concentrations of cefuroxime in early preterm gestations with premature rupture of the membranes. AB - OBJECTIVE: To determine whether therapeutic cefuroxime concentrations can be achieved in maternal plasma, amniotic fluid (AF), neonatal plasma, placenta, and membranes in women with premature rupture of the membranes (PROM) at 27-33 weeks' gestation. METHODS: In an open nonrandomized, dose-response study, nine patients with PROM at 27-33 weeks' gestation received 1.5 g of cefuroxime intravenously three times daily. Maternal plasma and AF specimens were collected during pregnancy, and umbilical cord plasma, placenta, and membrane specimens were collected after delivery to assay cefuroxime concentrations using high performance liquid chromatography. RESULTS: A high rate of placental transfer of cefuroxime was found. Bactericidal concentrations could be demonstrated in maternal plasma and in AF leaking from the vagina. A concentration-time curve in AF could be detected, with a peak concentration 3-4 hours after infusion. Therapeutically active levels were present in the newborns. The resorption of cefuroxime by the fetal membranes was high. CONCLUSIONS: Therapeutic concentrations of cefuroxime were found in all body fluids and tissues studied. Further study to establish the clinical utility of cefuroxime prophylaxis in PROM seems appropriate. PMID- 8423962 TI - Total and regional bone mass values and biochemical markers of bone remodeling in endometriosis. AB - OBJECTIVE: TO measure peripheral, axial, and total bone mass and to assess markers of bone remodeling in women with endometriosis, with the aim of addressing previous reports of diminished peripheral bone mass in these patients. METHODS: Whole body bone densitometry, estradiol (E2) levels, and biochemical bone markers (calcium, phosphorus, total alkaline phosphatase, tartrate-resistant acid phosphatase, and total proteins) were determined in 28 patients with endometriosis and compared with findings in 33 controls. RESULTS: There were no significant differences between the groups in bone mass at different sites or in biochemical bone markers. We observed a significant negative correlation between tartrate-resistant acid phosphatase and E2 levels (P < .001) and with total (P < .001), head (P < .01), and axial (trunk) (P < .001) bone mass. Total alkaline phosphatase did not correlate with any of the indices studied. CONCLUSIONS: Bone mass was not lower in any of the areas studied in women with endometriosis. There was a significant negative correlation of tartrate-resistant acid phosphatase with E2 and with total, head, and trunk bone mass. PMID- 8423963 TI - Platelet-activating factor acetylhydrolase activity in peritoneal fluids of women with endometriosis. AB - OBJECTIVE: To investigate whether platelet-activating factor acetylhydrolase may play a role in the pathophysiology of endometriosis. METHODS: Peritoneal fluids were obtained at laparoscopy from eight fertile women without endometriosis and 12 women with mild endometriosis. Platelet-activating factor acetylhydrolase activity was assayed in serum and peritoneal fluid by the release of 3H-acetate from [3H]-platelet-activating factor during a 15-minute incubation. RESULTS: Platelet-activating factor acetylhydrolase activity (mean +/- standard error of the mean) was significantly lower (P < .01) in peritoneal fluids from women with mild endometriosis (29.5 +/- 2.7%) than in controls (39.9 +/- 3.4%). The platelet activating factor acetylhydrolase-cholesterol and -albumin ratios were also significantly lower in peritoneal fluids of women with endometriosis compared with controls (P < .006 and P < .012, respectively). CONCLUSION: These data suggest that mild endometriosis is associated with decreased platelet-activating factor acetylhydrolase activity in peritoneal fluid. PMID- 8423964 TI - The effect of educational brochures on knowledge and emotional distress in women with abnormal Papanicolaou smears. AB - OBJECTIVE: To determine whether an educational brochure about abnormal Papanicolaou smears could reduce psychological distress and concerns about cancer and future health, and increase knowledge about dysplasia and the investigation, treatment, and recommended follow-up. METHODS: One hundred twenty-five consecutive women with dysplasia on a Papanicolaou smear referred to a colposcopy clinic for their first examination were alternately assigned to receive a mailed educational brochure the same day or to receive no brochure. At the colposcopy clinic, consenting women were given the Brief Symptom Inventory, specific questions about feelings regarding an abnormal smear, and a knowledge quiz about dysplasia, colposcopy, and the recommended follow-up. RESULTS: Women who received the brochure were significantly less distressed on the Brief Symptom Inventory and were less anxious about the abnormal Papanicolaou smear, the fear of cancer, and their future health than were women not receiving the brochure. Study women also performed significantly better on knowledge questions about dysplasia, colposcopy, and the recommended follow-up. CONCLUSION: Given the low cost, ease of delivery, and usefulness of educational brochures in reducing psychological distress and increasing knowledge specific to the condition, an educational brochure should routinely accompany or immediately follow notification about an abnormal Papanicolaou smear. PMID- 8423965 TI - Pelvic muscle exercises: when do they work? AB - OBJECTIVE: To identify urodynamic indices that can predict the outcome of pelvic muscle exercises in women with genuine stress urinary incontinence. METHODS: Thirty-six women with genuine stress urinary incontinence were evaluated during a 6-month period. Urodynamic studies were performed before and 3 months after completion of a program of Kegel pelvic muscle exercises. Subjective symptoms and objective loss of urine as well as pad count were used to evaluate the outcome of the exercise program. A receiver-operating characteristic curve was used to assess the prognostic value of the pressure transmission ratio between the abdomen and urethra. RESULTS: Twenty patients (56%) were cured or substantially improved 3 months after completing the pelvic muscle training, whereas 16 were unchanged. The urethral closure pressure significantly increased in the subjects cured or improved. After successful training, significant changes were noted in the pressure transmission ratio between the abdomen and urethra on cough. Six of seven subjects with mild incontinence responded favorably, whereas 13 of 15 with severe incontinence did not improve after the training. The pressure transmission ratio plotted on a receiver-operating characteristic curve was found to have optimal predictive value at the 80% level. CONCLUSION: Kegel pelvic muscle exercises give a better outcome in women with mild stress urinary incontinence and/or with a pressure transmission ratio between the abdomen and urethra of 80% or more. PMID- 8423966 TI - Ectopic pregnancy in the United States: economic consequences and payment source trends. AB - OBJECTIVE: To estimate the annual direct and indirect costs of ectopic pregnancy in the United States and to examine trends in payment source. METHODS: We analyzed hospital discharge data for 1982-1989 from a San Francisco hospital and California statewide data for 1983-1987 to estimate direct medical care costs and to determine payment sources. Examination of national labor data was used to compute indirect cost. RESULTS: The total cost of ectopic pregnancy in 1990 was estimated to be nearly $1.1 billion. Direct costs of hospitalization and other medical treatment contributed 77% of the total costs, with average hospital costs estimated to be $6079, hospitalization-related physician fees $3254, and average outpatient costs $149, for a total direct cost-per-case of $9482. Of the total indirect costs ($250.5 million), 67% was attributed to the value of lost wages and the remainder to the lost value of household management. Public payment sources covered the largest portion of ectopic pregnancy-related direct costs among women aged 19 and younger (35%), but private insurance covered the largest portion among women aged 20-29 (32%) and women aged 30 and older (43%). In general, the proportion of payments made by private insurance has decreased, while the proportion of payments made by public pay sources and health maintenance organizations and preferred provider organizations has increased. CONCLUSIONS: Ectopic pregnancy results in a substantial economic burden, with an increasing share of direct costs being borne by public pay sources. Appropriate use of cost-effective management approaches can reduce costs, while preventive measures that decrease the risk of ectopic pregnancy can both save resources and, more important, spare human suffering. PMID- 8423967 TI - Acetic acid visualization of the cervix to detect cervical dysplasia. AB - OBJECTIVE: To determine whether acetic acid visualization of the cervix can identify cervical dysplasia and improve detection of lesions missed by Papanicolaou test screening. METHODS: During a 2-year period, patients attending family planning clinics for regular gynecologic examinations had acetic acid applied to the cervix, followed by gross visualization without magnification. Patients with suspicious acetowhite lesions and normal Papanicolaou tests were referred for colposcopic evaluation. Findings from these examinations and corresponding biopsy results were analyzed retrospectively. RESULTS: Eighty-five women were referred for colposcopy because of abnormal acetowhite areas on the cervix. Thirteen patients (15%) had cervical intraepithelial neoplasia (CIN) (nine CIN I, four CIN II), 22 (26%) had koilocytosis, and 16 (19%) had benign histologic findings. In total, 51 patients had suspicious lesions at colposcopy for which biopsies were performed, and 34 (40%) had normal colposcopic examinations. CONCLUSIONS: Acetic acid visualization of the cervix can detect dysplasia otherwise missed by Papanicolaou test screening. However, further refinements in technique are required to decrease false-positive findings and unnecessary referrals for colposcopy. PMID- 8423968 TI - Clinical implications of tumor volume measurement in stage I adenocarcinoma of the cervix. AB - OBJECTIVE: To evaluate the prognostic significance of three-dimensional determination of tumor size in stage I cervical adenocarcinoma. METHODS: Tumor volume was measured using hematoxylin and eosin-stained sections of cone biopsy and hysterectomy specimens from 36 patients with stage I adenocarcinoma of the cervix. The volume was then correlated with pelvic lymphatic spread and clinical outcome. RESULTS: The subjects were followed for a mean (+/- SEM) of 63 +/- 8 months. No recurrence or lymphatic seeding was encountered in the 22 tumors measuring no more than 500 mm3. Two of 25 tumors (8%) having up to 5 mm depth of stromal invasion had lymph node metastasis, one of which was 1.5 mm, compared with four of 11 (36%) in the group with deeper than 5 mm invasion (P < .02). The depth of stromal invasion predicted recurrence less significantly. Among the 25 tumors with up to 5 mm stromal invasion, two recurred, compared with three of 11 with more than 5 mm invasion (P < .1). Two women who had tumor volumes below 500 mm3 and depths of stromal invasion up to 8.5 mm were disease-free at 52 and 96 months of follow-up. On the other hand, tumors with 2.6 and 3.8 mm stromal invasion, but with volumes exceeding 500 mm3, recurred. CONCLUSION: Tumor volume is a better predictor of pelvic lymph node metastasis and recurrence than is the depth of stromal invasion in stage I cervical adenocarcinoma. PMID- 8423970 TI - Survey of forceps delivery in North America in 1990. AB - OBJECTIVE: To determine whether opinions and practice patterns have markedly changed over the past decade and whether clinicians are cognizant of the new ACOG definitions for forceps deliveries. METHODS: Two hundred ninety-five United States and Canadian residency programs were surveyed via a questionnaire; 203 (69%) responded. Statistical analysis was performed using the chi 2 test, Yates correction factor, and the extended Mantel-Haenszel test. RESULTS: All but two program directors (99%) were familiar with the new definitions, which were being used by 162 (80%) of the programs. All institutions used outlet and low forceps, although 14% no longer performed midforceps deliveries. Attending faculty were the primary instructors in 67% of United States and 100% of Canadian programs. Simpson forceps were the most commonly used for outlet (46%) and low (43%) forceps deliveries. Kielland (27%) and Simpson (24%) instruments were most commonly used for midforceps deliveries. CONCLUSION: Although the rate of midforceps use decreased, operative vaginal delivery was still commonly taught in residency programs in North America in 1990. PMID- 8423969 TI - Outpatient abortion for fetal anomaly and fetal death from 15-34 menstrual weeks' gestation: techniques and clinical management. AB - OBJECTIVE: To determine the safety of providing outpatient abortion services for women with complicated advanced pregnancies. METHODS: During a 10-year period, 124 abortions were performed after 14 menstrual weeks' gestation at an outpatient abortion facility for indications of fetal anomaly, diagnosed genetic disorder, or fetal death. Gestational lengths ranged from 15-34 menstrual weeks. Fetal diagnoses included a variety of chromosomal abnormalities, malformations, and death. Techniques for performing the late abortions included a serial multiple laminaria method of cervical dilation. Abortions performed after 20 menstrual weeks were effected by instillation of intra-amniotic hyperosmolar urea or induction of fetal death by injection of digoxin and/or hyperosmolar urea into the fetus, followed by artificial rupture of membranes, induction of labor, and assisted expulsion or instrumental extraction of the fetus. At less than 20 weeks, dilation and evacuation following serial multiple laminaria treatment of the cervix was the method of choice. RESULTS: The median gestational age was 23 menstrual weeks. The median procedure time for all cases was 12 minutes and median blood loss was 125 mL. Procedure time increased with length of gestation (P = .00). Blood loss was only slightly increased by gestation length (P = .154) and not by procedure time (P = .299). Complication rates were not significantly related to gestation length (P = .895). There was one major complication in this series. There were no uterine perforations and one cervical laceration. CONCLUSION: Outpatient abortion may be performed safely in most cases of fetal disorder, including death, through 34 menstrual weeks under proper conditions. PMID- 8423971 TI - Residency training in contraception, sterilization, and abortion. AB - Obstetrics and gynecology residency program directors and chief residents were surveyed, and 80% responded to a questionnaire regarding training in contraception, sterilization, and abortion. Most program directors reported that teaching and clinical experience is provided in these areas. The chief residents, however, reported having less clinical experience than was estimated by their program directors. Most residents reported experience prescribing oral contraceptives and performing tubal ligations, but had little experience with other methods. Thirty-eight percent of graduating chief residents reported never having inserted an intrauterine device, 47% had never performed a first-trimester abortion, and 43% had never performed a dilation and evacuation. Residency programs need to set competency goals in these areas so that new practitioners will be well prepared to care for American women who wish to control their fertility. PMID- 8423972 TI - Should women with familial ovarian cancer undergo prophylactic oophorectomy? PMID- 8423973 TI - Epidural analgesia in labor and fetal hyperthermia. PMID- 8423974 TI - Clinicians who provide abortions: the thinning ranks. PMID- 8423975 TI - Clinicians who provide abortions: the thinning ranks. PMID- 8423976 TI - Quantitative and qualitative effects of douche preparations on vaginal microflora. PMID- 8423977 TI - Quantitative and qualitative effects of douche preparations on vaginal microflora. PMID- 8423978 TI - [Complex evaluation of risk factors in patients after myocardial infarction]]. AB - In this work the authors have constructed a clinical epidemiological score system for characterizing the severity of the diseased state of our 85 patients got over myocardial infarction. The important role is commonly known which is played by the free radical processes in the pathogenesis of the heart infarct. From these free radical reactions the authors have focused their attention to studying the lipid peroxides. Beside the determinations of the malondialdehyde content of native serums they have measured the intensity of the malondialdehyde development in the free radical system generated by hydrogen-peroxide in vitro treatment of serums. With the help of the newly elaborated and applied method they have obtained a 67.11% average increase of the amount of malondialdehyde developed in vitro in comparison with the mean malondialdehyde value measured in the same analytical conditions with 154 healthy controls. They have made a statistical calculation for studying the question if there is any relationship between the scores of the age of patients and the malondialdehyde values of hydrogen-peroxide treated serums. On the basis of results of biochemical measurements and statistical analyses it seems very important to pay more attention both to the intensity of free radical reactions and the patients age in the severity's relations of the diseased state at heart infarct patients. PMID- 8423979 TI - [Occult breast cancer causing axillary metastasis]. AB - Authors review 43 cases of occult breast cancer selected from 6396 patients operated on for breast cancer in the National Institute of Oncology, Budapest over a 20-year period (1972-1991). In 20 patients mastectomy and axillary block dissection, in 9 upper-outer quadrant resection of the breast with axillary block dissection, in 12 only axillary block dissection, and in 2 axillary biopsy were carried out. The primary tumor was localized only in 11 patients during the first operation either by macroscopic or by microscopic examination. In 12 patients' breast the primary tumor became detectable only later. Out of the 20 mastectomy specimens, 2 primary tumors could not be identified even with the most thoroughful examination. Out of the breast conservation group in 4 patients, the primary tumor remained occult until recently, and in the axillary block dissection group (12 patients) the site of the primary cancer could be identified only in 2 patients. The authors deal with diagnostic difficulties and the dilemmas of the surgical indications and treatment. PMID- 8423980 TI - [Laparoscopic surgery in tubal pregnancy]. AB - The author accounts about twenty patients operated on tubal pregnancy established on ultrasonic and laparoscopy. He refers the history of the methods of surgical interventions and the difficulties of the diagnosis nowadays. He makes known the technics of the laparoscopical operation and the operated patients' postoperative status. PMID- 8423981 TI - [The role of CT scan in cases of injury to the inferior orbital muscle]. AB - Blowout fracture, involving the inferior wall of the orbit, causing the entrapment of the inferior rectus muscle is relatively rare, and is usually seen after traffic or sport accidents. By presenting two cases, the authors draw the attention to the necessity of careful CT scanning of the orbit, when after relatively slight trauma the patients presents with diplopia and inhibited movement of the eye. A correct investigation can establish the diagnosis and early surgery gives a full recovery of the symptoms. PMID- 8423982 TI - [Reestablishment of the Finnish-Hungarian-Estonian Medical Society]. PMID- 8423983 TI - [Beginning of a cooperation between physicians of the Finno-Ugric peoples]. PMID- 8423984 TI - [A Hungarian princess as "founder" of a modern European hospital?]. PMID- 8423985 TI - [Current questions concerning TV and medicine]. PMID- 8423986 TI - E mu-bcl-2 transgene facilitates spontaneous transformation of early pre-B and immunoglobulin-secreting cells but not T cells. AB - To assess the lymphoid tumorigenic potential of bcl-2, mice of five independent strains expressing a bcl-2 transgene in B and/or T cells were monitored for disease up to 12 months of age. Lymphoma prevalence was minimal in the T lineage but significant, although low (3-15%), in the B lineage. The principal types of tumors were plasmacytomas secreting immunoglobulin and novel lymphomas that expressed markers such as Sca-1, CD4, Thy-1, CD34 and CD45(B220), consistent with an origin very early in B-lymphoid development. Rearrangement of the c-myc gene was common in the plasmacytomas, implying a synergistic role for myc and bcl-2 in their etiology, but was not detected in the lymphomas. PMID- 8423987 TI - BCR-ABL tyrosine kinase is autophosphorylated or transphosphorylates P160 BCR on tyrosine predominantly within the first BCR exon. AB - The role of BCR gene sequences in Philadelphia (Ph) chromosome-positive leukemia is not well understood. Our previous studies demonstrated that P210 BCR-ABL co precipitates with P160 BCR following immunoprecipitation with antibodies to the C terminal domain of P160 BCR, sequences lacking in P210 BCR-ABL. We now report that tryptic peptides shared by both P160 BCR and P210 BCR-ABL are phosphorylated on tyrosine in vitro either when using immune complexes containing P160 BCR complexed to BCR-ABL or when P160 BCR is phosphorylated in trans by P210 BCR-ABL immune complexes from cells lacking functional P160 BCR. P185 BCR-ABL produced in a cell line derived from a Ph chromosome-positive acute lymphocytic leukemia patient also co-immunoprecipitated with P160 BCR. As with P210 BCR-ABL, P160 BCR tyrosine phosphopeptides were shared with P185 BCR-ABL, indicating that the major sites of tyrosine phosphorylation in vitro are contained within the first exon of P160 BCR. Similarly, BCR-ABL autophosphorylation was found to occur predominantly at tyrosines within BCR exon 1 sequences. These results raise the possibility that the activated ABL protein kinase of BCR-ABL proteins modulates the putative signal transduction activities of P160 BCR by tyrosine phosphorylation of exon 1 sequences. PMID- 8423988 TI - NYK/FLK-1: a putative receptor protein tyrosine kinase isolated from E10 embryonic neuroepithelium is expressed in endothelial cells of the developing embryo. AB - A protein tyrosine kinase (NYK/FLK-1), bearing all the hallmarks of a growth factor receptor, has been isolated from a cDNA library generated from enriched populations of mouse day 10 embryonic neuroepithelium and from day 18 embryonic colon. Sequence analysis of cDNAs covering the entire coding region of this 5.7 kb mRNA predicted the presence of seven immunoglobulin-like domains in the extracellular region of this molecule. This feature, coupled with the detection of an insert domain bisecting the kinase domain of the predicted protein sequence, places NYK/FLK-1 firmly in the Platelet-derived Growth Factor Receptor related class of molecules. NYK/FLK-1 is expressed at high levels in adult heart, lung, kidney, brain and skeletal muscle, but is also expressed at lower levels in most other adult tissues. In situ hybridization of day 12.5 embryo sections demonstrated NYK/FLK-1 mRNA expression in endothelial cells lining the dorsal aorta and intervertebral veins. In addition, expression was found in cells lining the capillary plexus which surrounds the developing neuroepithelium, and in the endothelial cells which are found within the embryonic lung, spleen, liver and metanephros. PMID- 8423989 TI - Methylation-sensitive DNA binding by v-myb and c-myb proteins. AB - The retroviral oncogene v-myb and its cellular homolog c-myb encode nuclear DNA binding phosphoproteins (v-MYB and c-MYB) that function as transcriptional regulators. v-MYB and c-MYB recognize a nucleotide sequence motif, PyAACG/TG, that is present in the promoter region of the myb-inducible mim-1 gene and is required for regulation of mim-1 expression by v-MYB and c-MYB. Since the myb binding motif contains a CpG dinucleotide that constitutes a potential target for methylation, we have investigated whether recognition of the binding site by myb proteins is sensitive to CpG methylation of the binding motif. The results presented here demonstrate that bacterially expressed v-MYB as well as authentic v-MYB and c-MYB bind to the myb binding site in a methylation-sensitive manner. Our observations raise the interesting possibility that myb function can be regulated by methylation of myb binding sites. PMID- 8423990 TI - Both the helix-loop-helix and the leucine zipper motifs of c-Myc contribute to its dimerization specificity with Max. AB - The oncoprotein c-Myc contains two dimerization motifs- the helix-loop-helix (HLH) and the leucine zipper (LZ) - through which c-Myc specifically dimerizes with Max. We substituted regions of the c-Myc HLH and LZ motifs with the corresponding regions of structurally related proteins that do not interact with Max. Specific association of c-Myc with Max was dictated by helices 1 and 2 of the HLH motif and by the LZ. Within helix 2, residues 6 and 7 were important determinants of dimerization specificity. c-Myc and Max proteins with three amino acids inserted between their HLH and LZ motifs interacted efficiently, suggesting that the spacing between the motifs can be varied. Furthermore, alignment of the motifs at the primary sequence level was not obligatory, since Max with the three amino acid insertion could interact with wild-type c-Myc. These findings are consistent with our recently proposed model for the architecture of HLH/LZ domains. PMID- 8423991 TI - Negative regulatory elements in the human ETS1 gene promoter. AB - The human ETS1 gene promoter contains binding sites for transcription factors AP1, AP2 and ETS. These factors have been previously shown to be positive regulators for the expression of the ETS1 gene. In our previous report it was inferred that the ETS1 gene promoter also contains negative regulatory elements that might balance and prevent the overexpression of the ETS1 gene product. The studies reported here show that the ETS1 gene promoter also contains binding site motifs for transcription factors PEA3 and OCT. By subjecting deletion constructs of the ETS1 gene promoter to functional analysis, two negative regulatory elements (NREs) were located: NRE1 was mapped to 230 nt at the 5' end of the promoter, and NRE2 was mapped to 350 nt located between the second OCT motif and a 120-base sequence downstream from the SacI site. These NREs can also reduce the activity of a heterogenous promoter. Gel mobility-shift assays showed that nuclear extracts from B-lymphoid (Daudi), T-lymphoid (HPB) and erthyromyeloid (K562) cells each form one major and several minor complexes with NRE1 and NRE2. The nuclear extract from promyelocytic (HL60) cells does not form complexes with either NRE1 or NRE2. The identification of NREs in the ETS1 gene promoter suggests that these elements play an important role in regulating the ETS1 gene expression in hematopoietic cells. PMID- 8423992 TI - Frequent activation of the lck gene by promoter insertion and aberrant splicing in murine leukemia virus-induced rat lymphomas. AB - We have analysed DNA and RNA from 36 T-cell lymphomas induced in Fischer rats by Moloney murine leukemia virus for alterations affecting the structure or expression of the lck gene. At least five primary tumors (14%) have a proviral insertion upstream of lck. In at least four of the tumors, proviral insertion increases lck mRNA levels an average of eight-fold. Overexpression of lck results from transcription initiating in the viral promoter and extending into lck sequences. Three different structures of hybrid transcript were detected. In all three, the hybrid RNAs are spliced to a normal lck splice acceptor in the first exon of lck, resulting in removal of three out of frame ATG codons which would be expected to increase the translation efficiency of the hybrid message. In one tumor, the viral splice donor is used, in one tumor, proviral insertion generates a splice donor sequence one base pair downstream of the long terminal repeat boundary, and in two tumors, a cryptic splice donor in the upstream lck sequences is used. The significance of these unusual splicing patterns and of the higher frequency of proviral insertions adjacent to lck in rats relative to mice is discussed. PMID- 8423993 TI - Ras oncogene transformation of human B lymphoblasts is associated with lymphocyte activation and with a block of differentiation. AB - The p21ras small GTP binding proteins participate in signal transduction from cell surface receptors and affect neoplastic transformation and development in many different cell types. In the present study, we examined the relationship between ras transformation and differentiation of human B lymphocytes. We show that the constitutive expression of the T24 Ha-ras oncogene in EBV-immortalized B lymphoblasts was associated with the induction of the interleukin 2 receptor alpha subunit, with an impaired immunoglobulin gene expression, altered adhesion properties and increased survival in serum-free medium. Since induction of the IL 2 receptor alpha subunit is a hallmark of lymphocyte activation, we suggest that p21ras naturally triggers B cell activation. The ras-transformed lymphocytes displayed a fully functional IL-2r, as assessed by c-fos induction following treatment with IL-2; nevertheless, they were not growth stimulated by this lymphokine. The decreased expression of immunoglobulin genes indicates that the ras oncogene blocks terminal differentiation to plasma cells, possibly by inhibiting the activity of lymphocyte-specific transcription factors. Somewhat unexpectedly, the constitutive p21ras activity did not cause an increased DNA binding of transcription factors PEA1 (AP1), PEA3, Oct-2 or NF-kB. PMID- 8423994 TI - A protein binding site from the murine c-myc promoter contributes to transcriptional block. AB - Recent studies have revealed that expression of several eukaryotic genes can be regulated at the level of transcription elongation. As a first step to elucidate the mechanism by which transcription elongation is modulated, several groups have identified sequences necessary for transcriptional block within the c-myc gene. These studies indicated that transcriptional block depends not only on sequences surrounding the sites of block, but also on sequences within the promoter: some deletions within the c-myc promoter eliminated transcriptional block and, with chimeric constructs, transcriptional block was observed when some heterologous promoters but not others were fused to the c-myc termination region. Using a chimeric construct containing the H-2Kb major histocompatibility class gene promoter linked to the c-myc first exon, we show that transcriptional block is increased by the addition of a 25 bp DNA sequence from the c-myc promoter. Similar results are obtained whether this sequence is inserted upstream or downstream of the transcription initiation site. We further show that nuclear factors interact with this sequence in vitro. Interestingly, when a mutated version of this sequence was tested, we observed decreased nuclear factor binding in vitro as well as reduced transcriptional block in nuclear run-on transcription assays. These results suggest that interactions of protein factors with specific nucleotide sequences near the transcription initiation site can affect elongation of transcription at sites located further downstream. PMID- 8423995 TI - Post-translational modifications of the env-sea oncogene product: the role of proteolytic processing in transformation. AB - The transforming gene product of the S13 avian erythroblastosis virus, env-sea, is a member of the growth factor receptor class of tyrosine kinases. The env-sea precursor protein gp155env-sea is proteolytically processed into the mature cleavage products gp85env-sea and gp70env-sea which are subsequently terminally glycosylated. Previous studies have shown that the abnormal glycosylation of gp155env-sea which takes place in the presence of the inhibitor castanospermine inhibits the proteolytic cleavage of gp155env-sea and blocks its transforming ability. To define a role for proteolytic processing of env-sea in transformation, we have introduced mutations at the protease recognition site which efficiently block cleavage without affecting the biosynthesis or transport of the resulting uncleaved protein. We show here that an uncleaved but fully glycosylated sea-encoded protein retains the ability to transform chicken embryo fibroblasts, indicating that proteolytic processing is not essential for transformation by the env-sea tyrosine kinase. PMID- 8423996 TI - Chromosomal localization of genes encoding the transcription factors, c-rel, NF kappa Bp50, NF-kappa Bp65, and lyt-10 by fluorescence in situ hybridization. AB - We have used fluorescence in situ hybridization (FISH) to perform precise chromosomal mapping of the genes encoding the transcription factors c-rel, NF kappa Bp50, NF-kappa Bp65, and lyt-10. The previously published assignments of c rel and NF-kappa Bp50 have been refined to specific bands. The map position of lyt-10, inferred from its isolation from a t(10;14)(q24;q32) translocation, has been confirmed. NF-kappa Bp65 has now been mapped to 11q13, a site of frequent involvement in aberration in multiple tumor types. PMID- 8423997 TI - Xenopus c-ski contains a novel coiled-coil protein domain, and is maternally expressed during development. AB - v-ski is a virally transduced nuclear oncogene. The biology of its cellular progenitors is only poorly understood. The early development of Xenopus laevis provides an excellent model system for studying the role of genes in proliferation, differentiation and development. We have therefore characterized Xenopus c-ski. We report that Xenopus c-ski transcripts are maternally regulated during early development, and are widely expressed in adult tissues. Structural predictions indicate the presence of a previously unreported extensive C-terminal helical domain in Xenopus c-ski and ski-related proteins from other species. This domain shows homologies with proteins which contain heptad repeats characteristic of elongated coiled-coil formation, like myosin and the intermediate filament proteins, and itself has a heptad repeat structure. The ski C-terminal domain also contains two other previously unreported elements, a novel strictly alternating hydrophobic-basic motif that repeats along the helical domain, and an underlying 25-mer repeated sequence. The significance of these findings is discussed with reference to c-ski function and to the oncogenic activation of v ski, which is a truncated version of c-ski and thus does not have the C-terminal helical domain. PMID- 8423998 TI - Characterization of a positive regulatory element in the mts1 gene. AB - The first intron of the mts1 gene, a gene that is selectively expressed in metastatic cells and in normal cells that are motile, was found to be highly homologous to the CD3 delta enhancer element. Because of the homology between the CD3 delta enhancer and the first intron of mts1, we analysed the first intron of the mts1 gene to determine whether it functions as a transcriptional regulatory element. Highly metastatic CSML-100 cells transfected with chloramphenicol acetyl transferase-containing plasmids demonstrated the ability of the mts1 first intron to function as a positive regulatory element. In vitro footprinting analysis using extracts from CSML-0 cells (which express mts1 at low levels) or CSML-100 cells (which express mts1 at high levels) identified a protected 16-nucleotide element in the first intron of mts1, regardless of the extract used. However, in vivo footprinting analysis of the same region identified the protected 16 nucleotide fragment only in the mts1 intron from CSML-100 cells, not from CSML-0 cells. Differences in the methylation pattern of the mts1 gene in CSML-100 cells and CSML-0 cells are known to exist, and may in part be responsible for the mts1 footprinting differences observed in vivo from the different cell lines. PMID- 8423999 TI - Inducible formation of liver tumors in transgenic mice. AB - Transgenic mice have been generated with an inducible SV40 t/T antigen construct with the aim of analysing the early changes that take place in the course of liver tumorigenesis. The strictly liver-specific human C-reactive protein (CRP) gene promoter was chosen for the control of the transgene expression because this promoter can be turned on transiently by injection of bacterial lipopolysaccharide. Among 10 independently derived CRP-Tag mouse lines five showed inducible expression of the CRP-Tag transgene in liver. However, only one had a tight control of the transgene with virtually no expression under physiological conditions and high levels of Tag expression after stimulation. Females of this line were used to analyse the progression of liver alterations upon repeated induction of the t/T antigen for different lengths of time. The first signs of transgene-induced liver alterations could be monitored by the activation of the marker enzyme gamma-glutamyltranspeptidase 30 days after the start of the induction program. After 90 days hepatocellular carcinomas were already detectable. Thus, CRP-Tag mice constitute an excellent system to analyse the sequential events that take place during liver carcinogenesis. PMID- 8424000 TI - Does television viewing increase obesity and reduce physical activity? Cross sectional and longitudinal analyses among adolescent girls. AB - To examine the relationships between hours of television viewing and adiposity and physical activity among female adolescents, a cohort study with follow-up assessments 7, 14, and 24 months after baseline was conducted. All sixth- and seventh-grade girls (N = 971) attending four northern California middle schools were eligible to participate. Six hundred seventy-one students had sufficient data for baseline cross-sectional analyses, and 279 students in a no-intervention cohort had sufficient data for longitudinal analyses. The baseline sample had a mean age of 12.4 years and was 43% white, 22% Asian, 21% Latino, 6% Pacific Islander, 4% black, 2% American Indian, and 2% other. Hours of after-school television viewing, level of physical activity, and stage of sexual maturation were assessed with self-report instruments. Height, weight, and triceps skinfold thickness were measured and body mass index (ratio of weight [in kilograms] to height [in meters] squared) and triceps skinfold thickness were adjusted by level of sexual maturity for the analyses. Baseline hours of after-school television viewing was not significantly associated with either baseline or longitudinal change in body mass index or triceps skinfold thickness. Baseline hours of after school television viewing was weakly negatively associated with level of physical activity in cross-sectional analyses but not significantly associated with change in level of physical activity over time. All results were essentially unchanged when adjusted for age, race, parent education, and parent fatness. Among adolescent girls, television viewing time appears to have only weak, if any, meaningful associations with adiposity, physical activity, or change in either over time. PMID- 8424001 TI - Effects of television on metabolic rate: potential implications for childhood obesity. AB - The effects of television viewing on resting energy expenditure (metabolic rate) in obese and normal-weight children were studied in a laboratory setting. Subjects were 15 obese children and 16 normal-weight children whose ages ranged from 8 to 12 years. All subjects had two measured of resting energy expenditure obtained while at rest and one measurement of energy expenditure taken while viewing television. Results indicated that metabolic rate during television viewing was significantly lower (mean decrease of 211 kcal extrapolated to a day) than during rest. Obese children tended to have a larger decrease, although this difference was not statistically significant (262 kcal/d vs 167 kcal/d, respectively). It was concluded that television viewing has a fairly profound lowering effect of metabolic rate and may be a mechanism for the relationship between obesity and amount of television viewing. PMID- 8424002 TI - A decade of Medicaid in perspective: what have been the effects on children? AB - This study of the Medicaid program analyzes changes in child recipients, costs, and service use during the 1980s to assess the effects of recent federal policy shifts and to project future costs for children. Data presented in this study are from the Health Care Financing Administration's Medicaid Statistical Report for the years 1979, 1985, and 1990, three time-points that demarcate major federal policy shifts. About half of all recipients added to the Medicaid program during the last decade were children; they comprised 14% of the total cost growth experienced by the program. In addition, the eligibility distribution of children receiving Medicaid shifted markedly over the last decade. In 1979, children receiving cash assistance comprised 90% of total child recipients; by 1990, this figure dropped to 72%. Future expansions to the Medicaid program are projected to cost less than the initial expansions. This is because the early expansions disproportionately served infants, who require more hospital services than older children. Despite the major changes in Medicaid eligibility for children during the 1980s, only limited cost shifts occurred in expenditures for children. Children continue to consume a small portion of the Medicaid budget. Congress should explore options for guaranteeing that their share of funding for services will be adequate. Moreover, since future expansions will be far less expensive than those already implemented, accelerating the phase-in process for all poor children may be a more financially feasible policy option than many policymakers anticipate, despite the fiscal hardships facing many states. PMID- 8424003 TI - Advising parents to stop smoking: pediatricians' and parents' attitudes. AB - Pediatricians are in a unique position to address the issues of smoking cessation with parents. Vermont pediatricians and parents of their patients were surveyed to assess attitudes about giving and receiving smoking cessation advice. A questionnaire was mailed to all pediatricians in Vermont, and 72 valid responses were received, for a response rate of 91%. Forty percent of pediatricians routinely took a smoking history from parents and 11% recorded this information in the child's chart. Most pediatricians (94%) reported advising at least 60% of smoking parents to quit, and they spent an average of 4.4 minutes doing this. Barriers to giving advice were lack of time (42%), feeling that parents did not expect advice (25%), and feeling ill at ease giving the advice (25%). Only 8.5% of pediatricians had received training in how to give smoking cessation advice, but 87% were willing to learn methods to give advice briefly. Six hundred seventy six parents from randomly selected pediatric practices were interviewed. The average parental age was 32, and 84% were women; 49% had never smoked, 30% were former smokers, and 21% were current smokers. Current smokers were less likely to agree with statements about the adverse effects of passive smoke on children. Most parents (56%) felt that pediatricians should give quit-smoking advice to parents, and 52% of smoking parents reported that they would welcome advice. Only 30% of current smokers said advice would bother them somewhat, and 15% had more negative reactions. Parents and pediatricians agreed on the best opportunities to give quit-smoking advice.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424004 TI - Risk factors for delayed immunization in a random sample of 1163 children from Oregon and Washington. AB - Despite extensive study of vaccine safety and decades of effort to immunize infants and toddlers, little is known about the comprehensiveness of vaccine coverage in US children younger than 2 years of age. Provider and parent data from a population-based sample of 1163 children from two states were analyzed to assess coverage rates at three ages and to evaluate characteristics of children and their families that predict failure to immunize on schedule. Overall, 78% of the children had received their first dose of diphtheria and tetanus toxoids with pertussis vaccine (DTP) and their first dose of oral poliovirus (OPV) by 92 days of age. Similarly, 77% had received their third dose of DTP and their second dose of OPV by their first birthday. However, by their second birthday only 60% had received the full series of four doses of DTP, three doses of OPV, and one dose of the measles, mumps, and rubella vaccines. When considered singly, several variables including child birth order, family income, maternal education, and marital status significantly predicted failure to immunize on schedule. In multivariate logistic models, only birth order and maternal education consistently predicted vaccine status at each of the three ages. Compared with first-born children, those who were later-born were 1.7 times more likely to be incompletely immunized at 2 years of age (95% confidence interval: 1.2, 2.3). Children of more educated mothers were significantly less likely to be underimmunized at all ages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424005 TI - Successes and failures in vaccine delivery: evaluation of the immunization delivery system in Puerto Rico. AB - The objective of this study was to evaluate immunization delivery and determine reasons for low coverage among preschool-age public clinic attendees in Puerto Rico. In 25 randomly selected clinics, coverage and missed immunization opportunities were assessed in 273 children aged 2 to 59 months, exist interviews were conducted with parents, and providers were interviewed. Two neighborhoods close to the clinics were surveyed to determine parental knowledge about immunizations, and the vaccination status of children in these neighborhoods was assessed. Two hundred seventy-three clinic attendees were interviewed. Among 229 (84%) with vaccination cards, only 126 (55%) had received all indicated vaccines by completion of the clinic visit. Forty-five percent of children with cards in the household survey were not up-to-date. Of 171 (75%) clinic attendees eligible for vaccination at the visit, 118 (69%) missed one or more immunizations at the visit. In addition, half of all children had previously missed one or more immunizations when they had received another vaccine. Missed opportunities occurred because of nonavailability of vaccines, lack of integration of services, provider misconceptions about contraindications, and failure to administer vaccines simultaneously. Other problems included barriers to immunization services and lack of information and education activities. It is concluded that deficiencies in immunization delivery substantially delay immunization and reduce coverage. PMID- 8424006 TI - The Missouri child fatality study: underreporting of maltreatment fatalities among children younger than five years of age, 1983 through 1986. AB - Estimates of the incidence of child maltreatment fatalities vary widely; most experts believe they are underreported. To investigate the suspicion that fatal maltreatment was underreported in Missouri preschool children, a statewide, population-based study was conducted using nine data sources. The study cases included the 384 children younger than age 5 who died from 1983 through 1986 and whose death certificates were coded with an external cause (injury) or whose deaths were substantiated as abuse or neglect fatalities by the Missouri Division of Family Services. Each fatality was categorized as one of the following: definite maltreatment, probable maltreatment, possible maltreatment, non maltreatment, or inadequate information. Of the 121 cases classified as definite maltreatment, only 47.9% had codes consistent with maltreatment on their death certificates. The Division of Family Services had substantiated 79.3% of definite maltreatment cases as abuse or neglect fatalities. The Federal Bureau of Investigation Uniform Crime Reports database reported only 38.8% of these cases as homicides. In 37.2% of the cases, there was at least one criminal conviction. Child maltreatment fatalities are drastically underreported as such in Missouri because of inadequate investigations, lack of information-sharing between investigators and agencies, and reporting systems that fail to capture the contribution of maltreatment as a cause of death. Missouri has created a statewide system of child fatality review panels and a child fatality surveillance system to address the problems documented in this study. PMID- 8424007 TI - Estimates of fatal child abuse and neglect, United States, 1979 through 1988. AB - The results of recent surveys in the United States have suggested a rising tide of fatalities due to child abuse or neglect (CAN). Because these surveys lack consistency in case definition and are incomplete in coverage, the use of death certificate data to estimate the number of CAN deaths was explored. To estimate these deaths among children 0 through 17 years old for 1979 through 1988, three models were formulated, each comprising six coding categories: (1) deaths coded explicitly as due to CAN, (2) homicides, (3) injury deaths of undetermined intentionality, (4) accidental injury deaths, (5) sudden infant death syndrome fatalities, and (6) natural-cause deaths. Research studies and crime data were relied on to estimate the proportions of deaths in categories 2 through 6 that were actually due to CAN, and other assumptions were varied to create a range of estimates. For the 10-year period, the estimated mean annual CAN fatalities ranged from 861 to 1814 for ages 0 through 4, and from 949 to 2022 for ages 0 through 17. Child abuse and neglect death rates did not increase over the period; in fact, they were relatively stable for ages 0 through 17 and showed a modest decline for 0 through 4. Ninety percent of fatal CAN occurs among children younger than 5 years old, and 41% occurs among infants. About 85% of CAN deaths are recorded as due to other causes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424008 TI - Comparison of childhood burns associated with use of microwave ovens and conventional stoves. AB - To identify the incidence, type, and severity of burns associated with microwave oven (MW) use and to compare MW-associated burns with those associated with use of conventional stoves, we conducted a review of a national data base. Data were obtained from the US Consumer Product Safety Commission Injury Information Clearinghouse for 1986 through 1990 concerning burn injuries to children (0 to 19 years). There were an estimated 5160 burns associated with MW use. The mean age was 7.6 years (median, 6 years); 25% of burns were to children younger than 36 months old. Fifty-eight percent involved females. Most MW burns were scalds (95%); 16% of these scalds were from exploding eggs or other food. No MW burn involved a body surface area greater than 25% and no patient required hospital admission. Microwave oven burns were compared with stove burns. There were an estimated 41198 stove-associated burns to children. The mean age was 5.8 years; the median was 3 years. Forty-five percent of burns were to children younger than 36 months old; 55% were to males. Most stove burns (74%) were thermal; 7% involved a body surface area greater than 25%. Five percent of children with stove burns required hospital admission. We conclude that (1) burns to children associated with MW use are less frequent and less severe than stove burns; (2) MW burns predominantly affect females; and (3) burn prevention efforts should emphasize the hazards of stoves, which vastly exceed those of MWs. PMID- 8424009 TI - Cocaine use in women from a defined population: prevalence at delivery and effects on growth in infants. AB - Adverse health effects associated with intrauterine cocaine exposure (prematurity and its associated morbidity, intrauterine growth retardation, possible risk of sudden infant death syndrome) are based on studies from large urban hospitals, but few data exist from other sources. The current study, set in a community hospital, was designed to (1) estimate the prevalence of maternal cocaine use at delivery, (2) describe neonatal outcomes, and (3) evaluate physiological growth in exposed children. The study was conducted over 30 months (total births were 14,074) at The Children's Hospital of Greenville Memorial Hospital, the major source of neonatal care for Greenville County, South Carolina (1990 population: 320,000). A child was considered exposed to cocaine if there was documented evidence of use in the mother's medical record or if one member of the pair had a positive urine drug screen. Growth data were abstracted from clinical records. Overall prevalence of exposure was 1.0%. Of the 137 subjects (89, positive urine drug screen; 48 self-reported exposure), 21 (15%, 95% confidence interval, 9% to 21%) were premature (gestational age < 37 weeks) and 2 died of sudden infant death syndrome. Mean age- and sex-adjusted percentiles for weight, length, and head circumference increased from 23%, 29%, and 18%, respectively, at birth to 43%, 49%, and 54% in children followed for 12 months; however, 50% of the cohort were lost to follow-up, and these children were smaller at birth than those under active follow-up. Rates of prematurity and infant death were similar to those reported in urban hospitals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424010 TI - Congenital conductive hearing loss: the need for early identification and intervention. AB - Pediatricians are familiar with conductive hearing loss, but to many it is synonymous with otitis media of some type. There is a group of children who have anatomic abnormalities of the external or middle ear causing their hearing losses. The records of 565 hearing-impaired children were reviewed, and 54 with non-otitis-related conductive hearing loss were found. Date of birth, degree of loss, diagnosis or presence of any associated abnormalities, and ages of confirmation and intervention were studied. There were 22 children with microtia, 14 with abnormalities of the external auditory canal and normal pinnae, 15 with obvious dysmorphic features including 9 who had syndromes well known to be associated with hearing abnormalities, and only 3 who had isolated hearing loss. Despite all these clues, hearing loss was diagnosed in only 10 children (excluding those with bilateral microtia) before 12 months of age although their average hearing loss was 45 dB. It is important for the primary care provider to recognize these children as early as possible as their loss is permanent and almost always more severe than that seen in otitis. PMID- 8424011 TI - Evaluation of pediatric training by the alumni of a residency program. AB - An evaluation of a large pediatric residency program by its alumni was undertaken to determine whether the program is providing quality education and relevant training for pediatricians. A questionnaire was mailed to physicians who began their training at Children's Hospital Medical Center in July 1974 or later and finished prior to July 1990. Of the 372 questionnaires sent, 274 were returned (74% response rate). Current positions included primary care physicians (41%), academic pediatricians (33%), and other (26%). Fifty-seven percent pursued fellowship training. Overall, 94% of the responders believed that their pediatric training was adequate in preparing them for their current position. The variety of patients seen, the inpatient experience, resident camaraderie, and neonatology training were rated the highest quality, means 4.38 to 4.75 (1 = poor, 3 = average, 5 = excellent quality). Behavioral/developmental pediatrics, adolescent medicine, well-child care/continuity clinic, the outpatient experience, and pediatric surgery were rated the poorest quality, means 2.66 to 3.08, and judged to have too little quantity of training relative to the other aspects of training, means 1.93 to 2.42 (1 = too little, 3 = just enough, 5 = too much quantity). The quantity of neonatology training, the volume of patients, on-call experience, subspecialty inpatient services, such as bone marrow transplantation, and overall inpatient experience were considered too high relative to other components, means 3.51 to 4.08. There were no significant differences in responses by whether the alumni were in academic or primary care positions nor by year of training, and few differences by whether or not the training was considered adequate preparation for their current position.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424012 TI - Cost assessment of the evaluation of heart murmurs in children. AB - Echocardiography available directly to primary care physicians represents an alternative strategy to pediatric cardiology consultation for childhood heart murmur evaluation. A decision analysis model compared the costs of two diagnostic strategies: (1) echocardiography first, referral to the cardiologist if appropriate; and (2) cardiologist evaluates murmur, echocardiography if appropriate. The model incorporated probability of heart disease, frequency of cardiologist-ordered echocardiography, and echocardiography results established by review of 388 records of consecutive patients evaluated for murmurs in a pediatric cardiology clinic. Echocardiography-first strategy costs were $257 greater than referral-strategy costs. Referral-strategy advantages were not highly sensitive to changes in model assumptions. Pediatric cardiology consultation is the preferred approach provided (1) consultation costs are moderate, (2) echocardiography costs are moderate to high, and (3) the rate at which the cardiologist orders echocardiography for patients with innocent murmurs is low. PMID- 8424013 TI - Pediatric intensive care sedation: survey of fellowship training programs. AB - Children hospitalized in a pediatric intensive care unit are frequently distressed. The purpose of this study was to identify the patterns of use of sedative agents in pediatric critical care patients. A questionnaire survey was mailed to 45 directors of Pediatric Critical Care Fellowship Training Programs listed in Critical Care Medicine, January 1989. The response rate was 75.6% (34 questionnaires). The most commonly identified goals of sedation were reduced patient discomfort or distress and fewer unplanned extubations. The agents most frequently employed for this purpose were opioids (morphine or fentanyl), chloral hydrate, or benzodiazepines. Although conventional doses are used, opioids and benzodiazepines are often given hourly or by continuous infusion. Satisfaction with the efficacy and safety of commonly used opioids was greater (most common response "very satisfied") than for the benzodiazepines ("somewhat satisfied"). The physician's or nurse's clinical impression was reported to be the "most important" criterion for deciding when a patient required a dose of sedative; objective criteria were selected as less important. The majority of patients (65.7%) in the surveyed units were ideally "sedated to the point of no distress with as-needed medication." The majority of respondents (76.4%) identified efficacy as the major problem with sedation. Drug withdrawal was considered to be the major problem with sedative use by only a minority of respondents (6.9%). Although withdrawal is seen in 61.8% of units, it is generally treated when recognized, rather than prevented by routine tapering of sedation. Optimal sedation of pediatric intensive care unit patients is considered problematic, despite the use of frequent doses of many sedatives. Systematic investigation of pharmacodynamic response to these agents in the pediatric critical care population is indicated. PMID- 8424014 TI - Role of local anesthesia during lumbar puncture in neonates. AB - Local anesthesia decreases physiologic responses to pain in neonates but has not been used routinely during lumbar punctures in newborns, as it might obscure anatomical landmarks. However, local anesthesia may decrease newborns' struggling during lumbar puncture, thus facilitating the procedure and increasing its success rate. The success rate of lumbar punctures was compared in neonates allocated prospectively to 0.2 to 0.5 mL of 1% lidocaine anesthesia (n = 48) or a control group (n = 52). Newborns were held in a modified lateral recumbent position (neck not flexed) and their struggling response to the various steps in the lumbar puncture was scored by the holder. The newborns' struggling motion score increased in response to lidocaine injection, but response to the subsequent spinal needle insertion was significantly decreased. Despite this decreased motion, no differences were noted in the number of attempts per lumbar puncture (1.9 +/- 0.2 [SEM] in lidocaine and 2.1 +/- 0.2 in control groups), rate of lumbar puncture failure (15% in lidocaine and 19% in control groups), or the number of traumatic lumbar punctures (46% in both groups). The success rate of lumbar puncture was not dependent on level of training of physicians performing the procedure. No acute complications, cerebrospinal fluid contamination, or subsequent meningitis was noted in either group. It is concluded that local anesthesia with lidocaine decreases the degree of struggling but does not alter the success rate of lumbar puncture in neonates. The practice of withholding lidocaine anesthesia from neonates undergoing lumbar punctures cannot be justified by arguing that it makes the procedure more difficult to perform. PMID- 8424015 TI - Family correlates of a 10-year pulmonary health trend in cystic fibrosis. AB - In this study, 33% of the variance in the 10-year trend of forced expiratory volume in 1 second (FEV1), used as an index of pulmonary health, was explained by differences in family characteristics at the start of the study. Balanced family coping, a family emphasis on personal growth, and compliance with treatment for 91 children with cystic fibrosis were assessed at the start of the study and the FEV1 was observed at every clinic visit over the next 10 years. When both parents' coping emphasized family integration, support for self, and medical consultation, the FEV1 trend was better. Compliance with daily chest physical therapy and with quarterly clinic visits was associated with a better FEV1 trend. Poorer FEV1 trend was associated with active social involvement of family members. Older patients and patients whose parents worked more hours outside the home had lower compliance. These findings support the importance of encouraging families to balance their resources between the child's health needs and family needs. PMID- 8424016 TI - Perianal injuries resulting from sexual abuse: a longitudinal study. AB - Four children who incurred perianal injuries as a result of a sexual assault were followed on a longitudinal basis to document the anatomical changes that ensued. The subjects, whose ages ranged from 4 to 8 years, were followed from 1 week to 14 months. They were examined in both supine and prone knee-chest positions and a 35-mm camera mounted on a colposcope was used to record their injuries. At the time of the initial examination, there were a variety of findings including erythema of the tissues, edema of the skin folds, localized venous engorgement, dilation of the external anal sphincter, and lacerations of different depths. Superficial lacerations reepithelized within 1 to 11 days. The second-degree wounds in two of the children were healed by the 1- and 5-week return visits, leaving narrow bands of scar tissue. In the two subjects who were followed the longest, signs of both a second-degree laceration and a surgically repaired third degree injury had virtually disappeared by 12 to 14 months after the assaults. The wounds in one subject, infected with a herpes simplex type 2 virus, remained erythematous for a longer period of time than did similar injuries in the other children. A skin tag created by the avulsion of the tissues in one subject persisted, although it became less obvious as it retracted into the redundant folds of the perianal tissues over time. PMID- 8424017 TI - Prader-Willi syndrome: consensus diagnostic criteria. AB - The diagnosis of Prader-Willi syndrome (PWS) is based on clinical findings that change with age. Hypotonia is prominent in infancy. Obesity, mild mental retardation or learning disability, and behavior problems, especially in association with food and eating, result in a debilitating physical and developmental disability in adolescence and adulthood. No consistent biological marker is yet available for PWS in spite of recent research activity in cytogenetics and molecular genetics. Diagnostic criteria for PWS were developed by consensus of seven clinicians experienced with the syndrome in consultation with national and international experts. Two scoring systems are provided: one for children aged 0 to 36 months and another one for children aged 3 years to adults. These criteria will aid in recognition of the syndrome in hypotonic infants and in obese, mildly retarded, behaviorally disturbed adolescents and adults. They will also ensure uniform diagnosis for future clinical and laboratory research in PWS. PMID- 8424018 TI - Common pulmonary vein atresia: the role of extracorporeal membrane oxygenation. AB - Common pulmonary vein atresia is a rare form of cyanotic congenital heart disease in which the pulmonary veins join to form a blind confluence that does not communicate with the heart or the major systemic veins. Twenty-one cases have been reported since the lesion was first described in 1962; only two patients with this lesion have survived. Over a 4-year period, common pulmonary vein atresia was diagnosed in five newborns referred to the San Diego Regional Extracorporeal Membrane Oxygenation Program. All five improved dramatically as a result of venoarterial bypass. Congenital heart disease was diagnosed at autopsy in the initial case and by cardiac ultrasound and/or catheterization in the others. Surgical repair was attempted in three neonates; all three required continued extracorporeal membrane oxygenation support postoperatively because of pulmonary hypertension and severe pulmonary parenchymal disease. One infant died of respiratory insufficiency at 3 months of age. The other two survived and were discharged from the hospital. The diagnostic and therapeutic dilemmas posed by this lesion and the life-saving potential for extracorporeal membrane oxygenation in this rapidly fatal cardiac anomaly are the bases of this report. PMID- 8424019 TI - Extent of duplication in lower-limb malformations suggests the time of the teratogenic insult. AB - Investigations of vertebrate limb development have suggested that a process called "specification" instructs the cells of the future limb as to which tissues they should form. This process proceeds in a wave-like manner, starting at the most proximal levels of the future limb and ending at its distal tip. Human limb specification probably occurs during the fourth and fifth weeks of development. It is proposed that human limb duplications result from errors of specification and, furthermore, that the more distal the duplication, the later the occurrence of the teratogenic event during the specification process. Therefore, among human lower limbs with duplications, one may be able to estimate the relative time of the teratogenic event by comparing the levels at which the duplications occur. PMID- 8424020 TI - Report of the National Institute of Neurological Disorders and Stroke workshop on near infrared spectroscopy. PMID- 8424021 TI - Severe acquired protein C deficiency in purpura fulminans associated with disseminated intravascular coagulation: treatment with protein C concentrate. PMID- 8424022 TI - Fatal child abuse and sudden infant death syndrome: a critical diagnostic decision. PMID- 8424023 TI - Adolescent health status measurement: development of the Child Health and Illness Profile. AB - This report describes the early stages in the development and testing of an instrument, known as the CHIP (Child Health and Illness Profile), for assessing the health of individuals aged 11 through 17. The purpose of the instrument is to assess health in epidemiologic surveys, to determine the existence of systematic differences in health in subpopulations (including the socioeconomically disadvantaged), and to provide a basis for assessing the impact of changes in health services or health policies. An instrument consisting of six domains with 25 subdomains was developed based on the literature, the involvement of focus groups and expert panels, and pretesting in four groups of teenagers known to differ in their health. The results of work with panels of experts suggest that the instrument has content validity. Most domains and subdomains had acceptable reliability as measured by alpha coefficients. Differences in the scores of individuals in the four groups were in the predicted directions, suggesting that the instrument also has construct validity. Additional research is under way to establish other aspects of validity as well as reliability in school populations of adolescents as well as specific clinical settings. PMID- 8424024 TI - Help me make it through the night: behavioral entrainment of breast-fed infants' sleep patterns. AB - The study objective was to investigate whether exclusively breast-fed infants could be taught to sleep through the night (defined from 12:00 AM to 5:00 AM) during the first 8 weeks of life. The design was short-term longitudinal, from the last trimester of pregnancy until the eighth week after birth. Twenty-six first-time parents and their newborn were randomly assigned to treatment and control groups (13 in each group). Treatment parents were instructed to offer a "focal feed" (between 10 PM and 12 AM) to their infants every night, to gradually lengthen intervals between middle-of-the-night feeds by carrying out alternative caretaking behaviors (eg, reswaddling, diapering, walking), and to maximize environmental differences between day and nighttime. All parents kept 72-hour diaries of their infants' feeding and sleeping patterns every week from birth to 8 weeks of age and rated their infants' temperament at birth and at 8 weeks. By 3 weeks, treatment infants showed significantly longer sleep episodes at night. By 8 weeks 100% of treatment infants were sleeping through the night compared to 23% of control infants. Treatment infants were feeding less frequently at night but compensated for the relatively long nighttime interval without a feed by consuming more milk in the early morning. Milk intake for 24-hour periods did not differ between groups. Treatment infants were rated as more predictable on Bates' Infant Characteristics Questionnaire. It is concluded that parents can have a powerful influence on the development of their infants' sleep patterns. Frequent night waking in breast-fed infants often results in early termination of lactation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424025 TI - Epidemiology of Rett syndrome: a population-based registry. AB - The Texas Rett Syndrome Registry maintains the largest population-based registry of cases and potential cases of Rett syndrome in the world. The most precise estimate of the prevalence of Rett syndrome of 1 per 22800 (0.44/10000) females aged 2 through 18 years of age was generated from this Registry. In addition, the first prevalence figures for black and Hispanic female cases were estimated. Registry cases are actively ascertained from multiple sources. Registry staff identify presumptive cases from review of information provided to the Registry by the parent or guardian. Preliminary diagnostic evaluation includes standardized review of medical records and videotape of key behaviors. Diagnosis is confirmed at clinical evaluation. The active surveillance system is monitored with the two source capture-recapture methodology and case ascertainment is projected. The 1990 prevalence estimate of Rett syndrome indicates that the syndrome occurs less frequently than previously estimated. Until a biologic marker for Rett syndrome is identified or a standard definition for an incident case of Rett syndrome is designated, the prevalence of Rett syndrome will remain a major investigative issue of its epidemiology, and the Registry will be an important, systematic mean to gather case material for clinical and laboratory studies providing the foundation for the development of preventive interventions. PMID- 8424026 TI - Serum bactericidal test as a prognostic indicator in acute pulmonary exacerbations of cystic fibrosis. AB - The serum bactericidal test has been used for many years for optimal assessment of the efficacy of antibiotic therapy in patients with infective endocarditis and other bacterial infections. Its capacity to predict the bacteriological outcome of acute pulmonary exacerbations in patients with cystic fibrosis was evaluated. A total of 54 courses of intravenous antibiotic therapy were analyzed in 22 patients, whose ages ranged from 4 months to 24 years (mean age: 10 years). The serum bactericidal activity of blood samples, taken at expected peak and trough antibiotic levels on day 4 of therapy, were determined against the potentially pathogenic strains isolated in sputum at the time of admission. For 104 isolates (64 Pseudomonas aeruginosa, 28 Staphylococcus aureus, and 12 Haemophilus influenzae strains), the peak and trough bactericidal titers were compared to bacteriological outcome. Bacteriological success was defined as a decrease of 2 log10 units or more in the bacterial density in sputum between days 0 and 7 of therapy. At peak antibiotic levels, serum bactericidal titers of 1:128 or more were 96% (all isolates) and 89% (P aeruginosa isolates), predictive of cure, whereas serum bactericidal titers of less than 1:16 were 100% predictive of failure for all infecting bacteria. In patients aged less than 18 years, the best peak titer for predicting success was 1:64, with a predictive value of 96% for titers of 1:64 or greater. The peak titer that best predicted success in patients aged 18 years or more was 1:128, with a predictive value of only 83% for titers of 1:128 or greater.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424027 TI - Early Lyme disease: a flu-like illness without erythema migrans. AB - The existence of a form of early Lyme disease characterized by a flu-like illness without erythema migrans is controversial. To confirm the existence and define the clinical characteristics of the flu-like illness without erythema migrans of localized Lyme disease, the authors studied patients from a Lyme disease endemic area of Connecticut who visited their primary care physicians with an undefined flu-like illness. Patients kept a diary of their symptoms. Acute and convalescent sera were obtained. The diagnosis of Lyme disease was based on the appearance of IgM or IgG antibodies to Borrelia burgdorferi as demonstrated by both enzyme linked immunosorbent assay and immunoblot assay. Twenty-four untreated patients were studied. In five patients acute serologic evidence of Lyme disease developed. The flu-like illness in these five patients was characterized by fever and fatigue and resolved spontaneously in 5 to 21 days. Symptoms recurred in three of these five patients. The existence of a flu-like illness without erythema migrans of early Lyme disease has been clearly established. Prospective, controlled studies are needed to better define its incidence, characteristics, and prognosis so that appropriate diagnostic and therapeutic strategies can be developed. PMID- 8424028 TI - Neonatal intensive care and birth weight-specific perinatal mortality in Michigan and Lorraine. AB - This study investigated the factors influencing use of neonatal intensive care and perinatal mortality in regions of the United States and France, two countries with similar health care systems but different approaches to health financing. The study employed birth certificates from Michigan and a birth registry from Lorraine in 1984. The study showed that geographic access and socioeconomic status were important in determining use of neonatal intensive care in both regions. Socioeconomic factors in perinatal mortality were also shown for both regions, after controlling for gestational age, birth weight, and neonatal intensive care use. In Michigan, infants of mothers with low education had higher mortality rates and in Lorraine residents of low income areas had higher mortality rates. A higher proportion of Michigan women delivered in hospitals with neonatal intensive care than in Lorraine, in all weight/gestation categories. Perinatal mortality rates were also lower in Michigan than in Lorraine, overall and within birth weight categories. PMID- 8424029 TI - Hyperbilirubinemia in the breast-fed newborn: a controlled trial of four interventions. AB - A controlled clinical trial was conducted to compare the effect of four different interventions on hyperbilirubinemia in 125 full-term breast-fed infants. Of 1685 term infants who met the inclusion criteria, 126 (7.4%) had a serum bilirubin concentration > or = 291 mumol/L (17 mg/dL). When the bilirubin reached this level, babies were assigned at random to one of four interventions: (1) continue breast-feeding and observe; (2) discontinue breast-feeding, substitute formula; (3) discontinue breast-feeding, substitute formula and administer phototherapy; (4) continue breast-feeding, administer phototherapy. The serum bilirubin concentration reached 342 mumol/L (20 mg/dL) in 24% of infants in group 1, 19% in group 2, 3% in group 3, and 14% in group 4. When phototherapy was used, the decline in serum bilirubin was significantly larger and more rapid (compared with no phototherapy). In the majority of breast-fed infants whose serum bilirubin levels reach 291 mumol/L (17 mg/dL) the bilirubin will decline spontaneously and will not reach 342 mumol/L (20 mg/dL). If the infant is significantly jaundiced and a decision is made to intervene, parents can be given a number of options and can make an informed decision regarding which, if any, intervention they prefer. PMID- 8424030 TI - Relationship between periventricular intraparenchymal echodensities and germinal matrix-intraventricular hemorrhage in the very low birth weight neonate. AB - The pathogenesis of the periventricular intraparenchymal echodense lesion (IPE) observed in association with germinal matrix-intraventricular hemorrhage (GM-IVH) in premature neonates is unclear. The objectives of this study were to determine (1) the temporal characteristics of GM-IVH and IPE, (2) the basic characteristics of the IPE, and (3) the relationship of clinical events, including surfactant administration, to IPE. One hundred twenty-four neonates of less than 1250 g birth weight were prospectively evaluated. IPE was defined as an echodensity greater than 1 cm in diameter by cranial sonography. Fifteen (12%) neonates developed IPE in association with GM-IVH (group 1); 33 neonates developed GM-IVH only (group 2) and 76 neonates without GM-IVH served as comparison group (group 3). IPE was essentially an asymmetrical lesion; both sides of cerebrum were equally affected. The lesion was diffuse in 9 neonates and focal in 5. IPE occurred both early, at 36 hours or before (n = 8), and later, ie, between 48 and 96 hours (n = 6). In one neonate IPE was diagnosed at autopsy. GM-IVH and IPE were noted simultaneously in neonate with the earlier onset IPE (diagnosed within 36 hours); GM-IVH preceded the IPE by 6 to 48 hours when the lesion was of a later onset. Surfactant was administered to 13 (87%) group 1, 24 (73%) group 2, and 35 (46%) group 3 neonates. Pulmonary hemorrhage developed in 9 (60%) of group 1, 3 (9%) group 2, and no group 3 neonates. Symptomatic patent ductus arteriosus occurred in 12 (75%) group 1, 15 (45%) group 2, and 15 (20%) group 3 neonates. The onset of symptoms associated with patent ductus arteriosus was earlier in group 1 vs group 2 or group 3 neonates, ie, 70 vs 172 hours. Nine (60%) group 1 neonates, 6 (18%) group 2, and 5 (7%) group 3 neonates died. The cranial sonogram was markedly abnormal in all 6 group 1 survivors. Stepwise polytomous logistic regression indicated that birth weight, gestational age, and emergent cesarean section were the best predictors of GM-IVH + IPE.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424031 TI - Injuries in children of adolescent mothers: home safety education associated with decreased injury risk. AB - The purpose of this study was to describe the epidemiology of injuries in children of adolescent mothers (< 18 years old at delivery), in Baltimore, MD, and to explore the relationship between maternal receipt of home safety information and child injury. A random sample of 363 adolescent mothers and their children were followed longitudinally by home interview at 3 and 15 months postpartum. Receipt of home safety information and information source were assessed at the 3-month interview. Injuries requiring medical attention were assessed at the 15-month interview. Sixty-eight children sustained injuries during follow-up and 14% required hospitalization. Falls and burns predominated as the cause of injury, with burns much more common in girls. The children of mothers who received home safety information from family and community-based sources by 3 months postpartum had significantly lower risk of injury during follow-up than children of mothers who had not received home safety information. As the number of information sources increased, the injury rate decreased. Further work is needed to examine the most appropriate timing, repetition, format, and content of injury prevention education. PMID- 8424032 TI - Wisconsin cystic fibrosis chest radiograph scoring system. AB - A new clinical scoring system for patients with cystic fibrosis is needed because of recent advances in diagnosis and treatment which have changed the course of this disease. Chest radiograph scoring is the best objective measure of pulmonary disease for longitudinal studies beginning with infants; however, based on pilot studies, previous scoring systems are not sensitive enough in discriminating between degrees of mild lung disease. Therefore, a new radiographic scoring system was developed with the goal of achieving both sensitivity and reproducibility. This objective was pursued by applying multiattribute utility theory, using a panel of interpreters with expertise in cystic fibrosis radiology, and employing mathematical modeling techniques to weight the various components. The system was developed and validated in three phases including comparison to the Brasfield method of quantitative radiology. The data demonstrate that the new system can be applied reliably and conveniently to generate reproducible scores of pulmonary disease severity. Evaluation of the scores by four independent raters using chest radiographs from 61 patients at an average age of 8.37 years revealed good agreement with a .714 Kendall coefficient of concordance. Assessment of serial changes over time was performed using a group of 176 chest radiographs from 25 patients ranging from 4 weeks to 6 years old; this showed that the Wisconsin system generates score differences that are greater in magnitude with disease progression compared with the Brasfield method. Therefore, the new method is more sensitive to progression of mild disease and should be superior to prior radiographic scoring systems for evaluating therapies designed to modify the early course of disease. The Wisconsin system is designed to be useful in longitudinal clinical studies involving young children with cystic fibrosis and is capable to detecting progression from normality to mild lung disease. PMID- 8424033 TI - Concerning the American Academy of Pediatrics recommendation on sleep position for infants. PMID- 8424034 TI - TV or not TV: fat is the question. PMID- 8424035 TI - Effects of technologic and sociocultural changes on the practice of neonatal medicine: a view from France. PMID- 8424036 TI - American Academy of Pediatrics Committee on Child Health Financing: Principles of child health care financing. PMID- 8424037 TI - American Academy of Pediatrics Committee on Child Health Financing: Scope of health care benefits for infants, children, and adolescents through age 21 years. PMID- 8424038 TI - Acrodermatitis without evidence of bowel disease. PMID- 8424039 TI - Listening to parents. PMID- 8424040 TI - SIDS and sleeping position. PMID- 8424041 TI - The changing role and responsibilities of a chairman. PMID- 8424042 TI - Exercise and breast-feeding: a different experience. PMID- 8424043 TI - The use of cow's milk in infancy. PMID- 8424044 TI - Anaphylaxis at school: etiologic factors, prevalence, and treatment. PMID- 8424045 TI - How much resuscitation is enough resuscitation? PMID- 8424046 TI - Inappropriate statistics. PMID- 8424047 TI - Discharge milk samples. PMID- 8424048 TI - A reply to Scarr and Ernhart. PMID- 8424049 TI - [Biology of aging blood vessels]. AB - The localisation of arteriosclerosis to foci of haemodynamic stress of sites of supporting collagen or elastic fibre deficiency illustrates the importance of mechanical factors in the fate of the smooth muscles. Excessive stress depletes stocks of ATP (adenosine triphosphate), resulting in osmotic lysis. Phosphate ions are released and precipitated in calcifications as apatite. The vascular lesion is followed by reparative and adaptive tissue inflammation. The haemodynamic stress is modified by a number of psychosocial factors, and is increased by traumatic and other injury to the ground substance. Some of the LDL (low-density lipoprotein) perfusing the intima is captured in the proteoglycan matrix, especially if the media is thickened. When cells and ground substance are lysed, fractions are incorporated in macrophages through metabolically uncontrolled receptors. Toxic oxidised LDL is then continuously ingested in the cells before they eventually die, leaving the amorphous lipid-rich mass of which an atheroma is formed. The macrophages produce free radicals which accelerate the damage and the inflammation. Thus, many factors working at different levels are involved in the aetiology of arteriosclerosis, with or without atheromatosis. The only theoretically feasible though insufficiently tested dietary recommendation for the general population would be to reduce the in vivo redox potential of food intake. PMID- 8424050 TI - [International Alzheimer research--from management to molecular biology]. PMID- 8424051 TI - [Scandinavian physicians and European cooperation in 1993]. PMID- 8424052 TI - [Positively-charged health campaign in Finland: the healthy brain]. PMID- 8424053 TI - [Scandinavian Summer school in cancer epidemiology]. PMID- 8424054 TI - [EEC and the Scandinavian labor market agreement]. PMID- 8424055 TI - [Consequences of the EEC agreement for physicians]. PMID- 8424056 TI - [Medical specialties in the EEC region]. PMID- 8424057 TI - [Physicians' organizations in Europe]. PMID- 8424058 TI - [Drug questions in the EEC region]. PMID- 8424059 TI - What's new in breast cancer? Part II. PMID- 8424060 TI - [The Scandinavia Jahre Prize]. PMID- 8424061 TI - [Grand Jahre Prize 1992. functional architecture of adrenal cortex hormone receptors]. AB - The Jahre Prize in medicine (or the 'Little Nobel', as it is also called) for 1992 has been awarded to Professor Jan-Ake Gustafsson, Stockholm, for his pioneering research on the regulation of genetic capacity by steroid hormones. His award lecture, The functional architecture of corticosteroid receptors, consisted in a survey of available knowledge of the structure of corticosteroid receptors and their mechanism of action. PMID- 8424062 TI - Effects of communication training on parents and young adolescents. AB - Two hundred fifty-eight middle school children and their parents attended communication skills training sessions for 6 weeks. Measures of satisfaction with the family system and communication of this group were compared with those of 201 young adolescents and parents who did not attend the sessions. Young adolescents, mothers, and fathers who were trained in communication skills reported increased satisfaction with the family system and open communication immediately and 6 months after training. Young adolescents who were not trained became increasingly dissatisfied with the family system. A subsample of families were observed resolving conflicts. Fathers and young adolescents who were in training used less antisocial and negative problem-solving techniques than those who were not trained. The authors cautiously recommend communication skill training as a "rite of passage" for families with young adolescents. PMID- 8424063 TI - Deciding on breast cancer treatment: a description of decision behavior. AB - The purposes of this study were to describe the unaided decision-making process of women facing treatment for early stage breast cancer and to provide the empirical grounding to develop a conceptual framework for more structured research. A convenience sample of 48 women completed an open-ended interview while they were making a decision. A qualitative analysis of these interviews identified five empirical indicators of decision behavior: (a) perceived salience of alternatives, (b) decision conflict, (c) information seeking, (d) risk awareness, and (e) deliberation. The configurations of these indicators discriminated the decision maker into one of three empirically derived groups. In order of complexity, these are termed Deferrer, Delayer, and Deliberator. PMID- 8424064 TI - Replication and validation of anticipated turnover model for urban registered nurses. AB - Replication of the Anticipated Turnover Model Among Nurses for urban registered nurses provided substantial support for the stability and generalizability of the theoretical model. In both the original and replication studies, causal modeling was used to test the theoretical model predicting job satisfaction, anticipated turnover, and actual turnover. The replication study included 385 full-time nurses in two public and two private urban hospitals. The replication validated, with younger, more educated staff, the major findings from the original study. Group cohesion and job satisfaction effectively predicted anticipated turnover in the replication and the original study. Anticipated turnover was a good predictor of actual turnover, with discriminant analysis yielding 73.2% successful predictions in the replication study and 76.2% in the original study. Job satisfaction effectively buffered job stress. The replication also substantiated the position that job satisfaction strategies need to be targeted specifically to the types of clinical services. PMID- 8424065 TI - How to get to the point. PMID- 8424067 TI - Teetering on the edge: a substantive theory of postpartum depression. AB - Using the grounded theory method, a substantive theory of postpartum depression was developed. Data were obtained through participant observation in a postpartum depression support group over an 18-month period. In addition, 12 in-depth taped interviews were conducted with mothers who had attended the support group. Analysis of the data by the constant comparative method revealed loss of control as the basic social psychological problem. Women suffering from postpartum depression attempted to cope with the problem of loss of control through the four stage process of teetering on the edge. The stages that emerged from the data included (a) encountering terror, (b) dying of self, (c) struggling to survive, and (d) regaining control. PMID- 8424066 TI - Nurses' job satisfaction: a meta-analysis of related variables. AB - The purpose of this study was to describe the magnitude of the relationships between nurses' job satisfaction and the variables most frequently associated with it. A meta-analysis of data from 48 studies with a total of 15,048 subjects revealed that job satisfaction was most strongly associated with stress (-.609) and organizational commitment (.526). Seven variables had correlations between .20 and .50: communication with supervisor, autonomy, recognition, routinization, communication with peers, fairness, and locus of control. Four other variables frequently included in these studies had low correlations (less than .20): age, education, tenure, and professionalization. The influence of employment site, date of study, and measures used on the size and consistency of estimates was described. PMID- 8424068 TI - Effects of information on adaptation to cesarean birth. AB - The effects of cesarean birth information given in childbirth preparation classes on maternal postpartum reactions to unplanned cesarean delivery were examined. An experimental group received comprehensive cesarean birth information as part of standard childbirth preparation classes. The control group received a standard childbirth preparation class curriculum that included limited cesarean birth information. The control group reported a slight decline in pain intensity from 1 2 days to 6 weeks postpartum, whereas the experimental group showed no change over time. No differences between the two groups were found in perception of the birth experience, physical distress, self-esteem, functional status, feelings about the baby, or quality of the marital relationship. The lack of substantial differences between the groups may be due to the normalizing effect of the high cesarean birth rate and greater attention given to this method of childbirth by expectant parents and childbirth educators. PMID- 8424069 TI - Assessment of postoperation pain in children and adolescents using the adolescent pediatric pain tool. AB - The location, intensity, and quality of pediatric postoperative pain were assessed in a convenience sample of 65 multiethnic children and adolescents, 8 to 17 years old. Pain was measured daily for 5 days during hospitalization using the Adolescent Pediatric Pain Tool (APPT). Mean pain intensity scores and mean number of pain descriptors (quality) decreased over time, but there was no significant change over time for the number of body segments marked (location). The findings provided valid and reliable estimates of adolescents' and children's self-reports of the location, intensity, and quality of postoperative pain. PMID- 8424070 TI - Guidance for managing a research grant. PMID- 8424071 TI - Once the dye is cast. Interview by Catharine Sadler. PMID- 8424072 TI - Taking liberties? PMID- 8424073 TI - Crucial choices. PMID- 8424075 TI - Charter of charters? PMID- 8424074 TI - Everybody's challenge. PMID- 8424076 TI - Women and alcohol. Under the influence. PMID- 8424077 TI - Women and alcohol. Tackling dependency. PMID- 8424078 TI - The wrong way to help. PMID- 8424079 TI - Learning cues. PMID- 8424080 TI - Catheter concerns. PMID- 8424081 TI - Nutrition in pregnancy. PMID- 8424082 TI - Speaking up. PMID- 8424083 TI - Baptism of fire. Interview by Daloni Carlisle. PMID- 8424084 TI - Systems of life. The nervous system. 4. PMID- 8424085 TI - Profession mourns loss of AIDS pioneer. PMID- 8424087 TI - Infection control. Sharp shocks. PMID- 8424086 TI - Infection control. Dressing for protection. PMID- 8424088 TI - London sees rise in HIV among pregnant women. PMID- 8424089 TI - Critical aspects of initiation, promotion, and progression in multistage epidermal carcinogenesis. AB - Carcinogenesis in mouse skin can be divided into three distinct stages: initiation, promotion, and progression (malignant conversion). Initiation, induced by a single exposure to a genotoxic carcinogen, can result from a mutation in a single critical gene (e.g., rasHa), apparently in only a few epidermal cells. The change is irreversible. Promotion, resulting in the development of numerous benign tumors (papillomas), is accomplished by the repeated application of a nonmutagenic tumor promoter. The effects of single applications of tumor promoters are reversible since papillomas do not develop after insufficient exposure of initiated skin to promoters or when the interval between individual promoter applications is increased sufficiently. The reversibility of promotion suggests an epigenetic mechanism. Promoter treatment provides an environment that allows the selective clonal expansion of foci of initiated cells. The conversion of squamous papillomas to carcinomas (termed progression or malignant conversion) occurs spontaneously at a low frequency. The rate of progression to malignancy can be significantly increased by treatment of papilloma-bearing mice with certain genotoxic agents. These progressor agents or converting agents are likely to act via a second genetic change in papillomas already bearing the initiating mutation. Progression in the skin is characterized by genetic changes that result in several distinct changes in the levels or activity of structural proteins, growth factors, and proteases. The mechanisms involved in progression are being studied in epidermal cell culture. In order to determine the in vivo phenotype of cultured cells, a grafting system was developed in which the cells were transferred from culture to a prepared skin bed in athymic mice. Introduction of an activated v-fos oncogene into initiated cells bearing an activated rasHa gene produced cells with a carcinoma phenotype, i.e., carcinomas formed when the cells were grafted as part of reconstituted skin. Grafted keratinocytes containing the rasHa gene alone produced papillomas; with v fos alone, normal skin formed when grafted. The rasHa/fos carcinomas showed changes in differentiation markers characteristic of chemically induced carcinomas. A cell culture assay utilizing cells initiated by the introduction of an activated rasHa oncogene was developed to study progression. After exposure of initiated cells to progressor agents under conditions in which the proliferation of the rasHa-initiated cells was suppressed, proliferating foci developed, with a good correlation of activity in the assay with activity in the progression stage in vivo. The cell culture assay provides a quantitative model to study chemically induced neoplastic progression and may be useful to identify potential progressor agents. PMID- 8424090 TI - Manganese metabolism in rats: an improved methodology for assessing gut endogenous losses. AB - Manganese homeostasis is believed to be maintained by excretion of excess absorbed manganese through the gut, but the extent of endogenous gut losses of manganese has not been quantitated. We developed a model with rats to quantitate endogenous gut losses of manganese in which the parenterally administered isotope was distributed like fed isotope. Intraportally injected 54Mn complexed to albumin distributed in tissues like the fed isotope, but carrier-free 54Mn injected intraperitoneally, intravenously, or intraportally, or 54Mn complexed to transferrin and injected intraportally did not. Thus, manganese appears to be complexed to albumin or an albumin-like protein when it leaves the intestine. A mathematical model of manganese metabolism in rats fed 54Mn was developed using the SAAM and CONSAM computer programs. It was determined that the liver, not the pancreas, was the major source of endogenous gut losses of manganese. Young, growing rats fed 45 micrograms of Mn/g diet were calculated to absorb 8.2% of their manganese intake and then to lose 37% of the absorbed manganese through gut endogenous losses. PMID- 8424091 TI - Ontogeny of pituitary growth hormone and growth hormone mRNA in the chicken. AB - The changes in pituitary growth hormone (GH) mRNA levels have been determined by Northern blot analysis and laser densitometry during embryonic development and posthatch growth of white Leghorn cockerels. Pituitary GH mRNA levels were observed to progressively increase between 18 days of embryonic development to a maximum at 4 weeks of age (posthatch). Subsequently, pituitary GH mRNA levels declined between 4 and 8 weeks of age, and between 12 weeks of age and adulthood. Pituitary GH contents showed increases during embryonic development and posthatch growth that paralleled the rise in GH mRNA. The decline in pituitary GH mRNA levels between 4 weeks of age and adulthood occurs when GH secretion has been observed previously to decline. PMID- 8424092 TI - Influence of orotic acid on multistage hepatocarcinogenesis in the rat: resistance of hepatocytes from nodules to the mitoinhibitory effects of orotic acid. AB - This study was designed to determine whether hepatocytes from hepatic nodules are resistant to the mitoinhibitory effects of orotic acid. Hepatic nodules were initiated in Fischer 344 male rats with 1,2-dimethylhydrazine.2HCl (100 mg/kg ip) given 16 hr after two thirds partial hepatectomy and promoted by feeding a diet containing 1% orotic acid. Eight to 9 months later, when persistent nodules had developed, the rats were taken off the orotic acid diet and maintained on a semisynthetic basal diet for 2 to 5 weeks. The effect of orotic acid on the DNA synthesis in the hepatocytes isolated from hepatic nodules and from the surrounding nonnodular liver and from the age- and sex-matched control rats was studied. The results indicated that a dose of orotic acid (120 microM) that almost completely inhibited the transforming growth factor-alpha-induced DNA synthesis in hepatocytes from nonnodular surrounding liver and from age- and and sex-matched control liver could not inhibit the DNA synthesis in hepatocytes from hepatic nodules. These results are consistent with the postulate that orotic acid may promote liver carcinogenesis by a differential mitoinhibition of normal hepatocytes while permitting the initiated hepatocytes to respond to growth stimuli and form hepatic nodules. However, it needs to be determined whether differential mitoinhibition of normal hepatocytes is the mechanism by which orotic acid promotes liver carcinogenesis. PMID- 8424093 TI - Neoplastic progression of Syrian hamster embryo cells in culture. PMID- 8424094 TI - Stage of tumor progression, progressor agents, and human risk. PMID- 8424095 TI - Interferon-induced enhancement of transforming growth factor-alpha expression in a human breast cancer cell line. AB - Previous work from our laboratory has demonstrated that gamma-interferon (IFN) inhibits growth of the human breast carcinoma cell line MDA 468, while enhancing expression of epidermal growth factor receptor (EGFR). Epidermal growth factor at high levels is known to inhibit growth of this cell line. Because MDA 468 cells produce low levels of transforming growth factor (TGF)-alpha (a ligand for epidermal growth factor receptor), we reasoned that IFN-induced cytotoxicity could be partially mediated by enhanced secretion of TGF-alpha. Therefore, we determined the ability of IFN to modulate the endogenous expression of TGF-alpha by MDA 468 cells. IFN-gamma, at 500 units/ml, increased the levels of TGF-alpha in serum-free conditioned media of MDA 468 cells 3-fold as measured by radioimmunoassay. TGF-alpha mRNA was similarly increased approximately 3-fold after 5 days of IFN treatment as determined by dot blot and Northern analysis. IFN increased expression of TGF-alpha in conditioned media in a dose-dependent fashion. Increased secretion of TGF-alpha into conditioned media was not observed at Days 1 and 3. Similarly, increases in TGF-alpha mRNA were not observed at those time points. These results demonstrate that IFN-gamma enhanced secretion of TGF-alpha by MDA 468 cells. Although exogenous TGF-alpha inhibited MDA 468 cell growth, the role that the enhanced endogenous production of TGF-alpha plays in the cessation of cell growth induced by IFN remains to be determined. PMID- 8424096 TI - Constant light extends life in hamsters with heart disease. AB - Our previous work has shown that constant light can prolong the life of hamsters with heart disease. Although we have seen this result several times, constant light was not protective in our most recent experiment. We undertook this study because we had changed some conditions. As in previous experiments, we found that life in constant light extended life for cardiomyopathic hamsters as compared with others living in a 12:12-hr light:dark environment. A second factor affecting survival was the number of hamsters in a group. Hamsters housed in groups of five lived longer than those housed in groups of two under the same lighting conditions. Data from one experiment suggested that a short photoperiod (6:18 light:dark) was also protective. Although these experiments indicate that the protective effects of different light:dark schedules are not simple ones, they are important because their use may prove to be a helpful adjunct in the treatment of congestive heart failure. PMID- 8424098 TI - Renal nerves are involved in the natriuresis and diuresis produced by central administration of clonidine in the rat. AB - To determine whether renal nerves are involved in natriuresis or diuresis produced by the intracerebroventricular administration of clonidine (0.2, 2.0, and 8 micrograms/kg/min, and 2.0 microliters/min), urine flow, and sodium excretion were measured before and during clonidine administration from innervated and contralateral denervated kidneys in anesthetized (Inactin, 0.1 g/kg, ip) Sprague-Dawley rats. Baseline urine flow and sodium excretion were elevated after renal denervation prior to infusion of clonidine. Examining urine flow and sodium excretion before and during clonidine infusion indicated significant increases in urine flow and sodium excretion from the innervated kidneys but not from the denervated kidneys, possibly due to the renal sympatho inhibition in the innervated kidney. However, the higher doses of clonidine (2 and 8 micrograms/kg/min) may have diffused out of the intracerebroventricular space into the peripheral circulation and produced their effect by a direct action on the kidney. Subsequently, two experiments were performed to distinguish between a central action and peripheral action. First, clonidine was administered centrally with concurrent administration of an alpha 2-blocker, yohimbine (8 micrograms/kg/min, i.v.), peripherally. In a second experiment the dose of clonidine was reduced 10-fold such that this reduced dose did not produce a peripheral action but still produced the renal responses to central administration. The results of the latter two studies further confirmed that natriuresis and diuresis produced by intracerebroventricular administration of clonidine is in part mediated by renal nerves. PMID- 8424097 TI - Cytotoxicity of oxysterols on cultured smooth muscle cells from human umbilical arteries. AB - The lethal effect of 25- and 26-hydroxycholesterol on smooth muscle cells derived from human umbilical arteries was investigated. The extent of cellular death corresponded with increasing oxysterol concentrations and incubation times. Incubation of the cells with 0.5, 2.5, or 10 micrograms/ml of 25- or 26 hydroxycholesterol revealed a proportionality between the degree of cellular death and oxysterol concentration over the 5 days of the experiment. Correlation coefficients among the degree of cellular death, the exposing period, and oxysterol concentration were significantly different (P < 0.05). However, none of these changes were noted in the smooth muscle cells cultured for 5 days in a medium containing the same concentration of cholesterol or 0.5% ethanol. An increase in the concentration of either serum or cholesterol in the culture medium did not significantly reduce the cytotoxicity of 2.5 micrograms/ml of 26 hydroxycholesterol. The results of this study suggest that oxysterols have an injurious effect on arterial cells, and that the injurious effect could not be altered by cholesterol, which was present at a serum concentration 12 times higher than that of oxysterols. PMID- 8424100 TI - Effect of flow reduction on coronary blood flow heterogeneity. AB - The purpose of this study was to test the hypothesis that under reduced flow conditions, spatial heterogeneity (segment-to-segment variability) of myocardial blood flow would be inversely proportional to the rate of perfusion and that vasoconstriction would alter this relationship in anesthetized open-chest dogs. A carotid-left circumflex coronary artery shunt was created. Coronary blood flow determinations employed the radioactive microsphere technique. Fifty tissue samples were obtained from the affected left ventricle, including subepicardial and subendocardial samples. The coefficient of variation (CV = 100 x SD/mean), an index of spatial heterogeneity, was computed. Flows were obtained under three separate conditions: control, a 50% flow reduction in the circumflex artery, and vasopressin infusion, in which the administered dose reduced circumflex artery flow by 50%. The within-animal CV increased from a control of 18 +/- 9 to 45 +/- 25 with partial occlusion and 46 +/- 18 with vasopressin. No significant differences in CV were found between the two reduced flow conditions. A significant linear relationship between CV and mean coronary flow was found (r = 0.51, P < 0.001) over the entire range of flows studied (CV = -0.34*flow + 50.91) for control, 50% occlusion, and vasopressin treatment. The correlation was significantly improved (r = 0.69) using the equation CV = (1319.06/flow) + 4.44. Thus, the relative heterogeneity of coronary blood flow increased with reductions in coronary blood flow, regardless of the means of flow reduction. PMID- 8424099 TI - Effect of taurine levels on liver lipid metabolism: an in vivo study in the rat. AB - Previous studies using guinea pigs and cats have shown that liver lipid composition is affected by intrahepatic taurine levels. The purpose of the present study was to determine whether this sulfonated amino acid could also affect lipid metabolism in the rat, an animal capable of synthesizing substantial amounts of taurine and used extensively in studies on lipid metabolism. Wide variations in the hepatic taurine content were induced by administering either 1% taurine or 1% guanidinoethane sulfonate in the drinking water for 2 weeks. These treatments increased and decreased taurine liver content, respectively, but did not affect either food or water intakes, or growth rates. The plasma concentrations of the major lipid classes in treated animals did not show any significant alteration in comparison to control animals, except for nonesterified fatty acid levels that were significantly lowered by guanidinoethane sulfonate administration. Taurine supplementation did cause a significant decrease in total hepatic lipid content that was attributable to the reduction of free and esterified cholesterol, triglyceride, and phosphatidylethanolamine hepatic concentrations. This same treatment slightly increased both bile flow and secretion of taurine-conjugated primary bile salts. In particular, the proportion of tauro-beta-muricholate significantly increased, whereas that of taurodeoxycholate greatly decreased. The administration of guanidinoethane sulfonate reduced both the bile flow and the secretion of taurine-conjugated bile salts and caused a significant alteration in the ratio between glycine- and taurine-conjugated bile salts. This did not occur after the treatment with taurine. Interestingly, we observed an inverse correlation between hepatic taurine levels and the proportion of either cholesteryl ester in hepatic lipids or taurochenodeoxycholate in biliary bile salts. These facts suggest that taurine hepatic levels influence mostly hepatic steroid metabolism, but they also affect the metabolism of other lipid classes. PMID- 8424102 TI - Immunopathogenic mechanisms of posttransfusion graft-vs-host disease. AB - Posttransfusion graft-versus-host disease is a lethal adverse effect of blood transfusions affecting the skin, liver, gastrointestinal tract, and, in addition, the lymphohematopoietic systems. It is a disease that has been described for nearly three decades with well over 400 cases known worldwide; however, the immunopathogenesis has not been fully described. By using murine models, combined with human case reports, the immune mechanism and risk factors are outlined. The models and case reports prove a histocompatibility disparity between donor and recipient. The dose of lymphocytes in the products, types of T lymphocyte subsets in the products, and degree of immune suppression in the host are all factors necessary in the immunopathogenesis of posttransfusion graft-versus-host disease. The mechanism is that of acute, lethal, suppressive immune dysregulation in the host (recipient). PMID- 8424101 TI - Transcription factor GATA-1 and erythroid development. AB - In summary, we derived an experimental system that allows us to dissect the function of GATA-1 in red cell development at a genetic level. We have established the essential nature of GATA-1 during both primitive and definitive erythropoiesis. By ablating the expression of the endogenous GATA-1 gene, we are in a position to introduce a variety of constructs that harbor subtle modifications in flanking or protein-coding sequences. We can now study regulatory regions and functional domains of the protein in the context of a true erythroid environment, experiments that have not been possible heretofore. Although the assay involves the dramatic loss of red cell production, it should be possible to define important regulatory domains that can then be assayed using less stringent systems, such as cell-free extracts for in vitro transcription. The ideal situation would be analyses conducted in GATA-1- erythroid cells. However, these cells have been impossible to generate given the requirement of GATA-1 for Epo receptor expression and red cell viability (C. Simon and S. Orkin, unpublished observations). It may be possible to produce such cells by first expressing the Epo receptor under the influence of a constitutive promoter and then targeting the GATA-1 gene. If GATA-1- red cells were available, the analyses would involve the actual transcription of or chromatin structure surrounding the globin genes. Structure-function studies of the GATA-1 protein could be greatly simplified and a larger number of mutants studied. However, the ES cell system can be used as an alternative until targeted erythroleukemia cells become available. Other applications involve the introduction of other GATA-binding protein family members to determine whether they rescue the mutation. If they cannot, chimeric proteins can be tested to identify which amino acids distinguish the different family members. We feel that these experiments are vital to understanding the function of GATA-1 during erythroid ontogeny. How does GATA-1 regulate red cell genes like globin or the Epo receptor? Once we identify the functional domains of the GATA-binding proteins, we hope to learn what proteins GATA-1 binds to in the basic transcription machinery or in chromatin. Is GATA-1 necessary for globin gene switching? GATA-1 may be modified differently during development so that the locus control region can interact with different globin promoters. We may find that one region of the protein is required for embryonic expression and another for adult globin gene expression. PMID- 8424103 TI - Relaxin secretion by porcine large luteal cells: effect of protein synthesis inhibitors. AB - The purpose of the experiments reported herein was to investigate the relative importance of new hormone synthesis to basal and prostaglandin E2-stimulated rates of relaxin release. A relaxin-reverse hemolytic plaque assay was used to monitor relaxin release from individual large luteal cells (LLC) in which new protein synthesis was inhibited by cycloheximide or actinomycin D. These treatments significantly decreased the rate of relaxin release. In addition, cycloheximide reduced the total fraction of LLC possessing the ability to form plaques by about 10%, suggesting complete suppression of relaxin from this subset of cells. Exposure of inhibitor-treated LLC to prostaglandin E2 (a relaxin stimulatory secretagogue) enhanced relaxin release, and restored suppressed LLC back into the secretory population. Taken overall, these results demonstrate that the majority of relaxin-releasing LLC exploit a mixture of newly synthesized and older, stored hormone to achieve basal secretion. A minority of relaxin-releasing LLC, however, appear to depend wholly on newly synthesized hormone for basal secretion. The differential activity (and interaction) of these pathways in individual LLC may provide a potential explanation for the markedly heterogenous manner of hormone release observed in this (and other) cell types, and for the action of relaxin secretagogues. PMID- 8424104 TI - Lipopolysaccharide (LPS) processing by Kupffer cells releases a modified LPS with increased hepatocyte binding and decreased tumor necrosis factor-alpha stimulatory capacity. AB - Normal physiological clearance of gut-derived endotoxin lipopolysaccharide [LPS] has been described previously; initially, there is uptake by Kupffer cells (KC), then release of modified LPS, followed by hepatocyte uptake. Previous work in our laboratories indicated that LPS is structurally modified with loss of carbohydrate prior to its release by KC. In this study, we functionally characterize KC modified LPS. KC-modified 125I-LPS was prepared from primary rat KC. Escherichia coli 0127:B8 native 125I-LPS or KC-modified 125I-LPS (40 ng) was incubated for 1 hr with 1 x 10E6 primary hepatocytes. The binding of KC-modified LPS was 4.33-fold higher than native LPS (P = 0.0024). Binding analysis studies were conducted to determine the region of KC-modified LPS responsible for enhanced hepatocyte binding. KC-modified Salmonella minnesota LPS was competed with 100-fold excess native or mutant (Ra, Rc, Rd, or Re) strains of LPS or Lipid A with no decrease to hepatocyte binding. S. minnesota-native 125I-LPS was compared with KC-modified 125I-LPS in a study to assess induction of tumor necrosis factor (TNF)-gamma by rat peritoneal macrophages. Native or KC-modified 125I-LPS (100 ng) was presented to 1 x 10E7 peritoneal macrophages for 6 hr. TNF alpha was measured in supernatants using the WEHI-164 cytotoxicity assay. Native LPS induced 5.7-fold higher TNF-alpha levels than KC-modified LPS (P < 0.0001). The above data suggest that structural alterations in KC-modified LPS are accompanied by functional alterations resulting in enhanced hepatocyte binding and decreased TNF-alpha release. The latter result implies that an early step in LPS detoxification occurs in the KC in which LPS is modified to prevent elicitation of biologically active cytokines. PMID- 8424105 TI - Ligands influence Zn transport into cultured endothelial cells. AB - Experimental results were obtained that demonstrate the importance of the relative concentrations of Zn ligands and their affinities for Zn to the rate of Zn transport across a biological membrane. The transport rate into cultured endothelial cells became saturated near a Zn concentration of 30 microM in the presence or absence of 14% serum. However, the maximum transport rate identified by the saturation plateau was nearly twice as fast from serum-free medium. The addition of histidine or picolinic acid to the medium with serum resulted in the coincidental shift of Zn from high molecular weight ligands to low molecular weight ligands and increased the overall transport rate. In serum-free medium, adding histidine or picolinic acid slowed the transport rate. This indicates that the rate of Zn transport is influenced by the ligand to which it is associated and that altering the relative proportions of specific ligands influences the Zn transport rate. The rate of Zn transport decreased in a stepwise fashion as the albumin to Zn ratio increased from 0 to 4:1, with further increases having little effect. This suggests that albumin has a special role as modulator of Zn transport into endothelial cells. These studies underscore the importance of controlling the relative concentrations of Zn and its ligands in Zn transport kinetic research and suggest that varying their concentrations in a physiological range may be a method of regulating the distribution of Zn into specific cells and tissues. PMID- 8424106 TI - Thromboxane production in copper-deficient and marginal platelets: influence of superoxide dismutase and lipid hydroperoxides. AB - Platelet thromboxane (TX) production was examined in response to dietary copper. Groups of eight rats were fed copper-deficient, -marginal, and -adequate diets providing 0.5, 1.7, and 7.5 micrograms Cu/g, respectively, with controlled dietary Se and vitamin E. Platelets were purified and washed by centrifugation. Separate platelet samples from each rat were challenged with 10 micrograms/ml of collagen and 1 unit/ml (27.3 nM) of thrombin in Tyrode's buffer, 2.0 mM Ca2+. Platelet copper-dependent superoxide dismutase (CuSOD) activity showed a significant depression with reduced diet copper, but platelet glutathione peroxidase activity was unaffected. Challenged platelet TX production showed a significant 1.5- to 2.5-fold increase in response to both dietary copper deficiency and marginality, with highly significant negative correlations between challenged platelet TX production and platelet CuSOD activity and between TX production and copper status (liver copper). Endogenous (unchallenged) platelet lipid hydroperoxide concentrations, measured as free fatty acid hydroperoxides by a glutathione-disulfide-specific glutathione reductase recycling assay, showed a nonsignificant 47-67% increase in copper deficiency. Pooled data showed a significant 71% increase in platelet lipid hydroperoxides in copper deficiency. Platelet TX production showed a significant correlation with endogenous lipid hydroperoxides. The results suggest that dietary copper insufficiency increases platelet TX synthesis through changes in CuSOD in a dose-responsive (diet copper and platelet CuSOD activity) manner, and that platelet TX synthesis is influenced by lipid hydroperoxides (peroxide tone). PMID- 8424107 TI - Multiple pathways lead to activation of the survival mechanism in quiescent BALB/c-3T3 cells. AB - The survival of density-arrested quiescent murine BALB/c-3T3 cells in serum-free Dulbecco's medium requires the presence of cell growth factors or second messenger agonists. The protein synthesis inhibitor anisomycin blocks the survival-mediating action of the basic fibroblast growth factor (bFGF) and of 12 O-tetradecanoylphorbol 13-acetate (TPA), but has little or no effect on the protective action of platelet-derived growth factor or 8-bromoadenosine 3':5' cyclic monophosphate (Br-cAMP). The effects of anisomycin are concentration dependent in the range from 2.5 to 25 microM and show that the survival-enhancing abilities of bFGF and TPA critically require protein synthesis, whereas those of platelet-derived growth factor and Br-cAMP do not. The survival-mediating action of bFGF and TPA can also be blocked with the RNA synthesis inhibitors actinomycin D and 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), whereas the action of platelet-derived growth factor and Br-cAMP is largely resistant. Results on the time course of action of DRB, a selective inhibitor of the synthesis of mRNA precursor molecules, suggest that the RNA required for the survival-enhancing action of bFGF and TPA is present in cells at the time of serum withdrawal and addition of the survival factor and has a half-life greater than 3 h. The new evidence provides further support for the hypothesis that protection of serum deprived, density-arrested BALB/c-3T3 fibroblasts against death can be achieved either via pathways that entail the synthesis of protein and RNA (e.g., via diacylglycerol-protein kinase C) or via pathways that do not involve de novo biosynthesis (e.g., via cAMP-protein kinase A). PMID- 8424108 TI - Rotating-wall vessel coculture of small intestine as a prelude to tissue modeling: aspects of simulated microgravity. AB - A new low shear stress, low turbulence microcarrier culture system has been developed at NASA's Johnson Space Center that permits large-scale three dimensional tissue culture. Tissue culture bioreactors called rotating-wall vessels were used in conjunction with multicellular cocultivation to develop a unique in vitro tissue-modeling system. Normal small intestine epithelium and mesenchymal cells were cocultivated on Cytodex-3 microcarriers and were initiated in two phases. Normal small intestine mesenchymal cells were inoculated into the rotating-wall vessel at 2 x 10(5) cells/ml and allowed to attach and proliferate for 2 to 3 days. Normal small intestine epithelium was then added at an innoculum of 2 x 10(5) cells/ml and cultivation continued for 30 to 40 days. These cocultures attained cell numbers of 4-6 x 10(6) cells/ml and differentiated to form tissue-like masses of 0.4-0.5 cm with minimal necrosis. The masses displayed apical brush borders, differentiated epithelial cells, cellular polarity, extracellular matrix, and basal lamina. Verification of mesenchymal and epithelial cell expression was determined by immunocytochemistry and scanning electron microscopy. These data suggest that the rotating-wall vessel affords a new tissue culture model for investigation of growth, regulatory, and differentiation processes within normal tissues. PMID- 8424109 TI - Effects of estrogen and epidermal growth factor on prolactin and Pit-1 mRNA in GH3 cells. AB - The effects of epidermal growth factor (EGF) and 17 beta-estradiol (E2) on the expression of prolactin (PRL), the transcription factor Pit-1/GHF-1 (Pit-1), and on dopamine D2 receptor mRNA in GH3 cells were analyzed by immunocytochemistry, in situ hybridization, and Northern analysis in a defined serum-free cell culture medium. Radioimmunoassay was used to determine PRL secretion. Both EGF and E2 stimulated PRL mRNA and PRL secretion, although the effects of EGF were more rapid than those of E2. Pit-1 mRNA levels were not significantly changed in spite of the 2- to 8-fold increases in PRL mRNA levels and significant increases in PRL secretion. Analysis of dopamine D2 receptor mRNA by in situ hybridization and Northern hybridization detected expression of dopamine receptor in GH3 cells, but the receptor mRNA levels were not modified by EGF or E2 treatment in complete serum or in serum-free media. These observations suggest that EGF and E2 modulate PRL mRNA levels and PRL secretion significantly in vitro, while the mRNA levels of Pit-1 do not change significantly in GH3 cells cultured in a defined culture medium. PMID- 8424110 TI - The effect of neonatal sex hormone manipulation on the incidence of diabetes in nonobese diabetic mice. AB - The nonobese diabetic (NOD) mouse is a model of Type I (insulin-dependent) diabetes. It develops autoimmune pancreatic beta-cell lesions characterized by lymphocytic infiltration and beta-cell destruction. The incidences of diabetes for male and female NOD mice in our colony were 24% and 73%, respectively. In this study, we investigated the effect of neonatal manipulation of the sex hormone profile on the incidence of diabetes in male and female NOD mice. One day after birth, male mice were castrated and female mice were either ovariectomized, given testosterone, or ovariectomized and given testosterone. The mice were maintained for 140 days and blood samples were collected biweekly starting at 42 days old. Diabetes was determined by three consecutive blood glucose levels > 200 mg/dl. Neonatal gonadectomy increased the incidence of diabetes in males but decreased it in females. Females treated with testosterone also had a decreased incidence of diabetes, whereas ovariectomy plus testosterone increased the incidence to 100%. Castration decreased the body weight in males and increased body weight in females. Testosterone treatment with or without ovariectomy also increased body weight. From these studies, we concluded that neonatal hormonal imprinting has a significant influence on the incidence of diabetes in the NOD mouse. PMID- 8424111 TI - Polyamines and intestinal epithelial hyperplasia in streptozotocin-diabetic rats. AB - We measured specific activity of ornithine decarboxylase (ODC) and contents of putrescine and of the polyamines (spermidine and spermine) in isolated villus and crypt enterocytes from the jejunum of adolescent streptozotocin-diabetic and weight-matched control rats and diabetic and control rats treated with difluoromethyl ornithine (DFMO) 10 days after induction of diabetes. Consistent with previous observations by others of elevated ODC activity and contents of putrescine and of the polyamines in the intestinal epithelium undergoing hyperplasia, our studies showed elevated ODC activity and contents of putrescine and spermidine, but not of spermine, in the hyperplastic intestinal epithelium of diabetic rats. As in previous studies, suppression of ODC activity by DFMO prevented not only the jejunal epithelial hyperplasia in the diabetic rats, but also retarded jejunal epithelial growth in the control rats. DFMO administration lowered ODC activity by over 80% in both diabetic and control rat enterocytes and prevented the rise in enterocyte contents of putrescine and spermidine in the diabetic rat. The observation that, in both diabetic and control rats, treatment with DFMO lowered spermidine content in the crypt enterocytes but had no similar consistent effect on contents of putrescine or spermine suggested that spermidine could have been responsible for the intestinal epithelial hyperplasia in the diabetic rats and for the normal growth of the intestinal epithelium in control rats. PMID- 8424112 TI - Lacrimal gland uptake and metabolism of ascorbic acid. AB - Ascorbic acid is thought to contribute to protection against the potentially damaging effects of radiation, oxygen toxicity, and abrasion in the eye. The anterior surface of the cornea is particularly subject to insult from each threat. We considered the possibility that the lacrimal gland of pigs has transport and/or metabolic capability to sequester the reduced or oxidized form of ascorbic acid and prepare it for secretion in the tears. Slices of fresh lacrimal gland were incubated in a physiologic buffer and exposed to < or = 12 microM 14C-labeled ascorbic acid or dehydro-l-ascorbic acid over 40-min incubation periods. Dehydro-l-ascorbic acid was taken up to a greater extent than the reduced compound. 14C-Label recovered from the tissue was at least 75% in the form of ascorbic acid after incubation with either substrate. Uptake of both the reduced and oxidized substrates proceeded to a tissue to medium ratio in excess of unity; the former was prevented by the presence of nonlabeled ascorbate in the bathing medium, but was unaffected by the removal of Na+ from the bath. The uptake of both substrates was less after inhibition of cellular metabolic energy. The lacrimal gland in this diurnal animal species has transport and metabolic capabilities that could serve in secretion of ascorbic acid into tears. This might help to protect the corneal epithelium against various forms of damage. PMID- 8424113 TI - Contribution of plasma vasopressin concentration and blood pressure to norepinephrine-induced diuresis in conscious dogs. AB - Infusion of norepinephrine (NE) into humans and experimental animals induces diuresis by mechanisms that are not completely understood. Two series of experiments were performed to determine whether changes in plasma levels of vasopressin or changes in mean arterial pressure (MAP) are important-factors in NE-induced diuresis in conscious dogs. First, plasma vasopressin (PAVP) levels were measured in normal and cardiac-denervated conscious dogs during a 30-min intravenous infusion of NE (0.5 micrograms.kg-1.min-1). When NE was administered to normal dogs, urine flow increased from 0.3 +/- 0.1 to 0.9 +/- 0.4 ml/min. PAVP did not decrease, in spite of increases in mean arterial pressure (from 103 +/- 4 to 123 +/- 6 mm Hg) and left atrial pressure (from 5.2 +/- 0.9 to 8.6 +/- 1.4 mm Hg). The same dose of NE infused into cardiac-denervated dogs significantly increased urine flow (from 0.2 +/- 0.1 to 0.8 +/- 0.3 ml/min) and MAP (from 107 +/- 5 to 147 +/- 10 mm Hg), and decreased PAVP (from 1.8 +/- 0.3 pg/ml to 1.2 +/- 0.3 pg/ml). In the second series of experiments, NE was infused into cardiac denervated dogs for 40 min. During the final 20 min of NE infusion, nitroprusside was infused to offset the pressor effect of NE by returning MAP to the initial control level. Urine flow increased during the first 20 min in which NE alone was given; however, when MAP was returned to the control level by nitroprusside infusion, urine flow also returned to the control level. PAVP increased from a control value of 3.6 +/- 0.6 to 18.9 +/- 3.8 pg/ml 15 min after the NP infusion had begun. We conclude that a decrease in PAVP is not required to elicit diuresis during NE infusion in normal conscious dogs and that the pressor effect of NE appears to play a major role in NE-induced diuresis. PMID- 8424114 TI - Gastric toxicity and prostaglandin content in rats dosed with two chemically similar, nonsteroidal anti-inflammatory agents. AB - Two chemically similar nonsteroidal anti-inflammatory drugs, orpanoxin and F 1067, had almost identical potencies and efficacies as anti-inflammatory (rat paw edema) and analgesic (mouse writhing) agents, but differed markedly in gastrotoxicity. Orpanoxin alone aggravated stomach lesions in rats subjected to pylorus ligation and failed to protect stomachs of rats challenged with indomethacin. The compounds did not differ in their in vitro enzyme inhibition effects, both failing to inhibit 5- and 15-lipoxygenase and both inhibiting prostaglandin synthetase. Extraction of prostaglandins from the gastric mucosa of pylorus-ligated rats revealed, however, that the safer F-1067 depleted prostaglandin 6-keto-F1 alpha less and increased prostaglandin E2 much more than did orpanoxin. A possible causality is suggested. PMID- 8424115 TI - Monthly prostaglandin bibliography prepared by the University of Sheffield Biomedical Information Service. PMID- 8424116 TI - Preface--fatty acids and cell signalling. PMID- 8424117 TI - The role of free fatty acids in regulating the tissue availability and synthesis of sex steroids. AB - Sex steroids and dietary fat intake have been implicated in the growth of breast tumours. We have previously shown that the plasma free oestradiol fraction is increased in women with breast cancer and that the addition of free fatty acids (FFA) to plasma can increase the free oestradiol fraction in vitro. In the present study we have examined the distribution of oestradiol and testosterone in serum obtained from European women (EW) and Asian (Gujarati) women (GW) living in north-west London. Fat intake by these women is similar but GW, who are vegetarians, consume a greater proportion of unsaturated fats. In serum from perimenopausal GW, the free testosterone concentration was significantly higher than for EW (11.1 +/- 3.6 pmol/l vs 8.7 +/- 3.4 pmol/l, p < 0.05). Although a significant correlation was found between the free testosterone and FFA concentrations for GW (r = 0.49, p < 0.05), concentrations of sex-hormone binding globulin (SHBG) were significantly lower in GW than EW. The finding of lower SHBG concentration in GW was confirmed in a second study in postmenopausal women (EW, 60.1 +/- 34.1 nmol/l; GW, 37.8 +/- 20.5 nmol/l, p < 0.05). However, no difference in the free oestradiol fraction or concentration was detected for EW and GW and no correlations with total or individual FFA were found. It is concluded from this study that while dietary fats may have an important role in the development of breast tumours, it is unlikely to be mediated by FFA inhibiting the binding of sex steroids to plasma proteins. PMID- 8424118 TI - Physicochemical approach of the function of the fatty acid incorporation in biological membranes. PMID- 8424119 TI - Interactions between arachidonic and eicosapentaenoic acids during their dioxygenase-dependent peroxidation. AB - Eicosapentaenoic acid (EPA), a major polyunsaturated fatty acid of fish has been widely proposed as a potential nutrient for decreasing platelet-endothelial cell interactions and the subsequent atherogenesis and thrombogenesis. This is mainly based upon the decrease of arachidonic acid (AA) oxygenation into bioactive molecules like thromboxane A2. In addition, EPA may be oxygenated into its own active derivatives via cell dioxygenases. We report evidence for the requirement of specific peroxides, adequately provided by AA, to allow EPA to be oxygenated into its bioactive products like prostaglandin I3, a prostacyclin mimetic. On the other hand, we present some data that argue for a decreased basal AA dioxygenation (specific peroxidation) by small concentrations of EPA. The interactions between AA and EPA are then dual, EPA being able to counteract AA oxygenation whereas EPA requires AA to be efficiently oxygenated. PMID- 8424120 TI - Dynamics of the membrane lipid phase. AB - The lipid bilayer of a membrane is sometimes seen as an inert hydrophobic phase allowing the 'solubility' of transmembrane proteins and acting as a barrier between two compartments. However, the bilayer is, in fact, a highly organized system subjected to many movements leading to a dynamically equilibrated structure. A lipid within a membrane experiences intramolecular motions (movement of some segments of the molecule) and moves or diffuses in and across each monolayer. In plasma membranes, transverse diffusion is either passive (cholinecontaining phospholipids, fatty acids ...) or active via a carrier protein (amino-phospholipids). The known asymmetric transverse distribution of phospholipids between the two plasma membrane leaflets is a stationary state resulting from all these motions, especially the active transport. Nevertheless, recent studies have shown that it is also possible to obtain an uneven distribution of some lipids (e.g. fatty acid, phosphatidic acid) across a membrane via a pH gradient. Lateral diffusion within a monolayer depends on the composition of the monolayer and not on the nature of the diffusing lipid. The phospholipid asymmetry, based on the polar head groups, exists also for the corresponding fatty acids, as the nature of the acyl chains differs according to the head group. A consequence is that the cytoplasmic leaflet of plasma membranes has a different 'fluidity' from that of the outer leaflet. PMID- 8424121 TI - Cytoplasmic fatty acid binding protein: significance for intracellular transport of fatty acids and putative role on signal transduction pathways. AB - The cellular transport of long-chain fatty acid moieties is thought to be mediated by a plasmalemmal and a cytoplasmic fatty acid binding protein (FABPPM and FABPC, respectively) and a cytoplasmic acyl-coenzyme A binding protein (ACBP). Their putative main physiological significance is the assurance that long chain fatty acids and derivatives, either in transit through membranes or present in intracellular compartments, are largely complexed to proteins. FABPC distinguishes from the other proteins in that distinct types of FABPC exist and that these are found in a variety of tissues in remarkable abundance, with some cells containing more than one type In addition, liver type FABPC binds not only fatty acids, but also several other hydrophobic ligands, including heme, bilirubin, prostaglandin E1 and lipoxygenase metabolites of arachidonic acid. Calculations made for rat cardiomyocytes reveal that the presence of FABPC substantially enhances the cytoplasmic solubility as well as the maximal diffusional flux of fatty acids in these cells. Apart from this putative function in the bulk transport of ligands, FABPC may also function in the fine-tuning of cellular events by modulating the metabolism of hydrophobic compounds implicated in the regulation of cell growth and differentiation. PMID- 8424122 TI - Fatty acids may regulate aldosterone secretion and mediate some of insulin's effects on blood pressure. AB - Experiments in vitro and observation made in humans suggest that some unesterified fatty acids (FA) participate, as inhibitors, in the regulation of aldosterone secretion. Removal of FA from adrenal glomerulosa cells with albumin increases the responses to angiotensin II (AII) and dibutyryl cyclic AMP. Micromolar concentrations of some FA including arachidonic, oleic, linoleic, eicosapentaenoic, and docosahexaenoic inhibit aldosterone secretion by adrenal glomerulosa cells. Inhibition is specific--some acids like stearic are inactive, and the adrenal fasciculata is relatively resistant to inhibition. Oleic acid rapidly and reversibly inhibits aldosterone secretion by perfused dog adrenals. Observations in vivo suggest a reciprocal relationship between plasma levels of FA and aldosterone: insulin infusion into dogs lowers plasma FA and increases adrenal responsiveness to All; salt infusions into humans increase plasma FA as aldosterone falls; plasma FA are low in low-renin essential hypertension where adrenal responsiveness to All is high; plasma FA are inversely correlated with ratios of aldosterone to renin in black hypertensives; and plasma FA are high in some seriously ill patients whose aldosterone levels are inexplicably low. All receptors and the final step of aldosterone biosynthesis, oxidation at the 18 position, are the adrenal sites most sensitive to FA. Insulin's antinatriuresis may be mediated in part by its ability to lower plasma FA and thereby enhance adrenal response to secretagogues. PMID- 8424123 TI - Modulation of lipid derived mediators by polyunsaturated fatty acids. AB - Cell stimulation by a number of agonists triggers the formation of products of lipid hydrolysis, which act either as intracellular mediators of signal transduction or as modulators of cell-cell interactions. This process is mediated by the activation of hydrolytic enzymes, the phospholipases (PLase), especially the A2 and C, acting on cell phospholipids (PL). Among the major products being formed, the following: a) the inositol phosphates (IP), especially IP3, and diacylglycerols (DAG) generated intracellularly from phosphoinositides through PLase C, b) the eicosanoids, the arachidonic acid (AA) metabolites produced through combined PLase A2 and (cyclo- and lip-) oxygenase activities, and released from cells, c) the ether lipid PAF, derived from alkylacyl phosphatidylcholine through PLase A2, have attracted the attention of investigators for their important biological roles. Interest has also been recently developed towards products of sphingolipid hydrolysis, sphingosine and ceramide, which are generated by various cell types after stimulation, and exert biological activities. Cell glycerophospholipids are rich in long-chain polyunsaturated fatty acids (PUFA) of the n-6, namely AA 20:4 n-6, and n-3, mainly docosahexaenoic acid (DHA) 22:6, series. These compounds are differentially distributed among the various PL classes and their levels in cells are modulated through the intake with the diet of either the 18-C fatty acids (FA), precursors, linoleic 18:2 n-6, and, alpha-linolenic 18:3 n-3, respectively- followed by conversion to their long-chain PUFA derivatives, or through the intake of the performed compounds.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424124 TI - Fatty acids, inflammation and immune responses. AB - Evidence obtained from experiments in vitro and in vivo suggests that certain unsaturated fatty acids (FA) may be safe and effective antiinflammatory and immunomodulatory agents. Generation of a unique eicosanoid profile with different biological effects by administration of FA precursors other than arachidonic acid is one approach under investigation. In addition to their role as eicosanoid precursors, FA are of major importance in maintaining cell membrane structure, are key determinants of membrane bound enzyme activity and receptor expression. FA can exert these functions directly and therefore may themselves be important regulators of immune responses. For example, certain FA influence cytokine production and proliferation of human T lymphocytes in a manner that is direct and not due to their conversion to eicosanoids. The observations indicate that FA can modulate immune responses by acting directly on T-cells and suggest that alteration of cellular FA may be a worthwhile approach to control of inflammation. PMID- 8424125 TI - Role of polyunsaturated fatty acids as signal transducers: amplification of signals from growth factor receptors by fatty acids in mammary epithelial cells. AB - The growth, morphogenesis and differentiation of milk producing epithelial tissues in the developing mammary glands require interaction with extracellular matrices and stimulation by hormones, growth factors and essential fatty acids. In primary culture, the proliferation of mammary epithelial cells (MEC), induced by epidermal growth factor (EGF), is enhanced and sustained by linoleate and its eicosanoid metabolites. Since a combination of linoleic acid (18:2 omega 6) and prostaglandin E2 or cAMP has synergistic effect on EGF-stimulated growth, it is suggested that additional cAMP-dependent protein kinase A (PK-A) independent pathways may also contribute to the linoleate effect on EGF action. Possible involvement of Ca2+/phospholipid-dependent protein kinase C (PK-C) is explored. Both linoleate and arachidonate can activate Type-II and Type-III protein kinase C in MEC and a PK-C inhibitor can block growth stimulation by EGF and fatty acids. Like 12-O-Tetradecanoly phorbol-13-acetate (TPA), a PK-C activator which also enhances EGF-stimulated growth of MEC, linoleate can phosphorylate a 40-42 KD protein. EGF itself can stimulate transient phosphorylation of the same protein in MEC cultures but when supplemented with linoleate, which does not influence the ligand binding affinity of EGF-receptors, the transient phosphorylation signal in 40-42 KD protein is sustained.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424126 TI - Arachidonic acid and cell signalling in the ovary and placenta. AB - Arachidonic acid (AA) and its metabolites make up a diverse group of signalling molecules important to mediation of metabolic and endocrine function of ovarian and placental cell membranes. This paper reviews recent literature examining AA and eicosanoid involvement in the functional dynamics of follicular development, ovulation and corpus luteum function. The putative roles of AA metabolites in establishment and maintenance of pregnancy are reviewed with reference to decidualization, trophoblast invasion and implantation, maintenance of perfusion of the feto-placental unit and lipid transfer. Finally, recent evidence implicating AA metabolism in mediation of enzyme activity following hormone receptor coupling within various cells types comprising the placental membranes is reviewed. PMID- 8424127 TI - Regulation of gene expression by fatty acids in the adipose cell. PMID- 8424128 TI - The cerebellar olivo-corticonuclear connections in the rat. PMID- 8424129 TI - Indomethacin inhibits cell proliferation in the oxyntic epithelium of the rat. AB - The aim of this investigation was to examine the action of parenteral indomethacin and oral prostaglandin E2 on cell proliferation in the rat oxyntic mucosa. Groups of Sprague Dawley rats were treated with either 1.5 mg/kg indomethacin subcutaneously, 5 mg/kg oral prostaglandin E2 or placebo, twice daily during 5 days. All rats were killed exactly 4 hours after mitotic arrest with vincristine, and a biopsy specimen from the oxyntic mucosa was processed for routine microscopic evaluation. Mitotic figures were distributed cluster-like along the oxyntic mucosa alternating with mitosis-free areas. The total number of mitotic figures in 8 mm of mucosa was significantly reduced by administration of indomethacin (p < 0.05). In rats given indomethacin, 32.5% of the examined mucosa did not have mitotic figures, which is significantly higher than 14.3% as observed in placebo-treated rats (p < 0.05). Both rats treated with indomethacin and with prostaglandin E2 had fewer microscopic fields containing 5-6 mitotic figures than placebo-treated animals (p < 0.05). The maximal length of mitosis free areas was 0.6 (0.6-0.9) mm in rats given indomethacin which is significantly larger than 0.4 (0.2-0.4) mm observed in controls (p < 0.05). Indomethacin produced epithelial atrophy as shown by a significant reduction of the epithelial height observed in those rats compared to controls (p < 0.05). The inhibition of cell proliferation observed in the oxyntic mucosa of rats treated with the cyclooxygenase blocker indicates that an important physiological role of endogenous prostaglandin is to maintain the proliferative activity of the epithelium at a high level. PMID- 8424130 TI - Transforming growth factor-beta potentiates epidermal growth factor-induced prostaglandin E2 production in amnion cells. AB - Primary cultures of human amnion cells and the amnion-derived cell line WISH were used to evaluate the hypothesis that transforming growth factor-beta (TGF-beta) can modulate epidermal growth factor (EGF)- induced prostaglandin E2 (PGE2) production. Cells were preincubated for 1 hr with TGF-beta (0.0001-10 ng/ml) and then incubated in the presence or absence of EGF (10 ng/ml) for 4 hrs. TGF-beta alone did not stimulate PGE2 synthesis at any dose examined. However, when primary cultures of amnion cells or WISH cells were preincubated with TGF-beta and then challenged with EGF, there was a potentiation of PGE2 production that was much greater than the additive values of TGF-beta or EGF alone. These data suggest that EGF-induced PGE2 production by amnion cells can be modulated by low concentrations of TGF-beta. PMID- 8424131 TI - Thromboxane synthetase inhibition as a new therapy for preeclampsia: animal and human studies minireview. AB - The role of the eicosanoids in the pathophysiology of preeclampsia is reviewed, and the results of animal model and human studies with thromboxane synthetase inhibitors in preeclampsia are described. Potential benefits and limitations of therapy are discussed. PMID- 8424132 TI - Maternal and fetal prostaglandin concentrations during late gestation in dairy cattle. AB - A study was conducted to measure the blood plasma concentrations of prostaglandin F2 alpha (PGF2 alpha), 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM), prostaglandin E2 (PGE2) and 13,14-dihydro-15-keto-prostaglandin E2 (PGEM) in the jugular vein, umbilical vein and artery and uterine vein of 18 Holstein Friesian cows during late gestation. A caesarean section was performed on all cows before term in order to obtain blood samples from the different sources. Plasma PG concentrations in the uterine or fetal circulation were significantly higher than in jugular vein plasma. Correlations between peripheral PG metabolite concentrations and primary PG concentrations in the various sources of the uterus or fetus were not significant (r = .17-.47) and demonstrated that prostaglandin values based upon peripheral blood alone are of limited value. PMID- 8424133 TI - The effect of thromboxane A2 receptor antagonism on amphotericin B-induced renal vasoconstriction in the rat. AB - Amphotericin B (AmB) is the drug of choice for most systemic fungal infections, but doses are frequently reduced because of nephrotoxicity. We investigated the role of thromboxane as a mediator for this nephrotoxicity. Vehicle or amphotericin (0.60 mg/kg) was infused into the left renal artery in four groups of rats, and renal plasma flow (RPF) and glomerular filtration rate (GFR) were measured. Group 1 received vehicle for 90 min. Group 2 received vehicle followed by a 30 minute AmB infusion which caused a significant and reversible fall in the RPF and GFR. Group 3 received vehicle followed by AmB infusion, but were infused with a bolus of ibuprofen (20 mg/kg) 45 minutes before AmB. This group exhibited an insignificant attenuation in the fall in RPF and GFR. Group 4 received vehicle followed by AmB, but were infused with a bolus and continuous infusion of the thromboxane receptor antagonist SQ29,548. This group demonstrated an attenuation in the fall in RPF and a significant decrease in GFR compared to AmB control rats. In addition, the rat glomeruli were incubated with AmB (4 ug/ml). Supernatant levels of thromboxane B2 were significantly elevated in the presence of AmB vs buffer alone. We conclude that the reduction in RPF and GFR observed with AmB infusion in the rat is partially mediated by release of thromboxane. PMID- 8424134 TI - Antagonist of platelet-activating factor prevents prostaglandin E2 induced ocular hypertension in rabbits. AB - Topical application of 250 micrograms of platelet activating factor (PAF) or 5 micrograms of prostaglandin E2 (PGE2) to the rabbit's eye is followed by a rise in intraocular pressure (IOP). The occurrence of PAF in the anterior chamber is held responsible for this effect, as appears from the observed concentrations of PAF in a ng/ml-range in the aqueous humor and from the inhibition in the rise of IOP by systemic pretreatment of rabbits with the PAF-antagonist BN 52021 (10 mg/kg). Corneal and conjunctival edema, occurring after application of PAF, are not influenced by BN 52021. Thus, these effects may be ascribed to the non specific membrane pertubant action of the high local concentration of PAF, necessary to achieve effective concentrations of PAF in the anterior chamber. PMID- 8424135 TI - [Anxiety, a basic human experience]. AB - Anxiety, unlike fear, which is strongly object-related, accompanies man all his life. This anxious condition appears especially then, when danger is imminent, also in the case of being severely ill. Anxiety and hope, even in people severely ill, do not exclude each other. Human life for many and perhaps for all is a race with time. The anxiety not being able to realize enough in a given time often is the reason for a busy and restless activity. Man often suffers from anxiety of other human beings. As ever an individual may live, it always owes something to others. Accordingly, the doctor or other helpers never are able to behave totally in the way as the patient expects and the patient cannot act as the helper truly would like it. Man experiences anxiety, however, also then, when he recognizes to remain in debts towards himself and, in the view of religion, towards a task given by God. Yet, man suffers anxiety not only because of his confrontation with death but also with life and the responsibilities which he has to take over when growing up in society. The struggle for existence described by Darwin, a result of anxiety, seems to be lost for the patient knowing he or she is severely ill, and he/she is therefore upset against a destiny which threatens to exclude him/her from the community of healthy people and perhaps soon from the living persons. Anxiety always turns also around the existence in the group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424137 TI - [Opinion on the Swiss AIDS Clarification on informed consent and HIV testing]. PMID- 8424136 TI - [Anxiety and terminal tumor disease]. AB - Clinically relevant anxiety disorders are rare in terminally ill cancer patients. A variety of factors can cause symptoms of anxiety in this patient population, the imminence of death is not the most frequent reason. According to the results of a psychiatric evaluation, the physical examination and further diagnostic procedures, treatments can be effective in alleviating anxiety in these patients. PMID- 8424138 TI - [Patient without being ill? Benefit and risks of presymptomatic diagnosis]. AB - A steadily growing number of genetic disorders or for predispositions disease can already be identified prior to their clinical manifestation. Gene technology has a considerable impetus on the development of presymptomatic or preclinical testing. Using familial cancer as a model it will be shown that persons at risk can be protected from the severe consequences of their genetic traits and can be counselled with certainty with respect to their family planning. However, such hopes are accompanied by fears that this type of genetic analysis will be performed without appropriate genetic counselling, without accepted medical practices or without the necessary quality control and that the results may be misinterpreted or misused by third parties. Informing the medical profession and the public about presymptomatic testing is the best way to prevent such negative consequences. PMID- 8424139 TI - [Autonomy of the patient and the physician in resuscitation situations and in terminal care. The value of advance directives by the patient]. AB - Interactions between the autonomy of the patient and that of the physician (or his professional ethics respectively) in difficult situations, i.e. resuscitation or care in terminal phases of disease, are discussed. In this respect the ethical and legal value of a declaration written and signed by the patient at a time of good health (will) is analyzed. PMID- 8424140 TI - [A case from practice (260). Moderate to highly differentiated follicular thyroid carcinoma]. PMID- 8424141 TI - [If one heeds the general practitioners?]. PMID- 8424142 TI - [A view of general practice]. AB - What happens during our consultation and what are the determining factors or specific practice patterns in general practice? Six Swiss general practitioners discuss this questions based on a random sample of 300 descriptions of unbiased consultations. Further information obtained from the patient after the consultation by questionnaire and a recorded semi-structured interview was analysed. In the following article the methods are presented. PMID- 8424143 TI - [Is there a typical general practitioner?]. AB - A random sample of 300 unbiased consultations on the premises of six Swiss general practitioners was analysed. In this article the results of the 'hard facts' (from patient questionnaires and the physicians tape-recorded consultation's essentials) are referred. The consultation length (average 16.5 min.) was underestimated by doctor and patient in 64% and 41% respectively. 81% of the patients saw us as their 'family doctor'. This term, however, is badly defined. Previous visits at home had taken place in 26 to 54% of all cases. Full patient satisfaction with the consultation was found in 80 to 90%, and the patients attested an optimal emotional approach. Also the financial aspects are mentioned. The authors are impressed by the finding of significant inter-doctor differences of important physician's practice patterns. PMID- 8424144 TI - [Significance of diagnostic aspects in general practice]. AB - The role of the 'diagnosis' in general practice is different from that in teaching hospitals, but a broader definition is needed. In general practice the spectrum of diagnoses appears to be almost unlimited ('for every patient his own diagnosis'). Therefore, the author calls in question any classification system. In a personal study of 300 consultations in the offices of six Swiss general practitioners, no main diagnosis was reported in 16%, and in one third the main diagnosis was uncertain. The impact of the diagnosis on the therapeutic consequences or on prognosis seems to be overestimated. Of the stated diagnoses most were somatic, but not in the sense of 'clinical' entities. Only few patients are interested in our diagnostic terms. Significant inter-practitioner variations in the diagnostic approach are discussed. PMID- 8424146 TI - [Clarity in general practice]. AB - A random sample of 224 consultations and interviews was analyzed, regarding the structuring and sequences in each consultation. Most space in the unbiased doctor's descriptions took anamnesis, followed by therapy, investigation, followed by physical examination and, least, diagnosis. Urban doctors kept the classic scheme of succeeding anamnesis, investigation, diagnosis and therapy more often than their rural colleagues. Explanations to patients were more frequently done by country doctors. The words doctors used to indicate communication are listed. The term of resonance phase in consultation is discussed. To achieve clarity in the consultation is important for doctor's stress prophylaxis. PMID- 8424145 TI - [Feelings in general practice]. AB - 300 random-sampled consultations were described by general practitioners and by interviews of the patients. Emotional aspects from both parts were analysed and correlated with patient centredness. We found significant variation, related to practitioner patterns, whereas patient's satisfaction alone showed no significant difference. PMID- 8424148 TI - The state of South Dakota's child: 1992. AB - This annual report on South Dakota's children shows that the state's birth rate in 1991 has again dropped and is below the national rate. In recent years there has been progress noted in the utilization of first trimester prenatal care and the state's infant mortality rate is comparable to that observed nationwide. Injuries, the leading cause of death in childhood, is the focus of this year's report. The rate of death due to childhood injuries is higher in South Dakota than is observed nationwide. Injuries are responsible for 51% of all deaths among 1 to 4 year olds, 70% of those of 5 to 14 year olds and 82% of all deaths of those 15 to 19 years of age. Strategies for the prevention of this tragic loss of life are discussed and the continued need for broad-based educational efforts and public policy that promote safe habits is described. PMID- 8424149 TI - Systemic effects of ophthalmic agents for the treatment of glaucoma. PMID- 8424150 TI - Bartholin cyst presenting as inguinal hernia. AB - The clinical diagnosis of Bartholin cysts is usually very simple. This paper, however, reports a very unusual presentation, a Bartholin cyst originally thought to be an inguinal hernia. The history, anatomy, and clinical findings of Bartholin cysts are discussed, as well as a case history of this interesting presentation. PMID- 8424147 TI - [Do we use the potential of the general medicine consultation?]. AB - Is there any congruence between vocational training as general practitioner and the needs of patients in primary care in Switzerland? What are the respective needs in the next century? These questions are discussed, based on the personal experience of the author after having completed his vocational training as general practitioner, on three years of experience as research fellow in a descriptive, cross-sectional and observational study of 300 consultations on the premises of six general practitioner-investigators and on a review of key research literature. Controversial views on the potential and the gaps in today's primary care are shown, focused on definition, training and research in the field of general or family practice. Of a broad range of experiences, two are pointed out: significant interindividual differences in practice patterns in primary care and emerging differences between primary care and teaching hospital medicine. The exceptional potential of primary care consultations has much to contribute to needed reforms in medical education and the health care system for the 21st century. Primary-care-based research and training will develop improved skills e.g. in communication, medical decision-making and patient management. Primary care is a discipline which includes insecurity and individuality; therefore, open questions and constant changes will be part of present and future for us and our patients. PMID- 8424151 TI - FNA--how can it best be used? PMID- 8424152 TI - Cadmium and lead in the smoke of a filter cigarette. AB - Cadmium and lead were determined in the parts of a filter cigarette after fractional smoking. The partitioning of these elements into the main smoke stream, consisting of particulates and gases, also into the side stream, the ash and the butt was determined. Approximately 70% of the lead mobilized by the smoking process was found in the ash; about 50% of the cadmium was transferred into the side stream. The overall trend shows an increase in the amount of the metals in the main stream as the number of puffs is increased. A model for the behavior of the main stream components within the cigarette was developed and showed a high analogy with the Lambert-Beer law describing the absorption of light. PMID- 8424153 TI - Microbiological quality of recreational waters in Araraquara, SP, Brazil. AB - The microbiological flora of 108 water samples was explored to evaluate the role of recreational waters as a possible source of human diseases in Araraquara, S.P., Brazil. These waters included six swimming pools and three lakes with beaches. The number of total and fecal coliforms, Escherichia coli, fecal streptococci, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and heterotrophic organisms was determined. As was the occurrence of Salmonella, Shigella, yersinia, enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Mycobacteria, yeasts in general and dermatophytes. Shigella, Yersinia, EIEC and dermatophytes were not isolated. Other organisms or groups of microorganisms were found in variable proportions. From this study it is concluded that recreational waters used by the population of Araraquara, may be contaminated with potentially pathogenic microorganism and this may serve as a source of human diseases. PMID- 8424154 TI - Effect of leachate from landfills on underground water quality. AB - A study was performed in order to evaluate the effect of leachate from a landfill, in the Athens region, to the underground water of the surrounding area. A number of physical chemical parameters were monitored, including metals from various sampling sites, covering an area up to 3 km from the landfill. The experimental results indicate that the underground water quality has been greatly affected by the leachate, since most of the parameters examined show increased concentrations. PMID- 8424155 TI - Zinc, copper, and selenium in milk and organs of cow and goat from Burundi, Africa. AB - Atomic absorption spectrometric determination of zinc, copper and selenium in milk from cow and goat from Burundi provide values, which agree quite well with literature data. Muscle from both animals is a rich source of zinc, but not for copper and selenium. Liver accumulates the highest amount of copper, while for selenium the kidney is the target organ. PMID- 8424156 TI - Impact of repackaging hazardous (infectious) hospital waste on the indoor air quality of a hospital. AB - With landfill bans on the disposal of hazardous (infectious) hospital wastes, many hospitals in the United States are faced with selecting alternative means of disposal. One alternative method of disposal is to repackage the waste and ship it off site for incineration. This paper reports on a study in which airborne bacteria were sampled in an area where waste was repackaged. Results of the sampling showed bacteria concentrations ranging from not detectable (central supply room) to 105 colonies/m3 air (approximately 40 feet from the incinerator room door). This would seem to indicate that the source of the bacteria was at or near the incinerator room door where the repackaging operation took place. Although some human pathogens were identified, most of the bacteria identified do not represent a serious health threat. However, Staphylococcus aureus was found in air samples collected from locations in the hospital served by the same ventilation system in the area of the repackaging operation. The waste handling practices used in hospitals need to be reevaluated, and the definition of 'sick building syndrome' needs to be expanded beyond chemicals to include microorganisms. More research is needed to characterize bacteria in air and to determine the impact of airborne bacteria on human health. PMID- 8424157 TI - Relationship between the elemental content and pH of human hair. AB - In this study, significant correlations were discovered between the following three parameters: (i) pH values of human hair, (ii) K, Ca, Mg, Zn contents of human hair and (iii) sex. We also discovered that most trace elements are not significantly correlated with hair pH. PMID- 8424158 TI - Indoor radon-daughter concentration and gamma radiation in urban and rural homes on geologically varying ground. AB - The radon (Rn)-daughter level (track film, 3 months) was lower in 129 urban than in 197 rural houses (geometric means, (GM) 18 versus 40 Bq/m3; P < 0.001; range 3 416). In rural dwellings higher levels were found in those built over deposits of alum shale occurrences (P = 0.04; 12 on porous glaciofluvial cover 94 Bq/m3; 127 others 43 Bq/m3; significant interaction, P = 0.01), than in 158 others (37 Bq/m3). There was no systematic effect of the type of water supply, neither of indoor smoking. One hundred twenty three homes with a complete cellar were lower than 200 with no or part cellar (18 versus 40 Bq/m3; P < 0.001). There was only a weak association between log-transformed gamma radiation (GM 10, range 33-50 microR/h) and Rn-daughter levels (r = 0.18; 95% confidence interval 0.07-0.28). Our results show the importance for indoor Rn-daughter levels of the bedrock and cover beneath the house, and the need for Rn-daughter measurements in epidemiological studies. PMID- 8424160 TI - Measles vaccine: titre and safety. PMID- 8424159 TI - Business and science. PMID- 8424161 TI - Fetal transplant update. PMID- 8424162 TI - Science and the new administration. PMID- 8424163 TI - A grisly archive of key cancer data. PMID- 8424164 TI - Radiation risks. Researchers eager to see soviet data. PMID- 8424165 TI - Should dying patients receive untested genetic methods? PMID- 8424166 TI - Scientific publishing. Will the real journal please stand up? PMID- 8424167 TI - Alzheimer's pathology begins to yield its secrets. PMID- 8424168 TI - Catalytic antibodies and disfavored reactions. PMID- 8424169 TI - The polar coordinate model goes molecular. PMID- 8424170 TI - A wingless-dependent polar coordinate system in Drosophila imaginal discs. AB - The patterning of the imaginal discs in Drosophila melanogaster is a progressive process that, like the patterning of the larval epidermis during embryogenesis, requires the activity of segment polarity genes. One segment polarity gene, wingless, encodes a homolog of the mouse proto-oncogene Wnt-1 and plays a prominent role in the patterning of the larval epidermis and the imaginal discs. However, whereas the function of wingless in the embryo is initially associated with a pattern of stripes along the anteroposterior axis that are part of a Cartesian coordinate system, it is shown here that during imaginal development wingless is associated with a pattern of sectors that provide references for a polar coordinate system homologous to that postulated in a well-known model for the regeneration of insect and vertebrate limbs. PMID- 8424171 TI - Antibody catalysis of a disfavored chemical transformation. AB - Organic reactions are often limited by stereoelectronic constrains that appear along the reaction coordinate. An antibody has been generated that overcomes these constraints and catalyzes a highly disfavored chemical transformation. The antibody facilitates the difficult 6-endo-tet ring closure of an epoxy-alcohol to form a tetrahydropyran. The catalyzed process is in formal violation of what has become known as Baldwin's rules for ring-closure reactions. In addition to controlling the regiochemistry of the disfavored cyclization reaction, these catalytic antibodies resolve enantiomeric substrates to afford a stereochemically pure product. The principles demonstrated in this study may be applicable to other disfavored chemical processes. PMID- 8424172 TI - In situ stimulation of aerobic PCB biodegradation in Hudson River sediments. AB - A 73-day field study of in situ aerobic biodegradation of polychlorinated biphenyls (PCBs) in the Hudson River shows that indigenous aerobic microorganisms can degrade the lightly chlorinated PCBs present in these sediments. Addition of inorganic nutrients, biphenyl, and oxygen enhanced PCB biodegradation, as indicated both by a 37 to 55 percent loss of PCBs and by the production of chlorobenzoates, intermediates in the PCB biodegradation pathway. Repeated inoculation with a purified PCB-degrading bacterium failed to improve biodegradative activity. Biodegradation was also observed under mixed but unamended conditions, which suggests that this process may occur commonly in river sediments, with implications for PCB fate models and risk assessments. PMID- 8424173 TI - Altered specificity of DNA-binding proteins with transition metal dimerization domains. AB - The bZIP motif is characterized by a leucine zipper domain that mediates dimerization and a basic domain that contacts DNA. A series of transition metal dimerization domains were used to alter systematically the relative orientation of basic domain peptides. Both the affinity and the specificity of the peptide DNA interaction depend on domain orientation. These results indicate that the precise configuration linking the domains is important; dimerization is not always sufficient for DNA binding. This approach to studying the effect of orientation on protein function complements mutagenesis and could be used in many systems. PMID- 8424174 TI - Release of excess amyloid beta protein from a mutant amyloid beta protein precursor. AB - The 4-kilodalton amyloid beta protein (A beta), which forms fibrillar deposits in Alzheimer's disease (AD), is derived from a large protein referred to as the amyloid beta protein precursor (beta APP). Human neuroblastoma (M17) cells transfected with constructs expressing wild-type beta APP or a mutant, beta APP delta NL, recently linked to familial AD were compared. After continuous metabolic labeling for 8 hours, cells expressing beta APP delta NL had five times more of an A beta-bearing, carboxyl terminal, beta APP derivative than cells expressing wild-type beta APP and they released six times more A beta into the medium. Thus this mutant beta APP may cause AD because its processing is altered in a way that releases increased amounts of A beta. PMID- 8424175 TI - Activation of the sphingomyelin signaling pathway in intact EL4 cells and in a cell-free system by IL-1 beta. AB - The mechanism of interleukin-1 (IL-1) signaling is unknown. Tumor necrosis factor alpha uses a signal transduction pathway that involves sphingomyelin hydrolysis to ceramide and stimulation of a ceramide-activated protein kinase. In intact EL4 thymoma cells, IL-1 beta similarly stimulated a rapid decrease of sphingomyelin and an elevation of ceramide, and enhanced ceramide-activated protein kinase activity. This cascade was also activated by IL-1 beta in a cell-free system, demonstrating tight coupling to the receptor. Exogenous sphingomyelinase, but not phospholipases A2, C, or D, in combination with phorbol ester replaced IL-1 beta to stimulate IL-2 secretion. Thus, IL-1 beta signals through the sphingomyelin pathway. PMID- 8424177 TI - On the relationship between knowledge and action in health policy. PMID- 8424178 TI - Women, men and health. Introduction. PMID- 8424179 TI - Special issue. Women, men and health. PMID- 8424176 TI - Regulation by heme of mitochondrial protein transport through a conserved amino acid motif. AB - A conserved motif, termed the heme regulatory motif (HRM), was identified in the presequences of the erythroid delta-aminolevulinate synthase precursors and was shown to be involved in hemin inhibition of transport of these proteins into mouse mitochondria in vitro. When the HRM was inserted into the presequence of the ornithine transcarbamoylase precursor, a normally unregulated mitochondrial protein, it conferred hemin inhibition on the transport of the chimeric protein. The conserved cysteine within the HRM was shown by site-directed mutagenesis to be required for hemin inhibition. PMID- 8424180 TI - Gender differences in the perceptions of common cold symptoms. AB - Higher rates of reported morbidity among women are sometimes attributed to lower thresholds among women for experiencing and reporting symptoms. Gender differences in the perception of signs and symptoms of minor illness were examined on data from the MRC Common Cold Unit. Volunteers assessed the presence and severity of colds at the end of their stay in the Unit, using the same two measures as a trained clinical observer (all ratings were double blind). Even after adjustment for other variables, men were significantly more likely to 'over rate' their symptoms in comparison with the clinical observer than were women. These data, and recent analyses from elsewhere, suggest that rather than artefactually exaggerating gender differences in morbidity, differing thresholds for perceiving and reporting symptoms may produce underestimates of gender differences in morbidity. PMID- 8424181 TI - Gender inequalities in health: social position, affective disorders and minor physical morbidity. AB - Research has consistently reported a female excess of minor physical morbidity and affective disorders compared to men. Using data from a national cross sectional survey of British women and men aged 18-59, this paper explores three prominent explanations for these patterns: that the female excess is due to problems associated with menstruation and the menopause; that it is 'caused' by the social positions which women occupy in contemporary western societies; and that the excess of affective disorders is 'caused' by women's social positions but that their higher rate of physical illness is psychosomatic in origin. The analysis presented here suggests that problems with menstruation and the menopause cannot explain gender inequalities in minor illness. Similarly the argument that the female excess of minor physical illness is psychosomatic is not supported, indeed, there is evidence that women's experience of minor physical illness may 'cause' their higher rates of affective disorders. Finally, with a few exceptions, the relationship between minor illness and four dimensions of social positions--marital status, employment status, social class and income- were broadly the same for women and men but in each social position category, women report higher rates of both types of ill health than men. It is concluded that present measures of these social positions do not capture the differing experiences of women and men and that more gender sensitive measures are needed if gender inequalities in minor illness are to be understood. PMID- 8424182 TI - Gender and inequalities in health in later life. AB - Little research attention has been given to examining inequalities in the health of elderly women and men, in spite of their high use of health services and the importance of health to maintaining independence in later life. This paper uses data from the British General Household Survey to analyse the variation in health of elderly women and men by class and material circumstances. Two measures of health are used; self-assessed health and functional disability. Elderly women assess their own health less positively than men, and are seriously disadvantaged compared to men in terms of functional disability. Class based on the individual's own previous main occupation is strongly associated with the two measures of health for elderly women and men at all ages. For elderly women, an 'individualistic' approach, using the woman's own last occupation, is compared with the 'conventional' approach of measuring class, which for married women uses their husband's last occupation and for other women their own last occupation. Using the two approaches makes little difference to the strength of association between class and health. Elderly women and men who live in advantaged material circumstances, in terms of income, car ownership and housing tenure, report significantly better health, after controlling for age and class. Level of functional disability is influenced by previous position in the labour market but not current material circumstances. Although elderly women suffer greater morbidity than elderly men, structural inequalities in health are equally pronounced for women and men in later life. PMID- 8424183 TI - Prostitutes and chimney sweeps both have problems: towards full integration of both sexes in the study of occupational health. AB - Recently, natural and social scientists have pointed out that the need to make scientific results apply to both sexes is not met by simply adding women as research subjects. They suggest that the social and biological specificity of both sexes must be recognized and adjustments made to the ways questions are asked, hypotheses are generated, research subjects are chosen, and data are analysed. It is important to examine the definitions, concepts and methods used in research in occupational health to see whether they obstruct recognition of women's occupational health problems or interfere with gender-neutral standard setting. Unravelling the effects of sex on occupational health is complicated by the fact that women and men do not, by and large, work at the same jobs. Definitions of "work" must in some cases be adjusted to take account of women's occupations, just as definitions involving "health" must include women's biological specificity. Appropriate changes must be made to generate sex inclusive definitions of exposures to occupational hazards and of health effects. Methods for evaluating exposures typical of women's work must be developed. Women and their work must be appropriately included when standards for occupational exposures are set. If these adjustments are not made, women's occupational health problems will be seriously underestimated and understanding of health problems of both sexes will be hindered. Sociological analysis should also be done to reveal the mechanisms by which biased concepts and procedures develop and are reinforced. PMID- 8424184 TI - Reconceptualizing gender in physician-patient relationships. AB - In this paper, I propose that existing research on how women physicians relate to patients differently from men might benefit from a reconceptualization of gender. Toward this end, I present an ethnomethodological perspective on gender as an accomplishment and show how it contrasts with conventional perspectives on 'sex differences' and 'sex roles'. I review results of existing research on women and men physicians' relationships with their patients, highlighting the inadequacies of conventional perspectives to address this topic. Finally, I recast results of my own research on 'doctors' orders' in an effort to illustrate the utility of this perspective to research on gender's effects on the physician-patient relationship. PMID- 8424185 TI - Structure and struggle: implementing a social model of a well woman clinic in Glasgow. AB - Although the values of the women's health movement are widely shared in the U.K., they are little in evidence in the services providing health care for women. This article makes the distinction between a medical model of health care and a social model. It describes a research project which took place in which a well woman clinic was transformed from one which operated on a medical model to one which reflected those of a social model of health care. The article describes the problems which the project team faced in implementing the changes. PMID- 8424186 TI - Healthy bodies, social bodies: men's and women's concepts and practices of health in everyday life. AB - Using interview data from white, middle-class men and women, ages 35-55, the research explores the phenomenological, embodied aspects of health. Health is found to be grounded in a sense of self and a sense of body, both of which are tied to conceptions of past and future actions. Gender is a leitmotif. The body, as the focal point of self-construction as well as health construction, implicates gender in the everyday experience of health. The interplay between health, self, body, and gender at the individual level is linked to the creation of a sense of healthiness in the body politic of society. If social psychological theories of health are to reflect adequately the everyday experience of health, they must begin to take into account the body as individually and socially problematic. PMID- 8424187 TI - Keeping children healthy--the intermediate domain. AB - This paper draws on two recent research studies to consider negotiations and relationships between parents and health staff as regards child health care and child rearing, to which each side contributes. The value of the concept of an intermediate domain, located between the public world of paid work and the private world of the family is explored to throw light on the character of these negotiations and relationships. The implications of gender for relationships between parent and health staff are considered. PMID- 8424188 TI - Issues of gender in gamete donation. AB - Gamete donation refers to the practice whereby either semen or eggs are donated by a third party to enable infertile individuals or couples to become parents. This paper examines the way in which gender is deployed as a resource for organizing the meanings attached to that practice. The gender aspects of gamete donation are not always immediately apparent since semen and egg donation are often described as being essentially the same. However, a closer examination indicates that behind the claim of equivalence lies a set of unstated assumptions about their difference. These assumptions are tied to ideas about the ways in which men and women are thought to behave more generally in relation to reproduction and the family. This paper draws on two sources of empirical data to reveal how these assumptions are used: first, data from a detailed analysis of the Warnock Report (established by the British Government in 1982 to inquire into and make recommendations on techniques of human fertilization and embryology), which includes a cross-national and historical comparison with other government reports; second, data from a series of in-depth interviews with members of the Warnock Committee. The analysis of the reports suggests that historically semen donation was associated with 'deviant' sexuality (masturbation, adultery, illegitimacy) though paradoxically the extant nature of semen donation was then used to justify the acceptance of egg donation in later reports. This is despite the fact that in these later reports there are clear, albeit implicit, distinctions drawn between the two procedures in terms of donor motivation, the risks of being a donor, and the consequences of donation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424189 TI - New developments in south African health and safety legislation. PMID- 8424190 TI - Suction v. conventional curettage in incomplete abortion. A randomised controlled trial. AB - This randomised controlled trial of 357 patients who had had an incomplete abortion compared suction curettage with conventional curettage for evacuation of the uterus. The 179 patients undergoing suction curettage had a significantly lower intra-operative blood loss (P < 0.0001) and a significantly higher mean haemoglobin level at follow-up compared with the 178 patients who had conventional curettage. Suction curettage was a faster procedure and less painful. No difference was found between the two groups with regard to the incidence of post-abortal sepsis, or the re-evacuation rate. No problems were encountered with the use of suction curettage in the presence of uterine sepsis. In an era where blood transfusions should be kept to an absolute minimum, suction curettage will help to save blood in several ways. PMID- 8424191 TI - The potential benefit of pre-operative assessment of amputation wound healing potential in peripheral vascular disease. AB - Choosing the most distal amputation level that will heal is difficult in patients with peripheral vascular disease. From 1984 to 1988, 965 patients underwent 1,563 amputations for lower limb peripheral vascular disease at King Edward VIII Hospital, Durban. The primary amputation revision rate was 51% with a mortality rate of 23.1%. Random pre-operative assessment of amputation wound healing potential using a transcutaneous oxygen pressure index was investigated over the 4-year period, 1987-1990. This was responsible for a reduction in the amputation revision rate to 8.2% in patients tested. PMID- 8424192 TI - Peyronie's disease--a perspective on the disease and the long-term results of radiotherapy. AB - From 1966 to 1988, 98 of 108 patients with symptomatic Peyronie's disease received radiotherapy at our institution. In 11 of 61 patients (18%) who attended the clinic regularly for follow-up for longer than a year, new lesions distinct from the original lesions developed. This confirms that there is progression of the disease in a substantial number of cases after treatment. Long-term follow-up over an average of 111.5 months was achieved by means of a questionnaire in 47 of the 98 cases (48%). Forty-one of these patients (87.2%) had sexual intercourse after radiation. Twenty-eight of the 41 (68.3%) still have intercourse. Their average age at present is 59.6 years while the average age of the 13 patients (31.7%) not having intercourse is 70.9 years. The decline in sexual activity is thus age-related. Twenty-one of 25 patients (84%) experienced relief from pain, and angulation of the penis improved in 17 of 44 patients (38.6%) after radiotherapy. Radiotherapy may therefore be of benefit to patients with active Peyronie's disease and should be investigated in a randomised controlled study. PMID- 8424193 TI - Coronary artery disease and insulin resistance in the South African Indian. PMID- 8424194 TI - Q fever in Zimbabwe. A review of the disease and the results of a serosurvey of humans, cattle, goats and dogs. AB - Sera from 494 humans, 180 cattle, 180 goats and 27 dogs, collected from different regions of Zimbabwe, were examined by indirect fluorescence for antibodies reactive with phase II Coxiella burnetii antigen. Overall, 37% of humans were reactive at a titre of 1/40 or greater, and there was no evidence of age- or sex related differences in seroprevalence. A review of clinical and epidemiological features of Q fever is presented in order to alert health workers to this infection, which apparently occurs frequently in Zimbabwe even though clinical cases have not been reported. In animals, serological evidence of Q fever infection was found in 39% of cattle, but only 15% of dogs and 10% of goats. These results suggest that cattle are important reservoirs of C. burnetii in Zimbabwe. PMID- 8424195 TI - Robust and inexpensive equipment design for polymerase chain reaction detection of sequence mutations. Cystic fibrosis in a mother and 2 children analysed. AB - Every polymerase chain reaction (PCR) requires use of a temperature cycler for about 3 hours. Since there are many diagnostic tests using this technology, it is important that robust but inexpensive machinery is available. Such a stand-alone machine has been designed and used to analyse an interesting family in which a mother and her 2 children were diagnosed as having cystic fibrosis. PMID- 8424196 TI - The way ahead for medicine with the genomic code. PMID- 8424197 TI - Overload proteinuria. PMID- 8424198 TI - The labour ward analgesic service at King Edward VIII Hospital, Durban. AB - The provision of analgesic services to the labour ward at King Edward VIII Hospital was studied during a 1-week period. Of 249 patients, 113 (45%) received no analgesia whatsoever. Intramuscular pethidine was the commonest form of analgesia and was used in 97 patients (39%). Thirty-six patients (14%) received epidural analgesia and only 4 inhalational analgesia using nitrous oxide and oxygen (Entonox). A significant proportion of patients who received pethidine were given the drug between 1 and 3 hours before delivery, increasing the potential for opiate-related neonatal depression. Of the patients given opiate analgesia, 22 (23%) proceeded to caesarean section and could have been at increased risk of aspiration of gastric contents owing to delayed gastric emptying caused by the opiate. One hundred and eleven mothers (76%) who had an obstetric indication for epidural analgesia were denied it because of lack of medical staffing. PMID- 8424199 TI - The antenatal prevention of congenital syphilis in a peri-urban settlement. AB - The obstetric records of patients from Khayelitsha were examined to assess the efficiency of a system for the antenatal prevention of congenital syphilis, and to identify points of breakdown in the process. Seventy-seven (12.7%) of 607 mothers had serological evidence of syphilis, including 10 (32.3%) of 31 mothers who had received no antenatal care. Of 70 patients who required routine management, only 36 (51.4%) received 3 or more of the recommended 4 penicillin injections. Two main weaknesses in the system were identified. One was the centralisation of serological testing. This delayed results reaching the relevant unit, and was responsible for a high cumulative attrition of patients during the many stages necessitated by the centralised testing. The other was a 24.5% attrition of patients referred from the antenatal clinic to a separate sexually transmitted diseases clinic. PMID- 8424200 TI - HIV infection in Bophuthatswana. Epidemiological surveillance 1987-1989. AB - To assess the degree of HIV endemicity and the extent of its penetration into the general population in Bophuthatswana, a sentinel network of all the 11 hospitals and 3 prisons in the country was established in 1987. In addition, a population seroprevalence study was conducted in 1989. Between 5 January 1987 and 31 December 1989, serum specimens from 19,941 persons of all ages were tested for HIV. Overall, 34 subjects were found to be seropositive (prevalence 0.17%). Between 1 April and 31 August 1989, an HIV seroprevalence survey in a stratified multistage random sample of the resident population aged 15 years or older was also conducted. In this study, 2,111 persons were sampled but HIV antibody tests were done on 1553 only. A total of 12 serum specimens (0.8%) were reactive for anti-HIV by both Abbott EIA and Serodia HIV kits. However, in only 2 of these 12 sera was Western blot analysis positive. The overall prevalence of HIV infection in this study was 2/1553 participants (0.13%). These results demonstrate not only that HIV infection and the potential for its transmission are present in the Republic of Bophuthatswana but that they call for energetic and well-targeted control programmes. PMID- 8424201 TI - Population control and human development. PMID- 8424202 TI - Prevention of sexually transmitted disease. The Shurugwi sex-workers project. AB - Sex-workers play an important role in the spread of sexually transmitted diseases (STDs) and this article tries to show that they can also play an important role in their prevention. Community participation by sex-workers in the prevention of STDs can also decrease the incidence thereof. PMID- 8424203 TI - Inoculation of peritoneal dialysate fluid into blood culture bottles improves culture rates. AB - The aim of the study was to determine if direct inoculation of peritoneal fluid into Bactec blood culture bottles would improve the positive bacteriological yield compared with conventional techniques in continuous ambulatory peritoneal dialysis (CAPD) patients with peritonitis. All patients presenting with suspected peritonitis had peritoneal fluid injected directly into aerobic and anaerobic Bactec blood culture bottles as well as into sterile culture tubes. Thirty-seven paired samples were analysed. Twenty conventional cultures (54%) were positive compared with 33 (89%) done according to the Bactec system (P < 0.002). In only 1 case did the former technique prove superior. Direct inoculation of peritoneal fluid into Bactec blood culture bottles is therefore superior to conventional methods and has obvious therapeutic implications. PMID- 8424204 TI - Pregnancy outcome after in vitro fertilisation at the Pretoria unit. AB - This retrospective study was conducted to ascertain the success rate of a university-based in vitro fertilisation (IVF) programme. Over a 4-year period a total of 881 patients was aspirated with an 81.8% embryo transfer (ET) rate. This resulted in 150 biochemical pregnancies (20.8% per ET), and 92 births that produced 100 babies were recorded (12.8% per ET). Multiple pregnancies accounted for 8.7% of births and 6.5% of the 100 babies presented with minor congenital abnormalities. Delivery took place by caesarean section in 46% of cases. Birth mass of babies ranged from 1.06 to 4 kg with a mean of 2.944 +/- 0.629 kg. Mean gestational age was 260 +/- 18.2 days. Twenty-three percent of babies born were preterm, 13.8% of these were twins and presented with a low birth mass. It was concluded that the conception rate of patients did not differ with regard to the number of IVF attempts but that those who conceived during the first two attempts had a significantly better chance (P < 0.05) of carrying to term. PMID- 8424205 TI - What is normal? AB - The distinction between normality and abnormality forms the basis of medical practice. However, these words appear to have no more precise meanings in medicine than they do in conversational English. At least five contradictory definitions are described in the literature, and are in simultaneous use in everyday practice. The apparently arbitrary manner in which these definitions are chosen to evaluate different phenomena effectively means that medicine operates without a definition: certain phenomena are normal (by decree, as it were), and others are not. Actions based on such an arbitrary system are, of necessity, haphazard. The adoption of a precise, rigorously acultural definition of normality would unquestionably admit medicine to full membership of the family of objective sciences. PMID- 8424206 TI - Community participation in health service institutions. PMID- 8424207 TI - Exercise and the gastro-intestinal tract. AB - Approximately 50% of athletes will develop gastrointestinal symptoms at some stage in their careers. These range in severity from heartburn to gastro intestinal bleeding. Fortunately symptoms are usually mild and inconvenient, but in certain individuals they can be incapacitating. It is important to exclude the more common gastro-intestinal conditions before diagnosing exercise-related syndromes. However, once such a diagnosis has been made, therapeutic options are limited. The physiological role of the gastro-intestinal tract in fluid and energy replacement is increasingly being recognised. Without adequate replacements, performance may be limited. The volume of fluid ingested during endurance events needs to be limited to actual requirements; 500 ml/h is the average. Greater volume intake may be associated with overhydration and hyponatraemia. Glucose supplementation is essential for adequate performance in events of 2-3 hours' duration or longer. Studies of hyperosmolar carbohydrate solutions and their influence on energy and fluid emptying from the stomach suggest that higher carbohydrate concentration solutions than those often used by athletes may be advantageous. PMID- 8424208 TI - Notifiability of HIV and AIDS. PMID- 8424209 TI - A Pandora's box of HIV testing. PMID- 8424210 TI - Sensitivity of hospital budgets to changes in the relative weighting of outpatient department visits. PMID- 8424211 TI - SI units--can this use be justified. PMID- 8424212 TI - Vitamin A for measles. PMID- 8424213 TI - Lack of dual infection with HIV-1 and HTLV-I. PMID- 8424214 TI - Gonococcal infection in acute haemorrhagic conjunctivitis. PMID- 8424215 TI - Occupational lung disease. PMID- 8424216 TI - Ascorbic acid causes spuriously low blood glucose measurements. PMID- 8424217 TI - Intravascular intra-uterine transfusions for severe fetal iso-immunisation--a new technique in South Africa. PMID- 8424218 TI - [Another aspiration danger during anesthesia induction]. PMID- 8424219 TI - Opiate intoxication secondary to the smoking of crude opiates. PMID- 8424220 TI - Holoprosencephaly--the use of magnetic resonance imaging and application in antenatal diagnosis. PMID- 8424221 TI - The reversibility of cancer, 10 years on. PMID- 8424222 TI - A study of immunoglobulin G subclass levels in black and white patients with various forms of obstructive lung disease. AB - The association of various respiratory disorders with disturbances in immunoglobulin G (IgG) subclass levels is increasingly being recognised. This was a prospective study of the IgG subclass levels in 71 patients (37 white, 34 black) with various respiratory disorders associated with obstructive airways disease. Ten white patients with adult cystic fibrosis were studied, 4 of whom were colonised with Pseudomonas aeruginosa. Alterations in individual subclass levels were seen in these patients and abnormalities noted included a decrease in IgG3 and/or an increase in IgG1 and/or IgG2 levels. Of the 17 black and 16 white patients with asthma, 2 had absent IgG4 levels associated with severe and recurrent respiratory infections, 2 had deficient IgG3 and 1 decreased IgG1 levels. Several patients had increased levels of several IgG subclasses, of which IgG1 was the most commonly affected. Both atopy and recurrent chest infections occurred most often in the latter group of patients. In the studies of 17 black and 11 white patients with bronchiectasis, all but 3 white patients were shown to have some alteration in IgG subclass levels. The commonest deficiencies were an absence of IgG4 (3 cases), and an absence of all subclasses (2 cases). One of the latter patients had an associated deficiency of total IgG and IgM, and the other demonstrated pan-hypogammaglobulinaemia. Abnormalities of IgG subclass levels appear to be commonly associated with several respiratory disorders including recurrent infections, atopy and bronchiectasis. PMID- 8424223 TI - Back to the basics in laparoscopic cholecystectomy. PMID- 8424224 TI - Laparoscopic insufflation of the abdomen depresses cardiopulmonary function. AB - Recently we have used the laparoscope to remove the gallbladder in critically ill patients in order to spare them the operative trauma of laparotomy. However, increased intraperitoneal pressure may have deleterious cardiopulmonary effects. This was investigated in a dog model. Insufflation of the abdomen with carbon dioxide decreased the mean cardiac output to less than 80% of baseline (P < 0.004). This was aggravated by the reverse Trendelenburg position and partially alleviated by the Trendelenburg position. Mean arterial PCO2 and mean peak airway pressure significantly rose. These effects are of doubtful clinical significance in the majority of patients. However, to extend the benefits of laparoscopy to patients with decreased cardiopulmonary reserve, hemodynamic and carbon dioxide monitoring should be used because measures to improve venous return, augment cardiac output, and counteract the increase in PCO2 and peak airway pressure may be required. PMID- 8424225 TI - The impact of laparoscopic cholecystectomy on the operative experience of surgical residents. AB - The impact of laparoscopic cholecystectomy (LC) on the operative experience of surgical residents was assessed in a series of 787 cholecystectomies. During an initial 18-month period, residents participated in LC as operating surgeon and as first assistant or camera operator in 33% and 97% of cases, respectively. Operative time, cholangiography rate, conversion rate, and complications were not adversely affected by resident operators. Residents performed 87% of concurrent planned open cholecystectomies (OC). In comparison to the 6 months preceding LC: (1) The mean number of resident OCs decreased significantly while the total number of resident cholecystectomies was unchanged; (2) the proportion of OCs performed by PGY5 residents significantly increased at the expense of junior resident cases. LC can be safely integrated into surgical resident training by standard methods as for open procedures. Although resident operative experience has been redistributed, initial experience does not suggest that qualification in open biliary surgery has been compromised. PMID- 8424226 TI - Preoperative ERCP and intraoperative cholangiography in the age of laparoscopic cholecystectomy. PMID- 8424227 TI - Hernioscopic stuffing of direct inguinal hernia in female patients using resorbable mesh. AB - Since 1989 we have performed 21 endoscopic hernia repairs in 19 female patients. One recurrent hernia occurred 3 months after laparoscopic preperitoneal patch repair using a single layer of resorbable mesh. Hernioscopy was developed as the transcutaneous endoscopic CO2-gas dissection and subsequent inspection of the preperitoneal hernial sac. Hernioscopic stuffing of the preperitoneal hernial sac using resorbable patch material was performed in seven direct inguinal hernias and in one femoral hernia. Postoperative pain was minimal and convalescence was short. No recurrent hernia occurred during a 1-9-month follow-up. PMID- 8424228 TI - Laparoscopic repair of inguinal hernias using a totally extraperitoneal prosthetic approach. AB - This report describes a laparoscopic procedure for prosthetic repair of inguinal hernias using an extraperitoneal approach. A total of 51 primary direct and indirect hernias were repaired in this series, including 11 recurrent and 12 bilateral hernias. Operative time for this laparoscopic procedure was similar to that of the comparable open surgery and no unusual complications were noted. All patients were discharged the day following surgery and returned to work within 7 days. PMID- 8424229 TI - Laparoscopic assisted proctosigmoidectomy with extracorporeal transanal anastomosis. A pilot study. AB - Laparoscopic colon resections have often required an abdominal incision to remove the specimen and perform the anastomosis. Our aim was to mobilize the left colon and rectum using the laparoscope and perform a perineal proctosigmoidectomy with a primary end-to-end anastomosis. In eight pigs we used the operating laparoscope to mobilize the left colon, to ligate the inferior mesenteric artery at its origin, to ligate the inferior mesenteric vein as it crossed the left colic artery, and to fully mobilize the rectum. The rectum and sigmoid colon were then prolapsed through the anal canal, transected, and anastomosis was performed using an EEA stapler. The anastomosis was tested for structural and vascular integrity. Following the procedure, laparotomy was performed to estimate blood loss, to record visceral injury, and to examine the specimen for extent of resection. We were able to perform the resection and anastomosis in all animals with minimal blood loss and with high ligation of the vascular pedicle. There were no major visceral injuries. All anastomoses were perfused, patent, and intact. We concluded that when using the laparoscope in the porcine model, a low anterior resection and anastomosis can be performed safely with an adequate specimen without a laparotomy incision. PMID- 8424230 TI - ERCP, cholangiography, and laparoscopic cholecystectomy. The Society of American Gastrointestinal Endoscopic Surgeons (SAGES) opinion survey. AB - With the increased use of laparoscopic cholecystectomy (LC), the roles of preoperative ERCP and intraoperative cholangiography (IOC) may be changing. SAGES members were surveyed to define opinions regarding use of ERCP and cholangiography during LC. Thirty-seven percent of the surveys were returned. Most respondents (83%) performed LC, reporting data on 19,747 LCs. Conversion to open cholecystectomy was required in 4% of cases. Complications were reported in 1.7% patients. IOC was attempted in 51.2% cases and was successful in 73%. Routine IOC was only recommended by approximately 50% of respondents. However, 80% recommended IOC for patients with multiple small gallstones and a dilated cystic duct. If preoperative liver function tests (LFTs) were mildly elevated (1.5 x normal), only 56% of respondents recommended preoperative ERCP. However, 73% of respondents suggested preoperative ERCP for more severe LFT abnormalities. If preoperative ERCP demonstrated choledocholithiasis, most (85%) recommended endoscopic clearance of stones followed by LC. These opinions will be helpful in establishing practice standards for LC. PMID- 8424231 TI - The reliability of cecal landmarks during colonoscopy. AB - Confirming colonoscopic intubation of the cecum can be a laborious, time consuming, and often frustrating endeavor. Anatomic landmarks may offer visual clues of cecal intubation, but the predictability of this evidence is unclear. The presence of three cecal landmarks, alone and in combination, were evaluated to precisely define their reliability. Between February and October of 1991, 601 of 708 (85%) consecutive colonoscopic examinations were able to be completed to the cecum as confirmed by fluoroscopy. All three cecal landmarks studied were present in 64% (386/601), two cecal landmarks in 32% (189/601), and one cecal landmark in 4% (26/601) of the patients. Therefore, at least two cecal landmarks were identified in 96% (575/601) of the patients. The ileocecal sphincter was identified in 98% (591/601) of patients overall, in 98% (185/189) of patients and 2 cecal landmarks, and in 77% (20/26) of patients with 1 cecal landmark. The appendiceal orifice was seen in 87% (524/601) of patients overall and in 72% (137/189) of patients with 2 cecal landmarks. Transillumination through the abdominal wall was possible in 74% (447/601) of patients overall and in 30% (56/189) of patients with 2 cecal landmarks. In summary, the ileocecal sphincter is the most reliable cecal landmark and is invariably visualized, even when all other landmarks are obscure. While other cecal landmarks, such as the appendiceal orifice and transillumination, are consistently identified, they are most valuable when found in association with the ileocecal sphincter. PMID- 8424233 TI - Laparoscopic hysterectomy. A preliminary study. AB - Thirty-three patients were selected for laparoscopic hysterectomy and operated on in the Department of Obstetrics, Gynecology and Reproductive Medicine of Clermont Ferrand University Hospital. Surgical techniques included blunt dissection with scissors and bipolar coagulation to achieve hemostasis. A case was considered successful when all the uterine vessels were treated by laparoscopy. Twenty-four cases were completed laparoscopically (72.7%). None of these patients had postoperative bleeding; 22 had an uneventful postoperative recovery. Nine procedures were converted to laparotomy (27.3%), five because of a difficult or unsatisfactory hemostasis. We conclude that in selected cases, a total hysterectomy can be performed safely by experienced laparoscopists. Further technological progress is necessary to make this procedure more acceptable. Its value as compared to the others will have to be demonstrated. PMID- 8424232 TI - Endoscopic clearance of the pancreatic duct in chronic pancreatitis with severe pain. AB - In a phase I study endoscopic removal of pancreatic duct stones and protein plugs was attempted in five patients suffering from chronic pancreatitis with severe chronic pain. The pancreatic duct contents could be extracted after successful sphincterotomy in three patients. Clearance of the pancreatic duct was followed by complete or partial relief of pain. The follow-up period was 17-48 months. Endoscopic extraction is, however, not without complications; it is technically difficult, and many attempts may be required. The combination of endoscopic therapy and extracorporeal shock-wave lithotripsy may be a better alternative. PMID- 8424234 TI - Arthroscopic repair of traumatic longitudinal meniscal tears. A 3 to 5-year follow-up. AB - The healing potential of the meniscal tissue has been known for a century but has only been broadly introduced into surgical treatment during the last years. Open surgical suture of the meniscus has increasingly been replaced by arthroscopic refixation. We report 34 meniscal refixations with a minimal follow-up of at least 3 years and a mean of 4 years. Using our own simple and economic surgical technique, 34 refixations were performed in 32 patients from January 1987 to December 1988. All patients had traumatic meniscal tears close to the capsule. Frequently the injury was associated with a fresh or old rupture of the anterior cruciate ligament. Without additional trauma, one meniscus had to be partially resected after 4 months; a second one was partially resected in an unstable knee. Both the clinical examination and the satisfaction of the patient demonstrate that meniscal refixation is feasible and appropriate with a correct indication. PMID- 8424235 TI - Thoracoscopic ligation of the thoracic duct. AB - Traditional operative management for chylous drainage refractory to conservative therapy is thoracic duct ligation via right open thoracotomy. This case report details successful thoracoscopic ligation of the thoracic duct for a chylous leak following a left neck dissection. Since the thoracoscopic approach is less morbid than open thoracotomy, early operative management is recommended for thoracic duct injuries. PMID- 8424236 TI - Laparoscopic cholecystectomy: an approach without pneumoperitoneum. AB - Diagnostic pneumoperitoneum, which has been considered the first step of any laparoscopic procedure, is no longer an absolute necessity. We devised an alternative to pneumoperitoneum or abdominal insufflation by upward and outward traction on the anterior abdominal wall with a "hanger lifting method" using subcutaneous wiring. Fairly good room was produced intraabdominally, which was enough in which to perform the cholecystectomy procedure. We have successfully performed 40 cases of laparoscopic cholecystectomy with this procedure. No complication was experienced with this method and, moreover, excess instrumentation and complications related to pneumoperitoneum were avoided. PMID- 8424237 TI - Abdominal wall lift. Low-pressure pneumoperitoneum laparoscopic surgery. AB - A method of abdominal wall lift has been developed and evaluated clinically in this unit during the past 18 months. It permits the conduct of laparoscopic procedures at an intraabdominal pressure of 6-8 mm Hg. The technique was introduced for laparoscopic surgery in patients with preexisting cardiac disease and chronic bronchitis. The procedure, by lifting both the abdominal wall and the falciform ligament together, also elevates the central portion of the liver (segments 3-5), thereby improving the surgical exposure. For this reason it is now also used in fit patients with ptotic livers or hypertrophied quadrate lobes undergoing laparoscopic cholecystectomy and common bile duct exploration, and to facilitate left subhepatic exposure in patients during laparoscopic antireflux surgery and vagotomy. PMID- 8424238 TI - Laparoscopic gastrostomy using T-fasteners as retractors and anchors. AB - Most feeding or venting gastrostomies can be placed percutaneously via gastroscopy. Laparotomy is required if gastroscopy is not possible--for example, in patients with esophageal strictures or large tumors. We have developed a new technique of laparoscopic gastrostomy, and used it successfully in three patients. The key to this technique is the T-fastener, a nylon suture attached to a metal T-bar, which is introduced via a slotted needle percutaneously and dislodged inside the stomach lumen. Four T-fasteners secure the stomach wall to the anterior abdomen. A balloon catheter gastrostomy tube is then placed over a J wire through the center of these T-fasteners. This technique requires no gastroscopy or intracorporeal suturing and needs only one port for the laparoscope. It is safe and simple and can be used for patients who require a gastrostomy in whom gastroscopy is not possible or is risky. PMID- 8424239 TI - The use of ERCP in the management of common bile duct stones in patients undergoing laparoscopic cholecystectomy. AB - The purpose of this study was to evaluate the indications and results of endoscopic retrograde cholangiopancreatography (ERCP) for gallstone disease since the advent of laparoscopic cholecystectomy. In our personal series of 410 consecutive cases of laparoscopic cholecystectomy, we found 17 common bile duct (CBD) stones; seven were identified by preop ERCP, nine at laparoscopy by intraoperative cholangiography, and one postop by ERCP. We have performed preop ERCP in 21 patients (5.1%); CBD stones were found in seven. Our indications for preop ERCP were elevated liver function tests, dilatation of the common duct by ultrasound, or a history of jaundice/pancreatitis, and all stones were successfully removed by endoscopic sphincterotomy. At laparoscopic cholecystectomy nine patients were found to have stones; one was treated with laparoscopic methods, four with open CBD exploration, and four by postop endoscopic sphinecterotomy. Post-laparoscopic cholecystectomy, five patients underwent ERCP for pain or increased liver function tests suggestive of common duct stones. One of the five was found to have stones and these were successfully removed by endoscopic sphincterotomy. ERCP is very useful as a diagnostic and therapeutic modality in laparoscopic cholecystectomy patients with suspected CBD stones. Elevated liver function tests and dilated CBD by ultrasound are the most accurate predictors of stones. Endoscopic sphincterotomy is a more effective route, at present, for stone removal than a laparoscopic approach. PMID- 8424240 TI - [Do we need billions for hospitals?]. PMID- 8424241 TI - [Behavioral modification in medical practice]. PMID- 8424242 TI - [Surgery for myopia]. PMID- 8424243 TI - [Changing blood bank service. Delayed thanks to Jehovah's Witnesses]. PMID- 8424244 TI - [Production and consumption of blood and blood products in Norway. Factors of quality assurance in transfusion medicine]. AB - In Norway self-sufficiency has been achieved with regard to all major blood products, despite a relatively low number of blood donations. There is good balance in the consumption of the various products. The total consumption of erythrocytes has gone down only slightly since pre-HIV times, while consumption of thrombocytes has increased. There is probably an over consumption of erythrocytes, albumin and plasma, and more attention should be given to the consistent application of principles of modern haemotherapy. If each department is given responsibility for its own economy, improvement may be achieved in this respect. PMID- 8424245 TI - [Exposure to blood among hospital physicians]. AB - We used a questionnaire to study the working situation of physicians in two large Norwegian hospitals. During the last six months 40% of the doctors reported one or more episodes of exposure to blood when transfer of infectious agents such as hepatitis B-virus or HIV would have been possible. The fact that so many doctors had been exposed to blood conflicts with the low number reported to the hospital department of the occupational health service. Surprisingly, 40% of the physicians who reported exposure were physicians not taking part in surgical procedures. All hospitals should establish follow-up procedures for employees exposed to blood or other possible infectious agents, and physicians should be urged to follow these routines. PMID- 8424246 TI - [Graduates from Tromso--do they stay in Northern Norway? Geographic origin determines selection of professional practice location in Northern Norway]. AB - The impact of the Medical School, University of Tromso on the distribution of physicians in rural areas in North Norway was evaluated by means of a mailed questionnaire. The survey covered eleven graduation classes (417 physicians), and the response rate was 84.2%. The establishment of a new medical school in North Norway has clearly had beneficial effects: A total of 56.1% of the graduates stay on as doctors in these areas. Among those who also grew up in the northern parts of Norway, the proportion is 82.0%, compared with graduates who grew up in the southern parts where the proportion is 37.7%. The results clearly demonstrate that one of the main objectives of the Medical School at the University of Tromso, to educate physicians who prefer to work in these areas, has been achieved. PMID- 8424247 TI - [Elective periods in the medical curriculum. Madness or academic challenge?]. AB - The medical curriculum at the University of Tromso reserves 23 weeks, divided into four periods, for elective work, including 12 weeks at the end of the fifth year, when the student carries out an independent study and writes a short thesis. 84.2% of 417 physicians (graduation year 1979-89) answered a mailed questionnaire asking them to evaluate these periods. The postgraduates report high levels of satisfaction with this part of the curriculum, thought they had benefited from it. As many as 86.0% evaluate the gain from the first three periods as good or very good. 26.8% started working on their thesis before the final 12 week period. 88.1% found the fourth, and longest period, assigned for their thesis to be an important part of the curriculum. One third had also published their material, half of these in international journals. 75.6% found the skills acquired during the final elective period to be useful in their present work, independent of specialty. PMID- 8424248 TI - [Evaluations and examinations at the Tromso medical school. Evaluation by students after graduation]. AB - The medical curriculum at the University of Tromso is based on an organ-system model with full integration of the three main disciplines (basic science, clinical and community medicine). There are only three examinations during the six-year course. We present the results of a survey among the first 417 physicians educated at the Medical School in Tromso. A mailed questionnaire was answered by 84.2%. More than 80% stated that it was of great importance to evaluate the form and contents of the lectures and their supervisors' teaching abilities. More than 96% found that integration of the examinations was achieved in accordance with the intention. Furthermore, the examinations were also relevant for them as physicians. PMID- 8424249 TI - [Acute and chronic effects of deep diving on the nervous system]. AB - Diving deeper than 180 metres of seawater (msw) will impose neurological symptoms in most divers. Atactic signs and abnormal EEGs were found in five of 18 divers immediately after deep diving. Neuropsychological testing before and after deep diving in 64 divers revealed a reduction in autonomic reactivity (48%), increased hand tremor (27%) and impairment of spatial memory and reduced finger coordination (8%) after the dives. These results had not improved one year later. A follow-up study of 40 divers one to seven years after their last deep dive revealed that the divers experienced more problems of concentration and paresthesia in feet and hands than the controls. Two had had seizures, one had suffered episodes of transitory cerebral ischemia and one had experienced transitory global amnesia after the deep dives. In the future, oil installations at depths below 180 msw should be installed and maintained with remote control and robot technology. PMID- 8424250 TI - [Control of scabies in the district of Ytre Nordhordland. An experiment within community medicine in the 1860's]. AB - 19th century Norway may be likened to the developing countries of today. Scabies was very prevalent, and traditional approaches to treatment were in vain. The poor peasant population would not spend money on such. Thomas Collett (1835-98), the local doctor in a rural district of western Norway, initiated a programme for community control of scabies. The local Board of Health agreed to purchase a large amount of sulphur ointment. 1/8 of a barrel of ointment was placed at the disposal of local teachers. The teachers were instructed to initiate treatment of school-children who were found to be infected with scabies. The household to which the child belonged was also to be treated, as well as the other children at the school. Gradually, the prevalence of scabies declined. Self-care increased, and eventually, the peasants ended up making their own sulphur ointment. Experiences and recommendations from developing countries of today agree with the strategy adopted by Collett. Individual medical treatment of cases and their contacts is too costly, and fails to reduce the total prevalence of scabies. PMID- 8424251 TI - [Changing patients' health behavior--consultation and physician-patient relationship]. AB - Studies indicate that doctors' advice concerning changes in patients' lifestyle meet with relatively little success. The article gives a summary of empirical studies on patient compliance in relation to the doctor's behaviour and attitudes; the quality of the patient-doctor relationship; and the content and form of the doctor's message. A patient-oriented style of communication; an ability to explore and discuss patients' expectations and behaviour; a warm and friendly approach; and an ability to gain the trust of the patient, are attributes that have been found to be effective in influencing patients' behaviour. The content of the information should be limited and specific, and the delivery should be structured. Perhaps the most important factor is a shift in power and control from the doctor to the patient when encouraging changes in health-related behaviour. PMID- 8424252 TI - [Theory of behavior and preventive health care in medical practice]. AB - A number of theories and models of special relevance to individual health education are presented, such as: the Knowledge-Attitude-Practice model; Social Learning Theory and Health Locus of Control; the Health Belief Model; the Theory of Reasoned Action; and Bandura's Social Cognitive Theory. Recently, promising attempts have been made to integrate several of the models. Across the models, the most important regulatory factors for behaviour seem to be social norms, personal expectations and environmental reinforcements. These factors should be considered by the doctor in attempts to induce changes in patients' health related behaviour. In the future development of health behaviour models, social and environmental factors will probably be given even more attention. PMID- 8424253 TI - [Demographic factors and pregnancy--who chooses abortion?]. AB - The number of pregnancies terminated as induced abortions has been analysed from data recorded in the Norwegian Birth Registry and the Norwegian Abortion File at the Central Bureau of Statistics. During the time period 1 January 1979 to 31 December 1991, 871,439 pregnant women were registered. Among single women the proportion of pregnancies terminated as induced abortions varied between 40 and 75% throughout the study period. Cohabitant women chose abortion significantly more often than married women. Among married women the proportion of pregnancies interrupted varied with the woman's age and the number of children. Among women 20-34 years of age with one or no child, more than 97% of the pregnancies proceeded to term. Women undergoing abortion comprise a heterogeneous group, judged by demographic factors. Future studies must focus on social factors as well as demographic factors, in order to uncover the complexity of the reasons for choosing abortion. PMID- 8424254 TI - [Changes in consumption of blood--the significance of resource related and economic factors]. AB - During the period 1985 to 1991 there has been a marked change in consumption of blood, and at the National Hospital the number of transfusions decreased by 52% from 41,747 units. However, the number of admitted patients annually has remained unchanged and the patient pattern includes more complicated cases. We conclude that cost accounting, consumer-adapted reporting, vigilant follow-up and an active transfusion committee are important factors for the changes in blood consumption. Furthermore, the professional attitude towards surgical blood conservation techniques and modern haemotherapy have been consolidated at the hospital concerned. PMID- 8424255 TI - [Pain and pain relief in newborn and older infants]. PMID- 8424256 TI - [Use of benzodiazepines in hospitals--a frustration for general practitioners]. PMID- 8424257 TI - [Centralization of obstetric care and perinatal mortality in Norwegian municipalities during 1986-90]. PMID- 8424258 TI - [Behavior is formed and selected by its consequences]. PMID- 8424259 TI - [The lobotomy debate in Norway]. PMID- 8424260 TI - [Torture in Israel]. PMID- 8424261 TI - [Prohibition of blood transfusions among Jehovah's Witnesses]. PMID- 8424262 TI - Cytokines in transfusion medicine. PMID- 8424263 TI - Transfusion transmission of human T-lymphotropic virus types I and II: serologic and polymerase chain reaction results in recipients identified through look-back investigations. AB - To determine the transmissibility of human T-lymphotropic virus types I and II (HTLV-I and HTLV-II) via transfusion, persons who, from 1983 to 1989, received blood components donated by persons who subsequently tested anti-HTLV-I-positive were evaluated. It was found that 16 (30%) of 54 evaluable recipients of transfused cellular components became infected with one of the HTLVs: 8 had HTLV I and 8 had HTLV-II. Forty percent of platelet recipients and 28 percent of red cell recipients acquired infection. The rate of transmission of HTLV-I and HTLV II was significantly correlated with storage age of red cell units prior to transfusion: 47 percent for red cells stored < or = 14 days and 0 for red cells stored > 14 days (p < 0.01). Multiple confirmatory serologic tests performed in 46 anti-HTLV-I enzyme immunoassay-negative recipients revealed that HTLV infection could not be excluded in 3 recipients of blood components from HTLV-II infected donors. Polymerase chain reaction established HTLV-II infection in one recipient, and the other two recipients could not be classified with respect to HTLV infection status. It appears that some HTLV-II-infected transfusion recipients will not be detected by existing HTLV-I antigen-based reagents. If it is deemed necessary to initiate or continue look-back programs to detect transfusion transmission of HTLV-II infection, it is suggested that the current testing algorithm be modified in selected cases. PMID- 8424265 TI - The routine use of soluble Lewis antigens in the identification of Lewis system antibodies. AB - Approximately 3 percent of the blood samples tested at the authors' laboratory have contained unexpected red cell (RBC) antibodies, of which about one-third have been anti-Le(a) or anti-Le(b). Although these antibodies are usually clinically insignificant, considerable workload was expended in their identification. In hopes of reducing this workload, an alternative approach for identifying these antibodies was implemented. If a patient had no history of unexpected antibodies, with the exception of Lewis system antibodies, and if the results of serum antibody detection testing were typical for the presence of anti Le(a) and/or anti-Le(b), the serum was treated with commercially standardized soluble human Le(a) and Le(b) blood group substances, and the Lewis phenotype of the patient's red cells was determined. If the treated serum demonstrated specific antibody neutralization and the red cell phenotype was Le(a-b-), the results were interpreted as positive for anti-Le(a) or anti-Le(b), and no further antibody identification workup was performed. If the patient was Le(a+b-) or Le(a b+), or if the treated serum failed to demonstrate specific antibody neutralization, the serum was tested against panels of reagent RBCs that were licensed for the identification of unexpected antibodies. During the study period (8-1-89 to 6-16-90), 498 of 46,965 patients' samples met the criteria for testing for specific antibody neutralization. Of these 498 blood samples, 452 showed specific neutralization of antibody activity by Lewis substance and were Le(a-b ). Only 1 of these samples was discovered to contain an additional non-Lewis antibody (anti-Bg).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424264 TI - Cytokine production in IgG-mediated red cell incompatibility. AB - The transfusion of incompatible red cells may result in fever and systemic symptoms. The mechanisms by which these symptoms are produced in the setting of antibodies that do not usually fix complement, as in the Rh system, are obscure. It has been hypothesized, on the basis of their known biologic activities, that a specific set of cytokines may be involved in such transfusion reactions. Therefore, the production of the inflammatory cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-8 (IL 8) by human monocytes in response to red cells sensitized with anti-D was investigated, as a model of IgG-dependent hemolytic transfusion reactions. IL-1 beta, IL-6, and IL-8 were detectable in the culture supernatants at 4 to 6 hours and increased up to 24 hours, whereas TNF peaked at 6 hours. Immunocytochemical stains of cell preparations demonstrated IL-1 beta, IL-8, and TNF in monocytes engaged in erythrophagocytosis. IL-8 production and phagocytosis could be inhibited by monomeric IgG, but Fab fragments of a monoclonal antibody specific for the low-affinity IgG receptor Fc gamma RII could not be, which suggests the involvement of the high-affinity receptor Fc gamma RI. Neutralizing antisera to IL-1 beta and TNF did not abrogate the production of IL-8, which suggests that sensitized red cells serve as a primary signal for this cytokine. These findings indicate that the production of inflammatory cytokines by phagocytes may be responsible for the symptomatology of IgG-mediated hemolytic transfusion reactions. PMID- 8424266 TI - Mechanisms of white cell reduction in red cell concentrates by filtration: the effect of the cellular composition of the red cell concentrates. AB - The effect of platelets on the removal of white cells (WBCs) from 16 to 24-hour old red cell (RBC) concentrates by filtration was studied. RBC concentrates with various concentrations of platelets and WBCs were filtered on a cellulose acetate column filter and on three polyester flatbed filters. The microscopic study revealed that lymphocytes and most monocytes were captured in the smaller pores of the fiber network, irrespective of the brand of filter, the type of filter material, or the prefiltration platelet amount in the RBC concentrates. In contrast, efficient granulocyte depletion depended on granulocyte-platelet interaction and on the filter material. In the presence of platelets, granulocytes were captured in the top part of the column filter or in the coarse layers of two of the flatbed filters, where platelets covered the fibers. Platelet depletion of the RBC concentrates prior to filtration diminished the contribution of these parts of the filters to granulocyte capture. A larger part of the column filter or the fine layers of the flatbed filters were now required for granulocyte capture. In one of the flatbed filters, granulocyte-platelet interaction occurred mainly in the fine layers, which ended in blockage of this filter after the filtration of variable volumes (250-600 mL) of standard RBC concentrates. A quantitative estimation of the effect of platelets on the WBC reduction capacity found that all three flatbed filters had a highly significant decrease (p = 0.001) in WBC-reduction capacity for platelet-depleted or buffy coat-depleted RBC concentrates, as compared with standard RBC concentrates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424267 TI - Use of erythropoietin to increase the volume of autologous blood donated by orthopedic patients. AB - For patients who donate blood for autologous use and undergo major orthopedic surgery, low basal hematocrit (Hct) is the major cause of allogeneic blood exposure. To determine whether recombinant human erythropoietin (rHuEPO) could increase autologous blood procurement and reduce allogeneic blood exposure, a prospective randomized study was conducted in 50 women undergoing total hip replacement who had basal Hct < 40 percent (0.40). Patients were randomly placed in three groups: those receiving placebo, those receiving 300 U of rHuEPO per kg, and those receiving 600 U of rHuEPO per kg every 3 to 4 days for 21 days. Oral iron (125-270 mg/day) was given; in the last 24 patients, 100 mg of iron saccharate was administered intravenously at each donation. At each visit, 350 mL of blood was collected if Hct was > or = 34 percent (0.34). Patients receiving rHuEPO donated a greater amount of blood for autologous use than did patients in the placebo group (4.5 +/- 1.1 vs. 2.8 +/- 0.6 units; p < 0.05) and received a significantly lower amount of allogeneic blood (1.2 +/- 1.4 vs. 0.4 +/- 0.8 units; p < 0.05). No difference between the effects of the two doses of rHuEPO was observed. Iron support was a critical factor in the efficacy of treatment. No untoward effects were observed. The rHuEPO emerged as a safe and effective treatment, with adequate iron support, by which to increase preoperative deposit of autologous blood and to reduce exposure to allogeneic blood for patients with low basal Hct. PMID- 8424268 TI - Use of a questionnaire to identify potential blood donors at risk for infection with Trypanosoma cruzi. AB - Trypanosoma cruzi is the protozoan parasite that causes American trypanosomiasis (Chagas' disease). Chagas' disease is endemic in Latin America. The infection is usually seen in poor people who live in rural areas in substandard housing, where they are bitten by infected reduviid bugs. Transmission also can occur by blood transfusion. Infected individuals who immigrate to the United States might donate blood if they are asymptomatic and unaware of their infection. This study evaluated the usefulness of a questionnaire for identifying T. cruzi-infected individuals among prospective blood donors who met all American Association of Blood Banks, Food and Drug Administration, and State of California criteria for donor eligibility. Seventy-two of 3492 otherwise eligible donors were disqualified because of their answers on the questionnaire. Forty-five of these 72 agreed to be tested serologically, and 2 were positive for T. cruzi antibodies. One of six autologous blood donors tested also was positive for T. cruzi antibodies. We conclude that the questionnaire selected a subgroup of Latin Americans at high risk for T. cruzi infection. The deferral of these high-risk individuals clearly reduced the risk of transmission of T. cruzi by transfusion, without intolerably decreasing the supply of donated blood. PMID- 8424269 TI - Report on the Fifth International Society of Blood Transfusion Platelet Serology Workshop. AB - The Fifth International Society of Blood Transfusion (ISBT) Platelet Serology Workshop took place in 1990. A total of 21 laboratories participated in the wet workshop. Results were discussed at a meeting of the International Committee on Standardization in Haematology (ICSH) Expert Panel on Platelet Serology at the Joint Congress of the ISBT and the American Association of Blood Banks in Los Angeles in November 1990. Each of the participants analyzed 37 serum samples for platelet-reactive alloantibodies and isoantibodies. In addition, two drug dependent (quinine) platelet-specific antibodies had to be identified among a subset of seven sera. For the first time in a platelet serology workshop, five platelet suspensions had to be typed for the alloantigens of the HPA-1, -3, -4, and -5 systems. PMID- 8424270 TI - Granulocyte-macrophage-colony-stimulating factor: a review from preclinical development to clinical application. PMID- 8424271 TI - Directed donation and the developing world. PMID- 8424272 TI - Neonatal immune response to red cell antigens. PMID- 8424273 TI - Safety of preoperative blood donation (for autotransfusion) for cardiac surgery patients. PMID- 8424274 TI - Preliminary publication of standards. PMID- 8424275 TI - High-pressure injection injuries of the hand. AB - High pressure injection trauma to the upper extremity is a rare but potentially limb-threatening injury. The index finger on the non-dominant hand of young male workers is most commonly involved. Injected materials include paint, grease and diesel oil. Prompt recognition and initiation of treatment is necessary if a disastrous outcome is to be avoided. Primary treatment consists of surgical decompression and debridement. Amputation may be necessary. With early aggressive treatment, satisfactory results can be obtained. The use of high-pressure devices in industry has resulted in an unusual but potentially limb-threatening injury to the upper extremity known as the "high-pressure injection injury." This injury is a surgical emergency that demands prompt diagnosis and treatment if a disastrous outcome is to be avoided. The purpose of this paper is to present our recent experience and review the clinical features, evaluation and treatment principles. PMID- 8424276 TI - Esthesioneuroblastoma: diagnosis, prognosis, and treatment. AB - Esthesioneuroblastoma is a rare neoplasm arising from the olfactory epithelium and characterized by slow growth, frequent local recurrence, and metastasis. This tumor has no known etiologic cause and has been studied from clinically retrospective analysis. The historic treatment of esthesioneuroblastoma has included surgical resection, adjuvant radiation therapy, and chemotherapy for advanced disease. We describe a case of esthesioneuroblastoma which illustrates the diagnostic, prognostic, and therapeutic aspects of this uncommon neoplasm. PMID- 8424277 TI - Lyme disease redux: the legacy of Sven Hellerstrom. PMID- 8424278 TI - Teen access to cigarettes in Green Bay, Wisconsin. PMID- 8424279 TI - Wisconsin steps forward as a national leader for reform. PMID- 8424280 TI - Propofol induces bronchodilation in mechanically ventilated chronic obstructive pulmonary disease (COPD) patients. AB - The aim of this study was to evaluate the effects of propofol administration (2 mg.kg-1 i.v.) on the airways resistances and respiratory mechanics of patients affected by COPD exacerbation, requiring mechanical ventilation. Twenty patients required anaesthesia for diagnostic or therapeutic procedures. Fourteen consecutive patients were divided at random into two groups: Group P received propofol and Group C (control) received only Intralipid 10%; an additional group of six patients received i.v. flunitrazepam (0.03 mg.kg-1). Lung mechanics (dynamic and static compliance, peak inspiratory pressure, intrinsic positive and expiratory pressure, minimal and maximal resistances of the respiratory system) were evaluated in basal conditions and 3 and 6 min after propofol, Intralipid or flunitrazepam administration. We did not observe significant variations of the evaluated variables after Intralipid or flunitrazepam (Groups C and F), while in patients who received propofol (Group P), we observed the following modifications: dynamic compliance increased from 2.3 +/- 0.3 to 2.8 +/- 0.4 ml.kPa-1 (P < 0.05), peak inspiratory pressure decreased from 3.3 +/- 0.7 to 2.8 +/- 0.4 kPa (P < 0.05), minimal resistances of the respiratory system (that mainly reflect airways resistances) decreased from 1 +/- 0.2 to 0.7 +/- 0.2 kPa.l 1 x s-1 (P < 0.01). Our results suggest that propofol induces bronchodilation in mechanically ventilated COPD patients, and that this effect is not related specifically to the induction of general anesthesia. PMID- 8424281 TI - Enhanced pneumonia resolution by inhalation of nitric oxide? AB - We present a patient with severe bacteraemic pneumococcal pneumonia associated with severe hypoxaemia, where nitric oxide (NO) 15-40 ppm was added to the inspired gas. Nitric oxide therapy improved gas exchange, reduced pulmonary vasoconstriction and peak airway pressure. The patient survived. We observed an unexpected rapid and complete disappearance of bilateral pulmonary infiltrates during the first 120 h of the 7-day NO inhalation period. PMID- 8424282 TI - Prolonged total extracorporeal lung assistance without systemic heparinization. AB - A 16-year-old female developed severe ARDS in her single remaining lung following pneumonectomy for blunt trauma. Total extracorporeal lung assist (ECLA) for 40 days using a covalently heparin-coated circuit proved lifesaving. Systemic heparinization was not applied, as the heparinized surface by itself prevented clotting of the extracorporeal circuit. Systemic primary fibrinolysis developed but was not associated with major bleeding. A veno-right ventricular cannulation technique was used and maximum venous drainage for the extracorporeal circulation was achieved by elevating the bed 50 cm from the floor. This allowed extracorporeal blood flow (ECBF) approaching cardiac output (CO) and complete extracorporeal replacement of lung function. After 40 days, lung recovery allowed discontinuation of ECLA. Five days later the patient suffered serious lung collapse and was operated for a bronchopleural fistula. The patient was extubated 4 weeks after terminating ECLA and discharged in good condition 5 weeks later. PMID- 8424283 TI - Comparison of two anesthesia techniques on perioperative insulin response to i.v. glucose infusion in children. AB - Perioperative blood glucose and insulin levels were measured in children (1-9 years of age) randomly assigned to two groups according to anesthesia technique, general anesthesia (group GA) or general anesthesia combined with regional anesthesia (group RA). Children in the GA group (n = 10) received halothane and opioids, while children of the RA group received epidural anesthesia with bupivacaine (0.25%) and adrenaline combined with halothane anesthesia (n = 10). Children in both groups received 2.5% dextrose in 0.4 N saline administered by volumetric infusion pumps throughout the study period, the infusion rate being adapted to the child's age. Blood samples for glucose and insulin determinations were obtained: at induction, at the end of surgery, and 30, 60 and 120 min after surgery. In response to an identical glucose load, blood glucose levels increased significantly in both groups (P < 0.001), while no differences between groups were observed. Insulin levels did not change significantly postoperatively in the GA group (P = 0.058), while a significant increase was observed in the RA group (P < 0.001). Insulin/blood glucose ratio increased significantly only in the RA group (P < 0.05). The higher insulin secretion in response to glucose infusion in the RA group compared to the GA group may indicate an increased peripheral insulin resistance after regional anesthesia or, more likely, this secretion may be beneficial in contributing to improve postoperative nitrogen balance. PMID- 8424284 TI - Slow potentials of EEG burst suppression pattern during anaesthesia. AB - EEG burst suppression pattern was recorded during propofol and isoflurane anaesthesia. Using a long time constant, i.e. a high-pass filter with low cut-off frequency, revealed slow potentials, DC shifts, during bursts. These have been extensively studied in conjunction with cognitive-evoked potentials and epileptic discharges, but not previously in burst suppression EEG. PMID- 8424285 TI - Modified laryngeal mask as an aid to fiberoptic endotracheal intubation. PMID- 8424286 TI - Flexible fibreoptic bronchoscopy via the laryngeal mask. AB - The efficacy of flexible fibreoptic bronchoscopy through the laryngeal mask was investigated in 20 patients under total intravenous anaesthesia with propofol, fentanyl, atropine and suxamethonium. Mask size 4 was used for men and size 3 for women. Ventilation was performed with oxygen in air, FIO2 0.6. The ventilatory pressures were median 18 (9-40) cmH2O (1.8 (0.9-3.9) kPa) before the bronchoscope was inserted. When the tip of the bronchoscope was above the vocal cords the ventilatory pressures increased to 22 (10-43) mmHg (2.2 (1.0-4.2) kPa) (P < 0.001), and when the tip was situated at the mid-tracheal level there was a further increase to 24 (12-50) mmHg (2.4 (1.2-4.9) kPa) (P < 0.001). Maximal gas leakages were median 1 (0-2) l/min-1. PEEP at the mid-tracheal level was 3 (0-7) cmH2O (0.3(0-0.7) kPa). When 15 min of the procedure had elapsed, PaO2 was 232 (112-350) mmHg (30.9 (14.9-46.6) kPa) and PaCO2 39 (33-46) mmHg (5.2(4.4-6.1) kPa). The lowest oxygen saturation was median 98 (96-100)% and the highest end tidal CO2 34 (24-41) mmHg (4.5(3.2-5.5) kPa). It was easy to examine the laryngeal opening and a good assessment of vocal cord function was allowed when muscle relaxation ceased. We conclude that flexible fibreoptic bronchoscopy through the laryngeal mask is a safe technique provided that total intravenous anaesthesia is used. It is a valuable alternative to flexible bronchoscopy performed with topical anaesthesia. PMID- 8424287 TI - Postoperative decline in plasma aspirin-esterase and cholinesterase activity in surgical patients. AB - We studied preoperative and postoperative plasma hydrolysis of aspirin and plasma cholinesterase activity in surgical patients. Postoperative aspirin esterase and cholinesterase activities fell sharply (119 +/- 32 micrograms ml h-1 and 3746 +/- 1068 U l-1 respectively; P < 0.01) in comparison to basal preoperative values (157 +/- 33 micrograms ml h-1 and 4620 +/- 940 U l-1 respectively, P < 0.01). We suggest that alterations in plasma esterase during the postoperative period may be related to the metabolic response to anesthesia- and surgery-induced stress. PMID- 8424288 TI - Preoxygenation techniques: the value of nitrous oxide. AB - Changes in arterial oxygen saturation during induction of anaesthesia and intubation were studied using the pulse oximeter. Seventy-five young ASA I patients undergoing elective uncomplicated surgery were divided equally into three groups. The patients were preoxygenated with 100% oxygen, 50% oxygen: 50% nitrous oxide or 30% oxygen: 70% nitrous oxide for 1 min. All were then induced with thiopentone, paralysed with suxamethonium and orally intubated. Arterial oxygen saturations were continuously recorded by a separate investigator. All groups showed similar arterial desaturation during suxamethonium-induced apnoea and intubation, but the degree of desaturation was not clinically significant and no patient showed clinical signs of hypoxaemia. Preoxygenation with mixtures of oxygen and nitrous oxide can hasten the build-up of alveolar nitrous oxide concentration and help to smooth induction without compromising oxygenation of patients. PMID- 8424289 TI - Verification of the position of a central venous catheter by intra-atrial ECG. When does this method fail? AB - The purpose of this study was to determine why intra-atrial ECG tracing for checking the position of a central venous catheter fails in certain patients. Three hundred and fifty prospective and consecutive patients scheduled for central venous catheterization using various puncture sites and techniques were investigated. The catheters were 20 cm in length. After its introduction, the catheter was connected to an Alphacard (Sterimed, Saarbrucken) for the intra atrial ECG tracing. The method failed in 29 patients, of whom nine had manifest myocardial pathology. In two patients the catheter looped, while in the remaining 18 the catheter proved to be too short. In these 18 patients, the cannulation was mainly performed via the external jugular vein and/or from the left side. Most of the patients were elderly males, and 11 of the 18 patients showed radiological signs of pulmonary emphysema. In such individuals it is advisable to use a catheter longer than 20 cm. PMID- 8424290 TI - Psychomimetic reactions after neurolept and propofol anaesthesia. AB - The purpose of this study was to compare the frequency of psychomimetic reactions after 24 h and 3 months following total intravenous anaesthesia with propofol and neurolept anaesthesia. Forty otherwise healthy female patients were randomly divided into two groups. All were undergoing elective gynaecological laparotomy for non-malignant disease. Nineteen patients were anaesthetized with droperidol, fentanyl, pancuronium, N2O/O2. Twenty patients received total intravenous anaesthesia with propofol, fentanyl and pancuronium. Twenty-four hours after the anaesthesia the patients were interviewed about their subjective experiences of anaesthesia and recovery. Three months after the operation the patients were sent a questionnaire concerning ability to work, sleep and memory disorders. After 24 h the anaesthesia was judged as good by 18 patients receiving propofol and 13 patients receiving NLA (n.s.). The recovery was judged as good by 16 patients in the propofol group and six patients in the NLA group (P < 0.05). Locked-in feelings were reported by one patient in the propofol group and ten patients in the NLA group (P < 0.01). Impairment of memory was reported by one patient in the propofol group and seven patients in the NLA group (P < 0.01). A questionnaire used after 3 months was answered by 18 patients in the propofol group and 17 patients in the NLA group. There were few complaints, and no differences were found between the two groups. In conclusion, total intravenous anaesthesia with propofol seems more acceptable than anaesthesia with neurolept as judged by the patients 24 h after anaesthesia. There were no differences between the two groups concerning psychomimetic reactions 3 months after anaesthesia. PMID- 8424291 TI - Recall of music: a comparison between anaesthesia with propofol and isoflurane. AB - Sixty patients undergoing laparoscopy were randomly assigned to receive total intravenous anaesthesia with propofol or inhalation anaesthesia with isoflurane. Patients in these two groups were also randomly assigned to three subgroups listening to soft music, hard rock music or no music at all. Twenty-four hours after surgery all patients were interviewed and asked if they had heard music during the operation. A tape with seven different pieces of music was also played for the patients. The music they heard during surgery was one of these. No patient had any memories or experiences from the operation. Four patients had dreams, three from the total intravenous anaesthesia group and one from the inhalation anaesthesia group. Twelve patients believed they had heard music, ten from the total intravenous anaesthesia group and two from the inhalation anaesthesia group (P < 0.05). Two patients, one from each group, picked the right melody. In conclusion, there was no difference between the two groups with respect to recall during anaesthesia. Patients in the TIVA group were significantly more inclined to state that they had been exposed to music. PMID- 8424292 TI - Influence of high thoracic epidural anesthesia on left ventricular contractility assessed using the end-systolic pressure-length relationship. AB - The effect of high thoracic epidural anesthesia (TEA on left ventricular contractility was studied in a prospective clinical trial. Forty-eight patients with ASA physical status 1 and 2 and without cardiovascular disease were included in the study. Thirty-six patients scheduled for elective upper abdominal surgery were randomly assigned to Group 1 (TEA, bupivacaine 0.25%, n = 12), Group 2 (TEA, bupivacaine 0.5%, n = 12) or to Group 3 (control without TEA, n = 12). TEA induced a sensory block which extended over all cardiac segments. In order to assess the effect of systemically absorbed bupivacaine, we studied a separate group of patients who received lumbar epidural anesthesia without involvement of the cardiac segments: Group 4 (LEA, bupivacaine 0.5%, n = 10). Left ventricular contractility was assessed using the end-systolic pressure-length relationship. Left ventricular dimensions were measured by transesophageal echocardiography. All hemodynamic measurements were performed under general anesthesia. There was no significant difference in systolic or diastolic arterial pressure, heart rate, left ventricular end-systolic and end-diastolic cross-sectional areas and left ventricular wall stress between the four groups. Left ventricular maximum elastance as a measure of left ventricular contractility was significantly (P < 0.001) reduced in Groups 1 and 2 [8.1 (+/- 3.5) and 9.6 (+/- 4.4) kPa.cm-1, respectively] as compared to Groups 3 and 4 [18.4 (+/- 8.8) and 17.7 (+/- 7.7) kPa.cm-1, respectively]. No significant difference could be demonstrated between Groups 1 and 2 or between Groups 3 and 4. It is concluded that high TEA severely alters left ventricular contractility even in subjects without pre-existing cardiac disease. PMID- 8424293 TI - The effects of prophylactic dixyrazine on postoperative vomiting after two different anaesthetic methods for squint surgery in children. AB - The incidence of postoperative vomiting after squint surgery was studied for two anaesthetic techniques with and without prophylactic dixyrazine. After induction, anaesthesia was maintained with either fentanyl/pancuronium/nitrous oxide or halothane/nitrous oxide in two randomly selected groups of 58 children each. Half of the children in each group were randomly allocated to receive dixyrazine 0.25 mg kg-1 i.v. after surgery had been completed but before reversal of muscle relaxants or termination of anaesthesia. With prophylactic dixyrazine the incidence of postoperative vomiting was significantly reduced from 69% (20/29) to 21% (6/29) in the fentanyl group and from 45% (13/29) to 10% (3/29) in the halothane group. Without prophylactic dixyrazine, 20 of 29 children in the fentanyl group vomited compared to 13 of 29 in the halothane group (n.s.). Thus, prophylactic dixyrazine reduced the incidence of vomiting in children given either opioid or halothane anaesthesia for squint surgery. In comparable groups avoidance of opioid anaesthetic technique and use of prophylactic dixyrazine resulted in a greatly reduced incidence of vomiting. PMID- 8424294 TI - Analgesic action of metoclopramide in prosthetic hip surgery. AB - Prosthetic hip surgery was performed under subarachnoidal anaesthesia with bupivacaine 16-20 mg and morphine 0.2 mg. Preoperatively, metoclopramide 1 mg.kg 1 was given i.v., followed by an infusion of 1.5 mg.kg-1 over 9 h (n = 17). Control patients received corresponding volumes of solvent (n = 23. The design of the study was double blind. The characteristics of the spinal block (level of analgesia to pinprick and muscular block) and postoperative VAS pain scores were similar in both groups. During the 24 h following the start of the infusion, four patients receiving metoclopramide required i.v. opioids, compared to 15 in the control group (P < 0.05). The pain-free period was longer (P < 0.05) in the metoclopramide group. Arterial PCO2-levels were increased, reaching a maximum within 6 h of infusion, with no significant difference between the groups. The study suggests an analgesic action of metoclopramide. PMID- 8424295 TI - Effects of intrathecal morphine and spinal anaesthesia on sympathetic nerve activity in humans. AB - Microneurography with direct registration of efferent sympathetic nerve activity to muscle or skin was used to study the effect of morphine and procaine given intrathecally to patients scheduled for transurethral resection of the prostate. A lumbar spinal catheter was inserted and multiunit postganglionic sympathetic activity was recorded in a muscle or skin fascicle of the peroneal nerve. After injection of 0.4 mg morphine, sympathetic nerve activity to muscle (n = 6) or skin (n = 2) was recorded for 30 min. Compared with a 10-min control period prior to the injection no change in activity could be observed. In contrast, spinal anaesthesia (procaine 75-100 mg) with an upper level of sensory blockade at T3 T11 completely eliminated intraneurally recorded sympathetic activity within 5 min (n = 8). Skin vasodilation and loss of sudomotor responses in the foot accompanied the neural blockade. PMID- 8424296 TI - Renal vascular response to left atrial injection. An experimental study in the pig. AB - It has previously been shown that a rapid, transient and pronounced reduction in superficial renal cortical blood flow can be elicited by injection of 1 ml of plasma into the left atrium of the anaesthetized pig. This renal vascular response might constitute an important error in measurements of renal blood flow by means of the microsphere technique. In the present study we have investigated total renal (ultrasonic Doppler) and superficial renal cortical (laser Doppler flowmetry) blood flow after left atrial injections of saline and autologous blood. The average reductions in total renal blood flow were 4.7 +/- 2.4% (mean +/- s.d.) and 18.4 +/- 12.5% (mean +/- s.d.) for the blood and saline injections, respectively. The flow reduction was more pronounced in the superficial renal cortex with average reductions of 17.4 +/- 26.0% (mean +/- s.d.) and 59.4 +/- 38.5% (mean +/- s.d.) for blood and saline, respectively (laser Doppler flowmetry). Furthermore, two consecutive injections of microspheres were performed. The results from the second injection were compared with those from the first injection. In the superficial renal cortex the second injection indicated a significant reduction in blood flow and an significantly increased heterogeneity in the blood flow in two different tissue volumes. No corresponding differences were found in the juxtamedullary cortex. These results indicate that the renal vascular response to a left atrial injection is detectable as a reduction in total renal blood flow and is most pronounced in the superficial renal cortex. All these results taken together constitute an important error in measurements of superficial renal cortical blood flow by the microsphere technique. PMID- 8424297 TI - Blood pressure and heart rate during orthostatic stress and walking with continuous postoperative thoracic epidural bupivacaine/morphine. AB - Thirty-one patients scheduled for elective cholecystectomy performed through a mini-laparotomy, were randomized to received either combined thoracic epidural anaesthesia/light general anaesthesia and postoperative balanced analgesia with continuous epidural bupivacaine 10 mg.h-1 and morphine 0.2 mg.h-1 for 38 h after surgery plus systemic ibuprofen 600 mg x 8 h-1 (N = 15) or general anaesthesia and postoperative analgesia with systemic morphine and ibuprofen 600 mg x 8 h-1 (N = 16). During postoperative epidural infusion sensory blockade to pinprick was Th4 to L1, and analgesia at rest and during mobilisation was superior compared to systemic morphine and NSAID. There were no significant differences between groups in haemodynamic responses (BP and heart rate) during rest, orthostatic stress and after walking assessed before, 24 and 48 h after operation except for a clinically unimportant lower heart rate (approximately 10 bpm) 48 h after surgery at rest and during orthostatic stress in the epidural group. There was no significant difference between groups in number of patients with a reduction > 20 mmHg (2.7 kPa) in systolic blood pressure during orthostatic stress (two in each group at 24 h) or in number of episodes of dizziness, nausea or vomiting during rest or mobilisation. These results do not support the common belief that low dose thoracic epidural bupivacaine/morphine may prevent ambulation due to sympathetic blockade or to impaired cardiovascular adaptation to the upright position. PMID- 8424298 TI - Effects of deliberate hypotension induced by labetalol with isoflurane on neuropsychological function. AB - The effect of deliberate hypotension on brain function measured by neuropsychological tests was studied in 41 adult patients. Twenty-four patients were anaesthetized for middle-ear surgery with deliberate hypotension induced by labetalol with isoflurane (hypotensive group). Seventeen patients without hypotension served as a control group. The mean arterial pressure was 77 +/- 2 mmHg (10.3 +/- 0.3 kPa) before hypotension and 50 +/- 0 mmHg (6.7 +/- 0.0 kPa) during hypotension in the hypotensive group, and 86 +/- 2 mmHg (11.5 +/- 0.3 kPa) during anaesthesia in the control group. The following psychological tests were performed: four subtests of the Wechsler Adult Intelligence Scale (similarities, digit span, vocabulary and digit symbol), Trail-Making tests A and B, Zung tests (self-rating anxiety scale and self-rating depression scale) and two-part memory test battery with immediate and delayed recall. The tests were performed preoperatively and 2 days postoperatively. There were no statistically significant differences between the groups in any of the tests in the changes from preoperative value to postoperative value. The results indicate that hypotension induced by labetalol with isoflurane has no significant harmful effects on mental functions compared to normotensive anaesthesia. PMID- 8424299 TI - Duration of experimental nerve block by combinations of local anesthetic agents. AB - The effects of bupivacaine-prilocaine and meperidine-lidocaine combinations (as compared with those of the agents used alone) on the duration of peripheral sensory nerve block were studied with the infraorbital nerve block model (IONB) in the rat, and those on motor block with spinal anesthesia (SA) in the mouse. The duration of bupivacaine-induced IONB was invariably prolonged when prilocaine was included in the solution. When included in 0.125% bupivacaine, 1.0% prilocaine had a slightly less pronounced enhancing effect than 0.5% prilocaine (24-57% vs. 74%-104%, respectively). The duration of IONB with 1.0% prilocaine was significantly reduced (14-37%) by inclusion of 0.125% bupivacaine. In SA, inclusion in 0.125% bupivacaine of prilocaine (0.5% or 1.0%) prolonged motor block by 128% and 192%, respectively. When included in 0.25% bupivacaine, both 0.5% and 1.0% prilocaine significantly reduced the duration of SA, by 42% and 37%, respectively. With one exception, the duration of IONB by meperidine was significantly shortened (< 44%) when lidocaine was included in the solution. In SA, inclusion of 2% lidocaine with 2% meperidine did not affect the duration of meperidine-induced motor block. The duration of SA obtained with the combination of 4% lidocaine and 4% meperidine was 45% shorter than that induced by 4% meperidine alone. The reasons for these variable effects are not clear, but may be due to interaction or antagonism at any of multiple sites. PMID- 8424300 TI - The cardiovascular response to ketamine: the effects of clonidine and lignocaine. AB - The effectiveness of clonidine or lignocaine in reducing the cardiostimulatory effects of ketamine was studied. A double-blind, randomized design was used in three groups of 20 patients each, of ASA grade 1 or 2 presenting for minor elective surgery. The clonidine group received 0.3 mg clonidine orally with the premedication while the lignocaine group received 1.5 mg.kg-1 lignocaine prior to induction. The third group formed the controls. All patients were induced with 2 mg.kg-1 ketamine intravenously. The systolic blood pressure in the clonidine group was significantly lower than that of both the control and lignocaine groups throughout the study (P < 0.05). It was concluded that clonidine was effective in reducing the hypertensive response to ketamine, whereas lignocaine had no effect. PMID- 8424301 TI - Coronary artery spasm with ventricular tachycardia after administration of methoxamine during cervical epidural anesthesia--a case report. AB - Coronary artery spasm occurred during thoracotomy under cervical epidural anesthesia in a 60-year-old male patient who had no prior history of myocardial ischemia. It is most likely that the administration of methoxamine induced the spasm. Hypotension and venodilatation induced by the epidural anesthesia and increased vagal tone might also contribute to the spasm. PMID- 8424302 TI - The calcium channel blocker nifedipine fails to inhibit leucocyte elastase release during cardiopulmonary bypass. AB - Circulating concentrations of leucocyte elastase were measured in 16 adult patients undergoing cardiopulmonary bypass (CPB) with a flat-sheet membrane oxygenator. Eight patients (Group I) received the calcium channel blocker nifedipine (9 micrograms.kg-1 x h-1) during CPB. Eight patients (Group II) did not receive any calcium channel blocker during surgery and served as the control group. Elastase concentrations were measured at 7 time points: 2 before, 2 during, and 3 after CPB. The bypass procedure was associated with elevation in elastase concentrations (P < 0.001). Comparing to baseline values elastase concentrations were significantly elevated (P < 0.05) 60 min after the start of CPB and on all measurements done after CPB. Elastase concentrations correlated with the duration of CPB (rs = 0.76, P < 0.001), and were not influenced by nifedipine infusion as revealed by comparing the two groups. This study demonstrates moderate elastase release during CPB with a flat-sheet membrane oxygenator and fails to confirm inhibition of elastase release by nifedipine infusion during CPB. PMID- 8424303 TI - Cerebral haemodynamic and electrocortical CO2 reactivity in pigs anaesthetized with fentanyl, nitrous oxide and pancuronium. AB - Cerebral haemodynamic, metabolic and electrocortical reactivity to alterations in arterial CO2 tension (PaCO2) was assessed in seven mechanically ventilated juvenile pigs to test an experimental model designed for cerebral pharmacodynamic and pharmacokinetic studies. The animals were anaesthetized with fentanyl, nitrous oxide and pancuronium and sequentially normo- and hyperventilated over a 100-min period. Five measurements were made at 25-min intervals. The cerebral blood flow (CBF) was measured with the intra-arterial 133Xe technique and the cerebral metabolic rate for oxygen (CMRO2) determined from CBF and the cerebral arteriovenous oxygen content difference. A linear correlation (r = 0.845) was found between CBF and PaCO2. The cerebrovascular reactivity to hypocapnia (delta CBF/delta PaCO2) was maintained throughout the experimental period and amounted to (95% confidence interval) 9.1 (7.1-11.1) ml x 100 g-1 x min-1 x kPa-1 within the PaCO2 range 3.3-6.3 kPa. The CMRO2 was not influenced by hyperventilation. The baseline electroencephalographic (EEG) pattern was stable at normocapnia (mean PaCO2 5.6 kPa), whereas spectral values for delta and total average voltage increased significantly (P < 0.05) at extensive hypocapnia (3.5 kPa). Maintenance of cerebral CO2 reactivity and spectral EEG voltage at a stable plasma level of fentanyl is complementary to the cerebral haemodynamic and metabolic stability previously found at sustained normocapnia in this model. PMID- 8424304 TI - Myocardial ischaemia as judged from transoesophageal echocardiography and ECG in the early phase after coronary artery bypass surgery. AB - The incidence of myocardial ischaemia, as diagnosed by transoesophageal echocardiography (TEE) versus ECG, and the relationship between ischaemic events and haemodynamic parameters were studied in 30 patients in the early phase after coronary artery bypass grafting. Information comprising invasive haemodynamics, TEE measurements, and 12-lead ECG was obtained on arrival of the patient in the intensive care unit (ICU), and then hourly in the ICU for 5 h. In the ICU, TEE signs of ischaemia were found in 14 patients and ECG signs of ischaemia in six patients. The ischaemic events were not related to levels of blood pressure or heart rate. Three patients showed signs of myocardial infarction postoperatively. All three of these patients showed both TEE and ECG signs of ischaemia in the ICU. It was concluded that TEE reveals more ischaemic events than ECG in the early postoperative period and that these ischaemic events do not correlate with the haemodynamic indices. PMID- 8424305 TI - Intra-arterial papaverine and leg vascular resistance during in situ bypass surgery with high or low epidural anaesthesia. AB - In situ saphenous vein arterial bypass flow was studied in 16 patients with respect to level of epidural anaesthesia. Arterial pressure and electromagnetic flow were used to evaluate arterial tone by intra-arterial (i.a.) papaverine. Eight patients had a low epidural block (< or = Th. 10) and eight patients were operated during high epidural anaesthesia (> Th. 10). Flow increased and arterial pressure decreased after i.a. papaverine in all patients. When compared with patients operated during high epidural anaesthesia, flow increase and decrease in vascular resistance took place in patients operated during low epidural anaesthesia (P < 0.02). Increase in arterial flow after i.a. papaverine was not significantly different in patients operated in low epidural and general anaesthesia (n = 8). In eight patients with insulin-dependent diabetes mellitus who had low epidural anaesthesia, the increase in flow after i.a. papaverine was not significantly different to that noted during high epidural anaesthesia. The results indicate that the level of analgesia influences graft flow after i.a. papaverine, probably reflecting differences in the effect of epidural anaesthesia on sympathetic tone to the leg. PMID- 8424306 TI - Verbal learning differences in epileptic patients with left and right temporal lobe foci--a pharmacologically induced phenomenon? AB - A total of 27 patients with medically intractable complex partial seizures has been investigated for effects of anticonvulsant drugs on mental abilities, particularly on verbal memory performance. Fourteen patients with right (RTLE) and 13 with left (LTLE) temporal lobe epilepsy have been tested with a word list learning paradigm under the conditions of full and reduced anticonvulsant medication. Memory performance has significantly improved with drug reduction, however only for the LTLE group. In addition, significant group differences for verbal memory between LTLE and RTLE subjects under full medication have completely disappeared with drug reduction. Finally this investigation demonstrates, that very specific and circumscribed steps of verbal memory processing, particularly retrieval abilities after interference, are affected by anticonvulsants. These findings underline the importance of pharmacological effects on cognition and suggest to reevaluate their relevance compared to other contributing factors. PMID- 8424307 TI - "Panic attacks" in Parkinson's disease. A long-term complication of levodopa therapy. AB - A series of 31 Parkinson's disease (PD) patients suffering from panic attacks (PA), late in the evolution of their disease, was analyzed from a group of 131 levodopa-treated PD patients. We found that many of motor, sensory, and vegetative symptoms, previously described as complicating phenomena in PD, constituted some of the symptoms of panic disorders. Comparing PA series with the series of PD patients who did not complain of PA, we discovered a clear-cut relationship of PA with the presence of standing/gait troubles (p < 0.001), depression (p < 0.001), and dyskinesias/fluctuations (p < 0.001). The patients of the PA series also presented a more precocious age of PD onset, were put on levodopa therapy earlier, and needed to be treated with higher doses of levodopa than the patients without PA. Finally, we hypothesize that PA could be considered to be a sort of abstinence syndrome from levodopa, because they appears mostly (90.3%) in the OFF phase of fluctuations, and are relieved administering new doses of levodopa or dopaminergic agonists. Nevertheless, we suggest PA are not directly related to the pharmacological properties of levodopa, but to alterations of the noradrenergic systems in the CNS. PMID- 8424308 TI - CSF cholinesterase in early-onset and late-onset Alzheimer's disease and multi infarct dementia of Chinese patients. AB - Using Ellman spectrophotometric method we measured the total cholinesterase (ChE) activity in lumbar cerebrospinal fluid (CSF) of 13 persons without neurological disorder, 10 non-demented patients with cerebral infarcts, 17 patients with dementia of Alzheimer's type (DAT) (11 presenile, 6 senile cases), 10 patients with multi-infarct dementia (MID), 1 patient with Parkinson's disease associated with dementia. The ChE activity in CSF was significantly lower in the DAT group compared with age-matched control subjects (p < 0.001). This paper also analyses the possibility of using CSF ChE activity as a marker of DAT, and the relationships between its level of activity and the age of the patient at onset, stage of illness and severity of dementia as well as discrepancies in the data published so far. Previous work has shown that ChE activity in the brain tissue and CSF of MID is normal: therefore, if low ChE activity is found in the CSF of MID patients, as was obtained in 8 out of 10 cases in our series, the diagnosis of mixed dementia should be considered. PMID- 8424309 TI - Paramyotonia congenita (Eulenburg): clinical, neurophysiological and muscle biopsy observations in a Swedish family. AB - A Swedish family with Paramyotonia congenita (Eulenburg) (PMC) is presented. Clinical neurological examination, neurophysiological examination (n = 5) and muscle biopsy (n = 4) were performed. Different clinical features were found in various combinations in the individual family members. The clinical symptoms were: (1) cold-induced myotonia, (2) attacks of weakness, (3) persistent weakness and (4) no symptoms but other signs of muscle affection. In the patients with myotonia, the neurophysiological examination showed spontaneous myotonic discharges which were frequent at room temperature but disappeared after cooling. Furthermore, the amplitude of M. abductor digiti minimi compound action potential, during supramaximal ulnar nerve stimulation, decreased significantly after cooling. In the patients with persistent weakness there were no spontaneous myotonic discharges, but myopathic abnormalities were found in proximal muscle. In the patients with myotonia as well as in the patients with manifest muscle weakness, muscle biopsy showed a variation of muscle fibre diameters, centrally located nuclei, occasional atrophic fibers and an atrophy of type IIB muscle fibres. These findings are unspecific but have been described in PMC patients in earlier studies. An ancestor to the family, who had myotonia, lived in the same town and at the same time as Albert Eulenburg, which may suggest that this family is a part of the originally described family (1). PMID- 8424310 TI - Accuracy of self-assessment of the minimal record of disability in patients with multiple sclerosis. AB - We tested the validity of a self-administered version of the minimal record of disability (MRD) for multiple sclerosis (MS) by measuring the agreement level between patients' self-assessment and neurologists' independent ratings. 96 MS patients and 4 neurologists took part in the experiment; the agreement level was measured in terms of the intraclass correlation coefficient (ICC). On the Kurtzke functional system (FS) the ICC ranged from 0.26 in the sensory to 0.69 in the pyramidal function; a high concordance (ICC = 0.84) was found on the expanded disability status scale (EDSS). The ICC values were above 0.70 for most of the incapacity status scale (ISS) and environmental status scale (ESS) items. A modified, self-administered version of the MRD may represent a reliable instrument for obtaining a comprehensive profile of patients' abilities. PMID- 8424311 TI - Electrophysiological study of neurologically asymptomatic HIV1 seropositive patients. AB - EEGs, brainstem auditory evoked potentials (BAEPs) and auditory event-related potentials (ERPs) were recorded from 33 individuals infected with the human immunodeficiency virus, type 1 (HIV1+ patients: 13 CDC Class II or III; 20 Class IV). All were neurologically asymptomatic, non-demented, and had a past history of intravenous drug abuse. Sixteen age- and sex-matched normals and 10 HIV1- former drug addicts served as controls. Half of the HIV1+ and HIV1- subjects displayed mild EEG anomalies and, except for one HIV1+ patient, BAEPs were normal in both groups. ERPs were normal in all HIV1- subjects but anomalous (longer latencies of components P2, N2, P3; reduced amplitude of P3) in 9 HIV1+ patients (27%), the incidence of such anomalies being higher for Class IV than Class II/III patients. Auditory ERPs proved the most sensitive and specific of these electrophysiological procedures in detecting subclinical central nervous system involvement in HIV1 infection. PMID- 8424312 TI - Memory impairment in Cushing's disease. AB - In the present study the cognitive performance of 25 patients with Cushing's disease (CD) was extensively evaluated in comparison with normal control subjects, matched one by one. The results indicate a selective impairment of memory functions: the number of patients showing a significantly impaired mnesic performance increases with age. Moreover, the neuropsychological impairment tends to recover in those cases who underwent further controls after surgical treatment. The neuropsychological data are discussed in the light of recent evidence in the literature concerning the effects of adrenal steroids on the brain. PMID- 8424313 TI - X-linked bulbospinal muscular atrophy (Kennedy's syndrome): a report of three cases. AB - Two cases of X-linked bulbo-spinal muscular atrophy and one sporadic case with the same clinical features are presented. All three cases were extensively studied by electrophysiological methods. One of the patients showed some transient weakness, which was partly improved by pyridostigmin. In this patient the Decrement test and jitter revealed a mild degree of motor-end plate dysfunction. Clinical and electrophysiological findings obtained from all three patients suggest that in Kennedy syndrome cell bodies of group IA muscle afferents are also affected selectively, with other peripheral afferents. PMID- 8424314 TI - Antioxidant enzymatic activities after experimental subarachnoid hemorrhage in rats. AB - Lipid peroxidation has been hypotesized as one of possible factors involved in the pathogenesis of neuronal damage and delayed vasospasm after subarachnoid hemorrhage. In the brain there are anti-oxidant enzymatic systems which act as scavengers of superoxides and free radicals. In the present study the pattern of enzymatic anti-oxidant activities (Cu-Zn and Mn superoxide dismutase, and glutathione peroxidase) was investigated in an experimental model of subarachnoid hemorrhage in the rat in order to verify whether the hemorrhagic insult may be responsible for an impairment of such anti-oxidant systems. Enzymatic activities were assayed in three different rat brain areas (cerebral cortex, hippocampus and brain stem) of sham-operated and at 30 min, 1, 6 and 48 h after subarachnoid hemorrhage induction. After the hemorrhage induction the Cu-Zn superoxide dismutase activity in cerebral cortex was significantly reduced at all the set times (p < .05), while Mn-superoxide dismutase activity was significantly decreased since 1 h (p < .05) until 48 h (p < .05). Glutathione peroxidase activity was significantly reduced only in the late phase (48 h) of subarachnoid hemorrhage (p < .01). In the hippocampus, all enzymatic activities were significantly reduced in the late phase. In the brain stem Cu-Zn superoxide dismutase was significantly impaired at 1 and 6 h (p < .05) after subarachnoid hemorrhage induction, while in the late phase (48 h) reached the control value. The mitochondrial Mn-superoxide dismutase was significantly reduced since 1 h (p < .05) until 48 h (p < .02) after subarachnoid hemorrhage.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424315 TI - Hepatitis A post-viral encephalitis. AB - We report a seven-year-old girl who developed a hepatitis A viral infection and encephalitis. The patient developed fever, abdominal pains and jaundice. Five days later she became delirious, combative, and did not respond to verbal commands. Laboratory studies showed elevated liver enzymes and elevated serum immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies to hepatitis A virus. Cerebrospinal fluid contained IgG antibodies to hepatitis A virus but not IgM antibodies. Polymerase chain reaction, which amplifies a portion of the hepatitis A virus genome, did not demonstrate viral nucleic acid in cerebrospinal fluid. These studies suggest that the patient may have suffered from a post-viral hepatitis A encephalitis from which she fully recovered. PMID- 8424316 TI - Multifocal inflammatory leukencephalopathy caused by adjuvant therapy with 5 fluorouracil and levamisole after resection for an adenocarcinoma of the colon. PMID- 8424317 TI - Mortality rates for parkinsonism in Italy (1969 to 1987). AB - Parkinsonism death rates in Italy were analysed for the period 1969-1987. Crude death rates increased markedly in the above period for both sexes. Age-specific rates underwent important changes during these twenty years. In the years 1969-71 the age-specific death rates reach their maximum values at ages 80-84, after which they decline. In the years 1983-87 the rates continue to increase until the 80-84 age group, when they reach a plateau. Moreover, in the more recent years the age-specific rates for ages < 70-75 are lower, whereas those for the older ages are approximately twice as high as the values in the earlier years. The age adjusted death rates (1981 Italian population) increased slightly in the period studied for both sexes. Rates for males exceed those for females in all the previous analyses. Minor differences appeared in death distribution between the five large areas Italy was divided into. The observed changes in death rates are likely to reflect: changes in age composition of the population over time, greater attention to parkinsonian patients and especially increased duration of this disease following the discovery of its neurochemical basis and effective medical treatment. PMID- 8424318 TI - Anxiety, impulsivity and depressed mood in relation to suicidal and violent behavior. AB - Many different variables have been associated with suicidal behavior as well as with violent behavior. This study was designed to test a model that attempts to relate such variables to violence and suicide risk. Sixty psychiatric patients were evaluated and divided into suicidal and nonsuicidal groups and then into violent and nonviolent groups. Scores on suicide risk, violence risk, anxiety, anger, impulsivity and mood were then compared between these groups. In a second analysis, scores on suicide risk and violence risk were correlated with all the other variables. Anxiety and impulsivity were found to strongly correlate with suicide risk. Angry and resentful mood correlated with violence risk; trait anxiety correlated negatively with violence risk. The results contribute further insight into the authors' two-stage model of countervailing forces. PMID- 8424319 TI - Cognitive impairment, dementia and psychosocial functioning in human immunodeficiency virus infection. A prospective study based on DSM-III-R and ICD 10. AB - Progressive cognitive impairment in human immunodeficiency virus (HIV) infection, called acquired immunodeficiency syndrome (AIDS) dementia complex (ADC), significantly influences the social prognosis of afflicted patients. The frequency and character in different stages of the infection are controversially discussed. In previous studies, differences in the selection of patients and methods of testing led to widely differing results. For these reasons, in the present prospective study on 45 HIV-infected patients, a structured psychiatric interview (SIDAM) was conducted based on the algorithm of diagnosing dementia in DSM-III-R and the ICD-10 guidelines. The psychopathological findings are expressed in syndrome scores; the results are summarized in a total score (SISCO). The interview contains the Mini-Mental State Examination. The degree of psychosocial functioning was estimated on the global assessment of functioning, Axis V of DSM-III-R. In stages preceding AIDS, only slight cognitive dysfunction was found compared with age- and education-matched normal controls, and this caused no relevant disturbance of psychosocial functioning. In 9 patients with manifest AIDS, dementia was diagnosed with DSM-III-R criteria and ICD-10 guidelines (30% of the AIDS patients). They showed marked impairment of intellectual ability, memory, verbal ability and calculation and constructional ability and fewer cortical focal symptoms (aphasia and apraxia). Corresponding to previous studies, major cognitive dysfunction in HIV infection can be characterized as subcortical dementia. PMID- 8424320 TI - Two subtypes of adolescent suicide attempters. An empirical classification. AB - An empirical classification of 203 adolescent suicide attempters (mean age 17.3) was achieved using multiple correspondence analysis. The characteristics upon which the classification is based concern sociodemographic as well as psychological variables. Two groups are identified: the first is predominantly characterized by recent problematic behaviors, whereas the second group is primarily characterized by problematic circumstances. Analyzing related variables, the first group seems to have special clinical and preventive interest, because of its high risk for recidivism. The second group seems to have a satisfactory level of functioning. PMID- 8424321 TI - Relationship between personality and debrisoquine hydroxylation capacity. Suggestion of an endogenous neuroactive substrate or product of the cytochrome P4502D6. AB - We administered the Karolinska Scales of Personality to 225 healthy subjects in Spain selected from a group of 925 individuals previously phenotyped with regard to their capacity to hydroxylate debrisoquine. A significant relationship was found between the scores in as many as 4 of the 15 subscales (psychic anxiety, psychasthenia, inhibition of aggression and socialization) and the debrisoquine hydroxylation capacity. Poor metabolizers were more anxiety-prone and less successfully socialized than extensive metabolizers of debrisoquine. This and a previous study among subjects in Sweden suggest that there may be a relationship between personality and the activity of the enzyme hydroxylating debrisoquine (cytochrome P4502D6). This polymorphic enzyme may have an endogenous neuroactive substrate or product, such as a biogenic neurotransmitter amine. PMID- 8424322 TI - A case-control study of family history and cerebral cortical abnormalities in schizophrenia. AB - The relationship between family history of psychosis and cortical sulcal widening on CT scans was investigated. Forty-two schizophrenic subjects with a positive family history were individually matched to 42 schizophrenics with no documented family history and 42 healthy controls. Abnormally wide sulci, as defined on the basis of the normal control data, were significantly more common in the family history negative group (9/42) than in the family history positive group (1/42) or the controls (2/36). No significant correlation was found between cerebral cortical ratings and ventricular size. PMID- 8424323 TI - Bereavement among elderly people: grief reactions, post-bereavement hallucinations and quality of life. AB - Ratings of grief reactions, post-bereavement hallucinations and illusions and quality of life were made during the first year after the death of a spouse among 14 men and 36 women in their early seventies. In both sexes, the reactions were generally moderate or mild and characterized by loneliness, low mood, fatigue, anxiety and cognitive dysfunctioning. Feeling lonely was the most persistent problem during the year. Post-bereavement hallucinations or illusions were very frequent and considered helpful. Half of the subjects felt the presence of the deceased (illusions); about one third reported seeing, hearing and talking to the deceased (hallucinations). Former marital harmony was found to make a person more prone to loneliness, crying and hallucinations or illusions. The quality of life was significantly lower among the bereaved than among married people and those who never married, but equalled that found among divorcees. PMID- 8424324 TI - Thoracic metastases from carcinoma of the nasopharynx: high frequency of hilar and mediastinal lymphadenopathy. AB - OBJECTIVE: Nasopharyngeal carcinoma is a malignant tumor commonly encountered in Chinese patients living in or originating from Hong Kong or southern China. This article describes the previously unreported radiologic appearances of thoracic metastases from nasopharyngeal carcinoma. MATERIALS AND METHODS: The radiographic (33 patients) and CT (eight patients) appearances of thoracic metastases from nasopharyngeal carcinoma were studied retrospectively. All 33 patients had biopsy proved primary nasopharyngeal carcinoma, and seven patients had biopsy-proved thoracic metastases. Radiologic and clinical evidence of metastases was unequivocal in 26 others, 16 of whom had synchronous spread to cervical lymph nodes, bone, or liver. RESULTS: Twenty-one patients (64%) had evidence of thoracic lymphadenopathy, most frequently hilar (n = 18, 55%), on chest radiographs or CT scans. Seventeen patients (52%) had evidence of multiple parenchymal pulmonary metastases. Enlargement of lymph nodes without multiple pulmonary deposits was seen in 12 patients (36%), seven of whom had radiologic signs of bronchial obstruction, hemoptysis, or a single pulmonary lesion simulating a synchronous bronchial neoplasm. Pleural effusions or deposits (n = 6), lymphangitis carcinomatosa (n = 5), and rib metastases (n = 4) also were seen. CONCLUSION: Metastases from nasopharyngeal carcinoma may be detected in a wide range of thoracic sites. This disease appears to be as likely to disseminate to the mediastinal or hilar lymph nodes as to the pulmonary parenchyma, and it can simulate a primary bronchial tumor or lymphoma. PMID- 8424325 TI - Increased density of the azygos lobe on frontal chest radiographs simulating disease: CT findings in seven patients. AB - OBJECTIVE: Our objectives were to show that an opaque azygos lobe on the anteroposterior chest radiograph can be a normal finding and to investigate the causes of the increased density. MATERIALS AND METHODS: We prospectively examined 53 patients seen during a 3-year period in whom an azygos lobe was detected on chest radiographs. The size of the lobe was determined by the location of the trigone on chest radiographs. The trigone is the triangular area that marks the upper portion of the azygos fissure. If the trigone was located on the lateral aspect of the pulmonary apex, the azygos lobe was classified as type A (nine patients); if the trigone was situated at the midpoint of the cupula of the apex, the lobe was considered type B (20 patients); if the trigone was located on the medial face of the apex, the lobe was considered type C (24 patients). An opaque azygos lobe was defined as an increase of density suggestive of a pulmonary or mediastinal process. The findings on chest radiographs were compared with those on CT scans, which were obtained in all patients. RESULTS: Radiographs showed abnormally opaque azygos lobes in seven patients. Three cases were type B and the opaque lobe had a triangular aspect, suggesting pulmonary disease. Four cases were type C and the opacity had a convex aspect, suggesting a mediastinal mass. In all cases, CT scans showed a shallow azygos lobe and increased depth of the soft tissues of the mediastinum in front of the azygos lobe. This appearance was caused by tortuosity of the supraortic vessels in six cases and by a shadow from a normal thymus in one case. No pathologic process was discovered in any of the patients. CONCLUSION: We believe an opaque azygos lobe can be a normal finding, usually resulting from overlapping tortuous supraortic vessels. PMID- 8424326 TI - Ground-glass opacity of the lung parenchyma: a guide to analysis with high resolution CT. AB - Ground-glass opacity is a frequent but nonspecific finding on high-resolution CT scans of the lung parenchyma. The underlying abnormality is diverse; any condition that decreases the air content of the lung parenchyma without totally obliterating the alveoli can produce ground-glass opacity. These processes are not visible on high-resolution CT scans. However, in specific clinical settings, the information provided by high-resolution CT is considerable when the anatomic distribution and associated structural changes to the lung parenchyma are analyzed. This pictorial essay illustrates the pathologic basis of ground-glass opacity and provides a guide to the differential diagnosis of the disorders that can produce this appearance. PMID- 8424327 TI - Bronchiectasis: CT evaluation. AB - CT is the imaging method of choice after standard chest radiography for examining patients with suspected bronchiectasis. In most institutions throughout the world, CT has largely eliminated the need for bronchography in the diagnosis of bronchiectasis. Nonetheless, controversy persists concerning the overall accuracy of CT. In an effort to improve overall diagnostic accuracy, we review the wide range of CT appearances of this protean disorder, and emphasize potential problems and technical pitfalls that may arise in routine clinical imaging. PMID- 8424328 TI - Francis Henry Williams: the father of cardiac fluoroscopy. PMID- 8424329 TI - Epicardial lipoma: imaging findings. PMID- 8424330 TI - Tubular carcinoma of the breast: mammographic appearance. AB - OBJECTIVE: Tubular carcinoma of the breast is an uncommon malignant growth that can have subtle mammographic findings. Tubular carcinoma is usually found incidentally during screening mammography. It can be differentiated from conventional breast cancers by its smallness and the lack of palpable findings on physical examination of the breast. The mammographic findings in patients with pure tubular carcinoma (90% tubular formation or greater on histologic examination) have not been reported in the recent radiologic literature. MATERIALS AND METHODS: We reviewed the mammographic and sonographic findings and clinical histories of 13 patients with biopsy-proved pure tubular carcinoma. Findings on physical examination of the breast were normal in nine patients. RESULTS: On mammograms, 11 of the 13 tubular carcinomas appeared as small (1.7-cm diameter or less) spiculated masses. The average tumor diameter measured on mammograms was 0.8 cm in patients with normal results of physical examination of the breasts and 1.2 cm in patients with a palpable mass. In two patients, mammography showed suspicious microcalcifications or normal findings. Sonography was performed in three patients and results were normal. CONCLUSION: Tubular carcinoma of the breast manifests mammographically as a small spiculated mass. Because of their smallness, tubular carcinomas can be differentiated from other forms of infiltrating ductal carcinoma on mammograms. Tubular carcinomas are most frequently found incidentally at screening mammography. With the growing emphasis on screening mammography, an increase in the detection of this form of breast cancer can be expected. PMID- 8424331 TI - Mammographic findings in men with breast cancer. AB - OBJECTIVE: Our objective was to identify the mammographic findings of breast cancer in men. MATERIALS AND METHODS: The mammograms of 23 men with proved breast cancer (mean age, 63 years; range, 44-86 years) were retrospectively reviewed. Medical histories included gynecomastia in five, prior cancer in three, and radiation exposure in two. The most common signs were a mass in 13 and bloody nipple discharge in eight. RESULTS: Carcinoma was evident mammographically as an uncalcified mass in 17 patients (74%) and as a mass with microcalcifications in two patients (9%). Three tumors were not evident on mammograms, including one that was obscured by gynecomastia. Tumors were largely subareolar (14/17, 82%), and all were ductal cancers, including six pure intraductal carcinomas. CONCLUSION: Mammograms of men with breast cancer usually show an uncalcified subareolar mass, which may mimic or be obscured by gynecomastia. If calcifications are present, they may not have a pattern usually associated with malignancy. PMID- 8424332 TI - Quality assurance in mammography: status of residency education. AB - OBJECTIVE: The purpose of this study was to determine to what extent radiology residents are being trained in quality assurance procedures for mammography. MATERIALS AND METHODS: A telephone survey was conducted with 189 chief residents and 10 other senior residents from 199 of the 209 residency training programs in diagnostic radiology. RESULTS: Time spent on mammography rotations averaged 8 weeks. Only 10 residents (5%) were "very familiar" with the American College of Radiology (ACR) Mammography Accreditation Program; 72 (36%) were "not at all familiar" with it. Ninety-six (48%) knew that one technologist should be assigned quality control procedures. The majority did not know the recommended frequency for performing any of the five routine quality control procedures; only one knew the recommended frequencies for all five. Only twenty-seven (14%) knew the recommended maximum dose for a mammogram. Regarding biopsy yields and false negative results: 92 residents (46%) sat in on outcome evaluation sessions; 54 (27%) played active roles, looking up and tabulating results; and 53 (27%) did not participate at all. CONCLUSION: Although time spent on mammography rotations has increased substantially, quality assurance issues are still largely neglected. It may not be reasonable for radiology residents to have detailed instruction in quality assurance procedures for mammography, but they should be more familiar with the general issues involved and the procedures intended to correct the problem of the variable quality of mammography in this country. PMID- 8424333 TI - Appearance of oxidized cellulose (Surgicel) on postoperative CT scans: similarity to postoperative abscess. AB - OBJECTIVE: Oxidized regenerated cellulose (Surgicel), a sterile knitted fabric that causes thrombus formation because of its physical properties, is frequently used for intraoperative hemostasis. Unlike traditional surgical sponges, it is bioabsorbable and can be left in the surgical bed. On CT scans, the appearance of the retained oxidized cellulose can mimic that of an abscess. The purpose of this study was to describe the appearance of oxidized regenerated cellulose on postoperative CT scans so that an erroneous diagnosis of an abscess can be avoided. MATERIALS AND METHODS: We reviewed the CT examinations of five postoperative patients in whom oxidized regenerated cellulose had been used for surgical hemostasis. Operative reports and surgeons confirmed the use of oxidized cellulose. Four CT scans were abdominal examinations, and one was a head examination. RESULTS: In four cases, CT scans showed focal, linear collections of gas within masses with mixed attenuation in or near the operative site. No air fluid levels were present. In three patients, cultures of specimens obtained by aspiration were negative for pyogenic organisms. CONCLUSION: Retained oxidized cellulose can mimic an abscess on CT scans. Focal collections of air centrally located within a mass should alert the radiologist that oxidized cellulose may have been placed in the operative site, and an appropriate history should be sought. PMID- 8424334 TI - Primary ileocecal tuberculosis. PMID- 8424335 TI - Periportal low density on CT in patients with blunt trauma: association with elevated venous pressure. AB - OBJECTIVE: We postulated that the CT finding of periportal low density after acute blunt trauma of the abdomen or pelvis results from elevated central venous pressure caused by rapid expansion of intravascular volume during IV resuscitation or by other trauma-related pathologic changes. MATERIALS AND METHODS: CT scans and medical records of 58 patients, including 42 with CT evidence of hepatic injury and 16 with periportal low density without CT evidence of hepatic injury, were reviewed retrospectively to determine the extent of liver injury, the extent of periportal low density, and the ratio of the transverse diameter of the inferior vena cava to the transverse diameter of the aorta at the level of the right adrenal gland. The ratio, which served as an indirect measure of central venous pressure, was also measured in 20 randomly selected nontrauma patients with normal findings on abdominal CT scans. RESULTS: The ratios in 29 patients with CT evidence of liver injury without periportal low density were 0.6 2.0 (mean, 1.22), the ratios in 13 patients with diffuse (six) or focal (seven) periportal low density were 0.8-1.6 (mean, 1.31), and the ratios in 16 patients with periportal low density but no evidence of liver injury were 1.1-2.5 (mean, 1.36). The ratios were not statistically different among these three groups of patients. The ratios were 0.5-1.3 (mean, 0.86) for the 20 nontrauma control patients and were significantly lower than those in the trauma patients (p < or = .05). In all patients with periportal low density, the inferior vena cava remained distended at multiple levels through the abdomen without the variation in size expected from respiration, whereas a change in diameter was always observed on sequential CT sections in the nontrauma group. Among the 16 patients with periportal low density but no hepatic injury, a likely cause for elevated central venous pressure, excluding IV volume expansion, was observed in three. CONCLUSION: The CT finding of periportal low density after blunt abdominal trauma indicates acute elevation of central venous pressure and does not by itself signify hepatic parenchymal injury, as has been previously reported. PMID- 8424336 TI - Edward B. Singleton, 1992 gold medalist, Society for Pediatric Radiology. PMID- 8424337 TI - False-positive CT portography: correlation with pathologic findings. AB - OBJECTIVE: A retrospective study was performed to determine the causes of false positive diagnoses based on CT findings obtained during superior mesenteric arterial portography (CT portography) and to correlate the imaging characteristics of the incorrectly diagnosed lesions with their pathologic findings. MATERIALS AND METHODS: In a series of 52 patients who had CT portography before surgical exploration of the liver, eight had a total of 10 false-positive findings, yielding a false-positive diagnosis rate of 15%. In eight cases, the false-positive findings from CT portography were correlated with the histologic material obtained during partial hepatic resection or intraoperative biopsies at the corresponding sites. RESULTS: Of the 10 false positive findings on CT portograms, seven led to false-positive diagnoses of hepatic tumors. Conversely, three nontumorous false-positive findings were erroneously considered to be portal perfusion defects. The final diagnoses based on pathologic findings included focal fatty infiltration of the liver (three), cirrhosis (two), and portal perfusion defect (three). Two false-positive findings for which no histologic correlation was possible were considered to be portal perfusion defects on the basis of intraoperative findings. CONCLUSION: Recognition of false-positive findings is crucial in the preoperative evaluation of hepatic tumors because the findings may be interpreted as contraindications for surgery in patients who have operable tumors. The pathologic correlations show that several different entities can cause diagnoses based on CT portographic findings to be false-positive. However, differentiating between those entities remains a diagnostic challenge. PMID- 8424338 TI - Hepatic artery thrombosis in children with liver transplants: false-positive findings at Doppler sonography and arteriography in four patients. AB - OBJECTIVE: Patency of the hepatic artery in patients with liver transplants is evaluated with duplex Doppler sonography. In many centers, loss of an arterial waveform in the liver hilum is an indication for immediate arteriography to confirm the presence of hepatic artery thrombosis. We describe the findings in four children with liver transplants in whom occlusion of the graft artery was erroneously suggested by findings on duplex Doppler sonography and angiography. MATERIALS AND METHODS: We describe four patients 14-26 months old who had undergone liver transplant 9 days to 9 weeks earlier. The patients were critically ill with sepsis and hypotension. Duplex Doppler sonography was performed by interrogation of the hepatic artery in the liver hilum and parenchyma. When loss of an arterial waveform in the hilum was identified, angiography was performed immediately. Angiography consisted of aortography in all patients and selective celiac or superior mesenteric angiography in three patients. Autopsy was performed in all patients. RESULTS: Duplex Doppler sonography showed loss of arterial waveforms in the liver hilum in all patients; intrahepatic arterial waveforms were absent in three. The hepatic artery was not opacified at arteriography, but all patients had a patent hepatic artery at autopsy. Postmortem hepatic histology showed massive hepatic necrosis in three patients--necrosis without rejection in two and necrotizing vasculitis associated with severe rejection in one. The fourth patient had minor hepatic parenchymal injury. CONCLUSION: We conclude that failure to show flow in the hepatic artery with duplex Doppler sonography and arteriography is not necessarily indicative of arterial thrombosis. A low-flow nonocclusive phenomenon caused by massive hepatic necrosis or systemic hypotension may not be distinguishable from arterial occlusion. PMID- 8424339 TI - Three-dimensional localization of hepatic neoplasms with computer-generated scissurae recreated from axial CT and MR images. PMID- 8424340 TI - Evaluation of the pharyngeal airway in patients with sleep apnea: value of ultrafast MR imaging. AB - OBJECTIVE: Sleep apnea is often caused by obstruction of the pharyngeal airway. The goal of this study was to use ultrafast MR imaging to examine the pharyngeal airway in patients with sleep apnea and to evaluate the usefulness of this technique for localizing the site of obstruction. SUBJECTS AND METHODS: Fifteen patients with sleep apnea and five healthy volunteers underwent ultrafast MR imaging while awake and during sleep induced with hydroxyzine hydrochloride. Sequential midline sagittal images of the pharynx were obtained and displayed in the cine mode. RESULTS: Patients with sleep apnea were found to have sites of pharyngeal abnormality that were not present in healthy volunteers. Nine sites of narrowing in seven patients (47%) were detected with the patient awake; 21 sites of obstruction in 13 patients (87%) were diagnosed with the patient asleep. Six patients showed only one obstruction, and seven had several obstructions: five had obstructions at the velum palatinum and at the oropharynx; one had obstructions at the velum palatinum, oropharynx, and hypopharynx; one had obstructions at the velum palatinum and the hypopharynx. The sites of narrowing during wakefulness and the sites of obstruction during sleep were the same in only four (31%) of the patients with pharyngeal airway obstruction. CONCLUSION: Ultrafast MR imaging is useful for localizing the sites of pharyngeal airway obstruction in patients with sleep apnea. PMID- 8424341 TI - Colonic submucosal tumors: a new classification based on radiologic characteristics. AB - OBJECTIVE: Despite advances in gastrointestinal radiologic techniques, findings from barium enema studies cannot be used reliably to predict the histologic nature of a colonic submucosal tumor. Therefore, we generated a new and comprehensive classification system for all colonic submucosal tumors based on a correlation between their radiologic characteristics and their histopathologic features. MATERIALS AND METHODS: Between 1978 and 1990, 89 patients had 95 colonic submucosal tumors diagnosed on the basis of barium enema studies and confirmed by biopsy or surgery. Of these, 39 patients (44%) were symptomatic and 50 patients (56%) were asymptomatic. Single lesions (82 cases) and multiple lesions (12 cases of lymphoma and one case of blue rubber bleb nevus) were observed. The 95 lesions included 21 carcinoids, 27 malignant lymphomas, 17 lipomas, 10 lymphangiomas, six leiomyosarcomas, five leiomyomas, and nine others. We analyzed the radiologic findings and classified these lesions according to their appearance. RESULTS: All tumors were classified on the basis of their morphologic appearance into five types: (1) wide-based sessile lesion with gradually sloping margin and smooth surface (17 cases); (2) wide-based sessile lesion, more polypoid than the first type, with a smooth surface, with or without a central depression (49 cases); (3) wide-based sessile lesion with lobulated surface (six cases); (4) pedunculated lesion with smooth or granular surface (17 cases); (5) unclassified, not any of the previously mentioned types, may be diffusely stenotic or aneurysmal (six cases). The most common finding was the wide-based sessile lesion with a smooth surface and a possible central depression; next most common was the wide-based sessile lesion with gradually sloping margin and smooth surface; next was the pedunculated lesion with a smooth or granular surface. Eighty-six percent of the lesions were single; 14% were multiple. Histologic confirmation revealed mainly lymphoma (29%) and carcinoid (22%), of which the most common finding was a wide-based sessile lesion with a smooth surface, with or without a central depression. CONCLUSION: This new classification based on radiologic characteristics covers all varieties of colonic submucosal tumors. It can serve as a guideline for predicting a specific diagnosis of a colonic submucosal tumor on the basis of radiologic findings. PMID- 8424342 TI - CT in the diagnosis of primary aldosteronism: sensitivity in 29 patients. AB - OBJECTIVE: The most common cause of primary aldosteronism is a small aldosterone secreting adrenal adenoma. With improvements in CT technology, smaller adrenal lesions can now be detected. We reviewed our experience with 29 patients with primary aldosteronism to assess the sensitivity of CT in detecting aldosterone secreting adenomas. MATERIALS AND METHODS: The records of all patients with biochemically proved Conn's syndrome who were referred for adrenal CT between 1982 and 1991 were reviewed. The CT examinations of each of these 29 patients were reviewed for evidence of hyperplasia or an adenoma. The interpretations were correlated with subsequent adrenal venous sampling (20 patients) or surgery (17 patients). RESULTS: Fourteen of 17 aldosteronomas were detected on CT scans (sensitivity, 82%). Adrenal tumors were not seen on CT scans in any of the 12 patients with hyperplasia, although the glands appeared diffusely enlarged in only seven of these patients. In no case was an adrenal tumor seen on CT scans that was not found at surgery (positive predictive value, 100%). CONCLUSION: If CT scans of patients with Conn's syndrome show a focal mass, ipsilateral adrenalectomy can be performed with the expectation of cure. If no mass is found, adrenal venous sampling can be used to detect an adenoma not shown on CT. PMID- 8424343 TI - Safety and efficacy of kidney transplant biopsy: Tru-Cut needle vs sonographically guided Biopty gun. AB - OBJECTIVE: We evaluated the safety and efficacy of using a 14-gauge Tru-Cut needle without imaging guidance vs using an 18-gauge Biopty gun with sonographic guidance for percutaneous biopsy of kidney transplants. MATERIALS AND METHODS: We retrospectively analyzed data from 105 biopsies (in 68 patients) in which the Biopty gun with an 18-gauge needle was used and data from 100 biopsies (in 64 patients) in which a 14-gauge Tru-Cut needle was used. RESULTS: Significantly fewer major complications (p = .03) occurred when the Biopty gun was used (2%) vs when the Tru-Cut needle was used (10%). Two major complications (2%), both hematuria requiring transfusion, occurred in the 105 biopsies performed with the Biopty gun. Major complications occurred in 10 (10%) of the 100 biopsies done with the Tru-Cut needle: three obstructed allografts caused by blood clots, two episodes of hypovolemic shock, one case of shock and obstruction, and four intraperitoneal hemorrhages (one of which required nephrectomy). Biopsy specimens were adequate for histopathologic diagnosis in more than 98% of cases for both the Biopty gun and the Tru-Cut needle. CONCLUSION: We conclude that imaging guided percutaneous biopsy of renal allografts with a Biopty gun is as accurate as and safer than biopsy with the Tru-Cut needle. PMID- 8424344 TI - Pelvic fistulas: findings on MR images. PMID- 8424345 TI - Mayer-Rokitansky-Kuster-Hauser syndrome: distinction between two forms based on excretory urographic, sonographic, and laparoscopic findings. AB - OBJECTIVE: The purpose of this study was to discriminate typical (type A) from atypical (type B) Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome (congenital absence of vagina and uterus) and determine their association with renal anomalies and ovarian disease. MATERIALS AND METHODS: The excretory urographic, sonographic, and laparoscopic findings in 91 patients with MRKH syndrome were compared retrospectively. Symmetric muscular buds and fallopian tubes were diagnostic of type A, and asymmetric muscular buds or abnormally developed fallopian tubes were diagnostic of type B. RESULTS: On the basis of laparoscopic findings, type A was diagnosed in 40 patients (44%) and type B was diagnosed in 51 patients (56%). Renal anomalies were found in 34 (37%) of the 91 patients, all of whom had type B syndrome. Renal agenesis and a pelvic kidney were the most common findings in the upper part of the urinary tract. Ovarian abnormalities were seen in 14 patients (15%), all of whom had type B syndrome. Sonography did not allow discrimination between types A and B in patients with normal kidneys (17/51 = 33%), but it provided important information in patients with associated cyclic abdominal pain, in cases of diagnostic dilemma, and in patients with associated renal anomalies. CONCLUSION: Discrimination between type A and type B of MRKH syndrome is important because associated renal and ovarian abnormalities occur only in type B. Laparoscopy is still needed to discriminate between these two forms. Sonography is useful for diagnosing cyclic abdominal pain and associated renal anomalies. PMID- 8424346 TI - Acute fracture of the femoral neck: assessment of femoral head perfusion with gadopentetate dimeglumine-enhanced MR imaging. AB - OBJECTIVE: Evaluation of the perfusion and viability of the femoral head after fracture of the femoral neck is important because the outcome of conservative treatment or joint-preserving surgery is adversely affected by the development of capital osteonecrosis. We evaluated the use of MR imaging, before and after IV administration of gadopentetate dimeglumine, for assessing perfusion of the femoral head in 13 patients with acute fracture of the femoral neck. SUBJECTS AND METHODS: Multiecho (1600/30-240 [TR/TE]) MR images were obtained before contrast administration and gradient-echo (315/14, 90 degrees flip angle) MR images were obtained both before and after contrast administration. MR findings were correlated with findings on superselective digital subtraction angiograms of the vessels supplying the femoral head and with clinical-radiographic follow-up for at least 12 months. RESULTS: Digital subtraction angiography showed impaired blood supply to the femoral head in five patients. On contrast-enhanced MR images of these patients, the femoral head did not enhance and was lower in signal intensity than were the enhancing femoral shaft and neck distal to the fracture and the enhancing femoral head on the unaffected side. In the patients with persistent perfusion, contrast-enhanced MR images showed a uniform increase in signal intensity in the femoral shaft and neck as well as the femoral head; the femoral head on the fractured side showed contrast enhancement similar to that on the healthy side. CONCLUSION: These preliminary results indicate that contrast enhanced MR imaging may be useful for noninvasive evaluation of femoral head perfusion after fracture of the femoral neck. MR findings also may aid the clinician in deciding between joint-preserving therapy and hip arthroplasty. PMID- 8424347 TI - Meniscal tears of the knee: preliminary comparison of three-dimensional MR reconstruction with two-dimensional MR imaging and arthroscopy. AB - OBJECTIVE: A pilot study was performed to evaluate the usefulness of three dimensional (3D) MR reconstructions in detecting meniscal tears. MATERIALS AND METHODS: Reconstructions of 24 knee menisci were made by using data from standard two-dimensional (2D) MR images. The results were compared with findings on 2D MR images and at arthroscopy. RESULTS: All 17 arthroscopically proved meniscal tears were shown on the 3D reconstructions. Seven tears in five patients were shown on 3D reconstruction but not on 2D MR images, even on retrospective examination. Three of the seven were radial tears and four were horizontal tears. The seven missed tears were all located in the posterior horns. CONCLUSION: Three dimensional reconstructions are accurate for determining meniscal abnormalities and may be better than 2D imaging for evaluating the posterior horns of the menisci. Three-dimensional reconstructions may be indicated when 2D MR imaging shows no abnormality in patients in whom clinical findings strongly suggest a meniscal tear. PMID- 8424348 TI - Hepatic injury from blunt trauma in children: follow-up evaluation with CT. AB - OBJECTIVE: Because CT is now used to assess the extent of abdominal injury after blunt trauma, children with hepatic injury often can be treated nonsurgically. We used a CT grading system to evaluate the frequency of complications and the time course of healing in children with hepatic injury from blunt abdominal trauma. MATERIALS AND METHODS: Forty-five children with CT or surgical evidence of hepatic injury after blunt abdominal trauma were followed up clinically and with serial CT scans. Hepatic injuries were graded as mild (< 25% of one lobe injured), moderate (25-50% of one lobe injured), or severe (> 50% of one lobe injured). Physical activities were limited until healing was verified with follow up CT scans. RESULTS: All mild hepatic injuries (n = 12) appeared to have healed on follow-up CT studies 1 week to 11 months after the injury. Sixty-seven percent of moderate hepatic injuries (n = 19) showed complete healing on CT scans 1-3 months after injury, and 80% showed complete healing between 3.5 and 6 months. In all 14 children with severe hepatic injuries, residual lesions were seen on CT scans up to 8 months after injury. Residual lesions were seen in seven of 11 children with severe injuries who were reexamined 9-15 months after trauma. Despite this protracted course, no delayed hepatic complications occurred. No difference was seen between unenhanced and IV contrast-enhanced CT findings in 28 of 32 studies. IV contrast material improved the resolution of residual lesions in two cases, and two residual lesions were identified only on unenhanced CT scans. CONCLUSION: CT grading of acute hepatic injuries is useful for estimating the time course of healing. Although mild and moderate hepatic injuries heal relatively quickly, severe hepatic injuries take months to resolve. Follow-up CT scans to verify complete hepatic healing should be obtained 3 months after mild injuries, 3-6 months after moderate injuries, and 9 months after severe injuries. PMID- 8424349 TI - Obstructive vs nonobstructive dilatation of the renal collecting system in children: distinction with duplex sonography. AB - OBJECTIVE: The resistive index in cortical or arcuate renal arteries of children was calculated to determine whether obstructive uropathy could be differentiated from nonobstructive dilatation. SUBJECTS AND METHODS: Kidneys (n = 176) were prospectively evaluated by using duplex Doppler sonography in patients 3 days to 20 years old. Obstruction was confirmed by renography with furosemide and/or by surgery. RESULTS: The normal resistive index was 0.57 +/- 0.06 and the normal difference in resistive indexes between kidneys was 0.03 +/- 0.02 (n = 15). Abnormal values indicative of ureteropelvic junction obstruction were defined as the normal mean +/- 2 SD (i.e., a resistive index of > or = 0.70 plus a difference in resistive indexes between kidneys of > or = 0.08). Patients with unilateral dilatation and obstruction at the ureteropelvic junction (n = 20) had a mean resistive index of 0.77 +/- 0.05 and a difference in resistive indexes between kidneys of 0.16 +/- 0.05 (p < .001 compared with patients with normal kidneys and p < .001 compared with patients with unilateral dilatation without obstruction). Patients with unilateral dilatation but without obstruction (n = 16) had a mean resistive index of 0.63 +/- 0.06 and a difference between kidneys of 0.06 +/- 0.04 (values within normal limits). The positive and negative predictive values of the obstruction criteria for unilateral collecting system dilatation were 95% and 100%, respectively. After successful surgical correction of ureteropelvic junction obstruction (n = 29), patients had a normal mean resistive index of 0.61 +/- 0.05 and a normal difference between kidneys of 0.03 +/- 0.03. Five patients examined both before and after surgery showed a statistically significant drop in the resistive index of the obstructed kidney (0.75 +/- 0.03 to 0.65 +/- 0.05, p < .05) after surgery and a small rise in the resistive index of the contralateral kidney (0.56 +/- 0.04 to 0.63 +/- 0.04, p < .02). CONCLUSION: The resistive index appears to be an effective parameter for the evaluation and follow-up of unilateral obstructive or nonobstructive ureteropelvic junction dilatation in children. PMID- 8424350 TI - Renal calcifications in patients with autosomal recessive polycystic kidney disease: prevalence and cause. AB - OBJECTIVE: We investigated the prevalence of renal calcifications in children with autosomal recessive polycystic kidney disease and studied the metabolic changes that could cause this complication. SUBJECTS AND METHODS: Nine patients with known autosomal recessive polycystic kidney disease were examined with sonography and CT and screened for biochemical evidence of metabolic causes of nephrocalcinosis. RESULTS: CT showed bilateral renal calcifications in seven of the nine patients. The two patients without renal calcifications were less than 1 year old. Four patients had only a few calcifications and three patients had many. The severity of the renal calcifications correlated with the degree of kidney failure. All patients with kidney failure were found to have urine acidification defects. Hypocitraturia was present in all patients. CONCLUSION: Our results show that renal calcifications are common in older children with autosomal recessive polycystic kidney disease. Hypocitraturia and the urine acidification defect resulting from kidney failure are the leading factors in the pathogenesis of the calcifications. PMID- 8424351 TI - Gas enema for the reduction of intussusception: relationship between clinical signs and symptoms and outcome. AB - OBJECTIVE: The aim of this study was to establish the extent to which the clinical features of intussusception can be used to predict successful outcome of gas enema and to determine whether the nonsurgical management of intussusception in children can be improved by refining the criteria used to select patients for gas enema. SUBJECTS AND METHODS: Clinical data on 282 consecutive episodes of intussusception (255 patients) were collected prospectively from January 1987 to July 1991. Gas enema was performed in 273 episodes, in which the clinical signs and symptoms were studied by using logistic regression. Nine patients had primary surgery. RESULTS: Gas enema was successful in 216 (79%) of 273 enemas attempted. Fifty-seven patients had surgery after unsuccessful enema. Univariate analysis showed significant associations between successful enema and duration of signs and symptoms less than 12 hr, no rectal bleeding, absence of small-bowel obstruction, presence of a palpable mass, and normal hydration. Multivariate analysis showed that dehydration, small-bowel obstruction, and duration of signs and symptoms longer than 12 hr were significant predictors of unsuccessful enema; yet, in these groups the rate of success still justified attempted enema. Even in severe dehydration, the successful enema reduction rate was 31%. CONCLUSION: Our data suggest that although the factors identified had some predictive value in determining the outcome of attempted enema reduction, they could not be used to indicate patients in whom enema reduction should not be attempted. All patients with intussusception should have a gas enema if the absolute contraindications to enema (i.e., peritonitis or perforation) are absent. PMID- 8424352 TI - Bilateral persisting mesonephric ducts. PMID- 8424353 TI - Extragonadal yolk sac tumor: sonographic and CT findings. PMID- 8424354 TI - Characteristic MR findings in a neonate with incontinentia pigmenti. PMID- 8424355 TI - Partial anomalous pulmonary venous drainage of the left upper lobe vs duplication of the superior vena cava: distinction based on CT findings. AB - OBJECTIVE: Partial anomalous pulmonary venous drainage of the left upper lobe and duplication of the superior vena cava have similar appearances on CT scans. The purpose of this study was to review their appearances and provide guidelines for differentiating between them. MATERIALS AND METHODS: A review of the CT reports for the preceding 4 years disclosed seven patients whose original diagnosis was duplication of the superior vena cava and one patient whose diagnosis was partial anomalous pulmonary venous drainage of the left upper lobe. The 14 CT examinations of these eight patients were reviewed in order to observe the CT findings in each anomaly. RESULTS: In only five of the seven patients whose original diagnosis was duplication of the superior vena cava were CT findings compatible with that diagnosis. In the other two, CT findings were compatible with partial anomalous pulmonary venous drainage of the left upper lobe, as they were in the one patient with that as his original diagnosis. Two CT findings allow consistent differentiation. In duplication of the superior vena cava, two vessels can be seen anterior to the left main bronchus, whereas no vessels are present in this location in partial anomalous pulmonary venous drainage of the left upper lobe. Additionally, careful inspection reveals that the intraparenchymal veins in the left upper lobe drain to the normally positioned left superior pulmonary vein in duplication of the superior vena cava, whereas they drain to the anomalous vessel in partial anomalous pulmonary venous drainage of the left upper lobe. CONCLUSION: Careful analysis of the CT scans with particular attention to these two features allows differentiation between partial anomalous pulmonary venous drainage of the left upper lobe and duplication of the superior vena cava. PMID- 8424356 TI - Coronary artery fistula after cardiac transplantation: atypical location. PMID- 8424357 TI - Compression repair of a postcatheterization pseudoaneurysm of the brachial artery under sonographic guidance. PMID- 8424358 TI - Migration of the Simon nitinol vena cava filter to the chest. PMID- 8424359 TI - Parietal lobe abnormalities detected with MR in patients with infantile autism. AB - OBJECTIVE: Infantile autism is a neurologic disorder that severely disrupts the development of many higher cognitive functions. The most consistent abnormal neuroanatomic findings in autism are loss of Purkinje neurons in the posterior cerebellum as detected by autopsy studies and hypoplasia of the posterior cerebellar vermis and hemispheres as detected by in vivo neuroimaging. Evidence of developmental arrest has also been detected in limbic structures in autopsy studies of autistic patients with mental retardation. Neither in vivo neuroimaging nor autopsy studies of autistic persons have reported abnormalities in the cerebrum. Because the cerebrum mediates many higher cognitive functions, such as social communication, language, abstract reasoning, planning, and organization, that are known to be deficient in patients with autism, a closer examination of the neuroanatomy of the cerebrum in infantile autism is warranted. MATERIALS AND METHODS: MR images of 21 healthy autistic patients (6-32 years old) were mixed with MR images of control subjects and reviewed on four separate occasions by a neuroradiologist for any neuroanatomic abnormalities. Autism was diagnosed on the basis of criteria for autism as defined by the Diagnostic and Statistical Manual of Mental Disorders, and the autistic patients did not have any other concurrent neurologic disorders. To control for systematic bias in judging the type and location of abnormalities in the autistic population, three control groups were used: a normal control group of 12 subjects, a control group of 23 nonautistic patients with a variety of brain abnormalities for the first review, and another control group of 17 nonautistic patients for the second review. Control patients with brain abnormalities were selected from patients' files on the basis of MR findings of a variety of brain abnormalities. All MR images were coded for anonymity, randomly mixed, and examined by a neuroradiologist blinded to the purpose of the study and to the group membership of each subject. All normal and abnormal findings seen on the MR images of each subject were described on a standard form listing all major brain structures to ensure an examination of each structure in turn. To test for reliability, three subsequent reviews were performed by the same neuroradiologist. RESULTS: Parietal lobes were abnormal in appearance in 43% (9/21) of autistic patients. Cortical volume loss in the parietal lobes was seen in seven autistic patients; in four of these cases, cortical volume loss extended either into the adjacent superior frontal or occipital lobe. Additional abnormalities detected with MR in these nine patients included white matter volume loss in the parietal lobes (three patients) and thinning of the corpus callosum, especially along the posterior body (two patients). Abnormalities were bilateral. The mesial, lateral, and orbital regions of the frontal lobes; temporal lobes; limbic structures; basal ganglia; diencephalon; and brainstem were normal in all autistic patients. No abnormalities were found in the 12 normal control subjects. The control subjects with neurologic abnormalities had various abnormal findings consistent with their medical conditions. CONCLUSION: Our results indicate that the parietal lobes are reduced in volume in a portion of the autistic population. Possible origins for this localized cerebral abnormality include early-onset altered development and late-onset progressive atrophy. PMID- 8424360 TI - Acoustic schwannoma: MR findings in 84 tumors. AB - MR imaging is the study of choice for the examination of patients with suspected acoustic schwannoma, because of its high sensitivity, especially after the use of contrast material. This essay illustrates the common MR features of acoustic schwannomas as seen in a study of 84 tumors. We pay special attention to the role of MR imaging in the distinction between acoustic schwannoma and meningioma. PMID- 8424362 TI - Audible radiation monitors: the value in reducing radiation exposure to fluoroscopy personnel. PMID- 8424361 TI - Perfluorooctylbromide as a contrast agent for CT and sonography: preliminary clinical results. AB - OBJECTIVE: Perflubron emulsion is a bromine-based particulate contrast agent that is taken up selectively by the liver and spleen after IV injection. It does not leave the vascular space and also persists in the blood for a longer period than iodinated contrast agents do. We performed a preliminary study to determine the value of the IV perflubron emulsion as a contrast material for CT and sonography. SUBJECTS AND METHODS: To determine the safety and dose response of perflubron emulsion as an IV contrast agent for CT and sonography, 18 cancer patients, 14 of whom had hepatic metastases, were given 0.5-3.0 ml/kg of the emulsion. Sonography and CT of the liver and spleen were performed before and immediately after infusion and then again at 24 hr. RESULTS: A persistent increase in the density of blood, liver, and spleen was observed, where maximal enhancements of 55, 39, and 317 H, respectively, were achieved. Tumor conspicuity increased as metastases enhanced minimally (7 H or less). Peak enhancement of the liver and spleen was delayed to 24 hr with higher doses of perflubron; however, the immediate postinfusion scan allowed the distinction of vessels from hepatic lesions. Sonographically, an increase in tumor echogenicity relative to liver was observed in nine of the 14 subjects, allowing the detection of additional metastases in two. Splenic echogenicity in the near field increased relative to kidney; however, attenuation increased markedly at the time of peak concentration, limiting beam penetration. Similar but milder changes were observed in the liver. Adverse effects occurred in 14 of the subjects; these included lower back pain in six, delayed fever in eight, and malaise in three. CONCLUSION: Perflubron emulsion produced prolonged enhancement of blood, liver, and spleen and increased the conspicuity of liver metastases. However, the side effects encountered could limit its use to a selected population of patients. PMID- 8424363 TI - Manuscript peer review: general concepts and the AJR process. PMID- 8424364 TI - A surfeit of superfluous and redundant pleonasms. PMID- 8424365 TI - The AJR of the future: electronic publication and distribution. PMID- 8424366 TI - Unusual mammographic findings in a patient with partial lipodystrophy. PMID- 8424367 TI - Hidden costs of mobile mammography. PMID- 8424368 TI - The mammographic density of breast cancer. PMID- 8424369 TI - Pulmonary infarction: use of color Doppler sonography for diagnosis and assessment of reperfusion of the lung. PMID- 8424370 TI - Motion artifact simulating aortic dissection. PMID- 8424371 TI - Reflux parotid sialogram during barium swallow. PMID- 8424372 TI - Transjugular intrahepatic portosystemic shunt: the left internal jugular vein approach. PMID- 8424373 TI - "Tunnel sign" in transjugular intrahepatic portosystemic shunts. PMID- 8424374 TI - Intrahepatic portosystemic venous shunt in liver cirrhosis: is it congenital or acquired? PMID- 8424375 TI - Capsular retraction in hepatic tumors. PMID- 8424376 TI - Percutaneous transluminal angioplasty of inferior vena cava and hepatic veins for Budd-Chiari syndrome. PMID- 8424377 TI - A national survey of attitudes and practices of primary-care physicians relating to nutrition: strategies for enhancing the use of clinical nutrition in medical practice. AB - A nationwide mail survey was used to determine the degree to which primary-care physicians indicated that they practice the "core competencies" in clinical nutrition identified by Young et al (Am J Clin Nutr 1983;38:800-10). We also surveyed the nutrition-related attitudes of these physicians. Although the 3416 physicians who responded to the survey tended to report favorable attitudes toward using nutrition in their practice, these favorable attitudes were not consistent with their own reports of clinical performance. Neither the positive- or negative-attitude score correlated highly with the reported behavior-practice score. The clinical practices reported by those surveyed are well below the minimum level defined by the Young et al essential core competencies in clinical nutrition. The attitudes, practices, and demographic characteristics associated with the clinical performance variables suggest educational strategies for improving the competence of primary-care physicians and medical students in clinical nutrition. PMID- 8424378 TI - Effect of phenylpropanolamine on energy expenditure and weight loss in overweight women. AB - The effect of phenylpropanolamine (PPA), a noncatecholamine sympathomimetic weight-loss agent, on energy expenditure (EE) and substrate oxidation was measured in a respiratory chamber in 24 overweight women after 4 d of treatment (PPA or placebo) during weight maintenance and after 7 wk of treatment on a hypoenergetic diet (70% of measured baseline 24-h EE). Twelve women (37 +/- 2 y, 74 +/- 6 kg, 33 +/- 1% body fat) were randomly assigned to the PPA group [75 mg osmotic release oral system (OROS)-PPA/d] and 12 (mean +/- SEM: 38 +/- 2 y, 79 +/ 1 kg, 37 +/- 1% body fat) to the placebo group. Baseline measurements of 24-h EE (7849 +/- 226 vs 7834 +/- 142 kJ/d), basal metabolic rate (BMR) and 24-h respiratory quotient (RQ) were comparable between PPA and placebo groups. After 4 d of treatment, there was no significant effect of PPA on 24-h EE, BMR, and 24-h RQ compared with placebo. Over the 7-wk diet period, however, the PPA group (n = 8) had greater weight loss than the placebo group (n = 10): -5.0 +/- 0.5 vs -3.0 +/- 0.4 kg (P < 0.05). The changes in 24-h EE and 24-h RQ over the 7 wk were not different between the groups. We conclude that weight loss is enhanced by OROS PPA, but this change was not explained by changes in 24-h EE or 24-h RQ. The small number of subjects may have hindered detection of subtle differences in energy metabolism. PMID- 8424379 TI - Severe vs moderate energy restriction with and without exercise in the treatment of obesity: efficiency of weight loss. AB - Thirty obese women were randomly assigned to either 40% [severe energy restriction (SER)] or 70% [moderate energy restriction (MER)] of their maintenance energy requirements and to no exercise, aerobic exercise (walking), or aerobic exercise plus circuit weight training. Body composition by hydrostatic weighing and energy expenditure by indirect calorimetry were measured at 0, 3, and 6 mo. In addition, we developed a deficit-efficiency factor (DEF), calculated as body energy loss/dietary energy deficit, to attempt to quantify the effectiveness of the weight-reduction interventions. Subjects in the SER group lost more weight (mean +/- SE: 15.1 +/- 1.4 vs 10.8 +/- 1.0 kg), fat (11.7 +/- 1.1 vs 8.3 +/- 0.6 kg), and fat-free mass (2.8 +/- 0.3 vs 1.8 +/- 0.3 kg) than the MER group (P < or = 0.05). However, the overall DEF was greatest in the MER group (0.80 +/- 0.07) compared with the SER group (0.52 +/- 0.05; P < or = 0.01). Exercise had no significant effect. This study demonstrates that MER may offer an advantage over SER because it produces a greater energy loss relative to energy deficit. PMID- 8424380 TI - Low compliance with an iron-supplementation program: a study among pregnant women in Jakarta, Indonesia. AB - The efficiency of an established iron-supplementation program for pregnant women in Jakarta, Indonesia was investigated. Hemoglobin, serum ferritin, and packed cell volume (PCV) were measured at the start of the study and after 2 mo supplementation with 300 mg ferrous sulphate/d. The women (n = 45) were questioned about compliance and stool samples were checked for iron content to control for tablet intake. Twelve women dropped out. Prevalence of anemia (42%) did not decrease during the study period. Of the remaining 33 women, 64% (n = 21) claimed to have taken all iron tablets. This was only confirmed by positive stool tests in 12 women. Serum ferritin and PCV increased in women with positive stool tests (P < 0.05 and P < 0.01, respectively) after supplementation. It is concluded that compliance was low and that the iron dose needs to be increased. Supplementation programs need reliable monitoring and evaluation systems. PMID- 8424381 TI - Breast-fed infants are leaner than formula-fed infants at 1 y of age: the DARLING study. AB - Anthropometric indexes from 1 to 24 mo were compared between matched cohorts of infants either breast-fed (BF, n = 46) or formula-fed (FF, n = 41) until > or = 12 mo. Neither group received solid foods before 4 mo. Weight-for-length was significantly greater among FF infants from 7 to 24 mo. In both groups, skinfold thickness (triceps, biceps, subscapular, flank, and quadriceps) and estimated percent body fat (%FAT) increased rapidly during the first 6-8 mo and declined thereafter. At all sites except biceps, FF infants had larger skinfold thicknesses in later infancy (particularly 9-15 mo) than did BF infants; %FAT was significantly higher from 5 to 24 mo. Lower energy intake among BF infants explained the difference between groups. Maternal and infant fatness were positively correlated at 12-24 mo. Breast-milk lipid and energy concentration were unrelated to infant fatness. These results indicate that infants BF for > or = 12 mo are leaner than their FF counterparts. PMID- 8424382 TI - Dietary and anthropometric determinants of plasma lipoproteins during a long-term low-fat diet in healthy women. AB - Long-term (1 y) effects of dietary fat intake on lipoprotein metabolism were determined in 72 healthy women receiving either a 15%-fat diet (n = 34) or usual diet (n = 38). Every three months food records, weight, waist-hip ratio (W:H), percent body fat, fasting plasma triglyceride, cholesterol (C), high-density lipoprotein cholesterol (HDL-C), HDL2-C, and HDL3-C; apolipoprotein B and A-I, and postheparin lipoprotein lipase (LPL) and hepatic triglyceride lipase activities were determined. In one year, the low-fat-diet (LFD) group had 17% and the non-intervention-diet group had 36% dietary fat. The LFD group showed decreases in cholesterol: 7% TC, 13% low-density lipoprotein (LDL), and 8% HDL. Apolipoprotein A-I, decreased early. Apolipoprotein B did not change. Plasma triglyceride correlated with weight. Percent body fat and W:H correlated with the total and LDL-C. Changes in HDL-C and/or HDL2-C and LPL correlated directly with the changes in dietary fat and inversely with dietary carbohydrate. Changes in total-C or LDL-C correlated with the changes in weight and W:H, but not with the changes in nutrient intake. PMID- 8424383 TI - Dietary modification of omega 6 fatty acid intake and its effect on urinary eicosanoid excretion. AB - A group of women were fed two separate diets in a crossover study and urinary eicosanoids were quantified. One diet contained 3.1% of total energy (en%) as polyunsaturated fatty acids (3.0 en% linoleic acid) and the other contained 8.4 en% polyunsaturated fatty acids (8.3 en% linoleic acid). Carbohydrate replaced fat in the low-polyunsaturated-fat diet. No changes were observed in the urinary excretion of 6-oxo-prostaglandin F1 alpha, its 2,3-dinor metabolite or thromboxane B2 by subjects on either of the diets. Urinary 2,3-dinor-thromboxane B2 excretion was lower (206.5 ng/24 h) when subjects were fed the high-omega 6 polyunsaturated fatty acid diet when compared with the lower-omega 6 polyunsaturated fatty acid diet (275.3 ng/24 h). Conversely, urinary prostaglandin E2 was higher (139.2 ng/g creatinine) during the higher-omega 6 polyunsaturated fatty acid diet when compared with the lower-omega 6 polyunsaturated fatty acid diet (94.4 ng/g creatinine). PMID- 8424384 TI - An isotopic study of the effect of dietary carbohydrate on the metabolic fate of dietary leucine and phenylalanine. AB - The fate of dietary leucine and phenylalanine was studied in five healthy, young adult men, by using a dual, stable isotope-tracer infusion approach to estimate amino acid fluxes, splanchnic (Sp) uptake, and dietary of absorbed amino acid to the peripheral circulation. Subject received two, 4-h tracer infusions of [1 13C]leucine and [15N]phenylalanine infused through a feeding tube placed in the duodenum, and [5,5,5-2H3]leucine, [ring-2H5]phenylalanine, and [6,6-2H2]glucose infused simultaneously by vein. In one experiment subjects received an amino acid mixture (83 mg amino acid.kg-1.h-1) via the feeding tube and in the other experiment amino acids were supplied with carbohydrate (CHO) (167 mg.kg-1.h-1). Sp uptake of dietary leucine decreased with added dietary CHO (29% of ingested leucine for amino acids alone vs 20% with CHO; P < 0.05) but was not different for phenylalanine (P > 0.05). Addition of CHO decreased both release of leucine via protein breakdown and leucine oxidation and increased body leucine balance (P < 0.05). PMID- 8424385 TI - Beta-carotene accumulation in serum and skin. AB - The accumulation of beta-carotene in serum and skin was evaluated in human volunteers. A single 51-mg dose of beta-carotene given in the absence of dietary fat resulted in no detectable change in serum beta-carotene. The same dose administered with 200 g fat increased serum beta-carotene 2.5-fold at 40 h. Similarly, administering beta-carotene daily in three divided doses with meals raised the serum beta-carotene concentration three times as high compared with the same total dose administered once a day; both regimens had the same time constant for serum accumulation; 9-10 d. Remittance measurements of skin color demonstrated that the accumulation of beta-carotene in skin was delayed by up to 2 wk compared with serum accumulation. These data indicate that beta-carotene absorption requires dietary fat and is enhanced by administering with meals but there is a long time constant for serum (10 d) and tissue (several weeks) accumulation. PMID- 8424386 TI - Comparison of micronutrient intake measured by a dietary questionnaire and biochemical indicators of micronutrient status. AB - We compared the intake of 12 micronutrients as reported on a semiquantitative food frequency questionnaire with corresponding biochemical indicators of nutrient status in a sample of 57 males and 82 females aged 40-83 y. Age-, sex and energy-adjusted correlation coefficients ranged from near zero for thiamin, vitamin A, and zinc to 0.63 for folate. Correlation coefficients between intake and the biochemical measures were > 0.30 for carotenoids, vitamin D, vitamin E, vitamin B-12, folate, and vitamin C. Differences of 50% or more were observed between extreme quartiles of intake for mean plasma concentrations of folate, vitamin B-12, and vitamin C. Excluding nutrient supplement users generally reduced the correlations. These data demonstrate that food frequency questionnaires can provide valid information on intake for a number of micronutrients. PMID- 8424387 TI - Effect of bovine-hemoglobin-fortified cookies on iron status of schoolchildren: a nationwide program in Chile. AB - The Chilean School Lunch program, which serves one million children nationwide, was supplied with three 10-g cookies fortified with 6% bovine hemoglobin concentrate, designed to provide 1 mg bioavailable iron per day. A survey of 1000 children was performed after 3 y. Significant differences in hemoglobin concentrations were found in the children from the fortified vs the nonfortified province (P < 0.01). Low serum ferritin values were also significantly more prevalent in the nonfortified group. The effect was evident despite the very low prevalence of anemia in both the fortified and the unfortified school groups. Heme-iron-fortified cookies are a feasible and effective way to improve the iron status of school-age children. In regions of high prevalence of iron-deficiency anemia, the effect of a heme-fortified cookie program should be even more important. PMID- 8424388 TI - Trace element intakes and dietary phytate/Zn and Ca x phytate/Zn millimolar ratios of periurban Guatemalan women during the third trimester of pregnancy. AB - Repeated 24-h recalls (9-14/subject) were conducted on 52 periurban Guatemalan pregnant women aged 25 +/- 5 y (means +/- SD). Intakes of energy, protein, calcium, zinc, copper, manganese, nonstarch polysaccharide (NSP), phytate, and millimolar ratios of phytate to zinc and (calcium x phytate) to zinc were calculated from food-composition values on the basis of chemical analysis and the literature. Mean (+/- SD) daily intakes were as follows: energy 8694 +/- 1674 kJ, protein 63.0 +/- 13.3 g, calcium 727 +/- 163, zinc 11.3 +/- 2.7, copper 1.3 +/- 0.3, manganese 2.8 +/- 0.6, phytate 2254 +/- 773 mg/d, NSP 26.6 +/- 6.9 g, phytate/zinc 18.8 +/- 4.2, (calcium x phytate)/zinc 706 +/- 21 mmol/MJ. Ninety four percent had zinc intakes below the recommendations (15 mg) of WHO and the US recommended dietary allowances, assuming 20% absorption. Tortillas were a major source of zinc (46%), copper (20%), manganese (23%), calcium (39%), phytate (68%), and NSP (50%); 19% zinc from flesh foods. Thirty-eight percent had phytate zinc ratios > 20; 94% had millimolar ratios of (calcium x phytate) to zinc per MJ > or = 22. The high prevalence of millimolar ratios of phytate to zinc and (calcium x phytate) to zinc per MJ above 20 and 22, respectively, may compromise zinc nutriture. PMID- 8424389 TI - Recovery of [13C]bicarbonate as respiratory 13CO2 in mechanically ventilated patients. AB - Measurement of the nutrient oxidation rate with 13C as a tracer requires knowledge of the value of its coefficient of fractional recovery in the expired gas (FR). We measured FR in nine intensive care patients who were mechanically ventilated and received total parenteral nutrition. NaH13CO3 was administered at a priming dose (3.75 mumol.kg-1.min-1) followed by a continuous infusion (0.05 mumol.kg-1.min-1). Metabolic rate and pulmonary carbon dioxide elimination (VCO2) were measured by using a mass-spectrometer system. The 13C-12C ratio was measured in the expired gas with an isotopic-ratio mass spectrometer and FR was calculated by using standard equations. The average value of FR was 0.899 +/- 0.026 (means +/- SE) and remained stable for each patient on 2 consecutive days. Between patients, the coefficient of variation of FR was 8.6%. Metabolic rate was the only physiological factor found to affect the FR value. PMID- 8424390 TI - Relative effects of dietary saturated, monounsaturated, and polyunsaturated fatty acids on cardiac arrhythmias in rats. AB - This study compared monounsaturated oleic acid with n-6 and n-3 polyunsaturated fatty acids (PUFAs) for their ability to modify the vulnerability to cardiac arrhythmias during ischemia or reperfusion in rats. Replacement of saturated animal fat in the diet with oleic acid-rich olive oil did not significantly alter the incidence of ventricular fibrillation or other cardiac arrhythmias. Replacement with either n-6-rich sunflower seed oil or n-3-rich fish oil reduced the incidence and severity of arrhythmias occurring in ischemia. The fish oil significantly reduced reperfusion arrhythmias independently of antecedent ischemic arrhythmias. Fatal ventricular fibrillation was significantly reduced by n-6 (8%; n = 25) and n-3 (0%; n = 24) PUFA but not by monounsaturates (36%; n = 25) compared with saturated fat (42%; n = 24). The results suggest that dietary replacement of saturated fats by n-6 and especially n-3 PUFA but not monounsaturated fatty acids can reduce the likelihood of an ischemic event leading to sudden cardiac death. PMID- 8424391 TI - Plasma ascorbic acid concentrations relate inversely to blood pressure in human subjects. AB - This study relates antioxidant status and blood pressure (BP) in 168 healthy residents of Augusta, GA, following usual diets. BP ranges were systolic (S) 84 152, mean 112 +/- 1 mm Hg, and diastolic (D) 52-96, mean 72 +/- 1 mm Hg. Plasma concentrations of ascorbic acid (AA) were significantly inversely related to SBP (r = -0.18, P < 0.05) and DBP (r = -0.20, P < 0.01); with regression equations SBP vs AA = -0.083C + 116 and DBP = -0.077C + 76. Highest and lowest quintiles of AA differed significantly in mean SBP (108 +/- 2, 113 +/- 2 mm Hg) and DBP (69 +/ 1, 74 +/- 2), P < 0.05. Plasma AA concentrations were significantly lower in the smokers. By deleting smokers, the inverse relations of SBP and DBP with plasma AA and the slopes of the equation were enhanced. Plasma selenium, alpha-tocopherol, alpha-tocopherol:cholesterol ratio, retinol and taurine were not related to BP; whereas male gender, body mass index, body fat distribution, plasma cholesterol, low density lipoprotein cholesterol, and triglycerides correlated. PMID- 8424392 TI - Effect of gluten-free diet on bone mineral content in growing patients with celiac disease. AB - Osteoporosis is a complication of celiac disease in adulthood, but little is known about the influence of the disease on bone mineralization in children. In the present study we evaluated radial bone mineral content (BMC) in celiac children and adolescents at diagnosis and after they consumed a gluten-free diet (GFD). The BMC values of 33 celiac patients at diagnosis were significantly lower than those of 255 control subjects (P < 0.001). There was no difference between diabetic and non-diabetic celiac patients. In 14 patients the BMC increased significantly (P < 0.05, ANCOVA) after 1.28 y of GFD. In these patients the mean annual BMC increment was 0.07 g/cm, significantly greater (P < 0.05) than the increment of normal growing children (0.05 g.cm-1.y-1). Our data indicate that although osteoporosis complicates celiac disease during childhood and adolescence, GFD alone is able to remarkably improve bone mineralization. PMID- 8424393 TI - Effects of monounsaturate-enriched diets on plasma lipoproteins. PMID- 8424394 TI - Specific dietary fatty acids in predicting plasma cholesterol. PMID- 8424395 TI - Fatty acids and liquid-formula diets in cholesterol studies. PMID- 8424396 TI - A randomized trial of mitomycin-C (M) versus mitomycin-C plus high-dose medroxyprogesterone acetate (MMPA) in the treatment of patients with advanced breast cancer. AB - Seventy-six women with previously treated breast cancer were randomized to receive mitomycin (M) or M plus high-dose oral medroxyprogesterone acetate (MMPA). Patients were balanced with respect to age, performance status, hormone receptor status, previous treatment, and number of metastatic sites. There were more patients with visceral metastases in the M arm of the study. Side effects were tolerable and not significantly different for the two regimens. No life threatening toxicity occurred. Objective response was documented in 4 of 37 patients on M and 11 of 39 on MMPA. On M the median time to treatment failure (TTF) was 3 months, and median survival was 7.8 months. On MMPA the median TTF was 4.4 months, and median survival was 9.7 months. There was a tendency for higher response and longer TTF and survival on MMPA, but statistical significance was not reached (p = 0.09). PMID- 8424397 TI - A phase II trial of vincristine in advanced or recurrent endometrial carcinoma. A Gynecologic Oncology Group Study. AB - Thirty-three evaluable patients who had not received prior chemotherapy were entered on a study of vincristine therapy for advanced or recurrent endometrial carcinoma. Vincristine 1.4 mg/m2 was given weekly as an i.v. bolus for 4 weeks and then every other week. There was one complete response (CR) lasting 5 months. Five patients had partial responses (PR) lasting 3-18 months. The CR+PR rate was 18% (95% confidence interval for CR+PR was 7-36%). Thirteen patients (38%) had stable disease from 2-28 months, and 14 had progressive disease. The major toxicity was neurological, with 11 patients having grade 2 or 3 peripheral neuropathy. Vincristine at this dose and schedule has modest activity, but troublesome toxicity in advanced or recurrent endometrial carcinoma. PMID- 8424398 TI - The frequency of red cell transfusion for anemia in patients receiving chemotherapy. A retrospective cohort study. AB - The objective of this retrospective cohort study was to determine the frequency of red cell transfusion for anemia and risk factors. It was conducted at a regional cancer center and host hospital. All patients receiving chemotherapy in 1989 were studied. Complete data were available on 381; 26 were excluded. Age, diagnosis, baseline and nadir hemoglobin, transfusion history, chemotherapy regimen, and prior treatment were abstracted from the cancer center chart and hospital medical records. Transfusion for anemia was required in 18% of all patients with solid tumors; those with lung cancer had the highest rate (34%). Patients with anemia who entered chemotherapy were more likely to be transfused. Therefore, patients with leukemia, lung cancer, and/or prior anemia have higher transfusion rates and may benefit from such therapies as recombinant human erythropoietin. PMID- 8424399 TI - Cancer of unknown primary. A retrospective study based on 109 patients. AB - Cases of 109 patients with metastases from cancer of unknown primary (CUP) were retrospectively analyzed. The highest survival rates were observed in patients with high performance status and metastases limited to the cervical lymph nodes. With other metastatic presentations, the survival was generally very short. Tumor markers did not demonstrate diagnostic value. The primary tumor was revealed by autopsy in 43 of 64 cases. Lungs were the most common primary site, followed by the gastrointestinal tract. Extensive clinical investigations during the initial diagnostic period were, as a rule, fruitless. Patients with CUP require efficacious treatment, but need to be investigated with adequate measures. PMID- 8424400 TI - Minor salivary gland tumors of the head and neck: treatment strategies and prognosis. AB - Between 1961 and 1985, 117 patients with malignant tumors of the minor salivary glands of the upper aerodigestive tract were treated with curative intent at the University of California, Los Angeles (UCLA). The length of follow-up ranged from 24-225 months, with a median of 38 months. The most common site of origin was the oral cavity (65 cases), and the most common histology was adenoid cystic carcinoma (69 cases). Tumor size, histology, and site were important prognostic factors. For oral cavity lesions, small tumors were well controlled with resection alone (25 of 26) or local excision, followed by irradiation (7 of 7). For advanced tumors arising from the paranasal sinuses and pharynx, the control rates were 43% (15 of 35) and 29% (5 of 17), respectively. Ninety-three percent (27 of 29) of patients with mucoepidermoid carcinoma had no evidence of disease at last follow-up versus 55% (38 of 69) of patients with adenoid cystic carcinoma and 56% (10 of 18) of those with adenocarcinoma. It appears that resection with good margin is adequate treatment for small lesions. Large, poorly differentiated tumors require a combined approach: surgery and radiation therapy. PMID- 8424401 TI - Intra-arterial hepatic treatment with carboplatin (CBDCA) and 5-fluorouracil (5 FU) in metastases from colorectal carcinoma. AB - Thirty-one patients with hepatic metastases from colorectal carcinoma were treated with carboplatin (CBDCA), 55 mg/m2, given in a 4-hour intra-arterial infusion daily for 5 days, and 5-fluorouracil, 900 mg/m2, given in a 20-hour intra-arterial infusion daily for 5 days. Cycles were administered every 5 weeks. Objective responses were observed in 16 (51.6%) patients (5 complete and 11 partial responses). Another 13 patients maintained stable disease, and 2 patients rapidly progressed. The overall median survival was 23.5 months. The 16 patients with objective response had a median survival of 26.5 months. In this series, no evidence of biliary sclerosis, cholecystitis, chemical hepatitis, or myelosuppression was observed. Complications of drug delivery system were observed in 14 (45.16%) patients. In conclusion, intra-arterial hepatic chemotherapy with CBDCA-5FU was associated with a modest benefit, expressed in good quality responses and extended survival of approximately 2 years in about half of the treated patients. PMID- 8424402 TI - A retrospective review of nodal treatment for vulvar cancer. AB - From 1983 to 1990, 42 clinical N0, N1 patients with invasive squamous cell carcinoma of the vulva underwent surgery for the primary cancer, followed by nonrandomized assignment to either surgery or radiation therapy for nodal management. This is a retrospective analysis reviewing treatment outcome and complications of inguinofemoral dissection versus photon irradiation. Group I (N = 24) underwent either bilateral or unilateral inguinofemoral dissection; Group II (N = 18) underwent bilateral groin irradiation. The 3-year disease-free survival was 84.5% and 79.7% for Group I and II (p = 0.74). The nodal failure rate at 3 years was not significantly different. The postoperative complications in the surgically treated nodes were: 17% seromas, 46% wound infection, and 71% wound separation. Of those patients developing either an infection or wound separation, 41% required daily wound care for more than 2 weeks by a visiting nurse after discharge. Only 1 of 18 (6%) irradiated patients developed a clinically significant skin reaction. The median time for complete skin/wound healing was 8 weeks (range 4-24 weeks) in Group I and 2 weeks (range 2-6 weeks) in Group II. Late postoperative complications in the surgically treated patients included meralgia paresthetica (8%) and lymphedema (12%); 1 (6%) Group II patient developed lymphedema. Based on this limited retrospective review, we conclude that inguinofemoral radiation achieves reasonable local control and may represent an alternative to surgery in selected patients. The acute and delayed morbidity of lymphadenectomy exceeds that of irradiation. PMID- 8424403 TI - The toxicity of daily inhaled amphotericin B. AB - Inhaled amphotericin was administered to 29 patients with prolonged neutropenia and toxicity was analyzed. Treatment consisted of 30 mg of amphotericin B administered by nebulizer once daily via either a hand-held nebulizer or face mask. The mean duration of treatment was 16 days. Toxicity was minimal and patient had significant toxic reaction to the inhaled medication. This study documents that nebulized amphotericin B has less than 10% incidence of severe toxicity with 95% confidence level. Guidelines for future trials and use are suggested. PMID- 8424405 TI - The use of cyclophosphamide, methotrexate, and 5-fluorouracil in the treatment of Merkel cell carcinoma. AB - Five patients with advanced Merkel cell carcinoma (MCC) are described. Four patients with regional lymph node involvement and one with disseminated skin metastases were treated with systemic chemotherapy, including cyclophosphamide, methotrexate, and 5-fluorouracil (CMF). The patients received a median of six cycles of CMF (range: 2 to 6), and chemotherapy was well tolerated. Four complete and one partial response were noted. Three patients are alive and are disease free at 5, 12, and 37 months from the onset of CMF chemotherapy. Two patients died from disseminated metastatic disease at 3 and 24 months from the onset of chemotherapy. CMF chemotherapy appears to be an active regimen in the treatment of locally advanced MCC. Further experience with this combination is warranted. PMID- 8424404 TI - A phase II trial of Didemnin B (NSC #335319) in patients with previously treated epithelial ovarian cancer. A Gynecologic Oncology Group Study. AB - Sixteen evaluable patients with advanced or recurrent epithelial ovarian carcinoma following progression on combination chemotherapy were treated with 4.2 mg/m2 of Didemnin B (NSC #325319) intravenously every 28 days until progression of disease. All patients had prior cisplatin-containing chemotherapy. There were no responders. Seven patients had stable disease (43.7%) and nine (53.3%) had increasing disease. The toxicities were significant, consisting of nausea and vomiting in seven patients (44%), one grade 3 hepatic toxicity, and three instances of grade 4 toxicities (1 leukopenia, 1 GI, and 1 GU). When used with this schedule Didemnin B is ineffective in patients with previously treated epithelial ovarian cancer. PMID- 8424406 TI - Impact of irradiation of residual breast on adjuvant chemotherapy dose intensity. AB - The chemotherapy dose intensity for breast cancer has been recently considered to be an important factor in determining clinical outcome. Many trials have shown routine postoperative irradiation to have a detrimental effect on the delivered dose of adjuvant chemotherapy. In this paper the impact of radiotherapy on the dose intensity of adjuvant chemo- or hormonotherapy was evaluated in 237 breast cancer patients. 177 patients had radical mastectomy and 60 quadrantectomy followed by radiotherapy. Chemotherapy comprised 6 cycles of FEC (5-fluorouracil, epidoxorubicin, cyclophosphamide) alternated to 6 cycles of CMF (cyclophosphamide, methotrexate, 5-fluorouracil)+tamoxifen for 5 years. There were no significant differences in the adjuvant chemotherapy dose intensity in the mastectomy group compared to the radiotherapy group. PMID- 8424407 TI - Intraoperative and external preoperative radiotherapy in invasive bladder cancer: effect of neoadjuvant chemotherapy in tumor downstaging. AB - Absence of residual cancer (pT0) in the cystectomy specimen was evaluated in patients with invasive bladder cancer treated with intraoperative (IORT) (15 Gy) and preoperative external beam radiotherapy (EBR) (46 Gy/5 weeks) with or without neoadjuvant chemotherapy. The overall pT0 rate was 68% (67% and 70% in patients with or without neoadjuvant chemotherapy, respectively). The tolerance to the program was acceptable in both groups. It is concluded that intense, combined modality treatment is feasible in bladder cancer patients, and the addition of neoadjuvant chemotherapy does not increase the morbidity. Preliminary results on disease-free survival are encouraging. PMID- 8424408 TI - The use of intraoperative electron beam radiation therapy in the treatment of para-aortic metastases from gynecologic tumors: a pilot study. AB - The abdomens of 16 patients with para-aortic nodal metastases were explored in the Clement O. Miniger Radiation Oncology Center's (COMROC) intraoperative radiation therapy suite. At the time of initial diagnosis 8 patients were found to have para-aortic metastases, and 8 developed para-aortic disease after completion of primary treatment. The median age of patients in the "initial group" was 37.5 years: 7 of these 8 patients had radiation only, and 1 patient had radiation and chemotherapy as initial treatment. Bulk disease in the para aortic region was noted in 2 patients, microscopic disease in 5 patients, and prophylactic radiation was given to the para-aortic area in 1 patient. Ten para aortic fields in 8 patients were treated with a median dose of 20 (range 10-25) Gy with 6-18 MeV electrons. Median survival from time of diagnosis was 31 (range 9-59) months and 27 (range 8-56) months from the date of IOEBRT. The median time to recurrence after IOEBRT was 8 (range 4-32) months. One patient is alive without disease 59 months and 1 is alive with disease 44 months from IOEBRT, with neither patient failing in the treated area. Eight patients were treated for recurrent disease in the para-aortic area: 5 cervical primaries, 2 endometrial adenocarcinoma primaries, and 1 uterine sarcoma. The median age was 60.5 (range 45-70) years. Median time to recurrence from initial diagnosis was 20 (range 7 37) months. In seven fields 6 of 8 patients received IOEBRT, median dose 20 (range 10-25) Gy. After IOEBRT, 4 of 6 patients received 45 Gy external beam irradiation. Median survival from IOEBRT was 9 (range 3-23) months, 37.5 (range 11-56) months from diagnosis. One patient is alive without disease at 11 months and one is alive with disease 19 months after IOEBRT. Toxicity of the IOEBRT is discussed. PMID- 8424409 TI - Treatment of indolent non-Hodgkin's lymphoma localized in the head and neck. AB - A total of 25 patients with indolent non-Hodgkin's lymphoma localized in the head and neck region was analyzed. Of these, 21 patients were classified as clinical Stage I, and 4 as Stage II. In 22 patients the extranodal site was involved, and it was a remarkable characteristic of this series of patients. Overall treatment results were as follows: survival rate 100% at 5 years, and relapse-free survival rate 81% at 2 and 5 years. There was only 1 death at 6 years and 2 months after treatment, and 4 relapses occurred by 2 years after treatment. Relapse was observed in 3 of 8 patients with follicular mixed/follicular medium type, which was in contrast to the other indolent histologies (SL, DSCl, ILL) with very few relapses. PMID- 8424411 TI - Outcome of care. Complications from radiation therapy treatment. AB - This study was undertaken to examine radiation treatment complications. From September 1987 through December 1989, 29,380 patients were screened at their follow-up visits for possible radiation complications. Of these patients, 1,380 were singled out for further study. These 1,380 charts were examined by a radiation oncologist and physicist to determine if there was a radiation complication, the severity of the complication, and whether a calculation or setup error could account for the complications. Of the 1,380 patients studied, 178 (3% of new patients treated each year) were determined to have radiation complications. These complications were divided into four categories of severity and entered into our computerized tumor registry as follows: complication type R1, complete recovery from symptoms, n = 59; R2, injury requiring medications to control injury, n = 104; S1, surgical intervention for one organ, n = 12; and S2, surgical intervention for two organs, n = 3. We believe that a baseline complication rate of 5% is acceptable in radiation oncology practices. However, the examination and documentation of the outcome of care in the form of radiation complications can help improve patient care and keep the radiation oncologist abreast of treatment outcome trends in the department. PMID- 8424410 TI - Phase II trial of a single intravenous dose of ondansetron in patients receiving cisplatin > or = 100 mg/m2. AB - Ondansteron, a serotonin antagonist antiemetic, controls emesis caused by cisplatin when given as multiple 0.15 mg/kg dosages in a variety of administration schedules. This trial evaluated a single 0.45 mg/kg i.v. dose of ondansetron in 21 cancer patients receiving cisplatin at doses > or = 100 mg/m2 as initial chemotherapy. Twenty-five percent of patients had no emetic episodes (95% confidence interval: 9-49%), and 45% had two or fewer emetic episodes (95% confidence interval: 23-69%). No significant adverse effects were seen. In prior trials at this institution in similarly treated patients, ondansetron given in three divided doses yielded a 42% no emesis rate. In this trial a single i.v. 0.45 mg/kg dose of ondansetron prevented emesis in 25% of patients receiving initial cisplatin > or = 100 mg/m2. For this dose of cisplatin and ondansetron, the observed emesis complete control rate was lower than that previously seen using multiple dose schedules. PMID- 8424412 TI - Metastatic breast carcinoma mimicking an acute paronychia of the great toe: case report and review of subungual metastases. PMID- 8424413 TI - High-dose carboplatin and mitoxantrone with autologous bone marrow support in the treatment of advanced breast cancer. AB - Ten patients with Stage II (four) and Stage IV (six) breast cancer were enrolled in a trial of conventional-dose induction therapy followed by high-dose therapy with autologous bone marrow support. Cyclophosphamide, methotrexate, and 5 fluorouracil were given to best response or five courses (Stage II). Those patients without progression were eligible for the high-dose portion of the protocol, which consisted of carboplatin 1,500 mg/m2 and mitoxantrone at either 40 mg/m2 (first five patients) or 50 mg/m2. Two patients did not receive the high dose portion of the treatment due to progression on induction therapy (one) and insurance refusal (one). Of the remaining eight patients who completed the high dose portion of the protocol, three were Stage II, of whom one died of transplant related complications, one progressed, and one is alive and free of disease 24 months after therapy. Of the five Stage IV patients, two achieved a partial remission, one of whom died of progressive disease 1 year after therapy and the other died of BMT-related complications; of the other three Stage IV patients, one had stable disease and died at +9 months, one with progression died at +3 months, and one died of BMT-related causes. Overall, three patients died of infectious complications, with two having alpha streptococcal septic shock syndrome. PMID- 8424414 TI - Nutcracker esophagus: an idea whose time has gone? PMID- 8424415 TI - The current role of erythromycin in the clinical management of gastric emptying disorders. PMID- 8424416 TI - Intravenous interferon-beta in chronic hepatitis B: promises. PMID- 8424417 TI - The p53 tumor suppressor gene as a common cellular target in human carcinogenesis. AB - The p53 gene is a 16-20 kb of cellular DNA located on the short arm of human chromosome 17 at position 17p13.1. This gene encodes a 375-amino acid nuclear phosphoprotein which involves in the regulation of cell proliferation. The p53 gene was originally regarded as a dominant oncogene because its overexpression resulted in the immortalization of rodent cells, and the p53 gene could transform rat embryonic fibroblasts in concert with an activated ras gene. It soon became clear, however, that many of the p53 clones that had been studied were in fact mutated versions of the gene, and the wild-type p53 actually acts as a tumor suppressor. Loss of normal p53 function has been associated with the cell transformation in vitro and the development of neoplasms in vivo. More than one half of human malignancies derived from the epithelial, mesenchymal, hematopoietic, and lymphoid tissues, as well as the central nervous system, analyzed thus far, were shown to contain an altered p53 gene. Most p53 gene alterations are the missense mutations, giving rise to an altered protein. These mutations are most frequently located in the evolutionally conserved areas. Furthermore, it has been demonstrated that the SV40 large T antigen, the adenovirus E1B protein, and papillomavirus E6 protein can bind to wild-type p53 protein and presumably lead to inactivation of this gene product as well. Therefore, the inactivation of normal (or wild-type) p53 is currently regarded as an important genetic pathway for human carcinogenesis generated by endogenous factors and exogenous carcinogens, as well as several tumor viruses. The current data on the p53 gene and its alterations in human malignancies, particularly those in the gastrointestinal tract, are reviewed. PMID- 8424418 TI - Chest pain associated with nutcracker esophagus: a preliminary study of the role of gastroesophageal reflux. AB - A review of our 402 motility records of patients undergoing evaluation of noncardiac chest pain identified 40 patients with the diagnosis of nutcracker esophagus. Gastroesophageal reflux was found in 13 of 20 patients (65%) who underwent pH studies, and endoscopy detected one patient with erosive esophagitis. Thus, at least 14 (35%) of our nutcracker esophagus patients had evidence of reflux. Twelve of these subjects agreed to enter an open-label therapeutic trial. After 8 wk of intensive antireflux treatment with high doses of ranitidine or omeprazole, repeat 24-h pH studies and endoscopy demonstrated normalization of pH parameters and healing of esophagitis in all patients. Ten (83%) patients obtained significant symptomatic improvement in frequency of pain episodes, number of days with pain, and pain severity. However, repeat manometry showed normalization of motor findings in only two (18%) patients. These observations warrant further placebo-controlled trials. Until more information is available, the results of this study suggest that gastroesophageal reflux should be excluded in patients with noncardiac chest pain and nutcracker esophagus before initiation of smooth muscle relaxant therapy. PMID- 8424419 TI - Erythromycin accelerates gastric emptying of indigestible solids and transpyloric migration of the tip of an enteral feeding tube in fasting and fed states. AB - Erythromycin has been shown to initiate gastric interdigestive migrating motor complexes, which are the motor events responsible for gastric emptying of indigestible solids. Hence, erythromycin should also accelerate gastric emptying of indigestible particles and facilitate transpyloric migration of the tip of an enteral feeding tube. Accordingly, we assessed the effect of erythromycin on these events, using a single-blind crossover study. Healthy subjects were nasally intubated with an enteral feeding tube. For fasting studies, the subjects remained fasted; in the fed studies, the subjects were fed a cheeseburger and fries after placement of the feeding tube. Then, ten 1-cm radiopaque plastic segments were swallowed by each subject, followed by an iv infusion of either erythromycin (200 mg over 20 min) or saline. Abdominal x-rays were then taken at regular intervals to document the location of the tube tip and the plastic segments. Erythromycin significantly shortened gastric emptying time of the indigestible particles during both fasting and fed states. Erythromycin also accelerated transpyloric migration of the tip of the feeding tube in both fasting and fed states. Hence, erythromycin can be beneficial when placement of a feeding tube in the small intestine is clinically desired. PMID- 8424420 TI - The effect of erythromycin in gastric emptying of solids and hypertonic liquids in healthy subjects. AB - We report our study of the effect of erythromycin on gastric emptying of solid and liquid meals in 10 healthy subjects. On different occasions, subjects consumed either a radiolabeled 50% glucose solution, or a radiolabeled standard solid meal after placebo, and after receiving 200 mg of erythromycin intravenously. Erythromycin accelerated the gastric emptying of the hypertonic liquid meal by significantly decreasing the duration of lag phase (p < 0.0001), by significantly increasing the emptying rate at the postlag period (p < 0.001), and by significantly decreasing the duration of the postlag period (p < 0.0001) and the meal remaining in the stomach at 15 (p < 0.05), 30 (p < 0.001), and 60 (p < 0.01) min postprandially. In addition, erythromycin administration induced a significant plasma fall at 15 (p < 0.05) and 30 (p < 0.01) min and a significant increase in pulse rate at 15 and 30 min (p < 0.01) after consumption of the hypertonic glucose solution, whereas three subjects experienced symptoms suggesting dumping syndrome. Furthermore, erythromycin administration enhanced the gastric emptying of solids by almost abolishing the duration of lag phase (p < 0.0001) and by reducing the overall t1/2 of emptying (p < 0.0001), whereas less food was retained in the stomach at 60 (p < 0.001) and 120 (p < 0.0001) min postprandially. Conversely, the postlag t1/2 of the solid meal emptying was not affected by erythromycin, as compared to placebo. We conclude that erythromycin has gastrokinetic properties, affecting the gastric emptying of both liquids and solids. PMID- 8424421 TI - The treatment of idiopathic and diabetic gastroparesis with acute intravenous and chronic oral erythromycin. AB - The objective of this study was to investigate the effects of intravenous erythromycin and chronic oral dosing of erythromycin on gastric emptying in patients with idiopathic or diabetic gastroparesis. Symptoms were assessed on oral dosing and during long-term follow-up in an ambulatory setting at a University referral center. Fourteen patients (10 idiopathic and four diabetic gastroparesis) were studied. Four patients left during the 4-wk study; two due to rash, one with cramps and vomiting on erythromycin, and one due to other medical problems. Ten patients completed the 4-wk study and commenced long-term therapy. Five of these patients experienced enough symptomatic relief to continue oral erythromycin long-term, being followed for an average period of 8.4 months. After initial documentation of delayed gastric emptying, patients received 6 mg/kg intravenous erythromycin lactobionate before a second gastric emptying study. Erythromycin base was then given orally at a dose of 500 mg tid-ac and qhs, with a final gastric emptying study performed after 4 wk. During long-term follow-up, erythromycin dosage was adjusted to minimize symptoms. Radionuclide-labeled gastric emptying of a solid meal was studied at baseline, following intravenous erythromycin, and after 4 wk of oral treatment with erythromycin. Symptom scores were assessed at baseline, at 4 wk, and then at 8-wk intervals. The percentage of the solid meal retained in the stomach at 2 h decreased from 85% +/- 11% (SD) at baseline to 20% +/- 29% following intravenous erythromycin (p < 0.001), and to 48% +/- 21% after 4 wk of oral therapy (p < 0.01 vs. baseline). There was a reduction in total symptom scores and a significant reduction in global assessment scores (p = 0.03). We conclude that erythromycin has a strong gastric prokinetic effect in both idiopathic and diabetic gastroparesis, and may represent a useful new therapeutic approach to this problem. PMID- 8424422 TI - Effect of intravenous erythromycin on postoperative ileus. AB - We attempted to determine whether the administration of erythromycin shortens the period of postoperative ileus by a prospective, double-blind, placebo-controlled study. Seventy-seven patients were randomized and included in the statistical calculations. The patients were stratified according to the operation performed (cholecystectomy, celiotomy, or other major abdominal operations). Forty-one patients (group 1) received 250 mg erythromycin intravenously every 8 h for nine doses upon admission to the recovery room. Thirty-six patients (group 2) received placebo. The time (in hours) to first passage of flatus, first liquid meal, first bowel movement, and total length of hospital stay was recorded. There was no significant difference between group 1 and group 2 in time to first flatus (54.9 +/- 29 vs. 53.9 +/- 27 h, respectively), first meal (70.4 +/- 44 vs. 71.7 +/- 65), first bowel movement (81.8 +/- 32 vs. 80.1 +/- 28), or length of hospital stay (185.2 +/- 183 vs. 182.1 +/- 163). Erythromycin, in the dosage tested in this study, does not seem to alter clinical parameters of gastrointestinal motility after an abdominal operation. New prokinetic agents may deserve further studies. PMID- 8424423 TI - A pilot study of long-term weekly interferon-beta administration for chronic hepatitis B. AB - Interferon-beta was given weekly for 24 wk, at a dose of 3 million units, intravenously, to 10 patients with chronic hepatitis B who were serologically positive for HBsAg and HBeAg. Their condition was followed for 6 months after the end of therapy. Both serum hepatitis B virus-associated DNA-polymerase activity and alanine aminotransferase level became significantly lower during therapy and during the 6 months after the end of therapy than at the beginning of therapy. In five of 10 patients, the seroconversion from HBeAg positive to anti-HBe positive had occurred by 6 months after the end of therapy, and in four of these five patients, serum alanine aminotransferase level became normal. Weekly interferon beta administration over 6 months seems effective in inducing seroconversion and in normalizing serum alanine aminotransferase level. PMID- 8424424 TI - Ranitidine has no effect on postbreakfast ethanol absorption. AB - The effect of ranitidine, the H2-receptor antagonist, on the systemic bioavailability of ethanol (0.3 g/kg body weight) taken orally 1 h after breakfast, was investigated in a randomized, placebo-controlled, double-blind cross-over study. Twenty normal male subjects (age, 19-26 yr) were studied on the morning of the 8th day of twice-daily dosing with either 150 mg ranitidine or placebo. Plasma ethanol concentration was measured by the alcohol dehydrogenase method from 0 to 240 min after oral ingestion of ethanol (100% ethanol made up to 200 ml orange juice). Compared with placebo, dosing with ranitidine resulted in nonsignificant changes in either the mean integrated 4-h plasma ethanol concentration (27.8 vs. 32.4 mg.h/dl), the peak plasma ethanol concentration (18.0 vs. 21.1 mg/dl), or the time to peak (43 vs. 40 min). There is no clinically important interaction between ranitidine and a low dose of ethanol taken orally 1 h after breakfast. PMID- 8424425 TI - A 4-week, multicenter, duodenal ulcer healing trial comparing four escalating doses of ranitidine. AB - In a randomized double-blind 4-wk trial, ranitidine doses of 300 mg at bedtime (hs), twice daily (bid), three times daily (tid), and four times daily (qid) were compared in 629 patients with endoscopically confirmed duodenal ulcer(s). Endoscopies were performed at baseline and after 4 wk of therapy. Per protocol analysis revealed wk 4 healing rates that were significantly increased (p < or = 0.001) for the bid, tid, and qid groups, compared with the hs group. All treatments provided early symptomatic (ulcer pain) relief. No significant differences in adverse events or laboratory abnormalities were observed between groups. Ranitidine 300 mg bid provides an alternative therapeutic approach for patient populations at risk for ulcer complications. These patients include those with the following: a past history of an upper gastrointestinal hemorrhage, perforation, obstruction, penetration, or giant (> 2.0 cm) duodenal ulcer. The elderly and those with chronic unresponsive ulcerations may also be included in this population. PMID- 8424426 TI - Enteral versus parenteral nutrition as adjunct therapy in acute ulcerative colitis. AB - To ascertain the role of total enteral nutrition, compared with total parenteral nutrition, as adjunct therapy to steroids in patients with severe acute ulcerative colitis, a prospective randomized trial was conducted in 42 of such patients. Inclusion criteria were the persistence of a moderate or severe attack of the disease (Truelove's index) after 48 h on full steroid treatment (prednisone 1 mg/kg/day). Patients were randomized to receive polymeric total enteral nutrition or isocaloric, isonitrogenous total parenteral nutrition as the sole nutritional support. Remission rate and need for colectomy were similar in both groups. No significant changes in anthropometric parameters were observed in either nutritional group at the end of the study. Median increase in serum albumin was 16.7% (-0.5% to +30.4%) in the enteral feeding group, and only 4.6% ( 12.0% to +13.7%) in the parenteral nutrition patients (p = 0.019). Adverse effects related to artificial nutritional support were less frequent (9% vs. 35%, p = 0.046) and milder in enterally fed patients. Postoperative infections occurred more often with parenteral nutrition (p = 0.028). These results suggest that total enteral nutrition is safe and nutritionally effective in severe attacks of ulcerative colitis. It is also cheaper and associated with fewer complications than parenteral nutrition. Total enteral nutrition should be regarded as the most suitable type of nutritional support in these patients. PMID- 8424427 TI - Lymphoblastoid alpha-interferon for chronic hepatitis C: a randomized controlled study. AB - Forty patients with chronic hepatitis C (CHC) were included in an open randomized controlled trial of lymphoblastoid alpha-interferon (L-IFN) versus no treatment. Twenty patients entered each group, and features of therapy and control cases were similar. L-IFN was given in low doses (1.5-4.5 megaunits) for 1 yr. In 16 of 20 patients treated with L-IFN (80%), but in only one of 20 nontreated cases (0.5%; p < 0.001), amino-transferase activities became normal. In four patients there was a reactivation of the disease during treatment after 4, 5, 6, and 8 months with normal aminotransferase levels. A posttherapy reactivation of hepatitis was observed in four additional cases after 1, 1, 1, and 3 months of follow-up. The other eight patients (40%) continued with normal aminotransferase levels for 1.52 +/- 0.74 (range, 1-2.1) yr after IFN doses were discontinued. In all treated patients except two nonresponders, but in only one of 20 nontreated cases (p < 0.001), Knodell's histological activity index decreased. Procollagen type III aminoterminal peptide levels did not change significantly in nontreated and nonresponder patients, diminished slightly in patients with a transient response, and normalized in cases with a long-standing response, suggesting that this serum test may be a reliable marker for monitoring response to IFN therapy in patients with CHC. Finally, L-IFN treatment induced significant increments in CD4/CD8 index, phytohemagglutinin-induced blastogenesis, and natural killer activity. This study shows that L-IFN diminish inflammatory and fibrogenic activity in most patients with CHC. In 40% of patients treated in this trial, a long-standing remission of the disease was observed. PMID- 8424428 TI - Natural course of chronic hepatitis C. AB - We studied 333 chronic hepatitis C patients to evaluate the natural course of this disease. Among 57 patients undergoing serial biopsies, 20 had chronic persistent hepatitis (CPH) at the first biopsy, and 10 of them progressed to chronic active hepatitis (CAH) or liver cirrhosis (LC) after 11 yr. Sixteen patients had CAH 2A, and this progressed to CAH 2B or LC in 10 cases over 9 yr. Among the 21 patients with CAH 2B, progression to LC was noted in 15 after 7 yr. Among the 100 patients observed for over 5 yr, the normalization of liver function for at least 3 yr was seen in only four patients. In two of these four patients, serum HCV-RNA was tested serially. Despite the sustained normalization of alanine aminotransferase levels, HCV-RNA continued to be detectable in one patient. We conclude that many patients with chronic hepatitis C eventually show progression of their disease after a long and symptomless course. PMID- 8424429 TI - Development of thyroid autoimmunity after administration of recombinant human interferon-alpha 2b for chronic viral hepatitis. AB - To investigate the frequency and clinical characteristics of autoantibody formation and development of autoimmune thyroid disease after interferon therapy, we measured the autoantibodies to thyrotropin receptor (TBII), thyroglobulin (ATA), and microsomal antigen (AMA) in 28 patients with histologically proven chronic viral hepatitis [25 males, three females; mean age 38.7 +/- 8.7 (SD) yr] receiving recombinant interferon-alpha 2b (IFN-alpha) treatment. Twenty patients with chronic hepatitis B (positive for HBsAg, HBeAg, and HBV DNA) and eight patients with chronic hepatitis C (positive for anti-HCV and HCV RNA) received IFN-alpha, 3 million units subcutaneously, three times a week for 6 months. Before, during, and up to 6 months after IFN-alpha therapy, thyroid hormone levels and titers of AMA, ATA, and TBII were measured every 2 months. None of them had thyroid dysfunction or antithyroid autoantibodies before IFN-alpha treatment. A 34-yr-old male patient developed Graves' disease during the last month of therapy. He required long-term antithyroid medications, even after discontinuation of IFN-alpha. Another 44-yr-old female patient developed AMA during IFN-alpha treatment; however, thyroid function remained normal and goiter did not develop in this patient. No other patient developed thyroid autoantibodies and thyroid dysfunction. In summary, only a small minority of patients will develop thyroid autoimmunity during IFN-alpha therapy, and much less often with this low dose of IFN-alpha. PMID- 8424430 TI - Biopsy study of polyps in the duodenal bulb. AB - To clarify the clinical and histological features of polyps in the duodenal bulb (DB), we studied, clinicopathologically, a total of 263 patients (179 male, 84 female) with polyps in the DB. The patients were 13-86 yr of age (average age, 57.1 yr). On endoscopy, a semipedunculated or pedunculated polyp with a relatively large size was seen in 33 patients, and small polyps, either single or multiple, were incidentally discovered in 230 patients. Histological examination of the biopsied specimens from polyps revealed gastric tissue in 182 cases (69.2%), cyst formation in 51 (19.4%), and other miscellaneous lesions (tubular adenoma, Brunner's gland hyperplasia, etc.) in 30 (11.4%). With regard to the gastric tissue, a "full-thickness" or "incomplete" body-type mucosa was identified in 27 cases of polyps, and hyperplastic surface epithelium was recognized in 155. Forty cases of hyperplastic surface epithelium-type polyps were subclassified as "pyloric"-type mucosa, because pyloric gland-like components seen under the surface epithelium were identical to those of the gastric antrum. It was reconfirmed that small polyps with little or no clinical significance could be produced by various histological features. PMID- 8424431 TI - Adenosine deaminase isoenzymes in liver disease. AB - To clarify the clinical significance of increased serum adenosine deaminase (ADA) activity, and its mechanisms in various liver diseases, ADA isoenzyme activities (ADA1 and ADA2) in serum and the peripheral blood mononuclear cells were studied. High serum ADA activities were found in patients with acute hepatitis, alcoholic hepatic fibrosis, chronic active hepatitis, liver cirrhosis, and hepatoma. The ADA2:ADA ratio was decreased in acute hepatitis, but was increased in chronic active hepatitis and liver cirrhosis. Clinically, ADA2 activity was correlated with serum gamma-globulin levels. In chronic active hepatitis, total ADA activities in the peripheral blood mononuclear cells were similar to those in controls. Furthermore, ADA2 activities after phytohemagglutinin (PHA) stimulation were significantly lower than those without PHA stimulation, although total ADA activities were increased after PHA stimulation. These findings suggest that serum ADA isoenzyme activities may be a new marker for liver disease, and that the increased serum ADA2 in chronic active hepatitis is unlikely to be the result of an increase in ADA2 production by activated peripheral blood mononuclear cells. PMID- 8424432 TI - Gross and microscopic findings in the human esophagus after esophageal variceal band ligation: a postmortem analysis. AB - Endoscopic sclerotherapy has been the therapy of choice for controlling acute variceal hemorrhage. Recently, endoscopic band ligation has been utilized for the management of esophageal varices with fewer complications and greater efficacy. We report the pathologic findings in the human esophagus 5 days after esophageal variceal band ligation. PMID- 8424433 TI - Stricture related to an inlet patch of the esophagus. AB - A 64-yr-old female was referred for evaluation of dysphagia. An esophagram showed a stricture in the proximal esophagus. On esophagogastroduodenoscopy, the patient was found to have a 3-cm stricture beginning at 14 cm from the incisors. This was initially dilated to 45-French with the Savary-Gilliard dilators. Biopsies and cytology of the stricture showed benign columnar epithelium. Subsequent dilations over the next 2 months resulted in dilation to 60-French with the esophageal dilators. Ulcerations related to this strictured area were healed over this period, with a sucralfate slurry, and the area of abnormality after healing appeared to be that of an esophageal inlet patch. PMID- 8424434 TI - Hepatitis A virus anicteric encephalitis coexistent with hepatitis C virus infection. AB - A 34-yr-old male presented with acute encephalitis. The encephalitis was due to an anicteric hepatitis A virus infection superimposed upon coexistent hepatitis C virus infection. Neurologic syndromes due to hepatitis A virus infection have all been associated with jaundice. Neurologic complications of hepatitis C virus infections have not been described. Identification of hepatitis A virus as an etiology of viral encephalitis can reduce the need for extensive costly evaluations, and unnecessary empiric antibiotics. Further clinical experience will determine if hepatitis C virus produces neurologic manifestations. PMID- 8424435 TI - Adenocarcinoma in childhood Barrett's esophagus: case documentation and the need for surveillance in children. AB - A 17-yr-old boy underwent esophagectomy for multifocal high-grade dysplasia and adenocarcinoma complicating Barrett's esophagus (BE). He is believed to be the first child or young adult to have prolonged healthy survival following resection of esophageal adenocarcinoma. Dysplasia in a short retained segment of his Barrett's mucosa appears to have regressed with acid-suppressing therapy. Of nine other reported cases of adenocarcinoma in young people 11-25 yr of age, all died. All had progressive dysphagia and an esophageal mass at presentation, unlike our patient who had only histologic evidence of cancer at presentation. This was found only after repeated and extensive biopsy of the esophagus. We conclude that adenocarcinoma does occur under age 25 yr as a complication of BE arising in childhood, and it may be curable if diagnosed early. Endoscopic surveillance with multiple stepwise biopsies, beginning at age 10 yr, is suggested in those few children who have BE with specialized mucosa and goblet cells. PMID- 8424436 TI - Unusual presentation of mucosal hypersensitivity secondary to gastroesophageal reflux disease. AB - We report an unusual presentation of gastroesophageal reflux disease in a 14-yr old boy with cervical dysphagia and vomiting immediately after swallowing. Reflux disease was diagnosed by the combination of eosinophils on esophageal biopsies and abnormal 24-h pH results. The cervical site of dysphagia demonstrated acid induced hypersensitivity to esophageal distension with water or air. The patient's symptoms resolved with marked acid suppression, which was made difficult because intact capsules of omeprazole initially could not be ingested. PMID- 8424437 TI - Eating disorders in inflammatory bowel disease. AB - Before or after their diagnosis, each of three patients with inflammatory bowel disease had an established eating disorder. Two had bulimia and one was presumed to have had anorexia/bulimia. In addition to the usual modes of weight control, such as vomiting and fasting, two of the patients, all of whom were lactose intolerant, used milk ingestion as a purgative. PMID- 8424438 TI - Giant gastric ulcers: an unusual manifestation of Crohn's disease. AB - We report a patient with Crohn's disease manifesting as recurrent giant gastric ulcers, with subsequent perforation and gastric outlet obstruction. The ulcers contained granulomas, and the patient was achlorhydric. To our knowledge, this is the first report of giant ulcers in a patient with gastric Crohn's disease. This case demonstrates another gastric manifestation of Crohn's disease, it documents nonmalignant gastric ulcers in the setting of achlorhydria, and it raises the possibility of Crohn's disease in the differential diagnosis of giant gastric ulcers refractory to medical management. PMID- 8424439 TI - Occult gastrointestinal bleeding in renal cell carcinoma: value of endoscopic evaluation. PMID- 8424440 TI - A case of intussusception and lymphoid hyperplasia in a patient with AIDS. AB - Idiopathic intussusception in adults is rare. In tropical climates, enteric infection is causally implicated. Three cases of intussusception in AIDS patients have been reported, two of which were associated with enteric infection. We report the fourth case of ileocolonic intussusception in an AIDS patient in whom lymphoid hyperplasia of the terminal ileum was found but no infection documented. The relationship between lymphoid hyperplasia and intussusception is discussed. The previous cases of AIDS and intussusception are reviewed. Idiopathic intussusception may become more prevalent as the number of AIDS cases increases, and must be considered in the differential diagnosis of abdominal pain in AIDS patients. PMID- 8424441 TI - The endoscopic diagnosis of iliac colonic fistula and review of literature. AB - Although the efficacy of upper gastrointestinal endoscopy in the diagnosis of a graft-enteric fistula is defined, the clinical utility of colonoscopy in this problem is not well described. We report a case of a 65-yr-old white male with gastrointestinal bleeding and a suspected graft-enteric fistula which was diagnosed by an immediate preoperative colonoscopic examination. The colonoscopy performed under general anesthesia in the operating room not only documented the presence and location of the graft colonic fistula but also illustrated the necessity for a more radical surgical resection before any planned revascularization. The current literature on the various methods of diagnosis and management of graft-enteric fistula was reviewed. PMID- 8424442 TI - A conservative approach to granular cell tumors of the esophagus: four case reports and literature review. AB - We present here four patients with six granular cell tumors of the esophagus: all are women, 22, 38, 47, and 49 yr old. In three of them, single polyps were encountered. In the fourth, age 38, two lesions were found in 1982, and a new one, a satellite polyp, has appeared after 5 yr of follow-up. All of the patients' conditions were followed up [mean time 65 months (16 months to 10 yr)]. In connection with these case reports, we include an up-to-date review of the world literature on this subject. PMID- 8424443 TI - Solitary rectal ulcer: is there a presymptomatic stage? PMID- 8424444 TI - Flow cytometry in Barrett's esophagus: when all is said and done, more is said than done! PMID- 8424445 TI - Hepatic failure after biliopancreatic diversion. PMID- 8424446 TI - Pathogenesis of chronic pancreatitis. PMID- 8424447 TI - Metronidazole-resistant Helicobacter pylori in a developing country. PMID- 8424448 TI - Ventricular expression of atrial and brain natriuretic peptides in dilated cardiomyopathy. An immunohistocytochemical study of the endomyocardial biopsy specimens using specific monoclonal antibodies. AB - Although brain natriuretic peptide is expressed in ventricles of failing hearts including dilated cardiomyopathy, its morphological localization is still unclear. In this study, we analyzed the immunohistocytochemical localization of atrial and brain natriuretic peptides in ventricles of dilated cardiomyopathy at both light and electron microscopic levels. Ventricular specimens were obtained by endomyocardial biopsy in 31 patients (26 with dilated cardiomyopathy and 5 controls without any specific cardiac disease). By light microscopic immunohistochemistry using specific monoclonal antibodies, all (26 of 26) of the left ventricular endomyocardial biopsy specimens and 31% (8 of 26) of the right ventricular specimens showed immunoreactivity for both of these natriuretic peptides in dilated cardiomyopathy. In contrast, none of the normal controls showed immunoreactivity for either of these peptides. The percentage of atrial natriuretic peptide-containing or brain natriuretic peptide-containing myocytes in the left ventricular specimens showed an inverse correlation with the left ventricular ejection fraction (r = -0.72 and r = -0.69, respectively). By electron microscopy, we identified specific secretory granules in ventricular myocytes from patients with dilated cardiomyopathy, but not in those from normal controls. Double immunocytochemistry using a two-face immunogold staining method revealed brain natriuretic peptide colocalized with atrial natriuretic peptide in the same ventricular granules. These findings suggest that brain natriuretic peptide is expressed in ventricular myocytes in response to hemodynamic stress in dilated cardiomyopathy. Brain natriuretic peptide may be, at least in part, synthesized simultaneously and secreted together with atrial natriuretic peptide by granules from failing ventricles, although the secretory turnover is different between these two peptides. PMID- 8424449 TI - Eosinophils are the major source of transforming growth factor-beta 1 in nodular sclerosing Hodgkin's disease. AB - Transforming growth factor-beta 1 (TGF-beta 1) is a multifunctional cytokine which promotes fibroblast growth and collagen synthesis, but suppresses growth and differentiation of immune lymphocytes and killer cells. Immunohistochemical detection of TGF-beta 1 in Hodgkin's disease (HD) has been shown to correlate with the histologic feature of nodular sclerosis, which is associated with a favorable prognosis (American Journal of Pathology 1990, 136:1209). In that study, TGF-beta 1 was localized mainly at the margins of broad collagen bands (presumably sites of new collagen synthesis) and in areas containing numerous Hodgkin/Reed-Sternberg cells (H/RS). In these areas, TGF-beta 1 protein was found on the membrane and occasionally within the cytoplasm of H/RS cells. To determine whether TGF-beta 1 is synthesized by H/RS cells or secondarily bound to their membrane and sometimes internalized, we performed in situ hybridization (ISH) using 1.5 Kb 35S-labeled anti-sense and sense RNA probes to TGF-beta 1. Paraffin embedded tissues of 10 cases from all histologic types of HD were examined. Somewhat unexpectedly, the major site of TGF-beta 1 mRNA was in eosinophils; TGF beta 1 mRNA was not detected in H/RS cells. TGF-beta 1 mRNA was found in eosinophils in all cases of nodular sclerosis but not in other types of HD, despite the presence of numerous eosinophils in mixed cellularity cases. The presence of TGF-beta 1 mRNA coincided with immunohistochemical detection of TGF beta 1 protein using antibody CC (1-30). These results confirm the role of TGF beta 1 in the histogenesis of nodular sclerosing HD and indicate that eosinophils are the major source of TGF-beta 1 in this type of HD. PMID- 8424450 TI - Neutrophil-mediated damage to human vascular endothelium. Role of cytokine activation. AB - Cytokine activation of cultured human vascular endothelial cells renders them hyperadhesive for blood leukocytes. Co-incubation of freshly isolated, unstimulated human blood neutrophils with confluent cytokine-activated human endothelial monolayers for 90 minutes results in extensive endothelial detachment and destruction of monolayer integrity. In contrast, unactivated endothelial monolayers remain intact. Using this in vitro model, we have explored the neutrophil-effector mechanisms involved in this injury. Coincubation in the presence of a serine protease inhibitor (phenylmethylsulfonyl fluoride) or specific elastase inhibitors (Ala-Ala-Pro-Val-chloromethyl ketone or alpha-1 protease inhibitor) markedly diminished injury. In contrast, scavengers or inhibitors of oxygen-derived free radicals (superoxide dismutase, catalase, mannitol, or sodium azide) were not protective. Purified human neutrophil elastase mimicked the effect of the neutrophils suggesting a key role for elastase in the neutrophil-mediated injury in this model. Interfering with direct neutrophil-endothelial cell contact by interposing a microporous barrier insert prevented endothelial cell detachment. Furthermore, this neutrophil-mediated detachment could be inhibited with interleukin-8, an action correlated with a decrease in neutrophil adhesion to activated endothelial monolayers. By defining the role of endothelial activation in neutrophil-mediated injury, this in vitro model may provide useful insights into potential therapeutic interventions designed to prevent disruption of the endothelial barrier function. PMID- 8424451 TI - Structural and biochemical changes in lungs of 3-methylindole-treated rats. AB - Effects of a single dose of 3-methylindole (3-MI) (250 mg/kg intraperitoneally) were studied at different times ranging from 12 hours to 2 weeks post-treatment (PT). Microscopic study revealed mild Clara cell injury 24 hours PT and mucus hyperplasia 24 hours to 2 weeks PT. Diffuse type I alveolar epithelial cell necrosis occurred at 48 hours, followed by type II cell hyperplasia. Septal edema and accumulation of interstitial and capillary polymorphonuclear leukocytes and perivascular mixed mononuclear inflammatory cells accompanied the injury and repair. A gradual resolution of lesions with persistent mononuclear inflammatory cellular clusters at septal junctions, focal septal fibrosis, and accumulation of alveolar macrophages was evident at 1 and 2 weeks PT. Collagen, measured as hydroxyproline, in 3-MI-treated rats was significantly increased to 130% and 139% of control (3.0 mg/lung) at 1 and 2 weeks PT, respectively. Biphasic peaks of plasma 6-keto-prostaglandin F1 alpha occurred at 12 to 24 hours and at 96 hours PT with 3-MI and thromboxane B2 was elevated 12, 48, and 96 hours PT. Right ventricular/left ventricular and septal weight was increased to 120% and 140% of the control 1 and 2 weeks PT. We concluded that 3-MI induces alveolar septal injury in the rat with relatively complete repair of the alveolar epithelium and residual mild focal septal fibrosis and pulmonary hypertension 2 weeks PT. Arachidonic acid-derived mediators and inflammation are associated with 3-MI induced lung injury. PMID- 8424452 TI - Direct and endothelial cell-mediated effect of cyclosporin A on the proliferation of rat smooth muscle cells in vitro. AB - Cyclosporin A (CsA) has been suggested to potentiate graft vascular disease by stimulation of smooth muscle cell (SMC) proliferation. Both the in vitro and in vivo data are discordant, showing both stimulatory and inhibitory effects of CsA on vascular SMC proliferation. The direct and endothelial cell-mediated effects of CsA on vascular SMC proliferation were examined in vitro using the incorporation of [3H]thymidine. All experiments were done in serum-free conditions. The exposure of SMC to CsA (0.0001 to 0.1 micrograms/ml) had no effect on proliferation. High doses of CsA (0.5 to 10.0 micrograms/ml) were toxic to the SMC and endothelial cells; 90% of SMC population died within 3 to 6 days of exposure to 10.0 micrograms/ml CsA. In the studies on the endothelial cell mediated effect of CsA, the endothelial cell-conditioned medium (ECCM) significantly increased SMC proliferation. This stimulatory effect was significantly attenuated when the ECCM was obtained from endothelial cells exposed to CsA. Endothelin (ET) is suggested to be an endothelial-cell-derived growth factor for SMC, and implicated as a possible cause of the uncontrolled proliferation of SMC during development of graft vascular disease. Exposure of SMC to levels of recombinant ET similar to the levels found in the ECCM (0.19 + 0.01 pg/ml) significantly increased SMC proliferation. CsA increased fivefold ET concentration in the ECCM. However, despite this rise in ET levels, there was a 45% decrease in SMC proliferation. In conclusion, CsA does not exert a direct modulatory effect on SMC proliferation in vitro, but may inhibit SMC proliferation indirectly via endothelial cell-derived factors. These unidentified factor(s) inhibit SMC proliferation and abolish the mitogenic effect of ET on SMC. PMID- 8424453 TI - Microinjection of synthetic amyloid beta-protein in monkey cerebral cortex fails to produce acute neurotoxicity. AB - The cerebral deposition of amyloid beta protein (A beta P) is an early pathogenetic event in Alzheimer's disease (AD). Recent studies suggest both neurotoxic and neurotrophic effects of A beta P in vitro. Because progressive A beta P deposition and surrounding neuritic dystrophy occur spontaneously in primates, we evaluated the in vivo effects of synthetic A beta P in monkey cortex. Experimental and control (reverse or substituted) peptides were stereotactically injected into multiple neocortical sites of adult rhesus monkeys in a vehicle of either artificial cerebrospinal fluid or acetonitrile. After 2 weeks, all injection sites were identified and characterized. A beta P antibodies specifically detected the injected A beta P1-40 peptide. Serial sections stained with silver and antineurofilament protein demonstrated comparable degrees of degenerating neurons, dystrophic neurites, and axonal spheroids associated with both experimental and control peptide injections. Alz 50 staining was sparse or absent in all sites. Similar results were obtained in an animal killed 3 months after injection. We conclude that specific cellular changes closely resembling the pathology of Alzheimer's disease were not detected in these acute experiments, and that control and experimental A beta P peptides produced indistinguishable effects. PMID- 8424454 TI - Effects of all-trans retinoic acid and Ca++ on human skin in organ culture. AB - In this study, we have established an organ culture model of human skin and examined the effects of both all-trans retinoic acid (RA) and extracellular Ca++ on the epidermal and dermal components of the organ-cultured skin. Our data show that while organ cultures maintained in serum-free, growth factor-free culture medium containing 0.15 mM Ca++ degenerated rapidly, those treated with concentrations of RA that have been shown previously to stimulate fibroblast and keratinocyte proliferation in monolayer culture (J Invest Dermatol 1989, 93:449; 1990, 94:717; Am J Pathol 1990, 136:1275) demonstrated a healthy appearance for up to 12 days. Degeneration of the control cultures was characterized by separation of the epidermis from the underlying dermis, progressive cell necrosis leading to a complete absence of viable cells from both the dermal and epidermal compartments, disintegration and fibrillation of the dermal connective tissue, and a cessation of protein synthesis. RA-treated organ cultures contained large numbers of healthy-appearing cells in both the epidermal and dermal compartments. One or several layers of viable basal cells in the epidermis could be seen at least through day 12. However, the upper layers of the epidermis frequently separated from the cells in the basal layer. The dermal connective tissue was histologically well-preserved. Furthermore, the level of protein synthesis was higher in the RA-treated cultures than in the control cultures. In addition to treating organ cultures with RA, other cultures were exposed to serum-free, growth factor-free culture medium containing 1.4 mM Ca++. The presence of the elevated Ca++ concentration also preserved cellular and connective tissue structures in the dermal and epidermal compartments. In comparison to RA there was better preservation of the overall epidermal structure. The upper layers of epidermal cells did not separate from the basal cells, and the various stages of epithelial differentiation could be seen. Histologically, the dermis was well preserved in the presence of elevated extracellular Ca++. Specimens treated with a combination of Ca++ and RA demonstrated features consistent with the features induced by each treatment separately. This included an expanded basal layer of epithelial cells and a prominent keratotic layer with a fairly orderly pattern of differentiation. The tendency of the upper epidermis to separate from the basal cells was partially mitigated. Taken together, these data indicate that both RA and extracellular Ca++ act to prevent the degeneration of human skin in organ culture but probably do so through different mechanisms. PMID- 8424455 TI - Role of glomerular epithelial cell injury in the pathogenesis of glomerular scarring in the rat remnant kidney model. AB - We investigated the roles of glomerular epithelial cell (GEC) pathology and dysfunction in the pathogenesis of glomerular scarring and attempted to separate them from direct hypertensive injury in the 5/6 nephrectomy (RK) model of glomerular injury. Male WKY rats weighing 200 g were studied 6 weeks after RK, when approximately one-half had developed systemic hypertension (systolic blood pressure > or = 150 mm Hg) (HT), and one-half were normotensive (NT). The incidence of glomerular necrosis and scarring was greatest in the HT rats (P = 0.0259), and vascular necrosis was only seen in 4 of 11 HT rats. The RK group had increased glomerular diameters (HT, 174 mu mean; NT, 171 mu; sham, 142 mu; P = 0.0014 by analysis of variance). There was foot process effacement in the HT and NT groups (HT, 104 filtration slits/100 mu glomerular basement membrane; NT, 112 mu; sham, 143 mu; P < 0.005 by analysis of variance), but GEC separation from the glomerular basement membrane was not significant in either HT or NT rats. GEC function was determined from protamine-heparin aggregate disappearance curves, and the curves, representing GEC endocytosis, were not different in either HT or NT groups compared with the sham-operated groups. These findings suggest that GEC function is preserved in RK, and the changes in glomerular size and GEC morphology are nonlethal and adaptive. The morphological appearance of the acute glomerular and vascular lesions and their presence only in HT animals is consistent with a hypertensive pathogenesis. The glomerular sclerosis seen in both HT and NT may result from either resolution of acute lesions with scarring and/or adaptive changes in glomerular structure and cellular functions other than the GEC clearance function we studied. PMID- 8424456 TI - Cleavage of human and mouse cytoskeletal and sarcomeric proteins by human immunodeficiency virus type 1 protease. Actin, desmin, myosin, and tropomyosin. AB - HeLa cell actin was cleaved by human immunodeficiency virus type 1 protease when in its soluble, globular form (G-actin). No cleavage of the polymerized, filamentous form of actin (F-actin) was observed when examined by denaturing gel electrophoresis; however, electron microscopy revealed a low level of cleavage of F-actin. Immunoblotting of mouse skeletal and human pectoral muscle myofibrils treated in vitro with human immunodeficiency virus type 1 protease showed that myosin heavy chain, desmin, tropomyosin, and a fraction of the actin were all cleaved. Electron microscopy of these myofibrils demonstrated changes consistent with cleavage of these proteins: Z-lines were rapidly lost, the length of the A bands was shortened, and the thick filaments (myosin filaments) were often laterally frayed such that the structures disintegrated. Nonmuscle myosin heavy chains were also cleaved by this enzyme in vitro. These data demonstrate that this protease can cause alterations in muscle cell ultrastructure in vitro that may be of clinical relevance in infected individuals. PMID- 8424457 TI - Accumulation of surfactant protein D in human pulmonary alveolar proteinosis. AB - Surfactant protein D (SP-D) is a collagenous calcium-dependent carbohydrate binding protein that is structurally related to the serum mannose-binding proteins and pulmonary surfactant protein A. SP-D was initially characterized as a biosynthetic product of freshly isolated rat type II cells and first purified in chemical amounts from bronchoalveolar lavage of rats with silica-induced alveolar lipoproteinosis. The present studies describe the characterization of human SP-D isolated from therapeutic bronchoalveolar lavage of patients with pulmonary alveolar proteinosis. Human proteinosis SP-D was extracted from the 10,000 x g pellet of bronchoalveolar lavage with 100 mmol/L glucose or ethylenediamine tetraacetic acid, and specifically bound to and eluted from maltosyl-agarose. The protein cross-reacted with monospecific antibodies to rat SP-D by enzyme-linked immunosorbent assay and immunoblot and eluted near the position of rat SP-D on reverse-phase high performance liquid chromatography. When chromatographed on 4% agarose (A-15M) in the presence of ethylenediamine tetraacetic acid, the solubilized human proteinosis SP-D eluted near the void volume and earlier than rat SP-D dodecamers or human SP-D multimers in the lavage supernatant. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting of proteins in the lavage pellet with antibodies to the carbohydrate-binding domain of proteinosis human SP-D demonstrated covalently cross-linked multimers of SP-D monomers (43 kd, reduced) and multimers of trimeric components stabilized by disulfide and non-disulfide bonds. These studies describe the isolation and biochemical characterization of human SP-D and demonstrate the abnormal accumulation of this protein in the air spaces of patients with alveolar proteinosis. PMID- 8424458 TI - Detection of a neuron-specific 9.0-kb transcript which shares homology with antisense transcripts of HIV-1 gag gene in patients with and without HIV-1 infection. AB - The pathogenesis of the neurologic abnormalities associated with the acquired immune deficiency syndrome (AIDS) is poorly understood. Although human immunodeficiency virus type 1 (HIV-1) transcripts have been detected in endothelial cells and macrophages of the central nervous system in patients with AIDS, infection of neuronal cells by HIV-1 has not been established. The purpose of this study was to localize HIV-1 transcripts in the central nervous system. 3H and digoxigenin-UTP-labeled riboprobes generated from a 942-bp fragment of DNA from the 5' end of the HIV-1 gag sequence were used for in situ hybridization. The antisense riboprobe hybridized to lymphoid cells in the sections of kidney and spleen obtained from patients with AIDS, as well as to the HIV-1-infected A3.01 cell line. The control sense probe did not hybridize to these same cells. In contrast, no detectable hybridization was observed to neuronal cells when the antisense probe was applied to sections of brain obtained from patients with and without AIDS. To our surprise, however, specific hybridization was observed to neuronal cells when the control sense probe was applied. This hybridization with the control sense probe was seen in both patients with and without HIV-1 infection. Northern blot analysis confirmed the in situ hybridization results; a unique 9.0-kb transcript was detected exclusively in brain tissue. These data suggest that there is a neuron-specific 9.0-kb transcript that shares extensive homology with antisense gag HIV-1 sequences and that this transcript is expressed in neuronal cells of both HIV-1-infected and noninfected individuals. The biological significance of this 9.0-kb transcript is unknown, but it may play an important role in the interactions of HIV-1 with neuronal cells. PMID- 8424459 TI - Poly-L-lysine-gold probe for the detection of anionic sites in normal glomeruli and in idiopathic and experimentally-induced nephrosis. A comparative ultrastructural study. AB - Anionic sites play a key role in the charge selectivity of glomerular filtration as well as in the maintenance of the structural integrity of the visceral epithelium and podocytes. Alterations in these sites are believed to be a major factor underlying human idiopathic nephrosis and puromycin-nephrosis in the rat. The poly-L-lysine-gold complex was used for the ultrastructural detection of anionic sites in renal glomeruli of patients with idiopathic nephrosis as well as of rats with puromycin-induced nephrosis, allowing for study of the changes occurring in the anionic sites during nephrosis and for the comparison between human disease and this experimental model. In both normal human and rat controls, the probe was detected on epithelial and endothelial cell surfaces and on the glomerular basement membrane, mainly in both laminae rarae. In proteinuric rats, a decrease in labeling intensity was noted on podocyte membranes and in the lamina rara externa, with a corresponding increase in the more central areas of the glomerular basement membrane. These changes were not as evident in proteinuric humans. Furthermore, a reduction in labeling density was noted in the glomerular basement membrane of proteinuric animals, although this could not be substantiated in human tissues. Poly-L-lysine-gold is a useful probe for anionic sites in fixed tissues, and allows for comparison between human disease and its experimental counterpart. PMID- 8424460 TI - Fibroblast migration in fibrin gel matrices. AB - In healing wounds and many solid tumors, locally increased microvascular permeability results in extravasation of fibrinogen and its extravascular coagulation to form a fibrin gel, with concomitant covalent cross-linking of fibrin by factor XIIIa. Subsequently, inflammatory cells, fibroblasts, and endothelial cells migrate into the gel and organize it into granulation tissue and later into mature collagenous connective tissue. To gain insight into some of the cell migration events associated with these processes, we developed a quantitative in vitro assay that permits the study of fibroblast migration in fibrin gels. Early passage human or rat fibroblasts were allowed to attach to tissue culture dishes and then were overlaid with a thin layer of fibrinogen that was clotted with thrombin. Fibroblasts began to migrate upwards into the fibrin within 24 hours and their numbers and the distance migrated were quantified over several days. The extent of fibroblast migration was affected importantly by the nature of the fibrin clot. Fibroblasts migrated optimally into gels prepared from fibrinogen at concentrations of -3 mg/ml; ie, near normal plasma fibrinogen levels. Migration was greatly enhanced by extensive cross-linking of the fibrin alpha-chains by factor XIIIa, as occurs when clotting takes place in vivo. When fibrinogen was clotted in Dulbecco's modified Eagle's medium, gamma-chains were cross-linked, but alpha-chain cross-linking was strikingly inhibited, and fibroblasts migrated poorly. Gels prepared from factor XIII-depleted fibrinogen exhibited neither alpha-nor gamma-chain cross-linking and did not support fibroblast migration. Further purification of fibrinogen by anion exchange high pressure liquid chromatography depleted fibrinogen of fibronectin, plasminogen, and other impurities; this purified fibrinogen clotted to form fibrin gels that supported reproducible fibroblast migration. PMID- 8424461 TI - Immunoglobulin specificity of low grade B cell gastrointestinal lymphoma of mucosa-associated lymphoid tissue (MALT) type. AB - The specificity of the tumor cell immunoglobulin in three cases of low grade B cell gastrointestinal mucosa-associated lymphoid tissue (MALT) lymphoma has been studied. Using anti-idiotypic antibodies to detect the reactivity of tumor immunoglobulin in tissue sections from the patients and other individuals, we observed specificity for normal tissue components in all three cases studied. Reactivity in one case was with follicular dendritic cells, in the second case with a novel antigen on mucosal post capillary venules, and, in the third case, a broad pattern of reactivity was observed. This study suggests that autoimmunity may play a role in the pathogenesis of gastric lymphoma. PMID- 8424462 TI - Chromosomal abnormalities in leiomyosarcomas. AB - Thirty-eight tumors from 30 patients diagnosed as leiomyosarcoma were cytogenetically assessed after short term culture. The specimens were obtained from the retroperitoneum, gastrointestinal tract, and extremities. Chromosomal abnormalities were present in 18 tumors from 13 patients; 15 tumors had clonal changes, whereas 3 tumors had numerous nonclonal changes. Ten tumors from 10 patients had normal karyotypes and no results were obtained in 10 other tumors from 7 patients. Of the tumors with clonal chromosomal aberrations, 4 had near diploid (3 hypo- and one hyperdiploid) modes, 8 were polyploid, and 3 were bimodal. No specific karyotypic change appeared to characterize the leiomyosarcomas, although involvement of some chromosomes appeared more frequent than others. A comparison of our findings with those reported in the literature revealed certain consistent structural rearrangements involving chromosomes 1, 7, 10, 13, and 14 at bands 1p36, 1p32, 1p13, 1q32, 7p11.1-p21, 7q32, 10q22, 13q14, and 14p11, respectively. Other bands less frequently rearranged were 3p13-p22, 3q21, 4q13-q23, 6q15-q21, 7q11.2-q22, 12q13-q14, 17q12-q25, 19q13.3-q13.4, and 20q12-q13.1. Numerical changes included recurrent loss of chromosomes 4, 9, 14, 15, 16, 18, 21, and 22. Identification of the abnormalities of these chromosomes is important in that it may predict the existence of oncogenes, tumor suppressor genes, and/or growth factor genes at these sites. Subsequent molecular analysis might then lead to the identification of the genes involved and ultimately to a better understanding of the pathogenesis of leiomyosarcomas. PMID- 8424463 TI - Rabbits with elevated serum C-reactive protein exhibit diminished neutrophil infiltration and vascular permeability in C5a-induced alveolitis. AB - In previous studies, we have shown that C-reactive protein (CRP) inhibits chemotaxis of neutrophils to complement fragments in vitro. To evaluate the effect of CRP on C5a-induced inflammation in vivo, a rabbit model of alveolitis was used. Rabbits pretreated with subcutaneous injections of croton oil had serum CRP increase from undetectable levels to 270 +/- 70 micrograms/ml 48 hours later. Rabbits were intubated and C5a des arg (10 micrograms/ml) instilled directly into the lungs via an endotracheal tube. Four to six hours later, the animals were killed and bronchoalveolar lavage performed. Rabbits pretreated with croton oil had significantly (P < 0.01) reduced C5a des arg-stimulated neutrophil infiltration (30 +/- 5%) into alveolar air spaces compared to untreated rabbits (64 +/- 9%). Increased numbers of total leukocytes in the alveolar washes coincided with increased neutrophil numbers whereas alveolar macrophages remained unchanged in all groups. Rabbits pretreated with croton oil also had a significant decrease (P < 0.05) in total protein (320 +/- 50 micrograms/ml) in lavage fluid after C5a instillation compared with untreated animals (850 +/- 140 micrograms/ml). In vitro, rabbit CRP (50 micrograms/ml) added to normal rabbit serum significantly (P < 0.05) inhibited chemotaxis of human neutrophils by 41%. Finally, direct intravenous pretreatment of rabbits with purified CRP also significantly reduced C5a-induced alveolitis. The CRP-C5a group had 33 +/- 10% neutrophil infiltration, a significant (P < 0.01) reduction from the C5a group (71 +/- 6%). The total protein content of the CRP-C5a rabbits was 986 +/- 165 micrograms/ml in the lavage fluid, which was significantly (P < 0.05) lower than the C5a group (1645 +/- 363 micrograms/ml). Therefore, CRP inhibits the development of neutrophil alveolitis and protein leakage in vivo and inhibits neutrophil chemotaxis in vitro. These data indicate that CRP offers a protective effect in neutrophil-mediated lung injury by reducing neutrophil influx and protein leak. PMID- 8424464 TI - Centrocytic lymphoma: a morphometric study with comparison to other small cleaved follicular center cell lymphomas and genotypic correlates. AB - Centrocytic lymphoma (ML,CC) is distinguished from other cleaved follicular center cell (FCC) lymphomas in the Kiel classification by the lack of noncleaved FCC and not by morphological differences between the centrocytes (cleaved cells). Immunophenotypic and genotypic studies, however, have shown that the centrocytes in ML,CC are distinct from those of other small cleaved FCC lymphomas (ML,FCC,SC). To morphologically compare the cells of ML,CC with nine previously studied ML,FCC,SC and to relate the findings in ML,CC to the varied descriptions of lymphomas of intermediate differentiation, a morphometric analysis of 22 ML,CC was performed. Nuclei in ML,CC were, on average, significantly larger, rounder, and had less frequent nucleoli than those in ML,FCC,SC; however, the proportion of small round lymphocytes did not differ. Among the ML,CC, the only apparent immunophenotypic/genotypic correlate that was identified was greater nuclear ellipticity for the biopsies lacking chromosome 11q13 bcl-1 or PRAD1 rearrangement. Repeat biopsies in four patients with ML,CC showed an increase in nuclear size. These data demonstrate that a lack of transformed cells is not the only morphological difference between ML,CC and ML,FCC,SC. The morphological distinction, however, is not based on the proportion of small round lymphocytes present. In addition, the morphometric parameters illustrate the nuclear variability among ML,CC and demonstrate how the disease may evolve over time. PMID- 8424465 TI - Clear cell sarcoma of the kidney expresses insulinlike growth factor-II but not WT1 transcripts. AB - Two cases of clear cell sarcoma of the kidney (CCSK), five Wilms' tumors (WTs), and three fetal kidneys were studied by molecular hybridization to elucidate the histogenesis of CCSK. Northern blot and in situ hybridization demonstrated that all the CCSKs, WTs, and fetal kidneys contained abundant insulinlike growth factor-II (IGF-II) transcripts, whereas WTs and fetal kidneys--but not CCSKs- showed significant expression of WT1 gene, a candidate tumor suppressor gene implicated in the etiology of WTs. Comparative analysis of in situ hybridization of IGF-II and WT1 transcripts in CCSKs and fetal kidneys revealed that CCSK cells showed similar hybridization patterns to primitive metanephrogenic blastemal cells and early stromagenic cells. The data strongly suggest that CCSK is distinct from WT and may be derived from undifferentiated metanephrogenic blastemal cells with potential to differentiate into a stromal cell lineage. The result also suggests that this unique tumor should, more appropriately, be known as stromal--rather than clear--cell sarcoma of the kidney. PMID- 8424466 TI - Cellular localization of type 1 plasminogen activator inhibitor messenger RNA and protein in murine renal tissue. AB - Type 1 plasminogen activator inhibitor (PAI-1) may be markedly increased in the plasma of patients with endotoxemia and/or renal disease. To investigate renal PAI-1 production during acute endotoxemia, a murine model system was used. Mice were injected with either saline alone or saline containing 50 micrograms endotoxin, and sacrificed 3 hours later and their tissues analyzed for PAI-1 messenger RNA (mRNA) and antigen. Northern blot analysis confirmed that the level of renal PAI-1 mRNA was greatly increased in the endotoxemic mice relative to the saline controls. In situ hybridization was then performed to determine the cellular localization of PAI-1 mRNA within the renal tissues. In the control kidneys, low levels of PAI-1 mRNA were detected in the renal papilla and in the muscular walls of renal arteries. However, in the endotoxemic mice, an intense hybridization signal for PAI-1 mRNA was observed in glomerular and peritubular cells. These cells also stained positively for von Willebrand factor antigen, an endothelial cell-specific marker. The PAI-1 mRNA hybridization signal could further be observed in peritubular endothelial cells in the medulla and in endothelial cells of veins and arteries throughout the kidney. Immunochemical analysis revealed that PAI-1 antigen co-localized to the cytoplasm of cells expressing PAI-1 mRNA. This study provides the first direct evidence that PAI-1 is induced in endothelial cells of the kidney during endotoxemia in vivo and suggests a role for PAI-1 in the pathogenesis of renal disease. PMID- 8424467 TI - Involvement of transforming growth factor-beta in the formation of fibrotic lesions in carcinoid heart disease. AB - Carcinoid heart disease is a complication of a neuroendocrine carcinoid tumor. Morphologically, it is characterized by the formation of fibrotic plaques with deposition of extracellular matrix in the subendocardium, frequently causing heart valve dysfunction and cardiac failure. Because members of the transforming growth factor-beta (TGF-beta) family are known to stimulate fibroblasts in their production of extracellular matrix, we investigated the expression of the three isoforms of TGF-beta and the binding protein for latent TGF-beta 1 (LTBP) in carcinoid plaques of the right side of the heart, as well as from control tissue, using immunohistochemistry. Tissue specimens were obtained intraoperatively from nine consecutive patients undergoing valve replacement surgery. TGF-beta 1 and TGF-beta 3 were detected in the fibroblasts of all plaques analyzed, whereas TGF beta 2 was only rarely expressed. The localization of LTBP was partly concordant with that of TGF-beta 1, but the positive staining for LTBP was extracellular. Sections from unaffected heart tissue contained few fibroblasts in the subendocardium, showing only weak or no immunostaining for TGF-beta 1, -beta 2, and -beta 3 and no staining for LTBP. These results suggest that TGF-beta may play a role in the proliferation of fibroblasts and their matrix production in carcinoid heart lesions. PMID- 8424468 TI - Coordinate synthesis of stromelysin, interleukin-1, and oncogene proteins in experimental osteoarthritis. An immunohistochemical study. AB - Metalloproteases appear to play an important role in the pathophysiology of osteoarthritis (OA) and their expression is believed to be regulated by cytokines such as interleukin-1 (IL-1). Nuclear oncogene products are suggested as mediators through which IL-1 induces metalloprotease gene expression. Little data are available on the in vivo involvement of these agents in the pathophysiology of OA. This study examined by immunohistochemistry, using specific antibodies, the distribution of stromelysin, IL-1 alpha, IL-1 beta, and oncogene products (c FOS, c-JUN, and c-MYC) in synovium and cartilage from normal and experimental canine models of OA. In the OA synovium, stromelysin and IL-1 were localized in the cytoplasm of superficial synovial lining cells, infiltrating mononuclear cells, and endothelial and smooth muscle cells of the blood vessels, whereas oncoproteins were detected predominantly in the synovial lining cells. Normal synovial membranes demonstrated low levels of specific staining in synovial lining cells with occasional staining of blood vessel cells for IL-1 alpha, IL-1 beta, and stromelysin. In OA cartilage, chondrocytes at the superficial and middle layers as well as in fibrillated areas were found to be involved in the synthesis of stromelysin, IL-1, and oncoproteins. Diffuse staining of stromelysin and IL-1 beta in OA cartilage matrix was also identified. In normal cartilage, only a few chondrocytes at the superficial layer showed a low level of antigens. These results demonstrate the in vivo concomitant cellular and/or matrical presence of stromelysin, IL-1, and oncogene proteins in tissues from experimentally induced OA with the most intense staining at the sites of cartilage erosion and synovial proliferation. These findings suggest that they may be involved in the pathophysiology of OA, and that the regulatory mechanisms involved in the expression of these proteins may be associated. PMID- 8424469 TI - Early history of tuning-fork tests. PMID- 8424470 TI - Antidepressant treatment of tinnitus patients: report of a randomized clinical trial and clinical prediction of benefit. AB - Ninety-two middle-aged and elderly patients with disabling tinnitus participated in a double-blind randomized clinical trial comparing nortriptyline (a tricyclic antidepressant) to placebo. The study was stratified for presence (n = 38) or absence (n = 54) of current major depression (by DSM-III criteria). Both active drug and placebo were given for 6 weeks following a dose adjustment phase; the median nightly dose of nortriptyline was 100 mg. The two primary outcome variables were global satisfaction questions: "Has the medication helped you in any way?" and "Has your tinnitus improved?" Sixty-seven percent of nortriptyline patients stated the drug had helped them, versus 40 percent of placebo patients (chi-square = 7.14, p = 0.008). However, tinnitus severity was not significantly affected by nortriptyline (active: 43%; placebo: 30%; chi-square = 1.567, p = N/S). Benefit was more likely to be reported by depressed patients, by patients with insomnia, by women, and by patients without cervical musculoskeletal disease. Nortriptyline is useful in some patients with disabling tinnitus, but has not been shown to directly affect tinnitus sensation. Placebo effects were strongly significant and must be considered important in tinnitus therapy. It is difficult to specify the most appropriate outcome measures for tinnitus therapeutic trials. PMID- 8424471 TI - Betahistine-induced vascular effects in the rat cochlea. AB - Betahistine (BH) has been used widely to treat cochlear disorders, such as tinnitus and Meniere's disease. The mechanism of action of BH in the cochlea is assumed to be based on its histamine-like effect on H1 receptors in the cochlear vasculature, leading to an increased cochlear blood flow (CBF). Recently it has been shown that BH can strongly affect H3 heteroreceptors (a novel histamine receptor subclass) in the periphery, via an autonomic ligand. This mechanism may also contribute to the BH effects on CBF. This study was to validate BH effects in the cochlear vasculature and to investigate the possible mechanisms of action of this drug in the inner ear vasculature. We assessed the effects of BH on CBF with the laser Doppler flowmeter in 23 rats and concluded that BH affects vascular conductivity in the cochlea in a dose-dependent fashion; betahistine diffuses through the round window, but does not have access to vascular receptors or ligands once in the labyrinthine fluids; and the H1 receptors mediate the systemic and peripheral vascular effects of BH, whereas the cochlear effect involves cholinergic receptors. PMID- 8424472 TI - Third party utilization review: the corporate practice of medicine. PMID- 8424473 TI - Posterior internal auditory canal closure following the retrosigmoid approach to the cerebellopontine angle. AB - The retrosigmoid approach is utilized in a variety of cerebellopontine angle and internal auditory canal procedures. Drill curettage of the posterior internal auditory canal enhances lateral exposure, however, this step may also increase the patient's risk for postoperative cerebrospinal fluid (CSF) otorrhea. Obliteration of perilabyrinthine air cells is technically difficult and muscle graft displacement frequently occurs. A technique for posterior petrous dural flap stabilization of a temporalis muscle plug has proved successful in decreasing the risk of postoperative CSF fistula following retrosigmoid surgery. Temporal bone air-cell anatomy, as it relates to retrosigmoid, posterior internal auditory canal surgery is reviewed. Our technique for internal auditory canal closure, with bone wax, bone pate, muscle grafts, and petrous ridge dural flaps is outlined. PMID- 8424474 TI - Otoacoustic emissions in children with normal ears, middle ear dysfunction, and ventilating tubes. AB - The clinical utility of otoacoustic emissions (OAEs) has been well established in adults. The purpose of this investigation was to determine the efficacy of OAE testing in children. Distortion-product OAE (DPOAE) audiograms, response/growth functions, and transiently evoked OAEs elicited with clicks were measured from the ears of both healthy volunteers, aged 4 to 13 years, and children with confirmed middle ear disorders. These measures established the means and variabilities for DPOAE and noise-floor amplitudes of normal and diseased young ears. Compared with adult emissions, the healthy young ears exhibited greater mean DPOAE and noise-floor amplitudes. In contrast, ears with type B and type C tympanogram patterns showed absent or markedly reduced OAE amplitudes, when compared with emissions measured in their control counterparts. Finally, ears with ventilating tubes exhibited OAE amplitudes lower than amplitudes from healthy ears, but higher than those of the untreated diseased ears. Although these findings imply that using OAEs to test the outer hair-cell reserve of infected ears is problematic, emitted responses provide useful information concerning the normalcy of middle ear function. PMID- 8424475 TI - Ultrastructural organization of calcitonin gene-related peptide immunoreactive efferent axons and terminals in the vestibular periphery. AB - The ultrastructural distribution of calcitonin gene-related peptide immunoreactivity (CGRPi) was examined in sections of decalcified temporal bones in order to study the complex peptidergic innervation patterns of this efferent neuromodulator in the peripheral vestibular system of the rat. A new method of preembedding immunoelectron microscopy was developed to accomplish this study. Unmyelinated CGRPi axons, measuring 1 to 3 microns in diameter, passed among the primary afferent fibers in Scarpa's ganglion, and these fibers continued through the subepithelial regions of the vestibular end-organs. Within the neurosensory epithelia of the maculae and cristae, the CGRPi axons ramified to produce numerous CGRPi terminals. Immunoelectron microscopic localization of CGRPi terminals in the maculae and cristae revealed an extensive innervation pattern on the afferent vestibular pathway. Calcitonin gene-related peptide immunoreactive terminals made synaptic contacts with the unmyelinated portions of the primary afferent vestibular fibers innervating both type I and type II hair cells. Abundant synaptic contact between CGRPi terminals and the chalices surrounding type I hair cells was observed. Rare direct contact between CGRPi terminals and type I or type II hair cells was observed. In addition, vesiculated efferent terminals without CGRPi were seen contacting type II hair cells. These data suggest that the efferent vestibular system has a much more complex innervation pattern on the afferent vestibular pathway than previously believed. PMID- 8424476 TI - Intravenous fluorescein for detection of perilymphatic fistulas. AB - It has been reported, in animal models and recently in human beings, that intravenous fluorescein is taken up in perilymph and may be useful as a tracer for the detection of perilymphatic fistulas. We attempted to reproduce the results of these animal experiments. Twenty-one middle ears of eight cats and four dogs were exposed. Fluorescein was given intravenously. Fluorescence was initially noted in transudates pooling in the oval and round window niches. Fistulas created with a straight pick produced a bright fluorescence in the leaking fluids, possibly from a ruptured small membrane vessel. Fistulas created with the carbon dioxide laser and with complete hemostasis demonstrated no fluorescence. We concluded that intravenously administered fluorescein causes dramatic fluorescence of vessels and transudates that may be interpreted falsely as fluorescence of perilymph. Fluorescence was not evident in perilymph when complete hemostasis was obtained. PMID- 8424477 TI - Magnetic resonance angiography: analysis of vascular lesions of the temporal bone and skull base. AB - Contrast-enhanced, high resolution, computed tomography (CT), along with gadolinium-enhanced magnetic resonance imaging (MRI), provide the skull-base surgeon with essential information for diagnosis and surgical planning. Evaluation of skull-base vasculature has traditionally been performed with angiography. Four-vessel angiography is an invasive procedure that subjects the patient to the potential risks of vascular injury, stroke, renal damage, anaphylaxis, and radiation. The development of gradient-recall echo pulse sequence in MRI provides a means to evaluate the vasculature with a noninvasive, low-risk technique. We review the experience of the House Ear Clinic with magnetic resonance angiography (MRA) in evaluating cranial-base abnormalities including paragangliomas, aberrant vessels, and dural sinus thrombosis. Based on a review of 25 cases, we conclude that MRA is a useful procedure for evaluation of major arteries, veins, and dural sinuses. The technique has been less helpful in defining small vessel detail such as feeding blood supply and tumor staining. PMID- 8424478 TI - Tympanic and transtympanic electrocochleography in acoustic neuroma and vestibular nerve section surgery. AB - Eighth nerve action potential (AP) amplitudes and latencies and cochlear microphonic (CM) amplitudes were compared using tympanic and transtympanic electrocochleography (ECOG) in two patient groups. Tympanic ECOG was performed with a wick electrode placed on the tympanic membrane (TM). Transtympanic ECOG was performed with a needle electrode placed on the promontory of the anesthetized patient. Eighteen subjects were tested by tympanic ECOG as part of a preoperative assessment for either acoustic neuroma removal or transection of the vestibular portion of the eighth cranial nerve. Surgery occurred within 1 week of the preoperative evaluation. Intraoperative auditory monitoring was performed using transtympanic ECOG. Baseline recordings were compared to the preoperative tympanic ECOG data. Stimuli were condensation and rarefaction clicks and tone bursts, presented by an insert earphone. As expected, the two methods resulted in essentially identical response latencies and large amplitude differences, although the response amplitudes were extremely variable. The AP amplitude and the CM amplitude did not increase by the same factor with the transtympanic (TT) electrode compared to the tympanic electrode. On comparison of preoperative and intraoperative response amplitudes with regard to stimulus polarity, tympanic electrocochleography appears to be a useful method of gathering preliminary information on the status of the patient's auditory system. In this study, tympanic ECOG was found to have some predictive value when trying to ascertain the best intraoperative monitoring situation. PMID- 8424479 TI - Translabyrinthine approach to acoustic tumors. PMID- 8424480 TI - Perfusion response during electrical stimulation of the auditory nerve in profoundly deaf patients: study with single photon emission computed tomography. AB - Brain activation procedures associated with single photon emission computed tomography (SPECT) recently have been developed in healthy controls and in diseased patients to help in their diagnosis and treatment. We investigated the effects of an electrical stimulation of the auditory nerve on the cerebral distribution of 99mTc-HMPAO in seven profoundly deaf adults. The promontory test was positive (PT+) in six patients and negative (PT-) in one. A count increment of the temporal cortex was observed in all patients, and was higher in all patients with PT+ (right 20%, left 19%) than in the patient with PT- (right 9%, left 3%). These results led us to consider 99mTc-HMPAO brain SPECT imaging as a potential preoperative test for cochlear implant. It might be helpful for the evaluation of cortical perfusion response to an electrical stimulation. Sensitivity tests for the method are requested before this clinical application. PMID- 8424481 TI - Intramuscular streptomycin effect on dark cells of utricle in guinea pigs. AB - Streptomycin has been used in the treatment of Meniere's disease for almost half a century. Clinical studies showed that streptomycin may eliminate vertigo attacks and stabilize or improve hearing in the majority of patients. Animal experiments have demonstrated severe damage to the vestibular hair cells after streptomycin treatment. This study is to observe the effect of intramuscular streptomycin on dark cells of utricle. Guinea pigs that had received intramuscular injections of streptomycin sulfate 400 mg/kg per day for 1 and 2 weeks were studied. The dark cells of utricle were observed under transmission electron microscope. The pinocytotic vesicles and rough endoplasmic reticula markedly decreased and the plasmalemma infoldings in the lower part of the cell reduced in the 1-week group. The luminal membrane of the cells bulged out on the surface. After the cell membrane ruptured, the cytoplasmic organelle moved into the endolymphatic space and the cell dissolved. The morphologic changes indicated that streptomycin damaged the cytoplasmic granules and the plasma membrane infoldings of the dark cells. These cytologic characteristics are engaged in fluid secretion. The damage of secretory function of the dark cells may reduce the volume of endolymph. PMID- 8424482 TI - Apparent apical endolymphatic hydrops: computer-aided three-dimensional reconstruction and histologic study of the apical turn of the cochlear duct. AB - Computer-aided three-dimensional reconstruction of the apex of a cochlea and microscopic evaluation of 14 clinically and histologically normal temporal bones were used to demonstrate that what appears to be distention of Reissner's membrane is normal anatomy. The apparent distention is the transition area between the circular insertion of the apex of the cochlear duct and the rest of Reissner's membrane, which is straight. PMID- 8424483 TI - Surgery for epitympanic cholesteatoma: evaluation of training and experience. AB - Results of surgery in a series of 175 ears with attic cholesteatoma were studied in relation to the experience of the surgeons. Residents receive an annual course in temporal bone dissection and are given daily surgical instruction in the operating theater. Their performance as to hearing results, healed tympanic membrane, postoperative discharge, and number of recurrences were statistically not different from those of the faculty. The method of intact canal skin surgery is described in detail because training in this type of surgery develops skill in soft tissue conservation. Continuous clinical instruction and frequent evaluation of progress are necessary to guarantee specialist-level results. PMID- 8424484 TI - Acoustic neuroma and pregnancy. AB - From January 1978 through July 1989, 360 patients underwent primary removal of an acoustic neuroma at the Mayo Clinic. Seventy of these patients were women under the age of 46 years; six of them were pregnant between the time of onset of their symptoms and the removal of their tumor. A review of the records of these six women revealed the presenting complaint in most to be central neurologic symptoms rather than labyrinthine symptoms. All had unilateral tumors. These tumors tended to be larger and more vascular than those in the nonpregnant cohort, and to have greater surgical morbidity. The pure-tone hearing levels were normal in four of these patients. All women experienced normal vaginal delivery without complication. Based on our experience with these women, we believe that acoustic neuroma present during pregnancy may be managed by close observation until the postpartum period, provided there are no impending neurologic complications. PMID- 8424485 TI - Vestibular ataxia following shuttle flights: effects of microgravity on otolith mediated sensorimotor control of posture. AB - Orbital spaceflight exposes astronauts to an environment in which gravity is reduced to negligible magnitudes of 10(-3) to 10(-6) G. Upon insertion into earth orbit, the abrupt loss of the constant linear acceleration provided by gravity removes the otolith stimulus for vestibular sensation of vertical orientation constantly present on Earth. Since the central nervous system (CNS) assesses spatial orientation by simultaneously interpreting sensory inputs from the vestibular, visual, and proprioceptive systems, loss of the otolith-mediated vertical reference input results in an incorrect estimation of spatial orientation, which, in turn, causes a degradation in movement control. Over time, however, the CNS adapts to the loss of gravitational signals. Upon return to Earth, the vertical reference provided by gravitational stimulation of the otolith organ reappears. As a result, a period of CNS readaptation must occur upon return to terrestrial environment. Among the physiological changes observed during the postflight CNS readaptation period is a disruption of postural equilibrium control. Using a dynamic posturography system (modified NeuroCom EquiTest), 16 astronauts were tested at 60, 30, and 10 days preflight and retested at 1 to 5 hours, and 8 days postflight. All astronauts tested demonstrated decreased postural stability immediately upon return to Earth. The most dramatic increases in postural sway occurred during those sensory conditions in which both the visual and proprioceptive feedback information used for postural control were altered by the dynamic posturography system, requiring reliance primarily upon vestibular function for control of upright stance. Less marked but statistically significant increases in sway were observed under those conditions in which visual and foot support surface inputs alone were altered.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424486 TI - Perforated tympanic membrane after blast injury. AB - The rate of spontaneous healing of tympanic membrane perforations in relation to time after blast injury, size, and location of perforations, was evaluated in 147 patients (210 ears). In 46 patients a paper patch was placed over the perforation. Spontaneous healing was seen in 155 ears (74%), 131 (62%) healed within the first 3 months and 145 ears (69%) healed within 10 months. Large and central kidney-shaped perforations had the least tendency to heal and usually required surgical intervention. Tympanoplasty was performed on 33 ears (15%), ossiculoplasty on 5 and mastoidectomy on 10. Intervention is recommended when the tympanic membrane remains unhealed for at least 10 months. PMID- 8424487 TI - Negative observation of intravenously administered fluorescein in perilymph at stapes surgery. PMID- 8424488 TI - Reanimation options in facial paresis following traumatic birth injury. PMID- 8424489 TI - Recurrent alternating facial palsy in a child. PMID- 8424490 TI - Left tympanic segment facial nerve neuroma and hearing loss, but normal facial function. PMID- 8424491 TI - Facial hyper- and hypokinesia following face lift surgery. PMID- 8424492 TI - Neurologic dysfunction after CAB or valvular surgery: is the medium the miscreant? PMID- 8424493 TI - General anesthetic action: an obsolete notion? PMID- 8424494 TI - Sedation scales: measures of calmness or somnolence? PMID- 8424495 TI - Central nervous system complications after cardiac surgery: a comparison between coronary artery bypass grafting and valve surgery. AB - Central nervous system (CNS) complications (disturbance of consciousness, focal motor deficits, and seizures) after coronary artery bypass grafting (CABG) and cardiac valve surgery were studied retrospectively. The incidence of CNS complications was significantly more frequent in CABG (11%, 71/638) than in valve surgery (7%, 24/345). Major contributory factors of CNS complications were preexisting cerebrovascular disease and cardiopulmonary bypass time. In comparison to previous reports, older age, hypertension, diabetes mellitus, and cerebrovascular disease were more common in the patients undergoing CABG. The preexisting cerebrovascular disease and prolonged cardiopulmonary bypass time probably increase the risk of cerebral embolism and/or cerebral hypoperfusion. We conclude that patients undergoing CABG surgery are at greater risk for neurological damage in comparison to those undergoing valve surgery. PMID- 8424496 TI - Quality assurance for intraoperative transesophageal echocardiography monitoring: a report of 846 procedures. AB - We evaluated our experience with 846 consecutive transesophageal echocardiography (TEE) intraoperative monitoring procedures performed between November 1989 and July 1991. TEE frequency was 36 +/- 11 per month (range 16-55) and represented 69.8% of cardiac valve surgery cases, 40.2% of coronary artery bypass graft cases, and 2.2% of total operative caseload. Major patient complications consisted of transient vocal cord paresis and ingestion of glutaraldehyde disinfectant solution. Minor complications consisted of a chipped tooth (one case) and pharyngeal abrasions (three cases). The Quality Assurance (Q/A) Program evaluated both record keeping and quality of imaging, as judged by cardiologist echocardiographer reviewers. The percentage of completion for each Q/A indicator was as follows: medical record documentation, 88%; database form annotation, 94%; and provision of videotape recording, 91%. TEE database forms were analyzed further in terms of the percentage of fields completed. Completion scores were 73%. The following scoring system was utilized for videotape evaluation by the cardiologists: 1 = excellent; 2 = good; 3 = poor. The median grade for both two dimensional echocardiography and color flow Doppler (CFD) examinations was 2. Poor quality images (grade 3) were present in 15.2% of two-dimensional echocardiography and 20.3% of color flow Doppler examinations, and disproportionately associated with 4/26 attendings. Supplemental audit of the cardiology reviewers performance demonstrated 569/846 videotapes showed no objective evidence of review. The cardiology reviewer forms of the remaining 277 videotapes were evaluated in terms of the percentage of fields completed. The completion score was 56%. These data suggest the need for formal Q/A for intraoperative TEE, both for anesthesiologists and reviewing cardiologists. PMID- 8424497 TI - Influence of different glucose-insulin-potassium regimes on glucose homeostasis and hormonal response in cardiac surgery patients . AB - Intravenous infusion of glucose/insulin in combination with potassium (GIK) is an often used technique to improve myocardial preservation in cardiac surgery. In a randomized study in 50 patients undergoing elective aortocoronary bypass grafting with good ventricular function, the influence on glucose homeostasis and hormonal response to four different glucose/insulin regimes were compared to an untreated control: 1) 50 g of glucose mixed with 100 U of human insulin (HI/100); 2) 50 g of glucose mixed with 100 U of bovine insulin (BI/100); 3) 50 g of glucose mixed with 50 U of human insulin (HI/50); and 4) 50 g of glucose mixed with 50 U of bovine insulin (BI/50) [corrected]. Glucose/insulin were given in combination with 70 mmol of potassium 40 min before beginning the operation. In addition to blood glucose concentrations various endocrine variables were studied before, during and after cardiopulmonary bypass (CPB). Hemodynamic data (arterial blood pressure, heart rate, cardiac index, systemic vascular resistance) were similar in all five groups. Inotropic support after CPB was necessary in none of the patients. Blood glucose levels showed no differences among all GIK groups (groups 1-4) but were significantly higher than in the control patients. The incidence of severely reduced (< 50 mg/dL) or elevated blood glucose level (> 300 mg/dL) did not differ between HI- and BI-treated patients but was significantly lower in the untreated control. Insulin plasma level increased significantly after infusion of GIK with higher levels in HI- than BI-treated patients (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424498 TI - Cardioprotection by superoxide dismutase: a catecholamine-dependent process? AB - Oxygen-derived free radicals may contribute to tissue injury in myocardial ischemia although the mechanism is unclear. Catecholamines possibly could be involved in the genesis of free radicals because it has been demonstrated that oxygen free radicals may be generated by autooxidation of noradrenaline. Superoxide dismutase (SOD) protects the myocardium against injury by superoxide anion radicals. We, therefore, examined whether the cardioprotective effect of superoxide dismutase still could be demonstrated after depletion of catecholamine stores by reserpine (7 mg/kg intraperitoneally 24 h premortem). We used electrically paced isolated hearts perfused according to Langendorff (Tyrode's solution, Ca2+ 1.8 mmol/L, constant perfusion pressure: 70 cm H2O, 3 Hz). Myocardial ischemia was induced by occlusion of a left coronary artery branch. Epicardial NADH-fluorescence was used for quantitation of the myocardial ischemia. SOD (48 U/mL) did not influence global coronary flow or left ventricular pressure significantly (P > 0.05). In control hearts, SOD significantly diminished both size and intensity of epicardial NADH-fluorescence after repetitive coronary ligatures (-45%) (P < 0.05). In hearts with depleted catecholamine stores, this cardioprotection by SOD was no longer observed (P > 0.05). Stimulation of noradrenaline overflow by increasing the pacing rate of control hearts from 180/min up to 300/min after coronary occlusion also significantly enlarged myocardial ischemia (P < 0.05). This pacing rate-dependent growth of myocardial ischemia could be prevented completely by either prior depletion of catecholamine stores with reserpine or SOD. Therefore, noradrenaline seems to be the most important source for the generation of oxygen free radicals during myocardial ischemia in isolated saline-perfused rabbit hearts. PMID- 8424499 TI - Evaluation of sufentanil anesthesia obtained by a computer-controlled infusion for cardiac surgery. AB - To determine the "therapeutic window" for sufentanil in cardiac surgery, nine hemodynamically stable, lightly premedicated patients were anesthetized with sufentanil to undergo coronary artery bypass grafting. Sufentanil was administered by a computer-controlled infusion pump programmed with an infusion scheme based on average sufentanil pharmacokinetics. The presence of somatic or hemodynamic responses to tracheal intubation, skin incision, sternotomy, and during electrocauterization, to isolate the internal mammary artery were correlated with plasma concentrations of sufentanil. The concentration versus response/no response relationships were analyzed by application of the logistic (Hill) equation. The CP50 of sufentanil to suppress responses to intubation, incision, and sternotomy was 7.1 ng/mL and for mediastinal dissection it was 12.7 ng/mL. Interpatient variability was as much as that reported previously for other opioids in other types of surgical patients. The computer-controlled infusion scheme consistently produced sufentanil plasma concentrations in excess of the target concentrations, but there was a strong correlation between target and actual sufentanil concentrations (r = 0.88, P < 0.05). PMID- 8424500 TI - Myocardial perfusion following acute subarachnoid hemorrhage in patients with an abnormal electrocardiogram. AB - To test the hypothesis that acute subarachnoid hemorrhage is associated with abnormal myocardial perfusion, we assessed myocardial blood flow with thallium scintigraphy in 19 patients with a confirmed subarachnoid hemorrhage and an abnormal electrocardiogram. A thallium scan was performed at the bedside of each patient 3 +/- 2 days (mean +/- SD) after subarachnoid hemorrhage and subsequently was analyzed both qualitatively and quantitatively. Patients averaged 58 +/- 13 yr of age and 68% had one or more cardiac risk factors. The neurologic condition of patients on the day of the scan was II (median; range I-V) on the standard 5 point scale of Botterell. Abnormalities on a standard 12-lead electrocardiogram obtained on the same day as the scan consisted of repolarization changes in most patients; 10 had T wave inversions and 8 had nonspecific ST segment changes. Thirty-two percent (n = 6) of patients had an abnormal thallium scan. There were, however, no features of the clinical history, electrocardiogram pattern, or neurologic condition that were associated with a positive scan. For instance, 2 of 4 patients with diffuse deeply inverted T waves had a normal thallium scan, whereas the scan was abnormal in 2 of 8 patients with minor nonspecific electrocardiographic abnormalities. The thallium scan was also positive in neurologically intact (grade I) as well as severely injured (grade V) patients. Thus, abnormal myocardial perfusion and possibly myocardial ischemia occur frequently following subarachnoid hemorrhage, but no specific electrocardiographic characteristic identifies patients with a perfusion abnormality. PMID- 8424501 TI - Physiologic and anesthetic alterations on spinal-sciatic evoked responses in swine. AB - Spinal-sciatic evoked responses (ScER) have been used successfully to monitor the integrity of the anterior spinal cord during spinal surgery. To evaluate the effects of hypercarbia, hypocarbia, induced hypotension, and hypothermia on the ScER, ten swine anesthetized with ketamine were subjected to varying levels of PaCO2, hypothermia, and induced hypotension. During variation of one physiologic variable, the other variables were closely regulated. There were no significant changes associated with variations in PaCO2. Decreasing temperature provided a consistent increase in latency (r = -0.78, P < 0.001) with no significant alteration in amplitude. Graded hypotension caused little increase in latency (3.2% at 30 mm Hg). The amplitude decrease averaged 23% at 60 mm Hg with a maximal decrease of 50% at 30 mm Hg. To study inhaled anesthetics, 21 swine anesthetized with ketamine were subjected to nitrous oxide (50% and 70%). After termination of the nitrous oxide, one of the potent inhaled anesthetics (n = 7 each) was administered in 0.25 minimum alveolar concentration (MAC) increments. Nitrous oxide caused a significant decrease in amplitude (average 43% and 61% at 50% and 70%) with minimal changes in latency. There was a dose-dependent decrease in amplitude and increase in latency with all inhaled anesthetics. The ScER disappeared at 1.0 MAC with all anesthetics. There were no differences between effects of equipotent concentrations of inhaled anesthetics. These findings may be helpful in the interpretation of the ScER response during anesthesia and surgery. PMID- 8424502 TI - Effect of intracarotid and intraventricular morphine on regional cerebral blood flow and metabolism in pentobarbital-anesthetized dogs. AB - Potent opiate analgesics, administered either epidurally, intrathecally, or intravenously, are a common adjunct to pain control in the perioperative period. Little is known, however, of the effects of opiate agonists on cerebral blood flow (CBF) and metabolism. Current studies are complicated by the hypotensive effects of these compounds and their route of administration. To circumvent these difficulties we studied the effects of intraventricular and intracarotid morphine sulfate, in doses that do not affect arterial blood pressure, on regional CBF and total cerebral oxygen consumption (CMRO2) in pentobarbital-anesthetized dogs. Five dogs received 0.04 mg/kg morphine via intracarotid injection. Five additional dogs received 0.2 mg/kg morphine via ventricular cisternal infusion over 5 min, and five control dogs received mock cerebrospinal fluid at the same infusion rate. CBF was measured using the radiolabeled microsphere technique. Intracarotid morphine decreased neurohypophyseal blood flow to 58% of control, but it did not alter blood flow to any other brain region, except caudate nucleus, or cause a change in CMRO2. Infusion of mock cerebrospinal fluid in the cerebral ventricles did not alter CBF or CMRO2. Ventricular-cisternal perfusion of morphine caused a transient increase in CBF (24 +/- 2 to 37 +/- 6 mL min-1 100 kg-1) but had no effect on spinal cord blood flow or CMRO2. Neurohypophyseal blood flow, however, decreased to 40% of control levels (480 +/- 76 to 176 +/- 42 mL min-1 100 kg-1) after 2 min and gradually returned to control levels at 60 min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424503 TI - The effect of halothane and isoflurane on neurologic outcome following incomplete cerebral ischemia in the rat. AB - The relation between sympathetic activity and neurologic outcome was evaluated during fentanyl/nitrous oxide (N2O) (25 micrograms.kg-1.min-1 plus 70% N2O in oxygen), halothane (1.1% inspired), and isoflurane (1.4% anesthesia in a rat model of incomplete cerebral ischemia. Ischemia was produced by right carotid ligation combined with hemorrhagic hypotension to 30 mm Hg for 30 min. Plasma catecholamines were measured during ischemia. Neurologic outcome was measured for 3 days following incomplete ischemia. Both halothane and isoflurane decreased plasma catecholamines 50-80% and improved ischemic outcome compared to fentanyl/N2O anesthesia (P < 0.05). These results indicate a relation between the ability of inhaled anesthetics to decrease sympathetic activity and to improve outcome from incomplete cerebral ischemia. PMID- 8424504 TI - Effects of epidural morphine and intramuscular diclofenac combination in postcesarean analgesia: a dose-range study. AB - To assess the efficacy of combinations of epidural morphine and intramuscular diclofenac in postcesarean analgesia, a double-blind, randomized study was conducted. Epidural anesthesia was administered to 120 parturients who were randomly allocated into six treatment groups; these groups, A, B, C, D, and E, received 0.5, 1, 2, 3, and 4 mg of epidural morphine in 10 mL of normal saline solution, respectively, and 75 mg (3 mL) of diclofenac intramuscularly (IM). Group F received 4 mg of epidural morphine in 10 mL of normal saline solution and 3 mL of normal saline solution IM. Epidural injections were given after delivery of the placenta and IM injections were given on arrival in the recovery room. Verbal analogue pain score and pruritus score were recorded at 2, 4, 8, 12, 18, and 24 h after epidural injection. Subjective scores of overall pain relief also were recorded at 24 h. Results showed that group A had the highest pain scores and required more supplemental meperidine than groups C, D, E, and F. None of the patients in groups D and E requested supplemental analgesia. There were no differences in wound pain score among groups B, C, D, E, and F. Compared with group F, groups D and E experienced a superior analgesic effect in relieving uterine contraction pain from 4 to 12 h (P < 0.05). There were no differences in overall pain relief among the six groups. The incidence of nausea or vomiting or both, pruritus, and bleeding was similar. There was no evident relationship between severity of pruritus and morphine doses. No bradyspnea was observed during the study period.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424505 TI - Time course of sympathetic blockade during epidural anesthesia: laser Doppler flowmetry studies of regional skin perfusion. AB - We studied the time course of sensory and sympathetic blockade in response to epidural local anesthetic test and bolus doses in 11 patients. Sympathetic activity was measured by monitoring cutaneous perfusion in the foot and the reflex vasoconstrictive response to deep inspiration (IGVR) using laser Doppler flowmetry. Sensory tests included the detection of touch, cold and painful stimuli. Following the 3-mL test dose perfusion increased to 192 +/- 38% (mean +/ SEM) of baseline (P < 0.05) in the patients with successful epidural anesthesia (9 of 11). In 8 of these patients, IGVR decreased to 73 +/- 10% of baseline (P < 0.05) within 6 min of the test dose, and preceded changes in sensation to cold, pin-prick, and light touch by 3.8 +/- 3.5 min (P < 0.2), 9.6 +/- 3.1 min (P < 0.01), and 11.6 +/- 2.7 min (P < 0.01), respectively. Five control patients who received only 60 mg of lidocaine intravenously and the two patients with failed epidurals did not show any perfusion or IGVR changes. This study confirms that sympathetic block precedes sensory block in sacral dermatomes after epidural anesthesia. Perfusion and IGVR changes are sensitive measures of sympathetic blockade and may predict successful epidural catheter placement. PMID- 8424506 TI - A comparison of postoperative epidural analgesia between patients with chronic cancer taking high doses of oral opioids versus opioid-naive patients. AB - Our study evaluated 116 surgical patients with cancer who received postoperative epidural analgesia with bupivacaine (BUP) (0.1%) and morphine (MS) (0.01%) during 5 days after epidural-light general anesthesia. Patients in group I (n = 17) were taking opioids in doses larger than 50 mg of morphine daily for 3 mo or more, whereas patients in group II (n = 99) were opioid-naive. Postoperative epidural infusions were started at 10 mL.h-1 for group I and 5 mL.h-1 for group II. All patients were evaluated every 6 h for pain, withdrawal, and overdosing. Dynamic pain scores were kept below 4/10 by titrating infusions and/or giving intravenous (IV) MS 4 mg every hour as needed. Fifteen patients were taking opioids for 3-6 mo and the remainder for more than 6 mo. Mean oral MS preoperative usage for group I was 183 mg (90-360 mg range). All patients experienced adequate analgesia. Group I required more epidural (137 vs 44 mg) and IV (48 vs 10 mg) MS and had a longer requirement for analgesic therapy (9 vs 3 days) when compared with group II. Daily epidural and IV MS usage were always more for group I by two to threefold. No patient experienced respiratory depression or opioid withdrawal during the hospitalization. Thus, epidural BUP-MS appears to provide adequate postoperative analgesia while preventing withdrawal in opioid-dependent patients, if three times the normal dosage and duration of therapy are employed. PMID- 8424507 TI - Time-related differential effects of epidural morphine on the neuraxis. AB - To clarify the site and potency of the analgesic and anesthetic action of epidurally administered morphine, we investigated the effects of epidurally and intravenously administered morphine (100 micrograms/kg) on change in the pressure pain threshold (PPT) and the minimum alveolar concentration (MAC) of halothane. Epidural morphine (EM) increased PPT significantly (P < 0.01) at the points around the surgical incision by 40% and 60% from baseline compared with intravenous morphine (IM) with which PPT remained at baseline 1 and 2 h after administration, respectively. Duration of analgesia was much longer in EM than in IM (18 h vs 1.7 h). EM increased PPT from preoperative value at the forehead and the points around the surgical incision by 9.9% and -24.9% at 1 h, by 10.9% and 3.8% at 4 h, and by -19.5% and -50.4% at 12 h after administration at mean values, respectively. Halothane MAC in EM and IM were 0.54% and 0.57%, respectively, 40 min after administration. Halothane MAC in EM at 4 h and 12 h after administration were 0.45% and 0.70%, respectively. The results suggest that EM provides long-lasting analgesia by its time-related differential effects on the neuraxis. PMID- 8424508 TI - A randomized double-blind comparison of epidural fentanyl infusion versus patient controlled analgesia with morphine for postthoracotomy pain. AB - The authors conducted a prospective, randomized, double-blind comparison of an epidural fentanyl infusion versus patient-controlled analgesia (PCA) with morphine in the management of postthoracotomy pain. Thirty-six patients were randomized into one of two groups. The epidural group received an epidural fentanyl infusion, 10 micrograms/mL, and saline through their PCA machine. The PCA group received an epidural saline infusion and morphine, 1.0 mg/mL, through their PCA device. The infusions were escalated according to a study protocol when pain relief was deemed inadequate by the patients. Pain relief was evaluated by a visual analog pain scale (VAS), both at rest and during coughing, and by verbal rating scores (VRS) of pain relief. Degree of sedation and the frequency of nausea, vomiting, and pruritus were also noted. The VAS, VRS, degree of sedation, and side effects were evaluated every 2 h from 7 AM to 7 PM, for 72 h after surgery. Forced vital capacities were determined before surgery and at 24, 48, and 72 h after surgery. The VAS were significantly lower (P = 0.001), and the Total Pain Relief scores higher (P < 0.02) in the epidural group, signifying better analgesia. There were no differences in postoperative forced vital capacity between the two groups. More patients in the PCA group had greater degrees of sedation on postoperative day 1 (P = 0.005), whereas pruritus was more frequent (P < 0.02) in the epidural group. We conclude that an epidural fentanyl infusion is superior to that of PCA with morphine in the management of pain after thoracotomy. PMID- 8424509 TI - Redistribution of sufentanil to cerebrospinal fluid and systemic circulation after epidural administration in dogs. AB - Due to its higher lipid solubility, sufentanil may be less likely than morphine to migrate rostrally in the cerebral spinal fluid (CSF) and cause delayed respiratory depression following epidural administration. However, early respiratory depression has been reported in patients after relatively large doses of epidural sufentanil. This has been attributed to systemic drug uptake. We used a dog model to investigate the pharmacokinetics and rostral spread of epidural sufentanil in CSF. Sampling catheters were placed in the lumbar subarachnoid space, the cisterna magna, and femoral arteries of six mongrel dogs. Samples of cisternal CSF, lumbar CSF, and blood were drawn at 0, 1, 5, 15, 30, 60, 90, 120, and 180 min after lumbar epidural sufentanil injection. We measured sufentanil concentrations by gas chromatography-mass spectrometry and used the least squares method to a fit tri-exponential function to each sufentanil concentration versus time data set. Paired t-test was used to test for statistical significance. After epidural sufentanil, lumbar CSF concentrations were significantly higher than plasma or cisternal CSF sufentanil concentrations at all assessment times. Sufentanil concentrations were significantly higher in cisternal CSF than in plasma at 30 and 60 min after injection. Sufentanil appeared rapidly in lumbar CSF, reaching a maximum concentration (Cmax) of 57 ng/mL at 6.5 min. In cisternal CSF, a Cmax of 1.2 ng/mL was reached at 21 min, and Cmax in plasma was 0.35 ng/mL at 6 min. The area under the concentration-time curve (AUC) of sufentanil in cisternal CSF was approximately six times higher than the plasma AUC (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424510 TI - Onset, intensity of blockade and somatosensory evoked potential changes of the lumbosacral dermatomes after epidural anesthesia with alkalinized lidocaine. AB - The onset and intensity of blockade of the lumbosacral dermatomes after epidural anesthesia with alkalinized lidocaine were investigated in a randomized, double blind study in 26 patients. Control subjects (n = 13) received 20 mL of 1.37% lidocaine (1.5% lidocaine plus 1 mL saline per 10 mL lidocaine) with added 1:200,000 epinephrine; the solution pH was 6.20 +/- 0.08. Patients in the alkalinized lidocaine group (n = 13) were given 20 mL of 1.37% lidocaine plus added bicarbonate (1 mL sodium bicarbonate per 10 mL 1.5% lidocaine) and 1:200,000 epinephrine; the solution pH was 7.18 +/- 0.10. Posterior tibial nerve (PTN) somatosensory evoked potentials (SSEPs) and L5 and S1 dermatomal SSEPs of both lower extremities were done before and after the epidural. Alkalinization of lidocaine resulted in a significantly shorter time to block the L2, L4, L5, and S1 dermatomes. Motor blockade was significantly more profound in the alkalinized lidocaine group. Thirteen of 78 PTN and L5 and S1 dermatomal SSEPs were abolished in the alkalinized lidocaine group compared to 4 of 78 SSEPs in the nonalkalinized group. Alkalinization of lidocaine is recommended to shorten the time to block the L5-S1 dermatomes when epidural anesthesia is planned for lower extremity surgery. PMID- 8424511 TI - Intraarticular analgesia following knee arthroscopy. AB - A randomized, double-blind, controlled study was conducted in patients undergoing elective knee arthroscopy to assess the analgesic effect of intraarticular morphine and bupivacaine, alone and in combination. Patients in group 1 (n = 10) received 5 mg of morphine in 25 mL of saline; patients in group 2 (n = 10) received 25 mL of 0.25% bupivacaine (62.5 mg); patients in group 3 (n = 10) received a combination of 5 mg of morphine and 62.5 mg of bupivacaine in 25 mL dilution; and patients in group 4 (n = 10) received 25 mL of saline. All the drugs were injected intraarticularly. Postoperative pain was assessed using the visual analogue scale at 1, 2, 4, 8, and 24 h after the intraarticular injection. The need for supplemental analgesia was recorded. Results showed that there was no significant difference in the pain scores or analgesic requirements between groups 1 and 3. Patients in groups 1 and 3 had significantly lower pain scores than those in groups 2 and 4. These low pain scores were associated with lower requirements of supplementary analgesics. The patients in group 4 showed the highest pain scores and analgesic requirements. We conclude that intraarticular morphine significantly reduces postoperative pain following knee arthroscopy and that there is no advantage of combining bupivacaine with morphine. PMID- 8424512 TI - A measure of intraoperative attention to monitor displays. AB - Vigilance is an important but difficult to measure attribute in anesthesia practitioners. We present a modified standard method to assess intraoperative vigilance toward electronic data displays. The response time to detect a simulated abnormal value on the physiologic monitor was measured. Eight anesthesia residents were studied during 60 surgical procedures. Responses to 439 abnormal values were analyzed. The average response time was 61 +/- 61 s (mean +/ SD), and 56% of the detections were made within 60 s. However, 16% of the abnormal values were undetected during the 5 min that they were displayed. Response times and the rate of missed events were greater during induction of anesthesia (a time of high workload) than during the maintenance or emergence phases of anesthesia. Response times were shorter during procedures on ASA 1 patients than on ASA 3 patients. The results suggest that anesthesiologists usually quickly detect abnormal values on physiologic monitors and that less attention is devoted to monitors during periods of high workload. PMID- 8424513 TI - Correlative study of behavior and synaptic events during halothane anesthesia in the lamprey. AB - Behavioral anesthetic effects of halothane were compared with neurophysiological data obtained from monosynaptically coupled, single axon-giant neuron pairs in the isolated central nervous system of the lamprey in vitro. Based on the assumption that the number of halothane molecules at the site of anesthetic action is determined by molar concentration of the bath, depths of anesthesia were characterized in terms of synaptic events. The concentration producing anesthesia in 50% (AC50) and 95% (AC95) of the animals were 0.32 and 0.51 mM, respectively. With these concentrations, the depression of glutamate (GLUT) mediated synaptic excitation remained less than 50%. Maximum suppression of the excitatory synaptic transmission was observed in the presence of 1.7 mM halothane which would be equivalent to the inhalation of 5.5% of this agent at 37 degrees C. At these concentrations, postsynaptic membranes did not respond to the bath applied GLUT agonists. The decrease of agonist-induced responses to almost the same extent as excitatory postsynaptic potential (EPSP) amplitudes in the presence of halothane argues against a presynaptic mechanism affecting transmitter release. Persistent voltage-gated Na+ channel function at concentrations less than 2.4 mM is evidence against the involvement of this mechanism at these levels of anesthesia. The present results suggest that with depths between AC50 and AC95, anesthesia is associated with a partial reduction of sensitivity of the postsynaptic membrane to the excitatory amino acid transmitters. PMID- 8424514 TI - Influence of age on awakening concentrations of sevoflurane and isoflurane. AB - We determined whether age, duration of anesthesia, gender, or type of surgery significantly influenced end-tidal concentrations on awakening from anesthesia with sevoflurane and isoflurane in 39 healthy ASA physical status I patients. Postoperatively, the end-tidal anesthetic concentration was maintained at a constant level at least for 15 min. If patients failed to open their eyes on request, the end-tidal concentration was decreased and again maintained at a constant level for 15 min. The anesthetic concentration midway between the value permitting the response and that just preventing the response was recorded. The end-tidal concentrations on awakening from anesthesia were 0.62 +/- 0.02% (mean +/- SE) for sevoflurane and 0.41 +/- 0.02% for isoflurane. Awakening concentration of sevoflurane and isoflurane correlated significantly with age (P < 0.001), but not with duration of anesthesia, gender, or type of surgery. The authors conclude that awakening concentration decreases at the similar rate of decrease in minimum alveolar concentration (MAC) with increasing age; and therefore, the ratios to MAC are fairly constant, being 0.34 for both sevoflurane and isoflurane. PMID- 8424515 TI - Desflurane versus isoflurane in patients with chronic hepatic and renal disease. AB - Forty patients with chronic hepatic (n = 20) and renal (n = 20) disease were studied randomly for the purpose of evaluating safety and laboratory values. These included alanine and aspartate aminotransferases, alkaline phosphatase, total bilirubin, creatinine, and blood urea nitrogen. Patients with hepatic disease received either desflurane (n = 9) or isoflurane (n = 11) and patients with renal disease received the two anesthetics in equal subsets. In the patients with hepatic and renal disease, there were no differences in laboratory values (P < 0.05). We conclude that, in patients with either hepatic or renal disease, laboratory values from pre- to postoperative state did not change significantly within 24 h of anesthesia, and no differences in these changes existed between desflurane or isoflurane. PMID- 8424516 TI - The effects of glucose on plasma amino acids and pyruvate during upper abdominal surgery. AB - To assess the potential benefits of perioperative glucose administration, the plasma concentrations of insulin, free fatty acids, ketone bodies, amino acids, lactate, and pyruvate were measured in 20 patients undergoing partial gastrectomy. Ten patients received intravenous glucose at 10 g/h during the operation, and the other 10 patients, as control, received no glucose. Plasma glucose, insulin activity, pyruvate and alanine concentrations in the glucose receiving group increased significantly during the operation in comparison with the control group. In contrast, in the glucose-receiving group the plasma ketone bodies and branched-chain amino acids, especially leucine and isoleucine, decreased significantly during surgery. These results suggest that administration of glucose stimulated insulin secretion and resulted in accumulation of the substrates such as pyruvate and alanine which were utilized readily in the body. Glucose administration is useful in suppressing catabolism during upper abdominal surgery. PMID- 8424517 TI - The effect of dopamine on graft function in patients undergoing renal transplantation. AB - We studied the effect of a low-dose dopamine infusion on graft function in 60 patients undergoing transplantation with cadaveric kidneys in a prospective controlled trial. Recipients were allocated to either a control or a dopamine group, the latter receiving a 3 micrograms.kg-1 x min-1 infusion of dopamine starting intraoperatively. Evaluation of dopamine's effect was undertaken in two stages, namely, (i) initial graft function 1 wk after transplantation and (ii) graft survival at 3 mo. Initial graft function was determined by the ability of the transplanted kidney to reduce serum creatinine, and the development of acute tubular necrosis as confirmed by renal biopsy. Of the dopamine group 33.3% developed acute tubular necrosis compared to 23.3% of the control group. The second-stage evaluation was based on plasma creatinine levels and the requirement for dialysis within 3 mo of transplantation. 92.8% of the dopamine group and 76.9% of the control group had good graft function. No statistically significant difference between the two groups was found. The perioperative infusion of dopamine at 3 micrograms.kg-1 x min-1 was not shown to have any beneficial effect on the transplanted kidney in patients who do not have serious vascular disease, or who do not receive kidneys subjected to prolonged hypotension or prolonged preservation or anastomotic times. PMID- 8424518 TI - Effect of temperature on volatile anesthetic depression of myocardial contractions. AB - Anesthetics are studied and used under conditions of hypothermia both experimentally and clinically. To determine if myocardial contractile depression by anesthetics is equivalent at different temperatures, depressant effects on guinea pig papillary muscle contractions were compared at 37 degrees C vs 30 degrees C. Stimulation after rest and at rates from 0.1 to 3 Hz was used. After control measurements were established at one temperature, effects due to equi anesthetic concentrations (equivalent to approximately 0.9 minimum alveolar concentration) of halothane, isoflurane, or enflurane were determined, followed by recovery measurements. The same procedure was then applied at the other temperature. Hypothermia itself had positive inotropic effects: contractions at 30 degrees C developed about twice the tension as at 37 degrees C; the maximum rate of tension development (dT/dt-max) at 30 degrees C was about 50% greater than at 37 degrees C (1-3 Hz stimulation). When anesthetic contractile depression was assessed by the fractional decrease in dT/dt-max, the anesthetics were more depressant at 30 degrees C than at 37 degrees C, an effect which was most prominent at 0.5 Hz. In contrast to the anesthetics, 1 microM ryanodine which decreases sarcoplasmic reticulum (SR) Ca2+ release, and 0.1 microM nifedipine which decreases Ca2+ entry, both depressed dT/dt-max less at 30 degrees C than at 37 degrees C. Furthermore, ryanodine, nifedipine, and a decrease in external [Ca2+] all exhibited distinct rate-dependent contractile depression at 37 degrees C (typically greatest at 2-3 Hz) not seen with anesthetics.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424519 TI - Oral transmucosal fentanyl citrate (OTFC) for the treatment of postoperative pain. AB - Oral transmucosal fentanyl citrate (OTFC) has been used in a variety of clinical situations. This study was designed to determine if OTFC could provide analgesia to patients with acute pain after major surgery. Following written informed consent, 38 ASA Physical Status I-III patients undergoing either a total hip replacement or total knee arthroplasty were studied prospectively. The patients were randomly allocated to receive either OTFC (7-10 micrograms/kg) or a placebo identical in appearance to an OTFC unit. General anesthesia was administered for surgery, and patient-controlled analgesia (PCA) with morphine was initiated in all patients. The PCA interval dose was adjusted to provide adequate analgesia as determined by the patient and physician; the PCA lock-out time was not changed. On the morning after surgery, the most recent 12 h of PCA data (milligrams per hour of morphine and PCA attempts per hour) were recorded. OTFC or placebo units were administered at times 0, 4, and 8 h during a 12-h study, resulting in three identical units being completely consumed. PCA data, as well as incidence and severity of any adverse side effects, were recorded during the study and for the next 12 h. Treatment groups were compared for similarity, and study variables were analyzed. Twenty-eight patients completed the study, 13 in the control group and 15 in the OTFC group. There were no significant differences between the study groups as to patients' age, gender, ASA classification, or surgical procedure. In addition, there were no differences between the groups in the number of PCA attempts or delivered dose of morphine during the prestudy or poststudy periods.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424520 TI - Fentanyl attenuates porcine coronary arterial contraction through M3-muscarinic antagonism. AB - The "antimuscarinic effect" of fentanyl and its dependence on subtypes of receptors were characterized in isolated porcine coronary arteries. Left anterior descending coronary arteries were dissected from the hearts of 60 adult pigs obtained at a slaughterhouse and prepared for isometric tension studies. The effects of fentanyl on the cumulative concentration-response curve for acetylcholine were obtained in the presence and absence of muscarinic blockade by atropine. Fentanyl shifted the concentration-response curve to the right in a concentration-dependent fashion. Atropine shifted the concentration-response curve to the right, and no further shift was caused by fentanyl. To investigate the dependence on muscarinic receptor subtypes, the effect of fentanyl on acetylcholine-induced contraction was examined in the presence of specific M1-, M2-, and M3-muscarinic antagonists. The pA2 values for fentanyl decreased significantly in the presence of atropine (a nonspecific antagonist) and also in the presence of p-F-HHSiD (an M3-antagonist). In contrast, no significant change of pA2 value for fentanyl was observed in the presence of both pirenzepine (an M1 antagonist) and methoctramine (an M2-antagonist). We conclude that fentanyl has an antimuscarinic effect, and that this antagonism occurs in a competitive manner. Furthermore, the significant decrease of the pA2 value for fentanyl in the presence of M3-, but not in the presence of M1 + M2-antagonists, suggests that the attenuation of cholinergic contraction of porcine coronary arteries by fentanyl is mediated through the M3-muscarinic receptor subtype. PMID- 8424521 TI - A comparison of CO2 laser ignition of the Xomed, plastic, and rubber endotracheal tubes. AB - The combustibility of Xomed LaserShield endotracheal tubes was compared to that of polyvinylchloride (PVC) and rubber endotracheal tubes at 15, 17, and 20 watts (W) of power from a CO2 laser. Oxygen, 5.0 L/min, flowed through the tubes during the investigation. The laser was aimed perpendicularly at the shaft of the endotracheal tube under study and actuated until an intraluminal fire occurred. The times to blow torch combustion of the PVC tubes were 1.7, 1.8, and 1.5 s at 15, 17, and 20 W, respectively. For the Xomed tube, the times to combustion were 85.9, 42.5, and 20.3 s at 15, 17, and 20 W, respectively. Intraluminal (blow torch) fires of the rubber endotracheal tubes occurred after 24.5, 25.4, and 21.9 s at 15, 17, and 20 W. The times to combustion of the PVC and Xomed tubes were significantly different at 15 and 17 W. The times to intraluminal combustion of the rubber tubes were significantly different from those of the Xomed tubes only at 15 W. We conclude that the Xomed tube does not offer consistently better resistance to laser-induced combustion than the PVC or rubber endotracheal tubes tested. PMID- 8424522 TI - "Do not resuscitate" (DNR) orders and the anesthesiologist: a survey. AB - Anesthesiologists were surveyed to determine their experience and opinions regarding "Do Not Resuscitate" (DNR) orders in the perioperative period. Four hundred fifteen questionnaires were mailed and 193 (47%) were returned. One hundred sixty-one (87%) of 186 respondents had been requested to provide (and more than two-thirds had provided) monitored anesthesia care, regional anesthesia, or general anesthesia to a patient with a DNR order. Almost two thirds of the respondents assume DNR suspension in the perioperative period and only half discuss this assumption with the patient/guardian. Less than 50% of respondents would require DNR suspension for a palliative procedure contrasted with > 60% for an elective procedure. After agreeing to a patient's decision to retain their DNR status, > 67%, > 58%, < 49%, and < 33% would utilize positive pressure ventilation with a mask, vasoactive drugs, endotracheal intubation, or defibrillation, respectively, in the event of a cardiopulmonary arrest in the perioperative period. These findings suggest much ambiguity regarding DNR orders in the perioperative period. Further discussion among physicians and patients is warranted. PMID- 8424523 TI - Management of pain in the cancer patient. AB - The pain experience of the cancer patient is the result of many factors, including nociceptive sources, specific pain syndromes, and behavioral contributions. Careful evaluation of the patient is necessary to identify the contributors to the patient's pain experience and to select treatment modalities which address the underlying causes. For patients who are experiencing poorly controlled pain as a result of cancer, therapy often includes multiple management strategies involving more than one discipline. Therefore, an interdisciplinary approach may be more useful for pain management. Disciplines and specialties. involved in such care commonly include anesthesiologists, oncologists, psychiatrists, psychologists, physical therapists, pharmacists, nurses, and social workers. The locus of control often influences how patients respond to their physicians' advice. Patients with a strong internal locus of control usually want to participate actively in treatment decisions. Such patients often resent having decisions made about their treatment without their participation. A lack of sense of control can exacerbate such patients' pain and limit compliance with recommended treatments. Drug therapy is the mainstay of cancer pain management. The therapy should be individualized to the patient, and medications should be selected for specific indications. The WHO three-step analgesic ladder should be used as a guide in selecting analgesics. Drugs should be administered by mouth unless it is impossible to do so, and drug costs should be considered when selecting analgesic medications. Doses should be titrated to response. Adjuvant drug therapy should be considered early and implemented when indicated. Practitioners should be familiar with the medications prescribed and be alert for the appearance of adverse side effects. Patients should be monitored and reassessed continuously. A thorough diagnostic work-up should be completed for new symptoms when indicated. For patients with specific pain syndromes, or for whom drug therapy has not been successful, local anesthetic and neurolytic block therapy and more invasive drug delivery systems (e.g., epidural catheters) should be considered. Although cure may not be attainable in many cancer patients, the obligations of health professionals to these patients are no less than to patients for whom a cure is achievable. Effective pain management has a profound impact on the quality of life, and may give the patient the opportunity to face death with dignity and reduced suffering. PMID- 8424524 TI - Ketorolac-induced bronchospasm. PMID- 8424525 TI - Ketorolac, nasal polyposis, and bronchial asthma: a cause for concern. PMID- 8424526 TI - Severe anaphylactic shock after atracurium. PMID- 8424527 TI - Dyskalemic periodic paralysis and myotonia. PMID- 8424528 TI - Staged segmental scoliosis surgery during regional anesthesia in high risk patients: a report of six cases. PMID- 8424529 TI - Bilateral tension pneumothorax caused by a blocked bacterial filter in an anesthesia breathing circuit. PMID- 8424530 TI - Pneumothorax complicating the use of a Univent endotracheal tube. PMID- 8424531 TI - An alternative to tracheostomy following transsphenoidal hypophysectomy in a patient with acromegaly and sleep apnea. PMID- 8424532 TI - Face mask fitting for edentulous patients. PMID- 8424533 TI - Helium and gas flow. PMID- 8424534 TI - Drager ventilator failure on changing the respiratory rate setting. PMID- 8424535 TI - Does phenylephrine constrict coronary arteries during isoflurane-induced hypotension in dogs? PMID- 8424536 TI - Saline versus water for epidural injection. PMID- 8424537 TI - The Easicap CO2 detector. PMID- 8424538 TI - A proposed fiber-optic scoring system to standardize the assessment of laryngeal mask airway position. PMID- 8424539 TI - A special mask for teaching fiber-optic intubation in pediatric patients. PMID- 8424540 TI - Renal function during infrarenal aortic cross-clamping. PMID- 8424541 TI - Continuous jugular venous oximetry. PMID- 8424542 TI - Negative inotropic effects of propofol as evaluated by the regional preload recruitable stroke work relationship in chronically instrumented dogs. AB - BACKGROUND: Propofol anesthesia often is associated with marked decreases in arterial blood pressure. Previous investigations in vivo have provided conflicting reasons for this clinical finding, including propofol-induced decreases in preload or afterload and/or direct myocardial depressant effects. Interpretation of the results of these studies is complicated by use of indices of myocardial contractility that may only indirectly indicate changes in inotropic state or are significantly dependent on ventricular loading conditions. METHODS: Eight experiments were performed using dogs chronically instrumented for measurement of aortic and left ventricular pressure, the peak rate of increase of left ventricular pressure (dP/dtmax), subendocardial segment length, intrathoracic pressure, and cardiac output. Myocardial contractility was evaluated in conscious and anesthetized dogs using the preload recruitable stroke work (PRSW) relationship, a sensitive, easily quantified, and relatively load independent index of contractile function in normal canine myocardium in vivo. The relationship was derived from ventricular pressure-segment length loops generated by abrupt vena caval constriction. Respiratory variation in ventricular pressure was reduced by calculation of transmural pressure via instantaneous subtraction of intrathoracic pressure from corresponding left ventricular pressure. Systemic hemodynamics and myocardial contractility were recorded and evaluated in the conscious state and after a bolus of 5 mg/kg and a propofol infusion for 15 min at 15, 30, 60, and 120 mg.kg-1.h-1. RESULTS: A significant (P < .05) and dose-dependent decrease in PRSW slope (106 +/- 7 during control to 54 +/- 3 mmHg at the 120 mg.kg-1.h-1 infusion) was observed, demonstrating a direct depression of contractility. Concomitant decreases in left ventricular dP/dtmax and percent segment shortening also were observed. In addition, a significant decrease in systemic vascular resistance occurred at the two largest infusions. CONCLUSIONS: The results indicate that the significant decrease in systemic arterial blood pressure observed during continuous propofol anesthesia in dogs is a result of direct negative inotropic actions of propofol along with its direct effects upon arterial and venous vascular tone. PMID- 8424543 TI - Is ketamine or its preservative responsible for neurotoxicity in the rabbit? AB - BACKGROUND: Although ketamine has been administered spinally in humans, previous neurotoxicity studies have shown that it can induce spinal cord lesions in various animal models. The aim of this work was to evaluate by histologic and blood-brain barrier studies whether different components of the commercial ketamine solution might be responsible for the microscopic lesions observed. METHODS: Forty white New Zealand rabbits were randomly assigned to four groups of 10. One-percent preservative-free ketamine (0.3 ml), 1% d-ketamine, 0.05% chlorobutanol, and 1% lidocaine were intrathecally injected through the atlantooccipital membrane. Laminectomy was performed on day 8, and the dura was preserved using paraformaldehyde-glutaraldehyde fixative. Light and fluorescence microscopy were performed on transverse spinal cord sections by a neuropathologist unaware of injected agents used. Specimens were then graded as normal or abnormal as compared with a control group receiving lidocaine. RESULTS: Isomers of ketamine did not induce spinal cord lesions in either study, but chlorobutanol (the preservative used in the ketamine solution) induced significant severe spinal cord lesions in both studies. CONCLUSIONS: The appearance of spinal cord lesions after intrathecal chlorobutanol strongly suggests that this preservative is responsible for apparent toxicity of ketamine and therefore should not be used in any solution intrathecally injected into humans. PMID- 8424544 TI - A technique for approximately maintaining constant plasma levels of intravenous drugs. AB - BACKGROUND: There is increasing interest among anesthesiologists in the use of continuous infusion of intravenous drugs. The therapeutic effect of most drugs is a function of the concentration at the site of drug effect, which in turn is determined by the plasma concentration. Constant plasma concentrations can be maintained by computer-controlled infusion pumps. However, such equipment is not yet widely available and will be expensive. METHODS: A technique is presented to enable the anesthesiologist to maintain approximately a desired plasma concentration after an arbitrary bolus dose by using a series of infusions with rates decreasing in a stepwise fashion. The algorithm is based on approximating the exact infusion needed to maintain the target plasma concentration by producing this concentration at discrete, specific times. Equations are derived for calculating the sequential rates of the infusion scheme. The equations assume linear pharmacokinetics, and the starting point for derivation of the equations is the assumption that the plasma concentration is given by the convolution of the drug infusion and the unit dose-response function. RESULTS: The accuracy of the technique was assessed by simulating the infusion of fentanyl and midazolam. By using an infusion scheme of three steps, the error was no greater than 38% for fentanyl and no greater than 10% for midazolam. CONCLUSIONS: Other than the assumption of linear kinetics, the algorithm is independent of pharmacokinetic models. Implementation does not require computer-based numerical analysis. PMID- 8424545 TI - The effects of morphine, MK-801, an NMDA antagonist, and CP-96,345, an NK1 antagonist, on the hyperesthesia evoked by carageenan injection in the rat paw. AB - BACKGROUND: The spinal mechanisms underlying the hyperesthetic state during inflammation are little understood. To gain a better understanding of these mechanisms, this study evaluated the effects of intrathecal morphine; MK-801, an N-methyl-D aspartic (NMDA) antagonist; and CP-96,345, an NK1 antagonist, on the hyperesthesia observed after carageenan injection of the rat paw. METHODS: In rats injected with 2 mg carageenan, the paw withdrawal latency (PWL) for the injected paw was typically 5-6 s less than that for the untreated paw, at 2 h after the carageenan injection. Drugs were administered 2 h after the carageenan injection. The magnitude of hyperesthesia was evaluated with the difference score (DS), which was calculated by subtracting the PWL of the untreated paw from the PWL of the injected paw. RESULTS: Intrathecal morphine increased PWLs of both the injected and the untreated paws equally in a dose-dependent manner, but intrathecal morphine did not affect the level of DS. Intrathecal MK-801 increased PWLs of the injected paw to the level of the untreated paw in a dose-dependent manner and increased the DS levels. Intrathecal CP-96,345 had no effect on PWLs of either the injected or the untreated paw. Coadministration of MK-801 with morphine reduced the DS for each dose of morphine. CONCLUSIONS: These data indicate that (1) an NMDA receptor, but not an NK1 receptor, plays an important role in maintaining the hyperesthesia after carageenan injection; and (2) NMDA antagonism has a simple additive interaction with morphine in the carageenan model of inflammatory hyperesthesia. PMID- 8424546 TI - The effect of acute hypocapnia on local cerebral blood flow during middle cerebral artery occlusion in isoflurane anesthetized rats. AB - BACKGROUND: Because the effect of hypocapnia on distribution of cerebral blood flow during focal cerebral ischemia is controversial, this investigation was performed in rats to determine whether hypocapnia, instituted immediately after the onset of focal cerebral ischemia, produces a favorable redistribution of blood flow (an "inverse steal") toward the ischemic territory. METHODS: After surgical preparation during normocapnic isoflurane anesthesia, middle cerebral artery occlusion (MCAO) was performed. Animals were randomized to either immediate institution of hypocapnia (n = 9; PaCO2 23 +/- 2 mmHg) or continued normocapnia (n = 8; PaCO2 40 +/- 2 mmHg). Thirty minutes thereafter, local cerebral blood flow (1-CBF) was determined autoradiographically using 14C iodoantipyrine. Local cerebral blood flow was determined in four coronal brain sections spanning the distribution of the middle cerebral artery. For the hemisphere ipsilateral to MCAO, the areas of cortex in which CBF fell within three specified CBF ranges (0-6, 6-15, and 15-23 ml/100 g/min) were measured and expressed as a percentage of the total area of cortex in that section. In the hemisphere contralateral to MCAO, to confirm the presence of normal CO2 reactivity in non-ischemic brain in this model, average 1-CBF was determined for the cortex, the subcortex, and the entire hemisphere in each coronal section. RESULTS: Hypocapnia resulted in significantly lower 1-CBF in the cortex, subcortex, and entire hemisphere in all coronal sections of brain contralateral to MCAO. In the hemisphere ipsilateral to MCAO, a favorable redistribution of CBF was not observed. For the three more anterior coronal sections (1-3), a significantly larger percentage of the cortex had 1-CBF in the range of 15-23 ml x 100 g-1 x min-1 in the hypocapnia group animals. In sections 2 and 3, significantly larger areas of cortex had 1-CBF in the range of 6-15 ml x 100 g-1 x min-1 in the hypocapnia group than in the normocapnia group. For all sections, there were no significant differences between hypocapnic and normocapnic groups in the area of cortex with 1-CBF in the range of 0-6 ml x 100 g-1 x min-1. CONCLUSIONS: The current study does not provide evidence for the occurrence of a hypocapnia-induced inverse steal phenomenon during acute focal cerebral ischemia of 30 min duration in the rat. The data suggest that, rather than reducing the area of the critically ischemic brain, hypocapnia may increase the size of the region at risk. The data do not support the use of hypocapnia as a therapeutic measure to produce a favorable redistribution of CBF during focal cerebral ischemia of acute onset. PMID- 8424547 TI - Reversal of volatile anesthetic-induced depression of myocardial contractility by extracellular calcium also enhances left ventricular diastolic function. AB - BACKGROUND: Volatile anesthetics depress global left ventricular function by altering intracellular calcium (Ca2+) homeostasis at several sites within the myocyte. Although extracellular Ca2+ partially reverses the negative inotropic effects of volatile anesthetics, the actions of extracellular Ca2+ on anesthetic induced diastolic dysfunction are unexplored. This investigation examined and compared the direct effects of extracellular Ca2+ on left ventricular systolic and diastolic function in conscious and anesthetized dogs. METHODS: Experiments were conducted in the presence of pharmacologic blockade of the autonomic nervous system because autonomic nervous activity may significantly influence the hemodynamic actions of anesthetics and Ca2+ in vivo. Three groups comprised a total of 27 experiments conducted using nine dogs chronically instrumented for measurement of aortic and left ventricular pressure, left ventricular dP/dt, subendocardial segment length, and cardiac output. Myocardial contractility was evaluated using the preload recruitable stroke work relationship slope (Mw). Diastolic function was assessed using a time constant of isovolumic relaxation (tau), a regional chamber stiffness constant (Kp), and maximum segment lengthening velocity during rapid ventricular filling (dL/dtE) and atrial systole (dL/dtA). On 3 separate days, a CaCl2 infusion at 1.25, 2.5, or 5 mg.kg-1 x min-1 was administered. Hemodynamics and ventricular pressure-length loops were recorded after a 20-min equilibration at each dose in the conscious state or during halothane or isoflurane anesthesia. RESULTS: In conscious dogs, CaCl2 produced a significant (P < .05) and dose-dependent increase in contractility as evaluated by Mw. In the presence of halothane anesthesia, CaCl2 increased contractility (Mw of 26 +/- 5 mmHg to 78 +/- 10 mmHg during the high dose of CaCl2), enhanced isovolumic relaxation (tau of 57.9 +/- 4.2 ms to 41.1 +/- 1.9 ms during the high dose of CaCl2), improved rapid ventricular filling (dL/dtE of 11.8 +/- 1.4 mm/s to 20.2 +/- 1.6 mm/s during the high dose of CaCl2), and reduced regional chamber stiffness (Kp of 0.70 +/- 0.18 mm-1 to 0.38 +/- 0.04 mm 1 during the high dose of CaCl2). Similar findings were observed when CaCl2 was administered to dogs anesthetized with isoflurane. CONCLUSIONS: Although CaCl2 produced positive inotropic effects in both the conscious and anesthetized states, CaCl2 did not alter diastolic function in conscious dogs. In contrast, CaCl2 reversed halothane- and isoflurane-induced negative lusitropic actions. The results of the present investigation suggest that improvement of left ventricular performance by CaCl2 during volatile anesthesia may be related to actions in diastole as well as systole. PMID- 8424548 TI - Treatment of postoperative nausea and vomiting after outpatient surgery with the 5-HT3 antagonist ondansetron. AB - BACKGROUND: Postoperative nausea and vomiting following outpatient surgery can significantly delay discharge. This study evaluates the safety and efficacy of ondansetron (a new 5-HT3 antagonist) in the treatment of postoperative nausea and vomiting in patients following outpatient surgery. METHODS: Five hundred outpatient surgical patients (53 male and 447 female), receiving general endotracheal anesthesia, were studied at ten centers. Patients were stratified by gender and received, in a randomized, double-blind manner, 1, 4, or 8 mg ondansetron or placebo in response to nausea and/or vomiting postoperatively. Episodes of vomiting, nausea scores, adverse events, vital signs, and laboratory values were evaluated before and during the 24 h after study drug administration. RESULTS: Complete response to study medication (no vomiting and/or retching, and no rescue antiemetic over the initial 0-2-h period) was more frequent in the ondansetron groups (1 mg 57%, 4 mg 61%, and 8 mg 57%) than in the placebo group (30%, P < .001). For the 0-24-h study a complete response occurred in only 15% of the placebo group compared to 41%, 47%, and 47% of the 1-, 4-, and 8-mg ondansetron groups, respectively (P < .001 for all comparisons with placebo). Median nausea scores (range 0-10) during the initial observation period (0-2 h) were significantly lower for all doses of ondansetron (1.3, 0.8, 1.8 for 1, 4, and 8 mg, respectively) as compared with placebo (2.3). No significant differences occurred in hemodynamic stability, incidence of adverse events, or changes in laboratory values in the ondansetron groups compared to the placebo group. CONCLUSIONS: Ondansetron, in doses less than 8 mg, is a safe, effective antiemetic for treating postoperative nausea and vomiting. PMID- 8424549 TI - Effect of midazolam infusion and flumazenil administration on epinephrine arrhythmogenicity in dogs anesthetized with halothane. AB - BACKGROUND: Midazolam is being selected increasingly for use in patients with cardiovascular compromise. Although clinical doses of midazolam have minimal effects on cardiac function, the influence of midazolam (and other benzodiazepine sedatives) on cardiac arrhythmogenesis has yet to be elucidated fully. METHODS: In this study, we investigated the effect of midazolam, with and without flumazenil, on the arrhythmogenic serum concentration of epinephrine (ACE) in six halothane-anesthetized dogs. Midazolam was administered as a loading dose (1.5 mg/kg over 5 min) followed by a 4.5-h infusion at two rates (10 and 40 micrograms.kg-1 x min-1) to achieve and maintain predetermined clinical and supraclinical plasma midazolam concentrations. The arrhythmogenic serum concentration of epinephrine determinations were made prior to midazolam infusion, following 2 h of midazolam infusion and following 3.5 h of midazolam infusion and 1 mg flumazenil/kg i.v. Saline control studies were also performed in four of the six dogs. RESULTS: Plasma midazolam concentrations ranged from 363 to 855 ng/ml in the low-dose infusion study, essentially spanning the clinically effective range for humans. In the high-dose infusion study, plasma midazolam concentrations were up to four times greater, ranging from 1168 to 3563 ng/ml. The arrhythmogenic serum concentration of epinephrine values were unchanged following low-dose midazolam infusion and saline. In the high-dose study, ACE increased from baseline values of 68 +/- 13 (SEM) ng/ml to 112 +/- 25 ng/ml (P = .03) following midazolam infusion and decreased to 79 +/- 13 ng/ml with flumazenil administration. Plasma midazolam concentrations, however, were poorly correlated with ACE values normalized for control ACE (ACE ratio). Diastolic arterial pressure was significantly depressed following both low-dose (-14%) and high-dose (-19%) midazolam infusion. This decrease in blood pressure was unaffected by flumazenil administration. Other hemodynamic parameters were unaffected by drug treatment. CONCLUSIONS: This study has demonstrated that midazolam infusion results in either no effect (with clinical plasma midazolam concentrations) or flumazenil-reversible suppression (with supraclinical concentrations) of halothane-epinephrine arrhythmogenesis. PMID- 8424550 TI - Effects of ketamine on cardiovascular responses mediated by N-methyl-D-aspartate receptor in the rat nucleus tractus solitarius. AB - BACKGROUND: Ketamine is often administered to patients in whom hemodynamic instability is suspected. This study was designed to investigate the effect and possible site of action of ketamine on baroreflex response in rats. The site focused upon was the N-methyl-D-aspartate (NMDA) receptor of the nucleus tractus solitarius (NTS). METHODS: The effect of ketamine upon the baroreflex response was examined in urethan-anesthetized rats. The baroreflex was elicited by traction of the left carotid artery. Additional studies tested the acute hypotension and effect of ketamine on the hemodynamic response elicited by microinjection of NMDA into the NTS. RESULTS: Upon traction, the mean arterial pressure (MAP) decreased by 28.0 +/- 1.0 (mean +/- SE) mmHg and the heart rate (HR) decreased by 7.5 +/- 0.8 beats/min. Ketamine (9 mg/kg i.v.) attenuated these responses with a resultant decrease in MAP and HR of 3.6 +/- 1.3 mmHg and 1.6 +/- 0.2 beats/min, respectively (P < .01). It also suppressed the NMDA-induced decrease in MAP from 47 +/- 5 to 6 +/- 0.7 mmHg and delayed the decrease in HR. D 2-amino-5-phosphonovalerate, a competitive NMDA antagonist, blocked the NMDA induced decrease in MAP from 47 +/- 5 to 18 +/- 6 mmHg and delayed the decrease in HR. CONCLUSIONS: These findings support the view that ketamine might attenuate cardiovascular responses such as baroreflex by interacting, at least in part, with the NMDA receptor in the NTS. PMID- 8424551 TI - Fiber-type caffeine sensitivities in skinned muscle fibers from humans susceptible to malignant hyperthermia. AB - BACKGROUND: The response to contracture tests may depend upon the relative proportion of muscle fiber types within the muscle specimen. To determine whether a difference in fiber-type caffeine sensitivities exists between malignant hyperthermia susceptible (MHS) and malignant hyperthermia-nonsusceptible (MHN) skeletal muscle, we compared the fiber-type caffeine sensitivities in chemically skinned muscle fibers dissected from vastus lateralis muscle from 15 MHS and 16 MHN patients. METHODS: Muscle fiber type was determined in each fiber by the difference in strontium-induced tension measurements and in 36 fibers, after contracture testing, by ATPase enzyme histochemistry. Caffeine sensitivity was defined as the threshold concentration inducing more than 10% of the maximal tension obtained with a calcium 1.6 x 10(-2) mM solution. RESULTS: Significant difference in the mean (+/- SD) caffeine sensitivity was found between type I MHS fibers (2.63 +/- 0.85 mM) versus type II MHS fibers (3.47 +/- 1.2 mM) and between type I MHN fibers (5.89 +/- 1.8 mM) versus type II MHN fibers (10.46 +/- 2.6 mM). The mean (+/- SD) caffeine sensitivities for a given muscle fiber type (I or II) were different between groups of MHS and MHN patients. Both type I and II MHS fibers had significantly lower caffeine sensitivities, and this increase in caffeine sensitivity was significantly smaller in type I than in type II fiber. CONCLUSIONS: The current study indicates that a truly MHS patient cannot have a false-negative result solely related to abnormal type II fibers contained in a given muscle strip. Although the occurrence of a very high proportion of type I fibers in MHN human muscle could result in a false-positive contracture outcome, such an occurrence is expected to be rare. PMID- 8424552 TI - A reliable accounting system for controlled substances in the operating room. AB - BACKGROUND: Drug abuse is a leading occupational hazard for operating room personnel. Easy access to controlled substances allows drug dependence to develop and flourish. A system that accurately audits the distribution of controlled substances used in the operating room may decrease the onset of drug abuse and make it easier to identify drug addicts. A simple, inexpensive, and reliable system that improves accountability of controlled substances is described in detail. METHODS: This system involves participation by anesthesiologists, operating room nurses, and pharmacists to accurately record amount and type of drugs dispensed, used, wasted, and returned. Periodic, random, qualitative, and quantitative analyses of drugs returned for wastage are performed. RESULTS: In the first 6 months in which the system was used, 6,336 patients were treated and 7,182 ampules of controlled substances were dispensed. Thirty-seven incident reports describing deviations from the protocol occurred. In each case an explanation for the discrepancy was determined and compliance with the protocol was subsequently improved. No cases of drug diversion were discovered or suspected. CONCLUSIONS: The system described is simple and inexpensive to implement and has improved accountability for controlled substance management. PMID- 8424553 TI - The diagnosis and management of a perianesthetic cerebral aneurysmal rupture aided with transcranial Doppler ultrasonography. PMID- 8424554 TI - Management of a patient with type IIC von Willebrand's disease during coronary artery bypass graft surgery. PMID- 8424555 TI - Postdural puncture headache after fluoroscopically guided lumbar paravertebral sympathetic block. PMID- 8424556 TI - Are new drugs cost-effective for patients undergoing ambulatory surgery? PMID- 8424557 TI - Inflammatory cutaneous reactions to epidural catheters. PMID- 8424558 TI - Spinal epidural abscess: three cases following spinal epidural injection demonstrated with magnetic resonance imaging. PMID- 8424559 TI - Electroencephalographic monitoring during cardiac surgery. PMID- 8424560 TI - Quantitative electroencephalographic monitoring during cardiopulmonary bypass. PMID- 8424562 TI - Potential fresh gas flow leak through Drager Vapor 19.1 vaporizer with key-index fill port. PMID- 8424561 TI - Comparison of ondansetron versus placebo to prevent postoperative nausea and vomiting in women undergoing ambulatory gynecologic surgery. AB - BACKGROUND: Postoperative nausea and emesis, especially in ambulatory surgical patients, remains a troublesome problem. This study was performed to compare the incidence of nausea and emesis during the 24-h postoperative period in ondansetron-treated patients versus placebo-treated patients. METHODS: Using a randomized prospective double-blind study design, women between the ages of 18 and 70 yr undergoing gynecologic surgical procedures with general opioid anesthesia on an outpatient basis were enrolled. Ondansetron or placebo was administered prior to induction of anesthesia. Patients were stratified according to history of nausea and emesis during previous exposure to general anesthesia and randomized to dose received. RESULTS: Data from the 544 women showed that all doses of intravenous ondansetron tested (1, 4, and 8 mg) were significantly more effective (62%, 76%, and 77%, respectively) than placebo (46%) in reducing the incidence of emesis following surgery until 24 h after recovery room entry. All these doses were more effective than placebo in patients with no prior history of emesis following surgery and the 4- and 8-mg doses were more effective than placebo in patients with a prior history of emesis following surgery. All doses of ondansetron tested were generally well tolerated with adverse events, clinical laboratory tests, and recovery room vital signs similar to those of placebo. Serum aspartate transaminase (AST) was increased in five patients (1 mg, 2 patients; 4 mg, 1 patient; 8 mg, 2 patients). In the three patients in whom subsequent analysis were performed, the serum AST had decreased to preoperative levels. CONCLUSIONS: Ondansetron given intravenously to prevent postoperative nausea and emesis was highly effective in the 4- and 8-mg doses in women having ambulatory gynecologic surgery. PMID- 8424563 TI - Pulmonary hypertension and liver transplantation. PMID- 8424564 TI - Cauda equina syndrome and continuous spinal anesthesia. PMID- 8424566 TI - Undetected leak in corrugated circuit tubing in compressed configuration. PMID- 8424565 TI - Regulation of skeletal muscle acetylcholine receptors. PMID- 8424567 TI - Preemptive analgesia or anoci-association. PMID- 8424568 TI - Intraoperative somatosensory evoked potential monitoring predicts peripheral nerve injury during cardiac surgery. AB - BACKGROUND: Brachial plexus injury may occur without obvious cause in patients undergoing cardiac surgery. To determine whether such peripheral nerve injury can be predicted intraoperatively, we monitored somatosensory evoked potentials (SEPs) from bilateral median and ulnar nerves in 30 patients undergoing coronary artery bypass surgery. METHODS: SEPs were analyzed for changes during central venous cannulation and during use of the Favoloro and Canadian self-retaining sternal retractors, events hereto implicated in brachial plexus injury. Brachial plexus injury was evaluated during physical examination in the postoperative period by an individual blinded to results of SEP monitoring. RESULTS: Central venous cannulation was associated with transient changes in SEPs in four patients (13%). These changes occurred intermittently during insertion of the cannula but completely resolved within 5 min. Postoperative neurologic deficits did not occur in these cases. Use of the Canadian and Favoloro retractors was associated with significant changes in 21 patients (70%). In 16 of these, waveforms reverted toward baseline levels intraoperatively and were not associated with postoperative neurologic deficits. Five patients demonstrated a neurologic deficit postoperatively. In each of these, SEP change associated with use of surgical retractors persisted to the end of surgery compared to the immediate pre bypass period. CONCLUSION: Intraoperative upper extremity SEPs may be used to predict peripheral nerve injury occurring during cardiac surgery. PMID- 8424569 TI - Transdermal fentanyl for cancer pain. Repeated dose pharmacokinetics. AB - BACKGROUND: The transdermal therapeutic system (fentanyl), or TTS(fentanyl), continuously delivers fentanyl for up to 72 h. The transdermal therapeutic system (fentanyl)-100 delivers approximately 100 micrograms/h. The repeated dose pharmacokinetics of this drug using the recommended dosing interval have not been evaluated previously and were determined in the present study. METHODS: Blood samples were obtained from ten opioid-tolerant cancer patients who received five applications of TTS(fentanyl) at 72-h intervals. A sample of venous blood was taken before each dose; multiple samples were taken during and after the fifth application. A gas chromatographic/mass spectrometry method was used to assay fentanyl (limit of detection 0.2 ng/ml). RESULTS: For the fifth dose, the mean (SD) maximum concentration was 2.6 (1.3) ng/ml and the mean (SD) area under the serum fentanyl concentration-time curve (0-72 h) was 116.9 (59.9). Following removal of the system, the mean (SD) apparent half-life was 21.9 (8.9) h. There were no differences among the serum fentanyl concentrations measured before the second through fifth doses. Fentanyl absorption was 47% complete at 24 h, 88% complete at 48 h, and 94% complete at 72 h. The mean (SD) dose delivered during the 72-h period was 4.3 (1.1) mg. A first-dose trough concentration predicted from fifth-dose kinetics and the actual first-dose trough concentration were very similar. Adverse effects ascribed to the transdermal system were minimal. CONCLUSIONS: These results suggest that steady-state serum concentrations are approached by the second dose of TTS(fentanyl) and that the kinetics are stable with repeated dosing. The apparent half-life following system removal is relatively long, indicating ongoing absorption from a subcutaneous depot. PMID- 8424570 TI - Anesthesia for thoracoscopic laser ablation of bullous emphysema. AB - BACKGROUND: We describe the anesthetic management for a new surgical procedure: laser ablation of emphysematous bullae via thoracoscope. Although thoracoscopy is not new, this is the first description of a series of patients with bilateral, chronic lung disease who underwent long periods of one-lung ventilation (OLV) during thoracoscopic therapy. METHODS: Twenty-six laser ablation procedures were performed in 22 patients. The patients were elderly (mean age 63 yr) with a large incidence of coexisting cardiovascular disease. Most required chronic home oxygen therapy. Patients were monitored invasively, and hemodynamic data were recorded every 5 min. Arterial blood gas analyses were performed every 15 min. Comparisons were made between three intraoperative periods: two-lung ventilation (TLV) before thoracoscopy, OLV during thoracoscopy, and TLV after thoracoscopy. RESULTS: All patients survived the operation despite a mean OLV duration of 170 min, but several experienced serious intraoperative problems, such as hypoxemia or hypotension. Hypoxemia was treated with nondependent lung continuous positive airway pressure and dependent lung positive end-expiratory pressure. In all patients the lungs were adequately ventilated, but bronchopleural fistulae occurred upon return to TLV in every case. The resulting air leaks, often 50% of inspired tidal volume, required the use of a pressure-cycled ventilator to maintain oxygenation. Postoperative air leaks greater than 50% of inspired tidal volume usually required subsequent surgical correction, while smaller leaks resolved spontaneously. Mechanical ventilation was required for an average of 5 days. Eighty-four percent have survived at least 6 months, and nearly all survivors report symptomatic improvement. CONCLUSIONS: Ablation of bullae appears to provide symptomatic improvement, and thoracoscopy might be better tolerated than thoracotomy, especially in patients with severe bullous emphysema. PMID- 8424571 TI - Midazolam enhances anterograde but not retrograde amnesia in pediatric patients. AB - BACKGROUND: Midazolam sedation has been shown to diminish recall of one to four cards shown prior to induction of general anesthesia in pediatric patients. This promising but limited finding prompted us to investigate the effect of midazolam sedation on retrograde and anterograde recall and recognition in children scheduled for elective surgery. METHODS: Forty patients aged 4-10 yr were randomized using a double-blind study design to receive either 0.2 mg/kg intranasal midazolam or 0.2 ml/5 kg placebo (distilled water) using a Devilbiss #286 atomizer. To assess postoperative memory of preoperative events, recall and recognition tasks were performed using a series of picture cards designed for this purpose. Retrograde amnesia was measured by postoperative recall and recognition of cards shown prior to midazolam/placebo administration, and anterograde amnesia was measured by postoperative recall and recognition of cards shown during the interval between midazolam/placebo administration and induction of general anesthesia. RESULTS: Compared to placebo, the midazolam group experienced a significant postoperative reduction in ability to both recall (P < .003) and recognize (P < .001) cards shown subsequent to midazolam/placebo administration (anterograde amnesia). In distinction, there was no difference between groups in postoperative ability to recall or recognize cards shown prior to midazolam/placebo administration (retrograde amnesia). CONCLUSIONS: These results support and extend the inference that midazolam diminishes anterograde recall. In addition, our findings indicate that midazolam diminishes anterograde recognition, thereby providing partial anterograde amnesia without affecting retrograde memory in pediatric patients. PMID- 8424572 TI - Clinical significance of pulmonary aspiration during the perioperative period. AB - BACKGROUND: Pulmonary aspiration of gastric contents during the perioperative period may be associated with postoperative mortality or pulmonary morbidity. Recent determination of the incidence of perioperative pulmonary aspiration and evaluation of factors related to clinical outcomes is lacking. METHODS: We retrospectively reviewed the perioperative courses of 172,334 consecutive patients 18 yr of age or older who underwent 215,488 general anesthetics for procedures performed in all surgical specialties from July 1985 to June 1991. Pulmonary aspiration was defined as either the presence of bilious secretions or particulate matter in the tracheobronchial tree or, in patients who did not have their tracheobronchial airways directly examined after regurgitation, the presence of an infiltrate on postoperative chest roentgenogram that was not identified by preoperative roentgenogram or physical examination. RESULTS: Pulmonary aspiration occurred in 67 patients (1:3,216 anesthetics). Fifteen aspirations occurred in 13,427 (1:895) anesthetics of patients undergoing emergency surgery, and 52 occurred in 202,061 (1:3,886) anesthetics of patients undergoing elective surgery (P < .001). Of the 66 patients who survived their surgery, 42 (64%) did not develop a cough or wheeze, a decrease in arterial hemoglobin oxygen saturation while breathing room air > 10% less than the preoperative value, or radiographic abnormalities within 2 h of aspiration. These 42 patients had no respiratory sequelae. Of the 24 patients who had one or more of these findings, 13 required mechanical ventilatory support for more than 6 h. Three of the six patients whose lungs required mechanical ventilation for more than 24 h died from pulmonary insufficiency (overall mortality = 1:71,829 anesthetics). CONCLUSIONS: This study suggests that patients with clinically apparent aspiration who do not develop symptoms within 2 h are unlikely to have respiratory sequelae. PMID- 8424573 TI - A randomized, double-blind evaluation of ketorolac tromethamine for postoperative analgesia in ambulatory surgery patients. AB - BACKGROUND: Given the trend toward early discharge of patients after surgery and the inherent adverse effects of opioid analgesics, we compared a new nonsteroidal antiinflammatory drug, ketorolac tromethamine, given intravenously (iv) and then orally, with two commonly prescribed opioid analgesics in ambulatory patients for up to 1 week after surgery. METHODS: In this study incorporating a double-blind, multi-dose design, 221 patients who had moderate or severe pain after surgery were randomized to one of three treatment groups: group K30 received 30 mg iv ketorolac twice, then 10 mg iv every 30 min as required to control pain, up to six doses, followed by 10 mg oral ketorolac every 4-6 h; group F50 received 50 micrograms iv fentanyl at the same time intervals as in group K30, followed by 60 mg codeine plus 600 mg acetaminophen (C+A) orally every 4-6 h; and group F10 received the same combination as did group F50, but only 10 micrograms fentanyl per dose. RESULTS: Compared with 50 micrograms fentanyl iv, 30 mg iv ketorolac provided delayed but otherwise equivalent analgesic effects and was associated with similar side effects. Compared with C+A, 10 mg oral ketorolac was associated with a lower incidence of nausea and somnolence and earlier return of bowel function but not better pain relief, drug tolerability, quality of life, or psychologic well-being. CONCLUSIONS: Ketorolac, when used in an iv and then oral sequence, is a safe and effective analgesic in the ambulatory surgery setting. It has a slower onset than fentanyl, but causes fewer side effects than C+A. PMID- 8424574 TI - Effects of thoracic extradural block on diaphragmatic electrical activity and contractility after upper abdominal surgery. AB - BACKGROUND: Upper abdominal surgery (UAS) induces diaphragmatic dysfunction. Thoracic extradural block (TEB) using 0.5% bupivacaine improves some pressure and motion indices of diaphragmatic function. However, no direct information on diaphragmatic activity is available after UAS. The aim of this study was to assess diaphragmatic electrical activity (Edi) after UAS before and after TEB. METHODS: A postoperative electromyogram was obtained, using intramuscular electrodes inserted by the surgeon in the costal and crural parts of the diaphragm, in 14 patients undergoing abdominal aortic surgery. Tidal changes in abdominal (VAB) and rib-cage (VRC) volumes, and gastric (delta Pgas), esophageal (delta Pes), and transdiaphragmatic (delta Pdi) pressures were used to measure tidal volume (VT) and respiratory rate and to provide indirect indices of diaphragmatic activity from the two ratios VAB/VT and delta Pgas/delta Pdi. These respiratory variables were obtained preoperatively. Postoperatively, measurements including Edi were obtained before and after a segmental epidural block, reaching a T4 level was achieved with 0.5% plain bupivacaine. RESULTS: Upper abdominal surgery induced an increase in respiratory rate (+28 +/- 15%; P < .01), associated with a decrease in VAB/VT (from 0.75 +/- 0.11 to 0.07 +/- 0.08; P < .01), delta Pgas/delta Pdi (from 0.3 +/- 0.08 to 0.01 +/- 0.19; P < .05), and VT (-30 +/- 14%; P < .01). After surgery, all patients exhibited electrical diaphragmatic activity that increased with TEB by 48 +/- 28% (P < .01) and 60 +/- 22% (P < .001) for the cural and costal segments, respectively. The ratio delta Pdi/Edi, used to evaluate diaphragmatic contractility, was not modified by TEB. Tidal volume, respiratory rate, and delta Pgas/delta Pdi returned to preoperative levels, whereas VAB/VT increased but remained different from preoperative values. CONCLUSIONS: This study demonstrates that TEB produces an increase in diaphragmatic activity, identical for the two segments of the muscle. Interruption of afferents that produce an inhibitory effect on diaphragmatic activity appears the most attractive hypothesis to explain the consequences of TEB after UAS. PMID- 8424575 TI - Halothane decreases the rate of production of cerebrospinal fluid. Possible role of vasopressin V1 receptors. AB - BACKGROUND: Circulating vasoactive hormones (e.g., vasopressin) play an important role in the regulation of blood flow to the choroid plexus and the rate of cerebrospinal fluid (CSF) production. We tested the hypothesis that halothane decreases CSF production through a vasopressin-related mechanism and examined the related changes in blood flow to the choroid plexus. METHODS: Using ventriculocisternal perfusion, CSF production was measured in chloralose anesthetized, normothermic rabbits whose lungs were mechanically ventilated. Rabbits received either 0.5 minimum alveolar concentration (MAC; end-tidal) of halothane (added to a preestablished chloralose anesthetic), 0.5 MAC of halothane in the presence of a vasopressin V1 antagonist (iv), or the V1 antagonist alone. In addition, we examined animals in which no intervention was made (time control) and animals subjected to a 25% decrease in mean blood pressure produced by hemorrhage, with and without the V1-antagonist. In a separate series of rabbits, regional and total blood flows to the brain and the choroid plexus were measured using radioactive microspheres. These studies were carried out under similar conditions, except that the effects of end-tidal 0.25, 0.5, and 1 MAC of halothane were examined in each rabbit (each added to a preestablished chloralose anesthetic). RESULTS: Under control conditions, blood flow to the choroid plexus averaged 351 +/- 198 ml.min-1.100 g-1 (mean +/- SD) and CSF production averaged 10.1 +/- 1.9 microliters.min.-1. Halothane (0.25, 0.5, and 1 MAC) did not alter choroid plexus blood flow but decreased CSF production by 28 +/- 6% at 0.5 MAC (P < .05). In contrast, 1 MAC of halothane increased total blood flow to the brain by 20 +/- 25% (P < .05). The V1 antagonist, which did not affect production of CSF when given alone, prevented the decrease in CSF production in response to halothane. Hemorrhage decreased blood flow to the choroid plexus but not to the brain, and the V1 antagonist attenuated the decrease in the rate of CSF production by hemorrhage (34 +/- 11% vs. 48 +/- 18%, P < .05). CONCLUSIONS: Halothane decreases CSF production with no net change in the blood flow to the choroid plexus. Decrease in CSF production appears to be mediated through a vasopressin-related mechanism and not to the blood pressure decrease seen during halothane anesthesia. PMID- 8424576 TI - Effects of etomidate on the cardiac papillary muscle of normal hamsters and those with cardiomyopathy. AB - BACKGROUND: Etomidate has been shown to induce no significant inotropic effect on normal myocardium, but its effects on diseased myocardium remain unknown. METHODS: The effects of etomidate (1 and 5 micrograms/ml) on the intrinsic contractility of left ventricular papillary muscle from normal hamsters and those with cardiomyopathy (strain BIO 82.62, 6 months old) were investigated in vitro (Krebs-Henseleit solution, 29 degrees C, pH 7.40, Ca++ 2.5 mM, stimulation frequency 3/min). RESULTS: The contractility of papillary muscles from hamsters with cardiomyopathy was less than that of controls, as shown by the decrease in maximum shortening velocity (-25%, P < .001), isometric active force (-45%, P < .01), peak power output (-57%, P < .01), and sarcoplasmic reticulum function (P < .01). Etomidate did not induce a significant inotropic effect, as shown by the absence of changes in maximum shortening velocity and active isometric force, except at 5 micrograms/ml in cardiomyopathic hamsters (+8 +/- 10%, P < .05). The effects of etomidate on these inotropic parameters were not different in normal and cardiomyopathic hamsters. Etomidate impaired contraction-relaxation coupling under low load in both groups, suggesting that etomidate decreased sarcoplasmic function. This impairment was less (P < .02) pronounced in cardiomyopathic muscles. The effects of etomidate on contraction-relaxation coupling under heavy load were not different between groups. In both groups, etomidate had no effect on the peak power output and the curvature of the total force-velocity curve, suggesting that it did not modify the muscle myothermal economy. CONCLUSIONS: Etomidate had only a slight effect on the intrinsic mechanical properties of hamster cardiac papillary muscles, and these effects did not depend on the pathophysiologic state of the myocardium. These results may be clinically useful as, unlike etomidate, most anesthetics depress myocardial contractility. PMID- 8424577 TI - Hypocapnia worsens arterial blood oxygenation and increases VA/Q heterogeneity in canine pulmonary edema. AB - BACKGROUND: Hyperventilation frequently is employed to reduce carbon dioxide partial pressure in patients in the operating room and intensive care unit. However the effect of hypocapnia on oxygenation is complex and may result in worsening in patients with preexisting intrapulmonary shunt. To better define the interplay between hypocapnia and oxygenation, the effects of hypocapnia and hypercapnia on the matching of ventilation (VA) and perfusion (Q) were studied in dogs with oleic acid-induced pulmonary edema, using the multiple inert gas elimination technique. METHODS: Eight pentobarbital-anesthetized, closed-chested dogs were administered 0.06 ml/kg of oleic acid at least 150 min prior to study. Ventilation was set with an FIO2 of 0.90, a tidal volume of 20 ml/kg, and a respiratory rate of 35 breaths/min. The arterial carbon dioxide tension (PaCO2) was varied in a randomized order to three levels (26, 38, and 50 mmHg) by altering the amount of CO2 in the inspired gas mixture. Gas exchange was assessed by true shunt, dead space, the log standard deviation of the perfusion (log SDQ) and the ventilation (log SDV) distributions, and the tracer inert gas arterial alveolar difference ([a-A]D) area. RESULTS: Cardiac output (4.1 +/- 0.4 L/min), mean pulmonary artery pressure (25 +/- 1 mmHg), inert gas shunt (22 +/- 3%), and dead space (38 +/- 4%) during normocapnia were not different from that during hypocapnia and hypercapnia. Hypocapnia increased (P < .05) the alveolar-arterial oxygen tension difference (P[A-a]O2) and decreased (P < .05) the arterial blood oxygen tension (PaO2, 181 +/- 33 mmHg vs. 221 +/- 35 mmHg with normocapnia). P[A a]O2 and PaO2 were unaffected by hypercapnia. During hypocapnia, VA/Q inequality increased, demonstrated by an increase (P < .05) in log SDQ (1.60 +/- 0.15 vs. 1.35 +/- 0.19 with normocapnia) and in the [a-A]D area (0.63 +/- 0.09 vs. 0.50 +/ 0.09 with normocapnia) indexes of VA/Q heterogeneity. During hypercapnia, the [a A]D area (0.63 +/- 0.11) and log SDV (1.13 +/- 0.10 compared to 0.97 +/- 0.12 with normocapnia) also were increased (P < .05). With hypocapnia, there was a small but insignificant increase in blood flow to shunt and low VA/Q areas (29 +/ 4% compared to 26 +/- 4% with normocapnia). In the presence of a high FIO2, this small increase in shunt and low VA/Q may result in a significant decrease in PaO2. CONCLUSIONS: Both hypocapnia and hypercapnia were associated with an increased VA/Q inequality. However, PaO2 decreased and P[A-a]O2 increased with only hypocapnia. These results suggest that hyperventilation to reduce PaCO2 may be detrimental to arterial PO2 in some patients with lung disease. PMID- 8424578 TI - Functional benefits of peripheral vascular bypass surgery for patients with intermittent claudication. AB - Many patients with peripheral arterial disease (PAD) undergo peripheral bypass surgery to relieve the symptom of intermittent claudication. However, measurement of graft patency alone, assessed by change in ankle blood pressure, may not adequately reflect the improvement in functional status following the operation. Fourteen patients with moderately severe intermittent claudication were evaluated before and after bypass surgery to asses changes in hemodynamics (by resting ankle-brachial indices-ABIs), exercise performance (by a graded treadmill protocol), and community-based walking ability (by a questionnaire that characterizes self-reported walking speed and distance). Six weeks after surgery, resting ABIs improved in surgical patients (from 0.56 +/- 0.09 to 0.93 +/- 0.16, P < 0.05). Peak treadmill walking time improved from 6.2 +/- 5.9 to 11.8 +/- 7.1 min (90%), peak oxygen consumption from 15.5 +/- 6.5 to 19.1 +/- 8.5 mL/kg/min (23%), and pain-free walking time from 1.5 +/- 0.4 to 6.0 5.5 min (290%). Questionnaire scores for walking distance improved by 203% and for walking speed by 130% (all P < 0.05). After twelve weeks, improvements other than pain-free walking time were maintained. Changes in peak treadmill performance or questionnaire scores were not correlated with the initial ABI, changes in ABI six or twelve weeks after surgery, or the type or extent of bypass surgery. The results demonstrate that surgical treatment of PAD, indicated for the relief of intermittent claudication, improves exercise performance and self-reported community-based walking ability. The change in functional status of the patient is an important outcome of surgery that cannot be predicted from routine noninvasive testing alone and should be measured directly. PMID- 8424579 TI - Predictive value of manual ECG-monitored exercise test before abdominal aortic or peripheral vascular surgery. AB - The value of manual ECG-monitored exercise test was studied in 58 patients (39 men and 19 women) with intermittent claudication scheduled for abdominal or peripheral vascular surgery. There was a high incidence of well-known risk factors (hypercholesterolemia, smoking, and hypertension) in men and women. History of coronary artery disease (CAD) was present in 29.3% of the patients. No correlation was found between patients with single or multiple stenoses in lower extremity arteries (angiography) and CAD. Ischemic response (ECG) to manual exercise testing was positive in 14 patients (24.1%), of whom 2 (14.3%) had major cardiovascular postoperative complications. Both died of acute myocardial infarction within fourteen days. They had slow recovery (thirty to sixty minutes) of the ST segment (0.3-0.6 mV). The authors conclude that manual exercise testing might reveal serious CAD in patients with severe occlusive peripheral arterial disease of the lower extremities. In cases with slow recovery of ischemic response to exercise, coronary bypass or angioplasty could be advantageous before major vascular surgery. PMID- 8424580 TI - Beneficial effects of intermittent suction and pressure treatment in intermittent claudication. AB - The present study reports on the effects of a physical treatment modality in patients with intermittent claudication. During this treatment a major part of the skin surface is subjected to intermittent suction and pressure. In a previous, preliminary study the authors found a beneficial effect of this treatment in intermittent claudication. The study included 34 patients with moderate, stable intermittent claudication. Twenty-two patients participated in a double-blinded, randomized trial comparing the effects of 25 treatments to 25 placebo applications given over a period of two months. Twelve patients participated in an open trial investigating the possible effects of the treatment on platelet aggregation and fibrinolysis. Pain-free and maximal walking distances were measured on a treadmill, and systolic blood pressure was measured on the upper limb, the ankle, and the first toe bilaterally. The threshold for adenosine diphosphate (ADP)-induced platelet aggregation was tested, and the fibrinolytic activity was estimated from the euglobulin clot lysis time. Active treatment resulted in significant improvements in pain-free and maximal walking distances, whereas no changes could be found during placebo administration. The treatment caused significant increments in the ADP thresholds for platelet aggregation, while the effects on fibrinolysis were uncertain. It is concluded that intermittent suction and pressure treatment offers a new approach for conservative treatment of intermittent claudication. PMID- 8424581 TI - Clinical results of epidural spinal cord electrical stimulation in patients affected with limb-threatening chronic arterial obstructive disease. AB - Between 1982 and 1990, 76 patients (33 women, 43 men, mean age 71.4 +/- 10 years) affected with limb-threatening peripheral vascular disease (claudication < 20 m: 3 patients; rest pain: 10 patients; necrosis1 smaller than 3 cm2: 28 patients; necrosis2 larger than 3 cm2: 35 patients) not amenable to medical and/or surgical therapy, were treated by epidural spinal cord electrical stimulation (ESES). Effectiveness of ESES was evaluated by consideration of pain control, walking distance, and healing of ischemic lesions. At a mean follow-up of twenty-six months (range: one to seventy-six) 44 limbs (58%) were amputated (rest pain 2; necrosis1 13; necrosis2 29) and 39% of necrotic lesions smaller than 3 cm2 healed. The overall limb salvage rate was 42%. Pain control was obtained in 80% of patients at the one-year and 75% at the two-year follow-up, with infrequent use of pain relievers. Despite the poor clinical results observed, the limb salvage rate testifies to the effectiveness of ESES in limb-threatening ischemia. Moreover, the authors noticed a good ESES effect on pain relief, maximal in the early and intermediate postimplant periods. In conclusion ESES must be considered the last resort in peripheral vascular disease in patients in whom medical and/or surgical therapies are ineffective or impossible. Necrotic lesions larger than 3 cm2 contraindicate, in their opinion, ESES implant. PMID- 8424582 TI - Duplex ultrasonography for the detection of deep vein thrombi after total hip or knee arthroplasty. AB - The usefulness of real-time duplex ultrasonography (DU) as a screening test for deep vein thrombosis (DVT) in high-risk patients remains uncertain. To determine the sensitivity and specificity of DU for the detection of DVT, the authors prospectively studied 178 consecutive patients after total hip (n = 113) or total knee (n = 66) arthroplasty. The deep veins from the inguinal ligament to the ankle were examined first by continuous wave and then by pulsed Doppler signals as needed with real-time gray-scale ultrasound imaging using the criteria of vein noncompressibility to define DVT. Ascending contrast venography was performed within twelve hours after DU studies. Venograms and DU were interpreted independently. DU was attempted on 177 lower extremities (2 patients refused) but was judged adequate for interpretation for only 145 (82%). Venography could not be performed for 28 lower extremities and was technically inadequate for 8 studies. The primary analysis included 119 examinations for which adequate DU and ascending venograms were interpreted. DU was positive in 17 of 27 lower extremities with DVT (23 calf, 4 proximal) diagnosed by venography (sensitivity = .63; 95% confidence interval [CI] = .42 to .81), and DU was negative in 85 of 92 lower extremities with normal venograms (specificity = .92; 95% CI = .85 to .97). A secondary analysis of 81 prospectively collected anatomically complete DU studies demonstrated a sensitivity of .80 (95% CI = .56 to .94) and a specificity of .90 (95% CI = .80 to .96).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424583 TI - Incidence of carotid artery atherosclerosis in patients with coronary artery disease. AB - The natural history of coronary artery disease (CAD) is often complicated by cerebrovascular accidents. The real incidence of atherosclerotic lesions of carotid arteries in coronary patients is not well established. In order to detect a high-risk group for stroke development, 184 patients affected by CAD, either partially symptomatic or asymptomatic for carotid artery stenosis, underwent Echo Doppler ultrasonography (duplex scanning) of supra-aortic branches. Significant carotid stenosis (> 50%) was demonstrated in 51 subjects (27.7%); 21 subjects (41.2%) were partially symptomatic (dizziness, vertigo, lipothymia, etc), and 30 subjects (58.8%) were completely asymptomatic. The authors' data suggest that carotid disease can develop concurrently with coronary disease in a significant proportion of patients, even though completely asymptomatic. In order to obtain optimal long-term results, both coronary and carotid artery disease require appropriate evaluation and either medical or surgical management. For these reasons they recommend duplex scanning as a routine screening procedure in patients affected by CAD. PMID- 8424584 TI - Decreased platelet activity without change in fibrinolytic activity after low dosages of fish oil. AB - The effects of low-dose (350 mg daily) polyunsaturated fatty acids from fish oil, either in gelatine capsules or microencapsulated, were investigated in a non blind, randomized, crossover study of 12 healthy male volunteers. The authors measured the incorporation of eicosapentaenoic acid (EPA) into platelets membranes, platelet aggregability by adenosine diphosphate (ADP) and adrenaline, and fibrinolytic activity as euglobulin lysis time, after two and six weeks of therapy. Both formulations resulted in increased incorporation of EPA into platelet membranes, and the microencapsulated formulation also significantly increased the platelet level of ADP for irreversible aggregation (by about 60%). Fish oil in gelatine capsules had a smaller, non-significant effect in the same direction. Both formulations reduced adrenaline-induced aggregability, but the effects did not attain significance. Neither formulation altered fibrinolysis. The data show that low doses of fish oil in microencapsulated form significantly lower platelet aggregability, without affecting fibrinolysis. PMID- 8424586 TI - Septal perforator artery angioplasty: the advantage of application of an ultralow profile balloon system--a case history. AB - Coronary artery disease can affect the septal perforator arteries and cause clinical sequelae, including angina, infarction of the interventricular septum, arrhythmias, and congestive heart failure. However, the size and the course of the septal perforator arteries significantly differ from those of the major epicardial coronary vessels. These anatomic features present certain technical challenges for angioplasty of the septal arteries. This case demonstrates that application of an ultralow-profile balloon-over-the-wire system can meet the specific requirements of septal artery angioplasty and achieve adequate results. PMID- 8424585 TI - Capillary filtration and ankle edema in patients with venous hypertension: effects of Daflon. AB - Capillary filtration (CF) was assessed in 43 patients with venous hypertension and in 10 normal subjects by means of strain gauge plethysmography. The patients were then divided into two treatment groups; 22 were treated with Daflon, 500 mg tid, and 21 with Daflon, 500 mg bid. The normal subjects were also treated (500 mg tid). After four weeks CF was significantly decreased in all subjects, the decrease being higher in the high-dose groups. CF did not significantly change in normal subjects. In conclusion Daflon decreases CF in subjects with venous hypertension and the CF decrease is dose related. PMID- 8424587 TI - Myocardial infarction complicating postsplenectomy thrombocytosis, with early left ventricular mural thrombus formation and cerebral embolism--a case report. AB - Thrombocytosis is a rare cause of ischemic cardiovascular and cerebrovascular events in patients with intrinsically normal coronary and cerebral vasculature. This report details the occurrence of inferior wall myocardial infarction (MI) consequent upon postsplenectomy thrombocytosis in a thirty-four-year-old man with angiographically normal coronary arteries. The MI was complicated by early left ventricular mural thrombus formation and embolic cerebral infarction. Combined anticoagulant and antiplatelet therapy was required to prevent the recurrence of ischemic events. PMID- 8424588 TI - Complete obstruction of right proximal pulmonary artery: a case report. AB - A fifty-five-year-old woman had worried about dyspnea for three years. A chest x ray appeared normal except for hypovascularity of the upper lung field. On angiograms, a wedge-shaped obstruction was observed at the left proximal pulmonary artery, and well-developed collateral circulation from the right thoracic artery to the internal costal arteries was observed. On the exercise test, exertional dyspnea developed with tachycardia and a decreased saturation of arterial oxygen. PMID- 8424589 TI - Compression of vein graft by hematoma during fibrinolytic therapy simulating intrinsic venous stricture--a case report. AB - Narrowing of a vein graft at an area of perigraft extravasation was encountered during fibrinolytic therapy for graft thrombosis. With cessation of fibrinolytic therapy, extravasation ceased and the vein lumen widened. Extrinsic compression and/or spasm due to a hematoma should be included as causes of vein graft narrowing in patients undergoing fibrinolytic therapy. PMID- 8424590 TI - Latex allergy. PMID- 8424591 TI - Chronic steroid-resistant urticaria. PMID- 8424592 TI - Isolation of Mycoplasma pneumoniae from asthmatic patients. AB - Asthmatic patients frequently develop wheezing after respiratory tract infection. Mycoplasma pneumoniae causes respiratory tract infections in children and in adults. The purpose of this study was to determine the frequency of recovery of M. pneumoniae from patients with asthma. Seventy-seven patients with asthma and 88 persons without asthma or any other respiratory tract disease (controls) were included in the study. Ages ranged from 8 months to 31 years. Throat swabs were taken and deposited in egg yolk broth with methylene blue in order to isolate M. pneumoniae. The bacterium was identified using inhibition of growth with homologous antiserum. The isolation rate in patients with asthma was 24.7% while that in controls was 5.7% (P < .01). The results suggest that M. pneumoniae colonizes a higher percentage of patients with asthma than controls and could possibly have induced the wheezing. PMID- 8424593 TI - Specific IgE antibodies in nasal secretions: correlation with serum values and clinical tests. AB - The presence of specific IgE in external secretions and the possibility of quantifying it has aroused the interest of many research workers. Nevertheless, there is still disagreement with regard to how useful this practice is from a clinical point of view. We have compared the presence of Dermatophagoides pteronyssinus (Dp) specific-IgE in the nasal secretion of 17 patients suffering from rhinitis with the results of prick tests and nasal and/or conjunctival challenge tests. As a preliminary step we have updated techniques and made a correlation between the parameters mentioned. The results for total IgE (n = 17) show a correlation coefficient (r = .39, NS) between serum values (251 +/- 239) U/L and those obtained in nasal secretion (28 +/- 36) U/L. (2) Serum and nasal-Dp specific-IgE values (13 +/- 15 and 5 +/- 9, respectively: (r = .72, P = .0011) match in 14 of the 17 determinations. In two of them, the serum values of Dp specific-IgE, but not the nasal ones, match the in vivo tests. In the others, it is the nasal secretion determination that matches the in vivo tests while the serum and nasal determinations match (82%). These results match in 86% of cases with prick tests and in 93% with nasal/conjunctival challenge tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424594 TI - Management of the patient with IgG subclass deficiency and/or selective antibody deficiency. PMID- 8424595 TI - Lung function and bronchial response to methacholine in young adults who had asthma in childhood. AB - Lung function and bronchial response to methacholine were studied in 47 young adults who had had childhood asthma, and after a period of 14-21 years, showed a different clinical evolution. At present, these subjects have been classified in four clinical groups: asymptomatic, rhinitic, asthmatic only due to exercise, and asthmatic. The same study was performed in 23 healthy individuals without personal histories of respiratory or allergic pathology. We found low spirometric basal values in both the asthmatic group (FEV1 and FEF25-75) and in the group with asthmatic responses to exercise (FEF25-75). No significant differences were found among asymptomatic, rhinitic, and control groups. While airway hyperreactivity was observed in patients who still had asthma, the bronchial response to methacholine in asymptomatic and rhinitic groups was not different from the control group. We conclude that both lung function and bronchial response to methacholine in most of the adults who had asthma in infancy and had been without asthmatic symptoms for many years are similar to those observed in the general population. PMID- 8424596 TI - Methodology in cardiac arrest research symposium. Introduction. PMID- 8424597 TI - Beyond methodology: improving clinical systems. PMID- 8424598 TI - Defining the benefits of rural emergency medical technician-defibrillation. AB - STUDY OBJECTIVE: To examine the success of rural emergency medical technician defibrillation (EMT-D) programs in relation to community population size. DESIGN AND SETTING: The four major studies addressing EMT-Ds in communities of fewer than 40,000 persons were reviewed. Additional, unpublished data on community size and neurologic condition of survivors were obtained from the authors. Survivors with prearrest neurologic status in communities with more than 15,000 persons were compared with those in communities of fewer than 15,000 persons. RESULTS: With the exception of the initial rural EMT-D study from Iowa, large communities had a higher rate of prearrest-level neurologic survival than small communities. Control communities appeared to have a slightly lower survival rate than small rural communities. CONCLUSION: Small rural communities cannot expect statistics for prearrest neurologic survival after cardiac arrest to be similar to those from large rural communities. Many emergency medical system factors, such as delay to system access, lack of first-responder CPR, and absence of full-time EMTs, may prevent EMT-D programs in small rural communities from experiencing comparable cardiac arrest survival. PMID- 8424599 TI - How much "significance" is significant? The transition from animal models to human trials in resuscitation research. AB - Animal models of resuscitation must be analyzed carefully before applying any interventions to human resuscitation. In addition to statistical concerns, the investigator must assess the clinical significance of the animal experience with an intervention. No single species is perfect, and the researcher must be aware of the major differences in primate, swine, canine, and rodent models. Interventions useful across species are more likely to be beneficial than those useful in one isolated species. After evaluating the animals used, the design in a series of animal experiments should be clinically applicable and the results reproducible. Finally, both process and outcome variables should be evaluated and improved in animal models before applying a new intervention to clinical care. PMID- 8424600 TI - In-hospital cardiac arrest. AB - Patients who suffer an in-hospital cardiac arrest represent a neglected and underutilized resource for resuscitation research. There exists an unwritten, but widely held, belief among resuscitation researchers that the in-hospital arrest population is unsuitable for resuscitation research because it is composed mostly of patients whose cardiac arrest is the terminal event of a fatal illness. Despite the large numbers of hospitalized patients on whom cardiac resuscitation is attempted each year, there are few reports and even less true research devoted to this clinical problem. This article, which is intended to be provocative, reviews and summarizes the existing literature on in-hospital resuscitation from cardiac arrest, considers the advantages of resuscitation research in this setting, and concludes with a challenge to resuscitation researchers. PMID- 8424601 TI - The ethics of cardiac arrest research. AB - In cardiac arrest research, prior informed consent is not available to resolve the conflict between the rights and well-being of subjects and the possible benefit to future patients. The right to autonomy is the fundamental right that is protected by the legal doctrine of informed consent. As a fundamental right, it cannot be balanced against other goods. Rather, it is a constraint, or trump, on the balancing of goods and can be overridden only for a narrow range of reasons: its recognition in a given case conflicts with another basic right, infringing the right will prevent great harm to others, and excluding a particular case from its scope will recognize and advance the right in the long run. Proxy consent, deferred consent, and presumed consent to cardiac arrest research are examined to determine if they qualify as justified infringements of the right to autonomy. The conclusion is that only presumed consent can be used, provided that the researcher can honestly say that outside of the randomized clinical trial of two or more treatments, a physician would have no basis for choosing one over the others. PMID- 8424602 TI - National health reform and emergency medicine. AB - I accept the responsibility of the office of President of ACEP today with great pride--not in myself at having achieved this prestigious office, but rather in the specialty of emergency medicine. I am honored to have the privilege and pleasure of your association and support, and I am immensely proud of being an emergency physician. I have presented these ideas and challenges in a spirit of enthusiasm and optimism for emergency medicine and ACEP. Innovation, courage, and determination are our heritage and will hopefully forever be our legacy. Emergency physicians truly are on the leading edge of the great issues of our society. I believe the time has come for us to assume a leadership position in medicine that reflects the dedication, compassion, and pioneering spirit on which this specialty was founded nearly 25 years ago--and which is so evident in this room today. Please join me in this exciting and worthwhile endeavor. PMID- 8424603 TI - Emergency medicine 2000: residencies, resident graduates, and ABEM diplomates. PMID- 8424604 TI - Moving to solve our manpower crisis. PMID- 8424605 TI - Jet travel, a pathway to emotional isolation. PMID- 8424606 TI - On-line medical command. PMID- 8424607 TI - E codes and injury surveillance in the ED: applause and warning. PMID- 8424608 TI - E codes and injury surveillance: Part 2. PMID- 8424609 TI - Out-of-hospital resuscitation research: rationale and strategies for controlled clinical trials. AB - There is no better place to test life-saving resuscitation interventions than in the prehospital setting. Patients rarely survive cardiac arrest if resuscitation techniques have failed before leaving the scene. Also, paramedics are usually very experienced in key initial resuscitative techniques, and they routinely operate under strict paramilitary protocol, resulting in better study compliance. In addition, the large study populations that are derived from emergency medical services (EMS) systems lead to faster study completion and statistically stronger data. Most important, by reinforcing standardized care, rigidly scrutinized trials improve patient care, regardless of the effect of the study intervention. The success of productive EMS research centers requires routine communication between hospital and EMS administrators and their medical directors, designation of mutually acceptable data collectors who guarantee confidentiality, reciprocal exchange of study data provided as educational seminars to the hospitals, commitments to support the budget requests of an EMS program and appropriate system modifications, inclusion of EMS personnel in study design from the very beginning, prospective education of the medical community and media before protocol implementation, an authoritative grassroots medical director, and a paramedic supervisor system. PMID- 8424610 TI - Interposed abdominal compression-CPR: a case study in cardiac arrest research. AB - When the abdomen is compressed manually in counterpoint to the rhythm of chest compression, in the performance of interposed abdominal compression-CPR, artificial circulation is approximately doubled in animal experiments and in electronic models of the circulatory system. These studies suggest that external manual compression of the abdominal aorta acts like an intra-aortic balloon pump to increase aortic pressure, whereas external manual compression of the abdominal veins acts to prime the right heart and pulmonary vessels before the next chest compression. As a result, perfusion pressures and flows are increased. Several clinical studies of this technique have shown promising results including improved hemodynamics, resuscitation success, and survival. The history of interposed abdominal compression-CPR research suggests a number of principles that may be useful in the development of other new methods for the management of cardiac arrest, including the virtues of vigorously pursuing a new idea suggested by serendipitous observations, developing and refining a working hypothesis as to pathophysiologic mechanisms, working in interdisciplinary groups, refining a novel technique in stages as experience is gained, and recognizing the need for staged phase 1, 2, and 3 clinical trials in the context of the approximately ten year gestation period from laboratory inspiration to clinical practice. PMID- 8424611 TI - Moving toward uniform reporting and terminology. AB - Researchers interested in performing research on prehospital cardiac arrest should carefully review the presentations from the session "Moving Toward Uniform Reporting and Terminology." Although each presentation concentrates on specific research topics, taken together they suggest the next evolutionary steps we should take to conduct such research. We will base these further steps on the following insights. First, all future articles on prehospital cardiac arrest must share a common nomenclature and template for reporting outcomes. The Utstein style has not solved this problem completely, but it is the critical first step. Over the coming years, we must, through continued use, progressively refine the Utstein style. Second, we can no longer depend on research that comes from a single EMS system. Although we have gained important insights from such studies in the past, our expectations of greater validity and generalizability are rising and pushing us toward multicenter, cooperative studies. The International Brain Resuscitation clinical trials and the numerous studies on thrombolytic therapy demonstrate the directions we must head. Third, we must abandon our narrow focus on the pre-hospital experience. Although some studies have avoided this problem, the preponderance of clinical studies on prehospital cardiac arrest fails to gather information on the prearrest condition of the patient, the actual decisions and action taken around the event (witness Kellermann's discussion of death criteria and rules for stopping resuscitation efforts), and the clinical experience after successful resuscitations. Fourth, we can no longer be satisfied with simple statistics on dichotomous (yes/no) survival.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424612 TI - The Utstein style for uniform reporting of data from out-of-hospital cardiac arrest. AB - The Utstein style for uniform reporting of data from out-of-hospital cardiac arrest was developed to solve a major problem in resuscitation research. Outcome measures related to cardiac arrest are difficult to evaluate or compare because there have been no uniform definitions or uniform agreements on what data to report. Widespread acceptance of the Utstein style will lead to a better understanding of out-of-hospital cardiac arrest. PMID- 8424613 TI - Challenges in cardiac arrest research. PMID- 8424614 TI - Beyond Utstein: implementation of a multisource uniform data base for prehospital cardiac arrest research. AB - This article is an overview of the issues affecting the use of a multisource uniform data base for prehospital cardiac arrest research. The goals of data base standardization include improved communication between emergency medical services (EMS) systems and between EMS physicians and the lay public; improved quantification of EMS system performance and documentation of benefit; in-depth analysis of factors that contribute to outcome; and development of EMS system models that will enhance system planning. Challenges to developing a multisource research data base include establishing a concensus approach to data base development; enlisting EMS system cooperation; verifying the accuracy of the data collected; coordinating data storage and analysis; and obtaining fiscal support for such a project. One illustrative approach to initiating multisource data base research is outlined. The iterative nature of data base development, use, expansion, and refinement is emphasized. Future reports addressing the methodology of multisource data base development and application in EMS systems should build on these concepts. PMID- 8424615 TI - Criteria for dead-on-arrivals, prehospital termination of CPR, and do-not resuscitate orders. AB - In contrast to the current consensus that governs the mechanics of prehospital advanced cardiac life support (ACLS), uniform criteria for determining when to initiate, withhold, or terminate ACLS in the field do not exist. Most emergency medical services (EMS) permit paramedics and other prehospital providers to withhold resuscitation when the victim obviously is dead, but the accuracy and appropriateness of this judgement in the field have not been subjected to empiric research. Do-not-resuscitate orders on patients in community settings often are problematic when paramedics and other prehospital providers are governed by standing orders that require them to initiate CPR when it is indicated medically. To date, eight states and a number of local EMS systems have developed a variety of policies to address this dilemma. Currently, few services permit paramedics to terminate ACLS in the field when such efforts fail to achieve return of spontaneous circulation. Studies have demonstrated convincingly that the rapid transport of such patients for further attempts at resuscitation in the hospital yields dismal rates of survival. The costs, risks, and benefits of this practice in community settings must be reviewed carefully to allocate EMS resources in an optimal manner. PMID- 8424616 TI - EMS system performance: the use of cardiac arrest timelines. AB - The time data used to analyze the significance of time-to-treatment intervals on times from sudden cardiac arrest vary by definition of time and individual system design, making it difficult to compare these data and to extrapolate findings from one study to another to achieve generalizable conclusions. A model is proposed to describe the time-to-treatment/time-to-patient response intervals sought by most researchers to evaluate quality of care and system performance. It provides a mechanism for incorporating system design elements into the cardiac arrest timeline proposed by the Utstein Conference. The present model is composed of generic patient management intervals (classes of intervals common to all systems) that are composed of component intervals (intervals particular to each system) that describe the system's conduct of an emergency medical services call, which, in turn, are constructed of data (activity) points unique to each system. Because each system defines its own data points, the present model permits rigorous definition and analysis of the time data produced by all systems. Each system thereby constructs reliable component intervals, producing a system specific, real-time depiction of the events related to a cardiac arrest call. Because all systems share the same generic patient management intervals, the resulting cardiac arrest timeline can be compared with those from other systems to assess system performance. The goal of the present model is to facilitate further the standardization of cardiac arrest data acquisition and reporting and evaluation of emergency medical services system performance. PMID- 8424617 TI - Outcomes after cardiac arrest. AB - The simplest, most easily determined, and most easily understood outcomes after cardiac arrest are survival and awakening. Awakening is defined by the patient's being able to follow commands or produce comprehensible speech. Both occur at specific times, thus lending themselves to life-table analyses. Unfortunately, these simple measures are not adequate to characterize the disability that may be present in those who survive and awaken after cardiac arrest. For such patients, measures of independence are needed. These measures often require longer follow up, direct contact with patients, and a greater understanding of the instrument to be used than for the simple measures. Investigators must decide based on the goals of a particular study what outcomes are most appropriate and the amount of resources that they are willing to devote to outcome assessment. As initial steps in resuscitation research, there may be more to gain from studies of large numbers of patients evaluated with simple measures than small numbers of patients evaluated intensively with more detailed measures. PMID- 8424618 TI - Physiologic measurement of the ventricular fibrillation ECG signal: estimating the duration of ventricular fibrillation. AB - Estimating the duration of ventricular fibrillation may help determine the best initial therapy and provide estimates of the most appropriate dose of epinephrine to administer during cardiac arrest and resuscitation. In addition, estimating this time can provide a more sound methodologic approach to stratifying patients in the analysis of cardiac arrest studies. In an initial series of studies in swine, we attempted to determine whether changes in the frequency or amplitude (power) of the ventricular fibrillation ECG signal during cardiac arrest could be used to estimate this time. These studies characterized the dynamics of both the total power and the frequency distribution of the power in the ECG over time during ventricular fibrillation in swine. Our purpose was to determine whether sufficient information existed in either parameter to estimate the duration of ventricular fibrillation. The median frequency of the power spectrum was used to track power distribution. Both parameters followed a dynamic and repeatable pattern. A mathematical model of median frequency was developed and used with data obtained from additional swine to estimate the duration of ventricular fibrillation. The model estimated the duration of ventricular fibrillation to within +/- 1.3 minutes of the actual duration from one to ten minutes after the onset of ventricular fibrillation. We recently characterized the time course of the median frequency during ventricular fibrillation in human beings. The median frequency was extracted from each four-second segment of the ventricular fibrillation ECG recordings and plotted versus time from the onset of cardiac arrest. The median frequency in human beings followed a repeatable time course during ventricular fibrillation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424619 TI - Field verification methodology using bar coding to record data. AB - Bar codes, the ubiquitous labels on consumer products, can be used to record patient care data. Bar code entry offers precise, time-stamped, data records. Bar code readers, coupled with a personal computer, can produce an electronic medical record as well as analyze data trends. PMID- 8424620 TI - Study design in cardiac arrest research: moving from the laboratory to the clinical population. PMID- 8424621 TI - Use of a cassette recorder for data collection in prehospital cardiac arrest research. AB - The methods of obtaining data and assessing protocol compliance in prehospital research can present difficulties. The Norwalk Hospital Mobile Emergency Medical Service paramedics use a minicassette recorder carried in the monitor defibrillator pack during their participation in a cardiac arrest study. The device is simple, inexpensive, and well accepted by the paramedics. With this recorder, the investigator is able to accurately identify when interventions occurred and the patients' response to therapy. The use of a minicassette recorder can facilitate data collection for prehospital research with minimal disruption for the paramedic providing care. PMID- 8424622 TI - Incidence of cardiac arrest: a neglected factor in evaluating survival rates. AB - STUDY OBJECTIVES: To add to our understanding of survival rates in out-of hospital cardiac arrest studies, we examined the incidence of cardiac arrest in the published literature. We specifically estimated if incidence rates are uniform between communities and if any relationship exists between incidence and the reported survival rates. DESIGN: A retrospective study of nearly 100 cardiac arrest peer-reviewed articles from 1970 to 1989 was performed to identify reports that included rates for incidence and survival or provided sufficient data for the calculation of these rates. MEASUREMENTS AND MAIN RESULTS: We were able to obtain reported or calculated incidence and survival rates for 20 communities. Statistical analysis was performed to compare incidence rates between communities and examine the relationship across these 20 studies between incidence rates and reported survival rates. Incidence rates ranged significantly from 35.7 to 128.3 per 100,000, with a mean of 62. Survival rates ranged significantly from 1.6% to 20.7%. Incidence rates in these communities were negatively related to survival rates; that is, as the incidence rate increased, the survival rate decreased. We determined the regression curve that describes this inverse relationship. This nomogram can be used to identify survival/incidence rate combinations that are significantly above or below average. CONCLUSION: The marked variations in incidence and inverse relationship between incidence and survival could be due to true variation in risk among the populations reported (ie, some populations may be older or sicker than others). Also, different research methodologies may create artifactual differences among studies as standards for designing studies, terminology, and reporting data have not been uniform. Therefore, these findings may reflect methodological differences and true epidemiological differences among communities. Future reports should include a method, such as an incidence/survival nomogram, to analyze survival rates while taking into account the community incidence rate of cardiac arrest. Further analysis of incidence and survival is necessary to improve intersystem comparisons, a prerequisite to sound decisions about cardiac arrest treatment, health policy, and allocation of resources. PMID- 8424623 TI - Challenges in cardiac arrest research: data collection to assess outcomes. AB - To enhance comparability in reports on survival from out-of-hospital cardiac arrest, an international task force recently developed a set of guidelines for uniform terminology, definitions, and data collection for outcome research on cardiac arrest--the Utstein style. Because the data collection recommended is limited to information available through emergency medical services systems, the potential for bias in comparisons of cardiac arrest outcomes remains. By expanding data collection to include the identification of all cases of cardiac arrest in the community, including patients who do not present for care by an emergency medical services system, a population-based approach can be achieved. We review the strengths and limitations of both emergency medical services-based and population-based data collection to assess outcomes of cardiac arrest, outline practical steps required to implement a population-based approach, and suggest that extension of the Utstein style guidelines to include all cardiac arrest cases within a defined population is needed to minimize potential bias in comparisons of cardiac arrest outcomes across communities or over time. PMID- 8424624 TI - The complexity of comparing different EMS systems--a survey of EMS systems in Europe. AB - In Europe, emergency medical care has developed since the Middle Ages in each country, even within regions of a country, resulting in a patchwork of definitions, legislations, and systems. As a consequence, emergency medical care was implemented differently according to sociocultural, geographic, political, and religious differences between and within individual European countries. The objective of this survey was to describe the emergency medical services (EMS) systems in place throughout Europe, the type and qualification of the personnel, citizen-CPR knowledge, and experiences with automated external defibrillator programs. In many European countries, the active involvement in the field of physicians, as members of the first or the second tier, was observed as a major difference between European and US EMS systems. To evaluate and to compare performance of emergency medical care in different communities, detailed knowledge of all elements of the "cardiac arrest-resuscitation complex" is required: the demographics of the community served by the EMS system, the structure and characteristics of each individual system, the epidemiology of cardiac arrest, the intervention process, and the outcome. To describe the EMS system, a uniform nomenclature is required. The Utstein "template" style could be proposed as the guideline to describe individual systems. The European Resuscitation Council could contribute in coordinating and standardizing the various aspects of emergency medical care in Europe, with detailed registration, medical coordination, and medical regulation being the principal working rules. PMID- 8424625 TI - Comparative endocrinology of molting and reproduction: insects and crustaceans. PMID- 8424626 TI - Neuroendocrine control of sex pheromone biosynthesis in Lepidoptera. PMID- 8424627 TI - Biology and control of cattle grubs. PMID- 8424628 TI - Cost, quality, accessibility--pick two out of three. PMID- 8424629 TI - Definition of perioperative nursing; patient needs. PMID- 8424630 TI - Endoscopic carpal tunnel release. A new approach to carpal tunnel syndrome. AB - Endoscopic carpal tunnel decompression is a new and effective way of relieving CTS. It shortens the recovery period and the length of time required to resume normal postoperative activity. The majority of our patients have experienced relief of symptoms and, in instances where the opposite hand was affected by CTS, have returned for that hand to be surgically repaired. Because there is less trauma with this method, and because of the patient's rapid recovery, it is common to perform bilateral releases. It is thought that patients who require surgical intervention will choose to have the procedure done sooner because of the availability of endoscopy, thereby decreasing the incidence of entrapment neuropathy that occurs late in the CTS process. PMID- 8424631 TI - Laparoscopic cholecystectomy. An electrosurgical approach to biliary disease. PMID- 8424632 TI - The bloodborne pathogens final rule. Understanding the regulation. PMID- 8424633 TI - A collaborative approach to standards, practices. Setting the stage for continuous quality improvement. AB - In retrospect, the most important thing we did was work together. We analyzed, refined, and validated our philosophical approach to patient care. We provided an information data base that is readily available for on-the-job reference and serves as a starting point for CQI activities. The very act of joint documentation of practices encourages open discussions about improvements to patient care. One physician states, We know that flaws in the process through which we produce care are everywhere--waste, duplication of effort, unnecessary complexity, and unpredictability . . . I believe that modern total quality management offers enormous hope to a medical care field that is rather desperate. . . . Collaborative practice and CQI activities are one hope. The scope of what nurses and physicians traditionally consider when discussing standards and practices must widen. We should no longer look only at patient care. We must simultaneously focus on how the management of total systems influences quality care for all patients. The CQI process, a proactive method, requires an accurate data base of information that is easily retrieved when looking for systems and individual patient care improvements. Our Computerized Collaborative Standards and Practices Manual is the reservoir for documenting practice plans developed and approved by all the disciplines involved. The process described here began with two closely knit operating room disciplines; this framework, however, offers the potential for expansion into a hospital-wide system of information organization and use. PMID- 8424634 TI - Nursing care delivery models. The perioperative environment. AB - Changes in health care on the national level are affecting perioperative nursing drastically. Perioperative nursing needs innovative approaches to guide the delivery of nursing care for patients whose operative procedures are performed outside traditional institutions. Managed care and case management are gaining in importance as more surgical procedures are performed in the outpatient arena. New minimally invasive techniques call for new models of delivering professional nursing care. Any model that is developed must ensure that professional perioperative nursing is rewarded, quality patient care provided, and surgical procedure expenses reimbursed. PMID- 8424635 TI - Attitudes toward patients with AIDS. A comparison between urban and rural perioperative nurses. PMID- 8424636 TI - Cutting costs in the OR: a case study. PMID- 8424637 TI - Physician, nursing, facility implications of informed consent. PMID- 8424638 TI - Monitoring i.v. conscious sedation, the legal scope of practice. AB - Registered nurses who are not CRNAs and are undertaking the administration and/or monitoring of patients receiving IV conscious sedation should consult their state board of nursing, undertake and document the additional education or training to monitor IV conscious sedation patients safety and document the successful achievement of competency in these services, and work with nursing, surgery, and anesthesia staff members to develop written protocols and policies. Written policies must include the provisions, if any, as prescribed by the state's board of nursing. The provisions of other state boards of nursing could be considered for possible inclusion. Policies should reflect nursing practice as described in the professional literature. Professional association position statements, such as those issued by the ANA and endorsed by AORN and other specialty associations and those issued by AANA, should be consulted. AORN's "Recommended practices for monitoring patients receiving intravenous conscious sedation" in the 1993 edition of AORN Standards and Recommended Practices for Perioperative Nursing should be incorporated into the institution policy as appropriate. PMID- 8424639 TI - States grapple with HIV/AIDS issues. PMID- 8424640 TI - Proposed recommended practices. The use and care of endoscopes. Recommended Practices Coordinating Committee. Association of Operating Room Nurses. PMID- 8424641 TI - Recommended practices. Universal precautions in the perioperative practice setting. Association of Operating Room Nurses Recommended Practices Coordinating Committee. PMID- 8424642 TI - Nondisulfide crosslinking and chemical cleavage of tubulin subunits: pH and temperature dependence. AB - Tubulin is known to be extremely unstable. The denaturation process partially involves irreversible aggregation, mediated by disulfide crosslinking. In addition, tubulin is known to undergo chemical cleavage during boiling in sodium dodecyl sulfate (SDS), a process that generates small peptides that have been mistaken for low molecular weight MAPs. Similar peptide cleavage has now been observed during two-dimensional denaturing isoelectric focusing-SDS polyacrylamide gel electrophoresis. This phenomenon has complicated interpretation of limited proteolysis studies of tubulin by subtilisin. In an effort to avoid this problem we have undertaken a detailed study of the solution conditions that promote chemical cleavage of tubulin. The cleavage reaction is found to be strongly pH, time, and temperature dependent. Nondenatured and denatured tubulin is susceptible to peptide cleavage, suggesting that primary structure is more important than secondary structure in selection of susceptible bonds. After transfer of cleavage products from an SDS gel to a polyvinylidene difluoride membrane, amino acid sequencing has confirmed cleavage at Asp-Pro bonds, at position 306 in alpha-tubulin and at position 304 in beta-tubulin. We also infer cleavage at the only additional Asp-Pro peptide bond located at position 31 in beta-tubulin. Heat-induced cleavage at Asp-Pro accounts for 5 of the 13 bands observed on SDS gels. In addition, a minor alpha-tubulin band has been sequenced from a two-dimensional gel, corresponds to cleavage at Asp-Cys located at alpha-tubulin position 200, and accounts for two additional bands observed on SDS gels. Under nondenaturing and nonpolymerizing conditions tubulin undergoes extensive intermolecular, disulfide crosslinking. At elevated temperatures and high pH, a small fraction of the crosslinking is not reduced by beta-mercaptoethanol. Disulfide-crosslinked aggregates are not suspected because carboxymethylation of tubulin does not prevent their formation. Lysinoalanine has been found by amino acid analysis and thus covalent lysine-dehydroalanine crosslinks are suspected. Dehydroalanine is formed by beta-elimination at serine and thus the presence of lysinoalanine is consistent with cleavage at Gly-Ser peptides, the most unstable serine peptide bond, and accounts for most of the remaining cleavage data. PMID- 8424643 TI - Inhibition of cytochromes P450 by nitric oxide and a nitric oxide-releasing agent. AB - The effect of nitric oxide (NO) on cytochrome P450-mediated benzyloxyresorufin and ethoxyresorufin O-dealkylase activity of rat hepatic postmitochondrial (S-9) or microsomal subfractions, or purified rat liver CYP2B1, was examined. Two distinct inhibitory phases were observed regardless of whether the NO was added prior to initiation of the reactions with NADPH or during the course of substrate turnover. The first was a reversible inhibition characterized by complete cessation of catalytic activity, the duration of which was NO concentration dependent with an IC50 in the range of 8-60 microM. This phase was followed by a second, irreversible, inhibitory phase characterized by a varying extent of recovery of activity, with inhibition ranging from < 1 to approximately 100%. The extent of this diminution in substrate conversion rate was also NO concentration dependent, with an IC50 in the range of 13-72 microM, and could be partially abrogated by the inclusion of bovine serum albumin in the reaction mixture. Lower IC50 values for both inhibitory phases were obtained in the case of benzyloxyresorufin O-dealkylase activity than in the case of ethoxyresorufin O dealkylase activity, suggesting a differential susceptibility to inhibition by NO for the two O-dealkylase activities. A nitric oxide-releasing compound ([Et2NN(O)NO]Na,DEA/NO) caused qualitatively similar inhibitory effects on benzyloxyresorufin O-dealkylase activity when added prior to the initiation of the reactions with NADPH, or during the course of substrate turnover. Based upon these results, it is possible that NO may play a role in the regulation of P450 activity in vivo. PMID- 8424644 TI - Modulation of protooncogene expression in rat aortic smooth muscle cells by benzo[a]pyrene. AB - Polycyclic aromatic hydrocarbons (PAHs), such as benzo[a]pyrene (BaP), cause vascular lesions of atherosclerotic etiology in avian and rodent species. Because the development of these lesions is associated with aberrant proliferation of vascular smooth muscle cells (SMCs), the present study was conducted to evaluate the effects of BaP on DNA synthesis and protooncogene expression in rat aortic SMCs. Subcultured cells were exposed to BaP (0.3-30 microM) for various times and processed for measurements of [3H]thymidine incorporation and c-Ha-ras or c-myc protooncogene expression. Sucrose density gradient analysis and gel mobility shift assays were employed to assess binding of BaP to intracellular proteins in the nuclear fraction and their interaction with a synthetic dioxin responsive element (DRE), respectively. BaP (0.3 and 3 microM) increased DNA synthetic rates in randomly cycling SMCs in a concentration- and time-dependent fashion. Increased DNA synthesis was also observed in synchronized SMCs treated with 3.0 microM BaP. Challenge of quiescent SMCs with 10% fetal bovine serum in the presence of BaP for 8 h enhanced serum-induced c-Ha-ras and c-myc protooncogene expression. Velocity sedimentation analysis of the nuclear fraction from SMCs treated with 0.3 microM [3H]BaP resulted in specifically bound peaks of 4.4 S and 6.5 S. The 6.5 S peak was competitively inhibited in the presence of unlabeled 10 microM 2,3,7,8-tetrachlorodibenzofuran, an aryl hydrocarbon ligand, while both peaks were eliminated in cells cotreated with 10 microM alpha-naphthoflavone (alpha-NF), an inhibitor of cytochrome P450 and an antagonist of PAH-protein interactions. alpha-NF also inhibited the induction of c-Ha-ras mRNA and DNA synthesis by BaP and the binding of BaP-Ah receptor complexes to the DRE. These results show that BaP enhances serum-induced protooncogene expression during the early part of the cell cycle in rat aortic SMCs and suggest that binding of BaP to intracellular proteins may play a role in these responses. PMID- 8424645 TI - Identification of cysteine residues involved in disulfide formation in the inactivation of glutathione transferase P-form by hydrogen peroxide. AB - We previously reported that rat glutathione transferase P-form (GST-P) is inactivated by hydrogen peroxide (H2O2). This involves formation of intra- or intersubunit disulfides, at least three extra bands with molecular masses of 21.5, 18, and 37 kDa being exhibited in addition to the native subunit band of 23.5 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions. In the present study, GST-P mutants whose cysteine residues were independently substituted with alanine (C14A, C47A, C101A, and C169A) by site-directed mutagenesis were used to identify the cysteine residues responsible for the disulfide bond formation. C14A and C169A were much more inactivated than native GST-P by 1 mM H2O2, whereas C47A and, especially, C101A appeared insensitive to H2O2. On SDS-PAGE, the 21.5-kDa band was not detected in either C47A or C101A. Hydrogen peroxide treatment of mouse GST II, highly homologous to rat GST-P but possessing glycine instead of cysteine at the 101st residue, did not result in generation of the 21.5-kDa band and was also associated with less inactivation. This band was therefore considered to be due to an intrasubunit disulfide bond between Cys-47 and Cys-101. The 37-kDa band was suggested to be due to the formation of intersubunit disulfide bonds between Cys 47 residues in different subunits. Thus the Cys-47 residue together with Cys-101 may be located in an important region for GSH binding, disulfide bond formation between these residues resulting in steric hindrance. PMID- 8424646 TI - 4-Hydroxybenzoate uptake in Klebsiella pneumoniae is driven by electrical potential. AB - The uptake of 4-hydroxybenzoic acid (4-HBA) in intact cells of a mutant of Klebsiella pneumoniae was investigated. Uptake of 4-HBA was shown to be an inducible system. This uptake system, at pH 7.0, has a high affinity for its substrate (apparent Kt = 13 microM) and a maximal velocity of 27.6 nmol min-1 mg protein-1. Competition studies with various structural analogs indicated a very narrow specificity of the 4-HBA uptake system. The transport system has been inhibited by inhibitors of energy metabolism and its activity has not been detected in the crude shock extracts. The effect of two ionophores, nigericin and valinomycin, on 4-HBA uptake with respect to the external pH has been studied. All observations indicate that 4-HBA uptake is active and energized by the membrane potential. PMID- 8424647 TI - Partial purification and characterization of a polyketide biosynthetic enzyme, 6 hydroxymellein synthase, in elicitor-treated carrot cell extracts. AB - 6-Hydroxymellein synthase, an induced polyketide biosynthetic enzyme in carrot cell extracts, was purified about 240-fold and its properties were compared with those of fatty acid synthetases. Synthetic activity of 6-hydroxymellein was inhibited in the presence of sulfhydryl reagents; however, cerulenin, a well known inhibitor of fatty acid synthetases, showed no inhibitory activity to the enzyme. Biosynthesis of 6-hydroxymellein includes an NADPH-dependent ketoreduction, and, in this reaction, the 4-pro-S-hydrogen of NADPH was specifically transferred to the compound. On the basis of stereochemical analyses of the biosynthetic process, it was concluded that the product of the ketoreduction is an optically active alcohol of R configuration. These stereo specificities of the reduction process are identical to those of beta-ketoacyl reductase in fatty acid biosynthesis which are considered to be conserved in all organisms. The synthetic rate of 6-hydroxymellein was markedly reduced when the assay was carried out with deuterium-labeled NADPH. The observed isotope effect on the catalytic rate (kH/kD) was 5.20, suggesting that this ketoreduction is one of the rate-limiting processes in 6-hydroxymellein synthesis. More than 85% of the synthetic activity was found in the soluble fraction of carrot cells, and, unlike in fatty acid synthetases in higher plants, organelle-localizing activity was not observed. PMID- 8424648 TI - Stable expression of guinea pig NADPH-cytochrome P450 reductase and monkey P4501A1 cDNAs in Chinese hamster cells: establishment of cell lines highly sensitive to aflatoxin B [corrected]. AB - To the cell line (A-15) expressing monkey P4501A1, which was developed from a Chinese hamster fibroblast cell line (CHL), a guinea pig [corrected] cDNA encoding NADPH-cytochrome P450 reductase was introduced and the transformants (designated AR-10, AR-13, and AR-18) stably expressing the reductase of increased level were established. The high-level expression of the reductase was confirmed by Northern and Western blot analyses and by measurement of the reductase activity in microsomes. The transfected clones showed 4- to 11-fold higher activity of the reductase than the parental cells. These cells showed about 9 fold higher sensitivity toward aflatoxin B1 (AFB1) cytotoxicity than A-15 cells and about 330-fold more than parental CHL cells. No difference in sensitivity was observed between CHL and the CHL-derived clones (CR-50, CR-52, and CR-68) expressing high levels of the reductase alone. Increased sensitivity of AR-10 cells was also observed with respect to sterigmatocystin cytotoxicity. Furthermore, in the assay for the induction of chromosomal aberrations, these clones were highly sensitive to AFB1. These results indicate that the level of the P450 reductase existing in parental CHL cells is not sufficient for maximal activity of monkey P4501A1. PMID- 8424649 TI - Porphobilinogen synthase from Escherichia coli is a Zn(II) metalloenzyme stimulated by Mg(II). AB - Porphobilinogen synthase (PBGS) is essential to all life forms; in mammals it is definitively established that Zn(II) is required for activity. The literature regarding the metal requirement for PBGS in other animals, plants, and bacteria neither establishes nor disproves a Zn(II) requirement. We have characterized Escherichia coli PBGS and found it to be remarkably similar to bovine PBGS. The similarities include a requirement for Zn(II), inhibition by 1,10-phenanthroline, an exceptional thermal stability, a requirement for free sulfhydryl(s) as shown by sensitivity to modification by methyl methanethiosulfonate, and the presence of tightly bound product on freshly isolated enzyme. Proton-induced X-ray emission analysis shows E. coli PBGS to contain a stoichiometric amount of Zn and no other metals. The most striking similarity between E. coli and bovine PBGS is the 13C NMR spectrum of enzyme-bound [3,5-13C]PBG; the chemical shifts of bound product are identical for both bovine and E. coli PBGS. Minor differences between E. coli PBGS and its mammalian counterpart include Km (ALA) = 1.9 mM, a pH optimum of 7.5-8, and its molar absorbtion coefficient expressed as A(0.1%)280 is 0.588. We conclude from these data that E. coli PBGS is a Zn(II)-metalloenzyme and that Zn(II) is required for catalytic activity, and propose that the mammalian and bacterial PBGS function by similar mechanisms. There is one significant difference between E. coli and mammalian PBGS. For E. coli PBGS, Mg(II) causes a twofold stimulation of the Zn(II)-induced E. coli PBGS activity; this effect is not seen for bovine PBGS. The stimulation of activity by Mg(II) mimics the effect of Mg(II) on plant PBGS, although E. coli PBGS does not contain the putative Mg(II) binding site recently revealed by Boese et al. [Q. F. Boese, A. J. Spano, T. Li, and M. P. Timko (1991) J. Biol. Chem. 266, 17060-17066]. This work lays the foundation for identification of functional amino acids based on the sequence similarities between PBGS from bacterial, plant, and mammalian sources. PMID- 8424650 TI - Resonance Raman spectroscopic evidence for an anionic flavin semiquinone in bovine liver monoamine oxidase. AB - The flavoprotein monoamine oxidase B (MAO B) from bovine liver, as isolated, has an unusual additional absorption band at 412 nm, which is similar to the absorption of its anionic flavin semiquinone form, (Fl.-), and other typical (Fl. ) flavoproteins. Denaturation of the enzyme results in the elimination of this anomalous absorption. The resonance Raman (RR) spectrum of MAO B as isolated is virtually identical to that of its dithionite-reduced (Fl.-) form. Both spectra show features similar to those of the RR spectrum of the (Fl.-) form of Aspergillus niger glucose oxidase (GO) in the region between 300 and 1700 cm-1 with 406.7 nm excitation. These features are readily distinguishable from those of oxidized flavin, neutral flavin semiquinone, and hemoprotein, strongly suggesting the presence of an (Fl.-) form in MAO B as isolated, even with preparations isolated in the absence of light. There are significant differences between the RR spectra of the (Fl.-) form of MAO B and those of GO or the published RR spectra of the (Fl.-) form of D-amino acid oxidase with excess substrate analog. At least some of these differences can be attributed to the different binding of flavin in the three enzymes. No EPR signals due to (Fl.-) are observed in MAO B as isolated. The dithionite-reduced (Fl.-) form exhibits approximately 50% less EPR signal than that expected from the absorption spectrum, which suggests a possible coupling of the (Fl.-) flavin with a paramagnetic center of unknown identity in the protein. The implications of these observations on MAO B with the current view of its catalytic mechanism are discussed. PMID- 8424651 TI - Identification of a new P450 subfamily, CYP4F1, expressed in rat hepatic tumors. AB - The expression of the rat cytochrome P450 CYP4 family was studied in hepatic tumors. In most of the primary and transplantable hepatic tumors studied, lauric acid omega-hydroxylase activity associated with the CYP4A subfamily enzymes decreased. The expression of CYP4A proteins and mRNAs in these tumors as assessed by Western and Northern blot was undetectable. However, while RNA analysis revealed the absence of 4A1, 4A2, and 4A3 mRNAs, the expression of CYP4 gene(s) was detected. A Uni-ZAP cDNA library was constructed from a 2-acetylaminofluorene induced transplantable rat hepatic tumor and screened with a CYP4 family probe. A full-length sequenced cDNA of 1977 bp isolated contained a 23-bp 5' untranslated region, a 1572-bp open reading frame, and a 382-bp 3' untranslated region. This cDNA sequence deduced amino acid sequence encodes a P450 protein having a conserved region of the CYP4 family exhibiting 44-45% amino acid identity to rat CYP4A subfamily members, 43% to human CYP4B1, 35 and 32% to insect CYP4C1 and CYP4D1, respectively. This new P450 was thus named CYP4F1. RNA blot analysis with CYP4F1 cDNA and CYP4F1-specific oligonucleotide probes revealed the expression of CYP4F1 in all tumors. This is the first example of a P450 constitutively expressed in rat hepatomas at levels exceeding those in the parental liver tissue. These results suggest that there is differential regulation of CYP4 genes during hepatic carcinogenesis. PMID- 8424652 TI - Kinetic and DNA-binding properties of recombinant human O6-methylguanine-DNA methyltransferase. AB - O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair protein that plays an important role in chemotherapy, mutagenesis, and carcinogenesis. Recombinant human MGMT was isolated from an Escherichia coli high performance expression system and purified to homogeneity. The kinetic and DNA-binding properties of the recombinant human MGMT were studied. The purified human MGMT reacted stoichiometrically with methylated DNA under second-order rate kinetics. The rate constant with normal methylated DNA was 1 x 10(9) M-1 min-1 at 37 degrees C. The binding to DNA was the rate determining step in the repair process. Approximately eight base pairs of the DNA substrate were covered by the human MGMT protein. The affinity constant for interaction of DNA to MGMT was approximately 4.7 x 10(5) M 1. The binding to methylated DNA was also examined; the binding affinity to methylated DNA was two times higher than that to unmodified DNA. The interaction with DNA induced a conformational change in the human MGMT protein as monitored by circular dichroism and fluorescence analysis. A similar conformational change was induced by both methylated and unmodified DNA. PMID- 8424653 TI - Metabolism of the insulin secretagogue methyl succinate by pancreatic islets. AB - Previous work demonstrated that methyl esters of succinate are potent insulin secretagogues in pancreatic islets, while unesterified succinate is not. This can be explained by studies reported here, which show that 14C-labeled dimethyl succinate is metabolized to 14CO2 by pancreatic islets, but that 14C-labeled succinic acid is not metabolized. Islets maintained at 1 mM glucose in tissue culture medium for 1 day lose the ability to release insulin in response to glucose and glucose metabolism is decreased 50-80%. The metabolism of dimethyl [1,4-14C]succinate and dimethyl [2,3-14C]succinate is decreased 50-60% in these incapacitated islets relative to islets maintained at 20 mM glucose. From the ratio of 14CO2 formed from dimethyl [1,4-14C]succinate, relative to that from dimethyl [2,3-14C]succinate, "acetate" ratios of 4.9-6.2 were calculated and from the ratio of 14CO2 formed from [2-14C]glucose, relative to that from [6 14C]glucose, "pyruvate ratios" of 1.6-1.7 were calculated. According to the 14CO2 ratios method, these ratios indicate that 53-66% of pyruvate derived from glucose enters the citric acid cycle via carboxylation and 34-47% enters via decarboxylation. Malic enzyme, which carboxylates pyruvate in the cytosol, was normal in islets maintained at 1 mM glucose. Previous work indicated that inhibition of glucose metabolism in islets maintained at low glucose is due to decreased net synthesis of the mitochondrial enzymes pyruvate dehydrogenase and pyruvate carboxylase [J. Biol. Chem. (1991) 266, 22392-22397], which decarboxylate and carboxylate pyruvate, respectively. Acetate (1 mM) but not pyruvate, when added to islets maintained at low glucose, increased dimethyl succinate metabolism to almost that of islets maintained at high glucose. This is consistent with a low amount of pyruvate dehydrogenase being unable to supply acetyl-CoA for condensation with oxalacetate (derived from succinate) and that the rate of the citric acid cycle could be enhanced by adding acetate which can bypass the reaction catalyzed by pyruvate dehydrogenase. PMID- 8424654 TI - Thermodynamic nonideality in macromolecular solutions: interpretation of virial coefficients. AB - Consideration is given to the interpretation of virial coefficients reflecting thermodynamic nonideality in incompressible solutions of a single macromolecular species for which there is no volume change on mixing. Expressions are presented for the concentration dependence of thermodynamic activity under conditions where either the chemical potential of solvent or the pressure is fixed, these two conditions being mutually exclusive. For the former situation, which applies to partition equilibrium procedures, the thermodynamic activity is most conveniently defined on the molar scale because the coefficients in polynomial expansions for the osmotic pressure and the activity coefficient in terms of molarity are then related to each other without the inclusion of partial molar volume terms. Under conditions of constant pressure a similar situation prevails provided that the osmotic pressure and the corresponding activity coefficient are expanded in powers of molality. In either case conversion of the virial expansions to the other concentration scale is possible, but requires the introduction of partial molar volume terms into the virial coefficients. The implications of these findings are discussed in relation to results obtained by osmometry, isopiestic measurements, equilibrium dialysis, gel chromatography, and sedimentation equilibrium. PMID- 8424655 TI - O-acetylserine(thiol)lyase from spinach (Spinacia oleracea L.) leaf: cDNA cloning, characterization, and overexpression in Escherichia coli of the chloroplast isoform. AB - The last enzymatic step for L-cysteine biosynthesis is catalyzed by O acetylserine(thiol)lyase (OASTL, EC 4.2.99.8) which synthesizes L-cysteine from O acetylserine and "sulfide." We have isolated and characterized a full-length cDNA (1432 bp) from a lambda gt11 library of spinach leaf encoding the complete precursor of the chloroplast isoform. The 1149-nucleotide open reading frame coding for O-acetylserine(thiol)lyase was in the direction opposite that of the lambda gt11 beta-galactosidase gene. The derived amino acid sequence indicates that the protein precursor consists of 383 amino acid residues including a N terminal presequence peptide of 52 residues. The amino acid sequence of mature spinach chloroplast O-acetylserine(thiol)lyase shows 40 and 57% homology with its bacterial counterparts. Sequence comparison with several pyridoxal 5'-phosphate containing proteins reveals the presence of a lysine residue assumed to be involved in cofactor binding. A synthetic cDNA was constructed, coding for the entire 331-amino-acid mature O-acetylserine(thiol)lyase and for an initiating methionine. A high level of expression of the active mature chloroplast isoform was achieved in an Escherichia coli strain carrying the T7 RNA polymerase system (F. W. Studier, A. H. Rosenberg, J. J. Dunn, and J. W. Dubendorff, 1990, in Methods in Enzymology, D. V. Goeddel, Ed., Vol. 185, pp. 60-89, Academic Press, San Diego, CA). Addition of pyridoxine to the bacterial growth medium enhanced the enzyme activity due to the recombinant protein. The extent of production is 25-fold higher than in chloroplast from spinach leaves and the recombinant protein presents the relative molecular mass and immunological properties of the natural enzyme from spinach leaf chloroplast. This work, together with our previous biochemical studies, are in accordance with a prokaryotic type enzyme for L-cysteine biosynthesis in higher plant chloroplasts. Southern blot analysis indicated that O-acetylserine(thiol)lyase is encoded by multiple genes in the spinach leaf genomic DNA. PMID- 8424656 TI - Examination of the role of tyrosine-174 in the catalytic mechanism of the Arabidopsis thaliana chitinase: comparison of variant chitinases generated by site-directed mutagenesis and expressed in insect cells using baculovirus vectors. AB - Using the catalytic mechanism of lysozyme as a paradigm for the mechanism of other enzymes that catalyze the hydrolysis of beta-1,4-glycosidic linkages, including chitinase, we have examined the effect of chemical modification with 1 ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC) on the reaction catalyzed by Zea mays chitinase. Inactivation with EDC did not result in derivatization of essential carboxylic acid residues, but resulted in the selective modification of a single essential tyrosine residue (Verburg, J. G., Smith, C. E., Lisek, C. A., and Huynh, Q. K., 1991, J. Biol. Chem. 267, 3886-3893). Here, we examine the role of the homologous tyrosine residue in the catalytic mechanism of the Arabidopsis thaliana chitinase. Tyrosine-174 of the Arabidopsis chitinase was replaced, with phenylalanine, alanine, histidine, and methionine by site-directed mutagenesis, and the variant chitinases were expressed in insect cells using baculovirus transfer vectors. A comparison of the reaction catalyzed by each of the variant enzymes indicates that substitution of another amino acid for Tyr-174 alters, but does not eliminate, enzymatic activity. Estimates of the specific activities of the variant chitinases reveal that substitution of His for Tyr-174 has a minimal effect on catalysis, the specific activities of the Phe and Met variants are approximately equivalent to each other, but are 60% the specific activity of wild type Arabidopsis chitinase, and the specific activity of the Ala variant is only 40% that of wild-type. The observation that the Arabidopsis chitinase is tolerant to mutagenesis at this position suggests that Tyr-174 does not participate directly in catalysis. PMID- 8424657 TI - Cloning and nucleotide sequence of a full-length cDNA encoding Ascaris suum malic enzyme. AB - The nucleotide sequence of a full-length cDNA encoding NAD(+)-malic enzyme from the parasitic nematode Ascaris suum was determined. The entire sequence of 2269 bases comprises a 5'-leader, a single open reading frame of 1851 bases, and the complete 3'-noncoding region of 340 bases. The first 12 amino acids of the translated sequence are hydrophobic, typical of mitochondrial translocation signals, and do not appear in the purified mature protein. The mature protein contains 605 amino acids and has a molecular mass of 68,478 Da. The amino acid sequences of tryptic peptides from the purified protein and also the N-terminal sequence show excellent correspondence with the translated nucleotide sequence. Comparison of the amino acid sequence of the ascarid protein with the human and rat liver NAD(+)-malic enzymes reveals highly conserved regions interrupted with long stretches of lesser homologous sequences. Structural motifs such as the putative nucleotide binding domains and also the malate binding site are clearly identified by alignment of the three protein sequences. PMID- 8424658 TI - Genistein inhibits insulin-stimulated glucose transport and decreases immunocytochemical labeling of GLUT4 carboxyl-terminus without affecting translocation of GLUT4 in isolated rat adipocytes: additional evidence of GLUT4 activation by insulin. AB - A recent study from this laboratory (Abler et al., J. Biol. Chem. 267, 18172 18179, 1992) showed genistein blocked insulin-stimulated glucose oxidation without affecting receptor autophosphorylation or tyrosine kinase activity. The mechanism by which genistein inhibited insulin-stimulated glucose metabolism was investigated in the present study. Insulin caused a approximately 12-fold increase in 3-O-methyl-D-glucose (3OMG) uptake compared to that of control cells. Basal and insulin-stimulated 3OMG transport was inhibited 40-60% by genistein in a concentration-dependent manner (10-100 micrograms/ml). Genistein had no effect on insulin-stimulated GLUT4 translocation from low density microsomes to plasma membranes as determined by Western blotting. These results suggested that genistein inhibited glucose transport in adipocytes by decreasing the intrinsic activity, rather than the number, of the plasma membrane-associated glucose transporters. We also previously reported that insulin treatment of adipocytes resulted in the immunocytochemically visualized unmasking of the carboxyl terminus of plasma membrane-associated GLUT4 and suggested the unmasking might be related to an insulin-induced increase in the intrinsic activity of the glucose transporter (Smith et al., Proc. Natl. Acad. Sci. USA 88, 6893-6897, 1991). In the present study, genistein decreased immunocytochemical labeling of plasma membrane-associated GLUT4 by approximately 50% in control and insulin-treated adipocytes by carboxyl-terminus antibodies but had no effect on labeling observed in an amino-terminus antibody. Since genistein did not affect the number of plasma membrane-associated GLUT4 transporters, this result supports the hypothesis that conformational changes in the glucose transporter, reflected by the ability of anti-carboxyl-terminus antibodies to bind to the transporter, may be an indication of the intrinsic activity of the plasma membrane-associated transporter. We therefore conclude that conformational changes in and activation of glucose transporters, in addition to insulin-stimulated GLUT4 translocation, play an important role in insulin-regulated glucose transport in adipocytes. PMID- 8424659 TI - Structural determination and promoter analysis of the chicken mitogen-inducible prostaglandin G/H synthase gene and genetic mapping of the murine homolog. AB - We have previously reported the isolation and characterization of a new form (PGHS-2) of prostaglandin G/H synthase (PGHS, cyclooxygenase) from chicken embryo fibroblasts. To further study the regulation and structure of the gene, we have cloned the entire chicken PGHS-2 (previously termed miPGHSch) gene with its 5' flanking region from a chicken genomic library. A genomic Southern blot showed the existence of a single PGHS-2 gene. The size of the gene was estimated at 8.9 kb through DNA sequencing and polymerase chain reaction analysis. The PGHS-2 gene was found to contain 10 exons, giving it a structure similar to that of the human PGHS-1 and murine PGHS-2 genes. The transcription start site was determined by primer extension, and the nucleotide sequence of 1.6 kb of the 5'-flanking region immediately upstream of the transcription start site was determined. The promoter sequence contained a TATA box and a variety of enhancer elements, including a serum response element, an AP-1, an NF-kappa B, and several SP-1 and AP-2 sites. Chloramphenicol acetyltransferase (CAT) assays showed that the first 158 nucleotides of the promoter efficiently drove transcription of the CAT reporter gene in serum-stimulated cells. Dexamethasone, a potent inhibitor of prostaglandin synthesis, had no effect on CAT activity, although this drug is known to markedly decrease PGHS-2 mRNA in vivo. This suggests that dexamethasone may inhibit PGHS-2 mRNA expression at the post-transcriptional level. Analysis of hamster/mouse somatic cell hybrids with radiolabeled cDNA probes demonstrated that PGHS-1 mapped to chromosome 2 and PGHS-2 mapped to chromosome 1 of the mouse genome. PMID- 8424660 TI - Oxidation of halides by peroxidases and their subsequent reductions. AB - The iodide oxidase activity and iodide-dependent pseudocatalatic activity of lignin peroxidase H2, an extracellular enzyme of the white rot fungus Phanerochaete chrysosporium, was inhibited by EDTA. The inhibition of iodide oxidase activity by EDTA was reversed at higher concentrations of iodide. Similar results were observed with a number of peroxidases. On further investigation, it was found that EDTA was decarboxylated in a reaction mixture containing a peroxidase, iodide, H2O2, and EDTA. EDTA was also decarboxylated by hypoiodite, a possible intermediate during oxidation of iodide by peroxidases. Iodide-dependent pseudocatalatic activity was increased with an increase in the concentration of H2O2 and inhibited at higher concentrations of iodide. EDTA was also oxidized by horseradish peroxidase, lactoperoxidase, and myeloperoxidase using iodide or bromide as a mediator. However, only myeloperoxidase was able to decarboxylate EDTA using chloride as a mediator. It is proposed that halide is oxidized to hypohalite by peroxidases. The hypohalite is then reduced by EDTA, H2O2, or halide. Reduction is associated with the decarboxylation of EDTA, oxidation of H2O2 to molecular oxygen, or oxidation of halide. PMID- 8424661 TI - tBOOH acts as a suicide substrate for catalase. AB - The effects of t-butyl hydroperoxide (tBOOH) on bovine liver catalase were investigated. tBOOH is accepted as a substrate of catalase and in the absence of hydrogen donors leads to a destruction of the enzyme via compound II formation. During the decomposition of this enzyme-substrate complex catalase serves as internal hydrogen donor which results in destruction of the enzyme. Evidence for this destruction is given by: a decrease of the Soret band in the uv/vis spectrum, iron release from the enzyme, decrease of the catalatic activity of the enzyme measured by oxygen release from hydrogen peroxide. Hydrogen donors like NADH and o-dianisidine have been found to protect the enzyme from destruction by tBOOH but lead to a structural alteration of the enzyme, shown by alteration of the electrophoretic mobility. In the presence of the hydrogen donor tBOOH is completely reduced to t-butanol, which is thought to proceed in a peroxidase-like reaction. PMID- 8424662 TI - Phosphorylation of calmodulin by the catalytic subunit of casein kinase II is inhibited by the regulatory subunit. AB - Casein kinase II (CKII) is composed of a catalytic subunit (alpha) and a regulatory subunit (beta) that combine to form an alpha 2 beta 2 holoenzyme. The alpha-subunit monomer is enzymatically active, albeit kinetically attenuated relative to the holoenzyme, and the addition of purified beta subunit stimulates its activity against casein (C. Cochet and E. M. Chambaz, 1983, J. Biol. Chem. 258, 1403-1406). Here we report a kinetic analysis of the phosphorylation of various protein and peptide substrates by the alpha subunit and the holoenzyme of Drosophila melanogaster CKII. We demonstrate that the alpha subunit, like the holoenzyme, is competent to phosphorylate typical physiological substrates such as the regulatory (RII) subunit of cAMP-dependent protein kinase (cAMPdPK), as well as artificial substrates such as alpha-casein and the synthetic peptide RRREEETEEE. The Km of the alpha subunit in each case is similar to that of the holoenzyme, whereas the Vmax is 5- to 60-fold lower. In contrast, calmodulin, a protein that is significantly phosphorylated by the holoenzyme only in the presence of polybasic compounds, is readily phosphorylated by the alpha subunit alone. While the Km values of the alpha subunit and the holoenzyme for calmodulin are similar, the Vmax of the alpha subunit is at least 10-fold higher than that of the holoenzyme. These results suggest that while the alpha subunit contains the necessary determinants for CKII substrate specificity, the beta subunit can either inhibit or activate it, in a substrate-dependent manner. Finally, we also demonstrate that polybasic compounds stimulate not only the holoenzyme but, to a lesser extent, the alpha subunit as well. PMID- 8424663 TI - Thyroglobulin glycosylation: location and nature of the N-linked oligosaccharide units in bovine thyroglobulin. AB - The cDNA sequence of bovine thyroglobulin (bTg) predicts 14 putative N-linked glycosylation sites. We have characterized the glycopeptides contained in a tryptic digest of bovine thyroglobulin in order to establish actual glycosylation sites. The distribution of oligosaccharide types within the known sites of N linked glycosylation has also been examined. Each glycopeptide was subjected to neutral and amino sugar analysis, as well as sialic acid analysis. Thirteen of the 14 putative N-linked glycosylation sites in bTg were confirmed as glycopeptides in the mature protein. Nine of these confirmed glycosylation sites appear to bear complex or hybrid type oligosaccharide units and four contain oligosaccharide structures in which only mannose and glucosamine were seen. Complex or hybrid type glycosylation was confirmed at asparagine residues 91, 464, 476, 834, 928, 1121, 1757, 1851, and 2232, while oligosaccharides containing only mannose and glucosamine were found at asparagine residues 1346, 1995, 2104, and 2277. Analysis of the amino acid sequence in the region of each putative glycosylation site predicts a high probability of a beta turn at 10 of the 13 sites. While glycosylation was distributed through most of the length of the thyroglobulin sequence, no oligosaccharides containing only mannose and glucosamine were found in the N-terminal half of the molecule. Several of the glycosylated peptides showed microheterogeneity and occurred in two or more discrete peaks on HPLC. A single putative site at asparagine residue 179 was not recovered in the purified glycopeptide population. PMID- 8424664 TI - Characterization of proteins in rat and human intestinal surfactant-like particles. AB - Phospholipid-rich particles isolated from the apical surface of rat enterocytes have surfactant-like properties and are enriched for intestinal alkaline phosphatase. Purified intact rat particles were used to produce antibodies in rabbits. Antiserum against the rat particle identified major proteins of 48, 68, 98, and 118 kDa on Western blots of isolated rat surfactant-like particle and did not detect any protein in rat intestinal basolateral membranes, rat brush border membranes, or human particles, but did detect a single 180-kDa protein in a preparation of rat milk fat globules, and two proteins (66 and 103 kDa) in rat pulmonary surfactant. The proteins detected on Western blot corresponded with the major proteins identified by Coomassie blue staining. Similar particles were isolated from the apical surface of human intestine and had an enzyme composition, buoyant density, and lipid content similar to those of the rat particles. Antiserum raised against the human jejunal particle detected proteins in the human particle by Western blot that were similar in size with the rat particle proteins (33, 52, 75, 82, and 118 kDa), but did not cross-react with human brush border or rat particle proteins. These studies demonstrate that the surfactant-like particles are present in human as well as rat intestinal tissue, demonstrate similar enzyme and protein content, and confirm their unique identity, distinct from apical brush border or basolateral membranes. PMID- 8424665 TI - Study of the mechanism of MF1 ATPase inhibition by fluorosulfonylbenzoyl inosine, quinacrine mustard, and efrapeptin using intermediate 18O exchange as a probe. AB - The mitochondrial F1-ATPase (MF1) is known to be largely or totally inhibited by combination or reaction with one fluorosulfonylbenzoyl inosine (FSBI), quinacrine mustard, or efrapeptin per enzyme. Measurements were made with 18O in attempt to ascertain if the weak catalytic activity remaining after exposure to excess of these reagents was due to retention of some activity by the enzyme modified by these inhibitors. Any such activity could have different characteristics that might be revealed by the distribution of [18O]Pi isotopomers formed from [gamma 18O]ATP. The MF1 inhibited by FSBI showed progressive appearance of two new catalytic pathways as inhibition proceeded. Both pathways appeared to be operative in the enzyme after one beta subunit per enzyme had been modified by FSBI. A high exchange pathway showed no detectable change as ATP concentration was lowered. The lower exchange pathway showed an increase in the amount of exchange with lowering of the ATP concentration, similar to the cooperative behavior observed with the unmodified enzyme. With excess ATP more product was formed by the low exchange pathway, showing that compulsory alternation between two catalytic sites was not retained. The behavior can be explained by the ability of the modified beta subunit to undergo binding changes similar to those occurring in catalysis, with the other two beta subunits catalyzing sluggish hydrolysis by different pathways because of the asymmetry introduced by the modification. Inhibition by quinacrine mustard also resulted in the appearance of two new pathways, somewhat similar to those from FSBI inhibition. In contrast, activity remaining with excess efrapeptin present showed only one pathway like that of the native enzyme. This can be attributed to a low equilibrium concentration of free enzyme and total inhibition of MF1 combined with efrapeptin. PMID- 8424666 TI - Purification and characterization of human biliverdin reductase. AB - Conversion of biliverdin to bilirubin is catalyzed by the cytosolic enzyme biliverdin reductase. We have purified and characterized the human liver reductase and find it to differ extensively from the previously described rat enzyme (H. Fakhrai and M. D. Maines, 1992, J. Biol. Chem. 267, 4023-4029) in its primary structure/composition, yet share kinetic properties. The human enzyme is substantially larger than the rat enzyme (approximately 41,000-42,000 versus 33,000-34,000), is dual cofactor and dual pH dependent, and requires free-SH groups. At pH 6.0-7.0 the NADH was the more effective cofactor, whereas at pH 8.5 8.75 NADPH was the preferred cofactor. The activity was inhibited by-SH reagents, 5'-dithiobis(2-nitrobenzoic acid) and p-chloromercuribenzoic acid, and protected from these reagents by cofactors and substrate. On two-dimensional electrophoresis, the purified protein resolved into four distinct isoelectric zones (pI 6.03, 5.83, 5.68, and 5.55) and two molecular weight forms (approximately 40,700 and approximately 39,600). Variants with similar pI values were detected in the purified human kidney reductase, although their relative tissue abundance varied. The tryptic map, amino acid composition, and sequence of NH2 terminus and four tryptic peptides of human reductase were compared with those of the rat. The HPLC profile and amino acid composition of the human and the rat enzymes differed vastly, and two tryptic peptides were present in the human that could not be detected in the predicted amino acid sequence of the rat enzyme. At the same time, the first 21 amino acids of the NH2 terminus of rat and human, except for the substitution of glutamic acid in human for lysine (amino acid 4) in the rat, were found identical and two peptides with 78-87% similarity to the rat reductase were found in the human reductase. Of the seven cysteine residues present in the human, four or five were titratable with 5'-dithiobis(2 nitrobenzoic acid). PMID- 8424667 TI - Investigation of the structural heterogeneity in the carbohydrate portion of a mouse monoclonal immunoglobulin A antibody. AB - A mouse immunoglobulin A monoclonal antibody was isolated from hybridoma culture fluid by affinity chromatography. Chemical analysis of the intact antibody showed a monosaccharide composition, which besides mannose also contained monosaccharides commonly found in N-linked complex type of carbohydrate structures. No N-acetylgalactosamine was found showing the absence of O-linked oligosaccharides. The carbohydrate chains were released from the polypeptide and after fractionation on immobilized concanavalin A and high-performance ion exchange chromatography structural analysis was performed. The structures were determined by chemical analyses, periodate oxidation in combination with fast atom bombardment mass spectrometry, and 500 MHz 1H NMR spectroscopy. The data revealed a great structural heterogeneity, including partially sialylated bi- and triantennary type of structures. Both types contained in addition species with branches terminated by Gal alpha 1-3Gal sequences. PMID- 8424668 TI - Oxidation of hydroquinone by copper: chemical mechanism and biological effects. AB - Exposure of humans and experimental animals to benzene has been shown to result in hematotoxicity such as pancytopenia, aplastic anemia, and leukemia. The oxidative activation of the benzene metabolite, hydroquinone (HQ), in the bone marrow to the electrophilic benzoquinone (BQ) has been suggested to play an important role in benzene-induced hematotoxicity. Since the interaction of several xenobiotics with copper has been shown to result in their metabolism, in this study we have investigated the role of copper in the oxidation of HQ and HQ induced toxicity to mice bone marrow stromal cells, target cells of HQ in the bone marrow. In phosphate-buffered saline, HQ underwent autoxidation slowly to BQ, while the presence of Cu(II) ions (1, 2.5, 5, 10, 50 microM) strongly accelerated the oxidation of HQ to BQ in a concentration-dependent manner. Reaction of HQ with Cu(II) was also accompanied by the reduction of Cu(II) to Cu(I), the utilization of O2, and the concomitant generation of H2O2. The oxidation of HQ by Cu(II) could be blocked by the Cu(I)-specific chelator bathocuproinedisulfonic acid (BCS), particularly when the ratio of BCS to Cu(II) was 4:1. By observing the kinetics of the reactions derived from mixing 100 microM HQ and 100 microM Cu(II), it was found that all of the Cu(II) was reduced to Cu(I) within 5 s, followed by consumption of O2 and the generation of BQ, which reached maximum levels at 4 min after mixing HQ and Cu(II). In addition, oxidation of HQ by Cu(II) also generated chemiluminescence. In the presence of myeloperoxidase, Cu(II)-mediated oxidation of HQ was increased. Addition of Cu(II) to primary bone marrow stromal cell cultures significantly enhanced HQ induced cytotoxicity. The enhanced cytotoxicity of HQ by Cu(II) could be completely prevented by adding BCS, glutathione (GSH), or dithiothreitol but not by catalase. Supplementation of stromal cells with 20 microM BCS in the absence of exogenously added Cu(II) significantly abated HQ-induced cellular GSH depletion and cytotoxicity, suggesting a possible involvement of endogenous copper in the activation of HQ. The above results indicate that Cu(II) strongly induces the oxidation of HQ and as such may be a factor involved in the oxidative activation and toxicity of HQ in target cells. PMID- 8424669 TI - Adriamycin conjugates of human transferrin bind transferrin receptors and kill K562 and HL60 cells. AB - Adriamycin (ADR) was coupled to human transferrin (TRF) by using a glutaraldehyde crosslinking method. The TRF-ADR conjugates were separated by column chromatography and the molar ratio of ADR to TRF (i.e., conjugation number) for the studied conjugates was found to be 1.2. Analysis in sodium dodecyl sulfate polyacrylamide gels demonstrated that TRF-ADR conjugates with this molar ratio had the same mobility as native TRF and contained few aggregates. The ADR remained conjugated to TRF under conditions of decreased pH known to occur in many intracellular compartments, and analysis by spectrofluorometry revealed that the conjugated ADR retained its ability to intercalate DNA. The TRF-ADR conjugates were shown by flow cytometry to preferentially bind tumor cells and cell-bound conjugates were found to be laterally mobile within plasma membranes. The binding of TRF-ADR conjugates was determined to be saturable, and competition experiments done with both radioiodinated and fluorescein-labeled TRF-ADR conjugates demonstrated dose-dependent inhibition of conjugate binding by unlabeled TRF, indicating that TRF-ADR conjugates were bound by TRF receptors. Cytotoxicity studies performed with tritiated thymidine incorporation and tetrazolium reduction assays revealed that TRF-ADR conjugates inhibited the proliferation of both K562 and HL60 cells in culture more effectively than free ADR. Such conjugates could provide a delivery system for ADR that would target the drug and possibly diminish its dose-associated complications. PMID- 8424670 TI - Role of flavin in acetoin production by two bacterial pyruvate oxidases. AB - Escherichia coli pyruvate oxidase (POXEC) requires FAD both for the oxidative decarboxylation of pyruvate to acetate and CO2 and for the formation of acetoin from pyruvate and acetaldehyde. Prior work has shown that the catalytic activity (kcat/Km) for POXEC in the oxidative reaction is stimulated approximately 450 fold by amphiphilic activators. This paper shows that the acetoin reaction does not respond to activation. The FAD requirement for acetoin formation can be replaced by 5-deaza-FAD and 6-hydroxy-FAD, FAD analogs which form kinetically stable oxidized and reduced enzyme species, respectively. As would be expected, the 5-deaza- and 6-hydroxy-FAD enzymes are not active in the oxidative reaction. A second flavin pyruvate oxidase from Pediococcus pseudomonas (POXPP), which catalyzes the oxidative decarboxylation of pyruvate to CO2 and acetyl phosphate, also requires FAD for acetoin formation. POXPP has an oxidative rate comparable to that of POXEC, but in comparison to POXEC, POXPP catalyzes acetoin formation at a much reduced rate. Again, as was found with the POXEC, an FAD analog incapable of undergoing facile oxidation-reduction reactions also could replace the FAD requirement in the POXPP acetoin reaction. The results indicate that the role for FAD in acetoin formation with both enzymes is based on a structural requirement and that FAD does not participate in a redox function in the acetoin reaction. PMID- 8424671 TI - Role of membrane defects in the regulation of the activity of protein kinase C. AB - With mixtures of phosphatidylethanolamine and phosphatidylserine, the activity of protein kinase C is highly dependent on the fatty acid composition of the phosphatidylethanolamine. This contrasts with phosphatidylcholine/phosphatidylserine mixtures which affect the activity of PKC in a manner independent of the fatty acid composition of the phosphatidylcholine. The results are in accord with those phospholipids having the lowest bilayer hexagonal phase transition temperature being most effective in augmenting the activity of PKC in the presence of its cofactors. Although the activity of this enzyme is markedly sensitive to the presence of hexagonal phase forming lipids, the activity is insensitive to differences between gel and liquid crystalline state membranes. Membrane defects alone also do not explain the observed effects since vesicles with phase boundary defects do not activate PKC. Increased hexagonal phase propensity of the lipid does not alter the partitioning of PKC between aqueous and membrane phases, which remains calcium dependent. The results demonstrate that simply the formation of defects is not sufficient to promote PKC activity, but that changes in membrane bilayer properties related to hexagonal phase propensity are required. PMID- 8424672 TI - Regulation of prostatic acid phosphatase expression and secretion by androgen in LNCaP human prostate carcinoma cells. AB - The expression and the secretion of human prostatic acid phosphatase (PAcP), a differentiation antigen which is the major acid phosphatase in prostate epithelial cells, are thought to be regulated by an androgen. We investigated this regulatory mechanism at the post-transcriptional level in LNCaP human prostate carcinoma cells using a cDNA clone for the secretory form of PAcP. 5 alpha-Dihydrotestosterone (DHT, an active form of endogenous androgen) stimulated the secretion of PAcP from cells grown in a steroid-reduced medium and in a defined serum-free medium, respectively. The secreted PAcP activity was increased following a DHT dose in a dose-dependent fashion at concentrations of up to 1 microM. Further, the stimulation of PAcP secretion occurred following the period of exposure to DHT. During a 5-day treatment period, with 10 nM of DHT in the steroid-reduced medium, the secretion of PAcP was stimulated approximately 150% over that from control cells. Nevertheless, PAcP was secreted from cells grown in the absence of added DHT. First, the androgen dependency of PAcP expression was examined. The expression and the secretion of PAcP were observed in cells that were grown in a defined serum-free medium and grown in a steroid-reduced medium, in the absence of DHT. The increased secretion by DHT was further demonstrated to be in part due to an increase in PAcP mRNA level, as evidenced by Northern blot analysis. PAcP mRNA levels were elevated approximately 2-fold and corresponded to an increase of approximately 2.5-fold in the secreted level of newly synthesized 35S-PAcP. Then, the effect of DHT on the secretory process was investigated. Results of pulse-chase labeling experiments indicated that the secretory rate of PAcP was stimulated by about 50% on average by DHT. In conclusion, our data demonstrated that, in LNCaP cells, the expression and the secretion of PAcP regulated by androgen are apparently hormone-responsive processes. Further, DHT stimulation of PAcP secretion operates within at least two levels: increased PAcP mRNA and stimulation of the secretory pathway. PMID- 8424673 TI - Overproduction of soluble trichodiene synthase from Fusarium sporotrichioides in Escherichia coli. AB - Trichodiene synthase is a sesquiterpene cyclase isolated from various fungal species which catalyzes the cyclization of farnesyl diphosphate (FPP) to trichodiene. The trichodiene synthase gene (Tox5) of Fusarium sporotrichioides has previously been cloned and expressed as 0.05-0.1% of total cell protein in Escherichia coli. We have used polymerase chain reaction to amplify the trichodiene coding sequence carried on the plasmid pTS56-1. The resulting DNA, carrying a BamHI restriction site and the T7 gene 10 ribosome binding site and translational spacer element immediately upstream of the ATG start codon as well as a HindIII site adjacent to the translational stop codon, was inserted into the corresponding sites of the expression vector pLM1. The latter vector carried the promoter and translational leader sequence from T7 gene 10 and the E. coli rmBT1T2 tandem transcription terminator. This construct was cloned into E. coli BL21 (DE3). The resulting transformants, when induced with isopropyl beta-D thiogalactoside, produced trichodiene synthase as 20-30% of total soluble protein. The recombinant synthase, which could be purified five-fold to homogeneity by ammonium sulfate precipitation, ion-exchange chromatography on Q Sepharose, and gel filtration on Superose 12, was identical to native protein in steady-state kinetic parameters and mobility on sodium dodecyl sulfate polyacrylamide gel electrophoresis and had the expected MENFP N-terminal sequence. PMID- 8424674 TI - Experimental strategy to study the pH dependence of the kinetic behavior of enzymes: practical application to chicken liver xanthine dehydrogenase. AB - An experimental strategy has been developed to analyze the pH dependence of kinetic parameters in two-substrate enzyme kinetics based upon model discrimination, parameter refinement, and optimal discrete design. The experimental protocol proposed includes obtaining reproducible experimental results in different sessions, replicates of each experimental determination, and rejection of outliers. This strategy leads to an empirical kinetic equation that fits the experimental results. As a practical application of this strategy we studied the pH dependence of kinetic parameters of chicken liver xanthine dehydrogenase. We obtained an empirical rate law from the determination of the initial rates at different substrate concentrations and pH. Finally, we have proposed a plausible kinetic mechanism that may explain the experimental findings. This model is not the only one leading to the empirical equation, but it is based on previous experimental findings (Bruguera, P., Lopez-Cabrera, A., and Canela, E. I., 1988, Biochem. J. 249, 171-178). PMID- 8424675 TI - Thiol-dependent metal-catalyzed oxidation of bovine lens aldose reductase. I. Studies on the modification process. AB - Bovine lens aldose reductase (alditol: NADP+ oxido-reductase, EC 1.1.1.21) undergoes a thiol-dependent oxidative modification catalyzed by the Fe(II)/Fe(III) redox system. The enzyme is inactivated by various oxygen radical generating systems. However, addition of 2-mercaptoethanol to the oxygen radical generating systems resulted in an initial increase followed by a decrease in the activity of aldose reductase. The net maximal increase in the enzyme activity was observed with 3 mM 2-mercaptoethanol, 0.3 mM FeSO4, and 0.9 mM EDTA, either with or without 1 mM hypoxanthine and 37 mU/ml of xanthine oxidase. The formation of the stable, activated intermediate, ARa, appears to proceed through the reaction between the enzyme and the oxidized form of 2-mercaptoethanol which in the presence of iron, forms a mixed disulfide with a cysteine residue. Reduction of ARa with dithiothreitol released 0.7 mol of 2-mercaptoethanol per mole of enzyme and converted it to a form that resembled the native aldose reductase. PMID- 8424676 TI - Thiol-dependent metal-catalyzed oxidation of bovine lens aldose reductase. II. Proteolytic susceptibility of the modified enzyme form. AB - Bovine lens aldose reductase (alditol: NADP+ oxidoreductase, EC 1.1.1.21) undergoes a modification induced by 2-mercaptoethanol in the presence of the redox system Fe(II)/Fe(III). The modified form (ARa) exhibits an increased hydrophobicity and tendency to aggregate. Moreover, while the native enzyme form is rather insensitive to proteolytic breakdown, the modified form is susceptible to limited proteolysis by trypsin and chymotrypsin. With both proteases, the degradation correlated with a loss of enzyme activity and results in the appearance of one molecular species of 26 KDa (for chymotrypsin) and two molecular species of 24 and 17 KDa (for trypsin). The decline in solubility and the increase in susceptibility to proteolysis of ARa suggests that the thiol dependent metal-catalyzed modification is comparable to other oxidative systems that mark proteins for degradation. PMID- 8424677 TI - Bis-allylic hydroxylation of polyunsaturated fatty acids by hepatic monooxygenases and its relation to the enzymatic and nonenzymatic formation of conjugated hydroxy fatty acids. AB - [14C]Linoleic acid was incubated with phenobarbital-induced rat liver microsomes and formation of cis-trans-conjugated hydroxy fatty acids was investigated. 13 Hydroxy-9Z,11E-octadecadienoic acid (13-HODE), 9-hydroxy-10E,12Z-octadecadienoic acid (9-HODE), and three novel metabolites were identified, viz. 11-hydroxy 9Z,12Z-octadecadienoic acid (11-HODE), 8-HODE, and 14-HODE. 11-HODE (59% R), the main product, was unstable and converted to 9(R, S)-HODE and 13(R, S)-HODE in acidic media. All metabolites contained oxygen from O2. Experiments under oxygen 18 gas showed that 13-HODE and 9-HODE contained equal or less amounts of oxygen 18 than the other metabolites. In the former case, 9-HODE and 13-HODE were formed with stereo-selectivity (80-82% R). [11S-2H]Linoleic acid was metabolized to 13R HODE with loss of deuterium (24% 2H) and to 9R-HODE with deuterium retention (95% 2H), while [11R-2H]linoleic acid was metabolized to 13R-HODE that largely retained the label (71% 2H) and to 9R-HODE that lost most of the label (22% 2H). These data indicated that P450 catalyzed abstraction of the pro-R hydrogen at C11, double bond migration and suprafacial oxygen insertion at C9 to give 9R HODE, while abstraction of the pro-S hydrogen at C11, followed by double bond migration and oxygen insertion, yielded 13R-HODE. Hepatic microsomes of the cynomolgus monkey metabolized 18:2n-6 as above and 20:4n-6 to 13 hydroxyeicosatetraenoic acid, likely formed in analogy with 11-HODE. In summary, one mechanism in the biosynthesis of cis-trans-conjugated hydroxy fatty acids by P450 involves suprafacial hydrogen abstraction and oxygen insertion. In addition, hydrolysis of the unstable bis-allylic hydroxy metabolites may contribute to the formation of conjugated hydroxy fatty acids. PMID- 8424678 TI - Identification of deoxyribonuclease II as an endonuclease involved in apoptosis. AB - Cell death occurs by apoptosis during programmed deletion of cells and following exposure to cytotoxic agents. Central to the mechanism of apoptosis is internucleosomal DNA digestion by an endogenous endonuclease which is thought to mediate cell death. An axiom of apoptosis is that the endonuclease involved is a Ca2+/Mg(2+)-dependent endonuclease. During purification of endonucleases from Chinese hamster ovary cells, we found little Ca2+/Mg(2+)-dependent endonuclease activity, but large amounts of an endonuclease active below pH 7. This acidic endonuclease was activated in intact cells by reducing intracellular pH values below 7 with a proton ionophore. This activity generated internucleosomal digestion of DNA characteristic of apoptosis. Nuclear extracts contained a cation independent endonuclease with identical pH-dependent activity. We have compared the acidic endonuclease to bovine deoxyribonuclease II (DNase II) and have found them nearly identical by all tests, including sensitivity to various inhibitors, purification by the same chromatographic steps, and recognition by antibody raised against the bovine enzyme. Addition of either the acidic endonuclease or bovine DNase II to isolated nuclei induced internucleosomal DNA digestion up through pH 6.5. These data demonstrate that DNase II can mediate internucleosomal DNA digestion characteristic of apoptosis following intracellular acidification. Furthermore, these data question the premise that the Ca2+/Mg(2+)-dependent endonuclease is the only endonuclease involved in apoptosis. PMID- 8424679 TI - Structural and functional studies on the interaction of sodium dodecyl sulfate with beta-galactosidase. AB - The effect of sodium dodecyl sulfate (SDS) on enzyme activity, electrophoretic behavior, and conformation of Escherichia coli beta-galactosidase is presented. Fourier-transform infrared spectroscopy (FT-IR), previously used to study the structure of native beta-galactosidase has been applied to examine the detergent effects on the enzyme. At 20 degrees C, the presence of 1% SDS does not cause appreciable changes in the secondary structure, and enzyme activity is preserved; however, 10% SDS produces complete enzyme inactivation and FT-IR spectroscopy indicates a concomitant change in conformation. Thermal denaturation of beta galactosidase starts at approximately 53 degrees C in the absence and at approximately 46 degrees C in the presence of 1% SDS, indicating tertiary structure changes; also, a good correlation between structural (FT-IR) and functional (Arrhenius plots) data is observed. The secondary structure of thermally denatured beta-galactosidase contains mainly extended structures, and intermolecular interactions produce protein aggregation. In the presence of 10% SDS, however, the hydrophobic segments of the protein are stabilized by SDS into helical structures without protein aggregation. At 30 degrees C, in the presence of 1% SDS, two protein bands are resolved by gel electrophoresis, only one of them being active. A model for SDS-galactosidase interaction is proposed, according to which, at low surfactant concentrations, SDS molecules bind the outer surface of the protein, without affecting the protein core. Higher detergent concentrations produce a larger conformational change involving enzyme inactivation and increased accessibility of the solvent to the protein core. Increasing temperature in the presence of 10% SDS leads to a facilitated access of surfactant molecules to the inner protein regions and to an increase of the beta-galactosidase alpha-helical content. PMID- 8424680 TI - Effects of cyclic hydrostatic pressure on proteoglycan synthesis in cultured chondrocytes and articular cartilage explants. AB - Primary chondrocyte cell cultures and explants of bovine articular cartilage were subjected to cyclic hydrostatic pressure in a novel computer-controlled pressure chamber designed for this purpose. The cultures were labeled with 5 microCi/ml 35SO4 and simultaneously pressurized with 5 MPa load for 1.5 or 20 h with pressure cycles of 0.0167, 0.05, 0.25, and 0.5 Hz. The chondrocyte cell cultures were also subjected to 0.0082 and 0.0034 Hz cycles. Sulfate incorporation was significantly inhibited in cell cultures subjected to the 0.5, 0.25, or 0.05 Hz cyclic loads for 1.5 h, but stimulated in explant cultures with a 0.5 Hz cyclic 1.5-h load. Chondrocyte cultures subjected to longer (20 h) loading showed a stimulation of sulfate incorporation with 0.5 and 0.25 Hz cycles, but an inhibition with 0.0167 Hz. The results indicate that cyclic hydrostatic pressures of presumably physiological magnitude have significant influences on proteoglycan synthesis in articular cartilage chondrocytes. Comparison of the cell and explant cultures under identical pressure conditions suggested that chondrocyte interactions with extracellular matrix are involved in this regulation by cyclic hydrostatic pressure. The responses of the chondrocytes to pressurization also varied according to the total length of the treatment, a finding compatible with the idea of multiple metabolic steps in chondrocytes, both pre- and post translational, controlled by the ambient hydrostatic pressure. PMID- 8424681 TI - Isolation of two pyruvate kinase activities in the parasitic protozoan Leishmania mexicana amazonensis. AB - Using phosphocellulose affinity chromatography we were able to separate two pyruvate kinase (EC 2.7.1.40) activities in the parasitic protozoan Leishmania mexicana amazonesis. One activity (PYK1) showed hyperbolic kinetics and was decreased by fructose 2,6-bisphosphate, whereas the second activity (PYK2) showed sigmoidal kinetics for the substrate phosphoenolpyruvate and was activated by fructose 2,6-bisphosphate. Molecular sieve chromatography (Sephacryl S-400) of PYK1 produced a single peak of apparent molecular mass around 200,000, while PYK2 eluted at a position corresponding to M(r) 55,000. PMID- 8424682 TI - The effector roles of kringle 1 and kringle 2 in the enzymatic properties of recombinant tissue-type plasminogen activator as revealed by generation of recombinant molecules containing each kringle linked to the protease domain. AB - Recombinant DNA technology has been employed to construct and express in human kidney 293 cells cDNAs encoding deletion-mutant recombinant (r) tissue-type plasminogen activators (tPA) retaining only the kringle 1 ([K1tPA]) and serine protease (P) functional domains (r-[K1tPA]P), and the kringle 2 ([K2tPA]) and P domains (r-[K2tPA]P), along with a variant of r-tPA containing a W253S mutation (r-tPA/W253S). Of these mutants, only r-[K2tPA]P retained its ability to interact with omega-amino acid effector molecules. The Km for single-chain wild-type (wt) r-tPA toward the synthetic substrate H-D-Ile-L-Pro-L-Arg-p-nitroanilide (S2288) was decreased by approximately 3-fold in the presence of a saturating concentration of human fibrinogen (Fg), along with a small (1.14-fold) increase in the kcat for this same reaction. The kinetic activation (dissociation) constant (KA) for Fg was 2.4 microM. Fg did not influence the steady-state amidolytic properties of two-chain wtr-tPA. Similar effects of Fg on the Km for hydrolysis of S2288 were displayed for single-chain forms of r-[K1tPA]P, r [K2tPA]P, and r-tPA/W253S, with additional small effects of Fg on the kcat of this reaction. The KA for Fg toward these proteins ranged from 2.4 microM for wtr tPA to 5.2 microM for r-[K1tPA]P. The amidolytic properties of the two-chain forms of these variants were also unaffected by Fg. The activation rate of [Glu1] plasminogen ([Glu1]Pg) by wtr-tPA was stimulated approximately 7-fold by Fg and approximately 139-fold by the same concentration (in Fg equivalents) of human fibrin (Fn) (Fn/Fg stimulatory ratio = 19.9). The Fn/Fg ratio was 10.6, 20.9, and 18.0 for r-[K1tPA]P, r-[K2tPA]P, and r-tPA/W253S, respectively. Quantitative [Glu1]Pg-enriched clot lysis assays revealed that r-[K1tPA]P, r-[K2tPA]P, and r tPA/W253S were approximately 18, 72, and 54%, respectively, as effective as wtr tPA in catalyzing the plasminogen activation event leading to lysis. The antifibrinolytic agent epsilon-aminocaproic acid, inhibited clot lysis with approximately equal effectiveness in [Glu1]Pg-enriched clots when wtr-tPA, r [K1tPA]P, r-[K2tPA]P, or r-tPA/W253S were employed as the activators. These studies demonstrated that Fg- and Fn-based stimulatory effects on the enzymatic properties of r-tPA and its variants were generally not proportional to the macroscopic binding abilities of these proteins with omega-amino acids or with Fg and Fn.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8424683 TI - Purification and characterization of Clostridium difficile glutamate dehydrogenase. AB - Recombinant Clostridium difficile glutamate dehydrogenase (L-glutamate:NAD oxidoreductase, EC 1.4.1.2) was purified 177-fold to electrophoretic homogeneity with a 62% recovery through a four-step procedure involving gel filtration and ion-exchange and dye affinity chromatography. The approximate molecular weights of the native enzyme by gel filtration and subunits by sodium dodecyl sulfate polyacrylamide gel electrophoresis were consistent with a hexameric structure for the purified enzyme. The enzyme-catalyzed glutamate oxidation was an NAD dependent sequential process in which NADP could not be substituted as coenzyme. Several dinucleotide analogs of NAD structurally altered in either the pyridine or the purine moiety were observed to function as coenzymes when substituted for NAD. Nicotinamide mononucleotide did not serve as a coenzyme for glutamate oxidation. Product inhibition by NADH was competitive with respect to NAD. In deadend inhibition studies, adenosine diphosphoribose was shown to be an effective coenzyme-competitive inhibitor. PMID- 8424684 TI - The regulatory properties of kidney pyruvate dehydrogenase complex components. AB - The activities of the enzyme components of the pyruvate dehydrogenase complex are affected to different extents by changes in ionic strength and pH. At pH 7.4 the optimum activity of pyruvate dehydrogenase (E1), dihydrolipoamide acetyltransferase (E2), and dihydrolipoamide dehydrogenase (E3) occur in the ranges of ionic strengths of 0.06-0.08, 0.01-0.02, and 0.10-0.15 M, respectively. The activity of dihydrolipoamide dehydrogenase is least sensitive to changes in the ionic strength of the assay medium. At constant ionic strength (0.15 M) the optimum activity of E1, E2, and E3 occur at pH 7.4, 7.0, and 8.0, respectively. Changes in pH mostly affect the dihydrolipoamide acetyltransferase activity. Cl- and HCO3- anions inhibit the activity of pyruvate dehydrogenase. In the presence of 80 mM Cl- or HCO3- ions the activity of E1 is inhibited by 25 and 10% respectively. K+, Na+, and HPO4(2-) ions affect the activity of dihydrolipoamide acetyltransferase. The activity of this enzyme component is stimulated by 28 and 25% in the presence of 80 mM K+ and Na+ cations, respectively. HPO4(2-) stimulates the dihydrolipoamide acetyltransferase in a calcium-dependent manner. In the presence of 20 mM HPO4(2-) the activity of the dihydrolipoamide acetyltransferase increases 20 and 40% in the absence and presence of 0.1 mM Ca2+, respectively. The activity of dihydrolipoamide dehydrogenase is not affected by K+, Na+, HPO4(2-), Cl-, or HCO3-. PMID- 8424685 TI - Isozymes of lignin peroxidase and manganese(II) peroxidase from the white-rot basidiomycete Trametes versicolor. I. Isolation of enzyme forms and characterization of physical and catalytic properties. AB - The basidiomycete Trametes versicolor is a white-rot fungus and a potent degrader of lignin. The development of extracellular enzyme activities in the fungal culture under physiological conditions of secondary metabolism was investigated. Using the culture medium as starting material a large number of peroxidase forms were purified by the use of chromatographic techniques. Sixteen forms of lignin peroxidase and five forms of manganese(II) peroxidase were separated and the majority of these enzymes was characterized with respect to isoelectric point, molecular mass, and specific enzyme activity. The manganese(II) peroxidases showed a lower isoelectric point (pI 3.2-2.9) and a slightly higher molecular mass (44-45 kDa) than the lignin peroxidases (pI 3.7-3.1, and 41-43 kDa). Specific enzyme activities for the forms of lignin peroxidase, using veratryl alcohol as the substrate, were found to differ considerably. Certain differences in the specific enzyme activity were also observed among the forms of manganese(II) peroxidase. A multitude of peroxidase forms has previously been encountered in another white-rot fungus, Phanerochaete chrysosporium. The discovery that it also occurs in T. versicolor would suggest that this multiplicity could be a common feature among white-rot fungi and may be essential for the biodegradation of lignin. PMID- 8424686 TI - Modulation of glutathione S-transferase activity by a thiol/disulfide exchange reaction and involvement of thioltransferase. AB - Low concentrations of cystamine and cystine inactivated human placenta glutathione S-transferase (GST-pi) in cytosolic fraction very effectively, as did the purified enzyme, through the thiol/disulfide exchange reaction. Mixed disulfide formation of GST-pi in cytosol was prevented by thioltransferase existing in cytosol with a low concentration of GSH. This protection of GST-pi activity was more effective with the participation of glutathione reductase. The incorporation of half-[14C]cystine into a GST-pi molecule according to the inactivation was provided by autoradiography. Purified human placenta thioltransferase (1900-fold from cytosol) could release the incorporated half [14C]cystine from a GST-pi molecule with restoration of enzyme activity. Thioredoxin in placenta cytosol could not protect the GST-pi activity from inactivation at all. PMID- 8424687 TI - Studies on fibronectin and its domains. I. Novel recombinant cell-binding domain of fibronectin--a modulator of human platelet functions. AB - The DNA sequences encoding for two proteins of the cell-binding domain (CBD) of human fibronectin (FN), namely a 33-kDa protein (aa 1329-1722) and a 40-kDa (aa 1380-1851) protein, were cloned and expressed in Escherichia coli. The interactions of the resulting rCBD proteins, refolded and purified to homogeneity, with human platelets were studied in comparison with those of the pentapeptide GRGDS. The binding of both the 33-kDa and the 40-kDa proteins to washed platelets appeared to be dependent upon platelet activation. In the case of the 33-kDa protein, binding to stimulated platelets was shown to be saturable, with Kd = 2 microM (thrombin as agonist). Moreover, both the 33-kDa and the 40 kDa proteins inhibited fibrinogen binding (at 0.1 microM) to ADP- or thrombin stimulated platelets with IC50 values in the same concentration range. Binding seemed therefore to occur mainly at the GPIIb/IIIa receptor, and accordingly monoclonal antibodies against this receptor prevented up to 85% of the binding of the 33-kDa protein to platelets. With most stimuli the 33-kDa and the 40-kDa proteins inhibited platelet aggregation at concentrations 15- to 25-fold lower than those required by GRGDS and, in the case of the 33-kDa protein, this was shown to occur in either platelet-rich plasma, washed platelets, or whole blood. The 33-kDa protein also inhibited platelet aggregation and thromboxane A2 (TXA2) generation on the subendothelial extracellular matrix, whereas the GRGDS peptide inhibited only matrix-induced platelet aggregation, but not TXA2 formation. Furthermore, the 33-kDa protein, which is derived from the human FN CBD, seemed to be highly selective, since it inhibited the aggregation of platelets from primates only, and not from other animals tested. Finally, the 33-kDa protein did not promote fibroblast cell attachment, as was observed for both whole FN and the 40-kDa protein, thus displaying a selectivity toward platelets. In conclusion, the unique properties of the 33-kDa protein, and, in particular, its special affinity directed only toward activated primate platelets, seem to hold a promising potential for the further development of an antithrombotic agent. PMID- 8424688 TI - Aluminum has both oxidant and antioxidant effects in mouse brain membranes. AB - The in vitro effects of aluminum (Al) on lipid peroxidation were studied in mouse brain homogenates and purified brain subcellular fractions. In brain homogenates prepared in 5 mM Na2HPO4, 0.14 M NaCl, pH 7.4, the addition of Al decreased Fe(2+)-induced lipid peroxidation, measured as 2-thiobarbituric acid-reactive substances (TBARS), in a dose dependent manner, with a maximum effect at 250 microM Al. In brain homogenates prepared in 20 mM Tris-HCl, 0.14 M NaCl, pH 7.4, Al acted as a prooxidant at 250 and 500 microM concentrations. The prooxidant effect of Al was enhanced with increasing concentrations of Fe2+. In brain microsomes Al increased TBARS production and conjugated dienes formation, both depending on the addition of Fe2+. In myelin, the prooxidant effect of Al on Fe(2+)-induced lipid peroxidation was eliminated when membranes were disrupted with 0.2% Triton X-100. Thus, in brain homogenates, microsomes, and myelin, Al has the potential for exhibiting both prooxidant and antioxidant activity depending on the concentration of Fe2+ and Al in the media and on membrane integrity. Similar to Al, Be2+ and La3+ had prooxidant effects on Fe(2+)-induced lipid peroxidation in myelin, suggesting that membrane damage secondary to induced lipid peroxidation may be a common mechanism underlying tissue pathology even with metals without redox capacity. Oxidative damage to brain cell components may be an important mechanism mediating the neurotoxicity of Al. PMID- 8424689 TI - Glucose phosphorylation in Helicobacter pylori. AB - Saccharide kinase activities in Helicobacter pylori were investigated by incubating bacterial lysates with ATP and mono- or disaccharides and monitoring directly the appearance of phosphorylated products using 13C or 31P nuclear magnetic resonance spectroscopy. The monosaccharides employed included two trioses, two tetroaldoses, one tetroketose, five aldopentoses, two ketopentoses, five aldohexoses, three ketohexoses, and gluconic and glucuronic acids; the disaccharides studied were maltose, trehalose, cellobiose, sucrose, lactose, gentobiose, and melibiose. D-Glucose was the only sugar phosphorylated among all the carbohydrates examined. The kinase activity of lysates was studied by measuring the rates of formation of glucose 6-phosphate. The substrate specificity, the relatively high KM, and the absence of substrate inhibition suggested that the enzyme is a glucokinase rather than a hexokinase. Most of the glucose kinase activity was observed with the pellet fraction obtained by centrifugation, suggesting an association of the enzyme with the bacterial cell envelope. PMID- 8424690 TI - Retinoids inhibit the oxidative modification of protein kinase C induced by oxidant tumor promoters. AB - Recently we reported that oxidant tumor promoters can induce the oxidative modification of protein kinase C (PKC) resulting in either activation or inactivation of the kinase (R. Gopalakrishna and W. B. Anderson, Arch. Biochem. Biophys. 285, 382-387, 1991). Since retinoids previously have been shown to antagonize the actions of tumor promoters, studies were carried out to determine if retinoids can inhibit the oxidative modification of PKC induced by tumor promoters. Prior treatment of B16 melanoma cells or C6 glioma cells with all trans-retinoic acid (0.1 microM) for a short time period (15 to 60 min) followed by subsequent treatment with oxidants such as hydrogen peroxide resulted in a 30 to 70% decrease in the oxidative modification of PKC. This resulted in a decrease in oxidant-induced conversion of PKC from a Ca2+/lipid-dependent form (peak A) to a Ca2+/lipid-independent form (peak B). This retinoid-mediated protection also was observed with the reversible oxidative modification of PKC induced by m periodate treatment of intact cells. To understand whether this protection offered by retinoids was caused by a direct influence of retinoids on PKC, experiments were carried out using the purified enzyme. The results of experiments using isolated PKC suggested that retinoids can act directly to protect the regulatory domain of PKC from oxidative modification induced by oxidants. However, high (1-10 microM) concentrations of retinoids are necessary to elicit this protection of isolated PKC. In contrast, in experiments with intact cells, only low (submicromolar) concentrations of retinoids are required to protect PKC from oxidation. The differences noted in the retinoid concentrations required to protect PKC from oxidant modification in the test tube versus in the intact cell may be due to increased retention of retinoids in the cell membrane by partitioning, or to other indirect actions of retinoids in the intact cells to decrease cellular oxidations. These results suggest that some of the anti-tumor promoter actions of retinoids may be mediated, in part, by inhibiting the oxidative modification of protein kinase C induced by oxidant tumor promoters. PMID- 8424691 TI - Isozymes of lignin peroxidase and manganese(II) peroxidase from the white-rot basidiomycete Trametes versicolor. II. Partial sequences, peptide maps, and amino acid and carbohydrate compositions. AB - The basidiomycete Trametes versicolor, a white-rot fungus and potent degrader of lignin, produces multiple forms of extracellular peroxidases. Nine of these forms, six lignin peroxidases and three manganese(II) peroxidases, purified as described in the preceding paper, were characterized by amino-terminal sequencing, amino acid analyses, carbohydrate analyses, or peptide mapping. For two of the lignin peroxidase forms, tryptic peptides were isolated and sequenced to an extent corresponding to about 40 and 30%, respectively, of the primary structure. Eight of the nine peroxidases investigated were found to possess unique amino-terminal regions. A comparison of the sequences shows 57% of the residues to be identical, indicating a common ancestry for the lignin peroxidase and the manganese(II) peroxidase. The degree of identity among the five lignin peroxidases is about 80% and among the three manganese(II) peroxidases about 70%. Pairwise comparisons of the sequences disclosed that some of the lignin peroxidases are very closely related, either identical or differing only in a single amino acid residue of the thirty-five investigated. These close relationships are also supported by peptide mapping and by similarities in amino acid compositions. Tyr is absent in all isozymes. Lignin and manganese(II) peroxidases showed the presence of glucosamine and mannose in an amount corresponding to 3 to 6% of the molecular mass of the proteins. The carbohydrate compositions are compatible with the presence of 1, 2, and 3 sites of N glycosylation. The results obtained strongly suggest that the complexity in the peroxidase pattern displayed by the fungus (T. Johansson and P.O. Nyman, Arch. Biochem. Biophys. 300, 49-56, 1993) can largely be accounted for by a heterogeneity at the gene level, probably in the form of multiple structural genes. Two recently published genes from genomic clones of T. versicolor are identical in sequence to two of the lignin peroxidases characterized here. PMID- 8424692 TI - The development-associated increase in the hepatic levels of the intrinsic components of the chicken glycine cleavage system. AB - The hepatic glycine cleavage system in the chicken was examined at different stages of development to study regulation of its biosynthesis. Embryonic levels of polypeptide and mRNA for glycine decarboxylase, one of the three intrinsic components of this enzyme system, were approximately 10% of their adult levels and were rapidly increased following hatching. Those of H-protein, another intrinsic component, were somewhat higher and were more slowly increased throughout the course of development. The change in activities of the two components went with the increase in their polypeptide levels. Moreover, both relative levels of mRNAs for the two components and relative efficiencies of transcription of their genes were not constant during development. T-protein biosynthesis appeared to follow a course similar to that of H-protein. These observations imply that during development, the production of H-protein and T protein is regulated by similar mechanisms. Further, it is likely that the same mechanisms do not direct the regulatory processes in the biosynthesis of glycine decarboxylase and that embryo and adult livers possess different regulatory modes regarding the biosynthesis of this enzyme system. The shift to the adult mode appeared to occur at birth. The glycine cleavage activity that was three times higher than that in 12-day-old embryo was observed in adult liver mitochondria. PMID- 8424693 TI - Cytotoxicity and kinetic analysis of purified bovine serum amine oxidase in the presence of spermine in Chinese hamster ovary cells. AB - Bovine serum amine oxidase (BSAO, EC 1.4.3.6) catalyzes the oxidative deamination of polyamines giving rise to the corresponding aldehydes, ammonia and hydrogen peroxide (H2O2). This study demonstrates that amine oxidase (BSAO) purified from bovine serum and exogenous spermine caused cytotoxicity in Chinese hamster ovary (CHO) cells. Cytotoxicity occurred when cells were exposed to BSAO (0.0164-16.4 micrograms/ml) in the presence of spermine (1.9-340 microM). BSAO and spermine alone were not toxic at these concentrations. Cytotoxicity was dependent on the concentration of spermine and on the incubation time, and was also accelerated at 42 degrees C relative to 37 degrees C. Kinetic analysis of the enzymatic reaction, as a function of spermine concentration, showed Michaelis-Menten saturation kinetics. The apparent Vmax increased from 19.1 +/- 0.4 microM min-1 at 37 degrees C to 23.0 +/- 0.3 microM min-1 at 42 degrees C. The apparent Km decreased from 25.5 +/- 2.6 microM at 37 degrees C to 17.7 +/- 1.3 microM at 42 degrees C. Catalase inhibited cytotoxicity, suggesting that H2O2 was partially responsible for cytotoxicity. This work shows that the oxidation products of polyamines, rather than the polyamines themselves, are responsible for cytotoxicity in mammalian cells. The significance of this study is that amine oxidases could have therapeutic potential in cancer treatment regimens and a beneficial effect is likely when the enzyme is used together with clinical hyperthermia. PMID- 8424694 TI - In vivo fate of hemopexin and heme-hemopexin complexes in the rat. AB - The disposition in the rat of the plasma heme-binding protein hemopexin (Hx), as the native apoprotein and as its heme complex (HHx), has been studied using the residualizing protein label dilactitol-125I-tyramine (*I-DLT). The aim of this work was to identify the tissue sites of Hx uptake and catabolism, independent of heme binding, and to evaluate how heme loading affects Hx catabolism at these sites. *I-DLT-Hx had a circulating half-life of approximately 1.2 days and was recovered in degraded form in comparable amounts in visceral (liver, kidney, spleen) and peripheral (skin, muscle) tissues, indicating a generalized diffuse catabolism of the protein throughout the body. The plasma half-life of *I-DLT-Hx injected as a preformed heme-Hx complex was the same as that of the apoprotein; however, injection of the complex resulted in about a twofold increase in hepatic degradation of Hx. The lack of an effect of heme on overall catabolism of the preformed HHx complex was consistent with the approximately 1-h half-life of heme, injected as 14C-heme-Hx, in the circulation; however, as much as 20-fold more 14C-heme than Hx protein was recovered in liver from 14C-heme-Hx. The absolute amount of *I-DLT-Hx degraded in liver was significantly increased when heme was injected in excess of the heme binding capacity of circulating Hx, while 131I-DLT-albumin catabolism in liver was unaffected. Thus, depending on the physiological conditions studied, the data are consistent with a model in which, following hepatic uptake of heme from HHx, varying proportions of the protein are either returned to the circulation or degraded in the liver. PMID- 8424695 TI - Clinicopathologic factors relating to surgical margins for cutaneous melanoma. PMID- 8424696 TI - Liver resection under total vascular isolation. Variations on a theme. AB - Total vascular isolation (TVI) of the liver was employed during parenchymal transection in 16 patients undergoing hepatic resection for large tumors (mean diameter, 10.7 cm) located near hilar structures, hepatic veins, or the inferior vena cava (IVC). In 14 cases, TVI was achieved by clamping the suprahepatic and infrahepatic IVC and the porta hepatis, with or without aortic occlusion; in two, selective hepatic vein clamping was possible, obviating IVC occlusion. Procedures included standard and extended right and left lobectomies and caudate lobe resections. Concomitant resection and reconstruction of the portal vein (one case), IVC (one case), and bile duct (three cases) was required. Postoperative hepatic and renal failure did not occur. Mean intensive care unit and hospital stays were 2.8 +/- 1.9 and 12.5 +/- 5.2 days, respectively. There were two perioperative deaths. Total vascular isolation permits safe resection of large, critically located tumors that would otherwise present prohibitive operative risks. PMID- 8424697 TI - Melanoma recurrence after excision. Is a wide margin justified? AB - Through retrospective analysis of patient records for 187 patients with melanoma seen between 1975 and 1989, the aim of this study was to determine whether outcome varied according to degrees of surgical intervention in the primary treatment of stage I disease for thin, intermediate, and thick lesions. There were no significant differences in recurrence rate associated with an excision margin of 15 mm or less compared with wider excision margins; with initial excision compared with wider re-excision after excision biopsy; or for primary closure as compared with closure with a graft. There was, however, a significant difference in wound complication rate between wounds closed primarily (6%) and those closed by grafting (31%) (p < 0.01). The authors advocate the more conservative excision margin of 1.00 cm to 1.50 cm in the treatment of stage I melanoma with primary closure of the wound where possible. PMID- 8424698 TI - Sinistral portal hypertension. Splenectomy or expectant management. AB - Splenectomy has been considered the treatment of choice for patients with bleeding from sinistral portal hypertension (SPH) and varices, but is controversial for asymptomatic patients. To further define the role of splenectomy for SPH, the authors compared the clinical presentations and outcomes of 25 patients treated with splenectomy with those of 12 observed patients. Clinical features were similar except for transfusions administered (60% vs. 25%, p < 0.05), hemoglobin (9.8 +/- 2.2 g/dL vs. 12.5 +/- 2.1 g/dL, p < 0.05), and history of prior bleeding episodes (56% vs. 8%, p < 0.05), splenectomy versus no splenectomy, respectively. At 3 years, neither survival (78% vs. 64%, p = 1.0) nor new or recurrent bleeding (16% vs. 24%, p = 0.2) differed, splenectomy versus no splenectomy, respectively. The authors conclude that in the absence of prior bleeding episodes, anemia, or severe hemorrhage, observation of patients with SPH is justified. PMID- 8424699 TI - Cyst fluid analysis in the differential diagnosis of pancreatic cysts. A comparison of pseudocysts, serous cystadenomas, mucinous cystic neoplasms, and mucinous cystadenocarcinoma. AB - Pancreatic cystic lesions include inflammatory pseudocysts, benign serous tumors, and mucinous neoplasms, some of which are malignant. Clinical and radiologic indices are often inadequate to discriminate reliably among these possibilities. In an attempt to develop new preoperative diagnostic criteria to assist in decisions regarding therapy, the authors have performed cyst fluid analysis for tumor markers (carcinoembryonic antigen: CEA, CA 125, and CA 19.9), amylase content, amylase isoenzymes, relative viscosity, and cytology on 26 pancreatic cysts. The cases included nine pseudocysts, five serous cystadenomas, 4 mucinous cystic neoplasms, 7 mucinous cystadenocarcinomas, and one mucinous ductal adenocarcinoma with cystic degeneration. Carcinoembryonic antigen levels were high (> 367) in all benign and malignant mucinous cysts, but were low (< 23) in the pseudocysts and benign serous cystadenomas, an indication that CEA discriminates between mucinous and nonmucinous cysts (p < 0.0001). Values for CA 125 were high in all malignant cysts, low in pseudocysts, and variable in mucinous cystic neoplasms and serous cystadenomas. Levels of Ca 19.9 were nondiscriminatory. Cyst fluid amylase and lipase content were variable but were generally high in pseudocysts and low in cystic tumors. Amylase isoenzyme analysis was useful to differentiate pseudocysts from cystic tumors. Measurement of the relative viscosity in cyst fluid showed high (> serum viscosity) values in 89% of mucinous tumors and low values (< serum) in all pseudocysts and serous cystadenomas (p < 0.01). Cytologic analysis of cyst fluids was of limited value in differentiating pseudocysts from serous cystadenoma, but in seven of eight mucinous tumors provided useful diagnostic information and correctly classified three of five malignant tumors. The authors conclude that cyst fluid analysis can provide a preoperative classification of these diagnostically difficult lesions. The combination of viscosity, CEA, CA 125, and cytology can reliably distinguish malignant cystic tumors and potentially premalignant mucinous cystic neoplasms from pseudocysts and serous cystadenomas. Amylase content with isoenzyme analysis is useful to identify pseudocysts. PMID- 8424700 TI - Intestinal ischemia-reperfusion injury causes pulmonary endothelial cell ATP depletion. AB - Intestinal ischemia-reperfusion is a common clinical event associated with both clinical and experimental distant organ injury. In particular, the pulmonary microvasculature appears to be susceptible to injury resulting from systemic inflammatory mediator activation. This study was designed to evaluate the hypothesis that noncellular humoral factors associated with intestinal ischemia reperfusion result in pulmonary endothelial cell adenosine triphosphate (ATP) depletion. Male Sprague-Dawley rats had intestinal ischemia induced by microvascular clip occlusion of the superior mesenteric artery (SMA) for 120 minutes. Reperfusion resulted from superior mesenteric artery clip removal. After reperfusion for 0, 15, or 30 minutes, plasma samples were obtained from the portal vein. Monolayers of cultured rat pulmonary artery endothelial cells then were incubated with the plasma samples. Adenosine triphosphate levels were determined using a luciferin-luciferase assay. A 51Cr-release assay using labeled endothelial cells was performed under identical conditions to assess cytotoxicity. Potential mechanisms of ATP depletion were evaluated by analysis of cellular energy charge and assessment of microfilament architecture. Endothelial cell ATP levels decreased from 2.23 +/- 0.16 x 10(-11) moles/microgram DNA in sham preparations to 1.23 +/- 0.09 x 10(-11) moles/microgram DNA (p < 0.001) after 4 hours in plasma from animals undergoing 120 minutes of intestinal ischemia. For plasma obtained after 15 minutes of reperfusion, the decrease in cellular ATP concentration persisted (1.23 +/- 0.27 x 10(-11) moles/microgram DNA, p < 0.001 vs. sham). After 30 minutes' reperfusion, cellular ATP levels increased only slightly after the 4-hour incubation (1.39 +/- 0.26 x 10(-11) moles/microgram DNA, p < 0.005 vs. sham). No significant cytotoxic injury occurred in any group when compared with controls. Cellular energy charge was unchanged, and microfilament architecture was preserved. These data confirm the hypothesis that humoral factors, independent of the neutrophil, result in endothelial cell ATP depletion without metabolic inhibition or cell death. Depletion of energy stores by noncellular humoral factors may represent an early event that predisposes the cell to more severe injury by other mediators of the endogenous inflammatory response. PMID- 8424701 TI - Small bowel origin and calorie dependence of a signal for meal-induced jejunal absorption. AB - The ingestion of a meal stimulates the absorption of water and electrolytes from the small intestine independent of the cephalic or gastric phases of digestion. This study tested two hypotheses: (1) the jejunum is the origin of a postmeal proabsorptive signal and (2) the magnitude of the proabsorptive response is dependent on the caloric content of the meal stimulus. Twenty-five-centimeter proximal canine jejunal Thiry-Vella fistulas and feeding jejunostomies were constructed under general anesthesia. Jejunal absorption studies (n = 50) were performed by luminal perfusion of the Thiry-Vella fistula with 14C-polyethylene glycol (PEG) to calculate fluxes of water and electrolytes. Five groups were studied: (1) CONTROL: no meal, (2) 240 kcal oral meal, (3) 480 kcal oral meal, (4) 240 kcal jejunal meal, and (5) 480 kcal jejunal meal. Independent of the route of delivery (i.e., oral vs. jejunal), each meal stimulus significantly increased jejunal water and electrolyte absorption (p < 0.05). The magnitude of the proabsorptive response increased significantly as the calories delivered increased (p < 0.05). These data support the hypothesis that a proabsorptive signal responsible for meal-induced jejunal absorption originates from, or distal to the jejunum and suggest that intestinal chemoreceptors or osmoreceptors participate in the generation of the proabsorptive signal. PMID- 8424702 TI - Six-year results of a prospective, randomized trial of selective proximal vagotomy with and without pyloroplasty in the treatment of duodenal, pyloric, and prepyloric ulcers. AB - In a consecutive series of patients with uncomplicated prepyloric, pyloric, or duodenal ulcer, 39 patients were randomly allocated to selective proximal vagotomy with pyloroplasty, and 40 patients to selective proximal vagotomy alone with no operative mortality. Before surgery, all patients had undergone H2 receptor antagonist treatment. No patient was lost for follow-up. At an average follow-up of 6 years, recurrent ulcer was recorded in 15% and 20%, respectively, after selective proximal vagotomy with and without pyloroplasty. Three of 14 recurrent ulcers were asymptomatic. Epigastric pain with or without ulcer was significantly less common after selective proximal vagotomy with (13%) than without pyloroplasty (40%). Mild diarrhea or mild dumping was recorded in a few patients. The overall results were very good or good (Visick I or II) in 77% and 55% (significant difference) after vagotomy with and without pyloroplasty, respectively, and in 82% and 58%, if asymptomatic ulcers were graded as Visick I or II results. Of the 27 patients with Visick III or IV results, three patients needed no treatment (asymptomatic ulcers), and 10 patients had no symptoms during medical treatment. Two patients with vagotomy and pyloroplasty and nine with vagotomy alone were reoperated. There were no deaths, and the results were graded as Visick I or II in 10 patients and as Visick III in one patient. The authors conclude that selective proximal vagotomy with pyloroplasty is superior to vagotomy alone for the treatment of prepyloric-pyloric and duodenal ulcer. Recurrent ulcer after vagotomy has a benign course and responds well to ranitidine treatment. PMID- 8424703 TI - Endorectal ultrasound for control of preoperative radiotherapy of rectal cancer. AB - Endorectal ultrasound (EUS) is known to be a reliable method for preoperative staging of rectal tumors. In this study, EUS was used to select patients with rectal cancer suitable for preoperative radiation therapy. By performing EUS before and after radiation, the aim of the study was to evaluate the role of EUS in monitoring the effects of preoperative radiation therapy. In 17 patients with large T3 or T4 rectal tumors, a complete staging by EUS was done before and after radiation therapy. Beside a shrinkage of the tumor, there was a change of echopattern to more hyperechoic gray levels to be observed in the irradiated tumor. The rectal wall lost its normal architecture, and lymph nodes disappeared or changed their echopattern from echopoor to echorich. There was no down-staging of a tumor seen by EUS. Complete preoperative staging was correct in 13 of 17 patients because of endosonographic examination before and after preoperative radiation therapy. New interpretation criteria are given for evaluation of patients with rectal cancer treated by radiation therapy. PMID- 8424704 TI - Lymph node metastasis from soft tissue sarcoma in adults. Analysis of data from a prospective database of 1772 sarcoma patients. AB - To examine the natural history of lymph node metastasis from sarcomas and the utility of therapeutic lymphadenectomy, clinical histories of all adult patients identified by a prospective sarcoma database for the 10-year period July 1982 to July 1991 were examined. Of the 1772 sarcoma patients, 46 (2.6%) were identified with lymph node metastasis. Median follow-up of all patients from diagnosis of lymph node metastasis was 12.9 months (range, 0 to 100 months). Median survival for nonsurvivors was 12.7 months (range, 0 to 40.7). The tumor types with the highest incidence of lymph node metastasis are angiosarcoma (5/37 total cases; 13.5%), embryonal rhabdomyosarcoma (ERMS) (12/88 total cases; 13.6%), and epithelioid sarcoma (2/12 total cases; 16.7%). Lymph node metastasis from visceral primary (p = 0.004) and malignant fibrous histiocytomas (p = 0.006) were associated with particularly poor prognosis. Thirty-one patients underwent radical, therapeutic lymphadenectomy with curative intent, whereas 15 patients had less than curative procedures, in most cases biopsy only. Patients not treated with radical lymphadenectomy had a median survival of 4.3 months (range, 1 to 32) whereas radical lymphadenectomy was associated with a 16.3 month median survival and the only long-term survivors (46% 5-year survival by Kaplan-Meier). The authors conclude that lymph node metastases from sarcoma are rare in adults, but vigilance is warranted, especially in angiosarcoma, ERMS, and epithelioid subtypes. Radical lymphadenectomy is appropriate treatment for isolated metastasis to regional lymph nodes and may provide long-term survival. PMID- 8424705 TI - Murphy's Button revisited. Clinical experience with the biofragmentable anastomotic ring. AB - Use of the biofragmentable anastomosis ring (BAR) was attempted in 33 patients at two New York City institutions and employed in 31 instances. Anastomoses performed were end-to-end enterocolic (n = 15), colocolic (n = 15), and side-to side colocolic (n = 1). Patients ranged in age from 27 to 86 years, with the following diagnoses: primary colon cancer, 15; sessile adenoma, four; colostomy, five; diverticulosis, two; metastatic cancer with obstruction, multiple polyposis, perforated appendiceal mass, malignant carcinoid of appendix, intussuscepting right colon mass, one each. In two instances use of the device was aborted because of concern with the blood supply to the bowel wall in one and tissue edema in another. The average duration of postoperative ileus was 4.7 days. Two patients were subsequently treated for small bowel obstruction thought unrelated to use of the anastomotic device. There were no deaths and no evidence of stricture. PMID- 8424706 TI - Surgical techniques and innovations in living related liver transplantation. AB - The authors successfully performed a series of 33 living related liver transplantations (LRLT) on children (15 males and 18 females, ranging from 7 months to 15 years of age) from June 1990 to May 1992, with the informed consent of their parents and the approval of the Ethics Committee of Kyoto University. Before operation, six of the children required intensive care, another 14 were hospitalized, and 13 were homebound. Donors (12 paternal and 21 maternal) were selected solely from the parents of the recipients on the basis of ABO blood group and graft/recipient size matching determined by computed tomography scanning. Procurement of graft was performed using ultrasonic aspirator and bipolar electrocautery without blood vessel clamping and without graft manipulation. All donors subsequently had normal liver function and returned to normal life. The left lateral segment (16 cases), left lobe (16 cases), or right lobe (one case) were used as grafts. The partial liver graft was transplanted into the recipient who underwent total hepatectomy with preservation of the inferior vena cava using a vascular side clamp. Twenty-seven of 33 recipients are alive and well with the original graft and have normal liver function. The patient survival rate was 89% (24/27) in elective cases and 50% (3/6) in emergent cases. The other six recipients had functioning grafts but died of extrahepatic complications. Complications of the graft were minimal in all cases. Hepatic vein stenosis, which occurred three times in two cases, was successfully treated by balloon dilatation. In cases with sclerotic portal vein, the authors anastomosed the portal vein of the graft to the confluence of the splenic vein and the superior mesenteric vein without a vascular graft, after experiencing a case of vascular graft thrombosis. After hepatic artery thrombosis occurred in one of the initial seven recipients whose arterial anastomosis was done with surgical loupe, microsurgery was introduced for hepatic artery reconstruction. There has been no occurrence of thrombosis since then. The current results with LRLT suggested that the meticulous management of surgical factors at each stage of the LRLT procedure is crucial for successful outcome. Living related liver transplantation is a promising option for resolving the graft shortage in pediatric liver transplantation and may be regarded as an independent modality to supplement cadaver donation. PMID- 8424707 TI - Alcohol use and risk of posterior subcapsular opacities. AB - We conducted a follow-up study of surgical cases of posterior subcapsular cataracts and their controls to evaluate the possible association of alcohol intake and posterior subcapsular opacities. Two hundred thirty-eight cases and controls were interviewed. Current alcohol intake and usual and maximum weekly consumption ever were assessed. In this population, 57% of the cases and 56% of the controls were nondrinkers, 22% of the cases and 36% of the controls had an average of seven or fewer drinks per week, and 17% of the cases and 8% of the controls had more than seven drinks per week. A matched pair analysis controlling for other known risk factors showed an increased risk associated with heavy alcohol use. Heavy drinkers were more likely to be cases than were nondrinkers (odds ratio, 4.6; P < .05), and light drinkers were not at an increased risk. This result suggests that heavy alcohol consumption may increase the risk of posterior subcapsular cataract. PMID- 8424708 TI - Alcohol use and lens opacities in the Beaver Dam Eye Study. AB - The relationship between alcohol use and lens opacities was examined in a large (N = 4926) population-based study of adults aged 43 to 86 years in Beaver Dam, Wis. These data were collected from 1988 to 1990. Alcohol history was determined by a standardized questionnaire. Prevalence and severity of cataract were determined by masked grading of photographs obtained using a slit-lamp camera and retroillumination. A history of heavy drinking was related to more severe nuclear sclerotic, cortical, and posterior subcapsular opacities (odds ratios, 1.34, 1.38, and 1.57, respectively). These relationships remained after adjusting for other risk factors such as smoking. Moderate liquor consumption was associated with less severe nuclear sclerosis (odds ratio, 0.81). Participants who drank wine had less severe nuclear sclerosis (odds ratio, 0.84) and cortical opacities (odds ratio, 0.84) than those who did not. Increased consumption of beer was related to increased risk of cortical opacities. PMID- 8424709 TI - Pharmacokinetics of newer cephalosporins after subconjunctival and intravitreal injection in rabbits. AB - Pharmacologic considerations suggest that third-generation cephalosporins might penetrate the vitreous humor better after periocular injection and might be eliminated less readily after intravitreous injection than older agents. We studied the sodium salts of ceftizoxime, ceftriaxone, and ceftazidime, and of an investigational cephalosporin, cefepime, in rabbits. After a single subconjunctival injection in animals with normal eyes, vitreous levels ranged from 3 to 13 mg/L. After five subconjunctival injections in rabbits with infected eyes, vitreous concentrations ranged from 12 to 34 mg/L. These concentrations are not appreciably greater than those found with older beta-lactams. The vitreous half-life of the four drugs after intravitreous injection varied from 5.7 to 20 hours in rabbits with uninflamed eyes and from 9.4 to 21.5 hours in rabbits with infected eyes. Except for ceftizoxime, the half-lives were substantially longer than those for older beta-lactams and suggest predominantly anterior route elimination. Vitreous penetration of these new agents after subconjunctival injection does not appear to be sufficient to overcome the need for intravitreous injections in the treatment of endophthalmitis. However, the longer vitreous half lives of some of the newer agents may be useful if the drugs are to be given intravitreally. PMID- 8424710 TI - Light-induced deposits in Bruch's membrane of protoporphyric mice. AB - Photosensitization of choriocapillary endothelium with blood-borne photosensitizers, such as protoporphyrin IX, has been proposed as a mechanism for the choriocapillary sclerosis and Bruch's membrane deposits seen in age-related macular degeneration. Utilizing a mouse model of protoporphyria with approximately a 10-fold increase in protoporphyrin IX level and exposure to blue light (14 microW/cm2; bandwidth, 390 to 430 nm), a time- and light-dependent increase in choriocapillary and sub-retinal pigment epithelium basal laminar-like deposits could be demonstrated at 7 months by transmission electron microscopy. Thickening of the choriocapillary endothelial basement membrane with a homogeneous electron-dense material was first noted in protoporphyric mice exposed to blue light for 13 weeks. At 28 weeks the experimental animals exhibited a thick band of homogeneous deposits at the level of the choriocapillary basement membrane and electron-dense fibrillogranular deposits of varying sizes along the inner aspect of Bruch's membrane, with fibrils measuring up to 16 nm, with a periodicity of 13 nm. These deposits contributed to an overall thickening of Bruch's membrane with narrowing of the choriocapillaris. The morphologic appearance and localization of these deposits within Bruch's membrane of this animal model are similar to previously described deposits noted in the aging Bruch's membrane prior to the development of age-related macular degeneration. PMID- 8424711 TI - Eosinophilic granuloma. PMID- 8424712 TI - Corneal toxicity from acidic vancomycin solution. PMID- 8424713 TI - Extracapsular cataract extraction in developing countries. PMID- 8424714 TI - Serous detachment after high-dose intravitreal dexamethasone: toxic or osmotic? PMID- 8424715 TI - Lymphocytic infiltration of the conjunctiva and the salivary gland in Sjogren's syndrome. PMID- 8424716 TI - Tissue analysis from two patients with premacular hole lesions. PMID- 8424717 TI - Immune-related disease and normal-tension glaucoma. PMID- 8424718 TI - Immune-related disease and normal-tension glaucoma. PMID- 8424719 TI - Pseudomonas aeruginosa iatrogenic blepharoconjunctivitis. PMID- 8424720 TI - Reversible unilateral cataract in a patient without overt diabetes. PMID- 8424721 TI - Bilateral extensive persistent pupillary membranes treated with the neodymium-YAG laser. PMID- 8424722 TI - National health care spending. Where do the dollars go? PMID- 8424723 TI - The Resource-Based Relative Value Scale. Methods, results, and impacts for ophthalmology. AB - In January 1992, the Health Care Financing Administration implemented sweeping legislation that reformed the way Medicare pays for physicians' services. The cornerstone of the reform consists of a new fee schedule based on the Resource Based Relative Value Scale. This article summarizes the methods and data used to derive the scale for ophthalmology. The results and impacts of the new Medicare payment rates for ophthalmology are also assessed. Using our methods and assumptions, ophthalmologists stand to lose 16% of their Medicare revenues under a fully implemented relative value-based fee schedule. Overall, the fees for performing evaluation and management services will increase, while those for most surgical procedures and diagnostic tests will decrease. Physicians' practice decisions and medical students' specialty choices could be affected. Ophthalmologists who perform work-intensive surgical procedures and spend the majority of their time in the operating room will continue to earn much higher incomes than those who do not. PMID- 8424724 TI - Ophthalmology and the Resource-Based Relative Value Fee Scale. AB - The Medicare Resource-Based Relative Value Scale for ophthalmology has significantly reduced the level of reimbursement for surgical fees and only minimally increased evaluation and management fees. Some observers have felt that the methods for determining fees were flawed, and, generally, practitioners have been concerned about a potential loss of income. While reimbursement for individual services is being cut, projections through 1996 indicate that ophthalmology, as a specialty, will receive 55% more funding due to historical trends and increasing ranks of providers. This will translate into a more moderate global reduction in revenue of approximately 11%. The possible implications of the Resource-Based Relative Value Scale include a concentration of ophthalmic surgery into fewer practices, which may be able to distribute medical liability costs over a larger number of procedures. To counter the constraints of fee limits, individual physicians will probably seek to enhance their net income by greater use of paraprofessional personnel, the acquisition of new technologies, and the application of improved management skills. PMID- 8424725 TI - Comparison of acuity, contrast sensitivity, and disability glare before and after cataract surgery. AB - We assessed vision before and after uncomplicated extracapsular cataract extraction and intraocular lens implantation in 72 symptomatic patients with acuity equal to or better than 20/80 and no other ocular abnormality. Contrast sensitivity was measured with the Pelli-Robson Letter Chart (Metropia Ltd, Cambridge, England) and disability glare was measured under daytime conditions with the Brightness Acuity Tester (Mentor O&O Inc, Norwell, Mass) and under nighttime conditions with a computer-controlled video display. Prior to surgery there was significant disability glare that was not correlated with acuity. There was also a loss in contrast sensitivity that was moderately correlated with acuity (r = -.43; P < .001). Following surgery, most patients' scores returned to normal on all tests. Improvement in disability glare and contrast sensitivity was independent of improvement in acuity. Furthermore, patients with the poorest preoperative vision were as likely to regain normal function after surgery as those with the best preoperative vision. PMID- 8424726 TI - Progression of disc and field damage in early glaucoma. AB - To assess the temporal relationship between visual field progression and optic disc deterioration in early glaucoma, we studied 15 patients with unilateral visual field loss from primary open angle glaucoma. Planimetric optic disc measurements were compared with automated static threshold perimetry during a mean follow-up of 6.1 years. Eight (53%) of 15 eyes with an initially normal visual field showed progression of the disc; six of these eyes did not develop field abnormalities. The mean rates of rim-area loss were 1.7%/y in eyes with initially normal fields and 2.1%/y in eyes with initial field loss. The mean rate of visual field deterioration (change in corrected loss variance) was lower in the eyes with an initially normal field (0.3 dB2/y) than in eyes with initial field loss (3.6 dB2/y; P = .016). This longitudinal study documents progressive disc damage prior to field loss in early glaucoma. PMID- 8424727 TI - Optic nerve hypoplasia. Clinical significance of associated central nervous system abnormalities on magnetic resonance imaging. AB - Magnetic resonance imaging demonstrated coexistent central nervous system abnormalities in 30 of 40 patients with optic nerve hypoplasia. Based on their associated neuroradiological findings, these patients were placed into one of five categories: group 1, isolated optic nerve hypoplasia; group 2, absence of the septum pellucidum; group 3, posterior pituitary ectopia; group 4, hemispheric migration anomalies; and group 5, intrauterine/perinatal hemispheric injury. Posterior pituitary ectopia (group 3) and cerebral hemispheric abnormalities (groups 4 and 5) were found to be highly predictive of pituitary hormone deficiency and neurodevelopmental deficits, respectively. Isolated absence of the septum pellucidum (group 2) was associated with normal neurodevelopmental and endocrinologic function. Thinning or agenesis of the corpus callosum was predictive of neurodevelopmental problems only by virtue of its frequent association with cerebral hemispheric abnormalities. Magnetic resonance imaging can now be used to provide specific prognostic information regarding the likelihood of neurodevelopmental deficits and pituitary hormone deficiency in infants with optic nerve hypoplasia. The prevailing notion of septo-optic dysplasia as a distinct nosologic entity should be reconsidered. PMID- 8424728 TI - Factors influencing response to strabismus surgery. AB - Based on analyses in a series of 116 patients, we found that the response to strabismus surgery (degrees of change of ocular alignment per millimeter of rectus recession) correlated significantly with the preoperative deviation for esotropic and exotropic patients. The prediction of response to strabismus surgery was not improved significantly with the inclusion of axial length, age, and/or preoperative refractive error beyond the prediction provided with use of only the preoperative deviation, even though we have previously suggested that the response to strabismus surgery should be related to axial length. We believed that larger eyes should have a smaller response for the same number of millimeters of surgery than smaller eyes. We now believe that although the response to strabismus surgery does correlate significantly and inversely with axial length, this correlation may not be clinically important given the much stronger influence of preoperative deviation. PMID- 8424729 TI - Predictive factors for endothelial cell loss after penetrating keratoplasty. AB - To assess the influence of donor age, donor endothelial cell density, recipient age, and diagnosis on loss of endothelial cell density after penetrating keratoplasty, we evaluated endothelial cell density data from specular endothelial images collected during the follow-up penetrating keratoplasty surgeries performed from 1980 through 1985 at one center. Univariate and multivariate analyses of information from 265 grafts showed consistent and statistically significant associations between 1-year postoperative loss of endothelial cell density and donor age, donor endothelial cell density, and recipient age. Corneas obtained from older donors, corneas with higher donor endothelial cell density, and corneas transplanted to older recipients demonstrated greater percentage of loss of endothelial cell density 1 year after surgery. These three factors accounted for 17% of the total variance in percentage of loss of endothelial cell density 1 year after surgery. PMID- 8424730 TI - Optic nerve sheath fenestration for treatment of progressive ischemic optic neuropathy. Results in 26 patients. AB - Optic nerve sheath fenestration was performed in 26 eyes for treatment of the progressive type of common (nonarteritic) anterior ischemic optic neuropathy. During a mean follow-up period of 21 weeks (range, 6 to 52 weeks), results were as follows: visual acuity increased by two or more lines on the Snellen chart in 7 eyes; visual acuity decreased by two or more lines in four eyes; some regression of visual field defects occurred in six eyes, including two eyes in which acuity also improved. These results, attributed to surgical decompression, do not exceed the spontaneous recovery rates reported in the literature pertaining to nonarteritic anterior ischemic optic neuropathy and fail to substantiate the sanguine visual outcome in recently reported series of patients undergoing optic nerve sheath decompression. PMID- 8424731 TI - Ultrastructural features of surgically excised subretinal neovascular membranes in the ocular histoplasmosis syndrome. AB - We evaluated the ultrastructural features of seven surgically excised submacular neovascular membranes from seven patients with the ocular histoplasmosis syndrome. Excised neovascular membranes were composed of fibrovascular tissue interposed between Bruch's membrane and the retinal pigment epithelium. Cellular components present in the membranes included, in decreasing order of frequency, retinal pigment epithelium, vascular endothelium, photoreceptor cells, macrophages, erythrocytes, ghost erythrocytes, fibrocytes, myofibroblasts, glial cells, smooth-muscle cells, and lymphocytes. Extracellular constituents of the neovascular membranes included 20 to 25-nm collagen fibrils, fibrin, 10-nm collagen fibrils, and fragments of Bruch's membrane and choroid. Our findings are consistent with the concept that subretinal neovascular membranes in the ocular histoplasmosis syndrome represent a nonspecific healing response to a local stimulus or injury. PMID- 8424732 TI - Randomized clinical trials on medical treatment of glaucoma. Are they appropriate to guide clinical practice? AB - A systematic quantitative and qualitative overview of published randomized clinical trials was undertaken to assess the yield of medical treatment on the outcome of patients with primary open angle glaucoma. Reports of 102 randomized clinical trials were published between 1975 and 1991, totalling about 5000 patients. Only 16% (16/102) of the trials were, however, properly designed (ie, comparing an active treatment with a placebo-treated or untreated control group) to answer the question of whether any medical treatment can effectively cure patients with primary open angle glaucoma. Pooled analysis showed a moderate yet statistically significant reduction in mean intraocular pressure (-4.9 mm Hg; 95% confidence interval [CI], -7.3 to -2.5 mm Hg); however, data on long-term visual field changes were available in only three randomized clinical trials, and their statistical combination failed to show a significant protective effect of active treatment (odds ratio, 0.75; 95% CI, 0.42 to 1.35). All of the remaining 86 randomized clinical trials looked at the effectiveness of one drug vs another in lowering intraocular pressure and were thus of no use in the overview. Practicing ophthalmologists should be aware that the effectiveness of pressure-lowering agents in the treatment of primary open angle glaucoma is still to be determined and that the vast majority of published trials are not appropriate to guide clinical practice. It is urgent to plan trials with end-point and follow-up duration that is fully relevant for the health of patients. PMID- 8424734 TI - Prospective assessment of stereoscopic visual status and USAF pilot training attrition. AB - The role of stereopsis (i.e., the use of binocular cues for depth perception) in military aviation is undetermined. Pilots possessing adequate near stereopsis but having deficient distant stereopsis are thought to have microtropias. Historical reviews of microtropia and research concerning the role of depth perception in military aviation are described. A historical prospective study of student pilots entering U.S. Air Force Undergraduate Pilot Training (UPT) from Oct 1990 through Sep 1991 (FY 90-91) compares UPT attrition rates according to their preselection stereoscopic status (microtropia vs. normal). Univariate and multiple logistic regression analyses do not show significant differences in attrition rates between the two groups, implying that distant stereopsis is not critical to successful completion of UPT. The U.S. Air Force decided in Oct 91 to eliminate near stereoscopic vision screening while retaining distant stereoacuity testing as a criterion for candidates to qualify medically for UPT. Valid rationale for this decision includes simplified and uniform administration of stereoacuity testing, minimizing spurious results, the continued validity of stereopsis testing as a cross-check of other areas of visual function, the uncertain role of stereopsis in critical areas of flight operations, and the large applicant pool competing for a limited number of pilot training positions. PMID- 8424733 TI - Effects of scopolamine on autonomic profiles underlying motion sickness susceptibility. AB - The purpose of this study was to examine the effects of scopolamine on the physiological patterns occurring prior to and during motion sickness stimulation. In addition, the use of physiological profiles in the prediction of motion sickness was evaluated. Sixty subjects ingested either 0.6 mg scopolamine, 2.5 mg methscopolamine, or a placebo. Heart rate (HR), respiratory sinus arrhythmia (an index of vagal tone), and electrogastrograms were measured prior to and during the exposure to a rotating optokinetic drum. Compared to the other groups, the scopolamine group reported fewer motion sickness symptoms, and displayed lower HR, higher vagal tone, enhanced normal gastric myoelectric activity, and depressed gastric dysrhythmias before and during motion sickness induction. Distinct physiological profiles prior to drum rotation could reliably differentiate individuals who would develop gastric discomfort from those who would not. Symptom-free subjects were characterized by high levels of vagal tone and low HR across conditions, and by maintaining normal (3 cpm) electrogastrographic activity during drum rotation. It was concluded that scopolamine offered motion sickness protection by initiating a pattern of increased vagal tone and gastric myoelectric stability. PMID- 8424735 TI - Safety concerns as a factor in pilot desire to change aircraft. AB - An anonymous and voluntary questionnaire was completed by 461 (73.4%) Coast Guard aviators at 26 participating Air Stations. Most (92%) pilots felt that helicopters had more dangerous missions and were less safe than fixed wing aircraft (FXAC). Two thirds (204) of helicopter and 19.3% (29) of FXAC pilots stated a desire to change to a different aircraft category. Only 1 (3.6%) of the FXAC pilots but 81 (39.8%) of the helicopter pilots stated their wish for aircraft change was influenced by a desire to operate aircraft they perceived as safer. Pilots who desired change in aircraft category were more senior, more experienced, and felt they were currently engaging in dangerous flight operations. In conclusion this study found that: 1) Coast Guard pilots consider helicopters less safe than FXAC; 2) a substantial proportion of helicopter pilots would like to change to FXAC; 3) safety concerns can be a common factor in pilots' desires to change aircraft. PMID- 8424736 TI - Spectral analysis of the electroencephalographic response to motion sickness. AB - Ten subjects participated in a laboratory experiment using cross-coupled angular stimulation to induce motion sickness symptoms. A 14-channel montage using subdermal electrodes was employed to record the electroencephalogram during a pre Coriolis stimulation baseline through to imminent emesis. Spectral analyses of the EEG were performed upon the recorded data and individual band energies were quantified to attempt to characterize the cortical electrical response to motion sickness. Power spectral analysis was performed upon the temporo-frontal signals through the entire period over the delta, theta, and alpha EEG bands. The power in each of these bands was integrated and the baseline periods compared with that during frank sickness. Mean power spectral energy in the delta band during frank sickness increased by a factor of 13.7 over baseline. Mean theta band energy increased by a factor of 2.2. Mean alpha band energy was not significantly different. EEG power spectral levels in the delta and theta bands increased along with the level of motion sickness symptoms. These changes, particularly those in the delta band, suggest that intense low frequency oscillatory stimulation is being diffusely projected about the central nervous system. These EEG changes, similar to those sometimes seen in partial seizures, and the similarity of the symptom/sign complex in the two disorders, provide evidence that the pathophysiology and electrophysiology of motion sickness may be a variant of seizure activity. PMID- 8424737 TI - Effects of simulated high altitude exposure on long-latency event-related brain potentials and performance. AB - The N100, P200, N200 and P300 components of the auditory event-related potential were recorded from 10 male subjects at 0900, 1600, and 1830 hours at sea level and again following a rapid ascent to simulated 4300 m altitude. Amplitude and latency of components, ear oximetry, and concurrent performance measures (reaction time and counting errors) were assessed. Amplitude of P300 decreased, while P300 latency and reaction time increased, following ascent to altitude. However, the time course of altitude effects differed for amplitude versus latency. Components N100, P200, N200, and counting errors were unaffected by altitude. The results indicate that central measures of cognitive capacities are differentially sensitive to high altitude. The time course of altitude effects on P300 amplitude versus P300 latency suggests that the two measures reflect different aspects of a response to hypobaric hypoxia exposure. PMID- 8424738 TI - Operation Everest II: gas tensions in expired air and arterial blood at extreme altitude. AB - Measurements in alveolar gas have suggested extreme hypocapnia and alkalosis on the summit of Mt. Everest. However, tensions in both expired gas and arterial blood have not been reported for the summit of Mt. Everest (PIO2 = 43 mm Hg). To approach the problem, we examined alveolar (and end-tidal) and arterial gas tensions in 6 healthy men who completed a 40-d chamber study to the simulated "summit," with 20 d above 6,400 m and 9 d above 8,000 m. In 27 simultaneous samples of alveolar air and arterial blood for inspired oxygen tensions ranging from PIO2 of 55 mm Hg (7,380 m) to 43 mm Hg, the mean alveolar-arterial difference was negligible for PO2 (-0.8 +/- 2.4 (S.D.) mm Hg) and PCO2 (0.5 +/- 1.4 mm Hg). For all 6 subjects at the summit, PACO2 was 12.0 +/- 1.8 and PACO2 was 11.4 +/- 1.6 mm Hg, and for the two with the lowest values, alveolar and arterial PCO2, respectively, were 9.5 and 9.8 mm Hg. Arterial pH averaged 7.53 units. We conclude that while acclimatization to severe hypoxia results in extreme hypocapnia, alkalosis is more moderate than previously reported. Alveolar gas tensions reflect well the values obtained in arterial blood. PMID- 8424739 TI - Intracardiac hemodynamics in man during short periods of head-down and head-up tilt. AB - The intracardiac hemodynamic responses to short periods of 70 degrees head-down and head-up tilts were studied in 12 normal male subjects, ages 19-24 years. Echo Doppler techniques were used to measure the transmitral and transaortic flow velocities as well as cardiac index, and to evaluate the peripheral impedance. Head-down tilt (HDT) rapidly induced an increase (9.7%, p < 0.05) in the early passive filling of the left ventricle (ME peak of the transmitral flow velocity curve) and in transaortic flow velocity (8%, p < 0.05), as well as in cardiac output (6%, p < 0.05). In spite of a peripheral vasodilation, the blood pressure increased (7%, p < 0.05 for the systolic; 15%, p < 0.01 for the diastolic) and remained at a high level for the 5 min of the experiment. Head-up tilt (HUT) induced inverse responses; i.e., a large initial decrease in the transmitral ( 15%, p < 0.05) and transaortic (-16%, p < 0.001) flows. The shape of the arterial peripheral flow indicated an increased vascular impedance. After a short drop, the blood pressure rapidly recovered a level statistically close to that of the pretest. In both cases, tachycardia occurred. We conclude that, in man, the cardiac responses to the changes in posture appear to be related more to the passive changes in ventricular filling due to the blood shift than to the nervous regulation by the arterial baroreflexes, whereas these reflexes mainly act in the control of the vascular impedance. PMID- 8424740 TI - Response of the circadian system to 6 degrees head-down tilt bed rest. AB - The influence of simulated microgravity and reduced zeitgebers on the circadian system was examined in eight subjects. The 12-d study included a 3-d period for baseline measurements, 7 d of 6 degrees head-down bed rest, and 2 d for recovery. The sleep-wake cycle was kept constant during the study. The state of the circadian system was assessed from continuous measurements of temperature and ECG, and collections of urine at 3-h intervals for the determination of hormone and electrolyte excretions. Results indicate that only minor changes occurred in rhythm acrophases, whereas daily means for several physiological functions and their circadian amplitudes were significantly altered during the bed-rest phase. These studies using head-down tilt confirm previous findings with horizontal bed rest and indicate that rhythm disturbances can occur without change in the sleep wake cycle. To the degree that bed-rest studies simulate manned spaceflights, results indicate that variations in circadian cycles can occur even during short term missions and must be taken into account when interpreting subsequently obtained physiological data. PMID- 8424741 TI - Flight helmet weight, +Gz forces, and neck muscle strain. AB - The aim of this study was to investigate the effect of flight helmet weight on cervical erector spinae muscle strain under high +Gz forces. Two helmets of different weight were compared by recording inflight neck muscle activities with a portable surface-integrated EMG (IEMG) device. The obtained IEMG activities were normalized by comparing them with activities representing maximal voluntary contraction (MVC) of the muscles. Two test pilots carried out a total of 16 flights consisting of a series of different maneuvers. The results indicate that a lighter flight helmet may--at least in some pilots--cause less strain on neck structures than a heavier one. The effect of helmet weight was readily apparent only under high +Gz forces; changing from a heavier to a lighter helmet reduced the mean muscular strain from 9.5 to 8.8% and from 20.2 to 17.1% of the MVC under +4.0 and +7.0 Gz, respectively. Thus, some, but not all, acute inflight neck pain and related problems might be avoided by using lighter flight helmets. PMID- 8424742 TI - Animal surgery in microgravity. AB - The first surgical procedure in microgravity using an animal model is described. The investigation resulted in a realistic evaluation of prototype hardware and procedures that could be applied to a surgical support system on Space Station Freedom. This was especially true for the issue of the management of surgical bleeding, which was observed and studied in microgravity for the first time. PMID- 8424743 TI - Human factors in crashes of commuter airplanes. AB - Pilots and frequent travelers on commuter aircraft are exposed to higher risks of death or injury than those on major air carriers. To provide a better understanding of the circumstances of crashes of scheduled commuter airplanes, National Transportation Safety Board data were analyzed for all cases of death, serious injury, or major damage involving commuter airplanes during 1983-88, when 172 people were killed and 207 injured in 118 events. Three-fourths of cases involved inadequate pilot performance, notably poor handling of emergencies and improper instrument flying procedures. Pilot errors occurred disproportionately in bad weather, which played a role in 30% of crashes. Aircraft malfunctions were involved in 42% of crashes. Certain airplanes were overinvolved in gear-up landings or in crashes due to fuel mismanagement. Greater priority should be given to applying known preventive measures to the problem of commuter crashes. PMID- 8424744 TI - The physiological consequences of simulated helicopter flight in NBC protective equipment. AB - The physiological effects of wearing U.S. Army aviator nuclear-biological chemical (NBC) individual protective equipment (IPE) were evaluated in the USAARL UH-60 research flight simulator. There were 16 male aviators who flew the simulator in 4 test conditions: standard flight suit and cool cockpit, standard flight suit and hot cockpit, NBC IPE and cool cockpit, NBC IPE and hot cockpit. The cool condition was a WBGT of 17.9 degrees C, the hot 30.6 degrees C. Rectal temperature, mean skin temperature, and heart rate were monitored and showed significant increases for the NBC hot condition compared with the other three. Seven subjects failed to complete the sortie in the NBC hot condition, with a mean survival time of 298 min. All subjects flew for the target 6 h in the other conditions. PMID- 8424745 TI - Individual differences and subgroups within populations: the shopping bag approach. AB - The aerospace medical research community needs to consider the individual as something other than a statistical entity. The cumulative effects of performance enhancers that are collectively ignored, secondary to statistical analysis of populations, can be significant for individuals. By considering the individual, all aspects of whatever makes humans unique need to be integrated into research. One suggested remedy to the problem of subordination of the individual to the population mean/standard deviation is the use of a "shopping bag" approach. In this approach, each individual may select those performance enhancers that work best for him or her (based on controlled studies). Acceleration protection devices are used as an example. The impact of this philosophy can be readily seen in human factor design strategy as well as in the interpretation of human research data. PMID- 8424746 TI - You're the flight surgeon. Abdominal pain that has lasted 3 days. PMID- 8424747 TI - Extent of the effect air travel has had on cases of cholera reported in the United States. PMID- 8424748 TI - Hazard potential of ejection with canopy fragmentation. AB - The hazard potential of ejection with canopy fragmentation was evaluated by horizontal sled tests. A series of 14 ejections of tandem seats with 1st or 95th percentile anthropometric dummies were performed at sled speeds of 0, 150, 350, and 600 knots. Canopy-mounted miniature detonating cords (MDC) were fired in nine and not fired in five ejections. Maximal impact loads on each dummy's neck, knees, and shoulders were measured, and noise levels inside and outside the helmet recorded. The acceleration waveform for each dummy was recorded and used to calculate a dynamic response index (DRI) to assess the risk of spinal injury. Impacts with canopy fragments during ejections resulted in minor visible damages to the dummies and their equipment regardless of MDC firing. It was noted that MDC firing significantly attenuated the mean neck load (231 +/- 63 kg, unfired, to 108 +/- 20 kg, fired, p < 0.05). Noise levels with MDC firing averaged 162 dB. The mean DRI of dummy Gz for the small dummies was significantly greater than for large dummies (20.8 +/- 0.44 and 18.05 +/- 0.98, respectively, p < 0.05). We conclude that superficial injuries to ejecting pilots and their equipment is not a hazard with the canopy fragmentation system; however, spinal injury may occur at rates of 5 to 50% depending on the size of the pilot. PMID- 8424749 TI - Limited proteolysis of rat liver nucleolin by endogenous proteases: effects of polyamines and histones. AB - Nucleolin is a major nucleolar phosphoprotein and is presumably involved in rDNA transcription and ribosome biosynthesis. This protein is known to be very labile and to be cleaved by endogenous proteases into many small peptides. We found that, when rat liver nucleolar suspension (Nu-1) or nucleolin-rich extract (Nu-2) was incubated under conventional conditions, polyamines and histones interacted with the nucleolin to lead to its preferential degradation to 60 kDa phosphopeptide (p60). The peptide p60 was identified as a peptide containing the N-terminal half of the nucleolin molecule, as judged from peptide-map analysis. Whereas spermine binding to the purified nucleolin was decreased by KCl concentrations above 50 mM, histones (H1, H2B and H3) were able to bind to the nucleolin in the presence of up to 300 mM KCl. A distinct difference between H1 and other histones was found in that H1 could produce p60 from nucleolin in both Nu-1 and Nu-2, whereas H2B and H3 stimulated the degradation of nucleolin to p60 only when Nu-2 was used for the source of nucleolin. A possible relationship between p60 formation and rRNA synthesis is discussed, but its exact role remains to be studied. PMID- 8424750 TI - Agonist activation of transfected human M1 muscarinic acetylcholine receptors in CHO cells results in down-regulation of both the receptor and the alpha subunit of the G-protein Gq. AB - CHO cells stably transfected with cDNA encoding the human M1 muscarinic acetylcholine (HM1) receptor were treated with the cholinergic agonist carbachol at various concentrations for differing times. Levels of the HM1 receptor and of a range of G-proteins were subsequently measured. Carbachol treatment of the transfected cells caused a substantial down-regulation of cellular levels of the alpha subunit of Gq (Gq alpha), but did not significantly alter cellular levels of the alpha subunits of Gs or Gi2. A small decrease in levels of G-protein beta subunit was also produced. Parallel assessment of agonist-induced down-regulation of the HM1 receptor demonstrated that it was lost in concert with the G-protein. Similar concentrations of carbachol (5 microM) were required to produce half maximal stimulation of inositol phosphate generation and loss of each of the HM1 receptor and Gq alpha, and half-maximal losses of both receptor and Gq alpha were produced by 3 h of treatment with 1 mM-carbachol. By contrast, treatment of the non-transfected parental CHO cells, which do not express detectable levels of the receptor, with carbachol had no effect on cellular Gq alpha levels. Concurrent treatment of the HM1-expressing CHO cells with carbachol and cycloheximide indicated that suppression of protein synthesis de novo did not mimic the effect of carbachol, and hence even complete inhibition of transcription of the Gq alpha gene and/or translation of pre-existing Gq alpha mRNA could not account for the agonist-induced effect. We have previously noted that cellular levels of both Gs alpha [McKenzie and Milligan (1990) J. Biol. Chem. 265, 17084-17093] and the alpha subunits of the pertussis-toxin-sensitive G-proteins Gi1, Gi2 and Gi3 [Green, Johnson and Milligan (1990) J. Biol. Chem. 265, 5206-5210] can be regulated in certain cell systems by agonist activation of receptors expected to interact with these G-proteins. These results demonstrate that the same is true of Gq alpha and suggest that agonist-induced co-ordinate loss of receptors and associated G-proteins may be a more common feature than has been appreciated to date. PMID- 8424751 TI - Structural analysis of the human insulin-like growth factor-II P3 promoter. AB - The expression of insulin-like growth factor-II (IGF-II) has been observed previously in many human cancers. The human IGF-II P3 promoter has been shown by others to give rise to abundant 6.0 kb and 2.2 kb fetal transcripts which are expressed in a variety of both paediatric and adult tumours. In order to determine the mechanism by which the P3 promoter is controlled, the promoter was analysed in cell lines using chloramphenicol acetyltransferase (CAT) assay and DNAase I footprinting techniques. The data indicated that P3 is a complex promoter involving at least nine transcription factor binding sites. Furthermore, high levels of 5-methylcytosine detected in the P3 promoter of HeLa genomic DNA suggest that IGF-II gene expression may also be influenced by DNA methylation. PMID- 8424752 TI - Synthesis of antimony complexes of yeast mannan and mannan derivatives and their effect on Leishmania-infected macrophages. AB - Antimony(Sb)-yeast mannan complexes were synthesized as a strategy to introduce Sb into macrophages infected with Leishmania amastigotes. The complexes were taken up by endocytosis after specific recognition by alpha-D-mannosyl receptors on the macrophage membrane. About 90% of the intracellular parasites were destroyed by Sb-mannan in vitro, whereas the corresponding Sb concentration used as the pentavalent antimonial drug glucantime destroyed about 60% of the amastigotes. None of the Sb complexes prepared with mannan acid or basic derivatives was as effective as the simple Sb-mannan complex in clearing macrophage infection by Leishmania (L) amazonensis. The leishmanicidal effect of Sb-mannan was also demonstrated in vivo with infected hamsters. The alternative use of Sb-mannan complex in the treatment of human leishmaniasis is envisaged on the basis of parasite-killing efficiency and the use of a low antimony dose. PMID- 8424753 TI - Intramitochondrial control of the oxidation of hexadecanoate in skeletal muscle. A study of the acyl-CoA esters which accumulate during rat skeletal-muscle mitochondrial beta-oxidation of [U-14C]hexadecanoate and [U-14C]hexadecanoyl carnitine. AB - 1. We describe the acyl-CoA and acyl-carnitine esters which arise from the incubation of well-coupled State 3 rat skeletal-muscle mitochondrial fractions with [U-14C]hexadecanoate and [U-14C]hexadecanoyl-carnitine. 2. Acyl-CoA ester intermediates of chain length 16, 14, 12, 10 and 8 carbons were detected. 3. Although incubations were in steady state in respect of oxygen consumption, 14CO2 production and generation of acid-soluble radioactivity, quantitative analysis of acyl-CoA esters showed that steady state was not achieved in respect of all intermediates. 4. 3-Hydroxyacyl- and 2-enoyl-CoA and -carnitine esters were found under normoxic conditions. 5. Direct measurement of NAD+ and NADH shows that under identical incubation conditions our observations cannot be explained by gross perturbation of the [NAD+]/[NADH] ratio. 6. We hypothesize that there is a small pool of rapidly recycling NAD+ channelled between complex I of the respiratory chain and the newly described mitochondrial-inner-membrane-associated beta-oxidation trifunctional enzyme [Uchida, Izai, Orii and Hashimoto (1992) J. Biol. Chem. 267, 1034-1041]. PMID- 8424754 TI - The effect of treatment of the rat with bacterial endotoxin on gluconeogenesis and pyruvate metabolism in subsequently isolated hepatocytes. AB - The effect of treatment of rats with bacterial endotoxin on gluconeogenesis and the flux through pyruvate kinase, phosphoenolpyruvate carboxykinase (PEPCK), pyruvate carboxylase and pyruvate dehydrogenase (PDH) was measured in isolated hepatocytes, prepared from animals starved for 18 h, incubated in the presence of 1 mM pyruvate. The lipopolysaccharide reduced gluconeogenesis by 50% and lowered the flux through pyruvate kinase, PEPCK and pyruvate carboxylase by comparable amounts. There was no effect of endotoxaemia on PDH flux, indicating that the lowered rate of gluconeogenesis is not the result of a redistribution of pyruvate metabolism between oxidation and carboxylation. The results confirm that a stimulation of pyruvate kinase activity following treatment with lipopolysaccharide is not involved in the inhibition of gluconeogenesis, but that the effect resides at the level of phosphoenolpyruvate formation. The most favoured mechanism for the inhibition of glucose synthesis is via an inhibition of PEPCK and subsequent feedback inhibition of pyruvate carboxylase, although a secondary effect at the level of the mitochondria and pyruvate carboxylase cannot be excluded. PMID- 8424755 TI - Biology of the Rap proteins, members of the ras superfamily of GTP-binding proteins. PMID- 8424757 TI - Fluxionate Lewis acidity of the Zn2+ ion in carboxypeptidase A. AB - Competitive inhibition constants Ki for a series of phenol-ring-substituted derivatives of alpha-(2-hydroxyphenyl)benzenepropanoic acid have been ascertained by observing their influence on the catalytic hydrolysis of a peptide substrate by the zinc enzyme carboxypeptidase A. The pH-dependence of Ki shows that binding is maximal between two pKa values: one is that of the phenol group of the inhibitor, and the other uniformly has a value of 6, the pKa of a Zn(2+)-bound water molecule on the enzyme in the absence of substrate or inhibitor. This is the dependence expected if phenolate binds to the Zn2+ displacing its bound H2O/HO-. A log-log plot of the dissociation constants for the productive forms of inhibitor plus enzyme versus the acid dissociation constants of the phenolic residues in the inhibitors yields a straight line with a slope of +0.76. This number indicates that the active-site metal ion has special capacity for dispersing negative charge, such as builds up on the oxygen atom of a carboxamide group undergoing nucleophilic addition. PMID- 8424756 TI - Hyaluronate synthase: cloning and sequencing of the gene from Streptococcus sp. AB - The complete nucleotide sequence of hyaluronate synthase from Streptococcus sp. and its flanking regions is presented. The gene locus was designated has. Southern-blotting results suggested that the gene was conserved in hyaluronate producing streptococci. A putative translation-initiation codon was identified and the open reading frame consists of 1566 bp, specifying a protein of 56 kDa. Sequences resembling the promoter and ribosome-binding site of Gram-positive organisms are found upstream of the synthase. The predicted amino-acid sequence reveals the presence of a 35-residue signal peptide. The sequence has some similarity to bacterial peptide-binding proteins. PMID- 8424758 TI - Two cysteine substitutions in procollagen I: a glycine replacement near the N terminus of alpha 1(I) chain causes lethal osteogenesis imperfecta and a glycine replacement in the alpha 2(I) chain markedly destabilizes the triple helix. AB - Cultured skin fibroblasts were examined from two probands with type II (lethal) osteogenesis imperfecta. One proband had a single base mutation which converted the glycine codon at position alpha 1-244 in the alpha 1(I) chain of procollagen I into a cysteine codon whereas the other had a similar mutation that converted the glycine codon at position alpha 2-787 of the alpha 2(I) chain into a cysteine codon. Both mutations produced post-translational overmodification of procollagen I. The Cys alpha 1-244 mutation, however, had a minimal effect on the thermal stability or secretion of the protein whereas the Cys alpha 2-787 mutation markedly decreased the thermal stability and, apparently as a result, essentially none of the mutated protein was secreted. The results provide clear exceptions to two previous generalizations about the position-specificity of glycine substitutions in procollagen I. PMID- 8424759 TI - Arthrobacter D-xylose isomerase: partial proteolysis with thermolysin. AB - The pattern and kinetics of partial proteolysis of Arthrobacter D-xylose isomerase tetramer was studied in order to determine the flexibility of surface loops that may control its stability. It was completely resistant to trypsin, chymotrypsin and elastase at 37 degrees C, but thermolysin cleaved specifically and quantitatively at Thr-347-Leu-348 between helices 10 and 11 to remove 47 residues from the C-terminus of each 43.3 kDa subunit. At high temperatures, helices 9 and 10 were removed from each 38 kDa subunit to give a 36 kDa tetramer. The kinetics of nicking by thermolysin indicated that the Thr-347-Leu-348 loop is locked at low temperatures, but 'melts' at 25 degrees C and is fully flexible above 34 degrees C. The flexibility appears to be associated with binding of Ca2+ ions at the active site, since Co2+, Mg2+ and xylitol protect in proportion to their ability to displace Ca2+. The missing C-terminal helices make many intersubunit contacts that appear in the structure to stabilize the tetramer, but the properties of the purified nicked proteins are almost indistinguishable from the native enzyme. Both the 38 kDa tetramer and the 36 kDa tetramer are identically active and dissociate similarly in urea or SDS to fully active dimers, but the nicked dimers are slightly less stable to urea at 62 degrees C. In the Mg2+ form the thermostability of the 38 kDa tetramer is identical with that of the native enzyme, but the 36 kDa tetramer has a slightly lower 'melting point' (70 degrees C versus 80 degrees C), which may be due to unravelling from the end of helix 8. Since elimination of all the C-terminal helices and many intersubunit contacts has so little effect, one can conclude that the 'weak point' that controls the protein's thermostability lies within the N-terminal beta-barrel domain. PMID- 8424760 TI - Circular-dichroism analyses of membrane proteins: examination of environmental effects on bacteriorhodopsin spectra. AB - The secondary structure of bacteriorhodopsin is known from electron-diffraction studies, making bacteriorhodopsin a useful test system for analysing environmental influences on membrane proteins using c.d. spectroscopy. The conformational effects of detergent solubilization and incorporation into vesicles of various types were determined by comparison of the calculated secondary structures derived from c.d. spectra with the structure determined from diffraction studies. In addition, two modified forms of the native purple membrane, a shrunken form of the hexagonal lattice and an orthorhombic lattice form, were used to determine the effects of varying membrane fragment size and protein concentration within the membranes. The two different vesicle incorporation procedures yielded bacteriorhodopsin spectra which were nearly identical with each other and very close to the structure calculated from electron-diffraction studies. Solubilization of the native protein in the non ionic detergent n-octyl glucoside, without subsequent vesicle incorporation, resulted in a significantly altered protein conformation. Organizing the protein in different membrane lattices produced even more apparent deviations from the secondary structure determined by diffraction studies, as a consequence of optical effects caused by the high protein concentrations in the lattices. These studies show the importance of maintaining a 'native' environment, and the influence of particle geometry in interpreting c.d. studies of membrane proteins. PMID- 8424761 TI - Focal-adhesion components are enriched in ventral membranes isolated from transformed keratinocytes in culture. AB - The lack of procedures for isolating or reconstituting intact focal adhesions has hindered studies of how focal adhesions are organized and of how their assembly/disassembly is controlled. A method for isolating large quantities of the ventral portion of plasma membranes from transformed keratinocytes (A-431 cells) in culture is described. Plasma membranes are stabilized using Zn2+ and the ventral portion isolated attached to the culture substratum after the body of the cell has been sheared away. Compared with complete plasma membranes isolated from cells scraped from the dish, these ventral-membrane preparations are enriched 18.5-fold and 5.1-fold in the focal-adhesion components talin and vinculin respectively. While the epidermal-growth-factor receptor-kinase is less abundant in preparations of ventral membranes, a recently described tyrosine kinase which localizes to focal adhesions in mouse fibroblasts is enriched 19.9 fold. Extracellular matrix components, as well as their integrin receptors, are also enriched in these preparations of the ventral portion of plasma membranes compared with preparations of complete plasma membranes. PMID- 8424762 TI - Recombinant human iduronate-2-sulphatase: correction of mucopolysaccharidosis type II fibroblasts and characterization of the purified enzyme. AB - Mucopolysaccharidosis type II (MPS II, Hunter syndrome) is an X-chromosome-linked recessive lysosomal storage disorder that results from a deficiency of iduronate 2-sulphatase (12S). Patients with MPS II store and excrete large amounts of partially degraded heparan sulphate and dermatan sulphate. In order to evaluate enzyme-replacement therapy for MPS II we have expressed a chimaeric I2S cDNA in CHO (Chinese-hamster ovary)-K1 cells utilizing a plasmid vector that places the cDNA under the transcriptional control of the human polypeptide-chain-elongation factor-1 alpha gene promoter. A clonal cell line that accumulated recombinant I2S at greater than 10 mg/ml in conditioned medium was identified. Enzyme secreted from this cell line grown in the presence of NH4Cl was shown to be endocytosed into MPS II fibroblasts via the mannose 6-phosphate receptor and localized to the lysosomal compartment, resulting in correction of the storage phenotype of these cells. Milligram quantities of the recombinant I2S were purified, and the enzyme was shown to have a pH optimum and kinetic parameters similar to those for the mature form of I2S purified from human liver. The recombinant I2S had a molecular mass of approx. 90 kDa; this was reduced to 60 kDa by endoglycosidase treatment. PMID- 8424763 TI - The mouse collagen X gene: complete nucleotide sequence, exon structure and expression pattern. AB - Overlapping genomic clones covering the 7.2 kb mouse alpha 1(X) collagen gene, 0.86 kb of promoter and 1.25 kb of 3'-flanking sequences were isolated from two genomic libraries and characterized by nucleotide sequencing. Typical features of the gene include a unique three-exon structure, similar to that in the chick gene, with the entire triple-helical domain of 463 amino acids coded by a single large exon. The highest degree of amino acid and nucleotide sequence conservation was seen in the coding region for the collagenous and C-terminal non-collagenous domains between the mouse and known chick, bovine and human collagen type X sequences. More divergence between the sequences occurred in the N-terminal non collagenous domain. Similarity between the mammalian collagen X sequences extended into the 3'-untranslated sequence, particularly near the polyadenylation site. The promoter of the mouse collagen X gene was found to contain two TATAA boxes 159 bp apart; primer extension analyses of the transcription start site revealed that both were functional. The promoter has an unusual structure with a very low G + C content of 28% between positions -220 and -1 of the upstream transcription start site. Northern and in situ hybridization analyses confirmed that the expression of the alpha 1(X) collagen gene is restricted to hypertrophic chondrocytes in tissues undergoing endochondral calcification. The detailed sequence information of the gene is useful for studies on the promoter activity of the gene and for generation of transgenic mice. PMID- 8424764 TI - Purified yeast protein farnesyltransferase is structurally and functionally similar to its mammalian counterpart. AB - Protein farnesyltransferase (FTase) catalyses the addition of a farnesyl group to a cysteine within the so-called 'CAAX box' at the C-terminus of various proteins. In the present paper we report purification of Saccharomyces cerevisiae FTase to near-homogeneity. This was accomplished by constructing a yeast strain overproducing FTase approx. 100-fold. The purified enzyme was a heterodimer of approx. 90 kDa and consisted of 43 kDa and 34 kDa subunits. The 43 kDa subunit was shown to be the product of the DPR1 gene by using antibody raised against baculovirus-produced DPR1 polypeptide. The purified enzyme required Mg2+, showed a pH optimum of 7.8 and was most active at 50 degrees C. The Km values for farnesyl pyrophosphate and GST-CIIS (glutathione S-transferase fused to the C terminal 12 amino acids of yeast RAS2 protein), KmFpp and KmGST CIIS, were 8.1 and 5.1 microM respectively. The enzyme was capable of farnesylating GST-CIIL (the same as GST-CIIS, except that the C-terminal serine is changed to leucine), a substrate protein for the enzyme geranylgeranyltransferase, although with a higher apparent Km than for GST-CIIS. Like its mammalian counterpart, yeast FTase activity was inhibited by peptides containing the C-terminal CAAX sequence (that is, one where C = cysteine, A = aliphatic amino acid and X = any amino acid). These results provide direct evidence for the idea that the yeast and mammalian FTases are structurally and functionally very similar. PMID- 8424765 TI - A basolateral lactate/H+ co-transporter in Madin-Darby Canine Kidney (MDCK) cells. AB - Monolayers of Madin-Darby Canine Kidney (MDCK) cells grown on permeable filters generated lactate aerobically and accumulated it preferentially in the basolateral compartment, suggesting the presence of a lactate carrier. The mechanism of lactate transport across apical and basolateral membranes was examined by determining intracellular pH (pHi) microspectrofluorimetrically after addition of lactate to the extracellular solutions and by measuring uptake of [14C]lactate. Addition of 20 mM lactate to the apical compartment produced no change in pHi, whereas lactate added to the basolateral compartment rapidly and reversibly lowered pHi. Pyruvate produced similar results. Inhibitors of lactate/H+ co-transporters, alpha-cyano-4-hydroxycinnamate (CnCN) and quercetin, partially inhibited the fall in pHi produced by basolateral lactate. In contrast, the disulphonic stilbene. DIDS (4,4'-di-isothiocyanostilbene-2,2'-disulphonic acid) produced no inhibition at 0.5 mM. Kinetic analysis was performed by applying basolateral lactate at various concentrations and measuring the rate of entry (delta pHi/min) in the presence and absence of CnCN. Lactate flux was shown to occur by both non-ionic diffusion and a alpha-cyano-4-hydroxycinnamate sensitive component (carrier). The latter has a Km of approximately 7 mM for the lactate anion. Propionate, but not formate, lowered pHi to the same degree as did equimolar lactate, but the propionate effect was not inhibited by CnCN. Influx of [14C]lactate was substantially greater across the basolateral membrane than across the apical membrane and occurred in the absence of Na+. We conclude that MDCK cells grown on permeable filters generate lactate aerobically and transport it across the basolateral membrane by way of a lactate/H+ cotransporter. PMID- 8424766 TI - Synergy between Ca2+ and protein kinase C is the major factor in determining the level of secretion from human platelets. AB - The aim of this study was to establish further the role of protein kinase C in aggregation and secretion of 5-hydroxytryptamine (5-HT) from human platelets by using the selective inhibitor Ro 31-8220. Ro 31-8220 (3 microM) inhibited completely phosphorylation of pleckstrin, the major protein kinase C substrate, induced by thrombin, A23187 or phorbol dibutyrate (PDBu). Myosin light-chain phosphorylation induced by PDBu was also inhibited completely, but that induced by thrombin or A23187 was only inhibited partially. As myosin light chain is a substrate for both myosin light-chain kinase and protein kinase C, these results suggest that Ro 31-8220 is inhibiting only the protein kinase C-induced phosphorylation and that Ro 31-8220 has a greater selectivity to protein kinase C than does its structural analogue staurosporine. The stimulation of secretion of 5-HT by maximally effective concentrations of thrombin and A23187 was decreased significantly by 3 microM Ro 31-8220, but not inhibited completely. These results indicate a major role for protein kinase C in the stimulation of secretion by agonist- and ionophore-induced activation. On its own, a maximal concentration of PDBu induced a small degree of secretion (3.3 +/- 1.0%), but potentiated markedly the response to a submaximal concentration of A23187 (300 nM) to a level greater than seen with a maximal concentration of A23187. A similar set of results was also seen with aggregation, but not with shape change. We interpret these results to mean that the signalling event for secretion and aggregation is Ca2+, and this is potentiated markedly by protein kinase C. In the case of secretion, it appears that it is the synergy which is the major determining factor in influencing the extent. PMID- 8424767 TI - Physiological concentrations of inorganic phosphate affect MgATP-dependent Ca2+ storage and inositol trisphosphate-induced Ca2+ efflux in microsomal vesicles from non-hepatic cells. AB - 1. MgATP-dependent 45Ca2+ uptake by microsomes obtained from various non-hepatic tissues, namely rat brain, rat solid Morris hepatoma 3924A and human platelets, was measured in the presence of P(i) at low, cytosol-like, concentrations. 2. Increasing P(i) concentrations (0.5-3 mM) caused a progressive enlargement of the 45Ca(2+)-storage capacity of all the microsomal fractions. 3. As a result of P(i) stimulation of Ca2+ uptake, 45Ca2+ and [32P]P(i) were co-accumulated by the three microsomal fractions. 4. The time course for 45Ca2+ and [32P]P(i) accumulation in brain microsomes revealed a biphasic 45Ca2+ uptake: a rapid phase was followed by a second, slower, phase, which depended on the presence of P(i). During the P(i) dependent phase, the uptake of 45Ca2+ was paralleled by the uptake of [32P]Pi. 5. The passive efflux of Ca2+ was paralleled by the efflux of P(i) and vice versa. In fact, the inhibition of active Ca2+ uptake by excess EGTA, or lowering the P(i) concentration of the incubation system by dilution, caused the release of 45Ca2+ and [32P]P(i) from 45Ca2+ or [32P]P(i) pre-loaded brain microsomes. The Ca2+ ionophore A23187 also released 45Ca2+ and [32P]P(i). 6. Ca2+ efflux by A23187 was rapid (t 1/2 approx. 2 s) and independent of the extent of intravesicular Ca2+ loading, which indicates that Ca2+ and P(i) do not form intravesicular insoluble complexes. 7. The progressive increase in Ca2+ accumulation, depending on P(i) stimulation, resulted in a proportional increase in the amount of Ca2+ releasable by InsP3 in the three non-hepatic microsomal fractions and in digitonin-permeabilized platelets. 8. Concomitantly to Ca2+, microsomal P(i) was also released by InsP3. PMID- 8424768 TI - Effects of brefeldin A on sphingomyelin transport and lipid synthesis in BHK21 cells. AB - 1. Addition of brefeldin A (BFA) to BHK cells incubated for 4 h with [3H]acetate led to a 3-4-fold increase in incorporation of label into sphingomyelin, monoglucosylceramide and cholesterol ester compared with untreated controls. There was a similar increase in incorporation of [3H]choline into sphingomyelin. The level of cholesterol ester increased 3-fold when BFA was added to cells labelled to equilibrium with [3H]acetate, but no statistically significant changes in the levels of other lipids were seen. 2. BFA appeared to act by diverting incorporation of acetate into sphingolipids and cholesterol ester at the expense of phosphatidylcholine (decreased by up to 15%), cholesterol (decreased by 30-40%) and triacylglycerol (decreased by 35-50%). 3. Forskolin (100 microM) prevented the changes in labelling induced by 0.25 micrograms of BFA/ml, but in the presence of 1 micrograms of BFA/ml it had no effect on sphingomyelin and triacylglycerol labelling and only partly blocked the effects of BFA on labelling of cholesterol and cholesterol ester. 4. None of the labelled sphingomyelin was degraded in BFA-treated cells which were subsequently exposed to an extracellular sphingomyelinase, showing that all the newly synthesized sphingomyelin remained inside the cells. Determinations of phospholipid phosphorus in unlabelled cells confirmed that, in the presence of BFA, no newly synthesized sphingomyelin was able to reach the cell surface, supporting the idea that sphingomyelin normally depends on vesicular transport for its passage to the plasma membrane. 5. The results are consistent with the hypothesis that cholesterol synthesis and esterification processes in BHK cells are sensitive to the plasma-membrane deficit of sphingomyelin caused by BFA. PMID- 8424769 TI - Hepatic cholesterol synthesis and the secretion of newly synthesized cholesterol in bile. AB - To determine the effect of increased hepatic cholesterol synthesis on the secretion of newly synthesized cholesterol in bile, rats were fed with cholestyramine, a bile-acid-binding resin that increases the number of hepatocytes that synthesize cholesterol. Cholesterol synthesis was measured 15 min after [3H]water injection to avoid appreciable exchange between the liver and serum of newly synthesized cholesterol that accumulates in the serum in studies of several hours duration. At 15 min after [3H]water injection, the specific radioactivity of cholesterol in the liver and hepatic microsomes was greatly increased in resin-fed animals compared with controls. However, with resin, the specific radioactivity of newly synthesized cholesterol that was secreted in bile was the same as for controls. At 15 min after [3H]water injection the specific radioactivity of serum cholesterol was minimally increased and not different in resin and control groups. In contrast, in studies that were longer than 60 min, newly synthesized cholesterol in serum was appreciably increased in resin-fed animals, and newly synthesized cholesterol in bile was also greatly increased compared with controls. Thus, when appreciable cholesterol exchange is avoided, an increase in hepatic cholesterol synthesis and the number of hepatocytes that synthesized cholesterol does not result in an increase in newly synthesized cholesterol in bile. Our results suggest that newly synthesized cholesterol is secreted in bile from a fixed subpopulation of hepatocytes. From a comparison of the specific radioactivity of newly synthesized cholesterol in whole liver and bile, it can be estimated that this subpopulation of hepatocytes represents about 20% of the total hepatocyte mass. PMID- 8424770 TI - Leupeptin-binding site(s) in the mammalian multicatalytic proteinase complex. AB - The multicatalytic proteinase (MCP) complex is a major nonlysosomal proteinase which plays an important role in non-lysosomal pathways of protein degradation and which has recently been implicated in antigen processing. The mammalian MCP complex is composed of more than 20 different types of polypeptide, but it is not yet clear which of these components are responsible for its proteolytic activities. The complex has at least three distinct types of proteolytic activity. One of these, the so-called 'trypsin-like' activity, which involves cleavage on the carboxy side of basic amino acid residues, can be selectively and completely inhibited by peptidyl arginine aldehydes (such as leupeptin and antipain), and is also the most sensitive to inhibition by thiol-reactive reagents. In the present study N-[ethyl-1-14C]ethylmaleimide has been used to specifically label thiol groups protected by leupeptin binding. The results suggest that one or two polypeptide components within the complex can be protected against modification by N-ethylmaleimide. These components may be responsible for the 'trypsin-like' activity of the complex or may be adjacent to the catalytic component(s) and play an important role in substrate binding. PMID- 8424771 TI - Monitoring of changes in hepatic fatty acid and glycerolipid metabolism during the starved-to-fed transition in vivo. Studies on awake, unrestrained rats. AB - 1. The technique of selective labelling of hepatic fatty acids in vivo [Moir and Zammit (1992) Biochem. J. 283, 145-149] has been used to monitor non-invasively the metabolism of fatty acids in the livers of awake unrestrained rats during the starved-to-refed transition. Values for the incorporation of labelled fatty acid into liver and plasma glycerolipids and into exhaled carbon dioxide after injection of labelled lipoprotein and Triton WR 1339 into rats with chronically cannulated jugular veins were obtained for successive 1 h periods from the start of refeeding of 24 h-starved rats. 2. Starvation for 24 h resulted in marked and reciprocal changes in the incorporation of label into glycerolipids and exhaled 14CO2, such that a 4-fold higher value was obtained for the oxidation/esterification ratio in livers of starved rats compared with fed animals. 3. Refeeding of starved rats did not return this ratio to the value observed for fed animals for at least 7 h; during the first 3 h of refeeding the ratio was at least as high as that for starved rats. Between 4 h and 6 h of refeeding the ratio was still approx. 70% of that in starved animals, and 2.5 fold higher than in fed rats. 4. These data support the hypothesis that the capacity of the liver to oxidize fatty acids is maintained at a high level during the initial stages of refeeding [Grantham and Zammit (1986) Biochem. J. 239, 485 488] and that control of the flux of hepatic fatty acids into the oxidative pathway is largely lost from the reaction catalysed by mitochondrial overt carnitine palmitoyltransferase (CPT I) during this phase of recovery from the starved state. 5. Refeeding also resulted in a rapid (< 1 h) increase in hepatic malonyl-CoA concentrations to values intermediate between those in livers of fed and starved animals. The sensitivity of CPT I to malonyl-CoA inhibition in isolated liver mitochondria was only partially reversed even after 5 h of refeeding. 6. Refeeding resulted in an acute 35% inhibition of the fraction of synthesized triacylglycerol that was secreted into the plasma; the maximal effect occurred 2-3 h after the start of refeeding. The inhibition of the fractional secretion rate was fully reversed after 5 h of refeeding. 7. The amount of 14C label that was incorporated into phospholipids as a fraction of total glycerolipid synthesis was doubled within 2 h of the start of refeeding.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8424772 TI - Altered protein secretion and extracellular matrix deposition is associated with the proliferative phenotype induced by allylamine in aortic smooth muscle cells. AB - Repeated cycles of allylamine-induced aortic injury in vivo modulate the proliferative potential of smooth muscle cells (SMCs) during serial propagation in vitro. This modulation may be partly mediated by disturbances in polyphosphoinositide metabolism which afford allylamine-treated cells a growth advantage over control cells [Cox, Murphy and Ramos (1990) Exp. Mol. Pathol. 53, 52-63]. The present studies were conducted to further evaluate the mechanisms which mediate the enhanced proliferative potential of allylamine cells. Cellular growth and/or [3H]thymidine incorporation into DNA were evaluated in control and allylamine cells seeded on plastic culture dishes or glass coverslips in the presence of 0.1, 1 or 10% fetal bovine serum (FBS). On either substrate, incubation in 0.1% FBS for 48 h inhibited DNA synthesis in cultures of both cell types, but the inhibitory response was more pronounced in allylamine cells. Subsequent challenge with 10% FBS increased thymidine incorporation to a greater extent in allylamine cells. Interestingly, enhanced DNA synthesis of allylamine cells was associated with increased cell numbers only when seeded on a glass surface. The enhanced growth rate on glass was not due to increased plating efficiency since comparable attachment rates were observed for both cell types. Reseeding of control cells on glass substrates pre-coated by allylamine cells afforded control cells a growth advantage comparable with that observed for allylamine cultures. Conditioned media from growth-arrested, as well as cycling cultures, of allylamine cells stimulated DNA synthesis in cultures of either cell type to a greater extent than conditioned media from control counterparts. In addition, the responsiveness of allylamine cells to secreted products was enhanced relative to that of control cells. Metabolic labelling studies revealed that the synthesis and/or secretion of 52, 46, 33 and 28 kDa proteins was enhanced in allylamine cultures relative to controls, and that the expression of two proteins of 30 and 31 kDa only occurred in allylamine cultures. We conclude that the enhanced growth response of allylamine cells is associated with both altered protein secretion and differential extracellular matrix deposition. PMID- 8424773 TI - Characterization of the functional domain of tissue inhibitor of metalloproteinases-2 (TIMP-2). AB - Analysis of the functional domain of tissue inhibitor of metallo-proteinases-2 (TIMP-2) was performed using limited proteolytic degradation with trypsin. This treatment generated a 13.5 kDa fragment which was purified and shown to consist of an uncleaved N-terminal region extending from residue 1 to residue 132. The fragment retains the ability to inhibit activated interstitial collagenase and to block the autocatalytic activation of procollagenase. PMID- 8424774 TI - Inhibition of protein synthesis and early protein processing by thapsigargin in cultured cells. AB - Thapsigargin, a tumour-promoting sesquiterpene lactone, selectively inhibits the Ca(2+)-ATPase responsible for Ca2+ accumulation by the endoplasmic reticulum (ER). Mobilization of ER-sequestered Ca2+ to the cytosol and to the extracellular fluid subsequently ensues, with concomitant alteration of cellular functions. Thapsigargin was found to serve as a rapid, potent and efficacious inhibitor of amino acid incorporation in cultured mammalian cells. At concentrations mobilizing cell-associated Ca2+ to the extracellular fluid, thapsigargin provoked extensive inhibition of protein synthesis within 10 min. The inhibition in GH3 pituitary cells involved the synthesis of almost all polypeptides, was not associated with increased cytosolic free Ca2+ concentration ([Ca2+]i), and was not reversed at high extracellular Ca2+. The transient rise in [Ca2+]i triggered by ionomycin was diminished by thapsigargin. Polysomes failed to accumulate in the presence of the drug, indicative of impaired translational initiation. With longer (1-3 h) exposures to thapsigargin, recovery of translational activity was observed accompanied by increased synthesis of the ER protein glucose-regulated stress protein 78 or immunoglobulin heavy-chain binding protein ('GRP78/BiP') and its mRNA. Such inductions were comparable with those observed previously with Ca2+ ionophores which mobilize the cation from all intracellular sequestered sites. Actin mRNA concentrations declined significantly during such treatments. In HepG2 cells processing and secretion of the glycoprotein alpha 1-antitrypsin were rapidly suppressed by thapsigargin. Ca2+ sequestered specifically by the ER is concluded to be essential for optimal protein synthesis and processing. These rapid effects of thapsigargin on mRNA translation, protein processing and gene expression should be considered when evaluating potential mechanisms by which this tumour promoter influences cellular events. PMID- 8424775 TI - Expression and lipoylation in Escherichia coli of the inner lipoyl domain of the E2 component of the human pyruvate dehydrogenase complex. AB - The dihydrolipoamide acetyltransferase subunit (E2p) of mammalian pyruvate dehydrogenase complex has two highly conserved lipoyl domains each modified with a lipoyl cofactor bound in amide linkage to a specific lysine residue. A sub-gene encoding the inner lipoyl domain of human E2p has been over-expressed in Escherichia coli. Two forms of the domain have been purified, corresponding to lipoylated and non-lipoylated species. The apo-domain can be lipoylated in vitro with partially purified E. coli lipoate protein ligase, and the lipoylated domain can be reductively acetylated by human E1p (pyruvate dehydrogenase). Availability of the two forms will now allow detailed biochemical and structural studies of the human lipoyl domains. PMID- 8424776 TI - Purification of the pro-phenol oxidase enzyme from haemocytes of the cockroach Blaberus discoidalis. AB - Pro-phenol oxidase was purified from the haemocytes of the cockroach Blaberus discoidalis by Blue Sepharose chromatography, hydrophobic-interaction chromatography on a Phenyl-Superose column and, finally, gel filtration on a Superose 6 column. Results suggest that the molecule exists as a polymer of identical 76 kDa monomeric units. The enzyme is a glycoprotein with pI of 5.2 and can be converted by trypsin into phenol oxidase. PMID- 8424777 TI - Affinity labelling of the Ca(2+)-activated neutral proteinase (calpain) in intact human platelets. AB - Two irreversible calpain inhibitors, benzyloxycarbonyl (Cbz)-Leu-Leu-Tyr-Ch2F and Cbz-Leu-Leu-Tyr-CHN2, were shown earlier [Anagli, Hagmann and Shaw (1991) Biochem. J. 274, 497-502] to penetrate intact platelets and to inactivate calpain. This permitted an evaluation of certain functions attributed to this proteinase. For example, in platelets pretreated with these inhibitors, talin and actin-binding protein were protected from subsequent degradation when the Ca2+ level was raised. On the other hand, additional properties of stimulated platelets attributed to calpain remained unaffected by this treatment, and such hypotheses may be dismissed. Radioiodinated inhibitors permitted confirmation of the labelling of calpain by the procedures used. Although Cbz-Leu-Leu-Tyr-CHN2 is more effective in vitro than the corresponding fluoromethyl ketone, we now show that the latter penetrates more readily. These two inhibitors, and two additional ones, t-butyloxycarbonyl-Val-Lys(Cbz)-Leu-Tyr- CHN2 and Cbz-Leu-Tyr-CH2F, have been radioiodinated to permit a comparison of their intracellular labelling patterns in activated platelets. Calpain is the major target of all four inhibitors. Although they are closely related peptide structures, variations with respect to the labelling of additional proteins were observed. These were minor in the case of the peptidyl diazomethyl ketones, but were major in the case of the fluoromethyl ketones. However, in contrast to calpain, this labelling was neither time-dependent nor Ca(2+)-dependent. Radiolabelling and cellular fractionation studies were used to localize active calpain during platelet activation. Calpain appears to be activated in the cytosol and translocated to the membrane or cytoskeletal sites. PMID- 8424778 TI - Carrageenans inhibit growth-factor binding. AB - Carrageenans, a family of polysulphated carbohydrates, inhibited binding of basic fibroblast growth factor (bFGF), transforming growth factor beta 1 (TGF beta 1) and platelet-derived growth factor (PDGF). iota-Carrageenan was the most potent bFGF antagonist (IC50 = 0.4 +/- 0.1 microgram/ml), kappa-carrageenan was the most potent PDGF antagonist (IC50 = 1.7 +/- 1.3 micrograms/ml) and lambda-carrageenan was the most potent TGF beta 1 antagonist (IC50 = 19 +/- 2 micrograms/ml). None of the carrageenans, at concentrations up to 200 micrograms/ml, inhibited binding of insulin-like growth factor 1 or transforming growth factor alpha. Carrageenans are selective growth-factor antagonists and have potential for the treatment of disorders associated with the over-production of certain growth factors. PMID- 8424780 TI - Prenyl group identification of rap2 proteins: a ras superfamily member other than ras that is farnesylated. AB - Rap proteins comprise a subset of the large family of ras-related proteins. They contain the C-terminal tetrapeptide sequence motif Cys-Ali-Ali-Xaa (Ali is an aliphatic amino acid and X is any amino acid), which has been found to be the site of membrane attachment via isoprenylation for ras, nuclear lamins and the gamma subunits of the heterotrimeric G-proteins. To investigate the isoprenylation of rap2a and rap2b, human cDNAs coding for these proteins were expressed in COS cells incubated in the presence of [3H]mevalonolactone. Both proteins incorporated a product of [3H]mevalonolactone, as judged by Western blot analysis. To identify the specific isoprenoid attached to each protein, the cDNAs were transcribed in vitro and the rap2 specific RNA was translated in a rabbit reticulocyte lysate system in the presence of [3H]mevalonolactone. The translation products were treated with methyl iodide and the released isoprenoid groups were analysed by h.p.l.c. Rap2b, which terminates in Cys-Val-Ile-Leu, is geranylgeranylated as predicted while rap2a, which terminates in Cys-Asn-Ile-Gln, incorporated farnesyl. A mutant construct generated by site-directed mutagenesis of rap2a cDNA yielding a protein terminating in leucine instead of glutamine incorporated geranylgeranyl, lending further support to the notion that isoprenoid specificity is governed by the terminal amino acid. In addition, when the CAAX motif cysteine at position 180 of rap2a was replaced by a serine residue no isoprenoid incorporation was observed. Thus rap2a and rap2b, despite showing 90% sequence identity, incorporate different isoprenoid groups. Thus glutamine is a signal for farnesylation, and rap2a is the first non-ras member of the ras superfamily that is farnesylated. PMID- 8424779 TI - Bovine kidney beta-mannosidase: purification and characterization. AB - Lysosomal beta-mannosidase was purified 160,000-fold in 24% yield from bovine kidney by a four-step purification procedure, which included concanavalin A Sepharose, immunoaffinity, TSK-butyl and h.p.l.c. cation-exchange chromatography. When analysed by SDS/PAGE and detected by Coomassie Blue or silver staining, the purified enzyme preparation consists of two prominent peptides (100 and 110 kDa) and a third minor peptide (84 kDa). These three peptides are immunologically related and are consistently associated with beta-mannosidase activity in all chromatographic steps. Removal of N-linked carbohydrate from the 84, 100 and 110 kDa peptides decreases their molecular sizes to 75, 86 and 91 kDa respectively. Bovine kidneys lacking beta-mannosidase, activity, acquired from calves affected with beta-mannosidosis, do not contain detectable quantities of the three beta mannosidase peptides, as judged by monoclonal- and polyclonal-antibody reactivity. PMID- 8424781 TI - X-ray-crystallographic studies of complexes of pepstatin A and a statine containing human renin inhibitor with endothiapepsin. AB - H-189, a synthetic human renin inhibitor, and pepstatin A, a naturally occurring inhibitor of aspartic proteinases, have been co-crystallized with the fungal aspartic proteinase endothiapepsin (EC 3.4.23.6). H-189 [Pro-His-Pro-Phe-His-Sta (statyl)-Val-Ile-His-Lys] is an analogue of human angiotensinogen. Pepstatin A [Iva(isovaleryl)-Val-Val-Sta-Ala-Sta] is a blocked pentapeptide which inhibits many aspartic proteinases. The structures of the complexes have been determined by X-ray diffraction and refined to crystallographic R-factors of 0.15 and 0.16 at resolutions of 0.18 nm (1.8 A) and 0.2 nm (2.0 A) respectively. H-189 is in an extended conformation, in which the statine residue is a dipeptide analogue of P1 and P'1 as indicated by the conformation and network of contacts and hydrogen bonds. Pepstatin A has an extended conformation to the P'2 alanine residue, but the leucyl side chain of the terminal statine residue binds back into the S'1 subsite, and an inverse gamma-turn occurs between P'1 and P'3. The hydroxy moiety of the statine at P1 in both complexes displaces the solvent molecule that hydrogen-bonds with the catalytic aspartate residues (32 and 215) in the native enzyme. Solvent molecules originally present in the native structure at the active site are displaced on inhibitor binding (12 when pepstatin A binds; 16 when H-189 binds). PMID- 8424782 TI - Changes in free cytoplasmic magnesium following activation of human lymphocytes. AB - Activation of lymphocytes with 10 microM ionomycin leads to a rapid increase in the concentration of free cytoplasmic calcium ([Ca2+]i) and, at a slower rate, also to an increase in the cytoplasmic free magnesium concentration ([Mg2+]i). The ionomycin-induced Mg(2+)-mobilization response is dependent on the influx of extracellular Ca2+. After receptor-mediated lymphocyte activation, induced by mitogens or anti-receptor antibodies, a Mg(2+)-mobilization response does occur in a small fraction of the cells. Simultaneous measurement of [Ca2+]i and [Mg2+]i in individual cells showed that the receptor-triggered Mg(2+)-mobilization response is restricted to cells that have a high [Ca2+]i. It can therefore be concluded that a high [Ca2+]i induces the release into the cytoplasm of Mg2+ from intracellular stores. PMID- 8424783 TI - Blockade of mevalonate production by lovastatin attenuates bombesin and vasopressin potentiation of nutrient-induced insulin secretion in HIT-T15 cells. Probable involvement of small GTP-binding proteins. AB - Small G-proteins (SMGs) require isoprenylation for their association with membranes. We have examined protein isoprenylation, subcellular distribution of SMGs, cytosolic Ca2+ changes and insulin secretion in HIT-T15 cells after treatment with lovastatin, which inhibits the production of isoprenoids by blocking mevalonate production by 3-hydroxy-3-methylglutaryl-CoA reductase. Numerous proteins in the 20-70 kDa range were found to be isoprenylated. Most of these proteins co-migrated with SMGs (21-27 kDa). Lovastatin treatment (25 microM, 24 h) decreased protein isoprenylation and affected the distribution of several SMGs, causing a large accumulation in the cytosol and a detectable decrease in membranes. Lovastatin selectively attenuated the potentiating action of bombesin and vasopressin, which activate phospholipase C in these cells, on insulin secretion stimulated by nutrients (glucose + leucine + glutamine). This lovastatin effect was overcome by mevalonate. Insulin secretion stimulated by nutrients alone or insulin release in the presence of the potentiating agents forskolin or phorbol myristate acetate remained unaffected. As the modulation of insulin secretion by isoprenaline and somatostatin were not altered by lovastatin, the drug does not non-selectively affect the binding of ligands to their receptors. Lovastatin did not interfere with the activation of phospholipase C by bombesin and vasopressin, since the rise in cytosolic Ca2+ induced by these agents was not changed. Limonene, proposed to block specifically prenyl-protein transferases of SMGs, did not alter protein isoprenylation patterns, but inhibited the stimulated insulin secretion. In conclusion, lovastatin selectively attenuated the potentiation of nutrient-induced insulin secretion by bombesin and vasopressin without affecting their activation of phospholipase C. The concomitant changes in SMG isoprenylation and their subcellular distribution after lovastatin treatment suggest that SMGs could play an important role in the bombesin and vasopressin action on insulin secretion. PMID- 8424784 TI - G-proteins are not directly involved in the CD3-antigen-mediated production of inositol phosphates in HPB-ALL T-leukaemia cells expressing phospholipase C isoforms gamma 1 and beta 3. AB - The possible involvement of G-proteins in T cell antigen-receptor complex (TCR) mediated inositol phosphate production was investigated in HPB-ALL T-cells, which were found to express the phospholipase C gamma 1 and beta 3 isoforms. Cross linking the CD3 antigen on streptolysin-O-permeabilized cells stimulated a dose dependent increase in inositol phosphate production, as did addition of guanosine 5'-[gamma-thio]triphosphate (GTP[S]) or vanadate, a phosphotyrosine phosphatase inhibitor. It was possible, therefore, that the CD3-antigen-mediated production of inositol phosphates was either via a G-protein-dependent mechanism or by stimulation of protein tyrosine phosphorylation. The CD3-induced inositol phosphate production was potentiated by addition of vanadate, but not by addition of GTP[S]. Guanosine 5'-[beta-thio]diphosphate (GDP[S]) inhibited the rise in inositol phosphates induced by GTP[S], vanadate or cross-linking the CD3 antigen. The increase in protein tyrosine phosphorylation stimulated by vanadate or the OKT3 monoclonal antibody was not observed in the presence of GDP[S], showing that in permeabilized HPB-ALL cells, GDP[S] inhibits the actions of tyrosine kinases as well as G-protein function. Addition of either ADP[S] or phenylarsine oxide inhibited CD3- and vanadate-mediated increases in both tyrosine phosphorylation and inositol phosphate production, but did not inhibit GTP[S]-stimulated inositol phosphate production. On the other hand, pretreatment of cells with phorbol 12,13 dibutyrate inhibited subsequent GTP[S]-stimulated inositol phosphate production but did not inhibit significantly inositol phosphate production stimulated by either OKT3 F(ab')2 fragments or vanadate. Our results are consistent with the CD3 antigen stimulating inositol phosphate production by increasing the level of protein tyrosine phosphorylation, but not by activating a G-protein. PMID- 8424785 TI - The molybdenum and vanadium nitrogenases of Azotobacter chroococcum: effect of elevated temperature on N2 reduction. AB - During the reduction of N2 by V-nitrogenase at 30 degrees C, some hydrazine (N2H4) is formed as a product in addition to NH3 [Dilworth and Eady (1991) Biochem. J. 277, 465-468]. We show here the following. (1) That over the temperature range 30-45 degrees C the apparent Km for the reduction of N2 to yield these products is the same, but increases from 30 to 58 kPa of N2. On increasing the temperature from 45 degrees C to 50 degrees C, little change occurred in the rate of reduction of protons to H2; the rate of N2H4 production increased, but the rate of NH3 formation decreased 7-fold. (2) Temperature-shift experiments from 42 to 50 degrees C or from 50 to 42 degrees C showed that this selective loss of the ability to reduce N2 to NH3 was reversible. The effects we observe are consistent with the existence of different conformers of the VFe protein at the two temperatures, that predominating at 50 degrees C being largely unable to reduce N2 to ammonia. (3) Measurement of the ratio between H2 evolution and N2 reduced to NH3 at N2 pressures up to 339 kPa for both Mo- and V nitrogenases gave limiting H2/N2 values of 1.13 +/- 0.13 for Mo-nitrogenase and 3.50 +/- 0.03 for V-nitrogenase. Since for Mo-nitrogenase our measured value for the ratio at 339 kPa is the same as that derived by Simpson and Burris [(1984) Science 224, 1095-1097] at 5650 kPa, there appears to be little or no divergence from the predictions based on the apparent Km for N2. These data then suggest that there may be a fundamentally different mechanism for N2 binding to V nitrogenase compared with Mo-nitrogenase. (4) We did not detect any N2H4 as a product of N2 reduction by Mo-nitrogenase over the temperature range investigated; however, at 50 degrees C this system reduced acetylene (C2H2) to yield some ethane (C2H6), in addition to ethylene (C2H4), a reaction normally associated with Mo-independent nitrogenases. PMID- 8424786 TI - Purification and characterization of the major glutathione transferase from adult toad (Bufo bufo) liver. AB - Five forms of glutathione transferase (GST) were resolved from the cytosol of adult common toad (Bufo bufo) liver by GSH-affinity chromatography followed by isoelectric focusing. The major enzyme (GST-7.64; 55% of total activity bound to the column) has a pI value of 7.64, is composed of two subunits each with a molecular mass of 23 kDa, and has the N-terminal amino acid residue blocked. GST 7.64 has also been characterized with respect to amino acid composition, substrate specificity, inhibition characteristics, c.d. spectra and immunological reactivity. The N-terminal sequence of some peptides obtained after tryptic digestion has also been determined. All together the results obtained suggest that the major toad liver GST is distinct from any known GST, including microbial, plant and mammalian GSTs. PMID- 8424787 TI - Contractile activity restores insulin responsiveness in skeletal muscle of obese Zucker rats. AB - Both insulin and contraction stimulate glucose transport in skeletal muscle. Insulin-stimulated glucose transport is decreased in obese humans and rats. The aims of this study were (1) to determine if contraction-stimulated glucose transport was also compromised in skeletal muscle of genetically obese insulin resistant Zucker rats, and (2) to determine whether the additive effects of insulin and contraction previously observed in muscle from lean subjects were evident in muscle from the obese animals. To measure glucose transport, hindlimbs from lean and obese Zucker rats were perfused under basal, insulin-stimulated (0.1 microM), contraction-stimulated (electrical stimulation of the sciatic nerve) and combined insulin-(+)contraction-stimulated conditions. One hindlimb was stimulated to contract while the contralateral leg served as an unstimulated control. 2-Deoxyglucose transport rates were measured in the white gastrocnemius, red gastrocnemius and extensor digitorum longus muscles. As expected, the insulin stimulated glucose transport rate in each of the three muscles was significantly slower (P < 0.05) in obese rats when compared with lean animals. When expressed as fold stimulation over basal, there was no significant difference in contraction-induced muscle glucose transport rates between lean and obese animals. Insulin-(+)contraction-stimulation was additive in skeletal muscle of lean animals, but synergistic in skeletal muscle of obese animals. Prior contraction increased insulin responsiveness of glucose transport 2-5-fold in the obese rats, but had no effect on insulin responsiveness in the lean controls. This contraction-induced improvement in insulin responsiveness could be of clinical importance to obese subjects as a way to improve insulin-stimulated glucose uptake in resistant skeletal muscle. PMID- 8424788 TI - Substrate selectivities differ for hepatic mitochondrial and peroxisomal beta oxidation in an Antarctic fish, Notothenia gibberifrons. AB - Hepatic mitochondrial and peroxisomal beta-oxidation were examined in an Antarctic marine teleost, Notothenia gibberifrons. Enzymic profiles and rates of beta-oxidation by intact organelles were determined by using a range of fatty acyl-CoA substrates to evaluate substrate preferences. Partitioning of beta oxidation between organelles was estimated. Substrate selectivities are broader for peroxisomal beta-oxidation than for mitochondrial beta-oxidation. Mitochondria show marked preference for the oxidation of a monounsaturated substrate, palmitoleoyl-CoA (C16:1), and two polyunsaturates, eicosapentaenoyl CoA (C20:5) and docosahexaenoyl-CoA (C22:6). Carnitine palmitoyltransferase activities with palmitoleoyl-CoA (C16:1) are 2.4-fold higher than activities with palmitoyl-CoA (C16:0). Most polyunsaturated acyl-CoA esters measured appear to inhibit by over 40% the oxidation of palmitoyl-CoA by peroxisomes. Our findings suggest that the polyunsaturates, eicosapentaenoic acid (C20:5) and docosahexaenoic acid (C22:6), found in high concentrations in Antarctic fishes [Lund and Sidell (1992) Mar. Biol. 112, 377-382], are utilized as fuels to support aerobic energy metabolism. Metabolic capacities of rate-limiting enzymes and beta-oxidation rates by intact organelles indicate that up to 30% of hepatic beta-oxidation in N. gibberifrons can be initiated by the peroxisomal pathway. PMID- 8424789 TI - Evidence for novel binding sites on the platelet glycoprotein IIb and IIIa subunits and immobilized fibrinogen. AB - The present study was designed to examine the interaction of the purified platelet glycoprotein IIb-IIIa complex (GP IIb-IIIa or integrin alpha IIb beta 3) and the individual subunits of the complex with immobilized fibrinogen. Although 125I-GP IIb-IIIa binding to fibrinogen immobilized on Sepharose was specific, this interaction exhibited properties distinct from those of reversible fibrinogen binding to platelets: 125I-GP IIb-IIIa binding appeared irreversible, but non-covalent, Ca(2+)-independent, and was inhibited only weakly, or not at all, by the anti-(GP IIb-IIIa) monoclonal antibodies 10E5 and 7E3 and synthetic peptides from known platelet-binding domains of fibrinogen. Reversibly dissociated GP IIb or GP IIIa subunits inhibited 125I-GP IIb-IIIa binding to immobilized fibrinogen and bound directly to the fibrinogen. However, these subunits did not bind to peptides derived from known platelet-binding domains within the fibrinogen alpha- and gamma-chains, although the GP IIb-IIIa complex did. These results show that the complexed form of full-length GP IIb and GP IIIa is required for binding to these synthetic peptides, but not necessarily for binding to immobilized fibrinogen. Thus GP IIb-IIIa can bind to immobilized fibrinogen by a distinct mechanism that appears to involve novel binding sites on each subunit of the GP IIb-IIIa complex and on fibrinogen. PMID- 8424790 TI - Purification and properties of three (1-->3)-beta-D-glucanase isoenzymes from young leaves of barley (Hordeum vulgare). AB - Three (1-->3)-beta-D-glucan glucanohydrolase (EC 3.2.1.39) isoenzymes GI, GII and GIII were purified from young leaves of barley (Hordeum vulgare) using (NH4)2SO4 fractional precipitation, ion-exchange chromatography, chromatofocusing and gel filtration chromatography. The three (1-->3)-beta-D-glucanases are monomeric proteins of apparent M(r)32,000 with pI values in the range 8.8-10.3. N-terminal amino-acid-sequence analyses confirmed that the three isoenzymes represent the products of separate genes. Isoenzymes GI and GII are less stable at elevated temperatures and are active over a narrower pH range than is isoenzyme GIII, which is a glycoprotein containing 20-30 mol of hexose equivalents/mol of enzyme. The preferred substrate for the enzymes is laminarin from the brown alga Laminaria digitata, an essentially linear (1-->3)-beta-D-glucan with a low degree of glucosyl substitution at 0-6 and a degree of polymerization of approx. 25. The three enzymes are classified as endohydrolases, because they yield (1-->3)-beta-D oligoglucosides with degrees of polymerization of 3-8 in the initial stages of hydrolysis of laminarin. Kinetic analyses indicate apparent Km values in the range 172-208 microM, kcat. constants of 36-155 s-1 and pH optima of 4.8. Substrate specificity studies show that the three isoenzymes hydrolyse substituted (1-->3)-beta-D-glucans with degrees of polymerization of 25-31 and various high-M(r), substituted and side-branched fungal (1-->3;1-->6)-beta-D glucans. However, the isoenzymes differ in their rates of hydrolysis of a (1- >3;1-->6)-beta-D-glucan from baker's yeast and their specific activities against laminarin vary significantly. The enzymes do not hydrolyse (1-->3;1-->4)-beta-D glucans, (1-->6)-beta-D-glucan, CM-cellulose, insoluble (1-->3)-beta-D-glucans or aryl beta-D-glycosides. PMID- 8424791 TI - The linkage between binding of the C-terminal domain of hirudin and amidase activity in human alpha-thrombin. AB - A method derived from the analysis of viscosity effects on the hydrolysis of the amide substrates D-phenylalanylpipecolyl-arginine-p-nitroaniline, tosylglycylprolylarginine-p-nitroanaline and cyclohexylglycylalanylarginine-p nitroalanine by human alpha-thrombin was developed to dissect the Michaelis Menten parameters Km and kcat into the individual rate constants of the binding, acylation and deacylation reactions. This method was used to analyse the effect of the C-terminal hirudin (residues 54-65) [hir-(54-65)] domain on the binding and hydrolysis of the three substrates. The results showed that the C-terminal hir-(54-65) fragment affects only the acylation rate, which is increased approx. 1.2-fold for all the substrates. Analysis of the dependence of acylation rate constants on hirudin-fragment concentration, allowed the determination of the equilibrium binding constant of C-terminal hir-(54-65) (Kd approximately 0.7 microM). In addition this peptide was found to competitively inhibit thrombin fibrinogen interaction with a Ki which is in excellent agreement with the equilibrium constant derived from viscosity experiments. These results demonstrate that binding of hir-(54-65) to the fibrinogen recognition site of thrombin does not affect the equilibrium binding of amide substrates, but induces only a small increase in the acylation rate of the hydrolysis reaction. PMID- 8424792 TI - Bombesin stimulates distinct time-dependent changes in the sn-1,2-diradylglycerol molecular species profile from Swiss 3T3 fibroblasts as analysed by 3,5 dinitrobenzoyl derivatization and h.p.l.c. separation. AB - We have developed procedures for the analysis of endogenous diradylglycerol (DRG) molecular species using derivatization with 3,5-dinitrobenzoyl chloride. The introduction of this strong chromatophore enabled us to separate less than 1 nmol of DRG into its three classes (diacylglycerol, alkylacylglycerol and alkenylacylglycerol) using a combination of h.p.l.c. and t.l.c. followed by reversed-phase h.p.l.c. to resolve these classes into their component molecular species. When applied to Swiss 3T3 mouse fibroblasts stimulated with bombesin for 25 s, 5 min or 30 min, subtle time-dependent changes in the DRG patterns were observed, with only certain polyunsaturated 1,2-diacyglycerol species [18:0/20:3(n-9), 18:0/20:4(n-6), 18:0/20:4(n-3), 18:0/20:5(n-3), 18:1(n-9)/20:3(n 9), 18:1(n-9)/20:4(n-6), 16:0/22:6(n-3), 18:0/20:3(n-6) and 16:0/20:5(n-3)] showing significant agonist-stimulated increases. The amounts of the first six species were all raised at 25 s, whereas all except the latter two were elevated at 5 min. By 30 min these last species were also increased but 18:0/20:3(n-9) had returned to basal levels. Overall DRG levels, as measured by total molecular species peak area, remained effectively constant. No changes in the amount or species profile of 1-alkyl-2-acylglycerol were observed. Comparison of these species with the acyl-chain structure of phospholipids supports the idea that inositol lipids could be the source of DRG at early stimulation times, but phosphatidylcholine appears to be a phospholipase substrate at all times. These results indicate sequential activation of several phospholipases with different substrate specificities and/or access to different phospholipid pools. They also suggest that only polyunsaturated DRGs act as second messengers and that changes in the relative amounts of these species may trigger activation of different proteins and/or isoforms (e.g. the different isoforms of protein kinase C). PMID- 8424793 TI - Binding of human xanthine oxidase to sulphated glycosaminoglycans on the endothelial-cell surface. AB - Much evidence has suggested that the superoxide generated by xanthine oxidase (XOD) within the endothelial cell triggers characteristic free-radical-mediated tissue injuries. Although it has been reported that XOD exists not only in the cytoplasm, but also on the outside surface of the endothelial cell membrane, it is not clear how XOD localizes on the outside of the plasma membrane. Purified human xanthine oxidase (h-XOD) had an affinity for heparin-Sepharose. The binding was largely independent of the pH over the physiological range, whereas it tended to increase at lower pH and to decrease at higher pH. Exposure of h-XOD to the lysine-specific reagent trinitrobenzenesulphonic acid or the arginine-specific reagent phenylglyoxal caused it to lose its affinity for heparin-Sepharose. The binding of h-XOD to heparin is apparently of electrostatic nature, and both lysine and arginine residues are involved in the binding. h-XOD was found to bind to cultured porcine aortic endothelial cells, and this binding was inhibited by the addition of heparin or pretreatment of the cells with heparinase and/or heparitinase. Intravenous injection of heparin into two healthy persons led to a prompt increase in plasma h-XOD concentration. These results suggest that XOD localizes on the outside surface of endothelial cells by association with polysaccharide chains of heparin-like proteoglycans on the endothelial-cell membranes. Superoxide extracellularly generated by XOD may injure the source endothelial-cell membrane and also attract and activate closely appositional neutrophils, which themselves actually cause progressive oxidative damage. PMID- 8424794 TI - Order of uroporphyrinogen III decarboxylation on incubation of porphobilinogen and uroporphyrinogen III with erythrocyte uroporphyrinogen decarboxylase. AB - The isomeric compositions of the heptacarboxylic, hexacarboxylic and pentacarboxylic porphyrinogens formed by incubation of porphobilinogen with human red-cell haemolysates have been analysed and compared with those derived from incubation with chemically prepared uroporphyrinogen III as substrate. The results indicated that when supplied with an excess (3.7 microM) of exogenous uroporphyrinogen III, uroporphyrinogen decarboxylase utilized the substrate at random and a mixture of isomers was produced; whereas with uroporphyrinogen III generated enzymically from porphobilinogen as substrate a clockwise decarboxylation sequence was observed, resulting in the formation of intermediates mainly with the ring-D, rings-AD and rings-ABD acetate groups decarboxylated. Using [14C]uroporphyrinogen III as substrate at low concentrations (0.01-0.5 microM) also led to preferential decarboxylation of the ring-D acetate group. It was concluded that the order of uroporphyrinogen III decarboxylation is substrate-concentration-dependent, and under normal physiological conditions enzymic decarboxylation is most probably orderly and clockwise, starting at the ring-D acetate group. PMID- 8424795 TI - Site-directed mutation studies of human liver cytochrome P-450 isoenzymes in the CYP2C subfamily. AB - Evidence from human studies in vivo and in vitro strongly suggests that the methylhydroxylation of tolbutamide and the 4-hydroxylation of phenytoin, the major pathways in the elimination of these two drugs, are catalysed by the same cytochrome P-450 isoenzyme(s). In the present study we used site-directed mutagenesis and cDNA expression in COS cells to characterize in detail the kinetics of tolbutamide and phenytoin hydroxylations by seven CYP2C proteins (2C8, 2C9 and variants, and 2C10) in order to define the effects of small changes in amino acid sequences and the likely proteins responsible in the metabolism of these two drugs in man. Tolbutamide was hydroxylated to varying extents by all expressed cytochrome P-450 isoenzymes, although activity was much lower for the expressed 2C8 protein. While the apparent Km values for the 2C9/10 isoenzymes (71.6-131.7 microM) were comparable with the range of apparent Km values previously observed in human liver microsomes, the apparent Km for 2C8 (650.5 microM) was appreciably higher. The 2C8 enzyme also showed quite different sulphaphenazole inhibition characteristics. The 4-hydroxylation of phenytoin was also more efficiently catalysed by the 2C9/10 enzymes. These enzymes showed similarities in kinetics of phenytoin hydroxylation and sulphaphenazole inhibition compared with human liver phenytoin hydroxylase. Also of interest was the observation that, among the 2C9 variants, small differences in amino acid composition could appreciably affect both tolbutamide and phenytoin hydroxylations. The amino acid substitution Cys-144-->Arg increased both the rates of tolbutamide and phenytoin hydroxylations, while the Leu-359-->Ile change had a greater effect on phenytoin hydroxylation. We conclude that: (1) although 2C8 and 2C9/10 proteins metabolize tolbutamide. only 2C9/10 proteins play a major role in human liver; (2) 2C9/10 proteins also appear to be chiefly responsible for phenytoin hydroxylation; and (3) subtle differences in the amino acid composition of these 2C9/10 proteins can affect the functional specificities towards both tolbutamide and phenytoin. PMID- 8424796 TI - Non-electrostatic factors govern the hydrodynamic properties of articular cartilage proteoglycan. AB - The hydrodynamic frictional resistance to water flow exerted by articular cartilage proteoglycan is shown to be similar to that of proteoglycan isolated from Swarm rat chondrosarcoma, and independent of the state of aggregation of the proteoglycan. Frictional resistance is dependent, however, on the chain segments of the constituent chondroitin-sulphate and keratan-sulphate chains of the proteoglycan. Frictional resistance offered by chondroitin sulphate was independent of pH over the range 3.2-8.7. This confirms previous studies, associated with varying ionic strength and chemical modification of ionic groups of chondroitin sulphate, which showed that the frictional resistance offered by this molecule is independent of electrostatic factors. Water-structure-breaking and hydrogen-bond-breaking solvents were also without major effects on the flow resistance offered by chondroitin sulphate. An overall secondary structure of chondroitin sulphate was not evident, as it showed no significant difference to dextran in terms of its temperature dependence of relative viscosity. Local regions of rigid secondary structure, as manifested through inter-residue hydrogen bonding between sugar residues, is likely to control flow resistance as periodate-oxidized chondroitin sulphate and periodate-oxidized and reduced preparations showed a significant decrease in their frictional resistance to water. PMID- 8424797 TI - A mutation and an antibody that affect chemical cross-linking of low-density lipoprotein receptors on human fibroblasts. AB - Treatment of normal fibroblasts with the bifunctional cross-linking reagent DTSSP [3,3'-dithiobis(sulphosuccinimidylpropionate)] at 4 degrees C converted approximately 40% of the cell-surface low-density lipoprotein (LDL) receptors into a high-M(r) form, thought to represent receptor dimers. Preincubation of the cells with anti-(LDL receptor) monoclonal antibody 10A2 increased the proportion of surface receptors in the high-M(r) form after treatment with DTSSP at 4 degrees C to over 70%. Preincubation with LDL did not affect the proportion cross linked, but prevented the increase produced by antibody 10A2. Cross-linking at 37 degrees C was less efficient than at 4 degrees C and was not affected by preincubation with antibody 10A2. Surface LDL receptors on fibroblasts from the homozygous familial hypercholesterolaemic subject MM were not cross-linked by DTSSP, confirming that the mutation had produced a change in the conformation of the receptor molecule. Taken together, the results suggest that normal LDL receptors on at least one region of the surface membrane may be loosely associated in some form of multimeric array which alters its alignment differently in response to antibody 10A2 and to cooling. Mutations that alter the tertiary structure of the receptors could affect LDL binding by disturbing the arrangement of the array. PMID- 8424798 TI - Iodide modulation of the EDTA-induced iodine reductase activity of horseradish peroxidase by interaction at or near the EDTA-binding site. AB - Horseradish peroxidase (HRP) catalyses the reduction of iodinium ion (I+) to iodide by H2O2 in the presence of EDTA. I+ reduction occurs optimally at pH 6 whereas the enzyme catalyses iodide oxidation optimally at pH 3.5. Thus the two activities reside on the same enzyme with two characteristic pH optima. Iodide modulates the expression of the reductase activity by EDTA. Higher concentrations of iodide inhibit the reductase activity by EDTA. Nitrite, an electron donor, acts similarly to iodide. Both EDTA and nitrite competitively inhibit iodide oxidation, indicating that they compete with iodide for the same binding site for electron flow to the haem iron group. However, unlike iodide, EDTA converts compound I, not into the native enzyme, but into a compound absorbing at 416 nm which reduces I+ and then returns to the native form. The apparent equilibrium dissociation constant, KD, for the formation of the EDTA-HRP complex (15 mM) is doubled in the presence of iodide, indicating interference with EDTA binding by iodide. EDTA binds away from the haem iron centre and not through intramolecular Ca2+. The pH-dependence of EDTA binding indicates that an ionizable group of the enzyme with pKa 5.8, presumably a distal histidine, controls the binding. The data suggest that iodide competes with EDTA for compound I and modulates the iodine reductase activity by limiting the formation of the 416 nm-absorbing active compound. PMID- 8424799 TI - Multiple regulation of ornithine decarboxylase in enzyme-overproducing cells. AB - We have isolated from mouse FM3A cells a variant cell line, termed EXOD-1, that overproduces ornithine decarboxylase (ODC). The cells were resistant to alpha difluoromethylornithine, an irreversible inhibitor of the enzyme, and produced the enzyme protein to the extent of approx. 3-6% of total cytosolic protein. The rate of ODC synthesis in this cell line accounted for 25-50% of the rate of total protein synthesis. The amounts of the ODC gene and its mRNA in the variant cells were both about 60 times as much as those in wild-type FM3A cells. Upon removal of the inhibitor, the growth of the ODC-overproducing cells was stimulated approx. 2-fold. Under these conditions, the rate of ODC synthesis increased about 4-fold on day 1 and then decreased to near the original level by day 3. The amount of ODC mRNA increased about 1.7-fold on day 1 and 2.5-fold on day 3. No correlation was observed between changes in ODC synthesis rate and in ODC mRNA content, suggesting a translational repression of ODC mRNA due to accumulation of polyamines. In fact, the cellular contents of putrescine and spermidine markedly increased and that of spermine inversely decreased during the same period. Pulse chase experiments showed that the accumulation of putrescine and spermidine also elicited a rapid degradation of ODC. Excess amounts of newly synthesized putrescine and cadaverine were excreted into the medium, whereas spermidine, spermine and acetylated polyamines were undetectable there. We conclude that ODC regulation upon removal of the inhibitor is dependent on at least three steps, namely the level of mRNA, the translational efficiency of mRNA and the stability of the enzyme, the last two of which are involved in cellular polyamines. PMID- 8424800 TI - Domain organization of penicillin-binding protein 5 from Escherichia coli analysed by C-terminal truncation. AB - The structural organization of penicillin-binding protein (PBP) 5 was investigated by C-terminal truncation. Compared with other low-M(r) penicillin interacting proteins, PBP5 carries a C-terminal extension of about 100 amino acids. The sites for introduction of stop codons were chosen on the basis of the established three-dimensional structure of the Streptomyces albus G beta lactamase [Dideberg, Charlier, Wery, Dehottay, Dusart, Erpicum, Frere and Ghuysen (1987) Biochem. J. 245, 911-913] and comparative hydrophobic cluster analysis [Gaboriaud, Bissery, Bencheritt and Mornon (1987) FEBS Lett. 224, 149-155]. Two stop codons were introduced at positions Ile-354 or Val-348 to construct an optimized soluble form of PBP5 for crystallization purposes. The newly constructed soluble and enzymically active form (PBP5s353) was isolated by dye affinity chromatography and gave rise to small crystals. Another two stop codons were introduced at positions Arg-261 or Ala-276 to determine the minimal enzymically active 'core protein'. The truncated form (PBP5s275), missing the entire C-terminal extension, showed unaltered penicillin-binding characteristics and a catalytic-centre activity 40% that of PBP5s353 + 9 using bisacetyl-L-Lys-D Ala-D-Ala as substrate. This protein, however was more susceptible to proteolytic degradation, which might indicate a role of the C-terminal portion in stabilizing the protein. PMID- 8424801 TI - The competition plot: a simple test of whether two reactions occur at the same active site. AB - The competition plot is a method for determining whether or not two enzyme catalysed reactions occur at the same active site. It is a plot of total rate against p, where p varies from 0 to 1 and specifies the concentrations (1-p)a0 and pb0 of two substrates in terms of reference concentrations a0 and b0 chosen so as to give the same rates at p = 0 and p = 1. If the two substrates react at the same site, the competition plot gives a horizontal straight line, i.e. the total rate is independent of p. Independent reactions at two separate sites give a curve with a maximum; separate reactions with cross-inhibition generate curves with either maxima or minima according to whether the Michaelis constants of the two substrates are smaller or larger than their inhibition constants in the other reactions. Although ambiguous results can sometimes arise, experimental strategies exist for avoiding them, for example working as close as possible to the lower of the two limiting rates. When tested with yeast hexokinase, the plot indicated phosphorylation of glucose and fructose at the same site. Conversely, with a mixture of yeast hexokinase and galactokinase it indicated phosphorylation of glucose and galactose at different sites. In both cases the observed behaviour agreed with the known properties of the enzymes. A slight modification to the definition of this plot allows it to be applied also to enzymes that deviate from Michaelis-Menten kinetics. PMID- 8424802 TI - Sequence-specific binding of [N-MeCys3,N-MeCys7]TANDEM to TpA. AB - The sequence selective binding of [N-MeCys3,N-MeCys7]TANDEM to DNA has been studied by footprinting experiments on DNA fragments containing the self complementary sequences CGCGATATCGCG, CGCGTATACGCG, CGCGTTAACGCG and CGCGAATTCGCG. DNAase I and micrococcal nuclease reveal drug-induced footprints with the central sequences ATAT, TATA and TTAA, but not AATT, suggesting that the ligand binds to the dinucleotide TpA. The ligand renders certain adenines hyper reactive to diethyl pyrocarbonate. These are observed with ATAT, TATA and TTAA, but not AATT, and are located both within, and distal to, the TpA-binding sites. PMID- 8424804 TI - Influence of human recombinant interleukin-1 beta on the enantioselective disposition of propranolol in rats. AB - The influence of i.v. administration of 10 micrograms/kg recombinant human interleukin-1 beta (rhIL-1 beta), a putative mediator of inflammation, on the pharmacokinetics and metabolism of the propranolol enantiomers was studied in rats aged 3, 12 and 24 months. After oral administration of rac-propranolol to control rats of the three age groups, the plasma concentrations of (R) propranolol were higher than those of (S)-propranolol. Administration of IL-1 beta increased the plasma concentrations of the (R)-enantiomer markedly and significantly, those of the (S)-enantiomer only to a lesser degree. For both enantiomers an important increase in plasma binding was found in the IL-1 beta treated rats, which was linked to the increase in alpha 1-acid glycoprotein levels. The in vitro clearance, measured in 3-month-old rats using the 9000 g liver fraction, was for neither of the propranolol enantiomers influenced by IL-1 beta treatment, which is in keeping with the unchanged cytochrome P450 content. The enantioselective influence of IL-1 beta treatment on the pharmacokinetics of propranolol was also present in 12- and 24-month-old rats, although somewhat less pronounced in the latter group. Our results show an enantioselective influence of IL-1 beta treatment on the pharmacokinetics of propranolol in the rat, favouring the (R)-enantiomer. PMID- 8424803 TI - Human fetoacinar pancreatic protein: an oncofetal glycoform of the normally secreted pancreatic bile-salt-dependent lipase. AB - A fetoacinar pancreatic protein (FAP) associated with the ontogenesis, differentiation and oncogenic transformation of the human exocrine pancreas has been purified from pancreatic juices of patients suffering from pancreatitis or duodenal cancers invading the pancreas [Escribano and Imperial (1989) J. Biol. Chem. 264, 21865-21871]. This protein has striking similarities, i.e. M(r), amino acid composition and N-terminal sequence, to the bile-salt-dependent lipase (BSDL) of normal human pancreatic secretion. The aim of this study was to gain further insight into the nature of the two proteins. Reactivity with the mouse monoclonal antibody J28 (mAb J28), which characterizes FAP, and enzyme activity could not be dissociated during biochemical purification of BSDL. Furthermore, a polyclonal antiserum raised against purified human BSDL reacted completely with FAP in Western-blot analysis giving additional support to the idea of similar molecular structures for BSDL and FAP. However, by the same technique, mAb J28 reacted with a relatively restricted population of BSDL molecules. The classical BSDL preparation could be separated into molecules bearing the J28 epitope and those devoid of it by immunoaffinity on immobilized mAb J28. The two subpopulations had identical N-terminal sequences and some differences in their amino acid compositions. However, they had different carbohydrate compositions. J28-epitope-bearing molecules were active on BSDL substrates, although their specific activity was decreased. These results are consistent with the existence of two closely related polypeptide chains with different glycan counterparts. Therefore, if the name FAP is reserved for molecules bearing the J28 epitope, which is linked to a carbohydrate-dependent structure. FAP could represent an oncofetal-related variant of BSDL. Our result is the first demonstration of the existence of an oncofetal-type subpopulation of an otherwise normally secreted human pancreatic enzyme. PMID- 8424805 TI - Hepatic cytochrome P450 induction in goats. Effects of model inducers on the metabolism of alkoxyresorufins, testosterone and ethylmorphine, and on apoprotein and mRNA levels. AB - Male and female African dwarf goats were treated orally with phenobarbital (PB) or triacetyloleandomycin (TAO), or subcutaneously with beta-naphthoflavone (BNF). Hepatic microsomal cytochrome P450 content was increased by PB and TAO, but not by BNF. PB effects on P450 activities were non-selective: ethoxyresorufin deethylase (EROD) and pentoxyresorufin depentylase (PROD), hydroxylation of testosterone (TST) and demethylation of ethylmorphine (ETM) were all induced by a factor of 2-3. A similar non-selective induction was observed with TAO, except for EROD and PROD (no effects). After PB and TAO treatment, increased levels of a protein cross-reactive with anti-sheep P450 3A and 2B were found. Thus, in dwarf goats, both PB and TAO appeared to be P450 3A inducers. Selective PB effects related to a P450 2B form on PROD are lacking but 16 alpha-hydroxylation of TST was induced markedly. At the mRNA level, PB induced an mRNA that showed good sequence homology with a human P450 3A4 cDNA probe, rather than with a rat 3A1 probe. BNF selectively induced EROD, whereas TST hydroxylation and ETM dealkylation were inhibited. With BNF-treated animals, increased concentrations of a protein cross-reactive with anti-rat P450 1A1/1A2 and of an mRNA that showed homology with a human 1A1 cDNA probe, but not with a mouse 1A1/1A2 probe, were observed. PMID- 8424806 TI - Antioxidant and iron-chelating activities of the flavonoids catechin, quercetin and diosmetin on iron-loaded rat hepatocyte cultures. AB - The cytoprotective effect of three flavonoids, catechin, quercetin and diosmetin, was investigated on iron-loaded hepatocyte cultures, considering two parameters: the prevention of iron-increased lipid peroxidation and the inhibition of intracellular enzyme release. These two criteria of cytoprotection allowed the calculation of mean inhibitory concentrations (IC50) which revealed that the effectiveness of these flavonoids could be classified as follows: catechin > quercetin > diosmetin. These IC50 values have been related to structural characteristics of the flavonoids tested. Moreover, the investigation of the capacity of these flavonoids to remove iron from iron-loaded hepatocytes revealed a good relationship between this iron-chelating ability and the cytoprotective effect. The cytoprotective activity of catechin, quercetin and diosmetin could thus be ascribed to their widely known antiradical property but also to their iron-chelating effectiveness. These findings increase further the prospects for the development and clinical application of these potent antioxidants. PMID- 8424807 TI - Effect of sodium benzoate on cerebral and hepatic energy metabolites in spf mice with congenital hyperammonemia. AB - The sparse-fur (spf) mutant mouse has an X-linked deficiency of ornithine transcarbamylase and develops congenital hyperammonemia similar to that seen in human patients. We studied the effect of sodium benzoate (2.5, 5 and 10 mmol/kg body wt) on ammonia, glutamine and glutamate, as well as various intermediates of energy metabolism in brain and liver of normal CD-1/Y and hyperammonemic spf/Y mice. The ammonia concentration of brain was decreased with 2.5 mmol sodium benzoate in spf/Y mice, whereas higher doses resulted in a significant increase in both liver and brain. Cerebral glutamine content decreased generally in a dose dependent manner, both in normal and affected mice, following treatment with various doses of sodium benzoate. Cerebral glutamate concentrations were increased only in spf mice treated with sodium benzoate, whereas ATP and acetyl CoA were decreased (P < 0.001), in both normal and affected mice, indicating that glutamine synthesis may be affected by ATP availability. Free CoA levels were decreased (P < 0.05) only in liver in both groups of treated mice, whereas pyruvate concentrations were elevated (P < 0.05) in affected mice following sodium benzoate administration. The results demonstrate that a dose of 2.5 mmol sodium benzoate/kg body wt has a beneficial effect in reducing cerebral ammonia with a concomitant decrease in glutamine. However, the results suggest that many of the metabolite changes observed following higher doses of benzoate could be due to depletion of ATP, free CoA and acetyl CoA levels, possibly secondary to benzoyl CoA accumulation. The response of the spf/Y mouse to sodium benzoate was different from that of the control CD-1/Y mouse, which could be due to its urea cycle dysfunction and a chronic hyperammonemic state. Hence, the spf/Y mouse may be the ideal animal model for studying the pharmacology of sodium benzoate in hyperammonemic disorders at both the cerebral and hepatic levels. PMID- 8424808 TI - Intracellular distribution of various boron compounds for use in boron neutron capture therapy. AB - The neutron capture reaction in boron (10B(n, alpha)7Li) generates two short range particles with high linear energy transfer. The effect of neutron capture therapy depends on the selective localization of 10B atoms in target cells. The determination of the distribution of boron compounds in cancer cells at the subcellular level is required for the understanding of the effect of this treatment. The monomeric sulfhydryl borane (BSH) compound has been used clinically in Japan and preclinically in the U.S.A. Recently, new compounds have been developed: a dimeric sulfhydryl borane (BSSB), a boronophenylalanine (BPA), and two porphyrin complexes (BOPP and VCDP). This study demonstrates that the porphyrin complexes (BOPP and VCDP) are more cytotoxic than the other three compounds to the rat 9L gliosarcoma cell line. Using atomic absorption spectrophotometry to determine boron content for cellular uptake studies of these agents, we found that of the five compounds tested BOPP (25 microM) exposure resulted in the greatest boron uptake averaging 305 ng B/10(6) cells. BSSB (500 microM) was second averaging 93 ng B/10(6) cells, BSH (500 microM) third averaging 62 ng B/10(6) cells, VCDP (25 microM) fourth averaging 58 ng B/10(6) cells, and BPA (500 microM) fifth averaging 7.4 ng B/10(6) cells. Data on the distribution of boron in the nuclei, mitochondria, lysosomes, microsomes, and cytosomes of 9L cells are also presented. PMID- 8424809 TI - Relationship between nitroglycerin-induced vascular relaxation and nitric oxide production. Probes with inhibitors and tolerance development. AB - We have shown previously that nitric oxide (NO) is generated from nitroglycerin (NTG) through enzyme-mediated reactions in the bovine coronary artery smooth muscle cell, but it is not known whether this metabolic conversion plays a significant role in the pharmacologic action of NTG, viz. relaxation. In this study, we developed a technique that allowed direct measurement of NO from intact bovine coronary arterial rings that were incubated previously with NTG, and examined whether changes in NTG-induced relaxation were accompanied by parallel changes in NO generation. Co-incubation of the vascular preparations with a potent inhibitor of glutathione-S-transferases (GSTs), bromosulfophthalein (up to 200 microM), did not affect NTG-induced relaxation, nor did it alter NO generation from NTG in the preparation. In contrast, 1-chloro-2,4-dinitrobenzene (CDNB), a GST substrate, inhibited NO generation as well as the relaxation response of NTG in the intact vascular tissue preparation. CDNB, however, did not decrease the relaxant responses of nifedipine and isoproterenol. Thus, the inhibitory effect of CDNB on NTG-induced relaxation and NO production appeared specific. When bovine coronary rings were made tolerant to NTG by pretreatment with 0.44 mM NTG for 1 hr, the EC50 was shifted to the right 162-fold, and NO generation was also reduced in intact rings and tissue homogenates. However, when the homogenates were further subfractionated to microsomes and cytosols, or when homogenates were allowed to stand for a similar time period necessary for subfractionation, the difference in NO production from control versus tolerant tissue preparations disappeared. It is possible, therefore, that the NTG-induced tolerance process might have been partially reversed during this time period. Results of this study identified CDNB as an apparently specific inhibitor of NTG action, but showed that GST-mediated reactions were probably not involved in the metabolic activation of NTG. Our results also indicated that tissue NO generation from NTG was positively related to the relaxation responses generated by this nitrovasodilator. PMID- 8424810 TI - Species-dependent differences in the biochemical effects and metabolism of 5 benzylacyclouridine. AB - The pharmacokinetics and biochemical effects of the uridine phosphorylase (UrdPase) inhibitor 5-benzylacyclouridine (BAU) were investigated in the mouse, rat and monkey. Following i.p. administration of BAU (30 mg/kg) in the mouse and i.v. administration in the rat and monkey, initial BAU plasma half-life values were 36, 36 and 25 min, and the areas under the plasma BAU concentration versus time curves (AUC) were 127, 80 and 76 microM.hr, respectively. Rats were also dosed p.o. and i.v. with BAU at 90 mg/kg, and a comparison of the AUC values showed an oral bioavailability of 70%. Analyses of plasma samples by HPLC indicated that the metabolism of BAU differed in these species. A major BAU metabolite was observed in monkeys. Its concentration was greater than or equal to that of BAU in almost every plasma sample, and its elimination paralleled that of BAU. Urinary recovery of the metabolite was 10-fold higher than the recovery of unchanged drug. The compound was identified as the ether glucuronide of BAU by its UV absorption spectrum, its co-elution with BAU after incubation with beta glucuronidase, and liquid chromatography/mass spectrum analysis. A different metabolite was detected in rat plasma; its maximum concentration was 15% of the BAU level, and its elution position on the HPLC chromatogram was not affected by the action of beta-glucuronidase. BAU had equivalent potency against UrdPase in liver extracts from the three species, with Ki values of about 0.17 microM. However, the in vivo effects of BAU on plasma uridine concentrations were species dependent. In mice, a 30 mg/kg i.p. dose of BAU increased the plasma uridine concentration to 11 microM from a control level of 1.8 microM. In the rat, a 30 mg/kg i.v. dose of BAU increased plasma uridine to 2.1 from 1.1 microM control levels, and a 300 mg/kg oral dose resulted in a peak plasma uridine concentration of only 6 microM. In the monkey, BAU (30 mg/kg, i.v.) had no effect on plasma uridine despite the presence of 10-100 microM BAU levels in plasma for 1.5 hr. These data show that there are significant differences in the biochemical effects and metabolism of BAU in CD-1 mice, CD rats and cynomolgus monkeys. PMID- 8424811 TI - Chloroquine stabilization of phospholipid membranes against diacylglycerol induced perturbation. AB - The effects of 1-stearoyl,2-sn-arachidonoylglycerol (SAG) and the antimalarial drug chloroquine on lipid bilayer structure were studied by 2H-NMR spectroscopy. Model lipid systems were established with compositions similar to those of normal human erythrocytes, malaria-infected erythrocytes, or malaria parasite membranes. The 2H-NMR spectra of the membranes formed from the lipids extracted from normal human erythrocytes were similar to those obtained using the corresponding lipid mixtures. The order parameters of the model "infected" and model "parasite" membranes were reduced markedly relative to that of normal erythrocytes. Addition of SAG induced formation of non-bilayer lipid phases in all lipid systems. Only a small decrease in the order parameters of the acyl side chains of the phosphatidylserine, but not of the phosphatidylcholine component of the lipid membranes, was observed upon the addition of chloroquine. A dramatic effect was observed upon the addition of chloroquine to the SAG-containing membranes: this antimalarial almost totally abolished the formation of SAG-induced non-bilayer lipid phases. Since SAG, endogenously formed in erythrocyte membranes, is a potent activator of phospholipase A2, this membrane-stabilizing action of chloroquine may partially account for the phospholipase A2-inhibiting properties of this drug, and, consequently, for both its therapeutic and toxic modes of action. PMID- 8424812 TI - Hypoxia and oxygen dependence of cytotoxicity in renal proximal tubular and distal tubular cells. AB - Ischemia and hypoxia are major causes of renal failure and altered oxygen supply may affect renal responses to toxic chemicals. In vitro experiments were designed to evaluate the susceptibility of isolated proximal tubular (PT) and distal tubular (DT) cells from rat kidney to brief periods of oxygen deprivation and to assess how variations in oxygen supply affect chemical-induced cytotoxicity. Isolated cells were incubated for 1 hr in either oxygen (95% O2/5% CO2), air (21% O2), or nitrogen (95% N2/5% CO2) atmosphere. PT cells exhibited no injury due to brief oxygen deprivation whereas DT cells exhibited moderate, but significant injury, indicating that DT cells are more susceptible than PT cells to hypoxic injury. The cytotoxicity of chemicals that alter cellular redox status [i.e. tert butyl hydroperoxide (tBH), menadione, methyl vinyl ketone] and the cytotoxicity of "chemical hypoxia" [i.e. KCN + iodoacetic acid] were greatest in air, intermediate in oxygen, and lowest in nitrogen. In contrast, the cytotoxicity of the alkylating agent N-dimethylnitrosamine was independent of oxygen concentration and the cytotoxicity of p-aminophenol was related directly to oxygen concentration. The mechanism of the oxygen dependence of chemical injury was investigated further, employing tBH as a model toxicant. tBH metabolism was oxygen independent in both PT and DT cells. Depletion of cellular protein sulfhydryl groups by tBH increased with increasing oxygen concentration and lipid peroxidation due to tBH was inhibited in nitrogen but was not different in air as compared with oxygen. Although these processes may contribute to the much lower toxicity in nitrogen as compared with oxygen, it does not explain the higher toxicity in air as compared with that in oxygen. Other processes that predominate at lower oxygen concentrations but that only produce injury if enough oxygen is present are likely to be responsible for the enhanced susceptibility of both PT and DT cells to oxidants in air as compared with oxygen. PMID- 8424813 TI - Interleukin-6 as a mediator responsible for inflammation-induced increase in plasma angiotensinogen. AB - The concentration of plasma angiotensinogen increases upon induction of inflammation. Studies were carried out using serum samples collected from mice and rats after injection of lipopolysaccharide (LPS) to determine whether interleukin-6 (IL-6) is a mediator responsible for the inflammation-induced increase of angiotensinogen synthesis in liver cells. Serum collected from mice or rats 2 and 4 hr after injection of LPS contained a factor that stimulated [35S]methionine incorporation into angiotensinogen newly synthesized by rat hepatoma H4IIEC3 (H4) cells. Assay of IL-6 using an IL-6-dependent murine hybridoma, MH60.BSF2 cells, showed the presence of IL-6-like activity in sera of mice or rats 2 and 4 hr after injection of LPS. Anti-mouse IL-6 monoclonal antibody completely inhibited not only the IL-6-like activity present in LPS treated mouse serum but also the ability of the serum to stimulate angiotensinogen synthesis of H4 cells. These results suggest that increased synthesis of angiotensinogen in the liver after induction of inflammation is mediated by IL-6, a cytokine important in immune reactions and the hepatic acute phase response. PMID- 8424814 TI - Induction of CYP2B1 and 3A1, and associated monoxygenase activities by tamoxifen and certain analogues in the livers of female rats and mice. AB - Previous studies suggest long-term feeding of tamoxifen (Z-1-[4-(2-dimethylamino ethoxy)phenyl]1,2-diphenyl-1-butane) to rats gives rise to liver tumours, while mice are resistant. The effects of tamoxifen on cytochrome P450 isoenzymes and associated monoxygenase activities in the livers of female Fischer rats and C57Bl/6 and DBA/2 mice have been compared. Total microsomal cytochrome P450 was not induced in the livers of rats given tamoxifen (45 mg/kg daily for 4 days) and was in fact significantly reduced after 3 days treatment. In contrast, there was a 30-60-fold increase in the metabolism of benzyloxy- and pentoxyresorufins to resorufin. Ethoxyresorufin O-deethylase was induced only 2.5-fold. The regio- and stereo-specific hydroxylation of testosterone following tamoxifen pretreatment of rats showed a general time- and dose-dependent induction. 6 beta- and 16 alpha hydroxylation of testosterone together with oxidation to androstenedione were increased 2-3-fold while 2 beta-hydroxylation was induced only marginally, suggesting that tamoxifen produces a mixed pattern of induction with a significant phenobarbitone-like component. No induction of the 2 beta- or 6 beta hydroxylation pathway occurred in either mouse strain. In rats, immunoblotting experiments with polyclonal antibodies raised against CYP2B1 or 3A1 showed that tamoxifen pretreatment resulted in 2-3-fold increases in both CYP2B1, 2B2 and 3A1 proteins, relative to controls. Immunohistochemistry of rat liver sections showed a centrilobular localization of these induced proteins. Similar patterns of induction as measured by immunoblotting experiments and testosterone hydroxylation were seen following the administration of structurally related analogues, toremifene and droloxifene (3-hydroxytamoxifen), thought to be non carcinogenic in the rat. No induction of these monooxygenase activities was seen in C57Bl/6 mice and only small increases in benzyloxy and pentoxyresorufin metabolism were in DBA/2 mice. It is suggested that the induction of cytochrome P450-dependent activities by tamoxifen may result in accelerated liver metabolism of this drug with important implications for the disposition of tamoxifen in vivo and also for its metabolic conversion to genotoxic metabolite(s). The difference in inducibility of cytochrome P450-dependent monooxygenase activities between rats and mice offers a plausible and testable hypothesis that the difference in tamoxifen metabolism between the two species may contribute to their carcinogenic response to tamoxifen. PMID- 8424815 TI - Channel catfish liver monooxygenases. Immunological characterization of constitutive cytochromes P450 and the absence of active flavin-containing monooxygenases. AB - Multiple drugs and pesticides are used in the aquaculture of channel catfish in the Southeastern United States. However, little is known regarding the enzymatic metabolism of these chemicals in the fish. Western blots, utilizing polyclonal antibodies raised against five purified rainbow trout liver cytochrome P450 enzymes, revealed at least two protein bands that were approximately 50 kDa (CATL 1) and 53 kDa (CATL-2). Anti-trout LMC3 and LMC4 only hybridized with the 53 kDa protein, whereas anti-trout LMC1, LMC2, and LMC5 recognized both proteins. Cytochrome P450-catalyzed activities (testosterone and progesterone hydroxylases) associated with LMC1 and LMC5 were also found in catfish liver microsomes. These data suggest that at least two constitutive forms of cytochrome P450 are present in the liver of juvenile channel catfish. Western blots utilizing antibodies raised against rabbit-lung flavin-containing monooxygenases (FMO) showed hybridization with two proteins from rainbow trout liver microsomes, but no cross reaction with microsomes from catfish liver. N,N,-Dimethylaniline N-oxidase and methimazole oxidase were observed in microsomes from trout, but were absent in catfish liver microsomes prepared in three different laboratories. Consequently, FMO do not appear to be present in liver microsomes from channel catfish or they are rapidly degraded during tissue homogenization. PMID- 8424816 TI - Sensitivity of aldehyde dehydrogenases in murine tumor and hematopoietic progenitor cells to inhibition by chloral hydrate as determined by the ability of chloral hydrate to potentiate the cytotoxic action of mafosfamide. AB - Several murine aldehyde dehydrogenases, most notably AHD-2, are known to catalyze the detoxification of cyclophosphamide, mafosfamide, and other oxazaphosphorines. Thus, cellular sensitivity to these agents decreases as the relevant aldehyde dehydrogenase activity increases, and vice versa. Chloral hydrate is a sedative/hypnotic agent that is sometimes administered to patients being treated with cyclophosphamide. It is known to inhibit some, but not all, aldehyde dehydrogenases. Murine (CFU-S, CFU-GEMM and CFU-Mk) and human (CFU-Mix, CFU-GM, BFU-E and CFU-Mk) hematopoietic progenitor cells, as well as murine oxazaphosphorine-resistant (L1210/OAP and P388/CLA) tumor cells, are known to contain the relevant aldehyde dehydrogenase activity but the identity of the specific enzyme present in the normal cells is unknown and may be different than that, namely AHD-2, present in neoplastic cells. In that event, the potential exists to inhibit the detoxification of the oxazaphosphorines in tumor cells without inhibiting this event in normal cells; the net effect of such a selective inhibition would be to increase the margin of safety of the oxazaphosphorines. In ex vivo experiments, chloral hydrate markedly potentiated the antitumor activity of mafosfamide against oxazaphosphorine-resistant L1210/OAP and P388/CLA cells. It did not potentiate the cytotoxic action of mafosfamide against any of the murine or human hematopoietic cells tested, even at concentrations which fully restored the sensitivity of the resistant tumor cell lines to this agent. One explanation for these observations is that hematopoietic progenitor, and the resistant tumor, cells express different relevant aldehyde dehydrogenases and that these aldehyde dehydrogenases differ in their sensitivity to inhibition by chloral hydrate. Consistent with this notion were the observations that AHD-2 was exquisitely sensitive to inhibition by chloral hydrate, whereas two other aldehyde dehydrogenases that also catalyze the detoxification of aldophosphamide, namely AHD-12a, b and AHD-13, were relatively unaffected. PMID- 8424817 TI - The role of metallothionein, glutathione, glutathione S-transferases and DNA repair in resistance to platinum drugs in a series of L1210 cell lines made resistant to anticancer platinum agents. AB - The glutathione contents, glutathione S-transferase activities and metallothionein contents have been measured in a series of L1210 cell lines which show decreased sensitivities to platinum drugs. Resistance to cisplatinum cisDDP, cis-diamminedichloroplatinum (II)] and chip [ioproplatin, cisdichloro-bis isopropylamine-trans dihydroxy platinum IV] was found to correlate with glutathione levels but not metallothionein. Conversely, resistance to tetraplatin was found to be correlated with metallothionein but not glutathione levels. However, depletion of glutathione by buthionine 1-sulphoximine sensitizes all cell lines to the effects of cisDDP, chip and tetraplatin [d,1-trans-tetrachloro 1,2-diamino-cyclohexanplatinum (IV)]. Inhibition of DNA repair by aphidicholin or caffeine also partially restored sensitivity to these platinum drugs. These results indicate the complexity of the changes occurring upon the development of drug resistance. PMID- 8424818 TI - Relationship between peroxisome-proliferating sulfur-substituted fatty acid analogs, hepatic lipid peroxidation and hydrogen peroxide metabolism. AB - The effect of the administration of three peroxisome-proliferating sulfur substituted fatty acid analogs on hepatic antioxidant status and lipid peroxidation was studied in rats. After 14 days of treatment, the ratio of induction of peroxisomal fatty acyl-CoA oxidase to catalase was 4.2 and 3.5 in rats treated with 1,10 bis-(carboxymethylthio)decane (BCMTD) and 1-mono (carboxymethylthio)tetradecane (CMTTD), respectively, while the corresponding ratio was 1.3 in 1-mono (carboxyethylthio)tetradecane (CETTD)-treated rats. As compared to the controls an increase in hepatic hydrogen peroxide content was noted in BCMTD- and CMTTD-treated rats, but not CETTD-treated rats. Hepatic lipid peroxidation was increased in all the three treatment groups in a manner not related to the potency of the compounds to induce the peroxisomal hydrogen peroxide metabolizing enzymes. Hepatic glutathione content increased while the activities of its associated enzymes such as glutathione transferase, glutathione peroxidase and glutathione reductase decreased in all the treated rats. Taken together, our data show a relationship between the levels of hydrogen peroxide and lipid peroxidation in rat livers treated with BCMTD and CMTTD. However, increased hepatic lipid peroxidation in CETTD-treated rats cannot be accounted for by the changes in the peroxisomal enzymes. PMID- 8424819 TI - Biotransformation of glyceryl trinitrate by rat aortic cytochrome P450. AB - Denitration of glyceryl trinitrate (GTN) by the microsomal fraction of rat aorta was found to be NADPH dependent and followed apparent first-order kinetics (T1/2 70.1 min). Biotransformation of GTN was regioselective for glyceryl-1,2-dinitrate formation, and was inhibited by carbon monoxide, SKF-525A, and oxygen. In aortic microsomes prepared from phenobarbital-pretreated rats, biotransformation was increased 7-fold, and was regioselective for glyceryl-1,3-dinitrate formation. These data strongly suggest the involvement of aortic cytochrome P450 in the biotransformation of GTN. PMID- 8424820 TI - Nature and role of xenobiotic metabolizing esterases in rat liver, lung, skin and blood. AB - In the present study, the distribution and nature of esterases in the rat which hydrolysed fluazifop-butyl, carbaryl, paraoxon and phenylacetate were investigated. Vmax and Km values for the hydrolysis reactions were determined. Fluazifop-butyl was hydrolysed to fluazifop by rat liver (Vmax mumol/min/g microsomes 6.2 +/- 0.4; cytosol 6.84 +/- 0.85), lung (Vmax microsomes 0.38 +/- 0.1; cytosol 1.5 +/- 0.32) and skin (Vmax microsomes 0.02 +/- 0.0015; cytosol 0.4 +/- 0.06) and by plasma (Vmax mumol/min/mL 5.8 +/- 0.48) and red blood cells (Vmax 0.03 +/- 0.015). Significant inhibition by paraoxon and bismitrophenol phosphate indicated the involvement of carboxylesterases. Carbaryl was hydrolysed by liver, lung and skin at a lower rate by microsomal fractions (Vmax nmol/min/g 2.1 +/- 0.25, 1.6 +/- 0.25, 0.2 +/- 0.035, respectively) compared to cytosolic fractions (Vmax 6.7 +/- 0.75, 1.4 +/- 0.36, 0.5 +/- 0.12) and plasma (Vmax nmol/min/mL 3.0 +/- 0.25). Hydrolysis involved carboxylesterases. Paraoxon was hydrolysed by paraoxonases/arylesterases only in the plasma (Vmax nmol/min/mL 246 +/- 12) and microsomal fractions from liver (Vmax 330 nmol/min/g +/- 25) and lung (Vmax 2 +/- 0.25). Phenylacetate was hydrolysed by both microsomal and cytosolic fractions from all tissues studied. Hydrolysis involved arylesterases in the microsomes and carboxylesterases in the cytosol. Extrahepatic hydrolysis may be important following some routes of exposure to xenobiotic esters. PMID- 8424821 TI - Oxidative stress-induced activation of microsomal glutathione S-transferase in isolated rat liver. AB - The activation of microsomal glutathione (GSH) S-transferase in isolated rat liver by oxidative stress was investigated using both ischemia/reperfusion and perfusion with hydrogen peroxide. When the isolated liver was reperfused for 30 min and 60 min after 90 min ischemia, microsomal GSH S-transferase activity, but not cytosolic transferase activity, was increased 1.2-fold and 1.3-fold, respectively. In addition, microsomal GSH peroxidase activity was also significantly increased after 60 min reperfusion following ischemia. The increase in microsomal GSH S-transferase activity by ischemia/reperfusion was reversed by dithiothreitol. When N-ethylmaleimide, which activates microsomal GSH S transferase by covalent binding to the cysteine residue of the enzyme, was incubated with microsomes, transferase activity was increased to 526% in control microsomes and to 399% in liver that underwent ischemia/reperfusion liver. These data indicate that microsomal GSH S-transferase is activated by ischemia/reperfusion of the liver by means of disulfide bond formation. When rats were pretreated with a catalase inhibitor 3-amino-1,2,4-triazole for 6 weeks, microsomal GSH S-transferase activity was increased 1.4-fold by ischemia/reperfusion, corresponding to a 1.8-fold increase as compared to the non perfused liver of untreated rats. Perfusion of the isolated liver with hydrogen peroxide (1 mM, 15 min) also caused a significant increase in microsomal GSH S transferase activity with a concomitant decrease in GSH content, confirming that liver microsomal GSH S-transferase in rats was activated in vivo by oxidative stress. PMID- 8424822 TI - Effects of niacin on biliary lipid output in the rat. AB - The mechanisms for the hypocholesterolaemic action of niacin (nicotinic acid) were examined in rats administered niacin at a dose of 400 mg/kg body wt/day for either 2 or 4 weeks. Another group of rats were administered diosgenin, an inhibitor of acyl-CoA:cholesterol acyltransferase, as a 1% (w/w) supplement in the diet for 7 days. Both agents produced small increases in bile flow rates (up to 40%) and mild hepatotoxicity evidenced by small increases in serum transaminase activities. Niacin treatment for 2 or 4 weeks lowered serum cholesterol concentrations by 13% or 29%, respectively, with the greatest decrease occurring in the low density lipoprotein fraction. This was accompanied by relatively large increases in biliary cholesterol output (114% and 130% after 2 and 4 weeks treatment, respectively) with smaller increases in the biliary output of phospholipid (18% and 45%) and bile acid (26% and 14%). Diosgenin treatment increased serum cholesterol by 29% and increased the biliary output of cholesterol, phospholipid and bile acid by 800%, 10% and 45%, respectively. Thus, both agents increased the cholesterol saturation of bile (100% by niacin, 500% by diosgenin). Cholesterol and phospholipid in fistula bile from control rats were present in lamellar and micellar forms. Niacin treatment did not alter the physical form of biliary lipids whilst diosgenin caused the appearance of vesicular lipid in fistula bile. Thus, increased biliary secretion of cholesterol explains, at least in part, the hypocholesterolaemic action of niacin. In addition, since aggregation of biliary vesicles is involved in cholesterol gallstone formation in humans, the non-appearance of vesicular material in fistula bile from niacin-treated rats may be of some importance. PMID- 8424823 TI - Characterization of specific cytochrome P450 enzymes responsible for the metabolism of diazepam in hepatic microsomes of adult male rats. AB - The role of several P450 enzymes in the metabolism of diazepam (DZ) has been investigated. Hepatic microsomes of adult male rats were pretreated with antisera raised against the P450s CYP3A2, 2B1, 2C6, 2C11, 2D1 and 2E1, and their influence on the subsequent metabolism of DZ was determined by simultaneously measuring the changes in the relative rates of formation of its metabolites. Several forms of P450 were found to be positively involved in DZ metabolism. Antisera of the "male specific" P450 enzyme CYP2C11 partially inhibited both DZ N-demethylase and C3 hydroxylase activities (60%) which resulted in decreased formations of N desmethyl-DZ (NDZ) and 3-hydroxy-DZ (3HDZ), respectively. In a reconstitution experiment with the purified enzyme, CYP2C11 predominantly catalysed the formation of NDZ from DZ. Antisera of a further male-specific P450 CYP3A2 strongly inhibited (95%) the C3 hydroxylase of DZ and thus 3HDZ formation. A corresponding reconstitution experiment with this same P450 enzyme gave 3HDZ as principal product. CYP2D1 antisera inhibited the aromatic hydroxylation of DZ (98%) and subsequent formation of 4'-hydroxy-DZ (4'HDZ). This enzyme was also observed to inhibit DZ N-demethylase activity (60%). A reconstitution experiment with pure CYP2D1 catalysed the formation of both 4'HDZ and NDZ. PMID- 8424824 TI - The modification of low density lipoprotein by the flavonoids myricetin and gossypetin. AB - Myricetin and gossypetin, two hexahydroxylated flavonoids, are capable of modifying low density lipoprotein (LDL) to increase greatly its uptake by macrophages. When human 125I-labelled LDL was incubated with 100-1000 microM myricetin or gossypetin, it was subsequently endocytosed much faster by mouse peritoneal macrophages. This modification did not occur at a concentration of 10 microM. Nine other flavonoids containing up to five hydroxyl substituents did not modify LDL to any great extent at 100 microM. The modification of LDL by 100 microM myricetin was time-dependent and complete by 6 hr. Flavonoids can sometimes act as pro-oxidants but myricetin did not act by oxidizing the LDL, as the LDL lipid hydroperoxide content was not increased by myricetin, nor did it promote the depletion of the endogenous antioxidant alpha-tocopherol in the LDL. High concentrations of myricetin caused the aggregation of LDL particles, as judged by light microscopy, agarose gel electrophoresis, retention by a membrane filter and sedimentability by centrifugation. SDS-PAGE indicated that the apolipoprotein B-100 molecules of LDL particles were covalently crosslinked. The uptake and degradation by macrophages of myricetin-modified 125I-labelled LDL reached saturation at about 10 micrograms protein/mL, suggesting the existence of a high affinity uptake process for the modified LDL. The uptake of myricetin modified 125I-labelled LDL was not competed for by a large excess of non-labelled native LDL or acetylated LDL. We conclude that myricetin and gossypetin at high concentrations are capable of modifying LDL by a novel non-oxidative mechanism to a form taken up by macrophages by a high affinity process. PMID- 8424825 TI - A role for Na/Ca exchange in the pancreatic B cell. Studies with thapsigargin and caffeine. AB - Sodium/calcium (Na/Ca) exchange is thought to play a role in Ca2+ extrusion from the pancreatic B cell. The aim of the present study was to provide direct evidence for such a role. The effect of extracellular Na+ (Na0+) removal on cytosolic free Ca2+ ([Ca2+]i) in single pancreatic B cells was examined using fura 2 and dual wavelength microfluorimetry. Isosmotical replacement of Na0+ by sucrose increased [Ca2+]i in the presence of extracellular Ca2+ but failed to affect [Ca2+]i in the absence of the divalent cation. Thapsigargin (1 microM), an inhibitor of the endoplasmic reticulum Ca(2+)-ATPase, induced a transient increase in [Ca2+]i in the presence of Na0+. This increase was enhanced and more sustained in the absence of Na0+. In the absence of Na0+ and the presence of thapsigargin, reintroduction of Na0+ induced a rapid decrease in [Ca2+]i. A similar picture was observed when caffeine (10 mM) was used to release Ca2+ from the endoplasmic reticulum. The decrease in [Ca2+]i induced by Na0+ reintroduction was accompanied by an important increase in 45Ca outflow from perifused islets. In conclusion, this study provides direct evidence that Na/Ca exchange may regulate B cell [Ca2+]i within physiological range. PMID- 8424826 TI - The role of intracellular free calcium mobilization in the mechanism of action of antitumour ether lipids SRI 62-834 and ET18-OMe. AB - Membrane-active antitumour ether lipids such as ET18-OMe (1-O-octadecyl-2-O methyl-rac-glycero-3-phosphocholine) and SRI 62-834 ((+-)-2-(Hydroxy[tetrahydro-2 (octadecyloxy) methylfuran-2-yl] methoxyl phosphinyloxy)-N,N,N trimethylethaniminium hydroxide) are selectively cytotoxic to tumour cells in vitro. Their precise mechanisms of action are unclear, but they are known to have effects on cell membranes and cell signalling. A previous report suggested that ether lipids cause a biphasic sustained rise in intracellular free calcium [Lazenby et al., Cancer Res 50: 3327-3330, 1990]. We show here that the second phase is an experimental artefact due to cell membrane permeabilization by ether lipids in serum-free buffers. In serum-free medium, the membrane toxicity of antitumour ether lipids was increased 50-60 fold, when compared to medium containing 10% serum. Membrane disruption was neither dependent on extracellular calcium, nor modulated by preloading cells with the calcium chelators bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetic acid or 2-[[2-[bis(carboxymethyl)amino] 5-methylphenoxy]methyl]-6- methoxy-8-[bis(carboxymethyl)amino]quinoline. This indicates that the mechanism of membrane damage by ether lipids does not involve changes in calcium homeostasis. Using indo-1 and fura-2 as calcium probes, we established that lower concentrations of antitumour ether lipids do elicit a genuine monophasic and transient rise in intracellular free calcium, predominantly mobilized from internal stores. This acute calcium agonist activity of ether lipids is distinct from the inhibitory effects on cell signalling reported previously after more prolonged exposure. It appears that the calcium elevation induced by antitumour ether lipids is unlikely to be instrumental in their selective and potent antitumour activity. PMID- 8424827 TI - Activity of cholinesterases in the Japanese quail embryo. Effects of dichlorphos on the embryonic development. AB - Cholinesterase activity is detectable in the Japanese quail embryo, in the yolk and subembryonic liquid, but not in the albumen. Obviously, this enzyme is deposited by the hen into the yolk and from there it is transferred to the subembryonic liquid. In contrast, in the embryo the enzyme is synthesized by itself and the amount increases with the age of the embryo. By using BW284c51 1,5 bis-(4-allyldimethylammoniumphenyl)pentan-3-one bromide and ISO-OMPA tetraisoprophylpyrophosphoramide as inhibitors, it was found that the enzyme in the embryo is predominantly acetylcholinesterase (EC 3.1.1.7), whereas that in the yolk and subembryonic liquid is butyrylcholinesterase (EC 3.1.1.8). Both types are inhibited by dichlorphos. However, the embryonic enzyme activity is restored within 8 hr, whereas that in the subembryonic liquid remained inactive at least for 72 hr after inhibition. Enzyme inhibition leads to retardation of the development, to reduced accumulation of glucose and amino acids in the subembryonic liquid and finally to death of the embryo, suggesting that the developmental retardation is due to the restricted supply of glucose and amino acids. Surprisingly, most of the embryos die when the embryonic enzyme activity has again been restored. PMID- 8424828 TI - 1992 Joseph J. Bunim Lecture. Pathogenetic concepts and origins of rheumatoid factor in rheumatoid arthritis. PMID- 8424829 TI - Suppression of adjuvant-induced arthritis in DA rats by incomplete Freund's adjuvant. AB - OBJECTIVE: To define the effects of incomplete Freund's adjuvant (ICFA) on subsequent arthritis induced by complete Freund's adjuvant (CFA) and type II collagen (CII) in DA and Lewis rats. METHODS: ICFA was injected into DA and Lewis rats before CFA or CII injection. RESULTS: DA rats previously injected with ICFA had significantly less severe arthritis induced by CFA compared with those not receiving ICFA pretreatment (P < 0.01). ICFA had no significant impact on CFA induced arthritis in Lewis rats or on CII-induced arthritis in DA rats. CONCLUSION: The injection of ICFA alone specifically suppresses subsequent CFA induced arthritis in DA rats, but not in Lewis rats. PMID- 8424830 TI - Nonperipheral accelerated nodulosis in a methotrexate-treated rheumatoid arthritis patient. PMID- 8424831 TI - Sjogren's syndrome and cutaneous B cell lymphoma revealed by anetoderma. PMID- 8424832 TI - Intravascular malignant lymphoma mimicking central nervous system lupus. PMID- 8424833 TI - HLA-DR4 subtypes in New Zealand Polynesians. Predominance of Dw13 in the healthy population and association of Dw15 with rheumatoid arthritis. AB - OBJECTIVE: To analyze HLA-DR4 alleles in New Zealand Polynesians with rheumatoid arthritis (RA). METHODS: Thirty Polynesians and 30 Caucasians with RA, as well as 65 Polynesian and 60 Caucasian healthy blood donors, were DR4 subtyped using the polymerase chain reaction and sequence-specific oligonucleotide probes. RESULTS: The frequency of DR4 (DRB1*04) was increased in both Polynesian (P < 0.001) and Caucasian (P < 0.005) RA patients compared with race-matched controls. Dw4 (DRB1*0401) was detected in 15 of 30 Caucasian patients but only 2 of 30 Polynesian patients (P < 0.001). In Polynesians, RA was associated with Dw15 (DRB1*0405), which was present in 11 of 30 patients and 3 of 65 controls (P < 0.001). Dw13 (DRB1*0403) was the most frequent DR4 allele in healthy Polynesians, but was not significantly associated with RA. CONCLUSION: The predominance of the Dw13 subtype in Polynesians may explain in part the low prevalence of RA in this population. The association of Dw15 with RA in Polynesians supports the hypothesis that the third hypervariable region of DR beta determines susceptibility to RA. PMID- 8424834 TI - Production of monocyte chemotactic protein-1 in human type B synoviocytes. Synergistic effect of tumor necrosis factor alpha and interferon-gamma. AB - OBJECTIVE: Since local secretion of chemotactic factors could contribute substantially to the homing of monocytes to the rheumatoid synovium, we investigated the ability of type B, or "fibroblast-like," synoviocytes isolated from the synovial tissue of patients with rheumatoid arthritis to synthesize and secrete the novel cytokine monocyte chemotactic protein 1 (MCP-1). METHODS: Synthesis and secretion of MCP-1 was determined by immunoprecipitation following metabolic labeling of MCP-1 with 35S-cysteine. MCP-1 gene regulation was assessed by Northern blot analysis. RESULTS: Unstimulated type B synoviocytes released little or no MCP-1, although low levels of MCP-1 messenger RNA (mRNA) were detected. However, incubation of these cells with tumor necrosis factor alpha (TNF alpha) resulted in a time- and dose-dependent release of MCP-1 into the supernatant, and expression of MCP-1 mRNA. Use of cycloheximide and actinomycin D confirmed that TNF alpha was inducing MCP-1 expression at both the transcriptional and translational levels. Treatment of the synoviocytes with interferon-gamma (IFN gamma) also stimulated an increase in both the steady-state levels of MCP-1 mRNA, as well as MCP-1 protein synthesis and secretion. In addition, TNF alpha and IFN gamma in combination exerted a synergistic effect on both MCP-1 mRNA accumulation and protein secretion. CONCLUSION: These studies demonstrate that the MCP-1 gene is regulated by TNF alpha and IFN gamma in type B synoviocytes and indicate that these cells may play an important role in the recruitment of inflammatory cells to the rheumatoid synovial environment, via the production of novel chemotactic cytokines such as MCP-1. PMID- 8424835 TI - Transforming growth factor beta exerts opposite effects from interleukin-1 beta on cultured rabbit articular chondrocytes through reduction of interleukin-1 receptor expression. AB - OBJECTIVE: The aim of the study was to determine whether transforming growth factor beta (TGF beta) modulates the effects and the receptor expression of interleukin-1 (IL-1) in rabbit articular chondrocytes (RAC). METHODS: Collagen, glycosaminoglycan, and collagenase production, together with 125I-labeled IL-1 binding, were analyzed in RAC cultures. RESULTS: TGF beta reduced both IL-1 effects on matrix metabolism and IL-1 receptor expression. CONCLUSION: TGF beta acts as an antagonist of the effects of IL-1 through down-regulation of its receptor expression. PMID- 8424836 TI - Degradation of human articular cartilage by neutrophils in synovial fluid. AB - OBJECTIVE: To determine whether surface-adherent immunoglobulins are capable of mediating synovial fluid (SF) neutrophil degradation of proteoglycan and collagen in intact, normal human articular cartilage, and to define the respective roles of neutrophil serine proteases and metalloproteases in degrading these cartilage constituents. METHODS: Pellet explants of normal human articular cartilage pretreated with bovine serum albumin (BSA) or IgG were incubated with polymorphonuclear cells suspended in SF (PMN-SF), or with supernatants derived from neutrophils stimulated with surface-associated IgG. Proteoglycan degradation was measured by assaying release of 35S-proteoglycan fragments from cartilage explants prelabeled with 35S-sulfate. Collagen degradation was measured by assaying hydroxyproline content in the PMN-SF preparations or neutrophil supernatants following their incubation with unlabeled explants. RESULTS: Significant release of both 35S fragments and hydroxyproline was noted following incubation of PMN-SF with IgG-treated pellets, compared with pellets treated with BSA. IgG preparations derived from pooled normal serum or rheumatoid arthritis SF were equally efficacious in mediating PMN degradation of cartilage collagens. Explant release of 35S fragments during incubation with PMN supernatant was completely inhibited when serine proteases were inactivated by diisopropyl fluorophosphate (DFP); however, release of 35S fragments was enhanced when metalloprotease activity was present in the supernatant. Release of hydroxyproline during incubation of explants with PMN supernatant was comparable in the presence of DFP or EDTA, but was markedly enhanced when both serine and metalloprotease activity were present in the supernatant. CONCLUSION: Neutrophils in SF are capable of degrading both proteoglycans and collagens in intact human articular cartilage. Degradation of these cartilage constituents is facilitated by immunoglobulins adherent to the cartilage surface and by the synergistic action of PMN serine and metalloproteases released during activation of neutrophils with surface-associated immunoglobulin. PMID- 8424837 TI - First international conference on rheumatic diseases in pregnancy. PMID- 8424838 TI - Therapy for childhood rheumatic diseases. Have we been doing enough? PMID- 8424839 TI - Fecundity before disease onset in women with rheumatoid arthritis. AB - OBJECTIVE: To examine fecundity prior to disease onset in women with rheumatoid arthritis (RA). METHODS: Two hundred fifty-nine RA patients with an opportunity for pregnancy were compared with 1,258 control women as part of a prospective case-control study of recent-onset RA in women. RESULTS: A 12-month or longer interval to pregnancy was found in 42% of RA cases compared with 34% of controls (odds ratio = 1.44, 95% confidence interval 1.10, 1.91). This difference was not accounted for by an older age when attempting to become pregnant among the RA cases compared with the controls. The result was unrelated to rheumatoid factor or HLA-DR4 status. CONCLUSION: Women with RA were found to have decreased fecundity prior to disease onset, compared with control women. PMID- 8424840 TI - Aggressive therapy for childhood rheumatic diseases. When are immunosuppressives appropriate? PMID- 8424841 TI - Cyclosporine in systemic sclerosis. Results of a forty-eight-week open safety study in ten patients. AB - OBJECTIVE: To evaluate safety and efficacy of cyclosporin A (CSA) treatment in systemic sclerosis (SSc). METHODS: Ten patients with < or = 60 months of SSc were entered into a 48-week open study of CSA. Patients with hypertension or azotemia were excluded. Concurrent use of nonsteroidal antiinflammatory agents or diuretics was not permitted. The extent of cutaneous and visceral involvement at 48 weeks and at study entry were compared. RESULTS: Adverse reactions (especially nephrotoxicity) were frequent, usually transient, often dose-limiting, and usually associated with CSA doses > or = 3-4 mg/kg/day. Skin thickening decreased significantly (P < 0.001), while pulmonary and cardiac involvement remained unchanged. CONCLUSION: CSA should undergo blinded controlled study in patients with SSc. PMID- 8424842 TI - Variation in the risk of peptic ulcer complications with nonsteroidal antiinflammatory drug therapy. AB - OBJECTIVE: To assess the risk of perforation or hemorrhage of peptic ulcer on treatment with nonsteroidal antiinflammatory drugs (NSAIDs), both as a class and as individual agents. METHODS: A case-control study of medication histories in 494 patients and 972 matched control subjects. RESULTS: The increase in risk (odds ratio) with NSAID therapy was 5.1 times the risk in controls. The odds ratio for piroxicam was 6.3 (95% confidence interval [CI] 3.3-12.0), as compared with 2.9 for diclofenac, ketoprofen, and sulindac combined (95% CI 2.0-4.2). The effect of other risk factors was also considered, and the adjusted odds ratios were 4.1 for all NSAIDs, 6.4 (95% CI 2.8-15.0) for piroxicam, and 3.3 (95% CI 2.0 5.5) for diclofenac, ketoprofen, and sulindac combined. CONCLUSION: The estimate of overall risk of peptic ulcer complications with NSAIDs is similar to that found in other studies. There appear to be differences in risk between agents. PMID- 8424843 TI - Cutaneous ulcers in a patient with systemic lupus erythematosus. PMID- 8424844 TI - A proposal for a model psychopharmacology curriculum for psychiatric residents. PMID- 8424845 TI - Rapid induction of conditioned opiate withdrawal in the rat. AB - Previous studies of conditioned opiate withdrawal in animals either have suffered from a lack of readily quantifiable data (e.g., measurement of diarrhea and vocalization in rodents) or were very long and costly (e.g., disruption of operant responding in monkeys). In this study, an attempt was made to produce a rapid and quantifiable measure of conditioned opiate withdrawal in the rat. Rats were trained to lever-press for food reinforcement on a fixed-ratio-15 schedule. All rats were then implanted with two subcutaneous 75-mg morphine pellets and allocated into three groups. The paired group received four naloxone injections (0.025 mg/kg SC) in the operant chambers paired with a distinctive tone and smell. The unpaired group was also exposed to the tone and smell in the chambers on four occasions, but received the naloxone injections in the home cage. The saline control animals were never exposed to naloxone or the tone and smell. On the test day, all rats were exposed to the tone and smell and injected with saline. The paired group showed a significant reduction in operant responding in response to the tone and smell when compared either with the other two groups, or to their own response rates on the previous day. In a second experiment, the paired and unpaired groups were again challenged with the tone and smell and a saline injection 1 month after removal of the morphine pellets. Again, the paired group showed a significant disruption of response. These results suggest that the conditioned stimulus acquired significant behavior-disruptive properties manifest even in the absence of opiate receptor occupancy.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424846 TI - LY248686, a new inhibitor of serotonin and norepinephrine uptake. AB - LY248686 is an inhibitor of serotonin (5-hydroxytryptamine; 5-HT) and norepinephrine (NE) uptake in synaptosomal preparations of hypothalamus and cerebral cortex, and 5-HT uptake in human blood platelets, with inhibitor constants near nanomolar concentrations. Upon administration to rats 1 hour before sacrifice, LY248686 caused dose-dependent and parallel decreases of 5-HT and NE uptake in hypothalamus homogenates ex vivo. LY248686 is a positive enantiomer and was slightly more potent than its negative isomer, LY248685, as an inhibitor of 5-HT uptake. Both isomers were only weak inhibitors of dopamine (DA) uptake in striatal synaptosomes. The inhibitory effects on 5-HT and NE uptake after a single administration of LY248686 followed similar time courses and simultaneously persisted for as long as 6 hours. LY248686 in vivo could effectively antagonize the p-chloroamphetamine-induced decreases of 5-HT uptake and levels of 5-HT and 5-hydroxyindoleacetic acid in cerebral cortex, and block the accumulation of 14C-NE in rat hearts. In food deprived rats, LY248686 suppressed food intake synergistically with 5-hydroxytryptophan, a precursor amino acid of 5-HT. Because of its lack of affinity for receptors of 5-HT, NE, DA, acetylcholine, histamine and naloxone, and its ability to inhibit 5-HT and NE uptake simultaneously, LY248686 has a favorable pharmacological profile as a potential antidepressant drug. PMID- 8424847 TI - Genetic differences in social behavior: relation to natural killer cell function and susceptibility to tumor development. AB - The hypothesis that certain heritable personality traits would correlate with increased vulnerability to tumor development and reduced natural killer (NK) cell function was tested in mice selectively bred for high and low levels of aggression. This selection program produces a line of mice that fail to exhibit species typical, isolation-induced aggression, but appear socially inhibited in response to a novel partner mouse. All socially inhibited mice developed 3 methylcholanthrene-induced tumors compared with only 44% of the aggressive mice. Basal NK activity was also significantly lower among socially inhibited mice. Conversely, there were no line differences in NK activity between the aggressive line and nonselected, socially isolated mice, consistent with other unidirectional outcomes of this selective breeding program. No significant line differences were present for nonsocial measures of emotional reactivity (e.g., fearfulness) or serum corticosterone levels. These findings support the hypothesis that social "traits" may be related to immune function and tumor susceptibility. PMID- 8424848 TI - Dose-related suppression of REM sleep and PGO waves by the serotonin-1 agonist eltoprazine. AB - Parental administration of the serotonin-1 agonist eltoprazine (0.0625 to 4.0 mg/kg [0.0002 to 0.016 mmol/kg]) in freely moving cats produced significant suppression of electrophysiologic rapid eye movement (REM) sleep signs, ponto geniculo-occipital (PGO) activity, and REM sleep behavior. The virtual total suppression of REM sleep (0.4%, 4.0 mg/kg) and PGO wave activity (2 to 4 mg/kg) in exchange for increasing amounts of non-REM (NREM) slow-wave sleep was a dose dependent function of the amount of eltoprazine administered. Wakefulness was unaffected by eltoprazine regardless of dose. Concurrent with this dose-dependent suppression of REM was a dose-dependent increase in electroencephalographic synchrony and mean electromyographic amplitude. Since eltoprazine was found to shift the balance between REM and NREM sleep but did not change the balance between sleep and waking, it is a potentially useful tool for the investigation of serotonergic-cholinergic interaction. PMID- 8424849 TI - Effects of the novel acetylcholinesterase inhibitor SDZ ENA 713 on sleep in man. AB - A novel brain-selective acetylcholinesterase inhibitor, SDZ ENA 713, is under development for the treatment of dementia of the Alzheimer type. To determine the threshold dose for central activity, single doses of the compound were administered to 20 young male volunteers in a double-blind cross-over design and the effects on the sleep electroencephalography studied. The first group of eight volunteers received in random order: placebo, 0.5 mg; and 1 mg SDZ ENA 713. The second group of 12 volunteers received: placebo, 1.3 mg; and 2 mg SDZ ENA 713. Sleep quality was not affected by the study medication, which was well tolerated by all subjects. A statistically significant increase in rapid-eye movement sleep density was observed after doses of 1 mg, 1.3 mg, and 2 mg. Rapid-eye movement latency and slow-wave sleep were not altered. The results demonstrate that SDZ ENA 713 is centrally active in man at well-tolerated doses. PMID- 8424850 TI - Perception of taste and smell in elderly persons. AB - By the turn of the century, there will be considerable shifts in demographics, including a massive increase in our aging population. As we plan for better nutrition in the twenty-first century, the special sensory and nutritional needs of the elderly must be taken into account. Chemosensory losses, specifically decrements in the senses of taste and smell, can lead to inadequate intake, especially in the elderly sick. These losses result not only from anatomic changes that occur during normal aging but also from certain diseases, pharmacological and surgical interventions, radiation, and environmental pollutants. The design of foods for the elderly that could both compensate for these chemosensory losses and meet nutritional needs presents new challenges and opportunities for the food industry. PMID- 8424851 TI - Changes in taste and flavor in aging. AB - This study aims at understanding the role of mixtures (mutual quality suppression) in the evaluation of impact of the human aging process on the perception of taste. Heretofore, the effect of aging on taste has been directed at threshold and suprathreshold magnitudes of single chemicals (e.g., NaCl, sucrose, citric acid) in aqueous solution. Although absolute thresholds typically rise in advanced age (2 to 9 times, depending on the study), suprathreshold magnitude assessed by magnitude matching seems (except for bitter) to resist change in the way presbycusis spares suprathreshold loudness, fostering the impression that aging may handicap the aged little in the perception of food. Asked, however, to discriminate the presence-absence of the prescribed salt flavoring (nominally suprathreshold) in tomato soup, the young outperformed the middle-aged who, in turn, outperformed the elderly. Moreover, NaCl thresholds in the presence of tomato measured several times higher than in water, but the difference between the young and the elderly continued to hold. Elevation of threshold to much higher levels by mixture suppression leaves the young-elderly difference unchanged, implying that the elderly may fail to detect salt levels that really count in their diet. To examine the relation between age and taste mixtures, we measured detection thresholds: (1) for NaCl in citric acid, from zero to strong; (2) for sucrose in citric acid, from zero to strong; and (3) for citric acid in sucrose, from zero to strong. Whether in water alone or in a weak or strong suppressor, the elderly subjects' threshold was consistently 2 or 3 times higher than that of the young. Moreover, the way in which threshold for one quality rises with concentration of a suppressor is the same, except for constant upward displacement of the elderly peoples' threshold. In general, both young and elderly confuse salty and sour (show large suppression at all concentrations of the suppressor) much more than they confuse sweet and sour (seen mainly at high concentrations of the suppressor). Study of other mixtures is planned. PMID- 8424852 TI - Nutrition and chemosensory perception in the elderly. AB - The elderly person's perception of foods and food flavor is affected by age associated changes in the chemical senses (taste, smell, and trigeminal sensation). Both classic and modern psychophysical techniques have been applied to achieve some understanding of these changes. Taste threshold sensitivity declines with age; however, the magnitude of the decline and the degree to which taste qualities are differentially affected remains to be understood. Suprathreshold taste intensity perception is affected by age, but there is a differential effect of taste quality. Experiments with blended foods have indicated that both olfactory and taste deficits contribute to older people's difficulty with food identification. Experiments assessing threshold sensitivity, suprathreshold intensity, and suprathreshold identification have all demonstrated significant impairment in olfaction in old age. In fact, these effects are far greater than in the taste system. Patients with Alzheimer's disease show even greater olfactory deficits than normal elderly and the effect is related to the degree of dementia. We have ruled out nasal disease in these patients as the primary causes of olfactory insensitivity, because significant impairment remains when the influence of nasal airflow and nasal cytology has been removed statistically. Both normal elderly and patients with Alzheimer's disease show impairment in odor memory. Sensitivity to, familiarity with, and identifiability of the odors all play a role in odor memory performance. Flavor preference also changes over the lifespan. Older subjects, for instance, rate high concentration of sugar and salt as pleasanter than young subjects do. Multiple influences contribute to this phenomenon. Elderly persons and those of lower nutritional status have shown preferences for higher concentrations of casein hydrolysate than young persons and those of higher nutritional status, suggesting that dietary preference can be related to chemosensory cues. There are significant age associated changes in chemosensory perception that have the potential to interact with dietary selection and nutrition in the elderly. A better understanding of these phenomena may promote health and well-being in the elderly. PMID- 8424853 TI - Appetite, hunger, and satiety in the elderly. AB - Food intake declines with age and the elderly frequently report a decrease in appetite. Social and physical factors as well as decreased energy expenditure may be part of the explanation for the decreased food intake; however, it is possible that changes in hunger mechanisms are also involved, but more studies similar to those conducted on thirst are needed. The consumption of a varied diet depends in part on sensory-specific satiety, that is, the decrease in the pleasantness of a food as it is consumed. We have shown that sensory-specific satiety changes with age such that it was absent in individuals over the age of 65 and this could be part of the explanation of why the elderly consume more monotonous diets. Additional basic studies on how aging affects the controls of food intake will suggest ways to improve the nutritional status of the elderly. PMID- 8424854 TI - Tailoring texture for the elderly: theoretical aspects and technological options. AB - The perception of what is known as "texture" is a result of a variety of primarily mechanical and acoustical events. These are sensed by receptors of different kinds that are unevenly distributed in the sensing organ(s) (tongue, jaws, fingers) and neighboring tissues. The perceived intensity of textural properties is time dependent and can be influenced by other stimuli, notably chemical and thermal. The exact mode in which mechanical and geometrical properties are translated into perceived texture is not known. There are, however, known trends based on empirical observations, and growing understanding of the operation of individual receptors and the physiology and biomechanics of the sensation process itself. This body of knowledge can provide rational guidelines for the development of special products for the elderly and other people with special needs. For example, it is possible to increase fragility and maintain crunchiness or to make chewy foods that require a reduced effort in mastication to minimize fatigue. The technology to make such products is already in place and its future use by the food industry will depend only on marketing opportunities. PMID- 8424855 TI - Psychophysiological effect of odor. AB - Techniques to measure the activity of the taste and odor molecules at the olfactory epithelium, taste bud, and brain response levels are now being used to determine and measure the actual transduction of the chemical information to the brain waves. We studied a number of methods of measuring brain wave responses to odorants and we settled on an electroencephalographic method of measuring the slow brain waves [> 13 Hz] at the frontal location on the scalp. This technique known as contingent negative variation (CNV) measures the early component [at 400 to 1000 ms] of the beta wave variation, which can be negative, positive, or neutral depending on the odorant being presented to the subject. This component is almost independent of the subject's psychological state, degree of arousal, or level of consciousness and is known as the external component. The experimental paradigm creates a reproducible result in which odorants may be classified as stimulating, sedating, or neutral. These psychophysiological effects of odors on man appear to offer a means to determine precognitive responses directly related to the effect of the chemical messenger. There does not appear to be any bias as to sex, national origin, or race. At this point in our research efforts, we do not see evidence that there is a bias related to age. It is proposed that some of the problems of classical sensory evaluation can be helped by the use of psychophysiological recording techniques, such as CNV, as a measurement of brain activity and response to flavor and aroma. PMID- 8424856 TI - The effects of aging on the human sense of smell and its relationship to food choice. AB - Olfaction plays a significant role in the perception of foods. For the most part, taste is limited to sweet, sour, bitter, and salty. The sensory experiences during consumption of complex foods and drinks cannot be constructed from these units. Indeed, much of the taste of a meal derives from olfactory stimulation. Hence, factors that influence olfactory perception should affect treatment of food-related odors. This article initially reviews previously published observations on the effects of age on olfaction and food preferences and then presents the results of original analyses of data derived from a substantial database formed as a result of the National Geographic Smell Survey. Included in the Survey form were topics relevant to the present article. They include the following question: Would you eat something that smelled like this? Two of the odors in the Survey were food related and two were fragrance related. Hence, in addition, we assessed responses to the following question: Would you apply something that smelled like this to your body? Answers were affected in part by the age and gender of the respondent and by the perceived pleasantness and intensity of the odor. PMID- 8424857 TI - Color as a factor in food choice. AB - From birth, nature teaches us to make judgements on our environment based in large measure on color. As such, it plays a key role in food choice by influencing taste thresholds, sweetness perception, food preference, pleasantness, and acceptability. Its role is elusive and difficult to quantify, however, which at times has placed color in a secondary role to the other sensory characteristics, a position not entirely consistent with the facts. Color, in a quantitative sense, has been shown to be able to replace sugar and still maintain sweetness perception in flavored foods. It interferes with judgments of flavor intensity and identification and in so doing has been shown to dramatically influence the pleasantness and acceptability of foods. Studies in the literature have used cross-sectional population panels to study these effects, but a recent investigation of color-sensory interactions in beverages has compared the response of a college age group with the response of a panel consisting of a more mature population. Interestingly, the older group showed significant differences from the college age group in their response to the effects of color on several sensory parameters as well as showing a direct correlation between beverage consumption and color. Color is often taken for granted, but this position must be reevaluated in view of such studies and the need to create more appealing foods for different segments of our society. PMID- 8424858 TI - Neuropsychological sequelae of Wernicke's encephalopathy in a 20-year-old woman: selective impairment of a frontal memory system. AB - This article reports the neuropsychological sequelae of Wernicke's Encephalopathy in a 20-year-old woman. After amelioration of acute symptoms, the patient showed a range of cognitive impairments the most marked of which was a severe impairment on free recall tasks involving both verbal and nonverbal material. In contrast, recognition memory appeared remarkably well preserved. Some evidence of frontal lobe and general intellectual impairment was also noted. The case is discussed in terms of the pathophysiology of Wernicke-Korsakoff Syndrome, criteria for its diagnosis, and the multicomponent nature of memory deficits. PMID- 8424859 TI - Perception of human chimeric faces by chimpanzees: evidence for a right hemisphere advantage. AB - The aim of this study was to investigate hemispheric advantages in the processing of human chimeric faces by chimpanzees using a free-vision task developed by Levy, Heller, Banich, and Burton (1983a). Subjects were taught a visual discrimination in which they were to select the human face that appeared "happiest" when paired with its neutral counterpart. After reaching criterion, chimeric faces were substituted as test trials, and the face the subjects selected was recorded. On 62% of the trials, three chimpanzee subjects selected the face in which the smiling half fell in the left hemispatial field. Familiarity factors appear to have accounted for some of the observed findings. We believe this is the first evidence for hemispheric advantages in the perception of human faces by great apes. PMID- 8424860 TI - On the complex relation between perceptual characteristics and hemispheric asymmetry. AB - The relation between hemispheric asymmetry and perceptual characteristics in general, and the spatial frequency content of input in particular, are discussed in reference to Peterzell's (1991) reply to a literature review by Christman (1989a). Three main points are presented: (1) there is evidence that spatial frequency, independent of the total information/energy content of input, does influence patterns of hemispheric asymmetry; (2) that information and energy represent potentially dissociable constructs that need to be considered separately; and (3) that there is recent evidence that numerous input and task characteristics, other than spatial frequency content, influence hemispheric asymmetry (e.g., spatial phase, relative spatial frequency, temporal frequency, task requirements, etc.). PMID- 8424861 TI - Memory for source after traumatic brain injury. AB - A fame judgment task was used to distinguish subjects' ability to recognize previously presented information from their ability to recognize the source of that information. Traumatic brain injured (TBI) subjects were impaired relative to controls with respect to verbal recall and recognition as well as memory for source. However, memory for source was demonstrated to be independent of explicit indices of recall and recognition ability. It was also found to be an extremely sensitive index of coma duration in the TBI subjects. The anticipated relationship between source memory and a putative index of frontal function (Wisconsin Card Sorting Test) was modest relative to the relationship between source memory and subjects' performance on a complex visual pattern matching task (Benton Facial Recognition), raising questions about hypothesized cognitive and neurophysiological mechanisms underlying memory for source. PMID- 8424862 TI - Subcortical contributions to drawing. AB - Constructional impairment is often considered a sign of cortical damage. However, aphasia, agraphia, and apraxia, disorders traditionally deemed cortical, have been well described following subcortical lesions, suggesting an important role for subcortical structures in cognition. Drawing impairment following subcortical lesions has not been systematically explored or compared with that following cortical damage. We examined relative incidence and severity of drawing impairment after cortical and subcortical strokes and whether there were qualitative differences between these two groups' drawings. Drawings by 125 patients with single hemispheric strokes of similar volume (42 left cortical, 36 left subcortical, 20 right cortical, 27 right subcortical) were compared using a standardized scoring system. Although previously noted right/left differences were confirmed, "subcortical drawings" did not differ from "cortical drawings" on any measures, including overall impairment. Compared with cortical patients, drawing impairment in those with subcortical lesions (especially left) was more strongly associated with impairment of other cognitive abilities. Thus, although a subcortical lesion does not cause more severe drawing impairment, subcortical lesions affecting drawing lead to more widespread cognitive dysfunction than do similarly sized cortical lesions. Drawing impairment often follows subcortical strokes and is by no means an indicator of cortical lesion localization. PMID- 8424863 TI - Sex differences in mental rotations: an effect of time limits? AB - It has been argued that the male performance advantage on the Mental Rotations Test, a measure of three-dimensional rotational ability, reflects a slower problem-solving strategy in females. A recent study of a high-ability sample suggested that sex differences do not reach significance when the test is administered without time limits (Goldstein, Haldane, & Mitchell, 1990). To further explore the effects of time limits, unspeeded versions of the Mental Rotations Test and a vocabulary test were administered to 88 males and 94 females, who represented a less restricted range of ability levels. Males obtained significantly higher scores than females on the Mental Rotations Test (p = .0001), despite comparable performance for the two groups on the vocabulary test. In addition, distributions of Mental Rotations scores were significantly different, with greater proportions of men at the high and women at the low ends. Thus, a difference in speed of problem solving strategy is not a sufficient explanation of the sex difference on the Mental Rotations Test. PMID- 8424864 TI - Hemispheric specialization for handwriting in right handers. AB - Hand preference may be related to either lateralized language, movement representations, or both. Anatomic and behavioral studies have revealed that whereas each hemisphere has motor control of the contralateral distal and proximal limb movements, this same hemisphere's control over the ipsilateral limb is limited to proximal movements. This differential proximal-versus-distal organization may have functional implications such that when right handers write with their left hand they must be use more proximal musculature than when they write with their right hand. If hand preference is related to lateralized movement representations, right handers may also use more proximal than distal movements when drawing with their left than with their right hand. If one uses distal musculature to write or draw, the elbow travels through space less than if one uses proximal musculature. We studied 12 right handers by having them write and draw with their right and left hands and measured the spatial amplitude of their elbow movements. We found that when writing or drawing, right handers moved their left elbow more than their right. These results suggest that it is the lateralized movement representations that may be primarily responsible for writing hand preference. PMID- 8424865 TI - Slowing of information processing in Parkinson's disease. AB - In this study, we criticize the notion of bradyphrenia and argue that "slowness of thought" in Parkinson's disease (PD) must be analyzed as slowness of different information-processing stages and that unselected patients should not be used in experimental studies. We selected 32 patients with a long history of PD and 50 control subjects. Sixteen patients had mild cognitive deterioration (not dementia) and 16 patients had preserved cognitive capacities; otherwise the groups were matched. By using computerized tests, we investigated three separate stages: automatic and controlled processing and motor programming. The results indicate that patients with mild cognitive deterioration are slower than patients with preserved cognitive capacities or controls in automatic visual and in controlled processing but not in motor programming. We conclude that the slowing of controlled processing reflects the disruption of central neural networks, that a long history of PD does not necessitate cognitive slowing, and that PD is not a neuropsychologically serviceable category. PMID- 8424866 TI - High-throughput PCR. AB - The number of PCR samples that can be simultaneously processed has been dramatically increased over existing practices by using a new polycarbonate 864 well microwell plate and a modified air cycling oven. In thirty 9-min cycles, four plates containing 3455 samples can be amplified in 4.5 h. Amplification is rapid, uniform and reliable from sample to sample and run to run. This PCR method can satisfy the Human Genome Project's need for high-throughput sample analysis using PCR. PMID- 8424867 TI - Automated production of high density cosmid and YAC colony filters using a robotic workstation. AB - We report here on a system for automated preparation of high-density colony filters of arrayed libraries using the high density replicating system (HDR) for the Beckman Biomek 1000 robotic workstation. This system, consisting of a 96-pin tool, a sterilization station and controlling software, transfers samples from microplates onto target membranes in arrays up to 36 times the density of a 96 well microplate. The transfer operation can be completely automated with the addition of the Biomek Side Loader System, which consists of a robotic arm capable of transferring plates and filters between the Biomek working tablet and a storage area. Using the complete system, we are able to plate 32 replica filters (8 x 12 cm), each containing the clones from 16 different microplates (i.e., 1536 clones per filter), in a 16-h overnight run without any operator intervention. We describe conditions used for transfer of bacterial yeast colonies and fixation of DNA to the membranes, and we illustrate hybridization results obtained with cosmid and YAC filters. PMID- 8424868 TI - Automated fluorescent primer extension. PMID- 8424869 TI - PCR used to determine mating type in S. pombe. PMID- 8424870 TI - A rapid method for generating linker scanning mutants utilizing PCR. PMID- 8424871 TI - Rapid and simple purification of PCR products by direct band elution during agarose gel electrophoresis. PMID- 8424872 TI - Automated "hot start" PCR using mineral oil and paraffin wax. PMID- 8424873 TI - Paraffin beads can replace mineral oil as an evaporation barrier in PCR. PMID- 8424874 TI - Use of oil overlays in "oil-free" PCR technology. PMID- 8424875 TI - Three-color immunofluorescence imaging of Drosophila embryos by laser scanning confocal microscopy. AB - We present a simple means for triple-labeling biological specimens by immunofluorescence using a laser scanning confocal microscope for imaging with a krypton/argon laser as a light source. Three separate images of fluorescein-, lissamine rhodamine- and cyanine-5-labeled antibodies are collected and subsequently merged to form the triple-labeled image, which is displayed at full image resolution (24 bit) on a second image processing system. The technique is illustrated using immunofluorescence localization of three segmentation proteins in Drosophila embryos. PMID- 8424876 TI - Monitoring fusion protein cleavage by capillary electrophoresis. AB - Capillary electrophoresis was successfully utilized to monitor the proteolytic cleavage of two fusion proteins; maltose binding protein-sugarcane mosaic potyvirus coat protein and maltose binding protein-paramyosin. The course of proteolysis by factor Xa was monitored with high sensitivity and virtually in real time. Capillary electrophoresis conditions were optimized to clearly resolve the fusion proteins, factor Xa, lysis products and buffer components. Rapid elucidation of time to complete digestion was possible and fate of released proteins could also be examined. PMID- 8424877 TI - Post-capillary Immobilon-P membrane fraction collection for capillary electrophoresis. AB - A simple, cost-effective and efficient method of continuous fraction collection for capillary electrophoresis is described. The method employs a membrane, such as polyvinylidene difluoride, onto which the separated components from a capillary electrophoresis capillary are directly collected to overcome the problem of analyte dilution and to facilitate the fraction collection process. Model proteins are separated and collected onto a polyvinylidene difluoride membrane. PMID- 8424878 TI - In vitro transcription of RNAs with defined 3' termini from PCR-generated templates. AB - We demonstrate the feasibility of using PCR to economically amplify sufficient template to permit the transcription by T7 RNA polymerase of preparative amounts of RNAs for biochemical analyses. We show that a standard 100-microliter PCR amplification of a fragment from the 3' end of the genomic cDNA of turnip yellow mosaic virus yields enough template to support the synthesis of about 50 micrograms of a 264-nucleotide-long transcript. The choice of the 3' primer defines the 3' terminus of the transcripts, although, as with transcription from DNA linearized by restriction digestion, a subpopulation of transcripts with one or two additional 3' nucleotides is present. This PCR-based approach can be adapted to the rapid generation of RNAs with different 3' termini and with mutations near the 3' end. PMID- 8424879 TI - Expression cloning exploiting PCR rescue of transfected genes. AB - Expression cloning in eukaryotic cells has allowed the isolation of many genes for which no protein or nucleic acid sequence information was available. In this paper we describe a model methodology for PCR-mediated rescue of transfected genes from eukaryotic cell lines after functional selection. This protocol should allow any readily transfectable cell line to serve as a host for functional selection of cDNA expression libraries. PMID- 8424880 TI - An easy, safe technique for removing caps from screw cap microcentrifuge tubes containing 32P. PMID- 8424881 TI - Quantitative competitive polymerase chain reaction for accurate quantitation of HIV DNA and RNA species. AB - Inherent features of the PCR make this procedure suboptimal for quantitative applications. For typical clinical specimens, the absolute amount of product derived from PCR does not always bear a consistent relationship to the amount of target sequence present at the start of the reaction. Competitive PCR approaches to quantitation of nucleic acid sequences overcome the limitations of basic PCR methods for quantitation. We have developed a competitive PCR method known as quantitative competitive PCR for quantitation of HIV DNA and RNA sequences. Key features of the technique include quantitation, based on the relative amounts of products produced from the target sequence and a competitive template introduced into test specimens, and stringent internal control of all reactions, including the reverse transcription step in RNA PCR. The method is suitable for analysis of clinical specimens and may be particularly valuable for accurate quantitation of viral load in patients undergoing treatment with experimental therapies. PMID- 8424882 TI - Use of an automated workstation to facilitate PCR amplification, loading agarose gels and sequencing of DNA templates. AB - A series of protocols were developed for a commercially available automated workstation to prepare samples for amplification of DNA by the polymerase chain reaction (PCR) on automated thermal cyclers, to load PCR products onto agarose electrophoretic gels and to carry out dideoxynucleotide sequencing of DNA templates. The protocols and the software programs developed reduced by two thirds the time required to carry out the procedures manually. The programs and protocols also improved the quality and the consistency of both the PCR products and the sequencing reactions. PMID- 8424884 TI - Viewing a mosaic: caring in nursing services. PMID- 8424885 TI - Defining nursing administration terms (continued). PMID- 8424883 TI - Capillary electrophoresis: a powerful microanalytical technique for biologically active molecules. AB - Capillary electrophoresis (CE) is a versatile microanalytical technique that has gained much attention, particularly from those working with biologically active molecules. Its appealing characteristics include unprecedented sensitivity and the ability for automating the rapid electrophoretic separation of a number of low-volume samples in a reproducible manner, with relatively short analysis times. The picomole-femtomole (10(-12)-10(-15) mol) sensitivity of UV-CE has been enhanced tremendously by the interfacing of detection systems such as laser induced fluorescence, which has extended the sensitivity into the attomole zeptomole (10(-18)-10(-21) mol) range. Fluorescence detection has shown great potential for the CE analysis of a wide range of biomolecules including peptides, proteins and DNA. CE research and development has taken on directions focused primarily on improving detection, understanding and exploiting the basic chemistry of CE and devising new applications. PMID- 8424887 TI - CDR and EPI technical briefings at WHO headquarters. PMID- 8424886 TI - Spectrophotometric determination of clodronate with thorium-morin. PMID- 8424889 TI - Novel oxidative cleavage of C17-C20 bond in pregnane by a Pseudomonas sp. AB - Oxidative cleavage of the C17-C20 bond in progesterone by Pseudomonas sp. is reported. Transformation occurred under in vivo conditions. The bioconverted products were characterized as 1,4-androstadien-3,17-dione and 17 beta-hydroxy 1,4-androstadien-3-one. The steroid nucleus was not further degraded although the test organism had the capacity to induce dehydrogenation at C1 of this alpha, beta-conjugated steroid. PMID- 8424888 TI - Expression and functional analysis of steroid receptor fragments secreted from Staphylococcus aureus. AB - Fragments of the DNA-binding domain of the rat glucocorticoid receptor (rGR) and the human estrogen receptor (hER) were expressed in Staphylococcus aureus as a chimeric fusion to protein A by using a modified expression vector with an artificial factor X-cleavage site. The secreted product was isolated by hydrophobic chromatography on Phenyl-Sepharose and purified on DNA-cellulose or by anion-exchange chromatography. After cleavage of the protein A moiety, the purified rGR DNA-binding domain from amino acids 406 to 523 (rGR406-523), binds specifically to a glucocorticoid responsive element as a homodimer but cannot form heterodimers with the DNA-binding domain of the hER. Amino acids 510 to 523 following the zinc finger region, as well as free sulfhydryl-groups are necessary for DNA-binding, which is more efficient when the tripeptide Gly-Gly-Cys is added to the carboxy terminal end. Despite its specific interaction with DNA, rGR406 523 does not activate transcription from the MMTV promoter in a cell-free system that efficiently responds to addition of native GR, suggesting that regions essential for transcriptional activation in vitro are located outside of the DNA binding domain. PMID- 8424890 TI - Testosterone regulates mitochondrial aspartate aminotransferase gene expression and mRNA stability in prostate. AB - The effect of testosterone on the precursor mitochondrial aspartate aminotransferase (pmAAT) gene and on pmAAT-mRNA was studied in rat ventral prostate (VP) and pig prostate epithelial cells. Castration significantly decreased the level of nuclear pmAAT transcripts in VP; whereas testosterone treatment of castrated animals restored the level of pmAAT transcripts. Correspondingly, castration resulted in a marked decrease in the transcription rate of the pmAAT gene; whereas testosterone treatment markedly increased the transcription rate. In vitro studies with isolated pig prostate epithelial cells demonstrated that testosterone directly and rapidly induced a transient increase in the transcription rate of the pmAAT gene. The increase in transcription was associated with an increase in the steady-state level of pmAAT-mRNA. Similar in vitro effects were observed with isolated VP epithelial cells. In addition to its stimulatory effect on transcription of the pmAAT gene, testosterone also increased the half-life of pmAAT-mRNA from 2 h in the absence of hormone to 16 h in its presence. Consequently, testosterone appears to stabilize the pmAAT-mRNA. The combination of its immediate effect on stimulating the transcription of the pmAAT gene and its stabilizing effect on pmAAT-mRNA would account for the increase in the steady-state level of pmAAT-mRNA by testosterone. These studies support our proposal that, through these effects, testosterone increases the biosynthesis of mAAT thereby increasing the transamination of aspartate to oxaloacetate and ultimately increasing the synthesis of citrate. This appears to provide at least one of the mechanisms by which testosterone regulates prostate citrate production. PMID- 8424891 TI - Kinetics and metabolism of 11-deoxycortisol in a patient with congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency. AB - The kinetic features of 11-deoxycortisol (S) were studied in a 11 beta hydroxylase deficient boy. After i.v. administration of 35 kBq [3H]S (11 pmol) together with 44 nmol [13C]cortisol all his urine was collected during the next 3 days. A recently reported kinetic model, by which the fate of radioactive cortisol (F) in the body can be described by analysis of only the urinary radioactivity, has been used to calculate the rate constants of S metabolism. The overall half-life of S in the circulation was 4.7 min, which is very close to a reported half-live of the rapid phase: 4.1 min determined from the plasma radioactivity. The time of maximal accumulation of S in the first metabolic pool- 26 min is about one quarter of that found for F--109 +/- 20 min (n = 8). The half live of the S metabolites in the body was 7.0 h, equal to that of F: 6.1 +/- 0.9 h (n = 8). Obviously S is taken up into the metabolic organs 4 times faster than F, but it is not metabolized faster. The production rates of S and F were 127 and 2.1 mumol/(m2*d), respectively, pointing to a severely deficient synthesis of F. However, from the urinary excretion of 3 alpha,21-dihydroxy-5 beta-pregnan-20-one in relation to 3 alpha,11 beta,21-trihydroxy-5 beta-pregnan-20-one it cannot be concluded that the synthesis of corticosterone was strongly impaired. PMID- 8424892 TI - Dimerization of the RU486-bound calf uterine progesterone receptor. AB - Dimerization of the RU486-bound progesterone receptor was studied by measuring the Hill coefficient of RU486 binding in calf uterine cytosol at receptor concentrations between 0.3 and 15 nM. The limiting value of the Hill coefficient at high receptor concentrations was 1.38 +/- 0.01. The limiting value of the Hill coefficient at low receptor concentrations was 1.05 +/- 0.03. The dimerization constant, defined as the concentration of receptor at which the Hill coefficient was midway between the limiting values, was 2.6 +/- 0.1 nM. In contrast, the dimerization constant of the progesterone-bound receptor, which was measured using the same approach, is 7 nM [Skafar, D. F. Biochemistry 30 (1991) 6148 6154]. The results presented here support and quantify the observation that the RU486-bound progesterone receptor will dimerize at lower receptor concentrations than the progesterone-bound receptor. PMID- 8424893 TI - Changes in population density elicit quantitative and qualitative changes in the estrogen receptor in intact GH4C1 pituitary tumor cells. AB - We have previously demonstrated that population density alters the responsiveness of GH4C1 pituitary tumor cells to 17 beta-estradiol (E2). At a low population density E2 was observed to increase prolactin mRNA and stimulate cell proliferation, whereas this estrogen was unable to elicit these responses when the cells were maintained at a 4-fold higher population density. In an attempt to determine the mechanism through which population density alters responsiveness to E2, the steady-state level of estrogen receptor (ER), the affinity of ER for E2, and ER down-regulation have been examined in both intact and fractionated cells using ligand binding and ligand exchange assays. Data presented herein demonstrate that (1) GH4C1 cells maintained at low density expressed fewer ER than cells cultured at high density; (2) ER in cells cultured at high density displayed a reduced affinity for E2; (3) ER down-regulation occurring within 1 h of E2 addition appeared to be more pronounced in high density cultures; and (4) steady-state levels of ER were similar in low and high density cells treated with E2 for 1 through 5 days. Although none of these observations appear to correlate with the previously observed effects of population density on the responsiveness of GH4C1 cells to E2, they further illustrate the potential of the culture environment to alter the responsiveness to estrogenic stimuli by altering the properties of the ER. PMID- 8424894 TI - The role of cytochrome P450 3A (CYP3A) isoform(s) in oxidative metabolism of testosterone and benzphetamine in human adult and fetal liver. AB - Testosterone metabolism was studied in human adult and fetal liver microsomes. In fetal livers 6 beta-hydroxylase (6 beta OH) activity (1-2% of adult activity) and 2 alpha-hydroxylase (2 alpha OH) activity (about 40% of adult activity) were present. Also some fetal livers produced two unknown metabolites. Androstenedione was formed in all fetal livers studied (10-20% of adult activity). Testosterone hydroxylations at 6 beta-, 2 beta-, 15 alpha- and 15 beta-positions were associated with CYP3A isoform(s) in adult liver, because they were strongly inhibited by midazolam, a known substrate for CYP3A4 and by anti-CYP3A4 antibody. Fetal liver activities were consistently inhibited less than the activities in adult livers. The formation of androstenedione was not affected by these inhibitors in fetal or adult liver microsomes. Benzphetamine N-demethylase activity in the fetal livers was about 40% of adult activity. Anti-CYP3A4 antibody had no effect on that activity in fetal or in adult liver microsomes, whereas a monoclonal antibody 1-68-11 (generated against rat CYP2C11) slightly inhibited benzphetamine N-demethylase activity in adult liver. This study indicates that human fetal and adult liver are dissimilar in their testosterone metabolism pattern. The formation of androstenedione from testosterone in fetal liver may have a physiological role. Testosterone hydroxylases are less inhibited by anti-CYP3A4 antibody, midazolam and progesterone in fetal than in adult liver. PMID- 8424895 TI - Metabolism of progesterone and testosterone in human parotid and submandibular salivary glands in vitro. AB - Twenty-three parotid and twelve submandibular human salivary glands were used for analyzing in vitro metabolism of progesterone (P) and/or testosterone (T), in order to find out their possible contribution to the concentrations of salivary P or T. Glands were excised from 21 women and 14 men because of a stone, tumor or sialoadenitis, and the healthy tissue was used for incubation. The tissues were homogenized and incubated with 5 to 7 nmol/l of [14C]P or [14C]T for 2 h, extracted with methylacetate and subjected to two dimensional thin-layer chromatography. The thin-layer plates were autoradiographed, and the radioactivity of the different metabolites were counted to assess the amounts of metabolites formed. No lipoidal or conjugated steroids were formed. All of the radioactivity was associated with the free fraction of steroids. Both P and T were metabolized significantly more actively in male submandibular gland compared to male parotid gland (P < 0.05-0.01). No significant differences were found between female and male parotid glands, nor between the parotid glands of hormonally medicated (oral contraceptives or postmenopausal estrogen treatment) and non-medicated women. The submandibular glands more actively metabolized the steroids studied than the parotids (P < 0.01). The main metabolites were 20 alpha hydroxy-4-pregnen-3-one for P, and androstenedione for T. In conclusion, the present results bring further evidence that the P or T levels in saliva may not be identical with the unbound steroid fraction in circulation, but a part of the steroids are metabolized in salivary glands. However, due to the rapid passage of steroids from blood to saliva, the metabolism demonstrated probably does not form an important source of error in salivary P and T measurements. PMID- 8424896 TI - Beta-adrenoceptor mediated responses and subtypes of beta-adrenoceptors in cultured rat Sertoli cells. AB - Membrane particles from Sertoli cell cultures were examined for subtypes of beta adrenoceptors with a radioligand binding technique using [125I]iodocyanopindolol and a beta 1-selective antagonist (Sandoz 204 545) or a beta 2-selective antagonist (ICI 118 551). Biphasic competition curves and modified Eddie-Hofstee plots revealed a relative distribution of approx. 80% beta 1-adrenoceptors and 20% beta 2-adrenoceptors. Only 45% of the adrenoceptor mediated stimulation of adenylyl cyclase activity was associated with beta 1-adrenoceptors, whereas the remaining 55% was mediated via beta 2-adrenoceptors. The subtype selective antagonists inhibited isoproterenol stimulated aromatization of testosterone to estradiol-17 beta in a concentration-dependent manner. Complete inhibition of beta 1-adrenoceptors resulted in a 45% reduction of estradiol-17 beta formation, whereas similar inhibition of beta 2-adrenoceptors resulted in only a 35% reduction. It is concluded that cAMP-dependent effects of beta-adrenergic agonists in Sertoli cells are mediated by activation of both beta 1- and beta 2 adrenoceptors. The discrepancy between the relative number of beta 1- and beta 2 adrenoceptors and their relative contribution to cAMP production and aromatization indicates that beta 2-adrenoceptors in Sertoli cells are more tightly coupled to the adenylyl cyclase system than beta 1-adrenoceptors. Furthermore, complete inhibition of either beta 1- or beta 2-adrenoceptors by subtype selective antagonists, demonstrates a substantial fraction of spareness between agonist activation of the adenylyl cyclase complex and aromatization. PMID- 8424897 TI - Enzyme immunoassay for plasma 25-hydroxyvitamin D3 employing immunoaffinity chromatography as a pretreatment method. AB - An enzyme immunoassay (EIA) of 25-hydroxyvitamin D3 [25(OH)D3] has been developed as a new methodology for measuring its plasma levels. Three anti-25(OH)D3 antibodies elicited against 25(OH)D3 3-hemisuccinate or -hemiglutarate conjugated with bovine serum albumin were used for this purpose. Two enzyme-labeled antigens were prepared by linking these haptens to beta-galactosidase employing the N succinimidyl ester method. An EIA system, selected from several combinations of the antibodies and labeled antigens, exhibited higher sensitivity and specificity than those of the conventional competitive protein binding assay. However, direct measurement of plasma specimens gave lower values than those obtained from the ones which were pretreated with a Sephadex LH-20 column followed by a normal phase high-performance liquid chromatography. This problem has been overcome by employing immunoaffinity chromatography (IAC). In IAC the homologous anti 25(OH)D3 antibody with that used in the EIA was immobilized as a pretreatment method. The IAC/EIA system developed in this study afforded reliable plasma 25(OH)D3 levels which were confirmed by serial dilution study and the recovery test. The 25(OH)D3 levels of healthy volunteers in the summer measured by this method were 25.2 +/- 6.2 ng/ml (n = 10). PMID- 8424898 TI - Reproducibility of FCM-DNA ploidy analysis in prostatic cancer: comparison between needle biopsy and surgical specimens. AB - Cytofluorometric analysis of FCM-DNA ploidy also seems to have prognostic value in prostatic carcinoma. Determination of the DNA index preoperatively from needle biopsy samples would be advantageous since it would enable the type of intervention to be established in advance. The finding of intratumoural variability in the DNA index in different solid tumours prompted us to compare the data obtained from needle biopsy and surgical specimens of prostatic cancer. Although the results obtained confirmed the heterogeneity of DNA content in prostatic carcinoma, they nonetheless indicate that preoperative DNA analysis is useful, in particular when aneuploidy is observed, since it provides the clinician with additional information on which to base treatment decisions. PMID- 8424899 TI - Flow cytometric assay of lysosome-activated leukocytes as cellular marker of health status. AB - Rapid analysis of the lysosomal system state in human peripheral blood leukocytes was carried out by means of flow cytometry of diluted blood supravitally stained with acridine orange. For each of the subjects studied (26 healthy donors, nine donors with a weak cold-like infection and 30 patients with severe wounds), all the leukocytes were distributed into two distinct cell peaks: one with activated (A) and one with inactivated (I) lysosomal apparatus. It is shown here that whereas the mean cell frequency ratio of the former to the latter peak (A/I) of healthy donors was 1.18 (SD = 0.35) and was consistent with a Gaussian distribution, this value was statistically significantly increased in patients with cold-like symptoms (mean = 2.72, SD = 0.59, P < 10(-5), forming a bimodal distribution when pooled with data for healthy donors, and it increased dramatically (up to 40 times) in patients suffering the aftermath of severe wounds. Therefore, this method, needing only a few drops of the patient's blood and taking 10-15 min, can serve as a marker of health status. In addition, we show that activation/inactivation of the lysosomal apparatus of human leukocytes in vivo proceeds in an 'all-or-none' fashion. It will be possible in the future to enhance the technique by means of evaluating the A/I cell frequency ratio for separate leukocyte types. PMID- 8424900 TI - DNA image cytometry of bladder tumours: comparison of washings and tumour imprints from 61 patients. AB - Cellular DNA cytometry is commonly used to assess the prognosis of bladder tumours. Measurements are made mainly on voided urine or irrigation fluids by flow or image cytometry. In order to determine whether this material is really representative of the bladder tumour, we compared DNA assessments of bladder washings and tumour imprints from the same patients using image cytometry on Feulgen-stained preparations. DNA aneuploidy was found in 41.0% of the washings, 42.6% of the imprints and 49.2% of the patients if both samples were considered. If DNA histograms are classified into three groups (diploid, diploid+aneuploid, aneuploid), a 70% concordance rate is obtained between washings and imprints. In 15% of the cases, an aneuploid population was found in both samples, but in different proportions. In the latter 15%, an aneuploid population was found in one sample but not in the other; this discrepancy, which is probably due to poor tumour exfoliation or to tumour heterogeneity, is of clinical significance since it indicates the importance of making DNA assessments on both washings and cells directly obtained from the tumour. PMID- 8424901 TI - Semi-automated cervical smear pre-screening systems: an evaluation of the Cytoscan-110. AB - This paper reports a test of a system for provision of machine assistance in cervical cytology screening. The hypothesis tested was that if the results of examination by a screener of a small number of high-ploidy cells on specially prepared monolayers, automatically selected and presented by the system, were combined with machine measurement of cell and cell population characteristics, it would be possible to distinguish conditions requiring further action on the part of a cytology service from those in which the patient could safely be signed out. The system appeared broadly capable of this discrimination, with a false-negative error not significantly different (for the numbers tested) on CIN1 and more severe cases to that obtaining for routine screening of the parallel PAP smears, and also to results obtained by a panel of three observers. The machine system appeared to do better than other systems in selecting borderline cases for review, but this may have been an artefact of the method of evaluation used: all results were compared with a 'reference diagnosis', which was computed using statistical techniques to integrate diagnostic information from all available sources. The false-negative error-rate of the system amounted to 5% of high-grade cases, 17% of CIN1's and 29% of borderlines, but were not substantially different from the FN rates for other reporting systems on the same material. The proportion of negative cases referred back for full cytological diagnosis was 34%. Despite this high false-positive rate, the system is potentially cost effective in use. PMID- 8424902 TI - Differentiation of Y79 retinoblastoma cells induced by succinylated concanavalin A. AB - The growth and differentiation potential of Y79 human retinoblastoma cells was assessed in vitro following treatment with the differentiating agent succinylated concanavalin A (SCA). Since SCA treatment induced Y79 cells to display differentiated morphologies in vitro, we sought to determine potential differentiated phenotypes with the use of retinal cell markers. Seventy-two h after SCA treatment, Y79 cells exhibited a decrease in the glial cell marker GFAP and a dramatic and reversible increase in the photoreceptor marker IRBP, while maintaining neuron-specific enolase and PGP 9.5 positivity. These results were indicative of a predominantly neuronal, photoreceptor cell population in response to SCA treatment. In addition, Y79 cell growth inhibition was observed in response to SCA, which could be reversed within 24 h of treatment with the blocking sugar alpha-methyl-D-mannoside. These changes were accompanied by a significant modulation of the N-MYC oncoprotein, as detected by Western blot analysis and immunocytochemistry. Thus, in this system, the status of N-MYC seems to be closely linked to changes in the growth and differentiated state of SCA treated Y79 retinoblastoma cells. PMID- 8424903 TI - Actions and interactions of glucocorticoids and transforming growth factor beta on two related human myeloma cell lines. AB - To evaluate possible involvement of a paracrine/autocrine inhibitory growth factor in myeloma cell growth, we studied the expression and actions of two forms of transforming growth factor beta (TGF-beta 1 and TGF-beta 2) on two closely related myeloma cell lines, OPM-1 and OPM-2. Earlier studies showed that both cell lines contain glucocorticoid receptors, but only OPM-2 cells are growth inhibited by dexamethasone (Dex). We found that OPM-2 growth was inhibited by TGF beta, with TGF-beta 1 exerting a greater effect than TGF-beta 2, and Dex plus TGF beta 1 acting synergistically. In OPM-1 (Dex insensitive), TGF-beta mRNA was not expressed, whereas it was induced by Dex in OPM-2. It was also possible to block partially the growth inhibition of Dex in OPM-2 cells by the addition of anti-TGF beta 1 antibodies. These data suggest that the glucocorticoid effect(s) on myeloma cells may be mediated at least in part through modulation of internal and/or external levels of TGF-beta 1. PMID- 8424904 TI - Extracellular matrix regulation of multidrug resistance in primary monolayer cultures of adult rat hepatocytes. AB - Previous studies reported that, in the absence of drug exposure, multidrug resistance, including resistance to Adriamycin (ADR), could develop in primary rat hepatocyte cultures (B. Carr, Proc. Am. Assoc. Cancer Res., 29:1158, 1988). However, the hepatocytes in that report were cultured on plastic without the benefit of an extracellular matrix (ECM). Because the ECM regulates hepatic gene expression, we have critically evaluated in primary cultures of rat hepatocytes how the ECM affects hepatic ADR resistance, the level of the drug efflux transporter associated with MDR, P-glycoprotein (pgp), and transport of a prototypical pgp substrate, vincristine. Hepatocytes cultured on type I collagen (Vitrogen) had greater resistance to ADR toxicity accompanied by parallel increases in the level of pgp mRNA, decreased drug accumulation, and enhanced drug efflux when compared with the hepatocytes maintained on the basement membrane matrix Matrigel. The development of ADR resistance coincided with the time course of increased pgp mRNA but was not coincident with the time course of expression of either the placental isozyme of glutathione S-transferase or P-450 reductase, proteins associated with MDR in some resistance models. Southern blot analysis revealed neither gross changes in pgp gene structure or gene copy number to account for the increase in pgp RNA levels for hepatocytes cultured on Vitrogen. ECM also regulated xenobiotic-inducible expression of hepatic pgp, since chemotherapeutic agents, including vincristine and colchicine, induced pgp mRNA exclusively in hepatocytes cultured on Vitrogen. The critical matrix proteins in Matrigel responsible for regulation of pgp were determined by the selective addition of its components to the culture environment. The presentation of the individual matrix elements as a rigid substratum to the hepatocyte did not decrease pgp mRNA. In contrast, the presentation to the same hepatocytes of either laminin or type IV collagen in a nonrigid state (solubly in the medium) selectively decreased hepatocellular pgp mRNA. We conclude that primary rat hepatocytes develop ADR resistance with time in culture due to increased expression of pgp and that ECM proteins represent endogenous physiological modulators of both basal and chemotherapeutically inducible expression of hepatic P-glycoprotein. PMID- 8424905 TI - Regulation by bcl-2, c-myc, and p53 of susceptibility to induction of apoptosis by heat shock and cancer chemotherapy compounds in differentiation-competent and defective myeloid leukemic cells. AB - Myeloid leukemias that differ in their competence for induction of differentiation were analyzed for expression of bcl-2 and c-myc and for their sensitivity to induction of apoptosis by heat shock and cancer chemotherapy compounds. The M1 leukemia expressed a high level of bcl-2 and showed a much lower susceptibility to induction of apoptosis by heat shock, Adriamycin, 1-beta D-arabinofuranosylcytosine, methotrexate, and cycloheximide, compared to five other leukemias which expressed a low level of bcl-2. There was no association between susceptibility to induction of apoptosis and competence for induction of differentiation. The difference in susceptibility to methotrexate, which is not regulated by the multidrug resistance (MDR) genes, and treatment with verapamil, which blocks MDR activity, have indicated that the higher resistance of the M1 leukemia to these agents was not due to MDR activity. The results indicate that the level of regulated bcl-2 expression in these myeloid leukemias was associated with cell susceptibility to induction of apoptosis by different apoptosis inducing agents. Screening for expression of bcl-2 may thus be useful to characterize leukemias regarding susceptibility to induction of apoptosis by different agents. The level of regulated c-myc expressed in these leukemias was not associated with susceptibility to induction of apoptosis. Transfection with a deregulated mutant p53 into the M1 leukemia did not change susceptibility to apoptosis induction, but transfection with deregulated c-myc increased susceptibility to apoptosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424906 TI - Consequences of heteromeric interactions among helix-loop-helix proteins. PMID- 8424907 TI - Ha-ras and v-raf oncogenes, but not int-2 and c-myc, interfere with the lactogenic hormone dependent activation of the mammary gland specific transcription factor. AB - beta-Casein milk protein expression is regulated synergistically by the lactogenic hormones dexamethasone, insulin, and prolactin. This regulation has been observed in vivo and in the mammary epithelial cell line HC11. An important mediator of hormone action on beta-casein gene transcription is the mammary gland specific transcription factor MGF. Epidermal growth factor is a potent growth promoter for HC11 cells and causes a state of predifferentiation in confluent HC11 cells. Epidermal growth factor receptor activation, however, antagonizes the effects of the lactogenic hormones on MGF activation and beta-casein gene transcription during the induction of predifferentiated cells. We examined the effects of two downstream components of the epidermal growth factor receptor mitogenic signaling pathway on MGF and beta-casein promoter activity. Constitutively activated Ha-ras or v-raf oncogenes were introduced into HC11 cells. HC11 cell lines transfected with Ha-ras or v-raf oncogenes assumed transformed properties and were blocked in the lactogenic induction of MGF and beta-casein gene transcription. This indicates that the same components instrumental in the mitogenic pathway are also involved in the block of differentiation specific gene expression in HC11 cells. This block is not due to the transformed state of the HC11 cells. Introduction and high expression of the int-2 gene, a member of the fibroblast growth factor gene family, does not interfere with MGF and beta-casein induction. Overexpression of the c-myc gene, causing mammary epithelial cell transformation, also does not suppress MGF activity.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424908 TI - The West Midlands epithelial ovarian cancer adjuvant therapy trial. AB - In a multicentre prospective randomized controlled trial, single agent cisplatinum was compared with whole abdomino-pelvic moving strip radiotherapy in the management of Stage IC-III epithelial ovarian cancer patients who had no macroscopic residual disease after primary surgery. Over a 6-year period 40 eligible patients were recruited, 15 of whom had Stage III disease. The overall 5 year survival was 60% with no significant survival difference between the treatment groups. Acute toxicity was common in both arms and six (11%) patients experienced significant long term disability. PMID- 8424909 TI - The efficacy and safety of ondansetron in the prophylaxis of cancer-chemotherapy induced nausea and vomiting in children. AB - The management of chemotherapy induced emesis presents a major difficulty in the paediatric population. Ondansetron is a 5-HT3 antagonist and its antiemetic properties have been established in adults receiving chemotherapy. Experience in the treatment of children, however, is limited. There is no standard comparative antiemetic in paediatric practice, so this European, multi-centred study aimed to assess the clinical efficacy and safety of ondansetron in a large cohort of children receiving a range of emetogenic chemotherapy regimens for malignant disease. Two hundred patients were entered into the study, of whom 183 fulfilled entry criteria. Forty per cent (10/25) of patients receiving cisplatin, 68% (27/40) receiving ifosfamide and 70% (83/118) receiving other drugs had less than 3 emetic episodes (vomit or retch) during their worst 24 hours of chemotherapy. The number of days without vomiting or retching during and shortly after receiving chemotherapy was also related to the emetic potential of the agents in the regimen: cisplatin 82/182 (45%), ifosfamide 137/213 (64%) and other agents 430/566 (76%). The control of nausea appeared better than that of emesis in each of the sub-groups analysed. Ondansetron was safe, well tolerated and effective in the prevention of vomiting in children receiving a wide variety of chemotherapy regimens. PMID- 8424910 TI - A randomized trial to assess the efficacy of 5-aminosalicylic acid for the prevention of radiation enteritis. AB - Sulphasalazine is an effective treatment for diarrhoea occurring during pelvic radiotherapy. We report the results of a trial to assess the value of its active moiety, 5-aminosalicylic acid, (5-ASA) in a prophylactic setting. Seventy-three patients planned for external beam radiotherapy to the pelvis were randomized on a double-blind basis to receive prophylactic 5-ASA or placebo. The severity of the acute radiation bowel reaction was documented by a weekly questionnaire. Surprisingly, diarrhoea occurred in a higher proportion of patients in the 5-ASA arm than the placebo arm (91.2% versus 73.7%, P = 0.070). The maximum change in both the severity of diarrhoea and the number of days per week on which diarrhoea occurred (from pre-radiotherapy level to the worst level at any time during treatment) were both significantly greater in patients taking 5-ASA than those taking placebo (P = 0.014 and P = 0.026, respectively). The average change (the sum of the weekly scores divided by the number of weeks of treatment, minus the pre-radiotherapy score) for both severity and days per week of diarrhoea were again greater in the 5-ASA than the placebo arm, but failed to reach statistical significance (P = 0.095 and P = 0.079, respectively). The use of anti-diarrhoeal medicines was significantly greater in the 5-ASA arm (P = 0.011). Constipation was more common in the placebo arm but this did not reach significance (P = 0.20). 5-ASA thus has no protective effect against acute radiation enteritis and appears to worsen it. Possible reasons for this surprising finding are discussed. PMID- 8424911 TI - A phase II study of epirubicin in advanced transitional cell bladder cancer. The Yorkshire Urological Cancer Research Group. AB - A multicentre Phase II study of epirubicin has been performed in patients with measurable or evaluable recurrent or metastatic transitional cell bladder cancer. Epirubicin was given intravenously every 3 weeks at a dose of 100 mg/m2. An objective response rate of 28% was observed (one complete and nine partial remissions) in an evaluable group of 36 patients, (confidence interval 15%-45%). Subjective improvements in the condition of patients were seen in responding and 'no change' patients. An interesting observation was the good response seen in patients with recurrent bladder disease who had previously received radial radiotherapy. Toxicity was considered to be acceptable and manageable, the most frequent being alopecia, and nausea and vomiting. Haematological toxicity was slight. One patient developed skin pigmentation, a not previously recognized complication of epirubicin treatment. Three cases of possible cardiotoxicity were seen in the 43 patients evaluable for toxicity. Epirubicin in thus an active agent in transitional cell carcinoma of the bladder, with a role as a single agent for palliation. It may also be useful in combination regimens for more aggressive treatment. PMID- 8424912 TI - Changes in target volume during radiotherapy treatment of invasive bladder carcinoma. AB - Ninety patients with stage T3 Nx Mo carcinoma of the urinary bladder were treated with radical megavoltage external beam radiotherapy. Planning for treatment was undertaken on a treatment planning system utilizing CT scan slices to define the target volume and patient outline. All patients underwent a second CT scan half way through their course of treatment to assess any change in target volume and the continued adequacy of the original treatment plan. Seventy-two patients (80%) had no spatial shift in target volume, but, of the 18 patients with such a shift, treatment plans were changed in seven. The majority of patients had no delay in continuing their treatment after replanning, but one patient had a gap of 5 days before restarting treatment. An analysis of the factors possibly associated with a change in target volume showed that a primary tumour at the bladder base, rather than elsewhere in the bladder, was the single most important criterion for predicting target volume changes. There was no correlation between the size of the initial tumour, or the size of the prostate gland in male patients, and the occurrence of a shift in volume outside the initial target volume. Some method of regularly assessing the continued relevance of the target volume may be needed in this group of patients to improve the precision of treatment and also improve results. PMID- 8424913 TI - Fractionated total body irradiation: some radiobiological considerations. AB - Fractionated total body irradiation (TBI), as conditioning regimen for bone marrow transplantation in the treatment of non-lymphocytic leukaemia, has proved to be less toxic than single fraction TBI. However, higher relapse rates are reported. The linear quadratic (LQ) model has been applied to calculate equivalent total therapeutic doses as well as equivalent tolerance doses for fractionated TBI. The LQ model predicted that escalating total doses for fractionated TBI will still result in acceptable toxicity to lung while achieving better disease control. There is published clinical data supporting our conclusion from this radiobiological analysis. PMID- 8424914 TI - Carboplatin and 5-fluorouracil in advanced and recurrent squamous carcinoma of the head and neck. AB - Twenty-six patients with advanced squamous carcinoma of the head and neck or local recurrence after surgery and/or radiotherapy received carboplatin 300 mg/m2 intravenously on day 1 and 5-fluorouracil 1 gm/m2 by continuous intravenous infusion for 4 days. The treatment was well tolerated with little toxicity. The overall response rate was 58%. PMID- 8424915 TI - Response of metastatic anal carcinoma to single agent carboplatin. AB - The management of locoregional squamous cell carcinoma of the anus with a combined modality approach comprising chemotherapy and radiotherapy is well established. However, the optimum regimen for the management of metastatic disease has yet to be determined. Cisplatin has been shown to have some efficacy in this disease. We report a case of partial response of metastatic disease to single agent carboplatin, and discuss its role in this situation. PMID- 8424916 TI - Multiple myeloma presenting as an acute abdomen. AB - We report a case of myeloma which presented as an acute abdomen with an associated pancreatic mass. This clinical picture has not been reported previously. PMID- 8424917 TI - Pelvic failure rate and radiation toxicity in relation to total dose of radiation alone for the treatment of cancer of the uterine cervix. AB - Seventy-four patients presenting between 1978 and 1989 with carcinoma of the uterine cervix were analysed to compare our results with the published literature, especially with regard to the pelvic failure rate and radiation toxicity. Overall 5-year survival reached 41%. Overall 2-year survival rate for Stage IIB was 85% and Stage IIIB was 32%. For Stage IIB the results agreed with those reported in the literature. Pelvic control was related to total dose and the application of intracavitary irradiation. For Stage IIIB the results were disappointing, partly due to a large number of bulky tumours and also to a relatively low paracentral dose and dose to the pelvic side wall. Percentages of radiation side-effects were low compared with those reported in the literature, probably also due to the relatively low total dose. PMID- 8424918 TI - Pathological fracture in haemoglobinopathy: treatment by irradiation. AB - A patient suffering from thalassaemia, with extreme osteoporosis, coarse trabeculation and cortical thinning of the bones, developed a pathological fracture of the left hip. This was treated by a single dose of radiotherapy. It is suggested that the radiotherapy facilitated the healing process by eliminating the causative factor of the fracture, which was the expanding and over proliferating bone marrow. PMID- 8424919 TI - Unilateral hyperhidrosis associated with intrathoracic IgD lambda myelomatous tumour. AB - Localized sympathetic over-activity is a rare manifestation of intrathoracic malignancy. We report the case of a 62-year-old woman who presented with a right sided hyperhidrosis limited to the face, trunk and upper extremities; this revealed a left paraspinal intrathoracic IgD lambda myelomatous tumour. The episodes of sweating were abolished by local radiotherapy, concomitant with the disappearance of the tumour. PMID- 8424920 TI - Recoverin immunoreactivity in mammalian cone bipolar cells. AB - Human, macaque monkey, and rat retinas were immunostained with a polyclonal antibody preparation against purified recoverin, a 23-kD calcium-binding protein isolated from bovine retina that localizes to rods and cones (Dizhoor et al., 1991). In addition to immunoreactive photoreceptors, we have identified subpopulations of recoverin-positive bipolar cells in all three species. Results from immunostaining with progressive dilutions of anti-recoverin and preadsorption of the antibody with a dilution series of purified recoverin showed that photoreceptors and bipolar cells had similar affinities for the antibody and suggested that the molecule recognized by the antibody in both cell types is recoverin. Immunoreactivity for recoverin and protein kinase C, a selective marker for all rod bipolar cells, was found in separate bipolar cell populations. Recoverin immunoreactivity is therefore a characteristic of certain cone bipolar cell types. In rat retina, anti-recoverin labeled two morphologically distinct subpopulations of cone bipolar cells whose axonal arbors stratified at different depths in the inner plexiform layer (IPL). The bipolar cells labeled with anti recoverin did not correspond to those that were reactive for calbindin, another cone bipolar cell marker. Human and monkey retinas also had two populations of cone bipolar cells that were recoverin-positive. One population showed a distinct pattern of narrow bistratification at the outer border of the IPL and a regular mosaic arrangement of its axonal arbors, suggesting that the entire population of a single cone bipolar type was labeled. Cell density, dendritic morphology, and axonal-field size and stratification indicate that anti-recoverin selectively strains the flat midget (presumed OFF-center) cone bipolar cell type observed previously in Golgi preparations. By contrast the second bipolar cell population had axonal stratification in the inner half of the IPL and showed an unusual but consistent morphology and spatial distribution. Individual cells were intensely stained but were present at an extremely low density (approximately 2-5 cells/mm2). These cells had multibranched dendritic trees characteristic of the diffuse bipolar cell class, but very small axonal fields in the size range of the midget bipolar class. Neither of the two recoverin-positive bipolar cell types in monkey was labeled with anti-calbindin or anti-cholecystokinin. An antibody preparation against bovine pineal hydroxyindole-O-methyltransferase (HIOMT) labeled photoreceptors and bipolar cells that closely resembled the recoverin positive bipolar cells in human and rat retinas.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8424921 TI - Prenatal development of axon outgrowth and connectivity in the ferret visual system. AB - The objective of this study was to determine when the retina, lateral geniculate nucleus (LGN), and striate cortex first send out axons, and first connect with each other, during embryonic development in the ferret. Specifically, we were interested in the timing relationship between axon outgrowth and known temporal patterns of neurogenesis in the LGN and striate cortex. Ferrets (Mustela putorius furo) were selected for study because of their immature developmental state in late gestation and relatively large litters. We examined axon outgrowth from the retina, and anlagen of presumptive LGN and striate cortex between embryonic day 21-30 (E21-E30) using in situ inoculations of two fluorescent lipophilic dyes, DiI and DiA. DiI inoculations were made into the cortex and contralateral thalamus, and DiA inoculations were made into the contralateral eye. Retinal axon termination zones in the diencephalon following the DiA inoculations were used to validate the location of the LGN. Visual cortex and LGN neurogenesis begins at E20 in ferrets. No axon outgrowth could be documented from retina or anlagen of striate cortex and LGN until E24. At E24 some retinal axons reach and cross the chiasm, cortical axons extend some distance within the cortical radiations, and thalamic axons are within the internal capsule. Retinogeniculate, geniculocortical, and corticogeniculate axons extend to their target structures by E27, as evidenced by retrograde labeling in cells of origin. These data suggest that in the ferret retina, and developing LGN and striate cortex, (1) axon outgrowth from each visual area begins within 24-h of each other, after neurogenesis has begun at the source but before it is complete in the target; (2) axons may be generated before parent cell bodies have completed migration; and (3) arriving axons are in a position to influence target structures almost from their inception. PMID- 8424922 TI - Temporal asynchrony interferes with vernier acuity. AB - Two dots may be aligned vertically with a precision much higher than that expected from two-point resolution provided they are separated by a visual angle of 3-5 min of arc. This precision suffers when the two dots are not exposed synchronously. Neither onset nor offset asynchronies can be tolerated; exposure differences of the two components of the vernier task as low as 30 ms can lead to a reduction in performance when the total exposure is below 90 ms. This effect cannot be compensated for by synchronizing the onset of one stimulus component with the offset of the other, even when the two are of opposite contrast. The data suggest that vernier acuity may be subserved by a dynamical linking of cortical excitation generated by the synchronous arrival of signals within a range of locations in the cortex whose spatial separation is critical for optimal hyperacuity performance. The evidence presented in this paper must be taken into account when a physiological substrate for hyperacuity is considered. PMID- 8424923 TI - A role for the corpus callosum in visual area V4 of the macaque. AB - The classically defined receptive fields of V4 cells are confined almost entirely to the contralateral visual field. However, these receptive fields are often surrounded by large, silent suppressive regions, and stimulating the surrounds can cause a complete suppression of response to a simultaneously presented stimulus within the receptive field. We investigated whether the suppressive surrounds might extend across the midline into the ipsilateral visual field and, if so, whether the surrounds were dependent on the corpus callosum, which has a widespread distribution in V4. We found that the surrounds of more than half of the cells tested in the central visual field representation of V4 crossed into the ipsilateral visual field, with some extending up to at least 16 deg from the vertical meridian. Much of this suppression from the ipsilateral field was mediated by the corpus callosum, as section of the callosum dramatically reduced both the strength and extent of the surrounds. There remained, however, some residual suppression that was not further reduced by addition of an anterior commissure lesion. Because the residual ipsilateral suppression was similar in magnitude and extent to that found following section of the optic tract contralateral to the V4 recording, we concluded that it was retinal in origin. Using the same techniques employed in V4, we also mapped the ipsilateral extent of surrounds in the foveal representation of V1 in an intact monkey. Results were very similar to those in V4 following commissural or contralateral tract sections. The findings suggest that V4 is a central site for long-range interactions both within and across the two visual hemifields. Taken with previous work, the results are consistent with the notion that the large suppressive surrounds of V4 neurons contribute to the neural mechanisms of color constancy and figure-ground separation. PMID- 8424924 TI - Photopigments of dogs and foxes and their implications for canid vision. AB - Electroretinogram (ERG) flicker photometry was used to examine the photopigment complements of representatives of four genera of Canid: domestic dog (Canis familiaris), Island gray fox (Urocyon littoralis), red fox (Vulpes vulpes), and Arctic fox (Alopex lagopus). These four genera share a common cone pigment complement; each has one cone pigment with peak sensitivity of about 555 nm and a second cone pigment with peak at 430-435 nm. These pigment measurements accord well with the conclusions of an earlier investigation of color vision in the dog, and this fact allows some predictions about color vision in the wild canids. An additional set of measurements place the peak of the dog rod pigment at about 508 nm. PMID- 8424925 TI - Effects of vasoactive intestinal peptide on ganglion cells in the rabbit retina. AB - The effects of vasoactive intestinal peptide (VIP) on the extracellularly recorded activity of ganglion cells were studied in superfused rabbit retinas. VIP, applied to the bathing solution, significantly increased the maintained activity of both OFF-center and ON-center brisk ganglion cells. The transient, excitatory responses of these cells to flashes of light (spots or annuli centered over the receptive field) were either unaffected or moderately reduced. VIP did not affect the responses of most ON/OFF directionally selective ganglion cells to a moving light stimulus. Furthermore, maintained activity of most of these cells remained absent. Effects of VIP on the light responses of ganglion cells were pronounced in retinas that were bathed with the dopamine D1 antagonist (+)-SCH 23390. For OFF-center brisk ganglion cells, VIP brought out both the center and surround excitatory responses that were selectively reduced by (+)-SCH 23390. Similarly, VIP brought out the leading edge responses that were reduced by (+) SCH 23390 in ON/OFF directionally selective ganglion cells to a moving light stimulus. VIP did not however reverse the effects of (+)-SCH 23390 on ON-center brisk ganglion cells. It is argued that VIP counteracts the effects of (+)-SCH 23390 on OFF-center brisk ganglion cells and ON/OFF directionally selective ganglion cells by stimulating adenylate cyclase activity in dopamine-receptive cells of the retina. PMID- 8424926 TI - Visual processing in pigeon nucleus rotundus: luminance, color, motion, and looming subdivisions. AB - The responses of single cells to luminance, color and computer-generated spots, bars, kinematograms, and motion-in-depth stimuli were studied in the nucleus rotundus of pigeons. Systematic electrode penetrations revealed that there are several functionally distinct subdivisions within rotundus where six classes of visual-selective cells cluster. Cells in the dorsal-posterior zone of the nucleus respond selectively to motion in depth (i.e. an expanding or contracting figure in the visual field). Most cells recorded from the dorsal-anterior region responded selectively to the color of the stimulus. The firing rate of the cells in the anterior-central zone, however, is dramatically modulated by changing the level of illumination over the whole visual field. Cells in the ventral subdivision strongly respond to moving occlusion edges and very small moving objects, with either excitatory or inhibitory responses. These results indicate that visual information processing of color, ambient illumination, and motion in depth are segregated into different subdivisions at the level of nucleus rotundus in the avian brain. PMID- 8424927 TI - Response properties of ganglion cells in the isolated mouse retina. AB - We have studied the organization of receptive fields of ganglion cells in the isolated mouse retina and have shown that the organization is similar to that of the cat. Based upon responses to circular and annular stimuli, most ganglion cells (90%; N = 83) had receptive fields with concentric center-surround organization, either ON or OFF center. The plot of response amplitude vs. stimulus area for these cells increased to a maximum (corresponding to a diameter of 10.0 +/- 2.8 deg S.E.M.; N = 13) and then decreased for larger stimuli, demonstrating the presence of an antagonistic surround. The dark-adapted sensitivity (205 +/- 43.8 impulses quantum-1 rod-1; mean +/- S.E.M.) did not differ from that measured for the intact preparation. We found a subset of OFF center cells for which the dark discharge was very regular (mean coefficient of variation = 0.30). Using sinusoidal grating stimuli, we classified ganglion cells as X-like (87%) and Y-like (13%) based on spatial summation properties and the presence of subunit activity in the receptive-field center. We found no difference in the spatial-frequency preference between X-like and Y-like cells in the central retina (high cutoff frequency, 0.20 +/- 0.014 cycle/deg, mean +/- S.E.M.), in contrast to the marked difference between X cells and Y cells in the cat. Thus, ganglion cell receptive fields in the mouse retina resemble those of the cat, although the spatial characteristics of the receptive fields in the central retina are more homogeneous. This homogeneity would simplify the comparison of retinas from normal and mutant strains of the mouse. PMID- 8424928 TI - Comparison of subcortical connections of inferior temporal and posterior parietal cortex in monkeys. AB - To investigate the subcortical connections of the object vision and spatial vision cortical processing pathways, we injected the inferior temporal and posterior parietal cortex of six Rhesus monkeys with retrograde or anterograde tracers. The temporal injections included area TE on the lateral surface of the hemisphere and adjacent portions of area TEO. The parietal injections covered the posterior bank of the intraparietal sulcus, including areas VIP and LIP. Our results indicate that several structures project to both the temporal and parietal cortex, including the medial and lateral pulvinar, claustrum, and nucleus basalis. However, the cells in both the pulvinar and claustrum that project to the two systems are mainly located in different parts of those structures, as are the terminals which arise from the temporal and parietal cortex. Likewise, the projections from the temporal and parietal cortex to the caudate nucleus and putamen are largely segregated. Finally, we found projections to the pons and superior colliculus from parietal but not temporal cortex, whereas we found the lateral basal and medial basal nuclei of the amygdala to be reciprocally connected with temporal but not parietal cortex. Thus, the results show that, like the cortical connections of the two visual processing systems, the subcortical connections are remarkably segregated. PMID- 8424929 TI - Processing of form and motion in area 21a of cat visual cortex. AB - Extracellular recordings from single neurons have been made from presumed area 21a of the cerebral cortex of the cat, anesthetized with N2O/O2/sodium pentobarbitone mixture. Area 21a contains mainly a representation of a central horizontal strip of contralateral visual field about 5 deg above and below the horizontal meridian. Excitatory discharge fields of area 21a neurons were substantially (or slightly but significantly) larger than those of neurons at corresponding eccentricities in areas 17, 19, or 18, respectively. About 95% of area 21a neurons could be activated through either eye and the input from the ipsilateral eye was commonly dominant. Over 90% and less than 10% of neurons had, respectively, C-type and S-type receptive-field organization. Virtually all neurons were orientation-selective and the mean width at half-height of the orientation tuning curves at 52.9 deg was not significantly different from that of neurons in areas 17 and 18. About 30% of area 21a neurons had preferred orientations within 15 deg of the vertical. The mean direction-selectivity index (32.8%) of area 21a neurons was substantially lower than the indices for neurons in areas 17 or 18. Only a few neurons exhibited moderately strong end-zone inhibition. Area 21a neurons responded poorly to fast-moving stimuli and the mean preferred velocity at about 12.5 deg/s was not significantly different from that for area 17 neurons. Selective pressure block of Y fibers in contralateral optic nerve resulted in a small but significant reduction in the preferred velocities of neurons activated via the Y-blocked eye. By contrast, removal of the Y input did not produce significant changes in the spatial organization of receptive fields (S or C type), the size of the discharge fields, the width of orientation tuning curves, or direction-selectivity indices. Our results are consistent with the idea that area 21a receives its principal excitatory input from area 17 and is involved mainly in form rather than motion analysis. PMID- 8424930 TI - Chemical modification of alpha-subunit tryptophan residues in Schizosaccharomyces pombe mitochondrial F1 adenosine 5'-triphosphatase: differential reactivity and role in activity. AB - Chemical modification of mitochondrial F1-ATPase from Schizosaccharomyces pombe by the tryptophan-specific reagent N-bromosuccinimide (NBS) at pH 5.0 in the presence of 20% glycerol produced a characteristic lowering in both enzyme absorbance at 280 nm and intrinsic fluorescence at 332 nm that varied with NBS/F1 molar ratio up to a value of 130. Fluorometric titration of tryptophans and correlation to residual ATPase activity showed that modification of three reactive residues among the seven present on alpha- and epsilon-subunits did not markedly modify the enzyme activity but efficiently released endogenous ATP and abolished the fluorescence quenching related to GDP or ATP binding to the catalytic site. Additional modification of one, less reactive, tryptophan altered both negative cooperativity of ATP hydrolysis and sensitivity to azide inhibition and produced a nearly complete inactivation at high NBS/F1 molar ratio. NBS induced inactivation of F1 was favored by catalytic-site saturation with GDP or low ATP concentration and on the contrary was prevented by noncatalytic-site saturation with ADP or high ATP concentration. When reactive tryptophans were selectively modified by NBS in the presence of ADP, and subunits were isolated after guanidine hydrochloride dissociation by one-step purification on reversed phase HPLC, the absorbance of alpha-subunit at 280 nm was decreased, whereas that of epsilon-subunit was unchanged. Cyanogen bromide cleavage of alpha-subunit and fragments separation by reversed-phase HPLC showed that one peptide of 3 kDa apparent molecular mass had decreased absorbance. N-Terminal sequencing allowed its identification to fragment 255-282 that contains tryptophan257. PMID- 8424931 TI - Catabolism of adenine nucleotides and its relation with intracellular phosphorylated metabolite concentration during ethanol oxidation in perfused rat liver. AB - Ethanol-induced perturbations in the energy metabolism and in the catabolism of adenine nucleotides were investigated by 31P NMR spectroscopy and HPLC analyses in perfused rat liver. Ethanol oxidation reduced the redox potential of the hepatocyte, leading to an intracellular accumulation of sn-glycerol 3-phosphate. This accumulation, in turn, led to a cytosolic P(i) depletion with stoichiometric relationship close to 1/1 for an initial period of 2 min. The concentration of nucleoside 5'-triphosphates (83 +/- 4% of ATP) was decreased during ethanol oxidation, reaching about 66% of its control value [2.88 +/- 0.02 mumol.(g of liver wet wt)-1] at high ethanol doses (10 and 70 mM). The depletion of P(i) relieved the inhibition exerted by this compound on AMP deaminase, key enzyme in the catabolism of adenine nucleotides. The degradation of AMP was monitored by HPLC analyses of the adenine nucleosides and bases released in the effluents. Integration over time of the total release of these metabolites accounted for the depletion of ATP recorded in the same time by 31P NMR spectroscopy. This result suggests that ATP depletion occurring during ethanol oxidation originated from an enhanced degradation of adenine nucleotides. There was a strong linear correlation (r2 = 0.92) between cytosolic P(i) level and allantoin release rate during ethanol perfusion. Cytosolic P(i) and allantoin release exhibited biphasic behavior, the recovery toward the initial levels being related to the release of P(i) in the cytoplasm during the complete catabolism of adenine nucleotides. Finally, the depletion of P(i) affected the glycogenolysis pathway, with a maximal inhibition of ca. 19% of the initial level. PMID- 8424932 TI - Studies with synthetic peptide substrates derived from the neuronal protein neurogranin reveal structural determinants of potency and selectivity for protein kinase C. AB - The neuronal protein neurogranin, also known as RC3, is a selective substrate for protein kinase C (PKC). We synthesized a peptide corresponding to the phosphorylation domain of neurogranin (amino acids 28-43) and characterized its properties as a PKC substrate. Neurogranin(28-43) was phosphorylated by purified PKC with a Km of 150 nM. No significant phosphorylation of the peptide by either cAMP-dependent protein kinase or by calcium/calmodulin-dependent protein kinase II could be detected. Thus, neurogranin(28-43) is a potent and selective substrate for PKC. We tested several peptide analogues of neurogranin(28-43) for their substrate potency and specificity as kinase substrates, in order to help elucidate the structural determinants involved in the phosphorylation of substrates by PKC. Substituting Arg36 with Ile caused a significant reduction in the affinity for PKC. Replacing Lys30 with Arg enhanced the catalytic efficiency (Vmax/Km) for PKC but diminished the selectivity of the substrate for PKC. These results support the generally held model that basic amino acids on both sides of the phosphorylated Ser are important structural determinants in PKC substrates. However, the data also suggest that the presence of particular basic amino acids (Arg vs Lys) can contribute to the degree of selectivity of a substrate for PKC. Replacement with Ala of Phe35, the amino acid adjacent to the Ser34 phosphorylation site, resulted in a peptide with greatly diminished potency as a PKC substrate. This finding indicates a critical role of Phe35 in modulating binding and phosphorylation of neurogranin-derived peptides by PKC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424933 TI - Formation of the extracellular mucous matrix of olfactory neuroepithelium: identification of partially glycosylated and nonglycosylated precursors of olfactomedin. AB - Olfactomedin is the major glycoprotein of the extracellular mucous matrix of frog olfactory neuroepithelium. It is responsible for the primary architecture of this extracellular matrix by forming via intermolecular disulfide bonds polymers, which are covered with evenly spaced carbohydrate groups. To study glycosylation of olfactomedin, we raised antibodies against the mature protein and antibodies against a region adjacent to an N-linked glycosylation site near its amino terminus. The latter antibodies cannot bind when this site is glycosylated and reveal precursors of olfactomedin in the perinuclear regions of Bowman's glands. In contrast, antiserum against the mature protein stains acinar regions of glands and the ciliary surface. Enzymatic deglycosylation of olfactomedin shows stepwise removal of carbohydrate and reveals a 51-kDa deglycosylated form. Our results indicate that, prior to secretion, most, if not all, of the six potential N linked glycosylation sites of olfactomedin are glycosylated with carbohydrate moieties of about 8-10 sugar residues. PMID- 8424934 TI - Phospholipase C-promoted membrane fusion. Retroinhibition by the end-product diacylglycerol. AB - The catalytic activity of phospholipase C induces fusion of pure lipid vesicles. When large unilamellar liposomes composed of phosphatidylcholine/phosphatidylethanolamine/cholesterol (2:1:1 mole ratio) are treated with phospholipase C, in the presence of 10 mM Ca2+, two enzyme effects can be distinguished: a fast one (half-time on the order of seconds) consisting mainly of vesicle-vesicle fusion and a slow one (half-time on the order of minutes) representing bulk lipid hydrolysis. The fast fusion process is inhibited by the end-product diacylglycerol, as well as by lysophosphatidylcholine and by low Ca2+ concentrations. The temperature dependence of enzyme activity (phospholipid hydrolysis), vesicle aggregation, and vesicle fusion (mixing of aqueous contents) has been separately studied. Enzyme activity and vesicle aggregation rates increase monotonically with temperature, while an optimum temperature is found for vesicle fusion, depending on liposome composition and assay conditions. The presence of diacylglycerol incorporated to the membrane (up to 10 mol %) does not produce any fusion effect even at temperatures as high as 80 degrees C: in situ diacylglycerol production by the enzyme appears to be required. The data are interpreted in support of a hypothesis according to which a "fusion intermediate" would be required, depending (among others) on bilayer composition, temperature, and Ca2+ concentration, for vesicle fusion to occur. PMID- 8424935 TI - Concerted DNA recognition and novel site-specific alkylation by duocarmycin A with distamycin A. AB - Duocarmycin A, a novel antitumor antibiotic, has a reactive cyclopropane ring, which has been reported to alkylate adenine at the 3' end of sequences of three or more consecutive A or T in DNA [Boger, D. L., et al. (1990) J. Am. Chem. Soc. 112, 8961-8971]. In order to study the DNA recognition, the reaction of DNA with duocarmycin A was performed in the presence of DNA ligands. Distamycin A, berenil, Hoechst 33258, and 4',6-diamidino-2-phenylindole (DAPI), which are minor groove binders with affinity to A.T-rich sequences, were used. DNA-sequencing experiments showed that treatment of DNA with duocarmycin A plus distamycin A caused alkylation of guanine residues in G.C-rich sequences, which are not alkylated by duocarmycin A alone. Guanine alkylation by duocarmycin A was not observed with berenil, Hoechst 33258, or DAPI. HPLC product analysis showed that duocarmycin A reacted with a double-helical DNA octamer d(CCCCGGGG)2 in the presence of distamycin A to produce duocarmycin A-guanine adduct, while duocarmycin A alone did not react with the octamer. Chromomycin A3, which binds as a Mg(II)-coordinated dimer to G.C-rich sequences in the minor groove, inhibited the guanine alkylation by duocarmycin A in the presence of distamycin A. A footprinting experiment showed that there is a distamycin A-binding site close to the alkylated guanine residue. These results suggest that two different molecules, duocarmycin A and distamycin A, cooperatively recognize DNA sequences including consecutive G.C base pairs resulting in alkylation at the novel guanine sites. The cooperative drug recognition can be designated as "concerted DNA recognition". PMID- 8424936 TI - An A-form of poly(amino2dA-dT).poly(amino2dA-dT) induced by polyamines. AB - The effect of the naturally occurring polyamines spermidine and spermine on poly(amino2-dA-dT).poly(amino2dA-dT) conformation has been studied by UV, CD, and IR spectroscopies. It is shown that a conformational transition is induced in poly(amino2dA-dT).poly(amino2dA-dT) by micromolar concentrations of the polyamines (30 microM) in low-salt aqueous solution. The analysis of our results, in view of previously published studies on conformational properties of the amino polynucleotide, indicates the resulting conformer to be an A-form. Interestingly, the polyamine concentration at the midpoint of the transition is the same in both cases. This provides further evidence that the coordination of positively charged counterions to DNA is determined largely from the DNA structure, probably with an important role for the sequence, and less from the nature of the ions. PMID- 8424937 TI - Plasmid dimerization mediated by triplex formation between polypyrimidine polypurine repeats. AB - The ability of independent pyr.pur tracts to participate in triplex formation has been investigated in linear plasmids. The pyr.pur tract could be positioned at the ends of the plasmids or internally by a suitable choice of restriction enzyme. Dimer formation between plasmids was monitored by mobility shifts on agarose gels as well as by direct visualization in the electron microscope. Linear dimers and X and Y structures were observed. Control experiments showed that a pyr.pur tract was essential and was consistent with triplex formation in which the two pyrimidine strands were antiparallel. These structures were formed at pHs between 4 and 6, but once formed they remained stable up to pH 7. Spermine was required for formation of dimers at low ionic strength, but once formed the dimers remained stable in the absence of spermine. Additional linear plasmids were constructed with pyr.pur tracts at both ends; these formed structures at pH 4 which had mobilities identical to those of open circles. Triplex formation of this type may serve as a good model for loop formation in eukaryotic chromosomes. PMID- 8424938 TI - The conformation of loop E of eukaryotic 5S ribosomal RNA. AB - The solution structure of a 27-nucleotide duplex, including the internal loop E from Xenopus laevis 5S ribosomal RNA, has been studied by two-dimensional NMR spectroscopy, followed by restrained molecular dynamics. The highly conserved internal loop closes to form a G.A base pair and a reverse-Hoogsteen A.U base pair. Extensive interstrand stacking between these uncommon base pairs provides a structural explanation for an interstrand ultraviolet-induced cross-link. A guanosine residue is bulged into the major groove and may form a base-triple with the adjacent reverse-Hoogsteen A.U pair. The structure of the less highly conserved portion of the loop is less well-defined by the NMR data. A single nucleotide deletion mutant has a very different, open conformation without mismatched base pairs [Varani, G., Wimberly, B., & Tinoco, I. Jr. (1989) Biochemistry 28, 7760-7772]. The implications of the structure for binding of the transcription factor TFIIIA and the cytotoxin alpha-sarcin are discussed. PMID- 8424939 TI - Structural features of an RNA containing the CUGGGA loop of the human immunodeficiency virus type 1 trans-activation response element. AB - A 19-nucleotide RNA containing the CUGGGA loop sequence corresponding to nucleotides 30-35 of the HIV-1 trans-activation response element (TAR) was synthesized in vitro and analyzed by biochemical methods and one- and two dimensional NMR spectroscopy. Diagnostic RNase cleavage patterns were similar for the loops in the full-length HIV-1 TAR and the 19-nucleotide RNA, indicating that they are similar in structure. NMR data showed that the loop is stabilized by base-stacking interactions. The first loop nucleotide is stacked upon the A helical stem, and the loop uridine is stacked upon this cytosine. On the opposite side of the loop, the third loop guanosine is stacked upon the adenosine, which is stacked upon the stem. No specific Watson-Crick or non-Watson-Crick base pairing across the loop was identified. Unusually short interribose distances indicate a significant distortion of the sugar-phosphate backbone centered at the adenosine. Relatively short NMR relaxation times for protons of the adenosine and its adjacent guanosine, as well as rapidly exchanging imino protons, provide evidence for dynamic processes occurring in the loop. PMID- 8424940 TI - Functional roles of the invariant aspartic acid 55, tyrosine 306, and aspartic acid 309 in glucoamylase from Aspergillus awamori studied by mutagenesis. AB - Three mutants, Asp55-Gly, Tyr306-->Phe, and Asp309-->Asn, of Aspergillus awamori glucoamylase (identical to Aspergillus niger glucoamylase) were constructed to elucidate the roles of two conserved regions within fungal glucoamylases. Kinetic studies indicate that both of these regions are closely associated with activity. The Asp55-->Gly mutation decreases the kcat approximately 200 times toward maltose and isomaltose, while KM values remain similar to the wild-type. This localizes Asp55 to subsite 1 of glucoamylase where it affects catalytic activity, but not ground-state binding. The pKa value of the catalytic general acid, Glu179, is 1 pH unit lower in that mutant compared to wild-type enzyme, confirming the proximity of Asp55 to the site of catalysis. Tyr306-->Phe is highly active, but affects binding in subsite 2. It moreover shows enhanced binding in the fourth subsite, suggesting that the conserved region around residue 306 interacts with Trp120, a critical residue that directs conformational changes stabilizing the transition-state structure. Finally, the Asp309-->Asn mutation decreases the kcat for isomaltose hydrolysis around 200-fold, but only 30-fold for maltose. This specific effect on the hydrolysis of the alpha-1,6 linked substrate locates Asp309 to subsite 2. Substitution of Asp309 influences affinities of distant subsites, especially subsite 4, similar to mutations of other carboxylic acid residues situated near subsites 1 and 2. PMID- 8424941 TI - Role of calcium in the adhesion and fusion of bilayers. AB - The interaction forces and fusion mechanisms of mixed zwitterionic-anionic phospholipid bilayers were measured with the surface forces apparatus. The bilayers were 3:1 mixtures of either dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol (DMPC/DMPG) or dilauroylphosphatidylcholine and dilauroylphosphatidylglycerol (DLPC/DLPG), and experiments were carried out in NaCl solutions with and without CaCl2. In NaCl solutions, the forces between either mixed bilayer system were consistent with the DLVO (Derjaguin-Landau Verwey-Overbeek) theory of repulsive electrostatic and attractive van der Waals forces, and fusion did not occur. At high pH (> 6) and in high (20 mM) NaCl concentrations, a short-range hydration force extending about 13 A was evident, indicative of Na+ binding to the surfaces. In the presence of this large hydration repulsion, the interbilayer adhesion was abolished. When CaCl2 was added to the bathing solutions in the presence or absence of NaCl, the bilayers phase separate into small domains, coinciding with the occurrence of a large, long-range attractive force. Fusion occurred readily between the more fluid domains. The phase separations and fusion events could be directly visualized by observing the shapes of the optical fringes used to measure the surface separation and the change in surface profiles with time. The ease of fusion between mixed bilayers in the presence of calcium correlated closely with the strength of the long-range attractive force. This force is attributed to the additional hydrophobic force between domains or domain boundaries due to the exposure of excess hydrophobic groups resulting from the Ca(2+)-induced condensation of the PG- headgroups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424942 TI - Transforming growth factor beta 1: NMR signal assignments of the recombinant protein expressed and isotopically enriched using Chinese hamster ovary cells. AB - The transforming growth factor beta s are a homologous family of multifunctional cytokines that regulate cell growth and differentiation. As a prelude to studies of the solution structure and dynamics of TGF-beta 1, we report virtually complete assignment of 1H and 15N resonances for this 25-kDa homodimeric protein. Recombinant TGF-beta 1 was expressed in Chinese hamster ovary cells. The cells were grown either in a completely 15N-enriched medium or in a medium containing selectively 13C, 15N-labeled amino acids to obtain either uniformly or specifically labeled protein, respectively. Two- and three-dimensional heteronuclear edited magnetic resonance spectra of the uniformly 15N-labeled protein and three samples selectively labeled with 13C and 15N yielded assignments for 96% of the backbone amide and C alpha protons and 87% of the side chain protons. To our knowledge, this is the first report of the use of an animal cell expression system to obtain extensive isotopic enrichment in order to sequentially assign a protein. The methodology described herein for the isotopic enrichment and resonance assignments of TGF-beta 1 should be generally applicable to other eukaryotic proteins expressed by animal cells. PMID- 8424943 TI - Transforming growth factor beta 1: secondary structure as determined by heteronuclear magnetic resonance spectroscopy. AB - Virtually complete backbone NMR signal assignments have been reported for transforming growth factor beta 1 (TGF-beta 1) [Archer et al. (1993) Biochemistry (preceding paper in this issue)]. Herein we report the secondary structure of the protein in solution on the basis of these assignments and proton NOE's observed in a variety of 2D and 3D heteronuclear NMR spectra. Regular elements of secondary structure derived from the NOE data consist of (a) three helices spanning residues Y58-H68, F24-G29, and N5-F8 and (b) several pairs of two stranded antiparallel beta-sheets. The longest two-stranded sheet runs from residue L83 to V106 with a type II reverse turn at G93-R94 and a chain twist at residue N103-M104. These elements of regular structure were confirmed by hydrogen exchange, chemical shift, and coupling constant data. With the exception of residues G46-S53, which exhibit relatively few and weak intraresidue NOE's, residues in the rest of the protein adopt an irregular but well-defined structure. All peptide bonds are trans except for a cis peptide bond between Glu35 and Pro36. The structural characteristics observed for TGF-beta 1 in solution generally agree closely with the recently derived crystal structures of TGF-beta 2 [Daopin et al. (1992) Science 257, 369-374; Schlunegger & Grutter (1992) Nature 358, 430-434]. Several noteworthy differences were observed that may be related to function. PMID- 8424944 TI - Detergent interaction with band 3, a model polytopic membrane protein. AB - The interaction of band 3, the 95-kDa anion-exchange protein of the human erythrocyte membrane, with a variety of nonionic detergents was studied. Band 3 dimers (Stokes radius = 76 A) prepared in octaethylene glycol monododecyl ether (C12E8) could be exchanged into a variety of detergents by size-exclusion high performance liquid chromatography (HPLC), with complete removal of C12E8 from band 3 being confirmed using radiolabeled detergent. Critical micellar concentration (cmc) values, determined for all detergents in the buffer used for HPLC analysis, ranged from 0.47 microM to 223 mM. Band 3 was found to aggregate in all detergents below their cmc, and concentrations of detergents 2-200 times the cmc were required to prevent aggregation. For detergents with a low cmc, it was important to ensure that the concentration of detergent micelles minimally equalled the concentration of protein. Hydrodynamic measurements and cross linking studies showed that band 3 remained dimeric in most detergents above their cmc. Furthermore, circular dichroism and inhibitor binding studies supported the view that band 3 can retain its native structure after detergent exchange. Detergents with short alkyl chains (C8) denature band 3, while detergents with longer alkyl chains (C12) maintained the native structure of band 3. The ability to exchange band 3 into a variety of detergents with the maintenance of native structure is an essential prerequisite for crystallization trials. The results obtained in this study of band 3, a model polytopic (multispanning) membrane protein, may be generally applicable to other membrane proteins. PMID- 8424945 TI - Probing the combining site of an anti-carbohydrate antibody by saturation mutagenesis: role of the heavy-chain CDR3 residues. AB - The carbohydrate-binding site in Fab fragments of an antibody specific for Salmonella serogroup B O-polysaccharide has been probed by site-directed mutagenesis using an Escherichia coli expression system. Of the six hypervariable loops, the CDR3 of the heavy chain was selected for exhaustive study because of its significant contribution to binding-site topography. A total of 90 mutants were produced and screened by an affinity electrophoresis/Western blotting method. Those of particular interest were further characterized by enzyme immunoassay, and on this basis seven of the mutant Fabs were selected for thermodynamic characterization by titration microcalorimetry. With regard to residues that hydrogen bond to ligand through backbone interactions, Gly102H could not be substituted, while several side chains could be introduced at Gly100H and Tyr103H with relatively little effect on antigen binding. There was, however, a preference for nonpolar side chains at position 103H. Substitution of His101H with carboxylate and amide side chains gave mutants with binding affinities approaching that of the wild type; complete side-chain removal by mutation to Gly was tolerated with a 10-fold reduction in binding constant. Analysis of binding by titration microcalorimetry revealed some dramatic thermodynamic changes hidden by the similarity of the binding constants. Similar effects were observed with residue changes in an Arg-Asp salt-bridge at the base of the loop. These results indicate that alterations to higher affinity anti carbohydrate antibodies are characterized by an enthalpy-entropy compensation factor which allows for fundamental changes in the nature of the binding interactions but impedes engineering for increases in affinity. PMID- 8424946 TI - G1- and S-phase synthesis of histone H1 subtypes from mouse NIH fibroblasts and rat C6 glioma cells. AB - The rates of synthesis of histone H1 subtypes in synchronized mouse NIH 3T3 fibroblasts were compared with those of rat C6 glioma cells during the G0, G1, and S phases by using a combination of HPLC techniques and conventional gel electrophoresis. In the mouse cell line, all H1 subtypes, H1a-H1e including histone H1(0), were detectable. In the rat cell line, however, no histone H1a was found. H1c and H1e from both cell lines show in the quiescent state a relatively high specific activity comparable with that of H1(0). After release from the G0/G1 block, the synthesis of H1(0) and likewise that of H1c and H1e increase for a short period. All H1 subtypes have their maximum specific activity at the same time after stimulation. The percentage of total H1 specific activity of H1a, H1b, and H1d increases, those of H1c and H1e remain relatively constant, and that of H1(0) decreases while cells cycle from the G0/G1 to the S phase. These findings support our assumption that H1 subtypes could be classified into three groups with common metabolic characteristics: one consists of H1a, H1b, and H1d; another of H1c and H1e; and a third of H1(0) histone. Moreover, the corresponding H1 subtypes from two different species seem to have similar specific activities during the G1 and S phases. PMID- 8424947 TI - Inhibition of tyrosylprotein sulfotransferase by sphingosine and its reversal by acidic phospholipids. AB - Although tyrosylprotein sulfation has been implicated in the processing of several secretory proteins, nothing is known about the regulation of the enzyme responsible for this event. When poly(Glu6, Ala3, Tyr1) (EAY; M(r) 47,000) was employed as sulfate acceptor, the tyrosylprotein sulfotransferase (TPST) from Golgi membranes of submandibular salivary gland was used to study the effect of various lipids on the expression of its activity. The TPST activity in the Golgi membrane was 38 pmol (mg of protein)-1 (30 min)-1. Approximately 90% of the total activity present in Golgi membranes was extracted by NaCl and Triton X-100 treatment. The Km values of solubilized TPST for EAY and 3'-phosphoadenosine 5' phosphosulfate (PAPS) were 0.04 and 0.25 microM, respectively. Among the various lipids tested, sphingosine showed maximum inhibition of TPST activity followed by sphingomyelin and phosphatidylcholine (PC). Of the two sphingosine analogs tested, threosphinganine was as effective as sphingosine in TPST inhibition, while erythrosphinganine had little effect. In contrast, the acidic phospholipids phosphatidylinositol (PI) and phosphatidylserine (PS) and oleic acid showed slight stimulation. Half-maximal inhibition of TPST was obtained at 150 microM sphingosine (6 mol % when expressed as mole percent of sphingosine to total phospholipids plus Triton X-100). The inhibition was competitive with respect to EAY and uncompetitive with respect to PAPS. The inhibition caused by sphingosine could be reversed by PI, PS, and oleic acid but not by PC and sphingomyelin. Sphingosine inhibition of TPST activity was also observed in the enzyme isolated from several other tissues such as liver, lung, heart, and cerebellum.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8424948 TI - Developmental expression of chicken antithrombin III is regulated by increased RNA abundance and intracellular processing. AB - We isolated and sequenced a 432 bp cDNA to cAT-III, that encoded 115 nucleotides of 5' untranslated sequence, a 17 amino acid long signal peptide and residues 1 88 of the mature protein, and used it to prepare a probe for measuring and correlating the developmental changes of steady-state cAT-III mRNA levels with known changes in antigen levels. Densitometric analysis of nuclease protection (n = 2), Northern blot (n = 4), and slot blots (n = 3) of total RNA from chick livers of 16-day-old embryos to 6-day-old chicks showed a 2.6 +/- 0.5-fold increase in steady-state cAT-III mRNA levels. Assay of functional mRNA levels by in vitro translation of poly(A)+ RNA and specific immunoprecipitation of 35S-Met labelled cAT-III was comparable to RNA analysis (16-day-old embryos vs. 10-day old hatchlings). We evaluated whether there were developmental differences in post-translational secretion which may also contribute to the regulation of the circulating level of this protein. Pulse-chase studies of freshly-isolated hepatocytes from 16-day-old embryos and 10-day-old hatchlings maintained in suspension demonstrated a approx. 5.0-5.5-fold increase in cAT-III levels at steady-state secretion. The above findings indicate that changes in circulating cAT-III levels during late embryonic development are primarily due to increased abundance of cAT-III mRNA. In addition, we postulate that post-translational intracellular processing may account for further differences in circulating protein levels. PMID- 8424949 TI - Cloning and characterization of an alpha-amylase gene from Streptomyces lividans. AB - The alpha-amylase gene (amy) of Streptomyces lividans TK24 was cloned in an amylase deficient mutant strain S. lividans M2. The cloned gene contained an open reading frame (ORF) of 2757 nucleotides (919 amino acids) coding for a protein of 100 kDa. Sequencing of the amino terminus of the extracellular alpha-amylase protein revealed the presence of a signal peptide of 33 amino acid residues. The transcriptional initiation site was mapped by the primer extension method with T4 DNA polymerase and was found to be transcribed from an unique promoter. The alpha amylase protein produced by S. lividans was larger than those derived from other origins. It also contained the four common conserved regions characteristic of other alpha-amylase proteins. PMID- 8424950 TI - Induction of peroxisomal acyl-CoA oxidase by 3-thia fatty acid, in hepatoma cells and hepatocytes in culture is modified by dexamethasone and insulin. AB - The effects of tetradecylthioacetic acid (TTA) (50 microM), dexamethasone (0.25 microM) and insulin (0.4 microM) on induction of peroxisomal acyl-CoA oxidase activity and mRNA levels were studied in short term cultures of Morris 7800C1 and MH1C1 hepatoma cells and of rat hepatocytes. Dexamethasone and TTA resulted in parallel increases in the enzyme activity and the steady state mRNA content in the hepatoma cells. Combination of dexamethasone and TTA resulted in a synergistic and parallel stimulation of both the enzyme activity and the mRNA levels up to 11-12-fold and maximal changes were observed after 14 days of treatment. Semiquantitative immunoblot analyses of acyl-CoA oxidase were in concordance with enzyme and mRNA results. Insulin counteracted the inductive effects of dexamethasone and TTA on all parameters. The half-life of the acyl-CoA oxidase mRNA increased after treatment with the 3-thia fatty acid (t1/2 = 10.0 h +/- 0.4) compared to control (t1/2 = 5.9 h +/- 0.3). However, in combination with dexamethasone there was no further increase in the mRNA stability (t1/2 = 8.0 h +/- 0.3). Southern blot analysis did not reveal any changes on the oxidase gene level in any treatment group. TTA alone or in combination with dexamethasone did not affect the expression of either the glucocorticoid receptor or the peroxisomal proliferator acting receptor (PPAR) steady state mRNA levels. In cultured hepatocytes the acyl-CoA oxidase was modified in similar manner by these treatments, but the changes were less marked. We suggest that the changes in peroxisomal acyl-CoA oxidase activity in hepatoma cells are due to a major effect on the level of mRNA, involving both transcriptional effects and message stabilization. PMID- 8424951 TI - NAD(+)-dependent methylenetetrahydrofolate dehydrogenase-cyclohydrolase: detection of the mRNA in normal murine tissues and transcriptional regulation of the gene in cell lines. AB - NAD(+)-dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate cyclohydrolase, a nuclear-encoded mitochondrial bifunctional enzyme, is detectable in extracts of immortalized and transformed cells but not in most adult tissues (Mejia, N.R. and MacKenzie, R.E. (1985) J. Biol. Chem. 260, 14616 14620). Normal tissues contain low levels of the mRNA, with the exception of thymus and especially testis which contain much higher amounts. The protein could not be detected in any of these normal tissues either by Western analysis or by enzyme activity assay. The elevated level of the mRNA in testis is not dependent on active spermatogenesis. Oncogenic transformation of NIH 3T3 fibroblasts by the Ha-ras oncogene did not significantly affect the steady state level of the dehydrogenase-cyclohydrolase mRNA. The gene in quiescent Balb/c 3T3 fibroblasts is induced by mitogens such as serum and phorbol esters and requires de novo transcription. Post-transcriptionally, the mRNA is stabilized by factors in serum such as insulin-like growth factor-1. The intracellular location of the enzyme and its regulation of expression are consistent with its proposed role in mitochondrial biogenesis. PMID- 8424952 TI - Influence of defects on the electrophoretic, thermodynamic and dielectric properties of a 21 base pair DNA in solution. AB - The thermodynamic and dielectric properties of a 21 base pair DNA have been evaluated and compared with those of samples with some defects. In particular, fragments in which the absence of a phosphate group (nick) or of two nucleotides (gap) causes chain interruptions were studied. Measurements of ultraviolet absorption as a function of temperature at different oligomer concentrations and at various ionic strengths were performed. Dielectric spectroscopy at radiofrequencies (1 MHz-1 GHz) was applied on aqueous solutions of the duplexes at 5 degrees C, where the solutes are thermally stable. Dielectric dispersions with 30-40 MHz characteristic frequencies were defined. The results of melting experiments indicate a thermal destabilization of the oligomers containing the defects. Electrophoretic data and the dielectric results show that the conformations of the nicked and control samples are very similar, while the oligomer with a gap is more compact with a different charge distribution at the ends. PMID- 8424953 TI - Purification and characterisation of Bst LVI restriction endonuclease, a thermostable isoschizomer of ClaI from Bacillus stearothermophilus LV. AB - This work describes the purification and biochemical characterization of BstLVI restriction endonuclease, a thermostable isoschizomer of ClaI, from Bacillus stearothermophilus LV. The enzyme was purified by successive DEAE-cellulose, Affi Gel Blue and Heparin-Sepharose CL-6B column chromatography. A molecular weight of 37,000 was determined for Bst LVI by gel filtration. As expected from thermophilic proteins, the enzyme showed a high stability towards heat and also to other known protein-denaturing agents. PMID- 8424954 TI - A yeast gene (BLH1) encodes a polypeptide with high homology to vertebrate bleomycin hydrolase, a family member of thiol proteinases. AB - We have purified bleomycin hydrolase from yeast (molecular mass 55,000 Da). Using protein sequence-derived degenerate oligonucleotide primers and amplification by polymerase chain reaction, the yeast gene BLH1 was isolated and characterized. The deduced amino acid sequence (483 amino acids) exhibits surprisingly high homology to vertebrate bleomycin hydrolase (43% identical residues and 22% conserved exchanges). It contains three blocks of sequences found conserved in other members of the thiol proteinase family and thought to be associated with the catalytic centre. BLH1 is non-essential under all growth conditions tested. However, in the presence of 3.5 mg bleomycin/ml medium wild-type cells have a slight growth advantage compared to blh1 mutant cells. PMID- 8424955 TI - The POU-domain protein Oct-1 is widely expressed in adult rat organs. AB - The cDNA encoding a rat Oct-1 POU-domain was cloned by the reverse transcription polymerase chain reaction method and subsequently Oct-1 mRNA expression was investigated. Our results show that the POU-domain of Oct-1 has been highly conserved during vertebrate evolution and that Oct-1 mRNA is widely expressed in various organs of adult rat. PMID- 8424956 TI - Molecular cloning of cDNA encoding the phenol/aryl form of sulfotransferase (mSTp1) from mouse liver. AB - The cDNA sequence of the mouse liver phenol/aryl form of sulfotransferase (mSTp1) has been determined. The cloned cDNA consists of 1269 base pairs (bp) and contains an 897 nucleotide open reading frame (ORF) beginning at nucleotide 65, which encodes a 298 amino acid polypeptide of 34.7 kDa. Alignment of mSTp1 to other sulfotransferases shows overall identities of 87% to r-STp, 37% to r-STa, 48% to r-STe, 51% to b-STe, and 37% to h-STa, at the deduced amino acid level. PMID- 8424957 TI - Molecular cloning of a mouse extracellular signal regulated kinase (erk-1). AB - A complementary DNA has been cloned and sequenced for the mouse extracellular signal regulated kinase (erk-1). The nucleotide sequence of the 1.7 kb mouse erk 1 cDNA clone (merk-1-34) is 93.5% identical to the published rat DNA sequence (Boulton et al. (1990) Science 249, 64-67). The mouse clone, as with all other reported sequences, did not contain an in-frame translation initiation codon and, therefore, does not appear to be full-length. Attempts to generate cDNA sequence upstream of the 5' end of merk-1-34 using thermostable reverse transcriptase and chemical melting of the mRNA (methylmercury hydroxide) were unsuccessful. PMID- 8424958 TI - Nucleotide sequence of the gene for phenylalanine ammonia-lyase of rice and its deduced amino acid sequence. AB - The whole structure of the gene for phenylalanine ammonia-lyase of rice, designated as GP-28, was determined. The gene included one intron located between nucleotides 557 and 2048, corresponding to the second and third letters, respectively, of the arginine codon at position 136 which is conserved in the genes of other plant species. The open reading frame was capable of encoding a 716 amino acid sequence, which showed 73.3% similarity with that of parsley. In the carboxyl-terminal region, an arginine-rich sequence common to the rice enzyme was found, which showed low similarity with those of dicot plants. PMID- 8424959 TI - Cloning, sequencing and expression of the mouse mammalian achaete-scute homolog 1 (MASH1). AB - We describe the cloning of a full length cDNA encoding the mouse mammalian achaete-scute homolog 1 (mouse MASH1). Using a ribonuclease protection assay to examine expression of this gene in cell lines, postimplantation embryos and adult tissues, expression was detected between days 10.5 and 16.5 of gestation and in adult brain. No expression was detected in other adult tissues or in most of the cell lines examined. However, differentiation of P19 embryonal carcinoma cells into neuronal cell types by exposure to retinoic acid resulted in the induction of MASH1 RNA expression. PMID- 8424960 TI - Nucleotide and deduced amino acid sequence of porcine interleukin 4 cDNA derived from lamina propria lymphocytes. AB - Total RNA was isolated from in vitro activated lamina propria lymphocytes and used to direct the synthesis of cDNA. Interleukin 4 transcripts were then specifically amplified by PCR. Comparison of the nucleotide sequence with its human homologue demonstrates deletion within the coding region of pig interleukin 4 centred around amino acid residue 70 in the mature human protein. PMID- 8424961 TI - Cloning and nucleotide sequence of the gene for dinitrogenase reductase (nifH) from the heterocyst-forming cyanobacterium Anabaena sp. L31. AB - The nucleotide sequence of an 1655 base pair segment from Anabaena sp. L31 containing the 3' half of the nifU gene, the complete sequence of the nifH gene and the 5' end of the nifD gene is presented. nifH is very highly conserved with the same gene from Anabaena sp. PCC 7120 (91% identical at the nucleotide level; 94% identical at the amino acid level) as are nifU and nifD. The intergenic regions are less well conserved. PMID- 8424962 TI - Intravenous drug use, methadone, and AIDS: ask not for whom the bell tolls. PMID- 8424963 TI - Informational sources in substance abuse. PMID- 8424964 TI - Physician certification in addiction medicine 1986-1990: a four-year experience. The Examination Committee of ASAM. PMID- 8424965 TI - Attention deficits in newly abstinent substance abusers: childhood recollections and attention performance in thirty-nine subjects. AB - Attention deficits were studied among 67 consecutive admissions to an adult tertiary care in-patient substance abuse unit. Fifty-four patients completed the testing; of these 15 were excluded due to confounding variables. The majority of the remaining 39 subjects had evidence of current and/or childhood attention dysfunction. There was no correlation with current performance and childhood recollection of dysfunction even when the effect of gender, race and alcohol abuse were taken into account. There are intriguing educational and therapeutic implications of the similarities between cognitive impairments of newly sober substance abusers and adults with persistence of attention deficit-hyperactivity disorder (ADHD). PMID- 8424966 TI - Alcohol use disorders in cognitively impaired patients referred for geriatric assessment. AB - Alcohol Use Disorders are thought to be underdiagnosed in the geriatric population. A retrospective medical record review was performed on 383 patients who presented for outpatient geriatric assessment from 1985-1990. The record review included data on the alcohol consumption history, age, sex, presence of alcoholic beverages in the home, geriatric psychiatry evaluation, and alcohol related diagnoses. Alcohol Use Disorders were recognized as contributing to medical problems in 10% of patients having a mean age of 78 years. All except one patient were found to be cognitively impaired. In addition, 9% of patients consumed alcohol on a regular basis and this consumption was not considered in the diagnosis, despite the presence of cognitive impairment. Twenty-five percent of cognitively impaired patients were consuming alcohol at the time of evaluation. These results indicate that alcohol consumption and AUD are common in cognitively impaired patients presenting for geriatric assessment. Recognition of Alcohol Use Disorders is essential, as chronic alcohol toxicity represents one cause of potentially reversible dementia. Because there are no validated instruments for alcoholism screening in cognitively impaired elderly patients, evaluation should include a past and present consumption history, a search for alcoholic beverages at the home visit, and possible psychiatric referral. PMID- 8424967 TI - Alcohol and risk/sensation seeking: specifying a causal model on high-risk driving. AB - The authors examine the driving patterns of those at high-risk of drinking driving. A specially designed questionnaire collected data from 878 respondents in alcohol treatment and criminal justice facilities. The LISREL program is used to analyze the causal structure underlying driving behaviors and various alcohol and nonalcohol-related factors. Findings indicate that high-risk driving associates with risk/sensation seeking attitudes: A person who is risk/sensation seeking in general tends to drive at high risk in particular. Further, while chronological age is a significant predictor of high-risk driving, early experience with alcohol intensifies risky driving through its impact on risk/sensation seeking attitudes. PMID- 8424968 TI - Introduction to medical immunology. PMID- 8424969 TI - Genetics of immunoglobulins. PMID- 8424970 TI - Antigen-antibody reactions. PMID- 8424971 TI - The complement system. PMID- 8424972 TI - Lymphocyte ontogeny and membrane markers. PMID- 8424973 TI - Cell-mediated immunity. PMID- 8424974 TI - The humoral immune response and its induction by active immunization. PMID- 8424975 TI - Infections and immunity. PMID- 8424976 TI - Diagnostic immunology. PMID- 8424977 TI - Diagnostic evaluation of humoral immunity. PMID- 8424978 TI - Major histocompatibility complex. PMID- 8424979 TI - Diagnostic evaluation of lymphocyte functions and cell-mediated immunity. PMID- 8424980 TI - Diagnostic evaluation of phagocytic function. PMID- 8424981 TI - Hypersensitivity reactions. PMID- 8424982 TI - Immediate hypersensitivity. PMID- 8424983 TI - Immunohematology. PMID- 8424984 TI - Immune complex diseases. PMID- 8424985 TI - Tolerance and autoimmunity. PMID- 8424986 TI - Organ-specific autoimmune diseases. PMID- 8424988 TI - Rheumatoid arthritis. PMID- 8424987 TI - Systemic lupus erythematosus. PMID- 8424989 TI - Immunosuppression and immunomodulation. PMID- 8424990 TI - Transplantation immunology. PMID- 8424991 TI - Tumor immunology. PMID- 8424992 TI - Malignancies of the immune system. PMID- 8424993 TI - Immunodeficiency diseases. PMID- 8424994 TI - Immunoglobulin structure. PMID- 8424995 TI - Tissues and cells involved in the immune response. PMID- 8424996 TI - Biosynthesis, metabolism and biological properties of immunoglobulins. PMID- 8424997 TI - Marfan syndrome: current and future clinical and genetic management of cardiovascular manifestations. PMID- 8424998 TI - Doppler echocardiographic diagnostic advances in aortic dissection using transesophageal and intraoperative epicardial approaches. PMID- 8424999 TI - When and how to include arch repair in patients with acute dissections involving the ascending aorta. AB - The arch should be included in the repair in patients with acute aortic dissection involving the ascending aorta when (1) the intimal tear is in the arch, (2) rupture of the arch has occurred, (3) the outer wall of the false channel in the arch is tenuous, and (4) the inner wall of the false channel is fragmented. A technique has evolved for patients with acute aortic dissection involving the arch that seems to minimize the difficulties and risks of including the arch, when indicated. Its place will best be determined by additional, prospective, and preferably multi-institutional testing. PMID- 8425000 TI - Genes and gene products involved in Marfan syndrome. PMID- 8425001 TI - The continuing dilemma concerning medical versus surgical management of patients with acute type B dissections. PMID- 8425002 TI - Descending thoracic and thoracoabdominal aortic surgery for aneurysm or dissection: how do we minimize the risk of spinal cord injury? PMID- 8425003 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425004 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. AB - Aortic root replacement in the patient with Marfan syndrome is indicated in the presence of aortic root dilatation more than 6 cm, ascending aortic dissection, or 3 to 4+ aortic valve insufficiency. The objective of operation is to prevent catastrophic aortic rupture or dissection and to treat or prevent aortic insufficiency with consequent left ventricular dysfunction. In over 170 patients undergoing aortic root replacement, operative mortality has been 1%. At mean follow-up of 5 years, the survival rate was 75%. The most frequent serious late postoperative complication has been graft endocarditis (4%). The technique described above carries a low operative risk and late complication rate. We believe it is an important surgical arm in the treatment of cardiovascular complications of the Marfan syndrome. PMID- 8425005 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425006 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425007 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425008 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425009 TI - Composite aortic valve replacement and graft replacement of the ascending aorta plus coronary ostial reimplantation: how I do it. PMID- 8425010 TI - When, why, and how should the native aortic valve be preserved in patients with annuloaortic ectasia or Marfan syndrome? PMID- 8425011 TI - When, why, and how should the native aortic valve be preserved in patients with annuloaortic ectasia or Marfan syndrome? PMID- 8425012 TI - Marketing strategies: creating your own promotional tools. PMID- 8425013 TI - By the inch. PMID- 8425014 TI - The care and feeding of expert witnesses. PMID- 8425015 TI - Clinical perspectives in the treatment of HIV disease: the role of didanosine. Introduction. PMID- 8425016 TI - Clinical perspectives in the treatment of HIV disease: the role of didanosine. Chicago, Illinois, 28 September 1991. PMID- 8425017 TI - In vitro antiviral activity of didanosine compared with that of other dideoxynucleoside analogs against laboratory strains and clinical isolates of human immunodeficiency virus. AB - The in vitro activity of didanosine (2',3'-dideoxyinosine, ddI) against the human immunodeficiency virus (HIV) is generally less potent than that of zidovudine (3' azidothymidine, AZT) and zalcitabine (2',3'-dideoxycytidine, ddC), often by an order of magnitude or more. However, in monocyte/macrophage cell cultures, the potency of didanosine is similar to, or greater than, that of zidovudine but still less than that of ddC. The cytotoxicity of didanosine, including cytotoxicity to bone marrow progenitor cells, is low, and endpoints are often not reached. (The concentration inhibiting 50% of the cell growth [IC50] is > 100 microM.) Thus, in spite of lower potency, didanosine has a high antiviral selective index in vitro. By contrast, zidovudine and ddC are more cytotoxic, with IC50 values < 5 microM. Clinically derived HIV isolates with as high as 30 fold decreases in susceptibility to didanosine in vitro have been described. However, in studies designed to assess the frequency of resistance development after prolonged didanosine therapy, more moderate changes (2- to 5-fold) are seen in general. By contrast, isolates with a 100-fold decrease in sensitivity to zidovudine are frequently found in strains from patients who have received prolonged zidovudine therapy. PMID- 8425018 TI - Longitudinal analysis of responses to oral didanosine therapy following zidovudine therapy in advanced infection with human immunodeficiency virus. AB - The effect on immunologic and virological parameters of up to 24 weeks of therapy with didanosine at daily oral doses of < or = 12.5 mg/(kg.d) was studied retrospectively in 69 patients with advanced disease due to human immunodeficiency virus--i.e., AIDS or advanced AIDS-related complex--who had previously been treated with zidovudine. Patients entered the study with a low CD4 cell count (median, 39/microL) and with evidence of an ongoing depression of bone marrow function. Didanosine therapy was associated with a significant increase in CD4 counts and a prolonged decrease in serum levels of p24 antigen relative to baseline. These changes were more pronounced in the population with baseline CD4 counts of > or = 100/microL. A beneficial effect of didanosine therapy on hematologic parameters was observed in these patients, with increases during therapy of hemoglobin levels as well as white blood cell, granulocyte, and platelet counts. These responses were maximal at weeks 16-20. Further investigations are needed to establish the clinical correlates of these observations. PMID- 8425019 TI - Long-term follow-up of didanosine administered orally twice daily to patients with advanced human immunodeficiency virus infection and hematologic intolerance of zidovudine. AB - This phase 1 trial was conducted to evaluate the safety and tolerance of didanosine (ddI) in subjects with AIDS or AIDS-related complex (ARC) who previously had demonstrated hematologic intolerance of zidovudine. Thirty subjects, 21 with AIDS and nine with ARC, were enrolled. Initially, didanosine was administered orally twice daily for a total daily dose of either 750 mg or 1,500 mg. Subsequently, the dosage for those receiving 1,500 mg/d was reduced to a maximum of 750 mg/d (375 mg twice daily) when data from this and other phase 1 studies showed that the dosage of 1,500 mg/d (750 mg twice daily) was associated with an unacceptable risk of developing neuropathy. The subjects were studied for 46 weeks (mean time; range, 7-122 weeks). The dose-limiting toxic effect observed was peripheral neuropathy, which occurred in eight patients. Other significant toxic effects included pancreatitis in three patients and xerostomia in eleven. In general, didanosine was well tolerated from a hematologic standpoint by the majority of patients during prolonged administration. PMID- 8425020 TI - The didanosine Expanded Access Program: safety analysis. AB - The didanosine Expanded Access Program was designed by the Bristol-Myers Squibb Company (Princeton, NJ) to provide didanosine for treatment of patients who could not be enrolled in clinical trials. This program consisted of two protocols: a treatment IND (investigational new drug) protocol for patients intolerant of zidovudine and an open-label protocol for patients whose clinical condition was deteriorating despite continued therapy with zidovudine. Information from the safety data base derived from study of the first 7,806 patients enrolled has shown that the major adverse events associated with didanosine therapy are pancreatitis (which can be life-threatening), peripheral neuropathy, and diarrhea. Pancreatitis was reported for 5% of the patients. Those with a history of pancreatitis were more likely to develop pancreatitis. In contrast to zidovudine, didanosine has been found to be minimally myelosuppressive. In general, hematologic parameters remained stable, especially for patients who entered the program with normal baseline values. The results of this program suggest that patients pretreated with zidovudine for prolonged periods are able to tolerate didanosine well. PMID- 8425021 TI - Nucleoside analogs: similarities and differences. AB - Among the steps in the replication cycle of human immunodeficiency virus (HIV) that are potential targets for antiviral chemotherapy, the viral DNA polymerase or reverse transcriptase has been a target of choice in research on anti-HIV drugs. Nucleoside analogs--the agents studied in relation to this target--have no intrinsic activity against HIV and must be metabolized to their respective 5' triphosphates by means of kinases, nucleotidases, or other activating enzymes present naturally in cells. The presence and activity of the intracellular enzymes necessary for activation of nucleoside analogs are highly dependent on host species, cell type, and stage in the cell cycle. A great advantage of 2',3' dideoxynucleosides is their intracellular pharmacokinetic profile. The multifactorial mechanisms of toxicity associated with this class of drugs likely explain the different spectra of toxicity observed with the various individual nucleoside analogs and exemplify the uniqueness of each compound. Recently, strains of HIV resistant to 3'-azido-3'-deoxythymidine (zidovudine) were isolated from patients who had been treated with this drug; this resistance probably reflects sequential acquisition of amino acid mutations in the HIV reverse transcriptase. PMID- 8425022 TI - Development of a novel audiological clinical material. AB - Existing ear mould technology depends mainly on dental-based materials and techniques which have retarded provision of effective ear mould services world wide. We provided a range of higher alkyl methacrylate polymeric systems suitable for use in both ear impression construction and ear mould fabrication. This novel clinical material (Otana) and the associated technique for its fabrication have several clinical and economic advantages over traditional materials currently used in audiology. PMID- 8425023 TI - Osteoconduction exerted by methylpyrrolidinone chitosan used in dental surgery. AB - Surgical wounds from wisdom tooth avulsions were medicated with freeze-dried methylpyrrolidinone chitosan, a gel-forming resorbable biopolymer obtained from crab chitosan by chemical modification. Methylpyrrolidinone chitosan promoted osteoconduction and the space left after avulsion was filled with newly formed bone tissue, which conferred desirable mechanical and physiological characteristics to the healed would site. Morphological evidence obtained from biopsies confirmed the radiographic data. Methylpyrrolidinone chitosan was progressively depolymerized by lysozyme and was no longer detected 6 months after surgery. None of the 10 patients reported adverse effects over one year of observation. PMID- 8425024 TI - Fate of bioresorbable poly(lactic acid) microbeads implanted in artificial bone defects for cortical bone augmentation in dog mandible. AB - The fate was examined of poly(lactic acid) microbeads implanted in large artificial defects created in cortical bone of dog mandibles. Two poly(lactic acid) polymers--poly(L-lactic acid) (PLA 100) and poly(DL-lactic acid) (PLA 50)- were used to make microbeads by solvent evaporation with poly(vinyl alcohol) as surfactant. Histological observation of non-decalcified mandibular bone showed that no real bone regeneration existed in the experimental bone defects 18 months after PLA 100 microbeads implantation. The same observation was made 6 months after implantation of PLA 50 microbeads. PLA 100 and PLA 50 microbeads appeared unable to induce regeneration of cortical bone defects of dog mandible, in contrast to previous observations in man for PLA 50 large implants. The failure is tentatively assigned to the presence of poly(vinyl alcohol) at the surface of microbeads. PMID- 8425025 TI - Biospecific haemosorbents based on proteinase inhibitor. I. Synthesis and properties. AB - Sorbents for the removal of proteolytic enzymes from biological fluids were synthesized by immobilization of proteinase inhibitor, ovomucoid from duck egg white, in polymeric hydrogel matrix. The immobilization of the protein inhibitor modified by acryloyl chloride occurs during copolymerization of the unsaturated derivative of inhibitor with hydrophilic monomer and crosslinking agent. It was shown that the sorbent obtained possesses a high affinity for proteinases and is haemocompatible. The use of the haemosorbent for eliminating excess activated proteinase from blood is proposed. PMID- 8425026 TI - Study of blood compatible polymers. III. Copolymers of N-benzyl, N-(2 hydroxyethyl) acrylamide and 2-hydroxyethyl methacrylate. AB - Copolymerization of 2-hydroxyethyl methacrylate (HEMA) and N-benzyl, N-(2 hydroxyethyl) acrylamide (BENAAm) was carried out at different mole ratios of the monomers to obtain copolymers of varying composition. BENAAm content of the copolymers varies between 13 and 70%. Investigation of the interaction of rabbit platelets with these polymer surfaces showed that copolymers with higher BENAAm content inhibit the platelet deformation. Human umbilical cord fibroblast cells proliferated very well on the copolymer surfaces. The cell growth rate on polyHEMA was relatively low. Maximum cell growth was observed on the copolymer having 87% HEMA. PMID- 8425027 TI - Behaviour of the bovine pericardium used in cardiac bioprostheses when subjected to a real fatigue assay. AB - The behaviour of bovine pericardium was studied using a fatigue assay. Twenty three samples were assayed, maintaining the preset initial stress and measuring the time it took for the onset of load loss due to permanent deformation. The results indicated a mathematical relationship defined by the expression: log y = 1.3 - 0.211 log t, where y is the fatigue stress (MPa) and t the duration of the assay. The correlation coefficient was 0.948 (P < 0.001). The safety coefficient of the material diminished significantly as the period of time during which it was subjected to fatigue increased. The theoretical durability of the tissue was much greater than the real durability of the prostheses, which is determined by unsolved problems such as calcification and those derived from suture-related cutting. PMID- 8425028 TI - Care of the neonate. AB - Surfactant administration for respiratory distress syndrome continues to make an impact on neonatal care as large controlled trials are published. Although considered safe, synthetic surfactant administration has been associated with a rare complication in the form of pulmonary hemorrhage. Despite this, significant benefits have been shown. With the approval by the FDA of two surfactant preparations, this treatment is now in widespread use. Although the mortality rate from respiratory distress syndrome and the number of ventilator days are generally decreased, surfactant effect on the incidence of bronchopulmonary dysplasia has been disappointing. Studies of steroid administration for bronchopulmonary dysplasia and steroid side effects have been published in the past year. Steroid use has become widespread for this condition, although many details of its administration and side effects have yet to be worked out. A new area of promise is the use of erythropoietin for anemia of prematurity. Natural historic data on the retinopathy of prematurity have added to our understanding of this condition and have raised new questions on its pathogenesis. Review articles and studies in the area of neonatal encephalopathy stress the need for a more accurate definition of asphyxia and discuss possible prenatal causes of this condition. An extensive review of neonatal jaundice and new recommendations for its treatment in healthy term newborns has been published but remains controversial. PMID- 8425029 TI - Maternal/fetal medicine. PMID- 8425030 TI - Gynecologic oncology. PMID- 8425031 TI - Substance abuse in pregnancy. AB - Substance abuse in pregnancy is a major area of interest for clinicians, researchers, and public health officials. Cigarettes have long been associated with poor reproductive outcomes. Smoking increases the relative risk of placenta previa to 2.6 and of sudden infant death syndrome to 2.2 in white infants. The risk-to-benefit ratio of nicotine replacement therapy is discussed. Alcohol use by pregnant women decreased from 32% to 20% between 1985 and 1988. The long-term sequelae of fetal alcohol syndrome in adolescents and adults are presented. Marijuana is probably the most common illicit drug used in pregnancy. Long-term follow-up showed no difference in outcome until 48 months. However, at 48 months, memory and verbal measures were affected by heavy prenatal marijuana use. Self report of illicit drug use is inaccurate. Routine urine screening for cocaine can show marked variations by locality. Analysis of meconium, urine, and infant and maternal hair along with self-report provides the most accurate information on prenatal cocaine use. The increase in cost of caring for cocaine-exposed infants was estimated. Newborns reveal decreased habituation and increased neonatal stress measures. Long-term sequelae of prenatal cocaine use indicated that smaller head circumference persist and that head size is associated with Bayley mental developmental indexes. However, the Bayley scale measured no difference among the cocaine-exposed, alcohol-exposed, or control groups. PMID- 8425032 TI - Maternal disease and injury in pregnancy. AB - Progress in the management of medical disorders of pregnancy has occurred in many areas. Only salient features are reviewed. The pathophysiology of maternal hyperglycemia in diabetes and its effects on the fetus are explored. Antiphospholipid antibodies, implicated in adverse pregnancy sequelae, come under closer scrutiny in terms of management and correlation with outcome. Studies defining a need for a strict diet for optimal neonatal outcome are presented regarding maternal phenylketonuria. Coagulopathies including protein C deficiency and deep venous thrombosis are reviewed for their impact on pregnancy. Uncommon disorders including cerebrovascular accidents, ureteric obstruction, and myocardial infarction are discussed in relation to management and outcome in pregnancy. PMID- 8425033 TI - Preterm labor, premature rupture of membranes, and cervical incompetence. AB - Prematurity continues to account for 75% of perinatal morbidity and mortality in the United States. The search for accurate clinical methods to predict prematurity and the critical evaluation of treatment strategies for premature labor dominate the recent medical literature. Specific clinical issues addressed during the literature review period include critical evaluation of home tocodynamometry, reevaluation and new investigation into tocolytic agents, clinical role of adjunctive antibiotics, and expanding recognition of tocolytic complications. Laboratory investigations into the role of subclinical genital infections as causes of prematurity continue to show promise. Despite these recent scientific advances, the effective prevention and treatment of prematurity continues to elude our clinical efforts. PMID- 8425034 TI - Hypertension in pregnancy. AB - Selected articles on hypertensive diseases in pregnancy from the past year are highlighted. The concept that endothelial cell dysfunction plays an important role in the pathogenesis of preeclampsia has been strengthened. The initial inciting event is still unclear, but inadequate implantation and decreased antioxidant activity may play a role. Magnesium sulfate is still the anticonvulsant of choice for seizure prophylaxis, but the exact mechanism of its action remains controversial. The ideal screening test to identify those patients likely to develop preeclampsia is still lacking, and the side effects and risks of aspirin prophylaxis are not yet known. The results from the National Institutes of Health-sponsored multicenter trial on this subject should be forthcoming. PMID- 8425035 TI - Prenatal diagnosis and fetal therapy. AB - The protective effect of folic acid supplementation has been demonstrated in patients at risk of neural tube defects by a large, randomized double-blind study. The feasibility of second-trimester screening for Down syndrome, based on the combination of maternal and biochemical markers on maternal blood, has also been verified in two large series from both the United Kingdom and the United States. These results represent major advances in the field of prenatal diagnosis, which, in the future, is likely to rely more and more on better selection by noninvasive testing of high-risk patients. The emphasis has moved away from very early invasive testing due to concerns related to the safety of both early chorionic villus sampling and amniocentesis. Evaluation of renal damage in fetuses with obstructive uropathy by analysis of urinary biochemistry may constitute the basis for more efficient selection of cases amenable to antenatal treatment. Twin-to-twin transfusion syndrome remains a challenge for the future. PMID- 8425036 TI - Perinatal mortality and intrauterine growth retardation. AB - Perinatal mortality has decreased over the past 20 years, and the availability of prenatal care is now recognized as a factor in this improvement. Lupus anticoagulant and anticardiolipin antibodies do not appear to be useful screening tools for perinatal loss. Intrauterine growth retardation remains the second leading known cause of fetal death. Additionally, intrauterine growth retardation causes impaired development lasting into childhood, and the most vulnerable group are those born before 33 weeks' gestation. Newer approaches to the diagnosis and establishment of prognosis for fetuses with intrauterine growth retardation are also reviewed. PMID- 8425037 TI - Diagnosis of breast disease and the role of the gynecologist. AB - The American Board of Obstetrics and Gynecology has recognized the special role of the obstetrician/gynecologist in the diagnosis and treatment of breast disease and indicated in 1985 that it would require a knowledge of breast disease in its certification process. It was also recommended that the obstetrician/gynecologist provide adequate information concerning screening and perform simple diagnostic studies including aspiration of cysts, fine-needle aspiration, and appropriate follow-up studies for patients treated for breast cancer. Because open biopsy often becomes part of the treatment for breast cancer, the Board stopped short of recommending that every obstetrician/gynecologist perform this procedure. The American College of Obstetricians and Gynecologists also has increased its efforts to more clearly define the role of the obstetrician/gynecologist in the diagnosis and treatment of breast disease. Questions have been raised concerning the lack of national standards to evaluate training in the diagnosis of breast disease surgery, and the College has convened a number of workshops to address these issues. This review discusses the issues raised both by the Board and the College. Diagnostic studies including simple procedures such as aspiration of cysts, fine-needle aspiration, and open biopsy are discussed. The role of the obstetrician/gynecologist in the screening examination is emphasized. PMID- 8425038 TI - Prevention of venous thromboembolism in gynecologic surgery patients. AB - To reduce the morbidity and mortality of postoperative deep vein thrombosis and pulmonary embolism, effective prophylactic methods must be used. An assessment of the individual patient's risk is essential in deciding the most appropriate method. In general, women over 40 years of age and all women with other risk factors benefit from some form of prophylaxis. For patients with benign gynecologic conditions, low-dose heparin (every 12 hours) and perioperative intermittent pneumatic calf compression have been shown to be of benefit. Patients at higher risk, such as gynecologic oncology patients, should receive more intense prophylaxis with either low-dose heparin (every 8 hours) or prolonged (5 days) intermittent pneumatic calf compression. Of the two methods, the latter has no significant complications and is therefore our method of choice. PMID- 8425039 TI - Adenocarcinoma of the uterine corpus. AB - Carcinoma of the endometrium is the most common malignancy seen in the female pelvic genital organs. Fortunately, survival is high because most patients are diagnosed in the early stage of the disease. As a result of studies identifying important prognostic factors and a significant number of patients with early clinical stage having extrauterine disease, surgical staging has been adopted for this disease entity. Knowing the exact extent of the disease allows more appropriate adjuvant therapy to be applied postoperatively. It is apparent that the patient with surgical stage I cancer needs radiation therapy only rarely, which obviously saves time and expense and reduces complications. Many patients who have been successfully treated for endometrial cancer may and should enjoy the benefits of estrogen replacement therapy without the risk of increased recurrence of their disease. PMID- 8425040 TI - Multimodality therapy in locally advanced cervical cancer. AB - In the past decade, neoadjuvant and concomitant multimodality therapies have been studied in patients with cervical cancer. The past year has witnessed more pilot studies confirming the feasibility of multimodality therapy. It is now time to move to prospective randomized clinical trials to determine the effects on disease-free intervals, to examine survival rates, and to define the ideal chemotherapeutic regimen and relationship to radiation therapy or surgery. PMID- 8425041 TI - Developmental and organ-specific changes in DNA-protein interactions in the tomato rbcS3B and rbcS3C promoter regions. AB - Sites of DNA-protein interaction were mapped in the promoter regions of two of the five genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase (rbcS) in tomato. The two genes, designated rbcS3B and -3C, are actively transcribed in cotyledons of light-grown seedlings and in leaves, but are transcriptionally inactive cotyledons of dark-grown seedlings, in young and mature tomato fruit, and in roots. The combination and order of conserved DNA sequence elements in the promoter regions of the two genes are essentially identical, but differ considerably from that found in the promoters of the other three tomato rbcS genes, which show different transcription patterns. Nuclear extracts from cotyledons of 7-day-old tomato seedlings, and from leaves and young tomato fruit of mature plants defined multiple DNase I-protected sites in the promoter regions of both genes. The protection patterns were organ-specific, and encompassed previously identified conserved DNA sequence motifs as well as uncharacterized sequences. In contrast, nuclear extracts from mature tomato fruit and roots of 7-day-old seedlings failed to protect any of the promoter sequences, implying that DNA-binding proteins required for transcription of rbcS3B and -3C are inactive in these organs. These results are somewhat surprising since DNA binding proteins from cotyledons of dark-grown seedlings and young fruit interact with the two promoters, although rbcS3B and -3C are not transcribed in these organs. The basis for transcriptional regulation of these two genes is discussed and the detailed pattern of DNase I protection in the promoter regions of the two genes is presented. PMID- 8425042 TI - A 61 bp enhancer element of the tobacco beta-1,3-glucanase B gene interacts with one or more regulated nuclear proteins. AB - We show that a 61 bp fragment derived from the promoter region of the tobacco class I beta-1,3-glucanase GLB gene enhances transcription in Nicotiana plumbaginifolia protoplasts independent of orientation relative to the start of transcription. This fragment leads to a cooperative stimulation of transcription when combined with the cauliflower mosaic virus 35S as-1 enhancer element. The GLB enhancer contains two copies of the sequence AGCCGCC, which is conserved in several genes showing expression patterns similar to the GLB gene, as well as a sequence identical at 6 of 7 bp. Point mutations in these three sequences eliminate the enhancer activity of the 61 bp fragment. Nuclear extracts prepared from leaves of tobacco plants contain one or more putative transcription factors that interact specifically with the GLB enhancer. This factor was much less abundant in nuclear extracts prepared from upper leaves of untreated tobacco plants than in nuclear extracts prepared from upper leaves of ethylene-treated plants or from lower leaves. Since beta-1,3-glucanase genes are expressed at very low levels in upper leaves of tobacco plants, at higher levels in lower leaves, and are induced in all leaves after treatment of plants with the stress hormone ethylene, we conclude that the enhancer element interacts with one or more transcription factors whose binding activity is correlated with gene expression in vivo. PMID- 8425043 TI - Differential expression of U5snRNA gene variants in maize (Zea mays) protoplasts. AB - The small nuclear ribonucleoprotein particles U1, U2, U4/U6 and U5 participate in the removal of introns from pre-messenger RNAs in the nucleus. Three genes encoding U5snRNAs, the RNA moiety of U5snRNPs, have been isolated from maize. As in other plant UsnRNA gene families the three maize U5snRNA genes exhibit sequence variation. Two of the gene variants (MzU5.1 and MzU5.2) are clearly expressed after transfection into maize leaf protoplasts while the third gene variant (MzU5.3) is expressed at very low levels. These different levels of expression cannot be directly correlated with sequence changes in the highly conserved Upstream Sequence Element (USE) required for expression of Arabidopsis UsnRNA genes nor with differential stability of the U5snRNA transcripts. Further sequence elements may therefore have a role in regulating maize UsnRNA gene expression. PMID- 8425044 TI - Variation of proline rich cell wall proteins in soybean lines with anthocyanin mutations. AB - The I locus controls inhibition of anthocyanin accumulation in the epidermal cells of the soybean seed coat and affects abundance of PRP1, a proline-rich cell wall protein in the seed coat. Saline-soluble PRP1 is abundant in the developing seed coats of cultivar Richland (homozygous I, yellow), while it is significantly decreased in the pigmented isogenic mutant T157 (homozygous i, imperfect black). In this report, we examined soluble PRP1 in several cultivars containing alleles of the I locus which affect spatial distribution of pigmentation in the seed coat. We also characterized PRP1 in isolines with allelic variants of several other loci involved in seed coat pigmentation, including T and Im. The T gene is pleiotropic and affects both pubesence color and seed coat pigmentation and structure. Soluble PRP1 was abundant in the developing seed coats of lines with yellow seed (I or ii alleles) regardless of pubescence color, just as in Richland. Likewise, soluble PRP1 was decreased in pigmented seed coats (ik or i alleles) with grey (t) pubescence, as in T157. However, the total seed coat proteins were not extractable from pigmented seed coats with tawny pubescence (i, T genotypes) because they have proanthocyanidins that exhibit tannin properties. The dominant Im allele inhibits seed coat mottling (irregular patches of pigmentation) that occurs if plants are infected with soybean mosaic virus. PRP1 was 35 kDa in mottled (im) isolines and 34 kDa in non-mottled (Im) isolines. PRP2, which is expressed later in seed coat development and in the hypocotyl hooks of soybean seedlings, was also smaller in Im isolines. In summary, some of the anthocyanin mutations affect the quantity of soluble PRP1 polypeptides. while others correlate with structural changes in developmentally regulated proline rich proteins. PMID- 8425045 TI - Genetic inactivation of the psaB gene in Synechocystis sp. PCC 6803 disrupts assembly of photosystem I. AB - The reaction center of photosystem (PS) I is comprised of a heterodimer of homologous polypeptides, PsaA and PsaB. In order to investigate the biogenesis of PS I, the psaB gene was inactivated by targeted mutagenesis in the unicellular cyanobacterium Synechocystis 6803. This mutation resulted in disruption of stable PS I assembly, but PSI II assembled normally. Expression of the psaA gene was not affected by the mutation, but PsaA protein was not detected, indicating that stable PsaA homodimers did not form. The ability to inactivate psaB makes it a viable target for site-directed mutagenesis. PMID- 8425046 TI - Rapid identification of Triticeae genotypes from single seeds using the polymerase chain reaction. AB - An easy and quick protocol has been developed for DNA analysis via PCR. Single cereal endosperm or small leaf pieces can be separately processed in several PCR reactions. The resultant PCR patterns are equivalent to those obtained with standard DNA extraction protocols using either specific or random primers. Intra- and inter-specific variability can be detected. This method allows the analysis of a large number of individuals in early stages prior to the plant sowing. PMID- 8425047 TI - A beta-ketoacyl-acyl carrier protein synthase III gene (fabH) is encoded on the chloroplast genome of the red alga Porphyra umbilicalis. AB - DNA sequencing of a region of the chloroplast genome of the red alga Porphyra umbilicalis revealed an open reading frame of 326 amino acids. Databank searches indicated that this ORF is 34% identical to an E. coli gene (fabH) encoding beta ketoacyl-carrier protein synthase III. In addition, a leucine tRNA gene (trnL(GAG)) was detected just downstream. Neither of these genes are encoded on the chloroplast genomes of land plants. PMID- 8425048 TI - Cytoplasmic male sterility and fertility restoration in wheat are not associated with rearrangements of mitochondrial DNA in the gene regions for cob, coxII, or coxI. AB - In comparing the genetic organization and exploring the molecular basis of cytoplasmic male sterility (CMS) in wheat, mitochondrial DNAs (mtDNA) from Triticum aestivum, T. timopheevi, CMS alloplasmic wheat with T. aestivum nucleus and T. timopheevi mitochondria, and fertility-restored lines were compared by hybridization analysis with specific probes for three gene regions: coxII, cob, and coxI. Minor differences between T. aestivum- and T. timopheevi-derived sources were found for gene regions for coxII and cob. For coxI, there are significant differences between T. timopheevi-derived mtDNAs and T. aestivum mtDNA extending beyond an 8 kb distance. All T. timopheevi-derived mtDNA sources have a chimeric gene region (orf256) with part of the upstream coxI gene region, including some coxI-coding region, preceding coxI. The part of orf256 that does not include any of coxI and the 3'-flanking region of CMS coxI are not found in T. aestivum mtDNA. Neither orf256 nor the CMS 3'-flanking region of coxI are found in T. timopheevi or T. aestivum chloroplastic or nuclear DNA. There do not appear to be DNA sequence differences for the three gene regions studied that are related to either CMS or fertility-restored states. PMID- 8425049 TI - Molecular cloning and characterization of anther-preferential cDNA encoding a putative actin-depolymerizing factor. AB - A cDNA clone, LMP131A, which is preferentially expressed in mature anther was isolated from a lily cDNA library. Northern blot analysis and plaque hybridization experiments showed that the LMP131A mRNA is present at ca. 0.3% of the mRNA in mature pollen and is not detectable in carpel, petal, floral bud, leaf, or root. The clone contains an open reading frame of 139 amino acid residues which shows greater than 40% sequence identity in a 91 amino acid overlap to animal actin-depolymerizing factors (ADF), cofilin and destrin. The sequences at and near the actin-binding site are most conserved. Using the lily clone as a probe, a cDNA clone, BMP1, was isolated from a mature anther library of Brassica napus. The expression pattern of the BMP1 clone was the same as that of the lily clone. The Brassica anther-preferential clone contains an open reading frame which is 79% identical to the lily LMP131A protein. Southern blot analysis showed that there are one or a few copies of the putative ADF genes in B. napus and Arabidopsis thaliana. PMID- 8425050 TI - Organization, expression and nucleotide sequence of the operon encoding R phycoerythrin alpha and beta subunits from the red alga Polysiphonia boldii. AB - The characterization of the operon encoding the alpha and beta subunits of rhodophytan (R)-phycoerythrin (PE) from the macrophytic red alga Polysiphonia boldii is reported. This plastid-encoded operon was cloned, its nucleotide sequence determined, and its expression characterized by northern and primer extension analyses. The arrangement and expression of the PE alpha and beta genes, named rpeA and rpeB, are similar to those of the cyanobacterial (C)-PE genes: rpeB is located 5' of rpeA, with an intergenic region of 64 nucleotides. The two genes are transcribed on a 1.25 kb dicistronic transcript, and each coding region is preceded by a prokaryotic ribosome binding site consensus sequence. Transcription is initiated 95 nucleotides upstream of the initiating methionine codon of rpeB. The promoter region resembles that of prokaryotic genes, with an AT-rich -10 sequence. A direct pentanucleotide repeat (5'-TGTTA 3') was found in the -35 region. This pentanucleotide is present upstream of all PE operons that have been characterized thus far. An extensive inverted repeat is present 3' of rpeA; inverted repeats are found downstream of all PE operons sequenced to date, although the sequence is not conserved. The deduced amino acid sequences from these genes provide complete sequences for an R-PE. Of the amino acid residues 85% are identical to those of bangeophycean (B)-PE from the unicellular red alga Porphyridium cruentum. Conserved residues include cysteines at the bilin attachment sites of C- and B-PEs, aspartates at positions postulated to interact with bilin chromophores, and an apparent consensus sequence for N methylation of an asparagine residue in C-PEs. PMID- 8425051 TI - Developmental and organ-specific changes in DNA-protein interactions in the tomato rbcS1, rbcS2 and rbcS3A promoter regions. AB - DNase I footprinting assays were used to map sites of DNA-protein interaction in the promoter regions of three of the five genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase (rbcS) in tomato. Organ-specific differences in DNase I protection patterns were observed using nuclear extracts derived from cotyledons, leaves, young fruit, mature fruit, and roots of tomato, implying that organ-specific transcription of these genes is controlled at the level of DNA-protein interaction. The three genes, designated rbcS1, -2 and -3A are similarly expressed in cotyledons of dark-grown seedlings, in immature tomato fruit, and in leaves under conditions of water stress. These three genes share at least three DNA sequence motifs, including the G-box sequence, which are apparently not present in the other two tomato rbcS genes. We find protection of one or more of these sequences in the aforementioned organs, indicating that the corresponding DNA-binding proteins could function in directing differential expression of the genes, although functional studies would be required to establish this point. While most of the DNase I protections encompass previously identified conserved sequence motifs and their flanking sequence, we also observe protection of additional sequences, many of which occur in the region of the transcription start site. PMID- 8425052 TI - Genetically engineered resistance against grapevine chrome mosaic nepovirus. AB - Nepoviruses are a group of isometric plant viruses with a genome divided between two-single-stranded, positive-sense, RNA molecules. They are usually transmitted by nematodes and a number of them have significant economic impact, especially in perennial crops such as grapevine and fruit trees. Like all other picorna-like viruses, nepoviruses express their coat protein (CP) as part of a larger polyprotein which is further processed by a virus-encoded protease, a feature which poses specific problems when trying to express the viral coat protein in transgenic plants. A hybrid gene, driving the high-level expression of the CP of grapevine chrome mosaic nepovirus (GCMV) has been constructed and transferred to the genome of tobacco plants. Progeny of CP-expressing transformants show resistance against GCMV. When compared to control plants, fewer inoculated plants become infected and those that become infected accumulate reduced levels of viral RNAs. This protection was also shown to be efficient when plants are inoculated with purified viral RNA. PMID- 8425053 TI - Large unidentified open reading frame in plastid DNA (ORF2280) is expressed in chloroplasts. AB - The chloroplast DNA encodes genes for components of photosynthesis and the transcription-translation machinery; a number of unidentified open reading frames (ORFs) are also present. To determine whether a large ORF in the inverted repeat of chloroplast DNA of tobacco (ORF2280) encodes a chloroplast protein, a conserved region of the ORF was expressed in Escherichia coli. An antibody against the ORF protein was prepared using the purified fusion protein as an antigen. When incubated with proteins from the soluble fraction of tobacco, spinach and Oenothera chloroplasts, the antiserum detects relatively labile polypeptides, which have apparent molecular weights of 170 to 180 kDa. The ORF in tobacco and spinach is large enough to encode a protein of 240-250 kDa, thus it is possible that post-transcriptional or post-translational processing reduces the size of the expression product. Analysis of Oenothera chloroplasts representing four different plastome types revealed endonuclease restriction fragment length polymorphisms in chloroplast DNA indicative of insertion/deletion events in a region of the chloroplast DNA that shared significant sequence similarity with ORF2280. The ORF2280 antiserum was used to demonstrate that there are qualitative differences in the ORF proteins from different Oenothera plastome types. PMID- 8425054 TI - Structure and evolution of a highly repetitive DNA sequence from Brassica napus. AB - A Hind III family of highly repetitive DNA sequences, canrep (canola repeat), was cloned from the nuclear DNA of canola (Brassica napus cv. Westar). The basic units of this family of repeats consists of 176 bp and are arranged in clusters of tandem direct repeats. Each canrep repeat is composed of three related subrepeats of ca. 60 bp. Each subrepeat contains two inverted repeats of about 23 bp and another unrelated sequence of about 12 bp. Based on the internal structure, a possible scheme for the evolution of canrep is proposed. At least two subfamilies of the canrep sequences are present in the genome, as revealed by sequence analyses. In situ hybridization showed that canrep sequences are mainly clustered at centromeric regions of chromosomes. Northern hybridizations indicate that there are no transcripts related to canrep in the total RNAs extracted from plant seedlings. PMID- 8425055 TI - Genes of the R-phycocyanin II locus of marine Synechococcus spp., and comparison of protein-chromophore interactions in phycocyanins differing in bilin composition. AB - R-phycocyanin II (RPCII) is a recently discovered member of the phycocyanin family of photosynthetic light-harvesting proteins. Genes encoding the alpha and beta subunits of RPCII were cloned and sequenced from marine Synechococcus sp. strains WH8020 and WH8103. The deduced amino acid sequences of RPCII were compared to two other types of phycocyanin, C-phycocyanin (CPC) and phycoerythrocyanin (PEC). These three types vary in the composition of their covalently bound bilin prosthetic groups. In terms of amino acid sequence identity RPCII is highly homologous to CPC and PEC, suggesting that the known three-dimensional structures of the latter two are representative of RPCII. Thus the amino acid residues contacting the three bilins of RPCII could be inferred and compared to those in CPC and PEC. Certain residues were identified among the three phycocyanins as possibly correlating with specific bilin isomers. In overall sequence RPCII and CPC are more homologous to one another than either is to PEC. This probably reflects functional homology in the roles of RPCII and CPC in the transfer of light energy to the core of the phycobilisome, a function not attributed to PEC. The genomes of Synechococcus sp. strains WH8020, WH8103 and WH7803 share homologous open reading frames in the vicinity of RPCII genes. The nucleotide sequence extending 3' from RPCII genes in strain WH8020 revealed two open reading frames homologous to components of an alpha CPC phycocyanobilin lyase. These open reading frames may encode a lyase specific for the attachment of phycoerythrobilin to alpha RPCII. PMID- 8425056 TI - Functional analysis of the two homologous psbA gene copies in Synechocystis PCC 6714 and PCC 6803. AB - The cyanobacteria Synechocystis 6803 and 6714 contain three genes (psbA) coding for the D1 protein. This protein is an essential subunit of photosystem II (PSII) and is the target for herbicides. We have used herbicide-resistant mutants to study the role of the two homologous copies of the psbA genes in both strains (the third copy is not expressed). Several herbicide resistance mutations map within the psbAI gene in Synechocystis 6714 (G. Ajlani et al., Plant Mol. Biol. 13 (1989): 469-479). We have looked for mutations in copy II. Results show that in Synechocystis 6714, only psbAI contains herbicide resistance mutations. Relative expression of psbAI and psbAII has been measured by analysing the proportions of resistant and sensitive D1 in the thylakoid membranes of the mutants. In normal growth conditions, 95% resistant D1 and 5% sensitive D1 were found. In high light conditions, expression of psbAII was enhanced, producing 15% sensitive D1. This enhancement is specifically due to high light and not to the decrease of D1 concentration caused by photoinhibition. Copy I of Synechocystis 6714 corresponds to copy 2 of Synechocystis 6803 since it was always psbA2 which was recombined in Synechocystis 6803 transformants. PSII of the transformant strains was found to be 95% resistant to herbicides as in resistant mutants of Synechocystis 6714. PMID- 8425057 TI - The cis-regulatory element CCACGTGG is involved in ABA and water-stress responses of the maize gene rab28. AB - The maize gene rab28 has been identified as ABA-inducible in embryos and vegetative tissues. It is also induced by water stress in young leaves. The proximal promoter region contains the conserved cis-acting element CCACGTGG (ABRE) reported for ABA induction in other plant genes. Transient expression assays in rice protoplasts indicate that a 134 bp fragment (-194 to -60 containing the ABRE) fused to a truncated cauliflower mosaic virus promoter (35S) is sufficient to confer ABA-responsiveness upon the GUS reporter gene. Gel retardation experiments indicate that nuclear proteins from tissues in which the rab28 gene is expressed can interact specifically with this 134 bp DNA fragment. Nuclear protein extracts from embryo and water-stressed leaves generate specific complexes of different electrophoretic mobility which are stable in the presence of detergent and high salt. However, by DMS footprinting the same guanine specific contacts with the ABRE in both the embryo and leaf binding activities were detected. These results indicate that the rab28 promoter sequence CCACGTGG is a functional ABA-responsive element, and suggest that distinct regulatory factors with apparent similar affinity for the ABRE sequence may be involved in the hormone action during embryo development and in vegetative tissues subjected to osmotic stress. PMID- 8425058 TI - Characterization of a spinach gene responsive to low temperature and water stress. AB - The characterization of a cDNA for an 85 kDa spinach protein, CAP85 (cold acclimation protein) that is responsive to cold acclimation and water stress is described. Both transcript and protein levels are increased during cold acclimation and water stress. A novel characteristic of CAP85 is the presence of an 11 amino acid, lysine-rich repeat, common to Group 2 LEAs (late embryogenesis abundant proteins), which is included within a larger repeating motif present in 11 copies. Two other motifs of 8 and 16 residues are also found in three and four copies, respectively. CAP85 like other dehydrins and cold-regulated polypeptides remains soluble upon boiling. Protein blot analyses indicate that CAP85 protein is expressed in all aerial tissues as well as in roots. RNA blots show the presence of mRNA for the 85 kDa protein in leaf, petiole, and root tissue. Cell fractionation studies suggest that CAP85 is predominantly found in the cytosol. PMID- 8425059 TI - The disappearance of an hsc70 species in mung bean seed during germination: purification and characterization of the protein. AB - We have purified a 73 kDa protein from the cytosolic fraction of mung bean seeds. It comprises 0.5-1% of the total protein in seeds. This purified protein is a bona fide hsc70 on the basis of several lines of evidence. First, antibodies against bovine brain hsc70 cross-react with the purified 73 kDa protein. Second, the purified protein comigrates on two-dimensional gels with one of the heat inducible hsc70s in mung bean seedlings. Third, similar to other hsc70 species, the purified 73 kDa protein has a high affinity for ATP. Finally, the hydrolysis of ATP by the purified protein can be stimulated by peptides; ATPase activity increases from 40 nmol/h to 165 nmol/h per mg of protein. The purified mung bean hsc70 autophosphorylates at a substoichiometric level. Moreover, the amount of this hsc70 species diminishes while new species of hsc70s appear after germination, suggesting that the expression of hsc70 in mung bean is subject to developmental regulation. PMID- 8425060 TI - Plant aldolase: cDNA and deduced amino-acid sequences of the chloroplast and cytosol enzyme from spinach. AB - We report the sequences of full-length cDNAs for the nuclear genes encoding the chloroplastic and cytosolic fructose-1,6-bisphosphate aldolase (EC 4.1.2.13) from spinach. A comparison of the deduced amino-acid sequences with one another and with published cytosolic aldolase sequences of other plants revealed that the two enzymes from spinach share only 54% homology on their amino acid level whereas the homology of the cytosolic enzyme of spinach with the known sequences of cytosolic aldolases of maize, rice and Arabidopsis range from 67 to 92%. The sequence of the chloroplastic enzyme includes a stroma-targeting N-terminal transit peptide of 46 amino acid residues for import into the chloroplast. The transit peptide exhibits essential features similar to other chloroplast transit peptides. Southern blot analysis implies that both spinach enzymes are encoded by single genes. PMID- 8425061 TI - DNA amplification fingerprinting of the Azolla-Anabaena symbiosis. AB - The Azolla-Anabaena symbiosis has been used for centuries as a nitrogen biofertilizer in rice paddies. Genetic improvement of the symbiosis has been limited by the difficulty in identifying Azolla-Anabaena accessions and Anabaena azollae strains. The recently developed technique of DNA amplification fingerprinting (DAF) was applied to this problem. DAF uses single, short, oligonucleotide primers of arbitrary sequence to direct amplification of a characteristic set of DNA products by a thermostable DNA polymerase in a thermocycling reaction. The products are separated in polyacrylamide gels and detected by silver staining. DAF could easily distinguish and positively identify accessions of Azolla-Anabaena with DNA extracted from the intact symbiosis. The contribution of prokaryotic Anabaena sequences to the fingerprint of the intact symbiosis, however, ranged from 0 to 77%, depending on the primer sequence. Therefore, DNA extracted from the intact symbiosis would not be suitable for Azolla taxonomy studies. The fingerprints of Anabaena strains isolated by sucrose gradient centrifugation from different species of Azolla could be easily distinguished, and DAF patterns were used to confirm the maternal pattern of transmission of Anabaena in a sexual hybrid. Template DNA extracted from roots was used to produce fingerprints for Azolla without interference from the microsymbiont. Comparison of the patterns from the parents and a hybrid gave strong evidence confirming sexual hybridization. PMID- 8425062 TI - Nucleotide sequence of the potato rDNA intergenic spacer. AB - The large intergenic spacer (IGS) of potato rDNA was sequenced and compared to the IGS sequence of tomato. Both spacers exhibit similar length and architecture. Absence of repeated elements down-stream of the putative transcription initiation site (TIS) in potato is compensated by the larger number of subrepeats upstream of the TIS. Especially high level of similarity (86% and 86.5%, respectively) is found in the AT-rich domain containing the TIS and the region approx. 800 bp upstream of the 18S rRNA gene. PMID- 8425063 TI - Isolation of putative defense-related genes from Arabidopsis thaliana and expression in fungal elicitor-treated cells. AB - Numerous Arabidopsis genes have been cloned that correspond to putative pathogen defense-related genes identified in parsley (Petroselinum crispum). Treatment of Arabidopsis cells with fungal elicitor leads to rapid accumulation of the respective mRNAs with time courses comparable to those observed for their counterparts in parsley. Evolutionary sequence conservation of many of these genes in several plant species suggests they code for important plant functions. PMID- 8425064 TI - Nucleotide sequence and style-specific expression of a novel proline-rich protein gene from Nicotiana alata. AB - cDNA clones encoding a novel proline-rich protein (NaPRP4) have been isolated from a Nicotiana alata stylar cDNA library. The N-terminal part of the derived protein is highly rich in proline (32.2%) and contains several repeats such as Lys-Pro-Pro (7 times) and Pro-Thr-Lys-Pro-Pro-Thr-Tyr-Ser-Pro-Ser-Lys-Pro-Pro (twice); the C-terminal part, on the other hand, has a lower proline content (9.9%) and contains two potential N-glycosylation sites and all the six cysteine residues. Northern blot and in situ hybridisation analyses indicate that expression of the NaPRP4 gene is restricted to cells of the transmitting tract of the style. PMID- 8425065 TI - Structure of the cDNA coding for conglutin gamma, a sulphur-rich protein from Lupinus angustifolius. AB - The sequence of cDNA coding for a sulphur-rich storage protein from Lupinus angustifolius L., conglutin gamma, was determined. The coding region contained an N-terminal leader peptide of 28 amino acids which directly preceded subunits of M(r) 28,239 and 16,517. Extensive sequence homology between the protein encoded by conglutin gamma cDNA and basic 7S globulin from soybean was observed. Sequence homology to proteins from other classes of storage proteins, 11S, 7S and 2S, was limited to short and highly fragmented sequences. The amino acid sequence, Asn Gly-Leu-Glu-Glu-Thr, characteristic of the primary site for post-translational cleavage of the precursors of 11S proteins, was absent from the sequence predicted for prepro-conglutin gamma. It is concluded that conglutin gamma is a representative of a fourth type of storage protein in legumes, distinct from the 11S, 7S and 2S storage protein families. PMID- 8425066 TI - Localization of tRNA genes on the Petunia hybrida 3704 mitochondrial genome. AB - 22 tRNA genes corresponding to 17 tRNA species were localized on the master circle of Petunia hybrida mitochondrial (mt) DNA. Genes for trnN, trnM, trnS-GGA, trnW and trnH are of the 'chloroplast-like' type and presumably originate from promiscuous chloroplast (cp) DNA sequences inserted into the petunia mitochondrial genome. A comparison of the mt tRNAs or tRNA genes population present in two monocotyledonous plants (wheat and maize) and two dicotyledonous plants (petunia and potato) show slight differences in the genetic origin of individual tRNAs. The organization of the petunia mt tRNA genes as well as the number of tRNA gene copies, compared to other plant species, is discussed. PMID- 8425067 TI - Nucleotide sequence and homology comparison of two genes of the sulfate transport operon from the cyanobacterium Synechocystis sp. PCC 6803. AB - The genome of Synechocystis sp. PCC 6803 contains an operon with homology to the sulfate permease of other prokaryotes. We used antibodies raised against cytoplasmic membrane protein to find three genes with strong homology to sbpA, orf81 and cysT genes of the cyanobacterium Synechococcus sp. PCC 7942, Escherichia coli, Salmonella typhimurium and Marchantia polymorpha. It is likely that the permease genes are expressed and the proteins are inserted into the cytoplasmic membrane. PMID- 8425068 TI - [Pulmonary nocardiosis]. AB - The radiological appearances and pulmonary changes during the acute stage and after recovery from pulmonary nocardiosis are described on the basis of 10 confirmed cases. Amongst the 10 patients there were 3 with malignant disease, 1 patient with AIDS, 3 patients following organ transplantation and 3 patients on steroids. In one patient cerebral involvement was demonstrated by MRI. PMID- 8425069 TI - [The computed tomographic differentiation of endobronchial tumors and tumor caused bronchial compression]. AB - The efficiency of computed tomography in differentiating between a bronchial compression due to tumour formation and an endobronchial tumour was tested in comparison to bronchoscopy results. 624 bronchial segments were evaluated in 65 patients with masses in the pulmonary hilus area. Computed tomographical identification of pathological lesions (n = 71) was achieved with a sensitivity of 83% and a specificity of 96%. Out of 52 bronchial segments with endobronchial tumour, 90% showed pathological lesions and 85% were classified correctly. When the morphology of the bronchial lesions (endoluminal mass, smooth or irregularly demarcated bronchostenosis or bronchial displacement) was assessed with CT, only the CT identification of an endoluminal mass could distinguish endobronchial tumour growth with a probability of 91% from bronchial narrowing due to external causes and normal bronchi. PMID- 8425070 TI - [Spiral CT with three-dimensional (3D) surface reconstruction in assessing solitary pulmonary foci]. AB - Recently it has been thought possible to characterise a pulmonary lesion by its vascular involvement. The complex vascular relationships are difficult to demonstrate by the "normal" technique of transverse CT sections. Spiral CT with 3D-surface reconstruction makes it possible to evaluate the vessels accurately and simulates the appearances one would have by looking into the thorax with a demonstration of all vessels lying within the pulmonary space examined by CT. Venous involvement was shown in every one of 18 malignant lesions but was also found in 4 out of 11 benign lesions, arterial involvement occurred in 14 out of 18 malignant and in 2 benign lesions. CT evidence of vascular involvement is in favour of a malignant tumour but is no absolute proof. PMID- 8425071 TI - [CT and MRT of the upper abdomen--is MRT competitive?]. PMID- 8425072 TI - [The 31-phosphorus spectroscopy of space-occupying lesions of the salivary glands. The clinical results and differential diagnosis]. AB - In a prospective study, 15 normals and 20 patients with space-occupying lesions of the salivary glands were examined by MRT images and by in vivo 31-phosphorus spectroscopy. The spectra of malignant tumours showed a significant increase in concentration of phosphomonoesters, phosphodiesters and inorganic phosphates when compared with normals. In addition there was an enormous reduction in creatine phosphates. Increased pH values and marked increase in concentration of inorganic phosphates correlated with poorly vascularised necrotic tumour segments. Concentrations of ATP and PCr were similar to normal muscle tissue. High concentrations of PME and PDE correlated directly with the proliferation of tumour cells and were an important marker for the bioenergy and phospholipid metabolism of the growing tumour. Standardised in vivo 31-phosphorus spectroscopy of space-occupying lesions of the salivary glands provides noninvasive prognostic information on the type and behaviour of the lesion and is complementary to clinical and histological findings. PMID- 8425073 TI - [The sonographic diagnosis of recurrences after parathyroid autotransplantation. The preliminary results of a prospective study]. AB - Reactive renal hyperparathyroidism is a complication of chronic renal failure. The treatment of choice is resection of all four parathyroid glands and autotransplantation of a piece of parathyroid into the muscles of the fore-arm. In a prospective study, the transplant site was examined postoperatively in 35 patients by sonography on at least one occasion. In 3 patients (8.6%) enlargement of the autotransplant was observed and in 2 patients this was associated with clinical and biochemical changes; these were also confirmed histologically. The sonographic appearances of satisfactory and abnormal transplants are described. The value of the method remains to be determined. PMID- 8425075 TI - [The recanalization of thrombosed hemodialysis shunts by oscillating-probe aspiration and with a mesh basket]. AB - Three thrombosed hemodialysis shunts (two polytetrafluorethylene [Goretex] forearm shunts and one Brescia-Cimino shunt) were recanalized by an 8-F oscillational aspiration thrombectomy catheter. The total aspirated blood volume was less than 200 ml in each case. Subsequently, residual wall adherent thrombi were removed by means of a 6-F mesh basket catheter. The mesh basket showed optimal adaptation even to bent vessel or graft segments. The first experience with oscillational aspiration thrombectomy and mesh basket treatment in thrombosed hemodialysis shunts indicate that this technique may serve as a promising alternative to fibrinolysis or surgical shunt declotting. PMID- 8425074 TI - [Local catheter-mediated lysis of the femoral artery: rt-PA versus urokinase]. AB - Percutaneous, catheter-mediated lysis was performed on 56 patients with acute to subacute thrombosis of the superficial femoral artery (Fontaine stage IIb to IV). One half of the patients received rt-PA, the other half urokinase. The clinical parameters of the two groups of patients were largely the same. The therapeutical results were, at about the same treatment costs, better in the rt-PA patients group, in particular with regard to the considerably shorter average treatment period, which lasted on the average for 2 hours in the rt-PA group, vs. 6 hours in the urokinase group. The recanalisation rate was also higher in the rt-PA group (86%) vs. 75% in the urokinase group. PMID- 8425076 TI - [Angiography in infancy and childhood]. AB - Between 1973 and 1991 we performed 160 percutaneous angiograms (130 arteriograms, 30 phlebograms) in children and infants; 12 patients were less than one year and 52 less than ten years old. 44 of the examinations were done by a DSA technique. The examinations were carried out under general anaesthesia except in 8 cases. In 50.7% an arteriogram was carried out for the investigation of a suspected or known tumour, in 9.3% an arteriogram was required following trauma. The most common phlebographic examination was for the demonstration of the spermatic vein; in 27 patients this was done for cryptorchidism or a varicocele. The only complication following a diagnostic angiogram was perforation of one spermatic vein. There were no other complications requiring treatment, such as thromboembolic events. With a total complication rate of 0.6%, angiography in childhood is a relatively safe diagnostic method. PMID- 8425077 TI - [Laser versus rotation angioplasty in the recanalization of chronic femoropopliteal arterial occlusions]. AB - Between June 1987 and July 1989 laser angioplasty, and between July 1989 and December 1991 rotation angioplasty was used as the method of choice for the recanalisation of chronic (minimal duration 3 months) arterial occlusions in the femoro-popliteal region. The technical success rate and final results following supplementary balloon dilatation were identical and there was no significant difference between the two groups (laser 87%, rotation 87.7%). For long occlusions (more than 150 mm), the success rate for rotation angioplasty was 60% and significantly higher than for laser angioplasty at 40%. Complication rates for rotation angioplasty were 24.3%, higher than laser angioplasty with 20.3%. This was due to the higher incidence of emboli of 12.1% compared with the laser technique of 7.3%. Cumulative patency rates after two years showed no significant difference (uncorrelated/correlated: laser 53.4%/67.8%; rotation 56.6%/67.5%). Using lytic and aspiration techniques, the higher incidence of embolisation during rotation angioplasty had no adverse effect on final outcome. If both methods are available, it is advisable to treat long occlusions by rotation angioplasty because of the high immediate success rate, whereas short occlusions are best dealt by laser angioplasty because of the lower incidence of embolisation. PMID- 8425078 TI - [Penile venous outflow occlusion: a comparison of erectile function and cavernosometry before and after percutaneous interventions]. AB - After percutaneous transpenile or retrograde venous occlusion for the treatment of a cavernous leak, the clinical results were correlated with cavernosometric flow measurements in 27 patients. In 15 patients, venous occlusion led to a reduction of the maintenance flow: in patients who showed improvement there was an average reduction of 30 ml/min, in those without improvement or deterioration it averaged 0.3 or 13 ml/min respectively. In 7 patients there was increased flow (up to +21 ml/min). One of these patients showed improved erectile function; the remaining 6 showed no change as was also the case in 5 other patients who had no change in cavernosometric measurements. Our findings suggest a multifactorial cause of the venous leaks. PMID- 8425079 TI - [Somatostatin receptor scintigraphy. A new imaging procedure for the specific demonstration of carcinoids of the small intestine]. AB - Scintigraphy with a radiolabelled somatostatin analog represents a new highly specific approach in the diagnostic work-up of receptor-positive APUD tumours and their metastases. We present our preliminary results with somatostatin receptor scintigraphy in 15 patients with histologically proven midgut carcinoid. 5 out of 6 primary tumour sites (83%) and 90% of the known metastatic lesions could be detected; unknown metastatic lesions were seen in 5 patients. Compared with other nuclear medicine procedures somatostatin receptor scintigraphy is able to detect all tumour sites within hours. This advantage will promote the acceptance of this sensitive and specific imaging modality by the clinicians with regard to preoperative work-up and symptomatic therapy with a somatostatin analog. PMID- 8425080 TI - [Angiography and pressure measurement with a coaxial sliding guidewire: follow-up after renal angioplasty]. PMID- 8425081 TI - [An unusual case of multifocal annular fatty liver]. PMID- 8425082 TI - [The radiological findings in mesenteric lipodystrophy]. PMID- 8425083 TI - [Stenosis of the contralateral iliac artery by a wall stent after the successful recanalization of an obstruction of the A. iliaca communis]. PMID- 8425084 TI - [Intraspinal cartilaginous exostosis as the cause of a Brown-Sequard syndrome]. PMID- 8425085 TI - [Abdominal tumors in children. A comparison between magnetic resonance tomography (MRT) and ultrasonography (US)]. AB - Twenty-one children with predominantly solid tumours detected by US were also examined by MRI (7 neuroblastomas, 2 Wilms tumours, 3 hepatoblastomas, 2 germinal cell tumours, 1 ganglioneuroblastoma, 1 gangliocytoma, 1 Cushing's adenoma, 1 phaeochromocytoma, 1 retroperitoneal rhabdomyosarcoma, 1 diffuse lymphangiectasia of a kidney, 1 splenunculus). The findings from both methods were compared with respect to the identification of the organ involved, extent of the tumour, effect on neighbouring structures and tissue characteristics. US and MRI were of equal value in defining the origin of the lesion and in demonstrating pathological enlargement of lymph nodes. Exact tumour extent could be better demonstrated with MRI because of the ability to perform multiplanar sections and to demonstrate intrathoracic and intraspinal spread. MRI was superior in 9 cases in demonstrating tumour structure and in 6 cases in the evaluation of vascular involvement and vascular anatomy. MRI is therefore recommended as an additional method to US for diagnosis and for treatment planning. PMID- 8425086 TI - Long-term hemodialysis access salvage: problems and challenges for nephrologists and interventional radiologists. PMID- 8425087 TI - Insensitivity of color Doppler flow imaging for detection of acute calf deep venous thrombosis in asymptomatic postoperative patients. AB - PURPOSE: Although color Doppler flow imaging (CDFI) has been shown to accurately depict calf vein thrombosis in symptomatic patients, this technique has not been proved accurate for detection of calf vein thrombosis in a population restricted to asymptomatic postoperative patients. PATIENTS AND METHODS: To evaluate the accuracy of CDFI in asymptomatic postoperative patients, both CDFI and contrast venography were performed on 78 limbs of 76 patients without symptoms of deep venous thrombosis (DVT) who had undergone either hip or knee replacement. CDFI and venographic examination were interpreted blindly with respect to the results of the other modality or clinical findings. Venography was the standard for comparison of results. RESULTS: Fifty-six percent of CDFI examinations of the calf vein were technically adequate. The remaining studies were compromised technically by limb swelling and/or obesity. For the technically adequate CDFI studies, calf vein thrombosis was detected in eight of 10 patients. Calculated sensitivity in this cohort was 80%, and specificity was 97%. The sensitivity of CDFI for acute calf DVT in all patients, regardless of image quality, was 42%. CONCLUSION: These observations suggest that state-of-the-art CDFI is not an accurate examination for acute calf vein DVT in asymptomatic postoperative patients. CDFI is associated with a high rate of technically compromised studies and relatively low sensitivity in studies that are deemed technically satisfactory. These observations do not preclude the use of CDFI in postoperative patients for detection of thrombus extension into the popliteal vein or for detecting thrombosis of more proximal lower extremity veins. PMID- 8425088 TI - Dieulafoy disease: localization with thrombolysis-assisted angiography. PMID- 8425089 TI - Angiographic demonstration of positional stenosis in a failed transobturator bypass graft following surgical thrombectomy. PMID- 8425090 TI - Radiologic placement of long-term central venous catheters: a review. AB - The need for long-term placement of catheters within the central venous system is continually expanding and follows the increasing use of hemodialysis, total parenteral nutrition, and long-term chemotherapy for neoplastic and infectious diseases. Whereas these catheters have traditionally been placed by surgeons in an operating room, it is now clear that they can be effectively placed by interventional radiologists using percutaneous techniques within an interventional/angiographic suite. This review is based on the radiologic percutaneous placement of nearly 1,500 central venous catheters including approximately 500 tunneled Hickman/Leonard catheters, 350 double-lumen cuffed dialysis catheters, and 150 chest wall subcutaneous ports. PMID- 8425091 TI - Infrapopliteal angioplasty: long-term follow-up. AB - PURPOSE: The authors report their experience with 55 intrapopliteal percutaneous transluminal angioplasty (PTA) procedures performed primarily for limb salvage between May 1986 and June 1992. PATIENTS AND METHODS: Forty patients who underwent 55 PTA procedures were followed up prospectively. Multiple risk factors were present in 41 cases (75%). Twenty-three procedures were performed in patients with distal saphenous vein (n = 7) and femoropopliteal (n = 16 [13 were polytetrafluoroethylene]) bypass grafts. Follow-up was available for 54 of the 55 episodes. RESULTS: Technical success was achieved in 52 procedures (95%), with long-term clinical success in 23 of these 52 (44%) at an average follow-up of 25.8 months (range, 1-72 months). Failures tended to occur within the first 3 months. Of the eight PTA procedures in patients with distal saphenous vein bypass grafts, only two remained successes during the course of the study, at an average follow-up of only 2.5 months. Similarly, only two of 15 procedures performed in patients with femoropopliteal grafts remained successes. No complications resulted in emergency surgery or altered the original surgical reconstructive options. CONCLUSION: Infrapopliteal PTA is a safe and effective treatment for native vessel disease in selected patients facing surgical reconstruction for limb salvage but is less durable in patients who have undergone previous bypass procedures. PMID- 8425092 TI - Coaxial catheter-needle system for transjugular portal vein entrance. PMID- 8425093 TI - Microcatheter embolization of non-neurologic traumatic vascular lesions. AB - PURPOSE: The authors report their experience over a 28-month period with embolization of 23 non-neurologic traumatic vascular lesions in 21 patients with use of a coaxial microcatheter coil delivery system. PATIENTS AND METHODS: The injuries included pseudoaneurysms (n = 17), arteriovenous fistulas (n = 3), and sites of extravasation (n = 3) and were caused by gunshot, shotgun, and stab wounds, as well as motor vehicle accidents and iatrogenic trauma. All microcatheter embolizations except one were performed with 2.2-F Tracker-18 catheters inserted coaxially through 5.0-5.5-F guiding catheters. In one case, a coaxial 3-F Teflon catheter was used. In all cases platinum microcoils (almost all non-fibril) and/or straight platinum embolization wires (with fibrils) were used. RESULTS: Twenty-one (91%) of 23 vascular lesions were successfully occluded with use of the microcatheter system. The two cases in which microcatheter embolization failed were successfully managed by using larger catheters and steel coils. Two patients with hepatic vascular lesions (one site of extravasation and a pseudoaneurysm) and one patient with a lower extremity arteriovenous fistula required two procedures each for successful treatment. Procedures were life saving in at least two patients. Two lesions recurred during follow-up ranging from 3 days to 17 months. Both of these recurrences were successfully treated with transcatheter embolization, in one case with use of microcatheters. CONCLUSION: Microcatheter embolization with platinum coils and wires is an effective means for treating traumatic vascular lesions. A coaxial microcatheter system allows for easier, more rapid coil/wire delivery to smaller, spasm-prone arteries in such cases. PMID- 8425094 TI - Young Investigator Award. Percutaneous facet joint fusion: preliminary experience. AB - PURPOSE: The synovium-lined apophysial joint is an important cause of back pain through facet arthrosis and nerve root pressure. The short-term results of two relatively noninvasive treatments--facet block and facet rhizotomy--have been better than their long-term results. The authors propose treatment with fusion of the posterior facet joints in the lumbar spine by means of a simple percutaneous procedure performed under local anesthetic with use of radiologic guidance. MATERIALS AND METHODS: In the first phase of this study the technical feasibility of the concept was established in human cadaver spine. In the second phase a canine model was tested in which bone grafts were surgically inserted into lumbar facet joints. Five male and two female mongrel dogs, weighing 18-25 kg (mean, 20.3 kg), were operated on. Each procedure was done in three steps: First, preoperative computed tomographic (CT) scans were obtained to measure facet depth. Second, surgery was performed under fluoroscopic guidance with use of a threaded guide wire, a small cannulated coaxial drilling system, and hydroxyapatite cancellous bone plugs. Third, early postoperative CT scans were obtained to verify bone plug location. CT was repeated at 4 and 6 months after surgery to check for fusion. The dogs were killed at 6 months, and histologic examination was performed. RESULTS: Successful insertion of the bone plug was achieved in 12 of the 14 facets (86%). One dog died as a complication of the procedure. Fusion occurred in five of the 12 surgically treated facets (42%). CONCLUSION: Drilling and surgical insertion of bone plugs into facet joints of dogs were feasible. The fusion results are promising and may improve with future modifications of the technique. PMID- 8425095 TI - Creation of reentry tears in aortic dissection by means of percutaneous balloon fenestration: gross anatomic and histologic considerations. AB - PURPOSE: To study the safety and efficacy of percutaneous fenestration in aortic dissection, transmural tears in canine and human aortae were created with conventional angioplasty balloons. MATERIALS AND METHODS: Tears created in the aortae of five living dogs were compared with tears created in postmortem specimens. Percutaneous fenestration was performed in a woman with acute type I dissection and ischemic hepatitis who died in multisystem failure, and the balloon tear was documented at autopsy. Additional tears in the human aorta were studied in necropsy specimens of normal, Marfanoid, atherosclerotic, and acutely and chronically dissected thoracic and abdominal aortae. RESULTS: In the canine aorta, transmural balloon tears resulted in rapid death of all five animals, and the tears were approximately 10% longer than tears created post mortem with the same balloon. In human aortic specimens, most transmural and all transseptal tears were linear and were oriented nearly perpendicular to the longitudinal axis of the aorta. Tears that were initiated near calcified plaques or large aortic branches extended in unpredictable directions. The transverse orientation of the tears coincided with the long axis of smooth muscle cells in the media of the intact aorta or the dissection septum. CONCLUSION: Percutaneous balloon fenestration, when performed in areas of the aorta relatively free of atherosclerosis, results in transverse tears in the aortic dissection septum. Percutaneous fenestration of the aortic dissection septum appears feasible and should be considered as a treatment option in carefully selected cases of aortic dissection with ischemic complications. A final conclusion regarding the safety and efficacy of percutaneous fenestration undertaken to relieve organ ischemia requires further clinical experience. PMID- 8425096 TI - Portal decompression after transjugular intrahepatic portosystemic shunt creation with use of a spiral Z stent. AB - PURPOSE: An experimental swine model of acute presinusoidal portal hypertension was used to investigate the feasibility of a spiral Z stent for transjugular intrahepatic portosystemic shunt (TIPS) placement and the correlation between the shunt (stent) size and degree of portal pressure decrease. MATERIALS AND METHODS: Twelve young swine were used. Acute portal hypertension was induced by means of selective injections of absolute alcohol, ethiodized oil, and polyvinyl alcohol sponge particles into intrahepatic portal branches. RESULTS: TIPS was successfully created in all swine by using spiral Z stents that were 6, 8, and 10 mm in diameter; each size stent was deployed in four animals. Being sufficiently flexible, spiral Z stents accommodated for curved shunt tracts. An average of 48% portal pressure decrease was achieved with 6-mm-diameter stents, 61% with 8-mm diameter stents, and 87% with 10-mm-diameter stents. CONCLUSION: These results are in agreement with our clinical experience with use of Gianturco-Rosch Z stents for TIPS formation. PMID- 8425097 TI - Comparison of the mechanical properties of detachable balloons for embolotherapy. AB - PURPOSE: The forces of detachment and the durability of inflation of three different embolization balloons were compared: the Mini-balloon, the detachable silicone balloon, and the gold-valve balloon. MATERIALS AND METHODS: Balloons were considered small (4-6 mm), medium (6-8 mm), or large (8-11 mm), depending on fully inflated diameters. Four Mini-balloons were used in each size category to compare detachment force and inflation duration against three detachable silicone balloons and three gold-valve balloons. RESULTS: All small balloons detached with the same force, while the medium gold-valve balloon and large Mini-balloon detached with less force than their counterparts. The detachable silicone balloons had the shortest release times and a favorable detachment force profile. All balloons remained fully inflated for 11 weeks. CONCLUSION: Most or all vessels should be adequately blocked with any of these devices. PMID- 8425098 TI - Percutaneous management of persistently immature cholecystostomy tracts. PMID- 8425099 TI - Remarks by Louis W. Sullivan, MD, Secretary of Health and Human Services, at the sixth annual meeting of the Association for Academic Minority Physicians, October 23-25, 1992, Washington, DC. PMID- 8425100 TI - Access to medical care and minority populations. PMID- 8425101 TI - Ethnic minorities and access to medical care: where do they stand? AB - Recent research on access to medical care suggests that although minorities may have achieved equity of access during the 1980s, this may no longer be the case. Data collected in the National Medical Expenditure Survey for 1987 are used to examine how African Americans, Asian Americans, and Hispanic Americans fare relative to white Americans on measures of access. Questions on insurance, income, race/ethnicity, place of residence, usual source of care, and use of ambulatory services were asked in a national probability sample of 36,400 US residents. This study found that while only 4.4% of the US population regularly used an outpatient department or emergency room in 1987, 9.9% (P < .0001) of Hispanic Americans, 15.8% (P < .0001) of African Americans regularly used a hospital-based site for their medical needs. Whereas 14% of all Americans were uninsured, 21% (P < .0001) of African Americans and 32% (P < .0001) of Hispanic Americans were uninsured. While 70.6% of all Americans made at least one ambulatory visit to a physician during 1987, 63% (P < .0001) of African Americans, 59% (P < .0001) of Hispanic Americans, and 54.6% (P < .0001) of Asian Americans saw a physician in 1987. Controlling for health status, disparities remained in the use of services for uninsured Americans regardless of race/ethnicity. PMID- 8425102 TI - Immunologic effects of ursodeoxycholic acid in primary biliary cirrhosis. AB - Investigations were undertaken to determine the effect on lymphocyte reactivity of treatment of primary biliary cirrhosis with ursodeoxycholic acid (UDCA). Eight patients with stage 2 and 6 with stage 3 primary biliary cirrhosis were observed. Patients were treated with 300 mg of UDCA three times daily for 1 to 3 years. UDCA reduced jaundice, pruritus, and serum alkaline phosphatase levels. Repeat liver biopsy showed a decrease in inflammation in 3 patients. One patient with stage 3 primary biliary cirrhosis had disease progression despite UDCA treatment. Phytohemagglutinin (PHA) and concanavalin A (con A) were added to mixed lymphocytes from 9 patients receiving UDCA before and after 1 year of treatment. T-cell reactivity was determined by evaluating tritiated thymidine incorporation into DNA. Patients exhibiting an improvement in liver function tests had a significant reduction in lymphocyte reactivity to these mitogens (P < .01). When UDCA was stopped, the response to PHA and con A returned to pretreatment levels, reflecting continued presence of immunologic abnormalities characteristic of primary biliary cirrhosis. UDCA's effect on lymphocyte reactivity appears to be due to a reduction of hydrophobic bile acids, since it does not alter response to mitogens in patients with a normal liver. PMID- 8425103 TI - Rapid development of localized pseudopolyposis in Crohn's colitis. PMID- 8425104 TI - Refolding of recombinant Pneumocystis carinii dihydrofolate reductase and characterization of the enzyme. AB - The isolation of dihydrofolate reductase (DHFR) cDNA sequences from the messenger RNA of Pneumocystis carinii using the polymerase chain reaction is described. The 206-amino acid P. carinii DHFR was expressed to high levels in Escherichia coli inclusion bodies using the T7 promoter expression system. Solubilization of the inclusion bodies in 4 M guanidine hydrochloride and refolding of the recombinant protein in the presence of 0.5% polyethylene glycol 1450 yielded correctly folded DHFR which was purified to homogeneity by methotrexate-Sepharose affinity chromatography. The refolded enzyme was readily crystallized as a ternary complex with NADPH and various inhibitors. The enzyme exhibited a sharp pH optimum with maximum activity at pH 7.0 (turnover number = 6500 min-1). Km values for dihydrofolate (DHF) and NADPH were 2.3 and 3.0 microM, respectively, in 0.1 m imidazole buffer, pH 7. Folate did not act as a substrate. Comparison of the kinetic properties of the refolded enzyme with soluble P. carinii DHFR expressed at low levels in the T7 expression system showed similar pH-activity profiles, Km values for DHF and NADPH, and IC50 values for several known antifolates which were tested as inhibitors of the enzyme. PMID- 8425105 TI - Rapid purification of monomer HIV-2 Tat protein expressed in Escherichia coli. AB - Human immunodeficiency viruses types 1 and 2 encode transactivator proteins, named Tat-1 and Tat-2, that stimulate transcription directed by the viral long terminal repeat sequences. The Tat-1 and Tat-2 proteins are related in protein sequence and mechanism of transactivation. We expressed Tat proteins by in vitro translation and found, as expected, that both Tat-1 and Tat-2 were monomers. Unexpectedly, we found that the Tat-1 and Tat-2 proteins displayed significantly different tertiary structures. As determined by velocity sedimentation and gel filtration, Tat-1 acted as a compact structure and Tat-2 acted as an extended, asymmetric structure. Additionally, we found that the in vitro-expressed Tat-2 protein was significantly less susceptible to aggregation than the Tat-1 protein. This observation led us to overexpress Tat-2 in Escherichia coli and develop a simple and rapid method to purify monomer Tat-2 to approximately 50% purity. These results suggest that large amounts of Tat-2 protein can be purified in suitable form for biochemical and biophysical studies of Tat protein structure. PMID- 8425106 TI - Expression, purification, and functional characterization of the DNA-binding domain of the herpes simplex virus type 1 UL9 protein. AB - The origin-binding protein (UL9 protein) encoded by the herpes simplex virus type 1 (HSV-1) UL9 gene plays a critical role in the initiation of viral DNA replication. The 317-amino acid C-terminal DNA-binding domain (UL9-COOH) of the UL9 protein was expressed as a fusion protein to glutathione S-transferase in bacteria. Milligram quantities of the fusion protein were purified by differential ammonium sulfate precipitation and affinity chromatography over reduced glutathione agarose. The UL9-COOH moiety was cleaved from the fusion protein by proteolysis with thrombin. The purified UL9-COOH displayed specific and stable binding activity toward its DNA recognition sequences within the HSV-1 origin of DNA replication. PMID- 8425107 TI - Resolution and quantification of four metallothionein isoforms from rabbit kidney cells. AB - Reversed-phase high-performance liquid chromatography coupled with an on-line integrating chromatographic data system was used to separate and quantify the four major isometallothioneins MT-1a, MT-2a, MT-2d, and MT-2e present in metallothionein samples from cultured rabbit kidney cells as prepared by gel filtration and/or ion-exchange chromatography. The standard curves for each of these isoforms showed closely comparable linear correlations between the amount of protein applied to the column and their integrated absorbance peak area at 220 nm. The lower limit of quantification is 30 pmol, sufficient to assess basal isometallothionein concentrations and to follow their variation upon metal exposure. PMID- 8425108 TI - A purification method for labile variants of ribonuclease T1. AB - We present a new procedure for the rapid production of ribonuclease T1 variants with decreased stability which could not be purified in satisfying amounts by the existing methods. The major changes from the established procedures are the following. (i) The cells were grown at 28 degrees C rather than at 37 degrees C. (ii) The entire purification was performed at low temperatures (4 degrees C). (iii) Materials for chromatography with high flow rates were used to accelerate protein isolation. (iv) The pH was lowered from 7.5 to 6.0, a condition under which RNase T1 is much more stable. The use of this improved procedure allowed the purification of the labile P39G and P73V variants of RNase T1. By the same technique 300 mg of the wild-type protein could be isolated from 10 liters liquid culture within 3 days. The P39G and the P73V mutations strongly decrease the stability of RNase T1 and the midpoints of the reversible thermal unfolding transition are lowered by 16 and 6 degrees C, respectively, relative to that of the wild-type protein. The decrease in temperature during fermentation and the rapid purification at low temperature and under solvent conditions where the stability of the proteins is high are probably the major reasons for the dramatic increase in yield of these labile variants of RNase T1. Such an approach should be valuable for the production of recombinant proteins in general. PMID- 8425109 TI - Improved procedure for a high-yield recovery of enzymatically active recombinant calf chymosin from Escherichia coli inclusion bodies. AB - The high-yield recovery of enzymatically active recombinant calf chymosin from Escherichia coli inclusion bodies was achieved by optimization of solubilization and renaturation conditions. The solubilization was carried out in 8 M urea at various pHs, at various temperatures, and for various periods of time. The following values were found optimal: 1 h at 31 degrees C, pH 10.4. For successful correct refolding of solubilized prochymosin molecules it was found to be necessary to dilute the solution into an alkaline buffer (pH 10.7) in such a way that the final concentration of urea did not exceed 0.32 M and that of protein 0.275 mg/ml. Our optimized procedure gives about eight times higher yields of enzymatically active chymosin than the current published methods. PMID- 8425110 TI - Baculovirus expression and purification of a soluble, mutant G-protein alpha subunit. AB - The cDNA for the alpha i1 protein that had undergone site-directed mutagenesis to change glycine-2 to alanine was ligated into a baculovirus transfer vector. A recombinant virus was obtained by transfecting Sf9 cells with both the wild-type baculovirus DNA and the transfer vector and screening for recombinant plaques. Infection with the recombinant virus led to a high level of expression of the mutated alpha i1 protein in the soluble fraction of the cell. The protein was purified by ammonium sulfate precipitation, gel filtration, and immobilized dye chromatography. The typical yield was 7.5 mg from two 800-ml cultures. The protein showed immunoreactivity to three different alpha i-specific antibodies. It bound guanosine 5'-(gamma-thio)triphosphate (GTP gamma S) with a stoichiometry of 0.63 to 0.91 mol/mol and with a rate constant (kGTP gamma S) of binding of 0.126 min-1. When GTP gamma S bound, the protein was protected from complete tryptic cleavage. The recombinant protein was able to undergo pertussis toxin catalyzed ADP ribosylation and bind beta gamma subunits but with a reduced affinity compared to that of alpha transducin. Thus using a recombinant baculovirus, a nonmyristylated G protein alpha subunit was abundantly expressed in Sf9 cells and milligram quantities of a functional protein were easily purified. PMID- 8425111 TI - Expression and purification of the adenovirus proteinase polypeptide and of a synthetic proteinase substrate. AB - Simple methods are described for expressing the endoproteinase polypeptides of adenovirus serotypes 2 and 12 in Escherichia coli and for purifying these products from crude bacterial extracts using immobilized metal affinity chromatography. A plasmid for expressing an artificial adenovirus substrate that can be purified by the same method also is described. Cleavage of this substrate by the Ad2 virion proteinase confirmed that the cleavage specificity of the adenovirus proteinase is determined by the four amino acids immediately before the cleavage site. The purified recombinant Ad2 endoproteinase alone was incapable of cleaving the artificial substrate, but cleavage occurred if the artificial substrate was incubated with both recombinant endoproteinase and H2ts1 virions or heat-inactivated wild-type Ad2 virions. These results indicate that, in addition to the 23-kDa proteinase polypeptide, cofactors present in Ad2 virions are required to produce active adenovirus proteinase. PMID- 8425112 TI - The nonbiotinylated form of the 1.3 s subunit of transcarboxylase binds to avidin (monomeric)-agarose: purification and separation from the biotinylated 1.3 S subunit. AB - Avidin-biotin technology is used routinely to purify biotin-containing carboxylases and also proteins that have been chemically coupled to biotin. The 1.3 S subunit of transcarboxylase (TC) studied here is the biotin-containing subunit of TC which not only acts as a carboxyl carrier between the CoA ester sites on the central 12 S subunit of TC and keto acid sites on the outer 5 S subunit of TC but also links the 12 S and 5 S subunits together to form a 26 S multisubunit TC complex. The 1.3 S subunit has been cloned, sequenced, and expressed in Escherichia coli. A method for purifying recombinant 1.3 S subunits from E. coli using avidin (monomeric)-agarose column chromatography has been developed. This affinity-purified 1.3 S was found to be homogeneous by SDS-PAGE, amino acid composition, and N-terminal sequence analysis but had a biotin content of only 28% based on moles of biotin per mole of 1.3 S. This lack of stoichiometry was found to be due to copurification of apo-1.3 S as evidenced by the holocarboxylase synthetase reaction. A procedure for separating the apo- and biotinylated 1.3 S forms using hydrophobic interaction chromatography on an Ether 5 PW column is described. The method is based on the difference in hydrophobicity between apo and biotinylated 1.3 S forms. The copurification of apo and biotinylated forms of 1.3 S on the avidin (monomeric)-agarose column was found to be due to specific interaction with avidin rather than to interaction between apo and biotinylated 1.3 S forms as demonstrated by the fluorescence quenching studies. The results suggest that the avidin-biotin system by itself may not be sufficient to obtain homogeneous biotinyl proteins as nonbiotinyl protein can also bind avidly to such columns. PMID- 8425113 TI - Outcome evaluation and prediction in a comprehensive-integrated post-acute outpatient brain injury rehabilitation programme. AB - Outcomes for 29 individuals with brain injuries (BI) were examined following a comprehensive-integrated rehabilitation programme. From programme admission to completion, proportion living with no supervision increased from 59% to 93%; proportion in transitional or competitive work placements increased from 7% to 59%; unemployment decreased from 76% to 31%. Significant changes in behaviour and functional abilities and achievement of individual goals were also documented with the Portland Adaptability Inventory (PAI) and Goal Attainment Scaling. One year follow-up of 21 graduates indicated general maintenance of gains in independent living and work. At follow-up 86% were living without supervision; 48% were in competitive work; one in transitional work; and 29% unemployed. Although individuals entering treatment less than 1 year after injury showed greater gains than those injured more than 1 year prior to admission, both early and late intervention groups showed significant changes on outcome measures. More extensive disabilities as measured by the PAI had a negative impact on programme outcome. Except for reading ability, neuropsychological measures obtained prior to admission did not significantly predict outcome. Programme costs are reported. Results indicate that the group-oriented comprehensive-integrated approach to post-acute brain injury rehabilitation is effective and cost-effective, and recommend early intervention for optimal outcomes. PMID- 8425114 TI - Critical factors associated with the successful supported employment placement of patients with severe traumatic brain injury. AB - A number of investigations have recently demonstrated the effectiveness of supported employment programmes in improving the vocational status of clients with traumatic brain injuries. The present study investigated a sample of 39 individuals participating in a return to work programme emphasizing a supported employment approach to identify key functional characteristics which differentiated successful and unsuccessful clients. Clinical ratings of employment specialists were used to identify two groups of individuals rated least and most difficult to place and maintain. Results indicated that the two groups differed markedly on key employment outcomes. Individuals rated most difficult tended to be younger, possess functional limitations such as visual and fine motor impairments, and display significant deficits in numerous work-related skills. Recommendations are made for applying these findings to assessment, placement, and training activities within vocational rehabilitation programmes for persons with traumatic brain injuries. PMID- 8425115 TI - The adversarial alliance: developing therapeutic relationships between families and the team in brain injury rehabilitation. AB - Families of individuals who have survived brain injuries experience significant distress, and may resist accepting their relative's neurobehavioural deficits. Staff who work with brain-injured patients and their relatives are charged with the seemingly paradoxical task of helping families support rehabilitative efforts and be goal-oriented, while simultaneously communicating often negative realities about prognosis. In the midst of what may be an intermittently conflict-laden relationship, families and staff must become synergistically involved in a therapeutic partnership. This paper defines aspects of this 'adversarial alliance' which is often established between families and staff. The relationship between patient discharge outcome and perceived family stress and satisfaction with the rehabilitation programme was reviewed. Data analyses yielded the following conclusions: families evaluated retrospectively to have been 'highly stressed' were also perceived to experience more conflict with the rehabilitation team; family stress was related to poorer adjustment to the patient's disability (at admission); greater family/team conflict correlated with lower cognitive and physical functioning at admission, longer length of stay, younger patient age, and lower programme satisfaction. Implications for programme development and treatment guidelines are discussed. PMID- 8425116 TI - Use of the Recognition Memory Test in traumatic brain injury: preliminary findings. AB - The suitability of Warrington's Recognition Memory Test (RMT) for detecting memory impairment after traumatic brain injury was investigated in a pilot study of 36 patients with moderate and severe brain injuries. The patients showed significant memory deficits on both RMT subtests, Words and Faces, with greater impairment on the Faces subtest. No significant gender differences were found. The left hemisphere lesion group obtained the lowest mean score on the Words subtest, and the right hemisphere lesion group had the lowest mean score on the Faces subtest, although these differences did not reach statistical significance. At the 90-94% specificity level the sensitivity of the Faces and Words subtests for the detection of memory impairment after traumatic brain injury was 81% and 63%, respectively. The RMT appears to be a promising instrument for measuring and describing patterns of memory impairment after brain injury. PMID- 8425117 TI - Hypernasality in dysarthric speakers following severe closed head injury: a perceptual and instrumental analysis. AB - Hypernasality in the dysarthric speech of 20 severely closed-head-injured (CHI) subjects was investigated using both perceptual and instrumental techniques. A perceptual analysis of the speech of the CHI subjects was performed using a four point rating scale for hypernasality. Instrumental assessment was carried out using a computerized accelerometric technique yielding a nasal coupling index. Results revealed a high incidence of perceived hypernasality (95%) in the speech of subjects in the CHI group. More than half of these subjects exhibited hypernasality of speech to a moderate to severe degree. When compared with a control group matched for age and sex the severely CHI subjects were perceived as being significantly more hypernasal. Instrumental assessment revealed that the functioning of the velopharyngeal valve in the group of CHI subjects was significantly impaired compared to the control group. The study highlighted the need to evaluate the perceptual and instrumental assessment results for the severely CHI subjects on an individual basis. The clinical implications of these findings are discussed. PMID- 8425118 TI - Evaluation of an innovative cost-effective programme for brain injury patients: response to a need for flexible treatment planning. AB - There is clinical support for the hypothesis that brain-injured patients benefit from the high degree of structure provided by an inpatient rehabilitation programme. However, they often experience mild to moderate regression in behavioural and functional skills when they are discharged to home. This paper presents an evaluation of an innovative phase within an acute brain injury programme designed to facilitate the patient's approach towards independence. 'Step-Up' programme patients, who spent the last weeks of their inpatient length of stay residing in a transitional living setting, were compared with inpatients who participated in the regular therapy programme. Eighteen months of descriptive data regarding cognitive (Rancho) and functional (Barthel) levels for 31 patients are compared. Based on survey data, the Step-Up patients reported more independence in activities of daily living than did inpatients. The Step-Up programme also proved to be more cost effective than the inpatient alternative. Challenges to innovative programming presented by payors, families, and clinicians are discussed. PMID- 8425119 TI - Chlorpromazine-induced psychosis after brain injury. AB - Antipsychotic agents, most often used for treatment of schizophrenia, are sometimes prescribed for the agitated patient with an organic brain disorder. We report the case of a brain-injured patient who was prescribed chlorpromazine for agitation and who developed a delusional state while taking this antipsychotic agent. The emergence of this delusional state coincided with the exacerbation of certain cognitive deficits. Possible mechanisms for this phenomenon are discussed. Caution is advised when prescribing neuroleptics for patients with traumatic brain injury, especially those agents with significant cognitive side effects or with a significant potential to precipitate seizures. PMID- 8425120 TI - Methods of increasing client acceptance of a memory book. AB - This case history describes the successful use of a commercially available memory book by a brain injury survivor, including a description of the characteristics which made the book acceptable. Implications of the normalization principle for memory rehabilitation are also discussed. PMID- 8425121 TI - Computer hardware and operating systems. PMID- 8425122 TI - Cocaine in the UK--1991. AB - More than 100 years after Freud's original endorsement of the drug, the use of cocaine is a problem for both users and for society, which struggles to organise effective responses to the epidemic of the last decade. During the 1980s the rapid spread of smokeable cocaine (including 'crack') was seen in the Americas (particularly the US). The initial simple predictions of an identical European epidemic were mistaken. The available data on the extent of cocaine use and of cocaine problems in the UK are examined. New forms of cocaine have been developed by black-market entrepreneurs ('freebase' and 'crack'), and new technologies have emerged for their use; with these new technologies have come new effects and new problems. The general psychiatrist now needs a knowledge of directly and indirectly related psychopathology which has an increasing relevance to the diagnosis and management of the younger patient. PMID- 8425123 TI - The psychological adjustment of the Chinese community in Britain. A study of two generations. AB - The psychological health and adjustment to life in Britain of a sample of first- and second-generation Chinese immigrants were measured. It was predicted that problems with the English language, inadequate social support, value differences, and unfulfilled expectations would induce more symptoms of psychological distress and depression in first-generation than in second-generation Chinese immigrants. Overall psychological health, and hence adjustment, was good. There was evidence for language problems and unfulfilled expectations, but not social support and value differences, being linked to mental health in the second generation. Evidence linking mental health to other personal variables was found in both generations. PMID- 8425124 TI - The psychopathology of Gilles de la Tourette's syndrome. A controlled study. AB - The present study compares adults with Gilles de la Tourette's syndrome (GTS) with depressed adults and normal controls on questionnaires measuring obsessionality, depression, and anxiety. The GTS and depressed groups scored significantly higher than the normal controls on all measures. The GTS subjects had similar scores on measures of obsessionality to those of the depressed subjects, but significantly lower scores on measures of depression and anxiety. This suggests that obsessionality is a prominent feature of GTS, and that the psychopathological profile is different to that of patients with major depressive disorder. PMID- 8425125 TI - Autobiographical memory in depression: state or trait marker? AB - Two cognitive measures were used to assess 22 patients who met DSM-III-R criteria for major depressive disorder: the Autobiographical Memory (AM) test and the Dysfunctional Attitude Scale. They were followed up over seven months. Measurement of dysfunctional attitudes did not predict outcome at seven months. Overgeneral recall on the AM test at initial assessment, especially for emotionally positive memories, was highly correlated with failure to recover from depression and accounted for 33% of the variance in HRSD score at follow-up. Overgeneral recall of emotional memories did not change during follow-up. It is suggested that overgenerality is a trait marker indicating vulnerability to persistent depression. PMID- 8425126 TI - Drug treatment of personality disorders. PMID- 8425127 TI - Suicide and Asian religions. PMID- 8425128 TI - What is the incidence of conversion pseudodementia? PMID- 8425129 TI - Multiple personality. PMID- 8425130 TI - Self-poisoning and general elections. PMID- 8425131 TI - Tattoed female psychiatric patients. PMID- 8425132 TI - Simple schizophrenia--a forgotten diagnosis. PMID- 8425133 TI - Phenomenology and schizophrenia. PMID- 8425134 TI - Nutmeg or mabi bark? PMID- 8425135 TI - Reform of the Mental Health Act 1983. An effective tribunal system. PMID- 8425136 TI - Psychopharmacology and the ethics of resource allocation. AB - The introduction of clozapine to current psychiatric practice is considered against a background of potential problems of resource allocation posed by the development of a number of 'budget-busting' drugs. It would appear that clinicians may increasingly have to operate within a climate in which the rights of individual patients to expensive treatments will seem to be pitted against the abilities of their communities to afford such treatments. Both clinicians and pharmaceutical companies have roles in the development of such conflicts. PMID- 8425137 TI - Assessment of costs and benefits of drug therapy for treatment-resistant schizophrenia in the United Kingdom. AB - An analysis was conducted on the basis of available data to assess the economic consequences of clozapine therapy for people with moderate to severe schizophrenia in long-stay institutions or staffed group homes, with a view to providing an estimate of the likely costs and benefits of the drug. Data from a cost-effectiveness study conducted in the US, supplemented by other literature sources, were used to construct a clinical decision tree for likely clinical outcomes for such patients. A panel of UK psychiatrists provided consensus on how these patients would have been managed in the UK. The costs associated with each patient outcome were estimated, and a sensitivity analysis performed to test the assumptions made. For the patients themselves, clozapine would lead to a net gain of 5.87 years of life with no disability or only mild disability. The base case analysis showed that the direct costs of using clozapine were 91 pounds less per annum (or 1333 pounds per lifetime) than for standard neuroleptic therapy, when the effect on all health-care resources was taken into account. In addition, the sensitivity analysis showed that clozapine would be cost-saving or cost-neutral under many different assumptions. A prospective health economic study with clozapine in the management of schizophrenia would be desirable to confirm these results. PMID- 8425138 TI - The effect of clozapine on cognition and psychiatric symptoms in patients with schizophrenia. AB - Psychiatric symptoms and cognition were assessed in 13 patients with schizophrenia, one patient with schizoaffective disorder, and one patient with psychosis not otherwise specified while they received a conventional neuroleptic and again after an average of 15 months on clozapine. Despite improvements in psychiatric symptoms, attention, memory, and higher-level problem-solving were essentially unchanged. This suggests that certain cognitive deficits are relatively independent of psychotic symptoms in schizophrenia, and are probably central and enduring features of the disorder. Cognitive disability appeared to have been rate-limiting in the sample's rehabilitation, as patients' social and vocational adjustment remained marginal during the study. We also observed that treatment with clozapine was associated with a decline in some memory functions; the potent anticholinergic properties of the drug may have been responsible for this. PMID- 8425139 TI - Psychiatric problems following bereavement by murder or manslaughter. AB - Bereavement by murder or manslaughter is often associated with a high incidence of factors which increase the risk of lasting psychological problems after bereavement. In this study it appears that self-perpetuating vicious circles often accounted for the persistence of symptoms, which fitted the diagnostic categories of post-traumatic stress disorders, anxiety states, panic syndromes, obsessive revenge-seeking, and depression. Therapeutic approaches should be aimed at interrupting these vicious circles and fostering the work of grieving. PMID- 8425140 TI - Psychiatric morbidity in survivors of organised state violence including torture. A retrospective series. AB - The case notes documenting the psychological well-being of 100 survivors of torture and other forms of organised state violence were analysed retrospectively. The most common diagnoses were post-traumatic stress disorder (PTSD), major depression, and somatoform disorders. Of these, PTSD showed the strongest association with experience of torture. It is possible that PTSD has a dimensional nature, and that reactions to different stressors are heterogeneous. PMID- 8425141 TI - Lymphocyte response in depressed patients and subjects anticipating bereavement. AB - Lymphocyte response to stimulation with phytohaemagglutinin (PHA) was assessed in 11 patients with major depression and 8 subjects anticipating bereavement, in order to examine whether altered immune response (PHA stimulation index) was more closely related to depressed mood than to sleep and weight changes. No significant relationship was found between sleep or weight changes and immune response. Depression, measured using the HRSD (with and without sleep scores) and the BDI, was related differently in the two groups. For the depressed patients, increasing depression was associated with reduction in immune response; among those anticipating bereavement (with low depression scores), increasing depression was associated with enhanced immune response. A regression curve using data from both samples demonstrated an inverted 'U'-shaped curve relating immune response to mild and severe depressed states. The results of this study suggest a hypothesis that may explain previous discrepant results and which requires testing on more subjects. PMID- 8425142 TI - Premorbid social underachievement in schizophrenia. Results from the Camberwell Collaborative Psychosis Study. AB - In an investigation of the timing and precursors of social decline in schizophrenia and affective psychosis, 195 subjects from the Camberwell Collaborative Psychosis Study were currently of lower social class than were their fathers. A comparison between father's occupation and proband's best premorbid occupational level indicated underachievement confined to DSM-III schizophrenia, there being no such effect in affective psychosis. Decline in social status following onset of psychosis, analysed by comparing best premorbid occupation with current occupation, was marked in both schizophrenia and affective psychosis, indicating a non-specific effect. Schizophrenic patients who failed to achieve their fathers' social status had poorer educational qualifications than those who equalled or bettered their paternal social class, despite similar premorbid IQ (NART) scores and age at onset of psychosis. These results indicate that schizophrenia may be manifest before the onset of psychosis, and lend weight to the notion of a developmental origin to this disorder. PMID- 8425143 TI - Life events and psychosis. Initial results from the Camberwell Collaborative Psychosis Study. AB - Data from the Camberwell Collaborative Psychosis Study were used to examine the proposition that there is an excess of life events preceding the onset of psychoses of all types. Of 97 patients from the study who had episodes within the past year that were datable, 51 had developed psychotic symptoms from an essentially symptom-free state, 29 had been suffering only from neurotic symptoms, and 17 had experienced a marked exacerbation of psychotic symptoms. DSM III diagnoses were collapsed into three major groups: 51 cases of schizophrenia; 31 cases of mania; and 14 cases of depressive psychosis. Life-event histories were taken for the six months before onset, and when these were compared with equivalent histories from a psychiatrically healthy sample from the local general population, there was a significant excess of life events, particularly in the three months before onset of psychosis. This was apparent in all groups, and remained even when events were restricted to the independent category. The excess of events began rather earlier than has been found in previous studies. In our view, this study provides some of the strongest evidence for a link between life events and the emergence of psychotic symptoms. PMID- 8425144 TI - The influence of age and sex on the onset and early course of schizophrenia. AB - A new standardised interview for the retrospective assessment of onset and early course of schizophrenia (IRAOS) was used to study the influence of age and sex on time of onset and psychopathology before first admission in 267 schizophrenic patients admitted for the first time. Mean age at onset, according to various operationalised definitions, differed by three to four years between the sexes. The age distribution at the earliest sign of mental disorder showed an early and steep increase until the age of 25 in males, and a delayed and smaller increase in females, with a second peak in women aged 45-79. Schizophrenia began with negative symptoms in 70% of cases, appearing two to six years before admission, and all positive symptoms appearing up to two years before. Both positive and negative symptoms accumulated exponentially. The early course of the disease was similar across age groups, except there was a longer period of negative symptoms before first admission in late-onset schizophrenia in women. The few significant age differences in symptoms were presumably due to general age-dependent reaction patterns like anxiety and depression or the cognitive development of personality, as indicated by an increase in fully elaborated positive symptoms, especially systematised paranoid delusions, with age. PMID- 8425145 TI - A twin study of psychosis and criminality. AB - Lifetime criminal and psychiatric histories were examined in a consecutive series of 280 individuals of twin birth with a diagnosis of major functional psychosis who were seen and followed up at the Maudsley Hospital between 1948 and 1988. Their 210 co-twins, 35% of whom had a similar diagnosis, were ascertained and followed up over the same period. In the absence of reliable general-population estimates for lifetime conviction rates, co-twins were used as case controls. Among the 220 complete pairs, significantly more probands (25.7%) than co-twins (14.0%) were convicted, although there was no evidence for an independent genetic basis for criminal behaviour. Criminal conviction was significantly related to psychiatric diagnosis. There were specific patterns of offending, particularly among the schizophrenic men, who were also significantly more often convicted (48.6%) than the men with affective psychosis (19.4%), and more likely to receive a prison sentence. The schizophrenic patients were younger at their first conviction (mean age 22.6 years v. 30.8 years) and they had committed more violent offences than the affective group. In both diagnostic groups, ages at first psychiatric contact and first conviction were highly correlated. PMID- 8425146 TI - The community's tolerance of the mentally ill. AB - A survey of attitudes to mental illness was conducted in a quota sample of about 2000 subjects in Malvern and Bromsgrove. Factor analysis showed three main components - benevolence, authoritarianism, and fear of the mentally ill. Residents of Bromsgrove, which is served by a traditional mental hospital, were slightly more tolerant than those living in Malvern, which has a community-based service, and has seen the closure of two mental hospitals in its vicinity during the last 10 years. The main demographic determinants of tolerance are age, education, occupation, and acquaintance with the mentally ill. PMID- 8425147 TI - A comparison of responses to the mentally ill in two communities. AB - Vignettes representing mentally ill people were presented to about 2000 randomly selected residents in Bromsgrove, served by a mental hospital, and Malvern, served by a community-based psychiatric service. They were asked about the likely cause of the condition, what action they would take, and what agencies were most likely to help. In Malvern, residents seemed more enterprising in involving various agencies and more tolerant. It is possible to derive simple 'action scores' as an indicator of such tolerance. Although there were significant demographic differences between subgroups, overall identification of vignette subjects as mentally ill was surprisingly low, and so was knowledge both of community psychiatric nurses as an agency, and of the location of dispersed treatment facilities in both areas. PMID- 8425148 TI - Hand lipomas: detection and characterization by magnetic resonance imaging. AB - The initial clinical examination of two patients with a mass lesion of the palmar aspect of the hand failed to suggest the cause. Subsequently, the patients were examined with magnetic resonance imaging, which demonstrated that the lesions were lipomas. The scans revealed masses with smooth, lobulated margins and internal septations of low signal intensity; the masses exhibited high signal intensity in T1-weighted spin-echo images and intermediate signal intensity in T2 weighted images, features consistent with a lipoma. PMID- 8425149 TI - Old renal transplants examined by Doppler ultrasonography. AB - Twenty-four ultrasonography (US) studies of 21 renal transplants that had been in place for 1 to 19 (median 3) years were evaluated. The absolute resistive indices (RIs) determined during Doppler US at various arterial levels and the differences in RI between the levels did not correlate with the clinical diagnosis of chronic rejection. This study suggests that measuring RI values with Doppler US in people who received renal grafts as children does not currently contribute additional information for diagnosis or management. PMID- 8425150 TI - Hypertrophic pyloric stenosis in the neonate--diagnostic criteria revisited. AB - The authors analysed 45 ultrasonography (US) studies of 43 infants with surgically confirmed hypertrophic pyloric stenosis (HPS) to ascertain whether the current US criteria for pyloric stenosis are applicable to all infants, including those 30 days of age and younger. Most articles in the radiologic literature cite a muscle thickness of 4 mm or greater and a pyloric canal length of 17 mm or greater as diagnostic of HPS. However, the results of this study suggest that in infants 30 days of age or younger the muscle thickness considered diagnostic for HPS be 3 mm or greater. PMID- 8425151 TI - Quality of mucosal coating in double-contrast barium enema studies: comparison of two barium preparations. AB - To determine which of two types of barium preparation, a dry powder (AC-14, E-Z EM, Montreal) or a premixed suspension (Liquid Polibar Plus, E-Z-EM), gives better mucosal coating for double-contrast enema studies, the authors compared the results of two series of 80 examinations each, one for each of the preparations. The films were retrospectively reviewed by two independent observers, who did not know which preparation had been used for a given examination. Mucosal coating was graded on a five-point scale and the amount of barium on a three-point scale. Each area of the colon--the cecum, the ascending, the transverse and the descending colon and the rectosigmoid--was rated separately. The premixed barium suspension yielded significantly better mucosal coating than the dry powder in all areas of the colon (p < 0.01). There was no significant difference between the two preparations in the amount of barium deposited in any region of the colon (p > 0.05) or in the proportion of unsatisfactory examinations (p > 0.05). The authors conclude that the premixed suspension yielded better-quality mucosal coating than a suspension made from the dry powder and resulted in a similar high proportion of technically satisfactory studies. PMID- 8425152 TI - Translumbar aortography: experience with a steerable pigtail catheter. AB - The authors have designed a new coaxial system for translumbar aortography and studies of the pelvis and the legs from low or high aortic puncture sites. A no. 4 French pigtail catheter with a maximum flow rate of 18 mL/s is mounted coaxially on a 32-cm-long, 20-gauge, thin-walled, two-part needle. The needle cannula accepts a guide wire of diameter 0.021 in (0.53 mm), over which the catheter can be advanced craniad for aortography or caudad for arteriography of the leg. After the first injection of contrast agent, the catheter can usually be easily redirected to complete the study. Such studies were successfully completed in 73 of 79 consecutive patients; the aorta could not be cannulated in 2, and the catheter could not be redirected in 4. The sole complications were asymptomatic extravasation of the contrast agent in one patient and moderately severe back pain that resolved spontaneously in another. The authors describe the technique, as well as variations that have been developed to overcome aortic abnormalities. PMID- 8425153 TI - Cavitary aspergillosis as a complication of AIDS. AB - The authors report the radiographic findings in two patients with the human immunodeficiency virus (HIV) who presented with cavitary lung disease caused by Aspergillus. Recognition of the disease in one of the patients led to successful medical therapy. Disease due to Aspergillus must be considered in HIV-positive patients with cystic or cavitary disease appearing in chest radiographs. PMID- 8425154 TI - Primary pulmonary hypertension associated with HIV infection. AB - A Haitian woman with the human immunodeficiency virus (HIV) presented with dyspnea, cough, fatigue and lower abdominal pain of recent onset. Clinical, radiologic and hemodynamic investigations demonstrated pulmonary hypertension. The patient died a few days later. The pathological findings were compatible with primary pulmonary hypertension. This case is similar to others that have been reported and indicates a possible link between HIV infection and rapidly progressive primary pulmonary hypertension. PMID- 8425155 TI - An unusual cause of asymmetric venous engorgement detected by mammography. AB - The author reports an unusual cause of asymmetric venous engorgement, which was detected in a mammogram of an elderly patient. The anomaly was a complication of long-term hemodialysis. PMID- 8425156 TI - Delayed perforation of a duodenal diverticulum by a biliary endoprosthesis. AB - A 73-year-old woman with cholangitis due to stricture of the bile duct and the presence of stones was treated by the placement of a biliary endoprosthesis. However, 8 weeks later the lower end of the stent had retracted into a duodenal diverticulum and penetrated its inferior wall. The patient was readmitted to hospital for treatment of a retroperitoneal abscess. PMID- 8425157 TI - Optimizing the detection of colorectal liver metastases within the Canadian health care system. AB - Colorectal metastases in the liver are common. In appropriately selected patients resection can increase the 5-year survival rate from 0% to 33%. When consulted by their surgery colleagues about patient selection, radiologists now have a wide choice of imaging methods, including nuclear medicine, ultrasonography, unenhanced computed tomography (CT), intravenously enhanced CT (by infusion or by the dynamic and delayed methods), arterially enhanced CT (specifically, CT arterial portography and CT angiography) and magnetic resonance imaging. In Canada this wide range of techniques has appeared in an era of cost constraint. This article discusses the surgical background for imaging colorectal metastases in the liver and reviews the available imaging methods. The authors present an algorithmic approach that attempts to balance the need for accuracy with the logistic and financial constraints under which radiologists work in Canada. They suggest that this algorithm would allow optimal use of available resources in most Canadian hospitals. PMID- 8425158 TI - Primary malignant rhabdoid tumour of the liver. AB - The authors describe the imaging features (from radiography, ultrasonography, computed tomography and angiography) of primary malignant rhabdoid tumour of the liver in an infant. The findings were not specific, and the diagnosis of this aggressive neoplasm was based on observations obtained with electron microscopy and immunohistochemical stains. PMID- 8425159 TI - Posteromedial dislocation of the ankle without fracture or diastasis. AB - This case report describes a patient with posteromedial dislocation of the ankle without fracture and without disruption of the tibiofibular syndesmosis. The pathogenesis of this uncommon lesion is discussed. PMID- 8425160 TI - Antenatal unilateral hydrocephalus. AB - Fetal ventriculomegaly usually involves both lateral ventricles. If dilatation of the dependent lateral ventricle is seen, it might be assumed that the condition is bilateral and symmetric and that reverberation artifact is obscuring the ventricle nearer to the transducer. However, unilateral hydrocephalus can occur, though rarely and usually as a result of unilateral obstruction of the foramen of Monro. Careful attention to visualization of the obscured hemisphere is emphasized. In the patient described here the condition was associated with frontoethmoidal encephalocele. PMID- 8425161 TI - Residents' corner. Answer to case of the month #17. Synovial osteochondromatosis. PMID- 8425162 TI - Euthanasia: pro or con? PMID- 8425163 TI - RDS and surfactant replacement: the double-edged sword. PMID- 8425164 TI - Children with central lines: evidence-based practice. PMID- 8425165 TI - Nursing research and political change: the street health report. PMID- 8425166 TI - Participatory research. PMID- 8425167 TI - Applying nursing diagnosis in critical care. PMID- 8425168 TI - The sacred cow contest. PMID- 8425169 TI - Creating the impetus for research-based practice. PMID- 8425170 TI - Nurses' worklife: researching the quality. PMID- 8425171 TI - Felt learning needs of pregnant women. PMID- 8425172 TI - Using research findings in the hospital. PMID- 8425173 TI - [Biofeedback and relaxation for patients with hypertension]. AB - We have known for decades that essential hypertension significantly contributes to many physical diseases. In most cases, hypertension results from a person's perception of stress producing events or situations. According to Greenberg, when a person is confronted by a situation or an event, the person must perceive it as either positively or negatively stressful. If it is perceived as positively stressful, a series of psychological reactions are incurred followed by physiological reactions. If these psychophysiological manifestations persist, the person is then faced with possible pathological conditions. This study attempted to support the hypothesis that people with hypertension should be able to benefit from biofeedback and relaxation techniques. Using these techniques, they will be better able to recognize and understand perceived stressful events or situations and will therefore act appropriately. The purpose of this study is to examine the effects of biofeedback, relaxation and the combination of both techniques on medically diagnosed hypertensive patients. More specifically, the study examined the following questions: What is the clients' ability to lower their blood pressure using biofeedback, relaxation and the combination of both techniques? To what capacity can clients reduce their biofeedback and relaxation training programme? Twenty-three males, aged 40 to 70 years (M = 54 years), who were registered with a cardiovascular disease prevention and rehabilitation centre, volunteered to participate in this study. For five weeks, clients trained Tuesdays and Thursdays, one hour per session. Compliance to the training programme was respected by using only those clients who had missed fewer than three of the sessions. The results showed little statistical significance.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425174 TI - [In search of nursing practices]. AB - This article is based on a research begun in 1990 to gain a portrait of persons in the various holistic occupations in Quebec. From this research, the author came to question the motivation that pushed so many nurses to go into holistic practices. The goal of the research was not to collect specific data about nurses but to identify those individuals now working in holistic therapies. The author summarizes the theoretical framework of the first project and outlines the research methods used. Since the results identified those therapists with a nursing background, it was possible for her to review part of the data to identify certain characteristics of this group. The most popular holistic therapy practised was acupuncture, usually accompanied by complimentary therapies such as relaxation techniques, massage therapy, polarity and therapeutic touch. The author presents the hypothesis that nurses choose this type of practice partly to find a holistic concept that is coherent with health care, and partly to find concrete interventions compatible with traditional nursing values. She is of the opinion that it is not through this type of practice that nurses will find a solution to their professional identity problem. The study consisted of a pre tested 29 page questionnaire with a total of 89 questions sent to 954 individuals. Not all of the 89 questions were applicable to all the respondents. Of the 954 questionnaires mailed out, 11.8 percent were returned marked "incorrect address". Of the 841 therapists who received the questionnaire, 319 returned completed forms. Of these 319 respondents, 86 individuals indicated they had a nursing education. PMID- 8425175 TI - Role of tyrosine phosphorylation in radiation-induced activation of c-jun protooncogene in human lymphohematopoietic precursor cells. AB - We examined the effects of ionizing radiation on c-jun expression in human lymphohematopoietic precursors. Radiation exposure increased the level of c-jun transcripts in a dose- and time-dependent manner, providing direct evidence that ionizing radiation can activate c-jun protooncogene in human lymphohematopoietic precursors. Notable gamma-rays failed to induce c-jun expression in cells pretreated with herbimycin, and the use of cycloheximide did not overcome the inhibitory effects of herbimycin. The lack of c-jun signal in herbimycin-treated cells was not due to nonspecific damage to the distal protein kinase C signaling pathway. Thus, protein tyrosine kinase activation precedes and perhaps mandates radiation-induced activation of c-jun protooncogene expression in human lymphohematopoietic precursors. PMID- 8425176 TI - Screening for germ line p53 mutations in children with malignant tumors and a family history of cancer. AB - We have undertaken a routine investigation of the p53 status for all the children treated at our institution either affected by multiple tumors or whose family displays at least one second degree relative or less, affected by cancer before the age of 45 years. We report here on the first set of ten such families, eight of which were identified through a proband with sarcoma. p53 exons 5 to 8 have been sequenced following polymerase chain reaction amplification performed on DNA isolated from total blood. A missense mutation affecting codons 248, 273, and 282 was identified in three families. The mutation was inherited in these three families and was detected in unaffected members. In seven families no mutation was detected in exons 5 to 8. PMID- 8425177 TI - DNA-protein cross-links in welders: molecular implications. AB - A new method for detecting DNA-protein cross-links involving selective precipitation of DNA containing cross-linked proteins by K(+)-sodium dodecyl sulfate was utilized in the peripheral WBC of 21 male metal arc welders and in 26 male controls of similar age and racial characteristics who were not exposed to welding fumes. DNA was quantitated by Hoechst fluorescence. Although the concentration of nickel and chromium in the peripheral blood did not differ between subjects in the two groups, one-fourth of the welders had levels of DNA protein cross-links that were above the upper limit of the controls. Mean cross link values were 1.85 +/- 1.14% (SD) among the welders and 1.17 +/- 0.46% among the controls, a 58% statistically significant difference (P = 0.01). Thus, many welders appeared to be burdened with an excess of DNA-protein cross-links, suggesting exposure to cross-linking agents and, possibly, a detectable biological effect of potential genotoxic consequences. PMID- 8425178 TI - Interleukin 6 stimulates the production of immunoreactive endothelin 1 in human breast cancer cells. AB - To investigate the potential regulation of endothelin 1 production in human breast cancer cells, we measured the release of immunoreactive endothelin 1 (ir ET-1) from the MCF-7 and ZR-75-1 breast cancer cell lines in response to various agents including estrogen and tamoxifen as well as several cytokines. ir-ET-1 was detected in conditioned medium of MCF-7 cells and ZR-75-1 cells by specific radioimmunoassay. Among the agents tested, estrogen, tamoxifen, tumor necrosis factor, gamma-interferon, interleukin (IL) 1, and transforming growth factor beta had no effect on ir-ET-1 secretion by these breast cancer cells. However, IL-6 (20 ng/ml) treatment of MCF-7 cells and ZR-75-1 cells caused maximal increases in the amount of ir-ET-1 secreted into the culture medium to 206 and 314% of basal values after 6 h, respectively. This effect of IL-6 on ir-ET-1 secretion was inhibited by actinomycin D and cycloheximide, indicating that IL-6 stimulates de novo synthesis of ir-ET-1 at a transcriptional level. Reverse-phase high performance liquid chromatography coupled with radioimmunoassay in the conditioned medium from IL-6-treated cells revealed one major ir-ET-1 component corresponding to human standard ET-1. The present study demonstrates the potential for IL-6 to stimulate ir-ET-1 production in human breast cancer cells, which may participate in the process of acute phase reactant-like expression of this peptide and/or in the process of IL-6 enhanced breast cancer cell motility, the latter being recently clarified. PMID- 8425179 TI - p53 abnormalities in different subtypes of human sarcomas. AB - In this report we examined p53 alterations at the DNA, mRNA, and protein levels on tissue from 39 patients with different subtypes of sarcoma. Loss of heterozygosity for the chromosome 17p region was found in 60, 63, and 33% of 10 informative osteosarcomas, 11 malignant fibrous histiocytomas, and 6 leiomyosarcomas, respectively. In addition, 2 of 10 tumors belonging to a heterogeneous group of soft tissue sarcomas showed loss of heterozygosity. Elevated levels of p53 mRNA were found in six tumors, four had a truncated transcript, and in six patients no mRNA was detected. In most cases, elevated transcript levels were accompanied by overexpression of protein as studied by immunohistochemistry, whereas the presence of truncated transcripts was associated with negative immunostaining. Point mutations in exons 5, 7, or 8 of the TP53 gene were detected in seven tumors. Six of these expressed high levels of mRNA and protein, probably reflecting a point mutation in one of the alleles and loss of the other. Three of the mutations have not previously been described. Taken together, p53 abnormalities were found in approximately 65% of the osteosarcomas, malignant fibrous histiocytomas, and leiomyosarcomas examined and in 30% of the other soft tissue tumors. The results indicate that the TP53 gene is involved in the tumorigenesis of several sarcoma subtypes in a higher fraction of cases than was previously recognized. PMID- 8425180 TI - Trends in program project grant funding at the National Cancer Institute. AB - In summary, analysis of the P01 program indicates that differences exist in scores and funding requirements between purely basic and translational P01s; P01 funding has been stable and the P01 policies consistent; growth in average costs for P01s has lagged compared to R01s; P01 grantees have a higher success rate than R01 grantees; priority score compression reduces the P01 payline without reducing the number of grants funded; exception funding in P01s expands the number of grants awarded and helps to meet the scientific program needs of the NCI; almost all R01 and P01 applications in the first quartile are funded; comparably ranked R01s and P01s are funded at equivalent percentages of recommended funding levels; the percentage of approved funds awarded declines as scores for R01s and P01s become less favorable. Because the resources available are finite, the NCI carefully considers the competing demands for RPG funds. On balance, however, the P01 program is still in good health. In combination with the other methods we use to make RPG awards, the P01 program is a vital component of our research grant program. Faced with budget realities, however, some difficult decisions have to be made. The NCI is open to suggestions on how to distribute research funds to best promote cancer research throughout the country. The NCI is particularly interested in hearing researchers' perceptions of problems so that, whenever it is appropriate, we can modify our course. There are important questions to consider for the future. Should there be greater or lesser use of RFAs in allocating resources for P01 grants? What algorithm optimally distributes resources among R01s, P01s, and other funding instruments in the RPGs? Should exceptions be used more or less heavily in the P01 funding process? Should other grant programs outside the RPG budget, such as the P50 SPORE program, be used to relieve pressures on the P01 budget? Should the number of projects within P01s be limited? How can peer review groups be given opportunities to provide priority scores that better reflect distinguishable differences in highly meritorious P01s? Should P01 scores be normalized? How could the R01 percentile concept be applied to P01s? How can the results from standing study sections be synchronized with results from ad hoc review groups? What changes in peer review procedures would promote equitable score distributions and budget recommendations across years? How can the NCI provide stability for established P01s without impinging on the ability of new P01s to enter the system?(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8425181 TI - Role of the BCR-ABL oncogene in human leukemia: fifteenth Richard and Hinda Rosenthal Foundation Award Lecture. PMID- 8425182 TI - Identification of cellular defect in UVS1, a UV-sensitive Chinese hamster ovary mutant cell line. AB - UVS1 is an intermediately UV-sensitive Chinese hamster ovary mutant originally isolated by its hypersensitivity to an anticancer drug, 1-[(4-amino-2-methyl-5 pyrimidinyl)methyl]-3-(2-chloroethyl)-3-nitrosour ea hydrochloride. By cell fusion analysis, UVS1 complemented the UV sensitivity of the mouse lymphoma cell line US31 from the eighth complementation group of UV-sensitive rodent cell lines. By enzyme-linked immunosorbent assay we found that within 3 h after UV irradiation both pyrimidine dimers and (6-4)photoproducts in UVS1 were not removed from chromosomal DNA in UVS1 at all. Twenty-four h after UV irradiation the removal rate of (6-4)photoproducts was intermediate between CHO9, the parental cell line, and 43-3B, a UV-hypersensitive Chinese hamster ovary mutant of the complementation group 1, whereas the pyrimidine dimers in UVS1 were removed less efficiently as 43-3B. Alkaline elution assay showed that the incising activity to damaged DNA after UV irradiation of UVS1 was as low as that of 43-3B. The number of 1-[(4-amino-2-methyl-5-pyrimidinyl)methyl]-3-(2 chloroethyl)-3-nitrosour ea hydrochloride-induced DNA interstrand cross-links of UVS1 was almost equal to that of 43-3B and about 1.5 times more than that of CHO9, suggesting that the gene products defective in UVS1 and 43-3B are essential for the excision repair of DNA damages produced by 1-[(4-amino-2-methyl-5 pyrimidinyl)methyl]-3-(2-chloroethyl)-3-nitrosour ea hydrochloride. PMID- 8425183 TI - Clonal analysis of the expression of multiple transformation phenotypes and tumorigenicity by morphologically transformed 10T1/2 cells. AB - Seventy-five clonal populations of morphologically transformed 10T1/2 cells were established from independent Type II and Type III foci that were of spontaneous origin or were induced by the carcinogenic agents N-methyl-N'-nitro-N nitrosoguanidine, benzo(a)pyrene diol epoxide-I, and 3-methylcholanthrene. Clonal populations were characterized for expression of selected transformation phenotypes, including growth to elevated saturation density before cessation of proliferation, anchorage independence, ability to reconstruct foci when plated in the presence of wild-type 10T1/2 cells, and tumorigenicity. Forty-one % of the clonal populations expressed only the phenotype of morphological transformation, while 20% expressed all of the transformation phenotypes, including tumorigenicity, in addition to morphological transformation. The remaining clonal populations expressed varying combinations of one or more of the four transformation phenotypes. Clonal populations expressing almost all of the 16 possible combinations of the transformation phenotypes were observed, suggesting that the individual phenotypes segregated independently. Morphological transformation alone was a poor indicator of tumorigenicity, correctly predicting the tumorigenic potential of only 37% of the clonal populations. Among morphologically transformed clonal populations, coexpression of anchorage independence correctly predicted the tumorigenicity of 81% and coexpression of reconstruction of foci on a confluent lawn of wild-type cells correctly predicted the tumorigenicity of 91%. The probability that a morphologically transformed clonal population was tumorigenic correlated with the total number of transformation phenotypes expressed. Expression of the transformation phenotypes differed between tumorigenic and nontumorigenic clonal populations but not between clonal populations established from Type II and Type III foci. Tumorigenicity varied among transformed clonal populations that were induced by the different carcinogenic agents or were of spontaneous origin but did not differ between clonal populations established from Type II and Type III foci. PMID- 8425184 TI - Metabolic activation of N-hydroxy-2-aminofluorene and N-hydroxy-2 acetylaminofluorene by monomorphic N-acetyltransferase (NAT1) and polymorphic N acetyltransferase (NAT2) in colon cytosols of Syrian hamsters congenic at the NAT2 locus. AB - Acetylator genotype is regulated at the polymorphic acetyltransferase (NAT2) gene locus in humans and other mammals such as Syrian hamsters. Human slow acetylator phenotypes have been associated with increased incidences of urinary bladder cancers, whereas rapid acetylators have been associated with increased incidences of colorectal cancers. The genetic predisposition of rapid acetylators to colorectal cancers suggests localized metabolic activation of arylamine carcinogen metabolites by polymorphic N-acetyltransferase (NAT2) in colon tissues. We tested this hypothesis in Bio. 82.73/H Syrian hamster lines which are congenic at the NAT2 gene locus. Congenic Bio. 82.73/H Syrian hamsters expressed acetylator genotype-dependent N-acetyltransferase activity in colon cytosols toward arylamine carcinogens such as 2-aminofluorene and 4-aminobiphenyl. Partial purification of the hamster colon cytosol by anion exchange chromatography identified two N-acetyltransferase isozymes analogous to those previously described in liver and urinary bladder. One of the isozymes (NAT2) exhibited acetylator genotype-dependent expression for the N-acetylation of each arylamine tested: p-aminophenol; 2-aminofluorene; 4-aminobiphenyl; 3,2'-dimethyl-4 aminobiphenyl; and 2-amino-dipyrido[1,2-a:3',2'd]imidazole as well as for the metabolic activation (via O-acetylation) of N-hydroxy-2-aminofluorene to form DNA adducts. Although NAT2 catalyzed the metabolic activation of N-hydroxy-2-acetyl aminofluorene to DNA adducts, the rates were lower, were paraoxon-sensitive, and did not reflect acetylator genotype. A second isozyme (NAT1) also catalyzed the N acetylation of each arylamine as well as the metabolic activation of N-hydroxy-2 aminofluorene and N-hydroxy-2-acetylaminofluorene to DNA adducts at rates that were independent of acetylator genotype. Metabolic activation of N-hydroxy-2 aminofluorene catalyzed by both NAT1 and NAT2 was resistant to 100 microM paraoxon, an inhibitor of microsomal deacetylases. Metabolic activation of N hydroxy-2-acetylaminofluorene by NAT1 and NAT2 was partially sensitive to 100 microM paraoxon. Michaelis-Menten kinetic constants were determined for the colon NAT1 and NAT2 isozymes and compared to previous determinations for liver NAT1 and NAT2. For each of the arylamines tested, both apparent Km and apparent Vmax were higher for NAT2 than NAT1. In rapid acetylator hamster colon, NAT2/NAT1 activity ratios were 18 and 13 for the N-acetylation of 2-aminofluorene and 4 aminobiphenyl and 28 for the O-acetylation of N-hydroxy-2-aminofluorene. These results strongly support the role of the polymorphic NAT2 gene locus in the local metabolic activation of N-hydroxyarylamine carcinogens in colon and provide mechanistic support for human epidemiological studies suggesting a predisposition of rapid acetylators to colorectal cancer. PMID- 8425185 TI - Vegetables, fruit, and lung cancer in the Iowa Women's Health Study. AB - Previous epidemiological studies have shown an inverse association between vegetable and fruit consumption and lung cancer risk; few of these studies have been prospective or have focused upon women. In 1986, we assessed food intake in 41,837 Iowa women, aged 55 to 69 yr, with a mailed 127-item food frequency questionnaire. After 4 years of follow-up, 179 incident cases of lung cancer were identified via the Iowa Surveillance, Epidemiology, and End Results cancer registry. After specific exclusion criteria were applied, a nested case-control comparison of 138 cases with 2,814 randomly selected noncases was undertaken. Intakes, in the upper-most quartile, of 11 vegetable and fruit groups, as well as of the nutrients beta-carotene and vitamin C, were explored. High intakes of all vegetables and fruit, all vegetables, and green leafy vegetables were each associated with an approximate halving of risk: age-, smoking-, and energy adjusted odds ratios (ORs) were 0.49 (95% confidence interval, 0.28-0.86), 0.50 (95% confidence interval, 0.29-0.87), and 0.45 (95% confidence interval, 0.26 0.76), respectively. A lower lung cancer risk was also seen for all fruit (adjusted OR = 0.75 for high consumption), high vitamin C vegetables and fruit (OR = 0.75), carrots (OR = 0.71), and brocolli (OR = 0.72) and for the nutrients beta-carotene (OR = 0.81) and vitamin C (OR = 0.81) (all 95% confidence intervals included 1.0). Lung cancer risk was unrelated to consumption of the three food groups defined as "high-carotenoid" (beta-carotene, lutein, and lycopene) and tomatoes. In an analysis stratified by histological type of lung cancer, the strongest inverse associations for vegetables and fruit were seen for large cell carcinoma. Analysis by smoking status showed the inverse associations for most vegetable and fruit groups with lung cancer risk to be stronger for exsmokers than for current smokers. Results from the stratified analyses must be interpreted with caution because of the small number of cases in each stratum. PMID- 8425186 TI - Reduced glutathione protects against cisplatin-induced neurotoxicity in rats. AB - Reduced glutathione (GSH) is reported to diminish cisplatin-induced neurotoxicity, and it was for this reason that we studied GSH in an animal model of cisplatin neuropathy. The neuropathy was evaluated by measuring the sensory nerve conduction velocity (SNCV) in young adult Wistar rats. GSH injections (i.v.) were given twice weekly, within five minutes before cisplatin was injected (i.p.). In a first experiment GSH (500 mg/kg) in combination with a low-dose cisplatin treatment (1 mg/kg, 10 weeks) was investigated. Animals treated with cisplatin and placebo developed a neuropathy (SNCV at week 10: age controls, 61.9 m/s; cisplatin alone, 44.2 m/s), whereas rats treated with cisplatin and GSH did not (SNCV, 61.9 m/s). The same dose of GSH was used in combination with a high dose cisplatin schedule (2 mg/kg, 5 weeks' treatment plus 5 weeks' recovery). Again, GSH protected animals against the development of neuropathy (SNCV at week 10: age controls, 61.9 m/s; cisplatin alone, 50.6 m/s; cisplatin plus GSH, 61.1 m/s). In another experiment four lower doses of GSH (25, 50, 100, and 200 mg/kg) were tested in combination with the low-dose cisplatin protocol (1 mg/kg, 11 weeks). The cisplatin group developed a neuropathy (SNCV at week 11: cisplatin alone, 50.2 m/s; age controls, 60.6 m/s). Only the dose of 200 mg GSH/kg was found to protect against the development of a neuropathy (SNCV, 61.0 m/s). In an antitumor study GSH administered at 300 mg/kg in combination with cisplatin at 1.5 mg/kg did not diminish the curative effect of cisplatin. We conclude that GSH prevents cisplatin-induced neuropathy and that it should be investigated further in the clinic. PMID- 8425187 TI - Characterization of experimental mitoxantrone cardiotoxicity and its partial inhibition by ICRF-187 in cultured neonatal rat heart cells. AB - Mitoxantrone cardiotoxicity was investigated in isolated neonatal rat heart myocytes treated for 3 h in the presence or absence of the metal chelator ICRF 187. Electron microscopy studies of mitoxantrone-treated myocytes showed disorganized myofibrillar structures, swollen mitochondria, and extensive vacuolization. Cardiotoxicity, reflected as the ratio of intracellular ATP/protein and the loss of spontaneous beating, was only partially reduced by continuous ICRF-187 concentrations up to 50 micrograms/ml. ICRF-187 induced myocyte protection which was dependent on the dose and duration of exposure. ICRF 187 also reduced the cardiotoxicity of doxorubicin to a lesser extent and reduced the toxicity of a postulated cyclic mitoxantrone metabolite. However, the cardiotoxicity of ametantrone, a nonmetal-binding analogue of mitoxantrone, was unaltered with ICRF-187. The antitumor activity of mitoxantrone was unaltered by ICRF-187 in human tumor cells and in P-388-bearing mice. In addition, ICRF-187 allowed for 50% greater cumulative dosing in normal mice that, nonetheless, showed extensive histological heart damage 7 wk after dosing. These studies show that ICRF-187 provides partial protection from mitoxantrone cardiotoxicity in vitro without impairing the drug's antitumor activity in vitro or in vivo. This facilitates greater cumulative drug dosing in normal mice. The postulated mechanism of cardioprotection is metal chelation, because ICRF-187 did not alter the toxicity of a nonchelating mitoxantrone analogue. PMID- 8425188 TI - Nitric oxide/nucleophile complexes inhibit the in vitro proliferation of A375 melanoma cells via nitric oxide release. AB - Cell-mediated antitumor effects have, in part, been attributed to the production of NO. Compounds which generate NO might, therefore, be useful in attenuating the growth of tumor cells. Six nitric oxide/nucleophile adducts that release NO spontaneously in solution were tested for their effectiveness in inhibiting DNA synthesis in A375 human melanoma cells. The complexes of NO with spermine, 3-(n propylamino)propylamine (PAPA/NO), and diethylamine reduced thymidine incorporation by 50% at concentrations of 24, 44, and 128 microM, respectively. The degree of inhibition was, in general, related to the rate and extent of NO release in solution. A melanoma cell clone sensitive to interleukin 1-mediated cytostasis (A375-C6) was no more sensitive to PAPA/NO than a clone resistant to interleukin 1 (A375-C5), suggesting that the differing inhibitory effects of interleukin 1 in the two A375 cell clones are not due to a differential sensitivity to nitric oxide. Oxymyoglobin (125 microM), a known scavenger of NO, restored the ability of A375-C6 cells to incorporate thymidine in the presence of up to 200 microM PAPA/NO. When PAPA/NO was added to a solution of oxymyoglobin, nitrosylmyoglobin was formed, indicating that the protective effect of myoglobin was due to scavenging of NO. The results are consistent with a nitric oxide mediated mechanism for NO/nucleophile cytostasis and suggest that such compounds may be useful as tools for investigating the role of reactive nitrogen intermediates in cytostasis and cytotoxicity. PMID- 8425189 TI - Combined interleukin 1 beta/interleukin 2 treatment in mice: synergistic myelostimulatory activity and protection against cyclophosphamide-induced myelosuppression. AB - We have studied the effects of single and combined treatment with interleukin 1 beta (IL-1 beta) and interleukin 2 (IL-2) on spleen and bone marrow hematopoiesis in normal mice. Injection of IL-1 beta alone was followed by a significant increase in the number of granulocytes in spleen and progenitors (burst-forming units-erythroid and colony-forming units-granulomonocytic) in both spleen and bone marrow, s compared to control mice. In contrast, IL-2 alone induced only a slight increase in the number of marrow colony-forming units-granulomonocytic and had no significant effect on spleen progenitors. Repeated injections of both IL-1 beta and IL-2 resulted in a synergistic increase in spleen weight and splenocyte number, as compared to mice treated with the single cytokine regimen; in particular, the combined treatment induced a marked rise in the number of neutrophilic granulocytes and erythroblasts, whereas splenic lymphocytes were not affected. This regimen also caused a synergistic increase in the number of spleen and marrow progenitor cells: a time-course analysis showed an elevation in numbers of both burst-forming units-erythroid and colony-forming units granulomonocytic, first in marrow (day 10) and subsequently in spleen (day 18). Combined IL-1 beta/IL-2 treatment dampened the decrease and accelerated the recovery of myeloid cells after cyclophosphamide injection, whereas the single cytokine regimen was not effective. Similarly, the rebound of WBC (especially neutrophilic granulocytes) after cyclophosphamide treatment was markedly enhanced by the combined treatment, whereas the single cytokine regimen was ineffective. These results, indicating a myelostimulatory effect by the combined cytokine regimen, together with our previous observations showing a synergistic antitumor activity by IL-1/IL-2 treatment in experimental mouse tumors (V. Ciolli et al., J. Exp. Med., 173: 313-322, 1991), may provide the basis for the development of new combination therapies with cytokines and antiblastic agents in the treatment of cancer patients. PMID- 8425190 TI - Antisense DNA inhibition of tumor growth induced by c-Ha-ras oncogene in nude mice. AB - Antisense DNA has shown an ability to target specific oncogene transcripts and inhibit their expression in cells, but the degree to which sustained treatment can suppress total levels of an oncogenic product and alter tumorigenesis in vivo remains to be determined. In this study, NIH-3T3 cells transformed by the activated c-Ha-ras oncogene from T24 human bladder cancer cells were treated for 3 consecutive days in vitro with an antisense DNA pentadecamer complementary to a target in the 5'-flanking region of the c-Ha-ras RNA transcript. Following antisense DNA treatment, a portion of the cells was lysed for measurement of RAS p21 while the remaining cells were evaluated for tumorigeneity by injection s.c. into athymic nude mice at a dose of 5 x 10(5) cells/mouse. The 3 days of treatment with the anti-c-Ha-ras DNA reduced RAS p21 cellular levels by more than 90% while a nonspecific control DNA reduced p21 levels by approximately 20%. Tumor growth of cells treated with anti-c-Ha-ras DNA was significantly reduced for up to 14 days following the end of treatment and implantation into the mice whereas the nonspecific control DNA had no significant effect. These effects on tumor growth were evident in two different strains of nude mice and in both males and females. It is suggested that the pronounced decrease in RAS p21 levels produced by anti-c-Ha-ras DNA resulted in a reversal of the transformed phenotype, and it is this reversal which accounts for the prolonged inhibition of tumorigenesis following antisense DNA treatment. PMID- 8425191 TI - Antitumor activity of N1,N11-bis(ethyl)norspermine against human melanoma xenografts and possible biochemical correlates of drug action. AB - In in vitro systems, the spermine analogue, N1,N11-bis(ethyl)norspermine (BENSPM), suppresses the polyamine biosynthetic enzymes, ornithine and S adenosylmethionine decarboxylase (ornithine decarboxylase and S adenosylmethionine decarboxylase, respectively), greatly induces the polyamine catabolic enzyme, spermidine/spermine N1-acetyltransferase (SSAT), depletes polyamine pools, and inhibits cell growth. Against MALME-3 M human melanoma xenografts, BENSPM and related homologues demonstrate potent antitumor activity that has been found to correlate positively with their ability to induce SSAT activity in vitro. Herein, we further evaluate the antitumor activity of BENSPM and at the same time characterize the biochemical effects of BENSPM treatment on polyamine metabolism of selected normal and tumor tissues. At 40 mg/kg 3 times/day for 6 days i.p., BENSPM suppressed growth of MALME-3 M human melanoma xenografts during treatment and for 65 days afterwards. Similar antitumor activity was obtained with 120 mg/kg once daily for 6 days and 40 mg/kg once daily for 6 days, indicating that against this tumor model, the dosing schedule can be relaxed up to sixfold without compromising antitumor activity. When MALME 3 M tumor-bearing mice were retreated with BENSPM 2 weeks after the first treatment at 40 mg/kg 3 times/day for 6 days, initial tumor volumes of 85 mm3 were reduced to < 10 mm3. Analysis of melanoma, liver, and kidney tissues from mice treated with 40 mg/kg 3 times/day for 6 days revealed relatively similar accumulations of BENSPM in all tissues at levels greater than the original total content of polyamine pools. By 2 weeks following treatment, BENSPM pools in normal tissues were almost gone, whereas in tumor tissues significant amounts (40%) were still retained. The biosynthetic enzymes, ornithine decarboxylase and S-adenosylmethionine decarboxylase, gave no indication of enzyme suppression (or increase) by the analogue as typically occurs in vitro. By contrast, SSAT was induced from an average of < 50 pmol/min/mg in control tissues to 320 pmol/min/mg in liver, 1255 pmol/min/mg in kidney, and 13,710 pmol/min/mg in MALME-3M tumor. Two weeks later, SSAT activity was still 12 times higher in tumor than in kidney. Polyamine pools (putrescine, spermidine, and spermine) were reduced after treatment in all tissues and approached near-total depletion in the tumor. Good antitumor activity and even more potent induction of SSAT (i.e., 26,680 pmol/min/mg) was also observed in PANUT-3 human melanoma xenografts. Overall, the findings reveal meaningful antitumor activity by BENSPM against 2 human melanoma xenografts and provide in vivo evidence consistent with SSAT-induced polyamine depletion playing a determining role in at least the initial phase of the antitumor response. PMID- 8425192 TI - Markedly improved efficacy of edatrexate compared to methotrexate in a high-dose regimen with leucovorin rescue against metastatic murine solid tumors. AB - 10-Ethyl-10-deazaaminopterin (EDX, edatrexate) exhibits therapeutic activity against methotrexate (MTX)-resistant tumors in animals and patients. In an effort to improve its efficacy among more chemoresistant tumors, studies were initiated in murine models of advanced metastatic disease comparing EDX and MTX at their maximum tolerated dose alone and in a high-dose regimen incorporating low-dose, delayed Ca leucovorin (LCV) rescue. Both twice-weekly x 3 and weekly x 3 schedules of administration were used with LCV given 16, 20, and 24 h after EDX. The LCV dose required to protect mice was 1/40 and 1/20 of the EDX or MTX dose, respectively, on either schedule. Therapy was initiated 5 or 6 days following i.v. implant of 5 x 10(5) cells of the E0771 mammary adenocarcinoma, T241 fibrosarcoma, Lewis lung carcinoma, B16 melanoma, or C38 colon carcinoma. MTX was essentially ineffective (increase in life span = < 30%) when given alone and either ineffective or only modestly effective (increase in life span = 20-80%) in increasing survival when given in the high-dose regimen to tumor-bearing mice. EDX alone was more effective than MTX when it was given in either regimen of therapy. Also, EDX given in the high-dose regimen (either twice-weekly or weekly x 3) was markedly more effective than EDX alone. Increased survival with this regimen was 2-3-fold greater than EDX alone against all 5 tumors, and long-term survivors were obtained with E0771 (20%), T241 (30-40%), Lewis lung (10-15%), B16 (20%), and C38 (40%) tumors. The administration of 6 doses rather than 3 doses on the twice-weekly schedule against T241 and Lewis lung tumors required a modest increase in the LCV dose but substantially improved efficacy, with as much as 70% long-term survivors (T241 tumor). We conclude that the use of a high-dose regimen with delayed LCV rescue markedly improved the therapeutic effectiveness of EDX against advanced metastatic disease in tumor-bearing mice. These studies should provide a framework for further clinical work with EDX, using this modality of therapy. PMID- 8425193 TI - Molecular basis of complement resistance of human melanoma cells expressing the C3-cleaving membrane protease p65. AB - The molecular mechanism of complement resistance of the human SK-MEL-170 melanoma cell line was investigated. The cells have been shown to express the C3b-cleaving membrane protease p65. To delineate the molecular consequences of the C3b cleaving activity for the complement cytotoxicity, the molecular events during the initiation (R24 monoclonal antibody, C1), amplification (C4, C3), and membrane attack (C5, C9) phases of complement were studied in comparison to a complement-susceptible human melanoma line (SK-MEL-93-2). No cleavage of C4b and C5b, 2 molecules structurally similar to C3b, was observed on the cells during classical pathway activation indicating the specificity of the p65 protease for the C3b molecule. The rapid degradation of C3b by p65 on the surface of complement-resistant SK-MEL-170 cells generates a M(r) 30,000 C3 alpha'-chain fragment detectable as early as 1 min after complement activation, whereas no such fragment was present in detectable amounts on complement-susceptible cells. As a result of the rapid C3b proteolysis by p65 on resistant SK-MEL-170 cells, less C5 convertases are formed, which in turn results in the formation of a lower number of terminal complement components and membrane attack complexes. R24 antibody and C1q binding to the resistant cells was slightly lower as to susceptible cells. C4 binding studies, however, revealed that the observed difference in antibody and C1q binding has no influence on the complement resistance of SK-MEL-170 cells: significantly more C4b was bound to complement resistant (1565 +/- 92 fg/cell) as compared to susceptible cells (715 +/- 31 fg/cell). On extraction of the molecular forms of C4 bound to the cell membranes, an additional high molecular weight C4 species--apparently a C4b-C4b homodimer- appeared only on the resistant SK-MEL-170 cells that may function as a residual back-up C5 convertase. Collectively, these results show that SK-MEL-170 human melanoma cells evade complement-mediated cytolysis despite sufficient activation of early components of the classical complement pathway by p65-mediated rapid degradation of surface-bound C3b, leading to a significant reduction in membrane attack complex formation. Thus, rapid cleavage of surface deposited C3b was established as a powerful mechanism of complement resistance. PMID- 8425194 TI - Improved radioimmunotherapeutic efficacy of an anticarcinoma monoclonal antibody (131I-CC49) when given in combination with gamma-interferon. AB - The moderately differentiated human colon tumor cell line, HT-29, constitutively expresses low levels of the high molecular weight mucin, tumor-associated glycoprotein 72 (TAG-72), and the M(r) 180,000 carcinoembryonic antigen (CEA) when grown as s.c. tumors in athymic mice. We report that the in vivo administration of gamma-interferon (IFN-gamma) resulted in a time- and dose dependent increase in both TAG-72 and CEA expression in the HT-29 tumors. Immunohistochemical staining revealed a more homogeneous TAG-72-positive tumor cell population after IFN-gamma. Furthermore, both anti-TAG-72 and anti-CEA monoclonal antibodies (MAbs) showed enhanced localization to the HT-29 tumors in mice treated with IFN-gamma. Using that experimental model, subsequent studies presented evidence showing that the combination of IFN-gamma with 131I-CC49, an anti-TAG-72 MAb, resulted in a statistically significant improvement in therapeutic efficacy when compared with 131I-CC49 alone. For example, treatment with 300 microCi of 131I-CC49 initially suppressed HT-29 tumor growth; however, that reduction in tumor growth was transient as evidenced by the emergence of additional tumor growth at later time points. In contrast, an 8-day treatment with IFN-gamma in combination with 300 microCi 131I-CC49 resulted in sustained suppression of HT-29 tumor growth. Thus, IFN-gamma in vivo can substantially increase the TAG-72 expression in human colon tumor xenografts which leads to an increased tumor localization of anti-TAG-72 MAbs and seems to be responsible for the enhanced antitumor effects when IFN-gamma was combined with 131I-CC49. The results provide further evidence for including a biological response modifier, such as IFN-gamma, which can increase the expression of specific tumor antigens (i.e., TAG-72 and CEA) subsequently leading to a dramatic improvement in the antitumor efficacy of a radionuclide-conjugated MAb. PMID- 8425195 TI - Hypersensitivity of human lymphocytes to UV-B and solar irradiation. AB - T-lymphocytes from three normal human donors, irradiated with broad-spectrum UV-B (peak emission, 312 nm), are 20-fold more sensitive than fibroblasts from four normal donors in a clonogenic assay. We have compared the formation of thymine cyclobutane dimers and pyrimidine-(6-4)-pyrimidone photoproducts following irradiation by UV-C (254 nm) and UV-B and studied killing at doses giving equal dimer formation. UV-B killing of fibroblasts appears to be associated with dipyrimidine photoproduct formation, whereas UV-B killing of lymphocytes is mediated by nondimer damage. Strand breakage following UV-B irradiation measured using the "Comet" assay (single cell gel electrophoresis) reflects this nondimer damage and has kinetics consistent with excisable damage. Lymphocytes from three excision-deficient xeroderma pigmentosum donors show reduced strand breakage and increased killing following UV-B irradiation, compared with lymphocytes from normal donors. We therefore suggest that UV-B kills human lymphocytes by excisable nondimer damage and that xeroderma pigmentosum lymphocytes are defective in its repair. The putative nondimer damage does not appear to be associated with radical attack, and the strand breakage is not a manifestation of apoptosis. A 1-min exposure of human lymphocytes in vitro to natural sunlight is sufficient to produce damage measurable by the Comet assay. PMID- 8425196 TI - Tumor necrosis factor alpha and interleukin 1 alpha induce anchorage independence in v-jun transgenic murine cells. AB - The oncogene jun encodes a transcription factor of the AP-1 family. In mice carrying viral jun (v-jun) as a transgene, wounding is a prerequisite for tumorigenesis, suggesting collaboration between the transgene and a wound-related event. To define possible candidates for this collaborative process, we examined the effect of several wound-related polypeptide growth factors on cells from transgenic mice. Tumor necrosis factor alpha and interleukin 1 alpha induce anchorage independence in embryo fibroblasts and tumor cell revertants from these mice. This effect was specific for the two cytokines and was restricted to cells from v-jun transgenic mice. Anchorage independence required the continued presence of the cytokines. Transfection of transgenic cells with a v-jun expression plasmid also induced anchorage independence and a tumorigenic phenotype in transgenic tumor cell revertants. However, there was no correlation between anchorage independence, expression of Jun, and AP-1 activity. These results suggest that while increased transgene expression can enhance the growth properties of v-jun transgenic cells, there exist other cytokine-dependent mechanisms that have a similar effect. Retinoic acid, dexamethasone, or forskolin inhibits induction of anchorage independence by tumor necrosis factor alpha, interleukin 1 alpha, and transfected v-jun. Although these agents affect both AP 1 transactivation potential and DNA binding in the transgenic cells, the changes are not correlated with the inhibition of growth. PMID- 8425197 TI - Characterization of sarcoma cell lines from v-jun transgenic mice. AB - Wounding is a prerequisite for tumor formation in v-jun transgenic mice. The progression from wound to dermal sarcoma is a multistep process which, at some stage, results in an increase in transgene mRNA expression in tumor tissue. However, transgene expression in individual sarcoma cells stained for Jun protein cultures is heterogeneous. We cloned several cell lines from wound-related v-jun transgenic tumors to determine whether a relationship existed between the cellular growth properties and structure, expression, or function of the transgene. Cell lines with very high v-jun expression had a high cloning efficiency in soft agar and tumorigenicity in nude mice. However, for cell lines with an intermediate or low level of transgene expression there was no correlation between transgene expression and the transformed phenotype. There was also no correlation between transgene expression and individual cell line morphologies, growth rates, transgene genomic DNA copy number, or mRNA expression of jun-related genes. The tumor cell subclones (1-20.2, 3-24.3) with very low transgene expression, very poor cloning efficiency, and low tumorigenicity also showed reduced activator protein 1 DNA binding activity and had an increased expression of endogenous c-jun when compared to other tumor cell lines. Transfection of a v-jun expression vector into cell lines with poor cloning efficiency and low tumorigenicity enhanced both in vitro cloning and in vivo tumor formation. However, such overexpressed v-jun had no effect on NIH3T3 cells. Our studies show that expression of the v-jun transgene contributes to the transformed phenotype of tumor cell lines but that there are additional factors that determine growth properties in culture and in the animal. PMID- 8425198 TI - Differential isolation of normal luminal mammary epithelial cells and breast cancer cells from primary and metastatic sites using selective media. AB - The present studies were aimed at determining if the use of a cell culture medium that supports proliferation of human mammary epithelial cells of the luminal lineage would allow routine isolation of breast cancer cells from primary and metastatic tumor specimens. Results obtained with mammary epithelial cells derived from reduction mammoplasty specimens and primary breast carcinomas indicated that growth of cells on type I collagen-coated dishes in Ham's F-12 medium supplemented with insulin, hydrocortisone, epidermal growth factor, cholera toxin, and 5% fetal bovine serum resulted in the growth and serial passage of cells that stained positively for the luminal cell marker cytokeratin 19. By contrast, growth of mammary epithelial cells in a growth factor supplemented serum-free medium resulted in the emergence of mammary epithelial cell colonies that were uniformly negative for keratin 19. Filter isolation methods were used to isolate individual keratin-19-positive colonies from primary cultures derived from breast cancer specimens. All of the luminal mammary epithelial cells isolated from breast cancer tissues expressed characteristics of normal cells. Keratin-19-positive colonies isolated from several different tumors all grew rapidly for 30 to 60 days in culture and then senesced. Cells were isolated from one tumor that was known to have undergone a loss of heterozygosity at a specific locus in the p53 gene. All colonies isolated from this specimen contained both p53 alleles, which was consistent with their origin from normal luminal cells. Cells were also isolated from one tumor in which the c-erbB2 protein was drastically overexpressed in the neoplastic cells. Once again, keratin-19-positive colonies isolated from this tumor did not overexpress the c erbB-2 protein. Experiments were then performed with cells derived from pleural effusions and metastatic lymph nodes. Results obtained with these specimens indicated that the growth conditions that support the growth of normal luminal mammary epithelial cells do not support the growth of neoplastic cells. However, the omission of cholera toxin, epidermal growth factor, and type I collagen substratum resulted in the isolation of two long-term cell lines. Both cell lines have population doubling times of approximately 100 h, are hyperdiploid, and stain positively for cytokeratin 19. Thus, culture conditions that support the growth of normal luminal mammary epithelial cells do not, in general, support the growth of breast cancer cells. PMID- 8425199 TI - Growth inhibition of a human colorectal carcinoma cell line by interleukin 1 is associated with enhanced expression of gamma-interferon receptors. AB - Recombinant human tumor necrosis factor and recombinant human gamma interferon (IFN-gamma) exert synergistic growth inhibitory effects in WiDR human colorectal carcinoma cells. In this cell line, tumor necrosis factor increases IFN-gamma binding. Interleukin 1 (IL-1) is a cytokine that mimics many of the biological actions of TNF. Therefore, in the present study, we investigated the effects of recombinant human IL-1 on cell growth and IFN-gamma receptor expression in WiDR cells. IL-1 slightly inhibited the growth of WiDR cells, and exerted additive growth inhibitory effects in the presence of IFN-gamma. IL-1 caused a time- and dose-dependent increase in 125I-labeled IFN-gamma binding that was maximal at 6 h, persisted for at least 24 h, and was blocked by both actinomycin D and cycloheximide. The increase in binding was associated with an increase in cell surface IFN-gamma receptor protein expression as determined by Scatchard analysis of equilibrium binding data and by immunofluorescent staining with an anti-human IFN-gamma receptor monoclonal antibody. IL-1 also produced a time- and dose dependent increase in IFN-gamma receptor mRNA levels that was maximal at 3 h and persisted for at least 24 h. Actinomycin D, but not cycloheximide, completely blocked the IL-1-mediated increase in IFN-gamma receptor mRNA levels. However, IL 1 did not alter IFN-gamma receptor mRNA half-life. These data indicate that IL-1 and IFN-gamma exert additive growth inhibitory effects on colon cancer cell growth, and suggest that IL-1 increases IFN-gamma receptor expression in these cells by enhancing IFN-gamma mRNA levels. PMID- 8425200 TI - Distribution of colon cancer cells permanently labeled by lectin-mediated endocytosis of a trap label. AB - A method was elaborated for high-yield 125I-trap labeling of rat colon carcinoma cells using conjugates of dichlorotriazine aminofluorescein and bovine serum albumin substituted with either N-acetylgalactosamine or N-acetylglucosamine as vehicles. Fluorescence microscopy revealed that the ligands accumulated in perinuclear vesicles that were probably lysosomes. Monensin inhibited accumulation by 40%, signifying receptor-mediated endocytosis. Competition experiments revealed that the same receptor(s) mediated endocytosis of the two neoglycoproteins. Accumulation of label was greatly enhanced in the absence of serum, resulting in a labeling efficiency of at least 15 cpm/cell, with no sign of toxic effects. At least 75% of the initially accumulated radioactivity resided in the cells 4 days after labeling. After that the loss of radioactivity was linear with time and stabilized at 1.1%/day for at least 2 weeks. Injection of labeled carcinoma cells i.v. into syngeneic rats revealed a very rapid clearance from the circulation. Isolation of the liver cells 24 h later revealed that a great proportion of the administered cells or their remnants had been engulfed by sinusoidal Kupffer and endothelial cells; the parenchymal cells contained a smaller proportion of label. In conclusion, we have developed a technique of labeling colon carcinoma cells with 125I and fluorescein utilizing specific lectin-like receptors for endocytosis. Since the label is trapped intralysosomally, it will also label Kupffer cells and other members of the reticuloendothelial system after internalization. These features make the procedure well suited for studies on the fate of the colon carcinoma cells after administration in vivo. Since the label is trapped intralysosomally for an extended length of time, parameters such as the formation of metastasis and elimination by phagocytosis can readily be determined. PMID- 8425201 TI - Growth factor dependency and gene expression in preneoplastic mouse mammary epithelial cells. AB - The TM preneoplastic mammary outgrowth lines were established in vivo from mammary epithelial cell lines and have been characterized with respect to their tumorigenic, morphological, and functional properties. The TM outgrowth lines were then established as in vitro cell lines. A comparison of the growth factor dependencies of the TM preneoplastic lines and their progenitor cell lines grown in monolayer cell culture indicated that the TM preneoplastic cell lines exhibited a decreased dependence on epidermal growth factor for growth in vitro. The exception to this result was the TM3 cell line which still exhibited a marked dependence on epidermal growth factor for growth. An examination of several genes for mRNA levels indicated that the expression of c-neu, c-H-ras, c-myc, and retinoblastoma was not elevated in those TM preneoplasias which exhibited a decreased dependence on epidermal growth factor. Additionally, there was no evidence that c-H-ras was activated in the preneoplastic outgrowths or tumors. In contrast, mouse mammary tumor virus long terminal repeat mRNA was increased in preneoplasias and tumors, whereas gelsolin mRNA was decreased in tumors but not in preneoplasias. The down-regulation of gelsolin mRNA is consistent with recent reports in human breast cancers. These results together with those reported in another paper (D. Medina et al., Cancer Res., 53: 663-667, 1993) indicate that the TM3 outgrowth line is a minimally deviated preneoplasia which does not share many of the molecular alterations exhibited in tumorigenic TM preneoplastic outgrowth lines. These data, along with other recent data, are interpreted in a hypothesis which views the three essential characteristics of the mammary preneoplastic state as independent and dissociable genetic alterations. Importantly, each of the three characteristics is independently isolated in one or more of the in vivo outgrowth populations. These outgrowth lines should allow identification of the nature and function of the molecular alterations associated with each stage of mammary preneoplasia. PMID- 8425202 TI - Functional expression of interleukin 2 receptor in a human factor-dependent megakaryoblastic leukemia cell line: evidence that granulocyte-macrophage colony stimulating factor inhibits interleukin 2 binding to its receptor. AB - Human interleukin 2 (IL-2) is a member of the class of crucial regulators of lymphocyte proliferation. The action of IL-2 is known to be mediated through binding to a specific IL-2 receptor (IL-2R) which comprises at least two distinct proteins: IL-2R alpha (p55) and IL-2R beta (p70-75). However, the expression and function of IL-2R are largely unknown in acute myeloblastic leukemia cells. In a human granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-3, or stem cell factor-dependent myeloid leukemia cell line (M07E), IL-2 was found to stimulate proliferation in a dose-dependent manner and to augment GM-CSF- and stem cell factor-induced proliferation of M07E cells. The expression of IL-2R beta on M07E cells was detectable with 125I-IL-2 binding and affinity cross linking analyses and with a monoclonal antibody against IL-2R beta, Mik-beta 1. Although the expression of IL-2R beta was not down-regulated but somewhat up regulated by treatment with GM-CSF in both mRNA and protein levels, GM-CSF was found to compete (75%) with radiolabeled IL-2 for binding to IL-2R on M07E cells, whereas no competition of GM-CSF binding was observed with IL-2 even at a 400 fold molar excess. These results suggest that IL-2R may be functionally expressed in some cases of acute myeloblastic leukemia cells and raise the possibility that IL-2 may have some effects on human myelopoiesis. PMID- 8425203 TI - Chromosomes 8, 12, and 17 copy number in Astler-Coller stage C colon cancer in relation to proliferative activity and DNA ploidy. AB - Fluorescence in situ hybridization using centromere-specific DNA probes to chromosomes 8, 12, and 17 was applied to 23 archival paraffin-embedded stage C colonic cancer specimens. Chromosome copy number was related to flow cytometric determinations of S-phase fraction and DNA ploidy. Three to eight copies of chromosomes 8, 12, and 17 were observed at mean frequencies of 28.7%, 37.8%, and 20.9%, respectively. The mean frequency of multiple copies of chromosome 12 was significantly greater than that for chromosome 17 (P < 0.0025). The mean frequency of single copies of chromosome 17 was significantly greater than that for chromosomes 8 and 12 (P < 0.0025 and P < 0.0005, respectively). Regarding the fourth quartile of cases, defined on the basis of the frequency of multiple chromosome copies, the proportion demonstrating moderate to high proliferative activity greatly exceeded the proportion displaying low proliferative activity. The same cases (most chromosomally aberrant) also generally demonstrated DNA aneuploidy. The results indicate a substantial degree of karyotypic instability in advanced colon cancer, particularly in cases with high proliferative activity and DNA aneuploidy. PMID- 8425204 TI - Milky spots as the implantation site for malignant cells in peritoneal dissemination in mice. AB - We examined the site-specific implantation of cancer cells in peritoneal tissues after an i.p. inoculation of 10(5) P388 leukemia cells. Twenty-four h after the inoculation, the number of viable cancer cells infiltrating into specific tissue sites of the peritoneum was estimated by an i.p. transfer method. A descending order of tissue implantation with cancer cells was established as omentum > gonadal fat > mesenterium > posterior abdominal wall > stomach, liver, intestine, anterior abdominal wall, and lung. A significant correlation was established between the logarithm of the number of infiltrating cancer cells and the logarithm of the number of milky spots. Next, the omentum was examined microscopically after i.p. inoculation with P388 leukemia cells labeled with bromodeoxyuridine or B-16 PC melanoma, which were differentiated from the other cells by an immunocytological method using anti-bromodeoxyuridine antibody or by the melanin of the B-16 PC melanoma cells. These cancer cells were found microscopically to be infiltrating only the milky spots, whereas none were seen at the other sites. These results suggest that cancer cells seeded i.p. specifically infiltrate the milky spots in the early stage of peritoneal metastases. PMID- 8425205 TI - Melanoma-mediated dissolution of extracellular matrix: contribution of urokinase dependent and metalloproteinase-dependent proteolytic pathways. AB - Constitutive overexpression of both urokinase and matrix metalloproteinase (MMP) activity is frequently observed in individual malignant tumors. In this study we describe the combined contribution of these distinct enzyme systems to the invasive phenotype of a highly metastatic human melanoma cell line (M24met). M24met cells were found to secrete a spectrum of MMPs, including interstitial collagenase, type IV collagenases (M(r) 92,000 and 72,000 progelatinases), and stromelysin. Urokinase, but not tissue-type plasminogen activator, was detected in M24met-conditioned media and on cell surfaces. The contribution of these enzymes to extracellular matrix dissolution was determined by exploiting specific inhibitors, namely tissue inhibitor of the metalloproteinases-2 and plasminogen activator inhibitor-2. Due to the coexpression of urokinase and MMP-dependent activity, M24met cells were observed to degrade multiple components of the extracellular matrix and to significantly degrade both interstitial and basement membrane matrices. Urokinase-dependent removal of matrix glycoprotein was observed to precede MMP-dependent collagenolysis as a prerequisite rate-limiting step. We present evidence which suggests that this temporal relationship is imposed by the structural architecture of the matrix such that matrix glycoprotein serves to protect associated collagen from MMP-dependent degradation. In addition to mediating significant collagenolysis, MMP activity was further implicated in the dissolution of matrix tropoelastin. Urokinase/plasmin activity was not found to be required for MMP-zymogen activation. PMID- 8425206 TI - Adhesion of metastatic, ras-transformed NIH 3T3 cells to osteopontin, fibronectin, and laminin. AB - We previously reported that H-ras-induced metastatic ability in murine NIH 3T3 cells is accompanied by increased expression of osteopontin (OPN). OPN is a secreted phosphoprotein that contains a GRGDS amino acid sequence, suggesting adhesive function, but the function of OPN in tumor cells remains poorly understood. Here we report that PAP2 cells (ras-transformed, metastatic NIH 3T3 cells) adhere and spread on OPN-coated substrates, while NIH 3T3 cells adhere and spread poorly on OPN. A similar pattern was seen for adhesion to laminin, while both cell lines adhered equally well to fibronectin. Adhesive interactions to OPN, laminin, and fibronectin were specific and were blocked by GRGDS (but not control GRGESP) peptides. The kinetics of adhesion to all three substrates was examined. Maximum adhesion was observed at 30-60 min, with reduced adhesion thereafter. We also purified metabolically labeled [32P]OPN secreted by PAP2 cells. Labeled OPN bound better in solution to PAP2 cells than to NIH 3T3 cells, and binding to both cell lines was blocked by GRGDS peptides, results that are consistent with the adhesion and spreading of these cells to OPN-coated substrates. Malignant PAP2 cells thus not only secrete increased levels of OPN, relative to NIH 3T3 cells, but also adhere better to this protein. While the target of OPN secreted by tumor cells is not known, our results raise the possibility that tumor cells that secrete OPN may also bind this protein and that this binding may function in autocrine-type signal transduction important to malignancy. PMID- 8425207 TI - Antitumor effects of recombinant interleukin-6 expressed in eukaryotic cells. AB - In the present study we evaluate the antitumor efficacy of a glycosylated molecule of interleukin-6 (IL-6), which was cloned and expressed in Chinese hamster ovary cells. When tested with two syngeneic murine tumors, the MC38 adenocarcinoma and the MCA106 fibrosarcoma, recombinant IL-6 (rIL-6) significantly reduced the number of day-3 established MC38 lung metastases, but had no effect on MCA106 lung metastases. A similar effect of rIL-6 was seen on day-3 MC38 liver metastases. The antitumor activity mediated by rIL-6 was achieved at doses of the cytokine ranging from 6 micrograms to 150 micrograms/day. There was no correlation between the responsiveness to rIL-6 of these two tumors and their susceptibility, in vitro, to a direct cytostatic effect of the cytokine or the increase in the expression of major histocompatibility complex (MHC) antigens after exposure to rIL-6. However, a correlation was seen between the antitumor response to rIL-6 and the initial number of tumor cells expressing MHC antigens. The possible role of MHC antigens expressed on tumor cells, the generation of MHC-restricted cytotoxic cells and the responsiveness to IL-6 are discussed. PMID- 8425208 TI - Depressed monocyte polarization and clustering of dendritic cells in patients with head and neck cancer: in vitro restoration of this immunosuppression by thymic hormones. AB - The in vitro restoring effects of a thymic hormone preparation, TP-1, on defective monocyte and dendritic cell function in patients with head and neck squamous cell carcinoma (HNSCC) have been examined. The N-formylmethionyl-leucyl phenylalanine (fMLF)-induced polarization of monocytes isolated from the peripheral blood was significantly lower (a mean of 19%) than the polarization of monocytes isolated from healthy controls (a mean of 33%). After the in vitro addition of TP-1 this defective polarization was improved to the normal value of 33% polarized monocytes. The capability of dendritic cells prepared from the blood to form cellular clusters with allogeneic cells was impaired in 26/44 patients. In vitro addition of TP-1 again had restoring effects. The original defective dendritic cell clustering of 97 clusters/six microscopic fields (mean) was improved to a value of 121 clusters. The defects in monocyte polarization and clustering of dendritic cells could be ascribed to the presence in serum of a tumor-derived low-molecular-mass factor low-M(r) factor; < 25 kDa) sharing structural homology with p15E, the capsular protein of murine and feline leukemogenic retroviruses. The incubation of low-M(r) factor from the serum of HNSCC patients with healthy donor monocytes resulted in a significantly higher inhibition of fMLF-induced monocyte polarization than did incubation with control low-M(r) factor (a mean of 42 versus 16% inhibition). This suppressive effect of patient low-M(r) factor was abrogated with a mixture of two monoclonal antibodies against p15E as well as with TP-1. The observations here reported on the in vitro effects of TP-1 on depressed monocyte and dendritic cell function in HNSCC have provided one of the rationales for a TP-1 therapeutic pilot trial recently started in HNSCC patients. PMID- 8425209 TI - Effect of lentinan on macrophage cytotoxicity against metastatic tumor cells. AB - We studied the effect of lentinan, a fungal polysaccharide immunomodulator, on mouse peritoneal macrophages. The i.p. treatment of mice with 10 mg/kg lentinan affected the number, plastic-adherence, and endogen peroxidase activity of peritoneal cells. The cytotoxicity of lentinan-stimulated peritoneal macrophages was determined against several murine and human metastatic tumor targets: Lewis lung carcinoma (LLT) and two human melanomas, and was found to be significantly higher than that of the macrophages from control animals. However, the highly metastatic variant of LLT (LLT-HH) was resistant to the cytolytic effect of resident and lentinan-activated macrophages as well, indicating that the stimulation for cytotoxicity depends not only on the functional activity of the effector but also on the sensitivity of the target. PMID- 8425210 TI - Tumour-cell-induced production of tumour necrosis factor by monocytes of gastric cancer patients receiving BCG immunotherapy. AB - Human peripheral blood monocytes cocultured with tumour cells were used as an in vitro model of in situ interactions between tumour-infiltrating macrophages and the tumour. Tumour cells stimulated de novo expression of the human tumour necrosis factor alpha (TNF) gene in monocytes and caused the release of TNF into the culture supernatant. A group of 14 patients with stage IVA gastric cancer receiving adjuvant chemotherapy (5-FU, Adriamycin, mitomycin C: FAM) or immunochemotherapy (BCG+FAM) was investigated for the ability of monocytes to produce TNF in vitro upon stimulation with tumour cells or purified protein derivative of tuberculin (PPD). Patients were followed at biweekly intervals, i.e. before each instillation of BCG epicutaneously over a period of 10 weeks. It was found that monocytes of some patients receiving BCG at the end of the observation period had an enhanced ability to produce TNF following stimulation with tumour cells. In contrast, such production was not substantially altered during the study period in patients on chemotherapy. PPD-induced TNF production was much weaker and was not significantly changed during this observation time. We infer that BCG immunotherapy may induce the subtle changes in some cancer patients that lead to an increased interaction between monocytes and tumour cells and result in enhanced production of cytokine(s) with antitumour properties. PMID- 8425211 TI - Evaluation of human carcinoembryonic-antigen (CEA)-transduced and non-transduced murine tumors as potential targets for anti-CEA therapies. AB - The MC-38 C57BL/6 mouse colon adenocarcinoma cell line has been transduced with a retroviral construct containing cDNA encoding the human carcinoembryonic antigen (CEA) gene [Robbins PF, Kantor JA, Salgaller M, Horan Hand P, Fernsten PD, Schlom J (1991) Cancer Res 51: 3657]. Two clones, MC-38-ceal and MC-38-cea2, expressed high levels of CEA on their cell surface. A third CEA-expressing cell line, MCA 102-cea3, was similarly derived by transduction of the MCA-102 C57BL/6 mouse fibrosarcoma cell line and is described here. In this study, the three CEA transduced murine tumor cell lines (MC-38-ceal, MC-38-cea2, MCA-102-cea3) were evaluated for their tumorigenic potential, as well as their ability to serve as in vivo model systems for active and passive immunotherapy studies. Parameters that were investigated include tumor growth rate, the antibody response of the host to CEA, and the CEA content of the tumors. The MC-38-cea2 model appeared to be the most appropriate for immunotherapy studies. Biodistribution studies, using an 125I-labeled anti-CEA mAb, demonstrated efficient tumor targeting of MC-38 cea2 tumors in C57BL/6 and athymic mice. PMID- 8425212 TI - Recognition of the Qa-2k tumor antigen by T cell receptor gamma/delta of an immunopotentiator-induced tumoricidal T cell of mice. AB - Tumor-specific expression of Qa-2k antigen coded by the Q5k gene on various mouse tumor cells and immunological response of the host mice to the antigen have been demonstrated [Seo et al. (1992) J Exp Med 175: 547; Tanino et al. (1992) Cancer Immunol Immunother 35: 230]. The possibility was examined that Qa-2 antigen is one of the recognition target molecules of immunopotentiator-induced, H-2 nonrestricted tumoricidal lymphocytes of Qa-2-mice. Lymphocytes stimulated in vivo with P. acnes or culture-induced anomalous killers of B6.K1 mice did not exhibit significant in vitro cytotoxicity against B6.K1 lymphoblasts but lysed their Qa-2,3-congenic counterpart B6 lymphoblasts. To demonstrate the Qa-2 specificity of such cytotoxic cells more precisely, an L cell transformant clone (LQ7b/Kb), which expressed the alpha 1 and alpha 2 domains of the Qa-2 antigen (Q7b gene product), was generated by transfecting a cloned plasmid DNA containing a hybrid gene constructed from the 5' half of the Q7b gene and the 3' half of the H-2Kb gene (pQ7b/Kb). Using LQ7b/Kb cells as the target cells and the nylon-wool nonadherent fraction of lymphocytes from P. acnes-stimulated (C3H/He x B6.K1)F1 mice (H-2k, Qa-2-) as the effector cells of the in vitro cytotoxicity reaction, the presence of cytotoxic cells that recognize the alpha 1/alpha 2 region of the Q7b gene product was demonstrated. The cytotoxic activity was dependent on T cells bearing T cell receptors of the gamma/delta type (TCR gamma/delta). The (C3H/He x B6.K1)F1 effector cells, as well as the B6.K1 effector cells also lysed BW5147 lymphoma cells (Qa-2k+) derived from AKR mice (Qa-2-, H-2k). By target competition experiments it was shown that some of the effector cells lytic to BW5147 were identical to those that lysed LQ7b/Kb. Therefore some of the tumoricidal cells induced by the immunopotentiator interact with the target tumor cells through recognition of the alpha 1/alpha 2 region of the Qa-2k tumor antigen by TCR gamma/delta. PMID- 8425213 TI - Superantigen-based tumor therapy: in vivo activation of cytotoxic T cells. AB - We have recently demonstrated that the superantigen staphylococcal enterotoxin A (SEA) targets in vitro activated cytotoxic T lymphocytes against tumor cells expressing major histocompatibility complex (MHC) class II antigens. In this report we analyze the use of SEA as an immunoactivator in vivo. Treatment of mice with SEA activated a fraction of CD3+ T cells apparently as a function of their T cell receptor V beta expression. SEA induced interleukin-2 receptor expression and proliferation in both CD4+ and CD8+ T cells. This proliferative response was dose-dependent (0.1-100 micrograms/mouse), peaked during day 1 after treatment and declined to background levels within 4 days. The cytotoxic response, measured as cytotoxicity to SEA-coated MHC class II+ target cells (staphylococcal enterotoxin-dependent cell-mediated cytotoxicity, SDCC), was maximal at a dosage of 1 microgram SEA/mouse. The SDCC was confined to the CD8+ T cell compartment, peaked 2 days after treatment and declined to background levels within 4 days. A second injection of SEA on day 5 after the first SEA treatment resulted in SDCC function with kinetics and magnitude identical to that seen after one injection. These results pave the way for the use of SEA in the treatment of MHC class II+ tumors. PMID- 8425214 TI - Characterization of a monoclonal antibody reactive with a glycolipid antigen expressed by tumorigenic and certain immortalized, non-tumorigenic rat esophageal epithelial cell lines. AB - A monoclonal antibody (mAb 5G) was produced against a tumorigenic rat esophageal epithelial cell line, designated B2T. Using an enzyme-linked immunosorbent assay, immunofluorescence assay (IFA), thin-layer chromatography (TLC) and immunoperoxidase staining, it was found that mAb 5G reacted specifically with a glycolipid antigen expressed by three tumorigenic rat esophageal epithelial cell lines, and two out of the three non-tumorigenic, immortalized rat esophageal epithelial cell lines tested; but did not react with primary cultures of normal rat esophageal epithelial cells or fibroblasts. mAb 5G did not bind to rat respiratory tract carcinoma cell lines, to immortalized rat tracheal epithelial cell lines, or to primary cultures of normal rat tracheal epithelial cells. In addition, mAb 5G did not react with any of the human or mouse cell lines tested. In IFA experiments, mAb 5G stained imprints prepared from in vivo propagated B2T tumor tissues, but did not react with normal rat esophageal, tracheal, lung, liver, and kidney tissues. The antigen was identified by TLC as a neutral glycolipid, consisting of two bands, with RF = 0.45 and 0.41, which migrated in proximity to the ceramide trihexoside standard on TLC plates. Densitometric scanning of the antigen bands indicated that the tumorigenic rat esophageal cell lines possessed 50%-90% more mAb-5G-reactive antigen than the non-tumorigenic esophageal cell lines. The results show that mAb 5G reacts specifically with a glycolipid antigen expressed by tumorigenic and certain non-tumorigenic, immortalized rat esophageal epithelial cell lines that might be at the late stages of transformation and early malignancy. PMID- 8425215 TI - mRNA splicing and autocatalytic introns: distant cousins or the products of chemical determinism? PMID- 8425216 TI - Survival of hunger and stress: the role of rpoS in early stationary phase gene regulation in E. coli. PMID- 8425217 TI - Expression of Xist during mouse development suggests a role in the initiation of X chromosome inactivation. AB - The mouse Xist gene maps to the X inactivation center (Xic) region and is expressed exclusively from the inactive X chromosome. It is thus a candidate gene for the Xic. We show that the onset of Xist expression in mouse development precedes X chromosome inactivation and may therefore be a cause rather than merely a consequence of X inactivation. The earliest Xist expression in morulae and blastocysts is imprinted, resulting in specific expression of the paternal Xist allele. Imprinted Xist expression may thus be the cause of nonrandom inactivation of the paternal X in trophectoderm. Strong Xce alleles can act to reduce the effect of imprinted Xist expression in the trophectoderm. The imprint on Xist expression is lost shortly before gastrulation when random X inactivation occurs. Our data support a direct role for Xist in the initiation of X inactivation. PMID- 8425218 TI - Mad: a heterodimeric partner for Max that antagonizes Myc transcriptional activity. AB - Myc family proteins appear to function through heterodimerization with the stable, constitutively expressed bHLH-Zip protein, Max. To determine whether Max mediates the function of regulatory proteins other than Myc, we screened a lambda gt11 expression library with radiolabeled Max protein. One cDNA identified encodes a new member of the bHLH-Zip protein family, Mad. Human Mad protein homodimerizes poorly but binds Max in vitro, forming a sequence-specific DNA binding complex with properties very similar to those of Myc-Max. Both Myc-Max and Mad-Max heterocomplexes are favored over Max homodimers, and, unlike Max homodimers, the DNA binding activity of the heterodimers is unaffected by CKII phosphorylation. Mad does not associate with Myc or with representative bHLH, bZip, or bHLH-Zip proteins. In vivo transactivation assays suggest that Myc-Max and Mad-Max complexes have opposing functions in transcription and that Max plays a central role in this network of transcription factors. PMID- 8425219 TI - Mxi1, a protein that specifically interacts with Max to bind Myc-Max recognition sites. AB - We used the interaction trap to isolate a novel human protein that specifically interacts with Max. This protein, Mxi1 (for Max interactor 1), contains a bHLH Zip motif that is similar to that found in Myc family proteins. Mxi1 interacts specifically with Max to form heterodimers that efficiently bind to the Myc-Max consensus recognition site. When bound to DNA by a LexA moiety in yeast, Mxi1 does not stimulate transcription. mxi1 mRNA is expressed in many tissues, and its expression is elevated in U-937 myeloid leukemia cells that have been stimulated to differentiate. These facts are consistent with a model in which Mxi1-Max heterodimers indirectly inhibit Myc function in two ways: first, by sequestering Max, thus preventing the formation of Myc-Max heterodimers, and second, by competing with Myc-Max heterodimers for binding to target sites. PMID- 8425220 TI - Oncogenic activity of the c-Myc protein requires dimerization with Max. AB - c-Myc (Myc) and Max proteins dimerize and bind DNA through basic-helix-loop-helix leucine zipper motifs (b-HLH-LZ). Using a genetic approach, we demonstrate that binding to Max is essential for Myc transforming activity and that Myc homodimers are inactive. Mutants of Myc and Max that bind efficiently to each other but not to their wild-type partners were generated by either exchanging the HLH-LZ domains or reciprocally modifying LZ dimerization specificities. While transformation defective on their own, complementary mutants restore Myc transforming activity when coexpressed in cells. The HLH-LZ exchange mutants also have dominant negative activity on wild-type Myc function. In addition, wild-type max antagonizes myc function in a dose-dependent manner, presumably through competition of Max-Max and Myc-Max dimers for common target DNA sites. Therefore, Max can function as both suppressor and activator of Myc. A general model for the role of Myc and Max in growth control is discussed. PMID- 8425222 TI - Characterization of murine lymphokine-activated killer cell cultures separated according to cell size. AB - Incubation of murine splenocytes in high concentrations of the lymphokine interleukin 2 leads to the development of cytotoxic effector cells termed lymphokine-activated killer cells. Two populations of blast cells develop within these cultures, one of T origin and one of NK origin. The T blasts are CD8+, FcR gamma III- and exhibit slightly increased forward angle light scatter (FSC) and side angle light scatter (SSC). They have little or no oncolytic activity against the tumor target YAC-1. The NK blasts are CD8-, FcR gamma III+ and exhibit markedly increased FSC and SSC. They contain virtually all the lytic activity against the tumor target YAC-1. Cultures were split into multiple fractions on the basis of sedimentation velocity, which separates cells primarily on the basis of cell size. The sedimentation separation resolved both T and NK cells into resting (G0) small lymphocytes and cycling blast cells. NK blasts sedimented much more rapidly than all CD8+ cells such that most of the oncolytic activity could be separated relatively free of CD8+ cells. PMID- 8425221 TI - Deficient expression of a B cell cytoplasmic tyrosine kinase in human X-linked agammaglobulinemia. AB - We describe a novel cytoplasmic tyrosine kinase, termed BPK (B cell progenitor kinase), which is expressed in all stages of the B lineage and in myeloid cells. BPK has classic SH1, SH2, and SH3 domains, but lacks myristylation signals and a regulatory phosphorylation site corresponding to tyrosine 527 of c-src. BPK has a long, basic amino-terminal region upstream of the SH3 domain. BPK was evaluated as a candidate for human X-linked agammaglobulinemia (XLA), an inherited immunodeficiency characterized by a severe deficit of B and plasma cells and profound hypogammaglobulinemia. BPK mapped to within 100 kb of a probe defining the polymorphism most closely linked to XLA at DXS178. Reduction in or the absence of BPK mRNA, protein expression, and kinase activity was observed in XLA pre-B and B cell lines. BPK is likely the XLA gene and functions in pathways critical to B cell expansion. PMID- 8425223 TI - T cell-B cell interaction: autoreactive T cells recognize B cells through a terminal mannose-containing superantigen-like glycoprotein. AB - Rat autoreactive T cells (ATs) recognize a membrane component(s) on syngeneic B cells in association with class II MHC antigens resulting in proliferation of ATs as well activation and differentiation of B cells. Results presented herein indicate that ATs recognize a stimulating antigen(s) SA, in association with class II MHC antigens, on the B cell surface. Our studies using inhibitors of carbohydrate and protein synthesis suggest that SA is a glycoprotein(s) with a high turnover rate but is not an immunoglobulin. Treatment of B cells with mannosidase abrogates their ability to stimulate AT proliferation. Furthermore, pretreatment of B cells with GNA (a lectin from Galanthus nivalis that reacts with free terminal-mannose residues on glycoconjugates) also inhibits their ability to stimulate ATs. However, these treatments do not affect the competence of B cells to stimulate an allogeneic MLR or present a conventional antigen to T cells. The frequency of CD4+ T cells proliferating in response to syngeneic B cells is very high (0.2-0.5%) and is in line with frequencies seen in "superantigen"-type responses. Moreover, T cell receptors expressed on ATs use mainly V beta 6, V beta 11, and V beta 8 regions. Based on these data, SA appears to be a fast turnover, terminal mannose-containing, superantigen-like glycoprotein on the B cell surface. PMID- 8425224 TI - Pristane induces an indomethacin inhibitable inflammatory influx of CD4+ T cells and IFN-gamma production in plasmacytoma-susceptible BALB/cAnPt mice. AB - In BALB/cAnPt (BALB/c) mice, the intraperitoneal injection of pristane induces the formation of chronic oil granulomatous tissue and a high incidence of plasmacytomas (PCT). DBA/2N (DBA) and (BALB/c x DBA)F1 hybrid (CDF1) mice develop oil granulomas but no PCT. Recent studies comparing pristane-injected conventionally housed BALB/c mice with specific pathogen-free BALB/c mice (SPF) demonstrated a significant reduction in both the incidence of PCT and the T cell infiltration of oil granulomatous tissue in SPF mice. In this study, FACS analysis was performed to determine the proportion of myeloid, T. and B cells present in pristane-induced peritoneal exudate (PE), and oil granulomas (OG) of BALB/c, DBA, and CDF1 mice. At all time points studied, the majority of cells recovered from the PE and OG of all three strains of mice were Mac-1+, presumably macrophages and neutrophils. Neither Ly-5(B220)+ B cells nor CD8+ T cells were significantly altered by pristane injection. BALB/c mice had a dramatic influx of CD4+ T lymphocytes (three- to fivefold) more than 50 days after the initial injection of pristane. The PCT-resistant DBA and CDF1 mice did not. This increase in CD4+ cells in BALB/c mice was not significantly affected by a second injection of pristane nor was it induced by a second injection in DBA mice. Indomethacin, which has been shown to prevent the development of PCT in BALB/c mice, prevented the infiltration of CD4+ T cells. In addition, pristane-induced levels of interferon-gamma greater than controls were found in the peritoneal lavages of BALB/c mice at all time points tested but not in DBA or indomethacin-treated BALB/c mice. In contrast, pristane injection increased levels of interleukin-5 in DBA but not BALB/c mice. PMID- 8425225 TI - Interleukin-4 signals regulating CD23 gene expression in human B cells: protein kinase C-independent signaling pathways. AB - Signal transduction by IL-4 leading to the activation of CD23(Fc epsilon RII) gene expression using human tonsillar B cells was studied. IL-4 stimulated CD23 mRNA transcription within hours (1-4 hr) which preceded the later induction of cell surface CD23. The induction of CD23 gene transcription by IL-4 was not adversely affected by cycloheximide, suggesting that post-translational modifications are accounted for the gene activation. PKC activators (PMA, diacylglycerol, indolactam) were effective inducers of CD23 gene expression, whereas calcium ionophores were not. PMA and IL-4 also displayed similar induction kinetics for CD23 mRNA. However, the signaling pathways utilized by the two agents appear distinct as shown by (1) cotreatment of IL-4 and PMA caused CD23 gene expression over the maximum level inducible by each agent alone and (2) unlike the PMA-induced CD23 expression, the IL-4-induced expression was not affected by PKC inhibitors. These results strongly suggest that IL-4 signals leading to CD23 gene activation are mediated via a PKC-independent pathway. A possible role of tyrosine kinases in the regulation of CD23 expression is discussed. PMID- 8425226 TI - Human immune response to cationized proteins. I. Characterization of the in vitro response to cationized diphtheria toxoid. AB - Cationization of proteins, i.e., increasing net positive charge by the substitution of carboxyl groups with positively charged residues, has been reported to enhance protein immunogenicity in animal model systems. In the present study, we have investigated the effect of cationization on the in vitro cell-mediated immune response of human mononuclear cells to diphtheria toxoid. A series of cationized DT preparations were generated by covalent modification with ethylenediamine, with pIs ranging from 4.6 to > 9.3, and tested for their ability to induce proliferation of normal human peripheral blood mononuclear cells. Cationized DT (cDT) was found to induce an antigen-specific, augmented proliferative response, relative to native antigen, which was directly proportional to the degree of cationization. Further characterization of the response to cDT demonstrated that (1) proliferative responses could be detected considerably earlier, and typically at much lower antigen concentrations, than the response to native DT; (2) the response was dependent on HLA-DR; (3) production of a number of cytokines, sp. IL-1 beta, IL-2, and IFN-gamma, was also elevated in cDT-stimulated cultures; and (4) the enhanced proliferative response to cDT could be attributed to CD4+ helper T cells. These results demonstrate that cationization of proteins enhances the ability to generate a cell-mediated immune response in humans and suggest that cationization may have utility in the design of more effective carrier proteins for human vaccines. PMID- 8425227 TI - Human immune response to cationized proteins. II. Characterization of interaction of cationized diphtheria toxoid with human mononuclear cells. AB - Cationized diphtheria toxoid (cDT) has previously been shown to be more effective than the native protein as an inducer of human antigen-specific T cell responses. In the present study, biotin-labeled antigen and flow cytometric analysis were used to examine the possibility that enhanced immunogenicity of cDT may be a consequence of preferential binding to antigen-presenting cells. Strong binding of cDT, relative to native antigen, was noted for both monocytes and B cells. Characteristics of binding were similar for both cell types, including rapid saturation, temperature independence, and inhibition by unlabeled cationized proteins. Although both B cells and monocytes bound cDT, only monocytes were effective in triggering T cell proliferation, possibly as a result of slow internalization of bound antigen by B cells. Definition of the target structures of cationized proteins may allow for the design of more efficient vaccines, which would be specifically targeted to antigen-presenting cells in vivo. PMID- 8425228 TI - Biphasic effect of kallikrein on IgE and IgG1 syntheses by LPS/IL-4-stimulated B cells. AB - epsilon receptor modulating protein (epsilon RMP) was identified and purified in our previous studies as a murine T cell-derived soluble 17-kDa chymotryptic serine protease which suppresses avidity of binding between IgE and CD23 (low affinity Fc receptor for IgE) without decreasing the quantitative expression of the CD23 molecule. Some, but not all, of the other known soluble serine proteases showed epsilon RMP-like CD23-modulating activities. Further studies indicated that epsilon RMP exists not only as a soluble protein but also as a 36-kDa T-cell surface form. Both soluble and membrane-bound epsilon RMP can induce purified splenic B cells to secrete IgE in the presence of IL-4 even without lipopolysaccharide (LPS). In this study, therefore, we have tested effects of several known serine proteases on Ig production in vitro and have found that: (i) coculture of splenic B cells in the presence of LPS and IL-4 with serine proteases which have epsilon RMP-like substrate specificity, such as kallikrein and alpha-chymotrypsin, results in a significant increase of IgG1 and a slight increase of IgE secretion at low concentrations, and significant suppression at high concentrations in an isotype-selective manner; and (ii) the effects of these proteases are blocked by phenylmethylsulfonyl fluoride but not by indomethacin, suggesting that serine protease activity but not prostaglandin E2 is involved. The biological significance of the possible involvement of serine proteases on Ig class switching is discussed. PMID- 8425229 TI - Microtubules are not an essential component of phytohemagglutinin-dependent signal transduction in Jurkat T lymphocytes. AB - We have used two disruptors of the cytoskeleton microtubular network (colchicine and vinblastine) to investigate the role of microtubules in Phaseolus vulgaris phytohemagglutinin (PHA) signal transduction in Jurkat T cells. Both drugs decreased but did not abolish the PHA-dependent Ca2+ response of Jurkat cells. Neither colchicine nor vinblastine had any major effects on the PHA-dependent turnover of the mono-, di-, tri-, and tetra-substituted phosphorylated derivatives of D-myo-inositol or the cellular distribution of protein kinase C (PKC) activity. However, both microtubule disruptors increased interleukin-2 (IL 2) production. Whereas vinblastine enhanced IL-2 production approximately twofold at all the concentrations tested (0.1, 1.0, and 10 microM), colchicine did so only at a 10 microM concentration. When a combination of PHA and 12-O tetradecanoyl-13-O-acetyl phorbol (TPA) was used, a small increase in IL-2 production was observed only in the presence of vinblastine (10 microM). In contrast to recent reports that microfilaments may be involved in the regulation of signal transduction, our data suggest that this is not the case for microtubules in PHA-dependent signal transduction in Jurkat T cells. PMID- 8425230 TI - Interleukin-2 induces apoptosis in mouse thymocytes. AB - Interleukins play a role in the process of T-cell development and, like other cytokines, seem able to modulate apoptosis. Interleukin-2 has been reported to inhibit apoptotic cell death of thymocytes induced in vitro by either activation of CD3/TCR complex or treatment with glucocorticoid hormone. We demonstrate here that IL-2 can provoke DNA fragmentation and cell death of CD4+ CD8+ mouse thymocytes by activating an endogenous apoptotic pathway. Thymocytes, incubated with high IL-2 concentrations in vitro, showed the morphological characteristics of apoptotic cells, including reduction in nuclear size, derangement in chromatin structure, and DNA fragmentation in oligonucleosomal subunits. Inhibition of mRNA and protein synthesis and addition of the PKC-inhibitor H-7, Zn2+ ions, and IL-4 counteracted the IL-2 effect. These data suggest that high IL-2 concentrations may induce an active process of cell death on mouse thymocytes in vitro. PMID- 8425231 TI - The effect of T cell-derived cytokines on B cell motility in vitro. AB - We have investigated the ability of interleukin-1 (IL-1), IL-2, IL-3, IL-4, IL-5, IL-6, and interferon-gamma (IFN-gamma) to induce motility in murine, splenic B lymphocytes. Two parameters of cellular locomotion were studied, namely acquisition of motile morphology (polarization) and in vitro migration through polycarbonate filters. Of the tested cytokines, only IL-4 gave a strong motile response among B cells. At the optimal concentration of 3 ng/ml, IL-4 induced polarization in 10-20% of the B cells within 30 min and in up to 40% after overnight exposure. The same amount IL-4 present in a microchemotaxis chamber stimulated migration through polycarbonate filters after overnight incubation in up to 15% of the input B cells. IFN-gamma could also induce some degree of polarization and migration, but only after 19 hr of exposure and to a much lesser extent than IL-4. PMID- 8425232 TI - Activated synovial T cell clones from a patient with rheumatoid arthritis induce proliferation of autologous peripheral blood-derived T cells. AB - In order to investigate cellular interactions involved in the development of human autoimmune disease, a synovial fluid-derived T cell clone reactive with mycobacterial antigens, termed k38, was employed as a stimulus for autologous peripheral blood mononuclear cells (PBMC). Stimulator cells were used either activated with immobilized OKT3 mAb or in a resting state. Activated k38 cells triggered PBMC to proliferate. A T cell line prepared by coculturing autologous PBMC with irradiated activated k38 cells proliferated upon stimulation with activated k38 cells in the presence of PBMC as a source of accessory cells, as did T cell clones that were subsequently isolated from this line. Blocking studies revealed that proliferation of the anti-k38 line and anti-k38 clones in response to stimulation with clone k38 could be inhibited by monoclonal antibodies against a variety of cellular determinants including HLA class I and LFA-1 beta. It was demonstrated that the antigen reactivity of clone k38 was modulated by the presence of anti-k38 clones. These data provide a model for understanding the cellular interactions that may take place in vivo in the evolution of the chronic synovial inflammatory process. PMID- 8425233 TI - Myelin-liposome protection against experimental autoimmune encephalomyelitis is associated with reduced neuroantigen-specific T-cell-mediated responses. AB - Lewis rats undergo a relapsing paralytic disease upon challenge with spinal cord emulsified in complete Freund's adjuvant (CFA). Treatment with two intracardiac injections of liposomes composed of whole myelin significantly reduced the severity of disease. Protection was disease-specific since treatment with myelin liposomes did not protect Lewis rats against adjuvant arthritis (AA), a CNS unrelated T-cell-mediated autoimmune disease. Myelin-liposome-treated, spinal cord/CFA-immunized rats displayed borderline reduction of delayed-type hypersensitivity (DTH) (ear swelling) reactions to myelin and myelin basic protein (MBP), but significantly reduced in vitro lymphnode cell proliferation in response to these antigens. Responses to purified protein derivative of Mycobacterium tuberculosis (PPD) were not reduced, emphasizing the antigen specific nature of the myelin-liposome-mediated suppression. Spleen cell proliferative responses were inconsistent and often poor. However, when cultured in the presence of NG-monomethyl-L-arginine (MMA), antigen-specific proliferation of spleen cells from both treated and control rats was greatly enhanced, indicating that reactive nitrogen intermediates contributed to the decrease in spleen cell proliferation. Purified splenic T cells from treated rats displayed a pattern of proliferation similar to that of unseparated lymphnode cells. Treatment of rats with a single injection of myelin liposomes after recovery from the first clinical episode significantly reduced the severity of the relapses. PMID- 8425234 TI - T cell lymphokine-induced secretion of cytokines by monocytes from patients with multiple sclerosis. AB - To investigate the function of peripheral blood monocytes in multiple sclerosis (MS), we measured the production of the cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha (TNF alpha), and interleukin-6 (IL-6), and the procoagulant, tissue factor (TF) in 17 patients with chronic progressive MS and 15 normal controls. Monocyte activity was tested under unstimulated, minimal endotoxin conditions and after culture with various stimuli, including Escherichia coli lipopolysaccharide (LPS), crude supernatant from anti-CD3 activated T cells, recombinant interleukin-2 (rIL-2), and recombinant interferon gamma (rIFN-gamma). A higher number of MS patients than controls had circulating monocytes which spontaneously secreted IL-6 or contained detectable cell associated IL-1 beta. Monocyte responses to LPS were comparable between the two groups; LPS caused production and secretion of all cytokines and TF in every MS patient and control. In contrast, crude T cell supernatants, rIL-2, and rIFN gamma induced IL-1 beta release in a higher number of MS monocytes than that in controls, whereas the production and secretion of the other cytokines and TF activity were similar between the groups. We conclude that some MS patients have "primed" circulating monocytes, as shown by excessive spontaneous IL-6 release and intracellular IL-1 beta synthesis. Unstimulated MS monocytes, however, are not different from controls with respect to spontaneous secretion of small amounts of IL-1 beta and TNF alpha and expression of cell surface TF. Excessive IL-1 beta secretion by MS monocytes after stimulation with T-cell-derived lymphokines suggests dysregulation of T cell-monocyte interactions which may be most relevant in the central nervous system plaques where activated T cells are found. PMID- 8425235 TI - Vaporizer overfilling. PMID- 8425236 TI - Flumazenil reduces the duration of thiopentone but not of propofol anaesthesia in humans. AB - The effect of flumazenil (F) on the duration of anaesthesia produced by a single dose of thiopentone (T) and propofol (P) was investigated in a placebo-controlled double-blind trial. Eighty-four patients anaesthetized with N2O in O2 and either thiopentone 7 mg.kg-1 or propofol 3 mg.kg-1 for minor gynaecological procedures were studied. Patients were randomly allocated to pretreatment with either 0.5 mg of flumazenil (F) or 5 ml of normal saline (NS) in one of the following groups: T/NS, T/F, P/NS, or P/F. Anaesthetic requirements were assessed by recording the time between the injection of anaesthetic and the first movement observed during the procedure. The time elapsed from the administration of thiopentone to the first movement was 6.5 +/- 1.6 min for the T/NS group and 5.3 +/- 2.4 min for the T/F group (P < 0.05). The first movement after propofol administration was observed at 7.0 +/- 2.2 min in the P/NS group and at 7.1 +/- 4.5 min in the P/F group (NS). These data suggest that pretreatment with 0.5 mg of flumazenil iv reduces the duration of thiopentone but not of propofol anaesthesia. PMID- 8425237 TI - Comparison between alprazolam and hydroxyzine for oral premedication. AB - The safety and efficacy of alprazolam and hydroxyzine administered orally as surgical premedicants were compared in a double-blind controlled study. Sixty five patients were given either alprazolam 1 mg or hydroxyzine 75 mg, one to two hours before surgery. Anxiety was assessed by both the patient and the anaesthetist, the patient using a visual analogue scale, the anaesthetist employing both analogue and ordinal ratings. Sedation was assessed by the anaesthetist only, using the same two methods. Amnesia was appraised with a simple memory test. Safety was assessed by recording adverse effects and measuring haemodynamic variables. Premedication with alprazolam produced a modest reduction in anxiety (28%) (P < 0.01) while hydroxyzine had no detectable effect. The comparison of the sedation level and of the memory test revealed no difference between the two premedicants. Minor side effects were only observed in the hydroxyzine group. Changes in blood pressure were more pronounced in the hydroxyzine group. This study shows that alprazolam and hydroxyzine are safe and efficient oral premedicants. However, alprazolam is preferable to hydroxyzine in terms of anxiolytic and adverse effects. PMID- 8425238 TI - Halothane concentrations required to block the cardiovascular responses to incision (MAC CVR) in infants and children. AB - The purpose of this study was to determine the halothane concentration in N2O required to block the cardiovascular responses to skin incision (MAC CVR) in infants and children. We studied 64 unpremedicated ASA 1 infants and children (one month to seven years). In each infant or child, anaesthesia was induced slowly with halothane and N2O, and an endotracheal tube was placed. The MAC CVR was assessed, after a steady state end-tidal halothane concentration had been established for ten minutes, by the "up and down technique" of Dixon. Positive responses were defined as an increase in MAP or HR > 10%. The MAC CVR50 values of halothane with 60% N2O were 1.16 +/- 0.23% at 1-6 mo, 1.17 +/- 0.18% at 7-12 mo, 0.95 +/- 0.26% at 1-3 yr, and 1.12 +/- 0.16% at 4-7 yr. The value at 1-3 years children was less than those in the other age groups (P < 0.05). The changes of MAP were correlated with changes of both HR and pupillary diameter. These results indicate that the values of MAC CVR50 of halothane in infants and children are higher than those required to block motor responses (MAC). The halothane requirement to block cardiovascular responses is lowest in the children aged one to three years. PMID- 8425239 TI - Use of a tourniquet in patients with sickle-cell disease. AB - Fifteen patients, 13 male and two female, known to be carrying the sickle-cell gene (12 HbSS and 3 HbAS), who were undergoing operations requiring a bloodless field, were included in the study. Of the 12 with HbSS, seven had haemoglobin A1 component of between 11 and 27%, three had fetal haemoglobin ranging from 5.7 to 29% and the remaining two had increased haemoglobin A2 concentrations suggesting a beta non-thalassaemia combination. All had a tourniquet applied to the appropriate limb and were given general anaesthesia with moderate hyperventilation throughout the procedure. The tourniquet inflation time was 61.7 +/- 27.5 min. The mean PaO2 remained above 200 mmHg, mean PaCO2 was less than 37 mmHg, and pH ranged between 7.40 and 7.45. There were no clinically important changes in BP or ECG. All patients made uneventful recoveries and none developed sickle-cell crises. It is suggested that it is safe to use tourniquet in patients with sickle-cell disease provided optimum acid-base status and oxygenation are maintained throughout the procedure. PMID- 8425240 TI - The initial distribution volume of glucose and cardiac output in the critically ill. AB - Blood or plasma glucose concentration can be measured accurately and rapidly. However, after a glucose challenge metabolism may modify glucose kinetics, so that glucose has not been used as an indicator for dilution volumetry. To test the hypothesis that the initial distribution volume of glucose (IDVG) reflects cardiac output rather than glucose metabolism in the critically ill, the relationship between IDVG and thermodilution cardiac output was evaluated at 27 points in 13 non-surgical, critically ill patients without congestive heart failure. The IDVG was calculated from incremental plasma glucose concentrations using a one compartment model. Correlations were obtained between the IDVG and cardiac output (r = 0.89, n = 27, P < 0.001), and between the incremental plasma glucose concentrations three minutes after the injection and the IDVG (r = 0.94, n = 27, P < 0.001). No difference was found between the IDVG with or without continuous insulin infusions. The results indicate that the IDVG reflects cardiac output rather than glucose metabolism in patients without congestive heart failure. PMID- 8425241 TI - Blood sugar, serum insulin and serum non-esterified fatty acid levels during thiopentone anaesthesia in dogs. AB - The purpose of this study was to determine the effect of thiopentone anaesthesia on glucose metabolism. Blood sugar (BS), serum immunoreactive insulin (IRI) and serum non-esterified fatty acid (NEFA) concentrations were measured during the course of (1) an intravenous glucose tolerance test (IVGTT), and (2) an intravenous insulin test (ITT), in conscious and anaesthetized fasted dogs. The IVGTTs were repeated in dogs under alpha- or beta-adrenergic blockade, induced by phentolamine or propranolol. During the IVGTT, the anaesthetized dogs showed glucose intolerance (blood sugar levels were higher than in the control group) and little serum IRI response to hyperglycaemia was detected. An attenuated initial decrease and a slower rebound of NEFA concentration was observed in anaesthetized animals than in controls. Phentolamine administration (5 mg.kg-1 iv) partly restored the IRI response without affecting the BS levels; propanolol (1 mg.kg-1 iv) had no effect. Anaesthetized dogs showed a moderate resistance to insulin induced hypoglycaemic action and a lack of serum NEFA response during counter-regulation of hypoglycaemia, while in conscious controls an intense rebound was observed. Hyperinsulinaemia after iv insulin administration was longer in anaesthetized dogs than in controls. The insulin distribution space was 78% of body weight and insulin t1/2 in blood group compared with 54% and 16 min, in controls. We conclude that thiopentone provokes disturbances in glucose and serum NEFA metabolisms and abolishes the serum IRI response to hyperglycaemia. These effects are influenced by extrapancreatic factors regulating serum IRI levels and by an alpha-adrenergic mechanism, via the inhibition of insulin secretion. PMID- 8425242 TI - Does ranitidine provide protection against acid gastroesophageal reflux? AB - Continuous gastroesophageal pH monitoring was used to evaluate the effect of ranitidine on gastroesophageal reflux (GOR) in 60 patients scheduled for elective non-gastrointestinal abdominal or gynaecological surgery. The patients were randomly assigned to receive a single dose of ranitidine 50 mg either iv (RANIV group) or im (RANIM group) or a placebo iv (PLAC group) 90 min before surgery. The pH was measured continuously for six hours in the lower oesophagus using a flexible calibrated glass electrode. A pH < 4.0 was chosen as the boundary for defining occurrence of acid GOR. Both ranitidine treatments reduced the total number of acid reflux episodes and the global reflux index (P < 0.05). The duration of the acid reflux episodes (sum of refluxes) and the number of acid reflux episodes longer than five minutes were markedly decreased by ranitidine but the mean duration of the reflux episodes was unaffected. The pH values at induction, intubation, surgical incision and extubation were similar in the PLAC and RANIV groups but more acid than in the RANIM group. The mean pH of reflux episodes was less acid in both ranitidine groups than in the PLAC group (P < 0.05). Also the number of very acid refluxes (pH < 2.5) decreased with ranitidine (P < 0.05). Intramuscular and intravenous administration of ranitidine provide protection against gastroesophageal reflux, with shorter duration of reflux episodes occurring in the intramuscular group. Regardless of the administration of ranitidine, protection against gastroesophageal reflux is incomplete; the frequency of reflux episodes is reduced but not eliminated. PMID- 8425243 TI - Anterior mediastinal mass in a patient susceptible to malignant hyperthermia. AB - We report a malignant hyperthermia-susceptible patient who required investigation for a large, symptomatic anterior mediastinal mass. Multiple attempts at tissue diagnosis under local anaesthesia were unsuccessful. Following awake fibreoptic tracheal intubation, general anaesthesia was administered using ketamine, midazolam, and nitrous oxide, maintaining spontaneous ventilation. Prophylactic dantrolene was not used, to avoid potential muscle weakness and respiratory compromise. Diagnostic mediastinotomy was performed without incident. We conclude that ketamine anaesthesia is appropriate for patients with anterior mediastinal masses, and is considered safe in malignant hyperthermia-susceptible patients. PMID- 8425244 TI - Diffuse idiopathic skeletal hyperostosis: an unusual cause of difficult intubation. AB - A case is reported in which anterior osteophytes on the cervical vertebra, in combination with a subglottic stenosis, resulted in distortion of the airway and led to unexpected difficulties during intubation. The osteophytes, associated with the syndrome of diffuse idiopathic skeletal hyperostosis (DISH) were centred at the midcervical level and resulted in anterior displacement of the larynx with an acute angulation of the trachea just below the larynx. This acute angulation, immediately above an unrecognized subglottic stenosis, rendered it impossible to pass all but the smallest endotracheal tube. Diffuse idiopathic skeletal hyperostosis is an ossifying diathesis leading to bone formation in spinal and extraspinal sites, paravertebral osteophyte formation and ligamentous calcification and ossification. Ossification of the anterior longitudinal ligament is common, may be discontinuous, and is often more marked in the thoracolumbar spine than elsewhere. However, isolated and predominant cervical spinal involvement may occur. Diffuse idiopathic skeletal hyperostosis occurs primarily in the elderly population and is often associated with the syndromes of osteoarthritis and ossification of the posterior longitudinal ligament (OPLL). Difficult intubation resulting from anatomical abnormalities of the cervical spine is rare. Although radiological evaluation may be useful in assessing the airway in patients deemed to be at risk for difficult intubation, it cannot be recommended for screening patient populations on a routine basis because of the cost and anticipated extremely low yield. Careful clinical evaluation of the airway before operation and having an approach to the unexpected difficult intubation are emphasized. PMID- 8425245 TI - Carinal resection with two high-frequency jet ventilation delivery systems. AB - A 76-yr-old man underwent carinal resection for squamous cell carcinoma through the right posterolateral thoracotomy approach. Ventilation was maintained by the use of two high-frequency jet ventilators, each attached to a separate catheter during the time of resection and reconstruction of the tracheal carina. These catheters were introduced through the endotracheal tube and positioned into the left and right main bronchi at the beginning of the tracheal resection. Then, conventional ventilation was replaced by high-frequency jet ventilation (HFJV) with different ventilatory variables for each lung. During two-lung jet ventilation there was good oxygenation, normocapnia and no cardiovascular complications. The principle advantage of using two separate high-frequency ventilators is that it allows for maximum ventilatory efficiency with lungs of different compliance. PMID- 8425246 TI - Hyperkalaemia: a complication of warm heart surgery. AB - A case is presented of hyperkalaemia (13.6 mEq.L-1) occurring during cardiopulmonary bypass using warm blood cardioplegia (K+ 40-60 mEq.L-1). Treatment with epinephrine, calcium chloride, sodium bicarbonate, and furosemide reduced K+ to 6.5 mEq.L-1 within 30 min and myocardial performance was enhanced with amrinone and cardiac rhythm was controlled with A-V segmental pacing. It is believed that the hyperkalaemia resulted from a combination of the surgical procedure (mitral valve replacement) and the use of warm cardioplegia. The purpose of this report is to increase the awareness of the possibility of hyperkalaemia with warm cardioplegia and to describe a successful therapeutic regimen. PMID- 8425247 TI - Consequences of misfilling contemporary vaporizers with desflurane. AB - Desflurane is a volatile anaesthetic that combines low blood gas solubility (blood/gas partition coefficient = 0.42 at 37 degrees C), moderate potency (MAC = 6-7%), and high volatility (vapour pressure = 681 mmHg at 20 degrees C, boiling point = 23.5 degrees C). The volatility and potency of desflurane prevent its safe use in vaporizers of traditional design. We present a mathematical model which demonstrates the potential for desflurane overdose if contemporary vaporizers are misfilled with desflurane. The most hazardous filling error occurs if an enflurane vaporizer is misfilled with desflurane. The calculated desflurane output of a misfilled enflurane vaporizer at a dial setting of 1% and a temperature of 22 degrees C is 57.8%, or 9.6 MAC. For misfilled enflurane, isoflurane, and halothane vaporizers at dial settings equivalent to one MAC at 22 degrees C, the calculated desflurane output is 14.0, 10.2, and 7.8 MAC, respectively. We conclude that the safe delivery of desflurane will require engineering safeguards, additional monitoring, and education of the anesthesia community. PMID- 8425248 TI - Vaporizer overfilling. PMID- 8425249 TI - Anaesthetists and the right to die. PMID- 8425250 TI - Four-centre study of anaesthetic outcomes. PMID- 8425251 TI - Cost of soda lime. PMID- 8425252 TI - An accidental subdural injection of a local anaesthetic resulting in respiratory depression. PMID- 8425253 TI - Anaesthetic management of a parturient with thrombocytopenia using thrombelastography and sonoclot analysis. PMID- 8425254 TI - Distribution and metabolism of aflatoxin B1 in the marmoset monkey (Callithrix jacchus). AB - Whole-body autoradiography of [3H]aflatoxin B1 (3H-AFB1) in marmoset monkeys (Callithrix jacchus) showed a localization of bound labelling, in addition to the liver, in the nasal olfactory and respiratory mucosa and the mucosa of the nasopharyngeal duct, the pharynx, the larynx, the trachea and the oesophagus. In vitro microautoradiography of these tissues incubated with 3H-AFB1 showed a localization of bound radioactivity in some cells in the epithelial linings of the tissues. This binding was abolished when the incubations were performed in the presence of the cytochrome P-450 inhibitor metyrapone. These results indicate a cytochrome P-450-dependent bioactivation of AFB1 in some structures in the epithelia of the nasal olfactory mucosa and the upper respiratory and alimentary pathways, in addition to the liver, in the marmoset monkey. Quantitation of the in vitro formation of tissue-bound radioactivity from the 3H-AFB1 showed that the nasal olfactory mucosa had the highest capacity to form bound metabolites among the tissues examined. Liquid chromatography of lipid-soluble metabolites formed by the nasal olfactory mucosa and the liver showed that aflatoxin M1 (AFM1) was the major metabolite. AFM1 was also the major metabolite found in the liver in vivo. In the pigmented tissues of the marmosets there was an accumulation of nonmetabolized AFB1. This can be related to a melanin-affinity of AFB1. The described tissues with a capacity to accumulate and metabolize AFB1 may be potential targets for the carcinogenicity of this substance in the marmoset monkey. PMID- 8425255 TI - Flow cytometric immunofluorescence assay for quantification of cyclobutyldithymine dimers in separate phases of the cell cycle. AB - Quantitative immunofluorescence assays for the measurement of cyclobutyldithymine dimers (T <> T) based on computer-assisted immunofluorescence microscopy have recently been described. Here we present a modified assay for T <> T based on flow cytometry. This method has the advantage that T <> T can be quantified in separate phases of the cell cycle by the fluorescent counterstaining of nuclear DNA and subsequent selection on DNA content. The H3 monoclonal antibody directed at T <> T binds to partially denatured DNA in situ. The antibody is labeled with fluorescein isothiocyanate (FITC) and DNA is stained with the intercalating dye 7 amino-actinomycin D. FITC fluorescence increases linearly with dose of UV-C radiation (up to 45 J/m2) of cultured human fibroblasts. A linear fluorescence dose relationship was also found for epidermal cells of SKH:HR1 hairless mice after in vivo irradiation with UV-B (FS40 sunlamp, up to 3750 J/m2). This technique allows a quick assessment of UV damage levels in 10,000s of cells and makes immunofluorescence of DNA damage more accessible to other research groups. PMID- 8425256 TI - Cellular damage by ferric nitrilotriacetate and ferric citrate in V79 cells: interrelationship between lipid peroxidation, DNA strand breaks and sister chromatid exchanges. AB - Ferric nitrilotriacetate (Fe-NTA) and ferric citrate (Fe-citrate) were used to study the cellular damage mediated by iron overload with respect to cytotoxicity, lipid peroxidation, DNA strand breaks and sister chromatid exchanges (SCE). At non-toxic concentrations, Fe-NTA induced lipid peroxidation, DNA strand breaks and SCE in a dose- and time-dependent manner. Comparing the time courses of the different events, the increase in lipid peroxidation seems to be associated with the generation of DNA strand breaks, since both types of cellular damage were observed after 1-3 h of incubation. In contrast, the induction of SCE was low after 24 h and increased after 48 and 72 h treatment, indicating the requirement of other types of DNA damage. Fe-citrate was inactive in the induction of lipid peroxidation and SCE, and no significant number of DNA strand breaks were generated, as determined by the alkaline unwinding method. Our results suggest that the induction of lipid peroxidation and DNA strand breaks by iron overload depend on special features of the iron complex applied, while the chromosomal and genetic effects require site specific DNA damage dependent on intracellular iron metabolism. It is concluded that iron overload is able to induce genetic damage in intact cells provided that iron is present in a bioavailable form. PMID- 8425257 TI - Genotoxicity characteristics of reverse diol-epoxides of chrysene. AB - Trans-3,4-dihydroxy-3,4-dihydrochrysene (chrysene-3,4-diol), a major metabolite of chrysene, is further metabolized by rat liver enzymes to products which effectively revert the his- Salmonella typhimurium strain TA98 to histidine prototrophy, but are only weakly mutagenic in strain TA100 and in Chinese hamster V79 cells (acquisition of resistance to 6-thioguanine). The liver enzyme mediated mutagenicity of chrysene-3,4-diol is substantially enhanced in the presence of 1,1,1-trichloropropene 2,3-oxide, an inhibitor of microsomal epoxide hydrolase. The predominant metabolites of chrysene-3,4-diol, namely the anti- and syn isomers of its 1,2-oxide (termed reverse diol-epoxides), proved to be extraordinarily effective mutagens in S.typhimurium strain TA98, but were only moderately active in strains TA100 and TA104, and in the SOS induction in Escherichia coli PQ37. These genotoxicity spectra in bacteria are completely different from those observed with the bay-region diol-epoxides of chrysene and 3 hydroxychrysene. In V79 cells, the reverse diol-epoxides formed low levels of DNA adducts and were very weak inducers of gene mutations. In M2 mouse prostate cells, however, high numbers of transformed foci were induced by chrysene-3,4 diol and its diastereomeric 1,2-oxides. Chrysene-3,4-diol was somewhat more potent than chrysene-1,2-diol. The potency of both reverse diol-epoxides was similar to that of the syn-diastereomers of the bay-region diol-epoxides of chrysene and 3-hydroxychrysene, but lower than that of their anti-diastereomers. The reverse diol-epoxides of chrysene, unlike the bay-region diol-epoxides, were inactivated by purified microsomal epoxide hydrolase. Noteworthy findings were also made with regard to the chemical stability of the diol-epoxides in buffer, determined from the decline in mutagenicity after preincubation in the absence of the target cells. Despite its lower delta Edeloc/beta value for the formation of the benzylic carbocation, anti-chrysene-3,4-diol 1,2-oxide was shorter-lived (t1/2 = 46 min) than anti-chrysene-1,2-diol 3,4-oxide (t1/2 = 74 min). Unlike other investigated diastereomeric pairs of diol-epoxides, it was also shorter lived than its syn-diastereomer (t1/2 = 340 min). PMID- 8425258 TI - Development of monoclonal antibodies specific for 1,N2-ethenodeoxyguanosine and N2,3-ethenodeoxyguanosine and their use for quantitation of adducts in G12 cells exposed to chloroacetaldehyde. AB - Monoclonal antibodies specific for N2,3-ethenodeoxyguanosine (N2,3-epsilon dGuo) and 1,N2-ethenodeoxyguanosine (1,N2-epsilon dGuo) were developed. In a competitive ELISA, 50% inhibition of binding of the N2,3-epsilon dGuo specific antibody (ETH1) was achieved with 18 fmol of N2,3-epsilon dGuo. Fifty per cent inhibition of the 1,N2-epsilon dGuo-specific antibody (ETH2) required 11 pmol 1,N2-epsilon dGuo. Immunoassays for N2,3-epsilon dGuo and 1,N2-epsilon dGuo in single-stranded DNA were developed using these antibodies. The immunoassays could detect as little as 48 fmol of N2,3-epsilon dGuo or 340 fmol 1,N2-epsilon dGUO in 25 micrograms of single stranded DNA. These assays and previously developed immunoassays for 1,N6-ethenodeoxy-adenosine (1,N6-epsilon dAdo) and 3,N4 ethenodeoxycytidine (3,N4-epsilon dCyd) were used to measure etheno adduct levels in DNA of cells exposed to chloroacetaldehyde. The cells used were V79 cells with an inactivated hprt gene and a single copy of the bacterial gpt gene (G12 cells). The most abundant etheno adduct was 1,N6-epsilon dAdo, followed by 3,N4-epsilon dCyd and N2,3-epsilon dGuo. 1,N2-epsilon dGuo was not detected in chloro acetaldehyde-treated G12 cells. Chloroacetaldehyde was also shown to be mutagenic in these same cells. PMID- 8425259 TI - Expression of cytochrome P450 in LEC rats during the development of hereditary hepatitis and hepatoma. AB - The expression of 14 forms of cytochrome P450 in the liver as well as changes in the testosterone hydroxylation activities of hepatic microsomes were investigated during the development of hepatitis in Long-Evans Cinnamon (LEC) rats. P4501A1 and -1A2 (3-methylcholanthrene-inducible forms) and P4502B1 and -2B2 (phenobarbital-inducible forms) were barely detected in the hepatic microsomes of male and female LEC rats. In immature male rats, the levels of male-specific forms (P4502C11 and -2C13) were higher in LEC rats than in control Long-Evans Agouti (LEA) rats. P4502C11 appeared in female LEC rats from 4 to 16 weeks of age, reflecting that testosterone 2 alpha- and 16 alpha-hydroxylation activities were detected at significant levels in female LEC rats. In immature female rats, the level of P4502C12 (a major female-specific form) was higher in LEC rats than in LEA rats. The level of P4502C13 in male LEC rats and that of P4502C12 in female LEC rats decreased markedly with ageing or during the development of hepatitis. The level of P4503A2 (a male-predominant form) was especially high in immature male and female LEC rats, reflecting that both rats had high 2 beta- and 6 beta-hydroxylation activities toward testosterone. These sex-specific forms are regulated by androgens and by pituitary growth hormone. Thus, there may be abnormalities of the hypothalamo-pituitary-gonadal axis in LEC rats. Furthermore, P4503A2 efficiently activates aflatoxin B1, a potent hepatocarcinogen, and the increased levels of this form in LEC rats may be related to the onset of hepatitis or liver cancer. PMID- 8425260 TI - Structure of an antimutagen from Carmona retusa leaves. AB - An antimutagenic compound was isolated from the leaves of Carmona retusa (Vahl) Masam. Its structure has been elucidated by spectral analysis to be 4-hydroxy 7,8,11,12,15,7',8',11',12',15'-decahydro-beta, psi-carotene. The results of the Micronucleus Test, an in vivo method, showed that the isolated antimutagenic compound reduced by approximately 68.4% the number of micronucleated polychromatic erythrocytes induced by tetracycline, a known mutagen. PMID- 8425261 TI - 32P-postlabelling analysis of the covalent binding of benzo[ghi]perylene to DNA in vivo and in vitro. AB - Benzo[ghi]perylene (B[ghi]P) is a polycyclic aromatic hydrocarbon (PAH), present in complex combustion products, and evidence for its carcinogenic activity in experimental animals is equivocal, and yet it has demonstrable mutagenic activity in vitro. In order to investigate the possible DNA binding properties of B[ghi]P, groups of male Parkes mice were treated topically with 1.0 mumol of B[ghi]P. Mice were killed up to 3 months after treatment, DNA was isolated from the treated areas of skin and analysed for adducts by 32P-postlabelling. Maximum levels of binding (0.57 fmol/microgram DNA) were detected 2 days after treatment and adducts were found to persist for at least 12 weeks after treatment, a property of many PAHs with known tumor-initiating activity. B[ghi]P also became bound to DNA in vitro in the presence of 3-methylcholanthrene-induced rat liver microsomal preparations. When chromatographed on PEI-cellulose, the major adducts formed by B[ghi]P in vivo and in vitro appeared to be identical. However, they were found to be different when compared by reversed-phase HPLC. These differences might explain, in part, the differences in the biological activity of B[ghi]P in vivo and in vitro. The behaviour of B[ghi]P when present in a mixture was also examined. B[ghi]P was applied topically to mouse skin with six other PAHs at a dose level of 0.25 mumol/PAH/mouse. DNA isolated 24 h after treatment was analysed for adducts by 32P-postlabelling. Whilst the total level of binding was 30% lower than expected from the sum of the binding levels that resulted when the hydrocarbons were applied singly, the formation of B[ghi]P-DNA adducts did not appear to be inhibited. The results have demonstrated that B[ghi]P has significant DNA binding ability in vivo and in vitro and on the basis of its DNA binding ability in mouse skin it would be predicted to be at least a weak tumour initiator. The formation of DNA adducts by B[ghi]P when present in an artificial mixture of PAHs suggest that B[ghi]P may contribute to the DNA binding activity of more complex carcinogenic mixtures. PMID- 8425262 TI - Covalent binding of polycyclic aromatic hydrocarbon components of coal tar to DNA in mouse skin. AB - Treatment of mouse skin with coal tar is known to initiate tumour formation, with the carcinogenic activity associated mainly with polycyclic aromatic hydrocarbons (PAHs). A sample of pharmaceutical coal tar was analysed by gas chromatography and 19 major PAHs were identified. 32P-postlabelling analysis was used to characterize those PAHs that are responsible for the DNA binding of coal tar and, by inference, its biological activity. PAHs were grouped according to their reported carcinogenic activities and applied as mixtures to mouse skin. Group A contained all of the 19 PAHs, group B seven PAHs for which there is sufficient evidence for carcinogenicity and group C 12 PAHs with only limited or inadequate evidence of carcinogenicity in experimental animals. 32P-Labelled DNA adducts formed by coal tar were resolved on TLC into a pattern of three discrete spots (2, 4 and 6) and four areas of diffuse radioactivity (1, 3, 5 and 7). By comparison of the pattern of adducts formed by coal tar with those formed by the synthetic mixtures it appeared that PAHs in group B formed coal tar-DNA adduct spots 4 and 6, and that adduct spot 2 was formed by PAHs in group C. Attempts to identify those PAHs responsible for the formation of coal tar-DNA adducts 4 and 6 were made by comparing the chromatographic mobilities of 32P-labelled coal tar derived DNA adducts formed in mouse skin, using TLC and HPLC, with those formed by PAHs in group B. As benzo[ghi]perylene (B[ghi]P), a component of group C, has been demonstrated to exhibit significant DNA binding ability previously, the chromatographic mobility of coal tar-DNA adduct spot 2 was compared to that of the major DNA adducts formed by B[ghi]P in vivo and in vitro. It appeared that coal tar adduct spot 2 was the major adduct formed by B[ghi]P in vitro and that benzo[a]pyrene, benzo[b]fluoranthene, benzo [j]fluoranthene and benzo[k]fluoranthene contributed to the formation of adduct spot 6. None of the PAHs examined appeared to be responsible for the formation of adduct spot 4. PMID- 8425263 TI - Analysis of activated protooncogenes in B6C3F1 mouse liver tumors induced by ciprofibrate, a potent peroxisome proliferator. AB - Liver tumors from B6C3F1 mice induced by the potent peroxisome proliferator ciprofibrate, a hypolipidemic drug, were evaluated for the presence of transforming genes by the nude mouse tumorigenicity assay. As reported earlier, the tumors were not activated by a point mutation in codon 61 of H-ras. Two of the eight tumors examined contained a mutation in codon 13 or an H-ras gene mutated in codon 117. Screening of another 23 ciprofibrate-induced liver tumors by oligonucleotide hybridization analysis and direct DNA sequencing resulted in the identification of three tumor DNA samples with point mutations in codon 117 of the H-ras gene. In addition, another tumor sample contained a K-ras gene with a mutation in codon 61. Mutations in these codons have been seen only rarely in chemically induced liver tumors from this mouse strain. Of 15 spontaneous B6C3F1 liver tumors screened in the same manner, one exhibited a K-ras gene activated by a mutation in codon 13 and a second contained an H-ras gene activated by a mutation in codon 117. These ras gene mutations have not been reported previously from spontaneous liver tumors. The frequency and spectrum of ras oncogene mutations characterized in ciprofibrate-induced liver tumors differ significantly from the frequency and pattern identified in spontaneously occurring liver tumors. The results of this study with a limited number of samples suggest that ras protooncogene activation or activation of other protooncogenes that can be detected by the nude mouse tumorigenicity assay are not frequent events in the mechanism of carcinogenicity of the peroxisome proliferator ciprofibrate. However, the lower frequency and distinct pattern of H-ras mutations observed in these tumors disprove the assumption of promotion of spontaneous hepatocarcinogenesis by ciprofibrate. PMID- 8425264 TI - Smoking-related DNA adducts: 32P-postlabeling analysis of 7-methylguanine in human bronchial and lymphocyte DNA. AB - 7-methylguanine DNA adducts were determined in macroscopically normal bronchial specimens and peripheral blood lymphocytes of 20 patients undergoing pulmonary surgery. A recently developed 32P-postlabeling assay was applied with anion exchange chromatography as an adduct enrichment method. The material consisted of 13 smokers and 7 non-smokers. The mean bronchial 7-methylguanine levels of 11 smokers and 6 non-smokers were 17.3 and 4.7 adducts/10(7) nucleotides. In lymphocyte DNA, the respective mean levels were 11.5 and 2.3 adducts/10(7) nucleotides. The bronchial DNA adduct levels in smokers were statistically higher than those in non-smokers. Among 5 smokers, for whom both bronchial and lymphocyte DNA was available, 7-methylguanine levels correlated in the two tissues (r = 0.77). PMID- 8425265 TI - Use of fluorescently tagged DNA and an automated DNA sequencer for the comparison of the sequence selectivity of SN1 and SN2 alkylating agents. AB - This paper describes the application of the novel nonradioactive technique for studying the sequence selectivity of selected alkylating agents. N-Nitroso-N methylurea (MNU) and N-methyl-N'-nitro-nitrosoguanidine (MNNG) were chosen from the SN1 group of alkylating agents. Dimethyl sulphate (DMS) was used to represent alkylation profile produced by the SN2 compounds. Results of SN1 compounds indicated that in a run (G)3 the latter two Gs are more susceptible to alkylation than the most 5' G. Moreover, in a GG sequence the 3' G seems to be more alkylated. This effect is more evident when the GG site was preceded by a 5' pyrimidine. These findings suggest that a regio-selective mechanism, rather than the formation of diazonium ions, accounts for DNA alkylation by SN1 compounds. On the other hand, DMS showed preferential alkylation of the 5' end in a (G)3 run. However, at GG sequences no clear preferred site of alkylation could be distinguished. Lack of specificity of SN2 compound would seem to suggest that other factors as well as the primary DNA structure may play a role in determining the extent of alkylation at a certain site. PMID- 8425266 TI - Different inhibition of DNA synthesis by transforming growth factor beta and phenobarbital on GST-P-positive and GST-P-negative hepatocytes. AB - Hepatocyte resistance against inhibition of DNA synthesis by transforming growth factor beta 1 (TGF-beta 1) was studied in vitro. Hepatocytes were isolated from rats that had received diethylnitrosamine (DEN) for 6 weeks. The effect of TGF beta 1 and phenobarbital (PB) on DNA synthesis in different cell populations was studied using bromodeoxyuridine (BrdU) incorporation and placental glutathione S transferase (GST-P) as markers. It was found that GST-P-positive cells were resistant to the growth inhibitory effect of TGF-beta 1 and PB, whereas GST-P negative cells were inhibited. It is concluded that resistance to TGF-beta 1 dependent growth control may develop early during DEN-induced hepatocarcinogenesis. PMID- 8425267 TI - Activation of ribosomal protein S2 gene expression in a hamster model of chemically induced oral carcinogenesis. AB - 7,12-Dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis of the hamster buccal pouch has been an excellent model for the study of squamous cell carcinogenesis in human head and neck cancer. Using a differential hybridization of cDNA cloning technique, we isolated a cDNA clone that is expressed in N-ras transformed PA-1 cells but poorly expressed in non-tumorigenic PA-1 cells; the cDNA codes for the human ribosomal S2 gene product. To define the involvement of S2 gene expression during carcinogenesis in this animal model, we used in situ hybridization technique with non-radioactive digoxigenin-11-dUTP-labeled cDNA. S2 gene was expressed at low levels in basal and suprabasal cell layers of the epidermis in the control, but showed marked elevation throughout the epidermis other than the keratin layer in samples treated for 4 or 8 weeks; S2 was highly expressed in all malignant squamous cell carcinoma cells resulting from DMBA treatment for 16 weeks. As tumors progress from normal epithelium to squamous cell carcinomas, mRNA of the S2 gene was not only elevated sequentially, but also demonstrated the marked heterogeneity among transformed populations, particularly in dysplastic lesions and squamous cell carcinomas. The S2 gene was expressed in a stage-specific manner in the hamster tumor model; S2 could be useful as a neoplastic marker for the detection of certain epithelial origin of tumors and premalignant lesions as well. PMID- 8425268 TI - Immortalization phenotype dissociated from the preneoplastic phenotype in mouse mammary epithelial outgrowths in vivo. AB - Mouse mammary epithelial cells (MMEC) isolated from normal virgin BALB/c female mice and grown in cell culture for various lengths of time were injected into the mammary fat pads of syngenic mice. Of the ductal outgrowths which resulted from the injected MMEC, four gave rise to outgrowths that were serially transplanted beyond the lifetime of normal ductal outgrowths. The lifetime of normal ducts is five or six transplant generations. The four ductal outgrowth lines, termed EL for 'extended life', have been serially transplanted for 7, 9, 13 and 14 transplant generations. The outgrowths are predominantly ductal in morphology, do not exhibit intraductal epitheliosis characteristic of ductal hyperplasias, are ovarian dependent for growth and are responsive to prolactin-mediated alveolar differentiation. Three of the EL lines, EL5, 7 and 11 have not produced any tumors spontaneously (0/64) and only one tumor after dimethylbenz[a]anthracene (DMBA) treatment (1/30). The fourth line, EL12, differs from the other three in the presence of a limited degree of alveolar differentiation. The EL12 line has not produced any spontaneous tumors (0/23) but is somewhat more responsive to DMBA (3/10). We interpret the EL lines (at least EL11 and EL12) to represent cell populations where the immortalized phenotype is dissociated from the hyperplastic phenotype which is characteristic of mouse mammary preneoplastic populations. The tumor suppressor gene, p53, is not overexpressed in the EL ductal outgrowths. To our knowledge, this is the first example of cell populations in vivo that are immortalized but otherwise normal. As such, they may represent the earliest stage observable in the genesis of mouse mammary tumors and provide unique cell populations to examine molecular alterations associated with the property of immortality. PMID- 8425269 TI - Enhancing effect of ethanol on esophageal tumor development in rats by initiation of diethylnitrosamine. AB - The effects of ethanol on initiation of esophageal tumorigenesis by diethylnitrosamine (DEN) were investigated in male F344 rats. Animals were administered 50 p.p.m. DEN plus 10% ethanol (Group 1), 33 p.p.m. DEN (Group 2), 50 p.p.m. DEN (Group 3) or 10% ethanol (Group 4) in the drinking water for the first 8 weeks, and were then maintained on basal diet and tap water for up to 104 weeks. The concentration of 33 p.p.m. DEN in Group 2 was an adjustment to the anticipated intake in Group 1. Consequent total intakes of DEN in Groups 2 and 3 were respectively 80% and 134% of that in Group 1. Histopathological examination of esophagus tissue after final sacrifice revealed incidences of papillomas and/or carcinomas in Groups 1, 2 and 3 to be 57.7, 3.8 and 10.7% respectively. No esophageal tumors occurred in Group 4. The incidence of hyperplasia was 100% in Group 1 in contrast to 46.2% in Group 2, 57.1% in Group 3 and 3.6% in Group 4. Thus, the incidences of esophageal proliferative lesions were significantly higher in Group 1 than in Groups 2 or 3. Our results clearly indicate that ethanol has an enhancing effect on the development of esophageal tumors induced by DEN in rats, when administered during the initiation phase. PMID- 8425270 TI - Radical oxidation reactions of the purine moiety of 2'-deoxyribonucleosides and DNA by iron-containing minerals. AB - The radical oxidation capability of several classes of iron minerals, including biotite, hematite, magnetite, minette, nemalite, pyrite, vivianite and two chrysotiles (asbestos), was investigated by using a double experimental approach. One involved the electron spin resonance spin-trapping measurement of organic radicals obtained by the reaction of activated oxygen species, released upon incubation of the minerals in phosphate buffered solutions with formate used as the target molecule. In addition, the formation of mineral-mediated oxidation purine decomposition products, including 7,8-dihydro-8-oxo-2'-deoxyguanosine and 7,8-dihydro-8-oxo-2'-deoxyadenosine, was searched within nucleosides and DNA by using specific and sensitive HPLC electrochemical assays. Emphasis was placed on the mechanistic aspects of the radical oxidation reactions involved in the formation of the two C(8) hydroxylated purine decomposition products. PMID- 8425271 TI - DNA adduct formation after oral administration of 2-nitrofluorene and N-acetyl-2 aminofluorene, analyzed by 32P-TLC and 32P-HPLC. AB - DNA adducts have been detected in laboratory animals after exposure to carcinogens as well as in human populations with known or suspected risk of developing cancer. Examples are smokers, coke and aluminium workers, urban citizens and roofers. The formation of DNA adducts is an early event in carcinogenesis which can be used for measuring target dose and as a biomarker for genotoxic risk. A method of analyzing 32P-postlabelled DNA adducts on reverse HPLC with on-line detection of 32P has been developed. The method permits direct injection of the 32P-postlabeling mixture into the analytical system without prior purification with background radioactivity on a low level. The method can be used in parallel with TLC analyses of 32P-postlabelled DNA adducts to improve the analytical capacity. The time for analysis of a typical single sample by HPLC and TLC is 30-60 min and 6-24 h respectively. A high (2 M) salt concentration in the HPLC eluent reduces the 32P background considerably. Also the peak tailing was substantially diminished, giving an ability to separate DNA adducts equal to or better than the TLC method. The method has been applied to 2-nitrofluorene (NF), a carcinogenic air pollutant, and N-acetyl-2-aminofluorene (AAF), a model carcinogen which is also a metabolite of NF. A number of DNA adducts are formed in the livers of rats. After oral administration of AAF and NF, DNA adducts in the liver have been characterized as dG-C8-AF and dG-C8-AAF. The major DNA adduct found in both NF- and AAF-administered animals was dG-C8-AF. The described HPLC method can, with minor adjustments, generally be used to analyze 32P-postlabelled DNA adducts. PMID- 8425272 TI - Mutagenic activation of IQ, PhIP and MeIQx by hepatic microsomes from rat, monkey and man: low mutagenic activation of MeIQx in cynomolgus monkeys in vitro reflects low DNA adduct levels in vivo. AB - Cooked meat, poultry and fish contain a number of mutagenic and carcinogenic heterocyclic amines, including 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2 amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP). In the present study we examined the capacity of hepatic microsomes from Fischer 344 rats, cynomolgus monkeys and humans to metabolically activate IQ, MeIQx and PhIP in vitro using the Ames Salmonella mutagenicity assay. The mutagenic activation of IQ was similar among the three species; however, there were significant differences among the species in the activation of PhIP and MeIQx. Liver microsomes from humans showed the greatest capacity to activate PhIP and MeIQx, followed by rats, and then monkeys. The largest differences between the species were observed when MeIQx was used as the mutagen. MeIQx-DNA adducts formed in vivo were then compared among rats and monkeys given MeIQx by gavage (20 mg/kg/day, 10 doses). 32P-Postlabeling analysis, carried out under intensification conditions, was used to examine MeIQx DNA adducts in the liver, kidney, heart, colon and white blood cells. MeIQx-DNA adducts were highest in all tissues examined from male rats, followed by female rats, and much lower in monkeys. In the liver, the total MeIQx-DNA adduct levels of monkeys were approximately 19 and approximately 10 times lower than in male and female rats respectively. In extrahepatic tissues, the differences in MeIQx DNA adduct levels between monkeys and rats were even greater. The results suggest that the low level of MeIQx-DNA adducts found in vivo in cynomolgus monkeys reflects a low capacity to activate MeIQx via the hepatic cytochrome P450 monooxygenase system. PMID- 8425273 TI - Effects of five phorbol esters on gap junctional intercellular communication, morphological transformation and epidermal growth factor binding in Syrian hamster embryo cells. AB - The effects of 12-O-tetradecanoylphorbol-13-acetate (TPA), 12-deoxyphorbol-13 phenylacetate (DOPP), 12-deoxyphorbol-13-phenylacetate-20-acetate (DOPP A), sapintoxin D (SAP D) and sapintoxin A (SAP A) on the decrease in [125I]epidermal growth factor (EGF) binding (indicating protein kinase C activation), suppression of gap junctional intercellular communication (GJIC) and induction of morphological cell transformation (MCT) in Syrian hamster embryo (SHE) cells were investigated. All five phorbol esters were found to reduce [125I]EGF binding in early passage SHE cells at comparable concentrations. DOPP A was approximately 10 fold less potent in decreasing GJIC compared to the other phorbol esters in early passage SHE cells, while the compounds showed less difference in suppressing GJIC in the phorbol ester sensitive SHE cell line BPNi. The decreases in [125I]EGF binding and GJIC were found to be transient in the continuous presence of phorbol esters. All phorbol esters induced MCT in early passage SHE cells, but DOPP and DOPP A were approximately 10-fold less potent than TPA, SAP D and SAP A. Thus, there seems to be some degree of correlation, but not to a full extent, between the ability of the phorbol esters to activate PKC, decrease GJIC and to induce MCT. The results do not suggest a simple relationship between PKC activation, inhibition of GJIC and the reported tumor-promoting activities of the compounds. PMID- 8425274 TI - Effects of edible oils and fatty acids on the formation of mutagenic heterocyclic amines in a model system. AB - The effects of glycerol, fatty acids and oils on the yield and species of mutagenic heterocyclic amines were studied in a model system. The addition of lipids to the model system did not affect the species of food mutagens formed, but did affect the yield of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx). When heating creatinine, glycine and glucose dissolved in water, at 180 degrees C for 10 and 30 min, 9 and 18 nmol MeIQx/mmol creatinine were formed respectively. Corresponding figures of MeIQx formed, after addition of various fatty acids or edible oils to the model system, were as follows: oleic acid (9 and 11 nmol MeIQx/mmol creatinine), stearic acid (16 and 19 nmol), linoleic acid (8 and 16 nmol), linolenic acid (5 and 20 nmol), corn oil (10 and 33 nmol) and olive oil (10 and 28 nmol) after heating at 180 degrees C for 10 and 30 min respectively. Addition of corn or olive oil in a model system heated at 180 degrees C for 30 min, almost doubled the yield of MeIQx formed, compared with the amount formed in a model system without fat. This increase was not observed if glycerol or a fatty acid was added to the model system. PMID- 8425275 TI - Effects of oxidative stress induced by redox-enzyme modulation on the progression stage of rat hepatocarcinogenesis. AB - Effects of oxidative stress induced by redox-enzyme modulation on the progression stage of hepatocarcinogenesis were examined by monitoring both hepatocyte injury and hepatocellular carcinoma development in F344 rats bearing preneoplastic liver nodules induced by the Cayama-Farber procedure. Redox-enzyme modulation, which included increased cytochrome P450 reductase activity induced by phenobarbital-Na (100 mg/kg, i.p. for 3 days), inhibition of DT-diaphorase by dicumarol (25 mg/kg, i.p.), depletion of glutathione by phorone (200 mg/kg, i.p.), supplementation with the Fe(III) sodium salt of EDTA (50 mg/kg, i.p.) and redox-cycling activation by menadione (50 mg/kg, i.g.), exerted no prominent hepatocyte injury within nodules but did cause slight injury in the surrounding hepatocytes in nodule-bearing rats. The same treatments induced severe hepatocyte injury in non treated normal rats. Redox-enzyme modulation performed every other week for 33 weeks significantly reduced the number of hepatocellular carcinomas developing in nodule-bearing rats. These results indicate that preneoplastic nodules are resistant to the oxidative stress induction caused by redox-enzyme modulation treatment and that, despite toxic effects in surrounding hepatocytes, no progression pressure is exerted. Indeed, the treatment rather demonstrates an inhibitory effect of the evolution of the nodules into hepatocellular carcinomas. PMID- 8425276 TI - Parents' evaluations of pre-school services for children with Down syndrome in two Scottish regions. AB - Parental evaluations of current pre-school provision for children with Down's syndrome were surveyed in two Scottish regions. Two methods were used: postal questionnaires and telephone interviews. Services covered were those provided by general practitioners, hospital specialists, health visitors, speech therapists, occupational therapists, physiotherapists, educational psychologists, home teachers, social workers and voluntary organizations. Regional differences were found in provision and in parental satisfaction with currently available services, with some of these differences being dependent on child age. Overall, parents felt they were being given insufficient professional support, with contradictory advice not uncommon. Findings indicate that if limited resources are to be used to the maximum benefit of family and child, both subjective and objective measures of the relative values of different kinds of support at different ages are urgently needed. PMID- 8425277 TI - Pre-school visually impaired children: visual stimulation and micro-computers. AB - A justification is offered for using micro-computers and specially-designed programs with severely visually impaired pre-school children. It is argued that this technology optimizes the visual environment for the child and provides immediate feedback about the correctness of his responses to the stimuli displayed on the screen. Some of the problems encountered in such use are described, with examples taken from sessions in which teachers are working with children as young as 2 years of age, using teaching/learning sequences designed to promote and exercise visual perception skills. Among the issues addressed are the nature of the language interactions between child and adult, the effects of altering the complexity of the tasks, and the possible value of formal task analysis as a means of enabling the teacher to pinpoint critical stages in the learning process. PMID- 8425278 TI - BCG vaccination scars: incidence and acceptance amongst British high-school children. AB - In order to determine the incidence and acceptance of BCG scars, 287 high-school children of different ethnic origins, in a London district, were examined for their BCG scars and interviewed for self-appraisal of their scars 6-30 months after vaccination. BCG scars developed in a high proportion of children (89.5%). There was a female predominance among the 10.5% of children who did not develop scars (girls 12.8%, boys 5.9%, P < 0.05). Hypertrophic scars (defined as the largest diameter of scar > 13.24 mm, [i.e. 2 SD above mean]) were found in 3.11% and hypopigmented scars in 67.8% of the children and both tended to occur amongst hyperpigmented races. A high proportion of children found the scars unacceptable (23.4%), mostly girls (girls 35%, boys 7.8%, P < 0.0004) and they showed a preference for other sites including inner aspect of arm and buttock for vaccination. PMID- 8425279 TI - Re: The school entry medical examination--what teachers think of it? PMID- 8425280 TI - Long-term benefit of 1-year amiodarone treatment for persistent complex ventricular arrhythmias after myocardial infarction. AB - BACKGROUND: In the Basel Antiarrhythmic Study of Infarct Survival trial, low-dose amiodarone improved 1-year survival in patients with asymptomatic complex ventricular arrhythmias persisting 2 weeks after myocardial infarction. To assess whether this beneficial effect persisted despite discontinuation of amiodarone after 1 year, the long-term outcomes of all patients of the amiodarone-treated group (initially n = 98) and those of the control group (n = 114) were assessed. METHODS AND RESULTS: After a mean follow-up of 72 (55-125) months, information on 96% of patients (203 of 212) was obtained regarding survival or cause of death. The probability of death after 84 months according to actuarial life-table analysis (Kaplan-Meier) was 30% for the amiodarone-treated patients and 45% for control patients. For the total follow-up, mortality remained significantly lower in the amiodarone group versus the control group regarding all deaths (p = 0.03) as well as cardiac death (p = 0.047). This mortality reduction was entirely due to the first-year amiodarone effect, since there was no significant mortality difference between groups when considering survival after discontinuation of amiodarone only. CONCLUSIONS: These data suggest that the beneficial effect of amiodarone on survival in this high-risk group of patients persists for several years. In addition, the results stress the importance of early treatment after myocardial infarction, whereas the rate of sudden death and all cardiac death is low (1.6% and 4.1% per year, respectively) during late follow-up and therefore may not warrant further therapy. PMID- 8425281 TI - Determinants of predicted efficacy of antiarrhythmic drugs in the electrophysiologic study versus electrocardiographic monitoring trial. The ESVEM Investigators. AB - BACKGROUND: The Electrophysiologic Study Versus Electrocardiographic Monitoring (ESVEM) study was designed to compare the accuracy of predictions of antiarrhythmic drug efficacy made by electrophysiological study (EPS) with those made by Holter monitoring (HM) combined with exercise testing. The present study describes the baseline characteristics and the response to drug efficacy tests of 486 randomized subjects. METHODS AND RESULTS: Patients with ventricular tachyarrhythmias were randomly assigned to undergo serial testing of up to six antiarrhythmic drugs by either EPS (EPS limb) or HM and exercise testing (HM limb). Efficacy predictions were achieved in 108 of 242 patients in the EPS limb (45%) and in 188 of 244 patients (77%) in the HM limb. Left ventricular ejection fraction (LVEF) < 0.25 and presence of coronary artery disease were negative correlates (p < 0.10) of drug efficacy predictions in the EPS limb. In the HM limb, LVEF was the lone univariate correlate of efficacy, although it was only marginally significant (p = 0.107). A multivariate model selected assessment by HM and higher LVEF as independent predictors (p < 0.05) of drug efficacy. The drug evaluation process required an actuarial median time of 25 days in the EPS limb and 10 days in the HM limb (p < 0.0001). CONCLUSIONS: 1) Drug efficacy predictions are achieved more frequently by HM than by EPS. 2) Assessment by HM and severity of left ventricular dysfunction are independent correlates for a drug efficacy prediction. 3) The duration of drug testing is considerably shorter for the HM method. PMID- 8425282 TI - Residual coronary reserve despite decreased resting blood flow in patients with critical coronary lesions. A study by technetium-99m human albumin microsphere myocardial scintigraphy. AB - BACKGROUND: Experimental data demonstrate the persistence of a transmural vasodilator reserve in the face of depressed resting myocardial perfusion. The present study was designed to determine whether resting myocardial hypoperfusion indicates exhausted coronary reserve (CR). METHODS AND RESULTS: Fifteen patients with stable angina, isolated left anterior descending coronary artery (LAD) stenosis, and no previous myocardial infarction were evaluated by means of 99mTc human albumin microsphere scintigraphy. Regional myocardial perfusion and CR were assessed at baseline and after LAD papaverine (10-12 mg) by means of two microsphere injections in the left ventricle and compared with five normal subjects. Two 300-second scans were obtained with a mobile gamma camera positioned in the 70 degrees left anterior oblique projection; actual microsphere distribution after papaverine was obtained by image subtraction. The two arterial input functions (basal and papaverine) were measured from the first-pass time activity curves and validated with the reference arterial sample technique. From the comparison of circumferential profile analysis between patients and normal subjects, nine patients (group 1A) showed perfusion defects at rest (reduction of percent radioactivity below 2 SD of normal subjects) in the LAD territory, and the other six (group 1B) showed homogeneous perfusion. CR (papaverine/resting perfusion) was 3.8 +/- 0.2 and 1.51 +/- 0.27 in normal subjects and in ischemic patients, respectively (p < 0.01). Despite resting hypoperfusion, group 1A showed a papaverine-recruitable CR similar to that of group 1B (1.57 +/- 0.33 and 1.43 +/- 0.16, respectively, p = NS). CONCLUSIONS: In patients with stable angina pectoris, isolated LAD stenosis, and no previous myocardial infarction, microsphere scintigraphy disclosed a high incidence of resting perfusion defects; in those patients, a residual CR was observed despite decreased resting blood flow. PMID- 8425283 TI - Selection of the optimal nonexercise stress for the evaluation of ischemic regional myocardial dysfunction and malperfusion. Comparison of dobutamine and adenosine using echocardiography and 99mTc-MIBI single photon emission computed tomography. AB - BACKGROUND: The mechanisms of action of exercise-simulating and vasodilator stressors support their combination with imaging techniques that evaluate left ventricular function and perfusion, respectively. However, reported accuracies of either pharmacological stress together with two-dimensional echocardiography (2DE) or single photon emission computed tomography (SPECT) of myocardial perfusion are similar. The purpose of this study was to establish the optimal stress for each imaging technique by comparing the results of digitized 2DE and 99mTc-methoxyisobutyl isonitrile (MIBI) SPECT using both dobutamine and adenosine stresses in the same patients and conditions. METHODS AND RESULTS: Ninety-seven consecutive patients without evidence of previous infarction undergoing coronary angiography for clinical indications were studied prospectively. Dobutamine was infused during clinical, ECG, and echocardiographic monitoring in dose increments from 5 to 40 micrograms.kg-1.min-1. Adenosine was infused under the same conditions in doses of 0.10, 0.14, and 0.18 mg.kg-1.min-1. For each protocol, the end points were achievement of peak dose, development of severe ischemia, or intolerable side effects. At peak stress, 20 mCi of MIBI was injected, and SPECT imaging was performed 2 hours later; abnormal poststress images were compared with resting SPECT: Digitized 2DE images were compared qualitatively before, during, and after stress in a cine-loop display. Significant coronary disease (n = 59 patients) was defined by the quantification of > 50% stenosis in a major epicardial vessel. The sensitivity of adenosine 2DE was 58%, less than those of adenosine MIBI (86%, p = 0.001), dobutamine 2DE (85%, p = 0.001), and dobutamine MIBI (80%, p = 0.01). Their respective specificities were 87%, 71%, 82%, and 74% (p = NS). The accuracy of adenosine 2DE was 69%, compared with 80% for adenosine MIBI (p < 0.001), 84% for dobutamine 2DE (p = 0.001), and 77% for dobutamine MIBI (p = 0.005); the latter three did not differ significantly in either sensitivity or accuracy. CONCLUSIONS: This prospective, direct comparison of alternative pharmacological stresses in patients without myocardial infarction shows vasodilator stress scintigraphy and dobutamine stress echocardiography and scintigraphy to share equivalent levels of sensitivity. All three are significantly more sensitive than adenosine stress echocardiography. Dobutamine stress may be used for wall motion or perfusion imaging, but adenosine stress is best combined with perfusion scintigraphy. PMID- 8425284 TI - Time course of functional improvement in stunned myocardium in risk area in patients with reperfused anterior infarction. AB - BACKGROUND: The beneficial effect of coronary reflow on myocardial salvage may be assessed more accurately than in previous studies if the size of risk area is taken into account, particularly because the size of risk area varies significantly among patients. In this study, the risk area was determined with myocardial contrast echocardiography to investigate the time course of functional recovery of postischemic myocardium within the risk area in patients with reperfused anterior myocardial infarction. METHODS AND RESULTS: The study population consisted of 21 patients with anterior myocardial infarction who achieved coronary reflow within 6 hours of onset by means of thrombolysis or coronary angioplasty. Myocardial contrast echocardiography was performed with the injection of hand-agitated Haemaccel (5 ml) into the right and left coronary arteries before coronary reflow, and the risk area was defined as the area of contrast perfusion defect in the apical long-axis view. The ratio of the endocardial length of abnormal contraction (dyskinesis/akinesis) segment to that of contrast defect segment (AS/CD) was determined at days 1, 2, 3, 7, 14, and 28 of reflow. Before reflow, the length of contrast defect correlated well with the segment length of dyskinesis/akinesis. The values for AS/CD in patients with successful reperfusion significantly and progressively decreased until day 14; 1.00 +/- 0.02 at day 1, 0.93 +/- 0.11 at day 2 (p < 0.05 versus day 1), 0.84 +/- 0.16 at day 3 (p < 0.05 versus day 2), 0.80 +/- 0.13 at day 7 (p < 0.01 versus day 2), 0.73 +/- 0.10 at day 14, and 0.72 +/- 0.10 at day 28. Greater improvement in function was obtained in patients reperfused within 4 hours than in those reperfused at > or = 4 hours (AS/CD at day 28, 0.64 +/- 0.12 versus 0.75 +/- 0.09, p < 0.05). CONCLUSIONS: Thus, a significant amount of myocardium, an average of 28% in segment length of the risk area, is salvaged in patients with reperfused anterior myocardial infarction. Major functional improvement seems to be achieved within 14 days of reflow. PMID- 8425285 TI - Radiofrequency catheter ablation of ventricular tachycardia in patients with coronary artery disease. AB - BACKGROUND: Radiofrequency (RF) ablation of idiopathic ventricular tachycardia (VT) has been demonstrated to be highly efficacious, but the efficacy of RF ablation of VT in patients with coronary artery disease has been unknown. Therefore, the purpose of this study was to determine the feasibility of RF ablation of VT in patients with coronary artery disease. METHODS AND RESULTS: Fifteen consecutive patients with coronary artery disease and a history of myocardial infarction underwent an attempt at RF ablation of 16 hemodynamically stable monomorphic VTs that had been documented clinically on a 12-lead ECG and that had not been successfully managed by pharmacological or device therapy. One VT was incessant, five occurred more than 25 times, and the remainder occurred two to 20 times. An additional four VTs that had not been documented clinically also were targeted for ablation. The mean age of the patients was 68 +/- 7 years (+/- SD), and their mean left ventricular ejection fraction was 0.27 +/- 0.08. The mean cycle length of the 20 VTs targeted for ablation was 438 +/- 82 msec. Ablation sites were selected based on endocardial activation mapping, pace mapping, identification of an isolated mid-diastolic potential, or concealed entrainment. Sixteen of the 20 VTs (80%) were successfully ablated in 11 of 15 patients (73%), using a mean of 4.2 +/- 3 applications of RF energy, and no recurrences of the ablated VTs occurred during 9.1 +/- 3.3 months of follow-up. The mean duration of the ablation procedures was 128 +/- 30 minutes. No complications occurred in any of the patients. CONCLUSIONS: The results of this study demonstrate that RF ablation of hemodynamically stable VT is feasible as adjunctive therapy in selected patients with coronary artery disease. PMID- 8425286 TI - Characterization of double potentials during ventricular tachycardia. Studies during transient entrainment. AB - BACKGROUND: Double potentials have been recorded during reentrant tachycardias in animal models. Although they have also been recorded during ventricular tachycardia in humans, their meaning is uncertain. METHODS AND RESULTS: We used transient entrainment as a method to help further understand the meaning of double potentials recorded during ventricular tachycardia in humans. Three patients with ventricular tachycardia (cycle lengths, 500, 450, and 290 msec) were studied. During transient entrainment of ventricular tachycardia (pacing cycle length, 470-260 msec), both double potential deflections were captured at the pacing cycle length. One deflection was captured with a short activation time, and the other deflection was captured with a long activation time. During ventricular pacing, the deflections were associated by the long rather than the short interdeflection interval. At termination of pacing, each double potential deflection was associated with separate but sequential QRS complexes, and each deflection maintained the same electrogram morphology at relatively "slow" overdrive pacing rates. The short interdeflection interval shortened further with faster pacing rates (to less than the ventricular refractory period), making it unlikely that both deflections of the double potential represent active depolarization of the same tissue. In two patients, at a critically rapid pacing rate, one of the double potential deflections changed morphology abruptly, associated with a shortened stimulus-to-double potential time interval (from 520 to 110 msec and from 530 to 240 msec, respectively), indicating a change in the direction of activation that caused that deflection. Interruption of ventricular tachycardia was associated with disappearance of the double potentials at the same recording site. The double potentials did not immediately bracket an area of slow conduction. CONCLUSIONS: These data suggest that double potentials recorded only during ventricular tachycardia represent activation wave fronts on either side of an area of block within a reentrant circuit. Thus, double potentials recorded during ventricular tachycardia in these patients do not appear to represent slow conduction per se but rather appear to represent an area of block at the center of a reentrant circuit around which the reentrant wave front circulates. PMID- 8425287 TI - Response of nonreentrant catecholamine-mediated ventricular tachycardia to endogenous adenosine and acetylcholine. Evidence for myocardial receptor-mediated effects. AB - BACKGROUND: Reentrant ventricular tachycardia (VT) is known to be insensitive to the nucleoside adenosine. However, we have previously identified a form of nonreentrant, catecholamine-mediated VT that can be initiated with rapid pacing, demonstrates cycle length dependence, and is sensitive to exogenous adenosine as well as to the Valsalva maneuver. The mechanism of this tachycardia is thought to be due to a catecholamine-induced, cAMP-mediated increase in intracellular calcium, resulting in delayed afterdepolarizations and triggered activity. The antiarrhythmic effects of exogenous adenosine and Valsalva on this form of VT may be due to receptor-mediated inhibition of adenylate cyclase or to noncardiac receptor-mediated effects, i.e., exogenous adenosine may modulate VT through alterations in autonomic tone by activation of arterial chemoreceptors, and Valsalva has been shown to decrease venous return, resulting in a reduction in cardiac dimensions and myocardial stretch. To clarify this issue and circumvent both autonomic and noncardiac receptor effects, the response of nonreentrant catecholamine-mediated VT to endogenous adenosine and acetylcholine was evaluated. METHODS AND RESULTS: Group 1 (n = 8): Dipyridamole (0.56 mg/kg i.v.), a nucleoside transport blocker that potentiates the effects of endogenous adenosine, reproducibly abolished sustained nonreentrant, nonautomatic, catecholamine-mediated VT in the five patients in whom it was evaluated. VT recurred with the addition of aminophylline, a competitive adenosine A1-receptor antagonist. Edrophonium (10 mg i.v.), a cholinesterase inhibitor that potentiates the effects of acetylcholine at the muscarinic cholinergic receptor, terminated VT in four of four patients, an effect that was reversed by atropine. Group 2 (n = 6): In patients with reentrant VT, dipyridamole and edrophonium had no effect on VT cycle length or duration. Group 3 (n = 4): Adenosine and vagal maneuvers had no effect on catecholamine-mediated VT caused by automaticity in three of four patients tested. In one patient, adenosine transiently suppressed VT (< 5 seconds), after which it spontaneously resumed. CONCLUSIONS: The results of this study further delineate the mechanism of a newly recognized form of clinical VT. It can be identified by termination of the tachycardia in response to activation of the adenosine A1 or muscarinic cholinergic receptor, which results in inhibition of adenylate cyclase. These receptor-mediated effects appear to be specific for identifying nonreentrant, nonautomatic, catecholamine-mediated VT. PMID- 8425288 TI - Hemodynamic and autonomic nervous system responses to mixed meal ingestion in healthy young and old subjects and dysautonomic patients with postprandial hypotension. AB - BACKGROUND: Although postprandial hypotension is a common cause of falls and syncope in elderly persons and in patients with autonomic insufficiency, the pathophysiology of this disorder remains unknown. METHODS AND RESULTS: We examined the hemodynamic, splanchnic blood pool, plasma norepinephrine (NE), and heart rate (HR) power spectra responses to a standardized 400-kcal mixed meal in 11 healthy young (age, 26 +/- 5 years) and nine healthy elderly (age, 80 +/- 5 years) subjects and 10 dysautonomic patients with symptomatic postprandial hypotension (age, 65 +/- 16 years). Cardiac and splanchnic blood pools were determined noninvasively by radionuclide scans, and forearm vascular resistance was determined using venous occlusion plethysmography. In healthy young and old subjects, splanchnic blood volume increased, but supine blood pressure remained unchanged after the meal. In both groups, HR increased and systemic vascular resistance remained stable. Forearm vascular resistance and cardiac index increased after the meal in elderly subjects, whereas these responses were highly variable and of smaller magnitude in the young. Young subjects demonstrated postprandial increases in low-frequency HR spectral power, representing cardiac sympatho-excitation, but plasma NE remained unchanged. In elderly subjects, plasma NE increased after the meal but without changes in the HR power spectrum. Patients with dysautonomia had a large postprandial decline in blood pressure associated with no change in forearm vascular resistance, a fall in systemic vascular resistance, and reduction in left ventricular end diastolic volume index. HR increased in these patients but without changes in plasma NE or the HR power spectrum. CONCLUSIONS: 1) In healthy elderly subjects, the maintenance of blood pressure homeostasis after food ingestion is associated with an increase in HR, forearm vascular resistance, cardiac index, and plasma NE. In both young and old, systemic vascular resistance is maintained. 2) Dysautonomic patients with postprandial hypotension fail to maintain systemic vascular resistance after a meal. This impairment in vascular response to meal ingestion may underlie the development of postprandial hypotension. 3) The measurement of mean HR or plasma NE does not adequately characterize autonomic cardiac control. Power spectral analysis suggests an impairment in the postprandial autonomic modulation of HR in healthy elderly and dysautonomic subjects, possibly predisposing to hypotension when vascular compensation is inadequate. PMID- 8425289 TI - Hypertrophic cardiomyopathy mutation is expressed in messenger RNA of skeletal as well as cardiac muscle. AB - BACKGROUND: The beta-myosin heavy chain (beta-MHC) gene has been identified as a major locus for familial hypertrophic cardiomyopathy (FHCM). We recently showed that one of the common mutations associated with FHCM is expressed in the cardiac muscle messenger RNA (mRNA) of an affected individual. Since beta-MHC is a major sarcomeric protein of cardiac and skeletal muscle, studies were performed to determine whether the mutation is also expressed in skeletal muscle. METHODS AND RESULTS: Biopsies were obtained of skeletal muscle (biceps brachii) from a proband with FHCM known to have the missense mutation in exon 13 of the beta-MHC gene. RNA was extracted from skeletal muscle and lymphocytes by the RNAzol method. First-strand complementary DNA was synthesized by reverse transcription using an antisense primer to exon 16. Polymerase chain reaction (PCR) was performed using primers to exons 12 and 14 to amplify the segment encompassing exon 13. The PCR products were digested with Ddel restriction endonuclease. Undigested PCR product in the control and the proband was 321 base-pairs (bp). Ddel digestion of the PCR product from normal skeletal and lymphocytes showed two DNA fragments of 181 and 140 bp as expected, whereas digestion of the PCR product from the proband's skeletal muscle and lymphocytes showed four DNA fragments of 181, 149, 140, and 32 bp due to the mutation in exon 13. This indicates that the mutation in affected individuals is also expressed in the mRNA of skeletal muscle and lymphocytes. CONCLUSIONS: To our knowledge, this is the first documentation of a beta-MHC gene mutation expressed in skeletal muscle. This finding is provocative. Does it impair skeletal muscle function? If so, how? If not, why not? Is the impairment, or lack of it, a clue to the molecular defect of cardiac muscle? Furthermore, skeletal muscle provides a readily accessible source of mRNA for expression studies and for purification of the beta-MHC protein, which is probably essential to future investigation designed to unravel the molecular basis of this disorder. PMID- 8425290 TI - Chronic changes in skeletal muscle histology and function in peripheral arterial disease. AB - BACKGROUND: Peripheral arterial disease (PAD) is associated with an impairment in exercise performance and muscle function that is not fully explained by the reduced leg blood flow during exercise. This study characterized the effects of PAD on muscle function, histology, and metabolism. METHODS AND RESULTS: Twenty six patients with PAD and six age-matched control subjects were studied. Ten of the PAD patients had unilateral disease, which permitted paired comparisons between their diseased and nonsymptomatic legs. All PAD patients had a lower peak treadmill walking time and peak oxygen consumption than controls. Vascular disease (diseased leg in unilateral patients and the most severely diseased leg in bilateral patients) was associated with decreased calf muscle strength compared with control values. In patients with unilateral disease, the diseased legs had a greater percentage of angular fibers (indicating chronic denervation) and a decreased type II fiber cross-sectional area (expressed as percent of total fiber area) compared with the nonsymptomatic, or control, legs. In diseased legs, gastrocnemius muscle strength was correlated with the total calf cross-sectional area (r = 0.78, p < 0.05) and type II fiber cross-sectional area (r = 0.63, p < 0.05). Activities of citrate synthase, phosphofructokinase, and lactate dehydrogenase in all 26 PAD patients (most diseased leg) did not differ from control values. Despite a wide range in citrate synthase activity in PAD patients, activity of this enzyme was not correlated with muscle strength or treadmill exercise performance. CONCLUSIONS: In patients with PAD, gastrocnemius muscle weakness is associated with muscle fiber denervation and a decreased type II fiber cross-sectional area. In contrast, the PAD patients displayed substantial heterogeneity in muscle enzyme activities that was not associated with exercise performance. Denervation and type II fiber atrophy may contribute to the muscle dysfunction in patients with PAD and further confirm that the pathophysiology of chronic PAD extends beyond arterial obstruction. PMID- 8425291 TI - Impairment of endothelium-dependent pulmonary artery relaxation in children with congenital heart disease and abnormal pulmonary hemodynamics. AB - BACKGROUND: Endothelial injury may be an important event in the pathophysiology of pulmonary hypertension. We therefore investigated whether endothelial dysfunction occurs early in children with congenital heart defects who are at risk of developing pulmonary vascular disease. METHODS AND RESULTS: In 25 children aged 3-16 years, we studied the response of the pulmonary circulation to graded infusions of acetylcholine (an endothelium-dependent vasodilator) and nitroprusside (a dilator not dependent on endothelial function). Diameter of a bronchopulmonary segment artery and pulmonary blood flow velocity were measured using quantitative angiography and intra-arterial Doppler catheters in 10 children aged 4-16 years with normal pulmonary hemodynamics (controls), seven children aged 3-12 years with left-to-right shunt lesions resulting in increased pulmonary flow, and eight children aged 3-14 years with established pulmonary vascular disease. In the controls, there was a dose-dependent increase in flow velocity in response to acetylcholine (maximal increase, 93 +/- 7%) and in response to nitroprusside (51 +/- 8%). In contrast, in patients with pulmonary vascular disease, the response of flow velocity to similar doses of acetylcholine (33 +/- 7%, p < 0.01) and nitroprusside (7 +/- 13%, p < 0.01) were impaired. In the patients with high pulmonary flow, there was an impaired response to acetylcholine (46 +/- 9%, p < 0.01), but response to nitroprusside was preserved (42 +/- 8%, p > 0.10), consistent with endothelial dysfunction. Arterial diameter was unchanged during acetylcholine infusion in all subjects and increased only modestly in response to nitroprusside (< or = 10%), indicating that the major site of action of each agent is distal to the segmental pulmonary arteries. CONCLUSIONS: Endothelium-dependent pulmonary artery relaxation can be demonstrated in vivo and is impaired in young patients with increased pulmonary flow secondary to congenital heart disease. This impairment may be an important early event in the pathogenesis of pulmonary vascular disease. PMID- 8425292 TI - Inhaled nitric oxide in congenital heart disease. AB - BACKGROUND: Congenital heart lesions may be complicated by pulmonary arterial smooth muscle hyperplasia, hypertrophy, and hypertension. We assessed whether inhaling low levels of nitric oxide (NO), an endothelium-derived relaxing factor, would produce selective pulmonary vasodilation in pediatric patients with congenital heart disease and pulmonary hypertension. We also compared the pulmonary vasodilator potencies of inhaled NO and oxygen in these patients. METHODS AND RESULTS: In 10 sequentially presenting, spontaneously breathing patients, we determined whether inhaling 20-80 ppm by volume of NO at inspired oxygen concentrations (FIO2) of 0.21-0.3 and 0.9 would reduce the pulmonary vascular resistance index (Rp). We then compared breathing oxygen with inhaling NO. Inhaling 80 ppm NO at FIO2 0.21-0.3 reduced mean pulmonary artery pressure from 48 +/- 19 to 40 +/- 14 mm Hg and Rp from 658 +/- 421 to 491 +/- 417 dyne.sec.cm-5.m-2 (mean +/- SD, both p < 0.05). Increasing the FIO2 to 0.9 without adding NO did not reduce mean pulmonary artery pressure but reduced Rp and increased the ratio of pulmonary to systemic blood flow (Qp/Qs), primarily by increasing Qp (p < 0.05). Breathing 80 ppm NO at FIO2 0.9 reduced mean pulmonary artery pressure and Rp to the lowest levels and increased Qp and Qp/Qs (all p < 0.05). While breathing at FIO2 0.9, inhalation of 40 ppm NO reduced Rp (p < 0.05); the maximum reduction of Rp occurred while breathing 80 ppm NO. Inhaling 80 ppm NO at FIO2 0.21-0.9 did not alter mean aortic pressure or systemic vascular resistance. Methemoglobin levels were unchanged by breathing up to 80 ppm NO for 30 minutes. CONCLUSIONS: Inhaled NO is a potent and selective pulmonary vasodilator in pediatric patients with congenital heart disease complicated by pulmonary artery hypertension. Inhaling low levels of NO may provide an important and safe means for evaluating the pulmonary vasodilatory capacity of patients with congenital heart disease without producing systemic vasodilation. PMID- 8425293 TI - Different secretion patterns of atrial natriuretic peptide and brain natriuretic peptide in patients with congestive heart failure. AB - BACKGROUND: The plasma levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) are increased in relation to the severity of congestive heart failure (CHF). This study was designed to examine whether the secretion patterns of ANP and BNP vary with underlying cardiac disorders of CHF with different degrees of overload in atria and ventricles. METHODS AND RESULTS: We measured plasma levels of ANP and BNP in the aorta in 20 patients with mitral stenosis (MS) in whom atria are mainly overloaded, 30 patients with dilated cardiomyopathy (DCM) in whom both atria and ventricles are overloaded, and 20 control subjects during cardiac catheterization. Pulmonary capillary wedge pressure (PCWP) was significantly higher in the MS and DCM groups (16.7 +/- 4.7 mm Hg and 15.1 +/- 7.7 mm Hg, respectively) than in the control group (7.2 +/- 1.1 mm Hg, p < 0.01), whereas there was no significant difference between the MS and DCM groups. Left ventricular end-diastolic pressure (LVEDP) was significantly higher in the DCM group than in the MS group (16.4 +/- 7.8 mm Hg versus 7.6 +/- 2.0 mm Hg, p < 0.01), and the level was comparable between the MS and control groups (7.6 +/- 2.0 mm Hg versus 6.8 +/- 1.2 mm Hg, p = NS). The plasma ANP level was significantly higher in the MS and DCM groups (356 +/- 169 pg/ml and 331 +/- 323 pg/ml, respectively) than in the control group (98 +/- 41 pg/ml, p < 0.01), whereas there was no significant difference between the MS and DCM groups. The plasma BNP level was significantly higher in the DCM group than in the MS group (333 +/- 405 pg/ml versus 147 +/- 54 pg/ml, p < 0.01), and the level was significantly higher in the MS group than in the control group (147 +/- 54 pg/ml versus < 10 pg/ml, p < 0.01). The plasma levels of ANP and BNP had a highly positive correlation with PCWP in the DCM group (p < 0.01). On the other hand, in the MS group, the plasma ANP level had a highly significant correlation with PCWP (p < 0.01) but the plasma BNP level did not. CONCLUSIONS: We conclude that plasma levels of BNP mainly reflect the degree of ventricular overload and that the secretion patterns of ANP and BNP vary with underlying cardiac disorders of CHF with different degrees of overload in atria and ventricles. PMID- 8425294 TI - Exertional fatigue due to skeletal muscle dysfunction in patients with heart failure. AB - BACKGROUND: Exertional fatigue in patients with chronic heart failure is usually attributed to skeletal muscle underperfusion. Recently, skeletal muscle atrophy, abnormal muscle metabolic responses, and reduced muscle enzyme levels have been noted in such patients, raising the possibility that some patients may develop muscle fatigue due to intrinsic muscle abnormalities. The present study was undertaken to determine if a subpopulation of patients with heart failure develops exertional fatigue due to skeletal muscle dysfunction rather than to reduced muscle flow. METHODS AND RESULTS: All exercise hemodynamic studies performed in our laboratory on patients with heart failure were reviewed to identify those who exhibited peak exercise VO2 levels < or = 18 ml.min-1 x kg-1 due to leg fatigue and who underwent insertion of a Swan-Ganz catheter and leg blood flow catheter. Thirty-four patients were identified. Six normal subjects were also studied to define normal leg flow and femoral venous lactate responses to exercise. Patients with peak exercise leg flow levels within the normal mean flow level +/- 2 SEM were considered to have normal skeletal muscle flow during exercise. Nine of the 34 patients with heart failure were found to have normal leg blood flow during exercise. All of these patients terminated exercise due to leg fatigue, and all exhibited abnormal increases in femoral venous lactate concentrations (slope of work load versus femoral venous lactate: normal, 0.33 +/ 0.07 mg/W; heart failure with normal flow, 0.81 +/- 0.08 mg/W; p < 0.002). There was no significant difference between patients with normal leg flows and those with reduced flow in age, ejection fraction, and resting hemodynamic measurements. However, patients with normal flows exhibited more normal cardiac output responses to exercise and tended to have higher peak exercise VO2 (14.1 +/ 0.9 versus 11.5 +/- 0.7 ml.min-1 x kg-1, p < 0.05). CONCLUSIONS: A substantial percentage of patients with chronic heart failure develop exertional fatigue due to skeletal muscle dysfunction rather than to reduced skeletal muscle blood flow. In such patients, therapeutic interventions probably should be directed at improving the skeletal muscle abnormalities rather than at improving skeletal muscle flow. PMID- 8425295 TI - Echocardiographic correlates of left ventricular structure among 844 mildly hypertensive men and women in the Treatment of Mild Hypertension Study (TOMHS). AB - BACKGROUND: Echocardiography provides a noninvasive means of assessing left ventricular (LV) structure and evidence of LV wall remodeling in hypertensive persons. The relation of demographic, biological, and other factors with LV structure can be assessed. METHODS AND RESULTS: LV structure was assessed by M mode echocardiograms for 511 men and 333 women with mild hypertension (average blood pressure, 140/91 mm Hg). Measurements of LV wall thicknesses and internal dimensions were made, and estimates of LV mass indexes and other derivations of structure were calculated. LV hypertrophy criteria were based on previously reported echocardiographic population studies of normal subjects. These measures were compared by age, sex, race, body mass index, systolic blood pressure, antihypertensive drug use, physical activity, alcohol intake, cigarette smoking, and urinary sodium excretion. Despite virtual absence of ECG-determined LV hypertrophy, 13% of men and 20% of women had echocardiographically determined LV hypertrophy indexed by body surface area (g/m2), and 24% of men and 45% of women had LV hypertrophy indexed by height (g/m). Black participants had slightly higher mean levels of wall thickness than nonblack participants but similar LV mass. Systolic blood pressure and urinary sodium excretion were significantly and independently associated with LV mass index and LV hypertrophy using both g/m2 and g/m. Body mass index was significantly related to LV mass index and LV hypertrophy using g/m. Smoking was significantly associated with LV mass index, i.e., using continuous measurement but not using the dichotomy for LV hypertrophy. CONCLUSIONS: This study of a large population of men and women with mild primary hypertension, largely without ECG evidence of LV hypertrophy, showed a substantial percentage of participants with echocardiographically determined LV hypertrophy. LV mass indexes correlated positively with systolic blood pressure, body mass index, urinary sodium excretion, and smoking. PMID- 8425296 TI - Radiofrequency endocardial catheter ablation of accessory atrioventricular pathway atrial insertion sites. AB - BACKGROUND: High rates of success using radiofrequency ablation energy have rapidly transformed catheter ablation from an investigational procedure to the nonpharmacological therapy of choice for symptomatic Wolff-Parkinson-White syndrome. Prior studies of radiofrequency accessory pathway ablation were based on a ventricular approach. Risks associated with prolonged arterial catheter manipulation, retrograde left ventricular catheterization, and production of ventricular lesions required for successful ventricular insertion ablation can be avoided using atrial insertion ablation procedures. The purpose of the present study was to define the safety and efficacy of accessory pathway ablation using radiofrequency energy delivered solely to accessory atrioventricular pathway atrial insertion sites. METHODS AND RESULTS: One hundred fourteen patients with accessory pathway-mediated tachycardia underwent attempted radiofrequency current ablation at the accessory pathway atrial insertion site. All catheters were introduced transvenously. Left-sided accessory pathways were approached using transseptal left atrial catheterization techniques. Retrograde localization of the atrial insertion site during reentrant tachycardia was characterized by 40 +/ 15-msec local ventriculoatrial and 79 +/- 17-msec surface QRS to local atrial electrogram intervals. Presumed accessory pathway potentials were present in only 30% of ablation site electrograms. Successful ablation required 6.2 +/- 5.3 radiofrequency energy applications. Cumulative energy dose required for success was 2,341 +/- 2,233 J. There were no complications associated with transseptal catheterization. Energy delivery to accessory pathway atrial insertion sites was associated with non-life-threatening complications in two patients. Recurrent conduction requiring repeat ablation occurred in 10 of 115 (9%) successfully ablated accessory pathways, all within 1 month of the ablation procedure. After 21.2 +/- 4.6 months of follow-up, 108 of 114 (95%) patients are asymptomatic and without evidence of accessory pathway conduction. CONCLUSIONS: The atrial insertion approach to accessory pathway ablation is safe and highly effective. This approach compares favorably with the retrograde ventricular insertion ablation technique. Atrial insertion ablation eliminates the need to produce ventricular lesions and avoids the risks of prolonged arterial catheter manipulation and retrograde left ventricular catheterization. PMID- 8425297 TI - Augmentation of coronary blood flow by intra-aortic balloon pumping in patients after coronary angioplasty. AB - BACKGROUND: Controversy exists regarding the ability of intra-aortic balloon pumping to increase coronary blood flow in patients with obstructive coronary artery disease. To assess the effects of intra-aortic balloon pumping on coronary hemodynamics, we measured coronary blood flow velocity with a 0.018-in. Doppler tipped angioplasty guide wire in 15 patients who received an intra-aortic balloon pump for typical clinical indications. METHODS AND RESULTS: Intra-aortic balloon pumping augmented diastolic pressure 83 +/- 35%. In nine patients before angioplasty, peak diastolic coronary flow velocity beyond the stenosis (mean diameter narrowing, 95 +/- 7%) was 5.3 +/- 9.6 cm/sec and was unaffected by intra aortic balloon pumping. After angioplasty, the improved coronary luminal diameter narrowing (n = 12; mean narrowing, 18 +/- 12%) was associated with increased distal diastolic flow velocity integral and peak diastolic and mean velocities (13.3 +/- 8.4 units: 36.4 +/- 18.3 and 24.0 +/- 11.4 cm/sec, respectively; all p < 0.01 versus before angioplasty), which were further augmented (36 +/- 37%, 54 +/- 49%, and 26 +/- 17%, respectively; all p < 0.01) with intra-aortic balloon pumping. Intra-aortic balloon pumping did not significantly increase the distal systolic velocity integral (10 +/- 59%) or peak systolic velocity (3 +/- 33%). Similar degrees of balloon pump augmentation of distal coronary flow velocity values were observed in five angiographically normal reference arteries in four patients. CONCLUSIONS: These data demonstrate lack of significant flow improvement beyond most critical stenoses with intra-aortic balloon pumping and the unequivocal restoration and intra-aortic balloon pump-mediated augmentation of both proximal and distal coronary blood flow velocities after amelioration of severe coronary obstructions in patients after successful coronary angioplasty. PMID- 8425298 TI - Direct comparison of [13N]ammonia and [15O]water estimates of perfusion with quantification of regional myocardial blood flow by microspheres. AB - BACKGROUND: Both [13N]ammonia and [15O]water have been used to quantify myocardial blood flow with positron emission tomography using appropriate tracer kinetic models. A direct comparison of the two tracers with radioactive microspheres has not been performed in the same experimental preparation. METHODS AND RESULTS: The two tracers have been tested for myocardial blood flow quantification in closed-chest dogs with circumflex coronary stenosis or permanent occlusion at rest and during adenosine-induced hyperemia. [13N]ammonia- and [15O]water-derived myocardial blood flow values have been compared with radiolabeled microspheres. Validation studies consisted of simultaneous measurements of blood flow with positron emission tomography and microspheres over a wide range of flow values. Blood pool and regional tissue activity curves were fitted with a three-compartment model for [13N]ammonia with and without arterial metabolite correction and with a single-tissue-compartment model for [15O]water. A correction for finite-resolution effect before the fit was also applied. In large regions of interest (5 cm3), a good correlation between the microsphere method and [13N]ammonia (with metabolite correction) was obtained (y = 3 + 0.78x, r = 0.94). The correlation with microspheres was slightly better with [15O]water (y = -3 + 0.89x, r = 0.97). Similar correlations were achieved in smaller regions of interest (1 cm3) as well as in akinetic segments and in central infarct regions. CONCLUSIONS: Positron emission tomography with appropriate tracer kinetic models using [13N]ammonia and [15O]water provides an accurate quantitative method for measuring regional myocardial blood flow over a wide range of flow values in normally contracting or akinetic canine myocardium in the absence and in the presence of infarction. PMID- 8425299 TI - Acute effects of nitroglycerin depend on both plasma and intracellular sulfhydryl compound levels in vivo. Effect of agents with different sulfhydryl-modulating properties. AB - BACKGROUND: Changes in sulfhydryl (SH) compound availability may alter the hemodynamic effect of nitroglycerin (NTG). Data on the relation between NTG effect and thiol levels are, however, limited to in vitro experiments. The present study investigates how intracellular and extracellular changes in SH group concentrations (cysteine and glutathione [GSH]) affect the responsiveness to NTG in vivo. METHODS AND RESULTS: GSH and cysteine levels in plasma, vena cava, and aorta were measured after administration of N-acetylserine (placebo, n = 6), N-acetylcysteine (NAC, extracellular and intracellular SH donor, n = 6), oxothiazolidine (OXO, intracellular SH donor, n = 6), buthionine sulfoximine (BSO, intracellular GSH-depleting agent, n = 6), BSO+NAC (n = 6), and BSO+OXO (n = 6) in chronically catheterized conscious rats. In addition, the effect of 2.5 mg NTG/kg i.v. on mean arterial pressure (MAP) was determined before and after the same treatment. NAC (5 mmol/kg i.v. for 2 hours) significantly (p < 0.05) increased extracellular cysteine and GSH levels and potentiated the hypotensive effect of NTG (from 26 +/- 3 to 31 +/- 4 mm Hg [mean +/- SEM], p < 0.05). OXO (5 mmol.kg-1 x hr-1 i.v. for 2 hours) significantly increased intracellular cysteine and GSH levels but had no effect on NTG responsiveness (p > 0.05). BSO (1 g i.p. three times within 24 hours) significantly decreased intracellular GSH levels (p < 0.05) and attenuated the effect of NTG (from 28 +/- 3 to 16 +/- 2 mm Hg). CONCLUSIONS: The results suggest that the acute hypotensive effect of NTG in vivo is: 1) increased by high extracellular GSH and/or cysteine levels (NAC), 2) decreased by low intracellular GSH levels (BSO), and 3) unaffected by high intracellular levels of cysteine and GSH (OXO). PMID- 8425300 TI - Flecainide-induced arrhythmia in canine ventricular epicardium. Phase 2 reentry? AB - BACKGROUND: We recently reported that sodium channel block can produce opposite effects on action potential duration (APD) and refractoriness in epicardial versus endocardial tissues of the canine ventricle. In addition, strong sodium channel current inhibition was found to cause loss of the action potential dome in epicardium but not endocardium, thus inducing a marked dispersion of repolarization and refractoriness between epicardium and endocardium as well as among neighboring epicardial sites. The marked heterogeneity that evolves under these conditions provides a substrate for the development of arrhythmias. Flecainide was found to induce extrasystolic activity more readily than other sodium blockers. The present study contrasts the electrophysiological actions of flecainide in canine ventricular epicardium and endocardium and examines the characteristics of flecainide-induced arrhythmias in epicardial sheets of canine ventricle. METHODS AND RESULTS: Standard microelectrode techniques were used. Flecainide (10-20 microM) produced either prolongation or marked abbreviation of APD in epicardium but only minor changes in the APD of endocardium. Marked abbreviation of APD in epicardium was due to loss of the action potential dome (plateau phase). Arrhythmias displaying characteristics of reentry could be readily induced in flecainide-treated preparations either by increasing the stimulation rate or by introduction of extrastimuli. Flecainide-induced slowing of conduction, more accentuated at the faster stimulation rates, appeared to act synergistically with the drug-induced dispersion of repolarization to generate reentry in these relatively small sheets of epicardium. 4-Aminopyridine, a transient outward current (Ito) blocker, reversed the flecainide-induced marked abbreviation of APD in epicardium and abolished reentrant activity in all cases. Flecainide failed to induce reentry in preparations pretreated with 4 aminopyridine. CONCLUSIONS: Our data suggest that the presence of a prominent Ito in epicardium contributes the development of marked electrical heterogeneity in the ventricle after exposure to flecainide. Flecainide-induced dispersion of repolarization, especially when accompanied by prominent conduction delays, results in extrasystolic activity via a mechanism that we have termed "phase 2 reentry." Our results also suggest a role for Ito blockers in the treatment of reentrant arrhythmias. PMID- 8425301 TI - Role of coronary artery spasm in progression of organic coronary stenosis and acute myocardial infarction in a swine model. Importance of mode of onset and duration of coronary artery spasm. AB - BACKGROUND: Coronary spasm may play an important role in progression of organic coronary stenosis and myocardial infarction, but the mechanisms responsible for these complications are not known. This study aimed to examine whether the mode of onset and the duration of coronary spasm influenced progression of organic coronary stenosis and acute myocardial infarction in a swine model of coronary spasm. METHODS AND RESULTS: Gottingen miniature pigs were subjected to cholesterol feeding, balloon-induced coronary arterial denudation, and x-ray irradiation. Five months later, coronary spasm was induced by intracoronary injection of serotonin. In 10 pigs, coronary spasm was provoked abruptly and maintained for 25 minutes by five repeated intracoronary injections of serotonin (10 micrograms/kg) every 5 minutes (group A, abrupt onset and short duration). In group B, coronary spasm was provoked gradually by intracoronary injections of serotonin at graded doses of 0.1, 0.3, and 0.6 microgram/kg every 5 minutes and was then maintained for 25 minutes in four pigs (group B1, gradual onset and short duration) and for 120 minutes in six pigs (group B2, gradual onset and long duration) by repeated intracoronary injections of serotonin (10 micrograms/kg) every 5 minutes. Intramural hemorrhage was noted histologically at the spastic site more frequently in group A with abrupt onset (nine of 10 pigs) than in group B with gradual onset (two of 10 pigs) (p < 0.01). Progression of organic coronary stenosis due to intramural hemorrhage was noted in seven pigs (six pigs in group A and one pig in group B), including three cases of total coronary occlusion. Evidence for the evolution of acute myocardial infarction (serial ECG findings, left ventriculograms, and histological findings) was noted in one pig (7%) of group A or B1 with short duration and in five of six pigs (83%) in group B2 with long duration (p < 0.01 versus group A and B1). CONCLUSIONS: These results indicate that: 1) intramural hemorrhage was frequently induced by coronary spasm of abrupt but not of gradual onset, 2) intramural hemorrhage resulted in acute progression of coronary stenosis and sometimes resulted in persistent total coronary occlusion leading to acute myocardial infarction, and 3) prolonged coronary spasm resulted in acute myocardial infarction without progression of organic coronary stenosis. PMID- 8425302 TI - Effect of aspirin on coronary collateral blood flow. AB - BACKGROUND: Although aspirin exerts beneficial antiplatelet activity in patients with coronary artery disease, cyclooxygenase blockade produced by aspirin causes a potentially deleterious effect by interrupting endothelial production of prostacyclin. Collateral vessels that develop in response to coronary occlusion display prominent endothelial cell proliferation and undergo vasoconstriction in response to indomethacin. This study was performed to test the hypothesis that cyclooxygenase blockade with aspirin would cause constriction of coronary collateral vessels and that such vasoconstriction would be reversed with nitroglycerin. METHODS AND RESULTS: Collateral vessel growth was induced by embolic occlusion of the left anterior descending coronary artery in dogs. Four to 6 months later, coronary collateral flow was measured as retrograde flow from the cannulated collateral-dependent artery. Aspirin (1 mg/kg i.v.) caused 70 +/- 8% blockade of the increase in coronary blood flow produced by intra-arterial arachidonic acid and decreased retrograde flow from 37 +/- 7 to 28 +/- 7 ml/min (p < 0.03). Increasing the dose of aspirin to 15 mg/kg i.v. caused 91 +/- 3% blockade of the response to arachidonic acid and further decreased retrograde flow to 21 +/- 4 ml/min (p < 0.01). After aspirin administration, nitroglycerin (150 micrograms/min i.c.) reversed the collateral constriction and increased retrograde flow to 37 +/- 10 ml/min (p < 0.01). CONCLUSIONS: These data suggest that products of cyclooxygenase metabolism cause tonic vasodilation of well developed coronary collateral vessels. Blockade of cyclooxygenase with even low dose aspirin caused collateral vessel constriction with a decrease in collateral blood flow. However, nitroglycerin was able to fully reverse aspirin-induced collateral vasoconstriction and restore flow to the control level. PMID- 8425303 TI - SIN-1 reduces platelet adhesion and platelet thrombus formation in a porcine model of balloon angioplasty. AB - BACKGROUND: Nitric oxide inhibits platelet adhesion and platelet aggregation in vivo. In this study, we investigated the effects of the nitric oxide donor SIN-1 on platelet adhesion and platelet-thrombus formation following experimental angioplasty. METHODS AND RESULTS: Bilateral carotid angioplasty was performed in 20 anesthetized pigs. Animals received either SIN-1 (3-morpholino-sydnonimine; 10 micrograms/kg/min; n = 8) or placebo (n = 8) before and during angioplasty. An additional control group of pigs received trimetaphan (n = 4), which induced hemodynamic changes similar to those that followed treatment with SIN-1. Platelet deposition was quantified by the injection of autologous 111In-labeled platelets. SIN-1 reduced platelet deposition after deep arterial injury compared with placebo (mean +/- SEM, 10.870 +/- 2.415 versus 40.326 +/- 9.889 platelets x 10(6)/cm2, p < 0.05). SIN-1 reduced platelet adhesion after superficial injury compared with both placebo and trimetaphan (2.231 +/- 0.333 versus 5.278 +/- 0.606 versus 5.022 +/- 1.136 platelets x 10(6)/cm2, respectively; p < 0.005). Scanning electron microscopy confirmed that platelets were deposited in the form of an adherent monolayer following superficial endothelial denudation and were reduced in number following treatment with SIN-1. The effects of SIN-1 on platelet function were associated with a significant increase in platelet cyclic GMP concentration from baseline (3.15 +/- 0.88 versus 1.58 +/- 0.73 pmol/10(9) platelets, p < 0.005). CONCLUSIONS: SIN-1 reduces platelet adhesion and platelet thrombus formation following experimental angioplasty. The antiadhesive effects of SIN-1 are independent of changes in systemic hemodynamics. These results imply that the administration of a nitric oxide donor may prove effective in modifying the pathophysiological response to angioplasty injury. PMID- 8425304 TI - Myocardial hypertrophy in the ischemic zone induced by exercise in rats after coronary reperfusion. AB - BACKGROUND: Factors influencing left ventricular (LV) remodeling after coronary artery reperfusion, including adaptive changes in the infarcted region and the role of exercise, have not been well defined. The common application of early reperfusion by thrombolysis after acute myocardial infarction lends potential significance to such remodeling, and a rat model with 45 minutes of regional ischemia followed by reperfusion was developed to study these events. We postulated that the effects of reperfusion in altering LV morphology would be further modified by exercise training, including induction of hypertrophic changes in the outer region of the nontransmural infarction. METHODS AND RESULTS: Female Sprague-Dawley rats were subjected either to 15 minutes of left anterior descending coronary artery occlusion followed by reperfusion or to sham operation, and at 5 days after the operation, animals were randomly assigned to sedentary conditions or to 3 weeks of swimming exercise. Animals completing the experiment included a reperfused sedentary group (n = 21), a reperfused exercised group (n = 20), a sham-operated group (n = 10), and a sham-operated group subjected to exercise (sham exercised group, n = 9). In addition, in seven rats, myocardial infarction was produced by permanent coronary occlusion, and the animals remained sedentary (permanent occlusion group). In each group, the morphology of the noninfarcted (septal) and the infarcted (anterolateral) regions of the left ventricle was examined 26 days after surgery from midventricular transverse sections 25 microns thick taken after perfusion fixation of the heart at an aortic pressure of 60 mm Hg and an LV cavity pressure of 10 mm Hg. Compared with the permanent occlusion group, LV cavity area in the sedentary reperfused group was smaller (33.5 versus 53.2 mm2, p < 0.001), the infarcted wall was thicker (1.36 versus 0.53 mm, p < 0.001), and the septal wall also was thicker (1.95 versus 1.62 mm, p < 0.05), whereas compared with the sham-operated group, the LV cavity area was increased, and infarcted wall thickness was reduced (both p < 0.01). Reperfusion resulted in less transmurality of infarction compared with permanent occlusion (38.3% versus 69.5%, p < 0.001), with increased subepicardial area in the infarcted zone after reperfusion (7.5 versus 1.9 mm2, p < 0.001). In the reperfused exercised group, transmurality was further decreased compared with the reperfused sedentary group (31.5% versus 38.3%, p < 0.05), and the viable subepicardial area of the infarct zone increased by 32%. CONCLUSIONS: Important remodeling of global and regional LV morphology was evident at 26 days in sedentary rats after 45 minutes of coronary occlusion with reperfusion compared with rats with permanent coronary occlusion, with reduced infarct transmurality and less LV dilation in the reperfused group. Exercise after reperfusion further affected ventricular remodeling by causing hypertrophy with increased wall thickness of the surviving subepicardium of the infarcted zone. PMID- 8425305 TI - Comparative study on the proarrhythmic effects of some antiarrhythmic agents. AB - BACKGROUND: A main side effect of antiarrhythmic drug therapy is the tendency of these drugs to promote arrhythmia within the therapeutic concentration range, i.e., the proarrhythmic activity of these drugs. However, a model for in vitro assessment, quantification, and comparison of proarrhythmic drug activities was still lacking, and only sparse data were available. METHODS AND RESULTS: To analyze the arrhythmogenic risk of common antiarrhythmic drugs in a quantitative and comparative manner, isolated perfused rabbit hearts were treated with increasing concentrations of antiarrhythmic drugs corresponding to low, medium, and high therapeutic concentrations. For analysis of the epicardial activation process, an epicardial mapping (256 unipolar leads) was performed. For each electrode, the activation time was determined. From these data, the origins of epicardial activation ("breakthrough points" [BTP]) were determined. At each electrode, an activation vector (VEC) was calculated giving direction and velocity of the local excitation wave. The beat similarity of various heartbeats (under treatment) compared with control was evaluated by determination of the percentage of identical BTPs (deviation < or = 1 mm) and of similar VECs (deviation < or = 5 degrees). BTP and VEC were reduced by all antiarrhythmic agents tested (propafenone = flecainide > quinidine > ajmaline > procainamide > disopyramide > mexiletine = lidocaine > sotalol), indicating a more or less pronounced disturbance of the epicardial activation process. Treatment with propafenone, quinidine, and disopyramide and to a lesser extent sotalol prolonged the activation-recovery interval (ARI). ARI dispersion was greatly enhanced by flecainide and was reduced by sotalol. In addition, it could be shown that propranolol is able to reduce the proarrhythmic action of flecainide. This effect seemed to be due to a reduction of the flecainide-induced increase in ARI dispersion. CONCLUSIONS: From the results of our study, we propose the following rank order of the arrhythmogenic risk: flecainide > propafenone > quinidine > ajmaline > disopyramide > procainamide > mexiletine, lidocaine > sotalol. Moreover, we conclude that propranolol given additionally may be helpful in reducing the proarrhythmic risk of flecainide. PMID- 8425306 TI - Deficient neuroendocrine regulation of the cardiovascular system with advancing age in healthy humans. PMID- 8425307 TI - The challenge of further reducing cardiac mortality in the thrombolytic era. PMID- 8425308 TI - Which pharmacological stress is optimal? A technique-dependent choice. PMID- 8425309 TI - Catheter ablation of ventricular tachycardia related to coronary heart disease. Defining the target. PMID- 8425310 TI - Elimination of atrioventricular nodal reentrant tachycardia. PMID- 8425311 TI - Plasma endothelin in chronic heart failure. PMID- 8425312 TI - Principles of access to health care. Access to Health Care Task Force, American Heart Association. PMID- 8425313 TI - Aspirin as a therapeutic agent in cardiovascular disease. Special Writing Group. PMID- 8425314 TI - Long-term follow-up of congenital aortic stenosis, pulmonary stenosis, and ventricular septal defect. PMID- 8425315 TI - Second natural history study of congenital heart defects. PMID- 8425316 TI - Cardiorespiratory responses to exercise of patients with aortic stenosis, pulmonary stenosis, and ventricular septal defect. AB - BACKGROUND: The purpose of this study was to determine exercise tolerance and cardiac response to exercise for a large cohort of adult subjects with aortic stenosis, pulmonary stenosis, or ventricular septal defect participating in the Second Natural History Study of Congenital Heart Defects (NHS-2). METHODS AND RESULTS: Exercise testing was performed on 134 of 235 NHS-2 full participants with aortic stenosis, 195 of 331 NHS-2 full participants with pulmonary stenosis, and 324 of 594 NHS-2 full participants with ventricular septal defect. A Bruce treadmill exercise protocol was used. Mean exercise duration for patients with aortic stenosis, pulmonary stenosis, and ventricular septal defect was 86.5%, 94.2%, and 90.8% of predicted, respectively. For patients with aortic stenosis, there was a direct relation between echocardiographic Doppler maximum transaortic gradient and ST segment change during exercise. ST segment change during exercise was very uncommon for patients with pulmonary stenosis. For patients with ventricular septal defect, there was an association between arrhythmias noted during exercise and the presence of associated aortic insufficiency. CONCLUSIONS: Although exercise duration was well preserved for patients with each of the three defects, exercise tolerance was subnormal. PMID- 8425317 TI - Occurrence risk for congenital heart defects in relatives of patients with aortic stenosis, pulmonary stenosis, or ventricular septal defect. AB - BACKGROUND: Knowledge of the risk of occurrence of congenital heart defects in offspring of individuals with a congenital heart defect is important for genetic counseling and understanding the etiology of congenital heart diseases. METHODS AND RESULTS: A portion of the questionnaire mailed to all patients in the Second Natural History Study of Congenital Heart Defects (NHS-2) addressed marital status, pregnancy, miscarriage, and presence or absence of congenital heart defects or other congenital malformations in first-degree relatives. Offspring were not examined as a part of the study. For male probands with aortic stenosis (AS), three of 251 offspring had congenital heart disease, whereas one of 72 offspring of female probands with AS had congenital heart disease. For patients with pulmonary stenosis (PS), three of 176 offspring of male probands had congenital heart disease, whereas eight of 205 offspring of female probands had congenital heart disease. For male probands with ventricular septal defect (VSD), 10 of 334 offspring had congenital heart defects, and 11 of 384 offspring of female probands with VSD had offspring with congenital heart defects. The prevalence rate for noncardiac congenital anomalies in offspring of probands was 2%. CONCLUSIONS: Based on known congenital heart defects in offspring of probands in the NHS-2, occurrence rates of congenital heart disease in children of subjects with AS, PS, and VSD were 1.2% (confidence interval [CI], 0.34-3.1%), 2.8% (CI, 1.4-5.1%), and 2.9% (CI, 1.8-4.4%), respectively. PMID- 8425318 TI - Bacterial endocarditis in patients with aortic stenosis, pulmonary stenosis, or ventricular septal defect. AB - BACKGROUND: All of the 2,401 patients with aortic stenosis (AS), pulmonary stenosis (PS), or ventricular septal defect (VSD) admitted to the First Natural History Study of Congenital Heart Defects between 1958 and 1965 were eligible for the Second Natural History Study. Most patients with severe defects were managed surgically, and most with mild defects were managed medically. Final examination in the first study was carried out 8 years after admission. METHODS AND RESULTS: For AS, the incidence rate of bacterial endocarditis (BE) was 27.1 per 10,000 person-years. The incidence rate was 15.7 per 10,000 person-years for those managed medically and 40.9 per 10,000 person-years for those managed surgically. Most patients managed surgically had severe AS, and severity was more important to the risk of BE than the method of management. For PS, only one of the 592 patients with PS experienced BE. For VSD, the incidence rate of BE was 14.5 per 10,000 person-years. Size of the VSD was not associated with risk of BE. The risk of BE before closure of the VSD was more than twice that after surgery. CONCLUSIONS: The incidence rate of BE was nearly 35-fold the population-based rate. The increased incidence in patients with AS after valvotomy was a function of severity of the defect and not a function of surgery. Presence of aortic regurgitation in patients with AS did not increase the risk of developing BE. Surgical closure of VSD lowered the risk of BE. PMID- 8425319 TI - Second natural history study of congenital heart defects. Results of treatment of patients with aortic valvar stenosis. AB - BACKGROUND: From 1958 to 1969, 462 patients (mostly children) with aortic stenosis were admitted to the First Natural History Study of Congenital Heart Defects (NHS-1) after cardiac catheterization. Most with gradients < 50 mm Hg were managed medically; most with gradients > or = 80 mm Hg had aortic valvotomy. Of those with gradients of 50-79 mm Hg, some were managed medically, and some were managed surgically. Most had a second cardiac catheterization at the conclusion of NHS-1. More than 15 years have elapsed since NHS-1, and most of the cohort are adults. This report (the Second Natural History Study [NHS-2]) addresses the long-term results of medical and surgical management. METHODS AND RESULTS: Of the original cohort of 462 patients, 440 were alive at completion of NHS-1. New data were obtained on 371 (80.3%) of the original cohort. Probability of 25-year survival was 92.4% for those admitted with gradients < 50 mm Hg and 81.0% for those with admission gradients > or = 50 mm Hg. More than half of the cardiac deaths were sudden and unexpected. Forty percent of patients managed medically during NHS-1 subsequently required surgical management. Almost 40% of operated patients required a second operation. Three percent of the original cohort subsequently had bacterial endocarditis. There was a higher-than-normal prevalence of serious arrhythmias. Of NHS-2 full participants, 92.3% were in New York Heart Association functional class I. Most patients had low Doppler mean gradients. Clinically, 46.6% had aortic valve regurgitation. The final clinical status was excellent in 29.9%, good in 22.8%, fair in 28.6%, and poor in 18.7%. CONCLUSIONS: Patients with gradients < 25 mm Hg can be followed medically; however, progressive stenosis can occur, and approximately 20% will require intervention. If the gradient is > or = 50 mm Hg, there is a risk of serious arrhythmias and, possibly, sudden death. If the gradient is > or = 80 mm Hg, intervention is clearly indicated; as techniques improve, delaying intervention in patients with gradients of 50-79 mm Hg may not be advantageous. Patients with gradients of 25-49 mm Hg can be followed medically with annual evaluation. PMID- 8425320 TI - Second natural history study of congenital heart defects. Results of treatment of patients with pulmonary valvar stenosis. AB - BACKGROUND: From 1958 to 1969, 592 patients (mostly children) with pulmonary stenosis were admitted to the First Natural History Study of Congenital Heart Defects (NHS-1) after cardiac catheterization. Most with gradients < 50 mm Hg were managed medically; most with gradients > or = 80 mm Hg had pulmonary valvotomy. Among these with gradients of 50-79 mm Hg, some were managed medically, and some were managed surgically. Most had a second cardiac catheterization at conclusion of NHS-1. More than 15 years had elapsed since NHS 1, and most of the cohort are adults. This report (Second Natural History Study) addresses the long-term results of medical and surgical management. METHODS AND RESULTS: Of the original cohort of 592 patients, 580 were alive at completion of NHS-1. New data were obtained on 464 (78.4%) of the original cohort. Probability of 25-year survival was 95.7%. Probability of survival was less (80%) in a subgroup of patients entering NHS-1 > 12 years old with cardiomegaly. Less than 20% of patients managed medically during NHS-1 subsequently required a valvotomy. Only 4% of operated patients required a second operation. Bacterial endocarditis occurred rarely. Ninety-seven percent were in New York Heart Association class I. There was a small but higher-than-normal prevalence of serious arrhythmias. Most patients, whether medically or surgically managed, had low Doppler maximum gradients. Final clinical status was excellent or good in 83%. CONCLUSIONS: Probability of survival is similar to that of the general population, and the vast majority of patients are asymptomatic. If valvotomy or valvuloplasty is required in a child, reoperation is rarely necessary. Patients with gradients < 25 mm Hg do not experience an increase in gradient. Patients with a gradient > or = 50 mm Hg should have valvotomy or valvuloplasty. Choice of management of patients with gradients of 40-49 mm Hg remains debatable. PMID- 8425321 TI - Second natural history study of congenital heart defects. Results of treatment of patients with ventricular septal defects. AB - BACKGROUND: From 1958 to 1969, 1,280 patients (mostly children) with ventricular septal defects (VSDs) were admitted to the First Natural History Study of Congenital Heart Defects (NHS-1) after cardiac catheterization. Most with small defects and Eisenmenger's syndrome were managed medically; most with large VSDs were managed surgically. Of those with moderate-size defects, some were managed medically, and some were managed surgically. Most had a second catheterization at the conclusion of NHS-1. More than 15 years have elapsed since NHS-1, and most of the cohort are adults. This report (Second Natural History Study) addresses the long-term results of medical and surgical management. METHODS AND RESULTS: Of an original cohort of 1,280 patients, 1,099 were alive at completion of NHS-1. New data were obtained on 976 (76.3%) of the original cohort. Probability of 25-year survival was 87%, and admission severity was the best predictor of survival. Of the 860 patients managed medically during NHS-1, 245 subsequently required surgical closure of the VSD. Only 5.5% of patients who had surgical closure required a second operation. On follow-up, there was a higher-than-normal prevalence of serious arrhythmias. Bacterial endocarditis occurred rarely. Of patients with small VSDs, 94.1% were in New York Heart Association functional class I. With the exception of those with Eisenmenger's syndrome, most patients had a final clinical status that was excellent or good. CONCLUSIONS: The majority of patients fared well. However, there was a higher-than-normal prevalence of serious arrhythmia and sudden death, including those with small VSDs. PMID- 8425322 TI - Second natural history study of congenital heart defects. Materials and methods. AB - Results of the location and recruitment efforts and comparisons of responses from patients who cooperated at different levels in the Second Natural History Study of Congenital Heart Defects are included because they influenced the choice of analytic methods and are essential to the generalizability of the results to the entire study cohort. Included are examination and data collection protocols (e.g., protocol definitions, test procedures, and data editing), statistical methods (e.g., box plots, survival curves, multivariable models, and rate adjustment), participation results (e.g., proportional odds analysis, mortality, location, recruitment, and full participants, including comparison of questionnaire responses and comparison of questionnaire response and physician history), and a discussion. PMID- 8425323 TI - Second natural history study of congenital heart defects. Quality of life of patients with aortic stenosis, pulmonary stenosis, or ventricular septal defect. AB - BACKGROUND: Quality of life of patients with congenital heart defects is an important aspect of the assessment of outcome of medical and surgical treatment. METHODS AND RESULTS: All participants in the Second Natural History Study of Congenital Heart Defects who completed the study questionnaire were included in this analysis. The questionnaire completed by the patients included inquiries relative to medical history, marital and family life, self-perception of well being, insurability, and employability. The following general observations were apparent. The study cohort reported a self-perception of health status that was similar to that of the general population. Second, 35-40% of patients had not had a cardiac evaluation within the previous 10 years. Third, 16% of the patients had no health insurance, and 35.7% of the patients had no life insurance. For the patients with pulmonary stenosis and ventricular septal defect, the percent who were married was less than that of the corresponding national age- and sex specific population. For patients with aortic stenosis, the percent married was greater than that of the general population for some age groups. The proportion of divorced or separated individuals was similar to that of the general population. For all three defects, the level of educational attainment exceeded the national average. Last, unemployment rates were similar to national averages except for women with aortic stenosis, who had a significantly higher unemployment rate than did age- and sex-matched controls. CONCLUSIONS: Patients initially identified with aortic stenosis, pulmonary stenosis, or ventricular septal defect in 1959-1973 have a quality of life in the mid-1980s similar to that of the general US population. PMID- 8425324 TI - Second natural history study of congenital heart defects. Aortic stenosis: echocardiography. AB - BACKGROUND: Recent advances in the field of echocardiography have made it possible to obtain a complete morphological and hemodynamic assessment of patients with aortic stenosis. Therefore, comprehensive two-dimensional and Doppler examinations were performed on patients with aortic stenosis returning for the Second Natural History Study of Congenital Heart Defects (NHS-2). METHODS AND RESULTS: Two hundred thirty-two patients with aortic stenosis underwent comprehensive two-dimensional and Doppler examinations. Of these, 96 were in the medically treated group, 49 had undergone aortic valve replacement, and 87 had had aortic valvotomy. The valve replacement group had a significantly smaller left ventricular diastolic cavity than did the medically treated group; both had smaller dimensions than the valvotomy group. There was no significant difference in left ventricular wall thickness or fractional shortening among the three groups. The valvotomy group had a significantly higher mean aortic valve gradient than did either the medically treated group or the valve replacement group. The degree of aortic regurgitation was more severe in the valvotomy group than in the other two groups. For all patients, there was a direct relation between the aortic valve gradient and the mean wall thickness and an inverse relation between the aortic valve mean gradient and fractional shortening. The only echocardiographic parameter that correlated with the presence or absence of symptomatology was the fractional shortening. CONCLUSIONS: These observations provide an objective measurement for assessing the status of the aortic valve and left ventricular response in patients with aortic stenosis returning for NHS-2. Limitations of echocardiography in this study are discussed. PMID- 8425325 TI - Second natural history study of congenital heart defects. Pulmonary stenosis: echocardiography. AB - BACKGROUND: Two-dimensional and Doppler echocardiography can provide information about valve morphology, right ventricular size and function, and hemodynamics in patients with pulmonary stenosis. Therefore, two-dimensional and Doppler echocardiographic examinations were performed on patients with pulmonary stenosis returning for the Second Natural History Study of Congenital Heart Defects. METHODS AND RESULTS: Three hundred twenty-five patients with pulmonary stenosis underwent two-dimensional and Doppler echocardiographic examinations. Of these, 115 were in the medically treated group, and 210 had undergone a previous operation. Patients in the valvotomy group had a higher incidence of right ventricular dysfunction and a larger right ventricular diastolic dimension. The valvotomy group had a lower pulmonary valve mean gradient and a lower right ventricular systolic pressure than the medically treated group. For all patients, there was no significant correlation of the echocardiographic variables with the presence or absence of symptoms, reflecting the low incidence of patients with cardiac decompensation. CONCLUSIONS: These observations provide an objective measurement for assessing the status of the pulmonary valve and right ventricular response in patients with pulmonary stenosis. Limitations of echocardiography in this study are discussed. PMID- 8425326 TI - Second natural history study of congenital heart defects. Ventricular septal defect: echocardiography. AB - BACKGROUND: Two-dimensional and Doppler echocardiography can provide structural and hemodynamic information for patients with ventricular septal defects (VSDs). Therefore, two-dimensional and Doppler echocardiographic examinations were performed on patients with VSDs returning for the Second Natural History Study of Congenital Heart Defects. METHODS AND RESULTS: Five hundred fifty-six patients with VSDs underwent two-dimensional and Doppler echocardiographic examinations. Three hundred twenty-four patients were in the medically treated group, and 232 had undergone a previous operation. The location of the VSD was classified in 235 patients. Several Doppler measurements were used to calculate pulmonary artery pressures: tricuspid regurgitation peak systolic velocity, peak systolic velocity of VSD jet, pulmonary regurgitation end-diastolic velocity, and pulmonary artery acceleration time. The two methods that were found to be reliable consisted of the tricuspid regurgitation peak systolic velocity and the pulmonary regurgitation end-diastolic velocity. These measurements were obtained in only 26% of patients at all centers. At one center in which adult and pediatric echocardiography was performed in a single laboratory, these values were obtained in 60% of patients. CONCLUSIONS: Two-dimensional and Doppler echocardiography has the ability to provide a noninvasive method of assessing morphology and hemodynamics in patients with VSDs. However, the reliability and accuracy of hemodynamic measurements are dependent on operator experience. PMID- 8425327 TI - Arrhythmias in patients with valvar aortic stenosis, valvar pulmonary stenosis, and ventricular septal defect. Results of 24-hour ECG monitoring. AB - BACKGROUND: Arrhythmias are a significant source of morbidity and mortality in patients with congenital heart defects. METHODS AND RESULTS: As part of the Second Natural History Study of Congenital Heart Defects (NHS-2), 24-hour ambulatory ECG monitoring was performed for full participants in the study. At least 15.5 hours of monitoring was required for inclusion in the analysis. This was achieved for 755 (90.6%) of the patients. Multiform premature ventricular contractions, ventricular couplets, and ventricular tachycardia were considered "serious arrhythmias." For patients with aortic stenosis, the presence of "serious arrhythmias" was associated with higher left ventricular end-diastolic pressure, presence of aortic regurgitation, male sex, and presence of prior aortic valve replacement. For patients with pulmonary stenosis, only older age on admission to NHS-1 was significantly associated with "serious arrhythmias." For patients with ventricular septal defect, the variables associated with "serious arrhythmias" were different for medically and surgically managed patients. For medically managed patients, higher mean pulmonary artery pressure on admission to NHS-1 and older age on admission to NHS-2 were associated with the presence of serious arrhythmias. For surgically managed patients, higher New York Heart Association functional class and cardiomegaly were associated with serious arrhythmias. CONCLUSIONS: The prevalence of arrhythmias (especially ventricular arrhythmias) was higher for patients with aortic stenosis, pulmonary stenosis, or ventricular septal defect than for an historical control population. "Serious arrhythmias" were most frequently noted in patients with aortic stenosis who also had a higher incidence of sudden death. The prevalence of "serious arrhythmias" was second highest for patients with ventricular septal defect who had the second highest incidence of sudden death. PMID- 8425328 TI - 1992 Proceedings of the Knee Society. PMID- 8425329 TI - Hematologic effects of total knee arthroplasty. A prospective evaluation. AB - In a prospective evaluation of 41 patients (57 knees) treated with index cemented total knee arthroplasty, perioperative blood testing was performed on blood counts, fibrinogen, and fibrin degradation products (FDP). The preoperative platelet count averaged 314/nl (range, 160-502/nl), whereas postoperatively, the count nadired at an average of 173/nl (range, 60-305/nl). Fibrinogen levels increased from 304 mg/dl (range, 170-442 mg/dl) to an average high of 647 mg/dl (range, 317-1018 mg/dl). Patients were also classified according to whether they had been treated with unilateral or bilateral procedures. Postoperatively, unilateral patients had an average 32% decrease in the platelet count, compared with the 64% decrease seen in bilateral patients. Analysis of fibrin split products revealed a trend toward greater elevation of these degradation products after bilateral procedures. The hematologic changes represented evidence of activation of the coagulation and fibrinolytic systems. These changes tended to be more pronounced in patients treated with bilateral procedures, and were manifested in postoperative elevation of fibrinogen and fibrin degradation products, with reductions in the platelet count. The degree of relative thrombocytopenia raises concerns about careful evaluation of candidates for bilateral arthroplasty, especially when determining the preoperative platelet count. PMID- 8425330 TI - Patellar tilt and subluxation in total knee arthroplasty. Relationship to pain, fixation, and design. AB - Two hundred thirty-four primary total knee prostheses were evaluated with a 45 degrees merchant view to examine patellar position and fixation. Despite a standardized technique for obtaining roentgenographs, patellar position varied with leg position. Overall, 54.7% tracked centrally, 31.2% tilted, and 14.5% displaced. The incidence of these findings was the same whether the patella was domed or congruent. Neither pain scores nor fixation were affected by position. Postoperative tilt and displacement were more common in patellae that were tilted preoperatively. The incidence of postoperative tilt or displacement was not significantly different in knees in which a lateral release was performed. Patellar tilt in some patients seems inevitable despite careful technique because intraoperative tests are static and postoperative function is dynamic. This does not bode well for wear in a metal-backed patella. Tilt caused increased loading at the periphery of the component where most metal-backed prostheses have thin polyethylene. PMID- 8425331 TI - Isolated patellar component revision of total knee arthroplasty. AB - Forty-two knees in 41 patients required isolated patellar component revision of a total knee arthroplasty (TKA). Revision was performed for loosening in 14 knees, wear to metal backing in 13, fracture of the fixation peg in seven, dissociation of polyethylene from metal backing in two, anterior knee pain in two, patellar instability in two, and component malposition with fat pad proliferation in two. Thirty-six knees were evaluated two to eight years after patellar revision. The Hospital for Special Surgery knee scores improved from an average of 71 preoperatively to 81 postoperatively. Eighteen knees were excellent; 12, good; four, fair; and two, poor. Complications associated with patellar revision included late patellar fractures in five knees, patellar instability in three, peroneal nerve palsies in two, patellar polyethylene wear to metal backing in two, infection in one, and extensor lag in one. This seemingly straight-forward procedure is associated with a high complication rate. PMID- 8425332 TI - Total knee arthroplasty without patellar resurfacing. Clinical outcomes and long term follow-up evaluation. AB - Total knee arthroplasty (TKA) without resurfacing of the patella by a single surgeon was reviewed retrospectively in 125 patients: 66 patients (79 knees) were included in the final study group, 32 patients were deceased, 13 were without current address and not able to be contacted, and 14 patients were unable to complete a questionnaire because of severe illness or language barrier. All patients had a diagnosis of osteoarthrosis. The decision to leave the patella unresurfaced was based on the presence of satisfactory patellar articular cartilage, absence of eburnated bone, congruent patellofemoral tracking, normal anatomic patellar shape, and no evidence of crystalline disease or an inflammatory synovitis. A lateral retinacular release was performed in 13 knees (16.5%) to facilitate congruent patellar tracking. The follow-up period averaged 7.5 years (range, 2.4-15.5 years). There were no component revisions and no reoperations. The mean Knee Society score improved from 23.2 to 89.9 postoperatively and the function score improved from 58.4 to 92.0 postoperatively. Mild anterior knee pain was reported in 19%. Satisfaction with the surgical result was expressed in 89.5% of patients; 4% were somewhat satisfied; 2.6% were somewhat dissatisfied; and 4% were neutral. Of the patients who had mixed bilateral TKA (one TKA with unresurfaced patella, the other resurfaced), 46% rated both as equal; 46% preferred the TKA with the resurfaced patella; and 7.7% preferred the unresurfaced patella. In patients meeting the selection criteria outlined above, TKA without resurfacing of the patella provided satisfactory long-term results and a high degree of patient satisfaction with an absence of mechanical complications and no reoperations at the average 7.5-year follow-up evaluation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425333 TI - The effect of femoral component position on the kinematics of total knee arthroplasty. AB - In a laboratory study using seven fresh-frozen anatomic specimen knees, the effect of total knee arthroplasty on the three-dimensional kinematics of the patella, femur, and tibia were measured. Experiments were performed in the intact knee, after division of the anterior cruciate ligament (ACL), after total knee arthroplasty, and after 10 degrees internal rotation, 10 degrees external rotation, 5-mm medial shift, and 5-mm lateral shift of the femoral component on the femur. The presence of a high lateral ridge on the anterior surface of the femoral component effectively prevented patellar subluxation or dislocation, but displaced and tilted the patella medially. Internal rotation or medial displacement of the femoral component exaggerated this medial patellar displacement and shift. External rotation of the femoral component corrected it, except at flexion angles greater than 100 degrees, where the femur was shifted medially on the tibia and externally rotated 15 degrees. This combination produced a net 10-mm medial displacement of the patella relative to the tibia at 120 degrees knee flexion. Lateral placement of the femoral component compensated for the effect of the high lateral ridge and allowed more normal patellar tracking while allowing tibiofemoral motions similar to those seen after sectioning of the ACL. The kinematics of the patellofemoral and tibiofemoral joints were not reproduced with a total knee prosthesis that sacrifices the ACL. When using a prosthesis with a high lateral ridge, lateral placement of a femoral component prevented patellar dislocation and allowed patellar tracking patterns similar to those seen in the intact knee without further altering tibiofemoral motions. PMID- 8425334 TI - The natural history of unicompartmental arthroplasty. An eight-year follow-up study with survivorship analysis. AB - From 1983 to 1987, 82 unicondylar arthroplasties were performed for primary unicompartmental osteoarthrosis. Seven knees had lateral compartment arthroplasties and 75 had medial unicompartmental knee arthroplasties (UKA). At a minimum follow-up period of four years, eight patients had been revised, one patient was scheduled for revision, and one patient had died with a failed UKA, for a total failure rate of 12%. Reasons for failure were progression of tricompartmental arthritis in two patients (2.4%), polyethylene wear in two (2.3%), component failure in two (2.3%), component loosening in three (3.5%), and technical error in one (1.6%). Furthermore, 14 (17%) patients showed roentgenographic evidence of impending failure and an additional 12 (15%) patients showed a gradual decline in Hospital for Special Surgery knee scores. Unicondylar knee arthroplasty does not appear to provide as reliable pain relief as total knee arthroplasty. Progression to tricompartmental arthritis, polyethylene wear, and component loosening were seen to limit the usefulness of this arthroplasty. Unlike total knee arthroplasty, factors affecting the success or failure of UKA cannot be predictably controlled by the surgeon. PMID- 8425335 TI - Unicondylar knee arthroplasty. An evaluation of selection criteria. AB - In a prospective evaluation of 165 consecutive patients (228 knees), intraoperative evaluation of the knees was performed at the time of total knee arthroplasty. Each of the three knee compartments was independently graded for arthritic changes depending on the extent of articular degeneration visualized. Patients were believed to be suitable candidates for unicondylar knee arthroplasty (UKA) if they fulfilled the Kozinn and Scott criteria. Specific attention was given to patient age, weight, preoperative range of motion, angular deformity, as well as the extent of intraoperative cartilage erosions seen. Thirty-five knees (15%) were candidates for UKA based solely on inspection of the articular surfaces at the time of surgery. Further analysis revealed that of these 35 knees, 22 failed to meet the other selection criteria. Thus, of the original 228 knees, only 13 knees (6%) fulfilled all of the stringent selection requirements and were considered suitable candidates for UKA. With proper patient selection, the number of UKAs performed could become relatively small. PMID- 8425336 TI - Unicompartment arthroplasty for osteoarthrosis of the knee. AB - Sixty-eight consecutive unicompartment knee arthroplasties were reviewed. Seven patients were lost to follow-up evaluation, and the review group consisted of 55 patients with 61 medial compartment replacements. Follow-up periods averaged 51 months, with a range of 24 to 70 months. All of the knees had a Robert Brigham unicompartment arthroplasty. All patients were evaluated using The Knee Society Clinical Rating System. Overall results revealed that 70% (42 patients) rated excellent, 10% (seven patients) rated good, 10% (six patients) rated fair, and 10% (six patients) were failures. Preoperative scores in the pain, range of motion, and stability sections averaged 54 points of a maximum of 100. Postoperatively, the scores averaged 87 points. Postoperative roentgenographs were evaluated using The Knee Society Roentgenographic Evaluation and Scoring System. Roentgenographs revealed at least one radiolucent line in 16 femoral components and 15 tibial components, and nine in both. Four of the six failures in the series had lucent lines in both the femur and tibia. This procedure is recommended in active patients between the ages of 55 to 65 years with single compartment disease. Also, a unicompartment arthroplasty is recommended in more sedentary patients 65 years and older with single compartment disease. PMID- 8425337 TI - Methylmethacrylate monomer and fat content in shed blood after total joint arthroplasty. AB - Twenty-five patients were prospectively evaluated to quantify levels of methylmethacrylate monomer and fat in systemic blood and in shed blood after total joint arthroplasty. Levels of methylmethacrylate monomer in systemic blood were measured at intervals after insertion of the prosthesis. Levels of methylmethacrylate monomer in shed blood were measured at intervals after insertion of the drain. Levels of fat in systemic blood were measured preoperatively and 30 minutes after insertion of the prosthesis. Levels of fat in shed blood were measured 60 minutes after insertion of the drain. No significant fat or methylmethacrylate monomer was noted in systemic blood. Levels of methylmethacrylate monomer in shed blood were highest five minutes after insertion of the drain. Levels of methylmethacrylate monomer in shed blood collected from the hip were significantly lower than levels in shed blood from the knee. Levels of shed blood from the hip and knee were undetectable six hours after insertion of the drain. Shed blood from the hips and knees contained fat particles of three diameters: fat particles less than 9 microns, 9-40 microns, and greater than 40 microns. The diameter of most of the fat particles in the shed blood was less than 9 microns. Fat particles less than 40 microns in diameter will not be removed by microaggregate screen filters 40 microns in diameter. PMID- 8425338 TI - Unicompartmental knee arthroplasty. A multicenter investigation with long-term follow-up evaluation. AB - Nonmental-backed, cemented, unicompartmental knee arthroplasty has a survivorship rate in this multicenter investigation at ten years of 91.4% (+/- 2.8). High levels of patient weight were associated with increased need for revision arthroplasty. Overall, men had a lower revision rate (2.4%) compared with women (3.9%). Valgus postoperative alignment was minimally associated with progression of disease as a cause for revision. No difference in revision rates between medial and lateral compartmental arthroplasty was noted. The theoretical clinical benefits of the use of metal-backed tibial components will need to be reevaluated in light of these findings. PMID- 8425339 TI - Cementless revision total knee arthroplasty. AB - This is a report of the outcome of cementless revision total knee arthroplasty (TKA), and describes the technique for bone grafting of massive femoral and tibial defects. The technique relies on rigid fixation of the implants using the remaining shell of distal femur and proximal tibia and the diaphysis of each bone for support of the implant. Fifty-six cementless revision TKAs in 56 patients were performed using cementless technique, long-stemmed components, and morselized allograft between April 1985 and April 1989. All knees had major loss of femoral and tibial bone stock. Fixation of the stemmed implants required firm seating on the rim of the femur and tibia. The stem engaged the isthmus of the bone to provide toggle control, and the screws fixed the implant firmly to bone. Two years after surgery, 30 (54%) of the patients had no pain, 17 (30%) had mild pain, five (9%) had moderate pain, and four (7%) had severe pain. All 56 knees had increasing radiodensity in the grafted areas one and two years postoperatively as compared with the one-month roentgenogram. All implants except two achieved stable fixation to bone. Alignment, stability, and comfort of the knee were all improved with cementless revision technique. The overall results of this study are encouraging. Bone stock was reliably reconstructed, and fixation during the short term appears to be durable. PMID- 8425340 TI - Reoperation after condylar revision total knee arthroplasty. AB - Reasons for reoperation after knee revision surgery are implant loosening, sepsis, extensor mechanism problems, fractures of bone or prosthetic components, wear debris, and limited range of motion. The purpose of this study was to review the complications requiring reoperation in a large number of condylar revision total knee arthroplasties to determine the incidence and outcome after treatment. Six hundred fifty-five condylar revision total knee arthroplasties performed during a ten-year period were retrospectively reviewed. Forty-six knees without a history of arthroplasty infection required a total of 60 reoperations after the revision surgery. A reoperation was performed for extensor mechanism or patellar problems in 19 knees (41%), component loosening in ten knees (22%), deep infection in nine knees (20%), wound problems in nine knees (20%), tibiofemoral instability in eight knees (17%), limited range of motion in four knees (8%), and particulate debris synovitis in one knee (2%). All patients were observed for an average of 7.5 years. Twenty-four knees (52%) were considered clinical failures because of pain, limited motion, instability, and sepsis. Awareness of these failure modes may help to prevent complications by strict attention at the time of revision surgery to protection of the patellar tendon attachment and collateral ligaments, balancing of the extensor mechanism, preservation of the patellar blood supply, proper component position and sizing, restoration of the mechanical axis, and use of more constrained implant designs. PMID- 8425341 TI - Percutaneous diskectomy for lumbar disk herniation. A preliminary report. AB - Percutaneous diskectomy is a new method for reducing lumbar disk herniation. This procedure is simple, safe, and only semiinvasive since it causes no direct damage to the dura or nerve roots. This technique employs a nucleotome (blunt-tipped, suction-cutting probe) with a rotating electric shaver (3 mm in diameter) that was specially developed for this procedure. It was applied to 117 patients with lumbar disk herniation whose sciatica had not been relieved by conservative treatments. The results of the technique were considered effective in 94 patients (80.3%). The improvements were even more marked in patients with protrusion or prolapse type herniation. Reviewed with postoperative examinations using magnetic resonance imaging (MRI), excision of the herniated disk, resulting in decompression of the nerve root, may be correlated with the relief of symptoms. PMID- 8425342 TI - Vascular ultrasonography for deep venous thrombosis after total knee arthroplasty. AB - Screening vascular ultrasonography was performed postoperatively on 164 consecutive patients being treated with total knee arthroplasty (203 total knee prostheses). This consisted of examination of the femoral and popliteal veins of the operative extremity with color flow and duplex ultrasonography one week postoperatively. All patients received deep venous thrombosis (DVT) prophylaxis with sequential compressive pneumatic stockings, low-dose warfarin, continuous passive motion, and early mobilization. All patients were observed prospectively for thromboembolic sequelae for a minimum of six months postoperatively. The screening study was significantly limited in six (3%) of the 203 total knee prostheses. The overall incidence of sonographically detected proximal DVT or symptomatic calf vein thrombosis was 3% (6/203), and the incidence of symptomatic pulmonary embolism was 2% (4/203). Four of the ten thromboembolic complications occurred after hospital discharge. Results of this study suggest that duplex ultrasonography can be a useful screening method for identification of venous thrombosis after TKA. Patients with asymptomatic proximal DVT can be identified and appropriately treated before development of serious thromboembolic complications. Routine screening for DVT after TKA can avoid the considerable expense, inconvenience, and potential risk of complications associated with prolonged postoperative prophylactic anticoagulation. The combination of sequential compressive stockings, early mobilization, and low-dose warfarin appears to be a safe and effective prophylactic regimen against venous thromboembolic disease in these high-risk patients. PMID- 8425343 TI - A computed tomographic analysis of changes in the spinal canal after anterior lumbar interbody fusion. AB - Thirteen patients with spondylolisthesis (six isthmic type and seven degenerative type) and ten patients with intervertebral disk herniation were treated by anterior lumbar interbody fusion. Preoperative and postoperative computed tomography (CT) scans were performed for each patient, and changes in anteroposterior (AP) diameter and lateral diameter of the dural sac, the area of the dural sac, and the amount of disk bulging were measured. The periodic tomogram was done in all patients postoperatively, and one- and two-year fusion rates were calculated. The calculations were compared with the early and late clinical results. The early clinical results after operation were excellent in 26.1%, good in 56.5%, and fair in 17.4%. The late clinical results were similar to early results. The early clinical results correlated with the changes in the spinal canal, such as an increase in AP diameter of the dural sac and a decrease in amount of disk bulging after anterior interbody fusion. There was statistical correlation between the early clinical results and the change in AP diameter of the dural sac. But the late clinical results were influenced by multiple factors including solid bony fusion. PMID- 8425344 TI - Computer graphic analysis of endosteal expansion after total hip arthroplasty. AB - Femoral endosteal expansion is known to occur naturally with aging and may contribute to femoral component loosening after cemented total hip arthroplasty (THA). Previous roentgenographic models used to quantify femoral expansion have used a limited number of measurements at arbitrary axial sites in one roentgenographic view. A computer graphic model is introduced, which uses standard anteroposterior (AP) and lateral roentgenograms to reconstruct a three dimensional model of the femur between the lesser trochanter and the distal end of a cemented prosthesis. The model was used retrospectively to calculate endosteal and periosteal surface area and volume at the time of surgery and at follow-up examination for 22 patients who had been treated with hip arthroplasty (average, 9.3-year follow-up observation). Endosteal surface area and volume were found to increase an average of 3.1 cm2 and 1.7 cm3, respectively. Periosteal dimensions did not change significantly. Overall endosteal expansion occurs in the vicinity of the implanted prosthesis. The extremely low interobserver variability indicates that this model provides a reliable method of following surface area and volume changes around a cemented hip prosthesis. PMID- 8425346 TI - Total knee arthroplasty. PMID- 8425345 TI - Use of pulsed electromagnetic fields in treatment of loosened cemented hip prostheses. A double-blind trial. AB - A double-blind trial of pulsed electromagnetic fields (PEMFs) for loosened cemented hip prostheses was conducted at two centers. Of the 40 patients who enrolled, 37 met entry criteria and were available for analysis. All patients completed six months of treatment (either active or control units). Success was determined clinically by a Harris hip score greater than or equal to 80 points (or an increase of ten points if initially greater than or equal to 70 points). Ten of the 19 active units were successes (53%), whereas two of the 18 controls (11%) exhibited a placebo effect, a statistically significant and clinically relevant result. A 60% relapse rate among the active successes was seen at 14 months poststimulation, and despite maintenance therapy of one hour per day, the relapse rate increased to 90% at three years. These data suggest that for loosened cemented hip prostheses, use of PEMFs is a treatment option only to delay revision hip surgery. PMID- 8425347 TI - Survivorship analysis of hemiarthroplasties. AB - From 1976 until 1988, 679 hemiarthroplasties were inserted for displaced femoral neck fractures. Two hundred two were of the Austin Moore design (one-piece, noncemented), 209 of the Christiansen design (trunion bearing, cemented), and 268 of the Hastings design (bipolar, cemented). Five years after the hemiarthroplasty, the cumulative proportion of prostheses surviving was 90%, declining to 85% after ten years. A survivorship analysis indicated that the cemented bipolar Hastings prosthesis and old age (75 years or older) were two variables associated with significantly less failures. The survivorship analysis did not indicate that the group differences were influenced by improvements in surgical techniques over time. Gender and social setting had no influence on the survival of prostheses. Cemented bipolar hemiarthroplasty should be considered as an alternative in the internal fixation of displaced femoral neck fractures in the elderly. PMID- 8425348 TI - Acute arthritis of the hip in a child infected with the Lyme spirochete. AB - Acute Lyme arthritis may mimic acute pyogenic arthritis. Although the arthritis associated with infection with the spirochete, Borrelia burgdorferi, is more commonly seen in the chronic stage (Stage III) of the disease, occasionally it may present as the initial clinical manifestation. A five-year-old girl with acute arthritis of the hip is reported to discuss classification and management of arthritis associated with Lyme disease. PMID- 8425349 TI - Roentgenographic diagnoses of ruptured Achilles tendons. AB - In a prospective study, the roentgenographic features of the Achilles tendon were investigated in 60 patients with acute total rupture of the Achilles tendon. Sixty ankles without actual injuries, 60 patients with ankle fractures, and 60 patients with ankle sprains were selected as reference groups. At primary examination, 11 patients (18%) with Achilles tendon rupture were clinically estimated to have minor lesions in the Achilles tendon; all had a correct roentgenographic diagnosis. Kager's triangle was positive for rupture of the Achilles tendon in all 60 patients with operative verification of the rupture, seven (12%) had diminished Toygar's angle, and 29 (48%) had positive Arner's sign. Forty-seven patients with ruptured Achilles tendon had a thickness of the tendon compared with the opposite Achilles tendon. To lower the frequency of overlooked Achilles tendon rupture, any doubt as to whether a rupture of the Achilles tendon is present should result in a lateral roentgenographic examination of the ankle. Furthermore, the authors conclude that the presence or absence of Kager's triangle should be determined roentgenographically. If Kager's triangle is not identified on plain lateral roentgenographs of ankle, special soft tissue roentgenographs may be useful. PMID- 8425350 TI - Irreducible talar dislocation with entrapment by the tibialis posterior and the flexor digitorum longus tendons. A case report. AB - Complete open-talar dislocation irreducible by virtue of entrapment by the tibialis posterior and flexor digitorum longus tendons occurred in a 41-year-old woman. Two-year follow-up examination revealed no evidence of avascular necrosis. A cursory review of the literature suggests that recovery of a complete range of motion is unusual. PMID- 8425351 TI - The role of surgical therapy in patients with nonmetastatic Ewing's sarcoma of the limbs. AB - Of 131 patients with primary Ewing's sarcoma (ES) with extremity locations, treated between April 1972 and May 1987, 13 had amputation; 56 had a local resection with 31 (55%) requiring radiation therapy (RT) as an adjuvant to local therapy. Sixty-two of 131 patients (47%) had RT only. Local recurrences were reported in 5% of patients treated with surgery alone, 1% of the group treated with surgery and radiation therapy, 24% of patients treated with radiation therapy alone (46% for cesium-radiation therapy, and 12% with cobalt-radiation therapy). PMID- 8425352 TI - A system for the functional evaluation of reconstructive procedures after surgical treatment of tumors of the musculoskeletal system. AB - The need for a standardized system of end result reporting of various surgical alternatives after limb salvaging and ablative procedures for musculoskeletal tumors was clearly recognized during the first International Symposium on Limb Salvage (ISOLS) in 1981. During the ensuing four biannual symposia, there has been an ongoing developmental experience with a system extensively field tested in 1989 by the Musculoskeletal Tumor Society (MSTS). This system of functional evaluation has been adopted by the MSTS and ISOLS for their joint studies and program presentation. In brief, the system assigns numerical values (0-5) for each of six categories: pain, and function and emotional acceptance in upper and lower extremities; supports, and walking and gait in the lower extremity; and hand positioning, and dexterity and lifting ability in the upper extremity. Demographic information and a patient satisfaction component is included. A numerical score and percent rating is calculated to allow for comparison of results. The system has been field tested in 220 patients with low (+/-) interobserver variability. It was well accepted by the participants, and its usage is recommended by the MSTS to facilitate valid comparative end result studies of musculoskeletal tumor reconstructions. PMID- 8425353 TI - Functional outcome of below-knee amputation in peripheral vascular insufficiency. A multicenter review. AB - A retrospective functional outcome study was performed on 299 patients treated with below-knee amputations for peripheral vascular disease during a three-year period in six Veterans Administration special amputee centers. Sixty-one percent of the patients were diabetic, and 25% were previously unilateral amputees. Surgery was performed with a long posterior flap in 92.3%, sagittal flaps in 20%, and guillotine open technique in 5.7%. Wound management was accomplished with rigid plaster dressings in 75.3%, pneumatic compression dressing in 14%, and soft dressings in 10.7%. At follow-up evaluation one to two years later, 36.1% had died. Thirteen percent suffered wound complications, with 6.7% requiring revision to a more proximal amputation level. Eighty-seven percent of community ambulators maintained their ambulatory status. A comparable percentage maintained their ability to walk with a prosthesis. PMID- 8425354 TI - A survey of patients with Ehlers-Danlos syndrome. AB - Ehlers-Danlos syndrome (EDS) is an inherited disorder of collagen biosynthesis. Nine types of the disorder are now recognized. A survey of 151 EDS patients showed a high percentage of the patients were classified as having Types I through IV. Bracing and fusion appear to be the most commonly used methods of orthopedic care. Surgical complications were common. The orthopedist was often the first practitioner to diagnose and treat the patient with EDS. A fundamental understanding of this complex disorder and its varied manifestations is essential to prevent major life-threatening complications. PMID- 8425355 TI - Occlusion of arterial flow in the extremities at subsystolic pressures through the use of wide tourniquet cuffs. AB - Tourniquet-induced peripheral neuropathy is at least partially attributable to excessive forces applied to the nerves beneath cuffs inflated to high pressures. Lowering the inflation pressure to the minimum necessary to obtain an effective arrest of blood flow distal to the tourniquet cuff should increase the safety of these systems. Tourniquet cuffs with widths varying from 4.5 cm to 80 cm were applied to the upper and lower extremities of 34 healthy, normotensive volunteers. Occlusion pressure for the arterial system under study was estimated by determining that level of cuff inflation at which the distal pulse became detectable by ultrasonic flowmetry. The occlusion pressure was inversely proportional to the ratio of tourniquet cuff width to limb circumference and was in the subsystolic range at a cuff width to limb circumference ratio above 0.5. Wide tourniquet cuffs can achieve an effective arrest of the regional arterial circulation at subsystolic pressures of inflation. Wide cuffs may reduce the risk of tourniquet-induced injury to underlying soft tissues by lowering the inflation pressure required to secure a bloodless field. PMID- 8425356 TI - Transplant of bone marrow and muscle-derived connective tissue cultures in diffusion chambers for bioassay of bone morphogenetic protein. AB - To promote a high concentration gradient, syngeneic bone marrow and muscle connective tissue cell cultures and subcultures were exposed to bone morphogenetic protein and associated noncollagenous bone matrix proteins (BMP/NCP) in vitro, transferred to diffusion chambers, and then transplanted into the anterior abdominal wall of isogeneic rats. Both muscle- and marrow-derived cells differentiated into cartilage and chondroosteoid on the inside of diffusion chambers. New bone developed on the vascularized outside surfaces in juxtaposition to avascular tissue on the inside. Levels of glycosaminoglycans (GAGs) and DNA synthesis in tissues inside the chambers were sharply elevated. These data, correlated with histologic observations, demonstrate that the BMP/NCP recruits mesenchymal-type cells for a skeletal tissue pathway of development. Muscle-derived cells, as distinguished from marrow-derived cells, originate in a nonosseous environment but differentiate into osteoprogenitor (osteogenic stem) cells and display levels of DNA synthesis almost as high as marrow stroma-derived cells. The rise in DNA synthesis was maximal in the interval from two to three weeks after transplantation. Whether the bone marrow stroma mesenchymal cell target for BMP is only the osteogenic stem cell population or also includes colony-forming units fibroblasts (CFU-F) remains to be investigated by experiments on established cell lines. PMID- 8425357 TI - Incidence of pulmonary embolism after total knee arthroplasty with low-dose coumadin prophylaxis. AB - The incidence of pulmonary embolism (PE) in osteoarthritic patients prophylaxed with low-dose coumadin after cemented total knee arthroplasty (TKA) was investigated prospectively. Each patient had a preoperative perfusion scan and a ventilation-perfusion scan on the seventh postoperative day. Pulmonary embolism was diagnosed by a high probability ventilation-perfusion scan or positive arteriogram. Patients with a moderate probability scan had an arteriogram to rule out PE. Pulmonary embolus was identified in 48 (5.6%) of 852 TKAs in 755 patients. Of these, six (0.7%) were symptomatic, and no fatal PE was identified. Age, gender, and weight did not show statistical differences comparing the PE and non-PE groups, nor did the incidences of previous PE, contralateral phlebitis, malignancy, and diabetes. A history of ipsilateral phlebitis increased the risk of PE from 5.2% to 13%, and a history of cardiac disease decreased the risk from 7.8% to 4.2%. Type of anesthesia, blood loss, tourniquet time, and prosthesis type were not significant factors. With the exception of previous contralateral phlebitis, traditional risk factors for PE were not found to increase risk of PE with low-dose coumadin prophylaxis. Spinal anesthesia that has been shown to be protective in total hip surgery was not a significant factor in this study. PMID- 8425358 TI - Staging of patellar tendon autograft healing after posterior cruciate ligament reconstruction. A biomechanical and histological study in a sheep model. AB - The prerequisite for adequate rehabilitation after cruciate ligament reconstruction with a patellar tendon autograft is a thorough knowledge of the biologic healing processes. It is of interest to determine whether distinct phases similar to those in wound healing can be differentiated. It is also important to assess the magnitude of the biomechanical loading capacity and the duration of the healing processes. One posterior cruciate ligament in each of 48 skeletally mature sheep was replaced with a free patellar tendon autograft. Immediate rehabilitation without immobilization followed. Four phases of healing were demonstrated using the histologic condition of the autograft as a guide. The biomechanical data were correlated with the morphologic data. During the necrotic phase, a maximum of necrotic tissue was seen two weeks postoperatively. The strength of the graft is initially limited by the surgical fixation strength; however, later the intraarticular portion becomes the strength-limiting factor. During the revitalization phase, which is characterized by revascularization and proliferation of fibroblasts, and during the following phase, collagen formation, an increase in maximum stress is seen while the elastic modulus remains constant. Only in the remodeling phase is an increase in elastic modulus seen, owing to longitudinal alignment of the collagen bundles. One year after implantation, the autograft achieves approximately 50% of the material properties of the control. Even after two years, the autograft reaches only a maximum stress of 60% and an elastic modulus of 70% of the control. Ligamentization of the autograft could not be demonstrated in this study, but degeneration was seen in the core region of the graft during late remodeling. PMID- 8425359 TI - Penetration and shear strength of cement-bone interfaces in vivo. AB - Using sham replacement of the proximal femur in adult mongrel dogs, shear strength at the interface between polymethylmethacrylate bone cement and cancellous bone has been found to be linearly dependent on the depth of penetration of the cement into the bone. Shear strength at the interface was increased by 82% and penetration by 74% when distal bone plugging, pressure lavage, and pressurized insertion of cement were employed. Use of a lower viscosity cement gave a further 18% increase in penetration and shear strength. There was no film of blood at the cement-bone interface with pressurized insertion of Simplex P and Palacos R cements. PMID- 8425360 TI - The effect of wound environment on the incidence of acute osteomyelitis. AB - A model was developed to identify and compare the local wound factors that induce acute osteomyelitis in a prospective, controlled investigation. When compared with wounds containing either virulent bacteria or dead bone, statistical analysis disclosed a significant increase in the incidence of osteomyelitis when virulent bacteria and dead bone were combined. The incidence of osteomyelitis in wounds containing an inoculated, hematoma-filled dead space was significantly less when compared with wounds containing dead bone and virulent bacteria. The incidence of osteomyelitis is significantly less when a nonvirulent strain of bacteria is substituted for a virulent strain. Although rigid internal fixation increased the incidence of osteomyelitis to 100% and long-term antibiotic therapy decreased the incidence, these changes were not statistically significant. These data allow the authors to predict the relative risk of osteomyelitis when these wound factors are present. The prevention of osteomyelitis depends on the clinical identification and modification of these local wound factors. PMID- 8425361 TI - The influence of basal cartilage calcification on dynamic juxtaarticular stress transmission. AB - Dynamic finite element analysis is used to elucidate the manner in which juxtaarticular stress distributions depend on the transitional mechanical properties of the calcified cartilage layer. A finite element model is used to study how these stresses change in response to the thinning of the articular cartilage layer associated with tidemark advancement. The finite element results indicate that shear stress levels within the deepest layer of articular cartilage are increased when tidemark advancement with concomitant cartilage-thinning and calcified cartilage/subchondral plate thickening is modeled. The cartilage thinning associated with tidemark advancement, observed both clinically and experimentally, may be an explanation for what has previously been considered wear-related thinning. PMID- 8425362 TI - Acetabular dysplasia in the adolescent and young adult. PMID- 8425363 TI - Minor anatomic abnormalities of the hip joint persisting from childhood and their possible relationship to idiopathic osteoarthrosis. PMID- 8425364 TI - Intramedullary versus extramedullary femoral alignment systems in total knee replacement. AB - The accuracy of intramedullary (IM) versus extramedullary (EM) distal femoral alignment systems was compared in 200 consecutive total knee replacements (TKRs) on standing full-length lower-extremity roentgenographs. Intramedullary femoral alignment systems were used in 125 TKRs and an EM system was used in 75 TKRs. All tibial cuts were made using an EM tibial cutting guide. Roentgenographic measurements of (1) femoral-tibial angle, (2) distal-femoral resection angle, (3) proximal-tibial resection angle, (4) joint line orientation, (5) physiologic femoral valgus, and (6) distance of the lower-extremity mechanical axis from the center of the knee were made. No significant intragroup differences were seen in the average values obtained for each of the six roentgenographic measurements. However, the percentage of distal-femoral resections outside the accepted normal range (94 degrees-100 degrees) was higher in the EM group (28%) versus the IM group (14.4%) at p = 0.019. Likewise, the percentage of joint line orientations outside the desired normal range was higher in the EM groups when compared with the IM group (21.3% versus 11.2%, p = 0.052). A disturbing number of proximal tibial resection angles were inaccurate in both groups. Improper tibial cuts were seen in 20.6% of Group 1 and in 24% of Group 2 when an EM tibial cutting guide was used. A range of distal-femoral cuts exists when using either an IM or an EM femoral alignment guide. This study demonstrated a statistically significant improvement in distal-femoral resection accuracy when using an IM femoral alignment system. Methods of improving proximal-tibial bone resections are needed.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425365 TI - Lesions of the tibial tubercle occurring during adolescence. 1903. PMID- 8425366 TI - Determining the rotational alignment of the femoral component in total knee arthroplasty using the epicondylar axis. AB - The posterior condylar surfaces of the femur are routinely used as the reference for the rotational orientation of the femoral component during most primary total knee arthroplasties. The purpose of this investigation was to identify a clearly discernible, reproducible secondary anatomic axis useful for determining the rotational orientation of the femoral component when the posterior condylar surfaces cannot be used. Seventy-five embalmed anatomic specimen femurs were studied. A surgical epicondylar axis was defined as the line connecting the lateral epicondylar prominence and the medial sulcus of the medial epicondyle. The posterior condylar angle was measured as the angle between the posterior condylar surfaces and the surgical epicondylar axis. Measurement of the posterior condylar angle referenced from the surgical epicondylar axis yielded a mean posterior condylar angle of 3.5 degrees (+/- 1.2 degrees) of internal rotation for males and a mean posterior condylar angle of 0.3 degree (+/- 1.2 degrees) of internal rotation for females. Thus, rotational alignment of the femoral component can be accurately estimated using the posterior condylar angle. The posterior condylar angle, referenced from the surgical epicondylar axis, provides a visual rotational alignment check during primary arthroplasty and may improve alignment of the femoral component at revision. PMID- 8425367 TI - V-Y quadricepsplasty in total knee arthroplasty. AB - The results in 14 consecutive patients (16 knees) who have had total knee arthroplasty (TKA) with a V-Y quadricepsplasty were reviewed. There were eight men and six women with an average age of 66.2 years. Nine patients (ten knees) had revision arthroplasty and five patients (six knees) had primary arthroplasty. Nine of the 14 patients were examined and evaluated in the biomechanics laboratory using the modified Cybex-II system at a mean of three years from surgery. Using the knee rating score of the Hospital for Special Surgery, there were two excellent, ten good, two fair, and two poor results. The average active range of motion was 4 degrees to 85 degrees. Biomechanical testing revealed statistically significant weakness in extension compared with the contralateral side only at test speeds of 120, 180, and 240 degrees per second in those patients who had contralateral normal knees. The affected side at other test speeds were weaker but did not reach statistically significant weakness when compared with the contralateral knee. Comparing V-Y quadricepsplasty to normal medial parapatellar TKA patients, the extensor mechanism was weaker in the V-Y group but not to a significant degree. If one requires a V-Y quadricepsplasty during TKA, one can expect near normal active extension and moderate weakness in extension. Knee scores reflect the difficult reconstructive challenges that are inherent in patients who require a V-Y quadricepsplasty. PMID- 8425368 TI - Avoiding neurologic and vascular injuries with screw fixation of the tibial component in total knee arthroplasty. AB - This investigation determined the position of neurologic and vascular structures at risk during screw fixation of an uncemented tibial component in total knee arthroplasty relative to the cut surface of the tibial plateau. Sixteen anatomic specimens of the lower extremities were studied by systematically placing screws about the tibial plateau followed by computed tomographic scanning with three dimensional reconstruction and anatomic dissection. The structures at risk during screw preparation and placement were the tibial nerve, popliteal artery, popliteal vein, anterior tibial artery, and common peroneal nerve with its superficial and deep branches. The application of a clock system to the cut surface of the tibial plateau with the six o'clock position defined as directly anterior and the three o'clock position defined as lateral identified the entire region between the 11 o'clock and three o'clock positions as a location where potential injury to vital structures can occur during screw placement. The risk of injury to these structures may be minimized by directing screws radially inward to avoid violation of the tibial cortex. Any penetration of the cortex between the 11 o'clock and three o'clock positions may have potentially devastating consequences because of the close proximity of neurologic and vascular structures. PMID- 8425369 TI - Gastrocnemius muscle flap coverage of exposed or infected knee prostheses. AB - A retrospective analysis was performed of 12 patients who required soft-tissue coverage of an exposed or infected total knee prosthesis between 1982 and 1989. All knees had skin closure with medial gastrocnemius muscle flaps. At a mean follow-up period of 41 months, all patients who were treated for infected prostheses with removal of the implant, intravenous antibiotics, and muscle flap closure had an excellent clinical result with complete skin coverage and no infection. Five of the six patients went on to successful reimplantation. Of the patients with an exposed prosthesis, five of six had an excellent outcome with retention of the prosthesis. Thus, 11 of 12 patients (92%) who had medial gastrocnemius flap coverage of an exposed or infected knee prosthesis had an excellent outcome, with ten of 12 patients (82%) retaining their prostheses or having a successful reimplantation. No medial gastrocnemius flap failed after standard primary or revision total knee arthroplasty. Gastrocnemius muscle flaps provide excellent soft-tissue coverage of exposed or infected knee prostheses and facilitate surgical care of this difficult problem. PMID- 8425370 TI - Fracture of the femoral component in cementless total knee arthroplasty. AB - Records of sales revealed that 6172 Ortholoc II cementless total knee femoral components with double bead layers were sold between January 1986 and December 1987. Sixteen thousand two hundred thirty Ortholoc II femoral implants with single bead layers were sold between December 1986 and January 1990. Fractures of the femoral components were reported in 32 knees; all but one occurred in femoral components with porous surfaces consisting of two layers of sintered beads. The overall minimum rate of failure for all sizes with double bead layers was 0.42%. The rate varied with the size of the implant and was much greater in smaller implants. The rate of failure was 0.82% in 852 small components, 0.37% in 2416 medium components, 0.29% in 1729 large components, and 0.09% in 1175 extra-large components. The differences in failure rates were statistically significant for all sizes of implants when compared with other sizes. Sixteen thousand two hundred thirty Ortholoc II components with a single layer of beads were sold worldwide between December 1986 and January 1989. One implant (medium size) failed 18 months after surgery. The minimum rate of failure in the single bead layer implants is 0.006%. This failure rate is statistically significantly less than that of the double bead layer implants. All failures occurred at the junction between one of the bevel surfaces and the distal surface of the implant. In all cases, crack propagation was initiated at the inner surface. This suggests that bending stresses had developed and had opened (straightened) the femoral component.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425371 TI - Hybrid total knee arthroplasty. Two- to five-year results using the Miller Galante prosthesis. AB - Forty-one "hybrid" Miller-Galante total knee prostheses having porous-coated femoral and patellar components and a tibial component without a keel, cemented using low-viscosity technique, were implanted and prospectively evaluated for two to five years. The surgical technique was accurate, restoring the mechanical axis of the lower extremity to an average of 1.6 degrees varus. The average postoperative knee score was 90 points with 88% good or excellent results and 88% completely painless. Range of motion improved from a mean 88 degrees to a mean 108 degrees. Nonprogressive, incomplete radiolucent lines were present at the bone prosthesis interface in 27% of patellar, 15% of femoral, and 20% of tibial components. There were six patellar component fractures, four of which have been revised. These clinical and roentgenographic results support the "hybrid" technique for total knee arthroplasty. However, the use of the porous-coated, metal-backed patellar component is not recommended. PMID- 8425372 TI - Cementless total knee arthroplasty in juvenile onset rheumatoid arthritis. AB - Twenty-two total knee arthroplasties with at least one cementless component were performed in 14 patients with juvenile rheumatoid arthritis (JRA) from 1985 to 1989. All 22 femoral components and ten tibial components were implanted cementless. The mean age at operation was 26 years. All 14 patients were available for follow-up evaluation at an average of 3.9 years (range, two to 6.2 years). Using The Knee Society's scoring system, the knee score improved from an average of 18 points (range, 0-47 points) preoperatively to 92 points (range, 58 100 points) at follow-up evaluation. The functional score improved from 28 points (range, 0-55 points) to 76 points (range, 40-100 points). Nonprogressive radiolucencies of less than 1 mm were observed in two knees. One reoperation was performed for failure of a metal-backed patellar component. Knee arthroplasty with cementless components in selected JRA patients can give results comparable with a fully cemented knee at the two- to six-year follow-up evaluation. PMID- 8425373 TI - Long-term results of the total condylar knee arthroplasty. A 15-year survivorship study. AB - This study reports the 15-year survivorship of 112 consecutive Total Condylar knee arthroplasties that have been followed since 1974. Two endpoints were chosen for survivorship: (1) Revision attributable to septic or aseptic loosening or malalignment. (2) Revision or roentgenographic evidence of component loosening. Life table analysis reveals a 94.1% clinical survivorship at 15 years, with an 90.9% survivorship when roentgenographic failures are included. There were five revisions: one for infection, one for instability, and three for tibial loosening. In addition, two tibiae and one patella were considered roentgenographically loose, but were not symptomatic. As of May 1992, 34 patients with 48 knees are known deceased, 15 knees are lost to follow-up evaluation, and 49 knees are available for clinical evaluation. Follow-up data was available on 62 knees for greater than 11 years. Ninety-two percent had good or excellent results, with 1.6% fair and 6.5% poor. Average range of motion was 99 degrees. The average Hospital for Special Surgery knee score was 85. Roentgenographic study revealed lucencies around 72% of tibiae, but only two components were loose. There was a correlation between body weight and the presence of radiolucencies, and patients who weighed more than 80 kg had the lowest survivorship at 15 years: 89.2% clinical survival and 70.6% clinical plus roentgenographic survival. Total Condylar knee arthroplasty has a 94.6% clinical survival at 15 years, with predictably good clinical results. PMID- 8425374 TI - Performance of a fluorescence polarization immunoassay for teicoplanin in serum. AB - Teicoplanin is a new glycopeptide antibiotic that is structurally related to vancomycin, but has six major components and is > 90% protein bound. We have developed a competitive homogeneous fluorescence polarization immunoassay (FPIA) for the quantitative determination of teicoplanin in serum. The teicoplanin FPIA uses six serum-based calibrators over the range 0-100 micrograms/ml, with the lowest teicoplanin concentration at 5 micrograms/ml and a lower limit of detection of < 1.5 micrograms/ml. Samples with concentrations > 100 micrograms/ml can be analyzed after 1:5 dilution with serum. Intra- and interassay precision are < 2% and < 4.5%, respectively, for controls at 7.5, 35, and 75 micrograms/ml on an automated FPIA analyzer. Recovery results for 30 samples prepared from the teicoplanin analytical reference standard over the range 3.4-358 micrograms/ml were linearly correlated with target concentrations; the linear regression equation and correlation coefficient are FPIA = 0.976 x target + 0.120, r = 0.999. Abnormal levels of bilirubin, hemoglobin, albumin, triglycerides, or cholesterol did not significantly affect recovery. Cross-reactivity with vancomycin was < 0.2%, and there was no significant interference from above therapeutic levels of likely concomitant medications. Results on patient samples from the teicoplanin FPIA were highly correlated with both a high-performance liquid chromatography (HPLC) method (FPIA = 1.032 x HPLC + 2.79, r = 0.979, n = 203) and the reference teicoplanin microbiologic assay method (orthogonal regression; FPIA = 1.111 x bioassay + 2.107, r = 0.969, n = 1014). A version of the teicoplanin FPIA for use with a modular clinical research FPIA analyzer showed comparable performance in all respects to the automated assay. PMID- 8425375 TI - Recovery of clinically important microorganisms from the BacT/Alert blood culture system does not require testing for seven days. AB - Recently, we published a comparison of the BacT/Alert blood culture system with the BACTEC 660/730 nonradiometric blood culture system using blood inocula of 5 ml per bottle. By reanalyzing data collected during that study, we found that, for true-positive isolates causing bacteremia or fungemia, 363 (97.6%) of 376 and 341 (97.7%) of 349 isolates were recovered by the end of day 5 of testing, and 364 (97.9%) of 376 and 343 (98.3%) of 349 isolates were recovered by the end of day 6 of testing for aerobic and anaerobic bottles, respectively. Most isolates recovered on days 6 (24 of 27) and 7 (20 of 25) of testing were either contaminants or indeterminate as a cause of sepsis. When used as recommended by the manufacturer, only six (1.3%) of 464 clinically important isolates recovered on test days 6-7 would have gone undetected had testing been limited to 5 days and four (0.9%) of 464 had testing been limited to 6 days. We conclude that BacT/Alert bottles can be tested for as few as 5 days and then discarded with minimal loss of true-positive isolates and maximal reduction of contaminants. PMID- 8425376 TI - Plasmid DNA analysis, biotyping, and antimicrobic susceptibility as subtyping tests for Klebsiella pneumoniae and Klebsiella oxytoca. AB - We compared plasmid DNA analysis, biotyping by Vitek, and disk diffusion antimicrobic susceptibility as subtyping tests of Klebsiella pneumoniae and Klebsiella oxytoca. The 92 tested isolates were from alternate, culture-positive patients over 6 months. No outbreak or cluster of infections was recognized during this interval. Plasmid DNA was detected in 85% of the isolates. Each isolate except one had a reproducible absence of plasmid DNA or a reproducible plasmid DNA profile on repetitive testing. Restriction endonuclease enzyme analysis of plasmid DNA was necessary to distinguish differences among some isolates that had only large plasmids. Isolates with only large plasmids represented 18% of the collection. Of the 78 isolates with plasmid DNA, all but two were considered different from one another by plasmid DNA analysis. Biotyping and antimicrobic susceptibility testing were not highly reproducible. In addition, biotyping did not demonstrate a sufficient variety of patterns among the isolates for subtyping purposes. We conclude that plasmid DNA analysis is very useful as a subtyping test for isolates of K. pneumoniae and K. oxytoca. Neither biotyping nor antimicrobial susceptibility as performed in our laboratory had sufficient discriminatory power and reproducibility for subtyping these organisms. PMID- 8425377 TI - Identification of species-specific, non-cross-reactive proteins of Borrelia burgdorferi. AB - The low specificity of diagnostic tests for Lyme disease is due to the fact that Borrelia burgdorferi possesses many antigenic proteins that are cross-reactive with other spirochetes and bacteria. The low sensitivity is a result of high (> or = 1:100) dilutions used for patient sera during testing to eliminate non specific cross-reactivity. The present study was conducted to identify species specific non-cross-reactive protein(s) of B. burgdorferi that might be used as antigen(s) in serologic tests. Whole-cell sonicates of B. burgdorferi were tested against pooled sera from patients with symptoms, signs, and serologic features diagnostic of Lyme disease (LD), rheumatoid arthritis, infectious mononucleosis, systemic lupus erythematosus, Rocky Mountain spotted fever, secondary syphilis, and from healthy individuals. Different LD pools were also tested against whole cell sonicates of Treponema pallidum, Treponema phagedenis, Leptospira interrogans, and Escherichia coli. Comparison among patterns obtained by each serum pool revealed that IgM antibodies to species-specific 39-, 23-, and 22-kD proteins and IgG antibodies to 34- and 31-kD proteins were present only in the patients with LD and absent from patients with rheumatoid arthritis, infectious mononucleosis, systemic lupus erythematosus, Rocky Mountain spotted fever, secondary syphilis, and healthy individuals pools. These results suggest that 39 , 23-, and 22-kD proteins may be used in an IgM immunoassay for diagnosis of LD. PMID- 8425378 TI - Value of the Hybritech ICON Candida Assay in the diagnosis of invasive candidiasis in high-risk patients. AB - A total of 314 sera from 114 patients at risk for invasive candidiasis were analyzed for the presence of antigenemia using the Hybritech enzyme immunoassay (EIA) for detection of Candida mannan in serum (ICON Candida Assay, Hybritech Inc., San Diego, CA). Fourteen patients (12%) had invasive candidiasis documented by positive blood cultures, deep biopsy culture, and histopathology or autopsy, and five patients had probable invasive candidiasis based on a single positive blood culture and no additional signs of candidiasis. Nine patients had candiduria, 43 patients had mucous membrane colonization, 25 patients were not colonized but received empiric amphotericin B, and 18 patients were not colonized and not treated with amphotericin B. All sera were enzymatically extracted, heat treated, and reacted in a solid-phase sandwich EIA. Test results were read visually and with the ICON reader. The sensitivity and specificity of the mannan EIA in detection of documented invasive candidiasis was 86% and 92%, respectively. The positive predictive value was 60% and the negative predictive value was 98%. Among all patients with invasive candidiasis (documented plus probable), the sensitivity was 68%, the positive predictive value 62%, and the negative predictive value 94%. Specimens were positive within 3 days of the first positive culture in 11 (79%) of 14 patients with documented invasive candidiasis. PMID- 8425379 TI - Tuberculous fasciitis and tenosynovitis. An unusual presentation of miliary tuberculosis. AB - A case of miliary tuberculosis presenting initially as pyogenic fasciitis and tenosynovitis is described. The unusual presenting clinical features suggestive of a noninfectious inflammatory condition resulted in delayed diagnosis. Tuberculous fasciitis and tenosynovitis were diagnosed by the presence of acid fast bacilli in histopathologic sections and confirmed by the growth of Mycobacterium tuberculosis from cultures of tissue specimens. PMID- 8425380 TI - A simple method to prepare Salmonella (A-E) polyvalent antiserum. AB - The preparation of Salmonella O serogroup (A-I) polyvalent antiserum is laborious. In an alternate method, rabbit antiserum raised to cell surface proteins of Salmonella serotype typhi was compared with a commercial Salmonella polyvalent (A-I) antiserum; 51 Salmonella (A-E) and 16 strains of related organisms tested gave comparable agglutination with both sera. PMID- 8425381 TI - Faster isolation of Brucella spp. from blood by isolator compared with BACTEC NR. AB - Brucella spp. was isolated from blood cultures in a mean time of 3.1 days (range, 2-5) in seven cases of human brucellosis processed with the Isolator lysis centrifugation system. Simultaneous BACTEC NR blood cultures in the same patients were positive in six, with a mean detection time of 20.6 days (range, 17-29). These data suggest a higher speed of recovery of Brucella spp. from blood specimens with the lysis-centrifugation procedure compared with the BACTEC NR system. PMID- 8425382 TI - In vitro activity of nine fluoroquinolone antibiotics against 200 strains of enterococci. AB - The in vitro activity of nine fluoroquinolones against 200 recent clinical isolates of enterococci was evaluated. The relative activity was WIN57273 > sparfloxacin > ciprofloxacin, temafloxacin > ofloxacin > fleroxacin, lomefloxacin, norfloxacin > enoxacin. Susceptibility to gentamicin or streptomycin did not influence the activity of the fluoroquinolones. PMID- 8425383 TI - Comparison of Streptex versus PathoDx for group D typing of vancomycin-resistant Enterococcus. AB - Vancomycin-resistant enterococci recently isolated from patient specimens were tested with both Streptex and PathoDx group D latex agglutination antisera. Using routine protocols, all isolates from patients at Temple University Hospital were positive with the PathoDx reagents and were negative with the Streptex reagents. Isolates required extensive boiling for positive reactions to occur using Streptex reagents. PMID- 8425384 TI - Attentional modulation of the phonetic significance of acoustic cues. AB - Four experiments addressing the role of attention in phonetic perception are reported. The first experiment shows that the relative importance of two cues to the voicing distinction changes when subjects must perform an arithmetic distractor task at the same time as identifying a speech stimulus. The contribution of voice onset time to phonetic labeling decreases when subjects are distracted, while that of FO onset frequency increases. The second experiment shows a similar pattern for two cues to the distinction between the vowels /i/ (as in "beat") and /I/ (as in "bit"). Under low attention conditions, formant pattern has a smaller effect on phonetic labeling while vowel duration has a larger effect. Together these experiments indicate that careful attention to speech perception is necessary for strong acoustic cues (voice-onset time and formant patterns) to achieve their full impact on phonetic labeling, while weaker acoustic cues (FO onset frequency and vowel duration) achieve their full impact on phonetic labeling without close attention. Experiment 3 shows that this pattern is obtained when the distractor task places little demand on verbal short term memory. Experiment 4 provides a data set for testing formal models of the role of attention in speech perception. Attention is shown to influence the signal-to-noise ratio in the phonetic encoding of acoustic cues; the sustained phonetic contribution of weak cues without close attention stems from reduced competition from strong cues. This principle is instantiated in a network model in which the role of attention is to reduce noise in the phonetic encoding of acoustic cues. Implications of this work for understanding speech perception and general theories of the role of attention in perception are discussed. PMID- 8425385 TI - Heuristics for scientific experimentation: a developmental study. AB - Scientific discovery involves search in a space of hypotheses and a space of experiments. We describe an investigation of developmental differences in the search constraint heuristics used in scientific reasoning. Sixty-four subjects (technically trained college students, community college students with little technical training, 6th graders, and 3rd graders) were taught how to use a programmable robot. Then they were presented with a new operation, provided with a hypothesis about how it might work, and asked to conduct experiments to discover how the new operation really did work. The suggested hypothesis was always incorrect, as subjects could discover if they wrote informative experiments, and it was either plausible or implausible. The rule for how the unknown operation actually worked was either very similar or very dissimilar to the given hypothesis. Children focused primarily on plausible hypotheses, conducted a limited set of experiments, designed experiments that were difficult to interpret, and were unable to induce implausible (but correct) hypotheses from data. Adults were much better than children in discovering implausible rules. The performance deficits we found were not simply the result of children's inadequate encoding or mnemonic skills. Instead, the adults appear to use domain-general skills that go beyond the logic of confirmation and disconfirmation and deal with the coordination of search in two spaces: a space of hypotheses and a space of experiments. PMID- 8425386 TI - A study of mental morbidity among primary care patients in Nigeria. AB - Psychological disorders among 272 primary care patients were assessed by a two stage screening procedure, using the 12-item version of the General Health Questionnaire (GHQ-12) and the Present State Examination schedule (PSE). The prevalence of psychiatric disorder was 21.3%. Depressive neurosis (51.7%) and anxiety neurosis (36.3%) were the most common disorders. The older age group, females, and those who were either widowed, separated, or divorced were found to be more likely to suffer from psychiatric morbidity. Primary health care (PHC) workers were only able to detect 13.8% of the psychiatric cases identified in the study population. The GHQ-12 was shown to be a feasible screening instrument for psychoemotional disturbance in the primary care setting (sensitivity, 83.7%; specificity, 79.8%; minimum misclassification rate, 19.4%). Based on these findings, short- and long-term training strategies are suggested for PHC workers in developing countries to help improve their abilities to detect and manage common mental disorders in primary care settings. PMID- 8425387 TI - European views on personality disorders: a conceptual history. AB - A historical account is offered of the European origins of the DSM-III-R category, personality disorder, and a differentiation is made between the history of terms, patterns of behaviors, and concepts. The concept of "disorder of character (personality)" developed in the nineteenth century, and was possible after notions such as character, constitution, temperament, and self received a psychological definition, and after the insanities became transformed into "psychoses." Terms such as "type" and "trait" were in turn ushered into the nineteenth century by faculty psychology and the phrenological tradition. Until the end of that century, "personality" referred to the subjective aspects of the self, and "personality disorder" meant alteration of consciousness (e.g., hysterical dissociation); the behavioral patterns dealt with by DSM-III-R as personality disorders were then called disorders of character and explained as states of volitional failure, or loss of coherence between cognitive, emotional, and conative functions, or of "automatism," i.e., the manifestation of lower (more primitive or animal) forms of behavior escaping the control of higher (human) ones. Character types and disorders were first considered as forms of attenuated insanity, and later on were made into a separate group whose membership could be attained via the visitation of a degeneration taint, neurological disease (e.g., encephalitis), or plain ill-breeding. The relevance of the intellectual background of the nineteenth century to these diagnostic changes is also discussed. PMID- 8425388 TI - A persuasion therapy for panic disorder in old Japanese medical literature. AB - This report presents two panic disorder cases reported in old Japanese medical literature and commentary on their psychotherapeutic approaches with reference to more recent theory. The first case was treated and described by Dr. Gen'yu Imaizumi in 1850, and the second case was reported by Dr. Masatake Morita in 1928. Both Imaizumi and Morita referred to their treatment as the "persuasion" method. They presupposed a vicious circle of escalating panic phobia, and made their patients face their fear of panic by implicitly paradoxical injunctions in order to break this cycle and free them from both their phobic anxiety condition and their repetitive panic attacks. PMID- 8425389 TI - The phenomenology of panic attacks in panic disorder with and without agoraphobia. AB - The frequency of symptoms during panic attacks and anticipation of the panic consequences were compared in patients with the subtypes of panic disorder (PD). Patients with moderate and severe agoraphobic avoidance reported that they had experienced more symptoms than patients with an uncomplicated PD (without agoraphobia [AG]); they also experienced almost all of the symptoms more frequently, with the difference being significant for a quarter of the examined panic symptoms. Panic patients with moderate and severe AG were also significantly more concerned about the loss of control and social and physical consequences of panic attacks. Taken together, these findings suggest that the severity of panic attacks, defined as the number of panic symptoms, along with a variety of anticipatory fears about the consequences of the attacks may contribute to the development of AG in panic patients. PMID- 8425390 TI - Presentation of acute psychosis in an Egyptian sample: a transcultural comparison. AB - The symptomatological and diagnostic differentiations and outcome of acute psychosis were studied in 50 Egyptian patients using the Schedule of Clinical Assessment of Acute Psychotic States (SCAAPS). The prevailing symptoms were delusions, worry, irritability, mood changes, and disturbed behavior. Sixty-four percent of the patients were symptom-free at 1-year follow-up assessment. Various factors that affect clinical and social outcome were discussed. The problem of diagnostic terms was also studied comparing SCAAPS terms with those of DSM-III-R and ICD-10. The most frequent diagnosis was psychogenic psychosis or brief reactive psychosis corresponding to the previously mentioned first and second systems, respectively. The inclusion of acute and transient polymorphic psychotic disorders with or without stress in ICD-10 will encompass those clinical syndromes in different cultures. A comparative study with a matched Indian sample was also conducted, the results of which are discussed in detail. PMID- 8425391 TI - Creutzfeldt-Jakob disease misdiagnosed as depressive pseudodementia. PMID- 8425392 TI - Comorbid features in bulimics before and after therapy: are they explained by axis II diagnoses, secondary effects of bulimia, or both? AB - This study evaluated the extent to which personality disorder (PD) diagnoses drawn in active bulimics explained pretreatment and posttreatment comorbid features, once possible secondary effects of bulimia nervosa (BN) on the mental status were controlled. We used structured interviews to assign axis II diagnoses to 73 DSM-III-R bulimics, and then examined whether or not classification into borderline PD (BPD), other PD (OPD), or no PD (NPD) groups (a) predicted pretreatment and posttreatment differences in comorbid symptoms, and (b) explained the severity of comorbid symptoms independent of effects attributable to BN sequelae. Groups never differed on eating-symptom measures, with all showing satisfactory average improvements over 3 months. However, on pretreatment and posttreatment comorbid symptoms, BPD subjects displayed more depression, reliance on maladaptive defenses, and other pathological features than did other groups. More importantly, after controlling for the effects of possible sequelae of BN on comorbid symptoms at each point in time, a borderline/nonborderline distinction always explained significant residual proportions of variance in comorbid symptoms. These findings suggest that despite tendencies for BN to exacerbate concurrent symptoms, axis II diagnoses remain meaningful as indicators of comorbid features. PMID- 8425393 TI - A controlled study of alexithymia in eating disorders. AB - The aims of the study were (1) to establish whether alexithymia is present in patients with bulimia nervosa (BN), (2) to compare bulimic patients with restricting anorexics (AN/R), bulimic anorexics (AN/BN), and normal controls with regard to alexithymia, (3) to determine whether in BN patients alexithymia is a state or a trait, and (4) to see whether alexithymia predicts short-term treatment outcome in BN. Study 1 included 173 eating disorder patients (BN: n = 93, AN/R: n = 55, AN/BN: n = 25) who were compared with 95 controls on the Toronto Alexithymia Scale (TAS). Study 2 included 41 BN patients who were assessed prospectively with the TAS before and after a 10-week drug treatment. AN/R patients in study 1 had significantly higher alexithymia scores than BN patients. All three eating disorder groups had significantly higher alexithymia scores than controls. For BN patients in study 2, TAS scores before and after drug treatment were stable, despite significant symptomatic improvement. We conclude the following: (1) eating disorder patients are considerably more alexithymic than normal controls; and (2) in BN, alexithymia may be a trait, unaffected by clinical improvement unless psychological treatment, encouraging the expression of emotions is offered. PMID- 8425394 TI - Comparison of screening proposals for somatization disorder empirical analyses. AB - We report a comparison of three sets of screening criteria, plus a post hoc modification of one, for the diagnosis of somatization disorder. Included in the comparison are the DSM-III-R seven-symptom screen, 11 symptoms identified by Swartz et al., and the hierarchical screen using symptom clusters that has been proposed for DSM-IV, along with a modification to the latter. Data sets used in this test were from the St Louis Epidemiologic Catchment Area (ECA) study and a family study of alcoholism. Findings of sensitivity, specificity, and positive predictive value indicate advantages and disadvantages of each screening set. No one set was clearly superior; selection of screening criteria should be dependent on the intended application. PMID- 8425395 TI - A direct comparison of the defense mechanisms of nondepressed people and depressed psychiatric inpatients. AB - This report presents a direct comparison of defensive styles (as measured by the Defense Mechanisms Inventory [DMI]) in a sample of depressed psychiatric inpatients and samples of nondepressed male and female normative groups. Consistent with the "depressive realism" literature, nondepressed men and women were more likely than their depressed counterparts to bias their perceptions in an overly cheerful, optimistic direction. Counternormative sex differences were also found. Depressed men were more likely to use internalizing defenses and depressed women were more likely to use externalizing defenses than their respective nondepressed comparison groups. Overall, as has been speculated, there was a relationship within depressed subjects between depression severity and the amount of negatively biased self-perception. PMID- 8425397 TI - Awareness of sexual harassment: first step toward prevention. PMID- 8425396 TI - Effects of depression and borderline personality traits on psychological state and eating disorder symptomatology. AB - The incidence of current or lifetime affective disorder and borderline personality characteristics were measured in bulimia nervosa patients. The relationship of these variables to the severity of eating disorder symptomatology (Eating Disorder Inventory [EDI]) and general psychiatric symptoms (Hopkins Symptom Checklist [SCL]) was examined. Categorical diagnostic assessments of affective disorder and borderline personality disorder (BPD) were made by Structured Clinical Interviews for DSM-III-R (SCID-I and -II). Affective disorder diagnosis (both current and lifetime) strongly influenced EDI and SCL profiles, while borderline personality characteristics had little influence. An understanding of the broad psychological symptomatology in bulimics requires the consideration of comorbid psychiatric illnesses, especially affective disorders. PMID- 8425398 TI - Taking the initiative when the doctor doesn't 'get it'. PMID- 8425399 TI - We must be willing to care for AIDS patients. PMID- 8425400 TI - Readers comment on response to chest tube instillation question. PMID- 8425401 TI - Readers comment on response to chest tube instillation question. PMID- 8425402 TI - Nursing care of patients receiving long-term infusion of neuromuscular blocking agents. PMID- 8425403 TI - Nursing care of patients receiving long-term infusion of neuromuscular blocking agents. PMID- 8425404 TI - Intramuscular administration of RATG in the heart transplant patient. PMID- 8425405 TI - Solving the organ donor shortage by meeting the bereaved family's needs. PMID- 8425406 TI - Immunosuppressive therapy after cardiac transplantation: teaching pediatric patients and their families. PMID- 8425407 TI - Renal transplantation: a case study of the ideal. AB - Although renal transplantation has become a commonly accepted treatment for ESRD, it is anything but routine for the patient and family. Although JB's was a real case, his course represents the ideal. Seldom is a case this uneventful and benign. Follow-up care is complicated, critical and individual, and nursing plays a crucial role at each step of the transplant process. PMID- 8425408 TI - What would cause the large V wave to come and go in monitoring pulmonary wedge pressure? PMID- 8425409 TI - What can a nurse do when the patient has an advanced directive and the physician disregards it? PMID- 8425410 TI - Teaching the teachers: creating an effective critical care preceptor program. AB - Staff nurse preceptors are integral members of effective critical care orientation programs. Much of the success of the orientation experience and orientee satisfaction depends on them. A critical care preceptor workshop containing a review of the orientation program and the application of educational principles in critical care settings has been found to be an effective method for assisting preceptors and for enhancing our orientation program. PMID- 8425411 TI - Pediatric critical care nurses' perceptions and understanding of cadaver organ procurement. PMID- 8425412 TI - Helping nurses to help patients and families in end-of-life decisions. Interview by Michael Villaire. PMID- 8425413 TI - Complications of acute myocardial infarction. PMID- 8425414 TI - Patterns of recurrence following high-dose preoperative radiation and sphincter preserving surgery for cancer of the rectum. AB - High-dose preoperative radiation with new sphincter-preserving surgical options for the management of distal and unfavorable cancers of the rectum is gaining recognition as an alternative to abdominoperineal resection and permanent colostomy. From 1976 to 1989, 161 patients with cancer of the rectum were entered into a program of high-dose preoperative radiation and radical sphincter preserving surgery. Selection was based on prospective clinical staging of unfavorability or tumor location at a low level in the rectum. All patients received a minimum dose of 4,000 to 4,500 cGy over 4 1/2 weeks in fractional doses of 180 to 250 cGy. Patients with tumor fixation were given an additional boost of 1,000 to 1,500 cGy for a total of 5,500 to 6,000 cGy using a coned-down field. Surgery was carried out four to eight weeks following completion of radiation. Fourteen patients, found at surgery to have liver metastasis, were treated by palliative resection. One hundred forty-seven patients underwent radical curative surgery with sphincter preservation. The surgical procedures performed were combined abdominotranssacral resection (63), transanal-abdominal transanal resection (53), and anterior resection (31). Follow-up ranged from a minimum of two years to 15 years, with a median of five years. There was no perioperative mortality. Anastomotic failure occurred in three patients, two of whom were reconstituted. Late diversion was required in 10 other patients, primarily for recurrent disease. One hundred thirty-four of the 147 patients (91 percent) maintained long-term normal sphincter function. Pelvic-perineal recurrence was observed in 18 patients (12.4 percent), 12 of whom had fixed tumors located below the 6-cm level of the distal rectum. Median time to local recurrence was 24 months. Distant metastasis with or without local recurrence occurred in 35 patients, 22 of whom had fixed tumors below the 6-cm level of the rectum. Median time to distant metastasis was 17 months. Forty-three patients have died, 32 of disease. The overall five-year Kaplan-Meier actuarial survival for the total group of patients is 79 percent, with a disease-free survival of 73 percent. The findings of this study indicate that high-dose preoperative radiation used in combination with radical sphincter-preserving surgical techniques results in excellent local control of disease, improved survival, and enhanced quality of life with retention of normal anal sphincter function. PMID- 8425415 TI - How accurate is endorectal ultrasound in the preoperative staging of rectal cancer? AB - In rectal cancer, depth of infiltration and metastatic involvement of lymph nodes are important prognostic factors. The correct choice of operative treatment depends on the extent of the disease. In a prospective study, the value of endorectal ultrasound in staging rectal cancer was evaluated, and factors affecting the method's accuracy are discussed. The overall accuracy in staging depth of infiltration was 89 percent. Overstaging occurred in 10.2 percent, understaging in 0.8 percent. Tumors of the lower rectum are incorrectly staged in 16.7 percent, whereas tumors of the middle and upper rectum had an incorrect staging in 6.3 percent (P < 0.001). Compared with computed tomography, endorectal sonography is the more accurate staging method (74.7 vs. 90.8 percent). In staging lymph nodes, the overall accuracy was 80.2 percent, sensitivity was 89.4 percent, specificity was 73.4 percent, positive predictive value (PPV) was 71.2 percent, and negative predictive value (NPV) was 90.4 percent. The staging accuracy depends on the size of the node. Endorectal ultrasound is a safe, inexpensive, and accurate staging method, in the assessment of both depth of infiltration and nodal status. The results are strongly related to the experience of the investigator. PMID- 8425416 TI - A simple and effective treatment for hemorrhagic radiation proctitis using formalin. AB - Radiation proctitis is a common complication of radiotherapy for pelvic malignancy. In the more severe form, it leads to intractable or massive hemorrhage, which may require repeated hospital admissions and blood transfusions. Medical therapy in patients with radiation proctitis is usually ineffective, whereas surgery is associated with a high morbidity and mortality. Eight patients (seven females and one male) with hemorrhagic radiation proctitis were treated over a six-month period with endoluminal formalin. The technique used ensured minimal contact with formalin. The median age of the patients was 68 years (range, 42-73 years). Seven patients had had cancer of the uterine cervix, and one patient had had cancer of the prostate treated with radiotherapy at a median time of 30 months (range, 9-46 months) previously. The median duration of time of symptomatic rectal hemorrhage before formalin therapy was eight months (range, 1-12 months). The median number of units of blood transfused previously per patient was four (range, 2-32). The time taken for formalin therapy was 20 minutes (range, 10-70 minutes). One patient required repeat formalin application at two weeks. Bleeding ceased immediately in seven patients after formalin treatment. No further bleeding was noted, nor was any blood transfusion needed, at follow-up at four months (range, 1-6 months). Formalin therapy is a simple, inexpensive, and effective treatment for hemorrhagic radiation proctitis. PMID- 8425417 TI - Evaluation and treatment of chronic intractable rectal pain--a frustrating endeavor. AB - A study was undertaken to assess the evaluation and treatment of chronic intractable rectal pain. Sixty consecutive patients, 23 males and 37 females with a mean age of 69 (range, 29-87) years and a mean length of symptoms of 4.5 years, were evaluated by questionnaire, office examination, anal manometry, electromyography, cinedefecography, and pudendal nerve study. In all cases, organic abdominopelvic and anorectal etiologies for the pain were excluded by extensive radiologic and endoscopic evaluation. All patients had failed conservative and medical therapy. Ninety-five percent of patients had one or more associated factors: constipation or dyschezia (57 percent), prior pelvic surgery (43 percent), prior anal surgery (32 percent), prior spinal surgery (8 percent), irritable bowel syndrome (10 percent), or psychiatric disorders (depression or anxiety; 25 percent). Possible etiologies for the pain included levator spasm or anismus in 62 percent, coccygodynia in 8 percent, and pudendal neuropathy in 24 percent of patients. Therapy for pain control included electrogalvanic stimulation (EGS) in 29, biofeedback (BF) in 14, and steroid caudal block (SCB) in 11 patients. Pain control was assessed by an independent observer at a mean of 15 (range, 2-36) months after completion of therapy. Continued successful pain relief was classified by patients as good or excellent after EGS in 38 percent, after BF in 43 percent, and after SCB in 18 percent; overall success was reported by 47 percent of patients. The presence of levator spasm, coccygodynia, or pudendal neuropathy did not influence outcome. The routine use of physiologic investigation of rectal pain may not be justifiable. Moreover, more than half of the patients were refractory to all three therapeutic options used in this study. PMID- 8425418 TI - Clinical and pathologic factors associated with delayed diagnosis in solitary rectal ulcer syndrome. AB - Solitary rectal ulcer syndrome (SRUS) often goes unrecognized or is misdiagnosed. Of 98 patients with a final clinicopathologic diagnosis of SRUS, an initially incorrect diagnosis had been made in 25 patients (26 percent). In these 25 patients with a misdiagnosis, the median age was 43 years and the female-to-male ratio was 3.2:1. The median duration of incorrect diagnosis was five years (range, three months to 30 years), and seven patients received prednisone (> 30 mg/day) for a mistaken diagnosis of inflammatory bowel disease. The main clinical symptoms were rectal bleeding (84 percent) and a disturbance of bowel function (56 percent). Rectal prolapse was present in 13 patients. Original rectal biopsy specimens from 23 patients were reviewed; inadequate specimens and failure to recognize diagnostic features of SRUS contributed to delayed diagnosis in 13 and 10 patients, respectively. The most common clinicopathologic misdiagnoses in SRUS patients with rectal ulcers or mucosal hyperemia were Crohn's disease and mucosal ulcerative colitis. In patients with "polypoid" SRUS, diagnostic confusion was usually with a neoplastic polyp. Persistent bowel symptoms and rectal lesions led to review of the presentations and repeat biopsy directed toward the edge of the rectal ulcers or from within the polypoid or hyperemic rectal lesions, finally establishing the diagnosis of SRUS. Intractable symptoms led to surgery in 15 patients (60 percent), with symptomatic improvement in over two-thirds. PMID- 8425419 TI - Protection of intestinal anastomoses by biodegradable intraluminal bypass tubes under the condition of general peritonitis: an experimental study on the CLP model in rats. AB - The purpose of this experimental study was to affirm the protective effect of biodegradable tubes for the intraluminal bypass procedure under the adverse condition of general peritonitis. General peritonitis was induced by means of the cecal ligation and puncture (CLP) model in the rat. The leakage rate in the control group (n = 20) without anastomotic protection was 70 percent (14/20). In three therapeutic groups, each consisting of 20 animals, the intestinal anastomoses were protected by an intraluminal bypass tube of different biodegradable biomaterials (collagen-II, BCL-002, and BCL-004). The best results were noted in the collagen-II and BCL-002 groups, where the leakage rates could be reduced to 10 percent. These highly significant results (P = 0.0001) prove the feasibility of biodegradable biomaterials for the intraluminal bypass procedure in the rat, even in cases with underlying peritonitis. PMID- 8425420 TI - Advanced colorectal neoplasia in the high-risk elderly patient: is surgical resection justified? AB - A review of the perioperative morbidity and mortality and long-term survival in elderly and high-risk patients with colorectal neoplasia was undertaken. Elderly high-risk patients with localized disease were compared with those with advanced disease. Over a five-year period, 82 high-risk (at least one major organ system disease), or elderly (age > or = 70 years) patients underwent an operation for colorectal neoplasia. Overall, 43 of 82 (52 percent) had advanced disease (obstruction, perforation, hemorrhage, or metastatic disease), while 39 of 82 (48 percent) had localized disease. The mean age of all patients was 78.2 years. Preoperative comorbid diseases included: coronary atherosclerosis, 59 (72 percent); previous myocardial infarction, 17 (21 percent); previous arrhythmia, 10 (12 percent); emphysema, 32 (39 percent); renal failure, 6 (7 percent); and cirrhosis, 3 (4 percent). At the time of surgery, 26 patients (32 percent) had metastatic disease. Six patients (7 percent) died in the perioperative period. The presence of advanced neoplasia did not significantly affect 30-day mortality. There was no difference in major morbidity between patients operated on for localized and for advanced disease. The mean actuarial 18-month survival was less for patients with advanced disease (P < 0.05). Sixty-eight patients (83 percent) are alive at a follow-up of 17.7 +/- 29 months postoperatively. The morbidity and mortality associated with resection of colorectal neoplasia in high-risk elderly patients are acceptable even in the presence of advanced disease. In select patients, resection offers the best palliation and may improve the quality of remaining life. PMID- 8425421 TI - Colon and rectal cancer in pregnancy. AB - Colorectal carcinoma presenting in pregnancy is an uncommon disease that is reported to be associated with an extremely poor prognosis. To better characterize this disease, we surveyed the membership of the American Society of Colon and Rectal Surgeons by mailed questionnaire and reviewed the literature. Forty-one new cases of women with large bowel cancer who presented during pregnancy or the immediate postpartum period were identified. The mean age at presentation was 31 years (range, 16-41 years). Tumor distribution was as follows: right colon-3, transverse colon-2, left colon-2, sigmoid colon-8, and rectum-26. Dukes stage at presentation was A = 0, B = 16, C = 17, and D = 6 (two patients were unstaged). Average follow-up was 41 months. Stage for stage, survival was found to be similar to patients with colorectal tumors in the general population. Large bowel cancer coexistent with pregnancy presents in a distal distribution (64 percent of tumors in the current series and 86 percent of those reported in the literature were located in the rectum) and presents at an advanced stage (60 percent were Stage C or more advanced at the time of diagnosis). While patient survival is poor, it is no different stage for stage from the general population with colorectal tumors. PMID- 8425422 TI - Functional intraperineal pouch of rectal wall (posterior rectocele). AB - We have identified 18 cases of functional pouch of the posterior rectal wall visualized by defecography. In the absence of previous evidence in the literature, we named this anomaly "posterior rectocele" and studied it from the clinical, radiologic, and endoscopic points of view. PMID- 8425423 TI - Anorectal dysfunction and rectal prolapse in progressive systemic sclerosis. AB - Our aim was to characterize the clinical spectrum of anorectal dysfunction among eight patients with progressive systemic sclerosis (PSS) who presented with altered bowel movements with or without fecal incontinence. The anorectum was assessed by physical examination, proctosigmoidoscopy, and anorectal manometry. There was concomitant involvement of the other regions of the digestive tract in all patients as determined by barium studies, endoscopy, or manometry: eight esophageal, three gastric, four small bowel, and two colonic. Seven patients had fecal incontinence, and four also had second-degree complete rectal prolapse. Abnormal anorectal function, particularly abnormal anal sphincter resting pressures, were detected in all patients; anal sphincter pressures were lower in those with rectal prolapse. Rectal capacity and wall compliance were impaired in seven of seven patients. Successful surgical correction of prolapse in three patients resulted in restoration of incontinence for six months and seven years in two of the three patients. We conclude that rectal dysfunction and weakness of the anal sphincters are important factors contributing, respectively, to altered bowel movements and fecal incontinence in patients with gastrointestinal involvement by PSS. Rectal prolapse worsens anal sphincter dysfunction and should be sought routinely as it is a treatable factor aggravating fecal incontinence in patients with PSS. PMID- 8425424 TI - Sigmoid volvulus in west Africa: a prospective study on surgical treatments. AB - To evaluate the efficacy of different types of surgery, we performed a prospective, randomized trial in 31 consecutively hospitalized patients with sigmoid volvulus. These patients represented 8 percent of 377 cases of emergency surgery. At the time of surgery, the patients were divided into two groups according to the absence (Group A) or presence (Group B) of bowel gangrene. At random, each group was assigned two surgical treatments. Seventeen patients entered Group A and underwent mesosigmoidopexy (seven patients) or resection and primary anastomosis (10 patients). Fourteen patients entered Group B and underwent Hartmann's procedure (eight patients) or resection and primary anastomosis (six patients). Overall mortality was four patients among 31 (13 percent), with a significant prevalence in the group with gangrene (21.4 percent vs. 5.8 percent). In Group A, the rate of success in patients treated with resection-anastomosis was higher than that in patients undergoing mesosigmoidopexy (90 percent vs. 71.5 percent). In Group B, a meaningful difference was observed between the rate of success of patients undergoing Hartmann's procedure and that of those undergoing resection and primary anastomosis (87.5 percent vs. 50 percent). The mortality rates were 12.5 percent and 33.3 percent, respectively. The results of our study show that the therapeutic approach to sigmoid volvulus should be diversified according to the absence or presence of gangrenous colon. The treatment of choice seems to be resection with primary anastomosis in patients with viable colon and Hartmann's procedure in patients with gangrenous colon. PMID- 8425425 TI - Complication of the intracolonic bypass. Report of a case. AB - The intracolonic bypass has been used both experimentally and clinically to avoid high-risk primary colonic anastomosis in the face of peritonitis. Experimental and clinical data have established the Coloshield as safe, with few clinical complications reported. This is a review of the literature and a case report of a complication of an intracolonic bypass that was found to have eroded through the colon in the early postoperative period. PMID- 8425426 TI - Success of estrogen-progesterone therapy in long-standing bleeding gastrointestinal angiodysplasia. Report of a case. AB - Gastrointestinal angiodysplasia is a common cause of occult bleeding. Surgical and endoscopic treatments are often ineffective. Recently, estrogen-progesterone therapy proved to be effective in these patients. We describe herein an 84-year old man who presented with prolonged gastrointestinal bleeding, in whom treatment with estrogen-progesterone stopped the bleeding. We suggest that hormonal therapy should be considered in cases of prolonged obscure gastrointestinal bleeding thought to be due to angiodysplasia. This therapy should be instituted intermittently for a prolonged period. The role of progesterone, especially in men, should be reconsidered. PMID- 8425427 TI - Reconstitution of intestinal continuity after extended left colectomy. AB - We wish to reintroduce an infrequently employed technique to re-establish intestinal continuity after extended resection of the left colon, transverse colon, and distal ascending colon. It involves bringing the proximal ascending colonic stump through the distal ileal mesenteric defect to reach the distal rectal stump in a tensionless fashion. PMID- 8425428 TI - Simple method of dilatation for strictured colostomy in children. PMID- 8425429 TI - Endoluminal transrectal ultrasonography: accuracy, reliability, and validity. AB - The significant risk of local recurrence after curative resection and the relative inaccuracy of preoperative clinical assessment justify a more sophisticated assessment for carcinoma of the rectum. Endoluminal rectal ultrasonography (EU) can directly visualize the degree of rectal wall penetration by tumor and the immediate pararectal lymph nodes. Despite several reports reporting excellent accuracy of EU compared with pathology in detecting the degree of tumor penetration, EU remains restricted in terms of widespread availability. A review of the literature was performed to determine the accuracy, reliability, and current validity of EU. Raw data were collected from cross sectional surveys assessing the degree of tumor penetration in 873 patients and lymph node involvement in 571 patients with primary rectal cancer. EU is very accurate at determining tumor penetration (kappa = 0.85) but is only moderately accurate at detecting lymph node involvement (kappa = 0.58). The reliability of EU has not been assessed, and a simple protocol is proposed. The validity of EU was assessed in only a few studies. EU is credible and feasible, makes intuitive biologic sense, and is, perhaps, sensitive to change. The ability to make clinical decisions based on EU (content validity) will decide whether widespread implementation is applicable. PMID- 8425430 TI - Management of acute incarcerated rectal prolapse. Correction. PMID- 8425431 TI - Chest pain as a consequence of abnormal visceral nociception. PMID- 8425432 TI - Biomechanical characteristics of the human esophagus. AB - Biomechanical wall properties of the human esophagus were studied. A probe, with a balloon designed for simultaneous measurement of cross-sectional area and intraluminal pressure, was placed in the esophagus 30 cm from the incisors. Tone was not detected before inflation of the balloon. When the balloon was inflated stepwise with pressures up to 40 cm H2O (30.7 mmHg), measurement of cross sectional area allowed calculation of distensibility and circumferential wall tension. Balloon cross-sectional area increased linearly with increased balloon pressure. Balloon distension induced contractions, both proximal to the balloon and at the site of distension, at a balloon pressure of about 15 cm H2O (11.5 mm Hg). The cross-sectional area for the threshold for distension induced contractions was 153 +/- 12 mm2 (diameter 14 mm). At the onset of these contractions, the contraction force was 15-20 cm H2O (11.5-15.3 mm Hg) and it increased to 47-58 cm H2O (36.1-44 mm Hg) at a balloon pressure of 20-40 cm H2O (15.3-30.7 mm Hg). Circumferential wall tension increased with increasing intraluminal pressure in an almost exponential manner. The pressure elastic modulus increased steeply at lower balloon pressures (10-20 cm H2O) (7.7-11.5 mm Hg), but at higher balloon pressures (20-40 cm H2O) (15.3-30.7 mm Hg) this increase was less. The circumferential wall tension and wall stiffness of the human esophagus increased with increasing balloon pressure and cross-sectional area. When a threshold is reached, distension induced contractions both proximal and distal to the balloon and at the distension site. PMID- 8425433 TI - Comparison of stationary vs ambulatory 24-hour pH monitoring systems in diagnosis of gastroesophageal reflux disease. AB - Twenty-four-hour intraesophageal pH monitoring is presently considered the most reliable diagnostic test for gastroesophageal reflux. Prolonged esophageal pH measurements can be obtained in hospitalized patients with a stationary technique and in ambulant outpatients by means of a portable device; however, there have been no studies that have examined whether the two approaches provide a similar diagnostic accuracy. We performed a prospective study to compare stationary and ambulatory pH-metry in the diagnosis of gastroesophageal reflux. Seventy-seven control subjects and 178 patients with proven gastroesophageal reflux disease were randomized to either ambulant or static pH-metry, which was performed with standard pH electrodes, sensors, and recorders. Reflux events (intraesophageal pH < 4.0) analyzed were: number of episodes; total, upright, and supine reflux time; number of episodes lasting > 5 min; and duration of the longest episode. A composite score of all reflux events according to DeMeester was also calculated. The limits of normality were defined as the 95th percentiles of the control groups. Both controls and patients assigned to either pH monitoring method were comparable. Of 255 studies attempted, 243 (95%) were completed successfully. The results showed similar median values of reflux events for the two control groups and for the two patients groups. Percent total reflux time provided a good separation between normal and abnormal reflux, with a sensitivity of 0.92 for static pH-metry and 0.68 for the ambulant procedure (respective 95th percentiles, 3.4 and 4.6).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425434 TI - Effect of graded exercise on esophageal motility and gastroesophageal reflux in trained athletes. AB - We evaluated the effect of graded exercise on esophageal motility and gastroesophageal reflux. We studied eight trained cyclists using a catheter with three strain-gauge transducers connected to a solid-state datalogger and an ambulatory intraesophageal pH monitor. Each study lasted 4 hr during which subjects exercised on a stationary bike for 1 hr at 60% of peak O2 uptake (O2 max), 45 min at 75% of O2 max, and for 10 min at 90% of O2 max. Subjects rested 1 hr before exercise (control period) and for 30 min between exercise sessions. Studies were performed after an overnight fast and subjects received only intravenous infusion of 5% glucose solution during the study. Plasma concentrations of gastrin, motilin, glucagon, pancreatic polypeptide (PP), and vasoactive intestinal peptide (VIP) were determined at rest and before and after each exercise session. The duration, amplitude, and frequency of esophageal contractions declined with increasing exercise intensity, and the differences were significant (P < or = 0.05) for all three variables at 90% O2 max. The number of gastroesophageal reflux episodes and the duration of esophageal acid exposure were significantly (P < or = 0.05) increased during exercise at 90% O2 max. Plasma hormone concentrations showed no significant changes between rest and the various exercise sessions. Thus, exercise has profound effects on esophageal contractions and gastroesophageal reflux which are intensity dependent. These effects are not mediated by the hormones measured. PMID- 8425435 TI - Does diet affect values obtained during prolonged ambulatory pressure monitoring. AB - With the development of a portable high-capacity data-recording device and fully automated computer analysis, it is now possible to monitor esophageal motility in an ambulatory outpatient setting and over a complete circadian cycle. However, limited data are available on the characteristics and pattern of esophageal motility in healthy subjects, particularly the effects of meals. We studied the effect of food types (liquid vs solid) and standardized vs nonstandardized diet on 17 healthy volunteers with a probe combining three miniature pressure transducers 5 cm apart. All subjects followed the same diet regimen: a standardized breakfast, strict liquid lunch, and no restriction for composition and quantity of dinner. The characteristics of contraction events (amplitude, duration, velocity, slope, area under curve) and their propagation types (peristaltic, simultaneous, segmental, retrograde) were analyzed and compared to supine and interprandial periods. The contraction characteristics and the propagation pattern were identical for the three types of meals. In comparison with the interprandial and supine periods, the three types of meals showed higher percent peristaltic contractions and smaller percent simultaneous contractions. The individual contraction characteristics were, however, not significantly different. Higher percentages of simultaneous, retrograde, and segmental contractions were found during the supine period than either the perprandial or interprandial periods. This study indicates that characteristics of esophageal contractions and propagation pattern are similar for meals of different composition and quantity. In comparison with interprandial and supine periods, the meals are always characterized predominantly by peristaltic contractions. Thus, standardization of meals during prolonged ambulatory pressure monitoring is not required. PMID- 8425436 TI - Epidemiology of hospitalization for achalasia in the United States. AB - Achalasia is an uncommon esophageal motility disorder of unknown etiology. To gain insights into possible etiologic risk factors, demographic and comorbidity data were obtained from Medicare hospital discharge data files from 1986-1989 on patients aged 65 and older. Age-adjusted sex- and race-specific occurrence rates were calculated for each US state. The rate of comorbid illness occurrence in achalasia patients was compared to that of the entire hospitalized Medicare population. Records of 15,000 achalasia discharges were available for analysis. Achalasia discharge rates increased linearly from age 65 to 94 years. They were similar in males and females as well as whites and nonwhites. High rates were observed in the South and low rates in most states of the East North Central region around the Great Lakes and in the Pacific region. The same geographic pattern was observed in men and women as well as in the two separate subsets of data representing the periods 1986-1987 and 1988-1989. Achalasia was associated with a significantly increased risk for pulmonary complications, malnutrition, and gastroesophageal cancer. The concordant occurrence of achalasia in patients with Parkinson's disease, depressive disorder, and various other myoneural disorders indicated a possible etiologic relationship. Achalasia appears to represent the clinical end point of several different pathways. Besides aging, different neurologic diseases may contribute to a loss in control of esophageal motility. The geographic pattern could suggest the influence of environmental factors. PMID- 8425437 TI - Prospective study of efficacy and safety of lansoprazole in Zollinger-Ellison syndrome. AB - Lansoprazole, a new substituted benzimidazole H+,K(+)-ATPase inhibitor, profoundly inhibits gastric acid secretion and has potential use in the management of diseases such as Zollinger-Ellison syndrome (ZES). In the present study we evaluated the efficacy and safety of lansoprazole in controlling acid hypersecretion in 20 patients with ZES. The starting dose was 60 mg once daily. Control of acid hypersecretion was defined as the dose required to reduce acid secretion to < 10 meq/hr in the last hour before the next dose. Doses were adjusted upwards until effective control was achieved. Patients not controlled with 120 mg once daily were placed on twice daily lansoprazole. Most patients (90%) required lansoprazole once daily. During long-term follow-up (mean 18.5 months), 25% of patients required upward dose adjustments and 25% of patients required twice daily lansoprazole. Following cessation of therapy, the mean time for gastric acid output to reach half basal acid output was 39.1 hr. Lansoprazole was well-tolerated without side effects. Clinical chemistry and hematological studies were unchanged, and no gastric carcinoids developed. These results demonstrate that lansoprazole is a safe and effective inhibitor of gastric acid hypersecretion in patients with Zollinger-Ellison syndrome. Because it has a long duration of action, lansoprazole can be used to control gastric acid hypersecretion in most patients with Zollinger-Ellison syndrome using a once daily dosing schedule. PMID- 8425439 TI - Epidermal growth factor, polyamines, and prostaglandins in healing of stress induced gastric lesions in rats. AB - Previous studies demonstrated that epidermal growth factor (EGF) and polyamines (PA) are capable of protecting gastric mucosa against topical irritants. This study was designed to examine whether EGF, PA, and PG affect the healing of acute gastric lesions induced by water immersion and restraint stress. It was found that the healing process of stress lesions in sham-operated rats was significant after 6 hr after stress, and after 24 hr the number of stress lesions was reduced by about 75%. In sham-operated rats, the healing of ulcerations observed at 6, 12, and 24 hr after the stress was accompanied by gradual restoration of DNA synthesis, and both these processes were significantly reduced by administration of DFMO (an inhibitor of ornithine decarboxylase activity) or indomethacin (an inhibitor of cyclooxygenase). In salivectomized rats, the healing was significantly delayed and the DNA was lowered at all time intervals after the stress. Administration of EGF, spermine, aminoguanidine (an inhibitor of degradation of PA), or 16,16-dmPGE2 after stress promoted significantly the healing and DNA synthesis, but pretreatment with DFMO abolished the effect of EGF but not that of spermine. We conclude that EGF, PA, and PG are implicated in healing of stress lesions and that EGF acts, at least in part, by the stimulation of PA formation in the gastric mucosa. PMID- 8425438 TI - The gastrointestinal tract in uremia. AB - Gastrointestinal mucosal abnormalities ranging from edema to ulceration occur in two thirds of patients dying of uremia. Early studies suggested that uremic patients on maintenance dialysis treatment were at increased risk of peptic ulceration but more recent data indicate that this is not so. Other gastrointestinal problems reported for uremic subjects on maintenance dialysis treatment include bleeding from telangiectatic lesions, constipation, mucosal deposition of amyloid and acute pancreatitis. Nausea and vomiting are common in the uremic patient but gastric emptying studies have yielded conflicting results. Patients undergoing renal transplantation are at increased risk of development of esophagitis, complicated peptic ulcer, intestinal ulceration, and perforation as well as acute pancreatitis. PMID- 8425440 TI - Endoscopy, gastric ulcer, and gastric cancer. Follow-up endoscopy for all gastric ulcers? AB - The practice of following benign-appearing gastric ulcers until healing was critically evaluated in a retrospective manner by reviewing all gastric ulcers that were followed with serial endoscopy and all gastric cancers diagnosed at the University of Alabama at Birmingham. The stated purpose of following ulcers to healing is to detect those gastric cancers that may be masquerading as benign ulcer and were not correctly diagnosed at initial endoscopy. Over a five-year period, 148 gastric ulcers were followed with serial endoscopy and in no case was an unsuspected carcinoma found at follow-up endoscopy. In addition, of 67 gastric cancers diagnosed between 1979 and 1986, 62 were suspected of being malignant by the endoscopist at initial examination for an accuracy of 92%. The accuracy rate based solely on biopsy and/or brush cytology was 94%. When endoscopic and biopsy and/or cytology impressions were combined, only one case of gastric carcinoma was not suspected. The overall accuracy was 99%. These results suggest that if either the endoscopic impression or the biopsy and cytology is suspicious for malignancy, then follow-up endoscopy until healing should be done. On the other hand, if, at the initial examination, the ulcer appears benign and biopsy plus cytology are negative, then serial endoscopy has a low benefit relative to its cost. PMID- 8425441 TI - Effects of cyclooxygenase and lipoxygenase inhibition on eicosanoids and healing of acetic acid colitis in rats. AB - Both cyclooxygenase products, such as prostaglandin (PG) E2, and lipoxygenase products, such as leukotriene (LT) B4, are increased in colitis and have potent proinflammatory actions. We studied effects of specific inhibitors of cyclooxygenase and 5-lipoxygenase on the healing of acetic acid colitis in rats. Acetic acid colitis was induced 24 hr before enzyme inhibition began. Four days after induction of colitis, the area of gross colonic mucosal damage was determined by image analysis. Eicosanoid content in the intestinal lumen was quantitated by radioimmunoassay following chromatographic purification. Under these conditions, indomethacin significantly retarded the healing of colonic lesions and inhibited PGE2 by > 90% compared to placebo-treated colitis rats. AA861 had no effect on the healing of lesions, although > 75% inhibition of leukotriene synthesis was demonstrated. These results suggest that inhibition of endogenous colonic prostaglandins can impair healing mechanisms in acute colitis even after inflammation has developed. In contrast, inhibition of leukotriene synthesis did not affect healing. PMID- 8425442 TI - Effect of chronic octreotide treatment on intestinal absorption in patients with acromegaly. AB - The adverse gastrointestinal effects of octreotide, a synthetic analog of somatostatin, have not been fully elucidated. Low-dose octreotide frequently causes adverse gastrointestinal symptoms in normal individuals. We investigated the adverse gastrointestinal effects of high-dose octreotide, which is required for the normalization of growth hormone hypersecretion in some patients with acromegaly. Patients with acromegaly (N = 8) were treated with octreotide, 450 micrograms/day, then 1500 micrograms/day for two months at each dosage. Carbohydrate absorption was assessed using the D-xylose test, and fat absorption using fecal fat excretion and serum carotene concentrations, at baseline, at each dosage of octreotide, and after one month of washout. Ultrasonography was used to monitor for cholelithiasis. Growth hormone and insulin-like growth factor-I concentrations were significantly suppressed at both dosages. Adverse gastrointestinal symptoms were mild and transient. D-Xylose absorption remained normal at each dosage and after one month of washout. Fecal fat excretion increased from 7 +/- 2 to 12 +/- 2 g/24 hr (P < 0.05) after the higher dosage and resumed baseline levels after the washout. Mean fasting serum carotene levels remained normal, and carotene loading test (15,000 units three times a day for three days) was unreliable in identifying patients with high fecal fat. No new cholelithiasis was detected by ultrasonography. One of two patients with preexisting cholelithiasis developed biliary colic several days after the treatment period. Although steatorrhea was common, small intestinal absorptive capacity was otherwise unchanged by four months of high-dose octreotide treatment, which significantly suppressed growth hormone secretion in acromegalic patients. PMID- 8425443 TI - Effects of PGE2, misoprostol, and enprostil on guinea pig enterocyte adenylate cyclase. Clinical implications. AB - Prostaglandins of the E series (PGE) are known to stimulate intestinal water and electrolyte secretions via the activation of the enterocyte adenylate cyclase. Their methylated synthetic analogs misoprostol and enprostil induced diarrhea in 5-13% of the patients in most clinical studies. In order to elucidate the role of PGE-adenylate cyclase interaction in these phenomena, we studied the stimulation of adenylate cyclase by native prostaglandin E2 (PGE2) and synthetic PGE analogs on isolated guinea pig intestinal epithelial cells. PGE2 stimulation of adenylate cyclase was dose-dependent, reaching a maximum for 3 x 10(-4) M, with an EC50 of 3.7 x 10(-6) M. The Hill analysis of the concentration-response curve gave a straight line, with a slope close to 1. The effect of PGE2 was strictly additive to that of 10(-5) M forskolin, whereas it was decreased in terms of potency by 10(-9) M cholera toxin. Somatostatin-14 markedly inhibited PGE2 stimulation by 37% and 45% with 10(-9) M and 10(-6) M, respectively. The two PGE methylated analogs misoprostol and enprostil were less potent than PGE2 in stimulating adenylate cyclase in our model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425444 TI - Potential usefulness of hydrogen breath test with D-xylose in clinical management of intestinal malabsorption. AB - Hydrogen breath tests (H2 BT) have been used extensively to investigate intestinal disaccharidase deficiencies. A potentially useful test for assessing intestinal absorptive function, the H2 BT with D-xylose (H2 BT-D-xylose), has received scant attention. We report here the results of our investigation of this test in 45 patients. Fifteen patients had proved malabsorption that was due to nontropical sprue in nine, and to lymphoma, Whipple's disease, or giardiasis in the remainder. Nine patients had small-bowel bacterial overgrowth secondary to either postsurgical sequelae or intestinal dysmotility. Twenty-one patients with irritable bowel syndrome and 21 healthy individuals served as control groups. All participants ingested 25 g of D-xylose, and alveolar breath samples were obtained thereafter at 30 min intervals for 5 hr. Breath H2 was measured by chromatography. Basal H2 production, peak change (delta) and area under the curve (AUC) were calculated. Simultaneously, 5-hr urinary excretion of D-xylose was measured by colorimetry and served as the reference test. In healthy individuals, D-xylose ingestion increased H2 production (delta = 5.8 +/- 1.4 ppm, P < 0.001). Changes were similar in patients with the irritable bowel syndrome. In contrast, the increase was of a much greater magnitude in the malabsorption group (delta = 49.9 +/- 7.2 ppm, P < 0.001 vs healthy controls). AUC analysis yielded comparable results. Test performance analysis showed that, in malabsorption the H2 BT-D xylose had a sensitivity index of 0.86, which was identical to that of the urinary D-xylose test. Specificity was 1 and 0.95, respectively; and predictability 1 and 0.93, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425445 TI - Respiratory methane excretion in children with lactose intolerance. AB - To evaluate the relationship between colonic methane production and carbohydrate malabsorption, we measured end-expiratory methane levels in 70 normal and 40 lactose-intolerant children. Time-dependent excretion of hydrogen and methane was determined every 30 min for 120 min following a fasting oral lactose challenge (2 g/kg). Mean breath hydrogen levels in normals (lactose-tolerant) equaled 3.7 parts per million (ppm) throughout the study, but increased to > 10 ppm by 60 min and remained elevated in lactose-intolerant subjects. Breath methane in normal children averaged 1.6 ppm from 0 to 120 min. In contrast, CH4 excretion by lactose-intolerant children averaged 5.1 ppm at 90 min; and, by 120 min levels increased significantly compared with control. Breath methane levels in lactose intolerant subjects following a lactose load continued to increase, however, despite the coingestion of exogenous lactase in amounts calculated to result in complete hydrolysis of the disaccharide. These data demonstrate that lactase deficient children manifest significant increases in breath methane excretion following lactose ingestion and that enhanced methane production may be a consequence of several factors, including altered fecal pH and increased methanogenic substrates provided by colonic lactose fermentation. Further studies are required to determine the clinical significance of elevated methane production in lactose intolerance. PMID- 8425446 TI - Morphologic alterations in small intestinal epithelium of lambs fed vitamin A depleted diet. AB - Ten crossbred wether lambs were fed once daily an oat diet that contained 3102 micrograms of retinyl palmitate (control) and were supplemented with 55,000 micrograms of retinyl palmitate orally once every two weeks. Twenty lambs were fed the same oat diet without retinyl palmitate supplements (A-depleted). After being fed the A-depleted diet for 28 weeks, 10 A-depleted lambs were repleted by feeding the control diet and oral supplementation of retinyl palmitate for eight weeks. The A-depleted lambs had serum vitamin A concentrations indicative of vitamin A deficiency, which was supported by very low liver vitamin A concentrations. Light microscopic and ultrastructural examinations revealed that alterations occurred at 50% and 75% of the small intestine length in A-depleted lambs only. Sawtooth configuration of the intestinal epithelium was a distinctive histologic feature. Consistent ultrastructural alterations were vesicular microvillar degeneration and disruption of the capillary endothelium. These results suggests that A-depleted diets have a detrimental effect on the small intestinal epithelium of lambs. Vitamin A repletion appears to minimize the detrimental effects. PMID- 8425447 TI - Characterization of secretin release in secretin cell-enriched preparation isolated from canine duodenal mucosa. AB - The release of secretin was studied in secretin cell-enriched preparations isolated from canine duodenal mucosa. The crude enterocytes were isolated by treating the duodenal mucosa sequentially with collagenase and ethylenediaminetetraacetic acid. Secretin cell-enriched fraction was prepared by centrifugation of the crude enterocytes in a counterflow elutriation rotor to obtain a final preparation containing 3.2 +/- 0.3 pmol/10(6) cell of immunoreactive secretin, which was 13-fold greater than the crude cell preparation (N = 5). The cells were incubated in Hanks' balanced salt solution for 20 min at 37 degrees C under 95% O2/5% CO2 before adding various agents and further incubated for various periods of time. The amounts of secretin released into the medium and retained by the cells were then determined by a specific radioimmunoassay. The release of immunoreactive secretin was increased dose dependently over the control by dibutyryl cyclic-3',5'-adenosine monophosphate, forskolin, 4 beta-12-O-tetradecanoylphorbol-13-acetate, the synthetic serine protease inhibitor, camostat, and the calcium ionophore, A23187. The effects of forskolin, the phorbol ester, and A23187 were time-dependent and not observed at 4 degrees C. The release of immunoreactive secretin was also stimulated by KCl in high concentration and by sodium oleate. The effect of A23187 was abolished in a Ca(2+)-free medium, while those of dibutyryl cyclic-3',5'-adenosine monophosphate and forskolin were potentiated by 3-isobutyl-1-methylxanthine, which did not have a significant effect when added alone. These results indicate that the release of secretin is regulated by both Ca(2+)- and cyclic-3',5'-adenosine monophosphate dependent mechanisms.2+ release. PMID- 8425448 TI - Scintigraphic studies of rectal emptying in patients with constipation and defecatory difficulty. AB - We prospectively evaluated 38 adult patients with chronic constipation with and without defecatory difficulties using a newly described scintigraphic test to measure rectal emptying and compared them to 20 healthy controls. All patients underwent anorectal manometry, and 30 who complained of infrequent defecation underwent a colonic transit study using radiopaque markers. Control subjects promptly evacuated both 100 ml and 200 ml artificial stool in a characteristic fashion, but three evacuated none of the 100-ml volume and two had no evacuation of the 200-ml stool (inhibited controls). Constipated patients exhibited three patterns of emptying: (1) normal emptying of both volumes (47%); (2) poor emptying of both volumes or inhibited defecation (29%); and (3) normal emptying of the 200-ml but abnormal evacuation of the 100-ml volume (24%). An abnormal expulsion pattern during manometry occurred in 21% of patients and was strongly associated with the inhibited defecation pattern. However, defecation patterns could not be predicted on the basis of age, gender, symptoms, duration of complaints, colonic transit, or other rectal manometric parameters. Although rectal scintigraphy has potential advantages as a diagnostic test in terms of quantitation and decreased radiation exposure, the inability of the test to distinguish patients with slow transit constipation and defecatory complaints makes the potential utility of this test of uncertain value in clinical and investigative settings. PMID- 8425449 TI - Octreotide, a long-acting somatostatin analog, in the management of postoperative dumping syndrome. An update. AB - Severe long-term complaints of dumping occur in a small number of patients after gastric surgery. Dietary modification, fiber preparations, and medical therapy are often ineffective. In these severely affected patients administration of the somatostatin analog octreotide before meals appears to be a promising new strategy. The effects of octreotide on both gastrointestinal transit time and hormonal changes appear to contribute to the benefits seen in dumping syndrome. However, as the majority of studies conducted have employed only a single dose of octreotide, careful long-term assessment of the nutritional and metabolic effects will be required. Recent results suggest that octreotide may be administered up to 2 hr before a meal and therefore has a sufficiently long duration of action to be of practical long-term use. Moreover, general improvements in life-style, as well as beneficial effects on symptoms, have been reported with long-term treatment, although the potential development of diarrhea will require careful monitoring. The development of an oral or nasal formulation should further improve the practical application of octreotide as a treatment for dumping syndrome. PMID- 8425450 TI - Regression of Barrett's esophagus by laser ablation in an anacid environment. AB - Consistent regression of intestinal metaplasia in Barrett's esophagus has not been achieved with medical or surgical interventions. In this case report, a patient with Barrett's esophagus of stable length had half the circumference of the Barrett's epithelium ablated with laser therapy while on a high-dose proton pump inhibitor. In the absence of esophageal acid exposure and after laser ablation, the intestinal metaplasia was documented to reepithelialize with normal squamous mucosa, which has persisted. PMID- 8425451 TI - Congenital esophageal stenosis presenting as noncardiac, esophageal chest pain. AB - A case of a 31-year-old female with congenital esophageal stenosis presenting with symptoms of chest pain caused by esophageal dysmotility is described. The involved segment in congenital esophageal stenosis has a characteristic thickening of the muscularis propria layer, as seen by EUS examination. In these patients, symptoms of dysphagia can be managed with esophageal dilation and noncardiac esophageal chest pain responds to pharmacotherapy with diltiazem. PMID- 8425452 TI - Endoscopic obliteration of recurrent tracheoesophageal fistula. AB - We report a successful endoscopic obliteration of a large recurrent tracheoesophageal fistula (diameter 0.6 cm, length 2.0 cm) in a 12-year-old girl, using a combination of Histoacryl (n-butyl-z-cyanoacrylate) and Aethoxysclerol injected through a polyethylene catheter. The severe pulmonary infection, which rendered surgery potentially life threatening, disappeared after the endoscopic closure. Since the obliteration, now over 12 months ago, the girl is asymptomatic. Endoscopic obliteration is a worthwhile technique and should be considered as an alternative to surgery in patients presenting with a complicated recurrent tracheoesophageal fistula. PMID- 8425453 TI - Is endoscopic sphincter of Oddi manometry a reproducible technique? PMID- 8425454 TI - Hypercholesterolemia in primary biliary cirrhosis is no risk factor for atherosclerosis. PMID- 8425455 TI - Can cyclosporin prevent proctocolectomy in severe steroid-refractory ulcerative colitis? Lessons from a case report. PMID- 8425456 TI - Severe acute pancreatitis following laser treatment of periampullary villous adenoma. PMID- 8425457 TI - [The prognostic importance of calcitonin screening in familial medullary thyroid carcinoma]. AB - In 34 of 139 patients (63 men, 76 women, mean age 41.2 +/- 15.6 years) with medullary thyroid carcinoma (MTC) the disease was of the familial variety. 13 of the 34 cases were discovered in the preclinical stage by screening (calcitonin stimulation with pentagastrin). This group was on average younger (16.8 +/- 10.7 years) than the 21 clinically manifest patients with MTC (28.0 +/- 11.9 years) or the 105 patients with sporadic MTC (45.7 +/- 13.6 years). The patients diagnosed through the screening programme had a more favourable tumour stage (12 in stage I or II) and were thus more likely to be cured. Their survival rate was higher (100% at both 5 and 10 years) than the survival rate of the patients with clinically manifest tumour (94 and 87%, respectively). But the group with manifest familial MTC did not differ from those with sporadic occurrence as regards the tumour stage (predominantly advanced stages), limited chance of cure and survival rate (5 years: 94%, 10 years: 81%). These data show that screening of family members decisively improves the prognosis of MTC of the familial variety. PMID- 8425458 TI - [Lethal Salmonella enteritidis meningoencephalitis in an adult with a carcinoma of an unknown primary site]. AB - A 71-year-old man with an aortobifemoral bypass graft had experienced several episodes of severe pain in the thoracic and lumbar spine, followed by severe abdominal pain, during the preceding two weeks. Physical examination, chest X ray, ultrasonography and computed tomography of the abdomen provided no specific pointers to a diagnosis. He was anaemic (haemoglobin 95 g/l), but had a normal WBC count with neutrophilia (up to 91%) and thrombocytopenia (up to 24,000/microliters). The transaminase and lactate dehydrogenase activities were raised, as were the bilirubin level and the retention values. There was a metabolic acidosis (pH 7.20). Because of suspected septicaemia ciprofloxacin (twice daily 200 mg) was administered intravenously. But the patient gradually lost consciousness, required mechanical ventilation and died 24 hours after admission in circulatory failure. Autopsy revealed a meningoencephalitis, carcinoma of unknown primary with bone marrow carcinomatosis as well as widespread carcinomatosis of blood and lymph vessels. The findings of blood, urine and sputum cultures, available only postmortally, revealed growth of tetracycline-resistant Salmonellae enteritidis. PMID- 8425459 TI - [Aorto-intestinal fistula versus angiodysplasia. A difficult differential diagnosis in gastrointestinal bleeding]. AB - A 63-year-old patient had recurrent tarry stools and haemoglobin levels of around 7.0 g/dl. Ten years previously he had undergone an aortobifemoral bypass operation for peripheral vascular disease in the legs. Eight gastroscopies and five coloscopies over ten weeks failed to discover a bleeding source. Although an aorto-intestinal fistula was considered early on, extensive diagnostic tests failed to reveal it. Digital subtraction angiography was suggestive of an angiodysplasia of the terminal ileum, a diagnosis supported when coloscopy during exploratory laparotomy visualized blood trickling from the terminal ileum. As a result of this finding a right hemicolectomy was performed. But recurrent bleedings necessitated relaparotomy which finally revealed a fistula between the ascending duodenum and the proximal bypass graft anastomosis. Nine months after resection of the proximal anastomotic area and interposition of a Dacron prosthesis the patient has been free of symptoms. PMID- 8425460 TI - [The hypotension risk during the therapy of heart failure with ACE inhibitors]. PMID- 8425461 TI - [The hemolytic-uremic syndrome and thrombotic thrombocytopenic purpura in adulthood. New aspects in their pathophysiology and therapy]. PMID- 8425462 TI - [Recruitment through industry-wide occupational medical services. The judgement of the Higher Regional Court of Munich of 16 April 1992]. PMID- 8425463 TI - [Right heart failure]. PMID- 8425464 TI - [Warming of erythrocyte concentrates?]. PMID- 8425465 TI - [Cholesterol concentration and cellular metabolism]. PMID- 8425466 TI - [Digitalis poisoning]. PMID- 8425467 TI - [The rectus abdominis syndrome]. PMID- 8425468 TI - [Mortality in familial hypercholesterolemia]. PMID- 8425469 TI - Fibroblast growth factors: what's in a name? PMID- 8425470 TI - Estrogen-induced calbindin-D 9k gene expression in the rat uterus during the estrous cycle: late antagonistic effect of progesterone. AB - Progesterone modulates estrogen-stimulated responses in the uterus. Calbindin-D 9k (CaBP9k), a 17 beta-estradiol-responsive gene expressed in the uterus, was used as a marker to examine the interactions between endogenous progesterone and estradiol in the rat. The variations in uterine CaBP9k messenger RNAs (mRNAs) during the rat estrous cycle indicated that CaBP9k gene expression was greatest during the estrogen-dominated phases (proestrus and estrus) and became totally repressed during diestrus, when progesterone predominates. Estradiol was found to be the major controlling factor of CaBP9k gene expression in vivo, progesterone antagonizing estrogen-induced CaBP9k gene expression. The inhibitory role of progesterone was further examined in two experiments. Mature cyclic rats were injected with the progesterone antagonist RU486 before the progesterone surge of proestrus, and the estrous cycle was mimicked in ovariectomized rats by sequential injections of estrogen and progestin. Progesterone did not appear to be involved in the rapid decrease in CaBP9k mRNA during estrus but was implicated in the down-regulation of the estrogen-stimulated CaBP9k gene expression at the end of estrus and during diestrus. This delayed effect of progesterone was confirmed in the ovariectomized rat model. CaBP9k mRNA accumulation in estrogen primed ovariectomized rats was suppressed by estrogen followed 1 h later by the progesterone agonist R5020. This effect occurred more than 24 h after progestin treatment. The inhibition of the estrogen-induced CaBP9k gene expression in the rat uterus by progesterone is certainly mediated by the progesterone receptor, because progesterone had no effect without estrogen priming or when the antagonist RU486 was used. The delayed progesterone effect probably does not involve depletion of nuclear estrogen receptors, the major rapid mechanism proposed for estrogen inhibition by progesterone in the rodent uterus, or control of estrogen receptor synthesis, as shown by Northern blot analysis of estrogen receptor mRNA. PMID- 8425471 TI - Discordant, organ-specific regulation of insulin-like growth factor-I messenger ribonucleic acid in insulin-deficient diabetes in rats. AB - Linear growth retardation is common in uncontrolled insulin-deficient diabetes, but individual organs such as kidney may hypertrophy. To explore whether this heterogeneity of response might be mediated by differential local insulin-like growth factor-I (IGF-I) gene regulation, we injected rats with ip saline, 65, 120, or 175 mg/kg streptozotocin (STZ). Diabetics were untreated or received daily insulin. Animals were killed 24, 48, or 72 h after documentation of diabetes, and liver, kidney, and lung messenger RNA (mRNA) content analyzed by solution hybridization/RNase protection assay. Untreated diabetics had 10- to 100 fold reductions in hepatic IGF-I mRNA apparent as early as 24 h, and the magnitude of these changes varied directly with the severity of diabetes. In contrast, kidney IGF-I mRNA content increased by 400-500% at 24 h in untreated diabetics given 175 mg/kg STZ, and by 100-200% at 48 h in those given 120 mg/kg STZ, with return to control levels by 72 h. Renal IGF-I mRNA levels actually decreased by 250-350% at 24 h in rats injected with 65 mg/kg STZ, returning to supranormal values by 72 h. These results suggest that severity and/or duration of the metabolic abnormality qualitatively and quantitatively affect this response in the kidney. Liver and kidney IGF-I mRNA levels approached normal with insulin therapy and were similar to controls in rats which received STZ but did not develop diabetes. Lung IGF-I mRNA levels were minimally altered in all experimental groups. At the time point and STZ dosage at which liver IGF-I mRNA changes were most dramatic, little change in liver alpha-tubulin mRNA was noted. At the time point and STZ dosages at which kidney IGF-I mRNA induction was most dramatic, renal IGF-I receptor mRNA was only minimally changed, and renal alpha tubulin mRNA was modestly reduced. In summary: 1) hepatic IGF-I mRNAs are dramatically reduced, and renal IGF-I mRNAs are significantly increased soon after the onset of insulin-deficient diabetes in STZ-treated rats; 2) insulin therapy restores IGF-I mRNA levels toward normal; and 3) these changes in IGF-I mRNA content are specific and are not the result of hepatic or renal STZ toxicity. These data suggest that IGF-I gene expression is regulated in a discordant, organ-specific manner in diabetes, and that metabolic factors in addition to GH may differentially modulate the endocrine and paracrine effects of IGF-I on growth. PMID- 8425472 TI - C-type natriuretic peptide mediates the hypothalamic actions of the natriuretic peptides to inhibit luteinizing hormone secretion. AB - Both A- and C-type natriuretic peptides (ANP and CNP, respectively) significantly reduce LH secretion when injected into the third cerebral ventricle of conscious rats. To establish which natriuretic peptide receptor subtype transduces these inhibitory messages, we have employed novel cytotoxin cell targeting techniques to selectively destroy cells in the hypothalamus that respond to ANP or CNP. Rats pretreated with ANP conjugated to the toxic A-chain of the plant cytotoxin ricin failed 1 week later to respond to central injection of ANP with the normal inhibition of LH secretion. These rats did, however, respond with significant inhibition of LH secretion to central injection of CNP. In fact, the LH inhibition observed after CNP injection was significantly greater than that expressed after similar injection of CNP in rats pretreated with unconjugated ricin A-chain (toxin control). Those control rats displayed significant reduction of LH levels in response to ANP injection as well. Plasma LH levels were not significantly affected by central administration of either ANP or CNP in rats pretreated with ricin A-chain conjugated to CNP. These results further demonstrate the power of this novel technology and provide positive evidence supporting our hypothesis that ANP exerts its LH-inhibiting effect by displacing endogenous CNP from clearance receptors within the brain. This endogenous CNP, then, like exogenously applied CNP, activates the guanyl cyclase-B receptors on cells, which are part of the network controlling the release of LHRH. PMID- 8425473 TI - Colocalization of immunoreactive endothelin-1 and neurohypophysial hormones in the axons of the neural lobe of the rat pituitary. AB - The intracellular localization of immunoreactive endothelin (ET)-1, and its colocalization with vasopressin (VP) and oxytocin (OT) in the rat neural lobe were investigated by immunogold techniques using specific antisera raised against ET-1, VP, and OT. There were two types of axons: the first contained VP immunolabeled neurosecretory granules, and the second contained OT-immunolabeled neurosecretory granules. A considerable number of the neurosecretory granules in both types of axon were immunolabeled with antibodies against ET-1, although the VP-immunolabeled granules were more heavily labeled with anti-ET-1 antiserum than OT-immunolabeled ones. Double immunogold labeling clearly demonstrated the intragranular colocalization of immunoreactions for ET-1 and VP and that of immunoreactions for ET-1 and OT. These results suggest that ET in the neural lobe may be released concomitantly with neurohypophysial hormones. Its biological significance remains to be elucidated. PMID- 8425474 TI - Chemical cross-linking of porcine luteinizing hormone: location of the cross-link and consequences for stability and biological activity. AB - To study the interaction between the subunits of LH and determine which amino acid residues are involved in this interaction, porcine and ovine LH (pLH and oLH) were cross-linked with 0.02 M 1-ethyl-(3-(3-dimethylaminopropyl)carbodiimide to generate one specific intersubunit cross-link. The cross-linked hormone was separated from the noncross-linked dissociated subunits by gel filtration on a Sephadex G-75 column. The position of the cross-link in cross-linked pLH (X-pLH) was determined by sequencing peptide fragments that were generated by digestion with endoproteinase Arg-C. In accordance with previous data for bovine LH, the position of the cross-link was between alpha-Lys49 and beta-Asp111, indicating that these residues are at the subunit-subunit interface. The biological activity of cross-linked hormone was tested by radioreceptor binding assay. The receptor binding activity of X-pLH was slightly reduced to 84%, suggesting that the conformational stability of X-pLH is similar to that of pLH as a result of the introduction of the covalent cross-link. The receptor-binding activity of X-oLH was decreased by approximately 30%, which we attribute to the formation of multiple cross-links within the ovine molecule, making the molecule more rigid, and to oligomeric forms, resulting from multiple intermolecular cross-links, that are not able to bind to the testicular LH receptor. This observation implies that the oLH has more amino-carboxyl functional groups that are sterically susceptible to carbodiimide cross-linking than does pLH under the reaction conditions used. PMID- 8425475 TI - Regulation of proteins in the cholesterol side-chain cleavage system in JEG-3 and Y-1 cells. AB - The conversion of cholesterol to pregnenolone, the rate-limiting step in steroid hormone synthesis, occurs on mitochondrial cytochrome P450scc, which catalyzes this reaction by receiving electrons from NADPH via a flavoprotein [adrenodoxin reductase (AdRed)] and an iron sulfur protein [adrenodoxin (Adx)]. The behavior of the genes and mRNAs encoding these proteins has been studied in several systems, but little is known about the behavior of the human proteins. Using cloned cDNAs for human P450scc and AdRed, we constructed bacterial expression vectors to make milligram quantities of the corresponding proteins. These, plus purified human Adx similarly prepared by Dr. L. Vickery, were injected into rabbits to raise antiserum to each of the proteins. Each antiserum was highly specific and did not cross-react with other mitochondrial proteins detectable by Western blotting. Human JEG-3 choriocarcinoma cells and mouse Y-1 adrenocortical carcinoma cells were then incubated for 0-24 h with 1 mM 8-bromo-cAMP (8Br-cAMP) or 30 nM phorbol 12-myristate 13-acetate (PMA; phorbol ester) plus 1 microM A23187 (calcium ionophore) to activate the protein kinase-A and -C pathways, respectively. In JEG-3 cells, 8Br-cAMP increased and PMA/A23187 slightly decreased the abundance of P450scc and Adx, but neither treatment had a detectable effect on AdRed. The production of pregnenolone by these cells increased 3-fold in response to 8Br-cAMP and fell to one third in response to PMA/A23187. In Y-1 cells, 8Br-cAMP increased the abundance of all three proteins, while PMA/A23187 decreased the abundance of P450scc and Adx. The production of pregnenolone by these cells increased 9-fold in response to 8Br-cAMP and was unaffected by TPA/A23187. These studies show that the three proteins of the cholesterol side-chain cleavage system behave in response to 8Br-cAMP and PMA/A23187 as predicted from the study of their genes and mRNAs, indicating that the chronic regulation of steroidogenesis in these cell systems is regulated principally at the level of mRNA abundance. PMID- 8425476 TI - Interleukin-1 stimulates the aromatase activity of human placental cytotrophoblasts. AB - Interleukin-1 (IL-1) is a multifunctional immunoregulatory peptide. Evidence suggests that IL-1 of either decidual or placental origin may serve a role in the paracrine/autocrine regulation of placental function, and the present studies were conducted to define the effects of IL-1 on the aromatase activity of human placental cytotrophoblasts. When freshly isolated cytotrophoblasts were incubated in medium supplemented with androstenedione, treatment with human IL-1 beta (hIL 1 beta) consistently increased the aromatization of this androgen to estrogens. In a representative experiment, hIL-1 beta (50 ng/ml) increased aromatization at 4 and 24 h by 145% (P < 0.001) and 78% (P < 0.05), respectively. This was indeed due to increased hIL-1 beta-mediated estrogen biosynthesis, rather than to decreased catabolism of estrogens, since cytotrophoblasts incubated in the presence of hIL-1 beta for 24 h exhibited 65% greater aromatase activity than control cells (P < 0.0001), as quantitated by the specific release of 3H2O from [3H] androstenedione. Stimulation of aromatase activity by hIL-1 beta was concentration dependent and saturable, and significant (P < 0.05) stimulation could be demonstrated at a hIL-1 beta concentration as low as 20 ng/ml. In time course studies, significant stimulation of the conversion of androstenedione to estrogens by hIL-1 beta could be detected as early as after 4 h of treatment and persisted for at least 24 h. Human IL-1 alpha stimulated the conversion of androstenedione to estrogens to an extent similar to that of hIL-1 beta, whereas the effects of murine IL-1 beta on aromatase activity were inconsistent. To determine whether endogenous IL-1 beta produced by cytotrophoblasts could itself act to stimulate aromatase activity, a neutralizing anti-IL-1 beta antibody was employed. When cells were incubated in the presence of anti-IL-1 beta antibody for either 4 or 24 h, the conversion of androstenedione to estrogens was significantly decreased compared to that in control incubations, whereas aromatase activity was not altered by the addition of nonimmune rabbit immunoglobulin G. These findings suggest that 1) both hIL-1 beta and hIL-1 alpha stimulate the aromatase activity of human cytotrophoblasts; and 2) endogenously produced IL-1 beta may function physiologically to enhance cytotrophoblastic aromatase activity. PMID- 8425477 TI - Prolactin receptors are found on heterogeneous subpopulations of rat splenocytes. AB - An increasingly large body of evidence implicates PRL as an immunoregulatory molecule. While most of the data relate to PRL levels and immunocompetence in vivo, we have shown that PRL is mitogenic for splenocytes from ovariectomized rats and rats in certain other hormonal states. This finding suggests that these lymphocytes express PRL receptors. Here, we wished to determine whether all or only a subset of splenocytes were PRL receptor positive. By using polyclonal as well as monoclonal antibodies to PRL receptor, we determined that as many as 20% of the primary splenocytes expressed PRL receptors. In a culture of Nb2 cells, a PRL receptor-positive lymphoid line, as many as 70% were PRL receptor positive. Dual labeling for lymphoid-specific antigen surface markers and PRL receptor indicated that about one third of the PRL receptor-positive splenocytes were kappa-light chain-positive B-cells, while the others stained with antibodies to T cell markers, CD4 or CD8. These data confirm that lymphocytes express PRL receptors and show for the first time that PRL receptor-positive lymphocytes are a heterogenous subset of total primary splenocytes. These cells may be the target for PRL-mediated immunoregulation. PMID- 8425478 TI - Androgens augment vasoactive intestinal peptide- and growth hormone-releasing hormone-stimulated progestin production by rat granulosa cells. AB - Vasoactive intestinal peptide (VIP) has been shown to stimulate steroid production by cultured rat granulosa cells independently of FSH. In the present study, we have examined the modulatory effects of various steroids on this response. Rat granulosa cells cultured for 2 days with only VIP showed small but significant increases in progesterone and 20 alpha-dihydroprogesterone (20 alpha OH-P) production. Concomitant treatment with either a synthetic estrogen (diethylstilbestrol), a synthetic progestin (R5020), or cortisol did not augment VIP-stimulated progesterone production; however, the latter two steroids slightly, but significantly, augmented VIP-stimulated 20 alpha-OH-P production. In contrast, concomitant treatment with a synthetic androgen (R1881) dramatically augmented both VIP-stimulated progesterone and 20 alpha-OH-P production. These effects were dose dependent for both VIP and R1881 and could be blocked by the androgen antagonist cyproterone acetate. Time course studies revealed that progesterone content of the culture media rapidly increased over the first 24 h of culture then remained fairly constant for the next 48 h; 20 alpha-OH-P content, on the other hand, was low for the first 12 h of culture and steadily increased thereafter. Dose-response analysis for R1881 revealed an ED50 of approximately 2 x 10(-8) M for the synthetic androgen, and comparison with other naturally occurring androgens provided the rank order of potency R1881 > androstenedione > testosterone = dihydrotestosterone. Additional studies with another member of the VIP peptide family, GH-releasing hormone, showed dose dependent stimulation of progesterone and 20 alpha-OH-P production by this peptide. These effects were also augmented by R1881 but not by diethylstilbestrol, R5020, or cortisol. These studies demonstrate that androgens, but not estrogens, progestins, or glucocorticoids, augment VIP- and GH-releasing hormone-stimulated progestin production by cultured rat granulosa cells. PMID- 8425479 TI - Native and modified (acetylated) low density lipoprotein-supported steroidogenesis by macaque granulosa cells collected before and after the ovulatory stimulus: correlation with fluorescent lipoprotein uptake. AB - We recently reported that uptake of fluorescent-tagged low density lipoprotein (DiI-LDL) by macaque granulosa cells (GC) was greatly enhanced within 27 h of an ovulatory stimulus (hCG injection). The present study was designed to determine whether increased DiI-LDL uptake correlated with an increased capacity for LDL supported steroidogenesis. We also tested whether modified [acetylated (ac)] LDL or high density lipoprotein (HDL), which are not ligands for the LDL receptor, supported progesterone (P) production. Beginning at menses, adult female rhesus macaques were treated with human (h) FSH and hLH for 9 days to promote development of multiple follicles. On day 10, monkeys were injected with hCG (1000 IU) or received no ovulatory stimulus. Large follicles were aspirated on day 10 (nonluteinized GC) or 27-34 h after hCG administration (luteinizing GC). GC (2 x 10(4)/0.2 ml) were cultured in Dulbecco's Modified Eagle's Medium-Ham's F 12 plus insulin, transferrin, H2SeO3, and aprotinin, with 0-100 micrograms hLDL, ac-hLDL, or hHDL. P concentrations in medium were determined by RIA. LDL (1-25 micrograms/ml) dose-dependently increased (up to 15-fold; P < 0.05) basal and hCG stimulated P production by luteinized GC on days 1-8 of culture. However, LDL (25 micrograms/ml) did not alter basal P production by nonluteinized GC and increased (2-fold; P < 0.05) hCG-stimulated P secretion only on days 4-8. Basal and hCG stimulated P production by luteinized GC were initially (days 1-2) increased (up to 2-fold; P < 0.05), but later (days 6-8) suppressed (P < 0.05) in a dose dependent manner by 1-100 micrograms ac-LDL/ml. Ac-LDL did not alter basal or hCG stimulated P production by nonluteinized GC. HDL (1-100 micrograms/ml) did not alter P production by either luteinized or nonluteinized GC. The number of viable luteinized GC on day 8 was reduced (30-50%; P < 0.05) after exposure to 10 micrograms ac-LDL/ml or more, whereas only the highest dose (100 micrograms/ml) of LDL or HDL reduced cell survival. Ac-LDL did not alter the survival of nonluteinized GC in culture. Flow cytometric analyses using fluorescent-tagged lipoproteins (DiI-LDL/DiI-ac-LDL) demonstrated the uptake of both native and ac LDL by luteinized GC. Uptake of DiI-LDL was competitively suppressed in a dose dependent manner by unlabeled LDL, but not by ac-LDL. In contrast, DiI-ac-LDL uptake was competitively inhibited by both ac-LDL and LDL.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8425480 TI - Endothelin-1 promotes steroidogenesis and stimulates protooncogene expression in transformed murine Leydig cells. AB - The effects of endothelin-1 (ET-1), a potent vasoconstrictor and mitogen to various cell types, on proliferation and differentiated functions of the murine Leydig tumor cell line MA-10 were investigated. Kinetic binding experiments at room temperature showed that [125I] ET-1 bound to MA-10 cells and reached equilibrium in 2 h. The data from competitive binding experiments yielded an apparent single class of high affinity binding sites characterized by a Kd and maximum binding capacity of 1 nM and 59 fmol/10(6) cells, respectively. For steroidogenic assays, cells were incubated with ET-1 (1 pM to 1 microM) and with epidermal growth factor (10 ng/ml) for 4 h at 37 C, and the progesterone levels in the medium were measured by RIA. Like epidermal growth factor, ET-1 caused about a 6-fold increase in progesterone production. ET-1 also enhanced the transient expression of the protooncogenes c-jun and c-myc by 3- and 2-fold, respectively. For proliferation studies, ET-1 (1 pM to 1 microM) was added to quiescent MA-10 cells for 24 h, and cell counts were performed; no increase in cell number was observed. The results of this study demonstrate that MA-10 cells possess high affinity binding sites for ET-1 and that ET-1 stimulates progesterone production and protooncogene expression, but not mitosis in this cell line. PMID- 8425481 TI - Rat liver 11 beta-hydroxysteroid dehydrogenase complementary deoxyribonucleic acid encodes oxoreductase activity in a mineralocorticoid-responsive toad bladder cell line. AB - The mineralocorticoid receptor displays equal affinity for aldosterone and corticosterone. It has been proposed that aldosterone selectivity in vivo is achieved by the conversion of corticosterone into its inactive metabolite 11 dehydrocorticosterone by 11 beta-hydroxysteroid dehydrogenase (11 beta HSD). To test this hypothesis, we transfected rat liver 11 beta HSD cDNA into TBM cells, a sodium-transporting cell line. These cells respond equally well to aldosterone and corticosterone, indicating that endogenous 11 beta HSD is expressed at low levels in TBM cells. Although exogenous rat liver 11 beta HSD was expressed at high levels in transfected cells, mineralocorticoid selectivity was not observed. By contrast, the biologically inactive 11-dehydrocorticosterone was readily converted into corticosterone, a potent agonist for sodium transport. Our results indicate that rat liver 11 beta HSD behaves predominantly as a reductase in TBM cells. Another 11 beta HSD isoform is likely to be responsible for the dehydrogenase reaction in aldosterone-responsive cells. PMID- 8425482 TI - Change in location and processing of inhibin alpha-subunit precursors during sexual maturation of the Djungarian hamster testis. AB - Immunohistochemical location and immunoblot of inhibin alpha-subunit peptides were analyzed in the testis of the Djungarian hamster from days 0-31 of postnatal development using a specific antibody. An intense immunoreaction was observed in the centrally located T1 prespermatogonia at day 0. The staining intensity decreased gradually in the spermatogonia when they make contact with the basal lamina at days 8-10. At days 13 and 15 there is no staining. Thereafter the immunoreactivity in Sertoli cells as well as in A spermatogonia gradually increased, being highest in sexually mature animals. The intensity of alpha subunit staining in the seminiferous tubules was stage specific, being strongest at stages III and IV. Immunoblot analysis of testis homogenates with the anti-INH alpha 1-32 antibody showed several bands: 88K, 80K, and 43K in immature hamster testis (0-, 2-, 6-, 8-, or 10-day-old). In the adult hamster (31-day-old) 88K, 80K, 28K, and 20K bands were seen, but no 43K band. Dimeric inhibin was not detected. The 43-44K band most likely corresponds to the pro-alpha N alpha C, the 28K band to intermediate forms between alpha N alpha C and alpha C (alpha I alpha C), and the 20K band to mature alpha-subunit (alpha C). The shift from the immature pattern to mature occurs at about 20 days of age. Freezing of the samples was deleterious to alpha C, since it could be detected only in freshly homogenized samples. The results suggest that prespermatogonia produce predominantly monomeric alpha-subunit precursor pro-alpha N alpha C, whereas the mature Sertoli cells as well as A spermatogonia contain mainly monomeric alpha I alpha C. The alpha-inhibin precursors may act as auto-/paracrine regulators of spermatogenesis. Our results suggest that different alpha-subunit precursors, pro alpha N alpha C and alpha I alpha C, might be involved in the differentiation and maintenance of spermatogenesis, respectively. The posttranslational processing of alpha-subunit precursors seems to play an important role in the physiology of reproduction. PMID- 8425483 TI - Chiroinositol deficiency and insulin resistance. I. Urinary excretion rate of chiroinositol is directly associated with insulin resistance in spontaneously diabetic rhesus monkeys. AB - Previously, we demonstrated that nondiabetic insulin-resistant monkeys had reduced covalent insulin activation of muscle glycogen synthase (GS) compared to normal monkeys and that covalent insulin activation of adipose tissue GS was absent in these monkeys. Covalent insulin activation of muscle and adipose tissue GS in monkeys with impaired glucose tolerance and noninsulin-dependent diabetes (NIDDM) was also absent. As in humans, monkeys with NIDDM have a lower urinary excretion rate of chiroinositol (CI), a component of a putative mediator of insulin action, compared to normal monkeys. To determine whether the urinary excretion rate of CI was related to insulin resistance, which develops naturally in many obese rhesus monkeys, we examined the relationships between 24-h urinary CI excretion rate and 1) whole body insulin-mediated glucose disposal rates (M) and insulin-mediated changes in 2) the skeletal muscle GS activity ratio (sm delta GSAR), 3) the skeletal muscle glycogen phosphorylase activity ratio, and 4) the adipose tissue GS activity ratio (at delta GSAR) in 27 monkeys ranging from normal (n = 12) to insulin resistant (n = 8) to overtly diabetic (n = 7). The urinary CI excretion rate was significantly correlated with M (r = 0.47; P < 0.02), sm delta GSAR (r = 0.38; P < 0.05), skeletal muscle glycogen phosphorylase activity ratio (r = -0.49; P < 0.01), and at delta GSAR (r = 0.46; P < 0.02). The urinary CI excretion rate was also correlated with glucose tolerance (r = 0.39; P < 0.05). There was a wide range of urinary CI excretion rates (0.42-5.17 mumol/day) in monkeys with normal fasting plasma glucose concentrations. However, of the 7 diabetic monkeys, 6 had a urinary CI excretion rate below 2.0 mumol/day, and in the subgroup of 16 monkeys with a urinary CI excretion rate less than 2.0 mumol/day, the associations of urinary CI with M rate (r = 0.65; P < 0.005), glucose tolerance (r = 0.63; P < 0.01), and sm delta GSAR (r = 0.73; P < 0.001) increased in strength and significance. We propose that the urinary CI excretion rate may be 1) a biochemical indicator of both in vivo and in vitro insulin resistance and 2) a noninvasive diagnostic tool with potential for the identification of those individuals at risk for NIDDM and other related diseases with insulin resistance. PMID- 8425484 TI - Chiroinositol deficiency and insulin resistance. II. Acute effects of D chiroinositol administration in streptozotocin-diabetic rats, normal rats given a glucose load, and spontaneously insulin-resistant rhesus monkeys. AB - The acute effects of administration of D-chiroinositol (D-CI), a component of a putative mediator of insulin action, on plasma glucose were examined in low dose streptozotocin-treated rats and normal rats given a glucose load and the effects on plasma glucose and insulin were determined in five obese rhesus monkeys with varying degrees of spontaneous insulin resistance. Single dose intragastric D-CI (10 mg/kg) administered to streptozotocin-treated rats produced a 30-40% decrease in plasma glucose (P < 0.05) at 30-120 min. Single dose intragastric D-CI (2-15 mg/kg) administered to normal rats 2 h before ip glucose produced a 30-50% decrease (P < 0.05) in plasma glucose. D-CI (10 mg/kg) caused a 50% increase (P < 0.05) in glucose disappearance rates in these rats. Myoinositol (10 mg/kg) was without effect. Intravenously administered single dose D-CI (100 mg/kg) increased both the glucose and insulin disappearance rates by 129 +/- 41% (mean +/- SE; P < 0.06) and 89 +/- 39% (P = 0.01), respectively, in all monkeys between 0-30 min compared to control values. D-CI administration, therefore, lowered elevated plasma glucose in streptozotocin-treated hyperglycemic rats, normal rats given a glucose load, and spontaneously insulin-resistant monkeys with or without noninsulin-dependent diabetes mellitus. Intravenous D-CI also lowered plasma insulin in these monkeys. PMID- 8425485 TI - Chiroinositol deficiency and insulin resistance. III. Acute glycogenic and hypoglycemic effects of two inositol phosphoglycan insulin mediators in normal and streptozotocin-diabetic rats in vivo. AB - Two insulin mediators, inositol phosphoglycans, were isolated from bovine liver by methods previously developed for rat liver, i.e. chromatography on an AG 1 x 8 ion exchange column and selective elution with HCl at pH 2.0 and 1.3. The pH 2.0 mediator containing D-chiroinositol stimulated pyruvate dehydrogenase phosphatase, whereas the pH 1.3 mediator containing myo-inositol inhibited cAMP dependent protein kinase. Each mediator was further purified by thin layer and Bio-Gel P4 column chromatography and injected ip into normal fed rats together with [U-14C]glucose. After 2.5 h, diaphragms were removed, and glycogen isolated. Insulin mediators, like insulin, stimulated [U-14C]glucose incorporation into glycogen by 150-160% in a dose-dependent manner in the nanomolar range. Mediators injected iv in the nanomolar range into low dose streptozotocin-diabetic rats decreased plasma glucose 30-45% in 30-60 min, with a return to basal concentrations after 150-180 min. These in vivo insulin-like effects of mediator were observed without changes in serum insulin concentrations. The pH 2.0 mediator was 50-100 times more active (per nmol organic phosphate) than the pH 1.3 mediator in the ip diaphragm glycogenesis assay. Mediator effects on diaphragm were completely blocked by preincubation with an immunopurified inositol phosphoglycan antibody. Both mediators were equally active iv in lowering plasma glucose (per nmol inositol) at concentrations comparable to those of insulin. PMID- 8425486 TI - Characterization of a human chorionic gonadotropin-like protein from Candida albicans. AB - Studies from our laboratory have demonstrated that human CG (hCG), human LH, (hLH), and an hCG-like protein extracted from Xanthomonas maltophilia were able to induce Candida albicans transition from the blastospore to the germ tube stage. In the present study, we describe the characterization of an hCG-like material extracted from Candida albicans blastospores (CaCGLP), which is potent in inducing transition and presumably represents the endogenous transition inducing substance. This material was extracted from Candida albicans blastospores with glacial acetic acid and purified by affinity chromatography using a polyclonal rabbit anti-hCG antibody. The product obtained is a 68 kilodalton single band protein, as analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot analysis. Under reduced conditions a protein smear is seen. Amino acid analysis showed a predominance of glycine (22%), followed by serine (12%), and glutamate (12%). This protein reacted in the following hCG immunoassays: 1) a polyclonal rabbit anti-hCG equilibrium assay, 2) a carboxyl-tail hCG equilibrium assay, 3) two hCG equilibrium assays using monoclonal antibodies (CG no. 4 and CG no. 9), 4) a free alpha-subunit equilibrium assay using a monoclonal antibody, and 5) an ultrasensitive immunoradiometric assay for hCG which does not cross-react with hLH, nor the free beta-subunit of hCG. The CaCGLP showed no reaction in a specific hLH immunoradiometric assay. When CaCGLP was tested in the transition assay, in the presence of 4% rat serum, it was found that this protein was 100 times more potent than hCG in producing Candida albicans transition. We conclude that Candida albicans produces a protein that has certain tertiary structure similarities to hCG and that this material is able to induce germ tube formation. We postulate that CaCGLP has an autocrine/paracrine effect in Candida albicans as a transition factor to control its own pathogenicity. PMID- 8425487 TI - Differential down-regulation of insulin-sensitive protein kinase-C isoforms by 12 O-tetradecanoylphorbol-13-acetate in rat adipocytes and BC3H-1 myocytes. AB - In rat adipocytes, chronic incubation with 12-O-tetradecanoylphorbol-13-acetate (TPA) reduced immunoreactive protein kinase-C (PKC) beta, gamma, delta, and zeta isoforms by 40-60% and PKC alpha by 75%, but had little effect on PKC epsilon levels. In BC3H-1 myocytes, chronic TPA treatment had no effect on PKC beta, increased PKC zeta, and depleted PKC alpha. Acute treatment with insulin induced the translocation of PKC beta in the myocytes both before and after chronic TPA treatment, but had no acute effect on the alpha or zeta isoforms. In contrast, acute TPA treatment in the myocytes had little effect on PKC beta, but induced the rapid translocation of alpha and zeta. The differential effects of chronic TPA treatment on the down-regulation of PKC beta may explain why insulin continues to activate biological processes in TPA-treated BC3H-1 myocytes, but not in adipocytes. PMID- 8425488 TI - The atrial natriuretic peptide system in rat ovaries. AB - The atrial natriuretic peptide (ANP) gene is expressed in several extracardiac tissues where ANP is thought to be involved in autocrine or paracrine regulation. The current studies were designed to characterize the ANP system in rat ovaries. ANP content in rat ovaries was estimated by RIA to be 240 +/- 70 pg/mg protein. HPLC revealed the presence of the 28-amino acid circulating peptide as well as the 126-amino acid prohormone, suggesting that the ovaries are a site of ANP synthesis. Indeed, ANP messenger RNA was detected in this tissue by RNase mapping. ANP present in ovarian extracts displaced [125I]ANP from bovine adrenal receptors (R1 class) in a dose-dependent manner and in parallel to the synthetic peptide, indicating that it possesses biological activity. Immunocytochemical studies localized ANP to interstitial cells surrounding the follicles; weaker but specific staining was also observed in the ovum. High affinity ANP receptors (dissociation constant, 0.30 +/- 0.06 nM; maximum binding capacity, 160 +/- 40 fmol/mg protein) were identified in ovarian membranes. Unlabeled ANP but not c atrial natriuretic factor (a specific agonist of ANP clearance receptors) competed with binding of [125I]ANP to ovarian membranes in a dose-dependent manner, suggesting that ovarian ANP receptors are predominantly of the R1 class. This was confirmed by cross-linking studies with [125I]ANP, which detected a single protein band with a molecular size of about 120 kilodaltons, corresponding to that of the guanylate cyclase-coupled R1 class of receptor. Consistent with the presence of biologically active receptors, ANP markedly enhanced cGMP accumulation (by 15-fold) in ovarian cells. The presence of both local ANP synthesis and high affinity transducing receptors in the ovaries indicates that the peptide plays a local role in ovarian growth or steroidogenesis. PMID- 8425489 TI - Hormone ontogeny in the ovine fetus. XXVII. Pulsatile and copulsatile secretion of luteinizing hormone, follicle-stimulating hormone, growth hormone, and prolactin in late gestation: a new method for the analysis of copulsatility. AB - To analyze the secretion patterns of LH, FSH, GH, and PRL in the late gestational sheep fetus in vivo, we measured simultaneous plasma levels of these hormones during a period of frequent sampling under basal conditions (samples every 15 min for 5 h) in 17 chronically catheterized sheep fetuses. To calculate mean plasma levels and areas under the curve, we analyzed hormone pulses and coincident pulse patterns to assess interactions between the release of these pituitary hormones. Mean plasma levels for all fetuses were: LH, 0.8 +/- 0.2 ng/ml (mean +/- SEM); FSH, 4.6 +/- 0.7 ng/ml; GH, 136.6 +/- 16.5 ng/ml; and PRL, 40.5 +/- 10.3 ng/ml. Pulse analysis detected 20 LH pulses during 5100 min of total sampling time, which gave a mean interpulse interval of 255.0 min. For GH, 37 pulses were detected; the mean interpulse interval was 129.7 min. Twenty PRL pulses yielded a mean interpulse interval of 225.0 min. FSH pulses could not be analyzed due to the long half-life of this hormone, but hormone level fluctuations were screened for maxima. A new method was developed to detect an interaction between hormone pulses. The probability of the simultaneous occurrence of hormone pulses was calculated and compared with the rate of coincidences found in the experiments. Analysis of copulsatile release of LH, GH, and PRL revealed 11 GH pulses coinciding with the LH pulses (P = 0.0020). An interaction between the pulsatile release of LH and GH can, therefore, be assumed. There was also a significant interaction between GH and PRL. Seven PRL pulses preceded the GH pulses by 15 min (P = 0.0014). In contrast, no significant copulsatile release could be observed between LH and PRL; 95.5% of LH pulses were accompanied by a maximum FSH level, suggesting an interaction between LH and FSH secretion. In summary, we show that LH, GH, and PRL (and possibly FSH) are secreted in a pulsatile fashion in the ovine fetus. Furthermore, the pulsatile releases of LH, FSH, and GH as well as GH and PRL are temporarily coupled, as demonstrated by a significant number of coincident pulses between LH/GH and GH/PRL and a high number of FSH hormone maxima concomitant with LH pulses. PMID- 8425490 TI - Pharmacokinetic profile of recombinant human (rh) inhibin A and activin A in the immature rat. I. Serum profile of rh-inhibin A and rh-activin A in the immature female rat. AB - The serum pharmacokinetics of recombinant human inhibin A (rh-inhibin A) and rh activin A were examined in immature female Sprague Dawley-derived rats after iv and sc injection of the drugs. After iv administration of rh-inhibin A (120 micrograms/kg), the serum concentrations were described by a biexponential equation. The weight-normalized clearance was 21.3 ml/min.kg, and the initial (t1/2 alpha) and terminal (t1/2 beta) half-lives were 2.9 min and 37.9 min, respectively. Subcutaneous administration of 120 micrograms/kg rh-inhibin A resulted in a peak serum concentration of 10.6 ng/ml at 30.8 min after injection. Approximately 24% of the sc administered material was absorbed. Serum concentrations of rh-activin A also declined biexponentially after iv injection of the drug (120 micrograms/kg). The clearance of rh-activin A was 5.1 ml/min.kg, the t1/2 alpha was 6.1 min, and the t1/2 beta was 46.3 min. The peak serum concentration of rh-activin A (104.7 ng/ml) was achieved 24.7 min after sc delivery of the drug. The bioavailability of the sc dose was 38%. Iodinated rh inhibin A and rh-activin A were used to examine the serum forms and metabolites of the drugs. [125I]rh-inhibin A and [125I]rh-activin A associated with two serum binding proteins. Within 2 min of iv injection, the labeled hormones bound follistatin and alpha-2-macroglobulin. Even though rh-inhibin A and rh-activin A are structurally similar and appear to bind to the same serum proteins, their disposition in the immature rat differ. PMID- 8425491 TI - Pharmacokinetic profile of recombinant human (rh) inhibin A and activin A in the immature rat. II. Tissue distribution of [125I]rh-inhibin A and [125I]rh-activin A in immature female and male rats. AB - The tissue distribution of recombinant human inhibin A (rh-inhibin A) and rh activin A was determined in immature female Sprague Dawley-derived rats after iv administration of radiolabeled proteins. [125I]rh-Inhibin A and [125I]rh-activin A diverge in their distribution to tissues of the immature female rat as examined histologically (whole body autoradiography and thin section analysis) and by computing the percent dose and tissue to blood ratios for individual tissues. [125I]rh-inhibin A accumulated in the spleen, adrenal, bone marrow, and ovary after iv injection. Iodinated rh-inhibin A was also found in the anterior and posterior pituitary. [125I]rh-activin A was found in the ovary and pituitary after iv injection. Little specific binding was found in the spleen or adrenal. The bone marrow accumulated some [125I]rh-activin A which was competed by rh activin A. The primary route of excretion for radioactivity was the kidney, with the label appearing in the bladder by 10 min after iv injection. Not only do rh inhibin A and rh-activin A have different pharmacokinetics, but fewer tissues accumulate radioactive rh-activin A than rh-inhibin A. PMID- 8425492 TI - Negative feedback effects of estrogen on luteinizing hormone-releasing hormone release occur in pubertal, but not prepubertal, ovariectomized female rhesus monkeys. AB - In a previous study we found that ovariectomy resulted in an increase in both LHRH release and LH release in pubertal monkeys but not in prepubertal monkeys. To determine whether this castration-induced LHRH increase is due to the removal of estrogen, in the present study, the effects of estradiol benzoate (EB, 30 micrograms/kg body wt) on in vivo LHRH release were examined using a push-pull perfusion method in prepubertal (age 15-19 months, n = 5), early pubertal (24-29 months, n = 5), and midpubertal (36-48 months, n = 5) female rhesus monkeys that were ovariectomized 3 to 5 months earlier. LHRH in 10-min perfusate fractions from the stalk-median eminence was measured from -6 to +24 h after EB injection. Circulating LH levels were also monitored over the same period at various intervals. EB decreased LH levels in early pubertal and midpubertal monkeys, whereas it did not cause any significant effects on LH release in the prepubertal monkey. EB also resulted in suppression of LHRH release in both early and midpubertal monkeys; mean LHRH release before EB in the early and midpubertal groups was 6.6 +/- 0.6 and 7.0 +/- 0.6 pg/ml.10 min, respectively. EB decreased mean LHRH release beginning 3 h after EB with the nadir occurring at 18-21 h after EB (1.0 +/- 0.2 pg/ml.10 min) in early pubertal monkeys and 21-24 h after EB (1.2 +/- 0.1 pg/ml.10 min) in midpubertal monkeys. Decrease of mean LHRH release was due to a decrease in LHRH pulse amplitude and basal release but not pulse frequency. Oil injection alone (control) failed to suppress LHRH and LH release. In contrast to the results in pubertal monkeys, mean LHRH release in prepubertal monkeys was not altered by EB (before EB, 1.1 +/- 0.2 pg/ml.10 min; 18-21 h after EB, 1.5 +/- 0.3 pg/ml.10 min). These results suggest that the LHRH neurosecretory system in pubertal monkeys is responsive to the negative feedback effects of estrogen. However, the fact that estradiol failed to suppress LHRH release in prepubertal monkeys indicates that the LHRH neurosecretory system and/or its regulatory systems are not sensitive to estradiol before the onset of puberty. These findings are consistent with the hypothesis that the increase in pulsatile LHRH release at the onset of puberty is not dependent on changes in ovarian steroid feedback mechanisms. PMID- 8425493 TI - Effects and interactions of prostaglandin F2 alpha, oxytocin, and cytokines on steroidogenesis of porcine luteal cells. AB - In the porcine corpora lutea (CL), prostaglandin F2 alpha (PGF2 alpha) and oxytocin (OXT) inhibit progesterone (P) but stimulate estradiol (E2) secretion from luteal cells kept under primary culture conditions. In vivo, both compounds are reported to have luteolytic properties when administered during the late luteal phase; in young CL, however, both substances stimulate P secretion, an effect which is E2-mediated. During the late luteal phase luteal cells appear to produce cytokines, and in addition, cytokine-producing macrophages invade the CL. We tested therefore whether cytokines, particularly tumor necrosis factor-alpha (TNF), have effects on basal or human CG-stimulated steroidogenesis. Furthermore, the interactions of cytokines with PGF2 alpha and/or OXT were investigated. TNF, and less potently interleukin (IL)-1 and IL-2 but not IL-6, inhibited basal as well as human CG-stimulated release of P and E2 in both small and large luteal cells. The inhibiting effect of PGF2 alpha and OXT on P secretion was augmented by these active cytokines. The stimulatory effect of PGF2 alpha and OXT on small and large luteal cell E2 production was completely inhibited. A profound stimulatory effect of E2 and small luteal cell P secretion was completely prevented by the cytokines, with TNF being more potent than IL-1 or -2. We conclude that the cytokines, particularly TNF, have luteolytic functions by their direct inhibiting effects on luteal cell P production. In addition, the cytokines inhibit synthesis and action of PGF2 alpha- and OXT-stimulated E2 secretion. Since E2 is a potent stimulator of luteal cell P production, this luteotropic signal is eliminated by cytokines, which add to the process of luteolysis. PMID- 8425494 TI - Endothelin-3 stimulates luteinizing hormone-releasing hormone (LHRH) secretion from LHRH neurons by a prostaglandin-dependent mechanism. AB - Endothelin (ET) peptides have recently been recognized as putative regulators of the endocrine system. Particularly in the gonadal system, ET-3 stimulates LH secretion from anterior pituitary cells cultured in vitro. In these studies, we evaluate the actions of ET-3, the most abundant species of the ET family in the central nervous system, on LHRH release from arcuate nucleus-median eminence (AN ME) fragments and an LHRH-secreting neuronal cell line (GT1 cells) in vitro. ET-3 exhibited a stimulatory effect on LHRH secretion from AN-ME fragments and GT1 cells incubated in a static system as well as in a dynamic perifusion paradigm. In all the systems used, the effects of ET-3 on LHRH secretion showed a dose dependency. The increase in LHRH secretion induced by ET-3 was accompanied by an increased secretion of prostaglandin E2 (PGE2), not only in the AN-ME incubations, but also in the GT1 incubation and perifusion systems. Blockade of arachidonic acid and/or PG synthesis significantly reduced the ET-3-induced LHRH and concomitant PGE2 release from both AN-ME fragments and GT1 cells incubated in vitro. In AN-ME incubations, ET-3 effects were enhanced by potassium-induced depolarization. This suggests that activation of other transmitter system(s) may be needed for obtaining a physiological activation of the LHRH neuronal system. In summary, in these studies we provide evidence for a direct action of ET-3 on the LHRH neuronal system. This action is exerted directly on LHRH neurons either at the level of the nerve terminals, the perikaryon, or both. In addition, the effects of ET-3 on LHRH release require a functional arachidonic acid metabolic pathway, particularly involving PG synthesis, in order to obtain stimulation, indicating that PGs are involved in the intracellular events leading to ET-3 evoked LHRH secretion. PMID- 8425495 TI - The 22K variant of rat prolactin: evidence for identity to prolactin-(1-173), storage in secretory granules, and regulated release. AB - Western blot analyses of the rat pituitary have detected a 22K PRL variant distinct from intact PRL (25K). We recently reported that glandular kallikrein (GK), an estrogen-induced lactotroph protease, can process PRL in vitro from a 25K form to a 22K form in a thiol-dependent cleavage at Arg174-Arg175 to remove 23 amino acids. We also detected an estrogen- and thiol-induced 22K PRL variant in the rat pituitary comigrating with a PRL product generated by in vitro processing with GK and carboxypeptidase-B. This study addressed whether the in vivo 22K PRL variant originates through a GK-like cleavage and is a regulated secretory product of the rat pituitary. A polyclonal antipeptide antiserum was raised against a synthetic peptide [PRL-(163-173)] representing the new C terminus after GK and carboxypeptidase-B processing. In slot and Western blots, this antiserum (CT-antiserum) specifically recognized PRL processed in vitro by GK and carboxypeptidase-B and did not recognize intact PRL or PRL cleaved by GK alone. Western blot analysis of rat pituitary extracts with CT-antiserum specifically detected an estrogen- and thiol-induced 22K band that comigrated with a PRL product generated by in vitro processing with GK and carboxypeptidase B. This 22K band was concentrated in subcellular fractions of the pituitary enriched in secretory granules. During short term incubations in medium 199, pituitaries from normal adult female rats released substantial amounts of 22K PRL; in contrast, male pituitaries did not release detectable 22K PRL. The release of 22K PRL from female pituitaries was powerfully blocked by bromocriptine, a dopaminergic agonist. GK was also released from pituitaries of female, but not male, rats, and GK release was inhibited by bromocriptine. The results identify the 22K PRL variant as PRL-(1-173), which is consistent with GK like processing at Arg174-Arg175, followed by carboxypeptidase-B-like processing. The results also show that 22K PRL is a natural female-specific secretory product of the rat pituitary under inhibitory dopaminergic control. PMID- 8425496 TI - Functional and metabolic perturbations in isolated pancreatic islets from the GK rat, a genetic model of noninsulin-dependent diabetes. AB - Spontaneously diabetic nonobese GK rats exhibit high basal plasma glucose and insulin levels and a poor insulin secretory response to glucose. We studied insulin biosynthesis, insulin release, and glucose metabolism in freshly isolated islets from GK rats and control Wistar rats. In GK rats, islet insulin content was decreased when expressed per islet but normal when related to DNA content. In the presence of a low concentration (2.8 mM) of glucose both (pro)insulin and total protein biosynthesis was doubled in islets from GK rats. As judged from the (pro)insulin/total protein synthesis ratio, (pro)insulin biosynthesis was normally stimulated by 16.7 mM glucose. In islets from diabetic rats both basal and glucose-stimulated insulin release were dramatically decreased. A reduced secretory response to 10 mM leucine or 10 mM leucine plus 10 mM glutamine and a lack of response to 10 mM monomethylsuccinate were observed. By contrast the insulinotropic capacity of nonnutrient secretagogues such as 62 microM gliclazide or the combination of 2 mM Ba2+ and 1.4 mM theophylline in the absence of extracellular Ca2+ remained normal. Glucose oxidation (estimated as the production of 14CO2 from D-[6-14C]glucose) was severely impaired, whereas no major alteration of glycolytic flux (as judged from the conversion of D-[5 3H]glucose to 3H2O) could be detected. Accordingly, the D-[6-14C]glucose oxidation/D-[5-3H]glucose use ratio was less markedly increased in response to a rise in glucose concentration in islets from GK rats than in islets from control rats. Thus, in islets from diabetic GK rats, glucose-induced insulin release but not insulin biosynthesis was impaired. This defect is associated with, and probably at least in part due to, a deficient oxidative metabolism of glucose in islet mitochondria. PMID- 8425497 TI - Parathyroid hormone is more effective than estrogen or bisphosphonates for restoration of lost bone mass in ovariectomized rats. AB - The study was designed to compare the therapeutic efficacy of estrogen, the bisphosphonate risedronate (NE-58095), and PTH for restoration of lost bone mass in osteopenic, ovariectomized (OVX) rats. In addition, the skeletal effects of these single treatments were compared to those of concurrent treatments with PTH + estrogen or PTH + NE-58095. OVX rats were untreated for the first 4 weeks postovariectomy to allow for the development of moderate tibial osteopenia. These animals were then subjected to the various treatments for periods of 5, 10, and 15 weeks. Their proximal tibiae were processed undecalcified for quantitative bone histomorphometry. Treatment of osteopenic OVX rats with estrogen or NE-58095 alone depressed bone turnover and prevented additional cancellous bone loss from occurring during the treatment period. However, these therapeutic agents failed to restore lost bone in OVX rats to control levels. In contrast, OVX rats treated with PTH alone exhibited a marked stimulation of bone formation which resulted in augmentation of cancellous bone mass to a level 2-fold greater than that of vehicle-treated control rats. Concurrent treatment of OVX rats with PTH + estrogen as well as PTH + NE-58095 also effectively reversed cancellous osteopenia in OVX rats, but did not appear to be more beneficial to the estrogen deplete skeleton than treatment with PTH alone. The results indicate that PTH is a powerful stimulator of bone formation and completely restores lost cancellous bone in osteopenic OVX rats. Furthermore, the bone anabolic effects of PTH are much more pronounced than those of estrogen or bisphosphonates. These findings in an animal model of estrogen depletion provide support for PTH as a potentially effective treatment for oophorectomized and postmenopausal women with established osteoporosis. PMID- 8425498 TI - 1,25-Dihydroxyvitamin D3 and phorbol myristate acetate produce divergent phenotypes in a monomyelocytic cell line. AB - The human monomyelocytic HL-60 cell line differentiates along a monocytic lineage when cultured in the presence of phorbol myristate acetate (PMA) or 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3]. The protooncogene c-fms, coding for the macrophage colony-stimulating factor (M-CSF) receptor, is characteristically expressed in cells of monocytic lineage, but is not present in HL-60 cells. Since M-CSF has an undefined role in osteoclast development, we wished to examine the effect of 1,25-(OH)2D3, a strong promoter of osteoclast development, on the induction of M-CSF receptor. Treatment of HL-60 cells with 10-50 ng/ml PMA for 3 days stimulated expression of c-fms, as measured by steady state mRNA levels. Treatment with 10 nM 1,25-(OH)2D3 for 4-72 h did not promote c-fms expression. In fact, 1,25-(OH)2D3 attenuated PMA-stimulated c-fms expression in a dose-dependent manner (ED50, 1 nM). Along with attenuation of c-fms expression, 1,25-(OH)2D3 induced expression of mRNA for the osteoclastic enzyme carbonic anhydrase-II (CA II) almost 2-fold over the level expressed in untreated HL-60 cells. The addition of PMA to the culture diminished the basal expression of CA II mRNA, but did not affect 1,25-(OH)2D3-stimulated CA II induction. PMA and 1,25-(OH)2D3, thus, promote divergent phenotype development in the HL-60 cell. Induction of c-fms mRNA by PMA should ensure further macrophage development. The effect of 1,25 (OH)2D3 to attenuate expression of c-fms mRNA and stimulate CA II mRNA suggests that 1,25-(OH)2D3 activates osteoclast, rather than macrophage, development. 1,25 (OH)2D3-induced phenotypic changes may, therefore, involve modulation of the M CSF effect. PMID- 8425499 TI - Ontogenesis of iodothyronine deiodinase activities in brain and liver of the chick embryo. AB - The ontogeny of 5'-monodeiodinase activity (5'-MA) was analyzed in chick embryo brain and liver tissue. The enzymatic type predominant in this path and the activity of the deactivating pathway (5 MA-III) were determined during certain periods of neural development in both organs. Results show that T3 is predominantly formed in both organs during the first third of embryogenesis (from day 5) until neuroblast proliferation (day 13). Within this lapse, the slow type II enzyme (insensible to propylthiouracil) is present in the brain, whereas in the liver the predominating enzyme is type I (the rapid enzyme). Further along the synaptogenesis period (day 14-17), 5' deiodination virtually disappears in the brain, the hepatic type I enzyme switches to the slow autoconsume enzyme (type II), and 5 MA-III levels increase significantly in both organs. Finally, on days 18-20 (perinatal period) the 5' pathway reaches the highest levels observed throughout the study in both tissues. Associated to this increase, liver enzymatic activity returns to type I. During this period, 5 MA-III is reduced by 40% in the brain and disappears from the liver. Together, these data strengthen the notion of a protective mechanism against brain overexposure to T3 during synaptogenesis and suggest that the protective mechanism also involves the regulation of extraneural deiodinases. PMID- 8425500 TI - Effect of homologous placental lactogens, prolactins, and growth hormones on islet B-cell division and insulin secretion in rat, mouse, and human islets: implication for placental lactogen regulation of islet function during pregnancy. AB - Up-regulation of maternal islet function is essential to accommodate the increased demand for insulin during pregnancy. Previously, we suggested that lactogenic activity regulates islet function during pregnancy. However, this hypothesis was based on the effect of homologous PRLs on islets, since the homologous placental lactogens (or islets) were unavailable. In this study we examine the direct effects of homologous placental lactogens (PL), PRL, and GH on insulin secretion and B-cell division in rat, mouse, and human islets in vitro. Neonatal rat islets were cultured for 8 days in the presence of 0-1000 ng/ml rat PL-I (rPL-I), rPRL, or rGH. Media were changed daily, and the insulin concentration was determined. rPL-I and rPRL (500 ng/ml) treatment resulted in a 2-fold increase in insulin secretion. rGH (1000 ng/ml) elicited a 30% increase in insulin secretion. Similarly, cell replication, as indicated by BrdU incorporation into B-cells, was increased 4-fold in the presence of rPL-I and rPRL. The ED50 for insulin secretion and 5'-bromo-2'-deoxyuridine (BrdU) incorporation was 70 ng/ml for rPL-I and 150 ng/ml for rPRL. Similarly, in adult rat islets, insulin secretion was increased 1.6-fold, and B-cell replication increased 3-fold in the presence of the lactogenic hormones. Neonatal mouse islets were cultured for 8 days in the presence of 500 ng/ml mouse (m) PL-I, mPL II, mPRL, or mGH. mPL-I, mPL-II, and mPRL treatment resulted in a 2-fold increase in insulin secretion. mGH elicited a 30% increase in insulin secretion. BrdU incorporation into B-cells was increased 3-fold in the presence of mPL-I and mPRL and 2-fold in the presence of mPL-II. Adult human islets were cultured for 8 days in the presence of 1 microgram/ml human (h) PL, hPRL, or hGH. For human islets isolated from six pancreata obtained from females, hPL (138 +/- 10%), hPRL (133 +/- 9%), and hGH (117 +/- 3%) significantly increased insulin secretion compared to that from control islets. This study compares the direct effects among homologous PLs, PRLs, and GHs on insulin secretion and B-cell division in rat, mouse, and human islets. The results indicate that placental lactogen directly regulates islet function in several species and is probably the principal hormone responsible for the increased islet function observed during normal pregnancy. PMID- 8425501 TI - Levels of messenger ribonucleic acid for cholesterol side-chain cleavage cytochrome P-450 and 3 beta-hydroxysteroid dehydrogenase in bovine preovulatory follicles decrease after the luteinizing hormone surge. AB - During the follicular/luteal phase shift in steroidogenesis, follicular steroid production changes from predominantly estradiol and androgen secretion before the LH surge to decreased androgen and estrogen and increased progesterone after the LH surge. Our objective was to determine whether changes in progesterone production by the preovulatory follicle are effected via changes in mRNA levels for the steroidogenic enzymes cholesterol side-chain cleavage cytochrome P450 (P450scc) and 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4-isomerase (3 beta HSD). Bovine preovulatory follicles were obtained in the early follicular phase (n = 9 follicles), the midfollicular phase (n = 4), or the late follicular phase (after the LH surge, but before ovulation; n = 5). Total RNA extracted from granulosa cells and theca interna at the time of cell isolation or after 24 or 72 h of culture in control or LH-containing medium was subjected to Northern analysis, and autoradiographs were scanned densitometrically. P450scc mRNA levels in granulosa cells were high in the early follicular phase and decreased by 96% after the LH surge (P < 0.05). 3 beta HSD mRNA levels in granulosa cells were 4.2 fold higher in early vs. late follicular phase (P < 0.01). In theca interna, 3 beta HSD mRNA levels were 3.6- and 2.6-fold higher in the early vs. the mid- and late follicular phase (P < 0.05), but levels of P450scc mRNA did not differ significantly with stage of follicular development. After granulosa cells had been cultured for 24 h in control or LH-containing medium, P450scc and 3 beta HSD mRNA had declined dramatically compared to mRNA levels at the time of cell isolation during the early follicular phase (P < 0.01). However, after 72 h in control or LH-containing medium, an increase in P450scc and 3 beta HSD mRNA was observed relative to levels at 24 h (P < 0.01). After 72 h of culture, the signal for P450scc and 3 beta HSD mRNA in granulosa cells exposed to LH was higher than the signal detected in cultures without LH (P < 0.01). Similar changes in message for P450scc were observed in cultured thecal cells. Thus, the previously observed increases in production of progesterone by bovine theca interna and granulosa cells obtained after vs. before the LH surge cannot be explained by an increase in message for P450scc and 3 beta HSD.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8425502 TI - Stress-induced increase in vasopressin and corticotropin-releasing factor expression in hypophysiotrophic paraventricular neurons. AB - The concerted action of CRF and vasopressin (VP) plays a critical role in regulating ACTH release from anterior pituitary cells. In this study, we have explored the expression of these neurohormones in hypophysiotropic paraventricular neurons after repeated exposure of rats to immobilization stress. Cell by cell quantitative in situ hybridization was used to evaluate the steady state level of mRNAs coding for VP and CRF. We found that 16 daily stress exposures resulted in a significant increase in the average cellular level of CRF and VP mRNAs (150% and 200% of control levels, respectively). Moreover, in the repeatedly stressed group, the number of VP-expressing parvicellular neurons was approximately doubled relative to the control value. Using quantitative immunoelectron microscopy, VP- and CRF-immunoreactive sites were assessed in the dense core vesicle compartment of CRF axon terminals in the external zone of the median eminence. We found that after repeated stress, the immunolabeling of VP was augmented, while that of CRF was slightly decreased. Concurrently, we observed a significant increase in the proportion of CRF nerve terminals that were VP positive (from 50% in controls to 90% in stressed animals). We conclude that the observed changes in CRF neurons may represent a physiological response to increased functional demand and may lead to alterations in the composition of the ACTH-releasing signal. PMID- 8425503 TI - Corticosteroid-binding globulin biosynthesis in the mouse liver and kidney during postnatal development. AB - Plasma corticosteroid-binding globulin (CBG) is produced by the liver, but low levels of CBG mRNA have been detected in other tissues, including the kidney. Glucocorticoids influence postnatal renal development in rodents, and CBG production in the kidney may influence the local bioavailability of glucocorticoids. We, therefore, used in situ hybridization and immunohistochemistry to define the sites of CBG biosynthesis during postnatal development and have found that the liver and kidney are major sites of CBG biosynthesis in the first weeks of life. Both CBG and its mRNA were undetectable in the neonatal liver, and only a weak hybridization signal for CBG mRNA was present in the 7-day-old mouse liver. In neonatal mice, the developing tubules of the kidney represent the most active site of CBG biosynthesis, and immunoreactive CBG was also detected in the same cells. By 7 days of age, CBG and its mRNA were colocalized to the proximal convoluted tubules of the juxtamedullary nephrons. The abundance of CBG mRNA in the liver increased from 10 days of age and was accompanied by similar increases in serum CBG until adult levels were reached by 4 weeks of age. In contrast, CBG mRNA in the kidney increased to a maximum during the third week of life, but was undetectable 3 weeks later. The CBG within the proximal convoluted tubules was located in secretory granules close to the luminal surface of the epithelial cells, suggesting that it is secreted into the tubular lumen. Western analysis revealed that marked proteolytic degradation of CBG occurs in the urine concurrently with an increase in CBG biosynthesis in the developing kidney. Thus, the liver is not the only site of CBG biosynthesis in the developing mouse, and CBG production by the epithelial cells of the proximal convoluted tubules may influence glucocorticoid-dependent maturation of the kidney tubules by a process that somehow involves proteolytic degradation. PMID- 8425504 TI - Expression of epidermal growth factor and its receptor in the human ovary during follicular growth and regression. AB - Immunohistochemical studies were performed using specific antibodies to epidermal growth factor (EGF) and EGF receptor to determine their presence and cellular localization in the human ovary during follicular growth and regression. There was no immunostaining for EGF or EGF receptor in primordial follicles. In the preantral follicle stage, immunostaining for EGF and EGF receptor was observed only in the oocyte. The staining intensity of the oocyte increased as the oocyte reached the preovulatory stage. In the antral follicle stage, immunostaining for EGF and EGF receptor became apparent in the granulosa and theca interna cell layers, without appreciable staining in the surrounding stromal cells. The immunostaining for EGF and EGF receptor in the granulosa cells and theca interna cells persisted in preovulatory follicles and corpus luteum, and intensified in the midluteal phase. The stromal cells surrounding the corpus luteum were negative for EGF and EGF receptor staining. In the regressing corpus luteum, immunostaining for EGF and EGF receptor was present in the peripheral lutein cells adjacent to the central core of scar tissue, but absent in the scar tissue of the central core. Corpus albicans showed no staining for EGF and EGF receptor. By contrast, the stromal cells surrounding the corpus albicans in the cortex region demonstrated intense staining for EGF and EGF receptor, while the stromal cells surrounding the corpus albicans in the medullary region were negative for immunostaining. In the case of atretic follicles, the theca interna cells showed intense staining for EGF and EGF receptor, but immunostaining in the scattered granulosa cells was negligible. This is the first study to demonstrate a remarkable change in the expression of EGF and EGF receptor in the oocyte, granulosa cells, thecal cells, and surrounding stromal cells over the course of follicular growth and regression. The results obtained support EGF participation in oocyte maturation and in follicular growth and atresia. The intense immunostaining for EGF and EGF receptor observed in the theca interna cells in atretic follicles and the stromal cells surrounding corpus albicans in the cortex region raises the possibility of EGF involvement in transformation of thecal cells into stromal cells. Furthermore, the cell type-specific simultaneous expression of EGF and EGF receptor in follicular and stromal compartments in the various stages of follicular development suggests that an autocrine mode of EGF action may exist to regulate follicular growth and regression in the human ovary. PMID- 8425505 TI - Distinct promoters in the rat insulin-like growth factor-I (IGF-I) gene are active in CHO cells. AB - In mammals, IGF-I mRNAs contain one of two different leader exon sequences that encode different 5'-untranslated regions (UTRs) and signal peptides. The pattern and regulation of expression of these exon 1 and exon 2-derived mRNAs suggests that the expression of each is controlled by a distinct regulatory region. In order to assess this possibility, DNA fragments consisting of sequences flanking and including the exon 1 and exon 2 transcription initiation sites were cloned into a luciferase expression vector and plasmid DNAs were transiently transfected into Chinese hamster ovary (CHO) cells. A fragment containing approximately 1.1 kb of sequence flanking the most upstream exon 1 transcription initiation site and 362 bp of exon 1 sequence did not stimulate luciferase activity. However, fragments containing 133 bp of 5'-flanking sequence and either 362 or 192 bp of exon 1 sequence stimulated luciferase activity significantly above that seen with a promoterless control plasmid. When the -133/+362 fragment was cloned in the opposite orientation with respect to the luciferase cDNA, the same level of promoter activity was observed. Removal of approximately 860 bp from the inactive fragment (i.e., approximately 782 bp of flanking sequence and approximately 74 bp of exon 1 sequence) resulted in promoter activity which was significantly greater than that seen with the promoterless luciferase expression vector, but which was less than that observed with fragments containing the proximal 133 bp of 5' flanking sequence. Plasmids containing approximately 1.5 kb or 0.5 kb of flanking sequence and 44 bp of exon 2 sequence also significantly stimulated luciferase activity. These results constitute the first demonstration that both exon 1 and exon 2 transcription start sites are associated with distinct and potentially independently regulatable promoters and provide a molecular basis for the differential expression of these leader exons by developmental, tissue-specific and hormonal factors. PMID- 8425506 TI - Alpha-motoneuron excitability at high altitude. AB - It has been hypothesized that chronic hypobaric hypoxia could lead to inhibition of the alpha-motoneuron pool, thus limiting the maximal activation of working skeletal muscles. To test this hypothesis six subjects [32 (SEM 2) years] were evaluated in resting conditions, at sea level and after acclimatization at 5,050 m. The recruitment curves of the Hofmann-reflex (H-) and the direct muscle response. (M-) of the right soleus muscle were obtained by stimulating the posterior tibial nerve with different intensities while recording the electromyogram of the soleus muscle. From the recorded data the net alpha motoneuron excitability (ratio of maximal H-reflex to M-response Hmax:Mmax ratio), the threshold and gain for both responses, obtained from linear regressions through the rising phase of the recruitment curves of both responses, as well as the latency times of both responses were determined. The latency times and the Hmax:Mmax ratio were unchanged at altitude. The thresholds of both responses and the gain of the M-response were unaltered. The gain of the H response was significantly higher at altitude when compared to sea level.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425508 TI - Heat storage and body temperature during cooling and rewarming. AB - During calorimetric experiments with forced cooling and rewarming, changes in rectal temperature (Tre) and mean skin temperature (Tsk) allowed calculations of Burton's (1935) weighting coefficient "a", which relates body temperature change to change in mean body temperature (delta Tb). Calculating delta Tb from change in body heat content (delta Hb), which was determined from direct and indirect calorimetry, included individualized values for body specific heat based on body fat content. In five different cooling procedures there were two with cooling by exposure to cold water and three with cooling in a tubing suit; two of the procedures included mild exercise. The delta Hb ranged from -335 to -1600 kJ; rewarming restored body heat content. The mean (SEM) value of "a" in 119 determinations was 0.75 (0.01). This small variability in the coefficient probably came from the large values of delta Hb and from the use of maximal changes in Tsk and Tre, including afterdrop. Change in Tre by itself correlated with delta Tb, but with much variability. In forced body cooling and rewarming, 0.75 (delta Tre) + 0.25 (delta Tsk) gives an accurate estimate of delta Tb, hence change in body heat storage. PMID- 8425507 TI - Stroke volume response to progressive exercise in athletes engaged in different types of training. AB - Using the impedance cardiography method, heart rate (fc) matched changes on indexed stroke volume (SI) and cardiac output (CI) were compared in subjects engaged in different types of training. The subjects consisted of untrained controls (C), volleyball players (VB) who spent about half of their training time (360 min.week-1) doing anaerobic conditioning exercises and who had a maximal oxygen uptake (VO2max) 41% higher than the controls, and distance runners (D) who spent all their training time (366 min.week-1) doing aerobic conditioning exercises and who had a VO2max 26% higher than VB. The subjects performed progressive submaximal cycle ergometer exercise (10 W.min-1) up to fc of 150 beats.min-1. In group C, SI had increased significantly (P < 0.05) at fc of 90 beats.min-1 (+32%) and maintained this difference up to 110 beats.min-1, only to return to resting values on reaching 130 beats.min-1 with no further changes. In group VB, SI peaked (+54%) at fc of 110 beats.min-1, reaching a value significantly higher than that of group C, but decreased progressively to 22% of the resting value on reaching 150 beats.min-1. In group D, SI peaked at fc of 130 beats.min-1 (+54%), reaching a value significantly higher than that of group VB, and showed no significant reduction with respect to this peak value on reaching 150 beats.min-1. As a consequence, the mean CI increase per fc unit was progressively higher in VB than in C (+46%) and in D than in VB (+105%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425509 TI - Ventilatory response of prepubertal boys and adults to carbon dioxide at rest and during exercise. AB - The aim of this study was to determine whether the greater ventilation in children at rest and during exercise is related to a greater CO2 ventilatory response. The CO2 ventilatory response was measured in nine prepubertal boys [10.3 years (SD 0.1)] and in 10 adults [24.9 years (SD 0.8)] at rest and during moderate exercise (VCO2 = 20 ml.kg-1.min-1) using the CO2-rebreathing method. Three criteria were measured in all subjects to assess the ventilatory response to CO2: the CO2 sensitivity threshold (Th), which was defined as the value of end tidal PCO2 (PETCO2) where the ventilation increased above its steady-state level; the reactivity slope expressed per unit of body mass (SBM), which was the slope of the linear relation between minute ventilation (VE) and PETCO2 above Th; and the slope of the relationship between the quotient of tidal volume (VT) and inspiration time (tI) and PETCO2 (VT.tI-1.PETCO2(-1)) values above Th. The VE, VT, breathing frequency (fR), oxygen uptake (VO2), and CO2 production (VCO2) were also measured before the CO2-rebreathing test. The following results were obtained. First, children had greater ventilation per unit body weight than adults at rest (P < 0.001) and during exercise (P < 0.01). Second, at rest, only VT.tI-1.PETCO2(-1) was greater in children than in adults (P < 0.001). Third, during exercise, children had a higher SBM (P < 0.02) and VT.tI-1.PETCO2(-1) (P < 0.001) while Th was lower (P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425510 TI - Respiratory gas exchange indices used to detect the blood lactate accumulation threshold during an incremental exercise test in young athletes. AB - The time course of changes in blood lactate concentration and ventilatory gas exchange was studied during an incremental exercise test on a cycle ergometer to determine if the lactate accumulation threshold (LT2) could be accurately estimated by the use of respiratory indices (VT2) in young athletes. LT2 was defined as the starting point of accelerated lactate accumulation. VT2 was identified by the second exponential increase in VE and the ventilatory equivalent for O2 uptake with a concomitant nonlinear increase in the ventilatory equivalent for CO2 output. Twelve trained subjects, aged 18-22 years, participated in this study. The initial power setting was 30 W for 3 min with successive increases of 30 W every minute except at the end of the test when the increase was reduced. Ventilatory flow (VE), oxygen uptake (VO2), carbon dioxide output (VCO2), and ventilatory equivalents of O2 and CO2 were determined during the last 30 s of every minute. Venous blood samples were drawn at the end of each stage of effort and analysed enzymatically for lactate concentration. After each test, LT2 and VT2 were determined visually by two investigators from the graphic results using a double-blind procedure. The results [mean (SEM)] indicate no significant difference between LT2 and VT2 expressed as VO2 [43.98 (1.70) vs 44.93 (2.39) ml.min-1 x kg-1], lactataemia [4.01 (0.28) vs 4.44 (0.37) mM.l-1], or heart rate [171 (3.36) vs 173 (3.11) min-1]. In addition, strong correlations were noted between the two methods for VO2 (r = 0.90, P < 0.001), lactataemia (r = 0.75, P < 0.01), and heart rate (r = 0.96, P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425511 TI - Changes in cardiac rhythm in man during underwater submersion and swimming studied by ECG telemetry. AB - A previously reported method for electrocardiographic (ECG) telemetry in water using frequency-modulated current was improved to obtain more stable ECGs. The ECGs of seven healthy men were monitored using the improved method during and after whole-body submersion or underwater swimming. Bradycardia and arrhythmias were observed during the submersion, and transient tachycardia was detected after the start of underwater swimming, followed by bradycardia with arrhythmias. Three different types of arrhythmias were observed: sinus arrhythmia (SA), supraventricular extrasystole (SE) and ventricular extrasystole (VE). SA and SE tended to develop during the latter half of the period of submersion or underwater swimming, and especially after the restart of breathing. VEs were detected in only one subject during submersion, whereas they occurred in most subjects during and after underwater swimming. Individual variations were found in development of arrhythmias, one subject showing no arrhythmia. Bradycardia, SA and SE could depend on vagal suppression in underwater conditions, and VE may be related to the effect of muscular movement on cardiac function in addition to vagal inhibition. PMID- 8425512 TI - Effects of protein supplementation during prolonged exercise at moderate altitude on performance and plasma amino acid pattern. AB - The effects of two levels of protein intake on muscle performance and energy metabolism were studied in humans submitted to repeated daily sessions of prolonged exercise at moderate altitude. For this purpose, 29 healthy males, were exposed to seven successive stages of ski-mountaineering at altitudes between 2500 and 3800 m, and to an isocaloric diet (4000 kcal.day-1, 16,760 kJ.day-1) with either 1.5 g.kg-1.day-1 (C group, n = 14), or 2.5 g.kg-1.day-1 (PR group, n = 15) protein intake. Measurements made after the ski-mountaineering programme did not show any change in body mass. The peak torque during maximal isometric voluntary contraction (MVC) of the quadriceps muscle was unaffected by the repeated exercises, whereas the endurance time at 50% MVC was decreased in PR subjects (-26.8%, P < 0.001). Increased levels of both free fatty acids (+ 147%, P < 0.001) and glycerol (+ 170%, P < 0.001) observed in C subjects would suggest that lipolysis was enhanced after the repeated exercise. The plasma amino acid pattern was altered after completion of the ski-mountaineering programme; the plasma concentration of the three branched-chain amino acids (BCAA) was significantly decreased in C subjects, whereas the higher level of protein intake (PR group) greatly minimized the exercise-induced decrease in serum BCAA. PMID- 8425513 TI - Bone metabolism before and after irradiation with ultraviolet light. AB - The beneficial effects of ultraviolet light on cutaneous vitamin D synthesis, calcium metabolism, and bone formation are well known. Regarding the increasing fear of side effects from ultraviolet B (UV-B), lamps with less energy in the UV B range have been developed. Two spectra with differences in the emission of UV-B have therefore been evaluated for their influence on calcium metabolism. A group of 24 healthy male volunteers was subdivided into two treatment groups. Group 1 was exposed to lamps with higher energy of total UV-B but less energy at the wavelengths below 300 nm than the lamps used in group 2. All subjects were irradiated ten times within 12 days. Exposure time was 3 min in the first session and time of exposure was increased by 10% in every following irradiation (suberythematous doses only). Before the first irradiation, 3 days after the last exposure, and after 4 more weeks, the serum parameters of bone metabolism were determined by standard laboratory methods. Significantly increased levels of 25 hydroxyvitamin D3 were found in both groups. There was only a slight increase of 1,25-dihydroxyvitamin D3. Parathyroid hormone decreased significantly in group 2 only. The data would suggest beneficial effects on bone metabolism for both regimens. The observed effects were more pronounced when shorter wavelengths (group 2) were applied, although the total energy of UV-B was lower in these lamps. PMID- 8425514 TI - Glucose ingestion before and during exercise does not enhance performance of daily repeated endurance exercise. AB - The effect of glucose (Glc) ingestion before and during daily, repeated, prolonged exercise on metabolism and performance was tested. Seven young, healthy males performed cycling exercise in two series, with 1 month interval. Each exercise series consisted of 1 h/day on 3 successive days. On the 3rd day, exercise was continued until exhaustion. The intensity was 73.4 (7.7)% [mean (SD)] of maximal oxygen uptake (VO2max). Glucose (Glc) or placebo (P) drink was ingested 15 min before the start, and at 15 and 45 min of each daily exercise. The total amount of Glc ingested was 43.1 (4.2) g. During exercise, blood Glc concentrations were significantly higher (P < 0.05) when Glc was ingested than when P was ingested [Glc 5.14 (0.32) and P 4.12 (4.17) mmol.l-1 at exhaustion]. However, Glc ingestion did not improve performance time to exhaustion [Glc 92.05 (29.55) and P 98.07 (27.33) min]. Free fatty acid concentrations were significantly lower when Glc was ingested than when P was ingested [Glc 0.63 (0.21) and P 1.39 (0.46) mmol.l-1 at exhaustion]. There were no significant differences in exercise heart rate, VO2, respiratory exchange ratio, blood lactate concentrations or rating of perceived exertion between the conditions nor were there any significant differences in these parameters on different days of exercise. It seems that ingestion of small amounts of Glc does not increase the metabolism of carbohydrate or improve the performance of intensive endurance exercise of poorly trained subjects, even when the exercise is repeated daily. PMID- 8425515 TI - An investigation into the relevance of the pattern of temporal activation with respect to erector spinae muscle endurance. AB - The aim of the present study was to evaluate the viability of a relationship between the temporal activation pattern of parts of the erector spinae muscle and endurance. Seven subjects performed intermittent isometric contractions [4s at 70% maximal voluntary contraction (MVC), 2 s rest] until exhaustion, during which the electromyographical (EMG) activity of the multifidus, iliocostalis thoracis and longissimus muscle segments was recorded. Endurance was defined as the time until exhaustion. Subjects were divided into a high and a low endurance group. The high endurance group showed significantly more variability of EMG amplitude over succeeding contractions. This group demonstrated significantly more alternations of EMG activity between parts of the muscle also. Variability of the EMG amplitude within the contractions did not differ between the groups, nor did MVC. The results indicated that alternating activity between different parts of the erector spinae muscle may function to postpone exhaustion of this muscle as a whole. PMID- 8425516 TI - Electroencephalogram and cardiovascular responses to noise during daytime sleep in shiftworkers. AB - Intermittent noise occurring during sleep has been found to induce heart rate, peripheral vasomotor and electroencephalogram (EEG) changes. This study analysed these responses during the daytime and night-time sleep of shiftworkers doing a three shift system, to determine the influence of the inversion of the sleep-wake cycle on the sensitivity to noise. A group of 14 shiftworkers [aged 37 (SD 5) years] underwent an habituation daytime sleep, two experimental daytime sleeps and a night-time sleep. Traffic noises were presented during sleep [truck, 71 dB(A); motorbike, 67 dB(A); and car, 64 dB(A)] at a rate of nine each hour. The EEG measurements of sleep, electrocardiogram and finger pulse amplitude were recorded continuously. The results were expressed by computing the percentage of observed cardiac response (%HRR) and vasoconstrictive response (%FPR), magnitude of heart rate variation (heart rate response; HRR), percentage of reduction of the digital blood flow (finger pulse response, FPR), cardiac cost (CC = % HRR x HRR) and vasomotor cost (VC = % FPR x FPR). The results showed that, compared to night-time sleep, there was change in the structure of daytime sleep, that is an increase in slow wave sleep (SWS), especially stage 4 sleep decrease of stage 2 and rapid eye movement (REM) sleep latencies, and an earlier SWS and REM sleep barycentric point. During daytime sleep the % FPR was significantly smaller in SWS than in stage 2 or REM sleep. Large differences were observed in % HRR, HRR and CC between daytime sleep stages (SWS less than stage 2 less than REM sleep).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425517 TI - Heredity of muscle fibre composition estimated from a selection experiment in rats. AB - The extent to which muscle fibre composition was determined by the genes transmitted from parents was estimated by using successive selections of rats. A foundation population (G0) was established to carry out the selections; it consisted of 100 albino rats which were randomly chosen from heterogeneous stock. The heterogeneous stock was produced by random-mating of three strains, Wistar Imamichi, Fischer 344 and Donryu. Successive selection for a high percentage of slow twitch fibres (% ST) was made from G0 to the fourth generation (G4). The mean values of % ST changed from 50.0% in G0 to 55.6% in G4. The mean value in G4 was significantly higher than that in G0. The realized heritability in G0-G4 was calculated from the regression of selection response on the cumulated selection differential. The regression coefficient was 0.17 (SEM 0.04). The realized heritability was significantly different from 0 at the 0.05 level. We concluded that in rats about 17% of the variation of muscle fibre composition was determined by the genes transmitted from parents. PMID- 8425518 TI - Does critical swimming velocity represent exercise intensity at maximal lactate steady state? AB - The purpose of this investigation was to determine whether the critical swimming velocity (vcrit), which is employed in competitive swimming, corresponds to the exercise intensity at maximal lactate steady state. vcrit is defined as the swimming velocity which could theoretically be maintained forever without exhaustion and expression as the slope of a regression line between swimming distances covered and the corresponding times. A total of eight swimmers were instructed to swim two different distances (200 m and 400 m) at maximal effort and the time taken to swim each distance was measured. In the present study, vcrit is calculated as the slope of the line connecting the two times required to swim 200 m and 400 m. vcrit determined by this new simple method was correlated significantly with swimming velocity at 4 mmol.l-1 of blood lactate concentration (r = 0.914, P < 0.01) and mean velocity in the 400 m freestyle (r = 0.977, P < 0.01). In the maximal lactate steady-state test, the subjects were instructed to swim 1600 m (4 x 400 m) freestyle at three constant velocities (98%, 100% and 102% of vcrit). At 100% vcrit blood lactate concentration showed a steady-state level of approximately 3.2 mmol.l-1 from the first to the third stage and at 98% of vcrit lactate concentration had a tendency to decrease significantly at the fourth stage. On the other hand, at 102% of vcrit, blood lactate concentration increased progressively and those of the third and fourth stages were significantly higher than those at 100% of vcrit (P < 0.05). These data suggest that vcrit, which can be calculated by performing two timed, maximal effort swimming tests, may correspond to the exercise intensity at maximal lactate steady state. PMID- 8425519 TI - From anemia to cerebellar dysfunction. A review of the ankyrin gene family. AB - The focus of this review is on the ankyrin gene family, key elements in the interaction of the spectrin-based membrane skeleton with the plasma membrane in a variety of tissues and multicellular organisms. The structure/function relationships of ankyrin molecules are reviewed, illustrating how these proteins are uniquely suited to serve as adaptors between the membrane skeleton and a number of integral membrane proteins. Advances in the understanding of ankyrin biology in the brain are discussed and used to show how ankyrins may be involved in the establishment and/or maintenance of specialized plasma membrane domains. Finally, recent research in hematological and neurological disorders are reviewed, suggesting that ankyrins have a role in the development of human disease. PMID- 8425520 TI - Purification and characterization of an aminopeptidase A from Staphylococcus chromogenes and its use for the synthesis of amino-acid derivatives and dipeptides. AB - An aminopeptidase with original specificity was purified 3800-fold to homogeneity from a cellular extract of Staphylococcus chromogenes. The enzyme was specific for acidic amino acids (Asp and Glu) at the N-terminus of peptides and thus can be classified as an aminopeptidase A. However, its specificity was not restricted to acidic amino acids: alpha-hydroxy acids such as L-malic and L-lactic acids were also accepted in position P1. The enzyme had a broad specificity for the residue at position P' 1, accepting all types of amino acids, including Pro, in this position. The optimal conditions for the hydrolysis of Asp-Phe-NH2 were pH 9.5 and 60 degrees C. The enzyme was inhibited by chelating agents and serine protease inhibitors. The activity lost by treatment with chelating agents could be restored by Mn2+ or Zn2+ which also stimulated the native enzyme. This suggests that it is a metalloprotease with a serine residue essential for the activity. The native enzyme had an apparent molecular mass of 430 kDa on gradient gel electrophoresis and subunits of 43 kDa as determined by SDS/PAGE. The enzyme catalyzed the synthesis of peptide and amino acid derivatives such as Asp-Phe-OMe (Aspartame) and malyl-Tyr-OEt from L-Asp and L-malic acid as acyl donors and L Phe-OMe and L-Tyr-OEt as nucleophiles, respectively. The use of the enzyme as a reagent in protease-catalyzed peptide synthesis, N-terminal protection and subsequent deprotection, is described. PMID- 8425521 TI - Effect of side-chain structure on inhibition of yeast fatty-acid synthase by cerulenin analogues. AB - Yeast fatty-acid synthase (FAS) inhibition by cerulenin analogs with varying side chain lengths was compared with that of cerulenin, tetrahydrocerulenin and iodoacetamide. Although inhibition by cerulenin was the highest, the analogs having (E,E)-delta 7,10 double bonds showed high inhibition. This strongly suggests that the (E,E)-delta 7,10 double bonds play an important role in the interaction of the inhibitors with the enzyme. It was suggested that the size of the hydrophobic cavity in the condensing enzyme terminates fatty-acid chain elongation by decreasing inhibition by the C18 analog. Like cerulenin itself, the shortest analog (C6) did not induce malonyl-CoA decarboxylase activity. PMID- 8425522 TI - The reaction mechanism of Ca(2+)-ATPase of sarcoplasmic reticulum. Direct measurement of the Mg.ATP dissociation constant gives similar values in the presence or absence of calcium. AB - Combining rapid filtration and rapid acid quenching, we have directly measured, at pH 7.0 and 5 degrees C, the association and dissociation rate constants of Mg.ATP binding to the sarcoplasmic reticulum (SR) ATPase in the presence of 50 microM calcium and 5 mM MgCl2 (3-4 x 10(6) M-1.s-1 and 9 s-1, respectively). Therefore, we have determined the true affinity for Mg.ATP (Kd = 3 microM) in the presence of calcium, which can not be measured at equilibrium because of spontaneous and fast phosphorylation. At low concentrations, Mg.ATP binding is the rate limiting step in the phosphorylation process, and Mg.ATP dissociation is slower than dephosphorylation. The kinetics of Ca2+ binding measured by rapid filtration are biphasic, reflecting a two-step mechanism, both steps being accelerated by Mg.ATP. Combining rapid filtration and rapid monitoring of the intrinsic fluorescence of SR Ca(2+)-ATPase, we showed that rate constants for calcium binding are always lower than those of Mg.ATP binding to an EGTA incubated enzyme. We measured dissociation and association rate constants of Mg.ATP binding in the absence of calcium (k-1 = 25 s-1 and k1 = 7.5 10(6) M-1.s 1). This gives a Kd similar to that obtained by equilibrium measurements (3-4 microM). Both non-phosphorylated conformations of the enzyme have similar affinity for Mg.ATP. Therefore, activation of ATPase activity by an excess of ATP cannot be explained by a change in affinity of the non-phosphorylated enzyme for Mg.ATP. In conjunction with previous results, these data are used to discuss the molecular mechanism for the Ca(2+)-ATPase cycle, in which ATP is sequentially substrate and activator on a multiple-function single site. PMID- 8425523 TI - Action spectrum of light-harvesting-II precursor apoprotein transcript accumulation and coordinated translation. AB - In an attempt to ascertain that the phytochrome chromoproteins do indeed constitute the photoreceptor involved in light-harvesting-II-apoprotein transcript accumulation and the synchronization of the endogenous circadian clock [Tavladoraki, P., Kloppstech, K. & Argyroudi-Akoyunoglou, J. H. (1989) Plant Physiol. (Bethesda) 90, 665-672], we studied the action spectrum of Cab transcription in 9-10-day-old etiolated Phaseolus vulgaris leaves. Transcript accumulation was detected by hybridization experiments or immunoprecipitation of the in-vitro translation products, obtained from isolated leaf poly(A)-rich mRNA species. It was found that the accumulation of the precursor-light-harvesting-II apoprotein mRNA depends highly on the wavelength of light exposure, the action spectrum of its accumulation being closely correlated with the absorption spectrum of the red-absorbing phytochrome form (Pr); the reversal by far-red light of the light-induced Cab-mRNA accumulation depends also on wavelength of light exposure, used prior to far red for activation of transcription; the action spectrum thus obtained resembles the difference absorption spectrum of the purified phytochrome protein (delta A Pr-Pfr). In addition, immunoblot analysis of total lithium-dodecyl-sulphate-solubilized protein extracts of leaves showed that the appearance of the mature light-harvesting-II apoprotein in etiolated leaves previously exposed to a light pulse of equal dose but varying wavelength, closely follows the pattern of its transcript accumulation and the action spectrum of protochlorophyllide photoconversion. Thus, translation of the protein in vivo, as monitored by the steady level of the protein appearing in Western blots, is closely coordinated with the accumulation of its transcript, the action spectrum of both processes closely resembling the absorption spectrum of phytochrome. The results suggest that phytochrome is involved in photoreception mediating Cab-gene transcription in 9-10-day-old etiolated bean leaves and that Cab transcription, appearance of the mature protein and stabilization of the apoprotein in thylakoids by the chlorophyll formed during the light pulse are in close coordination. PMID- 8425524 TI - Doxorubicin-induced lipid peroxidation and glutathione peroxidase activity in tumor cell lines selected for resistance to doxorubicin. AB - Doxorubicin-induced lipid peroxidation was evaluated in four human or murine cell strains in culture and in their doxorubicin-resistant variants, by the quantification of malondialdehyde produced after a 2-h incubation of cells with the drug. Significantly increased malondialdehyde levels were obtained 24 h after doxorubicin treatment in three of the wild-type cell lines with doses as low as 0.05-0.1 micrograms/ml, which is within an order of magnitude of the concentration of the drug which inhibits cell growth by 50%. This production of malondialdehyde was abolished in two doxorubicin-resistant strains, even with high doses of drug (100-300 micrograms/ml), but was maintained in the third resistant line. No malondialdehyde production was observed in the fourth cell line, sensitive or resistant. It is remarkable that an enhancement of selenium dependent and non-selenium-dependent glutathione peroxidase activities was exhibited during the acquisition of resistance to doxorubicin in the two first lines, but not in the third, whereas a constitutively high non-selenium-dependent glutathione peroxidase activity existed in the doxorubicin-sensitive and doxorubicin-resistant variants of the fourth cell line. Gene expression of selenium-dependent glutathione peroxidase and of glutathione S-transferase pi, which is known partially to bear a non-selenium-dependent glutathione peroxidase activity, were correlated with the corresponding enzyme activities. It appears, therefore, that the already known enhancement of glutathione peroxidase activity and expression in doxorubicin-resistant cell lines has a quantifiable consequence upon doxorubicin-induced lipid peroxidation and may have consequences in the mechanism of resistance to this drug. PMID- 8425525 TI - A purification method and N-glycosylation sites of a 36-cysteine-containing, putative cell/cell adhesion glycoprotein gp64 of the cellular slime mold, Polysphondylium pallidum. AB - A 64-kDa membrane-bound glycoprotein (gp64) of the cellular slime mold Polysphondylium pallidum, is a putative cell/cell adhesion protein identified by adhesion-blocking antibody fragments (Fab). gp64 is expressed on the cell surface of growth-phase cells and seems to mediate cell/cell adhesion. This paper describes an improved purification method based on the lipophilic nature of this protein. A critical step in the purification method is to collect an insoluble top layer appearing during ammonium sulfate precipitation. The sequence of cDNA encoding gp64 and its deduced amino acid sequence have been determined previously. Based on cDNA sequence data, the structure of gp64 protein was analyzed: almost all amino acid compositions and partial amino acid sequences of lysylendopeptidase-digested peptides of gp64 were determined by protein analysis; all six asparagine-linked glycosylation sites (Asn-Xaa-Ser/Thr) in fact contain carbohydrates, and all 36 cysteine residues were involved in forming disulfide bridges. From these data, gp64 seems to be a unique protein among cell/cell adhesion proteins. PMID- 8425526 TI - Characterization of prostaglandin-F2 alpha-binding sites on rat hepatocyte plasma membranes. AB - Prostaglandin (PG) F2 alpha has previously been shown to increase glucose output from perfused livers and isolated hepatocytes, where it stimulated glycogen phosphorylase via an inositol-trisphosphate-dependent signal pathway. In this study, PGF2 alpha binding sites on hepatocyte plasma membranes, that might represent the putative receptor, were characterized. Binding studies could not be performed with intact hepatocytes, because PGF2 alpha accumulated within the cells even at 4 degrees C. The intracellular accumulation was an order of magnitude higher than binding to plasma membranes. Purified hepatocyte plasma membranes had a high-affinity/low-capacity and a low-affinity/high-capacity binding site for PGF2 alpha. The respective binding constants for the high affinity site were Kd = 3 nM and Bmax = 6 fmol/mg membrane protein, and for the low-affinity site Kd = 426 nM and Bmax = 245 fmol/mg membrane protein. Specific PGF2 alpha binding to the low-affinity site, but not to the high-affinity site, could be enhanced most potently by GTP[gamma S] followed by GDP[beta S] and GTP, but not by ATP[gamma S] or GMP. PGF2 alpha competed most potently with [3H]PGF2 alpha for specific binding to hepatocyte plasma membranes, followed by PGD2 and PGE2. Since the low-affinity PGF2 alpha-binding site had a Kd in the concentration range in which PG had previously been shown to be half-maximally active, and since this binding site showed a sensitivity to GTP, it is concluded that it might represent the receptor involved in the PGF2 alpha signal chain in hepatocytes. A biological function of the high-affinity site is currently not known. PMID- 8425527 TI - Purification and characterization of ribonucleoproteins from pea chloroplasts. AB - RNA-binding proteins are known to mediate the post-transcriptional regulation of genes in many organisms. Recently they have been found to be important in the expression of plastid genes. We have purified a group of three single-stranded nucleic-acid-specific acidic proteins (33, 30 and 28 kDa) from chloroplast extracts of pea (Pisum sativum L.), using single-stranded DNA affinity chromatography. All of them have acidic amino termini but the amino acid sequences are unique to each polypeptide, with partial similarities to the recently reported ribonucleoproteins from tobacco chloroplasts. The pea proteins are also antigenically distinct, as shown by Western blot analysis using polyclonal antisera for purified proteins. Further, from their large nucleic-acid binding domains and the polynucleotide substrate affinities, they are predicted to belong to a family of pea plastid ribonucleoproteins. In vivo radiolabeling of proteins in the presence of translational inhibitors as well as in vitro translation of leaf tissue RNA suggest that these proteins are encoded in the nucleus. Antibody cross-reactivity experiments reveal that their genes are conserved during plastid evolution. PMID- 8425528 TI - The use of lac-type promoters in control analysis. AB - For control analysis, it is necessary to modulate the activity of an enzyme around its normal level and measure the changes in steady-state fluxes or concentrations. We describe an improved method for effecting the modulation, as elaborated for Escherichia coli. The chromosomal gene, encoding the enzyme of interest, is put under the control of a lacUV5 or a tacI promoter. The alternative use of the two promoters leads to an expression range which should make it suitable for the use in control analysis of many enzymes. The lacUV5 promoter should be used when the wild-type expression level is low, the tacI promoter when the latter is high. The endogenous lac operon is placed under the control of a second copy of the lacUV5 promoter and a lacY7am mutation (eliminating lactose permease, the transport system for the inducer isopropyl thio-beta-D- galactoside) is introduced. The method was demonstrated experimentally by constructing E. coli strains, in which the chromosomal atp operon is transcribed from the lacUV5 and the tacI promoter. We measured the concentration of the c subunit of H(+)-ATPase, and found that the expression of this enzyme could be modulated between non-detectable levels and up to five times the wild-type level. Thus, in the absence of inducer, no expression of atp genes could be detected when the atp operon was controlled by the lacUV5 promoter, and we estimate that the expression was less than 0.0025 times the wild-type level. We show that the introduction of a lacY mutation facilitated the attainment of steady induction levels of partially induced cells. The mutation also reduced positive cooperativity in the dependence of expression on the concentration of isopropyl-thio-beta-D-galactoside (the inducer) and shifted the concentration of inducer needed for half maximum induction to higher values. These properties should facilitate the experimental modulation of the enzyme activity by varying the concentration of the inducer. PMID- 8425529 TI - Partial cloning of a squid microtubule-associated protein (MAP H1) and the identification of the microtubule binding domain. AB - An approximately 420-kDa ATP binding protein, referred to as MAP H1, has previously been shown to be involved in microtubule-dependent vesicle motility in the squid giant axon. To gain further insight into the structure and function of this protein, partially overlapping cDNA clones encoding approximately a quarter of the MAP H1 molecule were identified from two squid optic lobe libraries using affinity-purified antibodies to squid MAP H1. One clone in particular (KS18), which hybridizes to an approximately 13-kb message, encodes a series of almost identical repeats of a 16-amino-acid sequence that is tandemly repeated. The sequence of clone KS18 is unique and does not correspond to any nucleotide or amino acid sequence in the data base. The presence of repeated elements within the microtubule binding domain of several other MAPs prompted us to investigate whether the MAP H1 repeats are involved in microtubule binding. In-vitro synthesized polypeptides containing these repeats sediment with taxol-stabilized microtubules in a microtubule binding assay. The predicted secondary structure of the 16-amino-acid repeat region of MAP H1 contains alternating beta-sheets and turns and could form the globular domain seen in negative-stain electron micrographs of MAP H1. PMID- 8425530 TI - NMR solution structure of the [Ala26]parathyroid-hormone-related protein(1-34) expressed in humoral hypercalcemia of malignancy. AB - The structure of the biologically active N-terminal domain of human parathyroid hormone-related protein (residues 1-34) containing an Ala substituted for a His in position 26 was studied by two-dimensional proton NMR spectroscopy. Unambiguous NMR assignments of all backbone and side-chain hydrogens were made with the aid of totally correlated spectroscopy experiments, which provided through-bond 1H-1H connectivities, and NOE spectroscopy, which provided through space and sequential backbone connectivities. The NMR data were utilized in distance-geometry algorithms to generate a family of structures. The major structural features include two segments of alpha-helix extending from Glu4 to Lys13 and from Leu27 to Thr33, with two turns from Gln16 to Arg19 and Phe22 to His25. A salt-bridge appears likely between Arg20 and Glu30 which may be critical for holding the receptor-binding domain together. PMID- 8425531 TI - Protein flexibility and aggregation state of human epidermal growth factor. A time-resolved fluorescence study of the native protein and engineered single tryptophan mutants. AB - A time-resolved fluorescence spectroscopic study of the recombinant human epidermal growth factor (hEGF), a bis(tryptophan)-containing protein (Trp49 Trp50), and of the two single-tryptophan-containing engineered mutants with Trp49 or Trp50 replaced by Phe ([W49F]hEGF, [W50F]hEGF), was undertaken in order to gain insight into the conformational dynamics of the C-terminal region. Quite different position-dependent microenvironments for the two Trp residues are shown by comparing the fluorescence intensity decay of both mutants. Trp50 in the single-tryptophan mutant [W49F]EGF probably undergoes a dominant interaction with the solvent. A more heterogeneous environment of Trp49 in the [W50F]hEGF mutant is found. Moreover, the fluorescence decay of the native hEGF is not simply the additive result of the decays of both mutants: the Trp2 sequence confers a conformation of the C-terminal sequence which is more in contact with the rest of the protein molecule. By contrast, the fluorescence anisotropy decay of the native protein is quite similar to that of the single-tryptophan mutants. A high degree of rotational freedom in the C-terminal region of the protein is demonstrated. The resonance energy transfer, which could contribute to the anisotropy decay, appears therefore not to be highly efficient with respect to the depolarization motions. In addition to these local conformational and dynamic aspects of the hEGF C-terminal sequence, the fluorescence anisotropy decay data demonstrate the existence of a dimerization process of the native protein which is dependent on pH and protein concentration. This phenomenon influences the excited-state lifetime profiles and, therefore, the local conformational equilibrium of the C-terminal region. PMID- 8425532 TI - NADH binding site and catalysis of NADH peroxidase. AB - The structure of the complex between cofactor NADH and the enzyme NADH peroxidase from Streptococcus faecalis 10C1 (Enterococcus faecalis) has been determined by crystal soaking, X-ray data collection, model building of NADH and refinement at 0.24-nm resolution based on the known enzyme structure [Stehle, T., Ahmed, S. A., Claiborne, A. & Schulz, G. E. (1991) J. Mol. Biol. 221, 1325-1344]. Apart from NADH, the catalytic center of the enzyme contains FAD and a cysteine that shuttles between thiolate and sulfenic acid states. Unfortunately, this cysteine was irreversibly oxidized to a cysteine sulfonic acid in the established enzyme structure. Based on the geometry of the catalytic center, we discuss the stabilization of the oxidation-sensitive sulfenic acid and propose a reaction mechanism. PMID- 8425533 TI - A two-dimensional NMR study of Co(II)7 rabbit liver metallothionein. AB - The 600-MHz 1H-NMR NOESY spectra on Co(II)7-reconstituted metallothionein (Co7MT), exhibiting hyperfine signals in the range 350 ppm to -50 ppm, with nuclear relaxation times of the order of a few milliseconds, have been measured and several interproton connectivities have been detected. To our knowledge, this is the largest spectral window ever reported for a two-dimensional 1H-NMR spectrum in the case of a paramagnetic metalloprotein. No scalar connectivities could be detected. The hyperfine-shifted signals belong to the cysteine-ligand protons of the Co4S11 cluster of Co7MT. Together with results from one dimensional NOE experiments, the two-dimensional experiments allowed us to proceed with the pairwise assignment of the isotropically shifted signals of the C beta H2 groups of the metal-coordinated cysteines. With the aid of computer graphics inspection of the four-metal-cluster domain, based on the NMR solution structure of Cd7MT, it is possible to purpose sequence-specific assignments of a few hyperfine-shifted 1H-NMR signals. In particular, a tentative assignment is given for the six signals whose shifts exhibit an antiCurie temperature dependence. The assignment relies on the theoretical model that qualitatively rationalizes the isotropic-shift pattern and its temperature dependence. Inferences on the solution structure of the Co4S11 cluster are drawn. PMID- 8425534 TI - Differential insertion of insulin receptor complexes into Triton X-114 bilayer membranes. Evidence for a differential accessibility of the membrane-exposed receptor domain. AB - In the present study, the Triton X-114 phase-separation system has been used to characterize molecular properties of the membrane-exposed domain of an integral membrane hormone receptor. This approach provides novel details of the structure/function relationship of insulin receptors. Upon raising the temperature of a micellar Triton X-114 solution above the cloud-point, a detergent enriched phase pellets and coprecipitates 95% of the purified insulin free (alpha beta)2 receptors. In contrast, 83% of the hormone bound (alpha beta)2 receptor complexes prefer the detergent-depleted phase, exhibiting prominent properties of non-membraneous proteins. Kinetic studies show that, following insulin binding, the amphiphilicity of the receptor complexes is immediately altered. Only monodisperse (alpha beta)2 complexes were detected when receptor/insulin complexes of the detergent-depleted phase were analyzed by detergent-free sucrose density centrifugation in the presence of 10 nM insulin. These results can be explained in the light of the lipid-bilayer-like organization of the precipitating Triton X-114; hormone-induced intramolecular alterations of (alpha beta)2 receptors appear to fundamentally restrict access to the membrane-exposed receptor domain. Basically, different molecular properties are found for alpha beta receptors. Only 67% of the insulin-free receptors coprecipitate with the Triton-X-114-enriched phase; following insulin binding the coprecipitation is only decreased to 42%. In contrast to (alpha beta)2 receptors, formation of noncovalently aggregated receptor complexes, which are detected by sucrose density centrifugation, could account for the exclusion of alpha beta receptor species from Triton X-114 membranes. PMID- 8425535 TI - Human and rodent Alzheimer beta-amyloid peptides acquire distinct conformations in membrane-mimicking solvents. AB - The major constituent of senile plaques (one of the hallmark lesions of Alzheimer's disease) is a 42(43)-amino-acid polypeptide, termed the A4 or beta amyloid peptide. The beta-amyloid peptide or A4 is derived from one or more larger beta-amyloid precursor proteins. The precursor protein from whence the A4 peptide is derived is highly conserved throughout evolution, and humans, monkeys, dogs, and bears develop brain deposits of A4 peptide in amyloid fibrils. However, similar accumulations of A4 amyloid are negligible in the brains of rats and mice for reasons that remain unexplored. Notably, the A4 sequence of rodents, deduced from the cDNA clones, differs only in three amino acids from the A4 isolated from the brain of humans. Hence, these differences could account for the inability of rodents to develop Alzheimer-like A4 amyloid plaques. To test this hypothesis directly, using physical and chemical model systems, we synthesized, purified, and characterized A4 peptides corresponding to the human and rodent sequences. Circular dichroic and Fourier-transform infrared spectroscopy were used with various membrane-mimicking solvents, different peptide concentrations, and variable pH to identify those environmental conditions that promoted beta-pleated sheet formation of the human versus rodent A4. At an intermediate alkaline pH (< or = 10), the rodent peptide has more beta-pleated sheet structure than the human sequence. The beta-pleated sheets for both peptides could be eliminated at very high pH (> or = 12). The amount of the beta-structure increased in an octyl glucoside solution, compared to that found in SDS, as well as in several of the other solutions tested here. This suggests that particles originated from prior membrane damage may play a role in the stabilization of beta-pleated sheets with subsequent formation of amyloid deposits. Finally, we found that higher beta pleated sheet content was observed for the rodent sequences in acetonitrile/water mixtures. In contrast, more beta-pleated sheets were detected with the human A4 in trifluoroethanol/water mixtures at neutral pH. Remarkably, at relatively low peptide concentrations, only the human sequences assumed an extended secondary structure. These data suggest that subtle inter-species amino-acid differences may account for the inability of the rodent peptide to form amyloid fibrils in situ. PMID- 8425536 TI - The cooperative binding of phenylalanine to phenylalanine 4-monooxygenase studied by 1H-NMR paramagnetic relaxation. Changes in water accessibility to the iron at the active site upon substrate binding. AB - The effect of the paramagnetic high-spin Fe(III) ion in phenylalanine 4 monooxygenase (phenylalanine hydroxylase, EC 1.14.16.1) on the water proton longitudinal relaxation rate has been used to study the environment of the iron center. The relaxation rate was measured as a function of the concentration of enzyme, substrate (phenylalanine), inhibitor (noradrenaline) and activator (lysolecithin), as well as of the temperature (18-40 degrees C) and the external magnetic field strength (100-600 MHz). From the frequency dependence of the relaxation rate, an effective correlation time (tau c) of 4.2(+/- 0.5) x 10(-10) s was calculated for the enzyme-substrate complex, which most likely represents the electron spin relaxation rate (tau s) for Fe(III) (S = 5/2) in this complex. The relaxation rate was proportional to the concentration of enzyme (0.04-1 mM) both in the absence and presence of phenylalanine, but the paramagnetic molar relaxivity at 400 MHz and 22 degrees C decreased from 2.2(+/- 0.05) x 10(3) s-1.M 1 in the enzyme as isolated to 1.2(+/- 0.06) x 10(3) s-1.M-1 in the presence of saturating concentrations of the substrate. The activation energy of the relaxation rate also decreased from 11.3 +/- 0.8 kJ/mol to -1.5 +/- 0.2 kJ/mol upon incubation of the enzyme with 5 mM phenylalanine. The results obtained can be interpreted in terms of a slowly exchanging water molecule coordinated to the catalytic paramagnetic Fe(III) in the native and resting enzyme, and that this water molecule seems to be displaced from coordination on the binding of substrate or inhibitor. Moreover, the effect of increasing concentrations of phenylalanine and noradrenaline on the water proton relaxation rate and on the hydrophobic surface properties of the enzyme indicate that substrate and inhibitor induce a similar cooperative conformational change upon binding at the active site. By contrast, the activator lysolecithin does not seem to affect the interaction of water with the catalytic Fe(III). PMID- 8425537 TI - Mutations that significantly change the stability, flexibility and quaternary structure of the l-lactate dehydrogenase from Bacillus megaterium. AB - In order to investigate the physical basis of protein stability, two mutant L lactate dehydrogenases (LDH) and the wild-type enzyme from Bacillus megaterium were analyzed for differences in quaternary structure, global protein conformation, thermal stability, stability against guanidine hydrochloride, and polypeptide chain flexibility. One mutant enzyme, ([T29A, S39A]LDH), differing at two positions in the alpha-B helix, exhibited a 20 degrees C increase in thermostability. Hydrogen/deuterium exchange revealed a rigid structure of this enzyme at room temperature. The substitutions Ala37 to Val and Met40 to Leu destabilize the protein. This is observable in a greater susceptibility to thermal denaturation and in an unusual monomer/dimer/tetramer equilibrium in the absence of fructose 1,6-bisphosphate Fru(1,6)P2. The stability, flexibility and protein-conformation measurements were all performed in the presence of 5 mM Fru(1,6)P2, i.e. under conditions where the three investigated LDH species are stable tetramers. Tryptophan fluorescence was used to monitor the unfolding in guanidine HCl of two local structures in or very close to the beta-sheets at the protein surface. The LDHs form folding intermediates in guanidine HCl that aggregate at elevated temperatures. Pronounced differences between the three investigated enzymes are found in their ability to aggregate. The exchange of Thr29 and Ser39 for Ala leads to significantly less aggregation in guanidine HCl than is observed for wild-type LDH. Using 8-anilinonaphthalene-1-sulfonic acid, the folding intermediates were shown to be in accordance with molten-globule-like structures. We have found, by means of molecular sieve chromatography, that the [T29A, S39A]LDH with its increased thermostability has lower susceptibility to disintegrate into monomers in guanidine HCl at 25 degrees C. Despite the differences in aggregation at low guanidine HCl concentrations and temperatures above 25 degrees C, the molten-globule-like structures of the three investigated LDH species are structurally similar, as shown by molecular-sieve chromatography. Although the thermostabilities of the three LDH species are so different in aqueous buffers, their stabilities in guanidine HCl at 20 degrees C are, surprisingly, almost identical. Some comments are made as to the origin of the observed difference between thermal and guanidine HCl stabilities of the LDH. Near-ultraviolet and far-ultraviolet circular dichroism measurements, as well as differences in the amount of activation by Fru(1,6)P2, point to small global structural rearrangements caused by the mutations. Conformational changes upon Fru(1,6)P2 binding or point mutations in the alpha-B helix show that the Fru(1,6)P2-binding site and the alpha-B helix are structurally linked together.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8425538 TI - Purification, complete amino acid sequence and structural characterization of the heat-stable sweet protein, mabinlin II. AB - A new sweet protein, named mabinlin II, was extracted with 0.5 M NaCl solution from the seeds of Capparis masaikai Levl. and purified by ammonium sulfate fractionation, carboxymethylcellulose-Sepharose ion-exchange chromatography and gel filtration. The sweetness of mabinlin II was unchanged by at least 48 h incubation at nearly boiling temperature. Purified mabinlin II thus obtained gave a single band having a molecular mass of 14 kDa on SDS/PAGE. In the presence of dithiothreitol, mabinlin II gave two bands having molecular masses of 4.6 kDa and 5.2 kDa on SDS/PAGE. Two peptides (A chain and B chain) were separated from reduced and S-carboxamidomethylated mabinlin II by HPLC. The amino acid sequences of the A chain and B chain were determined by the automatic Edman-degradation method. The A chain and B chain consist of 33-amino-acid and 72-amino-acid residues, respectively. The A chain is mostly composed of hydrophilic amino acid residues and the B chain also contains many hydrophilic residues. High similarity was found between the amino acid sequences of mabinlin II and 2S seed storage proteins, especially 2S albumin AT2S3 in Arabidopsis thaliana (mouse-ear cress). PMID- 8425539 TI - The structure of DNA junctions and their interaction with enzymes. PMID- 8425540 TI - Ribonucleases from the extreme thermophilic archaebacterium S. solfataricus. AB - A purification procedure consisting of DEAE-Sephacel chromatography, heparin Sepharose CL-6B chromatography and Mono-S chromatography led to the isolation of three proteins endowed with RNase activity from the thermoacidophilic archaebacterium Sulfolobus solfataricus. They were referred to as p1, p2 and p3, according to their elution order from the Mono-S column. Complete amino acid sequence of p2 and partial sequence of p3 displayed high sequence similarity to the 7-kDa DNA-binding proteins previously isolated in Sulfolobus strains [Choli, T., Wittman-Liebold, B. & Reinhardt, R. (1988) J. Biol. Chem. 263, 7087-7093]. The molecular mass of p2, calculated from sequence data, was 7.02 kDa, which compares fairly well with the value of 7.4 kDa determined by SDS/PAGE. Gel filtration of the molecule under native conditions displayed, however, a largely prevailing form with an assessed molecular mass of 13.0 kDa, which points to a dimeric structure. Kinetic characterization of protein p2 showed a broad pH optimum in the range 6.7-7.6 using yeast RNA as substrate; also, it was shown that activity was unaffected by EDTA, Mg2+ and phosphate. The enzyme did not accept as substrate any homopolyribonucleotide, which points to a rather narrow substrate specificity. This was also confirmed by incubating p2 with tRNA(fMet)Met (fMet, N-formylmethionine) from Escherichia coli: the hydrolysis products were thus identified as 3'-phosphooligonucleotides. PMID- 8425541 TI - Three-dimensional structure of the ternary complex between ribonuclease T1, guanosine 3',5'-bisphosphate and inorganic phosphate at 0.19 nm resolution. AB - The ternary complex formed between RNase T1, guanosine 3',5'-bisphosphate (3',5' pGp) and Pi crystallizes in the cubic space group I23 with a = 8.706(1) nm. In a previous publication [Lenz, A., Heinemann, U., Maslowska, M. & Saenger, W. (1991) Acta Crystallogr. B47, 521-527], the structure of the complex (in which Pi was not located) was described at a resolution of 0.32 nm. This is now extended to 0.19 nm with newly grown, larger crystals. Refinement with restrained least squares converged at R = 17.8% for 8027 reflections with [Fo[ > or = 1 sigma ([Fo[); the final model comprises 120 water molecules. 3',5'-pGp is bound to RNase T1 in the anti form, with guanine in the specific recognition site; the 3' phosphate protrudes into the solvent, and the 5'-phosphate hydrogen bonds with Lys41 O and Asn43 N4. A tetrahedral anion assigned as Pi occupies the catalytic site and hydrogen bonds to the side chains of Tyr38, Glu58, Arg77 and His92. The overall polypeptide fold of RNase T1 in the cubic space group does not differ significantly from that in the orthorhombic space group P2(1)2(1)2(1) except for changes < or = 0.2 nm in loop regions 69-72 and 95-98. PMID- 8425542 TI - cDNA cloning, structural features, and eucaryotic expression of human TAG 1/axonin-1. AB - Axonal surface glycoproteins, composed of repeated immunoglobulin-like and fibronectin-type-III(FNIII)-like domains, mediate adhesion between axons or between axons and non-neuronal cells or extracellular matrix proteins. Several representatives of this group promote neurite outgrowth, when presented as substratum to neurons in culture, and have been implicated in axonal guidance mechanisms. TAG-1 and axonin-1 are presumptive species homologues of the rat and the chick, respectively; together with F11/F3, they form a subgroup of Ig/FNIII like molecules containing a glycosyl-PtdIns membrane anchor. Recent reports on tumor suppressor genes encoding Ig-like and FNIII-like sequences prompted us to isolate the human homologue to TAG-1 and axonin-1. Polymerase chain reaction (PCR) primers were designed to regions conserved in both TAG-1 and axonin-1 using deoxyinosine at ambiguous positions. An expected 1000-bp fragment was obtained from cDNA derived from adult human cerebellum. Using this PCR fragment as a probe, several clones were isolated from a human fetal brain cDNA library. Nucleotide sequence analysis of a full-length clone, as expected, revealed a high degree of similarity to rat TAG-1 (91% identity) and chicken axonin-1 (75% identity) at the amino acid level. The encoded protein was then transiently expressed in monkey COS1 cells, and a stable mouse myeloma cell line was established expressing human TAG-1/axonin-1. The transfected COS1 and myeloma cells showed immunoreactivity on the cell surface with polyclonal anti-(chicken axonin-1) serum. On Western blots, the same antibodies recognized the recombinant protein migrating slightly slower on SDS/PAGE than chicken axonin-1. A comparison of chicken and human brain-tissue proteins by Western-blot analysis revealed a similar apparent molecular mass difference between the two species, which might be due to three additional N-glycosylation sites present on human TAG-1/axonin-1. Immunostaining of cryostat sections of embryonic retinas with polyclonal anti (axonin-1) serum showed similar expression patterns in chicken and human samples at corresponding developmental stages. An additional shared feature of human TAG 1/axonin-1, rat TAG-1 and chick axonin-1 is their attachment to the cell membrane with a glycosyl-PtdIns anchor. PMID- 8425543 TI - Complete amino acid sequence of the regulatory light chain of obliquely striated muscle myosin from earthworm, Lumbricus terrestris. AB - Amino acid sequence analysis of the regulatory light chain of obliquely striated muscle myosin from earthworm, Lumbricus terrestris, was performed completely. The polypeptide consists of 195 amino acids with a calculated molecular mass of 21943 Da. From the arrangement of amino acid residues, the first EF-hand domain appears to be a specific Ca(2+)-binding site. The unusually long N-terminal region of about 40 amino acids, which is characterized by accumulation of basic amino acids, is similar to that of the myosin A1 catalytic light chain from rabbit skeletal muscle. PMID- 8425544 TI - Structural characterization of a biologically active human lipocortin 1 expressed in Escherichia coli. AB - Lipocortin or annexin 1 is a calcium-dependent phospholipid-binding protein which probably acts as a glucocorticoid- regulated anti-inflammatory factor. cDNA for human lipocortin 1 was cloned in the pT7.7 expression plasmid under the control of the inducible bacteriophage T7 RNA polymerase promoter. Upon induction with isopropyl thio-beta-D-galactoside, large amounts of the protein were produced and accumulated in Escherichia coli in a soluble form. The recombinant protein was purified to homogeneity by means of two subsequent ion-exchange chromatographic steps. The final yield was about 30 mg/l bacterial culture. Electrospray mass spectrometric analysis of the purified protein demonstrated that the recombinant product corresponds to the native human lipocortin 1, without the initial methionine and with a free N-terminal alanine; tryptic peptide mapping by fast atom-bombardment mass spectrometry showed that the recombinant protein contains cysteine residues at positions 263 and 324 with free thiol groups, whereas Cys270 and Cys343 are probably involved in an intrachain disulfide bridge. Recombinant human lipocortin 1 reduces the carrageenin-induced paw oedema in rat in vivo and inhibits porcine pancreatic phospholipase A2 activity in vitro; in both cases, a dose-related response is observed. PMID- 8425545 TI - Evidence of serine-protease activity closely associated with Drosophila alcohol dehydrogenase. AB - With the use of monoclonal antibodies against alcohol dehydrogenase (ADH) we detected ADH proteolysis in different Drosophila melanogaster tissues during development [Visa, N., Fiblas, J., Santa-Crus, M. C. & Gonzalez-Duarte R. (1992) J. Histochem. Cytochem. 40, 39-49]. We now report the analysis of this proteolytic activity in crude homogenates and in purified ADH preparations of several Drosophila species. Our results indicate that in non-denaturing IEF gels the proteolytic activity comigrates with native ADH electromorphs of all the species analyzed. In addition, we show that it copurifies with ADH and is responsible for the instability of apparently homogeneous ADH preparations in the presence of SDS. When purified ADH preparations were analyzed, the endogenous proteolytic activity yielded the same banding pattern as that obtained with crude homogenates. Even after rechromatography on Sephacryl S-200, the usual last step in our standard purification protocol, the proteolytic activity remained associated with the ADH fractions. Among the various agents which could explain the ADH-linked proteolytic effect, a pre-existing nicked state of the enzyme or chemical proteolysis have been ruled out. The kinetics observed on pure ADH preparations, the effect of specific protease inhibitors and substrate specificity have led us to ascribe this activity to the subtilase serine-protease family. Given that proteolysis is evident even in rechromatographed Sephacryl S 200 fractions, if incubated in SDS for enough time, we propose two alternative hypotheses to explain this phenomenon. First, the proteolytic activity may come from a protease which is inseparable from the ADH active forms and second, the ADH itself may behave as a subtilase when it adopts a particular conformation. Moreover, the previously reported differential banding pattern during development suggests a role for this activity in vivo, in which fatty acids could produce the inducer effect attributed to SDS in vitro. PMID- 8425546 TI - Fast myosin heavy chain diversity in skeletal muscles of the rabbit: heavy chain IId, not IIb predominates. AB - The myosin heavy chain (HC) composition of various rabbit muscles was analysed at both the mRNA and the protein level. S1-nuclease mapping was performed with a cDNA probe specific for myosin HCIIa, yielding a fully protected sequence for HCIIa, a partially protected sequence for HCIIb, and an additional signal putatively assigned to HCIId. At the protein level, three fast myosin HC isoforms, HCIIa, HCIIb and HCIId, were separated by gradient PAGE. The results obtained at the protein level were in agreement with the findings at the mRNA level. The expression of appreciable amounts of myosin HCIIb, the predominating isoform of fast-twitch muscles in rat and mouse, was restricted in the rabbit to only a few muscles, i.e. adductor magnus, gastrocnemius, latissimus dorsi and vastus lateralis. Typical fast-twitch muscles such as extensor digitorum longus, tibialis anterior and psoas contained only minute amounts of HCIIb. The HCIId isoform, demonstrated in the present study for the first time in rabbit, is the predominating fast myosin HC isoform in this species. Electrophoretic analyses of myosin HC in histochemically defined single fibers confirmed the lack of fibers expressing only HCIIb in tibialis anterior, whereas such fibers were found in the adductor magnus. In addition to fiber types IIB, IID, and IIA expressing HCIIb, HCIId, and HCIIa, respectively, an appreciable amount of hybrid fibers coexpressing two HC isoforms at various ratios were found: HCIIb > HCIId; HCIId > HCIIb; HCIId > HCIIa; HCIIa > HCIId; HCIIa > HCI; HCI > HCIIa. This fiber-type spectrum indicates possible fiber-type transitions in the order IIB<==> IIB<==>IIDB<==>IID<==>IIDA<==>IIAD<==>IIA<==>IIC<==>IC <==>I. PMID- 8425547 TI - Flavin dynamics in reduced flavodoxins. A time-resolved polarized fluorescence study. AB - The time-resolved fluorescence and fluorescence anisotropy characteristics of reduced flavin mononucleotide in solution as well as bound in flavodoxins isolated from the bacteria Desulfovibrio gigas, Desulfovibrio vulgaris, Clostridium beijerinckii MP and Megasphaera elsdenii have been examined. All fluorescence and fluorescence anisotropy decays were analyzed by two different methods: (a) least-squares fitting with a sum of exponentials and (b) the maximum entropy method to yield distributed lifetimes and correlation times. The results of both approaches are in excellent agreement. The fluorescence decay of the free as well as protein-bound reduced flavin chromophore is made up of three components. The shortest component proves to be relatively sensitive to the environment and can therefore be used as a diagnostic tool to probe the microenvironment of the reduced isoalloxazine ring system. The other two longer fluorescence lifetime components are insensitive to the chromophore environment and seem therefore to be related to intrinsic, photophysical properties of the reduced chromophore. Fluorescence anisotropy decays show that the flavin mononucleotide in all four reduced flavodoxins is immobilized within the protein matrix, as indicated by the recovery of a single rotational correlation time, reflecting the rotation of the whole protein. No indications are found that rapid structural fluctuations occur in reduced flavodoxins, and the mechanism of electron transfer from flavodoxin to other redox proteins seems to involve immobilized reduced flavin. PMID- 8425548 TI - Homology of pyridoxal-5'-phosphate-dependent aminotransferases with the cobC (cobalamin synthesis), nifS (nitrogen fixation), pabC (p-aminobenzoate synthesis) and malY (abolishing endogenous induction of the maltose system) gene products. AB - Bacterial deletion mutants have indicated that the gene products of cobC, nifS, pabC and malY participate in important metabolic pathways, i.e. cobalamin synthesis, nitrogen fixation, synthesis of p-aminobenzoate and the regulation of the maltose system, respectively. However, the proteins themselves and their specific functions have not yet been identified. In the course of our studies on the evolutionary relationships among aminotransferases, we have found that the above gene products are homologous to aminotransferases. Profile analysis [Gribskov, M., Luthy, R. & Eisenberg, D. (1990) Methods Enzymol. 183, 146-159] based on the amino acid sequences of certain subgroups of aminotransferases as probes attributed significant Z scores in the range 5-20 SD to the deduced amino acid sequences of the above gene products as included in the protein data base. Reciprocal profile analyses confirmed the homologies. All known aminotransferases are pyridoxal-5'-phosphate-dependent enzymes and catalyze the reversible transfer of amino groups from amino acids to oxo acids. The sequence homologies suggest that the above gene products are aminotransferases or other closely related pyridoxal-5'-phosphate-dependent enzymes probably catalyzing transformations of amino acids involving cleavage of a bond at C alpha. PMID- 8425549 TI - Chemical assay for cyst(e)ine-rich peptides detects a novel intestinal peptide ZF 1, homologous to a single zinc-finger motif. AB - Cysteine is a relatively infrequent constituent of proteins, which in its thiol or half-cystine form contributes in a special manner to their three-dimensional structure. We show that in small cystine-containing peptides, the Cys content is always higher than the average in proteins in general. This observation makes it possible to search for new peptides by monitoring only their Cys content. We have developed a chemical assay for the detection of cyst(e)ine-rich peptides in tissue extracts. Using this assay we have isolated from porcine intestine a novel cysteine-rich peptide, which we denote ZF-1. ZF-1 is homologous to a single zinc finger motif and has an acetylated N-terminus. This is the first demonstration of the existence of a processed single zinc-finger-like structure. The structural homology of ZF-1 to the zinc-finger motif, present in several metal-binding and DNA-binding proteins, suggests an important role of this peptide in metal transport and/or modulation of gene expression. PMID- 8425550 TI - Comparison of the dynamic structure of alpha-chymotrypsin in aqueous solution and in reversed micelles by fluorescent active-site probing. AB - A highly fluorescent anthraniloyl (Ant) group was covalently attached to the active site of alpha-chymotrypsin (CT), probably at Ser195. Ant-CT is stable at neutral pH for months, allowing a detailed fluorescence study of Ant-CT as a model protein to investigate its physical properties in 0.1 M Tris/HCl, pH 8.2, and in reversed micelles of n-octane, 0.1 M Tris/HCl, pH 8.2, and sodium bis(2 ethylhexyl)sulfosuccinate (AOT). Steady-state fluorescence measurements of the progressive red-shift of the center of gravity of the emission band as function of degree of hydration, wo, defined as [H2O]/[AOT], indicate that the average polarity in the vicinity of the probe is approaching that of bulk water at wo > 12. Time-resolved fluorescence measurements of Ant-CT in water and in reversed micelles showed that the active site has different properties in reversed micelles compared to those in water. Some specific changes at very low water content (0.6 < wo < 5) can be observed, which correlate with enzyme activity measurements in the same wo region (unpublished results). These effects are, for instance, significant changes in the average fluorescence lifetime and the internal flexibility of the probe. The overall rotational-correlation time of the enzyme in AOT reversed micelles seems to be independent on wo (5 < wo < 29), which suggests that the enzyme creates its own micelle. PMID- 8425551 TI - Snake-venom phospholipase A2 neurotoxins. Potentiation of a single-chain neurotoxin by the chaperon subunit of a two-component neurotoxin. AB - The venoms from Crotalinae and Viperinae snakes contain only two kinds of phospholipase A2 neurotoxins (beta-neurotoxins): single-chain beta-neurotoxins, such as agkistrodotoxin and ammodytoxin-A, and dimeric beta-neurotoxins, which, in the case of the best studied ones, crotoxin-like toxins, consist of the non covalent association of a phospholipase A2 (CB) and a non-enzymatic chaperon (CA). Possible evolutionary relationships of these beta-neurotoxins have been investigated by analyzing whether CA could behave as a chaperon toward agkistrodotoxin and ammodytoxin, as it does in the crotoxin complex. CA increased the lethal potency of agkistrodotoxin and modified its pharmacological effect on Torpedo synaptosomes. Sedimentation experiments proved that CA can form an heterocomplex with agkistrodotoxin. Agkistrodotoxin prevented the binding to CA of an anti-CA mAb which recognizes an epitope at the zone of interaction between crotoxin subunits, suggesting the association of CA and agkistrodotoxin implicated the same zone. A 10-fold molar excess of CA over ammodytoxin modified the effect of ammodytoxin on acetylcholine release but did not increase the lethal potency of ammodytoxin. Sedimentation experiments showed CA and ammodytoxin can form an heterocomplex which is less stable than CA.agkistrodotoxin. Ammodytoxin A did not compete with the anti-CA mAb. These observations are in good agreement with the sequence similarities between CB and agkistrodotoxin (80%) and ammodytoxin A (60%). PMID- 8425552 TI - Engineered yeast cells as model to study coupling between human xenobiotic metabolizing enzymes. Simulation of the two first steps of benzo[a]pyrene activation. AB - Human microsomal epoxide hydrolase and cytochrome P450 (P450) 1A1 were coexpressed in Saccharomyces cerevisiae from expression cassettes integrated respectively into the host chromosomal DNA and on a multicopy plasmid in a strain already overexpressing yeast NADPH-cytochrome P450 reductase (P450 reductase). A styrene-oxide-hydrolase activity (2 nmol.min-1.mg microsomal protein-1) and a 7 ethoxyresorufin-O-deethylase activity (320 pmol.min-1.mg microsomal protein-1) characteristic respectively of microsomal epoxide hydrolase and P450 1A1 were detected. The conversion of benzo[a]pyrene (B[a]P) to B[a]P-7,8-dihydrodiol both in microsomal preparations and in growing yeast cells was observed, demonstrating an efficient coupling between the two human enzymes. Kinetic analysis indicated that the B[a]P-7,8-oxide produced by the P450-1A1-dependent reaction does not accumulate before hydrolysis by microsomal epoxide hydrolase. This system was also used as a control to evaluate the coupling efficiency of a mixture of microsomes or of yeast cells containing separately the individual enzymes (i.e., human P450 1A1 and microsomal epoxide hydrolase). B[a]P-7,8-oxide was well converted to the corresponding dihydrodiol with a mixture of microsomes. In contrast, when the same experiment was repeated with a mixture of cells expressing independently the two activities, dihydrodiol formation was not observed. Coexpression of human phase I and phase II enzymes in a single yeast cell and microsome mixture thus appear to be complementary tools for the simulation of human-drug-metabolism or carcinogen-metabolism pathways. PMID- 8425553 TI - The Ets family of transcription factors. AB - Interest in the Ets proteins has grown enormously over the last decade. The v-ets oncogene was originally discovered as part of a fusion protein expressed by a transforming retrovirus (avian E26), and later shown to be transduced from a cellular gene. About 30 related proteins have now been found in species ranging from flies to humans, that resemble the vEts protein in the so-called 'ets domain'. The ets domain has been shown to be a DNA-binding domain, that specifically interacts with sequences containing the common core trinucleotide GGA. Furthermore, it is involved in protein-protein interactions with co-factors that help determine its biological activity. Many of the Ets-related proteins have been shown to be transcription activators, like other nuclear oncoproteins and anti-oncoproteins (Jun, Fos, Myb, Myc, Rel, p53, etc.). However, Ets-like proteins may have other functions, such as in DNA replication and a general role in transcription activation. Ets proteins have been implicated in regulation of gene expression during a variety of biological processes, including growth control, transformation, T-cell activation, and developmental programs in many organisms. Signals regulating cell growth are transmitted from outside the cell to the nucleus by growth factors and their receptors. G-proteins, kinases and transcription factors. We will discuss how several Ets-related proteins fit into this scheme, and how their activity is regulated both post- and pre translationally. Loss of normal control is often associated with conversion to an oncoprotein. vEts has been shown to have different properties from its progenitor, which might explain how it has become oncogenic. Oncogene-related products have been implicated in the control of various developmental processes. Evidence is accumulating for a role for Ets family members in Drosophila development, Xenopus oocyte maturation, lymphocyte differentiation, and viral infectious cycles. An ultimate hope in studying transformation by oncoproteins is to understand how cells become cancerous in humans, which would lead to more effective treatments. vEts induces erythroblastosis in chicken. Cellular Ets family proteins can be activated by proviral insertion in mice and, most interestingly, by chromosome translocation in humans. We are at the beginning of understanding the multiple facets of regulation of Ets activity. Future work on the Ets family promises to provide important insights into both normal control of growth and differentiation, and deregulation in illness. PMID- 8425554 TI - Enzyme inactivation and protection during entrapment in reversed micelles. AB - Lactate dehydrogenase (LDH) was found to lose rapidly its catalytic activity during the process of entrapment in hydrated reversed micelles of sodium bis(2 ethylhexyl)sulfosuccinate (AOT) in octane. It was demonstrated that this inactivation was caused by the surfactant which penetrated into the injected enzyme-containing aqueous bulk phase during the short time of mechanical stirring needed to convert the initial biphasic mixture into a monophasic reversed micellar solution. The unfavorable inactivation phenomenon could be efficiently eliminated by the addition of either NADH or pyruvate into the enzyme stock solution prior to its injection into AOT solution in octane. The catalytic activity of substrate-protected LDH in AOT reversed micelles increased with increasing water content of the micellar system, reaching the maximal level above wo = 30 when aqueous inner cores of reversed micelles grew large enough to allow unrestricted accommodation of the enzyme molecule. It is suggested that the employment of substrate-protected enzymes could represent a generally useful approach for producing highly efficient enzyme-containing reversed micellar reaction systems. PMID- 8425555 TI - Rat kidney endopeptidase 24.16. Purification, physico-chemical characteristics and differential specificity towards opiates, tachykinins and neurotensin-related peptides. AB - Endopeptidase 24.16 was purified from rat kidney homogenate on the basis of its ability to generate the biologically inactive degradation products neurotensin (1 10) and neurotensin (11-13). On SDS gels of the proteins pooled after the last purification step, the enzyme appeared homogeneous and behaved as a 70-kDa monomer. The peptidase was not sensitive to specific inhibitors of aminopeptidases, pyroglutamyl aminopeptidase I, endopeptidase 24.11, endopeptidase 24.15, proline endopeptidase and angiotensin-converting enzyme but was potently inhibited by several metal chelators such as o-phenanthroline and EDTA and was blocked by divalent cations. The specificity of endopeptidase 24.16 towards peptides of the tachykinin, opioid and neurotensin families was examined by competition experiments of tritiated neurotensin hydrolysis as well as HPLC analysis. These results indicated that endopeptidase 24.16 could discriminate between peptides belonging to the same family. Neurotensin, Lys8-Asn9 neurotensin(8-13) and xenopsin were efficiently hydrolysed while neuromedin N and kinetensin underwent little if any proteolysis by the peptidase. Analogously, substance P and dynorphins (1-7) and (1-8) were readily proteolysed by endopeptidase 24.16 while neurokinin A, amphibian tachykinins and leucine or methionine enkephalins totally resisted degradation. By Triton X-114 phase separation, 15-20% of endopeptidase 24.16 partitioned in the detergent phase, indicating that renal endopeptidase 24.16 might exist in a genuine membrane-bound form. The equipotent solubilization of the enzyme by seven detergents of various critical miscellar concentrations confirmed the occurrence of a membrane-bound counterpart of endopeptidase 24.16. Furthermore, the absence of release elicited by phosphatidylinositol-specific phospholipase C suggested that the enzyme was not attached by a glycosyl-phosphatidylinositol anchor in the membrane of renal microvilli. Finally, endopeptidase 24.16 could not be released from these membranes upon trypsinolysis. PMID- 8425556 TI - Interaction of staphylokinase with different molecular forms of plasminogen. AB - In order to obtain more information on the mechanism of plasminogen activation by staphylokinase (STA), we have studied the interaction between recombinant STA (STAR) and different molecular forms of human plasminogen, including Glu plasminogen (native moiety), Lys-plasminogen (partially degraded moiety) and low molecular-mass (LMM) plasminogen (moiety lacking kringles 1-4). Addition of 2 microM STAR to 0.4 microM Glu-plasminogen, Lys-plasminogen or LMM plasminogen resulted in the generation of proteolytic activity towards the chromogenic substrate D-Val-Leu-Lys-NH-PhNO2 (S-2251) corresponding to the exposure of 1 active center/plasminogen molecule. Complex formation was associated with conversion of the one-chain plasminogen moieties to two-chain plasmin, and with quantitative conversion of Glu-plasminogen to Lys-plasmin. The stoichiometry of the plasminogen-STAR complex, determined by binding of the complex to Lys Sepharose and measurement of residual STAR, was found to be equimolar. The plasminogen-STAR complexes were inhibited by alpha 2-antiplasmin with second order rate constants of 2.4 +/- 0.17 x 10(6) M-1 s-1 for Glu-plasminogen, 2.4 +/- 0.21 x 10(6) M-1 s-1 for Lys-plasminogen and 9.4 +/- 1.5 x 10(4) M-1 s-1 for LMM plasminogen. Glu-plasmin-STAR, Lys-plasmin-STAR and LMM plasmin-STAR had comparable catalytic efficiencies (kcat/Km) for the activation of Glu-plasminogen (0.24-0.29 microM-1 s-1), Lys-plasminogen (0.57-0.79 microM-1 s-1) or LMM plasminogen (0.11-0.16 microM-1 s-1). In a human plasma milieu in vitro STAR, Glu plasmin-STAR, Lys-plasmin-STAR and LMM-plasmin-STAR were equally effective for the lysis of 125I-fibrin-labeled human plasma clots [50% clot lysis in 2 h (EC50) with 11-13 nM test compound] and equally fibrin-selective (residual fibrinogen levels of 72-84% after 2 h at EC50). Our results thus confirm that plasminogen and STAR form a 1:1 stoichiometric complex in which plasminogen is converted to plasmin and Glu-plasminogen to Lys-plasmin. The lysine-binding sites in kringles 1-4 of plasminogen are not required for the complex formation with STAR, nor for the enzyme activity of the complex with STAR in purified systems and in a human plasma milieu. The lysine-binding sites are, however, important for the rate of the inhibition of the complexes by alpha 2-antiplasmin. PMID- 8425557 TI - Too much of a good thing. "Reductionism run amok". PMID- 8425558 TI - The first official meeting of the International Society for Experimental Hematology. AB - In 1972, 85 scientists from 10 countries came to Milwaukee to participate in the first official meeting of the International Society for Experimental Hematology. We summarize here some of the events leading up to the meeting, fondly recollect the enthusiastic spirit of the meeting and reprint the program, which is not all that different in content from the program of the 21st meeting. PMID- 8425559 TI - Primitive hemopoietic stem cells: direct assay of most productive populations by competitive repopulation with simple binomial, correlation and covariance calculations. AB - Quantitative analyses of primitive hemopoietic stem cell (PHSC) populations are important both for basic biology and for clinical applications. Unfortunately, many conventional assays fail to measure long-term repopulating ability and maximal differentiating ability, the most important characteristics of the PHSC. The competitive repopulation assay described here focuses on this characteristic, assaying the precursors from which most differentiated cells are descended over large fractions of the life span in laboratory mice. Thus long-term repopulating ability and the ability to differentiate into both myeloid and lymphoid lineages are measured directly from 2.5 to 12.5 months after transplantation. This technique also has found high correlations between granulocytes, macrophages, and T and B lymphocytes as early as 3 weeks after transplantation. All or most differentiated cells of these widely disparate types appear to be descended from a common precursor cell, while myeloid-specific or lymphoid-specific precursors produce few or no descendants. However, large increases in variances between 3 and 6 weeks and 12 weeks after transplantation suggest that most of the initially active multilineage precursors are exhausted. Thus the ability to differentiate into widely disparate lineages does not establish long-term repopulating ability. PMID- 8425560 TI - Measurement of serum levels of macrophage colony-stimulating factor (M-CSF) in patients with uremia. AB - Serum levels of monokines, including macrophage colony-stimulating factor (M CSF), tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha) and IL-1 beta (IL-1 beta), were measured in patients with chronic renal failure in an attempt to clarify the kinetics of these cytokines in the course of renal anemia. M-CSF was the only monokine detectable in the serum from all patients as well as healthy donors, making this cytokine feasible and reliable for serial evaluations. On all occasions, the level of M-CSF in uremic patients was significantly higher than that in healthy donors (29.4 +/- 12.3 vs. 5.5 +/- 1.1 ng/mL). In patients undergoing hemodialysis, the serum level of M-CSF was greater than that in patients undergoing continuous ambulatory peritoneal dialysis or in uremic patients without dialysis therapy. No difference was observed, however, in the levels of IL-1 alpha, IL-1 beta and TNF-alpha levels in these groups. Patients with severe anemia were subsequently treated with 60 to 80 U/kg per week of human recombinant erythropoietin (rhEpo) for 3 months. After this replacement therapy, hemoglobin levels increased with a variable change ranging from 0 to 3.5 g/dL. The pretherapy M-CSF level, however, was found to predict statistically the response to the therapy (p < 0.05). Patients with a lower pretherapy value responded better to rhEpo therapy; those with a higher level showed a minor degree of response. From these results, we postulate that the elevated M-CSF serum level in uremic patients is in part a consequence of the dialysis procedure and that rhEpo therapy is more effective in patients who are under sophisticated dialysis protocol and have a lower M-CSF level. PMID- 8425561 TI - Modulation of lymphohematopoiesis in long-term cultures by gamma interferon: direct and indirect action on lymphoid and stromal cells. AB - Gamma interferon (IFN-gamma) is the product of multiple cell types within the bone marrow microenvironment and has been demonstrated to act as a potent inhibitor of myelopoiesis in vitro and in vivo. The action of this cytokine on lymphohematopoiesis has now been examined on both long-term bone marrow cultures and representative cloned cellular components of the bone marrow microenvironment. In myelopoietic (Dexter) cultures, the half maximal inhibitory concentration of IFN-gamma was between 1 and 10 U/mL. In comparable lymphopoietic (Whitlock/Witte) cultures, IFN-gamma inhibited the production of B-lineage lymphoid cells with a half maximal effective concentration of less than 1 U/mL. In a clonal assay for pre-B cells, IFN-gamma inhibited colony formation with a half maximal concentration of 1 to 5 U/mL. Not all B-lineage lymphoid cells displayed the same sensitivity, however. Growth of the IL-7-dependent B cell line (2E8) in methylcellulose assays was unaffected by IFN-gamma while the replication of other lymphoid lines was partially or completely inhibited. IFN-gamma induced the expression of cell surface proteins (MHC Class I and II) on both B-lineage cells and stromal cells. In cloned stromal cell lines, IFN-gamma increased the steady state mRNA levels for the cytokines interleukin-6 (IL-6) and JE, a member of the IL-8 family. These data indicate that IFN-gamma acts within the lymphohematopoietic microenvironment through both direct and indirect actions on the hemopoietic and stromal cell populations. PMID- 8425562 TI - Hematopoietic stem cells engraft in untreated transplant recipients. AB - We investigated the engraftment of hematopoietic stem cells in completely untreated transplant recipients to further study hematopoietic cell regulation and for possible inclusion in gene therapy protocols. Untreated female Balb/c recipients received a single infusion of male Balb/c marrow cells. Donor origin of the hematopoietic cells was determined by polymerase chain reaction (PCR), Southern and in situ hybridization analyses with Y-chromosome-specific probes. We found that up to 47% day 12 CFU-S (26.2 +/- 12.6%, mean +/- SD, range 13.3 to 46.7%), 7.3 +/- 5.5% CFU-GM and from 2.5 to approximately 10% nucleated marrow cells were of donor-origin at 8 weeks after marrow infusion. Our results indicate that hematopoietic stem cells can stably engraft in completely unconditioned recipients but, during the interval analyzed, have a low tendency to differentiate. Moreover, the data suggest that under steady-state conditions, niches for primitive hematopoietic cells present in the marrow microenvironment are not saturated, and are readily available. We conclude that the untreated recipient transplant model, in conjunction with sensitive techniques for the detection of donor cells, provides a valuable means for studying hematopoietic stem cell regulation and indicates a need to reassess our understanding of the interactions between stem cells and the hematopoietic microenvironment. PMID- 8425563 TI - Hematopoiesis on cellulose ester membranes. XIII. A combination of cloned stromal cells is needed to establish a hematopoietic microenvironment supportive of trilineal hematopoiesis. AB - A mixture of stromal cells from murine bone marrow placed upon cellulose ester membranes (CEM) and then implanted intraperitoneally (i.p.) in mice results in a regenerated hematopoietic microenvironment which supports trilineal hematopoiesis. We used this model to study the capacity of 5 cloned murine stromal cell lines of marrow origin to support hematopoiesis in vivo: MBA-1 (fibroblast); MBA-2 (endothelial); MBA-13 (fibroendothelial); 14F1.1 (endothelial adipose); and 14M1.4 (macrophage).10(7) stromal cells of a single cell line were applied to 1.5 cm2 CEM, which were folded into tubes and implanted i.p. into mice. Similarly, combinations of 4, 3 and 2 stromal cell lines were applied to CEM and implanted i.p. Single lines were implanted into syngeneic hosts of the same murine strain from which the clone was derived and into nude mice. Combinations of stromal cells were implanted only in nude mice to avoid allogeneic incompatibility. CEM implants were removed after intervals of 5 to 36 weeks and examined histologically. 1) Stromal cells of a single phenotype did not develop hematopoiesis. 2) A combination of 4 stromal phenotypes (MBA-1, MBA-2, MBA-13 and 14F1.1) formed a hematopoietic microenvironment supportive of trilineal hematopoiesis and bone. 3) The combination of 14F1.1 (endothelial adipose) + a second stromal phenotype--MBA-1 (fibroblast) or MBA-2 (endothelial) or MBA-13 (fibroendothelial) also supported trilineal hematopoiesis and bone. 4) CEM coated with MBA-13 or MBA-1 developed bone but no hematopoiesis. The endothelial-adipose phenotype appears to be essential to support hematopoiesis but requires other types of stromal cells--fibroblast, fibroendothelial or endothelial phenotype. PMID- 8425564 TI - Potentiation of myeloid colony-formation in bone marrow of intact and neonatally thymectomized mice by the thymic hormone THF-gamma 2. AB - The effect of the thymic hormone THF-gamma 2 on committed stem cells of bone marrow (BM) origin was determined using the myeloid progenitor cell clonal assay. Preincubation of normal BM cells with THF-gamma 2 for 1 hour or 18 hours caused a 2- to 6-fold increase in the number of myeloid colonies in the presence of suboptimal concentrations of colony-stimulating factor (CSF). The optimal dose of THF-gamma 2 causing this enhancement was in the range of 25 to 100 ng/mL. THF gamma 2 was not able to replace CSF as an inducer in these experiments. THF-gamma 2 neither induced IL-6 activity upon 24-hour incubation with bone marrow cells nor enhanced LPS-induced IL-6 secretion by bone marrow cells in vitro. Neonatal thymectomy (NTx) of Balb/c mice caused a decrease in myeloid progenitors, which was repaired by serial injections of THF-gamma 2. The repair of the stem cell compartment in the bone marrow correlated with an increased percentage of Thy1+ cells in the spleen of THF-gamma 2-treated NTx mice. These findings indicate that THF-gamma 2 is able to regulate committed stem cell functions in the bone marrow of immune-deprived NTx and of normal mice. PMID- 8425565 TI - A cytoadhesion assay for the binding of cloned hemopoietic progenitor cells to stroma. AB - Cell adhesion molecules responsible for the specific recognition and adhesion that necessarily occur between hemopoietic progenitor cells (HPC) and stromal cells within the bone marrow are likely of multiple nature in the cell membrane. Systems less complex than intact bone marrow, in which the interactions between adhesion molecules and their ligands may be studied, is greatly needed. Using 4 cloned murine IL-3-dependent HPC lines, B6Sut, FDCP-1, FDCP-2 and FDCP-Mix, a system of co-culture has been established and standardized with 2 murine stromal cell lines, GB1/6 and 3T3. HPC were radiolabeled with 51Cr, and an optimal set of conditions was established for the adherence of HPC to stromal cells. It was found that a source of IL-3, whether supplied as recombinant murine IL-3 or WEHI conditioned medium, was a necessary component of the labeling and assay medium to achieve maximal adherence to stroma. Likewise, the presence of serum also resulted in overall better cytoadhesion than did serum-free conditions. All 4 cell lines bound GB1/6 in a reproducible manner from approximately 63% for FDCP-1 to 20% for FDCP-Mix; binding to 3T3 was higher than to GB1/6 for all HPC. Approximately 25 to 30% of the cell populations were not able to adhere to stroma, indicating a fairly constant degree of heterogeneity with respect to expression of adhesion molecules. Cytoadhesion was found to have at least one component that was temperature-sensitive, as adhesion of FDCP-1 to GB1/6 was 41.5 +/- 1.3% at 4 degrees C compared with 63.2 +/- 1.1% at 37 degrees C. The adhesion reaction itself occurred independently of metabolic energy production and relied on the presence of receptor-ligand molecules. This very standard and reproducible system should allow closer examination of the individual cytoadhesive events that occur between HPC and marrow stromal cells using cloned cell lines. PMID- 8425566 TI - Damage of tracer erythropoietin results in erroneous estimation of concentration in mouse submaxillary gland. AB - It has been previously reported that 1) plasma erythropoietin (Epo) titer during exposure to hypobaria is lower in nephrectomized rats and mice whose submaxillary glands (SMG) were either ablated or atrophied than in nephrectomized controls whose SMG were intact and 2) that the gland shows one of the highest levels of immunoreactive Epo (iEpo) in the body. The latter observation, however, was questioned recently when it was observed that SMG extracts degrade labeled Epo used as tracer antigen in the radioimmunoassay (RIA), thus giving invalid estimates of Epo. Since this interpretation was in turn questioned, the present study was conducted to obtain more information on the subject and make these conflicting points clear. Investigation of the reported/possible degradation of Epo by SMG homogenates was conducted via polyacrylamide gel electrophoresis followed by radioautography or by a RIA in solid phase in which there was no simultaneous incubation of the tracer antigen with the SMG homogenates. It was observed that 125I-labeled rhEpo was degraded when incubated with SMG homogenates. Degradation was rapid, being evident when incubation lasted 30 minutes, and occurred in the presence of a protease inhibitor. It showed a high degree of specificity since it did not occur when Epo was incubated with kidney homogenate or normal mouse serum. SMG homogenate did not degrade labeled thyrotrophic hormone and degraded alpha interferon (IFN-alpha) only partially. When estimates of iEpo in SMG homogenate were performed in conditions of simultaneous (SI-RIA) or nonsimultaneous (NSI-RIA) incubation of the homogenate with tracer Epo, it was observed that while estimates of Epo in plasma were similar in both types of RIA and somewhat higher in kidney homogenate in the SI RIA than in the NSI-RIA, estimates of Epo in SMG were about 60 times higher in the former than in the latter. Therefore, it could be concluded that most of the Epo detected by standard RIA in SMG homogenate does not represent true Epo because of damage of tracer Epo which determines loss of the integrity of the RIA system. PMID- 8425567 TI - In vitro purging of bone marrow with mafosfamide synergizes with in vivo chemotherapy to delay the hematological recovery in a murine model of autologous bone marrow transplantation. AB - Very long-lasting leukopenias and thrombocytopenias have been observed in patients submitted to transplantation of autologous bone marrow incubated in vitro with cyclophosphamide derivatives. With the aim of evaluating the contribution of in vitro exposure of bone marrow to mafosfamide (Asta-Z) and of in vivo chemotherapy given before bone marrow collection in these cytopenias, we designed a murine model of syngeneic bone marrow transplantation including treatment of donor and recipient mice with high doses of cyclophosphamide and in vitro exposure of the bone marrow transplant to Asta-Z. Blood platelets and leukocytes, medullary splenic colony forming unit (CFU-S), committed megacaryocytic (CFU-Meg) and granulomacrophagic (CFU-GM) precursor cell recovery was followed up to 56 days posttransplant. The data indicate that in vitro exposure of bone marrow to Asta-Z before reinfusion increases the delay in platelet recovery already induced by the chemotherapy given to donor mice and is specifically responsible for the prolongation of leukopenia. In recipient bone marrow, a synergy between the ablative effect of the in vitro treatment of bone marrow graft and the chemotherapy given to donors and recipients on CFU-S, CFU Meg and CFU-GM was found. PMID- 8425568 TI - Separation of red blood cells by field flow fractionation. AB - Field flow fractionation (FFF) is a new methodology described as being well suited for the separation and characterization of biopolymers and particles. On theoretical grounds, cells may be separated with FFF if they differ in size, density or deformability. In the present study, we first tried to determine optimal separation conditions for red blood cells; thereafter we used FFF to examine red cell changes during a phenylhydrazine-induced hemolytic anemia. It has been shown that in less than 30 minutes, FFF is able to separate normal red blood cells from Heinz body-rich cells or reticulocytes that differ in size or density. The successive steps of hemolysis and regeneration appear clearly on the fractograms. Advantages and drawbacks of the method are discussed. PMID- 8425569 TI - Regulation of the T cell receptor-alpha mRNA expression in the human lymphoblastic T cell line CEM. AB - We analyzed the transcriptional events involved in the T cell receptor (TcR) alpha mRNA expression in a human lymphoblastic T-cell line CEM. CD3-negative and CD3-positive CEM subclones that either lack mature TcR-alpha mRNA or express TcR alpha mRNA were used. Exposure of the TcR-alpha mRNA negative subclones to phorbol 12-myristate 13-acetate (PMA) was followed by 2- to 3-fold increase of transcription, indicating that PMA acts on a transcriptional level. No increase of transcription was observed after blocking protein synthesis with cycloheximide (CHX) or after sequential stimulation with CHX followed by PMA. On the posttranscriptional level, CHX as well as PMA induced a progressive stabilization of TcR-alpha mRNA in the nuclear compartment, which was independent of ongoing transcription. The half-life of the TcR-alpha mRNA upon stimulation was about 6 hours. The accumulation of mature TcR-alpha mRNA seemed to be controlled by nuclear events on a transcriptional as well as posttranscriptional level. The data imply that alterations of TcR-alpha gene transcription are dependent on protein synthesis. DNA-binding proteins enhance transcription and labile nuclear proteins target TcR-alpha mRNA for rapid turnover. PMID- 8425570 TI - Induction of a reversible block in murine CFU-C differentiation by exposure to nitrous oxide. AB - Patients subjected to prolonged exposure to nitrous oxide (N2O) often develop megaloblastic bone marrow changes. This toxicity is due to the N2O-mediated inactivation of cobalamin-dependent enzymes with resultant perturbations in cell metabolism. The effect of N2O on the behavior of murine colony-forming units cytokine (CFU-C) in vitro was studied by incubating granulocyte/macrophage colony stimulating factor (GM-CSF)-stimulated bone marrow cultures for 7 days in an atmosphere of either 5% CO2 in air or 50% N2O/5% CO2 in air. Exposure of bone marrow cells in agarose to N2O resulted in an approximately 50% reduction in colony formation when compared with cultures incubated in air. In contrast, when residual CFU-C numbers were determined in bone marrow liquid cultures after 7 days of incubation in the presence of GM-CSF, exposure to N2O was found to dramatically enhance CFU-C recovery. Since these liquid cultures contain a strong differentiation inducer, and are unable to support CFU-C generation, the enhancement of CFU-C recovery in N2O-exposed cultures appears to be related to its ability to induce a reversible block in CFU-C differentiation. The reversible block in CFU-C maturation seen in vitro parallels clinical observations where a rapid hematologic recovery is seen in N2O-exposed patients treated with hydroxycobalamin. These observations would suggest that N2O is not markedly cytotoxic to CFU-C and that its action is, at least in part, cytostatic in nature. PMID- 8425571 TI - Compensatory splenic hemopoiesis in beta-thalassemic mice. AB - beta-Thalassemic mice, homozygous for the deletion of the beta major-globin gene, were investigated for compensatory hemopoiesis in bone marrow and spleen. Apart from characteristic severe anemia, these mice have a marked granulocytosis, monocytosis and lymphocytosis. A large compensatory expansion of late (CFU-E) erythroid progenitor cells was demonstrated, predominantly in the spleen. Immature hemopoietic cells (CFU-S) were also expanded, as were early progenitor cells of erythroid (BFU-E), as well as granulocyte/macrophage (GM-CFU) and megakaryocytic (CFU-Meg) lineages. It is concluded that the persistent erythropoietic stress results in a selective expansion of immature hemopoietic cells and inappropriate production of nonerythroid blood cells from excess production of progenitor cells. PMID- 8425572 TI - Early divergence of erythroid lineage suggested by gene rearrangements in mouse hematopoietic neoplasms. AB - A total of 113 primary murine hematopoietic neoplasms, including those of erythroid, granulocytic, and T and B lymphoid lineages, were examined for rearrangement of immunoglobulin heavy (IgH) and kappa light chain (IgK) and T cell receptor beta and gamma (TcR-beta and TcR-gamma) genes. There was a total absence of Ig or TcR gene rearrangements in erythroid leukemias. In contrast, overlaps of IgH rearrangements were observed in myeloid and T cell as well as B cell neoplasms. In a minority of B cell lymphomas, rearrangements of TcR-beta or TcR-gamma genes were detected. This evidence of shared recombinase activity for myeloid, T cell, and B cell-lineage tumors and the absence of such activity in erythroid tumors suggest early divergence of the erythroid pathway. PMID- 8425573 TI - Interleukin-5 is the predominant eosinophilopoietin produced by cloned T lymphocytes in hypereosinophilic syndrome. AB - Cloned T lymphocytes (TLC) of the CD4+CD8- phenotype established from peripheral blood of a patient with idiopathic hypereosinophilic syndrome (HES) were found to release a lineage-specific eosinophilic colony-stimulating factor (Eo-CSF). The present study was undertaken to identify the lymphokine accounting for this Eo CSF activity. Comparison of TLC-derived Eo-CSF with recombinant human interleukin 5 (rhIL-5), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and recombinant human interleukin-3 (rhIL-3) by in vitro clonogenic assays revealed similar bioactivity of HES-derived Eo-CSF and IL-5. Neutralization studies using specific antibodies against IL-5, GM-CSF and IL-3 confirmed that IL-5 mainly accounts for the Eo-CSF activity in all 9 HES-derived TLC tested. Eosinophilic colony (CFU-Eo) formation supported by conditioned media of the TLC was significantly inhibited in all clones by addition of anti-IL-5 monoclonal antibody (MAB) to the conditioned media. Inhibition by anti-IL-5 MAB was specific and dose-dependent. In 2 of the 9 clones, anti-GM-CSF antibodies could partially neutralize the Eo-CSF activity in the conditioned media. In 4 clones, addition of a combination of anti-IL-5 MAB and anti-GM-CSF antiserum to the conditioned media reduced CFU-Eo formation significantly more than addition of anti-IL-5 MAB alone. In none of the TLC could a significant role for IL-3 in eosinophilic colony formation be shown. These results were confirmed at the mRNA level. Cytokine transcripts were detected by reverse transcription (RT) and subsequent polymerase chain reaction (PCR). Under the same experimental conditions, all HES-derived TLC, but only one third of tested TLC from healthy donors, expressed IL-5 mRNA 5 days after stimulation. In control TLC with inducible IL-5 mRNA expression, IL-5 transcripts were found for only 3 days after stimulation. In contrast, HES-derived TLC contained IL-5 mRNA at least until day 18 after restimulation. All HES clones expressed GM-CSF mRNA upon stimulation. Two HES-derived TLC were found to lack IL-3 mRNA even after stimulation. Whereas IL-5 was expressed abundantly in all HES-clones, the intensity of PCR products for GM-CSF and IL-3 showed striking differences. Our in vitro results suggest that IL-5 produced by activated CD4+ T lymphocytes plays a crucial role in the induction of eosinophilia in HES. In addition, GM-CSF but not IL-3 seems to contribute partially to the increased eosinophil production in HES. PMID- 8425574 TI - fgr proto-oncogene is expressed during terminal granulocytic differentiation of human promyelocytic HL60 cells. AB - In order to elucidate the function of the c-fgr protein tyrosine kinase, we have investigated the expression of c-fgr in the human promyelocytic cell line, HL60, during myeloid differentiation induced by dimethylsulfoxide (DMSO). The expression of c-fgr was preceded by growth arrest of DMSO-treated cells, as determined by [3H]-thymidine incorporation and colony-forming ability, and it became detectable when cells committed for terminal differentiation. The maximum expression was detected in the terminal stage of differentiation. The profile of tyrosine phosphorylation in cellular proteins was distinct among cells at various stages of the differentiation program. The 116 kd tyrosine-phosphorylated protein detected in exponentially proliferating HL60 cells diminished during the course of the growth arrest and a distinct profile of tyrosine phosphorylation (including 177 and 165 kd proteins) appeared in cells undergoing terminal granulocytic differentiation. These findings implicate the involvement of p55c fgr in the process of terminal granulocytic differentiation. However, the tyrosine kinase activity of p55c-fgr expressed in differentiating HL60 cells was markedly inhibited by the tyrosine phosphatase inhibitor, sodium orthovanadate, suggesting the presence of a mechanism involving tyrosine phosphorylation that negatively regulates the kinase activity of p55c-fgr. PMID- 8425575 TI - Megakaryocyte proplatelet-like process formation in vitro is inhibited by serum prothrombin, a process which is blocked by matrix-bound glycosaminoglycans. AB - The process of platelet shedding from megakaryocytes is incompletely understood, due in part to the impossibility of studying this dynamic process in vivo. Megakaryocytes in situ and in in vitro cultures display extended cytoplasmic processes constricted at platelet-sized intervals which presumably are the structural intermediates between megakaryocytes and platelets. This study describes the establishment of a serum-free culture system of purified guinea pig megakaryocytes in which extensive cytoplasmic process formation can be observed on 21 to 29% of the cells. The addition of as little as 0.05% pooled human serum to the cultures will completely but reversibly block process development. The serum inhibitor was identified as residual prothrombin, which upon contact with megakaryocytes is converted to the serine esterase thrombin. Thrombin directly prevents the formation of new processes and also induces retraction of existing processes. When megakaryocytes are cultured on Matrigel, process formation occurs even in an excess of thrombin. This potentiation of process development in the presence of inhibitory factors is mediated by the glycosaminoglycan content of Matrigel. The physiological implications of these observations are discussed. PMID- 8425576 TI - Rate of marrow engraftment does not appear to be adversely affected by early marrow infusion after total body irradiation. AB - Bone marrow transplantation (BMT) provides a means of increasing chemo radiotherapy doses beyond the limits imposed by marrow toxicity. The time interval between myeloablative therapy and marrow infusion needs to be optimized to ensure prompt engraftment. It has been suggested that early marrow infusion after completion of total body irradiation (TBI) may be detrimental to rapid marrow reconstitution. Hence a retrospective analysis of 75 BMT patients was performed to document the rate of marrow engraftment following marrow infusion given immediately after TBI. Engraftment rates (time to absolute neutrophil count to greater than 0.5 x 10(9)/L) of patients receiving different conditioning regimens were compared. Results show similar engraftment rates in patients receiving bone marrow infusion 27.7 and 32.1 hours after completion of chemotherapy in autologous (20.5 days) and allogeneic (18.5 days) transplants respectively, or 2.9 and 2.0 hours after completion of TBI in autologous (20.1 days) and allogeneic (19.4 days) transplants respectively. These results are similar to those reported for delayed marrow infusion after TBI and would support that early marrow infusion after TBI does not adversely influence rate of engraftment. Thus it appears appropriate to give marrow immediately after completion of TBI. PMID- 8425577 TI - Splenectomy does not enhance engraftment of DLA-nonidentical marrow transplants. AB - Previous studies in mice and monkeys suggested that either splenic irradiation of splenectomy led to enhanced allogeneic marrow engraftment. These findings suggested that a population of radiation-resistant host cells involved in mediating marrow graft rejection are sequestered in the spleen. In this current study, the effect of splenectomy in dogs receiving 9.2 Gy total body irradiation (TBI) and unrelated dog leukocyte antigen (DLA)-nonidentical donor marrow grafts was studied. We found that splenectomy did not significantly change the incidence of graft failure as compared to that observed in previously and concurrently transplanted nonsplenectomized recipients. PMID- 8425579 TI - The ERJ two years later. PMID- 8425578 TI - Thymomodulin increases HLA-DR expression by macrophages but not T-lymphocyte proliferation in autologous mixed leucocyte reaction. AB - Thymomodulin (TMD), a thymic biological response modifier, stimulates the release of tumour necrosis factor (TNF) and granulocyte-macrophage-colony stimulating factor (GM-CSF) in macrophage-lymphocyte cultures. We investigated the effects of the cytokines released in cultures with TMD, on the expression of human leucocyte antigen-DR (HLA-DR) antigens by alveolar macrophages (AM) and the T-cell proliferation induced in autologous mixed leucocyte reaction (AMLR) cultures or by T-cell mitogens. Among freshly isolated AM, 84 +/- 4% were HLA-DR positive, and this proportion was significantly reduced after 24 h cultures (60 +/- 3%, p < 0.05). In cultures without peripheral blood (PBL) lymphocytes, TMD did not change HLA-DR expression by AM CHLA-DR+AM); whilst in the presence of autologous PBL lymphocytes, TMD induced an increase in the proportions of HLA-DR+AM (TMD 100 micrograms.ml-1 79 + 3%, p < 0.04 vs control cultures). However, TMD did not change the ability of AM to induce T-cell proliferation in AMLR between AM and PBL lymphocytes. In contrast, in PBL mononuclear cell cultures, TMD induced a further increase of the cell proliferation due to the T-cell mitogens interleukin 2 (IL-2) or phytohaemagglutinin (PHA) (p < 0.05 vs each control culture with mitogens) or anti-CD3 antibodies (p < 0.03 vs control cultures). Thus, the cytokines released in cultures with TMD enhance macrophage HLA-DR expression. Whilst this phenomenon is not associated with changes in the ability of AM to stimulate T-cell proliferation, TMD is able to increase the mitogen-induced T cell proliferation. PMID- 8425580 TI - Changes in epithelial lining fluid albumin associated with smoking and interstitial lung disease. AB - Albumin is an important plasma protein which is useful in the assessment of in vivo membrane permeability in the lung. In subjects with interstitial lung disease (ILD) the relationship between albumin recovered from bronchoalveolar lavage (BAL) and other markers of inflammatory activity may provide useful information of the pathogenesis of the disease process. Unfortunately, its measurement is hampered by the variable dilution in BAL fluid. In this study, urea was used as a marker of epithelial lining fluid (ELF) dilution allowing the calculation of an apparent epithelial lining fluid volume and adjusted albumin content. We examined the relationship between ELF albumin content and BAL cell counts, immunoglobulin content, respiratory function tests and gallium lung scans in both smokers and nonsmokers with and without interstitial lung disease. Forty seven subjects with connective tissue disease and interstitial lung disease and 51 subjects with either connective tissue disease but no pulmonary involvement or non pulmonary malignancy (18 current smokers) underwent BAL, gallium lung scans and respiratory function tests. The subjects with ILD were further subdivided into those with active ILD or bronchiolitis using cluster analysis. In smokers without ILD an increased ELF volume and a decrease in ELF albumin were found. Increased ELF albumin was related to increased age. In subjects with ILD, increased albumin was strongly correlated with increased BAL lymphocyte absolute and differential counts, which overwhelmed any age or smoking effect. These findings suggest a possible causal relationship between lung vascular permeability and lymphocyte numbers in subjects with interstitial lung disease and reinforce the need to consider smoking and age as confounding factors in BAL analysis. PMID- 8425581 TI - Reversibility of exogenous corticosteroid-induced bone loss. AB - Osteoporosis is not usually considered to be reversible, as it is a consequence of the ageing process. However, an improvement of bone mineral density after successful surgery in Cushing's syndrome has been shown in several reports. The question of reversibility of exogenous corticosteroid-induced osteoporosis is, as yet, unanswered, possibly because of the difficulty in discontinuing steroids after long-term use. We describe six patients, all under 45 yrs of age, with chronic long-standing sarcoidosis, in whom long-term prednisone therapy resulted in 15 +/- 7% bone loss, as evaluated by quantitative computed tomography. This side-effect appeared fully reversible after prednisone withdrawal. This report of the reversibility of exogenous corticosteroid-induced bone loss needs confirmation in elderly people, where the capacity for recovery of bone mass could be reduced. Such potential for recovery may have implications for the pattern of use of corticosteroids. PMID- 8425582 TI - Regulation of antigen-presenting cell function(s) in lung and airway tissues. AB - A variety of cell populations present in respiratory tract tissues can express the function-associated molecules on their surface which are required for presentation of antigen to T-lymphocyte receptors. However, the potential role of individual cell populations in regulation of local T-lymphocyte-dependent immune reactions in the lung and airways depends on a variety of additional factors, including their precise localisation, migration characteristics, expression of T cell "co-stimulatory" signals, responsiveness to inflammatory (in particular cytokine) stimuli, host immune status, and the nature of the antigen challenge. Recent evidence (reviewed below) suggests that the induction of primary immunity (viz. "sensitisation") to inhaled antigens is normally controlled by specialised populations of Dendritic Cells, which perform a surveillance role within the epithelia of the upper and lower respiratory tract; in the pre-sensitised host, a variety of other cell populations (both bone-marrow derived and mesenchymal) may participate in re-stimulation of "memory" T-lymphocytes. PMID- 8425583 TI - Inhaled loop diuretics as potential new anti-asthmatic drugs. AB - The observation that changes in bronchial osmolarity can induce bronchoconstriction in asthma inspired the experimental studies which, unexpectedly, revealed that frusemide is an effective bronchoprotective agent against a variety of osmotic and non osmotic stimuli. Although the mechanism of this protective effect is not fully understood, studies in vivo and in vitro suggest that frusemide may inhibit the activation of different cell types induced by bronchoconstrictor stimuli. Other loop diuretics also exert bronchoprotective activity, but frusemide appears to be the more effective bronchoprotective agent of this family, regardless of their diuretic potency and lipid solubility. Despite the relatively large amount of experimental evidence, there is currently little information on the clinical effectiveness of frusemide in asthma and a long-term controlled study is currently in progress. The observations that treatment with a combination of inhaled acetylsalicylate and frusemide results in a markedly increased bronchoprotective effect compared to either drug alone, opens a new perspective in the possible clinical use of these drugs. Preliminary studies suggest that the association of these drugs is well tolerated and may result in a remarkable steroid sparing effect in patients with steroid dependent asthma, for whom a convenient alternative to long-term steroid therapy is not currently available. PMID- 8425584 TI - Lung adenocarcinoma with type II pneumocyte characteristics. AB - We report a case of primary lung adenocarcinoma with type II pneumocyte characteristics. Electron microscopic examination demonstrated that the tumour cells had well-developed microvilli and cytoplasmic lamellar inclusion bodies. These ultrastructural features are similar to those seen in type II pneumocytes of normal lung tissue. Western blot analysis, using monoclonal antibody against human surfactant protein A (SP-A), clearly demonstrated that the tumour cells expressed human SP-A, which is a major pulmonary surfactant protein produced by type II pneumocytes. These observations suggest that the tumour was derived from a type II pneumocyte. PMID- 8425585 TI - Primary malignant non-Hodgkin's lymphoma of the lung arising in mucosa-associated lymphoid tissue (MALT). AB - We describe a 10 yr follow-up of a patient with a primary malignant non-Hodgkin's lymphoma of the lung, arising in mucosa-associated lymphoid tissue (MALT). Although the patient was not treated with chemotherapy or radiotherapy, no peripheral spread occurred, confirming that MALT-associated lymphomas apparently remain localized until late in the course of the disease. PMID- 8425586 TI - Haemorrhagic pleural effusion several months after inhalation of a foreign body. AB - We present the case of a 50 yr old female with chronic obstructive lung disease, who, secondary to inhalation of a foreign body, developed recurrent pneumonia and a haemorrhagic pleural effusion (HPE). The foreign body (vegetable matter) was removed by rigid bronchoscopy and the patient recovered. PMID- 8425587 TI - Breathlessness in a teenager. PMID- 8425588 TI - Prevalence of asthma in patients with bronchiectasis in Hong Kong. PMID- 8425589 TI - Chlamydia pneumoniae infection in acute exacerbations of COPD. AB - Chlamydia pneumoniae, strain TWAR, is a frequent causative agent of acute respiratory disease. We assessed the incidence and prevalence of Chlamydia pneumoniae infections in COPD. We studied, from January 1990 to May 1991, 142 out patients with acute purulent exacerbations of chronic obstructive pulmonary disease (COPD) and 114 healthy control subjects. Oropharyngeal swab specimens were collected at each exacerbation and analysed using a high definition monoclonal indirect fluorescent antibody test for Chlamydia pneumoniae identification. Immunoglobulins G and M (IgG and IgM) fractions of antibodies to Chlamydia pneumoniae were studied by microimmunofluorescence test. Prevalence of specific IgG was 63% in COPD, and 46% in controls (Chi-squared test p = 0.007). Moreover, mean titre of IgG was significantly higher in COPD than in controls. Five patients were positive for specific IgM (> or = = 1:16), and one had a fourfold increase of IgG titre; four of these patients had been treated with ciprofloxacin 1 g.day-1 for 10 days, and two with erythromycin, 3 g.day-1 for 14 days, with remission of signs and symptoms of exacerbation. Chlamydia pneumoniae identification was always negative. Our data suggest that Chlamydia pneumoniae infection is a rather frequent event in COPD, since at least 4% of exacerbations may be associated with it. PMID- 8425590 TI - Restrictive treatment policy for pulmonary tuberculosis in a low prevalence country. AB - In Denmark, treatment of tuberculosis is generally recommended only if the diagnosis is confirmed bacteriologically. This policy may cause a delay in treatment, if the patients are smear negative. We investigated the duration of the treatment delay, and whether the delay would cause any serious health problems for the individual, or risk of contact infections, in a retrospective examination of 324 cases of pulmonary tuberculosis. The mean treatment delay was longer in the oldest age group. Concerning death due to delay, we found no risk for those patients who were not weakened by other disease or old age. Only 11 patients (3.6%) over the age 10 yrs were treated without bacteriological confirmation (1% for Danes). The infection risk from the smear negative but culture positive patients was minimal, as only one subject was definitely infected from a smear negative patient. However, a risk of transmission exists from patients who are initially culture negative but later become smear positive. In conclusion, we find the epidemiological and individual risks sufficiently low to continue our rather restrictive treatment policy. PMID- 8425591 TI - Reactive pleural inflammation caused by intratracheal instillation of killed microbes. AB - To investigate the pleural leucocyte response to severe alveolar inflammation, heat-killed Corynebacterium parvum were instilled intratracheally into the lungs of PVG rats and pleural lavage was performed. Polymorphonuclear neutrophils are not normally resident in the pleural space but were found transiently after intratracheal instillation of C. parvum. Macrophages increased gradually in the pleural space following instillation, reaching a peak at day 5. The activity of plasminogen activator inhibitor in the pleural leucocyte supernatants was increased at day 1, but returned to control levels by day 5. The activities of interleukin-1 and tumour necrosis factor secreted by pleural leucocytes were decreased compared with control pleural leucocytes at day 1 and were further reduced at day 5. The analysis of particle translocation showed that intratracheally instilled C. parvum and fluorescent beads with or without C. parvum did not reach the pleural space. We hypothesize that pleural inflammation resulting from C. parvum-induced inflammation in the lung is the result of transfer of a diffusible factor from the adjacent parenchyma. PMID- 8425592 TI - Patient selection and techniques for home mechanical ventilation. PMID- 8425593 TI - The effect of oral midazolam and diazepam on respiration in normal subjects. AB - Benzodiazepine have been shown to suppress ventilatory responses to hyperoxic hypercapnia (HCVR) and isocapnic HVR when taken parenterally. Most patients would, however, prefer to take an oral rather than parenteral preparation but the effect of oral benzodiazepine on these ventilatory responses has not been well studied. We therefore studied the effect of oral midazolam (7.5 mg) and diazepam (5 mg) both given orally on resting ventilation and respiratory drive, as assessed by HCVR and HVR. Flumazenil, a specific benzodiazepine antagonist, was administered intravenously to reverse the effect. A mental alertness-drowsiness index in five grades, from 1 (awake and alert) to 5 (asleep), was used to assess the sedation effect. Six normal male subjects, (aged 31 +/- 1.6 yrs) (mean +/- SD), participated in the study. Mean resting ventilation, and ventilatory response to HCVR and HVR were not significantly altered by these drugs when taken orally. Flumazenil also had not significant effect on HCVR and HVR. However the mental alertness-drowsiness index rose from 1 to 2.83 with oral midazolam and reversed to 1.25 with flumazenil. Similarly, this index increased from 1 to 2.25 after oral diazepam and reversed to 1.42 after flumazenil. In conclusion, we found that even though oral midazolam and diazepam produced a significant sedation effect, which was reversed with flumazenil, the drugs had no effect on ventilation at rest and the ventilatory responses to hypoxia and hypercapnia. PMID- 8425594 TI - Validation of automated sleep stage and apnoea analysis in suspected obstructive sleep apnoea. AB - Full-night polysomnography is necessary for the diagnosis of obstructive sleep apnoea (OSA). However, analysis of the sleep stages and apnoeas is time consuming. Computer systems for automated analysis have, thus, been developed to alleviate this task. We investigated 27 consecutive patients referred to our sleep laboratory with suspected OSA. The analysis of sleep stages and apnoeas was performed by visual scoring, according to Rechtschaffen and Kales, and by commercially available automated analysis device. The mean difference between visual scoring and automated analysis was -1, 111, -140, -3, 1 and 27 min, for sleep stages awake, I, II, III, IV and rapid eye movement (REM) respectively. For the apnoea index, the automated analysis rated a lower figure (mean difference 7.h-1, 95% confidence interval 2-12.h-1). The diagnosis of OSA was performed with a sensitivity of 85% and a specificity of 93% by automated analysis. Comparison of two independent handscores showed good agreement, with a mean difference of 6, 4, 3, -7, 1 and -1 min, for sleep stages awake, I, II, III, IV and REM, respectively. In conclusion, the automated analysis underestimates stage I sleep and the apnoea index. Visual scoring is advisable for control of the results. Automated analysis should only be used by those who are able to perform a visual analysis. PMID- 8425595 TI - Comparison of inspiratory effort in sniff-like aspiration reflex, gasping and normal breathing in cats. AB - The changes in airway occlusion pressure and airflow, occurring during two spasmodic breathing patterns, were studied and compared with normal breathing pattern in 12 anaesthetized cats. The inspiratory effort developed during the sniff-like aspiration reflex elicited by mechanical stimulation of the nasopharynx under control conditions proved to be very similar in character and intensity to the activity observed during gasping which occurred on resuscitation, of the same cats, from hypoxic apnoea. The starting (P50) and maximum (Pmax) airway occlusion pressure developed in these two spasmodic breathing patterns were very high. Extremely rapid rates of contraction and relaxation were detected by computer-assisted measurements of dynamic changes in both the pressure values and the slopes of pressure curves. The results suggest common effector mechanisms, reflecting similar forceful inspiratory drives, for the aspiration reflex and gasping. These two spasmodic processes differ substantially from normal breathing. Nevertheless, the aspiration reflex differs from gasping in that it can be elicited by activation of upper airway afferents during eupnea. Moreover, as yet, there is no definitive evidence that the brainstem mechanisms responsible for generating the aspiration reflex are the same as those of the gasp. The main benefits of this reflex are its rather easy elicitability under various conditions and its capability to induce important cardiorespiratory effects (e.g.) reversal of central apnoea) owing to its powerful activity. PMID- 8425596 TI - Effect of positive expiratory pressure on breathing pattern in healthy subjects. AB - The purpose of this study was to register breathing patterns, in healthy subjects, during breathing with a positive expiratory pressure. Integrated electromyographic (IEMG) activity of the following muscles was assessed: scalene muscle, parasternal muscle and abdominal muscles, using surface electrodes. Inspiration time, expiration time, total breathing cycle time, tidal volume and breathing frequency were measured using a water-sealed spirometer. Functional residual capacity was measured using a body plethysmograph. Oxygen uptake and carbon dioxide output were measured using an automatized ergometry set-up. All measurements were performed during undisturbed breathing and during breathing with positive expiratory pressures of 5 and 15 cmH2O. Phasic activity, but not tonic activity, of the scalene muscles and the abdominal muscles increased significantly during breathing with the expiratory pressures. No significant change was observed in phasic or tonic activity of the parasternal muscle. Mean (SD) tidal volume increased significantly from 0.8(0.2) l during undisturbed breathing to 1.1(0.3) l and 1.5(0.7) l during breathing with the expiratory pressures of 5 cmH2O and of 15 cmH2O, respectively. Respiration times, breathing frequency, oxygen uptake, carbon dioxide output and functional residual capacity remained unchanged. It can be concluded that, in healthy subjects, positive expiratory pressure increases tidal volume by activity of both expiratory and inspiratory muscles, while functional residual capacity remains unchanged. The changes appeared to be pressure dependent. PMID- 8425597 TI - Rheology of mucus and transepithelial potential difference: small airways versus trachea. AB - The transfer of water across the airway epithelium is closely related to the transepithelial potential difference (PD). Thus, PD should be directly involved in the regulation of airway intraluminal water content and, by extension, mucus rheology. Experiments by Boucher and co-workers (J Appl Physiol, 1980; 48: 169; and 1981; 51: 706) indicated that the values of PD in the small airways of dogs were considerably lower than in the trachea or mainstem bronchus. This fact suggests that water is increasingly removed from the airway lumen in the cephalad direction, and provides a possible mechanism whereby airway flooding is avoided as the total airway cross-section diminishes mouthward. We investigated this possibility by collecting and analysing mucus from the small airways and trachea of anaesthetized dogs and comparing our findings with measurements of PD. Mucus was collected on a cytology brush placed against the wall of the airway. Tracheal samples were taken from the lower lateral or anterior trachea, while small airway samples were taken from a 6th or 7th generation bronchus, chosen at random from either side. Measurements of PD were made at comparable sites. The mucus was analysed for its viscoelastic properties using the magnetic microrheometer technique. PD in the 6th-7th generation bronchus was significantly less than in the lower trachea (4.1 +/- 1.3 vs 17.2 +/- 7.1 mV). The rigidity of mucus collected from the small airways (log mechanical impedance (G*) at 100 rad.s-1) was significantly less than in the trachea (2.81 +/- 0.22 vs 3.01 +/- 0.29).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425598 TI - Relevance of Chlamydia pneumoniae in community-acquired respiratory infections. PMID- 8425599 TI - Flow cytometric DNA analysis of 20 bronchopulmonary neuroendocrine tumours. AB - Deoxyribonucleic acid (DNA) content of bronchopulmonary neuroendocrine tumours was measured by flow cytometry in order to investigate correlations between ploidy, S-phase fraction (SPF) and two histological classifications (World Health Organization (WHO), Warren and Gould), and clinical staging. A paraffin-embedded technique was used on 20 surgical specimens. Cases comprised (according to the classification of Warren and Gould) 7 carcinoids, 3 well-differentiated neuroendocrine carcinomas (WDNC), 6 intermediate neuroendocrine carcinomas (INC), and 4 small cell neuroendocrine carcinomas (SCNC). DNA aneuploidy was demonstrated in 3 out of 7 of the carcinoids, 3 out of 9 of the WDNCs and INCs, and 2 out of 4 of the SCNCs. A variable SPF was found in each group, except for the SCNCs which showed a constantly high SPF. In our small series, no correlation was noted between high SPF or aneuploidy and metastases. In conclusion, we observed no diagnostic value of malignancy for DNA aneuploidy. The SCNC group appeared to be an homogeneous group according to the SPF compared to the small cell carcinoma (SCC) group of the WHO classification. This and the prognostic incidence of high SPF need to be further studied. PMID- 8425600 TI - Upper airway reflexes and obstructive sleep apnoea. PMID- 8425601 TI - Histone deacetylase. A key enzyme for the binding of regulatory proteins to chromatin. AB - Core histones can be modified by reversible, posttranslational acetylation of specific lysine residues within the N-terminal protein domains. The dynamic equilibrium of acetylation is maintained by two enzyme activities, histone acetyltransferase and histone deacetylase. Recent data on histone deacetylases and on anionic motifs in chromatin- or DNA-binding regulatory proteins (e.g. transcription factors, nuclear proto-oncogenes) are summarized and united into a hypothesis which attributes a key function to histone deacetylation for the binding of regulatory proteins to chromatin by a transient, specific local increase of the positive charge in the N-terminal domains of nucleosomal core histones. According to our model, the rapid deacetylation of distinct lysines in especially H2A and H2B would facilitate the association of anionic protein domains of regulatory proteins to specific nucleosomes. Therefore histone deacetylation (histone deacetylases) may represent a unique regulatory mechanism in the early steps of gene activation, in contrast to the more structural role of histone acetylation (histone acetyltransferases) for nucleosomal transitions during the actual transcription process. PMID- 8425602 TI - Mechanism of pH-induced release of retinol from retinol-binding protein. AB - A hypothesis is proposed explaining the mechanism of pH-induced release of retinol from retinol-binding protein (RBP). A number of conservative positively charged side chains located on the retinol-binding face of the RBP molecule are involved in salt bridges with conservative negatively charged groups. At low pH these salt bridges are broken and the retinol-binding face of RBP holds from 8 to 12 positively charged groups, which can ensure a proper orientation of the RBP molecule relative to a negatively charged membrane, facilitating the release of retinol. The disruption of salt bridges and the electrostatic repulsion of positive charges can soften the structure of the molecule near the entrance to the retinol-binding pocket, which can trigger both the release of retinol and the transition of RBP to the molten globule state. PMID- 8425603 TI - Structure of the proteinase inhibitor eglin c with hydrolysed reactive centre at 2.0 A resolution. AB - The inhibition of serine proteinases by both synthetic and natural inhibitors has been widely studied. Eglin c is a small thermostable protein isolated from the leech, Hirudo medicinalis. Eglin c is a potent serine proteinase inhibitor. The three-dimensional structure of native eglin and of its complexes with a number of proteinases are known. We here describe the crystal structure of hydrolysed eglin not bound to a proteinase. The body of the eglin has a conformation remarkably similar to that in the known complexes with proteinases. However, the peptide chain has been cut at the 'scissile' bond between residues 45 and 46, presumed to result from the presence of subtilisin DY in the crystallisation sample. The residues usually making up the inhibiting loop of eglin take up a quite different conformation in the nicked inhibitor leading to stabilising contacts between neighbouring molecules in the crystal. The structure was solved by molecular replacement techniques and refined to a final R-factor of 14.5%. PMID- 8425604 TI - Variable and constant regions in the C-terminus of vinculin and metavinculin. Cloning and expression of fragments in E. coli. AB - Metavinculin differs from vinculin in having an additional insert of 68 to 79 amino acids in length in the C-terminal half of the molecule. Cross-species comparison of metavinculin sequences from pig, man, chicken and frog reveals a division of the insert into two parts: the first variable and the second highly conserved. The longest insert, 79 amino acids, was found in Xenopus laevis. Three different C-terminal constructs of vinculin and metavinculin over-expressed in E. coli could be purified by column chromatography. Two-dimensional gel electrophoresis and peptide analysis revealed pI values between 8.35 and 10.25 for the recombinant proteins. Biochemical and structural features of the metavinculin-specific sequence and the conserved vinculin/metavinculin carboxy terminus are discussed. PMID- 8425605 TI - Photoaffinity labelling of human leukotriene C4 synthase in THP-1 cell membranes. AB - Human leukotriene C4 synthase specific activity in the human monocytic leukemia cell line THP-1 (0.302 +/- 0.062 nmol LTC4 formed.min-1 x mg-1) was 7.6-fold higher than in U937 cells (0.040 +/- 0.017 nmol LTC4 formed.min-1 x mg-1) and comparable to dimethylsulfoxide-differentiated U937 cells (0.399 +/- 0.084 nmol LTC4 formed.min-1 x mg-1). Using the photoaffinity probe, azido[125I]-LTC4, a single polypeptide with a molecular mass of 18 kDa was specifically labelled in THP-1 microsomal membranes. The rank order of potencies for competition of azido[125I]-LTC4 photolabelling of the 18 kDa protein by glutathione, leukotrienes and their analogs was found to be LTC2 > (azido[127I]-LTC4 approximately LTC4) > (LTD4 approximately LTE4) > (LTA4 approximately LTB4) > S hexyl glutathione > glutathione, corresponded with the rank order of potencies for inhibition of LTC4 synthase activity but not inhibition of microsomal glutathione S-transferase activity. The 18 kDa protein specifically labelled by azido[125I]-LTC4 had high specificity for LTC4 and closely related leukotrienes and was separable from microsomal glutathione S-transferase. We conclude that azido[125I]-LTC4 specifically photolabels LTC4 synthase which is an 18 kDa polypeptide or contains an 18 kDa subunit. PMID- 8425606 TI - Effect of platelet activating factor on the kinetics of LDL oxidation in vitro. AB - Oxidatively modified low-density-lipoprotein (LDL) might contribute to the atherosclerotic process. This study was performed to examine an effect of platelet-activating factor (PAF) and of synthetic PAF analogs on Cu(II) induced oxidation of LDL in vitro: The D- and L-isomers of PAF and analogs with short chain sn-2-substituents, 1-O-alkyl-2-butyryl-sn-glycero-3-phosphocholine and 1-O alkyl-2-heptanoyl-sn-glycero-3-phosphocholine, were found to be the most effective inhibitors of LDL oxidation. Oxidation was inhibited completely at PAF concentrations above 100 microM. Lyso-PAF and analogs carrying longer chains at the sn-2 position were less effective. These results thus provide evidence for the involvement of other parameters in LDL oxidation beyond the content of natural antioxidants like vitamin E and beta-carotene. PMID- 8425607 TI - Complementation in situ of the yeast plasma membrane H(+)-ATPase gene pma1 by an H(+)-ATPase gene from a heterologous species. AB - In plants and fungi, the transport of solutes across the plasma membrane (pm) is driven by a proton pump (H(+)-ATPase) that produces an electric potential and a pH gradient. We expressed AHA2, a member of the Arabidopsis thaliana pm H(+) ATPase gene family, in yeast cells in which transcription of the endogenous pm H(+)-ATPase gene (pma1) had been turned off. AHA2 was expressed mainly in intracellular membranes and only supported very slow growth of transformed yeast cells. Removal of the last 92 C-terminal amino acids from the plant H(+)-ATPase produced an enzyme with 2-3-fold higher specific ATPase activity than the wild type plant enzyme. Surprisingly, the truncated H(+)-ATPase was now targetted to the yeast pm and fully supported normal yeast growth. PMID- 8425608 TI - Infrared studies of octopus rhodopsin. Existence of a long-lived intermediate and the states of the carboxylic group of Asp-81 in rhodopsin and its photoproducts. AB - The infrared absorption spectra of octopus rhodopsin and its photoproducts have been observed at 282K and 210K under irradiation of blue and orange light in a neutral condition. The acid metarhodopsin-minus-rhodopsin and lumirhodopsin-minus rhodopsin difference spectra have been obtained. A new intermediate (called transient acid metarhodopsin) with a lifetime of about 5 s has been found to exist prior to acid metarhodopsin. The present results, together with the data obtained previously, give information on the state of the carboxylic group in the side chain of Asp-81, which is the only acidic amino-acid residue in the part of opsin buried inside the membrane. This carboxylic group is protonated throughout the transformation series, but its state changes on going from transient acid metarhodopsin to acid metarhodopsin. It is probable that these two photoproducts are different from each other only in the opsin moiety. PMID- 8425609 TI - The region Ser333-Arg356 of the alpha-chain of human C4b-binding protein is involved in the binding of complement C4b. AB - Human C4b-binding protein (C4BP) functions as a cofactor to factor I in the degradation of C4b and accelerates the decay rate of the C4b2a complex. In this study we describe a monoclonal antibody directed against the alpha-chain of C4BP that inhibits the binding of C4b to C4BP. In order to identify the structural domain of the alpha-chain of C4BP that interacts with C4b, tryptic fragments of C4BP were generated. Amino acid sequence analysis of the fragments revealed that the residues Ser333-Arg356 of the alpha-chain of C4BP contain the epitope of this antibody, and as a consequence, that this part of the alpha-chain of C4BP is likely to be involved in the interaction with C4b. PMID- 8425610 TI - Synthesis of homocysteine thiolactone by methionyl-tRNA synthetase in cultured mammalian cells. AB - Homocysteine thiolactone is a product of an error-editing reaction, catalyzed by Escherichia coli and Saccharomyces cerevisiae methionyl-tRNA synthetases, which prevents incorporation of homocysteine into tRNA and protein both in vitro and in vivo. Here, homocysteine thiolactone is also shown to be synthesized by cultured mammalian cells such as human cervical carcinoma (HeLa), mouse renal adenocarcinoma (RAG), and Chinese hamster ovary (CHO) cells labeled with [35S]methionine, but not by normal human and mouse (Balb/c 3T3) fibroblasts. A temperature-sensitive methionyl-tRNA synthetase mutant of CHO cells, Met-1, does not make the thiolactone at the non-permissive temperature. The data indicate that methionyl-tRNA synthase is involved in synthesis of homocysteine thiolactone in CHO cells, thereby extending this important proofreading mechanism to mammalian cells. PMID- 8425611 TI - FVIIa derivatives obtained by autolytic and controlled cathepsin G mediated cleavage. AB - The heavy chain of coagulation factor VII contains a serine esterase entity. A partial cleavage in the heavy chain occurs during purification and activation of the single-chain zymogen, presumably as a result of autolysis. Neutrophil cathepsin G initially generates a Gla-domainless FVIIa without coagulant activity. However, on extended exposure cleavage also occurs in the heavy chain, resulting in a complete loss of enzyme activity. Four cleavage sites on the heavy chain, two susceptible to trypsin-like autolysis and two susceptible to chymotrypsin-like cathepsin G-mediated catalysis have been identified. The hydrolysis of peptide bonds in the heavy chain might contribute to regulation of the coagulation process in vivo. PMID- 8425612 TI - The rat insulin-degrading enzyme. Molecular cloning and characterization of tissue-specific transcripts. AB - The primary structure of the rat insulin-degrading enzyme (IDE) was determined by cDNA analysis. Rat IDE, as well as the previously characterized homologs from human and Drosophila, contain the carboxyl-terminal consensus sequence A/S-K-L, for peroxisome targeting. A stretch of 43 bp surrounding an alternatively used polyadenylation site is highly conserved between rat and human, suggesting that it may contain important regulatory information. Northern blot analysis revealed two IDE transcripts of 3.7 and 5.5 kb in various tissues. Testis was found to be exceptional in having three different RNAs (3.7, 4.1 and 6.1 kb) at a relatively high abundance. The expression of the IDE gene in testis is correlated with sexual maturation. PMID- 8425613 TI - Chemotactic and protease-inhibiting activities of antibiotic peptide precursors. AB - We have recently shown that two antimicrobial peptides (Bac5 and Bac7) and/or their immature forms (proBac5 and proBac7) can be released extracellularly from activated neutrophils. In the present study we have investigated the biological activities of the immature forms, which do not exhibit antimicrobial effects. We show that proBac7 is a monocyte-selective chemoattractant, potentially contributing to the recruitment of these cells to infection sites, whereas proBac5 efficiently inhibits the in vitro activity of cathepsin L, a cysteine proteinase thought to contribute to tissue injury in inflammation. PMID- 8425614 TI - Analysis of the catalytic center of cyclomaltodextrinase from Thermoanaerobacter ethanolicus 39E. AB - The amino acid sequences of cyclomaltodextrinase (CDase) from Thermoanaerobacter ethanolicus 39E (formerly Clostridium thermohydrosulfuricum 39E) and other amylolytic enzymes were compared by using linear alignment and hydrophobic cluster analysis. Two Asp and one Glu residue, which were considered to be the catalytic residues of the compared enzymes according to crystallographic or protein engineering experiments, were also conserved in CDase. Asp325, Asp421 and Glu354 of the CDase were individually replaced by means of site-directed mutagenesis. The mutant enzymes completely lost activity, suggesting that these residues play an important role in catalysis. PMID- 8425615 TI - Enhancing factor protein from mouse small intestines belongs to the phospholipase A2 family. AB - Enhancing factor (EF), a growth regulatory molecule, isolated from mouse small intestines, has been well characterized in this laboratory. It increases the binding of epidermal growth factor in a unique manner via its own receptor. In the first 20 N-terminal amino acids sequenced, EF showed 50% homology to human Group II phospholipase A2 (PLA2). Here we propose that EF is yet another, unidentified isoform of PLA2 which regulates cell proliferation via modulation of EGF binding. To our knowledge, this is the first report implicating PLA2-II-like molecules in growth regulation. PMID- 8425616 TI - ATP-driven Na+ transport and Na(+)-dependent ATP synthesis in Escherichia coli grown at low delta mu H+. AB - In inverted subcellular vesicles of Escherichia coli grown at high delta mu H+ (neutral pH, no protonophorous uncoupler), ATP-driven Na+ transport and oxidative phosphorylation are completely inhibited by the protonophore CCCP. If E. coli was grown at low delta mu H+, i.e. at high pH or in the presence of uncoupler, some oxidative phosphorylation was observed in the vesicles even in CCCP-containing medium, and Na+ transport was actually stimulated by CCCP. The CCCP-resistant transport and phosphorylation were absent from the unc mutant lacking F0F1 ATPase. Both processes proved to be sensitive to (i) the Na+/H+ antiporter monensin, (ii) the Na+ uniporter ETH 157, (iii) the F0 inhibitors DCCD and venturicidin, and (iv) the F1 inhibitor aurovertin. The CCCP-resistant oxidative phosphorylation was stimulated by Na+ and arrested by oppositely directed delta pNa. These data are consistent with the assumption that, under appropriate growth conditions, the F0F1-type ATPase of E. coli becomes competent in transporting Na+ ions. PMID- 8425617 TI - Current concepts in estrogen replacement therapy in the menopause. AB - OBJECTIVE: To provide a review of the risks and benefits of hormonal replacement therapy in the menopause, including new therapeutic regimens and modes of delivery. DESIGN: A review of the literature to identify published studies was accomplished using a computerized bibliographical search (Medline). RESULTS: Replacement therapy is effective in treating symptoms of estrogen deficiency and in lowering the risk of osteoporosis and cardiovascular disease. The daily administration of an estrogen and progestin eliminates the withdrawal bleed and increases patient compliance. This continuous form of therapy also consistently suppresses the endometrium, decreasing the risk of hyperplasia. More studies investigating the effect of continuous therapy on the lipid profile and cardiovascular disease are needed. CONCLUSIONS: New therapeutic regimens and modes of delivery decrease risk and increase patient acceptance of hormonal replacement therapy. PMID- 8425618 TI - Impact of the Human Fertilisation and Embryology Act (1990) on the practice of reproductive medicine in the United Kingdom. PMID- 8425619 TI - Implications of the Fertility Clinic Success Rate and Certification Act of 1992. PMID- 8425620 TI - Fertility drugs and ovarian cancer: red alert or red herring? PMID- 8425621 TI - Midluteal phase endometrial biopsy does not accurately predict luteal function. AB - OBJECTIVE: To investigate whether a midluteal phase endometrial biopsy accurately predicts luteal function. DESIGN: One nonpregnant menstrual cycle was evaluated in a prospective fashion. SETTING: Outpatient Clinic of the Clinical Center of the National Institutes of Health. PARTICIPANTS: Fifty healthy, normally cycling women. INTERVENTIONS: Serum progesterone (P) was measured daily throughout the luteal phase. An endometrial biopsy was performed 7 to 9 days after the luteinizing hormone (LH) surge, as detected by rapid plasma assays, and dated histologically according to Noyes' criteria. MAIN OUTCOME MEASURE: To correlate endometrial maturation with luteal P secretion. RESULTS: Mean integrated P measurements were reduced only when the lag between histologic and chronological dating was > or = 3 days or > or = 4 days, depending on whether chronological dates were assigned prospectively from the LH surge or retrospectively from the onset of next menses, respectively. However, these lags did not consistently predict deficient luteal function because subnormal integrated P secretion was seen in only 14% of women with these delays in endometrial maturation. CONCLUSIONS: Midluteal phase endometrial biopsy provides a crude test of luteal function that does not precisely distinguish luteal insufficiency. PMID- 8425622 TI - Efficacy and safety of repeated transcervical quinacrine pellet insertions for female sterilization. AB - OBJECTIVE: To investigate the rates of tubal occlusion, pregnancy, and side effects of repeated, monthly transcervical insertions of 252 mg quinacrine as pellets. DESIGN: Clinical trial among 159 reproductive age women receiving two monthly transcervical insertions of 252 mg of quinacrine followed by hysterosalpingograms (HSGs) 1 month after last insertion and an additional monthly insertion among women without evidence of bilateral tubal occlusion. Contraception of women's choice provided until bilateral tubal occlusion achieved, and surgical sterilization provided for women failing to achieve bilateral tubal occlusion after third quinacrine insertion. Women were followed for at least 24 months for evidence of pregnancy or side effects. RESULTS: Among the 159 women completing the protocol, 73% had evidence of bilateral tubal occlusion by HSGs after two insertions of quinacrine pellets and 94% after a third insertion. These 149 women were followed for 24 months without a pregnancy failure or serious side effect. CONCLUSION: Transcervical applications of quinacrine as pellets have potential for safe, effective, inexpensive, and easily deliverable female sterilization. PMID- 8425623 TI - Time-resolved fluoroimmunoassay compared with radioimmunoassay of luteinizing hormone. AB - OBJECTIVE: To determine if fluoroimmunoassay (FIA) of serum luteinizing hormone (LH) is more useful clinically than a conventional radioimmunoassay (RIA) because it has been suggested that FIA closely reflects biological activity. DESIGN: Comparison of serum LH measurements by RIA and FIA during various perturbations in normal ovulatory women and in women with polycystic ovarian syndrome (PCOS). SETTING: Normal ovulatory subjects were healthy volunteers and women with PCOS were untreated and newly diagnosed outpatients in our Reproductive Endocrinology/Infertility Clinic, Women's Hospital, at the Los Angeles County+University of Southern California Medical Center. PARTICIPANTS: Fifty three normal ovulatory women, ages 20 to 35, and 27 women with PCOS, ages 21 to 35. All were in good health and received no other medications during the study period. RESULTS: Fluoroimmunoassay of serum LH reflected status of known altered bioactivity better than with a conventional RIA. This was most evident during conditions of gonadotropin suppression and in patients with PCOS. An excellent correlation was found between values of FIA and RIA. CONCLUSIONS: The measurement of LH by FIA is clinically useful, specifically when a change in biological activity of LH is sought. PMID- 8425624 TI - Lack of impact of pharmacological growth hormone administration on circulating levels of reproductive hormones during the menstrual cycle in normal women. AB - OBJECTIVE: To evaluate the effects of pharmacological growth hormone (GH) administration on the secretion of pituitary and gonadal hormones during the menstrual cycle in normal women. DESIGN: Randomized, double-blind, placebo controlled crossover design. PATIENTS, PARTICIPANTS: Six normally menstruating, healthy women, 22 to 24 years of age and with a normal body mass index. INTERVENTIONS: Each participant was treated with placebo and growth hormone (12 IU/d) during two different 14-day periods, separated by a 6-week washout period. MAIN OUTCOME MEASURES: Serum insulin-like growth factor I (IGF-I), estradiol, progesterone, follicle-stimulating hormone, and luteinizing hormone. RESULTS: Administration of GH for 14 days resulted in a significant increase in serum IGF I, whereas no changes occurred in gonadotropin or sex steroid response. CONCLUSIONS: Growth hormone treatment during the first 14 days of menstrual cycle in normal women does not affect gonadotropin or sex steroid patterns. PMID- 8425625 TI - Tyrosine kinase activity of insulin-like growth factor I and insulin receptors in human endometrium during the menstrual cycle: cyclic variation of insulin receptor expression. AB - OBJECTIVE: To determine whether expression and binding or signaling characteristics of endometrial insulin-like growth factor I (IGF-I) and insulin receptors are modulated throughout the menstrual cycle. SETTING: Research laboratories of a university hospital and of the Institute for Diabetes Research. DESIGN: In vitro receptor binding and phosphorylation studies of human proliferative and secretory endometrial tissue and of cultured endometrial stromal cells. MAIN OUTCOME MEASURES: Binding and tyrosine kinase activation of IGF-I and insulin were studied in wheat germ agglutinin purified receptor protein. Binding data were analyzed by Scatchard plots. Autophosphorylation was measured by 32P incorporation into the 95 kd receptor beta-subunit; substrate phosphorylation was determined with poly(GluNa 4: Tyr 1). RESULTS: Binding studies revealed no differences of the affinities between cycle phases. Half maximal displacement of both receptors was approximately 1 nM in both phases. Insulin-like growth factor I receptor number appeared to be unaltered in both phases, whereas insulin receptor numbers and tyrosine kinase activity in the secretory phase were significantly increased. The increase of tyrosine kinase activity was entirely because of the increased receptor number as calculated from the binding data. In cultured endometrial stromal cells the increase of expression of insulin receptors could be induced by sexual steroids. CONCLUSIONS: On the receptor level IGF-I signaling to human endometrium is not modulated during the menstrual cycle, whereas insulin binding and signaling are likely to be enhanced in the luteal phase. The increased insulin receptor level may be required for normal luteal function. PMID- 8425626 TI - Regulation of androgen production in cultured human thecal cells by insulin-like growth factor I and insulin. AB - OBJECTIVES: To investigate if human thecal cells contain messenger ribonucleic acid (RNA) encoding insulin-like growth factor I (IGF-I) and insulin receptors and if IGF-I and insulin could stimulate androgen production in thecal cells. DESIGN: Poly-adenine+ RNA was extracted from fresh thecal tissue, and the expression of the genes encoding insulin and IGF-I receptors were analyzed. Isolated thecal cells were cultured 4 to 6 days with and without hormones. SETTING: Procedures were performed in a university laboratory. PATIENTS: Eight women in the follicular phase of natural cycles were undergoing gynecological laparotomy for reasons unrelated to ovarian pathology. The leading follicle(s) was excised, and dispersed cells of the theca interna layer were isolated through combined mechanical and enzymatic techniques. INTERVENTIONS: Luteinizing hormone (LH), IGF-I, and insulin were added to the cell cultures. MAIN OUTCOME MEASURE: The expression of IGF-I receptor and insulin receptor transcripts were analyzed by Northern blot. Medium levels of androstenedione and testosterone were measured by radioimmunoassay. RESULTS: In the separated thecal tissue both IGF-I receptor and insulin-receptor transcripts were detected. Insulin-like growth factor I and insulin potentiated LH-induced androgen secretion while having less pronounced effects on basal androgen production. CONCLUSION: The present study demonstrates that both insulin and IGF-I receptor genes are expressed and that insulin and IGF I can stimulate steroid production in human thecal cells. The study provides further support for the hypothesis that IGF-I and insulin may be involved both in physiological regulation of ovarian function as well as in its pathophysiology. PMID- 8425627 TI - Tumor necrosis factor-alpha alters steroidogenesis and stimulates proliferation of human ovarian granulosal cells in vitro. AB - OBJECTIVE: To determine if tumor necrosis factor alpha (TNF-alpha) altered human granulosa-luteal cell proliferation and steroidogenesis. DESIGN: Aspirates of follicles from women undergoing in vitro fertilization were subjected to Percoll gradients to collect an enriched population of granulosa-luteal cells. The granulosa-luteal cells were subjected to culture for a period of 10 or 20 days in the presence or absence of various doses of human recombinant TNF-alpha (0.1 to 10.0 ng/mL). PATIENTS: Granulosa-luteal cells from nine patients were evaluated for their response to TNF-alpha in vitro. Patients with three follicles > 16 mm and a serum estradiol (E2) concentration of > 1,836 pmol/L were selected for study. RESULTS: Tumor necrosis factor-alpha increased granulosa-luteal cell number. By day 10 of culture, 10 ng TNF-alpha/mL doubled cell number and > 95% of the cells exhibited 3 beta-hydroxysteroid dehydrogenase. Tumor necrosis factor alpha at 10 ng/mL increased progesterone (P) accumulation from day 4 through day 20 of culture. Tumor necrosis factor-alpha also increased E2 secretion but in a biphasic manner. During the first 14 days of culture, TNF-alpha increased E2, but thereafter E2 decreased to basal values by day 20. When steroidogenesis was expressed per 1,000 cells per days of culture, TNF-alpha did not increase P beyond controls but significantly increased E2 for the first 14 days of culture after which E2 per 1,000 cells declined. CONCLUSIONS: The results indicate that TNF-alpha stimulates granulosal-luteal cell growth and E2 secretion in vitro, and thus TNF-alpha may promote cellular events associated with formation of the corpus luteum; i.e., granulosa-cell proliferation and steroidogenesis. PMID- 8425628 TI - Comparison of intracervical, intrauterine, and intratubal techniques for donor insemination. AB - OBJECTIVE: To compare the efficacy of intracervical insemination (ICI), intrauterine insemination (IUI), and a combination of intratubal and intrauterine insemination (ITI/IUI) for donor insemination. DESIGN: Prospective randomized clinical trial. SETTING: The University of Michigan donor insemination program. PATIENTS, PARTICIPANTS: Forty-one women undergoing donor insemination with cryopreserved sperm for either isolated male factor or male factor plus ovulatory dysfunction corrected by clomiphene citrate. INTERVENTION: Each patient was randomly assigned to receive each of the three insemination techniques in consecutive cycles until pregnancy occurred or the patient dropped from the study. MAIN OUTCOME MEASURES: Cycle fecundity rates were compared using the chi 2 test, and cumulative pregnancy rates (PRs) determined by life table analysis were compared using a log-rank test. RESULTS: Cycle fecundity rate was significantly higher for IUI (18.3%) than for ICI (3.9%) or ITI/IUI (7.3%). By life table analysis, the cumulative PR for IUI was significantly higher than for ICI, but the PR for ITI/IUI was not. CONCLUSION: For donor insemination with cryopreserved sperm, IUI increases cycle fecundity compared with ICI. The addition of ITI to IUI, however, interferes with the apparent beneficial effect of IUI alone. PMID- 8425629 TI - Conception rate after in vitro fertilization in patients who conceived in a previous cycle. AB - OBJECTIVE: To evaluate whether a previously successful in vitro fertilization and embryo transfer (IVF-ET) cycle is a favorable prognostic factor for a subsequent cycle. DESIGN: A retrospective comparison between current IVF patients who have previously conceived in an IVF versus natural cycle. SETTING: The IVF unit of a university hospital. PATIENTS: Group A consisted of 51 patients (70 cycles of IVF ET) who previously conceived in an IVF-ET cycle, and group B included 141 patients (201 cycles of IVF-ET) who previously conceived in a natural cycle. All couples with male factor infertility were excluded. Ovulation induction protocol was identical for both groups and consisted of gonadotropin-releasing hormone agonist pretreatment followed by gonadotropin stimulation. MAIN OUTCOME MEASURES: Pregnancy rate per ET, cumulative pregnancy rate, and livebirth rate in both groups. RESULTS: The following parameters were comparable for both groups: age, menotropin dosage required for an adequate stimulation, ovarian response, mean number of oocytes retrieved per cycle, fertilization and cleavage rates, and the mean number of embryo transferred. Group A attained a significantly higher pregnancy rate (PR) than group B (31.4% versus 19.4%). Group A also achieved a significantly higher livebirth rate (22.9% versus 11.4%) than group B. Similarly, the cumulative PR curves and the cumulative livebirth rate curves for three consecutive IVF-ET cycles differed significantly between the two groups. CONCLUSION: A previous successful IVF cycle is a positive prognostic factor for a repeated IVF attempt. This effect could be because of either an improved endometrial response or a better embryo quality. It may be that this patient population is relatively immune to the known untoward effects of ovulation induction on endometrial development and, therefore, may represent a potential clinical model that can be used to further identify the factors influencing uterine receptivity after ovulation induction. PMID- 8425630 TI - Complete failure of fertilization in couples with unexplained infertility: implications for subsequent in vitro fertilization cycles. AB - OBJECTIVE: To determine whether complete failure of fertilization of oocytes in couples with unexplained infertility persists during subsequent in vitro fertilization (IVF) cycles. DESIGN: A retrospective study of 120 cycles of IVF in 44 couples with unexplained infertility and complete failure of fertilization during their first IVF trial. SETTING: In vitro fertilization unit of the Sheba Medical Center. PATIENTS: Forty-four couples undergoing IVF for unexplained infertility. MAIN OUTCOME MEASURE(S): Fertilization rate of retrieved oocytes and pregnancy rate. RESULTS: Of the initial 44 couples, 37 underwent additional IVF cycles and 30 (81.1%) achieved fertilization. Seven patients conceived during the study period as a result of IVF and two conceived spontaneously. CONCLUSIONS: Complete failure of fertilization does not necessarily persist during subsequent IVF cycles. Therefore, at least two more IVF cycles should be attempted before reverting to other therapeutic options. However, although fertilization can be achieved in most couples, the mean oocyte fertilization rate during subsequent cycles in this group is low. This suggests an underlying undiagnosed pathology of oocyte/sperm interaction in some of these patients. PMID- 8425631 TI - A controlled study to assess the use of in vitro fertilization with donor semen after failed therapeutic donor insemination. AB - OBJECTIVE: To assess the outcome of in vitro fertilization and embryo transfer (IVF-ET) with donor semen in patients who have failed to achieve conception with therapeutic donor insemination (TDI). DESIGN: Retrospective, controlled study. SETTING: All patients were managed and treated at the in vitro fertilization unit, John Radcliffe Hospital, Oxford, United Kingdom. PATIENTS, PARTICIPANTS: Thirty-eight patients undergoing 63 cycles of IVF with donor semen were compared with a control group of patients undergoing the same treatment during the same time period for tubal disease (the subgroup with the best success rates in our program), matched for age, type of infertility (primary or secondary), and number of attempts at treatment. MAIN OUTCOME MEASURES: We compared number of follicles produced, number of oocytes retrieved, fertilization rates, number of embryos obtained, number of embryos transferred, quality of embryos transferred, blastocyst formation in sibling embryos produced, clinical pregnancy rates (PRs), pregnancy loss, and pregnancy outcome. STATISTICS: Chi-squared test statistic with Yates' correction for continuity. RESULTS: There was a statistically significant difference between the number of livebirths in the donor IVF-ET group (37) and the control group (18). The cumulative PR after four cycles for the donor IVF-ET group (83%) was statistically greater than that of the control group (59%). There was no significant difference in all other parameters compared. CONCLUSION: Patients undergoing IVF-ET with donor semen have an excellent outcome. With the decline in PR after six cycles of TDI, early recourse to IVF-ET should be considered in this group of patients. PMID- 8425632 TI - Characterization of the first cell cycle in human zygotes: implications for cryopreservation. AB - OBJECTIVE: To study the temporal pattern of deoxyribonucleic acid (DNA) synthesis leading up to the first mitotic division in human one-cell stage zygotes. SETTING: In vitro fertilization program of a university hospital. PATIENTS: Couples donating spare embryos before the existence of an embryo freezing program. DESIGN: Incorporation of 3H-thymidine was examined between 9 to 27 hours after insemination in 72 untransferred human zygotes containing two pronuclei. Microscopic observations on an additional 978 transferred zygotes extended the 3H thymidine incorporation data. RESULTS: The first thymidine incorporation was seen 9 to 10 hours after insemination, and the heaviest incorporation occurred between 11 and 13 hours. Subsequently, thymidine labeling declined, and chromosomal condensation and cell division first occurred approximately 19 to 20 hours. At 20 hours after insemination, 89% of the zygotes had two visible pronuclei (PN). In contrast, by 24 hours, 41% had no visible PN, whereas 5% had cleaved to the two cell stage. By 27 hours, 38% had cleaved to the two-cell stage, and only 25% still had two visible PN. CONCLUSIONS: These results suggest that the DNA synthetic, S-phase of the human zygote is initiated by approximately 9 to 10 hours after insemination and is completed approximately 3 to 5 hours later. The duration of G2 phase and mitosis is in the range of 4 to 6 and 3 to 3.5 hours, respectively. Because zygotes may be particularly susceptible to damage during the S phase of the cell cycle, these findings suggest that the optimal time for freezing of human zygotes may be approximately 20 to 22 hours after insemination when the majority of zygotes should have entered the G2 phase, before pronuclear dissolution and chromosome condensation. PMID- 8425633 TI - Analysis of factors contributing to success in a program of micromanipulation assisted fertilization. AB - OBJECTIVE: To determine factors important to clinical success in micromanipulation-assisted in vitro fertilization (IVF). DESIGN: Procedures invoked in two separate series of micromanipulation-assisted IVF cycles, one unsuccessful (series I) and the other successful (series II), were compared in an effort to identify changes that led to clinical success. SETTING: University based IVF clinic. PATIENTS: In both IVF series involving micromanipulation, patients consisted of infertile couples who fit any of five categories of male factor related infertility. The female patients underwent controlled hyperstimulation for oocyte retrieval and the oocytes were inseminated normally or were subjected either to partial zona dissection or subzonal sperm insertion to assist fertilization. Results in all groups were compared between the two patient series. RESULTS: In the successful series II, a noticeable improvement in fertilization rate and embryo quality was observed compared with series I. A significant increase in the percentage of patients reaching embryo transfer, the pregnancy rate per transfer, and the pregnancy rate per retrieval were noted in series II; a 25% ongoing pregnancy rate per retrieval was observed overall in this successful group, with "ongoing" defined as manifestation of at least a fetal sac on ultrasound with no detectable problems. Patients with a mixed transfer of embryos derived from manipulated and normally inseminated oocytes had a 75% rate of pregnancy per transfer in series II. Differences between the two series could not be attributed to patient selection or biases in selection of oocytes relegated to micromanipulation. However, oocyte handling, micromanipulation, and culture protocols differed significantly between the two series in that temperature and pH of oocytes was better controlled, and micromanipulation time was minimized in series II. CONCLUSION: Success in micromanipulation depends on maintenance of the oocyte in a stable and supportive environment throughout the micromanipulation procedure. It is also important to minimize trauma to the eggs by performing micromanipulation rapidly and with minimal distortion of the egg. Patients with a poor fertilization rate in standard IVF may experience a substantial increase in the likelihood of pregnancy when micromanipulation-assisted fertilization is performed on some eggs. PMID- 8425634 TI - Maturational asynchrony between oocyte cumulus-coronal morphology and nuclear maturity in gonadotropin-releasing hormone agonist stimulations. AB - OBJECTIVE: To determine oocyte meiotic maturity and asynchrony between cumulus coronal morphology and nuclear maturity after gonadotropin-releasing hormone agonist (GnRH-a) and norethindrone-programmed stimulations. DESIGN: Oocyte meiotic maturity was evaluated at follicular aspiration in 4,961 oocytes after GnRH-a/follicle-stimulating hormone (FSH)/human menopausal gonadotropin stimulations (hMG) for in vitro fertilization patients and 299 oocytes after norethindrone-programmed clomiphene citrate (CC)/hMG in oocyte donors. Maturational asynchrony between the oocyte's cumulus-coronal morphology and nuclear maturity was evaluated in 2,336 oocytes. SETTING: In vitro fertilization program at the University of Iowa Hospitals and Clinics; academic tertiary care center. INTERVENTIONS: After evaluating oocyte cumulus-coronal maturity, cumulus masses were spread to determine oocyte nuclear maturity. RESULTS: Fourteen percent, 17%, 50%, 17%, and 2% of oocytes were prophase I, metaphase I, metaphase II, postmature metaphase II, and atretic, respectively. Asynchrony was noted in 28% of prophase I, 71% of metaphase I, 11% of metaphase II, 45% of postmature metaphase II, 32% of atretic, and 28% of all oocytes. Significant differences were not found between GnRH-a and norethindrone-programmed stimulations in asynchrony between cumulus-coronal morphology and nuclear maturity or percentage of prophase I, metaphase I, metaphase II, postmature metaphase II, or atretic oocytes. Sixty-seven percent of oocytes possessed a polar body at retrieval. The rate of fertilization was significantly higher for metaphase II oocytes than postmature metaphase II and metaphase I oocytes > prophase I oocytes. Parthenogenetic activation tended to be highest for postmature metaphase II oocytes. Embryo cleavage was significantly higher for postmature metaphase II, metaphase II, and metaphase I oocytes than for prophase I oocytes. CONCLUSIONS: This is the first report of asynchrony between cumulus-coronal morphology and nuclear maturity at follicular aspiration in GnRH-a and norethindrone-programmed stimulations. Asynchrony was observed in 28% of oocytes. A higher percentage of oocytes possessed a polar body at egg retrieval with these stimulation regimens compared with rates reported previously for FSH, FSH/hMG, and CC/hMG stimulations. PMID- 8425635 TI - Sex preselection through albumin separation of sperm. AB - OBJECTIVE: To determine if passage of sperm through columns of liquid albumin before their use in artificial insemination could affect the sex ratio at birth. DESIGN: Sperm were isolated by layering over columns of liquid albumin. The isolated fractions were inseminated into the uterus on the presumptive day of ovulation. SETTING: Patients were treated in 65 clinical practices (Sperm Centers) located both in this country (57) and abroad (8). PATIENTS: Individuals were self-selected by their desire to have a child of a specified sex. MAIN OUTCOME MEASURE: The sex of offspring resulting from the insemination of isolated sperm. RESULTS: Insemination with sperm isolated to enhance male sex preselection produced 71% and 76% males depending on the technique. Use of isolated sperm in women who were taking clomiphene citrate was associated with a 69% birth of females. CONCLUSIONS: Separation of sperm on columns of liquid albumin can affect the sex ratio at birth. PMID- 8425636 TI - The sex ratio of normal and manipulated human sperm quantitated by the polymerase chain reaction. AB - OBJECTIVE: To establish the primary sex ratio, the relative abundance of X and Y chromosome-bearing sperm, in unselected sperm and in sperm selected by swim-up or Sephadex filtration (SpermPrep column; Fertility Technologies, Inc., Natick, MA). This was done to evaluate the possibility that these semen manipulations change the primary sex ratio. DESIGN: Ninety-eight unmanipulated semen samples were analyzed for sex chromosome content using quantitative polymerase chain reaction (PCR). A smaller number of samples was analyzed before and after either swim-up or Sephadex filtration. RESULTS: The mean percentage of all sex chromosomes identified as a Y chromosome in unmanipulated semen samples ranged from 41.9% to 56.7%, with a mean average of 50.3%. There was no significant change in sex chromosome composition after either swim-up (n = 17) or column filtration (n = 20). CONCLUSIONS: The chromosome compositions of semen samples from a large number of men have equal numbers of X and Y. Swim-up and SpermPrep filtration do not appear to alter the primary sex ratio. PMID- 8425637 TI - Evaluation and treatment of ejaculatory duct obstruction in the infertile male. AB - OBJECTIVE: To evaluate the role of ejaculatory duct obstruction as a cause for male factor infertility and review the treatment of this condition. DESIGN: A retrospective study was carried out involving male patients diagnosed as having ejaculatory duct obstruction as a contributing cause for their subfertility. SETTING: Patients were evaluated and treated through a university-based male infertility clinic. PATIENTS, PARTICIPANTS: Patients were evaluated for the presence of ejaculatory duct obstruction if they suffered from decreased ejaculate volume, sperm density, and sperm motility but had normal physical examinations and otherwise normal laboratory evaluations. INTERVENTIONS: Patients were treated by transurethral resection of the ejaculatory ducts. MAIN OUTCOME MEASURES: Results of semen analysis and pregnancy rates. RESULTS: An alteration in semen quality was achieved in 79% of patients after transurethral resection. An increase in sperm density or motility was achieved in 50%, whereas 29% showed an increase in ejaculate volume only. Pregnancy rate postoperatively was 29%. CONCLUSIONS: Ejaculatory duct obstruction as a cause of male infertility is more common than was previously recognized, especially among nonazoospermic patients. In properly selected patients, transurethral resection of the ejaculatory ducts can result in marked improvement in semen quality with subsequent pregnancy. PMID- 8425638 TI - Subclinical alterations in hormone and semen profile in athletes. AB - STUDY OBJECTIVE: To investigate the effects of two forms of exercise, endurance training (running) and resistance training (weight lifting), on reproductive function in male athletes. DESIGN: Cross-sectional study. SETTING: Reproductive Endocrinology and Exercise Laboratory. SUBJECTS: Twenty-eight healthy male volunteers, 18 to 35 years of age, including 10 endurance-trained runners, 8 resistance-trained weight-lifters, and 10 sedentary controls. MAIN OUTCOME MEASURE(S): Hormonal evaluation included determination of plasma levels of total testosterone (T), serum levels of free T, luteinizing hormone (LH), follicle stimulating hormone, prolactin, and estradiol, and urinary excretion of LH. Semen analyses included an evaluation of sperm characteristics in terms of density, count, motility, and morphology, and a determination of in vitro sperm penetration of standard bovine cervical mucus. RESULTS: Compared with sedentary controls, endurance-trained and resistance-trained athletes presented with significantly lower levels of total and free T. There were no significant differences in the serum levels of all other circulating and urinary hormone measurements among the three groups. Sperm density, motility, and morphology were significantly altered only in the endurance-trained runners. In vitro sperm penetration of standard cervical mucus was significantly reduced in the endurance trained runners. CONCLUSION: Both endurance and resistance training modify the male reproductive hormone profile in a similar manner; however, only endurance training, in the form of running, is associated with subclinical modifications in semen characteristics. PMID- 8425639 TI - The contribution of a hidden male factor to unexplained infertility. AB - OBJECTIVE: To assess sperm function in patients with unexplained infertility in comparison with normal fertile men. DESIGN: Prospective study. SETTING: Infertility outpatient clinic and donor insemination program, University Department of Obstetrics and Gynecology at the Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: Nineteen patients with unexplained infertility and nineteen normal fertile men from a donor insemination program. MAIN OUTCOME MEASURES: Monitoring of reactive oxygen species generation; computerized assessment of the hyperactivated motility pattern of the spermatozoa under capacitating conditions, with and without the addition of follicular fluid (FF); evaluation of sperm-zona pellucida (ZP) binding. RESULTS: Reactive oxygen species generation was not significantly different between the study and control group. Follicular fluid-induced hyperactivation (HA) was significantly lower in patients with unexplained infertility than in the donors, but no difference was found comparing spontaneous HA (without FF) between the two groups. Follicular fluid-induced HA was completely absent in 39% of infertile men and only one fertile man. A low sperm-ZP binding ratio (< or = 0.3) was observed in 28% (5/18) of patients with unexplained infertility. In all, nine infertile men (47%) had 0% FF-induced HA and/or low sperm-ZP binding ratio. CONCLUSIONS: Assessing FF induced HA and sperm-ZP binding may have clinical significance in distinguishing a subgroup of men with unexplained infertility with sperm function abnormalities in which the fertilizing capacity of the spermatozoa might be reduced. These patients cannot be identified by the conventional semen analysis. Both tests may also be useful in planning the appropriate treatment for couples with unexplained infertility. PMID- 8425640 TI - Effects of pentoxifylline on human sperm motility in normospermic individuals using computer-assisted analysis. AB - OBJECTIVE: To determine if pentoxifylline has an in vitro effect on human sperm motility in subjects with normal semen analysis. DESIGN: Prospective, longitudinal study of the effect of pentoxifylline on normospermic semen samples using computerized analysis. SETTING: The Assisted Conception Unit, Royal Maternity Hospital, Belfast. SUBJECTS: Eight healthy male volunteers of proven fertility within 2 years of study and whose general health was unchanged in the intervening period. RESULTS: Pentoxifylline did not consistently increase the number of progressively motile sperm or their straight line velocity (VSL). In marked contrast, the curvilinear velocity (VCL) was increased significantly at all times, up to and including 240 minutes of exposure to the drug. This resulted in a substantial reduction in the linearity (VSL/VCL) of sperm progression. The most dramatic effect that pentoxifylline had on sperm movement was to increase the amplitude of lateral head displacement (ALH) over the same time interval. This change was accompanied by an increase in the beat cross frequency. CONCLUSIONS: The addition of pentoxifylline to prepared sperm samples from normospermic men does not increase the number of progressively motile sperm nor the progressive velocity. However, it does significantly alter the quality of certain characteristics of already motile sperm. PMID- 8425641 TI - Earlier testing after vasectomy, based on the absence of motile sperm. AB - OBJECTIVE: To review the results of postvasectomy testing when clearance was based on the absence of motile sperm and to compare them with regimens based on complete azoospermia. DESIGN: A review of 2,260 seminal assay results from 3,178 consecutive vasectomies performed during a 17-year period. SETTING: An outpatient vasectomy service in a private group practice in suburban Sydney. RESULTS: Clearance was given sooner and with less testing than with other reported regimens, without loss of reliability. CONCLUSIONS: Testing can be done 4 weeks after vasectomy, regardless of the number of postvasectomy ejaculations. If specimens are examined within 12 hours of collection, clearance may safely be given if motile sperm are absent. Repeat tests are essential if any motile sperm remain but are not needed if only nonmotile sperm are found. PMID- 8425642 TI - Time-related effects of RU486 treatment in experimentally induced endometriosis in the rat. AB - OBJECTIVE: To evaluate the time-dependent effect of the progesterone antagonist RU486 on experimentally induced endometriosis in the rat. DESIGN: Endometriosis was induced by implanting endometrium onto the peritoneal side wall. Size of the implants was measured before and after treatment. PARTICIPANTS: Fifty-two adult female Sprague-Dawley female rats weighing 200 to 225 g. SETTING: Departmental research laboratory of university hospital. INTERVENTION: Animals treated with RU486 or vehicle only for 2, 4, 6, or 8 weeks. MAIN OUTCOME MEASURES: Size of the implant was measured before and after treatment; relative change in size of the implants was calculated. RESULTS: There was not a statistically significant difference between the treatment groups and the control groups. There was no regression of endometriosis in any of the treatment groups. CONCLUSIONS: RU486 administered for 8 weeks in the rat model was not effective therapy for endometriosis. PMID- 8425643 TI - Long-term management of adenomyosis with a gonadotropin-releasing hormone agonist: a case report. AB - Leuprolide acetate was used to produce a constant hypoestrogenic environment in a young patient with histologically confirmed adenomyosis. Conservative medical therapy was initiated because of the patient's complaint of severe dysmenorrhea coupled with her strong desire for uterine conservation. The initial daily subcutaneous dose was eventually converted to monthly intramuscular depot formulation for patient convenience. A dramatic therapeutic response was observed with each course of therapy. This included a marked reduction in uterine size, amenorrhea, and complete resolution of pelvic pain. Cyclic use of an OC agent following LA was associated with a return of symptoms and uterine growth. The patient did, in fact, conceive immediately on cessation of analogue therapy. PMID- 8425644 TI - Pregnancy after medical therapy of adenomyosis with a gonadotropin-releasing hormone agonist. AB - A patient with long-standing secondary infertility was explored for myomectomy, at which time severe adenomyosis was found. A 6-month course of nafarelin acetate resulted in resolution of dysmenorrhea and uterine enlargement. The patient conceived quickly. Although the patient spontaneously aborted, this report presents the first in which medical therapy of adenomyosis is associated with successful treatment of infertility. PMID- 8425645 TI - Pregnancy during long-term gonadotropin-releasing hormone agonist therapy associated with clinical pseudomenopause. AB - A case of pregnancy occurring with escape of suppression from GnRH-a therapy is described. Clinical pseudomenopause had been established, and the pregnancy was exposed to depot GnRH-a for 8 weeks. Normal pregnancy outcome demonstrated that successful implantation may occur in the first escaped ovulatory cycle. PMID- 8425646 TI - Adverse effects of leuprolide acetate depot treatment. AB - One hundred two premenopausal women with uterine myomas were treated with LA depot. All women experienced adverse effects during GnRH-a treatment. The majority experienced symptoms attributed to a rapid and sustained decrease in circulating estrogen concentrations beginning 2 to 7 weeks after initiation of treatment. A smaller percentage of women experienced transient adverse effects due to the initial surge and subsequent rapid decline of gonadal steroids. The mechanism of other adverse effects (e.g., arthralgias) remains unclear. Only 6% of women terminated LA treatment before 12 weeks because of the severity of adverse effects. PMID- 8425647 TI - The effect of maternal serum on phase-change temperature in human embryo cryopreservation. AB - The effect of addition of maternal serum to cryomedium on the phase-change temperature for embryo cryopreservation has been investigated. No effect on the eutectic point (i.e., earliest seeding temperature) was found within patients because of serum concentration or the time in the ovarian cycle when the serum was collected. However, the optimum seeding temperature did vary between patients, which suggests that self-seeding devices for embryo cryopreservation may be practically beneficial. PMID- 8425648 TI - The effects of baseline ovarian cysts on cycle fecundity in controlled ovarian hyperstimulation. AB - Patients undergoing COH were prospectively studied in 174 cycles for the presence of baseline ovarian cysts. In 37.4% of all cycles, a baseline cyst > 10 mm mean diameter was found, but a cyst was more common in subsequent cycles than on the first (41.5% versus 15.8%). Cycle fecundity as determined by life table analysis was significantly higher if no baseline cyst were present (0.25 versus 0.06, P > 0.01). These findings suggest that baseline ovarian cysts may adversely affect the chances for pregnancy in COH not associated with IVF or GIFT. PMID- 8425649 TI - Histocompatibility in in vitro fertilization couples. AB - Major histocompatibility differences between mother and fetus may facilitate implantation and maintenance of pregnancy. Thus, we have investigated the compatibility of HLAs in couples with three successive failed IVF-ET cycles. The study couples (n = 15) shared a statistically greater number of HLAs than IVF couples achieving a viable pregnancy with their first IVF-ET attempt (n = 15) and a control group of 100 fertile couples. No difference between fertile and infertile control couples was observed regarding HLA sharing. Thus, we conclude that some cases of unsuccessful ETs after IVF might be caused by underlying close histocompatibility between partners. PMID- 8425650 TI - Ovarian torsion in a premenopausal woman treated with tamoxifen for breast cancer. AB - Tamoxifen therapy in premenopausal women with breast cancer may be associated with the development of ovarian cysts. In this report, a case of ovarian torsion associated with TAM therapy is described. PMID- 8425651 TI - Effects of pentoxifylline on sperm motility and hyperactivated motility in vitro: a preliminary report. AB - Previous reports (2, 3) have suggested that pentoxifylline increases sperm motility. In this preliminary report based on five asthenozoospermic and five normal motility semen samples, we were unable to demonstrate any statistically significant effect of pentoxifylline on percent motility of human spermatozoa. However, in vitro exposure to capacitation medium with pentoxifylline may lead to an increase in total hyperactivated motility in asthenozoospermic samples, an effect not evident in the normal motility samples in this study. PMID- 8425652 TI - Ovarian transposition with subsequent intrauterine pregnancy. AB - A 28-year-old married, Korean woman was diagnosed with a right unicornuate uterus, a left hypoplastic tube, and a right ovarian necrotic corpus luteal cyst. We performed a right oophorectomy and an excision of the left hypoplastic tube and then transferred the left ovary to the right broad ligament with intact vascular pedicles. Thus the remaining left ovary and right fallopian tube were placed in proximity. The patient conceived 2 months after the operation and delivered a living male infant. PMID- 8425653 TI - Computer-assisted assessment of hamster ovarian follicle pools: a novel approach. AB - With computerized image capture, primordial and primary follicles were easily and reproducibly measured. This technology appears ideal for following rarely studied small follicle populations and may have utility in the direct evaluation of superovulation protocols and other drug therapy. PMID- 8425654 TI - Experimental use of CUSA--value? PMID- 8425655 TI - Dynamics of fertility stress--unique? PMID- 8425656 TI - Extent and depth of adenomyosis--assessable? PMID- 8425657 TI - Guidelines for gamete donation: 1993. The American Fertility Society. PMID- 8425658 TI - Can we really predict IDDM? AB - Risk of progression to IDDM has been assessed extensively in first-degree relatives of IDDM patients, and highly specific prediction is possible within a small subset of this population. Because approximately 90% of future cases will come from those who have no close relative with IDDM, prediction and intervention within the general population will become the main priority for the future. This review presents a decision tree analysis of risk of progression to IDDM, highlights the different prognosis of markers when applied to those with and without a family history of the disease, and proposes a strategy for disease prediction in the latter. Large collaborative studies in well-characterized populations will allow new predictive markers and models to be evaluated, and strategies of intervention to be tested with maximum efficiency and minimal delay. PMID- 8425659 TI - Elevation of factor XIa-alpha 1-antitrypsin complex levels in NIDDM patients with diabetic nephropathy. AB - Excess activated FXIa in plasma indicates hypercoagulability in the early contact phase. We have already developed methods for detecting the hypercoagulable state in clinical samples by our ELISA for complexed FXIa and alpha 1AT, which has been confirmed to be the predominant inhibitor of FXIa. In diabetes, whether the activation of FXI is associated with the development of vascular complications remains unknown, although various hemostatic abnormalities have been described. We tested the complexed FXIa-alpha 1AT level in 45 NIDDM patients, who were divided into three groups according to the development of diabetic nephropathy, as assessed by UAE. Normoalbuminuria was defined as UAE < 15 micrograms/min, microalbuminuria as UAE in the range of 15-200 micrograms/min, and albuminuria as UAE > 200 micrograms/min. In the patients as a whole, FXIa-alpha 1AT and TAT levels were significantly increased compared with these levels in age-matched control subjects (17.3 +/- 5.7 vs. 12.4 +/- 2.4 ng/ml and 2.67 +/- 1.23 vs. 1.93 +/- 0.45 ng/ml, respectively). No significant difference was observed between FXIa-alpha 1AT levels in the control subjects and in the normoalbuminuric group (13.0 +/- 2.1 ng/ml; n = 19). However, in the microalbuminuric (17.9 +/- 3.9 ng/ml; n = 16) and albuminuric (24.1 +/- 5.4 ng/ml; n = 10) groups, FXIa-alpha 1AT levels were significantly increased compared with those in the control and normoalbuminuric group. The TAT level was not correlated with FXIa-alpha 1AT, and no significant differences in its levels were found among these diabetic groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425660 TI - Driving decrements in type I diabetes during moderate hypoglycemia. AB - Diabetic hypoglycemia produces cognitive-motor slowing, which is assumed to increase risk of automobile crashes. This study investigated driving decrements during and after hypoglycemia, and the patients' awareness of driving decrements. We used a randomized, single-blind, crossover design and conducted the study at the University of Virginia's General Clinical Research Center. We studied a volunteer sample of 27 consecutive adult type I diabetic patients who responded to newspaper ads. Two dropped out (final n = 25). Mean age (+/- SD) was 35.9 +/- 14 yr. Diabetes history was 14.6 +/- 10.5 yr, with HbA1 of 10.8 +/- 2.9%. Driving experience was 19 +/- 13.2 yr. Participants drove a state-of-the-art driving simulator on two consecutive days: Control day involved four euglycemia (mean blood glucose, 6.3 +/- 0.89 mM) driving tests; experimental day involved testing at euglycemia, mild hypoglycemia (mean blood glucose, 3.6 +/- 0.33 mM), moderate hypoglycemia (mean blood glucose, 2.6 +/- 0.28 mM), and again at euglycemia. Patients were blind to blood glucose manipulations and levels. Driving performance was not disrupted at mild hypoglycemia nor after recovery from moderate hypoglycemia. Moderate hypoglycemia disrupted steering, causing more swerving (P < 0.03), spinning (P < 0.03), time over midline (P < 0.05), and time off road (P < 0.01). It also resulted in an apparent compensatory slowing, with more very slow driving (P < 0.04). Global driving performance decrements were observed in 35% of the patients, only 50% of whom stated they would not drive under similar conditions. Driving decrements were unrelated to demographic, disease, or driving history variables.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425661 TI - Reduced sample number for calculation of insulin sensitivity and glucose effectiveness from the minimal model. Suitability for use in population studies. AB - The FSIGT has been extensively applied to the minimal model of glucose kinetics to obtain noninvasive measures of Sl. The protocol has been modified by the addition of a bolus tolbutamide or insulin injection 20 min after glucose. Although the modified protocol has improved the Sl estimate, the method still requires a relatively large number of samples (n = 30). To reduce the total number of samples, we choose a sample schedule that minimizes the variance of the parameter estimates and the error in reconstructing the plasma insulin profile. With data from 10 subjects (BMI 30 +/- 7 kg/m2; Sl 0.9-10.2 x 10(-4) min-1.microU 1 x ml-1), a schedule consisting of 12 samples (0, 2, 4, 8, 19, 22, 30, 40, 50, 70, 90, and 180 min) was obtained. Estimates of Sl obtained from the reduced sampling schedule were then compared with those obtained with the full sampling schedule. In all 10 individuals, the Sl estimates were almost identical. A second, much larger data base consisting of 118 modified FSIGTs performed in 87 subjects (67 men, 20 women; BMI from 19.6 to 40 kg/m2 for men and 26.7 to 52.5 for women; Sl from 0.35 to 14.1 x 10(-4) min-1 x microU-1 x ml-1) was then used to independently assess the efficacy of the reduced sampling protocol. For this data base, the correlation between Sl, which was calculated from the full versus the reduced sampling schedule, was 0.95. The mean relative deviation was -1.5% (not significantly different from zero), and the SD of the relative deviation was 20.2%.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425662 TI - The effects of differing insulin levels on the hormonal and metabolic response to equivalent hypoglycemia in normal humans. AB - The aim of this study was to determine if differing concentrations of insulin can modify the counterregulatory response to equivalent hypoglycemia in normal humans. Experiments were conducted in 9 normal, lean men, who had fasted overnight. Insulin was infused in two separate, randomized protocols so that steady-state levels of 486 +/- 33 (low) and 3056 +/- 236 pM (high) were obtained. Glucose was infused during both protocols to ensure that the rate of fall of plasma glucose (0.07 mM/min) and hypoglycemic plateau (2.8 +/- 0.1 mM) were similar. Despite similar plasma glucose levels, EPI (8.7 +/- 0.7 vs. 5.5 +/- 0.7 nM), NE (3.3 +/- 0.3 vs. 2.3 +/- 0.2 nM), and cortisol (811 +/- 36 vs. 611 +/- 72 nM) significantly increased during high compared with low insulin infusion, respectively (P < 0.05). Glucagon, growth hormone, and pancreatic polypeptide levels increased briskly and significantly but were not different during the two insulin infusions. HGP rose significantly from 12.1 +/- 0.3 to 18.1 +/- 1.1 mumol.kg-1 x min-1 in response to the high insulin level (P < 0.05) but remained unchanged (12.1 +/- 0.4 and 11.7 +/- 1.4 mumol.kg-1 x min-1) in the presence of th low insulin level. GRa increased significantly during high insulin levels (3.4 +/- 0.3 to 4.8 +/- 0.7 mumol.kg-1 x min-1, P < 0.05) but remained at a basal rate (3.0 +/- 0.3 to 2.7 +/- 0.6 mumol.kg-1 x min-1) in the presence of low insulin levels. sBP and heart rate increased more during high insulin infusion (18 +/- 5 vs. 6 +/- 5 mmHg and 18 +/- 4 vs. 7 +/- 2 beats/min, respectively, P < 0.05). In summary, the 6-fold higher insulin level resulted in significantly greater increases in catecholamine and cortisol secretion, HGP, lipolysis, heart rate, and sBP despite equivalent hypoglycemia. We conclude that at moderate hypoglycemia, high doses of insulin can augment certain aspects of the counterregulatory response in normal humans. PMID- 8425663 TI - Insulin resistance in aging is related to abdominal obesity. AB - Studies have shown that insulin resistance increases with age, independent of changes in total adiposity. However, there is growing evidence that the development of insulin resistance may be more closely related to abdominal adiposity. To evaluate the independent effects of aging and regional and total adiposity on insulin resistance, we performed hyperinsulinemic euglycemic clamps on 17 young (21-33 yr) and 67 older (60-72 yr) men and women. We assessed FFM and total and regional adiposity by hydrodensitometry and anthropometry. Insulin stimulated GDRs at a plasma insulin concentration of approximately 450 pM averaged 45.6 +/- 3.3 mumol.kg FFM-1 x min-1 (mean +/- SE) in the young subjects, 45.6 +/- 10.0 mumol.kg FFM-1 x min-1 in 24 older subjects who were insulin sensitive, and 23.9 +/- 11.7 mumol.kg FFM-1 x min-1 in 43 older subjects who were insulin resistant. Few significant differences were apparent in skin-fold and circumference measurements between young and insulin-sensitive older subjects, but measurements at most central body sites were significantly larger in the insulin-resistant older subjects. Waist girth accounted for > 40% of the variance in insulin action, whereas age explained only 10-20% of the total variance and < 2% of the variance when the effects of waist circumference were statistically controlled. These results suggest that insulin resistance is more closely associated with abdominal adiposity than with age. PMID- 8425664 TI - No effect of naloxone on ventilatory response to progressive hypercapnia in IDDM patients. AB - The ventilatory response to hyperoxic progressive hypercapnia was examined by comparing 3 test groups: 7 diabetic patients with AN, 8 diabetic patients without AN, and 8 normal control subjects. In each group, a significant linear correlation was found between PaCO2 and VE. The slopes of the regression curves relating PaCO2 to VE were significantly steeper in the healthy control subjects and diabetic patients without AN than in those with AN (P < 0.01). We conclude that the ventilatory response to progressive hypercapnia is reduced in diabetic patients with AN. By analyzing the power spectrum and the amplitude behavior of the diaphragmatic EMG (calculated from the fc and RMS, respectively), we could exclude a disturbance of neural descending pathways and respiratory muscle dysfunction as possible causal mechanisms for the impaired ventilatory response to increasing CO2. By using lung function analysis, causal factors such as alterations in respiratory system mechanics also could be excluded. As diabetes is known to affect the endogenous opioid system, which, in turn, affects the ventilatory response to CO2, naloxone, as a specific opioid antagonist, was administered in all 3 test groups. Naloxone produced a significant increase of ventilatory response to hypercapnia in the healthy control subjects (P < 0.01), but produced no effect in either of the diabetic groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425665 TI - Early exposure to cow's milk and solid foods in infancy, genetic predisposition, and risk of IDDM. AB - Using a case-control study design, we examined the hypothesis that early exposure to cow's milk and solid foods increased the risk of IDDM. An infant diet history was collected from 164 IDDM subjects from the Colorado IDDM Registry with a mean birth year of 1973, and 145 nondiabetic population control subjects who were frequency matched to diabetic subjects on age, sex, and ethnicity. Early exposure was defined as exposure occurring before 3 mo of age. After controlling for ethnicity, birth order, and family income, more diabetic subjects were exposed early to cow's milk (OR 4.5, 95% CI 0.9-21.4) and solid foods (OR 2.5, CI 1.4 4.3) than control subjects. To examine this association while accounting for the genetic susceptibility to IDDM, we defined individuals as high and low risk by an HLA-DQB1 molecular marker. Early exposure to cow's milk was not associated with elevated risk for IDDM in low-risk individuals. Relative to unexposed low-risk individuals, early exposure to cow's milk was strongly associated in individuals with a high risK marker (OR 11.3, CI 1.2-102.0). Similar findings were observed for early exposure to solid foods. These data indicate that early exposure to cow's milk and solid foods may be associated with increased risk of IDDM. The inclusion of HLA-encoded risk in the analyses demonstrates the combined effect of genetic and environmental factors. PMID- 8425666 TI - Effects of epinephrine on insulin secretion and action in humans. Interaction with aging. AB - This study was designed to define the effects on glucose metabolism of small increases of plasma EPI, comparable to increases observed during physiological sympathoadrenal activation. This study was also designed to determine the effects of EPI on glucose metabolism in older adults, in whom changes in adrenergic responsiveness of several tissues were described. Tolbutamide-boosted IVGTTs were performed during intravenous infusions of saline (control) or EPI at 2.7, 5.5, and 10.9 mmol/min to achieve physiological levels of EPI in 7 young subjects (19 26 yr of age) and 7 old subjects (62-75 yr of age), all with a normal screening OGTT. IVGTT results were analyzed to determine the AIR and with the minimal model method of Bergman to determine SI and SG. A significant fall was observed in AIR, SI, and SG for all subjects, even with the lowest dose of EPI, which resulted in only a two- to threefold increase in plasma EPI. Older subjects had a delayed recovery from hyperglycemia during the EPI infusions, although we detected no significant differences between the young and old subjects in the ability of EPI to alter either acute phase insulin secretion or insulin action. In contrast, the impairment of SG by EPI appeared to be greater in the elderly. We conclude that small increases of plasma EPI can significantly affect determinants of glucose tolerance in both young and old people. PMID- 8425667 TI - A new model for the study of mild diabetes during pregnancy. Syngeneic islet transplanted STZ-induced diabetic rats. AB - Diabetes during pregnancy carries short- and long-term consequences for the offspring. Improved obstetrical and diabetic care has resulted in decreased morbidity and mortality in the neonate of the diabetic mother. Mild hyperglycemia is still found in both IDDM pregnant women and women with GDM. The long-term consequences of exposure to mild hyperglycemia in utero remain to be determined. In an effort to develop an appropriate animal model of mild diabetes during pregnancy, we mated female STZ-induced diabetic rats previously transplanted with specific numbers of islets of Langerhans (2500, 1000, 700, or 500 islets). Diabetic and nondiabetic sham-transplanted control groups also were studied. During pregnancy, the plasma glucose levels in the diabetic rats and the group receiving 500 islets (26.5 +/- 1.1 and 10.0 +/- 0.8 mM, respectively) were significantly greater than in control animals (5.4 +/- 0.5 mM, P < 0.0001). The mean glucose levels in rats receiving 700 or 1000 transplanted islets (6.8 +/- 0.2 and 6.5 +/- 0.2 mM) also were significantly greater than in control animals (P < 0.001). No difference was evident between control rats and the group receiving 2500 islets (5.8 +/- 0.2 mM). No gross congenital abnormalities were apparent in the offspring. The pup plasma glucose was significantly greater in the offspring of dams receiving either none (diabetic) or 500 islets (10.6 +/- 0.7 and 11.1 +/- 1.1 mM, respectively) compared with the offspring of nondiabetic control dams (4.4 +/- 0.3 mM, P < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425669 TI - Influence of islet amyloid polypeptide and the 8-37 fragment of islet amyloid polypeptide on insulin release from perifused rat islets. AB - IAPP, or amylin, is a 37-amino acid peptide that is co-secreted with insulin from the pancreatic beta-cells. We have determined the effects of IAPP and the antagonist 8-37 fragment of IAPP on the secretion of insulin from isolated rat islets studied in a perifusion system. Insulin secretion was stimulated by 8 mM glucose and 0.2 microM carbachol. IAPP at 10(-7) M reduced insulin release by 32% from 7.1 (95% Cl 5.8-8.6) to 4.8 (3.0-7.5) fmol.min-1 x islet-1 (P = 0.046, n = 7). IAPP at 1.5 x 10(-6) M reduced insulin release by 62% from 6.5 (3.4-12.3) to 2.5 (1.4-4.4) fmol.min-1 x islet-1 (P = 0.001, n = 6). IAPP at 10(-5) M decreased insulin release by 70% (P < 0.001, n = 6). When IAPP (8-37) at 10(-5) M was added to IAPP at 1.5 x 10(-6) M, there was only a 22% reduction of insulin release (P = 0.06, n = 6) compared with control chambers with no peptide added. This reduction was less (P = 0.002) than observed with IAPP (1.5 x 10(-6) M) alone. IAPP (8-37) at 4 x 10(-5) M in the absence of exogenously added IAPP increased insulin secretion by 48% (P = 0.01, n = 6), but IAPP (8-37) at 10(-5) M did not alter insulin secretion. These findings demonstrate that IAPP decreases insulin secretion from islet beta-cells, an effect that can be antagonized by the 8-37 fragment of IAPP.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425668 TI - Effect of STZ administration on islet isograft and allograft survival in NOD mice. AB - We examined the effect of a single STZ administration on subsequent islet isograft and allograft survival in NOD mice. Young prediabetic NOD mice were rendered diabetic by STZ administration and transplanted with islet isografts 8 11 days later. The earliest loss of islet function occurred on postgraft day 49. In sharp contrast, 15 of 16 isografts in spontaneously diabetic mice were destroyed within 17 days postgrafting. A comparison of the age of islet isograft destruction in STZ-induced diabetic NOD mice with the course of diabetes development in the NOD mouse colony clearly showed that STZ administration at the prediabetic stage led to a significant delay of diabetes onset in isografts. When STZ was given to overtly diabetic NOD mice, both islet isografts and allografts survived significantly longer than those in untreated, spontaneously diabetic NOD mice. However, the degree of prolongation induced by STZ was much smaller compared with that induced by ALS, a potent immunosuppressive reagent. In vitro mixed lymphocyte culture experiments showed that spleen cells of mice given STZ exhibited time-dependent reduction of their alloantigen reactivities. These results demonstrate that STZ, which is commonly used to induce diabetes in various experimental animals, also possesses an immunosuppressive property, although it is relatively weak compared with ALS. PMID- 8425670 TI - Effects of ponalrestat, an aldose reductase inhibitor, on neutrophil killing of Escherichia coli and autonomic function in patients with diabetes mellitus. AB - In diabetic subjects, polyol pathway activity might inhibit neutrophil function and cause nerve damage. The effects of ponalrestat, an aldose reductase inhibitor, were assessed on neutrophil intracellular killing of Escherichia coli and on autonomic function in diabetic subjects in a randomized double-blind, placebo-controlled, crossover trial. We studied 31 diabetic subjects with autonomic dysfunction and 21 age- and sex-matched control subjects. During two 12 wk treatment periods, the diabetic subjects took either 600 mg of ponalrestat or matching placebo once daily. Neutrophil killing of E. coli was measured by a microbiological assay technique. Kmax by neutrophils from the diabetic subjects was lower than in the control group (Kmax of diabetic subjects 54.5 +/- 26.4 vs. control subjects 67.3 +/- 16.3, P = 0.045). Ponalrestat significantly increased bacterial killing in the diabetic subjects (Kmax of ponalrestat 75.1 +/- 16.5 vs. placebo 58.2 +/- 20.8, P = 0.003) so that there was no longer any significant difference in Kmax between the control subjects and the diabetic subjects on active treatment. Ponalrestat had no significant effect on a range of standard cardiovascular autonomic nerve function tests. We conclude that neutrophil killing of E. coli is impaired in diabetic subjects with autonomic dysfunction. This is restored to normal by ponalrestat. PMID- 8425671 TI - Severe hypoglycemia and intelligence in adult patients with insulin-treated diabetes. AB - The IQ scores (WAIS-R) of 100 patients with insulin-treated diabetes (aged 25-52 yr) were compared with those of 100 healthy control subjects who were matched to the diabetic patients for sex, age, education, and social class. The diabetic group had lower WAIS-R performance and verbal IQ scores than the control group (P = 0.017 and P = 0.033, respectively) after controlling for premorbid IQ. The extent of the difference was modest, representing approximately 33% of an SD in IQ. When frequency of severe hypoglycemia was controlled for the difference in performance IQ between the diabetic patient group and the control group was abolished, whereas the difference between the groups in verbal IQ persisted. It is hypothesised that cumulative severe hypoglycemia might be the major factor in the slight performance IQ differences between diabetic patients and control subjects. The origin of the verbal IQ differences, although obscure, might be related to the social impact of the disorder. PMID- 8425672 TI - Immunochemical detection of advanced glycation end products in lens crystallins from streptozocin-induced diabetic rat. AB - To reassess the significance of AGEs in cataract formation in diabetic animals, we measured amounts of AGEs in lens crystallins from STZ-induced diabetic animals with a newly developed ELISA. Lenses were removed at 5 and 20 wk after STZ injection. In 20-wk diabetic rats, all lenses were cataractous but not in control rats. In 20-wk diabetic compared with control rats, significant increases were observed in AGEs (172.3 +/- 18.3 vs. 14.3 +/- 1.7 AU, P < 0.01) and fluorescence (2.04 +/- 0.22 vs. 1.27 +/- 0.10 AU, P < 0.05). The amounts of AGEs in lens crystallins, measured by the ELISA, were > 12-fold higher in diabetic rats. In agreement with earlier studies, we found that fluorescence in lens crystallins increased by 61% in diabetic rats. In 5-wk diabetic rats, all lenses were noncataractous. In 5-wk diabetic compared with control rats, significant increases were observed in AGEs (84.1 +/- 7.7 vs. 9.4 +/- 1.5 AU, P < 0.01) and fluorescence (1.45 +/- 0.06 vs. 1.05 +/- 0.06 AU, P < 0.01). Analysis of the AGE content by ELISA showed that accumulation of AGEs in diabetic lens crystallins does markedly occur with time, and a large amount of AGEs exists in the diabetic (cataractous) lens crystallins. The disproportionate elevation of AGEs, measured by the ELISA, compared with fluorescence suggests that the actual levels of AGEs in cataractous lens crystallins from diabetic animals are higher than previously anticipated, and nonfluorescent AGEs may exist in diabetic lens crystallins.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425673 TI - Insulin stimulates production and secretion of endothelin from bovine endothelial cells. AB - Endothelin, a vasoconstrictor peptide secreted from endothelial cells, has been thought to play a role in various forms of vascular disease. Diabetes mellitus is well known for its association with accelerated atherosclerosis and microvascular damage. Although the basis for the vessel insult is multifactorial, hyperinsulinemia is thought to contribute by an unknown mechanism. In this study, we sought to determine whether insulin stimulates the production and secretion of ET-1 as a possible basis for the association of hyperinsulinemia and vascular disease. We demonstrated that insulin significantly stimulates the gene expression and secretion of ET-1 from cultured BAEC, and that insulin increases ET-1 mRNA expressed in BBCEC. Insulin caused a maximal twofold inducement above control ET-1 mRNA expression in a dose-related fashion in BAEC. The increased mRNA resulted from increased transcription, as determined by nuclear run-off studies. Increased ET-1 mRNA was seen after 4 h of incubation with insulin: the peak occurred at 6-8 h and persisted for 24 h. Insulin caused as much as a fourfold stimulation of ET-1 secretion from BAEC in a dose-related fashion, including a twofold increase at a physiological concentration (10(-9) M): The increase began at 1 h of incubation and continued for the entire 24-h incubation period. The insulin-induced increases in both ET-1 mRNA and ET-1 protein secretion were significantly attenuated by genistein, a tyrosine kinase inhibitor. This stimulation probably occurred through the insulin receptor, because IGF-1 had no effect on ET-1 gene expression or secretion from these cells.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425674 TI - Antibodies to glutamic acid decarboxylase reveal latent autoimmune diabetes mellitus in adults with a non-insulin-dependent onset of disease. AB - The classification of adults with diabetes mellitus can be invalidated by patients who initially present as NIDDM but who later become frankly insulin dependent. In some of these, the pathogenesis could be similar to that in IDDM, namely autoimmune destruction of the pancreatic beta-cells. We studied 102 patients > 35 yr of age at diabetes onset who had initially been nonketotic and non-insulin-dependent for > or = 6 mo. They were classified according to glucagon stimulated C-peptide levels into an insulin-deficient group (n = 33) and a non insulin-deficient group (n = 69). We measured antibodies to GAD, islet cell cytoplasm, thyroid antigens, and gastric parietal cells in both groups. Anti-GAD was significantly higher in the insulin deficient group, 76% (25 of 33), than in the non-insulin deficient group, 12% (8 of 69), and this difference was substantially greater than that shown for ICAs. Thus, in a proportion of adults who present with NIDDM, a slowly evolving autoimmune insulitis can be revealed by testing for anti-GAD. This could have important connotations not only for early intervention, but also for the correct classification of diabetes. PMID- 8425675 TI - Development and application of endoscopy. PMID- 8425676 TI - Ultrastructure of interstitial cells of Cajal in circular muscle of human small intestine. AB - BACKGROUND: Interstitial cells of Cajal (ICC) may be important regulatory cells in gut muscle layers. This study examined ICC within the circular muscle of human small intestine. METHODS: Surgically resected, uninvolved intestine was studied by light microscopy and electron microscopy. RESULTS: Muscle lamellae were separated by main septa in continuity with submucosa. Smooth muscle cells ran radially in the septa. Two types of ICC were distinguished. One ICC type had abundant intermediate filaments and smooth cisternae and a discontinuous basal lamina. This ICC type was present in the septa and in the outer third of the circular lamellae. The other ICC type had a complete basal lamina and conspicuous caveolae. This ICC type was observed only in the inner third of the circular lamellae. Both ICC types were close to nerves, but only the latter type formed gap junctions with one another and with muscle cells. Junctions between the two ICC types were not observed. CONCLUSIONS: The arrangement suggests that ICC and radially oriented muscle cells participate in electrical and mechanical coordination of the circular muscle layer of human small intestine. PMID- 8425677 TI - An in-depth study of Crohn's disease in two French families. AB - BACKGROUND: Two French families were investigated. In the first a husband, wife, and 4 children had Crohn's disease; in the second 7 of 11 children had the disease. There was no history of Crohn's disease in antecedent generations and no linkage to HLA haplotypes. METHODS: Methods included family interviews; review of medical records, radiographs, and pathology slides; serology; selective stool culture; enzyme-linked immunosorbent assay for fecal viral detection; and immunocytochemistry. RESULTS: In both families multiple cases occurred among siblings in 7-13-month periods. There appeared to be a 4-8-year recurrence of new disease in both families. Radiographs showed a remarkable similarity in the pattern of disease, confined to distal ileum and cecum, in the members of family 1. Examination for pathology showed granulomas in all 8 patients for whom tissues were available. Acid-fast organisms or Campylobacter-like organisms were not found in tissue sections, and immunocytochemistry was negative for mycobacteria and Yersinia. Stool cultures were negative for mycobacteria, Yersinia, and Mycoplasma. Torovirus and coronavirus antigens were not found in stool. Serology was negative for antibodies to Brucella, Yersinia, influenza, and three enteropathogenic viruses of animals. CONCLUSIONS: The circumstances and data suggest that an infectious microorganism is responsible for these clusterings of Crohn's disease. PMID- 8425678 TI - In vitro changes in the properties of rabbit colonic muscularis mucosae in colitis. AB - BACKGROUND: The muscularis mucosae is the muscle layer closet to the site of elevated inflammatory mediator production in inflammatory bowel disease. Thus, it is the first muscle layer subject to their influence. METHODS: Using a rabbit trinitrobenzene sulfonic acid model of colitis, changes in the properties of the muscularis mucosae resulting from the inflammatory process were studied in vitro. RESULTS: Animals developed a mild colitis-like inflammation that was confined to the epithelium, lamina propria, and submucosa. Colitic muscularis mucosae contractile responses to leukotriene D4 and prostaglandins E2 and F2 alpha were significantly attenuated relative to the maximum tissue response to acetylcholine, whereas responses to histamine, substance P, and vasoactive intestinal polypeptide were unchanged. In addition, the stress-generating capacity of the colitic muscularis mucosae was compromised in a stimulus independent manner and passive tension increased relative to active tension. CONCLUSIONS: The muscularis mucosae undergoes two significant alterations in colitis: (a) a selective desensitization to the effects of arachidonic acid metabolites and (b) an impairment of its excitation-contraction coupling mechanism. A loss of the ability of the muscularis mucosae to cause mucosal movement and alter luminal surface area may be an important early stage in the pathophysiology of inflammatory bowel disease. PMID- 8425679 TI - Central action of interleukin 1 beta on intestinal motility in rats: mediation by two mechanisms. AB - BACKGROUND: Interleukin 1 (IL-1) can influence gut functions by inhibiting gastric acid secretion. This study was performed to investigate the effects of IL 1 on intestinal motility and the mechanisms involved. METHODS: The effects of IL 1 beta were determined by electromyography in conscious rats with implanted electrodes and a permanent catheter in a lateral brain ventricle. RESULTS: Intracerebroventricular IL-1 beta (15 ng) administered to fed rats immediately stimulated cecocolonic spike bursts and caused a migrating myoelectric complex pattern after a delay in the small intestine. Tenfold higher doses of peripherally administered IL-1 beta did not promote similar reactions. The IL-1 antagonist reduced the small intestinal effect of IL-1 beta and blocked the cecocolonic stimulation. Indomethacin and SC 19220 reduced the small intestinal effects but did not antagonize the increase in cecocolonic contractions. In contrast, alpha-helical CRF9-41 blocked the increase of cecocolonic contractions but did not antagonize the IL-1 beta-induced effects on the small intestine. CONCLUSION: IL-1 beta's effects on intestinal motility can be mainly ascribed to a central action. The cecocolonic stimulation may be mediated by brain corticotropin-releasing factor, whereas the small intestinal effects involve a prostaglandin mediation. PMID- 8425680 TI - Major histocompatibility antigen-restricted cytotoxicity in inflammatory bowel disease. AB - BACKGROUND: The role of cytotoxicity mediated by peripheral blood mononuclear cells for colonic epithelial cells in inflammatory bowel disease (IBD) is still controversial. To clarify it, we studied major histocompatibility antigen (MHC) restricted T cell-mediated cytotoxicity (CTL). METHODS: Cytotoxicity was measured by 51Cr release from colonic cells after the 6-hour incubation with peripheral blood mononuclear cells in 11 IBD patients (6 with Crohn's disease and 5 with ulcerative colitis). RESULTS: CTL activity (E/T ratio = 200:1 or 100:1) for autologous target cells was significantly increased (22%-40%) in 5 of 6 CD and 4 of 5 UC patients (22%-64%) compared with that for allogeneic target cells. The increase in CTL activity was mainly inhibited by anti-MHC class I and CD8 monoclonal antibodies (50 micrograms/mL), while it was partially inhibited by anti-MHC class II or CD4 antibodies in some patients. Complement-mediated depletion of CD2+ cells also significantly decreased CTL activity. CONCLUSIONS: The results indicate that MHC-restricted T cell cytotoxicity may play a role in mucosal damage in some patients of IBD. PMID- 8425681 TI - Impaired hydrogen metabolism in pneumatosis cystoides intestinalis. AB - BACKGROUND: Pneumatosis cystoides intestinalis (PCI) is characterized by high levels of breath hydrogen. Clinical features of PCI may be due to abnormal H2 metabolism. METHODS: Breath levels of H2 and CH4 were measured in 3 patients and total gas in 2 patients with PCI on a polysaccharide-free (basal) diet and after administration of 15 g of lactulose. Metabolic activities and counts of methanogenic (MB) and sulfate-reducing (SRB) bacteria were measured in feces. Ten volunteers were also studied. RESULTS: Total H2 levels in patients were 383-420 mL/day on the basal diet and 1430-1730 mL/day after lactulose administration compared with 35 +/- 6 mL/day and 262 +/- 65 mL/day, respectively, in controls. Basal breath H2 levels in controls were 27 +/- 6 vs. 214 +/- 27 mL/day in patients and after lactulose ingestion, 115 +/- 18 vs. 370 +/- 72 mL/day. Four controls were methanogenic and had high fecal MB counts. The other controls had high SRB counts and sulfate reduction rates. All patients were nonmethanogenic and had low sulfate reduction rates. CONCLUSIONS: Patients with PCI excrete more H2 than controls. In normal subjects, H2 is consumed by MB or SRB; the activity of these bacteria is virtually absent in PCI. This may explain the gas accumulation in these patients. PMID- 8425682 TI - Nitric oxide mediates inhibitory nerve input in human and canine jejunum. AB - BACKGROUND: Nitric oxide (NO) may be an inhibitory neurotransmitter in the intestinal muscle. The present study examined its role in human and canine jejunum. METHODS: Mechanical and intracellular electrical activity were recorded simultaneously from the circular muscle layer. RESULTS: In the human jejunum, nerve stimulation inhibited mechanical activity and evoked an inhibitory junction potential that consisted of an initial fast hyperpolarization followed by a late sustained hyperpolarization. NO inhibited mechanical activity and evoked a dose dependent hyperpolarization that mimicked the late hyperpolarization. In the canine jejunum, nerve stimulation inhibited mechanical activity and evoked an inhibitory junction potential that consisted of only a fast hyperpolarization. NG Monomethyl-L-arginine and NG-nitro-L-arginine attenuated nerve-mediated inhibition of mechanical activity in both species. However, the effect of the synthase inhibitors on inhibitory junction potentials differed in the two species. In canine jejunum, both inhibitors reduced the amplitude of the initial fast hyperpolarization. In human jejunum, both inhibitors reduced only the late sustained hyperpolarization. CONCLUSIONS: NO mediates neural inhibition in circular muscle of both human and canine jejunum through different mechanisms. PMID- 8425683 TI - Effects of cigarette smoking on solid and liquid intragastric distribution and gastric emptying. AB - BACKGROUND: The acute effects of cigarette smoking on gastric emptying are controversial, whereas its effects on the intragastric distribution of solids and liquids are not established. METHODS: Dual isotope gastric scintigraphy was performed in 15 habitual smokers (studied twice, either sham smoking or actively smoking) and in 15 age- and sex-matched nonsmokers. RESULTS: Acute smoking was associated with an increased prevalence of episodes of retrograde intragastric movement of solids (3 of 15 sham subjects vs. 12 of 15 actively smoking subjects; P < 0.01) and of liquids (0 of 15 vs. 7 of 15; P < 0.01) from distal to proximal stomach. Fundal half-emptying time (T1/2) for liquids was also prolonged by smoking (43 +/- 19 minutes sham vs. 125 +/- 216 minutes active; P < 0.05). Acute smoking delayed solid lag time (13 +/- 6 minutes sham vs. 32 +/- 18 active; P < 0.05) and liquid T1/2 (46 +/- 21 vs. 90 +/- 50 minutes; P < 0.05). In the nonsmokers, such episodes of proximal intragastric redistribution did not occur, and intragastric and overall emptying parameters did not differ significantly from those of habitual sham smokers. CONCLUSIONS: Acute cigarette smoking produces excessive antrofundal redistribution of both solid and liquid contents and delays solid and liquid gastric emptying. PMID- 8425684 TI - Effects of a new sigma ligand, JO 1784, on cysteamine ulcers and duodenal alkaline secretion in rats. AB - BACKGROUND: The ulceroprotective effects of JO 1784 [(+)-N-cyclopropyl-methyl-N methyl-1,4-diphenyl-1-ethyl-but-3-en-1-yl amine, hydrochloride], a new specific and highly selective sigma ligand, were examined in rats. METHODS: Different models of gastric ulcers (4-hour restraint stress, aspirin, ethanol, and taurocholate) and cysteamine-induced duodenal ulcers were used. The gastric acid secretion (4-hour Shay rat preparation) and the duodenal bicarbonate secretion were also studied. RESULTS: JO 1784 elicited a potent protection against duodenal ulcers but had a weaker protective effect on any of the gastric ulceration models tested. It displayed no gastric antisecretory activity but induced a dose dependent stimulation of duodenal bicarbonate secretion. Haloperidol, hexamethonium, tetrodotoxin, bivagotomy (but not atropine), and the intravenous but not intracerebroventricular administration of devazepide, a cholecystokinin A antagonist, inhibited the stimulatory effect of JO 1784. CONCLUSION: These results show that JO 1784, a selective sigma ligand, is a potent protector of the duodenal mucosa. This activity may be related to its stimulating effect on bicarbonate secretion, which is driven through a complex nervous mechanism involving muscarinic synapses, vagal afferent fibers, and peripheral cholecystokinin receptors. This drug might open a new specific way in the treatment of duodenal ulcers. PMID- 8425685 TI - Oral mesalamine (Pentasa) as maintenance treatment in Crohn's disease: a multicenter placebo-controlled study. The Groupe d'Etudes Therapeutiques des Affections Inflammatoires Digestives (GETAID) AB - BACKGROUND: Mesalamine provides a new therapeutic approach in treating Crohn's disease. METHODS: To assess the efficacy and safety of slow-release mesalamine (Pentasa; Ferring AS, Vanlose, Denmark) in maintaining remission in Crohn's disease, 161 patients with inactive disease were randomized to receive either Pentasa (2 g/day) or placebo in a 2-year double-blind, multicenter trial. Two strata were defined according to the duration of their remission: < 3 months (n = 64) or 3-24 months (n = 97), presumed to be high and a low relapse risk strata, respectively. RESULTS: The probability of relapse was higher in the short remission placebo group than in the three other groups (P < 0.003), showing there was a significant benefit from Pentasa in the high relapse risk stratum. In this stratum, the 2-year ongoing remission rate was of 29% +/- 9% and 45% +/- 11% (mean +/- SD) in the placebo and Pentasa groups, respectively. The incidences of side effects were similar in both groups. CONCLUSIONS: Pentasa (2 g/day for 2 years) is a safe and effective maintenance treatment for Crohn's disease when given within 3 months of achieving remission. PMID- 8425686 TI - Prostanoids inhibit intestinal NaCl absorption in experimental porcine cryptosporidiosis. AB - BACKGROUND: Recent studies of piglet cryptosporidiosis have shown that impaired Na(+)-coupled glucose absorption is associated with a loss of two thirds of the villous absorptive surface and an inflammatory infiltration of the lamina propria. Because inflammatory cells release eicosanoids that may alter electrolyte transport, the present study examined the role of prostanoids on NaCl transport. METHODS: Ileal mucosa was stripped of its muscle layers and mounted in Ussing chambers in the presence or absence of indomethacin. Adjacent tissue was also frozen for subsequent extraction and radioimmunoassay of prostaglandin E2 (PGE2). RESULTS: Results showed that net Na+ absorption is inhibited and net Cl- secretion is induced in infected piglets. Indomethacin restored net Na+ and Cl- absorption to control levels and exogenous PGE reversed this effect. Radioimmunoassay of tissue extracts showed that PGE2 increased from 56.7 +/- 9.6 ng/cm2 in control to 134 +/- 16.8 ng/cm2 in infected ileum (P < 0.01). CONCLUSIONS: These data indicate that in addition to the Na-glucose malabsorption arising from structural damage, part of the diarrhea of these infected animals must be attributed to local prostanoid production. PMID- 8425687 TI - Immune activation genes in inflammatory bowel disease. AB - BACKGROUND: Inflammatory bowel disease is associated with enhanced activation of T cells, but the genes responsible for this state are not well characterized. METHODS: T-cell activation genes were studied in peripheral blood and intestinal mucosal mononuclear cells of control, Crohn's disease, and ulcerative colitis patients. RESULTS: In all groups the expression of interleukin-2 (IL-2), IL-2 receptor alpha (IL-2R alpha), and IL-2R beta messenger RNA (mRNA) was significantly higher in intestinal than circulating cells, and it correlated well with protein levels. Both IL-2R alpha and IL-2R beta mRNA were abundant in mucosal cells, suggesting that a substantial number of them displays high affinity IL-2R. This would explain why intestinal cells proliferate more, express more IL-2 transcripts, and secrete more IL-2 than peripheral cells. Inflammatory bowel disease cells produced similar or higher IL-2R alpha and IL-2R beta mRNA than controls but generated significantly lower IL-2 mRNA. Thus, the reported defect of IL-2 activity in Crohn's disease and ulcerative colitis is probably related to decreased IL-2 transcription. Crohn's disease intestinal cells had the highest expression of IL-2R gene products. This provides a mechanism for their increased response to IL-2 and supports claims that elevated soluble IL-2R alpha serum levels reflect gut T-cell hyperactivity in this disease. CONCLUSIONS: These findings underscore the importance of T cells in mucosal immunity and indicate that abnormal T-cell activation is intimately associated to the pathogenesis of inflammatory bowel disease. PMID- 8425688 TI - Ileal proglucagon gene expression in the rat: characterization in intestinal adaptation using in situ hybridization. AB - BACKGROUND: Proglucagon-derived peptides are potential mediators of the adaptive response of the terminal ileum to massive small bowel resection. Ileal proglucagon messenger RNA (mRNA) levels increase during ileal adaptation. The present study explored the cellular basis of this response. METHODS: Sections of control ileum, ileum 4 days after resection, and pancreas were analyzed by in situ hybridization with 35S-labeled complementary RNA (cRNA) probes. RESULTS: Both the proglucagon and the peptide YY cRNA probes hybridized to discrete cells in the ileal mucosa, the disposition of which corresponds to that reported for intestinal L cells. Four days after resection there was a marked increase in the intensity of the signal for both probes without an increase in cell number. Insulin and histone H3 probes were used as controls to confirm the specificity of the hybridization seen with the L-cell specific, proglucagon, and peptide YY probes. CONCLUSIONS: The increase in proglucagon mRNA levels after massive small bowel resection is caused by an increase in the cellular content. The parallel increase in PYY mRNA levels implies an L cell--rather than a proglucagon gene- specific response. PMID- 8425689 TI - The effect of enterohemorrhagic Escherichia coli O157:H7 on intestinal structure and solute transport in rabbits. AB - BACKGROUND: The effect of enterohemorrhagic Escherichia coli O157:H7 infection on intestinal morphology and solute transport was examined. METHODS: New Zealand white rabbits, aged 10 days, were infected with E. coli strain EDL933 (O157:H7 containing the 60-megadalton plasmid-encoding adhesion factors VT1 and VT2) and compared with controls. Small and large intestinal histology and solute transport were studied 5 days after inoculation. Ion transport in the distal colon was also examined in animals infected with different strains encoding a combination of pathogenic factors. RESULTS: Infection with EDL933 induced diarrhea and mucosal disease in the colon, inhibited colonic Na+ absorption, and stimulated of Cl- secretion, but had no impact on the small intestine. Infection with strains A7785 C3A (O157:H7, plasmid-, VT1+, VT2+) and 85-170 (O157:H7, plasmid+, VT-) induced similar transport changes to EDL933. C600/1 (E. coli K-12, plasmid+, VT1+) decreased Na+ and Cl- absorption only. CONCLUSIONS: Abnormalities of colonic structure and ion transport could account for diarrhea production, but pathogenic factors other than the 60-megadalton plasmid-encoding adhesion factor and verotoxins appear to be involved in enterohemorrhagic E. coli infection. PMID- 8425690 TI - Ventromedial hypothalamic lesions increase gastrointestinal DNA synthesis through vagus nerve in rats. AB - BACKGROUND: It was recently reported that ventromedial hypothalamic (VMH) lesions produced an increase in gastrointestinal DNA content in rats. In the present study, the mechanism of this alteration was examined. METHODS: The DNA content and synthesis after VMH lesioning in rat gastrointestinal tracts were determined. RESULTS: Total content of DNA in stomach and small intestine began to increase at 3 days and continued to increase for 7 days, whereas DNA content in the large intestine began to increase at 3 days and maintained the same level until 7 days after VMH lesioning. DNA synthesis of these organs increased and reached maximum at 3 days and then decreased to the initial level 7 days following the lesions. This increase in DNA content and synthesis in these organs was largely inhibited by bilateral subdiaphragmatic vagotomy or the administration of atropine, a cholinergic blocker, but not by the administration of anti-insulin antibody. CONCLUSIONS: VMH lesions induce cell proliferation in the rat gastrointestinal tract by the firing of vagus nerve activity mainly through the cholinergic receptor mechanism. PMID- 8425691 TI - Maturation of human fetal stomach in organ culture. AB - BACKGROUND: This investigation was undertaken to establish a serum-free organ culture technique allowing for the morphological and physiological maintenance of human fetal stomach in vitro. METHODS: Explants from gastric corpuses (12-17 weeks of gestation) were cultured in serum-free medium for periods of up to 15 days. RESULTS: After 15 days of culture, surface mucous cells were more mature, gastric glands were numerous and well developed, and all epithelial cell types were morphologically very well preserved. Morphometric measurements of the glands revealed an accelerated development in culture compared with that found in utero. Even though the incorporation of [3H]thymidine into total DNA decreased, the labeling indices determined by radioautography confirmed that epithelial cell proliferation was maintained especially in the pit/neck portion and at the base of the glandular compartments. A significant increase in total glycoprotein synthesis, as evaluated by the incorporation of [3H]glucosamine, was observed and correlated with the differentiation of the mucous cells. CONCLUSIONS: This investigation establishes for the first time that human gastric mucosa can be maintained up to 15 days in organ culture and that maturation of the gastric mucosa can be reproduced in chemically defined media. PMID- 8425692 TI - The central organization of the vagus nerve innervating the colon of the rat. AB - BACKGROUND: The extent to which the vagus nerve innervates the colon remains controversial. METHODS: In 29 rats the tracer cholera toxin-horseradish peroxidase was injected into the cecum, the ascending, transverse, or descending colon or the rectum. For comparison, control injections were made into the stomach. RESULTS: For all areas of colon except the rectum, brainstem motoneuronal labeling was limited to the lateral third of the dorsal motor nucleus of the vagus nerve bilaterally. In contrast, gastric injections resulted in motoneuronal labeling limited to the medial portions of the nucleus. The number of labeled motoneurons was greatest following injection of the cecum, and it significantly decreased for the more distal areas of the colon. Colonic motoneuron dendrites projected into the nucleus of the solitary tract and within the dorsal motor nucleus of the vagus nerve. Sensory afferent terminal labeling was limited to the commissural and medial subnuclei of the nucleus of the solitary tract. For the rectum, sensory and motor labeling was limited to the spinal cord. CONCLUSIONS: The distribution of labeling within the vagal complex indicates that all regions of the colon, except the rectum, are innervated by the celiac and accessory celiac branches of the vagus nerve. PMID- 8425693 TI - Increasing incidence of adenocarcinoma of the esophagus and esophagogastric junction. AB - BACKGROUND: The aim of this study was to determine whether the incidence of adenocarcinoma of the esophagus and esophagogastric junction in a well-defined population was higher than previously recognized. METHODS: Clinical records and original histological slides from patients residing in Olmsted County, Minnesota, were reviewed and compared with a previous study in the same population. RESULTS: The incidence of esophageal adenocarcinoma rose from 0.13 for 1935-1971 to 0.74 for 1974-1989, and the incidence of adenocarcinoma of the esophagogastric junction rose from 0.25 to 1.34 per 100,000 person-years. Histological review of preserved surgical specimens showed associated intestinal metaplasia (Barrett's esophagus) in 2 of 2 esophageal and in 5 of 9 esophagogastric adenocarcinomas. CONCLUSIONS: The incidence of adenocarcinoma in each location increased five to sixfold compared with the earlier study. This increase could not be explained by improved diagnostic methods or classification changes. The association with Barrett's esophagus and the parallel increased incidence of cancer in each location is evidence that adenocarcinoma of the esophagus and of the esophagogastric junction are related disorders. PMID- 8425694 TI - The histological pattern and pathological involvement of the anal transition zone in patients with ulcerative colitis. AB - BACKGROUND: The aim was to determine whether the transitional epithelium (TE) of the anal transition zone (ATZ) was involved by chronic ulcerative colitis (CUC) and whether preserving the ATZ preserves the disease. METHODS: Proctocolectomy specimens from 50 CUC patients and 50 patients with rectal cancer serving as controls were stained with alcian-blue to map the ATZ, and biopsy specimens containing adjacent TE and rectal mucosa were examined. RESULTS: The mean inflammation score (0, none; 4, severe) in TE of controls was 0.4, whereas in CUC it was 0.5. However, the mean inflammation score of the rectal mucosa within the ATZ was 0.2 in controls and 2.6 in CUC (P < 0.001). Rectal columnar epithelium extended past half of the maximum length of TE in 75% of patients (65-83%; 95% CI) and was within 1 cm of the dentate line in 89% (81%-94%). CONCLUSIONS: Although the TE of the ATZ was not inflamed, the rectal mucosa within the ATZ was. Moreover, rectal mucosa traversed half of the length of the ATZ in 75% of patients and was within 1 cm of the dentate line in fully 89%. Preserving the ATZ may preserve the disease in the majority of patients with CUC. PMID- 8425695 TI - Cyclosporin A treatment in primary biliary cirrhosis: results of a long-term placebo controlled trial. AB - BACKGROUND: Effective treatment for primary biliary cirrhosis (PBC) resulting in slower progression and improved survival remains elusive. Cyclosporin A (CyA), which has been so effective in preventing human allograft rejection, has shown promise in small numbers of patients in early studies. METHODS: Three hundred forty-nine patients with PBC were randomized to receive CyA, 3 mg.kg-1.day-1, or placebo in a multicenter study with follow-up for 6 years. The end point was death or liver transplantation. RESULTS: Cox multivariate analysis showed time from entry to death or transplantation was significantly prolonged (by up to 50%) in the CyA-treated group. Liver-related mortality was also significantly lower. However, a univariate analysis of survival showed no statistical differences between the two groups. Biochemical liver indices deteriorated more slowly in the CyA-treated group, but serum creatinine concentration was elevated > 150 mumol/L in 9%, necessitating permanent discontinuation in half of these. A reduction in the dose of CyA was required in 11% because of hypertension. CONCLUSIONS: CyA has some therapeutic potential in primary biliary cirrhosis, providing blood pressure and renal function are closely monitored. PMID- 8425696 TI - Isolation of a human biliary glycoprotein inhibitor of cholesterol crystallization. AB - BACKGROUND: About 50% of populations in developed countries have bile supersaturated with cholesterol, which is a major risk factor for cholesterol gallstone formation. Despite the prevalence of supersaturated bile, only about 10% of these populations develop gallstones. The existence of a biliary protein that inhibits cholesterol crystallization was hypothesized to explain this discrepancy. This report outlines the purification and characterization of such a human biliary glycoprotein. METHODS: Chromatographic methods were used for separation and characterization. Additional steps included activity analysis by crystal growth assay, electrophoresis, and deglycosylation. RESULTS: The glycoprotein consists of a heterodimer, M(r) of 120 kilodalton, with subunits of M(r) of 63 kilodalton and 58 kilodalton. Each of the subunits is characterized by an isoelectric point of 6.6 and shows comparable inhibitory activity. Deglycosylation of the subunits show that they share a similar polypeptide backbone (M(r) of 35 kilodalton) based upon a highly similar amino acid profile. This suggests that differential subunit glycosylation alone may account for the apparent heterodimeric structure. CONCLUSIONS: No other human biliary glycoprotein has been found thus far that shows cholesterol crystal growth inhibiting activity. Thus, it may be of importance in preventing gallstone formation in healthy populations. PMID- 8425697 TI - Isolation and characterization of a cholesterol crystallization promoter from human bile. AB - BACKGROUND: Recent studies on the pathogenesis of cholesterol gallstone disease have focused on the potential importance of an imbalance between biliary proteins having either inhibitory or promoting activities on nucleation and/or growth of cholesterol crystals as the initial stage in stone formation. The current study describes the purification and partial characterization of a 42-kilodalton biliary glycoprotein that shows concentration-dependent cholesterol crystallization-promoting activity. METHODS: Chromatographic methods were used for separation and purification. Characterization steps included electrophoresis, deglycosylation, amino acid and carbohydrate analysis, and activity analysis by crystal growth assay. RESULTS: The 42-kilodalton purified glycoprotein is an extensively glycosylated (37%) monomer with an acidic isoelectric point (pl < 4.1) that is probably based on the sialic acid content of the carbohydrate moiety. Enzymatic N-deglycosylation removes the carbohydrate moiety and inactivates the promoting activity. Furthermore, enzymatic proteolysis results in both its complete structural degradation and functional inactivation. Although the glycoprotein was isolated from normal human gallbladder biles, its presence in gallstone-associated samples is clearly shown. CONCLUSIONS: This report outlines biochemical features of a human biliary glycoprotein that may be of major pathophysiological significance in gallstone disease. PMID- 8425698 TI - Inositol trisphosphate restores impaired human gallbladder motility associated with cholesterol stones. AB - BACKGROUND: Gallbladder motility is impaired in specimens with cholesterol stones but normal with pigment stones. METHODS: Muscle cells obtained from 19 human gallbladders with cholesterol stones and 11 with pigment stones were enzymatically digested and contracted with cholecystokinin octapeptide (CCK-8), acetylcholine, and KCl. RESULTS: Muscle cells from pigment stones had a greater contraction than cells from cholesterol stones. CCK-8-induced contraction was unaffected by calcium-free media but was blocked by strontium. Potassium-evoked contraction was blocked by a calcium-free media and unaffected by strontium. Inositol triphosphate (IP-3)-induced contraction was similar to the contraction caused by CCK-8 in permeable cells from pigment stones but was greater than the response to CCK-8 in cells from cholesterol stones. CONCLUSIONS: Muscle cells from gallbladders with cholesterol stones contract less than cells from gallbladders with pigment stones; CCK-8-induced contraction only uses stored calcium; and IP-3 causes contractions of equal magnitude in cells from gallbladders with cholesterol and pigment stones. These abnormalities could result from an impaired receptor activation of the mechanism for IP-3 generation and release of stored calcium. PMID- 8425699 TI - Sulindac-associated hepatic injury: analysis of 91 cases reported to the Food and Drug Administration. AB - BACKGROUND: Recent emphasis on nonsteroidal anti-inflammatory drug (NSAID) associated hepatic injury blurs differences between NSAIDs. Accordingly, examination of hepatic injury by individual NSAIDs seemed warranted. Sulindac associated hepatic injury was selected. METHODS: From 338 reports submitted to the Food and Drug Administration, 247 were considered inadequate or unconvincing for sulindac toxicity. The remaining 91 cases of reactions to the drug were analyzed. In 15 there was histological material available. RESULTS: There were four deaths, three attributed to severe hypersensitivity and one to fulminant hepatic failure. Two thirds of the cases had clinical hallmarks of hypersensitivity. The ratio of females to males was 3.5:1; 69% of the patients were over 50 years of age. Jaundice was recorded in 67% of the patients. The pattern was cholestatic in 43%, hepatocellular in 25%, mixed in 12%, and indeterminate in 20% of the patients. Eosinophilia was significantly more frequent in patients with cholestatic injury (40%) than in those with hepatocellular injury (0). CONCLUSION: Sulindac injury involves females more than males. It can lead to cholestatic or hepatocellular injury, most often because of immunological idiosyncrasy. In some patients, metabolic idiosyncrasy may be the mechanism. This study illustrates the utility of analysis of adverse reaction reports in characterizing drug-induced injury. PMID- 8425700 TI - Octreotide ameliorates vasodilatation and Na+ retention in portal hypertensive rats. AB - BACKGROUND: Whether long-term octreotide treatment given to portal hypertensive rats could prevent or ameliorate peripheral vasodilatation and thereby modify sodium retention was investigated. METHODS: Starting at the time of partial portal vein ligation or sham operation, rats received a 4-day course of either octreotide (15 micrograms/kg in 5% dextrose in water) or placebo (5% dextrose in water) subcutaneously every 8 hours. RESULTS: In portal hypertensive rats, octreotide induced a 13% increase in mean arterial pressure (P < 0.01) and a 19% increase in total peripheral resistance (P < 0.01). Octreotide treatment induced a decrease in extracellular sodium space (22Na injection) (34.2 +/- 0.5 vs. 36.7 +/- 0.4 mL/100 g; P < 0.01) without changes in serum sodium level. In addition, octreotide treatment significantly reduced portal pressure as well as glucagon levels and plasma renin activity. In contrast, octreotide treatment had no effect on mean arterial pressure and extracellular sodium space in shamoperated rats. CONCLUSIONS: Long-term octreotide treatment ameliorated peripheral vasodilatation and sodium retention only in portal hypertensive rats. These findings suggest that in portal hypertension sodium retention can be modified by pharmacological agents that affect peripheral vasodilatation. The specificity of octreotide's effect sheds additional light into the vasodilatory syndrome associated with portal hypertension in liver diseases. PMID- 8425701 TI - T-lymphocyte response to hepatitis C virus in different clinical courses of infection. AB - BACKGROUND: To assess the role played by the immune response in the outcome of hepatitis C virus infection, the CD4+ T-lymphocyte response to viral antigens was studied in infected individuals with different clinical courses. METHODS: Using six recombinant proteins of hepatitis C virus, the study assessed the proliferative responses of peripheral blood mononuclear cells from 41 patients with chronic hepatitis C, 11 patients whose chronic hepatitis was successfully treated with interferon alfa and 11 healthy HCV seropositive individuals. RESULTS: (1) Sixty-five percent of hepatitis C virus-seropositive individuals had CD4+ T-cell responses to viral proteins. (2) All viral proteins were immunogenic for T cells, although NS4 was the most immunogenic. (3) There was a significant correlation between the presence of CD4+ T cell responses to Core and a benign course of infection in healthy seropositives, most of whom were viremic. CONCLUSIONS: CD4+ T-cell responses to Core, although they do not coincide with virus clearance, are associated with a benign course of infection and may be required to maintain humoral and cellular responses protective against the disease. PMID- 8425702 TI - Effects of low-dose captopril on renal hemodynamics and function in patients with cirrhosis of the liver. AB - BACKGROUND: In cirrhotic patients with ascites, captopril has deleterious effects on renal function, which have been referred to as captopril-induced arterial hypotension. The effects of this drug on renal function in cirrhosis were evaluated using low-dose captopril, thereby avoiding any change in arterial pressure. METHODS: In a randomized, double-blind, placebo controlled, cross-over trial, the effects of 12.5 mg captopril on renal plasma flow, glomerular filtration rate (measured by radioisotopic techniques), and sodium excretion in healthy controls and cirrhotic patients with and without ascites were determined. RESULTS: In healthy subjects, captopril only induced a significant, 18% increase in renal plasma flow. In contrast, glomerular filtration rate significantly decreased in patients with (from 108 +/- 7 to 78 +/- 9 mL/min) and without ascites (from 102 +/- 4 to 88 +/- 3 mL/min), whereas renal plasma flow did not change. Urinary sodium excretion also significantly decreased in ascitic patients (from 43.8 +/- 4.4 to 30.6 +/- 3.8 mumol/min). CONCLUSIONS: These data suggest that angiotensin II contributes to maintain renal hemodynamics in cirrhosis with and without ascites. PMID- 8425703 TI - Pathological diagnosis of chronic hepatitis C: a multicenter comparative study with chronic hepatitis B. The Hepatitis Interventional Therapy Group. AB - BACKGROUND: Hepatic histological responses described in hepatitis C virus (HCV) infection include bile duct damage, lymphoid follicles and/or aggregates in portal tracts, large- and small-droplet fat, Mallory body-like material in hepatocytes, liver cell dysplasia and multinucleation, and activation of sinusoidal inflammatory cells. The specificity of these lesions for HCV infection is uncertain. METHODS: In two multicenter trials of recombinant interferon alfa therapy for chronic hepatitis C and B, the frequency of these eight lesions in pretherapy and posttherapy liver biopsy specimens was examined to determine the set of features, if any, that distinguishes HCV from hepatitis B virus (HBV) infection. The lesions were scored in 317 HCV biopsy specimens and 299 HBV specimens. RESULTS: Stepwise logistic regression determined a set of three features more likely to be seen in HCV than in HBV infection: bile duct damage [odds ratio (OR), 4.7; 95% confidence interval (Cl), 1.8-12.3], lymphoid follicles and/or aggregates (OR, 2.4; 95% Cl, 1.2-4.7), and large-droplet fat (OR, 2.4; 95% Cl, 1.4-4.1). A fourth lesion, Mallory body-like material, was seen only in HCV biopsy specimens (OR, 71.6; 95% Cl, 4.4-996.1). CONCLUSIONS: These four histological lesions are useful pathological parameters in the diagnosis of liver disease caused by HCV. PMID- 8425704 TI - Effects of tauroursodeoxycholic acid on cytosolic Ca2+ signals in isolated rat hepatocytes. AB - BACKGROUND: Tauroursodeoxycholic acid (TUDCA) is of potential benefit in cholestatic disorders. However, the effects of TUDCA on cytosolic free calcium [(Ca2+)i], which regulates hepatocyte secretion, are unknown. METHODS: The effect of TUDCA on (Ca2+)i was investigated in groups of isolated rat hepatocytes by microspectrofluorometry and in single cells by confocal line scanning microscopy. RESULTS: Administration of TUDCA (5-50 mumol/L) induced a nearly fourfold increase of basal levels of (Ca2+)i. After a 15 minute treatment period, the TUDCA (10 mumol/L)-induced change in (Ca2+)i was higher than that of other mono-, di-, and trihydroxy bile acids at equimolar concentrations. Pretreatment with TUDCA (10 mumol/L) markedly reduced or abolished increases in (Ca2+)i induced by phenylephrine (1 mumol/L), the microsomal Ca(2+)-translocase inhibitor 2,5-di (tert-butyl)-1,4-benzohydroquinone (25 mumol/L), or taurolithocholic acid (10-25 mumol/L). In Ca(2+)-free medium, TUDCA caused only a reduced and transient increase in (Ca2+)i. TUDCA (10 mumol/L) induced Ca2+ oscillations in all single cells that responded. However, levels of inositol-1,4,5-trisphosphate (IP3) in hepatocytes were not increased by treatment with TUDCA (10 mumol/L). CONCLUSIONS: TUDCA at physiological concentrations potently modulates (Ca2+)i signals in hepatocytes by (1) mobilizing microsomal IP3-sensitive Ca2+ stores by an IP3 independent mechanism, (2) initiating Ca2+ oscillations, and (3) inducing influx of extracellular Ca2+. PMID- 8425705 TI - Chronic pancreatitis: a complication of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE), an autoimmune disease with multisystem involvement, has been reported to be associated with a number of gastrointestinal complications. Acute pancreatitis is an unusual manifestation of this disorder. This report presents two cases of patients with SLE who developed chronic pancreatitis with no other identifiable etiology. PMID- 8425706 TI - Nodular transformation of the liver associated with portal and pulmonary arterial hypertension. AB - A case of multiple focal nodular hyperplasia (FNH) of the liver associated with noncirrhotic portal hypertension and later complicated by pulmonary arterial hypertension leading to death from right heart failure is reported. In retrospect, the portal hypertension diagnosed in early life was most likely due to a congenital hypoplasia of portal vein branches and multiple FNH, a hyperplastic response of the liver parenchyma in association with anomalies of hepatic arterial branches as found within the lesions. This case may represent a form of multiple FNH syndrome restricted to the liver, because neither extrahepatic vascular malformation nor brain tumor was identified at autopsy. The FNH lesions had considerably expanded over the years, and the severe sinusoidal congestion due to chronic right-sided heart failure with subsequent prolonged parenchymal exposure to blood-borne hepatotrophic factors is a likely explanation for both the massive enlargement of FNH lesions and the nodular regenerative hyperplasia observed in the intervening parenchyma. PMID- 8425707 TI - Uptake and transport of macromolecules by the intestine: possible role in clinical disorders (an update). AB - The intestine is exposed to a wide variety of macromolecules. Because macromolecules are antigenic, mechanisms have evolved in the gastrointestinal tract to regulate their absorption. Macromolecular uptake can be beneficial in delivering essential factors for growth and in sampling the antigenic milieu of the gastrointestinal tract. Specific transport mechanisms exist to execute this physiological absorption. However, inappropriate and uncontrolled antigen transport may occur in disease states or as a prelude to disease states in the gastrointestinal tract. Such transport may result in immune responses that are harmful. This review examines physiological transport of macromolecules through epithelia and through M cells. It also considers uncontrolled transport and its relation to disease states. The review concludes with an examination of the interrelationship between antigen transport and an altered immune system in the establishment of gastrointestinal disease. PMID- 8425708 TI - Etiology of fulminant hepatic failure: is another virus involved? PMID- 8425709 TI - Causation of Crohn's disease: the impact of clusters. PMID- 8425710 TI - Will the real cholesterol-nucleating and -antinucleating proteins please stand up? PMID- 8425711 TI - New strategies needed for treatment of primary biliary cirrhosis? PMID- 8425712 TI - Noninvasive sphincter of Oddi testing: what's the score? PMID- 8425713 TI - Ursodeoxycholic acid treatment of intrahepatic cholestasis of pregnancy: observations on efficacy and safety. PMID- 8425714 TI - The silent epidemic of hepatitis C. PMID- 8425715 TI - Aspartate aminotransferase and iron status--lack of support for covariation. PMID- 8425716 TI - Relief of vection-induced motion sickness. PMID- 8425717 TI - Predominance of rectosigmoid neoplasia in ulcerative colitis and its implication on cancer surveillance. PMID- 8425718 TI - Ceruloplasmin inhibits carbonyl formation in endogenous cell proteins. AB - Exposure of cells to oxygen radicals results in cellular injury and protein oxidation. Ceruloplasmin is a plasma antioxidant that increases in concentration during inflammation. Therefore, the ability of ceruloplasmin to protect endothelial cells from neutrophil-mediated injury was investigated. The inhibition of protein oxidation by ceruloplasmin was also examined in neutrophil and endothelial cell proteins by analysis of carbonyl formation. In addition, the iron oxidation state was measured to determine the effect of ceruloplasmin ferroxidase activity in oxygen-radical generating systems. Ceruloplasmin significantly (p < .01) inhibited neutrophil-mediated cytotoxicity of endothelial cells by 48%. Carbonyl formation in phorbol myristate acetate (PMA)-stimulated neutrophil proteins was also significantly (p < .01) reduced by ceruloplasmin from 0.172 +/- 0.028 to 0.086 +/- 0.004 mole carbonyl/mole protein. Even though ceruloplasmin itself had a threefold increase in carbonyl formation (0.452 +/- 0.010 vs. 0.146 +/- 0.018 mole carbonyl/mole protein) in the presence of PMA stimulated compared with unstimulated neutrophils, no loss of functional activity was detected. In xanthine oxidase-treated endothelial cells, ceruloplasmin significantly (p < .05) reduced carbonyl formation from 0.132 +/- 0.010 to 0.097 +/- 0.009 mole carbonyl/mole protein. Ceruloplasmin also significantly (p < .01) oxidized iron when added to PMA-activated neutrophils, thereby decreasing Fe(II) from 98 +/- 8 to 7 +/- 2 microM. Similarly, ceruloplasmin added to xanthine oxidase/hypoxanthine reactions resulted in significant (p < .01) iron oxidation, decreasing Fe(II) from 99 +/- 1 to 15 +/- 3 microM. The ability of ceruloplasmin to protect both endothelial cells and endogenous neutrophil and endothelial cell proteins from oxidative injury suggests that it may be important in regulating cellular and protein damage by oxygen radicals during inflammation. PMID- 8425719 TI - Vasodilatory and toxic effects of spin traps on aerobic cardiac function. AB - The objective of this study was to compare the effect of several structurally related nitrone and nitroso spin traps on the function of the isolated bicarbonate-buffer perfused rat heart model. Spin traps investigated were alpha phenyl-tert-butyl N-nitrone (PBN), alpha-(4-pyridyl-1-oxide)-N-tert-butyl nitrone (POBN), 2-methyl-2-nitroso propane (MNP), 2-hydroxymethyl-2-nitroso propane (MNP/OH), nitrosobenzene (NB), dibromonitrosobenzene-sulfonic acid (DBNBS), and 5,5'-dimethyl-1-pyrroline-N-oxide (DMPO). During perfusion of hearts with increasing concentrations of spin traps, ventricular pressure, coronary flow rate, and heart rate were continuously recorded. The extent of contractile recovery was subsequently measured upon return to spin-trap free perfusion. The percentage of maximum increase in coronary flow with PBN, POBN, MNP, MNP-OH, NB, DBNBS, and DMPO were 11, 40, 45, 66, 28, 28, and 29%, respectively. Thus, all nitroso and nitrone spin traps studied acted as vasodilators. Over the dose range studied, POBN, MNP, MNP/OH, and DMPO did not exert any chronotropic effect. PBN, NB, and DBNBS exerted a negative chronotropic effect at higher concentrations. All spin traps studied, with the exception of DMPO, exerted a negative inotropic effect at the higher concentrations studied. We conclude that all spin traps examined acted as coronary vasodilators. Their negative chronotropic and inotropic effects were minimal in comparison and only manifest at the higher concentrations studied. PMID- 8425720 TI - Effects of physical stress on peroxide scavengers in normal and sickle cell trait erythrocytes. AB - Sickle cell trait subjects are potentially at risk with microvascular complications frequently associated with high altitude and extreme exertion. Studies conducted in our laboratory suggest that exposure of sickle cell trait subjects to treadmill exercise resulted in an increase not only in the susceptibility of red blood cells to oxidation but also in the density of RBC membrane. The trend of increased susceptibility of red cells to oxidation as a consequence of exercise was associated with relatively excessive production of H2O2, inefficient breakdown of H2O2 by GSH-Px- and catalase-catalyzed reactions, and inability/failure to show a transitory increase in the activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH)-generating enzymes. PMID- 8425721 TI - Phosphatidylcholine peroxidation and liver cancer in mice fed a choline-deficient diet with ethionine. AB - A high incidence of hepatocellular carcinoma (HCC) was observed in mice fed a choline-deficient diet containing 0.1% ethionine (CDE) for 19 months. HCC was present in 85% of CDE mice and in 22% of choline-deficient (CD) mice not receiving ethionine. This strong hepatocarcinogenicity of the CDE diet was concomitant with a severe decrease in plasma and liver alpha-tocopherol (Toc) to 60 and 35%, respectively, of those contained in choline-supplemented (CS) control mice. We previously found that this dietary-induced HCC was preceded at 4-week feeding by a depletion of Toc and a remarkable increase of phosphatidylcholine hydroperoxide (PCOOH) in the livers of CDE mice. When HCC was prominent in CDE mice, PCOOH was still elevated. Mouse glutathione S-transferase (GST) M II isozyme, which is related to rat GST-P form, a positive marker for rat hepatic preneoplastic and neoplastic lesions, revealed an inverse histochemical pattern as that seen in rats (i.e., the HCC lesions tended to decreased staining). The aforementioned results taken together indicate that decreases in Toc and enhanced PC peroxidation are important events in CDE-induced mice liver tumors. PMID- 8425722 TI - Distribution of CuZn superoxide dismutase in rat liver. AB - Morphological and cell fractionation approaches were used to establish unambiguously the distribution of CuZn superoxide dismutase (CuZn SOD) in rat hepatocytes. Immunocytochemical observations revealed a primarily cytoplasmic localization of the enzyme. While only trace amounts were found in cell organelles like mitochondria and peroxisomes, lysosomes were labelled stronger than the cytoplasm. The presence of CuZn SOD in lysosomes was also identified in cell fractions of normal and Triton WR-1339-treated rats. Microscopic studies showed that the distribution of CuZn SOD was not affected by Triton, but the Triton treatment induced an apparent increase in the number and size of lysosomes with electron lucent contents which corresponded with a shift of lysosomes to low buoyant density fractions. The majority of CuZn SOD originally present in the mixed mitochondrial-peroxisomal-lysosomal fractions of the normal liver comigrated with the main peak of the lysosomal marker, beta-hexosaminidase, after Triton treatment. The peroxisomal marker, catalase, did not migrate with the CuZn SOD/beta-hexosaminidase-rich fractions in livers from Triton-treated animals. These results confirm earlier observations in rat liver cells, showing that CuZn SOD, a primarily cytosolic enzyme, accumulates in lysosomes. It is not present in significant amounts in rat hepatocyte peroxisomes. PMID- 8425723 TI - Detection of H2O2 release from vascular endothelial cells. AB - Endothelial cells are both significant sources and targets of reactive oxygen species, including O2.-, H2O2, .OH, .NO, and ONOO-, which play important roles in vascular homeostatic mechanisms and pathogenic processes. To better quantify cell oxidant metabolism processes, a fluorescence technique has been developed to measure H2O2 release from bovine aortic endothelial cells. Incubation of H2O2 with horseradish peroxidase (HRP) results in HRP-mediated oxidation of p-hydroxy phenylacetic acid (PHPA) to the fluorescent PHPA dimer, 2,2'-dihydroxy-biphenyl 5,5' diacetate [(PHPA)2]. The HRP-mediated dimerization of 5 mM PHPA with concentrations of H2O2 up to 2.5 mM resulted in a linear increase in fluorescence (R = .995, n = 8). Maximal fluorescence occurred at 2.9 mM H2O2, with greater H2O2 concentrations yielding products with altered spectrophotometric characteristics and decreased fluorescent yield. The fluorescence of (PHPA)2 was pH sensitive and increased 500-fold from pH to 9. Fluorescence versus pH profiles were adjusted to a Henderson-Hasselbalch fitting, with a 50% maximal emission at pH = 8.1 +/- 0.2. The apparent pKa of fluorescence emission correlated well with a weak range of buffering capacity of (PHPA)2, which had a pKa = 8.0 +/- 0.1. With cells maintained in Hank's balanced salt solution (HBSS), the pH can increase to 7.90 during 3 h, with the increased pH due to the loss of HCO3- from HBSS. After adjustment for pH changes, a linear cellular H2O2 release of 217 pmol H2O2.min-1.mg protein-1 was observed. When bovine aortic endothelial cells (BAEC) were incubated with HBSS and PHPA alone, 50% less fluorescence was observed than when HRP was added.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425725 TI - Occurrence of two interdigital neuromas in one foot. AB - In a series of 89 neurectomies, the occurrence of a second primary interdigital neuroma was less than 4%. The clinical presentation and results of surgery were similar to those of a single interdigital neuroma. Two of five patients with a suspected second neuroma in the same foot did not have this entity, but instead had had the incorrect interspace operated on by the previous surgeon. The entity of a second neuroma is truly rare, especially if careful evaluation for correct diagnosis and interspace is done. PMID- 8425724 TI - The modified Brostrom procedure for lateral ankle instability. AB - Twenty-eight ankles in twenty-seven patients (average age 28) underwent the Gould modification of the Brostrom repair for symptomatic lateral ankle instability. Fifty-four percent were high level professional ballet dancers, 35% were recreational athletes, and 11% were nonathletes. Follow-up averaged 64.3 months (range 30-132 months). Of the 28 operations performed, there were 26 excellent results, one good result, and one fair result. All the professional dancers obtained excellent results. There were no failures, stretch-outs, re-dos, or complications. This operation is believed to be an excellent choice for the dancer, athlete, or nonathlete who needs a stable ankle with a full range of plantarflexion and dorsiflexion and normal peroneal function. PMID- 8425726 TI - Evaluation of magnetic resonance imaging in the diagnosis of osteomyelitis in diabetic foot infections. AB - To assess the role of magnetic resonance imaging (MRI) in diagnosing osteomyelitis in diabetic foot infections, 47 diabetic patients with clinical suspicion of osteomyelitis, nonhealing foot ulcer, or soft tissue infection of the foot were examined prospectively by MRI and plain radiographs. Pathological confirmation of diagnosis was obtained in 62 bones from 32 patients. In addition, 14 patients with pathological confirmation of diagnosis underwent technetium-99 MDP triple-phase bone and gallium-67 citrate scans. MRI was significantly more sensitive and accurate (P < .01), with equal specificity in comparison to plain radiographs and technetium and gallium scans. MRI also provided a more detailed and accurate depiction of the anatomy. At early clinical follow-up, complete resection of abnormal bone on an MRI scan correlated with clinical healing. In summary, MRI is indicated when plain radiographs are negative for osteomyelitis or when the extent and accurate depiction of the infective process will facilitate surgical planning. PMID- 8425727 TI - Factors that affect surgical correction in congenital talipes equinovarus. AB - Children with congenital talipes deformity were assessed photographically at birth, and radiologically prior to a standard staged surgical procedure, carried out by one surgeon. A total of 111 feet of 86 children were assessed clinically and radiologically at 4 years of age. Feet with more deformity at birth and poorer preoperative calf muscle required more surgery and had less satisfactory results. Unilaterally affected feet required less surgery and had significantly better postoperative clinical results than bilaterally affected feet. There was a trend toward poorer results in children with a positive family history of talipes, but gender did not affect the outcome. PMID- 8425728 TI - Stress fracture of the proximal phalanx of the great toe. AB - Three cases of stress fracture of the proximal phalanx of the great toe occurring in a 12-year-old female volleyball player, a 17-year-old female long distance runner, and a 12-year-old male soccer player are reported. This condition probably develops after repeated forced dorsiflexion of the first metatarsophalangeal joint with changing steps. When athletes who repeatedly run and jump complain of pain in the first metatarsophalangeal joint, and there is no history of trauma, this condition should also be considered. PMID- 8425729 TI - Bilateral dysplasia epiphysealis hemimelica: a case report. AB - Dysplasia epiphysealis hemimelica is a rare disorder of cartilage growth. Classically, the disease involves a single limb and is isolated to the medial or lateral half of that limb. A patient is described who presented with classic findings of this disease but involvement of both ankles. PMID- 8425730 TI - Proximal Chevron metatarsal osteotomy: single incision technique. PMID- 8425731 TI - Osteochondral lesions of the talar dome. AB - Osteochondral lesions of the talar dome are a common cause of ankle disability. Management options are as numerous as the terms used to describe these lesions. The recognition of a traumatic etiology has increased our understanding and management of these disorders. Modern imaging technology has enhanced the ability to fully evaluate and accurately classify this lesion, which is fundamental for proper treatment. PMID- 8425732 TI - Bunion correction using proximal Chevron osteotomy. AB - Fifty-one cases of moderate to severe bunion deformity with hallux valgus and metatarsus primus varus in 43 patients were treated by bunionectomy, proximal Chevron metatarsal osteotomy, lateral capsulotomy, adductor tenotomy, and lashing of first and second metatarsals together. The hallux valgus angle improved an average of 19 degrees from 33 degrees (mean) preoperatively to 14 degrees (mean) postoperatively. The intermetatarsal angle improved an average of 7.3 degrees from an average of 14 degrees preoperatively to an average of 6 degrees postoperatively. The position of the sesamoids was realigned to beneath the first metatarsal head and the metatarsal length remained essentially unchanged. Union occurred in 9 weeks (mean). No malunions occurred. Foot score profiles revealed a significant improvement in subjective evaluation from 69/100 preoperatively to 83/100 postoperatively with respect to pain, deformity, motion, disability, and cosmesis. Seventy-eight percent of patients had a good to excellent result. Improved subjective evaluations indicated that proximal Chevron osteotomy combined with bunionectomy, capsulotomy, tenotomy, and metatarsal lashing provides a reliable method with respect to stability, technical ease, low complication, and satisfactory surgical outcome for correction of moderate and severe bunion deformity, both as a primary and revision procedure. PMID- 8425733 TI - Technology ... recent survey. PMID- 8425734 TI - Putting patients first. Hospitals work to define patient-centered care. AB - Patient-centered care is quickly becoming a reality across the country, as hospitals begin to define and implement the concept. In a series of articles, we examine the phenomenon and present the results of our exclusive new survey. On page 14, what is patient-centered care, and why has its time come? On page 18, what are the human resources implications for administrators? On page 20, the strategic planning and financial considerations. On page 22, a look at the wide range of facilities implications involved in the patient-centered care concept. PMID- 8425735 TI - Reform follows market. Insurers, providers strengthen ties as debate continues. PMID- 8425736 TI - A new game ahead? Experts ponder prospect of congressional term limits. PMID- 8425737 TI - Standing RBRVS on its head. Educational, operational solutions to MD quandaries. PMID- 8425738 TI - Financial indicators. Hospital expense growth up slightly in 1992. PMID- 8425739 TI - Favorable trend. Low bond rates enable hospitals to save money. PMID- 8425740 TI - Two years and running. The National Practitioner Data Bank begins to roll, but issues remain. PMID- 8425741 TI - Hospitals agree on the need to restructure IS. PMID- 8425742 TI - Data watch. Hospitals offer basic skills education programs. PMID- 8425743 TI - AHA survey: nurse shortage eases dramatically. PMID- 8425744 TI - Let the community decide: a new delivery model. PMID- 8425745 TI - Mondrian and the "boogie woogies": interruption of inner developmental logic or completion in old age? AB - Taking the work of the painter Piet Mondrian as a point of departure, artistic continuity and change are examined from a lifespan developmental perspective. It is argued that decontextualized continuity tends to occur within a given lifestage, whereas contextualized change is apt to emerge during a transition from one lifestage to another. Thus, Mondrian's gradual development of a unique style, predicated on logical, stage-like unfolding, is related to a midlife emphasis on formal structure. From a similar perspective, his dramatic shift away from this inner logic of development in his last work, the Boogie Woogies, is attributed to an ultimate effort in old age to synthesize art and reality, to reconcile a conflict between the laws of art and direct expression of sense experience. PMID- 8425746 TI - A study of autobiographical memories in depressed and nondepressed elderly individuals. AB - An autobiographical memory task was used to study memory processes and depression in elderly individuals. Twenty-seven nondepressed and twenty-seven depressed elderly participants recalled thirty memories. Each memory was self-rated for happiness versus sadness and the degree of importance of the event at the time the event occurred (i.e., "then") and looking back on the event ("now"). Nondepressed participants perceived greater positive change in affective tone between "then" and "now" ratings. Depressed participants recalled more memories rated as sad "now" than nondepressed, and perceived negative and positive memories to become more neutral than nondepressed participants. These results are consistent with a mood congruence hypothesis, in that participants recalled more memories affectively consistent with current mood, and a self-enhancement view of reminiscing, such that recalling memories evaluated as happier was associated with less depression. PMID- 8425747 TI - Predictors of volunteer status in a retirement community. AB - In the present study, actual volunteers, latent volunteers, conditional volunteers, and definite nonvolunteers living in a retirement community are compared on social-structural, role, environmental, resource, lifestyle, and individual difference variables. Three functions emerged from the discriminant analysis. Actual volunteers attend church frequently, are free of activity limitations due to health, have volunteered previously, and belong to several clubs and organizations. Latent volunteers engage in informal religious behaviors, attend church about once a month, are about seventy years old (i.e., they are younger than the other groups), and are very satisfied with their neighborhood. Conditional volunteers engage infrequently informal religious behavior, have no instrumental activity limitations due to health, did not attend college, have very high neighborhood satisfaction, and are seventy-four years old (i.e., older than latent volunteers and actual volunteers). Implications for recruiting latent and conditional volunteers are discussed. PMID- 8425748 TI - The association of positive and negative events with depressive symptomatology among caregivers. AB - A pilot study of eighty-two caregivers was conducted in South Carolina in 1991 to identify positive and negative factors associated with caregiving. Through home visits, interviewers obtained data on a variety of physical and mental health measures, including two new scales designed to measure perceived "positive" and "negative" events that had occurred in the previous month. The Center for Epidemiologic Studies Depression scale was used as a measure of depressive symptomatology. For the new scales, only items that were significantly correlated with depressive symptomatology (p < 0.01) were retained. The new "positive" event scale (8 items) and the new "negative" event scale (16 items) had alpha coefficients of 0.79 and 0.86, respectively. These scales may be useful to researchers in sorting out mediating factors related to the burden of caregiving and in providing points for intervention. PMID- 8425749 TI - Neighborhood deterioration and social isolation in later life. AB - The purpose of this study is to test a conceptual model that attempts to show how selected neighborhood characteristics are related to social isolation in later life. This model specifies that older adults with low levels of educational attainment are more likely to experience financial problems and that elderly people who are confronted by financial difficulties are more likely to reside in dilapidated neighborhoods. The model further predicts that deteriorated neighborhoods in turn tend to promote distrust of others and older adults who are more distrustful of others tend to be more socially isolated. Based on data provided by a nationwide survey of elderly people, subsequent analyses revealed strong support for the theoretical sequence described above. PMID- 8425750 TI - A model for metastases formation. AB - Our aim is to propose a mathematical model for the forming of metastases resistant or not, from their birth, to one drug or a combination of drugs, associated with a new spontaneous tumorous growth model. Practical conclusions are then drawn relating to the effectiveness of chemotherapy. PMID- 8425751 TI - A GAUSS program for computing the Foulkes-Davis tracking index for polynomial growth curves. AB - We have previously published a GAUSS program for computing the Foulkes-Davis tracking index, gamma, from a one-sample longitudinal data set when no assumptions were made concerning the structure of the individual growth curves (Schneiderman et al., Am J Hum Biol, 4 (1992) 417-420). In this paper we consider the computation of the Foulkes-Davis index assuming that each individual growth curve may be adequately represented by a polynomial function in time and a GAUSS program performing these computations is made available. As with the two other tracking indices we have described, gamma and kappa (Schneiderman et al., Am J Hum Biol, 2 (1990) 475-490), this one can be used to evaluate regularity in patterns of growth or adaptation. An example is presented where statural growth in the same three groups considered in the earlier papers are analyzed. The small disparities between these and the earlier results are discussed in view of the different assumptions of the models and the differences in how they operationalize the concept of tracking. PMID- 8425752 TI - A functional model of the mammalian circadian pacemaker. AB - This paper describes a general model of the mammalian circadian pacemaker with specific parameters for golden hamsters. The model successfully simulates free runs in constant darkness and constant light, phase shifts induced by light pulses and entrainment and relative coordination under light-dark cycles. It also accounts for the recent findings that the amplitude of the phase response curve to light pulses is dependent on the free-running period of the animal and on the previous history of photic stimulation. Effects of non-photic stimuli on the circadian pacemaker are discussed. PMID- 8425753 TI - A model for blood flow through a stenotic tube. AB - This article deals with the introduction of the modified Casson's fluid model as the true representation for the blood for the steady laminar flow through a small diameter artery with axi-symmetric identical double stenoses in series. The governing equations are solved by using the finite element method. The results for the velocity profiles, the pressure and the wall shear stress distributions in addition to the location and length of the flow reversal zones have been brought out and discussed in reference to the severity of the disease. It has been observed that the non-Newtonian nature of the blood helps in reducing the magnitude of the peak wall shear stress at the throat and the length of the reversed flow regions in the post stenotic dilatation. PMID- 8425754 TI - Animated visualization of a high resolution color three dimensional digital computer model of the whole human head. AB - The interactive visualization of animated images through a computerized three dimensional (3D) full color model of an unstained cadaveric human head is presented. Serial full color images were taken of the blockface of a cryomicrotomed frozen human head every 200 microns. From this series of images a three dimensional digital model with a resultant pixel resolution of 200 microns3 was created on a UNIX workstation. Using this database, resampled images were computed along orthogonal axes and written sequentially to a write-once-read-many times (WORM) videodisc unit. Playback of this customized videodisc dataset provides animations of the digitally reconstructed slices and 3D reconstructed surface models. An interactive interface to the animated sequences is provided through a PC based tutorial package. This tutorial program is able to access videodisc frames to display animations and labeled still images in a software window to illustrate various neuroanatomic topics. The technique of animation as applied to this high resolution 3D model provides insight into complex spatial relationships and has great potential in research and as a teaching tool in the neurosciences. PMID- 8425755 TI - Incidence pattern of thyroid cancer in Norway: influence of birth cohort and time period. AB - The incidence of thyroid cancer in Norway increased about 2-fold for both sexes during the period 1955 to 1989 (4691 cases). In the last 5-year period, however, a decline was observed, especially among females. The ratio between age-adjusted incidence rates in Northern Norway compared with Southern Norway was 1.6 for females and 1.5 for males, and the incidence pattern during the period was similar in the 2 regions. The analysis of age-cohort-period models showed a strong cohort effect in both sexes. The reason for this is not clear, although radiation treatment during childhood and dietary habits may possibly be involved. A weaker but statistically significant and transient period effect was also present, giving relatively higher incidence rates, especially in the 1970s. Although this result may be consistent with an influence of radioactive fallout in the northern area, more detailed studies are needed. PMID- 8425756 TI - Linkage mapping in familial breast cancer: improved localisation of a susceptibility locus on chromosome 17q12-21. AB - Fifteen pedigrees with a total of 75 cases of breast cancer, 10 of ovarian cancer and 53 of other cancers have been collected. Polymorphic markers on chromosome 17q have been screened to locate a putative breast-cancer gene using DNA from relevant individuals within these families. Pairwise LOD scores have been calculated for markers CMM86, NM23, 42D6 and MFD188. The maximal summated LOD for the 15 families is 4.45 at theta = 0.025 using 42D6. All cases of bilateral breast cancer and ovarian cancer appear to be linked to this region. Recalculating LOD scores on the assumption of linkage in these cases increases the maximal summated LOD to 5.62 at theta = 0.025 using 42D6. A genetic exclusion map of critical recombinants in linked families suggests that the gene is flanked by markers 42D6 and MFD188, a region 5 to 10 cm in length. PMID- 8425757 TI - Long-term impact of reproductive factors on cancer risk. AB - The relationship between reproductive variables (parity, age at first birth, number of induced and spontaneous abortions) and cancer risk has been analysed using data from an integrated series of case-control studies conducted in northern Italy between 1983 and 1992. The overall data-set included women below age 75 with histologically confirmed cancers of the following sites: oesophagus, 58; stomach, 280; colon, 405; rectum, 210; liver, 82; gall-bladder, 29; pancreas, 129; breast, 3,415; cervix, 742; endometrium, 725; ovary, 953; bladder, 68; kidney, 56; thyroid, 180; lymphomas, 80; myelomas, 57; and a total of 5,619 controls admitted to hospital for acute non-neoplastic, non-gynaecological, non hormone-related conditions. Multivariate odds ratios, as estimators of relative risks (RR), were obtained after allowance for age, education, use of oral contraceptives and oestrogen replacement treatments, plus various reproductive factors. Direct significant trends with parity were observed for cancer of the liver (RR for women with > or = 4 births vs. nulliparae = 3.3) and cervix uteri (RR = 4.1). The risk of gall-bladder cancer was also elevated for multiparae (RR = 1.9). No significant inverse trend in risk emerged. However, the RRs in multiparae were significantly below unity for breast (RR = 0.8), endometrium (RR = 0.7), and ovary (RR = 0.8). With reference to age at first birth, a significant trend in risk was observed for breast cancer (RR = 1.4 for 25 to 29 and 1.5 for > or = 30 vs. < 25 years). In contrast, the risk of cervical cancer was inversely related to age at first birth. For spontaneous abortions, the only significant inverse trend was for ovarian cancer (RR = 0.7 for > or = 2 vs. 0 abortions), but also the point estimate for endometrial cancer in women with > or = 2 abortions was below unity. For induced abortions, there was a strong inverse trend in risk for endometrial cancer (RR = 0.5), and the RRs were below unity also for colon and breast cancer. In contrast, cervical cancer was directly associated with the number of spontaneous abortions. Although the underlying aetiological interpretations are different for various cancer sites, this study provides, in a large and uniform data-set, quantitative information on the long-term impact of reproductive factors on cancer risk. PMID- 8425758 TI - Evaluation of the new (1987) TNM classification for thoracic esophageal tumors. AB - A total of 351 patients with thoracic esophageal carcinoma were prospectively classified according to the new (1987) TNM classification. Sixty-two patients received chemotherapy and/or radiotherapy without surgery. Esophagectomy was performed on 291 patients, among whom 139 underwent cervical, mediastinal and abdominal lymph adenectomy (3-field dissection). The number of stage IIB patients was unnaturally small, and the 3-year survival rates of stages IIA and IIB were similar. The survival curves for patients of T, N and M categories distributed well except for those of stages IIA and IIB. Numbers of patients in N and pN categories and those in M and pM categories showed poor coincidence. The 5-year survival rate for MI patients (10.5%) was too good, which suggested the mingling of patients with rather better prognosis. When MI (LYM) patients were excluded from MI, the 3-year survival rate fell to 0.3%. The 5-year survival rate for pMI (LYM) patients who underwent 3-field dissection was 38.2%. Survival rates and numbers of positive nodes showed negative correlation in patients who underwent 3 field dissection. According to these results, we propose the following revision in the next TNM classification: (1) to group stage IIA and stage IIB together to form Stage II; (2) to include cervical and coeliac lymph nodes among the regional lymph nodes, or to designate metastasis in non-regional lymph nodes separately from metastasis in viscera; and (3) to divide N1 into N1 and N2 according to the number of positive lymph nodes. PMID- 8425759 TI - Sinonasal cancer and occupational exposure to formaldehyde and other substances. AB - A case-control study of cancer of the nose and paranasal sinuses was conducted in France to determine whether occupational exposure to formaldehyde was associated with an increased risk of sinonasal cancer. Exposures to 14 other substances or groups of substances were also studied (wood dust, leather dust, textile dust, flour dust, sugar dust, coal/coke dust, nickel compounds, chromium compounds, chromium VI, welding fumes, soldering fumes, cutting oils, paints and lacquers, glues and adhesives). Cases (n = 207) and controls (n = 409) were interviewed to obtain detailed information on job history and other potential risk factors for sinonasal cancer. In addition, a questionnaire specially designed for this study was used to help assess exposures to formaldehyde and other substances of interest. The questionnaires were translated into history of occupational exposure by an expert in industrial hygiene, without knowledge of case-control status. Several exposure variables (lifetime average level, duration, cumulative level) were used to describe the risk related to exposure to formaldehyde. Potential confounding factors (occupational and non-occupational) were examined and adjusted for when necessary. No significant association was found between exposure to formaldehyde and squamous-cell carcinomas of the sinonasal cavities. Because of the strong association between exposure to wood dust and nasal adenocarcinoma, it was not possible to assess an independent effect of formaldehyde on this type of cancer. However, among males exposed to medium or high levels of wood dust, the risk of adenocarcinoma associated with formaldehyde was significantly elevated for the highest exposure categories for average level (OR = 5.3, 95% confidence interval = 1.3-22.2), cumulative level (OR = 6.9, 95% CI = 1.7-28.2) and duration of exposure (OR = 6.9, 95% CI = 1.7-27.8). Although a residual confounding effect of wood dust could not be excluded, this study suggests that exposure to both formaldehyde and wood dust may increase the risk of nasal adenocarcinoma, by comparison with the risk due to wood dust alone. This study also indicated an increased risk among males who had been exposed to glues and adhesives, for all histologic types, which was not explained by a confounding effect of paints and lacquers, wood dust or formaldehyde. No other significant association was observed. PMID- 8425760 TI - Site distribution of cutaneous melanoma in Queensland. AB - The age-adjusted incidence rates of cutaneous melanoma in Queensland in 1987 have been analysed for 16 anatomic sites, taking into account their surface areas. In men, the incidence of invasive melanoma on the ears, a chronically sun-exposed site, was extraordinarily high with annual rates of over 200 per 10(5) units of surface area in the Queensland population. Next highest rates of over 100 melanomas per 10(5) units were found on the face, neck, shoulders and back in men and the face and shoulders in women. Comparison with site-specific incidence rates in the same population 7 1/2 years previously showed that incidence of invasive disease had significantly increased for all these sites, though the largest relative increase in this period occurred on the forearm in both men and women. Melanoma was very rare on the buttocks of both sexes and on the scalp in women, sites which receive the least sun exposure. These findings are consistent with the theory that excessive total sun exposure plays a major role in the aetiology of cutaneous melanoma. PMID- 8425761 TI - Different roles for phenacetin and paracetamol in cancer of the kidney and renal pelvis. AB - A population-based case-control study of kidney cancer was carried out in New South Wales using data from structured interviews with 489 cases of renal-cell cancer and 147 cases of renal pelvic cancer diagnosed in 1989 and 1990, together with 523 controls from the electoral rolls. This study showed that the risk of renal pelvic cancer was increased by phenacetin/aspirin compound analgesics (RR = 12.2; 95% CI 6.8-22.2) to a far greater extent than by paracetamol (RR = 1.3; 95% CI 0.7-2.4; not significant). There was a doubling of risk (RR = 2.0; 95% CI 0.9 4.4) in the highest tertile of paracetamol taken in any form compared with values for non-users of any type of analgesic. By contrast, the risk of renal-cell cancer appeared to be increased to a similar degree by phenacetin/aspirin compound analgesics (RR = 1.4; 95% CI 0.9-2.3) and paracetamol taken in any form (RR = 1.5; 95% CI 1.0-2.3). When both drugs were treated as alternative forms of the same risk factor, the risk was increased by 1.7 (95% CI 1.2-2.4). On this evidence, we postulate that phenacetin/aspirin compounds are weakly carcinogenic in the renal parenchyma through the metabolic conversion of phenacetin to paracetamol, and potently carcinogenic in the renal pelvis by different or additional pathways involving renal papillary necrosis. In addition, there is an indication of a weak link between paracetamol and renal pelvic cancer. PMID- 8425762 TI - K-ras mutations in human adenocarcinoma of the lung: association with smoking and occupational exposure to asbestos. AB - We investigated point mutational activation of the ras genes (K-ras codons 12, 13 and 61; N-ras codons 12, 13 and 61; H-ras codons 12 and 61) in primary, resected lung cancer by dot blotting and oligonucleotide hybridization. K-ras mutations were found in 14 (29%) of the 48 lung tumour specimens examined, but no N-ras or H-ras mutations were found. The highest frequency of K-ras mutation was observed in adenocarcinoma: 12 of the 21 samples studied (57%) had a mutation, which is one of the highest frequencies reported for lung adenocarcinoma. The commonest type of mutation in these lung tumour samples consisted of transversions: we observed 11, of which 8 (57% of all mutations) were G to T transversions. Most of the 48 patients studied had a history of heavy smoking, either with or without evidence of occupational exposure to asbestos. Statistical analysis revealed--in addition to the highly significant association between the adenocarcinoma type of lung cancer and K-ras mutation--a clear association of K-ras mutations with heavy life-time smoking (> or = 50 pack-years of cigarette smoking; odds ratio (OR) 4.9, 90% CI 1.2-19.5, multivariate analysis). In addition, occupational asbestos exposure showed an elevated, but non-significant, OR of 2.2 (90% CI 0.6-8.7) with the presence of K-ras mutation. We conclude that the occurrence of K-ras mutations in adenocarcinoma of the lung is frequent, and that such mutations are associated with heavy life-time exposure to tobacco smoke, possibly in combination with occupational exposure to asbestos fibres. PMID- 8425763 TI - Prothymosin alpha restores depressed allogeneic cell-mediated lympholysis and natural-killer-cell activity in patients with cancer. AB - Cancer-bearing patients exhibit a variety of profound T-cell abnormalities which include decreased cytotoxic capacity as measured by allogeneic cell-mediated lympholysis (CML), natural-killer (NK) cell activity, and decreased lymphokine production. In patients with advanced solid malignancies, allogeneic CML, tested by a 4-hr 51Cr-release assay, was significantly lower than in a group of normal individuals. If optimal doses of affinity-purified prothymosin alpha (ProT alpha) were present during mixed lymphocyte culture, the CML of cancer patients was increased almost to normal levels. Mixed lymphocyte reaction, tested by tritiated thymidine uptake, was also decreased in these patients and was enhanced to normal levels if ProT alpha was added to the cultures. NK activity was decreased in these patients according to 51Cr-release assays. ProT alpha increased the NK activity up to normal levels. The reduced NK and CML activities in cancer patients were associated with abnormal production of prostaglandin E2 (high) and interleukin-2 (low), which were to a great extent normalized in the presence of ProT alpha. These results demonstrate that ProT alpha is capable of potentiating or fully restoring the deficient cytotoxic effector function of peripheral mononuclear cells (MNC) in patients with advanced malignancies. PMID- 8425764 TI - Myoepithelial and basement membrane antigens in benign and malignant human breast tumors. AB - Serial cryostat sections of 160 human breast lesions and of 9 lymph-node metastases were studied by indirect immunofluorescence. We used monoclonal antibodies (MAbs) to lining-epithelium-specific keratin 8 and to myoepithelium specific keratin 17 in combination with polyclonal and monoclonal antibodies to major basement membrane components, laminin, collagen type IV, entactin/nidogen, and large heparan sulfate proteoglycan (perlecan) core protein. Continuous basement membranes adjacent to a basal layer of keratin-17-positive myoepithelial cells were typical for normal, benign and in situ carcinomatous structures. In invasive and metastatic structures, always formed by keratin-8-positive tumor cells, basement membranes were found only rarely and with conspicuous fragmentations. This lack of basement membranes correlated with loss of myoepithelium identified by staining for keratin 17. In comedo structures of invasive ductal carcinomas and in papillary carcinomas, fibrovascular complexes with numerous blood vessels and deposition of basement membrane material were often seen in the stroma. Immunomorphological analysis of 41 cases of doubtful diagnosis at intra-operative biopsy was also performed. A combination of MAbs to keratins 8 and 17, and to basement membrane components, made it possible to distinguish between morphologically similar benign and malignant proliferations and to detect single-cell invasion of the stroma. This combination of antibodies may be recommended as an auxiliary immunomorphological tool for differential diagnosis of intra-operative breast biopsies in dubious cases. PMID- 8425765 TI - Thymosin beta-10 expression in melanoma cell lines and melanocytic lesions: a new progression marker for human cutaneous melanoma. AB - When screening a subtraction library for sequences that were specifically expressed in highly metastatic human melanoma cell lines, a cDNA clone was isolated encoding thymosin beta-10. We found that expression of thymosin beta-10 mRNA was associated with metastatic behavior of various human melanoma cell lines in nude mice. Furthermore, Northern blot analysis showed that also in freshly harvested human melanocytic lesions thymosin beta-10 was differentially expressed. Although expression of thymosin beta-10 was also examined in other non melanoma model systems and materials, no clear relation could be established with metastatic potential or malignancy. Therefore, we conclude that thymosin beta-10 can be considered as a new progression marker for human cutaneous melanoma. PMID- 8425766 TI - A monoclonal anti-idiotypic antibody with an internal image of a "phosphatidylinositol-like" structure derived from anti-"phosphatidylinositol like" IgG from the sera of patients with proliferative malignancies. AB - Using an enzyme-linked immunosorbent assay (ELISA) with phosphatidylinositol (PtdIns) coated on well-plates, high levels of anti-"PtdIns-like" autoantibodies (autoAbs) have been previously described in sera of cancer patients. These anti "PtdIns-like" autoAbs were purified and injected into BALB/c mice. A monoclonal anti-idiotypic antibody (Ab2), internal image of human endogenous "PtdIns-like" structure called AIPI, was selected. Then the immunological binding in sera of cancer patients was evaluated on AIPI coated on well-plates. Using this indirect ELISA method, we found a statistically highly significant immunological binding in sera of patients with epithelial tumors, which correlated with that previously found with the PtdIns molecule coated on well-plates. Moreover, AIPI mimics an endogenous structure closely associated with PtdIns specifically encountered in epithelial proliferative disease. PMID- 8425767 TI - Inhibition of growth-factor-activated proliferation by anti-estrogens and effects on early gene expression of MCF-7 cells. AB - Recently, it was reported that the anti-estrogen tamoxifen not only inhibits estradiol-stimulated growth of MCF-7 cells but also significantly reduces the proliferation rate of cells stimulated by growth factors. We have confirmed this finding and also shown that the new anti-estrogen droloxifene inhibits the proliferation of epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I)-stimulated MCF-7 cells. The growth-factor-induced proliferation was inhibited in a dose-dependent manner by the anti-estrogens in the complete absence of estrogen and FCS. Of the anti-estrogens, droloxifene was considerably more potent than tamoxifen. Because the expression of the proto-oncogenes c-fos and c-myc has been considered a key event in development of the mitogenic response, we examined the effects of anti-estrogens on c-myc and c-fos gene expression. We included in these investigations the steroidal anti-estrogen ICI 164,384 because this compound has no or very little estrogenic activity. The studies revealed that all 3 anti-estrogens transiently induced c-myc mRNA expression. However, the anti-estrogens inhibited estradiol-induced c-myc mRNA expression, although with different potencies. Pre-incubation of MCF-7 cells with droloxifene and tamoxifen resulted in elevated levels of growth-factor-induced c myc mRNA expression. In contrast, the anti-estrogens did not induce c-fos mRNA or affect the expression of c-fos mRNA induced by growth factors. In conclusion, non steroidal anti-estrogens inhibit growth-factor-stimulated proliferation of MCF-7 cells without inhibiting growth-factor-induced c-myc or c-fos mRNA expression. PMID- 8425768 TI - Cancer-cell traffic in the liver. II. Arrest, transit and death of B16F10 and M5076 cells in the sinusoids. AB - Fluorescent probes were used to detect BUdR-labelled B16F10 and M5076 cancer cells delivered to the livers of mice via intrasplenic injection. In liver sections stained for succinic dehydrogenase, which permits the periportal, acinar zone 1 to be distinguished from the pericentral zone 3, counts were made of the zonal distribution of fluorescent, intact cancer cells and, by default, the numbers of "lost" cells. Very few intact cancer cells leave the liver from the single bolus of the intrasplenic injection, and even fewer of these generate pulmonary lesions; therefore, within the time limits of these experiments, the liver is virtually a closed system. A dynamic view of intrahepatic cancer-cell traffic with respect to zones 1 (periportal) and 3 (pericentral) was obtained from static measurements of cell densities at different times after intrasplenic injection, by means of Markov chain probability analysis. This indicated that, during the first hour after arrival in zone 1 of the liver sinusoids, there is a 10% probability of a B16F10 cell remaining intact in zone 1, an 89% probability of cell death in zone 1 and only a 1% probability of the cell passing into zone 3. During the same period, there is a 77% probability of an M5076 cell remaining intact in zone 1, a 21% probability of death, and a 2% probability of relocation to zone 3. In both cell types, very few cells were lost from zone 3. Further proportional death in zone 1 diminished over the next 23 hr, concomitant with an increased proportion of cell death in zone 3. Our results indicate that, although there is considerable variation between the 2 cell types studied here, most (B16) or many (M5076) of these cancer cells entering the liver via the portal vein die within 1 hr in zone 1 of liver lobules. In addition, very few of the cells delivered to zone 1 travel along the sinusoids to zone 3, and few of these reach the lungs in a viable state. PMID- 8425769 TI - Increased radioresistance of EJras-transformed human osteosarcoma cells and its modulation by lovastatin, an inhibitor of p21ras isoprenylation. AB - Alterations in ras oncogene expression have been associated with increased cellular resistance to ionizing radiation. As an extension of studies with murine cell models, we have now explored the radioresponses of human osteosarcoma (HOS) sub-clones that differ in their EJras expression. Quantitative analysis revealed a tight correlation between the amounts of ras-encoded mRNA and p21 produced, and the degree of cell radioresistance. Interestingly, treatment of the ras transformed cells with lovastatin, an inhibitor of p21ras post-translational processing via the mevalonate pathway, markedly decreased their radioresistance. Under the experimental conditions used, lovastatin prevented the membrane association, but not the biosynthesis, of p21. The decline in radiation resistance following lovastatin treatment could not be attributed to perturbation of cholesterol metabolism or to non-specific cell-cycle effects. In agreement, lovastatin did not alter the radiation responses of control HOS cells that do not express EJras, or those with an activated met oncogene. The results indicate that elevation in ras gene expression can lead to increased radioresistance of human tumor cells. It appears, however, that p21ras membrane localization is critical for maintenance of the radioresistant phenotype, thus providing a target for pharmacological intervention. PMID- 8425770 TI - Establishment of L1210 leukemia cells resistant to the distamycin-A derivative (FCE 24517): characterization and cross-resistance studies. AB - N-deformyl-N-[4-N,N-bis(2-chloroethylamino)benzoyl] distamycin-A (FCE 24517) is a new cytotoxic anti-tumor agent in phase-1 clinical trials. We have isolated stable FCE-24517-resistant cell sublines from murine leukemia L1210 cells by in vitro exposure to the drug. FCE 24517 selects a mixed population of resistant cells: the L1210/24517(1) cell line in vitro was in fact resistant to the selecting agent (RI 48.3), as well as to L-PAM (RI 5.4) and DX (RI 8.6) and over expressed the mdr-I gene. When L1210/24517(1) cells were implanted in vivo and evaluated for sensitivity to the same agents, resistance was observed only to FCE 24517 and partially to L-PAM, whereas DX had the same anti-tumor efficacy as on the sensitive line. The clone derived from the above subline (L1210/24517(2)) was resistant to FCE 24517, distamycin-A and other cytotoxic compounds bearing the distamycin-A skeleton, and fully sensitive to DX and other anti-tumor compounds involved in the multi-drug resistance mechanisms, with a complete disappearance of the mdr phenotype. L1210/24517(2) cell line is partially cross-resistant to L PAM, this resistance being accounted for by higher GSH intracellular levels, which however do not influence the resistance to FCE 24517. In fact, BSO treatment was capable of significantly modifying only the cytotoxicity of L-PAM. Our data suggest that L1210/24517(2) cells present a mechanism of resistance specific for FCE 24517 and related molecules. PMID- 8425771 TI - Expression of functionally intact PDGF-alpha receptors in highly metastatic 3LL Lewis lung carcinoma cells. AB - The ability of disseminating tumor cells to grow in a target organ is the final limiting step of the metastatic cascade. The growth of a highly lung-metastatic clone, D122, of the murine 3LL Lewis lung carcinoma was induced in vitro with lung-conditioned media (CM) to a greater extent than that of a weakly metastatic clone, A9. With the use of platelet-derived growth factor (PDGF)-alpha-receptor specific antibodies, the possible paracrine mode of metastatic cell growth was further suggested by demonstrating the presence of these receptors on highly metastatic cells only. Receptors for PDGF have been found almost exclusively on cells of mesenchymal and glial origin. Therefore, the functionality of this receptor in the D122 epithelial cell line was verified by immune complex kinase and ligand-stimulation assays. Moreover, PDGF was shown to specifically induce in vitro growth of D122 highly metastatic cells only and to be abundantly expressed in lung CM but not in kidney or liver CM. Thus, the interaction of PDGF-like factors in the lung with PDGF receptors on the metastatic tumor cell may be important in the development of metastatic lesions in the target organ. PMID- 8425772 TI - Hypersensitivity of human testis-tumour cell lines to chemotherapeutic drugs. AB - Metastatic testis tumours, in contrast to most other types of cancer, can be cured by drugs. To investigate which classes of chemotherapeutic drug are differentially toxic to testis-tumour cells, we compared the in vitro dose response curves of 5 human testis and 5 bladder-cancer cell lines to 12 compounds. The testis cells were hypersensitive to drugs that interact directly with DNA (m-amsa, bleomycin, cisplatin, doxorubicin, methylnitrosourea, mitozolomide, etoposide, mitomycin-C), but little or no difference between the 2 cell types was seen following exposure to drugs whose mechanisms of action do not involve direct interaction with DNA (methotrexate, 5-fluorouracil, colchicine, vinblastine). We conclude that testis tumour cells are either less tolerant of, or have a reduced capacity to repair, DNA damage. PMID- 8425773 TI - Multiple personality disorder and iatrogenesis: the cautionary tale of Anna O. AB - An examination of Breuer's treatment of Anna O. illustrates some of the controversies surrounding the recent rise of case reports of multiple personality disorder. Anna O., the first patient of the cathartic method, psychoanalysis, and dynamic psychiatry, fits current criteria for multiple personality disorder. Breuer's treatment, however, may have contributed to her states of absence; the timing, type, and intensity of Breuer's interventions make it possible that he unwittingly encouraged and amplified Anna's dissociations, reified her ego fragments, and then explained Anna's symptoms with the pseudomemories and confabulations recovered from Anna while she was hypnotized. A review of Breuer's treatment highlights some of the controversial aspects of multiple personality disorder, specifically its possible vulnerability to iatrogenesis via suggestion and unconscious collusion and other factors. The current stance of some multiple personality disorder enthusiasts, opaque to their participation in interactions that may lead to certain patient productions, resembles the older psychoanalytic stance exemplified by the early Breuer and Freud. The dialectic of the therapist as a neutral observer versus as an influential participant continues to be a focus of controversy, both within psychoanalysis and psychotherapy and in the understandings of the etiology and treatment of multiple personality disorder. PMID- 8425774 TI - The Waterloo-Stanford Group C (WSGC) scale of hypnotic susceptibility: normative and comparative data. AB - The Waterloo-Stanford Group C (WSGC) hypnotic susceptibility scale was developed as a substitute for the individually administered Stanford Hypnotic Susceptibility Scale, Form C (SHSS:C). A first investigation with WSGC reports normative data on 259 subjects, and the results indicate that it is comparable in most important respects to the norms of SHSS:C. A second investigation directly compared WSGC and SHSS:C in a counterbalanced design on 65 subjects, and the two scales correlated .85. It is argued that, when used as a follow-up to the Harvard Group Scale of Hypnotic Susceptibility, Form A, WSGC provides a valid criterion of hypnotic ability. PMID- 8425775 TI - The hypnosis scales at their centenary: some fundamental issues still unresolved. AB - Current approaches to the measurement of hypnotic performance can be traced back to the 19th century. In part because of these early origins and in part because of the nature of hypnotic phenomena, the hypnosis scales are unique psychometric instruments. The classic hypnosis scales are based on the notion of a "performance ladder"; items are scored on a pass/fail basis and can be arranged in increasing order of difficulty. Some of the implications on this "performance ladder" approach are reviewed. The evidence for two-mechanism models of hypnotic performance is reviewed. It is argued that this kind of formulation is at least as plausible as one that argues that the hypnosis scales measure "one thing" or "mostly one thing." If it were the case that the hypnosis scales were tapping two different and distinct processes, the label "hypnotic susceptibility" could not be unambiguously applied to scores on the hypnosis scales. The hypnosis scales would appear well-suited to the investigation of underlying mechanisms, yet no consistent picture of the mechanisms underlying hypnotic performance on the scales has emerged thus far. No resolution is presented, but some of the reasons why such a resolution is so elusive are discussed. The future of hypnosis scales is discussed with respect to multidimensional assessment and alternatives to the "work sample" approach. PMID- 8425776 TI - The Carleton Skills Training Package for modifying hypnotic susceptibility--a replication and extension: a brief communication. AB - This study employed the Carleton Skills Training Package (CSTP) to attempt to enhance both objective and subjective components of hypnotic susceptibility. In addition, changes in susceptibility were compared for subjects administered a standard hypnotic induction procedure and for subjects given brief "place yourself in hypnosis" instructions. Results indicated that subjects who were administered the CSTP exhibited significant gains in both objective and subjective susceptibility scores that were maintained at two separate posttests with different scales. No differences were observed between the groups administered the standard induction and those administered the self-induction instructions. PMID- 8425777 TI - Modeling the behavior and attributes of injecting drug users: a new approach to identifying HIV risk practices. AB - The behaviors and attributes of 503 Scottish injecting drug users were modeled using the linear structural equations program LISREL. Drug use was directly related to prison experience, sexual activity, sharing of injecting equipment, and prostitution. Although the prevalence of HIV among the sample was low (2.0%), the pattern of risk behaviors observed in the data affords potential for future spread of the virus. Harm reduction measures taken by injectors in response to the threat posed by AIDS were inversely related to drug use but, more encouragingly, directly related to awareness of the disease, treatment for drug use, and prostitution. PMID- 8425778 TI - Using scripted cooperation to communicate information about the consequences of alcohol and cocaine use. AB - Fifty-two participants from undergraduate psychology classes at a private university were asked to study, either cooperatively or individually, information dealing with the consequences of using alcohol and cocaine. Objectives were to (a) determine the effect of cooperative study on recall, (b) assess personal reactions to the information presented, and (c) assess the effect of individual differences. Findings indicate that cooperative study can facilitate both recall of information as well as degree of understanding and confidence in dealing with persons using alcohol/cocaine; that the materials used were received positively; and that verbal ability may play a subtle role in college students' responses to alcohol/cocaine information. PMID- 8425779 TI - The effect of living abroad on alcohol expectancies among American adolescents in Germany. AB - Previous research has indicated that living abroad has a mitigating effect on alcohol use among American adolescents. Self-reported reasons for drinking and alcohol expectancies of American high school students who have lived abroad for 2 years or less were compared to those of American high school students who have lived abroad for over 10 years. Results indicated that students who have lived abroad for over 10 years endorse social and pleasure seeking and tension reduction reasons for drinking less often than students who have spent 2 years or less outside of the United States. Implications for preventive programs are discussed. PMID- 8425780 TI - A mathematical model of habituation and addiction. AB - A model describing changes in the frequency of drug use for an individual is presented. It is shown that the model predicts the evolution of a large number of long-time users who will exhibit behavior similar to heat flow, with the percentage of users per unit frequency analogous to temperature. Data from the NIDA 1985 Household Survey which suggest such behavior among users are presented, and they are shown to be consistent with a principle of increasing entropy. PMID- 8425781 TI - Health services research: what is it? AB - This article reviews the hegemonic understanding of health services research in the United States and in the Anglo-Saxon world, taking as a point of reference the recently published PAHO anthology, Health Services Research: An Anthology (which contains the classics in health services research). The author outlines the main characteristics of health services research in the United States, and criticizes the focus on medical care of most health services research literature and the narrow spectrum of positions presented in that literature. He concludes that there is an urgent need for analyses of the socioeconomic and political forces that determine the level of health and the type of health services that exist in any society, analyses that are usually avoided in health services research. PMID- 8425782 TI - Sugar and spice and everything nice: health effects of the sexual division of labor among train cleaners. AB - Attempts have been made to justify the sexual division of labor by referring to women's relative physical weakness, lack of technical competence, and dislike for dirty work. An analysis of the work activity of train cleaners in France showed a rigid sexual division of labor, in which the task of toilet cleaning was exclusively assigned to women. Interviews and observations revealed a number of physical constraints associated with the work, and particularly with toilet cleaning. Women employees suffered from many musculoskeletal problems and were frequently absent from work. Toilet cleaning involved traveling distances of over 20 kilometers per day and maintaining uncomfortable postures. Twenty-five percent of time during the actual cleaning was spent in a crouched position. The work was complex, involving a number of choices and decisions, as well as requiring a variety of techniques adapted to the differences in the nature and location of soil. This "women's job" was dirty and physically demanding, and required technical skill. Among train cleaners, the inability to rotate jobs due to sex typed job assignment may be associated with specific health and safety risks for both sexes. PMID- 8425783 TI - Pharmaceuticals, patents, and politics: Canada and Bill C-22. AB - In response to high drug prices, the Canadian government amended the country's patent act in 1969 to allow for compulsory licensing to import pharmaceuticals. As a result of the legislation, by 1983 drug costs in Canada were over $200 million lower than they would otherwise have been. The multinational drug industry was strongly opposed to compulsory licensing, despite any evidence that its economic position had been harmed. Restoration of patent protection for drugs was one of the key U.S. demands during free-trade negotiations between Canada and the United States in 1985-1987. The result was Bill C-22, which gave new drugs protection from compulsory licensing for seven to ten years. This article analyzes the impact of Bill C-22 on the generic industry, the creation of jobs in research and development, drug prices, and research and development expenditures. It concludes with an examination of future demands from the pharmaceutical industry. PMID- 8425784 TI - Evaluation of the national cancer program and proposed reforms. AB - A statement by 68 prominent national experts in cancer prevention, carcinogenesis, epidemiology, and public health, released at a February 4, 1992, press conference in Washington, D.C., charged that the National Cancer Institute (NCI) has misled and confused the public by repeated claims of winning the war against cancer. In fact, age-standardized incidence rates have escalated to epidemic proportions over recent decades, while the ability to treat and cure most cancers has not materially improved. Furthermore, the NCI has minimized evidence for increasing cancer rates, which are largely attributed to smoking, trivializing the importance of occupational carcinogens as non-smoking attributable causes of lung and other cancers, and to diet per se, in spite of tenuous and inconsistent evidence and ignoring the important role of carcinogenic dietary contaminants. Reflecting this near exclusionary blame-the-victim theory of cancer causation, with lockstep support from the American Cancer Society and industry, the NCI discounts the role of avoidable involuntary exposures to industrial carcinogens in air, water, food, the home, and the workplace. The NCI has also failed to provide any scientific guidance to Congress and regulatory agencies on fundament principles of carcinogenesis and epidemiology, and on the critical needs to reduce avoidable exposures to environmental and occupational carcinogens. Analysis of the +2 billion NCI budget, in spite of fiscal and semantic manipulation, reveals minimal allocations for research on primary cancer prevention, and for occupational cancer, which receives only +19 million annually, 1 percent of NCI's total budget. Problems of professional mindsets in the NCI leadership, fixation on diagnosis, treatment, and basic research, much of questionable relevance, and the neglect of cancer prevention, are exemplified by the composition of the National Cancer Advisory Board. Contrary to the explicit mandate of the National Cancer Act, the Board is devoid of members authoritative in occupational and environmental carcinogenesis. These problems are further compounded by institutionalized conflicts of interest reflected in the composition of past executive President's Cancer Panels, and of the current Board of Overseers of the Sloan-Kettering Memorial Cancer Center, the NCI's prototype comprehensive cancer center, with their closely interlocking financial interests with the cancer drug and other industries. Drastic reforms of NCI policies and priorities are long overdue. Implementation of such reforms is, however, unlikely in the absence of further support from industrial medicine professionals, besides action by Congress and concerned citizen groups. PMID- 8425785 TI - Advertisements for medicines in leading medical journals in 18 countries: a 12 month survey of information content and standards. AB - The information content of 6,710 advertisements for medicines in medical journals was surveyed to provide a baseline for monitoring the effect of WHO's Ethical Criteria for Medicinal Drug Promotion. The advertisements (ads) appeared during 12 months (1987-1988) in 23 leading national medical journals in 18 countries. Local participants, mostly doctors or pharmacists, examined them. The presence or absence in each ad of important information was noted. In most ads the generic name appeared in smaller type than the brand name. Indications were mentioned more often than the negative effects of medicines. The ads gave less pharmacological than medical information. However, important warnings and precautions were missing in half, and side effects and contraindications in about 40 percent. Prices tended to be given only in countries where a social security system pays for the medicines. The information content of ads in the developing countries differed surprisingly little from that in the industrialized countries. Almost all the ads (96 percent) included one or more pictures; 58 percent of these were considered irrelevant. The authors believe it is a mistake to regard ads as trivial. If they are not considered seriously they will influence the use of medicines as they are intended to do, but read critically they can provide useful information. PMID- 8425786 TI - Problems in distribution of scientific knowledge: intrauterine contraceptive devices and drug catalogs. AB - Intrauterine contraceptive devices (IUDs) are a popular method of contraception worldwide. However, some serious problems have been associated with them. Finland has developed and now manufactures and exports IUDs. Therefore, drug control and the quality of drug information existing in Finland are significant for other countries, as well. This study analyzes the information in the Finnish commercial drug catalog on copper-releasing IUDs and compares it with the scientific literature, the instructions from the licensing authority, and material in its U.S. counterpart, during the last two decades. The results indicate that the distribution of scientific knowledge to the drug catalogs has often been slow. In the early 1980s Finnish manufacturers did not give any practical information on their products, and then and later the Finnish catalog was less comprehensive than the U.S. catalog. The variations in the control system in different nations were reflected in the contents of the Finnish catalog. For practitioners, drug catalogs are important sources of medical information. The results of this study demonstrate (1) that more attention should be paid to the contents of these catalogs, and (2) the continuous need for up-to-date, unbiased drug information. PMID- 8425787 TI - Market reforms in Swedish health care: a threat to or salvation for the universalistic welfare state? AB - This report presents the main characteristics of reforms in the Swedish health services, as exemplified by the "Stockholm Model" introduced in 1992 in Stockholm county. The author discusses the motives behind these reforms, the already evident increases in costs that are occurring, and the effect of these reforms on public support for the welfare state. PMID- 8425788 TI - Commentary on Canadian health insurance: lessons for the United States. AB - The Government Accounting Office's comparatively favorable report on Canada's National Health Insurance program (Medicare) prompted a firestorm of reaction: criticism from the health insurance industry primarily and praise from advocates of single-payer models of American reform particularly. Congressional hearings aired this controversy, and this article is a revised version of the author's testimony to the Government Operations Committee, June 18, 1991. The author examines the legitimacy of cross-national comparison as a general analytic tool and the lessons to be learned from North American health care comparisons in particular. In the final section he critically discusses two sets of myths about Canada's experience with universal health insurance: those regarding the desirability of the Canadian system itself and those questioning the transplantability (adaptability) of the model to the United States. PMID- 8425789 TI - Further refinements of Canadian/U.S. health cost containment measures. AB - Critics of the Canadian health care system have argued that the lower health care share of gross national product (GNP) in Canada relative to the United States is more likely to be associated with a relatively more rapid growth in GNP in Canada than with the ability of the Canadian single-payer system to contain costs. In this article the authors use both the level and the average annual growth rate of health care's share of GNP to provide an assessment of cost containment for the United States and Canada. They conclude that the suggestion that the success of the Canadian system has been an illusion created by its more rapid growth in GNP is not supported once the appropriate adjustments are made to the data. PMID- 8425790 TI - What's blocking health care reform? PMID- 8425791 TI - Unhealthy money: the growth in health PACs' congressional campaign contributions. AB - The health and insurance industries have launched a massive lobbying and public relations effort to undermine popular support in the United States for a national health care system. As part of their campaign, these industries have increased dramatically their PAC contributions to members of Congress. This article provides a detailed account of these PAC contributions to congressional incumbents and candidates, and discusses additional information on the health insurance industry's advertising campaigns. PMID- 8425792 TI - George Bush's ruling class. PMID- 8425793 TI - Skin signs and internal malignancies. PMID- 8425794 TI - The Fine page: risk of cancer in patients with dermatomyositis. PMID- 8425795 TI - Evaluation of manuscripts reporting results of clinical drug trials. PMID- 8425796 TI - Urinary retention associated with herpes zoster infection. AB - BACKGROUND: Herpes zoster infection particularly involving the sacral dermatomes has been associated with bladder and bowel dysfunction, most commonly urinary retention. CASE REPORTS: We report two patients who developed acute urinary retention, one of whom also had constipation, within days of herpes zoster skin lesions of the S2-S4 dermatomes. CONCLUSIONS: Herpes zoster is a reversible cause of neurogenic bladder and bowel dysfunction and should be considered in a patient that presents with acute urinary retention and/or constipation. Sensory abnormalities and flaccid detrusor paralysis are most likely involved in the pathogenesis. PMID- 8425797 TI - Tongue involvement in lepromatous leprosy. AB - BACKGROUND: Involvement of the oral cavity in lepromatous leprosy is well documented. The tongue may demonstrate multiple nodules, thickening, and scarring. METHODS: Ten consecutive untreated patients with lepromatous leprosy with a bacteriologic index of 4+ or more were clinically and histopathologically studied for evidence of tongue involvement. RESULTS: Three patients showed clinical tongue involvement, as a nodulo-plaque lesion in one patient and fissured tongue (lingua plicata) in two patients. Tongue was clinically normal or showed nonspecific changes in the remaining seven patients. Histologic evidence of tongue involvement by lepromatous process was seen in six patients, including three without clinical involvement. CONCLUSIONS: We conclude from this study that the tongue is as prone to involvement by lepromatous process as buccal and palatal mucosa. PMID- 8425798 TI - Pediculosis capitis in Istanbul. AB - BACKGROUND: Pediculosis capitis is a worldwide problem. We studied the prevalence of this infestation in Istanbul. METHODS: We reviewed the records of the Dermatology Clinic of Istanbul University Cerrahpasa Medical Faculty from a 20 year period (1970-1989). RESULTS: The number of cases of pediculosis capitis have gradually increased over this period, with a peak in 1981. Women are more involved than men. There has been a significant increase in cases in children of school and pre-school ages. CONCLUSIONS: Many factors may have contributed to this increase: poor hygiene and socioeconomic conditions, lack of medical training, and probably some resistance to pediculicide preparations in the lice. The stigma attached to the disease may prevent people from coming forward. PMID- 8425799 TI - Clinical and laboratory evaluation of AIDS trichopathy. AB - BACKGROUND: The present study reports the incidence of trichocutaneous disorders studied in 500 patients infected with HIV 1 in a large university-based setting. Correlation of these findings with immunologic function at the time of diagnosis is presented. Unusual presentations and therapeutic interventions are discussed. Prognosis as related to various trichocutaneous disorders is elaborated. METHODS: All patients in this study were HIV 1 positive by Western blot assay. T-cell subsets were evaluated by monoclonal antibodies against T-cell surface markers. Hair disorders were analyzed by means of light hair pull test, hair mount, polarizing microscopy, trichogram, scalp biopsy, and cultures for bacteria, fungi, and mycobacteria as indicated. Trichologic manifestations were classified based upon immunologic correlation of absolute level of helper T cells/mm3. RESULTS: The majority of hair disorders in the study population occurred with helper T cell numbers of less than 150/mm3. Papulosquamous problems including seborrheic dermatitis and psoriasis were most commonly noted followed by disorders of cell growth cycle regulation and trichokeratinization, i.e., telogen effluvium and loose anagen syndrome. CONCLUSIONS: Multiple trichocutaneous disorders occur in the setting of retroviral infection. Most of these disorders occur in the setting of progressive immunoincompetence. The awareness of the disorders described here will aid the clinician in both the recognition and management of pilar aberrations in the appropriate clinical setting. PMID- 8425800 TI - Tinea capitis in Saudi Arabia. AB - BACKGROUND: Tinea capitis is a dermatophyte infection of the scalp, eyebrows, and eyelashes caused by species of Microsporum and Trichophyton. The purpose of this study was to discover the incidence and causal agents of tinea capitis in Saudi Arabia. METHODS: Hair roots, skin scrapings, and pus swabs were collected from patients clinically diagnosed with tinea capitis and were processed for fungus. RESULTS: Of 372 patients with tinea capitis investigated in Saudi Arabia, 240 were found to be positive by direct microscopic examination, and the causal agent was isolated from 237 patients. Tinea capitis accounted for 47.7% of all superficial mycoses, and 97% of it occurred in children below 15 years of age. Inflammatory lesions were found in 35% of cases, and 10 of them presented with kerion celsi. Favus-type lesion was found in one. Microsporum canis was the most common etiologic agent, responsible for 82.3% of the infections. Trichophyton violaceum was the next most common agent (13.9%), followed by M. audouini (2.2%); T. mentagrophytes, T. rubrum, T. verrucosum, and T. simii were isolated from one patient each. This is the first report of T. simii infection in a Saudi man. CONCLUSIONS: Although none of the patients owned pets, the predominance of M. canis may be explained by the large number of cats in the neighborhood. The disappearance of favus due to T. schoenleinii may be due to improved socioeconomic conditions. Our results agreed with two previous reports. PMID- 8425801 TI - Malignant melanoma in children. AB - BACKGROUND: While significant risk factors for malignant melanoma may initially develop or are first seen in childhood, the actual occurrence of this neoplasm in prepubertal children is uncommon. METHODS: A retrospective study of malignant melanoma in Puerto Ricans up to 16 years of age occurring from 1973 to 1990 was carried out by identifying those cases in the Puerto Rico Cancer Registry. RESULTS: A total of seven cases were found consisting of three boys and four girls with ages ranging from 22 months to 16 years and comprising 0.94% of the total melanomas. In three of the seven cases, there was a history of a previously existent small congenital melanocytic nevus on the area. Three cases were Clark's level I, two level II, and in two cases with proved metastatic disease, Clark's level of invasion were not reported. Those cases with Clark's level I and II had a 100% 5-year survival. CONCLUSIONS: Although rare, malignant melanoma in children can be as aggressive as in adults. Among the known factors predisposing to malignant melanoma, three out of seven cases developed within a small congenital nevus, two of which occurred during the first decade of life. Due to the rarity of this event in our population, it appears unreasonable to excise all small congenital nevi during the first decade of life. Even for those who advocate excision of all small congenital nevi, the evidence at present suggests that such small nevi very rarely undergo malignant change before puberty and therefore a policy of observation in childhood and offering excision around the time of puberty is perfectly logical. PMID- 8425802 TI - Detection of early lesions of "ungual" malignant melanoma. AB - BACKGROUND: The nail area is commonly affected by malignant melanoma. The prognosis of malignant melanoma of the nail is poor, because at the time of diagnosis most lesions are in the advanced stage. Correct diagnosis of early lesions could improve the prognosis. METHODS: For 3 years, all patients with nail pigmentation at the dermatology clinic were screened for five specific criteria for the diagnosis of early lesions of malignant melanoma. Histologic examination was performed on 10 of 29 lesions. RESULTS: Five of the 29 lesions were advanced malignant melanoma, easily diagnosed clinically. Two of the remaining 24 lesions fulfilled most of our clinical criteria of early malignant melanoma of the nail apparatus; that is, they appeared as melanonychia striata during adulthood, were wide in breadth measuring 9 and 11 mm, and showed variegated shades of brown. Periungual pigmented macule (Hutchinson's sign) was observed in one of the two cases. Total resection of the lesions was performed, followed by skin grafting. CONCLUSIONS: Histologically, an increased number of atypical melanocytes, mainly arranged as solitary units, were observed only in the epithelia of the nail matrix and of the nail-bed, confirming that these lesions were "ungual" malignant melanoma in situ. Such an early lesion of malignant melanoma of the nail apparatus can be completely cured with conservative excision, and the phalanx of the affected digit can be preserved. PMID- 8425803 TI - Paraneoplastic pemphigoid-pemphigus? Subepidermal bullous disease with pemphigus like direct immunofluorescence. AB - BACKGROUND: Paraneoplastic pemphigus is a recently described variant of pemphigus, in which classic immunologic features of pemphigus are associated with subepidermal erythema multiforme-like bullae. CASE REPORTS: Two patients with underlying malignant neoplasm had a bullous disease, which had clinical and histologic features suggestive of bullous pemphigoid while direct immunofluorescence revealed intercellular deposits of IgG and C3 throughout the malpighian layer of epidermis, a pattern usually diagnostic for pemphigus. Indirect immunofluorescence studies were negative. One mg/kg/day prednisone was enough to cure the eruption in a couple of weeks without any recurrence. CONCLUSIONS: The disease differs from the recently reported paraneoplastic pemphigus and from other bullous diseases associating circulating pemphigus-like antibodies with clinical, histologic, and often even immunologic features of bullous pemphigoid. PMID- 8425804 TI - Familial aplasia cutis congenita associated with limb anomalies and tetralogy of Fallot. PMID- 8425805 TI - Lichen planus of the nails presenting as trachyonychia. PMID- 8425806 TI - Hypertrichosis lanuginosa acquista associated with florid cutaneous papillomatosis. PMID- 8425807 TI - Erythema annulare centrifugum as the presenting sign of Hodgkin's disease. PMID- 8425808 TI - Wound healing. PMID- 8425809 TI - Treatment of rosacea: topical clindamycin versus oral tetracycline. AB - BACKGROUND: A new topical antibiotic preparation, clindamycin in a lotion base, was compared with oral tetracycline in the treatment of rosacea. Forty-three patients clinically diagnosed as having rosacea were examined in an investigator blinded study. METHODS: Patients used topical clindamycin lotion applied twice daily or the usual oral dose of tetracycline hydrochloride (250 mg four times a day for 3 weeks, then 250 mg twice a day for the remaining 9 weeks). Patients' lesions were examined clinically at 3-week intervals over a period of 12 weeks. RESULTS: Topical clindamycin treatment produced similar clinical results to oral tetracycline and was superior in the eradication of pustules. CONCLUSIONS: These results show topical clindamycin in a lotion base to be a safe and effective alternative to oral tetracycline therapy in the treatment of rosacea. PMID- 8425810 TI - Effect of topical paromomycin on cell-mediated immunity during cutaneous leishmaniasis. AB - BACKGROUND: The aim of this study was to evaluate the effect of topical paromomycin treatment on the development of immunity during cutaneous leishmaniasis. METHODS: Three parameters of immunity were measured in the course of the disease: leishmanicidal effector activity, lymphocyte proliferation (cell mediated immunity), and antibody levels (humoral immunity). Peripheral blood specimens of 55 treated and 36 untreated patients were tested. RESULTS: The main results of this study showed that there was a significant delay in the development of leishmanicidal effector activity and to a lesser extent also a delay in the development of antigen-specific proliferative response in the treated compared with the untreated group. No difference was observed between the groups regarding the values achieved in the various tests. CONCLUSIONS: These results suggest that topical paromomycin treatment delays the development of cell mediated immunity but does not affect the levels of immunity that are eventually achieved. PMID- 8425811 TI - Decline of German dermatovenereology under the Nazi regime. PMID- 8425812 TI - Toxic epidermal necrolysis following Yersinia enterocolitica infection. PMID- 8425813 TI - Linear porokeratosis. PMID- 8425814 TI - Discoid lupus erythematosus of lower lip and heterotopic salivary glands. PMID- 8425815 TI - Oral lesions of lichen planus. PMID- 8425816 TI - Proctor Lecture. The function of alpha-crystallin. PMID- 8425817 TI - A method for the isolation of retinal pigment epithelial cells from adult rats. AB - PURPOSE: Isolating retinal pigment epithelium (RPE) cells from adult rat was the goal of this study. METHOD: A modification of the procedure of Mayerson et al was the method employed. After the cornea and lens were removed, the eyecup was treated with hyaluronidase plus collagenase. The neural retina was carefully removed with little RPE attached. The eyecup, free of retina, was treated with the same enzyme cocktail that loosens any attached rod outer segments as well as the attachment of the RPE to Bruch's membrane. RPE cells were isolated in calcium free medium as sheets of cells. RESULTS: Light and electron microscopy revealed good cell morphology. Purity of the RPE preparation was established by microscopy, polyacrylamide gel electrophoresis, and lipid analysis. The viability of the isolated RPE was about 74% by trypan blue exclusion test. CONCLUSIONS: This technique enables RPE to be isolated from adult rat in quantities sufficient for biochemical analysis. PMID- 8425818 TI - Assessing the reliability, discriminative ability, and validity of disability glare tests. AB - PURPOSE: To gather information regarding the reliability, discriminative ability, and validity of disability glare tests. METHODS: The following glare tests were evaluated: the Miller-Nadler, Vistech MCT8000, Berkeley, van den Berg Straylightmeter, and the Brightness Acuity Tester used with the Pelli-Robson and Regan charts. Three test evaluation criteria were used: (1) repeatability- comparing test scores on two visits; (2) discriminative ability--the tests' ability to differentiate between young and old subjects and between old normal and cataract subjects; (3) validity--comparing cataract test scores with the reference standard of the van den Berg Straylightmeter. Three subject groups were evaluated: young normals (n = 24, mean age 24.3 +/- 3.3 yr), older normals (n = 22, mean age 66.0 +/- 6.2 yr), and early cataract (n = 33, mean age 70.6 +/- 8.1 yr). RESULTS AND CONCLUSIONS: Data indicate that contrast sensitivity or low contrast acuity measured in the presence of glare are superior to disability glare scores in assessing cataract patients with normal neural function. Under glare conditions, contrast sensitivity and low contrast acuity scores from the Pelli-Robson, Regan, and Berkeley tests provide similarly reliable, discriminative, and valid measures of visual assessment in cataract. The Miller Nadler glare tester poorly detects and measures subtle changes in the ocular media, such as early cataract, because of its large step sizes at low contrast thresholds. The poor reliability of the Vistech MCT8000 limits its usefulness. The study suggests that unless good chart design and psychophysics are used, the geometry and intensity of the glare source are of little importance. PMID- 8425819 TI - On the statistical reliability of letter-chart visual acuity measurements. AB - PURPOSE: To determine boundary values of test-retest reliability and sensitivity to acuity change that are unlikely to be exceeded in any clinical situation, for a popular visual acuity chart (Lighthouse/ETDRS) with three scoring methods; to discuss general methodological issues associated with statistical accuracy of optotype chart testing; and to link measures of test reliability to measures of sensitivity to change. METHODS: Five highly practiced subjects were tested using a computer-controlled acuity testing system in a procedure designed to reduce measurement error. Subjects read the computerized chart 156 times, yielding a sample of 78 test-retest comparisons. RESULTS: Under conditions likely to minimize variability, visual acuity may, with 95% confidence, be ascertained only within +/- 0.1 log units, using this chart with the recommended letter-by-letter scoring. Detecting a significant change in visual acuity requires about +/- 0.14 log units for the same degree of confidence. CONCLUSIONS: These measurements may be viewed as approaching the upper limit of reliability of this letter chart. Reliability probably is considerably less in typical usage. PMID- 8425820 TI - Quantitating nuclear opacification in color Scheimpflug photographs. AB - PURPOSE: To test the validity of the color component subtraction method, a new objective technique of measuring nuclear cataract, and to correlate the tangent values of the different degrees of nuclear opacities to their corresponding subjective nuclear cataract grading using the Lens Opacities Classification System II and to another objective method of measuring nuclear cataract. METHODS: Densitometries for red, green, and blue colors of the color Scheimpflug image were conducted simultaneously along the anteroposterior axis of the lens nucleus. The three color curves were subtracted from each other, giving rise to three subtracted curves: blue-red, green-blue, and red-green. This technique was applied to 99 color Scheimpflug photographs taken in 99 eyes of 51 patients with varying degrees of nuclear opacities. Using linear regression analysis, the regression coefficient obtained, called tangent value (TV), represents the degree of nuclear opacification. RESULTS: Higher tangent value indicated greater nuclear opacification and was associated with higher correlation coefficient. This was seen graphically as an increased steepness in the slope of the subtracted curve. Good correlation existed between the subjective nuclear opacity grading and the objective tangent value. When the tangent values were correlated to the mean density of the three original color curves, good correlation was present in red and green, but only fair for blue, where maximum light scattering occurs. CONCLUSIONS: These results support the validity of color component subtraction technique in measuring nuclear cataract. This technique provides quantitative measures of nuclear opacification, has good reproducibility, and is useful for monitoring nuclear cataract longitudinally. PMID- 8425821 TI - Characterization of FN-C/H-V, a novel synthetic peptide from fibronectin that promotes rabbit corneal epithelial cell adhesion, spreading, and motility. AB - PURPOSE: Fibronectin promotes corneal epithelial cell adhesion and motility in vitro and plays an important role in corneal re-epithelialization during corneal wound healing. Multiple domains contribute to the adhesion- and motility promoting activity of fibronectin. The aim of this study was to identify amino acid sequences that contribute to the rabbit corneal epithelial (RCE) cell adhesion- and motility-promoting activity of the 33 and 66 kD carboxy-terminal heparin-binding fragments of fibronectin. METHODS: Synthetic peptides derived from the 33/66 kD fragments of fibronectin were tested for their ability to directly promote RCE cell adhesion, spreading, and motility. To assess the contribution of these peptides to the activity of fibronectin and the 33/66 kD fragments of fibronectin, synthetic peptides, and antibodies against these peptides were tested for their ability to block RCE cell adhesion, spreading, and motility. RESULTS: In this study, we identified a novel peptide sequence derived from the 33/66 kD fragments of fibronectin, FN-C/H-V (WQPPRARI), that directly promotes the adhesion, spreading, and migration of RCE cells in a concentration dependent manner. A second peptide from the 33/66 kD fragments of fibronectin, FN C/H-IV (SPPRRARVT), promoted RCE cell adhesion and spreading, but did not promote RCE cell migration. In contrast, two synthetic peptides from the 33/66 kD fragments of fibronectin that were previously shown to promote RCE cell adhesion (FN-C/H-I and FN-C/H-III) did not promote RCE cell spreading or migration. Soluble FN-C/H-V inhibited RCE cell adhesion to surfaces coated with FN-C/H-V, the 33/66 kD fragments of fibronectin, and to fibronectin. In addition, polyclonal anti-FN-C/H-V IgG inhibited RCE cell adhesion to FN-C/H-V, the 33/66 kD fragments of fibronectin, and to fibronectin. Finally, polyclonal anti-FN-C/H V IgG also inhibited RCE cell haptotactic migration on the 33/66 kD fragments. CONCLUSIONS: These data suggest that the amino acid sequence defined by peptide FN-C/H-V contributes to the adhesion-, spreading-, and motility-promoting activity of the 33/66 kD carboxy-terminal heparin-binding fragments of fibronectin. Given the important role of fibronectin in corneal wound healing, these findings provide additional insight into the complex molecular basis of corneal epithelial cell interactions with fibronectin and may be important in the context of corneal wound healing. PMID- 8425822 TI - Fibrin induction of thrombospondin in corneal endothelial cells in vitro. AB - PURPOSE: Fibrin deposition in the anterior chamber of the eye occurs in response to injury or inflammation and can permanently damage the corneal endothelium. Fibrin functions as a mediator of inflammation and wound healing by affecting cell morphology and function in a variety of cells, including corneal endothelial cells. We hypothesized that fibrin can directly induce corneal endothelial cells to express injury-related proteins (eg, thrombospondin [TSP]) necessary for corneal repair processes. METHODS: Bovine corneal endothelial cells (BCECs) were pulse- or continuously labeled with 35S-methionine in the presence or absence of in situ polymerized fibrin (2 mg/ml). BCECs were harvested after 3-48 hr, and 35S labeled proteins were analyzed by SDS-PAGE, autoradiography, and immunochemical techniques. RESULTS: Fibrin selectively induced BCECs to express a high molecular weight (MW) protein that was present extracellularly in conditioned medium and fibrin matrix. This induction represented a 3-5 fold increase relative to nonfibrin-treated BCECs, was not accompanied by corresponding changes in 35S labeled intracellular proteins, and was evident at early (3 hr) or late times (24 hr) post-fibrin treatment. The induced protein had an apparent MW of 180 kD (reduced) and > 420 kD (nonreduced), consistent with the characteristics of TSP. A polyclonal antibody to human TSP recognized the reduced form (180 kD) on Western blots and the native form (> 420 kD) in immunoprecipitation studies. CONCLUSIONS: Fibrin induces BCECs to express TSP, a matrix protein involved in cell-cell and cell-matrix interactions and implicated in wound healing. PMID- 8425823 TI - Recovery of photoreceptor outer segment length and analysis of membrane assembly rates in regenerating primate photoreceptor outer segments. AB - PURPOSE: Photoreceptor outer segments are in a dynamic state of membrane addition and disposal. This study was undertaken to determine how a standardized period of retinal detachment and varying periods of reattachment affect the renewal process. METHODS: To investigate the effects that retinal detachment and reattachment may have on this process, the neural retina from 12 adult rhesus monkeys (Macaca mulatta) was detached from the overlying retinal pigment epithelium (RPE) by subretinal injection of a balanced salt solution. After a standardized detachment period of 7 days, the two tissue layers were reapposed. Animals were labeled with 3H-fucose and killed at times ranging from 3-150 days after reattachment. RESULTS: During the 7 day detachment period, the majority of rod outer segments (ROS) and cone outer segments (COS) degenerated, but inner segments remained intact. During the first week after reattachment, a rapid increase in rod and cone outer segment length occurred in the absence of disc shedding. This was accompanied by re-establishment of a modified morphologic relationship between the apical processes of the RPE and the regenerating outer segments. ROS and COS regained approximately 40% of their control lengths after a 2 wk reattachment period. By 30 days of reattachment, ROS had regained 72% of their normal length and COS had regained approximately 48%. After 150 days of reattachment, photoreceptor outer segment mean length was not statistically different from control areas. Autoradiographic results confirmed that new disc membranes were synthesized after reattachment. The rate of ROS membrane assembly was subnormal at reattachment time points up to 30 days. CONCLUSIONS: Retinal detachment leads to a reduction in photoreceptor outer segment absolute length and membrane assembly rates. Increasing time of retinal reattachment is positively correlated with an increase in outer segment absolute length and a corresponding increase in membrane assembly rates. This recovery pattern in eyes without underlying pathology and after a relatively brief detachment interval may represent the upper limit of the recovery process. PMID- 8425825 TI - Static accommodation in congenital nystagmus. AB - PURPOSE: To conduct a comparative study of static accommodative function between individuals with normal vision (n = 10) and patients with congenital nystagmus (n = 12). METHODS: The component contribution to monocular steady-state accommodation (slope of the accommodative stimulus/response function, accommodative controller gain, tonic accommodation, and depth-of-focus) was assessed subjectively using a Hartinger coincidence-optometer, except for depth of-focus, which was determined psychophysically. RESULTS: The group mean slope for the patients with nystagmus was not significantly different from that found in the normal subjects. However, their variability was markedly increased. Therefore, the patients with nystagmus were divided into three subgroups with regard to the normal accommodative stimulus/response function slope criterion. The majority of patients with nystagmus (n = 10) exhibited slopes that were outside of normal limits, being greater than (n = 4) or less than (n = 6) the normal range. Depth-of-focus was the only parameter found to be significantly different between the normal and the nystagmus groups. When the nystagmus group was divided with respect to etiology--ie, albinotic (n = 4) versus idiopathic (n = 8)--there were no significant differences found for the various accommodative parameters. CONCLUSIONS: We speculate that the primary component contributing to the anomalous accommodative behavior was the increased depth-of-focus, with this perhaps being related to abnormal fixational eye movements and eccentric fixation, and more generally related to overall reduced sensitivity resulting from the early abnormal visual experience. PMID- 8425824 TI - Accelerated heavy particles and the lens. VII: The cataractogenic potential of 450 MeV/amu iron ions. AB - PURPOSE: To determine the cataractogenic potential dose of high velocity iron ions as a fixation of dose administered singly or fractionated. The dose is critical to risk assessment and to theories of radiation action and cataractogenesis. METHODS: Twenty-eight-day-old rats were examined by slit-lamp biomicroscopy on a weekly-bi-weekly basis for more than 2 yr after radiation exposure. For the acute exposure study doses of 1, 2, 5, 25, and 50 cGy were evaluated. The fractionated regimens involved total doses of 2, 25, and 50 cGy. The reference radiation consisted of 50, 100, 200, or 700 cGy of 250 kilovolt (peak) x-rays. RESULTS: In accordance with previous findings in the rat using 570 MeV/amu 40Ar ions, the relative biologic effectiveness increased rapidly with decreasing dose, reaching values as high as 100. Unlike 40Ar ions, fractionation of the 56Fe doses did not produce a consistent enhancement at any of the doses examined. CONCLUSIONS: The data support the previous findings of a high cataractogenic potential for high linear energy transfer (LET) radiation. The effectiveness for the production of cataracts increases with decreasing dose relative to x-rays and is independent of dose protraction. Although the present study did not reveal a consistent enhancement of effect when the ions were applied in fractions, the results are consistent with at least one theory of the inverse dose-rate effect observed for high-LET radiation. PMID- 8425826 TI - Atropine reduces experimental myopia and eye enlargement via a nonaccommodative mechanism. AB - PURPOSE: To determine whether the muscarinic antagonist atropine effectively reduces or prevents experimentally induced myopia via a nonaccommodative mechanism. METHODS: Chicks were monocularly deprived (MD) of pattern vision by placement of a translucent occluder over the left eye. In two of the three MD groups, chicks received a series of intravitreal injections of atropine (n = 8) or saline vehicle (n = 8) with MD. Control groups (n = 8) of chicks were employed to assess the effects of MD, intravitreal injections, and drug effects. RESULTS: In sham-injected or saline-injected MD chicks, 8 days of MD produced -18.5 D and 20.9 D of experimental myopia, respectively. In atropine-injected MD chicks, 8 days of MD produced only -2.8 D of experimental myopia. This significant reduction in experimentally induced myopia in atropine-injected MD chicks was associated with a marked reduction in the relative axial elongation of the deprived eye (0.21 mm) when compared to saline-injected or sham-injected MD chicks (1.04 mm and 1.00 mm). This reduction in axial length in atropine-injected MD chicks was predominantly the result of a reduction in vitreous chamber elongation, although a reduction in anterior segment depth also was observed. Mean equatorial diameter was significantly reduced in atropine-injected MD chicks compared to saline-injected and sham-injected MD chicks, although to a lesser extent. Control experiments demonstrated that intravitreally injected atropine did not reduce carbachol-induced accommodation or light-induced pupil constriction in the skeletal intraocular muscles of the chick eye. CONCLUSIONS: These findings demonstrate that chronic administration of the muscarinic antagonist atropine prevents experimentally induced myopia in chick via a nonaccommodative mechanism. PMID- 8425827 TI - Effect of protein kinase C inhibitors and activators on corneal re epithelialization in the rat. AB - PURPOSE: To examine the ability of protein kinase C (PKC) inhibitors and activators to influence the rate of corneal re-epithelialization in the rat. METHOD: Rat corneas with 3 mm diameter central epithelial abrasions were organ cultured in control medium or in medium with inhibitors or activators of PKC. RESULTS: In control corneas, the defect was completely re-epithelialized by 25 hr. In the presence of the PKC inhibitors staurosporine (100 nM), sphinganine (50 mumol/l), or H-7 (100 mumol/l) there were significantly larger epithelial defects than in controls after 5-25 hr of incubation. Re-epithelialization rates were similar to control corneas when the incubation medium contained HA1004 (100 mumol/l), an analogue of H-7 that is a potent inhibitor of cyclic adenosine monophosphate- and cyclic guanosine monophosphate-dependent protein kinases and a weak inhibitor of PKC. Two PKC activators, 1-oleoyl-2-acetyl-sn-glycerol (OAG) and phorbol 12-myristate 13-acetate (PMA), were unable to enhance the rate of epithelial wound healing. CONCLUSIONS: Our results suggest that PKC activity is an important factor in regulating corneal epithelial wound healing, presumably by influencing cell migration. Moreover, the results with OAG and PMA suggest that PKC is maximally activated during re-epithelialization in this organ-culture assay. PMID- 8425828 TI - Spontaneous corneal neovascularization in nude mice. Local imbalance between angiogenic and anti-angiogenic factors. AB - PURPOSE: The present study considered the hypothesis that spontaneous neovascularization of the cornea of nude mice results from a local imbalance between angiogenic and anti-angiogenic factors. METHODS: The presence of angiogenic and anti-angiogenic factors was revealed by exchanging orthotopic corneal allografts between nude BALB/c mice and normal (hirsute) euthymic BALB/c mice and observing the presence, intensity, and degree of corneal neovascularization before and after grafting. RESULTS: Avascular corneal grafts from normal BALB/c donors resisted neovascularization after grafting to spontaneously vascularized graft beds in nude mice. In contrast, spontaneously vascularized corneal grafts from nude mice remained vascularized over 2 mo after grafting to similar nude recipients. Although corneal grafts from nude donors stimulated neovascularization in normal BALB/c recipients, most of the vessels regressed by the 6th week post transplantation. CONCLUSIONS: The results confirm that there is an imbalance between angiogenic and anti-angiogenic factors in the nude mouse cornea. The cornea of the nude mouse displays more angiogenic activity and less anti-angiogenic activity than that of the normal mouse. Most angiogenic activity of the nude mouse cornea appears to reside in the epithelium. PMID- 8425829 TI - Immunolocalization of TGF-beta 1, TGF-beta 2, and TGF-beta 3 in the anterior segment of the human eye. AB - PURPOSE: To determine the distribution of transforming growth factor type beta (TGF-beta) in the anterior segment of the human eye. This knowledge is important because TGF-beta may regulate various physiologic responses in the anterior segment by controlling cell proliferation and differentiation, angiogenesis, and extracellular matrix composition. METHODS: Immunohistochemical methods were used to localize the beta 1, beta 2, and beta 3 isoforms of TGF-beta in the anterior segment of the human eye. RESULTS: Eight of eight eyes (six eye bank specimens and two eyes enucleated because of choroidal melanoma) exhibited staining for at least one of the TGF-beta isoforms. TGF-beta 1 was found in superficial limbal epithelial cells (four of eight eyes) and in the stroma proximal to the ciliary processes (seven of eight eyes). TGF-beta 2 was found in superficial limbal epithelial cells (six of eight eyes), the conjunctival stroma (eight of eight eyes), in the ciliary processes (three of eight eyes), and in a diffuse distribution in the region of the radial and circular muscles of the ciliary body (eight of eight eyes). In addition, TGF-beta 2 was found in the stroma adjacent to the pigmented epithelium in the pars plana (eight of eight eyes). TGF-beta 3 was found in white blood cells in one of eight specimens; otherwise it was not found in the anterior segment. The corneal stroma, corneal endothelium, trabecular meshwork, iris, and ciliary epithelia did not exhibit immunoreactivity with the antibodies used in this study. CONCLUSION: TGF-beta 1 and TGF-beta 2 have a distinct and specific distribution in the anterior segment of the adult human eye. PMID- 8425830 TI - Foveal cone involvement in retinitis pigmentosa progression assessed through flash-on-flash parameters. AB - PURPOSE: To compare psychophysical Naka-Rushton parameters in retinitis pigmentosa (RP) patients and healthy controls using a flash-on-flash increment threshold paradigm, and to measure changes of these parameters with RP progression. METHODS: Sixty-six RP patients and 10 normal subjects were tested, and their maximum response (Rmax), half-saturation intensity (sigma), and slope (n) parameters were estimated. RESULTS/CONCLUSIONS: Rmax in RP patients is decreased significantly with respect to the range in normal controls and continues to decrease (0.024 log units/yr) with disease progression. The distribution of sigma in RP patients differs from that in normal subjects, showing lower values in general, but no progression. Small differences in parameter distributions among genetic or pathophysiologic RP subcategorizations were found, but these do not fulfill stricter statistical criteria required for multiple comparisons. Measurement noise, inherent in the flash-on-flash paradigm, exert considerable influence on the quality of the data, as was demonstrated through repeated measures and a Monte Carlo simulation. PMID- 8425831 TI - Foveal cone involvement in retinitis pigmentosa progression assessed through psychophysical impulse response parameters. AB - PURPOSE: To compare psychophysical impulse response parameters in retinitis pigmentosa (RP) patients and healthy controls using a temporal contrast sensitivity threshold paradigm, and to measure changes in these parameters with RP progression. METHODS: Sixty-six RP patients and 10 healthy control subjects were tested, and the amplitude and timing parameters of the psychophysical impulse response function were computed through time-domain transformation under assumption of minimum-phase properties. RESULTS/CONCLUSIONS: The initial rise time of the impulse response, ta, was significantly lengthened in RP patients compared to controls, as was the fall time from peak to trough, tp-->t. The log peak-to-trough amplitude, log R, was significantly reduced. With disease progression, all impulse response parameters continued to move away from the normal range. Only minor distinctions according to RP pathophysiologic subtype or mode of inheritance were found, supporting the hypothesis of a common course of the secondary retinal degeneration across different RP subcategories. PMID- 8425832 TI - The role of mast cells in endotoxin-induced uveitis. AB - PURPOSE: To investigate the role of mast cells in the induction of endotoxin induced uveitis (EIU). METHODS: We previously showed that the mean mast cell number in the anterior uvea was greatest for Lewis rats, lowest for Brown Norway (BN) rats, and intermediate for LBNF1 rats. In the current experiment, we assessed the time of onset and severity of EIU in these three rat strains. We then studied changes in mast cell numbers in the anterior uvea during the induction period of EIU. RESULTS: Time of onset and severity of EIU were related to the number of mast cells in the anterior uvea. EIU occurred earliest in the Lewis rats, and the maximum mean grade of ocular inflammation on a scale of 0 (no inflammation) to 4 (severe inflammation) +/- standard error of the mean was 3.87 +/- 0.13 for Lewis rats, 1.06 +/- 0.06 for BN rats, and 1.19 +/- 0.12 for LBNF1 rats. The difference between the mean grade of inflammation in the Lewis rats and the other two strains was highly statistically significant (P < .001). In the Lewis rats, mast cell numbers +/- SEM decreased from 68.9 +/- 10.8 at baseline to 49.6 +/- 5.9 4 hr after lipopolysaccharide (LPS) injection and to 27.6 +/- 8.4 8 hr after LPS injection, suggesting that mast cell degranulation occurs before the development of EIU. CONCLUSION: Mast cells in the anterior uvea appear to potentiate endotoxin-induced ocular inflammation. PMID- 8425833 TI - Trabecular cells express receptors that bind TGF-beta 1 and TGF-beta 2: a qualitative and quantitative characterization. AB - PURPOSE: To quantitate the receptors for transforming growth factor (TGF)-beta 1 on trabecular cells in culture and to determine the relative affinities of TGF beta 1 and TGF-beta 2 for these receptors. METHODS: We quantitated the receptors for TGF-beta 1 by Scatchard analysis of radioligand binding of 125I-TGF-beta 1 to cultured porcine trabecular cells. We established the relative affinities of TGF beta 1 and TGF-beta 2 for the receptors by competitive binding of 125I-TGF-beta 1 with increasing concentrations of the unlabeled TGF-beta 1 or TGF-beta 2. We also investigated the binding of 125I-TGF-beta 1 after pre-treatment of trabecular cells with heparinase. RESULTS: Trabecular cells expressed approximately 4,000 high-affinity receptors per cell for TGF-beta 1, with a dissociation constant (Kd) of 15.8 +/- 7.6 pmol/l. By varying the concentrations of the unlabeled growth factors, we determined that the relative affinities of TGF-beta 1 and TGF beta 2 for the receptors were 16 pmol/l and 50 pmol/l, respectively. Heparinase treatment of the trabecular cells did not change the binding affinity of the receptor for 125I-TGF-beta 1. CONCLUSIONS: Our findings show that trabecular cells express heparinase-insensitive TGF-beta receptors that have an approximately threefold greater affinity for TGF-beta 1 than for TGF-beta 2. Based on the present investigation, together with our previous data on the molecular weights of the binding sites, we conclude that trabecular cells do possess types II and III receptors but not type I receptors. PMID- 8425834 TI - Aging studies on normal lens using the Scheimpflug slit-lamp camera. AB - PURPOSE: To study the changes in density and thickness in normal lenses related to aging, and to study changes in anterior chamber depth related to aging. METHODS: Eighty nine normal volunteers (ages 9-80 yr) were examined and their eyes were photographed to obtain Scheimpflug photographs. The images were digitized and linear densitometry was performed, dividing the lens into five areas: posterior capsular (area 1), posterior cortical (area 2), nuclear (area 3), anterior cortical (area 4), and anterior capsular (area 5). Total lens thickness and anterior chamber depth were similarly measured for 90 normal eyes from the densitometry profiles. These were correlated with age. RESULTS: There was a strong positive correlation between increasing age and the density in all lens areas (area 2: r = 0.805; P < 0.0001; area 3: r = 0.836, P < 0.0001; area 4: r = 0.767, P < 0.0001; and area 5: r = 0.319, P < 0.0023), except the posterior capsular area, where correlation was negative (area 1: r = -0.426; P < 0.0001). In addition, there was a significant correlation between age and overall lens thickness (r = 0.756; P < 0.0001), thickness of nucleus (r = 0.543; P < 0.0001), and cortex (r = 0.632; P < 0.0001), and a negative correlation with anterior chamber depth (r = -0.513, P < 0.0001). CONCLUSION: This report shows human lens changes in density and thickness correlated with aging using Scheimpflug photography and image analysis techniques. The results will aid future development of systems for automated detection, classification, and monitoring of human cataracts, as well as other anterior segment disorders. PMID- 8425836 TI - Oxygen-induced retinopathy. PMID- 8425835 TI - The effects of acetazolamide on visual function in retinitis pigmentosa. AB - PURPOSE: To study the effects of acetazolamide on central and peripheral visual function in patients with retinitis pigmentosa (RP) who showed no evidence of macular edema. METHODS: Thirteen patients with retinitis pigmentosa participated in a preliminary study. Measures of central and peripheral visual function were obtained before and after an 8 wk period on acetazolamide. An additional 10 patients participated in a cross-over study. They were placed on a placebo for an 8 wk period, then on acetazolamide for a second 8 wk period. RESULTS: None of the patients in the preliminary study showed significant changes in visual acuity, color vision, foveal cone pathway sensitivities, focal electroretinogram (ERG) amplitudes, or in any ERG parameter. Three patients, however, showed significant changes in visual field area and in dark-adapted thresholds. None of the patients in the cross-over study showed significant increases in visual field area. CONCLUSIONS: Given the results and the reports of side-effects, it is difficult to justify using acetazolamide to improve retinal function in RP patients who show no evidence of cystoid macular edema. PMID- 8425837 TI - Behavioral perimetry in monkeys. AB - PURPOSE: Normative data on the systematic changes in visual sensitivity as a function of retinal eccentricity have provided the basis for efficient threshold strategies and data analysis routines for static perimetry. The standard methods of assessing visual field changes in patients also could be used for monkeys with experimentally induced ocular disorders if the normal visual fields of monkeys and humans were similar. METHODS: Normal visual field data from three rhesus monkeys were compared to data from eight human subjects using the standard threshold programs of the Humphrey Field Analyzer. RESULTS: The experimental paradigm developed for these measurements provided excellent behavioral control for the monkeys, with reliability indices well within acceptable limits. The visual field data from monkeys were comparable to those from humans with respect to: (1) sensitivity as a function of stimulus field size; (2) the derived Statpac global indices; and (3) the variance of threshold measurements across the visual field. CONCLUSION: The visual fields of monkeys and humans are similar, and the techniques of computerized perimetry may be applied to monkey subjects without significant modification. PMID- 8425838 TI - Mechanisms of histamine-induced relaxation in external and internal ophthalmic arteries. AB - PURPOSE: Mechanisms that underlie the relaxant response to histamine were examined in dog external (a branch of external carotid artery) and internal (a branch of internal carotid artery) ophthalmic arteries (EOA and IOA). METHODS: Changes in isometric tensions were recorded in helical strips of the arteries with and without the endothelium. RESULTS: Histamine predominantly produced relaxations in EOA and IOA, partially contracted with prostaglandin (PG) F2 alpha. The relaxation of IOA almost was abolished by treatment with cimetidine (10(-5) mol/l), whereas the response of EOA was partially attenuated by treatment with cimetidine or chlorpheniramine (10(-6) mol/l) and was abolished with their combined treatment. Endothelium denudation depressed the relaxation in EOA but did not affect the response of IOA. The response to histamine of EOA was inhibited by treatment with indomethacin (10(-6) mol/l) or tranylcypromine (10( 4) mol/l), a PGI2 synthesis inhibitor, only when the endothelium was present, but additional treatment with chlorpheniramine did not further inhibit relaxation. On the other hand, IOA's response to histamine was not inhibited by indomethacin, despite the presence of endothelium. CONCLUSIONS: The histamine-induced relaxation in EOA may be associated with the release of vasodilator PGI2 through the activation of H1 receptors in the endothelium and with the direct action on H2 receptors in smooth muscle, whereas the relaxation in IOA is mediated exclusively by H2 receptors in smooth muscle. PMID- 8425839 TI - Oxygen reactivity of the feline isolated ophthalmociliary artery. AB - PURPOSE: The goal of this study was to investigate the modulating role of oxygen tension on noradrenaline (NA) and KCl responses in the ophthalmociliary artery and to ascertain whether these effects are mediated by the endothelial cells. METHODS: The isometric tension generated by myograph ring segments activated by NA or KCl was measured as the PO2 of the bathing solution was decreased in discrete steps from 506.1 +/- 16.0 mmHg to 29.4 +/- 1.4 mmHg in preparations with and without endothelial cells. RESULTS: Vessels pre-activated with K+ Krebs solution were insensitive to oxygen tensions between 506.1 +/- 16.0 mmHg and 124.6 +/- 4.2 mmHg. Lower PO2s caused a graded and increasing contraction that reached 176 +/- 12% of the contraction at the highest PO2. Vessels pre-activated with NA had a dichotomous response to reductions in oxygen tension: 44% of vessels showed a graded contraction, whereas a graded relaxation was observed for the remaining 56% of vessels as bath PO2 was reduced. In all cases, a functional endothelium was demonstrated. However, deliberate disruption of the endothelium caused all vessels pre-activated with NA to exhibit a consistent graded contraction for PO2s below 124.6 +/- 4.2 mmHg, similar to that observed for endothelial intact vessels pre-activated with K+ Krebs. The acetylcholine dose response curve and passive tension were not affected by changes in oxygen tension. CONCLUSIONS: Endothelial cells modify the intrinsic smooth muscle response to a gradual reduction in PO2 by releasing relaxing and contracting factors, causing the observed dichotomous response in NA-activated vessels. However, the KCl-induced response is modulated only by low oxygen tensions. PMID- 8425840 TI - Ocular ischemia and the effects of allopurinol on functional recovery in the retina of the arterially perfused cat eye. AB - PURPOSE: This study sought to examine the acute effects of ocular ischemia and reperfusion on retinal function and determine the extent to which recovery during reperfusion is improved by allopurinol (AP), a blocker of xanthine oxidase (XO). The latter is presumed to be a major factor in the formation of free radicals associated with reperfusion of ischemic tissue. METHODS: Electroretinographic (ERG) responses were recorded simultaneously from the two isolated, arterially perfused eyes obtained from the same cat. One eye served as the control and received only the standard perfusate; the other eye was infused with AP before and after a 3 hr period of total ischemia. RESULTS: After the prolonged period of nonperfusion, recovery of the electroretinographic components was incomplete to varying degrees. Maximum b-wave amplitudes recovered only to 17 +/- 5% (mean +/- SEM) of baseline, whereas the a-wave grew to 60 +/- 10% of its baseline value. For both measures, the recovery of electroretinographic amplitude was significantly greater in AP-treated eyes than in the control eyes. CONCLUSION: Generation of free radicals by XO contributes to the retinal damage and loss of function that occurs after a period of ischemia and subsequent reperfusion. PMID- 8425841 TI - Proctor Award and Lecture. Introduction of Joseph Horwitz for the Proctor Medal. PMID- 8425842 TI - Retinal hemodynamics in proliferative diabetic retinopathy. A laser Doppler velocimetry study. AB - PURPOSE: This study investigated retinal hemodynamic changes associated with different pathologic features observed on fundus color and fluorescein angiography in patients with proliferative diabetic retinopathy. METHODS: Retinal circulatory characteristics were investigated in 25 eyes of 23 diabetic patients with proliferative retinopathy using a combination of bidirectional laser Doppler velocimetry and monochromatic fundus photography. RESULTS: Eyes with severe capillary nonperfusion had 32% less average volumetric blood flow rate (Q) than eyes with less severe nonperfusion (P = 0.0005). In addition, eyes with severe vessel staining with fluorescein had 20% less average Q than eyes without staining (P = 0.0508). Eyes with severe fluorescein leakage in the macula had a 17% larger total venous cross-section than eyes with milder leakage (P = 0.027). Eyes with clinically significant macular edema had 11% larger average venous diameter than eyes without this feature (P = 0.0085). CONCLUSIONS: Severe capillary nonperfusion and vessel staining with fluorescein are associated with decreased retinal blood flow rates. Vasodilatation may be an important factor for increased vascular permeability and macular edema in proliferative diabetic retinopathy. PMID- 8425843 TI - Neuroprotectants in Honghua: glucose attenuates retinal ischemic damage. AB - PURPOSE: This study examined the neuroprotective properties of Honghua, an extract of safflower used as an herbal medicine in China, in several experimental models of retinal ischemia. METHODS: Honghua and other agents were tested (1) in the ex vivo chick embryo retina assay (CER) for anti-excitotoxin efficacy and against simulated ischemia (30 min glucose/oxygen deprivation); and (2) in the in vivo adult rat retina dye-photothrombosis assay. Active components of Honghua were purified by conventional chromatographic techniques. RESULTS: In the CER, Honghua protected against excitotoxicity of glutamate, N-methyl-D-aspartate, kainate, and quisqualate, and against neuronal degeneration caused by simulated ischemia. Honghua more potently protected against simulated ischemia than against the agonists. In the in vivo adult rat retina, ischemic damage was reduced greatly by intravitreal injection of Honghua. An approximately 100-fold purification of an active principle was achieved chromatographically. The purest fractions were rich in glucose, so the effects of glucose in the ischemia models were determined. Many neuroprotective effects of Honghua were mimicked by pure solutions of equivalent glucose concentration. Glucose (> 3.2 mmol/l) in the CER ischemia assay provided protection. Glucose did not protect against the lesions induced by direct application of the excitotoxic agonists. Intravitreal injection of glucose provided highly significant neuroprotection in the adult rat retina dye-photothrombosis model. CONCLUSIONS: These results suggest that retinal excitotoxic damage in vivo can occur secondary to depletion of cellular energy reserves, and therefore may be prevented by simple procedures that maintain the availability of energy sources. PMID- 8425844 TI - Recoverin, but not visinin, is an autoantigen in the human retina identified with a cancer-associated retinopathy. AB - PURPOSE: We investigated the hypothesis that visinin, a cone-specific protein first characterized in chicken retina, is a cone homologue of recoverin and may be the cancer-associated retinopathy (CAR) autoantigen in human cone cells. METHODS: Visinin was purified from chicken retinas and tested for binding by CAR antisera. In addition, antibodies specific to visinin were used immunocytochemically and for Western analysis to determine whether visinin is present in human or bovine retinas. Anti-peptide antibodies against recoverin were used immunocytochemically to localize recoverin to mammalian cone cells. RESULTS: CAR antisera recognized recoverin but not visinin. Furthermore, visinin could not be detected in mammalian retinas by immunocytochemical methods or by attempts to purify the protein. In contrast to visinin, antibodies specific for different regions of the recoverin molecule stained both rod and cone cells in the human retina. CONCLUSIONS: Visinin is not the CAR autoantigen in human cone cells. Differences between recoverin and visinin probably reflect species differences rather than rod-cone differences. Recoverin, or a nearly identical molecule, is present in mammalian cones and likely is the cone cell CAR autoantigen. PMID- 8425845 TI - Autoantibodies against retinal bipolar cells in cutaneous melanoma-associated retinopathy. AB - PURPOSE: This study's goal was to determine the pathophysiology of the retinopathy that occurs in patients with metastatic cutaneous melanoma and sudden onset of night blindness, the so-called melanoma-associated retinopathy (MAR) syndrome. We tested the hypothesis that sera from two MAR patients contained autoantibodies that reacted with "on" bipolar cells of the human retina. METHODS: Immunofluorescence was performed on cryostat sections of unfixed normal human retinas. Sera and IgG fractions were tested from the two MAR patients and 38 control subjects (28 patients with metastatic melanoma, but no visual symptoms; two patients with non-MAR retinopathy; and eight normal subjects). RESULTS: The sera and IgG fractions from both MAR patients but from none of the control subjects produced heavy immunostaining of bipolar cells, which were identified as rod bipolars by a double labeling procedure using anti-protein kinase C. CONCLUSIONS: We hypothesize that MAR patients generate autoantibodies against a melanoma antigen that cross react with bipolar cells of the retina. These antibodies, by an unknown mechanism, may cause abnormalities of the rod and cone systems that are characteristic of MAR. PMID- 8425846 TI - Detection and differentiation of diffuse liver disease by quantitative echography. A retrospective assessment. AB - RATIONALE AND OBJECTIVES: The detectability of diffuse liver diseases by quantitative echography was retrospectively investigated using scans of patients with known pathologic findings (n = 103) and of normal subjects (n = 129). The authors determined the best set of quantitative parameters for this task. METHODS: Quantitative echography was comprised of acoustospectrographic parameters (frequency dependence of attenuation and backscattering) and image texture parameters. The disease processes studied included: acute hepatitis, hepatitis/cirrhosis, alcoholic hepatitis/cirrhosis, primary biliary cirrhosis, and steatosis. RESULTS: Correct differentiation of these diseases ranged from 88% to 97%. Correlations between histologic grading and echographic parameters were poor. With only one exception, the differentiation between any two of the diseases could be made in 60% to 99% of cases. Different parameters better differentiated abnormal from normal scans than among diseases. CONCLUSIONS: The detection of diffuse liver diseases can be based on echographic parameters, related to a diffuse scattering model, whereas the differentiation among diseases needs additional parameters derived from a structural scattering model. Further studies are indicated to assess the prospective potential of the devised methods. PMID- 8425847 TI - Cardiovascular responses after ionic and nonionic magnetic resonance contrast media in rats with acute myocardial infarction. AB - RATIONALE AND OBJECTIVES: Contrast media may have quantitatively or even qualitatively different effects in the presence of underlying pathologic states compared with normal states. This study was designed to examine and compare the hemodynamic effects of bolus administration of ionic (gadopentetate dimeglumine) and nonionic (gadodiamide) magnetic resonance (MR) contrast media in rats subjected to acute myocardial infarction. METHODS: Acute myocardial infarction was induced in two groups of rats (n = 20) by ligating the left coronary artery. Each animal received four bolus injections, iso-osmolar glucose followed by three incremental doses of either an ionic or a nonionic MR contrast agent (0.1, 0.3, and 0.5 mmol/kg). The effects of iso-osmolar glucose and each dose of MR contrast agent on the cardiovascular system were monitored for 15 minutes. RESULTS: Iso osmolar glucose injection did not cause hemodynamic parameters to significantly differ from baseline values. Nonionic gadodiamide produced no significant hemodynamic effects at all injected doses compared with iso-osmolar glucose. However, ionic gadopentetate dimeglumine caused significant deleterious hemodynamic effects in a dose-dependent fashion. Gadopentetate dimeglumine caused depression in left ventricular (LV) systolic pressure and systemic arterial pressure at the lowest dose (0.1 mmol/kg). At the maximum dose (0.5 mmol/kg), gadopentetate dimeglumine decreased systolic arterial pressure by 48%, rate pressure product by 55%, LV end systolic pressure by 48%, rate of rise of LV pressure (dP/dt) by 55%, and heart rate by 10%. LV end diastolic pressure increased by 46%. Arrhythmias were observed in 20% (2/10) of the animals after injection of gadopentetate dimeglumine, but not after gadodiamide. CONCLUSIONS: Compared with ionic gadopentetate dimeglumine, nonionic gadodiamide is a hemodynamically safe MR contrast agent in this experimental model when it is injected as a rapid bolus at high doses and in the presence of acute myocardial infarction. PMID- 8425848 TI - Sodium-calcium balance and cardiac function with isotonic iodixanol. An experimental study in the isolated rat heart. AB - RATIONALE AND OBJECTIVES: Three formulations of the nonionic dimer iodixanol (150/200/300 mg I/mL), made isotonic by the addition of NaCl (70/53/24 mM), were investigated regarding their potential for depressing cardiac contractility during coronary angiography. To maintain a stable cardiac function, the authors sought the requirements for the addition of a balanced amount of calcium (Ca). METHODS: Iodixanol 150, 200, and 300 mg I/mL were applied as a short-lasting bolus in isolated perfused rat hearts in the absence and presence of added Ca. The contractile function was assessed by measurement of changes in left ventricular developed pressure (LVDP). RESULTS: A transient LVDP depression was markedly alleviated by adding 0.2 to 0.4 mM Ca and almost abolished by 0.4 to 0.9 mM. Ca higher than 0.9 mM led to unstable hearts and too-extensive Ca loading. CONCLUSIONS: Caution should be used when adding Ca to iodixanol, particularly with sodium-calcium (Na-Ca) relationships. Appropriate Ca concentrations are probably 0.6 mM for iodixanol (150 mg I/mL); 0.4 to 0.6 mM for iodixanol (200 mg I/mL; and 0.4 mM for iodixanol (300 mg I/mL). PMID- 8425849 TI - Gadolinium-ethoxybenzyl-DTPA, a new liver-directed magnetic resonance contrast agent. Absence of acute hepatotoxic, cardiovascular, or immunogenic effects. AB - RATIONALE AND OBJECTIVES: Gadolinium-ethoxybenzyl-DTPA (Gd-EOB-DTPA) is a recently introduced experimental magnetic resonance (MR) contrast agent for hepatic imaging. Although liver enhancement has been investigated in a number of animal models, tolerance evaluations of Gd-EOB-DTPA injection have been limited. METHODS: The authors investigated acute hepatotoxicity in an isolated perfused rat liver model, cardiovascular effects in the anesthetized rat, and potential immunogenicity of Gd-EOB-DTPA using detection of specific antibodies. RESULTS: Using perfused rat liver model, no significant deviation could be observed for functional parameters, liver enzymes, or potassium release, comparing Gd-EOB-DTPA to a control, but there was a significant choleresis (+250% bile flow). Hemodynamic effects of Gd-EOB-DTPA were observed after femoral bolus injection, but only with relatively high dosages (0.3-0.5 mmol/kg, 10-fold the likely clinical dose in humans). Experimental conditions, idealized for antibody induction, failed to cause an IgG immune response to Gd-EOB-DTPA in the intact rat. CONCLUSIONS: The results further support preliminary conclusions that Gd-EOB DTPA is a well-tolerated MR contrast agent. PMID- 8425850 TI - Necrotic areas in VX2 carcinoma of rabbits. Correlation of magnetic resonance imaging and pathologic appearance. AB - RATIONALE AND OBJECTIVES: To assess the potential of magnetic resonance imaging (MRI) for evaluating the evolution of tumor necrosis, the authors evaluated 28 VX2 carcinomas inoculated in the thigh muscles of rabbits. METHODS: MRI of VX2 carcinomas at 2.0 T was done with spin-echo technique 1 week (14 tumors) and 3 weeks (14 tumors) after the inoculation of tumor cells. T1- and T2-weighted images were obtained with 500/30 and 2,500/80 sequences; the authors also performed enhanced T1-weighted images with contrast medium. MRI and histopathologic findings were compared. RESULTS: Enhanced T1-weighted images with gadolinium-DTPA (Gd-DTPA) (dimeglumine gadopentetate) could depict necrosis in almost all tumors (27/28) and were the most sensitive technique followed by T2- and T1-weighted images. T2-weighted images best permitted the evaluation of the characteristics of necrotic areas in VX2 carcinomas, followed by T1-weighted images with contrast enhancement, and unenhanced T1-weighted images. On T2 weighted images, low-intensity areas of necrotic areas correspond to areas of early necrosis, isointense areas corresponded to areas of intermediate necrosis, and high-intensity areas corresponded to the late phase of necrosis. CONCLUSIONS: MRI is useful in depicting necrosis and characterizing different phases of necrosis in VX2 carcinomas. PMID- 8425851 TI - Computed tomography and histologic results in the early stages of endotoxin injured pig lungs as a model for adult respiratory distress syndrome. AB - RATIONALE AND OBJECTIVES: To determine early radiographic changes in diffuse alveolar injury, the authors correlated computed tomography (CT) and histopathology in pigs with recurrent endotoxinemia. METHODS: Five pigs received recurrent endotoxin over a 17-hour period. Three pigs received physiologic saline and served as controls. Hemodynamic and blood-gas data were analyzed. CT was performed immediately before killing the animals. The lungs were cut into 5-mm thick slices in the same axis as the CT scans and were investigated by light and electron microscopy. RESULTS: Hemodynamic data, blood-gas analysis, and morphologic changes closely simulated the clinical situation of septic shock in the five pigs that had received endotoxin. Results of histologic examination depicted changes similar to those associated with adult respiratory distress syndrome (ARDS). CT clearly demonstrated both interstitial, and to a minor degree, intra-alveolar lesions in the endotoxin-injected group, which correlated well with dilated lymph vessels, thickened interstitium, and areas of dystelectasis on histologic examination. Although there was a rather uniform clinical picture, CT and histologic findings showed different degrees of involvement. CONCLUSIONS: CT clearly depicts changes in endotoxin-injured pig lungs in an early clinical state, which are similar to changes associated with ARDS on histologic examination. PMID- 8425852 TI - An animal model of corpus callosum impingement as seen in patients with normal pressure hydrocephalus. AB - RATIONALE AND OBJECTIVES: Callosal impingement has been postulated to cause the symptoms associated with normal pressure hydrocephalus. The authors developed an animal model for the investigation of corpus callosum impingement by the falx cerebri. METHODS: The corpus callosum was compressed from above by a plastic blade and surgically placed in the interhemispheric fissure in adult Sprague Dawley rats. Magnetic resonance imaging (MRI) was performed both preoperatively and postoperatively. The brains of the rats also were examined grossly at the time of autopsy. RESULTS: All rats survived the surgical procedure. MRI proved to be a suitable method to image the corpus callosum, to confirm the location of the blade, and to demonstrate the corpus callosum impingement. MRI correlated well with necroscopy sections. CONCLUSIONS: It was possible to surgically produce callosal impingement in rats, and this impingement could be confirmed by MRI. In the future, this rat model of callosal impingement will be used to search for evidence of changes in metabolism, neuroelectrical activity, behavior, and neuronal anatomy which are known or are thought to be associated with hydrocephalus. PMID- 8425853 TI - Roentgen and the "new light"--Roentgen's moment of discovery. Part 2: The first glimmer of the "new light". PMID- 8425854 TI - Quantification of failure to demonstrate statistical significance. The usefulness of confidence intervals. PMID- 8425855 TI - In vivo functional imaging using receptor-binding radiopharmaceuticals. Technetium 99m-galactosyl-neoglycoalbumin as a model. PMID- 8425856 TI - Effect of focal zone on a curved array transducer. AB - RATIONALE AND OBJECTIVES: This study investigates the effect of electronic focusing of a linear array transducer on received ultrasound signal. METHODS: Two tissue-mimicking phantoms of known attenuation were imaged with a focal distance that was varied from 2 to 12 cm. Received signal was processed by Fourier analysis to compute average frequency and power as functions of depth within the phantom. RESULTS: Average received frequency increases dramatically over the first centimeter from the transducer and then decreases linearly to 8 to 11 cm with a slope proportional to the phantom's attenuation. Average frequency from greater depths decreases more rapidly and is dependent on the focal zone. Received power peaks at approximately the focal depth and decays exponentially beyond that level; the decay constant is proportional to the phantom's attenuation but independent of focusing. CONCLUSIONS: The frequency and power of received ultrasound signal are altered by electronic focusing in ways that may significantly influence both tissue characterization and clinical imaging applications. PMID- 8425857 TI - Mentoring. Applications for the practice of radiology. PMID- 8425858 TI - Toward a theory of hypothesis generation in diagnostic decision making. PMID- 8425859 TI - Spontaneous perinephric hemorrhage: imaging and management. PMID- 8425860 TI - The night stalker effect: quality improvements with a dedicated night-call rotation. AB - RATIONALE AND OBJECTIVES: The authors assessed whether a non-sleep-deprived, second-year diagnostic radiology resident assigned to an after-hours "night stalker" emergency radiology (ER) rotation in an urban university hospital has a measurable impact on the number and clinical significance of "missed" radiologic findings. METHODS: After-hours Emergency Department (ED) radiographs interpreted by radiology residents between January and December 1991 were reviewed daily by ER faculty. Faculty-modified final interpretations were recorded on a worksheet and given to the attending ED physician (EDP). The EDP reviewed and, if indicated, modified clinical dispositions, and categorized missed diagnoses as requiring recall into the following categories: 1 = immediately, 2 = in 24 to 48 hours, 3 = no recall necessary, or 4 = recognized during patient visit by clinicians. Morbidity attributable to "misses" was graded A to C (A, definite; B, possible; C, none). All cases requiring patient recall were evaluated monthly with follow-up information and classified as false-positive, false-negative, or indeterminant. The relative performance of control (traditional "call") and night stalker groups were compared. RESULTS: Of 26,421 on-call examinations in 1991, there were 489 (1.1%) misses, of which 202 (1.4%) were from the June-to-December study group. The control group residents averaged 2.75 hours of sleep per call night. On night stalker days, on-call residents and the night stalker averaged 5.75 and 7.25 hours of sleep daily, respectively. The fractions (and number) of recall assignments of discordant cases for the control and night stalker groups, respectively, were: 1) immediate 48% (23) and 26% (32); 2) within 48 hours 31% (15) and 26% (31); 3) no recall 79% (38) and 36% (43); and, 4) abnormality not missed by EDPs 10% (5) and 12% (15). Morbidity for the control and night stalker groups, respectively, were: 1) 4% and 0%; 2) 31% and 30%; and 3) 65% and 70%. The amount of rework between July and December 1991 spent by the EDPs to re-evaluate cases because of discordant opinions was more than 68 hours, with no significant difference noted between the study groups. Errors were false-negative, 84.9% (415); false-positive, 7% (34); and indeterminant, 8.2% (40). Radiology faculty errors contributed 5.8% (13) of patient recalls (false-positive, 11; false negative, 2). Finally, 58/78 questionnaire respondents believed that service quality had improved. No one believed that the standard of service had been lowered. CONCLUSIONS: A dedicated night-shift ER coverage of a busy urban ED improves quality, appropriateness, and timeliness of patient care. PMID- 8425861 TI - Results of the 1992 survey of the American Association of Academic Chief Residents in Radiology. AB - RATIONALE AND OBJECTIVES: The authors conducted the annual American Association of Academic Chief Residents in Radiology (A3CR2) survey of chief residents of academic radiology programs to evaluate commonalities and differences among radiology training programs. METHODS: The surveys were mailed to 125 accredited programs in the United States and Canada. The analysis of the returned surveys took into account the geographic region of the respondents. RESULTS: Chief residents from 72 training programs (58%) returned their completed surveys. There are important variations among training programs related to requirements for entry, salary, and night call, and how residents prepare for Board examinations. CONCLUSIONS: The A3CR2 annual survey provides important information for program directors regarding how their training practices might differ from others so that they might better evaluate their programs. PMID- 8425862 TI - Radiology resident selection: results of a survey. AB - RATIONALE AND OBJECTIVES: The author determined which criteria were important to program directors for selecting candidates for diagnostic radiology residency. METHODS: A questionnaire was sent to all radiology residency program directors. They were asked to rank 20 criteria they considered important in a written application. Miscellaneous questions about their program's selection process, including the interview, also were asked. RESULTS: The overall response rate was 69%. Statistical analysis of responses showed significant differences between geographic regions and types of programs (university versus private hospital). Class rank and medical school grades were found to have the highest rating of importance. Nearly half of responding programs use a score sheet to evaluate the written application. On average, almost 60 candidates are invited to interview for approximately five available positions. CONCLUSIONS: Criteria believed to be important for selection and the radiology resident selection process show significant differences among programs. These can be correlated partially with the location and type of program. PMID- 8425863 TI - Step-down units and telemetry monitoring: optimizing utilization. AB - In this article, we address the issue of patient-flow problems in the hospital, which often occur because of improper utilization of step-down (intermediate care) unit resources. Utilization is often adversely affected by the inappropriate use of telemetry monitoring (traditionally offered in step-down units) as a rationale for placing patients in step-down areas, even when they would benefit from other types of monitoring or just the proper level of nursing care. We also discuss the other types of patient monitoring available, as well as other monitoring issues (see "Communicating Alarm Event"). PMID- 8425865 TI - Mispackaging of disposable pressure transducers and continuous-flush devices. PMID- 8425864 TI - Polamedco tracheal tubes. PMID- 8425866 TI - Oxygen concentrators. AB - This study focuses on molecular-sieve concentrators that deliver high-content oxygen for continuous, long-term therapy, which improves and extends the quality of life for adult patients who suffer from such illnesses as chronic obstructive pulmonary disease. We did not include membrane concentrators, which produce a maximum concentration of 40% oxygen (compared with 90% or more for molecular sieve concentrators), because they are not suitable for most long-term oxygen therapy applications. Although oxygen concentrators are most often used at home, they are also occasionally used in hospitals or nursing homes as an economical method of delivering oxygen when built-in systems are not required or available. We evaluated 10 oxygen concentrators from nine manufacturers. All of the units are similar in general design and appearance, except for one 3 L/min capacity unit that is smaller and lighter than the other 4 or 5 L/min units. Our ratings and rankings are based primarily on the availability of an oxygen concentrator status indicator that meets our criteria, as well as on electrical safety and maintenance issues. Nine units are rated Acceptable; one unit is rated Conditionally Acceptable because it poses an electrical safety hazard. Readers are cautioned not to base purchasing decisions on our ratings alone, but on a thorough understanding of all issues surrounding the use of oxygen concentrators, which can be gained only by reading this study in its entirety. In the Discussion section, "Issues in Selecting, Purchasing, and Using Oxygen Concentrators," we discuss important factors to consider, such as required flow, oxygen concentration status indicators, high-temperature performance, electrical safety, noise, maintenance, and cost. Manufacturers' specifications for these and other units are available in ECRI's Hospital Product Comparison System. PMID- 8425867 TI - Flash apertures for improved specimen detail in ultra close-up photography and photomacrography. PMID- 8425868 TI - Seeing and telling. PMID- 8425869 TI - An interview with Clifford L. Freehe, RBP, FBPA. Interview by Dale Tilly. PMID- 8425870 TI - Computer-generated black-and-white tone/line negatives on color reversal film. PMID- 8425871 TI - H. Lou Gibson: a life in photography. PMID- 8425872 TI - Beyond signs and symptoms: the case against a mixed anxiety and depression category. AB - Both primary care physicians and psychiatrists report frequent difficulty in distinguishing between major depression and generalized anxiety disorder. This fact has suggested the need for a new diagnostic category of mixed anxiety and depression. The solution, however, does not address the fundamental issues and creates additional problems. While there are multiple factors contributing to the difficulty in distinguishing between these categories, two are of paramount importance. First, pivotal cross-sectional criteria for the two disorders as enumerated in DSM-III-R are virtually identical. Second, the crux of the distinction between these two disorders involves factors other than the immediate cross-sectional presentation in the office such as the longitudinal course of the patient's complaints. Obtaining such a history takes time and training. For a variety of reasons (e.g., time allocated for patient evaluation, physician training), such a longitudinal history may not be elicited in a primary care office. To respond to these problems by establishing a mixed diagnostic category does not address the primary problems, may encourage cursory evaluations of patients, and will likely hinder research. PMID- 8425873 TI - Mixed anxiety and depression: diagnostic issues. AB - Mixed anxiety and depression (MAD) is a new diagnostic category introduced in the ICD-10 classification for patients seen mainly in primary care settings. These patients are defined as those suffering from symptoms of anxiety and depression of limited and equal intensity accompanied by at least some autonomic features, who do not qualify for specific diagnosis of anxiety or depressive disorders and are independent of stressful life events. The validity of this clinical entity is presently under investigation in the DSM-IV-MAD field trial. Cases of mixed anxiety and depression, however, are not limited to those meeting the criteria of this new "subsyndromal" category. Many patients fulfilling criteria for either depressive or anxiety disorders may also respectively present symptoms, syndromes, or a diagnosis of anxiety or depression. It is still not known whether anxious and depressive symptoms are two different expressions of the same psychopathologic underlying process. Tyrer's recent description of a "general neurotic syndrome" is an attempt to reunify syndromes separated in our present classifications. In this comprehensive approach, anxiety, depression, or MAD states are associated at different times with specific personality features and considered as expressing different levels of overreactivity to various stressful situations. This hypothesis would explain the close relationship existing between these two categories of symptoms and the common efficacy of some psychopharmacologic agents for both anxiety and depressive disorders. PMID- 8425874 TI - Mixed anxiety and depression: clinical implications. AB - Although depressive and anxious symptoms frequently coexist, clinical studies have tended to separate anxiety disorders from depression. A number of developments are now reversing this trend. One of these developments is the reworking of the concept of generalized anxiety disorder (GAD) from that of a residual category (anxiety after all other anxiety disorders are removed) to a generalized anxiety syndrome that includes symptoms of mild depression that are less severe than the symptoms of anxiety. GAD is thereby expanded to a broader concept, namely mixed anxiety/depression (MAD). A second major development, however, posits a different definition of MAD: a stable core of subsyndromal symptoms that do not reach the threshold for the diagnosis of GAD or depression, but which, under stress, will decompensate to an overt anxiety disorder or depression. PMID- 8425875 TI - Comorbidity and mixed anxiety-depressive disorders: is there epidemiologic evidence? AB - Recent epidemiologic studies (i.e., studies conducted since 1980) have consistently demonstrated, on the basis of standardized diagnostic assessments, that there is a substantial overlap between different types of anxiety and depressive disorders. The current literature, however, discusses this issue primarily within the concept of comorbidity and there are some controversies about the existence of a separate disorder of mixed anxiety-depression (MAD). MAD can be defined by the presence of mixed symptoms of depression and anxiety that are below the diagnostic threshold for either one of these diagnoses. Since MAD has not been included in any of the current official classification systems, its prevalence, risk factors, course, and outcome have not been studied specifically in any of the recent epidemiologic studies even though MAD is thought to be very important, especially in primary care settings. This paper reviews recent epidemiologic studies and presents data from the Munich Follow-Up Study, which has found a prevalence of about 1% for MAD as defined by the ICD-10. Despite the lack of clear diagnostic criteria for MAD, there are some indications that: (1) this disorder might be frequent in primary care settings, and (2) patients with MAD frequently demonstrate subjective suffering, show impairment in personal and occupational functioning, and have high health service utilization rates. Current empirical evidence is still insufficient for deciding a suitable classificatory solution for this problem. PMID- 8425876 TI - Marriage, age at last birth and fertility in a traditional Moroccan population. AB - A sample of 842 rural women from Morocco (Amizmiz, Marrakech) was used to examine the relationship between a number of biosocial variables and fertility patterns. For women still in their reproductive years there were significant correlations between family size and woman's age, age at marriage and years of marriage. Among women with completed families, those with early age at marriage ceased childbearing about 10 years before reaching menopause, while women who married later continued to bear children until the end of their fertile life. PMID- 8425877 TI - Genetic distance estimated through surname frequencies of 37 counties from the state of Lara, Venezuela. AB - Genetic distances between all possible pairs of counties (n = 37) in the state of Lara, Venezuela were calculated using surname frequencies and the Euclidean distance as estimator. In general, Euclidean distances were smaller between counties closer together, and the product moment correlation between geographic and Euclidean distances was 0.49 (p < 0.02). The results suggest that, in Lara, geographic distance has been an important determinant of genetic structure, although topography and roadways also have had an important influence upon this structure. PMID- 8425878 TI - Trends in consanguineaous marriage in Karnataka, south India, 1980-89. AB - Analysis of data on 106,848 marriages in the cities of Bangalore and Mysore, South India, between 1980 and 1989 showed that levels of consanguineous marriage varied between cities through time and by religion. The average coefficient of inbreeding was higher in Bangalore (F = 0.0339) than in Mysore (F = 0.0203), principally reflecting large-scale, post-Independence rural migration into Bangalore. Although there was some evidence of a decline in consanguineous marriages in Mysore, there was no convincing support in either city for earlier projections of a rapid reduction in the popularity of unions between close biological relatives. PMID- 8425879 TI - Determinants of coitus after childbirth. AB - Using a sample of 3080 women from Cebu, Philippines, interviewed immediately after childbirth and every 2 months thereafter for 2 years, the determinants of return to coitus are analysed. Eighty per cent of the women returned to coitus before the return of menses, and 90% did so before stopping breast-feeding. In hazards models, variables associated with traditional life styles retarded return to coitus. Situational variables (husband resident, crowding, children aged 6 and under), especially husband residence, and biological variables (mother's age, return to menses, and lactation) were significant predictors of return to coitus. The implications of the analysis for the construction of models of birth interval dynamics are discussed. PMID- 8425880 TI - Breast-feeding patterns in the Philippines: a prospective analysis. AB - Data from the Cebu Longitudinal Health and Nutrition Survey have been used to examine breast-feeding patterns in the Metropolitan Cebu region of the Philippines. The results show that there is extensive and early use of non nutritive liquids, resulting in low levels of exclusive breast-feeding. The use of cross-sectional versus longitudinal data, the definition of exclusive breast feeding, and movements in and out of the exclusively breast-fed category, are considered. PMID- 8425881 TI - Men and family planning in Zambia. AB - This paper examines the sociodemographic factors which influence familiarity with methods of family planning among 85 males holding low paying jobs in the University of Zambia, Lusaka. The results showed that wife's education had a significant and positive effect on husband's familiarity with family planning methods. In the longer term, female education is likely to emerge as an important factor in the onset of fertility decline in Zambia. PMID- 8425882 TI - Do men grow to resemble their wives, or vice versa? AB - Sixty-eight engaged couples, 113 newly weds, and 222 couples married for over 5 years, were studied for a number of physical, psychological and socioeconomic traits. The measurements were repeated a year later. Correlations were highest for age (r = -0.454, 0.903 and 0.888 respectively). They were also high for height (but not weight), neuroticism (but not extroversion), occupation, education, religion and smoking. Similarities were already present about the time of marriage, with little convergence or divergence. Thus, men do not grow to resemble their wives, nor vice versa. PMID- 8425883 TI - Levels and differentials in weight, height and body mass index among mothers in a rural area of Bangladesh. AB - This study examined the variation in weight, height and body mass index of 1048 mothers living in a rural area of Bangladesh in relation to age, education, number of previous pregnancies, number of dead children, religion, family type, family size, and amount of land owned by the household. Multiple regression analysis revealed a positive relationship of education with all three variables. Moslem mothers were on average in better condition than Hindus. The number of dead children showed a negative relationship with height, weight and body mass index. PMID- 8425884 TI - Nuptiality patterns in two west African states. PMID- 8425885 TI - Sterilisation: characteristics of vasectomy acceptors in Delhi. AB - The place of vasectomy within the sterilisation programme in Delhi over the period 1983-88 is reviewed and data on vasectomy acceptance and characteristics of acceptors are analysed. Findings suggest a need to improve the strategy for the promotion of vasectomy within the metropolis. PMID- 8425886 TI - Postpartum tubal sterilisation: an international perspective on some programmatic issues. AB - The demand for postpartum sterilisation (performed within 42 days after delivery), is increasing both in developed and developing countries. The incidence of regret after postpartum sterilisation is important, but it could be minimised by carefully screening risk factors. Using trained paramedical personnel to perform postpartum sterilisation via minilaparotomy where physicians are in short supply appears to be safe and acceptable, under close medical supervision. Including postpartum sterilisation information in the antenatal counselling services effectively strengthens postpartum services and simultaneously helps to minimise subsequent regret. PMID- 8425887 TI - Inbreeding coefficients from the surnames of grandparents of the schoolchildren in Albanian-speaking Italian villages. AB - Data on grandparental surnames were obtained from children in 45 Italo-Albanesi villages in nine provinces of southern Italy and Sicily. Concordance of surnames (isonymy) and inbreedding by village were estimated for each province and on the total sample. Total mean isonymy is 0.0251. The weighted mean inbreeding coefficient, and its random and non-random components are 0.0063, 0.0024 and 0.0039, respectively. Isonymy values are similar to those of rural Italian villages except that Alpine and some Appennine villages appear to be more isolated and inbred. PMID- 8425888 TI - Home-school differences in stress hormone levels in a group of Oxford primary schoolchildren. AB - Twenty-three girls and thirteen boys, aged between 4 and 9 years, gave urine samples which were measured by HPLC for adrenaline, noradrenaline, cortisol and cortisone on a school day and on a home day. Home levels of noradrenaline, cortisol and cortisone were consistently higher than school levels. Cortisol was positively correlated with adrenaline and noradrenaline at home, but not at school. These results may suggest that the school day represents a stimulating but comparatively structured environment with little negative affect, while the home day is characterised by stimulation but relative uncertainty in social interactions. PMID- 8425889 TI - Influence of birth interval and child labour on family energy requirements and dependency ratios in two traditional subsistence economies in Africa. AB - The consequences of different birth intervals on dietary energy requirements and dependency ratios at different stages of the family lifecycle are modelled for Gambian agriculturalists and !Kung hunter-gatherers. Energy requirements reach a peak at between 20 and 30 years after starting a family for the Gambians, and between 15 and 20 years for the !Kung. For the Gambians, shorter birth interval confers no economic advantage over the traditional birth interval of 30 months. For the !Kung, the lack of participation in subsistence activities by children gives an output:input ratio in excess of that reported in other studies, suggesting that they are in a state of chronic energy deficiency. PMID- 8425890 TI - Determinants of low birthweight: a comparative study. AB - The study compares biological, socioeconomic and behavioural determinants of low birthweight in Cameroon and the United States. Some factors in low birthweight are found to be cross-national, but others are specific to the setting. Positive risk factors of low birthweight in both countries include unmarried motherhood, female sex, multiple births, and preterm births. Outcome of the previous pregnancy is a positive risk factor in the US, but not in Cameroon. Significant negative risk factors include prenatal care visits (in both countries), mother's education (in the US only), births to mothers aged 20-34 and birth orders of 2 or more (in Cameroon only). Separate analyses of all births and the subsamples of singleton births reveal that estimates for the two groups differ only marginally. PMID- 8425891 TI - Transvection, nuclear structure, and chromatin proteins. PMID- 8425892 TI - The onset of homologous chromosome pairing during Drosophila melanogaster embryogenesis. AB - We have determined the position within the nucleus of homologous sites of the histone gene cluster in Drosophila melanogaster using in situ hybridization and high-resolution, three-dimensional wide field fluorescence microscopy. A 4.8-kb biotinylated probe for the histone gene repeat, located approximately midway along the short arm of chromosome 2, was hybridized to whole-mount embryos in late syncytial and early cellular blastoderm stages. Our results show that the two homologous histone loci are distinct and separate through all stages of the cell cycle up to nuclear cycle 13. By dramatic contrast, the two homologous clusters were found to colocalize with high frequency during interphase of cycle 14. Concomitant with homolog pairing at cycle 14, both histone loci were also found to move from their position near the midline of the nucleus toward the apical side. This result suggests that coincident with the initiation of zygotic transcription, there is dramatic chromosome and nuclear reorganization between nuclear cycles 13 and 14. PMID- 8425893 TI - Expression of the murine small heat shock proteins hsp 25 and alpha B crystallin in the absence of stress. AB - Stress induces the synthesis of several large and small heat shock proteins (hsp's). Two related small hsp's, hsp25 and alpha B crystallin exist in mice. alpha B crystallin is an abundant protein in several tissues even in the absence of stress. Particularly high amounts accumulate in the eye lens. Here we show that hsp25 is likewise constitutively expressed in many normal adult tissues. In the absence of stress the protein is most abundant in the eye lens, heart, stomach, colon, lung, and bladder. The stress-independent expression pattern of the two small hsp's is distinct. In several tissues the amount of hsp25 exceeds that accumulating in NIH 3T3 fibroblasts in response to heat stress. hsp25, like alpha B crystallin, exists in a highly aggregated form in the eye lens. The expression of hsp25 and alpha B crystallin in normal tissues suggests an essential, but distinct function of the two related proteins under standard physiological conditions. PMID- 8425894 TI - Biosynthesis of glycosylphosphatidylinositol (GPI)-anchored membrane proteins in intact cells: specific amino acid requirements adjacent to the site of cleavage and GPI attachment. AB - Mutational studies were previously carried out at the omega site intact cells (Micanovic, R., L. Gerber, J. Berger, K. Kodukula, and S. Udenfriend. 1990. Proc. Natl. Acad. Sci. USA. 87:157-161; Micanovic R., K. Kodukula, L. Gerber, and S. Udenfriend. 1990. Proc. Natl. Acad. Sci. USA: 87:7939-7943) and at the omega + 1 and omega + 2 sites in a cell-free system (Gerber, L., K. Kodukula, and S. Udenfriend. 1992. J. Biol. Chem. 267:12168-12173) of nascent proteins destined to be processed to a glycosylphosphatidyl-inositol (GPI)-anchored form. We have now mutated the omega + 1 and omega + 2 sites in placental alkaline phosphatase (PLAP) cDNA and transfected the wild-type and mutant cDNAs into COS 7 cells. Only glycine at the omega + 2 site yielded enzymatically active GPI membrane-anchored PLAP in amounts comparable to the wild type (alanine). Serine was less active and threonine and valine yielded very low but significant activity. By contrast the omega + 1 site was promiscuous, with only proline being inactive. These and the previous studies indicate that the omega and omega + 2 sites of a nascent protein are key determinants for recognition by COOH-terminal signal transamidase. Comparisons have been made to specific requirements for substitution at the -1, 3 sites of amino terminal signal peptides for recognition by NH2-terminal signal peptidase and the mechanisms of NH2 and COOH-terminal signaling are compared. PMID- 8425895 TI - Two independent peroxisomal targeting signals in catalase A of Saccharomyces cerevisiae. AB - In contrast to many other peroxisomal proteins catalase A contains at least two peroxisomal targeting signals each sufficient to direct reporter proteins to peroxisomes. One of them resides at the extreme carboxy terminus constituting a new variant of this signal, -SSNSKF, not active in monkey kidney cells (Gould, S. J., G. A. Keller, N. Hosken, J. Wilkinson, and S. Subramani 1989. J. Cell Biol. 108:1657-1664). However, this signal is completely dispensable for import of catalase A itself. In its amino-terminal third this protein contains another peroxisomal targeting signal sufficient to direct reporter proteins into microbodies. This internal signal depends on the context. The nature of this targeting signal might be a short defined sequence or a structural feature recognized by import factors. In addition, we have demonstrated that the carboxy terminal seven amino acids of citrate synthase of Saccharomyces cerevisiae encoded by CIT2 and containing the canonical -SKL represents a targeting signal sufficient to direct reporter proteins to peroxisomes. PMID- 8425896 TI - Cytosol-dependent peroxisomal protein import in a permeabilized cell system. AB - Using streptolysin-O (SLO) we have developed a permeabilized cell system retaining the competence to import proteins into peroxisomes. We used luciferase and albumin conjugated with a peptide ending in the peroxisomal targeting sequence, SKL, to monitor the import of proteins into peroxisomes. After incubation with SLO-permeabilized cells, these exogenous proteins accumulated within catalase-containing vesicles. The import was strictly signal dependent and could be blocked by a 10-fold excess of peptide containing the SKL-targeting signal, while a control peptide did not affect the import. Peroxisomal accumulation of proteins was time and temperature dependent and required ATP hydrolysis. Dissipation of the membrane potential did not alter the import efficiency. GTP-hydrolyzing proteins were not required for peroxisomal protein targeting. Depletion of endogenous cytosol from permeabilized cells abolished the competence to import proteins into peroxisomes but import was reconstituted by the addition of external cytosol. We present evidence that cytosol contains factors with SKL-specific binding sites. The activity of cytosol is insensitive to N-ethylmaleimide (NEM) treatment, while the cells contain NEM-sensitive membrane-bound or associated proteins which are involved in the import machinery. The cytosol dependence and NEM-sensitivity of peroxisomal protein import should facilitate the purification of proteins involved in the import of proteins into peroxisomes. PMID- 8425897 TI - Breaching the diffusion barrier that compartmentalizes the transmembrane glycoprotein CE9 to the posterior-tail plasma membrane domain of the rat spermatozoon. AB - CE9 is a posterior-tail domain-specific integral plasma membrane glycoprotein of the rat testicular spermatozoon. During epididymal maturation, CE9 undergoes endoproteolytic processing and then redistributes into the anterior-tail plasma membrane domain of the spermatozoon (Petruszak, J. A. M., C. L. Nehme, and J. R. Bartles. 1991. J. Cell. Biol. 114:917-927). We have determined the sequence of CE9 and found it to be a Type Ia transmembrane protein identical to the MRC OX-47 T-cell activation antigen, a member of the immunoglobulin superfamily predicted to have two immunoglobulin-related loops and three asparagine-linked glycans disposed extracellularly. Although encoded by a single gene and mRNA in the rat, the majority of spermatozoal CE9 is of smaller apparent molecular mass than its hepatocytic counterpart due to the under-utilization of sites for asparagine linked glycosylation. By fluorescence recovery after photobleaching, CE9 was determined to be mobile within the posterior-tail plasma membrane domain of the living rat testicular spermatozoon, thus implying the existence of a regional barrier to lateral diffusion that is presumed to operate at the level of the annulus. Through the development of an in vitro system, the modification of this diffusion barrier to allow for the subsequent redistribution of CE9 into the anterior-tail domain was found to be a time-, temperature-, and energy-dependent process. PMID- 8425898 TI - Elastic filaments in situ in cardiac muscle: deep-etch replica analysis in combination with selective removal of actin and myosin filaments. AB - To clarify the full picture of the connectin (titin) filament network in situ, we selectively removed actin and myosin filaments from cardiac muscle fibers by gelsolin and potassium acetate treatment, respectively, and observed the residual elastic filament network by deep-etch replica electron microscopy. In the A bands, elastic filaments of uniform diameter (6-7 nm) projecting from the M line ran parallel, and extended into the I bands. At the junction line in the I bands, which may correspond to the N2 line in skeletal muscle, individual elastic filaments branched into two or more thinner strands, which repeatedly joined and branched to reach the Z line. Considering that cardiac muscle lacks nebulin, it is very likely that these elastic filaments were composed predominantly of connectin molecules; indeed, anti-connectin monoclonal antibody specifically stained these elastic filaments. Further, striations of approximately 4 nm, characteristic of isolated connectin molecules, were also observed in the elastic filaments. Taking recent analyses of the structure of isolated connectin molecules into consideration, we concluded that individual connectin molecules stretched between the M and Z lines and that each elastic filament consisted of laterally-associated connectin molecules. Close comparison of these images with the replica images of intact and S1-decorated sarcomeres led us to conclude that, in intact sarcomeres, the elastic filaments were laterally associated with myosin and actin filaments in the A and I bands, respectively. Interestingly, it was shown that the elastic property of connectin filaments was not restricted by their lateral association with actin filaments in intact sarcomeres. Finally, we have proposed a new structural model of the cardiac muscle sarcomere that includes connectin filaments. PMID- 8425899 TI - ptx1, a nonphototactic mutant of Chlamydomonas, lacks control of flagellar dominance. AB - A new mutant strain of Chlamydomonas, ptx1, has been identified which is defective in phototaxis. This strain swims with a rate and straightness of path comparable with that of wild-type cells, and retains the photoshock response. Thus, the mutation does not cause any gross defects in swimming ability or photoreception, and appears to be specific for phototaxis. Calcium is required for phototaxis in wild-type cells, and causes a concentration-dependent shift in flagellar dominance in reactivated, demembranated cell models. ptx1-reactivated models are defective in this calcium-dependent shift in flagellar dominance. This indicates that the mutation affects one or more components of the calcium dependent axonemal regulatory system, and that this system mediates phototaxis. The reduction or absence of two 75-kD axonemal proteins correlates with the nonphototactic phenotype. Axonemal fractionation studies, and analysis of axonemes from mutant strains with known structural defects, failed to reveal the structural localization of the 75-kD proteins within the axoneme. The proteins are not components of the outer dynein arms, two of the three types of inner dynein arms, the radial spokes, or the central pair complex. Because changes in flagellar motility ultimately require the regulation of dynein activity, cell models from mutant strains defective in specific dynein arms were reactivated at various calcium concentrations. Mutants lacking the outer arms, or the I1 or I2 inner dynein arms, retain the wild-type calcium-dependent shift in flagellar dominance. Therefore, none of these arms are the sole mediators of phototaxis. PMID- 8425900 TI - Loss of epithelial differentiation and gain of invasiveness correlates with tyrosine phosphorylation of the E-cadherin/beta-catenin complex in cells transformed with a temperature-sensitive v-SRC gene. AB - Loss of histotypic organization of epithelial cells is a common feature in normal development as well as in the invasion of carcinomas. Here we show that the v-src oncogene is a potent effector of epithelial differentiation and invasiveness. MDCK epithelial cells transformed with a temperature-sensitive mutant of v-src exhibit a strictly epithelial phenotype at the nonpermissive temperature for pp60v-src activity (40.5 degrees C) but rapidly loose cell-to-cell contacts and acquire a fibroblast-like morphology after culture at the permissive temperature (35 degrees C). Furthermore, the invasiveness of the cells into collagen gels or into chick heart fragments was increased at the permissive temperature. The profound effects of v-src on intercellular adhesion were not linked to changes in the levels of expression of the epithelial cell adhesion molecule E-cadherin. Rather, we observed an increase in tyrosine phosphorylation of E-cadherin and, in particular, of the associated protein beta-catenin. These results suggest a mechanism by which v-src counteracts junctional assembly and thereby promotes invasiveness and dedifferentiation of epithelial cells through phosphorylation of the E-cadherin/catenin complex. PMID- 8425901 TI - Adhesion-dependent protein tyrosine phosphorylation in neutrophils treated with tumor necrosis factor. AB - Human neutrophils (PMN) respond to tumor necrosis factor (TNF) by releasing their granules, reorganizing their cytoskeleton, and massively secreting hydrogen peroxide. This response is dependent on adhesion to extracellular matrix proteins and expression of CD11b/CD18 integrins (Nathan, C., S. Srimal, C. Farber, E. Sanchez, L. Kabbash, A. Asch, J. Gailit, and S. D. Wright. 1989. J. Cell Biol. 109:1341-1349). We investigated the role of tyrosine phosphorylation in the response of PMN to TNF. PMN adherent to protein-coated surfaces but not suspended PMN showed tyrosine phosphorylation of several proteins (approximately 150, approximately 115, approximately 75, and approximately 65 kD) in response to TNF. Tyrosine phosphorylation was evident 5 min after addition of TNF and lasted at least 2 h. The tyrosine kinase inhibitors K252a, genistein and ST638 suppressed tyrosine phosphorylation and blocked hydrogen peroxide production in a reversible manner at low concentrations. Tyrosine kinase inhibitors also blocked the spreading of PMN in response to TNF. Dihydrocytochalasin B did not inhibit tyrosine phosphorylation, but in its presence phosphorylation was rapidly reversed. By immunocytochemistry, the majority of tyrosine phosphoproteins were localized to focal adhesions. Thus TNF-induced tyrosine phosphorylation depends on adhesion of PMN to extracellular matrix proteins, and participates in the transduction of the signals that direct the cells to spread on a biological surface and undergo a respiratory burst. PMID- 8425902 TI - Functional characterization of chondroitin sulfate proteoglycans of brain: interactions with neurons and neural cell adhesion molecules. AB - Ng-CAM and N-CAM are cell adhesion molecules (CAMs), and each CAM can bind homophilically as demonstrated by the ability of CAM-coated beads (Covaspheres) to self-aggregate. We have found that the extent of aggregation of Covaspheres coated with either Ng-CAM or N-CAM was strongly inhibited by the intact 1D1 and 3F8 chondroitin sulfate proteoglycans of rat brain, and by the core glycoproteins resulting from chondroitinase treatment of the proteoglycans. Much higher concentrations of rat chondrosarcoma chondroitin sulfate proteoglycan (aggrecan) core proteins had no significant effect in these assays. The 1D1 and 3F8 proteoglycans also inhibited binding of neurons to Ng-CAM when mixtures of these proteins were adsorbed to polystyrene dishes. Direct binding of neurons to the proteoglycan core glycoproteins from brain but not from chondrosarcoma was demonstrated using an assay in which cell-substrate contact was initiated by centrifugation, and neuronal binding to the 1D1 proteoglycans was specifically inhibited by the 1D1 monoclonal antibody. Different forms of the 1D1 proteoglycan have been identified in developing and adult brain. The early postnatal form (neurocan) was found to bind neurons more effectively than the adult proteoglycan, which represents the C-terminal half of the larger neurocan core protein. Our results therefore indicate that certain brain proteoglycans can bind to neurons, and that Ng-CAM and N-CAM may be heterophilic ligands for neurocan and the 3F8 proteoglycan. The ability of these brain proteoglycans to inhibit adhesion of cells to CAMs may be one mechanism to modulate cell adhesion and migration in the nervous system. PMID- 8425903 TI - Content of nonhistone protein in nuclei after hyperthermic treatment. AB - When nuclei were isolated from Chinese hamster ovary cells after being heated, there was a large increase in the amount of 3H-tryptophan labeled nonhistone protein in the nucleus relative to the whole cell. After 15 min or 30 min of heating at 45.5 degrees C, the nuclear nonhistone protein content increased by 1.6 or 1.8, respectively. In contrast, when the nuclear nonhistone protein content was determined in the intact cell by using autoradiography to quantify 3H tryptophan labeled protein in the nucleus and cytoplasm in sections of fixed cells, the nuclear nonhistone protein content increased by only 1.14 or 1.28 for 15 or 30 min at 45.5 degrees C, respectively. Therefore, heat does not induce a massive movement of cytoplasmic protein into the nucleus. PMID- 8425904 TI - Involvement of the Golgi region in the intracellular trafficking of cholera toxin. AB - The intracellular pathway following receptor-mediated endocytosis of cholera toxin was studied using brefeldin A (BFA), which inhibited protein secretion and induced dramatic morphological changes in the Golgi region. In both mouse Y1 adrenal cells and CHO cells, BFA at 1 micrograms/ml caused a 80-90% inhibition of the cholera toxin (CT)-induced elevation of intracellular cAMP. The inhibition of the cytotoxicity of CT by BFA was also observed in a rounding assay of Y1 adrenal cells. The inhibition of CT cytotoxicity by BFA was dose dependent, with the ID50 value similar to the LD50 of BFA in Y1 adrenal cells. Binding and internalization of [125I]-labeled cholera toxin in Y1 adrenal cells was not affected by BFA. Unlike the BFA-sensitive cell lines such as Y1 adrenal and CHO cells, BFA at 1 micrograms/ml did not inhibit the cytotoxicity of CT in PtK1 cells, of which the Golgi structure was BFA-resistant. These results strongly suggest that a BFA sensitive Golgi is required for the protection of CT cytotoxicity by BFA. In contrast, elevation of the intracellular cAMP by forskolin, which acts directly on the plasma membrane adenylate cyclase, was not affected by BFA. These observations indicate that the intoxication of target cells by CT requires an intact Golgi region for its intracellular trafficking and/or processing. In this respect, CT shares a common intracellular pathway with ricin, Pseudomonas toxin, and modeccin, even though their structures and modes of action are very different. PMID- 8425905 TI - Induction of glucose regulated proteins during growth of a murine tumor. AB - Chronic anoxia, glucose starvation, low pH, and numerous other conditions induce the glucose-regulated system of stress proteins (GRPs), whose principal members are observed at 78, 94, and 170 kDa. These stresses may be expected to occur during growth in untreated tumors. To examine the possibility that GRPs are correspondingly induced, we have examined the protein profiles of small (< 0.1 g), intermediate (0.2-0.8 g), and large (> 1.8 g) radiation-induced fibrosarcoma (RIF) tumors grown on C3H mice. One and two-dimensional gel electrophoresis indicate that the principal GRPs at 78 and 94 are coordinately and substantially increased in large tumor masses, relative to the small, and may be partially increased in the intermediate tumors. Necrotic material removed from large tumors exhibited an identical pattern of GRP induction with no visible indication of protein degradation and also contained a significant fraction of viable cells. Western blot analysis using rabbit antisera raised against the 78 and 170 kDa GRPs also demonstrated the enhanced accumulation of these proteins in the large tumors. The antibody against the 170 kDa GRP was also capable of detecting the induction of this stress protein in large tumors by indirect immunofluorescence analysis. Northern blot studies using a probe for the GRP 78 gene also showed an increase in GRP 78 message in large tumors as well as in RIF cells exposed to anoxic stress in vitro. Two-dimensional gel electrophoresis indicated that the major heat shock proteins at 70 and 90 kDa were not increased in the larger tumors, and the amount of the 90 kDa species was reduced. Finally, the quantity of vimentin and its degradation products is significantly diminished in large tumors and in anoxic cells. This study demonstrates that RIF tumor cells undergo a glucose regulated stress response in situ during tumor growth. PMID- 8425906 TI - Structural reaction pattern of hepatocytes following exposure to hypotonicity. AB - Isolated rat hepatocytes were exposed to hypotonic media (225 mosmol/l) for 5 and 15 min and processed for a quantitative electron microscopic stereologic analysis. Within 5 min of hypotonicity, the hepatocyte volume increased by 25% and thereafter displayed a volume regulatory decrease leading to mean cellular volume, which was 16% above that of controls. Stereologic analysis of the major subcellular compartment, the cytosol, showed an identical change as the whole cell. In contrast to that, the mitochondrial compartment increased in volume by 30% within the first 5 min of exposure and returned by regulatory volume decrease back to values of the isotonic controls after 15 min of hypotonicity. In contrast, hypotonicity (220 mosmol/l)-induced stimulation of flux through mitochondrial glutaminase and the glycine cleavage enzyme complex, as assessed by 14CO2 production from [1-14C]glutamine or [1-14C]glycine in isolated perfused rat liver persisted throughout a 15-min period of hypotonic exposure. Thus hypotonicity-induced alterations of mitochondrial metabolism apparently do not parallel the time course of mitochondrial volume changes. This suggests that persistent mitochondrial swelling is not required for functional alterations, but that the latter may be triggered by the initial swelling of mitochondria. Hypotonic exposure did not alter the nuclear volume of isolated hepatocytes. Cell membrane surface nearly doubled after 5 min of hypotonic exposure, but returned within 15 min of exposure to values observed in normotonic media. This may reflect the participation of exocytosis in hepatocyte volume regulation. PMID- 8425907 TI - Regulation of matrix synthesis rates by the ionic and osmotic environment of articular chondrocytes. AB - Chondrocytes in cartilage are embedded in a matrix containing a high concentration of proteoglycans and hence of fixed negative charges. Their extracellular ionic environment is thus different from that of most cells, with extracellular Na+ being 250-350 mM and extracellular osmolality 350-450 mOsm. When chondrocytes are isolated from the matrix and incubated in standard culture medium (DMEM; osmolality 250-280 mOsm), their extracellular environment changes sharply. We incubated isolated bovine articular chondrocytes and cartilage slices in DMEM whose osmolality was altered over the range 250-450 mOsm by Na+ or sucrose addition. 35S-sulphate and 3H-proline incorporation rates were at a maximum when the extracellular osmolality was 350-400 mOsm for both freshly isolated chondrocytes and for chondrocytes in cartilage. The incorporation rate per cell of isolated chondrocytes was only 10% that of chondrocytes in situ both 4 and 24 hours after isolation. For freshly isolated chondrocytes, the rate increased 30-50% in DMEM to which NaCl or sucrose had been added to increase osmolality. In chondrocytes incubated overnight in DMEM, the rate was greatest in DMEM of normal osmolality and fell from the maximum in proportion to the change in osmolality. The effects of sucrose addition on incorporation rates were similar but not identical to those of Na+ addition. Changes in cell volume might be linked to changes in synthesis rates since the cell volume of chondrocytes (measured by Coulter-counter) increased 30-40% when the cells were removed from their in situ environment into DMEM. Synthesis rates can thus be partly regulated by changes in extracellular osmolality, which in cartilage is controlled by proteoglycan concentration. This provides a mechanism by which the chondrocytes can rapidly respond to changes in extracellular matrix composition. PMID- 8425908 TI - Kinetics of 125I-PDGF binding and down-regulation of PDGF receptor in arterial smooth muscle cells derived from patients with moyamoya disease. AB - Progressive stenosis or occlusion of bilateral internal carotid arteries by fibrocellular intimal thickening results in cerebral ischemia in moyamoya disease. We recently found that cultured smooth muscle cells (SMC) derived from arteries of patients with moyamoya disease responded poorly to serum mitogens, especially to platelet-derived growth factor (PDGF). In the present study, we investigated further the binding and processing of 125I-PDGF, as well as down regulation of the PDGF receptor in arterial SMC derived from patients with moyamoya disease. The specific binding sites of 125I-PDGF were reduced significantly at both 4 degrees C and 22 degrees C on SMC from moyamoya disease compared with those from control (4.78 vs. 11.92 x 10(4)/cell at 4 degrees C), though the apparent dissociation constant (Kd) were the same. Kinetics of 125I PDGF binding at 37 degrees C in cells from moyamoya disease showed fewer binding sites (less than 1/3 of controls) and lower degradation per cell than in those from controls, though no difference was observed in either internalization or degradation of each receptor. When SMC were exposed to lower concentrations of nonlabeled PDGF at 37 degrees C, the percentage of remaining binding sites on cells from moyamoya disease was significantly less than that from controls. This excess down-regulation of PDGF receptor in SMC from moyamoya disease may be interpreted as insufficient recycling or a decreased intracellular pool of the PDGF receptor. These results are closely correlated with the diminished proliferation responses to PDGF in SMC from moyamoya disease and provide evidence that functional alterations in vascular cells are involved in the mechanism of development of intimal thickening in moyamoya disease. PMID- 8425909 TI - Macrophage supernatants have both stimulatory and suppressive effects on mesangial cell proliferation. AB - Macrophages may modulate mesangial expansion following renal injury via secretory products. We undertook the present study to determine the effects of macrophage supernatants on mesangial cell proliferation. Macrophage supernatants collected in serum-free media after 24 hours caused significantly enhanced mesangial cell proliferation in long-term culture at concentrations up to 50% but caused suppression at higher concentrations (control, 122,000 +/- 14,000 cells/well; 50% supernatant, 188,000 +/- 15,100 cells/well, P < 0.02 compared to control, n = 4; 80% supernatant, 52,000 +/- 3,500 cells/well, P < 0.01 compared to control, n = 4). In short-term culture [3H]thymidine incorporation, a measure of DNA synthesis, was significantly enhanced compared to control at supernatant concentrations up to 30% (30% supernatant, 4,120 +/- 310 cpm/well; control, 3,210 +/- 97 cpm/well, P < 0.5, n = 4), but uptake was reduced at high concentration (80% supernatant, 2,900 +/- 74 cpm/well; control, 3,210 +/- 97 cpm/well, P < 0.05, n = 4). When macrophage supernatants were collected after 48 hours incubation and incubated with mesangial cells, mesangial cell thymidine uptake was significantly suppressed compared to control (48-hour supernatant, 4,060 +/- 260 cpm/well; control, 5,890 +/- 270 cpm/well, P < 0.01, n = 4) and compared to 24-hour supernatants, which enhanced uptake (24-hour supernatant, 8,080 +/- 340 cpm/well; control, 5,890 +/- 270 cpm/well, P < 0.01, n = 4). Our results suggest that macrophage supernatants can directly enhance mesangial cell proliferation in vitro in both short-term and long-term culture, though this effect is lost at high concentrations of supernatant. These data lend support to the potential role of the macrophage in mediating mesangial expansion following renal injury. PMID- 8425910 TI - Timing of protooncogene expression varies in toxin-induced liver regeneration. AB - Hepatic expression of the protooncogenes c-fos and c-myc occurs within 2 h after partial hepatectomy, and these immediate early genes are thought to prime the hepatocytes for subsequent proliferation. To examine whether such gene activation occurred in the setting of hepatocyte proliferation after toxic liver injury, protooncogene expression was examined during the regenerative response following liver injury from carbon tetrachloride (CCl4) or galactosamine (GalN). The pattern of protooncogene expression after CCl4 mirrored that seen after partial hepatectomy, with rises in c-fos and c-myc mRNA content within 2 h, and then a rapid return to baseline levels. In contrast, early c-fos and c-myc expression did not occur after GalN injury. Instead GalN-induced regeneration led to a delayed, and prolonged c-fos and c-myc activation which peaked 24-48 h after injury. Increases in c-jun, jun-B, and jun-D mRNA levels also occurred in both models at times similar to the rises of c-fos and c-myc expression. Although the timing of DNA synthesis was identical after GalN or CCl4 treatment, the proliferative response after GalN injury was significantly less than that of CCl4, and marked by the histologic appearance of oval cells. The coadministration of 2-acetylaminofluorene, an inhibitor of differentiated hepatocyte proliferation, together with CCl4 altered the usual pattern of post-CCl4 protooncogene expression to one resembling that seen after GalN injury. Thus, the timing of protooncogene expression during liver regeneration may vary considerably. These variations may influence the nature of the proliferative response in terms of which cell type(s) proliferates, and the amount of regeneration that ensues. PMID- 8425911 TI - Effect of low extracellular Ca2+ on growth, spreading area, cytoplasmic Ca2+ concentration, and intracellular pH in normal and transformed human fibroblasts. AB - The transformation of certain cells reduces the requirement of extracellular Ca2+ for growth. The SV-40 transformed human lung fibroblasts, WI-38 VA13, require less Ca2+ than normal WI-38 cells. Spreading area of the normal cells decreases when cultured in 10 microM Ca2+ medium. Intracellular calcium concentration ([Ca2+]i) of the normal and transformed cells cultured in 10 microM and 2 mM Ca2+ media was measured by the fluorescence microscope technique using fura-2 as a probe. The [Ca2+]i is measured in the resting state and during mobilization by serum or bradykinin stimulation. The lowering of extracellular calcium concentration results in a decrease in the resting state [Ca2+]i of both normal and transformed cells. Although the total decrease in [Ca2+]i is the same for both cells, the rate of decrease is much faster in normal cells than in transformed cells. Low extracellular Ca2+ reduces the number of cells responsive to the serum or bradykinin stimulation and decreases the peak [Ca2+]i value in both cells. In addition, we investigated, using BCECF as a fluorescent probe, the intracellular pH (pHi) of normal and transformed cells maintained at low and normal Ca2+. The low Ca2+ condition makes pHi acidic in normal cells but not in transformed cells. The acidification of the normal cell is accompanied by a decrease in the spreading area of the cells. The decrease of the cell attachment, followed by the reduced spreading area, induces the acidic pHi. These results suggest that the reduced Ca2+ requirement of transformed cells for growth is related to the mechanism of pHi regulation rather than Ca2+ homeostasis and, possibly, to the anchorage-independent growth, which is a unique feature of transformed cells. PMID- 8425912 TI - Osteoblastic gene expression during adipogenesis in hematopoietic supporting murine bone marrow stromal cells. AB - A growing body of data suggests that the bone marrow stroma contains a population of pluripotent cells capable of differentiating into adipocytes, osteoblasts, and lymphohematopoietic supporting cells. In this work, the murine stromal cell lines BMS2 and +/+ 2.4 have been examined as preadipocytes and adipocytes for evidence of osteoblastic gene expression. Adipocyte differentiation has been quantitated using fluorescence activated cell sorting. Within 7-10 days of adipocyte induction by treatment with glucocorticoids, indomethacin, and methylisobutylxanthine, between 40% to 50% of the cells contain lipid vacuoles and exhibit a characteristic adipocyte morphology. Based on immunocytochemistry, both the adipocytes and preadipocytes express a number of osteoblastic markers; these include alkaline phosphatase, osteopontin, collagen (I, III), bone sialoprotein II, and fibronectin. Based on biochemical assays, the level of alkaline phosphatase expression is not significantly different between preadipocyte and adipocyte cells. However, unlike rat cell lines, dexamethasone exposure causes a dose-dependent decrease in enzyme activity. The steady-state mRNA levels of the osteoblast associated genes varies during the process of adiopogenesis. The relative level of collagen I and collagen III mRNA is lower in adipocyte-induced cells when compared to the uninduced controls. Osteocalcin mRNA is detected in preadipocytes but absent in adipocytes. These data indicate that osteoblastic gene expression is detected in cells capable of undergoing adipocyte differentiation, consistent with the hypothesis that these cell lineages are interrelated. PMID- 8425913 TI - Increase in portal flow induces c-myc expression in isolated perfused rat liver. AB - We examined expression of the c-myc oncogene in isolated perfused livers to elucidate the mechanisms involved in triggering the proliferation of hepatocytes after partial hepatectomy (PH). During perfusion with a 1:1 mixture of Dulbecco's modified Eagle's medium and the oxygen transport fluid FC-43, rat livers were two thirds resected (PH), and further perfused for 1 1/2 hours at the physiological portal flow throughout the perfusion. Expression of c-myc in the perfused livers with PH(+) was ten times higher than in those with PH(-). Furthermore, expression of c-myc in the PH(-) livers perfused with a threefold volume of the physiological portal flow was 5-10 times higher than that in the livers perfused with the physiological portal flow. The perfusates that passed through the livers did not induce DNA synthesis of primary cultured hepatocytes. These results suggest that an increase in the portal flow volume may act as a trigger for hepatocyte proliferation after PH. PMID- 8425914 TI - Cell cycle-dependent gene expression in V point-arrested BALB/c-3T3 cells. AB - Density-arrested BALB/c-3T3 cells stimulated to proliferate in an amino acid deficient medium arrest in mid-G1 at a point termed the V point. Cells released from V point arrest require 6 hr to traverse late G1 and enter S phase. As data presented here show that mRNA synthesis is needed for 2-3 hr after release of cells from the V point, after which inhibition of mRNA synthesis does not prevent entry into S phase, we used this mid-G1 arrest protocol to analyze gene expression in late G1. We found that although stimulation of cells in amino acid deficient medium did not inhibit the induction of genes expressed in early G1, genes normally expressed in late G1 were expressed only after release from the V point. The expression of late G1 genes in cells released from the V point was temporally similar, in respect to G1 location, as was seen in stimulation of quiescent G0 cells. As this protocol effectively divides gene expression into early (pre-V point) and late (post-V point) categories, it should be useful in studies of growth factor-modulated events that regulate traverse of late G1 and commitment to DNA synthesis. In addition, we used c-myb antisense oligonucleotides to show that c-myb expression, which occurs in late G1, is required for BALB/c-3T3 fibroblasts to traverse late G1 and initiate DNA synthesis. PMID- 8425915 TI - Coordinate regulation of mRNAs from multiple calmodulin genes during myoblast differentiation in vitro. AB - Multiple genes encoding identical calmodulin molecules have been found in all mammalian species so far examined, but little is known regarding the factors involved in regulating the expression of this gene family. We have investigated the possibility of differential regulation under conditions of cell cycle withdrawal and differentiation in the nonfusing BC3H1 myoblast. Transcripts from the three genes are expressed in myoblasts and myocytes and each of the mRNA species decreases during BC3H1 differentiation. Calmodulin protein levels also decrease, although with distinct kinetics with respect to the mRNAs. Previous studies indicated that a decrease in transcription is involved (Epstein et al., Molecular Endocrinology 3:193-202, 1989). In this study, an increase in stability for each of the mRNA species is also shown to contribute to overall mRNA levels. The calmodulin mRNAs are also found to decrease under conditions of cell cycle withdrawal when differentiation is blocked. This demonstrates that the expression of mRNA from all three genes is directly coupled with the proliferation state but only indirectly with the differentiation state. Consistent with this, calmodulin expression decreases in serum deprived fibroblasts as they exit the cell cycle. PMID- 8425916 TI - Modulation of the epidermal growth factor receptor by basic fibroblast growth factor. AB - Treatment of Swiss 3T3 fibroblasts with basic fibroblast growth factor (bFGF) lead to a rapid reduction in epidermal growth factor (EGF) binding and a slower inhibition of EGF receptor autophosphorylation. The reduction in binding was due to a complete loss of the highest affinity EGF binding sites and a reduction in the lower affinity binding sites. Neither the inhibition of EGF binding nor the inhibition of EGF receptor autophosphorylation required protein kinase C. Treatment of cells with bFGF stimulated the phosphorylation of the EGF receptor, which persisted for several hours. The inhibition of EGF receptor autophosphorylation by bFGF was reduced in the presence of cycloheximide. However, cycloheximide had no effect on the reduction of EGF binding by bFGF. In contrast to these results with Swiss 3T3 fibroblasts, treatment of PC12 cells with bFGF lead to a reduction in EGF binding but no inhibition of EGF receptor autophosphorylation. Thus inhibition of EGF receptor autophosphorylation and inhibition of EGF binding can be uncoupled. PMID- 8425917 TI - Sex-dependent effects of 17-beta-estradiol on chondrocyte differentiation in culture. AB - This study examined the effects of 17-beta-estradiol (E2) on chondrocyte differentiation in vitro. Cells derived from male or female rat costochondral growth zone and resting zone cartilage were used to determine whether the effects of E2 were dependent on the stage of chondrocyte maturation and whether they were sex-specific. [3H]-Thymidine incorporation, cell number, alkaline phosphatase specific activity, and percent collagen production were used as indicators of differentiation. Alkaline phosphatase specific activity in matrix vesicles and plasma membranes isolated from female chondrocyte cultures was measured to determine which membrane fraction was targeted by the hormone. Specificity of the E2 effects was assessed using 17-alpha-estradiol. The role of fetal bovine serum and phenol red in the culture medium was also addressed. The results demonstrated that E2 decreases cell number and [3H]-thymidine incorporation in female chondrocytes, indicating that it promotes differentiation of these cells. Alkaline phosphatase specific activity is stimulated in both growth zone and resting zone cells, but the effect is greater in the less mature resting zone chondrocytes. The increase in enzyme activity is targeted to the matrix vesicles in both cell types, but the fold increase is greater in the growth zone cells. In male chondrocytes, there was a decrease in [3H]-thymidine incorporation at high E2 concentrations in resting zone cells at the earliest time point examined (12 hours) and a slight stimulation in alkaline phosphatase activity in growth zone cells at 24 hours. Cells cultured in serum-free medium exhibited a dose-dependent inhibition in alkaline phosphatase activity when cultured with E2, even in the presence of phenol red. E2-dependent stimulation of enzyme activity is seen only in the presence of serum, suggesting that serum factors are also necessary. E2 increased percent collagen production in female cells only; the magnitude of the effect was greatest in the resting zone chondrocyte cultures. The results of this study indicate that the effects of E2 are dependent on time of exposure, presence of serum, and the sex and state of maturation of the chondrocytes. E2-dependent stimulation of alkaline phosphatase specific activity is targeted to matrix vesicles. PMID- 8425918 TI - Angiotensin II-induced vascular smooth muscle cell hypertrophy: PDGF A-chain mediates the increase in cell size. AB - We report here that angiotensin II-mediated hypertrophy of vascular smooth muscle cells (VSMC) exhibits PDGF A-chain-dependent and -independent pathways. Secretion of PDGF A-chain is required for the increase in cell size, but not for the increase in protein synthesis. Angiotensin II stimulates a hypertrophic growth response in VSMC characterized by increases in cell size and protein synthesis, but not cell number. Because angiotensin II-stimulated VSMC hypertrophy has been associated with increased PDGF A-chain expression, we studied its role in the hypertrophic response by inhibiting PDGF A-chain expression with hydrocortisone or anti-PDGF antibody. Hydrocortisone (1 microM for 48 h) inhibited basal protein synthesis by 47%, but angiotensin II-stimulated protein synthesis was enhanced (111% increase after hydrocortisone treatment vs. 25% increase in control). In contrast, hypertrophy, as measured by cell size, was completely inhibited. Although hydrocortisone had no effect on early growth signals stimulated by angiotensin II (e.g., activation of protein kinase C, stimulation of Na+/H+ exchange, and c-fos and c-myc expression), it significantly decreased angiotensin II-stimulated secretion of PDGF-like material into the medium from 0.4 to 0.1 ng/ml/24 h (p < 0.01). However, the time course for PDGF secretion (maximal at 16 24 h) was significantly slower than the time course for angiotensin II-stimulated protein synthesis (maximal at 4-12 h). To block the action of PDGF A-chain selectively, VSMC were treated with anti-PDGF A-chain antibody. The antibody completely inhibited the angiotensin II-stimulated increase in cell size, but it had no significant effect on protein synthesis at early times (< 8 h). These findings demonstrate two pathways involved in angiotensin II-stimulated VSMC hypertrophy: an increase in cell size dependent on PDGF A-chain and an increase in protein synthesis independent of PDGF A-chain. PMID- 8425919 TI - Establishment of a hepatocytic epithelial cell line from the murine fetal liver capable of promoting hemopoietic cell proliferation. AB - Although the fetal liver has been thought to be the main hemopoietic organ in the embryonal period, whether or not hepatocytes play a major role in hemopoiesis remains obscure. We have established an epithelial cell line from the murine fetal liver, which can support hemopoiesis in vitro. The proliferation of the epithelial cells was promoted synergistically by both epidermal growth factor (EGF) and insulin. The cells were identified as epithelial cells by the presence of desmosomes and tight junctions. Cytoplasmic organelles including small mitochondria and dilated Golgi apparati as well as intercellular canalicular structures similar to bile canaliculus also helped in confirming the hepatic origin of the cell line (designated as FHC). The cells in the primary culture were positive for both alpha-fetoprotein and albumin, indicating the hepatocytic nature of the cell line. Cloned FHC cells were demonstrated to have the ability to maintain hemopoietic progenitors in fetal liver and adult bone marrow in the coculture, and among them, FHC-4D2 clone displayed the greatest activity. Hemopoiesis-supporting function could also be seen even when bone marrow cells were separated from FHC-4D2 cells by nitrocellulose membrane. Column chromatography revealed three distinct peaks of hemopoietic activities with different molecular sizes in the supernatant of FHC-4D2. Neutralization test with antibodies and proliferative response to interleukin-3 (IL-3)/granulocyte macrophage colony stimulating factor (GM-CSF)-responding IC2 cells demonstrated that the hemopoietic activities were attributed to GM-CSF and macrophage colony stimulating factor (M-CSF). Transcripts of GM-CSF and M-CSF were readily detectable in Northern blot analysis, whereas no messages for IL-3, IL-6, CSF for granulocytes (G-CSF) or erythropoietin (EPO) were identified. Therefore, this is the first report on the fetal hepatocyte cell line capable of supporting hemopoiesis. PMID- 8425920 TI - Orthovanadate both mimics and antagonizes the transforming growth factor beta action on normal rat kidney cells. AB - Normal rat kidney [NRK] cells grown in the presence of epidermal growth factor (EGF) or platelet-derived growth factor (PDGF) have a normal phenotype and undergo density-dependent growth inhibition, whereas in the presence of multiple growth factors, density arrest is lost and the cells become phenotypically transformed. We studied the influence of the protein tyrosine phosphatase (PTPase) inhibitor sodium orthovanadate on the mitogenic stimulation of NRK cells by growth factors and on transformation-linked properties as loss of density dependent growth inhibition and anchorage-independent growth. The fraction of cells in serum-deprived monolayer cultures that is induced to proliferate upon mitogenic stimulation by EGF or PDGF is only slightly enhanced upon addition of low concentrations (25-50 microM) of vanadate. Addition of vanadate per se induces proliferation of only a very limited amount of cells, but results in a shift of the dose-response curves for other growth factors to lower concentrations. Vanadate added in combination with EGF or PDGF is able to mimic the effect of transforming growth factor beta (TGF beta) in inducing phenotypic transformation. In monolayer cultures density-dependent growth inhibition is lost and anchorage-independent proliferation is observed on dishes coated with poly(2 hydroxy-ethyl methacrylate) (polyHEMA). The extent of these changes is similar to that induced by TGF beta. However, the morphology of the obtained colonies in polyHEMA-coated dishes is quite different. Cells transformed by TGF beta in the presence of EGF form rather amorphous colonies, whereas in the presence of orthovanadate colonies are formed that tend to fall apart in loose cells. The effect of vanadate on cell transformation is dependent on the growth factor conditions in a bimodal way. When a suboptimal dose of growth factor(s) is used, 25 microM vanadate is very effective in preventing density-induced growth inhibition and stimulating anchorage-independent proliferation. However, the same concentration of vanadate is inhibitory when cells are maximally stimulated and antagonizes the transforming effect of TGF beta added in combination with other growth factors. It is hypothesized that vanadate acts on a set of different protein tyrosine phosphatases. Some of these are positive and others negative regulators of growth. PMID- 8425921 TI - Effect of amino acid analogs on the development of thermotolerance and on thermotolerant cells. AB - Exposure of HA-1 Chinese hamster fibroblasts to amino acid analogs has been shown to have a heat-sensitizing effect as well as inducing the heat shock response (Li and Laszlo, 1985a). In this study, we have examined the effect of amino acid analogs on the development of thermotolerance after a brief heat shock or exposure to sodium arsenite and the effect of amino acid analogs on cells that are already thermotolerant. Exposure of HA-1 cells to amino acid analogs inhibited the development of thermotolerance following a mild heat shock or treatment with sodium arsenite. However, cells that were already thermotolerant were resistant to the sensitizing action of amino acid analogs. The refractoriness of thermotolerant cells to amino acid analog treatment developed in parallel with thermotolerance. The uptake of the arginine analog, canavanine, and its incorporation into proteins was not altered in the thermotolerant cells. Furthermore, another biological consequence of exposure to amino acid analogs, sensitization to ionizing radiation, also was not altered in the thermotolerant cells. The inhibition of the development of thermotolerance by amino acid analogs and the refractoriness of thermotolerant cells to the heat-sensitizing action of amino acid analogs lend further support the role of heat-shock proteins in the phenomenon of thermotolerance. PMID- 8425922 TI - Cytokine-induced expression of mRNAs for chemotactic factors in human synovial cells and fibroblasts. AB - In response to interleukin 1 or tumor necrosis factor, human synovial cells and fibroblasts expressed several genes encoding known chemotactic factors or related proteins. Transcripts for interleukin 8 (IL-8), gro/MGSA, pAT 464, IP-10, pAT 744 and Monocyte Chemotactic and Activating Factor (MCAF) accumulated rapidly in IL-1 and TNF-treated cells. The inhibition of protein synthesis led to the superinduction of IL-8 and gro/MGSA mRNAs in IL-1, but not in TNF-treated cells. Thus, IL-1 and TNF are likely to regulate the expression of these mRNAs by different mechanisms. Important cell-specific differences in mRNA accumulation characterized the expression of chemotactic factor genes. Moreover, only a subset of the same genes was activated in quiescent cells stimulated by serum. Therefore, genes encoding closely related proteins each had a distinct pattern of expression. continuous stimulation of fibroblasts and synovial cells with IL-1 resulted in high and prolonged expression of IL-8 and gro/MGSA mRNAs. These results extend the list of chemotactic factor genes expressed by mesenchymal cells in vitro and suggest a pivotal role for these cells in processes such as chronic inflammation. PMID- 8425923 TI - Simulated quantitative and qualitative isotachophoretic indices of 73 amino acids and peptides in the pH range 6.4-10. AB - Qualitative and quantitative isotachophoretic indices of 73 amino acids, dipeptides and tripeptides were simulated under 24 leading electrolyte conditions covering the pH range 6.4-10. The RE values and time-based zone lengths are tabulated together with the absolute mobility (m0) and pKa values used. The leading electrolyte used was 10 mM HCl and the pH buffers were imidazole, tris(hydroxymethylamino)methane, 2-amino-2-methyl-1,3-propanediol and ethanolamine. The simulated indices will be useful in the assessment of the separability and determination of the listed and related compounds. PMID- 8425924 TI - High-performance liquid chromatographic determination of urinary 2,5-hexanedione. PMID- 8425925 TI - Patterns of performance on the verbal and performance subtests of the Wechsler Adult Intelligence Scale-Revised: some Australian data. AB - The pattern of performance of 345 vocational guidance clients on the WAIS-R was examined in relation to previous studies of client populations. An exploratory principal axis factor analysis was conducted, and two factors that accounted for 89% of the total variance were selected. These comprised a general factor on which the Verbal subtests loaded most highly and a smaller Performance subtest factor. A hypothesis-testing approach was undertaken as a second step, using multiple group analysis. Two factors (which correlated .58) accounted for 65% of the total variance. The first factor (51%) was also a general factor on which all subtests, but moreso the Verbal tests, loaded highly. The second factor was dominated by Performance subtests. Implications for special client samples and intellectual assessment are discussed. PMID- 8425926 TI - Reductions in turnover, accidents, and absenteeism: the contribution of a pre employment screening inventory. AB - This study examined the rates of turnover, work-related accidents, and unauthorized absence among two contrasted groups of employees in a mid-sized manufacturing environment. For the calendar year 1989, comparative rates on each variable were obtained for a group hired prior to the inclusion of a pre employment inventory in the Company's selection process and a group hired subsequently. Results indicate that rates of turnover, accidents, and unauthorized absence were all significantly lower in the group hired subsequent to inclusion of the pre-employment screening inventory. Some possible design artifacts and cost-benefit implications are discussed. PMID- 8425927 TI - Judged levels of success upon discharge from Department of Veterans Affairs domiciliaries. AB - To help define the goals for domiciliary patients and programs, staff who were working in Department of Veterans Affairs Domiciliary programs across the country were sampled. Each employee was asked to rank order the success of different outcomes for veterans who were being treated in the domiciliary. The rankings are compared for uniformity and displayed as perceived successful and unsuccessful outcomes by staff who work with domiciliary patients and staff who do not work with them. PMID- 8425928 TI - Computer content analysis of the Schreber case. AB - The text of Schreber's Memoirs of My Nervous Illness was analyzed by computer at the level of the individual word. These words then were grouped into 17 rational categories, and the categories were checked for reliability. The contents of Schreber's work then were compared with three other documents. In general, the Memoirs showed much greater delusional content than the other documents. Interestingly, sexual matters did not appear to be Schreber's principal problem at this atomistic level. PMID- 8425929 TI - Development and validation of a Multidimensional Sense of Humor Scale. AB - Personal sense of humor has been recognized as a coping mechanism. Sense of humor as a construct, however, is multidimensional. Existing efforts to assess overall sense of humor are, for a number of reasons, deficient. The present study reports the development and factor analysis of a new Multidimensional Sense of Humor Scale that may be useful in a variety of research and clinical applications. PMID- 8425930 TI - Comparison of MMPI-2 scores of foreign Chinese and Caucasian-American students. AB - MMPI-2 scores of foreign Chinese students (n = 25) were compared to those of a matched sample of Caucasian students (n = 21) and to normative data on American college students. Although responses for all groups were within normal limits, Chinese men appeared more socially introverted than Caucasian men. Relative to Caucasian women, Chinese women were more defensive, depressed, unaware of somatic and psychosocial problems, and gender astereotypic in interests. These tentative findings are discussed in terms of ethnicity and adjustment. PMID- 8425931 TI - Validation of the MCMI-I Borderline Personality Disorder Scale with a well defined criterion sample. AB - This study examined the MCMI-I BPD scale's accuracy in assigning borderline personality disorder as a primary diagnosis. Clinicians with particular expertise in Axis II pathology diagnosed borderline patients in two groups in this sample. Results indicate that the MCMI-I BPD scale has very limited utility as a screening instrument for individual diagnoses when a well-defined borderline sample is-used as a criterion. This study further suggests that at present only clinicians with particular expertise in the diagnosis of personality disorders can assess the degree and type of Axis II pathology present in a given patient. Because most structured interview and self-report validation studies have not included well-defined criterion groups, diagnostic validity of particular measures generally has not been established beyond the level of concordance. PMID- 8425932 TI - Emotional experiences during the three trimesters of pregnancy. AB - This study examined the emotional changes that occur during the trimesters of pregnancy. Two hundred eighty-two women were asked, one day after giving birth, to indicate at what frequency they had experienced various symptoms during each trimester of pregnancy and to fill out the Repression-Sensitization scale (Byrne, Barry, & Nelson, 1963). Results showed that while women's feelings during the first trimester are characterized by symptoms related to physiological changes (e.g., nausea, vomiting, dizziness), during the last trimester anxiety and emotional distress become the most significant symptoms. The level at which these symptoms were experienced was affected by the subject's socioeconomic level, number of previous births (primaparae or multiparae), and her personality type (repressor or sensitizer). PMID- 8425933 TI - Complex partial epileptic signs as a continuum from normals to epileptics: normative data and clinical populations. AB - Over a 10-year period, a total of 447 men and 624 women between 18 and 61 years of age were administered an inventory whose items describe experiences that are similar to those evoked by electrical stimulation of the temporal lobes. Empirically determined factors contained experiences of sensory enhancement, affective-dissociation, ego alien intrusions, and literary emphasis. Using this population as a reference, T scores for these clusters were calculated for special normal populations (poets, drama students, false pregnancies) and for clinical groups (post-traumatic stress, anxiety-depersonalization, exotic dissociations, and complex partial epilepsy). Whereas only mild elevations (50 < T < 65) in indicators of temporal lobe signs and symptoms were noted in the special groups, moderate (65 < T < 75) and severe (T > 79) elevations were noted in the clinical populations. PMID- 8425934 TI - Malingering on neuropsychological memory tests: potential objective indicators. AB - We evaluated five potential indicators of malingering on the Rey Memory Test (RMT), Hebb's Recurring Digits (HRD), the Wechsler Memory Scale-Revised (WMS-R), the Complex Figure Test (CFT), and the Auditory Verbal Learning Test (AVLT). Fifty-seven subjects were assigned randomly to either a control group or a simulated malingering group. Two indicators-discriminant functions derived from the WMS-R and from the CFT/AVLT-achieved classification accuracy of 88% and 86%, respectively, without misidentifying controls as malingerers. Among the remaining indicators, there were problems with the recommended RMT cut-off, but it and the HRD show some promise. Subjects who simulated malingering did so by suppressing performance on tasks that involve recall as well as recognition memory and are relatively easy, but not obviously so. PMID- 8425935 TI - Wisconsin Card Sorting Test performance in healthy, older adults: relationship to age, sex, education, and IQ. AB - We obtained Wisconsin Card Sorting Test (WCST) scores on 91 (35 male and 56 female) healthy, well-educated individuals aged 45 to 83. Women scored better than men on six WCST scores (categories, errors, perseverative responses, % perseverative errors, % conceptual level responses, and trials to first category), while subjects with more than 16 years of education outscored those limited to high school education on four measures (perseverative responses, errors, % perseverative errors, % conceptual level responses). Individuals older than 70 years of age scored less well than younger subjects on only two indices (errors, % conceptual level responses). Full Scale IQ was not related to WCST scores. Our findings suggest that WCST scores should be interpreted within the context of patient gender, education, and age. PMID- 8425936 TI - Factor structure and scoring of the SKT test battery. AB - The SKT (Syndrom Kurztest) has been used in the assessment of treatment responses in numerous clinical trials for treatment of dementia in German-speaking Europe. Data from 265 patients with mild to moderate Alzheimer's disease in a study conducted in the U.S. were analyzed to evaluate factor structure, common and specific subtest content, reliability, and concurrent validity. Results confirm the presence of two primary factors of memory and attention. Test-retest reliability of the factor scores was estimated to be .75 and .93. Test-retest reliability of the composite SKT total score was .90. The correlations between the SKT memory and attention factor scores and the MMSE and ADAS measurements of dementia also support validity with regard to the broader construct of cognitive dysfunction. PMID- 8425937 TI - Concurrent validity of the Cognitive Levels Test with measures of achievement for a sample of referred children and youth. AB - The present investigation compared the Cognitive Levels Test (CLT) with the Woodcock Reading Mastery Test (WRMT) and Wide Range Achievement Test-Revised (WRAT-R) for 55 children and youth who were referred for remedial/special education services. A comparison of the correlations for the criterion measures for the WRMT and WRAT-R showed a consistent significant positive relationship with the CLT. A repeated measures analysis of variance that compared standard scores of the CLT with those of the WRMT and WRAT-R indicated that significant differences existed between the test scores. Implications with respect to the concurrent validity of the CLT are discussed. PMID- 8425938 TI - The Neurobehavioral Cognitive Status Examination: psychometric properties in use with psychiatric inpatients. AB - The present investigation sought to enhance clinical utility of the Neurobehavioral Cognitive Status Examination (NCSE; Northern California Neurobehavioral Group, Inc.) by providing reference scores for an inpatient psychiatric sample and assessing construct validity. A total of 866 patients (aged 15-92 years) received an NCSE 2 to 4 days after admission. Examination of means, standard deviations, z scores, and percent who passed each screening item revealed consistently poorer performance for psychiatric patients relative to the original normative sample. Pearson product-moment correlations between age and each NCSE subtest similarly yielded significant negative correlations, particularly on tests predicted to be differentially sensitive to aging. Intercorrelations between subtests, however, failed to yield expected patterns of performance. We conclude that the NCSE provides a moderately valid screening instrument for cognitive impairment. PMID- 8425939 TI - The development of verbal and nonverbal factorially derived memory measures. AB - Sets of verbal and nonverbal memory tests were subjected to a factor analysis, and the factor composite scores were used to discriminate between 57 brain damaged and 34 non-brain-damaged subjects. The derived factors clearly represented verbal and nonverbal factors. The brain-damaged group performed significantly less well on both the verbal and nonverbal factor composite scores. It is suggested that a battery of verbal and nonverbal scales be used to discriminate left and right lateralized brain-damaged patient groups. PMID- 8425940 TI - Clinical presentation of the lead-poisoned child on mental ability tests. AB - Current literature on effects of childhood lead poisoning and high blood lead level on psychological test performance was reviewed. A pathognomic configuration of mental ability test scores is described. A sample of 18 moderately to severely lead poisoned children was found to have a significantly higher frequency of this score configuration than either a comparison sample or the Black subsample of the original WISC-R standardization group. Implications for clinical practice and research are discussed. PMID- 8425941 TI - Distribution of [3H]quinuclidinyl benzilate, [3H]nicotine, and [125I]alpha bungarotoxin binding sites in the nucleus tractus solitarii of the cat. AB - The distribution of muscarinic and nicotinic cholinergic binding sites in the cat nucleus tractus solitarii was studied by the technique of in vitro autoradiography. Using the antagonist [3H]quinuclidinyl benzilate, muscarinic binding sites were differentially located in subdivisions of the nucleus tractus solitarii. The majority of muscarinic binding sites were located predominantly in the caudal half of the nucleus, reaching their greatest amounts at the mid levels of the nucleus tractus solitarii. The medial, dorsolateral, intermediate, and interstitial subdivisions contained the highest densities of quinuclidinyl benzilate binding sites. Nicotinic cholinergic binding sites, using [3H]nicotine and [125I]alpha-bungarotoxin, had unique patterns of distribution. With [3H]nicotine the majority of binding sites were located in rostral levels of the nucleus with very few binding sites present in the caudal half. In contrast, [125I]alpha-bungarotoxin binding sites were present mainly in subdivisions located in the caudal half of the nucleus, i.e., commissural, ventrolateral, dorsolateral, medial, and intermediate subdivisions, and dropped off precipitously at more rostral levels. The differential distribution of [3H]nicotine and [125I]alpha-bungarotoxin suggests the two ligands may be labeling different types of nicotinic binding sites in the nucleus tractus solitarii. The unique distribution of muscarinic and nicotinic cholinergic binding sites in the various subdivisions of the nucleus solitarii suggests that muscarinic and nicotine mechanisms may play an active role in the regulation of the diverse autonomic functions at the level of the nucleus tractus solitarii. PMID- 8425942 TI - Retinal projection to the olfactory tubercle and basal telencephalon in primates. AB - The retinal projection to the basal telencephalon was studied in eight species of primates from the suborders Strepsirhini and Haplorhini, including one anthropoid primate, the gibbon. Animals received an intraocular injection of tritiated amino acids and the distribution of retinal fibers and terminals was demonstrated by autoradiographic techniques in horizontal and coronal sections. In all species a discrete group of labeled retinal fibers is observed to branch off from the dorsolateral aspect of the optic tract at the level of the suprachiasmatic nucleus. These fibers, destined to the basal telencephalon, are topographically distinct from the retinal fibers which innervate the suprachiasmatic nucleus and medial hypothalamic regions. The fibers of the retinotelencephalic tract course dorsally above the supraoptic nucleus through the lateral hypothalamic area and then proceed further rostrally and laterally below the diagonal band of Broca towards the olfactory tubercle. Within the olfactory tubercle, terminal distribution of label is observed in the mediocaudal region along the granular cell layer II. In the macaque this cellular layer shows a characteristic thickening in the region of retinal terminals which is evident in both coronal and horizontal section. In some species this labeled region is seen within the superficial bulge of the tubercle on the ventral aspect of basal telencephalon. In all primates the retinal projection to olfactory tubercle is bilateral. In prosimians label is predominantly contralateral to the injected eye, in New World monkeys label is equally distributed on both sides of the brain and in Old World monkeys label is mainly found ipsilaterally. Retinal fibers were also seen in the periamygdaloid region but never extended as far as piriform cortex. These results, in addition to previous studies in other mammalian orders, confirm that the basal telencephalon, and in particular the olfactory tubercle, constitutes a region of visual and olfactory convergence. This sensory integration may be related to photic and chemosensory modulation of reproductive physiology and behavior. PMID- 8425943 TI - Specificity in the efferent projections of the nucleus accumbens in the rat: comparison of the rostral pole projection patterns with those of the core and shell. AB - The efferent connections of the rostral pole of the rat accumbens, where distinct core and shell subterritories can not be identified, were examined with the aid of the anterogradely transported plant lectin, Phaseolus vulgaris-leucoagglutinin (PHA-L), for comparison with the previously reported projection patterns of the accumbal core and shell. Injection sites and transported PHA-L were evaluated with the aid of reference to adjacent sections processed to display substance P or calbindin 28 kD immunoreactivities, i.e., markers that demonstrate the core and shell. Lateral parts of the rostral pole gave rise to a "core-like" projection system that involved the rostroventral globus pallidus, subcommissural ventral pallidum, entopeduncular nucleus and an adjacent part of the lateral hypothalamus, lateral ventral tegmental area, dorsal pars compacta, and structures in the lateral mesencephalic tegmentum and central grey. The medial part of the rostral pole gave rise to a "shell-like" innervation of the subcommissural ventral pallidum, lateral preoptic region, lateral hypothalamus, ventral tegmental area, dorsalmost pars compacta, retrorubral field, lateral midbrain tegmentum, and central grey. In contrast to the large numbers of axon varicosities observed through the entire length of lateral hypothalamus following shell injections, dense accumulations of axon collaterals and varicosities in hypothalamus were limited to the levels of origin of the stria medullaris bundle and entopeduncular nucleus and to the posterlateral region following medial injections. The medial part of the rostral pole contributed some projections to preoptic and sublenticular regions, but not to the bed nucleus of the stria terminalis. Noteworthy concentrations of calbindin immunoreactive cells observed in the lateral rostral pole correlate with the origin of the "basal ganglia-like" projection system, provoking the speculation that ventral striatal calbindin immunoreactive cells contribute principally to basal ganglia-like projections while cells lacking calbindin immunoreactivity contribute to the innervation of hypothalamus and midbrain tegmentum. PMID- 8425944 TI - Detection of the 5-HT1A receptor and 5-HT1A receptor mRNA in the rat bowel and pancreas: comparison with 5-HT1P receptors. AB - We tested the hypothesis that the rat bowel and pancreas contain 5-HT1A receptors. 3H-8-hydroxy-2-(di-n-propylamino)tetralin (3H-8-OH-DPAT) was used as a radioligand. Binding of 3H-8-OH-DPAT to membranes derived from the myenteric plexus and the pancreas was investigated by rapid filtration. Alternatively, radioautography was employed to locate 3H-8-OH-DPAT binding sites in frozen sections of unfixed bowel or pancreas. An excess of 5-HT (10 microM) was used to define nonspecific binding. Saturable, high affinity binding of 3H-8-OH-DPAT to enteric (Kd = 2.8 +/- 1.1 nM; Bmax = 83.8 +/- 4.3 fmol/mg protein) and pancreatic (Kd = 6.6 +/- 1.3 nM; Bmax = 44 +/- 2.2 fmol/mg protein) membranes was found. The binding of 3H-8-OH-DPAT to enteric and pancreatic membranes was inhibited by 8-OH DPAT, NAN-190, and spiperone. In contrast, the binding of 3H-8-OH-DPAT to enteric and pancreatic membranes was not inhibited by 5-carboxyamidotryptamine, or by a variety of compounds known to bind to other subtypes of 5-HT receptor. Digoxigenin-labeled oligonucleotides were found to detect mRNA encoding the 5 HT1A receptor in a subset of neurons in myenteric and submucosal ganglia. In contrast, 5-HT1A mRNA was not found in the pancreas. Radioautography revealed that the highest density of 3H-8-OH-DPAT binding sites was found in the stomach. These sites were especially numerous in the lamina propria adjacent to gastric glands, and in myenteric ganglia. Pancreatic 5-HT1A receptors were located on nerves, lymphoid tissue (especially the capsule of nodes), and on cells scattered in the pancreatic parenchyma. The concentration of 3H-8-OH-DPAT binding sites in the rat bowel and pancreas was less than that of 3H-5-HT binding sites; however, the distribution of 3H-8-OH-DPAT binding sites was similar to that of sites that bind 3H-5-HT. It is concluded that the rat gut and its extension in the pancreas contains 5-HT1A receptors. Many, if not all, of the nerve cells and processes that express 5-HT1A receptors express 5-HT1P receptors as well. The function of these receptors in the physiology of the entero-pancreatic innervation remains to be determined. PMID- 8425945 TI - Ultrastructure of the circuit providing input to the crayfish lateral giant neurons. AB - Labeled or otherwise identified neurons of the crayfish lateral giant escape reaction circuit were examined electron microscopically and the findings compared to expectations from physiology. Terminals of primary afferents contained clear, approximately 45 nm, irregularly round synaptic vesicles, while sensory interneuron terminals had slightly larger, 50 nm, more strictly round vesicles, permitting tentative classification based on anatomical criteria. Excitatory synapses on the lateral giants, believed from physiology to be electrical, generally had some gap junctions, but these were almost invariably paralleled by more prominent chemical junctional regions of unknown function. There may also be a class of interneurons making purely chemical synapses on the lateral giants. Synapses from primary afferents to sensory interneurons, believed from physiology to be cholinergic, had purely chemical morphology. Synapses with narrow elongated vesicles, similar to GABAergic vesicles seen in other neurons, frequently occurred on terminals of primary afferents. These synapses provide a basis for known presynaptic inhibition of afferent input. Consistent with physiology, such inhibitors sometimes also contacted the postsynaptic targets of the primary afferents and sometimes received input from other primary afferents. Afferent terminals also received some input from profiles rich in large dense cored vesicles. Presumptive inhibitory input found on proximal dendrites of lateral giants provides a basis for known recurrent inhibition. However, similar inhibitory synapses that sometimes received local input from excitors of the lateral giants were also found distally mixed with excitatory inputs. These provide a basis for recently discovered distal inhibitory input following excitation and for tonic inhibition. PMID- 8425946 TI - Electron microscopic immunocytochemical study of the distribution of parvalbumin containing neurons and axon terminals in the primate dentate gyrus and Ammon's horn. AB - Five green monkeys were examined with light and electron microscopic preparations to explore the regional differences in the distribution of parvalbumin (PV) positive neurons and axon terminals in the primate hippocampus. PV-positive neurons were mainly found in the hilus of the dentate gyrus and the strata oriens and pyramidale of Ammon's horn. In electron microscopic preparations, the PV positive cells displayed nuclear infoldings, intranuclear rods, a large rim of perikaryal cytoplasm with numerous organelles and both asymmetric and symmetric axosomatic synapses. One prominent PV-positive cell type in CA1 was a large multipolar neuron that resembled the large basket cells of the neocortex. Although most PV-positive dendrites were aspiny and postsynaptic to numerous axon terminals, some PV-positive dendrites in the molecular layer of the dentate gyrus displayed filipodia-like appendages with no synapses or spines that were postsynaptic to multiple axon terminals. The PV-positive dendrites in the hilus and stratum oriens were apposed at specialized junctions that resembled gap junctions. PV-positive axons were concentrated in the principal cell layers, and formed axosomatic, axodendritic, and axon initial segment synapses. In cases where these axons were observed to appose the surface of granule cells for a long length, only one axosomatic symmetric synapse per cell was found. In the hilus, PV-positive axon terminals formed synapses onto thorny excrescences of spiny cells. Both semithin sections and electron microscopic preparations indicated that more PV-positive axon terminals formed symmetric axosomatic synapses with pyramidal cells in CA2 than in CA1 and CA3. Also, CA2 displayed a unique plexus of PV-positive axon terminals in stratum lacunosum moleculare. These results indicate that the PV-positive hippocampal cells form a subset of GABAergic local circuit neurons, including the basket and chandelier cells. The ubiquitous finding of PV-positive dendrites linked by gap junctions throughout the dentate gyrus and Ammon's horn adds further data to indicate that this subset of GABAergic neurons is linked electrotonically. The synaptic organization of PV positive neurons in the hippocampus suggests their participation in both feedback and feedforward inhibition. The PV-positive neurons in the hippocampus are only a proportion of the basket and chandelier cells, whereas virtually all of these cells in neocortex are PV-positive. PMID- 8425947 TI - Cutaneous markers of internal malignancy. I. Malignant involvement of the skin and the genodermatoses. AB - Cutaneous findings often reflect the presence and course of an internal disease. Recognition of external clues is important to facilitate both early diagnosis and prompt treatment of the internal disorder. Early recognition is especially valuable in a patient with an internal malignant disease because intervention may significantly affect survival. In this two-part series, we review the spectrum of cutaneous markers of internal malignancy. Part I focuses on malignant involvement of the skin caused by either direct extension or metastases and the genodermatoses with malignant potential. Part II will be devoted to the paraneoplastic skin manifestations of internal malignancy as well as environmental carcinogens that produce cutaneous features. We also discuss some proposed but controversial associations between skin disease and internal malignancy. PMID- 8425948 TI - The association of potentially lethal neurologic syndromes with scleromyxedema (papular mucinosis). AB - Two patients with scleromyxedema who had progressive neurologic impairment are described. One patient died, and one required prolonged mechanical ventilation. A review of the literature has produced 24 other cases of scleromyxedema in which neurologic changes were prominent. PMID- 8425949 TI - Staphylococcal botryomycosis and hyperimmunoglobulin E (Job's) syndrome in an infant. PMID- 8425950 TI - Can chemicals precipitate dermatitis herpetiformis? PMID- 8425951 TI - False-positive Lyme antibody test in morphea. PMID- 8425952 TI - Granulomatous leg ulcers: an unusual presentation of Crohn's disease in a young man. PMID- 8425953 TI - Generalized pruritus: when to investigate further. PMID- 8425954 TI - No HLA antigen is significant in classic Kaposi's sarcoma. PMID- 8425955 TI - Boxing-glove hand: an unusual presentation of dermatitis artefacta. PMID- 8425956 TI - Unresponsive severe generalized pemphigus vulgaris successfully controlled by extracorporeal photopheresis. PMID- 8425957 TI - Erythrodermic bullous pemphigoid. PMID- 8425958 TI - Once-weekly treatment with oral ketoconazole for superficial fungal infections. PMID- 8425959 TI - Minocycline in Lyme disease. PMID- 8425960 TI - Subcutaneous scalp lesions in children. PMID- 8425961 TI - Erythema gyratum repens without underlying disease. PMID- 8425962 TI - Geriatric nail disorders. PMID- 8425963 TI - Topical cyclosporine for cicatricial pemphigoid. PMID- 8425964 TI - Calciphylaxis in patients with human immunodeficiency virus infection and renal disease. PMID- 8425965 TI - Erythrodermic dermatomyositis. PMID- 8425966 TI - Risk of another basal cell carcinoma developing after treatment of a basal cell carcinoma. AB - BACKGROUND: There is an increased risk of new basal cell carcinomas (BCCs) developing in a person who has had a BCC. OBJECTIVE: This study attempts to define the magnitude of this increased risk. METHODS: The charts of 260 white patients with a histologically proven BCC were reviewed for the occurrence of new BCCs. The cumulative 5-year incidence (modified life-table method) for new BCCs developing in these patients was compared with the 5-year incidence in the general white population of the United States. RESULTS: Of the 260 patients, new BCCs developed in 137 within an average of 38.3 months, a 5-year cumulative rate of one or more new BCCs of 45.2%. The yearly risk for new BCCs developing in the study population remained high during the 5-year interval. In the general white population of the United States, the maximal 5-year incidence was calculated to be 5% (p < 0.005, chi-square test). CONCLUSION: Patients with a history of BCC require life-long follow-up because of the high probability of new BCCs developing. PMID- 8425967 TI - Reflex sympathetic dystrophy: occurrence of chronic edema and nonimmune bullous skin lesions. AB - BACKGROUND: Reflex sympathetic dystrophy (RSD) is a poorly understood syndrome of post-traumatic pain, autonomic dysfunction, and progressive tissue atrophy. Classical descriptions of the cutaneous manifestations of RSD are usually limited to skin atrophy, vascular instability, and hyperhidrosis. OBJECTIVE: Our objective was to further delineate the cutaneous changes in RSD. METHODS: We have observed RSD-related inflammatory and bullous lesions in nine patients with active RSD. RESULTS: Eight patients had significant edema of involved skin, two patients had evidence of a pigmented purpura-like inflammatory dermatitis, and two other patients had bullae on involved skin. Ultrastructural studies on a biopsy specimen from one patient with recurrent bullae revealed a disrupted basement membrane and abnormal anchoring fibrils. CONCLUSION: Skin disease in RSD is more diverse than commonly appreciated and includes severe edema, inflammatory lesions, and a nonimmune bullous eruption. PMID- 8425968 TI - Labial melanotic macule: a clinical, histopathologic, and ultrastructural study. AB - BACKGROUND: The labial melanotic macule (LMM) is a recently described pigmentary anomaly that may simulate malignant melanoma. OBJECTIVE: Our purpose was to define the LMM clinically, histologically, and immunohistochemically in a large group of patients. METHODS: We describe the clinical features of 36 LMMs in 29 patients (aged from 4 to 79 years, 4 male, 25 female) seen during the past 4 years. Histopathologic findings in 21 of these patients are discussed. Seventeen lesions were immunostained with HMB-45 monoclonal antibody, and electron microscopy was performed on eight lesions. RESULTS: The majority of patients were women and had solitary lesions on the lower lip with the mean age of onset of 30 years. Histologically prominent basilar hyperpigmentation accentuated at the tips of the rete ridges was present without atypia or nevoid formation. Immunohistochemical studies showed that all intralesional melanocytes were HMB-45 negative, supporting their benign nature. Ultrastructurally, numerous stage III and IV melanosomes clustered within basal keratinocytes and papillary dermal melanophages were found. CONCLUSION: The LMM is a clinically and histologically distinctive benign pigmentary anomaly. PMID- 8425969 TI - Gamma delta T-cell receptor-positive cells in human skin. I. Incidence and V region gene expression in granulomatous skin lesions. AB - BACKGROUND: There have been many reports that gamma delta T-cell receptor (TCR)+ cells respond to mycobacterial antigens in vitro, but there is little available information on human gamma delta TCR+ cells in clinical conditions. OBJECTIVE: Our purpose was to investigate the distribution and involvement of human gamma delta TCR+ cells in granulomatous skin lesions. METHODS: The incidence and V region gene expression of human gamma delta TCR+ cells was examined in granulomatous skin diseases, including cutaneous tuberculosis and leprosy, by immunohistochemical procedures. RESULTS: gamma delta TCR+ cells in the dermis were increased in most patients with borderline lepromatous leprosy, and they were less frequently found in lepromatous leprosy and erythema nodosum leprosum. Other granulomatous skin lesions, including sarcoidosis, contained only a few gamma delta TCR+ cells. The gamma delta TCR+ cells that were found to be increased in this study were mostly delta TCS1-, BB3+, Ti gamma A+ (V delta 1-, V delta 2+, V gamma 9+). CONCLUSION: The gamma delta TCR+ cells in human granulomatous skin lesions may respond to some mycobacterial antigens, but they do not appear to be directly involved in granuloma formation. PMID- 8425970 TI - Excess melanocytic nevi in children with renal allografts. AB - BACKGROUND: Renal allograft transplantation is associated with an increased incidence of malignant melanoma. The development of excess melanocytic nevi may be an indicator of this risk. OBJECTIVE: This study determines the prevalence of melanocytic nevi in children who have received renal allografts. METHODS: Total and regional melanocytic nevi counts were made in 38 children (27 boys, 11 girls) with a renal allograft and in 38 individually age- and sex-matched healthy controls; counts were related to age, sex, skin type, and duration of immunosuppression. RESULTS: There was a significant increase in the total number of nevi in the renal transplant group compared with the control group (p < 0.05), with most marked increases occurring on the back and at acral sites. A strong positive correlation between nevi count and duration of immunosuppression independent of age was observed (p < 0.005). CONCLUSION: Excess numbers of melanocytic nevi occur in children with renal allografts. These patients constitute a risk group for malignant melanoma and require continued assessment. PMID- 8425971 TI - Autoeczematization is associated with abnormal immune recognition of autologous skin antigens. AB - BACKGROUND: The pathogenesis of autoeczematization (AE) is not well understood; however, previous studies suggest that AE is an autoimmune condition. OBJECTIVE: Our purpose was to assess whether AE is associated with an abnormal immune recognition of autologous skin antigens. METHODS: Eight patients with AE, six healthy control subjects, and three patients with localized contact dermatitis (LCD) were studied. Activation markers were detected on peripheral blood T lymphocytes. Autologous mixed epidermal cell-lymphocyte reaction (AMECLR) was performed for each subject and cell proliferation was assessed by tritiated thymidine incorporation. RESULTS: Many activated T cells were detected in patients with AE (5.2% +/- 4.5% vs 0.2% +/- 0.4% in control subjects, p < 0.05). AMECLR showed a significantly higher cell proliferation in AE compared with both healthy subjects and patients with LCD (6372 +/- 3217 cpm vs 2638 +/- 1788 cpm, and 2471 +/- 1389 cpm, respectively; p < 0.05). Peripheral blood mononuclear cells cultured in the presence of an autologous skin homogenate also showed a significantly increased cell proliferation in patients with AE than in control subjects. CONCLUSION: Our results suggest that an abnormal immune response against autologous skin antigens occurs in AE that could be related to the pathogenesis of this disease. PMID- 8425972 TI - Cutaneous side effects associated with interleukin 2 administration for metastatic melanoma. AB - BACKGROUND: Cutaneous changes associated with recombinant interleukin 2 (r IL-2) administration are frequent but have been rarely studied in a large series. OBJECTIVE: We analyzed these clinical, microscopic, and immunologic changes. METHODS: Patients with metastatic melanoma treated with r IL-2 were studied. The eruption was scored as mild or severe. Biopsy specimens were obtained for histopathology, ultrastructural analysis, and immunophenotyping. Chi-square and t tests were used for statistics. RESULTS: Twenty-five patients were included. Eruptions were observed in 56 of 78 cycles (72%); 53 were mild with a burning pruriginous erythema, and 3 were severe with urticaria, necrotic lesions, and blisters. Regression was constant without sequelae. Pathologic changes were mild with a mononuclear cell infiltrate of activated helper T phenotype, expressing LFA-1. Keratinocytes and endothelial cells displayed intercellular adhesion molecule-1 and HLA-DR. Cells rarely expressed CD25. CONCLUSION: Administration of r IL-2 triggers non-treatment-limiting cutaneous inflammation. PMID- 8425973 TI - Stanozolol causes rapid pain relief and healing of cutaneous ulcers caused by cryofibrinogenemia. AB - BACKGROUND: Cutaneous manifestations of cryofibrinogenemia include purpura, ecchymosis, and ulcerations. The histology of these lesions is characterized by intravascular thrombi. OBJECTIVE: Our purpose was to test the efficacy of stanozolol, a drug capable of fibrinolytic enhancement, in treating cutaneous ulcers caused by cryofibrinogenemia. METHODS: Eight patients with cutaneous ulcerations from cryofibrinogenemia were treated with stanozolol. Plasma cryofibrinogen was measured before and during treatment with stanozolol. Histologic evaluation was also performed before treatment and in selected patients during treatment. RESULTS: After treatment, seven of the eight patients had healing of their ulcers, prompt reduction in their pain, and improvement in livedo reticularis and purpura. Four of the eight patients had no detectable plasma cryofibrinogen after treatment. In addition, dermal intravascular thrombi resolved. Stanozolol was well tolerated and had minimal side effects. CONCLUSION: We conclude that stanozolol is a safe and effective treatment of the cutaneous manifestations of cryofibrinogenemia. PMID- 8425974 TI - Treatment of generalized bullous pemphigoid with oral tetracycline. AB - BACKGROUND: Although bullous pemphigoid (BP) is a benign self-limited disease, the mainstay of treatment remains systemic steroids, often in combination with immunosuppressive agents. This therapy has considerable potential toxicity, particularly in elderly patients with preexisting problems. OBJECTIVE: The purpose of this study was to evaluate the efficacy of oral tetracycline as first choice therapy in patients with BP. METHODS: Every patient newly diagnosed with generalized BP was treated with oral tetracycline and a midpotency topical steroid. RESULTS: In all five patients, blister formation was stopped and reepithelialization completed within 1 to 3 weeks. There was no relapse or toxicity noted; follow-up ranged from 16 to 24 months. CONCLUSION: Oral tetracycline was found to be rapidly efficacious in all patients and devoid of toxicity. PMID- 8425975 TI - A radical proposal for the pathogenesis of scleroderma. AB - Because the etiology and pathogenesis of scleroderma are largely speculative, effective therapy is not available for this debilitating condition and its variants. This article reviews current concepts in the etiology and pathogenesis of scleroderma and suggests a unifying hypothesis involving the generation of free radicals. The role of specific free radical scavengers should be tested both in vitro and in clinical trials of patients with scleroderma. PMID- 8425976 TI - Focal dermal hypoplasia (Goltz syndrome): an adult case with multisystemic involvement. AB - Focal dermal hypoplasia (Goltz syndrome) is a rare congenital syndrome with suspected X-linked transmission that is characterized by a wide range of mesoectodermal defects. We describe a 39-year-old woman who had a peculiar phenotype and asymmetry of the body. Examination revealed atrophic erythematous and hyperpigmented linear streaks following Blaschko's lines, some of which were in a reticular or cribriform arrangement. Yellow, soft nodules caused by fat herniation were visible mainly in skin folds. The patient had multiple bone anomalies, including longitudinal striation of long bones (osteopathia striata). She had several ocular, dental, and kidney defects. Histopathologic examination showed a markedly thinned dermis that was replaced by adipose tissue. PMID- 8425977 TI - Brunsting-Perry cicatricial bullous pemphigoid: a clinical variant of localized acquired epidermolysis bullosa? AB - An 84-year-old man who had the typical clinical features of Brunsting-Perry cicatricial pemphigoid is described. Direct immunofluorescence microscopic examination of salt-split skin revealed linear deposits of IgG and C3 on the floor of the artificial bullae. Direct immunoelectron microscopic examination of peribullous skin showed dermal cleavage level below the lamina densa and granular deposits of IgG and C3 attached to and below the lamina densa in a pattern identical to epidermolysis bullosa acquisita. These findings suggest that Brunsting-Perry cicatricial pemphigoid may represent a clinical variant of epidermolysis bullosa acquisita. PMID- 8425978 TI - Sezary syndrome in an 11-year-old girl. AB - All forms of cutaneous T-cell lymphoma are rare in children. We describe an 11 year-old girl who had generalized exfoliative erythroderma, intense pruritus, peripheral lymphadenopathy, mycosis cells in the skin and lymph nodes, and Sezary cells in the peripheral blood. Results of a biopsy specimen of involved skin showed changes consistent with mycosis fungoides. A classic case of Sezary syndrome has not previously been reported in childhood or preadolescence. PMID- 8425979 TI - Erosive pustular dermatosis of the scalp. AB - Three cases of erosive pustular dermatosis of the scalp are reported. In all patients the dermatosis was characterized by pustular, erosive, and crusted lesions; in addition, two patients had areas of scarring alopecia. The results of laboratory tests, bacteriologic and mycologic investigations, and histopathologic examination were nondiagnostic. Although erosive pustular dermatosis of the scalp is characterized by a nonpathognomonic clinical and histopathologic picture, it probably represents a disease entity. PMID- 8425980 TI - Hepatonecrosis resulting from parenteral gold therapy in pemphigus vulgaris. AB - A patient who had pemphigus vulgaris received therapy with gold thiomalate in addition to dexamethasone. Three weeks after the initiation of parenteral gold the patient developed acute hepatitis. Causes other than gold toxicity were ruled out, and the patient recovered completely after discontinuation of gold therapy. This is the first case of acute hepatonecrosis resulting from gold therapy of pemphigus to be reported in the dermatologic literature. PMID- 8425981 TI - Composite resin attached to glass polyalkenoate (ionomer) cement--the laminate technique. AB - Since the introduction of the laminate technique where composite resin is placed over glass polyalkenoate cement in an endeavour to obtain a marginal seal to tooth substance, it has been advocated and used by many clinicians. This review examines the available literature on this apparently commonly used technique. A wide-ranging evaluation of the technique is allowed by the extensive nature of the literature. The conclusion of the review is that more work is required to evaluate the procedure, but the results of in vitro and, in particular, in vivo studies suggest that the technique may not be as successful clinically as the theory may support. PMID- 8425982 TI - Marginal adaptation and fit of adhesive ceramic inlays. AB - This in vitro study compared the marginal adaptation of CAD/CAM and laboratory made ceramic inlays before, during and after loading. Six MOD inlay preparations of standardized design with one cervical margin in dentine and the other in enamel were prepared for each inlay type: CAD/CAM fabricated MGC-glass ceramic inlays, CAD/CAM fabricated feldspathic porcelain inlays, laboratory-made glass ceramic inlays and laboratory-made feldspathic porcelain inlays. Appropriate luting composite materials were used. The restored teeth were subjected to occlusal loading, thermal cycling, toothbrush-toothpaste abrasion and chemical degradation in vitro. Marginal adaptation was quantitated along the entire length of the cavosurface margin and along selected sections of the margin using SEM, following in vitro testing corresponding to 0, 0.5, 1.0, 2.7 and 5.0 years of clinical service. In addition, marginal fit of the cemented inlays was evaluated in the SEM. The initial marginal adaptation in enamel was excellent in all groups. After in vitro testing, significant marginal discrepancies were found in all groups. A high percentage of marginal openings was recorded, notably in the cervical portions of the margins in both enamel and dentine. PMID- 8425983 TI - In vitro comparison of conventional crowns and a new all-ceramic system. AB - Thirty-three crowns of similar size and shape were constructed to fit a standard brass die. Eleven crowns were manufactured from each of the following groups of materials: metal-ceramic, aluminous porcelain and a new all-ceramic system based on a glass infiltrated alumina coping. The overall fitting accuracy of each crown was determined using a low viscosity silicone paste, prior to cementation onto brass dies with zinc phosphate cement. The average film thickness was 95, 154 and 123 microns respectively. Each cemented crown was loaded to fracture. The mean load to failure of the glass infiltrated alumina crowns was significantly greater than that of the aluminous porcelain jacket crowns (P < 0.0001), but not significantly different from the metal-ceramic crowns. PMID- 8425984 TI - Dental materials: 1991 literature review. PMID- 8425985 TI - Effect of a resin lining and rebonding on the marginal leakage of amalgam restorations. AB - This in vitro study compared microleakage in Class V amalgam restorations with three different lining agents and with no lining agent, with and without resin rebonding of the margins. Eight groups of ten molars were divided into four pairs of groups. Class V preparations were cut in the facial surfaces of each molar so that the occlusal margin was in enamel and the gingival margin in dentine. The preparations in one pair of groups received no lining agent; the preparations in the remaining three pairs of groups were lined with one of the following: Copalite, Universal Bond 3 (UB3) Primer and Adhesive, or UB3 Primer only. The enamel walls of the preparations receiving the UB3 Primer and Adhesive were etched prior to applying the lining material. Amalgam was hand condensed into each preparation. In one of each pair of groups, the margins of restorations were treated with 37% phosphoric acid gel, rinsed and dried, and UB3 Adhesive was applied over amalgam and tooth margins and polymerized (rebonded). Specimens were thermocycled, stained and sectioned. Microleakage was graded using a stereomicroscope. Mean microleakage scores for occlusal and gingival margins were calculated and analysed. At enamel margins, non-rebonded Copalite and all rebonded specimens showed less microleakage than the other non-rebonded groups. The group lined with UB3 Primer only and rebonded showed significantly less (P < 0.01) microleakage at enamel margins than all the other groups except the group lined with Copalite and rebonded. At cementum/dentine margins, restorations lined with UB3 Primer and rebonded showed significantly less microleakage than the other groups. PMID- 8425986 TI - Finite element analysis of quasistatic and fatigue failure of post and cores. AB - Finite element (FE) analysis of the mechanical behaviour of materials and structures facilitates the investigation of their internal stress distributions. However, the validity of the model is not always ascertained. In this study a three-dimensional (3D) FE model was developed, representing a laboratory set-up of direct post and core restored upper premolars. These restorations, using either composite or amalgam for core material, have been the subject of study in previous quasistatic and fatigue strength tests. The aim of this study was to validate the FE model for prefailure and failure modelling, by comparing the computational results with the laboratory observations and failure results. Two failure criteria were selected for investigation: Modified Von Mises and Drucker Prager equivalent stress. Four model variations were carried out, representing different conditions at the core-tooth interface. Prefailure modelling was found to be adequate. The calculated failure results could only partly be fitted to the quasistatic tests. The best fit was effected with a model using partial bonding of the core, for the composite core. Fatigue failure was reproduced somewhat better by a model using no bonding at all, again to a higher degree for the composite core. Calculations of post stress using a model simulating increased core mobility supported an observation made previously (M. C. D. N. J. M. Huysmans et al., in press; Int. Endodont. J. XX, XXX-XXX), implying that a composite core raises the demands made on the post. The conclusion is made that validation of FE calculations is essential. A 3D model as presented here shows a satisfactory fit to fatigue data but not to quasistatic results.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8425987 TI - Comparative case fatality analysis of the International Tissue Plasminogen Activator/Streptokinase Mortality Trial: variation by country beyond predictive profile. The Investigators of the International Tissue Plasminogen Activator/Streptokinase Mortality Trial. AB - OBJECTIVES: This study was designed to examine the variation in mortality rates among countries participating in the International Tissue Plasminogen Activator/Streptokinase Mortality Trial. BACKGROUND: Despite uniform inclusion and exclusion criteria and protocol in this trial, 30-day mortality rates (irrespective of treatment allocation) ranged from 4.2% to 14.8% among the participating countries. METHODS: With use of the risk factors identified by a multi-variate logistic model, the total study group was classified into deciles on the basis of each patient's risk profile and individual probability of dying within 30 days. Expected mortality rates were then calculated and compared with actual mortality for each decile of the total study group, as well as for patients from each country. RESULTS: Independent risk factors for mortality were older age (odds ratio 1.97 for each 10-year increment), systolic hypotension (blood pressure < 95 mm Hg) at entry (odds ratio 3.7), Killip class > 1 at entry (odds ratio 3.5), history of antecedent angina (odds ratio 1.23 to 1.49), history of diabetes mellitus (odds ratio 1.64), previous infarction (odds ratio 1.23) and history of never smoking (odds ratio 1.37). The overall mortality rate among the 1,612 patients in risk deciles 9 and 10 was 26%; for the 1,606 patients in deciles 1 and 2 it was 1.2%, with a sensitivity of 58.6% and a specificity of 83.7%. The logistic model closely predicted and explained the different mortality rates for most countries (the differences between expected and actual mortality were nonsignificant). However, in the total study group, the difference between the expected and actual mortality was significant (p < 0.001). This difference was mainly ascribed to the two countries with the highest and lowest mortality rates. When the patients from these two countries were excluded from the analysis, the overall difference became nonsignificant. CONCLUSIONS: These findings suggest that the recognized risk factors associated with increased case fatality in acute myocardial infarction account only in part for mortality differences across or within populations. PMID- 8425988 TI - Coronary rotational ablation: initial experience in 302 procedures. AB - OBJECTIVES: The aim of this study was to assess the utility of percutaneous transluminal coronary rotational ablation in the treatment of coronary artery disease. BACKGROUND: Although numerous advances have been made in the treatment of coronary artery disease, there are lesions with complex morphology that are not amenable to current intravascular therapy. METHODS: A consecutive series of 242 patients having 302 coronary rotational ablation procedures was analyzed. One hundred nineteen (49%) of the patients had previously undergone attempted coronary angioplasty, which was unsuccessful in 31 patients (13%). The left ventricular ejection fraction was normal in 196 patients (81%). The ablation procedure was attempted in 308 vessels and 346 lesions. Of the 346 lesions treated, 26 (7.5%) were classified as American College of Cardiology/American Heart Association type A, and 320 (92.5%) as either type B or type C. RESULTS: Procedural success was achieved in 284 (94%) of the 302 procedures and 330 (95.4%) of the 346 lesions in which ablation was attempted. Five procedures (1.7%) were unsuccessful, but no cardiac event occurred during the hospital stay. A major cardiac event occurred in 13 cases (4.3%); 9 (3%) of these complications were due to the ablation procedure. Six patients sustained a Q wave myocardial infarction alone, two had a Q wave infarction and required emergency surgery and one needed emergency surgery but did not have a Q wave infarction. No procedural deaths were attributed to the ablation procedure. Follow-up has been obtained in 182 of the 242 patients at a mean interval of 9 +/- 5 months. Of the 182 patients, 174 (95.6%) were alive and free of myocardial infarction. Angiographic follow-up is available thus far in 87 patients. By combining angiographic and clinical outcome, an overall estimated restenosis rate of 37.4% (68 of 182) was calculated. CONCLUSIONS: These data suggest that coronary rotational ablation can be performed on lesions with a variety of morphologic features with high initial success rates. The overall rate of restenosis is similar to that of balloon angioplasty. PMID- 8425989 TI - Rotational coronary ablation: more grist for the interventional mill? PMID- 8425990 TI - Intracoronary ultrasound assessment of directional coronary atherectomy: immediate and follow-up findings. AB - OBJECTIVES: This study was conducted to assess the relations among intracoronary ultrasound, angiographic and histologic data obtained from patients with coronary artery disease successfully treated by directional coronary atherectomy. In addition, it was designed to elucidate whether some aspects of intravascular ultrasound or pathologic findings could predict a propensity to restenosis. BACKGROUND: Intracoronary ultrasound is a useful technique in guiding and assessing atherectomy. However, there is little information about the characterization of the different types of coronary plaques and the changes observed in them after resection. Furthermore, the follow-up ultrasound appearance of previously treated lesions remains undepicted. METHODS: Fifty-two patients (54 +/- 10 years old) were studied. All were successfully treated by atherectomy with the aid of intracoronary ultrasound guidance. Qualitative and quantitative ultrasound and angiographic variables were derived before and after resection. Quantitative histologic morphometric information was also obtained from the specimens. In 22 patients, a follow-up echoangiographic reevaluation was performed 6 +/- 4 months later. RESULTS: Echogenic plaques had a higher collagen and calcium content, whereas echolucent plaques had an increased level of fibrin, nuclei and lipids. Ultrasound plaque reduction after atherectomy was greater in echolucent (76 +/- 21%) than in echogenic plaques (60 +/- 18%; p < 0.05). That reduction correlated with the weight of the resected material (r = 0.62; p < 0.01). At follow-up study, 13 of 22 patients had angiographic and ultrasound evidence of restenosis. Most recurrent lesions had a stenotic three-layer appearance. The incidence of restenosis of primary lesions treated with atherectomy was higher in echolucent (100%) than in echogenic (33%) plaques. Similarly, a higher proportion of nuclear content in the resected material was observed in patients who developed restenosis (2.1 +/- 0.7%) than in patients who had late success after atherectomy (1.2 +/- 0.6%). CONCLUSIONS: Our findings suggest that echolucent plaques are easier to resect than are echogenic plaques but frequently develop restenosis. In contrast, the resection of echogenic plaques, although often incomplete, is associated with better long-term results. PMID- 8425991 TI - Analysis of coronary blood flow velocity dynamics in angiographically normal and stenosed arteries before and after endolumen enlargement by angioplasty. AB - OBJECTIVES: This study was designed to assess whether the spectral waveform of coronary velocity on Doppler study is characteristically altered in the presence of significant stenosis with normalization of the spectral waveform after relief of endolumen obstruction. BACKGROUND: Although coronary flow reserve determinations have provided physiologic information complementary to the angiographic percent diameter narrowing, flow velocity measurements have been limited to proximal arteries with inconsistent results after angioplasty. A 12 MHz Doppler guide wire permits flow velocity determination in the proximal and distal coronary artery with fast Fourier spectral analysis. METHODS: With the Doppler guide wire, proximal arterial flow velocity and flow reserve measurements in 17 angiographically normal arteries were compared with measurements in 29 significantly stenosed arteries. Proximal and distal flow velocity measurements were also obtained before and after angioplasty of the 29 abnormal arteries. Velocity spectrum was digitized to compute peak diastolic velocity, peak systolic velocity, mean velocity, diastolic/systolic velocity ratio and first third and first half flow fraction. RESULTS: Compared with proximal stenosed arteries, proximal normal arteries had significantly higher peak diastolic velocity (64 +/- 26 cm/s vs. 41 +/- 26 cm/s) and higher coronary vasodilator reserve (2.3 +/- 0.8 vs. 1.6 +/- 0.7). Normal arteries had higher flows in the first third and first half of the coronary cycle (46 +/- 3% vs. 39 +/- 7% and 65 +/- 2% vs. 56 +/- 10%, respectively). Before angioplasty, coronary velocity variables were significantly lower distal than proximal to the stenosis. After angioplasty, there was a greater mean increase in distal velocities (200% vs. 90% for the proximal arteries) that resulted in near equalization of proximal and distal mean velocity and a significant reduction in proximal/distal mean velocity ratio (2.4 +/- 1.7 vs. 1.2 +/- 0.4). CONCLUSIONS: Before angioplasty, abnormal coronary flow velocity dynamics are more marked distal than proximal to the stenosis. Greater increase in coronary flow velocities in the distal circulation after relief of endolumen obstruction results in a significant reduction in the proximal/distal flow velocity ratio. Thus, normalization of Doppler-derived flow velocity variables with marked reduction of the proximal/distal flow velocity ratios parallels angiographic success and may prove useful as an additional end point measurement in interventional cases with questionable angiographic findings. PMID- 8425992 TI - Which angiographic variable best describes functional status 6 months after successful single-vessel coronary balloon angioplasty? AB - OBJECTIVES: The aim of this study was to determine which quantitative angiographic variable best describes functional status 6 months after coronary balloon angioplasty. BACKGROUND: Several angiographic restenosis criteria have been developed. These can be divided into those that describe the change in lesion severity and those that merely describe lesion severity at follow-up angiography. The functional significance of these criteria is unknown. METHODS: We studied 350 patients with single-vessel coronary artery disease who underwent a single-site balloon dilation. Sensitivity and specificity curves were constructed for the prediction of anginal status and exercise electrocardiography of four quantitative angiographic variables that describe restenosis. The point of highest diagnostic accuracy for the variables was determined at the intersection of the sensitivity and specificity curves. Results of exercise electrocardiography were considered indicative for ischemia 6 months after angioplasty if horizontal or downsloping ST segment depression > or = 1 mm occurred. RESULTS: The points of highest diagnostic accuracy of the angiographic variables were similar for both anginal status and exercise electrocardiography: 1.45 and 1.46 mm for the minimal lumen diameter measurements, 45.5% and 46.5% for the percent diameter stenosis measurements at follow-up, -0.30 and -0.32 mm for change in minimal lumen diameter and -10% and -10% for the change in percent diameter stenosis at follow-up. CONCLUSIONS: Angiographic variables reflecting a change in lesion severity at follow-up angiography were only slightly less accurate than variables that describe lesion severity at follow-up. The large study group and the fact that the same optimal values for diagnostic accuracy of the various quantitative angiographic variables were obtained for the prediction of two different markers of ischemia suggests that these values reflect the lesion severity or increase in lesion severity in major epicardial vessels at which coronary flow reserve is unable to meet myocardial demands. PMID- 8425993 TI - Extent of jeopardized viable myocardium determined by myocardial perfusion imaging best predicts perioperative cardiac events in patients undergoing noncardiac surgery. AB - OBJECTIVES: This study was undertaken to test the hypothesis that the risk of perioperative cardiac events is not simply determined by the presence of myocardium at risk, but is directly related to the extent of myocardium at risk as reflected in thallium-201 myocardial imaging. BACKGROUND: The risk of perioperative cardiac events in patients undergoing noncardiac surgery has been related to the presence of transient defects on dipyridamole thallium-201 myocardial imaging, reflecting jeopardized viable myocardium. METHODS: The study cohort consisted of 231 consecutive patients who underwent noncardiac surgery and had a preoperative dipyridamole thallium-201 imaging study. Patients with vascular reconstruction or bypass constituted the largest surgical subgroup (n = 140). For thallium-201 imaging data, each of three planar projections was divided into three segments (total nine segments) and each segment was interpreted as normal or showing a transient or fixed defect. The ability of clinical and thallium-201 imaging data to predict perioperative cardiac events was compared with stepwise multivariate logistic regression analysis. RESULTS: Perioperative cardiac events occurred in 19 patients, including 5 with cardiac death, 7 with nonfatal myocardial infarction and 7 with unstable angina. For cardiac death or nonfatal myocardial infarction, the only significant multivariate predictors were the number of myocardial segments with transient thallium-201 defects (p < 0.0005) and a history of diabetes mellitus (p < 0.05). For all cardiac events, the only significant multivariate predictors were the number of myocardial segments with transient defects (p < 0.0001), diabetes mellitus (p < 0.05) and calcium channel blocker use (p < 0.05). CONCLUSIONS: The probability of important cardiac events in patients undergoing noncardiac surgery is best predicted by the extent of myocardium at risk as reflected on thallium-201 myocardial perfusion imaging. A history of diabetes mellitus also has a significant influence on perioperative risk. PMID- 8425994 TI - Relative importance of psychologic traits and severity of ischemia in causing angina during treadmill exercise. Canadian Amlodipine/Atenolol in Silent Ischemia Study (CASIS) Investigators. AB - OBJECTIVES: This study was conducted to compare the influence of psychologic traits versus ischemia severity on the occurrence of angina during treadmill exercise. BACKGROUND: Some studies suggest that angina is associated with certain psychologic traits, whereas others show an association with more severe ischemia. The relative influence of these two factors and the extent to which they interact are not known. METHODS: Off-drug treadmill exercise testing and a battery of psychologic tests were performed on 122 patients with known coronary artery disease. Psychologic tests measured sensitivity to physical symptoms, denial and deception, type A behavior, anger, hostility, depression, marital adjustment and amount of external stress. Stepwise logistic regression was used to determine the independent association of psychologic traits, ischemic threshold and exercise tolerance with the occurrence of angina. RESULTS: Angina during treadmill exercise was reported by 66 of 122 patients. On univariate testing, angina was positively associated with sensitivity to physical symptoms (p < 0.001), type A behavior (p = 0.021) and depression (p = 0.032) and was negatively associated with exercise tolerance (p < 0.001) and work load threshold for ischemia (p < 0.01). Multivariate analysis revealed independent and additive associations of angina with sensitivity to physical symptoms (p = 0.003), exercise capacity (p = 0.003) and work load threshold for ischemia (p = 0.018). Once these were included in a logistic model, depression and type A behavior were no longer significant. Other psychologic traits showed no association with angina. CONCLUSIONS: Sensitivity to physical symptoms, ischemic threshold and exercise tolerance are independently associated with angina, with sensitivity to physical symptoms having the stronger influence. The physiologic and psychologic mechanisms underlying symptom perception have an influence on angina that is independent of and additive to the severity of underlying ischemia. PMID- 8425995 TI - Transdermal nitroglycerin reduces the frequency of anginal attacks but fails to prevent silent ischemia. AB - OBJECTIVES: The aim of this study was to objectively evaluate the effects of intermittent administration of transdermal nitroglycerin on effort tolerance, frequency of anginal attacks and presence of silent ischemic events that occur during normal daily activities. BACKGROUND: Previous studies have shown that transdermal nitroglycerin patches reduce the incidence of anginal attacks and improve exercise capacity when given intermittently. However, no carefully controlled studies are available on the effects of these preparations (and their dosing schedule) on the occurrence of "silent" ischemic events during unrestricted daily activities. METHODS: Twelve men with chronic stable angina, a positive exercise test result and significant coronary artery disease completed a randomized, double-blind, placebo-controlled trial in which patches were worn either continuously or with overnight (8 h) removal. The effects of treatment were objectively assessed by both treadmill exercise testing and 24-h ambulatory electrocardiographic monitoring. RESULTS: Only the intermittent dosing schedule afforded a small but significant improvement in exercise tolerance and prolonged exercise duration and time to ST segment depression. The frequency of anginal attacks was also reduced by both the continuous and intermittent treatment, but the effects on symptoms were not paralleled by a concomitant reduction in ischemic episodes recorded during ambulatory monitoring. CONCLUSIONS: The results indicate that when used as monotherapy, intermittent transdermal nitroglycerin preparations lessen symptoms but are ineffective for the long-term prophylaxis of silent myocardial ischemia. PMID- 8425996 TI - Effect of increases in heart rate and arterial pressure on coronary flow reserve in humans. AB - OBJECTIVES: The objective of this study was to determine the effect of increases in heart rate and arterial pressure on maximal pharmacologic coronary blood flow reserve. BACKGROUND: Coronary flow reserve measurements are useful in assessment of the physiologic significance of coronary lesions. However, animal studies suggest that alterations in hemodynamic status may influence coronary flow reserve independent of coronary stenosis. METHODS: Coronary flow reserve was measured during cardiac catheterization with the use of a 3F coronary Doppler catheter and intracoronary papaverine. Flow reserve was measured under control conditions and during increases in heart rate produced by atrial pacing (18 patients) or during elevation of arterial pressure by intravenous phenylephrine infusion (9 patients) with intracoronary alpha-adrenergic blockade by phentolamine. RESULTS: Coronary flow reserve progressively decreased from 3.7 +/- 0.9 (mean +/- SD) at the rate of 71 +/- 8 beats/min at rest to 3.0 +/- 0.6 during pacing at 100 beats/min and to 2.6 +/- 0.5 during pacing at 120 beats/min. Flow reserve decreased because of a progressive increase in rest coronary flow velocity during pacing (122 +/- 16% of control value at 100 beats/min, 139 +/- 16% of control value at 120 beats/min), whereas papaverine hyperemia peak velocity remained unchanged. Flow reserve decreased with pacing tachycardia whether the initial flow reserve was normal or depressed. Mean arterial pressure increased from 95 +/- 12 mm Hg to 130 +/- 8 mm Hg during intravenous phenylephrine infusion and to 123 +/- 10 mm Hg during combined intravenous phenylephrine and intracoronary phentolamine infusions. Coronary flow reserve was not affected by the blood pressure increases (control value 4.3 +/- 1.0, phenylephrine 4.4 +/- 1.5, phenylephrine and phentolamine 4.4 +/- 2.0). CONCLUSIONS: Sudden increases in heart rate but not mean arterial pressure lead to a substantial reduction in maximal coronary blood flow reserve. These data suggest that the diagnostic utility of all flow reserve measurement techniques might be improved by standardization of heart rate during measurement or extrapolation of the measured flow reserve to that expected at a reference heart rate. PMID- 8425997 TI - Myocardial contrast echocardiography for the assessment of coronary blood flow reserve: validation in humans. AB - OBJECTIVES: The aim of this study was to validate the use of myocardial contrast echocardiography to determine coronary blood flow reserve in humans. BACKGROUND: Although myocardial contrast echocardiography has been used to accurately quantify coronary flow reserve in animals, validation for its use in humans to measure flow reserve is lacking. METHODS: We analyzed the time-intensity curve from the anteroseptal region of the left ventricular short axis produced after a left main coronary artery injection of sonicated albumin before and after intracoronary administration of papaverine in 16 patients without angiographically significant coronary artery disease. The ratio of half-time of video intensity disappearance from peak intensity, variable of curve width, area under the time-intensity curve and corrected peak contrast intensity after papaverine compared with baseline were correlated with coronary flow reserve measured simultaneously with an intracoronary Doppler probe in the left anterior descending coronary artery. RESULTS: There was a strong inverse correlation with half-time of contrast washout and coronary flow reserve (r = -0.76, p = 0.0007) and a strong positive correlation between the variable of curve width (which is inversely proportional to curve width) and coronary flow reserve (r = 0.71, p = 0.002). There was a weak but significant inverse correlation between area under the time-intensity curve and coronary flow reserve (r = -0.54, p = 0.03) but no correlation between corrected peak contrast intensity and coronary flow reserve (r = -0.36, p = NS). Despite the strong correlation for the ratios for half-time of contrast washout and variable of curve width and actual coronary flow reserve measured with intracoronary Doppler probe, the transit time ratios consistently underestimated coronary flow reserve. CONCLUSIONS: Myocardial contrast echocardiography performed with left main coronary artery injections of sonicated albumin can be utilized to measure coronary flow reserve in humans. Transit time variable ratios (half-time of contrast washout and variable of curve width) derived from the time-intensity curve correlate most strongly with coronary flow reserve. PMID- 8425998 TI - Myocardial contrast echocardiography has the potential for the assessment of coronary microvascular reserve. AB - Coronary vasodilators increase coronary flow by increasing myocardial blood volume. Diseases affecting the coronary microvasculature will affect vasodilator induced changes in coronary flow by inhibiting changes in myocardial blood volume. In such cases, when myocardial time-intensity curves after administration of a vasodilator are compared with those at baseline, a less than anticipated increase in microbubble transit rates will be noted. As long as we understand what we are measuring in the context of where and how we inject the bubbles, we can begin to define the role of myocardial contrast echocardiography in assessing changes in coronary microvascular reserve. It is also conceivable that because myocardial contrast echocardiography can assess changes in myocardial flow/volume relations rather than just changes in flow, this technique could be used to provide additional insights into the mechanisms of action of different coronary vasodilators and into the pathophysiology of various diseases affecting the coronary microvasculature. Finally, with the advent of commercially available microbubbles, robust on- and off-line analysis algorithms and intracardiac imaging, myocardial contrast echocardiography may become an invaluable adjunct to coronary angiography for determining the pathophysiologic significance of coronary disease in individual patients. PMID- 8425999 TI - Enhanced coronary blood flow velocity during intraaortic balloon counterpulsation in critically ill patients. AB - OBJECTIVES: The aim of this study was to assess coronary blood flow during intraaortic balloon counterpulsation by direct measurement. BACKGROUND: In a majority of human studies, increased coronary blood flow during intraaortic balloon counterpulsation measured by indirect techniques has not been consistently demonstrated. METHODS: Hemodynamic variables and coronary blood flow velocity (20-MHz Doppler-tipped catheter) data were measured in 19 patients requiring intraaortic balloon pumping for clinical indications (11 patients had acute myocardial infarction [9 with shock], 6 had unstable angina, 1 had acute mitral regurgitation and 1 was at high risk undergoing angioplasty). Hemodynamic data, mean and phasic diastolic flow velocity and velocity-time integrals (computed from digitized waveforms) were analyzed during periods of 1:1 balloon counterpulsation. RESULTS: Intraaortic balloon pumping decreased systolic pressure (6 +/- 10%, p < 0.001) and increased diastolic pressure (80 +/- 30% from baseline, p < 0.001) without changing RR interval. Peak phasic, mean coronary flow velocity and diastolic flow velocity integral were significantly increased (115 +/- 115%, 67 +/- 61%, 103 +/- 81%, respectively, all p < 0.001) during intraaortic balloon pumping. In addition, although a wide splay of data was evident due to operator set variations in balloon inflation and deflation timing, the greater increases in diastolic flow velocity integral (DFVi) occurred in patients with basal systolic pressure < or = 90 mm Hg (% delta DFVi = 102 - 0.1.[unaugmented systolic pressure], SEE = 21.7 mm Hg, r = 0.30, p < 0.001). CONCLUSIONS: Intraaortic balloon pumping unequivocally and significantly augments proximal coronary blood flow velocity, nearly doubling the coronary flow velocity integral in most patients. This mechanism may be a significant means of ischemia relief in hypotensive patients. PMID- 8426000 TI - Isolated left main coronary ostial stenosis in Oriental people: operative, histopathologic and clinical findings in six patients. AB - OBJECTIVES: This study was performed to determine whether there are differences in the operative, histopathologic, angiographic and clinical findings of isolated ostial stenosis between Oriental and western patients. BACKGROUND: Angiographic, clinical and histologic findings in isolated ostial stenosis have been reported in western but not in Oriental patients. METHODS: Six patients, all women (0.88% of a total of 684 patients who underwent coronary angiography between March 1989 and July 1991), were found to have isolated left main coronary ostial stenosis. We performed surgical ostial angioplasty with the autologous pericardial or saphenous venous patch and biopsy at the aortic arteriotomy site in four of the six patients. RESULTS: All six patients presented with severe angina (angina class III or IV) of short duration (mean +/- SD 6.2 +/- 6.2 months) and had a very low incidence of risk factors, although histopathologic examination showed typical atherosclerosis in four of the six patients. They were young to middle aged women (mean 45 +/- 3 years) except for Patient 6 (62 years). Exercise duration was short and ST segment depression, accompanied by typical angina, was observed in many leads in the warm-up period or stage I. Despite the crucial location of the lesion, most patients had well preserved left ventricular function and normal wall motion. There was no angiographically definable collateral circulation from either ipsilateral or contralateral vessels except for grade I collateral circulation in Patient 5. Operative findings demonstrated mostly yellow atheroma in the aortic wall and left coronary ostium. Coronary angiography showed only ostial stenosis of the left coronary artery in all six patients, but operative findings documented atheromatous change in the left main coronary artery in two of the six. CONCLUSIONS: The clinical, angiographic, histopathologic and operative findings of Oriental patients were similar to those reported in western patients, but the incidence of isolated left main coronary ostial stenosis was higher in the Oriental group. Angiographically definable isolated coronary ostial stenosis may often not be true isolated ostial stenosis. PMID- 8426001 TI - Noninvasive estimation of regurgitant flow rate and volume in patients with mitral regurgitation by Doppler color mapping of accelerating flow field. AB - OBJECTIVES: This study was designed to examine the accuracy of proximal accelerating flow calculations in estimating regurgitant flow rate or volume in patients with different types of mitral valve disease. BACKGROUND: Flow acceleration proximal to a regurgitant orifice, observed with Doppler color flow mapping, is constituted by isovelocity surfaces centered at the orifice. By conservation of mass, the flow rate through each isovelocity surface equals the flow rate through the regurgitant orifice. METHODS: Forty-six adults with mitral regurgitation of angiographic grades I to IV were studied. The proximal accelerating flow rate (Q) was calculated by: Q = 2 pi r2.Vn, where pi r2 is the area of the hemisphere and Vn is the Nyquist velocity. Radius of the hemisphere (r) was measured from two-dimensional or M-mode Doppler color recording. From the M-mode color study, integration of accelerating flow rate throughout systole yielded stroke accelerating flow volume and mean flow rate. Mitral regurgitant flow rate and stroke regurgitant volume were measured by using a combination of pulsed wave Doppler and two-dimensional echocardiographic measurements of aortic forward flow and mitral inflow. RESULTS: The proximal accelerating flow region was observed in 42 of 46 patients. Maximal accelerating flow measured from either two-dimensional (372 +/- 389 ml/s) or M-mode (406 +/- 421 ml/s) Doppler color study tended to overestimate the mean regurgitant flow rate (306 +/- 253 ml/s, p < 0.05). Mean Doppler accelerating flow rate correlated well with mean regurgitant flow rate (r = 0.95, p < 0.001), although there was a tendency toward slight overestimation of mean regurgitant flow by mean accelerating flow in severe mitral regurgitation. However, there was no significant difference between the mean accelerating flow rate (318 +/- 304 ml/s) and the mean regurgitant flow rate (306 +/- 253 ml/s, p = NS) for all patients. A similar relation was found between accelerating flow stroke volume (78.27 +/- 62.72 ml) and regurgitant flow stroke volume (76.06 +/- 59.76 ml) (r = 0.95, p < 0.001). The etiology of mitral regurgitation did not appear to affect the relation between accelerating flow and regurgitant flow. CONCLUSIONS: Proximal accelerating flow rate calculated by the hemispheric model of the isovelocity surface was applicable and accurate in most patients with mitral regurgitation of a variety of causes. There was slight overestimation of regurgitant flow rate by accelerating flow rate when the regurgitant lesion was more severe. PMID- 8426002 TI - Determinants of stroke volume response to exercise in patients with mitral stenosis: a Doppler echocardiographic study. AB - OBJECTIVES: The aim of this study was to assess exercise-induced changes in stroke volume and their main determinants in mitral stenosis. BACKGROUND: The mechanisms of the stroke volume response to exercise in mitral stenosis are not clearly established. METHODS: Twenty-seven patients with mitral stenosis, aged 47 +/- 13 years, and 10 healthy control subjects, aged 46 +/- 11 years, were examined by Doppler echocardiography to obtain stroke volume, mitral velocity time integral and calculated mitral valve area (by continuity equation) at rest and during submaximal supine bicycle exercise. Measured mitral valve area at rest and total mitral score were also obtained. RESULTS: During exercise, stroke volume increased significantly (p < 0.001) in the control subjects (+25 +/- 6%) but remained unchanged in the patients. In 10 patients (Group I), stroke volume increased by > or = 14% (+23 +/- 10%, p < 0.001); in the other 17 (Group II), it decreased or increased by < 14% (-5 +/- 14%, p = NS). Mitral velocity-time integral did not change in the three groups, whereas calculated mitral valve area increased significantly (p < 0.001) and similarly in Group I and the control group but remained unchanged in Group II. The exercise change in calculated mitral valve area correlated significantly with both measured mitral valve area at rest (r = 0.46, p < 0.05) and total mitral score (r - 0.53, p < 0.005). However, at constant mitral score, exercise change in calculated mitral valve area no longer correlated significantly with measured mitral valve area at rest. CONCLUSIONS: In mitral stenosis, the change in stroke volume during exercise depends on the change in mitral valve area, which itself depends on the degree of mitral valve damage. PMID- 8426003 TI - Percutaneous balloon mitral valvotomy with the Inoue single-balloon catheter: commissural morphology as a determinant of outcome. AB - OBJECTIVES: The aim of this study was to determine the importance to outcome and the predictability of commissural splitting in patients undergoing percutaneous mitral valvotomy with the Inoue single-balloon catheter. BACKGROUND: Echocardiographic scoring systems devised to predict mitral valvotomy outcome are based on assessment of leaflet and subvalvular morphology, but the specific importance of commissural morphology has not been examined. METHODS: In 30 consecutive patients, commissural splitting was predicted on the basis of the two dimensional echocardiographic commissural morphology: the extent of fusion, fibrosis or calcification of each commissure. Valve morphology also was evaluated according to a previously described echocardiographic scoring system. RESULTS: Splitting of one or both commissures occurred in 24 patients (80%) and was associated with a significantly greater mean increase in valve area (85%) than if neither commissure was split (13%). A good outcome from valvotomy (defined as valve area > 1.5 cm2 and increase in valve area > 25%) was achieved in 96% of those in whom one or both commissures split, but in none of the patients in whom neither commissure split. Whether or not splitting of at least one commissure would occur was predicted accurately in 28 (93%) of the 30 patients. Consequently, the prediction that one or both commissures would split was associated with a good outcome in 23 (89%) of 26 patients, whereas the prediction that neither commissure would split was not associated with a good outcome in any patient. There was no significant difference in the increase in mitral valve area between those with a mitral echocardiographic score < 8 and those with a score > or = 8. New or worsening mitral regurgitation occurred in nine patients, most commonly as a jet directed through a split commissure. CONCLUSIONS: Commissural splitting is the dominant mechanism by which mitral valve area is increased with the Inoue balloon technique, and it can be predicted by echocardiographic assessment of commissural morphology. Commissural morphology is a better predictor of outcome than is the mitral echocardiographic score. PMID- 8426004 TI - Echocardiographic description of the CarboMedics bileaflet prosthetic heart valve. AB - OBJECTIVES: The aim of this study was to describe the echocardiographic appearance of the normal CarboMedics prosthesis in the aortic and mitral positions. BACKGROUND: Echocardiography is the standard method of assessing prosthetic valves. However, new valve designs may still be marketed without an accompanying echocardiographic description. The CarboMedics prosthesis is in widespread use, but few noninvasive hemodynamic data have been published. METHODS: Echocardiography was performed in 147 patients with a total of 96 normally functioning CarboMedics prostheses in the aortic position and 75 in the mitral position; in 24 patients, valves were implanted in both positions. The following variables were measured: peak and mean transvalvular velocities, peak and mean instantaneous gradient estimated from the modified Bernoulli equation, aortic acceleration slope, pressure half-time, transvalvular flow and effective orifice area using the continuity equation. Patterns of regurgitation were observed by transthoracic study in all valves and by transesophageal study in selected mitral valve prostheses. RESULTS: For the aortic valve prostheses, estimated mean gradient ranged between 6 and 19 mm Hg. Effective area differed markedly among the anulus diameters (p < 0.001), with a mean value of 1 cm2 for the 19-mm valve and 2.6 cm2 for the 29-mm valve. For the mitral valve prostheses, mean gradient ranged from 3 to 7 mm Hg. There were a total of four washing leaks, one on either side of each pivotal point, and these lasted throughout systole or diastole. One jet was commonly more prominent than the other three. CONCLUSIONS: The CarboMedics prosthesis offered relatively little resistance to forward flow except at small anulus diameters. The washing jets were prominent and would be easy to misdiagnose as a sign of paraprosthetic regurgitation. PMID- 8426005 TI - Pulmonary artery hemodynamics in primary pulmonary hypertension. AB - OBJECTIVES: The present investigation compared and contrasted steady and pulsatile pulmonary hemodynamics at rest and during exercise in patients with primary pulmonary hypertension and normal control subjects. BACKGROUND: A complete description of the relation between pressure and flow in the pulmonary circulation includes both steady and pulsatile hemodynamic behavior. Patients with primary pulmonary hypertension provide a unique opportunity to study the effects of primary alterations in pulmonary vasculature on pulmonary artery vascular hydraulic load. METHODS: Catheter tip pressure and velocity recordings from the main pulmonary artery in 8 patients with primary pulmonary hypertension and 10 control subjects were used to derive the pulmonary artery input impedance spectrum and the extent of pulse wave reflection at rest and during exercise. RESULTS: As expected, in patients with primary pulmonary hypertension, mean pulmonary artery pressure (50 +/- 10 mm Hg) and pulmonary vascular resistance (880 +/- 446 dynes.s.cm-5) were markedly elevated at rest and remained so during exercise (mean pressure 71 +/- 15 mm Hg, mean resistance 750 +/- 530 dynes.s.cm 5). Pulmonary artery characteristic impedance was elevated at rest and did not change with exercise (rest 55 +/- 25 dynes.s.cm-5; exercise 66 +/- 33 dynes.s.cm 5). Measures of arterial wave reflection indicated that the extent of wave reflection in the pulmonary bed in those with primary pulmonary hypertension is large at rest (reflection coefficient 0.89 +/- 0.09) and that the composite reflected wave arrived during the midportion of right ventricular ejection. Although the extent of wave reflection decreased with exercise (reflection coefficient 0.81 +/- 0.10, p < 0.05), the magnitude and timing of these reflections remained adverse. Furthermore, in patients with primary pulmonary hypertension, the stroke volume response to exercise was strongly related to rest levels of pulmonary artery diastolic pressure, pulmonary vascular resistance and the reflection factor, whereas no such relation was found in the control subjects. CONCLUSIONS: In addition to the expected abnormalities in steady measures of pulmonary artery hemodynamics at rest in patients with primary pulmonary hypertension, rest and exercise measures of oscillatory behavior (characteristic impedance and pulse wave reflection) are perturbed. Measures of steady and pulsatile behavior, particularly wave reflection, appear to have an important role in the exercise response of these patients. PMID- 8426006 TI - Effects of adenosine in combination with calcium channel blockers in patients with primary pulmonary hypertension. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of vasodilator combination therapy in patients with primary pulmonary hypertension. BACKGROUND: Calcium channel blockers and adenosine have each been shown to be effective in reducing pulmonary artery pressure and pulmonary vascular resistance in patients with primary pulmonary hypertension. However, the effects of combining these vasodilators have not been studied. METHODS: To test the combination, 12 patients were placed on oral nifedipine and 3 on diltiazem therapy, using a dose titrated to maximal effect (mean nifedipine dose 103 +/- 24 mg, mean diltiazem dose 300 +/ 49 mg). Patients were then given maintenance doses of the calcium channel blocker at half the cumulative loading dose at 6-h intervals. One hour after the maintenance dose of calcium blocker, all patients received an infusion of adenosine, starting with 50 micrograms/kg per min and increasing by 50 micrograms/kg per min at 2-min intervals to a maximally tolerated dose (180 +/- 63 micrograms/kg per min). RESULTS: Ten patients responded to calcium channel blockers (defined as a > or = 20% decrease in pulmonary vascular resistance), with a 16% decrease in mean pulmonary artery pressure (p = 0.057), a 39% decrease in pulmonary vascular resistance (p = 0.002) and a 24% increase in stroke volume (p = 0.007). Five patients were nonresponders, with no significant changes in pulmonary artery pressure, pulmonary vascular resistance, cardiac index or stroke volume. In the calcium channel blocker responders, the combination of adenosine and calcium blocker reduced pulmonary vascular resistance by 49%, increased stroke volume by 33% and decreased mean pulmonary artery pressure by 14% compared with drug-free baseline values. In nonresponders, combination therapy resulted in nonsignificant changes in pulmonary artery pressure and pulmonary vascular resistance. CONCLUSIONS: Adenosine has the ability to further decrease pulmonary artery pressure and pulmonary vascular resistance in patients with primary pulmonary hypertension who respond to calcium channel blockers. Those who fail to respond to these agents have little added effect from adenosine. PMID- 8426007 TI - Hemodynamic correlates of the third heart sound during the evolution of chronic heart failure. AB - OBJECTIVES: The purpose of this study was to examine the temporal relation between the development of a third heart sound during the course of evolving heart failure and associated hemodynamic abnormalities. BACKGROUND: Although various theories have been proposed to explain the origin of the third heart sound, the exact origin of this sound remains unknown. METHODS: Studies were performed in seven dogs in which heart failure was produced by multiple sequential intracoronary micro-embolizations. Hemodynamic studies including ventriculography, pulsed wave Doppler echocardiography and intracardiac phonocardiography were performed at baseline, at the time at third heart sound was first heard and at 6 and 24 weeks after onset of the third heart sound. RESULTS: All dogs developed a third heart sound at 9 +/- 2 weeks after the initial embolization. The onset of the sound was accompanied by an increase in left ventricular chamber stiffness relative to the baseline value (0.25 +/- 0.03 vs. 0.14 +/- 0.01 mm Hg/ml) (p < 0.05) and mean deceleration of early mitral inflow velocity (1,040 +/- 90 vs. 590 +/- 40 cm/s per s) (p < 0.05). CONCLUSIONS: These data indicate that the onset of a third heart sound during the course of evolving heart failure occurs coincident with the development of increased left ventricular chamber stiffness and the manifestation of rapid deceleration of early mitral inflow velocity. These findings are consistent with a myocardial vibratory origin of this sound. PMID- 8426008 TI - Isolated ultrafiltration in moderate congestive heart failure. AB - OBJECTIVES: The aim of this study was to evaluate whether ultrafiltration is beneficial in patients with moderate congestive heart failure. BACKGROUND: Ultrafiltration is beneficial in patients with severe congestive heart failure. METHODS: We studied 36 patients in New York Heart Association functional classes II and III in stable clinical condition. Eighteen patients (group A) were randomly selected and underwent a single session of ultrafiltration (venovenous bypass, mean [+/- SEM] ultrafiltrate 1,880 +/- 174 ml, approximately 600 ml/h) and 18 (group B) served as control subjects. RESULTS: Two patients in group A and three in group B did not complete the 6-month follow-up study. In group A, soon after ultrafiltration there were significant reductions in right atrial pressure (from 8 +/- 1 to 3.4 +/- 0.7 mm Hg, pulmonary wedge pressure (from 18 +/- 2.5 to 10 +/- 1.9 mm Hg) and cardiac index (from 2.8 +/- 0.2 to 2.3 +/- 0.2 liters/min). During the follow-up period, lung function improved, extravascular lung water (X ray score) decreased and peak oxygen consumption (ml/min per kg) increased significantly from 15.5 +/- 1 (day -1) to 17.6 +/- 0.9 (day 4), to 17.8 +/- 0.9 (day 30), to 18.9 +/- 1 (day 90) and to 19.1 +/- 1 (day 180). Oxygen consumption at anaerobic threshold (ml/min per kg) also increased significantly from 11.6 +/- 0.8 (day -1) to 13 +/- 0.7 (day 4), to 13.7 +/- 0.5 (day 30), to 15.5 +/- 0.8 (day 90) and to 15.2 +/- 0.8 (day 180). These changes were associated with increased ventilation, tidal volume and dead space/tidal volume ratio at peak exercise. The improvement in exercise performance was associated with a decrease in norepinephrine at rest, a downward shift of norepinephrine kinetics at submaximal exercise and an increase in norepinephrine during orthostatic tilt. None of these changes were recorded in group B. CONCLUSIONS: In patients with moderate congestive heart failure, ultrafiltration reduces the severity of the syndrome. PMID- 8426009 TI - Radiofrequency ablation for atrioventricular node reentrant tachycardia: comparison between fast (anterior) and slow (posterior) pathway ablation. AB - OBJECTIVES: We compared the electrophysiologic effects on atrioventricular (AV) node physiology of selective "fast" versus selective "slow" pathway radiofrequency ablation in 42 patients with drug-resistant AV node reentrant tachycardia who underwent 51 ablation attempts to prevent tachycardia recurrence while preserving AV conduction. BACKGROUND: The recent introduction of radiofrequency ablation to treat AV node reentrant tachycardia allows the opportunity to study the effects of selective elimination of the different limbs involved in AV node reentrant tachycardia. METHODS: Selective fast pathway ablation was attempted in 13 patients by delivering radiofrequency energy anteriorly across the tricuspid valve anulus. Selective slow pathway ablation was attempted in 29 patients by delivering radiofrequency energy posteriorly across the tricuspid valve anulus at sites where putative slow pathway potentials were recorded. RESULTS: Selective fast pathway ablation eliminated AV node reentrant tachycardia without AV block in 6 (46%) of 13 patients after one ablation session and in an additional 3 patients (69% of total) after repeat ablation sessions. Slow pathway ablation eliminated AV node reentrant tachycardia without AV block in 26 (90%) of 29 patients after one radiofrequency ablation session and in an additional 2 patients (97% of total) after repeat ablation sessions. Selective fast pathway ablation increased the PR interval (140 to 220 ms, p = 0.0001) and AH interval (66 to 153 ms, p = 0.0001), whereas slow pathway ablation did not change these intervals. Fast pathway radiofrequency ablation caused retrograde block in 7 (64%) of 11 patients, whereas no patients undergoing slow pathway ablation developed selective retrograde block. Single AV node echo beats were commonly induced after slow but not fast pathway ablation (17 of 29 patients vs. 1 of 11 patients, respectively, p = 0.01) and did not predict recurrence of AV node reentrant tachycardia. CONCLUSIONS: Successful selective radiofrequency ablation of fast or slow pathways in patients with AV node reentrant tachycardia resulted in different electrophysiologic properties after ablation. Slow pathway ablation produced more successful outcomes, with a decreased prevalence of recurrent AV node reentrant tachycardia or AV block. PMID- 8426010 TI - Hematologic correlates of left atrial spontaneous echo contrast and thromboembolism in nonvalvular atrial fibrillation. AB - OBJECTIVES: This study examined the relation between left atrial spontaneous echo contrast, hematologic variables and thrombo-embolism in patients with nonvalvular atrial fibrillation. BACKGROUND: Left atrial spontaneous echo contrast is associated with left atrial stasis and thromboembolism in patients with nonvalvular atrial fibrillation. However, its hematologic determinants in patients with nonvalvular atrial fibrillation are unknown. METHODS: Clinical, hematologic and echocardiographic variables were prospectively measured in 135 consecutive patients with nonvalvular atrial fibrillation undergoing transesophageal echocardiography. RESULTS: Patients with left atrial spontaneous echo contrast (n = 74, 55%) had an increased fibrinogen concentration (p = 0.029), platelet count (p = 0.045), hematocrit (p = NS) and left atrial dimension (p = 0.005). Multivariate analysis showed that left atrial spontaneous echo contrast was independently related to hematocrit (odds ratio = 2.24, p = 0.002), fibrinogen concentration (odds ratio = 2.08, p = 0.008) and left atrial dimension (odds ratio = 1.90, p = 0.004) but not platelet count. It was also associated with left atrial thrombus (n = 15, p = 0.001) and with recent embolism (n = 40, p < 0.001). In 40 clinically stable outpatients without previous embolism, left atrial spontaneous echo contrast was significantly related to hematocrit (p = 0.005), fibrinogen concentration (p = 0.035) and left atrial dimension (p = 0.029) but not to coagulation factor VII, D-dimer, erythrocyte sedimentation rate, platelet count, plasma beta-thromboglobulin, plasma glycocalicin or glycocalicin index. CONCLUSIONS: Left atrial spontaneous echo contrast in patients with nonvalvular atrial fibrillation is independently related to hematocrit, fibrinogen concentration and left atrial dimension, indicating a relatively hypercoagulable state in addition to stasis. These findings support the hypothesis that left atrial spontaneous echo contrast is due to erythrocyte aggregation. Hematologic factors may contribute to its association with thromboembolism. PMID- 8426011 TI - Selective anterograde coronary arteriography in neonates with d-transposition of the great arteries: accuracy and safety. AB - OBJECTIVES: This study was designed to evaluate the accuracy and safety of selective anterograde coronary arteriography for the identification of the origin and branching pattern of the three main coronary arteries in neonates with d transposition of the great arteries. BACKGROUND: Definition of coronary artery anatomy is important in neonates with d-transposition of the great arteries who are considered for the arterial switch operation. Balloon occlusion aortography defines coronary artery anatomy in most but not all cases. We have described a technique for selective anterograde coronary arteriography. METHODS: Between March 1987 and May 1991, 17 neonates underwent selective anterograde coronary arteriography and 29 patients had balloon occlusion aortography. After venous access was gained, a mesenteric catheter was used to engage the coronary ostia for contrast injection. All angiograms were reviewed by three independent observers and the coronary artery diagnoses were compared with operative findings. Complications with the catheterization procedure were also recorded. RESULTS: The accuracy of defining coronary artery anatomy with selective anterograde coronary arteriography (98 +/- 2%) was significantly (p < 0.05) greater than that achieved with balloon occlusion aortography (69 +/- 6%). There were no deaths with catheterization in either study group. Morbidity was similar in the groups with balloon occlusion aortography (7%) and selective anterograde coronary arteriography (6%) (p > 0.05) and was related to transient bradycardia induced by catheter manipulation in the right ventricle. No patient in either study group experienced cardiac ischemia. CONCLUSIONS: Selective anterograde coronary arteriography is an accurate and safe technique for the definition of coronary artery pattern in neonates with d-transposition of the great arteries. PMID- 8426012 TI - Enlarged bronchial arteries after early repair of transposition of the great arteries. AB - OBJECTIVES: This study was undertaken to define the incidence of enlarged bronchial arteries after early surgical repair of transposition of the great arteries by the arterial switch operation, and to report the results of catheter directed therapy in five patients. BACKGROUND: Pathologic and angiographic studies have demonstrated enlarged bronchial arteries in patients with transposition of the great arteries. METHODS: A subjective 4-point scale was used to grade postoperative angiograms performed in 119 patients at our institution between January 1983 and December 1991. Grades 0 and 1 were designated if there was no opacification of the pulmonary arteries or veins, whereas grades 2 and 3 were assigned if there was such opacification. The median age at repair was 8 days (range 1 day to 13 months) and the median age at catheterization was 11.2 months (range 3.6 to 58.5). An intact ventricular septum was present in 84 (71%) of 119 patients. RESULTS: Significantly increased bronchial flow (grade 2 or 3) was present in 55 (46%) of 119 patients. Age at repair, age at catheterization and interval between repair and catheterization were not associated with significantly increased bronchial flow; however, an intact ventricular septum was weakly associated with increased flow (p = 0.04). Coil embolization was performed in five patients with complete occlusion of the vessels and no significant complications. CONCLUSIONS: Abnormally enlarged bronchial arteries are frequently identified at postoperative catheterization despite early repair and may explain continuous murmurs or persistent cardiomegaly in patients with otherwise normal noninvasive findings. When clinically indicated, catheter-directed therapy can be performed with good results. PMID- 8426013 TI - Effect of zatebradine on contractility, relaxation and coronary blood flow. AB - OBJECTIVES: The purpose of this study was to compare the effects of zatebradine on heart rate, contractility and relaxation with those of its structural analog verapamil. We used isoproterenol, a potent beta-agonist, to see how these effects were modulated by sympathetic activation. We also compared the effects of zatebradine and verapamil on coronary blood flow and coronary blood flow reserve. BACKGROUND: Zatebradine, previously called UL-FS 49, is a new bradycardic agent believed to act selectively at the sinoatrial node. METHODS: Isolated isovolumetric pig hearts were prepared and left ventricular pressure, its first derivative (dP/dt), tau and heart rate were measured both before and after administration of either 0.975 mg of zatebradine (Group I, n = 8) or 125 micrograms of verapamil (Group II, n = 8). After the effects of each drug reached a plateau, a continuous infusion of isoproterenol was started and measurements were obtained again and compared with a third group of measurements from control hearts infused with isoproterenol after receiving only saline solution (n = 8). We also assessed the effects of zatebradine and verapamil on coronary vascular tone by measuring flow in the left anterior descending coronary artery in intact anesthetized open chest pigs both before and after the intracoronary administration of these drugs (n = 8 for each). All preparations were atrially paced to negate any bradycardiac effects of the drugs. RESULTS: In the group that received zatebradine, mean (+/- SE) heart rate decreased from 143 +/- 8 to 99 +/- 4 beats/min (p < 0.01) and there was no significant change in either peak left ventricular systolic pressure, dP/dt or tau. In contrast, verapamil produced a lesser decrease in heart rate (136 +/- 7 to 120 +/- 7 beats/min, p < 0.05) but produced substantial decreases in peak left ventricular pressure (100 +/- 3 to 45 +/- 4 mm Hg, p < 0.01) and dP/dt (68% decrease, p < 0.01) and an increase in tau (+26%, p < 0.05). Isoproterenol restored these variables toward normal values in the hearts treated with verapamil, although left ventricular systolic pressure and dP/dt were restored to control values only at the highest isoproterenol concentrations. In the hearts treated with zatebradine, isoproterenol significantly increased left ventricular pressure and contractility and decreased tau; however, heart rate remained unchanged at peak effect. Zatebradine had no effect on coronary blood flow and there was a 100% increase in flow with reactive hyperemia. Conversely, verapamil increased coronary flow by 100%, with no subsequent further increase by reactive hyperemia compared with control values. CONCLUSIONS: Although structurally similar to verapamil, zatebradine is a highly specific bradycardic agent. It has little direct effect on left ventricular developed pressure, contractility, relaxation and coronary vascular tone. Furthermore, the bradycardic effect of zatebradine unlike that of verapamil, is not overcome by doses of isoproterenol that increase developed pressure and contractility and improve relaxation. Because of its highly specific bradycardic effect, this drug may potentially be useful in treating patients with ischemic heart disease or congestive heart failure. PMID- 8426014 TI - Doppler color flow mapping of epicardial coronary arteries: initial observations. AB - OBJECTIVES: We addressed the hypothesis that blood flow could be imaged by Doppler color flow mapping of the coronary arteries and characteristic patterns described in normal and diseased vessels. BACKGROUND: Echocardiographic imaging of the epicardial coronary arteries has been suggested as a useful adjunct to their intraoperative evaluation. Addition of Doppler color flow mapping could potentially enhance this evaluation by displaying the flow disturbance produced by anatomic lesions whose physiologic significance may otherwise be uncertain. In experimental models, such displays could also potentially provide insights into the pathophysiology of coronary blood flow and stenosis. METHODS: Epicardial coronary arteries were examined with a high resolution 7-MHz linear phased-array transducer both in vivo and in vitro. 1) The coronary arteries were studied in the beating hearts of 10 open chest dogs in which experimental stenoses were also created; the maximal extent of the arterial tree in which flow could be seen in the most ideal setting was also examined in four additional excised perfused canine hearts. 2) Six excised human coronary arteries were perfused in a pulsatile manner to determine whether abnormal flow patterns could be prospectively identified and subsequently correlated with pathologic evidence of stenosis. RESULTS: All normal coronary artery segments studied showed homogeneous flow without evidence of flow disturbance. In the excised heart, flow could be visualized to the distal extent of the epicardial vessels; in the open chest model, visualization of the proximal 5 to 6 cm was comparable, although surrounding structures limited access to the terminal portions of the vessels. The stenotic lesions created in the canine hearts (n = 9) showed recognizable alterations in the flow pattern: localized aliasing, proximal blood flow acceleration, distal flow disturbance and recirculatory flow. In the excised human arteries, these features identified 12 lesions, all of which corresponded to areas of > or = 50% lumen narrowing by pathologic examination. CONCLUSION: Blood flow in the epicardial coronary arteries can be imaged by Doppler color flow mapping and characteristic flow patterns described in normal and diseased vessels. PMID- 8426015 TI - Exogenous prostacyclin decreases vasoconstriction but not platelet thrombus deposition after arterial injury. AB - OBJECTIVES: The aim of this study was to examine the in vivo effects of increasing doses of prostacyclin (PGI2) on arterial vasoconstriction, platelet deposition and their interrelation after balloon injury of porcine carotid arteries. BACKGROUND: Extensive platelet deposition and localized vasoconstriction occur acutely after arterial injury in vivo. The platelet deposition and vasoconstriction are directly correlated, and previous studies suggest that platelets may mediate the vasoconstrictive response. However, it is unclear whether vasoconstriction contributes to platelet deposition. METHODS: Seven pigs received an intravenous infusion of PGI2 at 10 ng/kg per min (PGI2 10), 8 pigs at 50 ng/kg per min (PGI2 50) and 4 pigs at 500 ng/kg per min (PGI2 500); 24 pigs with saline infusion served as a control group. RESULTS: Vasoconstriction immediately proximal and distal to the balloon-dilated carotid arterial segment where selective endothelial injury occurred was directly related to indium-111-labeled platelet deposition within the dilated segment in both control pigs and PGI2-treated pigs. However, this relation was such that for any given level of platelet deposition relative to control, PGI2 decreased vasoconstriction in a dose-related manner. None of the treatments (PGI2 10, 50 or 500) decreased quantitative 111In-labeled platelet deposition or the proportion of deeply injured arteries with mural thrombus (91%, 70% or 75%, respectively, p = NS) compared with values in control pigs (81%). Thus, vasoconstriction was directly related to platelet deposition in control and PGI2-treated animals, but vasodilation alone did not decrease platelet deposition. CONCLUSIONS: Intravenous infusion of PGI2 significantly decreases vasoconstriction but not platelet deposition or mural thrombosis after arterial injury by balloon dilation. It is therefore unlikely that vasoconstriction mediates platelet deposition in this model. At hemodynamically tolerated doses, PGI2 infusion probably will not prevent the thrombotic complications associated with angioplasty. PMID- 8426016 TI - Endogenous prostaglandin endoperoxides may alter infarct size in the presence of thromboxane synthase inhibition: studies in a rabbit model of coronary artery occlusion-reperfusion. AB - OBJECTIVES: The aim of this study was to assess whether prostaglandin endoperoxides, which continue to be formed in the setting of thromboxane A2 synthase inhibition, might influence the fate of ischemic myocardium in a model of coronary occlusion and reperfusion. BACKGROUND: It was recently demonstrated that thromboxane A2 synthase inhibitors reduce ischemic myocardial injury through a redirection of prostaglandin (PG) endoperoxides toward the synthesis of "cardioprotective" prostaglandins, such as PGI2, PGE2 and PGD2. However, part of these prostaglandin endoperoxides may also stimulate a receptor, shared with thromboxane A2, mediating platelet aggregation and vasoconstriction. METHODS: New Zealand White rabbits were subjected to 30 min of coronary occlusion, followed by 5.5 h of reperfusion. Fifteen minutes before reperfusion, the animals were randomized to receive 1) saline solution (control animals, n = 8); 2) SQ 29548, a potent and selective thromboxane A2/PGH2 receptor antagonist (n = 8); 3) dazoxiben, a selective thromboxane A2 synthase inhibitor (n = 8); 4) R 68070 (Ridogrel), a drug with dual thromboxane A2 synthase-inhibiting and thromboxane A2/PGH2 receptor-blocking properties (n = 8); or 5) aspirin + R 68070 (n = 8). RESULTS: Dazoxiben and R 68070, but not SQ 29548, significantly reduced thromboxane B2 formation and increased plasma levels of 6-keto-PGF1 alpha, PGE2 and PGF2 alpha. Ex vivo platelet aggregation induced by U46619 (a thromboxane A2 mimetic) was inhibited by SQ 29548 and R 68070 but not by dazoxiben. In control animals, infarct size determined at the end of the experiment by triphenyltetrazolium chloride staining averaged 57.7 +/- 3.2% of the area at risk of infarction. The administration of SQ 29548 did not significantly reduce infarct size compared with that in control animals, whereas dazoxiben and R 68070 significantly reduced infarct size to 36.7 +/- 2.8% and 16.6 +/- 3.6% of area at risk of infarction, respectively (p < 0.001 vs. control values). In rabbits treated with R 68070, infarct size was also significantly smaller than that of dazoxiben-treated rabbits (p < 0.01). This protective effect of R 68070 was completely abolished when the drug was administered with aspirin, infarct size in this group averaging 59.7 +/- 1.6% (p = NS vs. control values). No differences in regional myocardial blood flow, systemic blood pressure, heart rate or extent of area at risk were observed among groups. CONCLUSIONS: Thus, prostaglandin endoperoxides play an important role in modulating the cardioprotective effects of thromboxane A2 synthase inhibitors. The simultaneous inhibition of thromboxane A2 synthase and blockade of thromboxane A2/PGH2 receptors by R 68070 identify a pharmacologic interaction of potential therapeutic importance. PMID- 8426018 TI - Effects of aspirin in arterial thrombosis: why don't animals behave the way humans do? PMID- 8426017 TI - High dose intravenous aspirin, not low dose intravenous or oral aspirin, inhibits thrombus formation and stabilizes blood flow in experimental coronary vascular injury. AB - OBJECTIVES: The purpose of this study was to assess the anti-thrombotic potential of various forms of aspirin administration. BACKGROUND: Platelet activation in response to endothelial injury has been implicated in acute coronary syndromes. METHODS: Delivering 100-microA anodal direct current to the intima of the left circumflex coronary artery in dogs at a site of moderate external stenosis provides a thrombogenic model of vascular injury. Animals were treated with aspirin (Group I, 20 mg/kg intravenously [n = 11]; Group II, 4.6 mg/kg intravenously [n = 6]; Group III, 4.6 mg/kg orally 18 h before the experiment [n = 7]) or vehicle (Group IV, control [n = 11]). RESULTS: The time required for thrombotic occlusion to occur was longer and the incidence of thrombosis was lower in Group I (Group I, 238 +/- 7 min [n = 2]; Group II, 127 +/- 25 min [n = 3]; Group III, 156 +/- 35 min [n = 6]; Group IV, 90 +/- 11 min [n = 11]) (p < 0.05). Thrombus mass was smaller in Group I (Group I, 5.0 +/- 0.8 mg; Group II, 12.2 +/- 2.6 mg; Group III, 11.6 +/- 3.9 mg; Group IV, 9.1 +/- 1.6 mg) (p < 0.05). Initial hemodynamic variables did not differ among groups. An increase in mean arterial pressure was noted for several hours after intravenous aspirin administration in Group I (99 +/- 5 to 110 +/- 4 mm Hg) (p < 0.05). Left circumflex coronary artery blood flow was stable for 5 h in Group I (Group I, 31 +/- 2 to 26 +/- 4 ml/min) but decreased in all the other groups (Group II, 26 +/- 4 to 10 +/- 5 ml/min; Group III, 27 +/- 5 to 7 +/- 7 ml/min; Group IV, 29 +/- 4 to 0 ml/min) (p < or = 0.05). The in vivo area of left ventricle perfused by the left circumflex coronary artery was not different among groups. Platelet counts were similar and did not change over the course of the protocol. Ex vivo arachidonic acid-induced platelet aggregation decreased in all groups after aspirin (p < or = 0.001). Indium-111-labeled platelet adherence to the coronary vasculature was decreased in distal vessel segments after all doses of aspirin (p < 0.05). Platelet deposition in thrombi was similar for all treatment groups. CONCLUSIONS: High dose intravenous aspirin has salutary effects. It stabilizes left circumflex coronary artery blood flow, prolongs the time to thrombosis, reduces the incidence of thrombotic occlusion, reduces thrombus mass and limits platelet adherence to sites of arterial injury. Low dose aspirin given intravenously or orally was ineffective. When persistent intracoronary thrombi precipitate unstable coronary syndromes, high dose intravenous aspirin may be useful in the acute period even though platelets continue to interact with injured vascular segments through aspirin-insensitive mechanisms. PMID- 8426019 TI - Long-term oral nitrate therapy prevents chronic ventricular remodeling in the dog. AB - OBJECTIVES: The purpose of this experiment was to assess the long-term effects of oral nitrate therapy on ventricular remodeling in a canine model of discrete myocardial damage. BACKGROUND: A progressive increase in left ventricular mass and volume has been documented after experimental and clinical myocardial infarction. This ventricular remodeling has been associated with the development of congestive heart failure. Nitrate therapy, especially when combined with hydralazine, is effective in the management of clinical heart failure. Moreover, nitrates inhibit infarct expansion, one of the earliest manifestations of ventricular remodeling. Whether nitrates can attenuate chronic left ventricular remodeling is unknown. METHODS: Dogs with discrete myocardial necrosis produced 24 h earlier by transmyocardial direct current shock were randomized to receive isosorbide mononitrate, 30 mg twice daily (n = 10), or no treatment (n = 4); the latter group was augmented by 13 control dogs from a prior study in which an identical protocol was used. Ventricular structure was assessed with nuclear magnetic resonance imaging at baseline and at 1 and 16 weeks after myocardial damage. RESULTS: Left ventricular mass increased at 1 week in the control group (mean +/- SD 68.1 +/- 10.7 g to 80.1 +/- 12.1 g, p = 0.0001) but not in the nitrate-treated group (70.2 +/- 7.7 g to 69.6 +/- 7.3 g, p = NS). No change in left ventricular volume was observed in either group during the 1st week after myocardial damage. After 16 weeks of follow-up left ventricular mass had increased by 12.7 +/- 7.1 g (p = 0.001) in the control group and had decreased by 1.2 +/- 7.7 g in the nitrate group. Left ventricular volume was increased at 16 weeks by 14.2 +/- 10.3 ml in the control group but was decreased by 3.7 +/- 7.5 ml in the nitrate group. Isosorbide mononitrate produced transient hemodynamic effects with a return of most measured variables toward baseline within 2 h after administration of the drug. At 1 week there was no intergroup difference in rest hemodynamic variables assessed 12 h after drug administration. At 16 weeks, pulmonary capillary wedge pressure (15 +/- 4 vs. 8 +/- 3 mm Hg, p = 0.0001), pulmonary artery pressure (24 +/- 5 vs. 16 +/- 3 mm Hg, p = 0.0001) and right atrial pressure (10 +/- 3 vs. 6 +/- 3 mm Hg, p = 0.008) were all higher in the control group. CONCLUSIONS: Long-term oral nitrate therapy attenuates the early and late manifestations of ventricular remodeling after myocardial damage in the dog. Hemodynamic observations suggest the possibility that drug-induced preload or afterload reduction does not completely explain this effect. PMID- 8426020 TI - Selective elimination of retrograde conduction by intraoperative neodymium: YAG laser photocoagulation in dogs. AB - OBJECTIVES: The purpose of this study was to test the feasibility of selective elimination of ventriculoatrial (VA) conduction by limited laser photocoagulation of the atrioventricular (AV) node, and to analyze the histologic substrate of unidirectional retrograde block. BACKGROUND: Atrioventricular node reentry requires intact retrograde conduction. METHODS: Neodymium:yttrium-aluminum-garnet laser photocoagulation was performed during cardiopulmonary bypass through a right atriotomy in 15 dogs that had intact retrograde conduction before operation. Short laser pulses were delivered to an area between the coronary sinus orifice and the proximal His bundle. The end point of lasing was second degree AV node block at a paced atrial cycle length of 250 ms. RESULTS: Complete retrograde block developed immediately in 11 of the 15 dogs (group I), while AV conduction persisted in all 11. In 4 of the 15 dogs (group II), both AV and VA conduction remained intact. During a 3-month follow-up period, retrograde conduction remained absent in all group I dogs. Retrograde block was not reversed by isoproterenol. Anterograde AV node characteristics (Wenckebach cycle length, functional refractory period, ventricular rate during atrial fibrillation) were unchanged in five dogs and modified in six. Complete AV block did not develop. In four control dogs (group III, sham operation), anterograde and retrograde AV node characteristics were unchanged. The anterograde Wenckebach cycle lengths in groups I, II and III at 3 months measured 192 +/- 15 ms, 195 +/- 6 ms and 170 +/- 22 ms, respectively, whereas the retrograde Wenckebach cycle lengths in groups II and III measured 345 +/- 62 ms and 278 +/- 25 ms, respectively. Histologic study at 3 months in cases with unidirectional VA block showed the compact part of the AV node intact with destruction of the atrial approaches and the superficial layers of the proximal end of the node on the right side. CONCLUSIONS: 1) With limited laser photocoagulation of the proximal AV node area, VA conduction can be eliminated and anterograde AV node transmission maintained. 2) Destruction of the atrial approaches on the right side with preservation of the compact part of the AV node may result in unidirectional retrograde block. PMID- 8426021 TI - Cardiovascular effects of lightning strikes. AB - OBJECTIVES: The purpose of this study was to investigate the effects of lightning strikes on the cardiovascular system. BACKGROUND: A lightning strike can attack its victims in one of three ways: direct hit, splash or ground strike. The cardiovascular system can be affected directly by mechanical or electrical trauma during a direct hit or can be indirectly affected through effects on the total body with extensive catecholamine release or autonomic stimulation. Reported effects include hypertension, tachycardia, nonspecific electrocardiographic (ECG) changes including prolongation of the corrected QT (QTc) interval, transient T wave inversion and myocardial necrosis with creatine kinase-MB (CK-MB) fraction release. METHODS: Nineteen victims from five separate lightning strikes were studied over a 2-month period. Each patient was evaluated by serial ECG, CK-MB determinations and echocardiography. RESULTS: The early (0 to 72 h) effects of lightning were demonstrated on the ECG by ST segment elevation consistent with acute current of injury, prolonged QTc interval with direct hits and nonspecific ST and T wave changes. On echocardiography, segmental or global ventricular dysfunction was seen, and pericardial effusion was also detected. During the intermediate (3- to 14-day) period, new and often marked ECG changes consistent with pericarditis or ischemia were seen. No new echocardiographic changes were detected, however, and the early abnormalities including severe left ventricular dysfunction with cardiogenic shock have reversed. The late (1 to 12 months) period revealed only one patient with long-term sequelae (recurrent pericarditis that persisted for 5 months). CONCLUSIONS: Unless both entrance and exit sites are limited to the lower limbs, direct and splash lightning strikes cause myocardial damage as assessed by abnormal serum enzyme determinations or abnormal echocardiographic findings. Only direct hits resulted in echocardiographic abnormalities or a prolonged QTc interval. The degree of myocardial injury can be severe with left and right ventricular ejection fraction < 15% and can be reversible. PMID- 8426022 TI - Does reperfusion injury exist in humans? AB - Timely coronary reperfusion as treatment for acute myocardial infarction reduces myocardial infarct size, improves left ventricular function and survival. There is still concern that at the time of reperfusion, a further injury occurs to the myocardium. Theoretically, if this "reperfusion injury" could be treated and eliminated, the outcome for patients with myocardial infarction might further improve. The concept of reperfusion injury is closely tied to the concept that oxygen radicals generated at the time of reperfusion cause tissue damage. There are four basic forms of reperfusion injury. Lethal reperfusion injury is described as myocyte cell death due to reperfusion itself rather than to the preceding ischemia. This concept continues to be controversial in both experimental animal and clinical studies. Vascular reperfusion injury refers to progressive damage to the vasculature over time during the phase of reperfusion. Manifestations of vascular reperfusion injury include an expanding zone of no reflow and a deterioration of coronary flow reserve. This form of reperfusion injury has been documented in animal models and probably occurs in humans. Stunned myocardium refers to postischemic ventricular dysfunction of viable myocytes and probably represents a form of "functional reperfusion injury." This phenomenon is well documented in both animal models and humans. Reperfusion arrhythmias represent the fourth form of reperfusion injury. They include ventricular tachycardia and fibrillation that occur within seconds to minutes of restoration of coronary flow after brief (5 to 15 min) episodes of myocardial ischemia. True reperfusion arrhythmias occur in only a small percentage of patients receiving thrombolytic therapy for acute myocardial infarction and are not a sensitive indicator for successful reperfusion.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426023 TI - Recommendations for peripheral transluminal angioplasty: training and facilities. American College of Cardiology Peripheral Vascular Disease Committee. PMID- 8426024 TI - Do you want your child to become a doctor? PMID- 8426025 TI - Relation of peak atrial filling velocity and end-diastolic stiffness: fact or fancy? PMID- 8426026 TI - Increased left ventricular outflow tract obstruction during exercise in patients with hypertrophic cardiomyopathy. PMID- 8426027 TI - Subgroup analyses in the I.S.A.M. trial. PMID- 8426028 TI - Inpatient community-based geriatric assessment reduces subsequent mortality. AB - OBJECTIVE: To evaluate the effect of an Inpatient Geriatric Consultation Team on patient outcome. DESIGN: Randomized controlled clinical trial. SETTING: A non academic-affiliated 503-bed community hospital. PATIENTS: All inpatients over the age of 70 years. Sixty-two patients received multidimensional geriatric assessment, and 58 patients received no intervention. INTERVENTION: Team assessment, leading to formal recommendations to the attending physician. MEASUREMENTS: Data were collected on hospital length of stay, referrals to community service, discharge destination, hospital readmissions in 6 months, number of post-discharge physician visits, and change in functional status. Mortality at 6 months and at 1 year was determined for each patient. MAIN RESULTS: At 6 months, 12/58 patients (21%) had died in the control group versus 3/62 (6%) patients in the experimental group (P = 0.01). During hospitalization, the length-of-stay was 10.1 days for the control group versus 9.0 days for the experimental group (P = 0.20). The control group had significantly more readmissions (0.6 per patient vs 0.3 per patient, P = 0.02). A higher number of experimental patients, 22% (13/59), showed improvement in ADL scores compared with 7% (4/46) of control patients, P = 0.07. At one year for all randomized patients, 7/68 (10%) of experimental patients and 13/64 (20%) of control patients had died. CONCLUSIONS: Short-term mortality can be reduced in community inpatient acute hospital settings by comprehensive geriatric consultation teams. Important differences in mortality remain at 1 year of followup. Trends towards improved functional status and fewer hospital readmissions favor the intervention group. PMID- 8426029 TI - Adverse clinical events in dependent long-term nursing home residents. AB - OBJECTIVE: To assess the occurrence, type, and burden of adverse clinical events (ACE) among residents of chronic facilities. An ACE is any acute or subacute change in health status suggesting acute or subacute illness. DESIGN: Survey with face-to-face functional assessment and 3-month retrospective chart review. STUDY POPULATION: One hundred six continuing-care residents with a minimum length of stay of 4 months at an Italian chronic care facility. MEASUREMENTS: Functional assessment and chart review-based classification of ACE burden on care management according to a clinical-functional Severity Rating Scale. MAIN RESULTS: Functional dependence, dementia and concurrent clinical problems were common. Two hundred seventy-three ACEs were detected. Eighty-nine percent of residents experienced at least one ACE. Only 21% of ACEs could be managed by simple medical intervention and monitoring within 1 day; in 23% there was need for more complex care management; 7% of the latter ACEs resulted in residual (new) functional impairment. Cardiovascular and gastrointestinal systems were most commonly involved in ACEs. Neurological ACEs were the most frequent category leading to new functional impairment (22%). ACE occurrence/burden was higher in male residents (P < 0.01) and strongly associated with the number of concurrent medical problems (P < 0.001). Neither cognitive nor functional dependence levels were related to ACE occurrence. CONCLUSIONS: This description of the burden on care management resulting from acute and subacute changes in clinical and functional status of chronic patients emphasizes the continuing and unpredictable nature of medical attention required in a nursing home or chronic care facility. ACEs occur far more frequently among the elderly than is generally recognized. Thus a high level of medical and nursing skill is necessary in chronic care facilities. PMID- 8426030 TI - Public opinion regarding consent to treatment. AB - OBJECTIVE: To examine public opinion regarding certain elements of consent: disclosure, advance directives, substitute decisions, emergency treatment, and advocacy. DESIGN: Cross-sectional random-digit-dialing telephonic survey. SETTING: General public. PARTICIPANTS: One thousand randomly-selected adults living in Ontario. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Self-reported attitudes and behaviors. RESULTS: Of 5,708 telephone numbers drawn, 641 were non valid or non-residential, 1,283 were not in service, 1,068 had no answer, a busy line, or an unavailable respondent, 147 subjects had a language barrier, 1,497 subjects refused, 72 interviews were incomplete, and 1,000 interviews were completed. With regard to disclosure, 33% of respondents said that a doctor should withhold information from a patient if asked to do so by the patient's family. With regard to advance directives, 36% of respondents had had advance discussions with their families, and 12% had completed a living will. With regard to substitute decisions, 77% of respondents said that they would went their wishes followed if they were unable to make medical decisions for themselves; 58% wanted their spouse or partner to make such decisions for them. With regard to emergency treatment, 48% of respondents stated that a doctor should give a life saving blood transfusion to an unconscious adult carrying a card stating that blood transfusion was against his or her religious beliefs. With regard to advocacy, 78% of respondents supported mandatory advocacy services for serious health decisions such as whether to have heart surgery; 33% supported advocacy services for less serious decisions such as whether to have a dental filling. CONCLUSIONS: These data highlight the need to reconsider legislative provisions regarding and/or target public education programs toward specific consent-related issues including disclosure, advance directives, substitute decisions, emergency treatment, and advocacy. PMID- 8426031 TI - Observations on the prevalence of protein-calorie undernutrition in VA nursing homes. AB - OBJECTIVE: To determine the prevalence of underweight and hypoalbuminemia in Veterans Affairs nursing home residents and the frequency with which physicians, nurses, and dietitians documented their awareness of the presence of underweight and hypoalbuminemia. DESIGN: Retrospective survey of medical records. SETTING: Twenty-six Department of Veterans Affairs nursing homes located in the Department's Central Region. PATIENTS: 2811 residents. MAIN OUTCOME MEASURES: (1) Prevalence of underweight (defined as body weight less than 80% of standard) and hypoalbuminemia (defined as serum albumin less than 3.5 g/dl); (2) frequency with which physicians, nurses, and dietitians documented the prevalence of underweight and hypoalbuminemia in the records. RESULTS: Three hundred thirty-two (11.8%) of the patients were underweight and 772 (27.5%) were hypoalbuminemic. The prevalence of the two conditions varied widely across nursing homes. The prevalence of low serum albumin ranged from 5% to 58% in the 26 institutions, and the prevalence of body weight less than 80% of standard ranged from 2% to 20%. In the Region as a whole, the dietician's notes in the medical charts mentioned underweight in an average of 95% of affected cases but mentioned the suboptimal albumin level in only 82% of the hypoalbuminemic individuals. In the nurses's notes, these figures were only 80% and 45%, respectively, and in the physician's notes, only 62% and 46%, respectively. The frequency of documented awareness of underweight and hypoalbuminemia by nurses and physicians varied tremendously across the 26 facilities (as few as 7% of the undernourished cases in one nursing home, 100% of the affected individuals in another). CONCLUSIONS: This study showed a high prevalence of calorie and protein undernutrition in the nursing home residents of VA nursing homes, wide variation in the prevalence across nursing homes, and frequent lack of documentation of these nutritional deficiencies by physicians and nurses. PMID- 8426032 TI - Clinical presentation of meningioma in the elderly. AB - OBJECTIVE: To compare presenting manifestations of meningioma in elderly patients with those in younger patients. DESIGN: Retrospective survey. SETTING: Tertiary care center. PATIENTS: All hospitalized adult patients with the primary diagnosis of meningioma over a 5-year period (n = 116). Nine patients with recurrence of a previously resected meningioma were excluded. Fifty-two percent (56/107) of patients were > or = 65 years of age. MEASUREMENTS: Presenting symptoms were classified as asymptomatic, focal, non-specific, or both focal and non-specific. Presenting neurologic signs on physical examination were classified as focal, non focal, or normal. Duration of symptoms was classified as < or = 1 week, 1 week to 1 month, 1 to 6 months, and > 6 months. RESULTS: Older patients presented more commonly with confusion (32% vs 4%) and dystaxia (28% vs 10%) than younger patients and presented less commonly with headache (23% vs 49%) and visual changes (21% vs 43%). While 2% of patients were asymptomatic at presentation, the remainder had symptoms categorized as focal (38%), non-specific (26%), or both (34%). There was no significant difference between older and younger age groups in the proportions of patients having focal symptoms. Duration of symptoms for all patients was categorized as < or = 1 week (15%), 1 week to 1 month (16%), 1 to 6 months (23%), and > 6 months (44%). In general, older patients were diagnosed earlier than younger patients. The presence of focal neurologic signs was not significantly different between older and younger patients. CONCLUSIONS: Presenting manifestations of meningioma differ between older and younger adult patients, but focality of symptoms and signs is similar. In this case series, the duration of symptoms prior to diagnosis was shorter in older than in younger patients. PMID- 8426033 TI - Nursing home medical directors: ideals and realities. AB - OBJECTIVE: To evaluate what nursing home medical directors actually do, what they and other nursing home personnel believe would be desirable to do, and what problems and deficiencies are perceived. DESIGN: Mail survey with follow-up telephone interview when necessary. SETTING: Forty-five nursing facilities in upstate New York. PARTICIPANTS: The medical directors, administrators, and directors of nursing of the 45 facilities. MEASUREMENTS: Inventory of what medical directors reported as to their actual activities and time spent, and of what they, the administrators, and the directors of nursing felt should be their responsibilities and activities under ideal circumstances. RESULTS: For part-time medical directors, self-reported time spent on medical directorship activities averaged 12 hours per month; of all directors, 45% spent 8 hours or less per month. Proportion of time spent on various specific activities varied widely. There was general agreement that substantially more time should be spent, in particular, on evaluating and addressing problems of adequacy and quality of care, communicating with attending physicians about problems, and assisting with inservice training programs. CONCLUSIONS: To fill the role adequately, more time should be spent by many part-time medical directors, which will require greater financial commitment by facilities and reimbursement systems. Efforts need to made to better coordinate the expectations of medical directors and facility staff. PMID- 8426034 TI - The nursing home medical director role in transition. AB - OBJECTIVE: To compare the self-reported role of nursing home medical directors in 1989 (prior to the implementation of the nursing home reform amendments of OBRA 87 in 1990) with the role outlined in the Joint Commission on Accreditation of Healthcare Organizations Long Term Care Standards Manual. DESIGN: Survey. MEASUREMENTS: Using a telephone interview survey, physicians were questioned about their role as nursing home medical director. Responses were compared with the definition of the medical director's role in the Long Term Care Standards Manual. Information about time spent per month and reimbursement received for medical direction was also obtained. PARTICIPANTS: Physicians serving as nursing home medical directors in 1989 in Houston/Harris County, Texas. RESULTS: Of 50 nursing home medical director positions in Houston/Harris County in 1989, interviews were completed for 36 (72%). The majority of the medical directors met the criteria for having a signed agreement with the facility, assuring continuous medical coverage, developing emergency procedures, and participating in quality assurance activities. Twenty-eight percent of the medical directors received no monetary compensation for their work. Of those who were paid, the mean monthly earnings for nursing home medical direction were $345. CONCLUSION: Although all nursing facilities are now required by regulation to have medical directors, a wide variation in medical directors' level of involvement and commitment was documented. PMID- 8426035 TI - Effects of cognitive impairment on the reliability of geriatric assessments in nursing homes. AB - OBJECTIVE: To explore the relationship between an elderly subject's cognitive status and the reliability of multidimensional assessment data. DESIGN: Survey, with cognitive status as the independent variable and interrater reliability as dependent variable. SETTING: Medicare/Medicaid-certified nursing homes. PARTICIPANTS: 147 residents age 65 or older. MEASUREMENTS: Dual assessments of elderly nursing home residents were performed by nurse assessors using the Health Care Financing Administration's new Minimum Data Set for Nursing Home Resident Assessment and Care Screening (MDS). Assessments were classified on the basis of residents' cognitive status, and levels of disagreement between assessors were analyzed. MAIN RESULTS: Overall assessment reliability, agreement concerning a resident's activities of daily living status, and the reliability of estimates of his or her communication skills and sensory abilities were significantly affected by a resident's cognitive status. The presence of cognitive impairment made these measurements less reliable--especially those related to communication skills, vision, and hearing. CONCLUSIONS: Assessments of residents suffering from cognitive impairment were significantly less reliable than assessments of cognitively intact residents. However, these differences in reliability were not uniform across all assessment domains. When treating the cognitively impaired elderly, clinicians must exercise caution in their reliance on standardized measurements that may be less reliable for this population. PMID- 8426036 TI - Selenium and oxygen-metabolizing enzymes in elderly community residents: a pilot epidemiological study. AB - OBJECTIVE: To investigate the relationship of selenium and oxygen-deactivating enzymes with age in the elderly. SAMPLE: The study sample consisted of volunteers recruited from the PAQUID study. This study is conducted in a representative sample of non-institutionalized individuals aged > or = 65 years living in Southwestern France; its main objective is to study longitudinally the incidence and risk factors of dementia. METHODS: Plasma and erythrocyte selenium and activities of oxygen metabolizing enzymes in erythrocytes (GSH-Px, CuZn-SOD, and GSSG-RD) were measured in 239 volunteers (108 males and 131 females; mean age 73.7 years). RESULTS: Plasma selenium (PSe) decreased significantly with age; a similar but non-significant trend was found for erythrocyte selenium (ESe). None of the enzyme activities showed a clear relationship with age. Women had significantly higher GSH-Px activities than men. For PSe levels lower than 77 ng/mL, there was a strong correlation between PSe and GSH-Px; above this value, the correlation decreased, suggesting that the selenium requirement for GSH-Px production had been satisfied. In this sample, CuZn-SOD was correlated negatively with GSH-Px (r = -0.18; P < or = 0.01) and positively with GSSG-RD (r = +0.20; P < or = 0.01). CONCLUSIONS: In individuals aged > or = 65 years, we found that blood selenium levels were negatively correlated with age. Our analysis of the relationship between selenium and GSH-Px activity suggests that low selenium values are associated with decreased GSH-Px activity. PMID- 8426037 TI - Effects of testosterone replacement therapy in old hypogonadal males: a preliminary study. AB - OBJECTIVE: To examine the effects of testosterone administration to older hypogonadal males (bioavailable testosterone less than 70 ng/dL). DESIGN: Alternate-case controlled trial. SETTING: St. Louis University. PATIENTS: Eight males (mean age 77.6 +/- 2.3 years) who received testosterone and six males (mean age 76 +/- 2.3 years) who served as controls. Selected from alumni of the SHEP trial and attendees at the St. Louis University Impotence Clinic. INTERVENTIONS: Testosterone enanthate (200 mg/mL) was administered intramuscularly to the treatment group every 2 weeks for 3 months. MEASUREMENTS: Serum testosterone, bioavailable testosterone and estradiol, weight, % body fat, right hand muscle strength, balance, cholesterol, HDL-cholesterol, hematocrit, BUN, creatinine, albumin, calcium, PTH, 25(OH) vitamin D, 1,25(OH)2 vitamin D, osteocalcin, prostate-specific antigen, and fructosamine. RESULTS: Males who received testosterone had a significant increase in testosterone and bioavailable testosterone concentration, hematocrit, right hand muscle strength and osteocalcin concentration. They had a decrease in cholesterol (without a change in HDL-cholesterol) levels and decreased BUN/Creatinine ratios. CONCLUSION: These preliminary findings support the need for long term studies of testosterone therapy in older hypogonadal males. PMID- 8426038 TI - Social service interventions for caregivers of patients with dementia: impact on health care utilization and expenditures. AB - OBJECTIVES: An intervention, which had as its primary goal the enhancement of compliance to social work recommendations, was shown to produce extremely high rates of compliance. This report addresses the secondary objective of the study: to evaluate the impact of the intervention on short-term (ie, 6-month) health services utilization and expenditures. DESIGN: Randomized controlled trial. SETTING: University-based memory disorders clinic. PARTICIPANTS: Caregivers of patients with progressive memory disorders. MAIN OUTCOME MEASURES: Service utilization and expenditures. RESULTS: The intervention did not have a statistically significant impact on utilization of either health care or community resources. The intervention group had $903 less expenditures during the study period, a difference that did not achieve statistical significance. The results were consistent when controlling for caregiver characteristics that differed at baseline. CONCLUSIONS: Although the intervention was successful in enhancing compliance with recommendations, more intensive interventions may be required to increase subsequent service utilization. Future investigations may wish to target the appropriateness of services used over a period longer than 6 months. PMID- 8426039 TI - Percutaneous transhepatic cholecystostomy for acute complicated calculous cholecystitis in elderly patients. AB - OBJECTIVE: To assess the immediate and long-term outcomes of elderly patients with acute complicated cholecystitis treated by percutaneous cholecystostomy. To assess the results of bile cultures obtained in this group of patients. DESIGN: Case series. SETTING: Tertiary care center. PATIENTS: Thirty-two patients, with a mean (+/- S.D.) age of 78 +/- 8 years (range, 58-92 years), and who presented with acute cholecystitis complicated by empyema formation. Sixty-six percent had associated disorders, which rendered them at high risk for surgical intervention. INTERVENTION: Percutaneous transhepatic catheter drainage of the gallbladder, with a mean drainage time of 20 days (range 0-84 days). In addition, endoscopic sphincterotomy with removal of common bile duct stones was performed in six patients and percutaneous aspiration of an associated liver abscess in four cases. RESULTS: Percutaneous cholecystostomy was followed by rapid regression of clinical symptoms and of radiologic abnormalities in all patients. Sixteen cases (50%) underwent elective cholecystectomy 1-12 weeks after cholecystostomy. One of them died of aspiration pneumonia, whereas 15 had no post-operative problems and were discharged 9 days (mean) after surgery. Forty-four percent (14/32) were considered inoperable: they remained completely free of biliary symptoms and died of unrelated illness (22%) after a mean follow-up of 6 months (range, 1-22 months) or are still alive (22%) with a mean follow-up of 15 months (range, 5-36 months). Bile cultures were positive in 75% of the patients. Escherichia coli, other aerobic Gram-negative micro-organisms, and anaerobic bacterial species accounted for 35% (16/46), 28% (13/46), and 20% (9/46) of the isolated bacteria, respectively. All aerobic Gram-negative species tested in vitro were susceptible to gentamicin and to temocillin. CONCLUSIONS: Percutaneous transhepatic cholecystostomy is a safe and effective procedure in the treatment of elderly high-risk patients with acute cholecystitis complicated by empyema formation. It can be followed by elective cholecystectomy, if possible, or by expectant conservative management in patients who are inoperable because of systemic disease. PMID- 8426040 TI - Poor outcome of on-site CPR in a multi-level geriatric facility: three and a half years experience at the Baycrest Centre for Geriatric Care. AB - Our objective was to determine the outcome of onsite CPR in elderly patients receiving institutional care who had access to 24-hour on-site full ACLS capability. We used a retrospective chart and CPR protocol review of all patients who received CPR during the study period; these were patients or residents in a multi-level geriatric long-term care facility that provides various degrees of facility-based and ambulatory care. Immediate, short-term, and long-term survivorship were determined and correlated where possible with the category of patient seen (short-term care versus long-term care) and the relationship between the witnessing of the arrest and survival. Of the 41 patients who underwent CPR, there were only four survivors of 60 days or greater. Of these, three subjects were short-term, not long-term care patients of the Centre; they returned to their previous level of function. One fully dependent bed-ridden patient returned to that level of care for a survival period of 100 days. Of the four long-term survivors, three of the arrests were witnessed, and one was indeterminate. No unwitnessed arrests resulted in long-term survival. Our experience suggests that CPR in the elderly long-term care patient is unlikely to be successful even when it is available on-site. Unwitnessed arrests in this population were universally fatal. This information has helped Baycrest Centre for Geriatric Care develop protocols, guidelines, and policies for CPR that are suitable for our population. PMID- 8426041 TI - Lumbar spine disease in the elderly. PMID- 8426043 TI - Research in nursing homes: practical aspects. PMID- 8426042 TI - Interleukin-6: a cytokine for gerontologists. AB - Interleukin-6 (IL-6) is a multifunctional cytokine that presumably plays its major role as a mediator of several of the acute phase inflammatory responses. These include inflammatory cell and lymphocyte activation and hepatocellular stimulation of acute phase protein synthesis. IL-6 expression is normally low, and serum levels are usually non-detectable in the absence of inflammation. However, with advancing age, serum levels become detectable, and it is proposed that this reflects an age-associated loss in the normal regulation of gene expression for this molecule. The cause of this is most likely multi-factorial, but there is evidence that it relates to an age-associated loss of T cell immunoregulatory functions as well as menopausal loss of estrogen. In any event, the "inappropriate" presence of IL-6 results in many changes typical of chronic inflammation. There is also speculation that IL-6 may contribute to the pathogenesis of several diseases of late-life including lymphoma, osteoporosis, and Alzheimer's disease. In this review the biology of this important cytokine is presented and its relevance to gerontology is highlighted. PMID- 8426044 TI - "Lifecare": a viable option for long-term care for the elderly. AB - In the absence of any coherent US long-term care policy, the "lifecare" or "continuing care" retirement community (CCRC) has emerged as one viable model, providing housing, health care, social supports, and long-term care insurance to about 300,000 elderly Americans. Some CCRCs have also demonstrated the cost effectiveness of a health care philosophy targeted to the maximum functional independence (MFI) of residents. Broad dissemination of the CCRC model and the philosophy of MFI depend on development of new approaches to the role of government and private health insurance along with a new understanding of, and commitment to, risk management. PMID- 8426045 TI - Is male hip fracture a marker for low testosterone in elderly male nursing home residents? PMID- 8426046 TI - Reversible dementia caused by vitamin B12 deficiency. PMID- 8426048 TI - Reality orientation boards: a recipe for disorientation? PMID- 8426047 TI - Complications from finger contracture cushions. PMID- 8426049 TI - New OBRA guidelines for nursing facilities. PMID- 8426050 TI - Lewin's manifest dream exercise revisited. AB - In an institute research class, the validity of Lewin's methodology in his manifest dream exercise was tested by a different approach. Class members developed a questionnaire based on two actual manifest dreams. Responses from six analytic candidates and fifteen graduate analysts were analyzed by two classroom groups, working independently. The data were assessed by two methods and the conclusions were only partially synchronous with Lewin's method of collective free association in which members of the group influenced one another. Our subjects did not influence one another, and they and research class members were "blind" to the dreams' associations, context, and meanings derived years earlier, until after data were assessed and conclusions were reached. One conclusion suggests that intuition as to "correct" meanings may be independent of the number of years of analytic experience. This project is reported to stimulate similar research projects as a regular part of psychoanalytic education. PMID- 8426051 TI - "In the neighborhood": aspects of a good interpretation and a "developmental lag" in ego psychology. AB - The significance of the conscious ego in the interpretive process has increasingly come under scrutiny. It is becoming clearer that the analyst's view of the conscious ego orients his interpretive approach, and subtly sets the goals of the analysis. At various times Freud championed the analytic importance of the conscious ego, and alternately rejected its significance. Hartmann's view of the ego stimulated research into a developmental line of the ego, while investigations of the ego in clinical psychoanalysis lagged far behind. The importance of the conscious ego in understanding resistances and levels of communication are explored. PMID- 8426052 TI - The analytic surface. Panel report. PMID- 8426053 TI - The life cycle of the analyst: pregnancy, illness, and disability. Panel report. PMID- 8426054 TI - Stability of gains achieved during analytic treatment from a followup perspective. Panel report. PMID- 8426055 TI - The clinical relevance of the contribution of Winnicott. Panel report. PMID- 8426056 TI - Deterministic chaos and the sciences of complexity: psychoanalysis in the midst of a general scientific revolution. AB - A combination of new concepts and the enormous powers of computation now available have created the beginnings of a major new scientific revolution. It does not have to do with psychoanalysis per se, but with some basic assumptions among the sciences generally. The result is already showing in new visions of nature variously called "deterministic chaos," "nonlinear dynamics," or "sciences of complexity." For the first time it is possible to study complex systems in process, over time. This paper, especially attending to issues of separation and integration, focuses on two components of the new understandings, fractal geometry (part of the mathematics of topology) and deterministic chaos (the tendency of nonlinear systems to oscillate toward and away from absolute chaotic disorganization). In a diverse group of intellectual disciplines, it is now possible to describe systems in operation in detail, in terms of nonlinear differential equations. From these, computer models of multiple variables in interaction can be produced. In turn, this allows experimentation on the models by altering variables. At this time, psychoanalysis can only use deterministic chaos and fractals metaphorically, but in the future, especially if psychoanalysis is seen in terms of process or organismic theory, it is likely that such models can be produced. PMID- 8426057 TI - The current status of psychoanalysis. AB - Presented here is an overview of current challenges and controversies regarding psychoanalysis as a science, competing psychoanalytic theories, convergent and divergent trends in psychoanalytic technique, psychoanalytic education, psychoanalysis as a profession. Among other issues stressed are the importance of the relation of psychoanalysis to the University, the research implications of competing theoretical and technical orientations, the need to reexamine the structure of psychoanalytic education, and the importance of international cross fertilization in expanding the application of psychoanalysis to other fields. PMID- 8426058 TI - Repression: the evolution of a psychoanalytic concept from the 1890's to the 1990's. AB - The author presents a summary of Freud's concept of repression, including modifications of the concept from 1894 to 1932. Several more recent treatments are examined, including those of Piaget; Rosenblatt and Thickstun; Galin; Kissin; Shapiro and Perry; Schwartz; the cognitive and experimental psychologists, including Kihlstrom and Erdelyi; the "connectionists"; and Edelman. Finally, the author delineates several different types of repression, outlining how the different models might apply to the different types of repression. PMID- 8426059 TI - Interpretive fallibility and the psychoanalytic dialogue. AB - The concept of the analyst's interpretive fallibility is utilized to understand and organize the ways in which contemporary psychoanalytic theory has expanded on Freud's notions regarding the hypothetical nature of interpretation. This expansion involves a particular axis regarding the analyst's stance, an axis that cuts across theoretical scaffolding to include the analyst's view of symmetry and asymmetry in the analytic setting and the epistemological positions and preferences accorded to analyst and analysand. The ways in which the analyst constructs and imposes meaning are examined more specifically in the theories of Roy Schafer and Ernest S. Wolf. The benefits and problems of constructivism in analytic stance are evaluated and illustrated through a case example. Finally, the notion of psychoanalytic neurality is reconsidered with regard to a series of tensions between the construction and foreclosure of meaning in the analytic situation. PMID- 8426060 TI - Locomotion and physical development in rats treated with ionizing radiation in utero. AB - Effects of ionizing radiation on the emergence of locomotor skill, and physical development were studied in laboratory rats (Fisher F-344 inbred strain). Rats were treated with 3 different doses of radiation (150 rad, 15 rad, and 6.8 rad) delivered on the 20th day of prenatal life. Results indicated that relatively moderate (15 rad) to high (150 rad) doses of radiation had effects on certain locomotion and physical development parameters. Exposure to 150 rad delayed pivoting, cliff-avoidance, upper jaw tooth eruption, and decreased body weights. Other parameters, such as negative geotaxis, eye opening, and lower jaw tooth eruption were marginally delayed in the 150 rad treated animals. Exposure to 15 rad delayed pivoting and cliff-avoidance. PMID- 8426061 TI - Effect of fungicides, captafol and chlorothalonil, on microbial and enzymatic activities in mineral soil. AB - Tests were conducted to determine the effects of fungicides, captafol and chlorothalonil, on microbial and enzymatic activities in sandy loam. The results indicated that when captafol or chlorothalonil was added to the sandy loam, bacterial and fungicidal populations initially decreased with the treatments but recovered rapidly to levels similar to those in the controls. No inhibition on oxidation of soil ammonia or organic sulfur was observed. The fungicide treatments significantly increased oxygen consumption from the decomposition of organic matter indigenous to the soil. Both fungicides suppressed invertase and amylase for 1 day. However, the inhibitory effect disappeared after 2 days. Captafol depressed dehydrogenase for 4 days and recovered to equal to that of control after 7 days. No inhibitory effect on urease and phosphatase was shown with the fungicidal treatments. Although some stimulatory influences of fungicides on microbial and enzymatic activities were found in the soil, in no instance were the effects dramatic or sufficient enough to be considered important to soil fertility. PMID- 8426062 TI - Acute toxicity of selected organophosphorus pesticides to Cyprinus carpio and Barilius vagra. AB - The acute toxicity of two organophosphorus (OP) pesticides, diazinon and malathion to Cyprinus carpio and Barilius vagra was determined. Mortalities and LC50-96 hr values for B. vagra and C. carpio were variable. Both juvenile common carp and B. vagra were extremely sensitive to diazinon. Adult B. vagra were more sensitive to malathion than were juvenile common carp. PMID- 8426063 TI - Improving preschool-aged vaccination coverage: not business as usual. PMID- 8426064 TI - Commission criticizes fee-for-service system. PMID- 8426065 TI - Health care tops legislative agenda. PMID- 8426066 TI - Thrombolysis in acute myocardial infarction: a hospital emergency room experience. AB - Thrombolysis has become a well-established alternative in the management of acute myocardial infarction. The timing of the initiation of thrombolytic agents is important in determining outcomes. There has been great interest in understanding the delays from onset of symptoms to the initiation of thrombolysis. The experience at Wishard Memorial Hospital in Indianapolis was monitored and reviewed for comparison to other metropolitan area experiences. PMID- 8426067 TI - Health economist stresses coordination of care. Interview by Bob Carlson. PMID- 8426068 TI - Cellular responses of oysters infected with Haplosporidium nelsoni: changes in circulating and tissue-infiltrating hemocytes. AB - The protozoan parasite Haplosporidium nelsoni (MSX) elicits an inflammatory-type response in oysters, Crassostrea virginica. We assayed circulating hemocytes of oysters exposed to H. nelsoni to quantify the effects of parasitism, selection for resistance, and season on total and differential counts. All factors had a significant (P < 0.02) effect on total counts, but explained relatively little of the overall variance [season (12%) > selection (4%) > infection (2%)]. Circulating hemocyte densities increased with infection intensity in resistant animals, but were depressed in susceptible oysters with light infections. Counts in both groups were lowest in August and highest in May, at which times densities in susceptible oysters were reduced (P < 0.02) compared to those in resistant animals. No differences existed in November. Susceptible oysters may be more debilitated by infections than resistant animals, resulting in impaired circulation and depressed cell counts. The number of tissue-infiltrating hemocytes increased with infection intensity (P < 0.0001), but showed no statistically significant (P > 0.05) association with numbers of circulating hemocytes in individual oysters. Size-frequency analysis with a Coulter counter indicated that the proportion of large cells (presumed to be granular hemocytes) was lower (P < 0.0001) in susceptible oysters, which were also heavily infected, compared to resistant oysters, which had very few infections. The loss of granular hemocytes may stem from degranulation associated with tissue damage and inflammation. Present evidence suggests that the principal role of the hemocyte response in MSX disease is to plug lesions, remove debris, and repair tissues and that these functions may help oysters survive infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426069 TI - Lipid and fatty acid analysis of the Plodia interpunctella granulosis virus (PiGV) envelope. AB - Virus envelope was isolated from Plodia interpunctella granulosis virus, produced in early fourth-instar larvae. Both polar and neutral lipids were analyzed by two dimensional thin-layer chromatography. Fatty acid composition of various individual neutral and polar lipids was determined by gas-liquid chromatography. The major components of envelope neutral lipid were diacylglycerols. Palmitic acid and stearic acid were the major saturated fatty acids in both polar and neutral lipids. Whereas palmitoleic acid was the major unsaturated fatty acids in neutral lipids, oleic acid was the major unsaturated fatty acid in the polar lipids. PMID- 8426070 TI - Natural history of atherosclerosis and hyperlipidemia in heterozygous WHHL (WHHL Hh) rabbits. II. Morphologic evaluation of spontaneously occurring aortic and coronary lesions. AB - This study evaluated the morphologic appearance of spontaneous aortic and coronary atherosclerotic lesions in 21 of the 28, 3-year old, heterozygous Watanabe heritable hyperlipidemic (WHHL-Hh) rabbits whose lipid profiles were presented in part I of this report. In situ perfusion fixation of the arteries showed 100% of the aortas involved with one or another type of intimal lesion. In male rabbits (n = 13), the abdominal aortas had more severe fibrosis and more diffuse intima thickening than the thoracic aortas, (p < 0.001). In female rabbits (n = 8), fatty streaks and fibrous lesions were more predominant in the thoracic than in the abdominal aorta (p < 0.05). Fatty aortic plaques in the female were more commonly found in the abdominal than in the thoracic aorta, but this finding was not statistically significant. In contrast, fatty aortic plaques were not found in the male aortas; however, larger areas of diffuse intimal thickening with fatty deposits were more common in the abdominal aortas of the males (p < 0.01). Annular arteriosclerotic lesions, exclusive to the thoracic aorta, occurred in three of 21 rabbits. Histologically, the aortas demonstrated subintimal fibrosis, fragmentation of the lamina elastica interna, focal medial degeneration, and cholesterol clefts. The coronary arteries were involved in 85% of the males and in 86% of the females. Lesions were more common in the left than in the right coronary artery and primarily consisted of mild nonobstructing intimal hyperplasia with fibrosis. Based on these observations, we consider older WHHL-Hh rabbits excellent models for studying atherosclerosis. Because of their genetic and age-related lesions, WHHL-Hh rabbits may be superior to the cholesterol-fed rabbit model with respect to comparability with human atherosclerotic lesions. PMID- 8426071 TI - G-protein and drug trafficking on islet crossroads: implications for stress and diabetes. PMID- 8426072 TI - Lysophosphatidylcholine accumulation in ischemic human myocardium. AB - Lysophosphatidylcholine accumulates in the coronary sinus during pacing-induced myocardial ischemia in humans. This amphiphile accelerates Ca++ flux leading to cell injury in cultured cardiac myocytes, but it is not known whether lysophosphatidylcholine accumulation is injurious to human myocardium. In this study, we measured lysophosphatidylcholine in normal human myocardium obtained during cardiac surgery and exposed to ischemic conditions in vitro. Total lysophosphatidylcholine concentration (sum of lysophosphatidylcholine remaining in tissue and lysophosphatidylcholine released into the buffer) increased from 0.73 +/- 0.08 nmol/mg protein at baseline to 1.83 +/- 0.45 nmol/mg protein after 5 minutes of ischemia (p < 0.001), and was associated with evidence of cell injury (26% depletion of tissue lactate dehydrogenase). Significant lysophosphatidylcholine release into the incubation buffer (0.41 +/- 0.11 nmol/mg protein) also occurred after 5 minutes of ischemia. In contrast, there was no lysophosphatidylcholine accumulation or release and no lactate dehydrogenase depletion in oxygenated and perfused controls. Attenuation of lysophosphatidylcholine accumulation by incubation with lysophospholipase did not prevent cell injury. Lysoplasmalogen was not detected in ischemic tissue. We conclude that lysophosphatidylcholine accumulation is a marker of myocardial ischemia in humans. PMID- 8426073 TI - Upregulation of Mac-1 surface expression on neutrophils during simulated extracorporeal circulation. AB - The leukocyte integrin Mac-1 (alpha m beta 2, CD11b/18 CR3, MO1), in addition to binding iC3b, has been shown to be the receptor for the coagulation proteins fibrinogen, factor X, and high molecular weight kininogen. Mac-1 is known to be upregulated by agonists that stimulate neutrophils or monocytes. Previous studies from this laboratory have documented neutrophil activation during cardiopulmonary bypass. We therefore used an experimental model for cardiopulmonary bypass, a simulated extracorporeal circulation, to study the surface expression of Mac-1 on peripheral blood leukocytes by immunofluorescence flow cytometry. The number of Mac-1 receptors in polymorphonuclear leukocytes had increased 2.2 times and 2.9 times baseline by 2 hours at 37 degrees C and 28 degrees C, respectively (p < 0.001). Neutrophil elastase-alpha 1-proteinase inhibitor complexes (a measure of neutrophil degranulation) increased more than sixfold from 41 micrograms/L to 256 micrograms/L after 2 hours at 28 degrees C. The number of Mac-1 receptors expressed on polymorphonuclear leukocytes correlated with polymorphonuclear leukocyte elastase complexes (r = 0.93). Mac-1--bearing lymphocytes did not display any change in receptor number in the simulated extracorporeal circulation. By 2 hours, monocytes at 37 degrees C showed only insignificant increase; at 28 degrees C they displayed a 1.6 times increase (p < 0.05). By maintaining a temperature-matched standing control samples, we could prove that, unlike the Mac-1 in polymorphonuclear leukocytes, this increase in monocyte Mac-1 is not from bypass but is rather an effect of temperature. These data suggest that a receptor for procoagulant proteins on circulating polymorphonuclear leukocytes (Mac-1) is upregulated during simulated extracorporeal circulation. PMID- 8426074 TI - Iron loading of cultured cardiac myocytes modifies sarcolemmal structure and increases lysosomal fragility. AB - The mechanism of damage to myocardial subcellular organelles was studied in iron loaded rat myocardial cells in culture in an attempt to identify the primary target of iron's toxic effects. Lysosomes and sarcolemmal membranes were purified by fractionation of the postnuclear supernatant on a 6.7% colloidal polyvinylpyrrolidone-coated silica gradient. After 24-hour incubation with ferric ammonium citrate at a concentration of 20 micrograms/ml (0.36 mmol/L) iron, a selective depletion of polyunsaturated fatty acids was found in whole-cell homogenates, as well as in the postnuclear supernatant and sediment. Iron loading resulted in a sharp increase in the total activity of the lysosomal enzyme beta hexosaminidase in unfractionated whole-cell homogenates, increased free enzyme activity, and loss of latent activity indicating increased lysosomal fragility. Conversely, iron loading resulted in a marked decrease in the activity of the sarcolemmal enzyme 5'-nucleotidase and a significant loss of total protein sulfhydryl group content. These studies in cultured heart cells are in agreement with previous observations indicating increased lysosomal fragility in iron loaded hepatic and splenic tissues, attributed to increased membrane lipid peroxidation. In addition, the marked decrease in sarcolemmal 5'-nucleotidase activity and in total protein sulfhydryl group content imply that iron-induced peroxidative damage to membrane proteins may be a more important mechanism in the pathogenesis of altered myocardial function in the iron-loaded heart than formerly was recognized. PMID- 8426075 TI - Plasminogen activation in hereditary angioneurotic edema. PMID- 8426076 TI - Stabilization of platelet-fibrinogen interactions: modulation by divalent cations. AB - The binding of fibrinogen to its GPIIb-IIIa receptor is divalent-cation dependent. In addition to Ca+2 and Mg+2, Mn+2 has been shown to modulate adhesive protein interactions with integrins. This study examined the effect of Mn+2 on fibrinogen interactions with intact platelets. Compared with that of control platelets in buffer containing 1 mmol/L Mg+2, fibrinogen binding to adenosine diphosphate- or thrombin-stimulated platelets decreased 23% +/- 12% and 15% +/- 9% (mean +/- SD, n = 4), respectively, after addition of 1 mmol/L Mn+2. No change in binding affinity was noted, but the stability of platelet-fibrinogen interactions was diminished markedly. Ethylenediaminetetraacetic acid dissociated 68% +/- 8% of fibrinogen bound to ADP-treated platelets (p < 0.05) during a 60 minute incubation with fibrinogen and 1 mmol/L Mn+2, compared with 40% +/- 13% of fibrinogen bound to control platelets and 29% +/- 8% of fibrinogen bound in the presence of Ca+2 (mean +/- SD, n = 6). Mn+2 also diminished the stabilization of fibrinogen interaction with thrombin-stimulated platelets and inhibited the recovery of bound fibrinogen with the Triton X-100 (Union Carbide Corp., Danbury, Conn.) insoluble cytoskeleton. Only 31% +/- 10% of fibrinogen bound to thrombin stimulated platelets for 60 minutes in the presence of Mn+2 associated with the cytoskeleton (p < 0.05), compared with 61% +/- 14% and 75% +/- 20% of fibrinogen bound to control platelets incubated with and without Ca+2, respectively. Mn+2 further inhibited large adenosine diphosphate- or thrombin-induced platelet aggregate formation and reduced the ability of platelets to retract fibrin clots. These data suggest that Mn+2 alters GPIIb-IIIa function relative to native fibrinogen and support a role for the stabilization of platelet-fibrinogen interactions in platelet aggregation and clot retraction. PMID- 8426077 TI - Hyperglucagonemia and hyperdynamic circulation in rats with biliary cirrhosis. AB - Glucagon, a hormone with a potent vasodilatory action, has been suggested to play a role in the hyperkinetic circulation in animals with portocaval shunts and chronic portal venous stenosis; however, its role in cirrhosis has not been examined. We tested the hypothesis that increased levels of circulating glucagon contribute to the hyperdynamic circulatory state found in rats with biliary cirrhosis. Compared with plasma glucagon levels in sham-operated controls, glucagon levels, measured by radioimmunoassay, were significantly increased in chronic bile duct-ligated (5 to 6 weeks) cirrhotic rats (603 +/- 107 pg/ml vs 197 +/- 11 pg/ml, p < 0.001) but not in acute bile duct-ligated (10 days) cholestatic rats (310 +/- 55 pg/ml). Unanesthetized, catheter-implanted cirrhotic--but not cholestatic--rats exhibited a hyperdynamic circulatory state with increased cardiac index and decreased systemic vascular resistance. The plasma glucagon level correlated with cardiac index (r = 0.62, p < 0.01) and systemic vascular resistance (r = -0.54, p < 0.05). In addition, cirrhotic rats showed depressed hypoxic pulmonary vasoconstriction that was correlated with increased plasma glucagon level (r = -0.68, p < 0.01). We conclude that circulating glucagon is increased in rats with biliary cirrhosis and that hyperglucagonemia may contribute to the observed systemic hyperdynamic circulatory state and the loss of hypoxic pulmonary vasoconstriction in these rats. PMID- 8426078 TI - Signal transduction, second messengers, and platelet function. PMID- 8426079 TI - The hormonal and metabolic responses to prolonged hypoglycemia. AB - The purpose of this review was to summarize some of the recent advances that have been made in the understanding of the physiology involved in the counter regulatory response to prolonged hypoglycemia. It is hoped that this review may stimulate thought and increase awareness that other factors, in addition to the glycemic level, can affect the counterregulatory response. However, if the goal of tight metabolic control is to be achieved in subjects with diabetes, the spectre of severe hypoglycemia must be removed. Thus further work is needed to understand the physiologic mechanisms controlling hypoglycemic counterregulation in normal subjects and the syndromes of abnormal counterregulatory responses present in subjects with IDDM. PMID- 8426080 TI - Generation of plasmin during acute attacks of hereditary angioedema. AB - Hereditary angioedema is caused by a genetic deficiency of C1-inhibitor, a serine protease inhibitor that regulates activation of complement, contact, and fibrinolytic systems. Symptoms (bouts of subcutaneous and mucous swelling) depend on the release of a vasoactive mediator, probably through activation of these three systems. We studied the interrelationship among complement, contact, and fibrinolytic activation in 23 patients with hereditary angiodema, 18 during remission and five during an attack, by measuring plasma levels of C1-C1 inhibitor, factor XIIa-C1 inhibitor, kallikrein-C1 inhibitor, and plasmin-alpha 2 antiplasmin complexes, tissue plasminogen activator, and urokinase plasminogen activator. In addition, cleavage of high-molecular weight kininogen was detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis and quantified by densitometry. During remission, plasma levels of C1-C1 inhibitor complexes were elevated (p = 0.0002), whereas the other parameters were within the normal range. During acute attacks, not only plasma levels of C1-C1 inhibitor complexes but also those of plasmin-alpha 2-antiplasmin complexes (P = 0.0009) and cleaved high-molecular weight kininogen were elevated. A positive correlation between plasmin-alpha 2-antiplasmin complexes and cleaved high-molecular weight kininogen was observed (r = 0.75, p < 0.001). This article presents the first in vivo evidence that supports the concept that release of vasoactive mediators in hereditary angiodema attacks is associated with the activation of the fibrinolytic system. PMID- 8426081 TI - Concomitant defect in internal release and influx of calcium in patients with congenital platelet dysfunction and impaired agonist-induced calcium mobilization. Thromboxane production is not required for internal release of calcium. AB - The rise in cytoplasmic ionized calcium concentration ([Ca2+]i) on platelet activation is a combination of Ca2+ release from internal stores and influx of extracellular Ca2+. To understand the underlying mechanisms, we studied internal release and influx of Ca2+ in platelets from four patients with impaired agonist induced Ca2+ mobilization and abnormal platelet aggregation and secretion responses. In normal platelets, thrombin caused a dose-dependent increase in internal release and influx; aspirin inhibited the total rise in [Ca2+]i and influx but not internal release, indicating that internal release occurs independent of cyclooxygenase products. In the four patients, both internal release and influx of Ca2+ induced by thrombin and adenosine diphosphate were diminished; the defect was more striking at lower agonist concentrations. To determine whether the Ca2+ storage organelles of these platelets had a diminished responsiveness to inositol 1,4,5-triphosphate (IP3) we studied IP3 (0.05 to 5 mumol/L) induced Ca2+ release and found it to be normal in all patients. We conclude that the impaired Ca2+ mobilization in our patients is due to abnormalities in both internal release and influx and that it is unlikely to be due to impaired platelet responsiveness to IP3 or defective thromboxane production. The impaired Ca2+ mobilization may be due to defects in phospholipase C activation and IP3 production. These patients provide direct evidence that internal release and influx of Ca2+ on platelet activation are closely interrelated. PMID- 8426082 TI - Modulation of intrinsic prothrombin activation by fibrinogen and fibrin I. AB - Recent observations raise the possibility that by binding thrombin, fibrin(ogen) could modulate prothrombin activation, perhaps by modulating thrombin-mediated activation of factors VIII and V. Factors VIIIa and Va are the cofactors of intrinsic tenase and prothrombinase, respectively. This study compared the profile of prothrombin activation in pooled normal plasma, defibrinated pooled normal plasma, and pooled normal plasma containing fibrin I. Prothrombin activation was initiated with three stimuli, as follows: (1) addition of a suspension of thrombin, CaCl2, and coagulant phospholipids to plasma; (2) contact activation followed by recalcification of plasma; and (3) addition of a suspension of crude rabbit brain tissue factor and CaCl2 to plasma. Each plasma sample contained 2 mmol/L gly-pro-arg-pro, which prevents the polymerization of fibrin. Prothrombin activation, indexed as the concentration of prothrombin activation fragment 1 + 2 produced, was quantitated by enzyme-linked immunosorbent assay. Fibrinogen significantly delayed the onset of prothrombin activation initiated by thrombin or by contact activation but not that initiated with tissue factor. In contrast, fibrin I in solution accelerated prothrombin activation initiated with thrombin or by contact activation. Although fibrinogen had no effect on intrinsic activation of factor IX or factor IX activation initiated by adding thrombin to plasma, it inhibited factor X activation initiated with either stimuli. We therefore conclude that fibrinogen can inhibit the activation of factor X and prothrombin. Second, fibrin I accelerates prothrombin activation. Fibrinogen and fibrin, therefore, have the potential to bioregulate blood coagulation. PMID- 8426083 TI - Mesangial proliferative glomerulonephritis induced in rats by a lentil lectin and its antibodies. AB - Experimental glomerulonephritis was induced in rats to investigate the consequence of the antigen-antibody interaction on the surface of glomerular endothelial cells (GENs). A lectin, Lens culinaris hemoagglutinin (LCH), was first planted in the left kidney by isolated perfusion of a left kidney, and then the circulation was reestablished. Rabbit anti-LCH antibodies were injected from the tail vein 3 minutes after the recirculation of the left kidney, and acute glomerulonephritis ensued. Fifteen minutes after the injection, rabbit immunoglobulin G (IgG), rat C3, and LCH were observed exclusively on the surface of GENs. Accumulation of platelets was prominent. Three hours later, the immune deposits were seen in the subendothelial space, and the polymorphonuclear cells were the dominant infiltrate in the glomeruli. Up to the seventh day, immune deposits were seen in the subendothelial space, and the widening of this area was increasingly observed. Fourteen days later, immune deposits containing rat IgG were observed in the subepithelial area, but they were only occasionally seen in the subendothelial space and in the mesangial area. No crescent formation was seen at day 14, but the mesangial area was expanded, with an increased number of cells. The number of nuclei in the cross-section of a glomerulus increased after the induction of glomerulonephritis, but the number of leukocyte common antigen positive cells (infiltrating cells) decreased gradually from day 4 to day 14. The staining of Thy-1.1, a marker of mesangial cell, was markedly enlarged in the glomerulus at day 14. These data suggest that mesangial proliferative glomerulonephritis can be induced by the antigen-antibody interaction on the surface of GENs. PMID- 8426084 TI - Fibrinopeptide A in liver cirrhosis: evidence against a major contribution of disseminated intravascular coagulation to coagulopathy of chronic liver disease. AB - To test the hypothesis that disseminated intravascular coagulation contributes to hemostatic failure in liver cirrhosis, fibrinopeptide A and fibrin(ogen) degradation fragment E were measured in 69 patients with stable liver cirrhosis and compared with fibrinopeptide A and fibrin(ogen) degradation fragment E in 32 healthy subjects, 33 patients with thromboembolism, and 10 patients with hypofibrinogenemic disseminated intravascular coagulation. Mean fibrinopeptide A in cirrhosis was slightly increased compared with healthy subjects (2.4 vs. 1.8 ng/ml, p < 0.005), but fourfold lower than in thromboembolism (mean fibrinopeptide A 9.7 ng/ml; p < 0.0001), and tenfold lower than in disseminated intravascular coagulation (mean FPA 24.3 ng/ml; p < 0.0001). Single fibrinopeptide A levels in cirrhosis were within the normal range in 75% of the patients, marginally increased in 9%, and definitely increased in 16%. A definite increase in both fibrinopeptide A and fibrin(ogen) degradation fragment E, which characterized the groups of patients with thromboembolism and disseminated intravascular coagulation, was found in 10% of the cirrhotic patients. Among 17 patients with cirrhosis and hypofibrinogenemia, mean fibrinopeptide A (2.7 ng/ml) was tenfold lower compared with mean fibrinopeptide A in patients with hypofibrinogenemic disseminated intravascular coagulation (p < 0.0001), whereas the frequency of increased single fibrinopeptide A levels (29%) was not significantly different compared with the 52 cirrhotic patients without hypofibrinogenemia (single levels elevated in 23% of the cases). Moreover, the frequency of hypofibrinogenemia, thrombocytopenia, or abnormal clotting times was not significantly different in cirrhotic patients with normal fibrinopeptide A level when compared with cirrhotic patients with increased fibrinopeptide A. These findings do not support an important contribution of disseminated intravascular coagulation to coagulopathy of liver cirrhosis. PMID- 8426085 TI - Electrocardiographic patterns in V4R lead in experimental right ventricular infarction of rats. AB - Twenty-four male rats that became hypertensive after complete ligature of the abdominal aorta, just above the origin of the left renal artery, showed combined myocardial infarction. To study the electrocardiographic patterns of the experimental right ventricular infarction, the V4R right thoracic lead was recorded at different time intervals after aortic ligature. The patterns recorded were as follows: Q waves in 23 cases (95.8%), ST-T segment elevation in 11 cases (45.8%), ST-T segment depression in two cases (8.3%), and decrease in voltage of R wave in four cases (16.6%), with a predominance of the alterations of the ST-T segment in the acute phase and the appearance of Q waves during the subacute phase. PMID- 8426086 TI - Natural history of atherosclerosis and hyperlipidemia in heterozygous WHHL (WHHL Hh) rabbits. I. The effects of aging and gender on plasma lipids and lipoproteins. AB - Plasma lipid and lipoprotein levels are influenced by physiologic changes during the life span of mammalian organisms. In the present study, the effects of aging and gender on plasma lipids and lipoprotein cholesterol fractions in heterozygous Watanabe heritable hyperlipidemic (WHHL-Hh) rabbits were examined. For 3 years the rabbits were fed regular rabbit chow and were free from any experimental manipulation. Male WHHL-Hh rabbits (n = 17) had a 55% increase in total plasma cholesterol level (p < 0.005) between the ages of 7.5 and 34 months. This increase was due to significant increases in very low-density lipoprotein cholesterol (VLDL-C) (28%, p < 0.05), in low-density lipoprotein cholesterol (126%, p < 0.005), and in high-density lipoprotein cholesterol (28%, p < 0.05). Female WHHL-Hh rabbits (n = 11) did not show significant differences in lipid plasma values with aging. However, their plasma values, with the exception of the VLDL-C, were significantly (p < 0.05) higher than those of the male rabbits. Pregnancy and increased weight in the older female rabbits may have accounted for these differences. Significant plasma lipid changes occur as a function of heterozygosity, age, and gender in WHHL-Hh rabbits. The lipoprotein changes seen in male but not in female WHHL-Hh rabbits during the aging process are similar to those observed in human subjects. Pregnancy in WHHL-Hh rabbits, like in human beings, may induce hyperlipidemia, which is opposite to the effects observed in normal rabbits or homozygous WHHL rabbits. PMID- 8426087 TI - Immunotherapy of tularemia: characterization of a monoclonal antibody reactive with Francisella tularensis. AB - An IgM monoclonal antibody (mAb) recognized surface antigens specific to Francisella tularensis wild-type (Schu4) and live vaccine strain (LVS), and reacted with both in ELISA and slide agglutination tests. This mAb also reacted with LVS microorganisms in tissues of infected mice as assessed by an indirect fluorescence technique. Western blot analysis showed the mAb to react with antigens associated with F. tularensis LPS. PMID- 8426088 TI - Differentiation of dendritic cell populations in macrophage colony-stimulating factor-deficient mice homozygous for the osteopetrosis (op) mutation. AB - In op/op mice, immunohistochemical and electron microscopic techniques were used to examine the effects of the OP mutation on dendritic cell populations in lymphoid tissues and skin. In the thymic medulla, T cell zone of lymph nodes, and splenic white pulp of op/op mice, numbers of NLDC-145-positive dendritic cells were not decreased. Compared to the normal littermates, numbers of BM8-positive macrophages were reduced in various tissues of the mutant mice, including the lymphoid tissues. These dendritic cells of op/op mice expressed Ia antigens but not F4/80 and BM8 antigens. Ultrastructurally, the dendritic cells developed a tubulovesicular system typical of interdigitating cells, but they were abnormal in that interdigitation of their cytoplasmic processes was not prominent. In the epidermis of the op/op mice, dendritic cells expressed NLDC-145, F4/80, Ia antigens, and adenosine diphosphatase or adenosine triphosphatase activity, and numbers of NLDC-145-, Ia-, or ADPase-positive dendritic cells were reduced slightly, but these reductions were not significant statistically. Birbeck granules were detected in most of them electron microscopically. These results indicate that nonlymphoid dendritic cells develop in the lymphoid tissues and skin of op/op mouse, suggesting that they are differentiated from granulocyte macrophage colony-forming cells or earlier hematopoietic cell precursors. PMID- 8426089 TI - Induction of a 26-kDa membrane-form tumor necrosis factor (TNF)-alpha in human alveolar macrophages. AB - The expressions of a membrane form of TNF (m-TNF) by human alveolar macrophages (AM) and autologous blood monocytes from healthy donors were examined. Upon lipopolysaccharide (LPS) stimulation, AM produced 26-kDa TNF-alpha on their cell surface. We designed a bioassay for measuring m-TNF in which macrophages were fixed with paraformaldehyde after stimulation for 18 h, then m-TNF activity was assessed as cytotoxicity of fixed macrophages on L929 cells. This assay was specific to m-TNF because: 1) no soluble factors were contributed to the cytotoxicity of fixed AM, 2) anti-TNF-alpha monoclonal antibody completely neutralized m-TNF activity, and 3) m-TNF activity was not altered after low-pH or high-salt treatment. On LPS stimulation, AM produced significant amounts of m-TNF earlier than TNF-alpha secretion. AM also expressed significant amounts of m-TNF when stimulated with other bacterial components and their derivatives. Interleukin (IL)-4 suppressed both m-TNF production and TNF-alpha secretion. p Toluene-sulfonyl-L-arginine methyl ester (TAME) inhibited specifically TNF-alpha secretion and not m-TNF expression. Although blood monocytes produced small amounts of m-TNF, monocyte-derived macrophages showed enhanced m-TNF after cultivation with GM-CSF for 10 days. These findings indicate that m-TNF is expressed as a step in the TNF-alpha producing system, and suggest that m-TNF may play important roles in exhibition of macrophage function in situ. PMID- 8426090 TI - Three distinct cell phenotypes of induced-TNF cytotoxicity and their relationship to apoptosis. AB - We have identified three distinct cell phenotypes with respect to the conditions under which cells became susceptible to TNF-mediated lysis. These conditions include: 1) treatment with the protein synthesis inhibitor, cycloheximide; 2) contact with activated macrophages, and 3) infection with vaccinia virus. Whereas vaccinia virus-infected 3T3 cells became sensitive to soluble TNF, F5b cells required contact with activated macrophages. We showed that the "macrophage resistant" F5m cells did not become sensitive to TNF or to killing by activated macrophages after infection with vaccinia virus. Therefore, vaccinia infection does not sensitize all cells to TNF. We also determined the pathways of lysis for cells after sensitization. Whereas 3T3, LM929, and F5b cells were killed by the process of necrosis, F5m cells lysis was characterized by the release of low mol wt DNA fragments (apoptosis). PMID- 8426091 TI - Selective refractoriness of macrophages to endotoxin-induced production of tumor necrosis factor, elicited by an autocrine mechanism. AB - Tolerance of macrophages to endotoxin (lipopolysaccharide, LPS) can be induced in vitro by LPS itself. We show here that one of the mechanisms of tolerance to LPS can be mediated via an autocrine process. Continuous exposure to LPS is not required to induce macrophage desensitization. Refractoriness to production of tumor necrosis factor (TNF) in response to LPS can be transferred from tolerant to naive macrophage populations by incubation of the latter with the culture supernatant of the former, in the absence of endotoxin. The active factor present in this macrophage-desensitizing culture supernatant (MD-Sup) is more efficiently removed by incubation with tolerant macrophages than by incubation with naive macrophages. The refractoriness elicited by treatment with MD-Sup is restricted to a decreased TNF response to LPS; interleukin-1 (IL-1) and IL-6 responses are not affected. PMID- 8426092 TI - Activation of tumoricidal properties in macrophages by lipopolysaccharide requires protein-tyrosine kinase activity. AB - The purpose of these studies was to determine whether triggering murine peritoneal macrophages to a tumoricidal state by lipopolysaccharide (LPS) requires protein-tyrosine phosphorylation. The LPS-triggered activation of mouse macrophages to lyse syngeneic B16 melanoma cells was significantly inhibited in a dose-dependent manner by the protein-tyrosine kinase (PTK) inhibitors genistein, herbimycin A, and tyrphostin. Genistein was effective only when added to macrophages prior to or simultaneously with LPS. Genistein potently inhibited the productive interaction of macrophages with LPS but had only a minor effect on the action of interferon-gamma. The effects of genistein on LPS-triggered macrophage activation were not due to nonspecific changes in macrophage metabolism or toxicity because genistein did not prevent lysis of tumor cells by activated macrophages, nor did it reduce the capacity of macrophages to phagocytose antibody-opsonized sheep erythrocytes. Western blot analysis with antiphosphotyrosine monoclonal antibody revealed that incubation of macrophages with LPS produced a rapid increase in tyrosine phosphorylation of several proteins and that the induced phosphorylation could be inhibited by effective concentrations of genistein, herbimycin A, or tyrphostin. Taken together, these data indicate that protein-tyrosine phosphorylation plays an important role in LPS-induced tumoricidal activation of macrophages. PMID- 8426093 TI - Effects of modulators of cytosolic Ca2+ on phytohemagglutin-dependent Ca2+ response and interleukin-2 production in Jurkat cells. AB - We have previously reported the presence, in Jurkat T cells, of outward K+ currents and inward currents that have been attributed to Ca2+ channels. Here, we have studied the effects of dimethyl 1,4-dihydro-2,6-dimethyl-4-(2-nitrophenyl) 3,5-pyridine-dicarboxylate (nifedipine) and 4-(2,1,3-benzoxadiazol-4-yl)-1,4 dihydro-2,6-dimethyl-5- methoxy-carbonylpyridine-3-carboxylate (PN200-110), two dihydropyridines (DHPs) known to inhibit voltage-dependent Ca2+ channel activity in different types of cells, and two inhibitors of internal Ca2+ release (muscle cells), ryanodine and 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester (TMB-8), on the Phaseolus vulgaris phytohemagglutinin (PHA)-dependent responses in Jurkat T lymphocytes. Our results show that nifedipine and PN200-110 inhibit the PHA-dependent production of interleukin-2 except when 12-O-tetradecanoyl-13-O acetyl phorbol is added to the cultures. Ryanodine and TMB-8 are not inhibitors. The PHA-dependent Ca2+ response is significantly reduced when the cells are preincubated in the presence of the DHPs. Under these conditions, ryanodine has only a small inhibitory effect and TMB-8 has no effect. In contrast, only ryanodine (50 microM) decreases the PHA-dependent cytosolic release of Ca2+i when the cells are bathed in a medium containing a low concentration of Ca2+ (60 nM). The inhibitory effects of nifedipine and PN200-110 may result from the binding of these DHPs to specific receptor sites as revealed by studies using [3H]PN200-110 (KD = 8.5 +/- 3.1 nM; 2300 +/- 500 apparent binding sites/cell). Photoaffinity labeling studies using [3H]azidopine as a probe showed specific incorporation of label into three glycoproteins of molecular mass (+/- SD) 170 +/- 13, 110 +/- 25, and 60 +/- 17 kd as analyzed by electrophoresis under reducing conditions. PMID- 8426094 TI - Activation of primary lymphocytes requires prolonged lectin stimulation. AB - T lymphocytes are activated by a complex series of events, but the mechanisms remain unclear. One uncertainty is the time of receptor-ligand interaction necessary for commitment to DNA synthesis and proliferation. Although this issue has broad implications for the interpretation of T cell activation data, it remains unresolved. Therefore, we examined the temporal activation requirements of rat splenocytes stimulated with concanavalin A (Con A) by measuring proliferation, as well as interleukin-2 (IL-2) production and IL-2 receptor IL 2R) expression. Splenocytes stimulated with various Con A concentrations for 3 h did not incorporate significantly more [3H]thymidine than unstimulated splenocytes. Some increase occurred after 6 h of lectin exposure but maximum proliferation occurred only after the 52-h stimulation. Furthermore, Con A incubations of 6 h or more were required for significant increases in IL-2 or IL 2R. Maximum lymphokine production and receptor expression were observed after the 52-h stimulation. Thus, activation of some primary lymphocytes required only 6 h of stimulation, but much longer mitogen contact was necessary for maximum recruitment. PMID- 8426096 TI - The medical Gulag. PMID- 8426095 TI - Fibronectin cell-binding domain triggered transmembrane signal transduction in human monocytes. AB - Fibronectin (Fn) fragments have recently been shown to stimulate tumor necrosis factor (TNF) secretion by human monocytes. In this study, we investigated the signal transduction mechanisms involved in Fn-induced TNF secretion. Treatment of human monocytes with Fn120, a chymotryptic cell-binding fragment of plasma Fn, failed to cause a detectable rise in Ca2+ mobilization. Fn120-induced TNF secretion could be inhibited with Ca2+ channel blockers. The protein kinase C (PKC) inhibitors H-7 and sphingosine inhibited the TNF-inducing activity of Fn120. HA1004 was used as a control for the isoquinoline sulfonamide derivatives and did not change Fn120-induced TNF secretion by monocytes. H-8 inhibited TNF secretion at higher concentrations. A calmodulin-dependent kinase inhibitor, W-7, was found to be effective, with 50% inhibition of Fn120-induced TNF secretion at 5 microM. The activation and translocation of PKC were measured directly. In unstimulated monocytes, approximately 70% of PKC activity was found in the cytosol and 30% in the membrane. Following the stimulation of monocytes with phorbol myristate acetate (100 nM), rapid and sustained translocation of PKC from the cytosol to the membrane was observed. The stimulation of monocytes with Fn120 triggered a rapid translocation of PKC within 2 to 5 min, followed by a return to normal levels within 8 min. These findings support the conclusion that Fn120 induced TNF secretion requires the activation of PKC. PMID- 8426097 TI - How to make a successful transition into private practice. PMID- 8426098 TI - Altering records: discrediting your best witness. AB - There is a proper way to make a correction to a medical record. If the correction is entered properly, contemporaneously, has bearing on current patient care, does not embellish or amplify your actions as being correct, and is consistent with other entries in the record it should be sound. Entries that are not properly made can destroy a professional liability defense. Plaintiff's attorneys will look for a reason to allege altered records. It is a plaintiff's attorney's strategy to use such an allegation to make the case less defensible, force a settlement or increase the value of a case through the threat of punitive damages. Do not give them the opportunity. PMID- 8426099 TI - Conducting occupational medical examinations under the new ADA. PMID- 8426100 TI - The CARE Program: what it is. PMID- 8426101 TI - Clinton S.C., between two wars: a story. PMID- 8426102 TI - The diagnosis of wide complex tachycardia. AB - Ventricular tachycardia is the diagnosis in approximately 80% of cases of WCT and in approximately 95% of cases with structural heart disease. The presence of atrioventricular dissociation, fusion beats, capture beats, and concordance are important findings strongly suggesting VT; whereas, QRS frontal axis and ventricular rate provide less definitive diagnostic information. A QRS width greater than 160 msec. for LBBB configuration WCT or greater than 140 msec. for RBBB configuration argues strongly for VT. The QRS morphology may be very helpful in selected cases. The duration of the arrhythmia does not discriminate between the etiologies of WCT, as all rhythms may be hemodynamically stable for prolonged periods. Finally, response to treatment may give diagnostic information in addition to therapeutic benefit. PMID- 8426103 TI - Cardiac rehabilitation--against all odds. PMID- 8426104 TI - Thymic origin of embryonic intestinal gamma/delta T cells. AB - Current evidence suggests both thymic and extrathymic origins for T cells. Studies in mice favor an in situ origin for a prominent population of intestinal intraepithelial lymphocytes that express gamma/delta T cell receptor (TCR). This developmental issue is explored in an avian model in which the gamma/delta lymphocytes constitute a major T cell subpopulation that is accessible for study during the earliest stages of lymphocyte development. In the chick embryo, cells bearing the gamma/delta TCR appear first in the thymus where they reach peak levels on days 14-15 of embryogenesis, just 2 d before gamma/delta T cells appear in the intestine. Using two congenic chick strains, one of which expresses the ov antigen, we studied the origin and kinetics of intestinal colonization by gamma/delta T cells. The embryonic gamma/delta+ thymocytes homed to the intestine where they survived for months, whereas an embryonic gamma/delta- thymocyte population enriched in thymocyte precursors failed to give rise to intestinal gamma/delta+ T cells. Embryonic hemopoietic tissues, bone marrow, and spleen, were also ineffective sources for intestinal gamma/delta+ T cells. Intestinal colonization by gamma/delta+ thymocytes occurred in two discrete waves in embryos and newly hatched birds. The data indicate that intestinal gamma/delta T cells in the chicken are primarily thymic migrants that are relatively long-lived. PMID- 8426105 TI - Identification of human cancers deficient in antigen processing. AB - Intracellular antigens must be processed before presentation to CD8+ T cells by major histocompatibility complex (MHC) class I molecules. Using a recombinant vaccinia virus (Vac) to transiently express the Kd molecule, we studied the antigen processing efficiency of 26 different human tumor lines. Three cell lines, all human small cell lung carcinoma, consistently failed to process endogenously synthesized proteins for presentation to Kd-restricted, Vac-specific T cells. Pulse-chase experiments showed that MHC class I molecules were not transported by these cell lines from the endoplasmic reticulum to the cell surface. This finding suggested that peptides were not available for binding to nascent MHC molecules in the endoplasmic reticulum. Northern blot analysis of these cells revealed low to nondetectable levels of mRNAs for MHC-encoded proteasome components LMP-7 and LMP-2, as well as the putative peptide transporters TAP-1 and TAP-2. Treatment of cells with interferon gamma enhanced expression of these mRNAs and reversed the observed functional and biochemical deficits. Our findings suggest that downregulation of antigen processing may be one of the strategies used by tumors to escape immune surveillance. Potential therapeutic applications of these findings include enhancing antigen processing at the level of the transcription of MHC-encoded proteasome and transporter genes. PMID- 8426106 TI - Pfs2400 can mediate antibody-dependent malaria transmission inhibition and may be the Plasmodium falciparum 11.1 gene product. AB - Monoclonal antibodies (mAb) have been raised against Plasmodium falciparum gametocyte stage protein extracts, in an effort to identify novel parasite antigens that might mediate malaria transmission-blocking immunity. mAb 1A1 identified Pfs2400, a sexual stage-specific antigen of greater than 2 megadaltons, that is associated with the outer leaflet of the parasitophorous vacuole membrane in mature circulating gametocyte-infected red blood cells. Upon induction of gametogenesis, Pfs2400 partitions between the gamete plasmalemma and the degenerating erythrocyte membrane. The antigen is no longer detectable in the fully emerged gamete. mAb 1A1 dramatically reduces the number of oocysts formed in P. falciparum gametocyte-fed mosquitoes. The cognate antigen is probably the product of the Pf11.1 gene (Scherf et al. 1988. EMBO [Eur. Mol. Biol. Organ.]J. 7:1129) on the basis that a peptide composed of two copies of the degenerate nine amino acid repeat sequence in the Pf11.1 protein, can inhibit binding of mAb1A1 to the native antigen. The mechanism of transmission inhibition mediated by the Pfs2400 is discussed. PMID- 8426107 TI - Restricted immunoglobulin junctional diversity in neonatal B cells results from developmental selection rather than homology-based V(D)J joining. AB - The mechanism by which coding ends are joined during immunoglobulin (Ig) recombination is poorly understood. Recently, short sequence similarities (2-6 bp) observed at the ends of certain variable (V), diversity (D), and joining (J) gene segments of Ig have been correlated with limited junctional diversity observed in coding exons assembled from these elements. However, it is unclear whether these sequence homologies play any direct role in favoring coding joint formation by influencing the V(D)J recombination process. In this report, we demonstrate that coding sequence similarities do not influence the position of coding joints during V(D)J recombination in vivo. Instead, during embryonic development, B cells with certain joining products undergo progressive selection. Developmental selection is completed before exposure to external antigens and appears to be determined by the amino acid sequence encoded by the coding joint. We conclude that the nucleotide sequences of the coding regions do not play a major role in directing V(D)J recombination. Instead, we propose that limited Ig junctional diversity results from prenatal developmental selection of B cells based on the protein sequence of their surface Ig antigen-binding site. Sequence identities at the ends of coding segments may have evolved because they increase the likelihood that a selectable antigen-binding site is created during a random recombination process. PMID- 8426108 TI - Tolerance of CD8+ T cells developing in parent-->F1 chimeras prepared with supralethal irradiation: step-wise induction of tolerance in the intrathymic and extrathymic environments. AB - Tolerance of CD8+ cells was examined in parent-->F1 bone marrow chimeras (BMC) prepared with supralethal irradiation; host class I expression in the chimeras was limited to non-BM-derived cells. In terms of helper-independent proliferative responses in vitro and induction of graft-vs.-host disease on adoptive transfer, CD8+ cells from long-term chimeras showed profound tolerance to host antigens irrespective of whether the cells were prepared from the thymus or from spleen or lymph nodes. By limiting dilution analysis, cytotoxic T lymphocyte (CTL) precursors specific for host antigens were rare in the extrathymic lymphoid tissues. In the thymus, by contrast, host-specific CTL precursors were only slightly less frequent than in normal parental strain mice. These host-specific CD8+ cells survived when BMC thymocytes were transferred intravenously to a neutral environment, i.e., to donor strain mice. When transferred to further BMC hosts, however, most of the host-reactive cells disappeared. Collectively, the data suggest that tolerance of CD8+ cells in BMC hosts occurs in both the intrathymic and extrathymic environments. In the thymus, contact with host antigens on thymic epithelial cells deletes CD8+ cells controlling helper independent proliferative responses and in vivo effector functions but spares typical helper-dependent CTL precursors. After export from the thymus, most of the CTL precursors are eliminated after contacting host antigens on stromal cells in the extrathymic environment. PMID- 8426109 TI - Exclusion of circulating T cells from the thymus does not apply in the neonatal period. AB - Although T cells arise in the thymus, migration of mature postthymic T cells back to the thymus is very limited in adult mice and is restricted to activated cells. In neonates, by contrast, we present evidence that circulating CD4+ and CD8+ T cells with a naive/resting phenotype readily enter the thymus after intravenous injection and remain there for prolonged periods. The migration of resting T cells to the neonatal thymus is largely limited to an unusual subset of cells which lacks expression of the lymph node homing receptor, leukocyte-endothelial cell adhesion molecule 1 (LECAM-1) (MEL-14). Migration of mature T cells to the thymus in neonates may be important for self-tolerance induction. PMID- 8426110 TI - Downregulation of the antigen presenting cell function(s) of pulmonary dendritic cells in vivo by resident alveolar macrophages. AB - Class II major histocompatibility complex (Ia)-bearing dendritic cells (DC) from airway epithelium and lung parenchyma express low-moderate antigen presenting cell (APC) activity when freshly isolated. However, this function is markedly upregulated during overnight culture in a manner analogous to epidermal Langerhans cells. The in vitro "maturation" process is inhibited by coculture with pulmonary alveolar macrophages (PAM) across a semipermeable membrane, and the degree of inhibition achieved can be markedly increased by the presence of tumor necrosis factor alpha. In addition, PAM-mediated suppression of DC function is abrogated via inhibition of the nitric oxide synthetase pathway. Functional maturation of the DC is accompanied by increased expression of surface Ia, which is also inhibited in the presence of PAM. Prior elimination of PAM from DC donors via intratracheal administration of the cytotoxic drug dichloromethylene diphosphonate in liposomes, 24-72 h before lung DC preparation, achieves a comparable upregulation of APC activity, suggesting that (consistent with the in vitro data) the resident PAM population actively suppresses the APC function of lung DC in situ. In support of the feasibility of such a regulatory mechanism, electron microscopic examination of normal lung fixed by intravascular perfusion in the inflated state (which optimally preserves PAM in situ), revealed that the majority are preferentially localized in recesses at the alveolar septal junctions. In this position, the PAM are in intimate association with the alveolar epithelial surface, and are effectively separated by as little as 0.2 microns from underlying interstitial spaces which contain the peripheral lung DC population. A similar juxtaposition of airway intraepithelial DC is demonstrated with underlying submucosal tissue macrophages, where the separation between the two cell populations is effectively the width of the basal lamina. PMID- 8426111 TI - High-frequency representation of a single VH gene in the expressed human B cell repertoire. AB - Idiotype (Id) 16/6 marks a variable (V) region structure that occurs frequently in the human immunoglobulin repertoire. The basis of the Id has been traced to a germline heavy chain gene segment, VH18/2 (VH26). To pursue the molecular basis for the frequency of Id 16/6, we have analyzed polymerase chain reaction generated C mu, C gamma, and VH3 family V gene libraries derived from the circulating and tonsillar B cells of four normal individuals and from the B cells of two patients with active systemic lupus erythematosus (SLE). The frequency of VH18/2 in these libraries was compared with three control VH genes, VH56P1, VH21/28, and VHA57. Plaque lifts from C mu and C gamma VH cDNA libraries were screened with gene-specific oligonucleotide probes. The frequency of VH18/2 ranged from 4 to 10% of JH+ plaques (two of five times that of control VH genes). In four VH3 family-specific libraries derived from rearranged DNA, VH18/2 represented 19-33% of VH3+ plaques. Hybridizing VH18/2 plaques were 98-100% homologous to the germline VH gene; mutations when present were often in framework 3. Extensive variation was seen in the complementarity determining region 3 sequences of these rearranged V genes. The high frequency of VH18/2 expression in the B cell repertoire was confirmed by sequencing randomly picked JH+ plaques. In two patients with active SLE the frequency of use of VH18/2 was not greater than that observed in normal subjects. These results show that VH18/2 is overrepresented in the B cell repertoire of normal subjects and suggest that the immune repertoire may be dominated by relatively few V genes. PMID- 8426112 TI - Linkage of pemphigus vulgaris antibody to the major histocompatibility complex in healthy relatives of patients. AB - Pemphigus vulgaris (PV) is an autoimmune disease caused by high concentrations of antibody to an epidermal cadherin. The disease is associated with two kinds of HLA-DR4, DQ8 haplotypes dominantly distributed among Jewish patients, and these plus DR6, DQ5 haplotypes in non-Jewish patients. Low levels of the PV antibody were found in 48% of a total of 120 asymptomatic parents, children, and siblings of 31 patients, thus exhibiting dominant inheritance. The inheritance of these low levels of antibody in asymptomatic relatives was linked to the major histocompatibility complex with a highly significant logarithm of the odds score of 9.07, almost always to a DR4 or DR6 haplotype of the patient. Disease appears to occur in susceptible individuals with low levels of antibody when a second factor, either environmental or genetic, induces high levels, sufficient to produce blisters. PMID- 8426113 TI - Prevention of tumor metastasis formation by anti-variant CD44. AB - A splice variant of CD44 (CD44v) originally discovered on metastases of a rat pancreatic adenocarcinoma (BSp73ASML) has been shown by transfection to confer metastatic behavior to nonmetastatic tumor cells (Gunthert U., M. Hofmann, W. Rudy, S. Reber, M. Zoller, I. Haussmann, S. Matzku, A. Wenzel, H. Ponta, and P. Herrlich. 1991. Cell. 65:13). A monoclonal antibody (mAb), 1.1ASML, to the metastasis-specific domain of the CD44v molecule retards growth of lymph node and lung metastases of the metastatic tumor line BSp73ASML, and can efficiently prevent formation of metastases by the transfected line. The antibody is only effective when given before lymph node colonization. Anti-CD44v does not downregulate the expression of CD44v, and prevention of metastatic growth by anti CD44v is not due to activation of any kind of immune defense. We suggest that the mAb interferes with proliferation of metastasizing tumor cells in the draining lymph node, most probably by blocking a ligand interaction. The interference with metastatic spread will greatly facilitate the exploration of the function of CD44v and, in particular, may also open new strategies for the therapy of human metastases. PMID- 8426114 TI - In vivo role of interleukin 4 in T cell tolerance induced by aqueous protein antigen. AB - High doses of aqueous protein antigens induce a form of immunological tolerance in which interleukin 2 (IL-2)- and interferon gamma (IFN-gamma)-secreting T helper type 1 (Th1) cells are inhibited, but IL-4-secreting (Th2) cells are not. This is manifested by reduced proliferation of antigen-specific T cells upon in vitro restimulation, and marked suppression of specific antibody responses of the immunoglobulin (Ig)G2a, IgG2b, and IgG3 isotypes, but not of IgG1 and IgE. The role of the immunomodulatory cytokine IL-4 in this model of unresponsiveness to protein antigens has been examined. Administration of tolerizing antigen itself primes splenic CD4+ T cells for secretion of lymphokines, both IL-2 and IL-4. Neutralization of IL-4 in vivo with the anti-IL-4 antibody 11B11 during tolerance induction augments IFN-gamma production by T cells of tolerant mice, and reverses the suppression of IgG2a, IgG2b, and IgG3. This blockade of IL-4 function does not, however, restore the proliferative responses of T cells, suggesting that reduced T cell proliferation is due to direct T cell inactivation or anergy. Inhibiting the activity of IL-4 in vivo also inhibits the expansion of antigen specific Th2-like cells, which are resistant to the induction of unresponsiveness. Thus, the immunologic consequences of high-dose tolerance are due to a combination of clonal T cell anergy and IL-4-mediated immune regulation. PMID- 8426115 TI - Characterization of a novel trans-sialidase of Trypanosoma brucei procyclic trypomastigotes and identification of procyclin as the main sialic acid acceptor. AB - Here we report the presence of a trans-sialidase on the surface of Trypanosoma brucei culture-derived procyclic trypomastigotes. The enzyme is not detected in lysates of bloodstream trypomastigotes enriched for either stumpy or slender forms. The trans-sialidase catalyzes the transfer of alpha(2-3)-linked sialic acid residues to lactose. beta-galactopyranosyl residues are at least 100 times better acceptors for sialic acid than alpha-galactopyranosyl residues. In the absence of efficient acceptors, the purified enzyme transfers sialic acid to water, i.e., it acts as a sialidase. Although the T. cruzi and T. brucei trans sialidases have very similar donor and acceptor specificities, they are antigenically distinct. Sodium dodecyl sulfate-polyacramide gel electrophoresis under nonreducing conditions and silver staining of the purified trans-sialidase reveals a single band of 63 kD. When the surface membrane of live procyclic trypomastigotes is trans-sialylated, using radioactive sialyllactose as the donor substrate, it appears that the only sialylated surface molecule is procyclin. Pronase treatment of live parasites removes only part of the surface sialic acid, in agreement with recent data showing that the glycosylphosphatidylinositol anchor of procyclin is sialylated (Ferguson, M. A. J., M. Murray, H. Rutherford, and M. J. McConville. 1993. Biochem. J. In press). PMID- 8426116 TI - Activation of cytokine genes in T cells during primary and secondary murine influenza pneumonia. AB - The patterns of cytokine mRNA expression in mice with primary or secondary influenza pneumonia have been assessed by in situ hybridization analysis of cells from both the mediastinal lymph node (MLN) and the virus-infected lung. Evidence of substantial transcriptional activity was found in all lymphocyte subsets recovered from both anatomical sites. The kinetics of cytokine mRNA expression after primary infection with an H3N2 virus were in accord with the idea that the initial response occurs in regional lymphoid tissue, with the effector T cells later moving to the lung. This temporal separation was much less apparent for the more rapid secondary response resulting from challenge of H3N2-primed mice with an H1N1 virus. Among the T cell receptor alpha/beta+ subsets, transcripts for interferon (IFN) gamma and tumor necrosis factor beta were most commonly found in the CD8+ population whereas mRNA for interleukin (IL) 4 and IL-10 was much more prevalent in CD4+ T cells. The gamma/delta T cells expressed mRNA for all cytokines tested, with IL-2, IL-4, and IFN-gamma predominating among those recovered from the inflammatory exudate. At particular time points, especially early in the MLN and late in the infected lung, the frequency of mRNA+ lymphocytes was much higher than would be expected from current understanding of the prevalence of virus-specific precursors and effectors. If this response is typical, induction of cytokine gene expression for T cells that are not responding directly to the invading pathogen may be a prominent feature of acute virus infections. PMID- 8426117 TI - Somatic hypermutation of an immunoglobulin mu heavy chain transgene. AB - We have analyzed somatic hypermutation of an immunoglobulin (Ig) heavy chain transgene. Hybridomas expressing the transgene were produced from immunized transgenic mice and transgene copies were sequenced to assay for mutation. In two IgM-producing hybridomas, as well as in several IgG-producing hybridomas, mutations were found in the VDJ region of the transgene. In the IgM-producing hybridomas, both mutated and unmutated transgene copies were present and expressed as mRNA. Several mutated transgene copies were present in a single cell and these showed different patterns of mutation. Two IgG-producing hybridomas isolated from a single animal also showed a hierarchical pattern of mutation indicating that transgene mutations can accumulate during B cell proliferation, similar to the mutational process for endogenous antibody genes. Among hybridomas that expressed both IgG and IgM molecules derived from the transgene, the isotype switched gamma transgene copy exhibited a higher level of mutation than the mu transgene copies. Our results indicate that the 15-kb ARSmu transgene contains all the sequence information required to target the Ig-specific hypermutational machinery, and raise the possibility that sequences associated with the endogenous CH locus might enhance somatic mutation. PMID- 8426118 TI - Pgs28 belongs to a family of epidermal growth factor-like antigens that are targets of malaria transmission-blocking antibodies. AB - Although Pgs28, a 28-kD surface protein of Plasmodium gallinaceum ookinetes, was previously thought not to be a target of transmission-blocking antibodies, we found that polyclonal antisera to Pgs28 completely blocked parasite infectivity to Aedes aegypti mosquitoes. Antisera raised against reduced Pgs28 were less effective in blocking transmission than were antisera to nonreduced Pgs28; thus, the target epitope(s) of transmission-blocking antibodies appears to be conformation dependent. In stage-specific assays, polyclonal antisera impaired the in vitro transformation of zygotes to mature ookinetes, as well as the in vivo development of mature ookinetes to oocysts. Using microsequence of immunoaffinity-purified Pgs28, we cloned the 666-bp open reading frame of the Pgs28 gene. The deduced amino acid sequence of Pgs28 is strikingly similar to that of a P. gallinaceum zygote surface protein, Pgs25, and its P. falciparum analogue, Pfs25. Pgs28, like Pgs25 and Pfs25, has a presumptive secretory signal sequence, followed by four epidermal growth factor-like domains, and a terminal hydrophobic region. PMID- 8426120 TI - Deficient biosynthesis of N-acetylglucosaminyl-phosphatidylinositol, the first intermediate of glycosyl phosphatidylinositol anchor biosynthesis, in cell lines established from patients with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is a hemolytic disorder caused by a deficiency of biosynthesis of the glycosyl phosphatidylinositol (GPI) anchor, but the biochemical defect is not completely understood. In the present study, we have analyzed affected cell lines established recently from two Japanese patients with PNH. Two lines of evidence indicate that these cells do not synthesize N acetylglucosaminyl-phosphatidylinositol, the first intermediate in the GPI anchor biosynthesis. First, somatic cell hybridization analysis using Thy-1-deficient murine thymoma cell lines with known biochemical defects as fusion partners showed that the PNH cell lines belong to complementation class A, which is known not to synthesize N-acetylglucosaminyl-phosphatidylinositol. Second, analysis of in vitro glycolipid biosynthesis demonstrated that cell lysates of these PNH cell lines in fact did not support biosynthesis of N-acetylglucosaminyl phosphatidylinositol. Thus, we have characterized for the first time the exact biochemical defect leading to PNH. PMID- 8426119 TI - Lipopolysaccharide-induced selective priming effects on tumor necrosis factor alpha and nitric oxide production in mouse peritoneal macrophages. AB - Preculture of thioglycollate-elicited C3HeB/FeJ mouse peritoneal macrophages in vitro with subthreshold stimulatory concentrations of lipopolysaccharide (LPS) can induce hyporesponsiveness (desensitization) to both tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO) production when these cells are subsequently stimulated with 100 ng/ml of LPS. We have established, however, that the primary dose of LPS required for inducing downregulation of NO production is significantly lower than that required for inducing downregulation of TNF-alpha production. Further, when LPS-pretreated macrophages become refractory to subsequent LPS stimulation for NO production, the secondary LPS-stimulated TNF alpha production is markedly enhanced, and vice versa. These results indicate that LPS-induced TNF-alpha and NO production by macrophages are differentially regulated, and that the observed desensitization process may not reflect a state in which macrophages are totally refractory to subsequent LPS stimulation. Rather, our data suggest that LPS-pretreated macrophages become selectively primed for differential responses to LPS. The LPS-induced selective priming effects are not restricted to LPS stimulation, but extend as well to stimuli such as zymosan, Staphylococcus aureus, and heat-killed Listeria monocytogenes. PMID- 8426121 TI - Interferon gamma inhibits interleukin 10 production by monocytes. AB - Interleukin 10 (IL-10) was first described for its ability to inhibit interferon gamma (IFN-gamma) production. Herein, we studied the balance between IFN-gamma and IL-10 production by human peripheral blood mononuclear cells (PBMC) in response to Staphylococcus aureus Cowan (SAC) or lipopolysaccharide (LPS). Monocyte depletion reduced IL-10 production by 90% and resulted in an increased IFN-gamma production. Addition of anti-IL-10 antibody to PBMC cultures also strongly increased IFN-gamma production. In contrast, among various cytokines, only IFN-gamma strongly reduced IL-10 synthesis by SAC- or LPS-activated PBMC and monocytes. Thus, IFN-gamma has proinflammatory effects through the combination of two mechanisms: (a) induction of early tumor necrosis factor alpha (TNF-alpha) and IL-1 beta synthesis; and (b) inhibition of the delayed production of IL-10, an inhibitor of TNF-alpha and IL-1 beta synthesis. Taken together, the present data indicate that IFN-gamma and IL-10 antagonize each other's production and function. PMID- 8426122 TI - Glutamic acid decarboxylase 67-reactive T cells: a marker of insulin-dependent diabetes. AB - Glutamic acid decarboxylase (GAD) has been shown to be a target of autoantibodies in insulin-dependent diabetes (IDD). Two forms of GAD, with molecular weights of 67,000 and 65,000, have been cloned from separate genes. As pancreatic islet beta cell destruction DD is an autoimmune process mediated by T cells, we sought to determine if recombinant GAD67 was recognized by T cells in IDD subjects and particularly their first-degree relatives with islet cell antibodies known to be at risk for IDD. The central regions of human islet and brain GAD67 (amino acids 208-404) were cloned as fusion proteins with glutathione-S-transferase (GST). Proliferation of peripheral blood T cells in the presence of recombinant GAD67 was significantly higher in both at-risk relatives and recent-onset IDD subjects than in other autoimmune disease subjects and human histocompatibility leukocyte antigen (HLA)-matched healthy controls. Thus, 12 of 29 (41%) at-risk relatives and 11 of 29 (38%) recent-onset IDD subjects responded to GAD67, compared with 1 of 7 (14%) other autoimmune disease subjects and 1 of 23 (4%) HLA-matched controls. T cell responses to GST alone or to tetanus toxoid were not different between the groups. These findings demonstrate that GAD67 is a target autoantigen of T cells in IDD and suggest the possibility that GAD-reactive T cells may delineate asymptomatic subjects at increased risk for IDD. PMID- 8426123 TI - T cell receptor ligation induces interleukin (IL) 2R beta chain expression in rat CD4,8 double positive thymocytes, initiating an IL-2-dependent differentiation pathway of CD8 alpha+/beta- T cells. AB - The role of interleukin (IL)2 in intrathymic T cell development is highly controversial, and nothing is known about IL-2R expression on thymocytes of the T cell receptor (TCR) alpha/beta lineage undergoing TCR-driven differentiation events. We analyze here IL-2R alpha and beta mRNA expression in an in vitro system where newly generated rat CD4,8 double positive (DP) thymocytes respond to TCR ligation plus IL-2 (but not to either stimulus alone) with rapid differentiation to functional CD8 single positive T cells (Hunig, T., and R. Mitnacht. 1991. J. Exp. Med. 173:561). TCR ligation induced expression of IL-2R beta (but not alpha) chain mRNA in DP thymocytes. Addition of IL-2 then lead to functional maturation and expression of the IL-2R alpha chain. To investigate if the CD8 T cells generated via this IL-2R beta-driven pathway in vitro correspond to the bulk of CD8 T cells seeding peripheral lymphoid organs in vivo, we compared their phenotype to that of lymph node CD8 T cells. Surprisingly, analysis of CD8 cell surface expression using a novel anti-CD8 monoclonal antibody specific for the alpha/beta heterodimeric isoform, and of CD8 alpha and beta chain mRNA revealed that T cells generated by TCR ligation plus IL-2 resemble thymus-independent rather than thymus-derived CD8 cells in that they express CD8 alpha without beta chains. These findings demonstrate that TCR crosslinking induces functional IL-2R on immature DP rat thymocytes. In addition, they show that at least in vitro, CD8 alpha/alpha T cells are generated from TCR stimulated DP thymocytes (which express the CD8 alpha/beta in the heterodimeric isoform) along an IL-2-driven pathway of T cell differentiation. PMID- 8426124 TI - Interleukin 10 reduces the release of tumor necrosis factor and prevents lethality in experimental endotoxemia. AB - Because of its ability to efficiently inhibit in vitro cytokine production by activated macrophages, we hypothesized that interleukin (IL) 10 might be of particular interest in preventing endotoxin-induced toxicity. We therefore examined the effects of IL-10 administration before lipopolysaccharide (LPS) challenge in mice. A marked reduction in the amounts of LPS-induced tumor necrosis factor (TNF) release in the circulation was observed after IL-10 pretreatment at doses at low as 10 U. IL-10 also efficiently prevented the hypothermia generated by the injection of 100 micrograms LPS. Finally, pretreatment with a single injection of 1,000 U IL-10 completely prevented the mortality consecutive to the challenge with 500 micrograms LPS, a dose that was lethal in 50% of the control mice. We conclude that IL-10 inhibits in vivo TNF secretion and protects against the lethality of endotoxin in a murine model of septic shock. PMID- 8426125 TI - In vivo induction of interleukin 10 by anti-CD3 monoclonal antibody or bacterial lipopolysaccharide: differential modulation by cyclosporin A. AB - We investigated the in vivo effects of cyclosporin A (CsA) on the production of interleukin (IL) 10, a cytokine with major immunosuppressive properties. To elicit IL-10 production in vivo, BALB/c mice were injected either with the anti mouse CD3 145-2C11 monoclonal antibody (mAb) (25 micrograms) or with bacterial lipopolysaccharide (LPS) (20 micrograms). A systemic release of IL-10 was observed in both models, IL-10 serum levels reaching 1.60 +/- 0.32 U/ml (mean +/- SEM) and 0.67 +/- 0.09 U/ml 6 h after injection of 145-2C11 mAb and LPS, respectively. Experiments in nude mice indicated that T cells are involved in the induction of IL-10 by anti-CD3 mAb, but not by LPS. Pretreatment with CsA (total dose: 50 mg/kg) before injection of 145-2C11 mAb completely prevented the release of IL-10 in serum as well as IL-10 mRNA accumulation in spleen cells. In contrast, CsA markedly enhanced LPS-induced IL-10 release (IL-10 serum levels at 6 h: 8.31 +/- 0.43 vs. 0.71 +/- 0.15 U/ml in mice pretreated with CsA vehicle control, p < 0.001), as well as IL-10 mRNA accumulation in spleen. We conclude that CsA differentially affects IL-10 production in vivo depending on the nature of the eliciting agent. This observation might be relevant to clinical settings, especially in organ transplantation. PMID- 8426126 TI - An allelic polymorphism within the human tumor necrosis factor alpha promoter region is strongly associated with HLA A1, B8, and DR3 alleles. AB - The tumor necrosis factor (TNF) alpha gene lies within the class III region of the major histocompatibility complex (MHC), telomeric to the class II and centromeric to the class I region. We have recently described the first polymorphism within the human TNF-alpha locus. This is biallelic and lies within the promoter region. Frequency analysis of the TNF-alpha polymorphism, using the polymerase chain reaction and single-stranded conformational polymorphism, in HLA typed individuals, reveals a very strong association between the uncommon TNF allele and HLA A1, B8, and DR3 alleles. This is the first association between TNF alpha and other MHC alleles and raises the possibility that the uncommon TNF alpha allele may contribute to the many autoimmune associations of the A1,B8,DR3 haplotype. PMID- 8426127 TI - Family planning. PMID- 8426129 TI - Physician anger. PMID- 8426128 TI - Physician anger. PMID- 8426130 TI - Coding and reimbursement. PMID- 8426131 TI - D&HH persons. PMID- 8426132 TI - Universal neonatal hepatitis B immunization--are we jumping on the bandwagon too early? PMID- 8426133 TI - Family physician acceptance of universal hepatitis B immunization of infants. AB - BACKGROUND: The incidence of hepatitis B infection has risen 37% over the last decade; 300,000 new infections and 5000 deaths occur annually in the United States. Because immunization programs that targeted high-risk groups failed to abate this increase, the Centers for Disease Control (CDC) recommended in November 1991 universal hepatitis B immunization of infants. Details were published in an addendum to Morbidity and Mortality Weekly Report. The purpose of this study was to assess (1) the effectiveness of the CDC in disseminating a new immunization recommendation to family physicians, (2) the effect of the new recommendation on clinical practice, and (3) the degree to which noneconomic barriers may affect adoption of universal hepatitis B immunization. METHODS: A random sample of 300 family physicians in North Carolina was surveyed by mail. Descriptive statistics and chi-square analysis were used to assess the relationship of variables hypothesized to predict physician awareness of, and agreement with, the new recommendation. RESULTS: The response rate was 78%. Overall, 48% of family physicians who administered immunizations to children were aware of the new hepatitis B vaccine recommendation. However, only 17% agreed that it was warranted for all newborns in their practice. Twenty-five percent expected more than one half of the parents to refuse three injections at a single well-child visit, a result of adding this vaccine to the current primary immunization schedule. Additionally, 42% expected nurses to resist giving three injections at one visit. CONCLUSIONS: The CDC does not have an effective mechanism for disseminating information to all physicians who care for children. Improved coordination of recommendations between the CDC and relevant specialty societies may help to increase physician adoption of new immunization recommendations in their clinical practice. Additionally, practical concerns of physicians and their patients regarding multiple injections and other practice relevant issues must be considered when formulating new immunization recommendations, if their implementation is to be successful. Additional research is needed to determine effective methods to disseminate immunization information and to address practical concerns of clinicians. PMID- 8426134 TI - Validity of immunization documentation presented to a student health program. AB - BACKGROUND: Unavoidable exposure to disease and to patients susceptible and vulnerable to disease warrants that students entering medical school be immunized against many of the illnesses for which vaccines are available. The validity of immunization records presented at the time of registration, however, is largely dependent on the provision of accurate and reliable documentation by the student. METHODS: We evaluated for authenticity the immunization and tuberculin testing records of 85 students entering medical school in 1990. Five levels of valid documentation were defined, and the information on each record was reviewed accordingly. RESULTS: Only 43% of the records were original documents or laboratory reports of antibody titers, and 7.5% were not date-specific. We found that 8% to 20% of the forms were missing physician and/or student signatures, and 12% to 19% of the forms did not have health care provider addresses. CONCLUSIONS: Even though medical student preventive health programs may have strict requirements, there may be substantial deficiencies in the quality of the documentation provided by the students. Such deficiencies undermine the purpose of these programs. PMID- 8426135 TI - Physicians' attitudes and behaviors regarding hepatitis B immunization. AB - BACKGROUND: Hepatitis B virus (HBV) infections are a serious health problem in the United States, where approximately 18,000 cases are reported each year to the Centers for Disease Control. Even among health care providers, reported vaccination rates have ranged from 17% to 68%. The purpose of this study was to determine the important factors influencing physicians in an academic medical center to receive the HBV vaccination since the introduction of recombinant yeast derived vaccines. METHODS: The 1282 house staff and attending physicians in a university medical center were surveyed regarding their HBV vaccination history. The characteristics of vaccinated and nonvaccinated physicians and their attitudes and concerns regarding vaccination were compared. RESULTS: Of the 813 physicians who responded, 54.0% had been vaccinated. Vaccination rates varied with level of training, from 91.9% among first- and second-year residents to 32.2% among attending physicians. Although physicians in specialties at higher risk for infection were more likely to have been vaccinated, only 40.0% of pathologists and 51.9% of obstetrician-gynecologists reported having been vaccinated. Using multivariate analysis, we found important demographic predictors of HBV vaccination included physician sex and years since graduation, as well as level of training and specialty. Physicians who had been offered the vaccine were more likely to have been vaccinated. CONCLUSIONS: These results show that many physicians in an academic medical center, particularly those at an early stage of their training, have received HBV vaccination. Our results suggest that programs offering hepatitis B vaccinations to physicians can be effective in reducing this group's risk of hepatitis B infections. Special efforts may be necessary to reach physicians who have completed their training. PMID- 8426136 TI - Women's satisfaction with birth control. AB - BACKGROUND: Contraception is a major component of preventive health care for women. There are indications that women are not satisfied with the methods of birth control currently available. Dissatisfaction with contraceptive methods may lead to unplanned pregnancies. METHODS: Adult women visiting the family health center over a 1-year period were invited to participate in a research interview. Questions were asked about demographic variables and the women's use of, and their satisfaction with, contraceptive methods. RESULTS: Many women were displeased with the present methods of birth control. This is reflected in the numerous methods used by each woman, and by frequent use of permanent sterilization as a contraceptive method. Women were as dissatisfied with oral contraceptives as they were with the less efficacious methods such as condoms, foams, gels, and rhythm. The only methods that had a greater than 70% satisfaction rate were tubal ligations and partner's vasectomies. CONCLUSIONS: There is significant dissatisfaction with the methods of contraception currently available. Increased patient-physician discussion and education may improve satisfaction with birth control methods now used. New methods of contraception may be needed to prevent unplanned pregnancies. PMID- 8426137 TI - Usefulness of antimicrosomal antibody titers in the diagnosis and treatment of postpartum thyroiditis. AB - BACKGROUND: Postpartum thyroiditis is a common but frequently unrecognized disorder, affecting approximately 5% of women during the first 12 months after delivery. We investigated whether the antimicrosomal antibody titer could be used to determine which women with positive titers postpartum (1) might develop symptomatic or biochemical abnormalities within the first postpartum year (early disease), (2) might require therapy with thyroid hormone, and (3) might have persistent abnormalities (late disease). METHODS: Women (n = 55) who had positive antimicrosomal antibody titers at delivery were prospectively followed for 11 to 45 months. Titers were evaluated again at 6 to 10 weeks postpartum and approximately every 8 weeks for the first year. RESULTS: Early disease occurred in 40 of 55 (73%) women, late disease occurred in 29 of 55 (53%) women, and treatment was required by 21 of 55 (38%) women. The occurrence of early disease was associated with the occurrence of late disease (P < .05). The chances of developing early disease were 6 to 1 (P = .01) when serum titers of antimicrosomal antibodies were > or = 400 at delivery, and 5 to 1 (P = .02) when titers were > or = 1600 at 6 to 10 weeks postpartum. The chances of being given thyroid hormone therapy were 23 to 1 (P = .006) when titers at delivery were > or = 6400, and 6 to 1 when titers at 6 to 10 weeks postpartum were > or = 6400 (P = .004). Titers were not useful in estimating who would have late disease. CONCLUSIONS: Screening for postpartum thyroid dysfunction after delivery using antimicrosomal antibody titers is highly useful. The titer value can help guide the physician in the care of patients with postpartum thyroiditis whose disease may not be self-limiting and who will probably require thyroid hormone therapy. PMID- 8426138 TI - Identifying and treating wife abuse. AB - BACKGROUND: Wife abuse, acknowledged as a critical problem in our society, is often undetected by family physicians. The purpose of this study was to identify the problems and potential solutions encountered by family physicians in the identification and treatment of wife abuse in London, Ontario. METHODS: Family physicians in London were recruited to participate in four focus groups. The groups' discussions were audiotaped and transcribed. The transcripts were analyzed using qualitative methodology to determine relevant themes. RESULTS: Thirty-two physicians (16 male and 16 female) participated in the focus groups. The majority were in group practice (81%). The average number of years in practice was 11.75. An analysis of the focus group session identified two major clinical themes with subcategories: (1) physician issues (ie, identification, treatment); and (2) patient issues (ie, barriers to identification, symptom presentation). CONCLUSIONS: The focus groups served as an effective method to engage family physicians in isolating their own as well as their patients' difficulties in confronting this serious problem. PMID- 8426139 TI - Barriers to adherence to preventive services reminder letters: the patient's perspective. AB - BACKGROUND: Despite an emerging consensus as to which preventive services are appropriate, a minority of patients receive them. Although adherence to recommendations for some interventions has increased, research studies have shown that adherence rates can be further improved through a better understanding of patient attitudes and motivations regarding preventive services. METHODS: Using components of the Patient Path Model, this study examined the response to patient reminder letters for cholesterol screening sent to 1077 adult patients between August and October 1990. The research strategy incorporated both quantitative and qualitative methods, including a telephone survey and focus group interviews of nonresponders to the reminder letter. RESULTS: Three hundred seven patients were surveyed by telephone to ascertain their reasons for nonresponse. One hundred fifty-four (50.2%) did not recall receiving the reminder letter, 84 (27.4%) recalled receiving the letter but did not recall its content, and 69 (22.5%) recalled both receiving the letter and its content. No consistent reason for nonadherence emerged among the 69 nonresponders who recalled the reminder. Twenty seven of the nonresponders who did not recall receiving the cholesterol reminder participated in the focus groups. The participants stressed the importance of distinguishing the reminder letter from a bill, conveying a personally relevant message, and addressing logistical barriers to preventive services. CONCLUSIONS: Careful attention to the format and content of patient reminder letters is necessary to improve adherence to preventive services recommendations. PMID- 8426140 TI - Impact of the Americans with Disabilities Act on family physicians. AB - The Americans with Disabilities Act of 1990 prohibits discrimination against persons with disabilities in employment, government services, public accommodations, public transportation, and telecommunications. This article reviews the impact of the law on the practice of family physicians. Pre employment medical evaluations are prohibited by law, but medical evaluations may be performed after an offer of employment and before job assignment has been made. Employment may be conditional on results only if medical confidentiality is protected, and exclusionary criteria are job related, applied universally, and do not discriminate against individuals with disabilities. The law provides that persons with disabilities will have equal access to medical care, through prohibiting discrimination based on disability and through the design and construction of medical offices. The law requires physicians who are covered by the law to make reasonable accommodations so that qualified employees and applicants can perform the essential functions of a job. PMID- 8426141 TI - Narcolepsy. AB - Narcolepsy afflicts more than 200,000 Americans. In most cases the first symptom of the disease, excessive daytime sleepiness, develops during childhood or adolescence. This initial presentation is followed by cataplexy or other auxiliary symptoms several years later. Not infrequently, many years pass before the proper diagnosis of narcolepsy is made. Narcolepsy is a chronic lifelong disease without periods of remission. Excessive daytime sleepiness, inappropriate sleep attacks, and the pathognomonic symptom of cataplexy, are diagnostic of narcolepsy. Confirmation of the disease is made by a multiple sleep latency test. Although still not being used for diagnostic purposes, the association between narcolepsy and the human leukocyte group A (HLA) antigen DR2 is the strongest so far described for any disease. With the help of psychosocial support, therapeutic naps, and medications, the patient with narcolepsy may be able to lead a normal life. Methylphenidate and imipramine are the two most widely used drugs for the treatment of daytime somnolence and cataplexy, respectively. PMID- 8426143 TI - Post-traumatic stress disorder in a child following an automobile accident. AB - A 3-year-old girl was diagnosed with post-traumatic stress disorder (PTSD) following a minor automobile accident. The child presented with nightmares, violent play, and trauma-specific fears. The common symptoms of PTSD in childhood are reviewed, and the importance of careful history-taking and prompt referral to a mental health professional are discussed. PMID- 8426142 TI - Loop electrosurgical excisional procedure. AB - Loop electrosurgical excisional procedure, or LEEP, also known as loop diathermy treatment, loop excision of the transformation zone (LETZ), and large loop excision of the transformation zone (LLETZ), is a new technique for outpatient diagnosis and treatment of dysplastic cervical lesions. This procedure produces good specimens for cytologic evaluation, carries a low risk of affecting childbearing ability, and is likely to replace cryotherapy or laser treatment for cervical neoplasias. LEEP uses low-current, high-frequency electrical generators and thin stainless steel or tungsten loops to excise either lesions or the entire transformation zone. Complication rates are comparable to cryotherapy or laser treatment methods and include bleeding, incomplete removal of the lesion, and cervical stenosis. Compared with other methods, the advantages of LEEP include: removal of abnormal tissue in a manner permitting cytologic study, low cost, ease of acquiring necessary skills, and the ability to treat lesions with fewer visits. Patient acceptance of the procedure is high. Widespread use of LEEP by family physicians can be expected. PMID- 8426144 TI - Menometrorrhagia in an oral contraceptive user. AB - Endometrial carcinoma is the most frequent malignancy of the female reproductive tract, and irregular vaginal bleeding is the most common presenting symptom. Endometrial carcinoma is found most commonly among postmenopausal women and is associated with obesity, nulliparity, and anovulation. Oral contraceptive (OC) use and tobacco smoking have been reported to protect against endometrial carcinoma. Irregular vaginal bleeding is a common side effect of OC therapy. We report the case of an obese, premenopausal nulliparous woman with normal menses who developed menometrorrhagia and was then found to have endometrial carcinoma despite her youth and her use of both tobacco and combination OC. PMID- 8426145 TI - The fathers of percussion. PMID- 8426146 TI - Primary care and health care reform: the next 100 days. PMID- 8426147 TI - Disappearance rate of ethanol from the blood of human subjects: implications in forensic toxicology. AB - This article outlines major developments in knowledge about the human metabolism of ethanol. The results of a large number of controlled experiments aimed at measuring the rate of ethanol elimination from the blood are reported. The factors that influence the rate of ethanol elimination from blood, such as the amount of ethanol ingested, the drinking habits of the subjects, and the effect of food taken together with, or before, drinking were investigated. The slowest rate of ethanol disappearance was observed in a healthy male subject who ingested 0.68 g ethanol/kg body weight after an overnight (8 h) fast; the beta-slope was 9 mg/dL/h. The fastest rate of ethanol disappearance was observed in a male chronic alcoholic during detoxification; the beta-slope was 36 mg/dL/h. This four-fold difference in the rate of ethanol disposal should be considered when the pharmacokinetics of ethanol become an issue in drinking and driving trials, for example, when retrograde estimations are attempted. PMID- 8426148 TI - Nicotine analysis in neonates' hair for measuring gestational exposure to tobacco. AB - Hair samples were collected at time of delivery from 40 neonates whose mothers were known to be smokers during the prenatal period. Hair was decontaminated in dichloromethane, homogenized in NaOH, and nicotine was extracted in diethyl ether. After separation on a BP-5 capillary column, nicotine was identified and quantified by GC/MS using selected ion monitoring. In all cases, nicotine was found in the neonatal hair and in the hair of the corresponding mother. The ranges of nicotine levels were 0.15 to 11.80 ng/mg, and 0.37 to 63.50 ng/mg, for the neonates, and their mothers, respectively. It was possible to establish a significant correlation between both concentrations, and the correlation coefficient was 0.83. These findings suggest the possibility of monitoring the transfer of maternal nicotine through the placenta by measuring nicotine concentration in neonatal hair. PMID- 8426149 TI - The use of hydrogen peroxide to visualize tattoos obscured by decomposition and mummification. AB - Tattoos are distinctive, potentially important acquired identifying features that last for the life of the bearer, and persist into the postmortem period. However, the accumulation of decompositional pigments in the skin and subcutaneum will obscure tattoo designs. By treating the area with 3% hydrogen peroxide, the dark red-black decomposition products are temporarily removed, allowing underlying tattoos to be located, and photographed for identification purposes. PMID- 8426150 TI - A method for siding and sequencing human ribs. AB - Siding and sequencing (that is, putting in anatomical order) human ribs are essential to the proper examination and documentation of injuries to the chest. The paucity of information regarding sequencing nonfleshed human ribs makes it particularly difficult for physical anthropologists and medicolegal authorities to differentiate the midthoracic ribs. It was found that ribs could be accurately sequenced without the aid of a comparative skeleton using such features as maximum (relative) rib length, the size and shape of the articular facets, the distance between the articular facets and rib angle, and the height of the rib heads relative to one another. PMID- 8426151 TI - Evaluation of enzyme immunoassay performance characteristics--phencyclidine example. AB - Four reagent formulations (three provided by a manufacturer; one prepared in house by mixing equal volumes of two commercial reagents) are used for the assay of phencyclidine (PCP) in urine samples. Performance characteristics evaluated included assay precision and sensitivity at and near the assay cutoff concentration. Data resulting from the reagent prepared in-house are better than those using then commercially available formulations, and are comparable with those obtained using the recently available new commercial formulation. PMID- 8426152 TI - Child stealing by cesarean section: a psychiatric case report and review of the child stealing literature. AB - A highly unusual case of child stealing by cesarean section resulting in the death of the mother is presented. The judicial proceedings are summarized. The literature on the psychiatric status of perpetrators committing child stealing is presented. Two methods of psychiatrically categorizing these perpetrators are reviewed. The relevance of the literature to this and possible future cases is discussed. PMID- 8426153 TI - A technique for developing and photographing ridge impressions on decomposed water-soaked fingers. AB - One of the most challenging tasks confronting a crime laboratory technician is the fingerprinting and subsequent identification of an unknown homicide or drowning victim whose fingers have been subjected to a long period of exposure to water and the effects of decomposition. If the fingers of the individual have not been exposed to the erosive effects of water and decomposition for a long period of time, they may be allowed to dry, and suitable impressions are often obtainable. In other cases the fingers may have to be removed, with the permission of the Medical Examiners Office, and processed by the Crime Laboratory in an attempt to develop suitable ridge structure for inked impressions or an exact photographic copy of the individual's fingers. In extreme cases the effects of water and decomposition make the fragile ridge structure appear to be nonexistent to the naked eye. The procedure used in this case report, combines the use of cyanoacrylate vapor, commonly called "super glue fuming," and the ninhydrin process in conjunction to develop fragile ridge structure into discernable ridges that are easily seen and photographed for the purpose of making an identification of the individual. PMID- 8426154 TI - Scleral and conjunctival hemorrhages arising from a gunshot wound of the chest: a case report. AB - A young man committed suicide in the back seat of an automobile, witnessed by two law enforcement officers sitting in the front seat who had attempted to dissuade him from killing himself over an eight hour period. Death was caused by a contact gunshot wound of the anterior chest, which entered the midsternum and disrupted the anterior right atrium and ventricle of the heart, without involvement of either left ventricle or atrium. At the autopsy, bilateral fresh, confluent scleral and conjunctival sulcus hemorrhages were discovered, with no other evidence of facial or intracranial trauma. These hemorrhages are postulated to have arisen from a sudden pressure wave ascending through the superior vena cava, in a manner similar to the ocular findings associated with the retrograde venous blood flow that occurs during severe thoracic compression. These hemorrhages should not be mistaken for evidence that a decedent was beaten or otherwise involved in an assault episode. PMID- 8426155 TI - Sudden and unexpected death in childhood due to a colloid cyst of the third ventricle. AB - A nine-year-old boy died suddenly and unexpectedly following a two day history of intermittent headaches. At autopsy a colloid cyst of the third ventricle was found that had obstructed the foramen of Monro and caused hydrocephalus with prominent cerebral edema. Colloid cysts are rare entities in childhood and are not usually included in the differential diagnosis of pediatric sudden death. This report describes the clinicopathological features of such a case. PMID- 8426156 TI - The incidence of transient particulate gunshot primer residue in Oregon and Washington bow hunters. AB - The interpretation of GSR/PA (gunshot primer residue particulate analysis) results in an alleged firearm violation of bow hunting regulations is complicated by the theoretical presence of contaminant GSR from prior legitimate shooting incidents. A total of 120 samples representing field collections from thirty Oregon and Washington bow hunters were analyzed for the presence of particulate gunshot residue in order to assess the level of contamination that may be present in this population. Of the bow hunters sampled, 50% stated that they shoot guns; 80% of the shooting group stated at the time of the GSR field collection that they were wearing the same outer clothing or driving the same vehicle, or both, when they last handled and fired a weapon. Analysis of the 120 samples resulted in the detection of one tricomponent particle of GSR. PMID- 8426157 TI - Application of DNA fingerprinting to enforcement of hunting regulations in Ontario. AB - DNA fingerprinting has been used in investigations of 40 cases of infractions of hunting regulations involving white-tailed deer (Odocoileus virginianus) and moose (Alces alces) in Ontario. In most of these cases, individual-specific DNA fingerprints obtained with the Jeffrey's 33.15 multilocus probe were used to link the animal remains found at the illegal kill site to blood and tissue samples of the dead animal associated with a suspect. DNA fingerprints from 27 white-tailed deer and 19 moose were obtained in order to establish the level of band-sharing in DNA fingerprints among unrelated individuals in each species. We also determined the levels of band-sharing among animals from the same region and calculated the probability of two individuals sharing the same DNA fingerprint. Details are presented from cases in which the evidence was presented and accepted by Ontario courts. PMID- 8426158 TI - Extraction, evaluation, and amplification of DNA from decalcified and undecalcified United States Civil War bone. AB - Deoxyribonucleic acid (DNA) was extracted from documented skeletal specimens of U.S. Civil War soldiers to determine the need for decalcification prior to extraction. The polymerase chain reaction (PCR) was performed to determine if the calcification state had an effect on the ability to amplify the extracts and to determine how successful amplification would be with these aged specimens. Bone samples were pulverized to a fine powder and divided into two sets. One set of samples was decalcified and the other set left undecalcified. Both sets were extracted using an organic procedure. The results demonstrate that decalcification is not a necessary step in the extraction process and that the yield of DNA is generally two times greater when decalcification is omitted. Furthermore, the calcification state had no effect on the ability to perform the PCR. Although the extracted DNA was very degraded, a 410 base pair (bp) segment of the mitochondrial DNA (mtDNA) control region was amplified. These results suggest that DNA can be extracted and amplified from 125 year old bone without decalcification, which may assist in the identity of modern and historic forensic specimens. PMID- 8426159 TI - Problems of recovering partial human remains at different times and locations: concerns for death investigators. AB - Examples of cases where partial remains of the same individual were recovered at different times and from separate locations are presented. Such remains raise unique problems for coroner/medical examiners and police because their discovery has the potential to confound identification and disrupt investigative continuity. Recovered partial human remains highlight the need for their proper documentation and raise the question of their release for burial or retention for evidence. PMID- 8426160 TI - Poisoning from carbofuran. PMID- 8426161 TI - Discussion of H substance in urine. PMID- 8426162 TI - An animal model of postmortem amitriptyline redistribution. AB - An experimental rat model was developed to study postmortem changes of drug concentration after an acute overdose. Overnight fasted rats were fed 75 mg of amitriptyline (AMI). Two h after dosing, the rats were anaesthetized and blood samples were drawn from the femoral vein (peripheral blood--PB) and the heart (HB). The rats were sacrificed by CO2 and left at room temperature for either 0.1, 0.5, 1, 2, 5, 10, 24, 48, or 96 hours, when samples of heart blood, blood from the inferior vena cava (PB) and tissue samples from different liver lobes, heart, lungs, kidney, thigh muscle, and brain were taken. Samples were analyzed by high performance liquid chromatography. The AMI concentration in HB increased fairly rapidly within the first 2 h postmortem and from then the average ratio was 6.4 +/- 0.8 (mean +/- sem) (n = 31). In PB, the post/antemortem AMI concentration ratio followed an approximately exponential rise; at 2 h postmortem the ratio was 1.6 +/- 0.3 (n = 5), and at 96 h 55.1 +/- 23.8 (n = 4). For the main metabolite nortriptyline (NOR), the concentration changes followed the same pattern, but to a lesser extent. Among the tissues, the liver lobes had high, but variable drug concentrations; lobes lying closest to the stomach had the highest drug concentrations. The drug concentration in the lungs declined significantly. This animal model demonstrates postmortem drug concentration changes similar to those described in humans. Probable mechanisms include drug diffusion from the stomach and GI tract to the surrounding tissues and blood; and postmortem drug release from the lungs and possibly other drug-rich tissues into the blood. PMID- 8426163 TI - Experimental observations on adipocere formation. AB - Adipocere, "grave wax," is a waxy or greasy decomposition product formed by hydrolysis and hydrogenation of tissue fats. Once formed, it appears stable for extended periods. Adipocere has generally been considered to result from bacterial action, commonly in warm, damp, anaerobic environments. However, its frequency, rate of formation, factors affecting its formation and physical characteristics are not well defined. To study the frequency, time course, and effects of temperature and clothing on adipocere formation, we submerged human adipose tissue samples in aquaria under controlled conditions and conducted serial observations. Adipocere formed with high frequency, within a few months, in tissues submerged in warm tap water; similar changes took longer, 12 to 18 months in cold water submersion. Presence of clothing over the tissue appeared to accelerate adipocere formation. PMID- 8426164 TI - Invasive Haemophilus influenzae type B disease. AB - Invasive bacterial disease due to Haemophilus influenzae is a cause of sudden death in children. It must be considered by medical examiners when a child dies with a fulminant course and nonspecific symptoms. Three fatal cases are presented in children 7 weeks to 15 months of age. Two had meningitis and petechiae or purpura. All three had bilateral adrenal hemorrhage and a rapidly fatal course. The potential for rapid and accurate diagnosis of H. influenzae infection is widely available due to latex agglutination technique against bacterial capsular wall antigens. Diagnosis is critical because of its public-health implications. Up to 50% of cases may be acquired in day-care settings. Chemoprophylaxis is recommended for household and day care contacts. With the recent introduction of Haemophilus b conjugate vaccines for routine administration to infants beginning at 2 months of age, a change in the epidemiology of the disease is anticipated. PMID- 8426165 TI - The analysis of cremains: A case study involving the inappropriate disposal of mortuary remains. AB - Cremation as a method of body disposition has increased over the last two decades. Mishandling of the remains is bringing the analysis of cremains to the attention of forensic anthropologists. The destruction of skeletal components by heat, and mechanical reduction, make the analysis of cremains difficult. Three years after receiving cremains believed to be that of their mother, a family received a second set of cremains that were also purported to be those of their mother. Questioning whether which, if either, set of remains were the decedent, the family initiated an anthropological analysis of both sets of remains. The investigation focused on both osseous and nonosseous characteristics of each set. Total weight of the material and differences in both the osseous and nonosseous characteristics indicate that the cremains represent two individuals. These differences include the degree of color change due to heat, the amount of bone fragments vs. ash, and the lead and calcium content remaining in the fragments from each set. Differences in the nonosseous components of each set are related to coffin hardware, and medical and dental artifacts. PMID- 8426166 TI - Generalized anxiety disorder after stroke. AB - A series of 309 admissions to a stroke unit was examined for anxiety symptoms. Patients were diagnosed with DSM-III-R generalized anxiety disorder (GAD) symptom criteria. They were divided into groups of no anxiety (59.2%), worried but not fulfilling GAD criteria (13.9%), and GAD (26.9%). Patients were then divided into depressed and nondepressed groups based on the existence of DSM-III major or minor (dysthymic) depression. These groups were not significantly different in their background characteristics, family or personal psychiatric history, social support or the severity of physical impairment. Anxiety plus depression was associated with left cortical lesions, whereas anxiety alone was associated with right hemisphere lesions. Patients with worry had anterior and patients with GAD had posterior right hemisphere lesions. These findings suggest that anxiety disorder (independent of depression) is not related to background characteristics or to severity of impairment but is, in part, influenced by the brain structures that are injured. PMID- 8426167 TI - Neuropsychological assessment of attention problems in pathological gamblers. AB - Deficits in executive, frontally mediated attention processes have been observed in substance abusers. A significant rate of childhood histories of attention deficit hyperactivity disorder has also been reported in this population, creating the question of whether attention problems predated addiction or were secondary to neurological drug or alcohol effects in pathological gamblers. To address this issue, the current study compared 33 non-substance-abusing pathological gamblers with 33 nonaddicted controls on nine attention measures and childhood behavior questionnaires. Gamblers performed significantly worse than controls on higher order attention measures and reported more childhood behaviors consistent with attention deficits. Results suggest attention deficits may be a risk factor for development of addictive disorders. PMID- 8426168 TI - DSM-III-R disorders in Vietnamese refugees. Prevalence and correlates. AB - This study's purpose was a) to determine the prevalence of DSM-III-R disorders in newly arrived ethnic Vietnamese and ethnic Chinese refugees from Vietnam and b) to determine the correlates of DSM-III-R disorders. A Vietnamese-speaking psychiatrist administered translated sections of the Structured Clinical Interview for DSM-III-R to 201 Vietnamese new arrivals undergoing mandatory health screening. Overall, 18.4% had one or more current disorders: 8.5% had adjustment disorder and 5.5% had major depression. Ethnic Vietnamese, compared with ethnic Chinese, had significantly (p < .05) higher rates of current posttraumatic stress disorder and generalized anxiety disorder. Ethnic differences in psychopathology were largely explained by the fact that ethnic Vietnamese refugees had experienced more traumatic events and separation from family. After adjusting for ethnicity, refugees who reported traumatic events, refugees who were married, and veterans were significantly (p < .05) more likely to have one or more psychiatric disorders. PMID- 8426169 TI - Sociodemographic and disease-related correlates of depressive morbidity among diabetic patients in Zagreb, Croatia. AB - The magnitude of depressive morbidity as well as its significant sociodemographic and disease-related correlates were investigated in a sample of 180 adult diabetic patients attending an outpatient clinic in Zagreb, Croatia (formerly Yugoslavia) in 1989. Results using the Zung Self-Rating Depression Scale yielded a prevalence rate of 60.5% with 10% manifesting severe levels of depressive symptoms. Significant correlates of higher symptoms included older age, female gender, unmarried status, less education, the presence of diabetic complications, longer duration of the disease, more demanding diabetic regimens, and poorer adherence to the regimen. The lowest coefficient was for level of glycosylated hemoglobin (r = .04), indicating a weak relationship between diabetic control and depressive symptoms. Findings are discussed in relation to other epidemiological surveys of depression and to the deteriorating economic and political situation in Croatia. The implications of untreated depression for diabetic self-care are discussed. PMID- 8426170 TI - A method to detect intercenter differences in the application of contemporary diagnostic criteria. AB - The use of specified diagnostic criteria is part of routine psychiatric practice and research. However, there is evidence that the same criteria may be interpreted and applied differently at different research centers. The next question is whether it is possible to detect these systematic intercenter differences in diagnostic practices. An inexpensive, nonlaborious, standardized method is needed that can easily be used at any site. In the present report, we demonstrate how self-report questionnaires can provide a method of detecting systematic differences in the application of contemporary diagnostic criteria. Sixty consecutively admitted inpatients with nonpsychotic major depressive disorder were interviewed with standardized schedules and diagnosed according to two interpretations of the Research Diagnostic Criteria (RDC) and DSM-III endogenous/melancholia criteria. The patients also completed the Inventory to Diagnose Depression, a self-report scale designed to diagnose major depression and subtype patients according to RDC and DSM-III endogenous/melancholia criteria. The interview to self-report ratio for diagnosing endogenous depression and melancholia was associated with diagnosticians' interpretation of the criteria. A broader application of the criteria resulted in a higher ratio. A paper-and-pencil questionnaire could be useful in detecting systematic interinvestigator differences in the broadness or narrowness with which criteria are applied. The advantage of a self-report scale is that it is free from systematic biases and tendencies of clinician raters. Of course, ratings on questionnaires are not free of bias; however, it is less likely that differences between samples will reflect a systematic variation in response set. PMID- 8426171 TI - Obstetric factitious disorder and Munchausen syndrome by proxy. PMID- 8426172 TI - Comment on spontaneous seizure activity. PMID- 8426173 TI - Spontaneous seizure attributable to prolonged seizure, not unilateral electroconvulsive therapy. PMID- 8426174 TI - One-year follow-up status of treatment-seeking cocaine abusers. Psychopathology and dependence severity as predictors of outcome. AB - While the prognostic significance of comorbid psychopathology and dependence severity has been demonstrated with opiate addicts and alcoholics, no previous reports have examined these issues in cocaine abusers. We reinterviewed 94 cocaine abusers 1 year after they sought treatment to assess predictors of treatment retention and outcome. Results suggested that: a) Many cocaine abusers did comparatively well; fully a third reported complete abstinence during the 12 months preceding the follow-up interview. b) Three variables emerged as consistent predictors across several outcome domains: severity of drug use, poorer psychiatric functioning, and presence of concurrent alcoholism. c) The data supported a unidimensional model of outcome for cocaine abuse that emphasized reduction in level of substance use. However, abstinence was not strongly associated with improved functioning in all outcome areas. d) Variables associated with longer retention in treatment also tended to predict poorer outcome. PMID- 8426175 TI - Interpersonal stressors, substance abuse, and suicide. AB - In contrast to suicide in depression, suicide associated with alcoholism and substance dependence may be preceded more often by interpersonal loss and conflict 6 weeks prior to death. We used psychological autopsy methodology in an effort to clarify and extend these findings using a more comprehensive typology of interpersonal stressors. Subjects included 57 suicide victims with diagnoses of alcohol/substance dependence (A/SD; N = 30) and mood/anxiety disorders (M/AD; N = 27). Consistent with previous studies, a substantial majority of the A/SD suicide victims were confronted with interpersonal stressors in the 6 weeks prior to death. Our investigation extends previous findings by indicating that a) A/SD suicide victims are confronted with a broader range of interpersonal stressors than M/AD suicide completers and b) the types of interpersonal stressors experienced by A/SD subjects in the weeks prior to suicide involve conflicts/arguments and attachment disruptions. PMID- 8426176 TI - Rehabilitation for adults with severe mental illness and substance use disorders. A clinical trial. AB - Faced with many patients with comorbid severe mental illness and substance use disorders, a university-affiliated, inner-city community mental health center and a psychosocial rehabilitation center initiated this clinical trial assessing a program to care for these patients. Fifty-four patients, age 18 to 40, with either schizophrenia or major affective disorder and a substance use disorder were randomly assigned to usual community mental health center and rehabilitation services with or without an innovative group and intensive case management program. One-year follow-ups detected no significant advantages on patient outcomes for adding the innovative program to usual services. Failure to engage patients in the experimental program posed a major and enduring barrier to treatment, despite intensive case management. Future efforts must give greater consideration to effective engagement techniques and patients' readiness for active treatment. PMID- 8426177 TI - Are there symptoms that are specific for depressed mood in patients with traumatic brain injury? AB - This study examined the specificity of vegetative and psychological symptoms of depression among 66 patients with acute traumatic brain injury followed over 1 year. The median frequencies of vegetative and psychological symptoms among patients with depressed mood were 3 and 3. These frequencies were three times the respective rates among nondepressed patients. Although change in self-attitude and subjective anergia distinguished depressed from nondepressed patients throughout the 1-year follow-up, some symptoms, such as early awakening and difficulty concentrating, distinguished groups only after 6 months. If diagnostic criteria for major depression were modified to include only specific symptoms of depression, the standard (i.e., unmodified) DSM-III-R still had a 100% sensitivity and 94% specificity at the initial evaluation and 80% and 100%, respectively, at 1 year. There were almost no patients with depressive symptoms without a depressed mood (i.e., "masked" depressions). These findings suggest that DSM-III-R criteria for major depression are useful, even in an acute head injured population, but also suggest that the nature of posttraumatic brain injury depressive disorder may change over time. PMID- 8426178 TI - Spinal cord injury and vitamin D metabolism. PMID- 8426179 TI - The development of urologic complications in relationship to bladder pressure in spinal cord injured patients. AB - The medical records of 88 patients followed through our spinal cord injury clinic were reviewed to determine if elevated intravesical pressures result in more urologic complications than are seen with low pressure bladders. Fifty-two of the patients were noted to have a high bladder pressure (sustained detrusor pressure greater than 40 cm water) on cystogram while 36 had low pressures. All patients had routine urine cultures, urodynamics, ultrasonography, radioisotope renal scans, and excretory urograms. Bladder management was directed at maintaining a low bladder pressure and included one or more of the following: intermittent catheterization, anticholinergics, alpha blockers, transurethral sphincterotomy, or indwelling catheters. Average follow-up was 6 years. Mild degrees of hydronephrosis were noted in seven (14 percent) of the patients with a high pressure bladder and in one (3 percent) with a low pressure bladder. Pyelonephritis was noted in two (4 percent) with high bladder pressure and two (5 percent) with low bladder pressure. Preservation of renal function occurred as the result of patient compliance with bladder management and bladder pressure. Sustained high detrusor pressure, when not corrected, leads to upper tract deterioration which was reversed by aggressive lower tract management. PMID- 8426180 TI - Bladder neck closure with continent augmentation or suprapubic catheter in patients with neurogenic bladders. AB - Twenty-four patients, with various combinations of non-healing decubitus ulcers, urethral fistulae, incontinence, and penile skin breakdown were candidates for proximal urinary diversion, having failed intermittent, external, and indwelling catheterization combined with pharmacologic therapy. Seventeen patients underwent bladder neck closure, including seven with multiple sclerosis and ten with spinal cord injury, and because they were unable or unwilling to do catheterization, had their urine diverted by suprapubic catheter. Seven patients, including four with spinal cord injury, underwent bladder neck closure and continent augmentation with formation of a catheterizable cutaneous stoma on the anterior abdominal wall, using right colon and right colon/ileum configurations. When ureteral reflux and obstruction are absent, the patient's bladder was used which spared the added risk of ureteral implantation and possible ureteral stricture while increasing total bladder capacity. In a select group of patients with intractable incontinence, perineal and penile skin breakdown, or urethral fistulae, bladder neck closure and urinary diversion by suprapubic catheter or continent augmentation has proven to be a reliable and effective alternative to an ileal conduit. PMID- 8426181 TI - The effects of multidisciplinary team care for acute spinal cord injury patients. AB - The care of 169 survivors of spinal injury receiving acute treatment at the Hamilton General Hospital in Ontario, Canada, was studied. This paper compares treatment of groups of patients before and after formation of a multidisciplinary Acute Spinal Cord Injury Team. The establishment of multidisciplinary team care for acute spinal cord injury patients in our tertiary referral center correlates with clinically and statistically significant reductions in length of stay in the acute care hospital, alterations in the rate of surgical treatment for them, changes in the use of radiological resources, and reduction in the average number of days febrile. The team also brought stronger representation of allied health professionals to the hospital records of acute spinal cord injury patients. These important changes result from implementation of an effective multidisciplinary medical team without the addition of new funds, personnel, or hospital facilities and without alteration in referral patterns. Our team did not reduce mortality, duration of intensive care unit stay, or work for physicians. PMID- 8426182 TI - Circulating levels of soluble interleukin 2 receptors are elevated in the sera of humans with spinal cord injury. AB - A unique molecular regulatory mechanism or final common molecular pathway mediating the autonomic dysfunction and several pathobiologic sequelae of spinal cord injury (SCI) in humans has not been delineated. Although seemingly disparate in etiopathogenesis, much of the pathology caused by traumatic disruption of the spinal cord may be attributable to the pleiotropism demonstrated by a unique family of endogenous bioactive molecules, the interleukins. To begin testing this hypothesis, we examined the sera of patients with chronic SCI for elevations in interleukin 1 beta (IL-1 beta) and interleukin 2 receptor (IL-2R) and compared them to a control population of able-bodied subjects. In comparison to control subjects, a statistically significant increase in IL-2R was observed in patients with cervical spinal myelopathy. Elevated levels of IL-2R were not seen in paraplegic patients. Significant differences between the means and variances of serum IL-1 beta could not be detected among the study groups. We conclude that the sera of quadriplegic patients with chronic SCI contain elevated levels of IL 2R and suggest that the elevated levels of IL-2R may be of diagnostic, prognostic, and therapeutic importance. PMID- 8426183 TI - The VA responsibility in tomorrow's national health care system. PMID- 8426184 TI - Effects of chronic spinal cord injury and pressure ulcer on 25(OH)-vitamin D levels. AB - We studied 92 spinal cord injured (SCI) men (50 paraplegics and 42 quadriplegics) with normal renal function, 38 of whom had single or multiple pressure ulcers. The results were compared with those of 28 able-bodied normal controls. Serum concentrations of calcium and magnesium were measured by atomic absorption spectrometry, and 25(OH)-vitamin D was quantitated by a specific competitive binding assay using a sensitive vitamin D binding protein and tritiated 25(OH) vitamin D. The SCI group exhibited significant reductions in serum 25(OH)-vitamin D and total calcium concentrations as compared to the normal control group. Although the mean serum concentration of 25(OH)-vitamin D in the quadriplegic patients as a whole was lower than that found in the entire paraplegic group, the difference did not attain statistical significance. Similar observations were made when the ulcer-free subgroups of paraplegics and quadriplegics were compared. The SCI subgroup which was least physically active, i.e., those with pressure ulcers, showed the greatest depression of serum 25(OH)-vitamin D, calcium, and magnesium concentrations. The observed reduction in serum 25(OH) vitamin D in SCI patients appears to be partly related to reduced cutaneous vitamin D biosynthesis from sunlight deprivation occasioned by physical disability and hospitalization. In addition, nutritional deficiency and altered intestinal transport may be involved. The reduction in serum calcium concentration may be related to abnormal vitamin D metabolism and hypoalbuminemia (reduced protein-bound calcium). PMID- 8426185 TI - Neuropathology of mitochondrial encephalomyopathies due to mitochondrial DNA defects. PMID- 8426186 TI - The importance of fibrinoid necrosis as the cause of cerebral hemorrhage in hypertension. Commentary. PMID- 8426187 TI - Anatomic distribution of the growth-associated protein GAP-43 in the developing human brainstem. AB - GAP-43 is a membrane phosphoprotein whose expression is high in neurons undergoing development or remodeling of axonal connections. This study used a monospecific antibody to GAP-43 to investigate the sequences of fiber tract elongation and synaptic development in the human brainstem. Immunocytochemistry was performed in 14 fetal and infant brainstems; two child and adult cases were also examined for comparison. At midgestation, GAP-43 immunostaining was moderately intense across nuclei and fiber tracts, except for the corticospinal tract, where levels were higher, and cranial nerve nucleus VII, superior olive, inferior colliculus, inferior olivary hilum, inferior cerebellar peduncle, medial lemniscus, and medial longitudinal fasciculus, where staining was nearly absent. By the end of the neonatal period, the relative distribution of GAP-43 immunostaining appeared well-established and similar, although not identical, to that in the child and adult brainstem. Immunostaining was absent or negligible in almost all the cranial nerve somato- and branchiomotor nuclei, auditory-relay nuclei, and vestibular nuclei, while remaining intense in visceral-related nuclei, reticular formation, cochlear nucleus, and periaqueductal gray. Staining was also virtually absent in all fiber tracts at birth, except for the corticospinal tract and central tegmental tract. Persistence of GAP-43 staining in the corticospinal tract past the fetal period suggests that this tract remains in a plastic state beyond initial axonal elongation. Intense immunostaining in visceral-related nuclei into adulthood suggests that these regions may continue to undergo synaptic reorganization. This study provides baseline information relevant to understanding developmental brainstem disorders in early human life. PMID- 8426188 TI - Central nervous system malformations in trisomy 9. AB - Trisomy 21, 18 and 13 are the most common varieties of autosomal trisomy recognized at birth; most of the others lead to spontaneous abortions in the first trimester. Trisomy 9, a rare trisomy, is compatible with life, but, unlike trisomy 21, 18 and 13, the range of manifestations has not been well catalogued. Central nervous system abnormalities have been reported in the majority of cases, usually including a dilated fourth ventricle and malformed cerebellum. The posterior fossa malformation closely resembles the descriptions of the Dandy Walker malformation leading some to suggest this designation, while others have suggested that the features are unique to trisomy 9. Two cases of trisomy 9 are presented in this report which extend the range of neuropathologic manifestations in this cytogenetic disorder. The first infant had cortical migration abnormalities, anomalous hippocampal formation, simplified inferior olivary nuclei, germinal matrix cysts, mild ventriculomegaly, syringomyelia, and a large myelomeningocele without a Chiari type II malformation. The fourth ventricle was normal in size and the cerebellum unremarkable. The second infant had a cystically dilated fourth ventricle and widely separated cerebellar hemispheres with an intact cerebellar vermis, the features of which we felt were compatible with the Dandy-Walker malformation. In addition, agenesis of the corpus callosum, anomalous hippocampal formation, subpial glial nodules and mild ventriculomegaly were present. These cases extend the range of malformations that may be associated with trisomy 9, and raise the differential diagnosis of trisomy 9 when these malformations are identified.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426189 TI - Schwann cell injury is attenuated by aldose reductase inhibition in galactose intoxication. AB - Four months of galactose intoxication induces a dose-dependent osmotic imbalance of the nerve microenvironment characterized by polyol, water, and electrolyte accumulation. Recently, dose-dependent cellular lesions have been described in the sciatic nerves of galactose-intoxicated rats. The present study was designed to demonstrate that the cell injury and endoneurial osmotic imbalance in galactose intoxication are dependent on the subsequent metabolism of galactose by the polyol pathway. Three groups of age-matched, female Sprague-Dawley rats were fed a control diet or diets containing complete micronutrient supplements with 40% galactose or 40% galactose and 0.04% Ponalrestat, an aldose reductase inhibitor (ARI). After 4 to 5 months, sciatic nerves were analyzed for polyol, water and endoneurial electrolyte content and processed for light and electron microscopic examination. Ponalrestat prevented myo-inositol depletion and accumulations of dulcitol, water and endoneurial fluid electrolytes. Axonal size frequency histograms revealed that Ponalrestat attenuated the shift toward smaller fibers and the decrease in mean axonal diameter seen in untreated galactose-fed rats. Electron microscopic examination showed widespread reactive and degenerative changes in Schwann cells of galactose-intoxicated rats that culminated in cytoplasmic disintegration. Quantitative electron microscopy revealed that ARI treatment significantly reduced the incidence of abnormal Schwann cells. These observations indicate that the osmotic imbalance and cell injury seen in galactose intoxication are dependent on the metabolism of galactose by the polyol pathway. PMID- 8426190 TI - An immunogold electron microscopic analysis of FMRFamide-like immunoreactive neurons in the CNS of Helix pomatia: ultrastructure and synaptic connections. AB - The ultrastructure and synaptic connections of FMRFamide-like immunoreactive neurons were investigated in the CNS of the snail Helix pomatia, following the application of a post-embedding immunogold method. For comparison, first, we analyzed the ultrastructure and granule content of the identified FMRFamide containing C3 neuron in the cerebral ganglion. Three types of unidentified immunoreactive neuronal perikarya, and five types of varicosities could be distinguished on the basis of granule content. The different granule types revealed a highly selective accumulation of gold particles. One granule type contained by one FMRFamide-like immunoreactive neuron type (N1) and by one varicosity type (T2) showed similar ultrastructure to that of the granules seen in the C3 cell. In the neuropil, the majority of FMRFamide-like immunoreactive varicosities (four of the five varicosity types) established specialized synaptic contacts with unidentified postsynaptic profiles. In the connective tissue sheath around the ganglia, three types of FMRFamide-like immunoreactive varicosities were found to establish unspecialized contacts with smooth muscle fibres or to be free in the mass of collagen fibres. On the basis of these observations, we suggest (1) an extensive diversity of the localization of FMRFamide (and related substances) at the ultrastructural level; (2) the involvement of FMRFamide-like immunoreactive varicosities in synaptic, modulatory and neurohormonal regulatory processes in the Helix nervous system. PMID- 8426191 TI - Protein gene product 9.5 in the developing cochlea of the rat: cellular distribution and relation to the cochlear cytoskeleton. AB - Protein gene product 9.5 was immunolocalized in the adult and early postnatal (P2 P15) rat cochlea, and its distribution compared with a 200 kDa highly phosphorylated neurofilament subunit (neurofilament 200) and alpha-tubulin. In the adult, Protein gene product 9.5 was expressed exclusively in cochlear nerve fibres and ganglion cells, a small percentage of these (Type II ganglion cells and olivocochlear bundle fibres) being intensely positive for both protein gene product and neurofilament 200. In postnatal development, pillar and Deiters' cells were at first (P2-P15) strongly positive for protein gene product 9.5, and hair cells moderately so. At P2, all nerve fibres and ganglion cells showed co expression of protein gene product 9.5 and neurofilament 200, but at later stages, the subset of intensely co-labelled neurons appeared, nerve fibres at P7 onwards and ganglion cells from P12. There was no overt correlation between the onset of protein gene product 9.5 and alpha-tubulin expression in any cochlear component. Protein gene product 9.5 expression in ganglion cells was at first (P2 and P7) mainly nuclear, and later also cytoplasmic. It is concluded that there is a clear correlation of high levels of protein gene product 9.5 and neurofilament protein expression, and that protein gene product 9.5 is expressed in some non neuronal cells of the cochlea during its early development, persisting until after hearing has commenced. PMID- 8426192 TI - The distribution of GAP-43 in normal rat spinal cord. AB - We have studied the distribution of the growth-associated protein GAP-43 in the spinal cord of adult rats by light and electron microscopy, using a new antiserum raised against GAP-43/beta-galactosidase fusion protein. We show that GAP-43 is present at all vertebral levels but is more concentrated in cervical and thoracic regions. In addition to heavy staining in the corticospinal tracts of the white matter, staining can be seen at the light microscopic level throughout the grey matter and is particularly heavy around the central canal and in the superficial dorsal horn. Electron microscopic examination revealed that GAP-43 immunostaining is confined to a subpopulation of axons and axon terminals. Staining occurs in small myelinated and unmyelinated fibres and in terminals which are mainly small and make single axodendritic or axosomatic synapses. Staining in such terminals occurs in the axoplasm but is heaviest immediately adjoining the axolema. Staining was not observed in dendrites, nor in large myelinated axons or large axon terminals. Our results indicate that GAP-43 is expressed in adult rat spinal cord in a subpopulation of small diameter fibres and axon terminals. The distribution and morphology of these terminals is consistent with several different possible origins including corticospinal projection neurons, small diameter primary afferent neurons, and descending raphe-spinal serotonin containing neurons. PMID- 8426193 TI - Up-regulation of GAP-43 and growth of axons in rat spinal cord after compression injury. AB - The growth-associated protein-43 (GAP-43) is an axonal phosphoprotein which is expressed at high levels during development and is reinduced by regeneration in the PNS. Consequently it is believed to be a key molecule in the regulation of axonal growth. However, injury to the CNS does not result in significant regeneration and this has been suggested to correlate with a failure of central neurons to up-regulate GAP-43 after axotomy. We have examined a model of spinal cord injury which is unique in two respects; first dural integrity is maintained by compression of the cord with smooth forceps (thus excluding connective tissue elements) and, secondly, considerable axonal growth has been reported through the resulting lesion. Our previous studies have shown that GAP-43 is extensively distributed in the rat spinal cord (see accompanying paper), but here we have used anti-GAP-43 antiserum at a dilution which did not yield any immunostaining in normal cord. However, supranormal levels of GAP-43 were detected in cell bodies and axons around the lesion within four days of compression injury. Double immunostaining with the RT97 monoclonal antibody indicated that a small subpopulation of neurons local to the site of compression were axotomized and expressed GAP-43 and phosphorylated neurofilament epitopes in their cell bodies. Although damage to long axon tracts was extensive, there was no evidence of regeneration in white matter. On the other hand cavities which formed in grey matter provided an environment for axonal elongation. Immunolabelling with markers for astrocytes and endothelial cells was used to evaluate the interaction of elongating axons with endogenous CNS cell types. Sprouting axons, identified by the presence of elevated levels of GAP-43, did not appear to grow in contact with astrocytes but preliminary evidence suggested that newly formed capillaries provided an appropriate substrate. PMID- 8426194 TI - Secondary acute myeloid leukemia following treatment with epipodophyllotoxins. PMID- 8426195 TI - What's the deal with follicular lymphomas? PMID- 8426196 TI - Secondary acute myeloid leukemia in children with acute lymphoblastic leukemia treated with etoposide. AB - PURPOSE: To describe the occurrence of secondary acute myeloid leukemia (AML) in children with acute lymphoblastic leukemia (ALL) treated with etoposide (VP-16). PATIENTS AND METHODS: Two hundred five consecutive children with early B-lineage ALL were treated according to the Dallas/Fort Worth (DFW) protocol between January 1986 and July 1, 1991. Therapy included a four-drug induction followed by consolidation and continuation phases of nightly oral mercaptopurine (6-MP) and repetitive courses of divided-dose oral methotrexate (dMTX) and asparaginase (L asp). Three doses of VP-16 and cytarabine (Ara-C) were given during consolidation and later, during continuation, two doses were given 3 to 4 days apart, every 9 weeks. Intrathecal (IT) chemotherapy was given throughout the treatment period. RESULTS: Two hundred three of the 205 patients entered remission. Only eight of these 203 children have had a bone marrow relapse (ALL). However, 10 other children have developed secondary AML 23 to 68 months following the diagnosis of ALL. Overall event-free survival (EFS) at 4 years is 79.3% +/- 5.1%, with a risk of secondary AML at 4 years of 5.9% +/- 3.2%. CONCLUSION: This experience provides strong evidence for a link between epipodophyllotoxin therapy and secondary AML since none of these children received alkylating agent therapy or irradiation. This serious complication raises concern as to the appropriate use of epipodophyllotoxins in the treatment of childhood ALL. PMID- 8426197 TI - Clinical features and prognosis of follicular large-cell lymphoma: a report from the Nebraska Lymphoma Study Group. AB - PURPOSE: Our purpose was to describe the treatment outcome of patients with follicular large-cell lymphoma (FLCL) and to identify prognostic factors that affect the treatment outcome. PATIENTS AND METHODS: Between 1980 and 1991, 107 newly diagnosed, previously untreated patients with FLCL were prospectively treated using treatment plans of the Nebraska Lymphoma Study Group (NLSG). Most stage I/II patients received two to three cycles of one of four closely related six-drug combination chemotherapy regimens (cyclophosphamide, doxorubicin or mitoxantrone, and procarbazine, plus bleomycin, vincristine, and prednisone or dexamethasone [CAP/BOP I-IV]) plus involved-field radiotherapy; 10 patients received involved-field irradiation only. Stage III/IV patients received six to eight cycles of CAP/BOP. RESULTS: Forty-four percent of patients had stage I/II disease. Stage I/II patients were older and more often female than stage III/IV patients. Cytogenetic studies were available on 35 patients: seven were normal; the most common abnormality was a translocation involving 14q32. Abnormalities of 1p or 1q were also common, often secondary to a 14q32 abnormality. The median follow-up of surviving patients is 2 years. The complete response rates observed were stage I/II, 88%; stage III/IV, 49%. Complete response rates were affected by both age and tumor bulk. Failure-free survival (FFS; time to first occurrence of progression, relapse after response, or death from any cause) at 3 years was estimated to be 61% for stage I/II patients and 34% for stage III/IV patients. Survival at 3 years was estimated to be 76% and 61%, respectively. FFS of stage III/IV patients was poorer for stage IV patients and those with composite lymphomas. Significantly poorer survival was only seen in patients older than 70 years of age. CONCLUSION: A proportion of stage I/II FLCL patients may obtain long-term disease control with combination chemotherapy plus radiotherapy. Results for patients with stage III/IV FLCL are similar to those seen for other follicular lymphomas. PMID- 8426198 TI - Late relapse in early-stage Hodgkin's disease patients enrolled on European Organization for Research and Treatment of Cancer protocols. AB - PURPOSE: The great majority of relapses after the treatment for early-stage Hodgkin's disease are observed within 4 to 5 years after treatment completion. This study describes the characteristics and outcome of patients who had late relapses, which was defined as relapses that occurred 5 or more years after initial treatment start. PATIENTS AND METHODS: A total of 1,082 adult patients with early clinical stage Hodgkin's disease were enrolled on three consecutive European Organization for Research and Treatment of Cancer (EORTC) protocols (H1, H2, and H5 trials) from 1964 to 1981. Of these, 1,044 patients satisfied the eligibility criteria with a supradiaphragmatic localization, age greater than 15 years, and initial complete remission. Overall, 341 patients (32.6%) relapsed, 304 (29.1%) early and 37 (3.5%) late. For each of these 37 late relapsers, questionnaires were sent to the participating centers and detailed information for 34 relapses was obtained. Cumulative probabilities for developing a late relapse were estimated using the Kaplan and Meier method. Quantification of the relationship between late relapse and several confounding variables was performed using the Cox's proportional hazards model. RESULTS: The 10- and 15-year cumulative probabilities of late relapse in patients who were disease-free at 5 years were 4.8% and 8.3%, respectively. Patients treated on more recent protocols had a higher incidence of late relapse, possibly due to an attempt to tailor therapy to the specific prognostic factors (10-year cumulative probabilities, 4.6%, 2.6%, and 7.5% in trials H1, H2, and H5, respectively). Incidence of late relapses significantly correlated with male sex, B symptoms, mediastinal involvement, and treatment modality. Salvage treatment induced a complete response in 27 patients (79%) and a prolonged complete remission in 24 patients (71%). Twenty years after initial treatment start, similar overall survival rates were observed for late relapsing (72%) and nonrelapsing patients (75%). CONCLUSION: Late relapses of Hodgkin's disease are uncommon, but may be more frequent with recent protocols tailored to specific prognostic factors. If treated, their outcome is favorable. Late relapse is therefore another factor indicating that careful, long-term follow-up is needed for patients with Hodgkin's disease. PMID- 8426199 TI - Central lymphatic irradiation for stage III nodular malignant lymphoma: long-term results. AB - PURPOSE: To report the long-term results of central lymphatic irradiation for stage III nodular malignant lymphoma. PATIENTS AND METHODS: Between 1969 and 1985, 34 patients (26 with nodular poorly differentiated lymphoma, four with nodular mixed lymphocytic/histiocytic lymphoma, and four with nodular histiocytic lymphoma) were treated with central lymphatic irradiation. Median age of the group was 51 years (range, 30 to 73). There were 15 men and 19 women. Staging work-up included a physical examination and bone marrow biopsy in all patients. Seventy-four percent had a lymphangiogram (LAG) and 44% a laparotomy (LAP). Eighty-two percent had either a LAP or a LAG. Thirty-two patients were Ann Arbor stage IIIA and two were stage IIIB. All patients received lymphatic irradiation that encompassed cervical, supraclavicular, axillary, mediastinal, paraaortic, mesenteric, pelvic, and femoral lymphatics to total doses ranging from 20 to 30 Gy in 1.0- to 1.8-Gy fractions. Waldeyer's ring was initially treated in 17 patients. Follow-up information is available on all 34 patients. Median follow-up is 9 years, 8 months (range, 15 to 244 months). RESULTS: Life-table actuarial overall, disease-free, and cause-specific survival rates at 15 years are 28%, 40%, and 46%, respectively. Only one relapse was observed after 9 years. Disease free survival was significantly improved in patients with five or fewer sites of involvement (P = .02). Age, sex, B symptoms, histology, and technique of irradiation were not prognostically significant. Salvage therapy, including further irradiation and/or chemotherapy, was delivered to 20 patients. Ten percent of these patients remain alive without evidence of disease. Toxicity data were available for the patients treated at the Medical College of Wisconsin (MCW). Radiation Therapy Oncology Group (RTOG) acute hematologic, gastrointestinal, and salivary toxicity scores were < or = 2 in 83% of patients. Late toxicity scores were < or = 2 in 96%. Persistent xerostomia was noted in 23% of patients who received initial treatment to Waldeyer's ring. CONCLUSION: These results suggest that initial comprehensive central lymphatic irradiation may be the preferred approach to achieve a durable relapse-free interval for this group of patients. PMID- 8426200 TI - Long-term reduction in sperm count after chemotherapy with and without radiation therapy for non-Hodgkin's lymphomas. AB - PURPOSE: Treatment of lymphomas with combination chemotherapy with or without radiation therapy (XRT) can result in long-term or permanent azoospermia. PATIENTS AND METHODS: Semen analyses of lymphoma patients were performed before, during, and after treatment with cyclophosphamide, doxorubicin, vincristine, prednisone, and bleomycin (CHOP-Bleo) chemotherapy. Some of the patients also received other drugs or radiation therapy. RESULTS: Although no patients were azoospermic before treatment, all were rendered azoospermic during treatment. Following the completion of treatment, the fraction of patients whose sperm counts recovered increased gradually over 5 years and plateaued by 7 years, with two thirds of the men achieving normospermic levels. Scattered gonadal radiation dose and cumulative cyclophosphamide dose were found to be independently significant determinants of recovery: the fraction of patients whose sperm counts recovered to 10 x 10(6)/mL were 83%, 47%, and 20% for those who received less than 9.5 g/m2 of cyclophosphamide, greater than 9.5 g/m2 of cyclophosphamide, and pelvic XRT, respectively. The inclusion of additional drugs and interferon alfa did not significantly affect the long-term recovery of spermatogenesis. CONCLUSION: Pelvic XRT and cumulative cyclophosphamide dosages greater than 9.5 g/m2 are associated with a high risk of permanent sterility in lymphoma patients treated with the CHOP-Bleo regimen. PMID- 8426201 TI - AMOPLACE treatment of intermediate-grade and high-grade malignant lymphoma: a Cancer and Leukemia Group B study. AB - PURPOSE: In an attempt to improve the efficacy of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy for intermediate-grade and high grade non-Hodgkin's lymphomas, a phase II evaluation of a regimen consisting of Adriamycin (doxorubicin; Adria Laboratories, Columbus, OH), methotrexate, Oncovin (vincristine; Eli Lilly Co, Indianapolis, IN), prednisone, leucovorin, cytarabine (ara-c), cyclophosphamide, and etoposide (AMOPLACE) was conducted. This regimen includes three additional agents not found in CHOP, uses weekly doses of alternating myelosuppressive and nonmyelosuppressive drugs, and incorporates most single agents active against diffuse lymphomas. PATIENTS AND METHODS: Ninety-one previously untreated patients were enrolled and 60 patients were confirmed eligible after central pathology review. Fifty-eight percent of patients had diffuse large-cell lymphoma (DLCL), 83% had stage III or IV disease, and 45% had B symptoms. RESULTS: Patients were treated with six to eight cycles of AMOPLACE and analyzed for response and survival. With a median follow-up of 48 months, complete responses (CRs) were seen in 68% of all patients with failure-free survival (FFS) and overall survival (OS) estimates at 4 years of 45% and 54%. In the DLCL subset, the CR rate was 69% and FFS and OS estimates at 4 years were 49% and 60%, respectively. The major toxicity was myelosuppression, with 73% of patients having WBC nadirs less than 1,000/microL; two treatment-related deaths occurred. CONCLUSION: We conclude that AMOPLACE is associated with CR and OS rates comparable with those of other third-generation regimens. PMID- 8426202 TI - Second malignancies after treatment of Hodgkin's disease: the influence of treatment, follow-up time, and age. AB - PURPOSE: In the period 1968 through 1988, The Norwegian Radium Hospital (NRH) treated an unselected population of 1,152 patients with Hodgkin's disease (HD) that comprised more older patients (mean age, 43 years) than most other institutions. We considered it important to evaluate these patients for development of second cancers (SCs). PATIENTS AND METHODS: The Norwegian Cancer Registry identified previously untreated patients with HD treated at NRH who had developed a SC more than 1 year after diagnosis of HD. The relative risk ratio (RR) (observed/expected cases) and the cumulative risk were calculated. RESULTS: Sixty-eight patients had developed a SC, including nine acute nonlymphocytic leukemias (ANLLs), eight non-Hodgkin's lymphomas (NHLs), and 51 solid tumors, including 11 lung cancers. The RR of SC and leukemia was 1.86 (95% confidence interval [CI], 1.4 to 2.4) and 24.3 (95% CI, 11.1 to 46.2), respectively. The RR of SC was highest in younger patients (< 41 years, RR = 3.8). No significant association between splenectomy and development of ANLL was found. The influence of treatment and follow-up time on the development of SC agrees with data from other large cancer institutions. CONCLUSION: (1) The low RR of developing a SC in this study is probably due to the number of older patients included, who have a lower RR of developing a SC due to less aggressive treatment, shorter follow-up time, and higher incidence of cancer in the expected background population. (2) The low RR and cumulative risk of developing ANLL may be due to the limited use of extensive chemotherapy (CT) in our hospital in the earlier years. PMID- 8426203 TI - Second malignant neoplasms in children treated for rhabdomyosarcoma. Intergroup Rhabdomyosarcoma Study Committee. AB - PURPOSE: This study was performed to determine the incidence and risk factors involved in the development of a second malignant neoplasm (SMN) after treatment of primary rhabdomyosarcoma (RMS) in patients enrolled onto Intergroup Rhabdomyosarcoma Studies I and II (IRS I and II). PATIENTS AND METHODS: There were 1,770 patients with primary RMS entered onto IRS I and II between 1972 and 1984. They were treated with chemotherapy and, in most instances, radiotherapy according to randomized or assigned regimens based on clinical grouping. Median follow-up time for these patients was 8.4 years. Incidence density (ID) was calculated for each study and for treatment and age groups. The 10-year cumulative incidence was estimated for each study. RESULTS: Twenty-two SMNs have been reported through 1991. The most common tumor type was a bone sarcoma followed by acute nonlymphoblastic leukemia (ANLL). The median time to the development of an SMN was 7 years (range, 1 11/12 to 15 9/12 years). The 10-year cumulative incidence rate was 1.7% for both studies. ID and cumulative incidence estimates were highest for patients who received both an alkylating agent and radiotherapy. The majority of patients for whom family histories were available had either neurofibromatosis themselves or a family history that suggested the Li Fraumeni syndrome (LFS). CONCLUSION: The results of this study suggest that genetic abnormalities play a prominent role in the development of an SMN after therapy for a primary RMS. Chemotherapy with an alkylating agent and radiotherapy play significant roles in the development of an SMN compared with patients who received only one of these therapeutic modalities. PMID- 8426204 TI - Treatment of CNS relapse in children with acute lymphoblastic leukemia: A Pediatric Oncology Group study. AB - PURPOSE: To assess the efficacy and toxicity of chemotherapy and cranial radiation for the treatment of children with acute lymphoblastic leukemia (ALL) following first isolated CNS relapse. PATIENTS AND METHODS: One hundred twenty children were treated on Pediatric Oncology Group (POG) protocol 8304. All children had received prophylactic CNS therapy during their initial treatment. The treatment protocol included a four-drug reinduction and six weekly doses of triple intrathecal therapy (TIT). Cranial radiation, 24 Gy, was followed by monthly TIT. Systemic consolidation and maintenance therapy included 6-week cycles of mercaptopurine/methotrexate (6MP/MTX) and vincristine/cyclophosphamide (VCR/CTX), with randomization to intervening pulses of prednisone/doxorubicin (PDN/DOX) or teniposide (VM26)/cytarabine (Ara-C) for a total of 88 weeks. RESULTS: All 120 patients achieved a second complete remission. There have been 61 protocol failures. Thirty-five patients had a bone marrow relapse, four with simultaneous CNS involvement and one with concurrent testicular leukemia. Thirteen patients had a second isolated CNS relapse, 10 a testicular relapse, and two relapsed in other sites. One patient died in remission. Overall event-free survival (EFS) at 4 years was 46% +/- 7%. The toxicity associated with this protocol was minimal except for leukoencephalopathy, which occurred in 20 (17%) patients. The treatment comparison between VM26/Ara-C or PDN/DOX pulses showed a trend toward superior EFS (P = .12) in favor of VM-26/Ara-C. CONCLUSION: To date, this represents the largest series of patients with ALL treated uniformly for an isolated CNS relapse. Since marrow relapse remains the primary site of failure, future protocols must intensify systemic therapy. PMID- 8426205 TI - Does cranial irradiation reduce the risk for bone marrow relapse in acute myelogenous leukemia? Unexpected results of the Childhood Acute Myelogenous Leukemia Study BFM-87. AB - PURPOSE: One of the goals of study AMA-BFM-87 was to test prospectively in acute myelogenous leukemia (AML) patients if cranial irradiation could be replaced by late intensification therapy with high-dose cytarabine (Ara-C) and etoposide (VP 16). PATIENTS AND METHODS: Patients with a low risk of CNS relapses (ie, no initial CNS disease, WBC count at diagnosis < or = 70.000/microL) were randomized for irradiation (group A, 31 patients). In 25 patients (group B), randomization was refused. As interim results showed no increase of CNS relapses in nonirradiated patients, prophylactic irradiation was discontinued after 2 1/2 years to prevent unnecessary CNS toxicity. Forty-four patients (group C) entered the study after randomization had been stopped. RESULTS: In all patients with a low risk of CNS recurrences (n = 100), a significantly higher probability of relapse-free interval (pRFI) of 5 years was found in irradiated patients (pRFI = .78) compared with nonirradiated patients (pRFI = .41) (P = .007). Moreover, a slightly higher incidence of CNS relapses was observed in nonirradiated patients. Due to the small number of patients, this was not observed when randomized patients only were analyzed. In accordance with these findings, the favorable outcome of low-risk patients in the preceding study, AML-BFM-83 (pRFI > .80), could only be reproduced in study AML-BFM-87 in patients who had received cranial irradiation. CONCLUSION: These results indicate that cranial irradiation should be an integral part of the treatment of all AML patients not undergoing bone marrow transplantation. Residual blasts in the CNS may escape systemic chemotherapy and lead to recurrence of the initial disease not only in the CNS, but also in the bone marrow. PMID- 8426206 TI - Escalating teniposide systemic exposure to increase dose intensity for pediatric cancer patients. AB - PURPOSE: The primary objective for this study was to determine whether controlling pharmacokinetic variability, by designing patient-specific dosage regimens for teniposide using a Bayesian estimation control strategy, would permit an increase in dose intensity without increased toxicity. PATIENTS AND METHODS: Twenty patients with relapsed acute lymphocytic leukemia were given teniposide as part of their induction and maintenance therapy. Before beginning reinduction therapy, an intensive pharmacokinetic study was performed based on 12 measured teniposide plasma concentrations. Doses were determined to achieve a targeted systemic exposure defined by an area under the plasma concentration time curve (AUC) beginning at an AUC consistent with that predicted for a patient with average pharmacokinetic parameters receiving the currently accepted maximal tolerated dose. The targeted systemic exposure was then escalated in increments of 25% in cohorts of at least three patients until unacceptable toxicity occurred. In 36 follow-up studies, when teniposide was administered during maintenance therapy, a Bayesian strategy based on only three or five measured drug concentrations was evaluated for precision and bias for achieving the targeted systemic exposure against the full pharmacokinetic study. RESULTS: Teniposide clearance varied over a fivefold range (3.7 to 21.6 mL/min/m2). With the use of the patient-specific dosage regimens, the intensity of systemic exposure was increased 50% (1,656 mumol.h v 1,060 mumol/L.h) over that previously possible with standard fixed doses, with no increase in acute, nonhematologic toxicity. Teniposide concentrations (n = 265) were well predicted (R2 = .82) with as few as three measured values from the initial study. CONCLUSION: Targeting systemic exposure is clinically feasible, precise, and allows increased dose intensity for teniposide without increased risk of acute, nonhematologic toxicity, when compared with fixed-dose regimens. PMID- 8426207 TI - Dose-escalation trial of M195 labeled with iodine 131 for cytoreduction and marrow ablation in relapsed or refractory myeloid leukemias. AB - PURPOSE: Mouse monoclonal antibody (mAb) M195 (anti-CD33) is reactive with most myeloid leukemia cells, monocytes, and hematopoietic progenitors, but not with other hematopoietic cells or stem cells nor with nonhematopoietic human tissues. A therapeutic dose-escalation study of M195 labeled with iodine 131 was conducted in patients with relapsed or refractory myeloid leukemias. METHODS: Twenty-four patients (16 relapsed or refractory acute myeloid leukemias, five blastic myelodysplastic syndromes [MDS], two chemotherapy-related secondary leukemias, and one blastic chronic myelogenous leukemia [CML]), including seven who had failed to respond to prior bone marrow transplantation (BMT), received from 50 mCi/m2 to 210 mCi/m2 of 131I-M195 in divided doses. RESULTS: In 22 patients, whole-body gamma-imaging demonstrated marked uptake of antibody into all areas of bone marrow. Twenty-three patients (96%) demonstrated decreases in peripheral blood cell counts, with decreased percentage of bone marrow blasts seen in 83% of cases. Eighty-nine percent of bone marrow biopsies examined quantitatively demonstrated substantial decreases in the number of blasts, with greater than 99% of blasts killed in some patients. The two cases that failed to demonstrate leukemic cytoreduction occurred in the first two dose levels. For 131I doses of 135 mCi/m2 or greater, pancytopenia was profound and lasted for at least 12 days. Eight patients had sufficient marrow cytoreduction to proceed to BMT. Three of these achieved marrow remission, one of 6+, and one of 9 months' duration. Two patients in blastic phase temporarily reverted to their original myelodysplastic states. Thirty-seven percent of assessable patients developed human anti-mouse antibody (HAMA). In two patients with HAMA who were re-treated, plasma 131I-M195 levels could not be maintained and no therapeutic effect resulted. Significant nonhematologic toxicity (hepatic) was seen in one patient and the maximum tolerated dose (MTD) was not reached. CONCLUSION: These data suggest that safe leukemic cytoreduction can be achieved with 131I-M195 even in multiply relapsed or chemotherapy-refractory leukemias. This agent may be useful as part of a preparative regimen for BMT. PMID- 8426208 TI - Second allogeneic marrow transplantation for patients with recurrent leukemia after initial transplant with total-body irradiation-containing regimens. AB - PURPOSE: The impact of a second marrow transplant on long-term disease-free survival (DFS) was evaluated for 77 consecutive patients aged 2 to 51 years who relapsed subsequent to allogeneic marrow transplantation after high-dose chemotherapy and total-body irradiation (TBI). PATIENTS AND METHODS: Patients received a second transplant for recurrent chronic myelogenous leukemia (CML) (n = 28), acute myelogenous leukemia (AML) (n = 32), and acute lymphoblastic leukemia (ALL) (n = 15) or lymphoma (n = 2) that used the same marrow donor as the initial transplant. High-dose chemotherapy of busulfan (BU) and cyclophosphamide (CY), or CY, carmustine (BCNU), and etoposide (VP-16), was used as a preparative regimen for the second transplant. Graft-versus-host disease (GVHD) prophylaxis consisted of the following: no prophylaxis (n = 8), T-cell depletion (n = 36), methotrexate (MTX) only (n = 21), cyclosporine (CSP) only (n = 1), MTX and CSP (n = 9), or anti-thymocyte globulin (ATG) and prednisone (n = 2). RESULTS: Engraftment occurred in the 74 assessable patients. Severe veno occlusive disease (VOD) was the most frequent cause of grades 3 and 4 regimen related toxicity (RRT); it occurred in 20 patients. The probability of death before day 100 from nonleukemic causes was 36%. The probability of relapse after second transplant was 70%, and the DFS rate was 14% (median DFS, 36 months; range, 22 to 87). The DFS rates for ALL, AML, and CML were 8%, 10%, and 25%, respectively. Multivariate analysis showed that the risk of relapse was inversely associated with acute GVHD (relative risk [RR] of relapse = 0.2; P = .0009). No other factor was associated with relapse. DFS was associated with the presence of acute GVHD (RR of treatment failure = 0.5; P = .0085), and a reduction of DFS was associated with severe VOD (RR = 10.6; P = .0001) and those patients older than 10 years (RR = 2.5; P = .0337). CONCLUSION: These data show that some patients may benefit from a second marrow transplant for recurrent leukemia after an initial marrow transplant. Younger patients and patients with CML especially should be considered as potential candidates for a second transplant. PMID- 8426209 TI - Autologous bone marrow transplant in acute myeloid leukemia in first remission. AB - PURPOSE: The Eastern Cooperative Oncology Group conducted a prospective study of postremission high-dose chemotherapy and autologous bone marrow transplantation (autoBMT) in a group of uniformly treated adults with de novo acute myeloid leukemia (AML) to evaluate whether intensive, myeloablative therapy in first complete remission (CR) could improve the disease-free survival. PATIENTS AND METHODS: After initial CR was induced by the combination of daunorubicin, cytarabine, and thioguanine, patients not eligible for allogeneic bone marrow transplantation (alloBMT) were offered autoBMT. Within a median of 2 months after CR, and without intervening postremission therapy, bone marrow was obtained, purged by exposure to 4-hydroperoxycyclophosphamide (4-HC), and cryopreserved. High-dose therapy consisted of oral busulfan over 4 days (16 mg/kg total) followed by intravenous (IV) cyclophosphamide 50 mg/kg daily for 4 days. The cryopreserved marrow was then reinfused. RESULTS: Of the 39 patients scheduled for autoBMT, four relapsed before transplantation. Two of the 35 (6%) transplant patients died of transplant-related complications, and 11 (33%) relapsed a median of 8 months after marrow reinfusion. No relapse has occurred after 24 months posttransplant. With a median follow-up of 31 months, the median disease-free survival period for all 39 patients has not been reached; however, 54% +/- 16% of patients are projected to be alive and disease-free at 3 years. CONCLUSION: Long term, disease-free survival after autoBMT in AML seems to be better than the outcome after conventional-dose postremission therapy and rivals the results of alloBMT. PMID- 8426210 TI - Harvesting bone marrow in an outpatient setting using newer anesthetic agents. AB - PURPOSE: To evaluate the results of outpatient bone marrow harvest (BMH). PATIENTS AND METHODS: Seventy-two adult patients with various malignancies had 79 BMH procedures performed for future autologous bone marrow transplantation (BMT) in our institution's outpatient surgical facility. All patients were evaluated and educated before the procedure. Newer anesthetic agents specifically developed to have shorter half-lives, more rapid recovery from general anesthesia, and fewer unpleasant side effects were chosen. Propofol was used for induction of anesthesia in 76 patients, the other three were induced with sodium pentothal. The blood volume removed was replaced by colloid (6% hydroxyethyl starch). Also, a new parenteral nonnarcotic pain medication, ketoroloc, was used during the last part of general anesthesia to help with expected postoperative pain in 76 patients. RESULTS: BMH took 111 +/- 24 minutes and patients were in postanesthesia care unit (PACU) for 220 +/- 72 minutes before being sent home with a companion and Tylenol with codeine (acetaminophen with codeine; McNeil Pharmaceutical, Spring House, PA). PACU complications were minor and included transient mild dizziness (7.6%), vomiting (3.8%), and fever (2.6%). No life threatening complication was observed. Only one patient was hospitalized for observation (fever) and then sent home. Seventy-five patients (94.9%) were contacted at home by the hospital nursing staff the day following the procedure. Five (6.7%) complained of nausea or vomiting, and four (5.3%) developed fever at home (temperature, 37.2 to 38.3 degrees C). Only 36% of patients actually took oral narcotic pain medication at home. CONCLUSION: Autologous BMH (AuBMH) is a safe outpatient procedure with minimal side effects when newer anesthetic agents are used. PMID- 8426211 TI - Randomized trial of hepatic arterial floxuridine, mitomycin, and carmustine versus floxuridine alone in previously treated patients with liver metastases from colorectal cancer. AB - PURPOSE: This study was designed to determine if hepatic arterial therapy with floxuridine (F), mitomycin, and carmustine (BCNU) (FMB) is superior to hepatic arterial therapy with F alone in previously treated patients with hepatic metastases from colorectal cancer. PATIENTS AND METHODS: Ninety-five patients were randomized to intrahepatic FMB versus intrahepatic F. All patients had tumor progression after systemic chemotherapy (either therapeutic or adjuvant). RESULTS: There was no significant difference in response rate (47% FMB v 33% F; P = .17). Median survival was similar in the two groups, 19.1 months for the FMB group compared with 14.0 months for the F group (P = .23). The overall median survival was 16.8 months. In patients who received prior adjuvant therapy, there was no difference between the two groups, but response rate was high in both (50% FMB v 62% F). The response rate for all patients who had received only prior adjuvant therapy versus all those who had received prior therapy for metastatic disease was 57% and 35%, respectively (P = .066). In the subset of patients whose disease had progressed with prior systemic chemotherapy, the response rate to FMB was greater than that to F (47% v 23%; P = .035). CONCLUSION: The overall partial response rate of 39% and the overall survival of 16.8 months from initiation of intrahepatitis therapy show that hepatic arterial therapy is a reasonable treatment option for patients whose tumor does not respond to systemic therapy or whose disease progresses after adjuvant therapy for colorectal cancer. PMID- 8426212 TI - Multicenter phase II efficacy trial of toremifene in tamoxifen-refractory patients with advanced breast cancer. AB - PURPOSE: To explore further the efficacy of high-dose toremifene in patients with advanced breast cancer who had failed to respond to tamoxifen or whose disease had progressed on tamoxifen. PATIENTS AND METHODS: One hundred two perimenopausal or postmenopausal women with metastatic breast cancer refractory to tamoxifen were entered onto a phase II clinical trial of toremifene at a dose of 200 mg/d. The study patients consisted of 28 primarily refractory patients; 43 patients who had relapsed after a prior tamoxifen response; and 31 patients who had relapsed while receiving adjuvant tamoxifen. This was a heavily pretreated group of patients, with 65% having failed chemotherapeutic attempts and 72% having failed two or more hormonal therapies. Forty-nine percent of patients had visceral dominant disease. RESULTS: The objective response rate was 5% (95% confidence interval [CI], 3% to 7%). The median time to treatment failure (TTF) was 10.9 months for the five responders. An additional 23% of patients had stable disease for a median TTF of 7.8 months, whereas the patients who experienced treatment failure had a median TTF of 2.1 months. Whether those patients with stable disease derived clinical benefit or simply had slow progression in an intrinsically indolent disease presentation is uncertain. Common toxicities were generally mild and similar to those encountered with tamoxifen. CONCLUSION: We conclude that there is major cross-resistance between tamoxifen and toremifene and that only occasional tamoxifen-refractory patients will have objective responses to toremifene. PMID- 8426213 TI - DNA ploidy and percent S-phase as prognostic factors in node-positive breast cancer: results from patients enrolled in two prospective randomized trials. AB - PURPOSE AND METHODS: To help clarify the clinical utility of flow-cytometric parameters, we performed flow cytometry on archival paraffin-embedded primary breast cancers from 502 patients treated on two adjuvant chemotherapy protocols performed by the North Central Cancer Treatment Group (NCCTG) and Mayo Clinic. DNA ploidy and percent S-phase (%S) were examined in univariate and Cox model multivariate analyses along with tumor size, menopausal and estrogen receptor status, Quetelet's index (QI), number of positive nodes and nodes examined, and Fisher and nuclear grades. RESULTS: Ploidy analysis showed that 40% of tumors were DNA diploid and 60% were DNA nondiploid (12% tetraploid and 48% aneuploid). There was no difference in relapse-free survival (RFS) (P = .82) or overall survival (OS) (P = .78) between the ploidy groups. Tetraploid patients had the longest RFS and OS of any group, but this did not achieve statistical significance. The %S was computed in 98% of cases and the medians were 9.0% for all patients, 6.4% for diploid patients, and 11.7% for nondiploid patients (P < .0001). By use of a %S greater than 12.3 as a prognostic variable in a univariate analysis, there was a significant difference in the RFS (P = .02) and OS (P = .007) of patients with low- versus high-proliferative tumors. However, when the %S was adjusted for clinical characteristics in the multivariate analysis, it was not a significant factor for RFS (P = .23) or OS (P = .36). CONCLUSION: These results indicate that DNA content and %S measurements by flow cytometry are not clinically useful independent prognostic factors in women with resected node positive breast cancer administered adjuvant chemotherapy. PMID- 8426215 TI - Multiple brain metastases are associated with poor survival in patients treated with surgery and radiotherapy. AB - PURPOSE: A retrospective analysis was performed to evaluate the role of surgery in the management of patients with solitary and multiple brain metastases. PATIENTS AND METHODS: Between 1980 and 1990, 46 patients underwent surgical resection of brain metastases at the University of Colorado Health Sciences Center. All but two patients received postoperative whole-brain radiotherapy to a median total dose of 30 Gy (range, 11.4 Gy to 50.0 Gy). Lung was the most common (56%) primary site and adenocarcinoma was the most common (46%) tumor histology. Twenty-eight of 46 patients (61%) had solitary metastases, while the remaining 18 patients had two or more foci. RESULTS: The median survival of all 46 patients was 11 months, and the 1- and 2-year survival rates were 40% and 12%, respectively. Moderately severe to severe neurologic impairment at the time of diagnosis and the presence of multiple brain metastases were associated with a significantly poorer survival. In patients with solitary metastasis, gross total resection and adenocarcinoma tumor histology significantly prolonged survival, whereas primary tumor site, the presence of active extracranial disease, and radiation dose had no significant effect on survival. CONCLUSION: These results are consistent with a recent randomized study supporting the role of surgery and whole-brain radiation therapy in the management of patients with solitary brain metastases. We would caution against the generalization of this concept to patients with two or more brain metastases. PMID- 8426214 TI - Cisplatin, fluorouracil, and leucovorin augmented by interferon alfa-2b in head and neck cancer: a clinical and pharmacologic analysis. AB - PURPOSE: To increase the activity of cisplatin, fluorouracil (5-FU), and leucovorin (PFL) through further biochemical modulation and study the pharmacologic interaction of 5-FU and interferon alfa-2b (IFN). PATIENTS AND METHODS: Escalating doses of IFN (0.5 to 4.0 x 10(6) U/m2/d x 6) were added to cisplatin 100 mg/m2, continuous infusion 5-FU 800 or 640 mg/m2/d x 5, and leucovorin 100 mg orally every 4 hours. Forty-eight previously untreated patients with locoregionally advanced head and neck cancer received up to three cycles of PFL-IFN. RESULTS: Twenty-one patients were treated during a phase I cohort study. Dose-limiting mucositis was seen with 800 mg/m2/d of 5-FU and 0.5 x 10(6) U/m2/d of IFN. After decreasing the 5-FU dose to 640 mg/m2/d, the maximally tolerated dose (MTD) of IFN was 2.0 x 10(6) U/m2/d. Mucositis and myelosuppression were dose-limiting. Of 34 patients treated at this MTD, 56% (95% confidence interval, 39% to 73%) had a complete remission. There was no correlation between 5-FU clearance and IFN dose. Pharmacodynamic analyses at the MTD showed that older age, female sex, and higher 5-FU area under the time versus concentration curve (AUC) were associated with lower nadir counts and/or increased mucositis. Seven patients with diabetes mellitus had significantly increased myelosuppression, serum creatinine, hypocalcemia, higher 5-FU concentrations, and lower 5-FU clearance compared with nondiabetics. CONCLUSION: The recommended doses for PFL IFN are 640 mg/m2/d for 5-FU and 2.0 x 10(6) U/m2/d for IFN. Sex, age, 5-FU AUC, and diabetes mellitus may have an impact on the pharmacodynamics of this regimen. PMID- 8426216 TI - Bioavailability of low-dose oral etoposide. AB - PURPOSE: To determine the bioavailability of oral etoposide capsules administered at doses of 100 mg and 400 mg. PATIENTS AND METHODS: The bioavailability of oral etoposide was determined by measuring the area under the etoposide plasma concentration versus time curve (AUC) following intravenous (IV) etoposide administration and comparing that value to the AUC achieved following an oral dose administered 1 day later to the same patient. The bioavailability of a 100 mg oral dose of etoposide was measured on 16 occasions in 11 patients. The bioavailability of a 400-mg dose was determined on 12 occasions in six patients. RESULTS: The mean (+/- SD) bioavailability following a 100-mg dose of oral etoposide was 76% +/- 22%, which was significantly greater (P < .01) than the mean bioavailability of 48% +/- 18% following a 400-mg oral dose. The coefficient of variation in oral etoposide bioavailability was significant: 29% with a 100-mg oral dose and 37% with a 400-mg dose. CONCLUSION: Bioavailability of a 100-mg oral etoposide dose is greater than suggested in the package insert from Bristol Laboratories (Evansville, IN). Comparable oral etoposide doses are not uniformly twice that of an IV dose, as suggested by the package insert, but will depend on the final oral dose administered. Bioavailability is better at lower oral etoposide doses. This study confirms the wide interpatient and intrapatient variability in oral etoposide bioavailability. PMID- 8426217 TI - Meta-analyses, use and misuse. PMID- 8426218 TI - More is not always better: a case for low-dose leucovorin. PMID- 8426219 TI - A new complication of permanent indwelling central venous catheters using high dose fluorouracil and leucovorin. PMID- 8426220 TI - The function of dendritic spines: devices subserving biochemical rather than electrical compartmentalization. PMID- 8426221 TI - Coordinate expression of c-fos and jun B is induced in the rat striatum by cocaine. AB - In cells in culture, specific stimuli induce selective patterns of immediate early gene induction. In the present study, we tested for such selectivity of stimulated gene expression by monitoring the expression of fos/jun gene mRNAs in the striatum in rats treated in vivo with the indirect dopamine agonist cocaine. We found by Northern blot and in situ hybridization analysis that cocaine induces the coordinate expression of c-fos and jun B mRNAs in neurons of the rat's striatum. By contrast, another immediate-early gene of the leucine-zipper family, c-jun, was not induced in striatal neurons by cocaine at any time tested from 1 to 24 hr after treatment. With the same probe, we could detect the induction of c jun mRNA (as well as that of c-fos and jun B mRNAs) in the hippocampus following administration of pentylenetetrazol. The induction of expression of c-fos and jun B was rapid and transient, with peak expression occurring at approximately 1 hr after cocaine administration, and the induction of the two genes was in similar striatal sites. These results establish that differential patterns of expression of fos/jun genes occur in striatal neurons following exposure to cocaine, a potent psychomotor stimulant. We suggest that these tissue-specific patterns of gene expression may contribute to the response specificity of striatal neurons to stimulation by monoamines including dopamine. PMID- 8426222 TI - Gonadotropin-releasing hormone-containing neurons change size with reproductive state in female Haplochromis burtoni. AB - In the preoptico-hypothalamic area (POA) of teleost fish, neurons containing gonadotropin-releasing hormone (GnRH) regulate reproduction through direct projections to pituitary gonadotropes. Here we show that these GnRH-containing cells change size depending on the reproductive and maturational state in female Haplochromis burtoni. We selected animals prior to, during, and after the reproductive portion of their life history, in both brooding and spawning states. Immunocytochemical staining of GnRH-containing neurons in the POA revealed that these cells are up to twice as large in females that have never spawned or are in the act of spawning than they are in females that are carrying broods. Older, postreproductive females have the largest cell sizes. Previous work on male H. burtoni has shown that soma sizes of the homologous neurons change according to social status, with dominant fish having larger cells than subordinates. Since reproductively active females have no apparent social hierarchy and are all exposed to approximately the same external stimuli, the primary factor(s) controlling GnRH-immunoreactive (irGnRH) neuron size appears to be internal reproductive state. Thus, while irGnRH neurons are pleiomorphic in both males and females, cell size change is differently regulated in each. PMID- 8426223 TI - Three subtypes of alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors are expressed in chick retina. AB - A recent report described the isolation of cDNA clones encoding alpha 7 and alpha 8 subunits of alpha-bungarotoxin-sensitive nicotinic ACh receptors (alpha BgtAChRs) from chick brain and demonstrated that they were related to, but distinct from, the alpha subunits of nicotinic ACh receptors (nAChRs) from muscles and neurons. Monoclonal antibodies against the two alpha BgtAChR subunits were used to demonstrate that at least two subtypes are present in embryonic day 18 chicken brain. The predominant brain subtype contains alpha 7 subunits, while a minor subtype contains both alpha 7 and alpha 8 subunits. Both subtypes may also contain other subunits. Here we report the results of immune precipitation studies and immunohistochemical studies of alpha BgtAChRs in the chick retina. In addition to the two subtypes found in brain, a new alpha BgtAChR subtype that contains alpha 8 subunits, but not alpha 7 subunits, was identified and was found to be the major subtype in chick retina. This subtype has a lower affinity for alpha-bungarotoxin (alpha Bgt) than does the subtype containing only alpha 7 subunits. Small amounts of this alpha 8 subtype were also detected in brain by labeling with higher concentrations of 125I-alpha Bgt than had been used previously. The subtype containing only alpha 7 subunits comprised 14% of the alpha BgtAChRs in hatchling chick retina. The subtype containing alpha 8 subunits (but no alpha 7 subunits) accounted for 69%, and the alpha 7 alpha 8 subtype accounted for 17%. Amacrine, bipolar, and ganglion cells displayed alpha 8 subunit immunoreactivity, and a complex pattern of labeling was evident in both the inner and outer plexiform layers. In contrast, only amacrine and ganglion cells exhibited alpha 7 subunit immunoreactivity, and the pattern of alpha 7 subunit labeling in the inner plexiform layer differed from that of alpha 8 subunit labeling. These disparities suggest that the alpha BgtAChR subunits are differentially expressed by different populations of retinal neurons. In addition, the distribution of alpha BgtAChR subunit immunoreactivity was found to differ from that of alpha-Bgt-insensitive nAChR subunits. PMID- 8426224 TI - Convergent force fields organized in the frog's spinal cord. AB - Microstimulation of the gray matter of the frog's spinal cord was used to elicit motor responses. Force responses were recorded with the frog's ankle clamped while EMG activity was monitored. The collections of force patterns elicited at different leg configurations were summarized as force fields. These force fields showed convergence to an equilibrium point. The equilibrium paths were calculated from the force fields with the leg clamped. These paths predicted free limb motion in 75% of trials. The force fields were separated into active and prestimulation resting responses. The active force field responses had a fixed position equilibrium. These active force fields were modulated in amplitude over time, although the balance and orientations of forces in the pattern remained fixed. The active fields grouped into a few classes. These included both convergent and parallel fields. The convergent force fields (CFFS) could be observed in deafferented preparations. Motoneuron (MN) activity underlying the force fields was marked using sulforhodamine. The marked activity covered several segments. Several simulations and MN stimulations show that topography, limb geometry, and random activation could not account for the results. It is likely that propriospinal interneurons distribute the activity that underlies the responses observed here. Experiments showed that CFFs that resemble those elicited by microstimulation also underlie natural behaviors. The full variety of fields revealed by microstimulation was larger than the repertoire elicited by cutaneous stimulation. It was concluded that fixed-pattern force fields elicited in the spinal cord may be viewed as movement primitives. These force fields could form building blocks for more complex behaviors. PMID- 8426225 TI - Axonal regeneration contributes to repair of injured brainstem-spinal neurons in embryonic chick. AB - Recent results have demonstrated complete anatomical and functional repair of descending brainstem-spinal projections in chicken embryos that underwent thoracic spinal cord transection prior to embryonic day 13 (E13) of the 21 d developmental period. To determine to what extent axonal regeneration was contributing to this repair process, we conducted experiments using a double retrograde tract-tracing protocol. On E8-E13, the upper lumbar spinal cord was injected with the first fluorescent tracing dye to label those brainstem-spinal neurons projecting to the lumbar cord at that time. One to two days later (on E10 E15), the upper to mid-thoracic spinal cord was completely transected. After an additional 7-8 d, a different second fluorescent tracing dye was injected into the lumbar cord at least 5 mm caudal to the site of transection. Finally, 2 d later on E19 to postnatal day 4, the CNS was fixed and sectioned. Brainstem and spinal cord tissue sections were then viewed with epifluorescence microscopy. In comparison to nontrasected control animals, our findings indicated that there were relatively normal numbers of double-labeled brainstem-spinal neurons after a transection prior to E13, whereas the number of double-labeled and second-labeled brainstem-spinal neurons decreases after an E13-E15 transection. In addition, at each subsequent stage of development from E10 to E12, there was a greater number of double-labeled brainstem-spinal neurons (indicating regeneration of previously severed axons) than cell bodies labeled with the second fluorescent tracer alone (indicating subsequent development of late brainstem-spinal projections). Assessment of voluntary open-field locomotion (hatchling chicks) and/or brainstem evoked locomotion (embryonic or hatchling) indicated that functional recovery of animals transected prior to E13 was indistinguishable from that observed in control chicks (sham operated or unoperated). Taken together, these data suggest that regeneration of previously axotomized fibers contributes to the observed anatomical and functional recovery after an embryonic spinal cord transection. PMID- 8426226 TI - Functional studies of Alzheimer's disease tau protein. AB - In vitro assays were used to monitor and compare the kinetic behavior of bovine tubulin polymerization enhanced by tau proteins isolated from Alzheimer's disease (AD) and nondemented (ND) age-matched control brains. Tau from AD cases induced slower polymerization and a steady state turbidity value approximately 50% of that stimulated by tau from control cases. Tau from the most severe AD case was least effective at promoting polymerization. Dark-field light microscopy of the control samples revealed abundant microtubule formation and many microtubule bundles. Microtubule assembly was observed in AD samples as well, but bundling was not obvious. These results were confirmed by negative-stain electron microscopy. Morphological analysis showed that AD tau-induced microtubules were longer than control microtubules. Furthermore, our initial results suggest that the reduction of AD tau activity is correlated with neurofibrillary pathology in AD brains. Earlier reports indicated that AD tau is modified by phosphorylation (Grundke-Iqbal et al., 1986; Wood et al., 1986; Iqbal et al., 1989; Brion et al., 1991a,b; Lee et al., 1991). Our results support the hypothesis that tau modification compromises its function by altering its ability to nucleate and bundle microtubules. PMID- 8426227 TI - Dystrophin in a membrane skeletal network: localization and comparison to other proteins. AB - We studied the location, relative abundance, and stability of dystrophin in clusters of ACh receptors (AChRs) isolated from primary cultures of neonatal rat myotubes. Although variable amounts of dystrophin were found at receptor clusters, dystrophin was always associated with organized, receptor-rich domains (AChR domains). Dystrophin was occasionally seen in focal contact domains, but never in clathrin-coated domains. Dystrophin was also present in a diffuse, punctate distribution in regions of myotube membrane that did not contain AChR clusters. Immunogold labeling at the ultrastructural level localized dystrophin in a spectrin-rich filamentous network closely applied to the cytoplasmic surface of the cell membrane at AChR domains. Dystrophin was not associated with overlying actin filaments. Semiquantitative immunofluorescence studies indicated that dystrophin was present in relatively small amounts in these preparations, with only one molecule of dystrophin for every approximately 5 AChR, 43 kDa and 58 kDa molecules, and for every approximately 20-35 beta-spectrin molecules. Clusters were disrupted, but the total amount of dystrophin was not significantly reduced, when myotubes were incubated with sodium azide or in Ca(2+)-free medium, and when isolated AChR clusters were extracted at low ionic strength, at high pH, or in 6 M urea. These treatments extract other peripheral membrane proteins from AChR clusters. Labeling for dystrophin was completely eliminated when clusters were incubated with chymotrypsin, however. Thus, dystrophin forms part of a membrane skeleton at AChR clusters, but it is more difficult to remove than other proteins in the network. This suggests that dystrophin attaches to cluster membrane in a unique way. PMID- 8426228 TI - NGF induces neonatal rat sensory neurons to extend dendrites in culture after removal of satellite cells. AB - Vertebrate sensory neurons have a pseudo-unipolar morphology; their somata are covered by satellite cells and lack dendrites or synaptic contacts. However, when neonatal rat sensory neurons from the nodose ganglia develop in culture in absence of satellite cells and with NGF, they form synapses among themselves. In this study, we investigated whether neonatal rat nodose neurons express dendrites under the same culture conditions. We show by Lucifer yellow injection that nodose neurons remain typically unipolar when cocultured with their ganglionic satellite cells. However, when these neurons are cultured without satellite cells, virtually all neurons acquire a multipolar morphology. Moreover, when NGF is added to satellite cell-free cultures, several neurons extend dendrites; these processes stain positively for microtubule-associated protein-2. NGF induces a 17 fold increase in dendritic outgrowth after 3 weeks but has little effect on axon number. In addition, we find that the ability of nodose neurons to extend dendrites is developmentally regulated. Furthermore, in a combined morphological and electrophysiological study, using whole-cell voltage-clamp technique with Lucifer yellow in the recording solution, we demonstrate a positive correlation between the extent of dendritic outgrowth and the density of ACh currents, suggesting that these dendrites have ACh receptors. Our results indicate that neonatal rat nodose neurons are capable of extending dendrites and that extrinsic factors can induce or suppress their extension. In addition, the results suggest that these dendrites may act as principal post-synaptic structures for synapse formation that occurs in these cultures. PMID- 8426229 TI - Recovery of frontal cortex-mediated visual behaviors following neurotrophic rescue of axotomized neurons in medial frontal cortex. AB - Unilateral lesions extending across the boundary region of visual and parietal cortex in adult rats result in the death of 20-35% of neurons in layers II-III of the caudal third of medial frontal cortex ipsilaterally, a neuron population labeled with 3H-thymidine on the 19th day of gestation (E19). Additionally, there is a consistent 15% loss of these labeled neurons in an area between 50% and 60% of the distance along the caudal-rostral extent of medial frontal cortex, an area that may function analogously to the frontal eye field of primates. All of these neurons are rescued from axotomy-induced death by delivering into the posterior cortex lesion cavity for 2 weeks a macromolecular fraction of culture medium conditioned by embryonic primordia of the frontal-occipital pathway (CM). Moreover, the rescue is apparently permanent, with normal numbers of these neurons present in CM animals 6-7 weeks after the neurotrophic factor is no longer being supplied exogenously. Behaviorally, control operates receiving a similarly prepared fraction of unconditioned medium are significantly impaired in the number of trials needed to learn two visual discrimination tasks. This deficit is attributable in part to a bias in erroneous responses to the side contralateral to the lesion. The error bias reflects a failure to inhibit repeated incorrect responding contralaterally. In contrast, the CM animals learn both visual tasks in a normal number of trials and have no contralateral error bias. Rather, all CM animals have an contralateral error bias. Rather, all CM animals have an ipsilateral error bias (interpreted as an unmasking of the contralateral neglect expected after a parietal cortex lesion).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426230 TI - Confocal microscopy reveals coordinated calcium fluctuations and oscillations in synaptic boutons. AB - Calcium ions are one of the main factors regulating quantal transmitter release and thus synaptic transmission in the nervous system. Using confocal microscopy, fluorescent imaging with the calcium indicator Rhod-2, and time series analysis, we show that the levels of calcium ions inside single synaptic boutons of the lizard neuromuscular junction are not constant at rest, but undergo coordinated fluctuations in the space domain, which cover a large fraction of the synaptic bouton. Furthermore, oscillations in intracellular calcium were frequently observed in the time domain. Control experiments showed no coordinated fluctuations or oscillations at locations outside the synaptic boutons. Edge detection analysis showed that the coordinated fluctuations and oscillations were not due to movement artifacts. No coordinated fluctuations and oscillations were seen when similar measurements and analyses were performed on artificial fluorescent beads. A variance analysis was performed on artificial fluorescent beads and on synaptic boutons. The variance of the fluorescent signal at the synaptic boutons was larger than the variance in artificial beads with the same mean fluorescence. This extra variance was greatly reduced when the extracellular calcium concentration was decreased from 2.0 mM to 0.4 mM. We conclude that the coordinated fluctuations and oscillations in the calcium-induced fluorescence at the synaptic boutons are genuine biological phenomena and may be of significance in the regulation of transmitter release. PMID- 8426231 TI - Evidence for presynaptic inhibition of the olfactory commissural pathway by cholinergic agonists and stimulation of the nucleus of the diagonal band. AB - We have investigated the role of the projection from the magnocellular basal forebrain to the olfactory bulb in regulating synaptic transmission in the commissural connection between the two olfactory bulbs. Commissural fibers arise in the contralateral anterior olfactory nucleus, travel in the anterior wing of the anterior commissure (AC), and terminate in the granule cell layer of the olfactory bulb. Electrical stimulation of the commissure causes synaptic activation of granule cells in the granule cell layer of the bulb; the resulting field potential is a reliable indicator of this synaptic current. Microinjections of cholinergic agonists, but not of identical, or larger, quantities of vehicle, reduced the amplitude of this AC field potential. Systemic injection of scopolamine reversed this depression and returned the AC response amplitude to control levels. Irreversible AChE inhibition also reduced the amplitude of the AC response, and muscarinic blockade reversed this effect. Cholinergic terminals in the olfactory bulb arise entirely from the axons of magnocellular basal forebrain neurons in the nucleus of the diagonal band (NDB). Electrical stimulation of NDB, which should release ACh, as well as other transmitters, depressed the AC response. Brief trains of NDB shocks caused a moderate decrease in the AC response that lasted 1-2 sec. Longer shock trains, which caused marked potentiation of the NDB field potential, caused a profound, prolonged (> 20 sec) inhibition of the AC response. Antidromic tests demonstrated that NDB stimulation significantly decreased the excitability of AC terminals. This and other characteristics of the inhibition strongly suggest that the decrease in amplitude of the field potential response to AC stimulation caused by cholinergic agonists and stimulation of NDB is due to presynaptic inhibition leading to reduced release of transmitter from AC terminals. These results suggest that one function of the basal forebrain projection to the olfactory bulb is inhibition of the commissural connection between the two olfactory bulbs. As NDB has been implicated in theta pacemaker input to the olfactory bulb, phasic NDB inhibition of centrifugal afferents to the bulb could function to coordinate signal processing temporally in the olfactory system. Temporal coordination may be particularly important to olfactory circuit function, as this system lacks the point-to-point topographical organization characteristic of other sensory systems. PMID- 8426232 TI - Hypothalamic lesions that induce female precocious puberty activate glial expression of the epidermal growth factor receptor gene: differential regulation of alternatively spliced transcripts. AB - Injury of the nervous system triggers a complex series of repair mechanisms that include production of neurotrophic and mitogenic factors by cells neighboring the injured area. While trauma of most parts of the brain results in loss of function, lesions of certain regions of the female hypothalamus enhance the secretory activity of a group of specialized neurons that produce luteinizing hormone-releasing hormone (LHRH), the neuropeptide that controls sexual development. The increased output of LHRH causes sexual precocity by prematurely activating the neuroendocrine reproductive axis. Recent studies have implicated transforming growth factor alpha (TGF alpha) produced by reactive astrocytes in the process by which lesions hasten sexual maturation, and have suggested that the stimulatory actions of TGF alpha on LHRH neurons require the intermediacy of epidermal growth factor receptors (EGFRs). In the present study, we examined the changes in EGFR gene expression following lesions of the preoptic-anterior hypothalamic area (POA-AHA) of immature female rats, identified the cell types where EGFR synthesis increases, and assessed the biochemical activity of the newly formed EGFR protein. RNase protection assays demonstrated that the lesion significantly increased the levels of a predominant mRNA transcript encoding the full-length, membrane-spanning EGFR, but did not affect those of a much less abundant, alternatively spliced mRNA that encodes a truncated, presumably secreted form of EGFR. Following lesions, antibody-induced EGFR kinase activity increased twofold. Antibodies directed against a peptide sequence contained within the carboxy terminus of EGFR showed intense EGFR immunoreactivity in cells surrounding the lesion site; double immunohistochemistry identified these cells as astrocytes since EGFR immunoreactivity was colocalized with that of glial fibrillary acidic protein, an astrocytic marker. That these changes result from an increase in EGFR gene expression was indicated by the elevated levels of EGFR mRNA detected by in situ hybridization in cells of the same area. Although POA AHA lesions did not result in appearance of EGFR in LHRH neurons themselves, EGFR positive cells and processes were seen in close proximity to LHRH neurons and their nerve terminals, particularly in the area surrounding the lesion. Since TGF alpha gene expression is also increased in reactive astrocytes of POA-AHA lesions and blockade of EGFR prevented the advancing effect of the lesion on puberty (Junier et al., 1991b), the present results support the concept that, in lesioned animals, TGF alpha stimulates LHRH secretion indirectly via a paracrine mechanism that involves its interaction with EGFRs located on astroglial cells. PMID- 8426233 TI - Growth cone choices of Drosophila motoneurons in response to muscle fiber mismatch. AB - In Drosophila embryos, each motoneuron is accurately matched to one or more singly identifiable muscle fibers. In this article we altered the number and pattern of the embryonic muscle fibers using genetic, heat shock, and laser ablation methods to test whether motoneuron growth cones are able to recognize specific targets. The choices made by two motoneurons were assayed using both intracellular dye fills and immunocytochemistry. The motoneurons RP1 and RP3 have nearly identical central and peripheral axonal trajectories. However, RP3 innervates the two most ventral longitudinal muscle fibers, 7 and 6, while RP1 grows past these fibers to innervate only muscle fiber 13. In rhomboid mutants muscle fiber 7 does not develop. Despite the loss of one of its targets, RP3 faithfully innervated the remaining muscle fiber 6 in over 80% of the observed cases. Furthermore, neuron RP1 accurately innervated muscle fiber 13, although it traversed one fiber fewer to reach it. Laser ablation of muscle fiber 7 confirmed the target choices shown by the motoneurons. In numb mutants, multiple muscle fibers, including 7, 13, and 12, fail to develop. This allowed us to test whether fibers distal to the target are involved in muscle fiber recognition, possibly by halting the growth cone advance. In mutant embryos, RP3 innervated muscle fiber 6 at the same frequency regardless of the absence of the distal muscle fiber 13. By contrast, RP1, which had lost its target entirely, frequently failed to innervate any muscle fiber during the period examined. Finally, muscle fiber 13 can be duplicated in wild-type embryos by means of a brief heat pulse during myogenesis. Presented with two targets, RP1 innervated both fibers in each case examined, while RP3 synapsed with muscle fibers 7 and 6 normally. Neuron-specific antibodies revealed that the embryonic growth cone choices were not transient, but persisted into the larval neuromuscular projections. These results indicate that each motoneuron growth cone has a primary target preference, which is retained even when the numbers of the muscle fibers, and therefore their relative positions, are altered. We therefore suggest that synaptic recognition by Drosophila motoneuron growth cones relies on unique features of the individual muscle fibers. PMID- 8426234 TI - Characterization and topography of high-affinity 125I-neurotrophin-3 binding to mammalian brain. AB - The binding of biologically active and 125I-labeled neurotrophin-3 (NT-3) was studied with both dry film and emulsion autoradiography to compare with NGF binding and discover areas where NT-3 may function in vivo. The equilibrium binding of 300 pM 125I-NT-3 to rat brain sections was reversible and inhibited by unlabeled NT-3 (IC50, 420 pM). 125I-NT-3 bound in a saturable manner, with high affinity (Kd, 227-269 pM), and with a capacity (Bmax, 26 fmol/mg protein) that exceeded that of NGF by threefold. As with NGF, 125I-NT-3 also bound to a second population of sites with lower affinity (Kd, 2.8 nM) and higher capacity (Bmax, 170 fmol/mg protein). 125I-NT-3 binding was not blocked by NGF, or serum proteins, and brain-derived neurotrophic factor (BDNF) competed for it in a distinctly biphasic manner (IC50 values of 230 pM and 37 nM). Microdensitometry confirmed graphically and by Hill analysis the monophasic displacement of 125I-NT 3 and the biphasic displacement of 125I-NT-3 binding by BDNF in hippocampus, caudate-putamen, neocortex, and olfactory tubercle. In rat or cat, the topography of 125I-NT-3 binding differed from that reported for 125I-NGF binding or for the low-affinity NGF receptor. The highest binding densities were found in neocortical layers 1 and 2, the stratum oriens and radiatum of hippocampus, molecular layer of the dentate gyrus, nucleus of the lateral olfactory tract, entorhinal cortex, anterior olfactory nucleus, anteromedial thalamic nucleus, and amygdala. Moderate densities were found in neocortical layers 4-6, the neostriatum, amygdala, the dorsal root ganglia, and the central gray of spinal cord. Emulsion autoradiography also revealed binding in nerve terminal-rich regions of superficial neocortex and hippocampus but not on neural cell bodies. Binding was absent in many other brain regions, including cholinergic nuclei, and in all peripheral organs studied including liver, kidney, pancreas, heart, and skeletal muscle. 125I-NT-3 binding to sections of human basal ganglia resembled that seen in rat or cat, including high densities in the caudate, putamen, and superficial neocortex. The unique distribution and pharmacology of 125I-NT-3 binding to BDNF-sensitive and -insensitive sites in brain predict predominantly neuronal actions for these factors that are likely to be more widespread and distinct from those of NGF. PMID- 8426235 TI - Independent guidance of retinal axons in the developing visual system of Drosophila. AB - The development of the adult visual system of Drosophila requires the establishment of precise retinotopic connections between retinal photoreceptor cell axons and their synaptic partners in the optic lobe of the brain. To assess the role of axon-axon interactions in retinal axon guidance, we used genetic methods to disrupt the normal spatiotemporal order of retinal axon ingrowth. We examined retinal axon projections to the developing first optic ganglion, the lamina, in two mutants in which reduced numbers of ommatidia develop in the eye imaginal disk. We find that in the developing lamina of these mutants, sine oculis and Ellipse, retinal axons project to proper dorsoventral positions despite the absence of the usual array of neighboring retinal axons. In a second approach, we examined animals that were somatic mosaics for the mutation, glass. In glass- animals, retinal axons project aberrantly and the larval optic nerve is absent. We find that in the developing lamina of glass mosaic animals, wild-type retinal axons project to proper dorsoventral positions despite the misrouted projections of neighboring glass- retinal axons. In addition, wild-type retinal axons project normally in the absence of the larval optic nerve, indicating that the latter is not an essential pioneer for retinal axon navigation. Our observations support the proposal that axon fascicles can make at least some pathfinding decisions independently of other retinal axon fascicles. We suggest that positional guidance cues that might label axon pathways and target destinations contribute to retinotopic pattern formation in the Drosophila visual system. PMID- 8426236 TI - Photic regulation of peptides located in the ventrolateral subdivision of the suprachiasmatic nucleus of the rat: daily variations of vasoactive intestinal polypeptide, gastrin-releasing peptide, and neuropeptide Y. AB - We have determined, by enzyme immunoassay, daily and circadian patterns of the concentrations of three peptides, which are located in the ventrolateral subdivision of the suprachiasmatic nucleus (SCN): vasoactive intestinal polypeptide (VIP), gastrin-releasing peptide (GRP), and neuropeptide Y (NPY). The contents of VIP and GRP, which are synthesized in the SCN, did not show circadian rhythms in constant darkness (DD). Under light-dark (LD) conditions, GRP content increased and VIP content decreased over the course of the light period and then gradually recovered during the dark period. Responsiveness of these peptides to light suggests that VIP and GRP may transmit visual information on duration of illumination. NPY, which is transported from the intergeniculate leaflet of the lateral geniculate body, showed a circadian rhythm with a peak at circadian time 12 hr in DD. This endogenous rhythm was remarkably modulated by photic stimulation. Under LD conditions, the NPY content in the SCN exhibited a bimodal rhythm with peaks at both the light-dark and dark-light transition points. Thus, NPY may convey visual information on the transitions. All these results indicate that the levels of VIP, GRP, and NPY are mainly regulated by light stimulation and suggest that peptides in the ventrolateral SCN are involved in the mediation of photic information to the pacemaker. PMID- 8426237 TI - Neurogenesis and differentiation of sympathetic B and C cells in the bullfrog tadpole. AB - The relation between birthdates of sympathetic neurons and their subsequent differentiation into cutaneous B cells and vasomotor C cells was examined in paravertebral ganglia 9 and 10 of the bullfrog tadpole. Neurons undergoing terminal cell division were identified by injecting tadpoles repeatedly with 5' bromodeoxyuridine (BRDU) for one to six developmental stages between III and XXI. After allowing the tadpoles to enter late metamorphic stages (XX-XXV), the ganglia were double immunostained for BRDU and neuropeptide Y (NPY). NPY is a marker for mature C-type neurons (Horn et al., 1987). Double-labeled neurons were readily discerned through use of distinct black and brown HRP reaction products and also because immunoreactivity for BRDU was localized in nuclei while that for NPY was localized in perinuclear cytoplasm. Counts of labeled cells showed that neurogenesis occurs throughout limb bud and paddle stages, and that it ceases during early foot stages (XII-XIV), a time coinciding with the onset of NPY expression. By contrast, the labeling of non-neuronal satellite cells with BRDU was most common weeks later during metamorphic stages. Irrespective of their birthdates, about half of the BRDU-labeled neurons were also positive for NPY immunoreactivity. This proportion of NPY-positive cells is indistinguishable from that in the entire adult ganglia (Horn et al., 1987). In addition to establishing that neurogenesis and gliogenesis occurs during tadpole stages, the results indicate that the onset of NPY expression by vasomotor C neurons is unrelated to their time of origin. In other words, the last wave of neurogenesis in sympathetic ganglia does not give rise to a specific subclass of sympathetic neurons. PMID- 8426238 TI - Synaptic potentials in the central terminals of locust proprioceptive afferents generated by other afferents from the same sense organ. AB - Afferent neurons from a proprioceptor [the femoral chordotonal organ (FCO)] at the femoro-tibial joint of a locust hindleg carry patterns of spikes to the CNS in which information is coded about the positions and movements of the tibia. Intracellular recordings from the afferents of this organ as they enter the CNS reveal spikes and depolarizing post-synaptic potentials (PSPs) during voluntary or imposed movements of the joint. Some of these PSPs are generated as a result of spikes in other FCO afferents, and can be evoked experimentally by electrical stimulation of the nerve from the organ. One afferent does not appear to synapse directly on another, but instead activates reliable pathways involving other central neurons. Current clamping of individual afferents in isolated ganglia shows that the PSPs are increased in amplitude by hyperpolarizing currents injected into an afferent, and decreased by depolarizing ones. They reverse at about -68 mV (n = 5). At the normal resting potential of the afferents, -72 mV (+/- 0.42 SE, n = 57), the PSPs are therefore depolarizing, and are associated with an increased conductance of the membrane. The changes in membrane potential and conductances associated with the PSPs can be mimicked by pressure injection of GABA into the regions of neuropil that contain the terminals of the afferents. The potential evoked by GABA is associated with an increased conductance of the membrane and reverses at the same potential as the PSPs. GABA also reduces the PSPs evoked in the terminals, either by movements of the FCO or by electrical stimulation of its nerve. The PSPs and the effects of the GABA-evoked potentials are mimicked by the GABA agonist muscimol. The PSPs are blocked reversibly by picrotoxin. The PSPs and the GABA-evoked potentials both alter the excitability of an afferent terminal by reducing the ability of the membrane to support an action potential. It is suggested that the PSPs are depolarizing, inhibitory potentials generated in the terminals of the afferents by central neurons that release GABA, and that their role is to change the efficacy of the afferent spikes at their first output synapses in the CNS. These interactions could form a graded, gain control mechanism for synaptic transmission at the afferent output synapses that is directly dependent on the features of the mechanical movements of the joint. PMID- 8426239 TI - Cell cycle parameters and patterns of nuclear movement in the neocortical proliferative zone of the fetal mouse. AB - Cytogenesis is the critical determinant of the total number of neurons that contribute to the formation of the cerebral cortex and the rate at which the cells are produced. Two distinct cell populations constitute the proliferative population, a pseudostratified ventricular epithelium (PVE) lying within the ventricular zone (VZ) at the margin of the ventricle, and a secondary proliferative population that is intermixed with the PVE within the VZ but also is distributed through the overlying subventricular and intermediate zones of the cerebral wall. The present analysis, based upon cumulative S-phase labeling of the proliferative cells with 5-bromo-2'-deoxyuridine, is principally concerned with the PVE of the gestational-day-14 (E14) murine cerebral wall. It has immediate but also more far reaching general objectives. The most immediate objective, essential to the design and interpretation of later experiments, is to provide estimates of critical parameters of cytogenesis for the PVE. The growth fraction is virtually 100%. The lengths of the overall cell cycle, S-, G2+M-m, and G1-phases are 15.1 hr, 3.8 hr, 2 hr, and 9.3 hr, respectively. The PVE is homogeneous with respect to cell cycle length. For methodological considerations, these estimates are more accurate than estimates of the same parameters obtained in earlier analyses based upon S-phase labeling with tritiated thymidine. It is particularly with respect to a shorter length of S-phase determined here that the present values are different from those obtained with thymidine. At a more innovative level, the temporal and spatial resolution of nuclear movement made possible by the methods developed here will allow, in a way not previously attempted, a fine-grained tracking of nuclear movement as cells execute the successive stages of the cell cycle or exit the cycle subsequent to mitosis. Such observations are pertinent to our understanding of the regulatory mechanisms of neocortical histogenesis and the cell biological mechanisms that govern the proliferative cycle of the ventricular epithelium itself. It is known that the velocity of nuclear movement in the PVE is maximum in G2 (fourfold increase from S-phase) and minimum in M and early G1.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8426240 TI - In vivo observations of pre- and postsynaptic changes during the transition from multiple to single innervation at developing neuromuscular junctions. AB - Synaptic rearrangements in developing muscle were studied by visualizing individual neuromuscular junctions in the sternomastoid muscle of living neonatal mice as they underwent the transition from multiple to single innervation. Vital staining of ACh receptors (AChRs) with rhodamine-conjugated alpha-bungarotoxin showed that while junctions were still multiply innervated (usually by two motor axons), regions of the postsynaptic membrane within each junction became depleted of receptors. Usually, several small postsynaptic areas lost AChRs in succession. In these areas, AChRs already in the membrane rapidly disappeared compared to a low level of receptor turnover elsewhere in the junction. Moreover, there was no evidence of new AChRs being inserted into these areas. Within each postsynaptic area undergoing AChR depletion, the intensity of receptor staining decreased gradually over 1-2 d. In some junctions, it appeared that AChRs were migrating away from areas being depleted of receptors. The depletion of AChRs from some sites in combination with the spreading apart of the entire receptor-rich area due to muscle fiber growth accounts for the transformation from plaque-like to branched receptor distributions at developing neuromuscular junctions. Vital staining of presynaptic motor nerve terminals at junctions whose postsynaptic AChRs were also stained showed that motor nerve terminals were lost from the same areas that were depleted of receptors postsynaptically. Postsynaptic areas began to be depleted of AChRs before there was any obvious loss of membrane or intracellular staining in the overlying nerve terminal. Only when a single innervating axon remained at a junction did loss of motor nerve terminals and underlying AChRs largely cease. That former synaptic areas could at later times be identified as uninnervated regions within a junction indicates that synapse elimination during development leaves an indelible mark on synaptic structure. These observations suggest that the withdrawal of a motor axon from a neuromuscular junction occurs as a consequence of the stepwise elimination of all of its synapses with that muscle fiber. These results also suggest that an important aspect of synaptic competition leading to axon withdrawal is the precocious loss of AChRs beneath the nerve terminals of the axon that will be eliminated. A similar early loss of AChRs beneath one axon's synapses has been shown to occur during synapse elimination in reinnervated adult muscle (Rich and Lichtman, 1989a).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8426241 TI - Regional differences in microtubule dynamics in the axon. AB - We have used an indirect method to compare the dynamic properties of microtubules (MTs) in the main shaft and distal regions of the axon. Individual MTs are staggered along the length of the axon and consist of a labile domain situated at the plus end of a stable domain (Baas and Black, 1990). As a result of this organization and the plus-end-distal orientation of axonal MTs, the most distal region of the axon consists entirely of labile domains, while the main shaft consists of a mixture of labile and stable domains. In this study, we wished to determine whether the labile domains extending into the distal axon differ in their dynamic properties from the labile domains terminating in the main shaft. To address this issue, we used immunoelectron microscopy to compare the tyrosination state of the labile domains terminating in these 2 axon regions. Because detyrosination is a polymerspecific modification of alpha-tubulin that accumulates with time, the levels of tyrosinated alpha-tubulin will be a reflection of the age, and hence dynamic properties, of the polymer. To maximize our chances of visualizing potential differences, we varied the concentration of the primary antibody in these experiments. Our studies indicate that the stable domains are generally deficient in tyrosinated alpha-tubulin, while the labile domains contain clearly detectable levels. Within the labile domain, the subsection closer to the plus end of the MT contains relatively higher levels of tyrosinated alpha-tubulin than does the subsection farther from the plus end, suggesting that the levels of tyrosinated alpha-tubulin in the labile domain may gradually increase as one moves away from the stable domain toward the plus end of the MT. Although these observations apply to the labile domains in both regions of the axon, the labile domains extending into the distal region contain comparatively higher levels of tyrosinated alpha-tubulin than do the labile domains terminating in the main shaft. These results are consistent with the view that highly dynamic MT polymer is present throughout the axon, but that the polymer nearest the advancing growth cone is particularly dynamic. PMID- 8426242 TI - Occupational illness in Oklahoma. AB - This study estimates the extent of work-related chronic disease fatalities in Oklahoma. Occupational cancer, pneumoconiosis, and chronic respiratory, cardiovascular, renal, and neurological diseases are addressed specifically. Also, the costs of chronic occupational illness are estimated. Because many cases of work-related disease find their way to the primary care physician, an individual who often has little formal training in the recognition and diagnosis of occupational or environmental illness, the education of primary care physicians and medical students in occupational disease recognition and prevention is encouraged. PMID- 8426244 TI - Injury control and prevention. PMID- 8426243 TI - Leaders in medicine: Mark R. Johnson, MD. PMID- 8426245 TI - More on spider bites and stun guns. PMID- 8426246 TI - Symposium on antimicrobial therapy. IV. The cephalosporins. AB - Hopefully this review has brought some cephalosporin contentment to replace cephalosporin confusion. From the classification of these antibiotics in Table 1, we have made some significant reductions. One should know how to use cefazolin for staphylococcal/streptococcal infections and for surgical prophylaxis. One should know that cephalexin is massively overused, and really now not all that useful an agent. Cefuroxime is a useful agent for beta-lactamase producing H. influenzae infections. Cefotetan has a role in surgical prophylaxis in ob/gyn and represents the best antianaerobic activity of the cephalosporins; although no cephalosporin is a primary drug for anaerobic infections. Cefuroxime axetil or cefprozil can be useful for comparatively minor infections due to beta-lactamase producing H. influenzae. A third generation cephalosporin represents a reasonable alternative, in certain situations, to aminoglycoside therapy for infections due to multiply drug-resistant Gram-negative bacilli. Ceftazidime is an alternative antipseudomonal beta-lactam antibiotic. Despite the lack of indications for use of cephalosporins as drugs of choice, rational use of these agents can provide safe, effective, and efficient therapy for a variety of infectious diseases. They will likely remain an important part of the physicians' antimicrobial armamentarium for the foreseeable future. PMID- 8426247 TI - Looking out for the cocaine abuser. PMID- 8426248 TI - Effect of flurbiprofen on tissue levels of immunoreactive bradykinin and acute postoperative pain. AB - This study evaluates whether preoperative administration of flurbiprofen alters the levels of immunoreactive bradykinin (iBK) peripherally released into inflamed tissue. Thirty-six patients were randomly treated on a double-blind basis with either flurbiprofen (100 mg) or placebo before the surgical extraction of impacted third molars. Microdialysis probes were implanted into the surgical site and dialysates, and subjective pain reports were collected every 15 minutes for 4 hours after surgery. Tissue levels of iBK were measured using a radioimmunoassay. Preoperative administration of flurbiprofen significantly reduced patients' reports of pain from 120 to 240 minutes after surgery and blocked the peak increase in tissue levels of iBK (135 to 150 minutes after surgery). Although these results indicate that flurbiprofen has an "antibradykinin" effect, the analgesia both preceded and persisted beyond the inhibition of iBK levels. Accordingly, an antibradykinin effect may only partly contribute to flurbiprofen analgesia. The data are consistent with the hypothesis that prostaglandins contribute to the peak release of iBK owing to the potent inhibition of prostaglandin synthesis by flurbiprofen, but other yet unidentified mediators are also required for the sustained release of this inflammatory mediator into the surgical field. PMID- 8426249 TI - Carbon dioxide laser vermilionectomy for actinic cheilitis. AB - Actinic cheilitis is a premalignant condition of the lower lip that requires treatment. Several methods have been advocated to manage this disease. In this study, 19 cases of actinic cheilitis were treated by carbon dioxide (CO2) laser vermilionectomy with good results. Because it has the advantages of both scalpel vermilionectomy and laser vaporization of the vermilion, CO2 laser vermilionectomy is proposed as a good method of treating actinic cheilitis. PMID- 8426250 TI - The use of the holmium laser for temporomandibular joint arthroscopic surgery. AB - Temporomandibular joint arthroscopic surgery was performed on 86 human joints with a holmium yttrium aluminum garnet (YAG) laser (Coherent, Palo Alto, CA) passed through a quartz fiber. The wattage and number of pulses needed for tissue response, the temperature produced, and the average time needed for this surgery are presented. Holmium laser technology applied to the temporomandibular joint appears to be an effective, safe, and efficient modality. PMID- 8426251 TI - Reconstruction of mandibular discontinuity defects using autogenous grafting and a mandibular reconstruction plate: a prospective evaluation of nine consecutive cases. AB - This article presents the clinical results of a prospective evaluation of nine cases of mandibular discontinuity that were reconstructed using autogenous grafting and a reconstruction plate. All patients were allowed to function immediately postsurgery. Acceptable esthetic and functional results were obtained with minimal donor and recipient site complications. All cases showed complete graft incorporation and restoration of osseous continuity. PMID- 8426252 TI - Modified condylotomy for treatment of the painful temporomandibular joint with a reducing disc. AB - Data from a pool of approximately 400 patients operated on with modified condylotomy during a 9-year period are presented. The chief findings were good relief of pain and dysfunction, and reversal of the internal derangement in a high percentage of patients. There was low morbidity and remarkably few complications. Comparison of these findings with published results of alternative surgical and nonsurgical procedures seems to favor modified condylotomy. PMID- 8426253 TI - The double-reversing Z-plasty in primary palatoplasty: operative experience and early results. AB - The double-reversing Z-plasty of Furlow for closure of the soft palate was used in 34 children with various types of cleft palate. Mean age at repair was 12.8 months. Intraoperative experience was favorable, with acceptable operating time and blood loss. Length of hospitalization averaged 1.9 days. Postoperatively, two children experienced temporary stridor, which resolved within 24 to 48 hours. One child had dehiscence of the hard palate (Von Lagenbeck repair) 4 weeks postoperatively, and three children developed small oronasal fistulae. Early speech evaluation demonstrated adequate soft palate mobility in 33 of 34 patients, with observable velopharyngeal function. Twelve children had mild velar compromise, with eight exhibiting slight nasal air escape. PMID- 8426254 TI - Pterygoid plate fracture in Le Fort I osteotomy with and without pterygoid chisel: a computed tomography scan evaluation of 58 patients. AB - The incidence of pterygoid plate fracture as determined by computed tomography (CT) scan is much higher than that determined at surgery. This observation is irrespective of whether a chisel is used to effect pterygo-maxillary separation. PMID- 8426255 TI - The effects of screw number and length on two methods of tension band plating. AB - Bovine ribs were used to evaluate the effects that screw length and number have on two types of tension bands. Five trials each were done with one, two, three, or four screws per segment, using screws 4, 8, 12, or 16 mm in length and 2-mm adaptation plates or 2-mm mini-dynamic compression plates. The fractured ribs were subjected to increasing weights until failure. With the 2-mm adaptation plate, screw length played no role in the amount of weight resisted. Increasing the number of screws per segment increased the weight resisted to up to three screws per segment, after which no additional benefit was realized (P < .001). For the 2-mm mini-dynamic compression plate, increasing screw length increased the weight resisted up to three screws per segment, after which length had no effect (P < .001). Increasing the number of screws per segment increased the weight resisted up to three screws per segment, after which no additional benefit was realized (P < .001). PMID- 8426256 TI - Rigid skeletal fixation of fractures. AB - Rigid skeletal fixation of facial fractures has evolved from the principles established in orthopedics. It has taken a long time to develop rigid internal fixation devices that provide stability combined with safety. The application of rigid skeletal fixation to the facial skeleton requires the surgeon to pay strict attention to detail, which may add a small time increment to the procedure. However, the benefits to patients of having early use of the jaws and exact placement of bony segments seem to outweigh the disadvantages. The future of this constantly developing field will almost certainly center around technologic innovations that will make the application of fixation devices easier. It is likely that future research will provide devices that are more biocompatible, and perhaps just over the horizon, devices that are bioresorbable. PMID- 8426257 TI - Treatment of mammalian bite wounds of the maxillofacial region. AB - Infected human bites may result in devastating consequences. Failure to recognize the destructive potential of infections of bone, tendons, joints, and soft tissue caused by oral microorganisms can lead to serious loss of intrinsic functions. Rapid prevention or resolution of infection by appropriate use of antibiotics and surgical treatment continues to be a therapeutic challenge. PMID- 8426258 TI - A soft-tissue mass in a maxillary extraction site. PMID- 8426259 TI - Surgical management of a protein S-deficient patient. PMID- 8426260 TI - Aneurysmal bone cyst of the maxilla: report of a case. PMID- 8426261 TI - Esophageal cancer metastatic to the preauricular region in a patient with two different primary tumors: case report and review of the literature. PMID- 8426262 TI - Healing of fractures of the coronoid process: report of cases. PMID- 8426263 TI - Probable cocaine-induced hyperthermia in an anesthetized patient: a case report. PMID- 8426264 TI - Acute intraoperative hyperthermic episode: a case report. PMID- 8426265 TI - Use of a reciprocating saw for removal of mandibular tori. PMID- 8426266 TI - Proper designation of our specialty. PMID- 8426267 TI - Judging the accuracy of advertisements. PMID- 8426268 TI - Other causes of the numb chin. PMID- 8426269 TI - Preliminary findings on the variation of serum apolipoprotein levels in neural degenerative disorders. AB - We measured six apolipoproteins (AI, AII, B, CII, CIII, and E) in the serum of patients with several kinds of neural diseases [diabetic neuropathy and neural degenerative disorders (motor neuron degenerative disorders, spinocerebellar degeneration, Parkinson's disease)], comparing them to the age-matched healthy controls using the immunoturbidimetric method. Statistically significant decreases of serum apo-AI, apo-A-II and increases of apo-CIII, apo-E were observed in neural degenerative diseases; and, particularly, higher apo-B and apo CII concentrations were observed in diabetic neuropathy. Most neural degenerative disease showed lower apo-AII. However, in motor neuron degenerative disorders, higher apo C-II and apo-E were seen. Lower apo-AI was seen in spinocerebellar degeneration, and lower apo-AII was seen in Parkinson's disease. Higher apo-B, CII, and E levels were observed in females with spinocerebellar degeneration and Parkinson's disease, and lower apo-AII was seen in males with spinocerebellar disease. PMID- 8426270 TI - Rhabdomyosarcoma cell line can be used for the isolation of soluble acetylcholine receptor and for assaying blocking and modulating autoantibodies. AB - We found that the Rhabdomyosarcoma (RD) cell line expresses human acetylcholine receptor (AChR) based on the following evidences: 1. Soluble AChR can be isolated from RD cells following the isolation procedure for AChR from human muscle; 2. Intact RD cells bind to alpha-bungarotoxin (alpha Butx) in a time-dependent and saturable fashion. The apparent dissociation constant (5.3 x 10(-10) M) is very similar to that reported for TE671 cells, which is known to express AChR; 3. Like fresh muscle culture, RD cells not only bind but also internalize 125I-alpha Butx. Soluble AChR from RD cells can be labeled specifically with 125I-alpha Butx and then used to quantify binding autoantibodies in myasthenic patients. We also demonstrate that blocking antibodies can be detected in sera from patients with myasthenia gravis (MG) using RD cells and the ability of RD cells to internalize alpha Butx. Consequently, RD cells can be used as a reliable source for obtaining soluble AChR and as a replacement for rodent or human muscle cultures in measuring blocking and modulating antibodies. PMID- 8426271 TI - Evaluation of the Hawaiian reef fishes with the solid phase immunobead assay. AB - This study presents data on the evaluation of a laboratory ciguatera kit based on the solid phase immunobead assay (SPIA) for the detection of ciguatoxin in Hawaiian reef fish. The SPIA was performed on fish catches by volunteer fishermen throughout the State of Hawaii. A total of 1,067 fish of various species were tested for ciguatoxin (CTX) using the SPIA kit. Of the total 1,067 fish tested, 510 were from Oahu, 402 from Hawaii, and 75 from Maui. The number of fish from Molokai, Kauai, and Lanai were 23, 20, and 7 respectively. Twenty percent of the total fish tested were positive, 41% borderline, and 39% negative for ciguatoxin. The highest percentage of SPIA- positive fish were from Hawaii (27%) followed by Oahu (19%) and Kauai (15%). These results correlate with the reported incidents from the Department of Health (DOH) of actual ciguatera poisoning in the State of Hawaii. Fish in all three categories of the SPIA test values were eaten. No false negatives were noted with individuals eating SPIA negative fish. Of the 232 SPIA borderline values eaten, 3 species of fish caused ciguatera poisoning. These fish included 2 papio, 1 mullet, and 1 po'ou. Of the 17 SPIA positive fish eaten, 5 caused ciguatera poisoning: 2 papio, a kole, an uhu, and a weke. The SPIA ciguatera test did protect the public when only SPIA-negative fish were eaten. The borderline and positive SPIA fish were generally unsafe, especially the positive fish. The data indicated that the probability of getting ciguatera with a SPIA positive fish was 1 out of 3.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426272 TI - Characterization of marine toxin(s) in Myripristis sp. by immunological, mouse toxicity, and guinea pig assays. AB - Myripritis sp. (squirrelfish) has been assessed for toxicity by 1) the stick enzyme immunoassay (S-EIA); 2) mouse toxicity bioassay; and 3) guinea pig atrial assay. Analysis of Myripristis flesh with MAb-CTX and MAb-OA showed that with every fish examined, the reaction with MAb-OA was considerably higher. The mean S EIA value for MAB-OA was 2.9 +/- 0.8 while the mean for MAb-CTX was 1.7 +/- 0.5. The data strongly suggests that Myripristis sp. appear to contain okadaic acid like toxins and/or mixed with CTX. Five fractions of the flesh extracts were obtained by silica gel chromatography. These included 100% CHC1(3), 10% MeOH/CHC1(3), 50% MeOH-CHC1(3), 100% MeOH, and 80% MeOH/H2O. The 100% CHC1(3) eluate proved to be the most toxic (mouse killed in 32 minutes) and the 10% MeOH CHC1(3) fraction killed in approximately 48 hours. The remaining fractions showed a significantly lower toxicity level in mice. In the guinea pig atria examinations, extracts of Myripristis flesh, gut, and crustacea (from the gut) were studied. Extracts of the gut and crustacea showed strong sodium channel blockage, while the flesh extracts showed a weak inotropic effect, characteristic of okadaic acid. The latter response was only blocked by verapamil, but not with tetrodotoxin (10(-5) M) or the adrenergic blockers (10(-5)M). The data from this study suggest toxic compound(s) not previously reported, in the non-polar fraction of Myripristis flesh having a sodium channel blockage effect. PMID- 8426273 TI - Stability of respiratory syncytial virus antigen due to buffer treatment for direct detection in nasopharyngeal specimens with enzyme immunoassay. AB - We developed an enzyme immunoassay (direct EIA; Enzygnost RSV[Ag]) for the direct detection of respiratory syncytial virus (RSV) antigen in nasopharyngeal specimens (NPS). The test procedure is the same as our recently described direct EIA for detection of influenza A and B virus antigens in NPS. For practical purposes it is of advantage to differentiate respiratory viruses on the same microtitration plate in the same run. The test shows no limitations by sample consistency, and results are obtained within 4 hr. In contrast to other test systems, sonification is not necessary. This is due to the sample buffer STD. We studied the influence of sample buffer STD on the stability of RSV (strain Long) antigen at different temperatures over a period of 7 days. PBS-BSA-buffer served as control. The treatment and storage of RSV (strain Long) with sample buffer STD at room temperature or at 4 degrees C showed no decrease of antigen detectability. The antigen is very stable in contrast to the storage of RSV (strain Long) in PBS-BSA buffer during the observation period of 7 days. Consequently, when NPS are stored in sample buffer STD, results of direct EIA are independent from the time of transport and temperature within 7 days. Thirty eight NPS from infants with confirmed RSV infection were investigated. Confirmation was performed by virus isolation (n = 29) or with commercially available enzyme immunoassays or immunofluorescence test (n = 9). The direct EIA showed a specificity of 99.3% (n = 140) and a sensitivity of 95% (n = 38). PMID- 8426274 TI - Autoantibodies in the sera of Trypanosoma cruzi-infected individuals with or without clinical Chagas disease. AB - Variations in the levels and the specificities of autoantibodies directed against a panel of antigens (cytoskeleton proteins, DNA, laminin) were analyzed in the sera from two groups of humans infected with Trypanosoma cruzi. One group was constituted of apparently healthy blood donors (BD) and the other of patients with clinically confirmed Chagas disease (CCH). In both infected groups, a high proportion but not all sera exhibited dramatic enhancement of IgM and IgG autoantibodies directed against all antigens tested. Sera positive for IgG autoantibodies were generally found more frequently in the CCH than in the BD group, except for anti-actin antibodies more often present in BD sera. Anti laminin IgG antibodies were present in a similar number of individuals in both groups. Although the titers of anti-laminin IgG antibodies were in general higher in CCH, their dissociation constants were in the same range (7 x 10(-8) - 10(-7) M) in both groups. IgG autoantibodies were demonstrated to be polyreactive with laminin and other self antigens as well. Circulating immune complexes were present in sera from both groups and the activity of the antibodies dissociated from these complexes was directed against all the antigens of the panel. Although the IgE concentration was significantly enhanced in several subjects from both groups, the incidence of positive sera was higher in the CCH (60%) than in the BD (39%) group. Our results demonstrate that autoantibodies with the characteristics of natural autoantibodies are found in both T. cruzi-infected apparently healthy individuals and patients. PMID- 8426275 TI - Measurement of anti-thyroglobulin IgG in urine of patients with autoimmune thyroid diseases by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay). AB - Anti-thyroglobulin IgG in urine of patients with Graves' disease and chronic thyroiditis and healthy subjects was measured by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay). Anti-thyroglobulin IgG in dialyzed urine was reacted simultaneously with 2,4-dinitrophenylated thyroglobulin and thyroglobulin-beta-D-galactosidase conjugate. The immune complex formed consisting of the three components was trapped onto polystyrene balls coated with (anti-2,4-dinitrophenyl group) IgG, eluted with epsilon N-2,4-dinitrophenyl-L lysine, and transferred onto polystyrene balls coated with (anti-human IgG gamma chain) IgG. beta-D-Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. Anti-thyroglobulin IgG was detected in most of the patients, but not in most of the healthy subjects; levels of anti-thyroglobulin IgG in urine of the patients were well correlated to those in serum of the same patients. The measurement of anti-thyroglobulin IgG in urine by the immune complex transfer enzyme immunoassay was suggested to be useful as a diagnostic aid for autoimmune thyroid diseases. The conventional standard ELISA was not sufficiently sensitive for measuring anti-thyroglobulin IgG in urine. PMID- 8426276 TI - Comments on "Comparison of two automated nephelometers" by Emancipator et al. PMID- 8426277 TI - Extracellular matrix analysis of nifedipine-induced gingival overgrowth: immunohistochemical distribution of different collagen types as well as the glycoprotein fibronectin. AB - The purpose of this study was to demonstrate the localization of collagen types I, III, IV, V, VI and VII as well as the glycoprotein fibronectin in nifedipine induced gingival overgrowth. The slices, after the use of indirect immunofluorescence (incubation with antibodies against these extracellular matrix components), showed a diffuse distribution with the anti-types I and III in the stroma and fluorescent staining of the basement membranes of the epithelium, blood vessels and nerves with collagen type IV antibodies. The increased number of vessels was localized near the surface of the lesion. Collagen type V - seen as a filamentous - and collagen type VI - as microfibrillar - components were also localized in the tissue, showing completely different patterns of distribution. Collagen type V appeared "crater"-like and type VI displayed a "honeycomb"-shaped structural model. The blood vessels were not stained but the area around their walls demonstrated an intense fluorescence with these antibodies. Collagen type VII showed a characteristic linear staining near to the epithelial basement membrane. In contrast to this, fibronectin localized with a varied intensity in the different areas of the tissues and presented a "cloud" like structure. This shows differences between the matrix components in nifedipine-induced hyperplasia and confirms the heterogeneity of the matrix in health and in gingival alterations. PMID- 8426278 TI - ELISA detection of glycosaminoglycan (GAG)-linked proteoglycans in gingival crevicular fluid. AB - The purpose of this study was to develop an ELISA method to detect chondroitin sulfate isomer-linked proteoglycans in gingival crevicular fluid (GCF), and to elucidate the role played by the glycosaminoglycans (GAGs) in GCF during experimentally-induced periodontitis in dogs. Experimental periodontitis was induced by placement of a silk ligature below the gingival margin of the molar teeth in 3 mongrel dogs. GCF was collected using microcapillary tubes at 0, 7, 21 and 60 days after ligature placement. To compare with GAG in GCF, bovine nasal cartilage proteoglycan monomer, dog's serum and supernatant of homogenized gingival tissue were prepared. Combination of monoclonal antibodies, 3B3 and 9A2, and specific enzymatic digestion made possible the identification of chondroitin 4 sulfate (C4S), chondroitin 6 sulfate (C6S) and dermatan sulfate (DS). The ELISA method detected very small amount of chondroitin sulfate (CS) isomers (15-1000 ng/ml of bovine nasal cartilage proteoglycan). The ELISA value of CS isomers in GCF was lower than that of homogenized gingival tissue but higher than that of the serum. The ELISA value of C4S, C6S and DS, although fluctuating, increased in proportion to the severity of the inflammation. PMID- 8426279 TI - Purification of arginine-sensitive hemagglutinin from Fusobacterium nucleatum and its role in coaggregation. AB - Hemagglutinin of Fusobacterium nucleatum was extracted from Triton X-100-pronase P-treated cell envelopes, and was purified by affinity chromatography on L arginine agarose. The hemagglutinin was inactivated by heating at 70 degrees C for 1 min. The activity was inhibited by L-arginine but was not affected by any sugars or by EDTA. The hemagglutinin aggregated 14 out of 17 strains of oral streptococci tested, and the bacterial aggregating activity was also inhibited by L-arginine. The results indicate the dominant role of this hemagglutinin in the adherence of this bacterium both to host cells and to other bacteria. PMID- 8426280 TI - Localization of interleukin-1 (IL-1) mRNA-expressing macrophages in human inflamed gingiva and IL-1 activity in gingival crevicular fluid. AB - The exact cell type and site(s) involved in interleukin-1 (IL-1) production during gingival inflammation was determined by combining immunohistochemistry and in situ hybridization. IL-1 messenger RNA (mRNA)-expressing cells in human inflamed gingiva were identified as macrophages. The rate of IL-alpha mRNA expression in these macrophages was the same as IL-1 beta mRNA expression. The rate of IL-1 mRNA expression was higher in connective tissue furthest from the pocket epithelium, although more macrophages were present at the connective tissue subjacent to the pocket epithelium. The IL-1 activity in gingival crevicular fluid (GCF) obtained from inflamed gingiva was higher than that from healthy gingiva and decreased after periodontal therapy. The IL-1 activity in GCF was almost completely abolished by the addition of anti-IL-1 alpha antibody but not by anti-IL-1 beta antibody, indicating that IL-1 alpha is the predominant form in GCF. However, the IL-1 activity in GCF was unrelated to the number of IL 1 mRNA-expressing macrophages in the same gingival site where the GCF was obtained at the same time. The results suggest that macrophages in the connective tissue subjacent to the oral epithelium contribute to the production of IL-1 but those in connective tissue subjacent to the pocket epithelium play a different role in the generation of gingival inflammation. PMID- 8426281 TI - Protein, albumin and cystatin concentrations in saliva of healthy subjects and of patients with gingivitis or periodontitis. AB - Salivary protein, albumin and cystatin concentrations were investigated in subjects with a healthy periodontium and in patients with gingivitis or periodontitis. Protein and albumin concentrations in saliva of individuals with gingivitis or periodontitis were significantly increased compared with healthy subjects. Salivary protein and albumin appeared to be positively correlated in all the groups, which suggests that the increase in salivary protein concentration in subjects with gingivitis or periodontitis is caused by leakage of plasma proteins. Cystatin concentrations in saliva of subjects with periodontitis were significantly increased when compared with the healthy group and the gingivitis group (p < 0.01). In the gingivitis and periodontitis group, salivary cystatin was only weakly correlated with albumin concentrations, which suggests that the increased salivary cystatin activity found in subjects with gingivitis and periodontitis is derived from sources other than plasma. PMID- 8426282 TI - Multinucleated fibroblastic cells in the periodontal ligaments of aged rats. AB - Using 12- to 18-month-old rats, we examined the ultrastructural and cytochemical features of multinucleated fibroblastic cells (MFCs) in the periodontal ligament (PDL) of molars. In aged rats, the MFCs were distributed randomly in the PDL and exhibited cytoplasmic structural variations which were not dependent on the number of nuclei. There was a tendency for the MFCs to cluster in the PDL. The MFCs, rich in cytoplasmic organelles involved with procollagen synthesis such as rough endoplasmic reticulum and the Golgi apparatus, incorporated and secreted 3H proline-labelled products. The MFCs also possessed many phagosomes containing intact collagen fibrils. These MFCs were apparently involved in phagocytosis and intracellular degradation of incorporated collagen fibrils. Phagosome-rich MFCs contain acid phosphatase activity in primary and secondary lysosomes, similar or stronger in intensity to that which can be demonstrated in mononuclear fibroblasts. However, unlike mononuclear fibroblasts, the MFCs did not exhibit alkaline phosphatase activity along their plasma membranes. These results suggest that MFCs demonstrate a range of fibroblastic cellular activity, including collagen phagocytosis, and that they may lack certain plasma membrane glycoproteins, which might explain the occurrence of multinucleation in these cells. PMID- 8426284 TI - The role of metronidazole in the treatment of periodontal diseases. AB - This article discusses the ability of metronidazole to improve periodontal health. Review of the drug's pharmacology and potential side effects indicate that it poses little threat to humans of inducing acute toxicity, mutagenesis, or cancer if used according to recommended dosing regimens. Studies addressing metronidazole utilization in a variety of clinical conditions demonstrate that its routine use does not enhance root planing. However, adjunctive antibiotic therapy may be advantageous in the treatment of sites where effective root planing is precluded due to deep pockets or when anaerobic periodontal infections do not respond to conventional therapy. PMID- 8426283 TI - Sub-gingival microflora in Macaca mulatta species of rhesus monkey. AB - The Macaca mulatta species of rhesus monkey is one of several non-human primate (nhp) models for periodontal disease. This report presents the bacteriology of the gingival sulci in M. mulatta monkeys. Three sub-gingival sites (maxillary right central incisor, the disto-buccal of the mandibular left second molar and mesio-buccal of the mandibular right second molar) of 9 monkeys were evaluated clinically before scaling and 7 days after scaling. Plaque samples were obtained from sub-gingival sites before clinical examination and studied bacteriologically by dark field microscopy, selective and non-selective culture, and by primary phenotypic characterizations of culture isolates. Several gingival sites presented with mild gingival inflammation. Anaerobic and facultatively anaerobic bacteria were the predominant flora colonizing the gingival sulci. The major microbial groups were Haemophilus species (100% of sites; percentage of total anaerobic count (TAC): 21-51), Peptostreptococcus micros (89%, 7.5-29.5), Actinomyces sp. (85%, 7-27), Fusobacterium nucleatum (90%, 5-8), Actinobacillus actinomycetemcomitans (73%, 1.3-12), black-pigmented anaerobic rods (BPAR) (80%, 0.6-6.5) and oral streptococci (80%, 0.2-1.0). Microbial groups detected less often were Wolinella sp. (66%, 0-2.6), Capnocytophaga sp. (30%), Eikenella corrodens (4.7%, 0), Campylobacter sp. (28%, 0-0.1) and spirochetes (4.7%, 0 0.07). Seven days after gingival sites were scaled, the plaque score and indices for gingival inflammation declined significantly. The gingival flora after scaling were characterized by lower proportions of the Actinomyces sp., P. micros and BPAR; and increased proportions of the oral streptococci, relative to pre scaling levels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426285 TI - Evidence for cigarette smoking as a major risk factor for periodontitis. AB - The role of smoking as a risk factor for periodontitis was assessed separately in diabetic and nondiabetic study groups. Subject listings stratified for age (19 to 40 years) and sex were obtained for subjects with insulin-dependent diabetes mellitus (IDDM) and nondiabetic subjects. For both the IDDM group (n = 132) and the nondiabetic group (n = 95), age and sex stratified samples were constructed by random selection of subjects from each subject listing. Patients were recruited by phone, examined, and their medical and dental histories obtained. Among nondiabetic subjects, the prevalence of periodontitis was markedly higher among current smokers compared with never smokers (P < or = 0.005) in both the 19 to 30 year-old (46% vs. 12%) and 31 to 40 year-old groups (88% vs. 33%). The subject mean percent of sites with gingival pocket depth > or = 4 mm was higher among current smokers than never smokers (P = 0.001) in the 19 to 30 (8.2% vs. 3.4%) and 31 to 40 (14.3% vs. 4.3%) age groups. The effects of smoking among IDDM subjects were similar to that observed in the nondiabetic population. There were no differences between current and never smokers in the proportion of sites positive for plaque. Attributable risk percents from prevalence data suggest that among nondiabetic subjects, a large proportion, perhaps as much as 51% of the periodontitis in the 19 to 30 year old group and 32% of the periodontitis in the 31 to 40 year old group, is associated with smoking.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426286 TI - Short-term effect of cyclosporin injections on experimental gingival inflammation in the rat. AB - Experimental gingivitis was induced in 2 groups of 10 adult Wistar rats by feeding them a high sucrose diet for 30 days. Since cyclosporin has an immunosuppressive effect on the stimulation of T lymphocytes, the test group received 5 intraperitoneal injections of cyclosporin A from day 20 to 30 while the control group received no additional treatment. The gingival regions of the molars were studied in light and transmission electron microscopy. In both groups, a well-differentiated wall of polymorphonuclear neutrophils (PMNs) was present between the dental plaque and the papillary epithelium and no bacterial invasion occurred despite the local absence of T lymphocytes in the test group. It can be concluded that T lymphocytes are not involved in the mechanism of bacterial invasion. PMID- 8426287 TI - Periodontal ligament formation around different types of dental titanium implants. I. The self-tapping screw type implant system. AB - The aim of this study was to determine if a periodontal ligament can form around self-tapping, screw type titanium dental implants. Implants were inserted in contact with the periodontal ligament of root tips retained in the mandibular jaws of 7 monkeys. In each side of the mandible, 1 premolar and 2 molars were removed in such a manner that in approximately half the cases, the root tips were retained. Following healing, the experimental areas were examined on radiographs, and sites were selected for the insertion of the implants, so that every second implant would have a close contact to the retained root tips. Three months after placement of the implants, the animals were sacrificed, and ground sections were produced for histological analysis. The results demonstrated that newly-formed cementum had become deposited on the implant surface in the contact area between the implant and the retained roots, whereas osseointegration was consistently found on the remaining part of the implant surface as well as on the entire surface of the implants placed without contact to the retained root tips. The study has shown that, when placed in contact with periodontal ligament tissue of retained roots, a periodontal ligament can form on self-tapping, screw type titanium dental implants in areas where a void is present between the surrounding bone and the implant at the time of insertion. PMID- 8426288 TI - Clinical evaluation of a constant force electronic probe. AB - This study aimed to compare, in vivo, a conventional pocket probe with an automatic, computerized, constant force, electronic probe with a discrimination ability up to 0.1 mm. Sixteen adults with moderate chronic periodontitis and free of supra- and subgingival calculus participated in this study. Eight patients were examined by 2 investigators who used both the conventional and the automatic probes, for a total of 4 probings per subject. The remaining 8 patients were examined 4 times by investigator 1, twice with each probe. For each patient the Ramfjord teeth were examined and 6 sites were considered per tooth. Although the pocket depth measurements recorded by the manual probe were consistently deeper than those of the electronic probe, a good correlation was found between both recordings. Moreover, intra- and inter-examiner comparisons showed comparable standard deviations for both probes and small differences in absolute scores. The conventional probe was slightly more reproducible whereas the automated probe had the advantage of automatic registration. The results indicate that both probes can be considered as valuable in clinical practice. PMID- 8426289 TI - Periodontal microflora of HIV positive subjects with gingivitis or adult periodontitis. AB - The subgingival microflora of 39 HIV+ subjects with gingivitis or adult periodontitis was cultured quantitatively anaerobically for bacteria, spirochetes, and mycoplasma and aerobically for yeasts. Isolates were characterized by conventional biochemical tests, polyacrylamide gel electrophoresis of soluble proteins, cellular fatty acid profiles, immunofluorescence, and immunodiffusion. In general, the same types of bacteria were isolated from the subgingival crevice of HIV+ subjects as we previously had isolated from the subgingival crevice of non-HIV subjects. A statistically significant difference was found between the composition of the flora of HIV+ subjects with adult periodontitis (AP) and concurrent studies of a non-HIV+ AP population. Mycoplasma salivarium was significantly elevated in the HIV+ subjects examined. Yeasts were isolated from only 10% of the samples and from 13% of the HIV-positive subjects at 0.05 to 0.0002% of the total cultivable count when present. PMID- 8426290 TI - The effects of guided tissue regeneration membrane placement on healthy periodontal sites. AB - This study investigates the effects of the placement of a bioresorbable Type I collagen barrier on clinically healthy sites. Ten human subjects, with at least one pair of matched periodontal defects included in a previous study, were investigated for the effect of guided tissue regeneration (GTR) membrane placement on adjacent healthy sites. Control sites consisted of open flap debridement, while experimental sites consisted of similar debridement procedures with placement of a collagen membrane for GTR. The membrane placement was designed to completely cover the periodontal defect, and therefore of necessity overlapped adjacent periodontally healthy sites. It is these healthy sites that are included in the present study. Standardized measurements of change in probing attachment levels were obtained at the time of surgery and 1 year later. The differences in change of clinical attachment levels (CAL) were compared utilizing the Student t-test for paired samples. The mean loss of CAL in the control sites was 1.00 +/- 1.179 mm, and in the test sites was 0.60 +/- 1.729 mm. When comparing the difference in changes of CAL in both test and control sites, there was no statistical significance. There was, however, a wide range of changes between individual sites, which might suggest a clinically meaningful change of CAL on an individual tooth basis. The results of this study suggest that the placement of a bioresorbable collagen barrier has no significant effect on CAL in healthy areas. PMID- 8426291 TI - Periodontal infection leading to periostitis ossificans ("Garre's osteomyelitis") of the mandible. Report of a case. AB - Periostitis ossifications ("Garre's osteomyelitis") of the mandible is a rather rare pathology that occurs as a hard swelling at the mandibular angle, persists for a long period, and is mostly painful on palpation. Clinical signs of infection are rarely noted. The etiological factor is generally a carious first permanent molar with a periapical infection or a fracture of the jaw. In this case, a 10 year-old girl showed a fixed painful swelling of the right mandibular angle. This revealed to be periostitis ossificans, although none of the classical causes were present. The symptoms were evoked by a periodontal infection: there was an osseous crater and a probing pocket depth of more than 10 mm disto lingually of the first right permanent molar of the mandible. This was likely the etiological factor. Extraction of the first and second permanent right molar, curettage of the corresponding alveoli, and antibiotic treatment resulted in complete healing, clinically and radiologically, after 9 months. PMID- 8426292 TI - Focal epithelial hyperplasia (Heck's disease) with generalized lesions of the gingiva. A case report. AB - Focal epithelial hyperplasia is an uncommon benign oral condition that occurs mainly in young individuals of certain racial groups. The lesions most commonly affect the mucosal surfaces of the lips and they have been observed frequently to regress within a few months. This is a report of an unusual case of focal epithelial hyperplasia in a young black adult presenting with generalized involvement of the gingiva. There is no clinical evidence of regression of the lesions and the case remains similar to the original appearance 2 years after diagnosis. The diagnosis and management of focal epithelial hyperplasia are discussed. PMID- 8426293 TI - Histologic assessment of a contiguous autogenous transplant in a human intrabony defect. A case report. AB - Increased blood supply, vital bone marrow cells, and minimal mobility may play a significant role in the success of osseous grafts, and are characteristics of the bone swaging grafting technique. As in all autogenous grafts, the risk of disease transmission is minimal. Previous reports of clinical success raise questions as to the type of tissue response to this procedure. This case report examines 8 months radiographic and histologic results of a clinically successful bone swaging graft. PMID- 8426294 TI - Treatment of the palato-gingival groove with guided tissue regeneration. Report of 10 cases. AB - The purpose of this study is to demonstrate the potential of using a barrier in the treatment of palato-gingival groove defects. The study group consisted of 10 patients. Prior to treatment, the palato-gingival groove on maxillary lateral incisors was measured with calibrated periodontal probe from the cemento-enamel junction (CEJ) to the free gingival margin (FGM) and from the FGM to the base of the pocket (BP). Probing depth (PD) was calculated and bleeding on probing indicated. Surgical procedures consisted of flap reflection, removal of granulation tissue, and scaling and root planning of the groove. An expanded polytetrafluoroethylene membrane was sutured over the palato-gingival groove. Six months postsurgery, all measurements were repeated. Statistical analysis compared results using means, standard deviations, and paired t tests. Results showed an improvement in clinical attachment gain, probing depth reduction, and decreased bleeding on probing. This study demonstrates the potential of guided tissue regeneration in the treatment of palato-gingival groove defects. A random blinded clinical trial is necessary, however, to fully assess the potential of this procedure in treatment of palato-gingival groove defects. PMID- 8426296 TI - Enteropathy in HIV infection. AB - Enteropathy occurs frequently in persons infected with human immunodeficiency virus (HIV), and symptoms related to the gut are a major cause of debility. The pathogens associated with disease are diverse, often difficult to detect, and frequently poorly responsive to any therapeutic intervention. This brief review examines HIV-related enteropathy from the perspective of symptomatology, and discusses some of the recent advances in diagnosis of specific gut disorders. PMID- 8426295 TI - Epidemiological pattern of HIV infection in Australia. AB - Human immunodeficiency virus (HIV) infection in Australia has been transmitted primarily through sex between men. Multiple sources of information, including backprojection from AIDS cases and longitudinal studies involving repeat testing for HIV antibody, indicate that the rate of HIV transmission through sex between men reached a peak in 1983-1984 and declined rapidly in subsequent years. However, there is also evidence that HIV infection continues to be transmitted through sex between men in Australia. Intravenous drug use has been reported by only a small proportion of people diagnosed with HIV infection in Australia, and surveys of intravenous drug users have found a low rate of infection apart from in men who had homosexual contract. Behavioral studies have found that sharing of injecting equipment has continued, suggesting that low infection rates may be a consequence of sharing patterns. Heterosexual transmission seems to have been infrequent in Australia, perhaps because of the low rate of HIV infection in intravenous drug users. PMID- 8426298 TI - HIV antibody testing in Australia. AB - Testing for antibody to human immunodeficiency virus (HIV) in Australia is subjected to continual monitoring for ensuring that the performance of tests in the field is reliable and provides accurate data on HIV tests and testing that are unique to Australia. PMID- 8426297 TI - Pediatric HIV: Australian perspective. AB - Pediatric infection with human immunodeficiency virus (HIV) in Australia, as elsewhere, now reflects HIV seroprevalence in women of childbearing age although numbers remain small. A perinatal transmission rate of approximately 30% has been observed. The rate appears to be higher for breast-fed babies; exclusively bottle fed babies of HIV-infected mothers appear to have a risk of < 20% of acquiring HIV perinatally. Although the seroprevalence rate in women of childbearing age in Australia remains low, routine antenatal screening will not be cost-effective. Although passively acquired maternal antibody may confound serodiagnosis for up to 18 months, in practice, HIV infection, when present, can be detected clinically or with use of other laboratory parameters (p24, polymerase chain reaction, viral culture, hyperglobulinemia) during the first 6 months of life. Zidovudine appears to be relatively nontoxic in children and effective in producing weight gain and preventing opportunistic infection. Although experience with didanosine is limited, this drug appears to have a very acceptable toxicity profile and the advantage of twice-daily dosing. However, the bioavailability and penetration into the cerebrospinal fluid are poor. Neurological disease can improve when absorption is adequate. Residential family camps may help to reduce isolation for families and provide them with an opportunity for improving their understanding of HIV through interaction with peers and professionals. PMID- 8426299 TI - Clinical immunology and HIV infection. AB - Clinical immunologists play a major role in the multidisciplinary approach to the management of human immunodeficiency virus (HIV) infection and its complications in Australia, through involvement with the clinical and laboratory management of patients as well as with research on disease pathogenic mechanisms. Consequently, immunological monitoring of the pathogenic effects of chronic HIV infection is commonplace, allowing for studies on the clinical utility of immunological investigations as a means of monitoring the severity of HIV-induced immunopathology and immunodeficiency to be undertaken. Such studies have demonstrated the clinical utility of CD4+ T-cell assays, but studies on cutaneous delayed-type hypersensitivity have demonstrated that the cellular immunodeficiency induced by chronic HIV infection may be more complex than CD4+ T cell depletion alone. In addition, the involvement of clinical immunologists has greatly facilitated studies on the pathogenic mechanisms underlying HIV-induced immunopathology. Such activities place Australia in a very advantageous position for future studies on the use of immune-based therapies for controlling the pathogenic effects of chronic HIV infection. PMID- 8426300 TI - Viral markers in HIV infection and AIDS. AB - Viral and immune markers are used for monitoring either progression of human immunodeficiency virus (HIV) disease or response to antiviral therapy. Ideal properties of viral markers are that they are present in all HIV-infected persons at all stages of disease, that they are related to disease pathogenesis, that they can be easily quantitated, that this quantitation correlates rapidly and predictably with both disease stage and response to antivirals, and that they can be developed into rapid, reproducible automated tests. Currently available viral markers include HIV p24 antigenemia (after acid glycine dissociation), anti-p24 antibody titres, quantitative DNA and RNA polymerase chain reaction performed on cells and plasma, and HIV isolate phenotype. In Australia, these markers have been studied in acute HIV seroconversion, in neonatal infection, in body fluids other than blood, and in monitoring of response to antiviral drug therapy. PMID- 8426301 TI - New developments in the clinical use of didanosine. AB - Didanosine (ddI) is a purine analogue that demonstrates in vitro anti-human immunodeficiency virus (HIV) activity, effects on the surrogate markers CD4+ T lymphocytes and p24 antigen, and has adequate oral bio-availability. Recently, 500 mg/day of ddI in sachet form demonstrated clinical effectiveness compared with zidovudine (ZDV) for delaying HIV disease progression in patients with AIDS or AIDS related complex (ARC) and less than 300 CD4+ T-lymphocytes, or asymptomatic individuals with less than 200 CD4+ T-lymphocytes. All patients tolerated a minimum of 16 weeks of prior ZDV treatment. ddI treatment was associated with an increase in serum amylase and pancreatitis, however, there was no significant difference in the incidence of pancreatitis between the 500 mg ddI and ZDV groups. There was significantly more hematologic toxicity associated with ZDV and no difference between ddI and ZDV groups with respect to peripheral neuropathy. ddI is presently available in tablet form with 125% the bioavailability of the sachet form of ddI; therefore, the 500-mg sachet formulation corresponds to a 400-mg daily tablet dose of ddI. Future studies of ddI will involve ddI's effects on antiretroviral naive patients and the potential of combining ddI with other agents. PMID- 8426302 TI - HIV and malignancy. AB - Although a number of tumors in patients infected with the human immunodeficiency virus have been reported, Kaposi's sarcoma and non-Hodgkin's lymphoma remain the two principal malignancies. This article provides an Australian perspective of the recent advances in research into malignancy in HIV-infected persons. PMID- 8426303 TI - Prophylaxis of opportunistic infections in patients with HIV infection. AB - Prophylaxis of opportunistic infections in patients with human immunodeficiency virus (HIV) infection has been one of the major factors responsible for increasing patient survival. Secondary prophylaxis of most opportunistic infections is warranted, although in most instances the optimal therapy remains to be determined. Primary prophylaxis of opportunistic infections is an attractive concept, as the majority of opportunistic infections are reactivations of latent infections. Primary prophylaxis against Pneumocystis carinii pneumonia is both effective and necessary, but the role for primary prophylaxis against other infections is as yet unclear. Use of prophylactic agents requires careful study for determining the most effective and least toxic forms of prophylaxis that will allow for adequate concurrent antiretroviral therapy but will not induce resistant organisms. PMID- 8426304 TI - Immune modulation in HIV infection: fact or fantasy? AB - Since the recognition of AIDS a decade ago, many have postulated that immune modulators would provide the therapeutic intervention necessary for preventing disease progression and death. However, despite rapid advances in our understanding of disease pathogenesis and significant achievements in the area of antiviral development, immune modulators remain largely the subject matter of philosophical editorials rather than of definitive efficacy studies. More than 50 agents have been examined to date, yet only the colony-stimulating factors have proved useful in neutropenic patients with infection with human immunodeficiency virus (HIV). Other agents, such as isoprinosine, have provided conflicting data, whereas immunosuppressive agents, such as pentoxifylline, are presently under study. Cytokines examined prior to the development of antiviral agents were disappointing. Further studies examining these cytokines in conjunction with antiviral agents may be more rewarding. Other applications of these agents may be as adjuvants with vaccine administration or antimicrobial agents in the setting of opportunistic infection. To date, however, the data would suggest that there will be greater advantages from antiviral agents that prevent the development of immunosuppression than with immunopotentiators, which try to boost the embattled immune system. PMID- 8426305 TI - Current status of clinical trials in HIV disease in Australia. AB - Clinical trials in Australia in diseases related to human immunodeficiency virus (HIV) have steadily expanded from the earliest national study of zidovudine to a range of trials either actively recruiting or being activated that cover a wide spectrum of primary viral, opportunistic infection and opportunistic malignant diseases. From its earliest establishment as the National Health and Medical Research Council special unit to its current role as the clinical trials unit of the National Centre in HIV, this central group has organized a coordinated approach to the conduct of clinical trials that has been a hallmark of Australia's response to the treatment of HIV-related disease. The original local trials were in the form of collaboration in multicenter trials, initiated by the pharmaceutical industry, with the participation of only a few major treatment centers. This has now evolved to a network of more than 20 hospital and ambulatory care centers and a group of enthusiastic and committed general practitioners who have contributed data to a number of major studies. These include the national zidovudine study that generated data on the use of surrogate markers such as CD4 and p24 antigen as predictors of survival, studies of the combination of acyclovir and zidovudine that have indicated a new direction in combination antiretroviral therapy, placebo-controlled studies of zidovudine in asymptomatic individuals that may alter the timing of introduction of therapy, and the MRC/INSERM study of didanosine in advanced disease that will give the clearest indication of its activity yet to be reported.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426306 TI - Developing quality primary care services in HIV/AIDS care: the educational imperative. AB - In recognition of the growing demand for services related to infection with human immunodeficiency virus (HIV) in nonmetropolitan centers, the New South Wales Department of Health has developed a program for training selected medical practitioners from throughout the state in the management of HIV infection. This article provides a summary of the concerns and issues related to developing "A Short Course in HIV Medicine" for medical practitioners. This course was designated a prerequisite for the authority to prescribe zidovudine outside teaching hospitals and specialized HIV infection units. From our experience, we believe that an interactive approach is required in teaching, with a flexible course structure that allows the varying needs of medical practitioners with differing experiences in HIV infection to be met. From our observations we believe that additional effort is required to ensure the ongoing training of participants and to encourage greater networking with specialized HIV services. PMID- 8426307 TI - The role of the primary care physician. AB - Increased emphasis on community-based care of people with human immunodeficiency virus (HIV) infection and AIDS is recommended by the National HIV/AIDS Strategy. Patients prefer to be treated within their community, and primary care reduces the demand on scarce, costly, and already overloaded hospital facilities. Thus, there is an increasingly large role for primary care physicians to play in the management of people infected with HIV. The traditional primary health care approach of health promotion and disease prevention that focuses on case-finding, continuity of care and problem resolution, adapts well to HIV/AIDS. Primary care is holistic, patient based, and has as its focus healing rather than cure. Primary care physicians have a role in the prevention of HIV infection, in identifying asymptomatic seropositive people, in offering early therapeutic interventions, in the early detection of opportunistic infections and HIV-related malignancies, and in the ongoing management of chronic ill-health. For most people, the primary care sphere is the appropriate level for palliative and terminal care. There is also a role for primary care physicians in the psychosocial management of people with HIV/AIDS in supporting those close to the patient, and in educating the community in general about the social parameters of HIV/AIDS. PMID- 8426308 TI - Capsular epithelium: a consequence of cataract surgery. PMID- 8426309 TI - Consultation section. Antibacterial prophylaxis. PMID- 8426310 TI - Glare disability. PMID- 8426311 TI - Structural features of intraocular lenses designed for use after capsulorhexis. PMID- 8426313 TI - Anterior capsulorhexis in hypermature cataracts. PMID- 8426312 TI - Cataract surgery without retrobulbar or peribulbar anesthesia. PMID- 8426314 TI - Evaluation of a one-piece poly(methyl methacrylate) intraocular lens with a 7 mm biconvex optic and a total diameter of 10 mm. AB - To find out how large the total diameter of an open-loop intraocular lens (IOL) should be, we studied 90 consecutive implantations of a one-piece poly(methyl methacrylate) IOL with a 7 mm biconvex optic and a total diameter of 10 mm (Adatomed 75 ST). The lens design made implantation simple, even through a small capsulorhexis. In 85% of the cases, the IOL remained well centered. One case with asymmetric placement of the IOL showed a decentration of more than 1 mm. In 14% of the eyes slight decentrations occurred because the capsular bag was too large. Contact between the iris and IOL was found in 15%. In 4% there were folds in the posterior capsule, and in 35% there was a small space between the IOL and the posterior capsule because the capsular bag was too large. Although the ST lens has a small total diameter, it remained at a distance from the iris in most cases. In-the-bag implantation through a continuous central capsulorhexis is mandatory to avoid secondary displacement; nevertheless, slight decentrations cannot be excluded. For a large capsular bag, the total diameter of an IOL should exceed 10 mm. Clinical results, cellular reaction on the lens surface (26.6%), biocompatibility, and visual performance are comparable to those of other well approved poly(methyl methacrylate) IOLs. PMID- 8426315 TI - Contrast sensitivity in multifocal intraocular lenses. AB - Contrast sensitivity was measured in two groups of 20 patients each implanted with refractive and diffractive multifocal intraocular lenses and in two control groups of 20 patients each--the first group implanted with a monofocal IOL and the second phakic subjects. All cases had a postoperative follow-up of at least one year and a corrected visual acuity of 20/20- or better. We used two psychophysical tests, Pelli Robson test chart and Vistech 6500 test chart, and an objective test, visual evoked potentials (VEPs). There were no statistically significant differences in contrast sensitivity in the psychophysical tests between the two groups implanted with multifocal IOLs. The situation was different, however, when they were compared with the control group with monofocal IOLs and the group with phakic eyes: the Pelli-Robson test results were not significantly different, but the Vistech 6500 test showed a significant reduction in contrast sensitivity in both groups. The pattern VEPs objective test confirmed these results: no differences were noted between the two different multifocal IOLs, while there was a drop in contrast sensitivity when their results were compared with those of the control groups; the intermediate frequencies were particularly affected by this phenomenon. The contrast sensitivity in patients with multifocal IOLs is reduced despite high visual acuity and this can affect the quality of vision. PMID- 8426316 TI - Prospectively randomized trial comparing the pseudoaccommodation of the AMO ARRAY multifocal lens and a monofocal lens. AB - A prospective trial was set up to compare pseudoaccommodation. Fifty patients were randomized into two groups. Group 1 comprised 25 patients who had a monofocal implant and a postoperative refractive aim of -0.3 to -1.3 diopter (D) spherical equivalent and an astigmatic component between 1.0 and 1.75 D cylinder. Group 2 comprised 25 patients who had a new multizone refraction style multifocal implant and a postoperative refractive aim of -0.5 to +0.5 D spherical equivalent and an astigmatic component of less than 1.0 D cylinder. Eighty percent of postoperative refractions in Group 1 and 88% in Group 2 were within 0.5 D either side of the refractive aim. Forty-eight percent of Group 1 and 72% of Group 2 could see both 20/40 and J3 unaided. Four percent of Group 1 and 84% of Group 2 could read J2 with the distance correction. Sixteen percent of Group 1 and 48% of Group 2 could read J2 at 25 cm unaided (P = .03). Sixty-four percent of Group 1 and 88% of Group 2 stated they could manage daily activities without glasses. However, 88% of Group 1 and 52% of Group 2 requested reading glasses to improve clarity of the smallest print. PMID- 8426317 TI - Keratometric comparison of 4.0 mm and 5.5 mm scleral tunnel cataract incisions. AB - Postoperative keratometric astigmatism was analyzed in 276 consecutive cases of phacoemulsification and posterior chamber lens implantation performed by one surgeon. The series was divided into two groups: one received 4.0 mm incisions and the other 5.5 mm incisions. Incision design was triplanar and required suture closure, i.e., all wounds were closed with two X-pattern 10-0 nylon sutures. Using the Cravy method, an insignificant difference in induced astigmatism was found between the groups at one day, two weeks, and one year after surgery. The average induced diopters of astigmatism for the 4.0 mm group was +0.80 +/- 0.94 at one day, +0.49 +/- 0.73 at two weeks, and -0.34 +/- 0.91 at one year; for the 5.5 mm group it was +0.69 +/- 1.07 at one day, +0.41 +/- 0.85 at two weeks, and 0.23 +/- 1.01 at one year. Two-week uncorrected visual acuity of 20/40 or better was similar for both groups. Seventy-six percent of the 4.0 mm group and 68% of the 5.5 mm group had acuities of 20/40 or better. A significant difference in the incidence of immediate postoperative hyphema was noted. The overall hyphema incidence was 9%, with a 14% and 4% incidence for 5.5 mm and 4.0 mm incisions, respectively. PMID- 8426318 TI - Pseudoaccommodation with the 3M diffractive multifocal intraocular lens: a refraction study of 52 subjects. AB - A refraction study was performed of 52 pseudophakic subjects implanted with a 3M diffractive intraocular lens (IOL). Twenty patients implanted with conventional monofocal IOLs from 3M were used as controls. The refraction for distance and near was determined, corresponding to the refractive (distance vision) and diffractive (near vision) foci. Pseudoaccommodation amplitude was measured relative to the variation of visual acuity as a function of "fogging" obtained with plus and minus lenses over any required spherical cylindrical power. Results showed that the 3M diffractive IOL acted as a bifocal lens in all subjects except those whose IOLs had decentered markedly. The dioptric range allowing good vision for distance and near is much wider with the diffractive IOL than it is with a monofocal IOL. Almost perfect vision, and the satisfaction of the patients, was achieved when the distance refraction was near to emmetropia and astigmatism was minimal. However, hyperopic biometric inaccuracies were less disturbing than myopic inaccuracies. PMID- 8426319 TI - Using the surgical result in the first eye to calculate intraocular lens power for the second eye. AB - This paper examines the possibility of using the surgical result in the first eye when planning the intraocular lens power for the second eye. Two methods were considered: (1) an empirical method by which one regards the second procedure as a repeat of the one in the first eye and calculates the power from the actual refractive error obtained in the first eye and (2) a theoretical method by which one measures the pseudophakic anterior chamber depth (ACD) of the first eye and uses this value to plan for the second eye. Based on the data from 136 second eye procedures using extracapsular cataract extraction, the prediction error of the empirical method ranged from -10.5 to +9.5 diopters. The error of the theoretical method ranged from -2.3 to +2.8 diopters, which was significantly more accurate than the empirical method (P < .001). We conclude that the fellow eye ACD may be used as a guideline for the assumed ACD of the second eye. However, such use of the fellow eye ACD could not be shown to improve power calculation predictions significantly. PMID- 8426320 TI - Calculation of the thickness of an intraocular lens. AB - Lens manufacturers do not usually supply information about the central thickness of an intraocular lens optic. This paper describes a method to calculate the central lens thickness from variables normally supplied by the manufacturer, i.e., the total lens power in situ, the edge thickness, the optic diameter, and some knowledge of the optic architecture. The formulas are universal and valid for any lens, regardless of lens design and material. There is a correlation between calculated values of optic thickness and values supplied by manufacturers. Individually estimated lens thickness may increase the accuracy of intraocular lens power calculation, pseudophakic anterior chamber depth estimation, and pseudophakic axial length calculation. PMID- 8426321 TI - Topographic analysis of early changes in corneal astigmatism after cataract surgery. AB - Using the Topographic Modeling System (TMS), we studied the early changes in anterior corneal curvature following cataract surgery. Thirty patients who had planned extracapsular cataract extraction (p-ECCE) or phacoemulsification were evaluated with TMS at three days, and one, two, three, and five weeks postoperatively. After p-ECCE, the upper part of the cornea which corresponded to the wound and the lower cornea markedly steepened and showed a bow-tie pattern of astigmatism. Eight (57%) of 14 cases showed little reduction in the corneal steepening at five weeks postoperatively. In five cases (36%) the steepening in the lower cornea was less than the minimal change in the upper cornea. After phacoemulsification, corneal steepening was much less than it was after p-ECCE; three cases (17%) showed almost no surgically induced alteration of the cornea. In eight phacoemulsification cases (61%), postoperative corneal steepening decreased rapidly and the corneal shape recovered within five weeks. Our topographic analysis clearly demonstrated that a smaller wound in phacoemulsification surgery produced less corneal steepening and stabilized more rapidly than a larger wound in p-ECCE. PMID- 8426322 TI - Comparative study of a collagen corneal shield and a subconjunctival injection at the end of cataract surgery. AB - To compare the effectiveness of subconjunctival injections and collagen shields in delivering anti-inflammatory agents and antibiotics after cataract surgery, we conducted a prospective study of 61 patients. They were randomly assigned to three groups: subconjunctival injection of gentamicin and dexamethasone; the same route plus a collagen shield without drug; collagen shield soaked in gentamicin and dexamethasone. All subjects had a manual extracapsular cataract extraction with posterior chamber intraocular lens implantation. Eyes were evaluated at the end of surgery and 24 hours later. The collagen shield achieved a progressive effect on pain and a decrease in conjunctival redness after 24 hours. The occurrence of folds in Descemet's membrane was less frequent and aqueous flare less severe than when subconjunctival injections were used. No adverse effect was reported. We conclude that the collagen shield could be a safe, better, noninvasive technique because of its double action of bandage and enhancement of drug penetration. PMID- 8426323 TI - Reproducibility and validity of laser flare/cell meter measurements of intraocular inflammation. AB - Preoperative and postoperative anterior chamber reactions in a series of cataract surgery patients were measured with a Kowa FC-1000 laser flare/cell meter by two different technicians, and clinical assessments of inflammation were recorded. The average cell and flare readings by the two technicians were nearly identical at every time point, showing the laser flare/cell measurements to be highly reproducible. The correlations between laser flare/cell measurements and clinical assessments at postoperative time points were highly positive (P < .01), demonstrating the validity of the laser flare/cell measurements. PMID- 8426324 TI - Removal of lens epithelial cells following loosening of the junctional complex. AB - We previously reported a new method to remove residual lens epithelial cells- dispersion aspiration. The cells were loosened from their junctional complexes with Dispase, a proteolytic enzyme. To avoid intraocular tissue damage, the enzyme preparation was dissolved in sodium hyaluronate and injected into the capsular bag, which was carefully preserved during endocapsular cataract surgery. The cells were then removed by minimum irrigation/aspiration. In this study we incorporated ethylenediaminetetraacetic acid (EDTA), a calcium chelating agent for separating epithelial cells in tissue culture, into the procedure. The results of experiments in vitro and in rabbits suggest that this procedure also removed cells effectively with negligible damage to the zonules and corneal endothelium. PMID- 8426325 TI - Results and complications of secondary intraocular lens implantation. AB - The results and the postoperative complications of 107 secondary intraocular lens implantations were reviewed over a two-year period. In 84.1% of the cases, postoperative visual acuity was the same or better than preoperative visual acuity with aphakic glasses or contract lenses. The most frequent postoperative complications were secondary glaucoma, six cases (5.6%); cystoid macular edema, five cases (4.6%); and bullous keratopathy, three cases (2.8%). PMID- 8426326 TI - Indications for and results of a large series of intraocular lens exchanges. AB - We conducted a retrospective review of 79 patients who had intraocular lens (IOL) explantation and exchange over a 12-year period. Seventy-nine eyes of 40 males and 39 females from 17 to 94 years of age were followed from three months to more than 12 years. Sixty-one percent (61%) were posterior chamber lenses and 39% were anterior chamber lenses replaced by 76% posterior chamber and 24% anterior chamber lenses. The indications for lens exchange were eccentric or displaced IOL (41.7%), endothelial decompensation (27.7%), incorrect IOL power (12.6%), and uveitis-glaucoma-hyphema syndrome (10.0%). Analysis of the clinical results revealed that 72% of the cases had postoperative visual acuity better than or equal to 20/30, and 8% had a loss of one or more lines of visual acuity. Among the complications occurring after IOL exchange were retinal detachment, glaucoma, corneal decompensation, and anisometropia. PMID- 8426327 TI - Peribulbar anesthesia. A prospective statistical analysis of the efficacy and predictability of bupivacaine and a lignocaine/bupivacaine mixture. AB - The object of this trial was to determine the efficacy in peribulbar anesthesia of bupivacaine 0.5%, 0.75%, and a combination of bupivacaine 0.5% and lignocaine 2% in equal parts. Fifty-one and 50 patients in random order were injected with 0.5% and 0.75% bupivacaine, respectively, and another 50 patients with the lignocaine/bupivacaine mixture. The results were recorded and all data were statistically analyzed. We concluded that the peribulbar route with the agents used in this trial was not a successful technique. Our criterion for a successful block, i.e., akinesia, was not achieved in more than 54% of cases. PMID- 8426328 TI - Progressive constriction of the anterior capsular opening following intact capsulorhexis. AB - The continuous curvilinear capsulorhexis has rapidly increased in popularity as the procedure of choice when using phacoemulsification for cataract extraction. Only recently, however, have complications of this technique been reported. We review the complications of a continuous curvilinear capsulorhexis and present three cases involving progressive constriction of the postoperative anterior capsular opening. One patient had a history of myotonic muscular dystrophy, another had pars planitis, and the third had high myopia. A review of ocular findings in myotonic dystrophy and pars planitis is also presented, and the possible pathophysiology of this progressive constriction is explored. PMID- 8426329 TI - Histopathology of acute intraoperative suprachoroidal hemorrhage associated with transscleral intraocular lens fixation. AB - An 83-year-old hypertensive female had penetrating keratoplasty with insertion of a transsclerally fixated posterior chamber intraocular lens for pseudophakic corneal edema associated with a closed-loop, semiflexible anterior chamber intraocular lens. An acute intraoperative suprachoroidal hemorrhage developed five minutes after passage of the transscleral sutures. The eye became blind and painful and was enucleated three weeks later. Histopathologic examination showed an idiopathic suprachoroidal hemorrhage with no evidence of disruption of the ciliary body vasculature. Acute intraocular hemorrhage resulting from a needle pass through the highly vascularized ciliary body has been a greatly feared, though as yet unreported, complication of this procedure. Damage to the anterior uveal circulation was not implicated as a cause of the hemorrhage in this case. PMID- 8426330 TI - Bilateral high hyperopia after radial keratotomy. AB - A patient with myopia and astigmatism in the ideal range for correction by radial keratotomy had surgery on both eyes. Postoperatively, she developed bilateral high hyperopia, with a +9.00 diopter refractive error (OD) and a spherical equivalent of +8.75 diopters (OS). Her best corrected visual acuity dropped one line and hyperopia increased during the follow-up. PMID- 8426331 TI - Assessment of the blood-aqueous barrier by fluorophotometry following poly(methyl methacrylate), silicone, and hydrogel lens implantation in rabbit eyes. AB - Intraocular lenses of silicone, hydrogel, or acrylic materials that can be implanted through small incisions are being proposed as an alternative to conventional poly(methyl methacrylate) lenses. The potential of each of these materials to stimulate intraocular inflammation is important in their selection as an implantable material. To investigate the potential of each material, we assessed by clinical slitlamp examination, fluorophotometry, and histopathology the inflammatory response induced in the rabbit eye following phacoemulsification and implantation of hydrogel, silicone, or poly(methyl methacrylate) intraocular lenses. All lenses seemed to be equally well tolerated. In general, the degree of inflammation seen clinically decreased over the four-month study; however, anterior segment fluorophotometry showed continued mild interruption of the blood aqueous barrier in all lens groups. Fluorophotometry is a sensitive method to assess persistent subclinical anterior segment inflammation. PMID- 8426332 TI - Posterior subcapsular cataracts associated with megestrol acetate therapy. AB - Megestrol acetate, used for eight weeks as an appetite stimulant in a man infected with human immunodeficiency virus, was associated with the development of bilateral posterior subcapsular cataracts. We propose that the glucocorticoid properties of megestrol were contributory. Complicating this association, the patient developed cytomegalovirus retinitis approximately six weeks after the cataracts were recognized. PMID- 8426333 TI - Small incision trabeculotomy combined with phacoemulsification and intraocular lens implantation. AB - Cataract surgeons frequently face the challenge of treating an open-angle glaucoma patient presenting for cataract surgery and intraocular lens implantation. It seems logical to treat both problems during the same surgical procedure if it can be done without jeopardizing the results of either. We present a short historic overview of the trabeculotomy procedure in adults, together with our technique using the new Fukasaku modification of the instrumentation (probes and forceps) and report 50 consecutive cases with a 100% three-month follow-up. The mean preoperative intraocular pressure was 19.3 mm Hg (range 16.0 to 37.0 mm Hg); three months after surgery it was 14.2 mm Hg (range 6.0 to 24.0 mm Hg). The mean preoperative pressure-reducing medications were 2.3 drugs per patient, and the postoperative mean was 0.7. Except for three cases of small postoperative hyphemas, no other complications were encountered during the study. PMID- 8426334 TI - Combined small incision phacoemulsification and trabeculectomy. AB - A method of combined cataract extraction and trabeculectomy is described in which phacoemulsification is performed through a 3.5 mm incision with insertion of a foldable hydrogel lens. This procedure is suitable for most cases in which glaucoma and cataract are coincident. In an initial series of ten patients followed for nine months, all those without additional pathology achieved a corrected visual acuity of 20/40 or better. The maximum induced astigmatism was 0.5 diopters (D), with no change or a reduction in astigmatism in seven patients (70%). Astigmatic shift between week 1 and three months was minimal (average = 0.075 D), with little change thereafter. Intraocular pressures were controlled (< or = 21 mm Hg) without adjunctive medication in all but one of the patients at nine months. This combined procedure preserves the principal advantages of small incision phacoemulsification: low induced astigmatism, early refractive stabilization, and rapid visual rehabilitation. In addition, reducing the extent of tissue dissection may reduce the stimulus to wound healing and early filtration failure. PMID- 8426335 TI - Oxygen desaturation during endoscopy in the elderly. AB - Arterial oxygen desaturation during oesophago-gastro duodenoscopy (OGD) is well recognised. It has been suggested that severe desaturation (greater than 7%) may predispose patients with cardiopulmonary disease and the elderly to cardiac arrhythmias. During OGD, of 106 elderly patients 26 developed ventricular and/or supraventricular ectopics, but these were not related to the degree of oxygen desaturation induced in this study. Apart from one episode of vasovagal syncope, which responded to intravenous atropine, no serious arrhythmias were recorded. Arterial oxygen desaturation during OGD was easily preventable with oxygen administration via nasal cannulae and was not associated with any adverse haemodynamic effects. Continuous cardiac and oxygen saturation monitoring should be routine practice in order to identify such problems. PMID- 8426336 TI - Delays to thrombolysis in the treatment of myocardial infarction. AB - In-hospital delays to thrombolysis were significantly shorter when thrombolysis was available on admission to the accident and emergency department than after transfer to the coronary care unit (median 60 min v 84 min, p < 0.0001). With direct admission by general practitioners to a coronary care unit the subsequent in-hospital delay was shorter (median 39 min p = 0.0004), but overall delay to thrombolysis longer (median 220 v 170 min, p = 0.0019) because of longer pre hospital delays. Overall delay was shortest with emergency ambulance referral and thrombolysis being administered in the accident and emergency department. PMID- 8426337 TI - Dupuytren's contracture in pensioners at the Royal Hospital Chelsea. AB - Dupuytren's disease of the hands was present in 55 (13.75%) of the 400 elderly ex servicemen living at the Royal Hospital Chelsea. Five men (9.1%) reported the condition in either a parent or sibling, but none was aware of an affected child. The prevalence of heavy drinking, non-insulin dependent diabetes or manual occupation was statistically the same in those with or without the condition. Overall, both hands were equally affected, but they differed in severity in 29 men. In milder cases the left hand was the more severely affected (grades 1 and 2); the reverse was true when the difference in severity was greater (grade 3). Since the original description of Dupuytren's disease [1] attempts have been made to link it with a variety of other conditions. These have included trauma, alcoholism and cirrhosis, [2] epilepsy [3] and diabetes mellitus [4]. A genetic link is accepted [5]. The condition is more common in men than women and becomes increasingly common with advancing age [6]. The prevalence in elderly men has been reported as high as 28.9%, rising to 64.3% in surveys of affected families [7]. There are still many physicians who, with a knowing wink when they spot that one of their patients has a Dupuytren's contracture, mentally register that he is an alcoholic. This paper is an attempt to disprove this fairy story. It describes a survey of Dupuytren's disease in a population of elderly men drawn from all parts of the United Kingdom and Ireland. PMID- 8426338 TI - Automated external defibrillators: defining optimum levels of accuracy based on the clinical practice of consultant cardiologists. AB - Several consultant cardiologists were invited to assess the accuracy of the Lifepak 200 automated external defibrillator (AED) (Fig. 1), from ECG records collected from pre-hospital cardiac arrest victims. They were asked to classify the ECG rhythms, and also give an opinion on whether or not a shock should have been given, and the potential harm of inappropriate treatment. As there was no absolute agreement between cardiologists in rhythm classification, sensitivity of the AED for ventricular fibrillation varied from 78% to 100%, and the specificity was between 92% and 100% according to each cardiologist. They agreed that all ventricular fibrillation should be shocked and failure to do so would reduce a patient's chances of survival; and that all other rhythms, and asystole, should not be shocked. Most experts believed shocking asystole would not be harmful, but opinions regarding the potential harm of administering shocks to patients with pulseless rhythm were mixed. PMID- 8426339 TI - A clinical audit of cardiopulmonary resuscitation. AB - The structure, process and outcome of cardiopulmonary resuscitation (CPR) at one hospital have been reviewed to determine where failings in the system could be improved, whether the existing training programme was adequate and well directed, and whether survival rates were reasonable. In the first seven months 143 reports were received; 49% of them were cardiac, 17% respiratory, and 24% mixed cardiorespiratory arrests. Overall, immediate survival was 43% of those with cardiorespiratory arrests. Seventy per cent of the immediate survivors were alive at 24 hours, and 69% of them survived to six weeks or beyond. The most important indicator for a successful outcome was the heart rhythm at the time of the resuscitation team's arrival, with 71% surviving from ventricular fibrillation, 20% surviving from apparent asystole, and only 9% surviving from electromechanical dissociation. No patient with the latter two rhythms survived to six weeks or left hospital. Neither the location or time of the arrest, nor the patient's age influenced the immediate survival. The audit revealed a need to improve telephone connections to the hospital switchboard, modify some equipment, and improve knowledge of the geography of the hospital site. Changes in emphasis in the training programme will alter the process of resuscitation. Outcome measures indicate that survival figures are comparable with published data, including recent data which act as a benchmark for quality control. PMID- 8426340 TI - Thrombolysis in acute myocardial infarction: an audit of practice. AB - A retrospective casenote audit of the use of thrombolysis in 212 of 335 patients with acute myocardial infarction (63%) was carried out on the coronary care unit of a Sheffield teaching hospital. The overall inpatient mortality was 17%, 12% in the thrombolysis group and 26% in the remainder (p < 0.005). Patients aged 75 years or older were less likely to die if they received thrombolytics (p < 0.05) despite an excess of anterior myocardial infarctions in the thrombolysis group (p < 0.01). The infusion of streptokinase had to be stopped in five patients (4%) because of allergy or arrhythmias. Thrombolytic therapy was considered in all patients with acute myocardial infarction; it was withheld inappropriately in eight cases and given inappropriately in two. Thrombolysis produced clear benefits in mortality for all patients, including those over 75 years old. There was a high rate of use of thrombolysis which was generally safe and appropriate, although adjustments to the guidelines may allow even more patients to receive this therapy. PMID- 8426341 TI - The ethics of itemised payment for clinical research procedures. PMID- 8426342 TI - Appraisal and junior medical staff. AB - A group of junior medical staff in a non-teaching district were surveyed by questionnaire to see how many had been appraised during their postgraduate training and to ascertain their attitude to appraisal. Of the 76% who replied 23% had had a formal appraisal but only 5% had had a structured interview; a further 42% had had an informal chat but 34% had never had any meaningful feedback. Nearly all the responders wished to be appraised on a wide range of skills, 66% wanting appraisal both during and at the end of the post held. The value of appraisal in stress management, improving performance and its relation to audit are discussed. PMID- 8426343 TI - Cardiopulmonary resuscitation: effect of training junior house officers on outcome of cardiac arrest. AB - The cardiopulmonary skills of one intake of junior house officers (JHOs) were assessed and all then underwent a six hour training course. In the following six months we studied the outcomes of 83 cardiopulmonary resuscitation attempts and compared them with results obtained in our hospital two years earlier as part of a multicentre survey when no formal training was undertaken. The JHOs performed poorly in all assessments before training. The initial and long-term survival rates in the present study were no better than in the earlier. There were 12 long term survivors, 11 required defibrillation. Decreasing the time taken to defibrillation and training other members of the arrest team in addition may lead to improved survival. The training of junior house officers alone is not an effective strategy for improving survival from cardiac arrest. PMID- 8426345 TI - Black looks ... at paternalism. PMID- 8426344 TI - Growth in prescription medicines expenditure: a European comparison. PMID- 8426346 TI - Interhealth: the WHO integrated programme for community health in non communicable diseases. PMID- 8426347 TI - Can we afford medical advances? AB - Medical advances, particularly the very expensive new therapeutic agents, will almost certainly not be affordable unless additional mechanisms are developed to deal with their introduction and subsequent use within the NHS. The granting of a product licence should be based on cost-effectiveness as well as on clinical efficacy and safety as proven by controlled clinical trials. The freedom of individual practitioners to prescribe both in general practice and in hospitals has to be restricted as far as the very expensive drugs are concerned. PMID- 8426348 TI - What is it like to have an angioplasty? PMID- 8426349 TI - Fund raising for the uninitiated. PMID- 8426350 TI - From Inkerman Street to Australia. The MEDICINE-Gilliland Fellowship. PMID- 8426351 TI - Artificial nutritional support in clinical practice in Britain. AB - Malnutrition is common in hospitalised patients. It often develops insidiously and its diagnosis is frequently delayed or missed. A multidisciplinary nutrition support team can improve the quality of nutritional support, reduce inappropriate feeding, reduce the complications associated with enteral and parenteral nutrition, and so improve clinical outcome and reduce hospitalisation. These improvements have obvious financial advantages, yet only a minority of British hospitals (25-30%) has a nutrition team. The increasing use of parenteral and enteral nutrition at home, which represents one of the most important areas of recent developments in artificial nutritional support, also has financial and clinical advantages, but the management of such patients is also less than optimal. Better education and greater awareness of nutritionally related problems, as well as changes in the local and national infrastructure of nutrition support services, are required to improve the quality of care and the clinical outcome for patients being treated by parenteral and enteral nutrition in hospital and at home. PMID- 8426352 TI - George Carmichael Low FRCP: an underrated figure in British tropical medicine. PMID- 8426353 TI - The visitors' book in the Khartoum Medical College. PMID- 8426354 TI - Heart failure in a DGH. PMID- 8426355 TI - In pursuit of a dubious principle. PMID- 8426356 TI - In pursuit of a dubious principle. PMID- 8426357 TI - The MD thesis in the training of a consultant physician. PMID- 8426359 TI - Who will care for our elderly people? PMID- 8426358 TI - Generic substitution. PMID- 8426361 TI - Highly selective tripeptide thrombin inhibitors. AB - Tripeptide aldehydes such as Boc-D-Phe-Pro-Arg-H (51) exhibit potent direct inhibition of thrombin. This distinction offers important insight for the design of more potent and selective serine protease inhibitors which may be useful pharmacological tools and hold promise for development of clinically useful agents. The structure-activity relationships (SAR) on a series of anticoagulant peptides with high selectivity for the enzyme thrombin are discussed. The SAR is centered on a series of di- and tripeptide arginine aldehydes based on the structure of 51. The structural and conformational role of the amino acid residue in position 1 was investigated by substitution with conformationally restricted aromatic amino acids, aromatic acids, and a dipeptide isostere containing the psi[CH2N] amide bond replacement. Many of these peptides demonstrate potent antithrombotic activity along with selectivity toward thrombin, determined by comparison of in vitro inhibitory effects on trypsin, plasmin, factor Xa, and tissue plasminogen activator. Compound 5f, D-1-Tiq-Pro-Arg-H.sulfate is highly active and the most selective tripeptide aldehyde inhibitor of thrombin reported to date. PMID- 8426360 TI - Enantiospecific syntheses of alpha-(fluoromethyl)tryptophan analogues: interactions with tryptophan hydroxylase and aromatic L-amino acid decarboxylase. AB - alpha-Fluoromethyl amino acids are enzyme-activated irreversible inhibitors of amino acid decarboxylases. Aromatic L-amino acid decarboxylase (AADC) is the enzyme responsible for the final step in the biosynthesis of both dopamine and serotonin via decarboxylation of L-dopa and 5-hydroxy-L-tryptophan, respectively. Our goal is to utilize antagonists of the serotonin-producing enzymes (tryptophan hydroxylase and AADC) as the basis for a chemotherapeutic approach to the treatment of carcinoid tumors, a rare tumor type characterized by the overproduction of serotonin. We report here an enantiospecific synthesis of alpha(S)-(fluoromethyl)tryptophan [(S)-11a] and alpha(S)-(fluoromethyl)-5 hydroxytryptophan [(S)-11b], as well as the (R)-enantiomers, based upon recent methodology involving the face-selective alkylation of cyclic tryptophan tautomers. Our synthetic route provided both enantiomers of 11a and 11b with greater than 97% enantiomeric purity based upon evaluation of the NMR spectra of their Mosher's acid derivatives. (S)-11a was evaluated as a substrate for P815 tryptophan hydroxylase and determined to have an apparent Km of 4.31 +/- 1.07 mM, essentially half the value previously reported for the racemic mixture of 11a with rat brain stem tryptophan hydroxylase. (R)-11a was not a substrate for P815 tryptophan hydroxylase. (S)-11b was evaluated as an enzyme-activated irreversible inhibitor of murine liver AADC and determined to have a KI of 24.3 +/- 3.01 microM and a k2 of 2.26 +/- 0.44 min-1. (R)-11b was not an inhibitor of murine liver AADC. PMID- 8426362 TI - Symmetry-based inhibitors of HIV protease. Structure-activity studies of acylated 2,4-diamino-1,5-diphenyl-3-hydroxypentane and 2,5-diamino-1,6-diphenylhexane-3,4 diol. AB - The structure-activity relationships in two series of novel, symmetry-based inhibitors of HIV protease, the enzyme responsible for maturation of the human immunodeficiency virus, are described. Beginning with lead compounds 3-6, the effect of adding polar, heterocyclic end groups to one or both ends of the symmetric or pseudosymmetric inhibitors was probed. Aqueous solubility was enhanced > 1000-fold while maintaining potent inhibition of purified HIV-1 protease and anti-HIV activity in vitro. Pharmacokinetic studies in rats indicated a substantial difference in the absorption properties of mono-ol-based and diol-based inhibitors. The oral bioavailability of inhibitor 19 in rats was 19%; however, the Cmax obtained failed to exceed the anti-HIV EC50 in vitro. Substantial plasma levels of potent inhibitors of the diol class were not obtained after oral administration in rats; however, the optimal combination of aqueous solubility and in vitro antiviral activity of several inhibitors support their potential use in intravenous therapy. PMID- 8426363 TI - Synthesis and in vitro evaluation of 5,6,7,8,9,10-hexahydro-7,10 iminocyclohept[b]indoles: high-affinity ligands for the N,N'-di-o-tolylguanidine labeled sigma binding site. AB - A series of 5,6,7,8,9,10-hexahydro-7,10-iminocyclo[b]indoles substituted at the 5 and/or 11 positions was synthesized from tropinone. Affinity for sigma binding sites was determined using [3H]-N,N'-di-o-tolylguanidine ([3H]DTG) and [3H]-(+)-3 (3-hydroxyphenyl)-N-1-propylpiperidine ([3H]-(+)-3-PPP) and for the dopamine D2 receptor labeled with [3H]sulpiride. Nearly all compounds studied in this series possessed a higher affinity for [3H]DTG than [3H]-(+)-PPP-labeled sigma sites, suggesting that [3H]DTG and [3H]-(+)-3-PPP radioligands label pharmacologically distinct sigma binding sites, as reported previously. Substitution at the 11 position with side chains containing a four-carbon tether resulted in compounds having the highest affinity for the [3H]DTG-labeled sigma site. The most potent and selective member of this series was 11-[4-(2-furanyl)butyl]-5,6,7,8,9,10 hexahydro-7,10-iminocyclohept [b] indole (40). Enantioselectivity was investigated by preparing the (+)- and (-)-isomers of 40. These studies revealed that (+)-40 was more potent at the [3H]-DTG-labeled sigma site whereas (-)-40 had a higher affinity at sigma sites labeled with [3H]-(+)-PPP. Racemic 40 was observed to possess a higher affinity than either of its respective enantiomers at both the [3H]DTG- and [3H]-(+)-3-PPP-labeled sites, suggesting an allosteric interaction. PMID- 8426364 TI - Inhibition of the tetracycline efflux antiport protein by 13-thio-substituted 5 hydroxy-6-deoxytetracyclines. AB - A series of 13-(alkylthio) and 13-(arylthio) derivatives of 5-hydroxy-6 deoxytetracycline (tetracycline, Tc) were synthesized and compared to the clinically used antibiotics tetracycline, methacycline, and minocycline for their ability to inhibit tetracycline efflux in an everted membrane vesicle assay of bacterial resistance to tetracyclines. The assay screened for the ability of tetracycline analogues to inhibit [3H]tetracycline uptake into everted membrane vesicles, thereby evaluating the molecular prerequisites for inhibition of an efflux-dependent resistant bacterial system. Thiol adducts attached at the exocyclic C13 carbon of methacycline led to an increase in inhibitor potency as compared to the reference antibiotics. The most potent inhibitors of [3H]tetracycline uptake into everted vesicles among these analogues, particularly members of the alkyl series, revealed important structure-activity relationships between inhibitor potency, steric parameters, and lipophilicity at the C13 sulfur position. PMID- 8426365 TI - Novel 6-alkoxypurine 2',3'-dideoxynucleosides as inhibitors of the cytopathic effect of the human immunodeficiency virus. AB - Twenty-one 6-alkoxypurine 2',3'-dideoxynucleosides were enzymatically synthesized with nucleoside phosphorylases purified from E. coli. Eighteen analogs exhibited anti-HIV-1 activity in MT4 cells. Two analogs, 6-(hexyloxy)-(17) and 6 (heptyloxy)-(18) purine 2',3'-dideoxynucleoside, were as potent as 2',3' dideoxyinosine (ddI, didanosine, Videx). Although the antiviral activities of 17 and 18 were equivalent, 18 was more cytotoxic. Analogs containing less than four carbons in the 6-alkoxypurine substituent exhibited weak anti-HIV-1 activity. Analogs containing more than seven carbons in the 6-alkoxypurine substituent were too cytotoxic to be effectively evaluated for antiviral activity. Several 6 alkoxypurine 2',3'-dideoxynucleosides were evaluated for substrate activity with calf intestinal adenosine deaminase (ADA). Increasing the carbon chain length of the 6-alkoxypurine substituent decreased the rate of dealkoxylation. The best substrate in this series was 6-methoxypurine 2',3'-dideoxynucleaside (1); however, the rate of dealkoxylation of 100 microM 1 was 0.17% of the rate of deamination of 100 microM 2',3'-dideoxyadenosine. Compound 17, the most potent anti-HIV-1 analog, was not a substrate for ADA. EHNA (erthro-9-(2-hydroxy-3 nonyl)adenine), a potent inhibitor of ADA, had little effect on the antiviral activities of 17 and ddI. In contrast, coformycin, a potent inhibitor of both ADA and AMP deaminase, dramatically decreased the antiviral activity of 17, but not the antiviral activity of ddI. Thus, AMP deaminase appeared to be involved in the anabolism of 17. The pharmacokinetic profile of 17, the most promising analog in this series, was determined in the rat. At least seventeen metabolites of 17, including ddI, were detected in plasma samples. This analog also had poor oral bioavailability. PMID- 8426366 TI - Defining structural requirements for neuropeptide Y receptors using truncated and conformationally restricted analogues. AB - To further elucidate the minimum bioactive conformation of neuropeptide Y (NPY), a series of truncated and conformationally constrained analogues has been prepared. The synthesis and purification of these peptides was achieved using routine laboratory strategies and techniques. Parent molecules consisted of the native NPY N-terminal 1-4 and C-terminal 25-36 segments, having the residue 5-24 core replaced by either a single flexible omega-aminoalkanoic acid, or a more rigid Pro-Gly or Pro-DAla sequence which was expected to constrain a putative turn, and allow the N- and C-termini to align. Cross-linking between residues 2 and 27 through lactamization using side-chain length and chirality suggested by computer simulations, resulted in cyclo-(2/27)-des-AA7-24[Glu2,Gly6,DDpr27]NPY that exhibited very high affinity (Ki = 0.3 versus 0.3 nM for NPY) for the Y2 receptor using SK-N-BE2 human neuroblastoma cells, yet very low affinity for the Y1 receptor using SK-N-MC human neuroblastoma cells (Ki = 130 versus 2.0 nM for NPY). The added constraint resulting from bridging in this analogue as well as in others suggested that the combination of the deletion of residues 5-24 and the introduction of an internal ring produced exclusive selectivity for the Y2 receptor with little or no loss of affinity. The tolerance of structural recognition was further demonstrated as a second ring was introduced which was expected to constrain the amphiphilic alpha-helix, resulting in the full Y2 agonist dicyclo (2/27,28/32)-des-AA7-24 [Glu2,32,DAla6,DDpr27,Lys28]NPY. Improvement of Y1 binding activity was achieved only by including more residues (des-AA10-17) in the central PP-fold region, while allowing limited flexibility of the termini. Although the length of the bridge seemed to have little effect on binding potency, changes in the location of and chirality at the bridgehead resulted in analogues with different binding affinities. Combination of optimum structural modifications resulted in cyclo-(7/21)-des-AA10-18[Cys7,21]NPY, an analogue shortened by 25% but retaining comparable binding properties to that of native NPY at Y1 and Y2 receptor types (Ki = 5.1 and 1.3 nM, respectively). PMID- 8426367 TI - 32-Methyl-32-oxylanosterols: dual-action inhibitors of cholesterol biosynthesis. AB - Lanosterol 14 alpha-methyl demethylase (P-450DM) is the cytochrome P-450 monooxygenase which oxidatively removes the 14 alpha-methyl group of lanosterol. This demethylation is considered to be a rate-limiting step in the conversion of lanosterol to cholesterol. The intermediates in this transformation are known to bind very tightly to P-450DM and have been implicated in the regulation of 3 hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) activity, the rate-limiting enzyme in overall cholesterol biosynthesis. Three 32-methylated analogs of the intermediates generated during the removal of the 14 alpha-methyl group by P 450DM, compounds 17a, 17b, and 18, have been prepared and their biochemical activities assessed. All three compounds were found to be direct inhibitors of P 450DM. These compounds were also shown to suppress HMGR activity by reducing the level of enzyme protein. PMID- 8426368 TI - Rheumatoid arthritis, aspirin, and gastrointestinal cancer. PMID- 8426369 TI - Wing of fly, gene of worm: unusual models shed light on cancer. PMID- 8426370 TI - Australian study links certain analgesics to renal cancers. PMID- 8426371 TI - Rising breast cancer rates fueled by better detection. PMID- 8426372 TI - RAC approves policy for single-patient use of gene therapy. PMID- 8426373 TI - Potentiation of antitumor immunotoxins by liposomal monensin. AB - BACKGROUND: The cytotoxicity of specific ricin A-chain immunotoxins is greatly enhanced in vitro by the carboxylic ionophore monensin. However, the highly lipophilic nature of monensin, which is reflected in its poor solubility and short half-life, has restricted its use in in vivo animal studies. PURPOSE: The purpose of this study was to assess the ability of monensin incorporated in unilamellar vesicles (liposomes) to potentiate antitumor immunotoxins in vitro and in vivo. METHODS: Monensin was incorporated into liposomes and used in combination with specific immunotoxins against human tumor cell lines in vitro and in vivo. Inhibition of [3H]leucine incorporation was used to evaluate the cytotoxic action of immunotoxin with or without monensin in vitro on the following human tumor cell lines: H-MESO-1 malignant mesothelioma, LS174T colorectal carcinoma, and U373, U87, and MG-1 glioblastomas. For the in vivo studies of immunotoxins and liposomal monensin, BALB/c nu/nu mice were inoculated intraperitoneally with H-MESO-1 cells. RESULTS: Liposomal monensin potentiated the cytotoxic action of cell-specific anti-human transferrin receptor immunotoxin on H-MESO-1 target cells at a molar concentration of monensin that was 160-fold lower than the concentration of monensin in buffer that produced the same effect (0.3 nM versus 0.05 microM). Moreover, immunotoxin plus 0.1 microM liposomal monensin was fivefold more toxic for H-MESO-1 cells and 1000-fold and 2200-fold more toxic for human glioblastoma U373 and U87 cells, respectively, than immunotoxin plus 0.1 microM free monensin in buffer. Liposomal monensin produced similar effects when it was combined with different specific immunotoxins and other target cell lines (i.e., LS174T, U87, and CEM). Immunotoxin specificity was preserved with liposomal monensin, as shown by the absence of effect with non cell-binding immunotoxins or on antigen-negative cell lines. In mice, liposomal monensin in combination with specific immunotoxin substantially prolonged survival, and three (21%) of 14 mice bearing H-MESO-1 xenografts treated with the liposomes showed no evidence of tumor at day 160 after treatment. Treatment with control immunotoxin plus liposomal monensin was ineffective. CONCLUSION: These findings suggest that encapsulation of monensin into liposomes increased the capacity of monensin to enhance the potency of cell-specific immunotoxin in vitro and in vivo. PMID- 8426374 TI - Incidence of cancer among patients with rheumatoid arthritis. AB - BACKGROUND: To evaluate hypotheses about the relationship between immune alterations and cancer, several investigators have determined cancer incidence in groups of patients with rheumatoid arthritis (RA), a chronic autoimmune disease. The primary finding has been an increased risk of hematopoietic cancers. PURPOSE: In this study, we have attempted to refine estimates of the association between RA and subsequent development of specific cancers. METHODS: We investigated site specific cancer risk associated with RA in a population-based cohort study of 11683 Swedish men and women with a hospital (inpatient) diagnosis of RA. These case patients were identified from 1965 to 1983 and had follow-up through 1984 by computer linkage of the Swedish Hospital Inpatient Register to the National Swedish Cancer Registry (840 case patients with cancer) and the Swedish Registry of Causes of Death. Cancer risk was estimated by standardized incidence ratios (SIRs) for specific cancers. RESULTS: For men and women overall, there were decreased risks for cancers of the colon (SIR = 0.63; 95% confidence interval [CI] = 0.5-0.9), rectum (SIR = 0.72; 95% CI = 0.5-1.1), and stomach (SIR = 0.63; 95% CI = 0.5-0.9) and an increased risk for lymphomas (SIR = 1.98; 95% CI = 1.5 2.6). CONCLUSIONS: The reduced risk for colorectal cancer in patients with RA is consistent with previous studies of RA patients and with reports which state that use of nonsteroidal anti-inflammatory drugs may protect against the development of large bowel cancers. The excess of lymphomas also confirms a number of earlier investigations of RA patients. PMID- 8426375 TI - Ventricular arrhythmia and adrenal insufficiency after fluorouracil: new side effects of an old drug. PMID- 8426376 TI - DDT and pancreatic cancer. PMID- 8426377 TI - Medicine's future challenges. PMID- 8426378 TI - Educating the African-American community on organ donation. PMID- 8426379 TI - Morbidity of low-birthweight infants with intrauterine cocaine exposure. AB - The effects of intrauterine cocaine exposure on 158 low-birthweight infants (500 g to 2500 g) were compared with 536 low-birthweight infants not known to be exposed to cocaine who were admitted to our neonatal intensive care unit. Univariate analyses indicated that drug-exposed infants had lower birthweights when compared with control infants. Frequency of necrotizing enterocolitis differed between the drug-exposed infants and controls (11% versus 6%). Frequency of severe respiratory distress syndrome, severe intraventricular hemorrhage, and bronchopulmonary dysplasia did not differ between groups of similar birthweight. We conclude that the most evident risks of prenatal cocaine exposure are low birthweight and increased incidence of necrotizing enterocolitis. PMID- 8426380 TI - Postoperative adult respiratory distress syndrome: the Howard experience. AB - To describe the natural history of adult respiratory distress syndrome (ARDS), we compared the clinical course of two groups of postoperative critically ill patients who developed respiratory failure requiring prolonged endotracheal intubation (minimum: 4 days). The two groups with ARDS (Group 1) and without ARDS (Group 2) were compared for the following clinical parameters: PO2/FIO2 ratio, cardiac index, pulmonary capillary wedge pressure, systemic and pulmonary vascular resistance, pulmonary compliance, serial chest radiograph findings, presence or absence of sepsis, oxygen transport parameters, morbidity, and mortality. Twenty patients are described over a 9-month period ending in March 1990-10 patients in each group. Characteristic differences between the two groups in terms of oxygen consumption, pulmonary compliance, pulmonary vascular resistance, numbers of organ systems failing, and mortality were noted. Our experience supports the recommendations of early institution of positive and expiratory pressure and optimizing oxygen delivery parameters in the management of ARDS. PMID- 8426381 TI - Hospital-acquired morbidity on a neurology service. AB - Clinical services must monitor hospital-acquired morbidity, but what rates are expected specifically for neurology inpatients is not evident from published studies. We studied prospectively 1317 consecutive admissions to a neurology service in a university-affiliated city hospital from 1987 to 1990 and recorded all nosocomial infections, nosocomial pneumonia, and decubitus ulcers of stage III or IV. Over the 3-year period, 6.8% of patients had > or = 1 nosocomial infection (and almost half of these had a nosocomial bloodstream infection); 3.1% had > or = 1 case of nosocomial pneumonia; 1.2% developed severe decubitus ulcers, and 8.4% had one or more of the three complications. The incidence of nosocomial infection exceeds that expected from multihospital studies. How much of the excess is peculiar to neurology patients and how much can be attributed to factors in our community and at our hospital cannot be determined from this study. Furthermore, our statistics are not meant as norms, but as initial estimates for quality assurance. PMID- 8426382 TI - Preliminary survivorship report on combined intraoperative radiation and hyperthermia treatments for unresectable pancreatic adenocarcinoma. AB - Five patients with unresectable pancreatic adenocarcinoma were treated with simultaneous intraoperative radiotherapy and intraoperative hyperthermia. Postoperative survivorship averaged 15.8 months, which compares favorably to a previous study in which 19 patients receiving intraoperative radiotherapy without intraoperative hyperthermia survived an average of 6.05 months. Three of the five patients following the experimental protocol of intraoperative radiotherapy and intraoperative hyperthermia with additional external beam radiotherapy are still alive, which may result in average postoperative survivorship exceeding 15.8 months. PMID- 8426383 TI - Erect lateral pelvimetry and the outcome of labor in Lagos. AB - The erect lateral radiograph pelvimetry measurements was related to the outcome of labor in 173 patients. The most common indication for pelvimetry was one previous caesarean section in 90 (52%) patients. Sixty-nine (39.9%) pelvimetry measurements were performed because of suspected cephalopelvic disproportion, while 20 (11.6%) were done because of breech presentation. In patients with a cephalic presentation at delivery, the mean +/- standard deviation (SD) obstetric conjugate for the 87 women who delivered vaginally (group 1) was 11.42 +/- 1.5 cm, while that of the 45 women who had emergency caesarean section (group 2) was 10.77 +/- 1.5 cm. The mean +/- SD anteroposterior outlet diameter for groups 1 and 2 were 12.24 +/- 1.9 cm and 11.84 +/- 1.7 cm respectively. Both differences were found to be statistically significant (p < 0.001 and p < 0.05 respectively). The mean +/- SD birthweight of the babies in groups 1 and 2 were 3.34 +/- 0.79 kg and 3.42 +/- 0.86 kg respectively. There was no statistically significant difference between the two groups (p < 0.05). The critical obstetric conjugate for safe vaginal delivery was 9.88 cm while the corresponding value for the anteroposterior pelvic outlet diameter was 10.24 cm. PMID- 8426384 TI - A racial difference in erythrocyte sedimentation. AB - Erythrocyte sedimentation rate (ESR) was examined in blacks and whites aged 18 to 74 years in the first National Health and Nutrition Examination Survey (NHANES I). In each age and sex group, median ESR was 2 mm/hr to 13 mm/hr higher in blacks than in whites even after exclusion of persons with extreme hemoglobin values or history of conditions affecting ESR. Apparently healthy blacks were more likely than whites to have elevated ESR greater than 20 mm/hr in women and 9 mm/hr in men. These data suggest a racial difference in ESR that is independent of age, hemoglobin concentration, and certain chronic diseases. PMID- 8426385 TI - Prevalence of chronic obstructive pulmonary disease in blacks. AB - Chronic obstructive pulmonary disease (COPD) is a significant cause of morbidity and mortality in blacks. The prevalence of COPD among blacks was estimated from the spirometry data obtained from the first National Health and Nutrition Examination Survey (NHANES), 1971-1975. Of 873 subjects, 585 (67%) had acceptable spirometry trials. Chronic obstructive pulmonary disease was defined as a forced expiratory volume in one second (FEV1) less than 65% of the predicted value. The mean FEV1 percentage predicted was 96.7%. The overall prevalence of COPD was 5.4%; 3.7% for males and 6.7% for females. The prevalence was significantly higher with age for both males and females. The multiple logistic regression analyses showed that age and sex were associated with COPD but respiratory symptoms did not attain statistical significance. PMID- 8426386 TI - The fate of the other 98. PMID- 8426387 TI - Attitudes of medical students and family practice residents toward geriatric patients. AB - The federal government, as well as teaching institutions, are concerned about the current negative attitudes of doctors, medical students, and paramedical personnel toward the elderly. Increased life expectancy at birth and lowered birth rates are changing the demographics of America. As the number of elderly citizens increases, greater demands are being placed on medical educators to train physicians who can meet the "geriatric imperative." The Institute of Medicine has recommended that comprehensive humanistic medical education in geriatrics be integrated throughout the curricula of medical schools. Research is needed to see if change can be implemented in physician training to improve attitudes toward the elderly. Previous attempts to improve medical students' attitudes toward the elderly have met with mixed success. Control groups have seldom been used. It is important to determine whether the effects of medical education extends beyond the immediate boundaries of a training curriculum. This article reports the results of a study on negative attitudes toward the elderly among residents, medical students, and physician's assistant students in the family medicine department at the King/Drew Medical Center in Los Angeles, California. PMID- 8426389 TI - A handheld calculator program for determination of gene transfer between races. AB - Methods of statistical analysis have permeated all biomedical fields, but the advent of inexpensive handheld calculators has enabled those individuals unskilled in advanced mathematics to make competent analyses. Statistics cannot be avoided in genetics and their evaluations, really estimated, are essential to further knowledge and understanding. Older scientists had to rely on the use of paper and pencil to derive formulas and then apply them. The experience gained in this one small area of investigation has been presented in detail to help other investigators who do not possess a broad mathematical background. PMID- 8426388 TI - Atypical carcinoid of the lung with cervical and submandibular lymphadenopathy: a case report. AB - A 33-year-old female presented with a 2-month history of right cervical and submandibular lymphadenopathy. She had recent onset of a nonproductive cough but was otherwise asymptomatic. The usual symptoms of carcinoid syndrome such as cutaneous flushing, abdominal pain, and asthma were absent. PMID- 8426390 TI - Acute pancreatitis in a child with sickle cell anemia. AB - A case of pancreatitis that occurred as a complication of a vaso-occlusive crisis in a child with sickle cell anemia is reported. We encourage others to consider pancreatitis as a cause for abdominal pain in children with multisystem diseases, particularly those that may cause ischemic organ injury such as sickle cell anemia. PMID- 8426391 TI - Effects of the white medical community on black health. PMID- 8426392 TI - Female urinary tract fistulas. AB - Urinary tract fistulas in women are an uncommon complication of a variety of surgical procedures. However, such fistulas lead to significant patient and physician distress, and have important medicolegal implications. Successful repair of urinary tract fistulas requires careful preoperative evaluation and adherence to basic surgical principles. A variety of approaches and techniques may be used, and the choice of procedure is less important than achieving adequate resection of fibrosis with watertight, tension-free closure of well vascularized tissues in layers. Interposition grafts of omentum, muscle, peritoneum and labial fat may be used in recurrent, complicated or radiated fistulas, and add significantly to the rate of success. Overall, successful repair of urinary tract fistulas can be achieved in the majority of cases. PMID- 8426393 TI - Calcium and calcium magnesium carbonate specimens submitted as urinary tract stones. AB - Of 8,129 specimens submitted as urinary stones from 6,095 patients, 67 from 15 patients were predominantly calcium carbonate or calcium magnesium carbonate (dolomite) by infrared analysis. Detailed study of 1 man and 4 women who submitted 3 or more such specimens showed that all were of aragonite calcium carbonate crystal form in 2 women and all calcite in the man. All 3 patients had a long history of nephrolithiasis preceding submission of calcium carbonate stones. There was frequent and often painful spontaneous passage of many small stones. Medullary sponge kidney was reported in 2 patients. Specimens submitted by the other 2 women included dolomite and quartz artifacts. Of the other 10 patients 4 had calcite and 1 had aragonite (possibly true stones). Five patients had artifacts with dolomite in 3 and mixed specimens in 2. True calcium carbonate kidney stones and calcium carbonate artifacts may be difficult to distinguish, and dolomite and quartz artifacts may require x-ray diffraction for clear-cut diagnosis. PMID- 8426394 TI - The transplantation of difficult donor kidneys and recipients: helpful surgical techniques. AB - Due to the current shortage of organ donors a coalescence of surgical techniques has been applied in an attempt to use all compatible organs that may have been discarded due to anatomical variations, disease or damage, and to transplant them into higher surgical risk recipients. Selected cases have been chosen that illustrate some of these techniques, and the various types of recipients and donor kidneys. Cases 1 and 2 illustrate applications of the orthotopic technique of renal transplantation, as well as reconstructive measures that may be necessary for success. Renal transplantation in perhaps one of the more commonly encountered difficult recipients, the patient with severe atherosclerotic vascular disease, is addressed by cases 1 and 3. The use of unusual donor organs, such as horseshoe kidneys and pediatric donor en block kidneys, is particularly pertinent in the attempt to transplant all viable organs and is demonstrated in cases 4 and 5. These cases illustrate the application of various surgical techniques in which either a difficult recipient or a damaged and/or diseased donor kidney may have otherwise precluded renal transplantation. PMID- 8426395 TI - Vitamin B12 deficiency in patients with ileocolic neobladders. AB - The terminal ileum is widely used for reconstruction of the lower urinary tract. Since the terminal ileum exclusively absorbs vitamin B12, removal of ileum from the gastrointestinal trace for use in lower urinary tract reconstruction may predispose patients to vitamin B12 deficiency. In a prospective study serial vitamin B12 levels were obtained postoperatively in 24 patients who underwent cystectomy for malignancy and ileocolic neobladder urinary diversion with a mean followup of 25 months (range 6 to 53). Any patient who had a low vitamin B12 serum level (less than 200 pg./ml.) underwent a Schilling test to confirm malabsorptive vitamin B12 deficiency. Six patients (25%) had low serum vitamin B12 levels, of whom 3 (13%) had an abnormal Schilling test. No patient had megaloblastic anemia. One patient with malabsorptive vitamin B12 deficiency had neurological symptoms 53 months postoperatively. We conclude that vitamin B12 deficiency can occur following ileocolic neobladder reconstruction. Patients with true vitamin B12 deficiency should be identified and placed on lifelong parenteral vitamin B12. PMID- 8426396 TI - The Mainz pouch II (sigma rectum pouch). AB - A low pressure rectosigmoid reservoir for urine is created obviating the need for colostomy, augmentation or extensive bowel surgery. Antimesenteric splitting of the intestine at the rectosigmoid junction and subsequent side-to-side anastomosis are performed. Urodynamic data demonstrate that the detubularization is effective in rendering high pressure bowel contractions ineffective. Without the risk of damaging the mesentery the pouch is fixed at the promontory, which lessens the risk of ureteral kinking and upper urinary tract dilatation. The technique is indicated not only in cases of failed ureterosigmoidostomy but also for primary urinary diversion. All 47 patients who underwent the operation were evaluable with a followup of 1 to 20 months (mean 10 months). All patients are continent during the daytime with a mean emptying frequency of 5 times. All but 1 elderly woman are dry at night with a mean frequency of 1 episode. With the reservoir full the basal pressure was 24 cm. water and the highest peak pressure recorded was 35 cm. water. The low pressure improves continence, protects the upper urinary tract and even allows dilated ureters to be implanted. PMID- 8426397 TI - How safe is 1% alum irrigation in controlling intractable vesical hemorrhage? AB - A prospective study was done to evaluate the efficacy and safety of intravesical instillation of 1% alum solution in 12 cases of hematuria of vesical origin, uncontrolled by saline irrigation for 24 hours via a 3-way Foley catheter. There were 10 cases of transitional cell carcinoma and 2 of radiation cystitis. Complete response was noted in 6 patients and a partial response in 4. Local side effects included suprapubic pain and vesical tenesmus, which were controlled by antispasmodic and/or analgesic drugs. Transient low grade pyrexia (maximum up to 38.2C) was noted in 4 patients. Among the other various clinical and biochemical parameters, serum aluminum level and prothrombin time showed statistically highly significant changes. Serum aluminum increased from an average baseline value of 1.68 to 3.36 mumol./l. without clinical evidence of aluminum toxicity and with levels well below the recommended safe limit. Prothrombin time increased parallel with the increase in serum aluminum level to a maximum of 1 1/2 times the control. Prothrombin values, therefore, can be used clinically, since they are readily obtainable whereas serum aluminum levels are not. Vesical irrigation with 1% alum solution is a safe method to control hematuria of vesical origin in properly selected cases. PMID- 8426398 TI - Fibronectin expression on surgical specimens correlated with the response to intravesical bacillus Calmette-Guerin therapy. AB - Attachment of bacillus Calmette-Guerin (BCG) organisms to the bladder during intravesical therapy is thought to be mediated exclusively by the glycoprotein fibronectin, which is expressed variably on epithelial surfaces and on basement membranes. We examined the relationship between the degree of fibronectin expressed on surgical specimens obtained from 50 candidates for BCG therapy and the subsequent clinical response. Immunoperoxidase staining for fibronectin was performed on tumor, nonadjacent normal mucosa and basement membrane tissues, and the intensity of the staining was scored on a scale of 0 to 3+ (control 2+). In the absence of recurrence at quarterly surveillance cystoscopy, a course of Tice BCG therapy consisted of 6 weekly and 12 monthly instillations. Recurrence of noninvasive tumor prompted a second BCG course. Followup ranged from 24 to 66 months (median 40 months). Of the 50 patients (11 with carcinoma in situ) disease progression occurred in 9 (none with carcinoma in situ). Compared to the results for tumors or for basement membranes, the degree of fibronectin expression on normal mucosa was well correlated with the clinical response (r = 0.59, p < 0.001 by Kendall Tau B). Routine assessment of fibronectin expression on the normal mucosa associated with superficial bladder cancer may be useful for predicting the clinical response to BCG therapy. PMID- 8426399 TI - The use of nuclear morphometry in predicting recurrence of transitional cell carcinoma. AB - Despite apparently adequate surgical resection, metastases develop in about 50% of the patients who undergo radical cystectomy for invasive bladder cancer and they die of disease within 2 years of surgery. Recent reports have shown that combination chemotherapy with regimens including cisplatin results in significant improvement in time to progression and overall survival for patients with transitional cell carcinoma. We have used quantitative nuclear morphometry in an effort to predict which individual patients with muscle invasive transitional cell carcinoma are at increased risk for recurrence and in need of adjuvant therapy. A total of 14 patients with deeply invasive tumors without known metastases (tumors, nodes and metastasis stage P3A-B, N-, M0) who had not received perioperative radiation or systemic chemotherapy was studied. Of the patients 7 (group 1) had no evidence of disease with a mean followup of 41 months and 7 (group 2) had recurrent transitional cell carcinoma at a mean followup of 17 months. Both groups were similar with respect to age and could not be distinguished by standard pathological analysis of stage (p = 0.66) or grade (p = 0.99). A total of 150 tumor nuclei from each patient was digitized and analyzed by a high resolution quantitative morphometric imaging system, developed in this laboratory, to determine if nuclear shape descriptors could separate these 2 clinically distinct groups. A multivariate analysis combining 3 independent nuclear shape descriptors (average nuclear area, minimum feret-diameter ratio and kurtosis of feret-diameter ratio) was highly significant (p = 0.003) and separated patients in group 1 from group 2 without overlap. These encouraging results suggest that nuclear morphometry may be valuable in determining which patients are at increased risk of disease recurrence following radical cystectomy and, therefore, who should receive adjuvant chemotherapy. PMID- 8426400 TI - The use of buccal mucosa patch graft in the management of anterior urethral strictures. AB - We describe our experience with 20 patients undergoing 1-stage correction of an anterior urethral stricture using a buccal mucosa patch graft. This technique was used for treatment of short strictures (1 to 2 cm.) that usually required a 2 to 4 cm. repair, making excision and end-to-end anastomosis impractical. Results were excellent in 18 patients, while 2 required revision for recurrent stricture. Urethrocutaneous fistulas and diverticulas were not encountered in our series. The buccal mucosa patch graft is hairless and, therefore, it can tolerate trauma and infection adequately. This technique represents a reasonable alternative when penile skin cannot be used or endoscopic manipulation is not indicated. PMID- 8426401 TI - Periurethral polytetrafluoroethylene paste injection in incontinent female subjects: surgical indications and improved surgical technique. AB - We present the results with 2 techniques for periurethral polytetrafluoroethylene (Polytef) injection in 21 female subjects with type III stress urinary incontinence. The standard technique included the use of a stainless steel needle for injection, paste "sopping" and a Wolff, Storz or Lewy syringe as an injecting element. Postoperatively, no catheters were left indwelling and all patients were encouraged to urinate following recovery from the anesthesia. The modified technique included the use of a 14F angio-catheter for injection of the paste, paste heating and a Lewy syringe or Mentor gun as injector. Postoperatively, all patients were left with an indwelling suprapubic catheter for 3 to 5 days. A total of 27 injections was performed, including 9 with the standard technique and the last consecutive 18 with the modified technique. Average followup has been 11.4 months. Cure, improvement and no change rates from the preoperative condition were 11%, 22% and 67% with the standard technique and 39%, 17% and 44% with the modified technique, respectively. In the latter group 3 patients had received pelvic radiotherapy as definitive treatment for pelvic malignancies. The overall failure rate in patients with a stable detrusor was 42% compared to 75% in the group with bladder instability and low compliance. Advantages of the modified technique include avoidance in the formation of intraoperative and postoperative fistulas, and easier handling and injection of the heated paste to achieve urethral compression. Improved short-term results with the modified technique indicate that a larger group of patients and long-term followup are essential requirements to determine the true efficacy of this technical modification. Based on these preliminary results, we now prefer the modified technique to the standard technique in the management of type III stress urinary incontinence. PMID- 8426402 TI - Use of a double cuff AMS800 urinary sphincter for severe stress incontinence. AB - Stress urinary incontinence in the male patient has been successfully treated with the artificial urinary sphincter. However, approximately 15% of the patients treated are still significantly wet despite some improvement with this device. These patients usually are almost totally incontinent before artificial sphincter implantation. Of 15 such patients 80% were rendered satisfactorily dry by adding a second sphincter cuff around the bulbous urethra. This double cuff technique increases the success rate with the AMS800 sphincter to greater than 95%. PMID- 8426403 TI - Dog bites to the male genitalia: characteristics, management and comparison with human bites. AB - Dog bites to the external male genitalia occur infrequently. We present 4 new cases and review 4 others described previously. Victims tend to seek medical care quickly. Thus, morbidity is directly related to the severity of the initial wound and delayed infectious complications appear to be minimal. Guidelines for management include irrigation, debridement as necessary, empiric antibiotics, consideration of tetanus and rabies immunization, and primary wound closure or surgical reconstruction. The differences between dog bites and human bites to the genitalia--primarily interval to presentation and subsequent likelihood of infection--are summarized. Measures to prevent dog bites are discussed. PMID- 8426404 TI - Long-term results with vacuum constriction device. AB - From November 1985 to April 1990, 216 consecutive patients were treated with the vacuum constriction device. Patients were mailed an initial questionnaire (group 1) and a long-term questionnaire (group 2) at a median followup of 3 and 29 months, respectively. Of 202 available patients 161 in group 1 (75%) and 115 in group 2 (57%) responded. Regular use of the vacuum constriction device was reported by 69% group 1 and 70% group 2 patients. patient and partner satisfaction was 82% and 87% in group 1, and 84% and 89% in group 2, respectively. There were no significant differences between the groups with respect to regular use and patient or partner satisfaction (p < 0.05). Quality of erection was evaluated for hardness, length and circumference, and with satisfaction greater than 90% in both groups. Median times per month of successful intercourse were 1, 4 and 4 for the year before, during and after obtaining the vacuum constriction device in group 2. Also, 79% of the patients in group 2 reported a statistically significant increase in the frequency of intercourse per month in the first year, which was sustained beyond the first year in 77% (p < 0.01). Our results support the efficacy of the vacuum constriction device for the treatment of impotence. Overall regular use rates as well as patient and partner satisfaction appear to be high. Furthermore, excellent initial results appear durable in most patients. PMID- 8426405 TI - Intraluminal device pressures in 3-piece inflatable penile prostheses: the "pathophysiology" of mechanical malfunction. AB - The role of intraluminal device pressure in the development of mechanical failures in 3-piece inflatable penile prostheses was investigated in a 2-part study. An in vitro study was performed in which mean intraluminal device pressures were recorded in Mentor IPP, Mentor Alpha-1, AMS 700 CX and AMS Ultrex devices. At maximum inflated volumes mean intraluminal device pressures exceeded 600 and 1,000 mm. Hg at rest, and 850 and 1,300 mm. Hg following external loading in Mentor and AMS products, respectively. The AMS 700 CX was associated with the highest mean intraluminal device pressure, with values approaching 1,400 mm. Hg following external loading. A 4-year clinical review at our institution was performed comparing the mechanical malfunctions in 51, 3-piece implants with connectors between the pump and cylinders (AMS 700 CX) to the mechanical malfunctions of 50, 3-piece implants with pre-connected tubing between the pump and cylinders (Mentor Alpha-1). Fluid leaks were identified in 16% of the former and 4% of the latter devices. It is proposed that the high intraluminal device pressures are an important factor in the pathophysiology of the mechanical malfunctions in 3-piece inflatable penile prostheses and that connectors are considered components at risk for pressure-associated fluid leaks. The ideal penile prosthesis should maintain the superior rigidity and cosmetic features of the 3-piece inflatable penile prosthesis. However, the device should be pre connected and designed to mimic more closely the anatomy of the human penis to achieve functional erections at lower intraluminal device pressure values. PMID- 8426406 TI - Efficiency and side effects of intracavernous injections of moxisylyte in impotent patients: a dose-finding study versus placebo. AB - We assessed the efficiency and tolerance of the alpha-blocking agent moxisylyte in 2 double-blind studies versus placebo performed in 12 neurogenic patients with spinal cord lesions and in 61 patients presenting with either psychogenic impotence (30) or erectile dysfunction that was predominantly neither psychogenic, hormonal nor neurogenic (31). In each etiological group patients were randomized (according to latin square method) to receive 3 single doses (10, 20 and 30 mg.) of moxisylyte and a placebo. The erectile response was determined 5, 10, 15, 20 and 30 minutes after each injection. Whatever etiology of impotence and dosage tested, the erectile response induced by moxisylyte was significantly higher than the placebo-induced response. No difference occurred among the 3 doses. In 93% of the patients moxisylyte induced an erectile response, including tumescence in 6, partial rigidity in 16 and complete rigidity in 46. Thus, in 62 of 73 patients (85%) the drug allowed initiation of erection adequate for intercourse. Placebo induced such erection in only 25% of the cases and in 55% there was no response. Tolerance was good and no priapism occurred. Only 4 patients (5%) reported mild pain during injection but erections were never painful, 1 presented with moderate and transient hypotension at the 20 mg. dose and a painless prolonged erection was observed in 1 case after the lowest dose. Drugs such as moxisylyte should be given before less well tolerated drugs. PMID- 8426407 TI - Penile venous ligation in 18 patients with 1 to 3 years of followup. AB - We report the results of penile venous ligation in 18 consecutive patients with erectile insufficiency due to corporeal venous occlusive dysfunction. Patient age ranged from 25 to 65 years (mean 47). Duration of erectile dysfunction ranged from 9 to 468 months (mean 95) and followup ranged from 12 to 37 months (mean 24). Of 18 patients 11 (61%) have sufficient persistent improvement in erections to permit unaided coitus. Of 7 failures 6 had temporary improvement in erections after the procedure: in 5 the improvement lasted 6 months or less, while only 1 had lasting improvement (24 months) before relapse. Reports of results of penile venous ligation should not include patients who have been followed for less than 12 months. Longer followup is needed before results of penile venous ligation beyond 2 years are known. PMID- 8426408 TI - Prostate size and configuration in adults with bladder exstrophy. AB - A total of 13 men born with classical bladder exstrophy underwent magnetic resonance imaging examination of the pelvis to evaluate the size and configuration of the prostate and pelvic organs. Mean patient age was 25.2 years (range 19 to 38). Of the patients 4 are voiding per urethram and 9 have undergone urinary division. Measurements included prostate volume, weight and maximum axial cross sectional area; pubic diastasis, and seminal vesicle size. The mean prostatic cross sectional area was 10.1 +/- 3.4 cm.2. The mean estimated prostatic volume and weight were 20.7 +/- 8.2 cc and 21.7 +/- 8.6 gm., respectively. The mean seminal vesicle length and width were 2.1 +/- 0.99 and 1.1 +/- 0.38 cm., respectively. The volume, weight and maximum cross sectional area of the prostate appear normal compared to published norms. In none of the patients did the prostate extend circumferentially around the urethra and the urethra was anterior to the prostate in all patients. Also, the puborectalis muscle group was widely separated and only provided lateral support of the prostate in patients who were continent and who had undergone prior posterior iliac osteotomy. Thus, the attainment of continence in this complex group of patients is multifactorial and prostate growth as evaluated by magnetic resonance imaging may not influence continence in these patients. PMID- 8426409 TI - Evaluation of current practices in routine preoperative crossmatching for transurethral resection of the prostate. AB - Increasing demand for blood during elective surgery has led to attempts to rationalize routine preoperative crossmatching to those operations when there is a reasonable (greater than 30%) chance of it being required. Results of a questionnaire returned by 86% of the urology units in Great Britain show that 41% continue to crossmatch 2 units of blood before transurethral prostatectomy. A review of 301 transurethral prostatectomies performed with preoperative grouping and saving of blood showed that only 11% of the patients required transfusion. The need for transfusion was significantly associated with presentation in chronic or acute on chronic retention (p < 0.01), and the operative factors of weight of resection (p < 0.0001) and degree of hemostasis (p < 0.001). Therefore, a policy of routine preoperative blood grouping and saving is safe, and could be widely adopted, leading to financial saving and a decrease in wastage of blood that becomes too old to use. PMID- 8426410 TI - An adequate sampling of the prostate to identify prostatic involvement by urothelial carcinoma in bladder cancer patients. AB - The distribution of any involved prostatic urethra, ducts and acini by urothelial carcinoma was studied to determine an adequate sampling method for detecting prostatic involvement using the maps of 38 cystoprostatectomy specimens. A total of 31 patients had prostatic duct and acini involvement, while 7 had prostatic urethral involvement alone. However, the distribution of the involved prostatic urethra, ducts and acini varied. In 29 of the 31 patients (93.5%) with prostatic duct and acini involvement, urethral carcinoma in situ and/or superficial gland involvement (an involvement of the afferent ducts within a few millimeters of the urethral mucosa) at the 5 and/or 7 o'clock position of the verumontanum portion was identified. In 7 patients with prostatic urethral involvement alone 2 had carcinoma foci at the 5 and/or 7 o'clock position of the verumontanum portion. Furthermore, the frequency of deeper gland involvement (an involvement of true prostatic acini except for superficial glands) was higher in patients with superficial gland involvement at the 5 and/or 7 o'clock position of the verumontanum portion (57.7%) than in patients without such involvement (20.0%). Therefore, this study emphasizes that a transurethral resection biopsy containing prostatic tissue at the 5 and/or 7 o'clock position of the verumontanum portion substantially improves the detection of prostatic duct and acini involvement in bladder cancer patients. Moreover, if the prostatic superficial glands are involved at the 5 and/or 7 o'clock position of the verumontanum portion, the potential involvement of the deeper glands should also be suspected. PMID- 8426411 TI - Complications of laparoscopic pelvic lymph node dissection. AB - Intraoperative and postoperative complications were assessed in the first 372 patients undergoing laparoscopic pelvic lymph node dissection at 8 medical centers. In 16 patients laparoscopic node dissection could not be completed due to patient body habitus or technical difficulties. Of these aborted procedures 14 occurred during the initial 8 dissections at each institution. A total of 55 complications (15%) occurred: 14 were noted in the intraoperative and 41 in the postoperative period. Of these patients 13 required open surgical intervention for the treatment of a complication. Complications included vascular injury (11 patients), viscus injury (8), genitourinary problems (10), functional/mechanical bowel obstruction (7), lower extremity deep venous thrombosis (5), infection/wound problem (5), lymphedema (5), anesthetic complications (2) and obturator nerve palsy (2). Based on our experience, there is a significant learning curve associated with performing laparoscopic pelvic node dissection. However, with experience and adherence to laparoscopic surgical principles, the risk of complications may be minimized. PMID- 8426412 TI - Deferred treatment of low grade stage T3 prostate cancer without distant metastases. AB - Fifty patients with extracapsular prostate cancer without known distant metastases were included in a protocol of surveillance followed by deferred treatment of progression. The median observed followup time was 78 months. The risk of developing distant metastases, if not dying before, was 24% and 37% at 5 and 9 years, respectively. The risk of dying of prostate cancer, if not dying of other diseases before, was 12% and 30% at 5 and 9 years, respectively. Thus, the cancer specific survival was 88% at 5 years and 70% at 9 years. PMID- 8426413 TI - Spinal cord compression secondary to prostate carcinoma: treatment and prognosis. AB - Of 35 patients with prostate carcinoma and suspected spinal cord compression 26 (74%) had myelograms and/or magnetic resonance imaging studies demonstrating epidural spinal cord compression. In 5 of 26 patients (19%) spinal cord compression was the first indication of prostate cancer. All patients were initially treated with radiation, steroids and androgen deprivation therapy. Three patients underwent laminectomy. Of 12 patients (100%) ambulatory at presentation 12 remained ambulatory. Of 12 patients (83%) who were paraparetic at presentation 10 were ambulatory after treatment. However, 2 of these patients subsequently had recurrent compression and became paraplegic. Overall, 7 of 26 patients (27%) had recurrent compression. Of 5 patients who either presented with paraplegia or in whom paraplegia developed secondary to recurrent spinal cord compression 4 remained paraplegic despite treatment. The average survival of these 5 patients after treatment was 3.9 months versus 18 months for the group as a whole. In ambulatory or paraparetic patients radiation, androgen deprivation therapy and steroids are effective palliative therapy. However, patients who present with paraplegia or in whom paraplegia developed secondary to recurrent compression are often not palliated by this combination therapy. Prophylactic radiation of vertebral metastases discovered concurrently with compressive metastases may be valuable in preventing paraplegia. PMID- 8426414 TI - Replacement of nephrostomy tube using ureteroscopes. AB - Replacement of a percutaneous nephrostomy tube can be a difficult task. The often time-consuming procedure involves replacement of a guide wire blindly or under fluoroscopic control, or by using ultrasound after decompression of hydronephrosis. A rigid 8.5F ureteroscope was used successfully to replace nephrostomy tubes in 3 patients with obstructive uropathy secondary to prostate carcinoma and in 1 with an ileoureteral anastomotic leak following radical cystectomy. PMID- 8426415 TI - Topographic-anatomical basis of sacral neurostimulation: neuroanatomical variations. AB - In 4 male and 5 female cadavers we undertook a detailed anatomical dissection of the caudal segment of the spinal cord with special emphasis on the conus medullaris, length of the spinal segment and pattern of nerve bridging between adjacent sacral roots. We found an unexpectedly high incidence of intradural bridging--some of simple formation and others more complex. These interconnections were more frequent between dorsal than ventral roots but never joined dorsal to ventral roots or vice versa. Ventral S5 was missing in some cases and we found no coccygeal roots. The dural sac was seen to extend to the S1 to S2 intervertebral disk or the lower margin of S2, permitting a limited sacral laminectomy to expose the intradural sacral roots in their entirety. Towards the end of the dural sac the ventral and dorsal roots were joined by an extremely thin epineurium, and each root was composed of several lightly fused rootlets that could be separated under magnification but that combined to constitute the root nerve bundle. Our study suggests that the sacral spinal cord is neither symmetrical nor metameric. It exhibits more numerous interconnections and variations than the rest of the spinal roots, which is consistent with the greater incidence of congenital malformations affecting this segment and the varying responses found with neurostimulation. PMID- 8426416 TI - Bilateral testicular adrenal rests in a patient with 11-hydroxylase deficient congenital adrenal hyperplasia. AB - A 13-year-old boy was referred for investigation of hypertension, precocious puberty and giant testicles. Hormonal studies established the diagnosis of congenital adrenal hyperplasia due to 11-hydroxylase deficiency. Testicular biopsy revealed that the scrotal masses were adrenal rest tumors entirely composed of adrenocortical tissue. Primary testicular tissue was absent. The size of the tumors regressed following replacement steroid therapy together with restoration of normal blood pressure. This observation of a testicular adrenal rest tumor in a patient with 11-hydroxylase deficient congenital adrenal hyperplasia is unique. PMID- 8426417 TI - Testis sparing surgery for pediatric epidermoid cysts of the testis. AB - Epidermoid cyst of the testis is a rare, benign intratesticular tumor in children. Four patients 4 to 15 years old with epidermoid cysts were managed by tumor excision with salvage of the testis. Preoperative ultrasound, tumor marker status and intraoperative findings were suggestive of a benign neoplasm. Under these circumstances epidermoid cysts may be safely treated by excision in prepubertal patients. PMID- 8426418 TI - Infantile fibromatosis of the external genitalia: diagnosis and management strategy. AB - Infantile fibromatosis is a process that is difficult to distinguish from fibrosarcoma. It is found in neonates and infants but only rarely has it been reported to involve the external genitalia or scrotum. A 10-month-old boy is described with histologically proved infantile fibromatosis of the scrotum. Characteristics distinguishing infantile fibromatosis from fibrosarcoma and the appropriate management of this disease are presented. PMID- 8426419 TI - Use of extracorporeal shock wave lithotripsy in a solitary kidney with renal artery aneurysm. AB - Arterial aneurysms have been among the relative contraindications cited for extracorporeal shock wave lithotripsy. We report the successful use of extracorporeal shock wave lithotripsy for treatment of renal calculi in a solitary kidney with renal artery aneurysms. Appropriate management considerations and perioperative precautions are discussed. PMID- 8426420 TI - Delayed allograft autotransplantation after excision of a large symptomatic renal artery pseudoaneurysm. AB - Pseudoaneurysm formation, which occurs infrequently in the renal transplant patient, can have a catastrophic outcome. We report a case of a large, symptomatic transplant renal artery pseudoaneurysm that was treated by excision with extracorporeal vascular repair and delayed allograft autotransplantation. We review the management of transplant renal artery pseudoaneurysms and the role of delayed autotransplantation in complex reconstructive renal surgery. PMID- 8426421 TI - Angiosarcoma arising from a defunctionalized arteriovenous fistula. AB - We report a case of angiosarcoma developing in a defunctionalized arteriovenous fistula that had been created for hemodialysis approximately 20 years previously. At presentation the patient had a 30-year history of chronic renal failure and had undergone renal transplantation 10 years previously. PMID- 8426422 TI - Pyeloureterostomy in the management of renal allograft ureteral complications: an alternative technique. AB - Pyeloureterostomy is the standard procedure for reconstructing renal allograft ureteral complications. Most reports describe an end-to-end technique with or without native nephrectomy. An alternative is an end-to-side anastomosis, leaving the native ureter in continuity. We report our experience with the latter method. Since July 1983, 437 renal transplantations have been performed at our institution. End-to-side pyeloureterostomy has been used in 5 cases for urological reconstruction after renal transplantation following ureteral ischemic necrosis or stenosis. In 1 patient the native kidneys had been removed several years previously but in the remaining 4 the native kidneys were left in situ. There have been no significant complications following this procedure. We believe that by not significantly mobilizing, ligating or dividing the native ureter the chance of anastomotic breakdown due to ischemia may be decreased. PMID- 8426423 TI - Intermittent self-catheterization of the renal pelvis: report of 2 cases. AB - We review the long-term outcome of 2 patients in whom cutaneous ureterostomies were performed. Complications included necrosis and distal ureteral stenosis, peristaltic dysfunction, urosepsis, calculus formation and renal impairment. Fluoroscopic ureterometry confirmed high pressure collecting systems in both patients 14 to 17 years postoperatively. Subsequent management during the last 4 to 6 years with clean intermittent self-catheterization has resulted in a significant improvement in the urological status. PMID- 8426424 TI - Urethrocavernous fistula following sexual intercourse. AB - An unusual case of urethrocavernous fistula following blunt penile trauma sustained during sexual intercourse is described. The fistula was well outlined by cavernosography and responded successfully to conservative treatment. PMID- 8426425 TI - Treatment of penile incarceration by the string method: 2 case reports. AB - We report 2 cases of penile incarceration in a 10-year-old boy and a 40-year-old man. In both cases the penis was encircled by a metallic object. The string method described was successful, rapid, atraumatic and simple to use. PMID- 8426426 TI - Re: Iohexol clearance for the determination of glomerular filtration rate in clinical practice: evidence for a new gold standard. PMID- 8426427 TI - Re: Bladder outlet obstruction versus impaired detrusor contractility: the role of uroflow. PMID- 8426428 TI - Re: Venous leaks: anatomical and physiological observations. PMID- 8426429 TI - Re: Functional characteristics of sperm obtained by electroejaculation. PMID- 8426430 TI - The effects of a soft tissue mimicking medium and increased power settings on the location and magnitude of lithotripter peak positive pressure. AB - In vitro experiments showed that a tissue mimicking medium alters the peak positive pressure (p+), focal zone properties and frequency content of shockwaves compared with their behavior in water. The reduction in (p+) ranged from 5% at 10 kV. to 19% at 18.1 kV., when measured at the geometric focus with the tissue mimicking medium present. As power settings were increased, the relative gain in pressure was damped by attenuation. A 2 mm. shift in the acoustic focus was seen both axially and laterally with the tissue mimicking medium. While the former is probably not significant, the latter may be clinically significant given the narrow lateral beam width at the acoustic focus. These attenuation experiments suggest that clinical targeting through tissue may not be as precise or result in as high peak pressures as the clinician expects. Especially at high power settings, the depth of tissue traversed should be minimized to limit attenuation effects. PMID- 8426431 TI - Response of the human urinary bladder to angiotensins: a comparison between neurogenic and control bladders. AB - The response of the human detrusor muscle to angiotensins was investigated and compared between neurogenic and control bladders. Both angiotensin I and II induced potent contraction of the human detrusor muscle. Saralasin completely inhibited the response to both angiotensins, while verapamil and indomethacin barely suppressed the contractility provoked by angiotensin II. Captopril completely blocked the response to angiotensin I. The contractile response of angiotensin II was abolished in Ca(++)-free Krebs' solution. The contractile strength of the neurogenic bladders induced by both angiotensin I and II was significantly weaker than that of the controls. However, there was no difference in ED50 value between the 2 groups. These results support the hypothesis that angiotensin I is converted to angiotensin II by angiotensin converting enzyme in the detrusor, and that angiotensin II subsequently contracts the detrusor muscle through angiotensin II receptors. The bladder contractility induced by the angiotensins was significantly less potent in the neurogenic bladders than in the control. PMID- 8426433 TI - First gene therapy for inherited hypercholesterolemia a partial success. PMID- 8426434 TI - Researchers try new definitions, new therapies in effort to solve growing problem of sepsis. PMID- 8426432 TI - Magnesium oxide administration and prevention of calcium oxalate nephrolithiasis. AB - We studied the effect of oral administration of magnesium oxide (MgO) on calcium oxalate (CaOx) nephrolithiasis in rats. Nephrolithiasis was induced by administration of 1.0% ethylene glycol (EG) in drinking water. Magnesium oxide was given mixed with food at 500 mg./100 g. rat chow. Dispensation of MgO resulted in a significant increase of urinary pH and a modest increase in urinary excretion of citrate. Urinary excretion of oxalate started to decline by day 14 and was significantly reduced on days 21 and 28. All rats receiving EG displayed crystalluria. From the group receiving EG only, 3 of 4 rats sacrificed on day 15 and 2 of 4 rats sacrificed on day 29 had CaOx crystal deposits in their kidneys. None of the 8 rats who received both EG and MgO had CaOx nephrolithiasis. Thus our findings indicate that dispensation of magnesium as MgO can be beneficial against calcium oxalate nephrolithiasis. PMID- 8426435 TI - 'Desperate use' gene therapy guidelines ready. PMID- 8426436 TI - From the Congressional Office of Technology Assessment. PMID- 8426437 TI - From the Centers for Disease Control and Prevention. Hepatitis E among US travelers, 1989-1992. PMID- 8426438 TI - From the Centers for Disease Control and Prevention. International notes: surveillance of health status of Bhutanese refugees--Nepal, 1992. PMID- 8426439 TI - From the Centers for Disease Control and Prevention. Deaths attributed to nor'easter--December 1992. PMID- 8426440 TI - From the Centers for Disease Control and Prevention. Preliminary data: exposure of persons aged > or = 4 years to tobacco smoke--United States. PMID- 8426442 TI - Quality of care in rural hospitals. PMID- 8426441 TI - Quality of care in rural hospitals. PMID- 8426443 TI - DRG miscoding: error or intent? PMID- 8426444 TI - Electromagnetic fields and circadian rhythms. PMID- 8426445 TI - Ecstasy, the serotonin syndrome, and neuroleptic malignant syndrome--a possible link? PMID- 8426446 TI - A prospective cohort study of vasectomy and prostate cancer in US men. AB - OBJECTIVE: To examine prospectively the relationship between vasectomy and prostate cancer. DESIGN: Cohort study. SETTING: Health professionals (dentists, veterinarians, osteopaths, optometrists, pharmacists, and podiastrists) in the United States. PARTICIPANTS: There were 10,055 male members of the Health Professionals Follow-up Study, aged 40 to 75 years, who had had a vasectomy, and 37,800 members who had not had a vasectomy at the time of study entry in 1986. These participants had provided detailed information on various life-style variables including diet. MAIN OUTCOME MEASURE: Diagnosis of prostate cancer. RESULTS: Between 1986 and 1990, 300 new cases of prostate cancer were diagnosed in participants who were initially free of diagnosed cancer. Vasectomy was associated with an elevated risk of prostate cancer (age-adjusted relative risk, 1.66; 95% confidence interval, 1.25 to 2.21; P = .0004). This elevated risk persisted after excluding 21 stage A1 cases (age-adjusted relative risk, 1.56; 95% confidence interval, 1.15 to 2.11; P = .004). Among men who had their vasectomy at least 22 years in the past (before 1965), the risk of prostate cancer was even higher (relative risk, 1.85; 95% confidence interval, 1.26 to 2.72; P = .002). This elevated risk among men with vasectomy did not appear to be caused by detection bias and persisted when we controlled for diet, level of physical activity, smoking, alcohol consumption, educational level, body mass index, and geographical area of residence. CONCLUSIONS: These results support evidence from other epidemiologic studies that vasectomy increases risk of prostate cancer. The consistency of results among various epidemiologic studies, the increase of risk over time following vasectomy, the apparent lack of confounding or bias, and the existence of physiological changes in the prostate following vasectomy suggest that the association may be causal. PMID- 8426447 TI - Transmission of Escherichia coli O157:H7 infection in Minnesota child day-care facilities. AB - OBJECTIVE: Escherichia coli O157:H7 infection can cause hemorrhagic colitis and hemolytic uremic syndrome. Since 1988 the Minnesota Department of Health has carried out surveillance for this infection. To assess the occurrence of person to-person transmission within day-care facilities, we investigated facilities where an infected child attended after onset of symptoms. DESIGN: Parents of children less than 5 years old with E coli O157:H7 infection were interviewed from July 1988 through December 1989. If the child attended day care after onset, stool cultures were obtained from other children in attendance and their parents were interviewed. If there was presumptive evidence of ongoing E coli O157:H7 transmission in a facility, all preschool children were excluded from attending day-care facilities until two consecutive stool cultures were negative. RESULTS: Sixty-eight cases of E coli O157:H7 infection were identified in Minnesota during the 18-month period, including 29 cases identified through investigations at nine day-care facilities. There was evidence of person-to-person transmission in all nine facilities. The median number of infected children per facility was two (range, two to 18), and the median attack rate was 22% (range, 3% to 38%). The median age of the primary case at each facility was 26 months (range, 12 to 59 months). There was no evidence of further transmission at facilities where children were temporarily excluded until two consecutive stool cultures were negative. CONCLUSION: Person-to-person transmission of E coli O157:H7 is common when infected preschool children attend day care while symptomatic. The number of unrecognized day-care outbreaks in the United States may be substantial due to the lack of routine testing for this pathogen in stool cultures, the absence of public health surveillance in many regions, and incomplete follow-up of infected children. Temporary exclusion of all children was an effective control strategy in this population, but additional investigations are needed to determine the optimal intervention. PMID- 8426448 TI - Insuring Latinos against the costs of illness. AB - OBJECTIVE: To examine the determinants of health insurance coverage for Latinos in the United States and how different targeted strategies for health care reform differentially affect the country's major ethnic groups, focusing on the implications for the Latino population. DESIGN: Data from the 1980 and 1990 Current Population Surveys were used to compare the insurance status of nonelderly (< 65 years) Latinos with the Anglo (non-Hispanic white), black, and Asian and other populations by estimating the attributable fraction for selected covariates. The effects of health care reform strategies on the coverage of the major ethnic groups were simulated from these data. MAIN OUTCOME MEASURES: Percentage uninsured, percentage insured by Medicaid, and attributable fraction for covariates. RESULTS: Latinos have the worst health insurance coverage of any ethnic group in the country. Approximately 39% of Latinos are uninsured compared with 13.8% for the Anglo and 24% for the black population. Providing coverage to all the poor could reduce the uninsured rate for Anglos by about 23%, whereas the reduction among Latinos could be about 37% and among blacks about 42%. Similar reductions could be achieved by covering all workers and their minor dependents. Regardless of the approach to reform, however, Latinos would remain with high absolute rates of uninsured. CONCLUSIONS: Differences in Medicaid eligibility, labor force characteristics, and family composition between Latinos and other ethnic groups suggest that policy initiatives may affect Latinos differently. Targeted strategies, such as employer mandates, "pay-or-play" programs, or Medicaid expansions, can improve coverage, but many Latinos could still remain uninsured. PMID- 8426449 TI - Vitamin A supplementation and child mortality. A meta-analysis. AB - OBJECTIVE: A two-part meta-analysis of studies examining the relationship of vitamin A supplementation and child mortality. DATA SOURCES: We identified studies by searching the MEDLARS database from 1966 through 1992 and by scanning Current Contents and bibliographies of pertinent articles. STUDY SELECTION: All 12 vitamin A controlled trials with data on mortality identified in the search were used in the analysis. DATA EXTRACTION: Data were independently extracted by two investigators who also assessed the quality of each study using a previously described method. DATA SYNTHESIS: We formally tested for heterogeneity across studies. We pooled studies using the Mantel-Haenszel and the DerSimonian and Laird methods and adjusted for the effect of cluster assignment of treatment groups in community-based studies. Vitamin A supplementation to hospitalized measles patients was highly protective against mortality (DerSimonian and Laird odds ratio, 0.39; 95% confidence interval, 0.22 to 0.66; P = .0004) (part 1 of the meta-analysis). Supplementation was also protective against overall mortality in community-based studies (DerSimonian and Laird odds ratio, 0.70; clustering adjusted 95% confidence interval, 0.56 to 0.87; P = .001) (part 2 of the meta analysis). CONCLUSIONS: Vitamin A supplements are associated with a significant reduction in mortality when given periodically to children at the community level. Factors that affect the bioavailability of large doses of Vitamin A need to be studied further. Vitamin A supplements should be given to all measles patients in developing countries whether or not they have symptoms of vitamin A deficiency. PMID- 8426450 TI - Criteria for evaluating a ban on the advertisement of cigarettes. Balancing public health benefits with constitutional burdens. PMID- 8426451 TI - Foods of the future. The new biotechnology and FDA regulation. PMID- 8426452 TI - Scientific misconduct. New definition, procedures, and office--perhaps a new leaf. PMID- 8426453 TI - A piece of my mind. The ear. PMID- 8426454 TI - Diagnostic and therapeutic technology assessment. Lung transplantation. PMID- 8426455 TI - Prediction of elder caregiver burden. PMID- 8426456 TI - Antibiotic-associated pseudomembranous colitis with toxic megacolon after cesarean section. PMID- 8426457 TI - Base deficit as a guide to injury severity and volume resuscitation. AB - Base deficit is an expeditious and sensitive measure of both the degree and duration of inadequate perfusion, and has utility in the clinical evaluation and management of victims of trauma. It is rapid, accurate, and widely available, and as such may be very useful in the initial evaluation of these patients; it may also be used to help guide and assess the adequacy of volume resuscitation. It has in addition both clinical value and research potential in predicting outcome in these patients, and may be a useful tool to both the clinician and the researcher. PMID- 8426458 TI - Fever and rash in a young man [corrected]. PMID- 8426459 TI - An unusual pneumonia. PMID- 8426461 TI - Loss prevention case of the month. Who's my doctor? PMID- 8426460 TI - HIV reporting in Tennessee: impact and trends. PMID- 8426462 TI - Tennessee physicians join national effort to end domestic violence. PMID- 8426463 TI - In a heartbeat. Interview by David E. Reiser. PMID- 8426465 TI - Comparison between lung parenchyma and bronchoalveolar lavage collagenolytic activity. AB - We have evaluated, in an experimental model of silicosis in guinea pigs, if the presence of collagenolytic activity in bronchoalveolar lavage (BAL) fluid reflects the collagen catabolism in lung parenchyma. We measured simultaneously BAL collagenase activity, using as substrate [3H]type I collagen, and lung collagenolytic activity by the tissue pellet assay. Animals (n = 30) were instilled intratracheally with 50 mg of quartz DQ-12 and sacrificed 15, 30, and 60 days after silica administration. Guinea pigs instilled with saline solution were used as controls. Our results showed that lung parenchymal collagenolytic activity was present in all experimental and normal guinea pigs. There were no statistical differences between silicotic and normal animals at 15 and 30 days. At 60 days, however, a significant decrease in tissue collagenolytic activity was observed in silicotic animals (161 +/- 100 vs. 400 +/- 152 units of collagenase activity; p < 0.001). In contrast, BAL collagenolytic activity was revealed only in 7 of 10 silicotic animals at 15 days and 30 days, and in 4 of 10 at 60 days. Normal guinea pigs did not exhibit BAL collagenase activity. BAL and tissue collagenase activity from each experimental animal were analyzed by straight line regression and no significant relationship was observed (r = 0.082; p = 0.87). This suggests that BAL collagenolytic activity does not reflect lung tissue collagen turnover. PMID- 8426464 TI - Surfactant-associated proteins (SP-A, SP-B) are increased proportionally to alveolar phospholipids in sheep silicosis. AB - In silicosis, a profile of surfactant overproduction associated with type II epithelial cell hypertrophy and hyperplasia has been documented. In addition, enhanced accumulation of surfactant-associated protein A (SP-A) has been seen in the rat model of acute silicosis by 2 independent groups, but the exact role of these surfactant changes in silicosis are incompletely understood. In this study we measured in lung lavage surfactant total phospholipids and surfactant associated proteins A and B (SP-A, SP-B). In addition, the surface tension reducing activities of lipid extracts of bronchoalveolar lavage fluids (BALFs) in the sheep silicosis model were examined using a pulsating bubble surfactometer. Two groups of animals (n = 18) were investigated: a saline-exposed (PBS-PBS) group and a silica-exposed (Si-PBS) group. Total surfactant phospholipids in the silicotic sheep increased 2-fold over the control sheep values (p < 0.05), as previously reported. In addition, we found a substantial rise in total surfactant associated proteins, with significant increase in SP-A (1.16 +/- 0.22 micrograms/ml Si-PBS group vs. 0.70 +/- 0.07 micrograms/ml PBS-PBS group, p < 0.05) and a parallel but not significant increase in SP-B (2.68 +/- 0.90 micrograms/ml Si-PBS group vs. 1.10 +/- 0.30 micrograms/ml PBS-PBS group). The surface-tension-reducing activities of alveolar fluid lipid extracts did not differ significantly between the groups (Si-PBS group at maximal bubble radius [Rmax]: 27.0 +/- 1.6 and at minimal bubble radius [Rmin]: 1.0 +/- 0.7 milli Newton/m, vs. PBS-PBS group Rmax: 27.0 +/- 2.2 and Rmin: 0.7 +/- 0.3 milli Newton/m at 4 min pulsation). The ratios of SP-A and SP-B to lipid phosphorus levels document a proportional enhancement of surfactant-associated proteins and phospholipids, thus suggesting a co-ordinated upregulation of both surfactant associated proteins and phospholipids in this model of silicosis. However, on an individual basis, these changes were not related, suggesting a more complex model of regulation. This study documents significant increases of the surfactant apoproteins proportional to changes in phospholipids in the lung of silicotic sheep. In spite of these alterations of surfactant components, organic solvent lipid extracts of BAPLFs surfactant remained equally effective in reducing surface tension. PMID- 8426466 TI - Airway hyperresponsiveness in cigarette smoke-exposed rats. AB - To investigate the possibility that altered airway-parenchymal interaction may account for bronchial hyperresponsiveness induced by cigarette smoke exposure, we tested the effect of administration of cigarette smoke (SM), elastase (EL), and both SM and EL on airway responsiveness in 41 Long-Evans male rats. Twelve were exposed to 30 puffs of SM for 15 weeks; 8 received a single intratracheal injection of EL (250 IU/kg); 9 received both EL and SM exposure (SE); 12 control rats were exposed to room air (CO). After 15 weeks, animals were anesthetized and mechanically ventilated (Vt = 2.5 ml, f = 80/min). Methacholine (MCh) dose response curves (DRCs) were constructed by calculating pulmonary resistance (RL) after ultrasonic nebulization of saline followed by doubling concentrations of MCh (0.0625-256 mg/ml). Exposure to cigarette smoking, with or without elastase, led to a significant reduction in body weight and increased total lung capacity (TLC) compared to exposure to CO. However, there was no significant change in static compliance in the experimental groups, despite increased lung volume. The concentration resulting in a doubling of RL (EC200RL) was significantly lower in rats treated with SM (n = 7) than CO (n = 8) (3.3 vs. 56.1 mg/ml, geometric mean, p < 0.01). The concentration at which a maximal RL was achieved was lower in SM than CO, EL, and SE (p < 0.05). To assess the possible influence of airway parenchymal interaction on responsiveness, we measured RL both at functional residual capacity (FRC) and at a volume above FRC equivalent to 1 tidal volume. RL changed similarly in all groups. Despite similar effects on mechanics of both cigarette smoke exposure and elastase administration, only cigarette smoke exposed animals exhibited evidence of hyperresponsiveness. In this model cigarette smoke-induced hyperresponsiveness is unrelated to changes in either lung elasticity or airway-parenchymal interaction. PMID- 8426468 TI - Cytogenetic heterogeneity in t(11;19) acute leukemia: clinical, hematological and cytogenetic analyses of 48 patients--updated published cases and 16 new observations. AB - We report on 16 cases of t(11;19) acute leukemia and review data of published observations: altogether updated data of 48 patients are analyzed. Four hematological groups could be distinguished: (i) 13 cases of acute lymphoblastic leukemia (ALL) of B lineage, mostly CD19+; (ii) eight cases of biphenotypic leukemia: CD19+ (most often) ALL but with simultaneous or inducible expression of differentiation marker of monocytic lineage. The B lineage and biphenotypic leukemias were predominantly found in female infants; (iii) four cases of T-ALL in children; and (iv) 23 acute non-lymphocytic leukemia (ANLL) cases generally of M4 or M5 subtype, predominantly in males. Cytogenetically, at least two subtypes were observed with possibly an identical breakpoint on 11q23 but discrete breakpoints on 19p: lymphoid, biphenotypic, and most congenital myeloid cases showed a distal breakpoint on 19p13 producing 11q- and 19p+ derivatives, while most older myeloid cases showed 11q+ and 19p- derivatives as a result of a more proximal breakpoint on 19p12 or p13.1. The latter type was clearly detected using R bands but barely visible using Q or G bands while the other translocation was easy to detect with G bands but could be missed with R bands. The white blood cell count is usually high in these t(11;19) acute leukemias and prognosis is poor, except for T-ALL cases. PMID- 8426467 TI - Prognostic implications of breakpoint and lineage heterogeneity in Philadelphia positive acute lymphoblastic leukemia: a review. AB - Philadelphia-positive (Ph) acute lymphoblastic leukemia (ALL) is heterogeneous both in terms of the BCR gene breakpoints (M-bcr and m-bcr) and in the number of cell lineages carrying the Ph chromosome. The impact of these observations on the controversy surrounding Ph ALL and Ph+ chronic granulocytic leukemia (CGL) is unclear. Twenty cases of Ph ALL (four newly investigated and 16 previously published) were classified into nine stem-cell (the Ph in myeloid and lymphoid cells) and 11 lymphoid-restricted cases. Lymphoid cases had a lower leucocyte count (56 x 10(9)/l) than stem-cell cases (151 x 10(9)/l) (p < 0.01). The M-bcr and m-bcr breakpoints (in 14 cases) were found in lymphoid (four cases of each) and stem-cell (five and one cases respectively) cases. Lymphoid cases had significantly shorter event-free survival (median 4 months) than stem-cell cases (median 35 months) (p < 0.01). M-bcr cases were older (mean 41 years) than m-bcr cases (mean 36 years)(p = NS); m-bcr cases had higher leukocyte counts (mean 85 x 10(9)/l) than M-bcr cases (36 x 10(9)/l)(p < 0.01) but breakpoint had no impact on prognosis. Lineage involvement, but not breakpoint, appears to distinguish prognostically important sub-groups of Ph ALL. Paradoxically, lymphoid-restricted cases have a worse prognosis than cases arising in a pluripotent stem-cell. PMID- 8426469 TI - Characteristics of pro-T ALL subgroups: comparison with late T-ALL. The Groupe d'Etude Immunologique des Leucemies. AB - A group of 30 acute lymphoblastic leukemias (ALL) with the early pro-T phenotype CD7+/cCD3+/CD1-/CD3-/CD4-/CD8-were identified among 103 newly diagnosed ALL with T-lineage markers (T-ALL). Pro T-ALL was more often observed in adults, and showed a lower incidence of hyperleukocytosis than more mature T-ALL. Mediastinal masses and polar acid phosphatase positivity in blast cells were however observed with the same frequency in pro T-ALL and late T-ALL, and rearrangements of both T cell receptor (TCR) beta and gamma genes were observed in half the pro T-ALL cases tested. The expression of CD34, DR, and myeloid (My) markers was significantly more frequent in pro T-ALL than in late T-ALL, and these three features were strongly linked. TCR gene rearrangements were two to three times more frequent in CD34- and My-pro T-ALL. However, both CD34+ and My+ pro T-ALL showed an incidence of mediastinal masses and polar acid phosphatase positivity similar to this observed in CD34- and My- cases. This supports the assumption that both types of ALL indeed are engaged in the T-lineage, and confirms intracytoplasmic cCD3 as the earliest marker for this lineage. Moreover, CD34 appears to persist up to an early stage of T-cell maturation, where the cells retain myeloid potentiality. Loss of CD34 correlates with TCR-beta gene rearrangement and definitive commitment to the T lineage. Event-free survival analysis suggested a poorer outcome for pro T-ALL in adult patients. PMID- 8426470 TI - Detection of residual proliferating leukemic cells by fluorescence in situ hybridization in CML patients in complete remission after interferon treatment. AB - Interferon-alpha produces a complete hematologic and cytogenetic remission in approximately 20% of patients with chronic myelogenous leukemia (CML). In this study, we applied fluorescent in situ hybridization (FISH) methodology to examine the possibility that a low level of proliferating Philadelphia-chromosome positive (Ph+) cells may be present in interferon-treated CML patients who have achieved complete cytogenetic remission (as defined by the absence of Ph chromosome in 20-25 metaphases analyzed). Ten such patients in remission for 6-35 months were studied by this technique, in which a chromosome-22-specific DNA painting probe was used to detect leukemic cells with the characteristic 9;22 chromosomal translocation. In six of the 10 patients (60%), 3-9% Ph+ metaphases were detected. No Ph+ cells were observed in nine control individuals. Thus, this study demonstrates that FISH technology is more sensitive than conventional cytogenetic analysis for the detection of minimal residual disease in CML. PMID- 8426471 TI - Predominance of a distinct subtype of hairy cell leukemia in Japan. AB - Forty Japanese patients with hairy cell leukemia (HCL) were reviewed. Nine cases were diagnosed as typical HCL, and two cases had the features of HCL variant (prolymphocytic variant). The remaining 29 cases (72.5%) differed morphologically and hematologically from the other two groups in that they usually had a moderately high leukocyte count (average 27.9 x 10(3)/microliters), and abnormal cells showing a densely stained round nucleus and an inconspicuous nucleolus. Tartrate-resistant acid phosphatase reaction was weak, and their cells exhibited generally smooth or slightly irregular, cellular outlines in smears. The cells showed weak expression of surface immunoglobulin G (IgG) with kappa-chain predominance. CD25 antigen was not detected. Some of these findings resemble those of B-cell chronic lymphocytic leukemia, but the patients also had several important features of HCL. They had splenomegaly without significant lymphadenopathy. The abnormal cells were CD20+, CD11c+ and showed typical 'hairy morphology' under phase-contrast and scanning electron microscopy. Furthermore, spleen sections revealed diffuse infiltration by the abnormal cells in the red pulp. From these findings, we speculated that this group of patients constitute a distinct subtype of HCL which is commonly seen in Japan. We propose to term the disease as HCL Japanese variant. PMID- 8426472 TI - Mini-dose interferon alpha-2a in the treatment of myelodysplasia. AB - Interferon-alpha has been used as a differentiating agent in the treatment of patients with myelodysplastic syndrome with conflicting results and often significant toxicity. In order to maximize the differentiating effects of this agent and minimize the myelosuppressive effects, a prospective pilot study was initiated utilizing interferon alpha-2a (Roferon A, Roche Laboratories) in the treatment of complicated or poor prognosis myelodysplasia. The study regimen utilized 'mini-dose' interferon alpha-2a at 1 x 10(6) units subcutaneously three times per week for 16 weeks followed by an 8 week observation period. Nine patients were enrolled between May 1990 and June 1991, of which seven are evaluable. Forty-three percent (3/7) had a partial or clinical response as defined by normalization of one or more of the hemoglobin concentration, white blood cell count, or platelet count, or a decrease in transfusion requirement by > or = 50%. Only one patient was removed from study for interferon-associated toxicity. Mini-dose interferon alpha-2a appears to be an effective regimen for some patients with myelodysplasia which can be administered with minimal toxicity. Further investigation with interferon-alpha for the treatment of myelodysplastic syndrome, at the dosage utilized in this study, is warranted. PMID- 8426473 TI - Idarubicin in combination with intermediate-dose cytarabine and VP-16 in the treatment of refractory or rapidly relapsed patients with acute myeloid leukemia. The GIMEMA Cooperative Group. AB - Ninety-seven patients with refractory or relapsed acute myelogenous leukemia (AML), median age 37 years, received as salvage therapy a single course of idarubicin 6 mg/m2 as an intravenous (i.v.) bolus daily for 5 days, cytarabine (Ara-C) 600 mg/m2 i.v. for a period of 2 hours daily for 5 days and etoposide (VP 16) 150 mg/m2 for a period of 2 hours daily for 3 days (ICE protocol). Thirty-six patients were primarily resistant to standard inductive therapy with daunorubicin and Ara-C; 50 patients were in first relapse, three patients in second or third relapse, and eight patients in relapse after transplants. Forty-two (43%) out of 97 patients achieved complete remission, 11 patients died of infection or hemorrhage during induction, and 44 patients (45%) had resistant disease. Of the various variables examined, only disease status (i.e. refractory versus relapsed AML) was predictive for a significantly lower response rate. The median remission duration was 16 weeks; the overall median survival was 10 weeks. Nausea, vomiting, and oral mucositis were common but were rarely severe. No patient experienced treatment-related cardiac toxicity. In conclusion, the ICE protocol is a tolerable regimen providing effective antileukemic activity in patients with advanced AML. The evolution of this protocol in previously untreated patients with AML appears indicated. PMID- 8426474 TI - Splenectomy for patients with myelofibrosis with myeloid metaplasia: pretreatment variables and outcome prediction. AB - Since according to the early studies, the outcome after splenectomy in the individual patient with myelofibrosis with myeloid metaplasia (MMM) is unpredictable, we assessed retrospectively the pre-intervention characteristics that best predicted adverse events, hematological consequences, and survival in 71 splenectomized MMM patients. The findings indicate that the operative risk of splenectomy for both mortality (8.4%) and morbidity (39.3%) was unpredictable. New hemorrhagic or thrombotic complications occurred in 16.9% of surviving patients and were predicted by age < 50 years, a normal to high platelet count (> 200 x 10(9)/l) and huge splenomegaly (> 16 cm from the costal margin). Massive liver enlargement occurred in 24% of patients and has to be expected in patients splenectomized for transfusion-dependent anemia. Anemia improved substantially in 45% and 52% of patients at 3 months and at 1 year, respectively, and was predicted by severe anemia, low platelet count (< 100 x 10(9)/l) or normal to high white blood cell (WBC) count (> 4 x 10(9)/l). Survival from splenectomy was superior in patients < 45 years with WBC < 10 x 10(9)/l count. An unexpectedly high rate of blastic transformation was observed. It accounted for 42.8% of the deaths. The results suggest trials for prophylactic cytoreductive treatment in young patients and when platelet count is normal to increased. Further study is needed for elucidating the possible role played by splenectomy in inducing blastic transformation. PMID- 8426475 TI - Suppression of chronic myelogenous leukemia colony growth by interleukin-4. AB - Interleukin-4 (IL-4) is a cytokine with pleiotropic activities. In normal bone marrow cultures grown in the presence of either granulocyte-macrophage colony stimulating factor (GM-CSF) or interleukin-3 (IL-3), IL-4 suppresses granulocyte macrophage colony-forming unit (CFU-GM) proliferation but it enhances the colony stimulatory effect of granulocyte colony-stimulating factor (G-CSF). We studied the effect of IL-4 on chronic myelogenous leukemia (CML) bone marrow or peripheral blood cells from 30 patients using the CFU-granulocyte-erythrocyte monocyte-megakaryocyte colony culture assay. In several repetitive experiments, IL-4 inhibited CFU-GM colony replication by 24 to 65% in a dose-dependent fashion at concentrations ranging from 0.01 to 10 micrograms/ml when patients' cells were cultured in the presence of erythropoietin alone or with phytohemagglutinin conditioned medium, GM-CSF, or IL-3. The addition of 100 U/ml of IL-1 beta to the CML cultures partially reversed the inhibitory effect of IL-4. Incubation of CML low-density peripheral blood cells with IL-4 resulted in down-regulation of IL-1 beta and IL-6 production in three of four samples, suggesting that the suppressive effect of IL-4 is mediated by inhibition of IL-1 and by other mechanisms including inhibition of IL-6 production. In contrast to the stimulatory effect exerted by IL-4 on G-CSF-dependent CFU-GM progenitor proliferation in normal marrow, the addition of IL-4 to CML cultures grown in the presence of G-CSF resulted in a divergent effect: suppression of CML CFU-GM in two, stimulation in three, and no significant effect in two CML patients' samples. It is therefore possible that IL-4 may have an in vivo antiproliferative effect in a subpopulation of CML patients. PMID- 8426476 TI - Interleukin-6 receptor expression and saturation on the bone marrow cells of patients with multiple myeloma. AB - Expression of the interleukin-6 (IL-6) receptor on B cells and plasma cells in the bone marrow (n = 18) and peripheral blood (n = 32) of patients with multiple myeloma and the relative saturation of these receptors with endogenous IL-6 has been determined by dual-labelled flow cytometric analyses. B cells were identified using an anti-CD19 monoclonal antibody and plasma cells were identified by gating on cells with high fluorescent staining with anti-CD38. With the exception of one patient, very few bone marrow plasma cells expressed the IL 6 receptor (IL-6R) (mean = 2%). This was in contrast to cells from the U-266 plasma cell line, 90% of which had IL-6R. IL-6R expression was lower on bone marrow B cells (mean = 11%) than on peripheral blood B cells (mean = 69%). Studies using either monoclonal or polyclonal anti-human IL-6 to detect endogenous receptor-bound IL-6 found that the IL-6R on bone marrow B cells and plasma cells from patients with multiple myeloma were not saturated with endogenous IL-6 and the presence of receptor-bound IL-6 tended to be associated with stable disease. Thus dysregulated IL-6R expression was not evident on the B cells and plasma cells of patients with multiple myeloma and the increased IL-6R expression on the U-266 plasma cell line was not found on patients' cells. PMID- 8426477 TI - Chromosome 11 translocation breakpoints at the PRAD1/cyclin D1 gene locus in centrocytic lymphoma. AB - Centrocytic lymphoma is a CD5-positive B-cell neoplasm. Rearrangements at the chromosome 11q13 bcl-1 breakpoint loci are present in the majority of these lymphomas, as a result of reciprocal translocation with the 14q32 immunoglobulin heavy chain joining genes. Recently, a gene lying approximately 110 kb telomeric of the bcl-1 major translocation cluster breakpoint locus, designated PRAD1, was proposed as a candidate bcl-1 oncogene. Accumulated evidence now indicates that this gene is the postulated bcl-1 oncogene. It encodes a protein with homology to cyclin family proteins designated cyclin D1 (CCND1). In order to determine whether 11q13 translocation breakpoints were present near the PRAD1 coding region in addition to the previously defined bcl-1 sites, we analyzed 27 centrocytic lymphomas by Southern blot using genomic and cDNA probes flanking the first exon of PRAD1. Five samples showed rearrangement at PRAD1 sites. In four of these, the breakpoints could be mapped from approximately one to 25 kb upstream of the first PRAD1 exon; each showed comigration of rearranged PRAD1 and immunoglobulin heavy chain joining gene bands consistent with the t(11;14)(q13;q32). The fifth case was rearranged with PRAD1 probes only on BamHI-digested DNA, indicating either a point mutation or a polymorphism at this site. This sample also had rearrangement on multiple enzyme digests with the bcl-1 p94PS probe. None of 80 non-centrocytic B-cell neoplasms showed PRAD1 rearrangement. Thus, rearrangement at both bcl-1 and PRAD1 loci is strongly associated with centrocytic lymphoma, and provides a useful molecular marker for classifying this subtype of lymphoma. Furthermore, translocation-induced aberrant expression of the PRAD1 cyclin may lead to deregulated cell cycle control and play an important role in the pathogenesis of centrocytic lymphoma. PMID- 8426478 TI - Selective increase of alternatively spliced Lck transcripts from the proximal promotor in hematopoietic malignancies. AB - Screening by Northern blot for lck expression in 51 patients with diverse hematologic diseases has shown, for four of them, a 3 to 15-fold increase in the level of lck mRNA when compared with expression in healthy donors. These patients suffered from diverse malignancies: one Burkitt lymphoma, one T-cell lymphoma and two non-Hodgkin B-cell lymphoma. Specific analysis of the different lck transcripts in these patients by polymerase chain reaction and their relative quantitation demonstrate a significant increase of only the type IB and the type IC lck transcripts arising from the proximal promotor. Our study shows: (a) a high lck expression may occur in diverse hematologic diseases, (b) whatever the type of malignancy, this high expression results in a specific increase of the spliced transcripts arising only from the proximal promotor, and (c) in these four patients without any rearrangement or amplification, the high lck expression probably results from the specific activation of the proximal promotor. PMID- 8426479 TI - Polymerase chain reaction analysis of allele frequency and loss at the Harvey ras locus in myeloid malignancies. AB - The hypothesis that 'rare' variable number tandem repeat (VNTR) alleles of the Harvey ras (Ha-ras) locus are an inherited predisposing factor in myeloid malignancies has been evaluated. We describe an application of the polymerase chain reaction (PCR) which amplifies the VNTR region at the Ha-ras locus and offers a number of advantages over conventional Southern analysis. Ha-ras VNTR genotypes were assigned to 57 normal subjects, 46 patients with acute myeloid leukaemia (AML), 26 with myelodysplastic syndrome (MDS) and 49 with chronic granulocytic leukaemia (CGL). By comparison with previous reports we found significantly higher frequencies of rare alleles (20.2%) in our normal subjects of whom more than 35% had at least one 'rare' allele. The frequencies of rare alleles in the patient groups was not significantly different from the normal group (chi 2 = 0.54, p = 0.91). In studies of constitutional and leukaemic DNA from patients with AML, we found that allelic loss at the Ha-ras locus was not a common phenomenon. The improved resolution achievable with PCR compared with Southern analysis was demonstrated by the inability of Southern analysis to resolve six out of 34 PCR heterozygotes. We therefore suggest that previous studies showing linkage between rare Ha-ras alleles and susceptibility to malignancy should be reevaluated using our sensitive PCR technique. PMID- 8426480 TI - Hypermethylation of the calcitonin gene in the myelodysplastic syndromes. AB - It is well documented that the calcitonin gene area in the short arm of chromosome 11 is hypermethylated in most acute leukemias as well as in chronic lymphatic leukemia. In contrast, the gene is normally methylated during the chronic phase of the chronic myeloid leukemia but turns hypermethylated as the disease escalates. As the methylation of the calcitonin gene correlates with the disease activity in chronic myeloid leukemia, it seemed worthwhile to study the gene methylation in other premalignant hematologic conditions with a potential to terminate in fulminant acute leukemia. We report here on the calcitonin gene methylation in patients with myelodysplastic syndromes (MDS) using a methylation sensitive restriction enzyme HpaII and standard Southern blotting techniques. Bone marrow aspirates from a total of 26 MDS patients were studied. In 24 of these patients, the calcitonin gene was hypermethylated. There was no correlation between the methylation status and the morphological stage of the disease. All six patients with a blast count < 5% had a hypermethylated gene. Of the 19 patients with a blast count > 5%, 17 were hypermethylated only two having normal methylation status of the gene. It appears that the hypermethylation of the calcitonin gene area in the short arm of chromosome 11 may be an early event in the pathogenesis of the myelodysplastic syndromes. The methylation analysis may thus be of value as a diagnostic tool in MDS but an abnormal methylation pattern does not seem to have a direct relation with the degree of blast infiltration. PMID- 8426481 TI - Bcl-2 gene rearrangements in primary B-cell lymphoma of the gastrointestinal tract reveal follicular lymphoma as a subtype. AB - The principal objective of this study was to investigate whether follicular center cell lymphomas occur among B-cell lymphoma of mucosa-associated lymphoid tissue (MALT). We used a molecular genetic/immunohistochemical approach and analysed 21 cases with the primary site in the gastrointestinal tract. Only two bcl-2 gene rearrangements were detected in our series and were found in two out of seven lymphomas with a nodular growth pattern. A chromosomal translocation t(14;18) was demonstrated by comigration of rearranged bcl-2 and JH sequences in one of these two cases. Additionally, both lymphomas showed bcl-2 protein positive neoplastic follicles, CD10 expression, and lack of vimentin. Therefore, these two cases were defined as follicular lymphomas. In contrast to the two follicular lymphomas of MALT, three other, nodular growing, bcl-2 protein positive lymphomas were found to have no bcl-2 gene rearrangements, to be CD10 negative and to express vimentin. These three lymphomas might be composed of neoplastic extrafollicular cells which secondarily invaded reactive follicles. We conclude that the presence of bcl-2 protein positive follicles is consistent with both a follicular and extrafollicular origin of a B lymphoma of MALT. However, the detection of a bcl-2 gene rearrangement is the most valuable criterion in such a situation, and additional immunophenotypic criteria, such as CD10 expression and lack of vimentin within the neoplastic population, further substantiate the diagnosis of a follicular lymphoma in MALT. PMID- 8426482 TI - Establishment of a CD4-positive plasmacytoma cell line (AMO1). AB - A human plasmacytoma cell line (AMO1) was established. The AMO1 cells had the light and electron microscopic characteristics typical of plasmacytoma cells and did not harbor Epstein-Barr virus. These cells expressed cytoplasmic immunoglobulin A kappa and the immunoglobulin heavy-chain gene (JH) and kappa light-chain gene (C kappa) were rearranged. Coexpression of a CD4 antigen and plasma cell antigens (CD38 and PCA-1) was an unusual and sustained feature. Neither the T-cell receptor beta nor the gamma chain gene displayed the rearranged form. Other lineage-specific surface antigens, namely T, B, monocytoid, and myeloid antigens, were all negative in AMO1. In accordance with the surface CD4 expression, polymerase chain reaction analysis indicated constitutive expression of CD4 mRNA, and the cytogenetic findings revealed that AMO1 cells had a derivative chromosome 12, which had a structural abnormality of the short arm carrying the CD4 gene locus. These findings provide strong evidence for the presence of CD4-positive malignant plasma cells and raise the possibility that the CD4 expression in the AMO1 cell line is closely associated with the derivative chromosome. PMID- 8426483 TI - Development of a lacZ marked WEHI-3B D+ murine leukemic cell line as an in-vivo model of acute non-lymphocytic leukemia. AB - Established leukemic cell lines have been useful models for studying the biology of leukemia. Analysis of the actions of differentiating agents on leukemic cell lines in vivo has been limited by an inability to unambiguously distinguish host hematopoietic elements from differentiated leukemic cells. In order to identify and quantify leukemic cells during in vivo studies, a derivative of the murine myelomonocytic leukemia cell line WEHI-3B D+, which stably expresses beta galactosidase, was constructed utilizing retroviral vector gene transfer. This cell line, termed WEHI-3B D+/lacZ 2.8, demonstrated in vitro growth and differentiation properties similar to the parental cell line. WEHI-3B D+/lacZ 2.8 expressed high levels of beta-galactosidase following prolonged in vitro growth and following differentiation in suspension cultures and clonogenic assays. In vivo, WEHI-3B D+/lacZ 2.8 was leukemogenic and high level expression of beta galactosidase was maintained. Quantification of tissue involvement with WEHI-3B D+/lacZ 2.8 leukemia was performed utilizing staining with the fluorogenic beta galactosidase substrate fluorescein di-beta-galactoside and fluorescence activated cell sorting analysis. In vivo differentiation efficiency following granulocyte colony-stimulating factor (G-CSF) administration was determined using a simultaneous nuclear and cytoplasmic staining procedure. Results indicate that treatment of mice inoculated with WEHI-3B D+/lacZ 2.8 cells with G-CSF administration causes detectable but limited differentiation. PMID- 8426484 TI - Human rIL4 fails to differentiate leukemic B-cell progenitors growing upon S17 stromal cell line. AB - Stromal cells appear to be key regulatory elements in hematopoiesis and lymphopoiesis. Several stromal cell lines can support B lineage by creating a hematopoietic microenvironment via cell contact or regulatory humoral molecules. These activities have been efficiently mediated by an adipocytic stromal cell line 14F1.1 on infant leukemia cells expressing a hybrid pre-B myeloid phenotype. Several murine cell clones, however, are known to have different ability to support growth and/or differentiation of leukemic cells depending on the maturational stage in which malignant cells are frozen. Pre-B-cell lines and fresh leukemias were therefore cultivated on S17 stromal cell line, before and after exposure to human recombinant interleukin 4 (rIL4), a cytokine whose effects on the growth and differentiation of the B-cell compartment depend on the developmental stage of the target B cell. In the present work, leukemic cells, both in suspension and in close contact with stromal cells, maintained their original phenotype throughout the whole period of co-culture with S17, either before or after exposure to human rIL4. PMID- 8426485 TI - Ph-positive chronic myeloid leukemia mimicking essential thrombocythemia and terminating into megakaryoblastic blast crisis: report of two cases with molecular studies. AB - Two patients with chronic myeloid leukemia (CML) presenting with the hematologic features of essential thrombocythemia (ET) are reported. At diagnosis they showed extremely high platelet counts (4985 and 2800 x 10(9)/l, respectively) and moderate leukocytosis (21 and 17 x 10(9)/l, respectively). In both cases, in addition to the Philadelphia chromosome (Ph), a rearrangement within the major breakpoint cluster region on chromosome 22 was demonstrated, with the breakpoint in the 3' extreme. In patient 1 the disease initially responded to radioactive phosphorus and hydroxyurea, but during the evolutive course a progressive increase in the white blood cell counts was noted, reaching values typical of the chronic phase of CML, and the patient eventually died from blast crisis 45 months after diagnosis. In patient 2, although good control of the platelet counts was achieved with hydroxyurea, the disease also evolved into a blast crisis four months after diagnosis. In both cases monoclonal antibodies and electron microscopy studies demonstrated the megakaryocytic nature of the blast cells. The above features are not consistent with the present and similar cases being Ph positive ET. Instead, they should be regarded as a special form of CML characterized by a marked protagonism of the megakaryocytic component. PMID- 8426486 TI - Production of tumor necrosis factor in response to interferon-alpha in hairy cell leukemia. PMID- 8426487 TI - Failure of IL-3 plus IL-6 to induce the proliferation and differentiation of malignant plasma cell precursors from peripheral blood in multiple myeloma. PMID- 8426488 TI - Myelodysplastic syndrome evolving into a myeloproliferative disorder: one disease or two? AB - We report two patients who had been initially diagnosed as having a myelodysplastic syndrome but subsequently progressed into a leukothrombocytosis state which mimicked a chronic myeloproliferative disorder. Both patients had anemia and mild neutropenia without thrombocytopenia at the time of their diagnosis of myelodysplastic syndrome, and dyshematopoietic features were present in the bone marrow. After treatment with vitamin D3 for 7 and 18 months, respectively, they developed leukothrombocytosis which responded to hydroxyurea. We speculate that these and other similar patients with this unusual course might constitute an entity distinct from the typical myelodysplastic syndromes or chronic myeloproliferative disorders. PMID- 8426489 TI - [Asymptomatic hyperuricemia: primary? secondary? when to treat?]. PMID- 8426491 TI - [Clinical pedagogy as a new branch of medical humanism: the aspects of nondirectivity]. PMID- 8426490 TI - [Histiocytic lymphoma and pulmonary fibrosis in a female patient with essential thrombocythemia]. AB - The case of a patient with the diagnosis of essential thrombocythemia is presented. Following treatment with melphalan during three years the patient presented clinical and radiologic data of pulmonary fibrosis. Thoracotomy with lung biopsy histologically proving fibrosis was performed. The patient developed a true histiocytic lymphoma afterwards. The rarity of pulmonary fibrosis induced by melphalan and the exceptional association of essential thrombocythemia and histiocytic lymphoma is emphasized. The characteristics of the latter disease, diagnostic difficulties and possible treatment are commented upon. PMID- 8426492 TI - [Drug-induced hematuria]. PMID- 8426493 TI - [The use of the potential years of life lost between the ages of 1 and 64 as an indicator of premature mortality in Aragon]. PMID- 8426494 TI - [The apheresis of low-density lipoproteins in homozygous familial hypercholesterolemia]. PMID- 8426495 TI - [Acute endocarditis due to Streptococcus pyogenes]. PMID- 8426496 TI - [AIDS in the elderly patient]. PMID- 8426497 TI - [The cost of education]. PMID- 8426498 TI - [Antiribosomal antibodies and the neurological manifestations in systemic lupus erythematosus]. AB - BACKGROUND: The possible positive correlation between the presence of antiribosomic antibodies and neurologic and neuropsychiatric manifestations in patients with systemic lupus erythematosus, cited in the literature, was analyzed. METHODS: Neurological involvement (current or previous) was evaluated in 71 successive patients. The anti-ENA antibodies (extractable nuclear antigens) were studied with special attention to the antiribosomic antibodies by: a) indirect immunofluorescence (IFI) on triple rat substrate, Hep2 and Crithidia luciliae; b) counter immunoelectrophoresis (CIE); c) double immunodiffusion (DI), and d) Western blot (WB) in the Molt-4 cell line. The statistical study was performed by the Fisher exact test. RESULTS: RibosomAL dyeing was observed in only 2 patients by IFI on triple rat substrate and HEp2. With CIE and DI, 14 patients (20%) were anti-ENA positive. Detectable bands were obtained with WB in 47 patients (66%) with 9 corresponding to antiribosomic antibodies. No statistical differences were found (p > 0.3) in relation with the presence of antirobosomic antibodies in particular and anti-ENA in general, between the groups with and without neurologic involvement. CONCLUSIONS: No relation was observed between antiribosomic antibodies and neurolupus by determinations of anti-ENA antibodies by Western blot (superior method--p < 0.0001--to direct immunofluorescence, counter immunoelectrophoresis and double immunodiffusion in the comparative study of sensitivity. PMID- 8426499 TI - [A cost-efficacy analysis of treatment with antiendotoxin monoclonal antibodies in gram-negative sepsis]. AB - BACKGROUND: The aim of this study was to determine the cost-efficacy analysis of treatment with antiendotoxin monoclonal antibodies in adult patients admitted to intensive care units for completing the decision making regulating the authorization of registry of drugs in Spain. METHODS: Two treatment strategies were considered: to treat with antiendotoxin monoclonal antibodies, addition to conventional treatment, to all patients with sepsis or to all those with septic shock versus the alternative of only carrying out conventional treatment. The economic evaluation technique which quantifies costs in pesetas and efficacy in years of life gained by the treatment alternatives was considered. RESULTS: The cost per year of additional life gained from treatment to all patients with sepsis was higher (859, 288 pesetas per year of life gained) than that in patients treated for septic shock (293, 810 pesetas per year of life gained). The result was very sensitive to changes in those expected in survival of the patients treated. CONCLUSIONS: In agreement with this cost efficacy analysis the hypothesis to authorize drug registry in Spain for the indication of septic shock is reinforced. The results may vary upon the obtaining of more clinical and epidemiologic information of the treatment. PMID- 8426500 TI - [The characteristics of digoxin utilization in a population under emergency care for heart failure or auricular fibrillation]. AB - BACKGROUND: With the aim of studying the characteristics of the use of digoxin in a population which attended a hospital emergency department for heart failure or auricular fibrillation a 3 month observational study was carried out in the emergency department of the Ciudad Sanitaria de la Vall d'Hebron. METHODS: One hundred twelve patients treated with digoxin who went to the emergency department for heart failure and/or decompensated auricular fibrillation were studied. Clinical and pharmacological histories, determination of digoxinemia and the usual complementary explorations were performed. RESULTS: It was found that 50% of the patients were not adequately controlled and treatment was not followed in 21% of the patients. No significant relation was found between the doses of digoxin and the age of the patients. In multivariant analysis (multiple lineal regression) digoxinemia was related with the doses (beta = 0.22, p = 0.01), cardiac frequency (beta = 0.19, p < 0.05), and compliance (beta = 0.18, p = 0.05). Among the patients in whom the cause of decompensation of cardiac failure could not be identified, one third (31%) were found to have infratherapeutic digoxinemia. CONCLUSIONS: Most patients with cardiac decompensation attending an emergency department are those who are not adequately controlled in primary health care and the rate of incomplete following of the prescription is high. Furthermore, one third of the patients who decompensate with no clinically apparent reason has an infratherapeutic plasma concentration of digoxin. PMID- 8426501 TI - [Autoantibodies in systemic lupus erythematosus]. PMID- 8426502 TI - Laryngeal reconstruction following vertical partial laryngectomy. AB - Laryngeal reconstruction following vertical partial laryngectomy (VPL) with arytenoidectomy was studied in 30 dogs. Variations of a superiorly based thyroid cartilage flap (TCF) were used for reconstruction. In addition, sternohyoid muscle reconstruction and endolaryngeal muscle coverage were studied. Larynges were recovered after a 6-month period in 28 surviving dogs and were analyzed by endoscopic photographs and axial whole-organ sections. Endoscopic assessment (n = 18) demonstrated good results for arytenoid replacement (100%), pseudocord position (94%), pseudocord development (94%), and airway patency (100%). Arytenoid replacement was judged as completely (78%) or partially (22%) replaced. This was accomplished by a pseudocord extending to the cricoid in the horizontal plane. Pseudocord position was judged as normal (83%) or paramedian (11%), with the remainder lateralized (6%). Pseudocord development was judged as complete (72%) or partial (22%), with the remainder poor (6%). Reviewing both endoscopic photographs and gross sections (n = 28), airways were all normal without laryngeal or tracheal stenosis. Histologic assessment (n = 24) also demonstrated good results for arytenoid replacement (79%), pseudocord position (87%), and TCF survival in the glottic plane (79%). Arytenoid replacement was judged as complete (62%) or partial (17%), with the remainder poor (21%). Pseudocord position was judged as normal (50%) or paramedian (37%), with the remainder lateralized (13%). TCF survival was judged as total (63%) or partial (16%). Although not present in the glottic plane in the remaining cases (21%), a portion of the TCF was always present in the supraglottic region. The TCF was largely replaced by bone in the region of the pseudocord, and was covered by nonkeratinizing stratified squamous epithelium and a thick fibrous layer. Breakdown over the TCF was infrequent, with a small focus of granulation tissue over cartilage present in 1 (4%) of 24 cases. Clinically insignificant granulation tissue was present in a total of 6 (25%) cases. In the other 5 cases, this was over muscle or over permanent sutures. Focal cartilage necrosis was present in 2 (8%) of 24 cases, and was localized, self limiting, and deep to the endolaryngeal surface. When the TCF failed to survive histologically, poorer results for arytenoid replacement and pseudocord position generally resulted. However, this apparent difference was not statistically significant due to small sample sizes and variability in results. Other factors that may have kept this difference from becoming larger were thought to be contraction of th e normal cord towards the operated side with fore shortening of the glottis, and medial rotation and ossification of the posterior thyroid ala remnant.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8426503 TI - The clinical differentiation between vocal cord paralysis and vocal cord fixation using electromyography. AB - With newer techniques for laryngeal intervention, it becomes a practical necessity to understand whether an immobile cord is due to neurogenic dysfunction or cricoarytenoid fixation. An objective test for this differentiation is laryngeal electromyography, which can be done as an office procedure with a minimum of discomfort. Our experience in a clinical setting has shown laryngeal electromyography to be efficient in accurately assessing the neuromuscular status of the intrinsic laryngeal musculature. PMID- 8426504 TI - Effect of cricopharyngeus myotomy on postlaryngectomy pharyngeal contraction pressures. AB - After total laryngectomy, the cricopharyngeus muscle, when intact, appears to inhibit the free flow of saliva and secretions past the pharyngeal repair into the upper esophagus. The authors hypothesize that cricopharyngeus myotomy reduces sphincteric pressure, thereby diminishing forces against the pharyngeal suture line. Peak pharyngeal pressures were recorded in patients who underwent total laryngectomy with and without cricopharyngeus myotomy. In patients without concurrent myotomy, peak pharyngeal pressures were all greater than 60 mm Hg. With concurrent myotomy, peak pharyngeal pressures averaged less than 40 mm Hg. Concurrent myotomy carries with it the potential for minimizing postoperative fistulization, eliminating dysphagia of cricopharyngeus spasm, and improving the acquisition of alaryngeal speech. PMID- 8426505 TI - Nonpalpable occult and metastatic papillary thyroid carcinoma. AB - The results of palpation, ultrasound, and detailed pathology were compared in 50 patients undergoing total thyroidectomy. Of the 211 nodules recognized by pathology, palpation detected 24% and ultrasound detected 43%. There were 14 patients with palpable papillary carcinomas, and 12 of these 14 had metastatic foci in other portions of the gland. Metastatic foci would have been left in 10 patients if only a lobectomy had been performed and would have been left in 6 patients if only a subtotal thyroidectomy (as defined in this report) had been performed. Of the 33 patients with benign palpable nodules, 5 had occult papillary carcinomas. In 2 of those 5 patients, the occult carcinomas would not have been removed if a less-than-total thyroidectomy had been performed. The significance of these occult and metastatic papillary carcinomas is discussed. PMID- 8426506 TI - Easy bruisability, aspirin intolerance, and response to DDAVP. AB - Easy bruisability raises the issue of bleeding during otolaryngological surgery. Ten female patients with easy bruisability were evaluated by aspirin challenge; clinical history and screening coagulation studies in these patients had revealed no evidence of a bleeding disorder. The baseline Ivy bleeding time (BT) test (4.5 to 9.5 minutes) was found to be normal in 6 patients and prolonged in 4 patients. Following treatment with aspirin, the bleeding time prolonged significantly in the three groups evaluated: normal controls (6.0 +/- 1.5 minutes vs. 8.4 +/- 2.0 minutes), patients with easy bruisability and a normal baseline (7.8 +/- 1.3 minutes vs. 12.0 +/- 1.6 minutes), and patients with easy bruisability and an abnormal baseline (11.0 +/- 0.7 minutes vs. 14.5 +/- 0.9 minutes). Administration of DDAVP (desmopressin acetate) 0.3 microgram/kg normalized the prolonged bleeding times in all groups after 7 days of daily aspirin therapy. Performing bleeding times before aspirin challenge, after aspirin challenge, and after DDAVP therapy following aspirin challenge is both a useful way of confirming aspirin sensitivity in patients with easy bruisability as well as a useful way of documenting improved hemostasis after DDAVP administration. PMID- 8426507 TI - Congenital perilymphatic fistula and associated middle ear abnormalities. AB - To specifically determine the frequency and type of middle ear abnormalities associated with perilymphatic fistula (PLF), a retrospective chart review was performed of 94 patients (117 ears) who underwent exploratory tympanotomy for PLF from 1980 to 1989. Of the 117 ears explored, 80 (68.4%) had a PLF, and in 65 (81.3%) of these ears, a middle ear malformation was associated with the PLF. Of these 65 ears in which a congenital middle ear abnormality was observed, a malformed stapes was the most common abnormality seen (39 ears, 60%), followed by a deformed round window (20 ears, 30.8%), a deformed incus (11 ears, 16.9%), and a deformed promontory (2 ears, 3%). Often these malformations coexisted amongst themselves or with inner ear abnormalities. Sixteen children (25 ears) had an inner ear malformation identified on computed tomography (CT); all of these children had a PLF found at the time of surgery. This study demonstrated that 86.3% of the ears found to have a PLF had a deformity of the middle ear, inner ear, or both. A malformation of the stapes, most frequently identified as a deformity of its superstructure (and presumably also the anterior footplate), was the most common congenital middle ear abnormality found to be associated with PLF in children. PMID- 8426508 TI - The three bellies of the canine posterior cricoarytenoid muscle: implications for understanding laryngeal function. AB - The posterior cricoarytenoid (PCA) muscle is known to be active during phonation and respiration. The presence of muscle compartments (bellies) that might subserve these functions was investigated in the canine PCA by anatomical dissection and muscle fiber histochemistry. Five PCA muscles were microdissected and the origins and insertions of all muscle bundles were recorded. An additional six PCA muscles were frozen, sectioned, and stained for adenosine triphosphatase (ATPase) activity. The total number of fast- and slow-twitch fibers were counted and their proportion was determined for each region of the muscle. The PCA muscle was found to contain three distinct neuromuscular compartments. The vertical compartment is oriented at 24 degrees from true vertical, inserts on the lateral aspect of the muscular process of the arytenoid, and is composed of 65% type 2 (fast) muscle fibers. The oblique is oriented at 44 degrees from vertical, inserts on the top of the muscular process of the arytenoid, and is composed of 77% type 2 muscle fibers. The horizontal is oriented at 63 degrees from vertical, inserts on the medial aspect of the muscular process of the arytenoid, and is composed of 59% type 2 muscle fibers. The cricoarytenoid joint is capable of three arcs of motion and the physical arrangement of each compartment appears to correspond to each of these motions. Moreover, the histochemical profiles show that the activity of the three bellies is quite different. These results suggest that the different compartments of the PCA perform distinctive motions during phonation and inspiration. PMID- 8426509 TI - Histologic correlation of the degenerating facial nerve with electroneurography. AB - The myogenic compound action potential (CAP) measured by electroneurography (EnoG) is hypothesized to correlate directly with the number of viable motoneurons in the facial nerve. In an animal model (cat), two independent ENoG techniques, standardized recording lead placement (SRLP) and optimized recording lead placement (ORLP), were used to record CAPs evoked from nerves undergoing degeneration. Normal test-retest variability simulated human studies. Peripheral counts of viable axons correlated with N1 amplitude (ORLP ENoG), peak-to-peak (absolute) amplitude (both ENoG techniques), and area under the negative phase of the diphasic CAP waveform (both ENoG techniques) recorded over the whisker pad. These results validate ENoG as a reliable indicator of neural integrity following traumatic lesions of the facial nerve. PMID- 8426510 TI - Pharmacologic manipulation of random skin flaps with pentoxifylline. AB - The literature demonstrates inconsistent results amongst investigators who have used pentoxifylline in an effort to enhance skin flap survival. This study employed a standardized skin flap model in the rat and a standard intraperitoneal dose of pentoxifylline (10 mg/kg) delivered in four different temporal regimens. Fluorescein staining and length of flap survival were measured. The only regimen that demonstrated increased flap survival over saline controls included administration of the drug immediately upon raising the flap and every 12 hours for the next 7 days. Three different regimens that included preoperative administration of the drug failed to demonstrate a beneficial effect on skin flap survival. Slight improvement in flap survival was seen in animals receiving fluorescein. These results suggest an inconsistent pentoxifylline effect even within a single controlled study. PMID- 8426511 TI - Eosinophil cationic protein in the nasal secretions of patients with mite allergic rhinitis. AB - In 25 patients with mite nasal allergy and 13 healthy control subjects, levels of eosinophil cationic protein (ECP) and histamine in nasal secretions were examined before and after challenge with mite extract. The ECP level was found to be significantly higher in the washings taken 30 minutes after challenge compared with the washings taken before challenge (P < .05). ECP levels measured both before and after mite extract challenge were found to be significantly higher in the patients than in the controls (P < .01). These facts suggest that the ECP affects the nasal mucosa by causing injury to ciliated cells and by acceleration of the processes of modification and inhibition of the allergic reaction. PMID- 8426512 TI - Biochemical aspects of autologous fibrin glue derived from ammonium sulfate precipitation. AB - The autologous fibrin glue (AFG) was produced from 30 human donors using the ammonium sulfate precipitation method, and a study of its biochemical composition was performed. The fibrinogen concentration ranged 13 to 57 mg/mL, the yield averaged 54.6%, and there was a direct relationship between the level of fibrinogen in plasma and AFG which had been made from the same donor's plasma. Our results suggest that the quality of this category of AFG depends partially on the fibrinogen level of donor's plasma. Quality control of the AFG is discussed. Additionally, the effectiveness of three fibrinolysis inhibitors were compared to find an optimum one which inhibits autofibrinolysis of this glue. Tranexamic acid and epsilon aminocaproic acid (EACA) were acceptable alternatives to aprotinin. PMID- 8426513 TI - Fibrin binding, fibrinolytic and fibrinogenolytic activity of plasminogen activator derived from the paranasal mucous membrane of humans. AB - It is known that large amounts of plasminogen activator (PA) are contained in tissue extracts of the human paranasal mucous membrane (PMM) with chronic sinusitis. The present study was undertaken to isolate and purify the PA in tissue extracts of PMM. Furthermore, the purified PA was identified as to whether it was of the tissue type or urokinase (UK) type, and some of its fibrinolytic characteristics were determined in comparison with those of urokinase. As starting material, extracts of acetone powder of PMM with chronic sinusitis were used, and Zn-imminodiacetate affinity chromatography, and ultrafiltration were carried out to separate and purify the PA from the PMM. The PA was purified to a 107-fold increase in specific activity. The molecular weight of the PA was estimated to be 65,000 to 70,000 d by gel filtration using Sephacryl S-200. The purified PA was stable in the range of pH 8.0 to 9.0. Using S-2288, a synthetic substrate, the Michaelis constant (Km) of the purified PA was estimated to be 0.11 mmol. The binding of the purified PA to fibrin was stronger than that of UK, while the fibrinogenolytic activity of the purified PA was not stronger than that of UK. Based on these results, the purified PA was identified as a tissue-type plasminogen activator (t-PA). From the kinetic data, it was identified as being of the two-chain variety. It is considered that, as a thrombolytic agent, t-PA derived from the PMM could be more useful than UK. PMID- 8426514 TI - Treatment of the compromised trachea with sleeve resection and primary repair. PMID- 8426515 TI - Double-barreled (diversionary) tracheostomy: long-term results and reversibility. PMID- 8426516 TI - Adipose myringoplasty in children. PMID- 8426517 TI - Preliminary analysis of histological results of Hexascan device with continuous tunable dye laser at 514 (argon) and 577 NM (yellow). AB - This histological study demonstrates that the Hexascan device combined with a continuous tunable dye laser at either 514 nm or 577 nm is able to effectively treat port wine hemangiomas. Both 514 nm and 577 nm were effective in producing almost normal looking skin histologically after treatment in some patients. Other patients demonstrated a minor number of residual vessels or no significant change. In this small series, it appeared that the 514 nm and 577 nm wavelengths were similar histologically. PMID- 8426518 TI - Rationale for automatic scanners in laser treatment of port wine stains. AB - Laser therapy of port wine stains (PWS) is well established. However, the application of lasers to PWS treatment over the last decade has resulted in the use of many laser systems. The goal of this study is to describe the different laser parameters, treatment techniques, and delivery systems. Two methods, coagulation and microvaporization, are used to destroy the abnormal blood vessels. The coagulation method results in excellent treatment, but it requires careful control of laser parameters during the treatment process. Scanning devices can be utilized for this purpose. The microvaporization method, obtained with a pulsed dye laser, is restricted to PWS composed of small vessels since it is difficult to obtain a pulse longer than 450 microseconds. In addition, numerous retreatments are needed with the pulsed dye method. The initial assessment of the patient should contribute to an improvement in the choice of laser parameters and the outcome of treatment. PMID- 8426519 TI - Absorption characteristics at 1.9 microns: effect on vascular welding. AB - A 1.9 microns laser was used to investigate the acute weld strengths for anastomoses of rat and rabbit aortas and femoral arteries. The wall thicknesses for these vessels approximately matched the optical absorption depth of 125 microns for 1.9 microns radiation in vascular tissues. A low power (150 mW) 1.9 microns laser was used. Laser power was delivered through silica fiber optics for manual control. The fiber tip was held approximately 1 mm from the target resulting in a laser spot size of 0.7 mm at the tissue. The linear delivery rate was approximately 0.3 mm/sec. Acute burst pressures of the welds showed a linear correlation with the reciprocal of the vessel radius. These results suggest that the product of the weld strength times the optical absorption depth is constant over the range of vessel sizes studied. A weld strength for a weld thickness equal to the optical absorption depth was determined to be 4 x 10(6) dynes/cm2, which is comparable to the strength of sutured anastomoses. These acute studies suggest that a laser wavelength with absorption depth in tissue matched to the vessel wall thickness should yield optimum welds. Therefore, a laser operating near 1.9 microns is suitable for small vessel welding. PMID- 8426520 TI - Relation between skin surface temperature and minimal blanching during argon, Nd YAG 532, and CW dye 585 laser therapy of port-wine stains. AB - Laser photocoagulation has proven to be valuable in the treatment of port-wine stains. In this application, the minimal blanching technique is used as an indicator of suitable dosage since it has been demonstrated that the immediate appearance a white mark is required to achieve permanent blanching a few months later. The objective of the investigations undertaken in this study was to correlate the temperature attained at the surface of port-wine stains with immediate blanching, upon irradiation with different laser fluences. A comparative study was performed using an argon laser (all lines), a 532 nm Nd:YAG and a 585 nm argon pumped dye laser. Surface temperature was studied using an infrared camera. Temperature was measured on 10 different port-wine stains using different fluences. Whitening threshold fluence was related to surface temperature. It appeared that whitening threshold fluence corresponded to a surface temperature of 53 degrees C (+/- 3 degrees C). The whitening threshold fluence was dependent on port-wine stains and wavelength. However, whitening threshold fluence remained lower for 532 nm and 585 nm and it correlated to the absorption curve of hemoglobin. PMID- 8426521 TI - Pulsed dye laser treatment of recalcitrant verrucae: a preliminary report. AB - Patients with recalcitrant warts on the fingers and hands, periungual, and other parts of the body including verrucae plana and plantar surfaces were treated using the pulsed dye laser at 585 nm, 450 usec, and a spotsize of 5 mm diameter. Of the 39 patients treated, 28 (72%) were cleared of their warts after an average of 1.68 treatments at fluences of 6.25-7.5 J/cm2. Seven (18%) patients had a reduction of between 80-95% of their warts after 1.3 treatments, and verrucae reduced by 50% in four of the 39 patients after one treatment. The average follow up period of the 28 cases cleared of their warts has been 5 months. Of this group, those with periungual warts have been followed for up to 6.4 months, compared to 4.8 months for those with warts on other parts of their body, 4.0 months for those with finger and hand warts, and 2.0 months for plantar warts. Only one of the 28 patients has had a recurrence after 3 months of clearance. PMID- 8426522 TI - Viable bacteriophage in CO2 laser plume: aerodynamic size distribution. AB - The size of CO2 laser generated plume particles containing viable bacteriophage, phi X174, was determined with 2 models (10-830 and 15-500) of Andersen bioaerosol cascade impactors. Samples were collected during 100 one-second laser exposures (approximately 68 W/cm2) of a bacteriophage-agar substrate with and without a space-confining hood. The hood appeared to facilitate collection of the largest particles (P < 0.1). In addition, Andersen model 15-500 was a more efficient collector of the largest particles, a phenomenon which is likely a function of the dynamic nature of the laser plume as well as impactor design. We found that laser plume particles containing viable bacteriophage are very large, in one instance exhibiting a count median aerodynamic diameter (CMAD) and mass median aerodynamic diameter (MMAD) of 23 and 55 microns, respectively. Furthermore, the appearance of viable bacteriophage in these plume particles constitutes an extremely rare event. The limitations of cascade impactor design when used to analyze particles with high water concentrations generated in a laser plume are discussed. PMID- 8426523 TI - Quantification of phthalocyanine concentration in rat tissue using laser-induced fluorescence spectroscopy. AB - Quantification of photosensitizer concentration in tissue should improve planning and outcome of photodynamic therapy. Laser-induced fluorescence (LIF) can be used to measure in vivo fluorescence of photosensitizers in tissue. This study was designed to determine if in vivo fluorescence intensity of chloroaluminum phthalocyanine tetrasulfonate correlates with its concentration in different rat tissues. Following LIF measurements, the animals were humanely euthanized and the concentration of phthalocyanine in different tissues was determined using chemical extraction technique. The correlation of phthalocyanine fluorescence intensity and its concentration was determined for each tissue using Pearson product-moment correlation analysis. A strong correlation between in vivo phthalocyanine fluorescence intensity and its concentration was found for spleen, kidney, liver, and chemically induced mammary adenocarcinoma. Low correlation was found for thigh skin and planum of nose. No correlation was found for thigh muscle and tongue. PMID- 8426524 TI - Optical properties of rat liver and tumor at 633 nm and 1064 nm: photofrin enhances scattering. AB - Predicting the effects of photodynamic therapy (PDT) and Nd: YAG laser coagulation requires knowledge of the light distribution in tumor and surrounding tissue. Therefore, absorption and scattering coefficients and the average cosine of the scattering angle (the anisotropy factor) were measured in rat liver and tumor at 632.8 and 1064 nm. A syngenic colon adenocarcinoma CC531 was implanted subcutaneously in two groups of 7 Wag/Rij rats. In one group Photofrin was administered 5 mg/kg intravenously 48 h before determination of optical properties. Two months after inoculation, samples were taken from tumor and liver and optical properties determined using indirect methods with two integrating spheres. The absorption coefficient was larger in liver than in tumor at 632.8 nm (P < 0.0005), whereas the scattering coefficient was larger in tumor than in liver at 1,064 nm (P < 0.05). Addition of Photofrin increased the scattering coefficient in liver and in tumor at both wavelengths (P < 0.025) and decreased the anisotropy in tumor (P < 0.025), suggesting that for modelling the dosimetry of PDT the optical properties of photosensitized tissue should be used. PMID- 8426525 TI - Photoinhibition of smooth muscle cell migration: potential therapy for restenosis. AB - Evidence from animal, autopsy, and atherectomy studies demonstrates that migration and proliferation of smooth muscle cells of medial origin result in neointima formation and decreased luminal cross-sectional area. The purpose of this study was to evaluate whether low energy light irradiation can inhibit smooth muscle cell migration and therefore potentially reduce the degree of neointima formation and the incidence of restenosis. The migration kinetics of bovine aortic smooth muscle cell monolayers were examined using a fence assay. The effect on smooth muscle cell migration of irradiation with monochromatic light at wave-lengths ranging from 400 to 700 nm was compared to the migration of cells irradiated with broadband white light or maintained in the dark. Wavelength specific photoinhibition of smooth muscle cell migration was observed; 594-600 nm light reproducibly inhibited migration by 12-29% (P < 0.05). Migration rate was significantly reduced following daily radiant exposures of 1.0 J/cm2 as well as following a single radiant exposure of 0.09 or 0.9 J/cm2. The decrease in migration was not associated with any change in cell proliferation or [3H] thymidine incorporation. We conclude that 594-600 nm light inhibits smooth muscle cell migration in vitro and may potentially be used in vivo to decrease fibrointimal thickening following arterial injury. This application of photoinhibition may be useful in retarding restenosis following angioplasty. PMID- 8426526 TI - Absence of rhodamine 123-photochemotoxicity in human tumor xenografts. AB - Rhodamine 123 (R123)-photochemotoxicity was assessed in BALB/c nude mice bearing a xenografted human squamous cell carcinoma at various power densities and wavelengths and a given incident fluence of 150 Joules/cm2. One hour before light delivery, 1 mg R123/kg was injected i.p. in 20 animals. Surface irradiance was performed on the tumor and an equal size hind leg area of 40 animals. Three groups of 10 animals were treated at 514.5 nm and 0.1 W/cm2, 1 W/cm2, and 30 W/cm2, and one at 488 nm and 30 W/cm2. In each group, five animals received R123. The R123 concentration was measured in the tumor (0.023 +/- 0.007 micrograms/g) and tumor-free tissue (0.023 +/- 0.008 micrograms/g) in three additional animals by high performance liquid chromatography 1 hour after R123-administration. Histologic assessment 72 hours after light delivery revealed no tissue damage at nonthermal power densities, either in the tumor or in the tumor-free tissue, irrespective of R123-administration. At 30 W/cm2, neither in the tumor nor in tumor-free tissues was there any significant difference in the depth of necrosis, irrespective of R123-administration and the wavelength applied. Our results question the validity of R123 as a photosensitizer, at least in this rodent tumor model. PMID- 8426527 TI - Comparison of magnetic resonance images and the histopathological findings of lesions induced by interstitial laser photocoagulation in the brain. AB - Interest has developed in using magnetic resonance (MR) imaging to monitor the volume of tissue destroyed by interstitial laser photocoagulation (ILP). In these experiments, ILP was induced in the normal brains of 9 anesthetized cats by delivering 1.5 W of continuous-wave Nd:YAG laser energy (1,064 nm) from a single 400-microns core optical fiber for 1,000 s. The irradiations were monitored using proton spin-echo MR imaging during and immediately after ILP and at postirradiation survival times of 2, 5, and 14 days. At 2 days postirradiation, the necrotic thermal lesion consisted of a central cavity surrounded by 2 concentric zones of coagulative necrosis, one dense and the other dispersed. The lesion shrank and the zonal appearance became less obvious over the 14 day survival period. An enhancing halo on contrast-enhanced T1-weighted images acquired immediately postirradiation best approximated the total lesion diameter at 2 days. These images also indicated that the volume of tissue destroyed during ILP corresponded better to the necrotic volume determined at 2 days than at 5 days and 14 days postirradiation. T2-weighted images acquired during and immediately after ILP consistently underestimated the total lesion diameter at 2 days. PMID- 8426528 TI - Laser induced wounds and scarring modified by antiinflammatory drugs: a murine model. AB - Scarring is a well-known side effect to cutaneous laser treatment. Therefore we investigated if wounds and scarring could be decreased by pharmacological interference in the acute inflammatory reaction following laser therapy. A copper vapor laser operating at 578 nm was used at three different intensities and doses. The antiinflammatory drugs methylprednisolone (1 mg/kg/24 h) and indomethacin (2 mg/kg/24 h) were administered through the drinking water. The laser induced wounds were evaluated daily and histologic and biochemical analyses were used to estimate scarring. The administration of antiinflammatory drugs resulted at some laser intensities in increased scores of maximum wound area, healing time, cumulated wound area, fibrosis, hydroxyproline, and hydroxylysine, being mostly pronounced for indomethacin at 0.8 W/spot. The maximum wound area and the wound healing time could be used as a predictor of scarring. A dose response was shown between the laser intensities and the mentioned parameters. We recommend carefulness in the laser treatment of patients receiving indomethacin and disrecommend use of the drug in the prophylaxis against laser induced scarring. PMID- 8426529 TI - Sub ablation effects of the KTP laser on wound healing. AB - The KTP laser (wavelength 532 nm) was used in a sub ablative format to determine the effect of low energy density irradiation on the normal healing by primary intention of scalpel skin incisions in rats. Two longitudinal lased strips were created by a 1 cm diameter defocused beam on the shaved, cleaned dorsal epidermis of 32 Sprague-Dawley rates; one strip was produced with a 2.0 W beam (54 J, or 18 J/cm2 total dose), and the other with a 3.5 W beam (94.5 J or 31.5 J/cm2, total dose). Scalpel incisions were made longitudinally within the irradiated zones, using contra lateral scalpel incisions on unirradiated skin as controls. Tensiometric analysis of wound strength was performed at 3, 7, 14, and 23 days following surgery. The data from fresh tissue tensiometry indicate that KTP laser irradiation of skin incisions results in a lower tensile strength for the wound at 7 and 14 days. The decrease in tensile strength is proportional to the total energy density of the exposure. At day 3 and 23, the tensile strength of the wound was independent of the sub ablative laser exposure. The results are in general agreement with studies of the healing process of laser incisions and may help us to understand the details of the healing process from laser incisions. PMID- 8426530 TI - Low power laser irradiation alters the rate of regeneration of the rat facial nerve. AB - Low power laser irradiation has been reported to cause biological effects due to the photochemical and/or photophysical action of the radiation. This study determined quantitatively if transcutaneous low power laser irradiation can affect the regeneration of the rat facial nerve. The facial nerve was crushed unilaterally in anesthetized rats and transcutaneously irradiated daily with a laser beam directed at the area of the crush injury. Laser treatment began on the day of the crush injury and was continued daily for 7, 8, or 9 days. Preliminary experiments determined the most effective wavelength, laser power, length of irradiation, and treatment schedule. The wavelengths examined were 361, 457, 514, 633, 720, and 1064. The laser powers and lengths of irradiation examined ranged from 8.5 to 40 mW and 13 to 120 min. Irradiation treatment was done daily, on alternating days and on the first 4 days postcrush. The most effective laser parameters for the low power treatment included daily irradiation with a helium neon (HeNe) or argon pumped tunable dye laser a wavelength of 633 nm, with a power of 8.5 mW for 90 minutes (45.9 J, 162.4 J/cm2). The number of horseradish peroxidase (HRP) labeled neurons in the facial motor nucleus was used as an assay of the degree of regeneration. In rats in which the facial nerve was crushed but not irradiated, the average number of HRP labeled neurons in the facial nucleus was 22 on day 7 postcrush, 54 on day 8, 116 on day 9, and 1,149 on day 10. After HeNe or argon pumped tunable dye laser irradiation, the average number of HRP labeled neurons increased to 34 on day 7 postcrush, 148 on day 8, and 1,725 on day 9. There was a statistically significant difference between the control and irradiated rats on day 9 postcrush (P < 0.01). These data indicate that transcutaneous low power irradiation with the lasers and parameters involved in this study increased the rate of regeneration of rat facial nerve following crush injury. PMID- 8426531 TI - Evaluation of thermal cooling mechanisms for laser application to teeth. AB - Experimental cooling methods for the prevention of thermal damage to dental pulp during laser application to teeth were compared to conventional treatment in vitro. Pulp temperature measurements were made via electrical thermistors implanted within the pulp chambers of extracted human third molar teeth. Experimental treatments consisted of lasing without cooling, lasing with cooling, laser pulsing, and high-speed dental rotary drilling. Comparisons of pulp temperature elevation measurements for each group demonstrated that cooling by an air and water spray during lasing significantly reduced heat transfer to dental pulp. Laser exposures followed by an air and water spray resulted in pulp temperature changes comparable to conventional treatment by drilling. Cooling by an air water spray with evacuation appears to be an effective method for the prevention of thermal damage to vital teeth following laser exposure. PMID- 8426533 TI - Effects of ArF:excimer laser irradiation on human enamel and dentin. AB - Round enamel and dentin surfaces of sound and carious extracted human teeth were irradiated by an ArF:excimer laser for up to 180 sec. Thermographic measurements indicated that the temperature rise due to heat accumulation caused by laser irradiation on these enamel and dentin surfaces was up to 19 degrees C (10 HZ with 540 J/cm2), and the temperature returned to the preirradiation value within 10 sec after the irradiation was stopped. Under light microscopy, no carbonization was evident on these surfaces, and a simple recess was formed by abrasion or vaporization in the irradiated regions. In the secondary SEM, uniformly distributed fine pores and prism structures appeared slightly on the enamel surfaces. Between the peritubular and the intertubular dentin, there appeared a distinct difference in the dissolved area. The laser almost completely removed carious regions of the enamel and the dentin, and penetration extended beyond the carious regions. In the backscattered electron SEM, highly mineralized layers were observed on the enamel and dentin surfaces dissolved by the laser. PMID- 8426532 TI - Structural changes in dental enamel induced by high energy continuous wave carbon dioxide laser. AB - The morphological changes in dental enamel exposed to a high energy continuous beam surgical carbon dioxide laser were studied with a wet-scanning electron microscope. Utilizing a 1 mm focused beam, energy levels of 5, 10, and 12.5 watts were applied to the external enamel surface of teeth for between 0.05 and 2 seconds. Subsequent examination showed melting and resolidification of the surface enamel. Examination in cross section revealed three different zones of change, which were consistent in thickness. The outer layer of melted and resolidified enamel approximates 100 microns in width, whereas the next zone of acid resistant amorphous enamel measures only 12 microns in width and the third zone of porous enamel measures approximately 110 microns in width. Though of no immediate practical value in dentistry, the exact physical chemical changes occurring in the enamel warrant further investigation. PMID- 8426534 TI - Group B streptococcal meningitis in adults. AB - Group B streptococcal (GBS) meningitis is a frequent entity in neonates but an uncommon cause of meningitis in adults. Retrospective analysis at our institution identified 4 adult cases over the last 25 years; an additional 46 cases from the literature were reviewed. A bimodal age distribution paralleling that seen in other severe GBS infections was observed. Clinical presentation was not unlike meningitis due to other pyogenic organisms, although a higher percentage of patients presented with less than 24 hours of symptoms. Forty-three percent of patients had no underlying illnesses. Concomitant bacteremia was present in 83% of patients. The overall mortality was 27% and was limited exclusively to patients with co-morbid illnesses. Meningitis in adults due to GBS should be considered in the immunocompetent as well as the immunocompromised host. PMID- 8426535 TI - Mycobacterium bovis infections in San Diego: a clinicoepidemiologic study of 73 patients and a historical review of a forgotten pathogen. AB - We have presented 73 patients (48 adults and 25 children) with microbiologically documented M. bovis infections identified over the 12-year period from 1980 through 1991. Epidemiologic investigation of these patients revealed that the majority (80%) were of Hispanic origin. The non-Hispanic patients either had traveled extensively outside the United States, were born in the United States during its endemic period or in other countries with endemic bovine tuberculosis, or were exposed to a close relative with a positive PPD and known exposure to M. bovis. For Hispanic patients, the presence of reactivation disease in adults and primary disease in children indicate that this mycobacterium remains endemic in Mexican beef and dairy herds, a position supported by United States monitoring of Mexican cattle transferred across the border. Our review of the historical and contemporary efforts to eradicate this animal and human pathogen from the livestock industry in the United States and abroad shows that the implementation of similar methods could be effective in Mexico. The detailed presentations of selected patients and summaries of the clinical manifestations in the remainder of our 73 patients reveal striking similarities to historical accounts and to more contemporary studies of reactivated disease in England. Although M. bovis infections are still expressed predominantly in extrapulmonary sites (cervical and mesenteric nodes, the peritoneum, and the GU tract), as many as 50% of adult patients will present only with pulmonary disease. Underlying immunosuppressive disorders were particularly prominent in adults with extrapulmonary disease. For example, HIV positive patients accounted for 12 of 48 adults and 1 adolescent patient in our series. Overall, M. bovis infections accounted for almost 3% of all tuberculous disease reported in San Diego County during the study period. The intrinsic resistance of M. bovis to PZA could threaten the response of patients with bovine tuberculosis to the short-course chemotherapeutic regimens now recommended by the CDC and the American Thoracic Society. We strongly recommend continued surveillance for this forgotten pathogen because the importation of Mexican cattle, the migration of Hispanic immigrants from border areas to the United States interior, and the persistence of extrapulmonary disease in immunocompetent and HIV-infected United States citizens assure its persistence in this country. PMID- 8426536 TI - Bullous amyloidosis. Report of 3 cases and review of the literature. AB - We report 3 cases of bullous immunoglobulinic amyloidosis and review 25 published cases. In 2 of our patients, amyloid deposits were not detected with special staining, but by means of ultrastructural methods. Investigations of the skin lesions permitted the diagnosis of associated plasma cell dyscrasia in 2 patients. Unexplained bullous lesions should be investigated for amyloid deposits and the presence of monoclonal gammopathy by methods including electron microscopy and immunochemical analysis of serum and urine. PMID- 8426537 TI - Primary systemic amyloidosis: a review and an experimental, genetic, and clinical study of 29 cases with particular emphasis on the familial form. 1956. PMID- 8426539 TI - Night-time curfew: an option for reducing probationary driver casualties. PMID- 8426538 TI - Reducing teenage access to cigarettes in Australia: time to act? PMID- 8426540 TI - The ethics of using animals in biomedical research. PMID- 8426541 TI - Oncology curricula in Australia. PMID- 8426542 TI - The reform of medical education. PMID- 8426543 TI - Tobacco and alcohol use among Australian secondary school students in 1990. AB - OBJECTIVE: To obtain up-to-date prevalence estimates of tobacco smoking and alcohol drinking among Australian secondary students, and to compare these estimates with those obtained from similar studies conducted in 1984 and 1987. DESIGN: Data were collected from 24,892 secondary students aged 12 to 17 years from all Australian States and the Northern Territory. A stratified two-stage sample design was used. First, a random sample of schools was selected and second, for each school a sample of 80 students was randomly selected from predetermined year levels. A total of 351 schools participated in the survey. Students completed an anonymous, self-administered questionnaire on their smoking and drinking behaviours. RESULTS: The prevalence of current smoking (defined as having smoked at least one cigarette in the week preceding the survey) was found to increase with age to reach a peak of 25% among 16-year-old boys and 29% among girls aged 15 years. From the age of 13, smoking was more prevalent among girls than boys. Among current smokers, boys were heavier smokers than were girls. Unlike smoking, drinking was slightly more prevalent among boys than girls; boys were heavier drinkers. The proportion consuming at least one alcoholic drink in the week before the survey rose with age to a peak of 51% of boys and 46% of girls aged 17 years. Comparisons with data obtained from a similar survey conducted in 1987 showed that there had been a decrease in the proportion of 12 to 15 year olds smoking. The prevalence of drinking among both 12 to 15 year olds and 16 to 17 year olds was significantly lower in 1990 than 1987. CONCLUSIONS: While the continuing downward trends in smoking and drinking among younger students is encouraging, the results show that there are still large numbers of students smoking and drinking. PMID- 8426544 TI - Effects of walking and other exercise programs upon blood pressure in normal subjects. AB - OBJECTIVE: Rational use of non-pharmacological therapy for elevated blood pressure requires some knowledge of dose-effect relationships to optimise regimens. We investigated the effects on blood pressure of: one hour of walking at 50% of predetermined maximal work capacity (Wmax); 15 minutes of cycling at 80%-90% of Wmax (high-intensity cycling; HIC)--each performed five days per week; and three 30-minute cycling sessions per week at 65%-70% of Wmax (moderate intensity cycling; MIC) which we have previously found lowers blood pressure in both normotensive and hypertensive people. DESIGN: The three exercise interventions and a period of normal sedentary activity were performed for four weeks each, by 14 normotensive volunteers (seven male, seven female) in a randomised 4 x 4 Latin-square design. RESULTS: MIC produced the greatest blood pressure reduction relative to the period of normal sedentary activity--mean 5/3 mmHg; standard error of the difference (SE-diff) 2/1 mmHg; P < 0.05 in the supine position, and 4/5 mmHg; SE-diff 2/2 mmHg; P < 0.05 standing. Walking induced smaller blood pressure reductions--3/2 mmHg; SE-diff, 2/1 mmHg (P < 0.05 for systolic pressure), and 2/1 mmHg; SE-diff, 2/2 mmHg for the supine and standing positions, respectively. The HIC did not change blood pressure. Heart rate reduction with training was proportional to exercise intensity. Cardiac output, body weight, 24-hour urinary sodium excretion, cholesterol and triglyceride levels did not alter with any of the interventions. CONCLUSIONS: Effects of exercise on blood pressure vary according to the intensity and duration of training bouts. Moderate exercise levels may be optimal, but walking is also effective. PMID- 8426545 TI - Organochlorine pesticides in Western Australian nursing mothers. AB - OBJECTIVES: To measure the concentrations of organochlorine (OC) pesticides in nursing mothers in Western Australia and to evaluate the safety of these concentrations for breast-fed infants; to evaluate the need for future monitoring of OC pesticide concentrations and to determine whether breast milk is an accurate substitute for adipose tissue in future monitoring programs. DESIGN: Two cohorts of nursing mothers were recruited during the period October 1990 to March 1991. Levels of OC pesticides were measured in 128 samples of breast milk and 31 samples each of adipose tissue, maternal blood and cord blood. Laboratory analysis was by gas chromatography with electron capture. Health Department studies since 1974 were collated and levels of OC pesticides compared over time. RESULTS: DDT (dichlorodiphenyltrichloroethane), HCB (hexachlorobenzene) and dieldrin were found in all samples of breast milk and adipose tissue. A significant fall in the levels of OCs was noted since the 1974 survey, with the current concentrations of the cyclodienes (heptachlor, chlordane and dieldrin) being close to the limit of detection of the assay. The acceptable daily intake for dieldrin was exceeded in 90% of infants and for heptachlor in 2% of infants. The daily intakes of the other OCs were below the respective acceptable levels. HCB was detected at a median level of 0.1 mg/kg in both breast milk and adipose tissue despite being deregistered in 1972. As there is no current acceptable daily intake for HCB, the safety of this level could not be assessed by this method. A low correlation was found between levels of the cyclodiene pesticides in breast milk and adipose tissue, but levels of DDT and HCB were closely correlated. CONCLUSIONS: Restrictions on the use of the OC insecticides (DDT, aldrin, dieldrin, heptachlor and chlordane) have resulted in reduced concentrations of these chemicals in breast milk and adipose tissue as compared with previous studies. The dieldrin intake of breast-fed infants consistently exceeded the acceptable daily intake; the heptachlor intake exceeded the acceptable daily intake in some infants, but the low concentrations of heptachlor in breast milk made accurate measurement difficult. The concentrations of DDT and chlordane in breast milk did not pose a hazard to breast-fed infants. The significance of the levels of the fungicide HCB in breast milk cannot be determined. The widespread distribution of HCB, its marked persistence in the environment and its potential for human toxicity indicate a need for continued monitoring, for which breast milk is a suitable medium. The poor correlation between concentrations of dieldrin, heptachlor and chlordane in breast milk and adipose tissue may reflect the technical difficulty of measuring chemical concentrations close to the limit of detection. As aldrin is a registered pesticide and the dieldrin intake of breast-fed infants is consistently above the acceptable daily intake, monitoring of dieldrin should continue. Further population monitoring of DDT, heptachlor and chlordane is unlikely to be of value. PMID- 8426546 TI - Kidney transplantation from living related donors: a 19-year experience. AB - OBJECTIVE: To determine the outcome of patients with end-stage chronic renal failure treated by live donor renal transplantation at the Royal Melbourne Hospital and Royal Children's Hospital between 1973 and 1991, during which time two distinct immunosuppressive regimens were used. DESIGN: Data about live donor renal transplant recipients were retrieved from the Australian and New Zealand Dialysis and Transplantation Association Registry, to which we have submitted data on all transplant recipients at six monthly intervals since the commencement of our dialysis and transplant programs. PATIENTS: Seventy-two patients with chronic renal failure who received live donor renal transplants during the 19 years from February 1973 to February 1992 were included. MAIN OUTCOME MEASURES: Patient survival, transplant survival, transplant function, change in prednisolone requirements, and duration of hospital stay. RESULTS: The first 32 patients were treated with immunosuppressive regimens based on combinations of prednisolone and azathioprine ("dual therapy"), while the next 40 patients were treated with combinations of cyclosporin, prednisolone and azathioprine ("triple therapy"). Survival of patients in each group five years after transplantation was 97%. Actuarial graft survival at 5, 10 and 15 years in the dual therapy group was 58%, 52% and 47%, compared with a 5-year actuarial graft survival in the triple therapy group of 96%. There was no statistically significant difference in renal transplant function between the two groups within the first 6 years after transplantation. Twelve of 26 patients (46%) treated initially with triple therapy were able to stop treatment with prednisolone within 12 months of transplantation. Median hospital stay was 12 (range, 6-35) days during the period 1973-1985 and 8 (range, 5-20) days for the 1985-1992 period. CONCLUSION: Live donor renal transplantation has provided a highly satisfactory means of treating patients with end-stage chronic renal failure in the short and long term. Our recent experience indicates that excellent patient and graft survival and adequate renal function can be achieved by treating live donor renal transplant recipients with a triple immunosuppressive regimen of low dose cyclosporin, prednisolone and azathioprine. PMID- 8426547 TI - Microbiology laboratory clues to the diagnosis of HIV infection. AB - Microbiological investigations can provide an early clue to the possibility of unsuspected underlying HIV infection. These case studies were randomly selected from the author's clinical practice and do not represent unusual or uncommon occurrences; they highlight the constant need to ask Could it be HIV? when interpreting laboratory results. PMID- 8426548 TI - Other laboratory clues to the diagnosis of HIV infection. A key summary. AB - The variety of problems induced by HIV means that any one of a wide range of investigations may uncover the first clue to unsuspected infection. Some of the more common abnormalities reported on by pathologists and radiologists which should raise the thought of underlying HIV infection are listed in this summary. PMID- 8426549 TI - HIV infection in paediatric practice. AB - Worldwide, most new HIV infections in children are transmitted from the mother before, during or after birth. This mode of transmission is likely to become more common in Australian paediatric practice, sometimes in circumstances where the mother's HIV status is unknown and unsuspected. PMID- 8426550 TI - HIV infection in obstetric and gynaecological practice. AB - The initial impact of HIV infection on the practising obstetrician and gynaecologist was specifically related to the treatment of HIV positive women who were pregnant. The current North American experience suggests that HIV will be an important consideration in gynaecology from now on. PMID- 8426551 TI - HIV infection in surgical practice. AB - When HIV is mentioned in a surgical forum a debate will often follow about "universal testing" and "barrier precautions". Unfortunately, there is little discussion of the many ways that HIV-infected patients may present primarily to the surgeon. Although lifestyle risk factors are important clues, unusual clinical features of a relatively common surgical disease should raise the suspicion of HIV infection. PMID- 8426552 TI - HIV infection in drug and alcohol practice. AB - The injecting of illicit drugs frequently involves the sharing of injection equipment and a risk of HIV transmission. Injecting drug use at any time since 1980 should be considered a high-risk activity and users should be advised to have HIV testing. Recognition of this important lifestyle clue is more likely to occur if the doctor has evolved a practical approach to the possibility of illicit drug use. PMID- 8426553 TI - A practitioner's guide to HIV testing. AB - So far Could it be HIV? has examined many of the clues obtained from the clinical history and physical examination that might make an alert health care worker consider the diagnosis of HIV infection. Here we examine strategies for seeking permission for HIV testing and give advice on interpreting the results. PMID- 8426554 TI - Future directions in transplantation. AB - Transplantation has become an established treatment for many diseases where organ replacement is the best or perhaps the only option, but it has not reached its full potential. The use of fetal and infant donors raises ethical problems, as does the use of other species for xenografts. Legal problems include the drafting of appropriate legislation to maximise the donor pool. Economic limitations are imposed by the competition for funding with other areas of medicine. Transplantation of cells, tissues, and organs is rapidly being expanded. Exciting progress in experimental models of new methods of immunosuppression may lead to immunological tolerance of grafts so that chronic non-specific immunosuppression becomes a thing of the past. Xenografts will probably have to become a major source of organs as transplantation is extended to more patients and a broader range of diseases. Genetic engineering may alleviate the problem of supply in some instances; free cell transplants of genetically modified cells are being studied. PMID- 8426555 TI - Right or wrong? A question of answers. PMID- 8426556 TI - Compressed air diving and respiratory disease. A discussion document of the Thoracic Society of Australia and New Zealand. AB - OBJECTIVE: To review the pathophysiology and respiratory complications of compressed air diving, and to formulate guidelines for assessing respiratory fitness to dive so that diving candidates can be advised of the risks associated with respiratory disease, in particular asthma. DATA SOURCES: Specialist medical journals in the areas of respiratory medicine, physiology and diving medicine. Morbidity and mortality statistics were obtained from international diving bodies, diving medicine scientific meetings, and papers. SYNTHESIS: The major complications of underwater diving in subjects with compromised respiratory function are drowning, pulmonary barotrauma and arterial gas embolism. Diving candidates with a history of asthma, pneumothorax, obstructive or restrictive lung disease, lung cysts or thoracic trauma should be advised not to dive in view of these risk factors. CONCLUSIONS: Several respiratory diseases carry an increased risk of morbidity and mortality from compressed air diving. An accurate history and measurement of lung function are an essential part of assessing fitness to dive, both to advise potential divers appropriately and to reduce risks associated with this increasingly popular recreational activity. PMID- 8426557 TI - Planning bedside teaching--1. Overview. PMID- 8426558 TI - What do medical students know about computers? AB - OBJECTIVE: To evaluate the computer knowledge and skills of medical students in the first, fourth and final years of the Monash University medical course. DESIGN: Questionnaire. SETTING: Undergraduate medical education. PARTICIPANTS: All first, fourth and final year medical students in the Monash University medical course in 1991. RESULTS: The students in first year were found to have greater computer skills than those in sixth year. There were also significant differences between the sexes, particularly in sixth year, with male students having significantly greater computer skills than female students. CONCLUSIONS: The perceived importance of computing in medicine was high among all students and there was enthusiasm for the development and inclusion of a course on medical computing in the undergraduate curriculum. PMID- 8426559 TI - Two unusual presentations of Neisseria meningitidis infection. PMID- 8426560 TI - The economic impact of chronic fatigue syndrome. PMID- 8426561 TI - The follow-up of patients after resection for large bowel cancer, May 1992. PMID- 8426562 TI - Rural mental health. PMID- 8426563 TI - Active management of the dying patient. PMID- 8426564 TI - HIV, discrimination and health care students. PMID- 8426565 TI - Cervical self-manipulation and stroke. PMID- 8426566 TI - Tetanus in Victoria. PMID- 8426567 TI - Views of SA doctors regarding active medical help in dying. PMID- 8426568 TI - Lead poisoning due to traditional herbal preparations. PMID- 8426569 TI - Women in medicine. PMID- 8426570 TI - Oxaprozin for arthritis. PMID- 8426571 TI - Adverse prognostic features in 251 children treated for acute myeloid leukemia. AB - Potential predictors of event-free survival (EFS) were assessed in 251 consecutively diagnosed children treated for acute myeloid leukemia (AML) on three successive clinical trials. The lack of significant differences in 4-year EFS for these studies (20% +/- 4%, 29% +/- 4%, and 20% +/- 7%) permitted combined analysis of presenting features. Splenomegaly (P = .002), coagulation abnormalities (P = .001), leukocyte count > or = 10 x 10(9)/L (P = .002), and age > 14 years (P = .01) were statistically significant predictors of a poorer EFS by univariate analysis and retained significance in multivariate analysis. Age < 2 years and monocytic leukemias (often cited as adverse factors in AML) showed no prognostic influence in this study. The estimated relative risk of failure for a child with a single adverse feature at diagnosis was at least 1.4 times greater than that for a patient with no adverse features. For children with two or more adverse features, the relative risk increased by more than threefold. These clinical variables, alone or in combination, may identify important subgroups of patients with AML at high risk for failure and for whom improved or alternative therapies are especially important. PMID- 8426573 TI - Nephrotoxicity following carboplatin use in children: is routine monitoring of renal function necessary? AB - Carboplatin is an effective chemotherapeutic agent against many solid tumours. Although it is thought to be less nephrotoxic than its precursor cisplatin, current paediatric treatment protocols require regular monitoring of renal glomerular function during and after carboplatin use. Because accurate assessment of renal function in children requires measurement of isotope-clearance glomerular filtration rates (GFRs), routine monitoring is costly and time consuming. We studied 26 paediatric patients who were receiving chemotherapy that included 500-600 mg/m2 of carboplatin per course. 51Cr-EDTA GFRs were measured initially and after one to seven courses (median four). There was no measurable difference between renal function before and after carboplatin (P = 0.8). Our study shows that routine monitoring of renal glomerular function is unnecessary. Carboplatin use in patients who have had unilateral nephrectomy as part of their treatment is also discussed. Paediatric oncology literature concerning nephrotoxicity and carboplatin is reviewed. PMID- 8426572 TI - Longitudinal study of bone age in acute lymphoblastic leukaemia. AB - Bone age was assessed in children with acute lymphoblastic leukaemia (ALL) using the Tanner-Whitehouse II method. X-rays of the left hand (wrist) were made at diagnosis and annually until 5 years after diagnosis. A total of 164 X-rays from 40 patients was available. During treatment bone age development as well as height growth was retarded in relation to calendar age. A catch-up of bone age and height was observed in the 2 years after cessation of treatment. No significant differences could be found between patients with and without cranial irradiation as central nervous system treatment. Therefore cranial irradiation alone is not responsible for the growth retardation of children with ALL; the causative role of cytostatic drug therapy and/or corticosteroids has to be investigated. PMID- 8426574 TI - Allogeneic bone marrow transplantation for myelodysplastic syndromes of childhood: report of three children with refractory anemia with excess of blasts in transformation and review of the literature. AB - Myelodisplastic syndromes (MDS) in childhood deserve a negative prognosis even though disease-free survival has been obtained in 20% of cases by using aggressive chemotherapy. We describe three children with refractory anemia with excess of blasts in transformation (RAEB-T) who underwent bone marrow transplantation (BMT). We also reviewed 21 additional cases (median age was 8 years) with primary MDS recently reported in the literature with the aim of clarifying the role of BMT in treating these patients. Twelve of the 24 children were long-term survivors and free from disease at a median time of 1,320 days (range 302-2,340). There were five relapses, two graft failures, two early deaths (one VOD, one severe GVHD), and three late deaths (two respiratory diseases, one severe GVHD). We didn't find any correlation between karyotype and outcome. In conclusion, so far BMT seems to be the most valid treatment of childhood primary MDS. However, since the major causes of failure were regimen-related toxicity or recurrence of the disease after BMT, it must be pointed out that, when a compatible donor even unrelated is available, BMT for childhood MDS should be given as soon as possible or at any rate prior to blastic crisis. PMID- 8426575 TI - Evaluation of bone marrow metastasis of neuroblastoma and changes after chemotherapy by MRI. AB - To evaluate the usefulness of MRI for diagnosing bone marrow metastasis of neuroblastoma, we compared MRI findings with histological findings. MRI was performed 26 times in 20 patients with neuroblastoma to detect metastasis to the bone marrow of the femur and tibia. Abnormal areas observed by MRI were histologically examined. The lesion visualized by MRI as a low-intensity area on T1-weighted images and as a high-intensity area on T2-weighted images was histologically confirmed to be neuroblastoma in 81% (17/21). The percentage varied according to the treatment state: 89% (8/9) by MRI imaging performed before the initiation of chemotherapy, 67% (6/9) within 3 weeks after cessation of chemotherapy (during chemotherapy), and 100% (3/3) in recurrent cases 1 year or more after chemotherapy. During the follow-up period after chemotherapy, tissue with signal intensities similar to that of bone marrow was observed in a speckled pattern in the intramedullary space on T1- and T2-weighted images. This tissue was histologically demonstrated to be normal bone marrow and was considered to be bone marrow remaining after chemotherapy. In this small series, histological findings supported the results of MRI, confirming the usefulness of MRI for diagnosing bone marrow metastasis of neuroblastoma. However, bone marrow metastasis after chemotherapy was difficult to evaluate by comparing signal intensities alone. PMID- 8426576 TI - The sequellae of chemo-radiation therapy for head and neck cancer in children: managing impaired growth, development, and other side effects. PMID- 8426577 TI - Massive hematuria due to extramedullary plasmacytoma invading the bladder. AB - Bladder involvement by extramedullary plasmacytoma is a rare condition. A 46-year old woman with known IgA-lambda multiple myeloma who developed this condition is described. The patient suffered from massive hematuria, which led to hypovolemic shock. Prompt supportive treatment by blood transfusions, fulguration of the bleeding mucosa and continuous bladder irrigation stopped the bleeding. Subsequent bladder irradiation lead only to partial response of the tumor. PMID- 8426578 TI - Clostridium cadaveris bacteremia in the immunocompromised host. AB - Clostridium cadaveris, usually considered a non-pathogen, was isolated from blood cultures of two febrile patients with cancer. The bacteremias appeared to have originated from the abdomen. This organism has not been previously reported as the etiological agent in this setting. PMID- 8426579 TI - Congenital juvenile chronic myelogenous leukemia: therapeutic trial with interferon alpha-2. AB - A newborn with congenital juvenile chronic myelogenous leukemia (JCML) is described. The diagnosis was suggested by the characteristic clinical and hematologic presentation, and was confirmed by the results of in-vitro cultures of the hematopoietic progenitors, which showed excessive proliferation of monocytic colonies, with and without the addition of exogenous granulocyte macrophage colony stimulating factor (GM-CSF). Based on published in-vitro response of JCML cells to alpha interferon-2 (alpha IFN 2), we treated this child for 17 weeks with subcutaneous alpha interferon, 1,000,000 units per day. In contrast to previous in vitro results, treatment of this patient affected neither the clinical course of the disease, nor the in vitro growth of the peripheral blood-derived monocytic colonies. PMID- 8426580 TI - Increasing values of serum acid phosphatase in a child with Mycoplasma pneumoniae associated hemophagocytic histiocytic syndrome. AB - We describe a 3 1/2-year-old boy with disseminated histiocytic disease probably induced by Mycoplasma pneumoniae. In this patient, acid phosphatase was elevated in serum and was also detected histochemically in the infiltrating histiocytes. The serum acid phosphatase levels increased as his histiocytosis progressed, apparently mirroring the activity of the disease. This observation suggests that serum acid phosphatase levels should be evaluated further to determine whether they will be a useful indicator of disease in children with different histiocytosis syndromes. PMID- 8426581 TI - Treatment of childhood acute myeloid leukaemia using the BFM-83 protocol. AB - Thirty children presenting with acute nonlymphoblastic leukaemia from June 1984 to December 1989 were treated at one UK centre using a West German protocol, AML BFM-83. This consisted of Induction, an intensive outpatient-based Consolidation regimen with seven different drugs and cranial irradiation, and Continuation therapy with thioguanine and cytosine arabinoside for 2 years with additional Adriamycin in the first year. Twenty-five children achieved complete remission (83%). There were two early deaths from haemorrhage and infection and three from Induction failure. After a median follow-up time of 60 months, nine relapses have occurred, all in the bone marrow. Life table analysis revealed a probability for survival at 5 years of 47%, event-free survival 43%, and event-free interval 50%. Median bed occupancy for chemotherapy and toxicity was 41 days, with median hospital stays of 29 days for Induction, 11 days for Consolidation and less than 1 day for Continuation. This data suggests that long-term remissions can be achieved in just under half of children with acute nonlymphoblastic leukaemia while creating only modest demands on inpatient resources. PMID- 8426583 TI - A preview of the 1993 legislative session. PMID- 8426582 TI - Re: "Chemotherapy for infantile fibrosarcoma". PMID- 8426584 TI - Seventy-five years of medicine in Minnesota. PMID- 8426585 TI - The evolution of medicine as a profession. A 75-year perspective. AB - The profession of medicine has changed dramatically in 75 years. Despite the commitment of individual practitioners to the highest ideals of professionalism, the profession itself has lost privilege, power, and public reputation. It has been toppled from the high moral ground of professionalism. This has happened not so much because individual clinicians have abandoned that ground, but largely because others have occupied it--primarily complex organizations that have developed public mandates to regulate and oversee health care. The issue is not one of unfeeling physicians--it is one of a health care system that has evolved so as to limit medicine's autonomy. This changing system places new constraints and pressures on the physician-patient relationship. The question we are left with is whether medicine can regain its professionalism. How do we reform a system that, by its complexity, has become amoral? The only way is for physicians to re-assume a stout position of advocacy--advocacy for individual patients in a complex and frightening system of care, advocacy for patients as a class of people in a political system that seeks to restrict care, advocacy for patients in a world of environmental and epidemic threats. Such advocacy requires an equally strong moral commitment to the principles of service. Acting in the patient's best interest is not enough. It requires that the profession avoid the appearance of blatant self-interest at every turn. It requires a revised commitment to political activism in the interest of service to patients as a community. The costs are extremely high--but the alternative, physicians-as technicians and medicine as a slave to corporate and government interests, is hardly acceptable. PMID- 8426586 TI - Future promises revolutionary change for medicine. PMID- 8426587 TI - Health care fraud. The net widens. PMID- 8426588 TI - Health care reform: a voice for Minnesota physicians. PMID- 8426589 TI - The physician's virtues and legitimate self-interest in the patient-physician contract. AB - I will be the first to admit that we are now well into uncharted territory of the patient-physician contract. I also detect missing stretches of my dermal layer and you may spy some that I have yet to notice. In any case, I put to your serious consideration the proposal that part of the patient-physician contract must include respect for the legitimate interests of the physician by patients and third parties. The virtues of self-effacement and self-sacrifice and the concept of legitimate self-interest help us to understand in concrete, clinically applicable terms what such respect means in practice. That respect will, I think, be expressed with some variability, because there is no simple algorithm for negotiating conflicts between legitimate self-interest and the virtues of self effacement and self-sacrifice. One important consequence of this moral variability is that the patient-physician contract and the virtues that sustain it will not yield to a single, finally authoritative account of how such conflicts should be negotiated. Instead, as we turn more attention to these matters, we will, I believe, discover that there is a range or continuum of ways in which the management of such ethical conflict can reliably be understood in the patient-physician contract. Rather than a single account of the ethical dimensions of the patient-physician contract, we should expect to develop a range of reliable accounts. A kind of rich and engaging moral pluralism should thus govern our understanding of the ethical dimensions of the patient-physician contract.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426590 TI - Consumerism rampant: a critique of the view of medicine as a commercial enterprise. PMID- 8426591 TI - The necessity and the limitations of the contract model. PMID- 8426592 TI - The physician's experience: cases and doubts. PMID- 8426594 TI - The organ-tissue donation process. PMID- 8426593 TI - Whose contract and what is it? The doctor-patient agreement. PMID- 8426595 TI - Infants and others who cannot consent to donation. PMID- 8426596 TI - Debatable donors: when can we count their consent? AB - Donor consent is the standard requirement for the acceptance of an organ or tissue donation from a living donor. Usually consent to donation is respected as the choice and act of an autonomous agent. There are times, however, when the consent should be paternalistically set aside. The problem for those who must make these decisions is determining when the donor's autonomy should be respected and when paternalistic interference is appropriate. This paper draws on the writings of Kant to develop an autonomy-preserving criterion for determining where to draw the line. It then goes on to display how that criterion could be applied to the case of an adolescent donor who agrees to donate a kidney to his brother. PMID- 8426597 TI - The physician's experience: cases and doubts. PMID- 8426598 TI - Those who don't give. PMID- 8426599 TI - The contract model of the doctor-patient relationship. A critique and an alternative ethics of responsibility. PMID- 8426600 TI - Legal aspects of the sale of organs. PMID- 8426601 TI - Markets and morals: the case for organ sales. PMID- 8426602 TI - A hospital experience. AB - This article is an account of the author's experience and observations on having been incorrectly diagnosed as having a myocardial infarction and having been admitted to a coronary intensive care unit. The article touches on the rigidities of medical thinking, how one can come to believe what is not true, how to be an "exceptional patient," how medical care feels from the other side, and other relevant observations. PMID- 8426603 TI - Equality, justice, and liberty. America's unfinished agenda. PMID- 8426604 TI - Science, scientists, and responsibility. PMID- 8426605 TI - Developmental gene expression in Eimeria bovis. AB - By differential screening of stage-specific cDNA libraries of Eimeria bovis, we have identified and isolated a large set of genes that are regulated during development of the sporozoites and merozoites. Duplicate lifts of cDNA libraries constructed from partially sporulated oocysts and merozoites were probed with radioactively labeled first-strand cDNA prepared from partially sporulated oocyst and merozoite mRNA. Out of 60,000 plaques screened in each case, over 250 plaques from the partially sporulated oocyst library preferentially hybridized with the oocyst cDNA probe and 67 plaques from the merozoite library preferentially hybridized with the merozoite cDNA probe. Three of the oocyst phage and 7 of the merozoite phage were selected for further characterization. Northern analysis revealed a common pattern of mRNA expression for the oocyst cDNA clones. Consistent with the results of the differential screen, no hybridization to merozoite RNA was detected with any of these 3 oocyst cDNA clones. The expression of the merozoite cDNA clones was more complex, with 3 different classes of merozoite genes being identified based on their pattern of developmental regulation. Although each of the merozoite clones was expressed to some extent during sporulation, in all cases, expression was higher in merozoites than in partially sporulated oocysts, consistent with the restriction of expression defined by the differential screen. Sequence analysis revealed that 2 of the merozoite cDNA clones encode elongation factor 1 alpha and the ubiquitin/ribosomal protein fusion, and 1 of the sporozoite cDNAs displays a significant identity to insulin-degrading enzyme. The developmental expression of E. bovis genes involved in protein synthesis and degradation provides additional evidence for the importance of regulation of protein metabolism during parasite development. PMID- 8426606 TI - Identification of two distinct cysteine proteinase genes of Leishmania pifanoi axenic amastigotes using the polymerase chain reaction. AB - A developmentally regulated cysteine proteinase associated with an unique lysosomal organelle, the megasome, has been described for the intracellular amastigotes of the Leishmania mexicana complex; this proteinase appears to be important in the survival of the parasite. Degenerate primers encoding the active sites residues have been used to amplify cysteine proteinase cDNA sequences from axenically cultured amastigotes of Leishmania pifanoi, a member of the L. mexicana complex. Based on sequence data, two distinct genes (Lpcys1 and Lpcys2) were identified. Although both genes are preferentially transcribed in the amastigote stage, each is distinct in genomic arrangement and chromosome location, with Lpcys2 showing evidence for the presence of 8-20 tandemly arrayed copies and mRNA levels 10-fold higher than Lpcys1. Related forms of the Lpcys1 and Lpcys2 genes exist in other species of the genus Leishmania, including Leishmania braziliensis, Leishmania major and Leishmania donovani. The protein sequence of an abundant immunoaffinity purified amastigote cysteine proteinase (A 2) is identical to that predicted for the product of Lpcys2; immunofluorescence studies show an intracellular pattern/distribution for the A-2 proteinase consistent with a putative megasomal association. The DNA sequence of a genomic copy of Lpcys2 predicts a C-terminal extension for the proteinase; comparative sequence analyses of the C-terminal extensions found for Trypanosoma cruzi and Trypanosoma brucei reveal the selective conservation of cysteine, as well as proline and glycine residues, suggesting that conservation of folding and secondary structure may be required for biological function. PMID- 8426607 TI - Stable integration and expression of the Plasmodium falciparum circumsporozoite protein coding sequence in mycobacteria. AB - The DNA coding for the circumsporozoite protein of Plasmodium falciparum (CSP; aa 1-412) has been placed under the control of the mycobacterial promoter derived from the gene encoding the 64-kDa antigen of Mycobacterium bovis-BCG. This expression cassette was cloned into pJRD184, an Escherichia coli multicloning site vector, together with the kanamycin resistance gene from Tn903 and the attachment site and integrase gene from the temperate mycobacteriophage FRAT1. One of the resulting plasmids, pNIV2173, introduced by electroporation into both Mycobacterium smegmatis and M. bovis-BCG, integrated at a specific site in the genome of each recipient. Recombinants expressed immunoreactive polypeptides, ranging in size from 62 to 43 kDa, at a level of about 1% of total soluble proteins. Part of this material was present in the culture medium indicating that mycobacterial recombinants were able to secrete the CSP. The M. smegmatis and M. bovis-BCG recombinants, transformed with pNIV2173 and grown in absence of antibiotic, were followed for more than 400 and 50 generations respectively. Over this time span, neither DNA rearrangement nor loss of expression was observed. Inoculation of the recombinant BCG to mice did not induce humoral response to CSP nor proliferative response to CSP Th2R CD4+ T lymphocyte epitope. PMID- 8426608 TI - Amplification of pfmdr 1 associated with mefloquine and halofantrine resistance in Plasmodium falciparum from Thailand. AB - Drug resistance in Plasmodium falciparum is an expanding problem in most endemic areas. Recent studies have suggested the potential involvement of genes in the MDR gene family in resistance to quinoline-containing compounds in P. falciparum. In this study a molecular analysis of pfmdr 1 in recent isolates from Thailand was done (1) to further examine the role of pfmdr 1 in drug-resistant isolates and (2) to examine the reported association of pfmdr 1 intragenic alleles and chloroquine resistance. Most of the isolates (10 of 11) were resistant to all compounds tested. Analysis of pfmdr 1 revealed an apparent association between increased gene copy number and increased level of expression of pfmdr 1 and decreased susceptibility to mefloquine and halofantrine. Sequence analysis of pfmdr 1 in these isolates revealed no association of intragenic alleles with chloroquine resistance. PMID- 8426610 TI - The Ca(2+)-ATPase activity of Entamoeba histolytica is exposed towards the medium and towards the lumen of intracellular vesicles. PMID- 8426609 TI - Genomic and corrected cDNA sequence of the P28 gene from Toxoplasma gondii. PMID- 8426611 TI - Sequence of a major Eimeria maxima antigen homologous to the Eimeria tenella microneme protein Etp100. PMID- 8426612 TI - A cDNA from Schistosoma mansoni eggs sharing sequence features of mammalian cystatin. PMID- 8426613 TI - Circumsporozoite protein gene of Plasmodium simium, a Plasmodium vivax-like monkey malaria parasite. PMID- 8426614 TI - Structurally distinct genes for the surface protease of Leishmania mexicana are developmentally regulated. AB - gp63 is a highly abundant glycosylphosphatidylinositol (GPI)-anchored membrane protein expressed in both the promastigote and the amastigote forms of Leishmania species. In Leishmania mexicana, gp63 exists as a heterogeneous family of proteins that are differentially processed and localized during the 2 developmental stages. In this study we determined the molecular organization of the L. mexicana gp63 gene family, demonstrating that the gp63 genes fall into 3 linked families of tandemly repeated, but structurally distinct, entities designated as C1, C2 and C3. The C1 and C2 gene clusters contain 4-5 copies each, while the C3 gene may be single copy. Whilst promastigotes contain transcripts from all 3 gene classes, the intracellular amastigote only expresses detectable transcript from the C1 gene class. Moreover, the sequence of the C1 genes predicts a unique carboxy terminus substantially different from the GPI anchor addition signal sequence found in other Leishmania spp. and which has characteristics incompatible with substitution with a GPI anchor. These findings have significance for both the diversity of gp63 and for the regulation of tightly clustered, tandem gene arrays. PMID- 8426615 TI - Crithidia fasciculata contains a transcribed leishmanial surface proteinase (gp63) gene homologue. AB - We have cloned and analysed a gene from the insect parasite, Crithidia fasciculata, with homology to the gp63 metalloendoproteinase gene of Leishmania. The Crithidia gene homologue is arranged as a multicopy family comprised of approximately 7 genes. The mature transcript is 4.0 kb. The predicted amino acid sequence has significant homology with Leishmania gp63s, contains a zinc-binding motif and a potential site for addition of a glycosylphosphatidylinositol anchor. Demonstration of a gp63 homologue in C. fasciculata, a monogenetic parasite, suggests that the molecule may play a role in parasite survival within the insect gut. PMID- 8426616 TI - Antigenic diversity of a major merozoite surface molecule in Theileria annulata. AB - The 30-kDa merozoite surface antigen of Theileria annulata (Ankara) has been characterised by using merozoites derived from differentiating macroschizont infected cloned cell lines. This molecule was found to be 1 of the major polypeptides in extracts of both merozoites and piroplasms and was strongly detected by serum from an immune cow. Analysis of merozoite extracts derived from cloned cell lines infected with parasites of different genotypes detected 2 forms of this molecule, with respective molecular masses of 30 kDa and 32 kDa. These 2 molecules were shown to be related to each other by peptide mapping. In addition to the observed size differences, the molecules were found to be antigenically divergent. Periodate treatment of western blots demonstrated that at least 1 of the variable antigenic determinants was generated by secondary modification of the polypeptide and that a large amount of the antibody reactivity was directed against similar moieties. Parasite stocks isolated from different geographical regions were distinguished by analysis of the molecules within the 30-32-kDa molecular region. The relationship between T. annulata molecules and similar antigens described for other Theileria species is discussed. PMID- 8426617 TI - The random amplification of polymorphic DNA allows the identification of strains and species of schistosome. AB - The use of arbitrarily selected primers (10-24 nucleotides) and very low stringency annealing conditions (30 degrees C followed by 40 degrees C) for the polymerase chain reaction amplification of 1.0 ng of schistosome DNA resulted in relatively complex patterns of products. Amongst the primers tested some, for example 5'-TCGTAGCCAA, produced patterns that included bands that were polymorphic between strains of Schistosoma mansoni. Other primers, for example 5' TCACGATGCA, produced apparently identical products using DNA from 5 S. mansoni strains but highly variable patterns when DNA from different schistosome species was used. The results indicate that the random amplification of polymorphic DNA (RAPD) may be an extremely useful approach to the identification of schistosome strains and species. PMID- 8426618 TI - A 6-phosphogluconate dehydrogenase gene from Trypanosoma brucei. AB - A Trypanosoma brucei gene encoding 6-phosphogluconate dehydrogenase (6-PGDH) (EC 1.1.1.44) was identified and cloned by functional complementation of Escherichia coli gnd mutants with genomic trypanosome DNA. The T. brucei gnd gene is present as a single copy. In Northern blot experiments a probe derived from the gene hybridises to 2 transcripts (2.9 kb and 3.1 kb) which are found in both bloodstream and procyclic form organisms; the larger transcript is more abundant in bloodstream form organisms. The derived amino acid sequence of the protein is 479 amino acids in length, with a molecular weight of 52,000. It is homologous to 6-PGDHs from bacterial and mammalian sources, but diverges significantly from these other enzymes. PMID- 8426619 TI - Treatment of 171 patients with pulmonary tuberculosis resistant to isoniazid and rifampin. AB - BACKGROUND AND METHODS: The frequency of infection with multidrug-resistant Mycobacterium tuberculosis is increasing. We reviewed the clinical courses of 171 patients with pulmonary disease due to M. tuberculosis resistant to rifampin and isoniazid who were referred to our hospital between 1973 and 1983. The patients' records were analyzed retrospectively. Their regimens were selected individually and preferably included three medications that they had not been given previously and to which the strain was fully susceptible. RESULTS: The 171 patients (median age, 46 years) had previously received a median of six drugs and shed bacilli that were resistant to a median of six drugs. Thus, their regimens were frequently not optimal. Of 134 patients with sufficient follow-up data, 87 (65 percent) responded to chemotherapy (as indicated by negative sputum cultures for at least three consecutive months); 47 patients (35 percent) had no response, as shown by continually positive cultures. The median stay in the hospital was more than seven months. In a multivariate analysis, an unfavorable response was significantly associated with a greater number of drugs received before the current course of therapy (odds ratio, 4.0; 95 percent confidence interval, 1.6 to 9.9; P < 0.001) and with male sex (odds ratio, 2.5; 95 percent confidence interval, 1.1 to 6.2; P < 0.03). Twelve of the patients with responses subsequently had relapses. The overall response rate was 56 percent over a mean period of 51 months. Of the 171 patients, 63 (37 percent) died, and 37 of these deaths were attributed to tuberculosis. CONCLUSIONS: For patients with pulmonary tuberculosis that is resistant to rifampin and isoniazid, even the best available treatment is often unsuccessful. Only about half of such patients eventually have negative sputum cultures despite carefully selected regimens administered for extended periods. Failure to control this resistant infection is associated with high mortality and ominous implications for the public health. PMID- 8426620 TI - Physical fitness as a predictor of mortality among healthy, middle-aged Norwegian men. AB - BACKGROUND: Despite many studies suggesting that poor physical fitness is an independent risk factor for death from cardiovascular causes, the matter has remained controversial. We studied this question in a 16-year follow-up investigation of Norwegian men that began in 1972. METHODS: Our study included 1960 healthy men 40 to 59 years of age (84 percent of those invited to participate). Conventional coronary risk factors and physical fitness were assessed at base line, with physical fitness measured as the total work performed on a bicycle ergometer during a symptom-limited exercise-tolerance test. RESULTS: After an average follow-up time of 16 years, 271 men had died, 53 percent of them from cardiovascular disease. The relative risk of death from any cause in fitness quartile 4 (highest) as compared with quartile 1 (lowest) was 0.54 (95 percent confidence interval, 0.32 to 0.89; P = 0.015) after adjustment for age, smoking status, serum lipids, blood pressure, resting heart rate, vital capacity, body mass index, level of physical activity, and glucose tolerance. Total mortality was similar among the subjects in fitness quartiles 1, 2, and 3 when the data were adjusted for these same variables. The adjusted relative risk of death from cardiovascular causes in fitness quartile 4 as compared with quartile 1 was 0.41 (95 percent confidence interval, 0.20 to 0.84; P = 0.013). The corresponding relative risks for quartiles 3 and 2 (as compared with quartile 1) were 0.45 (95 percent confidence interval, 0.22 to 0.92; P = 0.026) and 0.59 (95 percent confidence interval, 0.28 to 1.22; P = 0.15), respectively. CONCLUSIONS: Physical fitness appears to be a graded, independent, long-term predictor of mortality from cardiovascular causes in healthy, middle-aged men. A high level of fitness was also associated with lower mortality from any cause. PMID- 8426621 TI - The association of changes in physical-activity level and other lifestyle characteristics with mortality among men. AB - BACKGROUND: Recent trends toward increasing physical exercise, stopping cigarette smoking, and avoiding obesity may increase longevity. We analyzed changes in the lifestyles of Harvard College alumni and the associations of these changes with mortality. METHODS: Men who were 45 to 84 years of age in 1977 and who had reported no life-threatening disease on questionnaires completed in 1962 or 1966 and again in 1977 were classified according to changes in lifestyle characteristics between the first and second questionnaires. We analyzed changes in their level of physical activity, cigarette smoking, blood pressure, and body weight, and the relation of these factors to mortality between 1977 and 1985. RESULTS: Of the 10,269 men, 476 died during this period (which totaled 90,650 man years of observation). Beginning moderately vigorous sports activity (at an intensity of 4.5 or more metabolic equivalents) was associated with a 23 percent lower risk of death (95 percent confidence interval, 4 to 42 percent; P = 0.015) than not taking up moderately vigorous sports. Quitting cigarette smoking was associated with a 41 percent lower risk (95 percent confidence interval, 20 to 57 percent; P = 0.001) than continuing smoking, but with a 23 percent higher risk than constant nonsmoking. Men with recently diagnosed hypertension had a lower risk of death than those with long-term hypertension (relative risk, 0.75; 95 percent confidence interval, 0.55 to 1.02; P = 0.057), as did men with consistently normal blood pressure (relative risk, 0.52; 95 percent confidence interval, 0.40 to 0.68; P < 0.001). Maintenance of lean body mass was associated with a lower mortality rate than long-term, recent, or previous obesity. The associations between changes in lifestyle and mortality were independent and were largely undiminished by age. Our findings on death from coronary heart disease mirrored those on death from all causes. CONCLUSIONS: Beginning moderately vigorous sports activity, quitting cigarette smoking, maintaining normal blood pressure, and avoiding obesity were separately associated with lower rates of death from all causes and from coronary heart disease among middle-aged and older men. PMID- 8426622 TI - Images in clinical medicine. Endoscopic laser third ventriculostomy. PMID- 8426623 TI - Management of a solitary thyroid nodule. PMID- 8426624 TI - Hodgkin's disease. PMID- 8426625 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 8-1993. A 62-year-old Cape Verdean woman with blurred vision, diplopia, a suprasellar mass, and lymphocytic meningitis. PMID- 8426626 TI - The health benefits of exercise. A critical reappraisal. PMID- 8426627 TI - Directly observed treatment of tuberculosis. We can't afford not to try it. PMID- 8426628 TI - Radiotherapy plus chemotherapy for head and neck cancer. PMID- 8426629 TI - Radiotherapy plus chemotherapy for head and neck cancer. PMID- 8426630 TI - Idiopathic hyperplasia of pulmonary neuroendocrine cells. PMID- 8426631 TI - Clinical problem-solving: treating before knowing. PMID- 8426632 TI - Clinical problem-solving: treating before knowing. PMID- 8426633 TI - More on disclosure of AIDS in celebrities. PMID- 8426634 TI - Dry eyes and video display terminals. PMID- 8426635 TI - Pseudo-red-man syndrome. PMID- 8426636 TI - Control of tuberculosis--the law and the public's health. PMID- 8426637 TI - Genes, patents and sex determination. PMID- 8426638 TI - Mixed reaction greets new gene patent proposals from Brussels. PMID- 8426639 TI - UK gene therapy gets go-ahead. PMID- 8426640 TI - Clinton lifts ban on fetal research. PMID- 8426641 TI - Inquiry into misconduct excoriates Michigan State. PMID- 8426642 TI - Montagnier and Mayor create AIDS foundation. PMID- 8426643 TI - Centocor staggered by poor clinical results. PMID- 8426644 TI - WHO vote upsets Western nations. PMID- 8426645 TI - Licences sought from PCR users in Britain. PMID- 8426646 TI - Research benefits. PMID- 8426647 TI - Genes tell a new whale tale. PMID- 8426648 TI - Earliest Homo not proven. PMID- 8426649 TI - Globin fold in a bacterial toxin. PMID- 8426650 TI - HIV tropism. PMID- 8426651 TI - HIV tropism. PMID- 8426652 TI - Revised phylogeny of whales suggested by mitochondrial ribosomal DNA sequences. AB - Living cetaceans are subdivided into two highly distinct suborders, Odontoceti (the echolocating toothed whales) and Mysticeti (the filter-feeding baleen whales), which are believed to have had a long independent history. Here we report the determination of DNA sequences from two mitochondrial ribosomal gene segments (930 base pairs per species) for 16 species of cetaceans, a perissodactyl and a sloth, and construct the first phylogeny for whales and dolphins based on explicit cladistic methods. Our data (and earlier published myoglobin sequences) confirmed that cetaceans are closely related to artiodactyls and that all families and superfamilies of cetaceans are monophyletic. A surprising finding was that one group of toothed whales, the sperm whales, is more closely related to the baleen whales than to other odontocetes. The common ancestor of baleen whales and sperm whales might have lived only 10-15 million years ago. The suggested paraphyly of toothed whales has many implications for classification, phylogeny and our understanding of the evolutionary history of cetaceans. PMID- 8426653 TI - Colour is what the eye sees best. AB - It has been argued by Watson, Barlow and Robson that the visual stimulus that humans detect best specifies the spatial-temporal structure of the receptive field of the most sensitive visual neurons. To investigate 'what the eye sees best' they used stimuli that varied in luminance alone. Because the most abundant primate retinal ganglion cells, the P cells, are colour-opponent, we might expect that a coloured pattern would also be detected well. We generalized Watson et al.'s study to include variations in colour as well as luminance. We report here that our best detected coloured stimulus was seen 5-9-fold better than our best luminance spot and 3-8-fold better than Watson's best luminance stimulus. The high sensitivity to colour is consistent with the prevalence and high colour contrast-gain of retinal P cells, and may compensate for the low chromatic contrasts typically found in natural scenes. PMID- 8426654 TI - Innervation directs receptor synthesis and localization in Drosophila embryo synaptogenesis. AB - In the Drosophila embryo, motor neurons form stereotyped synapses (neuromuscular junctions) on identified muscles. We have used a mutant (prospero) that removes or delays innervation to assay the role of the presynaptic motor neuron in the development of the receptive field of the postsynaptic muscle. prospero (pros) is not expressed in the muscles or their precursors. Here we find that the muscle defines the correct synaptic zone in the absence of the motor neuron by restricting putative guidance molecules to this specialized membrane region. Furthermore, the muscle expresses functional transmitter receptors at the correct developmental time without innervation. On the other hand, the muscle does not localize receptors to the synapse without instruction from the motor neuron, nor does a second, much larger, synthesis of receptors occur in muscles deprived of innervation. In muscles receiving delayed innervation, or muscles innervated at aberrant synaptic sites, both receptor clustering and receptor synthesis are delayed or redirected, consistent with the new pattern of innervation. We conclude that the muscle autonomously defines the synaptic site, whereas the motor neuron directs the development of the muscle's receptive field by stimulating the synthesis and localization of transmitter receptors. PMID- 8426655 TI - A new photoreactivating enzyme that specifically repairs ultraviolet light induced (6-4)photoproducts. AB - Cyclobutane pyrimidine dimers (CPDs) and pyrimidine (6-4) pyrimidone photoproducts ((6-4)photoproducts) are the two major classes of cytotoxic, mutagenic and carcinogenic DNA photoproducts produced by ultraviolet light irradiation of cells. The phenomenon of photoreactivation, the reduction of the lethal and mutagenic effects of ultraviolet radiation by simultaneous or subsequent irradiation with near ultraviolet or visible light, has been identified in several organisms and in some cases the enzymes that catalyse this process have been characterized in sufficient detail. CPDs are the only known substrate for the photoreactivating enzymes so far analysed and enzymatic photoreactivation of (6-4)photoproducts has not yet been reported. We report here that an enzyme that catalyses the light-dependent repair of (6-4)photoproduct exists in Drosophila melanogaster. This is, to our knowledge, the first report of such photoreactivating activity specific for (6-4)photoproducts in any organism. PMID- 8426656 TI - An automated approach to generating expressed sequence catalogues. PMID- 8426657 TI - AIDS cholangiopathy as the first sign of HIV infection. PMID- 8426658 TI - Psychosocial issues in pediatric AIDS. PMID- 8426659 TI - Breast cancer. Early detection may mean cure. PMID- 8426660 TI - Chronic temporomandibular joint disease and head pain. Response to surgery. PMID- 8426661 TI - The night the light went off in Sestos. Roundup (glyphosphate) poisoning. PMID- 8426662 TI - The ministry of caring, Part 2. PMID- 8426663 TI - Medicare cost shifting. PMID- 8426664 TI - The road to hell. New rules to "protect" patients from being infected with HIV by their doctors. PMID- 8426665 TI - Tuberculosis in the AIDS era. PMID- 8426666 TI - [Non-dilated urinary tract, still obstructed]. PMID- 8426667 TI - [Do ACE inhibitors help prevent heart failure? Results of 3 clinical trials]. PMID- 8426668 TI - [Controversies in preventive health care. IV. Primary health care]. PMID- 8426669 TI - [Role of C-peptide determination in the diagnosis of diabetes mellitus]. PMID- 8426670 TI - [Differences between family physicians and pulmonary specialists in the treatment of COPD patients]. AB - This cross-sectional study comprised 223 patients with moderate asthma or chronic bronchitis (FEV1 > 50% of the predicted value) from 29 general practices in the catchment area of Nijmegen University. Fifty-six patients were treated by 19 chest physicians, the remaining 167 by 29 general practitioners, without specialist care. In the study population no relevant differences in sex, age, smoking behaviour or severity of the disease (symptoms, lung function, and bronchial hyperreactivity) could be observed between the two groups of patients, except for allergy. Chest physicians prescribed on average almost three times as much medication as general practitioners. We could identify only a weak relationship between the severity of the disease (symptoms and pulmonary function combined) and the prescribed pharmacotherapy: with rising degrees of severity the general practitioner prescribes more bronchodilators, the specialist more inhaled corticosteroids. No relationship could be observed between bronchial hyperreactivity and the prescribed pharmacotherapy. No response to prescribed bronchodilators was found in 16% of the patients treated by the general practitioner and in 20% of the patients treated by the specialist. On the basis of this cross-sectional study no well-defined treatment policy of patients with moderate asthma or chronic bronchitis either by the general practitioner or by the lung specialist could be found. PMID- 8426671 TI - [Favorable effects of a stay in the Dutch Asthma Center Davos on medical consumption and quality of life in COPD patients]. AB - The Dutch Asthma Centre Davos in Switzerland is a clinic where patients with chronic nonspecific lung disease (CNSLD) are given multidisciplinary treatment. In a prospective study in the clinic, data on quality of life (functional, psychological and social characteristics) and medical consumption (use of oral corticosteroids and use of health services) were collected in a group of 147 patients with CNSLD. 18 patients were lost due to non-medical reasons. Quality of life and the use of oral corticosteroids were registered on admission, at discharge and 4 weeks, 6 and 12 months after discharge. Data on use of health services were gathered over the period between one year before admission and one year after discharge from the asthma centre. The results of this study show a decrease in the use of oral corticosteroids, in the number of visits to the family physician and outpatient department and the number and duration of hospital admissions. Favourable changes occurred in psychological functioning, (including anxiety and depression) and positive changes were observed in the degree of limitation the patients experienced in their activities of daily living. No convincing changes were found in social functioning, including social support. It can be concluded, on the basis of these results, that a stay in the Dutch Asthma Centre Davos has favourable effects on medical consumption and on some aspects of quality of life. PMID- 8426672 TI - [100 years of neurosurgery in The Netherlands]. PMID- 8426673 TI - [Increase in unsafe sex behavior among homosexual men in Amsterdam]. AB - Since 1984 a decrease in the prevalence of unprotected anogenital intercourse among homosexual men was observed in a longitudinal cohort study in Amsterdam. Two years ago, there was a strong indication that this trend was reversed, as the number of cases of gonorrhoea among homo- and bisexual visitors of the municipal STD clinic in Amsterdam started to increase again. In addition, recent analysis of behavioural data reported by participants in the cohort showed an increase in the percentage of men who had unprotected insertive or receptive anogenital intercourse (p < 0.05). Between the second half of 1990 and the second half of 1991, the percentage of men reporting unprotected insertive or receptive anogenital contact with a steady partner remained constant: 24.6% versus 27.7%. However, a serious increase occurred in the percentage of men having unprotected anogenital intercourse with casual partners (an increase from 13.1% to 24.0%; p < 0.01). Findings regarding the age distribution among homo- and bisexual men diagnosed with gonorrhoea indicate that the increase in unprotected anogenital contact was strongest among men under the age of thirty. PMID- 8426674 TI - [Financing of health research. The Dutch Heart Foundation]. PMID- 8426675 TI - [Is the use of antipyretics to lower fever and prevent febrile convulsions in children advisable?]. PMID- 8426676 TI - [Chronic subdural hematoma]. PMID- 8426677 TI - ['Mother knows best'; diagnosis of prelingual deafness]. PMID- 8426678 TI - [Illnesses come and go]. PMID- 8426679 TI - [2d Congress of the European Society of Contraception, Athens 6-9 May 1992]. PMID- 8426680 TI - Medicaid and managed care for Nebraska. A draft proposal (or one man's opinion). PMID- 8426681 TI - Management of rupture of the fetal membranes. PMID- 8426682 TI - Potential health consequences of ground-water contamination by nitrates in Nebraska. AB - Ground water serves as the primary source of drinking water for nearly all of rural Nebraska. However, ground-water contamination by nitrates, largely due to the use of fertilizers, is an increasing problem. In an ecologic study, the author found that counties characterized by high fertilizer usage and significant ground-water contamination by nitrates also had a high incidence of non-Hodgkin's lymphoma. Other potential health effects of nitrates in drinking water are also discussed. PMID- 8426683 TI - [Robotics for surgery]. PMID- 8426684 TI - [Localization of central sulcus by means of cortical SEP recordings: clinical feasibility and usefulness of intraoperative electrophysiological monitoring]. AB - Clinical feasibility and usefulness of various intraoperative electrophysiological monitoring methods that are widely utilized during neurosurgical operations were critically evaluated. We divided these intraoperative monitoring methods (IOM) into two types; the monitoring methods that enable the surgeons to obtain topographical orientation during operative procedures ('topographical IOM') and those that give them knowledge of the functional state of the nervous system that is involved in the operative procedures ('functional IOM'). It was shown why 'topographical IOM' is reliable and useful, based on our experiences of cortical SEP recordings to localize the central sulcus, and why the present 'functional IOM' is less reliable to guide the surgeons precisely during the operations. We presented some technical methods to cope with these problems in 'functional IOM'. It is necessary to establish a good correlation between the intraoperative changes of the evoked potentials and the pathological changes responsible for these electrical changes as established in clinical electrocardiography. PMID- 8426685 TI - [Long-term follow up of trigeminal neuralgic patients treated by retrogasserian rhizotomy or by microvascular decompression]. AB - The authors studied the long-term follow up of trigeminal neuralgic patients treated by retrogasserian rhizotomy with anhydrous glycerol injection (GI) or by microvascular decompression (MVD) and compared the satisfaction rates between them. Since 1983, 67 patients had been treated either by GI or MVD. The authors analyzed the results of the treatment by the method of questionnaire. Fifty-two patients of these 67 answered the questions, on which the present analysis was based. GI group consisted of 27 patients and MVD group 29 patients. The average follow-up period was 30.9 months in GI group (maximum follow-up: 84 months) and 50.9 months in MVD group (maximum follow-up: 95 months), respectively. The technique of GI used was Hartel's method with cisternography of Meckel's cave. The operations of MVD were performed by J.H. and A.K.. One patient out of 29 failed to be treated by MVD and two recurred within one year. On the other hand, 2 patients out of 27 failed to be treated by GI and 11 recurred. The recurrence was seen earlier in patients who had undergone GI, while one patient recurred the neuralgia at 76 months later. The recurrence rate in patients treated by MVD is 7.0% at 95 months, while that by GI is 49.0% at 84 months based on Kaplan-Meier survival analysis. The pathogenesis of trigeminal neuralgia is speculated to be an emphatic conduction caused by segmental demyelination and artificial synapse formation at the junction of central and peripheral myelin. The factors of this demyelination may be multiple sclerosis, basilar impression, aneurysm, arteriovenous malformation, atheroscleroses and natural aging.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426686 TI - [Acute aneurysm surgery and the Glasgow Coma Scale: relationship with 6-month outcome]. AB - A series of 610 patients who had aneurysm surgery within 7 days of the hemorrhage were analyzed as to the relationship between the preoperative Glasgow Coma Scale (GCS) score and the outcome assessed by using the Glasgow Outcome Scale (GOS) at 6 months after surgery. The patient distribution in accordance with the GCS scores in descending order from 15 to 3 was as follows: 265, 109, 44, 24, 17, 20, 25, 15, 18, 12, 16, 23, and 22 cases, respectively. In general, the larger the GCS score, the better the outcome. Thus, the overall results proved to be significantly correlated with the GCS score prior to surgery (r = 0.608, P < 0.01). As for demarcation levels along the GCS axis in terms of the GOS, a significant difference in the outcome was observed at the level of GCS scores between 15 and 14 (P < 0.0001, Wilcoxon test). However, no borderlines were evident at any GCS levels other than 15/14. The problems of applying the GCS to the grading system of aneurysmal subarachnoid hemorrhage are discussed. PMID- 8426687 TI - [Experimental study of acute spinal cord injury: a histopathological study]. AB - The microscopic appearance of a rat spinal cord which was acutely compressed by aneurysmal clip for one minute, was investigated 15 minutes, 30 minutes, 1 hour, 3 hours and 6 hours after the injury. Although the resulting small hemorrhagic lesion involved primarily only the central gray matter of the injured portion 15 minutes after compression injury, hemorrhage, necrosis and edema in the central gray matter enlarged progressively until 3 hours after injury. Petechial hemorrhage, necrosis and edema were observed in the surrounding area one hour after spinal compression. Then these pathological changes extended rostrally and caudally, but their extension was more significantly remarkable in the rostral side than in the caudal side 3 hours after compression. It was observed that leukocytes begin to infiltrate into the injured capillary walls and plug up the capillary lumen 30 minutes after the injury. Three hours following the injury, leukocytes (lymphocyte, macrophage) extravasated into the surrounding spinal tissue. From these histopathological observations, we reached the following conclusions. A. Secondary injuries play an important role in grave impairment of neurological function of the spinal cord following acute trauma. B. Pathological findings (hemorrhage necrosis and edema) extend more prominently to the rostral side, because the direction of spinal blood flow may be rostral in the thoracic spinal cord. C. Severe disturbance of intraspinal capillary blood flow leading to grave spinal damage may be evoked, because leukocytes infiltrate into the capillary around the injured area and plug up the their lumen about 30 minutes after the injury. PMID- 8426688 TI - [Vertebral artery occlusion therapy for a VA-PICA giant aneurysm: hemodynamic analysis by hydraulic vascular model]. AB - Although therapeutic occlusion of the vertebral artery has been an accepted treatment for an inaccessible VA giant aneurysm, several problems have been reported such as incomplete thrombosis, growth or rupture of the aneurysms, and cerebral embolism originating from the aneurysmal cavity. Hemodynamic changes after occlusion therapy are suspected to be responsible for these phenomena. It is usually difficult to solve these problems or to predict them before operation because multiple factors are related in a complex fashion in a living body. One of the effective means is to simulate these hemodynamic conditions by a hydraulic vascular model. To evaluate hemodynamics after gradual therapeutic occlusion of the vertebral artery, a glass sphere of 2.5 cm in diameter was placed in a hydraulic model and regarded as a VA-PICA aneurysm. 40% glycerol solution at 25 degrees C, having similar viscosity and specific gravity to human whole blood at 37 degrees C, was used as a perfusate in this study. The dye was injected into the aneurysm and intensity change of the transmitted light was measured. Half life of the dye was calculated from the thus-obtained clearance curve and was regarded as an index of intra-aneurysm stagnation. The flow volumes of each arterial site have been estimated in our previous study: 60 ml/min to the territory of one posterior cerebral artery, and 80 ml/min to the cerebellum and the brain stem. Clearance curves were recorded in the following various conditions. The flow value of the PICA was set to be 5, 10, 15, 20, 25, and 30 ml/min.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426689 TI - [Akin moyamoya disease in children]. AB - Moyamoya disease is characterized by bilateral involvement of the internal carotid arteries. The etiology of this involvement is unknown. However, we previously reported two pediatric cases of moyamoya disease that progressed from unilateral to bilateral involvement. Some cases of unilateral occlusion in the carotid fork seem to have occurred at an early stage of definite moyamoya disease. In the present study, we examined five pediatric patients showing bilateral and/or unilateral occlusion of the internal carotid artery. In each case, the etiology was known. They included Apert syndrome, radiation therapy for pilocytic astrocytoma, systemic lupus erythematosis, von Recklinghausen disease and Down syndrome. The clinical manifestations, EEG, CT, MRI, PET, and angiographic findings in these patients were presented in comparison with those of definite moyamoya disease. This comparison led us to suggest that definite moyamoya disease might be included in these cases, and we emphasize the importance of precise examination in each case. PMID- 8426690 TI - [A midbrain arteriovenous malformation at quadrigeminal plate completely obliterated by embolization]. AB - An eighteen year-old boy presented sudden loss of consciousness and tetraparesis. Radiological examinations revealed a ruptured midbrain arteriovenous malformation (AVM) at the right quadrigeminal plate. The patient recovered from symptoms by conservative treatment over 6 months with residual right hemisensory disturbance and mild diplopia during bilateral horizontal gaze. One year later, he was admitted to our institute for radical treatment of the AVM. On the angiography, the AVM was fed by two branches of the right long circumferential artery and a left paramedian penetrating artery of the posterior cerebral artery (PCA), and drained into the straight sinus via a dilated quadrigeminal vein. After much discussion on several strategies, endovascular surgery was chosen as a radical treatment. In the initial session, a medial branch of the right long circumferential artery was catheterized with Tracker-18, and embolized with 1.0 ml of ethylene vinyl alcohol copolymer (EVAL) without any deficits after negative provocative tests using Amytal and Xylocaine. Then, a lateral branch of the same artery was embolized with 0.8 ml of EVAL in the same way without any deficits. In the second session, catheterization into the left paramedian artery was difficult because of its small diameter and steep branching angle, and only a 3 mm hooked tip of Tracker-18 could be canulated into the feeding pedicle. Left oculomotor palsy was induced by provocative Xylocaine test. However, angiographic cure of the AVM was expected to be made possible by the occlusion of this pedicle, and the procedure was continued due to the request of the patient and his family.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426691 TI - [A case of posterior cerebral artery giant aneurysm with xanthochromic content]. AB - We reported a case of posterior cerebral artery giant aneurysm with xanthochromic content. A 28-year-old male patient was admitted to our department suffering from lapses into unconsciousness once a month for the previous two years and a half. CT scan demonstrated a mass 2 cm in diameter in the right ambient cistern. The density of the inside of the mass was low, and that of outer border was high. The mass was not enhanced. MRI demonstrated that the intensity of the inside of the mass was low, and the outer part was iso-low in the T1 weighted image, while the inside was high and the outer part low in the T2 weighted image. Cerebralangiography showed an aneurysm of 5 mm in diameter at the bifurcation of the posterior cerebral artery (PCA) and the posterior inferior temporal artery. There was occlusion of the PCA. Accordingly, we diagnosed it as a calcified giant aneurysm with fluid-like content, and carried out surgery using the extended subtemporal approach. Inside the very stiff yellow-white wall of the aneurysm, there was only xanthochromic fluid without thrombus, and the structure of the aneurysm was cystic. The aneurysm was resected following neck clipping. After the operation, the patient's lapses into unconsciousness were able to be well controlled with anticonvulsants, and he left our department without any neurological deficit about a month afterwards. The histological findings of the specimen show the inner wall of the artery. Cases of giant aneurysms with xanthochromic content have rarely been reported. Only two are recorded, and this is the first case diagnosed using MRI.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426692 TI - [Ruptured cerebral aneurysm associated with chronic renal failure: case report and evaluation of dialysis]. AB - Two cases of ruptured cerebral aneurysm with chronic renal failure were successfully treated by selecting an appropriate dialysis during the pre- and postoperative period. Case 1; a 41-year-old male, who had been receiving hemodialysis for 4 years, complained of sudden onset of headache, and his consciousness deteriorated abruptly afterwards. A ruptured basilar-left superior cerebellar artery aneurysm was diagnosed, and an external ventricular drainage device was installed. The patient slowly recovered consciousness and was scheduled for delayed operation. During this period hemodialysis was suspended and continuous ambulatory peritoneal dialysis (CAPD) was performed instead. On the 24th day, the aneurysm was clipped, and CAPD switched to ordinary hemodialysis three days after the operation. However, consciousness deteriorated and CT scan showed diffuse cerebral swelling due to disequilibrium syndrome. The patient recovered consciousness 24 hours after hemodialysis. Frequent short-term dialysis eventually eliminated this syndrome. Case 2; a 60-year-old male, who had been receiving hemodialysis for 6 years, complained of a sudden severe headache, and a ruptured anterior communicating artery aneurysm was diagnosed. Emergency clipping of the aneurysm was performed and, except for mild vasospasm on the seventh day, the patient's recovery was uneventful postoperatively, with non anticoagulative agent hemodialysis. These two cases demonstrate that chronic renal failure of a ruptured cerebral aneurysm patient can, with good prognosis, be treated by CAPD preoperatively, and non-anticoagulative agent hemodialysis postoperatively, followed by ordinary dialysis carefully avoiding the disequilibrium syndrome. PMID- 8426693 TI - [History and advances of measurement of cerebral blood flow and metabolism: from cranial window technique to near infra-red CT (series 1)]. PMID- 8426694 TI - [Surgical removal of lateral pontine cavernous angioma: review of the surgically treated cases in the literature]. AB - A case of cavernous angioma of the pons which was surgically and successfully excised was reported. A 36 year-old man complained of progressive headache, double vision and tinnitus. Neurologic examination revealed left fifth, sixth and seventh cranial nerve palsies. He had left limb ataxia and right sided hemisensory deficit. A computed tomographic (CT) scan on admission disclosed a hematoma in the left lateral portion of the pons. Serial CT scans demonstrated progressive increase of hematoma. MRI scans revealed an area of mixed signal intensity in T1 weighted images. These findings were thought to be consistent with a cavernous angioma. Three months after the onset, surgery was performed using a lateral suboccipital approach. Histological examination disclosed cavernous angioma. After surgery, the patient's neurological deficits improved, and after 3 months, all symptoms except the mild limb ataxia had disappeared. PMID- 8426695 TI - [Multiple meningiomas of thoracic spinal cord: report of two cases]. AB - Multiple spinal meningiomas are relatively rare and account for from only 2 to 3.5% of all spinal meningiomas. Two cases of multiple meningiomas of the thoracic spinal cord were reported. Case 1. A 73-year-old woman was admitted with a 5 month history of progressive motor weakness and sensory disturbance in the legs bilaterally. Neurological findings on admission revealed paraparesis, hypesthesia and hypalgesia under the Th10 level, hyperreflexia of both legs, and urinary incontinence. Myelography and MRI revealed two intradural extramedullary lesions at the Th7 and Th10 level. Both tumors were removed completely. Histopathological examination showed psammomatous meningiomas. Case 2. A 52-year-old man was admitted with a 2 month history of progressive motor weakness and numbness of both legs. Neurological findings on admission revealed paraparesis, hypesthesia and hypalgesia under the Th10 level, hyperreflexia of both legs, and genitourinary incontinence. Lumbar myelography showed complete block at the Th9 level. MRI showed stenosis of the spinal canal at the Th8/9 level, and a deviation of the spinal cord. MRI with Gd-DTPA showed that the spinal cord was compressed by intradural extramedullary tumors. However, myelography and MRI could not detect the multiplicity of tumors. CT myelography demonstrated three separate tumors from the Th7 to the Th9. Three tumors were totally removed. Histopathologically, they were meningotheliomatous meningiomas. These cases were considered as being multiple meningiomas of the spine. Both patients showed no manifestations of von Recklinghausen's disease. The cause of the multiplicity in these cases was uncertain. Multiple spinal tumor is very difficult to diagnose because of unusual clinical symptoms.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426696 TI - [History of moyamoya disease]. PMID- 8426697 TI - A greater reward. PMID- 8426698 TI - Faulty conclusion. PMID- 8426699 TI - For-profit referral services: they're illegal, aren't they? PMID- 8426700 TI - Stress management in pediatric dentistry. PMID- 8426701 TI - Radiography for children and adolescents. PMID- 8426702 TI - Breastfeeding and oral health. A primer for the dental practitioner. PMID- 8426703 TI - Behavior management techniques in pediatric dentistry. AB - There are many techniques a dentist can use to aid in delivering care to young patients. Proper selection and use can affect the immediate visit as well as future visits. PMID- 8426704 TI - When an insurance carrier requests an X-ray. PMID- 8426705 TI - National Children's Dental Health Month: keystone of preventive dentistry. PMID- 8426706 TI - Dental allowance. PMID- 8426707 TI - Setting dentists free. PMID- 8426708 TI - Male subfertility due to sperm antibodies: a clinical overview. PMID- 8426709 TI - Workplace stress rated for AIDS caregivers. PMID- 8426710 TI - Workplace injury/illness rates for 1991 show a 10-year record drop in incidence. PMID- 8426711 TI - Descriptive terminology for jaw anomalies. AB - A descriptive terminology for jaw abnormalities was developed and has been used for many years. This terminology was based primarily on inspection of patients and evaluation of cephalometric x-ray images and defines the anatomic location of abnormalities. Its aids in communication between doctors of different specialties. PMID- 8426712 TI - Clinical observations of cleft lip and palate. AB - In surgical management of cleft lip and palate, appreciation of the special anatomy and physiology of the human premaxilla is more important than the question of whether it is a separate bone. To take advantage of the potential of the premaxilla, both primary and secondary surgical procedures must strive to establish a nearly normal medial septal system and nasolabial musculature. To complement accurate muscle surgery of the lip and soft palate, the characteristics and functions of the mucoperiosteum that covers the palate must be respected. The quality of subsequent facial growth can be monitored with the architectural craniofacial cephalometric analysis. By adopting a physiologic approach to cephalometrics, the clinician can avoid some common errors of interpretation. PMID- 8426713 TI - Fine-needle aspiration biopsy in oral and maxillofacial surgery. AB - Fine-needle aspiration biopsy is a technique that is gaining great popularity in both surgical and nonsurgical specialties. Its accuracy, safety, and usefulness have been demonstrated repeatedly; however, its usefulness seems to have been overlooked by oral and maxillofacial surgeons. This technique has many advantages in the diagnosis of mass lesions, including ease of use, cost effectiveness, convenience, and accuracy. It lends itself well to oral and maxillofacial surgery. PMID- 8426714 TI - Comparison of oral triazolam and nitrous oxide with placebo and intravenous diazepam for outpatient premedication. AB - Triazolam was evaluated as an oral sedative agent for dental outpatients in two studies in the oral surgery model. The first study demonstrated that 0.25 mg of triazolam in combination with nitrous oxide provides therapeutic effects but with a more rapid recovery than a 0.50 mg dose in combination with nitrous oxide. In the second study, triazolam produced a significant anxiolytic effect that was comparable to the effects of diazepam titrated to the usual clinical endpoint (mean dose = 19.3 mg). Less impairment in cognitive-psychomotor impairment and ambulatory function was seen after triazolam in comparison with diazepam. Triazolam appears to be a safe, effective alternative to parenteral sedation with a benzodiazepine for dental outpatients. PMID- 8426715 TI - Complications of dental surgery in persons with HIV disease. AB - Post tooth extraction infective complications have occasionally been described in HIV-infected persons. However, there is little objective data as to the frequency of this and the need for antibiotic prophylaxis. Similarly the frequency of postextraction bleeding in patients infected with HIV, who may have thrombocytopenia, is unknown. In the present study the frequency of postextraction complications has been investigated in a group of 38 persons at stages 2 to 4 of HIV infection and 26 matched subjects from patients groups commonly at risk of HIV infection but not seropositive. During 40 clinical procedures in HIV-infected patients, 100 (range 1 to 23, median 1) teeth were extracted. Three episodes of delayed postextraction healing were recorded. During 30 procedures in the non-HIV-infected persons, 68 (range 1 to 5, median, 2) teeth were extracted, and two episodes of delayed postextraction healing were recorded. These differences were not significant. Only one HIV-infected patient had an episode of severe postextraction bleeding: this was a hemophiliac who bled despite receiving factor VIII prophylaxis. The bleeding occurred 7 days after the extraction and ceased with tranexamic acid and additional factor VIII. No control subject had severe postextraction hemorrhage. It is concluded that postextraction complications are uncommon in HIV-infected patients and that routine antibiotic prophylaxis is not indicated. PMID- 8426716 TI - Class I and II HLA antigens in British patients with oral lichen planus. AB - The frequencies of human leukocyte antigens--HLA-A, HLA-B, HLA-C, HLA-DR, and HLA DQ1--were determined in a group of 40 white British patients with oral lichen planus and compared with those of healthy controls. Alterations in the frequencies of several HLA antigens were noted. In particular, an increase in HLA Bw57 and a decrease in the frequency of HLA-DQ1 were seen in the group with lichen planus. When different clinical subgroups of lichen planus were compared with the control group, significant changes were also noted in the frequencies of HLA antigens. This suggests that lichen planus may represent a heterogeneity of diseases and that HLA-Bw57 may predispose a person to lichen planus whereas HLA DQ1 may be associated with resistance to it. PMID- 8426717 TI - Fluocinonide in an adhesive base for treatment of oral lichen planus. A double blind, placebo-controlled clinical study. AB - Patients with symptomatic oral lichen planus frequently require therapy to reduce signs and symptoms. For this purpose, corticosteroids are often applied topically. In a randomized, double-blind, placebo-controlled study, the efficacy of the topical application of 0.025% fluocinonide was evaluated. Forty consecutive patients with oral lichen planus diagnosed on the basis of histopathologic and immunofluorescence findings participated in this study. All patients were followed for 3 to 17 months. No adverse effects were noted during follow-up period. In the group of 20 patients that received the drug, 4 patients (20%) showed a complete remission, and 12 patients (60%) had a good or partial response to topical treatment. In the placebo-group, these figures were 0 and 6 (30%), respectively. The majority of the placebo-group (70%) did not respond at all with regard to signs (Xt2 = 10.4; p = 0.0013) and symptoms (Xt2 = 6.97, p = 0.008). The results from this study suggest that topical application of fluocinonide in an adhesive base is a safe and effective drug to reduce signs and symptoms in oral lichen planus. PMID- 8426718 TI - Treatment of oral Kaposi's sarcoma with a sclerosing agent in AIDS patients. A preliminary study. AB - Presently oral Kaposi's sarcoma is primarily treated with systemic and intralesional chemotherapy, immunotherapy, radiation, and occasionally lasers. Each of these modalities achieves varying degrees of success. In patients with acquired immunodeficiency syndrome, the traditional treatments are frequently accompanied by local and generalized side effects that are detrimental to an already compromised immune system. Experience using sclerosing agents to treat other oral vascular lesions in healthy patients is known to produce excellent results. To evaluate the use of this treatment method in patients with acquired immunodeficiency syndrome, 15 oral lesions in 12 patients were injected with 3% sodium tetradecyl sulfate (Sotradecol). Thirteen lesions were located on hard palate; nine were nodular, three papular, and three macular. One week after treatment, some degree of ulceration occurred in all lesions with healing beginning by the middle of the second week. Six lesions required a second treatment to completely resolve. In one patient, healing was delayed, and the patient encountered a period of discomfort as a result of superficial bone sequestration. All other lesions healed uneventfully without the major side effects encountered with the other commonly used methods. Patients were followed for recurrence until they died or moved away. No recurrences were noted, with many followed for 18 months or longer. It is suggested that for lesions 2.6 cm or less in size, a sclerosing agent may be a better treatment modality than those now commonly used for oral Kaposi's sarcoma. PMID- 8426720 TI - Paraneoplastic pemphigus. A distinct autoimmune vesiculobullous disorder associated with neoplasia. AB - A vesiculobullous disease termed paraneoplastic pemphigus with distinct autoantibodies was newly described in 1990. All reported cases have occurred in patients with a history of neoplasia, including lymphoma, chronic lymphocytic leukemia, poorly differentiated sarcoma, and benign thymoma. As in pemphigus vulgaris, intraepithelial clefts with acantholysis are noted histopathologically, and intercellular binding of immunoreactants is seen with direct immunofluorescence studies of mucous membrane and skin biopsies. However, immunoreactants may also be found along the basement membrane zone in paraneoplastic pemphigus. Indirect immunofluorescence using rat bladder epithelium as substrate shows an intercellular pattern that appears to be highly specific for paraneoplastic pemphigus. We report a patient with non-Hodgkins lymphoma of 8 years duration who developed severe erosive stomatitis and lichenoid dermatitis after receiving chemotherapy for a relapse of lymphoma. Her case illustrates the typical features of the disorder described as paraneoplastic pemphigus. Neoplasia-associated pemphigus may be a more precise term for this disorder because the course of the blistering eruption does not always parallel the course of the underlying cancer. The clinical features, histopathologic findings, and immunofluorescence findings of this unique syndrome are reviewed. PMID- 8426719 TI - Central giant cell lesions of the jaws. A clinical, radiologic, and histopathologic study. AB - The histology, radiographs, and follow-up information for 142 cases of central giant cell lesions of the jaws were reviewed in an effort to determine which, if any, microscopic features could be correlated with clinical behavior. The majority of these lesions were asymptomatic and relatively innocuous. However, some displayed a more aggressive clinical course characterized by root resorption, pain or paresthesia, and cortical perforation. The over-all recurrence rate in the 142 cases was 16%. Adequate follow-up information (mean, 48 months) was only obtained for 47 patients, and 23 (46%) of these experienced one or more recurrences. Statistically significant histologic differences in distribution of giant cells and frequency of osteoid within the lesions were found in lesions that recurred as opposed to those that did not. The concept that giant cell lesions of the jaws are not totally different entities from giant cell tumors is discussed. PMID- 8426721 TI - Malignant transformation of peripheral ameloblastoma. AB - A peripheral (extraosseous) ameloblastoma was excised from the maxillary left tuberosity of an 82-year-old man. It recurred twice over a 5-year period, once as squamous cell carcinoma and again as undifferentiated carcinoma. Analysis of the literature suggests that peripheral ameloblastomas may have the same potential for malignant transformation as intraosseous ameloblastomas. PMID- 8426722 TI - Oral Crohn's disease and pyostomatitis vegetans. An unusual association. AB - Oral features of Crohn's disease include ulcers, lip fissuring, cobblestone plaques, angular cheilitis, polypoid lesions, and perioral erythema. Pyostomatitis vegetans is a rare eruption of the oral mucosa characterized by tiny yellow pustules. It is considered a marker for inflammatory bowel disease. We describe a 45-year-old woman with a 6-month history of painful sores in her mouth, diarrhea, weight loss, and cutaneous lesions. Oral examination revealed cobblestone plaques and indentation on the tongue and friable vegetating pustules on the labial commissures. Staphylococcus simulans was isolated from the pustules. Laboratory studies revealed leucocytosis, eosinophilia, and low hemoglobin and zinc levels. Histologic study of the labial lesions revealed hyperplastic epithelium with intraepithelial clefts that contain eosinophils and neutrophils. Tongue lesions showed chronic inflammation with noncaseating granulomas. Later, colonoscopy and biopsy demonstrated Crohn's disease of the anorectal region. Pyostomatitis vegetans lesions regressed after oral zinc supplementation. Prednisone treatment resulted in healing of the tongue lesions. In our patient, pyostomatitis vegetans appeared to be related to zinc deficiency that may have been caused by malabsorption. The pathogenetic interrelationship between pyostomatitis vegetans and Crohn's disease is discussed. PMID- 8426723 TI - Is phantom tooth pain a deafferentation (neuropathic) syndrome? Part II: Psychosocial considerations. AB - The symptoms of phantom tooth pain are often considered to be of psychological origin by those unfamiliar with its clinical characteristics. Part of the problem is that phantom tooth pain is often confused with atypical facial pain. Extensive literature exists for the latter that suggests a psychological cause. Many studies of atypical facial pain, however, suffer from one or more of four methodologic problems. In the present study, 115 phantom tooth pain cases are compared with a contrast group of 151 facial pain cases and 137 nonpain controls on a variety of personality characteristics. Only one trait personality factor, locus of control, statistically differentiates the three groups. The chief psychological difference of the phantom tooth pain sample compared with the control and contrast groups was higher scores on a measure of demoralization. Demoralization can be interpreted as a consequence as well as an antecedent of the chronic pain state. Evidence for each opinion is presented. Suggestions for obtaining informed consent of prospective endodontic patients are suggested. This study has not demonstrated that phantom tooth pain cases are characterized by a specific premorbid personality. PMID- 8426724 TI - Treatment and prognosis of avulsed teeth. A discussion and case report. AB - A documented case that involves avulsion of an incisor is reported. Calcium hydroxide failed to prevent progressive irreversible root resorption. The importance of extraoral time, transport media, duration and type of splinting, as well as the rationale for the use of calcium hydroxide in the treatment of avulsed teeth are discussed. This case could serve as a reminder to practitioners, that patient compliance, timing and sequencing of endodontic therapy, and regular recalls is critical in the prevention or treatment of root resorption in avulsed teeth. It also emphasizes the frustration of many clinicians in treating avulsed immature teeth. The reporting of long-term documented cases should be encouraged so that replantation techniques may be evaluated. PMID- 8426725 TI - Assessing condylar changes with digital subtraction radiography. AB - Transcranial radiographs of the temporomandibular joint with and without simulated pathology were compared with digital subtracted and histogram equalized images of the same joints. Subtracted images had specificity and sensitivity values of 0.83 and 0.76 respectively, compared with values of 0.42 and 0.54 for conventional radiographs. It was concluded that digital subtraction radiography has the potential to increase the diagnostic yield of transcranial temporomandibular radiography for bony changes to the condylar head. PMID- 8426726 TI - Clinical correlation of oral-dental findings with radiographs and with total body bone scans. AB - Bone scans are frequently used to detect osteoblastic areas in bone, including bony metastases in patients with existing tumors. Various dental conditions have been found to cause areas in the jaws to have increased uptake of radiopharmaceuticals. We studied 30 patients with an existing cancer or previous history of cancer with the use of total body bone scans, panoramic radiographs, and dental examinations, and we found no correlation between the intensity of radionuclide uptake in the jaws on the bone scans and the number of teeth in each jaw, the age of the patient, the degree of periodontal disease, or the number of dental pathoses per jaw. The frequency and intensity of positive scan results were related to the presence or absence of intrabony lesions in the jaws. Dental disease therefore does not appear to mask metastatic disease in the jaws; however, when metastasis is suspected, a dental examination with radiographs is recommended. PMID- 8426727 TI - Unusual delayed sequel to facial trauma. PMID- 8426728 TI - [Management of arteriosclerosis-related lipid metabolism disorders. Recommendations of the Hungarian Lipid Consensus Conference]. PMID- 8426729 TI - [Helicobacter detected in children of Helicobacter pylori-positive parents]. AB - The authors examined 48 children of Helicobacter pylori positive parents: 52% were serologically positive for Helicobacter pylori using the Orion Diagnostics Pyloriset Latex agglutination test. Of these children 10 had upper abdominal pain, heartburn and acid eructation and osesophagogastroscopy was performed; seven were found to be Helicobacter pylori positive histologically. These results draw attention to the transmission of Helicobacter pylori infection within families. They suggest that these children should be reviewed regularly, and the diagnostical examinations need to perform if the clinical picture is suspicious of Helicobacter pylori infection. PMID- 8426730 TI - [Multiple primary malignant tumors involving the liver]. AB - In the Department of Pathology of the Albert Szent-Gyorgyi Medical University in Szeged during the last 30 years 1770 (19.4% of the cancers) primary malignant lung tumours were observed in autopsy material, from which 86 patients (4.9%) had other malignancies as well. In 81 cases other extrapulmonary and in 5 cases other primary lung tumours were observed. The male predominance in these cases was significant. All of the patients were heavy smokers. Amongst these synchronous tumour-associations the most frequent extrapulmonary tumours arose in the urogenital tract, in the head and neck, relatively frequently also in the breast, liver, stomach, intestine and thyroid. These cases caused diagnostic dilemmas both for the clinician and even for the pathologist. Several signs help to distinguish a new primary tumour from a metastasis. Multiplicity itself does not mean poorer prognosis. Each cancer should possibly receive adequate treatment. PMID- 8426731 TI - [Hearing loss resulting from purulent meningitis in the light of adjuvant dexamethasone therapy]. AB - Results of objective audiometry of 109 infants and children after purulent meningitis are presented. Among them 17 patients got dexamethasone as a supportive therapy. There was no statistically significant difference in hearing loss between the dexamethasone-treated and the control group. (41 vs 43% sensorineural hearing loss respectively). Authors do not contraindicate the dexamethasone therapy in purulent meningitis because of its harmlessness and useful effect to the course of the disease but further investigations are needed for avoidance of hearing loss following meningitis. PMID- 8426732 TI - [Sneddon syndrome as a clinical manifestation of the antiphospholipid antibody syndrome]. AB - A 39 year old female patient has had reticular skin lesions showing the clinical picture of livedo racemosa for 7 years. Her medical history revealed cerebrovascular disease. In the background of livedo racemosa no systemic disorder could be detected, so the authors considered their case to be Sneddon's syndrome. In the serum of the patient antiphospholipoid antibodies (high anticardiolipin antibody titer, factor lupus anticoagulant) could be detected so the diagnosis mentioned in the title seemed to be correct. PMID- 8426733 TI - [Lyme borreliosis]. PMID- 8426734 TI - GM-CSF receptor: interactions and activation. PMID- 8426735 TI - The Adenovirus E1A gene blocks the differentiation of a thyroid epithelial cell line, however the neoplastic phenotype is achieved only after cooperation with other oncogenes. AB - The PC Cl 3 cell line is a well characterized epithelial thyroid cell line of Fischer rat origin. This cell line has the peculiarity of retaining in vitro the typical markers of thyroid differentiation (i.e. thyroglobulin synthesis and secretion, iodide uptake and dependence on TSH for growth). The PC Cl 3 cells have been transfected with the E1A gene of Adenovirus 5. The E1A transfected cells, PC E1A, partially lost the dependency on TSH for growth and completely lost the ability to trap iodide and synthesize thyroglobulin; however they did not acquire the typical markers of the neoplastic phenotype. A highly malignant phenotype was achieved after infection of the PC E1A cells with retroviruses carrying the v-raf, v-abl and polyoma virus middle T oncogenes. In contrast, the PC E1A cells transfected with the E1B gene of Adenovirus were not tumorigenic at all, and those infected with retroviruses carrying oncogenes of the ras family displayed a very weak tumorigenic phenotype. PMID- 8426736 TI - Expression of antisense E1A in 293 cells results in altered cell morphologies and cessation of proliferation. AB - To determine whether continued E1A expression is required to maintain immortalization, 293 cells, a cell line that has been immortalized and transformed by adenovirus E1A and E1B, respectively, were transfected by an antisense E1A expression vector. In the presence of low serum, 293 cells underwent a transient, viable, nonproliferative phase wherein cells with altered morphologies were detected. However, only those clones which did not integrate the antisense sequences and exhibited a morphology similar to the original 293 cells were able to survive. High serum concentrations resulted in fewer clones, which rapidly discarded the antisense DNA. Similar experiments with an antisense CAT vector did not give rise to cells with either altered morphologies or growth rates. Furthermore, CAT antisense DNA was integrated into the 293 cell DNA. Hela cells, which were not immortalized or transformed by E1, were unaffected by the expression of antisense E1A and integrated the antisense DNA into genomic DNA. Consistently, the addition of synthetic antisense, but not sense oligonucleotides resulted in a transient inhibition of 293 cell DNA synthesis. These data demonstrate that even after extended periods of time in culture, cells immortalized by E1A still require E1A expression to activate the cell cycle and prevent them from senescing. PMID- 8426737 TI - Tumor suppressor activity of RB and p53 genes in human breast carcinoma cells. AB - Breast cancer is the most common cancer of women in Western countries. Various genetic alterations have been implicated in its development. Two tumor suppressor genes, the retinoblastoma susceptibility gene (RB) and the gene encoding the p53 protein, are frequently found to be deleted or mutated in breast cancer cell lines and primary tumor samples. Breast carcinoma cell lines MDA-MB468 and BT549 both harbor partial RB gene deletions as well as point mutations of their p53 genes, thus providing an excellent model system for testing the roles played by these two genes in the oncogenesis of breast cancer. Single copies of wild-type RB or p53 were delivered to these cells by retrovirus-mediated gene transfer. Restoration of RB expression in cells reduced their ability to grow in soft agar and their tumorigenicity in nude mice, although no significant alteration of growth rate in culture could be detected. Introduction of wild-type p53 into these cells exerted a similar effect on the neoplastic phenotypes. This effect occurred even in the presence of their endogenous mutated p53 alleles, which argues for the phenotypical dominance of the wild-type p53 over mutated p53 during human oncogenesis. The ability of RB and p53 genes to suppress the tumorigenicity of breast carcinoma cells provides functional evidence that deletion or mutational inactivation of tumor suppressor genes represents an important step in the genesis of breast cancer. PMID- 8426738 TI - p53 and K-RAS alterations in pancreatic epithelial cell lesions. AB - We have analysed the expression of p53 at the mRNA level, and extensively at the protein level by immunostaining, Western blotting, and ELISA measurements revealing a p53 increase in 8 out of 14 cell lines established from human pancreatic carcinomas. The mRNA levels closely paralleled the protein levels in most of the cell lines. Overexpression of p53 in tumor cells correlated with mutations in the p53 gene. Immunocytochemistry was also performed with tissue cryosections showing a nuclear p53 staining in 8 out of 12 exocrine, and 2 out of 2 endocrine tumors. In addition, nonmalignant peri-tumoral tissue specimens and cells derived from pancreatic juice of acute pancreatic patients were also positively stained. These findings may suggest functions of p53 in stress situations induced by acute inflammation or tissue regeneration. Genomic mutations in the tumor suppressor gene were associated with point mutations in either codon 12, 13 or 61 in the c-K-RAS oncogene in about two-thirds of cell lines. The frequent activations of a RAS oncogene in combination with mutations of a tumor suppressor gene are likely to contribute to the malignant phenotype of pancreatic adenocarcinomas. PMID- 8426739 TI - A functionally inactive p53 Li-Fraumeni syndrome mutant. AB - Germline mutations in the tumor-suppressor p53 have been recently identified in Li-Fraumeni syndrome patients. We analysed the function of one of these mutations, an arg-to-trp substitution at amino acid 245 in the murine p53 gene. This p53LFS mutant could not, unlike wild-type p53, suppress foci formation of rat embryo-fibroblasts. Like other p53 mutants it cooperated with activated ras to transform rat embryo fibroblasts. Overexpression of p53LFS thus resulted in a phenotype similar to other mutant p53s. The p53LFS protein was also transcriptionally inactive in contrast to previous studies using a p53LFS/GAL4 fusion protein. To better understand the functional domain disrupted in p53LFS, we developed a dimerization assay and showed that p53LFS still dimerized. In addition, p53LFS retained its ability to bind SV40 large T antigen and not hsc70, both characteristics of wild-type p53. Using immunofluorescence, we localized p53LFS to the nucleus. From these results we conclude that p53LFS represents an unusual p53 mutant in that it retains many characteristics of wild-type p53, however activities critical for growth suppression are lost. PMID- 8426740 TI - Induction of nuclear accumulation of the tumor-suppressor protein p53 by DNA damaging agents. AB - Cancer therapy drugs, such as diamminedichloroplatinum (cisplatin), mitomycin C, etoposide and a number of other compounds, as well as energy-rich radiation, are known to act on cellular DNA. These agents are shown to induce nuclear accumulation of the so-called tumor-suppressor protein p53 in fibroblastoid cells, as well as in epithelioid normal and immortalized cells of murine, simian, and human origin. p53 accumulation starts a few hours after treatment and can remain detectable in surviving cells for at least 20 days. Accumulation occurs because of increased p53 protein stability and depends on ongoing translation. It is not the result of enhanced gene expression. A number of cell cycle inhibitors do not affect p53 protein accumulation, suggesting that the process may start from several points in the cell cycle. Since the increase in the nuclear p53 protein levels occurs within a few hours in most of the treated normal diploid cells, it is unlikely that the accumulated p53 protein is derived from a mutated p53 gene. The results obtained are in accordance with the view that the DNA damage-induced p53 accumulation may either inhibit cell growth, allowing DNA repair processes, or, in the case of severe damage, initiate apoptosis. PMID- 8426741 TI - Hypoxic stress induces reversible hypophosphorylation of pRB and reduction in cyclin A abundance independent of cell cycle progression. AB - The effect of hypoxic stress on the phosphorylation state of the product of retinoblastoma susceptibility gene (pRB) and cyclin A abundance was examined in CV-1P monkey kidney cells. Flow cytometric DNA histogram analysis and [3H] thymidine incorporation assays demonstrated that hypoxia inhibited cell cycle progression and cell division. Within 6-12 h of hypoxia, pRB became hypophosphorylated and cyclin A abundance fell below detection limits. Hypophosphorylation of pRB and loss of cyclin A detection occurred without progression of cells through S-phase. These effects were found to be reversible by reoxygenation of the hypoxic cultures. Cells were shown to resume DNA synthesis within 12-16 h of reoxygenation concomitant with pRB hyperphosphorylation and an increase in cyclin A detection. These data demonstrate that hypoxic stress blocks the progression of these cells through the phases of the cell cycle and suggests that the effect might arise from the down regulation of key cell cycle controlling elements. The data also raise the possibility that maintaining pRB in a hyperphosphorylated state may be crucial for S-phase progression as well as S-phase entry. PMID- 8426742 TI - Raf revertant cells resist transformation by non-nuclear oncogenes and are deficient in the induction of early response genes by TPA and serum. AB - A revertant cell line was generated from v-raf transformed NIH/3T3 fibroblasts. These cells, termed CHP25, express a functional v-raf oncogene. However, they are non-tumorigenic, do not form colonies in soft agar and possess a flat morphology. CHP25 cells are resistant to re-transformation by sis, ras, tyrosine kinase- as well as serine/threonine kinase-encoding oncogenes suggesting that Raf functions downstream of most membrane associated signal transducers. In contrast to v-raf transformed cells, in which the endogenous Raf-1 protein kinase is constitutively activated, v-Raf in CHP25 cells does not activate endogenous Raf-1 kinase. Since mitogen regulation of Raf-1 kinase in CHP25 cells is intact, we conclude that CHP25 cells are blocked at the level of Raf-1 substrate phosphorylation. Consistent with this interpretation CHP25 cells show specific alterations of early gene induction. The serum induction of c-fos and junD as well as the serum and TPA (12-O-tetradecanoylphorbol-13-acetate) induction of junB and egr-1 are almost completely abolished. Only v-fos can transform CHP25, whereas c-fos, v myc, c-jun and junB are ineffective. These data suggest that the lesion responsible for the revertant phenotype of CHP25 cells is the inability to activate the AP-1 complex. We conclude that Raf-1 signaling is essential for transformation of NIH/3T3 cells by peripheral oncogenes and for regulation of a subset of early response genes by TPA and serum growth factors. PMID- 8426743 TI - Expression of the colony-stimulating factor 1 receptor in B lymphocytes. AB - The FMS proto-oncogene encodes for the colony-stimulating factor 1 receptor (CSF 1R), whose expression within the haematopoietic system has previously been thought to be restricted to cells of the mononuclear phagocyte lineage. We have studied the expression of the CSF-1R in peripheral blood mononuclear cells by indirect immunofluorescence and flow cytometry. FMS expression was detected on both monocytes and B lymphocytes from all samples analysed, including 14 haematologically normal individuals and 31 patients (23 in remission following cytotoxic therapy for lymphoma, six with B-cell chronic lymphocytic leukaemia and two with chronic myelomonocytic leukaemia). The level of FMS expression on B lymphocytes was lower than the level of expression detected on monocytes isolated from the same sample. FMS mRNA expression in B lymphocytes has been confirmed by a reverse transcription-polymerase chain reaction (RT-PCR)-based technique and Northern blot analysis. Thus, FMS may play a role in the normal function of B lymphocytes and, because of its potential oncogenic activity, may contribute to the pathogenesis of malignancies of this cell type. PMID- 8426744 TI - Functional domains of c-Myc involved in the commitment and differentiation of murine erythroleukemia cells. AB - In the early event of the induction of mouse erythroleukemia (MEL) cell differentiation, c-myc mRNA levels show a drastic change. The elevated expression of a transfected c-myc gene inhibits the commitment and differentiation of MEL cell transformants. In the present work, we have introduced human c-myc mutants into MEL cells under the inducible promoter to define the functional domains of c Myc involved in erythroid differentiation. The c-Myc domains necessary for commitment and differentiation are not co-localized; almost entire regions are required for inhibition of commitment, whereas domains II and IV that are essential for co-transforming activity with ras are required for inhibition of differentiation. Interestingly, mutants that delete domains for c-Myc dimerization motifs enhanced differentiation. These results suggest that c-Myc may regulate commitment and differentiation by interacting with proteins through different domains. PMID- 8426745 TI - Neoplastic transformation of a human keratinocyte cell line by the v-fos oncogene. AB - To analyze the role of the fos oncogene in the growth of human epithelial cells, we have transfected a non-tumorigenic human epidermal keratinocyte line (RHEK-1) immortalized by the Ad12-SV40 hybrid virus with a plasmid carrying the v-fos gene together with plasmid pSV2-neo which confers resistance to neomycin. Individual neomycin-resistant clones were isolated and characterized with respect to morphological alteration. Of 16 independent clones analyzed, two appeared morphologically transformed and formed foci in culture. Only the two clones with a transformed phenotype were found by Southern blot hybridization analysis to contain the transfected v-fos gene. These clones formed colonies in soft agar and induced tumors when transplanted into nude mice. Analysis of fos specific mRNA and protein demonstrated that the transfected v-fos gene was expressed in these two clonal lines. These findings suggest that expression of the v-fos gene might facilitate the process of neoplastic transformation of human epithelial cells in culture. This appears to represent the first demonstration of the transforming potential of the v-fos gene in human cells. PMID- 8426746 TI - Regulatory mechanism of 92 kDa type IV collagenase gene expression which is associated with invasiveness of tumor cells. AB - 92-kDa Type IV collagenase, a member of matrix metalloproteinases, is believed to play a critical role in physiological tissue-remodeling processes and also in many pathological conditions such as tumor invasion. We analyzed the 5'-flanking sequence of the 92 kDa type IV collagenase gene that controls the expression of the gene by ligating it to the chloramphenicol acetyltransferase gene. Deletion and mutation analysis revealed that three motifs, homologous to the binding sites for AP-1, NF-kappa B, and Sp-1 proteins, contributed positively to induction by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and tumor necrosis factor alpha (TNF alpha). The AP-1 site was indispensable but not sufficient for the induction and required synergistic cooperation with either the kappa B or the Sp-1 site. In OST cells, a nuclear factor which bound to Sp-1 was constitutively expressed, and those bound to AP-1 and kappa B elements were rapidly induced by TNF alpha treatment. Comparison of the findings with those for the promoters of other TPA inducible matrix metalloproteinases, interstitial collagenase and stromelysin 1, revealed that the signal to the AP-1 sites is common for the TPA-inducibility of the genes but that the signals to the kappa B or Sp-1 sites, which are not present in interstitial collagenase and stromelysin 1 promoters, are the unique determinant for the inducibility of the 92 kDa type IV collagenase gene. PMID- 8426747 TI - Co-purification of mitogen-activated protein kinases with phorbol ester-induced c Jun kinase activity in U937 leukaemic cells. AB - Phorbol esters, such as phorbol myristate acetate (PMA), cause differentiation of U937 human monomyelocytic cells along the macrophage pathway. Within 15 min of PMA treatment DNA binding of the c-jun transcription factor is increased and is accompanied by rapid changes in the phosphate content of the c-jun protein. Phorbol esters stimulate phosphorylation of serines 63 and 73 located within the A1 transactivation domain of c-Jun that have previously been shown to positively regulate activity. A protein kinase activity is detectable in extracts of phorbol ester-treated U937 cells that specifically targets these two serines. Using novel assays, the protein kinase activity has been purified over 1000-fold. The major portion of protein kinase activity co-chromatographs over three columns with pp42/44 mitogen-activated protein kinases as judged by immunological methods. The significance of these results with respect to mitogen-induced transcription of AP 1-responsive genes is discussed. PMID- 8426748 TI - Characterization of Ras effector mutant interactions with the NF1-GAP related domain. AB - The GTPase activating proteins Ras GAP and the neurofibromatosis-type 1 (NF1) gene product have been implicated as both potential mediators and regulators of the mitogenic effects of the ras proteins. In this study, the interactions of selected Ras effector mutants with the NF1-GAP related domain (NF1-GRD) were investigated. The NF1-GRD was unable to stimulate the GTPase of Ras[Asn33], Ras[Ser35] or Ras[Asn38], all transformation defective mutants. Each of these mutants had reduced but detectable binding to the NF1-GRD (apparent KD of 9 microM, 4 microM and 2 microM respectively, vs 0.5 microM for normal Ha-ras). The NF1-GRD was able to fully stimulate the intrinsic GTPase of the transformation defective Ras[Gly26Ile27] and Ras[Glu45] mutants, each of which bound the NF1-GRD with wild type affinity or better (KD = 0.13 microM and 0.4 microM respectively). The transforming Ras[Glu30Lys31] protein showed no GTPase stimulation and bound most poorly to the NF1-GRD (apparent KD of 16 microM). The interaction of the NF1 GRD with these specific Ras effector mutations is similar to that observed for Ras GAP. When the relative transforming activity of the valine 12 form of each Ras mutant was plotted against the apparent KD for NF1-GRD binding, little correlation was observed. These results support the hypothesis that the NF1 gene product does not function as a downstream effector of Ras in the mitogenic pathway. PMID- 8426749 TI - Transcriptional regulation in the testis: a role for transcription factor AP-1 complexes at various stages of spermatogenesis. AB - The products of two proto-oncogenes, c-fos and c-jun, have been implicated in signal transduction pathways as regulators of gene expression. Both proto oncogenes are members of gene families encoding closely related proteins that together make up transcription factor AP-1. The expression of members of this transcription factor has been associated with cellular pathways that result in both mitosis and differentiation. We have been studying the process of spermatogenesis, which is a complex, continual cycle of cell renewal, proliferation and differentiation. Using a seasonal breeder, the European red fox (Vulpes vulpes), as our model, we have examined the expression of five AP-1 family members (c-fos, fra-1, fra-2, c-jun and junB) with a view to elucidating their role in the regulation of spermatogenesis. Unique patterns of expression, falling into three broad categories, were observed for the five genes: (i) continuous expression throughout the spermatogenic cycle (c-fos); (ii) expression only at times corresponding to the onset and shutdown of spermatogenesis (fra-1, fra-2 and c-jun); and (iii) expression only at the onset of the cycle (junB). Furthermore, the proteins were expressed in both premeiotic and post-meiotic cell types, suggesting a role in haploid, as well as diploid, gene expression in this tissue. The data suggest distinct, although not necessarily unrelated, roles for the different components of transcription factor AP-1 in the regulation of spermatogenesis. PMID- 8426750 TI - Sequence and biological activity of chicken snoN cDNA clones. AB - cDNA clones of the ski-related gene, sno, were isolated from a chicken cDNA library and sequenced. In contrast to the human system, from which two forms of sno cDNAs have been isolated, we obtained only one type of chicken sno cDNA, that encoding snoN. The coding region for chicken snoN was inserted into the retroviral vectors RCAS(A) and RCASBP(A) and introduced into chicken embryo fibroblasts (CEFs) or quail embryo cells (QECs). Like the various forms of ski, snoN appears to be localized in the nucleus, and high levels of snoN expression cause transformation of CEFs and muscle differentiation of QECs. In contrast to ski however, low-level expression of snoN cannot induce transformation, and is only weakly myogenic. PMID- 8426751 TI - Gene rearrangement and truncated mRNA in cell lines with 11q23 translocation. AB - We have previously demonstrated that the breakpoints of t(11;19)(q23;p13) leukemias are within 360 kb of the CD3 gene. One of the phage clones, 6n, which was isolated from the yeast artificial chromosome clone yB22B2 containing CD3, was found to be within 60 kb of t(11;19) breakpoints. In this study, gene walking was conducted and two phage clones (lambda Hp8-3 and lambda Hp23-13) were isolated from a human placenta genomic library. Southern blot analysis with a genomic probe from lambda Hp8-3 detected gene rearrangements in t(4;11) and t(11;19) cell lines with BamHI digestion. Subsequently, using reiterated sequence free probes from both ends of 6n that detected transcriptional units in various hematopoietic cells, we isolated cDNA clones. These cDNA clones were classified into two groups (designated MLL-a and MLL-b), which do not hybridize to each other. Northern blot analysis with MLL-a cDNA detected 15-, 14- and 12-kb mRNAs, while MLL-b detected the additional 9.7- and 5-kb mRNAs in peripheral blood lymphocytes. MLL-b cDNA detected a truncated form of 12.5-kb mRNA in t(4;11) cell lines and a truncated form of 10-kb or 9.2-kb mRNA in t(11;19) cell lines. MLL-a did not demonstrate a truncated form of mRNA, but the stronger 14-kb signal was noted in t(4;11) cells, while this signal was very weak in t(11;19) cells. By Southern blot analysis, MLL-b cDNA detected gene rearrangement in cell lines with t(4;11) and t(11;19), whereas MLL-a did not. Furthermore, chimeric cDNA clones were isolated from cDNA libraries of t(4;11) and t(11;19) cell lines with a MLL-b cDNA probe. These results indicate that the MLL-b cDNA is derived from the common target gene involved in 11q23 translocation with 4q21 or 19p13. PMID- 8426752 TI - Alternative mRNA forms and open reading frames of the max gene. AB - The max gene encodes a heterodimeric partner of Myc. We have recently identified an alternative max mRNA (delta max) that contains an additional internal exon introducing an in-frame translational termination. Here we have studied the expression of human max mRNAs by Northern blotting analysis. In addition to the major 2.3-kb mRNA form, four bands were identified. Our results indicate that these bands represent differentially spliced mRNA forms, which contain altogether three open reading frames. In addition to the previously identified Max and delta Max proteins, sequence analysis of a 3.5-kb mRNA form predicted a protein that resembles delta Max in structure. Like delta Max, this protein enhanced the number of transformed foci in the ras-myc co-transformation assay. Although the 3.5-kb mRNA represents a minor form in actively proliferating cells, a shift from the major 2.3-kb mRNA to the 3.5-kb form was observed in response to high cell density or acidification of the growth medium. Our results indicate the presence of several differentially spliced mRNA forms of the max gene, and suggest a possible mechanism for the production of functionally distinct Max proteins. PMID- 8426753 TI - Low molecular weight GTP-binding proteins are altered in platelet hyperaggregation in IDDM. AB - We examined the hypothesis that hyperaggregating platelets from patients with insulin dependent diabetes mellitus (IDDM) have an alteration in location and function of the guanine nucleotide (GTP)-binding proteins. Platelets from 10 IDDM and 12 age-matched healthy control subjects were collected and washed. Thrombin induced platelet aggregation (0.025 and 0.05 units for 60 seconds) was increased in IDDM (8.3 +/- 1.8% vs 22.3 +/- 4.4%, P < .05 and 49.9 +/- 7.3% vs 70.9 +/- 7.0%, P < .05). Four small molecular weight GTP-binding proteins were identified by binding of [32P]-GTP on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the cytosol and membranes of these platelets. Each showed specificity for binding [32P]-GTP by competitive inhibition with unlabeled GTP. The total of the 27/28 kDa proteins was decreased in the membrane fraction (414 +/- 30 vs 252 +/- 40 dpm micrograms-1 protein x min, P < .05) and increased in the cytosolic fraction (62 +/- 8 vs 129 +/- 21 dpm unit-1 LDH x min, P < .05) in IDDM. The 21 kDa protein (60.3 +/- 3.5 vs 45.4 +/- 2.9 dpm micrograms-1 protein x min, P < .05) was decreased in platelet membrane in persons with IDDM. In conclusion, increased platelet aggregation in IDDM is accompanied by an altered cellular distribution of a 27/28 kDa GTP-binding protein. These data suggest that the low molecular weight GTP-binding proteins of the 27/28 kDa range may play an important regulatory role in the hyperaggregatory platelets in diabetes. PMID- 8426754 TI - Coding sequence of the overexpressed transcript of the putative oncogene PRAD1/cyclin D1 in two primary human tumors. AB - PRAD1 (cyclin D1) is a recently identified member of the family of cyclin genes, believed to play roles in regulating transitions through the cell cycle. The PRAD1 gene, located at 11q13, has been implicated in the pathogenesis of a variety of tumors, including parathyroid adenomas, t(11;14) bearing B-lymphoid tumors (particularly centrocytic lymphomas) where it is highly likely to be the BCL1 oncogene, and possibly in breast carcinomas and squamous cell cancers of the head and neck as well. PRAD1's tumorigenic influence appears to be effected through overexpression of its normal-sized transcript, but it has not been established whether the transcript's coding sequence is normal or contains oncogenic mutations. We have sequenced the coding region of the overexpressed PRAD1 transcript from two primary tumors with clonal PRAD1 region rearrangements: a benign parathyroid adenoma and a malignant centrocytic lymphoma. Each sequence is identical to the normal PRAD1 cDNA sequence, and presumably encodes normal PRAD1 protein. Thus, PRAD1 likely functions as a direct-acting oncogene whose rearrangement in tumors leads to overexpression or deregulated expression of its normal protein product. PMID- 8426755 TI - "Perceived satisfaction" counts in specialty choice. PMID- 8426756 TI - Medicare reform strikes again. PMID- 8426757 TI - 1991-92 in review: physicians, lobbyists work for good of all. AB - During the 1991-92 legislative session, several hundred health-related bills were introduced in the Senate and House of Representatives. Hard work by State Society physicians and lobbyists on these bills, addressing such issues as children's health insurance and certificate of need, will benefit all Pennsylvania physicians and their patients in the years to come. The Society's Division of Governmental Relations overviews the major issues and their outcomes. PMID- 8426758 TI - A summary: your society's health care reform proposals. Pennsylvania Medical Society Long Range Assessment Committee. AB - The State Society's Long Range Assessment Committee has been working for over a year to develop viable proposals for health care reform in Pennsylvania. Throughout the process, the Society has acknowledged that achieving consensus even among its members on many aspects of this complex issue will be problematic. However, the Society's primary goal in developing, disseminating, and getting input on these proposals is to make physicians' voices heard as the vital process of reforms begins. PMID- 8426759 TI - Dealing with physician-specific data reports: talking points for physicians. PMID- 8426760 TI - New secretary plans modernized health department. Interview by Maria T. Montesano. PMID- 8426761 TI - SAFE KIDS are no accident. PMID- 8426762 TI - AIDS survey shows need for more education. PMID- 8426763 TI - Staff training for practice in the '90s. The Health Care Group. PMID- 8426765 TI - A resolution: let's not lay blame for our health care dilemma. PMID- 8426764 TI - PRO ponders dilemma posed by "social factors". AB - Since the early years of Medicare, physicians have been reminded that Medicare is an insurance program, not a social program. Because social factors come into play in reviewing cases, the author finds himself wondering, "Is change needed? Would it help?" PMID- 8426766 TI - Reliability of observation variables in distinguishing infectious outcome of febrile young infants. AB - We prospectively evaluated 7 observation variables (level of activity, level of alertness, respiratory status/effort, peripheral perfusion, muscle tone, affect, feeding pattern) which qualify patient clinical appearance in order to determine reliability in distinguishing the infectious outcome of 233 febrile infants ages 0 to 8 weeks. Each variable was graded either 1, 3, or 5, with a higher score indicative of a greater degree of compromise. All infants received physical examination and sepsis evaluation (lumbar puncture, complete blood count/blood culture, urinalysis/urine culture). The 3 outcome groups compared were 29 cases of serious bacterial infections, (+SBI; 10 with bacterial meningitis, 12 with bacteremia, 7 with urinary tract infection), 45 cases of aseptic meningitis (AM) and 159 cases culture-negative with normal cerebrospinal fluid (CN-NCSF). The mean score for each of the 7 variables was significantly greater in the +SBI group compared with both the AM and CN-NCSF groups (P < 0.05), whereas there was no significant difference in mean score for each of the 7 variables between the AM and CN-NCSF groups. Stepwise discriminant analysis identified 3 variables that best distinguished outcome: affect; respiratory status/effort; and peripheral perfusion, which constituted the Young Infant Observation Scale. The mean total Young Infant Observation Scale score generated from assessing these 3 variables was significantly greater (P = 0.0001) in the +SBI, group (9) compared with both the AM (5) and CN-NCSF (5) groups. A total Young Infant Observation Scale score > or = 7 had a sensitivity of 76%, specificity of 75% and negative-predictive value of 96% for outcome of +SBI. PMID- 8426767 TI - Potential interventions for preventing pneumonia among young children: lack of effect of antibiotic treatment for upper respiratory infections. AB - Upper respiratory infections (URI) are a source of significant morbidity in childhood and have been associated with the development of certain bacterial infections. However, the high incidence of URI contrasted with the low incidence of lower respiratory infection (LRI) suggests a low rate of development of viral or bacterial LRI after URI. Because the etiology of URI is primarily viral, antibiotics do not have any significant effect on the URI episode itself but have been used to treat URI in hopes of preventing bacterial complications after URI. Meta-analysis of studies in developed and developing settings suggests that antibiotic treatment of children with URI does not shorten the course of URI and does not prevent the development of pneumonia. Several studies reporting both positive and negative results could not be included in the meta-analysis because they were not randomized trials or did not detail LRI outcomes in children sufficiently. Because of limitations in study design and definition of LRI, research in this area cannot be considered definitive. However, the weight of theoretical and experimental evidence is against antibiotic treatment of URI as a means of preventing the development of pneumonia after URI. PMID- 8426768 TI - Liver function of hepatitis B carriers in childhood. AB - This study followed 314 children who were carriers of hepatitis B virus for 2 to 4 years and compared them with noncarriers, matched for age and sex, from the same community. No confirmed carrier lost hepatitis B surface antigen. Hepatitis B e antigen (HBeAg) was lost at a rate of 10.6% per year; the rate of decay was not affected by age or gender. Liver enzymes were higher in HBeAg-positive than in HBeAg-negative carriers and loss of HBeAg was usually followed by return to values in the normal range. There was evidence, however, of persistent mild liver dysfunction in carriers even after development of antibody to HBeAg. Serum alanine aminotransferase concentrations above twice the upper limit of normal were observed in 7% of carriers on at least one occasion but persisted for more than 1 year in less than 1% and clinical manifestations were rare. The hepatitis B carrier state was uncomplicated during the course of this study. However, risks of subsequent serious disease in adult life may be significant and continued surveillance of carriers is important for individual protection and to determine adverse prognostic features. PMID- 8426769 TI - Combined active-passive immunization against the hepatitis B virus: five-year follow-up of children born to hepatitis B surface antigen-positive mothers. AB - In order to evaluate the duration of "protective" concentrations (i.e. > or = 10 IU/liter) of antibody to hepatitis B surface antigen (HBsAg) in children vaccinated against hepatitis B in infancy, we followed 146 children born to HBsAg positive mothers from birth to age 60 months. Children were seen at yearly intervals and tested for hepatitis B virus serologic markers. Of the children included in the study, 134 were protected against infection with development of antibody to HBsAg, 5 became HBsAg-positive and 6 failed to respond to the vaccine but did not become infected. Antibody concentrations fell progressively with the passage of time. The probability of maintaining a "protective" concentration of antibody in vaccine responders at age 60 months was 86% (95% confidence interval, 80 to 93%). Gender, ethnic origin, HBeAg status of the mother and immunization schedule had no influence on the rate of antibody loss. We conclude that in developed countries, the great majority of children vaccinated in infancy remain protected against infection at least until age 60 months. The need for booster doses of vaccine in this population will be determined by long term follow-up of immunized cohorts. PMID- 8426770 TI - Comparison of acellular pertussis vaccine with whole cell vaccine as a booster in children 15 to 18 months and 4 to 6 years of age. AB - The safety and immunogenicity of a booster dose of a new acellular pertussis component diphtheria-tetanus toxoids-pertussis vaccine (DTaP) were compared with whole cell pertussis component diphtheria-tetanus toxoids-pertussis vaccine (DTwP). Fifty children ages 15 to 18 months and 50 children ages 4 to 6 years were studied. The incidence of adverse reactions observed during the first 72 hours after vaccination in the DTaP/DTwP vaccinees were: pain, 32%/92% (P < 0.001); redness, 14%/24% (P = 0.2); swelling, 2%/14% (P < 0.03); fever, 52%/90% (P < 0.001); drowsiness, 14%/34% (P < 0.05); fussiness, 32%/88% (P < 0.001); and unusually poor appetite, 6%/42% (P < 0.001). The geometric mean titers of anti pertussis toxin and anti-filamentous hemagglutinin antibody were also significantly (P < 0.001) higher in the DTaP compared to the DTwP recipients. When administered as a booster dose this DTaP vaccine, which has been chosen by the NIH for the second pertussis vaccine clinical efficacy trial, was more immunogenic for pertussis toxin and filamentous hemagglutinin and caused fewer and less severe adverse reactions compared with the Connaught DTwP vaccine used in this study. PMID- 8426771 TI - Evaluation of oral erythromycin and local isoniazid instillation therapy in infants with Bacillus Calmette-Guerin lymphadenitis and abscesses. AB - A randomized placebo-controlled prospective trial was conducted to evaluate the efficacy of erythromycin therapy in 69 patients affected with Bacillus Calmette Guerin lymphadenitis. When patients who developed subsequent regional abscesses were excluded, erythromycin caused significantly earlier resolution of lymphadenitis (5.1 months vs. 5.7 months for placebo; P < 0.01) compared with placebo. There was no significant difference in the proportion of patients who developed subsequent regional abscesses between the 2 groups (47% for erythromycin, 60% for placebo, P = 0.14). When the entire group of 69 patients was evaluated for "duration to heal" (regardless of subsequent abscess formation), erythromycin therapy (4.1 +/- 1.5 SD months) did not differ significantly from the placebo group (3.5 +/- 1.3 months, P = not significant). Patients who developed subsequent abscess (n = 36) along with those with B. Calmette-Guerin regional abscesses at presentation (n = 27) were further studied to compare oral erythromycin therapy with that of single dose 50-mg intranodal isoniazid instillation. Local isoniazid therapy caused significantly earlier resolution of the abscesses (3.9 months) compared with erythromycin therapy (5.2 months; P < 0.001). PMID- 8426772 TI - Multiply resistant nontyphoidal Salmonella gastroenteritis in children. AB - From January, 1990, to December 31, 1990, 75 children with multiply resistant Salmonella gastroenteritis were studied at the Children's Hospital "Ricardo Gutierrez" of Buenos Aires. These children ranged from 1 month to 15 years of age. Infection was community-acquired in 20 (26.6%), nosocomially acquired in 50 (66.7%) and undetermined in 5. Thirty-nine (52%) had grossly bloody stools. Fever occurred at some point in the clinical course in 61 children (81.3%) with a duration of 1 to 33 days (mean, 6.7 days). The duration of diarrhea (1 to 69 days) was longer in those who developed complications (P < 0.001). Six (8%) developed enterocolitis (2 with bowel perforation), 1 had a pulmonary abscess and 8 (11.4%) had bacteremia; 4 children died (5.3%). Salmonella typhimurium was the most common serovar (85.3%). Ninety percent minimum inhibitory concentration studies demonstrated that all strains were resistant to ampicillin (> 128 micrograms/ml), cephalothin (> 128 micrograms/ml), cefuroxime (> 128 micrograms/ml), nalidixic acid (> 256 micrograms/ml), rifampin (> 256 micrograms/ml), gentamicin (> 256 micrograms/ml) and tobramycin (256 micrograms/ml); 77.3% of strains were resistant to ceftazidime (32 micrograms/ml), 97.6% to netilmicin (> 256 micrograms/ml), 92.8% to amikacin (256 micrograms/ml), 24.4% to isepamicin (32 micrograms/ml), 5.3% to chloramphenicol (4 micrograms/ml) and 2.7% to cefoxitin (2 micrograms/ml). The 90% minimum inhibitory concentration of cefotaxime and ceftazidime was reduced by the addition of clavulanate. Aggressive multiply resistant Salmonella strains are a major pediatric problem in Buenos Aires. PMID- 8426773 TI - The ringworm riddle: an outbreak of Microsporum canis in the nursery. AB - Tinea capitis, the most common fungal infection in children, is rare in neonates. We report six patients in a Level II intermediate care nursery who developed nosocomial dermatophyte infections caused by Microsporum canis. The investigation, which led to the identification of a nurse as the common source, is described. The nurse had an indolent infection with M. canis. Human to human transmission is exceedingly rare for M. canis. The literature regarding neonatal dermatophyte infections is discussed in relation to the reported outbreak. PMID- 8426774 TI - Neonatal Trichosporon beigelii infection: report of a cluster of cases in a neonatal intensive care unit. AB - Trichosporon beigelii, a ubiquitous yeast found in soil, causes superficial dermatologic infections in normal hosts and rare cases of disseminated disease among immunocompromised patients. Neonatal cases are exceptionally rare. We report a cluster of cases of T. beigelii infections in a tertiary care hospital in Rochester, NY, during May to July, 1991. Three cases occurred in very low birth weight premature infants (23 to 25 weeks of gestation), two of whom died. The organism was isolated from urine alone in one case, skin and blood in one case and blood, tracheal aspirate and central venous catheter tip in one case. In a fourth, full term infant with respiratory distress syndrome T. beigelii was grown only from a femoral central venous catheter tip with no clinical evidence of infection. An epidemiologic investigation was performed and the mode of transmission in this outbreak was not identified, although cross-infection was suspected in the initial two cases. Our isolates were inhibited but not killed by usually achievable concentrations of amphotericin B. T. beigelii may cause outbreaks of serious infection in neonatal intensive care units, especially among premature infants. PMID- 8426775 TI - Individualized pharmacokinetic profiles to compute vancomycin dosage and dosing interval in preterm infants. PMID- 8426776 TI - Lack of effect of clinical research protocol participation on morbidity measures in human immunodeficiency virus-infected children. PMID- 8426777 TI - Management of recalcitrant pain in a pediatric acquired immunodeficiency syndrome patient. PMID- 8426778 TI - Torticollis and bacterial meningitis. PMID- 8426779 TI - Successful use of extracorporeal membrane oxygenation in a newborn with herpes simplex virus pneumonia. PMID- 8426780 TI - Group B streptococcus as a cause of neonatal bullous skin lesions. PMID- 8426781 TI - Obturator internus abscess. PMID- 8426782 TI - Yersinia enterocolitica septicemia in infants younger than three months of age. PMID- 8426783 TI - Follow-up urine cultures in patients with acute pyelonephritis. PMID- 8426784 TI - Prolonged fever and diarrhea in a ten-year-old. PMID- 8426785 TI - C-reactive protein in febrile neutropenic patients. PMID- 8426786 TI - Sudden infant death among children born to women with human immunodeficiency virus type 1 infection. PMID- 8426787 TI - Diagnosis of urinary tract infection in a young infant. PMID- 8426788 TI - Risks of chickenpox in asthmatic children receiving inhalation steroids and therapeutic recommendations. PMID- 8426789 TI - Perestroika for the children. PMID- 8426790 TI - Working in UNISON. PMID- 8426791 TI - Dreams for the new year. PMID- 8426792 TI - Mentally disordered offenders. Widening the safety net. PMID- 8426793 TI - Mentally disordered offenders. Diverting tactics. PMID- 8426794 TI - Mentally disordered offenders. Who cares? PMID- 8426795 TI - A call for justice. PMID- 8426796 TI - Drama class. PMID- 8426797 TI - Making sense of ambulatory blood pressure monitoring. PMID- 8426798 TI - Gender and learning disabilities. Women's world. PMID- 8426799 TI - To believe or not to believe. PMID- 8426800 TI - Death on the operating table. PMID- 8426801 TI - Reaching out. PMID- 8426802 TI - Answer that soon. PMID- 8426803 TI - Nursing overseas. Making the most of things. PMID- 8426804 TI - Do patients know enough about epilepsy? PMID- 8426805 TI - Traditional versus 'causal' uniforms. PMID- 8426806 TI - Primary nursing. Intensive changes. PMID- 8426807 TI - Primary nursing. Team spirit. PMID- 8426808 TI - Testing for HIV in saliva works well. PMID- 8426809 TI - Troubled waters. PMID- 8426810 TI - Age of neglect? PMID- 8426811 TI - Caught unawares. PMID- 8426812 TI - Baby brainwave. PMID- 8426814 TI - All change--Community Care Act. PMID- 8426813 TI - Falling through the holes. PMID- 8426815 TI - Arrangements for care--Community Care Act. PMID- 8426816 TI - Out in force--Community Care Act. PMID- 8426817 TI - Children of the war. PMID- 8426818 TI - Abdominal massage. PMID- 8426820 TI - Nurse or potato peeler? PMID- 8426819 TI - Stress and the CPN (community psychiatric nurse). PMID- 8426821 TI - Positively French--nursing overseas. PMID- 8426822 TI - Special support. Mind your back. PMID- 8426823 TI - Emotional minefield. PMID- 8426824 TI - Return of the deadly disease. Health care overseas. PMID- 8426825 TI - Asthma--which peak flow meter? PMID- 8426827 TI - HIV patients slam insensitive care. PMID- 8426826 TI - Asthma--the insider's view. PMID- 8426828 TI - Growth and development. PMID- 8426829 TI - Obstetric medication versus residential area as perinatal risk factors for subsequent adult drug addiction in offspring. AB - In an attempt to explain pronounced uneven distributions of births of subsequent amphetamine and opiate addicts at seven hospitals in Stockholm, two possible mechanisms for adult drug addiction were weighed against each other: (1) risk factors associated with the obstetric care at the hospitals of birth of the addicts and (2) risk factors associated with the phenomenon of 'contagious' transmission of drug addiction in certain residential areas during adolescence. The subjects comprised 200 amphetamine addicts and 200 opiate addicts born between 1945 and 1966. By loglinear analysis the relative risk for future addiction was determined for eight residential areas as well as for the seven hospitals and four periods of birth. For the opiate addicts only one weak association was found for the residential area, which could not explain fully a clustering of births at any particular hospital. For the amphetamine addicts, hospital of birth was found to be an important risk factor even after controlling for residential area. Hence, the variable residential area has not been able to explain the uneven distribution of births of drug abusers among the studied hospitals. PMID- 8426830 TI - Social differences in late fetal death and infant mortality in Sweden 1985-86. AB - Social differences in late fetal death and infant mortality were examined in a population-based prospective study. All singletons born to Nordic citizens in Sweden 1985-86 were included, 185,156 births in all. The overall rates of late fetal death and infant mortality were 3.5 and 5.3 per 1000 respectively. Socio economic status of the household (SES) and mothers' education were used as social indicators. Logistic regression analyses showed significant odds ratios between 1.3 and 1.8 for late fetal death for blue-collar workers and women with less than 12 years education. The analyses of neonatal mortality showed a U-shaped relationship: both unskilled blue-collar workers and high level white-collar workers had significant odds ratios (OR) as compared with intermediate level white-collar workers (OR = 1.5). Similar results were obtained when using the mothers' education as indicator of social status: 9 years education or less or 15 years or more were associated with significant higher mortality rates than 12 years education (OR = 1.6). An inverse relationship between social status and post-neonatal mortality was seen in the crude analysis. Mothers' education revealed more social differences than SES (crude OR = 2.0 and 1.5 respectively in the least privileged group). However, when adjusting for the effects of maternal age, parity and smoking, no significant odds ratios for the social variables were obtained. PMID- 8426832 TI - Fumes from the spleen. PMID- 8426831 TI - Patterns of use of obstetrical interventions in 12 countries. AB - Recent obstetrical practice trends in 12 countries were surveyed. There was a 3 fold difference in caesarean section rates and a 10-fold difference in instrumental vaginal delivery rates among countries. There was a net increase in the caesarean section rate of all countries over the study period and a net decrease in the instrumental vaginal delivery rate of some countries. There was a decrease in the caesarean section rate during the last year of observation in Australia, Denmark and Finland. In general, countries with high caesarean rates also had high instrumental vaginal delivery rates. There was no consistent relationship between use of caesarean section and use of instrumental vaginal delivery, although in several countries increasing use of caesarean section was accompanied by decreasing use of instrumental vaginal delivery. Oxytocin use rates were associated positively with instrumental delivery but not with caesarean section rates. While it was not possible to determine the proportions of women who received appropriate obstetrical care, we can infer that a significant proportion of interventions were unnecessary or only marginally beneficial. Continued increases in rates of obstetrical intervention are unlikely to result in improvements in birth outcome overall and may pose a risk to mothers and their newborns. PMID- 8426833 TI - The associations between childhood asthma and atopy, and parental asthma, hay fever and smoking. AB - The aim of this analysis was to examine the degree to which a life time prevalence of asthma in a 7-year-old child is statistically associated with atopic conditions of the child, and with parental asthma, hay fever and smoking. In 1968, 8585 children who were born in 1961 and who were attending school in Tasmania were surveyed. This comprised 99% of the eligible population. The prevalence of a history of asthma in the 7-year-olds was 16.2% (males 19.0%, females 13.2%). Multiple logistic regression analysis showed that a history of asthma in a 7-year-old was associated with the child being male (odds ratio [OR] 1.56; 99% confidence interval 1.30-1.86), having a history of hay fever (3.86; 3.12-4.78), eczema (2.04; 1.63-2.55), hives (1.34; 1.09-1.65) or allergy to foods or medicines (1.70; 1.26-2.30), the child's mother or father having a history of asthma (2.63; 2.08-3.31 or 2.52; 1.99-3.19, respectively), and the mother being a smoker (1.26; 1.05-1.51). Parental hay fever and paternal smoking were not independently associated with childhood asthma. The strength of association between childhood asthma and parental asthma was independent of the sex of either the parent or the child, and of atopic conditions in the child. In the 133 children for whom both parents were asthmatic, 65 (49%) had a history of asthma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426834 TI - Maternal age and cognitive and behavioural outcomes in middle childhood. AB - The relationships between maternal age and childhood cognitive and behavioural outcomes were analysed in a birth cohort of New Zealand children studied until the age of 13 years. Increasing maternal age was associated with increasing scores on tests of cognitive ability and achievement and with decreasing reports of conduct problems. The correlations between maternal age and childhood cognitive/behavioural outcomes ranged from 0.11 to 0.21 with a median value of 0.18. These associations were adjusted statistically for the effects of maternal social background at the time of birth and for childhood history subsequent to birth. This analysis suggested that the poorer outcomes for children born to younger mothers reflected the cumulative effects of two sets of factors which placed these children at a disadvantage. Firstly, the offspring of younger mothers tended to be born into relatively poorly educated, socially disadvantaged families and secondly, these children were exposed to less nurturant and more changeable home environments. The net effects of differences in social background at birth and childrearing environments subsequent to birth were sufficient to explain most of the apparent correlation between maternal age and childhood cognitive/behavioural outcomes. PMID- 8426835 TI - Second generation follow-up of the Danish perinatal study women: study design and factors affecting response. AB - To study the mother-infant correlation of fetal growth and duration of pregnancy, women who were born as subjects in the Danish Perinatal Study (1959-61) were traced and interviewed, and the pregnancy and birth records of their children were abstracted. The study population consisted of 159 women who were small-for gestational age (SGA) at birth, 162 who were preterm, 38 who were both preterm and SGA, and 939 term, appropriately-grown control women. Methods for sample selection, measuring gestational age and fetal growth in both generations, locating and interviewing the women, abstracting the records of their children, and obtaining paternal birth and adult stature are described. A total of 84.5% of the selected women were successfully interviewed; the fraction interviewed did not differ by maternal birth status. The medical records of over 98% of pregnancies to the study women were abstracted, making it possible to study various factors associated with completion of an interview. By a variety of measures, women of higher socio-economic status were more likely to be interviewed. Birthweight and adult weights were available for 63 and 73% of the children's fathers. PMID- 8426836 TI - DNA sequence of the androgen receptor in prostatic tumor cell lines and tissue specimens assessed by means of the polymerase chain reaction. AB - Essentially all prostatic carcinomas relapse to an androgen-independent stage during androgen ablation therapy. The underlying genetic changes are still unclear. Such changes are suspected to affect the androgen-signalling pathway as well as growth promoting and inhibiting factors. This study was undertaken to test for structural changes of the androgen receptor in prostatic tumor cell lines and primary tumors. Complementary DNA (cDNA) fragments of the androgen receptor (AR) were isolated from the cell lines LNCaP, PC-3, and DU 145, ten tissue specimens obtained by radical prostatectomy, and five fine-needle biopsies by means of the polymerase chain reaction (PCR) technique. Fragments encoding the hormone- and DNA-binding domains were analyzed by DNA sequencing. The PCR technique is highly sensitive and especially recommended for the analysis of small tissue samples, such as those obtained by fine-needle aspiration. No alterations were detected in the tissue specimens and the five fine-needle aspirates. In the three tumor cell lines that represent late stages of prostatic tumor, different findings were obtained. The androgen-independent DU 145 cells did not express androgen receptors, whereas the PC-3 cells, which are also androgen-independent, expressed very low levels of normal AR. In contrast to this, the androgen-dependent LNCaP cells expressed high levels of structurally abnormal androgen receptors. These results suggest that androgen receptor mutations are probably uncommon molecular events in the early stages of prostatic cancer. Qualitative and quantitative changes, however, seem to occur in advanced prostatic cancer. PMID- 8426837 TI - Mutant p53 expression in prostate carcinoma. AB - The expression of the mutant p53 tumor suppressor gene was evaluated in 33 human prostate carcinomas. Using an immunohistochemical method with monoclonal antibodies PAb 1801 and PAb 240, 26 (79%) tumors demonstrated positive immunostaining for mutant p53. Only areas of glandular tumor were positive, with adjacent stromal elements and areas of glandular hyperplasia being negative. The predominant staining pattern was cytoplasmic. This pattern may be related to p53 binding to certain heat shock proteins (HSP 72/73), as a monoclonal antibody to these proteins demonstrated a cytoplasmic location as well. These results demonstrate that abnormal p53 expression is a frequent event in prostate cancer. PMID- 8426838 TI - High-affinity L-aspartate transporter in prostate epithelial cells that is regulated by testosterone. AB - The prostate gland produces and secretes extraordinarily high levels of citrate. Studies with rat ventral prostate (VP) have demonstrated that aspartate can serve as a four-carbon source of oxalacetate in the synthesis of citrate. To achieve this, prostate secretory epithelial cells must contain a transport system for the active uptake of aspartate from circulation. The present studies with VP epithelial cells confirm the existence of a Na(+)-dependent high-affinity L aspartate transporter. The transporter has an optimal pH approximately 7.5 and is temperature dependent. It appears to be an anionic amino acid transporter capable of transporting L-glutamate but not basic or neutral amino acids. The transporter is inhibited by ATPase inhibitors, thereby indicating its dependency on a Na+ gradient. The characteristics of the high-affinity L-aspartate transporter are consistent with its operation at the basilar membrane for the transport of circulating aspartate into the cell. Castration (24 hr) resulted in a significant decrease in the ability of VP epithelial cells to transport L-aspartate. The administration of testosterone to castrated rats completely restored L-aspartate transport. In addition, in vitro testosterone addition (10(-8) M for 30 min) to isolated prostate epithelial cells markedly increased L-aspartate transport. Both cycloheximide and actinomycin inhibited the testosterone effect. The studies reveal that testosterone is a regulator of this Na(+)-dependent high-affinity L aspartate transporter. The mechanism of this testosterone effect appears to involve both RNA and protein synthesis. We now have a model system to elucidate this novel effect of testosterone. PMID- 8426839 TI - Castration rapidly results in a major reduction in epithelial cell numbers in the rat prostate, but not in the highly differentiated Dunning R3327 prostatic adenocarcinoma. AB - Rats with implanted highly differentiated Dunning R3327PAP prostatic tumors were castrated, and at 3, 7, and 14 days thereafter, the effects on tumor volume, epithelial cell numbers, and sizes were quantified using morphometrical methods. The castration response on these parameters was also examined in the normal prostate of the same rats. Castration resulted in a rapid decrease in organ volume, epithelial cell number, and size in the normal prostate, and morphological signs of epithelial cell death (apoptosis) were observed. Tumor growth and mitotic index were reduced, but there were no signs of increased apoptosis, and cell numbers remained fairly constant in the Dunning tumors during the study period. It is concluded that the castration-induced inhibition of tumor growth is caused by factors other than tumor cell death. PMID- 8426840 TI - A detailed colostomy procedure and its application to quantify water and nitrogen balance and urine contribution to thermobalance in broilers exposed to thermoneutral and heat-distressed environments. AB - A surgical procedure for the separation of avian urinary and fecal excrement, in birds as young as 3.5 wk of age, was developed and used to quantify ambient temperature effects on urine production and composition. The colostomized broilers were used in two experiments to estimate urine production, osmolality, and contribution to water, nitrogen, and thermobalance of birds exposed to thermoneutral (24 C, TN) and heat-distressed (35 C, HD) environments. Urine production averaged over three 12-h periods was greater (P < .05), at 101 versus 51 mL/12 h per kilogram of BW and osmolality lower (P < .05) at 142 versus 220 mOsm/kg urine during HD and TN, respectively, in Experiment 1; and averaged over one 12-h period 82 versus 32 mL/12 h per kilogram of BW and 136 versus 208 mOsm/kg in Experiment 2 for HD and TN, respectively. In Experiment 1, urine production per milliliter of water consumed was increased (P < .05) by HD during Periods 2 and 3 even though in both experiments the values were similar during the first 12 h. Both urinary nitrogen and osmolar losses were increased (P < .05) by 50 and 89.2%, respectively, during HD, but without effect on fecal nitrogen. Evaporative cooling of broilers was increased (P < .05) by 154% during HD. These studies provide a well-defined colostomy procedure and suggest that broiler urine production and composition as well as overall thermobalance, water, and nitrogen balance are strongly impacted by acute HD. PMID- 8426841 TI - Effect of dietary full-fat flax seed with and without antioxidant on the fatty acid composition of major lipid classes of chicken meats. AB - Male broiler chicks (Hubbard x Hubbard) were raised to 6 wk on one of five test diets: corn-soybean meal control diet and 15% full-fat flax seed (FFS) with and without the antioxidants mixed tocopherol (T), canthaxanthin (C), or both (FFS, FFS + T, FFS + C, FFS + T + C). The effects of the antioxidants on broiler performance, carcass yield, and fatty acid composition of the white and dark meat lipid were investigated. Particular attention was paid to the n-3 fatty acid composition of triglyceride, phosphatidyl ethanolamine, and phosphatidyl choline fractions. The antioxidants influenced neither broiler performance nor yield. The birds did not utilize the experimental diets as efficiently as the corn-soybean meal diet. The fatty acid compositions of white and dark meat lipids of broilers were influenced by dietary fatty acids, but to different extents among the lipid fractions. Fatty acids with 20 or more carbon atoms were moderately distributed in the phosphatidyl choline fraction as compared with the triglyceride and phosphatidyl ethanolamine fractions. The presence or absence of antioxidants influenced the fatty acid composition and distribution in the phosphatidyl ethanolamine fraction of white meat. The white meat from birds fed FFS + T, FSS + C, and FFS + T + C diets had elevated levels of C18:3n3, C20:5n3, C22:5n3, and C22:6n3 and reduced levels of total saturates and n-6:n-3 as opposed to the FFS and corn-soybean meal groups. From a nutritional standpoint, the lower n-6:n-3 ratio, observed from birds fed the FFS diet seemed to be more desirable. PMID- 8426842 TI - An in vitro assay for prediction of broiler intestinal viscosity and growth when fed rye-based diets in the presence of exogenous enzymes. AB - A two-stage in vitro assay procedure was developed for predicting the efficacy of microbial enzyme sources in rye-based diets for broiler chicks. The procedure uses the complete diet as substrate to predict the in vivo intestinal viscosity and final weight of birds fed such diets. The optimum conditions for maximizing in vitro viscosity of a rye-based diet consisted of digesting .6 g of diet (ground through a 1-mm screen) in .9 mL of .1 N HCl containing 2,000 U pepsin/mL for 45 min at 40 C. At the end of this incubation phase, .3 mL 1 M NaHCO3 containing 8 mg pancreatin/mL (8 x USP) was added and the tube sealed and incubated for 2 h with intermittent vortexing. The digesta was then centrifuged (12,700 x g) and the supernatant collected for viscosity analysis using a digital viscometer. This assay was used for a rye-based diet containing either 0, .1, .2, .4, .8, or 1.6% of a xylanase source (experimental Trichoderma longibrachiatum product). The results were compared to weight and intestinal viscosity proximal and distal to Meckel's diverticulum of broilers (19 days of age) that had been fed these diets. The in vitro assay accurately predicted the in vivo intestinal viscosity (proximal r2 = .758, P < .0001, distal r2 = .667, P < .0001) and final weight of these birds (r2 = .660, P < .0001). The data suggest that the in vitro assay is a reliable assay for assessing the growth-promoting ability of an enzyme preparation in chicks fed rye-based diets. PMID- 8426843 TI - Vitamin profiles of eggs as indicators of nutritional status in the laying hen: vitamin A study. AB - Laying hens were fed deficient diets, unsupplemented or supplemented with one, two, or four times the National Research Council requirement level of vitamin A for 27 wk. Vitamin A levels of egg yolk from hens fed the unsupplemented diet slowly declined but levels of vitamin A stabilized at 12 wk in the supplemented groups and were related to dietary level. Egg production of hens fed the unsupplemented diet declined significantly after 12 wk and egg yolk levels continued to decline to Week 27. Hatchability of eggs laid by the unsupplemented hens was significantly lower at Weeks 25 and 26 than hatchability for those hens receiving dietary vitamin A. Hen weight, egg weight, and shell thickness were not consistently changed by treatment. In a second experiment, hens were fed a diet with and without 9,000 IU vitamin A/kg of diet. With birds receiving the supplemented diet, egg yolk vitamin A levels were stable after 13 wk, whereas egg yolk levels from unsupplemented hens declined slowly. Egg production with the unsupplemented diet was significantly reduced after 16 wk but the magnitude of the reduction was less than that in the first experiment, although egg vitamin A levels were as low as those in the first experiment. Hatchability of eggs during Weeks 25 to 28 was greatly reduced. Egg weight, albumen score, and chick weight were not consistently altered by vitamin A status. Because declines in egg yolk vitamin A do not precede declines in egg production they are not useful to predict future vitamin A deficiency in laying hens. PMID- 8426844 TI - Nutritional evaluation of copper-lysine and zinc-lysine complexes for chicks. AB - Three chick experiments were conducted to evaluate two mineral-Lys complexed products, Cu-Lys and Zn-Lys. Relative bioavailability of Lys, Cu, and Zn were assessed separately using diets specifically deficient in each of these nutrients and by employing criteria that respond in a straight-line fashion to supplementation with the nutrient in question. Common-intercept multiple linear regression (slope-ratio) of weight gain, bile Cu concentration, and total tibia Zn regressed on supplemental intake of Lys, Cu, and Zn, respectively, were used to assess bioavailability values relative to the standards (i.e., L-Lys.HCl, CuSO4.5H2O, and ZnSO4.H2O). Lysine bioavailability of Cu-Lys and Zn-Lys were estimated as 89 and 109%, respectively, which were not different (P > .05) from 100%. Relative bioavailability of Cu in Cu-Lys and Zn in Zn-Lys were estimated as 120 and 106%, respectively. These values were not different (P > .05) from the standards, i.e., CuSO4.5H2O for Cu and ZnSO4.H2O for Zn. PMID- 8426845 TI - Comparison of two multiple blood sampling regimens using an indwelling vascular access device for investigations of the hen's ovulatory cycle and calcium metabolism. AB - The objectives of the present study were to compare the effects of two regimens of serial blood sampling on the concentrations of hormones and ions during the ovulatory cycle of the domestic hen, and to examine the effectiveness of an indwelling vascular access device for repeated collection of blood samples. Single Comb White Leghorn hens were bled every 2 h over a period of 24 to 26 h, either from one oviposition to the next oviposition (OVIP-OVIP), or from 10 h prior to ovulation until the same time 24 h later (AFTN-AFTN). Whole blood was analyzed for ionized calcium concentration. Plasma was analyzed for total calcium, inorganic phosphorus, 1,25-dihydroxycholecalciferol, estradiol-17beta, and progesterone concentrations. The OVIP-OVIP regimen, using oviposition as a reference point, provided more accurate measurements of ionized calcium, inorganic phosphorus, and estradiol-17beta than did the AFTN-AFTN regimen. Either bleeding regimen was suitable for observing the patterns of 1,25 dihydroxycholecalciferol and progesterone concentrations. The decrease in bound calcium concentration observed with both regimens appeared to be an artifact of repeated blood sampling. The chance of a bird laying an egg following her second oviposition was lower following the OVIP-OVIP regimen than the AFTN-AFTN regimen. The vascular access device was a helpful tool in procuring multiple blood samples for measurement of ions and hormones during the ovulatory cycle of the domestic fowl. PMID- 8426846 TI - The turkey major histocompatibility complex: characterization by mixed lymphocyte, graft-versus-host splenomegaly, and skin graft reactions. AB - A turkey subline at the Ohio Agricultural Research and Development Center was developed by DNA typing of the MHC using a chicken MHC Class II probe, and it segregated for specific MHC genotypes. Histocompatibility was examined between turkeys of known MHC genotype using skin graft procedures, mixed lymphocyte reactions (MLR), and graft-versus-host reactions (GVHR). Skin grafts were exchanged among 3-wk-old turkeys and it was found that when birds shared DNA patterns (genotypes), the skin grafts were usually accepted. In contrast, skin grafts were always rejected when birds did not share the identical DNA pattern. Similarly, MLR only occurred when the lymphocytes were derived from birds that did not share the same DNA pattern. Lastly, GVHR were examined in embryos injected with either sire or dam blood. The GVHR in embryos was dependent on the parental MHC genotype. Four MHC haplotypes were identified in the turkey subline. The turkey MHC has been designated MhcMega-B, and each of the haplotypes, Mega B(1) through Mega-B(4). PMID- 8426847 TI - T lymphocyte reactivity to glutamic acid-alanine-tyrosine in vitro does not reflect antibody response in vivo. AB - Mechanisms responsible for the differences in humoral immune response to GAT (a random linear amino acid polymer) were investigated in a line of chickens consisting of four sublines homozygous for Ea-B (B1 or B19) and high or low antibody response to GAT (Ir-GATH or Ir-GATL). Previous research provided evidence of chromosomal recombination between the serologically determined regions of the MHC (encoded by B-F and B-G genes) and the gene or genes that control immune response to GAT, but immune response to GAT did not seem to be mediated through differences in B-L gene products. In the present study, proliferation of GAT-primed T lymphocytes indicated that reactivity in vitro was not associated with antibody levels produced in the animal. Cell surface markers were identified by flow cytometry. Lymphocytes from Ea-B19 chickens that were Ir GATL had a higher percentage of suppressor T (CD8)-positive cells than did lymphocytes from Ir-GATH chickens. The Ea-B1 chickens that were Ir-GATL had a higher percentage of CD4-positive lymphocytes than did chickens that were Ir GATH. This may indicate that low response to GAT in the Ea-B19 chickens, but not in Ea-B1 chickens, is mediated by CD8-positive cells. The ability of antigen presenting cells (APC) to process and present GAT to antigen-primed T lymphocytes was tested in vitro. Measurements of lymphocyte proliferation indicated that, within the Ea-B1 blood type, APC from Ir-GATL chickens produced higher (P < .05) stimulation of both GAT low- and GAT high-responder lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426848 TI - Gene complementation in biological crosses for humoral immune response to glutamic acid-alanine-tyrosine. AB - Level of humoral immune response to GAT has been associated with the MHC of chickens. Matings between two unrelated lines of chickens with low antibody response to GAT (G-B1 and S1 Line B19L) resulted in progeny that were higher responders to GAT challenge (P < .05) than either of the parental lines. Progeny of matings between two related sublines that are low responders to GAT (S1 Lines B19L and B1L) had antibody responses to GAT that were not higher than the parental lines. Progeny of the between-line cross were backcrossed to S1 B19L and G-B1 (B13) parental lines, as well as mated inter se. These matings produced F2 progeny whose GAT response was significantly associated (P < .05) with their MHC (Ea-B) type. The progeny were of three MHC types (B19B19, B19B13, and B13B13) that bound 66.6, 71.9, and 4.6%, respectively, of the GAT in a radioimmunoassay. The results from these matings suggest that MHC or MHC-linked genes, as well as genes not linked to the MHC, contribute to control of humoral immune response to GAT in the lines of chicken tested. PMID- 8426849 TI - Comparative development of Eimeria tenella in primary chick kidney cell cultures derived from coccidia-resistant and -susceptible chickens. AB - A sixfold difference in resistance to coccidia (Eimeria tenella) infection between a resistant and a susceptible line of Auburn White Leghorn chickens, derived by selective breeding, has been reported. The purpose of the following study was to determine whether the resistance or susceptibility phenomenon in the Auburn lines could be manifested in a homogeneous group of isolated host kidney cells that support E. tenella development in vitro but not normally in vivo. Propagation of the parasite in host cells in vitro eliminates humoral and cellular elements of immunity, and allows the study of host genetic influences at the cellular level. Differences in parasite development were examined between the two lines of cells in vitro after 48 and 96 h of incubation; time periods that reflect initial infection of the host cells by the parasite and the subsequent asexual development. Quantification of differences by liquid scintillation counting was based on parasite-specific incorporation of pyrimidines, specifically [3H]-uracil. The results supported previous findings that overall E. tenella development was significantly greater in the host cells from the susceptible line than in the cells from the resistant cultures at both time periods. PMID- 8426850 TI - Reduced plasma cholesterol and lipoprotein in laying hens without concomitant reduction of egg cholesterol in response to dietary sorbose. AB - Experiments were conducted to determine the effect of sorbose on feed consumption, egg production and size, and cholesterol metabolism of laying hens. In Experiment 1, 87-wk-old laying hens (10 per treatment) were fed diets containing 0, 10, or 20% sorbose for 4 wk. In a second experiment, 108-wk-old laying hens (eight per treatment) were fed a control diet, a diet with 10% added sorbose, or the control diet with intake restricted to the level of sorbose treated hens for 4 wk. Feed consumption and egg production were recorded daily. Plasma and egg cholesterol levels were determined at 0, 2, and 4 wk. Plasma and egg very low density lipoprotein (VLDL) concentrations were determined after 4 wk. Egg production, feed intake, and body weight gain were significantly reduced by dietary sorbose. Egg and yolk weight and percentage yolk decreased in response to sorbose. Sorbose significantly reduced plasma cholesterol and VLDL by approximately 50%, compared with the hens fed a control diet. Egg cholesterol concentration (milligrams per gram of yolk) was significantly increased, although the reduction in yolk size resulted in similar total egg cholesterol (milligrams per egg). Restricting feed intake of laying hens significantly lowered plasma cholesterol, but not to levels comparable to that of sorbose-treated hens. The data indicate that substantial reduction of plasma cholesterol and VLDL by dietary sorbose was not accompanied by reduced egg cholesterol. PMID- 8426851 TI - Net transfer and incorporation of yolk n-3 fatty acids into developing chick embryos. AB - The effect of egg yolk fatty acid composition on the uptake and utilization of essential n-6 and n-3 fatty acids by the developing chick embryo was studied. Eggs were enriched with n-9, n-3, or n-6 fatty acids by incorporating sunflower seed high in oleic acid (C18:1 n-9), flax seed rich in linolenic acid (C18:3 n 3), or sunflower seed high in linoleic acid (C18:2 n-6) into the laying hen diets. Fertile eggs were collected and incubated. The fatty acid composition of eggs and newly hatched chicks were compared. Feeding diets containing flax seed increased (P < .05) total n-3 fatty to 528.4 mg compared with 53.9 and 39.3 mg for eggs from hens fed diets with high oleic acid or regular sunflower seed, respectively. Levels of C18:2 n-6 and monounsaturated fatty acids were higher in eggs from hens fed diets containing regular or high oleic acid sunflower seeds. Dietary fat did not influence the total lipid content of the egg yolk or total lipids of chick tissues. The fatty acid composition of the hatched progeny was significantly altered by egg yolk lipids. However, the percentage incorporation of essential n-6 and n-3 fatty acids into the progeny increased when yolk sources of these fatty acids were low. The developing chick embryo appeared to preferentially take up docosahexaenoic acid and arachidonic acid from the yolk lipids. Evidence also suggests that conversion of C18:2 n-6 and C18:2 n-3 to longer chain n-3 or n-6 fatty acids occurs during the incubation period. PMID- 8426852 TI - Science and its discontented. PMID- 8426853 TI - Studies of the enteric nervous system in Alzheimer disease and other dementias of the elderly: enteric neurons in Alzheimer disease. AB - Studies of the myenteric (Auerbach) plexus of esophagus, stomach, small intestine, colon, and rectum, by microdissection and pointcount morphometry, for 18 patients with Alzheimer disease (AD), eight with other types of dementia of the elderly, non-demented elderly patients, and younger control patients, show a normal loss of enteric neurons and plexus mass with age, comparable to that reported by others for rats and guinea pigs. Values for patients with AD or other non-AD dementias did not differ from those for elderly controls. Enteric neurons in AD or the other dementias studied showed no definite stain with ALZ-50, a monoclonal antibody to a derivative of the microtubule-associated protein tau, which stains degenerating cerebral neurons in AD. Although the processes in the central nervous system in AD affect some neurons derived from the neural plate, the results of this study suggest that the enteric neurons, which are of neural crest origin, are not affected in AD. Enteric neurons, at least by the methods of this study, do not provide a useful peripheral marker for AD. PMID- 8426854 TI - H-ras-1 gene mutations in basal cell carcinoma: automated direct sequencing of clinical specimens. AB - Ras oncogenes are activated by point mutations occurring in codons 12, 13 or 61 and almost any base pair mutation occurring within the first two positions of any of these codons results in activation of the gene. Although ras point mutations have been reported to occur in several skin neoplasms including squamous carcinoma, keratoacanthoma and melanoma, their frequency of occurrence in basal cell carcinoma is not known. We examined basal cell carcinomas from 13 patients for activating mutations in the H-ras-1 gene by automated direct sequencing of polymerase chain reaction assay amplified targets. We amplified sequences in exon 1 that flanked codons 12 and 13 and amplified sequences in exon 2 that flanked codon 61. The PCR products were centrifuged and directly sequenced using antisense primers in an automated sequencer using fluorescent dideoxyterminators. One tumor was found to show an activating G to A transversion in codon 13 which code for aspartic acid instead of glycine. Although H-ras mutations may be found in some skin tumors, they are not frequent in basal cell carcinomas. From this study we believe that direct sequencing of clinical material is of value and has advantages over other techniques. Additional studies need to be undertaken to understand the true clinical significance of ras mutations in basal cell carcinomas when they occur. PMID- 8426855 TI - Metallothionein gene expression in bladder cancer exposed to cisplatin. AB - Some 68% of bladder tumors will respond to cisplatin-based chemotherapy but only 30% will have a durable response. Recent studies have suggested that the metallothionein (MT) gene may produce cisplatin drug resistance in cell lines. To determine the role of MT gene overexpression in human tumors resistant to cisplatin, we evaluated 19 bladder tumors, seven of which had been exposed to cisplatin, for MT mRNA expression. By Northern analysis, four of the seven tumors exposed to cisplatin overexpressed the MT gene compared to untreated tumors. Of the three treated tumors without MT overexpression, one was a relapse 4 yr after the last dose of cisplatin and the other two received only one dose of chemotherapy. MT gene overexpression was found in some tumors that had failed cisplatin chemotherapy and may be a mechanism for drug resistance in bladder cancer. PMID- 8426856 TI - Interdepartmental quality assurance using coded autopsy results. AB - A system of reporting discrepancies between antemortem and postmortem diagnoses was devised for use in interdepartmental quality assurance at a large community medical center. Between 1988 and 1991, 213 autopsies comprising 6% of deaths were coded using the system of Battle et al. (1) and distributed to the quality assurance committees of clinical departments. A major discrepancy was found in 11.7% (95% confidence interval; 6.8 to 18.4%) of adult cases. The discrepancy rate tended to increase with increasing patient age, but there was no significant effect due to patient sex, race, hospital of origin, clinical department of origin, whether the autopsy was restricted to examination of only part of the body, or whether the patient died in or out of the hospital. The possible uses of the quality assurance system and problems encountered with its implementation are discussed. PMID- 8426857 TI - Pathogenesis of pseudoxanthomatous salpingiosis. AB - The abnormal localized deposition of lipofuscin-laden macrophages in the lamina propria of the fallopian tube (pseudoxanthomatous salpingiosis) is reported in two women. In both cases there was evidence of longstanding endometriosis; necrotic pseudoxanthomatous nodules of the ovary were present. Histochemical and ultrastructural analysis of the pigment confirmed that it is lipofuscin (ceroid). It is proposed that pseudoxanthomatous salpingiosis develops after an episode of acute salpingitis during which actively bleeding ovarian endometriosis leaks blood into the lumen of the fallopian tube. PMID- 8426858 TI - Tryptophan-induced lung disease: an immunophenotypic, immunofluorescent, and electron microscopic study. AB - L-tryptophan (LT) has been implicated as a causal agent in the recently recognized eosinophilia-myalgia syndrome (EMS). Pulmonary complications occur in up to 60% of patients. Lung biopsies have shown chronic interstitial pneumonia, mild eosinophilia, vasculitis and perivasculitis, and hypertensive pulmonary arteriopathy. Open lung biopsies from two women who developed respiratory symptoms associated with LT EMS were studied with a panel of antibodies to lymphoid cells, by transmission electron microscopy and by direct immunofluorescence for immunoglobulin and complement. The majority of the cells infiltrating the interstitium and around vessels were T-cells, with a predominance of CD8+ cells. Numerous alveolar macrophages were also identified. Rare polyclonal B-cells were also present. Ultrastructural studies confirmed the presence of interstitial and perivascular lymphocytes as well as occasional eosinophils. The inflammatory cells were also present in vessel cells. Fibrointimal thickening was not observed in the sections studied ultrastructurally. Immunofluorescent staining for IgG, IgA, C3, C4, albumin, kappa, and lambda was negative. There was scattered staining for fibrinogen in alveolar spaces. The etiology of LT EMS is still under investigation, although a contaminant acting in conjunction with host factors is the favored hypothesis. The results of this study indicate that T cytotoxic/suppressor cells may be intimately involved in the pathogenesis of the lung injury. PMID- 8426859 TI - Ischemic fasciitis: a juxta-skeletal fibroblastic proliferation with a predilection for elderly patients. AB - We are reporting six examples of a distinctive reactive fibroblastic proliferation occurring predominantly in elderly bed-ridden or convalescent patients that presents as an ill-defined mass often overlying a bony prominence or in close apposition to the periosteum of bone. Unlike conventional fasciitis, the lesion achieves a relatively larger size and is characterized by a biphasic appearance with an outer fringe of mitotically active fibroblasts and capillaries circumscribing a central area of liquefactive and focally coagulative necrosis. Follow-up information in all patients indicated no evidence of aggressive behavior. We believe these lesions arise on an ischemic basis, similar to a decubitus ulcer, from long-standing or intermittent pressure on the soft tissue structures. Because of their large size and close proximity to bone, they are commonly mistaken for a sarcoma. PMID- 8426860 TI - Effects of hormone therapy on the endometrium. AB - Hormone therapy induces a variety of histologic changes in the endometrium. Histologic patterns encountered in the most commonly used hormonal regimens are described. Oral contraceptives are associated with inactive, atrophic, or pseudosecretory glands and edematous stroma, decidual reaction without spiral arterioles, and stromal granulocytes. A high-potency progesterone may induce marked stromal and vascular hyperplasia and stromal myomatous nodules. Ovulation induction therapy accelerates the maturation of the stroma and is often associated with a discrepancy between the glands showing early secretory changes and an edematous, decidualized stroma. Hormone replacement therapy may stimulate endometrial proliferation if estrogens are used alone and produce endometrial hyperplasia and neoplasia. When estrogen and progesterone regimens are used, a wide range of histologic pattern may be found in various combinations: proliferative and secretory endometrium, glandular and adenomatous hyperplasia, stromal hyperplasia and decidual transformation, glandular metaplasia, atrophic endometrium, and any of the above with endometrial atrophy. Progesterone therapy for endometrial hyperplasia and neoplasia is followed by secretory changes of the endometrium, mostly subnuclear vacuoles, decidual reaction, and sometime squamoid "morules." Secretory changes seen after progesterone therapy in the endometrium do not rule out residual carcinoma. For hormone therapy for breast carcinoma, tamoxifen acts as an antiestrogen on the breast but often acts as an estrogen agonist on the endometrium; tamoxifen therapy may be associated with endometrial hyperplasia, polyps, adenomyosis, adenomatous hyperplasia, and adenocarcinoma. PMID- 8426861 TI - Radiofrequency capacitive heaters: the effect of coupling medium resistivity on power absorption along a mouse leg. AB - A capacitive radiofrequency source in conjunction with a temperature-controlled electromagnetic coupling medium has the potential of delivering uniform heating distributions in a mouse leg for experimental studies to investigate the use of hyperthermia as a treatment for cancer. The system has been adopted by a number of groups who have confirmed that uniform temperatures can be achieved in the presence of blood flow along a one-dimensional line which extends between the plates across the leg. In this paper, a simple mathematical model is presented and verified experimentally to demonstrate that parallel-plate capacitive radiofrequency heaters produce an inherent absorbed power distribution along the leg which is determined by the impedances across the loads. Hence, the thicker or thinner regions of the leg can be preferentially heated by using a coupling medium with lower or higher salinity, respectively. Uniform power absorption along the mouse leg required that the coupling medium had equivalent electrical properties to those of tissue. PMID- 8426862 TI - An analysis of the single-pool urea kinetic model and estimation of errors. AB - The single-pool urea kinetic model assumes that urea is distributed within the body in a volume Vf at a uniform concentration. It may be used to describe the clearance of urea during and following haemodialysis, and to determine the value of the urea concentration at the end of dialysis Cf or the urea generation rate G, and the urea distribution volume Vf. The protein catabolic rate (PCR) is obtained using the ratio G/Vf. The sensitivity of the predictions of the model to small errors in the experimentally estimated model parameters is evaluated and suggests that the model prediction of Cf is relatively insensitive to errors in the estimate of G, but more sensitive to errors in Vf and dialyser clearance K. The determination of G from urea concentration measurements made during dialysis alone is very sensitive to errors. The accurate estimation of G requires the use of concentration measurements made between dialyses. The calculation of the PCR is very sensitive to errors in G and Vf since these are not independent and must be used with caution, particularly when used to compare values between patients. PMID- 8426863 TI - Target apex-seeking in factor analysis of medical image sequences. AB - The aim of factor analysis of medical image sequences (FAMIS) is to estimate a limited number of physical or physiological fundamental functions. Its oblique rotation stage strongly affects the quality and the interpretation of the resulting estimates (factors and factor images). A new target apex-seeking method which integrates physical or physiological knowledge in this stage is described. This knowledge concerns some of the fundamental functions and reacts on the determination of all the factors. A simulated spectral study illustrates the method. We discuss its properties in comparison with the other approaches using a priori physical or physiological information. PMID- 8426864 TI - Geometrical reconstruction of images obtained with electronic endoscopy. AB - We use an electronic video endoscope (Fujinon EG7-HR2) to image the mitral valve in situ in an isolated pig heart preparation. From video recordings, images are digitized and analysed with a computer. A complication encountered during the study concerned the geometric wide angle distortion caused by the device. The present paper describes a method developed to reconstruct the image, and to correct for this distortion. In order to quantify the relation between object and image, model equations were formulated based on mild assumptions. Points in object space are transformed through a non-linear relation to corresponding points in image space. Furthermore, an oblique camera view, and an aspect ratio correction factor, are accounted for. As a test object, a regular grid of points was recorded at several distances from the camera. The images of the grid points were digitized, and the model equations were fitted to these data. From test measurements, carried out in air as well as in water, it was concluded that the distortion could be quantified by three parameters. Application of the straightforward correction procedure enables us to obtain quantitative information from endoscopic images. PMID- 8426865 TI - Investigation of the tissue equivalence of gels used for NMR dosimetry. AB - The transition of Fe2+ to Fe3+ in Fricke solution after irradiation results in a change of NMR proton relaxation times in agarose gels which can be used for the dosimetry of ionizing radiation. The main advantage of this system is the possibility of observing dose distributions in 3 dimensions in a medium which is supposed to be tissue equivalent. The aim of the present study was to quantify parameters which determine the tissue equivalence of NMR dosimetry gels. Electron densities and effective atomic numbers were calculated for gels with varying iron, sulphur and agarose concentration. The Hounsfield CT numbers so derived agree well with the CT numbers measured on a clinical CT scanner (effective photon energy 70.7 keV). The Hounsfield CT number of 7.2 +/- 1.5 (n = 9) measured for a 1.5% agarose gel doped with 0.5 mM ammonium ferrous sulphate and 125 mM sulphuric acid compares well with the calculated one of 8 +/- 5. Relaxation times were measured from a series of MR images obtained on a 1.5 T clinical MR scanner. The observed change in 1/T1 of the gel with dose was found to be linear up to 10 Gy (0.084 s-1 Gy-1). No difference in dose response for 10 Gy delivered by four different superficial radiation qualities (HVT = 1.4-7.5 mm Al) could be observed. These findings and the calculated effective atomic number of 7.46 demonstrate the close tissue equivalence of this agarose gel which makes it an ideal tool for the investigation of low energy therapeutic x-rays. PMID- 8426866 TI - A feasibility study of in vivo electromagnetic imaging. AB - Over the last decade several non-invasive electromagnetic techniques to determine human body composition have been described, based primarily on the good discrimination in electrical conductivity between fat tissue and water-bearing fat-free tissue. As an extension of such work, carried out within the Swansea In Vivo Analysis Research Group, an electromagnetic system is described which has the potential to generate useful in vivo images of body structure. Preliminary experiments on this electromagnetic scanning imager, using simple saline phantoms as objects, are discussed. It is shown that objects with electrical conductivities corresponding to fat and fat-free tissue can be differentiated and that the internal and external geometry of simple objects can be sensed. Images are presented which have been generated by several image reconstruction techniques. PMID- 8426867 TI - In vivo measurement of total body chlorine using the 8.57 MeV prompt de excitation following thermal neutron capture. AB - Prompt gamma neutron activation analysis with 238Pu/Be sources is used to measure total body chlorine (TBCl) in vivo following the reaction 35Cl(n, gamma)36Cl. The chlorine de-excitation at 8.57 MeV is used for calibration of the system for TBCl. Body hydrogen is used as an internal standard and TBCl is derived from the gamma-ray counts ratio of chlorine to hydrogen. The precision of the method, determined from replicate scans of a Bush-type phantom, is 4.9% (CV). To assess accuracy an anthropomorphic phantom consisting of minced meat was constructed. Replicate scans of this phantom yielded a mean total chlorine which was not significantly different from the chemical analysis value. The subject dose equivalent for the activation measurement is less than 0.3 mSv. Mean TBCl values for 63 male and 107 female healthy volunteers were in broad agreement with predicted amounts based on multiple regression equations developed at other centres from measurements using the delayed gamma approach. Good agreement was observed in 76 volunteers between total body water (TBW) measured by tritium dilution, after correction for non-aqueous hydrogen exchange, and TBW derived from the sum of extracellular water and intracellular water as measured by TBCl and total body potassium (TBK), respectively. PMID- 8426868 TI - Ion recombination corrections for plane-parallel and thimble chambers in electron and photon radiation. AB - The aim of this work was to investigate the collection efficiency of several popular ionization chambers used in high-energy electron and photon dosimetry. The recombination effect was evaluated in plane-parallel type ion chambers (Markus, NACP, Calcam and Vinten-631) and in a thimble-type chamber (NE2571). The chambers' response was studied in a pulsed accelerator beam, for both photons and electrons, and in continuous radiation from a 60Co machine. The study involved both theoretical and experimental evaluation of the recombination effect utilizing the methods described in the literature. The main conclusions to be drawn from this work are as follows: (i) For the Farmer NE2571 0.6 cm3 chamber used here, the correction factors required are of the order of 0.1% for 60Co radiation, 0.5% for 5 and 8 MV x-rays and 10 and 12 MeV electrons and 0.9% for 15 MV x-rays and 15 and 18 MeV electrons, all at typical clinical dose rates of 200 400 cGy min-1 (0.017 cGy pulse-1). (ii) The recombination effect for the plane parallel chambers was found to be smaller than for the thimble chamber, with values of < 0.1% in continuous radiation and 0.2-0.4% in pulsed radiation. PMID- 8426869 TI - The convergence of object dependent resolution in maximum likelihood based tomographic image reconstruction. AB - Study of the maximum likelihood by EM algorithm (ML) with a reconstruction kernel equal to the intrinsic detector resolution and sieve regularization has demonstrated that any image improvements over filtered backprojection (FBP) are a function of image resolution. Comparing different reconstruction algorithms potentially requires measuring and matching the image resolution. Since there are no standard methods for describing the resolution of images from a nonlinear algorithm such as ML, we have defined measures of effective local Gaussian resolution (ELGR) and effective global Gaussian resolution (EGGR) and examined their behaviour in FBP images and in ML images using two different measurement techniques. For FBP these two resolution measures are equal and exhibit the standard convolution behaviour of linear systems. For ML, the FWHM of the ELGR monotonically increased with decreasing Gaussian object size due to slower convergence rates for smaller objects. For the simple simulated phantom used, this resolution dependence is independent of object position. With increasing object size, number of iterations and sieve size the object size dependence of the ELGR decreased. The FWHM of the EGGR converged after approximately 200 iterations, masking the fact that the ELGR for small objects was far from convergence. When FBP is compared to a nonlinear algorithm such as ML, it is recommended that at least the EGGR be matched; for ML this requires more than the number of iterations (e.g., < 100) that are typically run to minimize the mean square error or to satisfy a feasibility or similar stopping criterion. For many tasks, matching the EGGR of ML to FBP images may be insufficient and >> 200 iterations may be needed, particularly for small objects in the ML image because their ELGR has not yet converged. PMID- 8426870 TI - SNR and noise measurements for medical imaging: I. A practical approach based on statistical decision theory. AB - A method of measuring the image quality of medical imaging equipment is considered within the framework of statistical decision theory. In this approach, images are regarded as random vectors and image quality is defined in the context of the image information available for performing a specified detection or discrimination task. The approach provides a means of measuring image quality, as related to the detection of an image detail of interest, without reference to the actual physical mechanisms involved in image formation and without separate measurements of signal transfer characteristics or image noise. The measurement does not, however, consider deterministic errors in the image; they need a separate evaluation for imaging modalities where they are of concern. The detectability of an image detail can be expressed in terms of the ideal observer's signal-to-noise ratio (SNR) at the decision level. Often a good approximation to this SNR can be obtained by employing sub-optimal observers, whose performance correlates well with the performance of human observers as well. In this paper the measurement of SNR is based on implementing algorithmic realizations of specified observers and analysing their responses while actually performing a specified detection task of interest. Three observers are considered: the ideal prewhitening matched filter, the non-prewhitening matched filter, and the DC-suppressing non-prewhitening matched filter. The construction of the ideal observer requires an impractical amount of data and computing, except for the most simple imaging situations. Therefore, the utilization of sub optimal observers is advised and their performance in detecting a specified signal is discussed. Measurement of noise and SNR has been extended to include temporally varying images and dynamic imaging systems. PMID- 8426871 TI - Dose conversion and wall correction factors for Fricke dosimetry in high-energy photon beams: analytical model and Monte Carlo calculations. AB - This paper presents the dose conversion and wall correction factors for Fricke dosimetry in high-energy photon beams calculated using both an analytical general cavity model and Monte Carlo techniques. The conversion factor is calculated as the ratio of the absorbed dose in water to that in the Fricke dosimeter solution with a water-walled vessel. The wall correction factor accounts for the change in the absorbed dose to the dosimeter solution caused by the inhomogeneous dosimeter wall material. A usercode based on the EGS4 Monte Carlo system, with the application of a correlated sampling variance reduction technique, has been employed in the calculations of these factors and the parameters used in the cavity model. Good agreement has been achieved between the predictions of the model and that obtained by direct Monte Carlo simulation and also with other workers' experiments. It is shown that Fricke dosimeters in common use cannot be considered to be 'large' detectors and therefore 'general cavity theory' should be applied in converting the dose to water. It is confirmed that plastic dosimeter vessels have a negligible wall effect. The wall correction factor for a 1 mm thick Pyrex-walled vessel varies with incident photon energy from 1.001 +/- 0.001 for a 60Co beam to 0.983 +/- 0.001 for a 24 MV (TPR(10)20 = 0.80) photon beam. This implies that previous Fricke measurements with glass-walled vessels should be re-evaluated. PMID- 8426872 TI - Prediction of controlled drinking by alcoholics and problem drinkers. AB - Recent research was reviewed on the following factors as predictors of controlled drinking (CD) by alcoholics and problem drinkers: severity of dependence, client attitudes and beliefs about controlled drinking and abstinence, previous treatment, pretreatment drinking style, psychological and social stability, demographic characteristics, family history of drinking, referral source, and posttreatment adjustment and drinking. No single personal characteristic has been consistently predictive, but there is convincing evidence that a lower severity of dependence and a persuasion that CD is possible are associated with CD after treatment. Prediction of CD might be improved by an approach that assesses the influence of three types of variables: enduring personal characteristics, changeable social and psychological characteristics, and transient precipitating events. PMID- 8426873 TI - Sex bias in the naming of stimulus persons. AB - Researchers often use sex-typed names (e.g., John vs. Joan) to identify stimulus persons' sex, assuming that such names communicate sex only. In fact, however, such names also create impressions that have little or nothing to do with sex. Study 1 analyzed the age connotations, intellectual-competence connotations, and attractiveness of sex-typed names used in 230 published studies on sexism and fear of success. On each of these variables, the literature was pervasively confounded in a manner favoring male stimulus persons. Study 2 found that the name biases reported in Study 1 were positively correlated with outcome measures in a sample of sexism studies, but only when names were presented with limited other information. Possible causes of the bias are discussed, and recommendations for naming stimulus persons are presented, including a list of male names and female names matched on several key variables. PMID- 8426874 TI - Impact of sexual abuse on children: a review and synthesis of recent empirical studies. AB - A review of 45 studies clearly demonstrated that sexually abused children had more symptoms than nonabused children, with abuse accounting for 15-45% of the variance. Fears, posttraumatic stress disorder, behavior problems, sexualized behaviors, and poor self-esteem occurred most frequently among a long list of symptoms noted, but no one symptom characterized a majority of sexually abused children. Some symptoms were specific to certain ages, and approximately one third of victims had no symptoms. Penetration, the duration and frequency of the abuse, force, the relationship of the perpetrator to the child, and maternal support affected the degree of symptomatology. About two thirds of the victimized children showed recovery during the first 12-18 months. The findings suggest the absence of any specific syndrome in children who have been sexually abused and no single traumatizing process. PMID- 8426875 TI - Psychopathology and early experience: a reappraisal of retrospective reports. AB - Three potential sources of error in retrospective reports of childhood experiences are documented: low reliability and validity of autobiographical memory in general, the presence of general memory impairment associated with psychopathology, and the presence of specific mood-congruent memory biases associated with psychopathology. The evidence reviewed suggests that claims concerning the general unreliability of retrospective reports are exaggerated and that there is little reason to link psychiatric status with less reliable or less valid recall of early experiences. Nevertheless, it is clear that steps must be taken to overcome the limitations of retrospective reports and enhance their reliability. PMID- 8426876 TI - Children's peer relations: a meta-analytic review of popular, rejected, neglected, controversial, and average sociometric status. AB - Two-dimensional sociometric models have had a critical role in the investigation of children's peer relations in the past decade. In a meta-analysis, fitting categorical models (Hedges, 1982), sociometric group differences on behavioral and information source typologies were assessed. The broad-band behavioral analysis showed that popular children's array of competencies makes them likely recipients of positive peer nominations, whereas high levels of aggression and withdrawal and low levels of sociability and cognitive abilities are associated with rejected peer status. A consistent profile marked by less sociability and aggression emerged for neglected status. Controversial children had higher aggressive behavior than rejected children but compensated for it with significantly better cognitive and social abilities. The moderator effects of narrow-band behavioral categories and information source were also examined. PMID- 8426877 TI - Definition and assessment of accuracy in social stereotypes. AB - A perennial issue in the study of social stereotypes concerns their accuracy. Yet, there is no clear concept of the various ways in which stereotypes may be accurate or inaccurate and how one would assess their accuracy. This article is designed to rectify this situation. Three forms of stereotype inaccuracy are identified: stereotypic inaccuracy, valence inaccuracy, and dispersion inaccuracy. The implications of each form are discussed, along with how each can be assessed using a full-accuracy design. Past research that has attempted to examine stereotype accuracy is reviewed, and new data on the issue are presented. Although of perennial interest, the theoretical and methodological difficulties of assessing stereotype accuracy are substantial. The goal in this article is to alert the researcher to these difficulties and point toward their solution. PMID- 8426878 TI - The importance of heritability in psychological research: the case of attitudes. AB - It is argued that differences in response heritability may have important implications for the testing of general psychological theories, that is, responses that differ in heritability may function differently. For example, attitudes higher in heritability are shown to be responded to more quickly, to be more resistant to change, and to be more consequential in the attitude similarity attraction relationship. The substantive results are interpreted in terms of attitude strength and niche building. More generally, the implications of heritability for the generality and typicality of treatment effects are also discussed. PMID- 8426879 TI - On variability, Simpson's paradox, and the relation between recognition and recall: reply to Tulving and Flexser. AB - Tulving and Flexser's (1992) defense of the Tulving-Wiseman law rests on the partitioning of data points into 2 sets, which they call constrained and unconstrained. This dichotomy depends crucially on the implicit assumption that within-condition variance is 0. Simulations are done to show the effects of variability on the maximum contingency that can be displayed by an average 2 x 2 table. The results help explain the form of the regularity known as the Tulving Wiseman law, as well as the conditions under which exceptions are found. This analysis reinforces the conclusion that the law is an artifact and serves as a reminder of the dangers posed by variability and Simpson's paradox when contingency analyses are done. PMID- 8426880 TI - Novelty monitoring, metacognition, and control in a composite holographic associative recall model: implications for Korsakoff amnesia. AB - This article stems from a technical problem in composite-trace distributed models of human memory and particularly in the Composite Holographic Associative Recall Memory (CHARM) model. Briefly, the composite trace--used as a central construct in such models--can become catastrophically out of control. To solve the problem, a prestorage novelty-familiarity monitor and a simple control procedure need to be implemented. Eight lines of experimental evidence converge on the idea that output from such a novelty-familiarity monitor underlies people's metacognitive judgments of feeling of knowing. Breakdown of the monitoring-control mechanism produces Korsakoff-like symptoms in the model. Impairments in feeling-of-knowing judgments and the failure to release from proactive inhibition, both characteristic of Korsakoff amnesia, are thus attributed to a monitoring-control failure rather than to deficits in the basic memory system. PMID- 8426881 TI - Memory independence and memory interference in cognitive development. AB - Recent experiments have established the surprising fact that age improvements in reasoning are often dissociated from improvements in memory for determinative informational inputs. Fuzzy-trace theory explains this memory-independence effect on the grounds that reasoning operations do not directly access verbatim traces of critical background information but, rather, process gist that was retrieved and edited in parallel with the encoding of such information. This explanation also envisions 2 ways in which children's memory and reasoning might be mutually interfering: (a) memory-to-reasoning interference, a tendency to process verbatim traces of background inputs on both memory probes and reasoning problems that simultaneously improves memory performance and impairs reasoning, and (b) reasoning-to-memory interference, a tendency for reasoning activities that produce problem solutions to erase or reduce the distinctiveness of verbatim traces of background inputs. Both forms of interference were detected in studies of children's story inferences. PMID- 8426882 TI - Four systems for emotion activation: cognitive and noncognitive processes. AB - The significant role of emotions in evolution and adaptation suggests that there must be more than 1 mechanism for generating them. Nevertheless, much of current emotion theory focuses on cognitive processes (appraisal, attribution, and construal) as the sole, or primary, means of eliciting emotions. As an alternative to this position, the present model describes 4 types of emotion activating systems, 3 of which involve noncognitive information processing. From an evolutionary-developmental perspective, the systems maybe viewed as a loosely organized hierarchical arrangement, with neural systems, the simplest and most rapid, at the base and cognitive systems, the most complex and versatile, at the top. The emotion-activating systems operate under a number of constraints, including genetically influenced individual differences. The hierarchical organization of the systems for generating emotions provides an adaptive advantage. PMID- 8426883 TI - Autognosis rounds for medical house staff. AB - The authors describe the autognosis countertransference rounds for medical house staff at the Massachusetts General Hospital. At these rounds, which have been held weekly for more than a decade in the intensive care unit, countertransference phenomena and their relationship to medical practice are discussed. Methods that have facilitated the autognostic process are provided and highlighted by brief case examples. Participants at these rounds report that their self-awareness increases and the clinical care they provide often improves. PMID- 8426884 TI - Cyclosporine psychosis. PMID- 8426886 TI - "On organicity" and "dementias". PMID- 8426885 TI - Catatonia responsive to intravenous lorazepam in a patient with cyclosporine neurotoxicity and hypomagnesemia. PMID- 8426887 TI - Psychological factors affecting physical condition. Endocrine disease literature review. AB - Research exploring the relationship between psychological factors and the onset, exacerbation, and perpetuation of endocrine diseases has focused primarily on three diseases: diabetes mellitus (DM), Graves' disease, and Cushing's disease. There is insufficient evidence to support the position that psychological factors directly affect the onset of DM. Recent laboratory studies suggest that stress is associated with changes in glucose regulation in a subset of diabetic patients and that temperament and coping strategies influence glycemic control in diabetic children and adolescents. Relaxation training may improve blood glucose control in non-insulin-dependent DM patients. There is no good evidence that psychological characteristics affect the development and course of thyroid disorder or Cushing's disease. Recommendations are made for future research. PMID- 8426889 TI - Incidence of suicidality in AIDS and HIV-positive patients attending a psychiatry outpatient program. AB - Current suicidality and many other characteristics were assessed in 183-patients in the various stages of HIV infection who were referred to a specialized HIV related psychiatric clinic. Intravenous drug use was the most common HIV risk factor. Patients with AIDS had significantly less current suicidal ideation than patients with AIDS-related complex and asymptomatic HIV positivity. The lower suicidality in AIDS patients was independent of age, gender, HIV risk group, and source of referral. Possible explanations of this association include denial, refocusing of life goals in AIDS patients, and psychological changes related to central nervous system impairment. PMID- 8426888 TI - Neurobiological models of obsessive-compulsive disorder. AB - The authors review current neurobiological models of obsessive-compulsive disorder (OCD). The rationale for anatomically based models is outlined and selected brain regions of interest are discussed. The authors conclude that there is abundant evidence to implicate multiple brain regions as sites of abnormality in OCD. A review of neuropharmacological concepts related to the serotonergic (5 HT) hypothesis of OCD is also presented. The authors conclude that anti obsessional agents may well have therapeutic effects via the 5-HT system, yet there is little unequivocal evidence to suggest an underlying abnormality in the 5-HT system. A comprehensive neurobiological model of OCD must acknowledge the interrelationship between multiple transmitter systems and integrate anatomical with neuropharmacological concepts. PMID- 8426890 TI - Cognitive and social correlates of the Toronto Alexithymia Scale. AB - The authors examine the relationship of the Toronto Alexithymia Scale (TAS) to sociodemographic characteristics, medical history, symptomatology, and illness cognition in a sample of 244 family medicine patients. The TAS had moderate internal reliability. In multiple regression analysis, the TAS was related to age, education, depressive symptoms, emotion suppression, self-consciousness, illness worry, and tendency to attribute somatic symptoms to psychological causes. Factor analysis of the TAS yielded four factors: Factor 1 (difficulty identifying feelings and bodily sensations) was related to education, social desirability, depressive symptoms, and private body-consciousness. Factor 2 (externally oriented thinking) was related to emotion suppression and self consciousness. Factor 3 (difficulty expressing feelings to others) was related to age, social desirability, severity of past medical illness, depressive and somatic symptoms, and emotion suppression. Factor 4 (reduced daydreaming) was related to age and self-consciousness. The TAS measures conceptually distinct dimensions that are best studied as separate factors in psychosomatic models. PMID- 8426891 TI - Coping styles in identifiers and nonidentifiers of a breast lump as a problem. AB - One hundred patients referred to a teaching hospital breast clinic for as yet undiagnosed breast masses were interviewed during their first visits and prior to assessment by a surgeon. Baseline measures of mental state and coping style were obtained. Despite the purpose of their visits, 74 of the patients were "nonidentifiers" of the breast lump as a problem on their initial contact with the clinic. All patients who were non-identifiers also used denial as a coping mechanism. Nonidentifiers used more than three times the number of avoidant coping mechanisms than "identifiers." Patients with a family history of breast cancer were more likely to be identifiers than those without a family history. PMID- 8426892 TI - Abbreviating the Duke Social Support Index for use in chronically ill elderly individuals. AB - The 35-item Duke Social Support Index (DSSI) measures multiple dimensions of social support and has been used extensively in cross-sectional and longitudinal studies of aging. Epidemiological studies of chronically ill, frail elderly individuals often wish to include a measure of social support. However, most multidimensional measures (including the DSSI) are long and may exhaust the patient, especially when included in an often already congested interview schedule. The authors have developed two abbreviated versions of the DSSI (23 item and 11-item) that capture the essential components of social support related to mental health outcomes and use of health services in treating elderly individuals with nonpsychiatric medical illness. PMID- 8426893 TI - Evaluation of pseudoseizures. A psychiatric perspective. AB - Two subgroups of patients with psychogenic pseudoseizures were identified among 20 neurology patients referred for consultation to rule out pseudoseizures. Patients with pseudo complex partial seizures were found to have primarily dissociative symptoms with guilt-laden bereavement as an important precursor. Patients with pseudo grand mal type seizures appeared to have developed their illness in the context of longstanding personality traits. Diagnostic and prognostic implications of these findings are discussed. PMID- 8426894 TI - Psychological factors affecting physical condition and DSM-IV. PMID- 8426895 TI - Potential mechanism of desipramine-related sudden death in children. PMID- 8426896 TI - Psychiatric reactions to leeches. PMID- 8426897 TI - Somnambulism associated with hallucinations. PMID- 8426898 TI - Three-dimensional CT validation of physical complaints in "psychogenic pain" patients. PMID- 8426899 TI - Coexistent hypomania and severe hypothyroidism. PMID- 8426900 TI - Study shows inconsistency in film processing quality. AB - Improper x-ray film processing is a problem for both providers and consumers of diagnostic radiology services. This article reports the findings of a study that monitored film processing practices at 297 facilities. The study showed film processing quality is inconsistent and that definitive standards need to be established for the industry. PMID- 8426901 TI - Re-entry: an untapped source of technologists. AB - Re-educating inactive radiologic technologists to re-enter practice may be an untapped source of personnel to ease the current shortage. Inactive technologists in Indiana were surveyed to learn if they were interested in returning to the work force. Results indicate that nearly 60% would consider returning to work and more than 90 percent would consider taking reentry courses. PMID- 8426902 TI - A portrait of the radiographer of the future. AB - This article describes the collection of data used by a community college in developing a plan to identify and meet the educational needs of radiographers for the year 2000 and beyond. Radiography program directors in the Midwest were asked to rank seven assumptions regarding the radiographer of the future. Respondents assigned a numerical ranking to each assumption, indicated the extent of agreement or disagreement and rated the importance of that assumption to teaching, the program and the institution. PMID- 8426903 TI - To win the battle, know your enemy. PMID- 8426904 TI - How many questions make a good exam? PMID- 8426905 TI - Evolution of glycolysis. PMID- 8426906 TI - Levels of angiotensin and molecular biology of the tissue renin angiotensin systems. AB - The cloning of renin, angiotensinogen and angiotensin converting enzyme genes have established a widespread presence of these components of the renin angiotensin system in multiple tissues. New sites of gene expression and peptide products in different tissues has provided strong evidence for the production of angiotensin independently of the endocrine blood borne system. In addition, the cloning of the angiotensin receptor (AT1) gene has confirmed the widespread distribution of angiotensin and suggested new functions for the peptide. This review of various tissues shows the variation in gene expression between tissues and angiotensin levels, and the fragmentary state of our knowledge in this area. As yet we cannot state that the gene expression of the substrates, enzymes and peptide products are involved in a single cell synthesis. This is not so much evidence against a paracrine function for tissue angiotensin, as lack of detailed, accurate intracellular information. The low abundance of renin in brain, spleen, lung and thymus compared to kidney, adrenal, heart, testes, and submandibular gland may suggest that there are both tissue renin-angiotensin systems (RAS) and nonrenin-angiotensin systems (NRAS). The NRAS could function through cleavage of angiotensinogen by serine proteinases such as tonin and cathepsin G to form Ang II directly. Although much angiotensinogen is extracellular and could therefore be a site of synthesis outside of the cell, intracellular angiotensinogen in a NRAS process could produce Ang II intracellularly without requiring extracellular conversion of Ang I to Ang II by ACE. In summary, renin mRNA is found in high concentrations in kidney, adrenal and testes and decreasing lower concentrations in ovary, liver, brain, spleen, lung and thymus. Angiotensinogen mRNA is found in the following tissues in descending order of abundance: liver, fat cells, brain (glial cells), kidney, ovary, adrenal gland, heart, lung, large intestine and stomach. It is debatable whether angiotensinogen and renin mRNA are expressed in blood vessels. The evidence that is lacking for a paracrine function of angiotensin is a complete description of the intracellular molecular synthesis and release of Ang II from single cells of promising tissues. Such tissues, SMG, ovary, testes, adrenal, pituitary and brain (neurons and glia) are potent sources of RAS components for future studies. Although the evidence for a paracrine function of angiotensin II is incomplete, it is an important concept for progressing toward the understanding of tissue peptide physiology and the significance of their gene regulation. PMID- 8426907 TI - Peripheral cholecystokinin type A receptors mediate oxytocin secretion in vivo. AB - Cholecystokinin-octapeptide (CCK8) administered intraperitoneally (i.p.) in rats induces a rapid elevation in serum oxytocin (OT). The receptor subtype mediating this action of CCK was investigated with selective CCK-A and CCK-B receptor agonists and antagonists. CCK8 and A-71623, a potent CCK-A selective agonist, were similar in efficacy and potency for stimulating OT secretion. Both compounds at 10 nmol/kg elicited approximately one-half the response of 100 nmol/kg, which elevated serum OT to approx. 20 to 30-fold above basal level. The potent CCK-B selective agonist, A-63387, at doses up to 100 nmol/kg did not increase serum OT. MK-329, a CCK-A receptor selective antagonist, at a dose of 20 nmol/kg fully inhibited the action of 20 nmol/kg CCK8, while 100 nmol/kg of (R)L-365,260, a CCK B selective antagonist, had no effect on the CCK8 response. These results, together with previous lesion studies, suggest that vagal CCK-A receptors in the periphery mediate the activation of the oxytocinergic pathway in vivo. PMID- 8426908 TI - Transforming growth factor-alpha (TGF alpha) inhibition of parietal cell secretion: structural requirements for activity. AB - Parietal cells of the gastric fundus produce transforming growth factor-alpha (TGF alpha) which functions as a potent inhibitor of acid secretion. We have previously demonstrated that TGF alpha can inhibit aminopyrine uptake in isolated rabbit parietal cells. In this study, we have evaluated the components of TGF alpha structure which determine its ability to inhibit parietal cell function. Both human and rat TGF alpha inhibited histamine stimulation by increasing the EC50 for agonist stimulation. Three fragments containing the third loop domain of TGF alpha (rat TGF alpha 34-43, human TGF alpha 34-43 and human TGF alpha 34-50) all inhibited histamine stimulation with IC50 values 20, 33 and 4-fold higher, respectively, than that of the native molecule. Rat TGF alpha inhibited carbachol stimulation throughout an agonist dose response. Human TGF alpha was only effective in inhibiting carbachol if incubations were performed in the presence of air rather than 100% O2. In air incubation, all three of the TGF alpha fragments inhibited carbachol stimulation but, in contrast to the effects on histamine, the peptides all were virtually equipotent with the native molecule. The human sequence fragments, like the native human TGF alpha, elicited no inhibition when incubations were performed in the presence of 100% O2. The results suggest that there are pharmacological differences in the response of isolated parietal cells to TGF alpha-mediated inhibition of histamine and carbachol. In addition, in contrast with previous investigations on the mitogenic action of TGF alpha, third loop fragments of TGF alpha retain the capacity to inhibit aminopyrine accumulation. PMID- 8426909 TI - Dopaminergic actions of parathyroid hormone in the rat medial basal hypothalamus in vitro. AB - Parathyroid hormone (PTH) modulates dopamine (DA) metabolism in the rat medial basal hypothalamus (MBH) in vivo. Direct effects of PTH on MBH DA metabolism were therefore investigated in vitro. Incubation of rat MBHs for 60 min with 10(-7) 10(-5) M human PTH1-34 consistently reduced the tissue DA content and increased the DOPAC (dihydroxyphenylacetic acid) to DA ratio. This ratio was further increased in tissues incubated in 10(-5) M PTH1-34, as a result of an increase in DOPAC content. The tissue content of DOPAC and DA was unaffected by 10(-9) M PTH. The serotonin (5HT) content of the MBH was reduced by 10(-5) M PTH1-34, but concentrations of 5HT, 5-hydroxyindolacetic acid, and norepinephrine were otherwise unaffected by 10(-9)-10(-5) M PTH1-34. Concentrations of DA in the incubation media were reduced after exposure to 10(-6) or 10(-5) M PTH1-34. The uptake of 3H-labelled DA by incubated tissues was also reduced by 10(-6) M PTH1 34, as was the metabolism of 3H-labelled DA into tissue and media DOPAC. Monoamine oxidase (MAO) activities A and B were significantly increased after the incubation of the MBH with 10(-6) or 10(-5) M PTH1-34. These results further demonstrate neuromodulatory actions of PTH on dopaminergic neurons within the rat MBH in vitro, and suggest neural and/or neuroendocrine roles of PTH of central or peripheral origin. PMID- 8426910 TI - Identification of IGF-1 receptors in primitive vertebrates. AB - This is the first report of the existence of insulin-like growth factor (IGF-1) receptors in three representatives of lower vertebrates: the osteichtyes, chondrichtyes and cyclostomi. Competitive binding studies and affinity labelling of brain membranes from Cottus scorpius (sea scorpion), Raja clavata (ray) and Myxine glutinosa (atlantic hagfish) identified a mammalian type 1 or IGF-1 receptor by its binding specificity and the molecular size of its alpha-subunit. IGF-1 and IGF-2 are almost equally potent in displacing receptor-bound 125I-IGF-1 or 125I-IGF-2, and the proteins labeled with both tracers have a molecular size of 100,000-120,000 under reducing conditions. There was no evidence for the presence of a mammalian type 2 or IGF-2/mannose 6-phosphate receptor in brains of Cottus, Raja or Myxine. In all three species the binding of 125I-IGF-1 and 125I IGF-2 was significantly higher in brain compared with liver and gastrointestinal tract, and the IGF-1 receptor could only be identified with certainty in Raja liver. It is concluded that the brain of three lower vertebrates express mammalian IGF-1 receptors, whereas IGF-2-mannose 6-phosphate receptors could not be detected. PMID- 8426911 TI - Effects of a physiological dose of cholecystokinin on food intake and postprandial satiation in man. AB - CCK-33 was infused intravenously to groups of 9 lean and 9 obese volunteers in doses that elicited plasma CCK concentrations in the physiological range. The effect of these infusions on food intake and satiety signals was compared with the effect of saline infusions in the same subjects. Food intake (486 +/- 52 g; mean +/- S.E.M.) was slightly, but not significantly decreased (553 +/- 55 g after saline), and hunger and fullness feelings after eating were unaffected, in both of the two groups. We conclude that the infusion of CCK-33 to plasma levels comparable to those observed after a fatty meal does not have a major effect on food intake and postprandial hunger feelings. PMID- 8426912 TI - Amylin inhibits glucose-induced insulin secretion in a dose-dependent manner. Study in the perfused rat pancreas. AB - Islet amyloid polypeptide (IAPP), also called amylin, has been localized in the B cell secretory granule and is co-secreted with insulin. We have investigated the effect of synthetic amidated rat amylin on the insulin release evoked by 9 mM glucose in the isolated, perfused rat pancreas. Amylin, in a range of 75 nM-75 pM, significantly inhibited this insulin response in a dose-dependent manner. The correlation between the logarithm of amylin concentrations and the percentages of inhibition was highly significant (r = 0.98, P < 0.01). The lowest effective amylin concentration tested (75 pM) is within the range of amylin levels reported for the effluent of the perfused rat pancreas. Finally, pre-infusion of the rat pancreas with a high amylin concentration (75 nM) did not alter the insulin response to glucose, thus excluding a toxic effect of amylin on the B-cell. These observations support the concept that amylin plays a role in the control of insulin secretion. PMID- 8426913 TI - Tissue-specific processing of neurotensin/neuromedin-N precursor in cat. AB - Antisera were raised towards the putative N-terminal sequence of the canine precursor to neurotensin (NT) and neuromedin N (NMN), residues 24-35 in the pre prohormone, as well as its C-terminal TAIL, residues 164-170. These were used in conjunction with previously developed immunoassays towards NT and NMN to characterize the precursor and to study its processing in feline brain, heart, adrenal and intestine. The precursor was identified as a 18 kDa protein that reacted with both the N- and C-terminal antisera and yielded NT3-13 and NT4-13 upon treatment with pepsin. It represented 5-10% of the NT activity in ileum. Processing was found to be tissue-specific, yielding primarily NT, NMN and TAIL in most tissues but also giving rise to N-terminally extended forms of these peptides, which were particularly evident in extracts of adrenal and intestine. Two of the four molecular forms detected using the TAIL immunoassay were tenatively identified as NT-TAIL and Gly-Ser-Tyr-Tyr-Tyr based upon chromatographic evidence. Western blotting indicated that ileal extracts contained large molecular forms of NT (17 kDa) and NMN (16 kDa), whereas brain extracts contained primarily the N-terminal of the precursor without NT and NMN attached (14.3 kDa). It was concluded that the structure of the feline precursor in its N- and C-terminal regions, and the positioning of the signal peptide cleavage site were as predicted from cDNA work in the dog. On the other hand, processing of the precursor was complex, occurring in a tissue-specific manner. PMID- 8426914 TI - Complications of laparoscopic cholecystectomy and their interventional radiologic management. AB - Cholelithiasis and cholecystitis are encountered throughout the world and are responsible for the majority of cholecystectomies performed. Treatment has traditionally consisted of open cholecystectomy, but laparoscopic cholecystectomy is currently a popular alternative. Laparoscopic cholecystectomy offers several benefits over the open procedure, but it also has its own set of complications. Complications include those of laparoscopy (abdominal wall bleeding, omental bleeding, abdominal vessel injury, retroperitoneal vessel injury, gastrointestinal perforation, bladder perforation, solid visceral injury, and infection) and those of cholecystectomy (gallbladder fossa bleeding, bile duct injury, bile leakage, and infection). The literature suggests that the total complication rate for the laparoscopic procedure compares favorably with that of the open procedure, but this may apply only to surgeons who have accomplished numerous laparoscopic procedures and not to those who have just completed a training course and are performing their first few procedures. With the growing success of the laparoscopic procedure, it is essential that the radiologist be knowledgeable about the radiographic manifestations and interventional radiologic management of potential complications, since the radiologist can significantly affect patient care. PMID- 8426915 TI - Fundamental properties of digital images. AB - The quality of a digital image is affected by matrix size, unsharpness in the underlying image, bit depth, and noise in the underlying image. The array of all of the pixels into which an image is divided is the image matrix. If the image matrix is small (ie, composed of a few large pixels), the resolution of the digital image is low. Image blur dominates when the pixel size is much smaller than the unsharpness of the underlying image. The density in a pixel is represented by a binary number with a variable number of bits (bit depth). The more noise present in the initial image, the fewer the bits that are needed to convey its representation. Insufficient numbers of bits in the digital representation of an image, whether from too few pixels or too small a bit depth in each pixel, result in inadequate image quality. Use of too many bits, relative to the quality of the underlying image, places an increasingly unwarranted demand on digital image handling and processing. PMID- 8426916 TI - Historical perspective on computer development and glossary of terms. AB - This article contains a concise history of the development of mechanical and electronic computers, descriptions of the milestones in software development, discussion of the introduction and adoption of computers in radiology, and a glossary of computer terms used frequently in radiology. One of the earliest devices designed to mechanize calculations was the calculating clock, built in 1623. The first programmable electronic computer, the ENIAC (electronic numerical integration and computer), was completed in 1945 at the University of Pennsylvania. Software has developed from early machine language through fourth generation languages and graphic user interfaces used today. The computer was introduced to radiology initially in the 1960s in nuclear medicine and is now incorporated in many digital imaging modalities throughout radiology. The development of picture archiving and communication systems has resulted in the implementation of several totally digital departments of radiology. PMID- 8426917 TI - The computer in the radiologist's office. AB - Today, the radiologist is able to equip his or her office with a powerful personal computer system equivalent to the large mainframe computers of just a few years ago. If funds permit, purchase of a 486 equivalent system with 200-300 Mbytes of hard-disk storage, 16 Mbytes of random access memory (RAM), a high resolution color video card and monitor, and a laser printer is recommended. The practical uses for such a system are almost limitless and include word processing, spreadsheet and data-base management, telecommunications, multimedia presentations, business applications, teleradiology, resident and medical student education, and research applications. No matter how much one becomes involved in computer applications, it is essential to establish good habits for backing up critical data files and programs. Becoming familiar with computer technology is not easy at first. Finding a good computer buddy, taking simple night school courses, and reading computer articles and magazines are good ways to get started. Computers are wonderful devices. The day is fast approaching when they will become a necessary tool for every radiologist. PMID- 8426918 TI - Advanced applications of personal computers in the radiologist's office. AB - The author's department has found various advanced applications for the computer to be useful in daily practice. They use a data-base program to track interesting cases for later retrieval. The program automatically generates an American College of Radiology code based on the body part and diagnosis. The program is also used to track radiographic film quality. A barcode scanner attached to a computer at the film alternator is used to enter the accession number generated by the radiology information system. If any deficiencies are present, they are entered from a preprinted bar-code sheet. The bar-code scanner allows rapid entry of all examinations during the read-out session. Reports generated from the data base have been helpful in identifying and quantifying radiographic examination deficiencies. Department computers are also connected to the campus Ethernet network. This network allows radiologists to electronically verify radiology reports and to conduct electronic literature searches on the computers in their offices. PMID- 8426919 TI - Image interpretation session: 1992. Esophageal strictures and squamous cell carcinoma of the maxillary sinus and palate in recessive epidermolysis bullosa dystrophica. PMID- 8426920 TI - Image interpretation session: 1992. Pott disease (tuberculous spondylitis), with alcoholic cirrhosis and an incidental fibroadenoma of the left breast. PMID- 8426921 TI - Image interpretation session: 1992. Tracheopathia osteoplastica. PMID- 8426922 TI - Image interpretation session: 1992. von Hippel-Lindau disease. PMID- 8426923 TI - Image interpretation session: 1992. Recurrent adenocarcinoma of the esophagus with direct invasion into the pulmonary veins and left atrium resulting in both septic and air embolism (Fig 20). Tumor also invaded the right lower lung, resulting in postobstructive pneumonia. PMID- 8426924 TI - Image interpretation session: 1992. Ewing sarcoma with metastases to the lung. PMID- 8426925 TI - Image interpretation session: 1992. Torsion of the right upper lobe. PMID- 8426926 TI - Image interpretation session: 1992 Tuberculoma of the left frontoparietal lobe, with relapse after cessation of treatment. PMID- 8426927 TI - Image interpretation session: 1992. Gastric duplication cyst (lined with gastric mucosa) with a tract (lined with pancreatic tissue) leading to the pancreas (Fig 38). PMID- 8426928 TI - Ampullary tumors: radiologic-pathologic correlation. AB - Ampullary adenoma and adenocarcinoma are the benign and malignant neoplasms that arise from the glandular epithelium of the ampulla of Vater. When the tumors are small and confined to the ampulla, endoscopic and radiographic appearances may be normal. Larger intraampullary tumors appear as a prominent papilla or a mural mass. Computed tomography and ultrasound (US) show dilatation of the common bile duct or pancreatic duct in such cases, but the mass itself may not be seen. Larger lesions with intraduodenal extension are more easily identified with cross sectional imaging. Endoscopic US is the best technique for tumor staging. The differential diagnosis includes other periampullary tumors such as pancreatic carcinoma, cholangiocarcinoma, and villous adenomas and adenocarcinomas of the duodenum. Mural masses of the periampullary duodenum including choledochocele and carcinoid should also be considered. Accurate diagnosis of ampullary tumors is important because the patients are usually candidates for a Whipple operation. Recent reports quote overall 5-year survival rates of 28%-70%. The prognosis is even better for patients with a limited stage of disease. PMID- 8426929 TI - MR imaging of the developing human brain. Part 1. Prenatal development. AB - To establish a baseline of the magnetic resonance (MR) imaging appearance of the fetal brain in early stages of development, the authors undertook a study of fixed and fresh specimens of embryos and fetuses of 6-28 weeks gestational age. Images of formalin-preserved and fresh specimens were comparable in their depiction of anatomic structures. On MR images of embryos of 6 weeks gestational age, the rhombic and cervical flexures, aqueduct of Sylvius, diencephalon, cerebellum, cerebral hemisphere, and fourth ventricle could be differentiated. The optic recess and chiasm, pituitary gland, pineal recess, third ventricle, pons, olfactory lobe, corpus striatum, insula, and parietal and temporal lobes could be distinguished as early as 11 weeks gestation. Although MR imaging is impractical as a screening tool for intrauterine abnormalities, it can demonstrate the fetus in great detail and allows a more specific evaluation of fetal anatomy. With the information provided by MR imaging, it may be possible to establish guidelines for assessment of the stage of development during intrauterine life. PMID- 8426930 TI - Ultrasound case of the day. Gallbladder varices. PMID- 8426931 TI - Pediatric case of the day. Duplication cyst of the distal ileum, acting as a lead point for intussusception. PMID- 8426932 TI - General case of the day. Acute lymphoblastic leukemia. PMID- 8426933 TI - Image quality in dynamic CT: a clinical discussion. AB - Modern state-of-the-art computed tomographic (CT) scanners emphasize three capabilities: image quality, dynamic scan capability, and a high-resolution thin section technique. Image quality is fundamental and dependent on optimum performance and the interrelationship of all system components. Variables that affect the performance of the scanner include x-ray tube output and rate of heat dissipation; quantum detection efficiency; electronic noise in the acquisition system; speed, accuracy, and integration of mechanical motion in the gantry and table; and the algorithm used for image reconstruction. System design must allow for dynamic scan operation, either in the single-scan or cluster mode, with short interscan or intergroup delays or, as more recently developed, with helical acquisition. Dynamic scanning is frequently used for nonneurologic applications, including diagnosis of vascular and perivascular diseases and multifocal organ disease, particularly hepatic disease. Efficient operation depends on rapid reconstruction and display capability. Modern systems have been engineered to provide flexible modes of operation, particularly in dynamic scanning, and rapid on-line review and analysis, all of which serve to improve the quality of images produced with dynamic CT scanning. PMID- 8426934 TI - The scientific exhibit in radiology. PMID- 8426935 TI - MR imaging of the hands in early rheumatoid arthritis: preliminary results. AB - Several magnetic resonance (MR) imaging pulse sequences (unenhanced T1 weighted spin echo, with and without fat suppression; unenhanced T2 weighted with fat suppression; and spoiled gradient-recalled acquisition in the steady state [GRASS], with fat suppression, before and after gadolinium enhancement) were analyzed in 30 patients with early rheumatoid arthritis to determine which sequences were best for imaging various aspects of the disease as manifested in the hands and wrists. A single hand was imaged in all patients. Inflammation and erosions were best seen on spoiled GRASS images with fat suppression and gadolinium enhancement, with the latter also being well seen on T1-weighted images. Articular cartilage was best seen on unenhanced spoiled GRASS or T1 weighted images with fat suppression. For the latter application, increased resolution is needed before the results will be meaningful. MR imaging has potential as an objective method for evaluating rheumatoid arthritis. However, change in MR imaging findings in a single hand must be carefully monitored and compared with the overall disease course for a long period in a large number of patients to establish the clinical value of MR imaging. PMID- 8426936 TI - Abnormal air-filled spaces in the lung. AB - The authors reviewed the radiographic and computed tomographic (CT) appearances of abnormal air-filled spaces in the lung that develop in response to lung diseases. The major types of these lung diseases include infection, vessel related or vascular-embolic disorders, bronchiectasis, emphysema, pulmonary fibrosis, adult respiratory distress syndrome and air-block diseases, and unusual disorders of the lung (such as Langerhans cell histiocytosis, Klippel-Trenaunay syndrome, and tracheolaryngeal papillomatosis). After studying the CT scans, conventional radiographs, and medical records of 150 patients with various abnormal air-filled spaces in their lungs and 300 lung specimens and the corresponding high-resolution CT scans, the authors concluded that mechanisms of air-space formation fall into five basic categories: (a) vascular occlusion or ischemic necrosis, (b) dilatation of the bronchi, (c) disruption of the elastic fiber network of the lung, (d) remodeling of the lung architecture and retractile fibrosis, and (e) multifactorial or unknown mechanisms. PMID- 8426937 TI - Comparison of time-of-flight and phase-contrast MR neuroangiographic techniques. AB - Magnetic resonance (MR) angiography constitutes a group of evolving MR imaging techniques that can be used to directly image flow in arteries, veins, and cerebrospinal fluid. By using multiple MR angiographic sequences in each of 18 patients with neurologic conditions such as aneurysms, arteriovenous malformations, and carotid atherostenotic disease, the advantages, disadvantages, and unique artifacts of each technique could be investigated. Time-of-flight imaging is susceptible to saturation effects, and short T1 substances may simulate flow. Two-dimensional time-of-flight imaging is useful in cranial venography in assessing the patency of the dural sinuses or venous drainage from an arteriovenous malformation. Three-dimensional time-of-flight images depict small and medium-sized aneurysms. Phase-contrast imaging has excellent background suppression, allows variable velocity encoding, and provides directional flow information. Two-dimensional phase-contrast imaging is useful in the assessment of the patency of major vascular structures. Three-dimensional phase-contrast imaging (with 30-cm/sec velocity encoding) is also useful in depicting small and medium-sized aneurysms. Although some applications of MR angiography are clear, further study and development of the techniques are necessary to determine which imaging sequence or combination of techniques is best suited for answering specific diagnostic questions. PMID- 8426938 TI - Asymmetric radiographic findings in the pediatric chest: approach to early diagnosis. AB - Several conditions in children produce a mass effect within one hemithorax, which may be life threatening. The different radiographic appearances of these conditions provide a basis on which they can be classified. The three major categories include a large cystic hemithorax, a large lucent hemithorax, and a large opaque hemithorax. Some disorders may be seen within more than one major group. Within the large cystic hemithorax group, congenital cystic adenomatoid malformation and congenital diaphragmatic hernia may be life-threatening conditions that require emergency surgery. The large lucent hemithorax is usually caused by pneumothorax, partial bronchial occlusion, or compensatory hyperinflation of the unobstructed lung. The majority of cases within the large opaque hemithorax group involve pleural fluid collections. Because many conditions that cause a mass effect require intervention or surgery, accurate interpretation of the plain chest radiograph is essential for an early diagnosis. Classification of radiographic features into these three groups often allows accurate diagnosis without additional studies. PMID- 8426939 TI - Breast implants, common complications, and concurrent breast disease. AB - Recent concern regarding breast implants has emphasized the special imaging needs of the approximately 2 million American women who currently have prosthetic breast implants, including silicone gel, silicone gel with a textured silicone coating, saline, biocompatible gel, polyurethane-coated, double-lumen, and tissue expanders. Both palpable and nonpalpable breast lesions can occur in patients with implants, and these lesions must be evaluated in the same manner as in patients without implants, which presents a challenge for the mammographer. Not only is the breast tissue of a patient with implants more difficult to image, but the patient may have complications from the implants. The major complications of their use involve hematoma in the early postoperative period, infection, capsule contracture, rupture, and silicone granulomas. Familiarity with the more common types of implants, the possible complications of their use, and concurrent breast disease may help improve diagnosis in these patients. PMID- 8426940 TI - Epilepsy, sports and exercise. PMID- 8426941 TI - Caffeine and exercise performance. An update. AB - Three principal cellular mechanisms have been proposed to explain the ergogenic potential of caffeine during exercise: (a) increased myofilament affinity for calcium and/or increased release of calcium from the sarcoplasmic reticulum in skeletal muscle; (b) cellular actions caused by accumulation of cyclic-3',5' adenosine monophosphate (cAMP) in various tissues including skeletal muscle and adipocytes; and (c) cellular actions mediated by competitive inhibition of adenosine receptors in the central nervous system and somatic cells. The relative importance of each of the above mechanisms in explaining in vivo physiological effects of caffeine during exercise continues to be debated. However, growing evidence suggests that inhibition of adenosine receptors is one of the most important, if not the most important, mechanism to explain the physiological effects of caffeine at nontoxic plasma concentrations. Numerous animal studies using high caffeine doses have reported increased force development in isolated skeletal muscle in both in vitro and in situ preparations. In contrast, in vivo human studies have not consistently shown caffeine to enhance muscular performance during high intensity, short term exercise. Further, recent evidence supports previous work that shows caffeine does not improve performance during short term incremental exercise. Although controversy exists, the major part of published evidence evaluating performance supports the notion that caffeine is ergogenic during prolonged (> 30 min), moderate intensity (approximately 75 to 80% VO2max) exercise. The mechanism to explain these findings may be linked to a caffeine-mediated glycogen sparing effect secondary to an increased rate of lipolysis. PMID- 8426942 TI - Physiological and biomechanical aspects of rowing. Implications for training. AB - The drag force on a racing shell increases with the square of velocity corresponding to a 3.2 power increase in energy expenditure. However, the metabolic cost increases with only an approximately 2.4 power function of shell velocity. During international races the metabolic cost corresponds to an oxygen uptake of 6.7 to 7.0 L/min over 6.5 min. The relative anaerobic contribution to 6.5 min of 'all-out' rowing has not been determined but is estimated to range from 21 to 30%. Because of the large muscle mass involved in rowing, blood variables reach extreme values: adrenaline 19 nmol/L; noradrenaline 74 nmol/L; pH 7.1; and bicarbonate 9.8 mmol/L. Because of the static component of the rowing stroke at the catch, blood pressure increases to near 200mm Hg, and the heart of oarsmen has adapted to this load by increasing wall thickness and internal diameters. The maximal oxygen uptake of oarsmen may reach 6.6 L/min and ventilation 243 L/min. Arterial oxygen tension decreases by 20mm Hg during 'all out' rowing corresponding to a decrease in pulmonary diffusion capacity. A force of approximately 800 to 900N is developed on the oar. Force generation during rowing is relatively slow, 0.3 to 0.4 sec. Oarsmen are strongest in low velocity movement with 70 to 75% slow twitch fibres in skeletal muscle. Data indicate that rowing technique and training may improve explaining why results become approximately 0.7 sec faster per year. PMID- 8426944 TI - Human immunodeficiency virus infection, exercise and athletics. PMID- 8426943 TI - Dietary supplements in sport. AB - Studies of the dietary practices of athletes report that nutritional supplements are commonly used. Supplementation practices vary between sports and individual athletes; however, there is evidence that at least some athletes use a large number of supplements concurrently, often in doses that are very high in comparison with normal dietary intakes. In exploring supplementation practices we propose a classification system separating the supplements into dietary supplements and nutritional erogogenic aids. The dietary supplement is characterised as a product which can be used to address physiological or nutritional issues arising in sport. It may provide a convenient or practical means of consuming special nutrient requirements for exercise, or it may be used to prevent/reverse nutritional deficiencies that commonly occur among athletes. The basis of the dietary supplement is an understanding of nutritional requirements and physiological effects of exercise. When the supplement is used to successfully meet a physiological/nutritional goal arising in sport it may be demonstrated to improve sports performance. While there is some interest in refining the composition or formulation of some dietary supplements, the real interest belongs to the use or application of the supplement; i.e. educating athletes to understand and achieve their nutritional needs in a specific sports situation. The sports drink (carbohydrate-electrolyte replacement drink) is a well known example of a dietary supplement. Scientific attitudes towards the sports drink have changed over the past 20 years. Initial caution that carbohydrate-electrolyte fluids compromise gastric emptying during exercise has now been shown to be unjustified. Numerous studies have shown that 5 to 10% solutions of glucose, glucose polymers (maltodextrins) and other simple sugars all have suitable gastric emptying characteristics for the delivery of fluid and moderate amounts of carbohydrate substrate. The optimal concentration of electrolytes, particularly sodium, remains unknown. Most currently available sports drinks provide a low level of sodium (10 to 25 mmol/L) in recognition that sodium intake may promote intestinal absorption of fluid as well as assist in rehydration. The sodium level of commercial oral rehydration fluids (used in the clinical treatment of diarrhoea and dehydration) is higher than that of the present range of sports drinks. However, even if research indicates that intestinal glucose transport is optimally stimulated at higher sodium concentrations, concern for the palatability of sports drinks may impose a lower ceiling for sodium levels. Commercial viability of a sports drink requires that it provide a refreshing and palatable fluid replacement across a wide variety of sports and exercise situations.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8426945 TI - Treatment of lower extremity injuries with orthotic shoe inserts. An overview. AB - Orthotic shoe inserts are very effective in providing symptomatic relief of lower extremity complaints in running athletes. Inserts adjust the biomechanical variables associated with running injuries and reduce the effect of high stresses produced by running activities. Orthotic treatment is based on an understanding of complex coupling of rotation of the lower extremity with pronation and supination of the subtalar joint. Orthotic fabrication is initiated by determining the neutral position of the foot and obtaining an accurate cast of this position. Successful treatment with orthotic shoe inserts is dependent on careful evaluation of the runner and formulation of a properly fitted orthosis. When correctly utilised, orthotic shoe inserts are beneficial for a broad range of disorders experienced by runners. Since biomechanical deficits may be related to injuries along the entire lower extremity, specific diagnoses may be of lesser indication; however, accurate identification of the underlying biomechanical deficit is critical. Problems related to excessive or prolonged pronation are most amenable to orthotic treatment. While treatment of the cavus foot with orthotic shoe inserts is sometimes worthwhile, the clinician should be aware of limited success in this instance. Finally, orthotics are only one facet in the overall treatment plan for injured running athletes. Most overuse syndromes will respond to rest, training modification, and a change in the running surface or shoe. Equally important is the use of a proper conditioning and stretching programme both for injury prevention and for treatment of specific injuries. Treatment with orthotic shoe inserts should not be used as a substitute for any of these approaches. PMID- 8426946 TI - [Erythrocyte CR1 receptor: binding and transport of immune complexes in the blood circulation]. AB - Immune complexes (IC), after reacting with the complement system, bear C3b fragments (opsonized IC) and bind to the CR1 receptor that is present on human erythrocytes (CR1 = complement receptor type 1). This efficient binding reaction prevents random vascular IC deposition, and allows IC to be transported through the circulation to the fixed macrophage system of the liver and spleen, where they are safely eliminated. The structure of CR1, with multiple C3b binding sites, and the clustered distribution of CR1 on the erythrocyte surface favor the multivalent binding of opsonized IC to erythrocytes. CR1 on erythrocytes serves as a cofactor for the inactivation of C3b by factor I, thus allowing the release of IC from the erythrocyte surface and their transfer to fixed macrophages. Under normal circumstances, the erythrocyte plays a major role in the processing of IC in humans. PMID- 8426947 TI - [The role of cytokines in normal and in leukemic hematopoiesis]. AB - There have been astonishing advances in the field of cytokines in recent years. In the present paper we focus on a few hematopoietic cytokines and discuss some aspects of the biology and pathophysiology of cytokines and their receptors in normal and leukemic hematopoiesis. We also address future developments and in particular the role of negative stem cell regulators. PMID- 8426948 TI - [Polycythemia: primary or secondary? The differential diagnostic value of stem cell cultures]. AB - Cultures of hematopoietic precursor cells can be helpful in differentiating between primary polycythemia (polycythaemia vera, PV) and reactive secondary polycythemia: in PV erythroid precursors form hemoglobinized colonies in the absence of added erythropoietin (epo) (= endogenous erythroid colonies), whereas in normals and in patients with secondary polycythemia, formation of erythroid colonies is dependent on added epo. We have performed cultures of peripheral blood precursors from 132 patients with elevated hemoglobin in the presence/absence of added epo. In 48/132 patients we assumed that PV was the cause of polycythemia. In 80/132 patients no endogenous colonies appeared and the polycythemia was judged secondary. 23 PV patients were examined repeatedly. In 18 of them the first diagnosis was confirmed by subsequent cultures; in 5 cases endogenous colonies, which had been present in the first cultures, were no longer detectable. A questionnaire on the subsequent clinical course was sent to 108 treating physicians. 77 questionnaires were answered correctly and returned. In 86% of these patients, our culture diagnosis of PV was either confirmed or another myeloproliferative disorder had been found as a cause of endogenous colonies. In 14% our diagnosis of PV had been false positive. On the other hand, our diagnosis of secondary polycythemia was confirmed in 85% of the patients; its most frequent cause was cigarette smoking and chronic bronchitis and only rarely was it associated with heart or kidney disease. 5/77 patients had persistently elevated Hb without an evident cause, and in 3/77 the Hb normalized spontaneously.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8426949 TI - [Autoimmune hemolytic anemia with medullary erythroplasia and black serum with methemalbuminemia]. AB - A patient with idiopathic autoimmune hemolytic anemia is described. On day 5 of therapy with corticosteroids, immunoglobulin and several plasmaphereses, a high level of methemalbumin was found while RBC lysis had ceased and bilirubin and haptoglobin had normalized. The hemoglobin and the reticulocyte count were still very low and a bone marrow aspirate showed erythroaplasia. Since the LDH was consistently elevated during the entire period of erythroaplasia, ongoing intramedullary hemolysis due to antibodies (IgG) was assumed. The patient received cyclophosphamide. After 15 days all symptoms, signs of hemolysis and aplasia resolved. The patient has not had any other hemolytic crisis without treatment (follow-up 18 months). The presence of methemalbumin is observed in intravascular hemolysis and also in association with hemorrhagic pancreatitis. Its diagnostic and prognostic value is discussed. PMID- 8426950 TI - [Initial experiences with a recombinant human tissue thromboplastin in oral anticoagulation]. AB - A recombinant tissue factor (rTF, Batch TFS-RD3, Baxter Diagnostics) has been assessed by plasma and capillary blood methods and compared with our working rabbit brain thromboplastin (CRB Roche, Basel, Switzerland) for its suitability in monitoring oral anticoagulant therapy. The international sensitivity index (ISI) for rTF calibrated against CRB has been found to be 0.73 and 0.69 respectively. The slope of the orthogonal regression line between log prothrombin time rTF plasma and rTF capillary blood is 0.96. Thus, we considered the ISI to be identical for both methods. International normalized ratios calculated with these assumptions, and even activity percentages assigned by a dilution curve, did not differ from the results obtained with our working thromboplastin. The human recombinant thromboplastin TFS-RD3 seems to have properties similar to currently used commercial thromboplastins. Recombinant tissue factor as a well defined preparation could make essential contributions to the standardization of prothrombin time and to the improvement of oral anticoagulant therapy. PMID- 8426951 TI - [Childbirth without fear in case of Von Willebrand's disease]. AB - We report two cases of von Willebrand (type I) disease with vWF antigen levels of 21 and 25%, vWF ristocetin cofactor concentrations of 32 and 30%, and coagulation FVIII levels of 10 and 50% respectively. Template bleeding times were 20 and 11 minutes respectively. Both of these women exhibited a progressive and marked improvement of the vWF/FVIII complex during pregnancy. In the first, all parameters had completely normalized in the 37th week when a cesarean section was performed without hemorrhagic complications. The second case had borderline normal levels of the vWF/FVIII complex in the 37th week and underwent normal delivery in the 39th week. The improvement persisted for at least one week post partum and in the first case values were still higher 10th weeks after delivery than they had been prior to the pregnancy. These observations and similar cases in the recent literature suggest that obstetrical operations and normal delivery are not associated with major blood losses in cases of moderately severe vWD of type I. These patients probably will not need vWF/FVIII concentrates for adequate hemostasis. PMID- 8426952 TI - [Factor VII and protein-C markers are no prognostic indicators in acute coronary heart disease]. AB - Fibrinogen and factor VII have been identified as independent risk factors in cardiovascular morbidity. A relationship between factor VII coagulant activity (VII:C) and prognostically different manifestations of acute coronary heart disease (aCHD) has recently been suggested. In order to validate and to extend these observations, we prospectively studied patients admitted for aCHD (n = 76) and a control group without aCHD (CG, n = 27). According to their clinical evolution, the aCHD cases were subdivided into unstable angina (UA, n = 26), myocardial infarction without (M-I, n = 23) and with heart failure (M-II, n = 27). Before treatment blood was collected for the following assays: fibrinogen, factor VII procoagulant activity (VII:C), amidolytic test (VII:am), immunological (VII:Ag), protein C functional (PCf) and immunological (PC:Ag). We found no statistically significant difference between control and aCHD cases and between their subgroups for any assay of fibrinogen, factor VII and protein C. The VII:C/VII:Ag ratio was higher for UA, M-I, M-II and the entire aCHD group compared with the CG. However, it was not possible to separate the prognostically different aCHD subgroups from each other by ratios of measured values. Therefore, determinations of factor VII, protein C and fibrinogen in aCHD have no prognostic relevance. PMID- 8426953 TI - The impact of the HLA-system in clinical medicine. AB - A short overview of the genetics of HLA is given. The most remarkable point of the present day situation is that when we use HLA in the clinic we not only ignore much of the immunogenetic information which is available, but that on top of that we are still ignorant of a large number of genes in the HLA super-region and of their functions. When reviewing the biology of HLA it becomes clear that evidence can be found which supports the notion that the polymorphism might protect the human race against eradication by epidemics or endemics. The molecular basis of the recognition of viral, bacterial or other allogeneic antigens is briefly and schematically summarized. The clinical importance of HLA s.s. is two-sided. The first and at the moment in the clinic the most important contribution is matching for HLA in platelet transfusion, bone marrow transplantation and organ transplantation. The other reflects on the capacity of allogeneic HLA antigens, whether given as blood transfusion or through exposure of the newborn infant to maternal cells to influence the T and B cell repertoire. Preliminary evidence is reviewed indicating that this might not only be of importance in organ transplantation but also in disease predisposition. PMID- 8426954 TI - [Granting of the 72d Marcel-Benoist Foundation Award]. PMID- 8426955 TI - [Inadequate treatment compliance, patient information and drug prescription as causes for emergency hospitalization of patients with chronic heart failure]. AB - Causes of decompensation of treated chronic congestive heart failure in patients referred for emergency hospitalization were examined prospectively. 111 consecutive patients (76 +/- 11 years) were interviewed and their records examined on admission. The diagnosed underlying diseases were coronary artery disease (80%), hypertensive heart disease (40%), valvular heart disease (11%), and idiopathic dilated (7%) and alcoholic (5%) cardiomyopathy. The grounds for decompensation of chronic congestive heart failure were: insufficient compliance 47% (n = 52, irregular or not intake of medication [25%], salt [9%] or fluid [7%] excess, stopping medication because of side effects [6%]), uncontrolled hypertension (27%), insufficient diuretic therapy in spite of progressive symptoms (23%), treatment with negative inotropic drugs (21%), acute rhythm disturbances (14%), acute myocardial infarction or unstable angina pectoris (14%), infections (6%). 80% of the patients were treated with diuretics, 34% with digoxin, 31% with ACE-inhibitors. Insufficient basic knowledge about the disease (regular weighing, diet, behavior if symptoms worsen) was found in 78% of patients, complete lack of knowledge concerning the prescribed drugs in 29%. Only 44% were regularly followed by their physicians, 53% had either no regular follow ups or they were set at too long intervals. CONCLUSIONS: In the majority of patients, one or more avoidable causes leading to decompensation of chronic congestive heart failure can be identified. The main potential for intervention aiming at a reduction of the hospitalization frequency lies in improving patient compliance and state of the art medication by the primary care physician. Equally unsatisfactory is the low frequency of follow-up checks to reassess and renew drug therapy. PMID- 8426956 TI - [Latex allergy--an increasing problem in clinical practice]. AB - In the last few years the allergenic potential of rubber has been receiving more and more attention. Its widespread application in medical devices and everyday objects has increased exposure to this allergen. The central role of rubber in the prevention of HIV infection has certainly played a part in this development. Latex allergy manifests itself as immediate type allergy (type I), delayed type allergy (type IV) or as a combination of both types of hypersensitivity. In this review the different types of latex allergy are illustrated by means of selected case reports. Immediate type allergy to latex leads to contact urticaria, but it may also cause generalized urticaria, bronchial asthma or even anaphylactic reactions (contact urticaria syndrome). Hence, not only dermatologists and allergologists are confronted with latex allergy. Immediate type hypersensitivity can be proven by skin prick test and RAST (radio allergosorbent test). Contact dermatitis in patients using rubber gloves is generally due to delayed type hypersensitivity to rubber additives. However, a recently observed case suggests the possibility of isolated delayed type hypersensitivity to natural latex. PMID- 8426957 TI - [Imported dengue fever following a stay in the tropics]. AB - We report on a tourist returning from Thailand, who presented with classical dengue fever. While in Thailand a 36-year-old Swiss female laboratory assistant suddenly developed fever, devastating headache, retro-ocular pain, myalgia and arthralgia, photophobia, nausea and diarrhea. In addition she suffered from epistaxis, urogenital and skin bleeding, and a morbilliform exanthema. After her return to Switzerland we noted lymphadenopathy and splenomegaly, enanthema and laboratory findings of mild hepatitis, thrombocytopenia and leukopenia. The diagnosis of dengue virus infection was verified serologically. Apart from a long lasting convalescent asthenia we observed restitutio ad integrum within days under symptomatic therapy. Epidemiological clinical and diagnostic aspects of dengue virus infection are discussed. PMID- 8426958 TI - [Grand-mal epilepsy as initial manifestation of a parenchymal neuro_cysticercosis]. AB - The case of a 29-year-old male immigrant from India with parenchymal neurocysticercosis is reported. Epileptic seizures were the first clinical manifestation of the disease, which was successfully treated with praziquantel. The follow-up covers a period of three years. PMID- 8426959 TI - [A heart attack]. PMID- 8426960 TI - Guide to scientific products, instruments, and services. Revised product listing. PMID- 8426961 TI - Why changing the U.S. health care system is so difficult. PMID- 8426962 TI - The competing discourses of HIV/AIDS in sub-Saharan Africa: discourses of rights and empowerment vs discourses of control and exclusion. AB - The competing discourses of HIV/AIDS circulating in sub-Saharan Africa are identified. These are medical, medico-moral, developmental (distinguishing between 'women in development' and gender and development perspectives), legal, ethical, and the rights discourse of groups living with HIV/AIDS and of African pressure groups. The analytical framework is that of discourse analysis as exemplified by Michel Foucault. The medical and medico-moral are identified as dominant. They shape the perceptions of the pandemic, our responses to it, and to those living with HIV/AIDS. However, dissident activist voices are fracturing the dominant frameworks, and are mobilising a struggle for meaning around definitions of gender, rights, and development. PMID- 8426963 TI - Descriptive and normative ethics: class, context and confidentiality for mothers with HIV. AB - This descriptive analysis reports on how institutional imperatives and professional assessments define, constrain, and even override normative claims for confidentiality in health care for mothers with HIV/AIDS. The analysis draws upon interviews with seventy-two health, social work, administrative, research, and advocacy professionals who work with mothers and infants with HIV/AIDS in nine sites in the U.S. and Puerto Rico and upon three analytical focus groups involving multidisciplinary groups of practitioners, lawyers, social scientists, and ethicists. Decisions regarding confidentiality for mothers with HIV/AIDS are likely to be influenced by (1) the relative lack of authority over information flow granted by medical institutions and professionals to patients and (2) professionals' class-based assumptions about mothers' needs for confidentiality and the consequences of withholding or disclosing information. Two normative conclusions for confidentiality practice in health care are drawn from and rooted in the descriptive findings of the research. First, if confidentiality practice is to be an expression of patient autonomy and rights, according to the principles generally held in medical settings, patients' control over information must be increased. Second, to avoid discrimination in the practice of confidentiality, decisions should be based on individual circumstances, not stereotypical generalities about social class. PMID- 8426964 TI - Individual rights vs the public health: the problem of the Asian hepatitis B carriers in America. AB - The influx of Asians into the United States (both as refugees and immigrants) presented American public health authorities with a significant challenge because many of them were chronic hepatitis B carriers. The carriers posed a real, but undetermined level of risk of spreading the infection to the larger community. Of special importance was the danger that susceptible children would be infected in day-care centers, because the earlier the age of exposure, the greater the risk of developing chronic carriership; which in turn is associated with cirrhosis and cancer of the liver in later life. The Asian carriers presented both a health problem, and a moral dilemma. If the carrier situation became a subject of public debate, there was danger of a racist and discriminatory reaction to the newcomers. Faced with this problem, American public health authorities opted to protect the Asian carriers by assuring the medical profession and concerned members of the public that there was either no health risk, or the risk was easily controllable, despite the lack of evidence to support such positions. In their attempt to protect a vulnerable group from discrimination, they functioned as humanitarians, but abrogated their duty as scientists and thereby undermined the long-term credibility of those charged with protecting the public health. Thus, despite achieving some immediate benefits for a defenseless group, the price they paid for that protection was too high. PMID- 8426965 TI - The quality of social roles as predictors of morbidity and mortality. AB - This study uses a longitudinal design to examine the health effects of the quality of social roles among a cohort of women and men. The specific roles of interest are the spouse role, the parent role, and the worker role. The cumulative and interactive effects of role characteristics and role satisfactions and stresses are also examined. The study population was randomly selected from among members of a large HMO and were part of a household interview conducted in 1970-71. Medical records for the 2 years prior to the interview and for 15 years after the interview for the cohort members are linked with the survey data. The findings show that for women, particular qualities of the marital and work roles are predictive of subsequent morbidity and mortality. For married women, equality in decision making and companionship in marriage are protective against death. None of the marriage characteristics are predictive of health outcomes among men. Work support is protective against death, malignancy, and stroke among employed women, and work stress increases the risk of ischemic heart disease among employed men. None of the parental role characteristics were significant predictors of health outcomes for men or women. No interactions were found. PMID- 8426966 TI - Managerial style and health promotion programs. AB - Organizational correlates of worksite health promotion programs were isolated and interpreted within a diffusion of innovation framework. A sample of managers from California (U.S.A.) 500 organizations were interviewed via telephone on their corporate management styles and health care strategies. Organizational management style was found to be related to prevalence of health promotion programs and future plans for health promotion programs. Specifically, this study found that organizations with democratic management styles are more likely to plan, adopt, and/or implement worksite health promotion programs when compared to organizations with authoritarian management styles. An additional contribution of this study was the development and validation of the Organizational Management Style (OMS) scale. These results have important theoretical and practical implications. For example, these findings explain why some organizations are more or less likely to adopt health promotion programs. Both diffusion of innovation and social control explanations are used to interpret the results. PMID- 8426967 TI - The Islamic dialogue with African traditional religion: divination and health care. AB - In rural Africa delivering better health care is more complicated than simply offering good medical services; it must also take into account peoples' religious orientation and beliefs. Here the non-material causes of illness are at least as important as the biological or material and, in many places, one can only determine the exact nature of the problem and its corresponding remedy after the fact, through a process involving divination and sacrifice. In northern Ghana, by adapting to traditional methods of divination, Islam is gradually introducing a future perspective and expanding the possibilities of preventative action. By so doing it is bringing about a religio-epistemological transformation that is, among other things, helping people to understand and make better use of Western bio-medicine and primary health care programs. The author argues for a similar adaptation and dialogue between African traditional beliefs and Western medical institutions. PMID- 8426968 TI - The power of compassion: truth-telling among American doctors in the care of dying patients. AB - The perceptions of American doctors about their practice regarding truth-telling in the care of dying patients were examined based on semi-structured interviews with 32 physicians in a teaching hospital. The doctors inform patients of their disease using three basic styles; 'telling what patients want to know', 'telling what patients need to know' and 'translating information into terms that patients can take'. These styles are supported by five basic normative principles; 'respect the truth', 'patients rights', 'doctors' duty to inform', 'preserve hope' and 'individual contract between patients and doctors'. These styles and principles suggest that physicians adhere to the recent trends of American medical ethics based on informed consent doctrine, and give the impression that patients have control over obtaining information. But close analysis of their accounts shows that physicians still hold power to control information through their management of the information-giving process. The styles and principles are flexibly interpreted and selectively used in the process so that they facilitate a discourse which justifies, rather than eliminates, the information control. Clinical contexts of information control are analyzed by examining dissimilar manners of providing information about treatment as opposed to prognosis. Physicians give less, and vaguer information about prognosis, citing its uncertainty and lesser relevance to future actions as reasons. Information about treatment is more readily shared in order to counterbalance the negative impact of the news on patients. The analysis reveals that the way doctors control information is closely related to the way they handle aspects of the reality of clinical practice, such as physicians' own emotional coping, institutional and legal constraints, and power relationships among patients, doctors and other care givers. Situating the findings in the historical context of normative discourse in American medicine, discussion focuses on the issues of trust and power of doctors. The humanistic role of the doctor, although suppressed in the currently dominant, contractual ethical framework, is still powerful in doctors' narratives. It expresses doctors' commitment to patients while preserving their authority. Implications of the individualistic approach to the doctor-patient relationship are also discussed. PMID- 8426969 TI - Post-empiricism and psychiatry: meaning and methodology in cross-cultural research. AB - Methodological disputes in psychiatry have usually been framed in terms of a positivist-interpretivist paradigm conflict. While traditional cross-cultural psychiatry does indeed endorse a positivist medical model the author argues that a post-empiricist philosophy of science is best able to 'ground' the 'new' cross cultural approach. PMID- 8426970 TI - Living longer but doing worse: assessing health status in elderly persons at two points in time in Manitoba, Canada, 1971 and 1983. AB - Comparisons of the health status of 1971 and 1983 samples of elderly persons in Manitoba, Canada suggest that while elderly individuals were living longer in 1983, their health was poorer. This was true in both age- and sex-specific comparisons and in comparisons made of individuals in the two samples who were relatively close to death. 'Compression of morbidity' has not taken place. Elderly individuals in the 1983 sample were in poorer health whether judged by functional status (ability to perform activities of daily living), number of different health problems reported, mental status or the rate of hospitalization for serious co-morbid disease. We estimate a 29% increase in the number of elderly persons resident in Manitoba over the 12 year period studied, but a 73% increase in the number of elderly who were in poor health. PMID- 8426971 TI - Colostrum and ideas about bad milk: a case study from Guinea-Bissau. AB - The study aims to explore ideas about bad milk found among women in Guinea Bissau. Interviews were held with 20 elderly knowledgeable rural women. Interinformant agreement was high within each ethnic group studied. All the informants recognized colostrum but disliked its consistency. Depending on ethnical background, it was considered good, of no special value or harmful to the newborn baby. Further, all the informants held that mature breast milk could turn bad, e.g. in case of mother's sickness or adultery. Suspected bad milk can be diagnosed by putting an ant into it to observe if it dies. The condition of bad milk can be treated by various procedures. The findings are discussed in relation to similar ideas existing in other societies and to views on the quality of maternal milk held in the industrialized countries. It is proposed that the idea of producing bad milk may be an important determinant of breastfeeding performance generally. Restrictions imposed on the breastfeeding woman, with the intention of producing healthier breast milk, may actually contribute to a decline in breastfeeding. PMID- 8426972 TI - Type A behavior: contextual effects within a southern black community. AB - The relationships among Type A behavior (assessed by the Framingham Type A scale), reported physical symptoms, and blood pressure were examined in a study of an African-American community in the rural southern U.S.A. The study was designed to determine: (a) if the cultural context of a black community altered the definition of Type A behavior; and (b) if the effects of Type A behavior were modified by this cultural context, as well as by socioeconomic and social structural variables. It was found that the patterning of traits characteristic of Type A behavior was different in the black (vs published studies of the majority) community, and that subscales of Type A behavior in turn had different effects on health variables than those observed in published studies. The health effects of these subscales were also moderated by socioeconomic and social structural variables. Future research should examine more closely how the definition of the Type A behavior pattern, and its effects, are modified by social and cultural context. PMID- 8426973 TI - Women's work and infant care in the Philippines. AB - Using data from a survey of 3000 Filipino infants and mothers, we analyze the implications of having more than one preschooler on several work outcomes, as well as the effect of availability of substitute care givers. The work outcomes of interest are those thought to be least compatible with infant care: work outside the home, wage work remunerated on a time basis, and long working hours. Our findings suggest that having multiple preschoolers only makes a difference for mothers whose household income is below the median: it increases their likelihood of working outside the home. Grandmothers and paid helpers consistently increase the likelihood of the various work outcomes and increase hours worked. Our findings suggest that urban residence, an indicator of work opportunity structure, increases the likelihood of wage-time work and is associated with longer working hours. PMID- 8426974 TI - Intrahousehold allocation of resources in larger and smaller Mexican households. AB - This paper focuses on aspects of intrahousehold allocation of resources. It suggests that there is a first step involved in understanding household decisions as to allocation or misallocation of food within the household, which is that of understanding intrahousehold allocation of income. In this study, carried out in northwestern Mexico, what has been reported in the literature as examples of unequal access to food, particularly in larger households, may better be considered examples of lack of access on the part of purchasers of household food to all of the income which comes into the household. PMID- 8426975 TI - Women's views of ultrasonography. A comparison of women's experiences of antenatal ultrasound screening with cerebral ultrasound of their newborn infant. AB - Ultrasound screening is now a routine procedure which forms part of antenatal care provision. Within this routine context ultrasound technology has been found to be generally acceptable and indeed is positively demanded by many women. This paper raises the question whether the routine presentation of ultrasound implicitly conveys the message that its use in antenatal care is both valuable and safe. It examines women's views of ultrasound technology beyond a routine context. In study designed to examine women's reactions to cerebral ultrasound on their normal term infants mothers were asked their views and knowledge of ultrasound and a comparison with their antenatal experience of ultrasound was elicited. A generalized concern about ultrasound techniques was found to underlie many of the women's comments. This raised questions concerning the current practice in the presentation of ultrasound to women attending for antenatal care. PMID- 8426976 TI - Socio-cultural determinants of child mortality in southern Peru: including some methodological considerations. AB - Among Amerindian children living at high altitude in the Andes in southern Peru, high child mortality rates have been reported in the literature, especially in the perinatal and neonatal period. We compared mortality rates in children calculated from retrospective survey data in 86 rural families from 2 Aymara and 3 Quechua peasant communities living at the same level of altitude (3825 m) in southern Peru. Relations between land tenure, socio-cultural factors and child mortality were studied, and methodological considerations in this field of interest are discussed. Checks on consistency of empirical data showed evidence for underreporting of neonatal female deaths with birth order 3 and more. Perinatal (124 vs 34 per 1000 births) and infant mortality (223 vs 111 per 1000 live births) was significantly higher in Aymara compared with Quechua children, but no difference was found after the first year of life. A short pregnancy interval was associated with an elevated perinatal and infant mortality rate, and a similar albeit insignificant association was found with increased maternal age. Amount of land owned and birth order were not related with child mortality. Although levels of maternal education are generally low in both cultures, a consistent decline in infant and child mortality was found with the amount of years mothers had attended school. However, the results suggest a U-shaped relationship between the amount of years of parental education and perinatal mortality in offspring. Late fetal and early neonatal mortality were particularly high in one Aymara community where mothers were found to have more years of education. Infanticide, a known phenomenon in the highlands of the Andes, is discussed in relation with the findings of the study. Although maternal and child health services are utilized by the majority of families in 4 of 5 study communities, 43 of 51 mothers under the age of 45 years reported that they delivered their last baby in the absence of traditional midwives or official medical supervision. PMID- 8426977 TI - Pregnancy and childbirth among the Amish. AB - This study examined Amish patterns of perinatal health care utilization from the perspective of Amish women and local health care providers in Geauga County, Ohio. Participant observation and interviews with health care providers and 15 Amish women yielded data on perinatal beliefs and utilization patterns for 76 pregnancies. While local health care providers regard the Amish as suboptimally utilizing prenatal care, this study found a consistent pattern of health seeking behavior. In the absence of symptoms perceived to be serious, Amish women initiated prenatal care earlier for first pregnancies and progressively later with increasing parity. Amish women's perinatal health care utilization must be seen within the context of barriers of transportation, cost, and child care needs. The Amish do not automatically reject medical technology, but select those aspects that are congruent with and that will support and maintain their way of life. Further, despite outward appearances of homogeneity, Amish women display individual variability in responding to pregnancy and childbirth. PMID- 8426978 TI - Factors associated with high-quantity prescriptions of benzodiazepines in Sweden. AB - An important aspect, when discussing the prescribing and use of psychotropics, is the amount of drug prescribed at each visit. In this study the Swedish Diagnosis and Therapy Survey was used in order to analyze high-quantity prescriptions of benzodiazepines. The total amount of benzodiazepines prescribed--measured as defined daily doses (DDD)--was calculated for each visit. Prescriptions with quantities equal to or greater than the 90th percentile applied on the distribution of prescribed DDD per visit were defined as high-quantity prescriptions. Using this definition, prescriptions on 200 DDD or more--14.9% of all--were classified as high-quantity prescriptions. In the analysis the proportions of high-quantity prescription in different subgroups were compared. The study showed that there was a strong relationship between age of the patient and high-quantity prescriptions while the sex of the patient was of minor importance. Doctors specializing in internal medicine and psychiatrists prescribed high-quantity prescriptions to a greater extent than other doctors but differences with regard to the doctor's ages were small. Patients with sleeping disturbances obtained high-quantity prescriptions to a greater extent than other patients while patients with nervous problems obtained fewer. Patients with new prescriptions on benzodiazepines obtained high-quantity prescriptions to a lesser extent than patients making repeat visits. In addition the study showed that it was not as common with high-quantity prescriptions in the three major cities and in the most sparsely populated communities as in mid-sized communities. PMID- 8426979 TI - Parental social support and child behaviour problems in different populations and socioeconomic groups: a methodological study. AB - In many research projects there is a need to apply a multifactorial approach. Practical reasons often make it necessary to use instruments that are not too time-consuming for each factor to be investigated. The Interview Schedule of Social Integration (ISSI) is proposed by Unden and Orth-Gomer as a suitable tool in measuring social support. It is increasingly used in Sweden. To further investigate its usefulness in measuring parental social support of importance for the children, an examination of the relation between the ISSI variables and Child Behaviour as well as Parental Psychopathology was undertaken, based on data from four studies: Psychosocial Factors and Child Diabetes, Single Parent Families in a child guidance clinic, a Clinical Child Psychiatric Ward Group and an Epidemiological Study of a Normal Swedish Population. The findings were that social support measured by ISSI had the expected associations to child behaviour as well as to parental psychopathology in the higher socioeconomic groups but not in the lower ones. The interpretation is made that ISSI does not measure qualitative aspects of social support but the sense of satisfaction with it. In the higher socioeconomic groups the sense of satisfaction may be a fairly good approximation of the actual quality even if it is probably confounded with personality trait factors. The same approximation cannot be made in lower socioeconomic groups and in psychiatric populations. In these cases there is a need to pay more attention to the actual quality of the social network. PMID- 8426980 TI - Dental care use by U.S. veterans eligible for VA care. AB - Military veterans eligible for dental care in U.S. Department of Veterans Affairs (VA) facilities cooperated for a mailed survey about their dental care utilization. Subjects were selected because of their eligibility for continuing dental care in VA facilities at no monetary cost. However, only 48% reported the VA as their only or primary source of dental care; this allowed us the opportunity to compare dental care frequency by those who received dental care at no monetary cost with those who did not, as well as measure delivery system effects on dental care use. Consequently, we tested respondent-level and delivery system-level hypotheses regarding determinants of veterans' dental care use. Predisposing characteristics (dentate status, usual reason for dental visits, and the importance placed on dental care and oral health) were the strongest determinants of interval since last dental visit. Enabling determinants (current source of dental care, and having a regular source of care) were also significant, but measures of need for dental care (perceived oral health and perceived need for treatment) were not. More recent dental care use by veterans who used the VA delivery system as their source of dental care, even with dental care payment source and other determinants accounted for, suggests that the VA delivery system may have promoted more regular use compared to other systems. PMID- 8426981 TI - Mothers brought dead: an enquiry into causes of delay. AB - In a 10 year period (1981-1990) 118 pregnant or recently delivered women were dead when they reached hospital. Relevant history with emphasis on the events of the previous 24 hr and the reasons for delay in reaching the hospital were obtained from the relatives accompanying the patient. The probable cause of death was determined on the basis of history only. Autopsy was not carried out on any case. The causes of delay were economic factors in 42, socio-cultural factors in 39 and inadequate maternal services in 24 cases. In 13 cases the reasons for delay could not be established. The medical problems were either preventable or treatable if managed in time. A combination of economic, social and cultural factors played a more significant role in these deaths than medical causes. PMID- 8426982 TI - Perception of diarrhoeal diseases among mothers and mothers-to-be: implications for health education in Saudi Arabia. AB - The prevalent beliefs and therapeutic preferences for diarrhoea among mothers and unmarried high school girls from an urban area in Eastern Saudi Arabia were investigated. Knowledge regarding dehydration and malnutrition as a complication of diarrhoea was far from optimum in the studied groups. It was encouraging to note that a large proportion of mothers and students were not in favor of restricting food and fluids during the illness. A long list of home remedies for diarrhoea was provided by them. Two-thirds of the students opted for Oral Rehydration Therapy as the best treatment in diarrhoea whereas mothers who were in general less educated, preferred antidiarrheal liquids and injections. Channels of communication to health educate mothers and mothers-to-be are suggested. PMID- 8426983 TI - The neurobiology of the association of women's job-related conflict with depressive symptomatology. PMID- 8426984 TI - [Computed tomography of the pleura and the parenchyma in workers exposed to asbestos]. PMID- 8426985 TI - [An aneurysm of the ischiadic artery]. PMID- 8426986 TI - [Pre-angioplastic detection of a hemodynamically active iliac artery stenosis in an implanted femoro-femoral bypass using an intra-arterial injection of prostaglandin E1]. PMID- 8426987 TI - [Radiation exposure to patients in computed tomographic studies]. PMID- 8426988 TI - [Radiation exposure and risk in nuclear medicine and x-ray diagnosis]. PMID- 8426989 TI - [Superselective placement of coaxial catheters in truncal arteries. Experience with the 18-catheter tracker]. PMID- 8426990 TI - Pathogenesis of colorectal cancer. AB - This article takes the reader from colorectal lumen to the intracellular environment in an effort to explain the processes of tumorigenesis in colorectal cancer. The molecular changes seen in colorectal cancer development also are discussed, along with the potential for genetic manipulation and diagnostic molecular techniques. An era of major advances in understanding the pathogenesis of colorectal cancer has begun. PMID- 8426991 TI - Principles of surgical resection. Influence of surgical technique on treatment outcome. AB - In the absence of curative medical therapy, surgical resection remains the cornerstone of treatment for patients with colorectal carcinoma. A thorough knowledge of colon and rectal anatomy is crucial for the formulation of an effective operative strategy. There are certain technical factors under the control of the surgeon that may have prognostic significance for the patient. These include the length of the distal margin of resection, the use of intraluminal cytotoxic solutions to reduce the viability of exfoliated cancer cells, and the technique of colon anastomosis. Curative resections should include removal of the lymphatic drainage of the tumor-bearing segment of colon. When there is adjacent organ invasion by the colonic primary, en block resection of the entire tumor mass with adequate margins is the procedure of choice. Prophylactic oophorectomy in women with colon carcinoma remains controversial. The effects of perioperative transfusion on tumor behavior remain unclear. Blood transfusions should be administered only when there is a specific medical necessity. PMID- 8426992 TI - Extended resection for carcinoma of colon and rectum. AB - Although surgery has been the mainstay of treatment for patients with colorectal carcinoma for more than a century, debate continues regarding the appropriate magnitude of operation for optimal survival. Invasion of contiguous organs is a legitimate indication for extended en bloc resection, including pelvic exenteration, in appropriately selected individuals. Extended lymphadenectomy, especially in resections for carcinoma of the rectum, is being reexamined with renewed enthusiasm. Improved perioperative care has permitted performance of more aggressive operative intervention, with improved cure rates for patients with colorectal neoplasms. PMID- 8426993 TI - Sphincter-saving alternatives for treatment of adenocarcinoma involving distal rectum. AB - In reviewing the results achieved with local treatment of carcinoma of the rectum, it becomes apparent that no single approach to the treatment of carcinoma of the rectum is optimal for all patients. As is true in patients with carcinoma of the breast and other malignant lesions, selection of the appropriate treatment alternatives for a patient with carcinoma confined to the distal rectum requires accurate preoperative staging of the disease. Physicians interested in treating patients with carcinoma of the rectum should have the full spectrum of treatment alternatives available. PMID- 8426994 TI - Management of recurrent and metastatic colorectal carcinoma. AB - When metastatic or recurrent disease from colorectal carcinoma is detected, the surgeon must decide whether a patient is a candidate for resection. Although long term survival after resection is not optimal, the relegation of patients to nonresective treatment means denying them the only chance for cure currently available. When isolated disease involving the liver, lung, or region of the primary carcinoma is documented, curative resection must be considered. Symptomatic patients may also obtain maximal palliation from resection, diversion, or a bypass procedure. Chemotherapy for the treatment of recurrent disease is palliative and probably should be considered only within clinical trials. Future alternative methods of treatment or new chemotherapeutic regimens need to be studied to improve survival and quality of life. PMID- 8426995 TI - Radiation therapy for colorectal cancer. AB - The role of radiation therapy in the management of colorectal cancer has become more clearly defined as the number of clinical studies has grown. It is now evident that radiation is capable of sterilizing subclinical deposits of cancer at doses tolerable by adjacent normal tissues, and to a lesser extent, these doses can control more bulky cancers. The integration of radiation and chemotherapy has already led to some improvement in survival rates in the adjuvant treatment of rectal cancer. The further development of such combinations seems likely to improve tumor control and survival rates in many stages of cancer. In the next decade, it is also likely that there will be refinement of the use of radiation through better understanding of the biology of colorectal cancer, perhaps supplemented by the development of predictive assays that can guide both the selection of patients for treatment and the choice of the most effective radiation schedule. PMID- 8426996 TI - Screening strategies for colorectal cancer. AB - The efficacy of screening for colorectal cancer has not been established. Policy making organizations differ in recommendations for asymptomatic and high-risk groups because of the inadequacy of current evidence. A critical appraisal of the current evidence for screening for colorectal cancer and a discussion of the aims and pitfalls of screening programs are presented. PMID- 8426997 TI - Management of malignant colorectal polyps. AB - The optimal management of patients with adenomatous polyps that contain invasive adenocarcinoma remains controversial. The independent factors of margins of resection, level of invasion, differentiation, grade, and vascular invasion are examined as prognostic indicators for outcome. The literature is reviewed with regard to the management of patients with polyp-containing invasive adenocarcinoma with standard operative resection versus endoscopic treatment alone. PMID- 8426998 TI - Preoperative evaluation and postoperative surveillance for patients with colorectal carcinoma. AB - As the understanding of the biology of carcinoma of the colon and rectum increases and the effectiveness of adjuvant therapy for patients with such carcinoma improves, preoperative staging of disease has assumed an increasingly important role. The rationale and specifics of preoperative evaluation and the role, rationale, and controversies regarding postoperative surveillance after curative resection of colorectal carcinoma are discussed. PMID- 8426999 TI - Biochemical markers for colorectal cancer. Diagnostic and therapeutic implications. AB - The development of the understanding of oncogenes, tumor suppressor genes, and signal transduction has provided a significant advance in the concepts of the mechanisms of carcinogenesis in the colon and rectum. The tools provided by the molecular geneticist and the immunobiologist may yield powerful new techniques for screening individuals at risk, for identifying those patients with biologically more aggressive tumors, for developing novel therapies targeted directly at tumor cells, and for providing the means for more sensitive and specific detection of recurrence of disease. PMID- 8427000 TI - Reconstruction of the hepatic vein in reduced size hepatic transplantation. AB - Reconstruction of the hepatic vein (HV) is not required in size-matched orthotopic liver transplantation (OLT) because the vena cava (VC) is replaced. In reduced size OLT, used for providing small livers for children, the HV is often implanted directly. Grafts obtained from a split liver in which the right lobe is used for a second recipient or from a live donor must be implanted without the VC. To evaluate the occurrence of outflow complications and their prevention, we have reviewed our experience with 72 left sided reduced grafts in children. Between July 1985 and November 1990, 93 reduced grafts were performed. Twenty-one were right lobe grafts with orthotopic replacement of the VC. Seventy-two were left grafts comprising 28 full left lobes and 44 lateral segments. Grafts were obtained from reduction of a cadaver liver in 39, from the left lobe of a split liver in 21 and from a live donor in 12. Of the left grafts, 47 were implanted with preservation of the recipient VC. Overall, HV obstruction occurred in 12 patients. Obstruction occurred acutely in three patients, causing graft failure and death in two and was repaired successfully in one patient. Chronic HV obstruction was documented in three patients with ascites and graft enlargement requiring retransplantation. This complication occurred in five of 25 patients with VC, six of 18 with end to end HV anastomosis, one of 18 with end to side implantation of HV and zero of 15 using a triangular anastomosis (p = 0.05). Outflow obstruction has not received adequate attention in descriptions of reduced-size OLT. Marked hepatic swelling and fluid retention that occur after reduced size hepatic transplantation may be the result of incomplete HV obstruction. In this series, end to end anastomosis of the HV resulted in a high frequency of outflow obstruction. This was prevented when anastomoses were designed to allow the graft to rest comfortably in the hepatic fossa after abdominal closure. PMID- 8427001 TI - Expanded polytetrafluoroethylene patch versus polypropylene mesh for the repair of contaminated defects of the abdominal wall. AB - Contaminated defects of the abdominal wall continue to be a significant problem for patients and surgeons. The lack of sufficient tissue may require the insertion of a prosthetic material. Polypropylene (PP) mesh is still the most widely used material for this purpose, although the propensity to induce extensive visceral adhesions and erosion of the skin or intestine is a well-known drawback. Expanded polytetrafluoroethylene (PTFE) patch has better mechanical properties and has a low potential for infection. Therefore, we used expanded PTFE patch to repair contaminated abdominal wall defects in three patients. In one patient, the postoperative course was uneventful. In the other two patients, the patch had to be removed for ongoing wound sepsis and because the patch disintegrated. In an experimental study, contaminated abdominal wall defects created in Wistar rats were repaired with expanded PTFE patch (PTFE group, n = 21) or PP (PP group, n = 21). Wound infection occurred in 16 rats in the PTFE group and in 14 rats in the PP group. Two rats in each group died. Two rats in the PTFE group died as a result of peritonitis, one rat in the PP group died as a result of ileus and one as a result of peritonitis. Incisional hernia was found to be significantly more frequent in the PTFE group (n = 13) than in the PP group (n = 3). Fistula formation was only found in three rats in the PP group. Adhesion formation was more pronounced in rats in the PP group. It is concluded that the expanded PTFE is unsuitable for the reconstruction of contaminated abdominal wall defects and that PP mesh is more suitable, although this material has a high risk of complications. PMID- 8427002 TI - Pre-preparation of the sterile instrument table for emergency cesarean section. AB - Cesarean section instrument tables are often not prepared in advance because of concern of contamination risk. The Association of Operating Room Nurses Standards decries the use of pre-preparation of surgical instrument tables because of this risk, although there are no scientific data to support this claim. We evaluated the contamination risk of pre-preparation of surgical instrument tables, prolonged table coverage and table uncovering using a specific technique referred to as the "sardine can roll." Colony counts were positive in only seven of 180 cultures (< or = 15 colonies per plate in each instance) from six tables evaluated after prolonged coverage or uncovering, or both. These data suggest that contamination risk is slight for the uncovering technique described herein and advance table preparation (24 hours or less, never recovered) is a reasonable clinical option in units in which table preparation reduces response time in emergent clinical situations, such as cesarean section for acute fetal distress. PMID- 8427003 TI - Experimental basis and clinical application of extended highly selective vagotomy for duodenal ulcer. AB - An ideal operation for a benign disease, such as duodenal ulcer, must be effective, relatively free of serious postoperative complications and standardized for effective performance. Highly selective vagotomy, an operation known to have few side effects, has not been adopted by many surgeons because of the concerns about postoperative recurrent ulcers, as well as by concerns about the technical performance of the operation. Recurrent ulcers may be the result of incomplete denervation of the parietal cell mass; based upon the results of intraoperative testing, we suspected that previously unrecognized sites supplied preganglionic vagus nerves to the parietal cell mass. Nerve tracing studies identified four areas in the stomach that are not adequately denervated in the "standard" highly selective vagotomy. Based on these data, we defined and performed "extended highly selective vagotomy" upon 180 patients with ulcers. Herein, the results of follow-up examinations of 113 patients for a mean of at least 4.5 years postoperatively are reported. Recurrent duodenal ulcers occurred in 2.9 percent (two of 69) after extended vagotomy alone, and in none of 44 in whom a drainage procedure was used; the overall recurrence rate was 1.8 percent. Serious postoperative complaints were found in one patient in each group. The use of extended highly selective vagotomy for duodenal ulcer has a rational experimental basis and acceptable clinical results. Pyloroplasty, in conjunction with extended highly selective vagotomy, does not increase the incidence of serious postoperative side effects. PMID- 8427004 TI - Indications and limitations of percutaneous cholecystostomy for acute cholecystitis. AB - Percutaneous cholecystostomy (PC) was used as an alternative to operative therapy in 21 elderly or critically ill patients with suspected acute cholecystitis. All had associated disabling diseases, ten (48 percent) were older than 65 years, nine (43 percent) were in the intensive care unit and eight (38 percent) were recovering from recent operations. Among the 21 patients, 18 had cholecystitis (eight calculous and ten acalculous); in three patients, the procedure was only diagnostic. In 16 of 18 patients with acute cholecystitis, immediate relief of symptoms and significant improvement of laboratory signs of cholecystitis occurred. Persistent signs of peritonitis and uncertainty of diagnosis led to cholecystectomy without complication in one patient. Colonic perforation, as a result of technical complications, necessitated laparotomy in another patient. No other complication of PC was noted. Mortality rate of a patient with cholecystitis, treated by PC, was 5.5 percent (one of 18). Among ten patients with acalculous cholecystitis, only one patient underwent cholecystectomy because of a direct complication of PC. The other nine patients are alive and symptom free with an intact gallbladder after a mean follow-up period of 16 months. Among eight patients with calculous cholecystitis, four later underwent cholecystectomy, three died from underlying disease, and one patient had stones extracted percutaneously. These results indicate that PC is an effective temporary measure in elderly or critically ill patients with acute cholecystitis and, under close clinical supervision, a safe alternative to surgical intervention. In patients with acalculous cholecystitis, PC can be used as an immediate and definitive therapy, and cholecystectomy can be avoided. PMID- 8427005 TI - Chilaiditi's syndrome as a surgical and nonsurgical problem. AB - Hepatodiaphragmatic interposition of the intestine is a rare anomaly (0.025 to 0.28 percent of the general population) described by Chilaiditi in 1911 and often believed to be of irrelevant clinical interest. To the contrary, recent studies stated that this syndrome is a potential source of abdominal problems requiring emergency or elective operation. From a retrospective analysis of records since 1976, four instances of Chilaiditi's syndrome have been found (three males and one female). Interposition of the proximal transverse colon was found in three patients and the small intestine in one patient. The findings of plain roentgenograms of the chest were determinants for diagnosis in three patients. In one patient, a barium meal was given to obtain a better definition. Two patients were admitted for malignant neoplasms (metastatic carcinoma of the breast, carcinoma of the gastrointestinal tract and cirrhosis of the liver) and died within a few months. The other two patients complained of abdominal pain. Patient No. 4 had gastric volvulus. Chilaiditi's syndrome was diagnosed intraoperatively in that patient and a surgical treatment with hepatopexy was performed, by suturing the falciform, the coronaria ligament and the anterior margin of the liver to the diaphragm with interrupted absorbable stitches. After a two year follow-up evaluation, this patient is as well as the patient who underwent medical therapy. Volvulus of the stomach, as was found in Patient No. 4, is an unusual condition and, to our knowledge, the patient is the second reported instance. PMID- 8427006 TI - Radiotelemetric fetal monitoring during and after open fetal operation. AB - Diagnosis and treatment of fetal disease processes has advanced dramatically, but the ability to monitor the fetus in utero remains rudimentary. Open fetal operation provides a unique opportunity to correct life-threatening fetal abnormalities, but it also places the fetus at risk. Continuous intraoperative and post-operative fetal electrocardiographic monitoring may decrease the risk by optimizing fetal and maternal perioperative management. We tested an implantable radiotelemetry device that allowed continuous intraoperative and postoperative fetal electrocardiographic and temperature monitoring. The radiotelemetry device was placed subcutaneously in four fetal sheep at 100 to 112 days' gestation (term equals 145 days). During and after the fetal operation, the monitor reliably and continuously transmitted the analog fetal electrocardiogram and temperature. We then tested a similar device during human fetal operations on both an acute and a chronic basis. The radiotelemetry device permits continuous and reliable fetal monitoring during and after fetal operation and we now consider it essential to the operation. Its use should facilitate optimal fetal and maternal management and improve survival of the fetus. PMID- 8427007 TI - The effect of anabolic steroids on ameliorating the adverse effects of chronic corticosteroids on intestinal anastomotic healing in rabbits. AB - The ability of anabolic steroids to reverse the deleterious effects of chronic corticosteroids on intestinal anastomotic healing was studied in 32 rabbits. Rabbits received once daily intramuscular injections of saline solution (controls), 0.1 milligram per kilogram of dexamethasone, dexamethasone plus low dose anabolic steroids (2 milligrams per kilogram of nandrolone decanoate) or dexamethasone plus high-dose anabolic steroid (10 milligrams per kilogram of nandrolone decanoate). Two weeks later, the rabbits underwent small intestinal (SI) or large intestinal (LI) anastomoses. Postoperatively, injections were continued as before with the addition of high-dose anabolic steroids (20 milligrams per kilogram of nandrolone decanoate) to one-half of the dexamethasone only group. Seven days postoperatively, the rabbits had in situ assessment of anastomotic bursting pressure (ABP) and histologic examination using a modified Ehrlich/Hunt scale. Results demonstrated that dexamethasone severely impaired the healing of SI and LI anastomoses, with decreases in ABP to almost 50 percent of normal values and with similar reduction in histologic parameters. High-dose anabolic steroids reversed this inhibitory effect when initiated preoperatively or post-operatively, with increase in ABP to 72 percent of normal for SI anastomoses and 83 percent of normal for LI. No adverse effect of anabolic steroid administration was evident. PMID- 8427008 TI - Optimal cosmetic autogenous reconstruction with modified radical mastectomy. PMID- 8427009 TI - Surgical considerations in the short bowel syndrome. AB - None of the current surgical alternatives for the short bowel syndrome is sufficiently safe and effective to be used routinely. Surgical therapy should be considered only in selected patients to achieve specific results. Patients with dilated intestinal segments and stasis may benefit from intestinal tapering and lengthening. The results of intestinal transplantation in animals have improved and justify clinical attempts, but recent experience in humans has been disappointing. Growing neomucosa has not been shown to increase absorption, but the patching technique may be useful in preserving intestinal length. Patients with sufficient absorptive area but rapid transit may benefit from colon interposition or intestinal valves. Thus, the surgical emphasis should continue to be prevention of intestinal resection and conservation of intestinal length when resection is required. PMID- 8427010 TI - Evaluation of [3H]paroxetine as an in vivo ligand for serotonin uptake sites: a quantitative autoradiographic study in the rat brain. AB - Paroxetine, a selective inhibitor of serotonin uptake and an antidepressant, was used in conjunction with quantitative ex vivo autoradiography to study the feasibility of imaging serotonin terminals in the living brain. Tritiated paroxetine was injected in the rat tail vein, and the brain was processed for quantitative autoradiography 3 hours later. Animals received either [3H]paroxetine alone (100 microCi/animal) or a mixture of labeled paroxetine (100 microCi) and an excess of unlabeled drug (0.5 or 2 mg/kg intravenously [i.v.]). Computerized image analysis of the resulting autoradiograms revealed high densities of radioactivity in brain regions known to contain high densities of serotonergic terminals and high specific binding of [3H] paroxetine in vitro, such as the raphe nuclei, interpeduncular nucleus, basolateral amygdala, substantia nigra, and some hypothalamic nuclei. Radioactivity uptake in these brain regions was effectively blocked (50-72%) by coadministration of excess unlabeled paroxetine. However, cortical and hippocampal binding of paroxetine in vivo was moderately high, in contrast to the relatively sparse serotonergic innervation in these regions. Only a relatively small proportion of cortical and hippocampal binding (20-40%) could be blocked by excess unlabeled paroxetine, indicating that most of the radioactivity in these regions is not associated with serotonin terminals or uptake sites. The usefulness of [3H]paroxetine as an in vivo ligand for imaging serotonin terminals in the human brain is limited by these nonserotonergic binding sites. PMID- 8427011 TI - The cytochromes P-450 are not involved in the modulation of the N-methyl-D aspartate response by sigma ligands in the rat CA3 dorsal hippocampus. AB - Recent in vitro radioligand binding studies have shown that several cytochrome P 450 inhibitors can displace [3H] sigma ligands, suggesting that these ligands might bind to the cytochrome P-450 superfamily of enzymes. Using an in vivo electrophysiological model of extracellular recordings performed in the CA3 region of the rat dorsal hippocampus, we have previously shown that intravenous administration of low doses of several sigma ligands, such as 1,3-di(2-tolyl) guanidine (DTG), JO-1784, and (+)pentazocine potentiate the neuronal response induced by microiontophoretic applications of N-methyl-D-aspartate (NMDA) without affecting those induced by quisqualate and kainate, suggesting that they act as sigma agonists. Conversely, the sigma ligands haloperidol, (+)3-PPP, and BMY 14802, which have no effect by themselves on the NMDA response, prevent and suppress the potentiating effect of sigma agonists on the NMDA response, suggesting that they act as sigma antagonists. The present studies were undertaken to determine if cytochromes P-450 could be involved in the modulation of the NMDA response by sigma ligands. For this purpose, two cytochrome P-450 inhibitors, proadifen (SKF-525A) and piperonyl butoxide (PB), have been tested in our model. Unlike sigma agonists, at low doses, neither SKF-525A nor PB affected the NMDA response of CA3 dorsal hippocampus pyramidal neurons. Unlike sigma antagonists, neither of these drugs reversed or prevented the DTG-induced potentiation of the NMDA response. In addition, following high doses of SKF-525A or PB, sufficient to induce a complete inactivation of cytochromes P-450, DTG still potentiated the NMDA response.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427012 TI - Morphological and electrophysiological evidence for electrotonic coupling of rat dorsolateral septal nucleus neurons in vitro. AB - Intracellular injections of Lucifer Yellow were utilized to evaluate the incidence of dye-coupling among dorsolateral septal nucleus (DLSN) neurons recorded from slice preparations of adult rat septal nuclei. Twenty percent of single injections of Lucifer Yellow resulted in pairs of labeled neurons. These dye-coupled cells were morphologically heterogeneous and did not exhibit any morphological characteristics that could be used to distinguish them from non dye coupled neurons. The spatial separation of cell bodies and close apposition of dendrites within each pair indicated that the dye transfer site(s) were situated at dendrodendritic and/or dendrosomatic rather than somatosomatic junctions. The main axon of some dye-coupled neurons gave rise to intrinsic axon collaterals prior to exiting the nucleus indicating that these coupled neurons function as projection neurons as well as local circuit interneurons. Electrophysiological recordings of the passive membrane properties and spontaneous activity of individual dye-coupled neurons revealed no significant difference from non dye coupled cells in the DLSN. Some neurons exhibited spontaneously occurring fast potentials which presumably represent electrotonic potentials. These fast potentials were often tightly coupled with action potentials but could be distinguished from synaptic potentials by their shape and their lack of voltage dependent changes in amplitude. These morphological and supportive electrophysiological data provide the first indirect evidence for electrotonic coupling of dorsolateral septal neurons. The functional significance of this coupling may lie in the potential for synchronization of the output of the DLSN which could play an important role in the septal maintenance and modulation of hippocampal Theta rhythm. PMID- 8427013 TI - Effect of differential rearing on synapses and soma size in rat medial amygdaloid nucleus. AB - The effects of differential rearing on synaptic morphology and neuronal size were examined in the medial amygdaloid nucleus (MAN) of adult rats. Forty-day-old male rats were housed in one of three ways: individually (isolated condition, IC); with four males per cage (unisexual condition, UC); or with two males and two females per cage (social condition, SC). After 2 months, the animals were prepared for electron and light microscopy. Although there was no statistically significant difference in number of synapses per unit volume in MAN, the number of perforated (P) synapses, which are characterized by discontinuities in the postsynaptic density, were significantly greater in the UC and the SC than in the IC. The length of synaptic contact zone of P synapses was also longer in both the UC and the SC compared with the IC, whereas the length of nonperforated synapses was longer only in the SC. Soma area was also larger in the SC compared with the IC. These results demonstrate that exposure to different rearing conditions, in which the pheromonal environment can be substantially different, can induce striking morphological changes in both synapses and neurons in the MAN of adult rats. PMID- 8427014 TI - Ventral tegmental self-stimulation selectively induces opioid peptide release in rat CNS. AB - Intracranial self-stimulation (ICS) is thought to activate neuronal systems involved in processing natural reinforcing agents. Metabolic mapping studies have previously demonstrated a subset of CNS structures specifically engaged by ICS in animals receiving stimulation actively vs. passively. Since opiates are known to enhance ICS behavior and presumably its reinforcing properties, the current study addressed the question of the role of opioid peptides as mediators of ICS. Rats were trained on a fixed ration (FR) 20 schedule of responding maintained by ICS. Following response stabilization, rats were assigned either to an active or a corresponding yoked stimulation group at 1 of 2 schedules of reinforcement (i.e., FR1-YFR1, FR20-YFR20, or sedentary control), and opioid peptide release was inferred from in vivo receptor occupancy. Autoradiographic analyses identified 3 groups of structures. Treatment-induced alterations in occupancy were seen in the medial dorsal nucleus of the thalamus, basolateral amygdala, ventral pallidum, medial habenula, dorsal raphe, posterior hypothalamus, substantia nigra pars compacta, agranular preinsular cortex, and zona incerta. Depending upon the structure, peptide release was dependent upon stimulus contingency (active vs. yoked) and/or schedule (FR1 vs. FR20). Evidence for ICS-induced inhibition of peptide release was found in the habenula and preinsular cortex. Nine additional structures, all components of, or receiving projections from, the limbic system, revealed complex interactions between ICS treatment and the electrode side. Finally, a widespread ipsilateral increase in receptor binding was seen rostrally from the cingulate, olfactory tubercle, and nucleus accumbens, along the lateral hypothalamus and hippocampus, and extending caudally to the substantia nigra and ventral tegmentum. These later effects appear to be related to stimulation induced changes in blood flow and subsequent ligant presentation increases. Collectively, these data point towards the ability of rewarding brain stimulation to activate discrete neuronal opioid systems contingent upon specific behavioral as well as stimulus conditions. PMID- 8427015 TI - SCE frequencies induced by ethanol, tequila and brandy in mouse bone marrow cells in vivo. AB - The genotoxicity of ethanol, tequila and brandy was evaluated by scoring the frequency of sister chromatid exchanges (SCE) and determining the values of the average generation time (AGT). We studied four dosages of each substance i.p. inoculated into mice. The cytogenetic analysis was performed in bone marrow cells. The results showed that all three substances were weak genotoxicants. Tequila showed the strongest response followed by brandy and ethanol. None of them modified the cell proliferation kinetics as demonstrated by the AGT results. PMID- 8427016 TI - Impairment of long-term potentiation and learning following chronic lead exposure. AB - Chronic lead exposure during brain development is known to affect functions of the central nervous system. We exposed rats chronically to low levels of lead at different developmental stages in order to determine the most sensitive periods of exposure. Active avoidance learning and hippocampal long-term potentiation were tested in the same animals. If the exposure period comprised the prenatal and the early postnatal phase and was continued into adulthood, learning as well as long-term potentiation were impaired. Starting the exposure not before 16 days postnatally, however, neither affected learning nor hippocampal potentiation. These results reflect the higher vulnerability of the immature as compared to the mature hippocampus to lead-induced functional deficits. PMID- 8427017 TI - Altered behaviour in adult mice orally exposed to tri- and tetrachloroethylene as neonates. AB - Male NMRI mice were orally exposed to trichloroethylene (TRI, 50 and 290 mg/kg per day) and tetrachloroethylene (PER, 5 and 320 mg/kg per day) between days 10 and 16 postnatally. Spontaneous activity, observed as locomotion, rearing and total activity, was measured over three 20-min periods at an age of 17 and 60 days. Compared to controls, mice at 17 days of age were unaffected, while at 60 days of age mice exposed to PER showed changes in all three spontaneous motor activity variables, while TRI-exposed mice only were affected in rearing. This indicates a neonatal susceptibility of brain maturation to these chlorinated organic solvents in achieving long-lasting changes in adult behaviour. PMID- 8427018 TI - Popliteal lymph node response to procainamide and isoniazid. Role of beta naphthoflavone, phenobarbitone and S9-mix pretreatment. AB - The popliteal lymph node assay (PLNA) was proposed for the preclinical prediction of xenobiotics-induced autoimmune reactions in humans. Among the substances so far tested in this model, procainamide and isoniazid gave negative PLNA responses despite reports of lupus syndromes in man. To confirm the hypothesis that a metabolite instead of the parent molecule is involved, rats were pretreated with phenobarbital or beta-naphthoflavone, then injected with procainamide or isoniazid. In additional groups of animals, procainamide or isoniazid were injected together with S9-mix following various incubation times. Pretreated rats had a positive PLNA response when injected with procainamide, whereas preincubation with S9-mix resulted in a positive response to isoniazid. These results further support the validity of the PLNA. PMID- 8427019 TI - Effects of p-phenylenediamines and adriamycin on primary culture of rat skeletal muscle cells. AB - In vitro toxicity of p-phenylenediamine derivatives and adriamycin were examined using cultured rat myofibers prepared by the selective plating method, with slight modification. When the myofibers were cultured for 2 days with 100 microM N,N,N',N'-tetramethyl p-phenylenediamine, atrophy and/or swelling of the cells were observed, and the toxic effect was reduced as the number of N-methyl groups in the phenylenediamine molecule was decreased. Adriamycin, at concentrations of greater than 0.25 microM, caused cell injury. Decline of cellular creatine phosphokinase activities generally preceded the apparent morphological changes. The primary culture of rat myofibers responded sensitively to the agents which were myotoxic in vivo. PMID- 8427020 TI - Strain differences in susceptibility to di-2-ethylhexyl phthalate-induced testicular atrophy in mice. AB - It is well known that the oral administration of di-2-ethylhexyl phthalate (DEHP) produces severe seminiferous tubular atrophy in rats, but mice are far less sensitive. In the present experiment, we demonstrated that one strain of mice derived from ICR mice, Crj:CD-1, was very sensitive to DEHP-induced testicular injury. This atrophy was accompanied by lowered zinc concentration and lactate dehydrogenase isozyme X in the testis. This fact and the effect of genetic background will need to be considered for assessing the risk of testicular toxicity in men exposed to phthalate esters. PMID- 8427021 TI - Renal lesions induced in F344/DuCrj rats by 4-weeks oral administration of dimethylarsinic acid. AB - The nephrotoxicity of dimethylarsinic acid (cacodylic acid, DMA) was examined in male and female F344/DuCrj rats. DMA administered perorally at doses of 113, 85, and 57 mg/kg for 4 weeks produced dose-related decreases in body weight and survival rate in both sexes. Mortality was higher and appeared more quickly in females than in males. Histopathological findings in the kidney were proximal tubular degeneration and necrosis, as well as papillary necrosis, and hyperplasia of the epithelium covering the papillae. Since extensive proximal tubular necrosis was observed only in dead animals of both sexes, and not in survivors or the controls, it was therefore concluded that the main cause of death could be attributed to nephrotoxicity of DMA. The results thus show that DMA is nephrotoxic to both male and female rats. PMID- 8427022 TI - Role of lipid peroxidation in acetaminophen-induced hepatotoxicity: comparison with carbon tetrachloride. AB - The effect of acetaminophen on lipid peroxidation in vivo and in vitro was studied in rat liver and the data were compared with those with carbon tetrachloride. Carbon tetrachloride increased diene conjugates in vivo and thiobarbituric acid reactive substance production in vitro in the liver microsomal incubation. These changes were further enhanced by ethanol that has previously been shown to increase carbon tetrachloride-induced hepatotoxicity. On the other hand, acetaminophen did not increase diene conjugates in vivo and inhibited thiobarbituric acid reactive substance production in vitro. These effects were minimally affected by ethanol which has previously been shown to inhibit acetaminophen-induced hepatotoxicity. Thus, lipid peroxidation may play a minimal role in acetaminophen-induced hepatotoxicity in contrast with carbon tetrachloride-induced hepatotoxicity. PMID- 8427023 TI - Protective effect of calmodulin inhibitors against acute cyanide-induced lethality and convulsions in mice. AB - The ED50 value of cyanide as measured by induction of convulsions (tonic seizure) was significantly increased by 80% or 69% when trifluoperazine (TFP) or chlorpromazine (CHP), a specific calmodulin inhibitor was preinjected intracerebroventricularly (i.v.t.) at a dose of 0.09 mumol/body of mice. However, the ED50 of cyanide was not significantly changed by the same dose of promethazine (PMZ), another calmodulin inhibitor. Since it is known that the inhibitory effect of TFP or CHP against calmodulin-dependent enzymes such as phosphodiesterase is 100-400-fold higher compared to PMZ, it is speculated that the inhibitory effect of TFP or CHP against cyanide-induced convulsions may be based on its strong inhibitory properties into calmodulin-dependent enzymes. On the other hand, the LD50 value of cyanide was significantly increased by i.v.t. preadministration of TFP or CHP (0.045 mumol/body). Furthermore, the LD50 value of cyanide was also significantly increased by the same dose of PMZ. These results suggest that there is no positive correlation between mortality and convulsions induced by cyanide. PMID- 8427025 TI - Urinary excretion of porphyrins by smelter workers chronically exposed to arsenic dust. AB - Urinary arsenic (inorganic arsenic and methylated metabolites), uroporphyrin and coproporphyrin I and III isomers were determined in 84 smelter workers exposed to arsenic trioxide and in 22 non-exposed controls. Both 'high' and 'low' exposure groups were defined, based on individual's work area (arsenic recovery plant and maintenance) and mean urinary excretion of arsenic was compared to the control group (257 and 129 micrograms/g creatinine against 9.9 micrograms/g creatinine). Total coproporphyrin (I+III) increased in each exposure group as compared to control (63.3 and 59 micrograms/g creatinine against 27.2 micrograms/g creatinine), as a consequence of a 2-fold increase in each coproporphyrin isomer. The mean concentration of uroporphyrin in each exposure group was very similar to that of the non-exposed controls (9.5 and 8.8 micrograms/g creatinine against 10.7 micrograms/g creatinine). These results suggest that long-term occupational exposure of humans to arsenic is associated with coproporphyrinuria and raise the question of the use of this parameter in addition to urinary arsenic for biological monitoring. PMID- 8427024 TI - The effects of sevoflurane on intracellular Ca2+ regulation in rat hepatocytes. AB - The effects of sevoflurane, a new volatile anesthetic agent undergoing clinical trial, on the mobilization of intracellular Ca2+ in isolated rat hepatocytes was studied. This agent produced a dose-dependent release of 45Ca2+ from internal, non-mitochondrial stores of permeabilized hepatocytes (saponin treated). However, the administration of sevoflurane to aequorin-loaded intact hepatocytes had little or no effect on intracellular [Ca2+] (i.e., short transient or no increases in luminescence: no toxic effect). These data may indicate that because of the low solubility of sevoflurane, it has a selective effect on endoplasmic reticulum, i.e., mobilizing internal stores of Ca2+ relative to increasing transmembrane fluxes. PMID- 8427026 TI - Neuroendocrine granules. PMID- 8427027 TI - Argyrophilic neuroendocrine carcinoma of the male breast. AB - A case of breast carcinoma with neuroendocrine features occurring in an elderly male patient is reported. Histologically, the tumor was characterized by solid growth with nests and ribbons of small to medium-size uniform neoplastic cells. Argyrophilia, expression of chromogranins at both protein and gene level, and the presence of dense granules of the neurosecretory type by electron microscopy were demonstrated. PMID- 8427028 TI - Fibro-osseous lesion with calcified spherules (cementifying fibromalike lesion) of the tibia. AB - The histologic and ultrastructural features of a tibial lesion strongly resembling cementifying fibroma of the jaws are described. Histologically the lesion consisted of fibrous tissue containing small calcified spherules. In some areas acellular trabeculae or islets with lobulated margins of a cementlike substance occurred. Spherules and small calcified bodies were composed of apatite crystal depositions along the collagen fibers and were often in close contact with membrane-surrounded cytoplasmic processes resembling matrix vesicles. In fibrous tissue fibroblasts, myofibroblasts, and undifferentiated cells with numerous cytoplasmic processes and intracytoplasmic microfibrils were present. The character of the cellular component of this lesion and the presence of calcified spherules arranged in a manner rarely observed in fibrous dysplasia indicate a histogenic relationship between the two lesions. PMID- 8427029 TI - Use of the uranaffin reaction in the identification of neuroendocrine granules. AB - This overview emphasizes the utility of the uranaffin reaction in the diagnosis of tumors derived from neuroendocrine cells. The history, cell organelle specificity, tissue specificity, pH requirements, and detailed procedure of the uranaffin reaction is provided. Uranaffin-positive granules are also identified within the NS granules of the stem cell paraneuron (archiparaneuron) of coelenterates, and a hypothetical evolutionary scheme depicting the possible origins of the key biochemical features of the advanced mammalian NS granule is included. The role of nucleotides, a major component of true NS granules, is discussed. A possible intragranular function of ATP as a regulator of osmotic pressure and the extracellular physiologic effects of secreted intragranular nucleotides are discussed in some detail. PMID- 8427030 TI - Immunogold detection of chromogranin A in the neuroendocrine tumor. AB - Immunogold ultrastructural localization of chromogranin A in secretory granules within tumor cells provides convincing evidence of endocrine or neuroendocrine differentiation. A modified immunogold method (designed for use on osmicated tissue) produced positive labeling of small granules not only in well differentiated tumors but also in poorly differentiated small cell tumors primary in lung, cervix, and skin; only a proportion of granules in some of the tumor cells were positively labeled. Many non-small cell lung tumors often stain focally positive for chromogranin A at the light microscopy level, and such tumors may also contain sparse, small, dense granules. Because positive labeling could not be demonstrated over small granules in non-small cell lung tumors, the theory that such tumors are neuroendocrine in type may be erroneous. PMID- 8427031 TI - Ultrastructural immunolabeling in the evaluation, diagnosis, and characterization of neuroendocrine neoplasms. AB - Neuroendocrine neoplasia represents a heterogenous entity with variable morphologic light microscopic expressions. In many cases a definite diagnosis is easily made by light microscopic examination, but in some cases this does not suffice. In the latter instances, immunocytochemistry, ultrastructural examination, or both are required to diagnose a neuroendocrine neoplasm. However, basing a diagnosis of neuroendocrine neoplasia exclusively on the results obtained from immunocytochemical or ultrastructural evaluation of these tumors may not be entirely accurate in some instances. Ultrastructural immunolabeling plays a key role in accurately defining localization of immunoreactive substances in well-characterized neuroendocrine neoplasms, can assess colocalization of antigenic epitopes, helps define specificity and significance of immunocytochemistry results obtained at the light microscopic level, and is more sensitive than light microscopic immunocyto-chemistry. Some evolving diagnostic entities can be further characterized by utilization of ultrastructural labeling techniques. Controversies concerning the neuroendocrine nature of electron-dense structures identifiable at the ultrastructural level can be readily and accurately resolved. By providing a way to evaluate combined immunomorphologic parameters, ultrastructural immunogold labeling can settle important questions pertaining to neuroendocrine neoplasia. The present article illustrates a series of cases where the above-mentioned applications were tested. PMID- 8427032 TI - Lipopolysaccharide-independent radioimmunoprecipitation and identification of structural and in vivo induced immunogenic surface proteins of Salmonella typhi in typhoid fever. AB - The humoral response to Salmonella typhi is important for protective immunity against typhoid fever, as indicated by the protection obtained with killed cell vaccines and component vaccines (outer membrane proteins, Vi antigen) in animals and human beings. Nonetheless, analysis and interpretation of host humoral immune response to S. typhi surface antigens have been difficult because of the complex structure of the S. typhi envelope and the lack of purified reagents for detection of immune response to individual surface components. Normal and convalescent human sera from typhoid fever patients were absorbed with S. typhi lipopolysaccharide. These sera were used in radioimmunoprecipitation assays of whole S. typhi cells and S. typhi membranes labelled with either 125I or 35S methionine. This strategy has permitted the unequivocal identification of a humoral immune response to structural and in vivo induced outer membrane proteins of S. typhi. In this manner, we have identified the porins, lipoprotein, the iron starvation-induced proteins, and three proteins of 30, 18.5 and 15 kDa as surface exposed immunogens of S. typhi in patients with typhoid fever. These studies suggest that further experimental work is needed to characterize the relevance of both anti-S. typhi outer membrane protein and antilipopolysaccharide antibodies in recovery from S. typhi infections and protective immunity. PMID- 8427033 TI - Protective capacity of a temperature-sensitive mutant of Salmonella enteritidis after oral and intragastric inoculation in a murine model. AB - Temperature-sensitive (ts) mutant E/1/3 of Salmonella enteritidis was selected to evaluate its capacity to induce protective responses after peroral (p.o.) or intragastric (i.g.) inoculation to mice. This ts mutant of coasting phenotype was detected in Peyer's patches until day 4, and in spleen by days 3 and 4 after the mice were inoculated by the p.o. route with 10(10) colony forming units. Peroral immunization induced significant protection from oral challenge with 240 LD50 of the wild-type (wt) strain. Higher protection was achieved when the animals were boosted intraperitoneally after p.o. immunization. Intragastric inoculation with the same dose of the ts mutant increased both the level of protection, and colonization and persistence of the micro-organism in Peyer's patches and spleen. Immunization with a single i.g. inoculation induced 70% protection from p.o. challenge of the animals with the wt S. enteritidis. Two i.g. immunizations with E/1/3 raised the level of protection to 90%. Specific IgG, IgM and IgA antibodies, measured in plasma using a micro-ELISA method, were detected after i.g. immunization with ts mutant E/1/3. In addition, specific antibody-secreting cells were detected by means of an ELISPOT assay in spleen and mesenteric nodes of mice immunized with the ts mutant. PMID- 8427034 TI - Antibody response after influenza immunization with various vaccine doses: a double-blind, placebo-controlled, multi-centre, dose-response study in elderly nursing-home residents and young volunteers. AB - The dose effect (0, 10, 20 and 60 micrograms) of influenza subunit vaccine on the antibody response was investigated in nursing-home residents and young controls. The vaccine antigens were: A/Taiwan/1/86 (H1N1), A/Sichuan/2/87 (H3N2) and B/Beijing/1/87. For the influenza B antigen, the post-GMT and the 'percentage protective titre' increased significantly both in the young controls and nursing home residents. No dose effect was observed for the A/Taiwan, and a minor dose effect for A/Sichuan. All vaccine doses were well tolerated by both groups. We conclude from our data that higher vaccine doses may result in a better antibody response against some antigens but not against others. Therefore, in general, increasing the vaccine dose is no adequate method to improve the antibody response. PMID- 8427035 TI - Human macrophage responses to vaccine strains of influenza virus: synthesis of viral proteins, interleukin-1 beta, interleukin-6, tumour necrosis factor-alpha and interleukin-1 inhibitor. AB - Interactions between influenza viruses and human macrophages were examined to detect potential mechanisms for enhanced febrile reactions previously associated with administration of an avian-human H1N1 reassortant vaccine. Cells exposed to that strain were compared with cells exposed to wild-type and cold-adapted H1H1 and H3H2 strains and an avian-human H3N2 strain. Cells exposed to the avian-human H1N1 virus showed increased synthesis of viral neuraminidase, previously reported to induce fever-producing cytokines, but no detectable increase in production of interleukin-1 beta, interleukin-6 and tumour necrosis factor-alpha measured by immunoassay, or decrease in interleukin-1 inhibitor activity by bioassay. PMID- 8427036 TI - Host range selection of vaccinia recombinants containing insertions of foreign genes into non-coding sequences. AB - A simple yet powerful selection system was developed for the insertion of foreign genes in vaccinia virus. The selection system utilizes the vaccinia virus K1L (29K) host range gene which is located in HindIII M. This gene is necessary for growth in RK-13 cells but not in BSC40 or CV-1 cells. A vaccinia mutant (vAbT33) unable to grow on RK-13 cells was constructed having sequences at the 3' end of the K1L gene and the adjacent M2L gene deleted and replaced with the beta galactosidase gene regulated by the BamHI F (F7L) promoter. A recombination plasmid containing the hepatitis B surface (HBs) antigen gene regulated by the M2L promoter and the complete sequence of the K1L gene was used to insert the HBs gene into vAbT33. The M2L negative K1L positive recombinant was easily isolated in two rounds of plaque purification by plating the virus on RK-13 cell monolayers. The K1L gene selection system allows the isolation of recombinants arising at frequencies as low as 1/100,000. It was noted that recombinants containing vaccinia sequence duplications (promoters) resulted in intragenomic recombinations that eliminated all sequences between the duplications. A second recombination plasmid was constructed that allowed insertion into the vaccinia genome without the loss of vaccinia coding sequences. This was achieved by insertion of the pseudorabies virus GIII gene regulated by the vaccinia H5R (40K) promoter between the translation and transcription stop signals at the 3' end of the K1L gene. The K1L gene transcription stop signal thus became the stop signal for the inserted GIII gene and an upstream transcription stop signal present in the H5R promoter fragment provided the stop signal for the K1L gene. This manipulation of the vaccinia genome had no effect on the accumulation or 5' end of the M2L gene transcripts. Although the insertion lengthened the 3' end and lowered the accumulation of K1L transcripts it altered neither the virulence nor the immunogenicity of the recombinant. PMID- 8427037 TI - Murine model for evaluation of protective immunity to influenza virus. AB - We have characterized a murine model as an inexpensive, readily available and sensitive animal model for the evaluation of protective immune responses induced by various routes of administration of influenza A vaccine preparations. Using a non-mouse-adapted human influenza virus to infect unanaesthetized animals intranasally, we established that the optimum dose for infection of Balb/c mice was 10(4) plaque forming units of virus and that the optimum sampling time for measurement of virus yields in the organs of the respiratory tract was 72 h after challenge. We found that the infection was initiated in the nose and progressed by descending into the trachea and lungs over a period of days. Evaluation of protection against infection clearly showed that the tissues of the mouse respiratory tract were completely protected after administration of whole killed virus intranasally and partly protected when virus was administered subcutaneously. The protection correlated with the level of virus-specific IgA antibodies in saliva. PMID- 8427038 TI - Evaluation of vaccines and of antigen therapy in a mouse model for Brucella ovis. AB - A mouse model was developed to study Brucella ovis infection. The evolution of the number of B. ovis per spleen of mice inoculated intravenously, intraperitoneally or subcutaneously was found to be independent of the sex of the mice. The number of B. ovis increased in the spleen when increasing the challenge dose up to 1.7 x 10(7). At higher doses of challenge, the response remained constant. In this model it was observed that the inoculation of Brucella melitensis Rev 1 vaccine or subcellular B. ovis hot saline antigens during both the acute and chronic phases did not modify the time course of B. ovis infection. Finally, the model was found suitable to determine the efficacy of anti-B. ovis vaccines. B. melitensis Rev 1 (2.2 x 10(5) c.f.u.) and B. suis strain 2 (1.2 x 10(7) c.f.u.) live vaccines but not the inactivated B. melitensis H38 vaccine conferred protection against B. ovis. PMID- 8427039 TI - Immune response of chimpanzees after immunization with the inactivated whole immunodeficiency virus (HIV-1), three different adjuvants and challenge. AB - Purified whole virus preparations of HIV-1 were produced from supernatants of infected cells and concentrated 5000-fold. After inactivation with formaldehyde, the concentrates were combined with one of three different adjuvants, and used to immunize three groups of three chimpanzees each. The chimpanzees were monitored for HIV-specific humoral and cellular immune responses by ELISA, immunoblot, virus neutralization, delayed-type hypersensitivity, lymphocyte proliferation and antibody-dependent cell-mediated cytotoxicity. Weak and inconsistent responses were observed in animals that received HIV-1 formulated with alum as adjuvant, whereas HIV-1 formulated with incomplete Freund's adjuvant or an experimental adjuvant (BWZL) induced good humoral and cellular immune responses to the virus. The three animals that received HIV-1 with the BWZL adjuvant generated overall the best immune responses; therefore, 2 weeks after the sixth immunization these animals were challenged with infectious HIV-1. Despite the presence of good humoral and cell-mediated immunity, all three immunized animals and a control animal became infected within 4 weeks, as evidenced by repeated isolation of HIV 1 from peripheral blood mononuclear cells and anamnestic antibody responses. The new experimental adjuvant has to be further investigated in other vaccine trials and different animal models. PMID- 8427040 TI - Beta-propiolactone treatment impairs the biological activity of residual DNA from BHK-21 cells infected with rabies virus. AB - The effects of beta-propiolactone (BPL), an alkylating and virus inactivating agent, on the structural and in vitro biological properties of different DNA preparations from BHK-21 cells were investigated. Both uninfected and rabies virus-infected cells were used. Purified cellular DNA (celDNA) was used as the reference, and supernatants from infected cells were treated with BPL. For structural and biological studies three types of DNA preparation were tested: celDNA; purified DNA from cell (infected or uninfected) supernatant (pcsDNA) with or without BPL treatment; and residual cell DNA present in purified rabies virus (inactivated or not) preparations. Rabies infection and BPL (diluted 1:4000) treatment induced modifications in the structure of the three DNA types, including strand breaks and nicks. The damage to the DNA structure by BPL modifies the biological properties of the pcsDNA appraised by its ability to serve as the template in vitro for different polymerases. When rabies virus was inactivated with BPL diluted 1:1000 the DNA damage increased dramatically: small double-stranded DNA fragments (50-200 base pairs) were generated which could not function as templates for polymerases. PMID- 8427041 TI - World Conference on Poliomyelitis and Measles: Vaccines and Immunization. 7-12 January 1992, New Delhi, India. PMID- 8427042 TI - Heritable and life-style determinants of bone mineral density. AB - Familial resemblance in bone mineral density at five skeletal sites was measured among 160 adult members of 40 families. Each family included a postmenopausal mother, one premenopausal daughter, one son, and the children's father. Similarities in selected life-style factors thought to influence bone density, such as physical activity, smoking, alcohol use, and diet, were also evaluated. Bone density was measured by dual-energy (total body, femoral neck, and lumbar spine) or single-photon (radius and os calcis) absorptiometry. Correlation coefficients between the midparent Z score and offspring Z scores of bone mineral density ranged from 0.22 to 0.52 among daughters and from 0.27 to 0.58 among sons. Adjustment of bone density for age, height, weight, and significant life style or environmental factors yielded heritability estimates for the five skeletal sites between 0.46 and 0.62. That is, 46-62% of variance in bone density was attributable to heredity. Most estimates derived from the group of daughters were similar to those from the sons. These observations provide support for a significant contribution of heredity to bone density. However, an individual's life-style may account for a potentially large proportion of the nonheritable variance in bone density. PMID- 8427043 TI - Changes in axial bone density with age: a twin study. AB - Bone mineral density in adult life, which is an important determinant of fracture risk, is determined by peak adult bone density, achieved in early adulthood and subsequent rates of change during adult life. Cross-sectional twin and family studies indicate that the majority of population variation in bone density may be explained by genetic factors. Although there is evidence for a genetic effect on peak bone mass, it is unknown whether there is a genetic effect on rates of changes in bone density with age. Changes in lumbar spine and femoral neck bone density determined by dual-photon absorptiometry (Lunar DP3) were examined in a cohort of monozygotic (MZ, n = 21, 3 male and 18 female pairs, median age, range, 46; 24-75 years) and dizygotic twins (DZ, n = 19; 43, 25-65 years). The median follow-up was 3 years (range 1.1-5.5 years), with each subject having at least two and up to four bone density assessments. In these twins, genetic factors determine variation in rates of change (% change/year) in lumbar spine bone density, rMZ = 0.93 and rDZ = 0.51, p < 0.02 (one tailed), and Ward's triangle, rMZ = 0.60, rDZ = 0.11, p < 0.05 (one tailed). Model-fitting analysis was also consistent with a genetic effect on rates of change in bone density at the trochanteric site, although such an effect was not shown at the femoral neck. These data demonstrate, for the first time, the possible existence of genetic determinants of rates of change in bone mineral density in adults. PMID- 8427044 TI - Detection of mRNA for carbonic anhydrase II in human osteoclast-like cells by in situ hybridization. AB - Carbonic anhydrase II (CA II) plays an important role during osteoclastic bone resorption. Biochemical investigations of gene expression of CA II, however, have been hampered by difficulty in obtaining sufficient numbers of purified osteoclasts. In this study, we describe a nonradioactive, digoxigenin-labeled cDNA in situ hybridization technique capable of determining the pattern of CA II gene expression in human osteoclast-like cells (OC-like cells) at the single-cell level. The results showed that CA II mRNA was located in the cytoplasm of both imprinted and cultured OC-like cells from a giant cell tumor of bone. On the other hand, no evidence of CA II mRNA was found in either the mononuclear cells (tumor cells) of giant cell tumor of bone or osteosarcoma cells. There is a significant correlation between in situ hybridization and northern blot analysis for CA II mRNA in both the giant cell tumor of bone and the osteosarcoma. Our results also indicated that quantitation of in situ hybridization can be achieved by computed cytophotometry. PMID- 8427045 TI - Proton channel part of vacuolar H(+)-ATPase and carbonic anhydrase II expression is stimulated in resorbing osteoclasts. AB - Immobilization causes a transient increase in bone resorption and a prolonged depression of bone formation. We have studied the effect of immobilization on the expression of two proteins believed to have a major functional role in osteoclasts, the proteolipid subunit of vacuolar H(+)-ATPase (VPL) and carbonic anhydrase II (CA II). Trabecular bone from immobilized rat tibiae was analyzed using northern and slot blotting, polymerase chain reaction (PCR), and morphometric analysis. CA II and VPL transcription was rapidly stimulated in trabecular bone of immobilized rat tibiae. Osteoclast number increased and the trabecular bone volume decreased during immobilization. Fluorescein-labeled cDNA probes and a confocal laser scanning microscope were used to study the localization of VPL and CA II mRNAs in situ in osteoclasts and other bone-derived cells attached to bovine bone slices in vitro. CA II and VPL mRNA were highly expressed in actively resorbing osteoclasts, but in nonresorbing osteoclasts mRNA expression was very low or not detectable at all. These results strongly suggest that both CA II and VPL have an important functional role in bone resorption. Finally, in the bone cell population isolated for these studies, CA II was found to be highly specific for osteoclasts whereas VPL was also detected in other cell types. PMID- 8427046 TI - Fluoride-stimulated [3H]thymidine uptake in a human osteoblastic osteosarcoma cell line is dependent on transforming growth factor beta. AB - Controversy exists regarding the effect of fluoride on human osteoblast proliferation. To learn more of the cellular action of fluoride, we chose the clonal osteoblast cell line HOS TE85 as a model system. In these phenotypically osteoblast-like cells, sodium fluoride stimulated [3H]thymidine incorporation in a dose-dependent manner over the concentration range 1 x 10(-5)-2 x 10(-4) M. The fluoride-induced stimulation of [3H]thymidine uptake was dependent on cell density, being optimal at subconfluent cell numbers. Stimulation of [3H]thymidine uptake was inhibited by anti-transforming growth factor beta but not by antibody to insulin-like growth factor I or beta 2-microglobulin. Transforming growth factor beta was shown to be a biphasic stimulator of [3H]thymidine uptake in HOS TE85, with maximal stimulation occurring at 0.5 nM transforming growth factor beta. In the presence of fluoride the cells were more sensitive to stimulation by this growth factor, with maximum effect occurring at 0.1 nM. Fluoride did not increase mRNA for transforming growth factor beta following either 8 or 24 h of exposure. We conclude that fluoride activates osteoblast proliferation by modulating the cellular sensitivity to transforming growth factor beta, a known stimulator of bone growth. PMID- 8427047 TI - In vitro exposure to sodium fluoride does not modify activity or proliferation of human osteoblastic cells in primary cultures. AB - The anabolic effects of sodium fluoride (NaF) on trabecular bone mass in osteoporosis is now well established. In vivo histologic studies performed in humans and other animals have shown that fluoride induces an increase in osteoblast number at the tissue level. To determine the mechanisms of action of fluoride on osteoblasts, we studied the effects of NaF on short- and long-term cultures of human osteoblastic cells derived from bone explants obtained from 21 donors. In short-term experiments, bone-derived cells were exposed to NaF for 4 days. At doses ranging from 10(-11) to 10(-5) M, NaF did not modify the alkaline phosphatase (AP) activity or osteocalcin secretion. In long-term experiments, half the bone samples from 15 donors were cultured for 4 months in the presence of 10(-5) M NaF and the other half were maintained in NaF-free medium. Observations by light and electron microscopy disclosed no morphologic modification in bone explants after 4 months of exposure to NaF, despite an increase in the bone fluoride content. After the first month of culture, slight but not significant increases were noted in 6 of 10 cases for AP activity, 4 of 10 for osteocalcin secretion, and 5 of 7 for [3H]thymidine incorporation. After 4 months of culture in the presence of NaF, no change in AP activity or cell proliferation was noted. In contrast, the osteocalcin secretion significantly decreased (p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427048 TI - Transient recruitment of osteoclasts and expression of their function in osteopetrotic (op/op) mice by a single injection of macrophage colony-stimulating factor. AB - Severe deficiency of osteoclasts in op/op mice, caused by the absence of functional macrophage colony-stimulating factor (M-CSF), is cured by daily injections of purified recombinant human M-CSF (rhM-CSF). In this study, we found that a single injection of 5 micrograms rhM-CSF is enough for recruitment of osteoclasts in mutant mice. Osteoclast number increased during the period between 2 and 4 days after the single rhM-CSF injection. When YM175, a new derivative of bisphosphonate, was administered to the mice 4 days after rhM-CSF injection or later, osteoclasts disappeared by 3 days after YM175 administration. However, a significant number of osteoclasts were detected even at 3 days after YM175 administration when YM175 was administered 3 days after rhM-CSF injection or earlier. These results indicate that YM175 is cytotoxic only to functioning osteoclasts and that recruitment of osteoclasts is finished 4 days after a single rhM-CSF injection. The osteoclasts actively resorbed bone trabeculae for a prolonged period, demonstrating that M-CSF is not requisite for the functioning of mature osteoclasts. PMID- 8427049 TI - Cytokine production and surface antigen expression by peripheral blood mononuclear cells in postmenopausal osteoporosis. AB - It was reported earlier that IL-1 production by cultured monocytes and the ratio of helper (CD4) to suppressor (CD8) lymphocytes in peripheral blood are different in osteoporotic compared to nonosteoporotic subjects. We examined these and several other parameters related to the biosynthetic activity and differentiation status of peripheral blood mononuclear cells (PBMC) in untreated osteoporotic postmenopausal women (age 65 +/- 7, n = 46), nonosteoporotic postmenopausal women (age 55 +/- 3, n = 20), and nonosteoporotic premenopausal women (age 37 +/- 7, n = 8), as defined by spine density. We found that unstimulated monocytes from osteoporotics did not produce detectable IL-1 beta as determined by ELISA. In addition, there were no significant differences between osteoporotics and nonosteoporotics in IL-1 beta or IFN-gamma production by PBMC stimulated with OKT3, a monoclonal antibody to the T cell-receptor complex. The proliferative response of lymphocytes to OKT3 was significantly less (p < 0.02) in osteoporotics compared to nonosteoporotic post- and premenopausal women; multiple regression analysis, however, indicated that this difference was not due to bone density but to age. Flow cytometric analysis of PBMC revealed no difference between osteoporotics and nonosteoporotics in the distribution of 18 phenotypic subsets determined, including CD4- or CD8-positive lymphocytes or the ratio of CD4 to CD8 cells. Further, there was no correlation of the surface markers with bone density, the exceptions being the subsets expressing the CD3/CD56 and CD8/CD56 markers, which were inversely related to spine density in the osteoporotic women.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427050 TI - Immunolocalization of noncollagenous bone matrix proteins in lumbar vertebrae from intact and surgically menopausal cynomolgus monkeys. AB - The noncollagenous matrix proteins, composing about 10% of the organic matrix of bone, are considered important for cell matrix organization and regulation of mineralization in bone. In the present study, seven of the major noncollagenous bone matrix proteins were localized immunohistochemically in serial sections of lumbar vertebrae from 24 (12 intact and 12 ovariectomized) adult female cynomolgus monkeys (Macaca fascicularis). Osteocalcin was the only protein restricted to bone cells and mineralized bone matrix. Bone sialoprotein was present in both bone and calcified cartilage, and all the other proteins were distributed in soft tissues as well as bone. Staining for both osteocalcin and bone sialoprotein was present diffusely throughout the bone matrix, but osteonectin, osteopontin, matrix gla protein, decorin, and biglycan staining was concentrated along bone surfaces. Osteoid was negative for osteocalcin and bone sialoprotein, but all other proteins had areas of positive immunostaining within osteoid. All proteins except biglycan exhibited strong immunostaining of a subset of active osteoblasts, suggesting that they may be markers of osteoblast maturity or state of activation. The pattern of immunostaining in intact and surgically menopausal monkeys was similar, except that staining for matrix proteins concentrated along bone surfaces appeared to be more widely distributed in the surgically menopausal monkeys, probably due to the higher rate of bone formation in these animals. PMID- 8427051 TI - Pseudohypoparathyroidism with osteitis fibrosa cystica: direct demonstration of skeletal responsiveness to parathyroid hormone in cells cultured from bone. AB - A young girl had tibial osteotomies at age 14 for genu valgum and then had recurrent tibial cysts over a number of years. Hypocalcemia and hyperphosphatemia were first noted at age 21. The diagnosis of pseudohypoparathyroidism was made at age 28, when elevated plasma PTH was detected. Clinical and biochemical features, including a PTH response test and assay of RBC Gs, established the diagnosis of pseudohypoparathyroidism type 1b. Failure to suppress plasma PTH with vitamin D therapy led to an exacerbation of her cystic bone disease; there were widespread lytic lesions radiologically, most of which took up [99mTc]diphosphonate on bone scan. Microradioscopy revealed evidence of resorption of phalangeal tufts. Bone biopsy showed osteitis fibrosa cystica. During an orthopedic procedure, trabecular bone fragments were taken from her right humerus, and bone-derived cells cultured using an explant technique. The cultured cells were osteoblast like in morphology, fully responsive to PTH, cholera toxin, forskolin, and PGE1 in vitro, and had an alkaline phosphatase and osteocalcin response to 1,25 dihydroxyvitamin D3 [1,25-(OH)2D3]. Following this examination of skeletal responsiveness, attempts were made to suppress the elevated plasma PTH levels and symptomatic bone disease by optimizing therapy with oral 1,25-(OH)2D3. When bone pain associated with the cystic bone disease failed to resolve, the patient underwent total parathyroidectomy, following which the bone pain gradually resolved. This is the first direct demonstration of PTH responsiveness in cultured bone cells in the syndrome of pseudohypoparathyroidism with osteitis fibrosa cystica. PMID- 8427052 TI - Physicochemical effects of acidosis on bone calcium flux and surface ion composition. AB - Net calcium flux (JCa) from bone in vitro is pH dependent. When pH falls below 7.40, through a reduction in [HCO3-], there is both physicochemical and cell mediated JCa. To characterize the physicochemical effect of acidosis on bone we inhibited the bone-resorbing cells (osteoclasts) with the specific inhibitor calcitonin and studied the effect of acidosis on JCa and bone ion composition using an analytic high-resolution scanning ion microprobe. Neonatal mouse calvariae were cultured for 48 h in physiologically neutral pH medium (Ntl, pH = 7.41, [HCO3-] = 25 nM) or in medium that modeled metabolic acidosis (Met, pH = 7.10, [HCO3-] = 12), each with or without calcitonin (CT, 3 x 10(-9) M). There was net calcium efflux in Ntl (JCa = 631 +/- 36 nmol per bone per 48 h), which increased in Met (1019 +/- 53, p < 0.01); CT inhibited JCa in Ntl (-54 +/- 11, p < 0.01 versus Ntl), which increased in Met (197 +/- 15, p < 0.01 versus Ntl + CT). In the presence of CT the increase in JCa in Met versus Ntl represents physiochemical bone dissolution. The Ntl bone surface (approximately 2 nm in depth) was rich in Na compared to Ca (Na/Ca = 11.9, count/s of detected secondary ions), which fell in Met (Na/Ca = 6.0, p < 0.05); CT caused a further reduction of Na/Ca (3.1, p < 0.01 versus Ntl and versus Met), which was not altered in Met (2.6, p < 0.05 versus Ntl + CT).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427053 TI - Type IV collagen in normal and diseased glomerular basement membrane. PMID- 8427054 TI - Pediatric nephrology in honor of Dr. Renee Habib. PMID- 8427055 TI - Primary IgA nephropathies in children: prognosis and treatment. PMID- 8427056 TI - Hemolytic-uremic syndrome in the child. PMID- 8427058 TI - Early failures of kidney transplantation: a study of 70 cases from 801 consecutive grafts performed in children and adolescents. PMID- 8427057 TI - Genetic, clinical, and morphologic heterogeneity in Alport's syndrome. PMID- 8427059 TI - Antineutrophil cytoplasm antibodies: diversity and clinical applications. PMID- 8427060 TI - Renal lesions associated with human immunodeficiency virus infection: North American vs. European experience. PMID- 8427061 TI - Angiotensin antagonists. PMID- 8427062 TI - Resistance to diuretics: mechanisms and clinical implications. PMID- 8427063 TI - Glomerular sclerosis in transgenic mice: the Mpv-17 gene and its human homologue. PMID- 8427064 TI - Pancreatic islet transplantation in the human. PMID- 8427065 TI - Intrathymic transplantation as a model for tolerance. PMID- 8427066 TI - Immediate hypersensitivity reactions and hemodialysis. PMID- 8427067 TI - Interactions between plasma proteins and hemodialysis membranes. PMID- 8427068 TI - Lupus and lupus nephritis in children. PMID- 8427069 TI - The lessons of history. PMID- 8427070 TI - Primary malignant rhabdoid tumor of the brain: clinical, imaging, and pathologic findings. AB - PURPOSE: To describe the imaging and pathologic findings of malignant rhabdoid tumor (MRT), a rare primary brain neoplasm affecting children. METHODS: The CT and/or MR features, pathologic findings, and clinical records of three children with primary MRT of the brain were retrospectively reviewed. RESULTS: The tumors, large, left-sided cerebral masses, were intraventricular in two cases. MR images in one patient showed T1- and T2-weighted signal intensity isointense with gray matter. Multiple necrotic/cystic foci were present in all cases, with two showing a patchy pattern of enhancement on CT and MR. The diagnosis of MRT was documented by ultrastructural and immunohistochemical studies. All patients had normal abdominal CT scans, excluding the possibility of primary renal rhabdoid tumor metastatic to the brain. The disease progressed rapidly in each case, despite surgery, chemotherapy, and craniospinal irradiation, with serial imaging evidence of tumor regrowth at the primary site and the development of metastatic satellite lesions. CONCLUSIONS: The diagnosis of primary MRT of the brain can be made only pathologically; however, the nonspecific imaging findings in these cases suggest that MRT should be considered in the differential diagnosis of large childhood intracranial neoplasms. PMID- 8427071 TI - Solitary choroid plexus lipomas: CT and MR appearance. AB - Using MR imaging, the authors detected 23 cases of intracranial lipomas, among which were three cases of solitary choroid plexus lipomas. All the lipomas were located in the choroid plexus at the trigone. One of the three solitary choroid plexus lipomas was very small and could not be detected using CT. The authors believe that solitary choroid plexus lipomas are not as rare as was previously thought. PMID- 8427072 TI - Disappearance of cerebral calcification as a sign of tumor growth. AB - Calcification occurs in many benign and malignant neurologic disorders. We describe two patients with cerebral glioma, in whom the disappearance of cerebral calcification was evidence of local malignant change. We discuss the underlying chemical mechanisms that result in tissue calcification, and postulate that calcification may disappear in the presence of a malignant tumor because of a decrease in the pH of the microenvironment. PMID- 8427073 TI - Comparison of full- and half-dose gadolinium-DTPA: MR imaging of the normal sella. AB - PURPOSE: This study compares sellar intensities in 17 patients without sellar pathology using half (0.05 mmol/kg, nine patients) and full-dose (0.1 mmol/kg, eight patients) gadopentetate dimeglumine (Gd-DTPA). METHODS: Sellar MR studies of eight patients who received full and nine patients who received half-dose Gd DTPA were compared, retrospectively. Sequences included pre-, immediate, and delayed postcontrast T1-weighted coronal images (1.5 T). Intensity measurements were obtained by two observers using a uniform region of interest. RESULTS: Comparison of normalized intensities revealed no significant difference between intensities obtained from immediate half- and full-dose techniques for any of the tissues examined (Student's t test, P < .90). Delayed scans likewise demonstrated no significant intensity differences between full- and half-dose studies. CONCLUSION: Our findings suggest that a 50% reduction in dosage of Gd-DTPA for sellar MR at 1.5 T results in no significant diminution in intensity of enhancement of the pituitary gland or adjacent tissues. PMID- 8427074 TI - Sellar susceptibility artifacts: theory and implications. AB - PURPOSE: To investigate the prevalence and physical basis of a specific form of MR susceptibility artifact that may be seen in the pituitary gland near the junction of sellar floor and sphenoidal septum. MATERIALS AND METHODS: Coronal, T1-weighted MR images of the pituitary glands in 50 subjects without clinical evidence of pituitary or sphenoidal sinus disease were reviewed to determine the prevalence of a focal susceptibility artifact near the sellar floor. A plexiglass phantom was constructed to duplicate this artifact in vitro, the appearance of which was studied by varying the direction and intensity of the readout gradient. RESULTS: In the clinical studies, a focal artifact larger than 1 mm2 was observed in MR studies of seven (14%) of 50 subjects and was sufficiently large to mask or mimic pathology in all cases. The location of this artifact was always within the pituitary gland but closely related to the junction of the sphenoidal septum and sellar floor. The artifact was successfully reproduced in the phantom, and its magnitude was shown to be linearly related to the strength and direction of the readout gradient. An explanation for the focal nature and shape of this artifact is presented based on consideration of the boundary conditions of the Maxwell equations of electromagnetism. CONCLUSION: A focal susceptibility artifact may be seen on MR images of the pituitary gland closely related to the junction between the sellar floor and sphenoidal septum that may mimic or obscure a microadenoma. PMID- 8427075 TI - Duplication of the pituitary gland as shown by MR. AB - Duplication of the pituitary gland is a rare malformation. The authors describe a patient with features of the median cleft face syndrome (hypertelorism, V-shaped hairline, and developmental delay) who exhibited duplication of the pituitary gland and diffuse thickening of the hypothalamus (hypothalamic hamartoma) as shown by MR. The embryology of the developing pituitary gland and pathogenesis of pituitary duplication are discussed. PMID- 8427076 TI - Interuncal distance as a measure of hippocampal atrophy: normative data on axial MR imaging. AB - PURPOSE: To assess the effects of age and gender on the interuncal distance measured from MR images. METHODS: High-field strength MR images (axial) of 75 volunteers, 21-82 years old and free of neurologic disorders, were used to measure the interuncal distance. RESULTS: The interuncal distance increased with age (r = 0.66, P < .0001). The mean (SD) interuncal distance for all subjects was 21.4 (4) mm with a range of 12 to 29 mm. CONCLUSIONS: The interuncal distance in healthy subjects is unlikely to exceed 30 mm. Our data may be of clinical significance in view of a previous report stating that the interuncal distance may be enlarged in pathologic hippocampal degeneration, such as in patients with Alzheimer disease. PMID- 8427077 TI - Midsagittal MR measurements of the corpus callosum in healthy subjects and diseased patients: a prospective survey. AB - PURPOSE: To determine quantitatively a possible corpus callosum (CC) involvement in normal aging and white matter diseases. METHODS: Midsagittal size and signal of CC were recorded prospectively from 243 routine MR brain examinations. A midline internal skull surface (MISS) and subcutaneous fat signal intensity were measured to calculate CC/MISS and CC/fat ratios. Four groups of subjects were studied: 124 apparently healthy subjects, 45 patients with multiple sclerosis, 13 patients with a noncerebral cancer under chemotherapy, and 37 AIDS patients. RESULTS: Mean surface area of CC in controls was 6.36 cm2. It was significantly larger in men than in women (P < .05), but CC/MISS ratio was not. Elderly controls > 70 years and AIDS patients displayed significant CC atrophy, as well as multiple sclerosis subjects with long-standing disease or with a severe chronic progressive form. CONCLUSION: CC substance loss identification should not be based on visual inspection or on absolute area, but by means of a CC/MISS ratio. PMID- 8427078 TI - Pathophysiologic assessment of stagnating arteries after removal of arteriovenous malformations. AB - PURPOSE: To assess the hemodynamics and pathophysiology of stagnating arteries after removal of arteriovenous malformations (AVMs). SUBJECTS: 50 patients with supratentorial pial AVMs underwent pre- and postoperative angiographic studies. RESULTS: The following characteristics were found to correlate with stagnating arteries: 1) advanced patient age, 2) large AVM size, 3) markedly dilated feeders, 4) early postoperative angiograms, and 5) delayed restoration of feeding artery diameter. CONCLUSIONS: The rate of blood flow in the former feeding arteries, expressed as v x pi x r2 (v = mean velocity, r = vessel radius), suddenly decreases after removal of AVMs. When dilatation persists postoperatively in these arteries the flow velocity decreases and stagnation takes place. Delayed postoperative restoration of feeding artery diameter may be caused by a decrease of elasticity due to long-standing hemodynamic stresses, and by increased postoperative vascular resistance of these arteries. PMID- 8427079 TI - Hallervorden-Spatz disease: MR and pathologic findings. AB - PURPOSE: To compare the MR findings of eight cases with clinical diagnosis of Hallervorden-Spatz disease (HSD) with the pathologic findings of two other cases of HSD. MATERIALS AND METHODS: The eight imaged cases were studied with 0.5-T (seven cases) and/or 1.5-T (five cases) units. Six patients also had CT scans. The two other cases with proven HSD had detailed histologic evaluation. RESULTS: The 1.5-T findings showed abnormalities confined to the pallidum, which presented a diffuse low signal intensity in T2-weighted images, and an anteromedial area of high signal intensity (eye-of-the-tiger sign). In 0.5-T studies, low signal intensity was less evident and poorly detectable in spin echo, but gradient-echo images could enhance its demonstration; the area of high signal intensity was always well demonstrated. In three cases (three with 1.5 T, one with 0.5 T) a central spot of low signal intensity was seen in this area. The pathologic cases, in addition to neuroaxonal swellings and iron deposits, exhibited areas of "loose" tissue with vacuolization and lesser amounts of iron in the anteromedial part of the pallidum, in a location corresponding to the area of high signal intensity of the imaged cases. CONCLUSION: Comparison of MR findings with the pathologic studies demonstrates that the low signal intensity in T2-weighted images at 1.5 T corresponds to iron deposits in a dense tissue, and that the high signal intensity of the eye-of-the-tiger sign corresponds to an area of loose tissue with vacuolization. No correlation was found in the two pathologic cases for the central spot of low signal intensity. PMID- 8427080 TI - Resolving MR features in osmotic myelinolysis (central pontine and extrapontine myelinolysis). AB - Osmotic myelinolysis is a distinctive clinical syndrome with characteristic MR features in the central pons (central pontine myelinolysis) and in other locations (extrapontine myelinolysis). We describe the resolving MR features in an adolescent who has experienced complete neurologic recovery. Regions of involvement manifested increased T2 signal intensity. The extrapontine involvement was noted to resolve earlier with interim-increased T1-weighted signal. The mechanism for the variable appearance of increased T1 signal intensity is discussed. PMID- 8427081 TI - Hemorrhagic necrosis and vascular injury in carbon monoxide poisoning: MR demonstration. AB - MR imaging of a patient 3-years status post-carbon monoxide (CO) poisoning revealed areas of abnormal signal bilaterally in the region of the globus pallidus that had shorter T1 characteristics and longer T2 characteristics than cerebrospinal fluid, probably representing methemoglobin, and that is surrounded by a rim of decreased signal on T2-weighted images, felt to represent hemosiderin. This case demonstrates characteristic findings on MR imaging of CO poisoning, as well as observations that suggest prior focal hemorrhage. Typical findings, neuropathology, and the role of vascular injury and prognosis are discussed. PMID- 8427082 TI - Intraarterial use of sodium methohexital for provocative testing during brain embolotherapy. AB - PURPOSE AND METHODS: To assess its vascular effects and safety, we used sodium methohexital (Brevital)--an ultrashort-acting barbiturate--as the provocative intraarterial agent in a series of 30 patients with arteriovenous malformations at a 1% concentration and at doses of less than 5 mg per injection. Digital vascular imaging was performed just prior to and just after the injections. RESULTS: No angiographic or clinical evidence of apparent vasospasm occurred in the trial population (66 vascular pedicle injections in 30 patients). When functional tissue was perfused with 1-6 mg of the 1% Brevital solution, evidence of altered neurologic status became immediately apparent, but cleared within 2 minutes in all cases. None of the patients experienced either prolongation of the induced clinical symptoms or seizures to suggest any adverse effects related to either crystallization of the Brevital or the effects of injecting an alkaline solution in the cerebral circulation. CONCLUSION: Though the full effects of methohexital in the cerebral circulation remain to be elucidated, existing reports suggest it is a safe provocative agent for use prior to embolotherapy for brain arteriovenous malformations. PMID- 8427083 TI - Deflation of detachable balloons in the cavernous sinus by percutaneous puncture. AB - The authors show that percutaneous puncture of balloons within the cavernous sinus is technically feasible and allows further access to the cavernous sinus after balloon detachment. Complete closure of a large carotid cavernous fistula was achieved in the 37-year-old trauma victim they treated using this technique. PMID- 8427084 TI - Percutaneous catheter placement for cyst drainage in the subarachnoid space. AB - A technique for percutaneous catheter drainage of cystic masses in the subarachnoid space was developed and shown to be safe and effective in an 18-year old boy with life-threatening, recurrent neuroenteric cysts that compressed the brain stem, cerebellum, and spinal cord. Percutaneous drainage through a C1-C2 approach was performed on 11 separate occasions. Decompression was always accomplished and no infection or other complication occurred, even with continuous catheter drainage for 9 months. This technique provides opportunities for interventional neuroradiologic therapy in the subarachnoid space. It appears to be suited for percutaneous drainage of intraspinal and intracranial cysts when surgery is not indicated due to intractability or inaccessibility. PMID- 8427085 TI - Intracranial extension of an idiopathic orbital inflammatory pseudotumor. AB - Idiopathic orbital inflammatory pseudotumor (IOIP) with endocranial extension is very unusual. The authors used CT and MR to diagnose IOIP and demonstrate the presence of intracranial extension of orbital and lacrimal gland lesions. While providing additional evidence of IOIP having intracranial extension, this case report emphasizes the need to include IOIP as a possible differential diagnosis when radiologic explorations reveal lesions extending from the orbit to intracranial structures. PMID- 8427086 TI - Herpes zoster ophthalmicus with orbital pseudotumor syndrome complicated by optic nerve infarction and cerebral granulomatous angiitis: MR-pathologic correlation. AB - The authors describe a 41-year-old woman with herpes zoster ophthalmicus and extensive intracranial and orbital involvement as documented by MR and pathologically. MR showed all of the lesions that led to the ophthalmoplegia and pseudotumor syndrome, the periaxial infarct of the distal optic nerve, pontine infarcts, and granulomatous angiitis of the meningeal vessels. MR is useful in both detection and monitoring of the disease. PMID- 8427087 TI - Qualitative phase contrast MRA in the normal and abnormal circle of Willis. AB - PURPOSE: To determine the direction of blood flow in the circle of Willis using a 3-D phase contrast MR angiographic (MRA) technique with high spatial resolution. SUBJECTS: Fifty healthy subjects and 15 patients with occlusive disease were studied using 3-D phase contrast MRA. RESULTS: In the 50 normal subjects, 39 (78%) had detectable flow in one or both posterior communicating arteries. In 24 (48%) of these subjects, flow was detected in both posterior communicating arteries, whereas unilateral flow was detected in 15 (30%). In 36 (92%) of the 39 normal subjects, flow in the posterior communicating artery was from anterior to posterior with only 3 (8%) showing reverse flow from posterior to anterior. The A1 segment of both anterior cerebral arteries was identified in 100% of normal subjects with flow in the expected direction from carotid to the A2 segment. In patients with carotid occlusion, the pattern of flow in the circle of Willis was altered with reversed flow in the ipsilateral posterior communicating artery and sometimes in the ipsilateral A1 segment. An ipsilateral posterior communicating artery was present in 10 of 17 occluded carotid arteries, all showing reversed flow. CONCLUSION: 3-D phase contrast MRA provides useful information about the hemodynamics of normal and abnormal blood flow in the circle of Willis. PMID- 8427088 TI - Amaurosis fugax caused by a dural arteriovenous fistula from the ophthalmic artery. AB - A 52-year-old man presented with transient monocular blindness that was both spontaneous and exacerbated by exertion. Dynamic orbital CT revealed a delay in the perfusion of the left optic nerve head suggestive of a steal phenomenon. Subsequent selective arteriography demonstrated an arteriovenous fistula between the falx artery originating from the ophthalmic artery and the superior sagittal venous sinus. In the proper clinical setting, a hemodynamic steal should be considered in the differential diagnosis of amaurosis fugax. PMID- 8427089 TI - MR of enhancing nerve roots in the unoperated lumbosacral spine. AB - PURPOSE: To evaluate prospectively the unoperated lumbosacral spine by gadolinium enhanced MR for evidence of clinically correlative abnormal enhancement of nerve roots. METHODS: Two hundred patients were prospectively studied with intravenous gadolinium- (0.1 mmol/kg) enhanced MR. Evidence was sought of intrathecal enhancement of lumbosacral nerve root(s), and the correlation of this enhancement with the clinical syndrome was evaluated. RESULTS: Ten patients demonstrated abnormally enhancing lumbosacral nerve root(s) (5%). Of these, seven (70%) were associated with focally protruding disk pathology. The three (30%) remaining patients had isolated enhancement of multiple nerve roots in the absence of significant associated anatomic pathology. Overall, the correlation of radicular enhancement with the presenting clinical syndrome was excellent. CONCLUSION: A breakdown in the blood-nerve barrier as observed on gadolinium-enhanced MR serves as a marker for nerve root pathology in the unoperated lumbosacral spine, which may have clinical relevance in certain clinical situations. PMID- 8427090 TI - MR of the spinal cord in a patient with herpes zoster. AB - Increased signal intensity on initial magnetic resonance images of the spinal cord in a patient with herpes zoster demonstrated that this virus caused inflammation of the cervical spinal cord. This pathology corresponded well with neurologic deficits seen clinically, but the extent of the neurologic deficits ultimately could not be determined by magnetic resonance of the spinal cord alone because the nerve roots were also affected. PMID- 8427091 TI - Extraosseous spinal chordoma: radiographic appearance. AB - The authors describe a chordoma arising within the soft tissues of the spinal canal (rather than from a vertebral body) in a 6-year-old boy. The lesion was entirely extradural as shown by CT and MR; this was confirmed at surgery. PMID- 8427092 TI - Warthin tumor of the parotid gland: MR-pathologic correlation. AB - PURPOSE: To describe the MR appearance of Warthin tumor and correlate the images with pathologic sections. METHODS: MR studies of seven patients with Warthin tumors in the parotid gland were retrospectively reviewed; MR results were compared with pathologic specimens in five instances. FINDINGS: One patient had a bilateral tumor; another had three tumors in the right parotid gland and one in the left. Ten tumors showed well-defined borders; the other, which was accompanied by infection, had an indefinite border only at the upper pole. All had thin capsules not shown on MR. Four showed small lobulations at the margins. Fibrosis appeared as septa-like structures of low signal intensity in three cases. There were areas of hemorrhage with accumulation of hemosiderin in two cases. All tumors had low or intermediate T1-weighted signal intensity, four with high intensity areas of cysts containing cholesterol crystals. On T2W images, two tumors had homogeneously intermediate intensity; the other nine had mixed intensity. Areas of intermediate intensity were correlated with abundant epithelial tissues; focal high-intensity areas were correlated with cysts and/or areas of predominant lymphoid proliferation. CONCLUSIONS: MR findings such as bilaterality and/or multiplicity, well-defined margins, and predominantly intermediate signal intensity on T1W and T2W images with focal areas of high signal intensities on T1W images suggest Warthin tumor, but are not pathognomonic. PMID- 8427093 TI - Amyloidoma of the nasopharynx: CT and MR findings. AB - A case of nasopharyngeal amyloidoma with extensive skull base erosion is presented. CT revealed a large, relatively homogeneous, enhancing lesion; MR revealed a signal intensity iso- or slightly hyperintense compared to muscle on T1- and T2-weighted images, with a moderate degree of contrast enhancement. When an erosive mass is encountered at the skull base in a submucosal location in the nasopharynx, and MR demonstrates short T2 relaxation with signal iso- or slightly hyperintense relative to muscle, amyloidoma should be included in the differential diagnosis. PMID- 8427094 TI - Sinus histiocytosis with massive lymphadenopathy: a case of simultaneous upper respiratory tract and CNS disease without lymphadenopathy. AB - The authors describe a 20-year-old man who initially developed an intradural mass in the upper cervical region and subsequently presented with nasal/paranasal sinus and posterior fossa masses, well demonstrated by CT and MR. Histopathology demonstrated dense fibrous tissue, aggregates of histiocytes with round to oval vesicular nuclei, and other features diagnostic of Rosai-Dorfman disease; a right nasal biopsy prior to surgery showed similar microscopic findings. PMID- 8427095 TI - Ameloblastoma of the maxilla: CT and MR appearance. AB - This paper describes the CT and MR appearance of an ameloblastoma that involved the maxilla, infratemporal fossa, and adjacent structures. Although not pathognomonic, the multicystic appearance of an ameloblastoma may suggest the correct diagnosis. PMID- 8427096 TI - Superficial siderosis of the CNS: MR diagnosis and clinical findings. AB - PURPOSE: To report the clinical and neuroradiologic findings of superficial siderosis of the CNS, due to chronic subarachnoid bleeding of unknown origin. MATERIALS AND METHODS: We observed seven cases. The main clinical manifestations were progressive deafness and ataxia. Four patients had had previous cranial or cervical trauma, with root avulsion in two, many years before onset of deafness and ataxia. Neuroradiologic studies included MR (0.5 T in four and 1.5 T in three) and angiography of the brain in all cases, CT in six cases, MR of the spine in six, and myelography in four. RESULTS: MR demonstrated a rim of marked hypointensity in T2-weighted images, consistent with hemosiderin deposits, on the surface of cerebellum, brain stem, inferior part of cerebral hemispheres, and spinal cord. CT showed cerebellar atrophy in five cases, and a rim of mild hyperdensity around the brain stem in two. Angiographic studies were negative. Myelography showed cervical nerve root avulsion in two cases and a cervicodorsal extradural cyst in one. Cerebrospinal fluid contained RBCs in all the six examined cases. CONCLUSION: Although CT may occasionally suggest the diagnosis of superficial siderosis, MR demonstrates this abnormality to better advantage. PMID- 8427097 TI - Meningiomas in children: MR and histopathologic findings. AB - PURPOSE: To present the MR and histopathologic findings in five children with meningiomas. METHODS: Five children aged 3 months to 16 years with pathologically proved meningiomas underwent preoperative contrast enhanced MR. Tissue in four patients was sent for chromosomal analysis in addition to routine histopathologic studies. RESULTS: All five tumors were extra-axial, two supratentorial and three in the posterior fossa. MR showed variable signal intensity on precontrast T1 weighted images. All of the tumors were hyperintense on proton density- and T2 weighted images and showed intense contrast enhancement. Histopathologic analysis showed two meningotheliomatous, one transitional, one chordoid, and one hemangiopericytic variant of meningioma. Chromosomal analysis showed deletions involving chromosome 22 in two of four tumors studied. CONCLUSION: Meningiomas in children have a higher incidence of posterior fossa location and different histologic types than seen in adults. MR showed the tumors in our patients to be extra-axial, hyperintense on proton density- and T2-weighted images with intense enhancement on postcontrast T1-weighted images. Chromosomal aberrations were noted in two patients. PMID- 8427098 TI - Direct origin of the artery of the cervical enlargement from the left subclavian artery. AB - An anatomic variation is described in which the principal radiculomedullary artery to the cervical spinal cord, the artery of the cervical enlargement, arises directly from the left subclavian artery. This anomaly is important clinically because it may be necessary to catheterize this vessel selectively during spinal arteriography, and also because unintentional injection of this vessel can be associated with complications. PMID- 8427099 TI - Xenon/CT cerebral blood flow studies during continuous depth electrode monitoring in epilepsy patients. AB - PURPOSE: To observe and describe cerebral blood flow (CBF) alterations immediately following depth electrode stimulation of the temporal lobe in patients with medically intractable epilepsy. MATERIALS AND METHODS: Five patients with partial epilepsy undergoing presurgical evaluation were chosen for xenon/CT cerebral blood flow (Xe/CT CBF) measurement immediately following electrically stimulated seizures via stereotactically placed temporal lobe depth electrodes. Each patient had a baseline Xe/CT CBF study. Four of the five patients had a total of seven temporal lobe stimulations each followed by a Xe/CT CBF study. The other patient had right temporal lobe electrical status epilepticus and was scanned without stimulation or electroencephalogram monitoring. RESULTS: Of the four baseline or interictal scans, no areas of abnormally low flow were detected, but one baseline scan had elevated flows of 115 mL.100 g-1.min-1 in the left temporal lobe. One stimulation elicited 8 seconds of afterdischarge potentials, but no alteration of CBF was detected. One stimulation elicited an aura but no electrographic seizure was detected. This resulted, however, in bitemporal lobe elevation of CBF. The other five temporal lobe stimulations resulted in 17-63 seconds of afterdischarge potentials and all resulted in elevation of CBF to 69-118 mL.100 g-1.min-1. One of these five stimulations resulted in seizure and localized elevation of CBF. Following seizure activity, elevated CBF began to return to baseline levels by 20 minutes. CONCLUSION: This study reveals a direct spatial and temporal relationship of elevated CBF with seizures. This study provides the most direct data to date in human subjects that focal seizure activity elevates CBF. Since seizures are known to increase metabolic activity in the activated tissue, this data also supports the assumption of coupling between CBF and metabolism during the pathologic process of a seizure. PMID- 8427100 TI - Giant cranial hemangiopericytoma: MR and angiographic findings. AB - The authors detail the MR and angiographic findings of a very large hemangiopericytoma in the skull of a 63-year-old woman. Angiography showed a marked tumor blush with early draining veins; MR showed heterogeneously increased T2 signal intensity and enhancement with gadolinium secondary to the richly vascularized tumor. PMID- 8427101 TI - Efficacy of gadolinium in MR brain imaging of HIV-infected patients. AB - PURPOSE: To determine the value of gadolinium in routine head MR imaging of HIV infected patients. METHODS: One hundred and three consecutive human immunodeficiency virus-infected patients referred for head MR imaging were scanned without and with intravenous gadopentetate dimeglumine (Gd-DTPA) contrast. RESULTS: The precontrast scans of 82 patients were either normal, or had atrophy or diffuse white matter changes only. Sixteen of these 82 demonstrated enhancing abnormalities: eight meningeal/ependymal enhancement and eight focal enhancing lesions. Twenty-one of the 103 scans had focal or mass lesions on the precontrast images; in eight of these scans, new information was obtained with Gd-DTPA. Of the 24 patients in both groups where new information was obtained with Gd-DTPA, the information contributed to a change in the clinical care of nine patients. CONCLUSION: Gadolinium-enhanced MR is useful in the management of selected patients with human immunodeficiency virus infection, for example those with symptoms suggesting meningeal involvement, focal brain lesions, or if the unenhanced MR does not explain all the patient's symptoms. PMID- 8427102 TI - Vasculitis presenting as primary leptomeningeal enhancement with minimal parenchymal findings. AB - In this case of biopsy-proved granulomatous angiitis, MR showed prominent leptomeningeal enhancement with comparatively little parenchymal involvement. Pathologically, granulomatous angiitis chiefly affects small vessels, with a predilection for those in the leptomeninges. Although marked leptomeningeal enhancement is not the most common finding, vasculitis should be added to the differential diagnosis of this MR appearance. PMID- 8427103 TI - Posttraumatic pseudoaneurysm of the posterior meningeal artery associated with intraventricular hemorrhage. AB - A 22-year-old man sustained a severe head injury and had a torn posterior meningeal artery that caused massive intraventricular hemorrhage. Traumatic pseudoaneurysm of the posterior meningeal artery should be considered in cases where intraventricular hemorrhage occurs in the presence of occipital bone fracture and contiguous epidural hematoma; vertebral angiography is of value in this regard. PMID- 8427104 TI - Summary of the proceedings of the fourth annual meeting of the Eastern Neuroradiological Society. PMID- 8427105 TI - Epidural pneumatosis associated with spontaneous pneumomediastinum. PMID- 8427106 TI - Angiography of encephalomyosynangiosis and superficial temporal artery to middle cerebral artery anastomosis in moyamoya disease. AB - The authors present an angiographic follow-up of anastomotic surgery (extra- to intracranial) in a 33-year-old woman with moyamoya disease. The bypass was superficial-temporal to middle-cerebral, with placement of muscle tissue over the open craniotomy; the bypass eventually regressed. PMID- 8427107 TI - Neuroradiologic evaluation of patients with acute stroke treated with recombinant tissue plasminogen activator. The rt-PA Acute Stroke Study Group. AB - PURPOSE: 1) To describe the effectiveness and safety of thrombolytic therapy in patients with acute atherothrombotic and embolic stroke and 2) to study the variables of the occlusion site as seen on the angiograms, the CT signs of early ischemia, the hyperdense middle cerebral artery sign (HMCAS), and the size of the infarcts as seen on the 24-hour CT scan. METHODS: Ninety-three of 139 patients with acute stroke were treated with intravenous tissue plasminogen activator (rt PA). The initial disease and the effects of treatment were assessed with both CT and cerebral angiography. RESULTS: Recanalization of occluded arteries occurred in 32 patients and was more frequent in distal occlusions. In general, patients displaying recanalization tended to develop small infarcts and patients with a HMCAS tended to develop large infarcts. Patients with signs of early ischemia developed large infarcts. The presence of a HMCAS was 100% fic for an occluded artery, but only 27% sensitive. Hemorrhagic transformations occurred in the distribution of the occluded arteries in 32 patients. CONCLUSIONS: Emergency cerebral angiography, which can be carried out relatively safely, adds important information about the nature and extent of the arterial occlusions, and the recanalization efficacy of fibrinolytic therapy for patients with acute stroke. Fibrinolytic therapy can be carried out with a relatively low complication rate that still needs to be correlated with the clinical benefits of the treatment. Fibrinolytic therapy in the doses utilized in this study, is more effective with distal than with proximal carotid territory occlusions. PMID- 8427108 TI - Multiple asymptomatic cervical cephalic aneurysms. AB - The authors describe the clinical and radiologic features of pseudoaneurysms of the extracranial carotid and vertebral arteries in a 35-year-old woman, presumably resulting from trauma from previous seizures. PMID- 8427109 TI - MRA delineation of the vertebral-basilar system in patients with hemifacial spasm and trigeminal neuralgia. AB - Detailed depiction of the vertebral-basilar system is often obscured by other vascular structures on the MR angiogram. A special MR angiography technique that can better delineate the branches of the vertebral-basilar system has been designed and has proved particularly useful in the identification of tortuous vascular branches when they come in contact with the seventh or fifth cranial nerves. PMID- 8427110 TI - Sensorineural hearing loss: more than meets the eye? AB - PURPOSE: To assess the value of MR in patients with sensorineural hearing loss (SNHL) caused by lesions other than acoustic neuromas. METHODS: MR studies of 51 patients with SNHL were retrospectively reviewed; patients with acoustic neuroma were excluded to focus on the more uncommon causes. RESULTS: Twenty patients had labyrinthine lesions. Six patients had viral labyrinthitis, one patient had bacterial labyrinthitis, and one patient had luetic labyrinthitis. Three patients had hemorrhage in the labyrinth, two posttraumatic and one spontaneous from an adjacent temporal bone tumor. Only one of the two patients with traumatic labyrinthine hemorrhage had evidence of a fracture on high-resolution CT. In one patient with CT-proved cochlear otosclerosis, peri-cochlear foci of enhancement were seen on contrast-enhanced MR. Four patients had presumed labyrinthine schwannomas. A middle ear cholesteatoma in one patient invaded the cochlea and resulted in marked cochlear enhancement due to granulation tissue. Thirteen patients had intracanalicular and cerebellopontine angle lesions. The lesions included arteriovenous malformations (three patients), sarcoidosis (three patients), metastasis (two patients), lymphoma (two patients), lipomas (two patients), and postshunt meningeal fibrosis (one patient). Eighteen patients had intra-axial lesions responsible for SNHL. The most common intra-axial lesions were brain stem infarcts and multiple sclerosis. Traumatic lesions in the inferior colliculi, sarcoidosis, lymphoma, and extrinsic compression of the colliculi from a pineal tumor were also noted. CONCLUSION: MR can demonstrate numerous lesions responsible for SNHL other than acoustic neuromas. The entire acoustic pathways, including the labyrinth, internal auditory canal, cerebellopontine angle, and brain stem should be carefully scrutinized for lesions in patients with SNHL. The use of contrast media markedly increases the yield of MR in this clinical situation by demonstrating inflammatory and neoplastic labyrinthine lesions and meningeal pathology (both neoplastic and inflammatory) in the internal auditory canal and cerebellopontine angle cistern. PMID- 8427111 TI - Constructive interference in steady state-3DFT MR imaging of the inner ear and cerebellopontine angle. AB - PURPOSE: To assess the value of a three-dimensional Fourier transformation MR technique "CISS" (constructive interference in steady state) in imaging the inner ear. SUBJECTS: We studied 50 normal inner ears (40 axial, 10 coronal) and 10 pathologic inner ears in 60 patients. RESULTS: The cochlea, semicircular canals, and vestibulum were visualized in detail. Cranial nerve VII and the cochlear, superior vestibular, and inferior vestibular branch of cranial nerve VIII were identified in 90%, 94%, 80%, and 88% of the cases, respectively. A vascular loop was recognized inside the internal auditory canal in 6%, and in the porus in 30%, of the cases. The high signal of the cerebrospinal fluid and labyrinthine fluids (perilymph and endolymph) on the CISS images made excellent delineation of tumors in the cerebellopontine angle and internal canal possible and allowed detection of tumoral labyrinth involvement. The thin sections, high resolution of the images, and capability of producing multiplanar and three-dimensional reconstructions often offered additional information. CONCLUSIONS: The CISS sequence allows detailed study of the normal and pathologic inner ear and promises to be highly valuable in the demonstration of the vascular loop. PMID- 8427112 TI - Pathology of the membranous labyrinth: comparison of T1- and T2-weighted and gadolinium-enhanced spin-echo and 3DFT-CISS imaging. AB - PURPOSE: To assess the value of unenhanced T1-weighted images, T2-weighted images, gadolinium-enhanced T1-weighted images, and three-dimensional Fourier transformation-constructive interference in steady state (3DFT-CISS) images in depicting lesions of the membranous labyrinth. METHODS: Six patients were studied using 1-T MR; both enhanced (gadolinium-tetraazacyclododecane tetraacetic acid) and unenhanced images were obtained and different sequences compared to determine which provided the most information. RESULTS: A combination of gadolinium enhanced T1-weighted and 3DFT-CISS images could depict all membranous labyrinth pathology. Unenhanced T1-weighted images were necessary to exclude spontaneous hyperintensity in the membranous labyrinth. Gadolinium-enhanced T1-weighted images were needed to detect enhancing pathology such as labyrinthitis and tumors inside the membranous labyrinth. In these cases, 3DFT-CISS images allowed immediate differentiation between inflammation and tumor. In temporal bone tumors involving the bony and membranous labyrinth, unenhanced and enhanced T1-weighted images often sufficed to suggest the correct diagnosis. Only 3DFT-CISS images were able to demonstrate small structures (as fistulas) and to help us confirm or rule out obliteration of the labyrinthine fluid spaces. 3DFT-CISS images were necessary to detect small congenital malformations of the membranous labyrinth when only MR was performed. Uncalcified obliteration of the labyrinth fluid spaces could be reliably detected only on 3DFT-CISS images. Here also gadolinium enhanced T1-weighted images had to be obtained because enhancement of the soft tissues inside the membranous labyrinth had been observed. CONCLUSION: The CISS sequence and enhanced T1-weighted sequence formed the best sequence combination for diagnosis of membranous labyrinth lesions; additional, unenhanced T1-weighted images can help one differentiate labyrinthitis, proteinaceous fluid, subacute hemorrhage, or tumor inside the labyrinth. PMID- 8427113 TI - Chondroblastoma of the temporal bone: CT and MR appearance. AB - The authors report a case of chondroblastoma in the unusual location of the temporal bone. CT findings do not differ dramatically from other reports: the lesion appeared as a soft-tissue density mass in the right temporal bone, with bony destruction and thinning of cortical margins. MR findings are more rare: on coronal T1-weighted images the lesion appeared as a mass, isointense to gray matter, centered in the right petrous bone; on axial T2-weighted images, as a mixed-intensity signal mass. They conclude that MR is an accurate indicator of the location and extension of the tumor, but that CT gives more specific information regarding bone involvement. PMID- 8427114 TI - CT of the temporal bone in a patient with osteopathia striata and cranial sclerosis. AB - The authors present a case of congenital dysplasia affecting the long bones, skull, and other systems in a 7-year-old girl, with special attention to CT of the temporal bone, which showed abnormal ossicle fixation, a narrowed Eustachian canal, thickened sclerotic bone, and a small mastoid antrum and middle ear cavity. CT of the temporal bone can help one distinguish the etiologies of hearing loss associated with this disorder. PMID- 8427115 TI - Dynamic contrast enhancement of intracranial tumors with snapshot-FLASH MR imaging. AB - PURPOSE: To investigate whether exact evaluation of the dynamic contrast enhancement pattern within intracranial tumors can help to classify tumor histology and vascularity. METHOD: Forty-nine patients suffering from different intra- and extraaxial intracranial tumors underwent MR-examination in a 1.5-T superconductive whole body system. After rapid injection of Gd-DTPA, 48 images were acquired during the first 1.5 to 2 minutes of contrast enhancement within the tumors. A fast snapshot-FLASH imaging technique allowed measurement times of 1 second per image. Appearance of Gd-DTPA in a venous sinus served as a temporal reference point. Transformation of 48 discrete measurement points (mean signal values of the enhancing tumor region) into a continuous curve, using a cubic spline approximation, allowed calculation of the time of maximum signal increase (Tm1) and the following time of half maximum increase (Tm2). These time parameters were compared to histopathologic findings, especially the degree of tumor vascularization. RESULTS: Significantly different dynamic patterns of the early enhancement period were found for the different tumors. All eight neurinomas, typically less vascularized than most meningiomas, showed a characteristically prolonged contrast enhancement with a long Tm2. Histopathologic findings concerning the degree of vascularization showed two subtypes in meningiomas (n = 17) as well as in pituitary macroadenomas (n = 7). This was confirmed by dynamic evaluation in all cases, in the sense that short Tm1 and Tm2 were found in cases with higher degrees of vascularization. Negatives values of Tm1 were measured in two glomus jugulare tumors, reflecting the arterialization of these vascular tumors. In neuroepithelial tumors (n = 15), the glioblastomas (n = 7) showed very short Tm1 compared to the lower grade gliomas (n = 8). This is explained by histologic findings of pathologic vessels with arteriovenous shunts. CONCLUSION: The evaluated dynamic time parameters can be used to narrow differential diagnostic possibilities and to infer the degree of vascularization of intracranial tumors. PMID- 8427116 TI - Metastatic lesions involving the cerebellopontine angle. AB - PURPOSE: To evaluate the clinical and MR findings of metastatic lesions involving the cerebellopontine angle (CPA), which may be useful in differentiating them from the more commonly occurring benign CPA lesions. METHODS: Clinical and MR findings of 14 patients with clinical/radiologic (seven) or pathologic (seven) diagnoses of CPA metastasis were retrospectively reviewed. RESULTS: Useful clinical findings included acute onset and rapid progression of cranial nerve symptoms, especially 7th and/or 8th cranial nerve deficits (92.9%). Cranial nerve symptoms could be unilateral (50%) and frequently involved multiple cranial nerves (64.3%). MR findings showed significantly more extensive disease than suggested by clinical presentation, with 100% of patients having multiple cranial nerve involvement and 85.7% bilateral. Useful MR findings included small and/or bilateral CPA-enhancing lesions with relative isointensity to brain parenchyma on precontrast MR, with associated findings of multiple and/or bilateral cranial nerve and/or leptomeningeal lesions. CONCLUSIONS: These associated findings suggest that cerebrospinal fluid dissemination and/or leptomeningeal extension may be an important pathway for metastatic spread to the CPA. Because the CPA metastasis may be the initial or only site of metastasis, and may occur many years after the initial diagnosis of malignancy, MR findings with clinical correlation are not only useful for the detection of CPA metastases, but also for their differentiation from the more common benign CPA tumors. PMID- 8427117 TI - Increased production of thromboxane A2 by coronary arteries after thrombolysis. AB - The coronary artery produces large amounts of prostacyclin (PGI2) and a small amount of thromboxane A2 (TXA2); this high PGI2/TXA2 ratio contributes to the antithrombotic properties of the coronary artery. This study was designed to determine whether this ratio changes after coronary artery thrombosis and thrombolysis and accounts for coronary artery reocclusion. Anesthetized dogs (N = 12) were subjected to electrically induced coronary artery thrombosis and tissue plasminogen activator-induced thrombolysis. Thrombolysis was achieved in 11 dogs, and the coronary artery reoccluded in five of these dogs after the initial reperfusion. Spontaneous and ionophore A23,187-stimulated PGI2 and TXA2 synthesis in normal circumflex and ischemic-reperfused left anterior descending coronary artery segments was measured by radioimmunoassay of thromboxane B2 and 6-keto prostaglandin F1 alpha, respectively. Production of TXA2 was 413% to 656% greater in left anterior descending segments (from the region of thrombosis and sites proximal and distal to the thrombus) compared with normal circumflex segments (p < 0.001). Production of PGI2 was also increased but only by 46% to 80% in the left anterior descending segments compared with normal circumflex segments (p < 0.05). TXA2 production was greater in coronary artery segments that reocculded compared with segments that stayed open (p < 0.02). Scanning electron microscopy revealed platelet deposition in thrombosed left anterior descending segments but not in segments proximal or distal to the thrombus site, indicating that the vascular wall per se may be a source of increased production of TXA2.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427118 TI - Influence of thrombolytic therapy on the evolution of baroreflex sensitivity after myocardial infarction. AB - Depressed baroreceptor sensitivity (BRS) has been associated with an increased risk of ventricular arrhythmias and sudden cardiac death after myocardial infarction, but the influence of thrombolytic therapy on BRS has not been examined. To determine the effect of thrombolytic therapy on the evolution of BRS after myocardial infarction, BRS was assessed at 6 days, 6 weeks, and 3 months in 76 patients, 53 (70%) of whom had received thrombolytic therapy. The mean age (57 vs 57 years), sites of infarction, and the proportion of patients taking beta blockers (68% vs 52%) did not differ between patients who did and those who did not receive thrombolytic therapy. There was no difference in predischarge mean left ventricular ejection fractions (42% vs 46%) between the two groups of patients, but mean baseline BRS was 9.2 (0.8) msec/mm Hg in patients who were treated with thrombolysis and 5.9 (1.3) msec/mm Hg in those who were not (p = 0.03). At 6 weeks the corresponding values were 9.7 (1.1) and 11.1 (2.8) msec/mm Hg (p = 0.6) and at 3 months 9.1 (1.0) and 6.5 (1.1) msec/mm Hg (p = 0.07). At baseline 13% of patients who were treated with thrombolysis and 13% of those who were not had BRS < 3.0 msec/mm Hg, but at 3 months 9% of patients who were treated with thrombolytic agents compared with 17% of those who had BRS < 3.0 msec/mm Hg. In conclusion, early after myocardial infarction mean BRS was higher in patients treated with thrombolysis compared with nontreated patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427119 TI - Estimation of anterior infarct size with body surface QRST integral maps in the presence of abnormal ventricular activation sequence in dogs. AB - The possibility of estimating infarct size with body surface QRST integral (IQRST) maps was investigated in dogs. IQRST maps were constructed from 87-lead body surface ECGs, which were recorded 1 week after the production of anterior myocardial infarction during artificial pacing that simulated normal conduction, left bundle branch block, and Wolff-Parkinson-White syndrome in 11 dogs. Small differences were observed between the IQRST maps of the normal conduction and left bundle branch block models (r = 0.93, root mean square difference = 8.71 mVmsec) and between the normal conduction and Wolff-Parkinson-White models (r = 0.96, root mean square difference = 6.03 mVmsec). Summation of the QRST integral values over the body surface leads (QRST index) inversely correlated with infarct size in all three conductions models: r = 0.91 (p < 0.001) in the normal conduction model; r = -0.81 (p < 0.001) in the left bundle branch block model; and r = -0.86 (p < 0.001) in the Wolff-Parkinson-White model. These results show that IQRST maps permit noninvasive estimation of infarct size, even in the presence of abnormal activation sequences. PMID- 8427120 TI - Supported angioplasty with synchronized retroperfusion in high-risk patients with left main trunk or near left main trunk obstruction. AB - To test the feasibility of synchronized retroperfusion (SRP) as a support device of percutaneous transluminal coronary angioplasty (PTCA) for high-risk patients, 10 patients with left main trunk or near left main trunk obstruction underwent PTCA with SRP. An 8.5F retroperfusion catheter was inserted from the antecubital vein into the coronary sinus. Arterial blood was supplied through the catheter into the myocardium with a retroperfusion pump during the diastolic phase by means of ECG triggering. In all patients, the narrowings were successfully dilated and an improvement of more than 20% in the luminal diameter stenosis was achieved; however, narrowing of more than 50% (58%) remained in one patient. In all patients, systemic hemodynamics was maintained for more than 30 seconds during balloon inflation. In seven patients, a 60-second balloon inflation was possible without any collapse of systemic hemodynamics. To test the protective effect of SRP on myocardial ischemia and impairment of systemic hemodynamics, balloon inflation without SRP was performed in eight patients after successful dilatation. The duration for balloon inflation with SRP (71 +/- 30 seconds; n = 8) was significantly longer than that without SRP (56 +/- 30 seconds; n = 8). The decrease in systolic aortic pressure, the increase in pulmonary diastolic pressure, and ST-T segment elevation in the precordial lead of ECG during balloon inflation with SRP were less than those during balloon inflation without SRP. After PTCA, angina was not provoked by exercise stress testing in any of the 10 patients. We concluded that SRP is a beneficial support device of PTCA for high risk patients. PMID- 8427121 TI - Matching based on quantitative coronary angiography as a surrogate for randomized studies: comparison between stent implantation and balloon angioplasty of native coronary artery lesions. AB - Although intracoronary stenting has been advocated as an adjunct to balloon angioplasty to circumvent late restenosis, its effectiveness has not yet been verified. Therefore the aim of this study was to determine the differences in the immediate and long-term changes in stenosis geometry between Wallstent implantation and balloon angioplasty in native coronary artery lesions. To obtain two study populations with identical baseline stenosis characteristics, patients were matched for lesion site, vessel size, and minimal luminal diameter. Only patients undergoing elective and successful coronary intervention of a native coronary artery, in whom a control angiographic study had been performed, were included. A total of 186 patients (93 in each group) were selected. The coronary angiograms were analyzed with the computer-assisted cardiovascular angiographic analysis system. Matching was considered adequate, since there was an equal number of lesion sites in each study population and there were no differences in baseline reference diameter and minimal luminal diameter. Wallstent implantation resulted in a significantly greater increase in minimal luminal diameter (from 1.22 +/- 0.34 mm to 2.49 +/- 0.40 mm, p < 0.00001) compared with balloon angioplasty (from 1.21 +/- 0.29 mm to 1.92 +/- 0.35 mm, p < 0.00001). Despite a greater decrease in minimal luminal diameter after Wallstent implantation (0.48 +/- 0.74 mm) than after balloon angioplasty (0.20 +/- 0.46 mm), the minimal luminal diameter at follow-up was significantly greater after stent implantation (2.01 +/- 0.75 mm vs 1.72 +/- 0.54, p < 0.0001). It was concluded that Wallstent implantation results in a superior immediate and long-term increase in minimal luminal diameter compared with balloon angioplasty. The larger initial gain after stent implantation compensates for the late loss, and thus an improved initial result and not lessened neointimal hyperplasia is responsible for a reduced incidence of restenosis. Studies based on matching of angiographic variables are a surrogate for randomized studies, forecasting their results and offering insight into the effects of different interventional techniques. Moreover, these studies yield statistical information that may be helpful for the proper design of a randomized study (sample size, type II error). PMID- 8427122 TI - Low serum apolipoprotein A-I level in patients with vasospastic angina. AB - The serum lipid and lipoprotein levels in 15 patients with vasospastic angina were compared with those in 33 patients with no angiographic coronary stenosis and no vasospastic angina after intracoronary acetylcholine infusion. The serum level of apolipoprotein A-I in patients with vasospastic angina (112 +/- 6 mg/dl) was significantly lower (p < 0.05) than that in patients without vasospasm (128 +/- 4 mg/dl). However, there were no differences between the two groups in the serum levels of cholesterol, triglycerides, high-density lipoprotein cholesterol, apolipoprotein A-II, and apolipoprotein B. Thus apolipoprotein A-I may play some role in the prevention of vasospastic angina. PMID- 8427123 TI - Assessment of autonomic nervous activity by heart rate spectral analysis in patients with variant angina. AB - The purpose of this study was to assess the role of the autonomic nervous system in the pathogenesis of coronary artery spasm in patients with variant angina. We evaluated cardiac sympathetic and parasympathetic activity from the power (logarithmic scale) of the low-frequency (approximately 0.04 to 0.12 Hz) and the high-frequency (approximately 0.22 to 0.32 Hz) spectral components of heart rate variability with Holter monitoring in seven patients with nocturnal variant angina and in 11 healthy men who served as control subjects. None of the patients had organic coronary artery stenosis as determined by angiography. Low-frequency and high-frequency logarithmic values were calculated for each 5-minute period from 30 minutes before to immediately before each angina attack. The logarithmic low-frequency value during the 5-to-0-minute period was greater than the low frequency values during most of the other periods (p < 0.05 - p < 0.01). The logarithmic high-frequency values during the 10-to-5-minute and 5-to-0-minute periods were greater than those during the 30-to-25-minute period (p < 0.05 and p < 0.01, respectively). These data indicate that parasympathetic activity increased during the 10 minutes before attacks of nocturnal variant angina, whereas sympathetic activity with vagal modulation increased during the 5 minutes before such attacks. The same pattern of changes in heart rate variability was found in the absence of ST-segment elevation in patients and in control subjects. So this phenomenon was not just associated with coronary spasm and variant angina. It is suggested that circadian variation in disease activity is also associated with spontaneous attacks. PMID- 8427124 TI - Reinjection as an alternative to rest imaging for detection of exercise-induced ischemia with thallium-201 emission tomography. AB - Exercise thallium-201 single photon emission computed tomography images were compared prospectively with 4-hour redistribution images, with 4-hour reinjection images, and with images obtained at rest on a separate day in 37 patients with documented coronary artery disease. Exercise images were abnormal in 35 patients (95%). On the basis of an improvement in thallium-201 distribution between exercise and nonexercise images, overall sensitivity for the detection of coronary artery stenosis was significantly higher with reinjection at 4 hours (p < 0.05) or with a rest injection on a separate day (p < 0.05) than with redistribution imaging (84%, 83%, and 70%, respectively). Reinjection and rest injection were positive more frequently in patients with a wall-motion abnormality (76% and 80%, respectively, vs 64% at redistribution; p < 0.05 for both) or with > 90% stenosis (77% and 76%, respectively, vs 58% at redistribution; p < 0.05 for both). Among the 11 patients who had no evidence of redistribution at 4 hours, five (45%) demonstrated ischemia with reinjection and five demonstrated ischemia in the separate rest study; a total of seven patients showed improvement either at reinjection or rest. Among these 86% had a wall motion abnormality associated with stenosis of > 90%, whereas in the other 30 patients these two conditions were observed concomitantly in only 43%. This study demonstrates that the thallium-201 4-hour postexercise reinjection technique is as sensitive as the 2-day rest/exercise method for the detection of coronary artery stenosis and provides additional information when a severe stenosis is associated with a wall-motion abnormality. PMID- 8427125 TI - Variables associated with a poor prognosis in patients with an ischemic thallium 201 exercise test. AB - To determine the exercise workload, ECG, and thallium-201 image parameters that are most closely associated with a poor prognosis from ischemic heart disease, the test results of 268 patients were reviewed. Only patients with unequivocal thallium-201 redistribution were selected. A multivariate analysis was performed to find the variables that were most strongly associated with the outcomes of coronary revascularization, myocardial infarction, and cardiac death during a follow-up period of 25 +/- 19 months. Patients who underwent early elective revascularization had poorer exercise tolerance and more thallium image abnormalities than those with no events. In the remaining patients myocardial infarction was most closely related to the extent and severity of thallium ischemia (p = 0.0086), whereas cardiac death was associated with abnormal thallium lung uptake (p = 0.0082) and an inability to exercise to 9.6 MET (p = 0.0144). Thus unlike myocardial infarction, cardiac death is best predicted by variables that reflect poor left ventricular function rather than those that indicate ischemia. PMID- 8427126 TI - Effect of calcification and formalin fixation on in vitro distensibility of human femoral arteries. AB - To determine the effect of calcification on the distensibility of human arteries, we measured changes in lumen cross-sectional area as a function of pressure in excised segments of calcified and noncalcified human femoral arteries with the use of intravascular ultrasound. Cross-sectional area at zero pressure was greater in calcified artery segments (22.1 +/- 4.4 mm2) compared with noncalcified arteries (18.9 +/- 2.4 mm2). Calcified arteries were significantly less distensible than noncalcified arteries (2.5-fold). To assess the validity of mechanical tests of fixed arteries, pressure-area relations were also obtained after formalin fixation: mean unstressed cross-sectional area decreased significantly in noncalcified arteries (24%) but did not change significantly in calcified arteries. Formalin fixation decreased distensibility significantly (68%) in noncalcified arteries but not in calcified arteries. These results indicate that decreased lumen size and increased rigidity are induced by formalin fixation in noncalcified arteries but not in calcified arteries, and that this is attributable to the lower distensibility of calcified arteries before fixation. Because histologic examination is commonly used to evaluate severity of lumen narrowing in studies of atherosclerosis, this differential effect of formalin fixation may cause calcified arteries to appear less severely obstructed than noncalcified arteries. PMID- 8427127 TI - Effect of stenosis inlet geometry on shear rates of blood flow in the upstream region. AB - Abnormal shear rates of blood flow have been implicated in the processes of thrombosis, atherosclerosis, and restenosis after angioplasty. However, no study has quantitated the effect of stenosis inlet geometry on the shear rates in the region upstream to the stenosis. To quantitate this effect, we measured the velocity profiles of blood flow at Reynolds numbers 175 and 350 upstream to different axisymmetric model stenoses in an excised canine aorta. Two 40% and two 75% stenoses were tested, one each with a 45-degree inlet angulation and one each with a 90-degree angulation. For the velocity measurements we used a specially developed external single-channel Doppler ultrasound system capable of resolving blood flow velocity at 91 microns radial intervals across the aorta. We found that increasing the severity of stenosis narrowing and angulation resulted in a significant decrease in shear rate at the endothelial surface (40%/45-degree stenosis: 189 +/- 46 sec-1 vs 75%/90-degree stenosis: 49 +/- 12 sec-1 at Reynolds number 350; p < 0.002) and a significant increase in the maximum shear rate within the vessel lumen (189 +/- 46 sec-1 vs 295 +/- 8 sec-1, respectively; p < 0.05) in the region immediately upstream to the stenosis. These effects were less pronounced for Reynolds number 175. We conclude that stenosis inlet geometry has a significant impact on the flow conditions in the region immediately upstream to the stenosis, which is dependent on the Reynolds number. This may be an important determinant of thrombosis and atherogenesis in this particular region. PMID- 8427128 TI - Percutaneous transluminal venous angioplasty in occlusive iliac vein thrombosis resistant to thrombolysis. AB - Systemic thrombolysis is less than optimal in total occlusions of the iliac vein in which patency is 20% or less. We describe an interventional therapeutic procedure that may be effective in such cases. We selected 18 patients (average age, 29.5 years; range, 16 to 71 years) with complete iliac vein occlusion that persisted after 24 to 48 hours of systemic thrombolysis (streptokinase 100,000 U/hr). The ipsilateral femoral vein was punctured, and a guide wire was gently advanced through the thrombus into the inferior vena cava. Multiple inflations were performed with a balloon catheter that was advanced on the wire. A temporary vena cava filter was placed as a protection against possible embolic migration. Systemic thrombolysis was administered for 24 to 48 hours. Control venography and pulmonary angiography were performed. Venography showed good recanalization in seven cases, incomplete recanalization in five cases, and failure in six cases. Patency was maintained for a long time (15.6 months). In conclusion, (1) percutaneous transluminal venous angioplasty is a valuable adjunct to systemic thrombolysis when the latter alone fails; (2) segmental flow and mechanical obstruction were the critical factors, since the pharmaceutical factors were held constant, and (3) a more aggressive incremental interventional strategy warrants consideration. PMID- 8427129 TI - Coagulation abnormalities associated with the use of anabolic steroids. AB - According to a number of recent reports, persons using anabolic steroids may be subject to an increased risk of thromboembolism. We evaluated the effect of anabolic steroid use on the coagulation and fibrinolytic systems of 16 male bodybuilders to determine whether alterations occurred that would predispose them to a hypercoagulable state. No attempt was made to regulate or guide steroid use. Paired blood samples, both with and without steroid use, were obtained from six individuals, and the remaining subjects provided single samples obtained either during steroid use or nonuse. No differences were noted in most parameters, but we did find a significant increase in protein C antigen (p = 0.008) and free protein S antigen (p = 0.015), with a decreased euglobulin lysis time (p = 0.021) during steroid use. We also found a reduction in total cholesterol levels (p = 0.035) during steroid use. At least some of these findings suggest an activated fibrinolytic state, a known effect of anabolic steroids. The results do not support the presence of a hypercoagulable state. If anabolic steroids do produce a thrombotic tendency, they may do so through alterations in other hemostatic mechanisms or changes in lipid fractions, or more sensitive coagulation assays may be required for detection. PMID- 8427130 TI - Phase image analysis of anomalous ventricular activation in pediatric patients with preexcitation syndromes or ventricular tachycardia. AB - This prospective study evaluated the accuracy of phase analysis of scintigraphic imaging in defining the site of earliest ventricular activation in pediatric patients with electrophysiologic disorders. Twenty patients (10.8 +/- 5.5 years) with preexcitation (n = 16) or ventricular tachycardia (VT) (n = 4) were independently evaluated by phase image analysis and endocardial catheter mapping. The earliest phase angle (contraction), which was common to three scintigraphic imaging planes during preexcited sinus rhythm or VT, was compared with the earliest retrograde atrial activation during reciprocating tachycardia or the origin of VT, as defined by catheter mapping. Phase analysis of earliest contraction was concordant with catheter mapping of electrical activation in all 13 free-wall accessory connections and in three of four patients with VT. Inconclusive definition of activation occurred only in paraseptal accessory connections or VT. In conclusion, phase analysis accurately defines anomalous ventricular activation that is due to free-wall accessory connections or VT. In patients with complex anatomy or small size, phase analysis allows noninvasive localization of the anatomic substrates of tachycardia. PMID- 8427131 TI - Recurrent conduction in accessory pathway and possible new arrhythmias after radiofrequency catheter ablation. AB - Radiofrequency catheter ablation was performed in 142 patients with 166 accessory pathways. One hundred thirty-six patients with 160 accessory pathways underwent successful ablation in the first ablation session. Serial follow-up electrophysiologic studies were performed immediately (30 minutes), early (5 to 7 days), and late (3 to 6 months) after successful ablation to determine the recurrent accessory pathway conduction and possible new arrhythmias. After a minimum follow-up period of 6 months (mean, 14 +/- 3 months), accessory pathway conduction recurred in 13 patients (9.6%), with recurrent tachycardia in three patients (2.2%). Five of the recurrent accessory pathways had decremental conduction properties. Incidence of recurrent accessory pathway conduction was similar in different accessory pathway locations (6.4% to 9.0%). Patients with concealed accessory pathways (12.2 vs 2.9%; p < 0.05), and patients without accessory pathway potentials in the ablation site (15.5% vs 2.2%; p < 0.05) had a higher recurrence rate. Patients without tachycardia in the late electrophysiologic study did not have recurrent tachycardia during follow-up. New arrhythmias, including atrial and ventricular arrhythmias, which were detected by 24-hour Holter monitoring, were apparent only on the first day after ablation. The findings indicate that the overall incidence of recurrent accessory pathway conduction was low and that possible new arrhythmias were rare in the late follow up period. PMID- 8427132 TI - Arrhythmogenicity of catheter ablation in supraventricular tachycardia. AB - To evaluate arrhythmogenicity in patients who receive a modified direct-current (DC) shock ablation (distal pair of electrodes connected in common as the cathode) or radiofrequency (RF) ablation of supraventricular tachycardia, a prospective study was performed with signal-averaged ECG, 24-hour Holter monitoring, electrophysiologic study (EPS) for ventricular tachycardia (VT), and treadmill exercise test. Sixty-nine consecutive patients with documented paroxysmal supraventricular tachycardia were included. Twenty-eight patients proved to have atrioventricular nodal reentrant tachycardia, and 41 patients had atrioventricular reciprocating tachycardia that involved accessory atrioventricular pathways. The first 34 patients received DC shock ablation and the other 35 patients received RF ablation. Signal-averaged ECG, Holter monitoring, and EPS for VT were performed before ablation, immediately after ablation, then 1 week, 2 weeks (Holter monitoring), 1 month (except EPS), and 3 months after ablation. Treadmill exercise testing was performed before ablation, and at 1 week and 3 months after ablation. The root mean square, low-amplitude signal and QRS duration of signal-averaged ECG disclosed no significant change after either DC or RF ablation up to 3 months. Late potential developed in only one patient in the DC shock group and it was considered to be innocuous because neither VT nor ventricular fibrillation was noted or induced. Increases in the number of ventricular premature contractions and in short-run VT were detected by Holter monitoring in the first week after either mode of ablation (p < 0.001 for the DC shock group; p < 0.05 for the RF group), which were greater (p < 0.05) and lasted longer in the DC shock group than in the RF group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427133 TI - Transesophageal cardioversion. AB - With the use of a novel quadripolar esophageal electrode system, we have attempted 131 transesophageal cardioversions in 105 patients: 109 episodes were atrial fibrillation, 16 episodes were atrial flutter, 2 episodes were supraventricular tachycardia, and 4 episodes were ventricular tachycardia. The mean predicted transesophageal impedance (+/- SEM) of 52.6 +/- 1.1 omega was significantly lower than the mean predicted transthoracic impedance (+/- SEM) of 63.1 +/- 1.6 omega (n = 104; p < 0.01). Of the 88 patients who presented with atrial fibrillation as the initial arrhythmia, successful transesophageal cardioversion (maximal delivered transesophageal energy of 100 J in 84 patients and 200 J in 4 patients) was recorded in 70 (79.5%); transesophageal cardioversion required a mean delivered energy of 63.1 +/- 4.2 J and a mean peak current of 20.3 +/- 0.6 A. Transthoracic countershock (maximal delivered energy of 360 J) was attempted in 17 of 18 patients when the transesophageal approach had been unsuccessful; countershock was successful in 10 patients, which yielded an overall success rate of 92.0% (mean successful delivered energy [transesophageal and transthoracic] of 85.3 +/- 7.8 J). All episodes of atrial flutter, supraventricular tachycardia, and ventricular tachycardia were successfully terminated with the use of the esophagus. This esophageal electrode system permits low-energy countershock of atrial and ventricular tachyarrhythmias. PMID- 8427134 TI - Biphasic versus sequential pulse defibrillation: a direct comparison in humans. AB - It has been demonstrated recently that both biphasic and sequential pulse defibrillation shocks are superior to monophasic defibrillation shocks in animals and humans. There is little information directly comparing these two waveforms when pulse characteristics, subjects, and total electrode surface areas are kept constant. We determined the defibrillation threshold intraoperatively in 12 patients undergoing arrhythmia surgery, with the use of two or three patch electrodes (Medtronic 6891 and 6892), while keeping the electrode surface area constant. Patients were randomized in a crossover design for determinations of defibrillation threshold by means of biphasic and sequential pulse shocks. Leading-edge delivered current and delivered energy were significantly lower with sequential pulse shocks than with biphasic shocks (delivered energy means +/- SEM 3.6 +/- 0.7 joules vs 5.5 +/- 0.9 joules, respectively). We conclude that sequential pulse defibrillation with three defibrillating electrodes provides an important current delivery system not matched by biphasic shocks with two electrodes when subjects, waveform characteristics, and total electrode surface areas are kept constant. PMID- 8427135 TI - The spectrum of left ventricular size in dilated cardiomyopathy: clinical correlates and prognostic implications. SPIC (Italian Multicenter Cardiomyopathy Study) Group. AB - To address the issues of variability and prognostic role of left ventricular dimensions in dilated cardiomyopathy (DCM), 144 patients with DCM were studied. They were arbitrarily assigned to two groups according to an echocardiographic left ventricular end-diastolic diameter index < or = 15% (45 patients with mildly dilated cardiomyopathy) and above 15% (99 patients with typically dilated cardiomyopathy) of the upper normality range. Among the patients with mildly dilated cardiomyopathy, there were more men (89% vs 66%; p < 0.01). This group of patients also had a greater prevalence of atrial fibrillation (22% vs 3%; p < 0.001) higher left ventricular fractional shortening (15 +/- 6% vs 13 +/- 5%; p < 0.05), higher ejection fraction (28 +/- 8% vs 24 +/- 8%; p < 0.01), and a lower exercise tolerance (5 +/- 2 MET vs 6 +/- 2 MET; p < 0.05). At the time of follow up examination (30 +/- 15 months), event-free survival was not significantly different between patients with mildly dilated cardiomyopathy and those with typically dilated cardiomyopathy. Pulmonary capillary wedge pressure (p < 0.001) and left atrial dimension index (p < 0.01) were significant predictors of prognosis as determined by Cox multivariate analysis. Minimal or mild ventricular dilatation is not uncommon in DCM, and it identifies a heterogenous group of patients--some who are in the early stages of disease and others with severe pump dysfunction and persistently small hearts. Ventricular dilatation is not an independent predictor of prognosis. PMID- 8427136 TI - Long-term outcome of enoximone therapy in patients with refractory heart failure. AB - Few options are available for patients with severe heart failure that is unresponsive to therapy with digoxin, diuretics, and vasodilators. The clinical responses and predictors of survival were studied in 41 consecutive patients with New York Heart Association (NYHA) class IV heart failure during long-term oral enoximone therapy (mean dose 232 +/- 15 mg/day). The mean age was 60 +/- 1 years, and the initial left ventricular ejection fraction was 0.19 +/- 0.01. The cause of heart failure was either coronary artery disease (n = 23) or dilated cardiomyopathy (n = 18). Symptomatic improvement occurred in the majority (83%) of patients; 24% improved two or more NYHA classes. Although the 12-month mortality rate for the entire group was high (54 +/- 8%), a subgroup of patients with dilated cardiomyopathy achieved a sustained benefit with a decrease in symptoms > 1 NYHA class, fewer hospitalizations, and a survival rate at 24 months of 60%. Multivariate analysis identified the cause of heart failure, left ventricular ejection fraction, and clinical improvement within 60 days of enoximone therapy as predictors of a favorable long-term outcome. The presence of coronary artery disease was most predictive of early mortality (p < 0.0002), with only 5% of patients surviving > 18 months compared to 66% of those with dilated cardiomyopathy. Median survival rates were 132 +/- 31 and 921 +/- 214 days (p < 0.001) for the coronary artery disease and dilated cardiomyopathy populations, respectively. Oral enoximone can provide symptomatic improvement and a palliative option for the majority of patients with refractory heart failure resulting from cardiomyopathy. PMID- 8427137 TI - Hemodynamic and hemorheologic determinants of left atrial spontaneous echo contrast and thrombus formation in patients with idiopathic dilated cardiomyopathy. AB - The purpose of the present study was to evaluate the specific role of hemorheologic and hemodynamic parameters for spontaneous echo contrast and thrombus formation in vivo. We therefore investigated the association between the presence of left atrial spontaneous echo contrast and thrombus formation by transesophageal echocardiography and multiple clinical, hemodynamic, and hemorheologic parameters in 70 patients with idiopathic dilated cardiomyopathy. Transesophageal echocardiography showed left atrial spontaneous echo contrast and left atrial thrombi in 33% and 19% of patients, respectively. Patients with left atrial spontaneous echo contrast had a lower cardiac index (2.1 +/- 0.9 versus 2.6 +/- 0.9 L/min/m2; p < 0.02), a lower left atrial (21 +/- 8 versus 38 +/- 10 cm/sec; p < 0.001) and left atrial appendage flow velocity (17 +/- 14 versus 39 +/- 13 cm/sec; p < 0.001), a larger left atrial diameter (53 +/- 6 versus 46 +/- 10 mm; p < 0.002), and more often presented with atrial fibrillation (62% versus 32%; p < 0.02). Plasma fibrinogen concentration (4.0 +/- 1.1 versus 3.5 +/- 0.7 gm/L; p < 0.02) and plasma viscosity (1.83 +/- 0.10 versus 1.76 +/- 0.15 mPa.sec; p < 0.05) were higher in patients with spontaneous echo contrast. Multivariate analysis revealed an association between the presence of spontaneous echo contrast and left atrial flow velocity p < 0.0001) and plasma viscosity (p < 0.01). In patients with left atrial (appendage) thrombus or a history of embolism, left atrial appendage flow velocity was lower (15.0 +/- 8.2 versus 29.6 +/- 14.5 cm/sec; p < 0.005) and spontaneous echo contrast was more frequently observed (52% versus 23%; p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427138 TI - Diastolic dysfunction in cardiac transplant recipients: an important role in the response to increased afterload. AB - We evaluated the hemodynamic and functional response to acute elevations in left ventricular (LV) afterload in 22 recent recipients of cardiac transplants to determine whether abnormalities in LV diastolic function influence the response to this intervention. In seven patients (group 1) LV ejection fraction decreased significantly from baseline values (> or = 5%) during methoxamine infusion, whereas in 15 patients (group 2) LV ejection fraction was maintained. Peak filling rate was lower in group 1 versus group 2 (3.36 +/- 0.46 vs 4.23 +/- 0.68 end-diastolic volumes/sec, p < 0.01). In addition, patients in group 1 did not have LV dilatation during methoxamine (percentage change in end-diastolic counts, -3.4 +/- 6.9%) and had a large increase in pulmonary artery wedge pressure. In contrast, patients in group 2 had LV dilatation (percentage change in end diastolic counts, +10.7 +/- 14.7%) and a smaller increase in pulmonary artery wedge pressure. There was a relationship between the baseline peak filling rate and the change in LV ejection fraction during methoxamine (r = 0.65, p = 0.001). Therefore in a subset of cardiac transplant patients, abnormalities in LV filling can have an impact on the response to increased afterload. PMID- 8427139 TI - The angle of incidence of the ultrasonic beam: a critical factor for the image quality in intravascular ultrasonography. AB - The effects of the angle of incidence of the ultrasound beam on the image quality were studied in 21 pressurized arterial specimens examined with a 30 MHz intravascular ultrasonographic catheter. When the ultrasonographic catheter was in an eccentric position in the vessel lumen, the videodensity of the segments of the vessel wall with the least favorable angle of interrogation (a shift of 49 +/ 6 degrees from the tangent to the tissue surface) was 27% +/- 19% lower than the videodensity measured with the catheter in the center of the lumen. When the catheter was placed in a position that was not parallel to the long axis of the vessel, a further decrease was observed, especially in the vessel wall opposite the position of the catheter. An artificial dissection was induced in eight specimens. Dropouts that involved the dissection plane and the underlying structures were produced with positions of the echographic catheter inducing a narrow angle between ultrasound beam and dissection plane. These experimentally induced artifacts were compared with similar findings from the in vivo evaluation of peripheral and coronary arteries. The angle of incidence of the ultrasound beam is a major determinant of the image quality in intravascular ultrasonography. Angle-dependent artifacts occur with eccentric and noncoaxial positions of the ultrasonographic catheter and, in particular, with imaging of large intraluminal dissections. Awareness of this problem may prevent image misinterpretation and has relevance for future improvement of catheter technology and design. PMID- 8427140 TI - Accuracy of flow convergence estimates of mitral regurgitant flow rates obtained by use of multiple color flow Doppler M-mode aliasing boundaries: an experimental animal study. AB - The proximal flow convergence method of multiplying color Doppler aliasing velocity by flow convergence surface area has yielded a new means of quantifying flow rate by noninvasively derived measurements. Unlike previous methods of visualizing the turbulent jet of mitral regurgitation on color flow Doppler mapping, flow convergence methods are less influenced by machine factors because of the systematic structure of the laminar flow convergence region. However, recent studies have demonstrated that the flow rate calculated from the first aliasing boundary of color flow Doppler imaging is dependent on orifice size, flow rate, aliasing velocity and therefore on the distance from the orifice chosen for measurement. In this study we calculated the regurgitant flow rates acquired by use of multiple proximal aliasing boundaries on color Doppler M-mode traces and assessed the effect of distances of measurement and aliasing velocities on the calculated regurgitant flow rate. Six sheep with surgically induced mitral regurgitation were studied. The distances from the mitral valve leaflet M-mode line to the first, second, and third sequential aliasing boundaries on color Doppler M-mode traces were measured and converted to the regurgitant flow rates calculated by applying the hemispheric flow equation and averaging instantaneous flow rates throughout systole. The flow rates that were calculated from the first, second, and third aliasing boundaries correlated well with the actual regurgitant flow rates (r = 0.91 to 0.96). The mean percentage error from the actual flow rates were 151% for the first aliasing boundary, 7% for the second aliasing boundary, and -43% for the third aliasing boundary; and the association between aliasing velocities and calculated flow rates indicates an inverse relationship, which suggests that in this model, there were limited velocity-distance combinations that fit with a hemispheric assumption for flow convergence geometry. The second aliasing boundary with an aliasing velocity, of 102 cm/sec, (which was achieved by use of a 4 kHz pulse repetition frequency, a 3.75 MHz transducer, and no color baseline shift), provided the closest fit to the actual regurgitant flow rates (r = 0.99; y = 0.95x + 0.07). The averaged calculated flow rates from all aliasing velocities also resulted in excellent correlation (r = 0.97; y = 0.99x + 0.5). A hemispheric flow convergence method that is based on color Doppler M-mode echocardiography is a feasible and automatable method for quantifying mitral regurgitant rate.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8427141 TI - Two-dimensional echocardiographic anatomy of atrioventricular alignment discordance with situs concordance. AB - Relationships between the atria and the ventricles are commonly described in terms of either atrioventricular (AV) alignments (or connections) or AV situs (i.e., the type of atrial situs and the type of ventricular situs or ventricular loop). With either method of analysis, only one type of AV relationship (situs or alignment) is diagnosed specifically and is considered to be predictive of the other type of AV relationship. The two-dimensional echocardiographic characteristics of two patients with incongruent AV situs and alignments are described for the first time. Patient 1 had situs solitus of the viscera and atria (S), ventricular D-loop (D), and solitus normally related great arteries (S), or (S,D,S) segmental combination with concordant AV situs. However, the right-sided right atrium drained into the inferior and right-sided left ventricle, and the left-sided left atrium drained into the superior and left sided right ventricle. Hence AV alignment discordance was present. Patient 2 had visceroatrial situs solitus (S), ventricular D-loop (D), and double-outlet right ventricle with a rightward aortic valve (D), or (S,D,D) segmental set (AV situs concordance). Similar to patient 1, AV alignment discordance was demonstrated. In both patients the diagnosis was established by two-dimensional echocardiography and subsequently confirmed by cardiac catheterization. The key to accurate echocardiographic diagnosis of this congenital heart disease was independent analysis of the AV alignments and the segmental situs of the three main cardiac segments. This diagnosis was determined by scanning from the parasternal, subxiphoid, and apical windows.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427142 TI - Repair of coarctation of the aorta in infancy: comparison of surgical and balloon angioplasty. AB - Surgical repair of coarctation of the aorta in infancy has recently been challenged by some investigators who suggest that balloon angioplasty results in a lower mortality rate and similar risk of restenosis. Over a 44-month period, 37 consecutive infants with a mean age of 33 days (median, 15 days; range, 1 to 200 days) and mean and median weight of 3.7 kg (range, 2.4 to 5.4 kg) underwent surgical repair of coarctation of the aorta with either an end-to-end anastomosis (24 patients) or subclavian flap angioplasty (13 patients). There were no operative deaths (95% confidence interval, 0% to 10%). Four patients died late (> 30 days) after surgery (11%). Four patients (11%) (95% confidence interval, 3% to 25%) had residual gradients greater than 20 mm Hg. A review of the recent literature on treatment of native coarctation in infants with surgical repair (18 reports, 1189 patients) and balloon angioplasty (8 reports, 57 patients) reveals a similar early mortality rate but a much higher rate of recoarctation in infants who were treated with balloon dilation (57%) as compared with those who underwent surgical repair (14%). Because of the incidence of restenosis, balloon dilation as compared with surgical repair does not yet offer an improved outcome for native coarctation of the aorta in infancy. PMID- 8427143 TI - Management of cocaine-induced cardiovascular complications. PMID- 8427144 TI - The head-up tilt table test and cardiovascular neurogenic syncope. PMID- 8427145 TI - Diet, menopause, and serum cholesterol levels in women: the Framingham Study. AB - Cross-sectional relationships between diet and total serum cholesterol levels were studied in a sample of 428 women from the Framingham Heart Study Cohort, aged 37 to 70 years, from 1957 to 1960. Multiple linear regression was used to control for total calorie intake, systolic blood pressure, physical activity, Metropolitan relative weight, glucose intolerance, and cigarette smoking. There was little evidence for a relationship between total serum cholesterol and dietary fat intake; whereas a marginally significant direct association was found with total fat in postmenopausal women, total and plant fat and cholesterol were inversely associated, and only cholesterol was significant in premenopausal women. A consistent inverse association was observed between total serum cholesterol levels and intake of protein, particularly from plant sources, and a weak inverse association was found with complex carbohydrate intake. Serum cholesterol in women may be influenced by a number of dietary factors and appears to differ according to menopausal status. PMID- 8427146 TI - Preliminary observations on the use of the Palmaz stent in the distal portion of the abdominal aorta. AB - The opportunities to avoid surgical treatment of distal abdominal aortic occlusive disease are expanding because of the proliferation in catheter-based techniques. Since January 1990, 24 symptomatic patients with distal abdominal disorders have been treated percutaneously with balloon angioplasty and intraluminal stenting. A total of 38 Palmaz stents were deployed at distal abdominal aortic sites; 21 additional iliac stents were implanted. After the procedure all patients improved clinically, and 83% (21/24) improved by objective measurement (average ABI 0.93 +/- 0.21). Three access-related complications occurred (two hematomas, one thrombus), but no complications were related to the stents. At up to 29 months of follow-up (average 10.3 +/- 6.7 months), clinical improvement persists in all patients (average ABI 0.93 +/- 0.22). In 11 patients eligible for follow-up aortography, all aortic stents are patent without evidence of restenosis. With the low likelihood for restenosis in the abdominal aorta, the long-term outcome of aortic stenting looks promising and may equal or surpass that found in the iliac region. PMID- 8427147 TI - A new expandable intracoronary tantalum (Strecker) stent: early experimental results and follow-up to twelve months. AB - A new radiopaque balloon expandable tantalum stent was tested in the coronary arteries of sheep. A total of 28 stents with a diameter of 2.0 to 3.8 mm were successfully deployed. The stent to coronary artery diameter ratio was 1.1-1.2:1. The animals were heparinized with 100 U/kg of heparin but did not receive antiplatelet drugs. Coronary angiography that was performed 10 minutes after stent implantation showed 100% patency with no side-branch occlusion. Four sheep died within 2 hours of stent implantation, and pathologic studies showed thrombosis of the smaller sized stents: 2 mm (n = 2), 2.8 mm (n = 1), and 3.2 mm (n = 1); three fourths of the sheep had two stents implanted. An oversized stent caused coronary rupture and cardiac tamponade in one other sheep. Follow-up study protocol included coronary angiography before animal sacrifice and pathologic studies within 48 hours (n = 11), 2 weeks (n = 1), 3 months (n = 2), 5 months (n = 1), 10 months (n = 7), and 12 months (n = 1). At 3 to 12 months of follow-up the coronary stent was completely covered with a layer of neointima, and there was no angiographic evidence of coronary stenosis and patent side branches. According to histologic examination, the neointima had nonuniform thickness (20 to 330 microns) and consisted of smooth muscle cells and some collagen. At 7 months of follow-up one of seven stents had angiographic (20%) and pathologic evidence of stenosis, which was secondary to thrombus. The tantalum device is easily deployed technically and notable for its excellent fluoroscopic visibility and flexibility. Thrombosis with a tantalum stent remains a problem, and therefore the larger diameter stents > or = 3.2 mm, antiplatelet drugs, and anticoagulation therapy are indicated for human studies. Implantation of multiple coronary stents increases the risk of stent thrombosis. The long-term, mild neointimal thickness and the patency of the stent and side branches are encouraging. PMID- 8427148 TI - Excimer and holmium yttrium aluminum garnet laser coronary angioplasty. AB - Recently access to the coronary arteries became available to laser angioplasty because of a new technique utilizes a pulsed laser source and multifiber, "over the-wire" guided catheters. The aim of this study was to evaluate the early and long-term results and the side effects of coronary angioplasty with an excimer or a Ho-YAG laser. Forty consecutive patients were treated with the Ho-YAG laser (group I) and 46 consecutive patients were treated with the excimer laser (group II). The primary laser angioplasty success rate was 55% and 72% (NS) for groups I and II, respectively. This success rate was highest in saphenous vein grafts. It was similar in calcified and noncalcified lesions and in total occlusions and stenoses. It tended to be lower in long lesions than in short ones (40% vs 60%; p < 0.05 and 44% vs 78%; NS for groups I and II, respectively). Laser stand-alone therapy was performed in 5% of patients in group I compared with 22% in group II (p < 0.05). Failures were due to the inability of the laser catheter tip to reach the lesion, to cross the obstruction, or to obtain a significant reduction in stenosis. They were more frequent in patients in group I than in those in group II (45% vs 28%). There were no deaths, no myocardial infarctions, and no need for emergency coronary artery bypass grafting because most patients had total occlusions or a well-protected coronary artery. Complications included acute closure in 8% of patients in group I and in 17% of patients in group II and spasm in 10% and 13% of patients in groups I and II, respectively. Dissection occurred more frequently in patients in group II than in those in group I (28% vs 7%; p < 0.04). The angiographic patency rate at 6-month follow-up was 33% and 29% for patients groups I and II, respectively. Multifiber, wire-guided catheters provide easy access to the coronary arteries. Excimer laser angioplasty is effective but induces a high rate of dissections. Technical improvements are required to ablate more tissue to possibly reduce the restenosis rate. Further studies are needed to elucidate the mechanism of side effects and to reduce the restenosis rate. PMID- 8427149 TI - Unusual variant of single left coronary artery. PMID- 8427150 TI - A rare variant of single left coronary artery. PMID- 8427151 TI - Doppler-tipped guide wire assessment of retrograde coronary artery flow distal to a total occlusion and its reversal after laser recanalization. PMID- 8427152 TI - Use of intraaortic balloon counterpulsation for treatment of recurrent acute closure after coronary angioplasty. PMID- 8427153 TI - Spontaneous rupture of papillary muscle due to remote myocardial infarction. PMID- 8427154 TI - Transesophageal echocardiography in the detection of left ventricular pseudoaneurysm. PMID- 8427155 TI - Myocardial perfusion imaging during stress-induced sustained ventricular tachyarrhythmia. PMID- 8427156 TI - Asymptomatic live systemic embolization observed on transesophageal echocardiography. PMID- 8427157 TI - Migration of a Kimray-Greenfield filter into the pulmonary artery: localization by transesophageal echocardiography. PMID- 8427158 TI - Acute valvular obstruction from streptococcal endocarditis. PMID- 8427159 TI - A new resting site for paradoxical thromboemboli: the mitral valve apparatus. PMID- 8427160 TI - The atrial septum after balloon mitral valvotomy: observations during surgery. PMID- 8427161 TI - Deletion of mitochondrial DNA in a patient with conduction block. PMID- 8427162 TI - Transesophageal echocardiographic imaging in the right lateral decubitus position is comparable to the conventional left lateral decubitus position. PMID- 8427163 TI - Angioscopic and intravascular ultrasound imagings before and after percutaneous holmium-YAG laser coronary angioplasty. PMID- 8427164 TI - Reducing lengths of stay for patients hospitalized with chest pain using medical practice guidelines and opinion leaders. AB - There are few available data on the effectiveness and safety of medical practice guidelines when used for patient care in the coronary and intermediate care units. The aim of this study was to examine the effect of educating physicians about practice guidelines to promote shorter lengths of stay for "low-risk" patients hospitalized with chest pain. Such guidelines were disseminated to physicians working in a health maintenance organization (HMO) by educational conferences, written memoranda, endorsement by opinion leaders, concurrent written feedback, and nursing-to-physician cues. A total of 208 patients were enrolled in the study. Following implementation of the practice guidelines, hospital lengths of stay were reduced from 2.51 +/- 2.1 to 1.96 +/- 1.3 days (22% reduction, p = 0.03) and intermediate care unit lengths of stay from 33.9 +/- 19 to 28.2 +/- 14 hours (17% reduction, p = 0.02) for patients with low-risk chest pain. The reduction in length of stay for patients with low-risk chest pain exceeded reductions in stay for patients hospitalized with cardiac conditions for which no guidelines were introduced. None of the patients treated according to guideline recommendations had unexpected "life-threatening" adverse events in the 2-week period after hospital discharge (95%, confidence interval 0%, 3%). This study supports the effectiveness and possible safety of practice guidelines to reduce lengths of stay for patients with low-risk chest pain. PMID- 8427166 TI - A community-wide perspective of gender differences and temporal trends in the use of diagnostic and revascularization procedures for acute myocardial infarction. AB - This study compares the overall use, as well as temporal trends, of various diagnostic and revascularization procedures for acute myocardial infarction (AMI) in men and women. The study sample comprised a total of 2,924 men and 1,838 women with validated AMI admitted to any of the 16 teaching and community hospitals in the Worcester, Massachusetts, metropolitan area during 1975, 1978, 1981, 1984, 1986 and 1988. During the period under study there was a significant increase in use of each of the examined procedures during hospitalization for AMI in both men and women. Increasing use of multiple procedures was also seen for each of the sexes. After controlling for a variety of demographic and clinical factors that might affect utilization rates, men were marginally more likely to undergo radionuclide ventriculography, and significantly more likely to undergo Holter monitoring, exercise treadmill testing, cardiac catheterization, and percutaneous transluminal coronary angioplasty than women. However, there were no gender differences in the use of coronary artery bypass grafting. On the other hand, men were significantly less likely to undergo echocardiography. The results of this multihospital, population-based study suggest sex differences in the use of several diagnostic and revascularization procedures during hospitalization for AMI. These differences may be attributed to physicians' practice patterns, although gender bias in the delivery of medical care cannot be excluded. Temporal trends in increased overall use of these procedures raise questions about cost effectiveness that need to be further addressed. PMID- 8427165 TI - Psychosocial predictors of mortality in the Cardiac Arrhythmia Suppression Trial 1 (CAST-1). AB - Psychosocial variables predict the recurrence of clinical events in symptomatic patients, controlling for measures of disease severity. The Cardiac Arrhythmia Suppression Trial-1, a pharmacologic test of the arrhythmia suppression and mortality hypothesis among postmyocardial infarction patients, allowed a prospective test of the relationship of distress, perceived support, social interaction, life stress, and other variables, to mortality, adjusting statistically for ejection fraction, arrhythmia rates, and other known risk factors for coronary heart disease. Results indicated that the treatment medications, encainide and flecainide, were powerful predictors of mortality. Although the psychosocial variables were significant as univariate predictors, these variables were not significant as predictors in a multivariate model that included drug treatment. When the data analysis was restricted to patients randomized to placebo, thereby eliminating the antiarrhythmic drug effect, the level of perceived social support was a significant multivariate predictor of mortality, adjusting for measures of disease severity. The adjusted hazards ratio for a 1-point decrease in the perceived support score is equal to 1.46, based on the multivariate model. PMID- 8427167 TI - Comparison of results of intravenous infusion of anistreplase versus streptokinase in acute myocardial infarction. AB - This randomized study compares the coronary perfusion rate in patients with acute myocardial infarction (AMI) treated with 2 different intravenous thrombolytic agents: streptokinase 1.5 million U given over 60 minutes and anisoylated human plasminogen streptokinase activator complex (anistreplase) administrated as a bolus of 30 U over 5 minutes. One hundred seventy-five patients (149 men and 26 women, mean age 54 years) have been included in this study. Eighty-nine patients were treated with anistreplase and 86 patients with streptokinase. AMI was inferior in 54 patients (61%) in the anistreplase group and in 54 patients (63%) in the streptokinase group. It was anterior in 35 (40%) and 32 (37%) patients, respectively. Coronary angiography and ventriculography were performed at a mean time (+/- SEM) of 207 +/- 11 minutes after the beginning of thrombolysis in 170 patients. A perfusion score grade of 2 or 3 according to the Thrombolysis in Myocardial Infarction trial was found in 63 patients (72%) in the anistreplase group and in 56 patients (68%) in the streptokinase group (p = NS). Severe bleeding occurred in 7 patients (8%) after anistreplase and in 6 patients (7%) after streptokinase. No cerebral hemorrhage occurred. Nine patients (5%) died during their hospital stay: 6 after anistreplase and 3 after streptokinase. It is concluded that intravenous administration of anistreplase or streptokinase is efficient and safe. Coronary patency 207 minutes after fibrinolysis, incidence of adverse events and mortality are similar in both groups. PMID- 8427168 TI - Effectiveness of captopril in reversing renal vasoconstriction after Q-wave acute myocardial infarction. AB - The purpose of this investigation was to study whether favorable renal effects might contribute to the influence of captopril in offsetting ventricular dilatation after infarction. Effective renal plasma flow and glomerular filtration rate were estimated by isotope injection methods in 20 patients on days 2, 7, 8, 42 and 180 after a first transmural anterior myocardial infarction. After measurements on day 7, patients were randomized to receive either captopril 25 mg 3 times daily (n = 10) or placebo (n = 10) for the remainder of the study. At baseline (day 7) there were no differences between the 2 treatment groups in radionuclide left ventricular ejection fraction, effective renal plasma flow, glomerular filtration rate or neurohormones. Left ventricular ejection fractions (40 +/- 4% [mean +/- 2 SD] at baseline) were higher in the captopril- than the placebo-treated patients on days 42 (p < 0.05) and 180 (p < 0.01) after infarction. Effective renal plasma flow became significantly higher at all time points after randomization in the captopril-treated group than in the placebo group (p < 0.001). A similar but lesser trend was observed for glomerular filtration rate. Plasma atrial natriuretic factor and aldosterone were significantly higher in the placebo group (p < 0.05). Renal hemodynamic indexes were directly correlated with and neurohumoral indexes inversely correlated with ejection fractions. In a second group of 12 patients with higher baseline ejection fractions (48 +/- 4%) after an inferior infarction, none of these beneficial effects of captopril were demonstrable. It is proposed that in the setting of left ventricular dysfunction after infarction, a prompt and sustained improvement in renal hemodynamics, by reducing inappropriate fluid retention and thus ventricular preload, may be one contributory mechanism by which captopril prevents progression of left ventricular dilatation. PMID- 8427169 TI - Physiologic responses to weight lifting in coronary artery disease. AB - This study assesses the safety of and physiologic responses to maximal repetition, dynamic, resistive weight lifting at 40, 60, 80 and 100% of maximal voluntary contraction compared with aerobic exercise using a maximal treadmill exercise test. Twelve men with coronary artery disease exercised to fatigue at 4 stations (overhead press, biceps curl, quadriceps extension and supine press). The electrocardiogram was monitored continuously. Heart rate and systolic and diastolic blood pressures (by sphygmomanometer) were measured at rest and during peak exercise. No symptoms or electrocardiographic evidence of ischemia occurred with weight lifting, whereas 5 of 12 patients had ischemic ST-segment depression (> or = 1 mm) with the treadmill. No significant ectopy occurred with either activity. Mean peak heart rates with all lifts were less (range 74 to 92 beats . min-1; p < or = 0.05) than with the treadmill (157 beats . min-1). Peak systolic blood pressures were similar, whereas peak diastolic blood pressures were greater with all lifts (range 93 to 117 mm Hg; p < or = 0.05), except 100% maximal contraction biceps curl and quadriceps extension, than with the treadmill (79 mm Hg). Peak rate pressure product was greater with the treadmill than with all lifts (p < or = 0.05). Diastolic time interval from the electrocardiograph was shorter with the treadmill (0.154 second) than with all lifts (range 0.323 to 0.448 second; p < or = 0.05). Diastolic pressure-time index was greater with all lifts than with the treadmill (p < or = 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427170 TI - Usefulness of supine bicycle stress echocardiography for detection of restenosis after percutaneous transluminal coronary angioplasty. AB - The role of supine bicycle stress echocardiography (SBSE) for detecting restenosis after percutaneous transluminal coronary angioplasty (PTCA) was evaluated in 80 patients: 41 (51%) with single and 39 (49%) with multivessel PTCA (total 129 dilated vessels). Total revascularization was performed in 54 (68%) and partial revascularization in 26 (32%) patients. Restenosis was angiographically demonstrated in 60 patients (75%) and in 72 vessels (56%) 6.1 +/ 2.9 months after PTCA. The results for detecting restenosis were: (1) SBSE versus exercise electrocardiographic sensitivity, 87 versus 55% (p < 0.001); (2) specificity, 95 versus 79%; and (3) accuracy, 89 versus 61% (p < 0.001). SBSE was 83% sensitive, 95% specific and 88% accurate for restenosis detection in specific vessels with comparable results for single versus multivessel PTCA and total versus partial revascularization. Sensitivity, specificity and accuracy were: 91, 93 and 91% for the left anterior descending coronary artery; 77, 94 and 85% for the right coronary artery; and 76, 96 and 88% for the left circumflex coronary artery. Ninety-four percent of the nondilated diseased vessels were correctly identified. It is concluded that SBSE is an excellent tool for identifying restenosis after PTCA. PMID- 8427171 TI - A shared pathway in atrioventricular nodal reentrant tachycardia and atrial flutter: implications for pathophysiology and therapy. AB - Atrioventricular (AV) nodal reentrant tachycardia and atrial flutter are considered 2 distinct supraventricular tachycardias. Recent clinical and experimental data suggest that both these tachycardias include an area in the lower right atrial septum in their reentrant pathways. This study was designed to test the hypothesis that there is an association between the mechanisms of AV nodal reentrant tachycardia and atrial flutter because of a shared pathway of reentry. Consecutive patients referred for evaluation and management of supraventricular tachycardia, thought to be due to AV nodal reentry, underwent electrophysiologic testing protocols designed to induce both AV nodal reentrant tachycardia and atrial flutter, if present. Fifteen of 29 patients (52%) had both AV nodal reentrant tachycardia and atrial flutter induced during electrophysiologic testing. Seven of these 15 patients (47%) underwent transcatheter radiofrequency current application (mean power 34 +/- 4 W) against the tricuspid annulus above the coronary sinus. In each patient, neither AV nodal reentrant tachycardia nor atrial flutter could be induced after the procedure. Repeat study after successful ablation (mean 6 days) showed no inducible supraventricular arrhythmia of either type at baseline study or during isoproterenol infusion. Atrial flutter occurs frequently (15 of 29 patients; 52%) in patients with AV nodal reentrant tachycardia, because of a shared pathway in their reentry circuits. Because of this shared pathway, both arrhythmias can be ablated at the same site. These observations promote new insights into the mechanism and therapeutics of supraventricular tachycardias. PMID- 8427172 TI - Immediate reproducibility of the tilt-table test in adults with unexplained syncope. AB - There are few data regarding the immediate reproducibility of the tilt-table test (TTT). Therefore, the immediate reproducibility of the TTT was examined in 19 patients (11 men and 8 women; mean age 49 +/- 19 years) with syncope or presyncope. The mean number of episodes that patients had experienced was 14 +/- 25 (range 1 to 100). After baseline supine observation for 10 minutes, patients were placed in 80 degrees of head-up tilt until a positive response occurred or for a maximum of 10 minutes. Patients were then returned to the supine position for 5 minutes, followed by retilt for another 10 minutes. If the baseline tilt was negative, the study was repeated with intravenous isoproterenol, and immediate reproducibility was examined in the same manner. The 19 patients underwent a total of 31 TTTs (19 baseline and 12 follow-up on drug). The TTT was immediately reproducible in 24 of 31 tests performed (77%). Eight tests were reproducibly positive and 16 negative. The results of 7 TTTs (23%) (5 baseline and 2 follow-up on drug) were not reproducible. In 6 of these studies (86%), the positive result occurred first and the negative result second. The reproducibility of an initially negative TTT result (16 of 17; 94%) was much higher than that of an initially positive one (8 of 14; 57%). The immediate reproducibility of the TTT in adult patients with unexplained syncope is approximately 75%. In studies that are not reproducible, most (86%) are positive first and negative second. Therefore, in most patients it is not necessary to check immediate reproducibility.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427173 TI - Congestive heart failure from left ventricular diastolic dysfunction in systemic hypertension. AB - Previous studies have pointed out that congestive heart failure (CHF) with normal ejection fraction presents a uniform clinical profile that is indistinguishable from heart failure with low ejection fraction. Thirty-six patients with systemic hypertension who had recently experienced CHF with normal ejection fraction (> or = 50%) and no clinical history of ischemic cardiomyopathy were studied. The patients were divided into 2 groups according to degree of echocardiographic hypertrophy: group A (19 patients) with a ventricular mass/volume ratio > 1.8, and group B (17 patients) with a ratio < 1.8. Group A patients had a higher ejection fraction (67 +/- 6 vs 57 +/- 3%, p < 0.01), smaller ventricular diameters and a lower thallium-201 positive rate at peak stress (10 vs 70% in group B, p < 0.001), with 8 of 10 showing severe coronary stenosis. Clinically, group A had a more frequent audible fourth sound (79 vs 17%, p < 0.001), a low incidence of audible third sound (5 vs 55%, p < 0.001) and a cardiothoracic ratio < or = 0.5 (63 vs 17%, p < 0.01). The degree of radionuclide-detected resting diastolic dysfunction and exercise intolerance was similar in both groups. In conclusion, CHF with normal ejection fraction in hypertensive patients presents 2 different profiles: one characterized by severe hypertrophy and the other by a high rate of myocardial regional ischemia. Therapy should be aimed at pathophysiologic regression of the hypertrophy in the first case, and at improvement of the ischemia in the second. PMID- 8427174 TI - Effects on left ventricular function of pindolol added to verapamil in hypertrophic cardiomyopathy. AB - The aim of the study was to assess the effect of adding pindolol to previously used verapamil monotherapy on left ventricular (LV) systolic and diastolic function in 20 patients with hypertrophic cardiomyopathy. Patients were initially treated with verapamil in maximal well-tolerated doses for 16 +/- 14 months; pindolol, 5 mg twice daily, was added. In a Doppler echocardiographic study all patients had altered LV diastolic filling despite verapamil therapy. The control examination, which consisted of echocardiographic study and New York Heart Association functional status classification, was performed after 20 days and repeated after > or = 6 months of follow-up. Combined pindolol and verapamil therapy caused an increase in LV diastolic filling manifested by beneficial changes of transmitral flow parameters. Also, inhibition of hypercontractile LV function expressed by reduction of LV outflow tract pressure gradient and ejection fraction was observed. New York Heart Association functional class was reduced in 13 patients. The magnitude and distribution of LV myocardial hypertrophy did not change significantly. It is concluded that pindolol and verapamil combined therapy is superior to verapamil monotherapy because of improved LV diastolic function (probably due to partial agonist activity of pindolol) and reduced hypercontractile function in patients with hypertrophic cardiomyopathy. PMID- 8427175 TI - Influence of clinical and hemodynamic characteristics on signal-averaged electrocardiogram in postoperative tetralogy of Fallot. AB - This study examines the relation between signal-averaged electrocardiographic measurements and the occurrence of spontaneous ventricular arrhythmias in 86 patients with a postoperative right bundle branch block after repair of tetralogy of Fallot; special attention was given to the influence of age, body surface area and right ventricular systolic pressure on signal-averaged electrocardiograms. Twenty-eight of the 86 patients had significant ventricular arrhythmias on 24 hour ambulatory monitoring. A positive linear correlation was found between filtered QRS duration and age at evaluation or body surface area (r = 0.45, p = 0.00001; r = 0.54, p < 0.00001, respectively) and between amplitude of the last 40 ms of the filtered QRS and right ventricular systolic pressure (r = 0.48, p < 0.001). A negative linear correlation was found between amplitude of the last 40 ms of the filtered QRS and age at evaluation or body surface area (r = -0.27, p = 0.01; r = -0.34, p = 0.002, respectively). When the age of the patients or the body surface area was considered with an analysis of covariance, the presence of ventricular arrhythmias was associated with a higher amplitude of the last 40 ms of the filtered QRS. In addition, an amplitude of the last 40 ms of the filtered QRS > 170 microV had an excellent sensitivity (100%) and a good specificity (88%) for identifying patients with both right ventricular systolic hypertension and spontaneous ventricular arrhythmia. Thus, adjustment of signal-averaged parameters for age at evaluation and body size is mandatory when studying postoperative tetralogy of Fallot. PMID- 8427176 TI - Rapidity of progression of aortic stenosis in patients with congenital bicuspid aortic valves. AB - The rapidity of progression of aortic stenosis in patients with congenital bicuspid aortic valves, and its relation to aging and valve anatomy are not well known. To elucidate these aspects, 75 patients aged 15 to 76 years were examined by echocardiography. Aortic valve sclerosis began from the second decade, the sclerotic index progressing with age (r = 0.72; p < 0.0001). Aortic valve calcium was noted from the fourth decade. Aortic valve pressure gradient increased approximately 18 mm Hg each decade, concomitant with progression of valve sclerosis (r = 0.78; p < 0.0001). Progression of cusp sclerosis was faster in patients with anteroposteriorly located cusps than in those with right-left located cusps (p < 0.005), and was faster in those with eccentric cusps (width ratio of major and minor cusps > or = 1.2) than in those with symmetric cusps (p < 0.05). In patients with eccentric and anteroposteriorly located cusps, aortic valve pressure gradient increased 27 mm Hg per decade. In patients with congenital bicuspid aortic valves, the progression of aortic stenosis is rapid, and the rapidity depends to some extent on the position and eccentricity of the cusps. PMID- 8427177 TI - Complications associated with percutaneous placement and use of intraaortic balloon counterpulsation. AB - In-hospital and late complications related to percutaneous placement of 240 intraaortic balloon pump catheters in 231 consecutive patients from March 1985 through June 1990 were reviewed. Mean age was 64 +/- 11 years and 34% were women. Average duration of counterpulsation was 44.2 hours. Indications for counterpulsation included complications of myocardial infarction (34.6%), prophylactic placement before high-risk coronary angioplasty (20.0%) or open heart surgery (12.9%), complicated coronary angioplasty (18.3%), end-stage cardiomyopathy (5.4%) and miscellaneous (8.8%). Early major complications occurred in 11 cases (4.6%) and included limb ischemia requiring surgery (n = 9), bleeding requiring arterial repair (n = 1) and septicemia (n = 1). Other complications included hematoma requiring transfusion (n = 7), limb ischemia resolving with balloon catheter removal (n = 12), and superficial wound infection (n = 1). Overall in-hospital complication rate was 13% (31 of 240). Peripheral vascular disease and diabetes were found to be significant predictors of limb ischemia (p = 0.01 and p = 0.02, respectively). Follow-up information was obtained in 97% of patients with a mean duration of 19 months: 2 patients (1.1%) required vascular surgery for femoral false aneurysms and 1 patient experienced new onset of claudication. In conclusion, compared with previous experience, contemporary intraaortic balloon counterpulsation with percutaneous placement of smaller size (8.5Fr to 10.5Fr) catheters is associated with improved complication profile. This will further enhance the current trend for an expanding role of intraaortic balloon counterpulsation in complex interventional procedures. PMID- 8427179 TI - Criteria for adoption in practice of medical practice guidelines. PMID- 8427178 TI - Persistence of coronary vasodilator responsivity after cardiac transplantation. AB - Accelerated graft atherosclerosis is a major cause of death after cardiac transplantation. Although its detection currently requires surveillance angiography, loss of vasodilator responsivity may precede obstructive lesions and be detectable by noninvasive assessment of myocardial perfusion. Thirty-five allograft recipients were studied an average of 31 +/- 19 (mean +/- SD) months after transplantation. All were free from angiographically definable macrovascular obstructive coronary artery lesions. Nutritive myocardial perfusion at rest, estimated in absolute terms by positron emission tomography with oxygen 15 water averaged 1.63 +/- 0.51 ml/g/min in patients and was greater than that in 26 healthy volunteers (1.17 +/- 0.33 ml/g/min, p < 0.001). The increase correlated with increased cardiac work at rest in transplant recipients with arterial hypertension and tachycardia. Peak myocardial perfusion induced by intravenous administration of dipyridamole was normal in the transplant recipients (3.49 +/- 1.70 ml/g/min compared with 3.60 +/- 1.41 ml/g/min in volunteers). Because of the high flow at rest, myocardial perfusion reserve (the ratio of hyperemic flow to flow at rest) was diminished (2.3 +/- 1.2 compared with 3.3 +/- 1.5 in volunteers, p < 0.005). These results indicate that the responsivity to vasodilator stimulation is well preserved in transplant recipients devoid of macroscopic coronary arterial lesions obviating detection of early vascular dysfunction in individual subjects. Positron emission tomography may be useful, however, in quantifying the magnitude of the increase in flow at rest secondary to increased cardiac work--a potentially remedial cause of accelerated coronary vascular disease induced by high shear force activation of platelets in the coronary bed, and in detecting impaired perfusion once macrovascular vascular disease is extant. PMID- 8427180 TI - Safety and efficacy of pentoxifylline in stable angina pectoris. PMID- 8427181 TI - Improved identification of late potentials by adjustment of the number of analyzed segments of the spectral temporal mapping of the signal-averaged electrocardiogram. PMID- 8427182 TI - Immediate normalization of profound sinus node dysfunction by aminophylline after cardiac transplantation. PMID- 8427183 TI - Optimal defibrillator patch configurations include the right side of the heart and left ventricle. PMID- 8427184 TI - Influence of systemic arterial blood pressure and nonhemodynamic factors on the brachial artery pulsatility index in mild to moderate essential hypertension. PMID- 8427185 TI - Initial outcome of percutaneous balloon valvuloplasty in rheumatic tricuspid valve stenosis. PMID- 8427186 TI - Element structure in stenotic mitral valves. PMID- 8427187 TI - Are hepatic pulsations in dilated cardiomyopathy with heart failure due to tricuspid regurgitation? PMID- 8427188 TI - Initial experience and potential applications of multiplane transesophageal echocardiography. PMID- 8427189 TI - Effects of nitroglycerin on platelet aggregation beyond the effects of acetylsalicylic acid in healthy subjects. PMID- 8427190 TI - Management of restenosis within the Palmaz-Schatz coronary stent (the U.S. multicenter experience. The U.S. Palmaz-Schatz Stent Investigators. PMID- 8427191 TI - Usefulness of transesophageal echocardiography in the diagnosis of ventricular septal rupture secondary to acute myocardial infarction. PMID- 8427192 TI - Sinus node deceleration during exercise as a marker of significant narrowing of the right coronary artery. PMID- 8427193 TI - Prospects for prevention of sudden coronary death. PMID- 8427194 TI - Effect of coronary stenosis severity on variability of quantitative angiography. PMID- 8427195 TI - Selenium deficiency, thyroid hormone metabolism, and thyroid hormone deiodinases. AB - Much research into the functions of selenium in the cell has concentrated on its role in selenium-containing glutathione peroxidases. However, selenium was recently shown to be an essential component of type I iodothyronine 5'-deiodinase in rats, which converts thyroxin to the more biologically active hormone 3,5,3' triiodothyronine. Thus, selenium-deficient rats have low tissue deiodinase activities and abnormal thyroid hormone metabolism. The discovery of this function for selenium in thyroid hormone metabolism has important implications for the interpretation of the effects of selenium deficiency, especially in individuals with an adequate vitamin E status. PMID- 8427196 TI - Effects of combined iodine and selenium deficiency on thyroid hormone metabolism in rats. AB - This paper compares the effects of combined iodine and selenium deficiency, of single deficiencies of these trace elements, and of no deficiency on thyroid hormone metabolism in rats. In rats deficient in both trace elements, thyroidal triiodothyronine (T3), thyroidal thyroxin (T4), thyroidal total iodine, hepatic T4, and plasma T4 were significantly lower, and plasma thyroid-stimulating hormone (TSH) and thyroid weight were significantly higher than in rats deficient in iodine alone. Plasma and hepatic T3 concentrations were similar in the dietary groups. Hepatic type I iodothyronine deiodinase (ID-I) activity was inhibited by selenium deficiency irrespective of the iodine status. Type II deiodinase (ID-II) activity in the brain was significantly higher and in pituitary, significantly lower in combined deficiency than in iodine deficiency alone. These data show that selenium can play an important role in determining the severity of the hypothyroidism associated with iodine deficiency. PMID- 8427197 TI - Selenium and the thyroid: how the relationship was established. AB - Several hypotheses concerning consequences of selenium deficiency on iodine metabolism can be proposed on the basis of experimental studies in rats and from epidemiological and experimental studies in humans. By decreasing intracellular GSH peroxidase activity, selenium deficiency may increase hydrogen peroxide (H2O2) supply and lead over several weeks to the thyroid atrophy observed in myxoedematous cretins. By improving thyroid hormone synthesis and by decreasing peripheral thyroxin (T4) deiodination, selenium deficiency could protect fetal brain T4 supply and thus prevent neurologic cretinism. Selenium deficiency may protect against iodine deficiency by decreasing T4 metabolism--and thus iodide leakage and--perhaps also by increasing H2O2 supply and thyroid hormone synthesis and thus thyroid efficiency. PMID- 8427198 TI - Molecular cloning of the selenocysteine-containing enzyme type I iodothyronine deiodinase. AB - It has proved impossible to purify the three types of iodothyronine deiodinase to homogeneity by use of standard physiochemical techniques. However, using expression cloning with Xenopus oocytes, we were successful in isolating a cDNA coding for the rat type I iodothyronine deiodinase. An extremely puzzling aspect of this cDNA was an in-frame thymine-guanine-adenine (TGA) codon at position 126. By mutagenesis studies we showed that this coded for selenocysteine, consistent with the previously described requirement for selenium for hepatic and renal thyronine deiodination. The cloned enzyme can be expressed transiently in JEG, a human cell line, or COS, a monkey cell line, by gene-transfer techniques, and the expressed protein has the appropriate biochemical characteristics of type I iodothyronine deiodinase. These include a newly identified extreme sensitivity to competitive inhibition of reverse-3,5,3'-triiodothyronine (rT3) deiodination by gold. The identification of this elusive protein should prove helpful in understanding the process of thyroid hormone activation. PMID- 8427199 TI - Indexes of selenium status in human populations. AB - This paper considers the factors that affect the bioavailability of selenium to human populations and describes briefly the consequences of an inadequate dietary intake of selenium in the Peoples' Republic of China and in Zaire. A review of human blood selenium concentrations worldwide reveals very large differences in the apparent dietary status of individuals in different areas. The question is raised as to whether blood selenium measurement is a reliable index of actual selenium status in terms of bioavailability and function of the element. It is concluded that the preferred indexes of human selenium status are blood, or plasma and/or serum, concentrations of the element and the level of activity of the selenium-dependent enzyme glutathione peroxidase in erythrocytes or in plasma. Several important caveats to this conclusion are mentioned and other means of assessing selenium status are also considered. PMID- 8427200 TI - The epidemiology of selenium deficiency in the etiological study of endemic diseases in China. AB - The distribution of chemical elements in rocks and soils varies widely. Selenium is one of the elements that has remarkable regional variations in distribution and bioavailability. This paper reviews the epidemiology of selenium deficiency in China in connection with the etiology of human selenium-responsive diseases, the well-defined Keshan disease (KD) and the less-well-defined Kashin-Beck disease (KBD). PMID- 8427201 TI - The epidemiology of iodine-deficiency diseases in China. AB - Iodine-deficiency diseases (IDDs) are wide-spread in China, distributed mainly in the inland and mountainous regions. About one-third of the total Chinese population lives in IDD-endemic areas. The severity of IDD is related to the severity of iodine deficiency. All the selenium-deficient areas of China are also IDD-endemic areas; however, IDD can be very severe in areas where selenium status is thought to be adequate. The distribution of myxedematous cretinism in China is not related to selenium deficiency. In the Tarim Basin, the selenium status of the population is normal and myxedematous cretinism is prevalent. In the northeastern regions of China, selenium deficiency is common and neurological cretinism is very rare. PMID- 8427202 TI - The epidemiology of iodine-deficiency disorders in relation to goitrogenic factors and thyroid-stimulating-hormone regulation. AB - In children aged 5-7 y from goiter-endemic areas in Ubangi, Zaire, and Ntcheu, Malawi, mean serum thyroxin (T4) concentrations were 53 +/- 49 vs 81 +/- 33 nmol/L (P < 0.05), and thyroid-stimulating hormone (TSH) values were 24.3 +/- 9.6 vs 4.5 +/- 3.3 mU/L respectively (P < 0.01); mean urinary iodine concentrations were 0.14 +/- 0.02 vs 0.09 +/- 0.02 mumol/L, and mean thiocyanate concentrations were 0.33 +/- 0.05 vs 0.17 +/- 0.05 nmol/L, respectively (P < 0.05). Mean serum selenium concentrations were 0.343 +/- 0.176 mumol/L in Ubangi and 0.437 +/- 0.178 mumol/L in Ntcheu (P < 0.05). In two groups of 11 adolescent girls from Ubangi, the mean values for excretion of urinary iodine were 1.31 +/- 0.14 and 0.58 +/- 0.17 mumol/L (P < 0.05) after a meal of cassava or a control meal of rice, respectively. In euthyroid subjects from Ubangi, mean serum TSH for a given serum T4 was approximately twice as high for children aged < 15 y than for those aged 16-25 y. The high frequency of myxedematous cretins observed in Ubangi very probably result from both severe iodine and selenium deficiency together with thiocyanate overload. PMID- 8427203 TI - Selenium deficiency mitigates hypothyroxinemia in iodine-deficient subjects. AB - Studies were performed to assess the role of combined selenium and iodine deficiency in the etiology of endemic myxedematous cretinism in a population in Zaire. One effect of selenium deficiency may be to lower glutathione peroxidase activity in the thyroid gland, thus allowing hydrogen peroxide produced during thyroid hormone synthesis to be cytotoxic. In selenium-and-iodine-deficient humans, selenium supplementation may aggravate hypothyroidism by stimulating thyroxin metabolism by the selenoenzyme type I iodothyronine 5'-deiodinase. Selenium supplementation is thus not indicated without iodine or thyroid hormone supplementation in cases of combined selenium and iodine deficiencies. PMID- 8427204 TI - Iodine-supplementation trials. AB - Iodine deficiency as a major determinant of endemic goiter has been recognized for several decades. The evidence is also strong that iodine supplementation is effective in preventing both varieties of endemic cretinism, provided it is given before conception; early fetal and infant death may also be prevented. However, the reason for the variation in prevalence of the different types of cretinism is still a matter of conjecture. Iodine supplementation may improve motor and cognitive performance, but the evidence needs to be evaluated in light of the difficulty in transposing tests of intellectual and motor ability developed in one culture to another very different one. PMID- 8427205 TI - Effects of iodine deficiency on thyroid hormone metabolism and the brain in fetal rats: the role of the maternal transfer of thyroxin. AB - Thyroid hormones, thyroxin (T4) and 3,5,3'-triiodothyronine (T3), of maternal origin, are available to the mammalian embryo early in development. However, after the onset of fetal thyroid function, they are of both fetal and maternal origin. Maternal T4 has a protective effect on the fetal brain in cases of congenital hypothyroidism. In severe iodine deficiency, maternal T4 is low, although T3 is normal; the developing embryo is markedly T4-deficient; and T3 deficiency increases with gestational age. In contrast to mechanisms in the hypothyroid fetus from a normal mother, the low T4 of the iodine-deficient mother prevents any protective effects on the fetal brain. Thyroid hormone deficiency of the iodine-deficient fetus, including the brain, is more severe and prolonged than it is in the cases of maternal or fetal thyroid failures. These findings may help to explain the relationship between severe maternal hypothyroxinemia and the severe central nervous system damage of the neurological endemic cretin. PMID- 8427206 TI - Effects of nutrition on brain development in humans. AB - Brain development in humans is remarkably resistant to permanent damage from protein-energy malnutrition. However, specific nutrients have crucial roles. Iodine deficiency is the most important and widespread nutrient deficiency; it causes endemic cretinism, associated with deaf-mutism and cerebral palsy. Iodine deficiency during pregnancy causes both maternal and fetal hypothyroxinemia, resulting in irreversible impairment of brain development at a critical stage. Neuropathological data place this after 14 wk, perhaps continuing through the third trimester. Gross brain structure, including the gyral pattern of the cerebral cortex, develops normally; the insult affects neuron and dendrite growth. Recent magnetic-resonance-imaging (MRI) images of neurological cretin brains show remarkably normal appearance except for gliotic lesions of the globus pallidus, correlating with the proximal motor rigidity seen clinically. Myxedematous cretinism is paradoxical in showing more severe hypothyroidism and growth failure, yet better intellectual, motor, and hearing function; these observations implicate a second independent factor in its pathogenesis. PMID- 8427207 TI - 5'-Monodeiodinase activity in developing human cerebral cortex. AB - Iodine status of aborted fetuses (11-25 wk gestation) was determined on the basis of maternal thyroid status and urinary iodine excretion. Fetal cerebral cortex thyroxin and 3,5,3'-triiodothyronine (T3) contents peaked at 15-18 wk gestation and then fell in iodine sufficiency. In mild iodine deficiency T3 concentration was maintained at a higher level until week 22, although it were still significantly less than in the iodine-sufficient group. Reverse T3 (rT3) rose from 11 to 22 wk gestation without any effect of iodine status. The activity of 5' and 5-monodeiodinase also increased from 11 to 22 wk gestation, with 5' monodeiodinase activity significantly increased by mild and moderate iodine deficiency. In contrast, cerebral cortex 5-monodeiodinase activity was significantly lower in moderate iodine deficiency at 15-18 wk gestation. The observations indicate that cerebral cortex 5' and 5 monodeiodinase activities are modulated in iodine deficiency to enhance T3 production from thyroxin (T4) during the period that coincides with neuroblastogenesis. PMID- 8427208 TI - Testing the effects of nutrient deficiencies on behavioral performance. AB - The association between specific nutrient deficiencies and poor performance on behavioral tests has been documented for several nutrients. The determination of causality, however, remains elusive. This paper presents the essential criteria for a valid test of causality. Findings from experimental studies in which a nutritional treatment was randomly allocated can be summarized in a statistical statement about the probability that the nutrient treatment caused the behavioral response. Criteria for assessing the internal validity of these studies are examined in terms of whether alleviation of a nutrient deficiency did or did not produce a detectable behavioral response. The plausibility of such a causal inference is dependent on its congruency with known or theorized biological and behavioral mechanisms. External validity describes the extent to which inferences from internally valid studies may be applicable to other populations or circumstances. In addition to these scientific considerations, some of the ethical issues of nutrient-treatment trials are also discussed. All of these considerations provide a better basis for judging whether public health action would be worthwhile than do observed associations that could actually be due to other causes. PMID- 8427209 TI - Assessments of the effects of nutrition on mental development and behavior in Jamaican studies. AB - Methodological issues and problems in studies of the effects of protein-energy malnutrition (PEM) and child development are discussed with reference to our experience in Jamaica. The usefulness of defining PEM by stunting, wasting, and edema is stressed. Difficulties in achieving reliable and valid measurements of mental development and behavior in Third World children, and allowing for the confounding and possibly interacting effects of social background, are discussed. Problems associated with different approaches are outlined. PMID- 8427210 TI - Effects of dietary selenium on the tissue concentrations of type I iodothyronine 5'-deiodinase and other selenoproteins. AB - Since selenium was identified as the essential component of the type I iodothyronine 5'-deiodinase, it has become important to determine the selenium dose necessary for the formation of the deiodinase. In this survey the results of studies of the effects of dosage and chemical form of the element on the concentrations of this enzyme and of other selenoproteins are discussed. They show that the selenium requirement for normal deiodinase activity is lower than it is for adequate glutathione peroxidase activity. Elevated tissue selenium concentrations found after intake of higher doses of the element, especially when given in the form of selenomethionine, are due to nonspecific incorporation into other proteins. The tissue concentrations of the selenonenzymes appear to be homeostatically controlled and cannot be further increased by additional selenium supplementation. PMID- 8427211 TI - Selenium-dependent regulation of type I 5'-deiodinase expression. AB - Selenite concentration regulates activity and expression of the p27 substrate binding subunit of type I 5'deiodinase (5'-D) and of a protein labeled with bromoacetylthyroxin (BrAcT4), or p30, with yet unknown function in a porcine kidney epithelial cell line (LLC-PK1) cultured in serum-free medium. p27 is metabolically labeled by 75-selenite and affinity labeled by BrAc[125I]T4. Compared with glutathione peroxidase, expression of the p27 5'D subunit (5'-DI) is observed at 10-fold-lower concentrations of selenium in the growth medium, suggesting an intracellular hierarchy of selenite utilization. Selenium deficiency retards cell growth and prevents 5'-DI expression and may thus impair thyroid hormone action in vivo. PMID- 8427212 TI - Goiter problems in Ethiopia. AB - Survey studies have shown that there is wide-spread goiter in Ethiopia (mean incidence 25%). The occurrence of goiter is associated with low iodine intakes, but little information has been available on goitrogens. A program of distribution of iodized salt has been initiated to eliminate the goiter problem. PMID- 8427213 TI - Key issues in generating a psychological-testing protocol. AB - An individual's behavior is created by an amalgam of genetic, environmental, cultural, and historical variables working in concert and changing over time. Variations in nutrition are one class of environmental factors. Linking these to outcome effects requires carefully designed studies. Many considerations are involved, and this paper draws attention to some of the fundamental ones. Psychological and behavioral functions tend to be complex, in part because they are affected by a wide range of variables. Almost any given state--a particular constellation of psychological characteristics displayed by an individual at a given time--can be produced in a variety of ways. PMID- 8427214 TI - An American Board of Orthodontics case report: the nonsurgical orthodontic correction of a Class III malocclusion. AB - A case report about the orthodontic treatment of a Class III malocclusion in a black man 22 years 8 months old. The patient's chief complaint concerned the space left by the traumatic loss of the maxillary left central incisor. Not desiring changes in facial appearance, the patient sought an orthodontic solution to his problem. An acceptable result was achieved through maxillary expansion and the reduction of mandibular space. [This case was presented to the American Board of Orthodontics in partial fulfillment of the requirements for the certification process conducted by the Board.] PMID- 8427215 TI - Biodegradation of orthodontic appliances. Part II. Changes in the blood level of nickel. AB - The purpose of this study is to determine whether orthodontic patients accumulate measurable concentrations of nickel in their blood during their initial course of orthodontic therapy. Blood samples were collected at three different time periods: before the placement of orthodontic appliances, 2 months after their placement, and 4 to 5 months after their placement. The study involved 31 subjects, 18 females and 13 males, who had malocclusions that required the use of a fully banded and bonded edgewise appliance. The age of the subjects in the study ranged between 12 and 38 years. The blood samples were frozen and shipped to a commercial medical laboratory for analysis by atomic absorption spectrophotometry. The three blood samples for each patient were analyzed in succession on the same day to eliminate equipment variance that could occur if blood samples were analyzed on separate days. A total of 93 blood samples were sent for analysis. From the findings in this study the following can be concluded: (1) Patients with fully banded and bonded orthodontic appliances did not show either a significant or consistent increase in nickel blood levels during the first 4 to 5 months of orthodontic therapy. (2) Orthodontic therapy using appliances made of alloys containing nickel-titanium did not result in a significant or consistent increase in the blood levels of nickel. The results obtained from both parts of this investigation indicate that orthodontic appliances used, in their "as-received" condition, corrode in the oral environment releasing both nickel and chromium, in amounts significantly below the average dietary intake.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427216 TI - Repair process of external root resorption subsequent to palatal expansion treatment. AB - The repair process of external root resorption (ERR) and the role of retention mechanics in enhancing ERR repair were studied on eight Macaca fascicularis monkeys that were divided equally into short- and long-term groups. Six monkeys received palatal expansion appliances, and two received sham appliances. The short-term group received active treatment. The long-term group received additional retention (4 months) and relapse (2 months) treatment periods with biweekly injections of individual vital dye per phase, i.e., procion red H-8B and violet H-3R (80 mg/kg B.W.), respectively. Histomorphologic examinations included scanning electron microscopy, light, and ultraviolet microscopy. The short-term group demonstrated penetrated resorption with pulp exposure at sites with initial deficiency of the protecting odontoblastic layer (apical zone, nutrition canal). The long-term group showed two forms of ERR repair: (1) Nonfunctional retarded repair cementum, comprised of overlapped incremental lines and deprived of extrinsic fibers, was delineated in severe pulp exposure. The pulp/dentin complex showed intense incorporation of procion dye in the dentinal tubuli, conceivably related to a defense response in the form of sclerotic dentin. (2) Functional rapid repair cementum, comprised of discriminated incremental lines mainly of mixed cellular cementum, with a consistent pattern of five sequential phases: the lag phase (14 to 28 days), the incipient phase (14 days), the peak phase (14 to 28 days), the steady phase (42 to 56 days) and the retreating phase (70 days). Sharpey's fibers at functional ERR sites were scarce, never emerging from the dentinocemental junction, and not developing into principal fibers. The pulp/dentin complex showed an increase in pulp stones but no formation of tertiary dentin. The apical area responded by hypercementosis in the form of apical occlusion and a displaced pulp canal. The application of a fixed retention device is recommended in light of accelerated apposition of repair cementum during the retention period. However, increased formation of Sharpey's fibers during the relapse period might suggest a restricted duration in splinting therapy. PMID- 8427217 TI - Treatment of identical twins with Frankel and Herbst appliances: a comparison of results. PMID- 8427218 TI - Root resorption after orthodontic treatment: Part 2. Literature review. AB - All permanent teeth may show microscopic amounts of root resorption that are clinically insignificant and radiographically undetected. Root resorption of permanent teeth is a probable consequence of orthodontic treatment and active tooth movement. The incidence of reported root resorption during orthodontic treatment varies widely among investigators. Usually, extensive resorption does not affect the functional capacity or the effective life of the tooth. Most studies agree that the root resorption process ceases once the active treatment is terminated. Root resorption of the deciduous dentition is a normal, essential, and physiologic process. Permanent teeth have the potential to clinically undergo significant external root resorption when affected by several stimuli. This resorptive potential varies in persons and between different teeth in the same person. This throws doubt on the role of systemic factors as a primary cause of root resorption during orthodontic treatment. Tooth structure, alveolar bone structure at various locations, and types of movement may explain these variations. The extent of treatment duration and mechanical factors definitely influence root resorption. In most root resorption studies, it is not possible to compare the results and conclusions because of their different methods. Further research in this field is necessary to advance the service of the specialty. The question of whether there is an optimal force to move teeth without resorption or whether root resorption may be predictable remain unanswered. This review indicates the unpredictability and widespread incidence of the root resorption phenomenon.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427219 TI - Change in lip vermilion height during orthodontic treatment. AB - The extent that the height of the vertical aspect of the vermilion of the lips decreases during treatment may determine the ultimate esthetic results for a particular patient. Marked decrease in vertical height of the vermilion may prove to be esthetically pleasing for the patient with excessive vermilion; whereas it may prove esthetically disastrous for the patient exhibiting relatively small vermilion heights before treatment. Frontal esthetics should equal the profile in importance when treatment planning. The objectives of this study were (1) to quantify vermilion height changes when incisors are retracted, (2) to determine whether the pretreatment vertical position of the upper lip on the maxillary central incisor is associated with vermilion height changes, and (3) to relate vermilion height changes to incisor retraction. Cephalometric films from 40 adult female orthodontic patients (20 Class I and 20 Class II, Division 1) were measured with dial calipers. Significant decrease (paired t tests, P < 0.05) of the mean vermilion heights of both lips occurred during treatment in the patients with Class I relationships (upper 0.75 mm, lower 0.95 mm) and Class II, Division 1 relationships (upper 0.75 mm, lower 0.60 mm).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427220 TI - Laser debonding of ceramic orthodontic brackets. AB - Laser light energy has been shown in other studies to degrade resins by thermal softening, thermal ablation, or photoablation. If this technology could be successfully applied to bracket debonding, fracturing of both bracket and enamel during debonding might be eliminated. Both polycrystalline alumina and single crystal alumina (sapphire) ceramic orthodontic brackets were bonded to the labial surfaces of lower deciduous bovine incisor teeth with the acid-etch technique as currently practiced in dentistry. Under an externally applied stress of either zero or 0.8 MPa, the brackets were debonded by irradiating the labial surfaces of the brackets with laser light at wavelengths of 248 nm, 308 nm, and 1060 nm, and at light power densities of between about 3 and 33 W/cm2. Debonding times were measured, and the surfaces created by debonding were examined with both light and scanning electron microscopy to determine the extent of bracket and enamel damage. The results showed that under the conditions of this study, no enamel or bracket damage was present in any sample. The polycrystalline brackets debonding times were about 3 seconds, 5 seconds, and 24 seconds for 248 nm, 308 nm, and 1060 nm of radiation, respectively. The debonding of polycrystalline brackets is caused by thermal softening of the bonding resin resulting from heating of the bracket. The hot bracket then slides off the tooth. All sapphire brackets debonded in less than 1 second. At sufficiently high power levels, debonding of sapphire brackets is caused by either thermal ablation or photoablation resulting from direct interaction of the light beam with the resin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427221 TI - Contemporary trends in orthodontic practice: a national survey. AB - This study involved the development of a questionnaire to elicit information about current trends in orthodontic practice compared with trends of 5 years ago. The subjects addressed included the use of fixed appliances, functional appliances, extraction therapy, orthodontics and the temporomandibular joint, current diagnostic aids, and medicolegal implications of orthodontic treatment. The questionnaire was mailed to 1400 orthodontic specialists who were randomly selected from across the United States. There were 814 questionnaires returned, representing a 58.14% response rate. The major results of this survey are as follows: (1) The reported extraction rate has declined from a mean of 37.74% 5 years ago to 29.28% today. (2) TMJ concerns (or medicolegal implications thereof) have had a considerable impact in this decline, with 26.4% of orthodontists being influenced, to some extent, to extract fewer teeth because of this factor. (3) The use of functional appliances has remained somewhat static over the last 5 years, after a period of rapid growth in the 5 years previous to that. (4) The preadjusted edgewise appliance is by far the most popular fixed appliance in use today, being chosen by 72.6% of respondents. Analysis of the overall results lead to the following conclusions: (1) Fixed appliance therapy is the therapy of choice of the overwhelming majority of orthodontists. The use, benefits, and role in orthodontics of functional appliance therapy is considerably less defined. (2) Treatment modalities, notably on the use of extraction and its relationship to the health of the temporomandibular joint, should be determined by research, not by legal fears or unsubstantiated allegation. PMID- 8427222 TI - Safer orthodontic debonding with rubber dam. PMID- 8427223 TI - Comments on debonding of brackets. PMID- 8427224 TI - Comments on debonding of brackets. PMID- 8427225 TI - Recycled brackets. PMID- 8427226 TI - Nonextraction treatment of a Class II, division 1 malocclusion. PMID- 8427227 TI - Adoption of a new Haemophilus influenzae type b vaccine recommendation. PMID- 8427229 TI - The diagnosis of child sexual abuse. PMID- 8427228 TI - MMR vaccine and neomycin allergy. PMID- 8427230 TI - Current opinion: indications for initiating dexamethasone therapy in children with meningitis in the emergency department. PMID- 8427231 TI - Unrecognized exercise-induced bronchospasm in adolescent athletes. PMID- 8427232 TI - Age and sex as risk factors for burn deaths among children. PMID- 8427233 TI - Should we perform lumbar punctures on the first day of life? PMID- 8427234 TI - Acute mountain sickness in children at 2835 meters. AB - OBJECTIVE: Acute mountain sickness has been described in adults but little is known concerning its occurrence in children. Our objective was to determine the incidence of acute mountain sickness in children. METHODS: A survey questionnaire was completed by 558 children (aged 9 to 14 years) after they ascended from 1600 to 2835 m and from 405 similarly aged children after travel at sea level. RESULTS: Three or more of the following symptoms in the high-altitude setting were considered as the case definition of acute mountain sickness: headache, loss of appetite, vomiting, fatigue, insomnia, shortness of breath, and dizziness. One hundred fifty-six (28%) of the children at 2835 m developed acute mountain sickness. Three or more symptoms developed in a smaller, but nonetheless considerable, number (86 [21%]) of children at sea level. Headache, shortness of breath, and dizziness were reported more frequently at high altitude than at low altitude, whereas the other symptoms occurred with equal frequency at the two locations. CONCLUSIONS: More than one fourth of the children visiting high altitude developed acute mountain sickness. A high proportion (21%) of children at sea level developed similar symptoms, suggesting that an appreciable portion of the symptoms present were due to factors other than altitude, such as travel, anxiety, or disruption of daily routine. PMID- 8427235 TI - Physicians treating their own children. AB - OBJECTIVES: To determine the types of health care interventions physicians provided to their own children, identify those conditions most often treated by physician-parents, compare the differences in treatment practices among physician groups, and explore the reasons physicians give for treating their own children. DESIGN: Questionnaire. SETTING: University-affiliated teaching hospital in Iowa City, Iowa. PARTICIPANTS: Three hundred fifty-three physicians in residency and fellowship training and faculty who were parents of children aged 18 years or younger. INTERVENTIONS: None. MEASUREMENTS/MAIN RESULTS: The majority of physicians reported treating their afebrile child for acute illness. Fifty-five percent of physicians reported that they rarely or never treated their febrile child (temperature > 38.3 degrees C) without consultation with the child's physician. Only 47% of physicians reported that they always performed a physical examination on their child before treating. Physician-parents were more likely to auscultate the child's chest or perform otoscopy and less likely to obtain urine samples or throat swabs for culture before treating. Sixty-two percent of physicians reported that they have never performed routine health maintenance examinations on their own children, but 29% referred their children to a specialist. Medications were prescribed for their children by 65% of physicians. Neither gender nor level of training influenced the treatment practices of physician-parents. Primary care physicians were less likely to contact the child's physician for advice in treating their febrile child than were subspecialists. Pediatricians more often treated their afebrile and febrile children, performed physical examinations and laboratory studies, and prescribed medications than did other primary care physicians. Convenience was the most important reason physicians gave for treating their own children. CONCLUSIONS: Physicians frequently treat their own ill children, prescribe medications for them, and self-refer them to specialists largely for the sake of convenience. PMID- 8427236 TI - The children's vaccine initiative. PMID- 8427237 TI - High prevalence of overweight in inner-city schoolchildren. AB - The weights and heights of 1670 Harlem children aged 5 to 11 years (879 boys, 791 girls) were compared with the National Center for Health Statistics standards. The median height for age and weight for age were greater by 1.4 cm and 2.2 kg in boys and 2.7 cm and 2.8 kg in girls. Weight distribution was skewed to the right and showed a bimodal pattern. "Height-ages" were advanced by an average of 0.24 years in boys and 0.47 years in girls. Weight for height in 13.9% of girls and 13.6% of boys were above the 95th percentile. The results are consistent with early maturation. However, even after adjusting for height, shift to the right of the weight distribution persists with a large subgroup of boys and girls who are overweight. PMID- 8427238 TI - Comparison of two hypocaloric diets in obese children. AB - OBJECTIVE: To examine whether a protein-sparing modified fast diet and a hypocaloric balanced diet are safe and effective for children in an outpatient weight reduction program. DESIGN: Randomization of two groups to either diet, with follow-up at 14.5 months. SETTING: Physician or parent referral to outpatient program at Children's Hospital of New Orleans, La. PARTICIPANTS: Nineteen children, ranging in age from 7.5 to 16.9 years, weighing 45% to 131% more than the mean weight for age, sex, and height. INTERVENTION: During the first 10 weeks, 10 children were placed on a protein-sparing modified fast diet (2520 to 3360 J), and nine children and adolescents were placed on a hypocaloric balanced diet (3360 to 4200 J). Subsequently, all participants were placed on a hypocaloric diet; calories were increased from 4200 to 5040 J in a 3-month period and maintained for 1 year. SELECTION PROCEDURES: Children were assigned to one of two diets for the first 10 weeks according to their time of enrollment. MEASUREMENTS/MAIN RESULTS: Both diets produced significant weight loss during the first 6 months. However, the protein-sparing modified fast diet produced significantly greater changes in the percentage of overweight at 10 weeks (-30% vs -14%) and at 6 months (-32% vs -18%). At 10 weeks, a significant loss of adipose tissue with preservation of lean body mass occurred in the protein sparing modified fast group. A transient slowing of growth velocity was noted at 6 months in both dietary groups compared with values at 14.5 months. Growth velocity approached normal levels at 14.5 months compared with standards for North American children. When dietary groups were combined, the initial mean blood pressure decreased significantly at all points in the study. The initial mean serum cholesterol value also decreased significantly at 10 weeks. No biochemical or clinical complications were observed. CONCLUSIONS: These hypocaloric diets appear to be safe and effective in the short-term management of pediatric obesity. However, these diets should not be used without close medical supervision. PMID- 8427239 TI - Cerebral salt wasting in children. The need for recognition and treatment. AB - OBJECTIVES: To describe a salt-wasting syndrome in children with central nervous system (CNS) insults and to differentiate it from the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) and diabetes insipidus so that it may be more readily diagnosed and treated. DESIGN: Case reports. SETTING: Community teaching hospital. PATIENTS: Two inpatients with CNS insults (closed head trauma in one and seizure disorder, spastic diplegia, mental retardation, and hydrocephalus in the other). SELECTION CRITERIA: Evidence of hyponatremia accompanied by elevated urine sodium concentration and excessive urine output. INTERVENTIONS: Volume-for-volume urine replacement with 0.9% and/or 3% sodium chloride. Oral salt supplementation was required for brief periods to maintain normal plasma sodium concentration after discharge from the hospital. MEASUREMENTS AND MAIN RESULTS: Both patients had hyponatremia, high urine sodium concentrations, hypovolemia, and excessive urine output while receiving maintenance fluids. They also had elevated plasma atrial natriuretic hormone (ANH) concentrations, decreased aldosterone concentrations, and decreased [corrected] plasma renin activity for their degree of hyponatremia and negative fluid balance. Both patients maintained normal serum electrolyte concentrations with appropriate treatment. CONCLUSIONS: These patients showed true salt wasting associated with acute or chronic CNS injury, with hormonal patterns consistent with "inappropriate" ANH secretion and distinct from the SIADH. It is important to distinguish cerebral salt wasting (CSW) from the two other major disturbances of water metabolism seen following CNS injury (ie, SIADH and diabetes insipidus), because incorrect diagnosis and treatment could greatly increase morbidity in CSW. The etiologic roles of ANH or brain natriuretic peptide in CSW need to be further elucidated. PMID- 8427240 TI - Familial isolated growth-hormone deficiency with advanced sexual maturation. AB - Two brothers, aged 15 1/2 and 13 1/2 years, with dwarfism, microcephaly, and advanced sexual and skeletal maturation are described. One patient was mentally retarded. The parents were first cousins. Endocrine studies of these patients documented low growth-hormone levels after clonidine and insulin stimulation and blunted growth-hormone response to growth hormone releasing hormone. Gonadotropin releasing hormone stimulation produced no changes in levels of luteinizing and follicle-stimulating hormones. Basal levels of 17-alpha-hydroxyprogesterone were elevated in the two patients and increased further in response to stimulation with corticotropin. Levels of testosterone, dehydroepiandrosterone sulfate, and androstenedione were variably increased in both patients and showed a proportional increase on stimulation with human chorionic gonadotropin. To our knowledge, this is the first report of a familial association between growth hormone deficiency and advanced bone and sexual maturation. A pituitary and an independent adrenal defect could account for the observations in these patients, but in view of the familial recurrence, a common underlying defect is possible. PMID- 8427241 TI - Preparing residents for Public Law 99-457. A survey of pediatric training programs. AB - Recent implementation of Public Law 99-457 has high-lighted the integral role of pediatricians in early intervention for infants and young children (birth to age 3 years) with disabilities and their families. Faculty representatives of all US pediatric residency programs were surveyed to determine the current status of resident education about issues in this content area. Of 219 surveys, 159 (73%) were returned and analyzed. Resident rotations in child development were offered in 89% of the programs. Of these, 73% were mandatory rotations. Generally, comprehensive resident exposure to a majority of important topics regarding Public Law 99-457 was reported. However, minimal teaching by parents and community agencies was used. Content areas perceived to be strengths in most programs included normal and atypical infant development and developmental assessment. Weaker areas included cultural aspects of family function, case management, and advocacy. Barriers to optimal teaching included time constraints of residents and faculty, inadequate resources and facilities, and insufficient resident interest. Respondents expressed strong interest in improving resident education in this area. PMID- 8427242 TI - The use of primidone in neonates with theophylline-resistant apnea. AB - OBJECTIVE: To determine whether primidone reduced the occurrence of apnea of prematurity in neonates with apnea resistant to theophylline. DESIGN: Retrospective review. SETTING: Neonatal intensive care unit. PARTICIPANTS: Sixteen premature infants (mean age, 27.8 weeks) in whom apnea and bradycardia recurred despite therapeutic levels of theophylline. Six of the patients were receiving assisted ventilation. INTERVENTION: Administration of primidone (10 to 15 mg/kg per day) orally or by nasogastric tube at a mean age of 35 days. RESULTS: Apnea and bradycardia decreased significantly 24 to 72 hours after initiation of primidone treatment (by 68% and 69%, respectively) compared with pretreatment events. We obtained similar results after a separate analysis of the 10 patients who had been weaned from assisted ventilation before treatment with primidone. No toxic reactions were observed. CONCLUSIONS: Primidone represents a possible adjuvant therapy in theophylline-resistant apnea of prematurity. Caution is advised, because of primidone's complex pharmacologic characteristics, until there are further controlled prospective studies. PMID- 8427243 TI - The effect of antenatal dexamethasone administration on the fetal and neonatal ductus arteriosus. A randomized double-blind study. AB - OBJECTIVE: To determine whether antenatal dexamethasone sodium phosphate administration constricts the fetal ductus arteriosus or improves spontaneous closure of the ductus in premature infants. DESIGN: A randomized double-blind study. SETTING: University hospital of Helsinki, Finland. PARTICIPANTS: Sixty-one pregnant women with threatened preterm delivery between 24 and 31.9 weeks' gestation and 57 of their offspring (28 in the dexamethasone and 29 in the placebo group), born at 24 to 34.9 weeks' gestation, were studied using Doppler echocardiography. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Between the dexamethasone and placebo groups, there were no significant differences in systolic or diastolic flow velocity values in the fetal ductus arteriosus. Of the 29 infants with respiratory distress syndrome, 26 (90%) had hemodynamically significant patent ductus arteriosus and received indomethacin sodium: 12 (92%) of 13 dexamethasone-treated infants and 14 (88%) of 16 placebo-treated infants. One placebo-treated infant was ligated. Of the 28 infants without respiratory distress syndrome, only four (14%) had hemodynamically significant patent ductus arteriosus. In infants born at or before 30 weeks' gestation, spontaneous closure of the ductus occurred more frequently after administration of prenatal dexamethasone (in six of 17 infants; 35%) than placebo (in none of 10 infants; P < .05). CONCLUSIONS: Our data indicate that antenatal dexamethasone administration had no constrictive effect on the ductus arteriosus of the fetus between 24 and 31 weeks' gestation. However, antenatal dexamethasone had a beneficial effect on ductal closure in very premature infants. PMID- 8427244 TI - Mesenchymal hamartoma of liver. A regional ischemic lesion of a sequestered lobe. AB - OBJECTIVE: To evaluate histologic and gross features of mesenchymal hamartoma of liver and similar lesions in relation to determining likely pathogenesis. DESIGN: Case series of patients presenting to a tertiary care hospital over 16 years. PATIENTS: Three children with mesenchymal hamartoma ranging from newborn to 11 months of age and one 12-year-old girl with torsion of an accessory lobe of liver. MAIN OUTCOME MEASURES: Similarity of gross vascular and segmented anomalies as well as apparent consequent histologic features of mesenchymal hamartoma to those of torsion of an accessory lobe of liver indicate that they are pathogenetically related. RESULTS: Dissecting microscopic examination revealed a single vascular supply in one case and remote thrombi in two cases of mesenchymal hamartoma evaluated. The histologic features of mesenchymal hamartoma (hypocellular central zone and hypercellular periphery) were duplicated in the lobe of liver with torsion. CONCLUSIONS: Mesenchymal hamartoma represents a lesion with an anomalous solitary vascular supply that may evolve into its specific pattern with stromal cysts as a result of early ischemic changes. PMID- 8427245 TI - Eosinophilic pustular folliculitis in infancy and childhood. AB - We describe nine patients with eosinophilic pustular folliculitis occurring in infancy. All patients had recurrent crops of pruritic papules primarily affecting the scalp and brow region of the face. Biopsy specimens revealed folliculitis with a predominant eosinophilic infiltrate. Bacterial and fungal cultures of pustules were negative. Most patients had leukocytosis and eosinophilia. Clinical response to topical corticosteroid and/or systemic antibiotic therapy was noted. Eosinophilic pustular folliculitis of infancy must be considered in the differential diagnosis of pustular disorders in infancy. PMID- 8427246 TI - Radiological cases of the month. Presentation of lymphoma with renal insufficiency. PMID- 8427247 TI - Radiological case of the month. Odontogenic keratocyst of the maxillary sinus. PMID- 8427248 TI - Picture of the month. Campomelic dysplasia. PMID- 8427249 TI - Pathological case of the month. Junctional epidermolysis bullosa. PMID- 8427250 TI - Pathological case of the month. Juvenile granulosa cell tumor. PMID- 8427251 TI - Mortality and the acquisition of basic skills by children and adults with severe disabilities. AB - OBJECTIVE: To determine normative data on age-related probabilities of children with severe disabilities acquiring mobility or self-feeding skills, or dying during a 5-year follow-up period. RESEARCH DESIGN: A 5-year follow-up study of three mutually exclusive subgroups formed on the basis of severe, profound, or suspected levels of retardation and incontinence and the following combinations of feeding and mobility skills. PARTICIPANTS: The sample was made up of 7836 children and adults distributed among the three subgroups being served in California between January 1981 and December 1985. MEASUREMENTS/MAIN RESULTS: Subjects who were tube-fed and immobile showed very little likelihood of becoming mobile or feeding themselves and had a high probability of death. Individuals who had some mobility experienced a better outcome. CONCLUSIONS: After age 6 years, the most probable outcome for children who are immobile and cannot feed themselves is death or no improvement in self-help skills. PMID- 8427252 TI - Uses of nonsteroidal anti-inflammatory drugs in pediatrics. AB - Nonsteroidal anti-inflammatory drugs have an expanding role in pediatrics. The indications include (1) treatment of childhood rheumatic disease and other chronic inflammatory conditions; (2) symptomatic treatment of clinical problems such as fever, musculoskeletal pain, or dysmenorrhea; and (3) induction of closure of patent ductus arteriosus. Nonsteroidal anti-inflammatory drugs are generally well tolerated with a low incidence of serious side effects. However, potentially serious gastrointestinal, hepatic, central nervous system, and renal adverse effects have occurred. The pediatrician should be familiar with the pharmacokinetics and dosages of the various nonsteroidal anti-inflammatory drugs as well as laboratory tests necessary to monitor their use. Guidelines are suggested. PMID- 8427253 TI - Current methods of estimating severity for occupational injuries and illnesses: data from the 1986 Michigan Comprehensive Compensable Injury and Illness Database. AB - National and state estimates of the severity of occupational injuries and illnesses (severity = lost work time = missed work days+restricted work days) have come from the annual Survey of Occupational Injuries and Illnesses (Survey) produced by the U.S. Bureau of Labor Statistics. However, we show that the Survey practice of collecting injury information soon after the accident year reduces substantially the accuracy of missed work day estimates, which constitute 85.3% of the Survey lost work time estimate. To develop an independent estimate of missed work days, the research team created the Michigan Comprehensive Compensable Occupational Injury Database (Michigan Database) by linking state files with injury characteristics to files with workers' compensation information for injuries occurring in 1986. The measure of missed work time (days, weeks, or years) is the cumulative duration of compensation from the "date disability commenced," noted on the first payment form, through follow-up to March 1, 1990. Cumulative missed work time has been calculated or estimated for 72,057 injured workers, more than 97% of the 73,609 Michigan workers with compensable occupational injuries in 1986 identified through the close of the study. Our "best" estimate of missed work days, to follow-up, attributable to both fatal and nonfatal compensable occupational injuries and illnesses is 7,518,784, a figure four times that reported for Michigan by the Survey. When insurance industry data on disbursements are also considered, the estimate of missed work days increases to 8,919,079, a figure 4.75 times that reported by the Survey. When insurance data on reserves for future payments are also considered, the estimate of missed work days increases to 16,103,398, a figure 8.58-fold greater than that obtained for Michigan in the Survey. The Michigan data suggest that the national Survey may have failed to identify almost 373 million of 421 million missed work days in the private sector that have resulted, or will result, from 1986 occupational injuries. The present federal/state system for estimating occupational injury severity by measuring lost work days seriously underestimates the magnitude of the problem. The current policy of obtaining incidence and severity data from the same Survey should be reconsidered. We recommend that national estimates of injury severity be obtained from representative states by using state compensation data and that such estimates be used to evaluate current prevention and rehabilitation strategies. The redesigned occupational safety and health Survey (ROSH Survey) should be revised to permit linkage to compensation data. PMID- 8427254 TI - Fatal occupational accidents in Ontario, 1986-1989. AB - We examined 470 fatal occupational accidents in Ontario, 1986-1989, that met eligibility criteria. Homicides and most accidents on public roads were excluded. Information was obtained from coroners' files and records of the provincial Ministry of Labour. Levels of alcohol likely to produce impairment were found in six subjects (2% of the two-thirds of fatalities tested). Cannabis was detected in 3.9% of cases (17% of those tested), but other illegal drugs were not found. Recommendations of coroner's juries showed that organizational factors were considered relevant on many occasions, although language and literacy were rarely mentioned. The incidence rate rose steadily with age. Other data items were examined, although, because of missing information and/or lack of denominator data for many of them, the conclusions that can be drawn are limited. Among these tentative findings was that more fatal accidents occurred in the first half of the shift than in the second half. PMID- 8427255 TI - Job factors, radiation and cancer mortality at Oak Ridge National Laboratory: follow-up through 1984. AB - A previous study of mortality among white men hired at Oak Ridge National Laboratory between 1943 and 1972 (n = 8,318) revealed an association between low dose external penetrating ionizing radiation and cancer mortality in follow-up through 1984. The association was not observed in follow-up through 1977. This report considers the role of possible selection and confounding factors not previously studied. Control for hire during the World War II era and employment duration of less than 1 year had little effect on the radiation risk estimates. Risks associated with length of time spent in 15 job categories were considered as proxies for the effects of other occupational carcinogens. Adjustment for employment duration in each job category one at a time produced only small changes in the radiation risk estimate. Adjustment for potential exposures to beryllium, lead, and mercury also had little effect on the radiation risk estimates. These analyses suggest that selection factors and potential for chemical exposure do not account for the previously noted association of external radiation dose with cancer mortality. However, power to detect effects of chemical exposures is limited by a lack of individual exposure measures. PMID- 8427256 TI - A mortality study of cobalt production workers: an extension of the follow-up. AB - The follow-up of a cohort of workers employed in an electrochemical plant producing cobalt and sodium, previously studied from 1950-1980, has been extended from 1981-1988. The standardized mortality ratio (SMR) for all causes of death was 0.85 (95% confidence interval [CI] = 0.76-0.95, 309 observed) for the whole cohort, and 0.95 (95% CI = 0.83-1.08, 247 observed) for the subcohort of workers born in France. With regard to lung cancer mortality among cobalt production workers, which is the main objective of the study, the SMRs were, respectively, 0.85 (95% CI = 0.18-2.50, 3 observed) and 1.16 (95% CI = 0.24-3.40, 3 observed). Neither did any excess of mortality from diseases of the circulatory and of the respiratory systems appear among cobalt production workers. Maintenance workers, however, exhibited high SMRs for lung cancer, reaching statistical significance for duration of exposure and time since first exposure > or 30 years. This study does not support the hypothesis of a relationship between lung cancer and cobalt exposure. PMID- 8427257 TI - Occupational exposure to phenoxy herbicides and chlorophenols and cancer mortality in The Netherlands. AB - As part of the "IARC International Register of Persons Exposed to Phenoxy Herbicides and Contaminants," a cohort of workers who manufacture and prepare chlorophenoxy herbicides was recruited in The Netherlands. The cohort comprised 2,310 workers from two plants, operated by different companies, who were followed during the periods 1955-1985 and 1965-1986, respectively. In 1963, there had been an industrial accident in one factory with concomitant release of dioxin into the environment. Loss to follow-up was 3%. Mortality data on 963 exposed and 1,111 nonexposed men were evaluated by external and internal comparison. Compared with national rates, total mortality (94 deaths, standardized mortality ratio [SMR] = 101; 95% confidence interval [CI], 82-124) and cancer mortality (31 deaths, SMR = 107; 95% CI, 73-152) for exposed workers were not significantly increased. A statistically insignificant increase was observed for non-Hodgkin's lymphoma (2 deaths, SMR = 299; 95% CI, 36-1,078). No cases of soft-tissue sarcoma were encountered. There was no increase in either total mortality (25 deaths, SMR = 111; 95% CI, 72-163) or cancer mortality (10 deaths, SMR = 137; 95% CI, 66-252) among the 139 workers probably exposed to dioxins during the 2,4,5 trichlorophenol production accident or the subsequent clean-up operations. Compared with nonexposed workers, exposed workers did not exhibit a higher total mortality (rate ratio [RR] = 1.28; 95% CI, 0.89-1.82). Mortality due to all cancers (RR = 1.7; 95% CI, 0.9-3.4) and respiratory cancer (RR = 1.7; 95% CI, 0.5 6.3) was insignificantly elevated. These findings suggest that the increases in cancer mortality among workers exposed to phenoxy herbicides and chlorophenols may be attributable to chance. Lack of power prevented evaluation with respect to specific cancers. PMID- 8427258 TI - Evaluation of risks for non-Hodgkin's lymphoma by occupation and industry exposures from a case-control study. AB - The etiology of non-Hodgkin's lymphoma (NHL) is not well understood. To develop hypotheses on causes of this tumor, data from a population-based case-control interview study of 1,867 white men (622 cases and 1,245 controls) in Iowa and Minnesota conducted during 1980-1983 were examined. Subjects, or their next of kin, were interviewed to obtain information on agricultural exposures, work history, medical conditions, and family history. This analysis focuses on risks of NHL by occupation, by industry, and by selected exposures. Although many comparisons were made, few significant associations were observed. Small numbers and limitations in exposure assessment, however, would tend to reduce opportunities to detect associations. The strongest finding was with various occupations that work in metals and metal products. The analysis by exposure estimates also uncovered a significant association with metals, but risks did not increase with estimated intensity of exposure. Slightly elevated risks were also noted among persons employed as painters and construction workers, agricultural and forestry workers, printers and typesetters, funeral directors and embalmers, and dry cleaners. Although the overall risks for benzene and other solvents were small, they increased slightly with level of assigned exposure. Although some associations may be due to chance, several of these occupations and industries have been linked to lymphoma in other investigations and deserve further attention. PMID- 8427259 TI - Mortality and cancer incidence among PVC-processing workers in Sweden. AB - The mortality pattern and the cancer incidence were investigated among 717 men who had been employed for at least 3 months during 1964-1974 in three Swedish PVC processing plants. The mortality was followed 1964-1986 and the cancer incidence 1964-1984. Expected figures were calculated from Swedish national rates. Among Swedish citizens, the observed mortality and cancer incidence was close to the expected in most diagnoses. Among immigrants, mostly from Finland, there was a marked excess of circulatory deaths. This finding was probably due to the higher incidence of coronary mortality in Finland compared to Sweden. In the whole cohort, five cases of malignant melanoma had occurred as compared to 1.5 expected (SMR = 3.4, 95% confidence limit 1.1-7.9). This may be due to chance but merits further investigation since an increased incidence of malignant melanoma has previously been found among Norwegian PVC-manufacturing workers. PMID- 8427260 TI - Socioeconomic and health status of electronics workers employed in organized industry. AB - To study their socioeconomic and health status, 1,770 workers in the organized electronics industry in India were surveyed. Mean age of the workers employed in this industry was 32.5 +/- 6.01 years. The average per capita income/month was Rupees 333, indicating a higher socioeconomic status, compared with the status of the national population, as well as compared with their counterparts in the unorganized sectors. Respiratory symptoms and impairment rates were significantly higher in workers exposed to soldering fumes. The high prevalence of congested or inflamed throat was also related to chemical exposure specially among solderers and workers exposed to metal oxides. The ocular symptoms and signs were also related to chemical exposure. Musculoskeletal disorders were related to erratic ergonomic postures. PMID- 8427261 TI - Cotton dust and gram-negative bacterial endotoxin correlations in two cotton textile mills. AB - Exposure to cotton dust is known to cause both acute and chronic respiratory illness. A specific pattern of symptoms called byssinosis is well described to occur among workers in the cotton processing (e.g., yarn preparation) industry. Recent studies have implicated Gram-negative bacterial endotoxin as one of the agents responsible for acute, and possibly chronic, respiratory illness. Laboratory experiments using a model cardroom have found poor correlations between airborne dust and associated endotoxin. This study reports the results of vertical elutriated dust and endotoxin levels in 11 work areas of 2 cotton textile mills in 1986 in Shanghai, China. The overall correlation between dust and endotoxin was strong, rs = 0.66 and 0.79 (p < 0.0001) for mills 1 and 2, respectively. The dust-endotoxin correlation was relatively poor in early yarn preparation in the workshops and improved in the later preparation areas. Our findings suggest that in these mill settings, dust and endotoxin levels may be well correlated in most work areas. Therefore, dust may be a useful index for monitoring populations employed in the cotton textile industry throughout the world. Additional field studies need to be performed which consider the various determinants of dust and endotoxin levels. PMID- 8427262 TI - Childhood cancer and paternal exposure to ionizing radiation: preliminary findings from the Oxford Survey of Childhood Cancers. AB - Paternal occupational data already collected as part of the Oxford Survey of Childhood Cancers have been reviewed. Information on occupations during or before the relevant pregnancy was sought for 15,279 children dying from cancer in England, Wales, and Scotland in the period 1953-81, and for an equal number of matched controls. Estimates were made for paternal exposure to human-made external ionizing radiation in the six months before conception of the survey child--as judged from job histories and dates of birth. Assessments were also made for potential exposure to unsealed sources of radionuclides. Of the eight fathers placed in the highest dose group (> or = 10 mSv, external radiation), four were cases and four were controls. For the second dose group (5-9 mSv), the corresponding numbers were eight and four, and for the lowest exposed group (1-4 mSv), they were 55 and 42. There were 27 case fathers with potential exposure to radionuclides and only 10 control fathers. The independent effects of the two radiation variables were assessed by means of multiple logistic regression. Relative risks for estimated doses of external radiation were close to unity, but for radionuclide exposure the relative risk was 2.87 (95% CI = 1.15-7.13). These preliminary findings suggest that paternal exposure to radionuclides is a more likely risk factor for childhood cancer than exposure to external radiation. PMID- 8427263 TI - Schneeberg lung disease and uranium mining in the Saxon Ore Mountains (Erzgebirge). AB - The so-called Schneeberg lung disease is a form of bronchial or alveolar carcinoma caused by the effects of the radioactive gas radon and of its radioactive short half-life daughter products. This type of radiation-induced occupational cancer is the most common and the most important radiation injury among workers occupationally exposed to ionizing radiation. There have been many deaths from lung cancer, especially in the Soviet uranium mines in the Erzgebirge of Saxony in the former German Democratic Republic. The history of disease in these miners extends over five centuries; the first observations of their health hazard start in the Middle Ages. The discovery of the lung cancer component was made toward the end of the nineteenth century, and the suspicion that a connection might exist between this cancer type and exposure to ionizing radiation was voiced at the beginning of the twentieth century. In the first half of this century, further research was carried out on this disease in the Schneeberg area of the Erzgebirge. Before the end of World War II, guidelines were set up to define the acceptable limits of radon exposure in the ore mines of Saxony. After World War II, the American uranium mines in the Colorado Plateau used the German research results as a basis for working out their own radiation protection standard. The uranium mines under Soviet occupation in the former GDR, on the contrary, paid no attention to these research findings. For many years, no precautions were taken for the miners' working conditions. The consequence of this serious omission was an estimated 9,000 fatal cases of lung cancer among these underground miners. High concentrations of radon are to be found in indoor air of homes in some districts of the Erzgebirge, suggesting an increasing lung cancer risk for the local inhabitants. The significance of this finding is evaluated. PMID- 8427264 TI - Stability of ondansetron hydrochloride injection in extemporaneously prepared oral solutions. AB - The stability of ondansetron hydrochloride in extemporaneously prepared oral solutions containing orange juice, cola, or cherry syrup was determined. Solutions were prepared by adding ondansetron hydrochloride to orange juice, cola, or cherry syrup to produce ondansetron concentrations of 0.267 and 0.067 mg/mL in orange juice or cola and 0.533 mg/mL in cherry syrup. The ondansetron concentration in orange juice and cola solutions was assayed at the time of preparation and at 30 minutes and one hour. The cherry syrup solution was stored at both 3-5 and 25-27 degrees C, with the ondansetron concentration being determined at the time of preparation and daily for seven days. All the solutions were prepared in triplicate. Ondansetron concentrations were measured by stability-indicating high-performance liquid chromatography. At each time interval, the mean ondansetron concentration remained > or = 97% of the initial measurement for all solutions. The appearance and color of the solutions did not change. Ondansetron hydrochloride was stable for at least one hour in orange juice or cola and at least seven days in cherry syrup. PMID- 8427265 TI - Use of an antinausea drug by a pregnant woman. PMID- 8427266 TI - Managing pharmacy information systems. AB - The current state of pharmacy information systems is described, and a future direction for information management efforts is proposed. Today's pharmacy information systems are supported by excellent hardware and software. Systems are less expensive, more responsive, more flexible, and smaller than ever before. However, we have been ineffective at tailoring information systems to the pharmaceutical-care needs of patients. Instead, there is overemphasis on traditional requirements to support unit dose drug distribution, i.v. admixture preparation, and admission-discharge-transfer and billing systems. Deficiencies of current systems include a lack of real-time interfaces among hospital systems; redundancy of effort by users; ineffective capturing, reporting, and integration of data; and a cumbersome order-entry process. A vast amount of information is circulating; the challenge is to focus on those clinical and financial data on which patient-care decisions can be based. Each hospital pharmacy should develop a clinical information management strategy. Existing systems should be examined to determine which functions are not being used and which could be enhanced without major software modifications; enhancements might include routine entry of drug allergy information so that all orders may be screened and the use of networks and interfaces to provide medication profiles to physicians and nurses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427267 TI - Evaluating and selecting an information system, Part 1. AB - Initial steps in the process of evaluating and selecting a computerized information system for the pharmacy department are described. The first step in the selection process is to establish a steering committee and a project committee. The steering committee oversees the project, providing policy guidance, making major decisions, and allocating budgeted expenditures. The project committee conducts the departmental needs assessment, identifies system requirements, performs day-to-day functions, evaluates vendor proposals, trains personnel, and implements the system chosen. The second step is the assessment of needs in terms of personnel, workload, physical layout, and operating requirements. The needs assessment should be based on the department's mission statement and strategic plan. The third step is the development of a request for information (RFI) and a request for proposal (RFP). The RFI is a document designed for gathering preliminary information from a wide range of vendors; this general information is used in deciding whether to send the RFP to a given vendor. The RFP requests more detailed information and gives the purchaser's exact specifications for a system; the RFP also includes contractual information. To help ensure project success, many institutions turn to computer consultants for guidance. The initial steps in selecting a computerized pharmacy information system are establishing computerization committees, conducting a needs assessment, and writing an RFI and an RFP. A crucial early decision is whether to seek a consultant's expertise. PMID- 8427268 TI - Computerized services in hospital pharmacy departments. PMID- 8427269 TI - ASHP accreditation standard for pharmacy technician training programs. American Society of Hospital Pharmacists. PMID- 8427270 TI - Draft statement on pharmaceutical care. ASHP Council on Professional affairs. American Society of Hospital Pharmacists. PMID- 8427271 TI - FDA proposes more stringent pediatric-use labeling of prescription drug products. PMID- 8427272 TI - Congress compromises on Medicaid drug pricing; 'best price' excludes VA, others. PMID- 8427273 TI - Illinois supreme court says pharmacists' duty to warn is voluntary. PMID- 8427274 TI - HIV-infected persons say health-care workers discriminate. PMID- 8427275 TI - Market for antimicrobials expected to double by 1997. PMID- 8427276 TI - How to enhance intradepartmental communication. PMID- 8427277 TI - Obtaining continuing education when personal time is limited. PMID- 8427278 TI - Ensuring accuracy in the use of automatic compounders. PMID- 8427279 TI - Living positively with HIV. PMID- 8427280 TI - Filter clogging caused by albumin in i.v. nutrient solution. PMID- 8427281 TI - Filter clogging caused by albumin in i.v. nutrient solution. PMID- 8427282 TI - Patient advocate. PMID- 8427283 TI - Projecting future drug expenditures--1993. AB - The effects of inflation, generic competition, market introduction of new drug entities, and recent legislation on forecasting drug expenditures are discussed. Inflation as it relates to pharmaceutical prices has been decreasing over the past couple of years. The U.S. Bureau of Labor Statistics reported a 6.9% increase in the Producer Price Index for drugs and pharmaceuticals in 1991, diminishing to a 4.5% increase for part year 1992. Pharmaceutical industry analysts predict overall annual inflation rates for pharmaceuticals in 1993-94 will range from 2% to 11%. Explanations for the recent low inflation rates and the wide variance among analysts may include the uncertainty of future government regulation on price increases and the backlog of FDA approvals for biotechnologically derived agents. To evaluate generic competition, information on patent or market exclusivity expiration can be used. The price of a generic drug may be 60-70% of the brand price at market introduction, but it usually stabilizes at approximately 50% of the list price. Predicting the market entry of new drug products is difficult and requires monitoring of (1) filing dates for new drug applications (NDAs) and (2) changes within the FDA approval process. According to an FDA report, the mean time to approval for an NDA in 1991 was 28.5 months and for a new molecular entity was 30.03 months. These figures represent little change from the previous five years.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427284 TI - Comprehensive pharmaceutical services for pediatric patients. AB - The development of comprehensive pharmaceutical services for pediatric patients at a tertiary-care teaching hospital is described. A team of three staff pharmacists, a clinical specialist, and supportive personnel was formed. A pediatric pharmacy, operating from 0800 to 2100 daily, was created in a separate area of the central pharmacy to focus on potential problems with pediatric dosage calculations and drug administration. The team staffs the pediatric pharmacy for 75% of its day and evening shifts. The pharmacy prepares the 24-hour unit dose supply for each nursing unit, processes new orders, and provides drug information and problem-solving services. Clinical services are provided by the decentralized pharmacist, a rotating member of the team who makes rounds each day to the pediatric nursing units to review patient charts, provide medication information, and answer questions. The pediatric clinical specialist conducts educational programs, provides consultations, maintains reference materials, monitors pharmacokinetic evaluations, reviews medication communication forms, and assists in developing medication administration procedures. The creation of a pediatric pharmacy and a pediatric pharmacy team that coordinates both dispensing and clinical functions has made it possible to provide comprehensive pharmaceutical services to pediatric patients. PMID- 8427285 TI - Development of standards for providing prescription drugs by mail. AB - The development and implementation of practice standards for a mail-service pharmacy are described. Literature, licensing regulations, and existing national practice standards were reviewed. Selected mail-service pharmacies were surveyed on their practice standards. Patients who had received prescription drugs by mail from a university hospital pharmacy were asked their opinions of the service in a telephone survey. Based on the information gathered, a set of practice standards was drafted. The draft was considered by a panel of experts who reached a consensus on mail-service pharmacy practice standards. The standards were implemented at a university hospital's outpatient pharmacy. Standards of practice for mail-service pharmacies were developed and applied at a university hospital outpatient pharmacy. Other pharmacies may find the standards useful. PMID- 8427286 TI - Pharmacist management of a hyperlipidemia clinic. AB - A hyperlipidemia clinic in which a pharmacist provides primary care is described. The clinic was established at a Veterans Affairs medical center in January 1990. A pharmacist performs limited physical assessments, refers patients to other clinics as necessary, orders laboratory and diagnostic tests, and selects and monitors the use of lipid-lowering medications. Interventions are performed according to the pharmacist's clinical judgment; there is no set protocol. Recommendations for patient management are approved by an attending physician, who prescribes the antilipemic drugs. The pharmacist teaches patients about hyperlipidemia, the impact of diet and life-style, and the mechanism of action, administration, and adverse effects of the antilipemics prescribed. The pharmacist also monitors compliance, laboratory test values, and the response to treatment. Treatment is modeled after the recommendations of the National Cholesterol Education Program. If a patient has not achieved the targeted cholesterol concentration after receiving dietary therapy for three months, further education about diet and lifestyle is provided. If, after three more months, the cholesterol level remains high, drug therapy is begun. Four antilipemic drugs--colestipol, gemfibrozil, lovastatin, and niacin--are used in the clinic. Since it began operating, the clinic has enrolled 284 patients. Compliance with the lipid-lowering agents has ranged from 43% to 100%. Adverse effects have accounted for the majority of cases of noncompliance. A pharmacist provides primary care for patients with hyperlipidemia in an ambulatory-care clinic. PMID- 8427287 TI - Pharmacist assessment of drug allergies. AB - The impact of pharmacist-conducted drug allergy assessment on patient allergy labeling and subsequent drug therapy is reported. Pharmacists at an acute-care teaching hospital interviewed selected patients to verify their drug allergy status. If an interview resulted in the removal of an allergy label from the patient's allergy profile, the patient's medication profile was monitored daily to detect any subsequent use of the drug. If the targeted drug was prescribed, the patient's clinical status was assessed to determine the effect of the prescribed drug. A total of 606 drug allergy assessments were conducted in 358 patients. The allergy label was removed after 169 (28%) of the assessments; at least one label was removed for 121 (34%) of the patients. Fourteen patients (4%) received a targeted drug or drugs. Four of these patients had signs of clinical improvement. No adverse allergic reactions were observed in patients who received targeted agents. By interviewing patients, pharmacists identified and removed inappropriate drug allergy labels, enabling patients to receive medication that otherwise would have been withheld. PMID- 8427288 TI - Ketorolac injection use in a university hospital. AB - A drug-use evaluation (DUE) of ketorolac injection is described. Pharmacists at a 663-bed university hospital saved all new orders for ketorolac injection (except those from the operating, recovery, and emergency rooms) for five weeks. If an order was not in compliance with the DUE criteria, the DUE pharmacist contacted the prescriber to recommend a change. Fifty-two orders were evaluated. The daily dose exceeded that in the criteria in seven patients (13%). Conditions under which the product should be used with extra caution were present in eight patients (15%). Twelve patients (23%) received the drug for more than five days. Five patients (10%) had possible ketorolac-associated adverse effects. The pharmacist's concurrent interventions improved prescribing for correct dosage and facilitated discontinuation of the agent in high-risk patients. After follow-up educational efforts, a second audit in 28 patients showed further improvements in prescribing for correct dosage, but 25% of the patients received the drug for more than five days. No adverse effects were reported. A 72-hour automatic stop order policy was implemented, and the fraction of patients receiving the drug for more than five days fell to 3% (of 223 pharmacy profiles reviewed). Three patients had apparent ketorolac-induced adverse effects. Educational efforts and adoption of a 72-hour stop-order policy decreased the duration of ketorolac injection therapy and the use of inappropriately large dosages and may have decreased the frequency of ketorolac-induced adverse effects. PMID- 8427289 TI - Case report: high-dose intravenous immunoglobulin as therapy for thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura is a clinical syndrome defined by the pentad of thrombocytopenia, microangiopathic hemolytic anemia, fever, and renal and neurologic abnormalities. The pathogenesis of this syndrome remains enigmatic, though appropriate management usually involves plasma administration. The authors report on an alternative therapy, high-dose intravenous immunoglobulin, used in the patient after the failure of plasmapheresis. The implications and potential applications of this therapy are discussed. PMID- 8427290 TI - Case report: an unexpected intravenous pyelogram appearance in an azotemic patient. AB - The authors discuss the unusual intravenous pyelogram (IVP) findings in a patient with blunt abdominal trauma. Although the patient had apparent renal dysfunction, both nephrogram and pyelogram were seen clearly after a moderate dose of iothalamate. This may be explained by the fact that the underlying complete obstruction was relieved by his traumatic bladder rupture, prior to the contrast study. Alternatively, evidence of azotemia may be an artifact, resulting from resorption of urea and creatinine after rupture. Thus, the nephro-pyelogram may accurately reflect his renal function. PMID- 8427291 TI - Case report: malignant pericardial effusion as the initial manifestation of malignancy. AB - Although malignant pericardial effusion is a common complication of malignancy, it is rarely the initial manifestation. A case of a nonsmoking 36-year-old woman who had pericardial tamponade due to malignant pericardial effusion as the initial manifestation of malignancy is presented. Case reports of patients with malignant pericardial effusion as the initial manifestation of malignancy published since 1974 are reviewed. The clinical presentation, histology, and survival of these patients is compared with those cases published prior to 1974. Survival of patients with primary malignancy in the lung who present with malignant pericardial effusion has not improved. PMID- 8427292 TI - Case report: abnormal thyroid function tests in a patient and two normal volunteers treated with salsalate. AB - There are only three prior reports of abnormal thyroid function tests in patients who have received salsalate, the salicylate ester of salicylic acid. The authors report an elderly clinically euthyroid man who had thyroid function tests suggestive of central hypothyroidism while taking salsalate but whose thyroid tests returned to normal after the drug was discontinued. They also studied thyroid function tests, including free thyroxine (FT4) and reverse (T3), in two normal volunteers who took salsalate 750 mg twice daily for 1 week. In the normal subjects, total T4 and FT4 began to fall within 24 hours after the first dose of salsalate, and remained suppressed for at least 24 hours after the drug was discontinued. This rapidity of effect by salsalate is previously undescribed. There was also a fall in FT4, probably due to the use of diluted serum in the equilibrium dialysis FT4 assay. Because FT4 measurement using diluted serum or equilibrium dialysis may cause falsely low FT4 measurements, the authors believe ultrafiltration may be the only reliable method of measuring FT4 in these patients. PMID- 8427293 TI - Undifferentiated, overlapping, and mixed connective tissue diseases. AB - Undifferentiated connective tissue disease (UCTD) is a term used by many rheumatologists to define a group of diffuse connective tissue disorders that lack definitive characteristics of any particular well-defined disorder. Overlapping connective tissue disease is often used interchangeably with UCTD but they both refer to diseases that are in evolution before all the characteristic clinical and laboratory symptoms are manifested. However, the clinical features of some of the overlapping connective tissue diseases appear to be better defined. The classical one is mixed connective tissue disease, where features of systemic lupus erythematosus, progressive systemic sclerosis, and polymyositis may exist together with a positive anti-extractable nuclear antibody and high titers of anti-ribonuclear protein antibody. This review attempts to clarify the confusion between these terms. The problems in the clinical and laboratory diagnosis of common connective tissue diseases that coexist are addressed and treatment options discussed. The long-term implications of making a diagnosis of a definitive connective tissue disease before all the required criteria are met should be kept in mind because the patient may never develop the disease and yet be subjected to psychological, social, and economic hardships. PMID- 8427294 TI - Southwestern internal medicine conference: growth hormone--aging and osteoporosis. AB - Until recently, the use of growth hormone (GH) has been confined to the treatment of GH-deficient children. The advent of GH produced by recombinant DNA technology has increased the availability of GH. The increased availability of GH has made possible studies of the physiology and the possible therapeutic role of this hormone and its mediator insulin-like growth factor. One area where GH may play a therapeutic role is in the treatment of osteoporosis. This review will briefly summarize normal GH physiology and discuss age-related changes in GH and insulin like growth factor 1 (IGF-1) axis and how they may relate to age-related physiologic changes. Evidence for and against a possible therapeutic role for GH/IGF-1 in the treatment of age-related (senile) osteoporosis will be discussed. PMID- 8427295 TI - The role of the renin-angiotensin system in insulin sensitivity in normotensive subjects. AB - Nonobese patients with essential hypertension have been reported to be insulin resistant. Because blockade of the renin-angiotensin system has been demonstrated to improve insulin sensitivity in hypertensive patients, the authors evaluated the effects of angiotensin-II administration and of angiotensin-converting enzyme inhibition with enalapril on insulin-mediated glucose uptake in normotensive men. The results showed no change in insulin sensitivity with either 0.3 ng/kg/min or 1 ng/kg/min angiotensin-II administration or with enalapril therapy. The authors conclude that neither physiologic amounts of angiotensin-II nor converting enzyme inhibition alter insulin sensitivity in normotensive men. PMID- 8427296 TI - Will cytokines alter the treatment of myelodysplastic syndrome? AB - Myelodysplastic syndrome is a frustrating disorder, which until recently lacked effective treatment. Patients usually succumb to infection, bleeding complications, or progression to acute leukemia. Recombinant cytokines such as granulocyte macrophage-colony stimulating factor, granulocyte-colony stimulating factor, interleukin-3, and erythropoietin have been used to ameliorate the cytopenias associated with this disease. Small clinical trials in myelodysplastic syndrome patients, using cytokines with myeloid activity (G-CSF, GMCSF, IL-3), have shown consistent elevations in the white blood cell counts with little success in elevating hemoglobin or platelets. Erythropoietin is able to increase the hemoglobin in a small group of myelodysplastic syndrome patients. Future trials using combinations of these cytokines may lead to multilineage effects. PMID- 8427297 TI - Total lung capacity in asbestosis: a comparison of radiographic and body plethysmographic methods. AB - The effect of asbestosis on lung volumes was studied in 46 workers by measuring total lung capacity (TLC), forced vital capacity, vital capacity, and residual volume using two standard methods: body plethysmography and radiographic lung area. Nine men had neither irregular opacities of any International Labour Office profusion category nor pleural abnormalities; 9 had pleural abnormalities only, 19 had irregular opacities only; and 9 had pulmonary asbestosis with pleural plagues. Of the 28 with irregular opacities, 18 had moderate or advanced asbestosis (category 2/1 or greater). The radiographic method gave a mean TLC of 8.11 L and the body plethysmographic mean was 8.09 L. Thus, the radiographic and plethysmographic methods produced virtually identical mean values for TLC and for residual volume 3.84 L and 3.8 L and residual volume/TLC, 47.3 and 47.3, respectively. Lung volumes measured by body plethysmography and by x-ray area were the same in each of the four radiographic categories of asbestosis. The air trapping and normal or slightly elevated TLC, which characterize asbestosis in men who have smoked cigarettes, are revealed by both methods. The radiographic method, when adequate measures are taken to obtain full inspirations, effectively duplicates the body plethysmographic method but is quicker and simpler, particularly for field studies. PMID- 8427298 TI - Tubular site of the natriuresis after unilateral nephrectomy in the rat. AB - Unilateral nephrectomy (UNX) is followed by a prompt increase in sodium excretion from the remaining kidney. Recently, an important role for atrial natriuretic peptide (ANP) in mediating the UNX-associated natriuresis has been suggested. The present studies were undertaken to gain insight into the intrarenal mechanisms participating in the post-UNX natriuresis in circumstances in which the release or the action of endogenous ANP were suppressed by prior removal of the right atrial appendage and by administration of monoclonal anti-ANP antibodies, respectively. In anesthetized euvolemic untreated rats, UNX resulted in a twofold increase in urinary excretion of sodium (from 0.93 +/- 0.23 to 2.14 +/- 0.34 microE/min; p < 0.03), whereas glomerular filtration rate did not change significantly. Fractional excretion of lithium, an index of proximal tubular handling of sodium, increased from 30.7 +/- 3.4% to 39.4 +/- 4.0%, and fractional distal reabsorption of sodium decreased from 98.6 +/- 0.2% to 96.5% +/- 0.4% (both p < 0.006). Neither sham atrial appendectomy nor the administration of nonspecific antibodies affect the natriuretic response of the remaining kidney. The natriuretic response to UNX was abolished in right atrial appendectomized rats, as well as in rats receiving anti-ANP antibodies. Post-UNX changes in both proximal and distal tubular reabsorption of sodium were also suppressed in these animals. These observations indicate that ANP may be an important mediator of the natriuretic response to UNX and that the proximal and the distal part of the nephron contribute to the postnephrectomy natriuresis. PMID- 8427299 TI - Hyposomatomedinemia in quadriplegic men. AB - Many studies have shown that vigorous exercise acutely stimulates growth hormone (GH) release but the relative contribution of daily physical activity to maintaining the GH/somatomedin C (SmC) axis is not known. It has been reported that basal and post-exercise plasma SmC values are higher in physically conditioned young men than in sedentary men of similar age. To assess the effect of severe inactivity on the plasma SmC level, basal concentrations of this hormone were measured in patients with quadriplegia (QP) resulting from spinal cord injury (SCI). Venous blood samples were obtained after overnight fast in 41 QP men, ages 24-66, and compared with 119 healthy men of similar ages. Nonparametric analysis of variance showed SmC to be significantly lower in QP than in healthy men (p < .007). Plasma SmC below 0.35 U/ml in adults usually indicates little or no GH secretion by the pituitary gland. In QP, 46% of plasma SmC values were < 0.35 U/ml compared to 24% in the healthy group (p < .02). In both groups, an inverse relationship of SmC and increasing age was observed (p < .01). The data suggest that severe inactivity or SCI tend to cause hyposomatomedinemia. The latter endocrine alteration may contribute to the decrease in lean body mass and muscle atrophy of QP patients, and add further functional impairment to the original neurologic deficit. In addition, hyposomatomedinemia could increase the tendency for pressure sore formation and osteoporosis in SCI patients. PMID- 8427300 TI - Factors affecting expressed emotion in parents of ill and normal children. AB - In order to examine their relationship to Expressed Emotion, family environment and marital relationships were measured in the parents of disruptive, obsessive compulsive, and normal children. Satisfactory family and martial environment were found to be related to low Expressed Emotion, while achievement orientation in mothers and greater family conflict were found to be significantly related to high Expressed Emotion. PMID- 8427301 TI - Suicidality in hospitalized adolescents: relationship to prior abuse. AB - The effects of admission status, prior abuse, and the frequency and duration of both physical and sexual abuse on measures of suicidality for a sample of 117 hospitalized adolescents were investigated. Having been abused was found to have a significant association with the number of previous suicide attempts and to interact with the variable of admission status in measures of suicide ideation. PMID- 8427302 TI - Planned conception and infant functioning at age three months: a cross-cultural study. AB - The conceptions of 53 American and 54 Greek infants were categorized as planned or unplanned on the basis of mothers' reports. At age three months in both cultural groups, infants in the planned category showed higher levels of cognitive processing and attachment to their mothers than did infants in the unplanned category, as shown by their differential vocal responses to mothers versus a female stranger. PMID- 8427303 TI - The role of paternal variables in divorced and married families: predictability of adolescent adjustment. AB - The predictability of adolescent adjustment, based on teacher reports, in divorced and married families was examined in relationship to four variables: paternal mood, spouse-ex-spouse relationship, father-adolescent relationship, and parental visitation. Analyses indicated that the father-adolescent relationship was predictive of adolescent functioning in divorced families, and the spousal relationship and paternal mood were similarly predictive in married families. PMID- 8427304 TI - Childhood family environment and sexual abuse as predictors of anxiety and depression in adult women. AB - A study of 46 women with histories of childhood sexual abuse and a control group of 93 women without such histories showed an association between childhood sexual abuse and the women's symptoms of anxiety and depression, as well as their perceptions of their families of origin. Results also suggested that family conflict, control, and cohesiveness moderated the relationship between the childhood abuse and current symptoms of depression. PMID- 8427305 TI - The psychosocial climate of families with suicidal pre-adolescent children. AB - A structured interview measure of suicidal behavior and a questionnaire measure of family psychosocial climate were administered to 43 pre-adolescent psychiatric inpatients and their parents. Results showed that suicidal behavior tended to be associated with greater family conflict and with less family organization, cohesion, and achievement orientation. PMID- 8427306 TI - Mothers of children with spina bifida: factors related to maternal psychosocial functioning. AB - Maternal self-perceptions of well-being and of satisfaction and competence in the parental role were examined in 50 mothers of children diagnosed with spina bifida. Adult companionship and social support were found to be highly correlated with maternal satisfaction, while quality of family environment significantly predicted maternal competence and well-being. PMID- 8427307 TI - Young children of schizophrenic mothers: difficulties of intervention. AB - Casework by Danish local social agencies on behalf of 11 children of chronically ill schizophrenic mothers is retrospectively analyzed, along with documentation from psychiatric hospitals, consulting child specialists, and other health professionals. Findings point to a need for earlier and more precise assessment of the mother's parenting abilities as measured against the severity of her illness and the vulnerability of her child. PMID- 8427308 TI - Short-term family-based residential treatment: an alternative to psychiatric hospitalization for children. AB - A short-term treatment program designed as an alternative to psychiatric hospitalization for children and adolescents is described. The program utilizes a multidisciplinary professional team and specially trained individuals (mentors) who work with the child and the child's biological family in the context of a mentor's home. Admission, discharge, and follow-up data on a group of patients are reported. PMID- 8427309 TI - Clinical dilemmas: therapists treating therapists. AB - Psychotherapists who treat therapist-patients confront a range of intense, often unexpected, countertransferential feelings that threaten to intrude on the course of treatment. The experience of the treating therapist is examined and discussed with particular attention to implications for clinical practice. PMID- 8427310 TI - Research: the latest magic wand. PMID- 8427311 TI - Perceptions of professional competence: cross-disciplinary ratings of psychologists, social workers, and psychiatrists. AB - Psychiatrists, psychologists, social workers, and nurses rated members of the first three disciplines on effectiveness of intervention for six hypothetical clients representing a range of mental health problems. Results revealed within group bias, with perceptions that professionals sharing raters' own professional affiliation would be more helpful, expert, and warm, as well as the preferred recipient of referrals. Clinical social workers were rated highest on warmth by all raters but lowest on referral intent by raters from other disciplines. PMID- 8427312 TI - The vulnerability-stress model of schizophrenia: advances in psychosocial treatment. AB - Vulnerability models of schizophrenia are reviewed, along with psychosocial rehabilitation methods addressing functional abilities and social competence. Their relationship is discussed with a view to developing a framework in which biological and psychosocial approaches to schizophrenia can be integrated for purposes of effective clinical intervention. Such intervention is designed to improve social competence, cognitive appraisal, and coping skills for mediation of stress in vulnerable individuals. PMID- 8427313 TI - Childhood play in normality, pathology, and therapy. AB - Links are sought between concepts of play pertaining to normal development, pathology, and therapeutic interventions. To this end, the components and functions of play are delineated, the evolution of the concept of play is reviewed, an integrated course for its development is advanced, and a framework and guidelines for its assessment in the clinical context are offered. Issues concerning the role of play in therapy are discussed. PMID- 8427314 TI - Substance abuse with psychiatric illness in children and adolescents: definitions and terminology. AB - Substance abuse with psychiatric illness is commonly referred to as comorbidity, a term that lacks the specificity needed to further precision in research and clinical treatment, both of which are particularly important when dealing with children and adolescents. A review of the literature on this dual disorder in young people is followed by a discussion of diagnostic and treatment considerations and their implications for researchers and clinical practitioners. PMID- 8427315 TI - Sheltered care residence: ten-year personal outcomes. AB - A ten-year follow-up study of 393 seriously mentally ill sheltered-care residents examined the type and degree of handicap characterizing the sample and estimated effects of ten-year residence. Results showed higher levels of helper-supported social functioning and of physical and mental health, accompanied by significant reductions in independent social functioning. PMID- 8427316 TI - Mothers of sexually abused children: trauma and repair in longitudinal perspective. AB - Mothers whose children had been sexually abused reported experiencing serious psychological symptoms following disclosure of the abuse. Over a one-year period, their emotional status improved. Strong relationships between mothers' reports of their own and their children's symptoms were accompanied by persistent discrepancies between maternal and direct assessments of the children's emotional states. Findings suggest that addressing maternal distress is important to the study and treatment of child sexual abuse. PMID- 8427317 TI - Suicide: risk factors and the public health. PMID- 8427318 TI - The dissemination of smoking cessation methods for pregnant women: achieving the year 2000 objectives. AB - The smoking prevalence rate among adult women and pregnant women has decreased only 0.3 to 0.5% per year since 1969. Without a nationwide dissemination of efficacious smoking cessation methods based on these trends, by the year 2000 the smoking prevalence among pregnant women will be approximately 18%. This estimate is well above the US Department of Health and Human Services Year 2000 Objective of 10%. The US dissemination of tested smoking cessation methods could help an additional 12,900 to 155,000 pregnant smokers annually and 600,000 to 1,481,000 cumulatively to quit smoking during the 1990s. Dissemination could help achieve 31 to 78% of the Year 2000 Objectives for pregnancy smoking prevalence. (With dissemination, at best a 15% smoking prevalence during pregnancy, rather than the 10% objective, is likely to be observed.) Our results confirm a well-documented need for a national campaign to disseminate smoking cessation methods. PMID- 8427319 TI - Aggression, substance use, and suicidal behaviors in high school students. AB - OBJECTIVES: We sought to analyze the frequency and correlates of suicidal behaviors in a community sample of adolescents. METHODS: Information concerning suicidal thoughts and acts, aggressive behaviors, substance use and physical recklessness were collected with the 70-item self-report Youth Risk Behavior Survey from a statewide sample of 3764 South Carolina public high school students. RESULTS: Seventy-five percent of students reported no suicidal behaviors, 11% reported serious suicidal thoughts, 6.4% reported specific suicidal plans, 5.9% reported attempts not requiring medical care, and 1.6% reported attempts requiring medical care. All types of suicidal behaviors occurred more frequently in females than males. Odds ratios for aggressive behaviors and cigarette use were elevated across all categories of suicide behaviors, increasing in magnitude with severity of reported suicidal behavior. Substance use was associated with some but not all categories of suicidal behaviors. The relationships were most pronounced with the use of potentially more dangerous drugs. CONCLUSIONS: The results suggest that suicidal behaviors are not infrequent occurrences among adolescents and that they often coexist with other high-risk behaviors. Interventions designed to reduce suicidal behaviors should simultaneously address coexisting high-risk behaviors. PMID- 8427320 TI - Risk factors for early adolescent drug use in four ethnic and racial groups. AB - OBJECTIVES: It is widely believed that risk factors identified in previous epidemiologic studies accurately predict adolescent drug use. Comparative studies are needed to determine how risk factors vary in prevalence, distribution, sensitivity, and pattern across the major US ethnic/racial groups. METHODS: Baseline questionnaire data from a 3-year epidemiologic study of early adolescent development and drug use were used to conduct bivariate and multivariate risk factor analyses. Respondents (n = 6760) were sixth- and seventh-grade Cuban, other Hispanic, Black, and White non-Hispanic boys in the 48 middle schools of the greater Miami (Dade County) area. RESULTS: Findings indicate 5% lifetime illicit drug use, 4% lifetime inhalant use, 37% lifetime alcohol use, and 21% lifetime tobacco use, with important intergroup differences. Monotonic relationships were found between 10 risk factors and alcohol and illicit drug use. Individual risk factors were distributed disproportionately, and sensitivity and patterning of risk factors varied widely by ethnic/racial subsample. CONCLUSIONS: While the cumulative prevalence of risk factors bears a monotonic relationship to drug use, ethnic/racial differences in risk factor profiles, especially for Blacks, suggest differential predictive value based on cultural differences. PMID- 8427321 TI - The effects of intrauterine cocaine exposure in newborns. AB - OBJECTIVES: We sought to determine the effects of intrauterine cocaine exposure in newborns, in an inner-city population in which cocaine use during pregnancy was common. METHODS: During a 1-year period, 12.8% (361 of 2810) of all live singleton infants at Harlem Hospital in New York were identified as cocaine exposed, either by universal urine toxicologic screening or by maternal history. Cocaine-exposed infants were compared with a control group of 387 infants not known to be exposed to cocaine or other illicit drugs. RESULTS: Low birthweight (< 2500 g) was more common among cocaine-exposed infants (31% vs 10%), as was preterm birth (< 37 completed weeks of gestation) (32% vs 14%). In multivariate analyses controlled for demographic and life-style factors and duration of gestation, cocaine was associated with decreased birthweight (154 g), length (1.02 cm), head circumference (0.69 cm), and duration of gestation (0.74 weeks). The birthweight deficits were larger for infants born to mothers who used cocaine in combination with other drugs (195 g) and for infants born to mothers who specifically admitted using crack (200 g). CONCLUSIONS: Intrauterine cocaine exposure is linked with fetal growth retardation and shortened gestation in this population. PMID- 8427322 TI - Drug abuse and illicit drug use in Puerto Rico. AB - OBJECTIVES: Based on an epidemiologic field survey of community households in Puerto Rico, this study estimates the frequency of illicit drug use and clinically defined drug abuse and/or dependence syndromes. Results are compared with those from surveys on the United States mainland. Suspected risk factors are studied as well, with a special focus on childhood misbehavior. METHODS: Trained lay interviewers administered a Spanish Diagnostic Interview Schedule to 912 respondents aged 17 to 68 years who were selected by multistage probability sampling of island households. RESULTS: An estimated 8.2% of the population had a history of illicit drug use and 1.2% qualified for a standardized lifetime diagnosis of drug abuse, dependence, or both. An estimated 18.4% of the male drug users and 7.7% of the female drug users met criteria for drug abuse and/or dependence. A history of drug use was related to the diagnoses of alcohol abuse and/or dependence and antisocial personality, but few persons who had used illicit drugs at least once in their lifetime reported a history of receiving treatment for alcohol, drug, or mental health problems. CONCLUSIONS: The data were consistent with a suspected association between level of childhood misbehavior and occurrence of illicit drug use, even after statistical control for potentially confounding variables. PMID- 8427323 TI - Health education for pregnant smokers: its behavioral impact and cost benefit. AB - OBJECTIVES: A randomized trial (the Birmingham Trial II) was conducted to evaluate the behavioral impact of health education methods among 814 female smokers at four public health maternity clinics. METHODS: Four hundred patients were randomly assigned to an Experimental (E) Group, and 414 were assigned to a Control (C) Group. Self-reports and saliva cotinine tests confirmed smoking status at the first visit, at midpregnancy, and at end of pregnancy. RESULTS: The E Group exhibited a 14.3% quit rate and the C Group an 8.5% quit rate. A Historical Comparison (C) Group exhibited a 3.0% quit rate. Black E and C Group patients had higher quit rates than White E and C Group patients. A cost-benefit analysis found cost-to-benefit ratios of $1:$6.72 (low estimate) and $1:$17.18 (high estimate) and an estimated savings of $247,296 (low estimate) and $699,240 (high estimate). CONCLUSION: Health education methods are efficacious and cost beneficial for pregnant smokers in public health maternity clinics. PMID- 8427324 TI - Paternal smoking and birthweight in Shanghai. AB - OBJECTIVES: Although maternal active smoking has been established to be associated with fetal growth retardation, evidence of an effect of environmental tobacco smoke exposure on birthweight is still limited and inconclusive. This study addressed the relationship between prenatal environmental tobacco smoke exposure and birthweight and fetal growth retardation in Shanghai, China. METHODS: Data on 1785 full-term live-born normal infants of nonsmoking mothers were used from the Shanghai Birth Defects and Perinatal Death Monitoring conducted between October 1986 and September 1987. Environmental tobacco smoke exposure was defined as exposure to paternal smoking. RESULTS: Infants with environmental tobacco smoking exposure were, on average, 30 g lower in birthweight than nonexposed infants, after adjustment for gestational age, parity, maternal age, and occupation. CONCLUSION: Consistent with previous research, this study suggests that environmental tobacco smoking exposure may have a modestly adverse effect on birthweight. PMID- 8427325 TI - Cigarette smoking and the risk of diabetes in women. AB - OBJECTIVES: Noninsulin-dependent diabetes mellitus, a major risk factor for cardiovascular disease, is prevalent in more than 12 million Americans. A voluminous amount of data demonstrates that cigarette smoking is an important cause of cancer and coronary heart disease. However, the association between cigarette smoking and the risk of diabetes is virtually unexplored, especially in women. METHODS: We examined the association between smoking and the incidence of noninsulin-dependent diabetes mellitus among 114,247 female nurses who were free of diabetes, cardiovascular disease, and cancer in 1976. We collected exposure information and disease status prospectively for 12 years from biennially self administered questionnaires. RESULTS: Current smokers had an increased risk of diabetes, and we observed a significant dose-response trend for higher risk among heavier smokers. During 1,277,589 person-years of follow-up, 2333 women were clinically diagnosed with diabetes. The relative risk of diabetes, adjusted for obesity and other risk factors, was 1.42 among women who smoked 25 or more cigarettes per day compared with nonsmokers. CONCLUSIONS: These data suggest that cigarette smoking may be an independent, modifiable risk factor for noninsulin dependent diabetes mellitus. PMID- 8427326 TI - Self-exempting beliefs about smoking and health: differences between smokers and ex-smokers. AB - OBJECTIVES: The purpose of the present study was to examine the role of self exempting or cognitive dissonance-reducing beliefs about smoking and health. Such beliefs may hold important implications for the content and targeting of health promotion campaigns. METHODS: A survey of smokers and ex-smokers was conducted in western Sydney, Australia. Six hypotheses were tested. RESULTS: The principal findings were (1) that 27.9% of smokers and 42.1% of ex-smokers agreed that smokers were more likely than non-smokers to get five smoking-related diseases; (2) that for 11 of 14 beliefs tested, more smokers than ex-smokers agreed to a statistically significant degree; (3) that the median number of such beliefs agreed to by smokers was five, compared with three for ex-smokers; (4) that for only 5 of 14 beliefs was agreement expressed by more precontemplative smokers than smokers contemplating or taking action to quit; (5) that more than one in four smokers, despite agreeing that smokers are more likely than non-smokers to get five diseases, nonetheless maintain a set of self-exempting beliefs. CONCLUSIONS: Fewer smokers than ex-smokers accept that smoking causes disease, and smokers also maintain more self-exempting beliefs. Becoming an ex-smoker appears to involve shedding such beliefs in addition to accepting information about the diseases caused by smoking. PMID- 8427327 TI - Smoking cessation factors among African Americans and whites. COMMIT Research Group. AB - OBJECTIVES: This study was undertaken to explore smoking patterns and attitudes that influence smoking cessation and relapse among African Americans. METHODS: Baseline data from eight Community Intervention Trial for Smoking Cessation (COMMIT) sites were analyzed. RESULTS: Compared with Whites, African Americans who smoke less than 25 cigarettes per day were 1.6 times more likely to smoke within 10 minutes of awakening (a behavioral indicator of nicotine dependence), adjusting for education, age, and gender (OR = 1.2 for heavier smokers). African Americans reported a stronger desire to quit smoking and reported serious quit attempts in the past year. African Americans favored tobacco restrictions (they were 1.8 times more likely than Whites to view smoking as a serious community problem, 1.7 times more likely to favor restrictions on cigarette vending machines, and 2.1 times more likely to prohibit smoking in their car). African Americans were lighter/moderate, menthol smokers. CONCLUSIONS: African Americans find smoking socially unacceptable and are strongly motivated to quit, but their "wake-up" smoking may indicate high nicotine dependence, making abstinence difficult even for lighter smokers. PMID- 8427328 TI - Retail stores' compliance with a city no-smoking law. AB - OBJECTIVES: Laws restricting smoking in public places and workplaces can protect the public only if they are obeyed. We sought to assess compliance with a Cambridge, Mass, no-smoking ordinance. METHODS: We prospectively observed 174 retail stores 1 month before and 3, 11, and 24 months after the law took effect. At 24 months, we interviewed one employee per store. RESULTS: Full compliance with the law was low; at 24 months, only 4% of stores displayed the mandated sign and were free of smokers and smoke. Fewer than half the stores posted any no smoking sign. Sign prevalence increased over 2 years (22% to 41%, P < .001), but the frequency of smoke or smokers (13% and 10%, respectively, at baseline) did not change. According to employees interviewed at 24 months, 38% of stores illegally permitted customers or employees to smoke. These stores had more smoke and fewer signs than did stores prohibiting smoking. Compliance was poor in liquor and convenience stores. Employees who had been told how to handle customers' smoking were more likely to enforce the law. CONCLUSIONS: Compliance with a no-smoking law is not guaranteed. For retail stores, compliance may improve if stores adopt no-smoking policies, post signs, and teach employees to enforce the law. PMID- 8427329 TI - The implications of an epidemiological mistake: a community's response to a perceived excess cancer risk. AB - OBJECTIVES: The response of community residents to a perceived cancer excess may include changes in attitude, health-related behavior, and property values. In 1986, a cancer agency conducted a study of cancer incidence (1979 to 1983) in two suburbs of Edmonton, Alberta, and reported elevations on the order of 25% over expected for most sites. Reanalysis of these data several months later revealed an error. Correction brought the rates into line with Alberta as a whole and with other communities surrounding Edmonton. METHODS: We used public opinion trends and property value trends (during the period of concern) to study the two communities affected by the allegation of increased cancer risk. RESULTS: A survey of residents found significant differences at the time in health-related behavior and beliefs suggesting increased perception of personal, family, and community risk and modest changes in behavior. Real estate values in one community temporarily lost an average of $4000, or about 5% of total value, compared with a similar, adjacent housing market. CONCLUSIONS: The perception of an elevated cancer risk, in the absence of a true risk, may have a substantial negative effect on the affected community, both psychologically and economically. PMID- 8427330 TI - Symptoms of depression among blacks and whites. AB - OBJECTIVES: We sought to examine relationships between sociodemographic indicators of risk and depression symptoms within the Black and White populations. METHODS: In a national probability sample, differences in sex, age, marital status, religion, social class, employment status, urbanicity, and region were evaluated against a Center for Epidemiological Studies Depression Scale score of 16 or greater. Risk factors were identified within the two populations and compared between them. RESULTS: For both races, females were at greater risk for depression than males, and respondents who were formerly married or separated were at greater risk than those who were currently married. Major differences in patterns of risk were also found between the races. Blacks who were 30 to 39 years of age, belonged to non-Western religious groups, and lived in the West were at greater risk than comparable Whites. Blacks who were widowed, members of the middle and lower-middle class, and unemployed were at less risk. CONCLUSIONS: Similarities and differences in patterns of risk were evident. The nature of the differences suggests important divergence in sociocultural and economic experience. PMID- 8427331 TI - Socioeconomic status and prediction of ventricular fibrillation survival. AB - OBJECTIVES: The association between socioeconomic status and cardiac arrest is less well known than some other associations with cardiac arrest. We used property tax assessments as a measure of socioeconomic status in a study of victims of out-of-hospital cardiac arrest found in ventricular fibrillation. METHODS: We studied patients attended by the Seattle Fire Department's emergency medical services system between May 1986 and August 1988. During the period studied, 356 episodes met the study criteria; 114 (32%) of these patients survived without major neurologic deficit. Residential property tax assessments were available for 253 of the patients. RESULTS: After adjustments were made for age, witnessed collapse, bystander-initiated cardiopulmonary resuscitation, time from call to paramedic arrival, activity, location of collapse, and chronic morbidity, an association of survival with greater assessed value per living unit was observed. An increase of $50,000 in value per unit was associated with a 1.6 fold increase in survival rate. CONCLUSIONS: Not only are persons in the lower socioeconomic strata at greater risk for cardiac mortality, but they are also less likely to survive an episode of out-of-hospital cardiac arrest. PMID- 8427332 TI - Smoking and suicide among nurses. AB - Current evidence suggests a strong positive correlation between cigarette consumption and depression; this study examined the relationship between cigarettes and suicide. Over 100,000 predominantly White, middle-aged, female registered nurses were followed via biannual questionnaires from 1976 through 1988. Respondents smoking 1 through 24 cigarettes per day had twice the risk and those smoking 25 or more cigarettes four times the risk of committing suicide, compared with those who had never smoked. Although no information on causation was available, this paper links cigarettes to another major health problem. PMID- 8427333 TI - A stop-smoking telephone help line that nobody called. AB - The purpose of this study was to evaluate the reach of a smoker's hotline that provided a variety of services to over 2100 health maintenance organization members. Formative evaluation procedures identified topics of concern, and repeated promotions advertised the service via multiple channels. Excluding a special giveaway promotion, an average of less than three calls per month were made during the 33 months of operation. To be cost-effective, smoker's hotlines should be offered to large populations and should be consistently and intensively publicized. PMID- 8427334 TI - Workplace smoking policies in New Jersey businesses. AB - A New Jersey law enacted in 1985 requires smoking policies for work sites with 50 or more employees. In 1988, we surveyed a sample of New Jersey businesses to determine their smoking policies. Eighty-three percent of respondents with 50 or more employees reported implementing restrictive smoking policies. Of respondents with 2 to 49 employees, 35% had implemented policies. Our findings identify some of the factors other than a legal mandate that lead employers to restrict smoking in the workplace. PMID- 8427335 TI - Patterns of drug use among Cuban-American, African-American, and white non Hispanic boys. AB - This study examined initiation into drug use during grade school years in a sample of Cuban-American, Black, and White non-Hispanic students in the greater Miami, Fla, area. Findings indicate that first use of alcohol occurs in fifth grade and cigarettes in sixth grade for all subgroups except White non-Hispanics, who peak in the fifth grade. White non-Hispanics had the highest life-time levels of alcohol and cigarette use. Foreign-born Cuban Americans had a lower lifetime prevalence of alcohol and cigarette use than US-born Cuban Americans. Higher acculturation level was related to first use of alcohol. One important implication of this study is that alcohol interventions should begin no later than third grade and smoking interventions no later than fourth grade. PMID- 8427336 TI - Male transvestite prostitutes and HIV risk. AB - Human immunodeficiency virus (HIV)-1, syphilis, and hepatitis B prevalence and associated risk factors were assessed among male transvestite prostitutes. Structured street-level interviews were conducted with 53 respondents in Atlanta, Ga, from July 1990 through July 1991. Test results from serum samples revealed that 68% were seropositive for HIV-1, 81% had seromarkers for syphilis, and 80% had seromarkers for hepatitis B. Univariate logistic regression analysis indicated that seromarkers for syphilis and Black race were the primary factors associated with HIV-1 infection. The results show that transvestite prostitutes are a heterogenous population and distinct from nontransvestite prostitutes; specific outreach is thus needed. Targeted interventions should address the sexual and drug-use-related HIV risk behaviors of transvestite prostitutes. PMID- 8427337 TI - The physical activity of fifth-grade students during physical education classes. AB - One hundred fifty-seven fifth-grade students in 20 of the 355 elementary schools in one Texas county were systematically observed during physical education classes. On average, the students spent 8.5% of class time in moderate to vigorous physical activity, 23.3% in minimal activity, and 68.1% in sedentary activity. None of the schools averaged 20% of class time in moderate to vigorous physical activity. The levels of physical activity observed are substantially lower than the levels called for in national health objectives. PMID- 8427338 TI - Left-handedness and mortality. AB - We examined mortality associated with handedness in two ways. A simulation using national data suggests that lower mean age at death among left-handed persons, previously offered as evidence of higher mortality, can be explained exclusively by the age distribution of laterality. Second, empiric evidence from a 6-year cohort study of 3774 older adults from East Boston, Massachusetts, demonstrates that left-handedness is not associated with mortality (relative odds = 1.04, 95% confidence interval = 0.79, 1.36). PMID- 8427339 TI - The effect of fish oil supplements on blood pressure. AB - We conducted a double-blind, placebo-controlled crossover study to determine the effects of fish oil supplementation on blood pressure in middle-aged men. Subjects were randomly assigned to consume either 20 g of fish oil or safflower oil for 12 weeks and then consume the other oil for an additional 12 weeks after a 4-week washout period. We found no significant changes from the pretreatment value in systolic or diastolic blood pressure with the use of fish oil supplements. In addition, there were no significant differences in the posttreatment blood pressures comparing the fish and safflower oil phases of the study. PMID- 8427340 TI - The nitrate contamination of private well water in Iowa. AB - The State-Wide Rural Well-Water Survey was conducted between April 1988 and June 1989. About 18% of Iowa's private, rural drinking-water wells contain nitrate above the recommended health advisory level (levels of NO3-N greater than 10 mg/L); 37% of the wells have levels greater than 3 mg/L, typically considered indicative of anthropogenic pollution. Thirty-five percent of wells less than 15 m deep exceed the health advisory level, and the mean concentration of nitrate nitrogen for these wells exceeds 10 mg/L. Depth of well is the best predictor of well-water contamination. Individually, NO3-N levels of more than 10 mg/L occurred alone in about 4% of the private wells statewide; pesticides were present alone in about 5%. Total coliform positives occurred alone at 27% of the sites. In a cumulative sense, these three contaminants were detected in nearly 55% of rural private water supplies. PMID- 8427341 TI - Benjamin Rush's educational campaign against hard drinking. AB - More than 200 years ago, during a period of unprecedented production and consumption of distilled alcoholic beverages in the United States, Benjamin Rush launched a health education campaign that warned the public about the hazards of such beverages. He corrected erroneous notions about their presumed beneficial effects and accurately described more than a dozen alcohol-related health problems. Although the temperance movement has had a tumultuous history in the United States, the origin and long-standing tradition of temperance as a health promotion activity needs to be recognized. PMID- 8427342 TI - Children and firearms: a gunshot injury prevention program in New Mexico. PMID- 8427343 TI - Harnessing the energy of the mass media: HIV awareness in Dallas. PMID- 8427344 TI - A pilot study of HIV-infected immigrants. PMID- 8427345 TI - The HIV rates of injection drug users in less-populated areas. PMID- 8427346 TI - Maternal age and cesarean delivery rate in Shanghai. PMID- 8427347 TI - Measuring the effects of vitamin A supplementation. PMID- 8427348 TI - Multidisciplinary findings on socioeconomic status and health. PMID- 8427349 TI - Computerized multiple cause-of-death information available from NCHS. PMID- 8427350 TI - The predictive value of radiographs in the evaluation of unilateral and bilateral anterior cruciate ligament injuries. AB - A review of 250 cases of surgical reconstruction of the anterior cruciate ligament identified 24 patients with bilateral complete tears of the anterior cruciate ligament. Twenty of these patients had previous reconstruction of one anterior cruciate ligament before rupture of the opposite ligament. Twelve injuries occurred during the same activity that was responsible for the initial opposite injury. The average time between surgical reconstruction and rupture of the opposite ligament was 29.3 months (range, 3 to 103). No significant demographic differences existed between patients with unilateral or bilateral ruptures of the anterior cruciate ligament. Standardized measurements of intercondylar notch height and width and medial and lateral femoral condyle height and width were performed on routine notchview height and width were performed on routine notchview radiographs of 31 knees of patients with bilateral injuries, 30 with unilateral injury, and 30 with no anterior cruciate ligament injury. Statistical analysis revealed no significant differences between the three groups when comparing absolute measures or any of eight mathematical ratios calculated from these measurements. We concluded that measurements of the intercondylar notch made from radiographs may not be reliable predictors of injury to the anterior cruciate ligament. We found no significant clinical or demographic differences between patients with unilateral or bilateral complete ruptures of the anterior cruciate ligament. PMID- 8427351 TI - Repair of distal biceps tendon ruptures in athletes. AB - Ten athletes with distal biceps tendon ruptures that had been anatomically repaired with a double-incision techniques were reviewed to determine their functional recovery. All of the patients were men, with an average age of 40 years (range, 25 to 49). Eight of the 10 patients were weight lifters or body builders, and 7 had participated on a competitive level at some point in their athletic careers. Six injured their dominant extremity, and 4 their nondominant extremity. Isokinetic muscle testing of supination and flexion was performed in 8 patients and the results were compared to a control group. Followup averaged 50 months (range, 12 to 105). Patients uniformly graded their subjective results as excellent, with a group mean rating of 9.75 on a 10-point scale. All athletes returned to full, unlimited activity. The contour of the biceps muscle was restored in all cases. Isokinetic muscle testing demonstrated that in those patients with a repaired dominant extremity, supination strength and endurance was normal; in flexion, they had normal strength, but averaged 20% less endurance. Testing of the group that had the nondominant extremity repaired revealed a supination strength deficit of 25%, but normal endurance. Flexion strength and endurance were essentially normal in this group. Anatomic repair of a distal biceps tendon rupture gives consistently excellent subjective and good objective results in athletes, particularly for those sports with high strength demands such as weight lifting and body building. Rehabilitation of the operated arm, especially the repaired nondominant extremity, should be emphasized. PMID- 8427352 TI - Articular cartilage lesions in ankles with lateral ligament injury. An arthroscopic study. AB - To investigate intraarticular lesions producing persistent postoperative pain, we arthroscopically examined 31 ankles in 31 patients (15 women and 16 men) with lateral ligament injury. The patients ranged in age from 15 to 33 years, with a mean of 20 years. Nine patients were freshly injured, and 22 patients had chronic injuries. All of the patients underwent arthroscopic examination immediately before the ligament operation. Chondral lesions were found in 89% of the freshly injured ankles and 95% of the ankles with chronic injuries. Most of these lesions were in the medial half of the ankle joint, especially in the anteromedial edge of the tibial plafond. After followup for 1 year postoperatively, persistent pain was noted in 4 patients who had chondral lesions of greater than one-half the thickness of the articular cartilage. Pain and tenderness were localized at the anteromedial joint line, corresponding to the location of the chondral lesions. Chondral lesions of greater than one-half the thickness of the articular cartilage were found in 8 ankles in the chronic injury group, but there were none in the fresh injury group. Thus, in lateral ligament injuries of the ankle, the longer the time elapsed from the initial injury, the more severe the associated chondral lesions became. These chondral lesions appear to cause persistent pain. PMID- 8427353 TI - Acute repair of injury to the anterior cruciate ligament. A long-term followup. AB - We reviewed 30 patients at an average of 7.4 years after acute repair of the anterior cruciate ligament augmented with a loop of iliotibial tract. A noncontact twisting had been the mechanism of injury in 18 of these patients, with 28 having been injured in sports. At followup, 25 patients had not experienced symptoms of instability and 23 were able to return to unrestricted athletic activity; only 5 had been unable or unwilling to return to sporting activity at all. There had been no swelling in 23 patients; however, 17 suffered from pain on exertion. The average Lysholm score was 93.2. Joint laxity was assessed and anteroposterior tibial translation quantified with a KT-1000 arthrometer. Eighteen patients had a normal or 1+ Lachman test and 27 had an absent or 1+ pivot shift. When compared with the results of a similar study performed on this group of patients at 2 years after surgery, there had been little subjective change in knee function. However, objectively there had been significant deterioration of the anteroposterior stability of the knees at 7 years, suggesting failure of the integrity of the repaired ligament with time. An associated medial collateral ligament injury had a significant adverse effect both on the integrity of the anterior cruciate ligament repair and the incidence of postoperative stiffness. PMID- 8427354 TI - Osteochondral lesions of the talus. AB - We report demographic, clinical, and imaging data on 92 patients with osteochondral lesions of the talus collected in one center between 1981 and 1992. All patients reported pain as their primary symptom. Ninety-four percent of the patients reported pain with activity. Physical examination was unhelpful. Using newer imaging techniques (bone scan and computed tomography) and with increased awareness, we have observed a sevenfold increase in the diagnostic frequency of osteochondral lesions of the talus between the years 1981 to 1986 and 1987 to 1992. Bone scan is an excellent screening tool for patients with chronic ankle pain and has 99% sensitivity in depicting osteochondral lesions. Computed tomography demonstrated a previously unclassified lesion, the radiolucent defect, which accounts for 77% of the lesions in this series. We have therefore modified the Berndt and Harty classification system, basing it on radiographic appearance (principally computed tomography) and adding the radiolucent lesion. Fifty-eight patients were treated surgically. Anterior and midtalar lesions are now approached arthroscopically. Surgical treatment of the radiolucent lesion, consisting of curettage and drilling, gives 42% excellent and 32% good results. Pain relief often occurs within months of surgery, but healing of the lesion requires years, and some may persist indefinitely. PMID- 8427355 TI - Nonoperatively treated isolated posterior cruciate ligament injuries. AB - To evaluate the theory that isolated posterior cruciate ligament injuries do well when treated nonoperatively, we reviewed 40 patients (mean age, 33 years at followup; average interval from injury, 6 years) who completed a modified Noyes knee questionnaire and were reevaluated by physical examination, radiographs, and isokinetic testing. Thirty of the injuries to the posterior cruciate ligament were sports-related. On the questionnaire, 65% of the patients revealed that their activity level after injury was limited and 49% stated that the involved knee had not recovered fully despite rehabilitation. Ninety percent complained of knee pain with activity and 43% complained of problems with walking. The longer the interval between injury and this followup, the lower the knee questionnaire score and the greater the radiographic degenerative changes. The patients as a group exhibited excellent muscular strength with a mean isokinetic score of 99% of the contralateral extremity. There was no correlation between isokinetic testing and knee questionnaire score. Patients with greater posterior laxity, as measured by the posterior drawer examination, appeared to have greater subjective complaints. Our study suggests that patients with isolated posterior cruciate ligament injuries treated nonoperatively may maintain excellent muscle strength, but significant symptoms and degenerative changes increase with increasing interval from injury. PMID- 8427356 TI - Incidence and nature of sports injuries in Ireland. Analysis of four types of sport. AB - The objectives of this study were to investigate the incidence of sports injuries in Ireland and to analyze various ways of quantifying the seriousness of these injuries. A 12-month, prospective study was carried out on 324 Irish athletes involved at a high level of sports participation in one of the following categories: endurance, contact, noncontact, or explosive sports. Results were expressed in four ways: 1) number of injuries per year; 2) days injured per year; 3) number of injuries per 10,000 hours of participation; and 4) duration of injury per 1000 hours of participation. The average athlete sustained 1.17 acute and 0.93 overuse injuries per year and suffered the effects of sports injury for 52 days. More time was lost through overuse injuries than acute injuries. The incidence of acute injuries per 10,000 hours of participation was lowest in the noncontact sports and highest in the contact sports, but there was no difference in the incidence of overuse injuries between any of the four categories of sport. The injury rate per 10,000 hours of participation was lowest in noncontact and explosive sports and highest in contact sports. However, when expressed in terms of days lost per 1000 hours of participation, endurance sports had the lowest incidence of time loss and explosive sports the highest. PMID- 8427357 TI - A simple surgical approach to the posteromedial ankle. PMID- 8427358 TI - Fracture of the femur after anterior cruciate ligament reconstruction with a GORE TEX prosthetic graft. A case report. PMID- 8427359 TI - Osteolysis of the distal clavicle in a female body builder. A case report. PMID- 8427360 TI - Posttraumatic changes in the posterior glenoid and labrum in a handball player. AB - Magnetic resonance imaging evaluation of a handball player with chronic pain in his nondominant shoulder revealed posttraumatic changes in the posterior glenoid and labrum. This provided a pathophysiologic explanation for the patient's symptoms. PMID- 8427361 TI - Thoracic spine fracture in a football player. A case report. AB - Compression fractures of the thoracic spine are less common than injuries to the cervical, thoracolumbar, or lumbar regions because of the inherent stability and geometry of the thoracic spinal region. This injury may be difficult to detect acutely because of the lack of localizing symptoms. An appropriate index of suspicion should be present when examining a player experiencing acute back pain associated with localized spinal tenderness. Early recognition and appropriate treatment are needed to maximize the patient's chances of return to normal function while avoiding further injury from lack of recognition of the injury or too early return to play. PMID- 8427362 TI - Fluoroscopic evaluation for subtle shoulder instability. PMID- 8427363 TI - High tibial osteotomy and ligament reconstruction in varus angulated, anterior cruciate ligament-deficient knees. A two- to seven-year follow-up study. AB - We assessed short-term treatment results of younger patients with varus malalignment and chronic anterior cruciate ligament deficiency. Forty-one patients (mean, 32 years; range, 16 to 47) underwent a high tibial osteotomy. Because of giving way symptoms, 14 also had a lateral iliotibial band extraarticular procedure at the time of the osteotomy and 16 had an intraarticular anterior cruciate ligament allograft reconstruction after the osteotomy. All returned for followup (mean, 58 months; range, 23 to 86), which included KT-1000 arthrometer testing and evaluation by our knee rating system. Statistically significant (P < 0.05) improvements were found in the mean overall rating scores for pain, swelling, and giving way. Preoperatively, 30 (73%) had pain with activities of daily living or with any sports activity; 11 (27%) could perform only light sports activities without pain. At followup, 32 patients (78%) had no pain with activities of daily living or light sports. Ten of 15 patients with advanced medial tibiofemoral arthrosis (subchondral bone exposure) had significant improvements in symptoms. Patient satisfaction was high: 88% stated they would undergo the procedure again and 78% felt their knee condition was improved. Patients who had the allograft reconstruction had significantly lower (P < 0.05) anterior-posterior displacements at followup than those who had the extraarticular procedure. We concluded that osteotomy should be performed early in the disease process for younger athletes who experience symptoms with activity. It may be unrealistic, however, to expect continuation of sports beyond light recreational, given the joint arthrosis that is usually present and the high in vivo joint loadings with athletes. Anterior cruciate ligament reconstruction should be considered when giving way previously occurred and the patient plans to resume athletics. However, patients with advanced arthrosis can avoid anterior cruciate ligament surgery by reducing athletic activities. PMID- 8427364 TI - Postoperative return of motion in anterior cruciate ligament and medial collateral ligament injuries. The effect of medial collateral ligament rupture location. AB - Twenty consecutive patients with combined anterior cruciate/medial collateral ligament injuries were analyzed to determine if a correlation exists between the location of medial collateral ligament disruption and postoperative return of motion. All patients were treated operatively by autogenous patellar tendon anterior cruciate ligament reconstruction and primary medial collateral ligament repair. The mean followup was 379 days. The patients (12 men and 8 women; mean age, 23 years) were divided into two groups based on the location of superficial medial collateral ligament rupture. Group P consisted of 13 patients with lesions at or proximal to the joint line; Group D consisted of 7 patients with disruptions distal to the joint line. Group D patients had a more rapid return of motion for both flexion and extension. At the conclusion of followup, patients from Group D also achieved 8 degrees more flexion and 3 degrees more extension. There were eight additional procedures performed on five patients, all from Group P, required to treat difficulty regaining motion. Among these patients with anterior cruciate/medial collateral ligament injuries there are two distinct groups, each with different prognoses related to return of motion based on the location of the medial collateral ligament disruption. We suggest that patients with double-ligament injuries, where the medial collateral ligament lesion is proximal, should be managed very aggressively to regain motion. PMID- 8427365 TI - An in vitro dynamic evaluation of prophylactic knee braces during lateral impact loading. AB - To determine the ability of prophylactic knee braces to reduce or limit medial collateral and anterior cruciate ligament elongation under dynamic loading conditions, we used cadaveric specimens that had a surrogate soft tissue material that matched the tissue compliance of in vivo contracted muscles. Eight cadaveric specimens were fitted with four prophylactic knee braces and instrumented with Hall Effect Strain Transducers on both the medial collateral and anterior cruciate ligament. Each specimen was mounted in a testing frame while a lateral impact was applied to the knee joint by a pendulum at levels below the injury threshold. Legs were tested at 0 degrees and 30 degrees of knee flexion, both with and without an intact anterior cruciate ligament. The maximum elongation for each ligament was calculated as a percentage of the initial measured length. The addition of a prophylactic knee brace significantly reduced the level of impact force at the point of impact, but this did not result in a significant reduction of anterior cruciate ligament elongation for any test. Although not significant, all braces tested were more effective at reducing medial collateral ligament elongation during a lateral impact with the knee flexion at 30 degrees than at 0 degrees. PMID- 8427366 TI - A comparison of anterior compartment pressures in competitive runners and cyclists. AB - Anterior compartment pressure was measured in 10 competitive runners and in 10 competitive cyclists who were asymptomatic for compartment syndrome. Pressures were measured at rest, after exercise at 80% VO2max, after maximal exercise, and 15 minutes after both exercise bouts. No difference in compartment pressure was found after exercise at 80% VO2max in runners and cyclists. Total creatinine phosphokinase enzyme levels measured before and after exercise at 80% VO2max showed a 10-fold increase in runners as compared to cyclists. Anterior compartment pressure measured after maximal exercise was significantly greater in runners as compared to cyclists. Compartment pressure showed no increase from resting values during cycling at 80% VO2max or maximal exercise. These findings suggest that patients with chronic anterior compartment syndrome may be able to cycle without elevation of compartment pressure and concomitant pain as an alternative exercise to maintain a continued degree of fitness and training. Compartment pressures should be measured during cycling in patients with chronic compartment syndrome to determine its efficacy as a method for maintenance of cardiorespiratory fitness. PMID- 8427367 TI - Effect of water running and cycling on maximum oxygen consumption and 2-mile run performance. AB - This study compared water running, cycling, and running for maintaining VO2max and 2-mile run performance over a 6-week training period. Thirty-two trained subjects between the ages of 18 and 26 were evaluated for maximum oxygen uptake (VO2max) and 2-mile run performance. Subjects were stratified by a 2-mile run pretest into high, medium, and low performance levels and then randomly assigned to water running, cycling, or running training. The three groups trained with similar frequency, duration, and intensity overa 6-week period. After 6 weeks of training, all of the groups made a small but statistically significant decrease in fitness (VO2max), but no change in 2-mile run time. However, there were no differences with respect to either training modality or pretraining performance level. It was concluded that over a 6-week period, runners who cannot run because of soft tissue injury can maintain VO2max and 2-mile run performance similar to running training with either cycling or water running. PMID- 8427368 TI - A standard measure for exercise prescription for aqua running. AB - Aqua running has been promoted as a method for cardiovascular conditioning for the injured athlete as well as for others who desire a low impact aerobic workout. Recent studies have suggested the need for an environment-specific measure of exercise intensity. Twenty healthy subjects, 10 men and 10 women, underwent a graded exercise test of aqua running to investigate the relationship between cadence and heart rate. This was done to determine the utility of cadence as a measure for exercise prescription. The graded exercise test followed a standard protocol for exercise testing in aqua running. Results demonstrated a high correlation between cadence and heart rate, both as a group as well as individually. We conclude that cadence may be used as a measure for exercise prescription for aqua running. PMID- 8427369 TI - How iliotibial tract injuries of the knee combine with acute anterior cruciate ligament tears to influence abnormal anterior tibial displacement. AB - A knowledge of the patterns of injury to the components of the iliotibial tract allows a clearer interpretation of motion limits testing in patients with abnormal anterior tibial translation of the knee (anterior cruciate ligament deficient knees). Eighty-two consecutive patients with acute knee injuries were classified as anteromedial-anterolateral rotatory instability (anterior cruciate ligament-deficient) based on the abnormal motion demonstrated by clinical examination tests for instability. At surgery, injuries to the intraarticular and extraarticular anatomic structures were identified and correlated to the abnormal grades of motion demonstrated by the knee motion limits examination. Tears of the anterior cruciate ligament occurred in 80 (98%) of the knees. However, the grade of abnormal motion demonstrated by the Lachman and pivot shift tests was quite variable. This variation did not correlate statistically with anterior cruciate ligament tears. Injuries to the anatomic components of the iliotibial tract were confirmed in 76 (93%) of the knees. These injuries correlated highly with variations in grades of abnormal motion detected by the following tests: lateral joint line opening at 30 degrees (r2 = 0.05); Lachman test (r2 = 0.08); pivot shift (r2 = 0.16); and anterior translation at 90 degrees of flexion (r2 = 0.34). Thus, injuries to the components of the iliotibial tract are thought to contribute to the variation in grades of abnormal motion observed in this complex subgroup of anterior tibial translation instabilities. PMID- 8427370 TI - The strength characteristics of internal and external rotator muscles in professional baseball pitchers. AB - The purpose of this study was to establish a data base regarding the isokinetic muscular performance characteristics of the external/internal rotator muscles of professional baseball pitchers. One hundred fifty healthy professional baseball pitchers were evaluated by use of a Biodex isokinetic dynamometer. The subjects tested had a mean age of 23.4 years and a mean body weight of 199 pounds. Isokinetic tests were performed concentrically at 180 and 300 deg/sec for both the throwing and nonthrowing shoulders. Testing procedures regarding positioning and stabilization followed established guidelines. The testing protocol and actual test repetitions were standardized for each subject. Statistical analysis was performed using the Pearson Product Moment Correlation and paired t-tests. Determination of the correlation coefficient was made at the P < 0.05 level of significance. Test results for bilateral comparison of mean peak torque for the throwing and nonthrowing shoulders indicated no statistically significant difference between the internal rotators at both test speeds, or for the external rotators at 300 deg/sec. There was a significant statistical difference at the 180 deg/sec test speed for the external rotators. The external/internal rotator strength ratio indicated a 65% ratio at 180 deg/sec and a 61% ratio at 300 deg/sec. Data were also collected for mean peak torque/body weight ratios of the throwing shoulder to establish a data base in professional throwers. This study offers clinical relevance in establishing a muscle performance profile for the professional thrower. This data base can therefore be used as criteria that should be met before an injured pitcher can be returned to throwing at the professional baseball level. PMID- 8427371 TI - A survey of interfering shoulder pain in United States competitive swimmers. AB - The prevalence of shoulder pain in United States competitive swimmers has not been extensively surveyed but is perceived as common. To evaluate this concern, a questionnaire survey was conducted on 1262 United States swimmers: 993 age group, 198 senior development, and 71 national team athletes. We sought to identify the incidence of interfering shoulder pain in this population and how it is influenced by various training tasks. The prevalence of current shoulder pain in these groups varied between 10% (age group) to 26% (national team) and increased with time in the sport. In those athletes with a painful shoulder, weight training, use of hand paddles, kickboard use, stretching, and various resistance activities aggravated the painful shoulder. This survey has identified that interfering shoulder pain is present in a substantial number of competitive swimmers. PMID- 8427372 TI - Arthroscopic resection of the acromioclavicular joint. AB - Arthroscopic resection of the distal clavicle was used to treat 26 patients who had osteoarthritis of the acromioclavicular joint. Twenty of these patients were available for review at a minimum followup of 2 years. The preoperative ratings for pain, activities of daily living, work, and sports improved markedly in 17 patients postoperatively. No intraoperative complications were noted. The operation was unsuccessful in 3 patients and all underwent a second, open surgery. The results show that arthroscopic resection was effective in the treatment of isolated acromioclavicular joint arthritis. PMID- 8427373 TI - Intercollegiate ice hockey injuries. A case for uniform definitions and reports. AB - The lack of agreement on definition of terms and consistent reporting strategies in sports epidemiology complicates the determination of injury rates in any sport. This study describes Canadian Intercollegiate ice hockey injuries over a 6 year period by following a standardized reporting strategy and clearly defined terminology. Overall, the data show that the knee is most susceptible to injury, that the forwards recorded the highest number of injuries, and that body contact caused the majority of injuries. Compared to other studies the results indicate a decreasing per game injury rate over the last 15 years and provide evidence that helmets and visors reduce the risk of head and facial injuries. Recommendations are propagated toward the adherence of standardized reporting strategies and uniform definitions to be used in future sports injury epidemiologic research. PMID- 8427374 TI - Isometric versus tension measurements. A comparison for the reconstruction of the anterior cruciate ligament. AB - This study was designed to compare the displacement patterns of an isometer, used to determine graft placement during reconstruction, with the actual tensions on an anterior cruciate ligament substitute. In cadaveric specimens, a Kevlar anterior cruciate ligament substitute was implanted in three separate femoral sites, each of which was subsequently fixed to two different tibial sites. The initial tension of the Kevlar substitute was set to 22 or 33 N at 20 degrees of knee flexion. The displacement patterns for each position were recorded during passive flexion-extension using the isometer. Using a custom-designed tensiometer, the tensile forces on the substitute after rigid fixation at the tibia and femur were measured. During passive flexion-extension, the maximum change in tension of the anterior cruciate ligament substitute, measured by the tensiometer, was correlated with the maximum change in displacement between attachment sites, measured by the isometer. The coefficient of determination was equal to 0.15, indicating that the isometer may not accurately predict the tensions developed in the substitute. PMID- 8427375 TI - Radiographic imaging of muscle strain injury. AB - We reviewed our experience with computed tomography and magnetic resonance imaging of acute muscle strain injury. We imaged 50 athletes (average age, 28 years; range, 17 to 42) who had an acute muscle strain involving either the adductor, hamstring, quadriceps, or triceps surae muscles. Computed tomography (axial imaging) was used from 1982 to 1987 for 27 athletes. Spin-echo magnetic resonance imaging (axial, coronal, sagittal imaging) was used from 1987 to 1991 for 23 athletes. Computed tomography and magnetic resonance imaging localize the strain injury to a single muscle within a group of synergists; the adductor longus, rectus femoris, and medial head of gastrocnemius muscles are most prone to strain injury. A disruption occurs predictably at the myotendinous junction; fluid collects at the disruption site and dissects along the epimysium and subcutis. Muscle tissue remote from the myotendinous junction clearly demonstrates extensive injury with abundant magnetic resonance imaging signal changes consistent with edema and inflammation. Follow-up computed tomographic and magnetic resonance imaging studies can clearly demonstrate atrophy, fibrosis, and calcium deposition. PMID- 8427376 TI - Does amateur boxing lead to chronic brain damage? A review of some recent investigations. AB - Fifty former amateur boxers were examined and compared with two control groups of soccer players and track and field athletes. All subjects were interviewed regarding their sports career, medical history, and social variables. They underwent a physical and a neurologic examination. Personality traits were investigated and related to the platelet monoamine oxidase activity. Cerebral morphologic changes were evaluated using computed tomography and magnetic resonance imaging. Further, clinical neurophysiologic tests were made as well as neuropsychologic tests. No significant differences were found between the groups in any of the physical or neurologic examinations or in platelet monoamine oxidase activity. Socially, the boxers had a lower degree of education and had chosen less intellectual professions, but they were less impulsive and more socialized. The computed tomography images and magnetic resonance imaging studies showed no significant differences between the groups. There was a significantly higher incidence of slight or moderate electroencephalography deviations among the boxers. Neuropsychologically, the boxers had an inferior finger-tapping performance. Thus, no signs of serious chronic brain damage were found among any of the groups studied. However, the electroencephalography and finger-tapping differences between the groups might indicate slight brain dysfunction in some of the amateur boxers. PMID- 8427377 TI - Studies on the control of visceral leishmaniasis: validation of the Falcon assay screening test--enzyme-linked immunosorbent assay (FAST-ELISA) for field diagnosis of canine visceral leishmaniasis. AB - The Falcon assay screening test-enzyme-linked immunosorbent assay (FAST-ELISA), the latest version of the enzyme-linked immunosorbent assay, uses antigen-coated beads. A 96-well plate can be run in 20 min without electricity or expensive equipment. We compared the FAST-ELISA, a standard ELISA, and an indirect immunofluorescent assay (IFA) for evaluation of canine leishmaniasis under field conditions using samples from 161 dogs from our longitudinal study in the endemic area of Jacobina, Bahia, Brazil. Organisms were isolated by culture (NN medium) or by inoculation of hamsters with samples from 59 of the dogs. When plasma were tested, we found a sensitivity of 88% and a specificity of 90% using the FAST ELISA with a spectrophotometer. Using the same plasma samples, the IFA had a sensitivity of 75% and a specificity of 93%. The standard ELISA had a sensitivity of 90% and a specificity of 85%. When whole blood was tested with the FAST-ELISA, we found a sensitivity of 85%. There was no significant difference between visual and spectrophotometric results with plasma or whole blood. The FAST-ELISA system provides a sensitive, specific, and field-adaptable test for canine visceral leishmaniasis, which can be evaluated quickly without the use of a microscope or spectrophotometer. PMID- 8427378 TI - Emergence of multidrug-resistant Salmonella typhi in rural southern India. AB - Multidrug-resistant Salmonella typhi has spread to many parts of India, causing severe therapeutic problems. Of the 305 clinically suspected cases of enteric fever seen at Kasturba Hospital in Manipal, Karnataka between January 1990 and June 1991, Salmonella bacteremia was detected in 134 patients; 102 of these were caused by S. typhi. Eighty (78.4%) of the isolates from S. typhi-infected patients were resistant to conventional antibiotics used in the treatment of typhoid fever (i.e., ampicillin, chloramphenicol, and trimethoprim sulfamethoxazole). No in vitro resistance was observed to ciprofloxacin, norfloxacin, and gentamicin. Major complications were seen in four patients infected by multiresistant S. typhi; three responded well to ciprofloxacin. The fourth patient died of hemorrhage and acute renal failure, even after receiving ciprofloxacin. In addition, less severe complications such as hepatitis and jaundice were observed in 12 other patients. Seventy-six of the multiresistant S. typhi belonged to phage type O biotype II and four were of untypeable Vi strains. The emergence of multidrug-resistant S. typhi has necessitated the use of fluoroquinolones in the therapy for enteric fever. PMID- 8427379 TI - Inhibitory effects of sinefungin and its cyclic analog on the multiplication of Trypanosoma cruzi isolates. AB - Sinefungin and its cyclic analog were evaluated in vitro for activity against the multiplication of Trypanosoma cruzi. When either drug was tested for eight days on twelve T. cruzi epimastigote isolates, an 800-fold difference in drug sensitivity was found. Both drugs were trypanostatics at a concentration range from 0.1 micrograms/ml to 300 micrograms/ml. The 50% effective concentration (EC50) of sinefungin and its cyclic analog at which the growth of a given isolate was inhibited was 0.38 micrograms/ml for sinefungin and 0.31 micrograms/ml for the cyclic analog against the Ma, Marin, OPS-86, Y, and Ya isolates, and > 300 micrograms/ml for sinefungin and 217 micrograms/ml for the cyclic analog against the A-35, Bertoldo, DS, EP, ES, OPS-58, and FL isolates. Incubation of drug sensitive isolates for more than 10 days in glucose-saline (GS) medium, but not in minimal essential medium, in the presence of a 30-fold EC50 concentration of the drug induced an increase in the drug-resistant population, which maintained this phenotype for several passages in drug-free culture medium. Growth curves were analyzed as a function of parasite inoculum; it was observed that with sinefungin-sensitive T. cruzi epimastigote isolates grown in GS medium in the presence of 10 micrograms/ml of the drug, the inhibitory effects of the drug were dependent on the initial inoculum: 1 x 10(3)-1 x 10(4) parasites/ml were strongly inhibited even after 16 days. Significant impairment of thymidine incorporation into the DNA of parasites by both drugs was observed only in drug-sensitive epimastigote isolates.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427380 TI - In vitro activity of monodesethylamodiaquine and amopyroquine against African isolates and clones of Plasmodium falciparum. AB - The in vitro activity of monodesethylamodiaquine and amopyroquine was evaluated against chloroquine-susceptible and chloroquine-resistant African clones of Plasmodium falciparum. The chloroquine-resistant clone (50% inhibitory concentration [IC50] = 1,630 nM) was 26 and 3.7 times less susceptible to monodesethylamodiaquine and amopyroquine, respectively, than the chloroquine susceptible clone (IC50 = 36.5 nM). Cross-resistance patterns of chloroquine, monodesethylamodiaquine, and amopyroquine were determined against 62 African isolates of P. falciparum obtained from imported cases of malaria in France, using an isotopic semimicro drug susceptibility test. Chloroquine-resistant isolates (n = 38) were significantly less susceptible to both monodesethylamodiaquine (IC50 = 74.8 nM) and amopyroquine (IC50 = 18.9 nM), as compared with the 24 chloroquine-susceptible isolates (IC50 = 28.2 nM and 16.1 nM, respectively). A significant positive correlation was found between chloroquine and monodesethylamodiaquine (r = 0.903) and between chloroquine and amopyroquine (r = 0.371), indicating in vitro cross-resistance among these drugs. These in vitro results suggest that the clinical response to amodiaquine and amopyroquine needs to be closely monitored in Africa. PMID- 8427381 TI - Reversal of Plasmodium falciparum resistance to chloroquine in Panamanian Aotus monkeys. AB - An Aotus-Plasmodium falciparum model was used to determine if chloroquine resistance could be reversed in vivo. The putative resistance modulators tested all reverse chloroquine resistance in vitro and included verapamil, chlorpromazine, prochlorperazine, cyproheptadine, ketotifen, a tiapamil analog (Ro 11-2933), and a chlorpromazine analog (SKF 2133-A). Combinations of chloroquine plus chlorpromazine or prochlorperazine confirmed reversal of chloroquine resistance as exhibited by cures obtained in six Aotus monkeys infected with chloroquine-resistant P. falciparum (Vietnam Smith/RE strain) and rapid clearance of parasitemia, followed by recrudescence in six additional monkeys. The results indicate the following order of in vivo efficacy for reversing chloroquine resistance in Aotus: chlorpromazine > prochlorperazine >> desipramine >> Ro 11-2933 (tiapamil analog) > ketotifen. Cyproheptadine and verapamil were not effective in reversing chloroquine resistance and probable drug toxicity was observed with these drugs in combination with chloroquine. PMID- 8427382 TI - Pulsed-field gel electrophoresis as a method of biotyping of Giardia duodenalis. AB - Pulsed-field gel electrophoresis (PFGE) has allowed karyotypic characterization of protozoa at a species and a subspecies level. Different methods of PFGE have been applied to analysis of Giardia isolates from the major morphologic groups and of isolates from the G. duodenalis group. We developed a four-day method of contour-clamped homogeneous electric field PFGE that produced reliable chromosomal band separation for isolates from the G. duodenalis group. When a significant waterborne outbreak of giardiasis occurred in a town in British Columbia, this biotyping method was assessed by characterization of isolates associated with the outbreak. Results were compared with karyotypes obtained from isolates retrieved in other parts of the province, as well as with reference isolates. Isolates in this study were found to have 4-6 chromosomal bands estimated to be between 1 and 4 Mb in size. The chromosomal complement of the outbreak-associated isolates was relatively homogeneous and distinguishable from the non-outbreak-associated British Columbia and reference isolates. The present study showed this PFGE method to be useful in biotyping isolates of G. duodenalis. This study also exemplifies the degree of heterogeneity displayed at the genotypic level of Giardia, as well as the apparent stability of the outbreak associated karyotype during transmission from original infecting animal through a water vehicle to human cases. PMID- 8427383 TI - A study of onchocerciasis with severe skin and eye lesions in a hyperendemic zone in the forest of southwestern Cameroon: clinical, parasitologic, and entomologic findings. AB - Prior to the initiation of an onchocerciasis control program based on the mass administration of ivermectin in the rain forest of southwestern Cameroon, a preliminary baseline study of the area was conducted. The results of this study showed that onchocerciasis was hyperendemic in the area. Skin symptoms and signs were observed including pruritus (67.4% of the population examined), onchocerca nodules (51.6%), skin depigmentation (18.5%), and hanging groins (5.7%). Except for pruritus, the prevalence of these symptoms increased with age. Of the eyes examined, 44.9% had microfilariae in the anterior chamber, 33.5% had choroidoretinitis, 28.0% had punctate keratitis, 8.3% had papillary abnormalities, and 3.6% had sclerosing keratitis. Vision in 10.5% of the eyes examined was classified as blind or very poor (visual acuity = 0-0.10), in 15.7% as poor (visual acuity = 0.11-0.39), and in 73.8% as good (visual acuity = 0.4 1.00). Unlike previous reports that have linked serious ocular damage mainly to savanna onchocerciasis, the present study showed that forest onchocerciasis also caused significant ocular pathology, including blindness. Parasitologically, positive skin snips were recorded for 92.7% of the persons examined, with both sexes being equally infected. The parasite load, expressed as the geometric mean number of microfilariae per skin snip, was 53.6, and was much higher in males than in females. The flv vector, Simulium squamosum, had a high infection rate of 7.5% infective females in Bakumba and 6.8% infective females in Ngbandi, the two fly-catching points. The transmission potential was 266 infective larvae per person per month in Bakumba and 189 in Ngbandi. PMID- 8427384 TI - Leishmania colombiensis in Venezuela. AB - Flagellate parasites isolated in Venezuela from bone marrow aspirates of a human (MHOM/VE/70/Chuao) and a dog (MCAN/VE/72/Talisman2) were subsequently identified by isozyme analysis as Leishmania colombiensis. Data are presented describing genetic similarity among Panama, Colombia, and Venezuela populations of this species. PMID- 8427385 TI - Detection of diarrheogenic Escherichia coli in children less than ten years old with and without diarrhea in New Caledonia using seven acetylaminofluorene labeled DNA probes. AB - We report the use of seven acetylaminofluorene (AAF)-labeled DNA probes in evaluating the incidence of various Escherichia coli pathotypes in New Caledonia among 448 children with acute diarrhea (1,278 E. coli pathotypes studied) and 88 controls (264 E. coli pathotypes studied) in 1990. Diarrheogenic E. coli were detected using cloned gene probes for heat-labile and heat-stable enterotoxins, Shiga-like cytotoxins (SLTI and SLTII), the cell invasion phenotype (INV), and enteropathogenic-adherence factor (EAF). Isolates were also studied using bioassays and radioactive DNA probes as reference methods. Enterotoxigenic E. coli (ETEC) were isolated from only 5.36% of the patients; E. coli with localized adherence (LA) to HEp-2 cells was much more common in patients (14.4%) than in controls (3.4%; chi 2 = 7.54, P < 0.01), but most of the E. coli with an LA pattern were members of traditional enteropathogenic E. coli (EPEC) serogroups (chi 2 = 92.95, P < 0.001). Non-enteropathogenic E. coli with an LA pattern were weakly associated with diarrheal disease (8.9%). Escherichia coli with a diffuse or an aggregative pattern did not show a significant association with infantile diarrhea. Eight EPEC serogroups were identified and the frequency of positivity for the LA pattern was 70.5%; the EAF was significantly associated with the 0119:K9 serogroup. No enteroinvasive or SLT-producing E. coli were identified. An evaluation of the AAF probes in comparison with 32P-labeled probes and conventional bioassays was made during this epidemiologic survey. The positive and negative predictive values of the ETEC probes were 0.91 and 1, respectively (overall agreement = 99.8%).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427386 TI - Active surveillance and risk factors for leptospirosis in Hawaii. AB - A clinic-hospital-based leptospirosis surveillance program was conducted to determine the morbidity and risk factors in nonepidemic settings. The study was conducted on two islands, Kauai and Hawaii (Big Island), in the state of Hawaii for one year during 1988 and 1989. An active, more comprehensive case detection system was used on the Big Island that enabled us to determine the incidence of clinical disease. Subjects from both islands were used to conduct a case-control study for risk factors. One hundred seventy-two subjects from the Big Island (who presented with any two of the following symptoms: fever, headache, myalgia, or nausea/vomiting) were enrolled in the study. Twenty cases were diagnosed by culture, serology, or fluorescent antibody tissue staining at autopsy. Six cases required hospitalization and two succumbed to fatal infections. We estimated that these cases represented an annual incidence rate of 128 per 100,000 person-years in our target population. For 33 cases, 77 controls were matched for island, age, sex, and time of onset of illness. Interviews were conducted retrospectively in a double-blinded fashion with cases and controls and evaluated approximately 30 risk factors. Factors that were associated most strongly with development of leptospirosis were household use of rainwater catchment systems (P = 0.003), presence of skin cuts during the incubation period (P = 0.008), contact with cattle or the urine of cattle (P = 0.05 and P = 0.03, respectively), and handling of animal tissues (P = 0.005).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427387 TI - Prevalence of cutaneous leishmaniasis along the Nile River north of Khartoum (Sudan) in the aftermath of an epidemic in 1985. AB - Based on a pilot clinical study of the prevalence of cutaneous leishmaniasis (CL) among school children in villages on both banks of the Nile River north of Khartoum, Sudan in the aftermath of a 1985 epidemic, we studied a random sample (303 individuals) from one of these villages to determine the prevalence of infection and exposure to CL. Four percent of the population had active CL lesions, 47% had healed lesions, and another 43% reacted positively to sensitization with leishmanin in the absence of past or active CL lesions. Ninety one percent of the total population reacted positively to leishmanin. The present status of CL in the area and possible reasons for the emergence of the epidemic are discussed, and gaps in our knowledge about the epidemic are identified. PMID- 8427389 TI - Application of the polymerase chain reaction to the epidemiology of pathogenic and nonpathogenic Entamoeba histolytica. AB - We used the polymerase chain reaction (PCR) to study the epidemiology of pathogenic and nonpathogenic Entamoeba histolytica in a rural community in Mexico. Formalin-fixed stool samples were used for extraction of DNA. The PCR amplifications were performed using two sets of primers that discriminate between pathogenic or non-pathogenic E. histolytica. A total of 201 randomly selected individuals were studied. Among them, 25 (12%) were diagnosed to be infected with E. histolytica by microscopy; PCR identified 24 of these as positive (sensitivity = 0.96) and of 176 negative individuals, only three were identified as positive (specificity = 0.98). The PCR analysis defined three populations: 14 cases were positive for both pathogenic and nonpathogenic E. histolytica, nine cases were positive for pathogenic and negative for nonpathogenic E. histolytica, and only one case was negative for pathogenic and positive for nonpathogenic E. histolytica. Infection by E. histolytica was strongly associated to infection with Entamoeba coli (odds ratio [OR] = 9.41, 95% confidence interval [CI] = 3.09, 28.65, P < 0.0004) and Endolimax nana (OR = 6.15, 95% CI = 2.03, 18.17, P < 0.0004). This new technique has high specificity and sensitivity; it is simple, reproducible, fast, avoids the need to culture trophozoites, and can be applied in the field for epidemiologic studies. PMID- 8427388 TI - Entomologic and demographic correlates of anti-tick saliva antibody in a prospective study of tick bite subjects in Westchester County, New York. AB - We measured anti-tick saliva antibody (ATSA) by enzyme-linked immunosorbent assay using whole sonicated Ixodes dammini salivary glands as antigen in subjects with 1) a recent and confirmed I. dammini (n = 100) or Dermacentor variabilis bite (n = 3), 2) erythema migrans (n = 15), 3) late-stage Lyme disease (n = 4), and 4) normal controls without a history of tick bites (n = 5). Tick bite subjects had three ATSA determinations over approximately six weeks. On the first ATSA measurement at a mean +/- SD of 18.5 +/- 19.8 hr after removal of the tick, the subjects bitten by I. dammini had a mean ATSA optical density value (95% confidence interval [CI]) of 0.264 (0.223, 0.305); the corresponding value in controls was 0.142 (0.115, 0.169). There was no consistent change in ATSA levels in individuals with time. Multiple linear regression indicated that tick engorgement (P < 0.01), subject age (higher ATSA with increasing age; P = 0.01), and subject sex (females > males; P = 0.03) were all independent predictors of ATSA levels. Logistic regression revealed that a bite by I. dammini that became engorged (defined as an engorgement index > or = 3.4) was a risk factor for ATSA seropositivity (odds ratio [95% CI] = 6.2 [1.7, 21.8]). Finally, the ATSA test had a sensitivity of 0.81 and a specificity of 0.56 for a bite by I. dammini that became engorged. Overall, the data are further evidence that ATSA is a biologic marker of tick exposure, in that the engorgement index, a surrogate for tick saliva dose, was the strongest independent predictor of antibody response. PMID- 8427390 TI - Surgical extraction of guinea worm: disability reduction and contribution to disease control. AB - Surgical extraction of Guinea worm prior to eruption through the skin has long been performed by traditional healers in India. Using modern aseptic techniques under local anesthesia, unerupted worms can be completely and painlessly removed in several minutes. As a result, the average number of working days lost due to a single worm is reduced from three weeks or more to three days. In the field, the procedure results not only in a dramatic decrease in Guinea worm associated disability, but also in an improvement in detecting cases, and appears to reduce disease transmission. PMID- 8427391 TI - Liver sonography in an area endemic for schistosomiasis haematobium. AB - Through the use of portable ultrasonography, eight cases (2%) of periportal fibrosis were identified in a random sample of 400 subjects selected from a village with a high prevalence and morbidity due to schistosomiasis haematobium in the White Nile Province of Sudan. In contrast, 36 cases (15%) of fibrosis were seen in an area with a similar prevalence and morbidity due to schistosomiasis mansoni in the Gezira Managil region of Sudan. Although there was only one case of Schistosoma mansoni as determined by repeated stool examination of the entire sample population in the first village, the majority of those with fibrosis and age-matched controls showed serologic evidence of active S. mansoni infection. This led to the conclusion that the cases of periportal fibrosis seen in the White Nile Province are most probably due to S. mansoni rather than to S. haematobium. PMID- 8427392 TI - Snakebite-induced acute renal failure: an experimental model. AB - Acute renal failure (ARF) has been frequently reported after bites from the Viperidae snake family. The lack of a reproducible animal model has hampered the study of renal damage mechanisms. Intravenous injection of rats with 0.4 mg/kg of Bothrops jararaca venom produced functional and morphologic changes similar to those observed in human snakebite-induced ARF. There was an acute and significant decrease in the glomerular filtration rate, diuresis, and renal plasma flow. Serum fibrinogen levels decreased significantly. There was intravascular hemolysis, as shown by a significant decrease in hematocrit, and an increase in plasma lactate dehydrogenase levels and free hemoglobin. Blood pressure and serum creatine phosphokinase levels were not affected. Light and electron microscopy showed massive fibrin deposition in glomerular capillaries, proximal and distal tubular necrosis, and hemolyzed red blood cell casts in renal tubules. Based on these findings, a model of snakebite-induced ARF was achieved. Ischemia related to glomerular coagulation and intravascular hemolysis were the most important pathogenic factors causing a decrease in the glomerular filtration rate, although a direct venom nephrotoxicity cannot be excluded. PMID- 8427393 TI - Vitamin A status of children in the urban slums of Karachi, Pakistan, assessed by clinical, dietary, and biochemical methods. AB - We assessed the vitamin A status of 532 children with an age range of 6-60 months who were living in slum areas of Karachi, Pakistan, using three methodologies: clinical eye examination, dietary vitamin A intake, and serum retinol level. No definite clinical signs of xerophthalmia were observed in any of these children. The mean +/- SD vitamin A intake estimated from a food frequency questionnaire for the group with inadequate (low and deficient) serum retinol levels (< 20 micrograms/dl) was 362 +/- 332 retinol equivalents (RE) compared with 431 +/- 332) RE in the group with adequate serum levels (P < 0.005). Deficient serum retinol levels (< 10 micrograms/dl) were present in 12 children (2%); two of these had a healed corneal scar. Low serum retinol levels (10-19 micrograms/dl) were present in 46%, while 51% children had adequate levels (> or = 20 micrograms/dl). The mean +/- SD serum retinol level for the inadequate (< 20 micrograms/dl) and adequate groups were 15.3 +/- 2.8 and 26.6 +/- 6.7 micrograms/dl, respectively. These results suggest that a significant number of children in these communities have low vitamin A levels and thus may constitute an at risk group. These results also suggest that the dietary intake method may be a simple and inexpensive screening tool for assessment of vitamin A status in communities. PMID- 8427394 TI - A community-based trial of ivermectin for onchocerciasis control in the forest of southwestern Cameroon: clinical and parasitologic findings after three treatments. AB - In the onchocerciasis-endemic rain forest area of the Rumpi Hills in southwestern Cameroon, a community-based trial of ivermectin, given either once or twice a year over a three-year period (1988-1991), confirmed that the drug is a potent microfilaricide. The side effects recorded following the first treatment were edema, fever, pruritus, generalized body pains and lymphadenitis. Following subsequent treatments, the same adverse reactions were recorded, but these were generally milder when compared with those of the first treatment. The prevalence of skin microfilaria (mf) was more reduced in zone two, in which treatment was given every six months (76.9% reduction at the end of one year) than in the zone one, in which treatment was given once a year (7.4% reduction). In both zones, the impact of the drug in reducing the intensity of infection was more significant than that for prevalence. Besides pruritus, other skin symptoms were not significantly modified by ivermectin treatment. Ivermectin reduced the prevalence of ocular mf as well as the mf load of the anterior chamber of the eye, resulting in improvement of certain eye lesions such as punctate keratitis, anterior uveitis, and papillary anomalies. There was also some improvement in visual acuity. The level of participation of the village populations was somewhat low, ranging from 52% to 66%, despite excitement over the drug's additional benefit of expelling intestinal round worms. PMID- 8427395 TI - Meningitis and encephalitis at the Abbassia Fever Hospital, Cairo, Egypt, from 1966 to 1989. AB - A total of 7,809 patients with meningitis or encephalitis were admitted to the Abbassia Fever Hospital in Cairo, Egypt from November 1, 1966 to April 30, 1989. The etiology was Neisseria meningitidis (mostly group A) in 27.3% of the patients, Mycobacterium tuberculosis in 19.7%, Streptococcus pneumoniae in 7.3%, and Haemophilus influenzae in 4.1%. Almost 27% of the cases had purulent meningitis but without detectable etiology; however, the epidemiologic data suggest that most of these had meningococcal meningitis. Encephalitis was suspected in 12.5% of the patients. Most of the meningococcal, pneumococcal, and Haemophilus cases occurred during the winter months. The number of meningococcal and culture-negative purulent cases per year reached a maximum three times during the 22.5 years of this study. There were more males than females in all etiologic groups, with the ratio for the total patient population being 1.6:1. The average age ranged between 11.7 and 16.5 years for all groups except for Haemophilus patients, who had a mean age of 2.5 years. The mortality rate was almost 55% for tuberculous patients and was approximately 40% for both pneumococcal and Haemophilus patients; it was 8.5% in patients with meningococcal disease. PMID- 8427396 TI - Continuous infusion of cefazolin is superior to intermittent dosing in decreasing infection after hemorrhagic shock. AB - Standard doses of antibiotic administered by intermittent infusions after hemorrhagic shock have decreased efficacy in combating infection. This study compared identical quantities of cefazolin administered after shock as intermittent doses or as continuous infusions in a subcutaneous abscess model. One hour after resuscitation from shock, rats were inoculated with 2 x 10(8) Staphylococcus aureus subcutaneously on the dorsum and divided into three groups: (1) control rats, which received no drug treatment; (2) rats in the intermittent group, which received cefazolin at either 30 or 60 mg/kg intraperitoneally, 30 minutes prior to inoculation, then every 8 hours for three doses, and (3) rats in the continuous infusion group, which received cefazolin at either 30 or 60 mg/kg intraperitoneally, 30 minutes prior to inoculation, followed by cefazolin, 90 or 180 mg/kg, intraperitoneally by continuous infusion more than 24 hours after inoculation. Seven days after the inoculation, abscess number, diameter, and weight were measured. Rats that received either dosage of cefazolin intermittently had the same abscess rate after shock as control rats. Rats that received a continuous infusion of cefazolin at either dose had 56% fewer abscesses than control rats. Abscess diameter and weight decreased with increasing quantities of cefazolin, and abscesses were always smaller in rats receiving the continuous infusion. There were no differences in peak subcutaneous cefazolin levels between the intermittent and continuous groups. Continuous infusion provided significantly more cefazolin to the tissue than an equivalent quantity of cefazolin delivered as intermittent doses. These data demonstrate that continuous infusion of cefazolin provided more antibiotic to the tissue and was superior to intermittent injection in reducing infection after hemorrhagic shock. PMID- 8427397 TI - Changing clinical picture of mammary hamartoma. AB - During a 6-year period, we identified 12 patients (age range: 16 to 72 years) with a histologic or mammographic diagnosis of mammary hamartoma. The lesion was found in 9 of 441 open breast biopsy specimens (2%) and was identified radiographically in 5 of 8,122 mammographic examinations (less than 1%). Two groups of patients were identified. Three patients under 30 years of age underwent the excision of small palpable lesions found on pathologic examination to be mammary hamartomas (group I). In nine patients over age 30, masses were identified or confirmed on mammography (group II). Five lesions showed the classic mammographic appearance of a mammary hamartoma (a circumscribed tumor of mixed soft tissue and fatty density), and the other four were indeterminate. Presentation in these older women who had a relatively high incidence of atypical mammographic findings mandates that biopsy be performed. PMID- 8427398 TI - Determination of placental ferritin-positive peripheral lymphocytes in early stages of breast cancer. AB - "Ferritin-blocked lymphocytes" or placental ferritin (PLF) -positive T cells have repeatedly been described in the circulation of patients with female breast cancer. Since a monoclonal antibody directed against PLF became available, a study was performed to evaluate its usefulness in an easily reproducible system. One hundred patients with controversial or highly suspicious findings on mammography who subsequently underwent operation entered this trial. Sixty-one healthy blood donors served as controls. Patients with early (lymph-node negative) stages of breast cancer (in situ and T1N0 tumors) revealed significantly higher numbers of PLF-positive cells (9.00% +/- 4.5% and 6.21% +/- 3.4%) as compared with controls or patients with benign lumps (p < 0.001). Patients with negative lymph nodes differed significantly from node-positive patients (9.79% versus 2.55%; p < 0.001), whereas no difference as related to menopausal and estrogen-receptor status was observed. In order to define the sensitivity and specificity of this test, we analyzed four different cutoff levels (3%, 4%, 5%, and 6% of PLF-positive T cells). At a level of PLF-positive lymphocyte cells of 4%, 94% of cancer patients with stage T1N0 disease or ductal carcinoma in situ, 5% of patients with benign lumps, and 7% of healthy controls were identified. Furthermore, 88% of all lymph node-negative cancer patients had more than 4% positive cells, compared with only 25% in patients with axillary node involvement. The fact that more than 90% of all patients with in situ carcinomas and patients with stage T1N0 cancer had values above 4% offers promising aspects for this method to be used to complement mammography in the early detection of breast cancer. PMID- 8427399 TI - Impact of method of anesthesia on the accuracy of needle-localized breast biopsies. AB - Needle-localized breast biopsy (NLBB) is now a commonly performed procedure. We reviewed a series of 260 NLBBs to evaluate the accuracy of the procedure, using specimen radiography as the standard. General anesthesia was used in 114 (44%) procedures and local anesthesia in 146 (56%). The accuracy rate when the patient received general anesthesia was 96.5% compared with 98% when local anesthesia was used (not significant). The cost of this procedure can be reduced substantially by utilizing local anesthesia as opposed to general anesthesia without compromising the accuracy. PMID- 8427400 TI - Value of preoperative bone and liver scans and alkaline phosphatase in the evaluation of breast cancer patients. AB - We reviewed the results of 133 bone scans and 63 liver scans (computed tomography, liver-spleen radionuclide scan, or ultrasonography) obtained for the preoperative evaluation of breast cancer patients. Information on the preoperative staging of breast cancer (TNM classification) was available in 131 of 133 patients. Bone scans had a low preoperative yield as only 4 of 133 patients (3%) had a positive bone scan that correlated with the results of plain films. Only 1 of 63 patients had a liver scan suggestive of possible metastasis. We also found that the alkaline phosphatase level was not a good predictor of bone or liver metastases in breast cancer patients. In 103 patients with normal bone scans, the majority (54%) had elevated alkaline phosphatase levels; conversely, 9 of 30 patients (30%) with abnormal scans had normal alkaline phosphatase levels. Furthermore, 51 of 63 patients (81%) with elevated alkaline phosphatase levels had normal liver scans. Approximately $74,000 was spent on these liver and bone scans. PMID- 8427401 TI - Influence of level and depth on recurrence rate in thin melanomas. AB - A population-based study of the biology of the thin-level melanoma according to site, Breslow's thickness, and Clark's level was undertaken. Two hundred fifteen patients were studied with a mean follow-up of 41 months. Overall, 23 patients (10.7%) had recurrences, 8 locally, 9 regionally, and 6 systemically, despite an adequate local excision. A multivariate analysis was done. In the patients with thin lesions (less than 1 mm), increasing level (p < 0.002) and head and neck site (p < 0.04) increased the risk of recurrence. Increasing thickness of melanoma up to 1 mm did not influence the risk. This study identifies a group of high-risk melanoma patients for whom adjuvant therapy to decrease recurrences should be studied. PMID- 8427402 TI - Dose dependency and wound healing aspects of the use of tissue plasminogen activator in the prevention of intra-abdominal adhesions. AB - Intra-abdominal adhesions have been shown to result from the impairment of peritoneal fibrinolysis by inhibitors present in ischemic tissue. A reproducible model for the formation of intra-abdominal adhesions was utilized for the evaluation of the effectiveness of intraperitoneal applications of recombinant tissue plasminogen activator (rtPA) in adhesion prevention. Concentrations of rtPA required to overcome the inhibition of fibrinolysis in this model were estimated by titration of that amount of rtPA required to lyse blood clot in the presence of a measured amount of ischemic tissue. Adhesions were graded, and the hydroxyproline content of the abdominal wounds was analyzed. The effect of intraperitoneal administration of rtPA on adhesion formation was strongly dose related. Levels of rtPA of 0.01 mg/mL showed no effect (p < 0.75) on adhesion formation, whereas levels of 0.1 mg/mL either prevented or significantly modified the formation of intra-abdominal adhesions (p < 0.05). Concomitantly, wound hydroxyproline content was significantly reduced (p = 0.004). Prior investigations have shown a strong correlation between wound bursting strength and hydroxyproline content. The results of this study indicated that the levels of rtPA required to alter or prevent intra-abdominal adhesion formation also produce a significant impairment of the early phase of wound healing as measured by the wound content of hydroxyproline. PMID- 8427403 TI - Prognostic evaluation of perineural invasion in rectal cancer. AB - To evaluate whether perineural invasion (PNI) is an important prognostic factor in patients with rectal cancer, we reviewed the records of 373 patients who underwent curative surgery. Thirty-seven patients (9.9%) were identified as having tumors with PNI. The incidence of PNI was significantly increased in lesions categorized as stage III by the International Union Against Cancer (UICC) system (20%). There was a significant difference in local recurrence between patients with stage III lesions with PNI and those with stage III lesions without PNI (p < 0.005). Also, patients with PNI of stage III lesions had a significantly lower 8-year survival rate (26.7%, p < 0.001). We conclude that PNI is an important factor influencing the prognosis of patients with stage III disease. PNI should be classified as a subgroup of the clinical stage. PMID- 8427404 TI - Surgical management and long-term follow-up of patients with choledochal cysts. AB - The medical records of 46 patients treated for choledochal cysts at Kyushu University from 1965 to 1990 were reviewed, and long-term follow-up results were evaluated based on the type of cyst and on the choice of surgical procedure. Of 46 patients, 41 (89%) were female and 5 (11%) were male, with a mean age of 24 years at the time of the initial operation. Seventy-eight percent of patients presented with an abdominal pain, 43% with jaundice, and 33% with an abdominal mass. Only seven patients (15%) presented with the classic triad. According to the Todani classification system, 26 patients (57%) had type I cysts, 2 (4%) had type II, and 18 (39%) had type IV. Four patients (9%) had biliary tract carcinoma. At the time of the initial operation, the major associated diseases were cholangitis in 15% and choledocholithiasis in 26% of cases. In 24 patients who had undergone previous cyst enterostomy, cholangitis developed in 88%, choledocholithiasis in 25%, and hepatolithiasis in 33% of patients, indicating a high complication rate after cyst enterostomy. Seventy percent of these patients needed reoperation. In contrast, cyst excision with Roux-en-Y hepaticojejunostomy gave excellent long-term results. Thus, cyst excision with hepaticojejunostomy is the definitive treatment of choice for type I and IV choledochal cysts. PMID- 8427405 TI - Pancreatic insulinomas. AB - We report a series of 30 patients with pancreatic insulinoma treated from 1967 to 1990. Twenty-nine patients underwent surgery. In 24 patients, the lesion was a benign adenoma. The pancreatic lesion was localized preoperatively in 59% of cases (94% since 1980), and all lesions that were identifiable histologically were palpable intraoperatively. Endoscopic pancreatic ultrasonography, performed twice, appeared to be a very promising method of investigation. In the 24 patients with adenoma, 14 enucleations and 10 pancreatic resections were performed, with the enucleation rate increasing over time. One patient died during the postoperative period. Pancreatic fistulas (43%) were the most common cause of morbidity and were more common after enucleation (57% versus 29% after pancreatectomy). The mean follow-up period was 7 years. Excluding the patients with adenocarcinomas, the recovery rate was 92% (23 of 25 among whom 2 patients had transitory recurrent hypoglycemia), 2 patients who underwent corporeo-caudal pancreatectomy being diabetic (8%). PMID- 8427406 TI - Major long-term complications in 1,422 permanent venous access devices. AB - A retrospective review was undertaken of 1,422 permanent venous access devices (PVADs) implanted from 1989 to 1991 at Hahnemann University Hospital. This included 730 single-lumen Hickman catheters, 368 double-lumen Hickman catheters, 307 single-lumen Portacath infusion ports, and 17 double-lumen Portacath infusion ports. Indications for placement were as follows: antibiotics in 28%; chemotherapy in 51%; hyperalimentation in 4%; intravenous fluids in 4%; hemodialysis in 3%; and undocumented indications in 10%. There were 60 PVADs removed and/or replaced prior to the completion of intended therapy (4% overall). Indications for removal were catheter infection in 1% of cases and catheter malfunction in 3% of cases. The percentage of Portacath infusion ports removed was significantly greater than the percentage of Hickman catheters that were removed (p < 0.001). However, there was no significant relationship between catheter infection or the malfunction rate, and the number of lumens, initial indication for placement, or number of catheters placed. Life-threatening complications associated with PVAD insertion occurred in fewer than 1% of cases. The insertion of PVADs is a safe and efficient mode of long-term venous access. PMID- 8427407 TI - How American surgeons introduced radiology into U.S. medicine. AB - In 1985, Wilhelm Roentgen of Germany discovered "x-rays." His findings were so startling and the expectations for x-rays were of such inestimable value that, within just a few weeks, an English language translation of his paper was published verbatim in the United States. It was immediately apparent that the usefulness of x-rays for clinical diagnosis was implicit in their ability to demonstrate pathologic problems of the skeleton and to visualize foreign objects in the body. Not unexpectedly, it was surgeons who were the first group of clinicians to put the new discovery to immediate use. Consequently, during 1896, American surgeons were at the forefront of understanding and utilizing this new modality. By reviewing the work of William Keen, James White, De Forest Willard, and Carl Beck, it becomes apparent that U.S. surgeons acted as dutiful midwives during the birth of modern radiology. PMID- 8427408 TI - Human Genome Project. AB - The Human Genome Project is an international effort to clone and sequence the entire human genome. This audacious undertaking, estimated to cost 200 million dollars per year and require 15 years to complete, promises to be one of the most revolutionary and captivating scientific endeavors ever conceived by mankind. By knowing the sequence of the estimated 3 billion base pairs of the haploid human genome and its more than 30,000 genes, many questions will be answered. Moreover, our ability to intervene at the genetic level will be possible. This review outlines the scientific goals and methods of this project and discusses some of its ethical, legal, and social ramifications. PMID- 8427409 TI - Technique of intraluminal biliary ultrasonography during laparoscopic cholecystectomy. AB - Intraluminal ultrasonography of the common duct was performed in nine patients undergoing laparoscopic cholecystectomy, using a system comprising a 20-MHz crystal in a 95-cm, blunt-tipped 6F sheath, mechanically rotated at 1,800 rpm. The probe was introduced through an incision in the cystic duct and passed into the duodenum. When the catheter was withdrawn, excellent visualization of the common and cystic ducts and lower end of the common hepatic duct was achieved. In seven patients, the biliary tree was normal. A small calculus was discovered in the common duct in one patient. This stone was not seen on a subsequent cholangiogram and was subsequently retrieved. An additional patient had mucus or sludge noted in the duct, which cast no acoustic shadow and thus was distinguished from calculi. The technique was fast, efficient, and easy to perform in this small group of patients and holds promise for screening the common duct pathology during laparoscopic cholecystectomy. PMID- 8427410 TI - A method for safe pancreaticojejunostomy. AB - We devised a new technique to increase the safety of pancreaticojejunostomy in patients with an extended operation for pancreatic cancer. This new pancreaticojejunostomy was created by end-to-side anastomosis with four layers about 7 cm distal to the jejunal stump. The cut surface of the pancreas was placed on the seromuscular coat of the ventral aspect of the jejunum to cover the posterior surface of the anastomosis, and the anastomosis between the pancreas and the jejunum was created using fibrin glue. The pancreatic duct was intubated with a silicone tube, and its stenting tube was brought out through a opening in the jejunum. The anterior surface of the pancreaticojejunostomy was covered by the proximal jejunum as a serosal patch. We used this technique in seven patients. No patient developed an anastomotic leak or any other complication. The anastomosis is covered by the jejunum and is not open to the peritoneum. This new technique of pancreaticojejunostomy may reduce the risk of pancreatic leak, especially when an extended operation is performed. PMID- 8427411 TI - A reliable operative procedure for preparing a sufficiently nourished gastric tube for esophageal reconstruction. AB - We developed a reliable procedure for obtaining sufficient blood flow at the anastomotic site of the gastric tube for esophageal reconstruction. By utilizing the junction between the left gastroepiploic and short gastric vessels via the splenic hilar vascular arcade, the distal portion of the gastric tube could be sufficiently nourished. The application of this technique resulted in a complete prevention of postoperative anastomotic leakage after antesternal esophageal reconstruction. PMID- 8427412 TI - An "open-closed" technique for the treatment of necrotizing pancreatitis. AB - Open packing or closed continuous irrigation of the lesser sac are used for the debridement of acute necrotizing pancreatitis. Multiple explorations are often required with either technique, and morbidity remains high. The authors describe a new method of treatment combining laparostoma and continuous lavage. This technique permits continuous atraumatic debridement of necrotic tissues with closed continuous lavage while maintaining a readily available laparostoma for further exploration. The exposure and debridement of the necrotic pancreatic bed and isolation of the bowel, with construction of a closed continuous irrigation system and laparostoma, are described in detail. PMID- 8427413 TI - Thoracic outlet syndrome: a useful exercise treatment option. AB - The current study is a prospective evaluation of a supervised physiotherapy program of graduated resisted shoulder elevation exercises in eight patients with thoracic outlet syndrome. All had severe neurovascular symptoms and limited neck movements before treatment. After 3 weeks of intervention, symptoms had significantly improved in all patients, and all achieved a full range of cervical neck and shoulder movement. This study confirms the efficacy of a simple treatment program for patients with thoracic outlet syndrome. PMID- 8427414 TI - Management of the leaking rectal stump after Hartmann's procedure. AB - Hartmann's procedure is often selected in situations in which the construction of a colorectal anastomosis is considered unsafe. Paradoxically, the creation of the rectal stump involves placement of an intestinal suture line that is prone to leakage. We report our experience with 11 patients with a clinically significant leakage from the rectal stump after Hartmann's procedure, which was manifested as persistent postoperative intra-abdominal infection. A method for the treatment and prevention of this potentially lethal complication is presented. PMID- 8427415 TI - Biology and management of the midgut carcinoid. AB - Midgut carcinoid tumors derive from gut entoderm. These tumors may cause a complex of symptoms comprising the carcinoid syndrome by secreting a wide variety of bioactive agents in addition to serotonin. Such symptoms generally follow metastases to the liver but may also occur in primary ovarian or retroperitoneal tumors. After localization and biochemical characterization, the bioactivity of these tumors should be blocked by octreotide, sometimes in combination with other pharmacologic antagonists, so that primary resection may be performed safely. If curative resection is impossible, then a cytoreductive management scheme should be employed that includes surgical debulking and hepatic arterial embolization, followed by palliation with octreotide. PMID- 8427416 TI - Demonstration of acrosomal membranes using the hypo-osmotic swelling test. AB - The state of the acrosomal membranes in human spermatozoa was studied by means of the hypo-osmotic swelling test and indirect immunofluorescence using anti-boar outer acrosomal membrane antibodies. The swelling phenomenon observed in the acrosomal region was characterized by expansion of the plasma membrane without modification of the outer acrosomal membrane. PMID- 8427417 TI - Changes in the lectin binding sites on the testicular, epididymal, vas, and ejaculated spermatozoon surface of dog. AB - Changes in the localization of sperm surface glycocomponents of testicular, epididymal, vas deferens, and ejaculated spermatozoa of dog (Canis domesticus) were studied employing fluorescein isothiocyanate conjugated lectins viz., Concanavalin A (ConA), Triticum vulgaris (WGA), Maclura pomifera (MPA), and Arachis hypogaea (PNA) agglutinins. The plasma membrane clothing the acrosome of the testicular, epididymal, and vas deferens spermatozoa shows reactivity with all the lectins used. However, in the ejaculated spermatozoa, the entire sperm surface shows reactive sites for ConA, WGA, and PNA. Variation in the labelling of the cytoplasmic droplet in different stages of spermatozoon transit in the epididymis has been discussed. PMID- 8427418 TI - Effect of calmodulin-like protein from buffalo (Bubalus bubalis) seminal plasma on Ca2+, Mg(2+)-ATPase of purified plasma membrane of buffalo spermatozoa. AB - Buffalo sperm heads and tails were cleaved by sonication and isolated in relatively pure proportions i.e. 95% and 98% respectively, by discontinuous sucrose density-gradient centrifugation. Purified plasma membranes from the isolated sperm heads and tails were obtained by hypotonic treatment and brief sonication followed by discontinuous sucrose density-gradient centrifugation. Ca2+, Mg(2+)-ATPase activity was evident in plasma membrane from sperm heads and tails, although activity was greater in the latter. A calmodulin-like protein isolated from buffalo seminal plasma increased the Ca2+, Mg(2+)-ATPase of plasma membrane from the sperm heads and tails by 128 and 136% respectively. Based upon the data obtained here and elsewhere (Sidhu & Guraya, 1989a) a model is proposed which explains regulation of Ca2+ in buffalo spermatozoa and implicates calmodulin-like protein and Ca2+, Mg(2+)-ATPase in sperm acrosome reaction. PMID- 8427419 TI - Histophysiological and morphometric studies of the post-natal development of rat vas deferens. AB - The post-natal growth and differentiation of the vas deferens was studied in rats aged 1-180 d by light, and with transmission electron microscopy in parallel with a radioimmunoassay of serum testosterone. During the first week and the major part of the second week of the post-natal life the vas deferens presented embryological features. Differentiation of the proximal and distal segments of the vas started 12 d after birth. At 25 d, the epithelium of the mucosa in the distal segment already showed some adult histologic characteristics. The weight notably increased between 1 and 45 d and the higher rates of growth of their histologic components occurred until 45 or 60 d, in parallel to testosterone serum levels. These results suggest that growth and differentiation of the vas deferens are androgen-dependent events and precede the end of the first spermatogenic cycle, assuring the viability of spermatozoa that will enter the duct. PMID- 8427420 TI - Lack of immunization after intraperitoneal insemination of spermatozoa. AB - Twenty infertile patients with normal tubal patency were inseminated intraperitoneally (11 once, seven twice, and two three times) with spermatozoa (mean 14 x 10(6), range 0.6-48 x 10(6)) prepared by the standard swim-up technique. The occurrence of immunization to spermatozoa was looked for by the Gelatin Agglutination Test (GAT) and Tray Agglutination Test (TAT). Both tests gave negative results for all the controls (10 pregnant and 10 puerperal women). Antisperm antibodies were measured in the serum before, 30 d and 4-7 months after Intraperitoneal Insemination (IPI). The last check was done for only 14 patients, since six became pregnant as a consequence of the first treatment. Of the 14 patients studied after 4-7 months, seven had two, and two had three IPI. In the group of inseminated patients, 18 women with no basal sperm antibody did not show evidence of antibody formation after the treatment and it was not increased after insemination in the two patients who already had low antibody titre (1/32). In conclusion, despite the large number of spermatozoa inseminated and even after several IPI attempts, there was no evidence of de novo production of or increase in already present anti-sperm antibodies according to the methods used for the detection of ASA in this study. PMID- 8427421 TI - Effects of cyproterone acetate on the ultrastructure of the human epididymis. AB - The effects of cyproterone acetate (CPA), an antiandrogenic steroid, on the ultrastructure of the epithelial cells of the human epididymis were studied. Alterations in the cell size, in the cytoplasmic and surface characteristics, and in the morphology of different organelles are described. The different ultrastructural changes observed suggest that the absorptive and secretory functions of the principal cells are impaired following the CPA treatment. By contrast, with the drastic response of the principal cells of the ductuli efferentes and caput epididymidis, the epithelial cells of the cauda epididymidis appeared less affected. The suggestion of a differential androgen dependence among the different regions of this androgen target organ, as well as between the major cell-types of the epididymal epithelium, is briefly discussed. PMID- 8427422 TI - Assessment of Palomo's operative method for infertile varicocele. AB - Palomo's operative method was employed in the surgical treatment of 58 cases with infertile varicocele, and the operative method and therapeutic results were assessed. With the lapse of time after operation, statistically significant improvement was achieved in the sperm density and sperm motility. The post operative impregnation rate was 55.2%. When the pre-operative sperm motility index had been 20% or less, the post-operative impregnation rate was 23%, which was significantly lower than the overall rate of 55.2%. Thus, the pre-operative sperm motility index is a good indicator of the post-operative capacity for impregnation. Even in patients with severe oligozoospermia (< 5 x 10(6) ml-1), a post-operative impregnation rate of 45.5% was obtained. Thus, their prognosis is by no means hopeless. Study of the cross-sectional profile of the internal spermatic vessels revealed the possibility of overlooking small internal spermatic venous branches. Therefore, Palomo's method was concluded to be excellent as an approach performed under direct visual observation. Palomo's method did not cause any complication of testicular atrophy and improved the fertility potential. PMID- 8427423 TI - Cadmium, lead, selenium, and zinc in semen of occupationally unexposed men. AB - Concentrations of cadmium, lead, selenium, and zinc were determined in semen and seminal plasma of 22 volunteers by atomic absorption spectrometry. Additionally conventional semen parameters and, by means of computer videomicrography, motion parameters of spermatozoa were evaluated. Concentrations of Cd, Pb, and Zn determined in semen were not significantly different from those measured in seminal plasma. However, selenium levels were significantly higher in semen (53.8 +/- 22.9 micrograms l-1) than in seminal plasma (40.4 +/- 15.5 micrograms l-1, P < 0.01). The investigated semen samples on average contained low levels of Cd (0.4 +/- 0.23 micrograms l-1) and Pb (9.8 +/- 6.5 micrograms l-1). Studies on the intra-individual variability revealed the following average coefficients of variation (%) for element concentrations: Pb (70), Cd (53), Se (27), and Zn (23); and for semen parameters: total sperm count (46), sperm concentration (37), motility (22), ejaculate volume (21), linearity (19), linear velocity (11), curvilinear velocity (10), and percentage of normally formed sperm (9). Significant positive correlations were detected between semen selenium levels and sperm concentration (r = 0.51, P < 0.05), and percentage of normally formed sperm (r = 0.46, P < 0.05), respectively. Sperm motility (r = 0.53, P < 0.02), linear (r = 0.76, P < 0.001) and curvilinear velocity (r = 0.64, P < 0.002) were significantly correlated with semen cadmium levels. PMID- 8427424 TI - Effect of blood pressure on hemorrhage volume and survival in a near-fatal hemorrhage model incorporating a vascular injury. AB - STUDY HYPOTHESIS: In a model of near-fatal hemorrhage that incorporates a vascular injury, stepwise increases in blood pressure associated with aggressive crystalloid resuscitation will result in increased hemorrhage volume and mortality. DESIGN: This study used a swine model of potentially lethal hemorrhage in the presence of a vascular lesion to compare the effects of resuscitation with mean arterial pressures of 40, 60, and 80 mm Hg. Twenty-seven fully instrumented immature swine (14.8 to 20 kg), each with a surgical-steel aortotomy wire in place, were bled continuously from a femoral artery catheter to a mean arterial pressure of 30 mm Hg. At that point the aortotomy wire was pulled, producing a 4 mm aortic tear and uncontrolled intraperitoneal hemorrhage. When the animal's pulse pressure reached 5 mm Hg, the femoral artery hemorrhage was discontinued and resuscitation was begun. INTERVENTIONS: Saline infusion was begun at 6 mL/kg/min and continued as needed to maintain the following desired endpoints: group 1 (nine) to a mean arterial pressure of 40 mm Hg, group 2 (nine) to a mean arterial pressure of 60 mm Hg, and group 3 (nine) to a mean arterial pressure of 80 mm Hg. After 30 minutes or a total saline infusion of 90 mL/kg, the resuscitation fluid was changed to shed blood infused at 2 mL/kg/min as needed to maintain the desired mean arterial pressure or to a maximum volume of 24 mL/kg. Animals were observed for 60 minutes or until death. MEASUREMENTS AND MAIN RESULTS: Data were compared using repeated-measures analysis of variance with a post hoc Tukey-Kramer, Fisher's exact test, and Kruskal-Wallis. Mortality was significantly greater in group 3 (78%) compared with either group 1 (11%; P = .008) or group 2 (22%; P = .028). Mean survival times were significantly shorter in group 3 (44 +/- 12 minutes) compared with either group 1 (58 +/- 6 minutes; P = .007) or group 2 (59 +/- 3 minutes; P = .006). The average intraperitoneal hemorrhage volumes were 13 +/- 14 mL/kg, 20 +/- 25 mL/kg, and 46 +/- 11 mL/kg for groups 1, 2, and 3, respectively (group 1 versus 2, P = .425; group 1 versus 3, P < .001; group 2 versus 3, P = .014). Group 2 animals demonstrated significantly greater oxygen deliveries compared with groups 1 and 3. CONCLUSION: In a model of near-fatal hemorrhage with a vascular injury, attempts to restore blood pressure with crystalloid result in increased hemorrhage volume and markedly higher mortality. PMID- 8427425 TI - Development of a piglet model of status epilepticus: preliminary results. AB - STUDY OBJECTIVE: Ventilation frequently is impaired during prolonged clinical seizures and their treatment. In a pilot study, respiratory, metabolic, and hemodynamic variables were studied during induced seizures in a lightly anesthetized, spontaneously breathing piglet model. PARTICIPANTS: Weanling, mixed breed domestic piglets. INTERVENTIONS: Piglets were instrumented with a tracheostomy, arterial catheter, and epidural electrodes. Conditions included hyperoxia, normothermia, and ketamine maintenance infusion throughout recordings. After baseline recordings, 2 mg/kg IV bicuculline was administered. For further model validation, piglets were randomized to infusions of diazepam (three), lorazepam (two), or saline (control; five) groups after ten minutes of untreated seizures. MEASUREMENTS AND MAIN RESULTS: Integrated tidal volume, respiratory rate, PaCO2, pH, arterial pressure, rectal temperature, heart rate, and bipolar EEG waveforms were recorded and compared at intervals for 60 minutes. Vigorous tonic-clonic seizures occurred in all piglets, confirmed by sudden synchronization and large-amplitude EEG waveforms. Increases in heart rate, arterial pressure, tidal volume, respiratory rate, PaCO2, minute ventilation, and base deficit occurred in all piglets during seizures as compared with baseline. Five minutes after bicuculline was administered, increases in minute ventilation (4.5 +/- 0.4 L/min at baseline to 13 +/- 2.1 L/min) were accounted for by increases in both tidal volume and respiratory rate. More abrupt decreases in respiratory rate were observed in anticonvulsant-treated piglets as compared with controls. The duration of continuous seizure activity (12 +/- 1.0 minutes versus 21 +/- 3.3 minutes; P < .05) was reduced in anticonvulsant-treated piglets. CONCLUSION: Significant increases in ventilation occur during generalized seizures in tracheostomized piglets given bicuculline. Diazepam and lorazepam infusions ameliorate seizure activity and suppress increases in respiratory rate but not minute ventilation as compared with controls. Problems with this model included baseline variability, temperature instability, and that direct respiratory stimulation from the convulsant agent may have occurred. PMID- 8427426 TI - Fructose-1,6-diphosphate fails to limit early myocardial infarction size in a canine model. AB - STUDY OBJECTIVE: Fructose-1,6-diphosphate (FDP) appears to improve early post myocardial infarction hemodynamics and limit early myocardial infarct size in previous canine studies. However, these studies did not account for the effect of collateral blood flow on infarct size. Our objective was to determine the effect of FDP on early infarct size and hemodynamics while measuring regional myocardial blood flow. DESIGN: A prospective, blinded, placebo-controlled laboratory study using a canine open-chest left anterior descending coronary artery (LAD) occlusion model. INTERVENTIONS: Twenty-two mongrel dogs were assigned randomly to receive either FDP (175 mg/kg, then 2 mg/kg/min for two hours) or placebo, beginning five minutes after LAD occlusion. MEASUREMENTS AND MAIN RESULTS: Regional myocardial blood flow, hemodynamics, and myocardial infarct size were determined. Infarct size was assessed using magnetic resonance imaging in a subset of animals. Three of the 22 dogs had no infarct and significantly higher collateral blood flow than the 19 animals with myocardial infarction (P < .001). Four hours after LAD occlusion, cardiac index, dP/dtmax, heart rate, and systolic and mean aortic pressures were not statistically different between groups. Infarct size expressed as area of necrosis/area at risk was similar between groups (FDP, 0.55 +/- 0.28; controls, 0.59 +/- 0.31). CONCLUSION: FDP given after occlusion of the LAD in this canine model did not limit early myocardial infarct size. PMID- 8427427 TI - Effective treatment of acute alkali injury of the rat esophagus with early saline dilution therapy. AB - BACKGROUND: Controversy persists regarding the appropriate treatment of acute alkali injury to the esophagus. The current study establishes a controlled model of alkali esophageal injury and examines the efficacy of saline dilution therapy. STUDY HYPOTHESIS: Early saline dilution therapy effectively reduces esophageal injury resulting from acute alkali exposure. METHODS: The esophagi were harvested from 60 Sprague-Dawley rats. Each end was cannulated with a 20-gauge catheter. Specimens were maintained in an oxygen-perfused saline bath (37 C) during a 60 minute experimental period and then fixed immediately in 10% formalin solution for histologic examination. Three experimental groups (A, B, and C) were perfused with 50% NaOH solution at time zero. Treatment with saline perfusion was performed immediately in group A, five minutes after injury in group B, and 30 minutes after injury in group C. The positive control group D was perfused with saline at time zero. A negative control, group E, was perfused with 50% NaOH at time zero. This group did not receive subsequent treatment with saline. Pathologic examination was performed in a blinded fashion using a score of 0 to 3 (0, no injury; 1, minimal; 2, moderate; 3, severe) for seven histologic criteria: epithelial viability, extent of injury, cornified epithelial cell differentiation, granular cell differentiation, epithelial cell nuclei, muscle cells, and muscle cell nuclei. RESULTS: The positive control group demonstrated scores of zero. Nonparametric analysis showed a significant difference among treatment groups for each injury category. Trend analysis revealed a significant progression of injury for each category associated with time to treatment. Discriminant analysis indicated that the muscle cells category was the most useful category with which to distinguish injury among groups. CONCLUSION: In our model, saline lavage decreased objective evidence of esophageal injury after a severe alkaline exposure, and early therapy enhanced this beneficial effect. PMID- 8427428 TI - Comparison of fluid infusion rates among peripheral intravenous and humerus, femur, malleolus, and tibial intraosseous sites in normovolemic and hypovolemic piglets. AB - STUDY OBJECTIVES: To compare infusion rates from various intraosseous sites (tibial, medial malleolar, distal femoral, and humeral) and at a peripheral IV site under gravity and pressure flow in normovolemic and hypovolemic states. DESIGN AND SETTING: A piglet model was used to assess rates of infusion under varying conditions in a university hospital animal laboratory. Analysis of variance was used to evaluate site differences. PARTICIPANTS: Twenty-three Yorkshire-Landrace mix pigs (weight, 12 to 23 kg) were studied. INTERVENTIONS: Animals were anesthetized and intubated before cannulation with 18-gauge bone marrow needles at intraosseous sites and 22-gauge Teflon catheters in peripheral vessels. Infusion rates under gravity and 300 mm Hg pressure were determined. Infusion rates under similar conditions were repeated in hypovolemic animals with acute bleeding of 25 mL/kg. MEASUREMENTS AND MAIN RESULTS: Mean infusion rates (mL/min) for gravity versus 300 mm Hg pressure in normovolemic pigs were 13.1 versus 40.9 for peripheral IV, 11.1 versus 41.3 for humerus, 9.3 versus 29.5 for femur, 8.2 versus 24.1 for malleolus, and 4.3 versus 17.0 for tibia. Hypovolemia resulted in average decreased rates of 32%. Infusion rates were significantly different between sites and between normovolemia and hypovolemia (P = .0001). CONCLUSION: Intravenous access is the most efficacious method of acute volume replacement. Intraosseous sites differ in the infusion rates obtained--descending order is humerus, femur, malleolus, and tibia, but each is a reasonable alternative for short-term vascular access. PMID- 8427429 TI - Effectiveness and safety of intravenous nalmefene for emergency department patients with suspected narcotic overdose: a pilot study. AB - STUDY OBJECTIVE: To evaluate the efficacy and safety of nalmefene, an investigational narcotic antagonist that has potential advantages over naloxone because of its four- to eight-hour half-life, in emergency department patients with possible narcotic overdose. DESIGN: Multi-institutional, prospective, phase II, open-label study. TYPE OF PARTICIPANTS: Complete data were available for 53 cases from two teaching hospitals. Men 18 years old or older who would otherwise receive naloxone were eligible (two women were enrolled inadvertently). METHODS: Over four hours, one to ten boluses (median, one) of 0.5 or 1.0 mg nalmefene IV were given as often as every two minutes based on clinical need. Respirations, blood pressure, pulse, pupil size, and overall clinical response were monitored. Overall clinical response (1, no change; 2, partial response; 3, complete response), first assessed at two minutes, was analyzed by the Mann-Whitney U test. RESULTS: Fifteen of 25 (0.5 mg) and nine of 28 (1.0 mg) cases were opiate positive. Twelve of 15 (0.5 mg) and six of nine (1.0 mg) opiate-positive cases had a rapid complete response. Coincident causes of depressed sensorium were identified in the remaining six opiate-positive cases. No difference in initial overall clinical response was seen between 0.5-mg and 1.0-mg opiate-positive cases (P = .59). No deterioration requiring repeat nalmefene occurred in opiate positive cases, even if methadone (four), codeine (two), or pentazocine (one) was found. No serious adverse events were judged to be related to nalmefene. CONCLUSION: Nalmefene is effective in the reversal of opiate overdose and appears to be safe in the management of patients with altered sensorium. PMID- 8427430 TI - The efficacy of metoclopramide in the treatment of migraine headache. AB - STUDY OBJECTIVES: By evaluating the efficacy of metoclopramide alone and in combination with ibuprofen versus placebos, this study was designed to both evaluate the efficacy of metoclopramide and elucidate its mechanism of action in the treatment of migraine headache. DESIGN: The study was conducted over a two year period and was a randomized, double-blind, placebo-controlled study. SETTING: An urban teaching hospital. PARTICIPANTS: Patients enrolled were at least 18 years old and had recurring headaches with one or more of the following characteristics: unilateral, preceded by neurologic symptoms, significant nausea and vomiting, or mood changes and photophobia. INTERVENTION: Ten milligrams of metoclopramide or an equal volume of IV normal saline was given and 600 mg of ibuprofen or identical-appearing placebo was given orally at time 0. Patients rated their pain and nausea at time 0, 30 minutes, and 60 minutes using visual analog scales. RESULTS: The differences in pain and nausea scores for the metoclopramide + placebos group versus the other three groups were tested using exact nonparametric (Mann-Whitney) statistical procedures. The metoclopramide + placebos group had significantly better relief of pain compared with the placebos + ibuprofen and placebos + placebos groups. The metoclopramide + placebos group had significantly better relief of nausea than the ibuprofen + placebos group; nausea scores for the placebos + placebos group could not be analyzed due to excessive variance from the other groups at baseline. The differences between the metoclopramide + placebos group and the metoclopramide + ibuprofen group were not statistically significant with regard to either pain or nausea. CONCLUSION: Metoclopramide is efficacious in the treatment of both the pain and nausea of migraine headache. This is a direct action that is not dependent on the concomitant administration of another agent. PMID- 8427431 TI - A one-year evaluation of calcium channel blocker overdoses: toxicity and treatment. AB - STUDY OBJECTIVE: To examine the cardiovascular toxicity of calcium channel blockers and the efficacy of various treatments. DESIGN: Case series collected prospectively over one year. SETTING: Three regional poison control centers. TYPE OF PARTICIPANTS: One hundred thirty-nine hospitalized patients who had ingested a calcium channel blocker. INTERVENTIONS: Calcium, dopamine, atropine, isoproterenol, glucagon, and pacemakers. MAIN RESULTS: Hypotension, sinus node suppression, and dysrhythmias often occur with calcium channel blocker overdoses, but atrioventricular nodal block occurs more often with verapamil (chi 2 test, P < .025). Calcium was administered to 23 patients and was efficacious in reversing depression of cardiac conduction and increasing blood pressure. Dopamine was administered to ten patients and was efficacious in increasing blood pressure. Atropine was administered to eight patients, but only two had a positive response. CONCLUSION: Atrioventricular nodal depression is more common with verapamil overdoses. Calcium and dopamine are useful in treating toxicity from calcium channel blocker overdose, whereas atropine is sometimes useful. PMID- 8427432 TI - Comparison of midazolam and diazepam for conscious sedation in the emergency department. AB - STUDY OBJECTIVE: To compare the efficacy of diazepam and midazolam when used for conscious sedation in emergency department patients. DESIGN: Prospective, randomized, double-blind, multicenter trial. SETTING: Three university EDs. TYPE OF PARTICIPANTS: Patients requiring one of the following procedures: abscess drainage, joint reduction, extensive suturing, chest tube insertion, or lumbar puncture. INTERVENTIONS: Diazepam (2.5 mg/mL) or midazolam (1 mg/mL) was administered until the desired level of sedation was achieved to a maximum of 5 mL. Fentanyl citrate was administered if needed for pain. MEASUREMENTS AND MAIN RESULTS: Thirty-three patients received diazepam and 36 received midazolam. Patients receiving midazolam had a greater degree of early sedation (P < .05), a higher 90-minute alertness scale score (P < .05), more patients ready for discharge at 90 minutes (P = .05), significantly less recall for the procedure (P < .02), and less pain on injection (P < .01) than patients who were given diazepam. CONCLUSIONS: Diazepam and midazolam are both effective for conscious sedation in ED patients. Midazolam causes less pain on injection, a significantly greater degree of early sedation, and a more rapid return to baseline function. PMID- 8427433 TI - Comparison of intramuscular meperidine and promethazine with and without chlorpromazine: a randomized, prospective, double-blind trial. AB - STUDY OBJECTIVES: To compare the effectiveness of intramuscular meperidine (2 mg/kg) and promethazine (1 mg/kg) with chlorpromazine (MPC) or without chlorpromazine (MP) (1 mg/kg) for sedation of children undergoing emergency department procedures. DESIGN: Randomized, double-blind trial. SETTING: A community and university hospital ED. TYPE OF PARTICIPANTS: Eighty-seven hemodynamically and neurologically stable children less than 16 years old. INTERVENTIONS: IM sedation followed by intended procedure. MEASUREMENTS AND MAIN RESULTS: Children receiving either combination were not significantly different with regard to age, sex, weight, chronic illness, and indications. Procedures included laceration repair (46), fracture reduction (25), and others (16). Mean onset of action was similar (16 +/- 12 minutes), whereas the duration of action was significantly longer after MPC (63 +/- 57 minutes [mean +/- SD] compared with MP 29 +/- 36 minutes; P < .05, Student's t-test). Paradoxical hyperactivity occurred only after MP (three of 43 cases; P = NS, Fisher's exact test), whereas transient oxygen desaturation occurred only after MPC (one of 44 cases; P = NS). No other serious complications were observed. Three observers rated the effectiveness of sedation and analgesia on separate 10.2-cm visual-analog scales. Overall, MPC received significantly better ratings (7.4 +/- 2.1 cm) than MP (5.7 +/- 3.0 cm; P < .05, Mann-Whitney U test). Parents believed sedation worked well in 90% of cases. Their children had bad memories of the procedure in only 9% of cases. CONCLUSION: Elimination of chlorpromazine from the IM combination of meperidine and promethazine for pediatric sedation during ED procedures results in a significant reduction in efficacy. PMID- 8427434 TI - Dexamethasone as adjuvant therapy for severe acute pharyngitis. AB - STUDY OBJECTIVE: To determine the efficacy of dexamethasone as adjuvant therapy to improve pain relief in patients with severe, acute exudative pharyngitis. DESIGN: Prospective, randomized, double-blinded, placebo-controlled clinical trial. SETTING: Large, urban community hospital emergency department with an emergency medicine residency program. TYPE OF PARTICIPANTS: Patients aged 12 to 65 years old with exudative pharyngitis and severe dysphagia/odynophagia. Patients with cancer, AIDS, diabetes mellitus, recent steroid use, pregnancy, or suspicion of peritonsillar abscess were excluded. INTERVENTIONS: All patients received oral penicillin (500 mg Pen VK) or erythromycin (333 mg base) three times daily for ten days in addition to either 10 mg single-dose dexamethasone or saline placebo IM injection. MEASUREMENTS AND RESULTS: Fifty-eight patients graded their initial degree of throat pain on a visual-analog scale that was 15 cm long and scored from 0 to 3.0 in 0.5-cm increments. Follow-up was obtained on 51 patients to determine their condition at 24 hours. At entry, there was no difference in age, weight, antibiotic assignment, or initial pain score between groups. Improvement in pain score (initial versus 24 hours) was 1.8 +/- 0.8 in the 26 patients of the dexamethasone group and 1.2 +/- 0.9 in the 25 patients of the placebo group (P < .05). Time to onset of pain relief was also faster in steroid-treated patients who demonstrated relief beginning at 6.3 +/- 5.3 hours, compared with 12.4 +/- 8.5 hours in the placebo group (P < .01). Of the 26 patients evaluated at seven days (13 in each group), time to complete lack of pain averaged 15.0 +/- 11.4 hours in the dexamethasone group and 35.4 +/- 17.9 hours in the placebo group (P < .02). Complications attributable to dexamethasone were not observed. CONCLUSION: In patients with severe, acute exudative pharyngitis, single-injection dexamethasone adjuvant compared with placebo resulted in statistically and clinically significant improvement, as evidenced by more rapid onset and greater degree of pain relief. PMID- 8427435 TI - Nonvenomous snakebite in Massachusetts: prophylactic antibiotics are unnecessary. AB - STUDY OBJECTIVE: To investigate wound infection in nonvenomous snakebite. DESIGN: Prospective clinical series. SETTING: Massachusetts Poison Control System. PARTICIPANTS: Seventy-two consecutive children and adults with nonenvenomated snakebite wounds. INTERVENTIONS: Patients were examined and advised that there was no published clinical evidence that prophylactic antibiotics were beneficial for nonenvenomated snakebites. They were re-examined five to ten days after the bite. MAIN RESULTS: Four patients used prophylactic antibiotics; 68 did not. There were no wound infections. Plain radiographs revealed a snake tooth fragment in the persistently tender wound of one patient bitten by his pet python. CONCLUSION: Despite reports that pathogenic bacteria can be isolated from snakes' mouths, nonenvenomated snakebites in Massachusetts usually do not require prophylactic antibiotic therapy. Radiography is occasionally indicated for persistently tender wounds inflicted by large snakes. PMID- 8427436 TI - Are victims of injury sometimes victimized by attempts at fluid resuscitation? PMID- 8427437 TI - Intentional massive insulin overdose: recognition and management. AB - A case of intentional massive insulin overdose requiring prolonged glycemic support is presented. Suicidal insulin overdose may be more common than generally appreciated. Because hypoglycemic reactions are evaluated routinely in the ED, emergency physicians should maintain a high degree of suspicion regarding suicidal intent or foul play in diabetics with hypoglycemia who respond minimally to the administration of concentrated glucose solutions or in hypoglycemic presentations by nondiabetics who have access to diabetic medications. Fingerstick glucose evaluations or serum glucose levels should be obtained routinely at 15 to 30 minutes after glucose administration in any hypoglycemic patient to gauge the intensity of glucose use. Inability to maintain euglycemia following glucose administration suggests excessive insulin and requires further workup. Evaluation of serum insulin and C-peptide levels is useful in confirming intentional overdoses in cases that are not clear-cut. Glucose infusion rates must be tailored individually to each overdose situation as great individual variability exists in insulin absorption and effects. The clinician should anticipate the possible need for prolonged glycemic support in this setting. PMID- 8427438 TI - An experimental algorithm versus standard advanced cardiac life support in a swine model of out-of-hospital cardiac arrest. AB - STUDY OBJECTIVE: To compare an experimental algorithm with standard advanced cardiac life support in a swine model of out-of-hospital cardiac arrest. DESIGN: Randomized, controlled experimental trial. SETTING/TYPE OF PARTICIPANT: Animal laboratory using swine. INTERVENTIONS: Eighteen swine (17.8 to 23.7 kg) were sedated, intubated, anesthetized, and instrumented for monitoring of arterial and central venous pressures and ECG. Ventricular fibrillation was induced using a bipolar pacing catheter. Animals were randomized to treatment with the experimental algorithm or standard advanced cardiac life support therapy after eight minutes of untreated ventricular fibrillation. The experimental algorithm consisted of starting CPR; giving high-dose epinephrine (0.20 mg/kg), lidocaine (1.0 mg/kg), bretylium (5.0 mg/kg), and propranolol (0.5 to 1.0 mg) by peripheral IV; hyperventilating (20 to 25 breaths per minute); and delaying countershock (5 J/kg) 60 seconds after completion of drug delivery. Data were analyzed with the Student's t-test and Fisher's exact test. MEASUREMENTS AND MAIN RESULTS: Outcome variables were arterial and central venous pressures, return of spontaneous circulation, and one-hour survival. Hemodynamics were not different between groups during CPR. Return of spontaneous circulation occurred in seven of nine swine (77%) in the experimental algorithm group versus two of nine swine (22%) in the advanced cardiac life support group (P = .057). Four of nine swine (44%) in the experimental algorithm group survived to one hour versus none of the animals in the advanced cardiac life support group (P = .041). CONCLUSION: In this swine model of out-of-hospital cardiac arrest, animals treated with an experimental algorithm had a significant improvement in one-hour survival compared with those treated with advanced cardiac life support. PMID- 8427439 TI - Two-thumb versus two-finger chest compression during CRP in a swine infant model of cardiac arrest. AB - STUDY OBJECTIVE: To test the hypothesis that two-thumb chest compression generates higher arterial and coronary perfusion pressures than the current American Heart Association-approved two-finger method. DESIGN: Randomized, crossover experimental trial. SETTING AND PARTICIPANTS: Animal laboratory experiment with seven swine of either sex weighing 9.4 kg (SD, 0.8 kg), representing infants less than 1 year old. INTERVENTIONS: Animals were sedated with IM ketamine/xylazine, intubated with a 6.0 Hi-Lo endotracheal tube, anesthetized with alpha-chloralose, and paralyzed with pancuronium. ECG was monitored continuously. Left femoral arterial and Swan-Ganz catheters were placed. Cardiac arrest was induced with an IV bolus of KCl and verified by ECG and pressure tracings. Five American Heart Association-certified basic rescuers were randomly assigned to perform external chest compressions for one minute by either the currently recommended two-finger method or the two-thumb and thorax squeeze method. After all five completed their first trial, rescuers crossed over to the other method for a second minute of compressions. Ventilation was performed with a bag-valve device, and no drugs were given during CPR. After three complete cycles, the fourth through sixth cycles of compressions were recorded. Every compression was analyzed for arterial systolic, diastolic, mean, and coronary perfusion pressures. One thousand fifty compressions were analyzed with repeated-measures analysis of variance and Scheffe multiple comparisons. RESULTS: Systolic blood pressure, diastolic blood pressure, mean arterial pressure, and coronary perfusion pressure were all significantly higher (P < .001) with the two-thumb thoracic squeeze technique: systolic blood pressure, 59.4 versus 41.6 mm Hg; diastolic blood pressure, 21.8 versus 18.5 mm Hg; mean arterial pressure, 34.2 versus 26.1 mm Hg; and coronary perfusion pressure, 15.1 versus 12.2 mm Hg. CONCLUSION: The two-thumb method of chest compression generates significantly higher arterial and coronary perfusion pressures than the two-finger method in this infant model of cardiac arrest. PMID- 8427440 TI - Angiotensin II effects in a swine model of cardiac arrest. AB - STUDY OBJECTIVE: To evaluate the effect of different doses of the vasopressor angiotensin II on aortic diastolic and coronary perfusion pressures in a swine model of cardiac arrest. DESIGN: Immature swine were anesthetized and hemodynamically monitored. Ventricular fibrillation was induced; CPR was begun after ten minutes of ventricular fibrillation. Aortic and right atrial pressures were recorded continuously throughout the experiment. SETTING: Animal research laboratory. PARTICIPANTS: Immature swine weighing 21 to 32 kg. INTERVENTIONS: Angiotensin II was administered in doses of 25, 50, 75, and 100 micrograms/kg at 13 minutes of ventricular fibrillation. Each dose was tested in four swine. RESULTS: Except for animals receiving 25 micrograms/kg, aortic diastolic pressure increased significantly in all groups after administration of angiotensin II compared with groups receiving CPR alone. Coronary perfusion pressure increased significantly only with 50- and 75-micrograms/kg doses. There were no significant differences in hemodynamic parameters among the different dose groups. CONCLUSION: Angiotensin II in doses of 50, 75, and 100 micrograms/kg significantly increased aortic diastolic pressure, and doses of 50 and 75 micrograms/kg significantly increased coronary perfusion pressure in this model of cardiac arrest. PMID- 8427441 TI - Diffuse subcutaneous emphysema, pneumomediastinum, and pneumothorax after dental extraction. AB - Subcutaneous emphysema, pneumomediastinum, and pneumothorax may result from surgical procedures and trauma and usually do not present a diagnostic dilemma. We present a case of subcutaneous emphysema, pneumomediastinum, pneumothorax, and pneumoretroperitoneum after a dental procedure with an air-and-water-cooled turbine burr drill. This allowed air and water under pressure to be driven into the field and track through the fascial planes. Although this is a common occurrence, these patients frequently go undiagnosed or misdiagnosed as allergic reactions to locally administered anesthetic agents. If a large amount of air is injected, it may track into not only the subcutaneous tissues but also the mediastinum, pleural space, and retroperitoneal space. Patients with significant amounts of air must be admitted, observed for airway compromise, and be provided IV antibiotics and hydration. PMID- 8427442 TI - Evaluation and management of acute uvular edema. AB - Acute uvular edema is a potentially life-threatening condition with a variety of causes. Other upper airway structures, most notably the epiglottis, may also become involved, and complete or partial airway obstruction may ensue rapidly. An accurate diagnosis may be essential in establishing effective treatment. PMID- 8427443 TI - Lemierre syndrome: the forgotten disease. AB - Lemierre syndrome is characterized by an acute oropharyngeal infection, suppurative thrombophlebitis of the internal jugular vein, anaerobic sepsis, and metastatic infections. Before the discovery of antibiotics, this disease usually was fatal. We report the case of a patient with all of these findings, indicating that Lemierre syndrome still is seen today. Prompt recognition, abscess drainage, and appropriate antibiotic coverage result in complete recovery in most patients. PMID- 8427444 TI - Tension pneumocephalus related to an epidermoid tumor of ethmoid sinus origin. AB - A 37-year-old man presented with a four-week history of progressive left-sided weakness, frontal headache, confusion, and drowsiness. A computed tomography evaluation of the head revealed a 7-cm pneumatocele in the right frontoparietal region with shift of the falx to the left. The patient was taken to surgery, where the pneumatocele was decompressed and an epidermoid tumor that originated in the left ethmoid sinus was removed. After the operation, the presenting symptoms resolved promptly. PMID- 8427445 TI - Stroke following rottweiler attack. AB - A previously healthy 50-year-old man suffered a major right cerebral infarction shortly after receiving head and neck bites in an attack by two rottweilers. Arteriography revealed occlusion of the right middle cerebral artery, an intimal flap and pseudoaneurysm in the high right internal carotid artery just proximal to the skull base, and an obstructed right vertebral artery. The powerful jaw mechanism of rottweilers and other large mastiff-type dogs makes their bites particularly destructive. The predilection of these animals for attacking the head and upper body makes occult crush injury to the extracranial cerebral vessels an important diagnostic consideration following such attacks. PMID- 8427446 TI - Animal use in research. American College of Emergency Physicians. PMID- 8427447 TI - Telephone orders in the emergency department. American College of Emergency Physicians. PMID- 8427448 TI - The use of facsimile machines and electronic data transfer in the emergency department. American College of Emergency Physicians. PMID- 8427449 TI - Timeliness of emergency imaging. American College of Emergency Physicians. PMID- 8427450 TI - Plasma beta-endorphin levels in acute myocardial infarction. PMID- 8427451 TI - Concordance versus correlation. PMID- 8427452 TI - Use of infected cultured cells to compare ultrastructural features of Neospora caninum from dogs and Toxoplasma gondii. AB - The ultrastructure of tachyzoites of 3 isolates of Neospora caninum from dogs was examined, using transmission electron microscopy of infected cultured cells. Ultrastructure of the 3 isolates was similar. Tachyzoites had a pellicle, 22 subpellicular microtubules, a conoid, anterior and posterior polar rings, 8 to 12 electron-dense rhoptries, numerous micronemes, a single vesicular nucleus, tubular mitochondria, Golgi complexes, ribosomes, endoplasmic reticula, an inactive micropore, electron-dense bodies, lipid bodies, and amylopectin bodies. Most tachyzoites were located adjacent to the host cell nucleus in a parasitophorous vacuole that contained numerous intravacuolar tubules. Tachyzoites divided by endodyogeny. Neospora caninum tissue cysts were not seen. Comparison of N caninum with Toxoplasma gondii tachyzoites indicated that the 2 species can be differentiated on the basis of structure and numbers fo rhoptries and numbers and location of micronemes and electron-dense bodies. PMID- 8427453 TI - Development of Anaplasma marginale in salivary glands of male Dermacentor andersoni. AB - Development of the rickettsia, Anaplasma marginale, in salivary glands of male Dermacentor andersoni exposed as nymphs or adult ticks, was studied indirectly by inoculation of susceptible calves with homogenates and directly by examination, using light microscopy and a DNA probe; some unfed ticks were incubated before tissues were collected. Salivary gland homogenates made from ticks in every treatment group caused anaplasmosis when injected into susceptible calves; prepatent periods decreased as the time that ticks had fed increased. Colonies of A marginale were seen only in salivary glands of ticks exposed as adults and not in those exposed as nymphs; the percentage of salivary gland acini infected in these ticks increased linearly with feeding time. However, the probe detected A marginale DNA in salivary glands of ticks from both groups; the amount of DNA detected increased as feeding time was extended. The amount of A marginale DNA appeared to remain constant in gut tissues, but to increase in salivary glands. Salivary glands of adult-infected male ticks that were incubated, but did not feed a second time, became infected with A marginale, and the pattern of infection of acini varied with incubation temperature. Development of A marginale in salivary glands appears to be coordinated with the tick feeding cycle; highest infection rate was observed in ticks exposed as adults. PMID- 8427454 TI - Determination of virulence and pathogenesis of a canine strain of Leishmania leishmania infantum in hamsters and dogs. AB - Visceral leishmaniasis was experimentally induced in hamsters by the intracardiac inoculation of 10(7) amastigotes of Leishmania leishmania infantum of canine origin. At postinoculation (PI) days 7, 21, 42, and 63, hamsters were euthanatized. Body weights and total parasite numbers of the liver and spleen were determined. Gross and histologic evaluations of tissues were done. Dogs also were inoculated IV with 10(8) amastigotes/kg of body weight. Samples were obtained from dogs prior to infection and at biweekly PI intervals for CBC and serum chemical analysis, for lymphocyte blastogenic assay by use of blood leukocytes, and for ELISA to determine antileishmanial antibody titers. At PI week 12, dogs were necropsied; organ weights, tissue imprints of the liver and spleen, and histologic interpretations of tissues were obtained. Hamsters developed high parasite numbers within 7 days after inoculation, at which time the total parasite numbers in the liver (3.51 x 10(7) amastigotes) was observed to be approximately 11 times that in the spleen (2.93 x 10(6)). The liver had the highest parasite numbers throughout the infection period. Some infected hamsters became either cachectic and emaciated or ascitic. Two of the 10 infected hamsters died at PI days 54 and 58. Moderate to severe hepatosplenomegaly with granulomatous inflammatory reactions characterized by the presence of varied numbers of parasitized macrophages, giant cells, and hepatic Schaumann bodies were observed in infected hamsters. Infected dogs developed significantly altered hematologic values consisting of mild anemia and moderate leukopenia at PI weeks 8 to 12. Hyperproteinemia characterized by hyperglobulinemia (4.5 g/dl) was noticed at PI week 4.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427455 TI - Effects of orally administered prednisone on glucose tolerance and insulin secretion in clinically normal dogs. AB - Prednisone was administered orally for 4 weeks at a dosage of 1.1 mg/kg of body weight/d, in divided dose every 12 hours, to a group of healthy adult dogs (n = 12). Intravenous glucose tolerance testing was performed before and after the 28 day regimen in each dog, as well as in dogs of a control group (n = 6). Glucose metabolism was evaluated by measurement of preprandial plasma insulin and glucose concentrations, total insulin secretion, and fractional clearance of glucose. Mean preprandial plasma insulin and glucose concentrations were not increased after the 4-week regimen of prednisone. Total insulin secretion in response to an IV administered glucose load was not increased in treated dogs, compared with pretreatment values or with values for control dogs. The fractional clearance of glucose was also not altered in dogs given prednisone. Results indicate that anti inflammatory doses of prednisone, given orally for 4 weeks, probably do not alter insulin sensitivity or glucose tolerance in clinically normal dogs. PMID- 8427456 TI - Serodiagnosis of paratuberculosis in sheep by use of agar gel immunodiffusion. AB - An agar gel immunodiffusion (AGID) test was used over a 3-year period to examine 1,871 serum samples from sheep representing 5 Mycobacterium paratuberculosis infected flocks and 4 flocks presumed to be uninfected. Of 1,032 sheep, 31 had positive AGID test results (scoring 1 to 5), and 23 of these 31 were necropsied. Infection with M paratuberculosis was confirmed by 1 or more of the following findings: observation of typical lesions on histologic examination of sections of ileum or ileocecal lymph nodes, observation of clumps of acid-fast bacteria in mucosal smears of ileum, and isolation of the organism from feces or tissue. False-positive results on AGID testing were not found in sheep from flocks known to have exposure to Corynebacterium pseudotuberculosis. Diarrhea in infected sheep was observed infrequently; chronic, severe weight loss was the most common sign observed. On histologic examination of tissues from 20 infected sheep, 16 (80%) had diffuse lesions of the ileum and 13 (65%) had acid-fast bacteria in areas of ileal inflammation; 4 had discrete granulomas and peripheral lymphocytic infiltrates in the ileum. Sheep with diffuse lesions tended to have higher mean scores on AGID testing and examination for acid-fast bacteria, compared with those from sheep with more discrete lesions. Bacteriologic culture yielded M paratuberculosis from only 3 sheep with paratuberculosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427457 TI - Effects of oral administration of anti-inflammatory doses of prednisone on thyroid hormone response to thyrotropin-releasing hormone and thyrotropin in clinically normal dogs. AB - Prednisone was given orally to 12 dogs daily for 35 days at an anti-inflammatory dosage (1.1 mg/kg of body weight in divided dose, q 12 h) to study its effect on thyroxine (T4) and triiodothyronine (T3) metabolism. Six of these dogs were surgically thyroidectomized (THX-Pred) and maintained in euthyroid status by daily SC injections of T4 to study peripheral metabolism while receiving prednisone; 6 dogs with intact thyroid gland (Pred) were given prednisone; and 6 additional dogs were given gelatin capsule vehicle as a control group (Ctrl). Baseline T4 concentration after 4 weeks of treatment was not significantly different in dogs of the THX-Pred or Pred group (mean +/- SEM, 2.58 +/- 0.28 or 3.38 +/- 0.58 micrograms/dl, respectively) vs dogs of the Ctrl group (2.12 +/- 0.30 micrograms/dl). A supranormal response of T4 to thyrotropin was observed in dogs of the Pred group, but the T4 response to thyrotropin-releasing hormone was normal. Baseline T3 concentration in dogs of both steroid-treated groups was significantly (P < 0.05) lower after 2 and 4 weeks of prednisone administration vs pretreatment values, but normalized 2 weeks after prednisone was stopped. Free T3 (FT3) and T4 (FT4) fractions and absolute FT3 and FT4 concentrations were not altered by prednisone administration. Reverse T3 (rT3) concentration in vehicle treated Ctrl dogs (26.6 +/- 3.5 ng/dl) was not different from rT3 concentration in dogs of the THX-Pred (25.7 +/- 4.3 ng/dl) and Pred (28.9 +/- 3.8 ng/dl) groups after 4 weeks of medication.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427458 TI - Pulmonary artery wedge pressure increases with high-intensity exercise in horses. AB - Using catheter mounted microtip manometers, right atrial, pulmonary artery, and pulmonary artery wedge pressures were studied in 8 horses while they were standing quietly (rest), and during galloping at treadmill speeds of 8, 10, and 13 m/s. At rest, mean (+/- SEM) heart rate, mean right atrial pressure, mean pulmonary artery pressure, and mean pulmonary artery wedge pressure were 37 (+/- 2) beats/min, 8 (+/- 2) mm of Hg, 31 (+/- 2) mm of Hg, and 18 (+/- 2) mm of Hg, respectively. Exercise at treadmill belt speed of 8 m/s resulted in significant (P < 0.05) increments in heart rate, right atrial pressure, pulmonary artery systolic, mean, diastolic and pulse pressures, and pulmonary artery wedge pressure. All these variables registered further significant (P < 0.05) increments as work intensity increased to 10 m/s, and then to 13 m/s. Pulmonary artery diastolic pressure was, however, not different among the 3 work intensities. During exercise at belt speed of 13 m/s, heart rate, mean right atrial pressure, mean pulmonary artery pressure, pulmonary artery pulse pressure, and mean pulmonary artery wedge pressure were 213 (+/- 5) beats/min, 44 (+/- 4) mm of Hg, 89 (+/- 5) mm of Hg, 69 (+/- 4) mm of Hg, and 56 (+/- 4) mm of Hg, respectively. Assuming mean intravascular pulmonary capillary pressure to be halfway between the mean pulmonary arterial and venous pressures, its value during exercise at 13 m/s may have approached 72.5 mm of Hg. Transmural pressure (intravascular minus alveolar pressure) across pulmonary capillaries may be even higher because of the large negative pleural pressure swings in galloping horses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427459 TI - Kinetic analysis of D-xylose distribution after intravenous administration to mares. AB - Multicompartmental analysis was applied to study the kinetics of D-xylose distribution after IV administration to healthy mares deprived of food for 12 and 96 hours. Urinary excretion of D-xylose was measured over a 15-hour period after administration. The plasma D-xylose concentrations in this study were in the range found after oral tolerance testing. The disposition of D-xylose was described by a two-compartment model with linear kinetic characteristics. Total volume of distribution decreased significantly (P < 0.025) from 0.270 L/kg of body weight after the 12-hour period of food deprivation to 0.235 L/kg after the 96-hour period. Fractional rate of transfer between the central and peripheral compartments did not change after 96 hours without food. Approximately a third of the D-xylose administered was recovered in the urine. Difference in urinary elimination between the 12- and 96-hour periods was not significant. Nonrenal elimination rate was determined to be twice the renal elimination rate. The results indicated that formal kinetic analysis can provide useful information about D-xylose distribution in horses. The decreased D-xylose space found after a 96-hour period of food deprivation would tend to increase the plasma D-xylose concentration, and this may help in the interpretation of the D-xylose absorption test applied to anorectic horses. PMID- 8427460 TI - Characteristics of L-glutamine transport in equine jejunal brush border membrane vesicles. AB - The sodium-dependent transporter system responsible for L-glutamine uptake by brush border membrane vesicles prepared from equine jejunum was characterized. Vesicle purity was ascertained by a 14- to 17-fold increase in activity of the brush border enzyme markers. Glutamine uptake was found to occur into an osmotically active space with negligible membrane binding. The sodium-dependent velocity represented approximately 80% of total uptake and demonstrated overshoots. Kinetic studies of sodium-dependent glutamine transport at concentrations between 5 microM and 5 mM revealed a single saturable high affinity carrier with a Michaelis constant of 519 +/- 90 microM and a maximal transport velocity of 3.08 +/- 0.97 nmol/mg of protein/10 s. Glutamine uptake was not affected by changes in environmental pH. Lithium could not substitute for sodium as a contransporter ion. 2-Methylaminoisobutyric acid inhibited the sodium dependent carrier only minimally, but marked inhibition (> 90%) was observed in the presence of histidine, alanine, cysteine, and nonradioactive glutamine. Kinetic analysis of the sodium-independent transporter revealed it to have a Michaelis constant = 260 +/- 47 microM and a maximal transport velocity of 0.32 +/- 0.06 nmol/mg of protein/10 s. We conclude that glutamine transport in equine jejunal brush border membrane vesicles occurs primarily via the system B transporter and, to a lesser extent, by a sodium-independent carrier. PMID- 8427461 TI - Fetal heart rate patterns and the influence of myometrial activity during the last month of gestation in cows. AB - Five cows in the last month of gestation, provided with uterine electrodes and in which catheters had been chronically installed in the fetal aorta, were used to study patterns of fetal heart rate (FHR) and the influence of periods of myometrial electrical activity during gestation (contractures) on FHR. The FHR was calculated by counting the number of blood pressure pulses on the tracings during alternate periods of 12 seconds. Three 1-hour recordings without contractures and 10 recordings during the time of a contracture were randomly selected for each cow. The calculated data points were plotted on a graph to display FHR patterns. In 41 periods associated with single contractures, FHR data points were taken every 72 seconds. Changes in absolute and relative FHR in these periods were determined to analyze a possible effect of contractures on FHR. Three types of variation in FHR patterns could be distinguished: a short-term, low-amplitude variation of basal FHR; a second type in which the duration was < 4 minutes and the amplitude was > or = 15 beats/min; and prolonged periods with increased or decreased FHR values (> 4 minutes and > or = 15 beats/min). The relationship between these types of variation and fetal activity states remains to be established for cows. During the 60 hours of recordings that were analyzed, a period of several minutes during which FHR values were extremely high (> 180 beats/min) was found 3 times. There were no significant differences in absolute or relative FHR before, during, or after a contracture. PMID- 8427462 TI - Pulmonary response to intravenous administration of 5-hydroxytryptamine after type-2 receptor blockade in healthy calves. AB - The purpose of this study was to investigate the mechanism by which 5 hydroxytryptamine (5-HT) modifies respiratory function, specifically, hyperventilation, diffuse bronchoconstriction, and pulmonary arterial hypertension in cattle. We determined whether the IV response to 5-HT in calves was attributable to stimulation of 5-HT2 receptors. Six healthy unsedated young bull calves of the Friesian (n = 4) and of the Belgian White and Blue (n = 2) breeds were used. A specific 5-HT2 antagonist (metrenperone, 0.05 mg/kg of body weight) was administered IM 30 minutes before the cattle were given a 5-minute IV 5-HT infusion. Pulmonary function values were registered before, during, and after the 5-HT challenge infusion. Minute volume increased significantly, because of an increase in respiratory rate. Conversely, lung dynamic compliance, total pulmonary resistance, and pulmonary arterial pressure were not changed. We concluded that in cattle, 5-HT-induced ventilatory response is not mediated through activation of 5-HT2 receptors. However, the 5-HT2 receptors are involved in 5-HT-induced broncho- and pulmonary vasoconstriction. PMID- 8427463 TI - Refractive state of aphakic and pseudophakic eyes of dogs. AB - Streak retinoscopy was performed by 5 ophthalmologists on 256 eyes (191 dogs) to determine their postoperative refractive state after cataract extraction. Aphakic and pseudophakic eyes that had been implanted with 1 of 5 intraocular lenses (IOL) with dioptric powers ranging from +14.5 to +38 diopters (D) were studied. By use of ANOVA, breed and body type of dog and individual performing refraction were found to have no detectable effect on final refractive state. Mean refractive state of aphakic eyes was +14.4 +/- 2.10 D. Mean refractive state for different IOL powers was as follows: +14.5 D IOL = +11.54 +/- 1.18 D (n = 13); +30 D IOL = +5.15 +/- 1.18 D (n = 105); +34.0 D IOL = +3.5 D (n = 1); +36 D IOL = +2.34 +/- 0.73 D 9 (n = 61); and +38 D IOL = +1.41 +/- 0.56 D (n = 28). Residual hyperopia ranged from +0.5 D to +2.5 D with +38 D IOL, and no eyes were myopic (overcorrected) by use of any of the IOL studied. Linear regression analysis of refractive state on IOL power for all dogs predicted that dioptric strength of +41.53 D was necessary to best approximate emmetropia for the population as a whole. Body type of the dog had only slight effect (< 1.0 D) on predicted optimal IOL power. Further linear regression analysis of the 7 breeds studied predicted variations from +39.62 to +43.14 D in IOL powers necessary to approximate emmetropia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427464 TI - Influence of body size on tibial nerve somatosensory evoked potentials in dogs. AB - Somatosensory evoked potentials in response to tibial nerve stimulation were recorded from the scalp of 31 clinically normal mixed-breed dogs. The latency and amplitude of a main positive potential (P18), recorded with a frontal electrode referenced to the nose, were measured in subjects with body length ranging from 316 to 962 mm. A linear relation to body size explained the variations in latency among dogs (r2 = 0.81); the amplitude variations were explained in part by body size (r2 = 0.44). Bilateral tibial nerve stimulation significantly (P < 0.05) increased the amplitude of P18, but its latency was unaffected, compared with unilateral stimulation. Results of unilateral right and left tibial nerve stimulation were compared and were not different. Replacing acepromazine with xylazine as premedication before thiopental anesthesia did not influence the recordings. PMID- 8427465 TI - Technique for long-term right dorsal colon fistulization in ponies. AB - Right dorsal colon fistulas, 2.5 cm in diameter, were created in 2 healthy ponies, using a 2-stage surgical procedure. The first stage consisted of resection of portions of the 16th and 17th ribs on the right side, followed by surgical creation of a 6- to 8-cm-diameter adhesion between the right dorsal colon and the body wall. Fistulas were created approximately 2 weeks after the first surgery by sharp dissection through the adhesion into the lumen of the colon. The fistulas have been satisfactorily maintained for > 2 years by de Pezzer catheters (45 F). Ponies with fistulas have been used for gastrointestinal experiments. PMID- 8427467 TI - Elbow arthroscopy in clinically normal dogs. AB - After a detailed anatomic study to determine puncture sites, 10 cadaver elbows from 5 dogs were examined arthroscopically to study the normal intraarticular anatomy, as viewed from the medial side. Subsequent dissection revealed absence of neurovascular injury and only minor iatrogenic damage to the cartilage. The technique was clinically applied and evaluated in 13 dogs (26 joints). The dogs recovered without complications. The technique proved to be safe and reliable for direct examination of nearly the entire joint. More specifically, it allowed systematic inspection of the medial and lateral humeral condyles, the medial and lateral coronoid processes, the caudal and middle parts of the head of the radius, the olecranon (including the anconeal process), and the medial collateral ligament. PMID- 8427466 TI - Electromyography of the pelvic diaphragm and anal sphincter in dogs with perineal hernia. AB - The innervation of the levator ani and coccygeal muscles and the external anal sphincter was studied by anatomic dissection in 6 clinically normal male dogs and by electrical stimulation in 5 clinically normal male dogs. Variations in innervation occasionally were found that were comparable to those reported in previous studies. Electromyographic recordings were made from the levator ani and coccygeal muscles and from the anal sphincter in 40 dogs during perineal hernia repair. Spontaneous potentials of 4 types were found in 35 dogs: fibrillation potentials, positive sharp waves, complex repetitive discharges, and fasciculations. Biopsy specimens of the cranial part of the levator ani muscle were taken in 12 dogs during perineal hernia repair. Histologic examination revealed atrophy in 7 specimens. Spontaneous potentials were recorded from all muscles with histologic evidence of atrophy. All examinations of the levator ani muscle concerned the cranial part of this muscle, because the caudal part was absent in all 40 dogs. From combined results of electromyography and histologic examination, it was concluded that atrophy of the muscles of the pelvic diaphragm, which develops in some dogs with perineal hernia, is likely to be of neurogenic origin. Nerve damage is localized in the sacral plexus proximal to the muscular branches of the pudendal nerve or in the muscular branches separately. PMID- 8427468 TI - Comparison of latex agglutination, indirect immunofluorescent antibody, and enzyme immunoassay methods for serodiagnosis of Rocky Mountain spotted fever in dogs. AB - Indirect immunofluorescent antibody (IFA), latex agglutination (LA), and enzyme immunoassay (EIA) methods were compared for evaluation of the serum antibody responses of dogs experimentally and naturally exposed to spotted fever-group rickettsiae. Selected sera (obtained on days 1, 42, 53, 124, 145, 236, 255, 264, and 292) were examined from three 8-month-old female Beagles inoculated with Rickettsia rickettsii on days 34 and 250 of the study. A second group of dogs comprised three 8-month-old female Beagles inoculated with R montana on days 34 and 102. Subsequently, these dogs were inoculated with R rickettsii on day 250. Serum samples were obtained from the second group of dogs on days 1, 96, 103, 132, 180, 215, 292, and 494. A third group consisted of 21 naturally exposed dogs, from which sequentially obtained serum samples were available, and which had clinical signs compatible with Rocky Mountain spotted fever. Clinical signs of disease in dogs of the third group resolved after treatment with tetracycline (22 mg/kg of body weight, PO, q 8 h) was instituted. At least 2 sequentially obtained serum samples from each dog were tested. In general, the first sample was obtained just prior to treatment and the convalescent serum samples were obtained at weekly or greater intervals thereafter. For correlation and reactivity data, an IFA test for IgG/IgM (using heavy and light chains-specific conjugate) was used as the reference standard for comparison of results with those of the other tests.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427469 TI - Evaluation of plasma cortisol and corticosterone responses to synthetic adrenocorticotropic hormone administration in ferrets. AB - Plasma cortisol and corticosterone responses of 8 clinically normal adult ferrets to synthetic ACTH (cosyntropin) were evaluated. Cosyntropin was administered IV at 4 dosages (0.5, 1.0, 5.0, and 10 micrograms/kg of body weight) at 2- to 4-week intervals, with blood samples collected 60 and 120 minutes after injection. After completion of the studies, an additional ACTH stimulation test was performed by administering cosyntropin (1.0 micrograms/kg) IM. The baseline plasma cortisol concentrations from all studies ranged from 25.9 to 235 nmol/L (mean +/- SEM = 73.8 +/- 7.0 nmol/L), and plasma corticosterone values ranged from 1.7 to 47 nmol/L (mean +/- SEM = 8.3 +/- 1.1 nmol/L). After IV administration of cosyntropin, plasma concentrations of cortisol and corticosterone increased significantly (P < or = 0.05) and reached peak values at 60 minutes; however, there were no significant differences between plasma cortisol or corticosterone responses to the 4 dosages of cosyntropin. Intramuscular administration of 1.0 micrograms of cosyntropin/kg induced increases in plasma cortisol and corticosterone concentrations that were similar to the responses induced by IV administration of cosyntropin. The mean molar ratio of cortisol to corticosterone, calculated from the resting plasma concentrations, was approximately 9:1, whereas the ACTH-stimulated cortisol to corticosterone ratio was approximately 4:1. Results of this study indicated that administration of cosyntropin to clinically normal ferrets, at dosages ranging from 0.5 to 10 micrograms/kg, increased plasma concentrations of cortisol and corticosterone. Although cosyntropin stimulates the adrenocortical secretion of cortisol and corticosterone, cortisol appears to be the predominate circulating glucocorticoid in ferrets. PMID- 8427470 TI - Changes in cross-sectional area and capillary supply of the muscle fiber population in equine gluteus medius muscle as a function of sampling depth. AB - The right and left gluteus medius muscles of 4 mature Andalusian stallions were examined by repeated needle biopsy over a specific area and depth, as well as at different depths, to determine whether the cross-sectional area and capillary supply of the various fiber types are homogeneous throughout the muscle. The muscle biopsy specimens were histochemically analyzed for fiber types (myofibrillar adenosine triphosphatase) and capillaries (amylase-periodic acid Schiff method). Differences between contralateral sites were not identifiable for any of the analyzed variables. Differences between sampling depths were larger than those between sample sites. The percentage of type-1 fibers increased, and that of type-2B fibers decreased as a function of increased sampling depth, but the relative frequency of type-2A fibers did not change with depth. The mean cross-sectional area of type-1 and type-2A fibers increased significantly, and that of type-2B fibers decreased between the most superficial and the deepest sampling sites. The capillary density and the mean number of capillaries per fiber increased as a function of sampling depth, but significant changes were not recorded between sampling depths for overall mean fiber area. The mean number of capillaries in contact with fibers of each histochemical type increased significantly with increasing sampling depth only for type-1 fibers. However, the number of capillaries in contact with fibers changed significantly, relative to the fiber area, as a function of sampling depth; for type-2B fibers this variable increased, whereas for type-2A and type-1 fibers, it decreased.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427471 TI - Qualitative and morphometric radiographic findings in the distal phalanx and digital soft tissues of sound thoroughbred racehorses. AB - Lameness examinations and radiography of the distal phalanx and associated soft tissue structures of the front feet of 103 Thoroughbred racehorses, 4 to 9 years old, were performed to determine normal radiographic appearance and morphometry. Of 103 horses examined, 41 were used in the study that were without clinical signs of foot problems or lameness, had raced at least twice prior to radiography, and had raced at least twice more in the 6 months after radiography. Lateromedial and dorsoproximal-palmarodistal radiographic views of each front distal phalanx were used to measure 28 bone and soft-tissue structures, and to evaluate 14 radiographic findings. Significant differences were not observed between left and right digits for any radiographic determination. Mean thickness of the soft tissues dorsal to the distal phalanx, which provides an evaluation of the epidermal laminae, was 14.6 +/- 1.0 mm when measured adjacent to the distal aspect of the distal phalanx. Most horses had straight, smooth hoof walls that were parallel to the dorsal cortex of the distal phalanx. The mean degree of palmar rotation of the distal phalanx was -0.5 +/- 1.3, and none was rotated more than 4 degrees. The dorsal cortex was smooth and straight, without bone deposition or reaction in either digit for only 5 of the 41 horses. Active bone formation was seen unilaterally along the middle portion of the dorsal cortex in 7 horses, and along the distal portion of the dorsal cortex in 4 of the phalanges from 3 horses. New bone formation along the distal dorsal cortex was often accompanied by resorption of the palmar cortex. For 26 of the 31 horses without active bone deposition, smooth inactive bone formation along the midportion of the dorsal cortex was identified in 1 or both distal phalanges. Bone at the solar margin of the distal phalanx was uniformly dense and finely trabeculated, without evidence of resorption or fractures. Severe irregularity of the solar margin was not found in any digit, and the margin of both phalanges was smooth in 8 horses. Various degrees of solar margin irregularity were observed in the other 33 horses. The mean number of vascular canals within the distal phalanx was 8.4 +/- 1.7, and the diameter of the largest canal was 3.4 +/- 0.6 mm.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8427472 TI - Serum triiodothyronine, total thyroxine, and free thyroxine concentrations in horses. AB - The objectives of this experiment were to determine serum concentrations of triiodothyronine (T3), thyroxine (T4), and free thyroxine (fT4) at rest, following thyroid-stimulating hormone (TSH) administration, and following phenylbutazone administration in healthy horses. This was done to determine which available laboratory test can best be used for diagnosis of hypothyroid conditions in horses. Serum T3, T4, and fT4 concentrations in serum samples obtained before and after TSH stimulation and following phenylbutazone administration for 7 days were determined. Baseline values ranged from 0.21 to 0.80 ng of T3/ml, 6.2 to 25.1 ng of T4/ml, and 0.07 to 0.47 ng of fT3/dl. After 5 IU of TSH was administered IV, serum T3 values increased to 6 times baseline values in 2 hours. Thyroxine values increased to 3 times baseline values at 4 hours and remained high at 6 hours. Free T4 values increased to 4 times baseline values at 4 hours and remained high at 6 hours. Administration of 4.4 mg of phenylbutazone/kg, every 12 hours for 7 days significantly decreased T4 and fT4 values, but did not significantly affect serum T3 concentrations. It was concluded that a TSH stimulation test should be performed when hypothyroidism is suspected. Measurement of serum fT4 concentrations, by the single-stage radioimmunoassay, does not provide any additional information about thyroid gland function over that gained by measuring T4 concentrations. Phenylbutazone given at a dosage of 4.4 mg/kg every 24 hours, for 7 days did significantly decrease resting T4 and fT4 concentrations, but did not significantly affect T3 concentrations in horses. PMID- 8427474 TI - Intraosseous pressure and pathologic changes in horses with navicular disease. AB - Navicular bone intraosseous pressure, gross pathologic, histologic, and histochemical data were collected from 8 horses with navicular disease and 4 control horses. Simultaneous navicular bone intraosseous, medial palmar arterial, and saphenous venous pressures were measured for the left and right forelimbs of each horse under general anesthesia. Gross pathologic evaluation included grading of changes on the flexor surface of the navicular bone. Safranin-O-fast green stained sections were used for histologic-histochemical grading of the hyaline articular and fibrocartilage surfaces of the navicular bones. Hematoxylin and eosin-stained sections were used for morphologic evaluation of the marrow spaces of navicular bones. Mean navicular bone intraosseous pressure for horses with navicular disease was significantly (P < 0.001) higher than that for controls. Differences in medical palmar arterial or saphenous venous pressures were not significant between groups. The median flexor surface gross pathologic and histologic-histochemical fibrocartilage scores for horses with navicular disease were significantly (P < 0.001) more severe than those for control horses. The histologic-histochemical hyaline cartilage scores for control horses and those for horses with navicular disease were not significantly different. Fibrosis of the marrow spaces beneath the flexor cortex of horses with navicular disease was more pronounced than that of control horses. PMID- 8427473 TI - Clinical pathologic profiles of dogs and turkeys with congestive heart failure, either noninduced or induced by rapid ventricular pacing, and turkeys with furazolidone toxicosis. AB - Characteristic alterations in the serum and urine biochemical profiles of Doberman Pinschers with congestive heart failure (CHF) resulting from idiopathic dilated cardiomyopathy were determined. We compared these alterations with those observed in 2 other models of CHF: rate overload induced by rapid ventricular pacing in dogs, and biventricular hypertrophy and dilatation induced in turkey poults by furazolidone toxicosis. Serum and urine biochemical changes in both models of CHF in dogs were mild to moderate in degree, and were moderately consistent. They could be attributed to secondary neurohumoral, hepatic, and renal effects of heart failure. The most marked and consistent changes observed were mildly decreased anion gap that developed, in part, because of decreased serum sodium concentration, moderately increased catecholamine concentrations, moderate lactaciduria, hyposthenuria, and mildly increased urea concentrations and liver enzyme activities. In birds with furazolidone cardiomyopathy, we observed mild increases in serum urate concentration, liver and muscle enzyme activities, but moderately increased sodium concentration with decreased chloride concentration. In the pacing and furazolidone models, in which CHF was rapidly induced, moderate to marked hypoproteinemia was attributable to decreases in albumin and globulin concentrations. Using the avian model we found that the hypoproteinemia could be largely attributed to blood volume expansion, and to a lesser extent, inanition. Development of hypoalbuminemia during rapid ventricular pacing and furazolidone treatment may contribute to the effects of rate overload or drug toxicity in the pathogenesis of CHF, because hypoalbuminemia may contribute to altered hemodynamics and neuroendocrine system activation. Our data indicate that clinical biochemical analysis of serum and urine may be useful for assessing progression of CHF. PMID- 8427475 TI - Type-I interferon genotypes and severity of clinical disease in cattle inoculated with bovine herpesvirus 1. AB - Genomic DNA samples and health records from 98 unrelated, mixed-breed cattle inoculated with bovine herpesvirus 1 (BHV-1) were examined to determine the relationship between interferon (IFN) genotype and severity of clinical disease. Cattle were retrospectively classified as moderately or severely affected on the basis of rectal temperature, feed intake, and weight gain after intranasal inoculation of BHV-1. Southern blot analysis of 16 type-I IFN genes identified alleles at 3 IFN loci (IFNB1, IFNW4, and IFNW8) that were significantly associated with the more severe clinical phenotype (odds ratios = 4.1 [P = 0.01], 2.3 [P < 0.05] and 2.4 [P = 0.06], respectively). A second allele at the IFNB1 locus was associated with the milder disease phenotype (odds ratio = 2.9, P < 0.05). These results indicate that selective breeding programs aimed at altering the frequency of these alleles in cattle populations may potentially improve animal health and lessen the economic impact of BHV-1 infection on cattle producers. PMID- 8427476 TI - Cytokine production during endotoxin-induced mastitis in lactating dairy cows. AB - The role of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor alpha during endotoxin-induced mastitis in cows was characterized. Six cows had 10 micrograms of Escherichia coli lipopolysaccharide infused into 1 mammary gland. Three other cows served as nontreated controls. Within 1.5 to 2.5 hours after infusion, endotoxin caused obvious edema of the mammary gland and increased serum albumin concentration in milk of infused glands 6 times. Milk somatic cell count began to increase 3 to 5 hours after infusion in all treated glands. At 7 hours after infusion, somatic cell counts were increased > 10 times, compared with counts in milk from control cows. Pyrexia of > 1 C developed in only 1 cow, but all treated cows had serum cortisol concentrations > 50 ng/ml in response to endotoxin treatment. High concentrations of IL-1 (10 to 600 U/ml) and IL-6 (2 to 22 U/ml) were detected in milk of infused glands beginning 2.5 to 4 hours after infusion. Endotoxin did not induce detectable amounts of tumor necrosis factor activity in milk or serum. Swelling and mammary gland permeability changes preceded any detectable increase in IL-1 and IL-6 activity, indicating that these clinical signs of inflammation were not mediated by these cytokines. Systemic responses and the leukocytic influx into endotoxin-infused glands developed after or concurrently with initial increases in IL-1 and IL-6 activities in milk. These results suggested that IL-1 and IL-6 may have a role in mammary gland defenses and in the pathophysiologic changes during endotoxin-induced mastitis. PMID- 8427477 TI - Enhancement of lymphocyte function and interleukin 1 beta transcription by recombinant bovine interleukin 1 beta. AB - Six nonpregnant, nonlactating Jersey cows, averaging 4 to 6 years old, were used to evaluate the immunomodulatory effects of recombinant bovine interleukin 1 beta (rBoIL-1 beta). Cows were given 166 ng of rBoIL-1 beta/kg of body weight at 8 hour intervals for 96 hours. Persistent leukocytosis was observed within 3 hours of rBoIL-1 treatment, peaking 24 hours after the first IL-1 beta injection and returning to baseline values within 72 hours after cessation of IL-1 beta treatment. Injection of cows with rBoIL-1 beta stimulated lymphocyte blastogenesis and mitochondrial methyl-thiazoltetrazolium cleavage activity in resting cell cultures. Increases in the aforementioned lymphocyte activities were also observed in stimulated blood mononuclear cell cultures during IL-1 beta administration. Change in IgM production in cell cultures was not observed during IL-1 beta treatment. Within 24 hours of the first IL-1 beta injection, IL-1 beta mRNA transcription in stimulated blood mononuclear cell cultures was markedly increased, suggesting that IL-1 beta upregulates its own production in mononuclear cells. These data provide evidence that administration of cytokines, such as rBoIL-1 beta, enhances immune cell function and, therefore, may be useful in alleviating immunosuppresion in cattle. PMID- 8427478 TI - Identification of Pasteurella haemolytica A1 isolates from market-stressed feeder calves by use of enzyme and antimicrobial susceptibility profiles. AB - An epidemiologic study of Pasteurella haemolytica serovar 1 (Ph1) in market stressed feeder calves from 7 farms in eastern Tennessee was conducted. The nasal mucus of each calf was cultured sequentially at the farm of origin (day 0), at an auction market (day 133), and at a feedyard in Texas (days 141, 148, 155, and 169). Of the 103 calves tested, 77 were culture-positive, including 1 on day 0, 1 on day 133, 20 on day 141, 57 on day 148, 50 on day 155, and 14 on day 169. From the 143 Ph1 isolates, 20 enzyme profiles were determined by use of a commercial enzyme system that detects 19 enzymatic reactions; 4 antimicrobial susceptibility profiles were obtained, using the disk-diffusion method, which evaluated susceptibility to 11 antibacterial drugs. All isolates were positive for acid phosphatase and alkaline phosphatase, but were negative for alpha-galactosidase, alpha-mannosidase, beta-glucosidase, beta-glucuronidase, cystine aminopeptidase, N-acetyl-beta-glucosaminidase, and trypsin. Other positive enzyme reactions included: leucine aminopeptidase, 140 Ph1 isolates; phosphohydrolase, 90 isolates; alpha-fucosidase, 63 isolates; esterase (C4), 59 isolates; valine aminopeptidase, 30 isolates; esterase lipase (C8), 24 isolates; beta galactosidase, 2 isolates; and alpha-glucosidase, chymotrypsin and lipase (C14), 1 isolate each. Thirty-four Ph1 profiles were identified, using combined enzyme and antimicrobial susceptibility profiles. The data indicate that the strains isolated during the feedyard period may have been determined more by farm of origin (P < or = 0.001) than by habitation with calves from other farms while in the feedyard.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427479 TI - Control of human aggression. A comparative perspective. AB - Recent animal research has demonstrated that humans are not a uniquely aggressive species and that even in so-called violence-prone animals, aggression is always an optional strategy. Although some form of intraspecific aggression exists in every vertebrate species studied thus far, it is also true that all organisms have coevolved equally potent inhibitory mechanisms that enable them to use an aggressive strategy selectively or to suppress aggression when it is in their interest to do so. Parallel studies of aggression in children, assaultive adults, and even entire societies have suggested that humans are exquisitely sensitive to subtle social controls that could be used to reduce the frequency of individual acts of violence. PMID- 8427480 TI - The structure of phenotypic personality traits. AB - This personal historical article traces the development of the Big-Five factor structure, whose growing acceptance by personality researchers has profoundly influenced the scientific study of individual differences. The roots of this taxonomy lie in the lexical hypothesis and the insights of Sir Francis Galton, the prescience of L. L. Thurstone, the legacy of Raymond B. Cattell, and the seminal analyses of Tupes and Christal. Paradoxically, the present popularity of this model owes much to its many critics, each of whom tried to replace it, but failed. In reaction, there have been a number of attempts to assimilate other models into the five-factor structure. Lately, some practical implications of the emerging consensus can be seen in such contexts as personnel selection and classification. PMID- 8427481 TI - Accuracy and objectivity on behalf of the APA. PMID- 8427482 TI - APA's expert panel in the congressional review of the USS Iowa incident. AB - In 1989, an explosion aboard the USS Iowa killed 47 sailors. The Navy attributed the explosion to the intentional suicidal acts of Gunners Mate Clayton Hartwig, a conclusion supported primarily by an "equivocal death analysis" conducted by the Federal Bureau of Investigation (FBI). The U.S. House of Representatives Armed Services Committee (HASC) was highly critical of the FBI's report and the Navy's conclusions, in part because of the peer review provided by 12 psychologists organized by the American Psychological Association (APA). This article (a) reviews the nature of equivocal death analysis and related reconstructive psychological evaluations, (b) describes the nature of APA's consultation and involvement with the HASC, (c) discusses the conclusions reached by the HASC and the influence of the APA panelists, and (d) suggests limitations on the use of equivocal death analysis and related procedures in light of scientific concerns and ethical considerations. PMID- 8427483 TI - Cisplatin and doxorubicin for invasive basal cell carcinoma of the eyelids. AB - Cisplatin and doxorubicin were used in the treatment of recurrent invasive basal cell carcinoma (BCC) of the medial canthus and orbit. Complete remission was observed after treatment termination, and this was still evident five years later. Systemic chemotherapy offers an alternative mode of treatment for selected cases of advanced BCC. PMID- 8427484 TI - Posttraumatic herpes zoster ophthalmicus as a presenting sign of human immunodeficiency virus infection. AB - We present the case of a 38-year-old man who developed herpes zoster ophthalmicus after orbital blunt trauma. Additional evaluation revealed human immunodeficiency virus type 1 (HIV-1) infection. This case shows that varicella-zoster may be activated by local trauma and that herpes zoster ophthalmicus in young patients may indicate underlying HIV-1 infection. PMID- 8427485 TI - Reduced variation of the diurnal curve after the subscleral scheie procedure for primary open-angle glaucoma. AB - We report the results of a one-year prospective study of the maximum intraocular pressure and the range of the oscillations of the diurnal curve after a subscleral Scheie procedure in 21 eyes (13 patients) with chronic primary open angle glaucoma. Pressure curves were recorded preoperatively, during the immediate postsurgical period, at one month postoperatively, and every three months thereafter up to one year after surgery. Our results showed that this procedure reduced the maximum intraocular pressure and the oscillations of the diurnal curve (P < .001). This effect was maximum during the immediate postoperative period and decreased one month after surgery (P < .001). PMID- 8427486 TI - Superior altitudinal hemianopia and herpes zoster. AB - A healthy 41-year-old women had acute retrobulbar optic neuritis with superior altitudinal hemianopia four weeks after cutaneous herpes zoster. Her visual acuity decreased to 0.04 OD, and mild iritis was noticed. However, ophthalmoscopic examination and fluorescein angiography disclosed no remarkable change. She had a low amplitude during flash visual-evoked potential testing with almost normal latency time in the acute stage. Corticosteroid therapy was administered, and her visual acuity and visual-field defect rapidly improved. Complete recovery, including the pattern-reversal visual-evoked potential results, was obtained. PMID- 8427487 TI - A comprehensive approach to biomicroscopic vitreous examination. AB - We recommend a biomicroscopic vitreous examination that combines an anterior vitreous examination done without using a neutralizing lens and a posterior vitreous examination using the high-power (low-magnification) double aspheric positive preset lens. When detailed observation of the posterior and peripheral vitreous are required, we also recommend the additional use of a low-power (high magnification) double aspheric preset lens and the Goldmann three-mirror lens. Details of the anterior vitreous can be examined best without using a neutralizing lens and those of the posteror vitreous, by using the low-power double aspheric preset lens. The high-power double aspheric preset lens provides a wider field of view and more accessibility when the patient has small pupils and cataracts than the low-power double aspheric preset lens. The peripheral vitreous is best observed using a Goldmann three-mirror lens. We believe the method we describe can serve as a guide for a comprehensive clinical biomicroscopic vitreous examination. PMID- 8427489 TI - Unilateral map-dot-fingerprint dystrophy after acute angle-closure glaucoma: a case report. PMID- 8427488 TI - Intraocular lymphoma invades the optic nerve and orbit. AB - A diagnosis uveitis OS was made in a 54-year-old woman; this was resistant to corticosteroid treatment. One year later, both eyes were involved. Examination of the vitreous aspirate and the results of a chorioretinal biopsy OD did not yield a correct diagnosis. Computed tomographic scanning showed orbital invasion OD. Malignant lymphoma was diagnosed after a biopsy specimen of the orbital contents was studied histopathologically. Radiotherapy and chemotherapy diminished the orbital mass and intraocular lesions. After regression of the orbital lesion, a brain tumor was found and treated with radiotherapy and chemotherapy. At age 57 years, the patient died. We believe that our patient had a rare case of intraocular lymphoma associated with orbital invasion. PMID- 8427490 TI - Capping device for sharp instruments. AB - Accidental needle puncture of the skin carries the risk of transmitting infection. Accidental needle-stick injuries comprise one third of work-related accidents. A capping device was devised to protect medical personnel during handling of sharp instruments. PMID- 8427491 TI - Mathematic proof of Schachar's hypothesis of accommodation. AB - To evaluate whether there is increased or decreased zonular tension during accommodation, we developed a qualitative small displacement model using Rayleigh's method to determine the characteristics of the human lens changes that occur during accommodation. We found that the optical power of the human lens increased linearly with zonular tension. Our results definitely prove Schachar's hypothesis of accommodation. PMID- 8427493 TI - Intracranial longitudinal splitting of facial nerve: a new approach for hemifacial spasm. AB - A new surgical technique for the treatment of hemifacial spasm consisting of longitudinal splitting of the facial nerve in the cerebellopontine angle is described. Thirty-three cases have been treated with good results. Follow-up of 20 cases for 1 year or more showed no recurrence or facial paresis. One case required a second operation. The novelty of this approach lies in its effectiveness in relieving the symptoms without running the risk of overmanipulating the aberrant artery. PMID- 8427492 TI - High jugular bulb: implications for posterior fossa neurotologic and cranial base surgery. AB - The suboccipital-retrosigmoid approach to the internal auditory canal and cerebellopontine angle is being used with increasing frequency for neurotologic surgery, including vestibular nerve section and resection of acoustic neuroma. It offers wide exposure of the cerebellopontine angle and the cranial nerve VII-VIII complex as it courses from the brain stem to the temporal bone. Exposure of the internal auditory canal can be achieved by removing its posterior bony wall. Safe utilization of this approach requires familiarity with the variable position of structures within the petrous bone, including the lateral venous sinus and jugular bulb. We report here a case in which bleeding resulted from injury to a high jugular bulb during surgical exposure of the internal auditory canal via the suboccipital route and discuss the regional anatomy of the jugular bulb based on study of 378 consecutive temporal bone specimens from the collection of the Massachusetts Eye and Ear Infirmary. High jugular bulb was defined as encroachment of the dome of the bulb within 2 mm of the floor of the internal auditory canal. Forty-six percent of scoreable specimens met this criterion. However, when donors less than 6 years of age were excluded, a high jugular bulb was identified in 63% of specimens. Relevance to neurotologic surgery of the posterior fossa is presented. PMID- 8427494 TI - Compliance of the patulous eustachian tube. AB - The compliance and ventilatory functions of the eustachian tube (ET) of 17 ears with patulous ETs were examined and compared to those of 16 ears with traumatic perforation of the eardrum (controls) by means of the tubal compliance test, the inflation-deflation test, and the forced response test. The tubal compliance was significantly lower in the patulous ETs than in the controls. An excessive patency of the ET was confirmed among the patients with patulous ETs by a low opening pressure during the inflation test and a low passive resistance on the forced response test. Active ventilatory dysfunction was also found to be significantly more common in the patulous ETs by the deflation test and the measure of dilation efficiency. These results indicated that the ET appeared to be rather rigid and less moveable when patulous than when normal. PMID- 8427495 TI - Immune response to Prevotella intermedia in patients with recurrent nonstreptococcal tonsillitis. AB - The role of three oral flora organisms (Prevotella intermedia, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) was investigated in 31 children with recurrent nonstreptococcal tonsillitis. Antibody titers to the three organisms were measured by enzyme-linked immunosorbent assay in the 31 patients, as well as in 32 control patients who had not suffered from recurrent tonsillitis. None of the individuals in either group suffered from periodontal or dental illness. Significantly higher antibody levels to P intermedia were found in the study group as compared to controls (median 91.0 versus 72.5; p = .02). In contrast, the antibody titers to the other two organisms were generally low (less than 0.30), and no difference was found among the two study groups. The elevated antibody levels to P intermedia, a known oral pathogen that is also isolated from most recurrently inflamed tonsils, suggest a pathogenic role for this organism in recurrent tonsillitis. PMID- 8427496 TI - Histopathology of olfactory mucosa in Kallmann's syndrome. AB - Olfactory mucosa was harvested by intranasal biopsy from a man with Kallmann's syndrome in whom the absence of the olfactory bulbs was documented by magnetic resonance imaging. On electron microscopic examination, several pathologic changes were evident in the olfactory mucosa. First, most olfactory neurons lacked cilia (ie, were morphologically immature). Second, the fila olfactoria had fewer than the normal number of axons, and a large proportion of them were apparently undergoing electron lucent degeneration. Finally, neuromatous collections of axons were seen superficial to the basement membrane in the epithelium. Similar changes have been observed in the mucosa of experimentally bulbectomized rodents. Accordingly, a constellation of pathologic changes--axonal degeneration, neuronal immaturity, and the formation of intraepithelial neuromas- seems to be characteristic of olfactory mucosa that cannot innervate the olfactory bulb in both humans and animals. On the basis of our observations, it is worth investigating the status of the olfactory bulb in other forms of human anosmia in which similar morphological changes are observed in the mucosa, such as persistent posttraumatic anosmia and isolated congenital anosmia. PMID- 8427497 TI - Effect of oxymetazoline on nasal and sinus mucosal blood flow in the rabbit as measured with laser-Doppler flowmetry. AB - The effect of topical oxymetazoline hydrochloride on the blood flow of the nasal and sinus mucosa of the rabbit was measured by laser-Doppler flowmetry. Oxymetazoline, the active component in clinically used nose drops, induced a dose dependent decrease of the nasal mucosal blood flow. This effect has previously been shown in humans and suggests the presence of alpha 2-adrenoceptors in the nasal mucosa of the rabbit. Doses of oxymetazoline used clinically in humans induced a 50% reduction of blood flow in rabbits. Rhythmic variations in blood flow were seen in 30% of the rabbits after administration of oxymetazoline. Additionally, oxymetazoline induced a dose-dependent decrease of the mucosal blood flow in the maxillary sinus when the drug was applied in the nose. A vasoconstricting effect of oxymetazoline on the arteries penetrating the maxillary sinus ostium is a possible explanation. This can have positive as well as negative consequences on acute sinus infections. PMID- 8427498 TI - Laser-Doppler measurements and electrocochleography during ischemia of the guinea pig cochlea: implications for hearing preservation in acoustic neuroma surgery. AB - Interruption of cochlear blood flow has been implicated as one of the causes of the sensorineural hearing loss that may occur during acoustic neuroma surgery. With the guinea pig as an animal model for cerebellopontine angle surgery, laser Doppler measurements were used to estimate the cochlear blood flow changes caused by compression of the eighth nerve complex. With compression, the laser-Doppler measurements decreased abruptly; somewhat later, the electrocochleographic potentials declined. When compression was released, laser-Doppler measurements usually returned immediately, followed later by return of the electrical potentials. Some of these potentials, including the compound action potential of the auditory nerve, often became transiently larger than their precompression values. Interposing bone between the laser-Doppler probe and the otic capsule, so that the total bone thickness approximated the thickness of the human otic capsule, decreased the laser-Doppler measurement, but changes caused by compression were still apparent. Thus, although the human otic capsule is much thicker than the guinea pig capsule, it may still be possible to make laser Doppler estimates of human cochlear blood flow. Laser-Doppler monitoring during acoustic neuroma surgery may be beneficial, because it could give earlier warning of ischemia than is currently available from electrocochleographic monitoring, thereby enabling earlier corrective action. Electrocochleography complements laser-Doppler measurements by indicating the physiologic state of the cochlea. PMID- 8427499 TI - Off-vertical axis rotational responses in patients with unilateral peripheral vestibular lesions. AB - Off-vertical axis rotation (OVAR) stimulates the otolith organs in a manner that is suitable for assessment of the otolith-ocular reflex. To further assess the potential clinical usefulness of OVAR, the eye movement responses of seven patients with surgically confirmed unilateral peripheral vestibular lesions were compared with the eye movement responses of a group of age-matched, healthy, asymptomatic control subjects. Patients and controls were tested with constant velocity rotations that followed a brief period of angular acceleration (velocity trapezoid) using either earth-vertical axis (EVA) rotation or OVAR. Both EVA and OVAR sinusoidal velocity profiles were also performed. Results indicated that each patient had 1) an asymmetric OVAR response, ie, a bias component whose direction was opposite normal when rotating toward the lesioned ear, and 2) a normal modulation component. Population data suggested that patients had 1) a more rapid decay of response than normal subjects during OVAR velocity trapezoids, 2) an increased phase lead as compared to normal subjects during sinusoidal OVAR, and 3) like normal subjects, a less rapid decay of response during OVAR velocity trapezoids than during EVA rotational velocity trapezoids. Taken together, these findings suggest that patients with unilateral peripheral vestibular deficits have abnormal otolith-ocular and semicircular canal-ocular reflexes but that a single labyrinth appears to provide an otolithic signal sufficient for qualitatively normal semicircular canal-otolith interaction. PMID- 8427500 TI - Experimental transferable vascular bed for laryngotracheal reconstruction: further observations. AB - Free fascia flaps have proven most reliable in providing a vascular bed for an epithelial free graft without adding bulk. They may be a useful tool for laryngotracheal reconstruction. A free flap consisting of a vascular network running in connective tissue can be developed on the rabbit external ear. The vascular characteristics of this flap were examined to test the reliability of the transferable vascular bed in laryngotracheal repair. The perichondrial free flap is useful for bringing an internal lining inside the lumen and for circumferential protection of supporting tissue. However, the natural tendency for surface contraction of perichondrium is a major disadvantage. PMID- 8427501 TI - Histamine metabolism in nasal polyps. AB - We attempted to determine the relationship of nasal polyps to histamine (HA) metabolism. Compared to that in allergy-related nasal polyps and infection related nasal polyps, the level of HA in aspirin-induced asthma-related polyps was significantly lower. Large differences between the groups were not observed in HA-synthesizing enzyme activity, but degradative enzyme activity was much higher in aspirin-induced asthma-related polyps than in other types of nasal polyps tested. These findings suggest the possibility that the amount of HA in polyps associated with aspirin-induced asthma was less because of greatly enhanced degradation. We found, in addition, that in nasal tissues such as polyps, histamine-N-methyl transferase, rather than histaminase, was the principal degradative enzyme. PMID- 8427502 TI - Ultrasound detection of anaerobic neck infection. PMID- 8427503 TI - Adenomatoid and calcifying epithelial odontogenic tumors. AB - The histologic diversity of the odontogenic epithelium's neoplasms is nicely illustrated by the adenomatoid and the calcifying epithelial odontogenic tumors. The former has a predilection for the maxilla, the latter for the mandible. Neither has a malignant variant, and each is controlled by conservative local removal. The recurrence rate of the adenomatoid odontogenic tumor is 0.2%, while the calcifying epithelial odontogenic tumor's rate is 14%. PMID- 8427504 TI - Endoscopic laser medial arytenoidectomy for airway management in bilateral laryngeal paralysis. AB - A review of our recent experience in patients with bilateral laryngeal paralysis is described. While we continue to use phrenic nerve transfers in patients with mobile arytenoids, patients with fixed arytenoids generally require some sort of vocal cord lateralization, either by arytenoidectomy and arytenoidopexy or by partial vocal cord resection. The endoscopic laser medial arytenoidectomy is a convenient and effective method for opening the posterior glottic airway. One arytenoid is reduced medially with the carbon dioxide laser. After about 3 months the opposite arytenoid can be treated similarly, if necessary. The procedure does not appear to affect arytenoid mobility, as the posterior commissure mucosa and underlying interarytenoid muscle are protected and hence unaffected by the procedure. Those patients with at least one mobile arytenoid cartilage are candidates for posterior cricoarytenoid muscle reinnervation. Although ansa cervicalis and phrenic nerve techniques have been described, the author has concentrated efforts on the phrenic nerve. This report describes the endoscopic laser medial arytenoidectomy procedure, while the phrenic nerve patients will be reported in a subsequent manuscript. PMID- 8427505 TI - Laser surgery for epithelial hyperplasia of the vocal fold. AB - A total of 88 patients with epithelial hyperplasia of the vocal fold were treated with endolaryngeal microscopic carbon dioxide laser surgery during the period from 1978 to 1990. The majority of the patients were male smokers. During the follow-up period ranging from 12 to 156 months, 15 patients developed recurrence(s). The recurrent lesion was benign epithelial hyperplasia in 8 patients and invasive carcinoma in 7 patients. The recurrences occurred only in preoperative smokers. Discontinuation of smoking following laser surgery did not affect the incidence of recurrence. The incidence of development of carcinoma did not differ between two histologic types: simple hyperplasia without cellular atypia and dysplasia associated with cellular atypia. The incidence of cancer development for dysplasia cases in the present series (6%) was lower than that in previous reports in which the carbon dioxide laser was not employed for the treatment. Postoperative improvement of vocal function was manifested in an increase of the maximum phonation time, a decrease of the mean airflow rate, and an increase in vibratory amplitude and mucosal wave. PMID- 8427506 TI - Correlation between the laryngeal brain stem evoked response and the laryngeal chemoreflex in the porcine model. AB - The laryngeal brain stem evoked response (LBR) represents the neural activity involved in laryngeal reflex pathways. The laryngeal chemoreflex (LCR) is a centrally mediated response consisting of apnea and hemodynamic changes that result from laryngeal stimulation. The purpose of this study is to determine the characteristics of the LBR that are predictive of LCR severity in the porcine model. The duration of apnea resulting from stimulation of the supraglottic larynx defined LCR severity. The LBR tracings were recorded from electrodes flanking the brain stem following direct electrical stimulation of the superior laryngeal nerve. The LBR peak latencies from piglets demonstrating prolonged LCR apnea were compared to those without an exaggerated LCR response. Two LBR peak latencies demonstrated a statistically significant difference between the two piglet groups. These peak latencies appear to be indicators of susceptibility to exaggerated laryngeal reflex sensitivity. Thus, the LBR may prove useful in identifying and evaluating subjects predisposed to conditions associated with dysfunctional laryngeal reflex activity. PMID- 8427507 TI - Where are peripheral analgesics acting? PMID- 8427508 TI - Significance of enzyme linked immunosorbent assay (ELISA) for antibodies to double stranded and single stranded DNA in patients with lupus nephritis: correlation with severity of renal histology. AB - The correlation between renal histology and class specific (IgG and IgM) antibodies to double stranded DNA (dsDNA) and single stranded DNA (ssDNA) was studied by enzyme linked immunosorbent assay (ELISA) in 40 untreated patients with systemic lupus erythematosus (SLE). The levels of IgG antibodies to dsDNA were significantly higher in patients with World Health Organisation class IV nephritis than in those with class I, class II, or class III nephritis. IgG antibodies to ssDNA were higher in patients with class IV than in those with class II nephritis. IgG antibodies to dsDNA showed a close correlation with the histological activity score and the amount of electron dense deposit. IgG antibodies to ssDNA showed only a weak correlation with the renal histological activity score. IgM antibodies to dsDNA and IgM antibodies to ssDNA were not correlated with renal histological features. Patients with moderate to severe nephritis had a lower ratio of IgM antibodies to dsDNA to IgG antibodies to dsDNA than those with mild nephritis. These results indicate that the measurement of IgG antibodies to dsDNA is predictive in evaluating renal histological activity in patients with SLE. PMID- 8427509 TI - Bone mineral density in patients with recent onset rheumatoid arthritis: influence of disease activity and functional capacity. AB - BACKGROUND: Generalised osteoporosis is often described in patients with rheumatoid arthritis (RA). The aim of this study was to evaluate disease related determinants of bone mineral density (BMD) in patients with RA. METHODS: Subjects were selected from a group of 147 patients with recent onset RA. Disease activity and functional capacity were studied prospectively in this cohort. Activity of the disease was assessed once every three months by various parameters, and functional capacity was measured with a health assessment questionnaire once every six months. Ninety seven patients consented to participate in the study. Bone mineral density was assessed with dual energy x ray absorptiometry in the lumbar spine, in a combined region of interest in the hips, and in Ward's triangle. Multiple linear regression procedures were used to analyse the data. RESULTS: Duration of RA was negatively associated with BMD at all three sites of measurement. The mean erythrocyte sedimentation rate in the six months before BMD measurement was negatively associated with BMD in the hip and in Ward's triangle. Other parameters of disease activity were not related to BMD. The mean health assessment questionnaire score in the 18 months before BMD measurement was negatively associated with BMD in the combined hip region only. Bone mineral density tended to be decreased when patients were compared with a normal reference group, especially in the femoral regions of interest. CONCLUSIONS: It is concluded that BMD may be affected in patients with recent onset RA by disease dependent mechanisms. Several factors have been suggested elsewhere as determinants of BMD in RA. The results of this study show that disease duration, disease activity, and functional impairment may, independently of each other, contribute to bone loss, especially in the proximal femur. PMID- 8427510 TI - Cartilage degradation by polymorphonuclear leucocytes: in vitro assessment of the pathogenic mechanisms. AB - Polymorphonuclear leucocytes (PMNs), which predominate in inflammatory synovial fluid, can degrade cartilage. This was measured by a novel in vitro model; PMNs were incubated for up to one hour with 2 or 3 microns sections of cartilage and the glycosaminoglycan loss determined by microdensitometry after alcian blue staining. Glycosaminoglycan loss could be as a result of damage from reactive oxygen species, proteolytic enzymes, or a combination of the two. The relative contributions of these mechanisms were evaluated using selective inhibitors. The results show that activated PMNs will degrade cartilage and that this degradation is due to proteolytic enzymes and not reactive oxygen species. There is a specificity involving elastase but not other serine proteases. It is suggested that enzyme inhibition may play a part in reducing PMN mediated cartilage damage. PMID- 8427511 TI - Synergism between muramyl dipeptide and lipopolysaccharide in the inhibition of glycosaminoglycan synthesis in cultured rat costal chondrocytes. AB - The effect of synthetic muramyl dipeptide on glycosaminoglycan synthesis in cultured rat costal chondrocytes was examined. Muramyl dipeptide alone had no effect on the glycosaminoglycan synthesis of rat chondrocytes, whereas Escherichia coli lipopolysaccharide and interleukin 1 alpha inhibited glycosaminoglycan synthesis in a dose dependent manner. Muramyl dipeptide, when added to chondrocyte cultures in the presence of lipopolysaccharide, enhanced the lipopolysaccharide induced inhibition of glycosaminoglycan synthesis in a dose dependent manner. Adjuvant active analogues of muramyl dipeptide, but not adjuvant inactive analogues, also enhanced the lipopolysaccharide induced inhibition of glycosaminoglycan synthesis. In combination with muramyl dipeptide, to inhibit glycosaminoglycan synthesis, lipopolysaccharide could be replaced with the synthetic lipid A, an active principle of lipopolysaccharide. These results show that the muramyl dipeptide portion of bacterial peptidoglycan enhances the susceptibility of rat chondrocytes to the lipid A portion of bacterial lipopolysaccharide, and therefore the interaction between chondrocytes and bacterial cell wall components might be involved in damaging the cartilage in inflammatory joint diseases. PMID- 8427512 TI - Inhibition of interleukin 1 beta induced rat and human cartilage degradation in vitro by the metalloproteinase inhibitor U27391. AB - Interleukin 1 induced proteoglycan loss from cartilage in vitro was prevented by a biochemical inhibitor of metalloproteinase activity. The inhibitor also partially relieved the inhibition of proteoglycan synthesis caused by interleukin 1. The loss of glycosaminoglycan by rat and human femoral head cartilage in response to human recombinant interleukin 1 beta (rhIL-1 beta) was established, and the modulation of this loss by the metalloproteinase inhibitor U27391 was investigated. Rat femoral head cartilage consistently lost glycosaminoglycan in response to rhIL-1 beta whereas only a proportion (30%) of normal human femoral head cartilage did so. Concentrations of 10-100 mumol/l U27391 inhibited the action of rhIL-1 beta on rat femoral head cartilage, reversing both the loss of glycosaminoglycan and the inhibition of glycosaminoglycan synthesis. U27391 also prevented the reduction in glycosaminoglycan content of those human femoral head cartilage explants responsive to rhIL-1 beta. Metalloproteinase inhibition therefore prevents rhIL-1 beta induced glycosaminoglycan loss by rat and human femoral head cartilage, suggesting that inhibitors of such enzymes may prove to be of therapeutic benefit in erosive diseases in humans. PMID- 8427513 TI - Musculoskeletal manifestations in hyperlipidaemia: a controlled study. AB - Eighty eight patients with hyperlipidaemia (81 white patients from South Africa and seven patients of mixed race from the West Cape area) were studied. Forty eight had adult familial hypercholesterolaemia, 16 had juvenile familial hypercholesterolaemia, and 24 had mixed hyperlipidaemia (increased cholesterol and triglycerides). They were interviewed and examined and their musculoskeletal manifestations compared with 88 controls with normal lipid profiles, and matched for age, sex, and race for each group of patients. The following manifestations were significantly increased in the patients: (a) tendon xanthomas particularly of the tendo Achillis in patients with adult familial hypercholesterolaemia and mixed hyperlipidaemia; (b) tendo Achillis tendinitis in patients with adult familial hypercholesterolaemia and mixed hyperlipidaemia; and (c) oligoarthritis in patients with mixed hyperlipidaemia but not in those with adult familial hypercholesterolaemia. Migratory polyarthritis and transient tendo Achillis pain were rare. Thirty eight per cent of patients with juvenile familial hypercholesterolaemia had musculoskeletal system manifestations none of which was significantly increased compared with controls. There was a significant association between tendon xanthomas and tendo Achillis tendinitis. There was a significant difference in pretreatment cholesterol levels in the patients with adult familial hypercholesterolaemia and musculoskeletal system manifestations compared with those without and in all three groups combined. The study confirms an association between hyperlipidaemia and tendon xanthomas, tendo Achillis tendinitis, and to a lesser extent oligoarthritis but not migratory polyarthritis or transient tendo Achillis pain as reported in other studies. It also shows that musculoskeletal system manifestations antedated the diagnosis of hyperlipidaemia in 24/39 (62%) patients and that the manifestations improved or resolved completely in 19/30 (63%) patients after receiving lipid lowering treatment. It is therefore important to recognise the association between musculoskeletal system manifestations and hyperlipidaemia for diagnostic and therapeutic reasons. PMID- 8427514 TI - Arthritis in the highlands of Papua New Guinea. AB - Acute polyarthritis is an important cause of morbidity in many tropical countries. Classification has often been difficult, with the term tropical polyarthritis used for those in whom a diagnosis could not be made. The implication that this is a distinct entity is probably incorrect, with likely causes being septic arthritis or post-infective reactive arthritis. This study aimed to determine the types of arthritis found in 43 patients (30 men) presenting consecutively to the Goroka Base Hospital in the Eastern Highlands of Papua New Guinea. Gonococcal arthritis was diagnosed in eight patients (six men) on the basis of isolation of Neisseria gonorrhoeae from the joint aspirate. In all cases the N gonorrhoeae was identified by the closed culture system on chocolate agar, but not always by routine plating. There were no specific clinical features that identified patients with a gonococcal septic arthritis. The remaining 34 patients had an undifferentiated oligoarthritis. The pattern of arthritis in men and women was of a lower limb pauciarticular arthritis with a predilection for the knee and ankle joints. A total of 30% of male patients had a history of urethral discharge and 44% of all patients had preceding diarrhoea. Arthritis was the only feature in 59% of patients and in 32% there was an associated enthesitis. In this study most patients had an oligoarthritis consistent with a reactive arthritis or a septic arthritis due to N gonorrhoeae. Broth inoculation of synovial fluid was the best method to isolate N gonorrhoeae, with standard methods for gonococcal isolation failing in some patients. It is recommended that the term 'tropical polyarthritis' is no longer used as it does not refer to a specific entity but consists of several known arthritides. PMID- 8427515 TI - Paget's disease of bone treated with a five day course of oral tiludronate. AB - Chloro-4-phenyl thiomethylene bisphosphonate (tiludronate) is a new drug which can be used as an inhibitor of bone resorption. As it remains in bone for a long time, and as mineralisation defects have only been seen at doses much higher than those required to decrease osteoclastic activity, it could be given at high doses over a short period of time. Eighteen patients with Paget's disease of bone were randomly allocated to three therapeutic groups receiving respectively 600, 800, and 1200 mg/day tiludronate for five days. Serum alkaline phosphatase activity and the urinary hydroxyproline/creatinine ratio were quickly and drastically reduced in all three groups. A significant reduction of serum alkaline phosphatases and the hydroxyproline/creatinine ratio was still present six months after the five day therapeutic course, reflecting a sustained activity of tiludronate even after stopping treatment. Dose dependent short and long term reductions of bone turnover rate were observed. Biochemical assessment of haematological, renal, or hepatic tolerance did not show any toxicity of tiludronate. Fifty per cent of patients treated by a dose of 1200 mg/day reported gastrointestinal disturbances, however, making this dosage unsuitable for clinical practice. PMID- 8427516 TI - Impact of disablement due to rheumatic disorders in a British population: estimates of severity and prevalence from the Calderdale Rheumatic Disablement Survey. AB - A survey of rheumatic disablement in the population has enabled the comparative impact of self reported causes of disability to be studied. One in three households in Calderdale, West Yorkshire, United Kingdom was screened in 1986 with a postal questionnaire (87% response rate), followed by in depth interviews with a sample of subjects reporting disability in conjunction with a rheumatic disorder (608 interviews). Severity of disablement was assessed using the physical independence handicap classification. The estimated prevalence of disability in conjunction with reported rheumatic disorders is 82/1000 population aged at least 16 years (95% confidence interval (CI) 77 to 87). Arthritis (mainly osteoarthritis) is the most commonly reported cause (47/1000 population; 95% CI 43 to 51), followed by back or neck disorders (25/1000; 95% CI 23 to 28), soft tissue disorders (18/1000; 95% CI 15 to 20), and rheumatoid arthritis (RA) (4/1000; 95% CI 3 to 5). A total of 30% reported more than one category of rheumatic disorder (mean number 1.3) and 63% reported non-rheumatic comorbidity. Current joint symptoms were reported by 98%, current antirheumatic drugs (including analgesics) by 70%, and severe pain by 62%. Overall 82% of subjects had seen their general practitioner in the past year, and 71% reported having attended an outpatient clinic; 26% reported current outpatient clinic attendance, and 15% a hospital inpatient stay during the previous year. Forty six per cent reported some dependence, with 12% reporting being dependent on a daily basis. Rheumatoid arthritis was the most disabling disorder with 73% dependent. Taking into account prevalence, osteoarthritis and back disorders are the most, and RA the least, common causes of dependence and incapacitating pain in the population. This challenges stereotypes and raises questions about the organisation and priorities for specialist services and for research. PMID- 8427517 TI - Cutaneous vasculitis and IgA glomerulonephritis in ankylosing spondylitis. AB - Two patients with ankylosing spondylitis were found to have IgA nephropathy and leucocytoclastic cutaneous vasculitis. Immunofluorescence showed perivascular deposition of IgA in the skin of one patient and in the mesangium of both patients. Such an association has been reported only once before. This supports the concept of abnormal IgA immune stimulation in the pathogenesis of ankylosing spondylitis. PMID- 8427518 TI - Acquired Brown's syndrome in a patient with systemic lupus erythematosus. AB - A 27 year old woman with systemic lupus erythematosus (SLE) developed vertical diplopia with an apparent bilateral inferior oblique muscle palsy, resulting in a limitation of elevation of the globe in adduction. It resolved with systemic steroid treatment. A transient tenosynovitis affecting the superior oblique tendons was the probable underlying pathological mechanism. This is the first described case of Brown's syndrome associated with SLE. PMID- 8427519 TI - Menstrual arthritis. AB - The menstrual cycle is characterised by variations in the absolute and relative concentrations of the hormones of the hypothalamic pituitary ovarian axis, which in turn affect cell function and cytokine and heat shock protein production. Menstruation involves the shedding of the secretory endometrium, which is part of the mucosal associated lymphoid tissue and hence is rich in immunologically competent cells such as CD8 T cells and macrophages. The case is reported here of a patient presenting with a recurrent but transient symmetrical inflammatory polyarthritis which only occurred at menstruation with no residual damage. The disease was suppressed by danazol. Endometrial degradation products are suggested as the trigger of this 'menstrual arthritis'. PMID- 8427520 TI - Iron in joint inflammation. PMID- 8427521 TI - That oestrogen replacement for osteoporosis prevention should no longer be a bone of contention. PMID- 8427522 TI - Arthritis in hyperimmunoglobulinaemia D. PMID- 8427523 TI - Use of cyclosporin A in the eosinophilia myalgia syndrome. PMID- 8427524 TI - Postural variation in von Willebrand factor antigen. PMID- 8427525 TI - Hypertrophic osteoarthropathy and AIDS. PMID- 8427526 TI - Disseminated gonococcal infection in an elderly patient. PMID- 8427527 TI - Vacuum sign in spondylodiscitis due to H aphrophilus. PMID- 8427528 TI - Idiopathic orbital myositis: treatment with cyclosporin. PMID- 8427529 TI - Non-Hodgkin's lymphoma: initial manifestation. PMID- 8427530 TI - Arthritis and carcinoma. PMID- 8427531 TI - Endothelium damage and von Willebrand factor antigen. PMID- 8427532 TI - Food intolerance in rheumatoid arthritis. PMID- 8427533 TI - Insulin, insulin resistance, and blood pressure elevation. PMID- 8427534 TI - Erythropoietin. Biology and clinical applications. AB - Hemopoiesis is a complex process that underlies the production of highly specialized cells. The mechanisms involved in this process include positive and negative feedback by humoral activities, pluripotent stem cell self-renewal and differentiation, and local interactions between stromal components of the hemopoietic microenvironment and various stem and progenitor cells. PMID- 8427535 TI - The empathic physician. AB - Empathy is a process for understanding an individual's subjective experiences by vicariously sharing that experience while maintaining an observant stance. It is a useful tool in the medical encounter as it provides the physician with a fuller, more personalized view of the patient, and it provides the patient with a sense of connectedness to the physician that may allow him/her to more freely express his/her emotional distress. The roots of empathy are explained as a process that evolves from a developmental substrate with the addition of relevant experience, memory, and fantasy. While understanding the patient alone is a worthwhile goal, the physician's empathic insight can have therapeutic impact by its reflection back on the patient, through the use of language, to express support or sympathy, to justify behavior, or to foster deeper emotional expression. PMID- 8427536 TI - HIV-infected health care professionals. Public opinion about testing, disclosing, and switching. AB - BACKGROUND: We wanted to know what the public believes about the risks of human immunodeficiency virus (HIV) transmission in health care settings, and what opinions the public holds regarding HIV-infected health care professionals. We also wanted to uncover the correlates and predictors of those opinions. METHODS: A telephone survey of a nationwide random probability sample of adults was conducted in summer 1991. Thirteen hundred fifty adults completed the survey. The response rate was approximately 63%. We assessed (1) public opinion about whether HIV-infected physicians, surgeons, and dentists should quit working, and (2) the public's self-reported intention to remain in the care of an HIV-infected professional or to switch to another provider. RESULTS: Public concern about HIV transmission in health care settings has increased from 19% in 1988 to 38% in 1991. More of the public now believes that transmission from HIV-infected physicians is likely (up from 33% in 1988 to 46% in 1991). Yet, fewer respondents believe that HIV-infected physicians should not be allowed to work (45% vs 39%). Only 5% would deprive HIV-infected physicians of their livelihood as physicians. Fewer would switch from HIV-infected physicians now than in 1988 (56% vs 37%). Knowing someone with HIV infection was related to less concern and to less belief in likelihood of transmission as well as to increased support of HIV-infected health professionals' right to work. CONCLUSIONS: Although the public is more concerned about HIV transmission in health care settings since 1988, fewer would not allow HIV-infected health care professionals to work now than in 1988. Personalizing the epidemic, by using personal physicians and people with acquired immunodeficiency syndrome as educators, might help continue the trend toward improved attitudes toward HIV-infected health care professionals. PMID- 8427537 TI - Association of fasting insulin with blood pressure in young individuals. The Bogalusa Heart Study. AB - BACKGROUND: The relationship between fasting plasma insulin and blood pressure was studied in a cross-sectional survey of children and young adults aged 5 to 26 years. METHODS: Fasting plasma insulin, glucose, blood pressure, and anthropometric measurements were obtained on 3518 individuals. RESULTS: When divided into four age groups, the analyses showed that fasting insulin was significantly and positively correlated to both systolic and diastolic blood pressure in individuals at all age groups, except at 13 to 17 years. In multivariate regression analyses, fasting insulin remained independently associated with blood pressure levels after controlling for glucose levels, body mass index (weight/height) and skinfold thickness in children (aged 5 to 12 years) and young adults (aged 18 to 26 years), although not in adolescents (aged 13 to 17 years). Moreover, fasting insulin was more strongly related to systolic than to diastolic blood pressure. The fasting blood glucose level did not contribute independently to multivariate prediction of blood pressure in young adults. When the children and young adults were divided into tertiles according to fasting insulin and body mass index, the independent effect of insulin and body mass index on systolic pressure was also seen in children and young adults. CONCLUSIONS: The association between plasma insulin and blood pressure noted even in healthy children and young adults help target areas for cardiovascular risk prevention. PMID- 8427538 TI - Hereditary nephropathy associated with hyperuricemia and gout. AB - BACKGROUND: The clinical characteristics of hereditary nephropathy associated with hyperuricemia or gout have not been fully described, and the pathogenetic role of increased serum urate concentration is controversial. METHODS: We examined the clinical characteristics of 14 patients and purine metabolism of seven patients, while they were on a purine-restricted diet, in two families with hereditary nephropathy associated with asymptomatic hyperuricemia or gout. Results of plasma and urinary purine measurements were compared with those obtained in 25 patients with gout and renal insufficiency and in 25 normal subjects. Eight subjects in both families were followed up for a mean of 44 months. Allopurinol was given to all patients and enalapril maleate to hypertensive subjects. RESULTS: All patients had some combination of hyperuricemia, gout, renal insufficiency, arterial hypertension, and reduced kidney size. Decreased glomerular filtration rate was proportional to the decreased renal plasma flow. Renal vascular resistance was markedly increased in the patients with diminished renal plasma flow. All patients with familial nephropathy showed diminished urinary uric acid, hypoxanthine, and xanthine excretion rates. Purine under-excretion was more severe in affected patients with familial nephropathy than in patients with gout and renal insufficiency. Kidney biopsy specimens from three patients with familial nephropathy showed tubulointerstitial lesions and ischemic changes in glomeruli but no uric acid crystals. The kidney uric acid content was normal. Allopurinol treatment normalized serum urate levels, but serum creatinine concentrations increased and creatinine clearance decreased in all patients with familial nephropathy. One patient with gout only at initial evaluation developed renal failure during the follow-up period. CONCLUSIONS: Increased serum urate concentrations in hereditary nephropathy associated with hyperuricemia and gout are due to severe impairment of uric acid excretion. Hyperuricemia does not appear, however, to be of pathogenetic relevance and may be a consequence of a primary disruption of renal hemodynamics. PMID- 8427539 TI - Clarithromycin and other antimicrobial agents in the treatment of disseminated Mycobacterium avium infections in patients with acquired immunodeficiency syndrome. AB - BACKGROUND: Disseminated infection with Mycobacterium avium is common with late stage acquired immunodeficiency syndrome (AIDS), and no antimicrobial agent has been found to be clearly effective. METHODS: A multicenter open trial was conducted to assess the antimicrobial activity and clinical efficacy of clarithromycin--a new macrolide antibiotic--against disseminated M avium in 77 patients with late-stage AIDS. Blood cultures were taken at baseline and during treatment; side effects were also evaluated. RESULTS: Mycobacterium avium was eradicated from blood cultures in 11 (63%) of 16 evaluable patients receiving daily doses or 500 or 1000 mg, (n = 21) and in 45 of 46 (98%) of those receiving 1500 or 2000 mg (n = 56). Eradication after 2 months was influenced by continuity of drug treatment; 36 of 42 patients with no relapse had received continuous treatment vs six of 14 patients whose drug treatment had been stopped for 7 days or longer. After 2 to 7 months of treatment, acquired resistance associated with relapse was observed. Drug side effects were elevated liver enzyme levels (26%) and impaired hearing (4%). Concomitant AIDS drugs had no favorable effect on outcome and may have worsened liver toxicity. CONCLUSIONS: Clarithromycin has bacteriologic efficacy against M avium infection in late-stage AIDS, although drug resistance eventually develops. Further studies are needed to investigate safe, effective concomitant drugs. PMID- 8427540 TI - Advance directives. Effect of type of directive on physicians' therapeutic decisions. AB - BACKGROUND: Despite growing support for advance directives, there are few data validating their utility. We conducted this study to determine if the type of advance directive influences physicians' willingness to withhold specific therapies and if physicians are equally willing to withhold these therapies. METHODS: The 444 full-time faculty of the department of medicine of a university medical center were presented with patient scenarios and accompanying advance directives in three separate surveys that were mailed. They were asked if they would withhold each of 12 specific therapies based on their interpretation of the advance directive. Three types of advance directives were studied: general statement, therapy specific, and therapy specific combined with designation of a proxy and prior patient-physician discussion. RESULTS: The mean proportion of all 12 therapies that were withheld varied by type of advance directive: general statement advance directive, 55%; therapy-specific advance directive, 71%; and therapy-specific advance directive with proxy and prior patient-physician discussion, 83%. Respondents were more likely to withhold cardiopulmonary resuscitation when given a therapy-specific advance directive vs general statement advance directive, 84% vs 73%, respectively. With a therapy-specific advance directive that was supported by a proxy and prior patient-physician discussion, 100% of physicians were willing to withhold cardiopulmonary resuscitation. With the therapy-specific advance directive that was supported by proxy and prior patient-physician discussion, physicians were not equally willing to withhold all therapies, ie, mechanically assisted ventilation, 98%; intravenous fluids, 82%; antibiotics, 80%; simple tests, 70%; and pain medications, 13%. CONCLUSIONS: Detailed advance directives with a supportive proxy, coupled with physician-patient discussion, furnish the most reliable medical directives. Even with such directives, physicians are more likely to withhold life-saving therapies than simple tests, treatments, and pain medications. PMID- 8427541 TI - Deceptively normal ventricular fluid in lymphomatous meningitis. AB - The diagnosis of leptomeningeal cancer ultimately depends on the finding of abnormal cerebrospinal fluid with malignant cytologic study results. We report a case of relapsed leptomeningeal lymphomatosis in which ventricular cerebrospinal fluid was entirely normal while lumbar spinal fluid was diagnostically abnormal. To our knowledge, this is the first such reported case, and it highlights the importance of sampling cerebrospinal fluid close to the site of clinical involvement. PMID- 8427542 TI - Pneumocystis carinii thyroiditis. Report of three cases and review of the literature. AB - Pneumocystis carinii infection of the thyroid gland has previously been described in only four living patients with acquired immunodeficiency syndrome, three of whom had been receiving inhaled pentamidine prophylaxis against P carinii pneumonia. We treated three additional patients with P carinii thyroid involvement, all of whom were receiving aerosolized pentamidine. Two of our patients presented with clinical features suggestive of subacute granulomatous thyroiditis. The diagnosis of P carinii in our patients, as well as in the previously described patients, was established by thyroid fine-needle aspiration and Gomori's silver methenamine stains. The recent emergence of P carinii infection of the thyroid gland is likely related to the use of inhaled pentamidine prophylaxis, which appears to predispose to the development of extrapulmonary pneumocystosis. Clinicians need to be aware of the possibility of P carinii thyroiditis and should use aspiration and Gomori's methenamine silver staining in studying patients with the acquired immunodeficiency syndrome who have a painful (or other unexplained) thyroid mass so as to be able to initiate prompt and appropriate therapy. PMID- 8427543 TI - Dehydrating to terminate is different from dehydrating the terminal. PMID- 8427544 TI - Admission serum albumin and prognosis in acquired immunodeficiency syndrome. PMID- 8427545 TI - Human immunodeficiency virus-associated autonomic neuropathy, drug addiction, and zidovudine treatment. PMID- 8427546 TI - Copper in nuts may lower heart disease risk. PMID- 8427548 TI - Induction of extracellular arabinases on monomeric substrates in Aspergillus niger. AB - The induction of extracellular arabinases by pentose sugars and polyols generated by the metabolic pathway of L-arabinose and D-xylose catabolism in Aspergillus niger was investigated. Induction occurred with L-arabinose and L-arabitol but not with D-xylose or xylitol. L-arabitol in particular was found to be a good inducer for alpha-L-arabinofuranosidase and endo-arabinase activities. Western blotting analysis showed both alpha-L-arabinofuranosidase A and B to be present. No induction was observed using D-arabitol. Unlike the wild type A. niger N402 strain, the A. niger xylulose kinase negative mutant N572 also showed induction of alpha-L-arabinofuranosidases A and B and endo-arabinase activity on D-xylose and xylitol. This is due to metabolic conversion of these compounds leading to the accumulation of both xylitol and L-arabitol in this mutant, the latter of which then acts as inducer. The induction of the two alpha-L-arabinofuranosidases and endo-arabinase is under the control of two regulatory systems namely pathway specific induction and carbon catabolite repression. Under derepressing conditions in the wild type only alpha-L-arabinofuranosidase B could be detected by Western blotting analysis. This indicates that alpha-L-arabinofuranosidase B is of importance in the initiation of specific induction of the various arabinose activities in A. niger grown on arabinose containing structural polysaccharides. PMID- 8427547 TI - Biosynthesis of cyclic beta-(1-3),beta-(1-6) glucan in Bradyrhizobium spp. AB - Inner membranes of Bradyrhizobium japonicum strain USDA 110 produced in vitro soluble and insoluble beta-(1-3),beta-(1-6) glucans. The reaction proceeded through a 90 kDa inner membrane intermediate protein; used UDP-glucose as sugar donor and required Mg2+. Gel chromatography of soluble glucans resolved a cyclic beta-(1-3) glucan with a degree of polymerization of eleven from a family of beta (1-3),beta-(1-6) glucans with variable degree of polymerization higher than eleven. Bradyrhizobium strains BR4406 and BR8404 isolated from tree legume nodules in Southeast Brazil produce beta-(1-3), beta-(1-6) glucans very similar to that of B. japonicum. A 100 kDa protein was identified in these strains as intermediates in the synthesis of these glucans. Inner membranes of B. japonicum USDA110, B. japonicum I17, and Bradyrhizobium strains BR4406 and BR8404 incubated with UDP-glucose were unable to synthesize beta-(1-2) glucan and lacked the 235 kDa intermediate protein known to be involved in the synthesis of beta-(1-2) glucan in Agrobacterium tumefaciens, Rhizobium meliloti and Rhizobium loti. PMID- 8427550 TI - Congruence of service utilization estimates from the Epidemiologic Catchment Area Project and other sources. AB - Service utilization estimates for inpatient and ambulatory mental health care from the Epidemiologic Catchment Area Project were compared with similar estimates from other sources, principally the Center for Mental Health Services National Reporting Program. Generally, results showed closer correspondence between estimates of the number of persons who used inpatient care than of similar estimates for ambulatory mental health care. Subtotal estimates for the specialty alcohol/other drug abuse/mental health and health care sectors were more similar than were estimates for individual settings. The specialty sector subtotals showed only a 7% difference in patient counts for inpatient care and 13% for ambulatory care, with an 11% difference in visits for the latter. Generally, a reasonable level of congruence was observed, given pronounced differences in methods, procedures, and instruments. Future directions may be able to close data gaps and improve the quality of the national mental health services database. PMID- 8427549 TI - Induction of SOS like responses by nitrofurantoin in Vibrio cholerae el tor cells. AB - Treatment of Vibrio cholerae el tor strain SLH22(J) with nitrofurantoin induced dose-dependent prophage 'kappa', the maximum induction being 6-fold the spontaneous induction level. UV-inactivated 'kappa' phages were Weigle reactivated, the maximum Weigle factor being 1.8 and 2.0 respectively in nitrofurantoin and UV pretreated el tor strain H218 Smr. Nitrofurantoin treatment also caused significant filamentation of the el tor strain H218 Smr and mutation of these cells from ampicillin sensitivity to ampicillin resistance. The levels of the four SOS-like responses induced by this drug were low but significant. PMID- 8427551 TI - Progress toward achieving a common language in psychiatry. Results from the field trial of the clinical guidelines accompanying the WHO classification of mental and behavioral disorders in ICD-10. AB - In preparing for the 10th revision of the International Classification of Diseases (ICD-10), the Division of Mental Health of the World Health Organization organized an international field trial to help evaluate draft clinical descriptions and diagnostic guidelines that were produced to facilitate use of the chapter dealing with mental and behavioral disorders. These clinical guidelines were prepared in equivalent versions in most of the world's widely spoken languages. The field trial aimed to obtain data that would help in assessing whether the classification fits the diagnoses made in different countries, whether it is easy to use, and whether psychiatrists after a short period of familiarization with the classification can reach agreement about their diagnoses and classification. The field trial was carried out at 112 clinical centers in 39 countries by 711 clinicians who conducted 15,302 individual assessments. The trial included joint clinical assessments of patients and case history exercises. The results of the joint assessment part of the trial are reported here. Most clinicians reported that the draft document was easy to use and that the classification provided a good fit for the vast majority of the clinical conditions encountered. While interrater reliability was satisfactory for most categories, some (for example, those dealing with personality disorders) were somewhat difficult to use, and reliability of assignment for those was lower. The trial demonstrated that the ICD-10 chapter dealing with mental and behavioral disorders is on the whole suitable for general use. It provided valuable indications about changes needed for subsequent versions and demonstrated the feasibility of large-scale international research on classification and diagnosis in psychiatry. PMID- 8427552 TI - The Shatin community mental health survey in Hong Kong. II. Major findings. AB - A large-scale community survey in Shatin, Hong Kong, is presented with a modified two-phase design using flagged and nonflagged subsamples. A modified Self Reporting Questionnaire and the Diagnostic Interview Schedule (version III) were used as the screening and diagnostic instruments, respectively. Lifetime rates for 19 Diagnostic Interview Schedule/DSM-III diagnoses are presented. The most common diagnoses in Shatin were tobacco dependence, generalized anxiety disorder, alcohol abuse and/or dependence, all phobias, and dysthymic disorder. The male predominant disorders were tobacco dependence, alcohol abuse/dependence, pathological gambling, and antisocial personality. The female-predominant disorders were generalized anxiety disorder, all phobias, dysthymic disorder, major depressive disorder, obsessive-compulsive disorder, and bereavement. PMID- 8427553 TI - Suicide among women related to number of children in marriage. AB - The hypothesis of a negative association between rates of suicide and number of children in marriage was investigated in a prospective study of 989,949 women followed up for 15 years (1970 through 1985) with 1190 deaths from suicide. Women who had never married exhibited higher relative risks for suicide than married parous and married nonparous women for all age groups younger than 65 years at the start of follow-up. Among the married, the parous women had lower relative risks than nonparous women for all ages. For both premenopausal and postmenopausal women, a strong linear decrease in relative risk for suicide with increasing number of children in marriage was found. The effect of number of children was independent of social class measured as years of completed schooling. The findings provide the first empirical support for theories of parenthood and suicide advanced by Durkheim almost 100 years ago. PMID- 8427554 TI - Death without warning? A clinical postmortem study of suicide in 43 Israeli adolescent males. AB - Forty-three consecutive Israeli male suicides, 18 to 21 years of age, that occurred during compulsory military service were studied using preinduction assessment data, service records, and extensive postmortem interviews with family and peers. At preinduction, subjects, as a group, appeared above average in intelligence, physical fitness, and measures predictive of successful adaptation to military service. Active duty performance was generally satisfactory. Ascertained post mortem, 53.5% met formal criteria for major depressive disorder; most cases, however, appeared recent and reactive. Narcissistic and/or schizoid traits were common. Substance abuse was absent and antisocial personality disorder was rare (4.7%). Furthermore, in eight patients (18.6%) no Axis I diagnosis could be made; half of these also lacked any significant Axis II pathology. These findings, at partial variance with US studies, suggest a complex relationship between suicide and mental disorder. The striking failure of intensive screening and preventive measures to prevent these suicides highlights unresolved questions of etiology and intervention. PMID- 8427555 TI - Suicide attempts in patients with panic disorder. AB - In a clinical sample of 100 outpatients with panic disorder, 42% had a history of suicide attempt. Female sex and being single, divorced, or widowed were associated with an increased risk of suicide attempt. Thirty-one (73.8%) of the suicide attempts occurred after the first panic attack and 27 (64.3%) after the onset of panic disorder. Eighty-eight of these patients met DSM-III-R criteria for at least one other diagnosis in addition to panic disorder. Moreover, 52% had a history of major depressive episode and 31% had a lifetime diagnosis of alcohol and/or other substance abuse. Compared with those who did not attempt suicide, those who attempted suicide were significantly more likely to have suffered from major depressive episode and alcohol or other substance abuse in their lifetime. Among the 35 patients with no comorbidity with either major depression or addictive behaviors, 17.1% had a history of suicide attempt. All had suffered from depressive symptoms and/or a personality disorder. The same association was found in four of 19 patients suffering from panic disorder only with or without agoraphobia. PMID- 8427556 TI - Pregnancy/delivery complications and psychiatric diagnosis. A prospective study. AB - We examined the hypothesis that pregnancy and delivery complications result in increased risk for the development of psychiatric disorders. The study sample included 1068 pregnancies classified as chronic fetal hypoxia, other complications, preterm birth, or normal pregnancy/delivery that had initially been studied prospectively from the prenatal period through age 7 years. Subjects were recontacted (ages 18 to 27 years) and lifetime psychiatric diagnoses made with the Diagnostic Interview Schedule. Preterm subjects had significantly higher rates of cognitive impairment. Subjects with chronic fetal hypoxia had higher rates of both cognitive impairment and psychotic disorders, although these differences failed to reach statistical significance due to the small number of cases. With these exceptions, the data did not support the hypothesis that rates of psychiatric disorders are higher among subjects born with complications of pregnancy and delivery than among normal controls born without complications. PMID- 8427557 TI - Impact of treatment accessibility on clinical course of parasuicidal patients. PMID- 8427559 TI - The legal definition of professionalism in medicine. PMID- 8427558 TI - The de facto US mental and addictive disorders service system. Epidemiologic catchment area prospective 1-year prevalence rates of disorders and services. AB - After initial interviews with 20,291 adults in the National Institute of Mental Health Epidemiologic Catchment Area Program, we estimated prospective 1-year prevalence and service use rates of mental and addictive disorders in the US population. An annual prevalence rate of 28.1% was found for these disorders, composed of a 1-month point prevalence of 15.7% (at wave 1) and a 1-year incidence of new or recurrent disorders identified in 12.3% of the population at wave 2. During the 1-year follow-up period, 6.6% of the total sample developed one or more new disorders after being assessed as having no previous lifetime diagnosis at wave 1. An additional 5.7% of the population, with a history of some previous disorder at wave 1, had an acute relapse or suffered from a new disorder in 1 year. Irrespective of diagnosis, 14.7% of the US population in 1 year reported use of services in one or more component sectors of the de facto US mental and addictive service system. With some overlap between sectors, specialists in mental and addictive disorders provided treatment to 5.9% of the US population, 6.4% sought such services from general medical physicians, 3.0% sought these services from other human service professionals, and 4.1% turned to the voluntary support sector for such care. Of those persons with any disorder, only 28.5% (8.0 per 100 population) sought mental health/addictive services. Persons with specific disorders varied in the proportion who used services, from a high of more than 60% for somatization, schizophrenia, and bipolar disorders to a low of less than 25% for addictive disorders and severe cognitive impairment. Applications of these descriptive data to US health care system reform options are considered in the context of other variables that will determine national health policy. PMID- 8427560 TI - The 1990 Comprehensive Blood Bank Surveys of the College of American Pathologists. AB - The performance of participants on the 1990 Comprehensive Blood Bank Surveys of the College of American Pathologists was consistent with the high quality of past surveys. The surveys include challenges in ABO and Rh typing, crossmatching, transfusion decisions, unexpected antibody detection and identification, and supplemental questions to assess contemporary practices. As in previous years, a recurrent serologic problem has been the reporting of antibodies not present in serum (anti-E in Set J-B, and anti-K in Set J-D). Such overidentification of antibodies has little implication for transfusion safety and may, in fact, represent an artifact of the survey's samples and reporting practices. PMID- 8427561 TI - The diagnosis of Alzheimer's disease. PMID- 8427562 TI - Making the diagnosis of Alzheimer's disease. A primer for practicing pathologists. AB - There is demand on community pathologists to perform autopsies to confirm the clinical diagnosis of Alzheimer's disease, the most common cause of dementia in our increasingly aging society. Yet many pathologists are reluctant to examine autopsy brains because they have little experience with neuropathology and with the common histopathologic staining methods needed to evaluate dementia cases. This article provides interested pathologists with a simple, practical protocol to use in meeting this demand. While there is no absolute diagnostic gold standard for Alzheimer's disease and the histopathologic diagnosis remains imperfect, the guidelines presented are adapted from those used by many neuropathologists at Alzheimer's disease research centers participating in CERAD, the Consortium to Establish a Registry for Alzheimer's Disease. Recipes for appropriate stains and specific case examples are provided for convenience. PMID- 8427563 TI - Hepatic inflammatory lesions manifested as a pseudotumor. Report of two cases with different characteristics. AB - We report two surgical cases of inflammatory conditions of the liver that, on imaging studies, closely resembled a neoplasm. Under the diagnoses of hepatic malignant neoplasms, both patients underwent hepatic resections. Histologically, one case was found to be a hepatic xanthogranuloma (an inflammatory pseudotumor) that consisted of foamy histiocytes, plasma cells, lymphocytes, and fibroblasts, while the other case was found to be an epithelioid cell granuloma with a central area of coagulative necrosis. Although their exact pathogenesis remained obscure, cholangitis in the former case and tuberculosis in the latter were assumed to be possible causal factors. Special attention should be paid to these unusual conditions in terms of the differential diagnosis of hepatic mass lesions. PMID- 8427564 TI - Biliary cystadenocarcinoma arising in a liver with fibropolycystic disease. AB - Intrahepatic cholangiocarcinoma associated with fibropolycystic disease of the liver and biliary cystadenocarcinoma are rare tumors that are considered distinct entities. We present a case of a malignant tumor with features of hepatic cystadenocarcinoma arising in a background of fibropolycystic disease. PMID- 8427565 TI - Acid phosphatase activity in senile plaques and cerebrospinal fluid of patients with Alzheimer's disease. AB - The origin of the various components of senile plaques in Alzheimer's disease (AD) continues to be a focus of intense research scrutiny. Lysosomal enzyme activity within the plaques is of particular interest because of its possible relevance to a presumed abnormal processing of precursor molecules that may lead to the formation of plaque amyloid. Histochemical evidence of acid phosphatase (ACP), a lysosomal hydrolase, activity in senile plaques has been documented long before many of the current biochemical data regarding plaque pathogenesis became available. Recent evidence suggests the presence of neuronal membrane abnormalities that may allow "leakage" of some intracellular molecules, including enzymes, into the perineuronal environment and from there, possibly, to the cerebrospinal fluid (CSF). In the course of investigating this hypothesis, we decided to readdress the issue of ACP activity in AD by assaying the enzyme both in brain tissue sections as well as in CSF samples from neuropathologically confirmed cases of AD and from normal control subjects. Brain frozen sections from five cases with AD and five control subjects were histochemically stained for ACP using the alpha-naphthylphosphate hydrolysis method. Frozen CSF samples from 15 cases with AD and 19 control subjects were assayed for ACP activity using the thymolphthalein monophosphate hydrolysis method. In all cases studied, CSFs were cytologically unremarkable. Neurons in normal and AD brains were strongly positive for ACP. In AD, numerous senile plaques showed strong ACP activity with both granular and diffuse patterns. The CSFs from six of 15 AD cases (40%) showed ACP activity with values ranging from 0.04 to 0.4 U/L. No ACP activity was detected in any of the 19 control CSFs analyzed. The exact source of ACP in senile plaques and CSF of patients with AD cannot be established based on these data alone but the ACP may have originated from neurons with oxidative stress or oxygen free radical-mediated membrane damage. The data encourage further investigation of this hypothesis. PMID- 8427566 TI - Localized hypertrophic mononeuropathy of the trigeminal nerve. AB - We report a case of localized hypertrophic mononeuropathy that involved the trigeminal nerve in a 61-year-old woman with a 5-year history of progressive drooping of the left eyelid, double vision, and left-sided temporal and retro orbital pain. A mass that occupied the patient's left cavernous sinus was found to display localized hypertrophic mononeuropathy characterized by "onion bulbs" that were formed of concentrically proliferating Schwann cell processes around intact axons. This contrasted to the majority of previously reported cases of localized hypertrophic mononeuropathy, predominantly found in peripheral nerves, where the proliferating cell processes have been shown to be of a perineurial cell origin. Differences in architectural arrangement and degree of cellularity between the perineurial and schwannian forms of localized hypertrophic mononeuropathy were noted. These findings suggest important fundamental differences in the pathogenesis of various forms of onion-bulb mononeuropathies. PMID- 8427567 TI - Quality assurance study of cardiac isoenzyme utilization in a large teaching hospital. AB - Guidelines for diagnosis of acute myocardial infarction recommend that, if acute myocardial infarction is suspected, creatine kinase (CK)-MB levels should be measured on admission and again at 12 and 24 hours. In light of these recommendations, we conducted a quality assurance study to determine whether utilization of CK-MB tests in our institution, a large, university-affiliated teaching hospital, was consistent with current guidelines. Also, several years ago, we had established a policy of cancelling lactate dehydrogenase isoenzyme orders if the request originated from an unauthorized location, unless it was approved by a laboratory staff. Since this policy led to a greater than 90% reduction in the requests for lactate dehydrogenase isoenzyme testing, an additional objective was to reevaluate this policy. Of 774 patients evaluated with CK-MB tests, 294 (38%) received only a single test. Of these single tests, 277 had normal results (CK-MB < 5%). For the remaining 17 patients, the single CK MB test findings were abnormal (CK-MB > 5%) without follow-up testing. Only two CK-MB tests were ordered for 187 patients (24%). Three or more CK-MB tests were obtained in 293 cases (38%). When two or more CK-MB tests were ordered, the time interval between the first and second tests was inappropriately short in 70% and long in 24%. The recommended timing for the third CK-MB was followed in only 4% of cases. Review of 32 cancellations of lactate dehydrogenase isoenzyme tests disclosed that lactate dehydrogenase isoenzyme tests were requested when unnecessary in 26 cases. Despite published guidelines for use of CK-MB for acute myocardial infarction, physicians at our institution continue to use these tests inappropriately by ordering only single CK-MB tests or by ordering repetitions of CK-MB tests at excessively short or long intervals. PMID- 8427568 TI - Determination of malnutrition in hospitalized patients with the use of a group based reference. AB - We clustered a selected population of patients at Bridgeport (Conn) Hospital into distinct nutritional classes using critical decisions points for the serum concentrations of aspartate aminotransferase activity, cholesterol, total protein, albumin, and prealbumin. The decision points that divided the populations with the highest efficiency were delineated by the information in the data set. At these decision values the hospital population adjusted malnutrition rate was just more than 30%. This was lower than the reported prevalence of malnutrition in the hospital population and was better in agreement with global estimates of malnutrition. The study identified a problem in the assignment of decision values that are used for nutritional support which has implications for nutritional interventions. PMID- 8427569 TI - Factors associated with serum alkaline phosphatase level. AB - Serum alkaline phosphatase was measured in a large population of government employees and their dependents, aged 12 years and older. It was found that the concentrations of this enzyme were very high in early adolescence, fell to a low point after the completion of bone growth, and rose thereafter. The concentration was higher in males than females until the age of 50 years; after that it was higher in females. In employees and their spouses (aged 19 years and older), the greater the weight (or weight for height), the higher the concentration. The taller the person, the lower the level of this enzyme. The more cigarettes smoked per day, the higher the enzyme concentration. There was also a weak but statistically significant association of alkaline phosphatase with serum glucose concentrations and blood pressure but not with antihypertensive medications. Other factors were also considered. Some of these factors, particularly age and weight, are associated with substantial difference in the concentration of alkaline phosphatase and could conceivably affect the clinical interpretation of test results. PMID- 8427570 TI - Microcracks in the calcified layer of articular cartilage. AB - Evidence is presented that basophilic radial hairlines in the calcified layer of articular cartilage are in vivo microcracks. They were present in three of 25 normal patellas from subjects below the age of 50 years, in 20 of 25 recently fractured hips (average patient age, 77.3 years), and in 16 of 25 osteoarthritic tibial plateaus (71.7 years). It is hypothesized that extension of these microfractures beyond the calcified layer mediates remodeling of the osteochondral junction in aging and degenerative joint disease. PMID- 8427571 TI - Amyloid-producing squamous cell carcinoma of the uterine cervix. AB - We describe two unusual cases of squamous cell carcinoma of the uterine cervix associated with prominent intratumoral deposition of amyloid. Most of the amyloid is deposited within and between the tumor islands, but some amyloid globules are intracellular. The amyloid shows immunoreactivity for keratin, suggesting that it is derived from cytokeratin intermediate filaments, similar to the pathogenesis of primary cutaneous amyloidosis. Tumor-related amyloidosis is a heterogeneous but distinctive subgroup of amyloidosis, including cytokeratin-derived amyloid deposited within squamous or squamous-related tumors as in the present cases, polypeptide-derived amyloid associated with endocrine tumors, and systemic inflammation-associated amyloid (AA protein) as observed rarely in Hodgkin's disease and renal cell carcinoma. PMID- 8427572 TI - Ciliated cell remnants in peritoneal dialysis fluid. AB - Ciliocytophthoria are anucleate remnants of ciliated epithelial cells derived from the lower respiratory tract and female reproductive tract. We report a case of ciliocytophthoria found in the effluent dialysis fluid of a young woman undergoing long-term ambulatory peritoneal dialysis. Inability to identify these "organisms" initially led to an extensive search for parasitic contamination or infection of the peritoneum. After identifying these "organisms" as ciliocytophthoria, a prospective study showed that ciliated cell remnants occur frequently in the effluent dialysate of young women, but not in older women or men. With increasing use of peritoneal dialysis, laboratory personnel can expect to see ciliocytophthoria in peritoneal dialysate effluent and should recognize them as benign, normal findings in young women. PMID- 8427573 TI - Multiple plasmacytomas in skin. Harbinger of aggressive B-immunocytic malignancy. AB - A 55-year-old alcoholic man with chronic atypical mycobacterial lung disease developed multiple plasmacytomas in the skin over a 6-month period. There was no evidence of either multiple myeloma or plasma cell leukemia at the time of diagnosis; however, evidence of multiple myeloma became apparent about 7 months later, and the man died, despite therapy for this condition, about 22 months after the initial diagnosis of his plasma cell disorder. Despite the apparent rarity of this clinical presentation, the available evidence suggests it is a harbinger of a relatively aggressive B-immunocytic malignancy. PMID- 8427574 TI - Benign schwannoma of the liver. AB - We report a case of benign schwannoma presenting as a rapidly enlarging liver mass in a 67-year-old man and review the literature on nerve sheath tumors of the hepatobiliary system. Such tumors, which may occur in patients with and without neurofibromatosis, are very uncommon in the liver. The case reported herein, to our knowledge, is the first well-documented benign schwannoma of the liver occurring in a patient without neurofibromatosis. PMID- 8427575 TI - Greater than the sum of its parts. II. Matter-energy processing subsystems. AB - Behavioral science theory has changed greatly since 1978, when Miller's Living Systems was published. Much of the change represents progress, particularly in the biological fields. During that time, living systems theory (LST) developed in many ways. Among the most significant developments have been the addition of another level, the community, to the seven levels previously included in LST and the addition of another subsystem, the timer, to become the 20th subsystem. The present article is the second to be published in this journal which describe relatively briefly and simply the current state of scientific knowledge about the subsystems of living systems. It deals with the first six matter-energy processing subsystems, the ingestor, distributor, convertor, producer, matter energy storage, and extruder. PMID- 8427576 TI - N-linked oligosaccharides of simian immunodeficiency virus envelope glycoproteins are dispensable for the interaction with the CD4 receptor. AB - In contrast to others groups, we have previously shown that N-linked glycans of HIV-1 and HIV-2 envelope glycoproteins do not play a major role in the gp-CD4 interaction. In order to clarify these inconsistencies, we investigated the role of N-glycans in the interaction of SIV with the CD4 receptor. The inhibition of binding of radiolabeled SIV envelope glycoprotein (gp140) to CD4+ cells by increasing concentrations of soluble CD4 shows that the interaction occurred with high affinity (K0.5 = 1.4 x 10(-8) M). Treating SIV gp140 with endo F-N glycanase, with or without detergent, reduced its molecular mass from 140 to 75 KDa. This indicates that N-glycan represents about 50% of molecular mass of the glycoprotein. Interestingly, the fact that deglycosylated SIV gp140 as its native form bound significantly and specifically to CD4 receptor indicates that oligosaccharides of SIV gp140 are not required for the gp-CD4 interaction. PMID- 8427577 TI - Immunoregulatory functions of paf-acether. IX. Modulation of apoptosis in an immature T cell line. AB - We investigated the ability of a phospholipid cytokine, paf-acether to modulate programmed cell death in an immature human T cell line CEM-C12. Paf-acether alone did not cause cell death, but when it was added to CEM-C12 cells in the presence of calcium ionophore, a marked increase in cell mortality and DNA fragmentation was observed compared with calcium ionophore alone. This effect was dose dependent between 2 and 10 microM paf-acether and specific in that lysophosphatidylcholine had a minimal effect. Thus, in association with another signal, paf modulates apoptotic processes in an immature human T cell line. This may be relevant to intrathymic lymphocyte maturation. PMID- 8427578 TI - A DNA polymerase alpha-associated 56 kDa protein kinase. AB - Initiation of chromosomal DNA replication must be carefully regulated during the cell cycle. We report that Drosophila embryo DNA polymerase alpha complex, isolated by immunological techniques, contains a protein kinase activity. The kinase will phosphorylate histone H1, but prefers peptides contained in the DNA polymerase alpha-kinase complex. Renaturation experiments determined that the kinase activity resides in a 56-kDa peptide. PMID- 8427579 TI - Molecular characterization of the 5'-flanking region of human genomic ETA gene. AB - A clone containing the promoter, the exon 1 and exon 2 of human genomic ETA gene was isolated and sequenced. The transcription initiation site and TATA box were identified at 528 bp upstream of the initiation codon and 24 bp upstream of the transcription initiation site, respectively. The 534 bp fragment containing 5' flanking sequence and most of the exon 1 of the human ETA gene showed a promoter activity corresponding to 55% of SV40 early promoter activity when placed upstream of the luciferase gene and transfected into CHO cells. Thus, the TATA box which consists of the TAAAAA sequence is functional for transcription of the human ETA gene. The coding region of the human ETA receptor, therefore, starts from exon 2 which contained the first and the second transmembrane regions. The human ETA gene was demonstrated to be a single pair of copy by Southern blot analysis, and it was localized in chromosome 4 by analysis with a panel of human rodent somatic cell hybrid lines. PMID- 8427580 TI - The effect of epinephrine on the intracellular free calcium of parent and Ki-ras transfected NIH3T3 cells. AB - Here we describe differences in the formation of the epinephrine-induced Ca2+ transients between parent and Ki-ras-transformed NIH3T3 fibroblasts. These transients proved to be the results of an efflux from both the thapsigargin sensitive and -insensitive intracellular pools. While the epinephrine-induced detachment of the ras-transformed cells might be due to cytoskeletal and/or cell matrix alterations. PMID- 8427581 TI - DNA binding and cleavage of a novel antitumor antibiotic dynemicin A: detection of binding and cleavage sites by exonuclease III digestion and alkali-induced cutting reactions. AB - We found here that dynemicin A effectively breaks DNA strands under alkaline pH condition. Binding of dynemicin A to double-stranded DNA clearly interrupts the digestion reaction of exonuclease III. The DNA association sites of dynemicin A correspond considerably well to its cleavage sites. Dynemicin A seems to intercalate preferentially into the relaxed region of the DNA double helix. On the other hand, the alkali-product of dynemicin A was chromatographically identified with dynemicin N, suggesting a DNA cleavage mechanism similar to the reductant- and light-induced activation systems of dynemicin A. In order to detect drug binding to the relaxation structure in the DNA duplex, the present exonuclease III-digestion-stop-sequence method is useful. PMID- 8427582 TI - Partial nucleotide sequence of the Xanthomonas maltophilia chorionic gonadotropin like receptor. AB - Xanthomonas maltophilia possesses a unique high-affinity binding site which binds human chorionic gonadotropin (hCG), but not human luteinizing hormone (hLH) or other glycoprotein hormones. We designed primers from the known nucleotide sequence of the human LH/CG receptor, spanning an area extending from transmembrane region 2 to transmembrane region 6. Genomic DNA extracted from Xanthomonas maltophilia was used to obtain a PCR amplified product using the above primers. The primary amplification product was cloned in a pCR11 TA cloning vector, and the partial nucleotide sequence of the gene determined. This determined sequence showed 73% identity with the human, as well as the rat LH/CG receptor. Comparison of the translated protein sequence with the human, rat and porcine receptor protein sequences showed a 52% similarity. PMID- 8427583 TI - 5'-RACE PCR of mRNA for three taxon-specific crystallins: for each gene one promoter controls both lens and non-lens expression. AB - Gene recruitment of enzyme crystallins is a novel process in molecular evolution in which genes for some metabolic enzymes acquire extremely high expression in lens without prior gene duplication. Using the RACE method, the 5' end of the mRNAs for duck lens delta 1-, argininosuccinate lyase/delta 2- and lactate dehydrogenase-B/epsilon-crystallin has been amplified from lens and non-lens tissues and sequenced. In all three cases the major transcription start sites were identical in lens and in other tissues. This suggests that for these three genes, and in contrast to at least one other example, gene recruitment does not require the presence of alternative tissue-preferred promoters. PMID- 8427584 TI - Characterization of the promoter of human prostaglandin H synthase-1 gene. AB - The gene of human prostaglandin H synthase-1 (PGHS-1,EC 1.14.99.1) was isolated from a human cosmid library and was contained in two overlapping clones. Multiple transcriptional start sites were identified. The major one located 136 bases upstream from the ATG initiation codon. The promoter region contains no canonical TATA box but possesses a high G+C content. These observations suggest that PGHS-1 gene has the characteristics of a housekeeping gene. Both 0.8 and 0.4 kb of the 5'-flanking sequence of the PGHS-1 gene can confer transcriptional activity. PMID- 8427585 TI - Vascular pericytes not only regulate growth, but also preserve prostacyclin producing ability and protect against lipid peroxide-induced injury of co cultured endothelial cells. AB - Interactions between vascular pericytes and endothelial cells were studied. Bovine retinal pericytes were found not only to inhibit growth of co-cultured human umbilical endothelial cells, but also to cause significant stimulation of prostacyclin production in the endothelial cells, both of which were achieved by close apposition of the two cell types. Substitution of a fibroblast feeder layer for the pericytes gave no change. Moreover, pericytes could confer protection against lipid peroxide-induced injury of endothelial cells, as determined by the release of [3H]-2-deoxy-D-glucose from radiolabeled endothelial cells. These results indicate that pericytes provide instructive signals to adjacent endothelial cells, helping to maintain microvascular homeostasis. PMID- 8427586 TI - Transcriptional attenuation of PIM-1 gene. AB - Levels of the mRNA for PIM-1, a protooncogene encoding a cytoplasmic serine threonine kinase show a wide variation among tissues and cell lines, although this gene is transcribed from a GC- rich housekeeping promoter. Previous studies have failed to identify tissue specific elements in the PIM-1 promoter raising the possibility that these elements might reside within the gene. Transient transfections of Luciferase reporter gene constructs into the chronic myelogenous leukemia cell line K562 (which expresses high levels of PIM-1 mRNA) demonstrate that the 1.7kbp PIM-1 promoter sequences alone were three times more efficient than constructs driven by the promoter+PIM-1 genomic sequences. Nuclear run on assays of nascent RNA from K562 cells revealed premature transcriptional termination within the PIM-1 gene. Thus, PIM-1 gene may be constitutively transcribed in all tissues and transcriptional attenuation could be one of the mechanisms regulating the observed differences in steady state levels of mRNA. PMID- 8427587 TI - Insulin accelerates the post germinative development of several fat-storing seeds. AB - The effect of insulin and insulin like growth factors I and II on sunflower, watermelon and cucumber cotyledons has been examined. Each peptide stimulates an increase in the activity of several glyoxysomal enzymes which catalyze the conversion of fat to carbohydrate. These results provide the first evidence for the action of insulin and insulin like growth factors in plants. PMID- 8427588 TI - Protease activity of botulinum neurotoxin type E and its light chain: cleavage of actin. AB - We demonstrate here for the first time a proteolytic activity of botulinum neurotoxin type E which is not expressed unless the single chain approximately 150 kDa neurotoxic protein is nicked into the dichain approximately 150 kDa neurotoxin. Actin was cleaved, in vitro, at multiple sites by the dichain neurotoxin and the N-terminal approximately 50 kDa light chain segment isolated from the dichain neurotoxin. The scissile peptide bonds of actin invariably contained Arg or Lys at the P1 site. Proteolytic activity of the isolated light chain and expression of this activity in the dichain form of the neurotoxin are consistent with the light chain's and the neurotoxin's intracellular actions- inhibition of neurotransmitter release. PMID- 8427589 TI - Molecular cloning, functional expression and mRNA analysis of human beta adrenergic receptor kinase 2. AB - In the present study the cDNA of human beta ARK2 was cloned using both PCR and cDNA library screening, subcloned into an expression vector and transiently expressed in COS7 cells. The expressed kinase activity was approximately 40% as efficient as human beta ARK1 in phosphorylating bovine rod outer segments in vitro. Northern blot analysis of human and bovine mRNA revealed a species specific pattern of multiple hybridization bands, with two major transcripts in human rather than one in bovine. High levels of mRNA expression were found in peripheral blood leukocytes. PMID- 8427590 TI - Characterization and expression of a novel human fatty acid-binding protein: the epidermal type (E-FABP). AB - Using PAGE--Autoradioblotting technique we have characterized an E--FABP in human epidermal cells that is distinct from liver-, heart-, intestine- and adipose tissue-FABPs. FABP radiobinding analysis was performed directly on protein extracts without prior partial purification. E-FABP has a Mr of approximately 15 kDa and binds oleic acid with high affinity but does not bind all-trans-, 13-cis- and 9-cis-retinoic acid nor all-trans-retinol. Expression levels of E-FABP were low in normal epidermis, higher in human cultured keratinocytes and still higher in psoriasis, a disease characterized by abnormal epidermal differentiation. These findings suggest that epidermal cells may have a distinct fatty acid metabolism compared to other tissues. PMID- 8427591 TI - The role of amino-terminal disulfide bonds in the structure and assembly of human fibrinogen. AB - Human fibrinogen contains two half-molecules, each composed of an alpha, beta, and gamma chain linked by disulfide bonds. The two half-molecules (alpha beta gamma) are held together in the native protein by additional disulfide bonds located in the amino terminus of each chain. Site-directed mutagenesis, in which the amino-terminal Cys residues (alpha-Cys28 and 36, beta-Cys65, and gamma-Cys8 and 9) were converted to Ser, was carried out in order to study the role of the amino-terminal disulfide bonds in the structure and assembly of fibrinogen. An analysis of the fibrinogen synthesized in transfected baby hamster kidney (BHK) cells employing various combinations of these mutations revealed that alpha-Cys36 and beta-Cys65 form disulfide bonds between two alpha beta gamma half-molecules, rather than within the same half-molecule; furthermore, these two disulfide bonds are sufficient to hold the two alpha beta gamma half-molecules together as intact fibrinogen. Disulfide bonds formed by gamma-Cys8 and 9 were also sufficient to hold the two fibrinogen alpha beta gamma half-molecules together, while the disulfide bond between the two alpha-Cys28 residues failed to form in the absence of the disulfide bonds linking the alpha and beta chains and the two gamma chains. PMID- 8427592 TI - Stimulation of a Gs-like G protein in the osteoclast inhibits bone resorption but enhances tartrate-resistant acid phosphatase secretion. AB - Previous studies have demonstrated that G-protein agonists induce quiescence (Q effect) or retraction (R effect) in isolated osteoclasts. We now report the functional effects of such agonists on osteoclastic bone resorption and enzyme release. Exposure of osteoclasts to tetrafluoro-aluminate anions (AlF4-), a universal G protein stimulator, resulted in a marked concentration-dependent inhibition of bone resorption. This was associated with a dramatic increase in the secretion of the osteoclast-specific enzyme, tartrate-resistant acid phosphatase (TRAP). Cholera toxin, a Gs stimulator and a selective Q effect agonist, similarly abolished bone resorption and enhanced TRAP secretion. In contrast, pertussis toxin, a Gi inhibitor and a selective R effect agonist, inhibited bone resorption significantly, but slightly reduced enzyme release. The results suggest an involvement of a Gs-like G protein in TRAP secretion from the osteoclast, possibly through a cyclic AMP-dependent mechanism. PMID- 8427593 TI - Release of extracellular matrix components by bovine bone endothelial cells in continuous culture. AB - Production of collagen type I (pIp), type III (pIIIp), type IV (CIV), type VI (CVI), fibronectin (FN), laminin and glycosaminoglycans (GAGs) has been investigated in a clonal line of Bovine Bone Endothelial (BBE) cells continuously growing in culture. BBE cells produced pIp, pIIIp and FN, whereas they did not synthetize significant amounts of CIV, CVI and laminin. pIp, pIIIp and FN syntheses were influenced by cell density:pIp secretory activity reached its maximum just after cell seeding, whereas pIIIp and FN production increased later, when cells were subconfluent. Furthermore, BBE cells produced and released significant amounts of GAGs into medium. PMID- 8427594 TI - Inhibition of neuritogenic ganglioside biosynthesis in chick brain neurons by calcium ionophore A23187. AB - Treatment with Ca++ ionophore A23187 blocks biosynthesis of the major terminally (IV3) sialylated higher species of neuronal gangliosides: GD1A, GT1B, GQ1B, and Gp, key components in the plasma membrane of neurite-bearing neurons. At the same time, the neurital field diminishes markedly, resulting in neuronal reversion to pre-neuritogenic morphology. A23187 causes no biosynthetic loss of any of the simpler gangliosides that constitute the plasma membrane of pre-neuritogenic neurons, nor of ganglioside precursors leading to formation of the blocked gangliosides. The data indicate that A23187 significantly reduces neuritogenic ganglioside biosynthesis. PMID- 8427595 TI - Effect of 1,25-dihydroxyvitamin D3 on the proliferation of osteoblastic MC3T3-E1 cells by modulating the release of local regulators from monocytes. AB - Some evidence suggests an important role of mononuclear cells at the bone remodeling sites in the coupling of bone resorption to bone formation. Therefore, we examined effects of human monocytes-conditioned medium (CM) treated with 1,25 dihydroxyvitamin D3 [1.25(OH)2D3] on the proliferation of osteoblastic MC3T3-E1 cells. 1,25(OH)2D3 (10(-11)-10(-8) directly inhibited [3H]thymidine incorporation (TdR) into MC3T3-E1 cells in a dose-related fashion. 24,25(OH)2D3 and 25(OH)D3 (10(-8) and 10(-7) M) had only modest inhibitory effect, compared to that of 1,25(OH)2D3. CM, per se, on the other hand, significantly stimulated TdR, whereas CM obtained from monocytes treated with 1,25(OH)2D3(10(-10) and 10(-8) M) significantly inhibited TdR. Treatment of monocytes with 10(-7) M 25(OH)D3 significantly inhibited CM-induced stimulation of TdR to a lesser degree, compared to that of 10(-8) M 1,25(OH)2D3 and treatment of monocytes with 10(-7) M 24,25(OH)2D3 did not affect CM-induced stimulation of TdR. The inhibition of TdR by 1,25(OH)2D3-treated CM was significantly blocked by 10(-6) M indomethacin, an inhibitor of prostaglandin synthesis. Present data first indicate that 1,25(OH)2D3 inhibited osteoblast proliferation not only directly but also indirectly through modulating the release of local factors as to bone remodeling from monocytes, and also suggest that its indirect effect via monocytes was mediated at least in part through prostaglandin synthesis. PMID- 8427596 TI - Expression of biologically active human factor VIII using a baculovirus vector. AB - Factor VIII is a complex, plasma glycoprotein involved in the process of blood coagulation. Production of the recombinant molecule has largely been confined to mammalian cell systems which have, in general, proven to be inefficient producers of factor VIII. The use of a baculovirus expression system may provide increased levels of this glycoprotein, although it is not certain that insect cell-derived factor VIII will be biologically active. The N-linked glycosylation patterns in insect cells, until recently thought to be less complex than in mammalian cells, may influence activity and/or secretory ability. To this end we engineered a B domain-deleted factor VIII cDNA sequence for expression in Spodoptera frugiperda cells. The construct retained the native signal sequence to allow secretion of recombinant protein into the culture medium. Initial studies revealed the production of secreted factor VIII, and this protein was shown to possess coagulation activity. The presence of N-linked oligosaccharide residues was demonstrated, the glycosylated molecule being of a similar size to that expressed in mammalian cells. PMID- 8427597 TI - Ciliary neurotrophic factor regulates fibrinogen gene expression in hepatocytes by binding to the interleukin-6 receptor. AB - Treatment of primary rat hepatocytes with ciliary neurotrophic factor (CNTF) resulted in increased fibrinogen mRNA levels and protein expression in a time- and dose- dependent manner. The stimulation was similar to but not as potent as the response observed with interleukin-6 (IL-6). Equilibrium binding studies using radioiodinated CNTF revealed approximately 1,900 binding sites per cell with a binding affinity of 1.6-2.0 nM, indicating the presence of specific CNTF receptors on the hepatocyte surface. Binding of CNTF to the hepatocyte surface could be competed by IL-6. Additionally, IL-6 binding was also competed by CNTF, although not entirely. These findings suggest that the CNTF-induced stimulation of fibrinogen gene expression occurs, at least in part, by this cytokine's interaction with the IL-6 receptor. PMID- 8427598 TI - High affinity binding of a potassium channel agonist to intact rat insulinoma cells. AB - The specific binding of a novel tritiated K+ channel opener, [3H]BAY X 9228, has been characterized in a rat insulinoma (RINm5F) cell line. The KD was 2.1 nM and Bmax 50 fmol/mg total protein as determined by saturation analysis. The high affinity binding to intact cells was inhibited by pinacidil and by a series of BAY X 9228 analogs with an activity sequence correlating well with that for producing glyburide-reversible relaxation of partially depolarized rat aorta. This represents the first report of the specific binding of a K+ channel opener to cultured cells. PMID- 8427599 TI - Inhibition of CMP-sialic acid transport in human liver and colorectal cancer cell lines by a sialic acid nucleoside conjugate (KI-8110). AB - The sialic acid nucleoside conjugate KI-8110 has been shown to inhibit the formation of hepatic metastases from human colorectal cancer cell lines in a nude mouse intrasplenic injection model. The compound does not inhibit sialyltransferases from either human colorectal tumor cells or human liver. Transport of CMP-sialic acid into endoplasmic reticulum and Golgi vesicles is inhibited and can account for the reduction in surface sialic acid found on treated cell lines. Only a 50% inhibition of CMP-sialic acid transport could be achieved suggesting the presence of more than one transport protein with differing specificities. PMID- 8427600 TI - Phospholipase C: a putative mechanotransducer for endothelial cell response to acute hemodynamic changes. AB - Endothelial cells (EC) in vivo are exposed to a multitude of physical forces with each pulse of the cardiac cycle. Ongoing studies support the concept that EC respond to these forces through specific signal transduction pathways. Previous investigations in our laboratory have shown that EC respond to the initiation of cyclic strain or to an acute increase in cyclic strain frequency with the production of inositol 1,4,5-trisphosphate (IP3). This study demonstrates that EC also respond to an acute decrease in cyclic stretch frequency with a transient increase in IP3 production. Thus, EC detect both increases and decreases in cyclic stretch frequency with phospholipase C (PLC) activation leading to IP3 generation. PMID- 8427601 TI - Altered membrane-dystrophin association in the cardiomyopathic hamster heart muscle. AB - In the ventricular muscle from 30- to 40-day-old Bio 14.6 cardiomyopathic hamsters, the concentration of dystrophin was about 50% less than the normal controls. Despite its reduced content, extraction of dystrophin from the myopathic muscle into the microsomal fraction was threefold higher as compared to the control. These results suggest that abnormality exists in the association of dystrophin with sarcolemmal membranes in the myopathic muscle. No obvious difference, however, was observed between normal and myopathic myocytes in their immunofluorescence staining patterns. PMID- 8427602 TI - Free fatty acid-induced alterations in the steroid-binding properties of rat androgen-binding protein. AB - The free fatty acid (FFA) concentration in the epididymal cytosol of the adult rat was found to be 20-fold higher than in the serum. The binding of [3H] dihydrotestosterone to epididymal rat androgen binding protein (rABP) was modified by physiological concentrations of saturated and unsaturated fatty acids. Polyunsaturated fatty acids inhibited the binding more efficiently than monounsaturated or saturated fatty acids. Scatchard analysis and Dixon plots indicated that the number of binding sites decreased in presence of unsaturated fatty acids with an inhibition constant (Ki) of 4 microM for arachidonic acid (C20:4) and 20 microM for oleic acid (C18:1). These results indicate that unsaturated fatty acids induce alterations in rABP steroid-binding properties that could modulate the endocrine function of rABP. PMID- 8427603 TI - Stage-specific remodeling of the mammary gland basement membrane during lactogenic development. AB - Several in vitro studies have highlighted the importance of extracellular matrix proteoglycans as regulators of mammary epithelial cell differentiation. In this study monospecific antibodies to heparan (HS) and chondroitin/dermatan sulfate (CS/DS) proteoglycans were used to determine their possible significance in vivo. Immunohistochemical studies of proteoglycan distribution in virgin, pregnant, and lactating tissue demonstrated that heparan sulfate proteoglycan was found in the epithelial cell basement membrane at all stages of development. This was also true for CS/DS proteoglycans, but the relative proportions of CS and DS were strikingly modulated during lactogenesis. Notably DS glycosaminoglycans were the dominant components of the basement membrane in lactating tissue but CS glycosaminoglycans were the major proteoglycans of pregnant tissue. The possible significance of these results for gene expression in the gland is discussed. PMID- 8427604 TI - Overlapping binding sites of two different transcription factors in the promoter of the human gene for the Alzheimer amyloid precursor protein. AB - A four- to fivefold overexpression of the gene for the Alzheimer amyloid precursor protein (APP) in individuals with Down's syndrome (DS) appears to be responsible for the fifty year earlier onset of Alzheimer's disease pathology in DS compared to the normal population. Elucidation of the mechanisms regulating the expression of the human APP gene might therefore be an important step forward in understanding the processes leading to Alzheimer's disease. Recently, an activating DNA fragment proximal to the transcriptional start site of the APP gene was identified, composed of two GC-elements, A and C, both required for full transcriptional activation (1). Here I present evidence that the transcription factor Sp1 can bind to element A and that another specific complex, called C2A, can be observed, which covers a region overlapping with the Sp1 binding site on the APP promoter. Gene transfer experiments with a truncated version of the APP promoter containing mutated binding sites for the factors mentioned above support that at least two different and independent regulatory pathways for APP gene expression might exist. An imbalance between these pathways is proposed to be a putative risk factor for the development of Alzheimer's disease. PMID- 8427605 TI - Bretylium differentiates between distinct signal transducing pathways in human lymphocytes. AB - The selection of signal transducing pathways of T cells depends on the type of triggers. Antigens, antibodies or lectins induce the T cell receptor-CD3 operated pathway, and IL-2 transmits its activation signal via the IL-2 receptor. It has been demonstrated that bretylium, a quaternary ammonium ion, can significantly inhibit the first pathway at the same dose range that stimulates cell activation through the IL-2 receptor system. In the light of the different complexity of the two pathways at the plasma membrane level, and the non-toxic and reversible behavior of the drug, it is suggested that the bretylium induced sustained membrane hyperpolarization is responsible for the observation. This finding may open new possibilities in studying the mechanism of different signal transducing pathways. PMID- 8427606 TI - The Ca(2+)-sensitizing component of smooth muscle thin filaments: properties of regulatory factors that interact with caldesmon. AB - We have investigated the nature of the native Ca(2+)-sensitizing factor of smooth muscle native thin filaments. Two different types of preparation yielded protein mixtures which consistently conferred Ca(2+)-sensitivity on thin filaments reconstituted from aorta actin, tropomyosin and caldesmon. These new preparations produced a good model of native thin filaments, unlike previous systems reconstituted with calmodulin or S. 100. We conclude that the Ca(2+)-sensitizing factor is a protein of molecular weight in the range 15 kDa - 40 kDa which is heat-labile and which has properties distinct from calponin, S. 100 or calmodulin. PMID- 8427608 TI - Thomas J. O'Toole. 1993 president. PMID- 8427607 TI - Identification of the human ltk gene product in placenta and hematopoietic cell lines. AB - Two different monoclonal antibodies (MAbs) were raised against an extracellular domain and a C-terminal portion of the human ltk protein which is a receptor-type protein tyrosine kinase. Western blot analysis showed that these MAbs specifically immunoprecipitated a 100 kDa ltk protein which was transiently expressed in COS-1 cells transfected with a human ltk cDNA. By an in vitro immune complex kinase assay using these MAbs, a 100 kDa phosphoprotein was detected in human placenta and hematopoietic cell lines. These data indicate that the ltk gene product expressed in human placenta and hematopoietic cells shows tyrosine kinase activity. This is the first detection of native ltk protein naturally expressed in human cells. PMID- 8427609 TI - Research: a natural bridge. PMID- 8427610 TI - Adult-onset disorders. PMID- 8427611 TI - Missed cue. PMID- 8427612 TI - Response to John Hancock. PMID- 8427613 TI - Aural rehabilitation. PMID- 8427614 TI - Translating data into practice. PMID- 8427615 TI - The fractionation of our discipline. A time for action. PMID- 8427616 TI - Nursing home reform. The resident assessment instrument. PMID- 8427617 TI - The role of the speech-language pathologist and teacher of singing in remediation of singers with voice disorders. PMID- 8427618 TI - Caseloads of speech-language pathologists. PMID- 8427619 TI - Liability issues for occupational health nurses: an overview. AB - Occupational health nurses need to be familiar with the Nurse Practice Act within their states. Most malpractice suits against nurses are based on negligence. The occupational health nurse should also be aware of potential liability for intentional torts, such as assault and battery, breach of patient confidentiality, and informed consent. The elements of a malpractice case are: a) the duty to conform to standard of care (which is usually established by expert testimony); b) breach of that duty, or the failure to conform to the standard of care; c) a causal connection between the breach of duty and the resulting injuries; and d) actual damages suffered by the patient-plaintiff. PMID- 8427620 TI - How pesticides are handled in a rural North Carolina county: a survey of farmers. AB - Although farmers receive little guidance and training related to safety, most of the farmers in this study were knowledgeable about handling pesticides safely. The most frequently reported concern among these farmers related to the environment and health. Significant differences were noted in age, storage of pesticides, and rinsing of empty pesticide containers among certified and noncertified farmers. Recognizing other significant differences in handling practices between the certified and noncertified farmers was difficult due to the small sample size and few numbers of noncertified farmers. Although statistically nonsignificant, there were instances where the noncertified farmers and the least educated farmers responded that they always practiced particular behaviors safely. PMID- 8427621 TI - Lead revisited: a case study on lead exposed painters. AB - 1. Lead exposure, one of the oldest occupational hazards, continues to be a problem. 2. The construction industries account for the highest proportion of occupational exposures. 3. This case study found that removal of leaded paint by sanding may expose painters to lead levels above the PEL. 4. The informed occupational health nurse who "revisits" lead will be a valued team member in the identification and control of lead exposure. PMID- 8427622 TI - Occupational health nursing in the 1990s: a different model of practice. AB - 1. The American workplace is experiencing rapid change in the 1990s. Throughout the past 100 years, occupational health nursing has evolved to meet the demands of the workplace. 2. Nursing practices at some worksites are not keeping up with rapid changes impacting business and workers today. 3. A different model of occupational health nursing practice is needed to meet current and future health requirements within organizations. 4. Occupational health nurses need to identify the most critical health issues impacting their organizations and to implement a model of occupational health nursing practice designed specifically to meet those essential organizational needs. PMID- 8427623 TI - Using the words and works of others: a commentary. PMID- 8427624 TI - Facilitating return to work. PMID- 8427625 TI - OSHA's final bloodborne pathogens standard: Part II. AB - The Bloodborne Pathogens Standard has specific requirements for employee hazard communication and training as well as management of regulated waste, contaminated laundry, and housekeeping procedures. OSHA recognizes the benefits of prophylactic use of hepatitis B vaccine and requires that it be made available, at no charge, to all employees who have occupational exposure to blood or other potentially infectious materials. The goal of the Bloodborne Pathogens Standard is to reduce the occupational risk of morbidity and mortality associated with diseases such as hepatitis B and human immunodeficiency virus (HIV). PMID- 8427626 TI - Specificity of factor Xa in the cleavage of fusion proteins. AB - The precursor protein honey bee prepromelittin has been expressed as a fusion protein in Escherichia coli joined to the C-terminus of a truncated form of the bacteriophage gene 10 protein via an engineered recognition sequence for Factor Xa. Factor Xa was found to cleave poorly at the engineered site, giving a low yield of the required prepromelittin. In contrast, cleavage on the C-terminal side of the sequence VLGR at residue 67 in the gene 10 sequence proceeded in high yield. Factor Xa may be inhibited by adjacent hydrophobic sequences on the C terminal side of a potential cleavage site. PMID- 8427627 TI - The peptidase activity of human serum butyrylcholinesterase: studies using monoclonal antibodies and characterization of the peptidase. AB - Purified human serum butyrylcholinesterase, which exhibits cholinesterase, aryl acylamidase, and peptidase activities, was cross-reacted with two different monoclonal antibodies raised against human serum butyrylcholinesterase. All three activities were immunoprecipitable at different dilutions of the two monoclonal antibodies. At the highest concentration of the antibodies used, nearly 100% of all three activities were precipitated, and could be recovered to 90-95% in the immunoprecipitate. The peptidase activity exhibited by the purified butyrylcholinesterase was further characterized using both Phe-Leu and Leu enkephalin as substrates. The pH optimum of the peptidase was in the range of 7.5 9.5 and the divalent cations Co2+, Mn2+, and Zn2+ stimulated its activity. EDTA and other metal complexing agents inhibited its activity. Thiol agents and -SH group modifiers had no effect. The serine protease inhibitors, diisopropylfluorophosphate and phenyl methyl sulfonyl fluoride, did not inhibit. When histidine residues in the enzyme were modified by diethylpyrocarbonate, the peptidase activity was not affected, but the stimulatory effect of Co2+, Mn2+, and Zn2+ disappeared, suggesting the involvement of histidine residues in metal ion binding. These general characteristics of the peptidase activity were also exhibited by a 50 kD fragment obtained by limited alpha-chymotrypsin digestion of purified butyrylcholinesterase. Under all assay conditions, the peptidase released the two amino acids, leucine and phenylalanine, from the carboxy terminus of Leu-enkephalin as verified by paper chromatography and HPLC analysis. The results suggested that the peptidase behaved like a serine, cysteine, thiol independent metallopeptidase. PMID- 8427628 TI - Liquid chromatographic analysis of amino acids: using dimethylamino azobenzenesulfonyl chloride and Hypersil ODS column to analyze the composition of apo B peptides. AB - A reliable high-performance liquid chromatography (HPLC) method is described for the separation of dimethylamino-azobenzenesulfonyl-amino acid (DABS-AA). The separation is accomplished by reversed-phase chromatography on a Hypersil ODS column (4.6 x 150 mm) with a Hewlett-Packard liquid chromatography system. In addition to the developed sample and solvent preparation procedure, this precolumn modification method using dimethylaminoazobenzene sulfonyl chloride (DABS-CL) for amino acid analysis is proved reliable and sensitive. Five apolipoprotein B-100 tryptic peptides, two of them containing cysteine, were demonstrated as examples for the general application of this method in amino acid analysis. It is a useful method for analysis of cysteine- and cystine-containing peptide and, furthermore, for determination of sulfhydryl and disulfide linkages of proteins. PMID- 8427629 TI - Pectinase Aspergillus sp. polygalacturonase: multiplicity, divergence, and structural patterns linking fungal, bacterial, and plant polygalacturonases. AB - Nine forms of Aspergillus sp. polygalacturonase were purified from a commercial preparation of pectinase Rohament P using chromatographies and chromatofocusing. Individual forms differ in isoelectric point, and at least five differ in structure; whereas molecular masses and enzymatic properties are largely identical. Four forms with free alpha-amino groups have identical start positions but internal amino acid replacements. Therefore, the multiplicity is derived from true heterogeneities and not from N-terminal truncations. Peptide analysis of the major polygalacturonase reveals large variations toward the enzyme from other Aspergillus species (72-75% residue differences, depending on species) but additional similarities with the enzyme from bacterial and plant sources (only 66 71% residue differences toward the Erwinia, tomato, and peach enzymes). Combined with previous data, these facts show polygalacturonase to exhibit extensive multiplicity and much variability, but also unexpected similarities between distantly related forms with conserved functional properties. PMID- 8427630 TI - Interaction between cystatin-derived peptides and papain. AB - The interaction between papain and synthetic peptides which tentatively mimic cystatin surfaces was investigated both enzymatically and structurally. Measurements of dissociation equilibrium constants for the interaction of papain with these peptides modified by successive deletions or substitutions demonstrated that the QVVAG segment, which is highly conserved throughout members of the cystatin superfamily, is essential for the interaction. The glycyl containing (N-terminal) fragments and PW-containing (C-terminal) fragments were found to be of lesser importance, since each could be deleted without significantly modifying the interaction. These fragments improved the stability of the interacting QVVAG region, which appeared to be substrate-like in all peptides tested, as it was cleaved at the A-G bond upon peptide-papain interaction. Replacement of the A residue at the scissile bond of the QVVAG by a blocked cysteinyl residue reduced the rate of cleavage of the susceptible bond and therefore shifted the resulting peptide from a substrate to an inhibitor. Derivatization of this substituted peptide at its N- and C-terminal ends by fluoresceinyl groups resulted in a dramatic decrease in the Ki to 0.5 microM. This improvement in the inhibitory properties of the substituted and derivatized peptides was correlated with structural changes as analyzed by molecular dynamic calculations. The results were compared to those proposed for the mechanisms of inhibition by natural inhibitors of the cystatin superfamily. PMID- 8427631 TI - Hemoglobins with multiple reactive sulphydryl groups: the reaction of dog hemoglobin with 5,5'-dithiobis (2-nitrobenzoate). AB - Dog hemoglobin has four sulphydryl groups per (tetramer) molecule located at the G18(111)alpha and F9(93)beta positions. The two sulphydryls at the G18(111)alpha positions are unreactive toward nonmercurial sulphydryl reagents, but those at the F9(93)beta positions are reactive toward these reagents. We have studied the kinetics of the reaction of dog hemoglobin with 5,5'-dithiobis (2-nitrobenzoic acid) as a function of pH. At all pH values studied, the reaction is kinetically monophasic. Quantitative analysis of the pH dependence of the apparent second order rate constant shows that two ionizable groups are linked to the reaction of the sulphydryl group. Their pKa values are 5.57 and 9.0. These values are assigned to HisHC3(146)beta and to the CysF9(93)beta sulphydryl. We find that dog carbonmonoxyhemoglobin is significantly--almost an order of magnitude--less reactive than the aquomet, azidomet, and oxy derivatives. This result may be due to a greater tendency (at acid pH) for the salt bridge between HisHC3(146)beta and AspFG1(94)beta to form in the carbonmonoxy than in the other derivatives. Formation of this salt bridge is known to hinder access to the CysF9(93)beta sulphydryl [Perutz, M. F. (1970), Nature 228, 734-739]. PMID- 8427632 TI - Amino acid sequence of myoglobin from the chiton Liolophura japonica and a phylogenetic tree for molluscan globins. AB - Myoglobin was isolated from the radular muscle of the chiton Liolophura japonica, a primitive archigastropodic mollusc. Liolophura contains three monomeric myoglobins (I, II, and III), and the complete amino acid sequence of myoglobin I has been determined. It is composed of 145 amino acid residues, and the molecular mass was calculated to be 16,070 D. The E7 distal histidine, which is replaced by valine or glutamine in several molluscan globins, is conserved in Liolophura myoglobin. The autoxidation rate at physiological conditions indicated that Liolophura oxymyoglobin is fairly stable when compared with other molluscan myoglobins. The amino acid sequence of Liolophura myoglobin shows low homology (11-21%) with molluscan dimeric myoglobins and hemoglobins, but shows higher homology (26-29%) with monomeric myoglobins from the gastropodic molluscs Aplysia, Dolabella, and Bursatella. A phylogenetic tree was constructed from 19 molluscan globin sequences. The tree separated them into two distinct clusters, a cluster for muscle myoglobins and a cluster for erythrocyte or gill hemoglobins. The myoglobin cluster is divided further into two subclusters, corresponding to monomeric and dimeric myoglobins, respectively. Liolophura myoglobin was placed on the branch of monomeric myoglobin lineage, showing that it diverged earlier from other monomeric myoglobins. The hemoglobin cluster is also divided into two subclusters. One cluster contains homodimeric, heterodimeric, tetrameric, and didomain chains of erythrocyte hemoglobins of the blood clams Anadara, Scapharca, and Barbatia. Of special interest is the other subcluster. It consists of three hemoglobin chains derived from the bacterial symbiontharboring clams Calyptogena and Lucina, in which hemoglobins are supposed to play an important role in maintaining the symbiosis with sulfide bacteria. PMID- 8427633 TI - Diethylpyrocarbonate reactivity of Klebsiella aerogenes urease: effect of pH and active site ligands on the rate of inactivation. AB - Reaction of Klebsiella aerogenes urease with diethylpyrocarbonate (DEP) led to a pseudo-first-order loss of enzyme activity by a reaction that exhibited saturation kinetics. The rate of urease inactivation by DEP decreased in the presence of active site ligands (urea, phosphate, and boric acid), consistent with the essential reactive residue being located proximal to the catalytic center. The pH dependence for the rate of inactivation indicated that the reactive residue possessed a pKa of 6.5, identical to that of a group that must be deprotonated for catalysis. Full activity was restored when the inactivated enzyme was treated with hydroxylamine, compatible with histidinyl or tyrosinyl reactivity. Spectrophotometric studies were consistent with DEP derivatization of 12 mol of histidine/mol of native enzyme. In the presence of active site ligands, however, approximately 4 mol of histidine/mol of protein were protected from reaction. Each protein molecule is known to possess two catalytic units; hence, we propose that urease possesses at least one essential histidine per catalytic unit. PMID- 8427634 TI - Bothropstoxin-I: amino acid sequence and function. AB - The complete amino acid sequence of bothropstoxin-I (BthTX-I), a myotoxin isolated from Bothrops jararacussu snake venom, is reported. The results show that BthTX-I is a Lys49 phospholipase A2 (PLA2)-like protein composed of a single polypeptide chain of 121 amino acid residues (M(r) = 13,720), containing one methionine and 14 half-cystines. Although deprived of any detectable PLA2 activity, BthTX-I reveals a high degree of sequence homology with Asp49-PLA2s and with other Lys49-myotoxins. Critical mutations--such as Leu5 for Phe5; Gln11 for X11; Asn28 for Tyr28; Leu32 for Gly32; Lys49 for Asp49; and Asp71 for Asn71- which are apparently involved with the decreasing or elimination of PLA2 activity, have been detected. The same mutations occurred in myotoxin II from Bothrops asper venom, but five extra changes--namely, Pro90 for Ser90; Gly111 for Asn111; His120 for Tyr120; Phe124 for Leu124; and Pro132 for Ala132--have been found relative to myotoxin II. PMID- 8427635 TI - Partially folded rhodanese or its N-terminal sequence can disrupt phospholipid vesicles. AB - Rhodanese (thiosulfate cyanide sulfurtransferase; E.C. 2.8.1.1) is a mitochondrial enzyme that is unprocessed after import. We describe in vitro experiments showing that partially folded rhodanese can interact with lipid bilayers. The interaction was monitored by measuring the ability of rhodanese to disrupt small unilamellar vesicles composed of phosphatidylserine and to release 6-carboxyfluorescein that was trapped in the liposomes. Partially folded rhodanese, derived by dilution of urea-unfolded enzyme, efficiently induced liposome leakage. Native rhodanese had no effect on liposome integrity. Liposome disruption progressively decreased as rhodanese was given the opportunity to refold or aggregate before introduction of the liposomes. A synthetic 23 amino acid peptide representing the N-terminal sequence of rhodanese was very efficient at disrupting the liposomes. Shorter peptides chosen from within this sequence (residues 11-23 or residues 1-17) had no effect on liposome disruption. A peptide representing the tether region that connects the domains of the enzyme was also without effect. These results are consistent with the hypothesis that the N terminal sequence of rhodanese is an uncleaved leader sequence, and can interact with membrane components that are involved in the mitochondrial uptake of this protein. PMID- 8427636 TI - Essential sulfhydryl group of malic enzyme from Escherichia coli. AB - The activity of malic enzyme from Escherichia coli was unaffected by the monovalent cations Na+ or Li+ at 10 mM. At 100 mM, Li+ or Na+ inhibited the enzyme activity by 88% and 83%, respectively. However, the enzyme activity was stimulated by 40-80-fold with 10 mM K+, Rb+, Cs+, or NH4+. Less stimulation was observed with 100 mM of these stimulating cations. The stimulatory effect was lost after the enzyme was dialyzed against Tris-Cl buffer, but was regained after incubating the dialyzed enzyme with dithiothreitol. The regenerated enzyme was inactivated by 5,5'-dithiobis(2-nitrobenzoic acid). The resulting inactive thionitrobenzoyl enzyme could be regenerated to the active thiol-enzyme by dithiothreitol or converted to the inactive thiocyanoylated enzyme by KCN. The thiocyanoylated enzyme was insensitive to K+ stimulation, which suggested the essentiality of the sulfhydryl groups of the E. coli malic enzyme. PMID- 8427637 TI - Basic trypsin-subtilisin inhibitor from marine turtle egg white: hydrodynamic and inhibitory properties. AB - A basic trypsin-subtilisin inhibitor has been isolated from the egg white of marine turtle (Caretta caretta Linn.) and purified to homogeneity by gel filtration followed by ion-exchange chromatography. It has a single polypeptide chain of 117 amino acid residues, having a molecular weight of 13,600. It lacks methionine and tryptophan. Its isoelectric point is at pH 10.0 and the sedimentation coefficient (S20,w) value of 1.62 S is independent of protein concentration. It has a Stokes radius of 18.8 A, an intrinsic viscosity of 0.048 dl g-l and a diffusion coefficient of 10.17 x 10(-7) cm2 sec-1. Its fluorescence emission spectrum is similar to that of free tyrosine and the bimolecular quencing rate constant of its tyrosine residues with acrylamide is 3.15 x 10(9) M 1 sec-1. The inhibitor strongly inhibits both trypsin and subtilisin by forming enzyme-inhibitor complexes at a molar ratio of unity. The nature of inhibition toward both enzymes is not temporary. It has independent binding sites for inhibition of trypsin and subtilisin. Chemical modification with tetranitromethane suggests the presence of three tyrosine residues on the surface of the inhibitor molecule. PMID- 8427638 TI - The complete amino acid sequence of ribosomal protein S8 from Thermus thermophilus. AB - Protein S8 from Thermus thermophilus consists of 138 amino acids of M(r) 15,840. Its primary structure was established using peptide sequences from two different digests. Protein S8 from T. thermophilus shares a high percentage of identity with protein S8 from Thermus aquaticus. There are some consensus sequences between proteins S8 from eubacteria, archebacteria, chloroplasts, and cyanelles. PMID- 8427639 TI - Primary structure of rabbit lens alpha-crystallins. AB - The primary structure and posttranslational modifications of rabbit lens alpha crystallins were examined using electrospray ionization mass spectrometry to determine the molecular weights of the intact proteins and fast atom bombardment mass spectrometry to analyze proteolytic digests of the alpha A- and alpha B crystallins. The previously determined primary structure of alpha A-crystallin was confirmed. Posttranslational modifications detected included one phosphorylation site and the presence of a truncated form minus the five C terminal residues. The previously undetermined amino acid sequence of rabbit alpha B-crystallin was determined to be the same as the bovine alpha B-crystallin sequence except at three residues: Thr 40, Thr 132, and Pro 153. Rabbit alpha B crystallin showed evidence of phosphorylation at the same three sites as bovine alpha B-crystallin. The molecular weights of the intact proteins indicated that any one molecule had a maximum of two phosphorylations. Also, there was a truncated form which did not include the five C-terminal residues. PMID- 8427640 TI - Full-thickness pressure ulcers: patient and wound healing characteristics. AB - To investigate the patient and healing characteristics related to full-thickness pressure ulcers, 119 consecutive patients admitted with ulcers in three acute care, four longterm care, and one rehabilitation agency were studied. Of the 119 patients with 153 pressure ulcers, only 48 (40%) had full-thickness ulcers. Compared to patients with partial-thickness ulcers, patients with full-thickness ulcers were more likely to have multiple ulcers, occasional incontinence of urine and feces, a compromised overall skin condition, and a less than optimal nutritional status at baseline. Full-thickness ulcers treated with a hydrocolloid dressing (DuoDERM Hydroactive) did not develop adverse reactions; clinicians perceived the dressing to be efficacious. Ulcers that healed during the study decreased 47% in area in two weeks. This distinguished ulcers that healed from those that did not heal. The findings suggest that ulcers that do not decrease in size within two weeks should be reevaluated for additional or alternate treatments. PMID- 8427641 TI - Relationship between body weight, body position, support surface, and tissue interface pressure at the sacrum. AB - This descriptive, comparative study examined the relationship between three body positions, body weight, and tissue interface pressure measurement on three support surfaces among residents who were 65 years of age or older. The support surfaces included the Sof. Care bed cushion, Biogard foam mattress, and hospital mattress. Subjects included 12 ideal body-weight and six less-than-ideal body weight individuals from a long-term care facility in a Mid-western state. Results indicated no significant difference in sacral tissue interface difference between ideal body weight and less-than-ideal body weight subjects. However, the sacral measurements were significantly greater in the supine position than those taken in the Fowler's and semi-Fowler's positions. A significant interaction was found between position and support surface with subjects in Fowler and semi-Fowler positions on the Sof. Care Surface exhibiting the lowest mean tissue interface pressure at the sacrum. PMID- 8427642 TI - Seeking quality care for patients with pressure ulcers. AB - Improving quality care for patients with pressure ulcers resulted in a two-year intensive effort to educate staff, keep morale high, standardize care plans by stage of ulcer, and participate in a comparison study of two support surfaces (Thera-Pulse, Kinetic Concepts and Geo-Matt, SpanAmerica). When all pressure ulcers were considered, the analysis of covariance revealed no statistically significant difference in the healing of pressure ulcers with respect to type of support surface used (F[1, 78] = 0.35, p > .05). For patients with stage III and IV pressure ulcers, the proportion of patients improving more than 10cm2 was higher in the air-suspension group. There was relatively little difference in the Stage II ulcer patients. PMID- 8427644 TI - [The 50th anniversary of the Hospital Infantil de Mexico Federico Gomez]. PMID- 8427643 TI - Reassessment of the use of genuine sheepskin for pressure ulcer prevention and treatment. AB - The use of genuine sheepskin was compared to the use of synthetic sheepskin in two long term care institutions. Data for the control group of 44 residents placed on synthetic sheepskin were obtained by retrospective chart audit. Sixty four residents were randomly placed on genuine merino sheepskin (WoolTec USA, Maspeth, NY). The two groups were similar in age, diagnosis, polypharmacy, and risk assessment score. In the control group 41% maintained skin integrity, 38% improved, and 21% deteriorated; in the study group 63% maintained skin integrity, 37% improved, and none deteriorated. The use of genuine sheepskin is recommended to reduce the risk factors of friction and shear. PMID- 8427645 TI - [The prevalence of antibodies against Giardia lamblia in umbilical cord serum and in maternal peripheral blood]. AB - Using an immunoenzymatic assay (ELISA), 40 serum samples from the peripheral blood of women in labor and serum from the umbilical cord of their offspring were analyzed in order to detect the prevalence of antibodies against Giardia lamblia. The results show that 12.5% of the maternal serum samples and 15.0% of those from the umbilical cord had optical density values comparable to those positive for G. lamblia used in the assay. This suggests that important concentrations of maternal IgG antibodies for the parasite are transferred through the placenta. The role that these antibodies play in the neonatal protection is still to be determined. PMID- 8427646 TI - [Recognition of the reviewers of the Boletin Medico del Hospital Infantil de Mexico]. PMID- 8427647 TI - [Residence close to high-tension electric power lines and its association with leukemia in children]. AB - INTRODUCTION: There are different risk factors which have been related to the presence of leukemia in children. In the past years one of these factors has become relevant, the risk of living in an area near to high electric voltage lines, generators of electromagnetic fields of low frequency (EMF), which can cause development of leukemia in children. OBJECTIVE: To learn whether living in an area close to EMF generator sources, electric transformers, high electric voltage distribution or transmission lines and electric substations, is a risk factor in the development of leukemia in children living in Mexico City. MATERIAL AND METHODS: A projective study of case-control was accomplished. The cases were obtained from hospitals of the third level. The diagnosis of leukemia in its different varieties was confirmed through biopsy of bone marrow. The controls were selected in the same hospital from inpatients and outpatients with different problems, except neoplasia. A total sample of 81 cases and 77 controls was analyzed. The residence of the controls and cases were visited using a questionnaire coded with the different study variables. To diminish the memory bias in relation to EMF, the subjects were shown different pictures pointing out the different sources of exposures, which were asked. Having obtained the information, different odds ratios (OR) were calculated for the different associations, as well as the confidence intervals at 95% and an unconditioned logistic regression was accomplished to know the adjusted OR. RESULTS: There were no differences between the cases and controls according to the relative who gave the information, the current age of children, the parents's age, the social class and the parent's occupation. It was found that all the generating sources of EMF, which were involved in the study, had and OR above 1. Being the highest, the ones living near the distribution or transmission wires of high voltage with an OR of 2.63 (1.26-5.36) and 2.5 (0.97-6.67) respectively. When the distance of exposure was controlled, the highest OR was for distribution lines (OR 2.12; 0.79-5.85). When the analysis was applied to persons who have moved from the residence, it was found that the OR was above 1 in all the associations, the highest being the distribution wires and with the distance the highest was the electric substations. Furthermore, the multivariate analysis showed that the risk continued only for the distribution wires. CONCLUSIONS: The EMF exposure was found positive, however this is not very precise, that is why it is necessary to carry out other studies to confirm the existence of the association and correct possible biases which could appear during the research. PMID- 8427648 TI - [Laparoscopic surgery. Experience with the first pediatric surgical patients]. AB - The surgery technique and technology are changing. Nowadays, the laparoscopic surgery, endoscopic surgery or minimum invasive surgery are done routinely at many hospitals. General surgeons, gynecologists, thoracic surgeons and oncologists are using this kind of surgery frequently. The laparoscopic surgery, done by trained surgeons, no matter their specialty, has demonstrated many advantages over the traditional surgery: less post operation painful, less chirurgical time and hospitalization; better esthetics results; less complications; early ambulation and back to normal life in very short time. Today in Mexico there are no reports concerning pediatric patients, operated by pediatric surgeons using laparoscopic surgery. Our purpose is to show our experience in laparoscopic surgery and the results of the first five children who have been operated by this method. PMID- 8427649 TI - [The indigestion cure: a common and dangerous practice]. AB - PURPOSE: To identify current concepts and practices to treat a folk illness "empacho" in Leon, Guanajuato and a critical appraisal of risk involved. DESIGN: Prospective cross sectional survey. FRAMEWORK: General population. PATIENTS: A random sample of 519 families. MEASUREMENTS: Socioeconomic stratum, the point of view on empacho, the type and kind of treatment undertook and the clinical course observed were recorded. RESULTS: The 45.9% of the families considered empacho as a serious condition, though the prevalence of the concept was less in the high socioeconomic group. The most frequent (44.1%) concept on empacho was "something stuck in the gut" and it use to be deal with abdominal massage and herb infusions. Cooking oil was used in 34.7%, bismuth powder in 36.5%, and an unknown powder in a further 7.9% of cases. The remedy was "prescribed" by the mother herself in 48.3% of patients, whilst in the remaining someone else did it. Simultaneous medical treatment was received by 44.2% of children and 95% of them improved. CONCLUSIONS: There is a high prevalence of the concept of empacho in the general population, mainly in the low and medium socioeconomic strata. In its treatment, dangerous practices are used, which can results in lipidic pneumonia or poisoning and according to their intensity can produce lose of life or deterioration in the respiratory or neurologic functions or both. PMID- 8427650 TI - [Angelman's syndrome (happy puppet) in 2 siblings. A follow-up over 10 years]. AB - The Angelman syndrome or "happy puppet" syndrome is a disorder of severe mental retardation, seizure, paroxysms of laughter, absent speech, jerky movements and ataxic gait. We present two sibs, man and woman, with this disorder, fact that support the possible autosomal recessive inheritance as a cause of this pathology, which hereditary mechanism is still a controversial point. Besides, we can observe different expression, being the woman more severely affected than the man. To our knowledge, this is the first mexican family reported with this syndrome, and with a ten years follow up. Chromosomal studies, with high resolution technique, were normal, we did not find the 15 chromosomic deletion referred as a possible cause in some cases, that is why it is undeniable that genetic heterogeneity exists in this syndrome. PMID- 8427651 TI - [A giant follicular cyst of the ovary in a newborn infant]. AB - The tumoral lesions proceeding of ovary in the newborns are the 3-6% of all abdominal tumors. The most frequent tumor is ovarian follicular cyst which will be diagnosed prenatally. As a result of ovarian stimulation by fetal gonadotrophins, maternal estrogen and placenta + gonadotrophins. Maternal antecedents reported diabetes, toxemia and isoimmunization. The most frequent complication is torsion. We present the case of one female newborn, with abdominal mass present by abdominal palpation referred of the first gestation of mother with 27 years of age, with primary sterility of 11 years without treatment, with toxemia during the pregnancy. The abdominal x-film show a intraperitoneal mass the ultrasound show cystic and solid mass, without determined origin, and the scan magnetic resonance (MR) a tumoral mass with origin in the pelvic hole with extension to the infrahepatic zone. The surgical finding was left ovarian cyst with left oophorectomy and salpingectomy. PMID- 8427652 TI - [The verbal autopsy: a tool for the study of mortality in children]. AB - The verbal autopsy (VA) is a technique that has been used since 1931, but it wasn't until the last decade that it has been more widely used by different investigators to study mortality both in infancy/childhood and adulthood. The VA consists of an interview directed to a care-giver (usually the mother) close to the deceased subject, and its objective is to disclose information about the cause of death. The VA has been particularly useful in those places where a reliable record of mortality is unavailable or nonexisting. This paper describes the assumptions on which the VA is based, and highlights its most important methodological aspects. Lastly, we stress the use of the VA as an useful too to assess the process associated with mortality, including the family's decision making process to look for medical care as well as the response and characteristics of the medical care system. In this respect, we point out to the usefulness of the VA as an interface between epidemiology and ethnography, and stress its potential impact as a tool to learn more about the process of health seeking behaviors, from the points of view of mothers and physicians. PMID- 8427653 TI - [Diabetic ketoacidosis]. AB - Is the most commun and serious complication of the diabetes mellitus in the childhood and adolescent, the current mortality rate in most tertiary hospitals for children is about 2 percent. Is the metabolic consequence of a deficiency of insulin, it can be precipitating for stress, or inappropriate knowledge of the management and in some cases of subclinical diabetes or it still has not been diagnostic. The majority of instances of diabetic ketoacidosis can be prevented by early recognition of ketosis, and on appropriate ambulatory management. PMID- 8427654 TI - [The verbal autopsy on children with a respiratory infection and acute diarrhea. An analysis of the disease-care-death process]. AB - The study focuses on children between 72 hours and five years of age who died of acute respiratory infection (ARI) or acute diarrhea (AD) in the State of Tlaxcala. Peer Review Mortality Committee of the State contributed with the staff to the deaths analysis. Cases were included only when diagnosis was confirmed by verbal autopsy (VA). One hundred and thirty two cases were included (98 corresponding to ARI deaths and 34 to AD). The process related to medical care seeking behaviors and prescribing practices by private and non-private physicians was analyzed through the VA. During the study period, 60% of children with ARI and 58.9% of children with AD died at home. More than 80% of these children had received medical care within three days preceding their death, and 50% of them had been seen by a physician within 12 hours prior to their death. Most of these visits were to a private doctor (71% for ARI and 86% for AD). Forty seven percent of treatments prescribed for ARI were judged to be wrong, either because of a bad choice of antibiotic or because the physician did not prescribe an antibiotic when the patient required it. Similarly, 65% of treatments for AD were considered erroneous, either due to the use of an antibiotic which was not justified or due to the lack of oral rehydration therapy when it was needed. Additionally, late referral to a hospital was considered as having direct influence at the death in half of the consultation. Families were too late in demanding medical care or demanded no care at all in 21.9% of cases of ARI and in 6.1% of cases of AD. We have found the VA to be useful in identifying problems related to the process of health-seeking behaviors and medical care. Our results suggest interventions that may lower the high mortality rates in Tlaxcala, such as training workshops directed to institutional and private physicians, and the implementation of top of-line treatment centers where high-risk patients can be referred and also the health care workers can learn the correct treatment of both diseases. Future studies should focus on the identification of alarm signs and risk factors that may help to lower mortality due to ARI or AD, when recognized and treated at early stages. PMID- 8427655 TI - [Bacterial meningitis, the experience at the Instituto Nacional de Pediatria, 1986-1988]. PMID- 8427656 TI - Ambulatory blood pressure monitoring in normotensive and hypertensive type 2 diabetes. Prevalence of impaired diurnal blood pressure patterns. AB - To assess the prevalence of an impaired diurnal blood pressure (BP) pattern in a population of both normotensive and hypertensive diabetics, noninvasive ambulatory BP monitoring (SpaceLabs 5200, Redmond, WA) was performed in 96 outpatients with type 2 diabetes (47 normotensives and 48 hypertensives) and in 103 control subjects without diabetes (50 normotensives and 53 hypertensives). Mean 24 h and daytime (06:00 to 22:00) BP and heart rate (HR) were not statistically different in diabetic patients compared to nondiabetic ones. Nighttime (22:00 to 06:00) BP and HR tended to be higher in both normotensive and hypertensive diabetics, although not significantly. Heart rate, diastolic BP (DBP), and especially the nocturnal systolic BP (SBP) decrease, were less marked in both normotensive and hypertensive diabetics, with a consequent increase in rate-pressure. A significant correlation was found between the percent decrease in nighttime SBP and the decrease in orthostatic SBP in casual BP measurements. The analysis of individual recordings allowed us to detect an impaired circadian pattern (the disappearance of the nocturnal BP decrease or a paradoxical BP increase) in 30% of the normotensive and 31% of the hypertensive diabetics (v 6% of the normotensive and 6.4% of the hypertensive nondiabetic subjects). As the absence of a nocturnal BP fall has been associated with the increased prevalence of left ventricular hypertrophy and atherosclerotic cardiovascular disease, its detection by ambulatory monitoring might be of prognostic and therapeutic importance. PMID- 8427657 TI - Endogenous erythropoietin and salt sensitivity of blood pressure in patients with essential hypertension. AB - To investigate a possible involvement of endogenous erythropoietin (EPO) in the salt sensitivity of blood pressure in essential hypertensive (EHT) patients, plasma EPO concentrations were measured during different salt intakes in 14 patients with EHT. All patients were given low salt (34 mmol NaCl/day) and high salt (342 mmol NaCl/day) diet of 7 days each. The plasma EPO concentrations were significantly higher on the high salt diet than those of low salt diet (23.5 +/- 1.9 v 18.7 +/- 1.8 mIU/ml, mean +/- SD, P < .05). The percentage change of plasma EPO concentration with salt loading correlated positively with hematocrit (Ht) at high salt diet (r = -0.62, P < .02) and tended to be correlated with plasma hemoglobin at high salt diet (r = 0.52, P < .10). These results suggest that the secretion of EPO is increased in response to hemodilution caused by the salt loading and the increased EPO concentration in plasma which may contribute to the increase in blood pressure through an expansion of total blood volume due to an enhanced red cell generation in combination with salt and water retentions. PMID- 8427658 TI - Hypotensive effect of losartan, a nonpeptide angiotensin II receptor antagonist, in essential hypertension. AB - We examined the chronic effects of losartan (DuP 753), a novel orally active angiotensin II receptor antagonist, on blood pressure and renal function in eight hospitalized patients with essential hypertension. After a control period of 1 week, losartan was administered orally once a day for 2 to 4 weeks in increasing doses of 12.5, 25, 50, and 100 mg, until blood pressure in the supine position decreased more than 20 mm Hg (systolic) and 10 mm Hg (diastolic) (or 13 mm Hg in mean blood pressure). The average dose of losartan was 59.4 +/- 43.7 (mean +/- SD) mg/day. Systolic, diastolic, and mean blood pressures, according to 24 h monitoring, fell significantly, from 151.9 +/- 6.8 to 137.2 +/- 7.9 mm Hg, from 90.6 +/- 3.7 to 81.0 +/- 3.7 mm Hg, and from 111.1 +/- 4.6 to 99.7 +/- 5.0 mm Hg, respectively (mean +/- SE, P < .01 for each), with no change in circadian rhythm or variability of blood pressure. Reduction in blood pressure was slightly greater during daytime than during sleep time. Unlike peptide angiotensin II antagonists, losartan did not exert pressor action. No significant alterations were observed in body weight, serum electrolytes, creatinine clearance, urine volume, or urinary excretion of sodium. Losartan significantly lowered serum uric acid concentration from 5.5 +/- 0.4 to 4.8 +/- 0.3 mg/dL (P < .05). Urinary excretion of uric acid increased significantly from 498.9 +/- 64.4 to 540.6 +/- 66.6 mg/day (P < .05). Plasma renin activity rose significantly but plasma aldosterone concentration did not change with the losartan treatment. These results suggest that losartan has a long-acting hypotensive effect with a hypouricemic action in essential hypertension. PMID- 8427659 TI - Defective 3,4-dihydroxyphenylalanine decarboxylation to dopamine in hydralazine treated hypertensive patients may be pyridoxine remediable. AB - The previously observed defective dopamine (DA) generation from 3,4 dihydroxyphenylalanine (DOPA) can also be seen in patients treated for many years by hydralazine. This may be due to a hydralazine-induced depletion of pyridoxine, an essential coenzyme of the aromatic L-amino acid decarboxylase (LAAD). Eleven hydralazine-treated stable essential hypertensive (EH) patients, initially found to have a defect in the DOPA decarboxylation to DA, tested by a single DOPA administration (500 mg, orally), were retested by the same test 4 days after pyridoxine pretreatment (100 mg/day) for data on blood pressure (BP), pulse rate, and renal and plasma catecholamines and their metabolites, as well as plasma atrial natriuretic factor (ANF), cyclic GMP (cGMP), plasma renin activity (PRA), and plasma aldosterone (PA). Initially, hydralazine-treated stable EH patients manifested, following DOPA administration, lower DOPA decarboxylation to DA than control subjects. Pyridoxine pretreatment accelerated DA generation from exogenous DOPA and attenuated the DOPA-induced increases in plasma and urinary DOPA and its metabolite 3-O-methyl-DOPA, but accentuated the increase in free DA and its main metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), while BP, ANF, cGMP, PRA, and PA remained unaffected. The DOPA induced increments of urinary DA were, in contrast to plasma DA changes, blunted by pyridoxine pretreatment. The attenuation of the sodium excretion by pyridoxine pretreatment exceeded that of the DA excretion, suggesting that pyridoxine suppressed a natriuretic factor, other than ANF, or activated a sodium-retaining factor, other than renin or aldosterone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427660 TI - The dose-dependent reduction in blood pressure through administration of magnesium. A double blind placebo controlled cross-over study. AB - Seventeen patients with a diastolic blood pressure over 90 mm Hg were recruited from a running health screening program to participate in a double blind cross over study of magnesium supplementation (15 mmol Mg2+/day (Emgesan, Kabi Pharmacia) for 3 weeks, followed by 30 mmol Mg2+/day for another 3 weeks, finishing with 40 mmol Mg2+/day for a final 3 weeks). A significant decrease in the mean systolic blood pressure was recorded from 154.0 +/- 10.7 mm Hg to 146.1 +/- 16.9 mm Hg (P = .031) while the mean diastolic blood pressure decreased from 100.2 +/- 4.2 mm Hg to 92.0 +/- 6.6 mm Hg (P = .0001). PMID- 8427661 TI - Racial differences in platelet cytosolic calcium and calcitropic hormones in normotensive subjects. AB - Elevated levels of serum parathyroid hormone (PTH) and platelet cytosolic free calcium ([Ca2+]i) have been reported in subjects with essential hypertension. In addition, there is a positive correlation between serum PTH and platelet [Ca2+]i in white subjects with essential hypertension. Black normotensive subjects have relatively higher levels of serum PTH when compared to white normotensive subjects. To investigate the possibility that elevated serum PTH levels in black normotensives may contribute to elevated platelet [Ca2+]i, calcitropic hormone profiles and platelet [Ca2+]i were determined in 31 black normotensive subjects and 34 age-matched white normotensive subjects. There was no difference between the two groups in total serum calcium, plasma ionized calcium, or creatinine clearance. However, serum PTH was significantly elevated (P < .02) in the black normotensive group. Serum 1,25(OH)2 vitamin D levels were similar between the two groups whereas serum 25(OH) vitamin D levels were significantly lower (P < .001) in the blacks. The 24 h urinary excretion of Ca was also lower (P < .05) in the black normotensive group. Basal platelet [Ca2+]i was significantly lower (P < .05) in black normotensive than in white normotensive subjects. Serum PTH levels did not correlate with platelet [Ca2+]i in either group, or in the groups combined. These results demonstrate that the higher serum PTH concentrations in black normotensives is not associated with higher platelet [Ca2+]i, as is the case in white hypertensive patients. PMID- 8427662 TI - The lipoxygenase inhibitor phenidone is a potent hypotensive agent in the spontaneously hypertensive rat. AB - Previous studies from our laboratory indicated that the lipoxygenase inhibitor phenidone markedly attenuates angiotensin II (AII) induced vascular contractility. Phenidone was also shown to inhibit the formation of vascular lipoxygenase products and to reduce blood pressure in the AII-dependent renovascular hypertensive rat. We have now examined the effects of phenidone in the spontaneously hypertensive rat (SHR). A single dose of phenidone lowered intraarterial systolic pressure in a dose dependent manner in both SHR and Wistar Kyoto (WKY) [(max 74 +/- 15 and 22 +/- 3 mm Hg, respectively; P < .001)], but the effect was substantially greater in SHR. Long-term oral phenidone administration arrested the evolution of hypertension in 6 week old SHR treated over a period of 4 weeks (control 190 +/- 2 mm Hg; phenidone treated rats 164 +/- 4 mm Hg; P < .01). To assess the role of AII related mechanisms in the hypotensive effect of phenidone, the acute effect was studied in SHR on high and low sodium intake. In addition, the effect of captopril was compared to that of phenidone alone or captopril and phenidone in salt restricted SHR. While a single dose of phenidone (30 mg/kg, intraperitoneally) elicited similar maximal effects in SHR on high and low sodium intake (54 +/- 6 and 52 +/- 5 mm Hg compared to basal blood pressure, respectively), the hypotensive effect in sodium restricted rats was more sustained. Phenidone had no further hypotensive effect in captopril treated, salt restricted SHR.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427663 TI - Effect of intravenous calcium infusion on indices of activity of the parathyroid glands and on urinary calcium and sodium excretion in normotensive and hypertensive subjects. AB - The effect of an intravenous calcium gluconate load (10 mg/kg over 5 min) on plasma ionized calcium concentration, parathyroid hormone (PTH), and the rate of urinary excretion of calcium, sodium, and nephrogenous cyclic adenosine monophosphate (NcAMP) was examined in 26 patients with essential hypertension and 27 age- and sex-matched normotensive subjects. Prior to calcium administration hypertensives had lower plasma ionized calcium concentration (P < .01) and higher PTH levels (P < .001) and excreted more calcium (P < .05) and NcAMP (P < .001) in the urine compared to normotensives. Following calcium infusion, plasma ionized calcium did not differ significantly between the two groups, but PTH levels remained higher in the hypertensive subjects at both 60 min (P < .001), and at 120 min (P = .02) post-load. Post-load values for both urinary calcium excretion and urinary sodium excretion were significantly higher in the hypertensive subjects than in the control group. Both before and after calcium infusion, urinary calcium excretion was positively correlated with urinary sodium excretion in each of the two groups, but for the same level of sodium excretion, hypertensives excreted more calcium in the urine, compared to normotensives, both before (P < .05) and after calcium infusion (P < .001). A positive correlation between basal plasma renin activity (PRA) values and plasma ionized calcium values obtained before (r = 0.42, P = .03) or at 60 min (r = 0.41, P = .03) and 120 min (r = 0.42, P = .03) after calcium infusion existed only in the hypertensive subject group. Post-load urinary sodium excretion values were negatively correlated to basal PRA values in both groups (r = -0.55, P < .01 and r = -0.58, P < .01 for hypertensives and normotensives, respectively), but a similar negative correlation between post-load urinary calcium excretion values and basal PRA values existed only in the hypertensive subject group (r = -0.50, P < .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427665 TI - DOCA-salt hypertensive rat hearts exhibit altered expression of G-proteins. AB - We have recently demonstrated an alteration in the levels of G-proteins and their correlation with adenylyl cyclase in spontaneously hypertensive rats (SHR). In the present studies we examined if the other models of hypertensive rats, such as DOCA-salt hypertensive rats (HR), also exhibit the similar alterations in G protein and in adenylyl cyclase activity. We have determined the adenylyl cyclase activity stimulated and inhibited by hormones, as well as the levels of G proteins using specific antibodies and cDNA probes in the hearts from DOCA-salt HR and their sham-operated controls after 2 and 4 weeks of treatment. Adenylyl cyclase activity stimulated by GTP gamma S, isoproterenol, and glucagon was significantly decreased in heart sarcolemma from DOCA-salt HR as compared to their controls after 2 and 4 weeks of treatment. In addition, the inhibitory hormones inhibited the enzyme activity to a greater extent in hypertensive rats than controls. Furthermore, the levels of Gi alpha-2 and Gi alpha-2 mRNA, as determined by immunoblotting and Northern blotting techniques, respectively, were higher in hearts from DOCA-salt HR. However, the levels of G8 alpha 45 were decreased in these rats. These results indicate that, similar to SHR, the hearts from DOCA-salt HR exhibit the increased expression of Gi, however unlike SHR, the expression of G8 was decreased. It is suggested that the altered expression of G proteins may partly be responsible for the decreased responsiveness of adenylyl cyclase to hormone stimulation and increased responsiveness to hormone inhibition in DOCA-salt hypertension. PMID- 8427664 TI - Improved accuracy of indirect blood pressure measurement in patients with obese arms. AB - We studied 20 patients with moderate to severe obesity (body mass index 37 +/- 8 kg/m2) with obese arms (arm circumference 37 +/- 4 cm). Their blood pressure was measured directly in the brachial artery in one arm and simultaneously indirectly in the other arm using either a large standard cuff (rubber bag 12 x 35 cm) or a new cuff (the Tricuff, Pressure Group AB, Stockholm) containing three rubber bags of different sizes, which automatically selected the appropriately sized bag in relation to arm circumference. Both cuffs showed a significant overestimation of the diastolic blood pressure (standard cuff 13 mm Hg, P < .001, Tricuff 6 mm Hg, P < .01). The error of the standard cuff was significantly greater than that of the Tricuff (P < .001). The differences in systolic blood pressure between the intraarterially and the indirectly measured results were small (0.2 to 3 mm Hg) and not statistically significant with either cuff. The new Tricuff offered an advantage as compared to a large standard cuff in terms of a lesser overestimation of the diastolic blood pressure. In practical terms, this should lead to a reduction in the overestimation of diastolic hypertension in obese individuals. PMID- 8427666 TI - Family history studies in hypertension research. Review of the literature. AB - Reviewed in the present article are over 150 family history studies of essential hypertension. By comparing normotensive individuals with and without a family history of hypertension, these investigations seek to identify potential pathophysiologic factors that predate the development of high blood pressure. The research literatures summarized here represent four general areas: 1) cellular salt transport mechanisms, 2) dietary sodium, intravascular volume, and renal function, 3) cardiovascular morphology and physiology, and 4) cardiovascular reactivity. There is strong evidence of early cardiac morphologic changes (greater left ventricular wall thickness and mass) and altered peripheral vascular capacity and responsivity to pressor stimuli among normotensive individuals with a positive family history. In contrast, cardiac output, sodium consumption, intravascular volume, and cardiovascular responses to isometric exercise and standing do not differ in persons with and without a family history of hypertension. Other articles are characterized by inconsistent results, which may be a reflection of the heterogeneity of essential hypertension, but also may be due to methodological weaknesses. The latter include failure to confirm the blood pressure status of ostensibly hypertensive or normotensive family members and the use of relatively weak study designs (eg, where a positive history is defined by a single, hypertensive first-degree relative). PMID- 8427667 TI - Prevalence of extracranial carotid artery lesions at duplex in primary aldosteronism. AB - Renovascular hypertension and high renin hypertension were found to be associated with an excess prevalence of carotid artery atherosclerotic lesions and to a higher risk of stroke, respectively, as compared to low-to-normal renin hypertension. Primary aldosteronism, being characterized by hypertension and a chronically suppressed plasma renin activity, should be accompanied by a low prevalence of carotid artery lesions. To verify this hypothesis we investigated prospectively, by a high resolution duplex ultrasound technique, the prevalence of extracranial carotid artery lesions in a case-controlled study of 34 (22 women and 12 men, aged 22 to 76 years) patients with no history or symptoms of cerebrovascular disease. Primary aldosteronism was diagnosed in 17 patients; 12 had a surgically confirmed unilateral aldosterone-secreting adenoma; and 5 had idiopathic hyperaldosteronism. Each primary aldosteronism patient was individually matched with a control with primary hypertension for sex, race, age, body mass index, casual blood pressure levels, duration of hypertension, smoking, diabetes mellitus, total serum cholesterol, and triglycerides. After the matching, the two groups were similar in terms of demographic features and overall cardiovascular risk profile (all P = NS). However, plasma renin activity and aldosterone levels were significantly lower and higher, respectively, in primary aldosteronism than in primary hypertensive patients. In primary aldosteronism the overall prevalence of carotid artery lesions at duplex was 59%, not significantly different from that (53%) found in primary hypertensives. Thus, at variance with renovascular hypertension, primary aldosteronism is not associated with an excess prevalence of carotid artery lesions. PMID- 8427668 TI - Commentary on Shigetomi and Kuchel's work on DOPA decarboxylation. PMID- 8427669 TI - Caffeine and ambulatory blood pressure. PMID- 8427670 TI - The value of the Hawksley Random-Zero Sphygmomanometer. PMID- 8427671 TI - Validating ambulatory blood pressure monitors. PMID- 8427672 TI - The coronal approach. PMID- 8427673 TI - The use of submentoplasty. PMID- 8427674 TI - The new certificate of added qualifications in pediatric otolaryngology. PMID- 8427675 TI - Black-box mathematics and medical practice. PMID- 8427676 TI - The relation of hearing in the elderly to the presence of cardiovascular disease and cardiovascular risk factors. AB - Hearing loss with age (presbycusis) is a substantial problem for the elderly. To investigate the possible relation of presbycusis to cardiovascular disease (CVD), the hearing status of a cohort of 1662 elderly men and women was determined and compared with their 30-year prevalence of cardiovascular disease. Age-adjusted multivariate logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (95% CIs) to describe the relation of hearing to cardiovascular disease events, cardiovascular disease risk factors, and both events and risk factors separately for the 676 men and for the 996 women. Cardiovascular disease events were the sum of coronary heart disease, stroke, and intermittent claudication. Five groups of risk factors were studied: hypertension and blood pressure; diabetes, glucose intolerance, and blood glucose level; smoking status and number of pack-years of cigarettes; relative weight; and serum lipid levels, including cholesterol, triglycerides, and lipoprotein fractions. Low-frequency hearing (low pure-tone average, 0.25 to 1.0 kHz) was related to cardiovascular disease events in both genders but more in the women. For women, the OR of having any cardiovascular disease event for a low pure-tone average of 40 dB hearing level was 3.06 (95% CI, 1.84 to 5.10); for a high pure-tone average (average of 4 to 8 kHz) of 40-dB hearing level, the OR for any cardiovascular disease event was 1.75 (95% CI, 1.28 to 2.40). In men with a low pure tone average of 40-dB hearing level, the OR for stroke was 3.46 (95% CI, 1.60 to 7.45) and for coronary heart disease the OR was 1.68 (95% CI, 1.10 to 2.57).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427677 TI - Acquired atresia of the external auditory canal. Surgical treatment and results. AB - Atresia of the external ear canal following recurrent external otitis or surgery for chronic otitis media was treated in 17 ears with a canalplasty procedure using a retroauricular incision. The stenotic meatus was widened by removing fibrous tissue, and the tympanic membrane was deepithelialized. The posterior, superior, and inferior bony canal was widened until the first mastoid cells were encountered. The anterior canal was widened in cases where there was an anterior bony overhang. The bony canal was lined with split-thickness skin grafts. Meatoplasty was performed, and split-thickness skin grafts were grafted onto the margins of the meatoplasty to cover the lateral part of the ear canal. Preoperative and postoperative surgical results, including hearing levels, are presented and compared with those from the few other series from the literature. PMID- 8427678 TI - Systemic osteoclast activation by localized pressure. AB - It has been demonstrated that positive air pressure applied to the bulla of the Mongolian gerbil results in significant osteoclastic activation and bone resorption. In this study, continuous positive air pressure of 15 mm Hg was applied to the right bulla for 1 week. A significant increase in osteoclast number and surface were noted in the contralateral, unpressurized bulla as well as in a distant site, the upper extremity (radius). Possible mechanisms are discussed by which a local stimulus (pressure) may cause distant osteoclast activation. PMID- 8427679 TI - Needle aspiration of peritonsillar abscess in children. AB - Needle aspiration for the treatment of peritonsillar abscess was assessed in 43 consecutive children aged 7 to 18 years (mean age, 13.9 +/- 2.5 years) during the 3-year period from 1988 through 1991. A positive aspirate was obtained in 31 (76%) of the 41 patients who cooperated for needle aspiration; a mean of 2.9 +/- 1.9 mL of pus was withdrawn. Of the 31 children with a positive aspirate, in 27 (87%) the abscess resolved, two (6%) required a second aspiration for resolution, and two (6%) underwent immediate tonsillectomy for persistent abscess. Of the 10 children (24%) with negative aspirations, in six (60%) the abscess resolved with antibiotic treatment alone, three (30%) underwent immediate (quinsy) tonsillectomy, and in one (10%) the abscess spontaneously drained. No bleeding, airway obstruction, or anesthetic complications occurred. Needle aspiration of peritonsillar abscess in children, with tonsillectomy reserved for nonresponders, appears to be an efficacious and safe method of treatment. PMID- 8427680 TI - Wound tension in rhytidectomy. Effects of skin-flap undermining and superficial musculoaponeurotic system suspension. AB - This study was conducted to determine the effects of skin-flap undermining and superficial musculoaponeurotic system (SMAS) suspension on wound-closing tension. Nine sides from five fresh-frozen cadavers were used, with closing tension measured at the two main anchor points, anteriorly (A) and posteriorly (P), with and without SMAS plication for minimal (MIN), intermediate (INT), and maximal (MAX) skin-flap undermining. Results indicated that closing tension was significantly decreased with SMAS plication, both A and P, for all three levels of skin undermining. The average decrease in closing tension with SMAS plication was: A-MIN 191 g, A-INT 95 g, A-MAX 83 g, P-MIN 235 g, P-INT 68 g, and P-MAX 70 g (P < .001 for all). Considering the effect of skin-flap undermining alone, closing tension decreased with wider skin-flap undermining, both with and without SMAS plication. The tension-reducing effect of SMAS plication was decreased with wider skin-flap undermining. Regression analysis determined a second-order exponential curve relating closing tension to skin excision. PMID- 8427681 TI - Use of submentoplasty to enhance cervical recontouring in face-lift surgery. AB - A number of surgical techniques for recontouring the aging neck are available. Using liposuction and direct lipectomy, the surgeon can sculpt the jowl and submental contour. Posterior tightening of the ptotic platysma refines the submental area, while incisional or excisional techniques, along with midline plication, remove anterior banding. Despite these maneuvers, recurrent submental and cervical ptosis is noted in some patients. These individuals can benefit from a secondary submentoplasty, a submental tuck-up face-lift operation, usually within 6 to 18 months after their initial surgery. We have identified several reliable causes for a secondary submentoplasty. Submentoplasty as a planned second-stage surgery in certain patients allows achievement of more dramatic and long-lasting rejuvenation of the neck. PMID- 8427682 TI - Facial skeletal augmentation using hydroxyapatite cement. AB - This study investigates the use of a new calcium phosphate cement, which sets to solid, microporous hydroxyapatite, for facial bone augmentation. In six dogs, the supraorbital ridges were augmented bilaterally with this hydroxyapatite cement. On one side, the hydroxyapatite cement was placed directly onto the bone within a subperiosteal pocket. On the opposite side, the cement was contained within a collagen membrane tubule and then inserted into a subperiosteal pocket. The use of collagen tubules facilitated easy, precise placement of the cement. All implants maintained their original augmented height throughout the duration of the study. They were well tolerated without extrusion or migration, and there was no significant sustained inflammatory response. Histologic studies, performed at 3, 6, and 9 months revealed that when the cement was placed directly onto bone, progressive replacement of the implant by bone (osseointegration of the hydroxyapatite with the underlying bone) without a loss of volume was observed. In contrast, when the cement-collagen tubule combination was inserted, primarily a fibrous union was noted. Despite such fibrous union, the hydroxyapatite collagen implant solidly bonded to the underlying bone, and no implant resorption was observed. Hydroxyapatite cement can be used successfully for the experimental augmentation of the craniofacial skeleton and may be applicable for such uses in humans. PMID- 8427683 TI - Postoperative platysmal band deformity. A pitfall of submental liposuction. AB - Submental liposuction is an established advance in treatment of the aging neck. However, its misapplication may result in an aesthetic deformity involving postoperative anterior platysmal banding. To identify preoperative conditions relating to this sequela, we retrospectively analyzed 301 patients treated with submental liposuction. Submental obesity and an anatomic pattern of platysmal nondecussation were found to be significant correlates in the development of postoperative anterior platysmal banding. PMID- 8427684 TI - The coronal approach. Anatomic and technical considerations and morbidity. AB - The coronal flap has recently become a preferred approach for the otolaryngologist-head and neck surgeon requiring access to the craniofacial skeleton and orbit. The variety of cases in which it has proven indispensable include craniofacial reconstruction, facial trauma, and tumor resection. This method of exposure has become particularly useful with increased indications for rigid internal fixation and primary bone grafting in the management of complex facial fractures. Our experience is reviewed in terms of indications for and benefits of the coronal approach, with a detailed description of the technique emphasizing anatomic planes and neurovascular structures. Careful attention to the latter should allow prevention of potential complications. PMID- 8427685 TI - Dura mater for soft-tissue augmentation. Evaluation in a rabbit model. AB - Irradiated human dura mater, a commercially available preparation of dura mater (Tutoplast), and irradiated rabbit dura mater were implanted in subcutaneous pockets in the pinna and forehead of New Zealand white rabbits and evaluated for their usefulness in soft-tissue augmentation at 3 months and 6 months. Postoperatively, no evidence of erythema, purulence, hematoma or seroma formation, wound dehiscence, graft extrusion, or flap necrosis was noted. Irradiated human dura mater was well tolerated by the host and elicited a mild cellular inflammatory response. The graft was well preserved, infiltrated by fibrous connective tissue, and fixed in place in the pinna sites. Forehead grafts were surrounded by a capsule of host tissue and were intact. Dura mater seems to be useful for soft-tissue augmentation. Tutoplast may give good results clinically. Because this represents a short-term evaluation, long-term clinical results are needed to define the potential of dura mater grafts. PMID- 8427686 TI - The 11th nerve syndrome. Accessory nerve palsy or adhesive capsulitis? AB - The 11th nerve syndrome classically involves the majority of patients undergoing neck dissections even when the accessory nerve is preserved. A preliminary analysis of our data of 31 of 44 patients who underwent neck dissections from a prospective study showed numerous findings of shoulder disability that are not attributable to accessory nerve palsy but are well described by the syndrome of adhesive capsulitis of the glenohumeral joint. At 1 month postoperatively, although accessory nerve palsy symptoms were common, adhesive capsulitis symptoms were significant. At 6 months, the frequency of accessory nerve palsy symptoms was less as the accessory nerve had begun to recover. At 12 and 18 months, when most of the accessory nerves had recovered, the accessory nerve palsy symptoms were comparatively uncommon while the adhesive capsulitis symptoms predominated as the remaining symptoms of the 11th nerve syndrome. We propose that adhesive capsulitis is a principal component of the 11th nerve syndrome that can significantly compound the morbidity of a neck dissection even when the accessory nerve recovers. We also propose that adhesive capsulitis accounts for the persistence and variability of shoulder symptoms after neck dissection that cannot be attributed to trapezius muscle dysfunction. PMID- 8427687 TI - Facial nerve enhancement in Bell's palsy demonstrated by different gadolinium enhanced magnetic resonance imaging techniques. AB - Twenty-one patients with an acute complete peripheral facial palsy, Bell's palsy, were examined by medium- and high-Tesla magnetic resonance imaging. Three contrast techniques were used: intravenous gadolinium; oral carbohydrate and intravenous gadolinium; and gadolinium, carbohydrate, and readministration of gadolinium. Three to 22 days after the onset of palsy, 12 of the 21 patients demonstrated ipsilateral facial nerve enhancement, most consistently in the meatal region, which is indicative of an inflammatory reaction. Two to 4.5 months after the onset, the enhancement had disappeared in 10 of the 12 patients. For the individual patient, contrast-enhanced magnetic resonance imaging gave little or no help in predicting the outcome of palsy. It is speculated that the intake of carbohydrate and readministration of gadolinium may improve the sensitivity of medium-high-Tesla magnetic resonance imaging in some cases. PMID- 8427688 TI - Computerized slide show for the 1990s lecturer. AB - One of the most difficult and time-consuming tasks of writing and presenting papers is the review and coordination of patient photographic records. For years, not only has the 35-mm slide been the means of photographic storage for patient records, but also the standard for illustrating lectures and presentations. This current method is cumbersome and time consuming. We have used existing software of the Cameo II and imaging system to prepare, create, and present illustrations for lectures. Patient images are digitalized from any source (eg, photographs, slides, video tape) and electronically stored in the computer along with the computer-generated illustrations. When a lecture is prepared, the images are arranged sequentially in an electronic carousel. This computerized program is recorded on video tape and simply played through the video projector during the presentation to display the images. This system is more efficient since any stored image can be displayed instantaneously in any order without the handling of 35-mm slides. The only limitation of this system is the user's imagination. As the current trend in computer technology continues with high-resolution TV and video systems, the 35-mm slide presentation will become obsolete. PMID- 8427689 TI - Orbital decompression for preservation of vision in Graves' ophthalmopathy. AB - Graves' ophthalmopathy (thyroid eye disease) can result in progressive visual loss. The University of Washington (Seattle) experience in orbital decompression was reviewed for the years 1983 through 1990 to determine overall safety and outcome. Twenty patients underwent transantral decompression of 36 orbits for either steroid therapy failure, steroid therapy intolerance, or recurrence of optic neuropathy with tapering of the steroid therapy. Decompression successfully improved visual function in 33 of the orbits (92%) and a second decompression procedure was successful in another two (5%) of the orbits (6%). There were no major complications or cases of decreased visual function. Diplopia, present preoperatively in 17 patients (85%), was improved in eight patients (47%) and unchanged in nine patients (53%). However, of the three patients without preoperative diplopia (15%), one had development of new-onset diplopia postoperatively. Transantral decompression of the orbit offers a safe and effective therapeutic modality for vision-threatening Graves' ophthalmopathy. PMID- 8427690 TI - Water spout injection into the neck. AB - Work-related high-pressure injection injuries to the extremities are well publicized, and treatment plans are well established. However, water spout injection injuries to the oral cavity are rare. To our knowledge, there have been no reports of water injection injuries in the oral cavity. We describe one case in which a child suffered an intraoral low-pressure injury to the floor of the mouth, which resulted in water dissection to the neck and subcutaneous tissues. Management and eventual outcome are detailed. PMID- 8427691 TI - Pathologic quiz case 2. Chondrosarcoma of the maxilla. PMID- 8427692 TI - Pathologic quiz case 1. Cicatricial pemphigoid. PMID- 8427693 TI - Long-term outcome for children with autism who received early intensive behavioral treatment. AB - After a very intensive behavioral intervention, an experimental group of 19 preschool-age children with autism achieved less restrictive school placements and higher IQs than did a control group of 19 similar children by age (Lovaas, 1987). The present study followed-up this finding by assessing subjects at a mean age of 11.5 years. Results showed that the experimental group preserved its gains over the control group. The 9 experimental subjects who had achieved the best outcomes at age 7 received particularly extensive evaluations indicating that 8 of them were indistinguishable from average children on tests of intelligence and adaptive behavior. Thus, behavioral treatment may produce long-lasting and significant gains for many young children with autism. PMID- 8427695 TI - Teaching parents with mental retardation: knowledge versus skills. AB - An educational program for parents (N = 5) with mild mental retardation was evaluated to assess whether knowledge gains resulted in corresponding skill acquisition. Generalization of parenting skills to the home environment following behavioral skills training procedures was also examined. Parenting knowledge was assessed as subjects responded to descriptions of parenting situations. Parenting skills were assessed in home observations. Following baseline, a knowledge training procedure was implemented in a small group format. A behavioral skills training procedure was then implemented in a small group training format. Finally, individual behavioral skills training was provided in the home setting. Although knowledge training produced sizable increases in the knowledge measure, neither it nor the behavioral group training produced increases in parenting skills. However, following training in the home, each of the parenting skills increased substantially. The results were discussed with regard to the problem of generalization, and future research was proposed. PMID- 8427694 TI - Child-related and parenting stress: similarities and differences between mothers and fathers of children with disabilities. AB - Data concerning similarities and differences in child-related and parenting stress between mothers and fathers of 121 toddlers with disabilities were presented. Fathers reported more stress related to their child's temperament and their relationship to their child. Mothers reported more stress from the personal consequences of parenting. Differences between mothers and fathers regarding the most powerful predictors of child-related and parenting stress were also found. Fathers were more sensitive to the effects of the family environment, whereas mothers were more affected by their personal support networks. The implications for early intervention services were discussed. PMID- 8427696 TI - Patterns of leisure activities of young adults with mild mental retardation. AB - Leisure activities were examined for young adults with mild mental retardation. Some had disappeared from mental retardation services after leaving school; others were in adult day services. Peers without mental retardation (comparison subjects) were also included. Various leisure activities were combined using cluster analysis. Some adults with mental retardation and some comparison subjects were found in each of the 13 clusters obtained but proportions varied. Leisure patterns were strongly influenced by gender and marital status as well as mental retardation, except for one cluster, Socially Isolated, Impoverished, where there were more adults with mental retardation, irrespective of gender and marital status. PMID- 8427697 TI - Movement-related brain macropotentials of persons with Down syndrome during skilled performance. AB - Motor performances and movement-related brain macropotentials of nine 23-year-old males with Down syndrome whose mean MA was 10.2 years were compared to those of two control groups whose members were matched to the experimental subjects on CA or MA. A self-paced, voluntary, goal-directed task, which consisted of calculating a time interval of 40 to 60 msec by pressing two buttons, was employed. Subjects with Down syndrome had great difficulty in organizing and timing a correct temporal sequence of ballistic movements. They were much slower, less accurate, and achieved a smaller number of target performances. These behavioral patterns were associated with movement-related brain macropotentials altered in amplitude and latency. The potentials related to programming movements and processing reafferent sensory information were absent, and those involved in the evaluation of the outcome of the performance were reduced. PMID- 8427698 TI - Transition of localization of the N-Myc protein from nucleus to cytoplasm in differentiating neurons. AB - N-myc is a developmentally regulated proto-oncogene encoding a putative sequence specific DNA-binding protein. Previous studies on tissue distribution of N-myc transcripts indicated that one of the major sites of N-myc expression is the CNS and neural crest derivatives in developing embryos. We investigated N-Myc protein expression in embryonic neural tissues and found that the protein was usually localized in the nucleus, but accumulated in the cytoplasm upon differentiation of specific classes of neurons, e.g., retinal ganglion cells, neurons of spinal ganglia, and Purkinje cells of the cerebellum. The change of localization of N Myc from the nucleus to the cytoplasm indicates a novel feature of regulation of myc family proteins and suggests functions of N-myc in the cytoplasm of maturing neurons. PMID- 8427699 TI - Extensive dye coupling between rat neocortical neurons during the period of circuit formation. AB - A low molecular weight intracellular tracer, Neurobiotin, was injected into single neurons in living slices of rat neocortex made at postnatal days 5-18. Between days 5 and 12, 66% of single-neuron injections labeled clusters of up to 80 neurons surrounding the injected cell. Coupling between neurons occurred primarily through dendrites. Injections done in the presence of halothane, a gap junction blocker, abolished the spread of tracer to surrounding neurons, implying that gap junctions mediate coupling. Injections done after day 16 resulted in little or no dye coupling. We conclude that transient local coupling via gap junctions in developing cortex may provide a pathway for communicating intercellular signals, including subthreshold electrical activity, and thereby enable temporal coordination of local neuronal ensembles during circuit formation. PMID- 8427700 TI - Multiple calcium-dependent processes related to secretion in bovine chromaffin cells. AB - We have used the caged calcium compound DM-nitrophen to investigate the kinetics of calcium-dependent secretion in bovine chromaffin cells. Perfusion with partially calcium-loaded nitrophen often caused a loading transient--slow secretion for up to 1 min due to displacement of Ca2+ by cytoplasmic Mg2+. Flash photolysis elicited 100 microM [Ca2+]i steps that evoked intense secretion, lasting a few seconds. In cells experiencing a loading transient, [Ca2+]i steps evoked an especially fast secretion. A persistent, slow secretion often followed these fast phases. Distinct kinetic components may reflect secretion from pools that are differentially capable of release. Both secretion and movement of vesicles between pools appear to be [Ca2+]i sensitive. Later [Ca2+]i steps sometimes evoked a rapid capacitance decrease, indicating a fast, [Ca2+]i dependent phase of endocytosis. PMID- 8427701 TI - Ionotropic glutamate receptor subtypes activate c-fos transcription by distinct calcium-requiring intracellular signaling pathways. AB - N-Methyl-D-aspartate (NMDA) or non-NMDA receptor activation is sufficient to induce transcription of the immediate early gene c-fos in a calcium-requiring manner. We sought to determine whether the calcium-dependent mechanisms inducing c-fos transcription are identical following activation of these two receptor subtypes. We used in situ hybridization and fura-2 imaging to detect c-fos mRNA and intracellular calcium in individual dentate gyrus neurons maintained in vitro. Structurally distinct inhibitors of phospholipase A2 and cyclooxygenase abolished NMDA--but not kainic acid-induced increases of c-fos mRNA. Conversely, the calmodulin antagonist calmidazolium markedly inhibited kainic acid--but not NMDA-mediated increases of c-fos mRNA. We propose that the dissociation in the mechanisms transducing the calcium influx signals to the nucleus following NMDA and non-NMDA receptor activation is due to spatially distinct sites of calcium entry, resulting in activation of different enzymes located at distinct sites in the cell. PMID- 8427702 TI - A benzodiazepine recognition site associated with the non-NMDA glutamate receptor. AB - GYKI 52466 is a benzodiazepine molecule that has muscle relaxant and anticonvulsant properties not attributable to a gamma-aminobutyric acid receptor mediated mechanism. Here it is shown that GYKI 52466 exerts no blocking action at N-methyl-D-aspartate (NMDA) glutamate receptors, but acts noncompetitively to block ion currents and associated excitotoxicity, including ischemic neuronal degeneration, mediated through non-NMDA glutamate receptors. The inhibition of non-NMDA responses by GYKI 52466 is antagonized by cyclothiazide, hydrochlorothiazide, and diazoxide, benzothiadiazide drugs that inhibit non-NMDA receptor desensitization. These results suggest that non-NMDA receptor-ion channel complexes may contain a novel benzodiazepine recognition site where receptor desensitization is regulated; this postulated site represents a promising new target for rational development of drugs to treat neurological disorders. PMID- 8427703 TI - Low molecular weight elastase inhibitors in cells and tissues of alveolar regions: seek and ye shall find. PMID- 8427704 TI - Involvement of a signal transduction mechanism in ATP-induced mucin release from cultured airway goblet cells. AB - Release of mucins from cultured airway surface epithelial cells can be stimulated by extracellular ATP via a P2-purinergic receptor-mediated mechanism (K. C. Kim and B. C. Lee. 1991. Br. J. Pharmacol. 103:1053-1056). In this report, we studied the mechanism by which extracellular ATP induces the mucin release. We found that: (1) ATP increased both mucin release and generation of inositol phosphates in a dose-dependent fashion, and their dose-effect relationships were almost superimposed; (2) the increases in both mucin release and the phosphatidylinositol phosphate (PI) turnover by extracellular ATP were partially, but almost equally, blocked by the pretreatment with pertussis toxin (42% for mucin release and 44% for PI turnover). We conclude that in cultured airway goblet cells extracellular ATP stimulates mucin release by a signal transduction mechanism, which seems to involve coupling of ATP-activated P2 purinoceptors with phospholipase C, at least in part, via pertussis toxin-sensitive GTP-binding proteins. This may be an important finding in understanding the regulation of mucin release by airway goblet cells, since a number of agents present in the airway could influence this signal transduction pathway and subsequently modulate the mucin secretion. PMID- 8427705 TI - Secretion of mucus proteinase inhibitor and elafin by Clara cell and type II pneumocyte cell lines. AB - The regulation of proteinases secreted by neutrophils is very important for the prevention of tissue injury. We recently described the isolation of elafin from bronchial secretions, a new elastase-specific inhibitor that is also found in the skin of patients with psoriasis. In this study, we investigated the secretion of elafin and mucus proteinase inhibitor (MPI), another inhibitor showing sequence similarity with elafin, in two lung carcinoma cell lines, NCI-H322 and A549, which have features of Clara cells and type II alveolar cells, respectively. The results presented show that the two inhibitors are produced when the cells are cultured either in serum-free or in serum-containing media. MPI was detected immunologically as a unique molecule of M(r) 14 kD, in accordance with previous studies. Conversely, one or two elafin-immunoreactive species were detected depending on the cell line: a 12- to 14-kD species was observed in the A549 cell line, regardless of the culture conditions, whereas in the NCI-H322 cell line we detected a 6-kD species in serum-containing (10% fetal calf serum) conditions and a 12- to 14-kD species in serum-free conditions. The 12- to 14-kD molecule probably represents an active precursor of elafin. Whether the cleavage of the 12 to 14-kD precursor giving rise to the elafin molecule is of any physiologic significance is not known. In showing for the first time that MPI and elafin (and its precursor) are secreted by the A549 cell line, this report implicates the type II alveolar cell in the defense of the peripheral lung against the neutrophil elastase secreted during inflammation. PMID- 8427706 TI - Cell surface carbohydrates of rat alveolar type II cells in primary culture. AB - Cell surface carbohydrates have been shown to be altered during cellular differentiation. Alveolar type II (ATII) cells in culture gradually lose their differentiated phenotype. Therefore, the aim of this study was: (1) to characterize changes in terminal carbohydrates of cell surface glycoproteins of rat ATII cells cultured for 1 to 5 days on plastic, and (2) to assess the concomitant changes in sialidase and sialyltransferase activity of ATII cell homogenates. Cells were surface-labeled with potassium-[3H]-borohydride after oxidation by sodium periodate at millimolar concentrations, galactose oxidase or neuraminidase plus galactose oxidase, allowing for the specific labeling of terminal sialic acids, terminal galactose/N-acetylgalactosamine (Gal/GalNAc), or terminal an penultimate Gal/GalNAc residues, respectively. Glycoproteins were separated by SDS-PAGE. On day 1, cells were heavily coated with sialic acids, since no labeling could be introduced with galactose oxidase alone. From day 1 to day 5, we observed a selective and progressive desialylation of two glycoproteins (200 and 165 kD). At the same time, the ATII cells' sialidase activity (pH 4.2) exhibited an 8-fold increase (60.3 +/- 4.0 pmol/min/mg protein on day 1 versus 406.9 +/- 3.7 pmol/min/mg protein on day 5), whereas the sialyltransferase activity increased 2-fold (212 +/- 8 fmol/min/mg protein on day 1 versus 395 +/- 82 fmol/min/mg protein on day 5) and the supernatant sialidase activity was unchanged (2.8 +/- 0.7 pmol/min/ml on day 5). Thus, the phenotypic changes of ATII cells in primary culture are accompanied by a partial cell surface desialylation and an increase in intracellular sialidase activity. PMID- 8427707 TI - Regulation of interleukin-1ra, interleukin-1 alpha, and interleukin-1 beta production by human alveolar macrophages with phorbol myristate acetate, lipopolysaccharide, and interleukin-4. AB - Human alveolar macrophages (AM) are antigen-presenting cells that have an important immune effector function in the lung. We have previously shown that AM produce a specific interleukin-1 (IL-1) inhibitor of 20 to 25 kD that blocks biologic activities of IL-1 alpha and IL-1 beta such as prostaglandin E2 production by fibroblasts. This inhibitor acts as a receptor antagonist (IL-1ra) by binding to the IL-1 receptor. We are now presenting evidence that the natural AM-derived IL-1ra is immunologically identical to IL-1ra cloned from human peripheral blood monocytes and shows a band at 20 kD compatible with the natural glycosylated IL-1ra. No constitutive expression of IL-1 mRNA was detected when analyzed by Northern blot immediately after bronchoalveolar lavage from six control patients. Comparison of in vitro kinetics of IL-1ra, IL-1 alpha, and IL-1 beta analyzed during culture in the presence or absence of phorbol myristate acetate revealed that their mRNA expression was asynchronous. IL-1 alpha and IL-1 beta mRNA were expressed after as little as 15 min, whereas IL-1ra mRNA was detectable only after 3 h in culture. The production of IL-1ra was measured by enzyme-linked immunosorbent assay and compared with that of IL-1 alpha and IL-1 beta. In freshly isolated AM (10(6)/ml), cell-associated IL-1ra was present in an average amount of 2.0 +/- 0.5 ng/ml, i.e., 25 and 100 times more than IL-1 alpha and IL-1 beta, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427708 TI - Muscarinic receptor subtypes in human nasal mucosa: characterization, autoradiographic localization, and function in vitro. AB - Muscarinic receptors play important roles in the regulation of glandular secretion and vasomotor tone in human nasal mucosa. M1, M2, and M3 muscarinic receptor subtypes were pharmacologically characterized in human inferior turbinates by receptor-binding assays using [3H](-)quinuclidinyl benzilate (QNB, identifies total muscarinic receptors) and [3H]-pirenzepine (PZ). Receptors were localized by autoradiography, and their function examined in vitro by assaying mucus secretion from cultured nasal mucosal explants. In competition assays, PZ was employed as a selective muscarinic antagonist for M1 receptors, gallamine and AF-DX 116 for M2 receptors, and 4-DAMP for M3 receptors. These ligands are selective at low nanomolar concentrations, but can interact with other muscarinic receptors at higher concentrations. It is not known if they can interact with putative M4 and M5 muscarinic receptor subtypes. Using [3H](-)QNB, total muscarinic receptor binding was 688.4 +/- 49.6 fmol/mg protein (Bmax), with a Kd of 1.47 +/- 0.13 nM. [3H]-PZ bound to 45% of the total QNB binding sites. In competition experiments, 4-DAMP displaced [3H](-)QNB with the lowest IC50, followed by PZ and AF-DX 116. Autoradiograms demonstrated that [3H](-)QNB binding was completely displaced by 4-DAMP, partially displaced by PZ, but not displaced by gallamine or AF-DX 116, and suggested that M1 and M3 subtypes coexist in submucosal glands. The localization of M1 receptors on submucosal glands was confirmed by direct labeling with [3H]-PZ. [3H]-PZ also labeled vessels, but with a low silver grain density. Autoradiographic [3H]-QNB binding was displaced by 4 DAMP and atropine, but not by PZ, gallamine, or AF-DX 116. In studies of mucus secretion in vitro, 4-DAMP significantly inhibited methacholine-induced secretion. Although less effective, PZ also had significant inhibitory effects. Neither gallamine nor AF-DX 116 had any inhibitory effect. M1 receptors (PZ binding sites) may regulate glandular secretion while M3 receptors (4-DAMP binding sites) may regulate glandular secretion and vasomotor tone in human nasal mucosa. PMID- 8427709 TI - Attenuated fibroblast sheath around the basement membrane zone in the trachea. AB - In the connective tissue underlying the epithelium of the airways, there are thin attenuated cells that appear to be fibroblasts. These cells are close to the basal lamina and delineate an area equivalent to the basement membrane zone (BMZ). Using the mean data from transversely and longitudinally sectioned attenuated fibroblasts, we estimate that the diameter of the attenuated fibroblast is approximately 28.0 microns and its thickness 0.55 microns. Serial sections indicate there are pores of various sizes in the attenuated fibroblast. Also, the presence of cell segments less than 5 microns in length suggests there are processes from the edge of the cell which would give it a stellate appearance. These large flat cells form a sheath covering 66 to 70% of the BMZ at an average distance of 1.9 microns from the basal lamina. In addition, they make contact with the basal lamina approximately 7,000 times per mm2 of basal lamina. The majority of the contacts with the basal lamina are beneath basal cells. Cells of the attenuated fibroblast sheath are probably associated with lung morphogenesis, wound healing, and development of the BMZ. Their existence as a sheath demonstrates an anatomical unit associated with fibroblast-epithelial cell interactions in airway epithelium. PMID- 8427710 TI - Defensins reduce the barrier integrity of a cultured epithelial monolayer without cytotoxicity. AB - Polymorphonuclear leukocytes (PMN) contribute to epithelial injury at sites of inflammation, but their mechanisms of action are incompletely understood. PMN can injure target tissues by oxidative and nonoxidative mechanisms. Included in the nonoxidative mechanisms are defensins (DEF), small (3.5 to 4.0 kD), arginine- and cysteine-rich polypeptides. DEF are bactericidal, fungicidal, viricidal, and tumoricidal, but their ability to contribute to inflammatory injury has not been extensively evaluated. One marker of inflammatory injury is disrupted epithelial barrier integrity. Using Madin-Darby canine kidney (MDCK) epithelial monolayers, we measured the effect of both human and rabbit DEF on barrier integrity using mannitol permeability (Pmann) and transepithelial electrical resistance (Rt). Human DEF (HNP1-3, 2:2:1 molar ratio) increased Pmann in a time- and concentration-dependent manner and Rt fell progressively over a 48-h period after exposure of monolayers to HNP1-3. Rabbit DEF peptide 1 (NP-1) also increased Pmann, but rabbit peptide 5 (NP-5) had no effect on Pmann. To investigate the role of charge, HNP1-3 was added to the monolayers with the polyanions heparin or sulfated dextran. Heparin and sulfated dextran only partially inhibited the increase in Pmann. Fetal bovine serum (FBS), however, completely inhibited the effect of HNP1-3, but this protection was only partially explained by the anionic protein, albumin. The FBS protection was time dependent and was present when FBS was added up to 16 h after exposure to HNP1-3. While both HNP1-3 and NP-1 increased epithelial permeability, neither were cytolytic to MDCK cells as measured by lactate dehydrogenase release.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427711 TI - Tumorigenic conversion of human mesothelial cells as a consequence of platelet derived growth factor-A chain overexpression. AB - Overexpression of platelet-derived growth factor (PDGF)-A as well as PDGF-B chain mRNA has previously been reported in human mesothelioma cell lines. In this report, it has been established that the A but not the B chain protein is expressed at detectable levels in cell lysates and conditioned medium from these cell lines. In order to investigate the effect of overexpression of PDGF-A chain in a human mesothelial cell model system, a retroviral vector containing a human PDGF-A chain cDNA insert under the control of the Moloney murine leukemia virus (MoMLV) promoter was inserted into the SV-40 T-antigen immortalized human mesothelial cell line MeT-5A. Selected cells showed overexpression of PDGF-A chain relative to MeT-5A and induced tumors in athymic nude mice. PDGF-A chain overexpression was also found in the tumor specimens excised from the mice. PDGF A mRNA and protein were expressed at a higher level in the tumor explant cell lines, suggesting a correlation of tumorigenicity with A chain production. PMID- 8427712 TI - Differential glucocorticoid regulation of the pulmonary hydrophobic surfactant proteins SP-B and SP-C. AB - Glucocorticoids increase expression of the genes for the pulmonary surfactant associated proteins SP-B and SP-C in fetal lung both in vivo and in vitro. To examine the mechanism of these effects, we studied induction of SP-B and SP-C mRNAs in human fetal lung cultured as explants. Both mRNA levels rose rapidly in response to 100 nM dexamethasone (Dex), with a faster response for SP-B: maximal levels of induction were achieved in < or = 12 h for SP-B (3.5-fold versus control) versus approximately 24 h for SP-C mRNA (35-fold versus control). Cycloheximide (2.5 micrograms/ml) did not affect glucocorticoid induction of SP-B mRNA but markedly decreased induction of SP-C mRNA. In control cultures, cycloheximide did not significantly reduce levels of either transcript. In nuclear run-on assays, Dex increased the rate of gene transcription for both SP-B (2.8 +/- 0.3-fold versus control, n = 4) and SP-C (10- to 30-fold). Using actinomycin D to assess mRNA stability, the t1/2 of SP-B mRNA was increased from 7.5 +/- 0.4 h to 18.8 +/- 2.9 h by Dex treatment (P < 0.05), whereas the t1/2 of SP-C mRNA was not affected (9.3 +/- 1.7 h versus 8.1 +/- 1.2 h; NS). A similar increase in SP-B mRNA t1/2 with Dex (from 6 h to 19 h) was observed in label chase studies with [3H]uridine. We conclude that glucocorticoids regulate the hydrophobic surfactant proteins of alveolar type II cells by different mechanisms: induction of SP-B is a primary response and includes an increase in both transcription rate and mRNA stability, whereas induction of SP-C is a secondary process, requiring ongoing protein synthesis, involving increased transcription rate without a change in mRNA stability. PMID- 8427713 TI - Qualitative changes in the human T-cell leukemia/lymphotropic virus type I env gene sequence in the spastic versus nonspastic tropical paraparesis are not correlated with disease specificity. PMID- 8427714 TI - Studies on the specificity of the vaccine effect elicited by inactivated simian immunodeficiency virus. AB - Inactivated, partially purified simian immunodeficiency virus (SIVmac) protected macaques from intravenous challenge with homologous and heterologous strains of SIV that had been grown on human cells but no protection against challenge with monkey peripheral blood mononuclear cell-grown SIVmac was afforded. Human immunodeficiency virus type 1 prepared in an analogous way to the SIVmac vaccine on the C8166 human T cell line protected macaques against challenge with human cell-grown SIVmac. These results suggest that protection may be mediated by xenoimmunization with the vaccine cell substrate proteins. All vaccinated macaques had anti-cell antibodies. Major reactivity to MHC class I antigens was found as well as to a 70-kD protein detectable only under nonreducing conditions. PMID- 8427715 TI - Human immunodeficiency virus type I provirus is demonstrated in peripheral blood monocytes in vivo: a study utilizing an in situ polymerase chain reaction. AB - Human immunodeficiency virus type 1 (HIV-1) infects a variety of cell types in vivo. Monocyte/macrophages may be the major reservoir for HIV-1 in the solid tissues of HIV-1-infected individuals. Conflicting data have been reported, though, regarding the presence of HIV-1 provirus in peripheral blood monocytes isolated from HIV-1-seropositive humans. We have evaluated monocytes from the peripheral blood of eleven HIV-1-infected individuals utilizing a new, highly sensitive and specific in situ polymerase chain reaction. We demonstrate HIV-1 provirus in 73% (8/11) of these samples. None of these monocyte samples was demonstrated to contain cells expressing high levels of HIV-1-specific RNA, by standard in situ hybridization. The evaluation of the HIV-1 genome in peripheral blood monocytes of certain infected individuals may assist in the understanding of HIV-1 proviral latency and pathogenesis, in vivo. PMID- 8427716 TI - Human immunodeficiency virus type 1 RNA detection in peripheral blood mononuclear cells by polymerase chain reaction: enhanced sensitivity after mitogenic stimulation. AB - The aim of this study was to investigate whether stimulus-induced up-regulation of human immunodeficiency virus type 1 (HIV-1) expression in peripheral blood mononuclear cells (PBMC) could enhance the diagnostic sensitivity of the polymerase chain reaction (PCR). PBMC derived from 11 HIV-1-infected asymptomatic adults were cultured with a stimulus of phytohemagglutinin (PHA) plus phorbol 12 myristate 13-acetate (PMA) for 36 h prior to lysing the cells for PCR. In all 11 patients studied, the intensity of PCR-assisted HIV RNA amplification (RNA-PCR) performed on stimulated cells was significantly (p < 0.001) higher than that obtained on unstimulated cells. A comparison of conventional PCR-assisted DNA amplification (DNA-PCR) with that of RNA-PCR was made on seven patients. The sensitivity of DNA-PCR was also increased by prior stimulation of cells, although not to the same extent as was observed for RNA-PCR. The results of our study indicate that the sensitivity of PCR can be significantly enhanced by prior activation of cells with PHA and PMA. PMID- 8427717 TI - Inhibition of human immunodeficiency virus type 1 replication by hydroxychloroquine in T cells and monocytes. AB - Chloroquine and its analogue hydroxychloroquine (HCQ) have been shown to inhibit a variety of viral infections including influenza and adenovirus through blockade of viral entry via inhibition of endosomal acidification. We have extended these observations to human immunodeficiency virus type 1 (HIV-1) infection utilizing primary T cells and monocytes, a T cell line (CEM), and a monocytic cell line (U 937). HCQ inhibited HIV-1 replication (> 75%), as measured by reverse transcriptase activity, in the primary T cells and monocytes as well as the T cell and monocytic cell lines. HCQ itself had no anti-reverse transcriptase activity and was not toxic to the cells at concentrations inhibitory to viral replication. Intracytoplasmic staining with an anti-p24 antibody, 24 h after infection, revealed the presence of intracytoplasmic virus, suggesting that the drug does not block viral entry. The production of steady-state HIV-1 mRNA was not affected by HCQ in that comparable levels of HIV-1 mRNA could be detected by Northern blot analysis and by in situ hybridization in both the HCQ-treated and untreated cells. However, HCQ does appear to affect production of infectious HIV 1 virions because viral isolates from HCQ-treated cells could not infect target CEM cells. These data suggest that HCQ may be useful adjunctive therapy in the treatment of HIV-1 infection. PMID- 8427718 TI - The epidemic of human immunodeficiency virus infection in Romanian children. AB - Between June 1989 and May 1991, 29,020 subjects were tested for anti-human immunodeficiency virus (HIV) antibodies; 1079 were seropositive. A big discrepancy existed between the high seroprevalence in the group 0-3 yr of age, 11.22%, and that of 0.43% in the adult population. Moreover, only a small percentage of the pediatric cases (4%) resulted from maternal infection. These observations indicate viral spread by horizontal transmission. Blood, blood products, and reusable unsterilized needles and syringes were probably the main route by which HIV spread in the infant population in medical institutions. The initial source of HIV infection in Romania remains to be determined. PMID- 8427719 TI - The 5-hydroxytryptamine receptor antagonists as antiemetics: preclinical evaluation and mechanism of action. PMID- 8427720 TI - Ondansetron. AB - Ondansetron is the first selective antagonist of the 5-hydroxytryptamine receptors (type 3) marketed for the prevention of emesis induced by antineoplastic agents. Ondansetron has been shown to be more active and less toxic than high-dose metoclopramide in patients submitted to cisplatin chemotherapy. Furthermore, when dexamethasone was added to ondansetron, its antiemetic efficacy increased significantly. In the prevention of emesis induced by a high single dose of cisplatin or by repeated low doses, ondansetron combined with dexamethasone has been shown to be the more efficacious and less toxic antiemetic treatment. However, in the prevention of delayed emesis from cisplatin, its role is still to be defined. In patients submitted to moderately emetogenic chemotherapeutic agents, ondansetron has shown an efficacy superior or equal to standard doses of metoclopramide, but is less toxic. Moreover, when compared with dexamethasone, its antiemetic efficacy and tolerability is similar; in this group of patients ondansetron should be used only when steroids fail. Ondansetron toxicity is generally mild; in particular, it does not induce extrapyramidal reactions. The most frequent side-effects are headache and constipation. PMID- 8427721 TI - The new fluorinated quinolones for antimicrobial prophylaxis in neutropenic cancer patients. AB - Fluoroquinolones are the most attractive agents for prophylactic use in neutropenic cancer patients, due to their broad antimicrobial spectrum, high concentration in the faeces, systemic bactericidal activity, uncommon emergence of resistant strains and good tolerability. They have proved to be more effective than placebo, oral non-absorbable antibiotics or cotrimoxazole in the prevention of Gram-negative infections. In a prospective, randomised multicentre study performed by the GIMEMA infection program, ciprofloxacin was demonstrated to be more effective than norfloxacin for the reduction of febrile episodes, use of systemic antibiotics, and Gram-negative infections in neutropenic patients with haematological malignancies. The greater efficacy may be related to its better systemic or greater antibacterial activity. The potential problems related to the prophylactic use of fluoroquinolones are the increasing prevalence of Gram positive infections caused by streptococci and coagulase-negative staphylococci; the reported emergence and nosocomial spread of resistant strains, especially among coagulase-negative staphylococci; the lack of their usefulness as empirical therapy in febrile neutropenic patients. Fluoroquinolones are today the better choice for preventing Gram-negative infections in neutropenic patients and ciprofloxacin should probably be preferred. More information on their efficacy and their relationship to the overall susceptibility of micro-organisms in patients with cancer would be valuable, and careful monitoring of patients treated with these drugs is therefore warranted. PMID- 8427722 TI - Tropisetron (ICS 205-930): a selective 5-hydroxytryptamine antagonist. PMID- 8427723 TI - LAS 30451: a novel 5-HT3 antagonist. PMID- 8427724 TI - Delayed emesis following high-dose cisplatin: a double-blind randomised comparative trial of ondansetron (GR 38032F) versus placebo. AB - Despite recent advances in control of acute emesis, delayed nausea and vomiting following cisplatin-based chemotherapy remain a significant cause of treatment related morbidity. Ondansetron, a selective 5HT3 receptor antagonist, is effective in preventing acute emesis in the initial 24-h period following high dose cisplatin. The efficacy and safety of ondansetron in preventing the delayed emesis syndrome during days 2-5 after cisplatin (> or = 100 mg/m2) were evaluated in a double-blind, placebo-controlled multicentre trial. 50 patients having two or fewer emetic episodes during the first 24 h were randomised to receive ondansetron (16 mg) or placebo orally three times daily beginning 24 h after cisplatin. Rates of complete control of emesis were higher in ondansetron-treated patients during each study day, 59-78%, compared with 39-50% in placebo-treated patients, but the differences were statistically superior only on the third study day (P = 0.009). 40% of patients in the ondansetron treatment arm and 33% treated with placebo had complete control of emesis during the entire 4-day study period (P = 0.648). Withdrawal from study due to nausea and vomiting occurred in 13% of ondansetron-treated patients compared with 33% in the placebo arm (P = 0.102). Control of nausea was better with ondansetron, but differences were not statistically significant. Adverse effects of oral ondansetron given in this dose schedule were minimal. These data suggest that the delayed emesis syndrome may be partially mediated through the 5HT3 receptor, but that a serotonin antagonist alone provides inadequate control. Further investigation of ondansetron-based therapy in this clinical setting is warranted. PMID- 8427725 TI - Potential for combination therapy with the new antiserotonergic agents. AB - Appreciation of the major role played by serotonergic (5-HT3) neuroreceptors in the emetic reflex arc has introduced an additional factor into the rational design of combination antiemetic therapy. Combinations of an antidopaminergic agent and a corticosteroid have previously served as the basis for many successful antiemetic regimens. Three pilot studies and three randomised studies have now demonstrated potentiation of antiemetic activity of a 5-HT3 antagonist by dexamethasone as well. Further development of combination antiemetic regimens may involve antagonism of additional receptors including those for benzodiazepines, opiates, and catecholamines. Even antidopaminergic agents may continue to have a role. Although high-dose metoclopramide has both antiserotonergic and antidopaminergic activity, other pure antidopaminergic agents retain significant antiemetic activity. The combination of an antiserotonergic agent and a low-dose antidopaminergic agent has already shown promise in one pilot study. Newer and more effective antiemetic combinations will be needed to continue to improve the quality of life of patients receiving chemotherapy. PMID- 8427726 TI - Quality of life in breast cancer patients: the contribution of data to the care of patients. AB - A primary test of the usefulness of quality of life research is the extent to which it has been helpful in informing clinical practice and in minimising or preventing psychosocial distress among patients and their families. Clinical applications of quality of life data in five areas are summarised: alerting physicians and nurses to common patient concerns, informing patients of common reactions to breast cancer, aiding patients and physicians in decision-making, developing training programmes for medical personnel, and designing interventions for patients and their families. Although substantial progress has been made in applying quality of life data to improve clinical practice, further progress is possible. As with many other areas of medical practice, we must work towards developing more effective means of disseminating this information to clinicians and encouraging them to integrate it into their practice. PMID- 8427727 TI - Empirical antibiotic therapy in neutropenic cancer patients. PMID- 8427728 TI - Reliability and validity of a quality of life questionnaire in cancer patients. AB - Two studies were sequentially conducted to validate a new questionnaire which takes into consideration the most important variables which could influence quality of life evaluation. Particular attention was given to the methodology employed to collect data and to the patients' characteristics. In the first study 80 consecutive cancer patients were randomised to twice fill in one of four different types of questionnaire, each one characterised by a different polarisation of semantic and syntactic extreme values of the visual linear analogue (for instance, "very much" always on the right, regardless of the semantic value of the answer; positive semantic value always on the right, regardless of whether it was "very much" or "not at all"; and so on). The second study, conducted on 60 lung cancer patients, consisted in testing the reliability (by measuring the reproducibility in different ways) and the validity (by performing a factor analysis) of the type of questionnaire indicated by the first study as the most reliable. The internal coherence was also evaluated by measuring the effects of physical and psychological conditions on the responses. PMID- 8427729 TI - Current classifications of HIV-associated periodontal diseases. AB - The occurrence of unusual and severe forms of periodontal disease in HIV-infected individuals is well recognised. Several classification schemes have been proposed in an effort to associate periodontal deterioration with progressive stages of HIV infection and to determine aetiological factors in tissue destruction. No classification to date has proved entirely satisfactory. This paper reviews current classification schemes and suggests an alternative based upon the periodontal status of a cohort of HIV seropositive patients in Edinburgh. The proposed classification includes 'conventional', non-specific gingivitis and adult periodontitis such as occur in HIV seronegative individuals, but which may also be seen in seropositive subjects. However, conditions termed linear gingivitis and necrotising periodontitis may be pathognomonic for HIV infection. It is recommended that the term 'HIV-associated' be dropped in relation to periodontal disease. PMID- 8427730 TI - A survey of patients seen in consultant clinics in conservative dentistry at Edinburgh Dental Hospital in 1990. AB - A baseline study of referrals by letter to a department of conservative dentistry staffed purely by honorary consultants was conducted for one year. Distinct differences were found between the majority of patients referred from general practice and those referred from other sources. Principal reasons for referral were problems of tooth replacement, appearance, endodontics and tooth wear. A predominance of female patients was seen in all groups except those complaining of tooth wear. 'Endodontic' patients featured the highest proportion of requests for hospital treatment. Complex restorative treatment was provided for the majority (70%) of referred patients, and about 10% received treatment from dental students. The rates of referral showed evidence of dependence on travelling distance to the referral centre but not on the availability of local GDPs. This emphasises the importance of maintaining existing specialist dental referral services in major population centres and the development of new services where appropriate. PMID- 8427731 TI - Dental education in Japan. AB - Expanded roles for chairside assistants, infrequent use of amalgam, and dental technician courses over-subscribed by three to one. Perhaps this isn't the classic image of the advanced society that Japan has become over the last 40 years, yet the dentists working and researching in this economic spearhead were full of enthusiasm for their profession. On our journey to the far east we would be greeted with many surprises, especially in dental practice and hygienist education. PMID- 8427732 TI - Drenched in symbolism. AB - The choosing of a logo is probably one of the most fraught exercises in the history of the world. Certainly far more taxing than a large pinned amalgam in an upper left eight, infinitely more irritating than the seventeenth ease on the same denture and, frankly, enough to turn the most fervent democrat into a rabid dictator. PMID- 8427733 TI - Emergency dental services. PMID- 8427734 TI - The third molar epidemic. PMID- 8427735 TI - Mike Grace talks to John Murray. PMID- 8427736 TI - Rampant caries--a new threat? PMID- 8427737 TI - Recovery position. PMID- 8427738 TI - 'The orthodontic maze'. PMID- 8427739 TI - The orthodontic maze. PMID- 8427740 TI - Whistle blowing: the ethics of revealing professional incompetence within dentistry. AB - Traditionally, the dental profession has endorsed the view that one member should not make public--to patients or colleagues--criticisms of the work of another. Recently, the General Medical Council has disagreed, stating that there may well be circumstances when doctors should not protect colleagues from the consequences of their own mistakes. In this paper we endorse this view. We argue that 'whistle blowing' is appropriate in dentistry as well when its aim is to protect patients from the serious harm to which ignoring such errors might lead. PMID- 8427741 TI - MR lymphography using iron oxide particles. Detection of lymph node metastases in the VX2 rabbit tumor model. AB - MR images of the iliac lymph nodes of 25 VX2 carcinoma-bearing rabbits and of 5 tumor-free rabbits were obtained at 1.5 T before and after endolymphatic administration of superparamagnetic iron oxide particles (SPIO) at a dose of 1 mumol Fe per extremity. Imaging results were correlated with histology. In unenhanced images intranodal metastases were not detectable with any of the pulse sequences used and the signal intensities of tumor-free and metastatic lymph nodes did not differ significantly. After administration of the contrast medium, a significant signal loss (p < or = 0.05) occurred in the healthy lymph node tissue, whereas the signal intensity of lymph node metastases remained unchanged. In SPIO enhanced images, the threshold size for detection of lymph node metastases was 2 mm. Metastatic involvement was detected in 28 of the 30 tumorous lymph nodes with the SE 2000/15 sequence but in a smaller number of lymph nodes with the sequences SE 500/22 (n = 27) and 2000/65 (n = 21). PMID- 8427742 TI - Functional MR imaging at 1.5 T. Initial results using photic and motoric stimulation. AB - A preliminary investigation of the effects of stimulation of the visual and the motor cortex was made on a conventional 1.5 T MR imaging scanner. Both types of activation gave a detectable change in the signal between rest and stimulation using a gradient echo sequence with an echo time of 60 ms. The observed effects were assumed to be caused by variation in the amount of paramagnetic deoxyhemoglobin between stimulation and rest due to local increase of capillary blood flow in the human brain during stimulation. PMID- 8427743 TI - MR relaxation times and fiber type predominance of the psoas and multifidus muscle. An autopsy study. AB - MR relaxation times, fiber composition, nonmyofiber space, water content, and fat content of human psoas and multifidus muscle samples of 10 male cadavers were studied in vitro. The T1 and T2 relaxation times of multifidus muscle were significantly longer than those of the psoas muscle. On average, type 1 fibers (slow fibers with a small cross-sectional diameter) predominated in both muscles. There was no correlation between the relative mass of type 1 or 2 fibers (fast fibers with a large cross-sectional diameter) or nonmyofiber space and the relaxation times. The quantity of fat in the muscle did not correlate with the relaxation times either. PMID- 8427744 TI - Quantification of inhomogeneities in malignancy grading of non-Hodgkin lymphoma with MR imaging. AB - In a previous study of 50 patients with non-Hodgkin lymphoma (NHL) it was shown that the inhomogeneous appearance of a tumor at MR imaging strongly indicated a high malignancy grade. In this study of 33 patients with NHL, the administration of an i.v. contrast medium, Gadolinium-DTPA, improved the subjective detectability of the inhomogeneities. A method of quantifying the degree of inhomogeneity in the tumors (inhomogeneity index, IH-index) was developed and tested. The mean value of IH-index in the T2-weighted image before contrast medium administration, and of the T1-weighted image after contrast medium administration, as well as the IH-index value in the T2-weighted image before contrast medium administration alone, was able to discriminate well between low- and high-grade NHL. This method of quantifying the degree of inhomogeneity in tumors improved sensitivity in detecting high-grade NHL. PMID- 8427745 TI - Percutaneous ultrasound-guided biopsy. Fine needle biopsy, cutting needle biopsy, or both? AB - The results of 155 US-guided cutting needle biopsies, mainly of abdominal and transthoracic lesions, were reviewed to assess the diagnostic accuracy of the method. A fine needle biopsy (FNB) was additionally performed in 99 of the patients. Sufficient material for histologic and cytologic analysis was obtained in 88% (137/155) and 98% (97/99), and a correct benign or malignant diagnosis was made in 87% and 88%, respectively. Among the 99 combined biopsies the corresponding figures were 100% and 97%, respectively. The proportions of inconclusive and false-negative results among histologic samples were 0% and 4% (2/57), in FNBs 7% (7/99) and 5% (3/56), and in combined examinations 3% (3/99) and 0%. One major complication, Streptococcus faecalis sepsis, occurred. The combined use of FNB and histologic biopsy increases the proportion of correct diagnoses about 10% without increasing the complications. PMID- 8427746 TI - Anal endosonographic findings in patients with obstructed defecation. AB - Anal endosonography, including measurements of anal sphincter size, was performed in 16 patients with obstructed defecation. The findings were compared with those at defecography and anal manometry. Patients with rectocele and intussusception had a normal endosonographic appearance. One patient with puborectalic spasm had normal sonography. There was no correlation between sphincter size and anal manometry. The external sphincter muscle was thicker and the cross-sectional area larger in patients with obstructed defecation than in healthy controls (p < 0.05). Two patients with sphincter spasm and impaired rectal emptying at defecography had clearly thickened internal sphincters which may be the cause of their defecatory disorder. Three patients with previous anal dilatation or hemorrhoidectomy had sphincteric defects. Anal endosonography may be considered in patients with obstructed defecation to identify patients with internal sphincter hypertrophy. PMID- 8427747 TI - Inability of refined CT to assess local extent of prostatic cancer. AB - A refined procedure for CT was evaluated as a staging procedure in 19 patients with localized prostatic carcinoma. The CT images were compared to histopathologic whole-mount step sections of the surgical specimens. Fourteen of the patients had pathologic stage T3 (pT3) and 5 had stage pT2 giving a prevalence of extracapsular growth of 0.74. The CT images were read by 2 radiologists independently with a diagnostic accuracy of 0.37 for both observers. This is no better than in previous studies using a routine CT procedure. We conclude that CT is of little value in the staging protocol for the local extent of prostatic carcinoma before radical prostatectomy. PMID- 8427748 TI - Radial scar and tubular carcinoma. Mammographic and sonographic findings. AB - Clinical, mammographic, sonographic and pathologic examinations in 17 patients with nonpalpable stellate lesions showed radiographic findings suggestive of radial scars, with the exception of 2 cases which showed a dense central region. These findings were not sufficiently consistent to differentiate radial scar from carcinoma so that wire localization biopsy was necessary. Four patients had 7 radial scars associated with 6 tubular carcinomas, 2 of them having 3 and 2 radial scars with 2 tubular carcinomas, respectively. No characteristic radiographic findings were made in these cases. Four patients showed radial scar with punctate microcalcifications in the surrounding breast parenchyma as part of benign and premalignant conditions. Ultrasonography in 7 patients showed a hypoechoic area on 4 with acoustic shadow similar to that of carcinoma. Our study suggested that radial scar has a relationship with tubular carcinoma and surgical excision of these stellate lesions is, therefore, required. PMID- 8427749 TI - Reference values for vertebral heights in Scandinavian females and males. AB - Vertebral heights were measured on lateral spine radiographs covering T4-L5 in 113 healthy Caucasian volunteers (73 females aged 22-80 years, and 40 males aged 22-79 years). Vertebral heights were significantly higher in men than in women (p < 0.001). In women, a significant correlation was found between the height of T4 and the heights of the other vertebrae with correlation coefficients ranging from 0.71 (T5) to 0.38 (L1) and similar figures were found in males. Both the absolute and relative vertebral heights in this series were close to those reported by others. Selection of subjects and racial, environmental, and technical factors may explain the small differences. It is concluded that vertebral heights can be predicted from vertebrae rarely affected by spinal osteoporosis. Both relative and absolute vertebral heights differed between sexes. Although the reference values in this Scandinavian population were in agreement with previously reported data, small differences between populations were found, emphasizing the need for regional normative data. PMID- 8427750 TI - Normal age-related alterations on chest radiography. A longitudinal investigation. AB - To ascertain serial changes on chest radiography attributable to aging, 16 parameters and 4 pulmonary parenchymal manifestations were assessed at 2 intervals; namely, 18.5 and 10 years after initial radiography in healthy men (n = 51) and women (n = 149) (age range 41-82 years). The transverse cardiac diameter, cardiothoracic ratio, and aortic arch transverse diameter increased significantly in both sexes from the initial to the final examinations. Although several other factors changed significantly, these changes were relatively small and the individual variations large. Initially, 13% and finally, 27% of the subjects exhibited at least one of the selected pulmonary manifestations. PMID- 8427751 TI - Thrombus imaging with 99mTc-HMPAO-labeled platelets and 111In-labeled monoclonal antifibrin antibodies. AB - Eighteen patients with suspicion of deep venous thrombosis (DVT) in the lower extremities were imaged both with autologous 99mTc-HMPAO-labeled platelets (Tc PLT) and 111In-labeled monoclonal antifibrin antibodies (In-MoAbs) on the same day. Presence or absence of thrombosis was verified by venography. Tc-PLT was given i.v. followed after 30 min by In-MoAbs. Anterior and posterior projections of the lower extremities were obtained with a large field-of-view gamma camera at 5 to 25 min, 2 h, 4 to 6 h, and 20 h after administration of the marker. Both Tc PLT and In-MoAbs detected DVT well but less frequently than venography. Thrombi were visualized at 2 to 4 h after injection. The quality of images was better with Tc-PLT than with In-MoAbs. In the patients treated during the study, heparin significantly (p < 0.01) inhibited the uptake of Tc-PLT but not of In-MoAbs. We conclude that both Tc-PLT and In-MoAbs are suitable agents for the detection of DVT especially in patients without anticoagulation. PMID- 8427752 TI - Double-contrast enhanced MR imaging of myocardial infarction in the pig. AB - Myocardial infarction was induced by ligating a diagonal branch of the left anterior descending artery in 18 pigs. All pigs were sacrificed 6 h after the occlusion. Dysprosium diethylenetriamine-pentaacetic acid bismethylamide (Dy-DTPA BMA, 1.0 mmol/kg) was administered i.v. to 6 pigs, starting 3 min before sacrifice (injection time approximately 1 min). In a second group of 6 pigs, a double-contrast technique was used, consisting of an i.v. injection of gadolinium DTPA-BMA (0.4 mmol/kg) 2 h before sacrifice, followed by an i.v. injection of Dy DTPA-BMA (1.0 mmol/kg) 3 min before sacrifice. Six additional pigs, subjected to 6 h of coronary artery occlusion without administration of contrast medium, served as controls. The hearts were excised and imaged with MR. In the control animals, the infarctions demonstrated an increased signal intensity in the proton density- and T2-weighted images. Administration of Dy-DTPA-BMA primarily improved infarct visualization in the proton density- and T2-weighted images, due to reduction of signal intensity in nonischemic myocardium. The double-contrast technique further improved infarct visualization in all sequences. PMID- 8427753 TI - Iodixanol in cardioangiography in patients with coronary artery disease. Tolerability, cardiac and renal effects. AB - Iodixanol is a new nonionic, dimeric contrast medium. With the addition of 18 mmol/l Na+ and 0.3 mmol/l Ca++ to iodixanol 320 mg I/ml a plasma-isotonic solution was obtained. The purpose was to evaluate the suitability of iodixanol for use in cardioangiography by determining the diagnostic efficacy, patient tolerability, and cardiac and renal side-effects. Initially, 14 patients with coronary artery disease were examined using iodixanol. A double-blind, randomized study was then performed in 72 patients, comparing iodixanol and iohexol. Serum and urine were sampled before the examination, and one and 2 days after. The diagnostic information was good and the number of adverse events low with iodixanol. The patients reported significantly less of a sensation of warmth following injection of iodixanol than iohexol. Our results also indicate that iodixanol 320 mg I/ml influences renal function to a lesser degree than does iohexol 350 mg I/ml. We therefore conclude that isotonic iodixanol is a safe contrast medium for use in cardioangiography. PMID- 8427754 TI - Clot formation in angiographic catheters--an in vitro comparative study. Effects of heparin and protein coating of the catheter. AB - Clot formation was compared in 6 brands of angiographic catheters, each of which was tested in 3 states: untreated, heparinized, and albumin-coated. Forty-six samples of each brand in each state (in total 828) were filled with blood from 23 donors and incubated at +37 degrees C for 5 or 15 min. Physiologic saline was then injected through the catheters at a flow rate of 5 and 50 ml/s, respectively, and any expelled clots identified on a filter. Heparinization reduced the number of clotted catheters at 5 min, but not at 15 min. The only significant difference related to catheter brands was found between the heparinized nylon and polyurethane catheters at 5-min incubation with fewer clotted polyurethane catheters. This difference was not found between the same materials made by two other manufacturers. Albumin coating had no effect on clot formation, but clots were more easily expelled by low-flow injections in albumin coated and, to a lesser degree, in heparinized than in untreated catheters. PMID- 8427755 TI - Effect of intravenous contrast media on proximal and distal tubular hydrostatic pressure in the rat kidney. AB - The effect of i.v. injection of contrast media (CM, 1,600 mg I/kg b.w.) on proximal and distal tubular hydrostatic pressure (PTHP, DTHP) in the rat was investigated using a micropuncture technique. The PTHP and DTHP after injection of diatrizoate, iohexol, ioxaglate, or mannitol returned to control values within approximately 20 min. However, following iotrolan injection PTHP was still elevated above control levels after 35 min while DTHP remained elevated throughout the experiment (50 min). Iotrolan has a lower osmotic potential than the other CM when given in equivalent iodine doses. The concentration of iotrolan may thus increase more along the tubules than the other CM and consequently lead to a higher viscosity of urine, resulting in increases in PTHP and DTHP. The high intratubular pressure induced by iotrolan may explain our previous findings of reduced single nephron glomerular filtration rate caused by this CM. PMID- 8427756 TI - Effects of femoral arteriography and low osmolar contrast agents on renal function. AB - The contrast media-induced renal effects of 3 different low osmolar contrast agents were prospectively evaluated with creatinine, beta-2-microglobulin, and urea in serum before and after femoral arteriography in 110 consecutive patients. Forty-two patients (38%) had at least one of the 2 major risk factors for contrast media-induced nephropathy; diabetes mellitus or renal dysfunction. Six patients (5%) had both risk factors. There were statistically significant increases of S-creatinine (6.5%, p < 0.001) and S-beta-2-microglobulin (7.4%, p < 0.001) but no increase of S-urea, in the total patient material. This effect was independent of preexaminatory renal function, the presence of diabetes mellitus and type of low osmolar contrast agent used in these patients who were properly hydrated before the examination. The amount of administered contrast medium did not correlate with the degree of contrast agent-induced renal function impairment. PMID- 8427757 TI - Conversion factors between energy imparted to the patient and air collision kerma integrated over beam area in pediatric radiology. AB - Conversion factors between the energy imparted to the patient in pediatric radiography and air collision kerma integrated over beam area are presented. The values have been derived from Monte Carlo calculations in soft tissue phantoms and extend results published earlier to cover children from early infancy to the age of 15 years. Variations related to phantom size as well as to focus-phantom distance, radiation field size, orientation of view (a.p., lateral), tube potential, and beam filtration are given. We show that the conversion factor increases with increasing half-value layer of the X-ray beam and the anterioposterior width of the simulated child. Increasing the focus-phantom distance increases the conversion factor, while increasing the field size decreases the factors due to more scattered radiation escaping laterally from the phantom. PMID- 8427758 TI - Simple mathematical relations between the acetabular anteversion and sector angles. AB - Acetabular anteversion is an important parameter of the hip joint. It is measured at CT of the hip, as are the more recently introduced acetabular sector angles. These angles give information on the anterior and posterior support from the acetabulum to the femoral head. Some simple mathematical relationships between these angles are presented. PMID- 8427759 TI - Role of oxidised low density lipoprotein in atherogenesis. PMID- 8427761 TI - Atherosclerosis: what is it and why does it occur? PMID- 8427760 TI - Mechanisms of monocyte recruitment and accumulation. PMID- 8427762 TI - Smooth muscle cells and the pathogenesis of the lesions of atherosclerosis. AB - In this review we have tried to identify the characteristic features of SMCs in developing lesions of atherosclerosis and the extracellular factors that may be involved in regulating these altered features. Though the list seems long and complex there is probably a great deal of interplay among the different regulatory mechanisms. The function and activities of SMCs in the artery are dependent on the milieu created by the surrounding cells and the components of the extracellular matrix. In the normal, uninjured media of the artery, SMC phenotype and function seem to be in large part determined by the extracellular matrix in which they are embedded and by diffusible factors, in particular from endothelial cells. Endothelial cell injury, infiltration of monocytes and lymphocytes, and ultimately, thrombosis and platelet release, as seen in developing lesions of atherosclerosis, dramatically alter the balance of growth regulatory and vasoactive factors present in the local environment. These extracellular factors (table and figure) can alter both SMC phenotype, and thus responsiveness, and SMC migration, proliferation, and synthesis of the extracellular matrix. A better understanding of how specific factors mediate these responses, should make it possible to determine the ways in which the SMC response can be modulated. Though growth regulatory molecules seem to be key to this process, the challenge for the future is to understand their regulation in the environment of the artery wall and the interplay between growth-regulatory molecules, extracellular matrix, and vasoactive agents. PMID- 8427763 TI - Immune and inflammatory mechanisms in the development of atherosclerosis. PMID- 8427764 TI - Relation of coronary heart disease incidence and total mortality to plasma cholesterol reduction in randomised trials: use of meta-analysis. PMID- 8427765 TI - Arteriographic view of treatment to achieve regression of coronary atherosclerosis and to prevent plaque disruption and clinical cardiovascular events. AB - Lipid-lowering therapy, as assessed by angiography, clearly benefits the arterial disease process. For example, among intensively treated patients in FATS the frequency of definite progression per lesion at risk was reduced by 75% among mild and moderate lesions, which form the great preponderance of the lesion population. Regression frequency per lesion was more than doubled by intensive therapy in mild and moderate subgroups and quadrupled in the subgroup with severe lesions. Clinical events were reduced by 73%. This was clearly due to a 15-fold reduction in the likelihood that a mildly or moderately diseased arterial segment would undergo abrupt and substantial progression to a severe lesion at the time of the clinical event. It has been shown that the process of plaque fissuring, leading to plaque disruption, thrombosis, and clinical coronary events, is predicted by the size of the plaque core lipid pool and the abundance of lipid laden macrophages in its fibrous cap. Experimentally, lipid lowering therapy decreases the number of lipid-laden intimal macrophages and more slowly depletes core cholesteryl ester deposits. Thus the composite of new and previously published data presented here supports the idea that lipid-lowering therapy selectively lipid-depletes (causes regression of) those fatty lesions containing a large lipid core and abundant intimal foam cells. By doing so, these lesions, which are most vulnerable to fissuring, are rendered much more stable and the clinical event rate is accordingly decreased. PMID- 8427766 TI - Modification of atherosclerosis by agents that do not lower cholesterol. PMID- 8427767 TI - Cholesterol reduction in the prevention of coronary heart disease: therapeutic rationale and guidelines. The British Hyperlipidaemia Association. PMID- 8427768 TI - The fact and fiction of lowering cholesterol concentrations in the primary prevention of coronary heart disease. PMID- 8427769 TI - Blood flow modification by nicotinamide and metoclopramide in mouse tumours growing in different sites. AB - Nicotinamide (NA) and metoclopramide (MCA) have been shown to be sensitisers of the effects of radiation and drugs in experimental rodent tumours growing in skin and muscle. We have used 86Rb uptake to investigate the effects of these two drugs on the distribution of blood to a mouse carcinoma (NT) growing in skin, muscle or the gut wall, as well as to the host normal tissue. NA caused an increase in cardiac output distribution (COD) of between 17 and 92% to tumours in the three sites. When this increase is related to the changes in COD to the host normal tissues, however, COD to tumours in skin and muscle was increased by a factor of 1.8 and to tumours in the gut wall by a factor of 1.7. MCA caused a consistent increase in COD to tumours growing in muscle, but the effects in tumours in skin and gut were variable with time. Again when related to the change in COD to host normal tissues, a factor of 2.1 was seen for COD to tumours growing in muscle and gut. Both NA and MCA alter COD to tumours in some sites relative to host tissues in a way that could enhance anti-cancer drug delivery to tumours, though the effects of NA are more reliable in our systems. PMID- 8427770 TI - Micronucleus formation in human tumour cells: lack of correlation with radiosensitivity. AB - The micronucleus (MN) test has been carefully characterized in four human tumour cell lines of widely differing radiosensitivity. Two radioresistant bladder carcinoma cell lines (MGH-U1 and RT112), one sensitive medulloblastoma cell line (D283MED) and a sensitive neuroblastoma cell line (HX142) were used. The number of MN per Gy of ionising radiation was 0.13 for HX142, 0.17 for D283MED, 0.21 for RT112 and 0.26 for MGH-U1. This does not rank the cell lines in the same order of radiosensitivity as clonogenic cell survival where the surviving fraction at 2 Gy (SF2) was 0.11 for HX142, 0.2 for D283MED, 0.62 for RT112 and 0.53 for MGH-U1. This discrepancy between MN formation and cell death leaves doubt as to the potential usefulness of the MN test as a rapid assay of radiosensitivity but it has potential implications for the mechanistic basis of radiosensitivity in these cells. PMID- 8427771 TI - Proliferative behaviour of an oestrogen sensitive rat mammary tumour: evidence for a paracrine interaction between tumour and stroma. AB - An oestrogen-sensitive rat mammary tumour (OES HR1) has been grown in normal female rats and in female and male rats supplemented with oestrone. In some rats, after the tumour was established, both exogenous and endogenous sources of oestrogen were removed--a treatment which inhibited further growth of the tumour. The proliferative characteristics of the tumours were measured by injecting the rats with deoxybromouridine (BrdU) 4 h before removing the tumour. Extracted nuclei were reacted with anti-BrdU and the labelling index and DNA content measured by flow cytometry. A correlation between the number of (diploid) host cells present and the number of (aneuploid) tumour cells in S-phase of the cell cycle was observed. This result suggests that there are paracrine interactions between tumour and host cells. We also observed that, on oestrogen ablation, the labelling index was significantly reduced while the percentage of cells in S phase changed far less. The demonstration that there are cells in S-phase which are not proliferating highlights a possible problem with the measurement of proliferation in human tumours from a DNA histogram. PMID- 8427772 TI - Double megatherapy and autologous bone marrow transplantation for advanced neuroblastoma: the LMCE2 study. AB - In the LMCE1 study using a single course of megatherapy most of the relapses occurred during the first 2 years after autologous bone marrow transplantation. A second pilot study (LMCE2) was therefore set up using a double harvest/double graft approach with two different megatherapy regimens. Objectives were to test the role of increased dose intensity on response status, relapse pattern and overall survival. Thirty-three patients (20 boys, 13 girls) with a median age of 53 months at first megatherapy (range, 17-202 months) entered this study. They were cases either with refractory disease in partial response after second line treatment for stage 4 neuroblastoma (n = 25) or after relapse from stage 4 (n = 5) or stage 3 disease (n = 3). All patients received Etoposid and/or Cisplatinum (or Carboplatin) containing treatments before megatherapy. The first megatherapy regimen was a combination of Tenoposid, Carmustine and Cisplatinum (or Carboplatin), the second applied Vincristin, Melphalan and Total Body Irradiation. The first harvest was scheduled 4 weeks after the last chemotherapy, the second 60 to 90 days after megatherapy. All marrows were purged in vitro by an immunomagnetic technique. Median follow up time since first megatherapy is 56 months. Response rates for evaluable patients were 65% (complete response rate: 16%) for megatherapy 1 and 60% (complete response rate: 25%) for megatherapy 2. Considering that only patients with delayed response or relapse were eligible for this pilot study the overall survival was encouraging with 36% at 2 years and still 32% at 5 years. The costs for these survival rates were high in terms of morbidity (four early and four late toxic deaths; toxic death rate: 24%). Double harvesting may have the disadvantage of delayed engraftments related in part to a disturbance of marrow microenvironment by megatherapy 1. This double megatherapy approach achieved a prolonged relapse free interval (median 11 months, range 2-31 months) in patients reaching megatherapy 2 and justifies further evaluation of concepts with consecutive dose-escalation. PMID- 8427773 TI - DNA ploidy and S-phase in primary malignant melanoma as prognostic factors for stage III disease. AB - In 82 patients with stage III malignant melanoma, the primary tumours were investigated by DNA flow cytometry. The tumours were classified as DNA diploid (n = 36), tetraploid (n = 11) and aneuploid (n = 35). By univariate analysis a significant correlation with post-recurrence survival was found for time to first metastasis, DNA-ploidy and S-phase fraction. By multivariate analysis, significant prognostic variables were found to be the time to first metastasis (P = 0.006), and ploidy (P = 0.011). Patients with diploid melanomas and a long recurrence-free interval had a median post-recurrence survival time of 45 months compared to 18 months in patients with DNA aneuploid tumours and an early recurrence. The S-phase could be estimated in 47 primary melanomas and was found to be a significant prognostic variable (P = 0.017). The median survival was 45 months for patients with melanomas with a S-phase fraction below 5%, and 19 months for melanomas with S-phase above 10%. The prognostic value of the S-phase remained significant even after adjustment for recurrence-free interval and DNA ploidy. PMID- 8427774 TI - Treatment of recurrent and cystic malignant gliomas by a single intracavity injection of 131I monoclonal antibody: feasibility, pharmacokinetics and dosimetry. AB - A pilot study was undertaken to determine the feasibility of infusing 131I labelled monoclonal antibodies (MoAbs) into either the cavity remaining after resection of malignant glioma or into glioma cysts. Of the seven patients recruited into the study, two had cystic lesions and five resection cavities. Six of the seven were treated after relapse from primary therapy. All patients apart from one, were given a single injection of 131I conjugated to a MoAb (ERIC-1) recognising the human neural cell adhesion molecule (NCAM). One patient received a further injection of 131I-MoAb after regrowth of their disease. Pharmacokinetic studies revealed that the MoAb remained predominantly in the tumour cavity with little leakage into the systemic compartment. This resulted in a high calculated dose of radiation being delivered to the tumour cells either lining or within close proximity to the cavity/cyst wall. In such a small study, it is not possible to determine accurately response rates, but individual patient responses were observed. This, along with the low toxicity noted, demonstrates the feasibility of using 131I-MoAbs in this way. With 131I, radiation dose is deposited in tissue to a depth of 1 mm from the source. The possibility of applying isotopes such as 90Yttrium which will irradiate tumour/tissue to a greater depth (6 mm) is discussed in context with the biology of glioma infiltration into normal brain parenchyma. PMID- 8427776 TI - Cisplatin-associated anaemia treated with subcutaneous erythropoietin. A pilot study. AB - In 20 patients with cisplatin-associated anaemia (haemoglobin less than 90 gl-1), recombinant human erythropoietin was administered subcutaneously three times a week on an outpatient basis. The initial dose was 50 Units Kg-1 of body weight. If response was not achieved within 3 weeks, dose was increased to 75 Units Kg-1. Using the same criteria further escalation to 100 Units Kg-1 was performed. If there was no response erythropoietin was terminated. Fifteen patients obtained an increase in haemoglobin to above 100 gl-1 which was considered as a clinical response in this study, with a dose of 50 Units Kg-1; one patient needed an erythropoietin dose of 75 Units Kg-1 and one a dose of 100 Units Kg-1. Only three patients required haemotransfusions and were considered non responders. Haemoglobin increases occurred despite continuation of cisplatin chemotherapy. In conclusion subcutaneous low dose of erythropoietin seems to be effective and safe in the treatment of cisplatin-induced anaemia. PMID- 8427775 TI - Humoral mediation for cachexia in tumour-bearing rats. AB - Early and severe loss of body weight associated with pronounced tissue changes developed in rats transplanted with a fast-growing ascites hepatoma (Yoshida AH 130). The protein content showed an early and marked fall in the skeletal muscle, while in the liver it transiently increased 4 days after implantation then declined to values lower than in control animals. Protein loss in gastrocnemius muscle and liver resulted mainly from enhancement of protein catabolism (Tessitore L. et al., Biochem. J., 241: 153-158, 1987). In contrast to the tumour bearing rats, in the pair-fed animals the initial body weight was maintained, while the protein mass decreased sharply in the liver and moderately in the gastrocnemius muscle. In host animals total plasma protein decreased during the period of tumour growth, while both triglycerides and total cholesterol markedly increased. Glucose remained unchanged even when overt cachexia had developed. The total free amino acid concentration in the plasma of tumour-bearing rats decreased slightly by day 4 and returned to values close to those of controls in the late stages of tumour growth. By contrast, in the pair-fed controls the plasma levels of triglycerides and particularly of total free amino acids and glucose decreased over the whole experimental period, whereas total protein and cholesterol were unchanged. Marked perturbations in the hormonal homeostasis developed early after tumour transplantation. The plasma levels of glucagon, corticosterone and catecholamines rose sharply, while those of insulin and thyroid hormones decreased. Furthermore, high plasma concentrations of prostaglandin E2 (PGE2) and tumour necrosis factor (TNF) were observed over the whole experimental period. IL-1-like activity, TNF and PGE2 were released in vitro from AH-130 cells. These data suggest that the systemic effects of AH-130 tumour on the host rat reflected the interplay of a complex network of factors, including classical hormones and cytokines, all of which likely concur in enhancing tissue protein catabolism. PMID- 8427777 TI - Colorectal adenomas and energy intake, body size and physical activity: a case control study of subjects participating in the Nottingham faecal occult blood screening programme. AB - Most case-control studies of colorectal cancer have shown a positive association with energy intake. In contrast studies which have considered physical activity have found the most active to have a lower risk of colonic cancer and obesity appears to be no more than weakly related to colorectal cancer. We therefore compared energy intake determined by a diet history interview, self-reported height and weight, together with measures of lifetime job activity levels and leisure activity in the year prior to interview in 147 cases with colorectal adenomas and two control groups (a) 153 age-sex matched FOB-negative subjects (b) 176 FOB-positive subjects in whom no adenoma or carcinoma was found. Unconditional logistic regression was used to estimate relative risks (RR) and 95% confidence intervals () adjusted for age, sex and social class. No association with weight or body mass index was found. The only association with physical activity found with both control groups was an inverse association with running or cycling for half an hour continuously at least once a week RR 0.46 (0.2-1.3) compared with control group (a), and RR = 0.32 (0.1-0.8) compared with (b), but few subjects engaged in such activity. There was an inverse association with energy intake (trend chi 2 = 5.3, P < 0.025) in the comparison with control group (a) only, a finding which is consistent with those of two previous studies of asymptomatic adenoma. PMID- 8427778 TI - Hodgkin's disease diagnosed post mortem: a population based study. AB - All cases of Hodgkin's disease (HD) notified to the Danish Cancer Registry from 1976 through 1987 in patients less than 70 years old were reviewed in order to identify patients in whom a correct diagnosis was established only post mortem. The case records of such patients were reviewed in a search for clinical features that could have ensured a correct pre mortem diagnosis. HD was diagnosed after death in 31 patients in this unselected population based study and thus constituting only 2.4% of all patients less than 70 years with HD, but 14.1% of the group aged 65-69 years. Most patients were identified during the first part of the study period, which may reflect a decreasing autopsy rate. HD was considered to be a coincidental finding in four patients and the primary cause of death in 27 patients. Among the later 27 patients a number of unfavourable prognostic factors were a common finding: persistent unexplained fever and weight loss, pancytopenia, hepatic involvement, bone marrow involvement, advanced stage disease, and lymphocytic depletion histology. However, most of the patients had no concurrent diseases and may have benefitted from a correct diagnosis and a potentially curative treatment. The many uncommon features of HD together with the frequent findings of falsely negative chest X-ray, bone marrow examination, liver biopsy, and ultrasound contributed to the difficulty in diagnosis. In about 1/3 of the patients clinical findings suggestive of lymphoma did not result in relevant diagnostic procedures. PMID- 8427779 TI - Dose intensity analysis in advanced ovarian cancer patients. AB - To determine if chemotherapy dose intensity influences treatment outcome in advanced ovarian cancer, all randomised studies of first line chemotherapy, published between 1975 and 1989, were analysed for relationships between planned dose intensity and (a) objective response and (b) median survival. Total dose intensity of each study regimen was calculated and a weighted regression model providing for systemic differences in response or survival among studies was utilised. Hence, treatment arms of different studies were never directly compared. In addition, relative dose intensities of individual drugs within combinations was similarly evaluated. The improvement in objective response rate when adding one unit of total dose intensity ranged between 12% and 16% depending on baseline response rate. The improvement in median survival when adding one unit of total dose intensity ranged between 2 and 4 months. One unit of total dose intensity corresponds to, for example, 20 mg m2 week of cisplatin, or 25 mg m2 week of doxorubicin, or 350 mg m2 week of cyclophosphamide. The analysis of individual drugs suggested that doxorubicin and the platinum compounds were about equally effective, with cyclophosphamide being less effective. The methodological benefits and limitations of the approach used and the implication of the results are discussed. PMID- 8427780 TI - Acquisition of platinum drug resistance and platinum cross resistance patterns in a panel of human ovarian carcinoma xenografts. AB - In vivo models of acquired resistance to the platinum-based agents cisplatin (CDDP), carboplatin (CBDCA), iproplatin (CHIP) and tetraplatin have been established using a panel of six parent human ovarian carcinoma lines, two (HX/110 and PXN/87) being derived from previously untreated patients. Resistance has been generated to CDDP (three lines), CBDCA (one line), CHIP (three lines) and tetraplatin (one line) either by treatment in vivo or (for one line to CDDP) through exposure in vitro and subsequent transfer to mice. With the four tumours where resistance was generated using CDDP or CBDCA, a complete cross-resistance to the remaining platinum agents studied was observed. In contrast, in one of three lines with derived resistance to the platinum (IV) agent, CHIP, (PXN/951) a retention in sensitivity was observed with CDDP and CBDCA. Only one of the six parent tumour lines (PXN/100) was markedly sensitive to tetraplatin. Where resistance was generated to tetraplatin (PXN/100T) there was some retention of activity by CDDP. For the CDDP-resistant line established in vitro, there was a close agreement between the cross-resistance profile obtained in vitro vs that obtained in vivo. This tumour panel may be useful in the elucidation of cellular and molecular resistance mechanisms to platinum drugs operative in vivo. Moreover, as they appear to mimic the clinical observations of shared cross resistance between CDDP, CBDCA and CHIP, they may represent valuable preclinical evaluation models for the discovery of drugs capable of conferring responses in CDDP-refractory ovarian cancer. PMID- 8427781 TI - Deoxyribonuclease treatment prevents blood-borne liver metastasis of cutaneously transplanted tumour cells in mice. AB - Murine L5178Y-ML cells, when transplanted subcutaneously into the flank of (BALB/c x DBA/2)F1 mice, grew locally and always formed spontaneous metastases in the liver. Even after surgical removal of the primary tumour mass 5 or 7 days after tumour cell inoculation, all mice died due to liver metastases within 18 days. Using this model of tumour metastasis, we examined whether serine protease or deoxyribonuclease I (DNase I) would affect metastasis. Spontaneous liver metastasis of L5178Y-ML cells was enhanced by systemic administration of alpha chymotrypsin at 3, 4 and 5 days or at 5, 6 and 7 days after tumour cell inoculation. This result was consistent with a previous report on blood-borne lung metastasis. In contrast, systemic administration of DNase I at 3, 4 and 5 days or at 5, 6 and 7 days after tumour cell inoculation inhibited liver metastasis. Neither treatment affected primary tumour growth. An influence of DNase I on tumour cell arrest in the microvasculature of the liver was suggested by scanning electron microscopy. DNase I treatment resulted in a statistically significant prolongation of the survival period, however, the effect was not satisfactory. A more striking anti-metastatic treatment resulting in a greater prolongation of the survival period was achieved by combining surgical removal of the primary tumour mass with DNase I treatment. These results suggest that DNase I could be a potential therapeutic agent used in conjunction with surgery to prevent clinical blood-borne metastasis. PMID- 8427782 TI - Epidermal growth factor receptor in breast cancer. Comparison with non malignant breast tissue. AB - Epidermal growth factor receptors were measured using a radioligand binding assay in membrane preparations from 67 cancer and 25 non-malignant tissues. The binding characteristics of EGFr were similar in tumour and normal breast membranes. The concentrations were significantly higher in non-malignant tissue than in cancer. EGFr concentrations were directly correlated with steroid receptors in non malignant tissue, whereas in cancer an inverse correlation between EGFr and steroid receptors was found. PMID- 8427783 TI - pH dependent cytotoxicity of N-dodecylimidazole: a compound that acquires detergent properties under acidic conditions. AB - N-dodecylimidazole is a compound which acquires detergent properties under acidic conditions and might be useful in killing selectively cells in those regions of solid tumours which have a reduced extracellular pH (pHe). We have therefore studied the effects of N-dodecylimidazole against malignant cells in tissue culture. N-dodecylimidazole displayed pHe-dependent cytotoxicity against EMT-6 and MGH U1 cells; cell killing was dose dependent and was 100-fold greater at pHe 6.0 than pHe 7.0. Reduced toxicity of N-dodecylimidazole was observed at higher cell concentrations (> 10(6) cells ml-1), and only minor effects were observed against multicellular tumour spheroids. Potential mechanisms of action of N dodecylimidazole include detergent-mediated lysis of the cell membrane at low pHe, and selective uptake into lysosomes where detergent activity leads to rupture of the lysosomal membrane and release of cytolytic enzymes. Inhibition of activity of cysteine proteases by the inhibitor E-64 did not protect cells against the toxicity of N-dodecylimidazole, suggesting that these lysosomal enzymes do not play a major role in the mechanism of action of this compound. Lysis of erythrocytes (which contain no lysosomes) was observed with low concentrations of N-dodecylimidazole. Dependence of cell lysis on cell concentration was similar to that observed for two other detergents that act on the plasma membrane, Triton X-100 and sodium dodecyl sulfate. We conclude that N dodecylimidazole causes pHe dependent cell killing in two cultured tumour cell lines, and that its mechanism of action is probably due to acid mediated production of detergent activity which acts primarily on the cell plasma membrane. PMID- 8427784 TI - The TP53 tumour suppressor gene in colorectal carcinomas. II. Relation to DNA ploidy pattern and clinicopathological variables. AB - Heterozygous loss of the TP53 gene on chromosome arm 17p in colorectal carcinomas was strongly associated with DNA aneuploidy (P < 0.0001). This association was seen only in tumours with loss on both 17p and 17q (P < 0.001), but not for loss on 17p only. DNA near diploid (ND) carcinomas and DNA aneuploid (AN) tumours with DNA index > or = 1.1 and < 1.3 had similar frequencies of TP53 gene loss (49% and 42%, respectively), whereas AN tumours with DNA index > or = 1.3 had a significantly higher frequency of TP53 gene loss (85%) (P < 0.0001 and P < 0.0001, respectively). There was a significant association between loss of the TP53 gene and histological grade (P < 0.01), and there tended to be an association between loss of the TP53 gene and degree of cellular atypia (P < 0.05), with TP53 gene loss being most frequent in moderately differentiated carcinomas, and in carcinomas with severe cellular atypia, respectively. The proportion of tumours with loss of the TP53 gene increased significantly towards the distal part of the large bowel (P < 0.0001). These results indicate that different genetic mechanisms may be involved in the carcinogenesis in colon and rectum carcinomas, and in the two subsets of DNA aneuploid carcinomas. Furthermore, the data may suggest a role for the TP53 gene in the aneuploidisation process, possibly as a 'target' for a whole chromosome loss. PMID- 8427785 TI - Dexamethasone decreases urokinase plasminogen activator mRNA stability in MAT 13762 rat mammary carcinoma cells. AB - The glucocorticoid dexamethasone was observed to decrease urokinase plasminogen activator (uPA) RNA levels from within 1 h of treatment of MAT 13762 mammary adenocarcinoma cells. The drug did not alter the rate of uPA gene transcription in these cells, but decreased the stability of cytoplasmic uPA mRNA transcripts. Results from cycloheximide and actinomycin D experiments indicated that the dexamethasone-mediated reduction in uPA RNA required both new protein and RNA synthesis. Based on these results, we propose that dexamethasone induces a short lived protein(s) which down-regulates uPA RNA levels post-transcriptionally in these metastatic tumour cells. PMID- 8427786 TI - Nurses praised for 'major contribution' to AIDS care. PMID- 8427787 TI - Cheaper, more accurate TB test is in the pipeline. PMID- 8427788 TI - Immunoglobulins offer hope in AIDS survival. PMID- 8427789 TI - Romania: something to celebrate. Interview by Norah Casey. PMID- 8427790 TI - Christmas quiz: assess your stress! PMID- 8427791 TI - Nursing abroad: nursing in Nepal. PMID- 8427792 TI - Infertility: causes and treatment. AB - Issues related to infertility appear with increasing regularity in the media. Many people who suffer from this problem are surprised and shocked to find themselves attending an infertility clinic. The author describes the most common types of infertility and explores the treatments available. In particular, she examines male infertility and the impact this has on the couple. PMID- 8427793 TI - The A-J of trauma assessment in A&E. AB - A thorough initial assessment of trauma patients can be life saving. As nurses are usually the first people to receive a patient in accident units, the author describes a method which can help nurses to assess the patient's condition systematically. PMID- 8427794 TI - Elderly, homeless and mentally ill: a study. AB - This article describes a study of people aged 60 and over who were homeless and sleeping on the streets in Central London. The author spent 334 hours on the streets of London between July and October 1990, and attempted interviews with 130 elderly homeless people before finding 50 who were able to give a full interview. Information about this client group was then collected, including mental health problems they reported or that were observed during the interviews. The results suggest that the link between mental health problems and homelessness among elderly people may be profound. PMID- 8427795 TI - A Christmas story. Sitting next to Nellie. PMID- 8427797 TI - Christmas presents. Faint scent of giving is bottom of the list. PMID- 8427796 TI - The Clay column. PMID- 8427798 TI - Elderly care: give Santa the sack. PMID- 8427799 TI - Holiday cheer: a 'hiccup' in Parliament. PMID- 8427800 TI - Classification of Rhizomonas suberifaciens, an unnamed Rhizomonas species, and Sphingomonas spp. in rRNA superfamily IV. AB - Thermal melting profiles of hybrids between 3H-labeled rRNA of Rhizomonas suberifaciens, the causal agent of corky root of lettuce, and chromosomal DNAs from 27 species of gram-negative bacteria indicated that the genus Rhizomonas belongs to superfamily IV of De Ley. On the basis of the melting temperatures of DNA hybrids with rRNAs from the type strains of R. suberifaciens, Sphingomonas paucimobilis, and Sphingomonas capsulata, Rhizomonas strains constitute a separate branch in superfamily IV, which is closely related to but separate from branches containing Zymomonas mobilis, Sphingomonas spp., and S. capsulata. Sphingomonas yanoikuyae and Rhizomonas sp. strain WI4 are located toward the base of the Rhizomonas rRNA branch. DNA-DNA hybridization indicated that S. yanoikuyae is equidistant from Rhizomonas sp. strain WI4 and S. paucimobilis. Sequences of 270 bp of 16S ribosomal DNAs from eight strains of Rhizomonas spp., eight strains of Sphingomonas spp., and Agrobacterium tumefaciens indicated that S. yanoikuyae and Rhizomonas sp. strains WI4 and CA16 are genetically more closely related to R. suberifaciens than to Sphingomonas spp. Thus, S. yanoikuyae may need to be transferred to the genus Rhizomonas on the basis of the results of further study. PMID- 8427801 TI - Phylogeny of the ammonia-producing ruminal bacteria Peptostreptococcus anaerobius, Clostridium sticklandii, and Clostridium aminophilum sp. nov. AB - In previous studies, gram-positive bacteria which grew rapidly with peptides or an amino acid as the sole energy source were isolated from bovine rumina. Three isolates, strains C, FT (T = type strain), and SR, were considered to be ecologically important since they produced up to 20-fold more ammonia than other ammonia-producing ruminal bacteria. On the basis of phenotypic criteria, the taxonomic position of these new isolates was uncertain. In this study, the 16S rRNA sequences of these isolates and related bacteria were determined to establish the phylogenetic positions of the organisms. The sequences of strains C, FT, and SR and reference strains of Peptostreptococcus anaerobius, Clostridium sticklandii, Clostridium coccoides, Clostridium aminovalericum, Acetomaculum ruminis, Clostridium leptum, Clostridium lituseburense, Clostridium acidiurici, and Clostridium barkeri were determined by using a modified Sanger dideoxy chain termination method. Strain C, a large coccus purported to belong to the genus Peptostreptococcus, was closely related to P. anaerobius, with a level of sequence similarity of 99.6%. Strain SR, a heat-resistant, short, rod-shaped organism, was closely related to C. sticklandii, with a level of sequence similarity of 99.9%. However, strain FT, a heat-resistant, pleomorphic, rod shaped organism, was only distantly related to some clostridial species and P. anaerobius. On the basis of the sequence data, it was clear that strain FT warranted designation as a separate species. The closest known relative of strain FT was C. coccoides (level of similarity, only 90.6%). Additional strains that are phenotypically similar to strain FT were isolated in this study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427802 TI - Differentiation of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of cell proteins. AB - The protein patterns of whole cells of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillarum were studied by using sodium dodecyl sulfate polyacrylamide gel electrophoresis. The protein patterns of the 16 strains of E. rhusiopathiae and E. tonsillarum studied, including the type strains of these two species, resembled each other, except that there were 71-, 41-, 34-, and 26-kDa proteins in the E. rhusiopathiae pattern and 74-, 44-, 36-, and 25-kDa proteins in the E. tonsillarum pattern. This observation indicates that there is some phenotypic heterogeneity in the genus Erysipelothrix. In addition, the protein patterns of E. rhusiopathiae serotype reference strains representing serotypes 1 through 23 and type N were compared. The protein patterns of serotype 1a, 1b, 2, 4, 5, 6, 8, 9, 11, 12, 15, 16, 19, and 21 and type N strains were similar to the pattern of the type strain of E. rhusiopathiae (strain ATCC 19414). Conversely, the protein patterns of serotype 3, 7, 10, 14, and 20 strains were very similar to the pattern of the type strain of E. tonsillarum (strain ATCC 43339). An atypical pattern was observed in serotype 13, 17, 18, 22, and 23 strains. These results suggest that this method may be used as an aid in studying the taxonomy of these bacteria. PMID- 8427803 TI - Telluria mixta (Pseudomonas mixta Bowman, Sly, and Hayward 1988) gen. nov., comb. nov., and Telluria chitinolytica sp. nov., soil-dwelling organisms which actively degrade polysaccharides. AB - Pseudomonas mixta (type strain, ACM 1762 [= ATCC 49108], an actively dextranolytic species that possesses both lateral and polar flagella, was compared with the strictly aerobic, rod-shaped, chitinolytic bacterium "Pseudomonas chitinolytica" ACM 3522T (= CNCM I-804) (T = type strain), which has a similar flagellation pattern, by performing phenotypic characterization and DNA DNA hybridization studies and by analyzing DNA base compositions and 16S rRNA sequences. Our results indicated that "P. chitinolytica" ACM 3522T was phenotypically and genotypically distinct from P. mixta and other phenotypically analogous Pseudomonas spp., Xanthomonas maltophilia, and other aerobic chitin degraders. The 16S rRNA sequences of strains ACM 1762T and ACM 3522T were found to be very similar (97%) to each other and indicated that these organisms are proteobacteria that belong to the beta subclass. The strains were deeply branched in the beta subclass and were distinct from other pseudomonads, including Pseudomonas cepacia, and from Comamonas testosteroni. On the basis of phenotypic, genotypic, and phylogenetic evidence, it is proposed that P. mixta and "P. chitinolytica" ACM 3522T represent two distinct species in a new genus called Telluria. Thus, the genus Telluria gen. nov. contains Telluria mixta comb. nov. and Telluria chitinolytica sp. nov., which are strictly aerobic, rod-shaped, soil dwelling bacteria that are active polysaccharide degraders. PMID- 8427804 TI - Porphyrobacter neustonensis gen. nov., sp. nov., an aerobic bacteriochlorophyll synthesizing budding bacterium from fresh water. AB - Four strains of orange- or red-pigmented bacteria isolated from freshwater surfaces were shown to synthesize bacteriochlorophyll under aerobic conditions. These strains shared unusual morphological features, such as acellular stalks, crateriformlike structures, and buds, with bacteria in the order Planctomycetales. However, comparisons of 16S rRNA sequences showed them to be members of the alpha-4 subdivision of the class Proteobacteria and most closely related to the marine aerobic bacteriochlorophyll-synthesizing bacterium Erythrobacter longus. They also differ from members of the Planctomycetales phenotypically in their synthesis of bacteriochlorophyll and possession of a peptidoglycan cell wall. They can be distinguished from E. longus on the basis of their 16S rRNA sequence, the G+C content of their DNA, cellular fatty acid composition, and carbon substrate spectrum. A new genus, Porphyrobacter, with a single species, P. neustonensis gen. nov., sp. nov., is proposed for these strains. The type strain is ACM 2844. PMID- 8427805 TI - Thauera selenatis gen. nov., sp. nov., a member of the beta subclass of Proteobacteria with a novel type of anaerobic respiration. AB - A recently isolated, selenate-respiring microorganism (strain AXT [T = type strain]) was classified by using a polyphasic approach in which both genotypic and phenotypic characteristics were determined. Strain AXT is a motile, gram negative, rod-shaped organism with a single polar flagellum. On the basis of phenotypic characteristics, this organism can be classified as a Pseudomonas sp. However, a comparison of the 16S rRNA sequence of strain AXT with the sequences of other organisms indicated that strain AXT is most similar to members of the beta subclass (level of similarity, 86.8%) rather than to members of the gamma subclass (level of similarity, 80.2%) of the Proteobacteria. The presence of the specific polyamine 2-hydroxyputrescine and the presence of a ubiquinone with eight isoprenoid units in the side chain (ubiquinone Q-8) excluded strain AXT from the authentic genus Pseudomonas and allowed placement in the beta subclass of the Proteobacteria. Within the beta subclass, strain AXT is related to Iodobacter fluvatile. The phylogenetic distance (level of similarity, less than 90%), as well as a lack of common phenotypic characteristics between these organisms, prevents classification of strain AXT as a member of the genus Iodobacter. In addition, strain AXT possesses a unique mechanism for anaerobic respiration, which allows it to utilize selenate as an electron acceptor without interference by nitrate. Therefore, we propose that strain AXT should be the first member of a new genus and species, Thauera selenatis. PMID- 8427806 TI - A new leptospiral serovar in the Icterohaemorrhagiae serogroup isolated from an ox in Zimbabwe. AB - A strain of Leptospira interrogans that was isolated from an ox slaughtered in Zimbabwe and belonged to serogroup Icterohaemorrhagiae could not be identified when we compared it with 18 reference strains belonging to this serogroup by using cross-agglutinin absorption, monoclonal antibody, and restriction endonuclease DNA analyses. The name zimbabwe is proposed for the new serovar containing this strain; the type strain of this serovar is strain SBF 23. PMID- 8427807 TI - Optical DNA-DNA homology in the genus Listeria. AB - Twenty-three strains of the seven recognized Listeria species were studied by using DNA-DNA optical hybridization. The level of error in the data was low. Our results supported the results of Rocourt et al. (J. Rocourt, F. Grimont, P. A. D. Grimont, and H. P. R. Seeliger, Curr. Microbiol. 7:383-388, 1982), although there was some overlap between Listeria monocytogenes and Listeria innocua. We suggest that there may be more than one cluster in the species L. monocytogenes or the species may form a large spectrum of relatedness. The level of intraspecies homology in L. monocytogenes is very broad, as determined in both this study and other studies. PMID- 8427808 TI - Mycoplasma indiense sp. nov., isolated from the throats of nonhuman primates. AB - Mycoplasmas isolated from the throats of a rhesus monkey and a baboon within 3 days of their arrival from India were shown to be serologically distinct from 104 previously recognized Mycoplasma and Acholeplasma spp. Two mycoplasma colonies were cloned and examined in detail for morphology, growth, and biochemical characteristics. The two strains were closely related and had the following properties: guanine-plus-cytosine content of 32 mol%, requirement for sterol, arginine hydrolysis, and anaerobic growth. Glucose was not metabolized, and urea was not hydrolyzed. Strain 3T (= NCTC 11728) is the type strain of a new species, Mycoplasma indiense. PMID- 8427809 TI - A taxonomic review of the genus Microbispora by analysis of ribosomal protein AT L30. AB - We analyzed the ribosomal AT-L30 proteins from 13 type strains of species belonging to the genera Microbispora and Actinomadura. The electrophoretic mobilities of the AT-L30 preparations from Microbispora strains, as determined by two-dimensional polyacrylamide gel electrophoresis, revealed that the members of the genus Microbispora are phylogenetically homogeneous. The results of partial amino acid sequencing of AT-L30 preparations from several representative Microbispora strains supported the separation of the genus Microbispora from other related genera. The amino acid sequences of the AT-L30 proteins from strains of species belonging to the genus Actinomadura sensu stricto displayed a diversity that exemplified the low levels of amino acid sequence homology within the genus. This diversity was considered to be a characteristic typical of the genus Actinomadura. PMID- 8427810 TI - A new approach to use of bacteriolytic enzymes as a tool for species identification: selection of species-specific indicator strains with bacteriolytic activity towards Enterococcus strains. AB - We describe the bacteriolytic activity of 377 group D Enterococcus isolates expressed towards 25 Enterococcus strains belonging to different species and Micrococcus luteus ATCC 4698. Of the 26 indicator strains used to reveal bacteriolytic activity, 5 were lysed by all of the strains of some species and were not lysed by all of the strains of other species. The use of these indicator strains allowed us to devise a new method to differentiate group D Enterococcus strains, based on qualitative analysis (lysis or no lysis of the indicator strains) of bacteriolytic activity. The bacteriolytic patterns obtained fell into six bacteriolytic groups corresponding (98% agreement) to species or groups of enterococci as determined by a comparison with data from a phenetic similarity study. PMID- 8427811 TI - Phylogenetic relationships of marine bacteria, mainly members of the family Vibrionaceae, determined on the basis of 16S rRNA sequences. AB - The phylogenetic relationships of 50 reference strains, mostly marine bacteria which require Na+ for growth, were determined on the basis of 600 16S rRNA nucleotides by using reverse transcriptase sequencing. Strains belonging to 10 genera were included (four genera of the family Vibrionaceae, the genus Aeromonas of the family Aeromonadaceae, and the genera Alteromonas, Marinomonas, Shewanella, Pseudomonas, and Deleya). The sequences were aligned, the similarity values and evolutionary distance values were determined, and a phylogenetic tree was constructed by using the neighbor-joining method. On the basis of our results, the family Vibrionaceae was separated into at least seven groups (genera and families). Vibrio marinus clearly was on a line of descent that was remote from other vibrios. As determined by the similarity and evolutionary distance values, V. marinus is more distantly related to the family Vibrionaceae than the members of the Aeromonadaceae are. Also, Vibrio cholerae strains formed a separate group with Vibrio mimicus at the genus level. Of 30 species of the Vibrionaceae, 17 formed a large phylogenetic cluster. The genus Listonella was found to be a heterogeneous group, and the species were distributed in various subgroups of the Vibrionaceae. The separation of the family Aeromonadaceae from the family Vibrionaceae and the separation of the genera Marinomonas and Shewanella from the genus Alteromonas were confirmed in this phylogenetic study. However, a marine Pseudomonas species, Pseudomonas nautica, was clearly separated from two terrestrial Pseudomonas species. Each group that was separated by the phylogenetic analysis had characteristic 16S rRNA sequence patterns that were common only to species in that group. Therefore, the characteristic sequences described in this paper may be useful for identification purposes. PMID- 8427812 TI - Structural relationship between epidermal lipid lamellae, lamellar bodies and desmosomes in human epidermis: an ultrastructural study. AB - The water permeability of the stratum corneum (SC) appears to be regulated primarily by the lamellar arrangement of lipid bilayers between the corneocytes. A significant body of evidence already exists, suggesting that the specific structural organization of these intercellular lipid lamellae is responsible for the very low water permeability of the intact skin and that these lipid-rich structures may also influence the process of desquamation in the SC. In this electron microscopic study the structure of the intercellular domains at different levels within the SC of normal skin from 18 healthy subjects has been evaluated with a special fixation protocol utilizing acrolein vapour as primary fixation, followed by a modified ruthenium tetroxide (RuO4)-post-fixation technique. This procedure permitted an insight into the process of post-secretory extracellular processing of the lamellar body (LB)-derived lipids into lamellar lipid bilayers. This transformation takes place in unique saccular invaginations of the intercellular domains, which indent the underlying stratum granulosum (SG) cells. In this specialized environment LB lipids are first processed into broad sheets before they become part of the typical lamellar lipid structure of the SC. Furthermore, in the process of lipid maturation distinct differences between inner and outer parts of the SC emerge, in particular an increase in both the number of the lamellae per intercellular space, and their order of arrangement. Moreover, distinct structural relationships between desmosomes (at the SG/SC interface and lower SC) and desmosomal remnants (at the stratum disjunctum) on the one hand, and lipid layers on the other, have been demonstrated, pointing to an important functional interaction of these components in normal human skin. PMID- 8427813 TI - Are atopic disorders due to a TH2 clone? PMID- 8427814 TI - Lupus erythematosus profundus treated with clobetasol propionate under a hydrocolloid dressing. PMID- 8427815 TI - Colocalization of lichen planus and vitiligo. PMID- 8427816 TI - Keratodermia palmo-plantaris papulosa (Buschke-Fischer's disease): efficacy of acitretin. PMID- 8427817 TI - Nephrotoxicity due to azathioprine. PMID- 8427818 TI - Dynamic expression of pemphigus and desmosomal antigens by cultured keratinocytes. AB - Dynamic expression of pemphigus antigens by cultured human and mouse keratinocytes was compared with that of desmosome-associated molecules and cellular markers relating to epidermal differentiation. Plakoglobin was detected in localized areas of keratinocyte sheets in low Ca2+ (0.15 mM) KGM medium. In minimum essential medium (MEM) containing 1.8 mM Ca2+, plakoglobin was expressed in the intercellular spaces (ICS) throughout the keratinocyte sheet. Desmoplakin I and II, which were present in the cytoplasm of keratinocytes in the low Ca2+ medium, moved to the cell surface after the medium was changed to MEM. Desmoglein 1 and pemphigus vulgaris (PV) antigens were observed in the ICS of both the monolayers and stratified areas in the MEM. Pemphigus foliaceus (PF) antigens, frequently together with desmoglein 1, involucrin and keratins specific for the upper layer of the epidermis, were expressed by stratified keratinocytes but not the cells in the monolayers. The Western blotting study of the cultured keratinocyte extract showed 160- and 130-kDa bands positive for desmoglein 1 antigens and a 130-kDa band stained with PV sera. These findings suggest that although desmoglein 1 molecules bear PF antigenic sites, their expression pattern by cultured keratinocytes is closely related to that of PV rather than PF antigens. The PF antigenic sites may be formed on desmoglein 1 during epidermal differentiation. PMID- 8427819 TI - In vitro lymphocyte reactivity to heavy metal salts in the diagnosis of oral mucosal hypersensitivity to amalgam restorations. AB - The value of lymphocyte reactivity in the diagnosis of hypersensitivity to amalgam restorations was studied in a group of patients with oral mucosal lesions, and in a control group. Heavy metal salts were added to lymphocyte cultures, and lymphocyte proliferation and levels of IL-2 receptors and interferon-gamma (IFN-gamma) were determined. After addition of mercuric chloride, a statistically significant increase of IFN-gamma was seen in the patient group, indicating a higher reactivity to this metal salt. Mercuric chloride, nickel sulphate, palladium chloride and zinc chloride all stimulated lymphocyte proliferation, but there was no difference between patients and controls. Nine other metal salts tested did not influence the proliferation rate in either group. Mercuric chloride, nickel sulphate, and zinc chloride did not have any effect on the level of IL-2 receptors. Thus, certain functional tests other than crude lymphocyte proliferation may be useful in the diagnosis of amalgam hypersensitivity. PMID- 8427820 TI - Effect of etretinate on cyclosporin metabolism in vitro. AB - Cyclosporin (CyA) is an effective treatment for psoriasis, including cases unresponsive to other therapies. The major side-effect of CyA treatment is dose related nephrotoxicity. Combinations of CyA and etretinate (Et) have been tested with a view to reducing CyA dose requirements, and therefore minimizing adverse effects. We have studied the effect of Et on the cytochrome P-450-mediated metabolism of CyA. Microsomes prepared from histologically normal human (obtained from four cadaver kidney transplant donors; all male; age range 21-56) were incubated with CyA and various concentrations of Et. Metabolism was quantified by high-performance liquid chromatography with radiometric detection, and metabolites tentatively identified from the retention times of authentic standards. After 30 min incubation of CyA and microsomal protein at 37 degrees C, 10.1 +/- 3.0% (mean +/- SD) 3H-CyA was converted to the monohydroxylated metabolites M1 and M17, and 3.3 +/- 0.8% to the N-demethylated metabolite M21. At an Et concentration of 100 microM inhibition of CyA hydroxylase and N-demethylase was < 20%. This study indicates that there is no metabolic interaction between CyA and Et in vitro; it is likely that the two drugs are metabolized by different P-450 isoenzymes. PMID- 8427821 TI - Topical betamethasone-17-valerate inhibits heat-induced vasodilatation in man. AB - Topical betamethasone-17-valerate inhibits the vasodilatation response to local non-pathological heating of rat skin. We have shown that this effect can also be demonstrated in man. Topical betamethasone-17-valerate significantly inhibited the increase in human forearm cutaneous blood flow in response to heat of 44 degrees C, as measured by laser-Doppler velocimetry. This suggests that the effects of topical steroids upon skin blood flow in the rat and in man are similar, and supports the use of the animal model as a paradigm for studying human skin blood flow, and changes in response to anti-inflammatory steroids. PMID- 8427822 TI - Narrow-band (TL-01) UVB air-conditioned phototherapy for chronic severe adult atopic dermatitis. AB - In an open study of 21 severely affected adult atopic dermatitis patients, air conditioned narrow-band UVB phototherapy using the Philips TL-01 lamp three times weekly for 12 weeks resulted in a 68% reduction in atopic dermatitis severity scores, with a concomitant 88% reduction in potent topical steroid use. Follow-up at 24 weeks revealed that six patients had relapsed to > 70% of pre-phototherapy severity scores; the remaining 15 continued to derive long-term benefit. The mean value of potent topical steroid use remained 50% below pre-phototherapy needs. Narrow-band UVB (TL-01) phototherapy appears an effective, steroid-sparing treatment for chronic severe atopic dermatitis, offering long-term benefits in the majority of those treated. PMID- 8427823 TI - Nail varnish allergy with far-reaching consequences. AB - Contact allergy to nail varnish is well-known, and toluene-sulphonamide formaldehyde resin (TSAfr) was identified as the main allergen in 1943. During the period October 1989-December 1991 we identified 18 cases of contact allergy to nail varnish. The aim of the study was to describe the clinical picture, patch test results, course and socio-medical consequences. Seventeen of the 18 patients were patch-test positive to TSAfr and 17/18 were positive to their own nail varnishes. Fourteen of the 18 were also positive to one or more substances in the standard patch-test series. The lesions were scattered, involving the face, eyelids, neck and hands. Periungual lesions were recorded in 11/18. The dermatitis resolved within a few weeks when the use of nail varnish was stopped. The socio-medical consequences of contact allergy to nail varnish had been severe: sick leave (nine cases), hospitalization (four cases), cessation of visual-display-unit (VDU) work (two cases), and job-loss (two cases). Our conclusions are that contact allergy to nail varnish and TSAfr is common; the socio-medical consequences may be severe; periungual lesions occur more frequently than previously stated, and the presence of other contact allergies makes the diagnosis easy to miss. TSAfr should be included in the standard patch test series and patients should also be tested with their own nail varnishes. The study illustrates the need for mandatory declaration of the ingredients of cosmetics, as is required in the U.S.A. PMID- 8427824 TI - Assessment of area of involvement in skin disease: a study using schematic figure outlines. AB - The ability to assess the severity of dermatoses by measurement of area of involvement is important in both clinical practice and research. Using schematic figure outlines we have shown that physicians, nurses and other groups are unable to assess area accurately, and that the degree of error is partially dependent on absolute area, but also on other factors. The pattern and magnitude of the errors varied between the groups examined, but overall was such that it calls in question the use of area indices such as the PASI (psoriasis area and severity index) score in the assessment of inflammatory dermatoses. PMID- 8427825 TI - The suitability of SunCheck patches and Tanscan cards for monitoring the sunburning effectiveness of sunlight. AB - UV-sensitive products which undergo a colour change on exposure to UV radiation are available for use by the general public. We have evaluated SunCheck patches of various sensitivities, and Tanscan UV sensor cards, in terms of their temperature stability, wavelength response and response to sunlight. The products exhibited no perceptible colour change when the temperature was maintained at 45 degrees C for 4 hours; on exposure to monochromatic radiation, the sensitivity peaked at a wavelength of 335 and 355 nm for SunCheck and Tanscan, respectively; the response to sunlight under clear conditions was reasonably consistent, with the coefficient of variation for each type of material ranging between 12.5 and 27%. These products are based on the incorrect concept that there is a 'safe' dose of UV radiation. Thus, they cannot be recommended to the public as a reliable way of reducing the risk of skin cancer. PMID- 8427826 TI - Epidermolysis bullosa simplex, Dowling-Meara type. A report of two cases with different types of tonofilament clumping. AB - Two cases of the Dowling-Meara type of epidermolysis bullosa simplex (EBS) are described. Both had severe blistering at birth, which improved gradually with age. Vesicles and small bullae clustering in a herpetiform fashion were seen in both cases. One showed mild pincer deformity of the nails, and in the other the nail plates were shed after subungual blistering, but regrew without deformity. Histopathology and ultrastructural study showed cytolysis of the basal cells in both cases, but ultrastructurally different forms of tonofilament clumps were present in epidermal keratinocytes. In one case there was typical round clumping of tonofilaments, and in the other a whisk-type clumping of tonofilaments. Cultured keratinocytes from the former produced round clumps of keratin filaments, but those from the latter did not. Review of previous reports of Dowling-Meara EBS revealed that cases could also be divided into two groups in terms of the type of tonofilament clumping at an ultrastructural level. The possibility of subtyping of Dowling-Meara EBS, and possible mechanisms of the blistering in this disease are discussed. PMID- 8427827 TI - Three cases of primary acquired melanosis of the conjunctiva as a manifestation of the atypical mole syndrome. AB - We report three patients with the atypical mole syndrome (AMS) [also known as dysplastic naevus or FAMMM syndrome] who presented with primary acquired melanosis (PAM). PAM is a melanocytic lesion of the conjunctiva which may progress to conjunctival melanoma. The association of this rare condition with the AMS phenotype in three individuals suggests that PAM may be a conjunctival manifestation of the AMS. PMID- 8427828 TI - A case of xeroderma pigmentosum complementation group F with neurological abnormalities. AB - We report a 48-year-old Japanese man suffering from xeroderma pigmentosum associated with mental retardation, cerebral atrophy and cerebellar ataxia. Cultured fibroblasts from an unexposed area of skin had reduced DNA repair capacity after UV irradiation, with higher sensitivity to UV than normal cells in colony-forming ability and host cell reactivation using herpes simplex virus. Genetic complementation tests by cell fusion with polyethylene glycol revealed that the patient belonged to group F. He died of bile duct cancer at the age of 50. This is the first report of an XP-F patient with neurological abnormalities. PMID- 8427829 TI - Allopurinol-induced toxic pustuloderma. AB - We report the case of a 67-year-old man who developed a pustular eruption, fever, neutrophilia and eosinophilia, following a short course of allopurinol. Toxic pustuloderma is an uncommon form of generalized pustular eruption with several characteristic clinical and pathological features. A number of drugs have been incriminated, but to our knowledge this is the first reported case of toxic pustuloderma occurring after the administration of allopurinol. PMID- 8427830 TI - Solitary fibrosing paraspinal plaque: solitary morphoea profunda? AB - Solitary morphoea profunda is a recently described morphological variant of localized scleroderma. Two of the five reported patients had a solitary fibrotic plaque in the paraspinal region. We report a 16-year-old boy with a solitary fibrotic paraspinal plaque which, on histological examination, showed dermal and subcutaneous sclerosis, with a polymorphous infiltrate including plasma cells and eosinophils. Laboratory tests were either negative or normal, except for mild peripheral blood eosinophilia. Antinuclear antibodies were not detected, and the patient had no evidence of Borrelia infection. Magnetic resonance imaging (MRI) showed that the process involved the subcutaneous tissue, but did not extend to the underlying bone. As half of the patients with this entity (three of six) now described have had a solitary fibrotic plaque in a paraspinal location, we suggest that it may be premature to classify all these cases as being a variant of morphoea. We propose the use of the descriptive term solitary fibrosing paraspinal plaque, until the aetiology or aetiologies of this condition are better understood. PMID- 8427831 TI - Non-invasive first trimester antenatal diagnosis. PMID- 8427832 TI - The British Journal of Obstetrics and Gynaecology. A brief history. PMID- 8427833 TI - The role of research in the training of an obstetrician and gynaecologist. PMID- 8427834 TI - Thrombosis prophylaxis in obstetrics and gynaecology. PMID- 8427835 TI - The effects of exercise induced tachycardia on the maternal electrocardiogram. AB - OBJECTIVE: To determine whether pregnancy has an effect on the maternal electrocardiographic response to stress. DESIGN: Eight maternal ECG parameters were screened before, during and after exercise induced tachycardia at 29 and 37 weeks gestation and at 6 weeks post partum for each participant. SETTING: General Hospital in Safed, Israel. SUBJECTS: Fifteen healthy, pregnant women aged 22 to 34 years with singleton pregnancies volunteered from the peripheral antenatal clinics. INTERVENTION: Exercise stress tests were performed on a Quinton 3000 treadmill stress test system according to a Modified Bruce Protocol, producing a heart rate of 85% of age predicted maximum. MAIN OUTCOME MEASURES: The ST segment, T-wave, QRS complex, axis, rhythm, disorders of impulse conduction, the P-Q and Q-T intervals were screened before, during and after the exercise test. RESULTS: The study results suggest an increase in the ST-segment changes induced by stress at 37 weeks of pregnancy compared to 29 weeks (ST-depression in 6 women out of 15 versus 2, P = 0.125, McNemar's Test for Correlated Proportions-Exact Test, two tailed; 95% CI, 0.042-0.492, 0.01 < P < 0.05) and to the post partum test (ST-depression in 6 women out of 15 versus 1, P = 0.125; 0.029-0.637, 0.01 < P < 0.05). No significant alteration of the other ECG parameters. No significant correlation between blood haemoglobin level, serum levels of sodium, potassium, calcium, glucose and 17 beta-oestradiol with the appearance of the ECG change. CONCLUSIONS: The increase in the exercise induced ST-segment abnormalities at 37 weeks of pregnancy may be another physiological effect of late pregnancy which has not been demonstrated previously. PMID- 8427836 TI - Angiotensin sensitivity predicts aspirin benefit in placental insufficiency. AB - OBJECTIVE: To determine whether the beneficial effects of aspirin in the treatment of Doppler umbilical placental insufficiency correlate with the maternal pressor response to angiotensin infusion. DESIGN: An open trial. SETTING: A tertiary referral obstetric service. PATIENTS: Women identified at between 25 and 36 weeks of pregnancy with an elevated umbilical artery Doppler systolic/diastolic (S/D) ratio and a positive pressor response to angiotensin infusion. INTERVENTION: Low dose aspirin (100mg/day) treatment of mothers. MAIN OUTCOME MEASURE: Fetal and placental size at delivery in relation to subsequent maternal angiotensin responsiveness. RESULTS: Women who converted from a positive angiotensin pressor response to angiotensin refractoriness after aspirin administration had larger infants and placentas compared with those whose response remained positive. CONCLUSION: Angiotensin sensitivity predicts women with umbilical Doppler detected placental insufficiency responding to aspirin therapy. Loss of the normal refractory response in pregnancy may be a consequence of vascular pathology in the placenta. PMID- 8427837 TI - The present status of the Royal College of Obstetricians and Gynaecologists (excerpt from the address to new fellows and members on 5 June 1992). PMID- 8427838 TI - The loss of a baby at birth: the role of the bereavement officer. AB - OBJECTIVE: To illustrate and emphasise the role of the Bereavement Officer in the management of perinatal death, as recommended and envisaged by a report of the Royal College of Obstetricians and Gynaecologists in 1985. DESIGN: A description of the training and responsibilities of the officer and an account of her involvement with all perinatal deaths within a given period. SETTING: Perinatal deaths in the 12 months, 1 October 1989 to 30 September 1990 from a regional neonatal intensive care unit within a maternity hospital. SUBJECTS: A total of 59 registerable deaths, three late terminations (severe fetal abnormality) and 12 late mid-trimester miscarriages were dealt with by the Bereavement Officer in this 12 month period. RESULTS: The introduction of this service to our unit has led to an improvement in our management of both perinatal and fetal deaths. CONCLUSION: The service has justified fully the recommendations made by the Royal College of Obstetricians and Gynaecologists. Reducing parents' distress will facilitate their grieving and any subsequent bereavement counselling. PMID- 8427839 TI - Screening for fetal distress in labour using the umbilical artery blood velocity waveform. AB - OBJECTIVE: To investigate the potential value of umbilical artery blood velocity wave-form measurement as a screening test for intrapartum fetal distress on admission to the labour ward. DESIGN: Prospective study drawn from the local population of pregnant women. SETTING: The labour ward of the Princess Anne Hospital, Southampton, UK. SUBJECTS: 334 women with singleton pregnancies of at least 37 weeks gestation. MAIN OUTCOME MEASURE: Emergency caesarean section for fetal distress. RESULTS: There was a twelve-fold increase in the rate of emergency caesarean section for fetal distress (95% Confidence Interval (CI), 4.9 29) among women with a systolic/diastolic (S/D) ratio > or = 3.0, when compared to those with an S/D ratio of < 3.0 (P < 0.00001). Measurement of the umbilical artery blood velocity waveform compared favourably with admission cardiotocography. CONCLUSIONS: Umbilical artery blood velocity waveform analysis may be used to screen for fetal distress in labour and appears to be particularly sensitive to problems of placental origin. However, it is not likely to confer benefit in labour wards whose fetal heart rate monitoring policy is determined by pregnancy risk factors and admission cardiotocography. PMID- 8427840 TI - Abnormal fetal immunological development in Down's syndrome. AB - OBJECTIVE: To investigate the intrauterine development of the immune system in Down's syndrome. DESIGN: Cross sectional study. SETTING: Harris Birthright Research Centre for Fetal Medicine, London, UK. SUBJECTS: 16 fetuses with Down's syndrome and 104 fetuses with a normal karyotype at 17-24 weeks gestation. MAIN OUTCOME MEASURE: Flow cytometry was used to enumerate T (CD3+), B (CD19+) and NK (CD3- & CD16+/CD56+) lymphocyte subpopulations in fetal blood obtained by cordocentesis. RESULTS: The median numbers of T, B and NK lymphocytes in fetuses with Down's syndrome (1.52 x 10(9)/l, 0.08 x 10(9)/l, and 0.10 x 10(9)/l, respectively) were significantly lower than in the chromosomally normal fetuses (T lymphocytes: 1.98 x 10(9)/l, z = 3.04, P < 0.01; B lymphocytes: 0.50 x 10(9)/l, z = 5.84, P < 0.0001; and NK lymphocytes: 0.19 x 10(9)/l, z = 3.14, P < 0.01). CONCLUSION: These data demonstrate that in Down's syndrome, there is abnormal intra-uterine development of the immune system. PMID- 8427841 TI - Multifetal pregnancy reduction: a consecutive series of 61 cases. AB - OBJECTIVE: The effect of selective fetocide on the course of 61 multiple pregnancies. DESIGN: An observational study. SETTING: A tertiary centre. SUBJECTS: 61 women whose pregnancies included 37 triplets, 18 quadruplets, 5 quintuplets and 1 hepatuplet; 97% followed IVF or the induction of ovulation. The aim of the procedure in most cases was to obtain twins. INTERVENTIONS: Selective reduction was performed before 13 weeks gestation under general anaesthesia, using either a transcervical (n = 26) or transabdominal approach (n = 35). Fifty four twins, 4 singletons and 3 triplets were obtained after the procedure. MAIN OUTCOME MEASURE: Preterm labour rate. RESULTS: The rate of unplanned fetal loss was 13% and was related to the number of suppressed embryos (P < 0.05). The preterm labour rate was 56.6%, the mean gestation at delivery was 35.6 weeks. Seven deliveries were before 32 weeks and led to all neonatal deaths. A comparison with published data suggested that fetal reduction reduced the rate of preterm labour in high multiple pregnancies; in 24 twin pregnancies obtained after reduction of triplets there was probably a gain of 2 weeks gestation. Severe growth retardation occurred in 13%. The perinatal mortality rate was 10.8%. CONCLUSIONS: Selective termination reduces but does not prevent early preterm labour. The procedure is of value in pregnancies with more than 3 fetuses and should be considered carefully for triplet pregnancies. PMID- 8427842 TI - Prostaglandin prophylaxis and bladder function after vaginal hysterectomy: a prospective randomised study. AB - OBJECTIVE: To assess the efficacy of prostaglandins in enhancing bladder function after vaginal hysterectomy. DESIGN: Prospective randomised study of women who underwent vaginal hysterectomy between November 1989 and August 1990. SETTING: Sackler School of Medicine Tel Aviv University Medical Center, Department of Obstetrics and Gynaecology 'B'. SUBJECTS: 24 women who underwent vaginal hysterectomy and anterior and posterior colporrhaphy for prolapse. INTERVENTION: Administration of prostaglandin F2 alpha 5 mg intravesically or E2 3 mg intravaginally versus 100 ml saline intravesically (control) daily, starting on the first postoperative day, until adequate spontaneous voiding was established. RESULTS: Women receiving PGE2 resumed spontaneous bladder functions earlier, required significantly fewer days catheterisation and had significantly less febrile morbidity compared to women in the control group or PGE2 group. CONCLUSIONS: Prostaglandins PGE2 intravaginally are beneficial in enhancing functions after vaginal hysterectomy [corrected]. PMID- 8427843 TI - Psychosexual dysfunction in women with gynaecological cancer following radical pelvic surgery. AB - OBJECTIVE: To assess the prevalence and severity of psychosexual dysfunction in women treated for cancer of the cervix and vulva by radical vulvectomy, Wertheim's hysterectomy and pelvic exenteration; and to identify the risk factors for sexual morbidity and ways in which it might be reduced. DESIGN: Retrospective study of patients by questionnaire and semistructured interview, 6 months to 5 years following surgery. SETTING: Gynaecology-Oncology Unit of a general hospital. PATIENTS: 105 English speaking women with gynaecological cancer. RESULTS: 90% of the women in relationships had been sexually active prior to surgery. Of this group, 24% had no sexual difficulties post-operatively; 66% of the latter still had problems more than 6 months later, and 15% of the latter never resumed intercourse (excluding those with a colpectomy). 82% of those aged less than 50 years who had had radiotherapy suffered sexual dysfunction. Lack of desire was the commonest problem, and half the women felt that their sexual relationship had deteriorated, yet only 16% felt that their marriage had worsened. Younger women were more likely to attribute personal and marital distress to their sexual problems. More information on sexual matters would have been liked by 28% of the women. CONCLUSIONS: Sexual dysfunction is common following radical pelvic surgery and tends to remain a chronic problem. As well as organic causes there is a strong psychogenic element brought about by loss of fertility, disfigurement, depression and anxiety about one's desirability as a sexual partner. The presence of a stable relationship before the diagnosis of cancer helps women cope better, and young single women are a very vulnerable group. Patients want more information on sexual matters and the provision of sexual counselling may improve outcome in the future. PMID- 8427844 TI - Contact factor mediated fibrinolysis is increased by the combined oral contraceptive pill. AB - OBJECTIVE: To study the fibrinolytic pathways and their relationship with the contact system in women using combined oral contraceptives (COCs). DESIGN: Serial plasma samples were collected from 18 women before treatment with COCs containing 30 micrograms oestrogen during treatment cycles 3 and 6, and 2 weeks after stopping treatment. Fibrinolysis was measured before and after dextran sulphate mediated contact activation using fibrin plates. RESULTS: Fibrinolysis increased significantly during cycles 3 and 6 (from 77% to 100% and 113%, respectively, P < 0.01) and showed a further increase after dextran sulphate activation (from 134% to 158% and 167%, respectively, P < 0.01). Tissue-plasminogen activator, urokinase-plasminogen activator and plasminogen activator inhibitor did not change significantly. There were significant elevations of Factor XII (from 0.92 u/ml to 1.43 u/ml, P < 0.01) and prekallikrein (0.94 u/ml to 1.10 u/ml, P < 0.05) in cycle 3, which both remained high at cycle 6 (P < 0.01) and decreased after stopping the COC. Alpha-2-macroglobulin and C1-esterase inhibitor showed no significant change, but alpha-1-antitrypsin increased from 0.85 u/ml to 1.11 u/ml by cycle 3 (P < 0.01), and returned to near normal levels after stopping the COC. CONCLUSIONS: The increase in fibrinolysis may be due to increased levels of Factor XII and prekallikrein without a corresponding increase in their natural inhibitors (C1-esterase inhibitor and alpha-2-macroglobulin). A parallel increase in the intrinsic pathway of coagulation may be limited by elevated alpha-1 antitrypsin at the level of activated Factor XI. The increase in fibrinolysis caused by oral contraceptives may balance any potential thrombotic risk due to increased fibrinogen or vitamin K dependent coagulation factors. PMID- 8427845 TI - Noninvasive assessment of the maternal cerebral circulation by transcranial Doppler ultrasound during angiotensin II infusion. AB - OBJECTIVE: To examine the relationship between experimentally induced increments in blood pressure and maternal middle cerebral artery flow velocity patterns measured by transcranial Doppler ultrasound (TCD). DESIGN: Prospective experimental study. SETTING: John Radcliffe Maternity Hospital, Oxford. INTERVENTION: Middle cerebral artery flow velocity waveforms were measured using 2 MHz pulsed Doppler ultrasound via the temporal cranial approach at 28 weeks gestation before, during and 10 min following an angiotensin II infusion. SUBJECTS: 101 normotensive primiparous women at 28 weeks gestation. RESULTS: A significant rise in blood pressure and fall in heart rate were demonstrated between pre-infusion and maximum angiotensin II infusion (maximum blood pressure) recordings. Simultaneous changes were observed in all flow velocity indices, shown as a decrease in systolic velocity and pulsatility index, and an increase in diastolic and mean velocity (P < 0.0001). These values all returned to baseline levels 10 min post infusion (P < 0.0001). Statistical analysis suggested that the change in flow velocity is related to the rise in blood pressure rather than the direct effect of angiotensin II on the cerebral circulation. CONCLUSION: Transcranial Doppler can detect changes in the cerebral circulation associated with alterations in blood pressure during pregnancy. The technique needs to be evaluated further in hypertensive disease. PMID- 8427846 TI - Rapid DNA quantification in the prenatal diagnosis of fetal triploidy. PMID- 8427847 TI - A prospective randomised controlled trial of perineal repair after childbirth, comparing interrupted chromic catgut to subcuticular prolene for skin closure. PMID- 8427848 TI - Successful treatment of mammary hyperplasia in pregnancy with bromocriptine. PMID- 8427849 TI - Renal failure in two preterm infants: toxic effect of prenatal maternal indomethacin treatment? PMID- 8427850 TI - Randomised trial of amniotomy in labour versus the intention to leave membranes intact until second stage. PMID- 8427851 TI - Effect of hepatic lipase on LDL in normal men and those with coronary artery disease. AB - Hepatic triglyceride lipase (HL) is thought to play a role in the formation of low density lipoproteins (LDLs) from small very low density lipoproteins (VLDLs) and intermediate density lipoproteins (IDLs). To analyze the possible physiological role of HL in determining LDL buoyancy, size, and chemical composition, HL activity and LDL were studied in 21 patients with coronary artery disease (CAD) and 23 normolipidemic subjects. In both groups, LDL buoyancy and size were inversely associated with HL activity levels. The effect of HL on LDL size was comparable in CAD patients and in normolipidemic subjects. HL appeared to influence LDL lipid composition primarily by affecting the surface lipid components. The free cholesterol content of LDL particles was highly correlated with HL activity in both CAD and normolipidemic individuals. The free cholesterol to phospholipid ratio in LDL particles correlated with HL in both CAD and normolipidemic subjects. When the individuals were separated according to their LDL subclass patterns, pattern B subjects had significantly higher HL than pattern A subjects in both CAD and normolipidemic groups. The analysis of the cholesterol distribution profiles across the lipoprotein density gradient confirmed that LDL buoyancy is affected by HL. These data support the hypothesis that HL modulates the physical and compositional properties of LDL and contributes to the expression of the LDL subclass phenotype, suggesting a physiological role for HL in LDL metabolism. PMID- 8427852 TI - Hydrogenation impairs the hypolipidemic effect of corn oil in humans. Hydrogenation, trans fatty acids, and plasma lipids. AB - The effects of plasma lipoproteins and apolipoproteins of replacing corn oil with corn-oil margarine in stick form as two thirds of the fat in the National Cholesterol Education Program (NCEP) Step 2 diet were assessed in 14 middle-aged and elderly women and men (age range, 44-78 years) with moderate hypercholesterolemia (low density lipoprotein cholesterol [LDL-C] range, 133-219 mg/dl [3.45-5.67 mmol/l] at screening). During each 32-day study phase, subjects received all their food and drink from a metabolic kitchen. Subjects were first studied while being fed a diet approximating the composition of the current US diet (baseline), which contained 35% of calories as fat (13% saturated fatty acids [SFAs], 12% monounsaturated fatty acids [MUFAs; 0.8% 18:1n-9 trans], and 8% polyunsaturated fatty acids [PUFAs]) and 128 mg cholesterol/1,000 kcal. This baseline phase was followed by a corn oil-enriched diet containing 30% fat (6% SFA, 11% MUFA [0.4% 18:1n-9 trans], and 10% PUFA) and 83 mg cholesterol/1,000 kcal, and then a corn-oil margarine-enriched diet containing 30% fat (8% SFA, 12% MUFA [4.2% 18:1n-9 trans], and 8% PUFA) and 77 mg cholesterol/1,000 kcal. All diets were isocaloric. Mean fasting LDL-C and apolipoprotein (apo) B levels were 153 mg/dl (3.96 mmol/l) and 101 mg/dl on the baseline diet, 17% and 20% lower (both p < 0.001) on the corn oil-enriched diet, and 10% and 10% lower (both p < 0.01) on the margarine-enriched diet.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427853 TI - Impact of weight loss on plasminogen activator inhibitor (PAI-1), factor VII, and other hemostatic factors in moderately overweight adults. AB - Based on previous cross-sectional findings, we hypothesized that weight loss could improve several hemostatic factors associated with cardiovascular disease. In a randomized controlled trial, moderately overweight men and women were assigned to one of four weight loss treatment groups or to a control group. Measurements of plasminogen activator inhibitor-1 (PAI-1) antigen, tissue-type plasminogen activator (t-PA) antigen, D-dimer antigen, factor VII activity, fibrinogen, and protein C antigens were made at baseline and after 6 months in 90 men and 88 women. Net treatment weight loss was 9.4 kg in men and 7.4 kg in women. There was no net change (p > 0.05) in D-dimer, fibrinogen, or protein C with weight loss. Significant (p < 0.05) decreases were observed in the combined treatment groups compared with the control group for mean PAI-1 (31% decline), t PA antigen (24% decline), and factor VII (11% decline). Decreases in these hemostatic variables were correlated with the amount of weight lost and the degree that plasma triglycerides declined; these correlations were stronger in men than women. These findings suggest that weight loss can improve abnormalities in hemostatic factors associated with obesity. PMID- 8427854 TI - Effects of simvastatin on apoB metabolism and LDL subfraction distribution. AB - Seven moderately hypercholesterolemic subjects were studied before and after 10 weeks of simvastatin therapy (20 mg/day). Therapy reduced low density lipoprotein (LDL) cholesterol by 39% (p < 0.001), whereas high density lipoprotein and very low density lipoprotein (VLDL) cholesterol were unchanged. Apolipoprotein (apo) B containing lipoproteins were divided into VLDL1 (Sf 60-400), VLDL2 (Sf 20-60), intermediate density lipoprotein (IDL) (Sf 12-20), and LDL (Sf 0-12), and metabolic changes were sought in dual-tracer VLDL1 and VLDL2 turnover studies. VLDL1 apoB pool size was unaltered by therapy, as were its rates of synthesis, catabolism, and delipidation to VLDL2. Similarly, the VLDL2 apoB pool size was unchanged, but its metabolic fate was altered. The IDL pool size fell significantly (27%, p < 0.01) due entirely to an increased fractional catabolism of the lipoprotein. In our subjects, the circulating mass of LDL apoB decreased (49%, p < 0.01) primarily due to a reduction in its synthesis. Before therapy, 30% of the apoB entering the delipidation cascade in these hyperlipidemic subjects was converted to LDL. On therapy the input remained the same, but direct catabolism from VLDL2 and IDL was increased (p < 0.05), and as a result only 16% eventually appeared in LDL. These kinetic changes were associated with a fall in particle cholesteryl ester content throughout the delipidation cascade. We also observed a link between LDL kinetics and its subfraction distribution. Simvastatin influences the metabolism of LDL, IDL, and VLDL2 but not VLDL1. PMID- 8427855 TI - High macrophage lipoprotein lipase expression and secretion are associated in inbred murine strains with susceptibility to atherosclerosis. AB - To test the possibility that variations in macrophage lipoprotein lipase (LPL) secretion may constitute one of the hereditary components of atherosclerosis, we evaluated LPL gene expression and secretion in macrophages harvested from inbred mouse strains differing in their susceptibility to the diet-induced development of atherosclerosis. Inflammatory peritoneal macrophages harvested from atherosclerosis-susceptible C57BL/6J mice showed twofold to threefold higher basal LPL mass, activity, and mRNA levels than those isolated from atherosclerosis-resistant C3H/HeN mice. We determined LPL secretion and gene expression in the susceptible C57BL/6J (B), resistant A/J (A), and A x B/B x A recombinant inbred strains of mice typed as atherosclerosis resistant (A-like) or atherosclerosis susceptible (B-like). Macrophage LPL secretion and mRNA expression were twofold higher in the susceptible C57BL/6J (B) mice than in the resistant A/J (A) mice. Significantly higher LPL secretion, activity, and gene expression were found in recombinant inbred mouse strains that typed B-like than in those typed A-like. These results indicate that susceptibility to atherosclerosis is associated in inbred mouse strains with high LPL secretion and mRNA levels, whereas lower LPL secretion and mRNA expression are observed in atherosclerosis-resistant mice. These observations suggest a contributive role for LPL in the development of atherosclerosis. PMID- 8427856 TI - Cardiovascular risk factors in non-insulin-dependent diabetic subjects with microalbuminuria. AB - In subjects with insulin-dependent diabetes mellitus, microalbuminuria has been associated with increased triglyceride and lipoprotein (a) (Lp[a]) concentrations and increased blood pressure. However, few studies have examined whether this association is present in subjects with non-insulin-dependent diabetes mellitus (NIDDM). We measured lipids, lipoproteins, Lp(a), blood pressure, and albumin excretion in 234 subjects with NIDDM from the San Antonio Heart Study, a population-based study of diabetes and cardiovascular disease. Seventy-two subjects had microalbuminuria (> or = 30 mg/dl). These subjects had increased systolic and diastolic blood pressures and higher fasting glucose concentrations relative to subjects without microalbuminuria. However, there were no significant differences between subjects with and without microalbuminuria with respect to lipids, lipoproteins, Lp(a), self-reported myocardial infarction, obesity, or body fat distribution. Subjects with diabetic retinopathy had increased microalbuminuria. In multivariate analysis both glycemia and blood pressure continued to be significantly related to the presence of microalbuminuria. We conclude that NIDDM subjects with microalbuminuria have elevated blood pressure and more severe glycemia but do not have a significantly more atherogenic pattern of lipids, lipoproteins, or Lp(a) than subjects without microalbuminuria. PMID- 8427858 TI - Interactions between the monocyte/macrophage and the vascular smooth muscle cell. Stimulation of mitogenesis by a soluble factor and of prostanoid synthesis by cell-cell contact. AB - The effect of soluble factors from the monocyte/macrophage (M phi) on cell proliferation and the functional effects of cell-cell contact on the arachidonic acid (AA) cascade were studied with vascular smooth muscle cells (SMCs). Peripheral blood M phi s were isolated by adherence or in a Percoll gradient, and alveolar M phi s were obtained by lavage. Conditioned medium (CM) was prepared by preincubating M phi s with medium alone or by separating SMC and M phi cocultures by a membrane insert. Cell proliferation (image analysis) and 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha, radioimmunoassay) were measured in SMCs. Labeled prostanoids and other eicosanoid metabolites were isolated by high-performance liquid chromatography from SMCs prelabeled with 14C-AA. M phi s did not synthesize 6-keto-PGF1 alpha. The CM enhanced proliferation but did not stimulate 6-keto-PGF1 alpha synthesis in SMCs. However, cell-cell contact in cocultures of SMCs with the same concentration of M phi s used to generate CM resulted in increased 6-keto-PGF1 alpha synthesis by SMCs. Since the stimulatory effect of cell contact was not blocked by butylated hydroxytoluene, it could not be attributed to an oxidative burst from M phi s. Functional studies showed that the stimulatory effect of cell contact was enhanced by exogenous free AA and by endogenous AA release through A23187. Release of total radioactivity from prelabeled SMCs was enhanced by cell contact, and this effect was blocked by indomethacin (IM). Cell contact did not increase the release of free AA from prelabeled SMCs, even in the presence of IM. Finally, cell contact only stimulated the formation of prostanoids (IM-sensitive eicosanoid metabolites) from prelabeled SMCs. Lipoxygenase and other products of AA were not formed through cell-cell contact. These data showed that M phi s express a soluble factor that enhances SMC proliferation without affecting prostanoid synthesis. Subsequent cell contact between SMCs and M phi s stimulates prostanoid synthesis, which may possibly serve as a local and focal homeostatic mechanism for the regulation of uncontrolled SMC proliferation in atherogenesis. PMID- 8427857 TI - Smooth muscle cell immediate-early gene and growth factor activation follows vascular injury. A putative in vivo mechanism for autocrine growth. AB - To understand the molecular events governing smooth muscle cell (SMC) proliferation in vivo, immediate-early gene (IEG) expression was assessed and related to growth factor ligand and receptor mRNA and SMC DNA synthesis after aortic injury. Balloon catheter injury evoked increases in SMC c-myc and thrombospondin (tsp) within 2 hours. The induction of these IEGs was followed by elevated transcripts to platelet-derived growth factor-A (PDGF-A), transforming growth factor-beta 1 (TGF-beta 1) and a basic fibroblast growth factor (bFGF) receptor. Whereas PDGF type-beta receptor mRNA was demonstrated in SMCs from control and balloon-injured aortas, no detectable signal was observed for the PDGF type-alpha receptor. To explore the potential linkage between IEG products and growth factor mRNA expression, cycloheximide was employed to block early protein synthesis after balloon injury. Induction of PDGF-A and TGF-beta 1 was attenuated by cycloheximide, but bFGF induction was unaffected. Moreover, cycloheximide superinduced IEGs and revealed PDGF-B transcripts, which were otherwise undetected. Seven days after aortic injury, a spontaneous increase in c myc and tsp mRNA was noted. This IEG reactivation was followed 12 hours later by a twofold increase in SMC DNA synthesis. These findings corroborate an autocrine mode of SMC proliferation in vivo and suggest the IEG products may control such growth by stimulating growth factor genes. PMID- 8427859 TI - Glutamine/histidine polymorphism in apo A-IV affects plasma concentrations of lipoprotein(a) and fibrin split products in coronary heart disease patients. AB - A glutamine/histidine polymorphism at residue 360 in apolipoprotein (apo) A-IV that generates two electrophoretically detectable isoforms, apo A-IV-1 and apo A IV-2, affects the plasma concentration of lipoprotein(a) (Lp[a]) in a healthy population. To verify this unexpected association we analyzed the effect of the apo A-IV polymorphism on Lp(a) serum concentrations in 275 male coronary heart disease patients. Allele frequencies of apo A-IV-1 and apo A-IV-2 were 0.917 and 0.083, respectively. In addition, apo A-IV-1/2 heterozygotes showed a 30% lower geometric mean concentration of Lp(a) than apo A-IV-1/1 homozygotes in this study. The relative frequency of Lp(a) concentrations > 20 mg/dl was significantly increased by a factor of 2.25 in apo A-IV-1/1 homozygotes. Other lipid parameters were not significantly affected by this apo A-IV polymorphism. Because of the relations between Lp(a) and the fibrinolytic system, we also analyzed the effect of the apo A-IV polymorphism on hemostatic variables. Apo A IV-1/2 heterozygosity was associated with a 70% higher geometric mean plasma concentration of D-dimer, i.e., proteolytic fragments of cross-linked fibrin. Plasma concentrations of prothrombin fragments F1 + F2, fibrinogen, plasminogen, and plasminogen activator inhibitor-1 were unaffected. In conclusion, our results indicate a hitherto unappreciated role of the apo A-IV gene or a closely linked locus for the regulation of Lp(a) metabolism and hemostasis and also possibly for atherosclerosis and thrombosis. PMID- 8427860 TI - Plaque changes and arterial enlargement in atherosclerotic monkeys after manipulation of diet and social environment. AB - To study the effects of dietary and social manipulations on lesion progression in male monkeys with established atherosclerosis, 83 animals fed a diet containing 1 mg cholesterol per kcal for 14 months were either necropsied (baseline group, n = 21) or assigned to one of three experimental conditions: 1) a diet containing a high amount of fat and cholesterol and a stressful social situation (HiFC-stress, n = 18); 2) a diet lower in fat and cholesterol and a stressful social situation (LoFC-stress, n = 21); or 3) the low-fat, low-cholesterol diet and a nonstressful social situation (LoFC-no stress, n = 23). After 28 months, all animals were necropsied. Coronary atherogenesis was arrested among monkeys in the LoFC-stress and LoFC-no stress conditions compared with that of animals in the baseline condition (plaque areas of 0.35 mm2, 0.30 mm2, and 0.38 mm2, respectively). Lesions in animals fed the LoFC diet (both stress and no-stress groups) were significantly smaller than those in monkeys in the HiFC-stress condition (0.96 mm2). Furthermore, aortic cholesterol content was significantly decreased and luminal areas were relatively larger among monkeys in both LoFC conditions compared with animals in the baseline and HiFC-stress conditions (p < 0.05 for all). The results demonstrate that a low-fat, low-cholesterol diet can halt plaque development, reduce arterial cholesterol content, and permit compensatory arterial enlargement, processes that were unaffected by social stress in this investigation. PMID- 8427861 TI - Cholesteryl ester loading of mouse peritoneal macrophages is associated with changes in the expression or modification of specific cellular proteins, including increase in an alpha-enolase isoform. AB - This report explores the hypothesis that massive cholesteryl ester (CE) accumulation in macrophages, such as that occurring in atheroma foam cells, results in changes in the expression or modification of specific cellular proteins. Two-dimensional (2-D) gel electrophoretic patterns of metabolically labeled cellular proteins from mouse peritoneal macrophages that were loaded with CE (through incubation with acetylated low density lipoprotein [acetyl-LDL] for 4 days) were compared with those of control macrophages. Densitometric analysis of 2-D gel autoradiograms from the cell lysates revealed statistically significant changes in seven cellular proteins (five decreases and two increases). The changes in protein expression (foam cell versus control) ranged from a 458 +/- 164% (p < 0.001) increase to a 35 +/- 34% (p < 0.001) decrease (n = 11). Incubation of macrophages with beta-very low density lipoprotein, which also increased the CE content of macrophages (albeit to a lesser extent than acetyl LDL), resulted in changes in five of the seven proteins. In contrast, incubation of cells with LDL, fucoidan, or latex beads, none of which caused CE accumulation, did not lead to significant changes in four of these five proteins. One of these four proteins, which increased fourfold to fivefold in foam cells (M(r) = 49,000; isoelectric point of 6.8), was purified by preparative 2-D gel electrophoresis. Internal amino acid sequence of cyanogen bromide fragments of this protein as well as Western blot analysis identified this protein as an isoform of alpha-enolase. The increased expression of this alpha-enolase isoform, which was seen as early as day 2 of acetyl-LDL incubation of the macrophages, was diminished by including an inhibitor of cholesterol esterification during the acetyl-LDL incubation period. In conclusion, macrophage foam cell formation is associated with distinct changes in protein expression, including a marked increase in an isoform of alpha-enolase, suggesting a specific biological adaptation to CE loading. PMID- 8427862 TI - Metabolism of LDL in mast cells recovering from degranulation. Description of a novel intracellular pathway leading to proteolytic modification of the lipoprotein. AB - Rat serosal mast cells contain cytoplasmic secretory granules composed of a proteoglycan matrix in which histamine and neutral proteases are embedded. On stimulation, these granules are exocytosed, but some of them remain in the degranulation channels where on exposure to the extracellular fluid, they lose their histamine and a fraction of their proteoglycans. In vitro, such granule remnants efficiently bind low density lipoprotein (LDL) present in the incubation medium. After a lag period of about 10 minutes, the granule remnants, still within the channels and coated with LDL particles, are internalized by the parent mast cells. During subsequent recovery from degranulation, the apolipoprotein B of the intracellularly located remnant-bound LDL becomes efficiently (up to 70%) degraded by the proteolytic enzymes of the granule remnants. Since the granule remnants lack cholesteryl esterase activity, no LDL cholesterol is made available for cellular nutrition. Instead, selective proteolytic degradation of the bound LDL leads to formation of LDL particles enlarged by fusion on the granule remnant surface. In response to restimulation of the mast cells, about 50% of the fused LDL particles are exocytosed with the granule remnants. Of these, about one in five are expelled into the incubation medium. The granule remnants that again remain in the degranulation channels bind and internalize more LDL. This "round trip" of LDL in mast cells exposed to repeated stimulation constitutes a hitherto unknown intracellular pathway for modification of LDL. PMID- 8427863 TI - Gemfibrozil reduces postprandial lipemia in non-insulin-dependent diabetes mellitus. AB - The effect of gemfibrozil on postprandial lipoprotein metabolism was investigated in a 12-week, randomized, double-blind, placebo-controlled trial in 20 non insulin-dependent diabetic patients with moderate hypertriglyceridemia. The patients were given a meal containing 78 g of fat and 345,000 units of vitamin A to label chylomicrons and their remnants. Plasma obtained at various times during the fat-load test was separated into six fractions by gradient-density ultracentrifugation. Gemfibrozil reduced the postprandial triglyceride response, measured as the area under the time-dependent concentration curve, on average by 32% in whole plasma, by 38% in the Svedberg flotation unit (Sf) 1,100-3,200 chylomicron fraction, by 36% in Sf 400-1,100 chylomicrons, and by 38% in the Sf 60-400 lipoproteins. Retinyl palmitate, a measure of intestinally derived particles, was reduced in plasma by 34%, in Sf 1,100-3,200 by 46%, in Sf 400 1,100 by 44%, and in Sf 60-400 by 37%. All these reductions were significant in comparison with the placebo group. Particles with Sf < 60 were not significantly affected. In contrast to earlier observations in healthy subjects, no significant negative correlations existed between postprandial lipemia and high density lipoprotein cholesterol or the postheparin lipoprotein lipase activity. The reduction of the potentially atherogenic chylomicron remnants may decrease the risk of atherosclerosis in non-insulin-dependent diabetes mellitus, a hypothesis that awaits testing in prospective studies. PMID- 8427864 TI - Growth hormone treatment of growth hormone-deficient adults results in a marked increase in Lp(a) and HDL cholesterol concentrations. AB - The effects of growth hormone treatment of adults with adult-onset pituitary insufficiency on lipoproteins and apolipoproteins were investigated. Nine patients, one women and eight men (age range, 34-58 years), who had been treated for pituitary tumors were studied. They had complete pituitary insufficiency with a duration of at least 1 year. All patients received replacement therapy with thyroid hormones, glucocorticoids, and gonadal steroids. The study had a double blind, placebo-controlled, crossover design for active treatment with recombinant human growth hormone (0.25-0.5 units/kg per week s.c. given each evening) for 6 months. Fasting serum levels of cholesterol; triglycerides; high density lipoprotein and low density lipoprotein cholesterol; apolipoproteins A-I, B, and E; and lipoprotein (a) were measured before and after 6 and 26 weeks of treatment. Lipoprotein (a) concentrations increased markedly during treatment and were about twice as high compared with pretreatment levels. Serum cholesterol and low density lipoprotein cholesterol concentrations were decreased after 6 weeks of treatment, but levels had returned to pretreatment levels after 26 weeks. High density lipoprotein cholesterol concentrations increased during treatment and were significantly higher than pretreatment levels after 26 weeks of treatment. Serum triglyceride concentrations did not change significantly, but in two patients with marked hypertriglyceridemia, growth hormone treatment resulted in a marked decrease. Serum concentrations of apolipoproteins A-I, B, and E did not change significantly, but changes in apolipoprotein A-I and B concentrations were in parallel to those observed for high density lipoprotein cholesterol and low density lipoprotein cholesterol, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427865 TI - Lipoprotein profile characterization of the KKA(y) mouse, a rodent model of type II diabetes, before and after treatment with the insulin-sensitizing agent pioglitazone. AB - The purpose of this study was to characterize the lipoprotein profile in the KKA(y) mouse, a rodent model of type II diabetes, before and after treatment with the insulin-sensitizing drug pioglitazone. Analysis of the plasma from untreated KKA(y) mice showed that they were severely hyperglycemic, severely hypertriglyceridemic, and moderately hypercholesterolemic. Agarose column chromatographic analysis showed that essentially all of the triglyceride eluted with very low density lipoprotein, and the majority of the cholesterol eluted with high density lipoprotein. Thus, both the very low density lipoprotein and high density lipoprotein levels were markedly elevated in KKA(y) mice. Analysis of the lipoproteins by agarose electrophoresis-immunoblotting showed that apoprotein A-I and apoprotein B had aberrant electrophoretic behavior, typical of apoproteins that have been modified by nonenzymatic glycosylation. Treatment of KKA(y) mice with pioglitazone for 8 days caused a marked reduction in blood glucose and plasma triglyceride concentrations but had no significant effect on plasma cholesterol concentration or distribution. The aberrant electrophoretic behavior of the apoproteins was corrected to normal by drug treatment. These data show that the KKAy mouse has a severe dyslipoproteinemia that is probably secondary to its insulin resistance, but that its lipoprotein profile differs significantly from that of the insulin-resistant human in that the majority of the plasma cholesterol is carried in high density lipoprotein, and those high density lipoprotein levels are very high. PMID- 8427866 TI - Relation of vessel wall shear stress to atherosclerosis progression in human coronary arteries. AB - The purpose of this study was to determine the relation between vessel wall shear stress and the rate of atherosclerosis progression. Quantitative angiography was used to calculate the change in coronary arterial diameter over 3.0 years in patients enrolled in the Harvard Atherosclerosis Reversibility Project pilot study (n = 20 arterial segments). Vessel wall shear stress was calculated by means of a validated finite-difference model of the Navier-Stokes' equation that assumes a coronary flow rate of 8 ml/sec. The correlation between vessel wall shear stress and the change in arterial diameter at multiple points (mean, 70) along the length of the artery was then calculated for each of the 20 segments with a focal stenosis. In 15 of the 20 arterial segments there was a significant correlation (p < 0.05) between low shear stress and an increased rate of atherosclerosis progression. A Fisher's z transformation was then used to combine the correlation coefficients from all 20 segments. Low shear stress was significantly correlated (z = 0.37 +/- 0.00074, p < 0.0001) with an increased rate of atherosclerosis progression. This serial quantitative evaluation of human coronary arteries is consistent with previous data that have suggested that low shear stress promotes atherosclerosis progression. Variations in local vessel wall shear stress may explain the previously reported near-independent rate of atherosclerosis progression in multiple lesions within the same patient despite exposure to the same circulating lipoprotein values and systemic hemodynamics. PMID- 8427867 TI - Lipoproteins in familial dysbetalipoproteinemia. Variation of serum cholesterol level associated with VLDL concentration. AB - Patients with familial dysbetalipoproteinemia (FD) associated with the apo E2/2 phenotype exhibit a marked interindividual variability in serum cholesterol and triglyceride concentrations. It has been proposed that this variability is due to a combination of the apo E2/2 phenotype and additional genetic factors implicated in diseases like familial hypercholesterolemia, familial combined hyperlipoproteinemia, and familial hypertriglyceridemia. To further explore the nature of this variability, the lipoprotein profiles of 17 patients with FD associated with the apo E2/2 phenotype were analyzed by a density-gradient ultracentrifugation technique and by 2-16% polyacrylamide gel electrophoresis. It was found that all patients with FD were characterized by 1) markedly increased cholesterol concentrations of large very low density lipoprotein (VLDL) (VLDL1) (2.98 +/- 3.08 versus 0.08 +/- 0.03 mmol/L), small VLDL (VLDL2) (4.68 +/- 1.93 versus 0.27 +/- 0.13 mmol/L), and intermediate density lipoprotein (IDL) (2.25 +/ 0.72 versus 0.39 +/- 0.16 mmol/L); 2) decreased low density lipoprotein (LDL) cholesterol level (1.84 +/- 0.54 versus 3.36 +/- 0.53 mmol/L); and 3) altered composition (enrichment by cholesteryl ester) of VLDL1 and VLDL2 compared with normolipidemic control subjects. The cholesterol levels of IDL and LDL showed minor interindividual variabilities and were not correlated with serum cholesterol and triglyceride levels. The compositions of VLDL1 and VLDL2 were independent of the concentrations of lipids in serum. However, the cholesterol concentrations of VLDL1 and VLDL2 showed considerable interindividual variabilities and were positively correlated with the serum cholesterol concentration (r = 0.84 and r = 0.95, respectively, both p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8427868 TI - Immunological characterization of flavin-containing monooxygenases from the liver of rainbow trout (Oncorhynchus mykiss): sexual- and age-dependent differences and the effect of trimethylamine on enzyme regulation. AB - Polyclonal antibodies raised against flavin-containing monooxygenase (FMO) enzymes purified from pig liver and rabbit lung were used in conjunction with N,N dimethylaniline (DMA) N-oxidase to better characterize FMO from the liver of rainbow trout (Oncorhynchus mykiss). Two proteins reacted with polyclonal antibodies raised against pig liver FMO (PL-1 and PL-2) and anti-rabbit lung FMO (RL-1 and RL-2). Although there was no difference in DMA N-oxidase observed between sexually mature male and female trout liver microsomes, RL-2 and PL-2 were significantly less than RL-1 and PL-1, respectively, in sexually mature females. FMO activity and protein content increased as fish aged. DMA oxidase and FMO isozymes were unaltered after pretreatment with the endogenous substrate trimethylamine. Since antibodies to the purified mammalian enzymes react with proteins of similar MW in trout, some forms of FMO appear to be structurally conserved through evolution. PMID- 8427869 TI - Halofuginone: an inhibitor of collagen type I synthesis. AB - The effect of halofuginone--a plant alkaloid used as a coccidiostat in birds--on collagen metabolism was studied in various avian and mammalian cell cultures. In avian skin fibroblasts halofuginone attenuated the incorporation of [3H]proline into collagenase-digestible proteins (CDP) at concentrations as low as 10(-11) M, without affecting production of [3H]collagenase-nondigestible proteins (NCDP), cell proliferation or collagen degradation. Halofuginone depressed specifically the expression of alpha 1 gene of collagen type I but not that of collagen type II. This was demonstrated in skin fibroblasts and growth-plate chondrocytes using probes containing inserts sequences corresponding to the alpha 1(I) and alpha 1(II) mRNAs. A slight inhibition of the expression of alpha 2(I) was observed in avian skin fibroblasts but not in growth-plate chondrocytes. The inhibition of gene expression of both polypeptides of collagen type I in skin fibroblasts resulted in a decrease in synthesis, as demonstrated by immunoprecipitation with specific type I collagen antiserum. In primary cultures of mouse skin fibroblasts, avian epiphyseal growth plate chondrocytes and a rat embryo cell line--all of which produce and secrete collagen type I--halofuginone inhibited the incorporation of [3H]proline into CDP, the Rat-1 line being the most sensitive to the drug. These results suggest that halofuginone affects specifically type I collagen synthesis by repressing gene-expression. The need for extremely low concentrations of halofuginone to inhibit collagen type I synthesis, regardless of the tissue or animal species, contributes to the potential usefulness of the substance in studying collagen metabolism. PMID- 8427870 TI - Polyamine acetylation in rat liver following long-term ethanol ingestion. AB - The effect of ethanol on polyamine acetylation was studied in rat liver. Animals were fed on nutritionally complete liquid diets with 36% or 12% of total calories supplied as ethanol or isocaloric carbohydrates for 4 months. The diet with 36% calories as ethanol significantly increased the activity of cytosolic spermidine/spermine N1-acetyltransferase, the rate-limiting enzyme in polyamine interconversion. Such a stimulation did not appear in rats under the 12% ethanol regimen. The stimulation of spermidine/spermine N1-acetyltransferase was associated with increases in putrescine and spermidine concentrations, an appearance of N1-acetylspermidine, and a decrease in spermine. These results suggest that chronic ethanol intake stimulates the acetylation of polyamines and their interconversion in rat liver. PMID- 8427871 TI - Salvage and interconversion of purines in developing Artemia. AB - Incorporation of the radiolabelled purine bases adenine, guanine and hypoxanthine into acid soluble fraction, RNA and DNA nucleotides during the early larval development of Artemia sp. was studied. Adenine was the best precursor and guanine the poorest. The adenine phosphoribosyltransferase (APRT) activity was considerably higher than that of hypoxanthine-guanine phosphoribosyltransferase (HGPRT) and these activities did not significantly change throughout larval development. The pattern of purine interconversion was dependent on naupliar age. Conversion of [14C]adenine and [14C]hypoxanthine into guanine nucleotides increased with time of development. However, the conversion of [14C]guanine into [14C]adenine nucleotides was very low. PMID- 8427872 TI - A top-down control analysis in isolated rat liver mitochondria: can the 3-hydroxy 3-methylglutaryl-CoA pathway be rate-controlling for ketogenesis? AB - We incubated isolated liver mitochondria with palmitoyl-CoA, 2,4-dinitrophenol and malonate. Under these conditions all the flux of carbon from palmitoyl-CoA was directed towards acetoacetate synthesis. We measured the rate of acetyl-CoA formation from palmitoyl-CoA (by measuring the rate of oxygen consumption) and the rate of acetoacetate production from acetyl-CoA at three different acetyl CoA/CoA ratios. Using the top-down approach of metabolic control analysis we calculated the control over ketogenesis exerted by (a) the conversion of extramitochondrial palmitoyl-CoA to intramitochondrial acetyl-CoA and by (b) the conversion of acetyl-CoA to acetoacetate (the 'HMG-CoA pathway'). The overall flux control coefficients of the groups of enzymes involved in (a) and (b) over ketogenesis were 0.28 and 0.72, respectively. Our results show that it is possible for significant control to be exerted over ketogenesis by the enzymes of the HMG-CoA pathway. PMID- 8427873 TI - Antioxidant protection against organic and inorganic oxygen radicals by normal human plasma: the important primary role for iron-binding and iron-oxidising proteins. AB - Normal human plasma contains numerous high and low molecular mass redox active molecules that are able to react rapidly with organic and inorganic oxygen radicals. The ability of such plasma molecules to substantially inhibit, or delay, free radical mediated oxidation of added substrates has led to their classification as important biological antioxidants. Using phospholipids to detect organic oxygen radicals, and deoxyribose to detect inorganic oxygen radicals, we here show that the primary antioxidants of normal human plasma reside mainly in two plasma proteins representing no more than 4% of the total proteins present. The iron-binding properties of transferrin and the iron oxidising properties of caeruloplasmin, at a reaction dilution of 1:50, offer considerable protection against organic and inorganic oxygen radicals generated by iron and ascorbate. Plasma thiol group-containing molecules, at concentrations well below those that would be required to compete with the detector molecule (based on known second order rate constants for reaction with hydroxyl radicals) inhibited damage to deoxyribose, but stimulated damage to phospholipids. PMID- 8427874 TI - Recognition of anthracycline binding domains in bovine serum albumin and design of a free fatty acid sensor protein. AB - The mode of binding of N-acylated doxorubicin derivatives to bovine serum albumin (BSA) has been determined by spectrophotometric analysis. A water-insoluble derivative containing a N-hydroxysuccinimide ester moiety on the sugar amino group side-chain is found to react very rapidly with a specific domain (BSAA600) located in the NH2-terminal half of the BSA molecule. A stable covalent protein anthracycline complex with 1:1 molar stoichiometry and containing the albumin monomer is formed. This specific association between albumin and doxorubicin derivative is accompanied by large changes in the spectral characteristics of the anthracycline chromophore. A new strong absorption band at 600 nm is associated to ionization of the chromophore phenolic groups. Titration experiments indicate that the pKa of the protein bound anthracycline is about 3 pH units lower than the pKa of free doxorubicin in aqueous buffer indicating chromophore localization in a basic microenvironment on the albumin molecule. For N-acylated doxorubicin derivatives which associate non-covalently to the BSAA600 domain, the strength of binding is found to be controlled by ionic as well as hydrophobic protein anthracycline interactions. Water-soluble derivatives containing a side-chain carboxylic group bind with Kd approximately 10 microM, which is at least 100-fold more strongly than doxorubicin. The anthracycline chromophore is displaced from the BSAA600 domain in a non-competitive manner by fatty acids ranging in chain length from C6 to C18 and at a fatty acid/BSA molar ratio < 2. We therefore propose a model for the anthracycline binding domain in which the chromophore resides near the opening of the hydrophobic channel into which the fatty acid hydrocarbon chain is inserted. The clusters of basic amino acid residues located at this site may form the basic anthracycline microenvironment. Our results demonstrate that doxorubicin derivatives with a sugar amino group side-chain are well suited as probes for investigations on protein-anthracycline interactions. The practical application of the covalent BSA-DOX complex as a free fatty acid sensor protein for detection of enzymatic release of fatty acids in liposomal and cell membranes is suggested. PMID- 8427875 TI - Identity of D-3-aminoisobutyrate-pyruvate aminotransferase with alanine glyoxylate aminotransferase 2. AB - D-3-Aminoisobutyrate-pyruvate aminotransferase (EC 2.6.1.40) and alanine glyoxylate aminotransferase 2 (EC 2.6.1.44) were co-purified from rat liver as a single protein. The ratio of the two activities remained constant after Sephacryl S-200 chromatography and chromatofocussing. The Km value for beta-alanine as a substrate with 1 mM glyloxylate as amino group acceptor was 1.4 mM. The activity was inhibited by (S)-alanine with Ki = 2.2 mM. The Km for (S)-alanine as substrate with 1 mM glyoxylate as amino group was 6 mM. This activity was inhibited competitively by beta-alanine with Ki = 0.7 mM. (R)-3-aminoisobutyric acid, 5-aminolevulinic acid, NG,NG'-dimethyl-(S)-arginine, and (S)-2-aminobutyric acid were active competitively with respect to beta-alanine with Km of 0.12 mM, 2.1 mM, 6.4 mM and 11.3 mM, respectively. Antiserum to rat liver D-3 aminoisobutyrate-pyruvate aminotransferase inhibited alanine-glyoxylate aminotransferase activity in rat liver in the same way as that of D-3 aminoisobutyrate-pyruvate aminotransferase. Alanine-glyoxylate aminotransferase activity and D-3-aminoisobutyrate-pyruvate aminotransferase activities were inactivated competitively with respect to beta-alanine by 5-fluorouracil and 6 azauracil, which are chemotherapeutic reagents used to cancer. These experiments indicate that D-3-aminoisobutyrate-pyruvate aminotransferase is identical with alanine-glyoxylate aminotransferase 2, aminolevulinate aminotransferase, 2 aminobutyrate aminotransferase and dimetylarginine-pyruvate aminotransferase. PMID- 8427876 TI - Thermostable beta-glucosidase and beta-xylosidase from Thermotoga sp. strain FjSS3-B.1. AB - A beta-D-glucosidase and a beta-D-xylosidase were purified to homogeneity from the thermophilic eubacterium Thermotoga sp. strain FjSS3-B.1. Both enzymes were largely cell-associated and were probably associated with the 'toga' structures of this organism. Using SDS-PAGE they were found to have M(r) values of 75,000 and 92,000, respectively. The beta-glucosidase was active against cellobiose, sophorose and gentiobiose with Km values of 59 mM, 2.7 mM and 6 mM, respectively. The beta-xylosidase had a Km of 2 mM for xylobiose, showed strong activity against p-nitrophenyl alpha-L-arabinofuranoside and p-nitrophenyl alpha-L arabinopyranoside, but was subject to strong substrate inhibition by p nitrophenyl beta-D-xylopyranoside. Both enzymes were extremely thermostable, with half-lives of several hours at 98 degrees C. The thermostabilities of both enzymes were increased further by the addition of either trehalose or betaine. PMID- 8427877 TI - Purification and characterization of two distinct lipases from Candida cylindracea. AB - We have purified and characterized two isoenzymes from a commercial lipase preparation of Candida cylindracea. The purification procedure includes ethanol precipitation and DEAE-Sephacel and Sephacryl HR 100 chromatographies. Lipase A and lipase B were purified 11-fold with a 5% and 21% recovery in activity, respectively. The enzymes have similar amino acid content, N-terminal sequence and molecular weight, but differ on neutral sugar content, hydrophobicity, presence of isoforms and stability to pH and temperature. They also show some differences in the substrate specificity. PMID- 8427878 TI - Purification and characterization of the ganglioside-binding fragment of Clostridium botulinum type E neurotoxin. AB - A way of fragmentation of Clostridium botulinum neurotoxin was carried out to elucidate the structure-function relationship of neurotoxin. The hitherto only plausible fragment was isolated from the trypsin-treated heavy chain of botulinum type E neurotoxin. In the presence of 4 M urea, one protein peak emerged from QAE Sephadex column loaded with the heavy chain mildly treated with trypsin by elution with 0.1 M sodium chloride. Although many protein bands were detected in SDS-PAGE of the treated heavy chain, the eluted protein migrated in a single band to the position of 41,000 Da. The recovery of the 41,000-Da fragment was 28.6%, but with a 2 M urea-containing buffer as eluant, the recovery was less than 12%. The 41,000-Da fragment bound to gangliosides GD1a, GT1b, and GQ1b, to which neurotoxin and the heavy chain bound. The 41,000-Da fragment partially interfered with the binding of 125I-labeled neurotoxin to mouse brain synaptosomes. We have proposed a three-fragment structure (L.H-1.H-2) for botulinum type E neurotoxin. The characters of the 41,000-Da fragment described in this paper seem to substantiated our proposal that type E neurotoxin consists of three fragments, L.H-1.H-2, and that the ganglioside-binding fragment is H-2. PMID- 8427879 TI - The alpha- and beta-subunits of the jacalins are cleavage products from a 17-kDa precursor. AB - The jacalins of three Artocarpus species were purified by affinity chromatography on a desialylated mucin-CNBr-Sepharose 4B column. The beta-chains and the 14 kDa alpha-chains were separated by high pressure liquid chromatography and the 17 kDa chains by preparative electrophoresis. The 17 kDa and 14 kDa chains had a similar highly conserved N-terminal sequence. The beta-chains were different for the three species and Artocarpus champeden contained two different beta-chains. CNBr cleavage of the 17 kDa polypeptide of Artocarpus tonkinensis yielded one peptide more than the 14 kDa. The N-terminal sequence of this fragment was similar to that of the beta-chain proving that this chain results from a proteolytic cleavage at the C-terminus of the 17 kDa peptide. The large heterogeneity of the beta-chains of jacalins from different species could be used as a marker for evolutionary studies on the Artocarpus family. PMID- 8427880 TI - Adaptive changes of intestinal cellular retinol-binding protein, type II following jejunum-bypass operation in the rat. AB - To examine whether the amount of intestinal cellular retinol-binding protein, type II (CRBP(II)) exhibits an adaptive change in a shortened jejunum, the distal end of 5-cm proximal-jejunal segment of 2-mth-old rats was joined to the proximal end of the ileum by an end-to-end anastomosis (jejunum-bypass operation). Three weeks after the operation, the amounts of CRBP(II) in proximal jejunum, proximal ileum and distal ileum were determined using a monospecific antiserum by enzyme linked immunosorbent assay. The jejunum-bypass operation led to a 2-fold increase in the total amounts of CRBP(II) in proximal ileum where hyperplasia occurred; this elevation was in parallel to the increase of total proteins and DNA contents. in the proximal jejunum preceding the bypassed segment, the CRBP(II) level per unit DNA increased by 80%. The jejunum-bypass operation changed neither hepatic total retinol content, nor serum concentrations of retinol and retinol binding protein as compared with the sham-operation, suggesting that retinol absorption was elevated in the shortened small intestine. These results suggest that the increase in CRBP(II) content following jejunum-bypass might be an adaptive response to an enhancement of retinol absorption. PMID- 8427881 TI - Neonatal rat liver contains low concentrations of Cu-Zn superoxide dismutase protein and mRNA. AB - Copper (Cu) distribution among organs and among individual proteins can differ sharply between neonates and adults. The present study found low mRNA and protein levels for neonate rat liver cytosolic Cu-zinc (Zn) superoxide dismutase (SOD), normally among the largest single copper pools in adult rats. Thus, regulation of Cu-Zn SOD mRNA levels contribute to life stage differences in copper distribution. PMID- 8427882 TI - Antioxidant role of dehydroascorbic acid reductase in insects. AB - Dehydroascorbic acid reductase, which catalyses the regeneration of ascorbic acid from dehydroascorbic acid, is reported here to occur widely among insects. Due to the reported absence of glutathione peroxidase in insects and the generally low affinity of catalase for hydrogen peroxide, dehydroascorbic acid reductase may play a pivotal role in the elimination of hydrogen peroxide in insects. PMID- 8427883 TI - Surgical therapy for endocrine tumors of abdominal origin. AB - Surgery continues to be the primary treatment of endocrine neoplasia. Only occasionally are other therapies appropriate and, in those cases, usually only for palliation. In this review, we address the surgical management of endocrine tumors of the pancreas, the adrenal glands, and the gastrointestinal tract (specifically carcinoid tumors). Endocrine tumors, although rare, often present in quite dramatic fashion but are generally associated with excellent survival rates when treated with appropriate surgical resection. PMID- 8427884 TI - Development of resistance mechanisms to the growth-inhibitory effects of transforming growth factor-beta during tumor progression. AB - Members of the transforming growth factor-beta family, especially transforming growth factor-beta type 1, are among the most potent growth-inhibitory factors for epithelial, lymphohematopoietic, and neuroectodermal cells. Resistance to transforming growth factor-beta-mediated growth inhibition is frequently observed in cancers derived from these cell types. We review two important aspects of cancer cell resistance to transforming growth factor-beta: 1) It occurs progressively during the multistep process of tumor progression, and 2) it may encompass a potentially wide range of mechanisms involving transforming growth factor-beta-receptor alterations and cell-signaling defects. Stepwise increases in resistance to the growth-inhibitory action of transforming growth factor-beta may therefore sometimes be attributed to not one but several defects that are involved in transforming growth factor-beta activation, binding, and signaling. These factors accumulate within expanding tumor subclones during disease progression. PMID- 8427885 TI - Preclinical aspects and therapeutic perspectives of acute and chronic leukemias. AB - Increasing understanding of the processes regulating cell growth, differentiation, and the process of malignant transformation as well as the identification of immunologic defense mechanisms against leukemic cells and a better knowledge of the pharmacokinetics of cytostatic drugs, provide the basis for novel therapeutic approaches being explored in preclinical investigations and early clinical trials. These approaches include antileukemic therapy at the molecular level by attempts to block the translation of central oncogenes by antisense oligonucleotides or the interference with abnormal signal transduction by modulation of key enzyme systems, eg, tyrosine kinases or protein kinase C. Reversion of the neoplastic phenotype may be obtained by transfection of tumor cells with wild-type tumor-suppressor genes or restoration of cellular differentiation, ie, by retinoic acid in acute promyelocytic leukemia. Antisense oligonucleotides may also be applied to abrogate abnormal expression of hematopoietic growth factors. Growth factors are also being used clinically to sensitize leukemic cells prior to cytostatic treatment or for a more rapid recovery of normal hematopoiesis after intensive chemotherapy. Immunologic approaches against leukemic cells are based on the identification of disease specific markers, providing the target for antibody therapy with monoclonal antibodies coupled to immunotoxins or allowing the augmentation of cell-mediated immunity by cytokine stimulation, ie, with interleukin-2 or retroviral transfer of genes, eg, the tumor necrosis factor-alpha gene into effector cells. New perspectives also arise from a more detailed knowledge of cytostatic drug pharmacokinetics and the detection of metabolic differences between leukemic cells and their normal hematopoietic counterparts, allowing the design of more specific and tumor-directed drug scheduling. This review highlights some examples of recent preclinical findings that may translate into a better understanding of leukemia biology and ultimately improved therapy for these diseases. PMID- 8427886 TI - Molecular epidemiology of therapy-related leukemias. AB - Over the past 5 years, the field of molecular epidemiology has come of age. Identifying environmental carcinogens and developing methods of dosimetry measurements, which are linked to measures of DNA damage, oncogene activation, and cancer incidence, have opened new avenues of research into cancer causation and prevention. This review summarizes elements of this exciting field, focuses on studies on the etiology of therapy-related leukemias, and suggests potential strategies for its prevention. PMID- 8427887 TI - Leukemia. PMID- 8427888 TI - Endocrine tumors. PMID- 8427889 TI - Cancer biology. PMID- 8427890 TI - Epidemiology and etiology of leukemia. AB - New clinical and epidemiologic studies provide information about the possible causes of human leukemia. Evidence for a viral etiology continues to appear, and the relationship between myelodysplastic syndrome and the leukemias is now linked through molecular genetic studies. Molecular mechanisms of leukemogenesis are being understood through evaluation of preleukemic conditions and predisposing medical illnesses. Epipodophyllotoxins and, to a lesser extent, cisplatin are being linked causally to secondary leukemia. Potential environmental causes of leukemia are being intensively investigated with both positive and negative results. The literature on the epidemiology of leukemia is growing rapidly, and important leads toward a more complete understanding of its etiology are emerging. PMID- 8427891 TI - Implications of detection of minimal residual disease. AB - Complete remission can be achieved in the majority of children and adults with acute leukemia, but relapses are frequent due to persistent disease below the level of detectability of light microscopy. Recently, more sensitive molecular biologic and immunologic approaches have been developed for monitoring of leukemia-specific or -associated markers. Methods are now available for follow-up studies in all leukemic entities. New insights have already been gained into the efficiency of therapeutic strategies; persistence of a leukemic marker is a common finding in patients shortly after induction treatment and does not necessarily predict relapse, and increase of positive signals or reappearance of a leukemic marker in complete remission often precedes relapse. However, not all methods are standardized, and levels of sensitivity differ. Conclusions for stratification of postremission therapy can only be drawn after evaluation of ongoing, prospective studies. PMID- 8427892 TI - Chronic lymphoproliferative disorders: chronic lymphocytic leukemia and hairy cell leukemia. AB - A consistent stimulus to increase interest in malignant disease is the discovery of new successful treatments. The dramatic activity of the nucleoside analogues, 2'-deoxycoformycin, fludarabine, and 2-chlorodeoxyadenosine, in indolent lymphoproliferative diseases has caused a reawakening of interest in the biology and treatment of chronic lymphocytic leukemia and hairy-cell leukemia. The high response rate has led to a reevaluation of criteria for response in these diseases as well as indications for treatment and prognostic factors. PMID- 8427893 TI - Therapy for childhood acute lymphoblastic leukemia. AB - Recent advances in therapy for childhood acute lymphoblastic leukemia have come primarily from the intensive laboratory study of patient's lymphoblasts. DNA ploidy determination, the analysis of specific chromosomal translocations, and in vitro chemosensitivity studies now facilitate the stratification of risk groups and the prediction of treatment outcome. More is known about the heterogeneity of molecular defects involved in leukemogenesis, and this information is being exploited to devise sensitive tests for minimal residual disease. Conventional chemotherapy of childhood acute lymphoblastic leukemia is associated with adverse neuropsychological sequelae and second malignancies when intensive epipodophyllotoxin therapy is used. Treatment of relapsed acute lymphoblastic leukemia remains problematic. The development of alternative donor marrow sources will expand the role of bone marrow transplantation, which has provided better outcomes for a limited number of patients. We are still waiting to spawn novel therapeutic agents that are more effective and less toxic than present chemotherapy. PMID- 8427894 TI - Aspects of treatment of acute myeloid leukemia in adults. AB - The treatment of acute myeloid leukemia has made steady rather than spectacular progress in the past few years, principally because of improved supportive care. The use of hematopoietic growth factors may contribute to the speed of normal bone marrow recovery following treatment; however, their potential role as modifiers of drug pharmacokinetics may be equally valuable in overcoming drug resistance. The full range of expression of the phenomenon of resistance is being intensively explored, and therapeutic opportunities for overcoming the P glycoprotein pump are now being introduced. Gene therapy and exciting immunologic manipulations are also developing rapidly, and the role of suppressor genes has reached a fascinating point in research and clinical applications. PMID- 8427895 TI - Molecular genetics of neuroendocrine tumors. AB - Neuroendocrine tumors are a group of malignancies that arise from primitive neuroectodermal cells. There is, however, considerable variation in the location and clinical behavior of these tumors. They are often difficult to distinguish from other malignancies and difficult to classify. Recent advances in molecular genetics have attempted to resolve some issues regarding accurate diagnosis, classification into prognostic subgroups, and understanding of basic common mechanisms underlying the development of these tumors. Correlation of morphologic changes with molecular changes induced by some potential therapeutic differentiating agents may lead to more rational therapy for these malignancies. PMID- 8427896 TI - Computed tomographic, magnetic resonance, and monoclonal antibody imaging of endocrine tumors. AB - Recent advances in diagnostic imaging with computed tomography, magnetic resonance imaging, and monoclonal antibody imaging in endocrine tumor evaluation are reviewed. Computed tomography is a mature technology whose role has not changed recently. Magnetic resonance imaging is continuing to advance in scan techniques, scan time reduction, and use of intravenous contrast agents; its future role on a routine basis may allow it to supplant computed tomography in endocrine tumor identification. Monoclonal antibody imaging has not yet proven to be as reliably diagnostic for tumor identification as is computed tomography, but continuing technical advances predict an advancing role for diagnosing endocrine tumors with this modality. PMID- 8427897 TI - Whatever happened to HIV dementia? PMID- 8427898 TI - Factors associated with trichomoniasis, candidiasis and bacterial vaginosis. AB - Of 6125 women attending an STD clinic from 1988 to 1991, 5365 (88%) were tested for vaginitis of whom 97 (1.8%) had trichomoniasis, 945 (17.6%) had candidiasis, 734 (13.7%) had bacterial vaginosis and 3628 (67.6%) were free of vaginal infection. Dual infections occurred in 49 (0.9%) patients. Independent predictors for trichomoniasis by multivariate analysis were being pregnant (odds ratio (OR) = 2.4), having vaginal discharge or dysuria (OR = 4.7), being Aboriginal (OR = 4.3), being Asian (OR = 5.0), being unemployed (OR = 2.1) or tattoed (OR = 1.9). Many factors, including use of oral contraception (OR = 1.2) and current antibiotic medication (OR = 1.5), had a small significant association with candidiasis. Independent predictors for bacterial vaginosis were having multiple sex partners in the past month (OR = 1.6), being unmarried (OR = 1.5), being unemployed (OR = 1.3) being a prostitute (OR = 1.5) and not currently using antibiotic medication (OR = 2.5). The epidemiological profiles were consistent with trichomoniasis and bacterial vaginosis being sexually transmitted diseases with epidemiology different from that of gonorrhoea and chlamydia and different from each other, and candidiasis being a disease in which constitutional factors are more important than issues relating to sexual transmission. PMID- 8427899 TI - Partner notification for human immunodeficiency virus infection in Colorado: results across index case groups and costs. AB - To evaluate human immunodeficiency virus (HIV) partner notification and referral across index case groups, we analysed results of services provided by the Colorado Department of Health during 1988. Services were offered to 231 index cases; 226 (98%) accepted; 124 (65%) with unsafe behaviours identified 239 partners; 79% of partners were notified; 68% accepted counselling. Seropositivity was 21% in newly tested partners. Index cases chose patient referral for 25% of partners and referred 20% of eligible partners; the provider referred 71%. Index case acceptance of service, proportion of index cases with newly identified HIV positive partners, and choice of partner referral method were similar across groups. Gay men identified fewer partners, had a lower proportion of partners accepting new counselling and testing and referred a lower proportion of partners than heterosexual men. The total costs of the service were $19,496. Twelve new cases of HIV were identified through patient and provider referral and cost per case identified was $1625. Partner notification and provider referral should be offered to all HIV infected individuals in Colorado, as few differences across groups emerged and only 20% of located and eligible partners received counselling through patient referral. PMID- 8427900 TI - Antibiotic susceptibility, serovars and auxotypes of gonococcal isolates in Stockholm. Relation to geographical origin of the infection. AB - The antibiotic susceptibility, serovars and auxotypes were investigated in gonococcal strains isolated from all patients with gonorrhoea during one year in Stockholm, Sweden. The results were correlated to geographical origin of the infection. A total of 394 gonococcal strains were isolated from 392 patients, 135 (34%) women and 257 (66%) men. Beta-lactamase-producing gonococcal strains (PPNG) were isolated from 5% of the women and 16% of the men. Men had acquired their infection abroad more often than women (54% vs 33%) (P < 0.001). The majority (81%) of the PPNG infections were imported. Some serovars and auxotypes were more common among imported strains than among indigenous ones. All strains were sensitive to spectinomycin and 2 strains had decreased susceptibility to norfloxacin and ciprofloxacin. Decreased susceptibility to benzylpenicillin, ampicillin, doxycycline and cefuroxime was related to the geographical origin of the strains with strains imported from regions other than Europe being the most resistant. PMID- 8427901 TI - Management of cervical intraepithelial neoplasia in departments of genitourinary medicine--a national survey. AB - A national survey of the methods used in the diagnosis and treatment of cervical intraepithelial neoplasia (CIN) in the departments of genitourinary medicine (GUM) in the United Kingdom had been conducted through a postal questionnaire. The findings indicate that at present of 157 responders to the questionnaire, 43 (27.3%) take an active part in the management of CIN. The different modes of management and follow-up are presented. PMID- 8427902 TI - Detection of IgG, IgM and IgA antibodies in patients with uncomplicated Chlamydia trachomatis infection: a comparison between enzyme linked immunofluorescent assay and isolation in cell culture. AB - The diagnostic value of serum IgG, IgM and IgA in patients with uncomplicated urogenital Chlamydia trachomatis infection was compared with isolation in cell culture. C. trachomatis specific antibodies were determined with an enzyme linked immunofluorescent assay using elementary bodies from C. trachomatis serotypes E,F,H,I,J and LGV2 as antigens. At least two sera from each patient were tested and cultures were also established on the same day. Excluding the IgM titres in men, significantly more IgG, IgA and IgM and combinations of these antibodies were observed in culture positive patients. The sensitivity with which IgG titres in men or IgG and/or IgM titres in men and women could be determined, was significantly lower using C. trachomatis LGV2 as the only antigen than when all 6 antigens were used. The presence of 10 or more leucocytes in the urine sediment of men correlated positively with an IgG or an IgG and/or IgM titre. PMID- 8427903 TI - Beta-2 microglobulin as a marker of HIV disease status in Nairobi, Kenya. AB - Serum beta-2 microglobulin (beta 2-M) has prognostic value similar to lymphocyte profiles for predicting disease progression in those infected with the human immunodeficiency virus (HIV). However, the relationship between beta 2-M and HIV disease progression among inhabitants of countries with endemic tropical diseases has not been evaluated. To determine the relationship between serum beta 2-M levels and HIV infection and disease status in an African population, serum beta 2-M levels were measured in 369 patients attending a sexually transmitted disease (STD) clinic in Nairobi, Kenya. Mean serum beta 2-M was significantly higher in HIV seropositive than in HIV seronegative individuals. Among HIV infected patients, higher mean beta 2-M levels were observed in those with HIV associated symptoms or laboratory markers of advanced HIV disease. Significant inverse correlations between beta 2-M and the percentage of CD4 lymphocytes or CD4/CD8 ratio were found. These findings suggest that beta 2-M measurements may have prognostic value for HIV infected populations in developing countries. PMID- 8427904 TI - The value of tests of cure following cervical chlamydial infection. AB - Test of cure (TOC) was performed 2, 4 and 6 weeks after treatment for cervical chlamydia infection with 10-14 days of Deteclo one tablet twice daily, erythromycin 500 mg twice daily or doxycycline 100 mg twice daily. Testing was by chlamydia culture and IDEIA (DAKO diagnostics Ltd). Discrepant results were subsequently checked by immunofluorescence (Syva MicroTrak) of both sets of left over transport media. Two hundred and three patients attended on at least one occasion; 189, 146 and 107 at 2, 4 and 6 weeks respectively. Of these 127, 70 and 34, respectively, denied sexual intercourse or had consistently used condoms. Fourteen were positive over the study period by either or both methods of detection. Of 8 culture positive results 3 were negative by IDEIA. Two of these had elementary bodies (EBs) on immunofluorescence of both sets of saved transport media. One had EBs on immunofluorescence of the saved culture transport medium only. None of the 6 IDEIA positive, culture negative patients had immunofluorescent EBs in the IDEIA transport media although one had EBs in the saved culture transport medium. One IDEIA suspicious, culture negative patient had EBs in both sets of saved transport media. There was no significant difference in the rate of chlamydia detection from patients admitting to or denying unprotected intercourse. TOC has a low yield in cases of cervical chlamydial infection when there has been careful contact tracing and treatment has been completed. If TOC is performed culture should be used if available and where antigen detection methods are used confirmation should be sought for any positive results. PMID- 8427905 TI - Vulval Crohn's disease mimicking genital herpes. PMID- 8427906 TI - STD in travellers presenting to a GUM clinic. PMID- 8427907 TI - Do we need vulvar clinics in genitourinary medicine departments? PMID- 8427908 TI - Current smoking habits and genital infections in women. PMID- 8427909 TI - Herpes simplex virus infection in pregnancy and its management. PMID- 8427910 TI - Anogenital non-gonococcal neisseriae: prevalence and clinical significance. AB - Over a 13-year period non-gonococcal neisseriae (NGN) were isolated from 114 of 88,670 patients (0.13%) screened for anogenital gonorrhoea at a Genitourinary Medicine Unit. During the same period there were approximately 9000 anogenital gonococcal infections (10%). The prevalence of NGN was 0.09% (27/31,500) in women, 0.04% (20/52,800) in heterosexual men and 1.5% (67/4370) in homosexual men: the differences in prevalence between women and heterosexual men (P < 0.01) and between heterosexual patients and homosexual men (P < 0.001) are highly significant. Neisseria meningitidis was isolated most frequently and accounted for 85% (99/114) of the NGN. Whenever possible, N. meningitidis was serogrouped and its occurrence correlated with patient symptoms. Eleven of 18 heterosexual men who had meningococci isolated from their urethras had urethritis but co existing chlamydial infection was excluded in only 5. None of 9 women with cervical colonization had clinical evidence of pelvic inflammation. Only one of 49 men with rectal colonization had proctitis. The management of anogenital NGN infection is discussed in relation to our findings and those of previously published studies. Within each patient group the prevalence and incidence of anogenital NGN were similar at the beginning and end of the study period indicating that levels have not been influenced by the advent of AIDS. PMID- 8427911 TI - [Health plans and prevention in primary care]. PMID- 8427912 TI - [A comparative analysis of 11 adult case histories used in primary care]. AB - OBJECTIVE: A comparative analysis of the different models of adult case history at primary care (PC) level available in the Spanish state. Their degree of homogeneity, both in design and information-gathering capacity, was evaluated. DESIGN: A descriptive study. MATERIAL: Eleven individual adult PC cases histories, still open in 1991. MAIN MEASUREMENTS AND RESULTS: Only four of the models allowed more than 75% of the basic data to be gathered. Three histories had up to six loose pages. In six, the presence of specific records was verified. CONCLUSIONS: There is a lack of standardisation both in the designs of case histories and in the information which these allow to be gathered. Information was scattered because of the presence of specific records and of loose pages. Systematic inclusion of preventive activities into the records is not sufficiently normalised. PMID- 8427913 TI - [An evaluation of the information for the patient in pharmacies]. AB - OBJECTIVE: To study the nature of the information that is supplied by pharmacies in Bizkaia to their patients and the factors impinging on this information. DESIGN: Crossover study of a sample of users of pharmacies. SITE. The research was done in 10 representative community pharmacies in the province of Bizkaia, included in which were rural and urban pharmacies. PATIENTS: 19,875 patients who used these pharmacies during the month of the study (May, 1991). INTERVENTIONS: The information given the patient by the pharmacist was analyzed, as to whether the request came from the patient him/herself or not and as to the subject matter of this information. MAIN MEASUREMENTS AND RESULTS: The pharmacist gave information to 4,162 of the 19,875 people observed, i.e. 1 in every 5 patients. More information was exchanged as a result of patients' request (69.17% of all the information given) than the pharmacist's volunteering it (30.38%). These figures constituted 14.48% and 6.45% respectively of the total number of people using the pharmacies during this month. The greatest number of requests for information by patients was concerning illnesses and their treatment: 22.4% of all the information. CONCLUSIONS: Information is given in pharmacies concerning health questions. However a mechanism needs to be found so that this information is of sufficient scope to contribute towards greater efficiency of treatment and health programmes. PMID- 8427914 TI - [The consumption of alcoholic drinks in adolescence]. AB - OBJECTIVE: To identify the factors involved in the use and abuse of alcoholic drinks among a group of adolescents attending school. DESIGN: A descriptive, crossover study. SITE. 23 schools in the city of Barcelona. PARTICIPANTS: Students doing the sixth and seventh grades of EGB (in general, between 11 and 13 years old). MAIN MEASUREMENTS AND RESULTS: An in-school questionnaire, which gathered data from 2,033 students (92.2% coverage), was carried out. The students' replies were used to construct one index of probable alcohol abuse (IPAA) and another of the penetration of alcohol in the environment (IPAE). 51.9% had consumed some alcoholic drink, with 9.5 as the average age for the first drink. 1.3% drank alcohol every day. 1.8% said they had been drunk two or more times during the previous six months. 3.1% had bought alcohol for their own use. 8% of the students had at least one point on the IPAA. The seventh-grade students scored higher on the IPAE. The alcohol abuse indicators increased if the parents consumed alcohol, but more so if their peers were also consumers (older siblings or friends). There is a link between alcohol and tobacco consumption. CONCLUSIONS: The data showed how the increased penetration of alcohol into schoolchildren's environment resulted in abusive consumption being more likely. The importance of preventing the abuse of alcoholic drinks among adolescents was confirmed. PMID- 8427915 TI - [Thyroid function tests and their use by the general practitioner]. AB - OBJECTIVE: We aimed to discover how thyroid function tests (TFTs) were used in general practice (GP) consultations. We would evaluate how many TFTs were requested, their efficiency, cost and their impact on referrals to endocrinology. DESIGN: This was a longitudinal, retrospective and observational study. SITE. At the Primary Care level, in the La Chana Health Centre, Granada. PATIENTS: 69 patients with possible Thyroid Disease, and who had a TFT performed by their GP, were surveyed. The test comprised parallel TSH, free T4 and free T3. MAIN MEASUREMENTS AND RESULTS: 2.46 TFTs per GP per year were requested. 14 showed positive (20.28%) and a definite diagnosis of Thyroid Function Disease (TFD) was made in 11 cases (15.94%). The cost per test was 1,718 pesetas; and per diagnosis, 8,467 pesetas. 78.26% of the cases were resolved by the GP without need of referral. CONCLUSIONS: The GPs carried out few FTFs; but those they did were very efficient. Cost was low and patients benefited from the fact that their GPs were able to use this technique. Endocrinology referrals were reasonable. All general practitioners should have use of TFTs. PMID- 8427916 TI - [The perception by the population of the functions and components of the primary care team]. AB - OBJECTIVE: To find out how the population in Health Area IV of Navarra (Tafalla) see the new components of the primary care team (PCT) and their health education functions. DESIGN: A crossover descriptive study. SITE. Health Area IV of Navarra (Tafalla). PARTICIPANTS: 816 people aged 18 or over and who lived in the area under study made up a sample stratified according to age, gender and where they lived. Interviewees were selected through the random routes method. MAIN MEASUREMENTS AND RESULTS: A personal interview with a questionnaire was carried out in the home of the participants during March and April, 1988. There was widespread ignorance of what the PCT meant; paradoxically, this ignorance was significantly higher (p = 0.014) in the base areas where EAP had been set up (80%) than in those areas where it had not (73%). In spite of this, in the Tafalla base area the components of the PCT had begun to be understood. Only 7% replied that they knew other functions, apart from health care, of the PCTs: here, it was in the areas where PCTs had been set up that there was significantly greater knowledge (p = 0.0004). CONCLUSIONS: A large percentage of the population is ignorant regarding new elements (staff and functions) introduced into the PCT. PMID- 8427917 TI - [A study on anti-infective agent self-medication in 2 pharmacy offices]. AB - OBJECTIVE: To describe and analyse the demand for unprescribed drugs against infections in two pharmacy departments, as a first step towards constructing closer cooperation between primary care physicians, chemists from the pharmacy service and chemists from a pharmacy department. DESIGN: A descriptive and prospective study based on observation. SITE. This study was carried out at two pharmacy departments: the Embajadores Health Centre and the pharmacy service of the hospital to which patients were referred from Embajadores. PATIENTS AND PARTICIPANTS: The target population were all those patients who requested a drug to combat infection in the pharmacy departments during the six months of the study (July to December, 1991). MAIN MEASUREMENTS AND RESULTS: A total of 186 requests without a prescription for drugs to combat infection was recorded. The profile of those so requesting was of an almost equal number for men and women between 31 and 50 years old. The main reasons behind the requests were throat, dental and urinary-genital infections. The consumption of drug per category of illness was, from greater to lesser, penicillin, sulfamides, urinary chemotherapies, tetracyclines, macrolides and cephalosporins. CONCLUSIONS: Although there appears to be consistency between the drug infection requested and the reason for requesting it, some health education activities aimed at the general population need to be organised, in order to raise awareness of the use and abuse of antibiotics. PMID- 8427918 TI - [The detection of and approach to growth retardation in primary care]. PMID- 8427919 TI - [Community participation and health: maintaining the sense of proportion]. PMID- 8427920 TI - [The future leading role of primary care in HIV/AIDS]. PMID- 8427921 TI - [The profile of the triazolam consumer]. PMID- 8427922 TI - [Domiciliary parenteral nutrition]. PMID- 8427923 TI - [The incorrect use of the GHQ]. PMID- 8427924 TI - [Accidental paraquat poisoning]. PMID- 8427925 TI - [An analysis of research projects on primary care presented to the Fondo de Investigacion Sanitaria during 1991]. AB - DESIGN: Observational cross-sectional study. SETTING: Proposals on primary health care (domain, functions and activities) identified among a total of the 1412, belonging to all topics, submitted to the Fondo de Investigacion Sanitaria during the 1991 exercise. INTERVENTIONS: Proposals were classified according the degree of quality in the methodological fulfillment, pointed out those accepted for grant by the Technical Evaluation Committees. MEASUREMENTS AND RESULTS: 55 proposals about Primary Health Care were identified which account for 3.8% of the all proposals submitted. The degree of topic pertinence was very high with hypothesis and objectives well set out. There was a poor fulfillment of the methods and plan of action. Those projects with a very good fulfillment were accepted in a 90% of the cases. CONCLUSIONS: The reviewed proposals shaw a good pertinence in the research on primary health care. We could notice a growing portion of researchers well trained with exigent methodological criteria. However, it remains a majority of proposals with poor methodological fulfillment which could easily be improved by increasing research training and expert support. Some advices are offered for the elaboration of research projects. PMID- 8427926 TI - Solution structure of peptides containing two dehydro-phenylalanine residues: a CD investigation. AB - The peptides Ac-delta Phe-Ala-delta Phe-NH-Me (1), Ac-delta Phe-Val-delta Phe-NH Me (2), Ac-delta Phe-Gly-delta Phe-Ala-OMe (3), and Boc-Ala-delta Phe-Gly-delta Phe-Ala-OMe (4), containing two dehydro-phenylalanine (delta Phe) residues, were synthesized and the solution structure investigated in various solvents. The nmr and CD measurements indicate that all the dehydropeptides examined adopt 3(10) helical conformations in solution. The tripeptides 1 and 2 exhibited an intense negative CD exciton couplet, which was assigned to the right-handed screw sense, while the tetrapeptide 3 displayed a CD couplet having opposite sign, which was assigned to the left-handed helical sense. In the pentapeptide 4 the sense of the helix was found to vary with solvent and temperature, as demonstrated by the sign reversal of the CD spectrum. The right-handed sense dominates in hexafluoro-2 propanol, whereas a left-handed helix prevails in chloroform, acetonitrile and methanol. A crucial role for this behavior is likely to be played by the two alanine residues positioned respectively at the head and tail of the sequence, which favor conformations having opposite screw senses. PMID- 8427927 TI - Hydrational and intrinsic compressibilities of globular proteins. AB - Partial compressibilities of globular proteins in water are reviewed. Contribution of hydrational and of intrinsic compressibilities to experimental partial quantity have been evaluated from ultrasonic data using two independent methods: (a) additive calculation of the hydrational contributions of the surface atomic groups and (b) an analysis of correlation between partial compressibility and molecular surface area. The value (14 +/- 3) X 10(-6) bar(-1) for the isothermal compressibility coefficient of the protein interior at 25 degrees C was obtained as an average value for variety of globular proteins. This value is similar to that of solid organic polymers. Possible relaxation contribution to partial compressibility is roughly estimated from comparison of thermodynamic with x-ray data on protein compressibility. The average compressibility of water in the hydration shell of proteins was found to be 35 X 10(-6) bar(-1), which is 20% less than that of pure water. PMID- 8427928 TI - Volume changes correlate with entropies and enthalpies in the formation of nucleic acid homoduplexes: differential hydration of A and B conformations. AB - We have used a combination of densimetric, calorimetric, and uv absorption techniques to obtain a complete thermodynamic characterization for the formation of nucleic acid homoduplexes of known sequence and conformation. The volume change delta V accompanying the formation of four duplexes was interpreted to reflect changes in hydration based on the electrostriction phenomenon. In 10 mM sodium phosphate buffer at pH 7, the magnitude of the measured delta V's ranged from -2.0 to +7.2 ml/mol base pair and followed the order of poly(rA).poly(dT) approximately poly(dA).poly(dT) < poly(rA).poly(dU) approximately poly(rA).poly(rU). Inclusion of 100 mM NaCl in the same buffer gave the range of 17.4 to -2.3 mL/mol base pair and the following order: poly(dA).poly(dT) < poly(rA).poly(dT) < poly(rA).poly(rU) approximately poly(rA).polyr(dU). Standard thermodynamic profiles of forming these duplexes from their corresponding complementary single strands indicated similar free energies that resulted from the compensation of favorable enthalpies with unfavorable entropies along with a similar counterion uptake at both ionic strengths. The differences in these compensating effects of entropy and enthalpy correlated very well with the volume change measurements in a manner suggesting that the homoduplexes in the B conformation are more hydrated than are those in the A conformation. Moreover, the increased thermal stability of these homoduplexes resulted from an increase in the salt concentration corresponding to larger hydration levels as reflected by the delta V results. PMID- 8427929 TI - Kinetic analysis of the hydrodynamic transition accompanying protein folding using size exclusion chromatography. 1. Denaturant dependent baseline changes. AB - Size exclusion profiles of proteins with persistent conformations exhibit broad asymmetric peaks whose shape and elution times are dependent on denaturant concentration. The collective elution profiles were precisely simulated by an apparent binding model that treats the denaturant dependence in terms of an apparent matrix binding. The model requires three experimentally measurable parameters: the elution time for the unbound protein, an apparent association equilibrium constant for binding, and an apparent exchange time for binding. The denaturant dependence for each of these parameters is related to the accessible surface area of the protein. PMID- 8427931 TI - Microwave absorption spectroscopy of DNA. AB - By utilizing a novel approach to microwave spectrometry, we have measured the absolute absorption spectrum of plasmid DNA (pUC8.c2), in buffered aqueous solution, from 5 to 20 GHz. Our technique does not suffer from the same experimental difficulties that plague other methods. We observe no absorption resonances in this frequency range, but we do see broadband differences, between DNA and pure buffer, that are attributable to changes in the ionic conductivity of the solutions. These results constitute the first verification, by a totally different technique, of the absence of resonances in the microwave absorption spectrum of DNA, and the first data obtained by any technique in the 10-20-GHz band. PMID- 8427930 TI - Kinetic analysis of the hydrodynamic transition accompanying protein folding using size exclusion chromatography. 2. Comparison of spectral and chromatographic kinetic analyses. AB - The kinetics of the hydrodynamic volume change associated with the unfolding and refolding of a globular protein can be observed using high performance size exclusion chromatography. Chromatographic profiles that evidence such dynamics can be simulated using equations in which chromatographic partitioning and the conformational transition are described in terms of a finite difference algorithm incorporating an apparent binding model to generate broad and asymmetric peaks. Application of these equations to the simple two-state unfolding transition of ribonuclease A in guanidine hydrochloride indicates that reliable kinetic parameters can be obtained using these equations. PMID- 8427932 TI - Temperature-induced conformational transition in xanthans with partially hydrolyzed side chains. AB - The conformational properties of xanthans with partially hydrolyzed side chains were investigated by optical rotation, CD, and differential scanning calorimetry (DSC). All variants displayed the well-known temperature-driven, cooperative order-disorder transition, and both optical rotation and DSC showed that the transition temperature was essentially independent of the content of terminal beta-mannose. It was found that up to 80% of the changes in the specific optical rotation accompanying the transition reflects conformational changes linked to the terminal beta-mannose in the side chains. Modification of the side chains also affected the CD when xanthan was in the ordered state, but in this case the data suggest that the glucuronic acid is the major component determining the magnitude of the CD signal. DSC measurements showed that the transition enthalpy (delta Hcal) increased linearly with the fraction of beta-mannose, again indicating that a significant part (up to 80%) of delta Hcal reflects conformational changes in the side chains. The conformational transition of the xanthan variants generally showed a higher degree of cooperativity (sharper transition) than unmodified, pyruvated xanthan. Calculation of the cooperativity parameter sigma by means of the Zimm-Bragg theory (OR data) or from the ratio between delta Hcal and the van't Hoff enthalpy (delta HvH) using DSC data showed a correlation between sigma and the content of beta-mannose, but the two methods gave different results when the content of beta-mannose approached 100%. The ionic strength dependence of the transition temperature, expressed as d (log I)/d(T-1m), was nearly identical for intact xanthan and a sample containing only 6% of the terminal beta-mannose. Application of the Manning polyelectrolyte theory does not readily account for the observed delta Hcal values, neither does it provide new information on the nature of the ordered and disordered conformations in xanthan. PMID- 8427933 TI - Helix-forming tendencies of amino acids depend on their sequence contexts: tripeptides AFG and FAG show incipient beta-bulge formation in their crystal structures. AB - Many of the theoretical methods used for predicting the occurrence of alpha helices in peptides are based on the helical preferences of amino acid monomer residues. In order to check whether the helix-forming tendencies are based on helical preferences of monomers only or also on their sequence contexts, we synthesized permuted sequences of the tripeptides GAF, GAV, and GAL that formed crystalline helices with near alpha-helical conformation. The tripeptides AFG and FAG formed good crystals. The x-ray crystallographic studies of AFG and FAG showed that though they contain the same amino acids as GAF but in different sequences, they do not assume a helical conformation in the solid state. On the other hand, AFG and FAG, which contain the same amino acids but in a different sequence, exhibit nearly the same backbone torsion angles corresponding to an incipient formation of a beta-bulge, and exhibit nearly identical unit cells and crystal structures. Based on these results, it appears that the helix-forming tendencies of amino acids depend on the sequence context in which it occurs in a polypeptide. The synthetic peptides AFG (L-Ala-L-Phe-Gly) and FAG (L-Phe-L-Ala Gly), C14H19N3O4, crystallize in the orthorhombic space group P2(1)2(1)2(1), with a = 5.232(1), b = 14.622(2), c = 19.157(3) A, Dx = 1.329 g cm-3, Z = 4, R = 0.041 for 549 reflections for AFG, and with a = 5.488(2), b = 14.189(1), c = 18.562(1) A, Dx = 1.348 g cm-3, Z = 4, R = 0.038 for 919 reflections for FAG. Unlike the other tripeptides GAF, GGV, GAL, and GAI, the crystals of AFG and FAG do not contain water molecule, and the molecules of AFG aor FAG do not show the helical conformation. The torsion angles at the backbone of the peptide are psi 1 = 144.5(5) degrees; phi 2, psi 2 = -98.1(6) degrees, -65.2(6) degrees; phi 3, psi 13, psi 31 = 154.1(6) degrees, -173.6(6) degrees, 6.9(8) degrees for AFG; and psi 1 = 162.6(3) degrees; phi 2, psi 2 = -96.7(4) degrees, -46.3(4) degrees; phi 3, psi 13, psi 31 = 150.1(3) degrees, -168.7(3) degrees, 12.2(5) degrees for FAG. The conformation angles (phi, psi) for residues 2 and 3 for both AFG and FAG show incipient formation of an beta-bulge. PMID- 8427934 TI - Necessary conditions for avoiding incorrect polypeptide folds in conformational search by energy minimization. AB - Low energy conformations have been generated for melittin, pancreatic polypeptide, and ribonuclease S-peptide, both in the vicinity of x-ray structures by energy refinement and by an unconstrained search over the entire conformational space. Since the structural polymorphism of these medium-sized peptides in crystal and solution is moderate, comparing the calculated conformation to x-ray and nmr data provides information on local and global behavior of potential functions. Local analysis includes standardization calculations, which show that models with standard geometry can approximate good resolution x-ray data with less than 0.5 A rms deviation (RMSD). However, the atomic coordinates are shifted up to 2 A RMSD by local energy minimization, and thus 2 A is generally the smallest RMSD value one can target in a conformational search using the same energy evaluation models. The unconstrained search was performed by a buildup-type method based on dynamic programming. To accelerate the generation of structures in the conformational search, we used the ECEPP potential, defined in terms of standard polypeptide geometry. A number of low energy conformations were further refined by relaxing the assumption of standard bond lengths and bond angles through the use of the CHARMM potential, and the hydrophobic folding energies of Eisenberg and McLachlan were calculated. Each conformation is described in terms of the RMSD from the native, hydrogen-bonding structure, solvent-accessible surface area, and the ratio of surfaces corresponding to nonpolar and polar residues. The unconstrained search finds conformations that are different from the native, sometimes substantially, and in addition, have lower conformational energies than the native. The origin of deviations is different for each of the three peptides, but in all examples the refined x-ray structures have lower energies than the calculated incorrect folds when (1) the assumption of standard bond lengths and bond angles is relaxed; (2) a small and constant effective dielectric permittivity (epsilon < 10) is used; and (3) the hydrophobic folding energy is incorporated into the potential. PMID- 8427935 TI - Macromolecular cyclization of (1-->6)-branched-(1-->3)-beta-D-glucans observed after denaturation-renaturation of the triple-helical structure. PMID- 8427936 TI - NMR studies of an extrahelical cytosine in an A.T rich region of a deoxyribodecanucleotide. AB - One-dimensional and two-dimensional (2D) nmr experiments were carried out on an oligonucleotide duplex that contains an unpaired cytosine, d(GCGAACAAGCG).d (CGCTTTTCGC), which will be referred to as the C-bulge decamer. Evidence from one dimensional nuclear Overhauser effect (NOE) experiments on the exchangeable protons indicates that the unpaired cytosine is extrahelical. This conclusion is also supported by numerous cross-peaks in the 2D NOE spectroscopy (NOESY) spectrum of the nonexchangeable protons. The assignments for all of the resonances, with the exception of the H5' and H5" resonances, have been made through the use of 2D NOESY, correlated spectroscopy (COSY), and relayed COSY experiments. The temperature dependence of the C (H6) resonance chemical shifts indicates that the unpaired cytosine shows unusual behavior compared to other cytosines in the duplex. A comparison of chemical shifts for all the assigned resonances of the duplexes with and without the unpaired cytosine suggests that the majority of the structural perturbation is localized in the A.T tract surrounding the unpaired base. The behavior of the imino resonances as a function of temperature also indicates that the perturbation to the duplex is localized and destabilizes the A.T base pairs adjacent to the unpaired base. PMID- 8427937 TI - Influence of electrostatic interactions on the sugar-phosphate backbone conformation in DNA. AB - The influence of sugar ring flexibility in DNA on the mechanism of the B <===> A conformational transition is studied. The dipole moment of the deoxyribose as a function of its puckered states is calculated by the quantum-mechanical method using the MINDO/3 approximation. The interaction of the sugar dipole with the neighbor molecular groups in polynucleotide chains is estimated. The sugar dipole interaction with phosphate groups and counterions is shown to be strong and capable to deform the pseudorotation potential of deoxyribose. The effective pseudorotation potential of sugar ring in the B and A helices is obtained. The results are used to explain the behavior of Raman bands in the region of sugar phosphate vibrations. The mechanism of the effect of electrostatic forces on the sugar-phosphate backbone conformation, which is essential for the B <===> A and other structure transitions, is offered. PMID- 8427938 TI - Measurement of restricted rotational diffusion of fluorescent lipids in supported planar phospholipid monolayers using angle-dependent polarized fluorescence photobleaching recovery. AB - A theory describing the shapes of polarized fluorescence photobleaching recovery (PFPR) curves for a population of fluorophores undergoing restricted rotational diffusion in two-dimensional systems such as planar membranes has been developed. In this model, restricted rotational diffusion of the fluorophores is described by using reflective boundary conditions, in which the fluorophores are assumed to diffuse freely but only within an angular space of width 2 omega. The magnitude and apparent rate of the PFPR postbleach fluorescence curves are a function of both omega and the angle between the bleaching and observation beam polarizations psi. It is shown that estimates of the degree of rotational restriction omega may be obtained from changes in the psi-dependent postbleach fluorescence intensities. Using angle-dependent PFPR, slow rotational reorientations of the fluorescent lipid analogue 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine in distearoylphosphatidylcholine Langmuir-Blodgett monolayers deposited on octadecyltrichlorosilane-treated fused quartz were measured. As theoretically predicted for a rotationally restricted fluorophore population, both the initial F psi (0) and final F psi (infinity) postbleach fluorescence intensities varied as a function of psi, and no measurable change in the postbleach fluorescence intensities was observed for psi = 45 degrees. Using the theory for restricted rotational motion, the psi-dependent variations of the final fluorescence intensities F psi (infinity) obtained at two bleaching intensities gave an average apparent omega approximately 52 degrees. However, to adequately fit the F psi (0) data, inclusion of the theoretical effects of rapid (faster than the duration of the photobleaching pulse) fluorophore dynamics was also required. Best fits of the F psi (0) and F psi (infinity) data were obtained when the fluorophores were assumed to rapidly wobble within a cone of semiangle delta approximately 30 degrees-50 degrees while slowly rotating within an angular space defined by semiangle omega approximately 35 degrees-60 degrees. Subsequent analysis of the time- and psi-dependent changes in the post-bleach fluorescence curves F psi (t) gave apparent diffusion coefficients ranging from D approximately 10(-3) s-1 to 4 x 10(-2) s-1. PMID- 8427939 TI - Calculation and measurement of the dipole moment of small proteins: use of protein data base. AB - The dipole moments of small protein molecules were determined experimentally in order to validate the calculated dipole moments by previous investigators. We found that the agreements are satisfactory for some proteins. There are, however, many proteins for which the agreement is less than satisfactory. In order to find the cause of the disagreement, the dipole moments of these proteins were recalculated using the Brookhaven Protein Data Bank. We calculated the dipole moment due to fixed surface charges and the bond moments of all the carbonyl groups in main chain and side chains. The calculation consists of the mean moments and their mean square fluctuations. In addition, we investigated the effect of electrostatic interactions between charged sites for several proteins. These results show that incorporation of the interactions does not affect substantially the calculated dipole moments. The rms fluctuation of the dipole moment is found to be small but not negligible. In conclusion, recalculated dipole moments are in good agreement with the observed values. PMID- 8427940 TI - A Monte Carlo method for generating structures of short single-stranded DNA sequences. AB - A Monte Carlo method has been developed for generating the conformations of short single-stranded DNAs from arbitrary starting states. The chain conformers are constructed from energetically favorable arrangements of the constituent mononucleotides. Minimum energy states of individual dinucleotide monophosphate molecules are identified using a torsion angle minimizer. The glycosyl and acyclic backbone torsions of the dimers are allowed to vary, while the sugar rings are held fixed in one of the two preferred puckered forms. A total of 108 conformationally distinct states per dimer are considered in this first stage of minimization. The torsion angles within 5 kcal/mole of the global minimum in the resulting optimized states are then allowed to vary by +/- 10 degrees in an effort to estimate the breadth of the different local minima. The energies of a total of 2187 (3(7)) angle combinations are examined per local conformational minimum. Finally, the energies of all dinucleotide conformers are scaled so that the populations of differently puckered sugar rings in the theoretical sample match those found in nmr solution studies. This last step is necessitated by limitations in the theoretical methods to predict DNA sugar puckering accurately. The conformer populations of the individual acyclic torsion angles in the composite dimer ensembles are found to be in good agreement with the distributions of backbone conformations deduced from nmr coupling constants and the frequencies of glycosyl conformations in x-ray crystal structures, suggesting that the low energy states are reasonable. The low energy dimer forms (consisting of 150-325 conformational states per dimer step) are next used as variables in a Monte Carlo algorithm, which generates the conformations of single-stranded d(CXnG) chains, where X = A, T and n = 3, 4, 5. The oligonucleotides are built sequentially from the 5' end of the chain using random numbers to select the conformations of overlapping dimer units. The simulations are very fast, involving a total of 10(6) conformations per chain sequence. The potential errors in the buildup procedure are minimized by taking advantage of known rotational interdependences in the sugar-phosphate backbone. The distributions of oligonucleotide conformations are examined in terms of the magnitudes, positions, and orientations of the end-to-end vectors of the chains. The differences in overall flexibility and extension of the oligomers are discussed in terms of the conformations of the constituent dinucleotide steps, while the general methodology is discussed and compared with other nucleic acid model building techniques. PMID- 8427941 TI - Pharmacokinetics and pharmacodynamics of morphine-3-glucuronide in rats and its influence on the antinociceptive effect of morphine. AB - In this study the pharmacokinetics and pharmacodynamics of morphine-3-glucuronide (M3G) were investigated in rats after i.v. administration as a bolus dose (86.7 mumol kg-1) and as a constant rate infusion (2.9 mumol h-1) over 5 days. After the bolus dose, the clearance (Cl) was 12.1 +/- 0.6 ml min-1*kg, the volume of distribution at steady state (Vss) 1.68 +/- 0.89 l kg-1, the half-life of the first phase 13.2 +/- 1.8 min and the half-life of the second phase 11.6 +/- 7.7 h. After the constant rate infusion, Cl was 10.5 +/- 1.7 ml min-1*kg. The antagonistic effect of M3G on the antinociceptive effect of a bolus dose of morphine (35 mumol kg-1) was tested during steady state concentrations of M3G on day 4 and to M3G naive rats. No antinociceptive, hyperalgesic or withdrawal effects were observed as a result of M3G administration, but a significantly lower antinociceptive effect of morphine was found in the M3G infusion group compared to the control group. Systemically administered M3G antagonized the antinociceptive effect of morphine, but this cannot be the only explanation to the tolerance development observed after morphine administration. PMID- 8427942 TI - Pharmacokinetics and bioavailabilities of hymecromone in human volunteers. AB - Specific and ultrasensitive reverse-phase HPLC assays of the choleretic and biliary antispasmodic hymecromone (down to 0.05 ng ml-1) and its glucuronide, using fluorimetric detection, and sulfate metabolites using UV detection, were developed. Sodium salt solutions of 400 mg (over 3 min) and 800 mg (over 5 min) were infused i.v. into 6-8 normal human volunteers. The half-life of the major rate constant averaged 28 +/- 2 min (SE). Subsequently, less than 0.8 per cent of the dose was eliminated with terminal half-lives of 70-359 min. The apparent volume of distribution of hymecromone, referenced to the total plasma concentration, averaged 20.8 +/- 1.41 (Vc, central compartment volume) and 36.4 +/- 2.11 (Vss steady state volume). Hymecromone's total body clearance averaged 1413 +/- 89 ml min-1. The pharmacokinetics of hymecromone were dose-independent. Only 0.3 +/- 0.3 per cent unchanged hymecromone was renally excreted. Mostly dose independent glucuronidated drug (93 +/- 4 per cent of the dose) was excreted in the urine; a smaller amount was renally excreted as the sulfate (1.4 +/- 0.3 per cent of the dose). The oral bioavailability estimated from the relative areas under the hymecromone plasma concentration-time curves following oral and i.v. administration of hymecromone to six volunteer subjects showed no dose-dependence and was 1.8 +/- 0.6 per cent. However, an anomalous c. 200 per cent of the glucuronide produced by i.v. hymecromone was produced from orally administered hymecromone as determined from the ratio of the AUC values of glucuronide obtained after peroral and i.v. administration of the same dose of hymecromone to demonstrate a high first-pass effect and implicate renal glucuronidation. PMID- 8427943 TI - Study on pharmacokinetics of a new biliary excreted oral angiotensin converting enzyme inhibitor, temocapril (CS-622) in humans. AB - Despite the usefulness of angiotensin converting enzyme (ACE; EC 3.4.15.1) inhibitors for patients with renal insufficiency, some hesitation has been exercised in applying ACE inhibitors to the treatment of such patients because most ACE inhibitors are excreted mainly into the urine. In this context, development of an ACE inhibitor which is excreted into the bile has been sought. The pharmacokinetic properties of the novel ACE inhibitor, temocapril hydrochloride (temocapril HCl; CS-622), were investigated in six healthy volunteers. This drug is excreted mainly into the bile in animal studies. Temocapril HCl was given in a single dose of 0.5, 1.0, and 2.0 mg, and 36, 44, and 38 per cent of the administered drug was excreted in the feces and 17, 19, and 24 per cent in the urine as the de-esterified active diacid form (the diacid metabolite) within 48 h, respectively. The plasma ACE activity was markedly inhibited. No abnormal clinical findings suggestive of side-effects were observed. Thus, from the pharmacokinetic standpoint, temocapril HCl is expected to be a useful drug for patients with renal dysfunction. PMID- 8427944 TI - Pharmacokinetic analysis and anticonvulsant activity of two polyesteric prodrugs of valproic acid. AB - The pharmacokinetics of the following two polyesteric prodrugs of valproic acid (VPA) have been investigated: 1,4-butanediol divalproate (BDV) and glyceryl trivalproate (GTV). In addition, the anticonvulsant activity of these compounds has been evaluated and compared to that of VPA and valpromide (VPD). Valproic acid, and its two esteric derivatives were administered intravenously to six dogs at an equivalent dose (400 mg VPA) and their pharmacokinetics investigated. In the case of BDV, the biotransformation to VPA was complete, but in the case of GTV, it was only partial. Of the two investigated esteric prodrugs of VPA, only BDV demonstrated anticonvulsant activity and showed less neurotoxicity than VPA and VPD, and therefore had a better protective index. The anticonvulsant activity is explained on pharmacokinetic and pharmacodynamic grounds due to its complete conversion to VPA and the possible synergism in anticonvulsant activity between VPA and 1,4-butanediol. PMID- 8427945 TI - Effect of urine pH and ascorbic acid on the rate of conversion of methenamine to formaldehyde. AB - The kinetics of conversion of methenamine to the active form formaldehyde were studied in pooled urine samples at 37 degrees in the pH range 4.9-6.5. Using a method for the determination of both formaldehyde and unhydrolyzed methenamine, the rate of formaldehyde formation in urine was found to be apparent first order and was pH dependent. Bactericidal concentrations of formaldehyde (> 28 micrograms ml-1) were achieved in 3 h in urine of pH 6.0 containing methenamine at 750 micrograms ml-1. There was no difference in the in vitro rate of conversion of methenamine to formaldehyde between the urine collected from normal subjects and the urine from subjects administered ascorbic acid. The rates of degradation of the mandelate and hippurate salts in buffer systems of various pH values did not differ significantly from those of methenamine base in urine adjusted to the same pH. The half-life of methenamine conversion to formaldehyde increased approximately 20 times from 20 h at pH 5.0 to about 400 h at pH 6.5. The data suggest that unless the urine is maintained below pH 6 only a small fraction of methenamine would be converted daily to formaldehyde and, thus, may explain the need for large doses of this drug in patients. PMID- 8427946 TI - A method for calculating the mean absorption time of drugs undergoing reversible and first-pass metabolism. AB - A method has been derived for calculating the mean absorption time of an oral drug and its interconversion metabolite which is generated from the drug systemically and presystemically. The method evolves from the convolution integral and requires plasma AUC and AUMC values after separate intravenous administration of the drug and its interconversion metabolite and oral administration of the drug. It can also be used to calculate the mean input time of a drug undergoing reversible metabolism and administered by any other extravascular route. Results of a simulation study using both errorless and errant data indicate that, when the absorption rate constant of a drug or its interconversion metabolite is not much larger than the apparent elimination rate constant, the proposed method performs satisfactorily. However, when the absorption rate constant of a drug or its interconversion metabolite is much larger than the apparent elimination rate constant, the proposed method appears to be inaccurate. PMID- 8427947 TI - Pharmacokinetic interaction between valproic acid and phenobarbital. PMID- 8427948 TI - Blood to plasma ratio of mefloquine: interpretation and pharmacokinetic implications. AB - The blood to plasma ratio of the antimalarial mefloquine has been reported to be close to 1, while other reports indicate extensive accumulation in erythrocytes. This apparent contradiction has been resolved by a quantitative examination of the compensating effects of plasma protein binding of mefloquine which almost exactly matches the extent of mefloquine accumulation in erythrocytes. The observed blood to plasma ratio of about 1 arises as the result of a balance between extensive red cell uptake and extensive plasma protein binding. Some pharmacokinetic implications of the distribution of mefloquine within blood are outlined. PMID- 8427949 TI - [Intermittent self-catheterization in multiple sclerosis]. AB - Intermittent self-catheterization was taught to 70 patients with multiple sclerosis (MS) between 1.9.1989 and 31.1.1992 in Masku Neurological Rehabilitation Centre. All of them were interviewed in April 1992. Altogether 62 (89%) returned the questionnaires. The aims of the study were 1) to define the compliance of the patients to the treatment, and 2) to assess the effect of the catheterization on the quality of life of the patients. The conceptual framework used for the study was self care concept. Bladder symptoms had been present for seven years (mean) and the average time after the diagnosis of MS was ten years. Twenty eight (28) patients discontinued the treatment after having catheterized for 11 months in average. The main reason (50%) for discontinuation was normalization of the bladder function. The patients reported reduction in urgency symptoms, incontinence of urine and difficulties in emptying the bladder. All these changes were statistically significant. The number of urinary infections was slightly increased (not reaching statistical significance). However, the infections caused less subjective discomfort during the treatment than before it. The urinary symptoms interfered with the personal activities of the patients in a variety of ways. As many as 79% of the respondents reported that their quality of life was improved by the intermittent self-catheterization. The treatment allowed the patients to resume their personal, social -- and in some cases occupational- activities of daily living. Significant improvement was reported in f.e. family life, marital relationships, sexual life and the quality of night sleep. PMID- 8427950 TI - [Public health care's changing environment: 1990 statistics]. PMID- 8427951 TI - [The future of public health care hospitals]. AB - The subvention from the government for public health care system hospitals will decrease in Finland 1993. This causes that in future hospitals will work more like private enterprises. Expenses have to be covered by incomes from sales of health care services. Hospitals have to be able to estimate the demand of their services and the situation caused by the competition. They have to be able to create new profile and also be able to market their services. The presupposition for all things mentioned above is the creativity of people working in different wards. Creativity helps to obtain the same or even better quality of services with smaller resources. PMID- 8427952 TI - [Development of ICN's Code of Ethics]. AB - Development of the ICN Code for Nurses (Ethical Concepts Applied to Nursing) was explored from the Finnish perspective. The article was mainly based on the archival sources of the Finnish Federation of Nurses and on the ICN publications. The 1953, 1965 and 1973 Codes were briefly discussed. It was observed that the Federation never explicitly integrated the Code for Nurses into its decision making. PMID- 8427953 TI - Coagulation assembly on leukocytes in transmembrane signaling and cell adhesion. PMID- 8427954 TI - Activation of factor XI in plasma is dependent on factor XII. AB - The activation of factor XI initiates the intrinsic coagulation pathway. Until recently it was believed that the main activator of factor XI is factor XIIa in conjunction with the cofactor high molecular weight kininogen on a negatively charged surface. Two recent reports have presented evidence that in a purified system factor XI is activatable by thrombin together with the soluble polyanion dextran sulfate. To assess the physiological relevance of these findings we studied the activation of factor XI in normal and factor XII-deficient plasma. We used either kaolin/cephalin or dextran sulfate as a surface for the intrinsic coagulation pathway, tissue factor to generate thrombin via the extrinsic pathway, or the addition of alpha-thrombin directly. 125I-factor XI, added to factor XI-deficient plasma at physiologic concentrations (35 nmol/L), is rapidly cleaved on incubation with kaolin. The kinetics appear to be exponential with half the maximum cleavage at 5 minutes. Similar kinetics of factor XI cleavage are seen when 40 nmol/L factor XIIa (equal to 10% of factor XII activation) is added to factor XII-deficient plasma if an activating surface is provided. Tissue factor (1:500) added to plasma did not induce cleavage of factor XI during a 90 minute incubation, although fibrin formation within 30 seconds indicated that thrombin was generated via the extrinsic pathway. Adding 1 mumol/L alpha-thrombin (equivalent to 50% prothrombin activation) directly to factor XII deficient or normal plasma (with or without kaolin/cephalin/Ca2+ or dextran sulfate) led to instantaneous fibrinogen cleavage, but again no cleavage of factor XI was observable. We conclude that in plasma surroundings factor XI is not activated by thrombin, and that proposals of thrombin initiation of the intrinsic coagulation cascade are not supportable. PMID- 8427955 TI - A low-affinity human granulocyte-macrophage colony-stimulating factor/murine erythropoietin hybrid receptor functions in murine cell lines. AB - To identify domains in hematopoietic growth factor receptors that are important for signal transduction, a hybrid receptor (GMER) was constructed by splicing the DNA of the entire extracellular and transmembrane domains of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor alpha 2 subunit (GMR) to the cytoplasmic domain of the murine erythropoietin receptor (mEpoR). The hybrid receptor was introduced into the interleukin-3 factor dependent murine hematopoietic cell line Ba/F3. Cells that expressed high receptor numbers were selected by cell sorting using phycoerythrin-labeled human GM-CSF. Immunoprecipitation of GMER from Ba/F3 cells showed a band with an Mr of 105,000 daltons. Human GM-CSF binding to Ba/F3 cells that expressed GMER showed a kd of 3.0 nmol/L and 475 binding sites/cell, while the same cells that expressed GMR had 300 sites/cell and a kd of 3.5 nmol/L. The proliferative response to GM CSF of Ba/F3 cells that expressed GMER showed 1/2 maximal cell growth (as measured by 3H-thymidine incorporation) at a GM-CSF concentration of 2.5 x 10(-8) mol/L. When cultured in human GM-CSF, Ba/F3-GMER cells expressed cell surface glycophorin. Similar results were obtained with Ba/F3 cells transfected with the mEpoR and cultured in erythropoietin. Expression of GMR plus the human GM-CSF receptor beta chain in the same cell line also resulted in human GM-CSF stimulated proliferation; however, cell surface glycophorin was not detected. These data show that a low-affinity GM-CSF/Epo hybrid receptor can promote GM-CSF dependent proliferation and can induce the expression of glycophorin, an erythroid-specific protein. PMID- 8427956 TI - Translocation breakpoints are clustered on both chromosome 8 and chromosome 21 in the t(8;21) of acute myeloid leukemia. AB - The t(8;21)(q22;q22) is consistently associated with acute myeloid leukemia (AML) M2. Recent data have suggested that breakpoints on chromosome 21 are clustered within a single intron of a novel gene, AML1, just downstream of a region of homology to the runt gene of D melanogaster. In this report, we confirm rearrangement at the same location in at least 12 of 18 patients with t(8;21). Furthermore, we have isolated recombinant clones spanning the breakpoint regions on both the der(8) and the der(21) from one patient. By using a chromosome 8 probe derived from these clones, we show that t(8;21) breakpoints are also clustered on chromosome 8. PMID- 8427957 TI - Treatment of late bone marrow relapse in children with acute lymphoblastic leukemia: a Pediatric Oncology Group study. AB - Children with acute lymphoblastic leukemia (ALL) who have completed 2.5 to 3 years of initial chemotherapy have an off-therapy relapse rate of approximately 20%. In an attempt to improve the survival of children with a late bone marrow (BM) relapse (ie, occurring greater than 6 months after cessation of primary therapy), the Pediatric Oncology Group designed a randomized study to compare the efficacy of doxorubicin/prednisone and cytarabine/teniposide in a multidrug retreatment chemotherapy program. Treatment consisted of remission reinduction with vincristine, prednisone, and doxorubicin, central nervous system prophylaxis with triple intrathecal chemotherapy, and continuation therapy (for 132 weeks) with alternating cycles of oral 6-mercaptopurine/methotrexate and intravenous vincristine/cyclophosphamide. Patients received intermittent courses of either prednisone/doxorubicin (regimen 1) or teniposide/cytarabine (regimen 2) during continuation therapy and a late intensification phase with either vincristine, prednisone, and doxorubicin (regimen 1) or teniposide and cytarabine (regimen 2). One hundred two of 105 evaluable patients (97%) achieved a second complete remission. Twenty-eight of 50 patients on regimen 1 have failed compared with 28 or 52 patients on regimen 2 (log-rank analysis, P = .68), indicating that this trial was inconclusive as to which treatment regimen was superior. The overall 4 year event-free survival for children with a late BM relapse was 37% +/- 6%. Age less than 10 years at initial diagnosis (P < or = .001), white blood cell count less than 5,000/microL at relapse (P = .036) and duration of first remission greater than 54 months (P = .039) were independently associated with a more favorable outcome. While the randomized trial was inconclusive, prolonged second complete remissions were secured in more than one-third of children with a late BM relapse of ALL. The prognostic factors identified may help select children with a late BM relapse who can be successfully retreated with chemotherapy alone. PMID- 8427958 TI - Basic fibroblast growth factor expression in human bone marrow and peripheral blood cells. AB - We have shown previously that basic fibroblast growth factor (bFGF) is a mitogen for human bone marrow (BM) stromal cells and that bFGF stimulates myelopoiesis in primary BM cultures. In this article, we demonstrate the presence of bFGF in two cell lineages in human BM and peripheral blood as well as the deposition of bFGF into the extracellular matrix of BM stromal cell cultures. In immunofluorescence experiments on BM and peripheral blood smears, megakaryocytes and platelets stained strongly for bFGF, whereas weaker staining was observed in immature and mature cells of the granulocyte series. The presence of bFGF in platelets was confirmed by enzyme-linked immunosorbent assay as well as by immunoprecipitation followed by immunoblotting. bFGF was synthesized by BM stromal cell cultures and was found either cell associated or localized in the nucleus and the nucleoli, and its location was dependent on the fixation procedure used. Addition of exogenous bFGF to stromal cells showed the presence of extracellular binding molecules for this cytokine. bFGF could be released from these sites by soluble heparin or phosphatidylinositol-specific phospholipase C. This study supports the role of bFGF as a stromal cell mitogen and stimulator of myelopoiesis. The data indicate that the stromal cells produce bFGF and that their extracellular matrix can serve as a reservoir for this growth factor. In addition, the results suggest a possible involvement of bFGF in platelet function as well as in megakaryocytopoiesis. PMID- 8427959 TI - A role for manganese superoxide dismutase in radioprotection of hematopoietic stem cells by interleukin-1. AB - Pretreatment with interleukin-1 (IL-1) has been shown to protect mice from the myelotoxicity associated with irradiation via a mechanism potentially mediated through the induction of the antioxidant enzyme manganese superoxide dismutase (MnSOD). In this study, we have compared the ability of IL-1 to induce MnSOD mRNA in murine bone marrow cells and human cell lines with its ability to protect these cells against the damaging effects of ionizing radiation. Bone marrow cells obtained from mice 6 hours after a single injection of IL-1 demonstrate a dose dependent increase in the expression of MnSOD RNA. In this same study, IL-1 was also shown to be radioprotective when given to mice 20 hours before lethal irradiation. Similarly, in vitro treatment with IL-1 of bone marrow cells isolated from 5-fluorouracil-treated mice results in elevated levels of MnSOD RNA. Pretreatment with IL-1 also protected bone marrow long-term culture initiating cells capable of reconstituting irradiated stromal cultures from an irradiation insult. Furthermore, IL-1-treated human bone marrow cells display both elevated MnSOD RNA and protein levels when compared with media controls. The human A375 melanoma, A549 adenocarcinoma, and factor-dependent TF-1 leukemic cell lines demonstrate low basal MnSOD RNA levels that increase following treatment with IL-1. For the A375 cells, this correlates with increased MnSOD protein expression and radioprotection by IL-1 using a colony assay. In contrast, the chronic myelogenous leukemic cell line, K562, displays a high basal MnSOD RNA level, and this RNA expression is not further increased by IL-1 treatment. In addition, these cells are comparatively radioresistant and are not further protected by IL-1 treatment. Finally, the Mo-7 cell line displays a low basal level of MnSOD RNA that correlates with a high sensitivity to irradiation and IL 1 pretreatment has no effect on MnSOD RNA levels. Our results indicate that increased radioprotection by IL-1 correlates with the induction of the antioxidant enzyme MnSOD and this induction may be an important factor in IL-1 radioprotection. PMID- 8427960 TI - Homocysteine inhibits von Willebrand factor processing and secretion by preventing transport from the endoplasmic reticulum. AB - Intracellular protein transport in endothelial cells is selectively inhibited by homocysteine, a thiol amino acid associated with both thrombosis and atherosclerosis. In a previous study, homocysteine decreased cell surface expression of the surface transmembrane glycoprotein thrombomodulin without decreasing secretion of another endothelial cell protein, plasminogen activator inhibitor-1. To define further the effects of homocysteine on protein transport, we examined the processing and secretion of the multimeric glycoprotein von Willebrand factor (vWF) in human umbilical vein endothelial cells. Incubation with 2 mmol/L homocysteine resulted in complete loss of vWF multimers and prevented asparagine-linked oligosaccharide maturation, propeptide cleavage, and secretion; these effects are consistent with impaired exit from the endoplasmic reticulum (ER). Dimerization was only partially inhibited, suggesting that homocysteine causes retention of provWF in the ER without preventing dimer formation. In pulse-chase incubations, intracellular provWF was degraded before exiting the ER in homocysteine-treated cells. Homocysteine also inhibited the processing and secretion of a carboxyl-terminal truncation mutant of human provWF expressed in rat insulinoma cells, indicating that retention in the endoplasmic reticulum can be mediated by regions of provWF apart from the carboxyl-terminal 20-Kd segment. These results suggest that retention of secretory proteins in the ER is regulated by redox mechanisms and imply that the intracellular transport of multiple endothelial cell proteins may be altered in patients with homocystinuria. PMID- 8427961 TI - Increased thrombin generation in a child with a combined factor IX and protein C deficiency. AB - We report a quantitative protein C deficiency combined with a factor IX deficiency in a one-year-old boy. The inheritance of the two deficiency states was independent, the factor IX defect coming from the mother and the protein C defect from the father. Both factor IX activity and antigen were below 1%, and protein C activity as well as antigen were close to 27% of normal values. This association raises a real therapeutic and prognostic question. Protein C deficiency is indeed associated with a significant thrombotic risk and some factor IX concentrates seem to carry a potential thrombogenicity, particularly following infusion of repeated doses. We evaluated in this patient the potential activation of the coagulation system by measuring the levels of prothrombin fragment F1 + 2 at the basal state and after a single administration of 20 U/kg of a high purity factor IX concentrate. We found an unexpected basal activation of the hemostatic system before infusion (F1 + 2 = 1.6 nmol/L), which further increased during 8 hours. Despite the clinical predominant expression of the hemophilic trait, our results seem to assess the biologic prevalence of the protein C deficiency. This emphasizes the need for a careful follow-up after infusions of repeated doses of factor IX, as used during a surgical procedure. Furthermore, this raises the question of the prognosis because the risk of thrombotic manifestations associated with a protein C deficiency increases with age. Finally, these results highlight a part of the in vivo activation process of prothrombin in case of failure of the intrinsic pathway of coagulation. The protein C defect seems to be responsible for an upregulation of the prothrombin activation through the extrinsic pathway. PMID- 8427962 TI - Pharmacokinetic and thrombolytic properties of chimeric plasminogen activators consisting of a single-chain Fv fragment of a fibrin-specific antibody fused to single-chain urokinase. AB - The pharmacokinetic and thrombolytic properties were determined of two recombinant single-chain chimeric plasminogen activators (PA) consisting of u-PA 33k, a low-molecular weight derivative of single-chain urokinase-type PA (scu-PA) comprising amino acids Ala132 through Leu411, and of either a single-chain variable region fragment (Fv) derived from the fibrin fragment D-dimer-specific monoclonal antibody MA-15C5 (K12G0S32) or of the deglycosylated single-chain Fv fragment obtained by substitution of Asn88 with Glu (K12G2S32). Following bolus injection in hamsters, clearances of recombinant scu-PA (rscu-PA) and of K12G0S32 were similar. In contrast, clearance of K12G2S32 was fourfold slower than that of rscu-PA. The thrombolytic potency (percent lysis per u-PA administered in milligrams per kilogram body weight) and specific thrombolytic activity (percent lysis per microgram per milliliter steady-state plasma u-PA antigen level) of these compounds were studied in hamsters with an experimental pulmonary embolus consisting of a human plasma clot injected via the jugular vein. The doses of K12G0S32 and K12G2S32 required to obtain maximal rate of clot lysis were sixfold and 11-fold lower than that of rscu-PA. The steady-state u-PA-related plasma antigen levels of K12G0S32 and K12G2S32 required to obtain maximal rate of clot lysis were 10-fold and fourfold lower than that of rscu-PA. Thus, targeting of K12G0S32 to the clot surface by means of its glycosylated Fv fragment results in a 10-fold increase of its specific thrombolytic activity and sixfold increase of its thrombolytic potency as compared with those of rscu-PA. Targeting of K12G2S32 to the clot surface by means of its deglycosylated Fv fragment results in only a twofold increase of its thrombolytic activity. However, its fourfold slower clearance, combined with its twofold higher specific thrombolytic activity, results in an 11-fold increase of its thrombolytic potency over that of rscu-PA. These findings indicate that the thrombolytic potency of chimeric antibody targeted PA may be increased by increasing the specific thrombolytic activity, reducing the clearance, or both. PMID- 8427963 TI - Phosphorylation of factor Va and factor VIIIa by activated platelets. AB - Platelet activation leads to the incorporation of 32[PO4(2-)] into bovine coagulation factor Va and recombinant human factor VIII. In the presence of the soluble fraction from thrombin-activated platelets and (gamma-32P) adenosine triphosphate, radioactivity is incorporated exclusively into the M(r) = 94,000 heavy chain (H94) of factor Va and into the M(r) = 210,000 to 90,000 heavy chains as well into the M(r) = 80,000 light chain of factor VIII. Proteolysis of the purified phosphorylated M(r) = 94,000 factor Va heavy chain by activated protein C (APC) gave products of M(r) = 70,000, 24,000, and 20,000. Only the intermediate M(r) = 24,000 fragment contained radioactivity. Because the difference between the M(r) = 24,000 and M(r) = 20,000 fragments is located on the COOH-terminal end of the bovine heavy chain, phosphorylation of H94 must occur within the M(r) = 4,000 peptide derived from the carboxyl-terminal end of H94 (residues 663 through 713). Exposure of the radioactive factor VIII molecule to thrombin ultimately resulted in a nonradioactive light chain and an M(r) = 24,000 radioactive fragment that corresponds to the carboxyl-terminal segment of the A1 domain of factor VIII. Based on the known sequence of human factor VIII, phosphorylation of factor VIII by the platelet kinase probably occurs within the acidic regions 337 through 372 and 1649 through 1689 of the procofactor. These acidic regions are highly homologous to sequences known to be phosphorylated by casein kinase II. Results obtained using purified casein kinase II gave a maximum observed stoichiometry of 0.6 mol of 32[PO4(2-)]/mol of factor Va heavy chain and 0.35 mol of 32[PO4(2-)]/mol of factor VIII. Phosphoamino acid analysis of phosphorylated factor Va by casein kinase II or by the platelet kinase showed only the presence of phosphoserine while phosphoamino acid analysis of phosphorylated factor VIII by casein kinase II showed the presence of phosphothreonine as well as small amounts of phosphoserine. The platelet kinase responsible for the phosphorylation of the two cofactors was found to be inhibited by several synthetic protein kinase inhibitors. Finally, partially phosphorylated factor Va was found to be more sensitive to APC inactivation than its native counterpart. Our findings suggest that phosphorylation of factors Va and VIIIa by a platelet casein kinase II-like kinase may downregulate the activity of the two cofactors. PMID- 8427964 TI - Degradation of von Willebrand factor in patients with acquired clinical conditions in which there is heightened proteolysis. AB - The behavior of plasma von Willebrand factor (vWF) in patients with acute leukemia (n = 5), decompensated cirrhosis (n = 10), and acute pancreatitis (n = 5) was investigated to evaluate whether the systemic proteolytic states associated with these diseases had affected the structure and function of the molecule. vWF antigen and, to a lesser degree, ristocetin cofactor activity in patient plasma were high. Multimeric analysis of plasma vWF revealed loss of high molecular weight multimers. The subunit composition and proteolytic pattern of vWF immunopurified from patient plasmas and reduced were studied by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis followed by transblotting and probing with monoclonal antibodies that distinguish cleavages caused by plasmin from those caused by other proteases. There was marked reduction of the relative concentration of the native vWF subunit of 225 Kd in all patient groups, indicating heightened cleavage of the protein. The concentrations of 189- and 140-Kd vWF fragments, normally present in plasma, were increased in cirrhosis and pancreatitis but not in leukemia. Novel fragments, ranging in size from less than 225 to approximately 120 Kd were present in leukemia and cirrhosis, including plasmin-generated fragments of 176 and 145 Kd. These data indicate that in clinical conditions in which there is heightened proteolysis vWF is degraded in vivo by plasmin and other proteases. Degraded vWF may be less effective than native vWF in supporting primary hemostasis, thereby being a cofactor in the multifactorial bleeding diathesis accompanying systemic proteolytic states. PMID- 8427965 TI - Quantitation of activated factor VII levels in plasma using a tissue factor mutant selectively deficient in promoting factor VII activation. AB - Although the majority of factor VII (FVII) circulates in the zymogen form, low levels of activated factor VII (FVIIa) have been postulated to exist in plasma and to serve a priming function for triggering of the clotting cascade. However, direct measurement of plasma FVIIa has not previously been possible. We have quantified plasma FVIIa levels using a novel, highly sensitive assay that is free from interference by FVII. Specificity of this clot-based assay results from the use of a mutant tissue factor that is selectively deficient in promoting FVII activation, but retains FVIIa cofactor function. In normal adults, FVIIa was found to be present in plasma (mean: 3.6 ng/mL) with considerable variation between individuals (range: 0.5 to 8.4 ng/mL). FVIIa levels were only loosely correlated with FVII coagulant activity, but were elevated in pregnancy and reduced with oral anticoagulant therapy. Incubation of plasma on ice in glass containers (cold activation) resulted in substantial FVIIa generation. Measurement of plasma forms of factor VII is of potential clinical importance because elevated FVII coagulant activity has been implicated as a significant risk predictor for ischemic heart disease. Clinically, this new assay will now permit direct assessment of the role of plasma FVIIa in thrombotic disorders. PMID- 8427966 TI - Immunoglobulins from normal sera bind platelet vinculin and talin and their proteolytic fragments. AB - Our previous finding that normal serum immunoglobulins bind to internal platelet proteins on Western blots led us to further identify these proteins and determine the possible significance of autoantibodies against them. A 95-Kd protein reactive with immunoglobulins in most normal sera and easily confused with gpIIIa was shown to be a fragment of vinculin generated by calpain proteolysis. Identity was established by peptide sequencing of the protein purified from platelets stored without specific protease inhibitors. Normal immunoglobulins bound intact vinculin (117 Kd) and metavinculin (152 Kd), and their 105-, 95-, and 80- to 85 Kd proteolytic fragments. IgG in 89%, and IgA and IgM in 100% of normal sera reacted in titers of 10 to 1,000 with purified vinculin. In addition, IgG in 79%, and IgA and IgM in 93% of normal sera reacted in titers of 10 to 5,000 with talin (235 Kd), another cytoskeletal protein, and its 200-Kd proteolytic fragment. IgGs in sera from animals of several different phylogenetic classes also reacted with human vinculin and talin on Western blots. Frequency of occurrence, titers, and classes of antivinculin and antitalin autoantibodies in patients with thrombocytopenia did not differ discernibly from those of normal individuals. These antibodies had no effect on platelet aggregation or clot retraction, and no apparent pathogenic significance, but their widespread presence and the variability in extent of proteolysis of platelet preparations used for Western blots can complicate interpretation of patterns obtained with sera from patients with presumed immune-mediated thrombocytopenias. PMID- 8427967 TI - Interferon-alpha alters spectrin organization in normal and leukemic human B lymphocytes. AB - Interferon-alpha (IFN-alpha) regulates the growth, differentiation, and recirculation of normal and malignant B lymphocytes. In this report we examine the effects of IFN-alpha on the distribution of the cytoskeletal protein spectrin in peripheral blood B lymphocytes from normal donors and patients diagnosed with chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL). Exposure of normal and leukemic B cells to IFN-alpha in vitro was shown by immunofluorescence microscopy to cause a dose-dependent increase in the percentage of cells containing discrete focal accumulations of spectrin, ie, a single large aggregate or cap-like structure near the plasma membrane. Although the magnitude of this effect was variable among individual patient samples, in some experiments IFN alpha induced a fourfold increase in the percentage of leukemic B cells exhibiting focal accumulations of spectrin. Spectrin reorganization induced by IFN-alpha was abrogated by the protein synthesis inhibitor cycloheximide. In addition, IFN-alpha increased the total cellular content of spectrin in B-CLL cells by approximately twofold to fourfold. Finally, a role for protein kinase C in mediating the effects of IFN-alpha on spectrin's organization is implicated by studies in which calphostin C inhibited the IFN-induced focal accumulation of spectrin. Taken together, these studies suggest that the immunomodulatory activities of IFN-alpha in normal and malignant B cells involve a change in the organization of the spectrin-based cytoskeleton. PMID- 8427968 TI - Expression of syndecan regulates human myeloma plasma cell adhesion to type I collagen. AB - The syndecans comprise a family of integral membrane proteoglycans that regulate cell behaviors by binding to extracellular matrix and binding growth factors. In mouse blood cells, syndecan expression is restricted to cells of the B-cell lineage where it is expressed by pre-B cells and plasma cells, but is absent from circulating B cells. In the present study, we examined the expression, structure, and function of syndecan on human myeloma cell lines and myeloma patient bone marrow cells. On myeloma cells, syndecan is a small (modal relative molecular mass [M(r)] = 120 Kd) heparan sulfate proteoglycan localized at the cell surface. Syndecan was detected by immunodot blotting on 7 of 10 human myeloma cell lines and by reverse transcriptase polymerase chain reaction on 10 of 14 patient samples. Cell binding assays show that myeloma cells expressing syndecan bind to type I collagen via heparan sulfate chains, while those cell lines not expressing syndecan do not bind to collagen. Furthermore, the cell lines expressing syndecan were negative for CD19 and CD45 staining, indicating that syndecan expression is restricted to tumors having a well-differentiated phenotype. We conclude that syndecan acts as a matrix receptor on human myeloma cells but is not expressed by all tumors, suggesting that syndecan may participate in regulating myeloma cell adhesion to the bone marrow stromal matrix. PMID- 8427969 TI - Immunoglobulin heavy chain gene VH-D junctional diversity at diagnosis in patients with acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) represents the clonal outgrowth of transformed hematopoietic progenitor cells. We have found that blast cells in some cases of B precursor cell ALL contain Ig heavy chain gene rearrangements with considerable diversity at the junctions of the variable (VH), diversity (D), and joining (JH) regions. This diversity consists of heterogeneous nucleotide sequences at the VH D and, less frequently, the D-JH junctions. In two cases, different VH segments were attached to the same D-JH rearrangement. In all cases studied there was a much higher than expected frequency of nucleotide sequence changes in the VH segment. At least three mechanisms may produce these changes in different cases: (1) continuing rearrangement of the heavy chain gene, in some cases by VH addition to a preexisting D-JH; (2) VH replacement; and (3) an open-and-shut mechanism. These findings suggest that an active VDJ recombinase system is present at the time of transformation in a high percentage of ALLs. An active recombinase in the rapidly growing leukemic cell population could lead to genomic instability. PMID- 8427970 TI - Dye-mediated photolysis is capable of eliminating drug-resistant (MDR+) tumor cells. AB - We evaluated the potential role of photoradiation therapy with a benzoporphyrin derivative, monoacid ring A (BPD-MA), and dihematoporphyrin ether (DHE), for the ex vivo purging of residual tumor cells from autologous bone marrow (BM) grafts. BPD-MA and DHE photosensitizing activity was tested against two human large-cell lymphoma cell lines and colony-forming unit-leukemia (CFU-L) derived from patients with acute myelogenous leukemia (AML). In mixing experiments, 4-log elimination of tumor cell lines was observed after 1 hour of incubation with 75 ng/mL of BPD-MA or 30 minutes of treatment with 12.5 micrograms/mL of DHE followed by white light exposure. By comparison, using the same concentration of BPD-MA, the mean recovery of normal BM progenitors was 4% +/- 0.8% (mean +/- SD) for granulocyte-macrophage colony-forming unit (CFU-GM) and 5% +/- 0.8% for burst forming unit-erythroid (BFU-E). Similarly, DHE treatment resulted in the recovery of 5.2% +/- 2% and 9.8% +/- 3% of CFU-GM and BFU-E, respectively. Furthermore, equivalently cytotoxic concentrations of both DHE and BPD-MA and light were found not to kill normal pluripotent stem cells in BM, as demonstrated by their survival in two-step long-term marrow culture at levels equal to untreated controls. The T-lymphoblastic leukemia cell line CEM and its vinblastine (VBL) resistant subline CEM/VBL, along with the acute promyelocyte leukemia cell line HL-60 and its vincristine (VCR)-resistant subline HL-60/VCR, were also tested. BPD-MA at 75 ng/mL was able to provide a greater than 4-log elimination of the drug-sensitive cell lines, but only a 34% and 55% decrease of the drug-resistant HL-60/VCR and CEM/VBL cell lines, respectively. On the contrary, 12.5 micrograms/mL of DHE reduced the clonogenic growth of all the cell lines by more than 4 logs. Further experiments demonstrated decreased uptake of both BPD-MA and DHE by the resistant cell lines. However, all the cell lines took up more DHE than BPD-MA under similar experimental conditions. Our results demonstrate the preferential cytotoxicity of BPD-MA and DHE toward neoplastic cell lines and CFU L from AML patients. In addition, DHE was slightly more effective in purging tumor cells expressing the p-170 glycoprotein. These results suggest that photoradiation with DHE would be useful for in vitro purging of residual drug resistant leukemia and lymphoma cells. PMID- 8427971 TI - Analysis of mutant NADH-cytochrome b5 reductase: apparent "type III" methemoglobinemia can be explained as type I with an unstable reductase. AB - A patient in Kurobe, Japan, was previously reported to have a new class of hereditary methemoglobinemia, type III. In this patient, NADH cytochrome b5 reductase deficiency was observed in lymphocytes and platelets as well as in erythrocytes, but this was not associated with mental retardation. A base change was identified in the gene for NADH cytochrome b5 reductase, causing an amino acid substitution from Leu-148 to Pro. In the present study, the mutant enzyme (Leu-148-->Pro) was expressed in Escherichia coli, purified, and characterized. The mutant enzyme retained about 60% of the catalytic activity of the wild type, but was remarkably heat unstable. By incubating the mutant enzyme at 42 degrees C for 10 minutes, 80% of the enzyme activity was lost, whereas the wild-type enzyme lost < 20% activity after incubation at 50 degrees C for 30 minutes. Another mutant in which Leu-148 was replaced by Ala was prepared to establish the role of the residue. This mutant was apparently less heat stable than the wild type, implying a structural role for Leu-148. Reinvestigation of the enzyme activity in the blood cells and fibroblasts of the type III Kurobe patient, revealed that about 40% of the normal activity was detected in these cells, in contrast to the previous report. Thus, this patient reported previously as having hereditary meth hemoglobinemia type III was shown to have type I. PMID- 8427972 TI - Human erythrocyte acetylcholinesterase bears the Yta blood group antigen and is reduced or absent in the Yt(a-b-) phenotype. AB - The Cartwright (Yt) blood group antigens have previously been shown likely to reside on a phosphatidylinositol-linked erythrocyte membrane protein. In this study, an unusual individual whose red blood cells (RBCs) were of the previously unreported Yt(a-b-) phenotype were used, along with normal Yt(a+) cells, to investigate serologically and biochemically the relationship of the Yta antigen to known phosphatidylinositol-linked erythrocyte proteins. Yt(a-b-) RBCs expressed normal amounts of various phosphatidyl-inositol-linked proteins except acetylcholinesterase. Further, human anti-Yta reacted with acetylcholinesterase in immunoprecipitation and immunoblotting studies. Thus, acetylcholinesterase is now identified as the protein bearing the Yt blood group antigens. PMID- 8427973 TI - Effect of uremia on the membrane transport characteristics of red blood cells. AB - Even though there is extensive evidence that uremia affects the fragility and deformability of red blood cells (RBCs), essentially all data on the RBC membrane permeability have been obtained with nonuremic blood. Permeability data were obtained for creatinine and uric acid, two metabolites of interest in hemodialysis, using a stirred ultrafiltration device with direct cell- and protein-free sampling. Experiments examined the effects of temperature and suspending phase on solute transport for both normal and uremic blood cells. Creatinine and uric acid transport from normal RBCs at 37 degrees C were characterized by saturation half-times of 40 +/- 10 minutes and 54 +/- 12 minutes, respectively. The corresponding half-times for uremic cells were significantly longer, 94 +/- 26 minutes and 180 +/- 38 minutes. Data indicated that the slower rate of creatinine transport in uremic blood was caused by an alteration in the RBC membrane, while the reduction in uric acid transport was associated with alterations in the uremic plasma. The temperature dependence of the RBC permeability was also much less pronounced for uremic cells for both solutes. These results provide important insights into the effects of uremia on the RBC membrane permeability, and have important implications for dialysis. PMID- 8427974 TI - Human immunodeficiency virus-related conditions in children and adults with hemophilia: rates, relationship to CD4 counts, and predictive value. AB - To further elucidate the natural history of human immunodeficiency virus (HIV) infection, we studied intermediate HIV-related conditions occurring before acquired immunodeficiency syndrome (AIDS) in a prospectively observed multicenter cohort of 738 HIV-infected persons with hemophilia. We analyzed the frequency in adults and children of common HIV-related conditions and the relative risk of AIDS after occurrence of these conditions, controlling for age at seroconversion and the percentage of CD4+ lymphocytes. Thrombocytopenia was the most frequently observed condition with cumulative incidences of 43% +/- 7% in adults and 27% +/- 6% in children and adolescents by 10 years after seroconversion. Oral candidiasis, fever, weight loss, and non-AIDS pneumonia were two to four times more common in adults than children, whereas herpes zoster risk was similar in the two age groups. HIV-related conditions were infrequent during the first 4 years of infection, particularly in children. With the exception of thrombocytopenia, mean CD4 counts were less than 350 cells/microL at the onset of the conditions. The relative hazard of AIDS after oral candidiasis was 18 in children and 3.8 in adults. Relative hazard in adults was also increased after persistent fever (10), weight loss (3.2), and non-AIDS pneumonia (2.2). Herpes zoster and thrombocytopenia were not significantly associated with AIDS in either age group. We conclude that intermediate HIV-related conditions occur more frequently in adults than in children with hemophilia. Persistent fever is the strongest predictor of AIDS in adults, whereas oral candidiasis is the strongest predictor in children. These findings should facilitate the design and conduct of clinical trials as well as the management of HIV-infected children and adults. PMID- 8427975 TI - Determination of human platelet antigen frequencies in the Dutch population by immunophenotyping and DNA (allele-specific restriction enzyme) analysis. AB - Platelets from 200 random Dutch blood donors were typed for the human platelet alloantigens HPA-1 to -5 recognized at present and for Naka. Naka is an epitope on glycoprotein IV, not expressed on the platelet of individuals with hereditary GP IV deficiency. Platelet immunofluorescence and monoclonal antibody-specific immobilization of platelet antigens (MAIPA) were applied for this purpose. The observed phenotype frequencies were 97.86% and 28.64% for HPA-1a and -1b, 100% and 13.15% for HPA-2a and -2b, 80.95% and 69.84% for HPA-3a and -3b, 100% and 0% for HPA-4a and -4b, 100% and 19.7% for HPA-5a and HPA-5b, respectively. Platelets from all donors reacted with the anti-Naka antibodies. To determine the gene frequencies for the HPA-1, HPA-2 and HPA-3 systems directly, DNA from 98 of these donors was isolated from peripheral blood mononuclear leucocytes and specific fragments were amplified by polymerase chain reaction (PCR). The fragments were analyzed using allele-specific restriction enzymes (ASRA). In all amplified PCR products an "internal control" for each assay, ie, a restriction site for the applied enzyme independent from the phenotype of the donor was present. In all donors tested, phenotypes, as determined by serological methods and genotypes, directly determined by the ASRA, were identical. Thus, the PCR-ASRA described in this report is a practical and reliable technique for the determination of alleles that code for platelet antigen allotypes, at least in the Dutch population. PMID- 8427976 TI - Quantitative assessment of posttransplant host-specific interleukin-2-secreting T helper cell precursors in patients with and without acute graft-versus-host disease after allogeneic HLA-identical sibling bone marrow transplantation. AB - Recent studies in mice and humans have emphasized an important contribution of host-reactive minor histocompatibility antigen (mH)-specific lymphokine-secreting donor T-helper cells (Th) for the induction of acute graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation (BMT). By using limiting dilution (LD) and clonal specificity analyses, we investigated in 14 patients with and without acute GVHD after non-T-depleted HLA-identical sibling BMT whether posttransplant host-reactive mH-specific interleukin-2 (IL-2)-secreting Th are involved in the development of clinically significant acute GVHD and the establishment of tolerance. At different time intervals posttransplant (I, days 0 through 45; II, days 45 through 90; III, days 90 through 180), host-specific IL-2 secreting Th-precursors (Th-p) were quantitatively assessed in six patients during clinically apparent grade II-III acute GVHD. Frequencies of responding Th p ranged from 1/13,000 to 1 4,000. The presence of host-specific Th-p was significantly correlated with the development of grade II-III acute GVHD (P = .0003 by Fisher's exact test). The detectability of host-specific Th-p preceded the clinical onset of grade II-III acute GVHD. Host-specific Th-p were no longer detectable after the clinical resolution of grade II-III acute GVHD. No subsequent chronic GVHD was observed in these patients. However, prolonged occurrence of host-specific Th-p was accompanied by clinically persisting acute GVHD and the onset of secondary chronic GVHD. In patients with no acute GVHD (grade 0) (n = 7) and grade I (n = 1) acute GVHD, host-specific Th-p were not detectable at all. We conclude that host-reactive Th are critically involved in the development and maintenance of acute GVHD and may contribute to the establishment of tolerance after genotypically HLA-identical sibling BMT. PMID- 8427977 TI - Acute graft-versus-host disease prophylaxis with methotrexate and cyclosporine after busulfan and cyclophosphamide in patients with hematologic malignancies. AB - The combination of two powerful immunosuppressive agents, methotrexate (MTX) and cyclosporine (CSP), has resulted in a significant decrease in the morbidity and mortality after allogeneic bone marrow transplantation (BMT). However, the additive toxicities from ablative preparative regimens may lead to suboptimal use of this combined immunoprophylaxis. We evaluated the efficacy and feasibility of administering MTX/CSP with busulfan (4 mg/kg/d for 4 days) and cyclophosphamide (50 mg/kg/d for 4 days) (BuCy4) in 101 consecutive patients with hematologic malignancies categorized into high- and low-risk groups receiving HLA-matched marrow grafts. Postgrafting immunosuppression consisted of MTX short course (15 mg/m2 on day 1 and 10 mg/m2 on days 3, 6, and 11) and intravenous CSP (1.5 mg/kg every 12 hours). Eighty-three patients (82.1%) received 100% of MTX calculated dose and 87 (86.1%) achieved a CSP therapeutic level (250 to 600 ng/mL) within a median of 16 days. Seventy-three patients (72.2%) received optimal immunosuppressive therapy comprising a full MTX course and achieving CSP therapeutic concentrations. The Kaplan-Meier estimated incidence of grade II to IV acute graft-versus-host disease (GVHD) was 9.2% for all patients and 5.5% in patients receiving optimal GVHD prophylaxis. Eighty-nine patients (88.2%) survived > or = 100 days posttransplant and 43 (48.3%) developed chronic GVHD, the majority of which were de novo (31 of 43). The estimated incidence of relapse was 28.9% for all patients and 14.8% in the low-risk group, with a median follow up of 24.5 months. High-risk features and the absence of chronic GVHD were significantly associated with relapse (P = .002 and .036, respectively) in multivariate analyses. Projected disease-free survival at 2 years was 52.3% for all patients and 65.2% in low-risk patients. Disease-free survival was significantly improved in optimally treated patients (P = .03) due to a lower incidence of early deaths from acute GVHD and infectious episodes. In conclusion, optimal delivery of MTX/CSP in association with BuCy4 resulted in a near complete abrogation of acute GVHD in HLA-matched transplants and a significantly improved disease-free survival. PMID- 8427978 TI - Rapid detection of a 13.4-kb deletion causing delta beta thalassemia in an Egyptian family by polymerase chain reaction. PMID- 8427979 TI - Human immunodeficiency virus detection and differential leukocyte counts are accurate and safer with formaldehyde-fixed blood. PMID- 8427980 TI - Long-term follow-up of 103 patients who received recombinant human granulocyte macrophage colony-stimulating factor after unrelated donor bone marrow transplantation. PMID- 8427981 TI - Plasma cell acid phosphatase in multiple myeloma. PMID- 8427982 TI - 9-cis-retinoic acid: effects on normal and leukemic hematopoiesis in vitro. AB - Retinoic acid exhibits effects on the proliferation and differentiation of many hematopoietic cells. Cellular responsiveness to retinoic acid (RA) is conferred through two distinct classes of nuclear receptors, the RA receptors (RARs) and the retinoid X receptors (RXRs). The RARs bind to both 9-cis- and all-trans-RAs, but 9-cis-RA alone directly binds and activates RXR. This suggested that 9-cis-RA could have expanded hematopoietic activities as compared with all-trans-RA. We compared the abilities of 9-cis- and all-trans-RAs to induce differentiation and inhibit proliferation of three acute myelogenous leukemia (AML) cell lines and fresh leukemic cells from 28 patients and found that: (1) 9-cis-RA in general was more potent than all-trans-RA in suppressing the clonal growth of two AML cell lines and 17 AML samples from patients, including four from individuals with acute promyelocytic leukemia (APL). Eleven leukemic samples, including three from patients with chronic myelogenous or chronic myelomonocytic leukemia, were relatively refractory to both retinoids. (2) The range of activities of both retinoids was similar except that the clonal growth of samples from three AML patients were inhibited by 9-cis-RA, but not by all-trans-RA. (3) Both retinoids inhibited the clonal proliferation of leukemia cells without necessarily inducing their differentiation; in fact, the only fresh AML cells that were able to undergo differentiation were from patients with APL and one individual with M2 AML. (4) Both retinoids enhanced myeloid and erythroid clonal growth from normal individuals, and 9-cis-RA showed slightly more stimulation of the myeloid clonal growth than did the all-trans-RA. Our study suggests that 9-cis-RA is worthy of further study for the treatment of selected individuals with AML. PMID- 8427983 TI - Cell-type-specific transactivation of the parathyroid hormone-related protein gene promoter by the human T-cell leukemia virus type I (HTLV-I) tax and HTLV-II tax proteins. AB - The human T-cell leukemia virus type I (HTLV-I) and HTLV-II Tax proteins are potent transactivators of viral and cellular gene expression. Using deletion mutants, the downstream parathyroid hormone-related protein (PTHrP) promoter is shown to be responsive to both HTLV-I and HTLV-II Tax as well as the AP1/c-jun proto-oncogene. Transactivation of PTHrP by Tax was seen in T cells but not in B cell lines or fibroblasts. A carboxy terminal Tax deletion mutant was deficient in transactivation of both the PTHrP and IL2R alpha promoters but not the HTLV-I long terminal repeat (LTR). Exogenous provision of NFkB rescued IL2R alpha expression but not the PTHrP promoter. Thus, HTLV-I Tax, HTLV-II Tax, and c-jun transactivate PTHrP and may contribute to the pathogenesis of hypercalcemia in adult T-cell leukemia. PMID- 8427984 TI - Prolonged survival of B-lineage acute lymphoblastic leukemia cells is accompanied by overexpression of bcl-2 protein. AB - Overexpression of bcl-2 delays the onset of apoptosis in lymphohematopoietic cells. We measured levels of bcl-2 protein in normal and leukemic human B-cell progenitors with a specific monoclonal antibody and flow cytometry. Normal immature B cells had low levels of bcl-2 protein; the intensity of fluorescence, expressed as molecules of soluble fluorochrome per cell, within CD10+ cells was 3,460 +/- 1,050 (mean +/- SD; 5 samples). In 16 cases of B-lineage acute lymphoblastic leukemia (ALL), cells had levels of bcl-2 that were strikingly higher than those of their normal counterparts (33,560 +/- 14,570; P < .001 by t test analysis). We next investigated whether the intensity of bcl-2 expression correlated with the resistance of immature B cells to in vitro culture. In 12 cases of B-lineage ALL, the cells recovered after 7 days of culture on allogeneic bone marrow stromal layers were 69% to 178% (median, 95.5%) of those originally seeded. Prolonged survival of leukemic cells in vitro was observed even in the absence of stromal layers in 6 of these 12 cases; the intensity of bcl-2 protein expression in these cases was 45,000 +/- 13,270, compared with 21,500 +/- 7,260 in the 6 cases in which greater than 99.5% of cells rapidly died by apoptosis under the same culture conditions (P = .003). Five immature B-cell lines, continuously growing in the absence of stroma, had the highest bcl-2 expression (79,400 +/- 20,330). By contrast, most normal CD19+, sIg-immature B cells died despite the presence of bone marrow stromal layers; 9.7% to 28.2% were recovered after 7 days of culture in three experiments. We conclude that abnormal bcl-2 gene expression influences the survival ability of B-cell progenitors. This may contribute to leukemogenesis and explain the aptitude of leukemic lymphoblasts to expand outside the bone marrow microenvironment. PMID- 8427985 TI - The 5q- syndrome: a single-institution study of 43 consecutive patients. AB - A favorable prognosis and a low rate of leukemic transformation has been attributed to the 5q- syndrome, a myelodysplastic syndrome (MDS) characterized by macrocytic anemia, hypolobulated micromegakaryocytic hyperplasia, and an interstitial deletion of chromosome 5. We examined the characteristics and outcome of 43 consecutive patients in our institution strictly defined by morphologic criteria and a solitary 5q- cytogenetic defect. The median age at diagnosis was 68 years, with a clear female predominance (7:3). Eighty percent of the patients were red blood cell transfusion-dependent at diagnosis and all untransfused patients had macrocytic indexes. In contrast, significant neutropenia or thrombocytopenia was rare. The French-American-British (FAB) class distributions were RA (72%), RARS (7%), RAEB (16%), and RAEB-IT (5%). At a median follow-up of 31 months, 56% of the patients survive, with a projected median survival of 63 months. The incidence of acute leukemia was 16% and was uniformly fatal. Clinical hemosiderosis occurred in 28% of the patients, resulting in two deaths. Neither survival nor the risk of leukemic transformation was predictable from initial clinical parameters, including FAB classification, Bournemouth score, and degree of aneuploidy. The lack of significant neutropenia and thrombocytopenia seemed to account for a very low incidence of infection and bleeding resulting in a prognosis equal or superior to historical patients with MDS. Therapeutic endeavors, including the use of corticosteroids, androgens, cis retinoic acid, pyridoxine, and danazol, were largely unsuccessful. PMID- 8427986 TI - Transferrin synthesis by mouse lymph node and peritoneal macrophages: iron content and effect on lymphocyte proliferation. AB - Transferrin is an essential requirement for lymphocyte proliferation, because it supplies activated lymphocytes with iron needed for cell proliferation. However, during inflammation or an immune response, the iron content of circulating transferrin, which is of hepatic origin, decreases. It is hypothesized that activated lymphocytes may therefore obtain transferrin-iron from an alternative source, and we have investigated the possibility that transferrin is synthesized locally in lymphoid tissues. It was found that lymph node cells from mice stimulated in vivo with Freund's complete adjuvant were able to synthesize transferrin, and this was because of the macrophage rather than the lymphocyte population. Transferrin synthesized by mouse lymph node or peritoneal macrophages contained iron and was able to promote mouse lymphocyte proliferation. Peritoneal macrophages activated in vivo synthesized more transferrin, released more transferrin-bound iron, and were more effective than resident macrophages at enhancing lymphocyte proliferation. These results suggest that transferrin synthesized by macrophages acts in a paracrine manner to support lymphocyte proliferation, thus eliminating possible detrimental effect of hypoferremia on the immune system. PMID- 8427987 TI - The distribution of erythrocyte phospholipids in hereditary spherocytosis demonstrates a minimal role for erythrocyte spectrin on phospholipid diffusion and asymmetry. AB - In the human erythrocyte membrane phosphatidylcholine and sphingomyelin reside mainly in the outer leaflet, whereas the aminophospholipids, phosphatidylethanolamine and phosphatidylserine, are mainly found in the inner leaflet. Maintenance of phospholipid asymmetry has been assumed to involve interactions between the aminophospholipids and the membrane skeleton, in particular spectrin. To investigate whether spectrin contributes to maintaining the phospholipid transbilayer distribution and kinetics of redistribution, we studied erythrocytes from hereditary spherocytosis patients whose spectrin levels ranged from 34% to 82% of normal. The phospholipid composition and the accessibility of membrane phospholipids to hydrolysis by phospholipases were in the normal range. Spin-labeled phosphatidylserine and phosphatidylethanolamine analogues that had been introduced into the outer leaflet were rapidly transported at 37 degrees C to the inner leaflet, whereas the redistribution of spin-labeled phosphatidylcholine was slower. The kinetics of transbilayer movement of these spin-labeled phospholipid in all samples was in the normal range and was not affected by the level of spectrin. Although these erythrocyte membranes contained as little as 34% of the normal level of spectrin and were characterized by several physical abnormalities, the composition, distribution, and transbilayer kinetics of the phospholipids were found to be normal. We therefore conclude that spectrin plays, at best, only a minor role in maintaining the distribution of erythrocyte membrane phospholipid. PMID- 8427988 TI - Quantitative assessment of erythropoiesis and functional classification of anemia based on measurements of serum transferrin receptor and erythropoietin. AB - We evaluated the quantitative value of a simple model of erythropoiesis, based on the basic assumptions that the red blood cell (RBC) mass determines erythropoietin (Epo) production, which in turn stimulates erythropoietic activity. The RBC mass was quantitated by direct isotopic measurement (RCM), Epo production by serum Epo levels, and erythropoiesis by the ferrokinetic measurement of the erythron transferrin uptake (ETU), the serum transferrin receptor (TfR) level, and the reticulocyte (retic) index, and was completed by an evaluation of overall marrow erythron cellularity. We studied a total of 195 subjects, including 31 normal individuals, 38 patients with polycythemia, and 126 patients with various forms of anemia. Instead of only quantitating Epo and erythropoiesis in absolute terms, we also evaluated them in relation to the degree of anemia or polycythemia, and expressed the results as a ratio of observed values to values predicted from the regression equations between hematocrit (Hct) on the one hand, and Epo, TfR, and ETU on the other, obtained in a carefully selected subpopulation. The slope of the regression of TfR (as well as ETU) versus Hct was very similar to the slope of the regression of Epo versus Hct. Average EPO and TfR (as well as ETU) values predicted from the regression equations were quite comparable to observed values in most groups of subjects, with exceptions predictable from knowledge of the pathophysiology of these hematologic disorders. We identified four major patterns of erythropoiesis, ie, normal, hyperdestruction (with variants of hemolysis or ineffective erythropoiesis), intrinsic marrow hypoproliferation, and defective Epo production. Dissecting out groups of patients showed much greater heterogeneity than when patients were analyzed by group. This was particularly true in the case of a hypoproliferative component being combined with hyperdestruction, giving what we called a "mixed disorder of erythropoiesis." We conclude that the pathophysiology of anemia can be assessed by a simple measurement of Hct, retic index, Epo, and TfR levels, with Epo and TfR being more informative when expressed in relation to the degree of anemia. The model is particularly useful for detecting the presence of multiple mechanisms of anemia in the same patient. However, it has limitations inherent to the relative invalidity of TfR in iron deficiency, the imprecision of a retic count, and the difficulty in distinguishing hemolysis from ineffective erythropoiesis in some patients and in recognizing a component of hyperdestruction in hypoproliferative anemia. PMID- 8427989 TI - Detection of hepatitis B virus in plasma using flow cytometric analyses of polymerase chain reaction-amplified DNA incorporating digoxigenin-11-dUTP. AB - Blood donations are routinely screened by multiple serologic assays for antigens/antibodies associated with infection by blood-borne viruses, including hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency viruses (HIV-1 and HIV-2), and human T-cell lymphotropic virus (HTLV-I and HTLV-II). A direct detection of these viruses would be more effective for the prevention of transfusion-transmitted infections than the indirect measurement of the variable host immune response to these agents. Because the polymerase chain reaction (PCR) for viral gene amplification offers the most sensitive and direct means of detecting viruses in blood, we have developed a nonisotopic PCR procedure for the detection of HBV, chosen as a prototype. The problems, common to previously described PCR methods, of nucleic acid extraction and inhibition of the PCR by plasma proteins were overcome by isolation of HBV from plasma by means of 450 microns polystyrene beads covalently coated with monoclonal antibody to the Pre S1 region of the viral envelope protein. Detergent lysis and proteinase K digestion of the immunocaptured virions isolated from plasma released the HBV DNA. A modified PCR-amplification protocol, incorporating digoxigenin-labeled dUTP in the amplified gene products followed by hybridization with a specific biotinylated oligonucleotide probe bound to streptavidin-coated 2.8-microns magnetic beads, allowed flow cytometric analyses of HBV-specific PCR products by means of antibodies to digoxigenin labeled with fluorescein isothiocyanate. The endpoint serial dilutions of pedigreed human plasma samples containing chimpanzee infectious dose (CID50) of 10(7) for adw and CID50 of 10(7.5) for the ayw subtypes were compared in repeated testing of PCR products by our immunoreactive bead (PCR-IRB) assay. HBV DNA was consistently detected in a 5 x 10(-10) dilution of each sample. In testing 20 coded specimens of blood donors, with or without serologic markers of HBV infection, the PCR-IRB was specific and more sensitive than the PCR analyses by slot blot hybridization with radioactive probe. The PCR IRB assay can be adapted for simultaneous detection of multiple blood-borne viruses by an automated flow cytometric analysis system. PMID- 8427990 TI - Long persistent bcr-abl positive transcript detected by polymerase chain reaction after marrow transplant for chronic myelogenous leukemia without clinical relapse: a study of 64 patients. AB - We report here the results of polymerase chain reaction (PCR) for bcr-abl transcript and clinical details derived from 64 chronic myelogenous leukemia (CML) patients after allogeneic bone marrow transplantation (BMT). A total of 139 samples (2 to 220 weeks after BMT) were analyzed and bcr-abl transcript was detected in 99 samples from 52 patients. Patients were defined as bcr-abl early negative (EN) if they had > or = 1 negative PCR result < or = 1 year post-BMT (n = 13), and bcr-abl late positive (LP) if they had > or = 1 positive PCR result > or = 1 year post-BMT (n = 21). Among LP patients, only two patients had hematologic/cytogenetic (clinical) relapse. Another 19 LP patients remained in clinical remission 7 to 130 weeks after positive analysis for bcr-abl transcript, including 5 patients who had persistent bcr-abl transcript detectable even 2 years after BMT. To estimate the relationship between clinical data and residual bcr-abl transcript, EN patients are compared with LP patients. However, no clinical data studied were significantly associated with the persistent PCR positivity. If only patients in chronic phase are compared, the t-test showed significant correlation between leukocyte count just before BMT and sustained bcr abl transcript (P < .05). These results suggest that PCR positivity is frequently observed in CML patients who sustain clinical remission after BMT, without being predictive of imminent clinical relapse. Tumor burden at the time of BMT may play an important role in the latency of bcr-abl positivity after BMT. PMID- 8427991 TI - Methotrexate, cyclosporine, or both to prevent graft-versus-host disease after HLA-identical sibling bone marrow transplants for early leukemia? AB - Optimal prophylaxis of graft-versus-host disease (GVHD) is controversial. We compared efficacy of three posttransplant immune suppressive regimens in 2,286 recipients of HLA-identical sibling bone marrow transplants for acute lymphoblastic leukemia (ALL) in first remission, acute myelogenous leukemia (AML) in first remission, or chronic myelogenous leukemia (CML) in first chronic phase. Six hundred forty received methotrexate, 977 received cyclosporine, and 669 received combined cyclosporine and methotrexate. In children, the three regimens resulted in similar outcomes. In adults, cyclosporine and methotrexate had comparable risks of acute and chronic GVHD. Compared with methotrexate, cyclosporine was associated with less interstitial pneumonia (relative risk [RR] = 0.6; P < .001), less treatment-related mortality (RR = 0.6; P < .001), more relapses (RR = 1.6; P < .05), and less treatment failure (relapse or death from any cause; RR = 0.7; P < .001). Different effects were observed in different leukemias. In ALL, the rate of leukemia relapse was increased with cyclosporine versus methotrexate, with no effect on other outcomes. In AML and CML, interstitial pneumonia, treatment-related mortality, and treatment failures were decreased with cyclosporine, with no increase in relapse. Similar analyses comparing cyclosporine plus methotrexate with cyclosporine alone showed that adults receiving the combination had less acute GVHD (RR = 0.5; P < .001), less chronic GVHD (RR = 0.7; P < .01), and less interstitial pneumonia (RR = 0.7; P < .001). Treatment failure (RR = 0.8; P < .05) was marginally reduced. Separate analyses in ALL and AML showed less acute GVHD with combined therapy, but no significant effect on other outcomes. In CML, acute GVHD, interstitial pneumonia, treatment-related mortality, and treatment failure were decreased with combined therapy. PMID- 8427992 TI - Bone marrow transplantation corrects the splenic reticuloendothelial dysfunction in sickle cell anemia. AB - In sickle cell anemia (SCA), the loss of reticuloendothelial function is the result of vasoocclusive events occurring in the spleen. Such asplenia occurs early in the course of the disease and is considered to be permanent in late childhood. In this report, three patients 10, 11, and 14 years of age suffering from severe SCA and found to be asplenic were treated by bone marrow transplantation (BMT). Before transplantation, all three patients had loss of reticuloendothelial splenic function, as assessed by the presence of abundant Howell-Jolly bodies on blood smears and absence of technetium 99m (99mTc) splenic uptake. After BMT, Howell-Jolly bodies disappeared from blood smear, whereas 99mTc isotopic scan found normal isotope uptake. Our data indicate that BMT can correct "permanent asplenia" in SCA patients. However, it remains to be determined if such treatment can also correct other SCA-related organ dysfunctions. PMID- 8427993 TI - Interleukin-10 and acquired immunodeficiency syndrome lymphomas. PMID- 8427994 TI - Tumor lysis syndrome provides evidence for dose-intense response for corticosteroids in prolymphocytic leukemia. PMID- 8427995 TI - Anemia of chronic disease. PMID- 8427996 TI - Detection of DNA rearrangements in the AML1 and ETO loci and of an AML1/ETO fusion mRNA in patients with t(8;21) acute myeloid leukemia. AB - The (8;21)(q22;q22) translocation is a frequent karyotypic abnormality seen in approximately 40% of patients with acute myeloid leukemia subtype M2 (AML-M2) and an abnormal karyotype. The translocation interrupts two genes, AML1 on chromosome 21 and ETO on chromosome 8, that are consequently fused in the der(8) chromosome to produce a novel chimeric gene and message. Selected genomic DNA probes from chromosome 21 and from chromosome 8 near the breakpoint junction detect rearrangements in the DNA of about 80% of the patients with the rearrangement at diagnosis and in relapse. We analyzed the DNA of 20 patients with t(8;21) AML by standard Southern blot with probes originating from chromosomes 21 and 8 near the breakpoint junction, and we identified rearranged bands in 17 of the 20 patients at diagnosis and in relapse. We also used the polymerase chain reaction (PCR) with appropriate primers from the AML1 and ETO genes to amplify the cDNAs from a cell line with the t(8;21) and from seven AML patients with the t(8;21). We detected a fused transcript in the cell line and in all of the patients analyzed, including three patients who did not show any rearrangement by Southern blot analysis and one patient in hematologic remission, who later relapsed. Combining the results from Southern blot and PCR analysis, we could detect the t(8;21) in all of the patients tested. These results indicate that, whereas several DNA probes used as genetic markers do detect the t(8;21) in most, but not all Southern blots of patients with AML, PCR amplification with primers from AML1 and ETO can be used as a more sensitive and accurate means for detecting this chromosomal abnormality, and for observing the patients' response to therapy. PMID- 8427997 TI - Interleukin-11 acts as an autocrine growth factor for human megakaryoblastic cell lines. AB - The cytokine interleukin-11 (IL-11) promotes normal human megakaryocytopoiesis in vitro. However, its role in abnormal megakaryocytopoiesis is not well known. Accordingly, we studied its effects on human megakaryoblastic cell lines CMK and Meg-J. IL-11 stimulated the proliferation of CMK and Meg-J in a dose-dependent manner with maximal growth being achieved at the concentration of 50 and 500 ng/mL, respectively. The growth of the cells was inhibited by anti-IL-11 antibody and IL-11 antisense oligonucleotides. IL-11 transcripts were detected in these two cell lines using a reverse transcriptase-polymerase chain reaction assay. These findings indicate that IL-11 might be an autocrine growth factor for megakaryoblastic cells. IL-11 transcripts also existed in other leukemia cell lines: HL-60, U937, and K562. However, the growth of these cells was not stimulated by IL-11, and was not inhibited by IL-11 antisense oligonucleotides. These results suggested that IL-11 might regulate malignant cells of the megakaryocytic lineage, in part by an autocrine loop. PMID- 8427998 TI - Recombinant human interleukin-11 stimulates megakaryocytopoiesis and increases peripheral platelets in normal and splenectomized mice. AB - The effects of recombinant human interleukin-11 (rhIL-11) on in vivo mouse megakaryocytopoeisis were examined. Normal C57Bl/6 mice and splenectomized C57Bl/6 mice were treated for 7 days with 150 micrograms/kg rhIL-11 administered subcutaneously. In normal mice, peripheral platelet counts were elevated compared with vehicle-treated controls after 3 days of rhIL-11 treatment and remained elevated until day 10. Splenectomized mice treated with rhIL-11 showed elevated peripheral platelet counts that were similar in magnitude to normal rhIL-11 treated mice. However, on day 10 the platelet counts in rhIL-11-treated, splenectomized mice were no longer elevated. Analysis of bone marrow megakaryocyte ploidy by two-color flow cytometry showed an increase, relative to controls, in the percentage of 32N megakaryocytes in both normal and splenectomized animals treated with rhIL-11. In normal mice, the number of spleen megakaryocyte colony-forming cells (MEG-CFC) were increased twofold to threefold relative to controls after 3 and 7 days of rhIL-11 treatment, whereas the number of bone marrow MEG-CFC were increased only on day 7. The number of MEG-CFC in the bone marrow of rhIL-11-treated, splenectomized mice was increased twofold compared with controls on both days 3 and 7 of the study. These data show that in vivo treatment of normal or splenectomized mice with rhIL-11 increased megakaryocyte progenitors, stimulated endoreplication of bone marrow megakaryocytes, and increased peripheral platelet counts. In addition, results in splenectomized mice showed that splenic hematopoiesis was not essential for the observed increases in peripheral platelets in response to rhIL-11 administration. PMID- 8427999 TI - Basic fibroblast growth factor counteracts the suppressive effect of transforming growth factor-beta 1 on human myeloid progenitor cells. AB - We have previously shown that basic fibroblast growth factor (bFGF) is mitogenic for human bone marrow stromal cells and enhances myelopoiesis in human long-term bone marrow culture. In the present study, we examined the mechanism by which bFGF enhances granulopoiesis. We observed that bFGF significantly abrogated the inhibitory effect of transforming growth factor-beta 1 (TGF-beta 1) on granulocyte-macrophage colony-stimulating factor (GM-CSF)-supported progenitor cell growth (P = .009). The partial reversal of TGF-beta 1-mediated suppression was dependent on the dose of bFGF used. In addition, we noted that the inclusion of neutralizing antibody to TGF-beta 1 significantly augmented the clonogenic response to GM-CSF. We have also shown that 10 ng/mL or 100 ng/mL of bFGF resulted in a 30% to 100% increase in GM-CSF-mediated progenitor cell growth (P = .0001). These data suggest that bFGF may enhance myelopoiesis by modulating the inhibitory response to TGF-beta 1. PMID- 8428000 TI - Interleukin-3 expression by activated T cells involves an inducible, T-cell specific factor and an octamer binding protein. AB - Interleukin-3 (IL-3) is exclusively expressed by activated T and natural killer cells, a function that is tightly controlled both in a lineage-specific and in a stimulation-dependent manner. We have investigated the protein binding characteristics and functional importance of the ACT-1-activating region of the IL-3 promoter. This region binds an inducible, T-cell-specific factor over its 5' end, a site that is necessary for the expression of IL-3 in the absence of other upstream elements. Over its 3' end, it binds a factor that is ubiquitously and constitutively expressed. This factor is Oct-1 or an immunologically related octamer-binding protein, and it plays a role in coordinating the activity of several regulatory elements. These characteristics make the ACT-1 site analogous to the activating ARRE-1 site in the IL-2 promoter. Furthermore, and despite a lack of sequence homology, the promoters of IL-3 and IL-2 share an organizational pattern of regulatory elements that is likely to be important for the T-cell specific expression of these genes. PMID- 8428001 TI - T- and B-lymphocyte differentiation potentials of spleen colony-forming cells. AB - Cells that generate splenic colonies within 8 days (day-8 colony-forming units spleen [CFU-s]) are generally thought to differentiate only into erythroid/myeloid cells. The T and B lymphocyte differentiation potentials of day 8 CFU-s were evaluated and compared with those of day-12 and 5-fluorouracil (5 FU) CFU-s. This was achieved by analyzing, after intravenous and intrathymic injection, the lymphocyte progeny of cells contained within individual splenic colonies collected at day 8 and day 12 post-bone marrow cell transfer into irradiated congenic recipients. A large majority of day-8 spleen colonies generated T cells when transferred intrathymically. After intravenous (IV) injection of day-8 colonies, donor-type thymocytes emerged in 33% of the animals reconstituted with only 1 day-8 colony, but in 83% of those inoculated with a pool of 5 colonies. All post-5-FU and 75% of day-12 colonies gave rise to thymocytes after IV injection. B cells were generated by a high proportion of day 8 colonies, and by all day-12 and post 5-FU colonies. These results demonstrate that progenitors of T and B lymphocytes are generated within spleen colonies produced by at least some day-8 CFU-s and virtually all day-12 CFU-s. Whether these progenitors are CFU-s themselves or committed precursors remains an open question. PMID- 8428002 TI - Red blood cell regeneration induced by subcutaneous recombinant erythropoietin: iron-deficient erythropoiesis in iron-replete subjects. AB - Limited red blood cell (RBC) regeneration often prevents collection of sufficient blood from autologous donors. We studied the effects of subcutaneous recombinant erythropoietin (rEPO) in subjects making frequent blood donations. Six healthy iron-replete male subjects took rEPO (200 U/kg) subcutaneously daily, and donated blood (450 mL) twice a week for 3 weeks. During a control study, these subjects also attempted twice-weekly blood donations without rEPO. Four other males given rEPO, including one with idiopathic hemochromatosis, waited until day 8 to begin blood donations. All healthy subjects took oral ferrous sulfate. Subcutaneous rEPO given with blood donations resulted in a marked reticulocytosis (mean peak value 568 +/- 159 x 10(9)/L v 235 +/- 77 x 10(9)/L, control study; P < .05), and enhanced RBC production at 28 days (1,208 +/- 227 mL v 719 +/- 161 mL, P < .05). rEPO in advance of blood donations was slightly less effective in normal subjects (941 +/- 139 mL, P < .05); however, the subject with hemochromatosis produced substantially more RBCs (1,764 mL) than any normal subject. rEPO-treated normal subjects (but not the rEPO-treated patient with hemochromatosis or untreated controls) produced iron-deficient RBCs with elevated zinc protoporphyrin levels and low hemoglobin content. These cells appeared within 1 week of rEPO administration and before laboratory confirmation of depleted iron stores. Thus, subcutaneous rEPO is an effective stimulant of erythropoiesis in nonanemic blood donors. However, in addition to eventual depletion of iron stores, early functional iron deficiency affects response to the drug. PMID- 8428003 TI - In vivo effects of recombinant interleukin-11 on myelopoiesis in mice. AB - Purified recombinant human interleukin-11 (rhuIL-11) was assessed for its in vivo effects on the proliferation and differentiation of hematopoietic progenitors as well as its capacity to accelerate the recovery of a drug-suppressed hematopoietic system. Dosage and time sequence studies demonstrated that administration of IL-11 to normal mice resulted in increases in absolute numbers of femoral marrow and splenic myeloid (granulocyte-macrophage colony-forming unit [CFU-GM], burst-forming unit-erythroid [BFU-E], CFU-granulocyte, erythroid, macrophage, megakaryocyte) progenitor cells and in stimulation of these progenitors to a higher cell cycling rate. This was associated with increased numbers of circulating neutrophils. Administration of IL-11 to mice pretreated with cyclophosphamide decreased the time required to regain normal levels of neutrophil and platelet counts in peripheral blood. In addition, IL-11 accelerated reconstitution to normal range of myeloid progenitors from bone marrow and spleen of myelosuppressed mice. These data suggest that IL-11 may play an important role in the regulation of hematopoiesis, and the application of this novel cytokine may have clinical therapeutic benefits. PMID- 8428004 TI - The kinetics of plasminogen activation by thrombin-cleaved pro-urokinase and promotion of its activity by fibrin fragment E-2 and by tissue plasminogen activator. AB - Thrombin hydrolyzes the Arg156-Phe157 bond in pro-urokinase (pro-UK), two residues from the activation site, generating a two-chain form (thromb-UK) believed to have little activity and that is resistant to plasmin activation. The kinetic constants for thromb-UK against synthetic substrate (S2444) were found to be essentially identical to pro-UK. Against native plasminogen, thromb-UK had a lower Michaelis constant (KM) and a higher (2-fold) catalytic efficiency. However, this difference with pro-UK was nullified by carboxypeptidase B (CpB) treatment of thromb-UK to remove the C-terminal arginine on the A-chain. Plasminogen activation by thromb-UK was substantially promoted by fibrin fragment E-2 but not by other fibrin derivatives, a phenomenon previously observed with pro-UK. Similarly, clot lysis by thromb-UK was promoted by tissue plasminogen activator because their combined effect was synergistic. Fibrinogenolysis in plasma occurred at 80-fold the concentration of thromb-UK as pro-UK, reflecting the 90-fold greater plasmin resistance of thromb-UK. Addition of a CpB inhibitor to the plasma enhanced fibrinogenolysis by thromb-UK and pro-UK by approximately 16%, consistent with the promotion of both forms by certain C-terminal lysines. In conclusion, CpB-thromb-UK corresponds functionally to a plasmin resistant form of pro-UK, indicating that the catalytic site of the single-chain pro-UK is unaffected by thrombin cleavage. The effect of CpB indicates that the C-terminal Arg of thromb-UK slightly enhances its affinity for plasminogen. Thromb-UK has potential plasminogen-activating activity at surfaces where C-terminal lysines, functionally comparable to fragment E-2, are found. PMID- 8428005 TI - Increased expression of platelet IgG Fc receptors in immune heparin-induced thrombocytopenia. AB - Our previous finding that heparin-dependent antibodies in heparin-induced thrombocytopenia (HIT) bind to platelets via platelet IgG Fc receptors (FcRs) prompted this study. Platelet FcRs in 16 patients with HIT, 23 control patients, and 42 normal subjects were studied. Patients with HIT had substantially increased platelet FcRs during the acute illness. Those who suffered serious thrombotic complications or died shortly after diagnosis had significantly more FcRs per platelet than those with milder disease. Consistent with their increased FcRs, platelets of patients with HIT showed increased aggregation reactivity to aggregated IgG and heparin-dependent antibodies. Platelet FcRs in patients with HIT remained elevated for 1 to 3 months after the acute illness then stabilized to a mean value not significantly different from either control group. The increased expression of FcRs on HIT platelets and their increased reactivity to heparin-dependent antibodies may contribute to the pathogenesis of thrombocytopenia and thrombosis in HIT. PMID- 8428006 TI - Defective signal transduction induced by thromboxane A2 in a patient with a mild bleeding disorder: impaired phospholipase C activation despite normal phospholipase A2 activation. AB - A patient with a mild bleeding disorder whose platelets responded defectively to thromboxane A2 (TXA2) was identified, and the mechanism of this dysfunction was analyzed. The platelets were defective in shape change, aggregation, and release reaction in response to synthetic TXA2 mimetic (STA2). When the platelet TXA2 receptor was examined with both a 125I-labeled derivative of a TXA2 receptor antagonist ([125I]-PTAOH) and [3H]-labeled TXA2 agonist ([3H]U-46619), the equilibrium dissociation rate constants (kd) and the maximal concentrations of binding sites (Bmax) of the platelets to both ligands were within normal ranges, suggesting that the binding capacity of their TXA2 receptor was normal. STA2 could not induce IP3 formation and intracellular Ca2+ mobilization, whereas these responses to thrombin were within normal ranges. GTPase activity was also decreased when the patient's platelet membrane was challenged with STA2. On the other hand, lysophosphatidylinositol formation, which is a direct indicator of phospholipase A2 (PLA2) activation, was found to be normal when the [3H]-inositol labeled platelets were challenged with STA2. Thromboxane B2 (TXB2) was also produced in response to STA2. These results suggested that the abnormality in these platelets was impaired coupling between TXA2 receptor and phospholipase C (PLC) activation. Furthermore, it is also suggested that the activation of PLA2 and PLC are separable events in thromboxane-induced platelet activation. PMID- 8428007 TI - It is the rare health care organization that has not implemented some type of quality program. PMID- 8428008 TI - Joint Commission nursing care standards: the framework for a comprehensive program to assess and improve quality. PMID- 8428009 TI - Bridging quality assurance and continuous quality improvement. PMID- 8428010 TI - Shifting gears: improving delivery of medications. PMID- 8428011 TI - Improving quality requires consumer input: using focus groups. PMID- 8428012 TI - Using patient perceptions to improve quality care. PMID- 8428013 TI - Nursing clinical pertinence review: a step toward quality improvement. PMID- 8428014 TI - Considerations on implementing CQI in rural hospitals. PMID- 8428015 TI - Medication use: indicators of safe practice. PMID- 8428016 TI - How are thresholds being used in quality assessment programs today? Any changes or updates? PMID- 8428017 TI - Decision-making model for a non-invasive diagnosis of compensated liver cirrhosis. AB - Liver biopsy is used as a gold standard in the diagnosis of chronic liver disease. However, this procedure is not without risk to the patients. This study was aimed to evaluate whether clinical, ultrasonographic, and biochemical variables may discriminate between well-compensated liver cirrhosis and non cirrhotic chronic liver disease. Logistic regression analysis was used to assess the independent predictive value of each variable. Moreover, the post-test probability of the diagnostic variables was converted into "weights" which positively correlated with the likelihood of diagnosis of liver cirrhosis. We applied a decisional rule based on the diagnostic "weight" of each variable to 412 patients, 278 with well-compensated liver cirrhosis and 134 with non cirrhotic chronic liver disease, diagnosed by liver biopsy with/without laparoscopy. By adding the diagnostic "weights" of each variable, liver cirrhosis and non-cirrhotic chronic liver disease were correctly diagnosed in 67% and 75% of cases, respectively. This result was validated in a split sample of 100 patients (50 with compensated liver cirrhosis and 50 with non-cirrhotic chronic liver disease) randomly selected from the acquisition sample. Based on this data, diagnosis of well-compensated liver cirrhosis can be reached in over 67% of cases without using invasive procedures. However, a prospective study is needed to confirm the clinical reliability of this method. PMID- 8428018 TI - Eco-Doppler evaluation of intestinal peristalsis in normal and in some pathological conditions: preliminary data. AB - Duplex-Doppler sonography could be employed in the quantitative investigation of intestinal motility. Preliminary data indicate reproductivity of the method in normal subjects and possible clinical applications in some pathological conditions affecting intestinal transit. Particularly, the possibility to discriminate between segments at different peristaltic activity seems to be very useful in intestinal obstruction. Further studies are necessary to validate this method. PMID- 8428019 TI - Multifocal leiomyosarcoma of the gastrointestinal tract requiring emergency surgical treatment for small bowel intussusception. AB - A case of multifocal leiomyosarcoma of the stomach, duodenum and jejunum presenting as an intestinal occlusion for small bowel intussusception is reported. Complete surgical excision of the tumour was not possible due to the wide dissemination. The patient died two months later for haemorrhage from an ulcerated gastric localization. Surgical and prognostic implications of disseminated gastrointestinal leiomyosarcoma are briefly discussed through a literature review. PMID- 8428020 TI - Junior staff experience with a decision aid for differential diagnosis of jaundice. AB - The Copenhagen pocket diagnostic chart can be used for early differentiation between obstructive and non-obstructive causes of jaundice. We present a reevaluation of the chart being used by a recently graduated physician and by two medical students. Non-error rates of 84-87% were found indicating that the students and the physician performed equally well. As to the observer error, the students and the physician typically differed on 0-2 of the chart's 21 items. This disagreement did not lead to a confident obstructive diagnosis being changed into a diagnosis of non-obstruction, or vice versa. PMID- 8428021 TI - Blastocystis hominis and blastocystosis (Zierdt-Garavelli disease). PMID- 8428022 TI - Amino acid transport systems. PMID- 8428023 TI - Stress proteins and the liver. PMID- 8428024 TI - Matrix-assisted laser desorption mass spectrometry of cytochromes P450. AB - Matrix-assisted laser desorption ionization on a time-of-flight (MALDI/TOF) mass spectrometer provides a rapid and accurate method for molecular weight determination of hydrophobic cytochrome P450 proteins (P450s). A mass accuracy of 0.075% was achieved by analysis of dialyzed P450 2B1 using bovine serum albumin (BSA) as an internal standard. The measured mass of cytochrome P450 2B2 with MALDI/TOF resulted in an average molecular weight which was higher than reported values. Immobilization procedures proved extremely effective on samples containing high concentrations of phosphate buffers. PMID- 8428025 TI - Matrix-assisted laser desorption of peptides and proteins on a quadrupole ion trap mass spectrometer. AB - The use of ultraviolet matrix-assisted laser desorption (MALD) to ionize peptides and proteins for analysis in a quadrupole ion trap is described. An ion source was modified to accommodate a fiber optic to transmit laser radiation from a nitrogen laser (337 nm) to the tip of the sample probe containing peptide or protein samples in a matrix of 2,5-dihydroxybenzoic acid (DHB) or 3,4-dimethoxy-4 hydroxy-cinnamic acid. Detection limits are demonstrated with 10 fmol of sperm whale myoglobin. The dimer of sperm-whale myoglobin was also observed at m/z 34,430. A comparison is made of the tandem mass spectrum of (MS/MS) of human angiotensin I desorbed by MALD to that of the peptide desorbed by liquid secondary-ion mass spectrometry. Both spectra were found to contain abundant structural information. PMID- 8428026 TI - The use of hydrogen-deuterium exchange to facilitate peptide sequencing by electrospray tandem mass spectrometry. AB - The utility of hydrogen-deuterium exchange for sequencing peptides by mass spectrometry is demonstrated. The number of exchangeable hydrogens in a peptide is readily obtained by electrospray analysis of the peptide dissolved in deuterated solvents. This information can be used, in conjunction with published computer algorithms for interpreting peptide mass spectra, to reduce significantly the number of candidate sequences that fit the experimental data. This information, when combined with fragment-ion information in the mass spectrum, greatly increases the reliability of sequence determination. PMID- 8428027 TI - Prevention of functional decline in older hospitalized patients. PMID- 8428028 TI - Alcohol use and alcohol problems among the elderly. PMID- 8428029 TI - Health promotion among elderly African-Americans. PMID- 8428030 TI - Exercise behavior among older adults. PMID- 8428032 TI - The language of epidemiology (XV): Biases in case-control studies. PMID- 8428031 TI - Why was there low participation in a health promotion and disease prevention program offered to Medicare beneficiaries? PMID- 8428033 TI - The status of health behavior indicators among older adults: focusing on the "high risk" elderly. PMID- 8428034 TI - Screening for older persons. AB - Our screening recommendations for older individuals are outlined in Figures 1 and 2. Our recommendations are parsimonious for two reasons. First, we are concerned about the potential harm of using unproven screening strategies. Second, when there are effective screening strategies, many of the target group are not being adequately screened. We agree with Clayman that it is much wiser to invest our scarce resources on improving the screening rates for clearly efficacious interventions before investing in strategies where efficacy is in question. PMID- 8428035 TI - Health and health care among Rhode Island's elderly. PMID- 8428036 TI - Primary prevention of functional decline in the elderly. PMID- 8428037 TI - Coexistence of morphea and systemic sclerosis. AB - While the association of morphea and systemic sclerosis (SSc) is considered to be a rare condition, we observed well-demarcated sclerotic skin changes indistinguishable from morphea in 9 of 135 SSc patients who visited our clinic during the last decade. We consider this rate of incidence (6.7%) to be high enough to consider morphea to be one of the skin involvements of SSc. There was a significantly (p < 0.01) higher incidence of morphea in males (3 of 5) than in females (6 of 130). Only 3 of 111 SSc patients positive for antinuclear antibody (ANA) also showed morphea, whereas 6 of 9 patients negative for ANA showed morphea (p < 0.01). Although the mechanism underlying the development of morphea in SSc patients remains unknown, our observations suggest a heterogeneous pathogenesis related to SSc gender and ANA type. PMID- 8428038 TI - Skin prick tests to common allergens in adult atopic eczema and rhinitis patients: reproducibility on duplicate and repeated testing. AB - Skin prick test reactivity to a battery of common allergens was examined in 26 adults with atopic eczema, in 9 patients with allergic rhinitis and in 10 nonatopic controls. In both patient groups, reactivity was most frequent for grass pollen, followed by rye, house dust mite, tree and herb, with no reactions to food allergens. Repeated tests at weekly intervals were less reproducible in weak reactions and in severely affected eczema patients compared to those lightly affected. Skin prick tests are therefore reliable and reproducible in atopics and provide means to counsel patients on avoidance of eliciting agents of their disease. PMID- 8428039 TI - Imbalance between plasminogen activator and its inhibitors in thiol-induced acantholysis. AB - Pemphigus may be an idiopathic disease or a syndrome induced by drugs, mainly thiol drugs. Autoantibodies, always present in the idiopathic form but often lacking in the drug-induced one, may cause acantholysis by activating endogenous proteolytic enzymes. Pathogenesis of drug-induced pemphigus when antibodies are absent has not been elucidated yet. Extracts of skin tissues cultured for 4 days with penicillamine, captopril or thiopronine were assayed for the presence of plasminogen activator (PA) and plasminogen activator inhibitors (PAI) on agar fibrin plates. Moreover, uPA, tPA, and PAI-1 were identified in the extracts by immunoenzymatic assay. The results have shown progressively decreasing amounts of PAI-1 in penicillamine, thiopronine and captopril-cultured tissue extracts, respectively. This suggests that the acantholytic potential of thiol drugs is directly correlated to their capability of reducing PAI-1 in the epidermal cells leading to increased PA activity. This PA-PAI imbalance may be itself the cause of intraepidermal splitting. PMID- 8428040 TI - Isotretinoin and acne in practice: a prospective analysis of 188 cases over 9 years. AB - A total of 188 acne patients (113 male and 75 female) with a mean age of 25 years (range 15-42 years) were treated with isotretinoin in doses ranging from 0.5 to 1 mg/kg. The study lasted 9 years. The treatment was not terminated until 2 months after total healing. The patients were then re-examined at regular intervals. In the event of a recurrence greater than grade 2 (pre-nodular threshold grade), they underwent a new course of treatment. At the end of the study, three groups were distinguished: (1) immediate, long-term, stable remissions following one single course of isotretinoin (111 patients with an average follow-up period of 27 months); (2) stable remissions following 2 or 3 courses of isotretinoin (54 patients with an average follow-up period of 16 months); (3) immediate partial remissions or partial remissions following several courses of treatment in 23 patients who continued to present with at least grade 3 acne. There was no significant statistical difference between the first two groups with respect to the age and sex of the patients and the grade and prior duration of the acne. The third group differed from the other two in having a greater proportion of patients with microcystic acne (p < 0.05) and women with endocrinological problems ((p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428041 TI - Diflorasone diacetate ointment 0.05% versus betamethasone dipropionate ointment 0.05% in moderate-severe plaque-type psoriasis. AB - We report the results of a 2-week double-blind, parallel clinical trial comparing two superpotent topical glucocorticosteroid ointments, diflorasone diacetate 0.05% and betamethasone dipropionate 0.05%, in psoriatic adults. Both corticosteroid ointments were fast acting and highly efficacious. 40 of the 44 patients who initially enrolled completed the trial. There were no statistically significant differences between the two glucocorticoids with respect to erythema, scaling, induration or the investigator's global evaluation after either 1 or 2 weeks of therapy. The level of patient satisfaction with the efficacy and cosmetic acceptability of these two category I glucocorticoids was similar. No systemic or local adverse reactions were noted. PMID- 8428042 TI - Dihydroxyacetone test as a substitute for the dansyl chloride test. AB - The hazards of the dansyl chloride test are not negligible. We introduce the dihydroxyacetone test as a safe substitute. Several advantages are highlighted including easy and reproducible measurements of the modifications in skin color by chromometry in the L* a* b* (luminance, hue, chroma) mode. The color variations in time as well as the fade-out of DHA-induced pigmentation by some cleansing skin care products are revealed 'in vivo' as well as 'ex vivo' on D Squame collections of stratum corneum. PMID- 8428043 TI - Melanoma associated with ring chromosome 7. AB - We report the case of a 17-year-old girl with a constitutional chromosome anomaly (ring chromosome 7), associated with multiple pigmented skin lesions, who developed malignant melanoma. Eight other cases of ring chromosome 7 are described in the literature; all present vascular or pigmented skin lesions, but our patient was the first to develop a malignant tumor. Genetic studies in malignant melanoma are discussed, as well as the possible link between a constitutional anomaly of chromosome 7 and the occurrence of malignant melanoma. This association is possibly not fortuitous, particularly since most ring chromosome 7 patients reported to date have pigmented skin lesions. PMID- 8428044 TI - Agminated blue nevi: case report and review of the literature. AB - Blue nevi may rarely appear in multiple form and grouped in a circumscribed area, a pattern of arrangement that is more properly designed under the term agminated blue nevi. In this paper a new case with light and ultrastructural studies is described, and the previously reported cases are reviewed. Histologically, there was a characteristic perifollicular arrangement of dermal melanocytes, most of which showed ultrastructurally an extracellular sheath. Agminated blue nevi seem to be benign lesions, but because of their rarity, no definite prognosis can be given. PMID- 8428045 TI - Epidermotropic metastasis coexisting with multiple primary cutaneous malignant melanomas. AB - A 45-year-old male, with a history of malignant melanoma on the thorax excised 4 years ago, presented a sudden eruption of more than 300 pigmented lesions. Fifteen excisions of tumors in different stages were carried out on four different dates. Dermatopathological examinations revealed features of primary as well as metastatic malignant melanomas. We suggest that besides the extensive dissemination there was also an activation of the resting melanocytes, that led to the formation of multiple primary melanomas. PMID- 8428046 TI - Staphylococcal scaled skin syndrome in an adult: possible influence of non steroidal anti-inflammatory drugs. AB - A 89-year-old woman presented with typical staphylococcal scaled skin syndrome (SSSS). An abscess of the left buttock related to non-steroidal anti-inflammatory drug (NSAID) injections was evacuated. We discuss the influence of NSAID administration on SSSS risk factors, i.e. renal insufficiency and immunosuppression. PMID- 8428047 TI - Bullous eruption with circulating pemphigus-like antibodies following interferon alpha therapy. AB - Two patients with classic Kaposi sarcoma developed bullous lesions and pemphigus like 'anti-intercellular-substance' antibodies when treated with interferon-alpha (IFN-alpha). In one of them direct immunofluorescence was consistent with pemphigus/pemphigoid features. The immunoprecipitation test showed no specific bands. The mechanism by which IFN-alpha may induce the production of anti intercellular-substance antibodies is discussed. PMID- 8428048 TI - Psoriasis and 2,3-biphosphoglycerate blood level. PMID- 8428049 TI - Longitudinal melanonychia associated with Bowen's disease: two new cases. PMID- 8428050 TI - Isotretinoin: 10 years on. PMID- 8428051 TI - Longitudinal melanonychia. PMID- 8428052 TI - Cutaneous findings in ring chromosome 7 syndrome. AB - All 9 cases of ring chromosome 7 syndrome already identified had skin lesions: 7 had vascular lesions, 5 showed large pigmented congenital nevi, 3 had cafe-au lait spots and in 1 case there were achromic spots on the trunk. One girl developed malignant melanoma at the age of 17 years. It appears that in patients presenting with growth retardation and pigmented skin lesions, with or without mental retardation, a cytogenetic study is indicated to search for ring chromosome 7, which is probably more frequent than suggested by the literature. PMID- 8428053 TI - Regressing nevoid nail melanosis in childhood. AB - The natural course of nail pigmentation was documented in 4 Japanese children. Although initial progression of melanosis concerning size and degree of pigmentation was noted, all cases showed a tendency to fade during the follow-up period. The published cases of nevoid nail melanosis in childhood were reviewed, and we concluded that they were essentially benign. However, should there be any suspicion of malignant changes, punch biopsy is indicated for medico-legal reasons. Management of growing pigmented nail streaks in childhood is one of the most difficult and delicate problems facing dermatologists, as they can be precursors of malignant melanoma. There is a condition of the nail known as longitudinal melanonychia or naevus striae unguis, possibly of nevoid nature. Prevalence rates of nevoid diseases showing pigmentation of the skin vary depending on the ethnic origin. In order to understand the natural course of nail area pigmentation in childhood, we have followed 4 Japanese children with such a condition. We report our observations and review the literature. PMID- 8428055 TI - Diagnostic value of exclamation mark hairs. AB - Exclamation mark (EM) hair can lead to the misdiagnosis of alopecia areata (AA) because it is widely considered as pathognomonic for AA. Typical EM hairs were also found in trichotillomania. EM hairs collected from 2 cases of trichotillomania and 9 cases of AA were compared under light microscopy. There were two sorts of EM hairs, one with frayed distal ends and the other with blunt distal ends. The majority (77.8%) of the EM hairs from AA had frayed distal ends, while the majority (82.2%) of that from trichotillomania had blunt distal ends. EM hairs, even with frayed distal ends, are not pathognomonic of AA. PMID- 8428054 TI - Study of sunbathing habits in children and adolescents: application to the prevention of melanoma. AB - Excessive sun exposure in the first 15 years of life has been shown to be a determinant risk factor for melanoma. This study was conducted on a randomly selected sample of 200 adolescents (13-14 years old) and 150 children (3 years old) in Marseille (South of France). Children and adolescents were examined and interviewed (mothers answered for young children). Our results show that a large number of highly sensitive children were not identified as such by their parents and most adolescents do not realize or at least admit being highly sun sensitive. Adequate sun protection measures were used in only 63% of 3-year-olds and 38% of adolescents. With reference to their constitutional skin sensitivity and taking into account their possible use of effective sun protection measures, 33% of the children and 62% of the adolescents were highly overexposed. Only good sun protection habits of the mother were predictive of acceptable sun exposure in children. In the adolescents the predictive variables were sun protection habits of the father and sunbathing only to obtain a tan. The main reason why adolescents sunbathed was embellishment. Conversely, most mothers said that they exposed their young children to the sun for health. Many adolescents and mothers were reasonably well informed but considered the risk of sun exposure to be exaggerated by the media. These results may be important to determine the targets of future melanoma prevention campaigns. PMID- 8428056 TI - Vascular dementia: a concept in flux. AB - Vascular dementia describes global cognitive decline attributed to the cumulative effects of ischemic vascular disease. Discrete and multiple cognitive skills, including memory, are successively lost as a result of focal cerebrovascular insults. Our understanding of vascular dementia has evolved over time, in part through our ability to neuroradiologically image changes in patients, and also through our increased understanding of the neuropsychologic organization of the brain. This paper reviews the development of our present understanding of vascular dementia, examines the status of "leukoaraiosis," pointing out current areas of controversy, and discusses clinical management. PMID- 8428057 TI - Role and expression of neurotrophins and the trk family of tyrosine kinase receptors in neural growth and rescue after injury. AB - Molecular cloning of genes for the neurotrophin family and the identification of their high-affinity receptors have recently contributed to our understanding of neurotrophic interactions in the vertebrate nervous system. From their primary sites of synthesis, novel neuronal populations that may be sensitive to the neurotrophins have been identified. Protective roles for these factors following epileptic, ischemic, and hypoglycemic insults have been inferred. Documented neurotrophic actions on basal forebrain cholinergic neurons and mesencephalic dopaminergic neurons imply future clinical applications for the treatment of dementia of both Alzheimer's and Parkinson's disease. Studies on structure function relationships of the neurotrophins hold promises for the development of specific receptor agonists and antagonists with possible basic science and clinical applications. PMID- 8428059 TI - Clinical neurochemistry. PMID- 8428060 TI - Cerebrovascular disease. PMID- 8428058 TI - Alzheimer's disease: new developments for noninvasive detection of early cases. PMID- 8428061 TI - Neuropsychology, dementia, and aging. PMID- 8428062 TI - Mitochondrial disorders in muscle. AB - The successful analysis of mutations in mitochondrial DNA has been a major step forward in our understanding of a number of baffling clinical syndromes, and the application of new technology to analyze and study mitochondrial DNA mutations has led to new insights into the pathogenesis of these disorders. Moreover, the field of mitochondrial DNA diseases is now expanding beyond the limited purview of relatively rare disorders to encompass clinical phenotypes that may be much more frequent in the population. PMID- 8428063 TI - Glial biology and disorders. AB - Much work has been devoted this year to the localization and mode of expression of growth factors and cytokines. Although it is not possible to extrapolate directly from in vitro to in vivo conditions, the plasticity of glial cells seems to be very influenced by growth factors. Astrocytes in vivo do not express many growth factors during normal conditions, but a pathologic event can lift these restrictions. Cytokines and their receptors have been localized on neuronal or glial cell types. The programmed cell death, well identified in neurons, seems to occur also in oligodendrocytes and may be influenced by survival factors. In the adult brain, glial progenitors are present and may be a potential source to generate myelinating oligodendrocytes for myelin repair. In the peripheral nervous system, axonal-Schwann cell signaling may function in both directions during development. Some animal neurologic mutants are models for human diseases; one of them, the Trembler mouse, has effectively led to the genetic characterization of Charcot-Marie-Tooth disease type 1a. As for myelin destruction, the relationship between demyelination and inflammation is still not very clear. PMID- 8428064 TI - Molecular genetics of neurodegenerative diseases. AB - Recent progress in human neurogenetics has led to the discovery of new modes of inheritance and disease expression, including 1) stably inherited duplications in Charcot-Marie-Tooth disease type 1a, 2) dynamic mutations in fragile X syndrome and myotonic dystrophy, and 3) identical mutations with different phenotypes in fatal familial insomnia and Creutzfeldt-Jakob disease. The mechanisms by which known mutations of the amyloid precursor protein lead to early-onset Alzheimer's disease remain unexplained, despite hundreds of recent studies of beta-amyloid. PMID- 8428065 TI - Treatment of acute cerebral infarction. AB - This review focuses on the recent advances in the management of acute ischemic stroke. We highlight the best current treatment, as proposed by different authors, as well as future lines of treatment. PMID- 8428066 TI - Antiplatelet and anticoagulant therapy. AB - Antiplatelet therapy is clearly indicated for long-term secondary prevention after transient ischemic attack and ischemic stroke. In stroke-free patients with atrial fibrillation, oral anticoagulants reduce the risk of stroke, and antiplatelet agents may be a lower risk alternative. For the early treatment of the acute phase of ischemic stroke, the role of antiplatelet and anticoagulant therapy is unclear, but is being evaluated in large clinical trials. PMID- 8428067 TI - The roles of heat shock proteins and immediate early genes in central nervous system normal function and pathology. AB - The expression and transcription of heat shock proteins and immediate early genes in the central nervous system are greatly affected by disease and injury. However, the participation of these genes in both normal and pathophysiologic function is uncertain. Experiments performed this past year shed light on the possible roles of heat shock proteins and immediate early genes in neurologic tissue. PMID- 8428068 TI - Strategies for preventing stroke. AB - The limited effectiveness of any therapy for acute stroke dictates that emphasis be placed on strategies for preventing stroke. The identification and management of risk factors is paramount. Modifiable risk factors include hypertension, smoking, hyperlipidemia, and diabetes. Patients with transient ischemic attacks or stroke are at risk for recurrent stroke and should be considered for antithrombotic therapy and carotid endarterectomy. Patients with nonvalvular atrial fibrillation should also be treated with antithrombotic therapy. The proper management of other cardiac, malignant, and hematologic disorders is important in preventing stroke in these high-risk patients. PMID- 8428069 TI - Surgery, angioplasty, and interventional neuroradiology. AB - This article discusses recent advances in the treatment of three cerebrovascular conditions, namely carotid stenosis, cerebral aneurysm, and arteriovenous malformation. In all three the aim of treatment is to prevent stroke, and in all three, recent advances in interventional neuroradiology are supplementing traditional surgical techniques. The benefits of endarterectomy for severe symptomatic carotid stenosis have been clearly demonstrated by recent trials, and early case series suggest that percutaneous transluminal angioplasty may provide an alternative treatment. Recent advances in the management of cerebral aneurysm include new treatments for cerebral vasospasm and the use of endovascular occlusion to treat inoperable aneurysms. Endovascular embolization for arteriovenous malformations also looks promising, but more randomized trials are required to establish the benefit of these interventional procedures. PMID- 8428070 TI - The economic approach to the stroke work-up. AB - Stroke is the leading cause of morbidity in the United States and the expenditure for stroke aftercare, including lost wages, is astronomical. Reduction of risk factors and use of the most accurate diagnostic technology allows for intervention prior to catastrophic neurologic deficit. The most advantageous combination of diagnostic testing with regard to risk-benefit has been debated, but it is generally agreed that the cost of even the most sophisticated stroke work-up is far less than that of stroke aftercare. PMID- 8428071 TI - Is stroke rehabilitation worthwhile? AB - Rehabilitation is an educational problem-solving process that focuses on disability and aims to minimize the patient's disability and distress and the stress on family members. There is established evidence that rehabilitation helps individual patients, and now there is good evidence that applying rehabilitation policies generally improves outcome in groups of patients. Moreover, well organized specialist rehabilitation is efficient because total resources are reduced. PMID- 8428072 TI - Neurobiology of schizophrenia. AB - Schizophrenia is the most prevalent of the major psychoses, but the underlying neurobiology of this debilitating disorder remains mysterious. Recent developments in molecular biology, neuroanatomic pathology, neurochemistry, and functional imaging suggest that a number of factors converge to produce schizophrenia. Specifically, an early neurodevelopmental "lesion," possibly within the mesial temporal lobe, may contribute to later temporolimbic-prefrontal dysfunction as the nervous system matures. Genetic factors appear to facilitate liability to schizophrenia, and dopaminergic and possibly other neurotransmitter systems may mediate clinical expression of the illness through newly recognized receptor subtypes. PMID- 8428073 TI - Disorders of memory in humans. AB - Memory is composed of dissociable systems that accomplish different forms of learning and that are mediated by distinct neural networks. Knowledge about the psychological nature and neural basis of specific memory systems has come primarily from lesion analysis, but this year functional brain imaging has provided a new source of convergent evidence. Recent findings are reviewed that provide new information about a limbic-diencephalic system critical for learning new declarative information, a temporal lobe system that stores long-term semantic knowledge in a domain-specific fashion, an occipital system that subserves visuoperceptual implicit memory, and a fronto-thalamo-neostriatal system that mediates the executive component of working memory and that plays an important role in some forms of sensorimotor skill learning. PMID- 8428074 TI - MR imaging contrast media: the global potential. PMID- 8428075 TI - Contribution of oral magnetic particles in MR imaging of the abdomen with spin echo and gradient-echo sequences. AB - An open phase III clinical trial of the oral contrast agent OMP (oral magnetic particles) was performed in 35 patients undergoing abdominal magnetic resonance (MR) imaging at 1.5 T with axial spin-echo and gradient-echo sequences. The diagnostic efficacy of OMP was examined by comparing pre-and postcontrast images. Bowel loops and abdominal organs were more easily recognizable after OMP ingestion, and the general quality of the images was improved because of fewer bowel-related artifacts. The diagnostic value of the postcontrast abdominal MR examination was superior or equal to that of the precontrast study, and additional information was obtained in 44% of the cases. Postcontrast gradient echo sequences increased confidence in the MR examination in 18% of cases. OMP was well tolerated and increased the quality and amount of diagnostic information acquired during the examination. Gradient-echo imaging was found to be a useful complement, but the need for a reduction in susceptibility artifacts was apparent and indicates that a decrease in TE or the use of rapid spin-echo sequences might be advantageous. PMID- 8428076 TI - Liquid oral magnetic particles as a gastrointestinal contrast agent for MR imaging: efficacy in vivo. AB - Recent in vitro studies suggested there is an optimal range of concentration and viscosity for a liquid formulation of oral magnetic particles (WIN 39996) for magnetic resonance (MR) imaging of the gastrointestinal (GI) tract. To determine whether this formulation is also effective in vivo and whether differing viscosity and administration regimen affect GI distribution of the contrast agent, a range of concentrations of iron (75, 150, and 200 micrograms/mL) and viscosities (1, 150, and 600 cp) were imaged in dogs at 1.5 T with conventional spin-echo and fat-saturation pulse sequences. The effects of dose regimen (single vs divided dose) and subject position (supine vs right lateral decubitus) were also studied. The 75 and 200 micrograms/mL concentrations were unacceptable for MR imaging, while 150 micrograms/mL was effective. The GI distribution of the contrast agent was affected jointly by viscosity, subject position, and dose regimen. The 150 micrograms/mL formulation produced excellent GI contrast enhancement in vivo for both 150- and 600-cp viscosities. The choice of optimal viscosity may depend on the preferred administration regimen. PMID- 8428077 TI - Preliminary evaluation of iron phytate (inositol hexaphosphate) as a gastrointestinal MR contrast agent. AB - A simple, effective, safe, and well-tolerated contrast agent is needed as a bowel marker for magnetic resonance (MR) imaging. The authors tested a variety of foodstuffs admixed with ferric iron as potential gastrointestinal MR contrast agents. Phytate (inositol hexaphosphate) more than doubled the relaxivity of solutions of ferric iron. Because of the improved relaxivity of iron phytate, the concentration of iron could be reduced substantially relative to free ferric iron (eg, ferric chloride or ferric ammonium citrate). Imaging studies were performed in five volunteers to determine the optimal dose of iron phytate and in five additional volunteers to test its effectiveness. A 200 mg/L concentration of ferric iron with phytate functions as an effective gastrointestinal MR contrast agent for T1-weighted abdominal MR imaging, significantly improving bowel contrast (P < .01). Blood studies after contrast agent administration showed no appreciable increase in serum iron. Compared with standard chelate complexes that decrease the relaxivity of a given paramagnetic ion, phytate not only decreases the absorption of the iron but increases its relaxivity. PMID- 8428078 TI - Safety aspects and pharmacokinetics of inhaled aerosolized gadolinium. AB - This study evaluated the pharmacokinetics and potential toxicity of inhaled aerosolized gadopentetate dimeglumine. The pharmacokinetics were evaluated with in vitro relaxometry of the lungs, blood, urine, and liver before and for up to 36 hours after a 5-minute inhalation of aerosolized gadopentetate dimeglumine (0.25 mol/L). For assessment of potential toxicity, hemodynamic variables were monitored during and for 10 minutes after inhalation. Extravascular lung water was measured before and after exposure. Finally, the potential for tissue damage was evaluated histologically. Pulmonary clearance of aerosolized gadopentetate dimeglumine was monoexponential with a half-time of 2.16 hours. Aerosolized gadopentetate dimeglumine was excreted through the kidneys, as shown by the decrease in urinary T1. Renal elimination was completed by 30 hours. No acute hemodynamic effect, histologic change, or induction of edema was demonstrated. This study shows that inhalation of aerosolized gadopentetate dimeglumine is well tolerated in rats and favors the further evaluation of this administration route, including clinical trials. PMID- 8428079 TI - Improved cine MR imaging of left ventricular wall motion with gadopentetate dimeglumine. AB - The assessment of left ventricular wall motion with cine magnetic resonance imaging in the cardiac long axes is useful; however, image quality is limited by reduced signal intensity due to saturation of blood flowing predominantly in plane. Twenty subjects were studied to evaluate the effects of gadopentetate dimeglumine on contrast between blood and myocardium. Contrast improved significantly on contrast agent-enhanced images in the long axes, at both end systole (vertical axis, +62%, P < .0001; horizontal axis, +67%, P < .0001) and end-diastole (vertical axis, +50%, P < .0001; horizontal axis, +26%, P < .004). The increase in the short axis was less and significant only at end-systole (+25%, P < .0001). Subjective scoring of the cines also showed a significant and clinically useful improvement in the long axes (P < .0001 for both) on contrast enhanced images. Improved contrast was most evident for 10 minutes after injection, but persisted for up to 20 minutes at end-systole in the long axes. Gadopentetate dimeglumine is useful for cine imaging of left ventricular wall motion when saturation effects prevent adequate clinical assessment. PMID- 8428080 TI - Quantitative analysis of gadopentetate dimeglumine excreted in breast milk. AB - Specimens of breast milk were obtained from each breast in a lactating patient for a 24-hour period after a gadolinium-enhanced MR imaging examination was performed. A quantitative analysis of gadolinium content in specimens obtained at 2, 11, 17, and 24 hours after the intravenous administration of gadopentetate dimeglumine demonstrated that gadolinium is excreted in small amounts (total of 1.60 mumol during a 24-hour period) and in similar amounts from each breast. These results suggest that a waiting period of 24 hours, with active expression of breast milk from each breast, should provide a reasonable safety margin for allowing resumption of breast-feeding. Additional data will be required before establishing definitive recommendations. PMID- 8428081 TI - Basic physics of MR contrast agents and maximization of image contrast. AB - This review summarizes the physical basis of magnetic resonance (MR) imaging contrast agents, including both T1 agents, such as gadolinium-DTPA (diethylenetriaminepentaacetic acid), and T2 or T2* agents, such as superparamagnetic iron oxides. The maximization of image contrast and lesion visibility with contrast agents is described, and the use of contrast agents in MR angiography and perfusion imaging is discussed. PMID- 8428082 TI - Proton relaxation enhancement. AB - Paramagnetic and superparamagnetic substances are used as contrast agents to enhance proton relaxation in magnetic resonance imaging. This review summarizes the physics of contrast agents, specifically the mechanisms by which contrast agents enhance T1 and T2 relaxation. The purpose is to provide a background for understanding the behavior of existing contrast agents in basic experimental and clinical studies. Terms such as magnetic dipole, dipole moment, magnetic susceptibility, diamagnetism, paramagnetism, superparamagnetism, and ferromagnetism are introduced. Two important interactions between the magnetic dipole moments of paramagnetic substances and the dipole moments associated with protons are described. The Solomon-Bloembergen-Morgan equations and other basic relaxation theory that has been confirmed experimentally are introduced to account for the dependence of relaxation on such parameters as the Larmor frequency, magnetic moment, accessibility of water molecules to the core of a contrast agent, and frequency of molecular motions. PMID- 8428083 TI - Biodistribution and toxicity of MR imaging contrast media. PMID- 8428084 TI - Chelates of gadolinium and dysprosium as contrast agents for MR imaging. PMID- 8428085 TI - Soluble-type hepatobiliary contrast agents for MR imaging. PMID- 8428086 TI - Iron oxides as MR imaging contrast agents. PMID- 8428087 TI - Status of liposomes as MR contrast agents. AB - Recent work on the development of liposomal magnetic resonance (MR) contrast agents has yielded structures with higher overall relaxivity than that of other nanoparticles of similar diameter. Liposomes incorporating membrane-bound complexes of manganase ("memsomes") produce greater hepatic enhancement per micromole of metal ion than either ferrite particles or paramagnetic chelates. Memsomes also hold promise for targeting of sites outside the liver. Work is in progress to take these agents into clinical trials. PMID- 8428088 TI - Use of MR imaging contrast agents in the brain. PMID- 8428089 TI - Dual effects of gadodiamide injection in depiction of the region of myocardial ischemia. AB - The high safety index of the nonionic magnetic resonance (MR) contrast agent gadodiamide injection permits a wide dose range without induction of hemodynamic effects. The wide dose range might confer a diagnostic advantage by providing both T1 enhancing and magnetic susceptibility effects for demarcating pathologic regions. The dual utility of this contrast agent for T1 and T2 enhancement in creating differential contrast between normal and acutely ischemic myocardium was explored. MR imaging was initiated 20-40 minutes after occlusion of the left coronary artery in two groups of rats: Group 1 (n = 8) received 0.3 mmol/kg gadodiamide injection before and after acquisition of T1-weighted images; group 2 (n = 10) received 0.5 mmol/kg before and after acquisition of T2-weighted images. Postcontrast T1-weighted images showed clear delineation of the ischemic region as a relatively low-signal-intensity area for 15 minutes. On postcontrast T2 weighted images, the ischemic region appeared as a relatively high-signal intensity area for 60 minutes. These effects were obtained with doses of gadodiamide injection that have safety indexes greater than those of gadopentetate dimeglumine. PMID- 8428090 TI - Application of contrast agents in MR imaging of the spine. PMID- 8428091 TI - Contrast-enhanced MR angiography. AB - Magnetic resonance (MR) angiography, although still in its infancy, is recognized as a valuable diagnostic tool. Investigation of the utility of contrast media as applied to MR angiography is, to no surprise, preliminary. In restricted instances, with present techniques, contrast media-enhanced MR angiography can provide additional valuable diagnostic information. Inspection of two-dimensional images (as opposed to three-dimensional projections) and comparison of MR images before and after administration of contrast agent are particularly important. Improved visualization of intracranial aneurysms, arteriovenous malformations, venous anomalies, and arterial occlusions has been demonstrated on three dimensional time-of-flight MR angiograms after intravenous administration of a gadolinium chelate, relative to studies performed before administration of the contrast agent. PMID- 8428092 TI - Contrast-enhanced perfusion-sensitive MR imaging in the diagnosis of cerebrovascular disorders. PMID- 8428093 TI - Use of gadolinium chelates in MR imaging of lesions of the extracranial head and neck. PMID- 8428094 TI - Contrast media for cardiothoracic MR imaging. AB - Contrast media have been used for the study of disease of the thorax. Most experience has consisted of the use of MR contrast media for improving the delineation of acute myocardial infarction, for demarcating the area of acute myocardial ischemia, and for demonstrating the area of potentially jeopardized myocardium after acute coronary occlusion. Contrast agents demonstrate the ischemic or jeopardized area as a zone of decreased signal intensity to normal myocardium. On the other hand, the magnetic susceptibility agents demonstrate the ischemic or jeopardized area as a zone of increased signal intensity to normal myocardium. Little experience exists concerning the use of MR contrast media for enhancing mediastinal and lung masses. The few reports to date demonstrate that contrast media can improve the conspicuity of thoracic masses. A new nonionic contrast medium, gadodiamide injection, has been shown to considerably increase S/N and C/N of thoracic masses. Further studies are necessary to define the role of MR contrast media in the evaluation of myocardial ischemia and thoracic masses in clinical practice. PMID- 8428095 TI - MR imaging of the breast. AB - The current strategy for breast cancer treatment involves early detection of the neoplasm before it has metastasized outside the breast, and surgical treatment of the lesion that minimizes deformity. Conventional methods of diagnostic imaging of the breast, including mammography, sonography, and galactography, do not adequately address clinical needs with regard to lesion characterization and staging. Magnetic resonance (MR) imaging has been proposed as a modality that may address these needs. The potential clinical uses of MR imaging are defined in terms of clinical needs and technologic requirements. Applications of MR imaging in detection of breast lesions can be divided into two major approaches based on the clinical questions to be answered: (a) improved specificity and (b) improved sensitivity. These approaches are defined by specific sets of clinical needs and have substantially different technologic requirements. The specificity approach is used to reduce the number of biopsies performed to confirm false-positive mammographic findings. The MR imaging device that is used to improve specificity must visualize only the lesions that are seen mammographically. The sensitivity approach requires the visualization of lesions not identified at mammography, so that breast disease can be more effectively staged for treatment. The technologic requirements for the sensitivity approach are considerably more rigorous because all lesions must be identified. The advantages and disadvantages of each of these approaches and the potential clinical ramifications are described. PMID- 8428096 TI - Abdominal imaging. PMID- 8428097 TI - Contrast-enhanced MR imaging of the female pelvis. PMID- 8428098 TI - Inflammatory joint disease: static and dynamic gadolinium-enhanced MR imaging. PMID- 8428099 TI - Effect of Gd-DTPA-BMA on magnetization transfer: application to rapid imaging of cardiac ischemia. AB - The addition of a paramagnetic contrast agent reduces the magnetization transfer effect between the free and restricted proton pools in both agar phantoms and cardiac muscle tissue. This reduction is due to the reduction in the intrinsic T1 of the free proton pool and increases the signal observed after a given magnetization transfer sequence. Images of ex vivo piglet hearts were obtained with a segmented snapshot FLASH (fast low-angle shot) sequence with a 128 x 128 matrix, four segments, and two signals averaged, resulting in an imaging time of 7 seconds. Magnetization transfer was induced by applying a DANTE (delays alternating with nutations for tailored excitations) pulse sequence in the intersegment interval. This was an efficient method of inducing magnetization transfer because it excites the restricted proton pool across the full frequency spectrum. Ischemia due to occlusion of the left anterior descending branch of the coronary artery could be visualized after infusion of gadolinium diethylenetriaminepentaacetic acid-bis(methylamide) (DTPA-BMA). Although the ischemia could be seen with the basic sequence, the contrast between ischemic and non-ischemic tissue improved when the magnetization transfer sequence was included. The most marked improvements in magnetization transfer were achieved with low doses of Gd-DTPA-BMA. PMID- 8428100 TI - Enhancement of tumor-liver contrast-to-noise ratio with gadobenate dimeglumine in MR imaging of rats. AB - The efficacy for tumor detection of the hepatocyte-specific contrast agent gadobenate dimeglumine (gadolinium-BOPTA/Dimeg) was evaluated in four different experimental tumor models in rats. Histologic findings were correlated with quantitative data derived from ex vivo relaxometry and in vivo magnetic resonance (MR) imaging. Noninfiltrating tumors showed maximal enhancement of liver parenchyma 5-10 minutes after contrast agent administration, with a plateau over the next 30 minutes. In contrast, infiltrating tumors, which caused hepatocellular injury and inflammatory changes, delayed maximal enhancement of tumor-free parenchyma by 15-20 minutes. Nonspecific tumor enhancement depended on tumor vascularity and occurred in the early phase after contrast agent administration. Despite differences in specific enhancement of tumor-free parenchyma and nonspecific tumor enhancement, tumor-liver contrast-to-noise ratios increased 96%-248% in all tumor models 30 minutes after intravenous administration of 75 mmol/kg Gd-BOPTA/Dimeg. Gd-BOPTA/Dimeg enhanced tumor conspicuity independently of the histologic characteristics of the tumor. PMID- 8428101 TI - In vivo relaxometry of three brain tumors in the rat: effect of Mn-TPPS, a tumor selective contrast agent. AB - T1 and T2 were determined simultaneously in vivo at 4.7 T in implanted rat brain tumors. Three different tumor cell lines were implanted in the right caudate nucleus: the F98 glioma, the E367 neuroblastoma, and the RN6 schwannoma. Their T1 and T2 values (mean +/- standard deviation [msec]), respectively, were 1,312 +/- 107 and 89 +/- 3 (glioma), 1,284 +/- 86 and 87 +/- 7 (neuroblastoma), and 1,338 +/- 85 and 86 +/- 9 (schwannoma). The T1 values (msec) of normal brain and muscle were 1,090 +/- 59 and 1,139 +/- 77, respectively, and the T2 values (msec) were 76 +/- 3 and 36 +/- 2, respectively. After injection of the contrast agent manganese (III) tetraphenylporphine sulfonate (TPPS) the T1 of all three tumors decreased by 30% and the T2 by 10%, whereas no such change in relaxivity was noted in normal brain. As a result, strong contrast enhancement of the three tumor types was seen on T1-weighted images. The tumor was clearly delineated and correlated with findings at histologic examination. This tumor enhancement was followed up for 4 days with quantitative relaxation time measurements, and the strong, selective reduction in T1 for all three tumor types after Mn-TPPS injection was preserved over the entire observation period. PMID- 8428102 TI - Tetra-p-aminophenylporphyrin conjugated with Gd-DTPA: tumor-specific contrast agent for MR imaging. AB - Tetra-p-aminophenylporphyrin (TPP) was conjugated with gadolinium diethylenetriaminepentaacetic acid (DTPA) and used as a contrast agent in magnetic resonance (MR) imaging to achieve tumor selectivity in nude mice. A substantial decrease in T1 was measured in excised tissues (kidneys, tumor, and liver) from mice that received the porphyrin derivative Gd2(DTPA)4 TPP. Toxicity and phototoxicity were less than those obtained with hematoporphyrin derivative in both L1210 lymphoblastic leukemia cells and HT 29 human colonic cancer cells, as determined with in vitro assays. MR images showed an enhancement of contrast between the tumor and adjacent tissue after injection of this agent. The results indicate that Gd2(DTPA)4TPP could be a useful prototype paramagnetic porphyrin MR imaging contrast agent with an affinity for tumors. PMID- 8428103 TI - MR imaging of hepatic iron overload in rat. AB - To investigate the relationship of hepatic signal intensity and T2 with histologic grading in an animal model of oral iron overload and to determine the duration of feeding necessary to produce abnormalities detectable on magnetic resonance (MR) images, hepatic iron overload was induced in 12 rats by feeding them a diet supplemented with 4% carbonyl iron for 2-11 weeks. Iron overload seen on MR images was graded independently and blindly by two radiologists as normal, mild, moderate, or severe. The rats were killed, and histologic findings were graded blindly by four pathologists using a similar subjective scale. Hepatic T2 values were estimated from spin-echo images. In the rats with iron overload, intracellular iron deposition was noted on histologic studies. On MR images, hepatic signal intensity and T2 decreased after only 2 weeks of dietary iron overload, and both continued to decrease with longer duration of feeding. There was significant correlation between iron overload duration and changes on MR images and between MR images and histologic grading (r = .92, P = .0001 for both). The mean T2 of hepatic iron overload decreased with longer duration of feeding. PMID- 8428104 TI - Comparison of Gd-EOB-DTPA and Gd-DTPA for contrast-enhanced MR imaging of liver tumors. AB - A new hepatobiliary contrast agent for magnetic resonance (MR) imaging, gadolinium-ethoxybenzyl (EOB)-diethylemetriaminepentaacetic acid (DTPA), was compared with Gd-DTPA to define the potential for improving tumor-liver contrast in a rodent liver adenocarcinoma model. With a T1-weighted spin-echo sequence, the contrast-to-noise ratio (C/N) for tumor before contrast agent administration was 5 (arbitrary units), the tumor appearing slightly hypo-intense with respect to liver parenchyma. After Gd-DTPA injection (0.1 mmol/kg), tumor enhanced more strongly than liver, resulting in an equalization of tumor and liver signal intensities and a decline in C/N to zero at 3 minutes after injection. After Gd EOB-DTPA injection (0.1 mmol/kg), liver enhanced more strongly than tumor. Five minutes after injection, C/N increased from 5 to 25 and remained above 17 for 50 minutes. The data indicate that Gd-EOB-DTPA yields higher and more prolonged tumor-liver contrast than Gd-DTPA on T1-weighted spin-echo images. The high liver tumor contrast after Gd-EOB-DTPA administration should prove clinically advantageous for MR imaging detection of focal hepatic masses. PMID- 8428105 TI - Chronic pancreatitis: MR imaging features before and after administration of gadopentetate dimeglumine. AB - Magnetic resonance (MR) imaging was performed in patients with a history (> 1 year) of inflammatory pancreatic disease. Calcification was seen at recent computed tomographic examinations in 13 patients and was not seen in nine patients. On fat-suppressed spin-echo images, the signal-to-noise ratio of the pancreas was significantly lower (P < .001) in patients with pancreatic calcification (18.2 +/- 2.5 vs 38.1 +/- 6.1). On fast low-angle shot images, the percentage of contrast enhancement was also significantly lower (P < .001) in patients with calcification (26.1% +/- 5.8 vs 78.7% +/- 15.9). The results suggest that MR imaging may be useful in evaluating patients with a long history of pancreatic disease for the presence of irreversible disease. PMID- 8428106 TI - First-pass evaluation of renal perfusion with TurboFLASH MR imaging and superparamagnetic iron oxide particles. AB - First-pass intrarenal hemodynamics were studied with superparamagnetic iron oxide particles and a T2-weighted TurboFLASH (fast low-angle shot) magnetic resonance (MR) imaging sequence. Four groups of five rabbits each were imaged after bolus injection of 40, 100, 140, and 200 mumol/kg iron, respectively. Images were acquired every 1.2 seconds, with an acquisition time of 700 msec. The signal intensity was measured in the cortex, outer medulla, inner medulla, and globally. In preliminary pathologic applications, two rabbits were imaged after ligation of the lumbar ureter and two after embolization of the renal artery. The reproducibility of the normal dynamics was evaluated with a cross-correlation test. On the images, the intravascular progression of the iron particles could be visualized within the cortex and the two compartments of the medulla in all cases. The maximal reduction in signal intensity in the cortex and medulla increased with the dose. The relationship between signal intensity decrease and dose was not linear, and the reproducibility of the signal intensity versus time plots was acceptable only at the 140 and 200 mumol/kg doses. The decrease in signal intensity was reduced and delayed in the embolized and hydronephrotic kidneys. PMID- 8428107 TI - Polymeric contrast agents for MR imaging of adrenal glands. AB - A variety of adrenal imaging agents have been used in nuclear medicine, but no agent has been developed for magnetic resonance (MR) imaging. The authors have previously observed accumulation of aminated macromolecules in adrenal glands. They now report the synthesis of a model polymeric aminated contrast agent for enhanced MR imaging of the adrenal glands. The model agent consisted of a poly-L lysine conjugate (molecular weight, 245 kd) that had 70% free epsilon amino groups and 30% diethylenetriaminepentaacetic acid (DTPA)-derivatized amino groups to bind indium-111 or gadolinium. One hour after intravenous administration of this compound, adrenal uptake was 10.1% +/- 0.7 of injected dose per gram of tissue. When all free epsilon amino groups of the polylysine were completely substituted with DTPA, adrenal uptake was 3.4 times lower, indicating the importance of free amino groups for adrenal uptake. MR imaging in rats showed that a dose of 0.08 mmol of gadolinium per kilogram of the agent was sufficient to enhance the signal intensity of adrenal glands. There hours after intravenous administration of the agent, signal intensity of the adrenal glands was 186% of precontrast values (liver, 165%; kidney, 91%). Fluorescence microscopy showed that the agent accumulated primarily in the cortical zona glomerulosa and in the adrenal medulla. These initial studies demonstrate the feasibility of designing contrast agents for MR imaging of the adrenal glands. PMID- 8428108 TI - Primary ovarian cancer: prospective comparison of contrast-enhanced CT and pre and postcontrast, fat-suppressed MR imaging, with histologic correlation. AB - Sixteen patients with clinically suspected malignant ovarian disease underwent contrast agent-enhanced computed tomography (CT) and magnetic resonance (MR) imaging in a prospective comparative study. MR imaging included fat-suppressed spin-echo and breath-hold FLASH (fast low-angle shot) before and after intravenous injection of gadopentetate dimeglumine. Histologic confirmation was obtained at laparotomy (n = 13) and biopsy (n = 3). Thirteen patients had histologically proven primary ovarian cancer. MR images showed the internal architecture of ovarian tumors better than CT in nine patients and equivalently in seven. MR images showed the relationship between ovarian tumors and adjacent pelvic structures (uterus [n = 9], sigmoid colon [n = 7], bladder [n = 7], and rectum [n = 3]) better than CT in nine patients and equivalently in seven. Intraabdominal extent of disease was better defined on MR than on CT images in nine patients, equivalently in six, and worse in one. Peritoneal metastases 1-2 cm in diameter were detected on MR images and missed on CT scans in six patients. In only one case did this result in a staging error with CT. The results suggest that MR imaging is at least equivalent and may be superior to CT in the evaluation of ovarian malignancy. PMID- 8428109 TI - Asbestos body and fiber concentrations in pathological autopsy tissues of patients with malignant peritoneal mesothelioma. PMID- 8428110 TI - Comparative analysis of emissions and diffusion of air PAHs at a coastal arid site (Patagonia, Argentina). PMID- 8428111 TI - Polychlorinated dibenzo-p-dioxins and dibenzofurans in rice straw smoke generated by laboratory burning experiments. PMID- 8428112 TI - Organic compounds found in Dokai Bay, Japan. PMID- 8428113 TI - Arsenic contamination levels in waters, soils, and sludges in southeast Spain. PMID- 8428114 TI - Synergistic effect of two- and four-component combinations of the polycyclic aromatic hydrocarbons: phenanthrene, anthracene, naphthalene and acenaphthene on Daphnia magna. PMID- 8428115 TI - Effect of triazine compounds on freshwater algae. PMID- 8428116 TI - Use of life-tables and application factors for evaluating chronic toxicity of kraft mill wastes on Daphnia magna. PMID- 8428117 TI - Ovarian effects of a sublethal concentration of mercuric chloride in the river frog, Rana heckscheri (Anura: Ranidae). PMID- 8428118 TI - Acute toxicity of technical captan to algae and fish. PMID- 8428119 TI - Carbofuran acute toxicity to freshwater algae and fish. PMID- 8428120 TI - Carbofuran acute toxicity to Eisenia foetida Savigny. Earthworms. PMID- 8428121 TI - Toxicity of produced water from crude oil terminals to Photobacterium phosphoreum, Chaetoceros sp., and Donax faba. PMID- 8428122 TI - Cadmium in the diet of the local population of Seville (Spain). PMID- 8428123 TI - Organochlorine pesticide and PCB congener content of French human milk. PMID- 8428124 TI - Microextraction of organophosphorous pesticides from environmental water and analysis by gas chromatography. PMID- 8428125 TI - Leaching losses of norflurazon through Mississippi River alluvial soil. PMID- 8428126 TI - Effect of acrylic polymer adjuvants on leaching of bromacil, diuron, norflurazon, and simazine in soil columns. PMID- 8428127 TI - Uptake of pre-emergent herbicides by corn: distribution in plants and soil. PMID- 8428128 TI - Should aging-associated cognitive decline be included in DSM-IV? PMID- 8428129 TI - Neuropsychological and event-related potential correlates of nonepileptic seizures. AB - A retrospective study of 14 patients with epileptic seizures and 11 with nonepileptic seizures, all taking antiepileptic drugs, found epileptic patients had significantly longer P160, N200, and P300 latencies on auditory event-related potential recordings. Patients with nonepileptic seizures had generally higher IQs and significantly greater psychopathology on neuropsychological scales. PMID- 8428130 TI - A pilot study of the neuropsychology of obsessive-compulsive disorder and Parkinson's disease: basal ganglia disorders. AB - This pilot study evaluated neuropsychological dysfunction in obsessive-compulsive disorder (OCD) and Parkinson's disease (PD). Both OCD and PD patients had impairment on visuoconstructional tasks but not on tasks of immediate memory and focused attention, suggesting common selective deficits in these two disorders with basal ganglia involvement. PMID- 8428131 TI - Schizophrenia, obsessive-compulsive disorder, and Tourette's syndrome: a case of triple comorbidity. PMID- 8428132 TI - Tourette's syndrome and nicotine withdrawal. PMID- 8428133 TI - Neuropsychiatric aspects of adult-onset Tay-Sachs disease: two case reports with several new findings. AB - Deficiency of hexosaminidase A causes the GM2 gangliosidosis known as Tay-Sachs disease. It is now known that this condition has several late-onset variants that cause numerous neuropsychiatric disturbances. Early recognition is important because treatment with phenothiazines and heterocyclic antidepressants may worsen the course. The authors report two cases with several new findings, including prominent psychiatric symptoms without psychosis early in the course of the illness. PMID- 8428135 TI - Focal paroxysmal EEG changes during atypical panic attacks. AB - Atypical panic attacks include features such as focal paresthesias or sensory distortions, but attempts to demonstrate a relationship to partial seizures have been unsuccessful. Two patients with atypical panic attacks had attacks during EEG monitoring: one during a routine EEG in the EEG laboratory, the other at home during ambulatory monitoring. Focal paroxysms of sharp wave activity appeared on both patients' EEGs coincident with the spontaneous onset of panic attack symptoms. Both patients remained conscious. The correlation of focal paroxysmal EEG changes with panic attack symptoms suggests that these attacks were produced by partial seizure activity. Further study of the relationship between panic attacks and seizures is indicated. PMID- 8428134 TI - Comparison between acute- and delayed-onset depression following traumatic brain injury. AB - Sixty-six patients admitted for the treatment of acute closed head injury were assessed for the presence of mood disorders during the in-hospital period and at 3-, 6-, and 12-month follow-ups. Diagnosis was made using a structured psychiatric interview and DSM-III criteria. A total of 28 patients had major depression at some time during the study: 17 had acute-onset depression and 11 had delayed-onset depression. Acute-onset depressions are related to lesion location and may have their etiology in biological responses of the injured brain, whereas delayed depressions may be mediated by psychosocial factors, suggesting psychological reaction as a possible mechanism. PMID- 8428136 TI - Reliability and applicability of movement disorder rating scales in the elderly. AB - Geriatric research clinicians were readily trained to use three scales initially developed for rating neuroleptic-induced movement disorders in mixed-age patients: the Abnormal Involuntary Movements Scale (AIMS), Simpson Extra Pyramidal Side Effect Scale (SEPSE), and Barnes Akathisia Scale (BAS). Interrater reliability, calculated by intraclass correlation coefficients, ranged from 0.79 to 0.93. Of elderly psychiatric inpatients tested, 97% were able to complete the examination, despite significant psychiatric, cognitive, and physical disabilities. AIMS, SEPSE, and BAS are reliable instruments for assessment of movement disorders in the elderly, representing an important first step in standardized evaluation of dyskinesia, parkinsonism, and akathisia in elderly psychiatric populations. PMID- 8428137 TI - Amnesic disorders. PMID- 8428138 TI - Age at disease onset and pattern of cognitive impairment in probable Alzheimer's disease. AB - Factor analysis of cognitive scores from 150 patients with Alzheimer's disease yielded two orthogonal factors: one (Factor 1) loading high on spontaneous speech, repetition, comprehension, reading, writing, digit span, and left/right discrimination; the other (Factor 2) loading high on long-term memory, orientation, object naming, and abstraction. Regression analysis, controlled for education and disease duration, showed Factor 1 scores to be lower in early-onset patients and Factor 2 scores to be lower in late-onset patients. These data corroborate results reported by previous investigators and suggest that Alzheimer's disease is age-dependent and heterogeneous in nature. PMID- 8428139 TI - A brief assessment of frontal and subcortical functions in dementia. AB - A brief assessment of cognitive and motor functions associated with the frontal/subcortical system was evaluated for discriminant validity. Patients with dementia of Huntington's disease and Parkinson's disease performed as well as normal control subjects on the Mini-Mental State Exam (MMSE) but significantly worse on the Frontal/Subcortical Assessment Battery (FSAB). Discriminant function analyses yielded significantly higher rates of accurate classification with FSAB and MMSE combined than with MMSE alone. Patients with Alzheimer's disease scored significantly lower than other groups on both measures. The authors recommend the FSAB as an adjunct to the MMSE for brief assessments of patients with suspected frontal or subcortical pathology. PMID- 8428140 TI - Diagnosing dementia: a comparison between a monodisciplinary and a multidisciplinary approach. AB - Discrepancies were examined in diagnostic outcome between a monodisciplinary approach and a multidisciplinary, criteria-based approach in patients referred to a university memory clinic. Of 278 patients not fulfilling dementia criteria, 19 had been previously diagnosed as demented (specificity: 0.93). In 60 of 152 demented patients, dementia had not been diagnosed before (sensitivity: 0.61). Underreporting was frequent for mildly demented patients and for patients with coexisting depressive symptoms. In patients referred by psychiatrists, sensitivity rates for dementia and Alzheimer's disease were low; in patients referred by neurologists, depression often went unreported. Results underscore the need for more frequent use of integrated multidisciplinary services for cognitively disturbed patients. PMID- 8428141 TI - Neuropsychological performance, mood, and complaints of cognitive and motor difficulties in individuals infected with the human immunodeficiency virus. AB - Seventy-nine military medical beneficiaries infected with human immunodeficiency virus (HIV+) and 27 HIV-seronegative control subjects (HIV-) completed a neuropsychological evaluation and a semistructured interview inquiring about difficulties in function. More HIV+ than HIV- subjects reported difficulties. HIV+ subjects reporting difficulties were significantly more likely to be deficient on attention, response speed, motor function, and memory than those not reporting difficulties. Findings for early-stage HIV+ subjects were similar. HIV+ individuals who complained of difficulties reported depression and anxiety symptoms significantly more frequently than those who did not complain, but these symptoms were not related to neuropsychological performance. Complaints of difficulties by HIV+ individuals may reflect either actual neuropsychological deficiency or mood disturbance, but the effects of each appear to be independent. PMID- 8428143 TI - Identification and management of radiculopathy. AB - Proper identification of compressive radiculopathy is essential before any treatment can be undertaken. The differential recognition of different pain patterns, sensory symptoms, and neurologic deficits provides the clinical guide to specific nerve root involvement. Appropriate radiology and imaging must correlate with symptoms and signs. Management includes surgical intervention when indicated for relief of radicular pain and restoration of function. PMID- 8428142 TI - Whole-blood serotonin and cognitive functioning in autistic individuals and their first-degree relatives. AB - The relationship between cognitive-intellectual abilities and whole-blood serotonin (5-hydroxytryptamine; 5-HT) in 18 autistic probands and their first degree relatives (n = 21 parents, n = 13 siblings) was investigated. Whole-blood 5-HT was significantly negatively associated with verbal-expressive/symbolic abilities for the entire sample. The proportion of variance in cognitive intellectual performances attributable to whole-blood 5-HT was substantial in the context of variance attributable to familial classification. The relationship between verbal-expressive abilities and whole-blood 5-HT, adjusted for race and familial classification, was noteworthy. Simple correlations between 5-HT and cognitive-intellectual performances were conducted. However, bias related to covariance attributable to race seriously limits such findings. The issue of bias and its relevance to previous research is discussed further. PMID- 8428144 TI - Lumbar disc disease. Microdiscectomy. AB - This article discusses a rationale for the use of microsurgical technique when treating lumbar disc herniations. The rigid surgical discipline of microlumbar discectomy is presented along with a suggested means to best preserve the future competence of the anulus fibrosis. PMID- 8428145 TI - Lumbar microdiscectomy with medial facetectomy. Techniques and analysis of results. AB - Lumbar microdiscectomy and medial facetectomy are safe and effective operative options in the treatment of symptomatic herniated lumbar discs in selected patients. We use this approach for patients with a lateralized intracanalicular disc herniation (rather than a broad-based disc rupture) without significant degenerative stenosis or a prior operation. The procedures can usually be performed easily and quickly with excellent illumination and visualization. The medial facetectomy component of the procedure allows early identification of the compressed nerve root and affords excellent exposure to the disc space without significant nerve root or thecal sac retraction. The results with microsurgical discectomy are good and the complications few. The limited incision and surgical dissection appear to minimize postoperative patient morbidity and may reduce the duration of the postoperative hospital stay. It is important that we not lose sight of the fact that the goal of lumbar disc surgery is to restore an individual's functional abilities and allow an expeditious return to daily activities as well as to work. Among carefully chosen patients, a skilled surgeon probably can achieve this goal using either standard or microsurgical techniques. PMID- 8428146 TI - Far lateral lumbar disc herniations. AB - Far lateral and related lumbar disc herniations that feature cephalad migration of fragments away from their disc space of origin can no longer be considered rare. Contemporary imaging techniques now reveal these herniations commonly. The anatomy of the foraminal and extraforaminal (far lateral) regions has been described and should be familiar to all spine surgeons. Each surgeon should be capable of recognizing the distinct entity of the far lateral disc herniation and should be adept at surgery in this region by one of several available techniques. I prefer a posterior, direct microsurgical approach through a midline incision featuring foraminotomy because of its ease, exposure, adaptability, and efficacy. PMID- 8428147 TI - Percutaneous discectomy for lumbar disc herniation. AB - Percutaneous discectomy for lumbar disc herniation is a well established, low risk procedure effective in 70% to 80% of appropriately selected patients. Advantages include the use of local anesthesia, minimal tissue disruption, no epidural fibrosis or scarring, and negligible biomechanical effect. The risk of automated percutaneous lumbar discectomy is significantly less than that of traditional surgery. PMID- 8428148 TI - Radiographic assessment of discogenic disease of the spine. AB - Magnetic resonance imaging has gained favor as the imaging modality of choice for evaluation of disc disease that affects the lumbar and thoracic spinal segments. This new noninvasive modality also competes favorably with myelography and CT for evaluation of cervical spine disc disease. An algorithmic approach to use of various imaging modalities for evaluation of suspected disc disease is provided in Figure 15. Important advantages of MR imaging relate to its multiplanar capability and unprecedented soft-tissue contrast for simultaneous evaluation of the thecal sac and spinal canal contents. These advantages often translate to more accurate and specific diagnoses related to degenerative disc disease. PMID- 8428149 TI - Instrumentation and fusion for discogenic disease of the lumbosacral spine. AB - A classification of lumbosacral discogenic pain is presented. Focus is placed on the pathophysiology, radiographic and clinical criteria, and role of fusion on the treatment of degenerative disc disease and segmental instability. Adjuncts, determinants, and techniques of fusion are discussed. Instrumentation systems, most applicable to the lumbosacral spine, are reviewed. PMID- 8428150 TI - Lumbar disc herniation in the pediatric patient. AB - Herniated lumbar discs are rare in teh pediatric population. The clinical features,diagnostic modalities, and treatment are discussed in detail.The overall surgical results have been excellent in more than 90% of patients in most series. The differences between children and adults with herniated lumbar discs are emphasized. PMID- 8428151 TI - Pregnancy and discogenic disease of the spine. AB - Low back pain is common during pregnancy and is of moderate or severe intensity in about one fourth of all pregnancies. The etiology is multifactorial but in most cases is related to the physical and physiologic changes brought about by pregnancy. For most women, the pain resolves spontaneously, although they remain at higher risk for increased LBP in future pregnancies and for the development of symptomatic disc disease in later life. Many of the common treatments for LBP are contraindicated or must be modified in this setting. Neurologically symptomatic herniated discs are rare during pregnancy, yet, when indicated, pregnant women can safely undergo surgery. PMID- 8428152 TI - Management of persistent or recurrent symptoms and signs in the postoperative lumbar disc patient. PMID- 8428153 TI - Complications of surgery for discogenic disease of the spine. AB - Disc disease is benign and not life threatening. Its long-term prognosis is uncertain, and spontaneous recoveries are common. The decision to perform surgery thus carries with it a heavy responsibility to provide safe treatment. "Good safe surgical technique" is essential, but it is much more than a well-rehearsed routine procedure. From preoperative evaluation to surgery to postoperative care, the surgeon must look ahead to problems that might arise. Where anticipated risks cannot be avoided, they should be minimized, and when adverse events occur, active recognition will lead to early and accurate management. PMID- 8428154 TI - Selection of patients for surgery. AB - Patient selection for lumbar laminectomy has been reviewed. The clinical decision for laminectomy depends on the mainstays of history, physical examination, and radiologic examination. A computer model to assist in this difficult task has been presented; the model is yet to be clinically tested. Psychological and psychosocial criteria play an important role in the decision-making process in these patients. PMID- 8428155 TI - Cervical disc herniations. The anterior approach to symptomatic interspace pathology. AB - The anterior operative approach to the cervical spine is an excellent and safe operative route for the treatment of symptomatic cervical disc herniations. The authors describe their surgical techniques and clinical results with this approach and discuss the issue of fusion following anterior cervical discectomy for spinal cord and cervical nerve root decompression. PMID- 8428156 TI - Surgical treatment of osteophytes and calcified discs of the cervical spine. AB - There are many options available for the surgical treatment of cervical spondylotic radiculopathy and cervical spondylotic myelopathy, with advantages and disadvantages to each. We have presented our approach to this clinical syndrome and the surgical techniques we prefer for each of its variants. Good, objective scientific data based on randomized, prospective clinical studies comparing the various surgical alternatives are lacking. The information that exists does not clearly favor any one single approach or operative option. It is critical to remember that each surgeon's experience and skill with a given technique have a profound impact on outcome. Contemporary spine surgeons should be well-versed in multiple operative techniques and should choose an appropriate approach for each individual patient based on that patient's clinical presentation and higher unique anatomy and pathologic condition, modified by the surgeon's personal experience. PMID- 8428157 TI - The posterior operative approach for cervical radiculopathy. AB - Cervical disc disease includes acute herniation and chronic disc degeneration with secondary changes in the associated bone. The latter may lead to the spectrum of cervical spondylitic stenosis, which is considered to be multilevel and may be more of a bony disease. Clinically, cervical disc disorders can be divided into several disorders. The disorder of true cervical radiculopathy is associated with lateral compression of the nerve root. When this condition is due to a lateral soft disc herniation or lateral bony stenosis, the posterior cervical laminoforaminotomy is commonly used. It is a procedure that works extremely well in the vast majority of patients and there is no risk of spinal instability; therefore, no fusion is required. The details of operative care have been described. In patients who have persistent radicular problems after a failed anterior cervical interspace procedure, the posterior cervical laminoforaminotomy with posterior wiring and fusion is a simple and effective operative option. PMID- 8428158 TI - Thoracic disc herniations. AB - Because of the protean nature of thoracic disc disease, surgeons should maintain a high order of suspicion of a thoracic disc herniation in the patient with unexplained localized back or torso pain and sensorimotor deficits. These patients should have MR imaging performed as a screening test, and, if suspicious for a thoracic disc herniation, confirmatory myelogram and postmyelogram CT imaging. Though the natural history is anecdotal, there appears to be a tendency for myelopathic symptoms and signs to be progressive, warranting surgical intervention. For radicular dysfunction or localized back pain, a conservative therapeutic plan is recommended. If intractable pain is demonstrated, and the diagnosis is certain, then surgical intervention is recommended. Once surgical intervention is recommended the surgical approach needs to be individualized according to the surgeon's skills and experience and the specifics of the patient's pathology. Appropriate surgical decision-making depends on an understanding of the variety of surgical options and their advantages and disadvantages, and an understanding of the biomechanical factors of the spine of the individual patient. Surgical concepts important to successful thoracic disc removal are (1) minimal spinal cord manipulation, (2) preservation of the neurovascular supply whenever possible, (3) minimal manipulation of the intercostal nerve, and (4) preservation of maximal bony and ligamentous attachments allowable for adequate exposure. Lastly it is recommended that the posterior longitudinal ligament be removed to ensure complete spinal cord decompression. PMID- 8428159 TI - Lumbar disc herniation. Standard approach. AB - This article describes precisely the standard surgical approach for the treatment of lumbar disc herniations. Each step in the procedure is presented in detail including anesthesia, positioning, surgical technique, and results. A brief introductory history of surgery for the ruptured intervertebral disc is also included. PMID- 8428160 TI - On the existence of suppressor cells. AB - Immunological suppression is one of the important aspects which govern the regulation of the immune response. Clonal deletion, clonal anergy and the activity of suppressor cells were proposed as mechanisms leading to this phenomenon. In this review we provide evidence for the existence of suppressor (Ts) cells and their role in autoimmunity in general and particularly in the system of experimental allergic encephalomyelitis (EAE). Ts cell lines and clones that downregulate in-vivo the autoimmune response are described. These results, as well as other recent advances in the understanding of Ts cells regulatory mechanisms and the molecular structures recognized by these cells, lead to the conclusion that Ts cells do exist and that they constitute a distinctive cell type in regard to their function and specificity. PMID- 8428161 TI - Interleukin-4 augments production of the third complement component by the alveolar epithelial cell line A549. AB - The elements of allergic inflammation and the involvement of helper T lymphocytes are increasingly being recognized in the immunopathogenesis of asthma. Allergen exposure leading to the activation of allergen-specific T cells present in the lung can result in the release of cytokines which in turn can locally stimulate the cellular constituents of the lung. The airway epithelial cells may be the key participants in such an interaction. Therefore, we examined the ability of T-cell derived IL-4 to modulate the production of C3 and C5 by the human type-II pneumocyte cell line A549, which is known to produce all the components and the regulatory proteins of the complement system. For estimation of C3 an ELISA detecting native C3 was used. Following stimulation of A549 with hrIL-4 a dose dependent (1-50 U/ml) enhancement of C3 production was observed, which reached its maximum (5-fold of unstimulated cells) at 48 h and gradually declined thereafter. Concentrations of hrIL-4 higher than 50 U/ml did not further increase C3 production. In parallel experiments hrIFN-gamma at concentrations between 10 and 50 U/ml stimulated the C3 production to more than twice the quiescent state level within 24 h. In the pneumocyte cell line A549 we demonstrated the expression of a gene for the IL-4 receptor which appears to mediate the biological effect of this lymphokine. A diminution in the functionally active C5, estimated by ELISA at the same time, was observed in supernatants of A549 cultures following stimulation with hrIL-4 as well as with hrIFN-gamma.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428162 TI - Antibodies to human serum albumin in familial dysautonomia. AB - Sera from familial dysautonomia patients are shown to contain high levels of antibodies to human serum albumin (HSA). The individual levels of anti-HSA antibodies correlate significantly with the levels of antibodies to bovine serum albumin and lactalbumin. These results and the findings that the anti-HSA level declines exponentially with age indicate that anti-HSA antibodies may be induced by aspiration of bovine milk antigens in infancy. The titer of the anti-HSA autoantibodies presently described (> 1:10,000) is much higher than those found in other diseases and may contribute to the pathological changes in familial dysautonomia. PMID- 8428163 TI - Polymyxin B-mediated lysis of tumor cells. AB - Polymyxin B (PmB) in the concentration range of 10-50 micrograms/ml is used routinely in immunological studies to neutralize low levels of contaminating lipopolysaccharide (LPS) in media or reagents. While using PmB for such a purpose unexpected results were obtained, which led to the finding that low levels of PmB are cytotoxic to certain tumor cells. Further examination of a panel of 10 tumor cell lines revealed that in an 18-hour 51Cr release assay, EL4 cells and EL4/ADM cells were very sensitive (lysed by > or = 10 micrograms PmB/ml), C1498 cells and REH cells were moderately sensitive (lysed by > or = 20 micrograms PmB/ml) and cells of the remaining 6 lines were resistant (lysed only by 100 micrograms/ml) to PmB. A similar pattern of sensitivity was observed when 3H-thymidine incorporation was used as a measure of PmB effects in cell proliferation. The PmB concentration needed to kill 50% of the tumor cells in a suspension differed greatly among lines; thus for cells of a resistant line 8-fold more PmB was required for 50% killing than for those of a sensitive line. PmB toxicity toward EL4 cells was shown to increase to a plateau level with increasing time of exposure; however, the higher the concentration the earlier the plateau was reached. LPS may prevent PmB toxic effects since PmB binds to the lipid A portion of the LPS molecule, but 100 micrograms LPS/ml was only able to reduce the toxicity of 10 and 20 micrograms PmB/ml, and not that of 50 or 100 micrograms PmB/ml.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428164 TI - Daily rhythmic variations in histamine in human blood. AB - We examined whether or not normal subjects have rhythmic changes of blood histamine levels. Daily predictable variations are present with 3 maxima and 3 minima and acrophase at 09.13. The significance of these changes is presently unknown. PMID- 8428165 TI - In vivo assessment of mast cell functional alteration induced by L-leucine methyl ester, using the passive cutaneous anaphylaxis technique. AB - We have recently reported that in vitro, mast cells were sensitive to the action of L-leucine methyl ester (Leu-OMe), a lysosomotropic compound. We now report the in vivo effect of Leu-OMe on mast cells, qualitatively assessed by using the passive cutaneous anaphylaxis (PCA) reaction. The L- but not the D-stereoisomer of Leu-OMe (25 mM) injected together with Dactylis glomerata, pollen-specific, IgE-containing serum inhibited the PCA reaction in rat skin triggered by a subsequent challenge with the corresponding allergen. When specific IgE antibodies were injected in the rat skin 3 days after the L-Leu-OMe, subsequent challenge with the antigen displayed a recovery of the PCA reaction. Thus, an in vivo L-Leu-OMe treatment, at a concentration which did not lead to any macroscopic tissue injury, elicited either an alteration of mast cell mediator release or an inhibition of allergen-induced vasoactive mediator release through a functional deactivation and/or a depletion of mast cells. PMID- 8428166 TI - Characterisation of dog allergens by means of immunoblotting. AB - Sera from 75 patients with clinical type I allergy against dogs were investigated by means of immunoblotting using extracts prepared from dog hair/dander (CAN XI D) and saliva. In addition, selected sera were tested on extracts made of hair, skin, salivary glands (parotis and submandibularis), serum and liver. A 69-kD IgE binding protein was identified in all extracts tested with an incidence of approximately 40% and shown to be dog albumin by means of inhibition experiments. In 96% of patients' sera IgE antibodies reactive with a 19-kD and/or a 23-kD protein of the hair/dander extract (CAN XI D) were observed. IgE binding to a 23 kD band was also detected in the hair and saliva extracts, but not in skin, salivary gland, serum and liver extracts. A 19-kD IgE-binding protein was strongly expressed in skin and to a lesser degree in saliva, but not in hair, serum and liver. Preincubation of patients sera with the hair extract and subsequent probing with the hair/dander extract (CAN XI D) inhibited IgE binding to the 23 kD protein whereas preincubation with the skin extract abolished IgE binding to the 19-kD protein. Using the hair/dander extract as inhibitor, IgE binding to the 19- and 23-kD proteins of saliva was abrogated. Thus it is concluded that the 23-kD protein is preferentially expressed in hair and saliva whereas the 19-kD protein is found in saliva and skin. Furthermore these two proteins are likely to represent immunologically independent major allergens. PMID- 8428167 TI - Isotypic analysis of grass pollen-specific immunoglobulins in human plasma. 1. Specialization of certain classes and subclasses in the immune response. AB - The specificity and isotypic profile of humoral immune responses to Dactylis glomerata (Cocksfoot) pollen was studied by isoelectric focusing (IEF) immunoprint analysis using 26 human plasma samples with high levels of Dactylis pollen-specific IgG4 (IgG4+ plasma) and 25 human plasma samples with low levels of specific IgG4 (normal plasma). Over 60 individual protein components in an aqueous pollen extract were separated by IEF and immunoprinted onto nitrocellose (NC). Following plasma incubation, bound IgE, IgG1-4, IgA1, IgA2 and IgM antibodies were detected on separate immunoprints with isotype-specific antibodies. Binding patterns of IgG4 and the majority of IgG1 and IgA2 antibodies in the IgG4+ plasma group very closely paralleled the binding patterns produced by the IgE antibodies from the same plasma and are described as the 'allergen repertoire'. In contrast, IgE, IgG4, IgG1 and IgA2 antibody reactivities to the 'allergen repertoire' were insignificant in the normal plasma group. These results suggest a qualitative, as well as a quantitative relationship between the immune responses which involve these 4 isotypes. Characteristic IgG2 and IgM antibody binding patterns, predominantly to non-allergenic antigens, were shared by the plasma from both groups, while IgG3 and IgA1 antibody binding patterns were highly variable from one plasma to another in both groups. One possible origin of the allergic diseases at the immunoglobulin heavy chain gene level is discussed. PMID- 8428168 TI - Allergenic cross-reactivity between Parthenium and ragweed pollen allergens. AB - Cross-reactivity of allergens from the pollen of the Compositae weeds, Parthenium hysterophorus (American feverfew) and Ambrosia (ragweed), in 2 groups of patients with different geographic distributions was studied. Parthenium-sensitive Indian patients, who were never exposed to ragweed, elicited positive skin reactions with ragweed pollen extracts. A significant correlation in the RAST scores of Parthenium and ragweed-specific IgE was observed with the sera of Parthenium and ragweed-sensitive Indian and US patients, respectively. RAST inhibition experiments demonstrated that the binding of IgE antibodies in the sera of ragweed-sensitive patients to short (W1) and giant (W3) ragweed allergen discs could be inhibited by up to 94% by Parthenium pollen extracts. Similar inhibition (up to 82%) was obtained when the sera of Parthenium rhinitis patients were incubated with ragweed allergen extracts. A dose-dependent proliferation of lymphocytes from a Parthenium-sensitive rhinitis patient with elevated levels of both Parthenium and ragweed-specific IgE was observed when incubated with Parthenium and ragweed pollen extracts. A 1.6-fold higher proliferation, however, was observed with Parthenium pollen extract at a concentration of 100 micrograms/ml. These results suggest that shared epitopes present on Parthenium and ragweed pollen allergens are recognized by both Indian and US patients sensitized by exposure to Parthenium and ragweed pollen, respectively. The high degree of cross-reactivity between Parthenium and ragweed pollen allergens suggests that individuals sensitized to Parthenium may develop type-I hypersensitivity reactions to ragweed and vice versa when they travel to regions infested with the weed to which they had not been previously exposed. PMID- 8428169 TI - Contra: CD8+ T-cell-mediated suppression without 'suppressor' T cells? AB - This commentary argues that conventional antiviral cytotoxic T cells act suppressively by eliminating antigen-presenting cells, sometimes including lymphocytes, T or B cells expressing relevant antigens of exogenous or endogenous, most commonly of viral origin. They thereby limit expression of antigens, and by eliminating the antigen-expressing cells stop immune responses. However, to argue that induced T cells (or antibodies) function to eliminate or neutralize antigen is the shortest summary of immunology and neither new nor heretic. PMID- 8428170 TI - Gastric cancer is curable in the West. PMID- 8428171 TI - A new hypothesis: iodine and gastric cancer. AB - The authors have hypothesized that iodine-deficiency (I-def) or in some cases iodine-excess (I-excess) is associated with the development of gastric cancer. They report a short review of their own work and general literature on this subject in three fields: (1) epidemiology, where geographical and temporal correlations between territories with I-def (or I-excess) endemic goitre and high GC-death rate are reported; (2) immunology, where the possible correlations between I-def, immune-deficiency and GC are reported; and (3) thyroid gland and stomach correlations, both being embryologically derived from primitive gut and able to concentrate iodine. This ability is impaired by nitrates, thiocyanate, salt and by I-excess, which in fact can cause goitre. In our study I-def goitrous people have shown more atrophic gastritis than normal subjects. These data enable us to hypothesize that I-def or I-excess might constitute a new risk factor for gastric cancer, both by regulating gastric trophism and by antagonizing the action of those I-inhibitors (such as nitrates, thiocyanate and salt) previously studied as risk factors for gastric cancer. PMID- 8428172 TI - Hypothesis on the relationship between gastric cancer and intragastric nitrosation: N-nitrosamines in gastric juice of subjects from a high-risk area for gastric cancer and the inhibition of N-nitrosamine formation by fruit juices. AB - The concentration of N-nitrosamines (NNA) in gastric juice was determined as an indicator of intragastric N-nitrosation in 85 subjects from a high-risk area for gastric cancer (GC) to examine the relationship between N-nitroso compounds (NOC), pH and intragastric lesions under strictly controlled conditions. Mean gastric pH in subjects with GC or dysplasia (Group GD, 5.0 +/- 2.7) was higher than that from subjects with intestinal metaplasia (Group IM, 3.8 +/- 2.1, p = 0.068) and significantly higher than in those with normal mucosa or superficial gastritis (Group NS, 2.6 +/- 1.9, p < 0.001). No significant difference (p > 0.1) was found in total NNA concentrations between the three groups (GD 1.81 +/- 1.05 micrograms/l, IM 1.46 +/- 0.79 micrograms/l, NS 1.56 +/- 1.38 micrograms/l). However, two obvious peaks of nitrosation were observed at pH ranges of < 2.0 and 5.5-7.5. These observations were confirmed by using the N-nitrosoproline test in the same subjects under the same conditions (r = 0.772, p < 0.05). These results indicate that intragastric nitrosation can occur in both acidic and nearly neutral conditions. The first peak is related to acid-catalysed nitrosation (ACN) and the second is related to biologically catalysed nitrosation (BCN). According to these and other published results the hypothesis that there are two basic mechanisms, ACN and BCN, for intragastric N-nitrosation in humans is explored. Gastric carcinogenesis in high-risk areas is more likely to be related to intragastric NOC formed by ACN, compared to low-risk areas where it is more likely to be related to intragastric NOC formed by BCN. Fruit juices and orange peel significantly inhibited intragastric nitrosation by both ACN and BCN. PMID- 8428173 TI - WHO initiative: coping with cancer in Europe. PMID- 8428174 TI - Enhancing mammography uptake: who do women listen to? AB - Previous work has established that women who attend for mammography differ from non-attenders in a number of socio-demographic and attitudinal characteristics. The present study was conducted to determine whether women who attended for mammography differed from non-attenders in a number of key areas: (1) in how they obtained information about screening; (2) in their understanding of the disease and basic screening precepts; and (3) in the extent to which they perceived their general practitioners (GPs) and other members of the primary case team, such as practice nurses, had actively promoted the programme. Three hundred attenders and 300 non-attenders were interviewed in their own homes using a structured questionnaire. Only 5% of women interviewed had ever asked their GP for any advice about breast screening, and only 18% recalled their family doctor every discussing or raising the subject with them. Although attenders and non-attenders differed significantly in their understanding of the scope and purpose of screening, both groups obtained information more often from friends and relatives and broadcast media than from official sources. Attenders were more likely to cite material in the GP's surgery as an important source of information (chi 2 = 5.1, p = 0.02). Attenders were marginally more likely than non-attenders to have previously attended a well-woman clinic in primary care (chi 2 = 3.1, p = 0.08) and were more likely to say that such clinics were being offered by their family doctor (chi 2 = 9.8, p = 0.008).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428175 TI - Handling antineoplastic drugs in the European Community countries. AB - Alkylating antineoplastic drugs (ADs) are carcinogenic to humans and most ADs cause reproductive failures in animal experiments. Studies of occupationally exposed health personnel have shown increased risks for miscarriages (two studies), malformations (two studies) and leukaemia (two studies). During the past decade, work practice guidelines have been issued by authorities in several countries but universal agreement on a set of guidelines which would secure both the work environment and the treatment of patients is yet to come. We reviewed the existing guidelines in the European Community (EC) countries on health personnel's handling of ADs. Five countries had guidelines on the safe handling of ADs. The main principles were to use exhaust cabinets and personal protective equipment. Two sets of guidelines included recommendations for pregnant women. In general, the EC countries' guidelines were less extensive than those issued by the US Occupational Safety and Health Administration. It is argued that the existence of more uniform guidelines in all the member states would be valuable. Specifically, measures should be devised to prevent adverse effects on reproduction. PMID- 8428176 TI - Malignant Barrett's oesophagus. AB - Barrett's oesophagus is a pre-malignant condition with an increased risk of adenocarcinoma. The prevalence of adenocarcinoma in Barrett's oesophagus is about 10% but its true incidence in the general population is unknown. The development of adenocarcinoma in Barrett's oesophagus is a multi-step process. Gastro oesophageal reflux symptoms are absent in many Barrett's patients and both Barrett's oesophagus and adenocarcinoma are usually but not always diagnosed simultaneously. When a carcinoma is identified, the treatment of choice is resection. Three-stage oesophagectomy is considered the most appropriate procedure. The prognosis of Barrett's carcinoma is dismal and the survival rate is related to stage of the tumour. However, encouraging results have been reported in the past 5 years. Endoscopic surveillance for Barrett's oesophagus is still a controversial topic but for some high-risk subgroups of patients regular surveillance is advocated. At the present time, dysplasia is the best available indicator of malignancy in Barrett's oesophagus. PMID- 8428177 TI - Oesophageal carcinoma: the need for screening. AB - Oesophageal cancer is a substantial cause of mortality in the Western world and recent data indicate that the incidence is increasing. Despite better understanding of the pathogenesis and in the surgical management of the disease, little improvement in the survival rates has been achieved anywhere in the world, especially because screening for detection of premalignant lesions cannot, at present, be adequately applied to populations at risk. The present review summarizes current knowledge of the use of conventional screening methods as well as possible applications of new techniques to targeted populations to permit earlier diagnosis of premalignant lesions of the oesophagus. PMID- 8428178 TI - Prevalence survey of precancerous lesions of the oesophagus in a high-risk population for oesophageal cancer in France. AB - The prevalence of precancerous lesions of the oesophagus and their association with alcohol drinking, tobacco smoking and some dietary factors were examined in an endoscopic survey carried out in Lower Normandy, France, a high-risk area for oesophageal cancer. The study included 134 male volunteers of 35-64 years of age. In 124 of the volunteers oesophageal biopsies were evaluable. At histology, the prevalence of chronic oesophagitis, epithelial atrophy and dysplasia was 63%, 1.6% and 4.8%, respectively. The prevalence of these precancerous lesions was significantly associated with cigarette smoking and frequent consumption of butter. PMID- 8428179 TI - Effect of beta-carotene supplementation on the activity of ornithine decarboxylase (ODC) in stomach mucosa of patients with chronic atrophic gastritis. AB - Increase of ornithine decarboxylase (ODC) activity is known to be associated with cell proliferation and, very likely, with tumour promotion. This prompted us to study the activity of ODC in gastric mucosa of patients with chronic atrophic gastritis that has been considered as a precursor of stomach cancer. Examination of 124 patients with this disease revealed the considerable increase in ODC activity in atrophic mucosa (29.8 +/- 2.9 vs 7.9 +/- 1.8 units in normal mucosa, p = 0.001). Supplementation of the patient's diet with beta-carotene (20 mg daily during 3 weeks) results in a statistically significant decrease in ODC activity in gastric mucosa. The data obtained confirm the possibility of application of ODC determination to the detection of early premalignant lesions and suggest the antipromoter activity of beta-carotene in gastric carcinogenesis. PMID- 8428180 TI - Effect of exercise on intestinal tumour development in the male Fischer rat after exposure to azoxymethane. AB - Thirty-two male Fischer rats were exercised in a treadmill, 2 km daily on weekdays for 38 weeks, after three subcutaneous injections of azoxymethane, 15 mg/kg body weight. Randomly selected control animals were housed in regular 38 x 22 cm cages in groups of no more than three rats per cage. All rats were allowed free access to food and water. The exercising rats ate about 20% more than the sedentary rats, but gained less weight. Fat stores were considerably smaller in the exercised rats whereas the lean body mass and ash content were similar. After 38 weeks significantly fewer rats in the exercise group had developed neoplasia of the colon mucosa. PMID- 8428181 TI - Supplemental dietary calcium and inhibition of colon cancer. PMID- 8428182 TI - Smoking and adult T-cell leukemia/lymphoma. PMID- 8428183 TI - Communication in the physician-patient relationship. AB - Political, legal, ethical, social, economic, and technological changes in the twentieth century have produced a profound effect on the health care and health status of Americans and the way in which physicians and patients communicate. In the latter half of this century, the responsibility for individual health care has shifted from a physician-oriented, paternalistic approach to a patient centered one. Patients now assume two identities: health consumers and active participants in the medical decision-making process. This phenomenon has created an environment where consumer demand for information has shifted from a single focus on symptoms, diagnosis, and treatment of diseases to an increasing preoccupation with cost, quality, and access to health care. This shift emphasizes the critical role played by medical librarians in the dissemination of needed information, and it challenges librarians to take a leadership role in opening newer channels of communication between physicians and patients. The 1992 Janet Doe Lecture analyzes the evolutionary change in the physician-patient relationship and its modes of communication, projects future roles for medical librarians, and provides an extensive list of references for further reading. PMID- 8428185 TI - Augmenting GRATEFUL MED with the UMLS Metathesaurus: an initial evaluation. AB - Clinicians in patient care settings must be able to locate relevant recent medical literature quickly. Computer literacy is increasing, but many clinicians remain ill at ease with search strategies for online bibliographic databases. As part of an ongoing project to simplify the translation of clinical questions into effective searches, a Unified Medical Language System (UMLS) Metathesaurus tool was designed. The authors compared bibliographic searches by relatively inexperienced users employing only GRATEFUL MED to searches done using GRATEFUL MED augmented with this tool. The users were clinicians examining questions related to a test set of clinical cases. Their problems and successes were monitored; the results suggest that the addition of a thesaurus helps resolve some problems in citation retrieval that trouble the novice user. By helping the user understand indexing terms in context and by reducing typing errors, a thesaurus can help provide an intelligent solution to lexical mismatches in bibliographic retrieval. PMID- 8428184 TI - Physicians' use of computer software in answering clinical questions. AB - Descriptive data about the use of medical information software were gathered from physicians who were early users of these resources. Eight clinically active internists and medical subspecialists were lent a microcomputer loaded with six commercially available medical information software products. Participants used the software for two weeks to answer questions arising in their practice and completed written questionnaires. They recorded a total of 50 questions (between 3 and 11 per participant per two-week study period). Using the workstation, participants answered 20 questions (40% of the total), partially answered 16 questions (32%), and did not obtain useful information for 14 questions (28%). Participants found answers outside the workstation to 8 of the 14 questions (57%) not answered by using the software. The most common question topic was drug information (16 questions, or 32% of the total). The most common problems encountered using the workstation were retrieval of incomplete information (20 questions, or 40% of the total) and difficulty navigating the software (16 questions, or 32%). Other problems included difficulty translating clinical problems into questions, inappropriate resource selection, inadequate training for using the software, and excessive time required to access information. The study highlights several opportunities for medical librarians and others involved in clinical information management to facilitate the use of computer software for solving clinical problems. PMID- 8428186 TI - Documenting nursing and health care history in the mid-Atlantic region. AB - The records of health care institutions can be of great value to library patrons. Yet, librarians rarely provide these unique resources because records must be collected, arranged, and described before they can be useful to patrons. The University of Pennsylvania's Center for the Study of the History of Nursing conducted a survey of health care agencies in the mid-Atlantic region to locate records created by area health care institutions. The goals of this project were to develop a database of primary source materials, to place organizational records with enduring value at suitable repositories, and to assist in the development of in-house archival programs at agencies keeping records. In-house programs provide health care institutions with a systematic way to preserve their records for administrative, legal, fiscal, and research use. Such programs also facilitate access to information, reduce cost through records management, and promote an institution through preservation and use of its historical records. The survey demonstrated that record keeping is not coordinated in most institutions, and that institutional awareness of the organization or content of records is minimal. PMID- 8428187 TI - Quality filtering of the clinical literature by librarians and physicians. AB - A study was conducted at the University of Pittsburgh to determine the effectiveness of the selection process by clinical medical librarians and to identify the criteria used by librarians and physicians to select relevant articles. The study analyzed the similarity between librarian and physician selections, the decision-making processes used by librarians and physicians, and the utility of librarian selections versus those of physicians. No significant difference in utility between librarian and physician selection was found, suggesting that librarians can recognize and select useful articles as effectively as physicians. Both librarians and physicians based selection decisions primarily on article title, abstract, and journal title. Librarians were more likely to focus on Medical Subject Headings (MeSH) descriptors, while physicians focused on clinical applicability or similarity to a specific case. Journal selection data indicate that the principle internal medicine journals were the most frequently selected sources. The study demonstrates that librarians can effectively serve a quality filtering function in the clinical environment, and they should consider extending quality filtering activities to other arenas. PMID- 8428188 TI - Citation patterns in the health sciences: implications for serials/monographic fund allocation. AB - This study sought to determine optimal serial-to-monograph ratios for collection development by comparing citation frequency with current library practice. Internal medicine literature cited an average of 88% serial references and 12% monographs. In an observational study, teaching physicians on internal medicine rounds cited 89.5% serials and 10.5% monographs to student teams. By contrast, health sciences libraries included in the Houston statistics spend an average of 79% of acquisitions budgets for serials and 21% for monographs. An 88:12 acquisitions budget ratio would be more appropriate, reflecting actual use of serials and monographs in the health sciences. PMID- 8428189 TI - Biomedical journal title changes: reasons, trends, and impact. AB - A study was conducted to document the impact of biomedical journal title changes on medical libraries and to increase awareness of the reasons titles are changed. The study consisted of two parts: a survey of academic health sciences libraries in the United States and Canada and an analysis of title changes from two different years. The survey response rate was 83%. The majority of respondents commented on difficulties in identifying and processing title changes, often resulting in the delay or loss of information. The analysis revealed that a third of title changes were not justified by the journal. The study results substantiate the need to standardize title change reporting by publishers. A standard developed by the National Information Standards Organization requires publishers to conform to standardized practices for notification. This standard precisely reflects the concerns reflected in both the survey and the study of title changes, and librarians are urged to ensure that the standard is implemented by publishers. PMID- 8428190 TI - Expanding the DOCLINE network to include nonmedical libraries in the state of Nevada. AB - Most libraries cannot meet patron demands for biomedical information using only their in-house collections. Consequently, many types of libraries request biomedical information through interlibrary loan, and these include not only academic health sciences libraries, hospital and special libraries, but also general libraries. In Nevada, with its small population spread over a large geographic area, it has become critical to develop a statewide network for sharing biomedical information. As the state resource library, the Savitt Medical Library launched an effort to establish a network, via DOCLINE, of all Nevada libraries that have health-related collections. The process of convincing academic and community college libraries to join DOCLINE and the resulting benefits of improved resource sharing and cooperative collection development are discussed. PMID- 8428191 TI - Incorrect citations: a comparison of library literature with medical literature. PMID- 8428192 TI - How hyper are we? A look at hypermedia management in academic health sciences libraries. AB - Advances in instruction-delivery technology have a direct impact on academic media centers. New technology challenges librarians philosophically, financially, and ethically to provide access to information and instructional systems. Each institution has a unique set of circumstances governing decisions to provide access to hypermedia. If patron needs are met satisfactorily through labs outside the library, it may not be necessary for the library to incorporate hypermedia into its collection. Other library media centers may serve as a main point of access, or a substantial alternative computing resource may exist in departments or professional schools. Regardless of which route is taken, hypermedia is a viable instructional delivery system and can coexist with traditional services. Future studies on various aspects of hypermedia and multimedia management should be encouraged. Academic health sciences librarians would benefit from the study of hypermedia and multimedia collection-development policies, equipment, and personnel management. As computer networking of multimedia and image databases becomes available, it will be interesting to see the role academic health sciences libraries assume in integrating these data-bases with traditional information-delivery systems. Changing technology and instructional methods will affect budgets as well as library relationships with academic departments and computing centers. PMID- 8428193 TI - End-user searching: review of a modular program. PMID- 8428194 TI - A comparison of MEDLINE CD-ROM and librarian-mediated search service users. AB - Respondents performed searches primarily for themselves and for academic research. Overall, they preferred librarian-mediated searching to CD-ROM searching. Respondents who preferred the former did so because librarians are more familiar with MeSH headings and search strategies and because of time constraints. Respondents who preferred CD-ROM liked doing their own searches and the fact that there is no cost involved. While respondents preferred librarian mediated searching over CD-ROM searching, overall they used CD-ROM more often, presumably because of other factors, such as time constraints and cost. This dichotomy could have significant implications for a library. Should the library strive to make mediated searches more attractive by providing immediate and cost free results? Or, because CD-ROM is used more, should the library continue to purchase in this area and possibly reduce support for librarian-mediated overhead (e.g., training, search tools, etc.)? And do librarians have a responsibility to encourage patrons to use the tool that offers the higher quality of retrieval? PMID- 8428196 TI - The National Library of Medicine and its role. PMID- 8428195 TI - Creating and using a faculty interests database. PMID- 8428197 TI - The GRATEFUL MED program and the medical library profession. PMID- 8428198 TI - Salaries and professional authority: new jurisdictions. PMID- 8428199 TI - Selective inhibition of agonist-induced but not shear stress-dependent release of endothelial autacoids by thapsigargin. AB - 1. The effects of the Ca(2+)-ATPase inhibitor, thapsigargin, on the shear stress dependent and on the agonist-stimulated release of endothelium-derived relaxing factor, i.e. nitric oxide (NO), and prostacyclin (PGI2) were studied in bovine and human cultured endothelial cells as well as in endothelium-intact arterial segments of the rabbit. 2. Preincubation with thapsigargin (1 microM for 10 min) had no effect on the shear stress-dependent release of NO from bovine aortic endothelial cells grown on beads, but abolished the release of NO induced by ADP, bradykinin, ionomycin or poly-L-lysine. Similarly, thapsigargin completely abrogated the agonist-stimulated PGI2 release from these cells, but had no effect on the shear stress-dependent release of PGI2. 3. The acetylcholine-induced release of NO from the luminally perfused thoracic aorta and femoral artery of the rabbit was suppressed by pretreatment with thapsigargin (1 microM). In contrast, thapsigargin did not affect the shear stress-dependent release of NO from the femoral artery. 4. Administration of thapsigargin to these vascular preparations or to cultured endothelial cells alone produced a substantial release of both NO and PGI2. This release declined towards previous values after washout of thapsigargin. 5. In human and bovine cultured endothelial cells, thapsigargin (1-1000 nM) caused a dose-dependent sustained rise in [Ca2+]i, an effect that was abolished in the absence of extracellular Ca2+. Stimulation of these cells with bradykinin, histamine, ADP or ionomycin after previous exposure to thapsigargin (30-1000 nM) no longer caused an increase in [Ca2+]i. of the release of these endothelial autacoids caused by shear stress or receptor dependent and independent agonists. PMID- 8428200 TI - Chloride anion concentration as a determinant of renal vascular responsiveness to vasoconstrictor agents. AB - 1. The role of chloride concentration in modulating vasoconstrictor responses of the rat isolated kidney, perfused with Krebs-Henseleit solution, to angiotensin II (AII), arginine vasopressin (AVP) and phenylephrine (PE) was investigated. 2. Reduction of perfusate chloride from a high (117 mM) to low (87 mM) concentration, by substitution of sodium chloride with a mixture of sodium salts of propionate, acetate and methanesulphonate, reduced responsiveness to all three vasoconstrictors, the change for AII being most pronounced. 3. For AII, reduced vasoactivity with low chloride was evident both in terms of the threshold dose and on the linear part of the dose-response curve but not for the maximum response. This attenuating effect of low chloride on the vasoconstrictor response to AII was reversed when perfusion with high chloride was reinstituted. Continuous perfusion with high chloride progressively increased the vasoconstrictor effect of low doses of AII for successive dose-response curves. 4. In addition to reducing responses on the linear part of the dose-response curve for both AVP and PE, low chloride also reduced the maximum vasoconstrictor response to PE, whereas the threshold dose for the two agonists was unchanged. In contrast to the enhanced pressor response to AII, during continuous perfusion with high chloride, tachyphylaxis occurred with AVP and PE. 5. The ability of chloride to modify renal responsiveness to vasoconstrictor agents may contribute to the increase in renal vascular resistance and decrease in glomerular filtration rate (GFR) which occurs during infusion of hyperchloremic solutions into the renal artery and explain the need for chloride as the anion accompanying sodium in salt-sensitive hypertensive models. PMID- 8428201 TI - Antiarrhythmic agents act differently on the activation phase of the ACh-response in guinea-pig atrial myocytes. AB - 1. Anti-acetylcholine effects of pilsicainide, flecainide, disopyramide and propafenone on the acetylcholine (ACh)-induced K+ current (IK.ACh) were examined in dissociated guinea-pig atrial myocytes under whole-cell voltage clamp by the use of the 'concentration-clamp' technique. 2. The IK.ACh was activated with a latency of about 100 ms after 1 microM ACh application and desensitized to a steady-state level. The latent period and the time to peak response were shortened with increasing ACh concentration. 3. The values of half-maximal inhibition (IC50) on the peak and steady state responses were 25 and 25 microM for pilsicainide, 1.7 and 2.0 microM for disopyramide, 19 and 2.0 microM for flecainide and 0.7 and 0.2 microM for propafenone, respectively. 4. Pilsicainide and disopyramide increased the latent period and the time to peak of IK.ACh in a concentration-dependent manner. Flecainide and propafenone did not change the latent period, but shortened the time to peak and hastened the decay of IK.ACh in a voltage-independent manner. 5. The results suggest that the mechanisms underlying the anti-acetylcholine effect of antiarrhythmic drugs are different among these drugs: i.e., pilsicainide and disopyramide mainly block the muscarinic ACh receptors while flecainide and propafenone inhibit the K+ channel itself as open channel blockers. PMID- 8428202 TI - Functional characterization of a 5-HT3 receptor which modulates the release of 5 HT in the guinea-pig brain. AB - 1. The aims of the present study were to confirm the modulation by 5-HT3 receptors of the electrically evoked release of tritium from slices preloaded with [3H]-5-HT of guinea-pig frontal cortex, hippocampus and hypothalamus, and to assess their functional role in 5-HT release. 2. The selective 5-HT3 agonist, 2 methyl-5-HT, introduced 8 min before the electrical stimulation, enhanced in a concentration-dependent manner the evoked release of [3H]-5-HT in the three brain regions studied. The 5-HT3 agonists, phenylbiguanide and m-chlorophenyl biguanide, did not enhance the release of tritium in frontal cortex and hypothalamus slices. 3. In hypothalamus slices, this response was lost when 2 methyl-5-HT was introduced 20 min before the stimulation, thus indicating that these 5-HT3 receptors desensitize rapidly. When 2-methyl-5-HT was added 20-min before the first stimulation period to desensitize the 5-HT3 receptors, removed for 24 min, and then re-introduced 8 min before the second stimulation period, the enhancing effect of 2-methyl-5-HT was restored, thus indicating that these 5 HT3 receptors can rapidly regain normal sensitivity. 4. The enhancing effect of 2 methyl-5-HT was attenuated by the 5-HT3 receptor antagonists m-chloro phenylpiperazine = quipazine = ondansetron > or = ICS 205-930 = BRL 24924 > MDL 72222 = zacopride. 5. The 5-HT reuptake blocker, paroxetine, enhanced the electrically evoked release of tritium when introduced 8 min before stimulation; this effect of paroxetine was blocked by ICS 205-930, thus indicating that these 5-HT3 receptors can be activated by endogenous 5-HT. 6. In the absence of electrical stimulation, 2-methyl-5-HT (10 microM) produced a marked enhancement of the basal release of [3H]-5-HT which was calcium-dependent and blocked by S zacopride but not by paroxetine. 7. The enhancing effect of 2-methyl-5-HT was dependent both on the frequency of stimulation, as indicated by the attenuated effect of 120 stimulations delivered at 1 Hz instead of 5 Hz, and on the duration of the stimulation, as indicated by the more pronounced effect of pulses delivered at 5 Hz for 24 s instead of 72 s or 120 s. PMID- 8428203 TI - Mediation by the same muscarinic receptor subtype of phasic and tonic contractile activities in the rat isolated portal vein. AB - 1. The effects of several agonists on the phasic and tonic contractile responses to muscarinic receptor stimulation have been investigated in the rat portal vein in vitro. 2. Neither chemical denervation with 6-hydroxydopamine nor the presence of the alpha 1-adrenoceptor antagonist, prazosin, influenced the spontaneous or the stimulated myogenic activity of the portal vein. 3. Indomethacin and NG-nitro L-arginine were used to investigate the influence of vasoactive factors in this preparation. They slightly increased the frequency and the amplitude of the spontaneous myogenic activity of the portal vein, respectively. NG-nitro-L arginine but not indomethacin enhanced the maximal phasic response to carbachol. Both indomethacin and NG-nitro-L-arginine failed to influence the tonic response to carbachol. 4. Muscarinic agonists increased phasic activity according to the rank order of potency: acetylcholine > muscarine > methacholine > carbachol > aceclidine > bethanechol. These effects were superimposed on a sustained contracture at higher concentrations. Oxotremorine was more potent than arecoline in increasing the mechanical phasic activity, without inducing a sustained contracture. Pilocarpine and McN A343 were weak agonists, producing submaximal effects only on phasic activity. 5. The muscarinic antagonists AF-DX116, 4 diphenylacetoxy-N-methylpiperidine (4-DAMP), P-fluorohexahydrosiladiphenidol (pFHHSiD) and pirenzepine antagonized the phasic and tonic mechanical responses to carbachol. Although the tonic contracture was slightly more sensitive to all antagonists studied, the rank order of potency: 4-DAMP > pFHHSiD > pirenzepine > AF-DX 116 was the same for both types of responses, which is indicative of a M3 receptor subtype. 6. The tonic contractile response of the rat portal vein to carbachol was more susceptible to partial receptor inactivation with propylbenzilylcholine mustard than the phasic contractile response. The dissociation constants (KA) obtained from an analysis according to Furchgott & Bursztyn (1967) were found to be 4.32 +/- 0.31 1AM for the phasic and 3.56 +/- 0.21 1AM for the tonic type of carbachol-induced response, respectively. Since the EC50-values for both carbachol-induced effects were different (phasic0.232 +/ 0.02 1AM; tonic 2.75 +/- 0.1 1AM) the phasic type of response appears to involve a large receptor reserve. PMID- 8428204 TI - Selective IK blockade as an antiarrhythmic mechanism: effects of UK66,914 on ischaemia and reperfusion arrhythmias in rat and rabbit hearts. AB - 1. UK66,914 is a specific and selective blocker of the delayed rectifying potassium current (IK). The effectiveness of IK block as a mechanism for prevention of ischaemia- and reperfusion-induced arrhythmias was tested by use of UK66,914: its actions in rat, a species deficient in cardiac IK were compared with its actions in rabbit, a species possessing functional cardiac IK. Antiarrhythmic actions in rabbit but none in rat is the only outcome possible if selective IK blockade is responsible for the antiarrhythmic actions of the drug during ischaemia and/or reperfusion. 2. During 30 min regional ischaemia, 0.3 and 1 microM UK66,914 had no influence on the incidence of ventricular fibrillation (VF) in rat (n = 9/group), values being 78% in controls, 100% in 0.3 microM treated hearts and 78% in 1.0 microM-treated hearts (NS). UK66,914 also had no effect on reperfusion-induced VF incidence (100% in each group), nor on the latency to onset of ischaemia- or reperfusion-induced arrhythmias. In contrast, in rabbit (n = 13/group), similar concentrations of drug reduced the incidence of reperfusion-induced VF from 77% in controls, to 38% and 31% (P < 0.05) respectively. The incidence of ischaemia-induced arrhythmias was too low in controls to permit detection of an antiarrhythmic effect in rabbit; however no drug-induced proarrhythmia was seen.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428205 TI - Renal selective N-acetyl-L-gamma-glutamyl prodrugs: studies on the selectivity of some model prodrugs. AB - 1. In this study, a number of structurally different N-acetyl-L-gamma-glutamyl prodrugs were investigated with respect to selective uptake by the kidney in male Wistar rats. 2. All prodrugs were tested in vitro in rat kidney slices and kidney homogenate to study their uptake and conversion. It was found that the prodrugs of para-nitroaniline (agPNA), aminophenyl acetic acid (agAFA), sulphamethoxazole (agSM), sulphadimethoxine (agSDM), propranolol (agPP) and metoprolol (agMP) were accumulated by a probenecid-sensitive carrier. The prodrug of 4'-aminoantipyrine (agAAP) was not accumulated by a probenecid- or buthionine sulphoximine-sensitive carrier. Unlike all other prodrugs, agAAP and agMP were not, or only a very limited extent converted to the parent compound in vitro. 3. agPNA, agAFA and agPP were also investigated in vivo. The tissue distribution of the prodrugs and the parent drugs was established, as was their urinary excretion and pharmacokinetic behaviour. agPNA and agAFA showed selective uptake by the kidney, in contrast to agPP which accumulated in the liver. The distribution of the parent compounds following prodrug administration was as follows: agPNA was found in kidney and plasma: agAFA in kidney only; agPP in liver only. 4. The factors which determine the selectivity of N-acetyl-L-gamma-glutamyl prodrugs are discussed. The main factors are: the transport into the kidney, the conversion rate, the residence time of the prodrug in the kidney and the presence or absence of competition for uptake and conversation by other tissues, e.g. the liver. It is concluded that this prodrug approach offers the possibility of delivering drugs selectively to the kidney, but also that it is not universally applicable. PMID- 8428206 TI - Differential sensitivity of antinociceptive assays to the bradykinin antagonist Hoe 140. AB - 1. The antinociceptive activity of the bradykinin (BK) BK2 receptor antagonist D Arg-[Hyp3,Thi5D-Tic7,Oic8]BK (Hoe 140) was determined in a range of mouse abdominal constriction assays. 2. Hoe 140 potently inhibited the response induced by i.p. injection of 10 micrograms BK/mouse, and 1 microgram BK/mouse in mice pre sensitized by i.p. injection of prostaglandin E2 (PGE2). The ED50 values in these assays were 1.9 and 3.7 micrograms kg-1 respectively. This confirms that Hoe 140 is a potent antagonist of BK in vivo. 3. Hoe 140 produced potent, but incomplete inhibition of the responses evoked by i.p. injection of kaolin or 0.25% acetic acid. ED25 values in these assays were 2.7 and 16.1 micrograms kg-1, and the maximum inhibition produced was 60% and 70% respectively. 4. At doses up to 1 mg kg-1, Hoe 140 was completely ineffective against the abdominal constriction response induced by zymosan. In contrast, morphine, ibuprofen and indomethacin had similar potencies against zymosan, kaolin and acetic acid-induced abdominal constriction. 5. Although zymosan, acetic acid and kaolin all produce qualitatively similar responses, it is appears that they achieve this by different mechanisms. The extent to which BK is involved as a mediator differs between the various types of abdominal constriction assay. PMID- 8428207 TI - Potentiation of miniature endplate potential frequency by ATP in Xenopus tadpoles. AB - 1. Extracellular application of ATP (1 mM), a substance co-stored and co-released with acetylcholine in peripheral nervous systems, potentiated the spontaneous secretion of acetylcholine (ACh) but had no effect on the amplitude and decay time constant of miniature endplate potentials (m.e.p.ps) at neuromuscular synapses in Xenopus tadpoles. 2. alpha,beta-Methylene ATP (0.3 mM) and GTP (1 mM) were also effective in increasing m.e.p.p. frequency. On the other hand, ADP, AMP and adenosine (all at 1 mM) decreased m.e.p.p. frequency. 3. Unlike the transient effect of ATP analogue and GTP on m.e.p.p. frequency, the phorbol ester TPA (2 microM) which is a protein kinase C activator, increased m.e.p.p. frequency consistently and the effects lasted as long as the presence of TPA. 4. Staurosporine (0.5 microM) and H-7 (10 microM), which are protein kinase C inhibitors, each decreased the basal level of m.e.p.p. frequency and markedly inhibited the effects of both ATP and TPA. 5. These results suggest that there is a basal activity of cytosolic protein kinases in the nerve terminals of Xenopus tadpoles and the effect of ATP is probably mediated by the binding of membrane surface purinoceptors which in turn activates cytosolic protein kinases and increases ACh release. PMID- 8428208 TI - A factor released by monocytes in the presence of dexamethasone stimulates neutrophil locomotion. AB - 1. Steroid-treated monocyte supernatants cause a dramatic increase in the speed of locomotion of human neutrophils and a significant decrease in their adhesion to protein-coated glass. In contrast, control monocyte supernatants have a smaller effect on the speed of locomotion, but cause a large increase in their adhesiveness. 2. This supernatant activity was produced equally well in the presence or absence of serum after 24 h culture at 37 degrees C with 10(-6) M dexamethasone. 3. The effect of the steroid-treated monocyte supernatants on the speed of locomotion of human peripheral blood neutrophils was not altered by rabbit polyclonal antisera against lipocortins 1-6. 4. Rabbit anti-interleukin-8 antibody which blocked the effect of IL-8 on the speed of locomotion of neutrophils did not antagonize the locomotion stimulating action of steroid treated monocyte supernatants. 5. The exocellular release of this factor(s) by human mononuclear leucocytes suggests that it may be an in vivo mediator of the anti-inflammatory effect of glucocorticoids. PMID- 8428209 TI - Effects of hyperkalaemia on the depression of maximum rate of depolarization by class I antiarrhythmic agents in guinea-pig myocardium. AB - 1 Standard microelectrode methods were used to record intracellular action potentials from strips of guinea-pig right ventricular myocardium superfused with either standard physiological saline ([K+] = 5.6 mM) or the same solution modified to contain [K+] = 11.2 mM. 2 The effects on action potential parameters of three therapeutic concentrations of mexiletine, quinidine and disopyramide were studied under both conditions at four different drive rates (interstimulus intervals = 2400, 1200, 600 and 300 ms). 3 Hyperkalaemia in the absence of drugs produced reductions in resting potential (-86.7 +/- 2.5 mV to -71.8 +/- 3.7 mV; n = 30; P < 0.001), maximum rate of depolarization (300 +/- 46.5 V s-1 to 205.6 +/- 37.6 V s-1; P < 0.0001), and action potential duration (205 +/- 26 ms to 188 +/- 32 ms; P < 0.05). 4 All three drugs produced increased depression of maximum rate of depolarization in hyperkalaemia compared to control conditions, but at all three concentrations this enhancement of effect was greater for mexiletine than for quinidine, with disopyramide exhibiting intermediate behaviour. 5 Mexiletine behaved very similarly to therapeutic concentrations of lignocaine as described in previous reports from this laboratory. 6 Quinidine behaved very similarly to Class Ic agents. 7 It is concluded that mexiletine demonstrated significantly greater selectivity for depolarized myocardium than quinidine and that this may have implications in terms of proarrhythmic potential. 8 Disopyramide exhibited intermediate selectivity for depolarized myocardium between mexiletine and quinidine. PMID- 8428210 TI - Differential effects of hydroxocobalamin on NO-mediated relaxations in rat aorta and anococcygeus muscle. AB - In rat aortic rings, hydroxocobalamin (10-30 microM) produced concentration dependent reductions of the relaxant action of nitric oxide (NO) and the endothelium-dependent, NO-mediated, relaxant action of acetylcholine. In anococcygeus muscles, hydroxocobalamin (10-30 microM) reduced but also prolonged, NO-induced relaxations, but had no effect on non-adrenergic, non-cholinergic mediated relaxations. Hydroxocobalamin had no effect on the NO-independent relaxant action of papaverine in either tissue. It is suggested that hydroxocobalamin sequesters NO by forming nitrosocobalamin. Nitrosocobalamin did not relax aortic rings, but produced a slowly developing and prolonged relaxation of anococcygeus muscles. PMID- 8428211 TI - Role of extracellular and intracellular sources of Ca2+ in sarafotoxin S6b induced contraction of strips of the rat aorta. AB - 1. The effect of sarafotoxin S6b (sarafotoxin), a vasoconstrictor peptide, on cytosolic Ca2+ concentration ([Ca2+]i) and force in rat aortic strips loaded with fura-2 was determined by front-surface fluorometry. The objective was to elucidate the role of extracellular and intracellular Ca2+ in the mechanism of action of this peptide. 2. In the presence of extracellular 1.25 mM Ca2+, sarafotoxin induced a biphasic response consisting of an initial rapid increase in [Ca2+]i followed by a secondary sustained increase. Tension developed slowly but was sustained during the application of sarafotoxin. Diltiazem (10 nM-0.1 mM) partially inhibited both the increases in [Ca2+]i and tension. 3. In the presence of extracellular Ca2+, the force developed in relation to the increase in [Ca2+]i ([Ca2+]i-force relationship) observed with sarafotoxin was much greater than that observed upon K+ depolarization. In the presence of diltiazem the sarafotoxin induced [Ca2+]i-force relationship was shifted even further to the left. 4. In the absence of extracellular Ca2+, sarafotoxin induced a transient increase in [Ca2+]i and a sustained contraction. Extending the incubation time in Ca(2+)-free physiological solution, resulted in smaller responses. However, after 60 min in Ca(2+)-free solution, sarafotoxin induced a sustained contraction but no change in [Ca2+]i. This residual contraction was inhibited by H-7, which is known to inhibit protein kinase C. 5. After treatment with caffeine to reduce intracellular stored Ca2+, sarafotoxin could still elicit increases in [Ca2+]i and in tension, showing that the caffeine-sensitive intracellular Ca2+ store partially overlaps with the sarafotoxin-sensitive store. 6. We conclude that, in addition to those components of contraction dependent on extracellular- and on intracellularly stored Ca2 , sarafotoxin can also induce contraction without increasing [Ca2+],. This component may be partially linked to the activation of protein kinase C and may contribute, in part, to the leftward shift of the [Ca2+]i-force relationship in the presence of sarafotoxin. PMID- 8428212 TI - Dependence of endotoxin-induced vascular hyporeactivity on extracellular L arginine. AB - 1. The dependence on extracellular L-arginine of vascular hyporeactivity induced by bacterial lipopolysaccharide (LPS) was studied in vivo in rats infused with LPS and in vitro in endothelium-denuded rat thoracic aortic rings exposed to LPS. 2. Infusion of LPS during 50 min at a dose of 10 mg kg-1 h-1 produced a significant impairment of the pressor effect of noradrenaline, while in tissues collected 60 min after the start of LPS infusion, no significant alteration in either plasma arginine concentration or aortic arginine content was found compared to saline-infused controls (where plasma arginine was 78.5 +/- 7 microM and aortic arginine 394 +/- 124 nmol g-1 tissue). 3. Incubation of isolated, endothelium-denuded aortic rings with LPS (10 micrograms ml-1) in the absence of L-arginine for 4 h at 37 degrees C produced a 6 fold (P < 0.01) rightward shift in the noradrenaline concentration-effect curve compared to polymyxin B (1 micrograms ml-1, a LPS neutralizing agent) and reduced by 15% the maximum observed tension. 4. The presence of L-arginine (100 microM) during the incubation with LPS and throughout the following contraction experiments caused a 15 fold (P < 0.01) increase in the EC50 of noradrenaline and greater depression (45%) of the maximum observed tension compared to polymyxin B-treated controls. Responses in control, non LPS-treated rings were unaffected by the presence of L arginine. 5. The addition of L-arginine to rings incubated with LPS in the absence of L-arginine and maximally precontracted with noradrenaline (10 microM) induced a dose-dependent relaxation. The EC50 of L-arginine was 8.0+/-0.3mu.6. The reactivity of LPS-treated rings to noradrenaline both in the absence and presence of L-arginine was restored to control levels by N0-nitro-L-arginine methyl ester (L-NAME, 300 mu), an inhibitor of NO production and by methylene blue (3 JAM), an inhibitor of guanylate cyclase.7. Incubation of isolated aortae in the absence of L-arginine did not significantly decrease the tissue arginine content, whether LPS (10 fg ml-') was present or not. Similarly, the presence of L-arginine(100 mu) in the incubation medium did not modify the tissue arginine content.8. These results show that the LPS-induced impairment of vasoconstriction elicited by noradrenaline is dependent on extracellular L-arginine, although the tissue arginine content is not depleted after LPS pretreatment, and that circulating L-arginine is sufficient to activate maximally the vascular L arginine/NO pathway in endotoxaemic rats. PMID- 8428213 TI - Comparison of the effects of selective inhibitors of phosphodiesterase types III and IV in airway smooth muscle with differing beta-adrenoceptor subtypes. AB - 1. The relaxant properties of the type IV adenosine 3',5'-cyclic monophosphate phosphodiesterase (cyclic AMP PDE) inhibitor, rolipram and the beta 2-selective and non-selective beta-adrenoceptor agonists salbutamol and isoprenaline, were compared on the guinea-pig, bovine, and mouse trachea and porcine bronchus all precontracted with methacholine (EC30). 2. Rolipram and both beta-agonists produced concentration-dependent reversal of the methacholine-induced tone in the four airway preparations. 3. Isoprenaline and salbutamol were similar in potency on the guinea-pig (-log10IC50:8.43, 8.06) and bovine (-log10 IC50:8.52, 8.40) airways. In contrast, salbutamol was much less potent than isoprenaline on the mouse trachea (> 1000 fold) and the porcine bronchus (> 100,000 fold). 4. The potency of rolipram approached that of isoprenaline on the guinea-pig and bovine trachea (beta 2-adrenoceptors predominate). However, rolipram was significantly less active than isoprenaline on the porcine bronchus (1000 fold) and mouse trachea (> 2000 fold) where beta 2-adrenoceptors predominate. 5. Siguazodan, the type III cyclic AMP PDE inhibitor, produced concentration-dependent relaxations of the porcine bronchus and guinea-pig trachea contracted with methacholine. Siguazodan was 100 fold more active than rolipram in pig tissues indicating the type III isoenzyme may be of greater functional significance in this tissue. In contrast, siguazodan was 15 times less potent that rolipram in guinea-pig airways suggesting a greater role for the type IV PDE. 6. These findings may reflect a possible relationship between the beta 2-adrenoceptor subtype and the functional importance of the type IV PDE isoenzyme. A similar relationship may exist between beta 1-adrenoceptors and the PDE type III isoenzyme. PMID- 8428214 TI - Protein kinase C inhibitors enhance endothelin-1 and attenuate vasopressin and angiotensin II evoked [Ca2+]i elevation in the rat cardiomyocyte. AB - Primary cultures of neonatal rat cardiomyocytes were pretreated for 16 h with either nonselective (staurosporine, 100 nM) or selective (NPC15437, 20 microM) protein kinase C (PKC) inhibitors. These inhibitors did not affect the basal cytosolic free calcium, [Ca2+]i, level (106 +/- 12 nM) as determined by fura-2 fluorescence methodology. Both agents significantly enhanced the maximal [Ca2+]i responses to endothelin-1 (ET-1) and attenuated the peak [Ca2+]i responses to arginine vasopressin and angiotensin II. They did not alter the EC50 values of any of these agonists. Since depletion of [Ca2+]o led to only partial attenuation of the enhanced response to ET-1 in the treatment groups, it is likely that PKC inhibition results in an exaggerated intracellular mobilization of Ca2+ to ET-1. It is concluded that PKC modulates agonist(s)-evoked intracellular Ca2+ mobilization and that the nature of regulation is governed by the agonist. PMID- 8428215 TI - The local anti-inflammatory action of dexamethasone in the rat carrageenin oedema model is reversed by an antiserum to lipocortin 1. AB - 1. A local pre-injection of 1 micrograms dexamethasone sodium phosphate strongly inhibited (> 60% inhibition at 3 h; P < 0.001 at all time points) the development of carrageenin-induced paw oedema in the rat induced by a subplantar injection of 0.1 ml, 2% carrageenin. 2. Coinjection of a polyclonal rabbit antiserum raised against human 1-188 recombinant lipocortin 1, which also recognised the rat protein, reversed the inhibitory action of dexamethasone (P < 0.05 at 4 h and 5 h). At the highest volume used (40 microliters) control antisera were without any effect. 3. These data further support the concept that lipocortin 1 is involved in the anti-inflammatory mechanism of action of the glucocorticoids. PMID- 8428216 TI - Glucocorticoid-and non-glucocorticoid induction of lipocortins (annexins) 1 and 2 in rat peritoneal leucocytes in vivo. AB - 1. We have studied the occurrence, distribution and disposition of lipocortins (annexins) 1, 2 and 5 in mixed peritoneal leucocytes obtained from rats in which glucocorticoid levels were altered by adrenalectomy, administration of the glucocorticoid antagonist, RU486, or by injection of dexamethasone or hydrocortisone, as well as from rats in which the peritoneal cells were elicited by inflammatory stimuli. 2. In cells obtained from untreated rats with an intact adrenal cortex, lipocortins 1, 2 and 5 were readily detectable: the majority of each of the proteins was apparently located intracellularly with much smaller amounts in the membrane. Lipocortin 1 and to a lesser extent lipocortin 5 were also seen in a Ca(2+)-dependent association with the external plasma membrane. Following administration of RU486 (2 x 20 mg kg-1) the amounts of lipocortin 1 and 2 in cells were greatly reduced. Conversely, injection of hydrocortisone (1 mg kg-1) or dexamethasone (0.08 mg kg-1) caused an increase in the amount of lipocortin 1 and 2 in peritoneal cells within 30 min. Lipocortin 5 was unchanged by any manipulation of glucocorticoid levels. 3. Lipocortins 1 and 2 were elevated in both intracellular and membrane-associated fractions of macrophages elicited by intraperitoneal injection in inflammogens. This phenomenon also occurred in adrenalectomized animals. 4. Our data indicate that glucocorticoids control the synthesis of some members of the lipocortin family in rat mixed peritoneal cells but also suggest the existence of a separate system for controlling the generation of this protein. The significance of these observations is considered in relation to the mechanism of glucocorticoid hormone action on eicosanoid production. PMID- 8428217 TI - The actions of nitric oxide donors in the prevention or induction of injury to the rat gastric mucosa. AB - 1. The protective or damaging actions on the gastric mucosa, of locally infused nitrovasodilators that donate nitric oxide (NO), have been investigated in the pentobarbitone-anaesthetized rat. 2. Local intra-arterial infusion of endothelin 1 (ET-1; 5 pmol kg-1 min-1 for 10 min) induced extensive, macroscopically apparent, haemorrhagic injury to the rat gastric mucosa. This damage was dose dependently reduced by concurrent local intra-arterial infusion of glyceryl trinitrate (GTN; 10-40 micrograms kg-1 min-1) which liberates NO on metabolic transformation, or the nitrosothiol, S-nitroso-N-acetyl-penicillamine (SNAP, 2.5 10 micrograms kg-1 min-1) which spontaneously liberates NO. 3. Local infusion of higher doses of SNAP (20 and 40 micrograms kg-1 min-1, i.a.) did not, however, significantly protect against mucosal injury induced by ET-1. 4. Furthermore, local infusion alone of these higher doses of SNAP, as well as sodium nitroprusside (10-40 micrograms kg-1 min-1, i.a.) which also spontaneously liberates NO, induced significant mucosal injury, as assessed macroscopically and confirmed by histology. 5. Local infusion of these higher doses of SNAP and nitroprusside reduced systemic arterial blood pressure (BP), but this was not correlated with the extent of mucosal injury. 6. Furthermore, local infusion of GTN (10-40 micrograms kg-1 min-1, i.a.) alone, which also reduced BP, failed to induce gastric mucosal damage. 7. These findings suggest that exogenous NO can protect the rat gastric mucosa from damage induced by the vasoconstrictor peptide ET-1, which may reflect local microcirculatory interactions. However, the unregulated release of high levels of NO within the microvasculature induces mucosal injury. PMID- 8428218 TI - Adenosine A1-receptor stimulated increases in intracellular calcium in the smooth muscle cell line, DDT1MF-2. AB - 1. The effect of a range of adenosine receptor agonists on intracellular free calcium concentration ([Ca2+]i) has been studied in the hamster vas deferens smooth muscle cell line DDT1MF-2. 2. Adenosine receptor agonists elicited a rapid and maintained increase in [Ca2+]i in fura-2 loaded DDT1MF-2 cells. The initial rise could be maintained in the absence of extracellular calcium, whereas the maintained or plateau phase was dependent upon the presence of extracellular calcium and appeared to be associated with calcium influx. The rank order of agonist potencies was N6-cyclopentyladenosine > 5'-N-ethylcarboxamidoadenosine > 2-chloroadenosine > adenosine. 3. The response to 2-chloroadenosine was antagonized by the antagonists 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, KD 0.14 nM) and 8-phenyltheophylline (KD 112 nM). 4. Pretreatment with the 5-lipoxygenase inhibitor AA861 (20 microM) produced only a small (14 +/- 2%) inhibition of the [Ca2+]i response elicted by N6-cyclopentyladenosine (300 nM), in nominally Ca(2+) free buffer containing 0.1 mM EGTA. The cyclo-oxygenase inhibitor, indomethacin (2 microM) was without effect. 5. The Ca(2+)-influx associated with the plateau phase required the continued presence of agonist on the receptor. The antagonist DPCPX (100 nM) attenuated the rise in [Ca2+]i observed when extracellular Ca2+ was re-applied after the cells had been stimulated with N6-cyclopentyladenosine (CPA; 300 nM) in experiments initiated in nominally Ca(2+)-free buffer. 6. Pretreatment with pertussis toxin (200 ng ml-1 for 4 h) inhibited the CPA (100 nM) stimulated intracellular Ca2+ release and Ca2+ influx but was without effect on the response to histamine (100 microM). 7.These data suggest that adenosine A(1)-receptor activation in DDT(1)MF-2 cells stimulates release of Ca(2+) from intracellular stores and influx of extracellular Ca(2+) through Ca(2+) entry pathways in the plasma membrane which required the continued presence of agonist on the receptor. PMID- 8428219 TI - Nitric oxide mediates the inhibition by interleukin-1 beta of pentagastrin stimulated rat gastric acid secretion. AB - Bolus injection of interleukin-1 beta (2 micrograms kg-1, i.v.) inhibited acid secretion induced by intravenous infusion of pentagastrin (8 micrograms kg-1 h-1) in the continuously perfused stomach of the anaesthetized rat. Administration of interleukin-1 beta did not modify mean systemic arterial blood pressure. Pretreatment with NG-nitro-L-arginine methyl ester (L-NAME, 2-10 mg kg-1, i.v.), but not dexamethasone (5 mg kg-1, s.c. twice over 16 h), restored the acid secretory responses to pentagastrin. The actions of L-NAME were reversed by the prior administration of L-arginine (100 mg kg-1, i.v.), but not by its enantiomer D-arginine (100 mg kg-1, i.v.). L-NAME (5 mg kg-1, i.v.) increased blood pressure but this was not the mechanism by which interleukin-induced acid inhibition was prevented, since similar systemic pressor responses induced by phenylephrine (10 micrograms kg-1 min-1, i.v.), had no such effect. These findings suggest that interleukin-induced inhibition of acid responses to pentagastrin involves synthesis of NO from L-arginine. PMID- 8428220 TI - Rheumatoid arthritis in a population of persons aged 85 years and over. AB - A Dutch urban population of 977 persons aged 85 years and over was examined for the presence of rheumatoid arthritis (RA). Prevalence rates for definite RA, past polyarthritis with joint deformation and past polyarthritis without joint deformation were 0.3%, 0.3% and 0.7%, respectively. The polyarthritis patients did not differ from age-matched controls with respect to crude scores of physical disability, the presence of serum rheumatoid factor or the presence of HLA-DR4. Although these data should be interpreted with caution because of the small patient groups, we conclude that in persons aged 85 years and over the prevalence of RA is low, and the disease is relatively mild. PMID- 8428221 TI - The prevalence and incidence of systemic lupus erythematosus in Nottingham, UK, 1989-1990. AB - The incidence and prevalence of systemic lupus erythematosus (SLE) in a well defined area in the Midlands was determined, by case ascertainment using multiple sources, during the period 1.5.89 to 30.4.90. This first such study of SLE in the UK showed incidence rates of 1.5/100,000/year for males and 6.5/100,000/year for females. The highest incidence was seen in age groups 40-49 and 50-59 years, with rates for females of 10.5 and 18.4/100,000/year respectively. Prevalence rates were 3.7/100,000/year for males and 45.4/100,000/year for females: SLE was found to be more prevalent amongst Afro-Caribbean groups. The socioeconomic status of the SLE patients was similar to the local study population, using social class by occupation and disadvantage by geographical area as indicators. Marked overlap between different sources of retrieval suggests that ascertainment of cases was high. PMID- 8428222 TI - Hypermobility: prevalence and features in a Swedish population. AB - In this investigation, parallel to an earlier study [1] of 660 musicians from the USA, 606 individuals from a factory in Ostersund, Sweden, were studied to determine the prevalence and nature of their hypermobility. They were examined using the same protocol and by the same examiner as in the earlier study for the presence of five well recognized features of hypermobility. In this mostly middle aged population of workers, as in the earlier population of mostly young musicians, joint laxity was found to be far more common among females. In both sexes hypermobility is seen to decline with age, the rate of fall being about the same for men and women. PMID- 8428223 TI - Joint hyperlaxity and musculoligamentous lesions: study of a population of homogeneous age, sex and physical exertion. AB - A total of 675 male soldiers of the same age (17 years) and on the same physical exertion regimen were studied in order to determine the incidence of joint laxity and its potential relationship to the appearance of musculoligamentous lesions. They were investigated during their 2-month military training period, which involved the same heavy physical exertion for all the individuals. The degree of joint laxity was determined on the basis of five criteria involving the hand, elbow, knee and spine. The overall population was divided into three groups according to the number of criteria met, namely: (1) normal or non-lax individuals, with none or only one criterion (67% of the studied population); (2) lax individuals, with two or three criteria (25.5%); and (3) hyperlax individuals, with four or five criteria (7.5%). The occurrence of musculoligamentous lesions during the 2-month study, particularly those involving the ankle and knee, was significantly more frequent in hyperlax and lax individuals than it was in their counterparts with normal joint mobility. These results confirm that joint hyperlaxity predisposes individuals to musculoligamentous lesions. PMID- 8428224 TI - Stimulation of von Willebrand factor antigen release by immunoglobulin from thrombosis prone patients with systemic lupus erythematosus and the anti phospholipid syndrome. AB - The level of von Willebrand factor antigen (vWF) released by endothelial cells in response to IgG isolated from 18 patients with systemic lupus erythematosus (SLE), eight patients with the anti-phospholipid syndrome (APS) and 22 controls has been measured. Incubation with IgG from the combined patient group resulted in a significantly greater release of vWF (mean stimulation index +/- SEM, 4.57 +/- 0.78) when compared with controls (1.96 +/- 0.22, P = 0.003). Furthermore, IgG from 17 patients who had had a history of thrombosis induced higher levels of vWF release (5.33 +/- 1.09) when compared with the controls (P = 0.008). These findings suggest that IgG from patients with SLE or APS is capable of stimulating vWF release and that this ability may be implicated in the thrombotic events that are observed in these conditions. PMID- 8428225 TI - The influence of arthrogenous muscle inhibition on quadriceps rehabilitation of patients with early, unilateral osteoarthritic knees. AB - Reflex arthrogenous muscle inhibition (AMI) may cause muscle atrophy or impede effective rehabilitation of affected muscle groups. To investigate this, bilateral quadriceps AMI, isometric and isokinetic muscle strength were measured in 10 patients with unilateral osteoarthritic knees, before and after a course of routine physiotherapy. Before rehabilitation, quadriceps of all the diseased legs were inhibited (P < 0.05) and 40% weaker (P < 0.02) than the non-diseased legs. Following rehabilitation, AMI decreased (P < 0.01) in the diseased leg and strength increased at all test velocities (P < 0.05-0.005); however, strength deficits compared with the non-diseased leg remained. Subjective improvements in functional ability and confidence in the diseased leg were reported. Though AMI may be partially responsible for unilateral muscle weakness, it does not preclude strength gain in affected muscles. Possible physiological mechanisms which evoke AMI may also adversely affect muscle proprioception, implicating AMI as a possible cause of initiation or progression of degenerative joint disease. PMID- 8428226 TI - The Health Assessment Questionnaire as a predictor of non-steroidal peptic ulceration. AB - To assess the possibility of a relationship between the degree of physical disability in arthritic patients and non-steroidal peptic ulceration, patients were endoscoped immediately after performing their arthritic functional assessments. The Health Assessment Questionnaire was used and patients were classified into three main groups: I, scoring 0-1; II, 1.1-2; and III, 2.1-3. Ulcers, found in 36 out of 89 patients studied (36/89, 40%), were distributed as follows: 15/22 (68%) in group III compared with 9/28 (32%) in group I (chi 2 = 5.2, P < 0.02), and 12/39 (31%) in group II (chi 2 = 7.24, P < 0.01). Patients with debilitating arthritis appear to have a higher prevalence of non-steroidal peptic ulceration. This finding might be relevant to the process of selecting patients for prophylactic anti-ulcer therapy. PMID- 8428227 TI - Primary fibromyalgia syndrome--an outcome study. AB - Seventy-two patients (65 F, 7 M; mean age 52, range 18-81 years) fulfilling criteria for primary fibromyalgia syndrome (PFS) were reviewed at a mean of 4 years (range 1.5-6) following diagnosis. Ninety-seven per cent still had symptoms typical of PFS (60% worse, 26% better than at presentation); 85% had multiple hyperalgesic tender sites and still fulfilled criteria for PFS; 8% had more limited, and 7% had no tender sites at review. Many had significant disability (median score 1.0, range 0-2.75, on the Health Assessment Questionnaire; median 2, range 1-3 on the Steinbrocker index) and 92% scored highly (> 12) on the Hospital Anxiety and Depression index. In no patient did screening investigations reveal development of inflammatory, metabolic, endocrine or muscle disease. This study confirms a poor outcome for PFS patients and association with often marked functional disability and high levels of anxiety and depression. Contrary to one previous study there was no evidence that PFS predates the onset of other disease. PMID- 8428228 TI - Two cases of 'Wegener's tuberculosis'. PMID- 8428229 TI - Wegener's granulomatosis and ANCA-associated diseases in 1992. PMID- 8428230 TI - Systemic lupus erythematosus evolving into systemic vasculitis: a report of five cases. AB - Five patients with systemic lupus erythematosus are described who developed the disease in their second or third decade. The pattern of their disease then changed markedly and they developed the clinical features of a systemic vasculitis. The possible explanations for this transition of one connective tissue disease to another is discussed. PMID- 8428231 TI - Exacerbation of systemic lupus erythematosus during human parvovirus B19 infection. PMID- 8428232 TI - Systemic sclerosis occurring in a patient with Adamantiades-Behcet's disease. AB - We report a patient who developed SS while being treated with azathioprine for ABD. This is a hitherto unreported association between the two conditions. PMID- 8428233 TI - Pigmented villonodular synovitis of the knee in a patient with Paget's disease of the adjacent femoral bone. AB - A 49-year-old male patient, diagnosed as having Paget's disease involving the lumbar vertebrae and both femora, felt persistent pain and swelling in the left knee after trauma. A synovectomy was performed. Clinical and histological findings led to the diagnosis of pigmented villonodular synovitis of the knee joint. Only one previous case of this association has been reported. A common aetiology for both diseases is discussed, and traumatic factors should be considered in the aetiology of the association. PMID- 8428234 TI - Pancreatic function in patients with primary Sjogren's syndrome. PMID- 8428235 TI - Reliability of skin score in scleroderma. PMID- 8428236 TI - Genetic factors influencing the outcome of early arthritis--the role of sulphoxidation status. PMID- 8428237 TI - Non-fatal bronchiolitis obliterans possibly associated with tiopronin. A case report with long-term follow-up. PMID- 8428238 TI - Central nervous system inflammation in the eosinophilia-myalgia syndrome. PMID- 8428239 TI - Whiplash injuries. PMID- 8428240 TI - Immunotherapy, past, present and future. PMID- 8428241 TI - Joint hyperlaxity: is there a case for screening? PMID- 8428242 TI - Unusual DQA-DR haplotypes in rheumatoid vasculitis. AB - DQA and DQB variants and HLA haplotypes were defined in Caucasian subjects with rheumatoid arthritis (RA), in a rheumatoid subset characterized clinically by extra-articular features of major vasculitis, and in controls. DQ variants were defined using a panel of sequence specific oligonucleotide probes. In RA subjects without extra-articular features the frequency of DQB*0301 was significantly increased but this was secondary to the main association with DR4. In rheumatoid vasculitis by contrast, DQB*0302 rather than DQB*0301 was increased in frequency, in addition to an increase of C4A null alleles. Family studies showed that DR4 negative haplotypes had an increased frequency of unusual DR-DQA combinations as compared to other rheumatoid subsets. These findings are in keeping with the concept that genes within the MHC other than DR4 have a disease-modifying role in rheumatoid subsets. HLA haplotypes were defined in a family where the proband has RA and Felty's syndrome and an affected sister has rheumatoid vasculitis. These siblings share a DR4 bearing haplotype typing for DQB*0301, and the sister with rheumatoid vasculitis has a DR4 negative haplotype carrying a C4A null allele and an unusual DR-DQA combination. PMID- 8428243 TI - Differentiating persistent from self-limiting symmetrical synovitis in an early arthritis clinic. AB - Early rheumatoid arthritis (RA) must be differentiated from benign self-limiting polyarthritis because of the risks associated with treatment of RA. Conventional, widely available clinical and laboratory variables, measured at first clinic visit, were studied for their ability to predict persistence in 112 patients with up to 6 months of joint symptoms. Those 65 patients with symmetrical peripheral polyarthritis were followed for 1 year: 36 who underwent spontaneous remission were classified self-limiting synovitis (SLS); the remaining 29 were termed persistent synovitis (PS). Univariate analysis suggested more severe disease in PS at presentation but showing considerable overlap with SLS, making clinical discrimination difficult. Multivariate analysis confirmed this overlap but identified a subset of most helpful variables. The RA latex was the most powerful variable, yet accounted for only 45% of the variability in outcome. Combining a positive RA latex with an ESR > 30 mm/h carried a relative risk for PS of 4.33, with specificity 94% but sensitivity only 69%. Self-limiting synovitis initially could not be distinguished from early RA: hence RA may exist in two forms, the traditional persistent form and a less well recognized abortive form. PMID- 8428244 TI - The gene as theme in the paradigm of cancer. PMID- 8428245 TI - Bowel preparation before intravenous urography: is it necessary? AB - Whether or not bowel preparation should be used before intravenous urography (IVU) remains a controversial issue. Despite strongly held views on both sides there is little scientific evidence to support either viewpoint. We have conducted a prospective randomized study designed to test the hypothesis that adequate bowel preparation before IVU facilitates better quality studies requiring fewer films and consequently less time and a lower radiation exposure. Data on 188 patients were analysed; 90 patients received bowel preparation and 98 received no bowel preparation. There was no difference between the groups in terms of the number of films taken, the duration of the procedure, the visibility of the renal tracts or the overall quality of the studies. The prepared group did have significantly less faecal residue than the unprepared group. However, the renal tract visibility was no greater, as the combination of gas and haustral folds seen after bowel preparation obscured fine detail of the urinary tract as effectively as faecal residue. The hypothesis that adequate bowel preparation before IVU facilitates better quality studies must therefore be rejected. PMID- 8428246 TI - Abnormalities in rCBF and computed tomography in patients with Alzheimer's disease and in controls. AB - The pattern of abnormal distribution of the single photon emission computed tomography (SPECT) cerebral blood flow tracer 99m-technetium hexamethylpropyleneamine oxime (99Tcm-HMPAO) was investigated in 14 patients with clinically diagnosed Alzheimer's disease (AD) who subsequently had post-mortem confirmation of the disease and also in 14 elderly control subjects. These abnormalities were compared with computed tomography (CT) scans to investigate the degree to which the focal SPECT deficits were due to atrophy. The results show that SPECT imaging with 99Tcm-HMPAO and CT scanning both have a higher incidence of abnormality in AD patients than in controls and that the difference between patients and controls is greater with SPECT than with CT. Frontal SPECT and CT abnormalities in moderate/severe Alzheimer's disease occur as frequently as temporal/occipital abnormalities but the latter are rare in control subjects. Around 50% of the SPECT deficits occur in CT normal brain regions, showing that atrophy is not the sole cause of SPECT deficits. PMID- 8428247 TI - Lymphography--still useful in the diagnosis of lymphangiomatosis. AB - Lymphangiomatosis is diagnosed by lymphography in two patients. One case presented with life threatening pleural effusion and was found to have skin haemangiomata, underlying lymphangiomatosis and massive osteolysis of the pelvic bones in the same topographic distribution. The aetiology of this condition is discussed and the role of lymphography emphasized. PMID- 8428248 TI - Imaging of dialysis access: a review of 67 failing fistulas investigated by intravenous digital subtraction angiography. AB - We have reviewed 67 failing haemodialysis fistulas imaged by intravenous digital subtraction angiography (IVDSA). Peripheral venous injection of non-ionic contrast was used in all examinations. No complications relating to peripheral injection, contrast dose or fluid load were experienced. Image quality was poor, owing to inadequate vascular opacification, in 1/67. Image quality was excellent (53/67), or good (13/67) in the remaining 66. 56/66 of these examinations were diagnostic. Diagnoses included abnormalities of feeding arteries, anastomoses, draining veins, and central veins. Multiple abnormalities were demonstrated in 10/56 examinations. Conventional arm venography was performed in addition to IVDSA in five cases where venous images were inadequate owing to anastomotic obstruction (3/5) or poor opacification (2/5). 5/66 examinations were non diagnostic owing to failure to image the central veins where no cause for fistula failure had been demonstrated. A further 5/66 examinations were non-diagnostic owing to failure to obtain oblique projections of the shunt where vascular details was obscured by overlapping vessels. The authors recommended routine imaging of the dialysis shunt in two planes and central venous imaging in all cases. This would have resulted in reduction of the failure rate from 16% to 1.5% in this series. IVDSA using a peripheral injection technique is simple, free from complications, and provides good quality images of both arterial and venous components of the fistula. Images of vessels not directly involved in fistula formation are routinely obtained and aid the planning of fistula revision. PMID- 8428249 TI - Detection of significant abnormalities on lumbar spine radiographs. AB - The purpose of this study was to determine the proportion of significant abnormalities detected on anteroposterior (AP) and lateral radiographs of the lumbar spine when viewed separately, in a series of cases where the prevalence of abnormalities had been artificially increased. Five radiologists of varying experience were required to report separately on the AP and lateral films of 300 cases in which randomly included were 30 cases with metastatic disease, a disc infection or an inflammatory spondylitis. At a later date, unaware of their initial observations, the radiologists repeated the exercise reviewing all the films together. As might be expected the false positive rate was relatively high, particularly in the least experienced radiologists' responses. Nevertheless the overall results indicate that the majority of early inflammatory spondylitis cases will be missed on a solitary lateral film as will many of the metastases. Conversely, fractionally more of the disc infections were observed on the lateral film than on the AP. Further analyses in terms of sensitivity, specificity, positive and negative predictive values are presented. The authors conclude that it is preferable to reduce the overall number of lumbar spine examinations performed, by adherence to accepted guidelines, than to prejudice the detection of significant, albeit rare, abnormalities by restricting the routine series of radiographs. PMID- 8428251 TI - Calibration of ion chambers for use in mammography. AB - There is at present no UK calibration service for ion chambers for mammography, where X-ray beams are produced from tubes having molybdenum targets and filters. This paper reports calibrations against a radiotherapy secondary standard (calibrated for beams from tungsten targets with aluminium filters) using beams from both types of target and filter. Two examples of the Radcal mammography dosimeter were found to have calibration factors which varied by less than 1% in molybdenum target beams from 30 to 40 kV. Differences between calibrations using the two types of X-ray beam did not exceed about 2%. All calibration factors were within about +/- 2% of 1.0. Errors are thought to be within +/- 3%. The results of an independent calibration of one of these dosimeters against a similar chamber calibrated by CEC are also reported. Calibrations of this kind can only be temporary expedients until adequate calibration facilities for mammography beams become available, but are nevertheless useful. PMID- 8428250 TI - Measurement of focal spot size in mammography X-ray tubes. AB - Three different measurement techniques for estimating the size of focal spots in X-ray tubes are described. These are the pin-hole, the slit and the star pattern or resolution pattern, and all are well known. Results are reported for a number of modern mammography X-ray tubes employed in a screening programme, using all three techniques. The results are compared with each other and with makers' specifications, IEC tolerances and UK Department of Health guidelines. Agreement between slit and star pattern results is generally within a few per cent, while pin-hole results are usually appreciably smaller. Although the slit technique is the most reliable for estimating focal spot size, both the others have a useful role, especially in revealing focal spot condition. Compliance with makers' specifications is usually demonstrated, but compliance with Department of Health guidelines is only found on the width of broad foci. PMID- 8428252 TI - Measurement of the bone mineral density of the os calcis as an indication of vertebral fracture in women with lumbar osteoarthritis. AB - In women with lumbar osteoarthritis, measurement of the os calcis bone mineral density (BMD) using dual-energy X-ray absorptiometry (DEXA) as an indication of vertebral fracture was evaluated. The in vivo precision of the method was 1.28%. Age- and sex-matched control curves were evaluated using a control of 193 females. The correlation between spine BMD and os calcis BMD was significant (r = 0.65, p << 0.001). For the osteoporotic women without osteoarthritis (n = 34), there was no significant difference in the spine and the os calcis Z-scores ( 1.99SD and -1.83SD respectively). Whereas for osteoporotic women with osteoarthritis (n = 30) the spine Z-score was -0.49SD the os calcis Z-score was 1.92SD. The difference was significant (p < 0.001). Receiver operating characteristic (ROC) curves demonstrate the superiority of the os calcis as a measurement site over the lumbar spine, in correlation with existing crush fractures in the presence of osteoarthritis. It is concluded that when lumbar osteoarthritis occurs measurement of the os calcis BMD using DEXA is clinically useful for the estimation of bone mass. PMID- 8428253 TI - Kinetic considerations in the choice of treatment schedules for neuraxis radiotherapy. AB - Neuraxis radiotherapy of radiosensitive tumours such as medulloblastoma is usually carried out using conventionally sized fractions and a shrinking field technique. Plowman and Doughty (Br. J. Radiol., 64 (1991) 603-607) have proposed a partial transmission block (PTB) technique which entails the use of small daily doses over a conventional time period. Radiobiological analysis suggests that, although the PTB technique may be adequate for slowly growing tumours, therapeutic efficacy is likely to be compromised where the tumour doubling time is short. Accelerated hyperfractionation (twice daily fractions) provides a possible alternative to both conventional scheduling and the PTB technique. Direct measurement of the kinetics of tumour cells in CSF, where possible, may provide useful guidance in the choice of regimes. PMID- 8428254 TI - Use of peritoneal insufflation to displace the small bowel during pelvic and abdominal radiotherapy in carcinoma of the cervix. AB - Peritoneal insufflation is a technique which reliably displaces small bowel from pelvic and abdominal radiotherapy fields with the aim of reducing bowel reactions which limit the dose of radiation applied to these sites. Use of this technique in five patients undergoing radiotherapy for advanced carcinoma of the cervix, and the degree of bowel displacement resulting, dosimetry, acute reactions and tolerability of the technique are presented, with discussion of the possibility of future escalation in radiotherapy dose. PMID- 8428255 TI - Uptake of technetium-99m methylene diphosphonate by a pulmonary hamartoma/mesenchymoma. PMID- 8428256 TI - Sclerotic bone lesions in Gorlin's syndrome. PMID- 8428257 TI - Agenesis of the corpus callosum. PMID- 8428258 TI - Ultrasound diagnosis of duodenal atresia combined with isolated oesophageal atresia. PMID- 8428259 TI - The case of the missing calcified disc. PMID- 8428260 TI - Output factors for blocked fields. PMID- 8428261 TI - Partial transmission block radiotherapy technique for childhood medulloblastoma. PMID- 8428262 TI - Role of the neutrophil in adult respiratory distress syndrome. AB - Adult respiratory distress syndrome (ARDS) remains a significant cause of morbidity and mortality in surgical practice. Despite the continued advance of surgical technique and therapy, the mainstay of treatment of ARDS remains supportive. In the past decade cytokines have been found to be primary chemical mediators of the host response to inflammatory disease. The polymorphonuclear leucocyte has also emerged as a possible cellular mediator of the end-organ damage that characterizes these inflammatory processes. The role of the neutrophil as the primary cellular mediator of alveolar capillary membrane injury in ARDS remains controversial. This article reviews the relevant current literature and considers the implications of the prevailing evidence on future management of this syndrome. PMID- 8428263 TI - Long-term follow-up after open cholecystectomy. AB - The long-term outcome of open cholecystectomy was investigated. Follow-up data at a median of 10 years were obtained from physicians' records on 325 (92.6 per cent) of 351 patients treated for symptomatic gallbladder stones by open cholecystectomy between 1978 and 1980. One 79-year-old patient died after operation, from a pulmonary complication. Of the 325 patients, 81.5 per cent were currently asymptomatic or had not had symptoms during follow-up until death. Sixty patients (18.5 per cent) had complaints after 10 years. Stone recurrence was found in five patients (1.5 per cent); ten others (3.1 per cent) had biliary tract-related complaints during follow-up. Blood samples were obtained in 67.3 per cent of surviving patients with biliary complaints. There were no laboratory findings associated with biliary obstruction. Patients with typical symptoms of gallstone disease before surgery had significantly fewer complaints during follow up than those with typical as well as atypical symptoms (14.8 versus 26.5 per cent, P < 0.02). However, most of these complaints were not related to the procedure. It is concluded that the majority of patients reported no complaints or postcholecystectomy symptoms 10 years after surgery. PMID- 8428264 TI - Minimally invasive treatment for common bile duct stones. AB - The impact of endoscopic surgery on the management of stones in the common bile duct (CBD) was studied. All patients with proven common duct stones managed by a single consultant during a 21-month period were included. Of a total of 60 patients, 48 were referred because of symptoms or signs of calculi in the CBD and 12 were found to have common duct stones although symptoms were related primarily to the gallbladder. Of the 48 patients referred with common duct stones, 41 (85 per cent) were successfully treated by endoscopic retrograde cholangiography (ERC) and endoscopic sphincterotomy (ES) (complications were cholangitis (one patient), mild acute pancreatitis (one), impacted retrieval basket (one)). Of the remaining seven patients, two underwent open exploration (one after failed laparoscopic exploration) and five had successful laparoscopic duct exploration. One patient underwent laparoscopic exploration of the CBD but subsequently died. Of the patients with primary gallbladder symptoms, eight underwent preoperative ES, one was treated by laparoscopic duct exploration, one by conversion to open operation and in two patients duct stones were removed at postoperative ERC. The overall mortality rate was 2 per cent and the incidence of complications 12 per cent. Open exploration of the CBD was performed in three patients (5 per cent). This experience suggests that open duct exploration will become increasingly infrequent and may be replaced by endoscopic surgery in the majority of patients. PMID- 8428265 TI - Plasminogen activator inhibitor 2 reduces peritoneal fibrinolytic activity in inflammation. AB - Fibrinolysis in peritoneal tissue may play a role in the development of intra abdominal adhesions. The plasminogen-activating capacity of human peritoneum results largely from the presence of tissue plasminogen activator (tPA). Inflammation reduces peritoneal plasminogen-activating activity and leads to the appearance of plasminogen activator inhibitor (PAI) type 1. The role of PAI-2 in the inhibition of peritoneal fibrinolysis during inflammation was investigated in this study. The plasminogen-activating activity of peritoneal biopsy homogenates (seven inflamed, seven normal), measured using a fibrin plate technique, was reduced in inflamed compared with normal tissue (median < 0.07 versus 13.9 units/cm2, P < 0.01); tPA antigen levels were not significantly different (median 1.02 versus 1.34 ng/ml). PAI-1 and PAI-2 antigens were not detected in normal human peritoneum but were present in inflamed peritoneum (median concentration 8.8 ng/ml for PAI-1, 26.7 ng/ml for PAI-2). These inhibitors may be important factors in adhesion formation by contributing to the abolition of peritoneal plasminogen-activating activity. PMID- 8428266 TI - Laparoscopically assisted reversal of Hartmann's procedure. PMID- 8428267 TI - Controlled trial of wound infiltration with bupivacaine for postoperative pain relief after appendicectomy in children. AB - A double-blind controlled trial was performed to assess the effectiveness of wound infiltration with bupivacaine for postoperative pain relief after appendicectomy in children. Sixty children entered the trial and 52 completed it; they were randomized to receive bupivacaine or placebo. Assessment of postoperative pain using a numerical scoring system was made by the child, the recovery sister and the ward sister, and by measurement of the time between the end of the operation and the need for narcotic injection. Analysis revealed significantly less pain (P < 0.03) in the immediate postoperative period in the treated group. The use of bupivacaine wound infiltration as a supplement to other current methods of pain control is recommended. PMID- 8428268 TI - Simultaneous dynamic electromyographic proctography and cystometrography. AB - The rectum and bladder share a common origin and work in harmony; disturbance in one may lead to a similar problem in the other. The two have, however, not previously been investigated dynamically and simultaneously. A new system has been devised allowing dynamic integrated electromyographic proctography to be performed simultaneously with cystometrography in 41 patients (28 women and 13 men). Of 16 women with chronic constipation who underwent the combined study, ten were shown to have obstructed micturition. Eight of these ten women exhibited an inappropriate contraction of the puborectalis muscle during micturition, compared with one of the six with normal micturition (P = 0.02). Of the patients with obstructed micturition, seven of eight women with a rectocele also had anismus, compared with none of four women with a rectocele among those who were able to micturate normally (P = 0.02). Eight of the ten women with obstructed micturition thus had a proctological abnormality that explained the urological symptoms. Of 12 women with idiopathic faecal incontinence who underwent the combined study, eight were shown to have genuine stress incontinence of urine. In seven of these the severity was shown to be of type 2a or greater, indicating that pelvic floor dysfunction may be the causal factor of both rectal and urinary symptoms. The combined study shows abnormalities in one system that could explain similar symptoms in the other. PMID- 8428269 TI - Effect of anorectal eversion during restorative proctocolectomy on anal sphincter function. AB - Twenty-six patients underwent restorative proctocolectomy with end-to-end ileal pouch-anal anastomosis, without resection of the anal mucosa, by the eversion technique. Before surgery patients underwent tests of anorectal function. These were repeated a median of 8 (range 3-21) months after operation. The median (range) maximum resting anal pressure was 93 (36-149) cmH2O before and 71 (40 131) cmH2O after operation (P = 0.002). The median (range) maximum squeeze pressure before operation was 136 (73-280) cmH2O; it was 149 (69-290) cmH2O afterwards (P not significant). The median (range) length of the anal sphincter was 3.5 (2.5-4.5) cm before and 3.5 (2.0-4.5) cm after operation (P not significant). Thresholds for sensation in the upper, middle and lower thirds of the anal canal before and after operation were, respectively, 8.7 versus 8.7, 6.8 versus 7.4 and 4.2 versus 6.2 mA (P not significant). All 26 patients were continent, although one experienced minor leakage. Function of the anal sphincter is not significantly impaired by eversion of the rectum and anus during restorative proctocolectomy. PMID- 8428270 TI - Breast cancer in Nigerian women. PMID- 8428271 TI - Laparoscopic splenectomy. PMID- 8428272 TI - Laparoscopic pyloroplasty for duodenal ulcer. PMID- 8428273 TI - Laparoscopic repair of perforated peptic ulcer. PMID- 8428274 TI - Total dysphagia from intramural haematoma following sclerotherapy for oesophageal varices. PMID- 8428275 TI - Perforation of the bowel by suction drains. PMID- 8428276 TI - Meconium ileus: a review 1972-1990. PMID- 8428277 TI - Perforation of the bowel by suction drains. PMID- 8428278 TI - Vascular inflow exclusion and hepatic resection. PMID- 8428279 TI - Primary versus staged resection for acute obstructing colorectal carcinoma. PMID- 8428280 TI - Early gastric cancer. PMID- 8428281 TI - Contemporary management of the appendiceal mass. AB - An appendiceal mass is the end result of a walled-off appendiceal perforation and represents a pathological spectrum ranging from phlegmon to abscess. Over the past decade, improved imaging and interventional radiological techniques have allowed a more accurate definition of pathology and thus a more specific and less invasive management than was previously possible. A management policy should be possible that allows over 80 per cent of patients presenting with an appendiceal mass to be safely spared an open operation. PMID- 8428282 TI - Congenital arteriovenous malformations. PMID- 8428283 TI - Intraoperative streptokinase: a useful adjunct to balloon-catheter embolectomy. AB - Intraoperative thrombolysis was attempted in 31 acutely ischaemic legs after operative arteriography had demonstrated residual distal thrombus or occlusion following balloon-catheter thromboembolectomy. There were 30 patients, 16 men and 14 women, aged 43-82 (median 73) years. The indication for operation was severe ischaemia with sensorimotor loss in 25 limbs, failed percutaneous thrombolysis in three and acute graft occlusion in three. A total of 21 perfemoral, 11 perpopliteal and four graft embolectomies were initially performed. Following arteriography, 100,000 units streptokinase was infused down the isolated distal arterial tree over 30 min and arteriography repeated. Complete lysis was achieved in 11 legs (35 per cent) and partial lysis in 12 (39 per cent). Additional procedures required included six operative angioplasties and six bypass grafts. After operation pedal pulses were restored in 14 limbs (45 per cent), with a viable leg in 23 cases (74 per cent) at the time of patient discharge or death. There were five wound haematomas but no evidence of systemic fibrinolysis. Four amputations were required, none in the group undergoing successful lysis, and there were seven deaths, five from cardiac disease. Arteriography after balloon catheter embolectomy is essential to detect residual thromboembolus and intraoperative streptokinase appears to be a safe and effective way of removing this. PMID- 8428284 TI - Choice of agent for peripheral thrombolysis. AB - Evidence has been accumulating that tissue plasminogen activator (tPA) is a more rapid and effective agent than streptokinase for peripheral thrombolysis. Twenty three patients with acute limb-threatening ischaemia treated with tPA (0.5 mg h 1) over 15 months were compared with 20 consecutive patients previously receiving streptokinase (5000-10,000 units h-1). There were no major differences between the rates of complete and partial lysis (61 per cent for tPA versus 65 per cent for streptokinase) or limb salvage (65 versus 55 per cent respectively). Complication rates were also similar. It was not possible to show that tPA, an agent ten times more expensive than streptokinase, was superior for peripheral thrombolysis. PMID- 8428285 TI - Myocardial infarction after reconstruction of the abdominal aorta. AB - The records of 555 patients who underwent elective abdominal aortic reconstruction were analysed to identify risk factors predisposing to cardiac complications. Perioperative myocardial infarction occurred in 35 patients (6.3 per cent), of whom 12 (2.2 per cent) died. Using multiple logistic regression, four preoperative factors that were independently predictive for postoperative myocardial infarction were identified with the following relative risk ratios: history of transient ischaemic attacks, 5.08; raised serum creatinine level, 3.66; age > 60 years, 3.00; and angina requiring regular treatment, 2.74. A scoring system using these four factors was devised that identified 150 of the 555 patients as at high risk, and included 69 per cent of those with myocardial infarction after operation and 83 per cent of those who died. A clinical risk score is a practical first step in identifying patients at risk of myocardial infarction; it would also reduce the number of patients for whom sophisticated tests of cardiac function were necessary. PMID- 8428286 TI - Autotransfusion in aortic surgery: the Haemocell System 350 cell saver. AB - Autotransfusion was performed during elective abdominal aortic reconstruction in ten patients using the recently developed Haemocell System 350. A mean of 60 per cent of total blood loss was salvaged, and during operation each patient was autotransfused 1 unit. Good preservation of cellular components, including platelets, was seen after processing with the device, which uses a vortex mixing filter for cell separation. There was no evidence of coagulopathy; mean free plasma haemoglobin levels were slightly raised (17 mg/dl) and plasma heparin concentration was negligible (0.10 units/ml) 4 h after surgery. A transient drop in plasma fibrinogen levels and the appearance of fibrin degradation products in low concentrations (mean 1.5 mg/l) were seen. Oxyhaemoglobin dissociation curves showed the salvaged red blood cells to have a normal affinity for oxygen. Renal and hepatic function remained unaltered and there were no complications attributable to autotransfusion. The device was easy to handle and a trained operator was not required. PMID- 8428287 TI - Ankle fracture is associated with prolonged venous dysfunction. AB - Leg swelling is a significant problem following ankle fracture. Venous pump function and femoral and popliteal venous patency were assessed prospectively in 26 patients with ankle fractures requiring open reduction, internal fixation and immobilization in plaster. Functional venous volume, venous filling index (VFI), ejection volume fraction (EVF) and residual volume fraction (RVF) were measured using air plethysmography at 5 days and at 6, 12 and 18 weeks after fracture. The uninjured leg was used as a control. Popliteal and femoral venous patency was determined using duplex ultrasonography. No patient developed deep vein thrombosis during the study. At 5 days after fracture there was a significant reduction in mean(s.d.) EVF, 18.2(12.1) versus 55.9(19.5) per cent, and increase in RVF, 87.0(14.3) versus 42.5(22.2) per cent (both P < 0.001). Analogous values were similar at 6 weeks, EVF 28.5(21.2) versus 55.6(21.9) per cent, RVF 82.2(16.8) versus 48.5(23.8) per cent (both P < 0.001), and at 12 weeks, EVF 39.1(16.0) versus 60.3(14.9) per cent, RVF 64.7(18.8) versus 38.8(13.2) per cent (both P < 0.001). However, by 18 weeks there was no significant difference in venous function between fractured and control limbs. It is concluded that there is a significant and prolonged impairment in venous pump function following ankle fracture. PMID- 8428288 TI - Perioperative nutritional support. PMID- 8428289 TI - Parathyroidectomy in the treatment of patients with chronic renal failure: a 10 year review. AB - The technique and outcome of surgery for the manifestations of secondary hyperparathyroidism are described. Forty-seven patients were analysed retrospectively over a 10-year period. Total parathyroidectomy and autotransplantation to the forearm was performed in all but five patients. Improvement occurred in 87 per cent of patients and there were no graft failures. Of six deaths, none was related to surgery. Four patients had recurrent hyperparathyroidism, three being graft dependent and one requiring re-exploration of the neck. Total parathyroidectomy and autotransplantation is a safe and effective approach. The major management problem was graft hyperplasia, seen in three of the 47 patients and managed by simple excision. PMID- 8428290 TI - Timing of surgery for breast cancer in relation to the menstrual cycle and survival of premenopausal women. PMID- 8428291 TI - Centroinferior hemimastectomy. PMID- 8428292 TI - Outcome 5 years after 360 degree fundoplication for gastro-oesophageal reflux disease. AB - Forty patients with a mean age of 45 (range 22-65) years were operated on between 1982 and 1985 for gastro-oesophageal reflux disease with a short floppy 360 degree fundoplication. The results of the operation were determined by endoscopy, oesophageal manometry, ambulatory 24-h pH recording and symptom evaluation 6 months and 5 years after operation. These results were compared with findings in healthy controls. The median pressure in the lower oesophageal high-pressure zone was 13.3 (interquartile range (i.q.r.) 11.3-21.3) mmHg after 5 years, which did not differ significantly from the value at 6 months' follow-up or from that in controls. It was, however, significantly higher than the preoperative pressure. The median intra-abdominal length of the high-pressure zone was 1.7 (i.q.r. 1.3 2.3) cm after 5 years, significantly less than at 6 months but equal to control length. Measurement of the proportion of total time at pH < 4 at 5 years (median 0.2 (i.q.r. 0.0-0.6) per cent) and 6 months after operation revealed a significant reduction in acid reflux compared with preoperative values and normal controls. There was no significant difference in acid exposure between the two postoperative investigations. Endoscopy showed that 27 patients had no oesophagitis, three had erythema and three persistent Barrett's oesophagus 5 years after operation. Normal belching was possible in 22 patients and 18 experienced increased flatulence 5 years after fundoplication. An independent gastroenterologist found that the result was excellent in 16 patients, good in 16 and fair in four; two patients had a poor overall outcome of the operation. It is concluded that a 360 degree fundoplication provides good long-term control of reflux and that slight symptoms of overcompetence are common among patients operated on without affecting the overall result. PMID- 8428293 TI - Cholecystectomy and oesophageal reflux: a prospective evaluation. AB - The effect of cholecystectomy on oesophageal function was examined prospectively. Of 37 patients studied, 17 (46 per cent) were still symptomatic 3-4 months after surgery. Thirteen patients (35 per cent) had abnormal oesophageal pH profiles before cholecystectomy, increasing to 27 (73 per cent) afterwards (P < 0.002). The mean (s.e.m.) DeMeester acid reflux score increased from 15.2(2.1) to 34.2(5.2) (P < 0.001) after operation. Lower oesophageal sphincter function, as measured by the sphincter function index, was significantly reduced in the patients with abnormal pH profiles after operation (P < 0.01). Mean(s.e.m.) supine gastric alkaline shift (proportion of time at pH > 4) increased from 9.2(2.0) to 17.7(3.7) per cent (P < 0.02) and the incidence of gastritis from eight patients (22 per cent) to 23 (62 per cent) (P < 0.001). These data suggest that cholecystectomy results in gastro-oesophageal reflux that appears to be related to compromised sphincter competence. PMID- 8428294 TI - Gastric remnant cancer as a metachronous multiple lesion. AB - The pathological characteristics and natural history of 35 gastric remnant cancers after partial gastrectomy for a malignant condition and 16 gastric cancers after gastrectomy for benign conditions were compared. Gastric remnant cancer following malignant disease was characterized by a well defined rather than diffuse appearance (in 43 versus 12 per cent of gastric remnant cancers after benign conditions, P < 0.05), location away from the anastomosis (83 versus 25 per cent, P < 0.05) and a shorter interval after the first operation (5-14 versus > or = 20 years, P < 0.01). The 15- and 16-year survival rates appeared to be worse for gastric remnant cancer after malignant than after benign disease, but there was no statistically significant difference between the two groups in a generalized Wilcoxon test. In the former, direct invasion to adjacent organs was frequently observed. These findings suggest that gastric remnant cancer after gastrectomy for malignancy may be a metachronous multiple lesion, while that following benign disease may occur as a new cancer caused by the partial gastrectomy. To improve the survival of patients with cancer after gastrectomy for malignancy, a rational extended operation may be useful. PMID- 8428295 TI - Nucleolar organizing regions in the operated rat stomach: relationship to metaplasia, dysplasia and carcinoma. AB - This study investigated the role of duodenogastric reflux in rats after antecolic gastrojejunostomy. At the end of the study, adenocarcinoma was present in 43 per cent, dysplasia in 14 per cent and intestinal metaplasia in 43 per cent of animals. No such changes were found in control rats. A technique for silver staining nucleolar organizing regions (AgNORs) was applied to these lesions and to the control group. The AgNOR count gradually increased from normal gastric mucosa to carcinoma. This technique demonstrated differences in AgNOR count between normal mucosa and other lesions (adenocarcinoma, dysplasia, intestinal metaplasia) (P < 0.001). There was, however, considerable overlap among adenocarcinoma, dysplasia and metaplasia (P > 0.1). PMID- 8428296 TI - Medullary thyroid carcinoma. AB - Medullary thyroid carcinoma accounts for 5-10 per cent of thyroid malignancies and occurs sporadically, and in three familial settings with autosomal dominant inheritance. Calcitonin, a 32 amino acid 3.5-kDa protein, is a biochemical marker of the tumour. Serum levels correlate with disease burden. Treatment initiated by screening allows disease removal at a premalignant phase: C cell hyperplasia. Genetic linkage studies locate the multiple endocrine neoplasia type 2A gene near the centromere of chromosome 10. Flanking genetic markers may allow polymorphic probes to examine DNA from a once-only blood sample to identify affected individuals. At what age thyroidectomy should be carried out in these patients is unclear. PMID- 8428297 TI - Transit disorders of the gastric remnant and Roux limb after Roux-en-Y gastrojejunostomy: relation to symptomatology and vagotomy. AB - Patients after Roux-en-Y gastrojejunostomy frequently complain of upper abdominal pain, fullness, nausea and vomiting. This study was performed to clarify the relationship of this Roux-en-Y syndrome to transit disorders in the gastric remnant and Roux limb, and to vagal status. Using a questionnaire, 35 of 66 patients operated on between 1976 and 1987 were judged to suffer from the Roux syndrome. Gastrojejunal transit was studied by scintigraphy with a solid test meal in 61 patients, 34 of whom were symptomatic. The median (interquartile range) gastric half-emptying time was longer in symptomatic than in asymptomatic patients (79 (43-146) versus 56 (27-79) min, P < 0.05), and in patients with a bilateral vagotomy than in those without a vagotomy (94 (43-225) versus 59 (31 77) min, P < 0.05). Stasis in the Roux limb was observed in 18 of 28 symptomatic and in only three of 27 asymptomatic patients (P < 0.01). The median (interquartile range) fraction of activity emptied from the stomach and remaining in the Roux limb at 60 min was 54 (39-60) per cent in symptomatic patients and 33 (21-40) per cent in those without symptoms (P < 0.01). Stasis in the Roux limb was not related to vagal status. No relationship between slow gastric emptying and Roux-limb stasis was found. Slow gastric emptying, Roux-limb stasis or a combination of both was found in 30 of 34 symptomatic and in only nine of 27 asymptomatic patients (P < 0.01). It is concluded that both slow gastric emptying and Roux-limb stasis can be interpreted as causing the Roux syndrome. Vagotomy seems to be the major cause of slow gastric emptying, but it is not related to stasis in the Roux limb. PMID- 8428298 TI - Omental patching of a large perforated duodenal ulcer: a new method. PMID- 8428299 TI - Bacterial translocation in acute liver failure induced by 90 per cent hepatectomy in the rat. AB - Bacterial infection and bacteraemia have been observed in patients with acute liver failure. The exact source of bacteria and nature of pathophysiological mechanisms explaining the development of infection remain unclear. In the present study, acute liver failure was induced by 90 per cent hepatectomy in the rat. The mesenteric lymph nodes and organs were harvested aseptically for bacteriological culture after sham operation or 90 per cent hepatectomy. Function of the liver and reticuloendothelial system (RES) was assayed; gut oxygen extraction was also measured. Translocation of enteric bacteria occurred 2 h after operation and increased with time following hepatectomy. Overgrowth of Escherichia coli in the distal small intestine started 2 h after operation. RES function decreased immediately after 90 per cent hepatectomy; uptake rates per gram tissue in other organs increased significantly. These results indicate that bacterial translocation occurred early after 90 per cent hepatectomy, associated with a decrease in RES function and gut oxygen extraction, and overgrowth of intestinal bacteria. PMID- 8428300 TI - Histamine but neither angiotensin nor vasopressin increases antibody uptake into xenograft colorectal liver metastases. AB - Although the majority of colorectal carcinomas express carcino-embryonic antigen (CEA), systemic anti-CEA antibody administration is an ineffective treatment for colorectal liver metastasis. A xenograft model of human colorectal carcinoma in the rat was used to determine anti-CEA antibody uptake into liver metastases. The influence of systemic (iliolumbar vein) or regional (gastroduodenal artery) delivery and effects of regional delivery of histamine, angiotensin II and vasopressin on anti-CEA antibody uptake by metastases were examined. Systemic antibody delivery achieved a median tumour:liver antibody uptake ratio of 1.60 (interquartile range (i.q.r.) 1.02-2.51). Regional delivery resulted in a similar median ratio of 1.61 (i.q.r. 1.22-2.46). Histamine and antibody delivered regionally produced a median tumour:liver ratio of 3.15 (i.q.r. 2.50-4.27), which was significantly greater than that obtained with systemic delivery (P = 0.004). Regional infusion of angiotensin resulted in a median (i.q.r.) ratio of 2.23 (1.58-2.49) and vasopressin in 2.15 (1.41-2.60), values that were not significantly different from those found with systemic or regional delivery alone. When both angiotensin and histamine were infused with antibody, the median tumour:liver ratio was 3.09 (i.q.r. 2.22-4.31), significantly greater than for systemic delivery (P = 0.01) but not significantly different from that obtained following the addition of histamine alone (P = 0.94). Histamine significantly increases antibody uptake in a model of liver metastasis and may improve the effectiveness of targeted immunotherapy in the treatment of colorectal liver metastasis. PMID- 8428301 TI - Controlled liver splitting for transplantation in two recipients: technique, results and perspectives. AB - A technique of controlled liver splitting for transplantation in two recipients is proposed, based on a full anatomical assessment of the graft including arteriography and cholangiography on the back-table. Using eight livers, 16 patients received a graft: right liver (eight patients), left lobe (four) or left liver (four). Twelve patients required urgent or very urgent transplantation. Anatomical assessment of the graft demonstrated a portal bifurcation in all cases, a common trunk of the left and middle hepatic veins in five, a right biliary duplication in three and duplication of the left branch of the middle hepatic artery in one. After revascularization of the graft, bleeding was greater in patients with a right graft, particularly if the middle hepatic vein had been ligated. The main postoperative complications were hepatic artery thrombosis (four cases), biliary complications (four), portal vein thrombosis (two), haematoma (two) and abscess (two). No primary non-function of the graft was observed. The postoperative survival rate was 75 per cent. The four patients in whom transplantation was not considered urgent are still alive. The immediate survival rate of the grafts was 69 per cent. These results compare favourably with those in the literature. In spite of the technical, logistical and ethical problems raised by this technique, the results suggest that controlled liver splitting for transplantation in two recipients may in the future significantly improve the feasibility of liver transplantation. PMID- 8428302 TI - Liver transplantation in patients with thrombosis of the portal, splenic or superior mesenteric vein. AB - Fourteen cases are presented of preoperative portal vein thrombosis complicating orthotopic liver transplantation from an experience of 195 transplants carried out between April 1986 and April 1991. In four patients who suffered rethrombosis of the portal vein, the mortality rate was 100 per cent from various causes. Overall there were six deaths; two of those who died had a patent portal vein at death. Three patients underwent retransplantation: one for primary non-function, two for rejection. It is concluded that: (1) portal vein thrombosis should not represent an absolute contraindication to liver transplantation; (2) preoperative screening of prospective transplant recipients for portal thrombosis should be routine; (3) postoperative anticoagulation therapy and periodic Doppler ultrasonographic assessment of portal vein flow are important elements of post transplant management; and (4) with thrombectomy and portal vein resection an end to-end portal anastomosis may be performed with good results. PMID- 8428303 TI - Management of traumatic liver injuries. AB - Liver injuries in Europe are usually caused by blunt trauma and a high mortality rate is generally reported. The severity of liver injury in 175 patients was graded from I to V and the Injury Severity Score assessed. Seventy-five patients in a prospective study (1987-1990) were treated according to a defined protocol. Non-operative management was used for those who were haemodynamically stable on admission. In unstable patients who proceeded to surgery, liberal use of packing was made and a low threshold for relaparotomy employed. Increasing experience resulted in fewer indications for resection and a 40 per cent rate of non operative treatment in the prospectively managed group. The overall mortality rate was 12 per cent (15 per cent in the retrospective and 8 per cent in the prospective group). Death in patients with multiple injuries should only rarely result from liver trauma. PMID- 8428304 TI - Traumatic pancreatic pseudocysts. AB - Fifteen patients who developed pseudocysts following pancreatic trauma were evaluated to determine outcome in relation to the nature and site of pancreatic duct injury. Pseudocysts developed in eight patients operated on within 48 h of abdominal trauma and in seven who were initially treated conservatively. In none was duct injury diagnosed during initial management. Presentation was a median of 20 (range 8-360) days after injury. In 14 patients, pseudocysts (mean diameter 9 (range 3-16) cm) were confirmed by computed tomography or ultrasonography. Endoscopic retrograde pancreatography (ERP) demonstrated the site and severity of the duct injury in eight of 11 patients. Two patients with side duct injury on ERP were treated successfully without intervention. Pseudocysts arising from distal duct injuries (four patients) were treated by percutaneous aspiration or catheter drainage, although one patient required subsequent distal resection for recurrent pancreatitis caused by a pancreatic duct stricture. Three patients with duct injuries in the neck or body with pancreatic disruption underwent distal pancreatectomy. Proximal duct injuries with mature pseudocysts (three patients) were drained internally. Three patients had complicated pseudocysts (haemorrhage in one, sepsis in two) that necessitated emergency laparotomy and external drainage; one of these patients died from sepsis. These findings suggest that traumatic pancreatic pseudocysts that follow peripheral duct injury may resolve spontaneously, whereas those associated with distal duct injuries can be treated by percutaneous aspiration or catheter drainage. Proximal duct injuries, however, require surgical intervention using either resection or internal drainage, depending on the maturity of the cyst wall. PMID- 8428305 TI - Symptomatic wandering spleen. PMID- 8428306 TI - Assessment of main pancreatic duct integrity by endoscopic retrograde pancreatography in patients with acute pancreatitis. AB - The integrity of the main pancreatic duct (MPD) was evaluated by endoscopic retrograde pancreatography (ERP) in a retrospective study of 105 patients with acute pancreatitis presenting over an 11-year period (1980-1991). The findings were compared with clinical outcome and the need to operate for local pancreatic complications. Patients were divided into two groups. Group 1 (n = 89) had either clinically mild pancreatitis or severe disease but no surgery for local complications, and < 25 per cent necrosis on contrast-enhanced computed tomography (CT) (n = 48). Group 2 patients (n = 16) had clinically severe pancreatitis and underwent surgery for local complications and/or had > or = 25 per cent necrosis on CT (n = 12), at surgery or post mortem. All 89 patients in group 1 had an intact MPD at ERP, which was performed a median of 6 (range 0-30) days after onset of pancreatitis; the median age was 63 (range 20-88) years and there were no deaths. The median age of patients in group 2 was 59 (range 26-85) years. ERP in this group was performed in four patients < 5 days after onset and all had an intact MPD; one died with necrosis and another from a cerebrovascular accident. ERP was performed > or = 5 days after onset in the other 12 patients; five had an intact MPD and two required surgery for pseudocyst drainage only; seven had a disrupted MPD and all required surgery for pancreatic necrosis (one death). It is concluded that an intact MPD was a feature of mild pancreatitis, whereas disruption occurred > 4 days after onset in patients with necrosis necessitating surgery. PMID- 8428307 TI - Lexical representation and morpho-syntactic parallelism in the left hemisphere. AB - This experiment investigated the sensitivity of the left and right cerebral hemispheres to morphologically ambiguous verbs (i.e., chased) and to morphologically unambiguous verbs (i.e., stolen) in a divided visual-field paradigm with normal subjects. Lexical decisions made to the ambiguous verbs were faster than those to the unambiguous verbs only when presented in the right visual field. The results suggest that the traditional notion of left-hemispheric specialization for syntactic processing can be extended to the retrieval of lower level morphological representations. PMID- 8428308 TI - Cortical differences in tonal versus vowel processing as revealed by an ERP component called mismatch negativity (MMN). AB - Event-related potentials were recorded from four aphasic subjects in order to study if discrimination of synthetic vowels is impaired by left posterior brain damage. A component called the mismatch negativity (MMN) which is assumed to reflect basic discriminatory processes of auditory stimuli was measured. In accordance with the hypothesis, two patients with posterior lesions failed to show any MMN response to synthetic vowels, whereas two patients with predominantly anterior lesions produced the response. The fact that all four patients produced an MMN response to sine wave stimuli indicates that the result does not reflect an across-the-board effect. PMID- 8428309 TI - The structural determinants of recovery in Wernicke's aphasia. AB - Recovery of comprehension and total language in 22 Wernicke's aphasics was correlated with lesion size and extent of involvement of certain structures on CT. Recovery rates and outcomes were separately examined using 0-3 months and 0 12 months poststroke language data. Quantitative measures of structural damage were regressed on total aphasia and comprehension outcome measures. Supramarginal and angular gyri appeared to be the most significant structures in recovery in addition to initial severity and lesion size. This was confirmed by using ANOVA to compare the extent of involvement in each postcentral structure among the poor, moderate, and good recovery groups. The superior temporal and middle temporal gyri are less involved in the good recovery group. Structures posteriorly adjacent to Wernicke's area are important for compensation in Wernicke's aphasia and in the accompanying comprehension deficit. Persisting Wernicke's aphasia usually involves the supramarginal and angular gyri in addition to the superior temporal area. PMID- 8428310 TI - Language deviations in aphasia: a frequency analysis. AB - Thirty right-handed left hemisphere-damaged patients were taken and divided into five groups (transcortical motor, Broca, conduction, Wernicke, and anomic aphasia). Language deviations were scored and analyzed for the Picture Description (Plate No. 1, The Cookie Theft), Repetition (Words, High and Low Probability Sentences), and Naming (Responsive Naming, Confrontation, and Body part naming) subtests of the Boston Diagnostic Aphasia Examination--Spanish version (Goodglass & Kaplan, 1979). A classification of paraphasias is proposed. Language deviations were scores for the following groups: Literal paraphasias (phoneme omissions, additions, displacements, and substitutions), verbal paraphasias (formal, morphologic, semantic, and unrelated), syntagmatic paraphasias, circumlocutions (object description and instrumental function), indefinite anaphors, and neologisms. Frequency of different types of language deviations is presented in the five aphasia groups. It was found that some paraphasic errors appeared in several aphasia groups; others were characteristics of specific aphasic syndromes. PMID- 8428312 TI - Verbal recognition memory performance in unilateral temporal lobe epilepsy. AB - Recognition memory performance of 91 left and right unilateral temporal lobe epilepsy (TLE) patients was examined prior to and post anterior temporal lobectomy. The left TLE group showed more impaired response discrimination ability than the right TLE group; however, their false positive rate varied as a function of distractor type. Prior to temporal lobe resection, false positives were higher for semantically related distractors (novel or from an interference list) than for semantically unrelated items from an interference list, phonemic distractors, or unrelated errors. Following temporal lobe resection, only novel semantically related distractors in the left TLE group showed a significant increase in false positives, and they were also significantly higher than all other error types. The results of this study suggest that the left TLE patients have impaired individual item recollection in the context of intact information about basic semantic attributes. PMID- 8428311 TI - Visual recognition in right brain-damaged patients: evidence from a tachistoscopic confrontation naming task. AB - Right brain-damaged patients performed a confrontation naming task. Pictures were tachistoscopically presented to the right visual field and selected for their high degree of canonicity and name agreement measured in control subjects. Compared to controls, patients exhibited significant errors. Misnamings were mainly perceptual, i.e., visual-semantic or purely visual errors. No evidence of pure semantic error was found. Half of visual-semantic misnamings referred to objects of a different size (scale error) within the correct semantic field. Misnamings are tentatively attributed to a disturbance of the visual-imagistic automatic encoding process at the level of the pictorial data store. PMID- 8428313 TI - Progressive aphasia without dementia--a clinical and cognitive neuropsychological analysis. AB - This article provides a clinical and cognitive neuropsychological analysis of a patient who presented a marked and progressive linguistic disturbance in the absence of any other significant cognitive impairment. A PET scan indicated that this disorder was associated with focal left temporofrontal hypometabolism. The essential characteristics of this disorder were anomia, surface dyslexia, and surface dysgraphia, along with a mild grammatical disturbance and deficits in both visual and auditory word comprehension. The patient's disorder is considered in relation to other similar cases and to modular accounts of acquired linguistic disorders. PMID- 8428314 TI - Retrieval inhibition in anomia. AB - Reading latencies for anomic temporal lobe epileptics, nonanomic epileptic patient controls, and normal controls were measured in semantic priming paradigms. Both the epileptic controls and the normal controls showed typical semantic facilitation with faster response times following related than unrelated primes. The anomic subjects, on the other hand, were much slower to read targets following presentation of semantically related items than following unrelated primes. This inhibition effect was seen to increase as the number of related primes increased. These patterns were observed both when picture primes (Experiment 1) and word primes (Experiment 2) were used. These findings were interpreted as evidence for a category-specific retrieval inhibition in the anomic epileptic subjects. PMID- 8428315 TI - Bedside/point-of-care technology: the 'window' into the integrated clinical database. PMID- 8428316 TI - FLORENCE: synthesis of case-based and model-based reasoning in a nursing care planning system. AB - In this article, the data structures and reasoning methodologies of FLORENCE, an expert system that advises on the identification of nursing diagnoses in a new client, are described. FLORENCE is primarily a case-based reasoning system, but has an ancillary model-based reasoning system to advise on complex problems. The use of multiple reasoning methods allows for better simulation of the reasoning process of the expert clinician. PMID- 8428317 TI - HyperCard to SPSS: improving data integrity. AB - This article describes a database design that captures responses in a HyperCard stack and moves the data to SPSS for the Macintosh without the need to rekey data. Pregnant women used an interactive computer application with a touch screen to answer questions and receive educational information about fetal alcohol syndrome. A database design was created to capture survey responses through interaction with a computer by a sample of prenatal women during formative evaluation trials. The author does not compare this method of data collection to other methods. This article simply describes the method of data collection as a useful research tool. PMID- 8428318 TI - The relationship between performance on computer-based clinical simulations and two written methods of evaluation: cognitive examination and self-evaluation of expertise. AB - The purpose of this study was to explore the relationship between computer-based clinical simulation performance and two commonly used methods of evaluation: cognitive examination and self-evaluation of expertise. The convenience sample of 68 critical care nurses completed the Basic Knowledge Assessment Tool for Critical Care (BKAT), a cardiovascular self-assessment tool (CST), and two computer-based clinical simulations requiring the management of myocardial infarction patients with tachydysrhythmias (atrial flutter and ventricular tachycardia). Simulation performance was measured by proficiency score and patient outcome (live/die). Proficiency on the atrial simulation was significantly correlated with knowledge as measured by the BKAT (r = 0.46, p 0.001) and with self-evaluation of clinical expertise (r = 0.32, p = 0.008). Atrial patient outcome was also significantly correlated with the BKAT (r = 0.28, p = 0.006) and the CST score (r = 0.22, p = 0.036). However, in the ventricular simulation only proficiency score and self-evaluation of expertise were significantly correlated (r = 0.24, p = 0.049). The modest relationships found in this study may indicate that, while clinical simulation performance is related to knowledge and self-evaluation of expertise, the constructs measured are not synonymous. This evidence provides support for the use of clinical simulations as an adjunct to other methods of evaluation. PMID- 8428319 TI - Nursing information systems in the year 2000: another perspective. PMID- 8428320 TI - EXTEND: a prototype expert system for teaching nursing diagnosis. AB - In this article, the development and uses of EXTEND (Expert system for Education in Nursing Diagnosis), a computer assisted instruction package for teaching nursing diagnosis using the categories proposed by the North American Nursing Diagnosis Association, are described. EXTEND provides a knowledge base that is able to deduce a nursing diagnosis from case factors and extends the normal use of expert systems beyond diagnosis to diagnosis education by providing answers to student questions about a simulated client and by guiding them through the diagnostic task. Furthermore, EXTEND allows the teacher to record the steps taken by students in the assessment and diagnosis of the simulated client, and use this as the focus of further discussion. The system is a response to the problems of teaching nursing diagnoses cited in the professional literature. PMID- 8428321 TI - Bedside/point-of-care technology: this issue's featured vendors. PMID- 8428322 TI - Robo-Pill. PMID- 8428323 TI - Adjuvant therapy for localized prostate cancer. AB - BACKGROUND: In 1978, the National Prostatic Cancer Project launched two protocols evaluating adjuvant therapy after surgery (Protocol 900) or irradiation (Protocol 1000) for clinically localized prostate cancer. All patients underwent staging pelvic lymphadenectomy. METHODS: After definitive treatment, the patients were randomized either to receive cyclophosphamide 1 g/m2 intravenously every 3 weeks for 2 years or estramustine phosphate 600 mg/m2 orally daily for up to 2 years or to undergo observation only. Patient accession closed in 1985 and includes 184 patients in Protocol 900 (170 evaluable) and 253 in Protocol 1000 (233 evaluable). RESULTS: Lymph node involvement was identified in 198 patients (49% of total), 29% in Protocol 900 and 63% in Protocol 1000. The median progression free survival (PFS) and survival were greater for patients in Protocol 900 compared with 1000, regardless of the adjuvant therapy. This reflected the greater proportion of patients with lower pathologic stage disease in the surgically treated group. The median PFS was significantly greater for all patients in Protocol 1000 receiving estramustine phosphate adjuvant (48.2 months) compared with patients randomized to receive cyclophosphamide (35.6 months). The median PFS for patients with nodal involvement in Protocol 1000 who received estramustine phosphate adjuvant was prolonged significantly (37.3 months) compared with no treatment (20.9 months). The median PFS for patients with limited nodal disease in Protocol 1000 was longer (39.9 months), regardless of the adjuvant therapy, compared with those with extensive nodal disease (20.7 months). However, in the latter patient group, those receiving adjuvant estramustine phosphate had a significantly longer median PFS (32.8 months) compared with those receiving adjuvant cyclophosphamide (22.7 months) or no adjuvant therapy (12.9 months). CONCLUSION: Adjuvant estramustine phosphate was beneficial in patients with prostate cancer and pelvic node involvement who received irradiation as definitive treatment. PMID- 8428324 TI - Counseling the patient with regional metastasis of prostate cancer. AB - BACKGROUND: Treatment options for all stages of prostate cancer are a source of considerable debate; patients with regional nodal metastases have been advised to undergo a wide range of therapies from observation with delayed endocrine treatment to an aggressive combination of radical prostatectomy (RP), radiation therapy (RT), and hormonal treatment. RESULTS: In the absence of reliable data to confirm the wisest choice, a clinician must counsel the patient based on treatment morbidity, the best estimate of efficacy, and an appreciation of the patient's individual desires regarding treatment. Patients with minimal microscopic nodal metastases treated with RP are destined to have relapses ultimately, but the development of metastases may take 5-10 years even with no additional therapy. Immediate hormonal treatment combined with RP currently is used more commonly, and a subgroup of patients have remained disease-free for more than 10 years. Local morbidity can occur as a consequence of RP or RT or, later in the disease course, secondary to progressive growth of the cancer in the prostate. Finally, the ability to detect nodal metastases with minimal morbidity is available using laparoscopic pelvic lymphadenectomy. Thus, more emphasis may be placed on defining nodal status before definitive therapy. CONCLUSIONS: Historically, nodal status was used to define whether the patient was eligible for aggressive local therapy; the presence of positive lymph node metastases disqualified a patient to receive RP or RT. In the future, nodal status may not define local therapy but instead may determine whether the patient receives adjuvant hormonal therapy or not. This more aggressive approach has gained acceptance because of the lowered morbidity of both RP or RT and the extended disease-free survival in at least a segment of the patient population. Because patients initially often select the form of local therapy best suited for them, this approach respects this initial decision by offering additional psychologic support in the long term. PMID- 8428325 TI - Sexual rehabilitation after treatment for prostate cancer. AB - Although there is little evidence that sexual behavior causes prostate cancer, men with prostate cancer often have sexual dysfunction before the cancer diagnosis is made. Each treatment for prostate cancer increases the prevalence of sexual problems. After nerve-sparing radical prostatectomy, the chance of recovering erections is better for men who are younger and in whom both neurovascular bundles can be spared. Definitions of "potency" after nerve-sparing surgery have not specified the rigidity of the erections achieved. Thus, some men classified as "potent" may wish additional sexual rehabilitation. The chance that definitive radiation therapy will cause erectile dysfunction probably has been overestimated. The prevalence rate may be closer to 25% of men with new problems compared with the 50% often cited in the literature. Men are more at risk to have erection problems after radiation therapy if the quality of erections before treatment was borderline. Hormonal therapy has an impact on the central mechanisms mediating sexual desire and arousability. Therefore, with most treatment methods, only approximately 20% of men remain sexually functional. Newer antiandrogenic drugs interfere less with sexual function, but their long term ability to control prostate cancer is still under investigation. Sexual rehabilitation should be addressed by the primary care team. Sexual partners should be included in brief sexual counseling, even when a mechanical treatment for erectile dysfunction is prescribed. PMID- 8428326 TI - Antiandrogenic drugs. AB - BACKGROUND: Prostate cancer is the most frequent cancer diagnosed in American men today. Currently, about half of all patients with newly diagnosed prostate cancer present with metastatic diseases. METHODS: Antiandrogenic drugs, or more appropriately androgen-receptor antagonists, represent a group of compounds that currently have played a limited role in the treatment of metastatic prostate cancer. Their method of action is primarily one of blocking androgens at their receptor sites in target tissues. They generally are classified as steroidal or nonsteroidal compounds. Cyproterone acetate and megestrol acetate are synthetic steroidal antiandrogenic drugs that, not only compete with testosterone and dihydrotestosterone for androgen receptors, but also have progestational activity and reduce pituitary luteinizing hormone and subsequently plasma testosterone. Nonsteroidal antiandrogenic agents (flutamide, Casodex [ICI Pharmaceuticals, England], and nilutamide) block cellular binding of androgens only, and there is no reduction of testosterone levels. RESULTS: Antiandrogenics have been used in numerous trials both in Europe and the United States. This group of compounds have been used as monotherapy and in combination therapy, ie, with orchiectomy or with LHRH agonists. CONCLUSIONS: Currently, antiandrogens are used primarily in conjunction with conventional medical or surgical castration to achieve maximal androgen deprivation; however, ongoing clinical studies are comparing these compounds alone against standard hormonal therapy. It seems probable that antiandrogens will play an expanding role in the treatment of metastatic prostate cancer as well as having a role in the treatment of prostate cancer. PMID- 8428327 TI - Prostate cancer. Primary hormonal treatment. AB - BACKGROUND: No other treatment matches the 40-60% objective response and up to 80% subjective responses for primary hormonal therapy in patients with symptomatic disease. This makes primary hormonal treatment first-line therapy metastatic prostate cancer. RESULTS: The treatment outcome is determined by the initial and intermittent prognostic factors. In the multihormonal environment, several endocrine treatments have shown competing levels of response. Maximal androgen blockade has emerged as the best treatment to achieve a response and even has increased survival modestly. Monotherapy with nonsteroidal antiandrogens plays a role in patients trying to preserve their potency. CONCLUSIONS: Pilot treatment adjusted to the medical needs and social preferences of the patient is appropriate and allows a tailored approach to the individual patient. PMID- 8428328 TI - Combination therapy for prostate cancer. Endocrine and biologic basis of its choice as new standard first-line therapy. AB - RESULTS. All double-blind randomized and prospective clinical trials in advanced prostate cancer have shown that combination therapy using a nonsteroidal antiandrogen in association with a luteinizing hormone-releasing hormone (LHRH) agonist or orchiectomy has significant benefits according to all the subjective and objective parameters used, the most important being a prolongation of survival ranging from 5.4-15.0 months compared with LHRH agonists or orchiectomy alone (standard therapy). The benefits observed are probably the result of the blockade by the antiandrogenic agents of the androgens of adrenal origin, which represent, on average, 40% of the total androgens in men and which, otherwise, are left free to continue to stimulate prostate cancer after castration. These data are supported well by the demonstration of the expression of the genes encoding all enzymes required for the formation of active androgens in prostatic tissue from the inactive adrenal steroid precursors; this new specialty is called intracrinology. Both fundamental and clinical data indicate that low androgen levels cause changes in the cancer cells that lead to no or a poor response to antihormonal therapy. CONCLUSIONS. It is thus imperative that combination therapy be used as first treatment in all patients in whom endocrine therapy is indicated. Prior exposure to monotherapy shortens the patient's life by many months and produces a poor quality of life. PMID- 8428329 TI - Ketoconazole and liarozole in the treatment of advanced prostatic cancer. AB - BACKGROUND: Ketoconazole, an imidazole derivative, is an orally active antifungal agent. In high doses (400 mg three times a day), it inhibits the biosynthesis of testicular and adrenal androgens and may therefore be useful for the treatment of hormone dependent diseases such as advanced prostatic cancer. Similarly, a new imidazole derivative, liarozole, was recently found to interfere with testicular and adrenal steroid biosynthesis in animals and healthy volunteers. METHODS: The therapeutic and endocrine effects of ketoconazole and liarozole in patients with disseminated prostatic cancer are discussed, including data from the literature and personal experience. RESULTS: Using high-dose ketoconazole, medical castration with the expected clinical response was achieved easily in previously untreated patients in all clinical series (personal data include seven patients). In patients with prostatic cancer who had relapses after castration, few objective remissions were achieved. By contrast, long-lasting subjective remissions, especially pain relief, were seen in more than half of the patients (personal data include 20 patients). Gastrointestinal intolerance, which was the main side effect, severely limits the routine use of the drug. Recently, the authors studied the effect of liarozole on adrenal steroid production in castrated patients whose disease was progressive after first-line treatment. Unlike ketoconazole therapy, adrenal androgen and cortisol levels were not modified. A Phase I-II trial was then done in 44 patients with metastatic prostatic cancer in clinical relapse. In patients with measurable disease, objective responses, including tumoral volume reduction, occurred in approximatively 30%. A prostate specific antigen reduction of 50% or more was noted in approximatively 50% of patients. Pain relief occurred in most patients. Mucocutaneous side effects were observed in most patients--dryness of the skin and onychomalacia. Raised tissue retinoic acid levels suggested a possible pathway by which this drug might exert its cytotoxic effects. CONCLUSIONS: Ketoconazole in high doses is effective in first-line and second-line therapy for advanced prostatic cancer, but gastrointestinal side effects limit its routine use. Liarozole is a new imidazole that is also effective in second-line therapy for prostatic cancer and has fewer side effects. Unlike ketoconazole, its effect is not mediated by inhibition of steroid biosynthesis. PMID- 8428330 TI - Deferred therapy in patients with advanced disease. AB - Disease that is known to have extended beyond the prostatic capsule is unlikely to be cured by radical surgical excision, although the primary tumor may be controlled by external irradiation. The development of progressive (metastatic) disease is common and may be retarded by hormonal therapy. The timing of such treatment and the debate about monotherapy versus total androgen ablation has not been resolved fully. It is considered further in this article. PMID- 8428331 TI - Expanding the definition of quality of life for prostate cancer. AB - BACKGROUND: Quality of life in prostate cancer is a multidimensional concept. From a social work perspective, three quality-of-life issues require additional consideration. These are: the impact of the disease and its treatment on family caregivers, the possibility of age bias and ethical dilemmas in treatment decisions, and the specific concerns of black men in whom the disease is more common. METHODS: Oncology and social work literature was reviewed. RESULTS: Studies of caregivers of elderly patients contribute to understanding the stress and anxieties of families, one of whose members has prostate cancer as a chronic illness. Current debates on expectant management and advanced directives provoke ethical concerns about when to treat prostate cancer especially in elderly patients. A growing appreciation of black health practices and social supports must be considered along with the socioeconomic factors that may inhibit or promote access to care. CONCLUSIONS: Social workers can assist the treatment team in understanding the importance of these factors for specific patients. Additional research is needed on the effect of these issues on access to care, physician-patient-family communication, long-term treatment plans, and outcome. PMID- 8428332 TI - Antiandrogenic agents as monotherapy in advanced prostatic carcinoma. AB - Pure antiandrogens have a quality-of-life advantage over other androgen ablation methods in the treatment of patients with prostatic cancer because they do not reduce the serum testosterone and therefore do not have a marked inhibitory effect on libido and potency. The long half-life of two of the three currently studied pure antiandrogens permits once-a-day administration, which should enhance patient compliance. With continued administration, there is a gradual rise in serum testosterone, and the clinical impact of this requires additional study using randomized Phase III trials. Proper stratification of patients at entry into such studies with documentation of various prognostic factors will add statistical value and enable physicians to draw better conclusion on the relative efficacy of these agents. PMID- 8428333 TI - Endocrine therapy of advanced carcinoma of the prostate. AB - Increasing evidence suggests that growth of the prostatic tissue is regulated by a network of hormones and growth factors, in which androgens play the prominent role. Hormonal manipulation remains the core of treatment for locally advanced and metastatic prostate cancer. Achievement of a complete androgen blockade, by surgical or medical means or a combination of both, offers superior results in palliative management of advanced disease. Management of hormonal refractory cancer, however, remains a challenge to clinicians. PMID- 8428334 TI - High-dose continuous-infusion fosfestrol in hormone-resistant prostate cancer. AB - BACKGROUND: The initial treatment of advanced-stage prostate cancer is total androgen deprivation. Autonomous proliferation of primarily or secondarily hormonal unresponsive cells may explain the development of hormone-refractory status. The median survival of patients with hormone-resistant disease is short; there is no standard regimen of chemotherapy. METHODS: Fosfestrol or diethylstilbestrol diphosphate and its metabolites have cytotoxic activity in hormone-refractory prostatic cell lines. Pharmacokinetic studies have shown that fosfestrol metabolites have a short half-life that supports the use of long-term infusion in the clinic. RESULTS: A review of the literature shows that high-dose fosfestrol induces no objective response, a greater than 50% tumor marker decrease in 50% of patients, a subjective improvement in 75% of patients, and cardiovascular complications in 5% of patients. The median survival time of patients is 5 months after the onset of treatment. CONCLUSIONS: An exact evaluation of the role of high-dose estrogens requires additional investigation. PMID- 8428335 TI - Pain management in the patient with prostate cancer. PMID- 8428336 TI - Quality of life in patients with prostatic cancer. A feasibility study. The Members of Quality of Life Committee of the EORTC Genitourinary Group. AB - BACKGROUND: In recent years, there has been a growing recognition of the need to include parameters representing the patients' view of their conditions that, therefore, are more subjective in nature. METHODS: As a first effort to introduce quality-of-life (QOL) assessment in prostatic cancer clinical trials, the European Organization for Research and Treatment of Cancer (EORTC) Genitourinary Group, in cooperation with the EORTC QOL Group, activated Protocol 30853 (orchiectomy versus goserelin acetate and flutamide in previously untreated patients with Stage M+ disease. Study Coordinator: Louis Denis). The use of patient-administered QOL questionnaires was optional, and of 327 patients, only 22% underwent pretreatment assessments. RESULTS: Psychologic distress, fatigue, social and family life, and pain are the most important to the patient on a subjective basis, and these were confirmed in relation to objective parameters. There was a discrepancy between the doctors' evaluations and the patients' opinions about subjective morbidity, namely, in regard to sexual status and pain. CONCLUSIONS: This EORTC trial revealed the reluctance of clinicians to do QOL research, partly related to feasibility problems and partly to the doctors' doubts about the value of such efforts. QOL assessment should become a mandatory part of clinical trials in prostatic cancer. PMID- 8428337 TI - A comparison of the quality of life of patients with metastatic prostate cancer who received or did not receive hormonal therapy. AB - BACKGROUND AND METHODS: The quality of life in patients who elected to receive hormonal therapy for newly diagnosed asymptomatic metastatic prostate cancer was compared with a comparable group of patients who chose to defer immediate intervention. RESULTS AND CONCLUSIONS: The premise that active therapy, despite its side effects, would improve or maintain the psychosocial quality of life was not substantiated, and the no-therapy group had better physical and sexual functioning. PMID- 8428338 TI - Effect of suramin on the mitogenic response of the human prostate carcinoma cell line PC-3. AB - BACKGROUND: Suramin is an anthelmintic drug that recently has been shown to have clinical efficacy in the treatment of patients with some advanced malignancies, including prostate carcinoma. The current study was done to assess the effect of suramin at clinically relevant doses on the growth in culture of a human prostatic carcinoma cell line, PC-3. METHODS: The antiproliferative effect of varying doses of suramin on PC-3 was assessed. Northern blot analysis was done to assess the potential changes in genetic expression at different times after the initiation of treatment. RESULTS: Suramin inhibited the proliferation of PC-3 in a dose-related manner (concentration range, 30-300 microM). Compared with fetal calf serum 2%, when the cells were grown in fetal calf serum 10%, higher concentrations of suramin were required to inhibit tritiated thymidine incorporation. When grown in RPMI without supplement, the PC-3 cell number remained the same. When 100 microM suramin was included, the cell number decreased. By contrast, when RPMI was supplemented with insulin, transferrin, and selenium (ITS), PC-3 grew well. The inhibition of the proliferation of PC-3 cells by suramin was decreased when ITS were added to the cells grown under serum-free conditions. CONCLUSIONS: These results were consistent with the hypothesis that in vitro inhibition of the growth of PC-3 cells by suramin may be caused, at least in part, by the growth factor antagonism of the drug. In fetal calf serum 2%, the suramin inhibition was reversible after 3 days. If the treatment was extended to 6 days, however, the PC-3 cells were unable to recover. Cell-cycle analysis revealed that, after 6 days of treatment, there was a decrease in the number of cells in G1 that corresponded with an increased number of cells in G2/M. This suggested that critical antineoplastic events were occurring during this time. Molecular analysis did not detect any altered expression of actin, transforming growth factors alpha or beta, or histone compared with untreated control samples. PMID- 8428339 TI - Oncogenes in prostate cancer. An update. AB - Oncogenes have been implicated in the carcinogenic development of many diverse types of human malignancies. For some cancers, the expression of specific oncogenes has been shown to have diagnostic or prognostic value. By contrast currently, no oncogene has been correlated conclusively with the initiation or progression of prostate cancer. The ras oncogene has been investigated the most thoroughly for its involvement in prostate cancer, but ras does not appear to play a significant role in the development of this malignancy. Several years ago, limited studies hinted at the possibility of overexpression of the myc oncogene and aberrant expression of the sis oncogene in prostate cancer, but additional studies to clarify the involvement of these oncogenes have not been done. Oncogenic activity of growth factors or growth factor receptors in prostate cancer has been suggested but not amply demonstrated. Current dogma indicates that oncogenes exist in prostate cancer, but these will be identified only by more intensive investigation. PMID- 8428340 TI - Transgenic models for the study of prostate cancer. AB - Transgenic model systems provide tools for obtaining information that clarifies important relationships between genetic alterations and carcinogenesis. One such relationship is the induction of specific growth factor activities by dominantly acting oncogenes. Using a "transgenic organ" model referred to as mouse prostate reconstitution (MPR) under conditions where the ras and myc oncogenes were introduced using a recombinant retrovirus into both the mesenchymal and epithelial compartments of the urogenital sinus, poorly differentiated prostate cancer (PC) was produced with high frequency (> 90%) in inbred C57BL/6 mice. Time course studies using northern blotting and immunohistochemical analysis showed that the transition from benign to malignant status invariably was associated with the induction of elevated transforming growth factor-beta 1 (TGF-beta 1) expression. Additional immunohistochemical analysis of TGF-beta 1 in human PC and benign prostatic hyperplasia (BPH) showed that positive extracellular staining was significantly more extensive in PC compared with BPH. This differential staining pattern was evident in focal areas of PC adjacent to BPH. These findings in both the MPR model system and human PC suggest that elevated TGF-beta 1 expression is involved in the progression to malignancy and that its pattern of expression may become a useful marker of PC. Additional studies using transgenic animal models will continue to provide important clinically useful information about PC in man. PMID- 8428341 TI - Recessive oncogenes. AB - Tumor-suppressor genes (antioncogenes or recessive oncogenes) are cancer genes that achieve their oncogenic effect by mutational inactivation of both normal alleles. By contrast, oncogenes are created from protooncogenes by mutations that lead to aberrant functional activation. Mutation of multiple suppressor genes and/or oncogenes probably is required for the genesis of most human neoplasms. Two well-characterized tumor-suppressor genes, the retinoblastoma gene (rb) on chromosome 13q and p53 on chromosome 17p, frequently are mutated in a broad range of human cancer types. Mutations of these genes have been documented in prostate carcinoma but appear to affect only a subset of cases. Nevertheless, as in other cancers, introduction of normal copies of rb or p53 suppresses the neoplastic properties of prostatic tumor cells carrying mutated alleles of the relevant gene. These results suggest that mutation of rb or p53 is involved in the genesis or progression of some prostate cancers. Frequent allelic losses of certain chromosome arms (especially 8p, 10p and q, and 16q) from prostatic cancer cells may indicate the involvement of novel suppressor genes located in these regions. Although the inactivation of suppressor genes appears to be a common genetic mechanism in human oncogenesis, the rates of mutation of particular genes vary widely with the type of cancer. It is unknown whether prostate cancers with or without mutation of rb, p53, or other suppressor loci differ biologically or prognostically; this is an area of active investigation. Fundamental understanding of the genetic lesions that occur during human oncogenesis has great potential for clinical application in diagnosis, prognosis, and therapy. PMID- 8428342 TI - Prostate cancer. An overview of an increasing dilemma. PMID- 8428343 TI - Prostate cancer. Magnitude of the problem. American College of Surgeons database and epidemiologic study. AB - BACKGROUND: The third American College of Surgeons pattern of care survey for cancer of the prostate is underway to document the current methods and means of presentation, diagnosis, and clinical management of patients in the United States. METHODS: Questions about the geographic distribution and methods of treatment by stage and grade together with patient mix and survival information will be answered from the data collected from more than 950 hospitals across the United States for the period 1984-1990. May 1992 was the final date for submission of data for this survey. CONCLUSIONS: Prostate cancer continues to be the most common cancer in men, excluding skin cancer. The annual number of patients with this disease reported in the National Cancer Database for 1988 was 26,957, an increase of 9427 cases yearly since the end of the second American College of Surgeons' survey in 1983. PMID- 8428344 TI - Prostate cancer. Radiation therapy for localized disease. AB - BACKGROUND: Since 1910, a variety of radiation modalities, including radioactive isotopes, photons, and particle beams, have been used to treat prostatic cancer. METHODS: This report focuses on external beam x-irradiation produced by medium energy linear accelerators. Between 1956 and 1990, 1119 patients have been treated with curative intent at Stanford University. Tumor doses of 70 Gy delivered at 10 Gy/wk have been safe and effective. RESULTS: Fifteen-year survival rates ranging from 50%, equivalent to that of an age-matched cohort, for the least extensively localized tumors to 18% for the most extensive have been achieved. Survival is inversely proportional to clinical stage and histopathologic grade. CONCLUSION: Although external beam radiation therapy has been found to be safe and effective for the treatment of prostatic cancer, improvement in results of treatment of the more advanced tumors might be achieved by combining external beam and interstitial irradiation. This would achieve a higher radiation dose within the tumor. Alternatively, the treatment can be augmented with hyperthermia or other sensitizers in order to achieve a higher biological dose. PMID- 8428345 TI - Prostate brachytherapy. An overview. AB - BACKGROUND: Prostate brachytherapy represents one of the oldest techniques of using radiation therapy to treat prostate cancer. Over the past 10 years, there have been major changes in the types of prostate brachytherapy that can be performed with the introduction of new radioactive isotopes, new afterloading techniques, and an improved understanding of the radiobiology associated with differing dose rates. METHODS: Prostate brachytherapy can be divided into temporary implantation using high activity sources such as iridium-192, or permanent brachytherapy using the interstitial implantation of iodine-125 or palladium-103 sources. There are various techniques that can be used to actually insert the radioactive material into the prostate. This can be done as an open or closed procedure and can be performed via a suprapubic or a perineal retropubic approach. The use of remote afterloading has substantially reduced the radiation protection problems associated with manually loaded radioactive sources. RESULTS: Results using brachytherapeutic isotopes in the treatment of prostate cancer have been variable, but it appears that using the higher dose rate sources and delivering a high relative integral dose to the prostate can result in improved histologic control of prostate cancer. In patients with aggressive prostate cancer, the use of iodine-125 permanent implantation has not been successful. CONCLUSIONS: The role of brachytherapy in the treatment of prostate cancer remains an exciting alternative in the management of prostate cancer. Its role is becoming more defined in the treatment of large, bulky prostatic neoplasms as a way of improving the dose distribution achieved between normal and tumor tissue. PMID- 8428346 TI - Conservative approaches to the management of localized prostatic cancer. AB - BACKGROUND: The management of early stage prostatic cancer is controversial. METHODS: Pertinent literature concerning the conservative management of early stage prostatic cancer by early endocrine therapy (EET) or by deferred treatment (DT) was reviewed. RESULTS: EET has not been systematically studied. Available evidence suggests that early stage prostatic cancer often progresses slowly and that DT results in a cancer-specific mortality of approximately 80% at 10 years. CONCLUSIONS: EET warrants clinical investigation. DT is a management option, at least in patients with a life expectancy of 10 years or less. PMID- 8428347 TI - Salvage surgery for radiation failure in prostate cancer. AB - METHODS: Salvage surgery was done in 43 patients who did not respond to radiation therapy of prostate cancer between 1982-1991. Thirty-five patients underwent salvage prostatectomy and 8, cystoprostatectomy. RESULTS: The complications were significant; four patients had rectal injuries (all closed primarily), one had a ureteral injury, and there was one perioperative death. Urinary incontinence occurred in 10 of 35 patients (30%). Pathologic step sections of the prostate showed that only 13 of 43 patients (30%) had negative surgical margins. Follow-up (range, 1-10 years) revealed that 34 patients were alive, and 9 had died. Eleven of 20 patients were alive who were followed more than 5 years. Ten patients were considered to have no evidence of disease (undetectable prostate specific antigen levels). CONCLUSION: In selected patients, salvage surgery has a place in the treatment of prostate cancer after radiation therapy failure. PMID- 8428348 TI - Screening and early detection of prostate cancer. AB - BACKGROUND: The value of routine screening for prostate cancer remains unknown because of the lack of randomized studies. Until such studies are completed, a sensible approach to screening is needed. METHODS: One approach is to use the method of decision analysis to model the outcome of alternative screening strategies. CONCLUSIONS: By assessing the results in terms of their effect on life expectancy and prostate cancer mortality, it may be possible to select the policy that has the greatest potential benefit for the least cost. PMID- 8428349 TI - Prostatic microcarcinomas in relation to cancer origin and the evolution to clinical cancer. AB - BACKGROUND: In the prostate there is a uniquely high prevalence of microcarcinomas relative to larger cancers. The probability of metastasis has been found to correlate closely with increasing cancer volume. It is important to explore the relationship of microcarcinomas to these larger clinically significant cancers by comparing their histologic features and to trace their relationship to dysplasia, a proposed precursor lesion. METHODS: The histologic features of 107 microcarcinomas were related to associated dysplasia lesions. Their histologic grade was quantitated and compared with the quantitative grade and volume distribution of 100 incidental autopsy cancers and 209 clinical cancers from radical prostatectomy. RESULTS: Microcarcinomas were contiguous with dysplasia foci in 81% of the cases, and evidence of cancer origin was found in 48%. Only 3 of 107 microcarcinomas had Grade 4 (poorly differentiated) areas. Grade 4 frequency and extent increased with cancer volume, and the relationship was similar between the autopsy and prostatectomy series. Metastasis was found only with cancers larger than 4 cc and having Grade 4 areas. CONCLUSIONS: The majority of prostatic microcarcinomas appeared to arise from dysplasia. Only 3% of these early cancers showed areas of high histologic grade. In the autopsy and clinical series, it appeared that poorly differentiated areas evolve from low grade cancer with time and increasing volume. Probability of metastasis was a function of volume and grade. PMID- 8428350 TI - Adjuvant therapy in prostatic cancer. AB - BACKGROUND: Prostatic cancer is an unusual malignancy because, commonly, it is slowly progressive and occurs at an age where the patient's life expectancy is short as a result of considerable competing mortality rates from other diseases. In addition, the place of radical surgery, i.e., radical prostatectomy is still not defined completely or established uniformly. METHODS: This article attempts to offer comparisons of the initial forms of primary therapy and to determine the way in which adjuvant therapy may be tested and its results assessed. RESULTS AND CONCLUSION: Additional information should come from large-scale randomized clinical trials involving several hundred patients in each treatment arm. PMID- 8428351 TI - Increased generation of lipid-derived and ascorbate free radicals by L1210 cells exposed to the ether lipid edelfosine. AB - Using the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone, we have detected a lipid-derived carbon-centered free radical generated from intact L1210 lymphoblastic leukemia cells that were exposed to 1-O-octadecyl-2-O-methyl-rac glycero-3-phosphocholine (edelfosine or ET-18-OCH3) and oxidative stress. The spectral characteristics, including hyperfine splitting constants of aN = 15.61G and aH = 2.65G, were consistent with the spin trapping of an alkyl radical. Radical detection required iron and prior enrichment of cellular components with the polyunsaturated fatty acid docosahexaenoic acid; unmodified cells failed to generate detectable free radical. Ascorbate further enhanced radical generation. The detection of lipid-derived free radicals when intact cells are exposed to edelfosine provides further evidence that oxidative stress may play an important role in the cytotoxic mechanism of this class of anticancer drug. PMID- 8428352 TI - Metabolites of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone in smokers' urine. AB - Metabolites of the tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3 pyridyl)-1-butanone, a potent pulmonary carcinogen, have been quantified in the urine of 11 smokers. They were not detected in nonsmokers' urine. The metabolites, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol and its glucuronide, were detected in quantities of 0.23-1.0 and 0.57-6.5 micrograms/24 h, respectively. The results of this study provide the first evidence for metabolites of tobacco-specific nitrosamines in human urine. PMID- 8428353 TI - Plasma and cerebrospinal fluid pharmacokinetic study of topotecan in nonhuman primates. AB - Topotecan, a water soluble semisynthetic analogue of camptothecin, is a topoisomerase I inhibitor that has recently entered phase II clinical trials. Topotecan has shown significant preclinical activity in refractory murine tumors and in human tumor xenograft models. In addition, objective antineoplastic activity has been observed in recent adult phase I clinical trials. Topotecan is unstable in solution and is rapidly and spontaneously converted to a less active open ring form which predominates at physiological pH. This study was undertaken to better define the pharmacokinetic behavior of this highly unstable compound in both plasma and cerebrospinal fluid (CSF) and to measure the degree of CSF penetration of this novel antineoplastic agent. Three nonhuman primates with indwelling Ommaya reservoirs received 10 mg/m2 i.v. topotecan administered as a 10-min infusion. Frequent plasma and CSF samples were obtained and immediately extracted and assayed with a reverse phase high performance liquid chromatography assay to quantitate the concentration of topotecan (lactone). Samples were then acidified and reinjected to quantitate total drug (lactone ring plus open ring). Peak plasma concentrations of topotecan ranged from 0.27 to 0.45 microM. Plasma disappearance of the lactone ring was biexponential with a distribution half-life (t1/2 alpha) of 22 +/- 5 min and an elimination half-life (t1/2 beta) of 1.3 +/- 0.1 h. Total body clearance of topotecan was 72.1 +/- 15.8 liters/h/m2. The volume of distribution at steady state was 88.6 +/- 33.2 liters/m2. Peak CSF concentrations of topotecan occurred at 30 min following drug administration and ranged from 0.044 to 0.074 microM. CSF disappearance paralleled that in plasma. The mean ratio of the area under the CSF concentration-time curve to that in plasma was 0.32 (range, 0.29 to 0.37). The mean CSF penetration of topotecan exceeds 30%, which is significantly greater than the penetration of most structurally similar chemotherapeutic agents. The impact of chemotherapy on the survival of patients with primary or metastatic central nervous system malignancies is very limited. Therefore, this novel antineoplastic agent is an excellent candidate for further study in patients with high risk or refractory central nervous system tumors. PMID- 8428354 TI - Neutralizing antibodies to human interleukin 6 reverse hypercalcemia associated with a human squamous carcinoma. AB - Bone resorbing cytokines may be associated with abnormalities in bone cell function and calcium homeostasis in a number of pathological conditions. One of these cytokines is interleukin 6 (IL-6), which is a multifunctional cytokine which has been shown to be associated with increased formation of bone resorbing osteoclasts in vitro. In this report, we demonstrate that neutralizing antibodies to human IL-6 lower the blood calcium in nude mice carrying a human tumor associated with increased IL-6 production. There was no effect on blood calcium in hypercalcemic nude mice carrying tumors not associated with increased IL-6 production and normocalcemic tumor-bearing nude mice. These results suggest that increased production of IL-6 may contribute to disturbances in calcium homeostasis in some malignancies, and suggest that neutralization of IL-6 effects can lower the serum calcium in these tumors. PMID- 8428355 TI - Distinct infrared spectroscopic patterns of human basal cell carcinoma of the skin. AB - Infrared spectroscopy combined with high pressure (pressure-tuning infrared spectroscopy) was applied to the study of paired sections of basal cell carcinomas (BCC) and normal skin from ten patients. Atmospheric pressure IR spectra from BCC were dramatically different from those from the corresponding normal skin. Compared to their normal controls, BCCs displayed increased hydrogen bonding of the phosphodiester group of nucleic acids, decreased hydrogen bonding of the C--OH groups of proteins, increased intensity of the band at 972 cm-1, a decreased intensity ratio between the CH3 stretching and CH2 stretching bands, and accumulation of unidentified carbohydrates. Some of these changes are shared by all human epithelial malignancies studied to date, while some others appear as yet unique to basal cell carcinoma. The diagnostic value of infrared spectroscopy in BCC remains to be determined. PMID- 8428356 TI - Alkylamides as inducers of human leukemia cell differentiation: a quantitative structure-activity relationship study using comparative molecular field analysis. AB - Computer assisted quantitative structure-activity studies using comparative molecular field analysis (CoMFA) were performed on a series of alkylamides that induce cell differentiation. The series included alkylformamides, alkylacetamides, alkylureas, and substituted hexyl analogues of acetamide. The biological activity studied for correlation with structure was the ability of each compound to induce differentiation of the human promyelocytic leukemia cell line, HL-60, to granulocyte-like cells. In the CoMFA study, both steric and electrostatic fields were used along with molecular weight to determine a correlation between biological activity of the compounds and their structural features. The CoMFA results indicated a linear structure-activity correlation with a high predictive value. There was almost an even contribution towards activity from steric interactions, electrostatic potential, and molecular weight. These findings confirm a previous report by Langdon and Hickman (S. P. Langdon and J. A. Hickman, Cancer Res., 47: 140-144, 1987) that the ability to induce cell differentiation is highly dependent on molecular weight. Additionally, CoMFA contour maps provided information about regions of the molecule that are favorable to increased steric bulk and electrostatic charge. CoMFA was used to predict the activities of six hexamethylene acetamide analogues: ethyl 6 acetamidohexanoate; 6-acetamidohexanol; 1,5-bis(acetamido)hexane; 6 acetamidohexanonitrile; 6-acetamidohexanoic acid; and caprolactam. Although the model incorrectly predicted high activity for 6-acetamidohexanoic acid, the predicted activities for the remaining compounds were 0.3 to 1.5 times that of the corresponding experimental activities, which is comparable to the results obtained from other published CoMFA studies. PMID- 8428357 TI - Chemoprevention of oral carcinogenesis by DL-alpha-difluoromethylornithine, an ornithine decarboxylase inhibitor: dose-dependent reduction in 4-nitroquinoline 1 oxide-induced tongue neoplasms in rats. AB - The modifying effect of three doses of DL-alpha-difluoromethylornithine (DFMO) given p.o. during the post-initiation phase of tongue carcinogenesis initiated by 4-nitroquinoline 1-oxide (4-NQO) was studied in male ACI/N rats. Animals were given 4-NQO at 20 ppm for 8 weeks in the drinking water to induce tongue neoplasms. One week after the stop of 4-NQO treatment, rats were transferred to the drinking water containing DFMO at concentrations of 100, 1000, and 2000 ppm for 25 weeks. The other groups consisted of rats given 2000 ppm DFMO alone or untreated rats. Thirty-four weeks after the start of the experiment, all animals were necropsied, and the incidences of neoplasms and preneoplastic lesions in the tongue, polyamine levels in the bloods and tongue tissues, and cell proliferation estimated by the number and area of silver-stained nucleolar organizer regions in the tongue epithelium were compared among the groups. Feeding of DFMO at all doses significantly inhibited the incidence of tongue neoplasms compared to the group given 4-NQO alone. DFMO at levels of 1000 and 2000 ppm significantly reduced the incidence of preneoplastic lesions of the tongue. Results analyzed by the linear regression method suggested a dose-dependent inhibition in the incidences of neoplastic and preneoplastic lesions of the tongue with increasing levels of DFMO. Increased levels in polyamines in the blood and tongue tissue were significantly suppressed by the treatment of DFMO. Also, silver-stained nucleolar organizer region indices were significantly reduced by the DFMO exposure. These results indicate that increasing levels of DFMO in the drinking water inhibited 4-NQO-induced tongue carcinogenesis in a dose-dependent manner and such inhibition was related to reduction in the polyamine levels of blood and tissue and decrease in the cell proliferation. PMID- 8428358 TI - Two types of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone hemoglobin adducts, from metabolites which migrate into or are formed in red blood cells. AB - The tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone (NNK), is considered to play an important role in the induction of lung cancer in tobacco users. In rats treated with [5-3H]NNK, 20 to 40% of the tritium bound to hemoglobin (Hb) is released by base hydrolysis as 4-hydroxy-1-(3-pyridyl)-1 butanone (HPB). This HPB-releasing adduct has been quantified in tobacco users and is considered a biochemical marker for uptake and activation of tobacco specific nitrosamines. In this paper we report the formation of this adduct in red blood cells (RBC) cultured for 2 h with hepatocytes and 5 microM NNK (6.35 +/ 0.21 fmol HPB/mg Hb). The HPB-releasing adduct was not formed in RBC in the absence of hepatocytes (< 0.5 fmol/mg Hb). Therefore, the HPB-releasing adduct must form from a pyridyloxobutylating metabolite of NNK which traveled out of the hepatocytes and into RBC where it reacted with Hb. Other distinct Hb adducts were formed when NNK was incubated with RBC alone. 4-Oxo-4-(3-pyridyl)butyric acid was detected by radio flow high-performance liquid chromatography in the media of these incubations. The Hb isolated from RBC incubated with [C3H3]NNK contained as much as 10 times more covalently bound tritium than the Hb from [5-3H]NNK-treated cells. [C3H3]-1-Methylhistidine and [C3H3]-S- methyl-cysteine were formed when [C3H3]NNK was incubated with the 25,000 x g supernatant from RBC. This supernatant contains 50 mg Hb/ml. We propose that Hb mediates alpha hydroxylation of NNK at the methylene carbon. The alpha-hydroxynitrosamine formed decomposes to methanediazohydroxide and 4-oxo-1-(3-pyridyl)butanal. The former would methylate nucleophilic sites in Hb, i.e., cysteine and histidine. The latter would bind to Hb or be further oxidized to 4-oxo-4-(3-pyridyl)butyric acid. The ability of the RBC to activate NNK to Hb-binding species stresses the importance of understanding how a particular adduct is formed prior to its use as a biochemical marker or internal dose monitor. PMID- 8428359 TI - The antiproliferative effect of dietary calcium on colonic epithelium is mediated by luminal surfactants and dependent on the type of dietary fat. AB - Bile acids and fatty acids may promote colon cancer by inducing colonic hyperproliferation. Dietary calcium inhibits the promoting effects of bile acids and fatty acids, possibly by precipitating these surfactants and lowering their cytolytic activity. Because bile acids and fatty acids are products of fat digestion, their effects may be dependent on the type of dietary fat. The effects of the type of dietary fat (energy percentage, 40) and of CaHPO4 supplementation (25 versus 225 mumol/g diet) on the luminal solubility of surfactants, cytolytic activity, epitheliolysis, and in vivo colonic proliferation were studied in rats using Western high-risk diets. The different types of commercially available fats were butter, saturated margarine, and polyunsaturated margarine. Supplemental calcium drastically increased fecal fatty acid excretion, the effect being dependent on the type of fat, and slightly stimulated fecal bile acid excretion. Soluble surfactant concentrations were drastically decreased by calcium supplementation with all three types of dietary fat. Consequently, cytolytic activity of fecal water was decreased by supplemental calcium. These luminal effects of calcium resulted in a lower intestinal epitheliolysis. The compensatory proliferation of the colonic epithelium was decreased by supplemental CaHPO4 for the butter and saturated margarine diets. Despite CaHPO4 dependent decreases in luminal effects and epitheliolysis, no significant decrease in proliferation on the polyunsaturated margarine diet was observed. Multiple regression analysis of soluble surfactants with cytolytic activity (R = 0.76), epitheliolysis (R = 0.74), and colonic proliferation (R = 0.84) showed highly significant associations. Cytolytic activity and epitheliolysis as well as epitheliolysis and proliferation were highly correlated (r = 0.97 and r = 0.88, respectively; n = 36) for control and CaHPO4-supplemented diets, suggesting cause and-effect relationships. It is concluded that the antiproliferative effect of dietary calcium is mediated by the precipitation of luminal surfactants and is dependent on the type of dietary fat. PMID- 8428360 TI - Serum micronutrients and the subsequent risk of oral and pharyngeal cancer. AB - To investigate the relationship between serum micronutrients and the subsequent risk of oral and pharyngeal cancer, a nested case-control study was conducted within a cohort of 25,802 adults in Washington County, MD, whose blood samples were collected in 1974 and stored at -70 degrees C for subsequent assays. The serum levels of nutrients in 28 individuals who developed oral and pharyngeal cancer during 1975 to 1990 were compared with levels in 112 matched controls. Serum levels of all individual carotenoids, particularly beta-carotene, were lower among subjects who developed oral and pharyngeal cancer. The risks of this malignancy decreased substantially with increasing serum level of each individual carotenoid. Persons in the highest tertile of total carotenoids had about one third the cancer risk as those in the lowest tertile. High serum levels of alpha tocopherol also were related to a low oral cancer risk in later years, but the risks were elevated significantly with increasing serum levels of gamma tocopherol and selenium. The findings from this study are consistent with many previous epidemiological investigations of dietary factors for oral and pharyngeal cancer and provide further evidence for the potential role of carotenoids and alpha-tocopherol in the chemoprevention of these malignancies. PMID- 8428361 TI - Role of carrier ligand in platinum resistance of human carcinoma cell lines. AB - We have examined the effects of the cis-diammine and 1,2-diaminocyclohexane (dach) carrier ligands on cytotoxicity, platinum accumulation and efflux, platinum incorporation into DNA, cytotoxicity of Pt-DNA adducts, and repair of Pt DNA adducts in the human ovarian carcinoma A2780 cell line, the human colon carcinoma HCT8 cell line, and their cis-diamminedichloroplatinum(II) (cisplatin) resistant derivatives, A2780/DDP and HCT8/DDP. The A2780/DDP cell line was 7.7 fold resistant to cisplatin, and the HCT8/DDP cell line was 1.6-fold resistant to cisplatin compared to their parental cell lines. Both were considered as examples of acquired cisplatin resistance. The HCT8/S cell line was 4.6-fold resistant to cisplatin compared with the A2780/S cell line and was considered an example of intrinsic resistance. Decreased accumulation of cisplatin made a significant contribution to acquired cisplatin resistance in the A2780/DDP cell line, probably contributed to intrinsic resistance in the HCT8/S cell line, but made little or no contribution to acquired resistance in the HCT8/DDP cell line. Decreased cytotoxicity of Pt-DNA adducts made a major contribution to both acquired and intrinsic cisplatin resistance in all three cell lines. Increased repair activity made a significant contribution to the decreased cytotoxicity of Pt-DNA adducts in the HCT8/S cell line, a weak contribution in the A2780/DDP cell line, and no contribution in the HCT8/DDP cell line. Glutathione levels were elevated in all the cell lines with acquired and intrinsic resistance, but the increased glutathione levels were not associated with decreased incorporation of platinum into DNA. These data suggest that both decreased accumulation and increased repair contribute to cisplatin resistance to different degrees in these human carcinoma cell lines. In addition, mechanism(s) other than repair may contribute to the decreased cytotoxicity of cis-diammine-Pt-DNA adducts. Of the cells with acquired cisplatin resistance, the HCT8/DDP cell line showed no resistance to tetrachloro(trans-DL)1,2-diaminocyclohexaneplatinum(IV) (ormaplatin, formerly known as tetraplatin), while the A2780/DDP cell line was just as resistant to ormaplatin as to cisplatin. The intrinsically cisplatin resistant HCT8/S cell line showed only partial cross-resistance to ormaplatin. The effects of the dach carrier ligand on both acquired and intrinsic resistance in these cell lines appeared to occur primarily at the level of cytotoxicity of dach-Pt adducts, but the differences in the cytotoxicity of cis-diammine-Pt and dach-Pt adducts could not be explained by differences in repair of those adducts.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8428362 TI - Biochemical and cellular pharmacology of 1843U89, a novel benzoquinazoline inhibitor of thymidylate synthase. AB - Studies on a series of benzoquinazoline folate analogues as inhibitors of human thymidylate synthase led to the selection of 1843U89 for further evaluation. This compound had a Ki of 90 pM versus human thymidylate synthase and was noncompetitive with (6R,S)-5,10-methylenetetrahydrofolate. It was a good substrate for the addition of the second glutamate by hog liver folylpolyglutamate synthetase, having a Vmax/Km value 7.8-fold higher than (6R,S) tetrahydrofolate. The data indicate that 1843U89 was transported into cells via the reduced folate carrier. The Kt for 1843U89 in MOLT-4 cells was 0.33 microM, which was 3-fold lower than that for methotrexate and 16-fold lower than that for (6S-5-formyltetrahydrofolate. V/K values were 20.3 for 1843U89 versus 1.2 and 1.9 for methotrexate and (6S)-5-formyltetrahydrofolate, respectively. It was a potent inhibitor of the growth of human cells, having 50% inhibitory concentrations below 1 nM for all cell lines tested. Growth inhibition was reversed by thymidine alone, indicating that thymidylate synthase was the only site of action of this compound. Growth inhibition was not affected by (6R,S-5-formyltetrahydrofolate at concentrations below 5 microM. However, the 50% inhibitory concentration increased when the concentration in the medium was increased to 100 microM, presumably due to competition for transport. Relative to the human cell lines used, murine cell lines were 80-1300-fold less sensitive to 1843U89 and the other benzoquinazolines tested. This decreased sensitivity appeared to be due, at least in part, to decreased transport or accumulation in murine cells. Ki values for inhibition of methotrexate transport for the benzoquinazolines were 5-17-fold higher in L1210 cells than in MOLT-4 cells. 1843U89, the benzoquinazoline which was transported most efficiently and which was the most potent inhibitor of the growth of human cells, exhibited the largest difference between binding to the MOLT-4 human and L1210 murine transporter. The V/K for L1210 transport was 80 fold less than that for MOLT-4. Initial antitumor studies, using the human thymidine kinase-deficient line GC3TK- to circumvent problems associated with murine transport as well as the high circulating thymidine levels in mice, indicated that 1843U89 had marked in vivo antitumor activity. PMID- 8428363 TI - Pseudomonas exotoxin-based immunotoxins containing the antibody LL2 or LL2-Fab' induce regression of subcutaneous human B-cell lymphoma in mice. AB - We have produced immunotoxins using LL2, a monoclonal antibody which binds to human B-cell lymphomas and which, in a radioiodinated form, induced responses in lymphoma patients (D.M. Goldberg et al., J. Clin. Oncol., 9: 548-564, 1991). We have coupled LL2 to Lys-PE38KDEL, a derivative of Pseudomonas exotoxin (PE) which does not bind to the PE receptor. LL2-PE38KDEL was cytotoxic toward several Burkitt's lymphoma lines, with 50% inhibitory concentration values ranging from 2 to 6 ng/ml (10-30 pM). Another immunotoxin, LL2-Fab'-PE38KDEL, was produced by chemically coupling the Fab' fragment of LL2 to Lys-PE38KDEL. LL2-Fab'-PE38KDEL also was cytotoxic toward the Burkitt's cells, with a 50% inhibitory concentration of 1-2 ng/ml (13-24 PM). The antibody LL2 alone had no cytotoxicity toward the malignant cells, and excess LL2 prevented the cytotoxicity of LL2 PE38KDEL and LL2-Fab'-PE38KDEL. Control immunotoxins UPC-10-PE38KDEL and Mu-9 Fab'-PE38KDEL were not cytotoxic. LL2-PE38KDEL and LL2-Fab'-PE38KDEL bound to cells with 50% and 17% of the affinity of LL2, respectively. Both immunotoxins, but not UPC-10-PE38KDEL, prevented the development of CA-46 tumors in nude mice. LL2-PE38KDEL and LL2-Fab'-PE38KDEL, but not the control immunotoxins, led to complete regressions of measurable s.c. CA-46 tumors in nude mice, when given at 50% and 35% of the 50% lethal dose, respectively. LL2 alone significantly retarded the growth of CA-46 tumors but did not cause complete tumor regressions. Immunotoxins containing derivatives of Pseudomonas exotoxin can be targeted to human B-cell lymphoma and merit further study as potential therapeutic agents. PMID- 8428364 TI - gamma-Interferon plays a key role in T-cell-induced tumor regression. AB - Recent studies have demonstrated that noncytolytic T-cells can mediate regression of murine tumors. In this report, we demonstrate that MCA-105 tumor-draining lymph node cells (DLN) activated with the protein kinase C activator, bryostatin 1, plus a calcium ionophore are capable of inducing specific tumor regression in vivo when adoptively transferred to mice with established metastases. However, these activated DLN cells lack in vitro cytotoxicity against autologous tumor. Antibody against gamma-interferon (IFN-gamma) markedly inhibited the therapeutic efficacy of these activated DLN cells. Anti-tumor necrosis factor produced a statistically significant but weaker inhibition of tumor regression. IFN-gamma, but not tumor necrosis factor alpha, could be shown to be secreted by activated DLN cells in vitro in response to specific tumor. Secretion of IFN-gamma was primarily a function of CD8+ T-cells. IFN-gamma was not directly cytotoxic to sarcoma cells in vitro. Moreover, tumor cells incubated with IFN-gamma were not more susceptible to lysis by activated DLN cells. However, recombinant murine IFN gamma had a significant antiproliferative effect against MCA-105 tumor cells when tested in a [3H]thymidine uptake assay. Similarly, supernatants obtained from DLN/autologous tumor cocultures markedly inhibited MCA-105 proliferation; this antiproliferative effect was abrogated by the addition of anti-IFN-gamma antibody to the cultures. These results suggest that secretion of IFN-gamma by adoptively transferred DLN cells plays an essential role in tumor rejection. The dominant effect of IFN-gamma may be its demonstrated antiproliferative activity. PMID- 8428365 TI - Reshaping a human antibody to inhibit the interleukin 6-dependent tumor cell growth. AB - The mouse PM-1 monoclonal antibody binds to the human interleukin 6 receptor, inhibits IL-6 functions, and shows strong antitumor cell activity against multiple myeloma cells. In order to be effective as a therapeutic agent administered to human patients in repeated doses, reshaped human PM-1 antibodies consisting of human REI-based light chain and NEW-based heavy chain variable regions were designed and constructed with the assistance of a structural model of the mouse PM-1 variable regions. The best reshaped human PM-1 antibody is equivalent to mouse or chimeric PM-1 antibody in terms of antigen binding and growth inhibition against multiple myeloma cells. Only a few minor changes in the human framework regions were required to recreate the mouse PM-1 antigen-binding site within a human antibody. The reshaped human PM-1 antibody, therefore, could be efficacious in human multiple myeloma patients. PMID- 8428366 TI - von Hippel-Lindau syndrome: cloning and identification of the plasma membrane Ca(++)-transporting ATPase isoform 2 gene that resides in the von Hippel-Lindau gene region. AB - We have isolated and analyzed full-length complementary DNA clones encoded by a 200-kilobase gene encompassing the D3S601 locus that resides in the von Hippel Lindau (VHL) gene region. The deduced amino acid sequence shows 99% identity with the published sequence of the rat plasma membrane Ca(++)-transporting ATPase isoform 2 complementary DNA, implying that we have cloned and positioned the human plasma membrane Ca(++)-transporting ATPase isoform 2 gene within the VHL critical region. The gene is expressed in VHL target tissues and should be considered a potential candidate gene for the VHL disease. PMID- 8428367 TI - An antisense oligodeoxynucleotide that depletes RI alpha subunit of cyclic AMP dependent protein kinase induces growth inhibition in human cancer cells. AB - Enhanced expression of the RI alpha subunit of cyclic AMP-dependent protein kinase type I has been correlated with cancer cell growth. We provide evidence that RI alpha is a growth-inducing protein that may be essential for neoplastic cell growth. Human colon, breast, and gastric carcinoma and neuroblastoma cell lines exposed to a 21-mer human RI alpha antisense phosphorothioate oligodeoxynucleotide (S-oligodeoxynucleotide) exhibited growth inhibition with no sign of cytotoxicity. Mismatched sequence (random) S-oligodeoxynucleotides of the same length exhibited no effect. The growth inhibitory effect of RI alpha antisense oligomer correlated with a decrease in the RI alpha mRNA and protein levels and with an increase in RII beta (the regulatory subunit of protein kinase type II) expression. The growth inhibition was abolished, however, when cells were exposed simultaneously to both RI alpha and RII beta antisense S oligodeoxynucleotides. The RII beta antisense S-oligodeoxynucleotide alone, exhibiting suppression of RII beta along with enhancement of RI alpha expression, led to slight stimulation of cell growth. These results demonstrate that two isoforms of cyclic AMP receptor proteins, RI alpha and RII beta, are reciprocally related in the growth control of cancer cells and that the RI alpha antisense oligodeoxynucleotide, which efficiently depletes the growth stimulatory RI alpha, is a powerful biological tool toward suppression of malignancy. PMID- 8428368 TI - The role of cathepsin D in the invasiveness of human breast cancer cells. AB - The aspartyl protease cathepsin D has been shown to be a marker of poor prognosis when found at high levels in primary breast tumors. It has been suggested that this is because the production of cathepsin D increases the invasive potential of the tumor cells, thus increasing the probability of metastasis. We have therefore conducted experiments to determine if secreted cathepsin D makes a significant contribution to the invasive phenotype of breast cancer cells in the Boyden chamber assay of invasion, which measures the ability of a cell to invade through an artificial basement membrane. Cathepsin D secretion and Boyden chamber invasiveness were measured in nine clones of the breast cancer cell line MCF-7, and no correlation was found between cathepsin secretion and invasive behavior. Invasion assays were also conducted in the presence of the aspartyl protease inhibitor pepstatin A, and no inhibition of the invasive behavior of cells was seen. Since low-pH environments are required for both the activation of pro cathepsin D and the activity of the mature enzyme, assays were also conducted in the presence of chloroquine to neutralize the pH in the acidic compartments of the cells. This treatment did not inhibit invasiveness. Cathepsin D secretion by the breast cancer cell lines MDA-MB-231, MDA-MB-435, MDA-MB-435s, MDA-MB-468, SK Br-3, and MCF-7-ADRr was also measured. Again, there was no correlation with invasion. In fact, cathepsin D levels were inversely correlated with aggressive behavior in vivo and in vitro in previously reported studies. These data suggest that cathepsin D secretion by tumor cells is not an important determinant of the invasiveness of the tumor cells per se. These data also reinforce the view that the poor prognosis in clinical breast cancer linked to high tumor levels of cathepsin D is probably due to high levels of cathepsin D in the stromal components of the tumor such as infiltrating inflammatory cells. PMID- 8428369 TI - Studying clonal heterogeneity in human cancers. PMID- 8428370 TI - National Cancer Institute workshop on the possible roles of metallothionein in carcinogenesis. PMID- 8428371 TI - New insight on the biology of neuroectodermal tumors. Workshop report from the University of Rome Tor Vergata and the IDI-IRCCS on the genetics and control of growth, differentiation, and programmed cell death. PMID- 8428373 TI - Signaling at the synapse. PMID- 8428372 TI - Normal and neoplastic growth and development. AACR special conference in cancer research. PMID- 8428374 TI - Neuronal differentiation factors/cytokines and synaptic plasticity. PMID- 8428375 TI - Synaptic circuitry of the retina and olfactory bulb. PMID- 8428376 TI - Developmental mechanisms that generate precise patterns of neuronal connectivity. PMID- 8428377 TI - Synaptic structure and development: the neuromuscular junction. PMID- 8428378 TI - In-vitro recombination in rad and rnc mutants of Saccharomyces cerevisiae. AB - Extracts of S. cerevisiae cells can catalyze homologous recombination between plasmids in vitro. Extracts prepared from rad50, rad52 or rad54 disruption mutants all have reduced recombinational activity compared to wild-type. The rad52 and rad54 extracts are more impaired in the recombination of plasmids containing double-strand breaks than of intact plasmids, whereas rad50 extracts are deficient equally for both types of substrate. The nuclease RhoNuc (previously designated yNucR), encoded by the RNC1 (previously designated NUC2) gene and regulated by the RAD52 gene, is not required for recombination when one substrate is single-stranded but is essential for the majority of recombination events when both substrates are double-stranded. Furthermore, elimination of this nuclease restores recombination in rad52 extracts to levels comparable to those in wild-type extracts. PMID- 8428379 TI - Yeast single copy gene URP1 is a homolog of rat ribosomal protein gene L21. AB - This communication reports on a single-copy gene of Saccharomyces cerevisiae which is homologous to the rat ribosomal protein gene L21. The yeast and the rat genes show 59% identity in DNA sequences and in the predicted protein sequences. This yeast gene is, therefore, assumed to code for an as yet unassigned ribosomal protein (URP1). The URP1 open reading frame is 480 nucleotides long and can encode a protein of about M(r) 18,200. Like most of the other known ribosomal protein genes, URP1 is interrupted by an intron in its 5' terminal part and it is preceded by upstream sequence elements which usually regulate transcription of these genes. Northern blot analysis reveals that the URP1 gene is actually expressed in vivo. PMID- 8428380 TI - Evolutionary conservation of genomic sequences related to the GGP1 gene encoding a yeast GPI-anchored glycoprotein. AB - The GGP1 gene encodes the only GPI-anchored glycoprotein (gp115) that has been purified to-date in the budding yeast Saccharomyces cerevisiae. It is a single copy gene whose deduced amino-acid sequence shares no significant homology to any other known protein. In this paper we report a Southern hybridization analysis of genomic DNA from different eukaryotic organisms to identify homologues of the GGP1 gene. We have analyzed DNA prepared from a unicellular green alga (Chlamydomonas eugametos), from two distantly related yeast species (Candida cylindracea and Schizosaccharomyces pombe), and from the common bean Phasoleus vulgaris. The moderate stringency of the experimental conditions and the high specificity of the probes used indicate that a single-copy of GGP1-related sequences exists in all these eukaryotic organisms. The chromosomal localization of the GGP1 gene in S. cerevisiae has also been determined. PMID- 8428381 TI - Cloning, characterization, and use in strain improvement of the Cephalosporium acremonium gene cefG encoding acetyl transferase. AB - A long open reading frame (ORF) closely linked to the Cephalosporium acremonium gene cefEF was identified by DNA sequencing. The cefEF gene encodes the enzyme involved in cephalosporin C (CPC) biosynthesis known as expandase/hydroxylase. Complementation of a C. acremonium cefG mutant, as well as expression of the gene in Aspergillus niger, showed this ORF to be the cefG gene, encoding cephalosporin C acetyltransferase, which catalyzes the last step in CPC biosynthesis. Analysis of transformants containing additional copies of this gene showed that a direct relationship exists between cefG copy number, cefG message levels, and CPC titers. This gene encodes an enzyme for what may be a rate-limiting step in CPC production. PMID- 8428382 TI - Transformation of four pathogenic Phytophthora spp by microprojectile bombardment on intact mycelia. AB - Phytophthora capsici, P. citricola, P. cinnamomi and P. citrophthora were transformed without the removal of cell walls by particle acceleration with plasmids containing the beta-glucuronidase gene and hygromycin B resistance. Transformants were detected by histochemical and fluorometric beta-glucuronidase assays and confirmed by Southern-blot hybridization. It was found that the promoter of a plant virus is functional in Phytophthora. In addition, a method was designed to visually identify homogeneous transformed colonies, derived from zoospores of transformed multinucleated Phytophthora mycelia, based on blue color development on plates containing X-Gluc. PMID- 8428383 TI - Genetic maps of eight linkage groups of Aspergillus niger based on mitotic mapping. AB - This paper provides a genetic map of Aspergillus niger. At present 84 markers have been assigned to eight linkage groups. The chromosomal location of 60 markers is presented in this paper. The allocation of markers is based on recombination due to mitotic crossing over. Various methods for selection and analysis of homozygous recombinants were applied, using colour, auxotrophic and resistance markers. In addition, transformants carrying the heterologous Aspergillus nidulans gene coding for acetamidase (amdS) were used for mitotic mapping of markers in several linkage groups. In most of the transformants the amdS insert appeared to be centromere-distal to all known genetic markers, thus extending the genetic map. The linear order of the markers in the eight linkage groups has been determined. On the basis of these and earlier experiments tentative genetic maps for the eight linkage groups are presented. Genetic markers were found on both arms of the chromosomes, except for chromosomes II and IV. The genetic distance between markers and the centromere varies from about 10( 4) (LG I, II, V) up to more than 10(-2) (LG III, VI, VIII). The total frequency of mitotic recombination per genome in this fungus has been estimated to be at least 1.2 x 10(-1). PMID- 8428384 TI - A comprehensive compilation of 1001 nucleotide sequences coding for proteins from the yeast Saccharomyces cerevisiae (= ListA2) AB - The amount of nucleotide sequence data is increasing exponentially. We therefore continued our effort to make a comprehensive database for the yeast Saccharomyces cerevisiae. In this database (ListA2) we have compiled 1001 protein coding sequences from this organism. Each sequence has been attributed a single genetic name and in the case of allelic duplicated sequences, synonyms are given, if necessary. For the nomenclature we have introduced a standard principle for naming gene sequences based on priority rules. We have also applied a simple method to distinguish duplicated sequences of one and the same gene from non allelic sequences of duplicated genes. By using these principles we have sorted out a lot of confusion in the literature and databanks. Along with the genetic name, the mnemonic from the EMBL databank, the codon bias, reference of the publication of the sequence and the EMBL accession numbers are included for each entry. The database is available on request. PMID- 8428385 TI - In search of TCR restriction in autoreactive T cell in human autoimmunity: why is it so elusive? PMID- 8428386 TI - Common idiotypes expressed on human, monoclonal, abnormal immunoglobulins and cryoglobulins with polyreactive autoantibody activities. AB - Several human monoclonal immunoglobulins with the same autoantibody activity have been shown to have cross-reactive idiotypes (CRI). In this study, using polyclonal anti-idiotypic antibodies, we found that 28% of human monoclonal immunoglobulins with polyreactive autoantibody activity from myeloma, Waldenstrom's macroglobulinaemia and cryoglobulinaemia patients shared common idiotype(s). Furthermore, the latter were expressed on human and murine natural MoAbs (respectively in 12% and 22% of the clones tested) and on human IgG preparations used for therapeutic intravenous injections (IVIg) and which contain natural antibodies. These findings suggest that monoclonal immunoglobulins could arise from the proliferation of a clone that normally produces a natural antibody. The existence of common idiotype(s) between monoclonal immunoglobulins and IVIg could be relevant to the improvement noted after treatment with IVIg in patients suffering from peripheral neuropathies associated with monoclonal gammopathy. PMID- 8428387 TI - A high proportion of the V delta 1+ synovial fluid gamma delta T cells in juvenile rheumatoid arthritis patients express the very early activation marker CD69, but carry the high molecular weight isoform of the leucocyte common antigen (CD45RA). AB - We have previously shown that gamma delta T cells in the synovial compartment of patients with juvenile rheumatoid arthritis (JRA) express activation antigens (CD69 and HLA-DR) and that they are predominantly of the V delta 1 subset. In this study we have analysed the expression of activation antigens (CD69 and HLA DR) and different isoforms of the leucocyte common antigen (CD45RO and CD45RA) on the V delta 1 and the V delta 2 subsets of gamma delta T cells in paired samples of synovial fluid and peripheral blood of nine patients with JRA, and in the peripheral blood of five children with idiopathic scoliosis. In the synovial fluid of children with JRA, there were significantly more V delta 1+CD69+ and V delta 2+CD69+ cells compared with the peripheral blood of the same patients. In contrast, however, in the synovial fluid the V delta 1 and the V delta 2 subsets differed with respect to the expression of the two isoforms of the leucocyte common antigen. The majority of the V delta 1+ cells expressed the high molecular weight isoform (CD45RA+) while most of the V delta 2+ cells carried the low molecular weight variant (CD45RO+) of this molecule. PMID- 8428388 TI - HLA-B27 subtypes in the spondarthropathies. AB - The spondarthropathy (Sp)-associated HLA-B27 antigen includes at least seven subtypes, B*2701-07, of which 01, 02, 05 and 07 occur in Caucasians. This study examined the B27 subtype distribution in British patients with Sp. The 133 HLA B27+ subjects comprised 94 European Caucasian Sp (58 ankylosing spondylitis (AS), 22 reactive arthritis (ReA; 11 sexually acquired (SARA), 11 enteric (EReA)), eight undifferentiated Sp (USp), and six pauciarticular juvenile-onset chronic arthritis (pJCA)) patients, and 34 healthy Caucasian controls, together with four Asian Indian and one Chinese. 35S-labelled B27 was immunoprecipitated with anti B27 MoAbs, and subtyped according to isoelectric point (pI) following isoelectric focussing. The use of B27 MoAb permitted subtype assignment without full class I HLA typing. The vast majority (95%) were B*2705 (Caucasian controls 31/34; AS 55/58; ReA 21/22; USp 8/8, and pJCA 6/6; Indian control 1/1 and AS 2/3; Chinese pJCA 1/1), and the remainder B*2702. No B*2701 or 07 subjects were identified. AS occurs in both B*2702 and 05 subjects, and we extend this observation to small numbers of ReA and of Indian AS subjects. This implicates molecular features shared between B27 subtypes, rather than subtype-determining regions of the antigen, in Sp pathogenesis. PMID- 8428389 TI - Macrophage depletion decreases IgG anti-DNA in cultures from (NZB x NZW)F1 spleen cells by eliminating the main source of IL-6. AB - We have studied the role of macrophages in the production of IgG anti-DNA autoantibodies by (NZB x NZW)F1 mice (B/W). One of the main features of the systemic lupus erythematosus (SLE)-like disease that affects these mice, is the presence of circulating IgG autoantibodies and immune complexes, which lead to renal failure and death by the age of 8-9 months. IgG autoantibodies are produced without in vitro stimulation by total spleen cells from these mice when they reach the age of 6 months. We have demonstrated that IL-6 increases the production of IgG autoantibodies in cultures of splenic purified B cells from the old B/W mice. The aim of this study was to show the involvement of macrophages in the production of IL-6 and consequently in the production of IgG anti-DNA antibodies in vitro. We show that elimination of the macrophages by different treatments led to reduction of the content of IL-6 in the supernatants as well as of IgG anti-DNA autoantibodies. Addition of fresh, splenic or peritoneal macrophages restored the production of autoantibodies in macrophage-depleted cultures from old B/W mice. There were no differences in the capacity of IL-6 production between macrophages from old or young B/W mice, but an important difference was observed between peritoneal and splenic macrophages, where the former produced much higher levels of IL-6, and consequently were more potent inducers of IgG autoantibodies. The present results reinforce the role of macrophages and IL-6 in the production of IgG anti-DNA autoantibodies in B/W mice. The implications of these results in the pathogenesis of the disease are discussed. PMID- 8428390 TI - Prevention of cyclophosphamide-induced diabetes in the NOD/WEHI mouse with deoxyspergualin. AB - Ten out of 20 (50%) 17-week-old female NOD/WEHI mice developed an acute form of autoimmune diabetes when injected with two large doses of cyclophosphamide (CY), given at 14-day intervals. If these mice were treated under a prophylactic regimen with 2.5 mg/kg body weight per day of the novel immunosuppressant deoxyspergualin (DSP) the onset of diabetes was completely prevented. Moreover, DSP-treated animals showed reduced signs of pancreatic insulitis, had lower percentages of splenic lymphoid cells (SLC) expressing IL-2 receptors and Ly-6C antigens on their surfaces, and these cells released lower amounts of interferon gamma (IFN) when stimulated in vitro. These data, providing evidence for the capacity of DSP to protect NOD/WEHI mice from experimental autoimmune diabetes and to modulate histo-immunological pathogenic pathways, indicate DSP as a drug of potential interest in the treatment of human insulin-dependent diabetes mellitus. PMID- 8428391 TI - Defective IL-6 secretion in HIV-infected haemophilia patients. AB - To study the role of IL-6 in HIV-induced B cell defects, in vitro B cell responses and IL-6 secretion were determined simultaneously in 67 haemophilia patients. Twenty-three patients were HIV- (Group 1), 27 HIV+ stage CDC II, III (Group 2), and 17 were HIV+ stage CDC IV (Group 3). Pokeweed mitogen (PWM) was used for T cell-dependent and Staphylococcus aureus Cowan I (SAC I) for T cell independent B cell stimulation. B cell differentiation was assessed in a reverse haemolytic plaque assay and by ELISA determination of IgG and IgM in culture supernatants. An ELISA was used to measure IL-6 in plasma and culture supernatants. HIV- patients showed impaired immunoglobulin-secreting cell (ISC) responses after T cell-independent and T cell-dependent stimulation (P < 0.0001 and P < 0.01, respectively), whereas IL-6 secretion, IgM and IgG responses were comparable to those in healthy controls. HIV+ patients at stage CDC II, III or IV demonstrated significantly reduced mitogen-stimulated IL-6 secretion (P < 0.05, PWM; P < or = 0.001, SAC I) as well as impaired ISC and IgG responses (P < 0.01, PWM; P < or = 0.0001, SAC I). CDC IV patients showed reduced IgM responses in addition (P < 0.02, PWM; P < 0.0005, SAC I). Plasma IL-6 levels were elevated both in HIV+ patients (CDC II, III patients: 165 +/- 73 pg/ml, P < 0.005; CDC IV patients: 58 +/- 18 pg/ml, P < 0.0001) and in HIV- patients (283 +/- 65 pg/ml, P < 0.0001) which appeared to be a T cell effect induced by treatment with haemophilia factor concentrates. Our data provide evidence for different types of B cell deficiencies in HIV- patients (impaired ISC response only) and HIV+ patients (impaired ISC as well as IL-6 and IgM/IgG responses). The defective IL-6 secretion in HIV+ patients is likely to affect terminal B cell differentiation and this may explain the reduced immunoglobulin secretion in these patients in response to antigenic challenge. PMID- 8428392 TI - Differential mechanisms of intracellular killing of Mycobacterium avium and Listeria monocytogenes by activated human and murine macrophages. The role of nitric oxide. AB - Murine peritoneal macrophages activated with interferon-gamma (IFN-gamma) produce large quantities of nitric oxide and are efficient in the killing of certain intracellular pathogens. To examine the role of this mechanism in the killing of Mycobacterium avium by murine and human macrophages, we infected mouse peritoneal macrophages and human monocyte-derived macrophages with M. avium and Listeria monocytogenes and stimulated the cells with recombinant tumour necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF) or IFN-gamma, in the presence or absence of N-monomethyl-L-arginine (NMA) or arginase. Neither competitive inhibition with NMA nor depletion of arginine by arginase had any effect on the inhibition of growth/intracellular killing of M. avium by activated human and murine macrophages. In contrast, activation of murine but not human macrophages infected with L. monocytogenes by IFN-gamma was significantly inhibited by the addition of NMA/arginase. Furthermore, murine macrophages produced large concentrations of nitric oxide following stimulation with recombinant cytokines, although no significant increase of nitric oxide production was observed with human monocyte-derived macrophages. PMID- 8428393 TI - Differential cytokine gene expression and secretion after phagocytosis by a human monocytic cell line of Toxoplasma gondii compared with Mycobacterium tuberculosis. AB - Toxoplasma gondii infection may be clinically silent in immunocompetent individuals but may cause fatal disease in immunocompromised patients such as those with HIV infection. Proinflammatory cytokines are known to be important in murine resistance to T. gondii but there are no data from human models of infection. We have investigated whether phagocytosis of T. gondii, of Mycobacterium tuberculosis (a pathogen which elicits a granulomatous host immune response) and of inert latex particles by THP-1 cells, a human monocytic line, caused gene expression and secretion of tumour necrosis factor (TNF), IL-6 and IL 8. These cytokines are important in recruitment and activation of T lymphocytes, and both TNF and IL-6 may have direct antitoxoplasmacidal and antimycobacterial activity. Phagocytosis of T. gondii by THP-1 cells resulted in minimal gene expression and secretion of TNF, IL-6 and IL-8 similar to that following phagocytosis of inert latex particles. In contrast, phagocytosis of M. tuberculosis resulted in increased gene expression of TNF and IL-8 as well as increased secretion of all three cytokines, particularly IL-8. These observations may partially explain the frequency of non-inflammatory host responses to T. gondii in immunocompetent individuals. PMID- 8428394 TI - Cytokine-induced inhibition of Plasmodium falciparum erythrocytic growth in vitro. AB - The addition of recombinant cytokines to Plasmodium falciparum in vitro cultures retarded the growth of the parasite with the effect of recombinant IL-2 (rIL-2) > interferon-gamma (IFN-gamma) > tumour necrosis factor-beta (TNF-beta). The process was concentration dependent, being greatest at 30,000 U/ml and required a 72-h period of continuous exposure for maximum effect. Growth inhibition, as determined morphologically and radiometrically, was a consequence of defective schizont maturation rather than inhibition of merozoite invasion. It was cumulative and detectable within one erythrocytic (48 h) growth cycle. PMID- 8428395 TI - Exposure to complement-bearing immune complexes enhances the in vitro sequestration of erythrocytes from young but not elderly donors. AB - Complement and immunoglobulin have each been claimed to be the major opsonins responsible for sequestration of the effete erythrocyte. Binding of immune complexes to the erythrocyte via CR1 (CD35) provides a model for studying the effects of increments in membrane-bound complement and immunoglobulin on the sequestration of the erythrocyte ('innocent bystander' sequestration). It is possible that C3b-bearing immune complexes (IC-C3b) bound to erythrocyte CR1 contribute to the levels of immunoglobulin and complement fragments detectable on the human erythrocyte. We have, therefore, compared the capacity of erythrocytes from young and elderly donors to bind IC-C3b and the effect of this binding on in vitro sequestration. Erythrocytes from young donors exposed to IC-C3b bind these complexes, as attested by an increment in membrane-bound C3, and undergo 'innocent bystander' sequestration. However, when density-separated erythrocytes are so exposed, it is only the low density (young) erythrocytes from young donors which are susceptible to 'innocent bystander' sequestration. High density (old) erythrocytes from young donors and all erythrocytes from elderly donors show initially high in vitro sequestration and are resistant to the 'innocent bystander' effect. (Those erythrocytes which show initially high in vitro sequestration are referred to collectively as 'in situ aged' erythrocytes.) There is a great similarity between the mechanisms of sequestration of 'in situ aged' and 'innocent bystander' erythrocytes in that they are both inhibited by the integrin binding peptide arginine-glycine-aspartic acid (RGD) and the beta galactosyl sugar N-acetyl-galactosamine, and unaffected by the Fc-gamma binding protein, Protein-G. Complement is the major opsonin in 'innocent bystander' sequestration since this sequestration occurs whether the isotype of the antibody in the immune complex is IgM or IgG, and Protein-G, which inhibits IgG-dependent erythrophagocytosis, has no effect on 'innocent bystander' sequestration. The present studies demonstrate that in vitro sequestration of 'in situ aged' erythrocytes is similar to complement-dependent 'innocent bystander' sequestration, thus supporting the contention that complement recognition is cardinal in sequestration of 'in situ aged' erythrocytes. PMID- 8428396 TI - Combinations of low concentrations of cytokines and acute agonists synergize in increasing the permeability of endothelial monolayers. AB - The deposition of circulating immune reactants in blood vessels, an important event in the pathogenesis of certain types of vasculitis, requires an increase in permeability in the endothelial monolayer. An in vitro model to examine the integrity of endothelial cell monolayers and their response to inflammatory mediators has been developed. Human umbilical vein endothelial cells were grown to confluence on an FITC-labelled matrix and monolayer integrity was assessed by the exclusion of a 125I-anti-FITC antibody. Alteration in endothelial monolayer permeability was associated with an increase in uptake of 125I-anti-FITC antibody, expressed as a percentage of the maximal uptake of antibody on to FITC matrix from which endothelial cells had been stripped. We determined the effects on endothelial monolayer permeability of acute agonists (thrombin and histamine), cytokines (tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN gamma), IL-1 and IL-4) and combinations of acute agonists and cytokines. Addition of thrombin in concentrations ranging from 0.5 to 15 U/ml led to an increased uptake of 125I-anti-FITC antibody from 2% to 15% relative to unstimulated endothelium. For other agonists and cytokines the increases in permeability were: (i) histamine (50-400 pmol/ml) increased uptake 5-22%; (ii) TNF (12.5-100 ng/ml) increased uptake 2-12%; (iii) IFN-gamma (125-250 U/ml) increased uptake 1.5-3%. IL-1 beta (50-100 U/ml) and IL-4 (50-100 U/ml) had no effect. Synergistic interactions on endothelial monolayer permeability were seen with the following combinations: (i) IL-4 (100 U/ml) and TNF (12.5 ng/ml) uptake 11%; (ii) IL-4 (100 U/ml) and IFN-gamma (125 U/ml) uptake 6.5%; (iii) TNF (12.5 ng/ml) and IFN-gamma (125 ng/ml) uptake 7%; (iv) thrombin (0.5 U/ml) and histamine (50 pmol/ml) uptake 13.5%; and (v) TNF (12.5 ng/ml) and thrombin (0.5 U/ml) uptake 8.5%. These observations suggest that interactions between cytokines and acute inflammatory mediators such as thrombin and histamine may be important in determining whether immune complexes are deposited in vessel walls. This model system may now be useful for the further investigation in vitro of the mechanisms involved in the pathogenesis of immune complex-mediated vascular damage. PMID- 8428397 TI - IL-1 beta and IL-3-like activity in preterm infants. AB - The capacity of peripheral blood mononuclear cells (PBMC) of preterm neonates to release IL-1 beta and IL-3-like activity (IL-3-LA) has been investigated. In the present study it was found that this capacity is significantly lower than that of their mothers and of control adults. In addition, the results showed that preterm serum has a lower stimulatory effect on IL-1 beta production and an inhibitory effect on IL-3-LA secretion by PBMC of adult controls, in comparison with maternal and adult sera. These findings suggest an additional feedback mechanism for control of haematopoiesis in premature neonates. It is possible that the lower production of IL-1 beta and IL-3-LA may be involved in the increased susceptibility to infections of preterm newborns. PMID- 8428398 TI - Induction of mouse monocyte Ia expression by a mesangial cell-derived product. AB - Previous studies in many laboratories have shown that macrophage Ia expression is not constitutive but under regulation. We provide data which demonstrate that product(s) of mouse mesangial cell cultures induce blood monocyte Ia expression as demonstrated by immunofluorescence. This process is time-related and is also dependent on novel protein synthesis, being abrogated when the monocytes are pretreated with cycloheximide. Preliminary characterization shows the mesangial cell product to be sensitive to heating at 100 degrees C x 30 min, to be resistant to digestion by trypsin at a concentration of 4 x 10(-6) M, and to have a molecular size of 10-100 kD as established by Amicon ultrafiltration. The substance is not interferon-gamma (IFN-gamma) since cultured mesangial cells had no contaminating T cells, mesangial cell supernatant had no detectable levels of IFN-gamma, and the Ia-inducing activity of the mesangial cell product was not abrogated by incubation of monocytes with mesangial cell supernatant which had been immunoprecipitated with anti-IFN-gamma. Similarly, experiments using anti CSF-1 have excluded the possibility that the substance is CSF-1. The results of the study have relevance to the mechanisms by which monocytes which take up residence in the glomerular mesangium acquire Ia positivity, and also provide a potentially novel pathway by which a tissue product may induce monocytes to express Ia. PMID- 8428399 TI - Direct evidence that human follicular dendritic cells (FDC) rescue germinal centre B cells from death by apoptosis. AB - It is supposed that FDC have a pivotal role in the rescue of germinal centre (GC) B lymphocytes from apoptosis. However, formal proof for this hypothesis has not as yet been presented. In the present study FDC and GC B cells were isolated from human tonsils and cultured. When brought into culture FDC and B cells rapidly formed spherical clusters. T cells were not observed inside these clusters. At different time points cultures of FDC and B cells were supravitally stained with Hoechst 33258 or acridine orange and examined by direct observation using fluorescence microscopy. Viable B cells appeared to be profoundly restricted to clusters, whereas cells not taking part in clusters all had an apoptotic appearance. The formation of clusters could be prevented by addition of MoAbs against CD11a (LFA-1 alpha) or CD49d (VLA-4 alpha), resulting in an apoptotic appearance of virtually all B lymphocytes. The present data demonstrate that a physical interaction between FDC and germinal centre B lymphocytes is able to rescue the latter from apoptotic cell death. PMID- 8428400 TI - Percentile ranges for serum IgG subclass concentrations in healthy Chinese children. AB - In order to establish normal reference ranges of serum immunoglobulin G subclasses for different age groups in Chinese, we measured the four IgG subclasses by radial immunodiffusion using polyclonal antisera in 350 normal healthy subjects (148 males and 202 females) recruited from the community. There was no significant sex difference for all the four IgG subclasses. Using Box-Cox transformation, we constructed smooth age-dependent percentile curves for the four IgG subclasses. For the 50th percentile values, the plateaus for IgG1 to IgG4 are respectively 970 mg/dl at 13 years old, 481 mg/dl at 18, 48 mg/dl at 17 and 80 mg/dl at 13. Methodology, reagents, environmental and genetic factors might be responsible for the observed difference among the various reports. The IgG2 level in our Chinese population seemed to be higher than that in Caucasians, which might account for the very low incidence of invasive Haemophilus influenzae type b disease among the Chinese. Our IgG1 level also plateaued later and at a higher level than that in other studies. PMID- 8428401 TI - Ibuprofen as an over-the-counter drug: is there a risk for renal injury? AB - Ibuprofen has recently been introduced as over-the-counter drug in several countries. The question has been raised whether or not the increased availability and use of ibuprofen as over-the-counter drug will be associated with an increased incidence of nephrotoxic side-effects. It is unquestionable that ibuprofen can cause renal damage, including functional acute renal failure, water and electrolyte disorders, and interstitial nephritis. Analgesic nephropathy is not a documented consequence of ibuprofen therapy. Renal side-effects of ibuprofen appear to be dose-dependent, and were not reported at the recommended dosage as over-the-counter drug (0.2-0.8 g/d) except for a single child. Even at anti-inflammatory doses (> 1.6 g/d), renal side-effects are almost exclusively encountered in patients with low intravascular volume and low cardiac output particularly in old age. Alternative analgesic and antipyretic agents display no less risk for renal injury than ibuprofen. PMID- 8428402 TI - Abnormal urinary protein excretion in HIV-infected patients. AB - HIV infection has been associated with a variety of renal diseases, although the pathogenesis of such dysfunction is unknown. To determine whether HIV-infection is associated with glomerular permeability defects, and if so, the prevalence of the finding, we studied patients with various stages of HIV infection. Urine samples from 505 outpatients with HIV infection (without hypertension, azotemia, or dipstick proteinuria), 41 normal controls and 40 febrile non-HIV positive, hospitalized patients with infectious diseases were analyzed for the urinary microalbumin/creatinine ratio (U microA/Cr), a sensitive indicator of incipient renal disease in diabetes mellitus and hypertension, and the urinary beta 2 microglobulin/creatinine ratio (U beta 2/Cr), an indicator of renal tubular function. Microalbumin concentration was measured by ELISA. Beta 2-microglobulin concentration was measured by an enzyme immunoassay. HIV-infected outpatients had higher mean U microA/Cr than normal subjects, but not febrile hospitalized controls. The prevalence of an increased U microA/Cr was 29.8% in the HIV infected outpatient population. There was no difference in the ratio between Black and White HIV-infected outpatients, HIV-infected outpatients treated or untreated with zidovudine (AZT), or HIV infected outpatients untreated with any drug. There was no difference between U microA/Cr in stage II, III or IV HIV infected patients when assessed by analysis of variance. A similar pattern was noted with U beta 2/Cr. The prevalence of an increased U beta 2/Cr ratio was 37.7% in HIV-infected outpatients. Increased urinary albumin and beta 2 microglobulin excretion, not associated with drug therapy, is present in patients with early HIV infection.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428403 TI - Calcium metabolism in acute renal failure due to rhabdomyolysis. AB - We report a patient with drug and hyperthermia induced rhabdomyolysis who developed acute renal failure. During the oliguric phase of 22 days, there was profound hypocalcemia (lowest ionized calcium of 0.34 mmol/l), associated with appropriately elevated intact PTH levels and high normal 1,25(OH)2D levels. Massive calcification in necrotic muscle occurred during this time. In the recovery phase, hypercalcemia was present lasting 33 days (maximum ionized calcium of 1.99 mmol/L), associated with suppression of PTH secretion, low 1,25(OH)2D3 levels, decreased bone resorption and mobilization of the muscle calcium deposits. This case report illustrates that the changes in serum calcium in rhabdomyolysis-associated acute renal failure are explicable by the deposition or removal of mineral into or from necrotic muscle with the parathyroid and vitamin D changes occurring secondarily. PMID- 8428404 TI - Chronic renal failure due to renal hemosiderosis in a patient with paroxysmal nocturnal hemoglobinuria. AB - We report on a 44-year-old man with paroxysmal nocturnal hemoglobinuria, who developed renal hemosiderosis, interstitial nephritis, and chronic renal failure. Classically, it is assumed that the renal function is only uncommonly affected by this pathology. In most instances the renal hemosiderosis is an incidental finding at postmortem examination. In our case the presence of iron in the kidney, as confirmed by CT scan, magnetic resonance imaging and renal biopsy, occurred concomitantly with the development of renal insufficiency. A direct nephrotoxic effect of iron by the induction of highly reactive hydroxyl radicals is suggested. PMID- 8428405 TI - Use of transdermal clonidine in chronic hemodialysis patients. AB - Large fluctuations of blood pressure are commonly experienced by hypertensive, chronic hemodialysis patients. Many patients hold anti-hypertensive medication immediately prior to dialysis to prevent intradialytic hypotension. Weekly dosing of continuously released antihypertensive agents may result in better blood pressure control than conventional daily dosing. To evaluate the effect of weekly transdermal clonidine on this problem, we compared intra- and interdialytic blood pressure control and side effects during six weeks of transdermal clonidine monotherapy and six weeks of conventional oral antihypertensive treatment in nine stable chronic hemodialysis patients. Since transdermal clonidine is recommended for mild to moderately severe hypertension and since clonidine is excreted by the kidneys and removed by hemodialysis, we also evaluated blood pressure control and clonidine levels while utilizing high-dose transdermal clonidine, up to 0.12 mg per week. Intradialytic blood pressure was monitored twice weekly during the weeks 3-6 of transdermal clonidine and conventional therapy. Twenty-four hour blood pressure was monitored weeks 3 and 6 of each study phase. Plasma clonidine levels were measured by HPLC in 11 patients. Transdermal clonidine monotherapy failed to adequately control blood pressure in 6 of 21 chronic dialysis patients with moderate to severe hypertension. No significant difference in intra- or interdialytic blood pressure control or side effects (including dry mouth or thirst) was found comparing conventional to transdermal clonidine therapy. While not statistically different, heart rate was lower during transdermal clonidine therapy compared to conventional therapy, especially at very high doses. Despite a mean hemodialysis clearance of clonidine of 59.2 +/- 7.8 ml/min, clonidine levels remained therapeutic beyond one week.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428406 TI - The use of transdermal clonidine in the hypertensive patient with chronic renal failure. AB - The efficacy of transdermally administered clonidine was evaluated in twenty-four patients with mild-to-moderate hypertension (seated diastolic blood pressure 95 115 mmHg) and renal impairment. Patients were initially treated with oral clonidine; the dose was titrated until the seated diastolic pressure fell below 90 mmHg or a minimum 10% reduction in baseline was achieved. Oral clonidine produced a significant decrease in both systolic and diastolic blood pressure; mean seated diastolic blood pressure decreased 16.9 mmHg from baseline. When transdermal clonidine was substituted for twice daily dosage of oral clonidine, blood pressure decreases were fully maintained. Sixteen patients completed three months of stable-dose transdermal therapy. The results suggest that, in mild-to moderate hypertensive patients with chronic renal impairment, blood pressure can be controlled with a once-weekly application of transdermal clonidine as effectively as with oral clonidine. Mean diastolic blood pressure was decreased approximately 17% during clonidine therapy independent of the severity of renal dysfunction or route of clonidine administration. PMID- 8428407 TI - The use of urinary red cell morphology to determine the source of hematuria in children. AB - Urinary red cell morphology has been used to indicate the source of renal tract bleeding. A double blind study was undertaken to evaluate the reliability of urinary red cell morphology in predicting the source of hematuria in a pediatric population. Two independent observers used phase-contrast microscopy, one also using Nomarski differential interference microscopy, to examine 101 urine specimens from 90 pediatric patients with hematuria. 28 cases were subsequently excluded because the clinical diagnosis was uncertain. In the remaining 62 patients the laboratory diagnosis based on the erythrocyte morphology was compared with the primary clinical diagnosis. Observer 1 documented dysmorphic red cells in 40 of 42 patients with glomerulonephritis, and isomorphic red cells in 19 of the 20 cases of non-glomerular bleeding. Observer 2 documented dysmorphic red cells in 39 of the 42 glomerular lesions, and isomorphic red cells in all of the non-glomerular lesions. This represents a sensitivity for predicting glomerular hematuria of 95% for observer 1 and 93% for observer 2, and a specificity of 95% and 100% respectively. There was no definite advantage in the use of Nomarski compared to phase-contrast microscopy. These results confirm the accuracy of this technique in predicting the source of hematuria in pediatric patients. PMID- 8428408 TI - A new method of evaluating urinary erythrocyte dysmorphisms (UED) in glomerulonephritis. AB - A new method of evaluating urinary sediment, based on electronic processing of the images obtained by traditional microscopy, has been developed. Preliminary data suggests that the method can clarify many doubts on the real diagnostic value of UED. PMID- 8428409 TI - Pharmacokinetics of ciprofloxacin in subjects with varying degrees of renal function and undergoing hemodialysis or CAPD. AB - The pharmacokinetics of a single, oral dose of 750 mg of ciprofloxacin were studied in 35 subjects with various degrees of renal function (Group 1, Clcr > or = 80 ml/min; Group II, Clcr 50-79 ml/min; Group III, Clcr 10-49 ml/min) and on hemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD). Blood, urine and CAPD dialysate samples were collected over a period of 48 hours after dosing. Data were fitted using non-linear, least squares regression. The mean Cmax was 3.4 +/- 1.0 mg/l and tmax was 2.3 +/- 0.9 hours. The mean AUC in Group I was 14.7 mg.h/l, Group II was 33.7 (p < 0.001), Group III 63.8 (p < 0.001), HD 57.9 (p < 0.0001) and CAPD 44.3 (p < 0.001). Half-life in Group I was 4.6 h, and was shorter than Group III (11.1 h, p < 0.001), HD (13.4 h, p < 0.001) and CAPD (8.9 h, p < 0.001). Total body clearance and renal clearance demonstrated significant differences also. The dialysis clearance in CAPD patients was 0.53 +/ 0.39 l/h. Peritoneal effluent concentrations varied from 0.6 mg/l during the first exchange, to a peak of 2.2 mg/l during the second, to 0.13 mg/l in the 48 hour (9th) exchange. Dosage adjustments of ciprofloxacin in the presence of renal insufficiency are indicated for subjects with a Clcr < 20 ml/min/1.73m2. PMID- 8428410 TI - Nature of progressive glomerulosclerosis in human membranous nephropathy. AB - In membranous nephropathy (MN) the mechanism of progression is not well defined. We studied the lesions of focal segmental glomerulosclerosis (FSGS) in 95 patients with idiopathic MN in relation to other morphologic and clinical data. Forty-one patients (43%) showed FSGS, frequently accompanied by synechiae and hyalinosis. The patients with FSGS had a significantly greater degree of mesangial expansion, glomerular basement membrane (GBM) thickening, interstitial fibrosis, and arteriolosclerosis, as well as an increased level of serum creatinine, when compared to the patients without. Relative interstitial volume, in particular, significantly correlated to the serum creatinine level, thickness of GBM and % of glomeruli with FSGS. Glomerular size was not different in patients with or without FSGS. When clinical and morphological data of these patients were analyzed according to a staging technique of Ehrenreich and Churg [1968], the occurrence of FSGS was more frequently present in advanced stage. Intraglomerular fat deposition was only noted in advanced stage. No other morphologic or clinical parameters were related to the staging. Thickness of GBM or index of electron density of deposits, though well related to the morphological staging, did not show any relationship to clinical or laboratory data. Sixty-four patients were followed for more than a year (mean 3 years). Patients with FSGS had more frequent progression compared with those without. Patients with advanced stage and FSGS did not do worse than patients with stage I II+II and FSGS. From these observations, we believe that the mesangial expansion, advanced GBM thickening, interstitial fibrosis and arteriolosclerosis are important morphological parameters, which may play a role in the genesis of FSGS or progression in MN. In addition, the occurrence of FSGS, but not the morphological staging, can predict the course of individual patients in MN. PMID- 8428411 TI - CT portography and delayed high dose iodine CT. PMID- 8428412 TI - Wire localized biopsy of breast lesions: a review of 425 cases found in screening or clinical mammography. AB - A total of 425 wire localized biopsies for non-palpable breast lesions found by screening (n = 169) or clinical (n = 256) mammography between 1986-1990 are reviewed with reference to the biopsy success rate with different localization techniques, the positive predictive value of the mammographic findings and the staging and treatment of carcinomas in the clinical and screening groups. Specimen radiographs were available for review in 371 cases and confirmed the removal of the lesion in 90.6%, were indeterminate in 4.8% and were negative in 4.6%. Some 95% of the failures occurred in mammography-guided localizations using the Kopans hookwire. In later years this wire was replaced with th Homer retractable needle/wire system, with a 99% success rate. The Kopans wire was found to be accurate in ultrasound (US)-guided procedures, which were tolerated very well with no complications, as compared with a 10.2% rate of vasovagal reactions in mammography-guided procedures. The success rate of the localizations performed under general anaesthesia was 93.7%, as compared with 80.2% of the localizations performed under local anaesthesia. A more clearcut improvement of the success rate (from 66.7% in 1986-1987 to 99% in 1990) was seen over the years with growing experience and the development of teamwork between radiologists, surgeons and pathologists. The histological diagnosis was malignant in 27% of cases. The positive predictive value for malignancy improved from 21% to 32% from 1986 to 1990, due mainly to an increase in the screening group. A total of 71% of the carcinomas found were of stage 0 (TisN0M0) or I (TIN0M0) indicating a good prognosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428413 TI - The computed tomographic appearances in chronic berylliosis. AB - Computed tomography (CT) of the thorax was performed in eight patients with chronic berylliosis. The pulmonary CT features were variable and included pulmonary nodularity, and patterns consistent with upper lobe fibrosis or diffuse interstitial fibrosis. Mediastinal or hilar lymphadenopathy was shown in three cases. CT clearly demonstrates the pattern and distribution of pulmonary and mediastinal involvement in chronic pulmonary berylliosis but the appearances are non-specific. PMID- 8428414 TI - The role of percutaneous nephrostomy in malignant urinary tract obstruction. AB - Percutaneous nephrostomy is a well established technique for rapid relief of ureteric obstruction and improvement of renal function. However, its role in the management of renal failure resulting from advanced abdominopelvic malignancy is controversial and there are no clear guidelines to predict which patients benefit from such intervention both in terms of survival time and quality of life. To establish a protocol for selection of patients with abdominopelvic malignancy most likely to benefit from nephrostomy for renal obstruction, the medical records of 50 consecutive patients undergoing this procedure at the Royal Marsden Hospital were reviewed. The patients were divided into four groups: Group I, renal obstruction caused by a nonmalignant complication as a result of previous surgery or radiotherapy (n = 8); Group II, renal obstruction due to untreated primary malignancy (n = 16); Group III, renal obstruction from relapsed disease with a viable treatment option (n = 8); and Group IV, relapsed disease with no conventional treatment option (n = 18). There was significant benefit from percutaneous nephrostomy in Groups I-III. The overall median survival time of Group IV patients was extremely poor: 38 days (range 6-143 days) with no long term survivors. The results suggest that strict selection criteria should be applied to patients with a history of abdominopelvic malignancy before proceeding to percutaneous nephrostomy. No worthwhile benefit is obtained if nephrostomy is used as a palliative measure in the absence of definitive treatment. PMID- 8428415 TI - Ultrasound findings in hepatic mycobacterial infections in patients with acquired immune deficiency syndrome (AIDS). AB - Ultrasound findings in 12 AIDS patients with abdominal mycobacterial infections were reviewed and correlated with liver histology. Liver ultrasound abnormalities were common--present in 4/5 patients with Mycobacterium avium-intracellulare (MAI) and 7/7 patients with Mycobacterium tuberculosis (MTB) infection. The commonest ultrasound abnormality of the liver was a generally 'bright' liver, seen in 7/12 patients. Focal liver lesions were seen in 5/7 patients with MTB but were not seen in any patients with MAI infection. Both hyperechoic (two patients) and hypoechoic (three patients) lesions were seen. Lymphadenopathy as demonstrated on abdominal ultrasound was a relatively infrequent finding--only seen in three patients with MTB, all of whom also had focal liver lesions. On histology, 8/12 patients showed fatty infiltration and 8/12 showed granuloma. Abnormalities are commonly seen on ultrasound examination of the liver in AIDS patients with abdominal mycobacterial infections but are non-specific and ultrasound guided biopsy is indicated to confirm the diagnosis and exclude other disease. PMID- 8428416 TI - Phosphor plate computed radiography: response to variation in mAs at fixed kVp in an animal model. Potential role in neonatal imaging. AB - Computed digital radiography offers a number of potential advantages over conventional film-screen imaging. To determine the applicability of these to portable neonatal examinations we performed a controlled prospective study comparing conventional 200 speed film-screen with a computed radiography unit in an animal model. Images acceptable for diagnostic purposes were obtained on the digital system over an exposure range many orders of magnitude greater than was possible on the film-screen system. Digital imaging offers potential for elimination of repeat examinations due to suboptimal exposure factors, and for reduction in radiation dose to the patient. We believe that computed digital radiography should be particularly suited to portable neonatal imaging. PMID- 8428417 TI - The small bowel enema: a ten year review. AB - In a retrospective study the radiological reports of small bowel enema (enteroclysis) examinations of 1465 patients were reviewed and compared with the subsequent clinical outcome, and where possible with findings at laparotomy. A sensitivity of 93.1% and a specificity of 96.9% was found, based on whether the small intestine was reported as normal or an abnormality was diagnosed to account for the patient's clinical presentation. The correct specific diagnosis was made in 67.5% of the examinations that were considered abnormal. We believe that these figures support the use of small bowel enema as the routine barium examination for suspected disorders of the small intestine. PMID- 8428418 TI - Perineal herniation. AB - In a retrospective study of 800 evacuating proctograms, 37 patients were found to have a varying degree of perineal herniation. The radiological and surgical correlation of this interesting abnormality was discussed. PMID- 8428419 TI - Premature tracheobronchial, laryngeal and costochondral cartilage calcification in children. AB - Two children with premature tracheobronchial, laryngeal and costochondral cartilage calcification are described. Both patients had associated congenital cardiovascular anomalies; one of them having features of Keutel syndrome. PMID- 8428420 TI - Normal symphysis pubis width in children. PMID- 8428421 TI - Case report: thyroid acropachy in a single digit. AB - A case of florid thyroid acropachy affecting a single bone is described. Its potential to mimic malignancy is illustrated. PMID- 8428422 TI - Case report: unusual computed tomographic appearances of meningioma initially misdiagnosed as an intrinsic tumour. PMID- 8428423 TI - Case report: the ultrasound appearances of testicular microlithiasis ('snow storm' testis): a case complicated by testicular seminoma. AB - A case is described in which a patient with bilateral testicular seminomas was found also to have testicular microlithiasis. The ultrasound and pathological features are described. PMID- 8428424 TI - Transaxillary arteriograms. PMID- 8428425 TI - Fetal medicine and ultrasonography. PMID- 8428426 TI - Staging of cervical and endometrial carcinoma. PMID- 8428427 TI - Multicentre guideline study. PMID- 8428428 TI - Stereotactic localization mammography. PMID- 8428429 TI - Long-term dobutamine therapy. PMID- 8428430 TI - Allergic bronchopulmonary aspergillosis. AB - The history, clinical features, diagnosis, treatment, and monitoring of allergic bronchopulmonary aspergillosis (ABPA) are described. ABPA is characterized by an immunologic reaction to the antigens of noninvasive Aspergillus fumigatus, which colonizes the bronchial lumen of affected individuals. It is a potential and often unrecognized pulmonary fibrotic complication in patients with asthma and cystic fibrosis. ABPA can occur at any age and may lead to cor pulmonale, respiratory failure, and death as a result of end-stage fibrotic lung destruction. Early diagnosis is essential for management of ABPA. Criteria for diagnosis of ABPA are (1) episodic bronchial obstruction, (2) peripheral blood eosinophilia, (3) immediate cutaneous reactivity to A. fumigatus, (4) precipitating serum antibodies to A. fumigatus, (5) elevated total serum IgE, (6) history of pulmonary infiltrates, (7) elevated serum IgE and serum IgG to A. fumigatus, and (8) proximal bronchiectasis. The total serum IgE concentration and chest roentgenograms can be used to monitor drug therapy. ABPA has five stages: acute, remission, exacerbation, corticosteroid-dependent asthma, and fibrotic. The most effective treatment for ABPA is oral prednisone 0.5 mg/kg/day for 14 days, on alternate days for three months, and tapering by 5 mg every two weeks for an additional three months. ABPA is a potentially fatal, noninfectious, inflammatory pulmonary disease coexistent with asthma and cystic fibrosis. With early diagnosis, adequate oral corticosteroid treatment, and IgE and chest roentgenographic monitoring, fibrotic lung complications can be averted and corticosteroid adverse effects minimized. PMID- 8428431 TI - FDA user fees approved. PMID- 8428432 TI - Helicobacter pylori and peptic ulcer disease. AB - The role played by Helicobacter pylori in the pathogenesis of peptic ulcer disease (PUD) is discussed, and the epidemiology, identification, diagnosis, eradication, and treatment of H. pylori infection are reviewed. Isolation of H. pylori from up to 100% of patients with duodenal ulcer and 80% of patients with gastric ulcer establishes a strong association between H. pylori and idiopathic PUD, although other factors also may be essential for the development of PUD. Invasive procedures for diagnosis of H. pylori infection include upper endoscopy and biopsy of gastroduodenal tissues followed by culture or the rapid urea test; noninvasive tests include the urea breath tests and serology. Although H. pylori is susceptible to a number of antimicrobials, eradication (as opposed to suppression) of this organism has been a major challenge. The most important predictive factor for clinical and microbiological efficacy is the pretreatment susceptibility of H. pylori to nitroimidazoles. Triple therapy with bismuth, metronidazole, and either amoxicillin or tetracycline has resulted in better clinical and microbiological outcomes than either monotherapy or dual therapy. Possible adverse effects of this regimen include nausea, vomiting, taste disturbance, and diarrhea. Anti-H. pylori therapy should be reserved for those patients who have recurrent symptomatic or intractable PUD. Currently, the regimen of choice includes bismuth, metronidazole, and either amoxicillin or tetracycline given for at least two weeks. PMID- 8428433 TI - Current recommendations of the joint national high blood pressure committee. AB - A great deal of new information has become available in the field of hypertension since the JNC report of 1988. The JNC V report has changed the categorization of blood pressure, modified suggested drugs for initial therapy, and recommended that diuretics or beta blockers be considered the first-line drugs of choice. Information concerning the J curve and end-stage renal disease has made therapeutic goals more challenging. One of the most important additions to this report is the new information on treating elderly patients, which had been lacking until last year. The report calls on pharmacists to assist with detecting, evaluating, and referring hypertensive patients. Pharmacists must take a leadership role in promoting compliance with antihypertensive therapy and can assist other health-care professionals by suggesting therapeutic alternatives to improve efficacy, reduce the frequency of administration, and lower costs. The complete JNC V report is an essential reference for the files of any pharmacist who is responsible for the care of hypertensive patients. PMID- 8428434 TI - FDA proposes more stringent pediatric-use labeling of prescription drug products. PMID- 8428435 TI - Teniposide granted marketing approval. PMID- 8428436 TI - Criteria for use of ticlopidine in adult inpatients and outpatients. PMID- 8428437 TI - Should patch testing be restricted to dermatologists only? PMID- 8428438 TI - Contact hypersensitivity to hydrocortisone-free-alcohol in patients with allergic patch test reactions to tixocortol pivalate. AB - It has been suggested that contact allergy to hydrocortisone alcohol is a frequent phenomenon. A recent study showed that all patients with allergic patch reactions to tixocortol pivalate reacted to intradermal hydrocortisone sodium phosphate. We studied patients with positive patch test reactions to tixocortol pivalate but negative to hydrocortisone alcohol, with penetration enhancers in hydrocortisone alcohol patch tests and oral challenges with hydrocortisone alcohol. Additionally, prick and intradermal tests with hydrocortisone sodium succinate were used. Using penetration enhancers and oral challenges enabled detection of more contact allergies to hydrocortisone alcohol compared to conventional patch testing alone. 9/12 patients with allergic reactions to tixocortol pivalate reacted to intradermal hydrocortisone sodium succinate. No immediate reactions were seen in prick or intradermal tests, suggesting that hydrocortisone contact hypersensitivity is probably not associated with immediate allergy to hydrocortisone. The present study suggests that allergic patch test reactions to tixocortol pivalate are caused by hypersensitivity to hydrocortisone alcohol itself or to one of its metabolites in the skin. PMID- 8428439 TI - Nickel, cobalt and chromium in consumer products: a role in allergic contact dermatitis? AB - In spite of the improved awareness of the potential for nickel, cobalt and chromium to cause skin allergy, the incidence of sensitization to them is generally on the increase, especially for nickel. We review data from the literature and industry on transition metal contamination of consumer products and assess the hazard to man. Consumer products are defined as personal care items and detergent/cleaning products used regularly in domestic work. The analytical data demonstrate that consumer products are a relatively minor source of contact with nickel, cobalt or chromium. The traces found in consumer products will not be the primary cause of sensitization to these metals: levels will be too low and exposure too brief. A person sensitized to these metals has many other more significant sources of daily contact such as earrings, jewelry and metal objects. It is therefore necessary to focus on decreasing the high exposure to these transition metals from other sources rather than on possible trace amounts found in consumer products. Current good manufacturing practice ensures that trace nickel, cobalt and chromium concentrations in consumer products are less than 5 ppm of each metal. It is recommended that this be accepted as a standard for maximum concentrations and that the target should be to achieve concentrations as low as 1 ppm. PMID- 8428440 TI - Occupational allergic contact dermatitis from mercury. AB - Occupational allergic contact dermatitis from metallic mercury is rare. Here we present the only 2 patients with relevant occupational mercury allergy detected at our clinic since 1974. The first patient was a dental nurse who became sensitized to metallic mercury from amalgam when handling uncured amalgam without protective gloves. The second patient had previously been sensitized to mercury from topical medicaments and developed work-related dermatitis when a mercury thermometer was broken at her place of work. Both patients had a positive patch test reaction to metallic mercury. PMID- 8428441 TI - Stability of the mercaptobenzothiazole compounds. AB - An analytical quantitative high-pressure liquid chromatography (HPLC) method was developed for simultaneous determination of all mercaptobenzothiazole derivatives in the mercapto mix patch testing standard. The stability of the mercaptobenzothiazoles constituting the mercapto mix was studied both in petrolatum and in buffer solution at pH 6.5, with and without glutathione. In petrolatum vehicle, dibenzothiazyl disulfide was the dominant compound found in stored mercapto mix. In buffer solution at pH 6.5, 2-mercaptobenzothiazole and the sulfenamide derivatives morpholinyl mercaptobenzothiazole and N-cyclohexyl-2 benzothiazyl sulfenamide were converted into dibenzothiazyl disulfide. In the presence of glutathione, both the sulfenamide derivatives and the dibenzothiazyl disulfide were rapidly converted into 2-mercaptobenzothiazole. The findings explain the "cross-sensitivities" reported for the mercaptobenzothiazole group as a result of chemical reactions resulting in one main hapten. The use of a single substance for patch testing for mercaptobenzothiazole hypersensitivity is proposed. PMID- 8428442 TI - Iatrogenic benign lymphoplasia induced by allergic contact dermatitis from squaric acid dibutylester: immunohistologic study of cellular infiltrates. AB - We report of a 62-year-old male patient with a dull red itchy nodule on the induction area of allergic contact dermatitis to squaric acid dibutylester, which had been used for the therapy of alopecia universalis. The excised biopsy specimen showed dense infiltration of lymphoid cells in the dermis and subcutaneous tissue, associated with the formation of lymphoid follicles. Immunohistologic analysis of the infiltrates indicated mixed proliferation of T- and B-cells. A biopsy specimen from the challenge area showed spongiosis in the epidermis and lymphoid cell infiltration in the upper dermis, while the infiltrates consisted mainly of T-cells. The following points are discussed: (i) the lesion had an iatrogenic origin and the causative agent was quite evident; (ii) the route of allergen application was only through the epidermis and not directly in the dermis; (iii) lymphoid cell infiltrates of the induction and challenge areas were different. PMID- 8428443 TI - Contact allergy to ophthalmic dipivalyl epinephrine hydrochloride: demonstration by patch testing. AB - A patient presented with a 3-month history of conjunctivitis and periocular eczema. He had a 3-year history of glaucoma and was being treated with 3 different locally applied eyedrops (Timoptic (timolol maleate) ophthalmic solution 0.25%, Pilocar (pilocarpine) ophthalmic solution 1%, and Propine (dipivalyl epinephrine hydrochloride) ophthalmic solution 0.1%). Patch testing with all 3 undiluted eyedrop solutions revealed a reaction only to Propine eyedrops. Patch testing to the individual components of Propine eyedrops revealed an allergic reaction to 0.5% dipivalyl epinephrine hydrochloride that was apparent on the 2nd patch test reading. After discontinuing the Propine eyedrops, the conjunctivitis as well as the periocular dermatitis resolved, proving that the positive patch tests were relevant to both. It was reported that re-exposure to dipivalyl epinephrine hydrochloride by intra-ocular challenge was necessary to diagnose this allergy. This is the first demonstration of dipivalyl epinephrine hydrochloride allergy by patch testing. Since the 1st patch test reading (2 days) was weak, 2nd patch test (4 days) readings may be important in diagnosing this allergy by patch testing. PMID- 8428444 TI - Allergic contact dermatitis from the herbicide Alachlor. PMID- 8428445 TI - Contact dermatitis from pramoxine. PMID- 8428446 TI - A case of "contact allergy to corticosteroid.". PMID- 8428447 TI - Allergic contact dermatitis from resin-reinforced plaster. PMID- 8428448 TI - Sensitization to levobunolol eyedrops. PMID- 8428449 TI - Edematous allergic contact cheilitis from a toothpaste. PMID- 8428450 TI - Bullae formation secondary to the use of a topical corticosteroid aerosol preparation. PMID- 8428451 TI - Fixed drug eruption from piroxicam. PMID- 8428452 TI - Occupational airborne contact urticaria due to xylene. PMID- 8428453 TI - Sensitivity to topical medicaments among suspected cases of footwear dermatitis. PMID- 8428454 TI - Allergic contact dermatitis due to an amino-substituted diacrylate in a UV-cured lacquer. PMID- 8428455 TI - Protein contact dermatitis from calf placenta extracts. PMID- 8428457 TI - Angioneurotic oedema following piperazine ingestion in an ethylenediamine sensitive subject. PMID- 8428456 TI - Contact sensitivity to nystatin in Timodine. PMID- 8428459 TI - Mild heat shock therapy of allergic contact reactions. PMID- 8428458 TI - Occupational allergic contact dermatitis due to 3-dimethylaminopropylamine. PMID- 8428460 TI - 1,2-benzisothiazolin-3-one. PMID- 8428461 TI - Time course of occlusive effects on skin evaluated by measurement of transepidermal water loss (TEWL). Including patch tests with sodium lauryl sulphate and water. AB - The time course for transepidermal water loss (TEWL) for a period of 3 h after removal of occlusive patch tests with sodium lauryl sulphate (SLS), water and empty chambers was studied in healthy volunteers. Patches were applied to the upper arm for 24 h. TEWL was measured immediately after removal of the patches, and every 30 min up to 3 h. For SLS and water patches, TEWL remained significantly increased for 3 h, as compared to normal adjacent skin, while for empty chamber patches, TEWL was only significantly increased for 30 min. A significant decrease from 0 to 30 min and from 30 to 60 min was observed for all patches, and for water patches, a significant decrease in TEWL was found from 60 to 180 min, while SLS patches remained constant. The prolonged increase in TEWL observed after SLS exposure is a well-known occurrence. The prolonged increase in TEWL after exposure to water is interpreted as transient damage to the water barrier of the skin. PMID- 8428462 TI - Nosocomial respiratory failure or iatrogenic ventilator dependency. PMID- 8428463 TI - High-frequency oscillatory ventilation in pediatric critical care. PMID- 8428464 TI - New study prepares the Society of Critical Care Medicine to face healthcare reform. PMID- 8428465 TI - Tension pneumothorax: diagnostic and therapeutic pitfalls. PMID- 8428466 TI - Lidocaine and acute coronary care. PMID- 8428467 TI - Imposed work of breathing and methods of triggering a demand-flow, continuous positive airway pressure system. AB - OBJECTIVES: To compare the inspiratory imposed work of breathing during spontaneous ventilation with continuous positive airway pressure using three methods of triggering "ON" the demand-flow system of a ventilator: a) conventional pressure triggering with the pressure measuring/triggering site inside the ventilator on the exhalation limb of the breathing circuit; b) tracheal pressure triggering from the tracheal or carinal end of the endotracheal tube; and c) flow-by (flow triggered) triggering. DESIGN: Multitrial tests under simulated clinical conditions using a mechanical lung model. SETTING: A research laboratory at a university medical center. INTERVENTIONS: Spontaneous breathing with continuous positive airway pressure, at peak sinusoidal inspiratory flow rate demands of 30, 60, and 90 L/min with sizes 6, 7, 8, and 9 mm internal diameter endotracheal tubes at each flow rate during conventional pressure triggering, tracheal pressure triggering, and flow-by. MEASUREMENTS AND MAIN RESULTS: Pressures were measured at the tracheal end of the endotracheal tube, "Y" piece of the breathing circuit, and inside the ventilator on the exhalation limb of the breathing circuit. Volume measured between the endotracheal tube and lung model and pressure measured at the tracheal end of the endotracheal tube were integrated to generate pressure-volume (work) loops to calculate the inspiratory imposed work of the total breathing apparatus (i.e., endotracheal tube, breathing circuit, and ventilator). Significantly (p < .05) greater decreases in pressure during spontaneous inhalation were measured for all methods of triggering at the tracheal end of the endotracheal tube than at the Y piece or inside the ventilator. Inspiratory-imposed work was significantly lower during tracheal pressure triggering compared with conventional pressure triggering and flow-by under most conditions. For example, with a 7-mm internal diameter endotracheal tube at a peak inspiratory flow rate demand of 60 L/min, imposed work was 382% and 315% lower, respectively, during tracheal pressure triggering compared with the conventional pressure triggering and flow-by triggering methods. Under all conditions, inspiratory imposed work was lower during flow-by triggering compared with conventional pressure triggering. The smaller the internal diameter of the endotracheal tube and the greater the peak inspiratory flow rate demand, the greater the inspiratory imposed work of breathing for all methods of triggering. Under all conditions, inspiratory-imposed work was significantly greater at a peak inspiratory flow rate demand of 90 L/min than at 60 L/min, and at a peak inspiratory flow rate demand of 60 L/min than at 30 L/min. CONCLUSIONS: An endotracheal tube is a resistor in the breathing apparatus over which a pressure decrease must be developed by the patient in order to inhale spontaneously. An endotracheal tube, therefore, imposes substantial resistance and work. The results indicate that the pressure measuring/triggering site for a ventilator's demand-flow system should be at the tracheal or carinal end of an endotracheal tube so as to effectively decrease the resistance of the endotracheal tube, thus, decreasing the patient's work of breathing. PMID- 8428468 TI - Detrimental effects of high-dose methylprednisolone sodium succinate on serum concentrations of hepatic and renal function indicators in severe sepsis and septic shock. The Methylprednisolone Severe Sepsis Study Group. AB - OBJECTIVE: To evaluate the effects of high-dose methylprednisolone sodium succinate on biochemical markers of hepatic and renal function in patients with severe sepsis and septic shock. DESIGN: Retrospective analysis of serial serum chemistries in 382 patients who were entered prospectively into a randomized, placebo-controlled, double-blind clinical trial of high-dose methylprednisolone or placebo in the sepsis syndrome. SETTING: The original study was conducted at 19 academic centers. PATIENTS: Adult patients in severe sepsis or septic shock who met the study entry criteria, which included a clinically defined source of infection and signs of systemic sepsis, were enrolled into the study. Three hundred eighty-two patients were evaluated. INTERVENTIONS: Patients received either methylprednisolone (30 mg/kg) or placebo by iv infusion every 6 hrs for four doses. Hemodynamic variables and serum concentrations of creatinine, urea nitrogen, bilirubin, and aspartate aminotransferase (AST) were recorded on entering the study, at 12 and 24 hrs, and at 3, 7, and 14 days after the first infusion of methylprednisolone or placebo. These data were analyzed retrospectively. MAIN OUTCOME MEASUREMENTS: Hemodynamic and biochemical data were analyzed to determine whether or not hepatic and renal function in the sepsis syndrome had been influenced by methylprednisolone treatment. RESULTS: Differences between methylprednisolone and placebo in hemodynamic variables, the occurrence rate of shock and recovery from shock, mortality rates and serum concentrations of creatinine and AST were not statistically significant. At 12 and 24 hrs, and at 3 and 7 days after the first drug infusion (of methylprednisolone or placebo), blood urea nitrogen was increased from baseline values in a significantly (p < .01) greater proportion of the methylprednisolone treated patients compared with placebo-treated patients. The frequency of increased serum bilirubin concentrations was significantly (p < .01) greater among methylprednisolone patients vs. the placebo group at 12 and 24 hrs. CONCLUSIONS: The frequency of acutely increased blood urea nitrogen and bilirubin concentrations in severe sepsis was increased significantly with high-dose methylprednisolone therapy. Similar frequencies of circulatory shock in the study groups excluded differences in global perfusion as a cause of this phenomenon. Possible adverse effects of pharmacologic concentrations of methylprednisolone in critically ill patients should be considered in planning treatment. PMID- 8428469 TI - Human sepsis increases lymphocyte intracellular calcium. AB - OBJECTIVE: To determine whether free intracellular calcium is increased during human bacterial sepsis. DESIGN: Prospective controlled study of lymphocyte free intracellular calcium concentrations from patients with sepsis compared with critically ill nonseptic patients and healthy subjects. SETTING: A large multidisciplinary ICU of a university hospital. PATIENTS: Eleven patients with sepsis, six patients after cardiac surgery, six patients with head injury, and 22 healthy control subjects. INTERVENTIONS: Blood samples obtained for lymphocyte isolation and measurement of free intracellular calcium concentrations. MEASUREMENTS: Lymphocytes were isolated using Ficoll-paque centrifugation and free intracellular calcium concentrations were measured using the fluorescent dye fura-2. We also evaluated the effect of septic serum, endotoxin, tumor necrosis factor (TNF), and lysophosphatidylcholine on lymphocyte free intracellular calcium concentrations. MAIN RESULTS: Mean (+/- SEM) lymphocyte free intracellular calcium concentrations were significantly (p < .05) higher in the septic patients (176 +/- 12 nM) compared with cardiac surgical (112 +/- 9 nM), head-injured (110 +/- 11 nM), or healthy control patients (112 +/- 5 nM). Endotoxin (0.1 and 1.0 mg/mL) and TNF (10 and 100 ng/mL) did not alter lymphocyte free intracellular calcium values. Lysophosphatidylcholine (100 and 200 microM) significantly increased lymphocyte free intracellular calcium in a dose-dependent manner. Septic serum had no effect on resting lymphocyte free intracellular calcium concentrations but potentiated the free intracellular calcium response to lysophosphatidylcholine. CONCLUSIONS: Lymphocyte intracellular calcium homeostasis is altered during human sepsis. In addition, circulating factors present in septic serum are capable of altering cellular calcium handling. PMID- 8428470 TI - Association of hypomagnesemia and mortality in acutely ill medical patients. AB - OBJECTIVE: To test the hypothesis that the mortality rate of acutely ill patients admitted to a medical ward or medical ICU is higher for those patients who present with hypomagnesemia than for those patients who do not present with hypomagnesemia. DESIGN: Prospective, observational study. SETTING: Emergency Department admissions to the medical ward and medical ICU of a tertiary care teaching hospital serving an inner city patient population. SUBJECTS: A total of 381 consecutive acutely ill patients. MEASUREMENTS: Serum magnesium concentrations and other metabolic variables were measured on admission from the Emergency Department. Acute Physiology and Chronic Health Evaluation (APACHE II) scores were computed for all patients, and mortality rates were determined for hypomagnesemic and normomagnesemic groups. MAIN RESULTS: Hypomagnesemic and normomagnesemic groups had comparable APACHE II scores and other variables. However, the mortality rates of the hypomagnesemic ward and medical ICU groups were approximately twice (p < .01) the rate of the normomagnesemic groups. Additionally, the duration of hospital survival in those patients who died was approximately 8 days less for hypomagnesemia than normomagnesemia, but not for ward admissions. Other associated metabolic abnormalities were frequently observed in both hypomagnesemic and normomagnesemic groups, including hypokalemia and hypocalcemia. CONCLUSIONS: Hypomagnesemia detected at the time of admission of acutely ill medical patients is associated with an increased mortality rate for both ward and medical ICU patients. PMID- 8428471 TI - Association between gastric intramucosal pH and splanchnic endotoxin, antibody to endotoxin, and tumor necrosis factor-alpha concentrations in patients undergoing cardiopulmonary bypass. AB - OBJECTIVES: To determine the association between gastric intramucosal pH, a minimally invasive marker reflecting the adequacy of oxygen delivery to the gastrointestinal tract, and splanchnic endotoxin, antibody to endotoxin, and tumor necrosis factor (TNF)-alpha concentrations in patients undergoing cardiopulmonary bypass. DESIGN: Single-arm, prospective study. SETTING: University hospital. PATIENTS: Adults (n = 10) free of hepatic, pulmonary, and renal disease undergoing nonemergent coronary artery bypass surgery. INTERVENTIONS: After induction of general anesthesia and endotracheal intubation, a tonometer nasogastric tube was positioned in the stomach, and triple-lumen fiberoptic catheters were inserted into the hepatic vein and pulmonary artery. Hepatic venous and mixed venous blood samples were analyzed for endotoxin, antibody to endotoxin, and TNF-alpha at six times: 30 mins after induction of anesthesia (time 1); during vena caval cannulation (time 2); after 15 mins of hypothermic cardiopulmonary bypass (time 3); during spontaneous left ventricular ejection after release of the aortic cross-clamp, but before termination of cardiopulmonary bypass (time 4); 15 mins after termination of cardiopulmonary bypass (time 5); and 1 hr after termination of cardiopulmonary bypass (time 6). Gastric intramucosal pH, systemic oxygen delivery (DO2), mixed venous oxygen saturation, hepatic venous oxygen saturation, and hepatic venous lactate concentrations were recorded at these same times. Data for each variable were compared with baseline values (time 1) for statistical significance. MEASUREMENTS AND MAIN RESULTS: Cardiopulmonary bypass was associated with an increase (p < .05) in systemic endotoxin concentrations from ventricular ejection until the end of the study. Virtually identical changes in the splanchnic circulation at this time approached, but did not reach, statistical significance, because hepatic venous endotoxin concentrations were higher than the mixed venous endotoxin concentrations at baseline (41.6 +/- 11.2 vs. 16.9 +/- 4.9 pg/mL). Gastric intramucosal pH was abnormal (< 7.35) at 15 mins (p > .05) and at 1 hr after termination of cardiopulmonary bypass (p > .05). The relationship between endotoxin and gastric intramucosal pH was not statistically significant (p = .15). The decrease in endotoxin antibody was small and statistically insignificant. TNF-alpha was not detected in any patient. Systemic DO2 decreased (p < .05) after 15 mins of hypothermic cardiopulmonary bypass, but returned to baseline values thereafter. There were no significant changes in mixed venous and hepatic venous oxygen saturation values. Splanchnic lactate concentrations increased at cannulation (p < .05), after 15 mins of hypothermic cardiopulmonary bypass (p < .05), and 15 mins after termination of cardiopulmonary bypass (p < .05). CONCLUSIONS: These observations are consistent with the hypothesis that impaired gut-barrier function is responsible for endotoxemia occurring during cardiopulmonary bypass. It is unclear whether increased mucosal permeability and mucosal acidosis are causally related phenomena or simply independent markers of damage to gut epithelium. PMID- 8428472 TI - Unreliability of blood pressure and heart rate to evaluate cardiac output in emergency resuscitation and critical illness. AB - OBJECTIVE: To evaluate the reliability of the vital signs to evaluate circulatory stability as reflected by cardiac index. DESIGN: Descriptive analysis based on data gathered prospectively, using a predetermined protocol. SETTING: University run county hospital, with a large trauma service. PATIENTS: Sixty-one high-risk trauma patients with accidental injury who were studied immediately after admission to the Emergency Department, and subsequently, 163 critically ill postoperative ICU patients. INTERVENTIONS: Standard fluid therapy, usually crystalloids, but occasionally packed red cell transfusions and colloids, as indicated by clinical criteria. MEASUREMENTS AND RESULTS: Arterial BP was measured by pressure transducer and arterial catheter; heart rate (HR) was measured by electrocardiograph signal, and cardiac output was measured by thermodilution. In sudden severe hypovolemic hypotension, the mean arterial pressure (MAP) nadir (lowest) roughly correlated (r2 = .25) with flow, but there was poor correlation (r2 = .0001) when all pressure and flow values were evaluated. The pressure and flow values were obtained throughout the course of the hypotensive episodes during the initial resuscitation in ICU patients and during terminal illnesses. CONCLUSIONS: Observations at the time of acute severe hypotensive crises that show rough correlation of MAP and cardiac index should not be extrapolated throughout the entire hypotensive period or to other less extreme clinical situations. The stress response to hypovolemia, with endogenous catecholamines and neural mechanisms, tends to maintain arterial pressure in the face of decreasing flow for a variable period of time. However, when these mechanisms are overwhelmed by prolonged hypovolemia, the pressure decreases precipitously, but not synchronously, with flow. We conclude that blood flow cannot reliably be inferred from arterial pressure and heart rate measurements until extreme hypotension occurs. PMID- 8428473 TI - Effect of intermittent pneumatic leg compression on intracranial pressure in brain-injured patients. AB - OBJECTIVE: To evaluate the effect of intermittent pneumatic leg compression on intracranial pressure and cerebral perfusion pressure in brain-injured patients. DESIGN: Prospective, sequential patient study. SETTING: Surgical/trauma ICU of a community hospital providing regional trauma care. PATIENTS: Twenty-four adult, brain-injured patients (mean Glasgow Coma Scale score = 6) who required hemodynamic and intracranial pressure monitoring. INTERVENTIONS: Placement of intermittent sequential pneumatic leg compression devices for prevention of venous thrombosis. MEASUREMENTS: Mean arterial pressure (MAP), heart rate, central venous pressure, and intracranial pressure were measured at baseline, and at 0, 10, 20, and 30 mins of intermittent pneumatic leg compression. Cerebral perfusion pressure was calculated as the difference between MAP and intracranial pressure. RESULTS: No significant changes in MAP, central venous pressure, or intracranial pressure occurred during the study interval. Calculated cerebral perfusion pressure remained unchanged. A total of 23 of 24 study patients had intracranial pressure controlled by hyperventilation or pharmacologic measures within the normal range at the time of study. CONCLUSION: Intermittent pneumatic leg compression results in no significant changes in intracranial pressure or cerebral perfusion pressure in stable, brain-injured patients who have intracranial pressure controlled by medical means. PMID- 8428474 TI - Autonomic cardiovascular state after severe brain injury and brain death in children. AB - OBJECTIVE: To study and compare the autonomic cardiovascular state of children after severe brain injury and brain death. DESIGN: Prospective clinical study. SETTING: Pediatric ICU. PATIENTS: Pediatric patients suffering severe brain injury caused by trauma, anoxia, or hemorrhage. INTERVENTION: None. MEASUREMENTS AND MAIN RESULTS: We analyzed cardiorespiratory parameters, heart rate power spectra, plasma catecholamine concentrations, and the response to the cold pressor test in nine brain-dead patients and compared the results with the test findings of 11 patients with severe brain injury. Low-frequency total heart rate power (p < .03), peak amplitude (p < .02), and plasma catecholamine concentrations (p < .001) were different with no overlap of values between groups. Cold pressor testing in patients with severe brain injury showed changes in respiratory rate and low-frequency heart rate power that were +/- 20% to 100% from baseline values; however, there were no measurable changes in brain-dead patients. CONCLUSIONS: Our results support the concept of a damaged sympathetic cardiovascular system in severe brain injury and complete interruption of the autonomic cardiovascular pathways in brain death. Since determination of brain death may be difficult, our findings have implications for corroborating brain death using autonomic cardiovascular testing. PMID- 8428475 TI - Preoperative intensive care unit consultations: accurate and effective. AB - OBJECTIVES: To determine if a structured preoperative ICU consultation would correctly assign patients to preoperative invasive monitoring, postoperative ICU care, or recovery room care, and to compare morbidity, mortality, and resource utilization among all groups. DESIGN: Prospective, observational study. SETTING: A university hospital. PATIENTS: A total of 475 patients who were referred preoperatively by surgeons for ICU consultation and were evaluated by ICU physicians. INTERVENTIONS: Patients assessed to have clinical evidence of cardiovascular compromise were admitted preoperatively to the ICU for invasive hemodynamic monitoring and optimization. Patients without such evidence, but who were to undergo major operations or had anticipated major fluid replacement were independently selected for invasive monitoring by anesthesiologists. Patients who developed physiologic instability or became unstable due to hemorrhage also underwent invasive monitoring. Nonmonitored patients who remained stable were given postoperative ICU care or went to the recovery room based on an assessment by the surgeon and anesthesiologist at the end of the operation. MEASUREMENTS AND MAIN RESULTS: Of 8,916 elective surgical cases, ICU physicians were consulted in 475 (5.3%) patients preoperatively. Sixty-seven patients were admitted preoperatively to the ICU for invasive hemodynamic monitoring and optimization; 60 patients had surgery (0.7% of elective cases, 12.6% of ICU consultations). Patients selected for ICU preoperative monitoring were older than non-monitored patients and had higher numbers of cardiovascular and total risk factors than any other group. They had higher Acute Physiology and Chronic Health Evaluation (APACHE II) scores, higher Therapeutic Intervention Scoring System (TISS) points, a higher number of complications, and longer ICU stays than non-monitored postoperative ICU patients. In addition, they had a higher number of complications than nonmonitored recovery room patients. APACHE II scores, TISS points, number of complications, and ICU days in the preoperative ICU admission group were not increased when compared with all other monitored patients. Neither hospital days nor total hospital charges were increased when compared with the other elective ICU patients. Patients selected for ICU preoperative monitoring who underwent surgery had an 11.7% mortality rate and accounted for four of five cardiovascular-related deaths. CONCLUSIONS: A small number of high-risk patients can be selected for preoperative monitoring on the basis of clinical assessment without increasing ICU stay or hospital bills. A structured preoperative consultation correctly identifies those patients who need monitoring and ICU care, but does not overutilize scarce and expensive ICU beds. PMID- 8428476 TI - Increased serum concentrations of procollagen type III peptide in severely injured patients: an indicator of fibrosing activity? AB - OBJECTIVES: To determine the serum concentrations of procollagen type III peptide in severely injured patients with different outcomes and to evaluate the relationship between serum procollagen type III peptide concentrations, sources of increased posttraumatic fibrotic activity (wounds, lung, liver, kidney), and decreased elimination of procollagen type III peptide (liver). DESIGN: Prospective study. SETTING: Surgical ICU, university hospital. PATIENTS: Fifty seven patients (mean injury severity score: 38.5 points, range 13 to 75 points), between 16 and 70 yrs of age, treated in our institution within 6 hrs after the accident. MEASUREMENTS: Serial measurements were started on admission and continued on a 6-hr basis. After 48 hrs, the monitoring interval was extended to 24 hrs until recovery (but at least until day 14) or death. At each point of evaluation, pulmonary and circulatory function parameters and chest radiographs (once a day) were evaluated, the results were recorded, and blood samples were drawn to determine procollagen type III peptide, total bilirubin, creatinine, gamma-glutamyl transferase, polymorphonuclear elastase, and other parameters. Statistic evaluation was done with the Wilcoxon test, Spearman rank correlation, and a multiple regression model. RESULTS: Mean procollagen type III peptide serum concentrations (+/- SD) were significantly different in patients who died (8.0 +/ 3.8 U/mL) compared with those patients who survived with organ failure (2.7 +/- 1.3 U/mL) or without complications (1.4 +/- 0.5 U/mL), respectively. Significant correlations of procollagen type III peptide concentrations with the serum bilirubin concentrations (r = .7), days with need of mechanical ventilation (r = .64), PaO2/FIO2 ratio (r = -.6), polymorphonuclear elastase (r = .6), serum creatinine concentrations (r = .55), and injury severity score (r = .33) were observed. There was a tendency toward higher serum procollagen type III peptide concentrations in patients with severe skeletal injuries. CONCLUSIONS: Serum procollagen type III peptide concentrations in severely injured patients may be considerably increased in correlation with injury severity and outcome. Procollagen type III peptide serum concentrations seem to reflect the sum of increased collagen formation from wound healing and fibrogenesis of mediator related organ damage (especially lung) and decreased procollagen type III peptide excretion due to impaired liver function. Further data are necessary to evaluate the role of hepatic elimination in these patients. PMID- 8428477 TI - Physiologic response of stress and aminoglycoside clearance in critically ill patients. AB - OBJECTIVE: To examine the relationships between aminoglycoside clearance and physiologic parameters associated with the physiologic response to injury. DESIGN: Cross-sectional study of surgical patients receiving aminoglycoside pharmacokinetic monitoring and parenteral nutritional support. SETTING: An adult surgical ICU. PATIENTS: Fifty-four surgical/trauma patients who had Gram-negative sepsis. INTERVENTIONS: Measurements of the physiologic stress response to injury were associated with aminoglycoside clearance in 54 surgical/trauma patients who had Gram-negative sepsis. Measurements used to estimate the magnitude of the stress response included a 24-hr urinary urea nitrogen excretion, blood urea nitrogen, peak temperature, serum albumin, bilirubin, and transferrin concentrations. MEASUREMENTS AND MAIN RESULTS: Mean drug clearance rate (4.4 +/- 2.5 [SD] L/hr) was related to the physiologic measurements using correlation and regression techniques. Collectively, all physiologic indices (utilized) explained 59% of the variance in drug clearance (p < .001), an amount similar to the variance explained by creatinine clearance alone (53%). When all six physiologic measurements were included into a multiple regression model that included creatinine clearance, the total variance explained increased to 73%. CONCLUSIONS: Along with renal function estimates, the physiologic response to stress should be considered when treating critically ill patients with aminoglycosides and other, similar, renally eliminated drugs. PMID- 8428478 TI - Use of polyethylene glycol-bound superoxide dismutase, polyethylene glycol-bound catalase, and nimodipine to prevent hypoxic ischemic injury to the brain of newborn pigs. AB - OBJECTIVE: To determine if polyethylene glycol-bound superoxide dismutase and catalase and nimodipine, alone or in combination, will ameliorate hypoxic ischemic injury to the brain. SUBJECTS: A total of 78 newborn (0 to 3 days) pigs were used. DESIGN: Prospective, blinded, randomized, controlled trial. INTERVENTIONS: The piglets were subjected to hypoxic ischemic brain injury. Carotid arteries were ligated at time 0 and BP was reduced one third by hemorrhage. At 15 mins, FIO2 was reduced to 0.6. At 30 mins, carotids were released, blood was reinfused, and FIO2 was increased to 1.0. Pigs were randomly assigned at time 35 mins to receive either: 10,000 U/kg of polyethylene glycol bound superoxide dismutase and catalase (group 1); 0.5 mg/kg of nimodipine (group 2); both 10,000 U/kg of superoxide dismutase and catalase and 0.5 mg/kg of nimodipine (group 3); or no drugs (controls). MEASUREMENTS: The time after reoxygenation for return of electroencephalogram, respiration, blink and pain were recorded in minutes as well as a neurologic examination at 1, 2, and 3 days and pathologic examination of the brain at 3 days, both by blinded observers. MAIN RESULTS: There were no significant differences in the four groups. CONCLUSIONS: Polyethylene glycol-bound superoxide dismutase and catalase, and nimodipine, either alone or in combination, do not ameliorate hypoxic ischemic injury to the brain in the newborn pig when given 5 mins after reoxygenation. PMID- 8428479 TI - Histamine-2-receptor blockade does not affect hyperemia after cerebral ischemia in young swine. AB - OBJECTIVE: To determine whether histamine-2 (H2) receptor antagonists can blunt the increase in cerebral blood flow after cerebral ischemia. DESIGN: Prospective, randomized trial. SETTING: Animal laboratory. SUBJECTS: Piglets aged 1 to 2 wks. INTERVENTIONS: Three groups of piglets were anesthetized with pentobarbital, mechanically ventilated, and hemodynamically monitored. Each animal was randomized to receive one of three treatments 20 mins before ischemia: famotidine (0.6 mg/kg i.v.), cimetidine (5 mg/kg i.v.), or an equivalent volume of saline (3 mL). Ischemia was maintained for 15 mins by tightening ligatures around the brachiocephalic trunk and left subclavian arteries plus the induction of hemorrhagic hypotension (50 mm Hg). Reperfusion was initiated by the reinfusion of shed blood and removal of the ligatures. MEASUREMENTS AND MAIN RESULTS: The three groups were not different with regard to baseline arterial blood gases, cerebral perfusion pressure, microsphere-determined cerebral blood flow, or oxygen consumption (VO2). Ischemia (cerebral blood flow < 5 mL/min/100 g of tissue) was demonstrated in all brain regions for all groups during arterial ligation with hemorrhage-induced hypotension. Blood flow rates were increased compared with preischemic values in all regions at 7 mins of reperfusion. For example, mean forebrain blood flow rates increased from 36 +/- 3 to 95 +/- 14 (SEM) mL/min/100 g of tissue in the group receiving saline, from 36 +/- 3 to 102 +/- 9 mL/min/100 g of tissue in the group receiving famotidine, and from 31 +/- 4 to 119 +/- 26 mL/min/100 g in the group receiving cimetidine. Overall, the three groups did not differ in regard to blood flow rates and cerebral VO2 values during ischemia and at 30 mins of reperfusion. CONCLUSIONS: We demonstrated that famotidine or cimetidine as compared with saline did not affect the pattern or extent of hyperemia after ischemia. This finding does not support the hypothesis that a H2-receptor mechanism is necessary for postischemic hyperemia. PMID- 8428480 TI - Acute respiratory failure neuropathy: a variant of critical illness polyneuropathy. AB - OBJECTIVE: To describe a severe axonal polyneuropathy that follows acute respiratory failure and, in turn, causes continued ventilator dependence and paralysis after resolution of the primary illness. DESIGN: Retrospective chart review of three patients and prospective analysis of two patients. SETTING: Respiratory and neurologic ICUs of a general hospital. PATIENTS: Five critically ill patients after an episode of acute respiratory failure. Neuromuscular blocking drugs were used in four patients, intermittently in two patients, high doses of corticosteroids were briefly administered in four, four patients had multiple organ failure, three patients had sepsis, but weakness preceded these complications in two patients. INTERVENTIONS: None. MAIN RESULTS: All patients had moderate-to-severe limb weakness with reduced or absent reflexes. Sensation was relatively preserved and the spinal fluid protein concentrations were normal. Electrophysiologic studies showed a severe, acute axonal motor neuropathy in four patients and normal studies in the fifth that later demonstrated denervation. Sensory potentials were mildly or not affected. Two quadriparetic patients died at 2.5 months, one remained weak and ventilator dependent several months after onset, and two patients recovered to walk in 4 to 6 months. CONCLUSIONS: Severe axonal motor neuropathy after acute respiratory failure probably represents a variant of "critical illness polyneuropathy" that can be recognized from the temporal course of a conversion from primarily pulmonary to a pattern of neuromuscular ventilatory failure. PMID- 8428481 TI - High-frequency oscillatory ventilation in pediatric respiratory failure. AB - OBJECTIVE: To evaluate the safety and effectiveness of high-frequency oscillatory ventilation using a protocol designed to achieve and maintain optimal lung volume in pediatric patients with respiratory failure. SETTING: Tertiary care pediatric ICU in a university hospital. DESIGN: A prospective, clinical study. PATIENTS: Seven patients aged 1 month to 15 yrs with diffuse alveolar disease and airleak with a variety of primary diagnoses, including pneumonia, adult respiratory distress syndrome, and pulmonary hemorrhage. INTERVENTIONS: After varying periods of conventional mechanical ventilation (16 to 216 hrs), patients were managed with high-frequency oscillatory ventilation using a "high-volume" strategy that consisted of incremental increases in mean airway pressure and lung volume to achieve an arterial oxygen saturation of > or = 90%, with an FIO2 of < or = 0.6. MEASUREMENTS AND MAIN RESULTS: Ventilatory settings, including FIO2 and mean airway pressure, hemodynamic parameters (cardiac index, systemic and pulmonary vascular resistance indices, oxygen delivery [DO2] and oxygen extraction ratio) and the oxygenation index (oxygenation index = [FIO2 x mean airway pressure x 100]/PaO2) were monitored during the transition to high-frequency oscillation and throughout the course of the high-frequency oscillatory ventilation with rapid and sustained reductions in mean airway pressure (p = .0001, repeated-measures analysis of variance [ANOVA]) and a trend toward decreasing oxygenation index (p = .08, repeated-measures ANOVA). In the four patients from whom hemodynamic data were obtained, there were no compromises of cardiac index or DO2 despite a significant increase in mean airway pressure (26 +/- 2 to 35 +/- 2 cm H2O) during conversion from conventional ventilation to high-frequency oscillation. CONCLUSIONS: High-frequency oscillatory ventilation, using a high-volume strategy, may be used safely and effectively in pediatric patients with respiratory failure and with high predicted mortality rates. High mean airway pressure during oscillatory ventilation does not appear to compromise DO2. Whether this technique can alter morbidity or mortality rates in this population awaits prospective randomized study. PMID- 8428482 TI - Descriptive analysis of critical care units in the United States: patient characteristics and intensive care unit utilization. AB - OBJECTIVE: To gather data about occupancy, admission characteristics, patients' ages, and types of therapy utilized in ICUs in the United States. DESIGN AND SETTING: Survey instruments were mailed to the administrators of 4,233 hospitals to gather information from the medical director of the institutions' respective ICUs for the purpose of developing a database on ICUs in the United States. The sampling frame for this study was based on all American Hospital Association (AHA) hospitals stating they had ICUs. MEASUREMENTS: Census questionnaires solicited information on occupancy, where the patients were admitted from, length of stay, therapies rendered, intensive care diagnoses, and resuscitation status, as well as other information. MAIN RESULTS: Data were obtained regarding 32,850 ICU beds, with 25,871 patients from 2,876 separate ICUs in 1,706 hospitals in the United States. The census response rate was 40% of the AHA hospitals that stated they had ICUs, with specific ICU data on 38.7% of the nation's ICUs. Overall, the responding units reported a mean occupancy rate of 84% of total bed capacity and 87% of available beds. As hospital size increased, so did ICU occupancy. Nearly 17% of all of the critical care patients had been in the units for > 14 days. More precisely, 49% of all responding units indicated that they had one or more "chronic" (> 14-day length of stay) patients. Most patients were admitted to the units from the emergency room (38%), operating room/postanesthesia care unit (22%), and the general hospital floor (16%). Neonatal units were exceptions to this observation, where most patients came from the delivery room (60%). Admission from other hospitals represented a significantly larger group of patients in the cardiac care, pediatric, and neonatal units. Respondents indicated that many of their current patients were elderly, with 43% of these patients aged 65 to 84 yrs and with 4% being > or = 85 yrs of age. The 47% of patients > or = 65 yrs of age increased to 58% when the neonatal and pediatric units were eliminated from the analyses. For all units responding to the survey, the leading primary admitting intensive care diagnoses were postoperative management, ischemic heart disorder, respiratory insufficiency/failure, and prematurity. Elimination of units predominantly treating children (pediatric and neonatal) from the analysis left "adult" units with three primary admitting diagnoses: ischemic heart disease, postoperative management, and respiratory insufficiency/failure with variation according to specific unit type. The leading diagnoses in pediatric units were respiratory insufficiency/failure, postoperative management, and congenital abnormalities. For neonatal units, prematurity was the primary admitting diagnosis, accounting for 59% of these units' patients. Respondents reported 5.3 +/- 10.9% of patients had received cardiopulmonary resuscitation (CPR) before admission into the critical care unit. Only 6.0 +/- 11.9% of patients in these critical care units had instructions that CPR not be performed while in the unit. CONCLUSIONS: This report should be viewed as the beginning step of an effort to improve both the information base available on critical care medicine and the performance of ICUs. Our survey findings provide an introduction into the everyday workings of critical care units throughout the United States. Research is required to determine which patients will benefit from intensive care and how to efficiently utilize the vast technology we have available for them in a world with limited financial resources, an aging population, and a multiplicity of societal and ethical concerns. PMID- 8428483 TI - Guidelines for granting privileges for the performance of procedures in critically ill patients. Guidelines Committee; Society of Critical Care Medicine. PMID- 8428484 TI - Hemodynamic measurements during a tension pneumothorax. PMID- 8428485 TI - Neonatal pneumomediastinum: indications for, and complication of, treatment. PMID- 8428486 TI - Acute valproic acid intoxication: enhanced drug clearance with oral-activated charcoal. PMID- 8428487 TI - Successful treatment of acute myocarditis using extracorporeal membrane oxygenation. PMID- 8428488 TI - Symposium: Tissue Perfusion in Critical Illnesses. PMID- 8428489 TI - Multiple organ failure: the pilgrim's progress. PMID- 8428490 TI - Associations between intramucosal acidosis in the gut and organ failure. AB - OBJECTIVE: To alert health professionals to the need for early detection and prevention of shock in critically ill patients. By describing the associations between intramucosal acidosis in the gut and multiple system organ failure, the author demonstrates how noninvasive measurement of gut intramucosal pH can be used to monitor the adequacy of tissue oxygenation in the splanchnic organs and predict splanchnic ischemia within minutes of its onset. DATA SOURCES: Review and analysis of current medical literature on shock and organ failure, combined with the author's prior research and expertise in the areas of tissue oxygenation and tonometric monitoring in the critically ill. CONCLUSIONS: The presence of defective tissue oxygenation in splanchnic organs and in gut ischemia may be detected within minutes of its occurrence via measurements of intramucosal pH in the gut. Measurement of intramucosal pH, obtained noninvasively with an intraluminally located gastrointestinal tonometer, provides an absolute metabolic measure of the adequacy of mucosal oxygenation. The putative consequences of intramucosal acidosis and associated mucosal injury include nosocomial pneumonia, myocardial depression, sepsis from enteric organisms, multiple system organ failure, and death. Through the use of routine monitoring of the adequacy of gut mucosal oxygenation, ischemic mucosal injury and its putative consequences can be prevented, resulting in reduced frequency of multiple organ failure and improved outcome. PMID- 8428491 TI - Systemic mediators released from the gut in critical illness. AB - OBJECTIVE: To discuss the mediators released from the gut in critical states, with emphasis on the intestinal mucosal barrier function, mediators of bacterial origin, and myocardial depressant factors. DATA SOURCES: Relevant articles that have been published in the English language literature. STUDY SELECTION: No special study has been carried out for the present discussion. DATA EXTRACTION: Information from the literature has been used to illustrate important points in the discussion. DATA SYNTHESIS: Due to decreased mucosal blood flow, increased short-circuiting of oxygen in the mucosal countercurrent exchanger, and increased oxygen demand in sepsis mucosal injury develops rapidly in the gut after various forms of shock and splanchnic ischemia. In addition, due to increased generation of oxygen-derived radicals, injury may also occur with reperfusion. As a consequence of increased permeability of the intestinal mucosal barrier between the luminal content and the sterile interior milieu, increased translocation of bacteria and bacterial endotoxin occurs. In addition, cardiodepressant factors are released, as is evident from in vivo and in vitro studies. No such factor has been fully identified chemically. CONCLUSIONS: Intestinal mucosal injury, as seen in critical illness, may induce increased translocation of bacteria and endotoxin and release of myocardial depressant factors into the circulation. PMID- 8428492 TI - Endotoxin-induced organ injury. AB - OBJECTIVE: To review the effects of endotoxemia on the major organ systems of the body and discuss potential mechanisms of tissue injury. DESIGN: Appraisal of 60 articles representing a cross section of studies relating to in vivo and in vitro responses to endotoxin. MAIN METHODS: Cell cultures, isolated tissue preparations, animal and human studies. RESULTS: Endotoxemia results in the activation of numerous cellular and hematogenous mediators. These mediators range from prostaglandins, thromboxanes, and leukotrienes, to complement components. Tumor necrosis factor may be responsible for initiating many of the observed responses to endotoxin. Species and tissue specificity are a prominent feature of the response to endotoxin. CONCLUSIONS: No single agent can yet be implicated as the common mediator of endotoxin-induced organ injury. Endotoxin initiates the elaboration of a cascade of secondary mediators that amplify the response to the initial insult. The relative importance of individual agents as mediators of the response to endotoxin varies with the experimental model studied. PMID- 8428493 TI - Goals for the resuscitation of shock. AB - OBJECTIVE: To remind practitioners of the conventional goals of resuscitation of overt hypotensive or uncompensated shock ("ABC," for airway, breathing, and circulation) and to introduce additional goals, represented by successive letters of the alphabet, to aid clinicians in recognizing the persistence of compensated shock in the splanchnic organs and in achieving more complete resuscitation by eliminating inadequate tissue perfusion in these organs. DATA SOURCES: Review and analysis of current medical literature on shock and organ failure, combined with the author's prior research and expertise in the areas of tissue oxygenation and tonometric monitoring in the critically ill. CONCLUSIONS: In traumatic and septic shock, multiple system organ failure is associated with a persistent state of compensated shock in which hypotension and oliguria are corrected, but in which inadequate perfusion persists in the splanchnic organs and especially in the mucosal lining of the gut. The additional goals recommended include "D" for increasing the delivery of oxygen to levels that meet the metabolic demand by all tissues in the body, especially those tissues within the splanchnic circulation, and "E" for ensuring extraction and utilization of oxygen by the tissues. Future needs for goals that address reperfusion injury, translocation of bacterial toxins, and the release of toxic mediators are also considered. PMID- 8428494 TI - Adoptive immunotherapy of gram-negative sepsis: use of monoclonal antibodies to lipopolysaccharide. AB - OBJECTIVE: To provide a succinct overview of the scientific rationale for adoptive immunotherapy of Gram-negative sepsis using antibodies directed at epitopes in the core region of the lipopolysaccharide molecule. DATA SOURCES: Relevant, English-language primary and secondary (i.e., review) articles dealing with the structure or toxicity of lipopolysaccharide or adoptive anti-endotoxin immunotherapy in animals or man. STUDY SELECTION AND DATA EXTRACTION: The review emphasizes the findings obtained in several recent multicenter, randomized, prospective trials of monoclonal antibodies to core determinants of lipopolysaccharide. DATA SYNTHESIS: Lipopolysaccharide, a component of the outer cell wall of Gram-negative bacteria, is a complex molecule consisting of three regions: lipid A, core polysaccharide, and O-antigenic side chains. Epitopes in the lipid A and core polysaccharide regions are highly conserved across species and strains of Gram-negative bacteria. Because of the availability of mutant strains of bacteria that synthesize lipopolysaccharides lacking O-specific side chains and, in some instances, portions of the core polysaccharide moiety, investigators have been able to develop polyclonal and monoclonal antibodies to core-region determinants. In experimental animals, many of these antibodies have been shown to protect against the deleterious effects of either purified heterologous lipopolysaccharides or viable heterologous Gram-negative bacteria. Recently, two different monoclonal "anti-core" antibodies (HA-1A and E5) were evaluated in prospective, placebo-controlled, double-blind, randomized trials. In the pivotal trial of HA-1A, a human monoclonal IgM, no treatment effect was discernible when data from all patients enrolled in the study were analyzed. However, in a subset of patients with positive blood cultures for Gram-negative organisms, there was a decrease in all-cause mortality over a 28-day observation period in patients treated with HA-1A, as compared with those patients treated with placebo (p = .014). Interpretation of this finding, however, is confounded because multiple other subsets and end-points also were analyzed, necessitating an adjustment in the p value necessary to reject the null hypothesis. A murine monoclonal IgM called E5 was evaluated in two separate trials. In the first trial, improved survival over a 30-day observation period was observed in a subset of patients with Gram-negative sepsis without refractory shock (p = .01). However, in a second trial of E5, which focused on this subset, a statistically significant improvement in survival was not observed in patients with documented Gram-negative sepsis without shock. CONCLUSIONS: Adoptive immunotherapy using monoclonal anticore antibodies seems to improve survival rate in selected patients with Gram-negative sepsis. Nevertheless, because of concerns about costs and the interpretation of the results from the completed clinical trials, these new agents have generated enormous controversy. The precise role of adoptive immunotherapy against lipopolysaccharide in the practice of critical care medicine in the United States remains unclear. PMID- 8428495 TI - Adequacy of gut oxygenation in endotoxemia and sepsis. AB - OBJECTIVE: To provide a succinct overview of current notions regarding sepsis induced alterations in mesenteric perfusion and oxygen transport. DATA SOURCES: Selected English-language articles dealing with mesenteric perfusion and gut mucosal function during sepsis or endotoxicosis in experimental animals or man. STUDY SELECTION AND DATA EXTRACTION: The review emphasizes findings obtained using a well-characterized porcine model of acute, resuscitated endotoxicosis. Other experimental and clinical studies are discussed as well. DATA SYNTHESIS: Total hepatosplanchnic perfusion and oxygen uptake are increased in most patients with compensated sepsis. No data are currently available from clinical studies regarding the effect of sepsis on mesenteric perfusion per se. Data are unavailable regarding either total hepatosplanchnic or mesenteric blood flow in patients with decompensated sepsis (i.e., septic shock). Therefore, current ideas regarding mesenteric perfusion in sepsis derive primarily from studies using animal models. In a normodynamic porcine endotoxicosis model, mesenteric perfusion and oxygen delivery (DO2) are markedly decreased. The changes in flow and DO2 are accompanied by intestinal mucosal acidosis and increased permeability to hydrophilic solutes, suggesting that these latter phenomena are a consequence of lipopolysaccharide-induced mesenteric hypoperfusion. This idea is supported by the observation that maintenance of normal mesenteric blood flow ameliorates gut mucosal acidosis and hyperpermeability in endotoxic pigs. However, because transmesenteric oxygen consumption is unchanged in endotoxic pigs, the precise mechanistic relationship between hypoperfusion and altered barrier function remains puzzling. CONCLUSION: Mesenteric hypoperfusion may be an important factor leading to alterations in gut epithelial permeability in endotoxicosis and sepsis. PMID- 8428496 TI - Adequacy of tissue oxygenation. AB - OBJECTIVE: This paper reviews presently available techniques for monitoring the adequacy of tissue oxygenation, emphasizing the practical and theoretical problems that exist with presently used measurements. DATA SOURCES: The data are based on a review of the literature. DATA SYNTHESIS: Most of the presently available techniques focus on global measurements of oxygen transport and utilization that may be insensitive to changes occurring in vital tissues critically important to overall homeostasis. As a result, they are often insensitive, nonspecific, and become abnormal only at a very late stage of disease. CONCLUSIONS: In attempting to develop tools to assess adequate tissue oxygenation, emphasis should be placed on the monitoring of individual tissues that are felt to be highly susceptible to reduced oxygen delivery and key to overall survival. Preliminary data involving measurements of the interstitial pH of the gastrointestinal tract suggest that this measurement may be one approach to pursue. PMID- 8428497 TI - Tissue oxygenation in low flow states and during hypoxemia. AB - BACKGROUND AND METHODS: Tissue oxygenation during ischemia and hypoxia is critical to cellular viability. Adequate tissue oxygenation reflects a balance between oxygen delivery (DO2) and oxygen demand. At critical levels of DO2, oxygen consumption (VO2) declines due to supply dependency, indicating inadequate tissue oxygenation and impaired cellular metabolism. The mechanism by which DO2 is compromised (hypoxia vs. ischemia) determines the level of delivery at which VO2 becomes supply dependent. Tonometry is a method used to assess the adequacy of gut oxygenation. RESULTS: Studies using tonometric monitoring of intramural pH have shown that tonometry detects early tissue anaerobiosis and net adenosine triphosphate (ATP) hydrolysis. A decrease in intramural pH strongly correlates with the onset of supply dependency in tissue VO2. Inadequate intestinal oxygenation determined by tonometry has predicted clinical outcomes and complications in patients at risk. CONCLUSIONS: Gut tonometry may provide an early indication of inadequate tissue oxygenation. This early recognition is necessary to mitigate adverse consequences, including ATP catabolism, the production of reactive oxygen metabolites, and the activation of the inflammatory process, which can lead to progressive cellular dysfunction and cell death. PMID- 8428498 TI - Bacterial translocation after thermal injury. AB - OBJECTIVES: To review the mechanisms responsible for bacterial translocation after thermal injury. Areas investigated were the rate of bacterial translocation, blood flow to the gastrointestinal tract, potential of reversibility of mesenteric vasoconstriction, specific vasomediators responsible for postburn mesenteric vasoconstriction, potential reversal of gut mucosal atrophy with decreased translocation, and evidence of gut mucosal damage after thermal injury. DESIGN: Using three different animal models consisting of rats, sheep, and minipigs, the objectives were defined. Using the sheep model, the relationship of decreased mesenteric blood flow after thermal injury was defined along with rates of translocation, and the potential reversibility of the postburn mesenteric vasoconstriction and its effect on translocation. The effect of smoke inhalation and the combination of thermal injury and inhalation injury on rates of translocation are explained. Using minipigs, the role that thromboxane A2 plays on the postburn mesenteric vasoconstriction was defined by blocking thromboxane A2 synthesis with OKY046, a specific thromboxane synthetase inhibitor. Evidence of gut mucosal injury was determined using ornithine decarboxylase as an indicator of gut mucosal damage and subsequent repair in the minipig model. The rat model was used to demonstrate gut mucosal atrophy after thermal injury and the potential for reversal of atrophy with the use of bombesin, a specific gut mucosal growth stimulator. RESULTS: After thermal injury, there were significant decreases in mesenteric blood flow. There was also an increase in bacterial translocation. Selective infusion of nitroprusside into the cephalic mesenteric artery prevented the post-burn mesenteric vasoconstriction and attenuated bacterial translocation. Smoke inhalation and smoke inhalation with thermal injury resulted in mesenteric vasoconstriction and increased rates of bacterial translocation. OKY046 infusion prevented the postburn increase in mesenteric vascular resistance. There were increased concentrations of ornithine decarboxylase within the colonic mucosa, indicating a previous injury and the presence of ongoing repair. Likewise, there was gut mucosal atrophy after thermal injury with bacterial translocation. Treating with bombesin attenuates the postburn mucosal atrophy and prevents bacterial translocation. CONCLUSIONS: Thermal injury is associated with mesenteric vasoconstriction. This postburn mesenteric vasoconstriction results in damage to gut mucosa and allows for increases in bacterial translocation. The postburn mesenteric ischemia can be ameliorated with nitroprusside infusion, thus preventing translocation. Thromboxane A2 appears to be a major mediator of the postburn decrease in mesenteric blood flow. Likewise, prevention of postburn gut mucosal atrophy with bombesin attenuates bacterial translocation. PMID- 8428499 TI - Vasoactive mediators and splanchnic perfusion. AB - OBJECTIVE: To provide an overview of the splanchnic hemodynamic response to circulatory shock. DATA SOURCES: Previous studies performed in our own laboratory, as well as a computer-assisted search of the English language literature (MEDLINE, 1966 to 1991), followed by a selective review of pertinent articles. STUDY SELECTION: Studies were selected that demonstrated relevance to the splanchnic hemodynamic response to circulatory shock, either by investigating the pathophysiology or documenting the sequelae. Article selection included clinical studies as well as studies in appropriate animal models. DATA EXTRACTION: Pertinent data were abstracted from the cited articles. RESULTS OF DATA SYNTHESIS: The splanchnic hemodynamic response to circulatory shock is characterized by a selective vasoconstriction of the mesenteric vasculature mediated largely by the renin-angiotensin axis. This vasospasm, while providing a natural selective advantage to the organism in mild-to-moderate shock (preserving relative perfusion of the heart, kidneys, and brain), may, in more severe shock, cause consequent loss of the gut epithelial barrier, or even hemorrhagic gastritis, ischemic colitis, or ischemic hepatitis. From a physiologic standpoint, nonpulsatile cardiopulmonary bypass, a controlled form of circulatory shock, has been found experimentally to significantly increase circulating levels of angiotensin II, the hormone responsible for this selective splanchnic vasoconstriction. CONCLUSIONS: While angiotensin II has been viewed primarily as the mediator responsible for the increased total vascular resistance seen during (and after) cardiopulmonary bypass, it may also cause the disproportionate decrease in mesenteric perfusion, as measured in human subjects by intraluminal gastric tonometry and galactose clearance by the liver, as well as the consequent development of the multiple organ failure syndrome seen in 1% to 5% of patients after cardiac surgery. PMID- 8428500 TI - Therapeutic potential of intraluminal oxygenation. AB - OBJECTIVE: To discuss the potential therapeutic value of intraluminal oxygenation of the gut in critical illness. DATA SOURCES: Relevant articles published in the English language literature. STUDY SELECTION: No special study has been carried out for the present presentation. DATA EXTRACTION: Information from the literature has been used to illustrate a potential way of preventing multiple organ failure. DATA SYNTHESIS: Animal experiments have demonstrated that oxygenation of the intestinal mucosa by providing gaseous oxygen to the mucosa or by perfusing the gut lumen with oxygenated fluids may prevent the development of mucosal injury that otherwise occurs in ischemic states. All available experience is based on experiments, and thus far, the clinical relevance of this observation is not tested. CONCLUSIONS: Intraluminal oxygenation may prevent the development of intestinal mucosal injury that is otherwise seen in conditions causing intestinal ischemia. PMID- 8428501 TI - Hepatic blood flow during cardiopulmonary bypass. AB - OBJECTIVES: To establish the effects of different modes of cardiopulmonary bypass on hepatic blood flow, with the aim of increasing understanding of the hemodynamic factors that may lead to hepatic dysfunction in patients after cardiopulmonary bypass. The anatomical and physiologic characteristics that are unique to the hepatic circulation are also reviewed, together with an account of the known specific effects on the liver of several hemodynamic stimuli that are commonly present during cardiopulmonary bypass. DATA SOURCES: The entire world literature on the subjects of clinical and experimental cardiopulmonary bypass, extracorporeal circulation, blood gas tensions, hypothermia, hypotension, and hepatic blood flow was searched via Index Medicus, up to and including 1991. DATA EXTRACTION: Data are presented only from those studies that employed sound cardiopulmonary bypass or blood flow measurement technique. For investigations on hypothermia, data for temperatures < 28 degrees C are not included. Details of one recent experimental study in the dog by the author are highlighted, in which the relative effects of pump flow rate, temperature, and type of perfusion (pulsatile or nonpulsatile) are presented; the data are critically compared and contrasted with those data obtained by earlier workers. DATA SYNTHESIS: Hepatic hemodynamics are characterized by a dual supply of blood from the hepatic artery, actively controlled within the liver, and the portal vein, principally regulated by prehepatic resistance vessels. Portal flow may modulate the hepatic arterial circulation through the "hepatic arterial buffer response." Hypotension caused by hemorrhage causes a decrease in portal blood flow but hepatic arterial flow is maintained by autoregulation and by the "buffer" response. Hypothermia (28 degrees C) has little effect on hepatic arterial flow, but portal flow may increase. During cardiopulmonary bypass, total liver blood flow is better maintained at a pump flow rate of 2.4 than at 1.2 L/min/m2. Perfusion at 28 degrees C causes an increase in portal flow and a slight decrease in hepatic arterial flow. Total hepatic blood flow is better preserved during cardiopulmonary bypass at 1.2 L/min/m2 by pulsatile than by nonpulsatile flow; however, no significant difference was noted between pulsatile and nonpulsatile perfusion at a bypass flow rate of 2.4 L/min/m2, particularly at 28 degrees C. CONCLUSIONS: During cardiopulmonary bypass, hepatic blood flow is better maintained by high pump flow than by low pump flow rates. Hypothermic cardiopulmonary bypass may benefit the hepatic circulation, although the additional advantages usually gained by the use of pulsatile perfusion may be partly lost when hypothermia is combined with a high pump flow rate. PMID- 8428502 TI - Adequacy of tissue oxygenation in cardiac surgery: regional measurements. AB - OBJECTIVE: The aim of the study was to measure visceral and peripheral perfusion and oxygenation during and immediately after cardiac surgery. METHODS: Central hemodynamics, blood gases, core temperature, visceral and peripheral tissue perfusion, and oxygenation were studied in eight patients undergoing coronary artery bypass grafting and in the early postoperative period in another group of ten coronary artery bypass grafting patients. DESIGN: The variables were measured after induction of anesthesia, after sternotomy, during cardiopulmonary bypass, after rewarming and after closing the wounds. Postoperatively, the measurements were carried out hourly up to 8 hrs after the arrival of patients in the ICU. PATIENTS: The patients were operated on under moderate hemodilution and systemic hypothermia. The inspired oxygen concentration was maintained at 30 volumes percent during the postoperative study. MEASUREMENTS: Visceral perfusion was indirectly assessed by determining the gastric intramucosal pH. Peripheral tissue perfusion was assessed in the upper extremity by continuous recording of subcutaneous tissue PO2, laser-Doppler skin erythrocyte flux, transcutaneous PO2, and fingertip temperature. Transcutaneous PO2 index (transcutaneous PO2/PaO2) was calculated. MAIN RESULTS: Gastric intramucosal pH, PaO2, and transcutaneous PO2 reached maximum values during cardiopulmonary bypass at the deepest level of hypothermia, and gastric intramucosal pH reached its minimum at the end of the operation. During the first 3 hrs after admission of patients to the ICU, gastric intramucosal pH decreased progressively, reached its minimum value at 5 hrs, and increased slowly thereafter. Subcutaneous tissue PO2, laser-Doppler skin erythrocyte flux, and fingertip temperature decreased markedly during cardiopulmonary bypass, increased during rewarming, and decreased again at the end of surgery. The peripheral vascular bed was vasoconstricted on ICU admission, as indicated by the low values of subcutaneous tissue PO2, transcutaneous PO2, transcutaneous PO2 index, laser-Doppler skin erythrocyte flux, and fingertip temperature. These variables began to increase over the next 2 to 4 hrs and reached their maximum value by the end of the 8-hr postoperative study period, indicating complete vasodilation of the peripheral vascular bed. CONCLUSIONS: These data suggest that the visceral perfusion of patients is well maintained during cardiopulmonary bypass, while, at the same time, these patients develop hypoperfusion and hypoxia of peripheral tissues. After closing the wounds, gastric intramucosal pH, transcutaneous PO2 index (transcutaneous PO2/PaO2), and other peripheral tissue perfusion variables were at the lowest values, indicating impending residual hypothermia and tissue hypoperfusion after rewarming. During the first few hours in the ICU, both the visceral and peripheral oxygenation and perfusion variables reflected hypoperfusion of tissues coinciding in time with the period most vulnerable for hemodynamic disasters and cardiac arrhythmias. PMID- 8428503 TI - Splanchnic lactate production in cardiac surgery patients. AB - OBJECTIVE: To review the pathophysiology of lactic acidosis in patients undergoing open-heart surgery, with special reference to the splanchnic circulation. DATA SOURCE: MEDLINE search of pertinent experimental and clinical research studies. RESULTS: Lactate is an end-product of anaerobic metabolism and is in dynamic equilibrium with its precursor, pyruvate. The ratio of serum lactate to pyruvate concentrations in arterial blood is normally < or = 10:1. In patients with lactic acidemia, measurement of serum pyruvate concentrations may yield valuable clinical information. Lactate/pyruvate ratios > 10:1 suggest that oxygen delivery (DO2) is inadequate to meet metabolic demand, whereas increases in both lactate and pyruvate values with preservation of normal lactate/pyruvate ratios suggest a defect in oxidative utilization (e.g., a fractional increase in the inactive form of the pyruvate dehydrogenase enzyme complex) despite adequate DO2. Meaningful changes in regional oxygen kinetics occur during extracorporeal circulation. Increased production of endogenous vasoconstrictors in response to nonpulsatile flow, together with a decrease in arterial oxygen content resulting from the addition of 2 L of pump prime to the patient's circulation at the beginning of cardiopulmonary bypass, decrease DO2 to the gastrointestinal tract. The effect of this reduction is mitigated, in part, by lowering core temperature and reducing tissue oxygen demand. CONCLUSIONS: The abdominal organs tolerate reductions in DO2 when they are cold (25 degrees C), since gastric intramucosal pH (a marker of inadequate DO2), and hepatic venous lactate/pyruvate ratios and oxygen saturation during the first half of cardiopulmonary bypass are normal. As surgery nears completion and core temperature is increased, tissue oxygen demands escalate. The presence of gastric mucosal acidosis, coupled with lactic acidemia and oxygen desaturation of hepatic venous blood, suggest that delivery of oxygen to the abdominal organs at the conclusion of cardiopulmonary bypass is insufficient to meet demand. A growing proportion of cardiac surgery patients are older and many have concomitant medical problems that can impair their recovery. Useful strategies are needed to reduce the occurrence of splanchnic ischemia in these and other high-risk populations if surgical outcome is to improve in the future. PMID- 8428504 TI - Cytokines and growth factors in endothelial dysfunction. AB - OBJECTIVE: The purpose of this focused review is to call attention to the important interrelationships between growth factors (e.g., transforming growth factor-beta) and cytokines (e.g., tumor necrosis factor [TNF]) on endothelial function as characterized by the ability of the endothelium to release endothelium-derived relaxing factor. BACKGROUND AND METHODS: Myocardial ischemia followed by reperfusion leads to a severe degree of endothelial dysfunction characterized by an impaired vasodilator response to endothelial-dependent vasodilators. The reduction in endothelial-dependent responses occurs rapidly after the onset of reperfusion. Endothelial dysfunction was studied in coronary artery as well as superior mesenteric artery rings, and the effects of cytokines, transforming growth factor-beta, and free radicals were assessed. RESULTS: Oxygen derived free radicals (i.e., superoxide anion) and cytokines (i.e., TNF) contribute to this endothelial dysfunction, which is characterized by a loss of endothelium-derived relaxing factor, now known to be identical to nitric oxide. Agents such as superoxide dismutase or transforming growth factor-beta can significantly preserve endothelial integrity during ischemia-reperfusion states. CONCLUSIONS: Endothelial dysfunction is an early and critical phenomenon occurring in ischemia-reperfusion injury both in the coronary and the mesenteric circulations. PMID- 8428505 TI - Tissue perfusion in critical illnesses. Antioxidant therapy. AB - OBJECTIVES: To provide an overview of the mechanisms of free radical-mediated reperfusion injury and to review the available antioxidant agents and strategies available to prevent or ameliorate this injury. DATA SOURCES: Previous studies performed in our own laboratory, as well as a computer-assisted search of the English language literature (MEDLINE, 1966 to 1991), followed by a selective review of pertinent articles. STUDY SELECTION: Articles representing the mechanisms of free-radical generation, free-radical-mediated tissue injury, and antioxidant function. DATA EXTRACTION: Pertinent data were abstracted from the cited articles. RESULTS OF DATA SYNTHESIS: The small intestine is disproportionately susceptible to ischemia during circulatory shock. Ischemia activates xanthine oxidase, which then generates a massive burst of superoxide free radicals when oxygen is reintroduced at reperfusion. These radicals and their toxic oxygen metabolites, both by direct action and by the secondary activation of circulating neutrophils, then generate much of the mucosal injury that had been previously attributed to anoxia itself. CONCLUSIONS: Recent evidence indicates that although the mucosal epithelial cell is rich in xanthine oxidase, it is probably endothelial cell xanthine oxidase that triggers this mucosal reperfusion injury, not only in the intestine, but in other organs as well. Antioxidant therapies directed toward the ablation of free-radical generation at reperfusion offer a potential therapy for these injuries and their sequelae. PMID- 8428506 TI - Opportunistic pneumonia. AB - Patients with abnormalities in their immune defense mechanisms are frequently encountered in the practice of medicine. These patients have increased susceptibility to infections that often involve the lung. Many of these infections are caused by opportunistic organisms that typically do not produce disease in individuals with a normally functioning immune system. To complicate the evaluation of these patients, they frequently develop noninfectious lung disease, which can have a radiographic appearance that is similar to infection. The purpose of this article is to present an approach to the evaluation of chest radiographic abnormalities in the immunocompromised patient with suspected pneumonia. Clinical information that is pertinent in the evaluation of immunocompromised patients is discussed. The various chest radiographic patterns seen in both infectious and noninfectious diseases that occur in the immunocompromised patient are reviewed. Integration of clinical information and radiographic findings in the development of a list of differential diagnoses is stressed. PMID- 8428507 TI - Fluorescence in situ hybridization mapping of the cystic fibrosis transmembrane conductance regulator (CFTR) gene to 7q31.3. AB - We have used the fluorescence in situ hybridization (FISH) technique to refine the localization of the cystic fibrosis transmembrane conductance regulator (CFTR) gene on human chromosome 7. The result shows that the gene is most likely located within band q31.3. PMID- 8428508 TI - Analysis of aphidicolin-induced chromosome fragility in the domestic pig (Sus scrofa). AB - Aphidicolin (APC)-sensitive fragile sites were identified in chromosome preparations from peripheral lymphocyte cultures of 12 four-way crossbred pigs. A chi 2 analysis demonstrated that aphidicolin-induced breakage events and previously reported in vivo chromosome rearrangement events were not independent. Comparison of expected Poisson and negative binomial distributions to observed breakage patterns indicated that the negative binomial distribution provided a better fit to experimental data. The negative binomial distribution is consistent with a distribution of breakage rates, i.e., non-constant rates of breakage at chromosomal loci across the genome. PMID- 8428509 TI - A balanced autosomal reciprocal translocation in an azoospermic bull. AB - This appears to be the first reported case of a bull with a balanced autosomal reciprocal translocation associated with azoospermia. The analysis includes somatic chromosome banding, conventional meiotic analysis, and electron microscopy of synaptonemal complexes (SCs). The karyotype of the bull was found to be 60,XY,rcp(8;13)(q11;q24). Electron microscopy of SCs in microspread pachytene spermatocytes revealed a high incidence of terminal asynapsis of the smallest arm of the quadrivalent. Most quadrivalents with such asynapsis and few with nonhomologous synapsis showed associations with the XY sex bivalent, leading to complete meiotic arrest at late pachynema. Except for one diakinetic cell, no diplotene and subsequent stages were encountered in air-dried meiotic preparations. The presence of degenerating primary spermatocytes in SC preparations, as well as in testicular sections, and the absence of spermatozoa in ejaculates confirm the chromosomally derived male sterility of the bull. X chromosome reactivation, evidenced by the cytomorphological reversal of associated sex bivalents, appeared to be the initial step in the degeneration of spermatocytes. Consequently, the formation of a separate, fully developed XY body, which was previously demonstrated on the periphery of spermatocyte nuclei in fertile bulls, could not be attained in this case. Extensive end-to-end association of autosomal bivalents in meiotically arrested, as well as degenerating, spermatocytes was a consistent and unique observation of this study. Such associations may lead to enhanced reactivation of the X chromosome. PMID- 8428510 TI - Synaptonemal complex analysis of a reciprocal translocation, rcp(20;24) (q17;q25), in a subfertile bull. AB - A reciprocal translocation was identified in a subfertile artificial insemination bull. Somatic chromosome investigation of G-banded metaphases demonstrated a 60,XY,rcp(20;24)(q17;q25) karyotype for the carrier. Synaptonemal complex analysis of the translocation by electron microscopy revealed an irregular pairing behavior of the chromosome axes involved, which resulted in a variety of configurations at pachytene. Not only was the expected quadrivalent configuration present, but also a trivalent plus univalent and two heteromorphic bivalents. Most common was an incompletely or completely paired quadrivalent configuration, which was non-homologously paired. XY association with the multivalent was seen only rarely. Histological analysis of testicular tissue showed meiotic arrest in some tubules. However, the semen picture was normal. PMID- 8428511 TI - The human endothelin-1 gene (EDN1) encoding a peptide with potent vasoactive properties maps distal to HLA on chromosome arm 6p in close linkage to D6S89. AB - We determined the precise genetic location of the human endothelin-1 gene (EDN1), which encodes a peptide with extremely potent vasoactive properties and is apparently involved in a spectrum of diseases ranging from hypertension to asthma. Analyzing the segregation of a four-allele EDN1 polymorphism in 40 CEPH families including 480 individuals, we detected significant linkage of EDN1 to DNA markers spanning the telomeric half of chromosome arm 6p. EDN1 was closest to the highly polymorphic nucleotide-repeat marker D6S89 at a theta = 0.06 with the highest pairwise LOD score Zmax = 31.2. Subsequent multipoint analysis placed EDN1 at 8 cM distal to D6S89; EDN1 was flanked at its telomeric site at a 13-cM distance by the gene encoding the A subunit of blood clotting factor XIII (F13A1). Furthermore, EDN1 was located at approximately 34-36 cM distal to the HLA region defined by HLA-A, -B, and -DRB1, and 31 cM proximal to the most telomeric marker D6S7. This location of EDN1 on the primary genetic map is strongly supported with odds of 2.7 x 10(12):1 against the next best alternative. PMID- 8428512 TI - Localization of the horse (Equus caballus) alpha-globin gene complex to chromosome 13 by fluorescence in situ hybridization. AB - The alpha-globin gene complex in Equus caballus has been mapped by fluorescence in situ hybridization to the telomeric region of the long arm of chromosome 13. This is the first equine gene to be mapped to this chromosome. PMID- 8428513 TI - A PCR-based method to amplify DNA with random primers: determining the chromosomal content of porcine flow-karyotype peaks by chromosome painting. AB - We present here a new PCR-based technique that allows the production of several micrograms of DNA from only 300 flow-sorted chromosomes. During the first two PCR cycles, the annealing temperature is decreased to 30 degrees C, and numerous random loci are amplified under nonspecific conditions. As demonstrated here for pig chromosomes 1 and 18, the PCR products may be used to identify the chromosomal content of the flow-karyotype peaks of any species by fluorescence in situ hybridization. PMID- 8428514 TI - Comparative analyses of heterochromatin in Microtus: sequence heterogeneity and localized expansion and contraction of satellite DNA arrays. AB - Southern blotting, C-banding, base-specific fluorochrome staining, and fluorescence in situ hybridization were used to analyze the constitutive heterochromatin in eight species and subspecies of arvicolid rodents (genus Microtus). Autosomal centromeric regions portrayed considerable variability between species in the amount of C-band-positive material present; e.g., M. chrotorrhinus showed relatively little, whereas M. cabrerae exhibited extensive centromeric staining. Autosomal interstitial C-bands were noted in M. guentheri and two subspecies of M. ochrogaster. All Y chromosomes examined were predominately or completely heterochromatic, as were substantial portions of the giant X chromosomes in three species (M. agrestis, M. cabrerae, and M. chrotorrhinus). Hoechst 33258 staining (with its affinity for AT binding sites) showed bright fluorescence on the heterochromatin of the sex chromosomes of M. agrestis and moderate fluorescence on those of M. cabrerae and M. chrotorrhinus; however, only the heterochromatin of M. cabrerae and M. chrotorrhinus hybridized with an AT-rich satellite DNA probe (MSAT-160) isolated from M. chrotorrhinus. Hoechst 33258-bright autosomal centromeres of M. arvalis and M. cabrerae also hybridized to the probe, whereas the Hoechst 33258-bright Y chromosomes of M. arvalis and M. guentheri did not. Two pairs of autosomes in M. guentheri are comprised of six distinct regions, based upon C-banding, Hoechst 33258 staining, chromomycin A3/distamycin A staining, and in situ hybridization. The centromeric regions of acrocentric autosomes known to retain conserved G-banding patterns may exhibit variable hybridization intensity when different species or subspecies are compared. M. ochrogaster portrays considerable intersubspecific variability in the size and location of autosomal telomeric and interstitial C-bands that are also sites of hybridization. These latter two findings illustrate that dramatic differences in copy number of the tandem satellite array can exist at homologous chromosomal positions both within and between species. PMID- 8428515 TI - New markers, D16FC1 and Tp12, differentiate between rat chromosomes 16 and 17. AB - Problems in differentiating rat chromosomes 16 and 17 cytogenetically can be resolved with unique probes mapped to these chromosomes. Using somatic cell hybridization and nonisotopic in situ hybridization, probes D16FC1 and Tp12 were localized to 16p16-->p15 and 17q12.1-->q12.2, respectively. The locations of these probes can serve as reference points to facilitate mapping of future probes to rat chromosomes 16 and 17. PMID- 8428516 TI - Partial sequence data from three evolutionarily conserved loci from the proximal short arm of the human X chromosome; assignment of DXF34S1 to Xp11.21-cen. AB - DNA sequence data have been obtained from three clones derived from the human X chromosome which contain evolutionarily conserved sequences. Primers have been designed which enable these loci to be defined as sequence-tagged-sites (STS's). The assignment of one of the loci, DXF34S1, has been refined to Xp11.21-cen, thus limiting the novel pericentromeric segment of homology defined by this locus to the extreme proximal region of Xp. PMID- 8428518 TI - A "hot spot" of recombination coincides with an interstitial telomeric sequence in the Armenian hamster. AB - The chromosomes of male Armenian hamsters (Cricetulus migratorius) exhibit highly localized terminal and interstitial chiasmata. An interstitial telomeric sequence detected by in situ hybridization was clearly visible at the primary constriction of a large metacentric in both mitotic and meiotic chromosomes. This telomeric sequence was the site of a chiasma in 69% of all 58 diakinesis/metaphase I cells examined. The frequency of meiotic exchange at this site was therefore 34.5%, an extraordinary "hot spot" of recombination. Subsequent chiasma interference can be assumed to reduce recombination and tighten linkage along both arms of male hamster chromosomes. These observations suggest that telomere-promoted recombination may represent a second pathway of recombination, mechanistically different from the more heterodispersed process of meiotic exchange. PMID- 8428517 TI - Isolation of a somatic cell hybrid retaining the der(16)t(12;16)(q13;p11.2) from a myxoid liposarcoma cell line. AB - We developed cell line LIS-3/SV40 from a primary myxoid liposarcoma with the t(12;16)(q13;p11.2) and trisomy 8. Following fusion of LIS-3/SV40 to mouse A9 cells, we obtained hybrid LIS-3/SV40/A9-B4 which contained the der(16) but neither the der(12) nor some of the normal human chromosomes, including 12 and 16. Furthermore, microclone library ML12q1315 was constructed from microdissected fragments of chromosome bands 12q13-15 and its microclones were found to span the breakpoint in LIS-3/SV40. PMID- 8428519 TI - The fibroblast growth factor receptor 3 gene (FGFR3) is assigned to human chromosome 4. AB - Fibroblast growth factor receptors (FGFRs) have recently been isolated and shown to be transmembrane tyrosine kinase receptors. The FGFR1 gene has previously been assigned to human chromosome 8 and the FGFR4 gene to human chromosome 5. Here we demonstrate, by using somatic cell hybrids, that the FGFR3 gene localizes to human chromosome 4, showing that it, too, resides on a chromosome distinct from those on which other FGFRs have been localized. PMID- 8428520 TI - The gamma fibrinogen gene (FGG) maps to chromosome 17 in both cattle and sheep. AB - The gamma fibrinogen gene (FGG) was localised in both cattle and sheep using in situ hybridisation. The probe employed was a 1-kb bovine cDNA fragment. Based on observations of QFQ-banded chromosome preparations, this locus is on bovine chromosome 17q12-->q13 and on the homologous sheep chromosome 17. This localisation is, to our knowledge, the first assignment to chromosome 17 in either the bovine or ovine genome. In addition to localising FGG to this chromosome, the assignment provisionally maps the previously unassigned syntenic group U23, containing (besides FGG) the genes for mitochondrial aldehyde dehydrogenase 2 (ALDH2), interleukin 2 (IL2), immunoglobulin lambda (IGL), and beta fibrinogen (FGB), to chromosome 17 in cattle and probably to the same chromosome in sheep. PMID- 8428521 TI - Assignment of the human slow skeletal muscle troponin gene (TNNI1) to 1q32 by fluorescence in situ hybridisation. AB - The human gene for slow-twitch skeletal muscle troponin I (TNNI1) has previously been mapped to 1q12-->qter using somatic cell hybrids. The TNNI1 locus has now been further localised to 1q32 using fluorescence in situ hybridization. This result confirms the previous assignment of this locus and maps the gene to a single chromosome band. PMID- 8428522 TI - Chromosome identity of human prostate cancer cell lines, PC-3 and PPC-1. PMID- 8428523 TI - Recovery of infectious HIV following the passage of apoptotic cellular debris through phagocytic macrophages. PMID- 8428524 TI - HIV infection of the lung. Role of virus-infected macrophages in the pathophysiology of pulmonary disease. PMID- 8428525 TI - Alveolar macrophages in HIV-1 infection express accessory molecules, activation markers, and release increased biological response modifiers. PMID- 8428526 TI - HIV impairs alveolar macrophage mannose receptor function against Pneumocystis carinii. PMID- 8428527 TI - Bronchoalveolar lavage fluid and handling of mycobacteria. PMID- 8428528 TI - CD8 cells mediate delayed hypersensitivity following intrapulmonary infection with Cryptococcus neoformans. PMID- 8428529 TI - Potential role of viruses in the pathogenesis of pulmonary fibrosis. PMID- 8428530 TI - Altered patterns of lung lymphocyte accumulation in silicosis in cytokine sufficient (C3H/HeN) and cytokine-deficient (C3H/HeJ-LPSd) mice. PMID- 8428531 TI - Altered production and regulation of monocyte chemoattractant protein-1 from pulmonary fibroblasts isolated from patients with idiopathic pulmonary fibrosis. PMID- 8428532 TI - Recombinant interferon-gamma reduces PDGF-induced lung fibroblast growth but stimulates PDGF production by alveolar macrophages in vitro. PMID- 8428533 TI - Tolerance to hapten prevents specific delayed type hypersensitivity and pulmonary interstitial fibrosis in the mouse model. PMID- 8428534 TI - The T cell and the airway's fibrotic response in asthma. PMID- 8428535 TI - A role for T helper-1 cells in the induction of airway hyperresponsiveness. PMID- 8428536 TI - Immunohistochemical characterization of T lymphocytes and eosinophils in the bronchial wall of actively sensitized guinea pigs. PMID- 8428537 TI - Changes in T lymphocyte subsets and activation following chronic antigen inhalations in monkeys. PMID- 8428538 TI - Lung allograft rejection: role of tumor necrosis factor-alpha and interleukin-6. PMID- 8428539 TI - Initiation and maintenance of the granulomatous response. PMID- 8428540 TI - Increased TNF-alpha and IL6 mRNA expression by alveolar macrophages in chronic beryllium disease. PMID- 8428541 TI - CD3+, CD4+, CD8-, Ia- T cells adoptively transfer murine experimental hypersensitivity pneumonitis. PMID- 8428542 TI - Hypersensitivity pneumonitis in the athymic nude mouse. Another evidence of T cell dependency. PMID- 8428543 TI - Thomas L Petty Aspen Lung Conference 35th Annual Meeting: Immunology of the Lung. PMID- 8428544 TI - T cell tolerance. Giles F. Filley Lecture. AB - T cell tolerance to self is induced by several mechanisms. Immature T cells may die, and mature T cells may die or be inactivated by contact with self. While we do not understand completely what governs which of these courses will be adopted by an autoreactive T cell, 2 points are clear. Self is in part distinguished from nonself by the fact that the former is always present, while the latter is only intermittently in the body. Secondly, it seems that the default response for T cells is "off" rather than "on." That is, peripheral T cells will become tolerant to antigen unless some other phenomenon such as adjuvant triggers them to respond. One other point should be mentioned. In spite of the power of the mechanisms which causes self tolerance, tolerance is not complete. Autoreactive T cells specific for cryptic antigens, expressed in the brain or eye for example, can easily be demonstrated. Many autoimmune diseases may be caused by aberrant activation and exposure to target organ of these cells, rather than failure of the normal mechanisms of tolerance themselves. PMID- 8428545 TI - T cell receptor variable beta-gene expression in the normal lung and in active pulmonary sarcoidosis. PMID- 8428546 TI - Adhesion molecules involved in leukocyte recruitment and lymphocyte recirculation. PMID- 8428547 TI - Characterization of the leukocyte integrin subunit beta 7. PMID- 8428548 TI - In situ regulation of pulmonary macrophage TNF-alpha mRNA expression by IL2. PMID- 8428549 TI - Pulmonary responses to antigen. PMID- 8428550 TI - Programmed cell death and apoptosis in lymphocyte development and function. PMID- 8428551 TI - [Reversible spinal cord compression caused by extramedullary hematopoietic foci in thalassemia]. AB - A now 42-year-old Thai woman was known to have been anaemic since childhood. When aged 33 years she was diagnosed as having beta zero/HBE thalassaemia. Computed tomography demonstrated a tumour in the posterior mediastinum, histologically found to be an extramedullary haematopoietic focus. Subcutaneous infusion of deferoxamine (2 g five times weekly), initiated because of massive iron overload, reduced the serum ferritin level from 3,460 ng/ml to less than 500 ng/ml. The haemoglobin level in the subsequent years was between 6 and 8 g/dl. Five years later sensory deficits were noted from the 5th thoracic vertebra downwards. Magnetic resonance imaging demonstrated a tumour which compressed the spinal cord: it, too, was an ectopic haematopoietic focus. The neurological symptoms disappeared after radiotherapy with 3,000 cGy, but they recurred 4 years later. Because of the low radiation reserve of the spinal cord, hypertransfusion treatment was initiated, namely 16 RBC concentrates within 4 months and afterwards two transfusions every 3 months. By this means the haemoglobin level was kept at about 9 g/dl. The tumour had regressed 4 months after onset of treatment. For 2 years since the beginning of the hypertransfusion treatment the patient has remained free of neurological symptoms. PMID- 8428552 TI - [Nuclear medicine methods for the detection of gastrointestinal hemorrhages]. PMID- 8428553 TI - [Drug-induced immune hemocytopenias]. PMID- 8428554 TI - [A central venous catheter in acute myocardial infarct?]. PMID- 8428555 TI - [beta-Sympathomimetics against hyperkalemia in kidney failure]. PMID- 8428556 TI - [Fluoride prophylaxis]. PMID- 8428557 TI - [Hypoparathyroidism following radiotherapy?]. PMID- 8428558 TI - [The therapy of Salmonella enteritis]. PMID- 8428559 TI - [Thyroid diseases following therapy for Hodgkin's disease]. PMID- 8428560 TI - [The risk factors of sudden infant death]. PMID- 8428561 TI - [The effect of alcohol on portal vein hemodynamics in nutritional-toxic liver cirrhosis]. AB - The influence of alcohol on portal vein haemodynamics was assessed prospectively in 30 patients (20 men, 10 women; mean age 54.3 [34-70] years) with nutritional toxic cirrhosis of the liver (Child-Pugh stages A-C) and portal vein hypertension. During the period of observation hepatic vein occlusion pressure as an indirect measure of portal vein pressure was repeatedly determined. In addition, the size of oesophageal varices and the Child-Pugh stage were monitored. After complete alcohol abstinence of one year, portal vein pressure fell from 23.11 to 12.43 mm Hg (-46%, P < 0.001), the Child-Pugh score from 8.08 to 7.2 (-10.9%, not significant), and the size of oesophageal varices was reduced from grade 1.33 to grade 0.79 (-40%, P < 0.02). On resuming alcohol abuse, portal vein pressure increased by an average of 10 mm Hg (+60%, P < 0.001) to its previous level of 25 mm Hg. The portal vein pressure has thus proved to be a sensitive gauge of alcohol abstinence or abuse. Lasting, absolute alcohol abstinence is essential in nutritional-toxic liver cirrhosis. PMID- 8428562 TI - [Mixed cryoglobulinemia and vasculitic neuromyopathy]. AB - Over a period of 25 years, a 67-year-old man again and again developed purpura over the lower legs. When hospitalized because of splenomegaly with thrombocytopenia the diagnosis of idiopathic thrombocytopenic purpura was made. Four years later, after an infection of the upper respiratory tract, purpura again developed, together with a symmetrical sensory-motor polyneuropathy with flaccid paraparesis. Biopsy of the N. suralis revealed a florid leukocytoclastic vasculitis. Morphologically there was severe nerve damage with Wallerian degeneration and subtotal nerve fibre loss. Immunological tests demonstrated essential mixed cryoglobulinaemia of IgG/IgM type. Cryoprecipitates must be considered as the cause of the vasculitis, which in turn produced the nerve lesions and purpura. This case underlines the importance of biopsy diagnosis of peripheral nerve and skeletal muscle, because it may demonstrate a treatable cause of a severe acute neuromyopathy. PMID- 8428563 TI - [Public control of contagious animal diseases--effect over changing times]. AB - A survey of the development of public control of animal diseases in Germany is made. The control of contagious diseases on the basis of confederation law has a history of more than hundred years. The knowledge and experience collected during this time are described. PMID- 8428564 TI - [Pharmacotherapy 100 years ago]. AB - In the second part of the 19th century experiments in the sciences, in particular in pharmacology, formed the rational understanding of the present pharmacotherapy. The industry in beginning developed techniques for synthetic pharmaceutical products. A literature study shows that 100 years ago was the time in which the veterinarians get at first the possibility to work with synthetic antiinfective, antipyretic, laxative and anesthetic drugs. PMID- 8428565 TI - [Reading "older" technical literature]. AB - It can be stated, that old technical literature seldom is used for recent publications by modern scientists. But it is necessary to observe old articles not only by historians but even by experts to follow the developments of scientific experience. PMID- 8428566 TI - [The legal regulations of the European Community on animal welfare in the field of agriculture and research]. AB - The European Community has regulated the protection of animals in agriculture and research by means of 9 Directives and one Decision. The agricultural Regulations concern the protection of slaughter animals, the protection of animals during transport and the protection of animals kept for farming purposes as they also cover the protection of laying hens in battery cages and minimum standards for the protection of calves and pigs in intensive farming systems. The protection of animals used for experimental and other scientific purposes is covered by a Directive that approximates the laws, regulations and administrative provisions of the Member States. The legal texts are presented and where necessary commented on. The Directive on experimental animals is compared with the respective passages of the German law on animal protection. PMID- 8428567 TI - [In vitro caffeine test on skeletal muscle samples from swine]. AB - Excitable porcine gracilis muscle samples were subjected to an in vitro caffeine test. Muscle samples were taken from pig carcasses or as biopsy from anaesthetized and halothane tested pigs. The caffeine threshold of the muscle was determined, which was the caffeine concentration of the bath that produced a contracture of the muscle sample. The in vitro caffeine test is used in human medicine to diagnose malignant hyperthermia susceptibility. In this study, the caffeine test was adapted to the particular demands of skeletal muscle samples taken from pig carcasses. The used method is described and the results from 62 pigs are reported. The in vitro caffeine test is a reliable and good reproducible method to diagnose malignant hyperthermia susceptibility in pigs. The caffeine threshold characterizes the resistance of Ca(2+)-ion-storages in the muscle fibers with a numerical value. The caffeine threshold allows a gradual evaluation of the muscle quality and represents a valuable addition to the halothane test. PMID- 8428568 TI - [Sonography of pregnancy in sheep. II. Accuracy of transrectal and transcutaneous pregnancy diagnosis]. AB - Pregnancy was diagnosed by transcutaneous ultrasonography in 327 ewes of 2 flocks. In addition transrectal sonography was performed in 93 of these ewes. Two hundred and thirty six ewes were pregnant and 88 animals were non-pregnant. Criteria to assess the accuracy of transrectal and transcutaneous sonography were overall accuracy (correct diagnoses/all diagnoses), sensitivity (correct diagnoses "pregnant"/all pregnant animals), specificity (correct diagnoses "non pregnant"/all non-pregnant animals), predictive value of positive diagnoses (correct diagnoses "pregnant"/all diagnoses "pregnant"), predictive value of negative diagnoses (correct diagnoses "non-pregnant"/all diagnoses "non pregnant"). Accuracy, sensitivity, specificity, predictive value of positive diagnosis and predictive value of negative diagnosis were 95%, 95%, 91%, 99% and 71% with the transcutaneous sonography resp. 94%, 95%, 93%, 97% and 86% with the transrectal sonography. Accuracy of the transrectal examination was superior (100%; 26/26) to the transcutaneous approach (89%; 39/44) in early pregnancy (Day 22 to 90). Transcutaneous sonography was more appropriate during the last part of pregnancy. The results demonstrate that both methods allow reliable pregnancy diagnosis in sheep under farm conditions. Ultrasonic pregnancy diagnosis is of economical value, if pregnancy rate of herds is below 90% and the diagnostic accuracy reaches at least 95%. PMID- 8428569 TI - [Epidemiologic studies of the course of reproductive events after epidemic late abortion (PRRS/PEARS) in swine breeding herds]. AB - The evaluation of the reproductive performance in nine pig breeding herds that have been affected with PRRS in the beginning of 1991 is carried out by comparing data prior to, during and after the acute phase of PRRS. Piglet losses were increased during four month, performance data, such as percentage of return to oestrus and the interval between farrowings did not show substantial alterations from the pre-epidemic data. The evaluation of reproductive performance of gilts bought in the post-epidemic period did not reveal any depression. Sows which farrowed in the acute epidemic phase of the disease (even those with abortions) returned to their pre-epidemic level of reproductive performance in the following litter. A longlasting immunity due to the persistence of the PRRS virus in the region is discussed as cause for the return of the reproductive performance to the pre-epidemic level. PMID- 8428570 TI - A hundred years of the Deutsche Tierarztliche Wochenschrift. AB - Some reports of veterinary societies were issued in the second half of the 19th century in Germany. Dr. med LYDTIN in Karlsruhe (Baden) tried to unify two local reports to found a new journal with a widespread distribution. The first issue of the "Deutsche Thierarztliche Wochenschrift" was published on 7th January 1893 in Karlsruhe. To increase the area of distribution some years later a professor from the School of Veterinary Medicine Hannover was invited to work as an editor of this journal. After 1897 the DTW was issued in Hannover in close cooperation with the School of Veterinary Medicine. So the DTW is the oldest German veterinary journal which was issued continuously with an unchanged name since 100 years. PMID- 8428571 TI - [Veterinary periodicals 100 years ago--a contribution to the history of the founding of the Deutsche Tierarztliche Wochenschrift]. AB - The role and the meaning of the periodic literature hundred years ago is regarded on the occasion of the jubilee of the "Deutsche tierarztliche Wochenschrift" (1893-1993). A century ago a great restructuring of the veterinarian specialist publications began for the first time due to different scientific and political circumstances. The main attention is directed to two aspects: In the first place the question which periodicals were published in the time of the foundation of the German Empire (1871) until the publication of the DTW (1893). Secondly the question why there was edited one paper after the other and especially why the DTW was founded only five years after the "Berliner tierarztliche Wochenschrift". A brief summary of the prehistory of the DTW concludes the essay. PMID- 8428572 TI - Electron diffraction analysis of structural changes in the photocycle of bacteriorhodopsin. AB - Structural changes are central to the mechanism of light-driven proton transport by bacteriorhodopsin, a seven-helix membrane protein. The main intermediate formed upon light absorption is M, which occurs between the proton release and uptake steps of the photocycle. To investigate the structure of the M intermediate, we have carried out electron diffraction studies with two dimensional crystals of wild-type bacteriorhodopsin and the Asp96-->Gly mutant. The M intermediate was trapped by rapidly freezing the crystals in liquid ethane following illumination with a xenon flash lamp at 5 and 25 degrees C. Here, we present 3.5 A resolution Fourier projection maps of the differences between the M intermediate and the ground state of bacteriorhodopsin. The most prominent structural changes are observed in the vicinity of helices F and G and are localized to the cytoplasmic half of the membrane. PMID- 8428573 TI - Synthetic human beta-globin 5'HS2 constructs function as locus control regions only in multicopy transgene concatamers. AB - Transgenes linked to the beta-globin locus control region (LCR) are transcribed in a copy-dependent manner that is independent of the integration site. It has previously been shown that the LCR 5'HS2 region does not require its NF-E2 dimer binding site for LCR activity. In this paper we analyse synthetic 5'HS2 core constructs containing point mutations in the other factor binding sites 3' of the NF-E2 dimer site. The results show that 5'HS2 core is a partially active LCR that functions in a concatamer of at least two copies but not when present as a single copy in transgenic mice and that no single binding site within 5'HS2 is required for position-independent expression. In addition, the H-BP factor is identical to upstream stimulatory factor (USF) and full enhancement levels by 5'HS2 core in MEL cells require a combination of all the factor binding sites. We suggest that 5'HS2 cores in a concatamer interact with each other to establish an area of open chromatin and that this process may be the basis of LCR function. PMID- 8428574 TI - Novel glucocorticoid receptor complex with DNA element of the hormone-repressed POMC gene. AB - Previous studies defined a DNA element necessary for glucocorticoid repression of the pro-opiomelanocortin (POMC) gene. The glucocorticoid receptor (GR) binds this negative glucocorticoid response element (nGRE) with an in vitro affinity similar to that of GR for positive GREs. However, whereas GR binds GREs as homodimers, a novel GR complex which forms with nGRE appears to contain three GR molecules. Biochemical characterization of this complex as well as equilibrium binding studies suggest that it is formed by sequential binding of a GR homodimer followed by binding of a GR monomer on the opposite side of the double helix. The DNA-binding domain (DBD) of GR is sufficient for differential binding of GRE and nGRE, as bacterially-expressed DBD formed unique nGRE complexes that contain three GR polypeptides. Thus, the POMC nGRE provides the first example of an interaction between GR and DNA in which GR binds otherwise than as a homodimer. Despite its high affinity for GR, the nGRE differs significantly from GREs in that it does not activate transcription in any context. As the nGRE appears insufficient on its own to confer hormone responsiveness, other POMC promoter elements are likely to be required to mediate glucocorticoid repression. PMID- 8428575 TI - Oct-2 facilitates functional preinitiation complex assembly and is continuously required at the promoter for multiple rounds of transcription. AB - Octamer factor 2 (Oct-2, OTF-2, NF-A2) is an 'upstream' promoter factor that binds to the octamer motif (ATGCAAAT) implicated in control of immunoglobulin gene transcription in B-lymphocytes. We have studied the role of Oct-2 in the process of transcription initiation in vitro using both nuclear extracts and purified basal transcription factors. Oct-2 specifically stimulates transcription from octamer-containing promoters in both systems. Thus, Oct-2 is a 'true activator', rather than merely an 'anti-repressor' counteracting the effect of histones. In order-of-addition experiments, Oct-2 is required early, together with TFIID, to allow formation of a preinitiation complex. Oct-2 cannot functionally interact with cloned TATA binding protein (TBP) but rather requires 'coactivators' found in the TFIID fraction. In single-round transcription experiments, early competition for Oct-2 by an octamer oligonucleotide is deleterious, but no effect is seen after assembly of a complete preinitiation complex. However, for multiple rounds of transcription, Oct-2 is continuously required at the promoter; this result argues against a 'hit-and-run' mechanism whereby the activator becomes dispensible after organizing a TFIID-promoter complex. In agreement with our previous studies in vivo, the N-terminal glutamine rich activation domain of Oct-2 is required for full activity in vitro, indicating that this domain directly interacts with basal transcription factors. PMID- 8428576 TI - Dissociation of synthetic Holliday junctions by E. coli RecG protein. AB - The RecG protein of Escherichia coli is needed for normal levels of recombination and for repair of DNA damaged by ultraviolet light, mitomycin C and ionizing radiation. The true extent of its involvement in these processes is masked to a large degree by what appears to be a functional overlap with the products of the three ruv genes. RuvA and RuvB act together to promote branch migration of Holliday junctions, while RuvC catalyses the resolution of these recombination intermediates into viable products by endonuclease cleavage. In this paper, we describe the overproduction and purification of RecG and demonstrate that the overlap extends to the biochemistry. We show that the 76 kDa RecG protein is a DNA-dependent ATPase, like RuvB. Using gel retardation assays we demonstrate that it binds specifically to a synthetic Holliday junction, like RuvA and RuvC. Finally, we show that in the presence of ATP and Mg2+, RecG dissociates these junctions to duplex products, like RuvAB. We suggest that RecG and RuvAB provide alternative activities than can promote branch migration of Holliday junctions in recombination and DNA repair. PMID- 8428577 TI - CDC39, an essential nuclear protein that negatively regulates transcription and differentially affects the constitutive and inducible HIS3 promoters. AB - The yeast HIS3 promoter region contains two functionally distinct TATA elements, TC and TR, that are responsible respectively for initiation from the +1 and +13 sites. Both TC and TR support basal HIS3 transcription and require the TATA binding protein TFIID, but only TR responds to transcriptional activation by GCN4 and GAL4. By selecting for yeast strains that increase transcription by a GCN4 derivative with a defective activation domain, we have isolated a temperature sensitive mutation in CDC39, a previously defined gene implicated in cell-cycle control and the pheromone response. This cdc39-2 mutation causes increased basal transcription of many, but not all genes, as well as increased transcriptional activation by GCN4 and GAL4. Surprisingly, basal HIS3 transcription from the +1 initiation site is strongly increased, while initiation from the +13 site is barely affected. Thus, unlike acidic activator proteins that function through TR, CDC39 preferentially affects transcription mediated by TC. CDC39 is an essential gene that encodes a very large nuclear protein (2108 amino acids) containing two glutamine-rich regions. These observations suggest that CDC39 negatively regulates transcription either by affecting the general RNA polymerase II machinery or by altering chromatin structure. PMID- 8428578 TI - Promoter accessibility within the environment of the MHC is affected in class II deficient combined immunodeficiency. AB - Class II-deficient combined immunodeficiency (CID) is a hereditary disease resulting in abrogation of transcription of the class II genes of the major histocompatibility complex, due to a defect in a trans-acting regulatory factor. Cell lines from certain CID patients lack factor binding at multiple sites in class II promoters in vivo. A mutation in one of the promoter binding proteins could explain this 'bare' phenotype only if these factors bind cooperatively or in a temporal hierarchy. Alternatively, the mutation could affect the configuration of the promoter within the MHC locus. Here, we provide evidence that the factor(s) defective in class II-deficient CID controls the accessibility of class II promoters within the environment of the MHC. The in vivo occupancy of wild type and mutated class II promoter constructs was examined in stable transfectants of normal and CID-derived cell lines. The CID promoter phenotype could not be reproduced in a normal cell line by eliminating binding at any one promoter element, suggesting that these factors bind independently, both spatially and temporally. In contrast, promoter occupancy was partially restored in two CID lines at a randomly integrated wild type promoter, implying that the promoter is inaccessible to factors in its native environment, but accessible when moved to another location in the genome. PMID- 8428579 TI - N-myc suppresses major histocompatibility complex class I gene expression through down-regulation of the p50 subunit of NF-kappa B. AB - In neuroblastoma, N-myc suppresses the expression of major histocompatibility complex (MHC) Class I antigens by reducing the binding of a nuclear factor to the enhancer-A element in the MHC Class I gene promoter. We show here that the p50 subunit of NF-kappa B is part of this complex and that expression of p50 mRNA is suppressed by N-myc. Transfection of a p50 expression vector in neuroblastoma cells that express N-myc at a high level leads to restoration of factor binding to the MHC Class I gene enhancer, restores enhancer activity and leads to re expression of MHC Class I antigens at the cell surface. These data indicate that the p50 subunit of NF-kappa B is involved in the regulation of MHC Class I antigen expression and that N-myc down-regulates MHC Class I gene expression primarily through suppression of p50 expression. PMID- 8428580 TI - The NF-kappa B precursor p105 and the proto-oncogene product Bcl-3 are I kappa B molecules and control nuclear translocation of NF-kappa B. AB - We have examined the interaction of the NF-kappa B precursor p105 with NF-kappa B subunits. Similar to an I kappa B molecule, p105 associates in the cytoplasm with p50 or p65. Through this assembly, p105 efficiently blocks nuclear transfer of either subunit. Moreover, the p105 protein inhibits DNA binding of dimeric NF kappa B subunits in a similar, but not identical, manner to its isolated C terminal domain, which contains an ankyrin-like repeat domain (ARD). The proto oncogene product Bcl-3 also controls nuclear translocation of p50, but not of p65. Hence, p50 can be retained in the cytoplasm via at least three distinct interactions: through direct interactions either with its own precursor, with Bcl 3 or indirectly through I kappa B alpha or -beta when attached to p65. We discuss a function of p105 as a cytoplasmic assembly unit for homo- and heteromeric NF kappa B complexes and of Bcl-3 as an I kappa B with novel subunit specificity. PMID- 8428581 TI - Intermediates in extrachromosomal homologous recombination in Xenopus laevis oocytes: characterization by electron microscopy. AB - Several molecular mechanisms have been proposed to account for nonconservative homologous recombination. This type of recombination is particularly efficient in Xenopus oocytes when appropriate DNA substrates are injected. To distinguish between possible models, we have investigated recombination intermediates from oocytes by direct observation in the electron microscope. Partially recombined DNA was crosslinked with a psoralen derivative after incubation in oocytes to limit the branch migration that might occur during recovery procedures and alter the structures that were initially present. Branched structures, which we interpret as intermediates, represented approximately 10% of the DNA recovered and were readily analyzed. We did not observe any structures with internal loops predicted by invasion mechanisms. The majority of intermediates had one or two single-stranded branches on product-sized molecules, as predicted for incomplete junctions in the resection-annealing mechanism. Detailed length measurements confirmed the expectations of that model. When recovered DNA was not crosslinked, or when annealed junctions were prepared in vitro, we saw branched structures that indicated the occurrence of extensive branch migration. Comparison with the crosslinked sample confirmed the effectiveness of the crosslinking in preserving structures created in the oocytes. Our results strongly support a resection annealing mechanism of recombination in oocytes. PMID- 8428582 TI - Translocation can drive the unfolding of a preprotein domain. AB - Precursor proteins are believed to have secondary and tertiary structure prior to translocation across the Escherichia coli plasma membrane. We now find that preprotein unfolding during translocation can be driven by the translocation event itself. At certain stages, translocation and unfolding can occur without exogenous energy input. To examine this unfolding reaction, we have prepared proOmpA-Dhfr, a fusion protein of the well studied cytosolic enzyme dihydrofolate reductase (Dhfr) connected to the C-terminus of proOmpA, the precursor form of outer membrane protein A. At an intermediate stage of its in vitro translocation, the N-terminal proOmpA domain has crossed the membrane while the folded Dhfr portion, stabilized by its ligands NADPH and methotrexate, has not. When the ligands are removed from this intermediate, translocation occurs by a two-step process. First, 20-30 amino acid residues of the fusion protein translocate concomitant with unfolding of the Dhfr domain. This reaction requires neither ATP, delta mu H+ nor the SecA subunit of translocase. Strikingly, this translocation accelerates the net unfolding of the Dhfr domain. In a second step, SecA and ATP hydrolysis drive the rapid completion of translocation. Thus energy derived from translocation can drive the unfolding of a substantial protein domain. PMID- 8428583 TI - The SecA and SecY subunits of translocase are the nearest neighbors of a translocating preprotein, shielding it from phospholipids. AB - To study the environment of a preprotein as it crosses the plasma membrane of Escherichia coli, unique cysteinyl residues were introduced into proOmpA and the genes for these mutant preproteins were fused to the gene of dihydrofolate reductase (Dhfr). A photoactivable, radiolabeled and reducible cross-linker was then attached to the unique cysteinyl residue of each purified protein. Partially translocated polypeptides were generated and arrested in their membrane transit by the folded structure of the dihydrofolate reductase domain. After photolysis to label their nearest neighbors and reduction of the disulfide bond between proOmpA-Dhfr and the cross-linker, radiolabeled cross-linker was selectively recovered with the SecA and SecY subunits of preprotein translocase. Strikingly, neither the SecE nor Band 1 subunits were cross-linked to any of the constructs and the membrane phospholipids were almost entirely shielded from cross-linking. The fact that SecY and SecA are the only membrane proteins cross-linked to the translocating chains suggests that they may form an entirely proteinaceous pathway through which secreted proteins pass during membrane transit. PMID- 8428584 TI - SecD is involved in the release of translocated secretory proteins from the cytoplasmic membrane of Escherichia coli. AB - The SecD protein is one of the components that has been suggested from genetic studies to be involved in the protein secretion across the cytoplasmic membrane of Escherichia coli. We examined the effect of anti-SecD IgG on protein secretion using spheroplasts. Inhibition of the secretion of OmpA and maltose-binding protein (MBP) by this IgG was observed with concomitant accumulation of their precursor and mature forms in spheroplasts. This effect was specific to anti-SecD IgG. Anti-SecE and anti-SecY IgGs, of which the epitopes are located at the periplasmic domains of SecE and SecY, respectively, did not interfere with the secretion. Time-course experiments investigating the processing of proMBP and the release of MBP from spheroplasts revealed that anti-SecD IgG interfered with the release of the translocated mature MBP. The mature form of MBP thus accumulated was sensitive to trypsin, which was externally added to spheroplasts, whereas MBP released into the medium was resistant to trypsin as the native MBP is. The precursor form of MBP accumulated in spheroplasts was also trypsin resistant. We conclude that SecD is directly involved in protein secretion and important for the release of proteins that have been translocated across the cytoplasmic membrane. PMID- 8428585 TI - Stable periplasmic secretion intermediate in the general secretory pathway of Escherichia coli. AB - The secretion of the Klebsiella oxytoca cell surface lipoprotein pullulanase involves translocation across the cytoplasmic and outer membranes of the Gram negative bacterial cell envelope. A variant of pullulanase was created by fusing the signal peptide-encoding 5' region of the Escherichia coli gene for periplasmic MalE protein to the 3' end of the pulA gene encoding almost the entire mature part of pullulanase. When produced in E. coli carrying the malE pulA gene fusion on a high copy number plasmid and the complete set of genes specifically required for pullulanase secretion on a second plasmid, the hybrid protein differed from wild-type pullulanase as follows: (i) it was not fatty acylated; (ii) it was apparently processed by LepB signal peptidase rather than by LspA lipoprotein signal peptidase; (iii) it was released into the periplasm and was only slowly transported across the outer membrane, and (iv) it was released directly into the medium rather than via the usual surface-anchored intermediate. The hybrid protein was secreted more rapidly when malE-pulA was expressed from a low copy number plasmid. The two steps in the secretion pathway could be totally uncoupled by expressing first the malE-pulA gene fusion and then the cognate secretion genes. These results show that fatty-acylation of wild-type PulA is not essential for secretion but may improve its efficiency when large amounts of the protein are produced, that the two steps in secretion can occur quite independently and that the periplasmic intermediate can persist for long periods under certain circumstances. PMID- 8428586 TI - Yeast Wbp1p and Swp1p form a protein complex essential for oligosaccharyl transferase activity. AB - Asparagine-linked N-glycosylation is an essential protein modification occurring in all eukaryotic cells. The central step is the co-translational transfer of the core oligosaccharide assembled on the lipid carrier dolichol phosphate to selected Asn-X-Ser/Thr residues of nascent polypeptide chains in the endoplasmic reticulum. This reaction is catalyzed by the enzyme N-oligosaccharyl transferase. In yeast, Wbp1p is an essential component of this enzyme. Using a high copy number suppression approach, the SWP1 gene was isolated as an allele specific suppressor of a wbp1 mutation. Swp1p is a 30 kDa type I transmembrane protein and essential for cell viability. Similar to Wbp1p, depletion of Swp1p results in reduced N-oligosaccharyl transferase activity in vivo and in vitro. Wbp1p and Swp1p can be chemically cross-linked, suggesting that both proteins are essential constituents of the N-oligosaccharyl transferase complex. PMID- 8428587 TI - Lysosomal aspartylglucosaminidase is processed to the active subunit complex in the endoplasmic reticulum. AB - Aspartylglucosaminidase (AGA) is a lysosomal enzyme, the deficiency of which leads to a human storage disease, aspartylglucosaminuria (AGU). Although numerous mutations have been identified in AGU patients, elucidation of the molecular pathogenesis of the disease has been hampered by the missing information on the cellular events resulting in the maturation and activation of the enzyme. Here we used the expression of in vitro mutagenized constructs of the AGA cDNA to define three specific proteolytic trimming steps resulting in mature AGA. Removal of the signal peptide is immediately followed by proteolytic cleavage of the precursor into two subunits and results in biologically active enzyme already in the endoplasmic reticulum. This early activation has not previously been described for lysosomal enzymes. The subsequent lysosomal trimming does not influence the enzymatic activity of AGA. It consists only of a single proteolytic cleavage which removes 10 amino acids from the C-terminal end of the larger subunit, in contrast to the multistep lysosomal processing observed in several other hydrolases. PMID- 8428588 TI - Millisecond studies of secretion in single rat pituitary cells stimulated by flash photolysis of caged Ca2+. AB - To study the final steps in the secretory pathway of rat pituitary melanotrophs, we have monitored changes in cell surface area due to exocytosis after flash photolysis of caged Ca2+. A step increase in cytosolic [Ca2+] to 45-125 microM triggers three phases of exocytic secretion. A small cohort of a few hundred vesicles is exocytosed in 40 ms in a secretory burst with a peak rate of 17,000 vesicles/s. Next, 2700 more vesicles are released in a slower phase that is complete within 400-1000 ms. Finally, vesicles continue to be released slowly (500 vesicles/s) for > 8s. The approach described provides a way to identify and monitor the final steps in the secretory pathway at millisecond resolution. That a small portion of secretory vesicles can be released much faster than all others suggests that these vesicles are functionally equivalent to those at the presynaptic active zone of a neuron. Their release would be fast enough to be temporally correlated with single action potentials. PMID- 8428589 TI - CD45 specifically modulates binding of Lck to a phosphopeptide encompassing the negative regulatory tyrosine of Lck. AB - CD45 is a tyrosine phosphatase expressed in all hematopoietic cells which is important for signal transduction through the T cell antigen receptor (TCR). Studies using CD45-deficient cells have revealed that Lck, a tyrosine kinase thought to be essential for TCR signaling, is hyperphosphorylated on Y505 in the absence of CD45. This site of tyrosine phosphorylation negatively regulates the function of the Src family of kinases. Here we provide evidence that CD45 can modulate the binding of the Lck to an 11 amino acid tyrosine phosphorylated peptide containing the carboxy-terminus of Lck (lckP). Significantly, CD45 did not influence the binding of Fyn, PLC gamma 1, GAP and Vav to the same phosphopeptide. Lck protein which bound the peptide was dephosphorylated on Y505 and consisted of only 5-10% of the total cellular Lck. Interestingly, there was a marked increase in binding 15-30 min after CD4 or TCR cross-linking. Taken together, our data suggest that CD45 specifically modulates the conformation of Lck in a manner consistent with the intramolecular model of regulation of Src like kinases. PMID- 8428590 TI - Variation of half-site organization and DNA looping by AraC protein. AB - The dimeric AraC protein of Escherichia coli binds specifically to DNA sequences upstream of promoters whose transcription is regulated by arabinose. Here we show with affinity measurements, DNase footprinting, dimethyl sulfate premethylation interference and dimethyl sulfate footprinting studies that AraC protein can recognize paired half-sites in direct repeat orientation or inverted repeat orientation. A similar high degree of flexibility was also seen in the ability of the protein in the absence of arabinose to bind tightly and specifically when the separation of its half-sites was increased by 10 or 21 bp. In the presence of arabinose the protein could specifically contact both half-sites of a +10 bp spacing construct but could not contact both in a +21 bp construct. Reduced extensibility of AraC protein in the presence of arabinose provides a simple mechanism for the protein's shift from a non-inducing, DNA looping state to an inducing, non-looping state that contacts two adjacent half-sites at the arapBAD promoter. PMID- 8428591 TI - Clonal deletion of specific thymocytes by an immunoglobulin idiotype. AB - We have investigated whether immunoglobulin can induce clonal deletion of thymocytes by employing two strains of transgenic mice. One strain is transgenic for an alpha/beta T cell receptor (TCR) which recognizes a processed idiotypic peptide of the lambda 2(315) light chain variable region, bound to the I-Ed class II major histocompatibility complex molecule. The other mouse strain is transgenic for the lambda 2(315) gene. Double transgenic offspring from a TCR transgenic female mated with a lambda 2(315) transgenic male exhibit a pronounced clonal deletion of CD4+CD8+ thymocytes. Analysis of neonates from the reciprocal (lambda 2(315)-transgenic female x TCR-transgenic male) cross suggests that the deletion in double transgenic offspring most likely is caused by lambda 2(315) produced within the thymus rather than by maternally derived IgG, lambda 2(315). Nevertheless, IgG, lambda 2(315) can cause deletion of CD4+CD8+ thymocytes when injected in large amounts intraperitoneally into either adult or neonatal TCR transgenic mice. Deletion is evident 48 and 72 h after injection, but by day 7 the thymus has already regained its normal appearance. A serum concentration of several hundred microgram/ml is required for deletion to be observed. Therefore, the heterogeneous idiotypes of serum Ig are probably each of too low concentration to cause thymocyte deletion in normal animals. PMID- 8428592 TI - Characterization of the endogenous deoxyribonuclease involved in nuclear DNA degradation during apoptosis (programmed cell death). AB - Cell death by apoptosis occurs in a wide range of physiological events including repertoire selection of lymphocytes and during immune responses in vivo. A hallmark of apoptosis is the internucleosomal DNA degradation for which a Ca2+,Mg(2+)-dependent endonuclease has been postulated. This nuclease activity was extracted from both rat thymocyte and lymph node cell nuclei. When incubated with nuclei harbouring only limited amounts of endogenous nuclease activity, the ladder pattern of DNA fragments characteristic of apoptosis was induced. This extractable nucleolytic activity was immunoprecipitated with antibodies specific for rat deoxyribonuclease I (DNase I) and was inhibited by actin in complex with gelsolin segment 1, strongly pointing to the presence of a DNase I-type enzyme in the nuclear extracts. COS cells transiently transfected with the cDNA of rat parotid DNase I expressed the enzyme, and their nuclei were able to degrade their DNA into oligosome-sized fragments. PCR analysis of mRNA isolated from thymus, lymph node cells and kidney yielded a product identical in size to that from rat parotid DNase I. Immunohistochemical staining with antibodies to rat DNase I confirmed the presence of DNase I antigen in thymocytes and lymph node cells. The tissue distribution of DNase I is thus extended to tissues with no digestive function and to cells which are known to be susceptible to apoptosis. We propose that during apoptosis, an endonuclease indistinguishable from DNase I gains access to the nucleus due to the breakdown of the ER and the nuclear membrane. PMID- 8428593 TI - Origin recognition specificity in pT181 plasmids is determined by a functionally asymmetric palindromic DNA element. AB - The leading strand replication origin of pT181 plasmids consists of two adjacent inverted repeat elements (IR-II and IR-III), which are involved in origin recognition by the initiator (Rep) protein. The conserved core element, IR-II, which contains the initiation nick site, is induced by Rep to form a cruciform structure, probably the primary substrate for the initiation of rolling circle replication. The divergent repeat, IR-III, constitutes the determinant of origin recognition specificity. We show here that the distal arm of IR-III is not required for sequence-specific recognition, whereas the proximal arm and central region are required. Since the initiator is dimeric, we presume that it binds symmetrically to IR-III. A unique type of DNA-protein interaction is proposed, in which the lack of sequence requirement for the distal arm is a consequence of binding to the adjacent IR-II, which thereby polarizes the stringency of binding to the two arms of IR-III. In addition, genetic evidence indicates that both the spacing and the phasing of IR-II to IR-III are crucial for function and that the central segment of IR-III may serve to position the two flanking half-sites for optimal interaction of Rep with IR-III. PMID- 8428594 TI - Phosphorylation and activation of human cdc25-C by cdc2--cyclin B and its involvement in the self-amplification of MPF at mitosis. AB - We have investigated the mechanisms responsible for the sudden activation of the cdc2-cyclin B protein kinase before mitosis. It has been found previously that cdc25 is the tyrosine phosphatase responsible for dephosphorylating and activating cdc2-cyclin B. In Xenopus eggs and early embryos a cdc25 homologue undergoes periodic phosphorylation and activation. Here we show that the catalytic activity of human cdc25-C phosphatase is also activated directly by phosphorylation in mitotic cells. Phosphorylation of cdc25-C in mitotic HeLa extracts or by cdc2-cyclin B increases its catalytic activity. cdc25-C is not a substrate of the cyclin A-associated kinases. cdc25-C is able to activate cdc2 cyclin B1 in Xenopus egg extracts and to induce Xenopus oocyte maturation, but only after stable thiophosphorylation. This demonstrates that phosphorylation of cdc25-C is required for the activation of cdc2-cyclin B and entry into M-phase. Together, these studies offer a plausible explanation for the rapid activation of cdc2-cyclin B at the onset of mitosis and the self-amplification of MPF observed in vivo. PMID- 8428595 TI - Synergistic action of Drosophila cyclins A and B during the G2-M transition. AB - A variety of different cyclin proteins have been identified in higher eukaryotes. In the case of cyclin B, functional analyses have clearly demonstrated an important role in the control of entry into mitosis. The function of cyclin A is more complex. It appears to function in the control of both S- and M-phase. The results of our genetic analyses in Drosophila demonstrate that cyclin A has a mitotic function and that it acts synergistically with cyclin B during the G2-M transition. In double mutant embryos that express neither cyclin A nor cyclin B zygotically, cell cycle progression is blocked just before the exhaustion of the maternally contributed cyclin A and B stores. BrdU-labeling experiments indicate that cell cycle progression is blocked in G2 before entry into the fifteenth round of mitosis. Expression of either cyclin A or B from heat-inducible transgenes is sufficient to overcome this cell cycle block. This block is also not observed in single mutant embryos deficient for either cyclin A or B. In cyclin B deficient embryos, cell cycle progression continues after the apparent exhaustion of the maternal contribution, suggesting that cyclin B might not be essential for mitosis. However, mitotic spindles are clearly abnormal and progression through mitosis is delayed in these cyclin B deficient embryos. PMID- 8428596 TI - Human Wee1 kinase inhibits cell division by phosphorylating p34cdc2 exclusively on Tyr15. AB - In fission yeast, the M-phase inducing kinase, a complex of p34cdc2 and cyclin B, is maintained in an inhibited state during interphase due to the phosphorylation of Cdc2 at Tyr15. This phosphorylation is believed to be carried out primarily by the Wee1 kinase. In human cells the negative regulation of p34cdc2/cyclin B is more complex, in that Cdc2 is phosphorylated at two inhibitory sites, Thr14 and Tyr15. The identities of the kinases that phosphorylate these sites are unknown. Since fission yeast Wee1 kinase behaves as a dual-specificity kinase in vitro, a popular hypothesis is that a human Wee1 homolog might phosphorylate p34cdc2 at both sites. We report here that a human gene, identified as a possible Wee1 homologue, blocks cell division when overexpressed in HeLa cells. This demonstrates functional conservation of the Wee1 mitotic inhibitor. Contrary to the dual-specificity kinase hypothesis, purified human Wee1 phosphorylates p34cdc2 exclusively on Tyr15 in vitro; no Thr14 phosphorylation was detected. Human and fission yeast Wee1 also specifically phosphorylate synthetic peptides at sites equivalent to Tyr15. Mutation of a critical lysine codon (Lys114) believed to be essential for kinase activity abolished both the in vivo mitotic inhibitor function and in vitro kinase activities of human Wee1. These results conclusively prove that Wee1 kinases inhibit mitosis by directly phosphorylating p34cdc2 on Tyr15, and strongly indicate that human cells have independent kinase pathways directing the two inhibitor phosphorylations of p34cdc2. PMID- 8428597 TI - The inactive form of recA protein: the 'compact' structure. AB - When recA protein is enzymatically inactive in vitro, it adopts a more compact helical polymer form than that of the active protein polymerized onto DNA in the presence of ATP. Here we describe some aspects of this structure. By cryo electron microscopy, a pitch of 76 A is found for both the self-polymer and the inactive complex with ssDNA. A smaller pitch of 64 A is observed in conventional electron micrographs. The contour length of complexes with ssDNA was used to estimate the binding stoichiometry in the compact complex, 6 +/- 1 nt/recA. In addition, the compact structure was observed in vivo in Escherichia coli: inclusion bodies produced upon induction of recA expression in an overproducing strain have a fibrous morphology with the structural parameters of the compact polymer. PMID- 8428598 TI - Motor effects of indomethacin, morphine or vagal nerve stimulation on the feline small intestine in vivo. AB - Some factors known to affect jejunal motility (recorded as volume changes of an intraluminal balloon) were investigated in anaesthetized cats (ether-chloralose) pretreated with guanethidine and atropine. Indomethacin, morphine (both compounds administered systemically) or vagal nerve stimulation elicited jejunal excitatory motor responses. The effect of indomethacin seemed to be independent of cyclooxygenase inhibition and probably did not involve opioid receptors. It is suggested that the spasmogenic stimuli caused jejunal hypermotility by inhibiting tonically active, inhibitory motor neurons that are intrinsic to the gut. Furthermore, when the jenunal tone had been raised by indomethacin or morphine spontaneous relaxations were observed, and these could be mimicked by vagal stimulation. Hexamethonium antagonized these relaxations but did not attenuate the drug-induced jejunal hypermotility. PMID- 8428599 TI - 5-Hydroxytryptamine stimulates human isolated atrium but not ventricle. AB - Although 5-hydroxytryptamine (5-HT) elicits positive inotropic effects in the human isolated atrium via 5-HT4 receptors, no data are available about its effects in the ventricular myocardium. We investigated the inotropic effects of 5 HT in human healthy ventricular trabeculae and compared these effects with those in right atrial trabeculae. Baseline force of contraction as well as the response to noradrenaline, used to check inotropic responsiveness of the tissue, was significantly higher in ventricular (391 +/- 10 and 719 +/- 126 mg, respectively) than in atrial tissue (189 +/- 5 and 383 +/- 79 mg, respectively). However, 5-HT increased the force of contraction up to 309 +/- 82 mg at 10(-4) M in atrial trabeculae, but failed to affect the force of contraction in ventricular tissue. We conclude that, in contrast to atrial tissue, 5-HT is ineffective as a positive inotropic agent in human ventricular trabeculae. This finding obviously rules out the development of 5-HT4 receptor agonists for the treatment of heart failure, but suggests the absence of ventricular side-effects of 5-HT4 receptor (ant)agonists should these agents be used in the treatment of gastrointestinal disorders. PMID- 8428600 TI - Lysergic acid diethylamide is a partial agonist at 5-HT2 receptors in ovine uterine artery of late pregnancy. AB - d-Lysergic acid diethylamide (LSD) produced dose-dependent contractions (EC50, 17.9 +/- 2.1 nM) on isolated ovine uterine artery of late pregnancy, which were competitively antagonized by ketanserin. The maximal contraction to LSD was 51% of the 5-HT response. LSD competitively antagonized (pA2 9.21) contractions produced to 5-hydroxytryptamine (5-HT). The results indicate that LSD is a partial agonist at 5-HT2 receptors in ovine uterine artery. PMID- 8428601 TI - Effect of NMDA receptor antagonists on rapid tolerance to ethanol. AB - Hypothermia and motor impairment (tilt-plane test) were used to assess whether N methyl-D-aspartate (NMDA) receptors play a role in the development of rapid tolerance to ethanol, i.e., tolerance to a second dose of ethanol given 24 h after the effect of the first dose of ethanol had disappeared. Results showed that (+)-MK-801 and ketamine blocked the development of rapid tolerance to ethanol on both tests. Since these drugs did not modify blood or brain alcohol levels in any of the groups, the blockade of ethanol rapid tolerance cannot be attributed to changes in pharmacokinetics of ethanol. These data suggest that the role of NMDA receptors in ethanol tolerance may be similar to their role in memory and learning, involving a facilitation of transmission in certain synapses. PMID- 8428602 TI - Inhibition of tubular lithium reabsorption by amiloride in the conscious sodium restricted rat. AB - We examined the effects of amiloride administration on the renal lithium clearance (CLi) in three series of conscious, unrestrained rats maintained on either a low (5 mmol/kg) or normal (200 mmol/kg) sodium diet. In series 1, six doses of amiloride (0.5-16 mg/kg) were administered s.c., and the renal electrolyte excretion was assessed over a 3-h clearance period. In series 2, the time profile of changes in renal electrolyte excretion following 4 mg/kg of amiloride s.c. was examined, and in series 3, the effect of amiloride infusion i.v. (1 mg/kg followed by 2 mg/kg per h) on renal function was investigated. In all series CLi was lower in sodium-restricted rats than in controls. Amiloride administered s.c. to sodium-restricted rats did not increase the 3-h CLi to the levels found in control rats. When amiloride was administered s.c. or i.v. and urine collected in 30-minute periods, CLi in rats fed a sodium-deficient diet increased to control levels. We conclude that amiloride-induced losses of sodium and water in CLi studies may lead to an erroneous interpretation of data. However, distal tubular lithium reabsorption may be recognized, if present, by the administration of amiloride i.v. or s.c. during collections of urine in short time intervals. PMID- 8428603 TI - Effect of BRL 38227 on the adrenergic regulation of the kidney vasculature of the rat. AB - Sodium pentobarbitone anaesthetised Wistar rats were prepared for the measurement of renal blood flow, close renal-arterial infusion of drugs and electrical stimulation of the renal nerves. Administration of intra-renal arterial bolus doses of noradrenaline, phenylephrine, methoxamine and renal nerve stimulation caused transient dose- or frequency-related reductions in renal blood flow, ranging from 20% to 85%, with all slopes being significant (P < 0.05-0.01). Intra arterial infusion of BRL38227 for 30 min at 0.5 micrograms/kg per min had no effect on blood pressure but at 0.75 and 1.0 micrograms/kg per min there were reductions of 9 mm Hg (P < 0.05) and 19 mm Hg (P < 0.01), respectively. The slopes of the dose-response curves for noradrenaline, phenylephrine and methoxamine and frequency-response curves for renal nerve stimulation generated at each dose level of BRL38227 could not be distinguished statistically. The findings of this investigation suggest that the adrenergic control of renal haemodynamics would occur normally in the presence of hypotensive doses of BRL 38227. PMID- 8428604 TI - Effects of thapsigargin and ryanodine on vascular contractility: cross-talk between sarcoplasmic reticulum and plasmalemma. AB - Thapsigargin and ryanodine are proposed to interfere with Ca2+ storage in sarcoplasmic reticulum by different mechanisms. Thapsigargin inhibits Ca2+ transport into and ryanodine enhances Ca2+ out of the sarcoplasmic reticulum. Contractility studies were performed in the rat aorta and dog mesenteric artery. Ryanodine was found to reduce phenylephrine-induced (10 microM) contraction in Ca(2+)-free medium of rat aorta and dog mesenteric artery in a concentration dependent manner. Each agent alone caused a slow contraction in the rat aorta. In this tissue, the tension caused by ryanodine (30 microM) but not that by thapsigargin (1 microM) was found to be dependent on the status of the sarcoplasmic reticulum: prior stimulation with K+ (60 mM) enhanced the rate of development of ryanodine-induced tension compared with when the sarcoplasmic reticulum was previously depleted with phenylephrine stimulation in Ca(2+)-free medium. Sodium nitroprusside (1 microM) or isoproterenol (1 microM) fully antagonized the contraction induced by ryanodine or phenylephrine. However, thapsigargin-induced contraction was antagonized fully by sodium nitroprusside and only partially by isoproterenol. This result suggests that cAMP elevation by isoproterenol required a functioning sarcoplasmic reticulum Ca2+ pump for its relaxant effect while cGMP elevation by sodium nitroprusside did not. These findings are consistent with the view that ryanodine promotes Ca2+ release from the sarcoplasmic reticulum and that thapsigargin inhibits the ability of cAMP to stimulate Ca2+ uptake into the store by blocking its Ca2+ pump. In the dog mesenteric artery, when the phenylephrine-sensitive Ca2+ pool was emptied and thapsigargin was added to block Ca2+ uptake into the store, restoration of Ca2+ in the Ca(2+)-free medium caused a transient contraction (absent in controls). This contraction was replaced by a significantly larger amplitude and more sustained contraction in low Na+ medium indicating the involvement of the Na+/Ca2+ exchanger in the homeostasis of cytosolic [Ca2+]. In the presence of nifedipine (2 microM), repletion of the phenylephrine-sensitive store was inhibited. It is possible that refilling occurs in part through L-type Ca2+ channels. PMID- 8428605 TI - Two components in the adrenal nicotinic secretory response revealed by cobalt ramps. AB - Prolonged stimulation with nicotine (50 microM) enhanced the secretion of catecholamines from perfused cat adrenal glands. The profile of secretion consisted of a quick activation phase to a peak of 7.68 micrograms/min followed by a second inactivation phase which exhibited a t1/2 of 3.75 min. Sustained stimulation with a solution enriched in K+ (59 mM) also evoked a transient secretory response, with a peak release of 8.62 micrograms/2 min and a t1/2 for inactivation of 4.8 min. Co2+ (10 mM) blocked the nicotinic response by 58% and the K(+)-evoked secretory response by over 96%. In the presence of Co2+ (5 mM), continuous perfusion with nicotine produced a transient but large initial secretory response; the gradual decrease of the extracellular Co2+ concentration, [Co2+]0, as a continuous ramp allowed the development of a second component of secretion which inactivated later on. When the glands were continuously stimulated with 59 mM K+ in the presence of Co2+, the first component of secretion was missing; the second component appeared as [Co2+]0 decreases as a ramp. In similar experiments performed in low-Na+ solution (10 mM Na+), only the first secretion component evoked by nicotine was observed. This finding suggests that the second component of secretion depends on Na+ entry through the nicotinic receptor, on the ensuing cell depolarization and on Ca2+ entry through voltage dependent Ca2+ channels.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428606 TI - Swiss Academy of Medical Sciences and Swiss Academy of Sciences. Ethical principles and guidelines for experiments on animals. PMID- 8428607 TI - Transgenic animal models of sickle cell disease. AB - An animal model which allows study of chronic processes occurring in sickle cell disease has finally been realized with the development of several lines of transgenic mice which express high levels of beta s or beta s-variants in their red cells. The red cells of all mouse lines exhibit characteristic sickle shapes on deoxygenation and most lines have enlarged spleens and mildly elevated reticulocytes in adult mice; both of these properties are suggestive of enhanced red cell destruction and erythropoiesis. However, all lines examined to date have normal hemoglobin levels in adult mice. In one mouse line under ambient conditions, retinopathy and abnormal renal function have been observed and in the same line under hypoxic conditions, anemia, irreversibly sickled cell formation, and urine concentrating defect have been observed. The current mouse lines will allow some aspects of sickle cell disease to be studied, but significant improvements can still be made in the transgenic mouse model. PMID- 8428608 TI - Transgenic and mutant animal models to study mechanisms of protection of red cell genetic defects against malaria. AB - Malaria, caused by members of the genus Plasmodia, is still the most prevalent parasitic disease in the world. In an attempt to understand genetic factors conferring resistance to malaria, mouse models of thalassemia, sickle trait, and ankyrin and spectrin deficiency were studied during infection with species of malaria infectious to rodents. Although growth of P. falciparum is not inhibited in thalassemic erythrocytes in culture, mice carrying a beta-thalassemia mutation were protected from Plasmodium chabaudi adami, supporting epidemiologic findings. Transgenic mice expressing beta s hemoglobin were also significantly protected from two species of rodent malaria. Importantly, a significant role for the spleen in protection in the beta s transgenic mice was found. Finally, mice deficient in spectrin and ankyrin were studied with respect to their ability to support the growth of malaria. It was found that spectrin deficient mice were almost completely refractory to P. chabaudi adami and P. berghei. These models will allow further study of host factors in resistance to malaria. PMID- 8428609 TI - Developments in sickle cell anemia research. Introduction. PMID- 8428610 TI - Triphasic vascular effects of thiol compounds and their oxidized forms on dog coronary arteries. AB - The vascular effects of 2-mercaptoethanol, cysteamine, L-cysteine, glutathione (GSH), cystamine and oxidized GSH (GSSG) on the isometric tension of isolated dog coronary arterial strips were examined, and these effects were compared with the triphasic response induced by dithiothreitol (DTT); a rapid and weak contraction (phase A), an intervening slow relaxation (phase B) and a slowly-developing strong contraction (phase C) which we previously reported. The responses of the arteries induced by 2-mercaptoethanol, cysteamine and L-cysteine consisted of phases A, B and C. The order of contractile potency (ED50 of phase C) was DTT approximately L-cysteine > 2-mercaptoethanol approximately cysteamine, while the order of relaxant potency (ED50 of phase B) was DTT > cysteamine approximately 2 mercaptoethanol. GSSG and cystamine mainly produced relaxation, which corresponded to phase B. The phase C contraction was specific to the reduced forms of thiols, except for GSH, which produced only relaxation. The participation of endothelial cells was not essential for the contracting or relaxing effects of the thiol compounds. The phase C contraction was depressed by W-7, a calmodulin antagonist, while phase A was not. Therefore calmodulin dependent protein kinases may participate in phase C, not in phase A. PMID- 8428612 TI - Heat shock proteins in three related Drosophila species belonging to the obscura group. AB - The effect of heat shock on protein synthesis in three related Drosophila species belonging to the obscura group was analyzed on SDS-acrylamide gels. Four major heat shock proteins (hsps) were found in these species, in which synthesis reaches a maximum at 34 degrees C. Although the higher molecular weight proteins are conserved, differences in size were found for the small hsps in these species. By means of in situ hybridization using D. melanogaster probes for the small hsp genes, it was inferred that the small hsp genes of the obscura group species are clustered at the 27A locus in all three species. PMID- 8428611 TI - Sickle cell vasoocclusion: many issues and some answers. AB - The pathophysiology of sickle (SS) cell vasoocclusion is derived from the presence of hemoglobin S (HbS) which forms polymeric fibers in the deoxygenated state. Nevertheless, phenotypic expression of sickle cell disease (i.e., clinical severity) shows marked individual variations and is influenced by genetic modifiers such as epistatic effects of linked and unlinked genes. Furthermore, the polymerization of HbS is central but not the only event, and is more likely a consequence of disruptions of the steady state of flow. The available evidence indicates that the vasoocclusive crisis is a microcirculatory event in which multiple factors could be involved. We present a model of vasoocclusion as a two step process in which adhesion of deformable cells occurs first, followed by obstruction induced by less deformable SS cells. This review discusses, in addition, rheologic and microcirculatory behavior of SS erythrocytes and the interacting role of vascular factors, red cell heterogeneity, deoxygenation rates, and red cell-endothelial interactions in the pathophysiology of SS cell vasoocclusion. PMID- 8428613 TI - Dopamine regulation of testicular activity in intact and hypophysectomized frogs, Rana esculenta. AB - In intact frogs, both GnRHA and L-dopa were able to increase testicular and plasma androgen levels and to induce spermiation. The dopamine antagonist pimozide inhibited both the effects of L-dopa but not those of GnRHa. Hypophysectomy reduced androgen levels, but spermiation was still induced by both GnRHa and L-dopa, suggesting that these agents can directly influence the testis through a route not involving the pars distalis. Again, pimozide antagonised spermiation induced by L-dopa but not that induced by GnRHa. PMID- 8428614 TI - Effects of ovarian steroids on superoxide dismutase activity in the rat brain. AB - The levels of manganese superoxide dismutase (MnSOD) and copper-zinc superoxide dismutase (CuZn-SOD) were determined in appropriate subcellular fractions prepared from whole brain homogenates of cycling and long-term (3 week) ovariectomized (OVX) Wistar rats, and were compared to the levels found in corresponding samples prepared from OVX rats treated with progesterone (P) or estradiol 17B-benzoate (EB). The activity of both SODs was steady during the estrous cycle, except at proestrus, when MnSOD activity was elevated significantly. Bilateral ovariectomy resulted three weeks later in an increase of the MnSOD activity even higher than that recorded at proestrus. High post castration MnSOD activity was lowered profoundly by exogenous P (2 mg) or EB (0.5 micrograms), given s.c. to OVX animals 2 h or 24 h before sacrifice. Neither removal of the ovaries nor the hormone treatments affected the activity of CuZnSOD. These results suggest suppressive effects of ovarian steroids on MnSOD activity in the rat brain. PMID- 8428615 TI - Two phorbol ester receptor affinities in partially transformed human urothelial cells and decrease of receptor binding in desensitized cells. AB - The presence of specific binding sites for phorbol esters was studied in a transformed but non-tumorigenic human urothelial cell line HCV-29 by assay of specific binding of 3H-phorbol-12,13-dibutyrate (3H-PDBu) to intact living cells. 3H-PDBu bound specifically to HCV-29 cells in a saturable and competitive manner. Scatchard plot analysis of specific binding yielded a curved plot consistent with two binding sites with Kd of 11 nM and 102 nM, respectively. At saturation the corresponding PDBu binding capacities (Bmax) were 8.8 pmol/10(6) cells (5.2 x 10(6) molecules bound per cell) and 2.8 pmol/10(6) cells (1.7 x 10(6) molecules bound per cell). 3H-PDBu binding was displaced by biologically active phorbol ester tumor promoters such as 12-O-tetradecanoylphorbol-13-acetate (TPA) and mezerein, but not by tumor promoters such as L-tryptophan, anthranilic acid and sodium saccharin. In cells desensitized by pretreatment with 1 microgram/ml (2 microM) TPA or PDBu for 24 h the level of binding was reduced to 28% of the level in non-exposed cells. The ability of desensitized cells to bind 3H-PDBu was gradually restored within 5-6 days. At the same time the cells became sensitive to the morphological alteration induced by PDBu. This suggests that desensitization of HCV-29 cells is due to a decreased receptor-ligand binding capacity probably associated with down regulation of the phorbol ester receptors. PMID- 8428616 TI - Erythrophagocytosis by cultured skin fibroblasts from patients with hereditary metabolic disorders. AB - Phagocytosis of native allogenic red blood cells was observed in cultures of skin fibroblasts obtained from patients with neuronal ceroid-lipofuscinosis, Niemann Pick disease type C and morbus Fabry. Occasional phagocytizing cells were observed in 9 other syndromes. Cells from three normal donors did not phagocytize. PMID- 8428617 TI - Naturally occurring deuterium is essential for the normal growth rate of cells. AB - The role of naturally occurring D in living organisms has been examined by using deuterium-depleted water (30-40 ppm D) instead of water containing the natural abundance of D (150 ppm). The deuterium-depleted water significantly decreased the growth rate of the L929 fibroblast cell line, and also inhibited the tumor growth in xenotransplanted mice. Eighty days after transplantation in 10 (59%) out of 17 tumorous mice the tumor, after having grown, regressed and then disappeared. We suggest that the naturally occurring D has a central role in signal transduction involved in cell cycle regulation. PMID- 8428618 TI - Crystallization and preliminary crystallographic analysis of trypanothione reductase from Trypanosoma cruzi, the causative agent of Chagas' disease. AB - Trypanothione reductase from Trypanosoma cruzi is the most promising target molecule for the rational design of a specific drug against Chagas' disease. The recombinant protein was purified in a single chromatographic step and crystallized. Two crystal forms suitable for X-ray diffraction analysis were obtained. Tetragonal crystals (a = b = 87.4 A, c = 152.3 A) were grown from 30% polyethylene glycol (average M(r) = 8,000) in the presence of 0.2% beta-n octylglucoside (space group either P4(2) with one dimer or P4(2)22 with one monomer in the asymmetric unit). Monoclinic crystals (space group P2, a = 136.3 A, b = 91.1 A, c = 126.0 A, beta = 94 degrees) were grown from 1.2 M sodium citrate in the presence of 2% octanoyl-N-methyl-glucamide. They contain two dimers of the enzyme in the asymmetric unit; both crystal forms diffract to 3 A resolution. PMID- 8428619 TI - Ubiquitous presence of chromogranin A in the inner ear of guinea pig. AB - Chromogranin A (CGA), which is supposed to be responsible for the calcium storage of secretory vesicles and is also considered to be a marker protein of neurons and endocrine cells, has been found in a variety of organs and tissues. In the present study, soluble proteins from the organ of Corti, saccule, crista, utricle, tectorial membrane, stria vascularis, and the spiral ligament from the inner ear of guinea pig were extracted, and probed with both polyclonal and monoclonal CGA antibodies to determine the presence of CGA. A 75 kDa protein reactive to both antibodies was found in the organ of Corti, saccule, crista, utricle, stria vascularis, and the spiral ligament, suggesting the widespread presence of CGA in the inner ear. PMID- 8428620 TI - Functional expression of a MAP kinase kinase in COS cells and recognition by an anti-STE7/byr1 antibody. AB - Mitogen-activated protein (MAP) kinases p42mapk and p44mapk are activated by dual tyrosine and threonine phosphorylation in vivo. Both MAPKs are phosphorylated and activated in vitro by an activator recently identified as a protein tyrosine/threonine kinase. We have isolated a putative cDNA for a MAP kinase kinase (MAPKK) and determined its structure [Proc. Natl. Acad. Sci. USA, in press]. The protein encoded by this cDNA shares sequence homology with two yeast protein kinases byr1 and STE7. We now report that stimulation with serum of COS cells expressing this shares sequence homology with two yeast protein kinases byr1 and STE7. We now report that stimulation with serum of COS cells expressing this protein amplifies MAPK activator activity markedly. The increased activity co-migrates during chromatography with the expressed 45 kDa protein, recognized by an anti-STE7/byr1 antibody, and is abrogated by treatment with phosphatase 2A. Thus, this cDNA encodes a functional MAPKK. The anti-STE7/byr1 antibody also recognized a 46 kDa COS cell protein that was resolved from the expressed MAPKK by anion-exchange chromatography. This immunoreactive protein co-eluted with endogenous MAPKK activity, suggesting identification of the immunoreactive band as monkey MAPKK. PMID- 8428621 TI - Characterization of the methylenediphosphonate transport system in Arthrobacter sp. GLP-1 using the novel tritium-labelled derivative. AB - The novel tritium-labelled derivative of methylenediphosphonate (MDP) was used in uptake studies of Arthrobacter sp. GLP-1 capable of utilizing a wide range of organophosphonates as its sole source of phosphorus. The MDP uptake was greatly stimulated upon phosphate deprivation. The uptake obeys Michaelis-Menten kinetics with respective Km and Vmax values of 33 microM and 0.3 nmol.min-1.mg-1 fr.wt. Glyphosate and pyrophosphate were competitive inhibitors of MDP uptake. The effect of orthophosphate was more complex than a mere inhibition of MDP uptake since activation occurred at low concentrations. The uptake of MDP by Arthrobacter sp. strain GLP-1 appears to be mediated by a transport system different from the glyphosate uptake system operating in the same cells. The driving force for MDP uptake by Arthrobacter sp. GLP-1 may be a proton gradient across the cell membrane. PMID- 8428622 TI - Isolation and biochemical characterization of the soluble and membrane forms of folate binding protein expressed in the ovarian carcinoma cell line IGROV1. AB - The human ovarian carcinoma cell line, IGROV1, produces two forms of folate binding protein (FBP), the membrane form that is anchored to the cell surface by a glycosylphosphatidylinositol tail and the soluble form that is shed into the tissue culture medium. Both forms are recognized by the monoclonal antibodies MOv18 and MOv19. Here we describe their purification and biochemical characterization. The purified soluble protein appeared as a single band with an apparent Mr of 36 kDa after SDS-PAGE, whereas the membrane form appeared as a single band with an apparent Mr of 38 kDa. The size difference between the two forms of FBP was confirmed by gel filtration of both the native and the N glycanase-treated proteins. Both purified proteins had equal capacity to bind folic acid. The immunological cross-reactivity and the folic acid binding capability of the FBPs extracted from IGROV1 gave more evidence of the possible existence of a precursor-product relationship between them. PMID- 8428623 TI - Regulation of expression of beta-galactoside alpha 2,6-sialyltransferase in a rat tumor, Zajdela ascitic hepatoma. AB - Tumor cell surface sialic acid levels determine a number of important properties governing cellular interactions and cell-cell communication. Towards understanding the mechanism of regulation of sialic acid levels upon cellular transformation, we have studied the regulation of expression of beta-galactoside alpha 2,6-sialyltransferase in a rat tumor, the Zajdela ascitic hepatoma. We demonstrate distinct differences in the regulation of expression of the enzyme in the tumor cells as compared to normal liver cells. The expression of sialyltransferase is regulated both at the transcriptional and post transcriptional level in a tissue-specific manner. PMID- 8428624 TI - Characterisation of wild-type and mutant forms of human monoamine oxidase A and B expressed in a mammalian cell line. AB - Monoamine oxidase (MAO)-A and MAO-B are FAD-containing mitochondrial enzymes which catabolize biogenic and xenobiotic amines. The N-terminal regions of both forms of MAO contain an ADP-binding consensus sequence found in several dinucleotide-dependent enzymes, but otherwise show remarkable sequence differences. In order to investigate whether the N-terminal region of MAOs participates in the different catalytic properties and inhibitor specificities exhibited by MAO-A and MAO-B, we constructed chimeric A/B forms and expressed them in a human embryonic kidney cell line (293 cells). The MAO-A chimeric form containing the N-terminus (36 amino acids) of MAO-B and the B chimera having the first 45 amino acid sequence of MAO-A were both catalytically active. Compared to the respective wild-type form, they did not show any significant difference in their catalytic properties (Km, kcat) towards the substrates tested or in their sensitivity towards inhibitors. This indicates that the N-terminal region of the two isoenzymes is not involved in the different specificities of MAO-A and MAO-B. Substitution of Cys-397 of MAO-B, i.e. the residue covalently anchoring FAD, with an Ala or a His residue resulted in the total loss of enzymatic activity, suggesting that the covalent coupling of FAD to MAO occurs specifically at the-SH group of cysteine. PMID- 8428625 TI - The low-temperature folding intermediate of hyperthermophilic D-glyceraldehyde-3 phosphate dehydrogenase from Thermotoga maritima shows a native-like cooperative unfolding transition. AB - Hyperthermophilic D-glyceraldehyde-3-phosphate dehydrogenase from Thermotoga maritima, after denaturation in 6 M guanidinium chloride and subsequent renaturation by dilution at 3 degrees C, forms a 'low-temperature intermediate' with its native quaternary structure and most of its dichroic absorption restored, but with significant differences in its fluorescence properties compared to those of the native enzyme. Shifting the temperature beyond 10 degrees C, the enzyme is reconstituted to high yields and to an overall structure indistinguishable from the initial native state [FEBS Lett. 290 (1991) 235-238]. These criteria suggest that the cold intermediate represents an 'assembled molten globule'. However, present equilibrium transition data prove the cold intermediate to be native-like, in that it exhibits a reversible highly cooperative conformational transition to the unfolded state which is incompatible with the typical characteristics of the molten globule state of globular proteins. PMID- 8428626 TI - Interaction between a membrane-associated serine proteinase of U-937 monocytes and peptides from the V3 loop of the human immunodeficiency virus type 1 (HIV-1) gp120 envelope glycoprotein. AB - A trypsin-like proteinase which is inhibited by recombinant gp120 and by synthetic peptides of various lengths spanning the conserved sequence of the V3 loop has been purified and partially characterized from a U-937 cell membrane extract. V3 loop peptides behave as competitive inhibitors of the enzyme, while gp120 exerts a tight-binding inhibition, reacting in stoichiometric amounts with the proteinase to provide significant inhibition. Though the properties of the U 937 membrane proteinase towards gp120 and synthetic peptides of the V3 loop resemble those of the Molt-4 T-cell tryptase TL2, these two proteinases differ by their physicochemical properties and their susceptibility to other inhibitors of serine proteinases. These results give support to the concept of a membrane associated proteinase as a complementary or alternative receptor to the CD4, for allowing virus to enter host cells and thus spreading HIV infection. PMID- 8428627 TI - F0 and F1 parts of ATP synthases from Clostridium thermoautotrophicum and Escherichia coli are not functionally compatible. AB - F1-stripped membrane vesicles from Clostridium thermoautotrophicum and Escherichia coli were reconstituted with F1-ATPases from both bacteria. Reconstituted F1F0-ATPase complexes were catalytically active, i.e. capable of hydrolyzing ATP. Homologous-type ATPase complexes having F0 and F1 parts of ATP synthases from the same origin were DCCD sensitive and supported ATP-driven enhancement of anilinonaphthalene sulfonate (ANS) fluorescence. Hybrid-type ATPase complexes having F0 and F1 parts of ATP synthases from different origins were neither DCCD sensitive nor did they support ATP-driven enhancement of ANS fluorescence. Analyzing these results it has been demonstrated that the F0 and F1 parts of ATP synthases of these two bacteria are not functionally compatible. PMID- 8428628 TI - Induction of metallothionein in a human astrocytoma cell line by interleukin-1 and heavy metals. AB - The effects of cytokines and heavy metals on the expression and localization of metallothioneins (MTs) within U373MG astrocytoma cells were analyzed by using indirect immunofluorescence using a monoclonal anti-MT antibody (MT45). IL-1, CdCl2 (50 microM) or ZnCl2 (500 microM) remarkably augmented intracellular MT levels, whereas IL-6 or 10 microM of ZnCl2 showed no inducing activity. From 24 to 48 h after the addition of CdCl2 or IL-1, immunoreactive MTs were found in the cytoplasm and the nucleus. After 72 h, immunoreactive MTs accumulated in a granular form near the cell surfaces in the presence of CdCl2 (50 microM) or IL-1 plus ZnCl2 (10 microM). However, this accumulation was not observed when only IL 1 was added. Thus, Zn2+ facilitated the appearance of the granular form of immunoreactive MTs at a concentration where they do not induce MTs by themselves. PMID- 8428629 TI - The point mutation of mitochondrial DNA characteristic for MERRF disease is found also in healthy people of different ages. AB - The A-to-G transition mutation in the tRNA(Lys) gene of mitochondrial DNA (mtDNA), characteristic for the maternally inherited MERRF syndrome (myoclonic epilepsy with ragged red fibers), has been identified by point mutation-specific polymerase chain reaction in extraocular muscle from 11 of 16 healthy people of different ages. No mutation was found in navel-string samples from 5 newborns, in HeLa cells, and in 2 individuals younger than 20 years. On the other hand, the mutation is present in all 5 tested 74-89-year-old individuals and in 6 of 9 20 70-year-old individuals. The amount of mutated from total mtDNA was estimated by 'mispairing PCR' in extraocular muscle of 2 individuals of 74 and 89 years to 2.0 and 2.4%, respectively. In most tissue samples the MERRF mutation occurs together with the 'common deletion' of mtDNA, which was previously shown to accumulate in healthy individuals with increasing age. It is proposed that during aging, deletions and point mutations of mtDNA accumulate, which could impair mitochondrial energetics. PMID- 8428630 TI - A D-antifreeze polypeptide displays the same activity as its natural L enantiomer. AB - The D- and L-forms of an alpha-helical antifreeze polypeptide (AFP) have been chemically synthesized. Circular dichroism spectra of the molecules show equal and opposite ellipticites. The D- and the L-enantiomers alone, and a 50:50 mixture of the two, all show identical antifreeze activity, but the enantiomeric forms are predicted to bind to the ice surface with different orientations. It is suggested that symmetry properties of certain ice surfaces permit the asymmetric binding of AFPs, and thus that AFPs are analogous to enzymes that act upon prochiral substrates. PMID- 8428631 TI - Cloning and sequencing of glutamate mutase component E from Clostridium tetanomorphum. Organization of the mut genes. AB - The gene encoding component E, the large subunit, of adenosylcobalamin (coenzyme B12)-dependent glutamate mutase from Clostridium tetanomorphum has been cloned and sequenced. The mutE gene encodes a protein of 485 amino acid residues, with M(r) 53,708. The mutE gene is situated some 1,400 bp downstream of the mutS gene, which encodes the small subunit of glutamate mutase. Between the two is an open reading frame encoding a protein of 462 amino acids, with M(r) 50,171, and of unknown function. All three genes appear to be transcribed as an operon and lie immediately upstream of the gene encoding beta-methylaspartase, the next enzyme in the pathway of glutamate fermentation. Local homology exists between mutE and a region of beta-methylaspartase which contains an active-site serine residue. PMID- 8428632 TI - Binding of alanine-substituted peptides to the MHC class I protein, Kd. AB - Peptides eluted from the native MHC class I molecule, Kd, are generally nonamers that display a strong preference for Tyr in position 2. We investigated the molecular basis for this 'consensus motif' by synthesizing a virally derived peptide, NP 147-155, that is known to be presented by Kd on living cells, and peptide variants of NP 147-155 in which the amino acids in the different positions were sequentially replaced by Ala. All of the peptides bound to purified Kd molecules in vitro with high affinity, except for the peptide in which Tyr2 was replaced by Ala, for which the affinity for Kd decreased at least 100-fold. These results confirm the interpretation of the in vivo studies; namely, that Tyr2 is a critical anchor residue for binding to Kd. PMID- 8428633 TI - Cloning and nucleotide sequence of cDNA for rhodopsin of the squid Todarodes pacificus. AB - A cDNA for rhodopsin was isolated from a library constructed from poly(A)+RNA of the squid (Todarodes pacificus) retina. One positive clone with the longest insert of cDNA (3.1 kb) was selected by employing a PCR-amplified cDNA fragment as a probe. The nucleotide sequence of the cDNA revealed a single open reading frame of 1,344 bp encoding a polypeptide (M(r)49,833), which covered a complete sequence for the squid opsin. This clone had a very long 3'-non-coding region (1.7 kb) including multiple polyadenylation signals, AATAAA, resembling the clones for Todarodes retinochrome and retinal-binding protein (RALBP). The analysis of hydropathicity demonstrated the presence of seven transmembrane spanning domains, and a possible retinal-binding site, Lys-305, was found in the 7th domain. Todarodes rhodopsin contained characteristic sequences of PPQGY repeated in the C-terminal region, as reported in Loligo and octopus rhodopsins. Structural comparison of those cephalopod rhodopsins is also discussed. PMID- 8428634 TI - Association of three small GTP-binding proteins with cholinergic synaptic vesicles. AB - Several small (low molecular weight) GTP-binding proteins are associated with cholinergic synaptic vesicles derived from the electric organ of electric ray. Using GTP overlay techniques and direct micro sequencing we analyzed the association of small GTP-binding proteins with synaptic vesicles. Both experimental procedures revealed the specific occurrence of multiple small GTP binding proteins with this organelle. Moreover, direct amino acid sequence analysis assigned at least three different small GTP-binding proteins, ora3, o ral and o-rab3, to the vesicular compartment. Furthermore, the data reflect the relative abundance of these three proteins on the vesicle membrane, thereby demonstrating the predominant occurrence of o-rab3, the only exclusively synaptic vesicle specific small GTP-binding protein. PMID- 8428635 TI - ATP-dependent protein synthesis in isolated pea chloroplasts. Evidence for accumulation of a translation intermediate of the D1 protein. AB - In the presence of externally added ATP, in the dark, isolated pea chloroplasts accumulate two proteins of molecular masses of about 22 and 24 kDa which precipitate with specific antibodies raised against the D1 protein. By chasing in the light, these proteins disappeared on the fluorogram concomitant with the appearance of the precursor- and mature-sized D1 proteins. Polysome analysis indicated that the 22-kDa component is associated with membrane-bound ribosomes and is thus ascribed to a translation intermediate of the D1 protein. On the other hand, the 24-kDa component could not be found in the polysome fraction under the experimental condition used, suggesting the possibility that this component is a degradation product of the D1 protein. The conclusion from this analysis is that the synthesis and/or stable accumulation of the D1 protein requires factor(s) caused by illumination, in addition to ATP, in isolated pea chloroplasts. PMID- 8428636 TI - Two 3',5'-cyclic-adenosine monophosphate response elements in the promoter region of the human gastric inhibitory polypeptide gene. AB - Transfection of chimeric chloramphenicol acetyltransferase plasmids containing various deletions of the human gastric inhibitory polypeptide (GIP) promoter into hamster insulinoma (HIT T15) cells indicated that the region between -180 and +14 is sufficient for basal promoter activity. Two CRE-BP1 binding sites were identified in this promoter region by DNase I footprinting with the bacterially expressed cAMP response element (CRE) binding protein, CRE-BP1. Mutation analyses showed that these two CREs are required for the basal promoter activity, and furthermore that one site, at nucleotide-158, contributed mainly to the cAMP inducibility of the GIP promoter in HIT T15 cells. Interestingly, the GIP promoter activity was repressed by the c-jun proto-oncogene product, possibly through the CREs. PMID- 8428637 TI - Regulation of cardiac insulin receptor function by guanosine nucleotides. AB - The present study examined the effect of GTP-gamma-S on the function of insulin receptors partially purified from adult rat cardiomyocytes by WGA chromatography. GTP-gamma-S increased receptor autophosphorylation about two times and fully mimicked the stimulatory action of insulin on poly(Glu:Tyr) phosphorylation with no additional effect of the hormone. The effect of GTP-gamma-S was specific, dose dependent, and due to an increase in the Vmax of the kinase. In the presence of ATP or AMP-PNP, insulin significantly enhanced the binding of [35S]GTP-gamma-S to the partially purified insulin receptor. The findings suggest coupling of the insulin receptor to a G-protein which may be involved in the regulation of tyrosine kinase activity. PMID- 8428638 TI - Stabilization of Bacillus stearothermophilus neutral protease by introduction of prolines. AB - The thermostability of neutral proteases has been shown to depend on autolysis which presumably occurs in flexible regions of the protein. In an attempt to rigidify such a region in the neutral protease of Bacillus stearothermophilus, residues in the solvent-exposed 63-69 loop were replaced by proline. The mutations caused large positive (Ser-65-->Pro, Ala-69-->Pro) or negative (Thr-63- >Pro, Tyr-66-->Pro) changes in thermostability, which were explained on the basis of molecular modelling of the mutant proteins. The data show that the introduction of prolines at carefully selected positions in the protein can be a powerful method for stabilization. PMID- 8428639 TI - Augmentation of human natural killer cells by splenopentin analogs. AB - Splenopentin, Arg-Lys-Glu-Val-Tyr (SP-5) and its synthetic analogs; Arg-D-Lys-Glu Val-Tyr (pentapeptide 1), Lys-Lys-Glu-Val-Tyr (2), D-Lys-Lys-Glu-Val-Tyr (3), Arg Lys-Gly-Val-Tyr (4), and Arg-Lys-Gln-Val-Tyr (5) have been examined for augmentation of human natural killer (NK) cell activity and human T-cell transformation response. Pentapeptides 2 and 3 were found to significantly augment the in vitro human NK cell activity. However, none of them had any effect on lymphocyte proliferative responses. PMID- 8428640 TI - Structure of the 16 S ribosomal RNA of the thermophilic cyanobacterium Chlorogloeopsis HTF ('Mastigocladus laminosus HTF') strain PCC7518, and phylogenetic analysis. AB - The thermophilic cyanobacterial strain, PCC7518, originally identified as 'Mastigocladus laminosus HTF' does not show branchings or heterocysts. The absence of branchings supports the later assignment to the genus Chlorogloeopsis. The absence of heterocysts may be the result of a mutation because heterocysts were observed in the original isolate. Alternatively, contamination may have happened. To solve this problem, the 16 S rRNA sequence was determined and used to infer a secondary structure model and build distance trees. The trees showed that strain PCC7518 belongs to the cluster of heterocystous species and has most probably lost the ability to produce heterocysts by mutation. It is only distantly related to Chlorogloeopsis fritschii PCC6718. PMID- 8428641 TI - Melatonin secretion in vitro from the pineal complex of the lamprey Petromyzon marinus. AB - Melatonin secretion from cultured pineal complexes of the lamprey, Petromyzon marinus, was measured by radioimmunoassay (RIA) for 6 days under either a 12:12 light:dark cycle (L:D) or continuous darkness (D:D) at either 10 degrees or 20 degrees. Under the L:D cycle, melatonin release was completely suppressed during daytime and high during night-time, with larger amplitude at 20 degrees than at 10 degrees. Under D:D conditions, no significant circadian rhythmicity in secretion could be revealed by periodogram analysis at either 10 degrees or 20 degrees. This is the first direct evidence of melatonin secretion in a lamprey, suggesting that both light and temperature can affect in vitro melatonin secretion. PMID- 8428642 TI - The significance of olfaction and social cues in milt availability, sexual hormone status, and spawning behavior of male rainbow trout (Oncorhynchus mykiss). AB - Male rainbow trout orientate to a "releaser" pheromone emitted by an ovulated female. This chemical signal is not necessary for the completion of spawning behavior: anosmic males spawned as readily as intact males when paired with ovulated females. The amount of "strippable" milt and plasma concentrations of testosterone (T) and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) were greater in males paired for 3 hr with ovulated females than in isolated males, or males placed with nonvitellogenic adult females or in all male groups. Milt and plasma T, 17,20 beta-P, and 11-ketotestosterone (11-KT) were reduced in anosmic males paired with ovulated females, indicating the existence of a priming pheromone. Milt, T, 11-KT, and 17,20 beta-P increased in intact males within 4 hr after injection of salmon gonadotrophin-releasing hormone analogue, lyophilized salmon pituitary extract, or homogenized fresh rainbow trout pituitary material. The results indicate that the priming pheromone acts through the hypothalamus pituitary-gonad axis. PMID- 8428643 TI - Thyrotropin-releasing hormone (TRH) is the major prolactin-releasing factor in the bullfrog hypothalamus. AB - A substance exhibiting potent activity in stimulating the release of prolactin from bullfrog (Rana catesbeiana) pituitary in vitro was isolated from an acid extract of bullfrog hypothalami by gel-filtration chromatography (Sephadex G-15), ion-exchange chromatography (Mono-S HR 5/5), and reverse-phase high-performance liquid chromatography (TSK-gel ODS-120T). Its amino acid composition was similar to that of synthetic thyrotropin-releasing hormone (TRH). Radioimmunoassay confirmed that the substance had TRH immunoreactivity. Moreover, it exhibited the same chromatographic behavior as that of synthetic TRH. These results clearly indicate that the isolated hypothalamic substance is TRH, and that it is the major prolactin-releasing factor present in the bullfrog hypothalamus. PMID- 8428644 TI - Comparative studies of thyroxine binding proteins in turtle plasma using column chromatography. AB - Thyroxine (T4) binding proteins in turtle plasma were compared with those of various mammalian and nonmammalian (fish, birds) species using gel filtration chromatography (Bio-Gel). High-affinity T4 binding was observed in all mammals studied, in the chicken, and in one species of turtle, Trachemys scripta. This high affinity T4 binding protein (TBP) appears to be the major component of T4 transport in the turtle; selective removal of this TBP greatly diminished binding activity in T. scripta plasma. In all other nonmammalian species studied (including other turtles), T4 binding appeared to be associated primarily with relatively low affinity components such as albumin. Albumin concentrations in turtle (T. scripta, and Chelydra serpentina) plasmas were examined by measurement of dry weight, protein assay (Bradford), radioimmunoassay, and electrophoretic densitometry. While C. serpentina may have total protein concentrations approaching that of human plasma, albumin content in both turtles was estimated at about 3-4 mg/ml, less than 10% that of human plasma albumin; larger molecular weight components that do not bind T4 comprise a greater proportion of turtle plasma protein. Additionally, purified T. scripta albumin exhibits lower T4 binding than does human albumin; binding to turtle albumin is also reduced by fatty acids. Taken together, these results indicate a lesser role for albumin in plasma T4 binding in T. scripta than in humans, and, thus, a larger role for the T4 binding protein, TBP. Other turtles lacking a TBP may rely wholly on low affinity albumin binding that also has a lower capacity than that in mammals. PMID- 8428645 TI - Inhibition of in vitro pituitary gonadotropin secretion by 17 beta-estradiol in the frog Rana pipiens. AB - We tested the effects of low concentrations of 17 beta-estradiol (E2) on the in vitro pituitary responsiveness to gonadotropin-releasing hormone (GnRH) in frogs (Rana pipiens). We first showed that the pH indicator phenosulfonpthalein (phenol red, PR) at levels normally used in the culture medium did not significantly affect pituitary basal or GnRH-stimulated gonadotropin secretion in male glands, as may occur in mammals. Incubation of male glands with E2 at 10 pg/ml for 24 hr in medium lacking PR did not alter basal follicle-stimulating hormone (FSH) or luteinizing hormone (LH) secretion, but significantly attenuated responsiveness of both gonadotropins to GnRH (0.2, 1, and 5 ng/ml); E2 did not affect responses at the highest dose of GnRH (25 ng/ml). GnRH responsiveness of female pituitaries was also inhibited by E2 at all doses of GnRH; 1 ng/ml E2 had greater effects than did 10 pg/ml. The present results demonstrate that E2 at concentrations commonly found in the circulation of both sexes have a consistent direct inhibitory effect on FSH and LH secretion at the level of the pituitary. PMID- 8428646 TI - Effects of acclimation to hypertonic environment on plasma and pituitary levels of two prolactins and growth hormone in two species of tilapia, Oreochromis mossambicus and Oreochromis niloticus. AB - Specific radioimmunoassays (RIAs) for the pair of tilapia prolactins (tPRLs) and growth hormone (tGH) were developed using antisera raised in rabbits. Anti tPRL177 did not cross-react with tPRL188 and tGH. Anti-tPRL188 did not cross react with tPRL177 and showed slight cross-reaction (3.1%) with tGH. Anti-tGH showed negligible cross-reactions with tPRL177 (0.4%) and tPRL188 (1.6%). Pituitary homogenates and plasma from Oreochromis niloticus exhibited displacement curves parallel to the standards in the three RIAs. Plasma from hypophysectomized O. niloticus showed no cross-reaction in any of the three RIAs. Plasma and pituitary levels of the two PRLs in O. mossambicus in freshwater did not differ significantly from each other, whereas in O. niloticus, the levels of PRL177 were significantly greater than those of PRL188 in both plasma and pituitary. After acclimation for 3-4 weeks in seawater (O. mossambicus) or 50% seawater (O. niloticus), the levels of both PRLs decreased significantly compared to their levels in freshwater. Acclimation to a hypertonic environment did not affect plasma and pituitary GH levels in either species. Immunocytochemical staining of the pituitary of O. niloticus revealed colocalization of both PRLs in rostral pars distalis. Our findings suggest that the synthesis and secretion of the two tPRLs could be independently regulated in the same cells. PMID- 8428647 TI - Photoperiodic control of annual reproductive cycle in subtropical brahminy myna, Sturnus pagodarum. AB - The aims of this investigation were to: (i) study the effect of the length of photoperiod on the timing of onset of gonadal recrudescence and postreproductive decline in photoresponsivity, and (ii) identify the mechanism(s) involved in the initiation of gonadal recrudescence in the subtropical brahminy myna (Sturnus pagodarum). Two series of experiments were performed using adult birds: (1) Groups of birds were subjected to 12L:12D, 14L:10D, and 16L:8D in January and to 12L:12D and 14L:10D in March. Those exposed to 12L and 14L in March were transferred to longer daylengths (16L:8D, 20L:4D) after 180 days. In the January birds, 14L and 16L induced testicular growth and regression earlier than 12L; also, 16L caused gonadal recrudescence earlier than 14L. The 14L photoperiod, however, induced gonadal development in March birds within 30 days, similar to that found in January birds under 16L:8D. The response of March birds to 12L:12D was, however, similar to those of January birds, except that the time of maximum testicular response and subsequent regression was advanced in the former group. It is concluded that the refractoriness to long day photostimulation is fully dissipated in March but not in January. (2) Groups of unstimulated and stimulated birds were exposed to "resonance" and "interrupted-night" light: dark cycles for 5 weeks. Resonance LD cycles of 12 (8L:4D)- and 36 (8L:28D)-hour induced day responses, while groups under 24 (8L:16D)- and 48 (8L:40D)-hr cycles behaved as if exposed to short days. In night-interruption experiments, a 1 hr light pulse given 12 and 16 hr, but not 20 hr, after dawn induced the long day response. These results suggest that in brahminy myna the time to the onset of gonadal recrudescence and subsequent regression (photorefractoriness) is dependent on the length of photophase as well as time of the year when exposed to stimulatory photoperiods, and that a light-sensitive rhythm with a period of about 24 hr is involved in the photoperiodic induction of gonadal recrudescence. PMID- 8428648 TI - Estrogen enhancement of Ca-, Mg-, and Ca-Mg-stimulated adenosine triphosphatase activity in the chick shell gland. AB - The effect of 17 beta-estradiol (E2) on Ca-, Mg-, and Ca-Mg-ATPase activity was investigated in the shell gland of 6-week-old chicks. In the first study, each of 42 birds was implanted with three E2 (Compudose-200) pellets. An additional 6 sham-implanted birds served as controls for measurements of body weight and concentrations of E2 and Ca in plasma. The activities of Mg-, Ca-, and Ca-Mg ATPase peaked coincident with maximum plasma E2 concentrations 8 days after implantation and then progressively declined in concert with the decline in plasma E2. By 29 days after implantation, the ATPase activities were similar to those measured in birds whose E2 pellets had been removed for 11 days. Concentrations of plasma E2 dropped from peak values of 1676 +/- 317 at Day 8 to 611 +/- 180 pg/ml at Day 29. When birds whose E2 pellets had been removed were reimplanted with three pellets per bird, plasma E2 again increased to 1637 +/- 227 pg/ml. ATPase activity in these reimplanted birds also was greater (P < 0.05) than activities measured in E2-removed or E2-maintained birds. In a second study the Ca-ATPase Km and Vmax were determined in E2-implanted chicks (three pellets per bird) and compared to E2-withdrawn chicks. Five days after reimplantation of chicks with E2, there was a significant increase in both Vmax (3.38 +/- 0.21 vs 2.37 +/- 0.28 micrograms Pi/mg protein/min; P < 0.05) and Km (0.31 +/- 0.02 vs 0.25 +/- 0.01 mM Ca; P < .01).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428650 TI - Localization of mRNAs for a pair of prolactins and growth hormone in the Tilapia pituitary using in situ hybridization with oligonucleotide probes. AB - Oligonucleotide probes were synthesized for the mRNAs of a pair of tilapia prolactins (tPRL177 and tPRL188) and growth hormone (tGH) based on cDNAs for the hormones of Oreochromis niloticus and amino acid sequences for the hormones of O. mossambicus. The three 45mer probes were labeled with 35S for hybridization studies on pituitary sections of O. mossambicus adapted to fresh water (FW) or seawater (SW). Expression of tPRL mRNA in the rostral pars distalis was clearly evident with either PRL probe in adjacent sections in PRL cells of the rostral pars distalis; mRNAs of both PRLs were colocalized in the same cells. In addition, the tGH probe demonstrated expression of tGH mRNA specifically in GH cells in the proximal pars distalis. The hybridization signals for both PRLs were significantly greater in the rostral pars distalis of FW fish than in that of SW fish, as judged by computer-aided analysis. In addition, grain concentration for both PRLs was significantly greater over centrally located PRL cells of FW fish. In addition, although overall grain concentrations were lower in SW fish, there were significantly more grains over the centrally located PRL cells with the tPRL177 probe, whereas there was no difference with the tPRL188 probe. There was no detectable difference in the occurrence of tGH mRNA between FW and SW fish. PMID- 8428649 TI - Recombinant carp (Cyprinus carpio) growth hormone: expression, purification, and determination of biological activity in vitro and in vivo. AB - Carp growth hormone (cGH) cDNA (Koren et al., 1989) was cloned under the control of lambda-phage PLOL promoter and lambda cll ribosomal binding site into pBR322 plasmid to enable its expression in Escherichia coli A1645 that produces constitutively the thermolabile lambda repressor c1857. Temperature shift to 42 degrees abolished the repression, resulting in a high level of cGH expression. The bacterially expressed cGH protein, contained within the refractile body pellet, was solubilized in 4.5 M urea, refolded, and purified on Q-Sepharose column by stepwise elution with NaCl. The bioactive fraction was eluted at 0.2 M NaCl at a yield of 10-15%. This fraction contained predominantly (95%) 21.5-kDa monomeric cGH. The activity of cGH in vitro was bioassayed using Nb2-11C lymphoma cells (containing lactogenic receptors) and 3T3-F442A preadipocyte cells (containing somatogenic receptors). Bioactivity was found to be 0.01 and 6-10% that of human GH, respectively. In vivo cGH activity was measured by weekly ip injection in juvenile carp fed a low (23%) protein diet. Over a 6-week period, cGH increased the growth rate by 38% compared to fish injected with vehicle only. Identical injections with bovine GH yielded only a 21% increase. PMID- 8428651 TI - Epidermal growth factor-like immunoreactivity in the buccopharyngeal mucous glands of Xenopus laevis tadpoles. AB - We report epidermal growth factor (EGF)-like and epidermal growth factor receptor (EGFR)-like immunoreactivity in the buccopharyngeal mucous glands of Xenopus laevis larvae. EGF-like immunostaining was heaviest at the apices of the secretory cells of these glands. Immunostaining for EGFR-like protein was also observed in the mucosal lining of the alimentary tract. This staining was heaviest in cells lining the foregut but was almost absent in cells from the hindgut. A potential role for an orally secreted EGF homologue in anuran amphibians is discussed. Wassersug (1986) hypothesized the existence of a metamorphic inhibitory agent, produced by larval buccopharyngeal mucous glands, which could indirectly link food ingestion to the endocrine control of metamorphosis. The presence of EGF-like immunoreactivity in the oral mucous glands of X. laevis larvae, as well as the presence of EGFR-like immunoreactivity in the gut wall of this tadpole, satisfies many of the criteria for Wassersug's regulatory agent. It remains to be shown that EGF (or an anuran EGF homologue) has a direct inhibitory effect on anuran metamorphosis when administered via an orogastric route. PMID- 8428652 TI - Excess iodide and thyroid function in the developing chick embryo. AB - The effects of adding excess iodide (I-) to egg yolks were studied on the quantity of iodide transferred to the chick embryo and on thyroid function from Days 8 to 18 of incubation. Following injection of either 0.5 or 1 mg of iodide into the yolk on the second day of incubation, embryonic plasma iodine levels increased significantly over levels of controls at the same age. These increases were proportional to the amount of excess iodide in the yolk. Moreover, a constant ratio between the iodine levels of the allantoic fluid and the plasma was maintained for any given day of incubation. Up to Day 10, the iodine content of the embryonic thyroid was closely correlated with the increased plasma iodine concentrations. After Day 10, concomitant with the establishment of the hypothalamo-adenohypophyseal-thyroid axis, the increase in thyroid iodine no longer paralleled that of plasma iodine, but was only twice that of controls. As thyroid weight doubled in these treated embryos, thyroid iodine concentrations of control and treated embryos were similar. Iodide excess did not block thyroid iodine organification, nor was there any modification of iodine distribution among the different iodo-amino acids. Moreover, despite the doubling of thyroid hormone content in the goiters of treated embryos, circulating levels of thyroid hormones in control and treated embryos were not different. PMID- 8428653 TI - Cloning and sequencing of potato virus Y (Hungarian isolate) genomic RNA. AB - A sequence of 9703 nucleotides (nt) is reported for the genomic RNA of potato virus Y (Hungarian isolate, PVY-H), which causes necrotic rings around the buds on the tubers and mottling of leaves. The sequence contains one large open reading frame of 3061 amino acids (aa), a noncoding region of 189 nt at the 5' end and a 330-nt 3' nontranslated region. The nt sequence and the predicted aa sequence of the polyprotein of PVY-H were analysed pairwise with the only available complete sequence of PVY strain N (PVYn) and with the partial sequences of different PVY strains, as well as with other potyviruses and potyvirus-related plant viruses. The overall relationship between PVY-H and PVYn shows a nt sequence identity of 88.5% and an aa sequence identity of 94.2%. The lowest degree of homology was detected at the 5' terminus of the genome, including the 5' noncoding region (70.3%) and the 275-aa P1 protein (78%). A fivefold sequence repeat block of 5'-UUUCA was found in the 5' noncoding region of PVY-H, which seems to be characteristic of PVY strains. PMID- 8428654 TI - Isolation of a tomato alcohol dehydrogenase 2-encoding cDNA using phage-promoted antibody screening of a plasmid cDNA library. AB - We describe the cloning of a cDNA encoding tomato alcohol dehydrogenase 2 (Adh2) by screening plasmid cDNA clones in phage plaques. A cDNA library constructed in a plasmid vector containing a unique SstI site at the 5' end of the cDNA insert was transferred into the SstI site of the lacZ gene of phage lambda Charon16, and screened by anti-Adh2 antibody to identify reactive plaques. Plasmid cDNA clones were recovered by SstI digestion, ligation, and transformation from phage minipreps for subsequent characterization. This system preserves the original plasmid library for subsequent screening with nucleic acid probes to identify full-length, multiple independent, or related cDNA clones not subject to the selection pressure of phage growth or lysogeny, or negative antibody reactivity. Thirty-two cDNA clones were identified with polyclonal antiserum to Adh2. Three of these reacted with monoclonal anti-Adh2 and only those three hybridized to maize adh1 sequence. One of these cDNAs, Adh31, was further characterized as encoding Adh2 by hybrid-selected translation and high sequence homology with the maize adh1 gene. PMID- 8428655 TI - Cloning, expression and sequences of mouse sterol-carrier protein-x-encoding cDNAs and a related pseudogene. AB - The expression of the sterol-carrier protein 2 (SCP-2)-encoding gene (SCP-2) is unusually complex. At least four SCP-2-related transcripts are detected in mouse liver: two, of 1.6 and 3.0 kb, are expressed to high levels while the other two, of 0.9 and 2.2 kb, reveal relatively low expression. Hybridization with a probe which specifically hybridizes with the rat SCP-2-related cDNA encoding rat SCP-x reveals that the 2.2- and 3.0-kb transcripts encode mouse SCP-x. SCP-x transcripts are expressed predominantly in the liver, but low-level expression can be demonstrated in all tissues analyzed. Isolation and characterization of two overlapping SCP-x cDNAs indicate that the cDNAs are derived from alternatively polyadenylated transcripts spanning approx. 2.2 and approx. 2.9 kb. Nucleotide sequencing reveals that the predicted ORF, which consists of 547 codons, is composed of 143 C-terminal amino acids which are essentially identical with mouse pre-SCP-2 and 404 N-terminal residues which are specific for SCP-x. To date, it is not clear if all SCP-2-related transcripts are transcribed from a single gene. We have isolated a genomic clone containing an SCP-2-related pseudogene which has some of the characteristics expected for a truncated processed pseudogene. Therefore, our results indicate that at least some of the multiple restriction fragments which are detected by Southern hybridization analyses with SCP-x cDNA-derived probes can be explained by cross-hybridization with a pseudogene. PMID- 8428656 TI - Sequences and expression of the porcine apolipoprotein A-I and C-III mRNAs. AB - Apolipoprotein A-I (ApoA-I) is the principal protein component of plasma high density lipoprotein (HDL) and an activator of lecithin:cholesterol acyltransferase. Apolipoprotein C-III (ApoC-III) exchanges between triglyceride rich lipoproteins and HDL and inhibits the lipolysis and uptake of triglyceride rich lipoproteins. To study the expression of these Apo-encoding genes in the developing swine, apoA-I and apoC-III cDNAs from a lambda gt11 porcine liver cDNA library and apoC-III from a porcine genomic DNA library were isolated and sequenced. The predicted amino acid (aa) sequence and composition for ApoC-III matched the N-terminal aa sequence and composition of purified swine ApoC-III. Comparison among known ApoA-I and C-III aa sequences from various species revealed strict conservation of amphipathic helices. In adult pigs, the apoA-I mRNA was found predominantly in the intestine and liver, with a small amount detected in the testes. In contrast, apoC-III mRNA was found predominantly in adult liver. Developmentally, hepatic apoA-I and apoC-III mRNAs were expressed in livers of fetal, newborn, and suckling animals. Intestinal apoA-I and apoC-III mRNAs, however, were detected only in postpartum animals. Although intestinal apoA-I mRNA expression continued into the adult, intestinal apoC-III mRNA expression declined sharply after the newborn period. PMID- 8428657 TI - Cloning, sequencing and expression of a cDNA encoding mammalian valyl-tRNA synthetase. AB - A fragment of the cDNA encoding a rat valyl-tRNA synthetase (TrsVal)-like protein was cloned from a rat cDNA library in lambda gt11 using an oligodeoxyribonucleotide (oligo) probe. Three independent plaque clones containing the human TrsVal cDNA were then isolated from a lambda gt10 human erythroleukemia cDNA library using the rat cDNA fragment as the hybridization probe. Sequence analyses of the cDNA fragments provided a 3.2-kb sequence with an open reading frame that contained the 'HIGH' synthetase signature sequence and the tRNA 3'-end-binding motif, KMSKS, and putative Val-binding motif, EWCISRQ. The sequence was extended to the 3' end of the cDNA by the polymerase chain reaction using an internal primer and an oligo(dT) adapter. The deduced 1051 amino-acid sequence shares 65% identity with yeast TrsVal, and contains a highly basic N-terminal region, a newly evolved protease-sensitive region in sequence close to the C terminus, and several sites for protein kinase C phosphorylation. A 3-kb cDNA fragment was sub-cloned into plasmid pSVL and expressed in COS-7 cells; up to a sevenfold increase in TrsVal activity was obtained. These results confirm the cloning and sequencing of a human TrsVal-encoding cDNA. PMID- 8428658 TI - The complete sequence of the gene encoding bovine alpha s2-casein. AB - From a bovine genomic library, five overlapping clones, spanning some 50 kb, have been isolated. These clones contain the complete alpha s2-casein-encoding gene (alpha s2ca) and its 5' and 3' flanking regions. The nucleotide (nt) sequence of the complete gene including 2510 bp of the 5' flanking region and 276 bp of the 3' region has been determined. The total length of alpha s2ca appears to be 18483 bp and, therefore, it is the longest of the four bovine casein-encoding genes. The alpha s2ca gene is comprised of 18 exons ranging in size from 21 to 266 nt. There are 16 Alu-like artiodactyla retroposons inserted at ten different locations within the gene. About 14% of the gene is composed of these repetitive sequences. Although the organization of alpha s2ca appears to be similar to that of the alpha s1-casein-encoding gene (alpha s1ca), sequence comparisons and the length of the exons indicate that it is more closely related to the beta-casein encoding gene. Furthermore, it is shown that both genes could have evolved from a common ancestor by means of internal duplications. PMID- 8428659 TI - Amplification of the gene encoding the alpha-subunit of the mitochondrial ATP synthase complex in a human retinoblastoma cell line. AB - A cDNA clone encoding the precursor of the alpha-subunit of the human mitochondrial ATP synthase (F1-ATPS) complex was isolated from a library prepared from the poly(A)+ RNA present in a retinoblastoma (RB) cell line. Northern blot analysis of RNAs derived from a variety of transformed cell lines as well as from normal human fetal tissues indicated that RNA expression was significantly higher in two of the four RB cell lines analysed, Y79 (10- to 30-fold) and RB522A (3- to 8-fold), than in other cell lines or tissues. The increased mRNA level was apparently the result of gene amplification in Y79, but not in RB522A. PMID- 8428660 TI - Efficient production of biologically active human salivary cystatins in Escherichia coli. AB - Different Escherichia coli expression systems were used for expression of cDNA clones encoding the human salivary cysteine proteinase (CysP) inhibitors, cystatins SN and S (CsnSN and CsnS). These included pOTSNco12 that expresses foreign sequences as authentic (nonfusion) proteins, and pGEX-2T that directs the synthesis of foreign polypeptides as fusion proteins with glutathione S transferase (GST). The pOTS vector produced low levels of recombinant CsnSN (reCsnSN) that was localized in the soluble fraction, but not easily purified. The pGEX vector, on the other hand, produced much higher yields of the fusion protein, GST::CsnSN, that was localized almost entirely in the insoluble protein fraction. Solubilized and refolded GST::CsnSN inhibited the CysP, papain, more efficiently than chicken egg white Csn, indicating that the recombinant product was biologically active and that the GST carrier did not interfere with the biological activity. The pGEX-2T vector was subsequently used for the large-scale production of reCsnSN and reCsnS that were cleaved from the GST by thrombin and purified by DE-52 cellulose chromatography. ReCsnSN inhibited papain almost as efficiently as salivary CsnSN, while the reCsnS showed lower inhibitory activity as compared to both salivary CsnS and reCsnSN. PMID- 8428661 TI - Characterization of an evolutionarily old human alphoid DNA. AB - A recently isolated human alphoid DNA (in plasmid pHH550) has been sequenced and found to have an exceptionally high degree of similarity to the human alphoid consensus sequence, while its component monomers are unusually heterogeneous in sequence. In contrast to other alphoid DNAs, this DNA is found in all primates tested. Thus this may be an evolutionarily old sequence similar to the one from which other human alphoid DNAs diverged. The pHH550 sequences are found on a number of human chromosomes, including 21 and 22. On chromosome 21 most members of this new sequence group are located distal to other alphoid DNAs. PMID- 8428662 TI - Molecular organization and chromosomal location of human GC-rich heterochromatic blocks. AB - From the sequencing of three genomic DNA fragments and PCR amplification products from total human DNA, we have derived the sequence of a 545-bp Sau3A fragment (68% GC), representative of a family of human DNA repeats. Since previous studies suggested its linkage with unrelated Sau3A repeats of 68 bp (54% GC) (beta satellite sequences), this feature was further investigated by in situ hybridization experiments and by Southern blot analysis of a panel of DNAs from human-Chinese hamster somatic cell hybrids. Both DNA repeats are preferentially localized on the heterochromatic regions of acrocentric chromosomes, on the pericentromeric heterochromatin of chromosome 1, 3 and 9, and on the proximal euchromatic region of the chromosome Y q arm. On chromosome 9, both repeats are part of a 2.7-kb higher-order repeat unit. These results and the Southern blot analysis on partial digests of total DNA, suggest that the linkage between the two repetitive DNA sequences is a constant feature throughout the genome. Furthermore, Southern blot analysis of HpaII-digested and MspI-digested DNA from different human tissues and tumor cell lines indicates that the investigated heterochromatic blocks appear to be subjected to changes in their methylation pattern. PMID- 8428663 TI - CACC box and enhancer response of the human embryonic epsilon globin promoter. AB - The functional interaction between the human epsilon globin promoter and an erythroid-specific transcription enhancer, 5' HS-2, has been analyzed by transient expression assay. While stepwise deletion of DNA sequences between -852 and -122 had only small effects, removal of the CACC box at position -111 greatly decreased epsilon-globin promoter activity, as well as its response to the enhancer function of 5' HS-2 in erythroid cells. Our data demonstrated that the three ubiquitous promoter elements, the CACC, CCAAT, and TATA boxes, of the epsilon-globin-encoding gene together form a minimal promoter that would interact efficiently with 5' HS-2, and that at least the CACC box is an essential functional component of this enhancer-promoter interaction. PMID- 8428664 TI - Effect of heat denaturation of target DNA on the PCR amplification. AB - The polymerase chain reaction (PCR) and amplification of specific regions of DNA in vitro is a widely used and powerful technique, and the optimization of conditions used to maximize PCR product yield has received much attention. We have shown that lengthy denaturation times of template DNA ranging from 1 to 7 min at pH 7.0-8.0, that are often employed prior to the start of a PCR reaction, result in marked degradation of the template. This can result in a significant reduction in the yield of PCR products larger than 500 bp, by up to 99%. This effect was demonstrated for both complex genomic template DNA, and also for a 2691-bp linear piece of template DNA using both a rapid hot-air thermocycler and a conventional block thermocycler. This decrease in product yield is likely due to the increased degradation of the template or target DNA as a result of pre amplification denaturation (PAD). We therefore recommend that when amplifying larger pieces of DNA, the template DNA should not be exposed to PAD prior to a PCR reaction, irrespective of the starting pH of the template solution. PMID- 8428665 TI - Analyses of genes that encode the 15-kDa zein protein of maize: identification of potential gene regulatory elements. AB - A gene (gZ15.4.1) encoding the 15-kDa zein polypeptide was isolated from maize cultivar A5707, and its nucleotide sequence was determined. A total of 2085 bp was sequenced, including about 300 bp of 5'-flanking DNA that includes several potential regulatory elements not available in the previously published 15-kDa zein-encoding gene (gZ15A) sequence. Several nt differences between gZ15.4.1 and gZ15A were observed, which include two single amino acid replacements. PMID- 8428666 TI - Identification of two new members of the 1-aminocyclopropane-1-carboxylate synthase-encoding multigene family in mung bean. AB - The key enzyme regulating ethylene biosynthesis in higher plants is 1 aminocyclopropane-1-carboxylate (ACC) synthase. In mung bean (MB), the existence of three genes encoding this enzyme has previously been reported [Botella et al., Plant Mol. Biol. 18 (1992) 793-797], one of which corresponds to a full-length indole-3-acetic acid-inducible cDNA [Botella et al., Plant Mol. Biol. (1992) 425 436]. In this paper we report the cloning of two new genomic sequences coding for ACC synthase in MB (MAC-4 and MAC-5). MAC-4 is 1340 bp long and encodes 388 amino acids (aa) while MAC-5 is 1393 bp long and encodes for 391 aa. Genomic Southern analysis suggests the existence of only one copy of each gene in the genome. PMID- 8428667 TI - The complete nucleotide sequence of cauliflower mosaic virus isolate BBC. AB - We report the complete nucleotide sequence of BBC, a new and unique isolate of cauliflower mosaic virus (CaMV). The organization of the BBC genome agrees with that of previously sequenced CaMV isolates. PMID- 8428668 TI - High-level expression in Escherichia coli of the gene coding for the major structural protein (p72) of African swine fever virus. AB - The gene encoding the major structural protein (p72) of African swine fever virus (ASFV) has been expressed in Escherichia coli using a T7 RNA polymerase system. The use of a recombinant plasmid which contains the entire gene inserted between the T7 promoter and the transcription terminator of the expression vector allowed us to obtain a high expression level of the intact viral protein. This polypeptide, which appears in the insoluble fraction of the bacterial extracts, showed an intense reaction with the antibodies present in the sera of ASFV infected animals, as demonstrated by Western blot and enzyme-linked immunosorbent assay. The recombinant protein was purified by size-exclusion high-performance liquid chromatography and used to develop a serological test of the disease. PMID- 8428669 TI - Isolation and sequence of a mouse brain cDNA coding for protein kinase C-gamma isozyme. AB - The regulatory enzyme, protein kinase C (PKC), is characterized by a family of related isozymes. Currently, nucleotide (nt) sequences for seven members of this family have been reported from the bovine, human and rat genomes. Only four of these seven PKC isoforms have been isolated in mouse: alpha, beta II, delta and epsilon. Here, we report the cDNA sequence encoding mouse PKC-gamma isolated from a C57BL/6 brain cDNA library. The mouse and rat PKC-gamma nt and deduced amino acid sequences share 97 and 100% identity, respectively. PMID- 8428670 TI - Synthesis of normal and variant human hypoxanthine-guanine phosphoribosyltransferase in Escherichia coli. AB - Naturally occurring mutations in hypoxanthine-guanine phosphoribosyltransferase (HPRT) have been identified by amino acid sequencing, cDNA cloning, and direct nucleotide sequencing of PCR-amplified transcripts. To determine the effect these mutations have on the catalytic properties of the molecule, knowledge of the three-dimensional structure of HPRT is required. A prerequisite for this, however, is the availability of a large amount of purified product for crystallization and x-ray diffraction analysis. For these reasons we have developed an effective means of producing high levels of human HPRT in Escherichia coli using the expression cassette PCR. By taking advantage of a T7 polymerase/promoter system, we have expressed both normal and variant human hprt sequences in E. coli. The proteins synthesized from these sequences are immunologically and enzymatically active, and are physically indistinguishable from the HPRT in B-lymphoblasts derived from normal and three HPRT-deficient subjects. PMID- 8428671 TI - Organization and sequence of the gene encoding the human acrosin-trypsin inhibitor (HUSI-II). AB - A complete cDNA encoding the acrosin-trypsin inhibitor, HUSI-II, was used as a probe to isolate genomic clones from a human placenta library. Three clones which cover the entire HUSI-II gene were isolated and characterized. The exon-intron organization of the gene was determined and found to be identical to other known Kazal-type inhibitor-encoding genes. The striking similarity in the amino acid sequences which was found previously in HUSI-II and glycoprotein hormone beta subunits, is neither reflected in codon usage nor in the exon-intron arrangement of the genes. A 1.8-kb segment 5' of the gene was sequenced. The analysis of this sequence showed that HUSI-II contains a G + C-rich region upstream from the transcription start point (tsp) which fulfills the criteria for a CpG island. Furthermore, in the first intron, a potential glucocorticoid-responsive element was found as a half-palindrome flanked by two CACCC elements. Determination of the tsp by S1 mapping revealed that HUSI-II has multiple tsp. Genomic Southern hybridization was used to show that HUSI-II is a single-copy gene. The localization of the gene to chromosome 4 was determined by hybridization of a 5' genomic fragment to the DNA of a panel of somatic hybrids between human and rodent cells. PMID- 8428672 TI - Characterization of cDNA clones encoding the human homologue of Saccharomyces cerevisiae ribosomal protein L30. AB - We have isolated cDNA clones encoding the human homologue (hL30) of yeast ribosomal protein (r-protein) L30. The hL30 nucleotide (nt) sequence shows high homology to the yeast sequences and also to a partial Xenopus laevis sequence previously identified as an immunoglobulin heavy chain. The 5' end of hL30 is pyrimidine-rich, as is the case for most other mammalian r-protein mRNAs. The open reading frame consists of 157 codons with a C-terminal region that is different from corresponding regions of the yeast proteins. In several human tissue culture cells, the mRNA encoding hL30 is approx. 700 nt in length. PMID- 8428673 TI - lambda SHK and lambda AASV: phage vectors for efficient cDNA cloning and expression in mammalian cells. PMID- 8428674 TI - Assessment of optic disc topography with scanning laser ophthalmoscope. AB - Evaluation of the topography of the optic disc is of clinical importance to assess the degree of nerve damage. We conducted a study in 17 glaucomatous and 20 control subjects with a scanning laser ophthalmoscope (SLO; Rodenstock) and compared the results with those in conventional photographs. A tomographic image of the cup area in control subjects and the neuroretinal rim area in glaucomatous subjects obtained with the SLO was smaller than that in photographs taken with a conventional camera (P < 0.05 and P < 0.01 respectively). In both the control and glaucoma groups, the optic disc area obtained with the SLO was significantly smaller than that in conventional photographs (P < 0.01). The coefficient of variation of the optic disc parameters studied with the SLO range from 4.2% to 9.1%. A correlation between the optic disc indices studied with SLO and the mean defect of the visual field was statistically significant. The tomographic images obtained by the SLO permit accurate assessment of optic nerve damage. PMID- 8428675 TI - Simultaneous bilateral malignant glaucoma following laser iridotomy. AB - A case of bilateral malignant glaucoma is described in a 50-year-old white woman 4 weeks after successful bilateral laser iridotomy for angle-closure glaucoma. The malignant glaucoma may have been precipitated by systemic hydrochlorothiazide therapy. The condition was worsened by pilocarpine; however, atropine and cyclopentolate opened the angle and deepened the anterior chamber, resulting in resolution of the malignant glaucoma. The clinician must be alert to the possibility of malignant glaucoma following laser iridotomy for angle-closure glaucoma. PMID- 8428676 TI - Heterotransplantation of human uveal melanoma. AB - Cell lines established from the biopsy specimen of a patient with a spindle B choroidal melanoma were transplanted into the posterior choroid of 30 immunosuppressed, pigmented New Zealand rabbits. Growth of the tumor xenografts could be seen 7 to 10 days after transplantation. Tumor xenografts were reproducible and reached an average size of 5 mm in height and 8 mm in their basal dimension by 4 weeks. Histopathology of the original tumor revealed primarily spindle B cells, while the tumor xenografts contained epithelioid, spindle B, and clear cell melanoma cells. Of particular interest was the presence of an extensive and intact vasculature and the absence of a capsule surrounding the lesions in situ. The use of human uveal melanoma cells, the ease of transplantation, and the posterior location of the tumor may make this animal model of use in studies on new diagnostic and therapeutic modalities. PMID- 8428677 TI - Early morphogenesis of persistent hyperplastic tunica vasculosa lentis and primary vitreous (PHTVL/PHPV). Scanning electron microscopic observations. AB - This study provides scanning electron microscopic observations on the early morphogenesis of persistent hyperplastic tunica vasculosa lentis and primary vitreous (PHTVL/PHPV) in canine fetuses at days 28 35 postcoitum (D28 and D35). From previous studies regarding PHTVL/PHPV it is known that a retrolental plaque of fibrovascular tissue is present in eyes of affected canine fetuses from the D33 stage. The contribution of vitreous cells to the formation of the plaque is supported by the results of this study. The lens capsules at the stages described were not found to contain abnormalities such as transparent (thinner) parts or rents, as have been described for postnatal cases of PHTVL/PHPV. These findings support the hypothesis that the capsular anomalies observed in postnatal patients are secondary entities. PMID- 8428678 TI - The effects of intravitreal triamcinolone acetonide on experimental pre-retinal neovascularization. AB - Corticosteroids, alone or in combination with other drugs, have been shown to inhibit angiogenesis. The purpose of this study was to evaluate the efficacy of triamcinolone acetonide in a new model of preretinal neovascularization. Rabbit eyes were treated with intravitreal triamcinolone acetonide 24 h before partial liquefaction of the posterior vitreous with hyaluronidase and injection of 250,000 homologous tissue-cultured dermal fibroblasts. Triamcinolone acetonide effectively inhibited new vessel growth in treated eyes. Only 14% of the treated eyes developed new blood vessels compared to 100% of sham-injected control eyes (P < 0.001). These results suggest that intravitreal triamcinolone acetonide might be effective in inhibiting new vessel growth in patients with inflammatory retinal neovascularization, such as that associated with sarcoidosis or other uveitic syndromes. PMID- 8428679 TI - Tilted discs and central retinal vein occlusion. PMID- 8428680 TI - Acute obstruction of the retinal and choroidal circulation as a complication of interventional angiography. PMID- 8428681 TI - Diffuse unilateral subacute neuroretinitis syndrome in a German most likely caused by the raccoon roundworm, Baylisascaris procyonis. PMID- 8428682 TI - Bilateral cavernous degeneration of the optic nerve associated with multiple arteriosclerotic ischemic infarctions. PMID- 8428683 TI - Immune complex deposition lines in a case of retinal vasculitis. PMID- 8428684 TI - Influence of missed catch trials on the visual field in normal subjects. PMID- 8428685 TI - Morphologic characteristics, proliferation, and tumor marker expression of two human ovarian carcinoma cell lines in three-dimensional culture. AB - Two cell lines derived from human serous ovarian adenocarcinoma (KOC-1S, KOC-2S) were cultured three-dimensionally in type I collagen gel, and their morphology, growth kinetics, and tumor marker expression were compared with those of cells grown on plastic dishes. KOC-1S cells were established from a well-differentiated adenocarcinoma and they formed three-dimensional round colonies and tubular structures in collagen gel culture. KOC-2S cells were established from a poorly differentiated serous adenocarcinoma and they proliferated diffusely in a solid pattern to form irregularly shaped colonies in collagen gel culture. The doubling times of both cell lines were longer in collagen gel culture, but the ratio of their doubling times was virtually the same in both culture systems. KOC-1S cells produced CA125 and TPA, while KOC-2S cells only produced TPA. For both cell lines, tumor marker secretion per 10(5) cells was greater in collagen gel that on plastic dishes. These results suggest that human ovarian carcinoma cell lines retain characteristics similar to those in vivo with regard to histologic type, degree of differentiation, and growth kinetics when they are cultured in collagen gel. PMID- 8428687 TI - Combined antiproliferative activity of 5-fluorouracil and mitomycin-C against primary human ovarian tumors and cell lines in a clonogenic assay. AB - Ovarian tumors from 389 patients were successfully grown in a human tumor clonogenic assay (HTCA). Specimens from patients with or without prior chemotherapy showed similar chemosensitivity patterns. Excluding drugs commonly used for first-line chemotherapy, 5-fluorouracil (5-FU) and mitomycin-C (MMC) are among the active second-line agents, based upon HCTA data. Using this in vitro information from primary tumor specimens, two human ovarian cancer cell lines were used to study in detail the interactions of combined 5-FU and MMC. Drug scheduling which maximized combined antitumor activity was studied. Combined 5-FU and MMC revealed positive drug interactions most consistently when both drugs were used simultaneously with long-term exposure. Pulse treatment with MMC (1 hr) followed by continuous 5-FU exposure resulted in slightly less additive interaction than simultaneous long-term exposure. Pretreatment with 5-FU followed by a continuous exposure to MMC was as effective as the simultaneous method, as long as MMC was added within an 8-hr interval. Drug schedule dependency was examined, revealing that MMC, as a single agent, is both dose and schedule dependent. The results of these in vitro studies suggest that: (1) exposure to prior chemotherapy does not induce demonstrable pleiotropic drug resistance in these ovarian cancers tested; and (2) combined 5-FU and MMC show positive interactions against ovarian cancer cell lines, with optimal scheduling seeming to be long-term simultaneous exposure to both MMC and 5-FU. PMID- 8428686 TI - Development of postmolar trophoblastic disease after partial molar pregnancy. AB - We investigated the clinical characteristics of patients with partial molar pregnancy and patients who developed invasive partial mole. Between 1981 and 1990, 349 patients were followed up by the Aichi prefecture trophoblastic disease registration center after partial molar pregnancy. Ten of the 349 patients with partial molar pregnancy developed invasive partial mole, an incidence (2.87%) significantly lower (P < 0.01) than that for the development of invasive complete mole (174/1410, 12.34%) following complete molar pregnancy during the same period. None of the patients had any histologic evidence of choriocarcinoma after partial molar pregnancy. The medical records of these 10 patients were compared with those of 85 of 174 registered patients with invasive complete mole, whose clinical information could be used. The interval from molar delivery to the diagnosis of invasive mole was under 9 weeks (mean, 4.90 weeks) for the patients with invasive partial mole and was significantly shorter than that for the patients with invasive complete mole, from 2 to 18 weeks (mean, 8.12 weeks) (P < 0.01). All 10 patients with invasive partial mole achieved a negative hCG level (< 0.5 mIU/ml) with eight courses or less of chemotherapy (mean, 5.50 courses). The number of courses was the same as that in the invasive complete mole group (mean, 5.70 courses). None of the patients with invasive partial mole developed recurrence. We conclude that all patients with partial molar pregnancy should be followed up as are those with complete molar pregnancy. Some patients with partial molar pregnancy will develop invasive partial mole with a relatively short interval, but remission can be achieved without the recurrence and choriocarcinoma seldom develops after partial molar pregnancy. PMID- 8428688 TI - Ovarian sex cord tumor with annular tubules: a report of six cases. AB - Six patients suffering from ovarian sex cord tumor with annular tubules (SCTAT) were reported in this article with special reference to the clinical features, histological characteristics, sex hormone profile, and management of disease. SCTAT was documented to be a estrogen-progesterone-secreting tumor based on the observations of glandular atrophy and decidual change of stroma in the endometrium and assays of steroid hormone. Menometrorrhagia followed by persistent amenorrhea and pelvic mass were presented as important clinical features. This tumor was considered as a tumor with low-grade malignancy, and retroperitoneal lymphatic metastasis was thought to be an important pathway of spread. Unilateral salpingo-oophorectomy together with ipsilateral pelvic and para-aortic lymphadenectomy were suggested as an effective treatment for SCTAT. Radiotherapy can be used for local recurrence and distant metastases. PMID- 8428689 TI - Evidence for persistence of mitoxantrone within the peritoneal cavity following intraperitoneal delivery. AB - In clinical trials examining the intraperitoneal (ip) administration of mitoxantrone as therapy of platinum-refractory small-volume residual ovarian cancer, the characteristic "blue color" of the agent has been demonstrated to stain the surface of the peritoneal cavity and to persist for > or = 1 month following the last course of therapy. To determine if this blue staining material contains potentially cytotoxic concentrations of mitoxantrone, we analyzed tissue obtained at exploratory laparotomy in six women who had last received the agent administered ip from 6-22 weeks prior to surgery. Concentrations of mitoxantrone ranged from < 0.1 to 13.8 micrograms/g of tissue examined. Since any mitoxantrone present on the peritoneal surface will be highly protein bound, any residual drug may not have cytotoxic potential. The dose-response curves of mitoxantrone in a human clonogenic cytotoxicity assay against the RPMI 2780/S human ovarian cell line were virtually identical when the cells were incubated in either 5 or 50% fetal bovine serum, suggesting that protein binding will not significantly impair mitoxantrone-induced tumor cell killing. We conclude that the ip administration of mitoxantrone may lead to prolonged exposure of surface tumor to the high local concentrations of the active cytotoxic agent. This effect may contribute significantly to the antineoplastic potential of ip mitoxantrone in patients with small-volume residual ovarian cancer. PMID- 8428690 TI - Recurrent squamous cell carcinoma of the vulva: a study of 73 cases. AB - In a study of 73 patients, diagnosed with recurrent squamous cell carcinoma of the vulva between 1975 and 1990, the effect of clinical variables on the outcome was evaluated. The overall 5-year survival rate was 35.2%. Of the 73 patients, 33 (45.2%) originally had Stage I or II disease and 40 (54.8%) Stage III or IVA; 49 (67.1%) recurred less than 2 years and 24 (32.9%) more than 2 years after initial surgery; and 39 (53.4%) recurred on the vulva only, while 34 (46.6%) recurred beyond the vulva. Of 59 patients who had groin lymph node dissection at initial surgery, 26 (44%) had negative and 33 (56%) had positive nodes. By means of univariate analyses, a significant worsening in outcome was demonstrated with advancing original stage of disease (P < 0.001), positivity of groin lymph nodes (P < 0.01), shortening of recurrence-free interval (P < 0.001), and extension of recurrence beyond the vulva (P < 0.001). In a multivariate analysis (Cox proportional hazards model) recurrence site was the strongest and the only significant predictor of survival. The death risk showed a 3.7-fold increase (95% confidence intervals: 1.6 to 8.7, P = 0.002) for recurrence beyond the vulva over recurrence on the vulva only. For patients who recurred in the vulva only, wide radical local excision provided acceptable survival results, while for all other patients, regardless of type of treatment, the outcome was poor. PMID- 8428691 TI - Studies on ras oncogene activation in endometrial carcinoma. AB - The frequency of K-ras point mutation(PM) at codon 12 was studied in 45 patients with endometrial carcinoma. In vitro amplification of target sequences of DNA extracted from endometrial cancer tissues by polymerase chain reaction and dot blotting with oligonucleotide hybridization were performed. Ten of 45 endometrial carcinomas disclosed K-ras PM at codon 12 (22.2%). Transition from GGT to GAT was most frequent in PM(41.7%). Simultaneously, double PM (GAT/GCT) were also detected in 2 cases. No relationship appeared to be present between PM and clinical prognosis such as clinical stage, histological type, histological grade of differentiation, depth of myometrial invasion, and ascitic cytology. The positive rates of lymph node metastasis tended to be higher in the group with positive PM than in the group without PM. K-ras and C-myc gene amplifications were found in 2 (5.1%) and 3 (7.7%) of 39 cases, respectively. No PM of H-ras at codons 12 and 61 was detected. Our results showed that the PM of K-ras gene at codon 12 was a fairly common event in genetic abnormality and suggested it would have some role in the progression of carcinogenesis in endometrial carcinoma. PMID- 8428692 TI - Characterization of a human ovarian carcinoma cell line: UCI 101. AB - A new epithelial ovarian carcinoma cell line (UCI 101) has been established from the ascitic fluids and solid tumor of a patient with progressive papillary adenocarcinoma of the ovary shown previously to be refractory to combination chemotherapy consisting of cyclophosphamide, Adriamycin, and cisplatin as well as single-agent chemotherapy of taxol and high-dose cisplatin. The UCI 101 cell line grows well with an in vitro doubling time of 24 hr. The cell line expresses the B 72.3 (Tag 72), CA125, MH99 (ESA), and E29 (EMA) cell surface antigens and AE1/AE3 cytokeratins. This cell line overexpresses (as determined by immunocytochemistry) both p-glycoprotein and the epidermal growth factor receptor. The in vitro drug response to single agents including Adriamycin, cisplatin, dequalinium chloride, etoposide, 5-fluorouracil, taxol, and tumor necrosis factor was examined. Intraperitoneal transplantation of the cells into athymic mice resulted in foci of tumor on all peritoneal surfaces including the viscera and diaphragm ultimately leading to solid bulky disease with massive production of ascites. High levels of CA125 (> 500 units/ml) were detected in the serum of tumor-bearing mice. Cytogenetic analysis of cultured cells shows several marker chromosomes containing deletions, duplications, and translocations. Cytologic and histologic evaluation of the xenograft revealed morphological characteristics identical to those of the original tumor. PMID- 8428693 TI - Sensitivity of drug-resistant human ovarian tumor cell lines to combined effects of tumor necrosis factor (TNF-alpha) and doxorubicin: failure of the combination to modulate the MDR phenotype. AB - We have examined four human ovarian tumor lines (A2780, AD10, OVC-8, and SKOV-3) selected for their sensitivity and/or resistance to the recombinant human tumor necrosis factor alpha (TNF-alpha) and the chemotherapeutic drug doxorubicin (DOXO). The tumor lines were either sensitive to both agents, resistant to one or the other, or resistant to both. Of the four lines examined only the DOXO resistant line AD10 exhibited the multidrug-resistance (MDR) phenotype. Enhanced cytotoxicity was seen with the combination of TNF-alpha and DOXO in each line regardless of their sensitivity or resistance patterns and, thus, demonstrates that drug resistance due to the expression of the MDR phenotype or its absence can be overcome by TNF-alpha and DOXO. We then examined whether TNF-alpha or TNF alpha and DOXO modulated the MDR phenotype in AD10 as a possible mechanism of overcoming drug resistance. TNF-alpha had no effect on either DOXO intake or efflux as measured by flow cytometry. Further, TNF-alpha treatment showed no effect on the level of MDR-1 mRNA. These results suggest that the enhanced cytotoxicity seen with the combination of TNF-alpha and DOXO is not the result of any modulation of drug influx or efflux levels by TNF-alpha. Overall, these findings suggest that combination treatment with TNF-alpha and DOXO can overcome resistance inflicted by different mechanisms. PMID- 8428694 TI - Recurrent stage I endometrial adenocarcinoma in the nonirradiated patient: preliminary results of surgical "staging". AB - Recurrent endometrial carcinoma, even when clinically confined to the vagina or pelvis, is associated with poor survival. Pelvic radiotherapy for patients with localized recurrences who have not been previously irradiated has not been highly effective. Our hypothesis was that local salvage therapy fails because a significant number of patients have occult, subclinical distant metastases at the time of relapse. In order to accurately assess disease status at the time of the recurrence, we prospectively evaluated eight patients with recurrent disease limited to the vagina/pelvis by physical examination, routine laboratory tests, and radiologic imaging. All patients underwent a "staging" procedure which included laparotomy, selective pelvic/periaortic lymphadenectomy, peritoneal biopsies, and washings. Three (37.5%) of eight patients had upper abdominal disease found at laparotomy (95% confidence interval 0.11 to 0.71). Presence of subclinical metastases was associated with larger tumor size (> or = 2 cm) and elevated serum CA 125 antigen levels. Treatment was modified in three patients according to the results of surgical staging. One patient was treated with chemotherapy while two patients received whole-abdominal radiation in addition to pelvic fields. Seven of eight patients are alive 21 to 61 months following salvage therapy. Three (43%) of seven patients treated with radiotherapy suffered nonneoplastic bowel obstruction requiring laparotomy at 3, 6, and 15 weeks following completion of radiation therapy. Since 37.5% of patients with recurrent endometrial carcinoma clinically confined to the pelvis had occult upper abdominal disease, surgical reassessment may be warranted, especially in those with elevated serum CA 125 levels or large tumors. Our limited sample size precludes any definitive conclusions regarding our data. Further research will determine the frequency of subclinical metastases and the value of serum CA 125 levels in assessing disease status. PMID- 8428695 TI - Feasibility of intraoperative administration of chemotherapy for gynecologic malignancies: assessment of acute postoperative morbidity. AB - In order to assess the acute effects of the intraoperative administration of intraperitoneal (IP) carboplatin with and without intravenous (IV) doxorubicin in patients with gynecologic malignancies, 25 patients were treated at the conclusion of their surgical procedure. Twenty-three had epithelial ovarian cancer and 2 had advanced endometrial cancer. Twelve patients received IP carboplatin and IV doxorubicin at the conclusion of their primary cytoreduction. The remaining 13 received IP carboplatin alone and consisted of 4 reassessment laparotomies for ovarian cancer, 7 secondary cytoreductions, 1 minilaparotomy for the placement of an IP catheter, and 1 second-look laparoscopy. The median age of the 25 patients was 59 years. Eight patients underwent bowel resections with anastomoses; 2 had ureteral resections and ureteroneocystostomies, 1 required a splenectomy, and 1 underwent a partial hepatic resection. There were no mortalities. Three of the 13 patients who received the IP carboplatin alone had postoperative fevers with no infectious source, which did not delay discharge. No other morbidity was noted in this group. However, 7 patients who received IV doxorubicin along with the IP carboplatin developed severe leukopenia requiring antimicrobial and colony-stimulating factor support. One patient required reexploration for postoperative hemorrhage and also developed a pulmonary embolus. One woman developed postoperative pneumonia. The median hospital stay for the 25 patients was 9 days. Intraoperative chemotherapy can be administered with tolerable immediate adverse effects. PMID- 8428696 TI - DNA ploidy, proliferation index, grade, and stage as prognostic factors for vulvar squamous cell carcinomas. AB - DNA analysis by flow cytometry has been reported to be a useful prognostic technique for a variety of malignant tumors. The goal of this study was to examine DNA ploidy status and proliferation index as potential predictors of recurrence and survival for patients with squamous cell vulvar carcinomas. The DNA ploidy, percentage of cells in the S-phase of the cell cycle, stage, and histologic grade were analyzed in 42 patients. Flow cytometry was performed on archival paraffin-embedded tissue. The histologic specimens were reviewed by a single pathologist. Thirty-four tumors (81%) were diploid and eight (19%) demonstrated an aneuploid peak. The percentage of cells in the S-phase (proliferation index) was calculated for all patients. The 5-year survival rate was 68% for diploid tumors and was 75% for aneuploid tumors. The difference between these rates was not statistically significant (P < or = 0.65). S-phase fraction was also not a useful predictor for recurrence or overall survival. FIGO stage and histologic grade were accurate predictors for both recurrence and 5 year survival. Multiple predictor analysis using stage, grade, ploidy status, and proliferation index did not identify any subgroup which would predict recurrence better than FIGO stage and grade. DNA ploidy and S-phase fraction analysis do not appear to be clinically useful prognostic factors for vulvar squamous cell carcinomas. PMID- 8428697 TI - The validity and significance of substages of advanced ovarian cancer. AB - No studies to date have evaluated the validity of the new FIGO substaging of advanced epithelial ovarian cancer nor assessed the importance of substage in relation to other elements such as age at diagnosis, debulking surgery, and second-look laparotomy. The purpose of this study was to determine the significance of these factors. One hundred sixty-seven patients with Stage III ovarian cancer were restaged according to the 1988 FIGO criteria (6% Stage IIIa, 15.6% Stage IIIb, and 78.4% Stage IIIc). The mean age at diagnosis was 40.5 for Stage IIIa, 51 for Stage IIIb, and 62 for Stage IIIc (P = 0.0001). Median survival was 2.5 years for patients age < 60 and 1.4 years for those age > or = 60 (P = 0.0001). Median survival for patients undergoing TAH/BSO was 2.06 years, bowel resection 1.39 years, and biopsy only 1.38 years (P = 0.0003). Only 61 of 131 Stage IIIc patients underwent second-look laparotomy. Seven of nine Stage IIIa, 6 of 17 Stage IIIb, and 14 of 61 Stage IIIc patients had negative second look laparotomies (P = 0.004). Only 4 of the 14 patients with Stage IIIc and 8 of 13 Stage IIIa/b patients are alive after negative second look (P = 0.37). Median survival for Stage IIIa patients has not been reached and for Stages IIIb and IIIc was 2.29 years and 1.33 years, respectively (P = 0.0001). These data confirm the prognostic validity of FIGO substages for Stage III. The age differential by substages suggests that the natural history of Stage III disease is progressive over several decades. The appropriateness of aggressive cytoreductive surgery and second-look laparotomy must be reevaluated using the new FIGO staging system. PMID- 8428698 TI - Prognostic significance of DNA content and nuclear morphology in borderline ovarian tumors. AB - We used the technique of image analysis to simultaneously measure DNA content and nuclear morphology of 21 borderline ovarian tumors. Aneuploidy was identified in 9 of 21 tumors and was unrelated to tumor stage or nuclear grade. Morphometric nuclear features that were measured included size, shape, texture, and average density. Nuclear size and shape were positively correlated (r = 0.507), and nuclear size and average density were negatively correlated (r = -0.772). Six tumors recurred and recurrence was significantly associated with tumor aneuploidy (P = 0.046), stage III tumors (P = 0.03), and increased nuclear texture (P = 0.07). These results suggest that measurement of DNA ploidy and nuclear morphology using image analysis can provide important prognostic information in patients with borderline ovarian tumors. PMID- 8428700 TI - Metastatic clear-cell hidradenocarcinoma of the vulva. AB - Clear-cell hidradenocarcinoma is a malignant tumor of sweat gland origin. It is most often found on the trunk, head, and extremities. This case report describes a rare occurrence of this tumor on the vulva of a young woman. The discovery of metastatic disease reflects the potentially aggressive nature of this tumor. PMID- 8428699 TI - Effectiveness of two barriers at inhibiting post-radical pelvic surgery adhesions. AB - OBJECTIVE: To compare the abilities of Gore-Tex Surgical Membrane (Gore-SM) and Interceed to inhibit post-radical pelvic surgery adhesion formation in a porcine model of radical hysterectomy, salpingo-oophorectomy, and resection of pelvic peritoneum. METHODOLOGY: Twenty-seven hogs were evaluated. After completion of the radical resection animals were randomized to no treatment or covering the peritoneal defect with a tailored sheet of Gore-SM or Interceed. The Gore-SM was secured in place using a continuous running 4-O Prolene suture while the Interceed was allowed to adhere without sutured attachment. Four weeks after the initial surgery, the animals were reexplored, adhesions quantified, and note was made of any segments of small bowel that were adherent into the pelvis. RESULTS: One animal in each of the treatment groups died of respiratory compromise in the postoperational period. Adhesion scores for the Interceed covered pelvis (n = 9; mean = 0.04 +/- 0.04) were significantly less than those treated with Gore-Sm (n = 9, mean = 0.18 +/- 0.15, P = 0.02). Both treated groups had significantly fewer adhesions in the pelvis than did the controls (n = 7, mean 1.33 +/- .51; vs Interceed P = 0.0005, vs Gore SM, P = 0.0007). CONCLUSIONS: Interceed was a more effective barrier for postoperative adhesion prophylaxis than the Gore-SM. Interceed also offered a technical advantage at time of placement. PMID- 8428701 TI - Spinal cord compression in metastatic cervical cancer. AB - Spinal cord compression secondary to metastatic cervical cancer may not be considered as a possible cause of neurologic symptoms by primary care physicians who do not often treat these patients. Delays in diagnosis and treatment may result in irreversible but potentially preventable neurologic changes. This report describes 5 cases of spinal cord compression in patients with metastatic cervical carcinoma, 2 of whom were previously undiagnosed with cervical cancer. These 2 patients represent 1.6% (2 of 121) of all new cervical cancer cases diagnosed during this time period. Two of 5 patients (40%) with spinal cord compression showed improvement following therapy by regaining the ability to walk, while none of the remaining patients had further acute deterioration of neurologic function. The mean survival of patients presenting with spinal cord compression from cervical cancer in this series was 4 months (maximum 6 months). This series illustrates the relative frequency with which spinal cord compression is seen in patients with a new diagnosis of invasive cancer. This diagnosis should be considered when evaluating neurologic complaints in known cervical cancer patients or any woman with apparent pelvic pathology. Rapid diagnosis and treatment of these lesions, while not likely to improve overall survival significantly, can improve function and alleviate symptoms. PMID- 8428702 TI - Uterine mixed embryonal rhabdomyosarcoma and fetal rhabdomyoma. AB - A polypoid uterine tumor, occurring in a 31-year-old woman, with histopathologic features of both embryonal rhabdomyosarcoma and fetal rhabdomyoma is reported. The clinical and pathologic differences between pure fetal rhabdomyoma and embryonal rhabdomyosarcoma are detailed and theories concerning the histogenesis of such a mixed, or intermediate, tumor are discussed. Most likely, this neoplasm represents one example in a spectrum of tumors with varying proportions of immature and mature skeletal muscle elements. Its good prognosis is in keeping with that of uterine (cervical) embryonal rhabdomyosarcomas in general, although the presence of mature skeletal muscle elements may signify an even better prognostic group of tumors. PMID- 8428703 TI - Myxoid leiomyosarcoma of the uterus. AB - BACKGROUND: A case of myxoid leiomyosarcoma of the uterus is presented. In contrast to 10 of the 11 previously published cases, our patient has a tumor with a high mitotic rate. CASE: The patient presented with a 2-month history of vaginal bleeding and a 20-week-size uterine mass. A total abdominal hysterectomy with bilateral salpingo-oophorectomy was performed. At the time of surgery, the tumor appeared well circumscribed. Microscopically, an infiltrative growth pattern, bizarre nuclei, and 30 abnormal mitotic figures per 10 high power fields were noted. The patient was treated with ifosfamide and mesna. After the fifth course of chemotherapy, a pelvic mass was palpated. Exploratory laparotomy demonstrated a large sidewall recurrence and upper abdominal metastases which were completely resected. The patient is currently being treated with combination chemotherapy consisting of vincristine, Adriamycin, and cyclophosphamide. CONCLUSION: Ten of the 11 previously reported cases of myxoid leiomyosarcoma had a low mitotic count. However, our patient has a high mitotic count in combination with an infiltrative growth pattern. Ifosfamide was ineffective as adjuvant chemotherapy and there was aggressive tumor growth while the patient was on this regimen. PMID- 8428704 TI - Tissue-specific alterations in lipoprotein lipase activity in the rat after chronic infusion of isoproterenol. AB - This study describes the consequences of sustained exposure to the nonspecific beta adrenergic agonist isoproterenol on the activity of lipoprotein lipase (LPL) in tissues that are sites for a large portion of the intravascular hydrolysis of triacylglycerols. Chronic exposure to isoproterenol was accomplished by means of osmotic pump infusion during four consecutive days, and assessment of the activity of LPL was performed at the end of the treatment. Four days of ISO infusion resulted in a large decrease in LPL activity of parametrial white adipose tissue, which declined to half of that found in the untreated group. In contrast, brown adipose tissue LPL activity was greatly increased (approximately threefold) by ISO treatment. The beta agonist raised LPL activity in red vastus lateralis muscle by a factor of four over control values, whereas heart enzyme activity was not significantly altered by the drug. These results demonstrate that catecholamines are part of the factors which determine the tissue specificity of LPL regulation. The tissue-specific changes in enzyme activity in response to isoproterenol, which are in very good agreement with the known metabolic actions of catecholamines in adipose and skeletal muscle tissues, attest to the key role of the enzyme in the partitioning of lipid substrates in situations of increased sympathetic activity. PMID- 8428705 TI - Differences between plasma viscosity and proteins of types 1 and 2 diabetic Africans in early phase of diabetes. AB - Plasma and serum viscosity and associated protein titres were determined in short and long term types 1 and 2 diabetic Africans. The viscosity of plasma and serum, fibrinogen, total serum protein and albumin concentrations of both short- and longterm type 1 diabetics were significantly greater than those of the nondiabetic controls, although no significant differences in these parameters were recorded between the two groups of type 1 diabetics. There were no significant differences between short term type 2 diabetics and the controls in the parameters examined except albumin which is significantly higher in the former. However, plasma and serum viscosity, fibrinogen, total serum protein protein and albumin of longterm type 2 diabetics were significantly elevated above those of the control group. These results show that there is an earlier presentation of cardiovascular risk factors in type 1 than in type 2 diabetic Africans. PMID- 8428707 TI - Effect of hypermagnesemia on the adenohypophyseal-gonadal function, parathyroid hormone secretion and some other hormonal indicators. AB - Hormonal secretory mechanisms are influenced positively or negatively by magnesium, the effect depending on its concentration. The submitted study is concerned with the effect of acute hypermagnesemia induced by intravenous infusion of 6 g MgSO4 on quiescent and LHRH-induced secretion of FSH, LH, prolactin and testosterone; on the levels of intact parathyroid hormone; and on other hormonal and mineral indicators in 10 healthy male subjects. Gonadotropin secretion was not altered by hypermagnesemia. Quiescent testosterone levels decreased moderately (p < 0.01), and the secretory kinetics of the hormone slightly changed. Parathyroid hormone levels markedly dropped in the 60th minute of hypermagnesemia duration (p < 0.01), the drop depending on the initial values of the hormone prior to the administration of magnesium (r = 0.923, p < 0.01). Although in hypermagnesemia no correlations between the levels of parathyroid hormone and those of magnesium, calcium and phosphorus could be found, in normomagnesemia significant correlations were confirmed between the levels of parathyroid hormone and magnesemia (r = -0.8068, p < 0.05), and the levels of parathyroid hormone and calcemia (r = -0.9451, p < 0.01). In hypermagnesemia there were no alterations in the levels of the plasma prolactin, cortisol or catecholamines, nor was there any alteration in the rate at which catecholamines were excreted in urine. In conclusion, magnesium in supraphysiologic concentrations does not significantly change the function of the adenohypophyseal gonadal axis or other hormonal indicators studied. The slight decline of quiescent testosterone levels does not seem to have any clinical significance, although this must be verified by further studies.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428706 TI - Effects of nitrendipine on blood pressure, renin-angiotensin-system and atrial natriuretic peptide in hypertensive type I diabetic patients. AB - In this study, the effects of a six week treatment with the calcium channel blocking agent nitrendipine were assessed in 20 hypertensive type I diabetic patients who received a single oral dose of 20 mg daily. Plasma concentrations of atrial natriuretic peptide (ANP), plasma renin activity (PRA) and active renin, aldosterone, glycosylated hemoglobin (HbA1) and fructosamines were determined at the beginning and the end of the study. Interactions of these endocrine parameters with blood pressure behaviour were analyzed by a correlation matrix. In response to the drug treatment, the blood pressure was significantly lowered from 158.0/97.2 to 145.7/88.2 mmHg. The plasma concentrations of ANP were also significantly reduced from 106.8 to 89.7 pg/ml. There were no changes in serum aldosterone, while PRA and active renin exhibited a significant increase following the six week treatment. HbA1 and fructosamines remained unaltered. There were no significant correlations for ANP and blood pressure values, as well as for ANP and PRA or aldosterone. We did find, however, a significant correlation of the ANP values with the difference of the systolic blood pressure levels pre- and post-treatment. These data fully confirm the blood pressure lowering properties of this calcium channel blocker and its possible interference with steroidogenesis, since the effects of increased PRA on aldosterone secretion were clearly blunted. The lowering of plasma ANP levels may represent a decreased ANP secretion due to calcium channel blockade or might also be due to the natriuretic effects of nitrendipine, thus allowing ANP levels to decline as a function of lessened sodium retention.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428708 TI - Role of dehydroepiandrosterone and dehydroepiandrosterone sulfate for the maintenance of axillary hair in women. AB - To delineate the relationship between sex hormones and the axillary hair in women, serum concentrations of dehydroepiandrosterone, dehydroepiandrosterone sulfate and testosterone were measured in 177 normal women (aged 16 to 76 y). Both menstrual cycle and axillary hair were present in all normal women younger than 50 years. In the 6th decade, axillary hair was present in 21 of 42 normal women but menstruation was present only in 8 of the 42 women. In the 7th and 8th decades, axillary hair was present in 36% and 25% of them, respectively. None of the subjects over 60 years of age had a menstrual cycle. Serum dehydroepiandrosterone and dehydroepiandrosterone-sulfate concentrations were significantly higher in axillary hair-positive than in -negative women after 50 years of age. Serum testosterone concentration was low in normal women after 50 years of age compared to those of under 49 years, and it was marginally lower in axillary hair-negative than in -positive normal women. However the difference between the axillary hair-positive and -negative subjects was not statistically significant. It is suggested that weak adrenal androgens such as dehydroepiandrosterone and dehydroepiandrosterone-sulfate, rather than testosterone, play an important role for the maintenance of axillary hair in aged women. PMID- 8428709 TI - Lack of effect of octreotide on parathyroid hormone levels and urinary excretion of epidermal growth factor. AB - To determine whether octreotide may acutely modify calcium homeostasis and/or inhibit the EGF production by the kidney, a subcutaneous injection of octreotide (100 micrograms) was administered to ten healthy volunteers. Blood and urine samples were collected at -30, 0, 60, 90, 120 and 180 minutes. Serum insulin, intact PTH, osteocalcin, total and ionized calcium and urinary EGF and creatinine were measured. There was no modification in serum PTH, osteocalcin and total or ionized calcium. There was no significant modification of urinary EGF, although a trend to decrease was evidenced. PMID- 8428710 TI - Fluctuations of carbohydrates in haemolymph of honeybee (Apis mellifica) after fasting, feeding and stress. AB - The haemolymph of honeybee contains trehalose, a non-reducing disaccharide, glucose and fructose. The aim of this work was to describe changes of haemolymph sugar induced by different physiological situations. Gas chromatography for the determination of carbohydrate concentrations in the haemolymph was introduced. The individual bee was anesthetized by CO2 and punctured at tergum III in order to gain 1 microliter haemolymph using a glass capilette. The main sugars in the haemolymph were determined as silylated derivatives by gas chromatography. Bees living under exactly standardized conditions in an artificial bee-house were observed after feeding and fasting. In addition saccharides were determined after "run-stress" along different distances. PMID- 8428711 TI - Levels of parathyroid hormone-related protein (PTHrP) in hypercalcemia of malignancy are not lowered by treatment with the bisphosphonate BM 21.0955. AB - Parathyroid hormone-related protein (PTHrP) is a major cause of hypercalcemia in malignancy and serum levels are elevated in many patients suffering from this syndrome. In 10 patients with hypercalcemia of malignancy the levels of the midregional fragment of PTHrP in serum were determined by radioimmunoassay over 7 days during a calcium-lowering treatment with a single dose of the bisphosphonate BM 21.0955. PTHrP concentrations remained unchanged 6 days after administration of the drug as compared with pretherapeutic values, thus apparently excluding an effect of either the drug itself or the rapid fall in serum calcium on the release of PTHrP by the tumors or on its clearance from the circulation. In the patients with elevated midregional PTHrP levels (n = 6), the calcium-lowering effect of the drug was significantly less pronounced than in patients with normal PTHrP (n = 4) (mean serum calcium of 2.89 vs. 2.51 mmol/l at day 6), despite similar pretherapeutic concentrations. Of the six patients with elevated PTHrP, five were still hypercalcemic, whereas in the group with normal PTHrP one out of four patients remained hypercalcemic. In conclusion, PTHrP levels in hypercalcemia of malignancy remained unchanged after calcium-lowering therapy with bisphosphonates. High serum PTHrP levels were, however, predictive of a lesser effectiveness of the drug in lowering serum calcium. PMID- 8428712 TI - Peripheral muscle glucose and potassium transport in acromegaly. PMID- 8428713 TI - Effect of fatty acid oxidation inhibition on glucose metabolism in diabetic rats. AB - Inhibition of fatty acid (FA) oxidation has been shown previously to lower blood glucose levels acutely in diabetic rats. However, the longer term effects of FA oxidation inhibition have not been determined. This study examines the effect of inhibition of (FA) oxidation for 3 weeks on carbohydrate metabolism in rats rendered diabetic with streptozotocin (STZ). STZ treated rats (50 mg/kg) were randomized into 3 groups: a non-treated diabetic group and 2 groups treated with either 12.5 or 25 mg/kg/day Etomoxir (a specific carnitine palmitoyl transferase inhibitor) for 3 weeks. The 3 groups of rats had ad libitum access to a high fat diet (50% energy fat, 30% carbohydrate and 20% protein) throughout the study. After 3 weeks hepatic glucose production (HGP) was estimated using a constant infusion of (3H)-6-glucose in-vivo after an overnight fast. Inhibition of FA oxidation in diabetic rats resulted in a significant reduction in fasting glucose levels and hepatic glucose production. In addition, experiments with adipocytes isolated from diabetic rats treated with etomoxir demonstrated an increase in sensitivity and responsiveness to insulin of glucose utilization and pyruvate dehydrogenase (PDH) activity. It is important to note that these improvements in carbohydrate metabolism were not accompanied by increases in circulating FFA or triglyceride levels which were unchanged or lower after inhibition of FA oxidation. PMID- 8428714 TI - Persistence of systemic and splanchnic hyperkinetic circulation in liver transplant patients. AB - Portal pressure and portal-systemic collateral circulation after orthotopic liver transplantation have not been investigated. We studied systemic and splanchnic hemodynamics in 17 patients with cirrhosis before and 205 +/- 146 days after orthotopic liver transplantation. Among the 17 orthotopic liver transplantation patients, 12 had undergone hemodynamic study in the 6 mo before orthotopic liver transplantation. Controls were 50 patients with normal liver architecture. Cardiac index remained elevated in orthotopic liver transplantation patients compared with controls (3.6 +/- 0.9 vs 3.1 +/- 0.4 L/min.m2; p < 0.05). Azygos blood flow, which was elevated before orthotopic liver transplantation (585 +/- 402 ml/min), remained elevated after orthotopic liver transplantation (553 +/- 343 ml/min). In transplant patients, hepatic blood flow was higher than it was in controls (2.26 +/- 0.86 vs. 1.38 +/- 0.57 L/min; p < 0.05). Elevated hepatic blood flow correlated with cardiac index (r = 0.647, p < 0.025). In patients with normal graft function, hepatic venous pressure gradient was normal (2 +/- 1 mm Hg). In a patient with acute rejection, a sharp elevation in the hepatic venous pressure gradient was observed; it returned to normal 45 days after treatment. We conclude that despite normal portal pressure, portal-systemic collateral blood flow remains elevated after orthotopic liver transplantation. Possibly because of persistent collateral circulation, which may keep portal tributary blood flow elevated, hepatic blood flow is increased after orthotopic liver transplantation. Elevated splanchnic blood flow, in turn, contributes to the high cardiac index in liver recipients. Finally, a sharp elevation in portal pressure, which may be observed during acute rejection and subsides after treatment, merits further study. PMID- 8428715 TI - Prospective controlled trial of selective parenteral and enteral antimicrobial regimen in fulminant liver failure. AB - To compare the efficacy of a selective parenteral and enteral antimicrobial regimen in patients with fulminant liver failure, we classified 104 patients on reaching grade II encephalopathy as infected or non-infected. Patients who were infected were randomly assigned to receive IV cefuroxime (group 1) or selective parenteral and enteral antimicrobial regimen (group 2). Noninfected patients were randomly selected to receive either selective parenteral and enteral antimicrobial regimen (group 3) or no initial antimicrobials until clinically indicated (group 4). The four groups were comparable regarding age, sex, cause of disease, coma grade, international normalization ratio, presence of kidney failure and indicators of poor prognosis on admission to the study. Clinical parameters such as white cell count, temperature or changes in the chest radiograph, which were used to stratify patients into those infected or not, were not good predictors of infection because early infection rates were similar in the two groups. Three patients died within 24 hr and were excluded from the analysis. We found 42 microbiologically confirmed infections: group 1, 6 of 21; group 2, 8 of 21; group 3, 9 of 28; and group 4, 19 of 31. A reduction in infection was seen between groups 3 and 4 (p < 0.05). Patients receiving the selective parenteral and enteral antimicrobial regimen (groups 2 and 3) had fewer infections than the control group (group 4) (p < 0.005). Groups receiving early antimicrobial therapy (groups 1, 2 and 3) had a lower incidence of infection compared with group 4 (p < 0.0005). Overall, 55.5% survived, with no significant difference between the four groups.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428716 TI - Continuous increase of alcohol dehydrogenase activity along the liver plate in normal and cirrhotic human livers. AB - To determine the zonal distribution of alcohol dehydrogenase in normal and cirrhotic human livers, we measured activities of this enzyme by quantitative cytochemical analysis along the liver cell plate in liver specimens from 10 normal organ donors and from 7 children with extrahepatic biliary atresia cirrhosis. In normal human liver samples, a continuous increase in alcohol dehydrogenase activity was observed along the sinusoid from the periportal to the perivenular hepatocytes (mean extinction units from 16.2 +/- 10.0 to 58.0 +/- 14.8). A similar observation was made in cirrhotic nodules, with activity increasing continuously from nodule periphery to center (7.6 +/- 4.1 to 44.9 +/- 13.3). This study demonstrates a heterogeneous pattern of alcohol dehydrogenase distribution along the sinusoid in normal human liver specimens. In addition, demonstration of this heterogeneity in human cirrhosis suggests that the cirrhotic liver is able to maintain a parenchymal functional organization, with persistence of metabolic zonation. PMID- 8428717 TI - Eosinophilic infiltration of the liver in primary biliary cirrhosis: a morphological study. AB - Eosinophils and their secretory proteins are important in necroinflammatory processes. In this study we surveyed eosinophilic infiltration of the liver in 176 liver biopsy specimens from patients with primary biliary cirrhosis. A considerable number of eosinophils were found in the portal tracts in 51 of these specimens (29%), whereas this finding occurred in only 2 of 50 (4%) specimens from patients with viral chronic active hepatitis. Eosinophilic infiltration of the portal tracts was focal. These eosinophils were immunohistochemically shown to contain the secreted form of eosinophilic cationic protein. Grades of eosinophilic infiltration of the portal tracts were higher in those livers with higher grades of lymphoplasmacytic infiltration, granulomas and florid duct lesion in the portal tracts, and they were lower in those livers with higher grades of bile duct disappearance and orcein-positive granuoles in hepatocytes. In the livers of patients with primary biliary cirrhosis, eosinophils were abundant around the damaged bile ducts, around the portal vein branches and at the marginal regions, and they were scattered in the portal tracts, whereas only the last pattern was seen in viral chronic active hepatitis. Peripheral blood eosinophils tended to be increased in primary biliary cirrhosis patients with higher grades of eosinophilic infiltration of the portal tracts. Eosinophilic infiltration of the portal tracts may play a role in immunological injuries of the interlobular bile ducts in primary biliary cirrhosis. PMID- 8428718 TI - Effects of propranolol on gastric mucosal perfusion in cirrhotic patients with portal hypertensive gastropathy. AB - This study investigated the effects of the short-term administration of propranolol on gastric blood perfusion in cirrhotic patients with portal hypertensive gastropathy. Portal hypertensive gastropathy is a common cause of nonvariceal bleeding in cirrhosis, which is associated with increased gastric mucosal perfusion and is favorably influenced by propranolol therapy. Gastric mucosal perfusion was evaluated with laser-Doppler flowmetry and reflectance spectrophotometry. Measurements were performed under basal conditions and after the double-blind administration of propranolol (0.15 mg/kg intravenously) or placebo. Propranolol administration significantly reduced (p < 0.001) the laser Doppler signal (2.93 +/- 0.23 vs. 2.25 +/- 0.22 V) and the hemoglobin content of the gastric mucosa (99.2 +/- 3.8 vs. 89.3 +/- 3.1 arbitrary units), whereas the oxygen content remained unchanged (37.4 +/- 1.2 vs. 36.9 +/- 1.0 arbitrary units). Placebo administration had no effect in any of these parameters. Changes in gastric perfusion after propranolol administration were associated with a significant decrease in hepatic venous pressure gradient and azygos blood flow. We conclude that short-term propranolol administration, in addition to lowering portal pressure, reduces the increased gastric blood perfusion in cirrhotic patients with portal hypertensive gastropathy, an effect that may contribute to prevention of bleeding from these lesions. PMID- 8428719 TI - Renal vasoconstriction in cirrhosis evaluated by duplex Doppler ultrasonography. AB - Studies of renal perfusion when kidney function tests are still normal could be useful to understand the pathophysiology of functional kidney impairment in cirrhosis; currently, this requires invasive methodology. Duplex Doppler ultrasonography allows noninvasive evaluation of intrarenal arterial resistances. In 19 nonascitic and 35 ascitic cirrhotic patients with normal kidney function (normal serum creatinine and urea levels) and in 17 controls, we measured the intrarenal arterial pulsatility index (Pulsatility index = [Peak systolic velocity-Minimum diastolic velocity]/Mean velocity) and the resistive index (Resistive index = [Peak systolic velocity-Minimum diastolic velocity]/Peak systolic velocity) by duplex Doppler ultrasonography after visualization of interlobar, interlobular or arcuate arteries by color Doppler ultrasonography. The pulsatility index and resistive index (calculated as the mean of three to five consecutive determinations) were significantly higher in cirrhotic patients than in control patients (pulsatility index: 1.16 +/- 0.24 vs. 0.78 +/- 0.05 [mean +/- S.D.], p < 0.001; resistive index: 0.67 +/- 0.06 vs. 0.53 +/- 0.03, p < 0.001). The pulsatility index and resistive index were significantly higher in nonascitic cirrhotic patients than in control patients, in ascitic patients than in nonascitic patients, in ascitic patients not treated with diuretics than in nonascitic ones and in ascitic patients treated with diuretics than in those not treated. The pulsatility index and resistive index measured before and after treatment with diuretics (7 to 15 days) in seven patients were significantly increased by treatment. The pulsatility index and resistive index were significantly higher in Child-Turcotte-Pugh class B and C patients than in class A patients.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428721 TI - Portal hypertension in lymphoproliferative and myeloproliferative disorders: hemodynamic and histological correlations. AB - The pathogenesis of portal hypertension in patients with lymphoproliferative and myeloproliferative disorders is not fully understood. We investigated 20 patients with myeloproliferative disease and 47 patients with lymphoproliferative disease. Transvenous liver biopsies and hepatic vein pressure gradient measurements were performed in all patients, and portal vein blood flow was measured by pulsed Doppler sonography in 31 of these patients and in 22 normal volunteers. The hepatic vein pressure gradient was significantly higher in patients with hepatic infiltrates, fibrosis or both than in patients without hepatic lesions (8.3 +/- 5.0 mmHg vs. 4.1 +/- 2.3 mmHg; p < 0.01). Portal vein blood flow was significantly higher in patients with hematological disease than in normal volunteers (31.2 +/- 15.5 ml/min.kg vs. 14.2 +/- 4.6 ml/min.kg;p < 0.01). In 81.8% of patients with hepatic infiltrates, fibrosis or both and increased portal vein blood flow, the hepatic vein pressure gradient was greater than 6 mm Hg. Although we saw a significant correlation between splenic vein blood flow and portal vein blood flow (n = 20; p < 0.01), we found no significant correlation between splenic vein blood flow and hepatic vein pressure gradient or spleen size. Hepatic infiltration and fibrosis appear to be major determinants of increased hepatic vein pressure gradient, probably because they increase intrahepatic vascular resistance. The role of increased splenic blood flow is probably not determinant. However, because portal pressure was not measured directly in this study, the incidence of portal hypertension may have been underestimated. PMID- 8428720 TI - Induction of cytochrome P-4502E1 in the human liver by ethanol is caused by a corresponding increase in encoding messenger RNA. AB - The propensity of centrilobular liver damage to develop in alcohol abusers after exposure to various hepatotoxins, including ethanol itself, has been linked to the induction by ethanol of P-4502E1, a microsomal P-450 enzyme that bioactivates these agents to reactive metabolites. Whereas long-term ethanol consumption elicits a marked increase in hepatic P-4502E1 content, the molecular mechanism by which ethanol produces this effect is the subject of controversy in animals, and it has not been elucidated in human beings. Possible mechanisms include increased enzyme synthesis stemming from elevated 2E1 messenger RNA levels, enhanced translation of preexisting messenger RNA or stabilization of P-4502E1 protein. To determine which, if any, of these mechanisms underlies P-4502E1 induction in human beings, we examined the effects of ethanol intake on the hepatic intralobular distribution of P-4502E1 messenger RNA and the corresponding protein. Liver sections derived from needle biopsy specimens were obtained from five recently drinking alcoholics (last drink no more than 36 hr before) and eight control subjects (five abstaining alcoholics [last drink 96 hr or more before] and three nondrinkers). In situ hybridization of these liver sections with a human P-4502E1 complementary DNA probe was used to localize P-4502E1 messenger RNA transcripts. Quantitative image analysis of hybridized sections from control subjects revealed that P-4502E1 transcript content in perivenular (zone 3) hepatocytes was significantly higher (p < 0.05) than in midzonal (zone 2) and periportal (zone 1) cells (18.3 +/- 1, 9.5 +/- 2 and 3.1 +/- 2 arbitrary density units, respectively; mean +/- S.E.M.). In recent drinkers, acinar regions containing P-4502E1 transcripts were elevated 2.9-fold compared with those in controls (32.8% +/- 7% vs. 11.2% +/- 2%; p < 0.01), with this messenger RNA increase occurring mainly in perivenular cells (29.6 +/- 3 vs. 18.3 +/- 1 units; p < 0.01). P-4502E1 protein distribution, assessed by the immunohistochemical staining of liver sections with P-4502E1 antibodies, was found to be analogous to that of the messenger RNA in control subjects (the level in perivenular cells was greater than that in midzonal cells, which was greater than that in periportal cells), whereas recent drinkers exhibited marked elevations in enzyme content in both perivenular and midzonal hepatocytes. Moreover, cellular levels of P-4502E1 protein and messenger RNA were significantly correlated (rs = 0.79; p < 0.001) in all patients.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8428722 TI - Spontaneous bacterial peritonitis in cirrhosis: predictive factors of infection resolution and survival in patients treated with cefotaxime. AB - Cefotaxime is the most commonly used antibiotic for initial therapy of spontaneous bacterial peritonitis in cirrhosis. However, since the introduction of cefotaxime no study has been performed to investigate factors influencing prognosis in cirrhotic patients with this type of infection. In this study, predictive factors for infection resolution and patient survival were investigated in 213 consecutive episodes of spontaneous bacterial peritonitis in 185 cirrhotic patients. All patients were initially treated with cefotaxime. One hundred sixty-five episodes (77%) resolved with cefotaxime alone, and two more episodes (1%), initially unresponsive to cefotaxime, were cured after modification of antibiotic therapy. In a multivariate analysis (stepwise logistic regression), only 4 of 51 clinical and laboratory variables obtained at the time of diagnosis of infection were identified as independent predictors of infection resolution: band neutrophils in white blood cell count, community-acquired vs. hospital-acquired peritonitis, blood urea nitrogen level and serum aspartate aminotransferase level. No patient experienced serious adverse effects related to cefotaxime. Eighty-two patients died during hospitalization (38% mortality rate in relation to the 213 episodes of peritonitis). In the multivariate analysis, six variables were independently correlated with survival: blood urea nitrogen level, serum aspartate aminotransferase level, community-acquired vs. hospital acquired peritonitis, age, Child-Pugh score and ileus. No microbiological data had predictive value for infection resolution or survival.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428723 TI - Development of a bioartificial liver: properties and function of a hollow-fiber module inoculated with liver cells. AB - We have developed a bioartificial liver support system utilizing hollow-fiber bioreactor, plasmapheresis and microcarrier cell culture technologies. Liver cells were obtained through portal vein perfusion with ethylenediaminetetraacetate or ethylenediaminetetraacetate/collagenase. A mathematical model of mass transport in a hollow-fiber module, at various plasma flow velocities and system configurations, was developed. The bioartificial liver's ability to carry out specific differentiated metabolic liver functions was tested in vitro and in vivo. A reproducible large-animal model of acute ischemic liver failure was developed. Most major first-generation cyclosporine and 19-norterstosterone metabolites were isolated after substrate addition to the bioartificial liver in vitro. After bioartificial liver treatment for 6 hr (with dog or pig liver cells), dogs with acute liver failure had significantly lower serum ammonia and lactate levels and significantly higher serum glucose levels than did control animals treated with a bioartificial liver system inoculated with microcarriers alone. In addition, bioartificial liver-treated animals had significantly higher mean systolic blood pressures than did controls. Liver cell viability at the end of the 6-hr in vivo experiment was greater than 90%. PMID- 8428724 TI - Hyaluronic acid uptake by the isolated, perfused rat liver: an index of hepatic sinusoidal endothelial cell function. AB - Previous studies indicate that sinusoidal endothelial cells bind and internalize hyaluronic acid at much greater rates than do other liver cells. Thus hepatic hyaluronic acid removal rate may be indicative of sinusoidal endothelial cell function. In these studies the uptake of hyaluronic acid (molecular weight 1.3 x 10(6)) was measured in isolated perfused rat liver under a variety of conditions. Uptake was dependent on hyaluronic acid concentration. At all concentrations tested, the rate of hyaluronic acid uptake stabilized at a steady-state level 2 to 3 min after development of a high rate of apparent uptake. At saturating hyaluronic acid concentration (150 ng.ml-1), the steady-state uptake rate was 10.4 +/- 1.0 micrograms.gm-1 liver wet wt.hr-1, which is as high as or higher than the rates reported for isolated rat liver sinusoidal endothelial cells. The half-maximal rate of uptake was attained at a hyaluronic acid concentration of 80 ng.ml-1. Hyaluronic acid uptake was inhibited by heparin (80%), a competitive ligand for the hyaluronic acid receptor on sinusoidal endothelial cells; 4 beta phorbol 12 beta-O-myristoyl 13 alpha-acetate (25% to 50%), a tumor promoter and activator of protein kinase C; prostaglandin F2 alpha (24% to 52%), an eicosanoid secreted in the liver by Kupffer cells; A23187 (33% to 66%), a Ca2+ ionophore; and Escherichia coli lipopolysaccharide (16% to 43%). Platelet activating factor did not affect hyaluronic acid uptake by the perfused liver. Hyaluronic acid uptake was increased by 50% after a 24-hr fast.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428725 TI - Liver regeneration after hepatic ischemia and reduced liver autotransplantation in the rat. AB - Current knowledge of liver regeneration after reduced liver transplantation is limited. Warm ischemia is one component of the reduced liver transplantation procedure that could have an impact on the regenerative response. To study this effect, we performed partial hepatectomy on male Long-Evans rats, with animals divided into four groups: group 1 underwent partial hepatectomy only; group 2 underwent partial hepatectomy and 40 min of ischemia; group 3 underwent partial hepatectomy, 40 min of ischemia and portocaval shunt surgery; and group 4 underwent partial hepatectomy and orthotopic autograft surgery. Group 5 consisted of sham-operated animals. Animals were killed 4, 24, 48, 72 and 96 hr after surgery. Thymidine kinase activity, mitotic index, a liver mass index and ornithine decarboxylase levels were used as parameters of liver regeneration. Aspartate transaminase was recorded. Maximal thymidine kinase and mitotic index were observed in group 1 animals at 24 hr. In groups 2, 3 and 4 maximal thymidine kinase activity and mitotic activity were observed 24 hr later at 48 hr. The magnitude of the peak response in these groups appeared to correlate with the duration of portal venous occlusion, with greatest increases occurring in those groups where portal stasis was most prolonged. The increase in liver mass for these groups was also delayed with respect to group 1 animals. The anticipated peak in ornithine decarboxylase levels was seen at 4 hr in group 1. The ornithine decarboxylase response in the other groups was disorganized, with delay of the recorded peaks.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428726 TI - Zonation of cholesterol and glycerolipid synthesis in regenerating rat livers. AB - Hepatic zonation of cholesterol and glycerolipid synthesis was investigated in regenerating rat livers 24 hr after partial hepatectomy. Tritiated acetate and [U 14C]glycerol were injected into rats' peritoneal cavities for a short-term labeling study. Periportal and perivenous hepatocytes were isolated by digitonin collagenase perfusion. Cholesterol synthesis was significantly higher in periportal hepatocytes of the sham-operated livers (periportal/perivenous = 1.67; p < 0.05). Twenty-four hours after partial hepatectomy, cholesterol synthesis was selectively decreased by 40% (p < 0.01) in periportal hepatocytes. Consequently, hepatic zonation of cholesterol synthesis was abolished in regenerating livers. To study the cholesterol homeostasis on a long-term basis, we substituted deuterated water (25% enriched) for drinking water for 5 days to label newly synthesized cholesterol in a steady state. This procedure clearly demonstrated the net negative cholesterol balance 24 hr after partial hepatectomy. However, the newly synthesized cholesterol contributed equally to the cellular cholesterol pool in both zones. The synthesis of glycerolipids, whether measured from tritiated acetate or [U-14C]glycerol, was significantly increased without apparent zonation in the regenerating livers (twofold increase in phospholipid, and threefold to sevenfold increase in triacylglycerol). We concluded that hepatic zonation of cholesterol synthesis is caused by higher de novo synthesis in periportal hepatocytes, which is abolished in regenerating livers. No zonation of glycerolipid synthesis exists in normal and regenerating livers. PMID- 8428727 TI - Stimulation of the collagen alpha 1 (I) endogenous gene and transgene in carbon tetrachloride-induced hepatic fibrosis. AB - Cirrhosis is characterized by a marked increase in the deposition of type I collagen and in the expression of the type I collagen genes alpha 1(I) and alpha 2(I). Although alpha 1(I) gene regulation has been extensively studied in cultured cells, these results may not be applicable to hepatic fibrogenesis in vivo. Therefore the regulation of the alpha 1(I) endogenous gene and an alpha 1(I) transgene was studied in a transgenic mouse model that has a single copy of a human alpha 1(I) gene segment containing the structural gene and 1.6 Kb of 5' DNA and 20 Kb of 3' DNA. To initiate hepatic fibrogenesis, we treated mice with the hepatotoxin carbon tetrachloride, either in a single dose or in biweekly doses for a period of 3 to 8 wk. Subsequently, hepatic alpha 1(I) messenger RNA levels were determined by a species-specific RNase protection assay. Carbon tetrachloride injections coordinately increased the messenger RNA levels of the alpha 1(I) endogenous gene and the transgene, both immediately and after 8 wk. These experiments demonstrate that this alpha 1(I) transgene fragment contains information sufficient for appropriate basal and carbon tetrachloride-stimulated hepatic expression. They further demonstrate that sufficient homology exists between the human and mouse regulatory elements for the recognition of human cis acting elements by mouse trans-acting factors. Thus transgenic mice provide a unique model in which to characterize the collagen alpha 1(I) regulatory elements that are required in vivo for pathophysiological responses. PMID- 8428728 TI - Clearance by the liver in cirrhosis. I. Relationship between propranolol metabolism in vitro and its extraction by the perfused liver in the rat. AB - To delineate the factors responsible for impaired clearance in cirrhosis, we examined propranolol disposition in rats with carbon tetrachloride-induced cirrhosis and compared it with that in control animals, rats treated with chlorpromazine (an inhibitor of propranolol metabolism) and rats with acute liver injury. We measured the extraction ratio of propranolol by the isolated perfused liver and related it to estimates of propranolol drug-metabolizing enzyme activity in homogenates of the same livers. In control animals, drug-metabolizing enzyme activity (measured as the ratio Vmax/Km) averaged 5,319 +/- 1,193 ml/min; the extraction ratio in the perfused liver was close to 1.0 (0.97 +/- 0.01). Important decreases of microsomal enzyme activity were observed in rats treated with chlorpromazine (30 +/- 27 ml/min, p < 0.001) and in rats with acute liver injury (724 +/- 401 ml/min, p < 0.001), accounting for the decrease in the hepatic extraction ratio by the perfused liver (0.33 +/- 0.09 and 0.71 +/- 0.04, respectively, p < 0.01). In cirrhotic livers, enzyme activity was not significantly different from that of controls (3,592 +/- 1,857 ml/min) and could not account for the observed decrease in extraction (0.66 +/- 0.14, p < 0.01). The extraction of antipyrine by the isolated perfused liver was also measured as an index of microsomal enzyme activity and related to propranolol extraction. Antipyrine extraction was decreased by 90% in acute liver injury, compared with 33% in cirrhosis, suggesting a much greater reduction of microsomal enzyme activity in the former group.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428729 TI - Hepatic fibrosis as a predictor of hepatic regenerative activity after partial hepatectomy in the rat. AB - Liver regeneration is an essential component of the recovery period after partial hepatectomy. Unfortunately, tests that accurately predict regenerative activity in the postoperative period have yet to be described. This study was designed to determine whether the extent of hepatic fibrosis correlates with liver regeneration activity after partial hepatectomy in rats with carbon tetrachloride induced liver disease. Two groups of adult male Sprague-Dawley rats (12 to 30/group) were treated for 20 to 22 wk with weekly intragastric doses of carbon tetrachloride or vehicle. All rats then underwent 70% hepatectomy while under ether anesthesia. Liver regeneration activity was determined at 24 and 48 hr by [3H]thymidine incorporation into DNA. Hepatic fibrosis was calculated at the time of partial hepatectomy by automated image analysis on Van Gieson-stained liver tissue. Although a significant inverse correlation was found between the extent of hepatic fibrosis and DNA synthesis when all rats were considered (carbon tetrachloride-treated and vehicle-treated) at 24 and 48 hr after partial hepatectomy (r = -0.4943 and -0.7396, respectively; p < 0.05), no such correlation existed when carbon tetrachloride-treated rats were considered independently (r = -0.3231 and -0.0910 at 24 and 48 hr, respectively). In conclusion, we believe that in diseased livers, preoperative quantitation of hepatic fibrosis on automated image analysis does not serve as a useful predictor of liver regeneration activity. PMID- 8428730 TI - Morphometry of the liver after liver transplantation in the rat: significance of an intact arterial supply. AB - Two models of orthotopic liver transplantation in the rat currently used are with and without reconstruction of the hepatic artery. The aim of this study was to assess the importance of the arterial blood supply to liver structure after orthotopic liver transplantation with advanced morphometric methods. Orthotopic liver transplantation was performed in male Lewis rats, of which 11 underwent reconstruction of the hepatic artery and 11 did not. A group of untreated controls (n = 5) and a group of animals with hepatic artery ligation (n = 4) were included. Eight weeks after surgery liver tissue was harvested and subjected to systematic random sampling. A point-counting method was used, volume fractions of tissue components were determined. Liver samples from rats that underwent hepatic artery reconstruction had preserved lobular architecture, and estimated bile duct (0.71% +/- 0.33% [S.D.]) and connective tissue (1.89% +/- 0.52%) volumes were not significantly different from those of controls (bile duct, 0.34% +/- 0.17%; connective tissue, 0.70% +/- 0.07%). In contrast, liver samples from rats that did not undergo hepatic artery reconstruction showed bile duct proliferation (7.19% +/- 4.83%; p < 0.05) and an increase in connective tissue volume (7.54% +/ 3.68%; p < 0.05) associated with a decrease in hepatocyte volume (controls, 87.3% +/- 0.3%; rats that underwent arterialization, 85.5% +/- 1.0%; rats that did not undergo arterialization, 73.2% +/- 8.2% [p < 0.05]). Interestingly, hepatic artery ligation had no significant effect on any parameter.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428731 TI - The biology of the bile canaliculus, 1993. PMID- 8428732 TI - ATP-dependent transport systems for organic anions. PMID- 8428733 TI - The self-sustained sequence replication amplification system--move over, PCR. PMID- 8428734 TI - Policy ... use of emergency departments. PMID- 8428735 TI - Holding networks together. Shared information will be glue for reformed health system. PMID- 8428736 TI - Military maneuvers. DoD moves ahead with integrated clinical records. PMID- 8428737 TI - All connected. Infrastructure is path to IS growth. PMID- 8428738 TI - HELP (health evaluation through logical processing) on the way. Clinical system lays framework for CPR. PMID- 8428739 TI - The real thing. Future information needs will require 'true' hospital CIOs. PMID- 8428741 TI - Medical execs rising. Today's VPMAs (vice president of medical affairs) are high profile and strategy savvy. PMID- 8428740 TI - Caring for the poor--and more. Public hospitals prepare for a changed delivery system. PMID- 8428742 TI - Homeward bound. Hospitals see solid future for home health care. PMID- 8428743 TI - Data watch. Survey outlines hospital collaboration efforts. PMID- 8428744 TI - ProPAC's advice: help beneficiaries, spare providers. PMID- 8428745 TI - COBRA waters look no clearer after recent reversal. Johnson v. University of Chicago Hospitals. PMID- 8428747 TI - Primary care physicians still in demand. PMID- 8428746 TI - Mind & body medicine. A new paradigm? AB - Innovative techniques to enhance patient healing are being explored, and the term used to describe them is "mind/body medicine." What's new about the phenomenon? Traditional physicians are taking a closer look. PMID- 8428748 TI - Medical staff development: board role crucial. PMID- 8428749 TI - Prenatal sex determination from maternal peripheral blood using the polymerase chain reaction. AB - We have investigated the use of a nested polymerase chain reaction assay for the detection of a fetal-specific Y-chromosomal sequence (DYS14) from DNA extracted from unsorted maternal peripheral blood. Serial dilutions of male DNA into female cord blood DNA indicated that the assay could detect an equivalent of a single male cell in 300,000 female cells. The assay exhibited absolute specificity for male DNA with no amplification from a DNA panel obtained from 10 female cord blood samples. When used on DNA extracted from unsorted peripheral blood from a series of pregnant women, the predictive values of a positive test for a male fetus were 86%, 67% and 87% in the first, second and third trimesters, respectively. We have also demonstrated that retesting the samples allows the detection of a proportion of male-bearing pregnancies with a high degree of accuracy, in that all 15 women who gave positive signals in two consecutive amplifications had male fetuses. We have also applied the test at 8 weeks postpartum to eight women who had previously delivered male babies; no Y-specific signal could be detected in any of them, suggesting that most women have cleared their circulation of fetal cells by 8 weeks after parturition. PMID- 8428750 TI - Localization of the intronless gene coding for calmodulin-like protein CLP to human chromosome 10p13-ter. AB - The functional intronless gene coding for a calmodulin-like protein (CLP) has been localized to human chromosome 10p13-ter. Chromosomal assignment was performed by Southern blot analysis of DNA from human-rodent somatic cell hybrids and amplification of a CLP gene-specific 1090-bp DNA fragment by the polymerase chain reaction (PCR) on DNA from human-hamster cell hybrids. Chromosomal sublocalization was carried out by in situ hybridization of human chromosome metaphase spreads. The CLP gene is the first member of the human calmodulin/calmodulin-like gene family to be chromosomally sublocalized. Its presence near the telomeric end of the short arm of chromosome 10 may be of significance with respect to its highly (epithelial) cell-type restricted expression in vivo and strong downregulation upon malignant transformation. The generation of a human CLP gene-specific sequence tag site specified by the two primers used for PCR should prove useful for future linkage studies. PMID- 8428752 TI - Chromosomal analysis of unfertilized human oocytes prepared by a gradual fixation air drying method. AB - Two hundred and sixty-five unfertilized human metaphase II (M II) oocytes from an in vitro fertilization program were studied cytogenetically using our chromosomal technique, a gradual fixation-air drying method. Of the 265 oocytes, 185 (70%) were successfully karyotyped. There were 21 aneuploids (11.4%) consisting of 8 hyperhaploids (4.3%), 11 hypohaploids (5.9%) and 2 complex cases (1.1%). There were also 9 structural anomalies (4.9%) and 18 diploids (9.7%). In aneuploidy, the loss or gain of dyads (so-called nondisjunction) occurred more frequently than the loss or gain of monads (so-called predivision). The frequency of abnormally behaved chromosomes (segregation errors) due to nondisjunction, anaphase lag and predivision was studied among the seven chromosomal groups (A-G) and compared with the frequency expected from an equal probability of segregation errors in each of the 23 chromosomes. The observed frequency was somewhat higher than the expected frequency in groups E and G but the difference was not statistically significant in either group. These results were discussed in relation to previous studies on human M II oocyte chromosomes. PMID- 8428751 TI - Decreased extracellular deposition of fibrillin and decorin in neonatal Marfan syndrome fibroblasts. AB - Abnormalities of the microfibrillar protein fibrillin (Fib) have been reported in Marfan syndrome (MFS). The so-called neonatal Marfan syndrome (nMFS) is a lethal phenotype displaying features that are not seen in classical MFS. We have therefore studied the biosynthesis and extracellular deposition of Fib and decorin in fibroblasts from a patient with nMFS and controls. Immunofluorescence of the patient's cell cultures showed an almost complete absence of Fib and a marked reduction of decorin in the extracellular matrix (ECM). The nMFS skin revealed Fib on subbasal microfibrillar bundles in the papillary dermis, and Fib associated with elastic fibers in the reticular dermis; the bundles and fibers were fragmented and thinner than normal. Pulse-chase labeling of cells with [35S]Met/Cys revealed moderately reduced secretion, but a diminished deposition of Fib in the ECM; this was more apparent at a longer chase time. Fib mRNA and synthesis appeared to be normal, whereas both decorin mRNA and biosynthesis were reduced. We therefore assume a structural Fib defect in this patient causing reduced deposition into and/or enhanced removal from the ECM, whereas the reduced decorin biosynthesis may be a secondary regulatory phenomenon. The clinical relevance of this remains unclear. Our findings imply that Fib defects may be responsible for the severe, complex phenotype of nMFS. PMID- 8428753 TI - The human interleukin-6 receptor alpha chain gene is localized on chromosome 1 band q21. AB - The human interleukin-6 receptor alpha chain (IL6R alpha) gene was regionally mapped to chromosome 1 band q21 by fluorescence in situ hybridization. As hybridization probes, partially overlapping lambda clones encompassing 28 kb of the genomic region of the gene were used. These clones were isolated using a polymerase chain reaction (PCR)-generated fragment of the 3' non-coding region of the gene. This localization confirms and extends the provisional assignment of the IL6R alpha gene to chromosome 1, for which a panel of somatic cell hybrids was used. PMID- 8428754 TI - Exclusion of two candidate pigment loci, c and b, part of chromosome 11p, and 33 random polymorphic markers as the locus for tyrosinase-positive oculocutaneous albinism. AB - The locus for Tyrosinase-Positive Oculocutaneous Albinism (ty-pos OCA) has not yet been localised. The search for the ty-pos OCA locus has included a search for linkage to candidate pigment loci and a candidate chromosomal region, as well as a random search using highly polymorphic markers in 42 families, including 271 individuals of whom 79 are affected. The lod scores for the tyrosinase (TYR) locus (11q14-q21), homologous to the albino locus, c, in the mouse and the CAS2/TRP1 locus (9p22-pter), homologous to the brown locus, b, in the mouse were 5.89 and -7.22, respectively, at a recombination fraction of theta = 0.01, thus excluding them from being the ty-pos OCA locus. In the candidate chromosomal region, 11p, four loci (probes) were tested, SAA (pSAA82), CALC (pHC36), HBB (Gamma-globin haplotype) and an AC repeat polymorphism at the Wilm's Tumour locus (WT1). A portion of 11p was excluded with the following lod scores: pSAA82 lod = 2.05 at theta = 0.10; pHC36 lod = -3.87 at theta = 0.05; gamma-globin haplotype lod = -2.80 at theta = 0.10; and WT1 lod = -2.34 at theta = 0.10. Thirty-three polymorphic markers randomly distributed on 13 different chromosomes were all excluded from close linkage to ty-pos OCA. PMID- 8428755 TI - Apolipoprotein B signal peptide polymorphism in patients with myocardial infarction and controls. "The ECTIM study". AB - We report the allele frequencies of the apolipoprotein B (Apo B) signal peptide polymorphism in patients with myocardial infarction and compare them with controls. The first sample consists of 197 myocardial infarction patients and 168 controls from Belfast (UK). The second sample consists of 167 myocardial infarction patients and 205 controls from Strasbourg (France), and the third consists of 71 patients and 146 controls from Haute-Garonne (Toulouse, France). No significant differences were observed in the frequency distribution of genotypes among cases and controls or between populations. However, there were more rare homozygotes in the Belfast cases. Significant associations were observed between the Apo B signal peptide polymorphism and mean levels of total cholesterol, low density lipoprotein cholesterol, Apo B and lipoprotein particles containing Apo (a) [Lp(a)] in the Strasbourg control population. Individuals homozygous for the rare allele had higher levels of these lipid parameters. In Belfast, although not statistically significant, the Apo B signal peptide polymorphism had a similar effect on Apo-B-related parameters as seen in Strasbourg. No significant associations were observed in the Haute-Garonne population where the risk of myocardial infarction is three times lower than in Belfast. In all three populations, the average Lp(a) levels were consistently different among Apo B signal peptide genotypes. These data implicate the Apo B signal peptide in determining some of the risks of myocardial infarction in these populations. Regardless of the exact mechanism, the Apo B signal peptide is an important candidate locus for the study of potentially atherogenic lipid variants. PMID- 8428756 TI - A study of genetic linkage heterogeneity in 35 adult-onset polycystic kidney disease families. AB - A genetic heterogeneity analysis of 35 kindreds with adult-onset polycystic kidney disease (ADPKD) was carried out using the D16S85, D16S84, D16S125 and D16S94 loci that are closely linked to the PKD1 locus on chromosome 16. The results show that the likelihood of two ADPKD loci is 2,514.9 times greater than for a single locus (P < 0.0001). The maximum likelihood lod score is 27.38 under heterogeneity with PKD1 lying 4.9 cM proximal to D16S85 (in males). At least 3% of kindreds are unlinked to PKD1, since the 95% confidence limits of alpha, the proportion of families linked to PKD1, are 0.54-0.97. Only 2 out of 35 kindreds (5.7%) show statistically significant evidence of non-linkage to PKD1, with conditional probabilities of 0.987 and 0.993 that the disease locus is unlinked. This confirms the existence of a small subgroup of ADPKD kindreds that are unlinked to PKD1 and provides a firm basis for genetic counselling of this population on the basis of DNA probes. PMID- 8428758 TI - Establishing credibility in a new nursing job. PMID- 8428757 TI - Is this the job for you? PMID- 8428759 TI - Returning to school: guide for the RN consumer. PMID- 8428760 TI - Five strategies to survive your first interview. PMID- 8428761 TI - Writing a resume. PMID- 8428762 TI - Nursing organizations. PMID- 8428763 TI - The nurse I admire most. PMID- 8428764 TI - Preparing for NCLEX-RN. PMID- 8428765 TI - Non-human primate MHC class I sequences, 1992. PMID- 8428766 TI - The mouse Eb meiotic recombination hotspot contains a tissue-specific transcriptional enhancer. AB - A meiotic recombination hotspot exists within the second intron of the mouse major histocompatibility complex (MHC) gene, Eb. In the present study, a small fragment from the intron which contains two potential transcriptional regulatory elements was cloned into an expression vector and its effect on transcription was tested. This fragment was found to contain tissue-specific transcriptional enhancer activity. An octamer-like sequence and a B motif may contribute to this enhancer activity. Similar regulatory sequences with the same orientation and distance from one another are found in another mouse MHC recombination hotspot. PMID- 8428767 TI - Evolutionary relationships of the classes of major histocompatibility complex genes. AB - A number of hypotheses have been proposed to account for the evolutionary origin of the classes of major histocompatibility complex (MHC) genes of vertebrates. According to one hypothesis the class II MHC evolved first, whereas another hypothesis holds that the class I MHC originated first as a result of a recombination between an immunoglobulin-like C-domain and the peptide-binding domain of an HSP70 heat-shock protein. A phylogenetic tree of C-domains from MHC and related molecules supports a relationship between the class II MHC alpha chain and beta 2-microglobulin and between the class II MHC beta-chain and the class I alpha chain. If this phylogeny is correct, the hypothesis that the class I MHC evolved by recombination with HSP70 is less parsimonious than the hypothesis that class II evolved first. Furthermore, when MHC peptide-binding domains are simultaneously aligned with HSP70 domains and with V-domains from members of the immunoglobulin superfamily, they are slightly more similar to the latter than to the former; and the class II alpha 1 and beta 1 domains show much greater similarity to each other than would be expected if they evolved from separate HSP70 domains. Thus, most evidence supports the hypothesis that the ancestral MHC molecule had a class II-like structure. PMID- 8428768 TI - Familial clustering of IGHC deletions and duplications: functional and molecular analysis. AB - The human immunoglobulin heavy chain constant region locus (IGHC) comprises nine genes and two pseudogenes clustered in a 350 kilobase (kb) region on chromosome 14q32. Several IGHC haplotypes with single or multiple gene deletions and duplications have been characterized. The most likely mechanism accounting for these unusual haplotypes is the unequal crossing-over between homologous regions within the locus. Here we report the analysis of an unusual case of familial clustering of deletions/duplications. In the two branches of the BON family, three duplicated and two deleted haplotypes, all probably independent in origin, have been characterized. The structure of the haplotypes, one of which is described here for the first time, supports the hypothesis of homologous unequal crossing-over as the origin of recombinant haplotypes. The analysis of serological markers in a subject carrying one deleted and one duplicated haplotype allowed us the first direct inferences concerning the functions of the duplicated IGHC haplotypes. PMID- 8428769 TI - Structure and evolution of the promoter regions of the DQA genes. AB - HLA-DQ antigens are unique among the class II antigens in that their alpha chains are highly polymorphic. In the present study, we characterized the general structure of the promoter regions of the DQA genes derived from different DR haplotypes and defined their nucleotide sequence polymorphisms. The promoter of each DQA1 allele contains three sequence motifs which are not present in non-DQA related class II genes: one identical to a tumor necrosis factor (TNF alpha) response element, one similar to an NF kappa B binding element, and one similar to a W motif. All DQA alleles lack TATA and CCAAT boxes in the proximal promoter region but carry other sequence elements characteristic of MHC class II genes, including S, X, X2, and Y boxes, and a pyrimidine-rich tract upstream of the X box. Nucleotide sequence polymorphisms among the various DQA1 alleles were noted within the promoter region, with some of the differences mapping within, or close to, regulatory elements that are important for the expression of MHC class II genes. All DQA1 alleles carry an unrearranged, full length, Alu-Sx related repeat immediately upstream of the proximal promoter region. This repeat was not present in the DQA2 (DXA) genes analyzed, confirming that DQ locus duplication probably occurred before integration of the Alu repeat into the primordial DQA1 locus, some 31-43 million years (myr) ago. The DQA2 promoter region is highly conserved between DR4 and DR3 haplotypes, with the degree of conservation exceeding that expected from the neutral mutation rate. PMID- 8428771 TI - Analysis of B-L beta-chain gene expression in two chicken cDNA libraries. PMID- 8428770 TI - Alleles and haplotypes of the MHC-encoded ABC transporters TAP1 and TAP2. AB - TAP1 and TAP2 are two major histocompatibility complex (MHC) genes, located between HLA-DP and -DQ, whose products form a transporter molecule involved in endogenous antigen processing. Polymorphic residues have been described in both genes and, in this study, we have identified another polymorphic site within the adenosine triphosphate (ATP)-binding domain of TAP2. We have used the amplification refractory mutation system (ARMS) polymerase chain reaction (PCR) to characterize TAP1 and TAP2 alleles and haplotypes in a reference panel of 115 homozygous typing cell lines. Of four possible TAP1 alleles, we observed three, and of eight possible TAP2 alleles, we observed five. Among 88 (homozygous typing cells) (HTCs) homozygous at HLA-DR, -DQ and -DP, 80 were also homozygous at TAP1 and TAP2. Of 27 HTCs homozygous at HLA-DR and -DQ, but heterozygous at -DP, 14 were homozygous at TAP1 or TAP2 and 13 heterozygous, consistent with recombination taking place either side of the TAP loci. Of the fifteen possible combinations of TAP1 and TAP2 alleles, we observed eleven, each at a frequency similar to that predicted by individual allele frequencies. In this ethnically heterogenous panel there is no indication that particular combinations of TAP1 and TAP2 have been maintained together. PMID- 8428772 TI - Fine map of a region homologous to the rat grc complex in the TL region of the mouse. PMID- 8428773 TI - Tissue-specific RNA processing for the complement C4 gene transcript in the H-2k mouse strain. PMID- 8428774 TI - Localization of a new gene adjacent to the HSP70 genes in human and mouse MHCs. PMID- 8428775 TI - Skeletal muscle blood flow. A possible link between insulin resistance and blood pressure. AB - Insulin resistance has recently been found to be a common feature of essential hypertension. We have tested the hypothesis that reduced skeletal muscle blood flow in response to insulin may at least partially account for the wide range of insulin sensitivity observed in normotensive subjects. To this end, we studied 19 lean (body mass index < or = 27) subjects exhibiting basal mean arterial pressures ranging from 58 to 110 mm Hg. All subjects were normotensive with the exception of one. Each subject was studied at baseline and during a hyperinsulinemic (600 milliunits/m2 per minute) euglycemic clamp to quantitate insulin sensitivity. Mean arterial pressure was monitored invasively, and both leg (muscle) blood flow and cardiac output were measured by indicator dilution techniques, allowing the determination of both systemic and leg (or muscle) vascular resistance. In response to hyperinsulinemia, both cardiac output and leg blood flow increased approximately 37% and 80% (p < 0.01), respectively. Rates of insulin-mediated glucose uptake were inversely correlated with the baseline mean arterial pressure (r = -0.62, p < 0.01). The individual increment in leg blood flow above baseline in response to insulin was inversely proportional to the height of the baseline mean arterial pressure (r = -0.59, p < 0.01). Mean arterial pressure and insulin-mediated glucose uptake were not correlated with either age or body fat content.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428776 TI - The vasodilator action of insulin. Implications for the insulin hypothesis of hypertension. PMID- 8428777 TI - Myotrophin induces early response genes and enhances cardiac gene expression. AB - We have identified and partially sequenced a soluble factor, myotrophin, from spontaneously hypertensive rat hearts and hypertrophic human hearts that enhances myocyte protein synthesis and stimulates myocardial cell growth. Our studies suggest that myotrophin may be a biochemical link between hemodynamic stress and myocardial cellular hypertrophy. When rat neonatal cardiac myocytes maintained in culture were incubated with myotrophin for 30 minutes, they showed a marked increase in c-myc, c-fos, and c-jun messenger RNA levels. Cardiac myocytes treated for 24 hours with myotrophin showed a fourfold increase in connexin 43 (gap junction protein), a sixfold increase in atrial natriuretic factor, a threefold increase in skeletal alpha-actin, and a threefold increase in total myosin transcript levels. Studies on myosin isoforms showed a selective increase in the beta-myosin heavy chain transcript levels but no reciprocal decrease in alpha-myosin heavy chain transcript levels. Our data suggested that myotrophin appears to be a primary modulator for myocardial cell growth and differentiation and may play an important role in the pathogenesis of cardiac hypertrophy. Myotrophin may be involved in the upregulation of myofibrillar protein and the activation of cardiac gene transcription during growth and hypertrophy of the myocardium, and the induction of early response gene expression may be linked to this response. PMID- 8428778 TI - Bradykinin peptides in kidney, blood, and other tissues of the rat. AB - The bradykinin peptide system is a tissue-based system with potent cardiovascular and renal effects. To investigate the regulation of this system, we developed a highly sensitive amino terminal-directed radioimmunoassay that, with high performance liquid chromatography, enables the measurement of bradykinin-(1-7), bradykinin-(1-8), and bradykinin-(1-9). Together with a carboxy terminal-directed radioimmunoassay, we characterized bradykinin peptides in rat kidney and blood. The predominant bradykinin peptides in kidney were bradykinin-(1-9) (approximately 100 fmol/g wet weight of tissue) and bradykinin-(1-7) (approximately 70 fmol/g), with low levels of bradykinin-(1-8) (approximately 8 fmol/g) and bradykinin-(4-9) (approximately 12 fmol/g) detectable; bradykinin-(2 9) and bradykinin-(3-9) were below the limits of detection. In blood, the levels of bradykinin-(1-9) were very low (approximately 2 fmol/ml), and other bradykinin peptides were below the limits of detection. Ile,Ser-bradykinin and Met,Ile,Ser bradykinin were below the limits of detection in both kidney and blood, indicating that T-kininogen makes no detectable contribution to renal or circulating bradykinin peptides. Administration of the angiotensin converting enzyme inhibitor perindopril was associated with an approximate twofold increase in renal levels of bradykinin-(1-8) and bradykinin-(1-9) and a decrease in the bradykinin-(1-7)/bradykinin-(1-9) ratio. The amino terminal-directed radioimmunoassay was also applied to heart, aorta, brown adipose tissue, adrenal lung, and brain. For these tissues, bradykinin-(1-7) and bradykinin-(1-9) were of similar abundance (16-340 fmol/g), with lower levels of bradykinin-(1-8). These studies demonstrate that tissue levels of bradykinin peptides are much higher than circulating levels, consistent with their formation at a local tissue site. Of peptides derived from K-kininogen, bradykinin-(1-9) is the predominant bioactive peptide in all tissues, and a major pathway of bradykinin-(1-9) metabolism involves the formation of bradykinin-(1-7). In kidney, angiotensin converting enzyme plays an important role in bradykinin-(1-9) metabolism, and increased bradykinin-(1-9) and bradykinin-(1-8) levels may mediate in part the renal effects of converting enzyme inhibition. PMID- 8428779 TI - Atrial natriuretic peptide blunts the cellular effects of cyclosporine in smooth muscle. AB - The effect of cyclosporine A to enhance vasoconstrictor-induced calcium (Ca2+) mobilization in vascular smooth muscle cells may contribute to important side effects in cyclosporine therapy such as hypertension and nephrotoxicity. On the other hand, atrial natriuretic peptide (ANP) is known to diminish vasoconstrictor stimulated Ca2+ mobilization. The present study, therefore, examined the interaction of cyclosporine and ANP on Ca2+ kinetics in cultured rat vascular smooth muscle cells. Intracellular free calcium concentrations ([Ca2+]i) were measured using fura-2. 45Ca2+ was used to estimate Ca2+ efflux and cellular Ca2+ influx. Preincubation of the cells with cyclosporine (10 micrograms/ml) for 12 minutes lowered basal [Ca2+]i from 48 +/- 4 to 28 +/- 3 nM (p < 0.01). However, in the presence of cyclosporine, the angiotensin II (10(-8) M)-stimulated rise of [Ca2+]i was increased from 296 +/- 22 to 460 +/- 47 nM (p < 0.001). ANP (5 x 10( 9) M) blocked the Ca2+ mobilization by angiotensin II (71 +/- 7 versus 69 +/- 7 nM, NS) and also completely inhibited the effect of angiotensin II in the presence of cyclosporine (77 +/- 5 versus 78 +/- 5 nM, NS). Basal efflux as well as angiotensin II-stimulated 45Ca2+ efflux were not altered by preincubation with cyclosporine, indicating that the effect of cyclosporine on [Ca2+]i was not due to an inhibition of 45Ca2+ efflux.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428780 TI - Regional angiotensin II production in essential hypertension and renal artery stenosis. AB - To study regional metabolism and production of angiotensin II, we measured steady state plasma levels of 125I-angiotensin I and II and endogenous angiotensin I and II in the aorta and the antecubital, femoral, renal, and hepatic veins during systemic infusion of 125I-angiotensin I or II. Extraction of arterially delivered angiotensin II ranged from 30-50% in the limbs to 80-100% in the renal and hepatomesenteric vascular beds both in essential hypertension (n = 13) and in unilateral renal artery stenosis (n = 7). Across the limbs, 20-30% of arterially delivered angiotensin I was converted to angiotensin II in both groups, and there was no arteriovenous gradient in endogenous angiotensin II. No conversion of arterially delivered angiotensin I was detected across the renal and hepatomesenteric beds, and there was net extraction of angiotensin II from the systemic circulation by these beds. Although regional production of angiotensin I at tissue sites made a significant contribution to its level in the veins, little of this locally produced angiotensin I reached the regional veins in the form of angiotensin II, even in the kidney with artery stenosis, where the venous levels of locally produced angiotensin I were particularly high. These results provide no evidence for a source of circulating angiotensin II other than blood-borne angiotensin I and illustrate the high degree of compartmentalization of angiotensin I and II production. PMID- 8428781 TI - Vascular smooth muscle cell calcium fluxes. Regulation by angiotensin II and lipoproteins. AB - This study examined 45Ca uptake, 45Ca efflux, and the distribution of exchangeable 45Ca in confluent, quiescent cultures of aortic smooth muscle cells (VSMCs) from normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). These parameters were investigated under basal conditions and after addition of angiotensin II (Ang II) and low (LDL) and high (HDL) density lipoproteins. Basal 45Ca uptake was approximately 50% greater in VSMCs from SHRs (p < 0.005 versus WKY). Calcium antagonists (diltiazem or nifedipine) abolished this difference. The 45Ca uptake response to Ang II was approximately twofold greater in SHR than in WKY VSMCs (p < 0.05), and Ang II-induced increments of 45Ca uptake were weakly inhibited (by approximately 15-25%) by calcium antagonists. Lipoproteins also stimulated 45Ca uptake in VSMCs, and the apparent affinity of this process was approximately fivefold greater for LDL than for HDL. Calcium antagonists did not inhibit either LDL- or HDL-induced 45Ca uptake. SHR and WKY VSMCs did not differ with respect to 45Ca uptake induced by either LDL or HDL. The initial size of the slowly exchangeable pool of intracellular Ca2+ was approximately 35% greater in SHR VSMCs (p < 0.05 versus WKY). Ang II-induced mobilization of intracellular calcium (measured as the decrease in 45Ca content of the slowly exchangeable pool) was threefold greater in SHR VSMCs (p < 0.005 versus WKY). LDL and HDL marginally stimulated 45Ca efflux from this pool (< or = 20% above control) and to comparable extents in both SHR and WKY VSMCs.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428782 TI - Guanine nucleotide regulatory proteins in the spontaneously hypertensive rat. AB - We compared guanine nucleotide regulatory protein (G protein) levels and function in plasma membranes from resistance vessels (mesenteric arteries) isolated from spontaneously hypertensive (SHR) and normotensive Wistar rats. G protein function was deduced from studies of adenylate cyclase activity. Although the basal level of adenylate cyclase activity (+/- Mn2+ ions) was significantly greater in SHR membranes, addition of agents that function via the stimulatory G protein--i.e., NaF (10(-2) M), (-)-isoproterenol (10(-4) M), and prostaglandin E1 (10(-5) M)- resulted in a significantly lower stimulatory response in SHR membranes. Ligands that function via the inhibitory G protein--i.e., adrenaline (10(-5) M)/propranolol (10(-5) M) (this combination being equivalent to an alpha 2 receptor agonist), carbachol (10(-3) M), and serotonin (10(-5) M)--were responsible for only slight inhibitory responses in both SHR and Wistar rat membranes, which were not significantly different. Western blotting identified the presence of Gs, Gi2, and Gi3 alpha-subunits in rat vascular smooth muscle, but there were no differences in the levels of these G protein alpha-subunits found in SHR and Wistar rat plasma membranes. The levels of the beta-subunit in the two sets of membranes were also similar. In conclusion, there is a reduced response in adenylate cyclase activity to agents that function via the stimulatory G protein in SHR membranes. However, this is not a consequence of altered levels of the different G protein subunits. PMID- 8428783 TI - Coronary vascular response to the cerebral ischemia reflex. AB - Most centrally mediated sympathoexcitatory reflexes produce increases in arterial pressure, heart rate, and peripheral vascular resistance, including coronary vasoconstriction. Cerebral ischemia also causes large increases in arterial pressure and peripheral vasoconstriction but with modest or variable changes in heart rate. To examine the effect of cerebral ischemia on coronary vascular resistance, we produced cerebral ischemia in 14 cats by occluding the right brachiocephalic and left subclavian arteries for 30 seconds. After vagotomy and beta-blockade, a marked increase in arterial pressure (89 +/- 14%) and coronary vascular resistance (52 +/- 7%) was seen. After inhibition of the carotid baroreceptor reflex by surgical denervation and application of topical lidocaine, the increase in arterial pressure to cerebral ischemia was not affected, but the increase in coronary vascular resistance was attenuated (33 +/- 6%; p < 0.05 versus before denervation) to a level expected with autoregulation. To evaluate the possible contribution of the chemoreflex on coronary blood flow during cerebral ischemia, we conducted separate experiments in which nicotine was injected into both carotid arteries. Coronary constriction was not observed. Adrenalectomy and upper extremity ischemia likewise did not alter coronary vascular resistance. We conclude that cerebral ischemia elicits neurally mediated coronary vasoconstriction as a result of baroreceptor hypotension rather than directly. The relative absence of neurogenic coronary constriction and changes in heart rate suggest that sympathoexcitation during cerebral ischemia is directed more toward the peripheral vasculature than the heart. PMID- 8428785 TI - Altered blood pressure during sleep in normotensive subjects with type I diabetes. AB - This study was designed to examine the circadian pattern of blood pressure in children and young adults with type I diabetes who were completely normotensive by standard criteria. Forty-five patients and the same number of age- and sex matched control subjects were studied. In diabetic children of 10-14 years of age, the nocturnal fall in systolic and diastolic blood pressures was intact. In diabetics of 15-20 years of age, the fall in systolic blood pressure was blunted; in diabetics of 21-37 years of age, the fall in both systolic and diastolic blood pressures during sleep was blunted. When data from all diabetic subjects were pooled and analyzed in a multiple linear regression model, mean blood pressure during sleep correlated best with urinary albumin excretion (r = 0.60). On the basis of this finding, we subdivided our patients into two groups: a microalbuminuric group (urinary albumin excretion > 30 mg per 24 hours; mean, 160.3 +/- 29.7; n = 11) and a normoalbuminuric group (urinary albumin excretion < 30 mg per 24 hours; mean, 6.6 +/- 6.5; n = 34). Both systolic and diastolic blood pressures during sleep were higher in microalbuminuric (121.1 +/- 3.3 and 69.3 +/ 2.5 mm Hg, respectively) than in normoalbuminuric diabetics (114.2 +/- 1.8 and 60.1 +/- 1.2 mm Hg, p < 0.05) or control subjects (113.3 +/- 1.2 and 60.1 +/- 1.2 mm Hg, p < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428784 TI - Cardiac volume receptor reflex in borderline hypertensive rats. AB - With increased dietary NaCl intake (8% NaCl), borderline hypertensive rats (BHR) develop hypertension and exhibit an exaggerated natriuresis in response to intravenous isotonic saline volume expansion. The exaggerated natriuresis is mediated by the concurrent exaggerated withdrawal of efferent renal sympathetic nerve activity since prior renal denervation eliminates the exaggerated natriuretic response. It was the objective of the present study to examine cardiac volume receptor reflex control of efferent renal sympathetic nerve activity in BHR made hypertensive by increased dietary NaCl intake. BHR were fed either 1% or 8% NaCl from age 4 to 16 weeks. BHR fed 8% NaCl were hypertensive (148 +/- 9 mmHg) compared with BHR fed 1% NaCl (115 +/- 6 mm Hg, p < 0.05). In one protocol, measurements of right atrial pressure and efferent renal sympathetic nerve activity were made in sinoaortic-denervated BHR before and during a 10% body weight intravenous isotonic saline volume load. Compared with 1% NaCl BHR, 8% NaCl BHR showed both a greater maximal inhibition of efferent renal sympathetic nerve activity (-67 +/- 4% versus -31 +/- 3% of control, p < 0.05) and gain (-22.0 +/- 2.3 versus -9.7 +/- 1.7%/mm Hg, p < 0.05). In a second protocol, measurements of efferent renal sympathetic nerve activity were made in sinoaortic-denervated BHR before and during graded frequency stimulation of the central portion of the sectioned vagus nerve.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428786 TI - Mild hypovolemic stress alters autonomic modulation of heart rate. AB - In response to changes in central venous volume, changes in vagal efferent cardiac outflow have been demonstrated in animals but not in humans. In this study, frequency domains analysis was used to quantify modulation of heart rate by respiration and blood pressure in normal human adults undergoing mild central hypovolemic stress induced by blood donation and postural change. In supine subjects, blood donation caused no change in mean heart rate, pulse pressure, or in the variance of heart rate or blood pressure. There were small decreases in mean and systolic blood pressure. A significant decrease in vagal modulation of heart rate was seen in the 0.12-0.5-Hz frequency band, as measured by the change in the relation of lung volume to heart rate in this frequency band (-4.49 beats per minute [pbm] per liter [1], p < 0.001). Comparison of supine and tilt positions revealed marked changes in heart rate and blood pressure means and variances consistent with more pronounced decreases in intracardiac filling pressures and unloading of the arterial baroreceptors. A further progressive decrease in the vagal modulation of heart rate by lung volume was observed in the 0.12-0.5-Hz band, with a near-linear response of magnitude of respiratory sinus arrhythmia over a range of estimated central venous volume. Transfer function analysis can detect changes in autonomic response to mild degrees of central hypovolemia, which are insufficient to cause changes in mean heart rate or heart rate variance. This represents evidence for modulation of heart rate control by cardiopulmonary baroreceptors. A near-linear relation between magnitude of respiratory sinus arrhythmia and central venous volume suggests that this may have clinical relevance in patient monitoring. PMID- 8428787 TI - Effect of reduced alcohol consumption on blood pressure in untreated hypertensive men. AB - Fifty-four untreated, mildly hypertensive men whose daily alcohol consumption was > or = 28 ml ethanol and who drank at least 4 times per week took part in a randomized, controlled crossover trial. The purpose of the trial was to test the effects of alcohol reduction on blood pressure. After a 2-week familiarization period, the participants were assigned to either a reduced alcohol drinking group or a usual drinking group for 3 weeks (experimental period 1). The situation was then reversed for the next 3 weeks (experimental period 2). The participants were requested to limit their daily alcohol consumption to zero or reduce it as much as possible for the reduced alcohol consumption period. The self-reported alcohol consumption was 56.1 +/- 3.6 (SEM) ml/day during the usual alcohol drinking period and 26.1 +/- 3.0 ml/day during the period of reduced alcohol consumption. Systolic and diastolic blood pressures in the intervention group were found by analysis of variance to be significantly lower (2.6-4.8 and 2.2-3.0 mm Hg, respectively) than those in the control group during experimental period 2 for systolic blood pressure and experimental period 1 for diastolic blood pressure. Significant (3.6 mm Hg) and nonsignificant (1.9 mm Hg) decreases in systolic and diastolic blood pressure, respectively, were observed. The method of Hills and Armitage was used, reducing ethanol in daily alcohol consumption by 28 ml. The lowering effect of reduced alcohol consumption on blood pressure was independent of changes in salt consumption, which were estimated by 24-hour urine collection and body weight.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428788 TI - Enhancement of vasoconstrictor response by a noncalcemic analogue of vitamin D3. AB - To clarify the effects of active vitamin D3 on pressor and vascular responses to vasoconstrictor substances, we studied pressor responses to the intravenous injection of norepinephrine or angiotensin II (Ang II) and vasoconstrictor responses to norepinephrine. Sprague-Dawley rats were given 1,25-dihydroxyvitamin D3 subcutaneously (200 ng/kg per day) for 14 days. The administration of 1,25 dihydroxyvitamin D3 augmented the pressor responses to norepinephrine and Ang II in conscious rats and was associated with a significant increase in serum calcium concentration (11.0 +/- 0.2 mg/dl). To further clarify whether the increased pressor response to vasoconstrictors may be due to the calcemic or direct action of active vitamin D3, we studied the effect of its noncalcemic analogue, 22 oxacalcitriol, and its inactive analogue, 24,25-dihydroxyvitamin D3, on the pressor response to vasoconstrictors in rats. The pressor responses to norepinephrine and Ang II were apparently augmented in 22-oxacalcitriol-treated rats similarly to 1,25-dihydroxyvitamin D3-treated rats. In contrast, the pressor responses were not affected by either 24,25-dihydroxyvitamin D3 or the intravenous infusion of calcium chloride. In an ex vivo experiment using a mesenteric preparation, the vascular sensitivity to norepinephrine was moderately augmented in rats treated with both 22-oxacalcitriol and 1,25-dihydroxyvitamin D3 but was not affected in rats treated with 24,25-dihydroxyvitamin D3. The results suggest that the enhanced pressor responses to norepinephrine and Ang II could be attributed to the direct effect of active vitamin D3 on vasculature rather than to hypercalcemia. PMID- 8428789 TI - Comparative cytogenetic and DNA flow cytometric analysis of 150 bone and soft tissue tumors. AB - Samples from 48 benign and 102 malignant bone and soft-tissue tumors were analyzed cytogenetically and by DNA flow cytometry. Clonal chromosome abnormalities were found in 82 tumors and normal karyotypes in 68; 61 tumors were DNA-non-diploid and 89 were diploid. The cytogenetically abnormal tumors were used for comparison between the 2 types of investigation; 45 of these tumors were DNA-diploid and 37 were DNA-non-diploid. There was, with few exceptions, good correspondence between the quantitative estimates of genomic changes by the 2 methods, indicating that the cells cytogenetically analyzed from short-term cultures are representative of the in vivo cell populations. Discrepancies were primarily found in cases with indexes above 1.5, in which the DNA index was higher than the chromosome index. The chromosome analysis suggested that skewed stemline (G0/G1) peaks in the diploid region in DNA histograms indicate the presence of cell populations with small net quantitative genomic changes, although not all such populations were detected by DNA flow cytometric analysis. The view that one of the peaks in bimodal stemline DNA histograms with narrow peaks represents a non-diploid cell population was also corroborated. On average, the cell populations giving rise to double stemlines in DNA histograms showed quantitatively larger genomic changes than those that gave rise to broad or skewed diploid G0/G1 peaks. The findings indicate that these histogram profiles are not artifactual but reflect chromosomal changes in the tumor parenchyma. PMID- 8428790 TI - Over-expression of p53 nuclear oncoprotein in transitional-cell bladder cancer and its prognostic value. AB - Two hundred and twelve archival bladder-cancer biopsy specimens were analyzed immunohistochemically analysis were correlated to established histological and quantitative prognostic factors and survival of patients during a mean follow-up period of more than 10 years. Twenty-nine percent of tumours were positive for p53 protein, and over-expression was associated with high histological grade, non papillary growth architecture, dense inflammatory cell reaction, DNA aneuploidy, high S-phase fraction, high mitotic frequency and high SD of nuclear area. Progression in T, N and M categories was significantly related to over-expression of p53 protein. In univariate survival analysis, over-expression of p53 predicted poor outcome in the entire cohort, in papillary tumours and in muscle-invasive tumours but not in superficial tumours. In a multivariate survival analysis, over expression of p53 oncoprotein had no independent prognostic value over clinical stage and mitotic index. The results confirm that p53 is involved in the growth regulation of bladder cancer and is certainly a subject for detailed analysis of specific mutations. PMID- 8428791 TI - International variations in the incidence of childhood bone tumours. AB - Bone cancers comprise about 5% of childhood neoplasms. Osteosarcoma, the most common sub-type, shows a somewhat irregular geographic pattern of incidence, with low rates in some Asian (Indian, Japanese, Chinese) and Latin American populations. Incidence is similar in the sexes and rises steeply with age, accompanied by an increasing proportion of tumours localized in the long bones of the legs. Rates in the USA are higher in blacks than in whites, as a result of a higher incidence at ages 10 to 14 and of tumours of the leg bones. The descriptive epidemiology is consistent with early observations linking risk to the amount of bone growth. Ewing's sarcoma is rare in black populations (USA and Africa) and in eastern Asia. Compared with osteosarcoma, a lower percentage of tumours is localized to the long bones, and incidence rises less steeply with age and is accompanied by an increasing proportion of pelvic tumours. Chondrosarcoma is a rare cancer in children (less than 5% of bone cancers), with an age distribution similar to that of osteosarcoma and a sub-site distribution resembling that of Ewing's sarcoma. Little is known of the aetiology of these tumours; there is clearly a strong genetic predisposition in Ewing's sarcoma but, although the proportion of osteosarcoma cases of genetic origin seems to be small, environmental determinants so far suspected can account for only a small fraction of the total cases. PMID- 8428792 TI - Isolation and characterization of an oestrogen-responsive breast-cancer cell line, EFF-3. AB - A novel oestrogen-responsive breast-tumour cell line, EFF-3, has been established from a pleural exudate of a patient with metastatic breast cancer. The cells show morphological and immunohistochemical features consistent with their origin from a metastatic breast carcinoma. The cells aggregate and form sheets in culture, and electron microscopy confirms the presence of cell-surface microvilli and intercellular tight junctions. The epithelial origin of EFF-3 cells was confirmed by their expression of low-molecular-weight cytokeratins and carcinoembryonic antigen. The karyotype of the cells is markedly abnormal and there are large numbers of structurally abnormal chromosomes. EFF-3 cells express oestrogen receptor, oestrogen-receptor mRNA, their growth is oestrogen-responsive, and specific genes are regulated by oestrogens. The pNR-2/pS2 and pNR-25 oestrogen regulated mRNAs are induced 15- and 13-fold respectively by oestrogen, whereas the oestrogen-receptor and cathepsin D mRNAs are not regulated. This pattern of regulation differs from that reported previously for other cell lines. The EFF-3 cell line should be useful for studying the mechanisms involved in oestrogen stimulated proliferation and the factors determining the regulation of specific genes by oestrogens. PMID- 8428793 TI - Differential regulation of gelatinase B and tissue-type plasminogen activator expression in human Bowes melanoma cells. AB - A comparison of the production of tissue-type plasminogen activator (t-PA) and gelatinases A and B was made at the mRNA and protein levels in human Bowes melanoma cells treated with phorbol myristate acetate (PMA). Immunocytochemical analysis confirmed previous quantitative data on PMA-mediated induction of t-PA. It also showed that t-PA immunoreactivity can be restrained to the local environment of the producing cell, most probably by interaction with extracellular matrix components. Zymographical analysis showed that gelatinase B protein was induced by PMA, whereas gelatinase A remained at the constitutive level. Protein kinase C (PKC) appeared to be involved in this regulation since, after PMA treatment (1) the PKC activity was found to be translocated from the cytosol to the particulate fraction of the cells and (2) addition of staurosporine and H-7 blocked the gelatinase B increase. Northern-blot hybridization showed a transient rise in t-PA and gelatinase B mRNA levels whereas gelatinase A mRNA levels remained unchanged. When c-fos and c-jun mRNAs were investigated, only that of c-fos was affected by PMA. Activation by PMA can be kinetically ordered as follows: translocation of PKC to the membrane fraction, transcription of the c-fos gene and eclipsing of gelatinase B mRNA, increase in steady-state mRNA levels of t-PA and gelatinase B and, finally, secretion of t-PA and gelatinase B glycoproteins. Our data also suggest that various proteases that are known to cooperate in the remodeling of the extracellular matrix can be differently regulated in one tumor-cell type. PMID- 8428794 TI - Analysis of the relationship between stage of differentiation and NK/LAK susceptibility of colon carcinoma cells. AB - NK and LAK cells which are able to lyse tumor target cells in an MHC-unrestricted manner are not equally effective against targets of the same nature. In the case of colorectal tumors, some cells are highly sensitive, whereas others are resistant to NK and can even be quite resistant to LAK-mediated lysis. In the present paper, we tried to correlate the stage of differentiation of 17 human colorectal tumor cell lines with their NK- or LAK-cell susceptibility. It was observed that NK cells killed colorectal target cells independently from their stage of differentiation defined according to histopathological criteria from xenografting in nude mice. NK susceptibility was not correlated either with in vitro-defined criteria of differentiation, such as cell polarity and morphology, brush-border enzyme expression and CEA production. A LAK-resistant HT-29 sub-line (HT-29 LAK) was selected which could not be distinguished from HT-29 in terms of features of differentiation. It was further observed that HT-29 Glc-/+ cell line, a highly differentiated enterocytic-like variant of HT-29, obtained after glucose starvation, was killed by LAK cells as efficiently as the moderately differentiated parental HT-29, and that Caco-2 cells, which differentiate spontaneously after confluence in standard culture conditions, were equally sensitive to NK-mediated lysis whatever their stage of differentiation. In contrast, HT29 MTX10(-5), a highly differentiated mucus-secreting variant of HT29 obtained by methotrexate selection, was much more resistant to LAK cells than parental HT29 cells. PMID- 8428795 TI - Structure-activity relationships of four anti-cancer alkylphosphocholine derivatives in vitro and in vivo. AB - This study was carried out to investigate whether structure-activity relationships of alkylphosphocholines, a new group of anti-neoplastic agents, which had been detected in methylnitrosourea(MNU)-induced rat mammary carcinoma, can be transferred to in vitro systems. Therefore, the anti-neoplastic activity of 4 alkylphosphocholines (APCs) was compared in 6 tumor cell lines in vitro and in MNU-induced rat mammary carcinoma in vivo. The in vitro system consisted of 2 rat mammary-carcinoma-derived cell lines (1/C2 and 1/C32), as well as 2 human mammary-gland (MDA-MB-231 and MCF-7)- and gastrointestinal tract (HT-29 and KB) derived tumor cell lines. As assessed by both cell counting and MTT-assay, the ranking of concentrations effecting 50% growth inhibition (IC50) was parallel in all cell lines for octadecylphosphocholine (18:0-PC), octadecenyl-(trans-9.10) phosphocholine (t-18:1-PC) and octadecenyl-(cis-9.10)-phosphocholine (c-18:1-PC). Only hexadecylphosphocholine (16:0-PC) differed in its activity, being least active in 1/C2, 1/C32 and MDA-MB-231 cells, moderately active in KB and MCF-7 cells, and most active in HT-29 cells. The IC50 concentrations of APCs in the 2 rat mammary carcinoma cell lines significantly correlated with dosages effecting a 50% tumor growth delay in vivo. Remarkably, the 2 gastrointestinal cell lines were more sensitive to APC exposure than the mammary-carcinoma cell lines. In all cell lines except KB cells, growth-stimulation effects were seen in the concentration range preceding the anti-proliferative activity; in vivo, however, no accelerated cancer growth was observed. The in vitro system failed to describe the superior therapeutic ratio of c-18:1-PC, as assessed in vivo, because it does not take the relative sensitivity of tumor vs. normal cells into account. Complementary in vivo trials are therefore indispensable for a final evaluation. Comparison of the 2 in vitro assays shows good agreement of the interrelationship of IC50 values, those obtained by MTT assay being on average 25% higher than those obtained from cell counting. PMID- 8428796 TI - Monocyte-conditioned media possess a novel factor which increases motility of cancer cells. AB - A number of cytokines have been reported to increase the cell motility (scatter) of cohesive cell colonies. We report here a novel scattering factor produced by human monocytes. Media from both stimulated and non-stimulated human monocytes or the monocytic cell line, U937, increased the cell motility of 2 human colon cancer cell lines, HT115 and HT29, but not the canine epithelial cell line MDCK. Motility was assayed by cell dissociation from carrier beads and colony scattering. HT115 cells were strongly scattered by the conditioned media but not by interleukins IL-1, 2, 4, 6, 8, 10, TNF alpha, TGF beta, EGF, GM-CSF, IGF-I, PDGF, interferon-gamma. This factor is distinct from hepatocyte growth factor/scatter factor (HGF/SF), since the activity was not blocked by anti-HGF/SF antibody. The activity was reduced by treatment with acid, heat, trypsin and dithiothreitol; this, together with gel filtration, suggests that the factor is a protein (MW 40 to 60 kDa). This new factor, which is secreted from monocytes and the monocytic cell line, U937, has the ability to increase the motility of cancer cells and may be important in controlling the behaviour of tumours in vivo. PMID- 8428797 TI - Combination effect of vaccination with IL2 and IL4 cDNA transfected cells on the induction of a therapeutic immune response against Lewis lung carcinoma cells. AB - In order to develop a more effective method of immunotherapy we have transfected mouse interleukin-2 (IL2) or mouse interleukin-4 (IL4) cDNA into a spontaneous non-immunogenic murine lung cancer. Lewis lung carcinoma (LLC). IL2 cDNA transfection more strongly decreases tumorigenicity of LLC than IL4 cDNA transfection. Recombinant-human-IL2 treatment of mice that were transplanted with untransfected LLC could not prolong their survival. In contrast, vaccination with IL2-cDNA-transfected LLC (LLC-IL2) and LLC-IL2 mixed with IL4-cDNA-transfected LLC (LLC-IL4) could significantly suppress tumor growth of LLC in a tumor specific manner. The vaccination with LLC-IL2 mixed with the same number of LLC IL4 cells was more suppressive to the growth of LLC than that with LLC-IL2 cells alone, while LLC-IL4 vaccination alone was ineffective. Nude, severe-combined immune-deficient (SCID) and beige mice were unable to reject LLC-IL2 cells. However, immunodeficient mice responded to LLC-IL2, but not to LLC, since their survival times after transplantation with LLC-IL2 cells were significantly longer than the survival time of normal or immunodeficient mice transplanted with untransfected LLC cells. We conclude that vaccination with IL2-producing tumors and, with more pronounced effect, in combination with IL4-producing tumors, is able to induce an immune response to this normally non-immunogenic tumor. Tumor rejection appears to be achieved by the combined activity of CTL and NK cells. This strategy has potential for new immunotherapeutic interventions in cancer patients. PMID- 8428798 TI - Immortalization of normal human fibroblasts by treatment with 4-nitroquinoline 1 oxide. AB - Normal human fibroblasts (the OUMS-24 strain), derived from a 6-week-old human embryo, were transformed (into the OUMS-24F line) and immortalized by repeated treatments (59 times) with 4-nitroquinoline 1-oxide (4NQO). Treatment began during primary culture and ended at the 51st population doubling level (PDL). At the 57th PDL (146 days after the last treatment), morphologically altered, epithelial-type cells appeared, began to grow and became immortal (now past the 100th PDL). However, the control fibroblasts, which were not treated with 4NQO, senesced at the 62nd PDL. The finding that extensive, repeated treatments with 4NQO are required for the immortalization of normal human cells, indicates that multiple mutational events are involved in the immortalization of human cells in general. In other words, immortalization itself seems to be a multi-step process. Karyotypic analysis showed that many cells were hypodiploid before immortalization, but that afterwards chromosomes were distributed broadly in the diploid to tetraploid regions. The immortalized cells showed amplification and enhanced expression of c-myc. Two-dimensional electrophoretic analysis showed that the number of disappearing cellular proteins was greater than the number of the newly appearing ones after the cells became immortalized. Since the immortalized cells showed neither anchorage-independent growth nor tumorigenicity, they are useful for studying factors that can contribute to multi step carcinogenesis in human cells. In addition, genetically matched normal (OUMS 24) and immortalized (OUMS-24F) cells will be useful for analyzing the genes related to cellular mortality and immortalization. PMID- 8428799 TI - Phenotypic and molecular analysis of Ph1-chromosome-positive acute lymphoblastic leukemia cell lines. AB - We have established 2 Philadelphia chromosome (Ph1)-positive acute lymphoblastic leukemia (ALL) cell lines, designated PALL-1 and PALL-2, from distinct adult Ph1 positive ALL patients. PALL-1 was established in nude mice, and PALL-2 was established in culture. Both retained the Ph1 chromosome and expressed the ALL type bcr/abl chimeric mRNA containing the junction of the first exon of BCR gene (e1) and second exon of c-abl gene (a2). PALL-1 and PALL-2 expressed CD34 surface antigen which is characteristic of early hematopoietic progenitor cells. PALL-2 expressed antigens for both pre-B and early myeloid cells and had rearrangements of both the heavy chain of immunoglobulin gene and the beta chain of T-cell receptor gene. Both PALL-1 and PALL-2 expressed detectable levels of p53 gene RNA. Polymerase-chain-reaction-single-strand conformation polymorphism (PCR-SSCP) analysis of the p53 gene showed a normal pattern of mobility in both cell lines. Taken together, the 2 cell lines had features of Ph1-positive ALL: (i) hematopoietic progenitor cells with pre-B-cell phenotype and, (ii) activation of e1-a2 type bcr/abl oncogene without alterations of p53 gene. These unique lines should provide a valuable tool for studying the pathogenesis of Ph1-positive ALL. PMID- 8428800 TI - Anti-metastatic vaccination of tumor-bearing mice with IL-2-gene-inserted tumor cells. AB - IL-2 gene was introduced through retroviral vectors into the highly malignant and poorly immunogenic 3LL-D122 clone. Both high and low D122-IL-2 secretors showed elimination of tumorigenicity in syngeneic immune-competent mice; however, in nude mice only the high IL-2 secretor showed reduced tumorigenicity as compared with parental D122 cells. Also, following intravenous inoculation, only the high IL-2 secretor showed reduced generation of metastases, whereas the low IL-2 secretors were as highly metastatic as parental cells. These results seem to indicate that low levels of IL-2 secreted by tumor cells are sufficient to activate T cells, while higher levels are needed in order to activate non-T-cell effectors. D122-IL-2 secretors induced high levels of anti-tumor CTL, while their sensitivity to the lytic activity of these CTL was similar to the sensitivity of D122 cells, thus indicating that the elevation of immunogenicity of IL-2 secretors was essentially due to the secreted IL-2. In accordance with CTL induction, pre-immunization with IL-2 secretors protected mice against subsequent challenge of parental cells. Moreover, immunization in an "immunotherapy protocol" i.e., vaccination of mice carrying an established tumor of parental D122 cells with inactivated D122-IL2 infectants, almost completely eliminated the generation of lung metastases by D122 cells, thus providing a rationale for the use of IL-2 gene transferred tumor cells as a modality for treatment of metastasis. PMID- 8428801 TI - Persistence of plasmin-mediated pro-urokinase activation on the surface of human monocytoid leukemia cells in vitro. AB - Human leukemia cell lines, unlike those from adherent tumors, have been shown to continuously activate the pro-urokinase (pro-u-PA) they produce. In the present study we found that, in normal cell-culture conditions in 10% FCS the plasminogen activation cascade works continuously on monocytoid leukemia cells, which expressed plasmin activity and active u-PA on their cell surface. This plasmin catalyzed the conversion of the produced pro-u-PA to active 2-chain urokinase (tcu-PA), and was derived from bovine serum plasminogen by the activity of cell bound tcu-PA. Plasmin generation was abolished and pro-u-PA accumulated in cell cultures that were grown for several days, either in the presence of serum thoroughly depleted of plasminogen, or in the presence of 1 mM tranexamic acid. Plasmin generated on the cell surface was found to be present in 2 enzymatically active fragments, of M(r) 85,000 and M(r) 50,000, which were slowly released into the growth medium. These fragments could activate pro-u-PA in serum-free growth medium. Most of the bound plasmin could be washed off cells with 10 mM tranexamic acid, but complete removal of plasmin from the cell surface required washing of the cells with acid-glycine pH 3.0. PMID- 8428802 TI - Malaria: the threat grows. PMID- 8428803 TI - Perceived vulnerability to HIV/AIDS in the US and Zimbabwe. AB - The majority of nurses in the US and Zimbabwe feel they have sufficient knowledge to protect themselves from acquiring AIDS, according to a study conducted by the authors, leading them to recommend that nursing students be allowed to openly discuss their feelings about vulnerability to contracting AIDS in the workplace. PMID- 8428804 TI - Breaking the silence: the need for counselling of HIV/AIDS patients. AB - The care and management of persons with HIV infection and AIDS pose tremendous challenges to families, as individuals with the disease will more likely be taken care of within their homes when discharged from hospital. For effective care to be provided, relatives and families must be able to provide emotional, psychological and socioeconomic support. The capacity of the relatives to cope with the burden of an individual with HIV/AIDS--a terminal illness often associated with stigma in many communities--may be a major determinant for acceptance or rejection of the HIV/AIDS individual. Below, a report on a study to identify the possible factors that influence the capacity of relatives and family to cope with the needs of the patient with HIV/AIDS in an urban Zambian community. PMID- 8428805 TI - Health services for refugees in countries of second asylum. AB - As successive groups of refugees reach countries of second asylum, refugee health care must be reinvented for each new group. But how can we bridge the one-to-two year lag time between resettlement and publication of studies of specific cultures and thus render effective health services for refugees from the time of resettlement? Below, health problems common to refugees in countries of second asylum are identified and a community-based system for addressing their healthcare needs is proposed. The nursing process and principles of community health nursing are key concepts. PMID- 8428806 TI - Administration of patient care: theoretical aspects. AB - In Brazilian hospitals, nursing administrations place priority on the interests of the hospital and doctors. But according to the authors the focus should centre more on the patient. Below, they present their case based on behavioural theory. PMID- 8428807 TI - Should the nurse be blamed? PMID- 8428808 TI - Women, health and development. AB - Ways to improve the health of women were discussed at the technical discussions at the World Health Assembly in May 1992. Nurses are being particularly urged to join up with the women's pressure groups in efforts to sensitize the media and politicians. ICN, for its part, will be enlisting nurses' cooperation in its 1994 International Nurses' Day campaign, which will show nurses how to educate women to improve their health, and thus that of their children and families. The WHA discussions produced two important recommendations: the establishment of indicators to monitor the evolution of women's health and of a Commission on Women's Health that would work toward health rights for women. Below, a summary of the discussions and proposals for action. PMID- 8428809 TI - AIDS: a community challenge. PMID- 8428810 TI - A national study of the efficiency of hospitals in urban markets. AB - Using a sample of 3,000 urban hospitals, this article examines the contributions of selected hospital characteristics to variations in hospital technical efficiencies, while it accounts for multiple products and inputs, and controls for local environmental variations. Four hospital characteristics are examined: hospital size, membership in a multihospital system, ownership, and payer mix (managed care contracts, percent Medicare, and percent Medicaid). Ownership and percent Medicare are consistently found to be related significantly to hospital efficiency. Within the ownership variable, government hospitals tend to be more efficient and for-profit hospitals less efficient than other hospitals. Higher percentages of Medicare payment are negatively related to efficiency. While not consistently significant across all five of the MSA size categories in which the analyses are conducted, possession of managed care contracts, membership in a multihospital system, and size all are consistently related positively to hospital technical efficiency. These variables are also all significant when the hospitals are examined in a combined analysis. Percent Medicaid was not significant in any of the analyses. Implications for policy and the need for methodological work are discussed. PMID- 8428811 TI - The financial performance of diversified hospital subsidiaries. AB - Despite its proliferation, we know relatively little about the impact of hospital restructuring to offer new services. This exploratory study examines the relationship between types of services offered and financial performance among separately incorporated subsidiaries of acute care hospitals. We draw data from the subsidiaries of all hospital firms operating in one state (Virginia) that requires reporting by all such firms. Results from multiple regression analyses of 1987 data indicate that units that existed longer, produced health care or related products, or were nonprofit subsidiaries of nonprofit firms tended to be more profitable than the other subsidiaries. PMID- 8428812 TI - The relationship between adjusted hospital mortality and the results of peer review. AB - This study assessed the relationship between the Health Care Financing Administration adjusted mortality rate for a hospital and the errors in care found by the peer review process. The three data sets used were: (1) the 1987 1988 completed reviews from 38 peer review organizations (PROs) of 4,132 hospitals and 2,035,128 patients; (2) all 1987 hospital mortality rates for Medicare patients as adjusted by HCFA for patient mix; and (3) the 1986 American Hospital Association Survey. The PRO data were used to compute the percentage of cases reviewed from each hospital confirmed by a reviewing physician to have a quality problem. The average percentage of confirmed problems was 3.73 percent with state rates ranging from 0.03 percent to 38.5 percent. The average within state correlation between the problem rate and the adjusted mortality rate for all PROs was .19 (p < .0001), but the correlations were much higher for relatively homogeneous groups of hospitals, .42 for public hospitals and .36 for hospitals in large metropolitan statistical areas (MSAs). These results suggest that the HCFA adjusted hospital mortality rate and the PRO-confirmed problem rate are related methods to compare hospitals on the basis of quality of care. Both methods may compare quality better if used within a group of homogenous hospitals. PMID- 8428813 TI - The effect of HMOs on premiums in employment-based health plans. AB - This study documents the effect of HMOs on premiums in employment-based health plans. We analyzed a survey of Minnesota employers conducted in 1986. Among 922 usable observations, 239 firms offered HMOs in addition to fee-for-service (FFS) health plans. We estimated an equation for the probability of offering an HMO, followed by equations for HMO enrollment share, and HMO and FFS premiums. The weighted average HMO and FFS premium in firms that offer HMOs was compared to the premium of FFS-only firms. We found that offering an HMO raises the average premium for family coverage health insurance by $25.14 per month and for single coverage by $3.68 per month. This effect was smaller for firms in the Twin Cities metropolitan area. HMOs may be viewed as a progressive and innovative health care benefit, but they are likely to increase firms' health insurance premiums. PMID- 8428814 TI - Insurance incentives for ambulatory surgery. AB - This study is an attempt to address both the extent to which surgical procedures on an outpatient basis substitute cost-effectively for inpatient procedures, and whether or not an insurance policy's financial incentives increase the volume of outpatient surgical procedures. In particular, given an insurance product of a given composition: What is the probability that the insured will have surgery? and if a surgery does take place, what is the probability that it will occur in an outpatient setting? Finally, the article assesses the implication of such products on the total cost of care by quantifying the insurance plans along two parameters, the relative user price for outpatient versus inpatient surgery and the absolute price for the inpatient surgery. The results indicate that insurance policies that offer relatively lower out-of-pocket payments for ambulatory surgery do not increase the probability that surgery will be done in the ambulatory setting. However, higher out-of-pocket payments for surgery, regardless of site, do reduce the surgery rate. There are other patient and market characteristics, especially the availability of freestanding surgery firms, that do influence the location of surgery. PMID- 8428815 TI - Relationship of physician ratings of severity of illness and difficulty of clinical management to length of stay. AB - In a concurrent prospective study, medical and surgical residents rated the severity of illness and difficulty of clinical management of each of their patients within 24 hours of admission, and on a daily basis throughout the patient's stay. Data were collected on consecutive admissions resulting in 661 admissions with complete data for analysis. Results indicate that difficulty and severity are correlated, each explaining variations in length of stay (LOS), and together explaining up to 44 percent. Four alternative measures are tested, first day values, average values over the stay, peak or highest values, and a zero-one measure indicating whether or not the severity or difficulty fluctuated over the stay. First-day and average measures of severity and difficulty explain little variation in LOS; peak and fluctuating measures are highly explanatory. After adjusting for diagnosis-related groups (DRGs), fluctuating severity adds 34 percent, and adjusting for both DRGs and severity, fluctuating difficulty adds 10 percent for a total of 53 percent variance explained. In comparable results, peak severity adds 21 percent, and peak difficulty 4 percent, for a total of 34 percent variance explained. Findings indicate that difficulty had independent value as a predictor, and the high explanatory power of the fluctuating measures suggests that a third dimension, instability, may be as important as severity and difficulty in explaining LOS. PMID- 8428816 TI - Tear analysis. PMID- 8428817 TI - Self-mutilative behavior in horses. PMID- 8428819 TI - Welfare position on disabled livestock approved. PMID- 8428818 TI - Forum urges commitment to resolving overpopulation. PMID- 8428820 TI - Shedd Aquarium, Boehm recuperate from beluga deaths. PMID- 8428821 TI - Views on extra-label use expounded at symposium. PMID- 8428822 TI - Compendium of animal rabies control, 1993. National Association of State Public Health Veterinarians, Inc. PMID- 8428823 TI - Rabies epizootic continues to grow. PMID- 8428824 TI - 1991 professional incomes of US veterinarians. PMID- 8428825 TI - Graduate studies for the veterinary clinician. PMID- 8428826 TI - Let's look like professionals. PMID- 8428827 TI - Impact of expenditures for veterinary services and medical supplies on dairy farm productivity and profitability. AB - The value of the marginal product (VMP) for veterinary services and medical supplies (VETMED), and the profit maximizing level of VETMED were estimated for dairy producers. Data from the Pennsylvania Farmers Association-Dairy Farm Business Analysis system during the years of 1986 to 1990 were used to evaluate the functional relationship between production and expenditures for VETMED. Other input variables examined were man-year equivalents of labor, asset values, value of feed fed, and culling rate. Data were screened to reflect economically viable dairy farms in Pennsylvania, and 173 such farms participated for each of the 5 years analyzed. The VMP was estimated for 1990. Profit maximizing levels for VETMED were estimated for 1990 holding other input variables at their mean values. Mean expenditures for VETMED were $2,606/farm, or $43/cow in 1990. The VMP for VETMED was estimated to be $3.22 or $4.98, depending on the method of calculation. In other words, the marginal dollar spent on VETMED generated $3.22 ($4.98) in additional revenue from milk production. The profit maximizing level of expenditures for VETMED was $138/cow, substantially more than the mean, indicating the potential for farms in this data set to improve profitability through additional expenditures on VETMED. PMID- 8428828 TI - Issues deserving renewed attention in 1993. PMID- 8428829 TI - 1993 Report of the AVMA Panel on Euthanasia. PMID- 8428830 TI - Use of synthetic prostaglandin E1 (misoprostol) for prevention of aspirin-induced gastroduodenal ulceration in arthritic dogs. AB - A randomized, double-blind, controlled study was performed with 18 arthritic dogs administered aspirin (25 mg/kg of body weight, PO, q 8 h) and excipient (control group) or aspirin and misoprostol (100 micrograms, PO, q 8 h). Dogs in the misoprostol (n = 10) and control (n = 8) groups were primarily compared by use of sequential gastroduodenoscopy, changes in PCV, and prevalence of clinical signs of gastrointestinal disturbance over a 14-day treatment period. The misoprostol/aspirin-treated group had significantly (P < 0.05) less gastroduodenal hemorrhage and ulceration and a significantly (P < 0.05) lower prevalence of vomiting than did the control group. PMID- 8428831 TI - Evaluation of a method of ureteroneocystostomy in cats. AB - Renal transplantation is a successful treatment for terminal renal failure in cats. Ureteral obstruction at the bladder wall or stoma has been a technical complication encountered in approximately 50% of clinical transplants. The small (0.4 mm) diameter of the feline ureter makes standard techniques described for ureteroneocystostomy unsatisfactory for cats. In this study, we used microsurgical techniques to oppose ureteral mucosa to intact bladder mucosa in an attempt to form a stricture-free stoma. In each of 5 anatomically normal cats, 1 ureter was microsurgically implanted in the bladder. Ultrasonographic examination of the kidneys, ureters, and bladder was performed twice weekly for 8 weeks. Excretory urography was performed at postoperative weeks 1, 2, 4, 8, and 12. Biopsy specimens were taken from the kidney on the surgically treated side 4 weeks after surgery. At 12 weeks, the kidney, ureter, and ureterovesical junction from the treated side were removed and submitted for histologic evaluation. At 1 week, all cats had enlargement of the kidney, renal pelvis, and ureteral lumen on the treated side. This enlargement gradually decreased, and by week 8, there was no difference in comparison with the control side. Ureteroneocystostomy that requires tunneling of the ureter through the bladder wall may result in hydroureter and hydronephrosis, which may resolve. Recognition of these changes may prevent unwarranted surgical intervention in cases of suspected obstruction. The technique described in this study has been used in 15 cats receiving renal allografts. None required surgical correction of ureteral obstruction. PMID- 8428832 TI - Intratumoral chemotherapy with cisplatin in oily emulsion in horses. AB - Twenty horses with 30 lesions were studied to evaluate the effects of intratumoral chemotherapy with cisplatin in sesame oil on equine sarcoids (n = 19), squamous cell carcinomas (n = 7), and squamous cell papillomas (n = 4). Treatment consisted of 4 sessions of intratumoral cisplatin chemotherapy at 2 week intervals. A controlled-release formulation of cisplatin in sesame oil was used to limit drug egress from the injection site. Mean dosage per session was 0.97 (+/- 0.17, SEM) mg of cisplatin/cm3 of tumor tissue treated for tumor volumes ranging from 10 to 20 cm3. Dosage tended to be slightly higher for smaller tumors and slightly lower for larger tumors. Tumor regression was observed in all horses. Complete response was observed in 18 of the sarcoids, 5 of the squamous cell carcinomas, and 4 of the squamous cell papillomas. The mean relapse-free interval was 21.6 and 14 months in horses with sarcoid and carcinoma/papilloma, respectively. The 1-year relapse-free rates were 87 and 65% for equine sarcoid and carcinoma/papilloma, respectively. In horses with relapse, 70% had tumor recurrence outside the treated field. Cisplatin-related local toxicosis was minimal. Intratumoral cisplatin chemotherapy was found to be a practical and effective treatment of sarcoid and squamous cell carcinoma/papilloma in horses. PMID- 8428833 TI - Prophylactic efficacy of tilmicosin for bovine respiratory tract disease. AB - The prophylactic administration of injectable tilmicosin for pneumonia in weaned beef calves was investigated in 1,806 animals. Comparisons were made among calves receiving an "on-arrival" injection of tilmicosin, calves receiving a single injection of long-acting oxytetracycline, and calves receiving no prophylaxis. Morbidity and mortality attributable to pneumonia, morbidity and mortality attributable to all causes, and case fatality were significantly lower in the group of calves that received tilmicosin, compared with calves that received long acting oxytetracycline and calves that received no prophylactic antibiotic. Mean time to initial pneumonia treatment was significantly extended in calves that received prophylaxis, compared with those that received no antibiotic on arrival at the feedlot. Calves that received tilmicosin gained significantly more weight than calves that received oxytetracycline. Calves that were not treated for pneumonia during the trial period gained significantly more weight than did those calves that were treated for pneumonia regardless of experimental group. The majority of mortalities were attributable to fibrinous pneumonia (31/34). Important bacterial isolates (Pasteurella spp, Haemophilus somnus, Actinomyces pyogenes) obtained at necropsy did not have resistance to tilmicosin in association with administration of tilmicosin as prophylaxis for pneumonia. However, bacterial resistance to trimethoprim/sulfonamide and to oxytetracycline were commonly found in these postmortem isolates. PMID- 8428834 TI - Naltrexone-induced pruritus in a dog with tail-chasing behavior. AB - Naltrexone, an opioid antagonist recently advocated for the control of stereotypic behavioral patterns in horses and dogs, induced an intense dose related pruritic reaction in a dog with tail-chasing and self-mutilating behavior. Used in human beings primarily as an adjunct in the treatment of opioid abusers, naltrexone's adverse effects include dermatologic signs, such as alopecia and pruritus, observed in < 1% of human patients. In contrast to previously reported findings, naltrexone treatment was of limited benefit to the dog of this report. During administration of the drug, stereotypic patterns were reduced but did not disappear. Although naltrexone may be effective in controlling certain obsessive-compulsive disorders, its inconsistent therapeutic value, prohibitive cost to pet owners, and side effects such as the pruritus observed in the dog of this report may restrict use of this drug. PMID- 8428835 TI - Transvenous coil embolization for treatment of patent ductus venosus in a dog. AB - A congenital intrahepatic portosystemic shunt was successfully closed in a 3 month-old Golden Retriever by use of a transvenous coil embolization procedure. The pup's patent ductus venosus was occluded by placing 8 Dacron fiber-covered, stainless steel, spring embolization coils into the shunt vessel through a catheter placed through the jugular vein. Four separate embolization procedures were performed to gradually close the portosystemic shunt and to allow intrahepatic portal perfusion to reform. Transvenous embolization may prove to be a better therapeutic alternative than surgery for correction of intrahepatic shunts because of its decreased invasiveness, lower mortality, and ability to gradually narrow the shunt lumen. PMID- 8428836 TI - Cor triatriatum dexter in two dogs. AB - Cor triatriatum dexter is a congenital heart defect in which the embryologic right sinus venosus valve persists as a septum within the right atrium. Cor triatriatum dexter was diagnosed in 2 dogs on the basis of clinical signs, two dimensional echocardiography, and cardiac catheterization. In 1 of the dogs, the condition was successfully treated by surgical resection of the intra-atrial septum. In the second dog, the defect was associated with an incomplete persistent cranial left vena cava and Ebstein's anomaly; surgery was declined. PMID- 8428837 TI - Preoperative diagnosis of tumors of the brachial plexus by use of computed tomography in three dogs. AB - Three dogs with forelimb lameness of 3 months' to 1 year's duration were examined by computed tomography and determined to have a tumor of the brachial plexus. In each case, the clinician had been unable to determine the cause of lameness by other means, and in 2 dogs, surgery had been performed on the affected limb for unrelated conditions prior to diagnosis of the tumor. Computed tomography was performed by use of a third-generation scanner, with dogs under general anesthesia and positioned in dorsal recumbency. Intravenous contrast enhancement with iodinated contrast material was used to help differentiate vascular structures, and a 5-mm scanning width allowed detection of small tumors. In all dogs, approximate tumor location in the transverse plane, invasiveness, and relationship to surrounding structures compared favorably between computed tomographic images and surgical findings. PMID- 8428838 TI - Chronic vaginal prolapse during pregnancy in a bitch. AB - A 4-year-old bitch was referred with chronic vaginal prolapse. It was first noticed during estrus, about 58 days prior to referral. Artificial insemination (AI) was performed twice after reducing the prolapse manually. Abdominal radiography (53 to 55 days after AI) was performed by the attending veterinarian and the bitch was determined to be not pregnant. Serum progesterone and estradiol concentrations were 1.38 ng/ml and 1.79 pg/ml, respectively. With the bitch under general anesthesia, the vaginal prolapse was partially reduced and a hysteropexy was performed. Six days after surgery, the bitch delivered a live male pup. It is recommended that bitches so affected should not be bred, and that those not required for breeding should be ovariectomized. PMID- 8428839 TI - Dermoid cyst in a bull. AB - A 3-year-old bull was examined because of a fluctuant mass on the ventral midline of the cranial portion of the cervical area. The mass originally developed after the bull was handled in a squeeze chute for routine health care. The mass was aspirated and incised repeatedly during the 18 months before referral. The mass was excised at the referral center, and the histologic diagnosis was dermoid cyst. The mass had not redeveloped by 18 months after surgery. Dermoid cysts are most often seen in Rhodesian Ridgeback dogs, although a similar condition has been reported in a 5-week-old calf. PMID- 8428840 TI - Nasopharyngeal grass foreign body in eight cats. AB - Nasopharyngeal grass foreign bodies were diagnosed in 8 indoor/outdoor cats. Each cat was anesthetized with xylazine/ketamine, and the foreign bodies were extracted from the nasopharynx by use of mosquito forceps. PMID- 8428841 TI - Choledocholithiasis attributable to a foreign body in a horse. AB - Cholelithiasis is the most common cause of biliary obstruction in horses. Proposed mechanisms include ascariasis, biliary stasis, ascending biliary infection, and changes in bile composition. In this horse, a foreign body acted as the nidus for bile-salt deposition and ascending cholangitis. Clinical signs (intermittent abdominal pain, icterus, and pyrexia) in conjunction with high serum activity of enzymes indicative of obstructive biliary disease led to a tentative diagnosis of cholelithiasis. Ultrasonography was used to confirm the diagnosis. Postmortem examination revealed a 7-cm wooden stick to be the core of a cholelith found in the common bile duct. PMID- 8428842 TI - Evaluation of single-agent chemotherapy for treatment of clinically evident osteosarcoma metastases in dogs: 45 cases (1987-1991). AB - A study was undertaken to determine the effect chemotherapy had when used to treat 45 dogs with measurable metastatic osteosarcoma. The primary tumor was histologically confirmed as an osteosarcoma in each case. Thirty-nine dogs had the primary tumor surgically removed. Twenty-four of these dogs were treated adjunctively with cisplatin (70 mg/m2 of body surface, IV, q 3 weeks; median 2 doses, range 1 to 6 doses) prior to the onset of metastasis. The remaining 6 dogs from which the primary tumor was not surgically removed were diagnosed as having metastatic osteosarcoma in addition to the primary tumor on initial examination. The median time from initial examination until the development of metastatic disease was 115 days (range, 27 to 1,199 days). The location of the metastatic disease was lungs (31 dogs), bone (3 dogs), soft tissue (1 dog), and multiple sites including lungs, bone, and soft tissue sites (10 dogs). The metastatic lesions were confirmed by pretreatment biopsy (n = 8) or cytologic evaluation (n = 2) in 10 cases and at necropsy in 27 cases. The remaining 8 cases were diagnosed radiographically as multiple metastatic lesions in the lungs consistent with metastatic osteosarcoma. The metastatic disease was treated with cisplatin in 31 dogs (70 mg/m2, IV, q 3 weeks; median 2 doses, range 1 to 4 doses), doxorubicin in 11 dogs (30 mg/m2, IV, q 3 weeks; median 2 doses, range 1 to 3 doses), and mitoxantrone in 3 dogs (5 mg/m2, IV, q 3 weeks; median 2 doses, range 1 to 3 doses).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8428843 TI - Radiographic, epidemiologic, and clinical aspects of simultaneous pleural and peritoneal effusions in dogs and cats: 48 cases (1982-1991). AB - In this study, we found that the rate at which radiographically diagnosed simultaneous pleural and peritoneal effusions (double effusion [DE]) developed was highest in dogs and cats with infectious causes of pleuritis and/or peritonitis and in those with pancreatitis. However, DE were observed more frequently in dogs and cats with neoplastic and cardiovascular diseases. Nonneoplastic liver disease was also documented as a cause of DE in the population of this study. Frequency of DE was increased in males and in animals > 6 years old. The finding of simultaneous pleural and peritoneal effusions can be considered an indicator of disease severity, and warrants a poor to grave prognosis. PMID- 8428844 TI - Lymphocytic/plasmacytic colitis in cats: 14 cases (1985-1990). AB - Lymphocytic/plasmacytic colitis was diagnosed in 14 cats during a 5-year period. Purebred cats were affected significantly (P < 0.001) more often than were nonpurebred cats. Six cats were male and 8 were female. Mean age at onset of clinical signs was 5.1 years (range, 0.5 to 9 years). Hematochezia, observed in 13 cats, was the most commonly reported sign; diarrhea was reported in 11 cats. Mildly high serum alanine transaminase activity and hypokalemia were frequent biochemical abnormalities. Campylobacter sp was cultured from the feces of 1 cat. On endoscopic examination, petechia and hyperemia of the colonic mucosa were detected in 7 of 8 cats. Cats were initially treated with dietary management alone or with a combination of dietary and pharmacologic management. Clinical signs in 7 of 11 treated cats completely resolved, whereas signs in 3 cats were considered improved. One cat was euthanatized when an inadequate response to treatment was observed. Most cats were eventually maintained on dietary management alone. PMID- 8428845 TI - What is your diagnosis? Mineralization in the tendon of insertion of both supraspinatus muscles. PMID- 8428846 TI - Ruminations. PMID- 8428847 TI - Laparoscopic cholecystectomy in pregnancy: contraindicated or indicated? AB - Laparoscopic cholecystectomy has proven to be superior to traditional open cholecystectomy in uncomplicated cases of cholecystitis. The patients experience less pain with less need for narcotics, fewer incisional complications, earlier postoperative ambulation, and a shorter hospitalization. It is our opinion that pregnancy, previously considered an absolute contraindication to laparoscopic cholecystectomy, is now an indication for laparoscopic cholecystectomy in select patients. We describe one of our cases of laparoscopic cholecystectomy during pregnancy. The patient was at 17 weeks gestation and had an unremarkable procedure and a recovery similar to that expected for a non-pregnant patient. Review of current literature has shown that this procedure can be done safely and with an excellent outcome for both the patient and her fetus. PMID- 8428848 TI - Physician/addict: heal thyself or else. High on a pedestal away from reality. PMID- 8428849 TI - Racial minorities at increased risk an Arkansas HIV/AIDS report: autumn 1992. AB - The demographics of the HIV epidemic continue to change. Rates among white homosexuals seem to be leveling off; but there does not appear to be a corresponding decline among black homosexuals. Additionally, having found its first foothold among heterosexual injection drug users, increases in reported infection can be seen among heterosexuals who engage in non-monogamous, unprotected sex. To date, this increase in heterosexual transmission has been more pronounced among black Arkansans than white Arkansans. However, there are definite growth trends among heterosexuals regardless of race. Individuals who engage in risky behavior, specifically injection drug use, unprotected sex, and sex with multiple sex partners, should now consider that the risk of contracting HIV increases daily. Persons who engage in such risky behavior should educate themselves on the transmission of HIV and attempt to minimize their chances of infection. Persons who believe that they may be at risk for HIV should seek immediate testing and treatment. Confidential testing is provided by the Department of Health and individuals may call local offices for times that testing is available. PMID- 8428850 TI - As society struggles with a fault-based malpractice system, responsible parties encourage tort reform-but only with input and support from the medical community. PMID- 8428851 TI - Pharmaceutical research companies testing 92 AIDS medicines. PMID- 8428852 TI - Radiological case of the month. Figure A was leiomyoma and Figure B was a squamous cell esophageal carcinoma. PMID- 8428853 TI - Emotions and asthma. PMID- 8428854 TI - Early wheezing and breast feeding. AB - There is uncertainty as to whether breast feeding protects against subsequent illnesses; it has been suggested that breast feeding may have some protective effects on the severity of long-term outcome of bronchiolitis and in reducing morbidity. We have assessed the effects of breast feeding in 266 patients and 199 controls, all patients were early wheezers (i.e., under 2 years old). Between these groups we found differences in socioeconomic, environmental, and atopic conditions, but there were no significant differences in the numbers who had been breastfed. However, within the group who had had early wheezing we found that infants who had been breastfed for at least one month subsequently had less severe wheezing. These results suggest that breast feeding may be a protective factor for early wheezing only during the first month of life, and a delaying factor in the following months. PMID- 8428855 TI - Is combined therapy of sympathomimetics and theophylline indicated? AB - Bronchodilator effect and toxicity of theophylline 300 mg twice a day (R1), salbutamol 4 mg tid (R2), their combination in higher (200/4mg, R3), and lower doses (100/2mg R4), and placebo (calcium lactate 300 mg) tid (R5) were compared in 25 patients with bronchial asthma in a randomized crossover trial. Statistically significant improvement in forced expiratory volume in one second (FEV1) was observed in all the active treatment groups (R1 to R4) compared with placebo (R5). The mean improvement in FEV1 was 29.0%, 22.0%, 28.0%, 30.0%, and 0.73% in regimen R1, R2, R3, R4, and R5, respectively day 1, and corresponding improvement was 30.0%, 24.0%, 29.0%, 34.0%, and 4.4% on completion of one week therapy. On intergroup statistical comparison, mean improvement in pulmonary function test values were statistically significant or highly significant in regimens R1 to R4, as compared with placebo. However, improvement between any two regimens was not statistically significant in any of the regimens (R1-R4). Almost all the regimens were tolerated well and no patient showed major adverse reactions or cardiotoxicity necessitating withdrawal of the drug. On the other hand, minor adverse reactions were common and the high dose combination (R3) was found to have more adverse reactions than the low dose combination and either drug used alone. PMID- 8428856 TI - Bronchial hyperresponsiveness changes after 5 months of regular procaterol inhalation therapy in asthmatic children. AB - It is well known that procaterol has more potent antiallergic properties than previous beta-stimulants. Thirteen children with asthma who could be controlled for 2 months with a regular procaterol aerosol given 4 times/day (10 micrograms/dose) were studied. Histamine challenges were performed before and an average of 5 months after the consecutive treatment. No significant differences were seen in peak expiratory flow values. PC20 increased significantly from 132.9 +/- 127.7 micrograms/ml to 762.4 +/- 1205.7 micrograms/ml after 5 months. These results suggest that asthmatic children who can be controlled with regular use of the beta-stimulant procaterol are likely to have a decreased bronchial hyperresponsiveness. PMID- 8428857 TI - Increasing parental knowledge of asthma decreases the hospitalization of the child: a pilot study. AB - The parents of asthmatic children treated in the pediatric respiratory service of Wolfson Hospital were randomly divided into two groups. One group compromised 26 parents who voluntarily attended a teaching session. After completion of the course, a questionnaire was completed by parents who had attended the course and parents who had not. The first group showed greater knowledge of all aspects of the disease and, after a year's follow up, it was noted that there was a statistically significant lower hospitalization rate of their asthmatic child. PMID- 8428858 TI - Asthma and emotion: a review. AB - This review of the empirical literature on the relationship between asthma and emotion presents an explanatory model of the connection between them. Asthmatics tend to report and display a high level of negative emotion, and asthma exacerbations have been linked temporally to periods of heightened emotionality. Causality may be bidirectional. Hypothesized mediators for the relationship between asthma and emotionality include vagal and alpha-sympathetic hyperreactivity, predominant obstruction in the larger airways, individual response stereotypy, direct effects of emotion-related facial muscle tension on the airways, the emotional effects of asthma medications, heightened respiratory drive, and hyperventilation. Predictions are presented for research on this model of asthma and emotion, and for the psychological treatment of asthma. PMID- 8428859 TI - Efficacy and side effects of salbutamol in acute asthma in children: comparison of oral route and two different nebulizer systems. AB - Thirty-five separate episodes of acute bronchial asthma were studied in 21 asthmatic children. The bronchodilator, cardiovascular, and tremorogenic responses following three modes of administration of salbutamol were compared: open continuous nebulization (ON), closed-port intermittent nebulization (CN), and oral route (OR), for a period of 8 hours. Eleven acute attacks were treated by ON, 11 by CN, and 13 by OR. Pulmonary function was evaluated by clinical assessment and by the spirometric indices FEV1 and FEF25-75. Tremor was objectively measured, as well as heart rate (HR), respiratory rate (RR), and blood pressure (BP). Fastest onset of bronchodilator action, maximal response, and longest duration were seen when the drug was administered by the CN. Onset of tremorogenic effect was registered at 5 minutes when salbutamol was used by CN and ON and at 30 minutes when used by OR. During the first 30 minutes, tremor was significantly greater when salbutamol was used by CN than by OR. There were minor cardiovascular effects, except regarding HR after CN, which was significantly greater than after OR, at 5 and 30 minutes. Our findings indicate the inhaled route as the most effective route for administration of salbutamol to treat acute bronchospasm in children. The use of CN is a good alternative to jet nebulizers, but the greater dose of drug effectively administered by this system can briefly cause more tremor and heart acceleration. PMID- 8428860 TI - As-needed medication use in asthma usage patterns and patient characteristics. AB - Daily inhaled bronchodilator medication usage was recorded using an electronic device and airway obstruction by daily peak flow measurement. The demographic, clinical, and psychological characteristics of the subjects were noted. Subjects were allocated to as-needed (prn) medication usage groups according to the mean number of inhaler activations on days with zero, moderate, and severe airway obstruction. Segregation into arbitrary and nonarbitrary use, and into overuse, appropriate use, and underuse resulted in six usage groups. Appropriate use was observed in only 10 of 39 subjects. The major psychological variable to differ among groups was the MMPI variable Pt, representing general anxiety. Arbitrary users had a significantly higher mean score than nonarbitrary users. The variable Specific Internal Awareness, representing a perceived difficulty in recognizing the premonitory symptoms of an asthma attack, also differed among the usage groups, with arbitrary users having the lowest scores. These findings raise the possibility that reliance on an objective measurement of airway obstruction rather than on subjective symptomatology might enhance compliance with prn medication in some patients. PMID- 8428861 TI - Antidepressant treatments in children and adolescents. I. Affective disorders. AB - Part I of this review critically evaluates antidepressants' benefits for children and adolescents with affective disorders. The effectiveness of antidepressants is characterized with specific references regarding diagnostic methodology, measurement characteristics, response definitions, recovery rates, length of treatment, and plasma level monitoring. Antidepressants' efficacy for depressed youths is quite circumscribed, and their superiority to placebo remains unproved. Their intriguing lack of efficacy in affective disorders is discussed based on methodological issues and from a theoretical perspective. Clinical and research implications are presented. PMID- 8428862 TI - Suicidal children grow up: rates and psychosocial risk factors for suicide attempts during follow-up. AB - OBJECTIVE: Rates and psychosocial risk factors for suicide attempts during a 6 to 8-year follow-up period were compared for 25 predominantly prepubertal inpatient suicide attempters, 28 inpatient suicidal ideators, 16 nonsuicidal inpatients, and 64 nonpatients. METHOD: Standard research instruments were used to interview subjects and parents. Cox proportional hazard regression analyses were used to identify risk factors for a suicide attempt in the follow-up. RESULTS: No deaths occurred during follow-up. Suicide attempters were six times and suicidal ideators were three times more likely than were nonpatients to attempt suicide during follow-up. Poor social adjustment and mood disorder close to a recurrent suicide attempt were the strongest risk factors. CONCLUSIONS: Risk assessment should focus on identifying symptoms of mood disorders and impaired social adjustment in children with histories of suicide attempts and psychiatric hospitalization. PMID- 8428863 TI - Determinants for hospitalization from an emergency mental health service. AB - There has been limited systematic study of determinants of acute psychiatric hospitalization of children and adolescents. This study reviewed the records of children and adolescents who received emergency mental health services in one New Jersey county during 6 months of 1990 (N = 226). Using a structured form to abstract data, information was obtained on demographics, precipitating problems, past mental health services, substance use, family problems, and disposition. While suicidal behavior was not a predictor of acute hospitalization, the interaction of assaultive and suicidal behavior was predictive. Other contributory factors identified in the multivariate analysis included: child's substance use, family member's substance use, and initial emergency screening site. Recognition of present utilization patterns will facilitate the development of intensive community-based options for those with acute mental health problems. PMID- 8428864 TI - Utility of the head computerized tomography scan in child and adolescent psychiatry. AB - Despite the widespread use of computerized tomography (CT) in child and adolescent psychiatry, studies have not looked at its overall usefulness in a clinical setting. In this report, the authors examined 122 inpatients who had CT scans of the head. Only 27 patients had an abnormal CT scan. None of the patients had a change made in the diagnosis or treatment as a result of the scan findings. The authors suggest that routine CT scans of the head in child and adolescent inpatients is of limited value. In view of the rising costs of medical investigations, and the risk of irradiation to the developing brain, they suggest the need for clear indications for this procedure in clinical child and adolescent psychiatry. PMID- 8428865 TI - Effectiveness of a multimodal day treatment program for children with disruptive behavior problems. AB - OBJECTIVE: Children with disruptive behavior disorders are often also dysfunctional in academics, social skills, and self-esteem. The efficacy of multimodal day treatment in addressing these deficits was evaluated. METHOD: Thirty children, assigned to day treatment or a waiting list, were compared on measures of behavior, self-perception, academics, peer relations, and family functioning. RESULTS: Multivariate analyses of covariance showed that the treatment group improved significantly more on measures of behavior and self perception. Six-month follow-up findings indicate that treated children had improved over time on all measures except academics. CONCLUSIONS: This study demonstrates that compared with a waiting list control group, day treatment produces greater gains, and that these gains are maintained at 6 months' follow up. It also shows the need for a treatment that combines multiple modalities in dealing with children with disruptive behavior disorders. PMID- 8428866 TI - Private psychiatric hospitalization of children: predictors of length of stay. AB - OBJECTIVE: To quantitatively identify predictors and determinants of length of psychiatric hospital stay for children. METHOD: Forty-seven demographic, psychosocial stressor, psychopathology, and disposition variables were statistically reviewed as correlates of length of stay in 100 consecutive discharges from a child psychiatry inpatient service (age range 4-12) in a private hospital. Those with strong statistical significance were then analyzed by multiple regression. RESULTS: Greater severity of psychopathology (measured by the Children's Global Assessment Scale), greater severity of psychosocial stressors (by Axis IV scale), diagnosis of post-traumatic stress disorder, special educational and out-of-home dispositions, and severe tantrums in hospital all strongly predicted longer hospital stay. Diagnosis of adjustment disorder predicted shorter stay. Together these variables explained 57% of the total variance in length of stay. CONCLUSIONS: The most powerful of these predictor variables could potentially be measured at the time of admission, thus permitting accurate prediction of length of stay. A set of models was generated for this purpose. PMID- 8428867 TI - Use of inpatient services by a national population: do benefits make a difference? AB - This study describes changes in the use of inpatient mental health services by children and adolescents under age 18. The data were insurance claims from the Blue Cross and Blue Shield Federal Employees Program. The study focused on a cut in inpatient benefits that occurred between 1978 and 1983. The rate of inpatient hospitalization dropped significantly, and the average number of days also decreased significantly from 45.8 to 27.0 days. This study is among the first to demonstrate that the rate and amount of inpatient care provided for children and adolescents is responsive to variations in benefit coverage. PMID- 8428868 TI - The nosology of attention-deficit hyperactivity disorder. AB - This study examined five central issues regarding the classification of attention deficit hyperactivity disorder (ADHD) in an epidemiologically derived sample of primary school children. A monothetic schema (DSM-III) differed from a polythetic only schema (DSM-III-R) by having a higher frequency of comorbid conduct disorder. A pervasive model was too restrictive, identifying only the most severe disruptive or conduct disordered children. Raising the minimum threshold above eight symptoms (DSM-III-R) missed less disruptive children who nevertheless exhibit significant functional impairment. There was minimal support for the independence of a syndrome of attention deficit disorder without hyperactivity (DSM-III). Finally, ADHD patients commonly have other diagnoses, most frequently conduct disorder, oppositional defiant disorder, and anxiety disorder. PMID- 8428870 TI - The effects of a multimodal intervention with attention-deficit hyperactivity disorder children: a 9-month follow-up. AB - Using a double-blind, placebo design, we evaluated 96 attention-deficit hyperactivity disordered children for the effects of methylphenidate alone and in combination with behavioral parent training plus child self-control instruction. Seventy one of the children completed the treatment protocol. As reported previously, main effects were found for medication at posttest; however, there was no evidence of additive effects. Nine months after the termination of the behavioral interventions and the withdrawal of the stimulant medication, we found limited support for the hypothesis that the combined conditions would produce greater maintenance of treatment gains than would medication alone. PMID- 8428869 TI - Assessing parents' willingness to pursue treatment for children with attention deficit hyperactivity disorder. AB - OBJECTIVE: This study investigated parents' willingness to pursue treatment for attention-deficit-hyperactivity-disorder (ADHD). METHOD: A self-administered questionnaire (the ADHD Knowledge and Opinion Scale--AKOS) was developed and administered to a sample of 116 families attending an outpatient ADHD clinic. Socioeconomic status, parenting stress, family adaptability and cohesion, degree of child's externalizing behavior, and treatment history were obtained for each family. RESULTS: Parents' willingness to use medication or to pursue counseling were not related to family factors. History of the child receiving medication was mildly correlated with willingness to use medication. History of counseling was mildly correlated with willingness to use medication and to pursue counseling. Mothers who viewed their family as "enmeshed" reported significantly higher sense of competence than those viewing the family as "connected" or "disengaged." CONCLUSION: The AKOS is an instrument that may help clinicians identify and address parents' concerns about treatment for ADHD as well as parents' perceptions about their parenting skills. PMID- 8428871 TI - Does methylphenidate normalize the classroom performance of children with attention deficit disorder? AB - The present study examined the degree to which methylphenidate (MPH) normalized the classroom behavior and academic functioning of 31 children with attention deficit disorder (ADD). Subjects with ADD participated in a double-blind, placebo controlled trial in which children received each of four doses (5, 10, 15, and 20 mg) of MPH and a placebo following baseline measures. Children with ADD were compared with a group of 25 normal control children on teacher ratings of social conduct, direct observations of on-task behavior, and academic efficiency. At a group level of analysis, MPH exerted a significant effect on classroom measures of attention and academic efficiency to a point where they were no longer statistically deviant from scores obtained by normal control children. Nevertheless, when examined at the individual level, 25% of the sample failed to show normalized levels of classroom performance, thus implying the need for ancillary school-based interventions. PMID- 8428872 TI - A double-blind placebo controlled study of desipramine in the treatment of ADD: III. Lack of impact of comorbidity and family history factors on clinical response. AB - A 6-week randomized, double-blind, placebo controlled trial of desipramine (DMI) in daily doses averaging 4 to 5 mg/kg for the treatment of children and adolescents with attention deficit disorder with hyperactivity (ADDH) was further analyzed. Investigators examined whether comorbidity of ADDH with conduct disorder, major depression, an anxiety disorder, or a family history of ADDH predicted response to DMI treatment. There was a highly significant effect of treatment with DMI in outcome assessments, but responses to DMI were indistinguishable in ADDH patients with and without a comorbid disorder or familial ADDH. Cases of "pure" ADDH (lacking comorbidity with depression, anxiety, or conduct disorder and having a positive family history of ADDH) showed a trend toward lesser placebo responses and a corresponding greater DMI-placebo difference. These findings suggest that (1) DMI is effective in simple, noncomorbid cases, (2) DMI is not selective for comorbid cases, but (3) a response to DMI can be obtained even in complex cases of ADDH with associated comorbidity. PMID- 8428873 TI - Nortriptyline treatment of children with attention-deficit hyperactivity disorder and tic disorder or Tourette's syndrome. AB - Although as many as 50% of patients with Tourette's syndrome (TS) also meet diagnostic criteria for Attention-deficit Hyperactivity Disorder (ADHD), until recently little attention has been paid to ADHD symptoms in the assessment of therapeutic outcome of TS or patients with chronic motor tics (CMT). Because antipsychotics are of limited value in controlling the symptoms of ADHD and stimulants can exacerbate tics, alternative treatments for patients with chronic tic disorder (CTD) (TS or CMT) plus ADHD (CTD+ADHD) patients are direly needed. We examined the efficacy of the tricyclic antidepressant nortriptyline in the treatment of pediatric patients with CTD+ADHD ascertained from systematic chart reviews of all subjects with this diagnosis treated with nortriptyline. Of the 12 identified patients, 67% had significant improvement in CTD symptomatology and 92% significantly improved ADHD symptoms without major adverse effects over an average follow-up period of 19 months. Although the conclusions from this retrospective report can be only seen as preliminary until replicated in a controlled investigation, the magnitude and persistence of the response is encouraging and suggest a therapeutic role for nortriptyline in the treatment of CTD+ADHD patients. PMID- 8428874 TI - Childhood depression and risk of suicide: a preliminary report of a longitudinal study. AB - In the course of locating a sample of 427 adults who were assessed as children or adolescents with either major depressive disorder, mixed anxiety states, or no psychiatric disorder (normal controls), we found seven cases of suicide. Of the original sample, we located 159 of the 204 subjects with major depressive disorder (78%), 37 of the 66 subjects with anxiety disorders (56%), and 85 of the 177 normal controls (48%). All seven suicides occurred exclusively among the 159 children located from the major depressive disorder group, yielding a rate of 4.4% over approximately 10 years. Psychological autopsy was conducted in the seven suicide victims to assess the psychological status since the initial assessment and at the time of death. Although the onset of the first depressive episode in these victims was around puberty, the suicides usually did not occur until late adolescence or early adulthood. At least five of the seven subjects had recurrent depressive symptoms and were clinically depressed at the time of death. These preliminary findings suggest that major depressive disorder in childhood has significant mortality by suicide. PMID- 8428875 TI - Bupropion exacerbates tics in children with attention-deficit hyperactivity disorder and Tourette's syndrome. AB - Concerns have been raised regarding the development of tics in some children with attention-deficit hyperactivity disorder (ADHD) receiving stimulants. Because in many patients with Tourette's syndrome (TS) the ADHD symptoms are the major source of disability, alternative nonstimulant treatments are needed. Initial reports on bupropion have suggested that it may be an effective alternative in children and adults with ADHD. The value of bupropion as an alternative treatment for patients with comorbid TS and ADHD is determined by the question of association with exacerbation of tics. The purpose of this study was to examine an apparent association with exacerbation of tics in patients with comorbid TS and ADHD. A careful retrospective analysis was conducted of clinic cases of patients who all had comorbid TS and ADHD treated with bupropion. We present four cases of children with ADHD and comorbid TS treated with bupropion in whom tics were exacerbated by this medicine. This series suggests that bupropion may not be an appropriate alternative to stimulants in the treatment of ADHD in TS. PMID- 8428876 TI - Resolved: two-week psychiatric hospitalizations of children and adolescents are useless. PMID- 8428877 TI - From abuse to violence. PMID- 8428878 TI - Clozapine for schizophrenia. PMID- 8428879 TI - New TS treatment. PMID- 8428880 TI - Parental expectations and test anxiety. PMID- 8428881 TI - The outcome of adolescent depression in the Ontario Child Health Study follow-up. AB - Outcome data from a longitudinal community-based study are presented for 652 adolescents who had a major depressive syndrome (MDS), conduct disorder (CD), both disorders, or neither disorder at the study inception. In general, it was found that both MDS and CD carried with them significant psychiatric and psychosocial morbidity 4 years later. Statistical comparisons between the MDS and CD groups were not significant for any outcome but were suggestive of distinct outcomes for these two disorders. Findings for the "mixed" MDS and CD group are presented and the nosological status of this group is discussed. PMID- 8428882 TI - The course of major depressive disorder in adolescents: I. Recovery and risk of manic switching in a follow-up of psychotic and nonpsychotic subtypes. AB - Fifty-eight adolescents meeting Research Diagnostic Criteria (RDC) for major depressive disorder were observed for 24-months after inpatient admission in a prospective, naturalistic design study to assess time to recovery, risk of manic switching, and level and persistence of psychosocial impairment. Subjects were further categorized as psychotic or nonpsychotic by RDC. The cumulative probability of recovery by 24 months was high (90%); however, the episode was protracted with only 6.9% recovering within 12 weeks and only 29.3% recovering by week 20. Time to recovery did not vary by RDC subtype, but manic switching was observed only among psychotics, at a rate of 28%. Psychotics were more likely than were nonpsychotics to exhibit psychosocial impairment through 6 months of follow-up, but this difference narrows by 24 months. Implications of these findings for understanding developmental effects on the course and treatment of depression in adolescents are discussed. PMID- 8428883 TI - Effect of tricyclic antidepressants on switching to mania and on the onset of bipolarity in depressed 6- to 12-year-olds. AB - The authors present data on the rates of onset of bipolar phenomena, at 2- to 3 year follow-up, in depressed 6- to 12-year olds. The subjects had participated in the nortriptyline drug study. There were high rates of onset of bipolarity and of switching to mania while patients were on tricyclic antidepressants (TCAs). Mania developed only in subjects who had received TCAs at some time in the past or were receiving them concurrent with the onset of mania. These findings were analyzed with respect to the influence of multiple covariates, including family history of manic disorders and pubertal status. The authors discuss the implications of these findings for the prescription of TCAs to children who present with major depressive disorder and have a family history of bipolarity or a history of bipolar symptoms. The relevance of these findings for the later development of rapid cycling is discussed and compared with predictors of rapid cycling in adults. PMID- 8428884 TI - The specificity of psychological characteristics of adolescent suicide attempters. AB - Using a semistructured interview, psychological characteristics were gathered from three groups of adolescents: 48 suicide attempters, 66 depressed, and 43 nondepressed adolescents who never attempted suicide. Various characteristics reported in literature discriminated the attempters from the normal group but not from the depressed group. These characteristics probably are not specific for adolescent suicide attempters. Even when a discriminant analysis was applied, the suicide attempters showed many similarities with the depressed adolescents, although both these groups could be separated from the normal group. Using follow up data, the quadrants created by the axes appeared to have more implications for prevention than for the a priori group membership. PMID- 8428885 TI - Psychosocial characteristics of adolescents with a history of suicide attempt. AB - The purpose of this study was to identify psychosocial risk factors uniquely associated with past suicide attempts. Data assessing a large number of variables were available from a representative sample of older adolescents (N = 1,710). Most independent variables were associated with past suicide attempts; variables that remained associated with past attempts after controlling for current depression level included externalizing and internalizing problem behaviors, past psychiatric disorders, depressotypic cognitions, coping, school problems, health problems, and gender. The probability of having made an attempt increased dramatically as a function of the number of risk factors. Females had more risk factors and showed a greater vulnerability to the factors than did males. PMID- 8428886 TI - Personality disorder, tendency to impulsive violence, and suicidal behavior in adolescents. AB - Inpatient suicide attempters (n = 37) were compared with never-suicidal psychiatric controls (n = 29) with respect to prevalence and severity of personality disorder, history of aggression and assaultive behavior, and other measures of impulsive violence. Attempters, compared with controls, were more likely to show evidence of personality disorder or trait, particularly those of the borderline type. Attempters showed greater number of borderline symptoms than did controls, even after removing the item relating to suicidality. There were no differences between the groups with respect to lifetime history of aggression, history of assaultive behavior, or self-report measures of a tendency to impulsive aggression. Those attempters with personality disorder were much more likely to have made a previous attempt. The apparently high prevalence of personality disorder among adolescent inpatient suicide attempters indicates that the social impairment associated with personality disorder be viewed as an important aspect of treatment. PMID- 8428887 TI - Friends of adolescent suicide attempters and completers. AB - Friends of adolescent suicide attempters and suicide completers were compared with students who had low exposure to suicide on a range of preexposure, concurrent, and outcome variables. Subjects who were friends of both suicide attempters and completers (N = 84) differed significantly from the low-exposure group (N = 554) on most variables examined. Friends of suicide completers only (N = 68) and attempters only (N = 92) did not differ from one another; however, the latter group reported significantly higher levels of current depression and suicidal behavior than the low-exposure group. Greater attention needs to be paid to friends of suicide attempters in the development and evaluation of suicide prevention strategies. PMID- 8428888 TI - Suicidal behaviors and childhood-onset depressive disorders: a longitudinal investigation. AB - In this longitudinal study, the rates and correlates of suicidal ideation and suicide attempts were determined among outpatient youths with depressive disorders and youths with other psychiatric disorders. At study entry, about 66% of the subjects evidenced suicidal ideation and 9% already attempted suicide. The rate of ideation remained fairly stable over time, whereas the rate of attempts reached 24% by the average age of 17 years. Major depressive and dysthymic disorders were associated with significantly higher rates of suicidal behaviors than were adjustment disorder with depressed mood and nondepressive disorders. In the presence of affective disorders, comorbid conduct and/or substance use disorders further increased the risk of suicide attempts. PMID- 8428889 TI - Alcohol consumption in relation to other predictors of suicidality among adolescent inpatient girls. AB - This study of 54 adolescent inpatient girls examined alcohol consumption in relation to depression severity and family dysfunction as predictors of suicidal ideation and behavior. Although alcohol consumption, depression severity, and family dysfunction were intercorrelated, regression analyses revealed their differential importance to the prediction of self-reported suicidal ideation and severity of clinician-documented suicidal ideation or behavior (none, ideation, intent, gesture, attempt). Self-reported ideation was strongly predicted by depression severity and family dysfunction; severity of clinician-documented suicidal ideation or behavior was predicted by alcohol consumption and family dysfunction. Implications for assessment and treatment are discussed. PMID- 8428890 TI - Are adolescent suicide attempters noncompliant with outpatient care? AB - The outpatient clinic attendance patterns of 115 consecutively referred 10- to 18 year-old suicide attempters and of 110 nonattempters were compared. The two groups did not differ in number of appointments scheduled or missed, but attempters kept significantly fewer appointments than did nonattempters. Seventy seven percent of each group dropped out of treatment, but attempters dropped out significantly faster. Attendance and dropout were unrelated to age, reason for referral, or previous attempts. Girls missed more appointments than did boys, and Hispanic patients kept a smaller percentage of scheduled appointments than did other ethnic groups. We conclude that adolescent attempters are not more likely to drop out of treatment but keep fewer appointments and remain in care more briefly than do other outpatients. Recommendations for triage and brief case management are made. PMID- 8428891 TI - Adolescent psychiatric inpatients' risk of suicide attempt at 6-month follow-up. AB - Adolescent inpatients (of whom 48 were admitted for a suicide attempt, 33 were admitted for suicidal ideation, and 53 had no history of clinically significant suicidal ideation or attempt), were interviewed while in the hospital and then followed up 6 months later. Of the 134 patients followed up, 13 (9.7%) had made a suicide attempt. The vast majority of those who attempted suicide had been suicidal while in the hospital (12/13 or 92.3%). Other risk factors for suicidal behavior include major depression at intake, affective disorder with nonaffective comorbidity, a depressive disorder that continued through follow-up, death of a relative, and family financial problems. Suicidal inpatients, particularly those with chronic and recurrent affective illness, are at substantial risk for making a suicide attempt within 6 months of discharge. At follow-up, an even higher proportion showed attempts or suicidal ideation with a plan (N = 36 or 26.8%), with risk factors similar to those noted above. More intense outpatient or partial hospital interventions as a transition from the inpatient environment may be necessary to reduce the rate of recidivism among suicidal adolescents. PMID- 8428892 TI - Psychotropic treatment of chronic fatigue syndrome and related disorders. AB - BACKGROUND: Chronic fatigue syndrome (CFS) and fibromyalgia frequently are associated with symptoms of major depression. For this reason, antidepressants have been used in treatment of these disorders; however, little direction has been provided into this application in psychopharmacology. METHOD: First, nine studies were reviewed regarding the relationship of the symptoms of fatigue and depression. Next, 23 reports (12 double-blind studies, 7 open studies, and 4 case reports) were reviewed for the effectiveness of therapy as assessed by global response and improvement of both depression and pain. Studies were differentiated by type of controls, as well as by alleged mechanism of action of the pharmacologic agent. RESULTS: Disturbances in brain neurochemistry shared by CFS and major depression may serve as a basis for the effectiveness of some antidepressants in CFS. Response to some antidepressants in patients with CFS or fibromyalgia may occur at doses lower than those used in major depression, e.g., amitriptyline 25-75 mg/day. We further found that the more serotonergic treatments (e.g., clomipramine) were more successful in alleviating pain than depression, whereas catecholaminergic agents (e.g., maprotiline, bupropion) seemed particularly effective for symptoms of associated depression. CONCLUSION: To maximize response of the physiologic and psychological consequences of the disorder, more investigation is needed to replicate the apparent findings that relate the neurochemical impairment underlying CFS and fibromyalgia to the type of antidepressant mechanism. PMID- 8428893 TI - Conduct disorder and personality disorders in hospitalized adolescents. AB - BACKGROUND: The purpose of this study was to investigate the differences between DSM-III-R Axis I and II psychiatric diagnoses in hospitalized adolescents with and without conduct disorder. The spectrum of psychopathology associated with conduct disorder, especially personality disorder symptoms and diagnoses, remains largely unexplored. METHOD: Twenty-five inpatients were evaluated using the Diagnostic Interview for Children and Adolescents, Adolescent Version (DICA-R-A); the Schedule for Affective Disorders and Schizophrenia for School-Age Children, Epidemiologic Version (K-SADS-E) (panic disorder and agoraphobia only); and the Structured Interview for DSM-III-R Personality Disorders (SIDP-R). RESULTS: Fifty two percent met criteria for conduct disorder, and the majority of these had comorbid substance abuse, attention-deficit hyperactivity disorder, and major depression. The diagnosis of conduct disorder was also associated with fulfilling criteria for multiple Axis II personality disorders, particularly passive aggressive and histrionic. Nearly three personality disorders per subject were found in those with conduct disorder, while those without conduct disorder averaged just over one personality disorder. There was a trend for female subjects with conduct disorders to have more frequent and varied personality disorder diagnoses. For the entire sample, males were significantly more likely to have conduct disorder, and females were significantly more likely to have borderline personality disorder. CONCLUSION: These findings support conduct disorder as a multifaceted illness composed of diverse Axis I and II characteristics. Long-term follow-up studies are needed to determine the outcome of personality disorder diagnoses made in adolescents with and without conduct disorder. PMID- 8428894 TI - Fluoxetine efficacy in social phobia. AB - BACKGROUND: To determine the efficacy of fluoxetine in the treatment of social phobia, fluoxetine was administered to 16 patients with a primary DSM-III-R diagnosis of social phobia in a 12-week, open, clinical trial. METHOD: Treatment began at 20 mg of fluoxetine daily and was increased according to clinical response and side effects every 4 weeks. Patients completed self-report measures of anxiety and depression at baseline and at Weeks 4, 8, and 12. These included the Beck Depression Inventory, the Social Avoidance and Distress Scale, the Fear of Negative Evaluation Scale, the State-Trait Anxiety Inventory, the Fear Questionnaire, the Social Anxiety Thoughts Questionnaire, and the Social Adjustment Scale-Self-Report. Clinicians completed a Clinical Global Improvement Scale. RESULTS: Thirteen of the 16 patients completed the trial. Three patients (18.8%) were unable to complete the trial due to adverse side effects. Of the 13 (81.2%) remaining patients, 10 were considered to be responders and 3 were considered to be nonresponders. Measures of social anxiety and phobic avoidance showed a significant improvement from baseline to endpoint, achieving a significance of p < .005. Responders to fluoxetine were more likely to have been older at onset of the social phobic symptoms and had a shorter duration of illness. CONCLUSION: These findings suggest that fluoxetine may be effective in the treatment of social phobia. Double-blind studies will be required to further investigate these findings. PMID- 8428895 TI - Feasibility of an every-other-night regimen in insomniac patients: subjective hypnotic effectiveness of quazepam, triazolam, and placebo. AB - BACKGROUND: Rebound insomnia, a worsening of sleep difficulty beyond baseline levels, can complicate the physician's attempt to use regularly scheduled drug holidays in the management of insomniac patients. Quazepam, a benzodiazepine with a long half-life, has been shown to exhibit carryover effectiveness for the first night or two following withdrawal. This finding suggests a potential utility for an alternate-night drug regimen in which the withdrawal features of the compound serve as a potential benefit. METHOD: A randomized, double-blind, three compartment, parallel-group design of 5 weeks' duration, comparing quazepam 15 mg, triazolam 0.5 mg, and placebo, was conducted in 65 insomniac subjects. This study was a nonpolysomnographic study utilizing sleep questionnaires. RESULTS: No differences were noted between quazepam and triazolam on treatment nights. Evidence of carryover effectiveness with quazepam and rebound effects with triazolam were noted on off-treatment nights. CONCLUSION: The efficiency of alternate-night therapy with quazepam should be rigorously evaluated using polysomnographic determinations. PMID- 8428896 TI - Cyclobenzaprine-induced delirium in two octogenarians. PMID- 8428897 TI - Adventitious change in homosexual behavior during treatment of social phobia with phenelzine. PMID- 8428898 TI - Atypical antidepressants versus imipramine in the treatment of major depression: a meta-analysis. AB - BACKGROUND: Our investigation involved a quantitative literature review technique known as meta-analysis to compare the efficacy of three newer antidepressants and imipramine. METHOD: We examined seven major journals in psychiatry from 1980 through 1990, inclusive, and selected those investigations of imipramine, trazodone, bupropion, and fluoxetine that met our minimal criteria for interpretability. These criteria included: (1) the presence of a placebo control, (2) double-blind status, (3) the use of the Hamilton Rating Scale for Depression as a dependent variable measure, (4) the use of nongeriatric adults with a diagnosis of major depression by DSM or RDC standards, and (5) the presence of reported means and standard deviations in the investigation, or sufficient data that allowed such to be computed. Each study of four antidepressants was analyzed for an effect size of the drug investigated. The effect size allows for a determination of the efficacy of a particular drug as compared with placebo, measured in standard deviation units. RESULTS: The data indicated that all four agents are effective as compared with placebo. Furthermore, there is no evidence that the newer heterocyclic agents are less effective than imipramine, as an ANOVA showed no statistically significant difference between the effect sizes of the four antidepressants. CONCLUSION: These data are discussed in terms of characteristics of the various investigations and the need for further research comparing the efficacy of psychopharmacologic agents. PMID- 8428899 TI - In vivo generation of an adenylylcyclase isoform with a half-molecule motif. AB - A truncated form of adenylylcyclase (type V-alpha) has been cloned from a cardiac cDNA library. It constitutes a half-molecule of type V adenylylcyclase diverging at the end of the first cytoplasmic loop. Northern blotting study has revealed the presence of such a mRNA species (approximately 3.5 kilobases in size) in the heart. Genomic sequence analysis has revealed that type V-alpha is generated via usage of a polyadenylation signal located within an intronic sequence of type V adenylylcyclase gene. When type V-alpha is co-expressed with an artificially generated half-molecule constituting the latter half of type V adenylylcyclase, the catalytic activity in transfected cell membranes is significantly higher than that of controls. However, when either alone is overexpressed, no significant increase in catalytic activity results. These results indicate that a half molecule of adenylylcyclase, i.e. a protein containing six-transmembrane spans followed by a single cytoplasmic domain, can be generated in vivo, but catalytic activity is lacking unless heterodimerization can occur. This finding identifies another potential mechanism for generating diversity within this enzyme family. PMID- 8428900 TI - Protein-tyrosine kinase p72syk is activated by thrombin and is negatively regulated through Ca2+ mobilization in platelets. AB - Activation of platelets by thrombin results in a dramatic increase in tyrosine phosphorylation on multiple cellular proteins (Ferrell, J. E., and Martin, G. S. (1988) Mol. Cell. Biol. 8, 3603-3610; Golden, A., and Brugge, J. S. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 901-905; Nakamura, S., and Yamamura, H. (1989) J. Biol. Chem. 264, 7089-7091). However, none of the responsible protein-tyrosine kinase has been reported so far. We report here that p72syk, one of the non receptor-type protein-tyrosine kinases, is activated following thrombin stimulation in blood platelets. Washed porcine platelets were stimulated by thrombin, and the activation of p72syk was assessed in an immunoprecipitation kinase assay. The activity of p72syk increased within 5 s, reached a maximum at 10 s, and decreased to a basal level within 60 s after 0.5 unit/ml thrombin stimulation. The amount of immunoprecipitated p72syk was not altered throughout the time course. This activation was greatly enhanced in a dose-dependent manner and was completely canceled by the pretreatment of platelet suspension with hirudin, a specific antagonist of thrombin. In the Ca(2+)-depleted condition both extra- and intracellularly, the activation of p72syk was still persistent; in contrast, the deactivation process was completely abrogated even at 120 s after thrombin stimulation. In addition, the replenishment of Ca2+ resulted in a similar deactivation pattern as seen in the Ca(2+)-rich condition. Furthermore, this deactivation was also canceled by the pretreatment of platelets with W7, a calmodulin antagonist, as well as ML9, a myosin-light-chain kinase inhibitor. These results indicate that p72syk can be a responsible enzyme to the protein tyrosine phosphorylation events following the platelet activation by thrombin and may be negatively regulated by Ca2+ in a calmodulin-dependent manner, inter alia myosin light-chain kinase, in thrombin-stimulated platelets. PMID- 8428901 TI - Direct interaction between the transcriptional activation domain of human p53 and the TATA box-binding protein. AB - The human p53 tumor suppressor gene product can activate transcription by RNA polymerase II in the yeast, Saccharomyces cerevisiae, as well as in human cells. Several viral transcriptional activator proteins have been shown to directly contact TBP, the TATA box-binding subunit of the general initiation factor, TFIID. In this report, we use protein affinity chromatography to show that the cellular transcription factor, p53, interacts directly and specifically with yeast TBP. The TBP binding domain of p53 was localized to its N-terminal 73 amino acids. This highly acidic portion of p53 functions as a transcriptional activation domain and is deleted in some tumors induced by the Friend leukemia virus. A human tumor-derived oncogenic point mutation of p53, which lies outside the activation domain of p53, but reduces its ability to activate transcription, greatly reduced the ability of p53 to bind yeast TBP in vitro. This mutation probably affects the overall conformation of the protein and indirectly interferes with the ability of p53 to contact TBP and activate transcription. In contrast, a mutated oncogenic form of p53 that is unaffected in its ability to activate transcription bound yeast TBP as well as wild type p53. The human TBP activity in a HeLa extract also bound to the activation domain of p53. Our data support a general model in which DNA-bound activator proteins activate transcription by interacting with TBP. PMID- 8428902 TI - T4-phage deoxycytidylate deaminase is a metalloprotein containing two zinc atoms per subunit. AB - Deoxycytidylate (dCMP) deaminase, a hexameric allosteric enzyme induced on infection of Escherichia coli by bacteriophage T4, was shown to contain two atoms of zinc per subunit by atomic absorption spectroscopy. One zinc appears to be involved in catalysis, as described for adenosine deaminase (Sharaff, A. J., Wilson, D. K., Chang, Z., and Quiocho, F. A. (1992) J. Mol. Biol. 226, 917-921) and cytidine deaminase (Yang, C., Carlow, D., Wolfenden, R., and Short, S. A. (1992) Biochemistry 31, 4168-4174). This thesis is supported by the finding that the enzyme loses about 80% of its activity in the presence of o-phenanthroline. It has also been found that zinc is released when the enzyme is denatured in the presence of the metallochromic indicator, 4-(2-pyridylazo)resorcinol. Renaturation of the deaminase to an active form occurred in the presence but not in the absence of zinc. The second atom of zinc is proposed to be located in a region of T4-dCMP deaminase that resembles a zinc finger. This region, which has the sequence His-X3-Cys-X14-His-X3-His, would represent a zinc-binding motif that has not been described previously. PMID- 8428903 TI - Formyl peptide receptor chimeras define domains involved in ligand binding. AB - We have begun to study the structural requirements for the binding of formyl peptides to their specific receptors. As an initial approach, we constructed C5a formyl peptide receptor chimeras. Unique (and identical) restriction sites were introduced within the transmembrane domains of these receptors that allowed for the exchange of specific areas. Four types of chimeric receptors were generated. 1) The C5a receptor was progressively substituted by the formyl peptide receptor. 2) The formyl peptide receptor was progressively substituted by the C5a receptor. 3) Specific domains of the C5a receptor were substituted by the corresponding domain of the formyl peptide receptor. 4) Specific domains of the formyl peptide receptor were replaced by the same corresponding domain of the C5a receptor. Wild type and chimeric receptors were transfected into COS 7 cells and their ability to bind formyl peptide determined, taking into account efficiency of transfection and expression of chimeric protein. Based on these results, a ligand binding model is presented in which the second, third, and fourth extracellular (and/or their transmembrane) domains together with the first transmembrane domain form a ligand binding pocket for formyl peptides. It is proposed that the amino-terminal domain plays a role by presumably providing a "lid" to the pocket. The carboxyl terminal cytoplasmic tail appears to modulate ligand binding by regulating receptor affinity. PMID- 8428904 TI - Adenovirus-mediated augmentation of cell transfection with unmodified plasmid vectors. AB - The present study demonstrates that the human adenovirus (Ad) can augment transfer and expression of a gene within plasmid DNA unmodified by nonspecific linkers or by linker-ligand complexes. Following the transfection of COS-7 cells with pRSVL, a luciferase expression plasmid vector directed by the Rous sarcoma virus-long terminal repeat promoter, luciferase activity in the target cells was 10(3)- to 10(4)-fold higher when the cells were also infected with Ad-CFTR, a replication-deficient recombinant Ad containing human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA. The enhancement of luciferase gene expression in COS-7 cells was also observed with Ad-dl312 (a replication deficient E1a deletion mutant Ad with no exogenous gene) and wild type Ad5. The efficiency of cell transfection with pRSVL in the presence of an Ad was achieved in a dose-dependent fashion with progressively higher luciferase activity in cells infected by increasing amounts of Ad-CFTR, Ad-dl312, or Ad5. The augmentation by Ad-CFTR of the transfer and expression of the luciferase gene in cells was similar to that of another transfection reagent, cationic liposomes. Further, when Ad-CFTR and liposomes were used in combination, 4- to 100-fold more efficient expression of the luciferase gene was achieved than with Ad-CFTR or liposomes alone. When COS-7, HeLa, and CV-1 cells were evaluated in parallel in the presence or absence of liposomes, Ad-mediated enhancement of luciferase activity was observed in all cell lines. Thus, exposure of target cells to replication-deficient or competent human Ad will markedly augment transfer and expression of the genes within plasmid DNA in mammalian cells in vitro without modifying the plasmid with linkers or linker-ligand complexes, a strategy that should be useful for in vitro and in vivo gene transfer applications. PMID- 8428905 TI - Guanosine tetraphosphate inhibits protein synthesis in vivo. A possible protective mechanism for starvation stress in Escherichia coli. AB - Guanosine 3',5'-bispyrophosphate (ppGpp) accumulates in bacteria in response to either amino acid or energy source starvation. Here we demonstrate that levels of ppGpp similar to those induced by amino acid starvation inhibit the rate of protein synthesis by 84-91%. The intracellular concentration of ppGpp is manipulated in our studies by increasing the expression of a truncated relA gene encoding a smaller but catalytically active peptide with ppGpp synthetase activity. We find that the intracellular activity of the truncated RelA peptide is insensitive to chloramphenicol, unlike the product of the wild-type relA gene, ppGpp synthetase I. Previously, this same ppGpp expression system was used (Schreiber, G., Metzger, S., Aizenman, E., Roza, S., Cashel, M., and Glaser, G. (1991) J. Biol. Chem. 226, 3760-3767) to demonstrate that increasing the ppGpp concentration inhibits growth and ribosomal RNA transcription, and they found suggestive evidence for ppGpp inhibition of protein synthesis. We further investigated the effect of ppGpp on protein synthesis and find that ppGpp is a potent inhibitor of protein synthesis as well as glycerol accumulation but has no effect on transport of methionine, the amino acid used in measuring protein synthesis rates, or on uptake of alpha-methylglucoside, a non-metabolizable analogue of glucose. PMID- 8428906 TI - Expression of rat liver reduced glutathione transport in Xenopus laevis oocytes. AB - We have studied the expression of the hepatic GSH transport system in Xenopus laevis oocytes. Injection of rat liver poly(A)+ RNA resulted in the functional expression of the GSH transport system determined as GSH efflux from GSH loaded oocytes. Expression required 3-5 days to process the liver mRNA. Methionine, cystathionine, and sulfobromophthalein (BSP)-GSH inhibited the efflux of GSH from liver mRNA-injected oocytes according to their known cis or transactions on hepatocytes, namely BSP-GSH from inside and methionine and cystathionine from outside. The expressed hepatic GSH transport system also mediated the uptake of intact GSH into the oocyte, consistent with the bidirectional operation of this facilitative transporter. The uptake of GSH into mRNA-injected oocytes was inhibited by BSP-GSH in chloride-free conditions. Finally, two different mRNA size fractions encoded for hepatic GSH transport activity (uptake or efflux): a 2.0-2.5-kilobase size class, which expressed GSH transport (uptake or efflux) completely inhibited by BSP-GSH (compatible with sinusoidal GSH transport), and a 3.5-4.0-kilobase size class, which expressed GSH transport (uptake or efflux) not inhibited by BSP-GSH. These results demonstrate that hepatic GSH transport can be expressed in Xenopus oocytes and mRNA of two distinct sizes encode for GSH transporters. PMID- 8428907 TI - Molecular cloning and sequence of a novel ommochrome-binding protein cDNA from an insect, Manduca sexta. AB - An ommochrome-binding protein (OBP) from the hemolymph of Manduca sexta has recently been purified and characterized. A cDNA clone was isolated from a fifth instar larval cDNA expression library utilizing antiserum against OBP. Northern blot analysis of total fat body RNA detected a transcript of approximately 1.2 kilobases in fifth instar wandering larvae RNA. The complete nucleotide sequence of the 905-base pair cDNA insert was determined by the dideoxy chain termination method. The OBP cDNA encodes a polypeptide of 274 residues with a predicted molecular weight of 30,580 and with one consensus N-linked glycosylation site. Comparison of the NH2-terminal sequence of the mature protein and the cDNA sequence revealed a typical signal peptide of 18 amino acids. In wandering stage larvae, the OBP transcript appeared to be at least 250-fold less abundant than ribosomal RNA. PMID- 8428909 TI - N-myristylation of the catalytic subunit of cAMP-dependent protein kinase conveys structural stability. AB - Coexpression of the yeast N-myristyltransferase with the murine catalytic subunit of cAMP-dependent protein kinase in prokaryotic cells results in the N myristylation of the recombinant catalytic subunit. The acylated recombinant catalytic subunit was purified following in vitro holoenzyme formation with a mutant form of the regulatory subunit and compared to the non-myristylated recombinant enzyme and to the mammalian porcine enzyme. All three enzymes are very similar in terms of their kinetic properties and their capacity to reassociate in vitro with the regulatory subunit to form holoenzyme. In contrast, the myristylated recombinant catalytic subunit is significantly more stable to thermal denaturation than the non-myristylated enzyme. Its thermal stability is now comparable to the mammalian enzyme. All three catalytic subunits are significantly more stable to thermal denaturation when they are part of the holoenzyme complex. Each shows an increase in T1/2 of 10 degrees C. This study demonstrates that one function for the myristic acid at the NH2 terminus of the catalytic subunit is to provide structural stability. PMID- 8428908 TI - Differential effects of brefeldin A on transport of secretory and lysosomal proteins. AB - Brefeldin A (BFA) rapidly blocks anterograde exocytotic transport through the Golgi complex. Sustained retrograde traffic induced by brefeldin A causes redistribution of constituents of the Golgi, but not the trans-Golgi network (TGN), to the endoplasmic reticulum (ER). In the present study on HepG2 cells, we have observed a differential effect of BFA on transport from the TGN of two soluble proteins: alpha 1-antitrypsin as a representative of secretory proteins and cathepsin D as a prototype of lysosomal enzymes. The Golgi complex of HepG2 cells is sensitive to BFA, as within minutes after its addition nearly all activity of three resident Golgi enzymes was recovered in the ER as monitored by cell fractionation on sucrose density gradients. In accordance with this, "high mannose"-glycosylated alpha 1-antitrypsin was retained in or transported back to the ER. "Complex"-glycosylated alpha 1-antitrypsin was neither secreted into the medium nor transported back to the ER. Most of it was retained in vesicles with the buoyant density of Golgi. These vesicles contained the fluid phase endocytotic marker horseradish peroxidase when this was added to the culture medium prior to the BFA, suggesting that the vesicles derived from the TGN. After BFA addition, the compartment became inaccessible to endocytosed horseradish peroxidase. In contrast to blocking transport of complex alpha 1-antitrypsin, BFA did not affect processing of newly synthesized complex-glycosylated procathepsin D (53 kDa) to the mature 31-kDa form. Neither did it interfere with processing of endocytosed procathepsin D. That the mature cathepsin D had indeed reached the lysosomes was verified by Percoll density gradient fractionation. In conclusion, in HepG2 cells, BFA induces two blocks in the secretory pathway: one at the level of the ER-Golgi juncture and the other in the TGN. In contrast, transport from the Golgi complex to the lysosomes and from the plasma membrane to the lysosomes continued. PMID- 8428910 TI - Import and assembly of the beta-subunit of chloroplast coupling factor 1 (CF1) into isolated intact chloroplasts. AB - The transit peptide gene of the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from Nicotiana plumbaginifolia was fused to the tentoxin resistant beta-subunit gene of chloroplast coupling factor 1 (CF1) from Nicotiana tabacum via a linker sequence. The consequent fusion gene encodes the entire gene sequences of both the transit peptide and the beta-subunit of CF1 without a single change of amino acid residues. The fusion gene was in vitro expressed in a coupled transcription-translation system as a 62-kDa precursor and was imported into isolated intact chloroplasts of Nicotiana longiflora. The imported precursor was found to be processed to the expected mature beta-subunit size. Evidence is presented that radioactively labeled beta-subunit was incorporated into mature CF1 and not just nonspecifically associated with the thylakoid membranes. Since the fusion protein containing only the transit peptide sequence was imported and apparently correctly processed, it may not be necessary to include N-terminal amino acids of the mature small subunit for correct proteolytic cleavage in the chloroplast stroma. PMID- 8428911 TI - Gs mediates hormonal inhibition of the calcium pump in liver plasma membranes. AB - We have reported that the calcium pump in liver plasma membranes is coupled to Gs or a Gs-like protein. However, we show here that isoproterenol, which activated adenylyl cyclase via Gs, had no effect on the calcium pump, while human calcitonin, human parathyroid hormone, and mini-glucagon, which inhibited this system, did not affect adenylyl cyclase activity. In order to determine the nature of the G protein coupled to the calcium pump, we used the RM antibody, raised against the carboxyl-terminal decapeptide of Gs alpha, which antagonized adenylyl cyclase activation by isoproterenol or glucagon. The RM antibody specifically blocked calcium pump inhibition by mini-glucagon, calcitonin, or parathyroid hormone, while it did not affect guanosine 5'-O-(thiotriphosphate) inhibition. Its effect was mimicked by the corresponding decapeptide RMHLRQYELL. The AS/7 antibody, reactive with Gt alpha, Gi 1 alpha, and Gi2 alpha, was ineffective. Complementation of liver plasma membranes with in vitro translated Gs alpha-2, the large form of Gs alpha, led to a 40% decrease in calcium pump activity, with a parallel 2-fold increase in adenylyl cyclase activity. In vitro translated Gi1 alpha did not affect the calcium pump activity, while it evoked a 40% inhibition of adenylyl cyclase activity. We conclude that a same Gs alpha may be coupled either to the calcium pump or to adenylyl cyclase. However, Gs is functionally specialized, since it does not ensure cross-talk between the two receptor-effector systems. These results point out the possible compartmentalization of Gs. PMID- 8428912 TI - Actin-binding peptides obtained from the C-terminal 24-kDa fragment of porcine aorta smooth muscle myosin subfragment-1 heavy chain. AB - The C-terminal 24-kDa fragment of myosin subfragment-1 (S-1) heavy chain was isolated by papain digestion of porcine aorta myosin followed by ethanol fractionation. The isolated 24-kDa fragment was digested by lysylendopeptidase. When the digest was ultracentrifuged with F-actin, 7- and 2-kDa peptides coprecipitated with the actin. These two peptides were isolated by high performance liquid chromatography, and their amino acid sequences were determined. The 7- and 2-kDa peptides correspond to residues 692-744 and 835-846, respectively, of the chicken gizzard myosin heavy chain (Yanagisawa, M., Hamada, Y., Katsuragawa, Y., Imamura, M., Mikawa, T., and Masaki, T. (1987) J. Mol. Biol. 198, 143-157). The 7-kDa peptide contains the reactive cysteine residues, SH1 and SH2. The isolated 7- and 2-kDa peptides also bound to F-actin with dissociation constants of 0.6 and 12 microM, respectively. The 2-kDa peptide was found to compete with rabbit skeletal S-1 for the binding to F-actin by examining the binding of S-1 to actin in the presence of various concentrations of the 2-kDa peptide. The 2-kDa peptide was also shown to interact with the regulatory light chain of aorta myosin by difference UV absorption spectroscopy. The 2-kDa peptide inhibited the actin-activated ATPase activity of skeletal S-1, and the inhibition was canceled by the addition of the isolated regulatory light chain. These results suggest that the newly found 2-kDa peptide region may be related to the regulation of smooth muscle actomyosin ATPase activity by phosphorylation of the regulatory light chain. PMID- 8428913 TI - Quantitative characterization of the structure of rhodopsin in disc membrane by means of Fourier transform infrared spectroscopy. AB - Fourier transform infrared (FTIR) spectroscopy has been used for the detailed characterization and quantification of the secondary structure of bovine rhodopsin in native disc membranes. FTIR spectra were obtained in aqueous media, both in 1H2O and in 2H2O. Analysis of spectra by means of Fourier self deconvolution, complemented with maximum likelihood restoration and Fourier derivative, has allowed the characterization of major amide I secondary structure sensitive component bands of structural relevance which had not been detected before. In consequence, we show a richer secondary structure for rhodopsin than previously described. Our results indicate a total regular helix content around 51%, which would include not only the main alpha 1-type helix but also 3(10)-like helix. The presence of distorted helicoid sequences might furthermore increase to a certain extent the total helix amount. It is also indicated that a significant proportion of the amino acid residues are involved in extended/beta-structures and in reverse turns, as well as in "random" segments, which had not been directly demonstrated before. 61 +/- 4% of rhodopsin is determined to be solvent accessible, which is a substantially higher value than previously reported. Helices account for most of the inaccessible moiety. PMID- 8428914 TI - Enhanced stimulation of myosin subfragment 1 ATPase activity by addition of negatively charged residues to the yeast actin NH2 terminus. AB - We examined the effects of yeast actin NH2-terminal mutations on actomyosin interactions and the function of actin in vivo through measurements of actin activated ATPase activity, cosedimentation with rabbit muscle myosin subfragment 1 (S-1), in vitro motility, and invertase secretion assays. As reported earlier (Cook, R. K., Blake, W., and Rubenstein, P. A. (1992) J. Biol. Chem. 267, 9430 9436), elimination of NH2-terminal acidic residues from yeast actin results in an increased actin bundling, decreased actin-activated S-1 ATPase, and complete inhibition of actin filament sliding over myosin. Here we show that the addition of 2 new acidic residues to the NH2 terminus of yeast actin increased the Vmax value and the catalytic efficiency of the actin-activated ATPase activity of S-1. However, the binding of actin to S-1 in the presence of ATP and the velocities of actin sliding over myosin in the in vitro motility assays were not affected by this mutation. Thus, the number of actin NH2-terminal negative charges is important for actin activation of myosin S-1 ATPase activity, while only a minimum number of acidic residues is required for actin sliding over myosin in vitro. The number of actin NH2-terminal negative charges therefore appears to determine the efficiency with which the energy from ATP hydrolysis is converted to filament sliding. PMID- 8428915 TI - The x-ray crystal structure refinements of normal human transthyretin and the amyloidogenic Val-30-->Met variant to 1.7-A resolution. AB - The x-ray crystal structures of normal human transthyretin (prealbumin) and the amyloidogenic Val-30-Met variant have been refined at 1.7-A resolution to R values of 0.168 and 0.179, respectively, for 19,882 and 20,362 reflections (Fobs > 2.0 sigma). Standard deviations for stereochemical parameters are 0.018 and 0.022 A for bond distances, 0.030 and 0.038 A for angle distances, and 0.035 and 0.070 A for planar 1-4 distances. The newly refined normal structure shows improvement over the original structure of Blake and Swan (Blake, C. C. F., and Swan, I. D. A. (1971) J. Mol. Biol. 61, 217-224) in stereochemistry and in the conformation of the loop regions. Residues Arg-103, Thr-123, Asn-124, and Pro-125 have now been resolved, and residues 1-9 and 126-127 have been modeled with the aid of simulated annealing refinement. The functional form of transthyretin is a tetramer, having a cylindrical cavity which will bind thyroxine and an exterior binding site for the complex of retinol with retinol-binding protein. The monomer is a beta barrel flattened to become more like a sandwich with residue 30 in the interior. The methionyl for valyl substitution forces the beta sheets of the monomer as much as 1 A apart, resulting in a distortion of the thyroxine-binding cavity, in agreement with the independent observations that the Met-30 variant has low affinity for thyroxine. PMID- 8428916 TI - X-ray crystal structure of the Ala-109-->Thr variant of human transthyretin which produces euthyroid hyperthyroxinemia. AB - The structure of the Ala-109-->Thr mutation of human transthyretin, a nonamyloidogenic variant with enhanced thyroxine binding, has been determined by x-ray diffraction to a resolution of 1.7 A. The model, including 175 solvent water molecules, has been refined by constrained least squares to an R-value of 0.157. The standard deviations for protein geometry are 0.016 A for bond distances, 0.5 degree for bond angles, 0.031 A for 1-4 distances, and 0.005 A for deviations of planar groups from their least squares plane. The estimated error in protein atomic coordinates is 0.12 A. Residue 109 extends inward between the two beta sheets which form the major component of the monomer, as does the side chain of residue 30 in the amyloidogenic Met-30 variant. Comparison of the Thr 109 structure with that of the normal shows that the extra atoms of the threonine fit into empty space between sheets and make no extensive changes to the molecular conformation. The substitution at 109 causes small local changes in the secondary structure of the A, G, and H strands resulting in a shift of residues 15-17, 108-110, and 117 in each monomer. The thyroxine-binding sites of the Thr 109 and Met-30 variants and of the normal protein are compared, and the results suggest that the variation in affinity for thyroxine between the three proteins may arise from differences in the size of the binding pocket. PMID- 8428917 TI - NMR studies of phospholipase C hydrolysis of phosphatidylcholine in model membranes. AB - Hydrolysis of phospholipids in biological membranes by phospholipase C (PLC) produces an important second messenger molecule, 1,2-diacylglycerol (DAG), that is essential for the activation of protein kinase C (PKC). While the effects of DAG on model membranes have been investigated earlier, studies on physical properties of DAG introduced into phospholipid bilayers by PLC have been lacking. We present an NMR approach for studying structural and kinetic aspects of PLC mediated hydrolysis of 13 carbonyl-enriched phospholipids in model membranes (small unilamellar vesicles). The product DAG is readily detected by 13C NMR, and its structural properties as well as those of the model membrane can be monitored continuously. PLC hydrolysis was limited to a low proportion of the model membrane by incorporating a small amount of ester phospholipid into a nonhydrolyzable ether-linked phospholipid matrix. Under these conditions, PLC (Bacillus cereus) hydrolyzed only the monolayer of phosphatidylcholine to which it was exposed (the outer monolayer). The 1,2-DAG product remained associated with the membrane bilayer and did not alter bilayer structure in any detectable way. From the chemical shift data, it is inferred that the DAG has an interfacial conformation similar to that of phosphatidylcholine. These results show that DAG could activate PKC by direct interaction with the enzyme rather than by perturbation of the membrane bilayer. PMID- 8428918 TI - Novel activity of a yeast ligase deletion polypeptide. Evidence for GTP-dependent tRNA splicing. AB - Yeast tRNA ligase possesses multiple activities which are required for the joining of tRNA halves during the tRNA splicing process: cyclic phosphodiesterase, kinase, adenylylate synthetase, and ligase. A deletion polypeptide of a dihydrofolate reductase-ligase fusion protein, designated DAC, was previously shown to join tRNA halves although ATP-dependent kinase activity was not measurable in the assay used. We describe here a characterization of the mechanism of joining used by DAC and the structure of the tRNA product. DAC produces a joined tRNA and a splice junction with a structure identical to that produced by DAKC, the full-length dihydrofolate reductase-ligase fusion. Furthermore, DAC can use GTP as the sole cofactor in the joining reaction, in contrast to DAKC, which can only complete splicing in the presence of ATP. Both enzymes exhibit GTP-dependent kinase activity at 100-fold greater efficiency than with ATP. These results suggest that a potential function for the center domain of tRNA ligase (missing in DAC) is to provide structural integrity and aid in substrate interactions and specificity. They also support the hypothesis that ligase may prefer to use two different cofactors during tRNA splicing. PMID- 8428919 TI - Multiple nucleotide cofactor use by yeast ligase in tRNA splicing. Evidence for independent ATP- and GTP-binding sites. AB - We have examined multiple cofactor usage by yeast tRNA ligase in splicing in vitro. The ligase mechanism of action requires expenditure of two molar equivalents of nucleotide cofactor per mole of tRNA product. Recent evidence (Westaway, S.K., Belford, H.G., Apostol, B.L., Abelson, J., and Greer, C.L. (1993) J. Biol. Chem. 268, 2435-2443) demonstrated that the ligase-associated kinase activity is more efficient with GTP as cofactor than with ATP. Employing a ligase fusion construct with dihydrofolate reductase (Apostol, B.L., Westaway, S.K., Abelson, J., and Greer, C.L. (1991) J. Biol. Chem. 266, 7445-7455) for purposes of enzyme purification, we performed joining assays demonstrating that ATP and GTP are the most effective combination of cofactors. ATP was essential to the joining reaction, while UTP, CTP, or ATP replaced GTP inefficiently. Specific and functionally independent binding sites were confirmed for ATP and GTP by direct binding measurement. A third site was implicated in UTP- and CTP-ligase interactions. Comparison of binding constants with Kapp values determined for nucleotide-dependent joining suggested both that nucleotide triphosphate binding may be limiting in tRNA joining and that tRNA ligation occurs most efficiently using GTP for the kinase reaction and ATP as the adenylylate synthetase cofactor. PMID- 8428920 TI - The activity-controlling phosphorylation site is not the same in the four acidic ribosomal proteins from Saccharomyces cerevisiae. AB - By using site-directed mutagenesis and chemical analysis of phosphopeptides, a unique phosphorylation site has been shown at serine 73 in the amino acid sequence of the Saccharomyces cerevisiae acidic ribosomal protein YP1 beta (L44'). The mutation in this position prevents in vitro phosphorylation by protein kinases that modify the wild-type polypeptide. The unphosphorylatable mutated protein is unable to bind to the ribosomes and to rescue the growth deficiency of yeast strains in which the corresponding original gene is inactivated by gene disruption. Sequencing of tryptic phosphopeptides has shown that acidic proteins YP1 alpha and YP2 alpha (L44) are also phosphorylated at positions near the carboxyl end. These results contrast with the data indicating that in the highly homologous protein YP2 beta, phosphorylation takes place at serine 19, close to the amino terminus. The results show that phosphorylation is definitely required for the biological activity of these ribosomal proteins. However, the differences in the phosphorylation sites suggest that the effect of this modification is not the same in all of them, confirming the heterologous role of these peculiar ribosomal components. In fact, the different context of the modification sites in the four polypeptides suggests the existence of more than one protein kinase specific for this set of proteins. PMID- 8428921 TI - Generation of active myeloid and lymphoid granule serine proteases requires processing by the granule thiol protease dipeptidyl peptidase I. AB - The proenzyme activation peptides predicted from cDNAs encoding each of the granule serine proteases synthesized by cytotoxic lymphocytes and myeloid and mast cells are composed of 2 residues. The mechanism by which these amino terminal dipeptides are cleaved to generate the active enzymes has not been elucidated. The comparable distribution of dipeptidyl peptidase I (DPPI) and serine proteases and the ability of DPPI to hydrolyze relevant dipeptide sequences suggested a role for DPPI in the processing and activation of granule serine proteases. This study demonstrates that inhibition of DPPI activity is associated with impairment of the generation of granule serine protease activity in CD8(+) T cells, lymphokine-activated killer cells, P815 mastocytoma cells, and U-937 myeloid cells. Inhibition of DPPI resulted in impairment of the generation of cathepsin G enzymatic activity without reduction in the amount of immunoreactive cathepsin G produced. In U-937 cells pulsed with [3H]isoleucine, inhibition of DPPI activity was associated with the accumulation of the inactive proenzyme form of cathepsin G bearing an amino-terminal dipeptide extension to the isoleucine residue that normally occupies the amino terminus of the enzymatically active protein. These results indicate that DPPI plays a requisite role in the post-translational processing and activation of members of the family of granule serine proteases expressed in bone marrow-derived effector cells. PMID- 8428922 TI - A trisaccharide acceptor analog for N-acetylglucosaminyltransferase V which binds to the enzyme but sterically precludes the transfer reaction. AB - Development of inhibitors specific for the glycosyltransferases involved in the biosynthesis of asparagine-linked sugar chains has been undertaken in the hopes that these compounds may serve as tools to elucidate the roles of complex carbohydrates in biological recognition events. We report here the first example of a glycosyltransferase acceptor analog in which strategic replacement of a nonreacting hydroxyl group with a larger substituent produces a molecule which is recognized by the enzyme but does not react because of a steric block to the glycosyl transfer reaction. N-Acetylglucosaminyltransferase V catalyzes the transfer of GlcNAc from the sugar nucleotide donor UDP-GlcNAc to the 6-OH group of mannose in the synthetic trisaccharide acceptor beta GlcNAc(1-->2)alpha Man(1- >6)beta Glc-O(CH2)7CH3 (Km = 23 +/- 2 microM; Vmax = 116 +/- 3 pmol/h) to form the tetrasaccharide beta GlcNAc(1-->2)(beta GlcNAc(1-->6))alpha Man(1-->6)beta Glc-O(CH2)7CH3. The acceptor analog produced by replacement of the adjacent nonreacting 4-OH group of the mannose residue with an O-methyl group was not a substrate for the enzyme but was found to be a good competitive inhibitor of GlcNAc transferase V with Ki = 14 +/- 2 microM. To test the theory that it was the presence of the large methyl group which prevented the glycosyl transfer reaction the 4'-deoxygenated analog was synthesized. It was found to be a good substrate with Km = 74 +/- 6 microM and an almost 5-fold higher kcat (Vmax = 535 +/- 13 pmol/h). NMR data show no evidence of important conformational differences between the trisaccharide analogs, and kinetic experiments detected no differences for the binding of UDP-GlcNAc in their presence. The conclusion was therefore reached that the large methyl group introduced on O-4' sterically prevented the formation of product even though both potential substrates were bound by the enzyme. This "steric exclusion" strategy offers potential for the design of inhibitors for that class of glycosyltransferases in which the reactive hydroxyl group is also an essential recognition element. PMID- 8428923 TI - Initial interaction between fibrin and tissue plasminogen activator (t-PA). The Gly-Pro-Arg-Pro binding site on fibrin(ogen) is important for t-PA activity. AB - Gly-Pro-Arg-Pro (GPRP) is a potent inhibitor of fibrin polymerization. It also causes a concentration-dependent inhibition of the activation of plasminogen by t PA when soluble desAABB-fibrinogen (fibrin II) or fibrinogen are used as promoters of t-PA. Fibrinogen has a much weaker promoter activity than does fibrin II. Experiments were undertaken to explain the mechanism of action of GPRP on plasminogen activation. Kinetic data indicated that when GPRP was present in the assay system, the fibrin(ogen)-GPRP complex was ineffective as a t-PA promoter. Only the free forms of fibrin II or fibrinogen were promoters of t-PA. GPRP specifically eluted t-PA, which was previously bound to a fibrin-Sepharose column, and also inhibited t-PA binding to fibrin-Sepharose in a concentration dependent manner. These experiments were compared to those with other tetrapeptides: GHRP, GPGG, and GRGD. Only GHRP, which is known to bind more weakly to fibrin(ogen) than GPRP, had any effect. These results suggest that the GPRP binding site on the fibrin(ogen) molecule is important for t-PA activation and is a binding site for t-PA in the initial interaction between t-PA and fibrin. We propose that at or near the GPRP binding site on fibrin there is an initial binding site for t-PA. We hypothesize that kringle 1, which contains the sequence G128RRP, may be involved in the initial binding of t-PA to fibrin. PMID- 8428924 TI - Alternative substrates for calf intestinal adenosine deaminase. A pre-steady state kinetic analysis. AB - The protein fluorescence of adenosine deaminase (ADA) was perturbed during the deamination of adenosine and four analogues of adenosine. The kinetics for the approach to the steady-state during turnover were monitored by fluorescence changes associated with formation of enzymatic intermediates. These kinetic data and the steady-state kinetic data were analyzed in terms of the kinetic scheme as follows. [formula: see text] The steady-state turnover number was assigned to k2, which was 244 s-1 for adenosine and 1.1 x 10(-3) s-1 for 6-methylamino-2 aminopurine arabinoside (aMDAP). Values for the association rate constants, k1, and the dissociation rate constants, k-1, were calculated from the kinetics for the approach to the steady state. k1 varied from 31 x 10(6) M-1 s-1 for adenosine to 2.8 x 10(6) M-1 s-1 for N-6-methyladenine arabinoside. k-1 varied from 500 s-1 for adenosine to 31 s-1 for aMDAP. The latter value was confirmed (22 s-1) by spectrofluorometrically monitoring the trapping of ADA by excess erythro-9-(2 hydroxy-3-nonyl) adenine as aMDAP.ADA dissociated. The ratio of k2 to k-1, which determines the commitment to catalysis, decreased from 0.49 for adenosine to 3.5 x 10(-5) for aMDAP. The Km values calculated from k1, k-1, and k2 were similar to the values determined from steady-state kinetic data. The spectrum of enzyme bound aMDAP resembled protonated aMDAP. PMID- 8428925 TI - Deletion mutagenesis of high molecular weight kininogen light chain. Identification of two anionic surface binding subdomains. AB - The light chain (LC) of cleaved high molecular weight kininogen (HK) binds to anionic surfaces as well as the zymogens prekallikrein and factor XI and thus accelerates activation of the kallikrein-kinin, fibrinolytic, and coagulation pathways. The binding sites on HK LC for factor XI (amino acid residues 574-631) and prekallikrein (residues 583-613) have been localized to domain 6. Domain 5 (residues 438-520) has been postulated to contain the anionic surface binding subdomain. In order to define this subdomain we have expressed HK LC (residues Lys438-Ser644) as a fusion protein with glutathione-S-transferase (GST) in Escherichia coli and generated various HK LC deletion mutants. The recombinant HK LC (rHK LC) and various HK LC fragments were purified as GST fusion proteins by glutathione-Sepharose affinity chromatography from bacterial cell extracts. The rHK LC and recombinant fragments His459-Ser644, Glu466-Ser644, Leu483-Ser644, His493-Ser644, Lys438-Asp492, Lys438-Ser531, and His493-Lys520 inhibited 125I-HKa binding to kaolin, a model anionic surface used in the contact system, in a concentration-dependent manner. Deletion mutant proteins lacking domain 5, Thr521 Ser644 and Ser583-Ser644, did not inhibit the radiolabeled HKa binding to kaolin. The rHK LC and recombinant fragments Lys438-Asp492, Lys438-Ser531, His493-Ser644, His493-Lys520, Thr521-Ser644, and Ser583-Ser644 were radiolabeled with 125I and were then tested for their ability to bind to kaolin in the presence of fibrinogen and albumin. Except for the Thr521-Ser644 and Ser583-Ser644 fragments, all other radiolabeled HK LC deletion mutant proteins and rHK LC bound to kaolin in a concentration-dependent manner. This binding to kaolin was specific since it was inhibited by the addition of excess unlabeled HKa. The rHK LC, His493-Ser644 and delta 493-520 HK LC have coagulant activity, while other deletion mutant proteins did not exhibit coagulant properties. We conclude that there are at least two anionic surface binding subdomains, one in the histidine-glycine-rich region (Lys438-Asp492) and the other in the histidine-glycine-lysine-rich region (His493-Lys520), in the domain 5 of HK LC. Either subdomain, in the presence of the zymogen binding domain 6, is sufficient to impart coagulant activity to HK LC, while the presence of both did not increase the coagulant activity of HK LC additively. PMID- 8428926 TI - Contribution of specific cis-acting elements to activity of the mouse pro-alpha 2(I) collagen enhancer. AB - Intronic transcriptional regulatory regions are found in several collagen genes. We demonstrate here the importance of two distinct cis-acting elements for the activity of a transcriptional enhancer located in the first intron of the mouse pro-alpha 2(I) collagen gene. Enhancer subfragments were tested for their ability to stimulate a linked promoter following transient transfection into NIH/3T3 fibroblasts. A 92-base pair subfragment retained significant enhancer activity. DNase I footprinting identified a binding site for a nuclear factor (designated CIBF-I) within this subfragment. Electrophoretic mobility-shift experiments demonstrated that an oligonucleotide containing 18 base pairs from the protected region was sufficient for specific binding of CIBF-I and suggested that CIBF-I may be related to a protein(s) that binds to the rat c-mos enhancer. A second cis acting element, identified by electrophoretic mobility-shift and methylation interference experiments, bound affinity-purified Sp1 and an Sp1-like protein present in crude nuclear extracts. Small deletion mutations in either the CIBF-I or Sp1 site abolished factor binding in vitro and reduced enhancer activity in vivo. Our results represent the first identification of specific cis-acting elements required for full activity of the pro-alpha 2(I) collagen enhancer. PMID- 8428927 TI - Interactive intermediates are formed during the urea unfolding of rhodanese. AB - Structural transitions have been studied on the pathway for urea denaturation of rhodanese. Unlike guanidinium hydrochloride, urea gives no visible precipitation. Increasing urea concentrations cause a transition in which the enzyme activity is completely lost by 4.5 M urea, and there is a shift of the intrinsic fluorescence maximum from 335 nm for the native enzyme to 350 nm. There is a maximum exposure of organized hydrophobic surfaces at 4.5 M urea as reported by the fluorescence of 1,1'-bi(4-anilino)naphthalene-5,5'-disulfonic acid. Above 4.5 M urea, this probe reports the progressive loss of organized hydrophobic surfaces. The polarization of the intrinsic fluorescence falls with increasing urea concentrations in a complex transition showing that rhodanese flexibility increases in at least two phases. Rhodanese becomes increasingly susceptible to digestion by subtilisin between 3.5 and 4.5 M urea, giving rise to large fragments. At urea concentrations > 5 M, rhodanese is completely digested. There is a small increase in the rate of sulfhydryl accessibility between 3.5 and 4.5 M urea, but there is a large increase in the sulfhydryl accessibility above 4.5 M urea. Dimethyl suberimidate cross-linking shows the presence of associated species in 3-5 M urea, but there are few cross-linkable species at lower or higher urea concentrations. These results are consistent with a model in which urea unfolding of rhodanese is associated with the initial production of a species having organized regions of structure with exposed hydrophobic surfaces separated by flexible elements. PMID- 8428928 TI - Purification and structural characterization of transcriptional regulator Leu3 of yeast. AB - The transcriptional regulatory protein Leu3 of Saccharomyces cerevisiae was enriched approximately 70-fold above wild type level in yeast cells carrying a pGAL1-LEU3 expression vector. Sustained overproduction of Leu3 following induction by galactose required elevated intracellular levels of alpha isopropylmalate, a leucine pathway intermediate known to act as transcriptional co-activator. Starting with galactose-induced cells, the Leu3 protein was purified about 3,500-fold (i.e. 245,000-fold over wild type level) by a procedure that included treatment of the cell-free extract with polyethylenimine, fractionation with ammonium sulfate, heat treatment, and DNA affinity chromatography. Highly purified preparations still showed two protein bands when subjected to polyacrylamide electrophoresis under denaturing conditions. Their apparent molecular masses were about 104,000 and 110,000 kDa. The smaller of these values was very close to the maximum molecular weight obtained previously for Leu3 protein translated in vitro in a rabbit reticulocyte lysate. (The molecular weight deduced from the open reading frame of the LEU3 gene is 100,162.) Both protein bands reacted with antibodies raised against different portions of the Leu3 molecule and were, therefore, likely to represent two forms of Leu3. Treatment with calf intestinal phosphatase quantitatively converted the slower moving band into the faster moving one. Conversion was prevented by inorganic phosphate, a phosphatase inhibitor. These experiments showed that the two bands very likely correspond to phosphorylated and nonphosphorylated forms of Leu3. Phosphorylation did not appear to affect the DNA binding function of Leu3, but (indirect) effects on the activation function or effects on the modulation by alpha-isopropylmalate have not been ruled out. Electrophoretic mobility shift assays were used to estimate the apparent dissociation constants of the two specific Leu3-DNA complexes routinely seen in these assays. The values obtained were 1.1 and 2.6 nM. Finally, using size exclusion chromatography, native Leu3 protein was shown to have dimeric structure, irrespective of the state of phosphorylation. PMID- 8428929 TI - Independent binding of interleukin-1 alpha and interleukin-1 beta to type I and type II interleukin-1 receptors. AB - Interleukin (IL)-1 refers to a group of three polypeptide hormones with a wide range of cellular targets. Two types of IL-1 receptor have been identified and characterized by cDNA cloning. Both human type I and type II IL-1 receptors contain extracellular domains of approximately 310 residues and a single membrane spanning region. The type I receptor contains a cytoplasmic domain of 213 residues. The cytoplasmic region of the type II receptor is 29 residues in length. It has been found recently that a number of cells express both forms of receptor. By analogy with other cytokine receptor systems, the two IL-1 receptors might be expected to form a heterodimeric complex, the type II receptor being an alpha-chain-like structure, functioning only to bind ligand, and associating with the type I receptor (a beta-chain-like structure) which would transduce signals. In this report we show that this is not the case, but rather that IL-1, when complexed to type II receptor, cannot bind type I receptors, and vice versa. These data show that the complex patterns often observed for IL-1 binding to cells cannot be accounted for by the same type of mechanism that underlies the behavior of, for example, the IL-2 system. PMID- 8428930 TI - Analysis of transcriptional stimulation by recombinant Oct proteins in a cell free system. AB - The transactivation potential of several isoforms of the lymphoid-specific transcription factor Oct2 has been analyzed using in vitro transcription. Oct2 can stimulate transcription in B-cell nuclear extracts and in HeLa nuclear extracts depleted of the ubiquitous factor Oct1 by wheat germ lectin affinity chromatography. Activity is observed from both natural and synthetic promoters containing single or multiple copies of the octamer motif ATGCAAAT. Multimerization of this motif does not result in a synergistic transcriptional stimulation, but rather leads to a linear increase in activity. To analyze the various Oct2 isoforms, they were overexpressed in HeLa cells using recombinant vaccinia virus. Although all the isoforms bind similarly to the octamer sequence, they show clear differences in their ability to transactivate transcription. This ranges from a 2-fold stimulation for Oct2.3 to the almost 20-fold effect of the most potent variant Oct2.5. In general the relative activity of the isoforms in vitro reflects that observed in vivo in cotransfection experiments. Interestingly the ubiquitous factor Oct1 is also an efficient activator of transcription in vitro, but only from promoters with multiple octamer motifs. Sarkosyl inhibition studies suggest that both Oct1 and Oct2 function in vitro by stabilizing preinitiation complexes without affecting the reinitiation rate of RNA polymerase II. PMID- 8428931 TI - Iron content of human 5-lipoxygenase, effects of mutations regarding conserved histidine residues. AB - Recombinant human 5-lipoxygenase was expressed in Escherichia coli and purified to more than 95% homogeneity by ammonium sulfate precipitation and agarose-ATP column chromatography. The specific activity of the purified enzyme was 21-28 mumol/mg, as assessed by the generation of 5-hydro(pero)xyeicosatetraenoic acid. The iron content was analyzed by graphite furnace atomic absorption spectrophotometry for six preparations of the enzyme. The average value of the iron content was 0.86 mol/mol (iron/protein) with a range of 0.74-1.15 mol/mol. All lipoxygenases that have been sequenced contain 6 conserved histidine residues. Mutants of 5-lipoxygenase, with substitutions of these 6 conserved histidines, were purified and analyzed. Mutants H372Q and H550Q had no detectable enzyme activity and were also practically devoid of iron. Three mutants regarding His367 (H367Q, H367N, and H367S) were all inactive but had partial iron contents (0.5, 0.2, and 0.5 mol/mol, respectively). Finally, the mutated proteins H362Q, H390Q, and H399Q displayed reduced enzyme activity but contained similar amounts of iron as non-mutated 5-lipoxygenase. We conclude that histidines 372 and 550 constitute two of the iron ligands in 5-lipoxygenase. Also His367 is necessary for the enzyme activity, but this residue is not crucial for binding of iron. PMID- 8428932 TI - The novel metallothionein genes of Caenorhabditis elegans. Structural organization and inducible, cell-specific expression. AB - Two genes (mtl-1 and mtl-2) that encode the novel metallothioneins (MTs) of Caenorhabditis elegans (CeMTs) were cloned and characterized. Both genes contain a single intron that interrupts codon 6 and short 3'-untranslated regions. However, their promotor regions are distinctively non-homologous. The mtl-2 promoter contains a TATAA box and a single putative metal regulatory element. These elements are absent in the mtl-1 promoter. Nevertheless, both CeMT1 and CeMT2 mRNAs are induced by cadmium and contain precisely initiated, 5' untranslated sequences. The inducibility and cell type specificity of metallothionein gene expression were investigated in transgenic C. elegans that carry the lacZ (beta-galactosidase) reporter gene under the control of an mtl-1 or mtl-2 promoter sequence. Upon treatment of transgenic C. elegans with cadmium or heat stress, the mtl-2:lacZ fusion gene is abundantly and exclusively expressed in the intestinal cells of larvae and adult animals. Expression is not detected in the absence of metal or heat shock. In contrast, an mtl-1:lacZ construct is constitutively expressed in the pharynx and induced by cadmium and heat shock in the intestinal cells of C. elegans larvae. The metal-inducible expression of the mtl-1:lacZ gene is attenuated in adult transgenic nematodes. Thus, the activity of each mtl promoter is modulated by metals as well as developmental and environmental factors. PMID- 8428933 TI - Expression, characterization, and tissue distribution of a new cellular selenium dependent glutathione peroxidase, GSHPx-GI. AB - We have characterized a new selenium-dependent glutathione peroxidase, GSHPx-GI, by expressing a GSHPx-GI cDNA isolated from human hepatoma HepG2 cells in human mammary carcinoma MCF-7 cells, which have virtually undetectable expression of either the classical cellular enzyme, GSHPx-1, or GSHPx-GI at the protein level. One of the G418-resistant clones, neo-D1, expresses the transfected GSHPx-GI cDNA. This is based on 1) the presence of an additional GSHPx-GI DNA restriction fragment detected by Southern analysis; 2) the presence of a 1.9-kilobase (kb) GSHPx-GI mRNA in addition to the 1.0-kb endogenous mRNA by Northern analysis; and 3) the appearance of a 22-kDa 75Se-labeled protein which is absent in parental MCF-7 cells revealed by SDS-polyacrylamide gel electrophoresis. GSHPx-GI expressed in neo-D1 is a tetrameric protein localized in cytosol. GSHPx-GI does not cross-react with antisera against human GSHPx-1 or human plasma glutathione peroxidase (GSHPx-P). Similar substrate specificities are found for GSHPx-1 and GSHPx-GI; they both catalyze the reduction of H2O2, tert-butyl hydroperoxide, cumene hydroperoxide, and linoleic acid hydroperoxide with glutathione, but not of phosphatidylcholine hydroperoxide. GSHPx-GI mRNA was readily detected in human liver and colon, and occasionally in human breast samples, but not other human tissues including kidney, heart, lung, placenta, or uterus. In rodent tissues, GSHPx-GI mRNA is only detected in the gastrointestinal tract, and not in other tissues including liver. In fact, GSHPx-GI appears to be the major glutathione dependent peroxidase activity in rodent GI tract. This finding suggests that GSHPx-GI could play a major role in protecting mammals from the toxicity of ingested lipid hydroperoxides. In conclusion, we have demonstrated that GSHPx-GI is the fourth member in the selenium-dependent glutathione peroxidase family, in addition to GSHPx-1, GSHPx-P, and phospholipid hydroperoxide glutathione peroxidase (PHGPX). PMID- 8428934 TI - Tumor necrosis factor stimulates the synthesis and secretion of biologically active nerve growth factor in non-neuronal cells. AB - Tumor necrosis factor-alpha (TNF) markedly stimulates the synthesis and secretion of immunoreactive nerve growth factor (NGF) in quiescent mouse fibroblasts, which is a result of increase in the NGF mRNA level. NGF produced by TNF-treated fibroblasts has a molecular mass of 13 kDa on SDS-polyacrylamide gel electrophoresis, which is consistent in size with the subunit of mouse beta-NGF, and induces neurite outgrowth in paravertebral sympathetic neurons. Several peptide growth factors such as basic fibroblast growth factor (bFGF) and epidermal growth factor also stimulate NGF production in the cells, but not platelet-derived growth factor. The dose responses of TNF and bFGF to stimulate NGF production in the cells are, respectively, similar to those to induce cell proliferation. However, no correlation is observed between the ability of these growth factors to stimulate NGF production and that to induce cell proliferation. Thus, the stimulation of NGF production in the cells seems to be a specific activity of TNF and some other growth factors. TNF stimulates the synthesis and secretion of NGF also in other cells such as human glioblastoma cells. These findings suggest that TNF plays a role in regulating neuronal cell function through an indirect mechanism by which it stimulates NGF production in glial cells and fibroblasts. PMID- 8428935 TI - Chronic ethanol treatment increases expression of inhibitory G-proteins and reduces adenylylcyclase activity in the central nervous system of two lines of ethanol-sensitive mice. AB - The possibility that a 7-day period of ethanol exposure could regulate expression of specific GTP-binding regulatory proteins was investigated in two distinct brain regions from two different lines of ethanol-sensitive mice. Following ethanol treatment, plasma membranes were prepared from cerebellum and pons of short and long sleep mice. Studies of membranes were performed to assess hormone sensitive adenylylcyclase activity and to quantify expression of G-protein subunits. Immunoblot analysis showed that levels of Gi alpha(1) and Gi alpha(2) were markedly increased in cerebellar and pons membranes from ethanol-exposed mice compared to controls. Treatment of short sleep mice with ethanol enhanced ADP-ribosylation of both a 41- and a 39-40 kDa protein catalyzed by pertussis toxin. Ethanol did not alter expression of Gs alpha as assessed by immunoblot analysis, cholera toxin-dependent ADP-ribosylation, or by the ability of detergent extracted Gs alpha to reconstitute a functional adenylylcyclase in membranes from S49 cyc- murine lymphoma cells, a cell line which genetically lacks Gs alpha. Moreover, ethanol exposure did not influence levels of G(o) alpha or G beta 35-36 in either cerebellar or pons membranes. Cerebellar and pons membranes from ethanol-exposed short sleep mice demonstrated significantly less adenylylcyclase activity following stimulation with GTP, GTP gamma S, AlF, forskolin, and stimulatory ligands for three distinct receptors which couple to Gs alpha. Pretreatment of membranes with pertussis toxin reversed the ethanol induced inhibition in adenylylcyclase activity. These observations were not limited to one line of mice but were also documented in a second line of ethanol sensitive mice (e.g. long sleep). We conclude that ethanol exposure enhances expression of Gi alpha(1) and Gi alpha(2) in ethanol-sensitive mice, and is associated with decreased adenylylcyclase activity. Enhanced expression of Gi alpha(1) and Gi alpha(2) may contribute to impaired signal transduction in the central nervous system, and reduce the efficacy of neurotransmitters which signal through the adenylylcyclase system. PMID- 8428936 TI - Expression of the alpha-myosin heavy chain gene in the heart is regulated in part by an E-box-dependent mechanism. AB - Proximal regulatory element B (PRE-B), located from positions -318 to -284 in the alpha-myosin heavy chain (MHC) promoter, stimulated expression from an otherwise weak alpha-MHC promoter fragment in primary rat neonatal cardiomyocytes but not in the C2C12 myogenic cell line. PRE-B interacted with alpha-MHC binding factor 2 (BF-2), a protein found in nuclear extracts from several neonatal rat tissues and cell types including cardiomyocytes. BF-2 DNA binding activity was greatly reduced in adult versus neonatal tissues. Methylation interference footprints indicated that BF-2 bound to an element that included an E-box consensus sequence. Site-directed mutations in the BF-2-binding site, that abolish BF-2 binding, reduced expression from the full-length alpha-MHC promoter by 70%. A BF 2-like protein interacts within the HF-1a element of the myosin light chain-2 (MLC-2) promoter suggesting that one of the proteins that regulates the alpha-MHC and MLC-2 genes is identical or closely related. Analysis of binding by competition gel shift experiments indicated that both BF-2 and HF-1a are E-box binding proteins. The alpha-MHC and MLC-2 genes encode contractile proteins which are precursors of myosin. Regulation by the same transcription factor might indicate that the expression of alpha-MHC and MLC-2 is coordinately controlled. PMID- 8428937 TI - Purification and characterization of a membrane-bound protein carboxyl methyltransferase from rat kidney cortex. AB - Class II protein carboxyl methyltransferases (EC 2.1.1.77) are known to exist predominantly in a soluble form in all cells studied so far. These enzymes have been purified to homogeneity from the cytosols of many mammalian tissues but not from membranes. We describe here the purification to apparent homogeneity of a membrane-associated protein carboxyl methyltransferase from the brush border membrane of rat kidney. The enzyme was purified by fast protein liquid chromatography on Superdex 75 and Mono-Q and by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and consists of a single 27,300 polypeptide. The purified enzyme recognizes exogenous substrate proteins such as ovalbumin and gamma-globulins as well as synthetic peptides containing a L isoaspartyl residue but not a synthetic peptide containing a farnesylated C terminal cysteine (S-farnesyl-LARYKC). The Km for S-adenosyl-L-methionine with ovalbumin as the substrate is 1.5 microM and the purified enzyme is sensitive to inhibition by S-adenosyl-L-homocysteine (Ki = 0.3 microM). Peptide map obtained after Staphylococcus aureus V8 protease digestion of brush border membrane protein carboxyl methyltransferase showed a fragmentation pattern that was identical to that obtained for a soluble protein carboxyl methyltransferase purified according to the same procedure, indicating a high degree of homology. These results support the notion that class II protein carboxyl methyltransferases are not restricted to a cytosolic localization and show that the membrane-bound form of this enzyme shares many characteristics with known cytosolic protein carboxyl methyltransferases. PMID- 8428938 TI - Interaction of reconstituted high density lipoprotein discs containing human apolipoprotein A-I (ApoA-I) variants with murine adipocytes and macrophages. Evidence for reduced cholesterol efflux promotion by apoA-I(Pro165-->Arg). AB - Interaction of cells with both native and reconstituted high density lipoproteins (rHDL) containing apolipoprotein (apo) A-I mediates efflux of cellular cholesterol. To characterize structural requirements for this activity in apoA-I, six different genetic apoA-I variants and the corresponding normal allele products were isolated from heterozygous carriers, reconstituted with dimyristoylphosphatidylcholine (DMPC) into discoidal HDL and compared with regard to their ability to release intracellular cholesterol from murine adipocytes and peritoneal macrophages. In previous studies we determined the amino acid changes of these variants: Pro3-->Arg, Pro4-->Arg, Lys107-->0, Lys107-->Met, Pro165- >Arg, and Glu198-->Lys (von Eckardstein, A., Funke, H., Walter, M., Altland, K., Benninghoven, A., and Assmann, G. (1990) J. Biol. Chem. 265, 8610-8617) and demonstrated that all apoA-I variants except apoA-I(Lys107-->0) form discoidal HDL particles with phosphatidylcholine analogues indistinguishable from normal apoA-I (Jonas, A., von Eckardstein, A., Kezdy, K. E., Steinmetz, A., and Assmann, G. (1991) J. Lipid Res. 32, 95-106). In the present study, all apoA-I.DMPC complexes except those containing apoA-I(Pro165-->Arg) promoted cholesterol efflux as effectively as those containing normal apoA-I. Cholesterol efflux from adipocytes obtained after 180 min of incubation with apoA-I(Pro165-->Arg).DMPC complexes was decreased by 30% although this variant was bound to adipocytes with normal affinity. By contrast, apoA-I(Lys107-->Met).DMPC complexes were decreased in their binding affinity compared to normal apoA-I.DMPC complexes but normally promoted cholesterol efflux. Incubation of mouse peritoneal macrophages with apoA I(Pro165-->Arg).DMPC complexes did also result in a 30% decreased efflux of radiolabeled cholesterol if compared to rHDL with the normal allele product from the same donor. Together the observations suggest that the substitution of proline at residue 165 interferes with the formation of a structural domain in apoA-I that is crucial for cellular cholesterol efflux stimulation. Furthermore, our results suggest that binding of HDL to adipocytes and cholesterol efflux promotion by HDL requires different structural domains. PMID- 8428939 TI - In vivo DNA-protein interactions at the divergent mercury resistance (mer) promoters. I. Metalloregulatory protein MerR mutants. AB - Regulation of transcriptional initiation of the Tn21 mercury resistance (mer) operon occurs at the divergent promoter region lying between the structural genes (merTPCAD) and a regulatory gene (merR). During repression, both promoters are negatively regulated by MerR bound to a dyadic operator located between the -10 and -35 hexamers of PTPCAD. Upon Hg(II) induction, MerR activates transcription only from PTPCAD and continues to repress transcription from PR. Using in vivo dimethyl sulfate and KMnO4 footprinting of the merOP region of strains carrying wild-type MerR or MerR mutants, we have dissected the steps in MerR-mediated positive and negative regulation of the divergent mer promoters. The greater sensitivity of primer extension footprinting allowed the resolution of details previously undetectable in vivo. Two MerR mutants unable to bind merOP DNA allow RNA polymerase to form an open complex preferentially at PR. The intensity of the PR open complex is considerably less than that which occurs upon MerR-Hg(II) activation of PTPCAD and considerably greater than that which occurs at PTPCAD when MerR is deleted; this is the first in vivo estimate of the relative strengths of these two promoters. Although retaining the wild-type capacity to sequester RNA polymerase in the closed complex, the four MerR mutants defective in Hg(II) binding do not distort the dyad DNA or foster open complex formation when the inducer is added. Two activation-defective MerR mutants foster closed complex formation as well as wild-type, but they do not distort the dyad DNA or foster open complex formation when the inducer is added, although they are also able to bind Hg(II). Two semiconstitutive inducible MerR mutants differ from each other in that one (which lies nearer the COOH terminus) does distort the dyad center upon Hg(II) induction, whereas the other (which lies in near the center of merR) exhibits no dyad distortion upon induction. Paradoxically, despite their relatively strong semiconstitutive expression of mer-lac transcriptional fusions, neither of these mutants has a detectable open complex in the absence of added Hg(II). PMID- 8428940 TI - In vivo DNA-protein interactions at the divergent mercury resistance (mer) promoters. II. Repressor/activator (MerR)-RNA polymerase interaction with merOP mutants. AB - Transcription of the Tn21 mercury resistance (mer) operon is regulated by MerR which represses and activates the mer structural genes (merTPCAD) in the absence and presence of Hg(II), respectively. The promoter for the structural genes (PTPCAD) is divergently overlapped with the promoter for the regulatory gene (PR), and a dyadic operator lies between the -10 and -35 hexamers of PTPCAD. Using in vivo dimethyl sulfate and KMnO4 footprinting of mutant mer operator promoter (merOP) DNA to observe MerR and RNA polymerase-mediated interactions with the merOP region, we have identified three distinct domains within the palindromic mer operator. Dyad domain I consists of the outermost bases on the left arm of the operator palindrome whose alteration causes a shift, but apparently not a major loss, in occupancy by MerR, and no decrease in RNA polymerase occupancy. Mutants in dyad domain I are semiconstitutive but support additional Hg(II)-induced open complex formation at PTPCAD. Dyad domain II consists of the four highly conserved inner bases ( ... GTAC ... GTAC ... ) of the seven-base interrupted dyad, alteration of which severely modifies both MerR and RNA polymerase contacts in the promoter region. Mutants in domain II generally allow constitutive open complex formation at PR. One unusual mutant of this group retains most of the wild-type dyad's ability to repress both promoters but is unable to support activation at PTPCAD in response to Hg(II), indicating that MerR undergoes a conformational change and that the required base contacts for activation are different than those for repression. Dyad domain III is tentatively defined by a mutant in the outermost base of the right palindrome arm which is unaffected in either MerR or RNA polymerase occupancy, however, a second lesion within the PTPCAD -10 hexamer of this mutant limits effective open complex formation. Other mutations lying solely within the -10 RNA polymerase recognition hexamer of PTPCAD are similarly competent in both MerR and RNA polymerase binding, but inadequate for open complex formation. One such mutant also affects the overlapping -10 hexamer of PR and results in reduced occupancy by both MerR and RNA polymerase, likely as a result of inefficient transcriptional initiation of merR mRNA. Finally, mutations affecting the -35 hexamer of PTPCAD bind MerR but not RNA polymerase.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8428941 TI - Gene-specific DNA repair of interstrand cross-links induced by chemotherapeutic agents can be preferential. AB - The gene-specific formation and repair of interstrand cross-links (ICL) were measured in the dihydrofolate reductase (DHFR) gene in hamster cells. Cells were treated with two different chemotherapeutic agents, nitrogen mustard and cisplatin, and the frequency of cross-links was quantified in the active gene and in a downstream, inactive region. About 5% of total lesions induced by these agents were ICL. Whereas the frequencies of cross-links formed were similar in the gene and in the noncoding region after cisplatin treatment, there were more nitrogen mustard-induced cross-links in the inactive region than in the active gene. At low levels of cross-linking, we found preferential DNA repair in the active gene as compared to the inactive region. At higher levels of cross linking, there was no difference in repair rates between the gene and the noncoding region due to an increase in the repair efficiency in the inactive DNA. Implications of fine structural organization of cross-link repair are discussed. PMID- 8428942 TI - Domain structure of an N-ethylmaleimide-sensitive fusion protein involved in vesicular transport. AB - N-Ethylmaleimide-sensitive fusion protein (NSF) is an essential component for protein transport between Golgi cisternae. Sequence analysis and proteolytic dissection reveal that NSF contains two tandem "ATP domains," each containing the consensus sequence for the binding of nucleotide. When Escherichia coli-produced Chinese hamster ovary NSF is purified, it exhibits a low, but significant, ATPase activity. The ATPase activity of NSF is sensitive to N-ethylmaleimide and influenced by monoclonal antibodies against recombinant NSF. PMID- 8428943 TI - ras protein activity is essential for T-cell antigen receptor signal transduction. AB - In a Jurkat cell model of T-cell activation an interleukin-2 promoter/reporter gene construct was activated by antigen receptor agonism in combination with the lymphokine interleukin-1. Antigen receptor signals could be mimicked by suboptimal activation of protein kinase C (PKC) with phorbol esters in combination with calcium mobilization by an ionophore. In cotransfection experiments, oncogenic rats obviated the need for PKC stimulation but did not replace either the calcium signal or interleukin-1. Activated ras expression also replaced the requirement for PKC stimulation in activation of the T-cell transcription factor NF-AT. A dominant inhibitory ras mutant specifically blocked antigen receptor agonism, indicating that ras activity is required for antigen receptor signaling. In addition, an inhibitor of PKC blocked both activated ras and phorbol ester stimulation, suggesting a role for ras upstream of PKC. PMID- 8428944 TI - Transformation of the signal peptide/membrane anchor domain of a type II transmembrane protein into a cleavable signal peptide. AB - Rabbit neutral endopeptidase-24.11 is a type II transmembrane protein with a 27 amino acid residue positively charged NH2-terminal cytoplasmic domain, a 23-amino acid residue hydrophobic signal peptide/membrane anchor domain, and a large catalytic COOH-terminal domain exposed on the exoplasmic side of the membrane. In order to study the mechanism of membrane anchoring of neutral endopeptidase 24.11, we created mutants in which the cytoplasmic tail was deleted. Expression of these mutants in COS-1 cells resulted in the secretion of approximately 10-20% of the protein into the culture medium, due possibly to the cleavage of part or all of the signal peptide/membrane anchor domain by the rough endoplasmic reticulum signal peptidase. In a second set of mutants, a hydrophilic sequence (GSQNS) was inserted midway in the signal peptide/membrane anchor domain of neutral endopeptidase-24.11. When this hydrophilic sequence was introduced into the full-length neutral endopeptidase-24.11, approximately 20% of the enzyme activity was recovered in the culture medium. This proportion increased to 93% when the cytosolic tail was deleted. Sequencing of the [3H]tyrosine- or [3H]isoleucine-labeled secreted protein indicated that proteolysis, possibly by signal peptidase, occurred on the COOH-terminal side of the signal peptide/membrane anchor domain. We conclude that the efficient cleavage of the signal peptide/membrane anchor domain and secretion of the protein require both the deletion of the cytosolic domain and the presence of a hydrophilic sequence. PMID- 8428945 TI - Fidelity of phi 29 DNA polymerase. Comparison between protein-primed initiation and DNA polymerization. AB - Phi 29 DNA polymerase is able to catalyze two different synthetic reactions: protein-primed initiation and DNA polymerization. We have studied the fidelity of phi 29 DNA polymerase when carrying out these two reactions. Global fidelity was dissected into three steps: insertion discrimination, mismatch elongation, and proofreading. The insertion discrimination of phi 29 DNA polymerase in DNA polymerization ranged from 10(4) to 10(6). The efficiency of mismatch elongation was 10(5)-10(-6)-fold lower than that of a properly paired primer terminus. These factors indicate that DNA polymerization catalyzed by phi 29 DNA polymerase is a highly accurate process. Conversely, the insertion fidelity of protein-primed initiation was quite low, the insertion discrimination factor being about 10(2). Mismatch elongation discrimination was also rather low: mismatched terminal protein (TP).dNMP complexes were elongated from 2- to 6-fold more slowly than the correct TP.dNMP complex. Even more, the 3'-->5' exonuclease activity of phi 29 DNA polymerase was unable to act on the TP.dNMP initiation complex, precluding the possibility that a wrong dNMP covalently linked to TP could be excised and corrected. Therefore, protein-primed initiation can be predicted as a quite inaccurate reaction. The problem of maintaining the sequence at the DNA ends is discussed in the context of a recently described model for protein-primed initiation. PMID- 8428946 TI - Purification and crystallization of 2,3-dihydroxybiphenyl 1,2-dioxygenase. AB - 2,3-Dihydroxybiphenyl 1,2-dioxygenase, an enzyme of the biphenyl biodegradation pathway that cleaves the first of the aromatic rings, was purified to apparent homogeneity from Pseudomonas sp. strain LB400 that had been engineered to hyperexpress the bphC gene. The enzyme had a subunit molecular mass of 33.2 kDa as determined by SDS-polyacrylamide electrophoresis. Kinetic studies indicate a KM of 7 +/- 1 microM for 2,3-dihydroxybiphenyl. The enzyme is strongly inhibited by substrate (Kss = 300 +/- 10 microM). Catechol, 3-methylcatechol, and 4 methylcatechol were cleaved less efficiently and showed weaker substrate inhibition. 3,4-Dihydroxybiphenyl was not a substrate for the enzyme. Ammonium sulfate and polyethylene glycol 6000 were used as precipitants to obtain two different crystal forms. Crystals grown from ammonium sulfate and polyethylene glycol 6000 had space groups of P4(2)2(1)2 and I222, respectively. Electron microscopy indicates that the enzyme is an octamer (265 kDa) consisting of subunits arranged in two planar tetramers in a staggered conformation. PMID- 8428947 TI - Cell biological studies with monoclonal and polyclonal antibodies against human casein kinase II subunit beta demonstrate participation of the kinase in mitogenic signaling. AB - Casein kinase II (CKII) is a highly conserved ubiquitous serine/threonine kinase composed of two catalytically active (alpha and/or alpha') and two regulatory (beta) subunits. It has been suspected that, among numerous other cellular functions, CKII might play a role in the control of mitogenic signaling. To test for such a role and its mechanism in intact cells, monoclonal antibodies (mAbs) were generated against CKII beta using a recombinant protein containing amino acids 20-200 of human CKII beta. The CKII beta-specific mAb with the highest reactivity, mAb IVG6 (classified as IgG1 with kappa light chains), was purified to homogeneity. It recognized a CKII beta epitope comprising the amino acids 140 156, a basic and highly conserved region. In addition, polyclonal antibodies (pAbs) were raised and made monospecific by affinity purification. pAbs-mediated quantitative immunofluorescence microscopy of human IMR-90 fibroblasts and/or Western blots of cell fractions revealed (i) CKII beta was present in exponentially growing cells at a 2-3-fold higher level than in quiescent cells, (ii) CKII beta was localized predominantly in the nucleus of cells (3-15-fold cytoplasmic level depending on cellular state and assay used), and (iii) the nuclear/cytoplasmic ratio of CKII beta was higher by a factor of 2 in exponentially growing cells. Consequently, mitogenic stimulation of quiescent cells by fetal calf serum doubled the nuclear/cytoplasmic ratio of CKII beta. The increase occurred within the 1st h of stimulation. The translocation of CKII beta into the nucleus was inhibited when mAb IVG6 was injected into the cytoplasm at the time of mitogenic stimulation. This microinjection also significantly inhibited the cell proliferation. The data imply that cytoplasmic CKII participates in the transmission of mitogenic signals by translocation into the nucleus. PMID- 8428948 TI - Alternative splicing of exons encoding the calmodulin-binding domains and C termini of plasma membrane Ca(2+)-ATPase isoforms 1, 2, 3, and 4. AB - Rat plasma membrane Ca(2+)-ATPase (PMCA) mRNAs were examined by S1 nuclease protection (isoform 1) and polymerase chain reaction (isoforms 1, 2, 3, and 4) and the corresponding genes were analyzed to determine the tissue-specific splicing patterns involving exons encoding the calmodulin-binding domains and C termini. Splicing of PMCA1 involves a single 154-nucleotide exon that can be either included or excluded; when the exon is included four different splice donor sites, at positions 87, 114, 152, and 154, can be utilized. PMCA2 mRNAs are generated either by the inclusion of a 172-nucleotide exon, by the inclusion of both the 172-nucleotide exon and a 55-nucleotide exon, or by the exclusion of both exons. Four PMCA3 mRNAs arise by alternative splicing of a 154-nucleotide exon, in patterns that are analogous to those of PMCA1, and additional mRNAs are generated by the inclusion of a 68-nucleotide exon immediately before the 154 nucleotide exon. The simplest splicing pattern occurs in PMCA4, where a single 175-nucleotide exon is either included or excluded. The alternative mRNAs for each of the four genes are expressed in a tissue-specific manner and encode enzyme variants with different combinations of calmodulin-binding domains and C termini. PMID- 8428949 TI - AP-1 antagonizes thyroid hormone receptor action on the thyrotropin beta-subunit gene. AB - Thyrotropin-releasing hormone (TRH) stimulates and thyroid hormone (T3) inhibits transcription of the thyrotropin beta-subunit gene (TSH-beta). The first exon contains DNA sequences necessary for both responses and binds both AP-1 and thyroid hormone receptor (T3R). T3 did not inhibit TSH-beta gene expression in a T3R-deficient cell line. Transfection of a T3R expression vector, however, resulted in a 70% inhibition of expression by T3, which was abolished by cotransfection of c-jun and c-fos expression vectors. Mutations surrounding the transcription initiation site and DNA binding studies demonstrate both a functional and structural interaction between c-jun and T3R. Thus, TRH, acting through AP-1, may alter the set point and magnitude of thyroid hormone negative feedback of the TSH-beta gene through an interaction between AP-1 and T3R. Other regulatory pathways acting through AP-1 may alter thyroid hormone action in man. PMID- 8428950 TI - The primary structure of rat ribosomal protein L23a. The application of homology search to the identification of genes for mammalian and yeast ribosomal proteins and a correlation of rat and yeast ribosomal proteins. AB - The amino acid sequence of the rat 60 S ribosomal subunit protein L23a was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L23a has 156 amino acids and a molecular weight of 17,684. Hybridization of the L23a cDNA to digests of nuclear DNA suggests that there are 18-20 copies of the L23a gene. The mRNA for the protein is about 600 nucleotides in length. Rat L23a is related to the yeast Saccharomyces cerevisiae L25, to the archaebacterial Methanococcus vannielii L23, to eubacterial Escherichia coli L23, and to other members of the L23 family of ribosomal proteins. A novel application of a routine homology search procedure was employed to identify a nucleotide sequence that could be used to design an oligodeoxynucleotide probe to screen a library for a cDNA that encodes rat L23a; this same procedure uncovered a number of previously unidentified genes for yeast ribosomal proteins in the GenBank DNA data base. In a correlation of rat and yeast ribosomal proteins 48 pairs are shown to be related. PMID- 8428951 TI - Recruitment of vaccinia virus RNA polymerase to an early gene promoter by the viral early transcription factor. AB - Transcription of vaccinia virus early genes in vitro requires the virally encoded RNA polymerase and early transcription factor, VETF. VETF is a promoter-binding protein with DNA-dependent ATPase activity. We have investigated the functional role of VETF in transcription activation by analyzing the interaction between the RNA polymerase and promoter DNA. Using a gel shift assay, a novel protein-DNA complex was detected that required both RNA polymerase and VETF. The complex was suggested to be a transcription initiation complex by its ability to incorporate 32P-labeled nucleotides in combinations compatible with synthesis of a short RNA chain. Competition binding studies indicated that the RNA polymerase associated specifically with a viral early promoter. These experiments demonstrate that VETF activates transcription by directly recruiting the RNA polymerase to the promoter. Sedimentation analysis showed that VETF and RNA polymerase did not form a stable complex unless promoter DNA was present, indicating that protein-protein contacts are not the sole basis for initiation complex assembly. DNase I cleavage and methylation interference analyses revealed a hyperreactive site in the center of the promoter. Radiolabeling of RNA in the RNA polymerase-promoter complex did not occur when AMP-PNP (adenyl-5'-yl imidodiphosphate) was substituted for ATP, suggesting that ATP hydrolysis is required for the initiation of transcription. A model is proposed to account for these findings. PMID- 8428952 TI - Human plastin genes. Comparative gene structure, chromosome location, and differential expression in normal and neoplastic cells. AB - Plastins are a family of actin-binding proteins that are conserved throughout eukaryote evolution and expressed in most tissues of higher eukaryotes. In humans, two ubiquitous plastin isoforms (L and T) have been identified. The L isoform is expressed only in hemopoietic cell lineages, while the T isoform has been found in all other normal cells of solid tissues that have replicative potential (fibroblasts, endothelial cells, epithelial cells, melanocytes, etc.). However, L-plastin has been found in many types of malignant human cells of non hemopoietic origin suggesting that its expression is induced accompanying tumorigenesis in solid tissues. To learn more about the nature of plastin genes and their potential role in malignancy, the L- and T- plastin genes were cloned and sequenced to characterize their structure and mechanisms of regulation of expression. Each gene was found to be approximately 90 kilobases in size and was composed of 16 exons. All exon-intron junction sequences were identified and shown to conform to the canonical junction sequences. It was evident from their similar structure and coding homology that the two plastin genes have diverged from a common ancestor gene. L- and T-plastin genes were also mapped to chromosomes 13 and X, respectively, using polymerase chain reaction amplification with isoform-specific probes. An expanded survey of normal cell types and 50 tumor cell lines, demonstrated that 68% of carcinomas and 53% of other solid tumors of nonepithelial origin exhibited L-plastin expression, whereas the normal stem cell progenitors did not. Fibrosarcomas (n = 4), ovarian carcinomas (n = 9), breast carcinomas (n = 4), and choriocarcinomas (n = 2) combined exhibited the highest frequency and levels of L-plastin expression (95% frequency). In addition, 4 tumor cell lines that were L-plastin-negative exhibited evidence of defective T-plastin expression increasing the apparent co-incidence of plastin abnormalities associated with human tumorigenesis to 71%. Evidence is presented in support of a trans-activation mechanism for activation of L-plastin synthesis accompanying tumorigenesis. The induction of L-plastin expression accompanying SV40-mediated transformation of human embryonic lung MRC-5 fibroblasts was also confirmed. Finally, we present evidence that fimbrin is a third distinct plastin isoform which is specifically expressed at high levels in the small intestine. PMID- 8428953 TI - Characterization of the human L-plastin gene promoter in normal and neoplastic cells. AB - Plastins are a family of human actin-binding proteins (isoforms) which are abundantly expressed in all normal replicating mammalian cells. One isoform, L plastin, is constitutively expressed at high levels in hemopoietic cell types while T-plastin is constitutively expressed in all non-hemopoietic cells of solid tissues that have replicative potential (fibroblasts, endothelial cells, epithelial cells, melanocytes, etc.). L-plastin is, however, constitutively synthesized in many types of malignant human cells of solid tissues suggesting that its expression is induced during tumorigenesis. The frequency of L-plastin induction in some cancers of the steroid-regulated female reproductive tract (breast, ovary, uterus, and placenta) appears to be especially high (79% in a limited survey). To learn the mechanism of L-plastin gene activation accompanying tumorigenesis, we have begun to characterize the promoter and regulatory elements of the L-plastin gene. Transcription initiation from this promoter was found to occur at multiple sites and as near as 10 base pairs from the 3'-side of the TATAAA box. The promoter and its flanking DNA were cloned and sequenced to identify potential regulatory elements that participate in the induction of the L plastin gene in neoplastic cells. Examination of upstream sequences revealed the existence of two potential progesterone, one potential estrogen, and four potential Ets-1 responsive elements flanking the promoter. A 315-base pair fragment spanning the TATAAA box and a potential Sp1-binding site exhibited maximum promoter activity using CAT as a reporter while longer promoter fragments extending into upstream flanking sequences spanning the hormone receptor-response elements exhibited reduced promoter activity. An expression vector, pHLPPr-1-neo, was constructed using a 5.1-kilobase pair EcoRI-HindIII fragment of the L-plastin gene that contained the potential upstream regulatory elements, the TATAAA box, and part of the first exon. This promoter could direct the constitutive expression of the reporter beta-galactosidase at high frequency in transfected colonies of transformed cells that express L-plastin constitutively; by contrast, this promoter was virtually inactive in transfected colonies of normal fibroblasts and it exhibited a low frequency of constitutive activation in transfected colonies of in vitro SV40-transformed fibroblasts which did not exhibit L-plastin expression. The utility of this recombinant promoter in determining the mechanism(s) that leads to activation of the L-plastin gene in tumor cells is discussed. The potential significance of regulation of the L plastin gene by reproductive hormones in cancers arising in hormone-responsive tissues is also discussed. PMID- 8428954 TI - The specific subcellular localization of two isoforms of cytochrome b5 suggests novel targeting pathways. AB - Two forms of cytochrome b5 are present in rat tissues, with a sequence identity of approximately 60% in the cytoplasmically exposed, tryptic fragments (Lederer, F., Ghrir, R., Guiard, B., Cortial, S., and Ito, A. (1983) Eur. J. Biochem. 132, 95-102). It has been suggested that the two isoforms have partially overlapping subcellular distributions, with each form localized to some extent on both endoplasmic reticulum and outer mitochondrial membranes (Ito, A. (1980) J. Biochem. (Tokyo) 87, 73-80). To investigate the degree of specificity of the localization of cytochrome b5 isoforms, we studied their subcellular distributions with antipeptide antibodies, one specific for microsomal cytochrome b5, one specific for outer membrane cytochrome b, and one against a sequence common to the two cytochromes. We first identified outer membrane Cyt b as a tightly bound, Triton X-114-extractable, 23-kDa polypeptide. We then analyzed biochemically characterized rat liver subcellular fractions by Western blotting and found that outer mitochondrial membrane cytochrome b was not present on endoplasmic reticulum membranes. Conversely, microsomal cytochrome b5 was present on outer mitochondrial membranes in extremely low concentration, at a level < 5% of that on endoplasmic reticulum membranes. Thus, the subcellular distribution of microsomal cytochrome b5 is more restricted than previously thought, suggesting that novel posttranslational targeting mechanisms direct it to the endoplasmic reticulum. PMID- 8428955 TI - Identification of regions in the Ca(2+)-ATPase of sarcoplasmic reticulum that affect functional association with phospholamban. AB - When the SERCA 2 isoform of the Ca(2+)-ATPase of cardiac and slow-twitch muscle sarcoplasmic reticulum was coexpressed with phospholamban in COS-1 cells, a reduction in Ca2+ affinity (measured as Ca2+ dependence of Ca2+ transport) of 0.2 0.3 pCa units was observed. This inhibitory effect was reversed by phosphorylation of phospholamban with cAMP-dependent protein kinase A. SERCA 1 and SERCA 3, were also expressed in COS-1 cells, alone and together with phospholamban. SERCA 1 had high Ca2+ affinity which was reduced upon coexpression with phospholamban, but SERCA 3 had lower Ca2+ affinity, which was unaltered by coexpression with phospholamban. To identify which regions of the Ca2+ ATPase sequence determine its functional interaction with phospholamban, chimeric Ca(2+) ATPases between SERCA 2 and SERCA 3 were constructed and coexpressed with phospholamban. Measurement of Ca2+ affinities for a series of chimeras showed that two separate regions of the cytoplasmic domain of SERCA 2 were required for manifestation of a functional interaction between phospholamban and the Ca(2+) ATPase. The first is a region between amino acids 336 and 412 in the phosphorylation domain, which corresponds to a phospholamban interaction site identified earlier (James, P., Inui, M., Tada, M., Chiesi, M., and Carafoli, E. (1989) Nature 342, 90-92). The second region is the nucleotide binding/hinge domain (amino acids 467-762) which determines high Ca2+ affinity for SERCA type pumps (Toyofuku, T., Kurzydlowski, K., Lytton, J., and MacLennan, D. H. (1992) J. Biol. Chem. 267, 14490-14496). PMID- 8428956 TI - Purification and characterization of a protein tyrosine phosphatase containing SH2 domains. AB - A protein tyrosine phosphatase (PTP) containing two SH2 domains (PTP1C) was purified to near homogeneity from an adenovirus expression system by a two-step chromatographic procedure with a yield of 67%. The purified enzyme behaves as a monomer of 68 kDa on gel filtration and is totally specific for phosphotyrosyl residues. Its optimal pH is around neutrality for protein substrates such as reduced, carboxyamidomethylated, maleylated (RCM)-lysozyme and myelin basic protein but below 5 for low molecular weight compounds such as para-nitrophenyl phosphate (p-NPP) and phosphotyrosine. Furthermore, with the protein substrates, it displays an activity less than 1% of that obtained with other known PTPs but comparable activities toward p-NPP and phosphotyrosine. Its responsiveness toward the usual PTP activators (e.g. spermine) or inhibitors (e.g. vanadate, molybdate, heparin, or Zn2+) varied considerably with the nature of the substrates involved. Limited digestion with trypsin caused the cleavage of a C-terminal segment of the enzyme, giving rise to a 63-kDa fragment; this cleavage resulted in an approximately 20- and 10-fold activation of the enzyme toward RCM-lysozyme and myelin basic protein, respectively. PMID- 8428957 TI - Sarcina ventriculi synthesizes very long chain dicarboxylic acids in response to different forms of environmental stress. AB - Changes in the composition of membrane lipids in a strictly anaerobic, facultative acidophilic eubacterium, Sarcina ventriculi, were studied in response to various forms of environmental stress. Changes in lipid composition and structure occurred in response to changes in environmental pH. At neutral pH, the predominant membrane fatty acids ranged in chain length from C14 to C18. However, when cells were grown at pH 3.0, a family of unique very long chain fatty acids containing 32-36 carbon atoms was synthesized and accounted for 50% of the total membrane fatty acids. These acids were identified as very long chain alpha,omega dicarboxylic acids ranging in length from 28 to 36 carbons by electron impact mass spectrometry of methyl and (perdeuterio) methyl ester derivatives. These methyl esters all bore a vicinal dimethyl group toward the center of the chain. The assignment of the structures was confirmed by isolating one of the very long chain unusual fatty acids as the ester form after methanolysis and performing further analyses including 1H and 13C NMR spectroscopy and Fourier transform infrared spectroscopy. Coupling this information with the data from gas chromatography/mass spectrometry analysis, the exact structure was confirmed as alpha,omega-15,16-dimethyltricotanedioate dimethyl ester. Addition of alcohols, either metabolic (0.25 M ethanol) or nonmetabolic (0.05 M butanol) to cells grown at pH 7.0, or thermal stress (growth temperature at pH 7.0 was raised from 37 to 45 or 55 degrees C) also resulted in the synthesis of these very long chain fatty acids. Synthesis of these very long chain alpha,omega-dicarboxylic acids was reversed by reducing the temperature back to 37 degrees C. S. ventriculi is also unusual in that the membrane components are not the usual phospholipid components but appear to be predominantly glycolipids. PMID- 8428958 TI - Molecular characterization of the family of the N-methyl-D-aspartate receptor subunits. AB - cDNA clones for four different N-methyl-D-aspartate (NMDA) receptor subunits (NMDAR2A-NMDAR2D) were isolated through polymerase chain reactions followed by molecular screening of a rat brain cDNA library. These subunits are only about 15% identical with the key subunit of the NMDA receptor (NMDAR1) but are highly homologous (approximately 50% homology) with one another. They also commonly possess large hydrophilic domains at both amino- and carboxyl-terminal sides of the four putative transmembrane segments. NMDAR2A and NMDAR2C expressed individually in Xenopus oocytes showed no electrophysiological response to agonists. However, these subunits in combined expression with NMDAR1 markedly potentiated the NMDAR1 activity and produced functional variability in the affinity of agonists, the effectiveness of antagonists, and the sensitivity to Mg2+ blockade. Thus, NMDAR1 is essential for the function of the NMDA receptor, and multiple NMDAR2 subunits potentiate and differentiate the function of the NMDA receptor by forming different heteromeric configurations with NMDAR1. Northern blotting and in situ hybridization analyses revealed that the expressions of individual mRNAs for the NMDAR2 subunits overlap in some brain regions but are also specialized in many other regions. This investigation demonstrates the anatomical and functional differences of the NMDAR2 subunits, which provide the molecular basis for the functional diversity of the NMDA receptor. PMID- 8428959 TI - Biosynthesis of the vasoactive lipid monobutyrin. Central role of diacylglycerol. AB - Monobutyrin (1-butyrylglycerol) is a simple lipid secreted by adipocytes that stimulates both angiogenesis and vasodilation of microvascular beds. While monobutyrin production is increased during lipolysis, little is known about the biosynthetic pathway of this short chain monoacylglycerol. We show here that diacylglycerol, an intermediate in the lipolytic pathway, can stimulate monobutyrin synthesis from an adipocyte particulate fraction and does so by acting as a substrate. The biosynthetic route involves the acylation of diacylglycerol with butyryl-CoA to form a transient triacylglycerol intermediate. This intermediate is then hydrolyzed by a lipase(s) with specificity for long chain fatty acyl groups to form monobutyrin. Based on this information, a novel pathway for the synthesis of monobutyrin is proposed. Preliminary characterization of these enzymatic activities with respect to their cell-type specificity indicates that the acyltransferase activity is found in several cell types, albeit at lower levels than in adipocytes. The lipase activity appears to be two distinct steps, with both steps being adipocyte differentiation-dependent. These results indicate the likely biochemical basis for the restriction of monobutyrin synthesis to fat cells and also suggests a molecular basis for the close linkage between lipolysis, monobutyrin synthesis, and vasodilation. PMID- 8428960 TI - Novel secretory heparin-binding factors from human glioma cells (glia-activating factors) involved in glial cell growth. Purification and biological properties. AB - Growth factors for rat primary glial cells were identified in conditioned medium of a human glioma-derived cell line. The factors, designated glia-activating factors (GAFs), were purified to homogeneity by a combination of heparin affinity chromatography, gel filtration, and high performance liquid chromatography on a heparin affinity column and a C4 reversed-phase column. GAFs could be resolved into three peaks by C4 column chromatography. The M(r) values of these three proteins were estimated to be 30,000, 29,000, and 25,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. These M(r) values were in good agreement with the value of 26,000 +/- 3,000 estimated from the elution volume upon gel filtration chromatography under nondenaturing conditions. These data suggested that each of the GAFs consists of a single polypeptide chain and has no subunit structures. These three purified GAFs had almost the same growth-stimulating effect on glial cells in vitro, and the half maximal dose was around 10(-11) M. Concanavalin A staining and glycopeptide N glycosidase treatment of GAFs indicated that an asparagine-linked oligosaccharide chain(s) was attached to these three kinds of GAFs. Microsequencing of each GAF revealed a single amino-terminal sequence with no significant homology to any known protein, and the amino-terminal sequence of the 30-kDa GAF included that of the 29-kDa GAF. GAFs also stimulated the cell growth of oligodendrocyte type 2 astrocyte progenitor cells, BALB/c3T3 fibroblasts, and PC-12 cells but not that of human umbilical vein endothelial cells. PMID- 8428961 TI - Pancreatic-type phospholipase A2 stimulates prostaglandin synthesis in mouse osteoblastic cells (MC3T3-E1) via a specific binding site. AB - Previously, we have reported a novel proliferative action of pancreatic group I phospholipase A2 (PLA2-I) via a specific binding site in Swiss 3T3 fibroblasts, vascular smooth muscle cells, and chondrocytes. In this study, we characterized the PLA2-I specific binding site in osteoblastic cell line (MC3T3-E1 cells) with an equilibrium binding constant (Kd) value of 1.13 nM and maximum binding capacity of 40.1 fmol/10(6) cells. PLA2-I stimulated prostaglandin E2 (PGE2) production in a concentration-dependent manner in MC3T3-E1 cells, and its EC50 value was similar to the Kd value for PLA2-I binding. This effect of PLA2-I was type-specific and did not depend on its hydrolytic activity. PLA2-I increased the activity of prostaglandin endoperoxide synthase (PES), and PLA2-I-stimulated PGE2 synthesis was inhibited by cycloheximide. Northern blot analysis showed the increase in both type-1 and type-2 PES mRNAs. These findings indicated that PLA2 I stimulated PGE2 synthesis by induction of PES via a specific binding site in osteoblastic cells. PMID- 8428962 TI - Binding of protein S to factor Va associated with inhibition of prothrombinase that is independent of activated protein C. AB - Since plasma protein S serves an anticoagulant function by mechanisms which are not completely understood, its possible interaction with Factor Va was investigated. Human protein S bound to immobilized human Factor Va in a calcium dependent, saturable, and reversible manner and Factor Va bound similarly to immobilized protein S. Binding of protein S to immobilized Factor V was greatly enhanced by pretreatment of the surface-bound Factor V with increasing doses of thrombin up to 1 unit/ml. Binding of protein S to Factor Va was also demonstrated in fluid phase with a Kd of 33 +/- 9 nM. Biotin-labeled heavy chain of Factor Va bound to immobilized protein S, and this binding was reversed by a 17-fold molar excess of intact unlabeled Factor Va. Protein S competed efficiently with prothrombin for binding to immobilized Factor Va. The prothrombinase activity in a reaction mixture of purified clotting factors was inhibited by protein S and exhibited a pattern of mixed inhibition. The concentration of protein S needed for 50% inhibition of the prothrombinase activity of a mixture containing 1 nM Factor Xa, 20 pM Factor Va, and 50 microM phospholipids was about 16 nM. Since not all protein S preparations exhibited this degree of prothrombinase inhibitory activity, extensive control experiments were performed to verify that the inhibitory activity was associated with protein S during immunoaffinity chromatography and was not caused by traces of activated protein C in the protein S preparations. These data show that protein S has an anticoagulant function which is independent of activated protein C and, at least in part, that this is because of its competition with prothrombin for direct binding to Factor Va. PMID- 8428963 TI - High level thrombospondin 1 expression in two NIH 3T3 cloned lines confers serum- and anchorage-independent growth. AB - Thrombospondin (TSP) is a trimeric molecule synthesized by a variety of normal and transformed cells and secreted into the extracellular matrix. A number of studies have shown TSP to be intimately involved in the regulation of cellular proliferation. These include the findings that TSP is a mitogen-inducible immediate-early response gene; it localizes to areas of cellular proliferation in the developing mouse embryo; it augments the proliferative response to epidermal growth factor; anti-TSP monoclonal antibodies block cell proliferation; and TSP levels correlate directly with the growth and invasive phenotype and inversely with the degree of differentiation of squamous carcinoma cells. To determine whether TSP could behave as an oncogene, conferring serum and anchorage independence, NIH 3T3 cells were stably transfected with a TSP expression vector. Clones producing high levels of TSP displayed enhanced viability and a proliferative advantage in serum-reduced media. Moreover, this growth advantage could be specifically negated by treatment of the transfected cells with anti-TSP monoclonal antibodies. While the TSP overexpressing clones were capable of anchorage-independent growth in soft agar, surprisingly, they were incapable of forming tumors in nude mice, possibly due to the in vivo antiangiogenic activity of TSP. Regardless, these studies demonstrate that overexpression of thrombospondin, a constituent of the extracellular matrix, results in serum and anchorage independent growth, attributes normally associated with the transformed phenotype. PMID- 8428964 TI - A novel short consensus repeat-containing molecule is related to human complement factor H. AB - We have identified a novel factor H-related cDNA, which was isolated from a human liver cDNA library. The DOWN16 clone is 1269 base pairs in size and hybridized to a mRNA of 1.4 kilobases. Similar to the previously described factor H-related proteins, the predicted translation product of 331 amino acids contains a hydrophobic signal sequence followed by a stretch of five short consensus repeats (SCRs). These five SCRs display homology to SCRs of factor H: SCRs1-3 (DOWN16) are homologous to SCRs6-8 of factor H, while SCRs4 and -5 are related to SCRs19 and -20. In vitro translation demonstrated that the DOWN16 cDNA encodes a primary translation product of an apparent molecular mass of 37,500 Da which is directed to the secretory pathway and is glycosylated. Thus, we propose that the protein will be present in human serum. The relatedness of structural elements between this novel gene and factor H may suggest common functions of these proteins not yet determined. PMID- 8428965 TI - A specific CCAAT-binding protein, CBP/tk, may be involved in the regulation of thymidine kinase gene expression in human IMR-90 diploid fibroblasts during senescence. AB - The aging of IMR-90 human diploid fibroblasts in culture is accompanied by a 5-7 fold decrease in the level of thymidine kinase (TK) mRNA and TK activity (Chang, Z. F., and Chen, K. Y. (1988) J. Biol. Chem. 263, 11431-11435). We have employed a gel mobility shift analysis to investigate the molecular basis of the age dependent attenuation of TK gene expression. Several cis-elements including two inverted CCAAT boxes, located at base pairs (bp) -36 and -67, and GC-rich Sp1 binding sites have been identified in the TK promoter. A 28-bp (-91 to -64) fragment containing the distal inverted CCAAT element was excised from the TK promoter to examine possible differences in nuclear protein binding between young and old IMR-90 cells. A prominent DNA-protein complex was identified in serum stimulated young cells by a gel mobility shift assay. Competition analysis indicated that the binding was highly specific. The nuclear protein responsible for the complex formation was named CBP/tk (CCAAT Binding Protein for TK gene) since methylation interference assay showed that the inverted CCAAT box was involved in binding. The appearance of the CBP/tk-28-bp complex in IMR-90 cells was (i) serum-dependent, becoming prominent 12-24 h after serum stimulation, and (ii) age-dependent, prominent only in young but not in old IMR-90 cells. Similar serum- and age-dependent complex formations were also observed using a 67-bp fragment (-63 to +4) containing the proximal CCAAT element and a TATA box. In contrast, the binding activities for the Sp1 sequence were the same in young and old cells and appeared to be serum-independent. CBP/tk binding activity in nuclear extracts was abolished by heat (60 degrees C, 5 min) or treatment with proteinase K (0.1 microgram/ml) and sodium dodecyl sulfate (0.005%), but not by Nonidet P-40 or Triton X-100. Treatment of nuclear extracts with alkaline phosphatase or lectins (concanavalin A and wheat germ agglutinin) did not affect the binding activity. Metal chelators such as 1,10-ortho-phenanthroline (0.5 mM) inhibited the CBP/tk binding activity. Cycloheximide added to the serum stimulated cultures at an early or mid-G1 phase inhibited the CBP/tk binding activity. The half-life of the serum-induced CBP/tk binding activity was estimated to be less than 1 h.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8428966 TI - Characterization of the nuclear and cytoplasmic components of the lymphoid specific nuclear factor of activated T cells (NF-AT) complex. AB - The lymphoid-specific transcription complex, NF-AT, is involved in early gene activation in T cells and is assembled from a pre-existing, T cell restricted cytoplasmic factor and an inducible ubiquitous nuclear component within 30 min after activation through the antigen receptor. Recent studies have implicated the family of AP1 factors as components of the murine NF-AT complex. Evidence is provided here that the nuclear component of human NF-AT contains the phorbol ester-inducible transcription factor AP1 (Jun/Fos). We further characterize which AP1 family members can assume this role. Antisera to Fos inhibits NF-AT DNA binding as does an oligonucleotide containing a binding site for AP1. Constitutive expression in vivo of Fos, and to a lesser extent Fra-1, eliminates the requirement for phorbol 12-myristate 13-acetate (PMA) stimulation, leaving NF AT-directed transcription responsive to calcium ionophore alone. Overexpression of cJun or JunD, but not JunB, also eliminates the requirement for PMA, indicating that many but not all Jun- and Fos-related proteins functionally activate NF-AT-dependent transcription in the presence of the cytoplasmic component. NF-AT DNA binding can be reconstituted in vitro using semi-purified AP1 proteins mixed with cytosol from T lymphocytes. Fos proteins are not needed for this reconstitution, and although JunB is not functional, it can participate in the NF-AT DNA binding complex. Finally, we have partially purified the cytoplasmic component of NF-AT and show by elution and renaturation from SDS polyacrylamide gel electrophoresis gels that it has a molecular mass between 94 and 116 kDa and may have multiple differentially modified forms. PMID- 8428967 TI - Linkage of human spermatid-specific basic nuclear protein genes. Definition and evolution of the P1-->P2-->TP2 locus. AB - Protamines and transition proteins are highly basic sperm-specific nuclear proteins that serve to compact the DNA within the condensing spermatid nucleus. It has been proposed that the genes for protamine P1 and P2 and transition protein 2 may be clustered at a single loci. This may have marked implications for their evolution and coordinate developmental regulation. The cloning of a contiguous segment encompassing this region would directly address these issues. We have utilized a cloned segment of the human protamine P1 gene to isolate a human cosmid clone which contains the genes for protamines P1 and P2 and transition protein TP2. Sequence and hybridization analysis of this cosmid showed that the TP2 gene resided 3' of the P2 gene at a distance of approximately 7 kilobases (kb). This establishes the order of this gene cluster within a 13-kb region of 16p13.13-16p13.2 to be P1-->P2-->TP2. Similar to their spatial order, comparative computer-assisted sequence analysis predicts an identical relative rank order of emergence. Accordingly we propose that this gene cluster may have arisen from the P1 gene through a series of gene duplication events. PMID- 8428968 TI - Mammalian poly(A)-binding protein II. Physical properties and binding to polynucleotides. AB - The 49-kDa poly(A)-binding protein II (PAB II) was purified to homogeneity from calf thymus. The 70-kDa poly(A)-binding protein I (PAB I) was obtained in different fractions of the same preparation. Whereas PAB II stimulated poly(A) polymerase, PAB I was an inhibitor. In analytical ultracentrifugation, the predominant form of PAB II was a monomer of 50.3 kDa. A sedimentation constant of only 2.2 S indicated a distinctly non-spherical shape. Binding was specific for single-stranded purine polyribonucleotides. The dependence of the dissociation constant on the length of oligoriboadenylate indicated a binding site size of 12 nucleotides. A single site was bound with a KD of 2 x 10(-9) M, as determined by nitrocellulose filter binding assays. From fluorescence quenching and gel retardation experiments, the packing ratio on poly(A) was estimated as 23 nucleotides/protein monomer. PMID- 8428969 TI - Metabolism-based transformation of myoglobin to an oxidase by BrCCl3 and molecular modeling of the oxidase form. AB - The stoichiometric reductive debromination of BrCCl3 to a trichloromethyl radical by myoglobin caused the prosthetic heme to become covalently cross-linked to the protein moiety and transformed myoglobin from an oxygen storage protein to an oxidase. This was shown in experiments in which oxygen consumption was measured during redox cycling of the altered myoglobin in the presence of ascorbate or an enzymatic reducing system containing diaphorase and NADH. Redox cycling eventually led to loss of the protein-bound heme adduct and oxidase activity of myoglobin. We have used molecular modeling and the known structure of the protein bound heme adduct to identify probable mechanisms for transformation of myoglobin to an oxidase. Based on these modeling studies, the most likely structure of the experimentally observed adduct involves ligation to the heme iron of the epsilon nitrogen atom of histidine 97 and/or that of histidine 64. The model structures revealed access of solvent to the heme active site, which could facilitate oxygen reduction. The transformation of myoglobins and perhaps other hemoproteins to oxidases may have toxicological importance in causing the tissue damage resulting from exposure to various xenobiotics and endogenous chemicals as well as explaining how hemoproteins are inactivated during catalysis. PMID- 8428970 TI - Purification of a 90-kDa protein (Band VII) from cardiac sarcoplasmic reticulum. Identification as calnexin and localization of casein kinase II phosphorylation sites. AB - Purification and sequencing of proteins from cardiac sarcoplasmic reticulum (SR) vesicles have provided a framework for the study of SR function. Included among the SR proteins so far investigated are a collection of intralumenal proteins that stain blue with the protein dye Stains-All in sodium dodecyl sulfate polyacrylamide gels. A single prominent blue staining SR protein of apparent M(r) = 90,000 (Band VII), however, has remained uncharacterized. In the work described here, purification of Band VII from dog cardiac SR vesicles, along with amino acid sequencing and cDNA cloning, identified this protein as calnexin, a homologue of calreticulin recently described in dog pancreatic microsomes. Using ryanodine-mediated calcium oxalate loading of SR vesicles followed by density gradient centrifugation, we have shown that calnexin is a bona fide SR protein and an integral constituent of both junctional and free SR vesicles. Calnexin was found to be a substrate for casein kinase II and was phosphorylated at two distinct sites localized to the carboxyl- and amino-terminal ends of the molecule. Enrichment of calnexin in cardiac SR vesicles indicates a role for calnexin involving the specialized function of the SR membrane. PMID- 8428971 TI - Genomic organization of human surfactant protein D (SP-D). SP-D is encoded on chromosome 10q22.2-23.1. AB - Surfactant protein D (SP-D) is a member of the family of mammalian C-type lectins. SP-D is secreted into the pulmonary airspaces by lung epithelial cells and is believed to contribute to the lung's defense against inhaled microorganisms. We have previously characterized cDNAs specific for human SP-D (hSP-D). We now describe the partial characterization of genomic clones for hSP-D and present evidence for an SP-D gene with coding sequences spanning > 11 kilobases on the long arm of chromosome 10. Genomic sequencing demonstrated that the signal peptide/amino-terminal domain, the carbohydrate recognition domain, and the linking sequence between the collagen domain, and carbohydrate recognition domain are each encoded by a single exon, as for surfactant protein A and the mannose-binding protein C. However, sequencing also demonstrated a unique intron-exon structure for the collagen domain which is encoded on five exons, including four tandem exons of 117 bp. The latter exons show marked conservation in the predicted distribution of hydrophilic amino acids, consistent with tandem replication of this collagen gene sequence during evolution. Segregation analysis of HindIII digests of genomic DNA using specific cDNA probes demonstrated selective hybridization of radiolabeled hSP-D cDNA to chromosome 10- and 10q containing human/hamster somatic hybrids. The presence of SP-D gene sequences was confirmed by DNA amplification using oligomers specific for sequences within the collagen domain of the hSP-D gene. Fluorescence in situ hybridization of metaphase chromosomes using genomic probes gave selective labeling of 10q22.2 23.1. We speculate that SP-D is encoded at a locus on 10q that includes the genes for surfactant protein A. PMID- 8428972 TI - Expression of biologically active recombinant keratinocyte growth factor. Structure/function analysis of amino-terminal truncation mutants. AB - Keratinocyte growth factor (KGF) is a newly identified member of the fibroblast growth factor (FGF) family (FGF-7). KGF is expressed by stromal fibroblasts and acts on epithelial cells in a paracrine mode. To facilitate structure/function studies, we utilized the T7 prokaryotic expression system to synthesize this growth factor. Recombinant KGF (rKGF) was mitogenic with a specific activity around 10-fold higher than native KGF. By in vitro mutagenesis, we generated a series of KGF mutants with sequential deletions of the amino-terminal domain, the most divergent region among different FGF members. Mutant proteins, produced in bacteria, were tested for their ability to bind heparin, bind and activate the KGF receptor, and induce DNA synthesis. Heparin binding properties were preserved with deletion of up to 28 amino-terminal residues of the mature KGF but lost by the deletion of an additional 10 residues. Biological activity of mutants with deletions of up to 10 residues was comparable to that of rKGF. However, deletion of 29 residues resulted in significantly reduced ability to stimulate KGF receptor tyrosine-kinase activity and DNA synthesis, although this mutant bound the receptor at high affinity. These characteristics of a partial agonist may be useful in the development of competitive antagonists of KGF action. PMID- 8428973 TI - Expression of native and truncated forms of the human integrin alpha 1 subunit. AB - We report here the molecular cloning of cDNAs encoding for the human integrin alpha 1 subunit. The sequence is characteristic of an I domain containing integrin alpha subunit, with a high degree of homology to the rat integrin alpha 1 subunit, including complete identity of the transmembrane and cytoplasmic domains between the two species. The human cDNA directs the expression in mouse NIH 3T3 cells of authentic human alpha 1 protein as demonstrated by the reactivity of this subunit with two human-specific anti-alpha 1 monoclonal antibodies. This exogenous integrin specifically binds to type IV collagen in a Mg(2+)-dependent fashion. We have expressed in both transient systems and in stable cell lines truncated, soluble forms of the human alpha 1 subunit combined with truncated, soluble forms of beta 1 subunits. Although soluble beta 1 subunit was found in the media when the corresponding cDNA was used, the secretion of the soluble alpha 1 subunit was found to be dependent on dimerization with soluble beta 1. Co-transfection of truncated human alpha 1 cDNA with truncated forms of either the human or avian beta 1 cDNA led to efficient secretion of alpha 1.beta 1 heterodimers. These soluble heterodimers specifically bind to collagen IV in a manner similar to their full-length counterparts. Biosynthetic studies using stably expressing cell lines demonstrate that the soluble heterodimers and the native heterodimers are formed independently, strongly suggesting that the transmembrane or cytoplasmic domains of alpha and beta subunits are involved in the assembly of native heterodimers. PMID- 8428974 TI - Inhibition by barbiturates of the binding of Bm3R1 repressor to its operator site on the barbiturate-inducible cytochrome P450BM-3 gene of Bacillus megaterium. AB - In our previous publication (Shaw, G.-C., and Fulco, A. J. (1992) J. Biol. Chem. 267, 5515-5526), we reported that Bm3R1, a protein encoded in an open reading frame just upstream from the cytochrome P450BM-3 gene, is a repressor critically involved in the barbiturate-inducible expression of this gene in Bacillus megaterium. We now describe the purification of the Bm3R1 protein from an overproducing Escherichia coli strain harboring a bm3R1 gene-carrying plasmid and report the effect of barbiturate inducers on the interaction of Bm3R1 with a fragment of B. megaterium DNA containing the bicistronic operator and promoter sequences. Gel filtration analysis revealed that, under our experimental conditions, most of the Bm3R1 protein exists in highly aggregated forms. Gel mobility shift assays showed that Bm3R1 protein bound specifically to a segment of DNA containing the promoter-operator region of the bm3R1 gene while DNase I footprinting experiments established that Bm3R1 protected a region of DNA that covers and flanks the palindromic operator sequence. The interaction between Bm3R1 repressor and its operator, in vitro, was strongly inhibited by the addition of 2 mM pentobarbital or 2 mM methohexital (strong in vivo inducers of P450BM-3) but not by the same concentration of phenobarbital (a relatively weak inducer) or by mephobarbital (a non-inducer). A detailed comparison of pentobarbital and methohexital at concentrations lower than 2 mM indicated that methohexital was 5-10 times more effective as an inhibitor of Bm3R1 binding in vitro, as compared with its 7-fold greater inducer potency in vivo. The observation that the in vitro inhibition effects of barbiturates on the interaction of Bm3R1 repressor and its operator correlate strongly with their in vivo potency as inducers of cytochrome P450BM-3 suggests a mechanism for the induction process. It seems plausible that the barbiturate inducers might bear a conformational resemblance to and mimic the mode of action of an as yet unidentified endogenous inducer(s) in B. megaterium that functions by releasing the binding of Bm3R1 repressor from its operator site. PMID- 8428975 TI - Regulation of eukaryotic protein synthesis by initiation factors. PMID- 8428976 TI - beta-Amyloid peptide and a 3-kDa fragment are derived by distinct cellular mechanisms. AB - We have analyzed the cellular processing pathways which produce the 4-kDa amyloid beta-peptide (A beta) and a 3-kDa derivative (p3) of the beta-amyloid precursor protein (beta APP) found in conditioned media of tissue culture cells and in cerebrospinal fluid. Pulse-chase experiments reveal that both peptides are secreted in parallel with soluble beta APP (APPs); no precursor-product relation between A beta and p3 was found. The protease inhibitor leupeptin did not influence the production of either peptide. In contrast, the weak base ammonium chloride (NH4Cl) showed a dose-dependent inhibition of A beta production with less decrease in p3. A similar effect was observed using the monovalent ionophore monensin. Brefeldin A completely inhibited the generation of both peptides, indicating that proteases located in the endoplasmic reticulum or early Golgi are not sufficient for the production of the small peptides. Deletion of the beta APP cytoplasmic domain, which removes a consensus sequence that probably mediates reinternalization, caused an increase in secretion of both APPs and p3 and did not abolish A beta production. These observations suggest that completely mature beta APP within the late Golgi and/or at the cell surface is a prerequisite for A beta production but processing within the lysosome might not be directly required. p3 appears to derive from the 10-kDa C-terminal stub of beta APP following secretion of APPs. PMID- 8428977 TI - Mechanisms of collagen trimer formation. Construction and expression of a recombinant minigene in HeLa cells reveals a direct effect of prolyl hydroxylation on chain assembly of type XII collagen. AB - Collagen types IX, XII, and XIV are characterized by the presence of a highly conserved region comprising the most C-terminal triple helical domain (COL1, approximately 100 residues/chain) and 2 cysteines separated by 4 amino acid residues at the junction between this COL1 domain and the C-terminal non-triple helical domain (NC1). In order to better understand the functions of this conserved domain, we have constructed a recombinant minigene, comprising the sequence coding for an unrelated signal peptide and for the COL1 and NC1 domains of type XII collagen. This construct was placed under the control of the cytomegalovirus promoter and transfected into HeLa cells. The cells expressed the transfected minigene and the secreted chain, called alpha 1 (mini XII), could be detected by immunotransfer with an anti-peptide antibody recognizing an epitope found in the NC1 domain. Under conditions preventing the hydroxylation of prolyl residues (absence of ascorbate or presence of alpha alpha'-dipyridyl), interchain disulfide bridges did not form, while in the presence of ascorbate, disulfide bonded (alpha 1 (mini XII))3 molecules were secreted. The collagenous nature and triple helical conformation of the trimeric molecule were ascertained by the differential resistances of the COL1 and NC1 domains to trypsin and collagenase digestions, respectively. Our data demonstrate that the NC1 and COL1 domains of type XII collagen contain the information necessary for trimer formation and that, contrary to the fibrillar collagen types, posttranslational modification of the triple helical domain is essential for assembly and disulfide bonding of the chains. PMID- 8428978 TI - High affinity binding of human vitamin K-dependent protein S to a truncated recombinant beta-chain of C4b-binding protein expressed in Escherichia coli. AB - C4b-binding protein (C4BP) is a plasma glycoprotein that contains independent binding sites for complement component C4b and anticoagulant vitamin K-dependent protein S. C4BP is composed of seven alpha-chains (70 kDa) and one beta-chain (45 kDa) joined by disulfide bonds. In addition to non-repeat regions, the alpha- and beta-chains contain eight and three tandemly arranged modules, respectively, designated short consensus repeats (SCRs). C4b binds to the alpha-chains while it has been suggested, though not conclusively shown, that the beta-chain binds protein S. A truncated recombinant beta-chain composed of the three SCRs was expressed as a fusion protein with an epitope for a calcium-dependent monoclonal antibody (HPC4) in a procaryotic expression system. A signal peptide from the pelB gene of Erwinia carotovora directed expression to the medium from which the recombinant protein was purified using a HPC4 affinity column. On unreduced polyacrylamide gel electrophoresis in the presence of SDS, the recombinant protein demonstrated two closely spaced bands (M(r) = 19,000 and 20,000), whereas after reduction a broad band with an apparent molecular weight of 25,000 was obtained. On 125I-protein S ligand blotting, only one of the unreduced bands was found to bind protein S. The protein corresponding to this band bound protein S with almost as high affinity as intact C4BP, and it completely inhibited binding of protein S to intact C4BP. After reduction, the protein S binding ability was lost. The multiple carbohydrate side chains present in the native beta-chain of C4BP were apparently not required for protein S binding, as the recombinant beta chain was not glycosylated. These results demonstrate that the protein S binding site of C4BP is contained within the three SCR modules of its beta-chain. PMID- 8428979 TI - Dexamethasone-induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation. AB - Apoptosis is a major form of cell death, characterized morphologically by chromatin condensation and biochemically by endonuclease cleavage of DNA into oligonucleosomal fragments. Recently, we reported that zinc arrested dexamethasone-induced apoptosis in thymocytes at an early stage, as characterized morphologically by condensation of heterochromatin into clumps abutting the nuclear membrane. In this study, we show that zinc completely inhibits endonuclease cleavage of DNA into oligonucleosomal fragments but does not prevent the cleavage of DNA into high molecular weight fragments. These results indicate that the formation of these high molecular weight fragments, which correlates with the very early morphological features of apoptosis, is a critical event in glucocorticoid-induced apoptosis. The formation of these high molecular weight fragments, despite the inhibition by zinc of the endonuclease cleavage of DNA, suggests that key enzyme(s), other than the Ca2+/Mg(2+)-dependent endonuclease, are involved at the earliest stages of induction of apoptosis. PMID- 8428980 TI - Brefeldin A causes a defect in secretion in Saccharomyces cerevisiae. AB - Brefeldin A (BFA) blocks secretion in mammalian cells and causes the redistribution of Golgi resident membrane proteins to the endoplasmic reticulum (Klausner, R. D., Donaldson, J. G., and Lippincott-Schwartz, J. (1992) J. Cell Biol. 116, 1071-1080). The target(s) of BFA and its mechanism of action remain unknown. The yeast Saccharomyces cerevisiae represents an ideal organism in which to identify the BFA targets, since many molecules essential for vesicular traffic have been already identified taking advantage of the powerful genetics of this system. Unfortunately, wild type S. cerevisiae strains are largely insensitive to BFA (Hayashi, T., Takatsuki, A., and Tamura, G. (1982) Agric. Biol. Chem. 46, 2241-2248). Here we demonstrate that an erg6 mutant (Gaber, R., Copple, D., Kennedy, B., Vidal, M., and Bard, M. (1989) Mol. Cell. Biol. 9, 3447-3456) defective in the biosynthesis of ergosterol is sensitive to BFA. Treatment of erg6 cells with BFA results in an arrest in growth and causes a block in secretion similar to that seen in mammalian cells treated with BFA. Our data suggest that the changes in the erg6 strain allows BFA entry and that this strain can be used to examine the molecular mechanism of BFA action. PMID- 8428981 TI - Interactions of a hybrid insulin/insulin-like growth factor-I analog with chimeric insulin/type I insulin-like growth factor receptors. AB - We have examined, by use of a hybrid insulin/insulin-like growth factor-I analog and chimeric insulin/type I insulin-like growth factor receptors, the interplay between ligand and receptor structure in determining the affinity and specificity of hormone-receptor interactions in the insulin and insulin-like growth factor-I systems. Our findings, obtained through the study of radiolabeled peptide binding to detergent-solubilized full-length receptors and to soluble truncated receptors, show that (a) the two-chain hybrid analog exhibits significant cross reactivity with both receptor systems, (b) the exchange of appropriate domains in chimeric receptors enhances the receptor binding affinity of the analog by 3.5-21 fold, and (c) the affinity of the hybrid analog for the chimeric receptors actually exceeds that of either natural insulin or natural insulin-like growth factor-I. We conclude that the specificity-conferring domains of the insulin and type I insulin-like growth factor receptors reside in different regions of a common binding site, and that the exchange of domains between pairs of related hormones and between pairs of related receptors can yield new ligand-receptor systems with significantly altered affinities and selectivities. PMID- 8428982 TI - Carbohydrate residues modulate the activation of coagulation factor X. AB - Factor X is a plasma protein involved in both the intrinsic and extrinsic pathways of blood coagulation. Post-translational modifications of the protein involve gamma-carboxylation of specific glutamic acid residues, beta hydroxylation of one aspartic acid residue, and N- and O-linked glycosylation. Even though it is known that gamma-carboxylation is instrumental in regulating biological activity, the role of glycosylation in the function and properties of factor X has not been previously investigated. We utilized lectin binding and glycosidase treatment to investigate the functional role of carbohydrates on the activation peptide of factor X. Sambucus nigra agglutinin, a lectin that binds to sialic acid terminally linked alpha(2-6) to galactose or N-acetyl-galactosamine inhibits activation of human factor X in a dose-dependent manner. Inhibition of activation was observed for both intrinsic (factor IXa/VIIIa) and extrinsic (factor VIIa/tissue factor) pathway complexes. In accordance with this, selective removal of sialic acid residues on the activation peptide of factor X by neuraminidase also results in a drastic reduction of activation of the zymogen by these complexes. Corresponding reduction of activity in classical clotting assays (activated partial thromboplastin time and prothrombin time) also agrees with this observation. These results suggest a possible role of N-linked carbohydrates in the activation of factor X. PMID- 8428983 TI - Cloning and sequence analysis of cDNAs encoding precursors of sarafotoxins. Evidence for an unusual "rosary-type" organization. AB - Sarafotoxins (SRTXs) are 21-amino acid peptides structurally and functionally similar to endothelins (ETs). To understand how SRTXs are overproduced in venom glands of the snakes Atractaspis engaddensis and hence used as toxins, we cloned cDNAs encoding SRTXs and elucidated their nucleotide sequences. We predict that SRTX precursors are large prepropolypeptide chains with an unusual "rosary-type" structure made of 12 successive similar stretches of 40 residues (39 in the first stretch). Each stretch begins with a "spacer" of 19 invariant residues (18 in the first stretch) immediately followed by the sequence of one SRTX isoform. Six different isoforms are identified within a single precursor molecule. Maturation of the precursor may require endopeptidases that cleave the Leu-Cys bond and the Trp-Arg/Lys bond invariably found at the SRTX N and C termini, respectively. PMID- 8428984 TI - Physical and kinetic characterization of the DNA packaging enzyme from bacteriophage lambda. AB - Terminases are enzymes common to complex double-stranded DNA viruses and are required for packaging of the viral genome into a preformed capsid. The overexpression of bacteriophage lambda-terminase in Escherichia coli has been previously reported (Chow, S., Daub, E., and Murialdo, H. (1987) Gene (Amst.) 60, 277-289), and we present here a purification scheme for the isolation of milligram quantities of protein which is homogenous ( > 97%) as determined by SDS polyacrylamide gel electrophoresis. lambda-Terminase is composed of the gene products of Nu1 and A. Using N-terminal amino acid sequence analysis of the purified protein, we have determined a subunit stoichiometry of 2 gpNu1 polypeptides/gpA molecule in terminase holoenzyme. The circular dichroism spectrum for the purified holoenzyme has been obtained and is consistent with a protein complex composed primarily of alpha-helical structure. The endonucleolytic activity of the enzyme (the TER reaction) has been optimized with respect to pH, salt, and polyamine concentrations. Divalent metal ion is strictly required for the reaction and may be satisfied by either magnesium or manganese, but not by any of the other metals examined. E. coli integration host factor in amounts stoichiometric with the DNA substrate stimulates the TER reaction, but only when the enzyme is present in limiting amounts. Increasing the enzyme/DNA ratio attenuates the observed stimulation by integration host factor. A kinetic analysis of the TER reaction suggests that the assembly of multiple terminase promoters is required for efficient cleavage of viral DNA and that this reaction appears to be stoichiometric, rather than catalytic under the reaction conditions utilized. The implications of these results with respect to the packaging of viral DNA by terminase enzymes are discussed. PMID- 8428985 TI - A 5' control region of the human epsilon-globin gene is sufficient for embryonic specificity in transgenic mice. AB - When introduced as part of DNA constructions containing the human beta-globin locus control region (LCR), the human embryonic beta-globin gene, epsilon, is expressed in primitive but not definitive erythroid cells of recipient transgenic mice. In contrast to this pattern, the human fetal beta-globin gene, gamma, has been shown to be expressed in both primitive and definitive erythroid cells of transgenic mice when introduced in similar LCR-containing constructions. To begin to identify the minimal sequence(s) necessary for the epsilon expression pattern, we have fused a DNA fragment that contains the human epsilon-globin gene promoter region, and 13.7-kilobase (kb) of contiguous upstream flanking sequence containing super-hypersensitive (HS) sites 5'HS-2 and 5'HS-1 of the globin LCR, to the structural portion and near 3'-flanking region of the human gamma-globin gene. This construction, and one containing an intact human gamma-globin gene with the same 3'-flanking sequence and 383 base pairs of 5'-flanking sequence linked to LCR DNA from -0.86 to -13.7 kb upstream of epsilon, were each microinjected to produce transgenic mice. While the construction containing the intact gamma-globin gene is transcriptionally active in primitive and definitive erythroid cells of the transgenic mice, the fusion construction, in which the gamma-globin gene promoter and promoter proximal region is essentially replaced by that of epsilon, is not active in definitive erythroid cells and expresses with the same pattern as an intact epsilon gene. These results indicate that the promoter and near 5'-flanking region of epsilon, when linked to the LCR, is sufficient for embryonic-specific expression in transgenic mice. The level of expression of the fusion construction in primitive erythroid cells of transgenic mice is similar to that previously observed for the intact epsilon gene when identically cloned. This suggests that the epsilon 5'-region of the fusion construction also contains all the sequence necessary for the LCR-dependent activation of epsilon in transgenic mice. PMID- 8428986 TI - Structural alterations in the peptide backbone of beta-amyloid core protein may account for its deposition and stability in Alzheimer's disease. AB - The structure of beta-amyloid (beta A) from Alzheimer disease brains was examined to determine if post-translational modifications might be linked to the abnormal deposition of this peptide in the diseased tissue. The beta A peptides were isolated from the compact amyloid cores of neuritic plaques and separated from minor glycoprotein components by size-exclusion high-pressure liquid chromatography (HPLC). This parenchymal beta A has a maximal length of 42 residues, but shorter forms with "ragged" NH2 termini are also present. Tryptic peptide analysis revealed heterogeneity in the beta A1-5 and beta A6-16 peptides, each of which eluted as four peaks on reverse phase HPLC. Amino acid composition and sequence analyses, mass spectrometry, enzymatic methylation, and stereoisomer determinations revealed that these multiple peptide forms resulted from structural rearrangements of the aspartyl residues at beta A positions 1 and 7. The L-isoaspartyl form predominates at each of these positions, whereas the D isoaspartyl, L-aspartyl, and D-aspartyl forms are present in lesser amounts. beta A purified from the leptomeningeal microvasculature contains the same structural alterations as parenchymal beta A, but is 2 residues shorter at its COOH terminus. Using two different purification protocols, and using a synthetic beta A1-42 peptide as a control, we show that these modifications arose endogenously and were not caused by the experimental manipulations. The abundance of structurally altered aspartyl residues may profoundly affect the conformation of the beta A protein within plaque cores and thus significantly impact normal catabolic processes designed to limit its deposition. These alterations may therefore contribute to the production and stability of beta-amyloid deposits in Alzheimer brain tissue. PMID- 8428987 TI - Identification of the mitochondrial branched chain aminotransferase as a branched chain alpha-keto acid transport protein. AB - Conditions were developed which optimized reconstitution of branched chain alpha keto acid transport activity, which was measured as alpha-ketoisocaproate (KIC) transport, and pyruvate transport activity. Reconstitutable KIC transport activity was about 40-fold higher in heart than in liver mitochondrial extracts and 40-fold higher than heart pyruvate transport activity but only about 7-fold higher than liver pyruvate transport activity. A purification procedure was developed for the branched chain alpha-keto acid and pyruvate transport proteins which resulted in partial purification of the proteins from rat heart mitochondria. Pyruvate transport activity appeared to be associated with a 32.5 kDa protein, whereas KIC transport activity appeared to be associated with two proteins around 41 kDa. As shown by immunoblotting and immunoaffinity chromatography, these proteins were recognized by an antiserum raised against purified rat heart mitochondrial branched chain aminotransferase (BCATm). Procedures used to extract BCATm from mitochondria effectively solubilized KIC transport activity, and both transaminase and transport activities in the sonicate supernatant were immunoprecipitated by BCATm antiserum. The tissue distribution of reconstitutable KIC transport activity was identical with the tissue distribution of BCATm. On the other hand, the distribution of reconstitutable pyruvate transport activity was distinct from that of KIC transport and branched chain aminotransferase activities, and pyruvate transport activity could not be immunoprecipitated by BCATm antiserum. When incorporated into phospholipid vesicles, purified BCATm exhibited branched chain alpha-keto acid transport activity but did not transport pyruvate. Transport was saturable, and Km values for KIC and alpha-ketoisovalerate uptake were 10 and 25 microM, respectively. Substrate competition experiments indicated that KIC transport could be inhibited substantially by branched chain alpha-keto acids and their derivatives but not by substrates for the pyruvate transporter. Transport was also inhibited by several aromatic carboxylic acid derivatives and alpha ketoglutarate, but not by N-butylmalonate and succinate. Studies with covalent protein-modifying reagents indicated that transport was inhibited by sulfhydryl reagents, the histidine reagent diethyl pyrocarbonate, and the tyrosine reagent N acetylimidazole. When BCATm was incorporated into phospholipid vesicles, pyridoxal 5-phosphate was an inhibitor of transport (75% inhibition at 10 mM) but had little effect on aminotransferase activity. The data indicate BCATm is a bifunctional protein catalyzing branched chain amino acid transamination and branched chain alpha-keto acid transport. The transport properties of BCATm suggest that this protein may be the branched chain alpha-keto acid transporter that was originally identified and characterized kinetically in isolated rat heart mitochondria (Hutson, S.M., and Rannels, S.L. (1985) J. Biol. Chem. 260, 14189-14193). PMID- 8428988 TI - Glutamate synthase genes of the diazotroph Azospirillum brasilense. Cloning, sequencing, and analysis of functional domains. AB - A 10-kilobase EcoRI fragment of Azospirillum brasilense genomic DNA was cloned in Escherichia coli. Two open reading frames of 4548 and 1446 base pairs (bp) were identified within the fragment as the structural genes for the alpha and beta subunits (gltB and gltD, respectively) of A. brasilense GltS. The organization of the gltBD region of A. brasilense differs from that of the corresponding region in E. coli: in A. brasilense, gltD is upstream relative to gltB, and its stop codon is separated by 141 bp from the first ATG of gltB. The deduced amino acid sequences reveal a high similarity with GltS from E. coli and with the ferredoxin dependent GltS from maize. Binding domains for flavin cofactors and NADPH, a domain for glutamine binding and activation, and cysteine clusters for iron sulfur centers formation were tentatively identified on the basis of sequence comparison with flavoproteins, pyridine nucleotide-dependent enzymes, amidotransferases, and iron-sulfur proteins. PMID- 8428989 TI - Inactivation of the recA protein by mutation of histidine 97 or lysine 248 at the subunit interface. AB - We have used site-directed mutagenesis to prepare two new mutant recA proteins, one in which histidine 97 has been replaced by alanine, and another in which lysine 248 has been replaced by alanine. Although these mutant proteins were originally designed from different considerations, they turned out to have remarkably similar properties. Both the [H97A]recA protein and the [K248A]recA protein bind poorly to single-stranded DNA, have no single-stranded DNA-dependent ATP hydrolysis activity, and do not promote renaturation of complementary single stranded DNA molecules or the ATP-dependent three-strand exchange reaction. Furthermore, both mutant proteins are defective in Mg(2+)-induced helical filament formation. To account for these results, we propose that the mutation of either histidine 97 or lysine 248 alters subunit interactions between recA monomers and that this leads to the loss of cooperative single-stranded DNA binding and DNA pairing activities. This proposal is consistent with the recently determined x-ray structure of the recA protein, which shows that although histidine 97 and lysine 248 are distant from one another in the monomer structure, these two residues are on the opposing complementary faces of the recA subunit and pack against each other at the interface between adjacent recA monomers in the helical filament (Story, R. M., Weber, I. T., and Steitz, T. A. (1992) Nature 355, 318-325). PMID- 8428990 TI - High density lipoprotein interconversions in rat and man as assessed with a novel nontransferable apolipopeptide. AB - A nontransferable peptide analog of a plasma apolipoprotein diacyl lipid associating peptide (diLAP) incorporates into model reassembled high density lipoproteins (R-HDL). In whole plasma in vitro, diLAP irreversibly transfers to native rat HDL2 and human HDL3, but not to rat HDL1 or human HDL2. The rate of transfer is dependent on the physical state of the lipid in the R-HDL. Exogenous cholesterol promotes the formation of larger HDL. When diLAP-labeled R-HDL were injected into rats, the diLAP that initially associated with HDL2 transferred to HDL1 over a period of 48 h. The rate of clearance of diLAP-labeled HDL was slower than that of apoA-I. The liver was the preferred site for diLAP-labeled HDL1 uptake. In contrast, diLAP-labeled HDL2 were associated with liver, ovaries, and adrenal glands, with the adrenal grands exhibiting the highest specific association. DiLAP was not found in the kidneys. These data show that 1) rat HDL is cleared more slowly than rat apoA-I; 2) HDL is removed from the plasma compartment as a particle; 3) there are tissue-specific differences in the removal of rat HDL1 and HDL2; 4) HDL2 is a precursor to HDL1; and 5) cholesterol and the activity of lecithin:cholesterol acyltransferase are essential to HDL remodeling. PMID- 8428991 TI - Detergent insolubility of alkaline phosphatase during biosynthetic transport and endocytosis. Role of cholesterol. AB - Alkaline phosphatase is anchored to the outer leaflet of the plasma membrane by a covalently attached glycosyl-phosphatidylinositol anchor. We have studied the biosynthetic transport and endocytosis of alkaline phosphatase in the choriocarcinoma cell line BeWo, which endogenously expresses this protein. It was demonstrated that the protein was synthesized as a Triton X-100-soluble precursor. During transport to the cell surface the enzyme was converted in a mature form, which was insoluble in Triton X-100 at 0 degrees C. Once at the cell surface 85% of alkaline phosphatase remained in the detergent-insoluble form. Under steady state conditions 15% of alkaline phosphatase was endocytosed. Most interestingly, this fraction of internalized alkaline phosphatase was completely soluble in Triton X-100 at 0 degrees C. After depletion of membrane cholesterol by saponin, alkaline phosphatase became completely soluble in Triton X-100 at 0 degrees C, suggesting that cholesterol plays a critical role in the formation and maintenance of Triton X-100-resistant membrane domains. PMID- 8428992 TI - Domains near ATP gamma phosphate in the catalytic site of H+-ATPase. Model proposed from mutagenesis and inhibitor studies. AB - The beta Gly-149 residue is in a glycine-rich sequence (Gly-Gly-Ala-Gly-Val-Gly Lys-Thr; residues 149-156) of the Escherichia coli H(+)-ATPase (ATP synthase) beta subunit. Substitution of beta Gly-149 by Ser suppressed the effect of the beta Ser-174-->Phe mutation (Iwamoto, A., Omote, H., Hanada, H., Tomioka, N., Itai, A., Maeda, M., and Futai, M. (1991) J. Biol. Chem. 266, 16350-16355), suggesting that beta Gly-149 is located near beta Ser-174. In this study, we introduced different residues at position 149 and found that a single mutant beta Cys-149 was defective. The effect of beta Cys-149 mutation was suppressed by beta Gly-172-->Glu, beta Ser-174-->Phe, beta Glu-192-->Val, or beta Val-198-->Ala replacement. These results suggest that beta Gly-149, beta Gly-172, beta Ser-174, beta Glu-192, and beta Val-198 residues are located close together in the catalytic site. From these findings we propose a model of the catalytic site of the enzyme near the gamma phosphate moiety of ATP. F1 enzymes with the double mutations beta Cys-149/beta Glu-172, beta Cys-149/beta Phe-174, beta Cys-149/beta Val-192, and beta Cys-149/beta Ala-198 were less sensitive than wild-type F1 to dicyclohexylcarbodiimide and adenosine triphosphopyridoxal (an affinity analogue of ATP forming a Schiff base with the epsilon-amino group of beta Lys-155 or beta Lys-201), and became sensitive to N-ethylmaleimide in an ATP-protected manner. These results of inhibitor studies are consistent with the proposed model. PMID- 8428993 TI - Purification and analysis of streptococcal NADH peroxidase expressed in Escherichia coli. AB - Using the T7 RNA polymerase expression system, a modified plasmid vector has been developed which gives reliable, high level expression in Escherichia coli of the gene encoding streptococcal NADH peroxidase. The recombinant enzyme has been purified to homogeneity using a revised protocol which yields over 35 mg of pure protein per liter of culture. Recombinant NADH peroxidase is fully active and exhibits spectroscopic and redox properties identical to those for the enzyme purified from Streptococcus faecalis 10C1. Reductive titrations and thiol analyses confirm the presence of the unusual cysteine-sulfenic acid (Cys-SOH) redox center identified previously. N-terminal sequence analysis, analytical gel filtration, and preliminary x-ray diffraction data all confirm the structural identity of the recombinant and S. faecalis enzymes. Steady-state kinetic analysis of the peroxidase, coupled with results from static titration experiments is consistent with a limiting type of ternary complex mechanism and allows the determination of many of the corresponding kinetic constants. In addition, preliminary 1H NMR spectra of the enzyme at millimolar concentrations show good dispersion in the amide region and indicate that the recombinant peroxidase is suitable for one-dimensional NMR work with labeled amino acids. PMID- 8428994 TI - Lipooligosaccharidic antigen containing a novel C4-branched 3,6-dideoxy-alpha hexopyranose typifies Mycobacterium gastri. AB - Lipooligosaccharides (LOSs) have recently been proposed as markers of mycobacterial avirulence. They have been characterized in Mycobacterium kansasii cell wall and were investigated in Mycobacterium gastri since the distinction between the two mycobacterial species remains in some question. A set of unknown LOSs was isolated from M. gastri W471. The highly antigenic lipooligosaccharide, LOS-III, was purified and appeared in all the M. gastri strains investigated regardless of their morphology. Moreover, by enzyme-linked immunosorbent assay and chromatographic approaches, it was found that LOS-III unambiguously distinguished M. gastri from the opportunistic pathogen M. kansasii. The LOS-III structure was established from its native form using NMR spectroscopy. This strategy revealed the presence of a supplementary monosaccharide (X) which was not characterized by routine carbohydrate analysis. Its core structure, 3,6 dideoxy-alpha-hexopyranose, was established from the complete assignment of the 1H and 13C spectra by two-dimensional homonuclear (COSY, HOHAHA) and heteronuclear 1H-13C heteronuclear multiple quantum correlation spectroscopy (HMQC) and HMQC-HOHAHA spectroscopy. Due to the absence of a proton at C4, the key data of the C4 side chain structure came from the heteronuclear multiple bond correlation spectroscopy (HMBC) spectrum. It was revealed to be a C-alkyl chain of partial structure 1,3-dimethoxypropyl. From the HMBC spectrum, this novel C branched monosaccharide was located at the nonreducing end of the LOS, while the putative reducing end was found to consist of a 2',4,6-triacylated alpha-alpha trehalose. The following structure, based on the evidence presented in this paper, is proposed for LOS-III: Xp alpha(1-->3)[L-Xylp beta(1-->4)](6)3-O-Me-Rhap alpha(1-->3)D- Galp beta(1-->3)-D-Glcp beta(1-->4)2-O-acyl-D-Glcp alpha(1<==>1)alpha 4,6-di-O- acyl-D-Glcp. PMID- 8428995 TI - Lymphoid V(D)J recombination. Functional analysis of the spacer sequence within the recombination signal. AB - The V(D)J recombination reaction is directed by a pair of signal sequences, each consisting of a palindromic heptamer, an A/T-rich nonamer, and an intervening spacer sequence of 12 or 23 base pairs. The spacer sequence previously has not been analyzed for a functional role. In this study, numerous simultaneous sequence changes have been made in the spacer of each signal to test their functional importance. All of the AT base pairs in each signal were changed to GC base pairs. This particular change is of interest because it markedly increases the energy that would be required to melt out the two strands of each signal to permit the intersignal base pairing proposed in a commonly invoked model for signal-signal interaction in V(D)J recombination. We find that changing 6 to 12 AT base pairs in the 12-signal to GC does not affect V(D)J recombination, nor does changing 11 of 23 AT base pairs in the 23-signal. Substrates with all-GC spacer sequences in both the 12- and the 23-signal also recombine at efficiencies that are not significantly reduced. These studies demonstrate that the sequences at these particular positions are not recognized by the recombinase. In addition, the data do not support models invoking signal-signal base pairing. PMID- 8428996 TI - Preferential degradation of protein-bound (ADP-ribose)n by nuclear poly(ADP ribose) glycohydrolase from human placenta. AB - Poly(ADP-ribose) glycohydrolase, extensively purified to homogeneity from nuclei of human placenta, is composed of a single polypeptide with a molecular mass of 71,000 daltons on sodium dodecyl sulfate-polyacrylamide gel. Judging from its physico-chemical and catalytic properties, the enzyme is similar to the nuclear glycohydrolase (glycohydrolase I), but not to the cytoplasmic glycohydrolase (glycohydrolase II) that has been purified from guinea pig liver (Tanuma, S., Kawashima, K., and Endo, H. (1986) J. Biol. Chem. 261, 965-969; Maruta, H., Inageda, K., Aoki, T., Nishina, H., and Tanuma, S. (1991) Biochemistry 30, 5907 5912). The rates of hydrolysis of (ADP-ribose)n bound to various proteins by the purified nuclear glycohydrolase were higher than those of the corresponding free polymers. Kinetic analyses revealed that the enzyme had more activity toward poly(ADP-ribose) bound to histone H1 or to poly(ADP-ribose) polymerase than toward oligo(ADP-ribose) bound to cytoplasmic proteins from mitochondria or mRNA ribonucleoprotein although the Km and Vmax values were dependent on the chain length (n). In contrast, cytoplasmic glycohydrolase purified from human erythrocytes was more active toward oligo(ADP-ribose) (n = 2.6 or 4.2) bound to the cytoplasmic proteins than to poly(ADP-ribose) (n = 14.6) bound to histone H1, and their kinetic parameters of glycohydrolase II were rather dependent on the acceptor molecules for (ADP-ribose)n. These results suggest that poly(ADP-ribose) glycohydrolase I may play an important role in regulation of poly(ADP ribosyl)ation levels on chromosomal proteins in nuclei. PMID- 8428997 TI - Mutagenesis of acidic residues in putative membrane-spanning segments of the melibiose permease of Escherichia coli. I. Effect on Na(+)-dependent transport and binding properties. AB - Four aspartic acids, distributed in different putative membrane-spanning segments of the NH2-terminal domain of melibiose (mel) permease (D31 in helix I, D51 and D55 in helix II, and D120 in helix IV) were individually replaced by either Asn or Cys using site-directed mutagenesis. mel permease with either neutral residues at position 51, 55, or 120 or permease with a Cys in place of D31 does not catalyze significant Na(+)-linked methyl-1-thio-beta-D-galactopyranoside (TMG) accumulation. Binding studies of a high affinity ligand (p-nitrophenyl-alpha-D galactopyranoside (NPG)) on de-energized membrane vesicles indicate that these modified transporters (i) retain the ability to bind the alpha-galactosides NPG or melibiose and the beta-galactoside TMG and (ii) exhibit a Na(+)-independent sugar-binding phenotype. In contrast, mel permease with an Asn residue at position 31 mediates Na(+)-coupled TMG transport and displays a Na(+)-dependent sugar binding phenotype, but requires a higher concentration of sodium than wild type permease to produce maximal stimulation of sugar binding. The observation that individual mutation of the Asp residue at position 31, 51, 55, or 120 systematically and selectively modifies the contribution of the coupling ion to the early step of the transport reaction, i.e. cosubstrate binding, raises the possibility that (i) these 4 aspartic residues are at or near the cationic binding site of mel permease, (ii) the NH2-terminal domain of mel permease in which they are distributed accommodates or is part of the cationic binding site, and (iii) the oxygen atoms of these Asp side chains contribute to coordination of the coupling ion. PMID- 8428998 TI - Mutagenesis of acidic residues in putative membrane-spanning segments of the melibiose permease of Escherichia coli. II. Effect on cationic selectivity and coupling properties. AB - Individual substitution of Cys or Asn for Asp-31, Asp-51, Asp-55, or Asp-120, distributed in different membrane spanning segments of the NH2-terminal domain of melibiose (mel) permease partially or completely inactivates Na(+)-linked sugar transport and stimulation of sugar binding on mel permease by Na+ ions (Pourcher, T., Zani, M.-L., and Leblanc, G. (1993) J. Biol. Chem. 268, 3209-3215). To investigate further the effect of these substitutions on the cationic selectivity and coupling properties of mel permease, H(+)-melibiose coupled transport, coupling between H+ and melibiose movements, sugar counterflow, and zero-trans sugar efflux by the mutant permeases were analyzed. The results provide additional evidence indicating that manipulation of some of these Asp in the membrane-spanning segments of mel permease alters its cationic selectivity properties. The results also indicate that the individual mutations diversely affect mel permease-coupling properties. For example, only permease with Asn in place of Asp-31 or Cys in place of Asp-51 retains the capacity to actively transport melibiose. On the other hand, replacing Asp-55 by Cys produces uncoupling of cosubstrate flows by the carrier but does not hamper sugar translocation. These and other features of the mutant permeases are used to discuss the relative participation of Asp-31, Asp-51, Asp-55, or Asp-120 to the mel symport mechanism and to its ionic selectivity and also the existence of a possible gating mechanism that may contribute the obligatory coupling of cosubstrate flows by the symporter. PMID- 8428999 TI - Only one of the charged amino acids located in the transmembrane alpha-helices of the gamma-aminobutyric acid transporter (subtype A) is essential for its activity. AB - The gamma-aminobutyric acid (GABA) transporter (subtype A) is located in nerve terminals and catalyses coupled electrogenic uptake of the neurotransmitter with two or three sodium and one chloride ions. It contains 599 amino acids and 12 putative membrane spanning alpha-helices and is the first described member of a neurotransmitter transporter superfamily. The membrane domain contains 5 charged amino acids which are basically conserved. Using site-directed mutagenesis, we show that only one of them, arginine 69, is absolutely essential for activity. It is located in a highly conserved region encompassing parts of helices 1 and 2. The three other positively charged amino acids and the only negative charged one, glutamate 467, are not critical. These results suggest that the translocation pathway of the sodium ions through the membrane does not involve charged amino acid residues and underline the importance of the highly conserved stretch between amino acids 66 and 86. PMID- 8429000 TI - Specific contribution of different phospholipid surfaces to the activation of prothrombin by the fully assembled prothrombinase. AB - This paper addresses, in thermodynamic and kinetic terms, the reasons for the acidic lipid specificity of the human prothrombinase complex. We obtained, from the measured lipid titrations of the initial rates of prothrombin activation, the empirical binding constants for prothrombinase assembly on different membranes. These favored assembly on phosphatidylserine (PS)- as opposed to phosphatidylglycerol (PG)-containing membranes. In addition, we have used full time courses of prothrombin activation, in conjunction with a calculation of the equilibrium distribution of factor Xa between four enzymatic forms, to obtain the intrinsic kinetic constants of the prothrombinase assembled on PS- or PG containing membranes. The resulting values of kcat, Km, and kcat/Km increased as acidic lipid content increased, and kcat/Km reached a plateau at 12 mol % PS and 50 mol % PG. Using the measured assembly and kinetic constants, the observed shapes of the phospholipid titration curves of human prothrombin activation were interpreted. We conclude that the difference in activity of prothrombinase assembled on PS- versus PG-containing membranes results both from the different binding properties of factors Xa and Va to these surfaces and from the different intrinsic activities of the prothrombinase when assembled on different membranes. PMID- 8429001 TI - Simulation of prothrombin proteolysis by the full prothrombinase assembled on varied phospholipid surfaces. PMID- 8429002 TI - The topology of the anchor subunit of dimethyl sulfoxide reductase of Escherichia coli. AB - The terminal electron transfer enzyme Me2SO reductase of Escherichia coli is a heterotrimeric enzyme composed of a membrane extrinsic catalytic dimer (DmsAB) and a membrane intrinsic polytopic anchor subunit (DmsC). The topology of DmsC has been studied using phoA (alkaline phosphatase) and blaM (beta-lactamase) gene fusions. The results of analyzing the properties of proteins produced by the fusions suggests a structure with eight transmembrane helices. Both the amino and carboxyl termini are exposed to the periplasm. The entire DmsC polypeptide is necessary to anchor DmsAB to the membrane as fusions with truncated DmsC were not functional and soluble DmsAB accumulated in the cytoplasm. A dmsC-phoA fusion in the termination codon of dmsC generated a chimeric enzyme with functional Me2SO reductase and alkaline phosphatase activity. Quantitation of the minimal inhibitory concentration of ampicillin for the dmsC-blaM fusions indicated that different transmembrane helices had differing signal sequence activity. PMID- 8429003 TI - Site-specific glycosylation of recombinant rat and human soluble CD4 variants expressed in Chinese hamster ovary cells. AB - The rat and human forms of the T-cell surface glycoprotein CD4 share a common glycosylation site at the Asn270/271 position but differ with respect to the locations of the second glycosylation sites at Asn159 (rat) and Asn300 (human). The glycosylation of soluble recombinant forms of human and rat CD4 (sCD4) expressed in Chinese hamster ovary cells has been characterized. The most obvious differences between the rat and human sCD4 oligosaccharides were the greater abundance of oligomannose and hybrid oligosaccharides on rat sCD4 and the presence of oligosaccharides carrying a terminal alpha-galactose residue on human sCD4. This is the first report of the occurrence of alpha-galactose residues on a glycoprotein expressed in Chinese hamster ovary cells. Comparison of mutant rat sCD4 molecules with single glycosylation sites and glycopeptides indicated that site-specific and independent processing occurred at each glycosylation site. The glycosylation at the conserved site at Asn270 of rat sCD4 was identical to that seen for the equivalent site in human sCD4, and the oligomannose and hybrid structures were restricted to the nonconserved site at Asn159 in rat sCD4. However, there was more oligosaccharide processing at this site in a truncated form of rat sCD4 consisting of the two NH2-terminal domains. These results indicate that not only the local three-dimensional structure but also domain interactions can influence the processing at individual glycosylation sites. PMID- 8429004 TI - Expression and characterization of the heme-binding domain of Chlorella nitrate reductase. AB - A recombinant protein corresponding to the putative heme-binding domain of assimilatory NADH:nitrate reductase from Chlorella vulgaris has been expressed and purified from transformed Escherichia coli BL21 cells. The recombinant protein, exhibited a subunit molecular mass of approximately 10 kDa with a N terminal sequence beginning with the residues PAGA in agreement with that predicted by cDNA analysis. The UV-visible spectrum of the protein confirmed the incorporation of heme with maxima at 413 nm and 423, 528, and 557 nm for the oxidized and reduced forms, respectively. Circular dichroism spectra indicated the environment of the heme chromophore was very similar to that of the native enzyme. Potentiometric titrations of the recombinant heme domain yielded a midpoint potential of +16 mV (n = 1, pH 7), substantially higher than the values of -160 mV obtained for the native enzyme and -28 mV obtained for a previously expressed recombinant heme domain that contained part of the Mo-pterin domain. These results indicate that portions of the amino acid sequence that are involved in the formation of the Mo-pterin domain of Chlorella nitrate reductase influence the redox potential of the heme prosthetic group. PMID- 8429005 TI - Site-directed mutagenesis of a putative heparin binding domain of avian lipoprotein lipase. AB - Lipoprotein lipase (LPL) binds to heparin and heparan sulfate proteoglycans. We have employed site-directed mutagenesis to dissect one of the proposed heparin binding domains of avian LPL, which contains the sequence Arg-Lys-Asn-Arg (amino acids 281-284). Various single, double, and triple mutants of chicken LPL were constructed in order to alter the positive charge of this region. The mutant and wild-type cDNAs were subcloned into an expression vector, pRc/CMV, and expressed in Chinese hamster ovary cells. In general, the LPL mutants with a decrease in regional positive charge showed a decrease in affinity for heparin and heparan sulfate proteoglycans. The greatest effect was seen with the triple mutant, LPL 5G, in which all of the positively charged amino acids were altered to neutral residues. On a heparin-Sepharose column, LPL 5G eluted at 0.96 M NaCl compared with 1.35 M for wild-type LPL. This mutant also had the lowest specific activity with 1.5 mu eq fatty acid/micrograms/h for the cell-associated pool and with no detectable activity in the media. Wild-type cells, however, produced a lipase with a specific activity of 12.4 and 13.1 mu eq fatty acid/micrograms/h for cell associated and media lipase pools, respectively. LPL 5G also showed a decrease in affinity for the heparan sulfate proteoglycans on the cell surface of Chinese hamster ovary cells. In conclusion, the region of avian LPL between Arg281 and Arg284 does appear to be involved in heparin-binding; however, additional regions must be involved since binding was not completely abolished. In addition, specific activity of the cell-associated and secreted LPL is correlated to affinity of the enzyme for heparan sulfate chains. PMID- 8429006 TI - Heterologous expression and characterization of prolactin-like protein-A. Identification of serum binding proteins. AB - In this report, we describe the heterologous expression of prolactin-like protein A (PLP-A) in Chinese hamster ovary (CHO) cells, the characterization of recombinant PLP-A, and the identification of serum PLP-A-binding proteins. CHO cell and native placental PLP-A showed similar immunoreactive characteristics and electrophoretic mobilities. N-terminal sequencing verified the identity and purity of the recombinant PLP-A species and the site of cleavage of the signal peptide from the mature secreted PLP-A species. Recombinant PLP-A lacked activity in standardized prolactin and growth hormone in vitro bioassays. Antibodies generated to recombinant PLP-A facilitated the cellular localization of PLP-A and the identification of high molecular weight PLP-A complexes. Cross-linking analyses of radioiodinated PLP-A with serum harvested from late gestation rats indicated the presence of two major cross-linked complexes migrating under reducing conditions at 130 and 250 kDa and two minor cross-linked complexes migrating at 70 and 110 kDa. Binding of PLP-A to serum proteins was specific for PLP-A and not effectively competed by other members of the prolactin/growth hormone family. The PLP-A binding species were also found in serum from non pregnant female and male rats. PMID- 8429007 TI - QUAD, a protein from hepatocyte chromatin that binds selectively to guanine-rich quadruplex DNA. AB - The single-stranded oligomer Q, whose nucleotide sequence 5' d(TACAGGGGAGCTGGGGTAGA)-3' corresponds to the IgG switch region, forms in concentrated solutions and in the presence of alkali metal cation parallel four stranded complexes termed G4 DNA (Sen, D., and Gilbert, W. (1988) Nature 334, 364 366). We show that G4 DNA was also formed during storage of dried oligomer Q. This quadruplex complex migrated more slowly than mono-strand oligomer Q during nondenaturing gel electrophoresis, the rate of its formation depended on the mass of stored oligomer Q, and N7 positions of guanine residues were involved in its stabilization. Here we report the purification of a protein designated QUAD that binds specifically to the G4 form of oligomer Q, from non-histone protein extracts of rabbit hepatocytes. QUAD was 80-90% purified by sequential steps of column chromatography on Sepharose 6B, DEAE-cellulose, phosphocellulose, and phenyl-Sepharose. Purified QUAD migrated on SDS-polyacrylamide gel electrophoresis as a 58 +/- 2.6-kDa polypeptide and had a native molecular mass of 57 +/- 2.5 kDa as determined by Sepharose 6B gel filtration. The dissociation constant of G4 DNA binding to QUAD was in the range of 2.5 to 7.0 x 10(-9) M/liter. Excess unlabeled monostranded oligomer Q did not compete with 5'-32P labeled G4 DNA on its binding to QUAD. Further, that QUAD recognized the G4 DNA structure rather than a DNA sequence was also demonstrated by the inefficient competition on the binding of 5'-[32P]G4 DNA to QUAD by excess unlabeled single- or double-stranded DNA molecules that contained guanine clusters of different length or various other nucleotide sequences. PMID- 8429008 TI - Inhibition of dysthrombins Quick I and II by heparin cofactor II and antithrombin. AB - Heparin cofactor II and antithrombin are plasma serine proteinase inhibitors whose ability to inhibit alpha-thrombin is accelerated by glycosaminoglycans. Dysfunctional thrombin mutants Quick I (Arg67-->Cys) and Quick II (Gly226-->Val) were used to further compare heparin cofactor II and antithrombin interactions. Quick I, Quick II, and alpha-thrombin were eluted at the same salt concentration from heparin-Sepharose suggesting that the putative heparin-binding site (also termed anion binding exosite-II) is functional. Antithrombin yielded similar inhibition rates for Quick I and alpha-thrombin in the absence or presence of various amounts of heparin. Also, Quick I was inhibited similarly to alpha thrombin by heparin cofactor II in the absence of glycosaminoglycan. In contrast, glycosaminoglycan-accelerated Quick I inhibition by heparin cofactor II was greatly reduced indicating that anion binding exosite-I (where the mutation occurs in Quick I) is critical for increased inhibition by heparin cofactor II. We also found that heparin cofactor II formed a SDS-resistant bimolecular complex with Quick II and alpha-thrombin at similar rates and the rate of complex formation was accelerated in the presence of glycosaminoglycans. A three dimensional molecular model of the Quick II active site compared to alpha thrombin suggested that the heparin cofactor II Leu-Ser-reactive site sequence (P1-P1') is a compatible "pseudosubstrate" in contrast to the Arg-Ser sequence found in antithrombin. The importance of heparin cofactor II as a thrombin regulator will depend upon its ability to interact with glycosaminoglycans and the functional availability of thrombin exosites. PMID- 8429009 TI - Residues 1-254 of anthrax toxin lethal factor are sufficient to cause cellular uptake of fused polypeptides. AB - Anthrax lethal toxin is a complex of protective antigen (PA, 735 amino acids) and lethal factor (LF, 776 amino acids) that lyses certain eukaryotic cells. LF interacts with PA to gain access to the cytosol to assert its toxicity. The internalization of LF requires that PA bind to a specific membrane receptor and be cleaved by a cell-surface protease (probably furin), so as to expose a site on PA to which LF binds with high affinity. To localize LF functional domains, amino, carboxyl, and internal deletions of LF were made. Toxicity was eliminated by deletion of 40 and 47 residues from the amino and carboxyl termini, respectively. Similarly, deleting the first of the four imperfect repeats of 19 amino acids located at residues 308-383 made LF non-toxic, showing that this region is also essential for activity. To identify the minimum region of LF which is required for binding to PA, varying amino-terminal portions of LF were fused to the ADP-ribosylation domain of Pseudomonas exotoxin A. Fusion proteins containing residues 1-254 of LF were toxic when administered with PA, while those having only residues 1-198 of LF were inactive, showing that the PA-binding domain of LF lies within residues 1-254. PMID- 8429010 TI - Inactivation of glucose-6-phosphate dehydrogenase by 4-hydroxy-2-nonenal. Selective modification of an active-site lysine. AB - Incubation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides with 4-hydroxy-2-nonenal (HNE) results in a pseudo first-order loss of enzyme activity. The pH dependence of the inactivation rate exhibits an inflection around pH 10, and the enzyme is protected from inactivation by glucose 6 phosphate. Loss of enzyme activity corresponds with the formation of one carbonyl function per enzyme subunit and the appearance of a lysine-HNE adduct. The data presented in this paper are consistent with the view that the epsilon-amino group of a lysine residue in the glucose 6-phosphate-binding site reacts with the double bond (C3) of HNE, resulting in the formation of a stable secondary amine derivative and loss of enzyme activity. We have described a mechanism by which HNE may, in part, mediate free radical damage. In addition, a method for the detection of the lysine-HNE adduct is introduced. PMID- 8429011 TI - Inverse relationship between GLUT-4 phosphorylation and its intrinsic activity. AB - In this study, we examined the effect of phosphorylation on GLUT-4 function in isolated rat adipocytes. Adipocytes labeled with 32P for 2 h were incubated with parathyroid hormone (PTH) (20 ng/ml) for 60 min and then exposed to insulin (25 ng/ml) for an additional 30 min. 32P-GLUT-4 was immunoprecipitated from the plasma membrane and low density microsomal fractions, and its degree of phosphorylation was determined by autoradiography and densitometry. Results were expressed as 32P-GLUT-4 specific activity (phosphorylation/unit of protein). GLUT 4 intrinsic activity was measured using [14C]2-deoxyglucose uptake in plasma membrane vesicles. PTH significantly increased GLUT-4 phosphorylation and eliminated the insulin-stimulated dephosphorylation of GLUT-4. Western blotting revealed normal distribution of GLUT-4 before and after insulin stimulation in control and PTH-treated cells, suggesting that phosphorylation of GLUT-4 does not interfere with its recruitment to the plasma membrane. In contrast, intrinsic activity of phosphorylated GLUT-4 was significantly reduced (p < 0.01). Preincubation of adipocytes with calcium channel blocker (nitrendipine) and cyclic AMP antagonist (RpcAMP) restored GLUT-4 intrinsic activity in the PTH treated cells. In several experiments, GLUT-4 was phosphorylated in vitro in plasma membrane vesicles isolated from normal adipocytes exposed to insulin. This in vitro phosphorylation reduced GLUT-4 intrinsic activity by approximately 35% (p < 0.01). We conclude that phosphorylation of GLUT-4 significantly impairs the ability of insulin to stimulate its intrinsic activity. PMID- 8429012 TI - Cytosolic [Ca2+] homeostasis and tyrosine phosphorylation of phospholipase C gamma 2 in HL60 granulocytes. AB - Activated phagocytes produce large amounts of reactive oxygen intermediates, including peroxides. In addition to their microbicidal effect, it has recently been suggested that reactive oxygen species play a role as intracellular messengers. The mechanism of action remains unknown, but peroxides have been reported to increase tyrosine phosphorylation, an effect potentiated by vanadate. In this report we studied the effects of a combination of H2O2 and vanadate on Ca2+ homeostasis in granulocytic HL60 cells. The peroxides induced a transient elevation of cytosolic [Ca2+] associated with release from internal stores. Ca2+ mobilization was accompanied by increased generation of inositol 1,4,5 trisphosphate, implicating phospholipase C (PLC). A sizable increase in phosphotyrosine accumulation by several polypeptides in the M(r) 20,000 to 250,000 range preceded the [Ca2+] changes. We therefore considered the possibility that tyrosine phosphorylation of a phospholipase mediates the observed effects. Differentiated (granulocytic) HL60 cells did not have detectable levels of PLC gamma 1 but had substantial PLC gamma 2. Immunoprecipitation and immunoblotting experiments demonstrated that PLC gamma 2 becomes tyrosine-phosphorylated upon treatment of the cells with peroxides of vanadate. If associated with activation, such phosphorylation of PLC gamma 2 can account for the rise in [Ca2+]. Although capable of mobilizing internal Ca2+ stores, the peroxides failed to produce the sustained [Ca2+] increase predicted by the "capacitative" model. Mn2+ influx determinations indicated that this is due to impairment of divalent cation entry by the peroxides, uncoupling the plasma membrane from the internal stores. Changes in [Ca2+] homeostasis could mediate some of the messenger actions of reactive oxygen species. PMID- 8429013 TI - Analysis of inositol metabolites produced by Saccharomyces cerevisiae in response to glucose stimulation. AB - When cultures of Saccharomyces cerevisiae are grown to stationary phase in medium containing [3H] inositol, significant amounts of radioactivity can be detected in phosphatidylinositol, phosphatidylinositol 3-phosphate, phosphatidylinositol 4 phosphate, and phosphatidylinositol 4,5-bisphosphate. Addition of glucose to such cultures results in the generation of [3H]glycerophosphoinositol, [3H]glycerophosphoinositol 4-phosphate, and [3H]glycerophosphoinositol 4,5 bisphosphate in the extracellular medium. We found no evidence, however, for the stimulated formation of other inositol polyphosphates. This result suggests that glucose does not stimulate the "phospholipase C" signalling pathway established in higher eukaryotic cells but, in contrast, stimulates specific phospholipases A or B. A variety of cell division cycle (cdc) mutants have been studied to investigate the relationship between cell cycle progression and inositol metabolism in S. cerevisiae. Mutants which are defective for completion of cell cycle "START" (i.e. commitment to mitosis) show reduced formation of glycerophosphoinositol 4-phosphate and glycerophosphoinositol 4,5-bisphosphate in response to glucose. In contrast, cdc mutants which are defective in post-"START" processes show a larger glucose response than wild type cells. These results suggest that deacylation of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate may be coordinated with cell cycle control in S. cerevisiae. PMID- 8429014 TI - Calcium-induced cytotoxicity in hepatocytes after exposure to extracellular ATP is dependent on inorganic phosphate. Effects on mitochondrial calcium. AB - In isolated mitochondria extensive uptake of Ca2+ in the presence of an "inducing agent," e.g. inorganic phosphate (Pi), causes permeabilization of the mitochondrial inner membrane and a collapse of the mitochondrial membrane potential. In this study we tested whether the effect of phosphate occurs in intact hepatocytes. Rat hepatocytes were incubated with ATP to induce a sustained increase in intracellular Ca2+ ([Ca2+]i), dissipation of the mitochondrial membrane potential, and cell death (Zoeteweij, J. P., van de Water, B., de Bont, H. J. G. M., Mulder, G. J., and Nagelkerke, J. F. (1992) Biochem. J. 288, 207 213). Omission of Pi from the incubation medium delayed the loss of viability. The nonhydrolyzable ATP analog adenosine 5'-O-(thiotriphosphate) (ATP gamma S) had similar effects on [Ca2+]i and viability, but now omission of extracellular Pi completely protected against cytotoxicity. Exposure to ATP or ATP gamma S induced a large cellular uptake of Pi. With the use of video-microscopy a significant increase in mitochondrial free calcium was observed before the onset of cell death. Accumulation of mitochondrial calcium was reduced when extracellular Pi was omitted. These results suggest that, after induction of high [Ca2+]i by ATP in hepatocytes, 1) mitochondria accumulate calcium which is associated with cell toxicity and 2) intracellular Pi increases which stimulates mitochondrial calcium uptake. These observations support a calcium-dependent mitochondrial dysfunction, induced by phosphate, as a valid model for ATP-induced cytotoxicity in hepatocytes. PMID- 8429015 TI - Neutral sphingomyelinase increases the binding, internalization, and degradation of low density lipoproteins and synthesis of cholesteryl ester in cultured human fibroblasts. AB - I have investigated the effects of human urinary neutral sphingomyelinase (N SMase) (Chatterjee, S., and Ghosh, N. (1989) J. Biol. Chem. 264, 12554-12561) on the cell-surface binding, internalization, and degradation of 125I-low density lipoprotein (LDL) and on cholesteryl ester synthesis in cultured human fibroblasts. N-SMase exerted a concentration-dependent continuous stimulation of 125I-LDL cell-surface binding, internalization, and degradation in normal human fibroblasts. A 3-fold increase in binding, internalization, and degradation was observed at the maximum amount (600 units of N-SMase/ml) examined. This phenomenon was accompanied by a continuous stimulation of cholesteryl ester synthesis. A 5-fold increase in cholesteryl ester synthesis was observed after incubation for 4 h with N-SMase. Antibody against N-SMase and heat inactivation of N-SMase compromised the stimulatory effects of N-SMase on 125I-LDL metabolism and cholesteryl ester synthesis in these cells. Incubation of cells with phospholipase D and phospholipase C did not alter 125I-LDL binding, internalization, or degradation. This finding suggests that the stimulatory effects of N-SMase on LDL metabolism and on cholesteryl ester synthesis in fibroblasts is specific. Moreover, unlabeled LDL competitively displaced 125I-LDL from binding to N-SMase-treated cells. None of the precursors of sphingomyelin could mimic the stimulatory effects of N-SMase on 125I-LDL metabolism in these cells. Taken together, these studies suggest that one of the biological roles of N-SMase involves modulating LDL metabolism and cholesterol metabolism in fibroblasts. PMID- 8429016 TI - Acid base catalytic mechanism of the dihydropyrimidine dehydrogenase from pH studies. AB - Primary deuterium (NADPH(D)), solvent deuterium, and multiple isotope effects and the pH dependence of kinetic parameters have been used to probe the mechanism of the dihydropyrimidine dehydrogenase from pig liver. Isotope effect and pH-rate data suggest a rate-determining reductive half-reaction in which reduction of the flavin by NADPH has only a minor rate limitation (DV approximately D(V/KNADPH) approximately 1.1), while protonation of the flavin at N-1 occurring in a step following reduction is slow (D2OV = 3, while D2O(V/KNADPH) = 2). An enzymatic general acid with a pK of 8.2 is required to protonate N-1 of the flavin. In the second half-reaction, uracil is reduced at C-6 by flavin and protonated on the opposite face at C-5 by an enzymatic general acid with a pK of 9. The hydride transfer from N-5 of the flavin to C-5 of uracil is facilitated by an enzymatic general base with a pK of 5.6 that accepts a proton from N-1 of the flavin. There is also evidence from the pH dependence of V and the V/K for reduced dinucleotide substrates that a second enzyme residue with a pK of 6.4 must be unprotonated for optimum activity, but is not essential for activity. None of the functional groups reflected in the V/KNADPH pH-rate profile have a role in binding, while both of those observed in the V/Kuracil profile have a role in binding as shown by the pH dependence of the dissociation constants for the competitive inhibitors ATP-ribose and 2,6-dihydroxypyridine. PMID- 8429017 TI - Functional organization of microtubule-associated protein tau. Identification of regions which affect microtubule growth, nucleation, and bundle formation in vitro. AB - Tau protein is a microtubule-associated protein that is almost exclusively expressed in the brain and is enriched in the axon. Determination of tau's sequence has revealed three to four tandem repeats that have been shown to constitute the microtubule binding site. In order to study the functional organization of tau, we prepared a series of truncated tau fragments using an Escherichia coli expression system. We assayed each fragment's activity in promoting growth of microtubules and in nucleating free microtubules. We found that tau's ability to nucleate microtubules requires the presence of additional sequence amino-terminal to that required for growth. We demonstrate that tau's carboxyl and amino termini differentially affect microtubule growth and nucleation. Finally, we show that in vitro microtubule bundle formation occurs when tubulin is assembled in the presence of an amino- and carboxyl-terminally truncated tau protein, whereas almost no bundling is observed in the presence of full-length tau or tau fragments that contain the amino terminus in addition to the repeat domain. We conclude that although the presence of the repeat domain promotes the growth of microtubules, the structural requirements for nucleation activity are more stringent. The differentiation between the growth promoting and nucleation activities on the structural level makes it possible for the two activities to be differentially regulated in vivo. PMID- 8429018 TI - Induction of Chinese hamster HSP27 gene expression in mouse cells confers resistance to heat shock. HSP27 stabilization of the microfilament organization. AB - Heat shock induces in cells the development of a transient state of thermotolerance thought to result from the induction of heat shock proteins. To assess directly whether a transient overexpression of one of these proteins, HSP27, can contribute to increased cellular resistance, mouse NIH/3T3 cells were cotransfected with a plasmid containing the Chinese hamster HSP27 gene under the control of the metallothionein promoter and a plasmid containing the neo gene. Stable transfectant cell lines were selected for resistance to the antibiotic G418. Analyses of several stable transfectant cell lines indicated that expression of Chinese hamster HSP27 could be selectively induced by exposure to 3 microM CdCl2, a concentration that had no effect on the induction of the endogenous heat shock proteins (HSP). In clone 15, the level of HSP27 increased steadily during the first day of exposure to CdCl2, from a concentration of 1 microgram/mg of total protein to 7 micrograms/mg. After withdrawal of CdCl2, the level of HSP27 returned to normal within the next 5 days. Accumulation of the Chinese hamster HSP27 was accompanied by a progressive development of thermoresistance that attained a level approaching heat shock-induced thermotolerance. After CdCl2 removal, thermal resistance and HSP27 decayed in a coordinated manner. In control cells transfected with the neo gene only, increased thermoresistance was not induced by 3 microM CdCl2; in these cells, an exposure to 20 microM CdCl2 was required to induce a level of thermoresistance comparable to that induced by 3 microM CdCl2 in clone 15. Elevated expression of HSP27 was accompanied by an increased stability of stress fibers during hyperthermia. The protein also partially prevented actin depolymerization during acute exposure to cytochalasin D and reduced cytotoxicity and growth inhibition of chronic exposures to the drug. The results indicated that accumulation of HSP27, as it occurs after a mild heat shock or other inducing treatments, is sufficient for acquisition of thermotolerance that may result in part from a stabilization of actin filaments. PMID- 8429019 TI - Protein-DNA interactions in the epsilon-globin gene silencer. AB - The developmental control of expression of the human epsilon-globin gene appears to be mediated, at least in part, by a transcriptional silencer in the DNA 5' to the cap site of this gene. We have used site-directed mutagenesis and DNA-protein binding assays to define the active motifs of this epsilon-globin silencer. DNase I foot-printing of the silencer region with K562 cell nuclear extracts defined a sequence, which we designate as the epsilon-globin silencer motif or epsilon GSM (epsilon -278 to -258 base pairs (bp)) containing a region (epsilon -270 to -258) with 90% homology to the yeast mating type silencer, ABF-1 (autonomous replicating sequence binding factor one) and which also overlaps at (epsilon -269 to -262) with the human YY1 consensus sequence, an element which mediates transcription repression and activation of viral, mouse, and human genes. The DNase I footprint extended 5' in the silencer region to include an inverted repeat of a six-nucleotide motif (epsilon -267 to -278 bp) which shares 5 of 6 bases with the GATA-1 consensus sequence. In gel mobility shift assays, two specific proteins (A and B) in nuclear extracts from erythroleukemia K562 cells bound to the DNase I-footprinted region. Protein B, associated with epsilon globin silencer activity in vitro, required an intact epsilon GSM sequence for binding. Mutation of 5 bases within the epsilon GSM in an epsilon-globin promoter containing fragment extending upstream to 1400 bp in transient transfection assays increased activity by 3.0-fold compared with the native sequence, suggesting that the silencer activity was mediated by the epsilon GSM sequence. We found that protein A could be displaced by a competitor containing the GATA-1 consensus sequence, suggesting that protein A is a GATA-like protein. The region from -267 to -271 within the epsilon GSM and GATA-1 homology region was important for binding of both proteins A and B. These data suggest that protein binding to the epsilon GSM and GATA motifs mediate the negative effect of the silencer on transcription, possibly via direct competition for binding to this DNA region. Recombinant yeast ABF-1 and human YY1 bound to the epsilon GSM. Mutating three bases (epsilon -259, -262, -264) in the epsilon GSM decreased the binding affinity of protein B and recombinant human YY1 but increased the binding affinity of recombinant yeast ABF-1. Furthermore, competitor containing the YY1 consensus sequence competed for protein B binding, whereas competitor containing a perfect yeast ABF-1 consensus sequence did not.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429020 TI - Characterization of the gene encoding human platelet glycoprotein IX. AB - Glycoprotein IX is a relatively small (M(r) 20,000) surface glycoprotein of human platelets; one of three (Ib alpha, Ib beta, IX) polypeptide chains present in the glycoprotein Ib-IX complex that functions as the von Willebrand factor receptor and mediates platelet adhesion in the arterial circulation. Using a cDNA for human glycoprotein IX as the probe, clones were isolated from a human genomic library, and the genomic sequence for glycoprotein IX (3.2 kilobases) was determined. The transcriptional start site was located by RNase protection and primer extension experiments. The gene includes three exons and two introns within 1.6 kilobases of DNA, and the entire open reading frame for glycoprotein IX is included within the third exon. The genes for glycoproteins IX and Ib alpha share similar exon sequences on the 5' side of their ATG start codons, and both genes possess introns in this region. The glycoprotein IX gene contains two consensus regulatory sequences (GATA and ACTTCCT [ets]) in its promoter region (5' flank, within 67 bases of the start site) that are also present in similar sites in the previously described "megakaryocyte-platelet" genes (glycoprotein IIb, platelet factor 4, beta-thromboglobulin: human and rat). Thus, the glycoprotein IX gene shares structural features with other megakaryocyte-platelet genes and contains at least two consensus cis-acting regulatory elements that may govern gene expression. PMID- 8429021 TI - GDF-3 and GDF-9: two new members of the transforming growth factor-beta superfamily containing a novel pattern of cysteines. AB - Two new mammalian members (growth/differentiation factor 3 (GDF-3) and GDF-9) of the transforming growth factor-beta superfamily were identified using degenerate oligonucleotides corresponding to conserved regions among known family members. By Northern analysis, GDF-3 transcripts were detected primarily in adult bone marrow, spleen, thymus, and adipose tissue. In contrast, GDF-9 transcripts were detected only in the ovary. Based on their cDNA sequences, the predicted GDF-3 and GDF-9 polypeptides each contain a potential signal sequence for secretion, a putative tetrabasic proteolytic processing site, and a COOH-terminal region that shows significant homology to the known members of the transforming growth factor beta superfamily. In the COOH-terminal region, GDF-3 and GDF-9 are most homologous to Xenopus Vg-1 (57%) and human bone morphogenetic protein 4 (34%), respectively. Unlike all previously described members of this superfamily, both GDF-3 and GDF-9 lack the conserved cysteine residue that is believed to form the sole disulfide linkage between subunits in other family members. These findings raise new possibilities regarding subunit interactions among members of this superfamily. PMID- 8429022 TI - Stimulation of calcium uptake in Saccharomyces cerevisiae by bovine protein kinase C alpha. AB - Ca2+ plays essential roles as a second messenger often in synergism with the calcium- and phospholipid-dependent phorbol ester receptor, protein kinase C (PKC), which stimulates Ca2+ influx in various cell types in a potential positive feedback mechanism. To address the compatibility of these mechanisms between lower eukaryotes and mammals, we have stably expressed bovine PKC alpha in the yeast Saccharomyces cerevisiae. We find that phorbol ester binding sites are created which stimulate a specific calcium- and phospholipid-dependent catalytic activity in vitro. Phorbol ester activation in vivo stimulates PKC down regulation, uptake of extracellular Ca2+, Ca2+ dependence of cell viability, and changes in cell morphology. This may represent activation of a putative PKC mediated signaling pathway utilized by functional yeast homologs of mammalian PKC isoforms. These are suggested by some protein data; however, their genes have not yet been characterized (Simon, A. J., Milner, Y., Saville, S. P., Dvir, A., Mochly-Rosen, D., and Orr, E. (1991) Proc. R. Soc. Lond. B 243, 165-171). Our findings indicate that bovine PKC alpha is functional in yeast and stimulates calcium uptake in a manner similar to some of its responses in mammalian cells, which suggests compatible aspects of higher and lower eukaryotic signaling pathways and the feasibility of dissecting parts of the action of common signaling mediators in a simple genetic model. PMID- 8429023 TI - The human endothelin-B receptor gene. Structural organization and chromosomal assignment. AB - The gene encoding the human endothelin-B receptor (hET-BR) has been isolated, and its structural organization and chromosomal assignment have been determined. Southern blot analysis demonstrated a single copy of the hET-BR gene in the human genome. The hET-BR gene spans 24 kilobases and consists of seven exons and six introns. The size range for exons is 109-2855 base pairs, although that for introns is 0.2-14.5 kilobases. Every intron occurs near the border of the putative transmembrane domain in the coding region. The major transcription initiation sites were mapped to the positions 258 and 229 base pairs upstream of the ATG initiation codon by primer extension and nuclease S1 protection experiments. The 5'-flanking region of the hET-BR gene lacks conventional TATA and CCAAT boxes but contains a sequence of potential Sp1 binding sites upstream of the transcription initiation sites. There are some canonical consensus sequences of cis-elements including GATA motif, acute phase reactant regulatory element, and E box. Using human-rodent somatic hybrid cell lines, the hET-BR gene was assigned to human chromosome 13. The present study will lead to a better understanding of the mechanism for the transcriptional regulation of the hET-BR gene and will give a clue as to how to search for possible genetic disorders of hET-BR. PMID- 8429024 TI - Analysis of Gz alpha by site-directed mutagenesis. Sites and specificity of protein kinase C-dependent phosphorylation. AB - The G protein alpha subunit Gz alpha is a substrate for phosphorylation by protein kinase C. The phosphorylation has been documented both in human platelets and in vitro and is characterized by a high degree of selectivity in relation to other G protein alpha subunits. We have demonstrated previously by phosphoamino acid analysis and CNBr peptide mapping that phosphorylation occurs at a serine residue(s) within the NH2-terminal 53 residues of Gz alpha. In this study, we have examined the site of phosphorylation using site-directed mutagenesis. Gz alpha variants containing selected substitutions of alanine for serine residues were expressed in human kidney 293 cells, and the ability of each to be phosphorylated in response to phorbol 12-myristate 13-acetate was examined. A focus was placed on Ser25 and Ser27, the 2 serine residues within a sequence of Gz alpha used to obtain a phosphorylation-sensitive antibody. The results demonstrate that Ser27 is the primary site of phosphorylation. Conversion of Ser27 to an alanine resulted in a 65% decrease in incorporation of [32P] phosphate; conversion of Ser25 had no effect. Conversion of Ser16, which like Ser25 and Ser27 conforms to a consensus site for protein kinase C, resulted in a modest (15%) decrease. The conversion of both Ser16 and Ser27 resulted in an 80% suppression of incorporation. In addition to these results, we have extended studies of the subunit and kinase selectivity of phosphorylation in platelets. We show here that under conditions promoting phorbol 12-myristate 13-acetate stimulated phosphorylation of Gz alpha in permeabilized platelets, Gq alpha is not phosphorylated. Moreover, Gi alpha, Gz alpha, and Gq alpha were not phosphorylated in response to analogues of cAMP or cGMP. Thus, only Gz alpha is phosphorylated in platelets and only in response to activation of protein kinase C. PMID- 8429025 TI - Characterization of a DNA mismatch-binding activity in yeast extracts. AB - An activity present in nuclear extracts of the yeast Saccharomyces cerevisiae binds specifically to oligonucleotides containing DNA mismatches, as judged by a band shift assay. The specificity of this activity for mismatched DNA was confirmed by competition experiments; binding to radiolabeled heteroduplexes was abolished in the presence of excess unlabeled heteroduplex but not when excess unlabeled homoduplex was added. Both T/G and T/- (single base deletion) mispairs were recognized in each of two sequence contexts. Binding was also observed with G/G, G/A, A/C, and T/C mismatches, but recognition of a C/C mispair was very weak. Competition studies with the various mismatches were consistent with the idea that a single activity recognizes all mispairs tested. Extracts from strains mutant in either or both of two putative mismatch recognition functions, MSH2 and MSH3, were also tested. Mismatch-binding activity was present in extracts of msh3 strains but completely absent in msh2- strains. The molecular weight of the major binding protein was estimated by UV cross-linking experiments to be approximately 110 kDa, in good agreement with the size predicted for Msh2 protein (Reenan, R. A. and Kolodner, R. D. (1992) Genetics 132, 963-973). PMID- 8429026 TI - The role of proline 345 in diphtheria toxin translocation. AB - Diphtheria toxin (DT) can translocate across endosomal membranes in response to low pH. Buried hydrophobic domains localized in the 37-kDa toxin B chain become exposed in response to acidic conditions and are thought to participate in the membrane translocation process. The crystal structure of DT has revealed a structurally distinct translocation domain composed of nine alpha-helices with their interconnecting loops (Choe, S., Bennett, M., Fujii, G., Curmi, P., Kantardjieff, K., Collier, R., and Eisenberg, D. (1992) Nature 357, 216-222). Two of these alpha-helices, TH8 and TH9, are unusually apolar and constitute the central core of the translocation domain. It has been proposed that these domains and the highly charged interconnecting loop undergo a conformation change under acidic conditions producing a dagger-like structure capable of inserting into the membrane thus initiating the translocation process. Proline 345 occupies a strategic location at the end of the TH8 alpha-helix. Proline residues have the ability to undergo a cis-trans isomerization reaction and because of this have been proposed to play a role in the conformational change that is a prerequisite for toxin translocation. The role of the proline at position 345 in membrane translocation was investigated. Pro was mutagenized to Glu and to Gly using a two step recombinant polymerase chain reaction procedure, and the mutant proteins were expressed in vitro. Glu, an alpha-helix former, and Gly, an alpha-helix breaker, were selected for mutagenesis to distinguish between a structural role for Pro as an alpha-helix breaker and alternative roles, perhaps involving cis trans isomerization-related conformational changes. Replacing Pro at position 345 with Glu or Gly resulted in a 99% reduction in toxicity to Vero cells. The enzymatic and binding activity of the toxin were not altered by the mutations. Instead, the reduction in toxicity is due to decreased translocation ability, suggesting that the Pro at position 345 plays a specific role in toxin membrane translocation. PMID- 8429027 TI - Protein kinase C is activated in platelets subjected to pathological shear stress. AB - High levels of fluid shear stress at the blood vessel wall directly stimulate von Willebrand factor (vWF)-mediated platelet adhesion and aggregation and thereby contribute to the pathogenesis of arterial thrombosis. We have found that a pathological level of arterial wall shear stress (90 dynes/cm2) induces platelet aggregation that is associated with the phosphorylation of pleckstrin, a M(r) 47,000 protein kinase C substrate (p47). Shear-induced p47 phosphorylation depends entirely on vWF binding to platelet glycoprotein (Gp) Ib and GpIIb-IIIa, and the specific inhibition of protein kinase C with the staurosporine analogue Ro 31-7549 inhibits the full aggregation response to shear. Shear stress-induced platelet p47 phosphorylation occurs independent of any measurable change in diacylglycerol mass or hydrolysis of phosphatidylinositol 4,5-bisphosphate. These results indicate that mechanical shear stress induces vWF to bind to platelet GpIb and GpIIb-IIIa, stimulating a diacylglycerol-independent pathway of protein kinase C activation that contributes to platelet aggregation in response to shear. PMID- 8429028 TI - Asparaginyl endopeptidase of jack bean seeds. Purification, characterization, and high utility in protein sequence analysis. AB - Asparaginyl endopeptidase was highly purified from mature seeds of the jack bean (Canavalia ensiformis). The final enzyme preparation showed a single peak in high performance liquid chromatography on a reversed-phase column, and the material in the peak gave the following NH2-terminal amino acid sequence on Edman degradation for 25 cycles: H-Glu-Val-Gly-Thr-Arg-Trp-Ala-Val-Leu-Val-Ala-Gly-Ser-Asn-Gly-Tyr Gly-Asn-Tyr- Arg-His-Gln-Ala-Asp-Val-. Behavior of the enzyme toward various protease inhibitors suggested that it belongs to a family of cysteine proteases. Strict substrate specificity of this enzyme was verified by the use of 14 polypeptide substrates including those derived from proteins. Almost all the peptide bonds on the carboxyl side of Asn residues were susceptible to the enzyme. The exceptions were cases where the residue was at the NH2 terminus or the second position from the NH2 terminus of substrates and where it was N glycosylated Asn. Peptide bonds on the carboxyl side of any other amino acid residues were not cleaved. These properties promise the high utility of this novel endopeptidase in protein sequence analysis. Identity of jack bean asparaginyl endopeptidase with a processing enzyme responsible for maturation of concanavalin A from its precursor is also discussed. PMID- 8429029 TI - The collagenous domains of macrophage scavenger receptors and complement component C1q mediate their similar, but not identical, binding specificities for polyanionic ligands. AB - Macrophage scavenger receptors have been implicated in the development of atherosclerosis and other macrophage-associated functions, including host defense. The mechanism by which these receptors bind a wide array of polyanions, such as acetylated low density lipoprotein (Ac-LDL), with high affinity has not yet been elucidated; however, it has been proposed that the positively charged extracellular collagenous domain of scavenger receptors plays a key role in ligand binding. To test this proposal, we generated truncation mutants of the bovine and murine scavenger receptors and studied their expression in transiently transfected COS cells. These mutants contain only 8 (bovine) or 5 (murine) of the 24 Gly-X-Y tripeptide repeats found in the collagenous domains of the full-length receptors. Immunochemical analyses established that the truncation of the bovine scavenger receptor did not interfere significantly with its synthesis, trimerization, post-translational processing, intracellular transport, surface expression, or stability. However, unlike their full-length counterparts, the truncated bovine and murine receptors were unable to bind Ac-LDL. Thus, the collagenous domain was necessary for normal ligand binding. In addition, cotransfection of the expression vector for the truncated bovine scavenger receptor with that for the full-length receptor resulted in dramatically reduced activity of the full-length construct (dominant negative effect). A ligand bead binding assay was used to show that the isolated collagenous domain from a different protein, complement component C1q, could bind a wide variety of polyanions with a specificity which was similar, but not identical, to that of scavenger receptors. These results suggest that the collagenous domain of the scavenger receptor is both necessary and sufficient to determine the broad binding specificity that characterizes this unusual receptor. Scavenger receptors and C1q, along with the mannose-binding protein, conglutinin, and lung surfactant apoprotein A, help define a set of proteins which all contain short collagenous domains and which all appear to participate in host defense. Their short collagenous domains may contribute significantly to their host-defense functions. PMID- 8429030 TI - Polynucleotide binding to macrophage scavenger receptors depends on the formation of base-quartet-stabilized four-stranded helices. AB - Macrophage scavenger receptors exhibit unusually broad, but circumscribed, polyanionic ligand-binding specificity. For example, the polyribonucleotides poly(I) and poly(G) are ligands but poly(A) and poly(C) are not. To further investigate the molecular basis of this polynucleotide-binding specificity, we tested the capacity of various oligodeoxyribonucleic acids to inhibit the scavenger receptor-mediated degradation of 125I-labeled acetylated low density lipoprotein by Chinese hamster ovary cells expressing the type I bovine scavenger receptor. A series of short oligodeoxyriboguanines (dGn, where 5 < or = n < or = 37) were effective inhibitors. The dG6, dG12, and dA5G37 members of this series were shown by circular dichroism and UV spectroscopy to be assembled into four stranded helices stabilized by G-quartets. [32P]dA5G37 bound directly to scavenger receptors. Partial or complete denaturation of the quadruplex structures of these oligonucleotides by boiling destroyed their inhibitory activity. Receptor activity was also inhibited by d(T4G4)4, a telomere-like oligonucleotide which forms an intramolecular quadruplex. In addition, conversion of the four-stranded potassium salt of poly(I) to the single-stranded lithium salt dramatically reduced its inhibitory activity. Addition of KCl to the Li+ salt resulted in the reformation of poly(I)'s quadruplex structure and restoration of its inhibitory activity. A variety of single-stranded and double stranded oligo- and polydeoxyribonucleotides (e.g. dA37, HaeIII restriction fragments of phi X174) exhibited very little or no inhibitory activity. Thus, a base-quartet-stabilized four-stranded helix appears to be a necessary structural determinant for polynucleotide binding to and inhibition of scavenger receptors. This conformational requirement accounts for the previously unexplained polyribonucleotide-binding specificity of scavenger receptors. The spatial distribution of the negatively charged phosphates in polynucleotide quadruplexes may form a charged surface which is complementary to the positively charged surface of the collagenous ligand-binding domain of the scavenger receptor. PMID- 8429031 TI - Assembly of rat hepatic very low density lipoproteins in the endoplasmic reticulum. AB - The intracellular site of assembly of hepatic very low density lipoproteins has been investigated. Two endoplasmic reticulum fractions and Golgi vesicles (relatively free from endosomal contamination) were isolated from rat liver and the luminal contents were released. The apoB-containing entities were separated from the lumen of the endoplasmic reticulum and Golgi vesicles by an immunoaffinity isolation procedure. The amount of each lipid moiety (triacylglycerols, cholesterol plus cholesteryl esters and phospholipids) associated with a unit mass of apoB was shown to be very similar in the luminal contents isolated from each of the three fractions. Moreover, the apoB-containing particles that were isolated from the endoplasmic reticulum and Golgi were mainly present in a fraction of density < 1.02 g/ml. The average diameter of these lipoprotein particles was shown by negative staining electron microscopy to be in the size range of very low density lipoproteins and low density lipoproteins. The size and composition of the intracellular lipoproteins were very similar to those of both very low density lipoproteins isolated from the culture medium from rat hepatocytes and plasma very low density lipoproteins. The conclusion from this study is that the full complement of lipids associate with apoB at an early stage during the secretory pathway, in the endoplasmic reticulum. PMID- 8429032 TI - Promoter elements and transcriptional control of the mouse acetylcholinesterase gene. AB - The 5'-untranslated region of the mouse acetylcholinesterase gene has been characterized structurally by RNase protection, primer extension, and sequencing. Evidence has been obtained for the use of two alternative promoters in brain. Tissue-specific splicing to alternative acceptor sites in the 5'-untranslated exons occurs in brain, muscle, and erythropoietic cells. cis elements 5' of the cap site that is predominantly used in these tissues and cells have been analyzed by deletion analysis of promoter-reporter gene constructs and by site-specific mutagenesis. The cap site is found 107 base pairs (bp) 5' of the translation start site. This region is devoid of CAAT or TATA sequences; further in the 5' direction 50 and 70 bp are tandem Egr-1 sites. The putative promoter has been coupled to the open reading frame of a luciferase reporter gene. Deletion analysis shows that this region largely accounts for tissue-specific transcription seen upon transfection of neuronal and muscle cells. Mutagenesis of the Egr-1 sites results in a marked loss of reporter gene activity, further substantiating the importance of this region in the control of transcription. cis elements in the promoter differ from those found for the genes encoding the various subunits of the nicotinic acetylcholine receptor, and distinct differences in control of transcription are evident when the respective reporter genes are transfected into C2 muscle cells. PMID- 8429033 TI - Growth hormone-promoted tyrosyl phosphorylation of a 121-kDa growth hormone receptor-associated protein. AB - Previous work in multiple cell types has shown that endogenous GH receptors, as well as the cloned liver GH receptor, associate with a tyrosine kinase. However, in SDS-PAGE gels of highly purified, kinase-active GH receptor preparations from 35S-labeled 3T3-F442A cells, only one broad band was detected corresponding to the molecular weight of the GH receptor rather than two bands which might be expected to result from a kinase-receptor heterocomplex. In the present study, a transfected Chinese hamster ovary (CHO) cell line (CHO4) that expresses an 84-kDa GH receptor rather than a 121-kDa GH receptor was used to examine whether the GH receptor might form a complex with a protein (e.g. tyrosine kinase) that comigrates on SDS-polyacrylamide gel electrophoresis gels with the endogenous GH receptor (M(r) 121,000) in 3T3-F442A cells. GH-GH receptor complexes were immunoprecipitated with anti-GH antibody from GH-treated CHO4 cells and incubated with [gamma-32P]ATP. 32P was incorporated into a 121-kDa protein as well as the 84-kDa GH receptor. Phosphorylation of both the 84-kDa GH receptor and the 121 kDa protein was on tyrosyl residues as determined by Western blotting with anti phosphotyrosine antibody. The 121-kDa protein does not appear to bind GH. It was also not detected in the immunoprecipitate when cells had not been incubated with GH or when untransfected CHO cells were used. These findings suggest that in CHO4 cells, the 121-kDa protein is precipitated by the GH antibody because of its ability to form a complex with the GH receptor (p84). Western blot analysis of whole cell lysates using anti-phosphotyrosine antibody revealed that GH promotes the tyrosyl phosphorylation of a 121-kDa protein and several other proteins (p97, p42, p39) in a dose- and time-dependent fashion. Taken together, these findings are consistent with either p121 being the tyrosine kinase that complexes with the GH receptor and is activated in response to GH binding or with p121 forming a ternary complex with both the GH receptor and a tyrosine kinase and serving as a substrate of the GH receptor-associated tyrosine kinase. PMID- 8429034 TI - Determinants of the intracellular fate of truncated forms of the platelet glycoproteins IIb and IIIa. AB - The platelet glycoproteins GPIIb and GPIIIa are integral membrane proteins and form calcium-dependent heterodimers in the endoplasmic reticulum (ER). In the absence of heterodimer formation, GPIIb and GPIIIa are retained in the ER and degraded. To produce soluble forms of these proteins, we truncated each at a site just proximal to its transmembrane anchor and expressed the mutants in COS-1 cells. We found that both truncated GPIIIa (GPIIIatr) and GPIIIatr were secreted by the transfected cells. However, GPIIbtr was retained by the cells and was immunoprecipitated as a doublet with a 115,000 molecular weight protein. Incubation of transfected cells with the calcium ionophore A23187 or the calcium chelator 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester (BAPTA-AM) failed to induce appreciable GPIIbtr secretion, suggesting that formation of intracellular calcium complexes was not a factor in GPIIbtr retention. Further, immunoblotting of immunoprecipitated GPIIbtr and GPIIIatr revealed that the chaperone binding protein (BiP) was associated with each, arguing that BiP alone was not responsible for GPIIbtr retention. These studies indicate that the intracellular retention of GPIIIa involves sequences located in the transmembrane or cytoplasmic domains of the molecule. GPIIb contains an additional retention signal located in the extracellular portion of the molecule whose effect is abrogated by formation of a GPIIb-IIIa heterodimer. This signal may be involved in the fate of nascent GPIIb monomers and the generation of correctly configured GPIIb-IIIa heterodimers. PMID- 8429035 TI - Translocation of conjugated presecretory proteins possessing an internal non peptide domain into everted membrane vesicles in Escherichia coli. AB - Polypeptides comprising 20 amino acid residues (Y2) were covalently bound to the carboxyl terminus of a truncated proOmpA (proOmpA-D72C) through N,N'-bis(3 maleimidopropionyl)-2-hydroxy-1,3-propanediamine (X). The length of the inverted linker domain was 2.8 nm. proOmpA-D72C-X-Y2 thus synthesized was subjected to in vitro translocation into everted membrane vesicles of Escherichia coli. The conjugated protein was translocation-competent in terms of both proteinase K resistance and signal peptide cleavage, when a proton motive force (delta microH+) was imposed. The translocation was ATP-dependent. The proteinase K treatment resulted in the digestion of SecA, SecE, and SecY in the membrane, suggesting that the proteinase K resistance of the Y2 domain was not due to its interaction with these Sec proteins in the secretory machinery. In the absence of delta microH+, the translocation ceased at the linker domain. Upon the imposition of delta microH+, the linker-Y2 domain underwent translocation, which did not require ATP hydrolysis as in the case of the translocation of the latter portion of usual secretory proteins. The translocation was prevented by anti-Y2 IgG even when delta microH+ was imposed. Another conjugated protein, which possesses a polypeptide comprising 61 amino acid residues after the linker (proOmpA-D72C-X Lpp'), was synthesized. This compound was also translocated into everted membrane vesicles with cleavage of the signal peptide. These results suggest that substances to be translocated through the secretory machinery need not necessarily be solely held together by polypeptide bonds. PMID- 8429036 TI - Repression and redirection of Saccharomyces cerevisiae tRNA synthesis from upstream of the transcriptional start site. AB - Derivatives of the Saccharomyces cerevisiae SUP4 tRNATyr gene with binding sites for the transcription regulatory protein GCN4 located upstream of the transcriptional start site have been constructed. The effect of GCN4 on transcription of these genes by purified RNA polymerase III and transcription factors (TF) IIIB and IIIC has been analyzed. GCN4 effectively blocks initiation of transcription only when prebound to sites that overlap with the binding site of TFIIIB. Residual GCN4-repressed transcription is significantly redirected to nearby downstream sites, the selection of which depends on the location of bound GCN4. That prebound repressing GCN4 redirects, instead of merely blocking, the TFIIIC-dependent interaction of TFIIIB with DNA has been directly demonstrated by footprinting. The effect of GCN4 on transcription persists after it has been stripped off its DNA-binding site: once it has been redirected, DNA-bound TFIIIB remains in place, a consequence of the fact that it binds extraordinarily tightly to DNA without recognizing specific DNA sequence. PMID- 8429037 TI - Thyrotropin regulation of sialic acid expression in rat thyroid cells. AB - The present study, utilizing the hormone-responsive rat thyroid cell line FRTL-5, presents evidence establishing the regulatory role of thyrotropin in modulating mRNA for beta-galactoside alpha 2,6-sialytransferase, the enzyme responsible for the expression of alpha 2,6-linked sialic acid. Both the cell surface membrane and the thyroglobulin secreted by cells grown in the presence of this hormone exhibit a marked decrease in the level of alpha 2,6-bound sialic acid with little or no change in the number of alpha 2,3-sialic acid residues. An additional, and unexpected, sequel is the finding of a coordinated decrease in all of the core monosaccharide constituents of the secreted thyroglobulin. Both of the above phenomenological changes appear to be at some variance with previously described systems wherein thyrotropin was deemed to increase glycosylation. It is anticipated that further resolution of this apparent difference may provide a clearer definition for the role of the carbohydrate moiety in affecting the biological function of thyroglobulin. PMID- 8429038 TI - The P1 plasmid partition complex at parS. II. Analysis of ParB protein binding activity and specificity. AB - The P1 plasmid prophage is partitioned by a very high affinity protein complex at its partition site, parS, that contains the P1 ParB protein and Escherichia coli integration host factor (IHF). ParB binds to regions of parS that flank the IHF binding site. In this report, we have examined the sequences to which ParB binds, the spatial relationship between them, and the effect of IHF on ParB binding patterns. Methylation protection and interference experiments were performed on supercoiled plasmids. Mutations that interfered with the action of both proteins in vivo were identified following random mutagenesis of parS. These studies revealed that ParB binds to a complicated, nonsymmetrical region in the right side of parS. ParB recognizes a partial copy of this sequence, TCGCCA, in the left side of parS with much lower affinity. The presence of IHF greatly facilitates the interaction of ParB with parS such that both sides bind with an equal affinity that is much greater than to either side alone. The stimulation by IHF is strongly influenced by helical phasing. These observations support the proposal that ParB is directed, by the bend created by IHF, to bind simultaneously to properly placed sequences flanking the IHF site. PMID- 8429039 TI - Interaction between apolipoproteins A-I and A-II and the membrane attack complex of complement. Affinity of the apoproteins for polymeric C9. AB - We have previously observed enhanced binding of HDL and apolipoproteins A-I and A II to human endothelial cells exposed to activated complement. Induction of these binding sites required complement activation through C9, suggesting a specific role for the C9 component of the C5b-9 complex. We now report that specific and saturable binding sites for apoA-I and -A-II are expressed by C9 polymers (polyC9), whereas little binding was observed to native monomeric C9. These data suggested an interaction of the apoproteins with a site(s) which is exposed only upon C9 polymerization, and also suggested that binding of the apoproteins to this new site might interfere with assembly of C9 into the polyC9 tubule and insertion into the cell membrane. ApoA-I was found to inhibit zinc-catalyzed polymerization of C9 in a concentration-dependent fashion. Formation of SDS resistant C9 polymers was completely inhibited at apoA-I or -A-II concentrations > or = 5 microM. ApoA-I also produced a concentration-dependent inhibition of C9 incorporation into C5b-9 complexes on endothelial cells, which was accompanied by a corresponding decrease in SDS-resistant C9 polymers associated with the cell membrane. In summary, the ability of the HDL apoproteins A-I and A-II to interact with an activation-dependent conformer(s) of the C9 component of the C5b-9 complex appears to explain the expression of HDL binding sites on endothelial cells exposed to complement. These apoproteins are also inhibitors of C9 polymerization, which may underlie the protective effect of HDL for blood cells exposed to activated complement. PMID- 8429040 TI - Mutagenesis of thrombin selectively modulates inhibition by serpins heparin cofactor II and antithrombin III. Interaction with the anion-binding exosite determines heparin cofactor II specificity. AB - Thrombin is a multifunctional serine protease that plays a critical role in hemostasis. Thrombin is inhibited by the serpins antithrombin III and heparin cofactor II in a reaction that is dramatically accelerated by glycosaminoglycans. The structural basis of the interaction with these inhibitors was investigated by introducing single amino acid substitutions into the anion-binding exosite (R68E, R70E) and unique insertion loops (K52E, K154A) of thrombin. The rate of inhibition of these recombinant thrombins by antithrombin III and heparin cofactor II was determined in the absence and presence of glycosaminoglycan. The second order rate constant (k2) for inhibition by antithrombin III without heparin was 3.7 x 10(5) M-1 min-1 for wild-type thrombin; rates for the mutant thrombins varied less than 2-fold. For inhibition by antithrombin III with heparin, the rate constant was 4.5 x 10(8) M-1 min-1 for wild-type thrombin with no significant differences between any of the recombinant thrombins. In contrast, the rate constant for inhibition by heparin cofactor II without glycosaminoglycan was 4.3 x 10(4) M-1 min-1 for wild-type thrombin; rates were 10-fold slower for thrombin K52E and 2- to 3-fold slower for thrombins R68E and R70E. The rate constants for inhibition of wild-type thrombin by HCII in the presence of heparin or dermatan sulfate were 9.2 x 10(8) M-1 min-1 and 9.0 x 10(8) M-1 min-1, respectively. Compared to wild-type thrombin, the rate of inhibition by HCII with glycosaminoglycan was 5- to 15-fold slower for thrombins K52E and R70E and 50- to over 100-fold slower for thrombin R68E. Thrombin K154A was inhibited by heparin cofactor II with rates similar to wild-type thrombin in all assays. These results suggest that heparin cofactor II interacts with residue Lys-52 in the proposed S1' subsite and with residues Arg-68 and Arg-70 in the anion-binding exosite of thrombin, and that these interactions contribute to the molecular basis of heparin cofactor II specificity for thrombin. PMID- 8429041 TI - Glucose catabolism in African trypanosomes. Evidence that the terminal step is catalyzed by a pyruvate transporter capable of facilitating uptake of toxic analogs. AB - The protozoan parasite Trypanosoma brucei derives its metabolic energy exclusively from a unique type of glycolysis in which pyruvate derived from glucose catabolism is released into the host bloodstream. In this study, this terminal metabolic step has been examined in detail. Pyruvate release from trypanosomal cells supplied with glucose is very rapid, proceeding with an apparent Vmax of 214 nmol x min-1 x mg-1. Counterflow experiments with [14C]pyruvate demonstrate that this metabolic end product can be taken up by actively metabolizing cells consistent with the presence of a plasma membrane transporter. The findings that [14C] acetate exhibits a much lower capacity for cell entry and that the structural analog alpha-cyano-3-hydroxycinnamic acid inhibits pyruvate release provide additional support for the presence of a pyruvate transporter. The substrate analog and alkylating agent 3-bromopyruvate inhibits completely both cell motility and pyruvate release. Surprisingly, however, it is a poor inhibitor of pyruvate transport per se. Rather, its preferential site of action and that of iodoacetic acid were identified by radiolabeling studies and microsequence analysis as glyceraldehyde-3-phosphate dehydrogenase. In extending these studies, 3-bromopyruvate was found to be over 20 times less effective in inhibiting glyceraldehyde-3-phosphate dehydrogenase in intact erythrocytes than in trypanosomal cells. However, in sonicated preparations from both cell types, the enzyme exhibits nearly identical sensitivities to inhibition by 3-bromopyruvate. Experiments reported here provide the first direct evidence that pyruvate release in African trypanosomes is catalyzed by a specific transport system and implicate this transporter as a vehicle for delivering toxic alkylating agents into trypanosomal cells. PMID- 8429042 TI - DNA sequence requirements for interaction of the RK2 replication initiation protein with plasmid origin repeats. AB - Replication of plasmid RK2 in a variety of Gram-negative bacteria requires its origin of replication and the plasmid-encoded TrfA proteins (TrfA-33 and TrfA 44). The initiation of replication requires that the TrfA proteins bind to a series of 17-base pair (bp) direct repeats located within the RK2 origin. The conserved 17-bp repeats are arranged in tandem and are separated by less conserved spacer sequences of 4-6 bp in length. A series of plasmids containing one or two iterons, with or without the less conserved spacer sequences, were constructed to analyze the DNA sequence requirements for binding of TrfA-33 to the iterons. In addition to the analysis of TrfA binding in vitro, the plasmid constructs were examined for their ability to exert incompatibility toward an RK2 replicon in Escherichia coli. These analyses revealed that the conserved 17-bp iteron sequence itself is not sufficient for TrfA binding; the adjacent less conserved spacer sequences are also required. Site-specific mutagenesis was carried out to determine the importance of specific bases within the spacer sequence for binding activity and a consensus sequence for a TrfA-specific binding unit was determined. DNase I and methylation interference footprinting procedures were also carried out to characterize the TrfA-binding unit complex. Finally, it was shown that the binding of the TrfA-33 protein to two adjacent TrfA binding units on a DNA fragment is not substantially affected by the relative orientation or spacing between the two units. PMID- 8429043 TI - Brefeldin A inhibits degradation as well as production and secretion of collagen in human lung fibroblasts. AB - Brefeldin A (BFA) inhibits protein secretion, collapses the Golgi complex into the endoplasmic reticulum (ER), causes redistribution of processing enzymes normally resident in the Golgi to the ER, and uncouples the proximal and distal regions of the secretory pathway. We used BFA to determine where intracellular degradation of newly synthesized collagen degradation occurs. In normal human fetal lung fibroblasts, BFA (50 ng/ml) completely blocked collagen secretion and reduced collagen production by two-thirds. In cells synthesizing collagen under normal conditions, intracellular degradation was about 16%; BFA (50 ng/ml) reduced degradation to less than 5%. In cells induced to synthesize structurally abnormal collagen (by incubation with the proline analog cis-hydroxyproline), degradation was approximately 33%; BFA reduced this level to less than 10%. When the y axes were scaled appropriately, the dose-response curves for collagen degradation +/- cis-hydroxyproline versus BFA concentration coincided. A pulse chase experiment demonstrated that BFA did not inhibit hydroxylation of prolyl residues, a major posttranslational modification of collagen that occurs in the ER, and that inhibition of degradation was independent of inhibition of collagen synthesis. Immunofluorescence examination revealed that BFA redistributed Golgi glycoproteins to the ER. At the ultrastructural level, Golgi complex could not be found in fibroblasts exposed to BFA for 1 h; however, clusters of small vesicles were observed. A different structure, comprising one or two lamellae and resembling a partial Golgi complex, was observed in cells incubated with BFA for 6 h. This structure was adjacent to ER but far from the nucleus. In addition, the ER was devoid of ribosomes. The inhibition of intracellular collagen degradation by BFA indicates that collagen degradation does not occur in the ER. Rather, it suggests that collagen degradation occurs beyond the BFA block, perhaps in the trans-Golgi network. PMID- 8429044 TI - Role of Ser-238 and Lys-240 in the hydrolysis of third-generation cephalosporins by SHV-type beta-lactamases probed by site-directed mutagenesis and three dimensional modeling. AB - A growing number of extended spectrum SHV-type beta-lactamases capable of hydrolyzing third-generation cephalosporins such as cefotaxime and ceftazidime have been reported. These new enzymes differ by a few amino acids from SHV-1, an enzyme incapable of hydrolyzing these drugs. Two of these substitutions, Gly-238- >Ser and Glu-240-->Lys, are in a key beta-strand of the catalytic site of class A beta-lactamases. To understand the structural basis of these new activities, we first subcloned the DNA region coding for SHV-1 and SHV-2 and did site-directed mutagenesis to create two mutant SHV-1 proteins containing Ser and Glu or Gly and Lys and two mutant SHV-2 proteins containing Gly and Glu or Ser and Lys in positions 238 and 240, respectively. Phenotypic analysis (antibiograms and minimum inhibitory concentrations) and activity spectra of mutant enzymes showed that Ser-238 is critical for cefotaxime hydrolysis whereas both Ser-238 and Lys 240 are needed for strong ceftazidime hydrolysis. A three-dimensional model for SHV beta-lactamase complexes was constructed using the crystallographic structure of the homologous Bacillus licheniformis beta-lactamase, the complex of cefotaxime with the Streptomyces sp. R61 D-alanyl-D-alanine peptidase, and the complex of aztreonam with the Citrobacter freundii beta-lactamase. The modeling of SHV beta-lactamase complexes showed that factors which are most likely to correlate with binding and kinetic data are the size of the relatively buried amino acid at position 238 and the electrostatic charge of the exposed group at position 240. PMID- 8429045 TI - Removal of the carboxyl-terminal region of phospholipase C-beta 1 by calpain abolishes activation by G alpha q. AB - The 150-kDa phospholipase C (PLC)-beta 1 and three immunologically related proteins with molecular sizes of 140, 100, and 45 kDa were purified from bovine brain extracts. Determination of the amino-terminal amino acid sequence of the 45 kDa protein and immunoblots of the purified proteins with sequence-specific antibodies to peptides corresponding to three different regions of PLC-beta 1 suggest that a single cleavage at the linkage between amino acid residues 880 and 881 of PLC-beta 1 generates the 100- and 45-kDa proteins, which correspond to the amino-terminal and carboxyl-terminal portions, respectively, of PLC-beta 1. The Ca(2+)-dependent protease calpain appears to be responsible for the cleavage of PLC-beta 1; the PLC-beta 1 amino acid sequence contains PEST sequences which are common to proteins susceptible to calpain, and limited proteolysis of purified PLC-beta 1 by calpain generated a 100-kDa protein and a 40-kDa protein that contains the same amino-terminal sequence as the 45-kDa protein. The 140-kDa protein lacks the carboxyl-terminal-most region of PLC-beta 1, but there is no evidence it is derived from PLC-beta 1 by proteolysis. Cleavage of PLC-beta 1 by calpain had no significant effect on catalytic activity measured in the absence of the alpha subunit of the G alpha q but completely abrogated the stimulatory effect of G alpha q. On the other hand, G alpha q activated the 140-kDa enzyme. These results suggest that the region between residue 881 and the most carboxyl terminal 10 kDa of PLC-beta 1 contains the G alpha q interaction site. PMID- 8429046 TI - The regulatory domain of protein kinase C beta 1 contains phosphatidylserine- and phorbol ester-dependent calcium binding activity. AB - Certain isoforms of protein kinase C (PKC) require both Ca2+ and phospholipid for optimum activity. However, little is known about the nature of the interaction between PKC and Ca2+. The present study demonstrates that the isolated regulatory domain of PKC beta 1, when synthesized as a fusion protein in Escherichia coli, binds 45Ca2+ with high affinity, but only in the presence of phosphatidylserine or 12-O-tetradecanoyl-phorbol-13-acetate. This binding is highly selective for Ca2+ since it is preferentially inhibited by excess non-radioactive Ca2+ when compared with the cations Mg2+, Mn2+, Na+, or K+. It appears, therefore, that the binding of Ca2+ to PKC requires a complex tertiary structure in the regulatory domain. PMID- 8429047 TI - Regulation of protease nexin-1 synthesis and secretion in cultured brain cells by injury-related factors. AB - The clotting protease thrombin might contribute to cell damage following brain injury by its ability to retract processes on neurons and astrocytes. Protease nexin-1 (PN-1), a potent inhibitor of thrombin, is localized around cerebral blood vessels where it may protect these cells from extravasated thrombin during injury or alteration of the blood-brain barrier. Here we examined the effects of several injury-related factors on the regulation of PN-1 in cultured brain cells. Interleukin-1, tumor necrosis factor-alpha, and transforming growth factor-beta stimulated the secretion of PN-1 by the neuroblastoma cell line SK-N-SH. This cell line comprises both neuronal and glial cells. Analyses using cloned derivatives of these two cell types showed that PN-1 was secreted by the glial cells; PN-1 secretion was stimulated 90-fold by interleukin-1, 15-fold by tumor necrosis factor-alpha, 10-fold by tumor growth factor-beta, and 4-fold by platelet-derived growth factor. Measurements of newly synthesised PN-1 demonstrated that these factors produced an equivalent stimulation of PN-1 synthesis. The neuronal cells secreted two thrombin-binding proteins distinct from PN-1. Interactions between these two cell types regulated the secretion of PN-1 and the two thrombin-binding proteins. PMID- 8429048 TI - Rapamycin-induced inhibition of p34cdc2 kinase activation is associated with G1/S phase growth arrest in T lymphocytes. AB - The macrolide rapamycin (RAP) is a potent inhibitor of interleukin-2 (IL-2) induced T-cell proliferation. Current models suggest that RAP, when complexed to its intracellular receptor, FK506-binding protein, interferes with an IL-2 receptor-coupled signaling pathway required for cell-cycle progression from G1- to S-phase. Here we show that RAP treatment inhibits the growth of an IL-2 dependent cytotoxic T-cell line, CTLL-2, in late G1-phase, just prior to entry of the cells into S-phase. In contrast, RAP-treated CTLL-2 cells retained the ability to respond to IL-2 with enhanced cytolytic activity, indicating that RAP was not a general suppressant of cellular responsiveness to IL-2. Subsequent studies revealed that IL-2 stimulation triggered a delayed activation of the p34cdc2 kinase, the timing of which correlated with the G1- to S-phase transition. The IL-2-dependent increase in p34cdc2 kinase activity was blocked by RAP. The RAP sensitivity of the p34cdc2 activation mechanism implicates this signaling pathway in the control of S-phase commitment in IL-2-stimulated T cells. PMID- 8429049 TI - Epidermal growth factor-mediated signaling of G(i)-protein to activation of phospholipases in rat-cultured hepatocytes. AB - Hepatocytes were established in tissue culture in order to study the effects of pertussis toxin (PT) on epidermal growth factor (EGF)-mediated cellular responses under in vitro conditions. EGF caused a 3-fold increase of myo-inositol 1,4,5 trisphosphate (Ins-1,4,5-P3) mass and a 50% increase of diacylglycerol mass within the first minute, with the change of diacylglycerol content being 100-fold greater than that of Ins-1,4,5-P3. Diacylglycerol, but not Ins-1,4,5-P3, continued to accumulate over several hours, indicating that EGF increased the hydrolysis of lipids other than phosphatidylinositol 4,5-bisphosphate (PIP2). EGF increased phosphoinositide-specific phospholipase C-gamma (PLC-gamma) tyrosine phosphorylation within 1 min, but no effect was observed with vasopressin, insulin, or glucagon after 5 min. EGF also caused a rapid, tyrosine kinase dependent association of G(i) alpha with PLC-gamma, which was maximal within 10 min. In contrast to our previous data on fresh hepatocytes, PT had no effect on the EGF-induced tyrosine phosphorylation of PLC-gamma, although Ins-1,4,5-P3 and diacylglycerol production were inhibited. The role of G-proteins in EGF signaling was investigated further by microinjection of G alpha antibodies into single fura 2-loaded hepatocytes. Anti-G(i) alpha (common) antibodies prevented EGF-induced but not vasopressin-induced Ca2+ transients. These results strengthen previous observations that a PT-sensitive G-protein is involved in EGF-mediated phospholipid metabolism in hepatocytes and show that tyrosine phosphorylation of PLC-gamma is an insufficient signal for activation of PIP2 hydrolysis. PMID- 8429050 TI - Genomic structure of the locus encoding protein 4.1. Structural basis for complex combinational patterns of tissue-specific alternative RNA splicing. AB - Protein 4.1 (P4.1) is a multifunctional protein with heterogeneity in molecular weight, intracellular localization, tissue- and development-specific expression patterns. We have analyzed the genomic structure of the locus encoding mouse P4.1 and have systematically analyzed diverse P4.1 mRNA isoforms expressed in erythroid and nonerythroid tissues. Our results indicate that the mouse protein 4.1 gene, over 90 kilobases long, comprises at least 23 exons (13 constitutive exons, 10 alternative exons) interrupted by 22 introns. The donor and acceptor splice site sequences match the consensus sequences for the exon-intron boundaries of most eukaryotic genes. No significant sequence difference was observed between splice junctions of alternative and constitutive exons. Apparently, most alternative exon-encoded peptides are located within particular functional domains of the P4.1 protein: two peptides encoded by alternative exons 4 and 5 are located near or within the glycophorin/calmodulin binding domain, whereas three other alternative exon-encoded peptides (19-amino acid encoded by exon 14, 14-amino acid encoded by exon 15, and 21-amino acid encoded by exon 16) are located near or within the spectrin-actin binding domain. Selective use of exon 2', which carries an upstream translation initiation codon (AUG), may produce an elongated P4.1 isoform (135 kDa) that is predominantly expressed in nonerythroid tissues. Combinatorial splicing of these exons may generate different isoforms that exhibit complicated tissue-specific expression patterns. PMID- 8429051 TI - The effects of low density lipoprotein on calcium transients in isolated rabbit cardiomyocytes. AB - The purpose of this study was to examine the effects of low density lipoprotein (LDL) on Ca2+ transients of isolated rabbit cardiomyocytes. Incubation of cardiomyocytes with > or = 1 mg of LDL cholesterol/ml of perfusion medium induced a slow (> or = 30 min) but significant increase (2-fold) in the cellular Ca2+ transient. The time course for the effect was similar to that observed for the accumulation of cholesterol in the cells. Using Dil- labeled LDL as a fluorescent marker for LDL interaction with the cardiomyocytes, it was concluded that LDL interacted via a receptor-mediated event, but probably this was not the primary mechanism whereby the lipid entered the cell. LDL-treated cells were resistant to the depressant actions for ryanodine, nicardipine, and dichlorobenzamil on the cellular Ca2+ transient. Lowering the extracellular Ca2+ concentration removed the stimulatory effect of LDL on the Ca2+ transient. It is concluded that LDL can induce an increase in the magnitude of the Ca2+ transient in isolated cardiomyocytes. This is a relatively slow process. The mechanism appears to involve a stimulation of a transsarcolemmal Ca2+ transport pathway. These findings have important implications for cardiac contractile function in hypercholesterolemic and drug-treated hypercholesterolemic subjects. PMID- 8429052 TI - Regulation of actomyosin interactions in Limulus muscle proteins. AB - Contraction of striated muscle from Limulus polyphemus, the horseshoe crab, is regulated by both calcium binding to a troponin-tropomyosin-dependent thin filament array and a myosin light chain kinase-dependent phosphorylation of myosin. We have isolated myosin from Limulus striated muscle and examined how these two regulatory systems affect the sliding velocity of actin filaments over myosin, using an in vitro motility assay. Our results show that in the presence of ATP, Limulus myosin must be phosphorylated in order to move actin filaments. No movement was observed for actin filaments interacting with dephosphorylated Limulus myosin. Calcium was not required for actin movement. In contrast, when both troponin and tropomyosin are bound to actin filaments, calcium is required for the movement of actin filaments over phosphorylated myosin. These results demonstrate that the "off" state of either the thin filament or thick filament regulatory system is dominant and that for the movement to occur, both phosphorylated Limulus myosin and an activated troponin-tropomyosin system are required. Tropomyosin by itself increases the sliding velocity of actin filaments over phosphorylated Limulus myosin about 10-fold in a calcium-independent manner. Tropomyosins from turkey gizzard smooth muscle, bovine cardiac muscle, and Limulus muscle all have a profound effect in increasing the velocity. Troponin alone does not change the velocity. Partial sequences of the tryptic phosphopeptides of Limulus myosin regulatory light chains generated following the phosphorylation by gizzard myosin light chain kinase yield ATS(PO4)NVFAMFEQNQIA for 21 kDa and SGS(PO4)NVFSMFT for 31-kDa light chain. PMID- 8429053 TI - A neural network representation of electromyography and joint dynamics in human gait. AB - Optimization theory and other mathematical algorithms have traditionally been used to model the relationship between muscle activity and lower-limb dynamics during human gait. We introduce here an alternative approach, based on artificial neural networks with the back-propagation algorithm, to map two different transformations: (1) EMG-->joint angles; and (2) EMG-->joint moments. Normal data for 16 muscles and three joint moments and angles (hip, knee, and ankle) were adapted from the literature [Winter (1987), The Biomechanics and Motor Control of Human Gait]. Both networks were successfully trained to map the input vector onto the output vector. The models were tested by feeding in an input vector where all 16 muscles were slightly different (20%) from the training data, and the predicted output vectors suggested that the models were valid. The trained networks were then used to perform two separate simulations: 30% reduction in soleus activity; and removal of rectus femoris. Net 2, in which electromyography was mapped onto joint moments, provided the most reasonable results, suggesting that neural networks can provide a successful platform for both biomechanical modeling and simulation. We believe that this paper has demonstrated the potential of artificial neural networks, and that further efforts should be directed towards the development of larger training sets based on normal and pathological data. PMID- 8429054 TI - Young's modulus of trabecular and cortical bone material: ultrasonic and microtensile measurements. AB - An ultrasonic technique and microtensile testing were used to determine the Young's modulus of individual trabeculae and micro-specimens of cortical bone cut to similar size as individual trabeculae. The average trabecular Young's modulus measured ultrasonically and mechanically was 14.8 GPa (S.D. 1.4) and 10.4 (S.D. 3.5) and the average Young's modulus of microspecimens of cortical bone measured ultrasonically and mechanically was 20.7 GPa (S.D. 1.9) and 18.6 GPa (S.D. 3.5). With either testing technique the mean trabecular Young's modulus was found to be significantly less than that of cortical bone (p < 0.0001). However, the specimens were dried before microtensile testing so Young's modulus values may have been greater than those of trabeculae in vivo. Using Young's modulus measurements obtained from 450 cubes of cancellous bone and 256 cubes of cortical bone, Wolff's hypothesis that cortical bone is simply dense cancellous bone was tested. A multiple regression analysis that controlled for group membership showed that Young's modulus of cortical bone cannot be extrapolated from the Young's modulus vs density relationship for cancellous bone, yet the Young's modulus of trabeculae can be predicted by extrapolation from the relationship between Young's modulus vs density of the cancellous bone. These results suggest that when considered mechanically, cortical and trabecular bone are not the same material. PMID- 8429055 TI - Natural preload of aortic valve leaflet components during glutaraldehyde fixation: effects on tissue mechanics. AB - The mechanics of glutaraldehyde-fixed aortic valve leaflets depend largely on the amount of stress present during fixation. Our previous work has suggested that even when the aortic valve is flaccid, the leaflet components are preloaded. We have, therefore, hypothesized that fixing valve leaflets in this naturally preloaded state will affect the function of their components, the fibrosa and the ventricularis. We have compared the elastic response of fibrosa and ventricularis fixed under 'low' and 'zero' tensile and compressive preload by testing 120 of these layers: (i) fresh, (ii) glutaraldehyde-fixed, and (iii) isolated from whole porcine aortic valve leaflets fixed while intact. In both the radial and circumferential directions, the fibrosa from intact-fixed valves was more extensible than the fresh (39.2 vs 29.2% strain to high modulus phase at p < 0.0122, and 12.7 vs 8.1% strain, at p < 0.0003, respectively). The ventricularis from intact-fixed valves, however, was less extensible than when fresh (35.4 vs 63.7% strain, at p < 0.00001 in the radial direction). The fibrosa must have, therefore, been fixed under compression and the ventricularis under tension, when fixed together in the intact aortic valve cusp. The tensile stresses in the intact-fixed ventricularis produced a greater circumferential elastic modulus than in separately fixed tissue (9.62 vs 4.65 MPa, at p < 0.00001), likely through a fibre recruitment process. Compressive stresses in the fibrosa produced a decrease in the elastic modulus both radially and circumferentially (from 3.79 to 2.26 MPa at p < 0.0023, and from 9.55 to 4.65 MPa at p < 0.00001, respectively). Fixing porcine aortic valves at even minimal tensile and compressive preload, such as that which occurs naturally, significantly alters both the extensibility and the elastic modulus of the valve leaflet components. PMID- 8429056 TI - Collapse of diseased arteries with eccentric cross section. AB - The mechanism of atherosclerotic plaque rupture is not fully understood. Mechanical stress may be one of the factors contributing to the instability of the fibrous plaque cap. The existence of a severe stenosis may lower the transmural pressure enough to cause the collapse of arteries leading to high concentrated compressive and tensile stresses. This study presents quantitative estimates of the stresses and deformations in the collapsed thick-walled artery. The results of large deformation finite element calculations identify the locations of the high stress concentrations and their magnitudes which cannot be precisely predicted under a thin-wall assumption. The maximum compressive stress calculated reached 80% of the Young's modulus for fairly small negative transmural pressures. Results are useful to predict likely locations of the plaque cap rupture due to compressive stresses. The tube law of area as a function of transmural pressure showed a large discrepancy from a thin-wall calculation. The buckling pressure calculated for the outer-to-inner wall surface radius ratio of 1.60 was nearly 30% lower than that of the elastic thin-wall buckling theory. An increase in eccentricity further reduced this buckling pressure. The results indicate that a thick plaque which is eccentric increases the likelihood of collapse of stenotic arteries. PMID- 8429057 TI - Effect of initial upper-limb alignment on muscle contributions to isometric strength curves. AB - Male and female isometric strength curves for elbow fixation, shoulder flexion, and wrist supination-pronation are obtained during systematic variation in arm configuration. The shape of a given moment-angle curve is found to be a function of the orientations of joints kinematically coupled to the primary joint. It is also found that female elbow strength curves are shifted toward flexion with respect to male elbow-strength curves, suggesting that the in situ rest length of upper-limb muscles relative to joint angle may be longer for males than for females. Experimental results were contrasted with simulation results obtained using a three-dimensional musculoskeletal model which estimates the relationships between initial joint orientations, muscle tension-length behavior, and joint moments. In most of the cases, simulation results complimented experimental data and provided insights into likely in situ muscle rest lengths and moments arms, especially for the multiarticular biceps brachii muscle. Where inconsistencies exist between simulated and experimental data, subtle biomechanical complexities within the forearm and the shoulder girdle complex are identified that require future investigation. PMID- 8429058 TI - A theoretical basis for interpreting the force applied to the pedal in cycling. AB - This article presents an analytical technique for decomposing the pedal force in cycling into a muscular component due directly to the net intersegmental moments and a nonmuscular component due to gravitational and inertial effects. The decomposition technique uses the Newton-Euler system of dynamic equations for the leg segments to solve for the two components, given the planar segmental kinematics and the intersegmental moments. Applications of the technique to cycling studies of muscle function, pedalling effectiveness, and optimization analyses based on inverse dynamics are discussed. While this article focuses on the pedal force in cycling, the decomposition method can be directly applied to analyze the reaction forces during a general planar movement of the leg when the segmental kinematics and intersegmental moments are specified. This article also demonstrates the significance of the nonmuscular component relative to the muscular component by performing the decomposition of the pedal forces of an example subject who pedalled at three different cadences against a common work load. The key results were that the nonmuscular components increased in magnitude as the cadence increased, whereas the magnitude of the muscular component remained relatively constant over the majority of the crank cycle. Also, even at the slowest pedalling rate of 70 rpm, the magnitude of the nonmuscular component was substantial. PMID- 8429059 TI - Bone ingrowth: an application of the boundary element method to bone remodeling at the implant interface. AB - Surface bone remodeling theory and the boundary element method are employed to investigate the microstructural remodeling of bone at the bone-implant interface. Three situations are considered: remodeling-induced penetration between the screw threads of an implanted screw, penetration of bone tissue into a slot or cavity in an implant, and the interaction of individual trabeculae in the remodeling processes near an implant. For each case the bone ingrowth is determined as a function of the geometry and the applied load. PMID- 8429060 TI - Bone strain sensation via transmembrane potential changes in surface osteoblasts: loading rate and microstructural implications. AB - A model is developed in which osteoblasts can sense the strains applied to a small region of bone through electrical coupling between adjacent cells. The stress-generated potentials within bone are assumed to occur through streaming potentials, and the coupled network of osteocytes is assumed to act in a manner similar to the classical cable model for nerve cells. In a one-dimensional model, the linear poroelastic equations for motion of the fluid are solved analytically for sinusoidally varying imposed strains, and the streaming potentials are predicted from the fluid flow. The changes in the osteocyte and osteoblast transmembrane potential are given by an analytical solution to the governing equations, and the dependence of the transmembrane potential changes (TPC) on position, loading rate, manner of loading (compression versus bending), and on the degree of cellular coupling is discussed. The model correctly predicts the rate dependence of remodelling established by other investigators. The influence of the electrical parameters within the model indicate that further study of the cellular coupling in bone can yield important new information on bone remodelling. PMID- 8429061 TI - Nonlinear separation of forward and backward running waves in elastic conduits. AB - A new method for the separation of forward and backward running waves in elastic conduits, with possible extension to the arterial system, has been developed. The mathematical model is based on the one-dimensional flow equations which allow the treatment of non-periodic or transient pressure and flow pulses. The method is fully nonlinear, i.e. no linearizing assumptions are made. The method includes the effects of convective acceleration and pressure-dependent vessel compliance. A first approximation for the fluid friction at the wall is also included. The application of the method requires the knowledge of the elastic properties, the instantaneous pressure and flow, as well as the instantaneous spatial derivatives of pressure and flow. Analysis of simulated data shows good results and suggests that the proposed method, unlike previous quasi-nonlinear and frequency domain methods, can be applied to strongly nonlinear and/or nonperiodic flows. The method predicts that if a linear analysis is applied to a nonlinear system errors arise. PMID- 8429062 TI - A method for obtaining repeatable measurements of the tensile properties of skin at low strain. AB - Initial attempts at measuring the low strain tensile properties of skin led to results which were not repeatable. It appeared that strains induced in the sample due to the handling needed to mount it in a simple clamp led to the low precision of the test. A new method which minimized the manipulation needed to mount a sample in a testing machine was then developed. Using this method the skin was aligned with a set of grips in a saline bath. When aligned, the saline was removed and the sample clamped in the grips without the need for additional handling. This approach produced a marked improvement in the repeatability of the tensile properties of skin at low strains. PMID- 8429063 TI - Biomechanics and Galileo Galilei. PMID- 8429064 TI - Symptoms are the body's way of communicating with itself. PMID- 8429065 TI - Symptoms: evidence and experience. PMID- 8429066 TI - A holistic approach to symptom assessment and intervention. PMID- 8429067 TI - Cultural meaning of symptoms. PMID- 8429068 TI - Using a nursing model to manage symptoms: anxiety and the Roy adaptation model. PMID- 8429069 TI - A phenomenological study of couples' infertility: gender influence. PMID- 8429071 TI - Nausea: the forgotten symptom. PMID- 8429070 TI - Therapies for dyspnea relief. PMID- 8429072 TI - Diarrhea: a symptom. PMID- 8429073 TI - Structural characterization of polypeptides and proteins by combination of capillary electrophoresis and (252)Cf plasma desorption mass spectrometry. AB - An efficient and sensitive method for the isolation and transfer of peptides and proteins from capillary zone electrophoresis separation for subsequent analysis by 252Cf plasma desorption mass spectrometry was developed. Sample isolation on to nitrocellulose-coated targets for mass spectrometric analysis is performed by using a stainless-steel microtube pre-filled with aqueous buffer solution, to which the capillary end is connected, and the peptide is collected by applying a suitable transfer voltage according to the separation voltage. Low-and sub picomolar sample amounts were isolated with high transfer efficiency and reproducibility, without the necessity for independent determination of electroosmotic flow-rates. Plasma desorption mass spectra of several peptides and proteins showed predominantly intact molecular ions; however, for several peptides partial oxidative modification was found which can be accounted for by the electrophoretic separation and/or transfer conditions. First applications to peptides and proteins show the feasibility of this off-line combination for primary structure characterization, such as by in situ chemical modification and enzymatic proteolysis reactions on the sample target prior to mass spectrometric analysis. PMID- 8429074 TI - Borate complexation of flavonoid-O-glycosides in capillary electrophoresis. II. Separation of flavonoid-3-O-glycosides differing in their sugar moiety. AB - Capillary electrophoresis was found to give significantly higher efficiency, selectivity and speed than high-performance liquid chromatography for the separation of a mixture of flavonoid-3-O-glycosides differing in their sugar moiety. Boric acid running buffer (0.2 M, pH 10.5) was used for this electrophoretic separation. The migration order in free solution capillary electrophoresis (CE) and the selectivity of these flavonoid-3-O-glycosides can be mainly explained by in situ borate complexation of both the sugar moiety and the cis-1,2-hydroxyl groups on the flavonoid skeleton and, to a lesser extent, by the ionization of hydroxyl groups on the flavonoid skeleton due to alkaline pH conditions. The correlation of the electrophoretic mobilities with the configuration and conformation of the compounds is discussed. PMID- 8429075 TI - Analysis of alkylpyrrole autoxidation products by high-performance liquid chromatography with thermospray mass spectrometry and UV photodiode-array detection. AB - A method employing high-performance liquid chromatography with thermospray mass spectrometry (TSP-MS) and photodiode-array detection was developed and applied to the analysis of autoxidation products of 2,5-dimethyl-N-alkylpyrroles in aqueous solution under air or 18O2. Numerous oxidation products were separated, characterized and categorized, primarily as (1) non-polar oligomers without incorporated oxygen, and (2) polar, oxygen-containing monomers. Kinetic studies showed that oligomerization was the dominant autoxidation pathway, with production of unstable dimers and trimers and, ultimately, a high-molecular-mass sediment. TSP-MS together with UV and proton nuclear magnetic resonance spectral data revealed that both the dimer and trimer contained a novel methylene bridge. These results suggest that this method is suitable for the analysis of alkylpyrrole autoxidation products that may be relevant to hexane neuropathy and products that are responsible for the instability of fuels in storage. PMID- 8429076 TI - High-performance liquid chromatography versus solid-phase extraction for post derivatization cleanup prior to gas chromatography-electron-capture negative-ion mass spectrometry of N1,N3-bis-(pentafluorobenzyl)-N7- (2 [pentafluorobenzyloxy]ethyl)xanthine, a product derived from an ethylene oxide DNA adduct. AB - N7-(2-Hydroxyethyl)xanthine (N7-HEX), as a standard, has been measured at the low picomole level by the following sequence of steps: (1) derivatization with pentafluorobenzyl bromide; (2) post-derivatization sample cleanup by reversed phase high-performance liquid chromatography (HPLC) or silica solid-phase extraction; and (3) separation/detection by gas chromatography electron-capture negative-ion mass spectrometry (GC-ECNI-MS). The average yield of product from the two sample cleanup procedures applied to 95 pg (0.48 pmol) of N7-HEX was comparable: 60% for HPLC; 56% for solid-phase extraction. The reaction blanks (0 pg N7-HEX) showed an interfering GC-ECNI-MS peak after HPLC cleanup. This problem was not encountered with solid-phase extraction, which, along with its greater convenience, made it the preferred technique for post-derivatization sample cleanup. PMID- 8429077 TI - Application of solid-phase extraction in the determination of U-82217 in rat serum, urine and brain. AB - The techniques of solid-phase extraction (SPE) were applied in the analytical method development for the determination of U-82217, 3-(5-cyclopropyl-1,2,4 oxadiazol-3-yl)-5-[(4-methoxyphenyl)methyl] -imidazo[1,5-a]quinoxalin-4(5H)-one, in rat serum, urine and brain. Samples of serum, urine or brain homogenate containing U-82217 were loaded on C18 SPE columns and eluted with acetonitrile (300 microliters). The prepared samples were analyzed by reversed-phase HPLC using an ODS column with a mobile phase of acetonitrile-water (45:55, v/v) containing 0.12% of acetic acid (pH 6.0 +/- 0.1). The UV absorbance of the column effluent was monitored at a wavelength of 318 nm. The absolute extraction recovery from serum, urine and brain samples was ca. 90%. Linear calibration graphs were obtained over the ranges 5 ng/ml-20 micrograms/ml (serum), 20 ng/ml 20 micrograms/ml (urine) and 50 ng/g-200 micrograms/g (brain). The intra- and inter-assay precision and accuracy were all found to be < 13% at the concentrations evaluated. The strategy in SPE development and the application of this method to the determination of U-82217 in rat serum and brain for a pharmacokinetic study are also discussed. PMID- 8429078 TI - High-performance liquid chromatographic method for the determination of dimethindene in urine. AB - An automated high-performance liquid chromatographic assay, using on-line solid phase extraction, is described for the determination of dimethindene in urine. The solid-phase extraction of the sample (1000 microliters) and the elution of the drug on to the analytical column are performed automatically and concomitantly. The limit of quantification is 5 pmol/ml. PMID- 8429079 TI - Stability of immunoadsorbents comprising antibody fragments. Comparison of Fv fragments and single-chain Fv fragments. AB - Immunoadsorbents comprising Fv fragments specific for hen egg lysozyme were used to recover the enzyme from a 20-fold excess of bovine albumin. We designed automatic equipment to run this model purification system for 100 cycles non-stop and monitored the deterioration of the immunoadsorbents during the cycling procedure. Only minor losses (approximately 25%) in the immunoadsorbents' capacity were detected; this correlated well with ligand loss (measured by enzyme linked immunosorbent assay) which was approximately 0.2% per cycle. A surprising finding was that the use of "single-chain" Fv fragments conferred only a minor advantage with respect to stability of the immunoadsorbents. PMID- 8429080 TI - High-performance liquid chromatographic enantioseparation of glycyl di- and tripeptides on native cyclodextrin bonded phases. Mechanistic considerations. AB - Di- and tripeptides containing at least one glycine moiety were separated into enantiomers on alpha-, beta- and gamma-cyclodextrin bonded phases after their precolumn derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl). It is shown that the choice of a suitable cyclodextrin bonded phase used with a nonaqueous polar mobile phase offers a wide range of possibilities to optimize enantioselectivity. Use of the FMOC derivative greatly enhances sensitivity, stability and enantioselectivity. It is postulated that under reversed-phase conditions the inclusion complex formation between the hydrophobic aromatic part of the FMOC-functionalized peptides and non-polar cyclodextrin cavity interior is the major factor contributing to retention. However, under these conditions there seems to be insufficient interaction between the hydrophilic peptide chain and cyclodextrin hydroxyls. Hence no chiral recognition and enantiomeric separation is observed. In systems operating with polar organic mobile phases the inclusion complex is suppressed as the cyclodextrin cavity is largely occupied by the mobile phase. The enantioselectivity observed is caused by hydrogen bonding between peptide chain and the hydroxyl groups at the mouth of the cyclodextrin. The stereospecific interactions depend very strongly on the cyclodextrin size, the length of the peptide chain and the mobile phase composition. PMID- 8429081 TI - Chemometrics in bioanalytical sample preparation. A fractionated combined mixture and factorial design for the modelling of the recovery of five tricyclic amines from plasma after liquid-liquid extraction prior to high-performance liquid chromatography. AB - A general systematic approach is described for the chemometric modelling of liquid-liquid extraction data of drugs from biological fluids. Extraction solvents were selected from Snyder's solvent selectivity triangle: methyl tert. butyl ether, methylene chloride and chloroform. The composition of a mixture of the three extraction solvents was varied and the extraction yield (recovery) of a group of tricyclic amines was measured at all compositions selected. Two process variables, the extraction time and the extraction intensity, were varied simultaneously with the mixture variables to study their influence and their interaction with the mixture composition. The combined mixture and factorial design statistical techniques obtained in this way enabled the recovery to be modelled as a function of both the composition of the extraction liquid and the process variables. The models were assessed with regard to both descriptive and predictive capacities. The results showed that structurally related compounds may demonstrate different partitioning behaviour with regard to both mixture variables and process variables. It was concluded that mixtures of solvents result in higher extraction efficiencies for the amines. A positive effect on the extraction efficiency was demonstrated by the extraction intensity process variable and extraction time. A positive effect on the extraction efficiency was demonstrated by an interaction between extraction intensity and time. Mixture models in which process variables were introduced were recognized as being very suitable for modelling liquid-liquid extraction systems. PMID- 8429082 TI - Novel affinity separations based on perfluorocarbon emulsions. Development of a perfluorocarbon emulsion reactor for continuous affinity separations and its application in the purification of human serum albumin from blood plasma. AB - Perfluorocarbon affinity emulsions are generated by the homogenisation of a perfluorocarbon oil with a polymeric fluorosurfactant previously derivatised with an affinity ligand and subsequently cross-linked in situ. This procedure gives rise to a novel liquid affinity adsorbent that can be used for continuous protein purification. Discrete emulsion droplets were found to be unstable when pumped for prolonged periods; however, when flocculated, the emulsion floccules with diameters of around 125 microns, were very stable and sedimented faster. A four stage reactor unit (perfluorocarbon emulsion reactor for continuous affinity separations, PERCAS) was designed and constructed to carry out continuous separations, and exploited the unusual properties of the absorbent, i.e. liquid nature and high density. Each of the four stages of PERCAS consisted of a mixing tank, for contacting between emulsion phase and aqueous phase, adjacent to a settling tank for the subsequent separation of emulsion from the aqueous phase. Using PERCAS adsorption, washing, elution and re-equilibration of the emulsion could be carried out continuously with emulsion recycle. Using single-component adsorption of human serum albumin to a perfluorocarbon affinity emulsion derivatised with the triazine dye C.I. Reactive Blue 2, PERCAS was optimised with respect to flow-rates and input concentrations. The work was then extended to the continuous purification of essentially homogeneous human serum albumin from blood plasma. PMID- 8429083 TI - Enantiospecific drug analysis via the ortho-phthalaldehyde/homochiral thiol derivatization method. AB - Pre-column derivatization with o-phthaldialdehyde and an optically active thiol has hitherto been used mainly for liquid-chromatographic chiral separation of amino acids. Chiral separation of non-amino-acid primary amines, especially of pharmaceuticals, via this approach has been largely ignored. We have therefore examined the applicability of the method to the chiral resolution of several pharmaceutical amines. o-Phthaldialdehyde and four commercially available homochiral thiols were used to study the separation of the enantiomers of amphetamine, p-hydroxyamphetamine, p-chloroamphetamine, 3-amino- 1-phenylbutane, 3-amino-1-(4-hydroxyphenyl)-butane, mexiletine, tocainide, tranylcypromine and rimantadine. The resulting highly fluorescent isoindole derivatives were resolved on a Waters Nova-Pak C18 column using mobile phases consisting of mixtures of methanol, a sodium acetate buffer and acetonitrile, and the column effluent was monitored using fluorescence or UV detection. In some cases the fluorescence and/or the UV absorbance of the two diastereomers were unequal. It was found that the resolution of most of the amines could be optimized by varying the homochiral thiol in the derivatization step. This method of chiral separation may have wide applicability in enantiospecific drug analysis of non-amino-acid primary amines due to its simplicity and the high sensitivity it provides. PMID- 8429084 TI - Multimycotoxin detection and clean-up method for aflatoxins, ochratoxin and zearalenone in animal feed ingredients using high-performance liquid chromatography and gel permeation chromatography. AB - A sensitive and reliable method is described for the determination of aflatoxins B1, B2, G1 and G2, ochratoxin A and zearalenone in animal feed ingredients. A multi-toxin extraction and clean-up procedure is used, with dichloromethane-1 M hydrochloric acid (10:1) being used for the extraction and gel permeation chromatography being used for the clean-up. The liquid chromatographic method developed for the separation of the six mycotoxins involves gradient elution with a reversed-phase C18 column and fluorescence detection. Recoveries, repeatability and reproducibility have been determined on maize, palm and wheat. The detection limits varied depending on the type of feed. PMID- 8429085 TI - Chemical characterization of cellulose acetate by non-exclusion liquid chromatography. AB - The physical properties of cellulose acetate polymers are not governed solely by their average chemical properties, but are also dependent upon molecular mass and acetyl distributions. Currently, most analytical techniques that are routinely used to characterize cellulose esters, provide only a mean value of the property that is measured. Non-exclusion liquid chromatography was investigated as a means of analyzing cellulose acetate polymers over a compositional range from 37-43% (w/w) acetyl. Under optimum conditions this technique was highly selective for the chemical composition of these polymers, providing information on both mean acetyl composition and acetyl distribution. The purpose of this paper is to describe chromatographic variables required to successfully characterize cellulose acetate. PMID- 8429086 TI - High-performance liquid chromatography-thermospray mass spectrometry of ten sulfonamide antibiotics. Analysis in milk at the ppb level. AB - Ten sulfonamide antibiotics including sulfanilamide (SNL), sulfamethazine (SMZ), sulfamethizole (SMTZ), sulfachloropyridazine and sulfaquinoxaline (SQX), were analyzed by thermospray (TSP) mass spectrometry on-line with a high-performance liquid chromatography-UV detection system. Except for the pairs SMZ-SMTZ and sulfadimethoxine-SQX, the standards were resolved in both the UV and TSP profiles. Co-eluting compounds could be differentiated in TSP by their different relative molecular masses. The [M+H]+ ion was the base peak for all the standards except SNL, which showed an [M+NH4]+ ion. Collision-induced dissociation of the [M+H]+ ions afforded daughter ion spectra characterized by common ions at m/z 92, 108 and 156, and ions derived from the amine substituent ([MH-155]+). TSP detection limits [signal-to-noise ratio (S/N) > 3] were below 20 ng (scan mode), 2 ng (selected reaction monitoring, daughter ions from [M+H]+) and 400 pg (selected ion monitoring). UV detection limits were ca. 2 ng (S/N > 5). Results obtained from the multi-residue analysis of spiked cow milk samples at the low ng/ml level are presented. PMID- 8429087 TI - Gas chromatographic determination of trace nitrite after derivatization with ethyl 3-oxobutanoate. AB - A gas chromatographic method was developed for the determination of nitrite using ethyl 3-oxobutanoate as a derivatization reagent. Nitrite was reacted with ethyl 3-oxobutanoate in the presence of hydrochloric acid to form ethyl 2-hydroxyimino 3-oxobutanoate (EHOB) quantitatively. The resulting EHOB was extracted with ethyl acetate and then determined sensitively by gas chromatography with electron capture detection. This method has been applied successfully to the determination of nitrite in river water and human saliva, with a detection limit of 2 ng/ml and recoveries of 94-100%. PMID- 8429088 TI - [Medical education and research in the era of internationalization]. PMID- 8429089 TI - [The regulation of glycogen synthesis in the fetal rat liver in the near term]. AB - In order to clarify the mechanism of regulation of glycogen metabolism in fetal rat liver during gestation, we measured glycogen synthetase activity, and the binding capacity of the insulin receptor. We also examined the possibility of glycogen synthesis through gluconeogenesis. Major findings are as follows: 1) Glycogen content in fetal liver increased dramatically during gestation and peaked on Day 21 of gestation (one day before delivery). 2) Both the total glycogen synthetase activity (a: active enzyme+b: inactive enzyme) and the ratio of the activities of a to a+b also rose during gestation, suggesting that an increase in the active form of glycogen synthetase is a major cause of the increase in the amount of glycogen in fetal liver. 3) Insulin is known to stimulate glycogen synthesis by activating glycogen synthetase. However, the number and the affinity constant of the insulin receptor in fetal liver did not changed, although the concentration of serum insulin increased during gestation. 4) The ratio of glycogen synthesized from serum glycerol relative to that from serum glucose plus glycerol increased from 8% on Day 19 to 22% on Day 21. Since stimulation of gluconeogenesis is considered to result in an accumulation of glucose-6-phosphate, a potent activator for glycogen synthetase, glyconeogenesis from glycerol may play an important role in glycogen storage in fetal liver. PMID- 8429090 TI - [A study of brain temperature in patients with severe head injuries]. AB - The significance of brain temperature was investigated in patients with severe head injuries. In thirty patients in a state of coma due to severe head injury (Glasgow coma scale of less than 8), brain temperature (Tb), rectal temperature (Tr), and intracranial pressure (ICP) were measured. In 21 out of these patients, cerebral blood flow (CBF) was measured by means of transcranial Doppler effect (TCD), and pulsatility index (PI) was calculated from TCD findings. The outcome was assessed by the Glasgow outcome scale (GOS) 1 month after the head injury. The patients were classified into 4 groups according to the differences (delta T) between Tb and Tr. Group I (11 patients) was defined as always having a higher Tb than Tr, Group II (14) as always having a higher Tr than Tb, Group III (3) was as having no significant differences between Tb and Tr, and Group IV (2) was as having delta T change from plus to minus or from minus to plus during the course of monitoring. As regards the intracranial lesions, no significant differences among these groups were recognized. P.I, at 6 hours after operation, was 1.1 +/- 0.1 (N = 10) in Group I, and was 3.4 +/- 1.9 (N = 9) in Group II. The difference between these two values was statistically significant. The outcome of Group I was better than that of the other groups. The brain temperature is thought to reflect the CBF in the human brain.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429091 TI - [Alveolar epithelial permeability in bronchial asthma in children. An evaluation by 99mTc-DTPA inhalation scintigraphy]. AB - To evaluate alveolar epithelial permeability (kep) in children with bronchial asthma, 99mTc-DTPA (diethylene triamine penta acetate) aerosol lung inhalation scintigraphies were performed. There was no correlation between the kep value and the severity of asthma. On the other hand, out of 10 the cases which had no aerosol deposition defect in the lung field, 4 showed high kep values on the whole lung field and 7 had high kep value areas, particularly apparent in the upper lung field. These results suggest that even when the central airway lesions are mild, severe damage exists in the alveolar region of the peripheral airway. PMID- 8429092 TI - Dermatomyositis-polymyositis and malignancy. Is there a direct relation? AB - An association between dermatomyositis-polymyositis and malignancy has been reported. However, controversy exists regarding a direct causal relationship between the two diseases. We describe a patient who had the concurrent onset and parallel clinical course of dermatomyositis and malignancy, and discuss the possible mechanisms for the association of the two disorders. PMID- 8429093 TI - [The segmental approach by two-dimensional echocardiography to diagnosis in congenital heart disease]. PMID- 8429094 TI - [Recent advances in the diagnosis of arrhythmias. Signal-averaged electrocardiography and its applications]. PMID- 8429095 TI - [Accuracy in computer diagnosis of electrocardiogram]. PMID- 8429096 TI - Sonic stress as one of the cardinal risk factors for atherosclerosis. PMID- 8429097 TI - Detection of tumor necrosis factor-alpha-positive cells in cerebrospinal fluid of patients with HTLV-I-associated myelopathy. AB - Tumor necrosis factor (TNF)-alpha-positive cells constituted 1.6-18% and 8.2 23.5% of the total number of cerebrospinal fluid cells from six of 12 patients with HTLV-I-associated myelopathy and in all samples obtained from inflammatory cases, respectively. However, in non-inflammatory cases no TNF-alpha-positive cells were detected. These results suggest that some of the infiltrating CSF cells produce TNF-alpha, which plays a role in host immune defenses against causative agents including HTLV-I and in lesion formation within the central nervous system in inflammatory diseases. PMID- 8429098 TI - The influence of interleukin-2 on vasopressin and oxytocin gene expression in the rodent hypothalamus. AB - There is growing evidence of interactions between the central nervous system and the immune system. We present evidence that the cytokine interleukin-2 (IL-2) influences expression of the genes encoding the neuropeptides vasopressin (VP) and oxytocin (OT) in the hypothalamus of the nude mouse. A single injection of recombinant mouse IL-2 (rmIL-2) caused a significant increase in VP and OT mRNA levels in the hypothalamus of nude mice. This effect was specific to the nude mouse. These observations stress the potential value of the nude mouse for studying interactions between the central nervous system (CNS) and the immune system. PMID- 8429099 TI - Neuroenteric peptides affect natural killer activity by intestinal lamina propria mononuclear cells. AB - The effect of the neuropeptides bombesin and vasoactive intestinal peptide, as well as neurotensin, on natural killer activity by lamina propria mononuclear cells isolated from histologically normal mucosa was assessed. Bombesin and vasoactive intestinal peptide were found to dose-dependently stimulate natural killer activity against Caco-2 colon carcinoma target cells, especially in a short incubation assay, whereas no alterations of cytotoxicity were found against K-562 target cells. Neurotensin, on the contrary, was not found to affect this type of cellular cytotoxicity. Differences in the modulatory effects of these gastrointestinal peptides on natural killer activity by lamina propria mononuclear cells may be related to the distinct localization differences of these peptides within the bowel. The present findings illustrate the existence of neuro-immune interactions at the intestinal mucosa level. PMID- 8429100 TI - Fluctuation of HTLV-I proviral DNA in peripheral blood mononuclear cells of HTLV I-associated myelopathy. AB - To assess the immunopathological significance of the increased replication of human T-cell leukemia virus type I (HTLV-I) in HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) we investigated the dynamics of HTLV-I proviral DNA in peripheral blood mononuclear cells (PBMC) of HAM/TSP patients at different clinical stages. We compared the dynamics to those of asymptomatic HTLV-I carriers (AC). The estimation of the amount of HTLV-I proviral DNA was carried out by quantitative polymerase chain reaction of serially diluted DNA samples where it was feasible to titrate 0.04-80 copies per 100 PBMC. The proviral DNA quantified in six patients with HAM/TSP was 2-20 copies per 100 PBMC, while that in eight cases of AC was 0.04-8 copies per 100 PBMC. Thus, the amount of HTLV-I proviral DNA in HAM/TSP patients was 3-50 times as high as that of AC. When we followed up HAM/TSP patients for 1-3 years, the amount of HTLV-I proviral DNA fluctuated from 4 to 10-fold. These data suggest that the rate of HTLV-I replication increases in HAM/TSP and the amount of HTLV-I proviral DNA fluctuates in their clinical course. Fluctuation in the amount of HTLV-I proviral DNA may reflect dynamics of HTLV-I infected cell proliferation and immunological suppression in vivo in HAM/TSP patients. PMID- 8429101 TI - Anti-lymphocyte antibodies and circulating immune complexes in the sera of patients with myelopathy associated with human T lymphotropic virus type-I. AB - We measured levels of circulating immune complexes in the human T-lymphotropic virus type I (HTLV-I)-associated myelopathy (HAM) by Raji cell assay and C1q binding assay using an enzyme-linked immunosorbent assay (ELISA). The levels of anti-lymphocyte antibody were also measured using normal donor peripheral blood T lymphocytes. The levels of anti-lymphocyte antibodies were significantly higher in the sera of patients with HAM compared to controls (P < 0.01). The concentrations of immune complexes measured by Raji cell assay were also significantly higher in the HAM patients' sera than in controls. However, when levels of immune complexes were measured by C1q binding assay, there was no significant difference between HAM patients and controls. There was no significant difference in the levels of anti-lymphocyte antibodies and immune complexes between HTLV-I carriers and controls. Circulating immune complexes detected by the Raji cell assay did not include HTLV-I p-19 as detected by the indirect immunofluorescent method. Levels of anti-lymphocyte antibody were correlated with levels of circulating immune complexes as detected by the Raji cell assay in the sera of patients with HAM. These findings indicate that anti lymphocyte antibodies and circulating immune complexes are present in the sera of HAM patients, and that the levels of complexes detected by Raji cell assay may reflect anti-lymphocyte antibody levels. PMID- 8429102 TI - Stress-induced modulation of the primary cellular immune response to herpes simplex virus infection is mediated by both adrenal-dependent and independent mechanisms. AB - A murine model of herpes simplex virus (HSV) infection was used to examine the role of the adrenal gland in restraint stress-induced suppression of viral immunity. Adrenal-dependent mechanisms were important for suppressing the generation of HSV-specific cytotoxic T lymphocytes (CTL) but not the associated diminished lymphadenopathy in response to local HSV infection. While exogenous corticosterone administration alone was unable to suppress lymphadenopathy and CTL generation in adrenalectomized mice, an adrenal-independent mechanism induced by restraint stress functioned in synergy with corticosterone to suppress lymphadenopathy and CTL development. These results suggest that both adrenal dependent and independent mechanisms contribute to stress-induced modulation of HSV immunity. PMID- 8429103 TI - Increase in IL-6, IL-1 and TNF levels in rat brain following traumatic lesion. Influence of pre- and post-traumatic treatment with Ro5 4864, a peripheral-type (p site) benzodiazepine ligand. AB - The effects of fluid percussion trauma on brain interleukin (IL)-6, IL-1 and tumor necrosis factor-alpha (TNF-alpha) levels have been studied. In the cortex and hippocampus of control and sham-operated rats, the levels of these cytokines were very low (below 4 units/mg protein) and constant. IL-6 and IL-1 levels in the ipsilateral cortex increased rapidly following trauma to reach a maximum of 350 and 16 units/mg protein, respectively, 8 h after the lesion, remained elevated until 18 h and decreased thereafter to basal values. TNF-alpha levels were maximally elevated (12 units/mg protein) at 3 h and 8 h and returned to basal values by 18 h. Qualitatively similar changes, but with 25-80-fold smaller amplitude, were seen in the contralateral cortex and in the ipsi- and contralateral hippocampus. The levels of IL-6 in the plasma of sham-operated and lesioned rats were only slightly elevated, whereas IL-1 and TNF-alpha were undetectable. Histological studies of brain tissue at early stages after trauma demonstrated an acute hemorrhage associated with neutrophil invasion. The administration of Ro5 4864 (0.5 mg/kg i.p.), a specific ligand of p (peripheral type benzodiazepine) binding sites, did not result in any significant effect on the levels of IL-6, IL-1 or TNF-alpha in the brain of control or sham-operated animals. However, when administered 24 h before or 15 min after trauma, this benzodiazepine enhanced the increase of these cytokines by 2-4-fold in the ipsilateral cortex.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429104 TI - Modulation of non-specific immunity by hippocampal stimulation. AB - Rats subjected to electrical stimulation of hippocampus (30 min/day for 4 days) showed an increase in the neutrophils in the peripheral blood when compared to sham (P < 0.01) and controls (P < 0.001). They also showed a significant decrease in lymphocytes, when compared to control rats (P < 0.001). Both sham and stimulated animals showed a significant decrease in total white blood cell count when compared to controls (P < 0.001). The phagocytic index of stimulated animals showed a significant increase from control (P < 0.001) and sham (P < 0.001). In addition, the stimulated animals showed a significant (P < 0.001) decrease in plasma corticosterone level when compared to sham and controls. PMID- 8429105 TI - Human muscle acetylcholine receptor reactive T and B lymphocytes in the peripheral blood of patients with myasthenia gravis. AB - The prevalence of T and B cells reactive with the acetylcholine receptor (AChR) of human skeletal muscle was studied in 33 patients with myasthenia gravis (MG), 18 patients with other neurological diseases (OND) or autoimmune disorders (AD) and 27 age- and sex-matched healthy controls. T cell stimulation was estimated by enumerating cells secreting interferon (IFN)-gamma and interleukin (IL)-2 in response to the AChR, whereas B cell reactivity was estimated by enumerating cells secreting IgG antibodies binding to the AChR. AChR-reactive T cells were increased in the peripheral blood of patients with MG as compared to patients with OND, AD and healthy individuals. Of the patients with MG, 29/33 (87.7%) had numbers of IFN-gamma secreting cells higher than the mean +/- 2 SD of the mean of controls as compared to 4/18 (22.2%) of patients with OND or AD and 2/27 (7.4%) of the controls. The mean value of the numbers of AChR-reactive T cells in the patients with MG was 19.6/10(5) PBMC, corresponding to 1/5100 PBMC. Comparable results were obtained also for IL-2-secreting cells. Anti-AChR IgG antibody secreting cells were detected in the blood of 30/33 (91%) of the patients with MG, 3/18 (16.7%) of the patients with OND or AD and 2/25 (8%) of the controls. The mean value of the antibody-secreting cells in MG was 11.7 cells/10(6) PBMC corresponding to 1/70,400 PBMC in the patients with MG, compared to a mean value of antibody-secreting cells in the patients with OND or AD of 0.33 and controls of 0.16 cells/10(6) PBMC. PMID- 8429106 TI - Cerebrospinal fluid interferon-gamma is increased in HTLV-I-associated myelopathy. AB - Serum and cerebrospinal fluid (CSF) levels of interferon-gamma (IFN-gamma) were assayed with an immunoenzymetric method in 16 patients with HTLV-I-associated myelopathy (HAM). IFN-gamma levels were elevated in serum of two patients and in CSF of nine. Although there was no significant association between the clinical features and the CSF IFN-gamma levels, the mean levels of CSF IgG and CSF anti HTLV-I antibody titer were higher in patients with elevated CSF IFN-gamma levels than in those with normal CSF IFN-gamma levels. PMID- 8429107 TI - Regulating continuing medical education. PMID- 8429108 TI - The drug regulatory and review process in Guyana. AB - After the old "Sale of Food and Drugs" Ordinance, Cap. 144 was repealed, the new Food and Drugs Act was enacted in 1971. This new Act has considerable flexibility and gives the Minister extensive authority to make Regulations (for carrying out the purposes and provisions of the Act). The Act controls the manufacture, importation, sale, advertising, labeling, packaging, and distribution of drug samples, and the testing of drugs. The Act also controls raw materials and finished products of drugs at the point of entry into the country, with a single agency coordinating both the inspection and analytical services. Developing countries could ensure the procurement of safe, good quality, and effective drugs and devices with the enactment of a similar Food and Drugs Act only. Rapid assessment of Drug Safety, Quality and Efficacy is done through Guyana's participation in the WHO Certification Scheme on the Quality of Pharmaceutical Products moving in International Commerce. This certification scheme is highly commendable especially to third-world countries. The Food and Drug Regulations (1977) have several unique features for drug, cosmetic and device control and they allow for a system of centralized control with limited staff to enforce the legislation. In summary, enforcement of legislative control of imported pharmaceuticals and product evaluation can be considered strong points in the drug regulatory and review process in Guyana. A cautious attitude is observed so as to ensure efficacy, safety, and quality of drugs entering the market. This Drug Regulatory and Review Process is recommended for implementation by third world countries with outdated drug legislation.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429109 TI - A legal curriculum for the clinical pharmacologist: II. The court system and grants and contracts. PMID- 8429110 TI - The pharmacokinetics of isoproterenol in critically ill pediatric patients. AB - The pharmacokinetics of isoproterenol (ISO) in infants and children have never been reported. The authors studied ISO pharmacokinetics in two disparate groups of pediatric intensive care unit patients: postoperative cardiac patients (POC, n = 10), and reactive airway disease patients (RAD, n = 9). In all, 44 blood samples were taken at steady-state from the 19 patients, whereas from 15 patients samples were also taken just before and after discontinuation of ISO infusion. There were 12 male and 7 female patients in the study, and their ages ranged from 2 days to 14 years. The average ISO dosing rate was 0.30 micrograms/kg/minute for the whole study population, ranging from 0.01 to 5.5 micrograms/kg/minute. The POC patients received a significantly lower dosing rate than the RAD patients (0.029 +/- 0.002 vs 0.50 +/- 0.21 micrograms/kg/minute, P < .0001); the average steady-state plasma concentrations of ISO were also lower in the POC patients (1.3 +/- 0.3 versus 13.9 +/- 4.9 ng/mL, P < .0001). The steady-state plasma concentration, normalized to a dosing rate of .05 micrograms/kg/minute, was 1.9 +/- 0.3 ng/mL for all patients, and the clearance was 42.5 +/- 5.0 mg/kg/minute. Postoperative cardiac patients had a significant higher normalized steady-state plasma concentration and moderately significant lower clearance than did RAD patients (2.1 +/- 0.3 versus 1.7 +/- 0.4 ng/mL, P < .05 and 33.2 +/- 4.9 versus 48.4 +/- 7.3, P < .06, respectively). The average plasma half-life of ISO was 4.2 +/- 1.5 minutes, and the volume of distribution was 216 +/- 57 mg/kg.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429111 TI - Effect of gemfibrozil on fatty acids in lipid fractions of plasma from patients with hypertriglyceridemia. AB - The fatty acid composition of plasma triglycerides, phospholipids, and cholesterol esters have been studied during gemfibrozil (G)-induced lipid lowering treatment in six patients with hypertriglyceridemia. Gemfibrozil caused significant plasma triglyceride reductions from 776 +/- 573 to 226 +/- 82 mg/dL (P < .001). Gemfibrozil therapy also caused significant changes in the plasma lipid fatty acid composition, mainly by increasing palmitoleic acid (16:1) in triglycerides (2.1-5.6%) and cholesterol esters (2.2-3.7%), concomitant with the significant decrease in the content of the linoleic acid (18:2) in phospholipids (20.1-16.0%) and triglycerides (18.0-15.2%). The ratio of unsaturated fatty acids (20:3 + 20:4/18:2) was increased in the phospholipid fraction. These findings support a G effect on the desaturation of fatty acids in patients with hypertriglyceridemia. Because fatty acid composition was significantly altered by G, those biologic systems dependent on the pattern of fatty acids may be modified in clinical important ways. PMID- 8429112 TI - Evidence of a partial escape of renin-angiotensin-aldosterone blockade in patients with acute myocardial infarction treated with ACE inhibitors. AB - Angiotensin-converting enzyme (ACE) inhibitors have been designed to block the renin-angiotensin system and can represent an effective therapeutic approach in those settings where such a system is active, such as myocardial infarction. In a randomized placebo-controlled study, 10 patients with acute myocardial infarction allocated to treatment with increasing doses of zofenopril calcium and 10 patients allocated to placebo were studied in hospital, within 24 hours from symptoms, during 11 sampling periods to assess the time course of ACE inhibition and renin-angiotensin-aldosterone blockade. Zofenopril administration was followed by a dose-dependent inhibition of in vitro ACE activity (7.5 mg, 65%; 15 mg, 89%; 30 mg, 94.5%) and a progressive increase in plasma active renin. Conversely, plasma aldosterone decreased during the first 3 days of treatment and then returned toward baseline values, as did blood pressure, despite a persistent inhibition of ACE. The present data suggest the existence of an interesting dissociation between the time-course of ACE inhibition and that of blockade of the renin-angiotensin system in patients with acute myocardial infarction. This discrepancy could arise from the combination of an only partial in vivo ACE inhibition and the compensatory increase in plasma renin that occurs during treatment with ACE inhibitors. A better understanding of this relationship would seem to be useful in addressing the correct use of ACE inhibitors in patients with acute myocardial injury. PMID- 8429113 TI - Nisoldipine: a new dihydropyridine calcium-channel blocker. AB - Nisoldipine is a new calcium-channel blocker of the dihydropyridine subclass, with a chemical structure similar to nifedipine. It has been used in clinical trials to assess its efficacy and safety in patients with hypertension, angina pectoris, and congestive heart failure. Similar to other dihydropyridines, nisoldipine is a potent peripheral and coronary dilator. The most optimal dosage regimen has not been established in clinical trials. The drug appears to have a favorable side-effect profile. PMID- 8429114 TI - Effects of oral cimetidine or ranitidine on the pharmacokinetics of intravenous enoxacin. AB - Enoxacin is a quinolone antibacterial agent currently being developed for oral and intravenous treatment of bacterial infections. Ten healthy subjects received a single 400-mg intravenous dose of enoxacin alone, with 300 mg (four times daily) oral cimetidine and with 150 mg (twice daily) oral ranitidine. Serial blood and urine samples were collected over a 48-hour period. Plasma and urine enoxacin concentrations were determined using a validated high-performance liquid chromatographic method. Mean enoxacin plasma concentrations were higher after administration of enoxacin with cimetidine than those measured after enoxacin alone or enoxacin with ranitidine. Cimetidine coadministration reduced enoxacin renal clearance by 26% and systemic clearance by 20%, and resulted in a 30% increase in elimination half-life. In contrast, concurrent ranitidine therapy did not significantly alter the pharmacokinetics of intravenous enoxacin. PMID- 8429115 TI - Phase I study of DQ-2556, a new parenteral 3-quaternary ammonium cephalosporin antibiotic. AB - The safety and pharmacokinetic properties of DQ-2556, a new parenteral cephalosporin, were evaluated using healthy volunteers after 5-minute intravenous infusion of doses of 250, 500, 1000, or 2000 mg and a 1-hour infusion of 2000 mg, and an intramuscular dose of 500 mg. The half-lives of DQ-2556 ranged from 1.64 to 2.15 hours. The peak serum concentrations and area under the curve values were linearly correlated to the doses. The mean urinary recoveries were 80.0 to 85.5% of a dose within 24 hours. Salivary concentrations of the drug were low. There was no accumulation of DQ-2556 after 9 administrations every 12 hours. DQ-2556 was well tolerated. PMID- 8429116 TI - Inhibition of gentamicin uptake into cultured mouse proximal tubule epithelial cells by L-lysine. AB - Gentamicin uptake and toxicity was studied in a nontransformed cell line obtained from the S1 segment of the proximal tubule epithelium of a transgenic mouse. Cytotoxicity was assayed using the dye 3-(4,-5dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT). Gentamicin uptake was assayed by a fluorescence polarization assay. No differences in toxicity were found among cells incubated for 4 hours in complete culture medium, enriched Kreb's buffer alone, or enriched Krebs' buffer with added 300 micrograms/mL gentamicin, 0.5 mmol/L L-lysine, or gentamicin plus L-lysine. Uptake of 300 micrograms/mL gentamicin was minimal at zero time and increased as a function of time. Uptake of gentamicin at 4 hours was positively correlated with medium gentamicin concentration. Addition of 0.5 mmol/L L-lysine inhibited uptake of 300 micrograms/mL gentamicin 38.9 +/- 10.2%. No other amino acid, including D-lysine or arginine, significantly changed gentamicin uptake. The authors conclude that gentamicin and L-lysine share a specific uptake mechanism located in the apical membrane of renal proximal tubule cells. PMID- 8429117 TI - Twenty-four-hour control of gastric acidity by twice-daily doses of placebo, nizatidine 150 mg, nizatidine 300 mg, and ranitidine 300 mg. AB - This study was carried out to assess the effects on gastric acidity of placebo twice daily (bid), nizatidine 150 mg bid, nizatidine 300 mg bid, and ranitidine 300 mg bid by means of continuous 24-hour intragastric pH monitoring. Twelve patients with duodenal ulcer in remission were randomized to receive in single blind fashion the above medications on four separate occasions, at least 1 week apart. The three active regimens produced higher pH values (P < .001) and maintained gastric pH above 3.0 units for a longer period (P < .001) than placebo in all time intervals but evening. Nizatidine 150 mg bid caused a lower rise in pH than nizatidine 300 mg bid (P < .01) and ranitidine 300 mg bid (P < .05) during both the daytime and the whole 24 hours. In these time windows also the time spent above 3.0 pH units was significantly shorter for the former regimen than for 300 mg bid of both nizatidine (P < .01) and ranitidine (P < .05). There was no difference between the latter two dosing schedules in terms of both potency and duration of action in all the time intervals considered. It is concluded that twice daily doses of H2 blockers are more effective than placebo in reducing gastric acidity. Three hundred milligrams twice daily of both nizatidine and ranitidine produce a significantly greater and longer-lasting acid suppression than 150 mg bid of nizatidine. Our study also confirms the greater effectiveness of H2 antagonists during nighttime than during day-time. PMID- 8429118 TI - Antipyrine kinetics in patients with primary biliary cirrhosis. AB - Fourteen antimitochrodrial antibody-positive patients (13 women, 1 man) with biopsy-proven primary biliary cirrhosis, aged 40 to 71 years (mean, 57 years) weighing 43 to 102 kg (mean, 63 kg), along with 14 age- and sex-matched healthy controls, received a single 1.0- to 1.2-g dose of intravenous antipyrine. Plasma antipyrine levels were determined during a 12- to 24-hour period. Patients' mean serum chemistry values were: albumin, 3.9 g/dL (range, 3.1-4.4) and total bilirubin, 1.9 mg/dL (range, 0.3-10.9). Seven of the fourteen patients had cirrhosis. Mean kinetic variables for antipyrine in controls and primary biliary cirrhosis patients were: Vd, .54 versus .49 L/kg; half-life, 12.0 versus 15.1 hours (P < .07); clearance, .55 versus .41 mL/min/kg (P < .04). Within the primary biliary cirrhosis group, there was no correlation between total bilirubin and clearance (r = .09), nor did clearance vary significantly among histologic categories. Clearance of antipyrine in primary biliary cirrhosis patients is reduced by an average of 25%, but the clinical prognosticators of serum bilirubin levels and histologic grade do not correlate with or predict the degree of clearance impairment. PMID- 8429119 TI - The safety of ranitidine in elderly versus non-elderly patients. AB - The authors conducted a retrospective review of 21 United States trials of ranitidine in acid peptic diseases and compared the adverse events in elderly (> or = 65 years) and nonelderly (< 65 years) patients. Ranitidine dosages ranged from 150 mg/day to 300 mg twice daily for treatment periods of 4 to 52 weeks. Of the 4041 patients included in this review, 402 elderly and 2188 nonelderly patients received ranitidine and 245 elderly and 1206 nonelderly patients received placebo; 29%, 29%, 32%, and 26% of these patients, respectively, reported some type of adverse event. When only drug-related adverse events (as judged by the investigators under blinded conditions) were evaluated, these percentages dropped to 2%, 2%, and 1% and 2%, respectively. Gastrointestinal adverse events (e.g., nausea and diarrhea) and central nervous system adverse events (e.g., headache and dizziness) were the most common (0.7% and 0.8%, respectively), with comparable incidence rates in the elderly and nonelderly patients. The authors conclude that ranitidine is as safe in elderly patients as it is in nonelderly patients. No difference in the incidence of adverse events was found between older and younger patients who received ranitidine or placebo. PMID- 8429120 TI - Pharmacokinetics and bioavailability of the RRR and all racemic stereoisomers of alpha-tocopherol in humans after single oral administration. AB - The plasma and red blood cell pharmacokinetics and bioavailability of the natural source (RRR, d) and all racemic (all rac, dl) stereoisomers of alpha-tocopherol were studied in 12 men in a double-blind randomized crossover study. Subjects were administered two 400-mg soft-gelatin capsules of either RRR or all rac alpha tocopherol. Plasma alpha-tocopherol concentrations were determined by high performance liquid chromatography at various time intervals for up to 96 hours postadministration. Pharmacokinetic modeling of the data showed that alpha tocopherol was absorbed after a 2 to 4 hour lagtime and maximum plasma concentration occurred from 12 to 14 hours postadministration. There were no significant differences in the Ka, t1/2 a, beta, or t1/2 beta between RRR and all rac. Mean plasma alpha-tocopherol concentrations were greater for RRR than all rac from 10 to 96 hours postadministration and significantly greater at 24 hours (P < .05). The red blood cell alpha-tocopherol concentration from the RRR preparation was significantly greater than from the all rac preparation from 24 to 96 hours postadministration with Cmax for RRR (4.8 micrograms/mL) significantly greater than for all rac (4.0 micrograms/mL, P < .05). The RRR AUC0 96 for both plasma and red blood cells were significantly greater than the all rac AUC0-96 (P < .05) indicating a greater bioavailability of RRR versus all rac alpha-tocopherol. This difference in overall bioavailability was apparently not due to a single pharmacokinetic component. PMID- 8429121 TI - Bioavailability studies of drugs with nonlinear pharmacokinetics: II. Absolute bioavailability of intravenous phenytoin prodrug at therapeutic phenytoin serum concentrations determined by double-stable isotope technique. AB - Measurement of the absolute bioavailability of phenytoin (PHT) derived from test doses of phenytoin prodrug (PPD) at therapeutic PHT serum concentrations is complicated by two problems: 1) the area under the serum concentration versus time curve (AUC) produced by a given size of test dose will vary directly with background PHT serum concentration due to the nonlinear pharmacokinetic properties of PHT; 2) PPD is more water soluble than PHT, making renal excretion of PPD more likely. The authors describe a double-stable isotope method that obviates these two problems. Using only six subjects, the authors were able to demonstrate bioequivalence of PHT derived from intravenous PPD with intravenous PHT by current FDA standards for AUC ratio of test/reference formulation (90% confidence intervals between 0.80 and 1.20; ratio > or = 0.80 in > or = 80% of subjects; statistical power to detect a difference of 0.20 with a probability of 0.80). PMID- 8429122 TI - Functional organization of the ventral lateral geniculate complex of the tree shrew (Tupaia belangeri): I. Nuclear subdivisions and retinal projections. AB - This is the first of two papers describing the organization and connections of the ventral lateral geniculate complex (GLv) in the tree shrew. Using a combination of Nissl, Golgi, histochemical, and immunocytochemical methods, we have identified two major divisions (lateral and medial) of GLv, both of which can be further subdivided. The lateral division contains three subdivisions, external, internal and intergeniculate leaflet. The medial division contains two subdivisions, medio-rostral and medio-caudal. All three lateral subdivisions receive input from the retina, the densest terminations being in the external subdivision and intergeniculate leaflet. These projections originate primarily from small retinal ganglion cells, although a few large retinal ganglion cells also project to GLv by way of collateral branches. Each subdivision of GLv has a distinct cytoarchitectonic and immunocytochemical make-up. In general, the level of immunoreactive endings for glutamic acid decarboxylase (GAD), leuenkephalin (ENK), and choline acetyltransferase (ChAT) parallels the distribution of retinal projections. Thus, all three markers are particularly dense in the external subdivision and the intergeniculate leaflet. Cell bodies immunoreactive for ENK are restricted to the external and intergeniculate leaflet subdivisions. The medial subdivisions stain relatively poorly for GAD, ENK, and ChAT, although each has other cytological features that differentiate them from the lateral subdivisions and the adjacent thalamic reticular nucleus. PMID- 8429123 TI - Location, morphology, and central projections of mesencephalic trigeminal neurons innervating rat masticatory muscles studied by axonal transport of choleragenoid horseradish peroxidase. AB - Retrograde and transganglionic transport of horseradish peroxidase conjugated to the B-fragment of cholera toxin (B-HRP) was used to study the location, morphology, and central projections of mesencephalic trigeminal (Me5) neurons innervating rat masticatory muscles. Labeled Me5 cell bodies were found throughout the Me5 nucleus from a level slightly caudal to the trigeminal motor nucleus to the level of the superior colliculus 5 mm further rostrally. Occasionally, labeled Me5 cells were observed in the anterior medullary velum, in the cerebellum, and in the brainstem contralateral to the B-HRP injection. The vast majority of the labeled Me5 cells were pseudounipolar, but multipolar cells were also found. Extensive central projections from labeled Me5 cells could be seen extending from the nucleus of Darkschewitsch rostrally to the C2 segment caudally. Small but consistent projections from Me5 neurons were observed in nuclear islands among the incoming Me5 root fibers. Trigeminal and hypoglossal motor nuclei received direct projections from Me5 cells, but not the facial motor nucleus. The most prominent Me5 projections appeared in the brainstem reticular formation, including the supratrigeminal nucleus. Smaller projections also extended into the main sensory trigeminal nucleus, trigeminal subnucleus oralis, and the nucleus of the solitary tract. PMID- 8429124 TI - Regional and laminar organization of projections from the presubiculum and parasubiculum to the entorhinal cortex: an anterograde tracing study in the rat. AB - The regional and laminar organization of the projections from the presubiculum and the parasubiculum to the entorhinal cortex was analyzed in the rat with the anterograde tracer Phaseolus vulgaris-leucoagglutinin (PHA-L). The projections from the presubiculum were bilateral and confined to layers III and I of the medial entorhinal area (MEA). Both the ispi- and the contralateral projections showed similar distributions and were almost of equal density. Projections to layer III of the entorhinal cortex arose predominantly from superficial layers of the presubiculum, whereas the fibers that reach layer I of the entorhinal cortex appear to originate preferentially from the deep layers of the presubiculum. These fibers also appeared to innervate weakly layer II of MEA. The parasubiculum distributed projections not only to MEA but also to the lateral entorhinal area (LEA), innervating layer II selectively. The innervation of LEA was quite dense and extensive. Very weak projections from the parasubiculum to the contralateral entorhinal cortex were observed in this study. The position of the terminal plexus in the entorhinal cortex was determined by the point of origin along both the dorsoventral and transverse or proximodistal axes of the presubiculum and parasubiculum. Projections from the presubiculum and parasubiculum entered the entorhinal cortex at the level of the injection, or slightly ventral to it, and the main terminal field was always present ventrally to the injection site. The dorsoventral axis of origin thus corresponded to a similarly oriented axis of termination in the entorhinal cortex. The distribution in relation to the origin along the transverse axis was more complex, and differences between the presubiculum and parasubiculum were present. The proximal presubiculum, i.e., the part closest to the subiculum, projected to the most lateral part of MEA and the central part of the presubiculum sent fibers to the most medial part of MEA. The distal part of the presubiculum, i.e., the part that borders the parasubiculum, projected to the central part of MEA. Projections from the portion of the parasubiculum directly adjacent to the presubiculum, the so-called proximal parasubiculum, reached medial parts of MEA, and those originating in the central part distributed preferentially to lateral parts of MEA and adjacent medial parts of LEA. The distal part of the parasubiculum that borders the entorhinal cortex projected mainly to almost the full mediolateral extent of LEA.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429125 TI - Fine structure of calcitonin gene-related peptide immunoreactive synaptic contacts in the thalamus of the rat. AB - Recent studies have shown a prominent calcitonin gene-related peptide immunoreactive (CGRP-ir) pathway extending from the external medial and external lateral para-brachial nuclei to the area surrounding and including the gustatory nuclei in the thalamus, and the cortex and amygdala. The function of the CGRP-ir pathway is not completely understood, but may be involved with the processing of both nociceptive and gustatory information in the thalamus. The purpose of this study was to characterize the nature of the CGRP-ir synaptic contacts in the gustatory nucleus. Electron microscopic examination of CGRP-ir synaptic contacts revealed two classes of CGRP-ir terminals. One class, which was large, formed asymmetric synaptic contacts on dendritic appendages, had many small, round synaptic vesicles, and heavy patches of reaction product which obscured any underlying organelles. Since similar terminals in unstained tissue contained large numbers of dense-cored vesicles, it was concluded that CGRP-ir was contained predominantly in dense-cored vesicles. A second class of CGRP-ir terminals was smaller and made either asymmetric or symmetric synaptic contacts. Both symmetric and asymmetric small terminals contained small, round synaptic vesicles and fewer patches of dense reaction product. Several of the CGRP-ir terminals making symmetric contacts also contained pleomorphic vesicles. There were very few contacts on cell bodies. There were no contacts on other CGRP-ir elements, somal or dendritic, or on axon terminals. None of the CGRP-ir terminal elements were postsynaptic to unlabeled terminals. Axons containing CGRP-ir were primarily unmyelinated, but a few myelinated axons were also seen. PMID- 8429126 TI - Distribution of noradrenaline-immunoreactivity in the brain of the mormyrid teleost Gnathonemus petersii. AB - The distribution of noradrenaline-immunoreactivity in the brain of the mormyrid fish Gnathonemus petersii was studied in order to evaluate the noradrenergic innervation of a number of specialized mormyrid brain regions, including electrosensory centers and a gigantocerebellum. Noradrenaline-immunoreactive (NAi) neurons occur in the hypothalamic paraventricular organ (PVO), the locus coeruleus, and the caudal rhombencephalon. In the PVO, NAi cerebrospinal fluid (CSF)-contacting neurons are located in the same regions where dopamine- and serotonin-containing CSF contacting neurons occur. The locus coeruleus consists, on each side, of at least 30 rather large NAi neurons with ventrolaterally directed dendrites and dorsolaterally coursing axons. In the caudal rhombencephalon, NAi neurons are located in the transition region between the ventromedial motor zone and the dorsolateral sensory zone. The density of NAi fibers is very high in the efferent tract of the locus coeruleus, the medial forebrain bundle, and two telencephalic, one preoptic, and one rhombencephalic subependymal axonal plexus. A marked NAi innervation is present in the dorsomedial and ventral telencephalon, the preoptic region, periventricular hypothalamic and thalamic regions, the midbrain tectum, cerebellar granular layers, the electrosensory lateral line lobe, the rhombencephalic transition region between the sensory and motor zones, and the area postrema. Other regions are more sparsely innervated by NAi fibers, but regions completely devoid of NAi fibers were not observed. Interestingly, NAi fibers form large club endings in some subdivisions of the precerebellar nucleus lateralis valvulae, and parallel fibers in the cerebellar granular layer. Comparison with the distribution of NAi or dopamine-beta-hydroxylase-immunoreactivity in other species shows that all teleosts studied to date have noradrenergic cells in the locus coeruleus and the caudal rhombencephalon. However, NAi CSF-contacting PVO cells have been described only in the teleost Gnathonemus petersii and the lizard Gekko gecko (Smeets and Steinbusch: J. Comp. Neurol. 285:453-466, '89). It is possible that they might pick up catecholamines as well as serotonin from the CSF, into which monoamines might be released by telencephalic and preoptic subependymal axonal plexuses. PMID- 8429127 TI - Immunocytochemical localization of midbrain estrogen receptor- and progestin receptor-containing cells in female guinea pigs. AB - In the guinea pig midbrain, a low concentration of progestin receptors is induced by estradiol. This is in contrast to the mediobasal hypothalamus which has a large number of estradiol-induced progestin receptors. Because the midbrain is an important site for the hormonal regulation of sexual behavior, we mapped the distribution of cells containing estrogen receptor- and estradiol-induced progestin receptor-immunoreactivity in that area. Estrogen receptor immunoreactive cells are found in midbrain sites previously reported, including the midbrain central gray, the tegmentum lateral and ventral to the central gray, peripeduncular region, and parabrachial nuclei. While progestin receptor immunoreactive cells were not detected without estradiol priming, estradiol induced progestin receptors were found throughout the rostrocaudal extent of the midbrain central gray and adjacent tegmental area. Progestin receptor immunoreactive cells were far fewer than estrogen receptor containing cells, had less cytoplasmic staining, and appeared restricted to the midbrain central gray, lateral and ventrolateral to the cerebral aqueduct and the adjacent tegmental area. PMID- 8429128 TI - Physiological properties and morphological characteristics of cutaneous and mucosal mechanical nociceptive neurons with A-delta peripheral axons in the trigeminal ganglia of crotaline snakes. AB - Primary A-delta nociceptive neurons in the trigeminal ganglia of immobilized crotaline snakes were examined by intrasomal recording and injection of horseradish peroxidase in vivo. Thirty-four neurons supplying the oral mucosa or facial skin were identified as A-delta nociceptive neurons which responded exclusively to noxious mechanical stimuli and had a peripheral conduction velocity ranging from 2.6 to 15.4 m/s. These neurons were subdivided into a fast conducting type (FC-type) and a slowly conducting type (SC-type). Neurons of both types had a receptive field limited to a single spot which responded to pin prick stimulus with a threshold of more than 5 g. The FC-type neurons had a narrow spike followed by a shorter after-hyperpolarization. In contrast, SC-type neurons exhibited a broad spike with a hump on the falling phase and a longer after hyperpolarization. The diameters of the stem, central and peripheral axons of the FC-type neurons were significantly thicker than those of the SC-type neurons, but there was no statistical difference in the soma size of the two types. Central axons of both types of neurons were thinner than their stem and peripheral axons. Dichotomizing fibers of peripheral axons were observed within the ganglion on 3 neurons. Central axons of the FC-type neurons terminated ipsilaterally in the nucleus principalis, the subnucleus oralis, interpolaris and caudalis and the interstitial nucleus, whereas those of the SC-type neurons generally projected only to the caudal half of the subnucleus interpolaris, subnucleus caudalis and interstitial nucleus ipsilaterally. The present data showed for the first time the physiological and morphological heterogeneity of the primary trigeminal A delta nociceptive neurons and revealed that the trigeminal nucleus principalis and all the subdivisions of the trigeminal descending nucleus are involved in nociception as relay nuclei, but the subnucleus caudalis and the caudal half subnucleus interpolaris are the essential relay sites of the primary nociceptive afferents supplying the oral mucosa and facial skin. The interstitial nucleus also appears to play an important role in orofacial nociception. PMID- 8429129 TI - Possible morphological substrates for GABA-mediated presynaptic inhibition in the lamprey spinal cord. AB - Gamma-aminobutyric acid (GABA) neurons intrinsic to the lamprey spinal cord are known to modulate synaptic transmission from interneurons active during locomotion and from mechanosensory dorsal cells. Many of these physiological effects are presynaptic. To establish the morphological substrates for these axo axonic interactions, an ultrastructural analysis was performed with an antiserum to fixed GABA. The GABA immunoreactivity (ir) was detected by postembedding peroxidase-antiperoxidase and immunogold techniques. GABA-ir terminals were found to make appositions with unlabelled axons located in the dorsal columns and in the ventrolateral aspect of the spinal cord. In the ventrolateral part of the cord, similar appositions between different GABA-ir terminals were also observed. The immunolabelled terminals contained spherical to pleomorphic synaptic vesicles, and also glycogen granules and dense core vesicles. In some cases, the fine structure of the contacts between immunogold-labelled terminals and unlabelled axons suggested a synaptic relationship. Such a relation was found in a relatively small proportion (2-3%) of the appositions studied. These specializations were always observed in close relation to an output synapse of the postsynaptic axon. It is suggested that the axo-axonal contacts described may provide an effective modulation of the synaptic transmission from axons in the lamprey spinal cord. PMID- 8429130 TI - Regenerative outgrowth and distribution of the electroreceptive nerve fibers in the catfish Kryptopterus. AB - The ampullary electroreceptors on the trunk of Kryptopterus bicirrhis are induced and continuously maintained by a trophic substance released by their afferent nerve fibers. When the fibers are transected and the proximal stumps deflected to a new place, the regenerating fibers grow along new routes to the old electroreceptor sites and again induce organs there. The new routes consist of two sections. The first goes from the deflected stump end to other nearby running intact or degenerated nerves and fibers. In the second section, the regenerating fibers grow along these nerves and fibers to their distal ends, i.e., to the old electroreceptor sites. If the path-guiding nerves and fibers are experimentally removed, the displaced fibers grow to the normal receptor areas directly. Growing out from the displaced stump, they spread out over the trunk in a pattern similar to the pattern displayed formerly by the original nerve fibers at this place. It is concluded that the outgrowing fibers do not possess a specificity for a certain receptor site or skin area, but spread out according to epigenetic factors in the periphery. PMID- 8429131 TI - Carbocyanine dye labeling reveals a new motor nucleus in octopus brain. AB - This work aims at a better understanding of the organization of the brain of Octopus vulgaris, emphasizing the touch and visual learning centers. We injected the carbocyanine dye, DiI, into the cerebrobrachial connectives and, separately, into the brachial nerves of living octopuses. In both experiments, retrogradely transported granules of DiI appeared in motor neurons in the superior buccal, posterior buccal and subvertical lobes and in a hitherto unsuspected motor nucleus of several hundred neurons in the posterior dorsal basal and median basal lobes. In addition we labeled afferent fibers by injecting DiI into the caudal (sensory) division of the cerebrobrachial connective on one side; the label spread throughout the superior buccal, posterior buccal and the lateral and median inferior frontal lobes mainly on the injected side. It extended through the cerebral tract into the subvertical lobe, into the superior frontal lobe through the interfrontal tract, through the posterior buccal commissure into the opposite posterior buccal lobe and into the median inferior frontal lobe. The work suggests a new function for the posterior dorsal and median basal lobes, which are shown for the first time to project through the inferior frontal lobe system into the brachial nerves. In addition it represents the first full report of the successful use of the carbocyanine dyes DiI and DiO for labeling nerve tissue in a live invertebrate animal. PMID- 8429132 TI - Astrocyte subtypes in the rat olfactory bulb: morphological heterogeneity and differential laminar distribution. AB - Despite increased recognition of the importance and heterogeneity of astrocyte functions throughout the central nervous system (CNS) relatively little attention has been paid to morphological diversity among astrocytes. Recent studies have indicated that subsets of astrocytes are involved in glial-axonal interactions critical to both development and reinnervation of the rat olfactory bulb. Here, we have characterized the morphologies and distribution of astrocytes within anatomically and functionally distinct layers of the adult main olfactory bulb (MOB). Using a known immunohistochemical marker for astrocytes, glial fibrillary acidic protein (GFAP), and the classic gold sublimate method, we identified six astrocyte subtypes based on their morphology and distribution: (1) unipolar, (2) irregular, (3) wedge-shape, (4) circular, (5) semicircular, and (6) elongate. Unipolar, irregular and wedge-shape astrocytes have not been previously described in the CNS. The unipolar and irregular types are located exclusively in the olfactory nerve layer. Wedge-shape astrocytes are unique to, and are the major subtype in, the glomerular layer. These three morphologically unique astrocyte subtypes may correspond to olfactory nerve layer (ONL) and glomerular layer (GL) astrocytes, which express molecules that regulate axonal growth or synaptogenesis during development and/or regeneration of the olfactory nerve. In the glomerular layer, astrocytes are highly organized with respect to the glomeruli. Individual astrocytes are loyal to a single glomerulus. In the external plexiform layer, astrocytes are spaced relatively uniformly. In the granule cell layer, astrocytes appear to compartmentalize granule cell aggregates, recently shown to be coupled by tight junctions. The distribution and patterns of astrocyte processes and the density of GFAP immunoreactivity are distinctive for each of the layers of the olfactory bulb. The spacing of astrocytes and the organization of their processes may be important to compartmentalization of neuronal functions. High levels of GFAP immunoreactivity correlated with layers of high neuronal plasticity. The morphological diversity and differential distribution of astrocytes in the olfactory bulb reported here support growing evidence for functional diversity of astrocytes and important interactions among specific astrocyte and neuron subtypes. It is reasonable to hypothesize, therefore, that as for neurons, morphologically distinctive astrocyte subtypes may correspond to functionally specific classes. PMID- 8429133 TI - The diencephalon of the Pacific herring, Clupea harengus: cytoarchitectonic analysis. AB - The cytoarchitecture of nuclei in the preoptic area, ventral thalamus, dorsal thalamus, epithalamus, hypothalamus, posterior tuberculum, synencephalon, and pretectum and the accessory optic nuclei was analyzed in the clupeomorph teleost, Clupea harengus. Plesiomorphic (evolutionarily primitive) and apomorphic (evolutionarily derived) features of nuclei were identified by cladistic analysis. Plesiomorphic features include the cytoarchitectonic organization of most of the preoptic nuclei, the somewhat scattered cells of nucleus ventrolateralis, the compact, oval shape of nucleus intermedius, the presence of dorsoventrally oriented laminae in the central posterior nucleus, and most features of the hypothalamic nuclei. Also plesiomorphic are the presence of a thick, prominent paraventricular organ, a nucleus of the paraventricular organ, a nucleus tuberis posterior, and a preglomerular complex in which the boundaries between multiple nuclei are relatively difficult to distinguish. Additionally, the cytoarchitecture of the three synencephalic nuclei present in Clupea, the presence of small cells in nucleus pretectalis superficialis pars parvicellularis and of larger, scattered cells in nucleus pretectalis superficialis pars magnocellularis, the presence of large cells in the dorsal accessory optic nucleus that form a rostrocaudally oriented column, and the feature of a small, cell-sparse ventral accessory optic nucleus are plesiomorphic. Apomorphic features include the presence of a single, large, circular lamina that surrounds a central neuropil in all but the most caudal part of nucleus anterior, a lack of bilateral asymmetry in the habenular nuclei, the relatively small size of the periventricular nucleus of the posterior tuberculum, the presence of two, distinguishable caudomedial nuclei in the posterior tuberculum, elongation and folding of the neuropil of nucleus pretectalis superficialis pars parvicellularis, and the relatively large size of nucleus pretectalis superficialis pars magnocellularis and the posterior pretectal nucleus. PMID- 8429134 TI - Principal neurons as local circuit neurons in the rat superior cervical ganglion: the synaptology of the neuronal processes revealed by intracellular injection of biocytin. AB - To analyze the local circuitry of the sympathetic ganglion, the synaptic relations of the neuronal processes of the principal neurons in the rat superior cervical ganglion were investigated by correlated light and electron microscopy combined with intracellular injection of biocytin. Intracellular iontophoresis of biocytin followed by avidin-biotinylated horseradish peroxidase cytochemistry allowed complete visualization of the neuronal processes of the principal neurons. The stained principal neurons have a single process (axon), which leaves the ganglion, and several intraganglionic processes (dendrites), some of which show specific terminal arborizations. Some terminals of the dendritic collaterals formed pericellular plexuses or intercellular glomerular plexuses. Electron microscopically, the dendrites and their collaterals contain numerous small vesicles. Synaptic membrane specializations were observed between the stained dendritic collaterals and unlabeled neurites. These may be both preganglionic axon terminals and processes of principal neurons. The likely direction of neurotransmission often could not be determined because of the bidirectional synaptic structures. Our findings show that the dendritic collaterals of principal neurons appear to make both post- and presynaptic contacts with both the principal neurons and the preganglionic axons. It is suggested that the principal neurons might participate in local circuits involving not only preganglionic axons but also neighboring principal neurons. PMID- 8429135 TI - Skin trapping. PMID- 8429136 TI - Effectiveness of the health campaign for the early diagnosis of cutaneous melanoma in Trentino, Italy. AB - BACKGROUND: To reduce the mortality rate from cutaneous melanoma in Trentino, a health campaign for early diagnosis aimed at the general public was organized. OBJECTIVE: The mortality rate from cutaneous melanoma in Trentino before and after our health campaign was analyzed and compared with the mortality rates in three neighboring regions where no campaigns were carried out. METHODS: The mortality for cutaneous melanoma was analyzed using data from the death certificates provided by ISTAT for the pre- and postcampaign periods. The two summary methods: cumulative rate and the standard mortality ratio were used. RESULTS: The cumulative mortality rates in the neighboring regions show a tendency to increase over the three periods whereas the increase in Trentino is less evident. The standard mortality ratios confirmed this result. CONCLUSION: We feel that this type of campaign is to be recommended in the reduction of mortality from cutaneous melanoma. PMID- 8429137 TI - Intralesional cryosurgery. A new technique using cryoneedles. AB - A new method for applying cryosurgery in depth has been performed in a trial in order to achieve more effectiveness and avoid many of the disadvantages of conventional techniques. A needle is introduced into the skin from one point and runs through the deeper tissues of the lesion, appearing at the surface on the opposite border. A cryogen is then passed through one end of the needle and vents to the atmosphere from the opposite end. An ice cylinder is formed around the embedded part of the needle within the deeper tissues. The distance of extension of freezing can be estimated by insertion of thermocouple needles as well as by the degree of extension of the whitish ice balls formed around the points of contact between the skin surface and the visible portions of the needle. New varieties of needles have been developed to facilitate improved efficacy in achieving better depth of freeze. The needles are angled, curved, and hook shaped. Some are partially insulated to better localize the direction of freezing. The process was very practical, and effectively eradicated 85% of epidermal lesions after one freeze; 15% needed additional sessions as did deeper dermal lesions. PMID- 8429138 TI - Role of tissue undermining in the trapdoor effect of transposition flaps. AB - BACKGROUND: The trapdoor or pincushioning effect is a frequent complication of transposition flaps. Several explanations have been proposed for its occurrence, including lymphatic or venous obstruction, scar hypertrophy, excessive subcutaneous fat or flap tissue, and scar contracture. OBJECTIVE: To study the effects of tissue undermining and scar contracture using a guinea pig animal model. METHODS: Circular wounds on the dorsal surface of guinea pigs were repaired with transposition flaps. Half of the recipient beds were undermined widely and half were not undermined. Animals were observed for evidence of the trapdoor phenomenon. RESULTS: Only animals in the group without undermining demonstrated evidence of the trapdoor effect. CONCLUSION: Tissue undermining may prevent the development of the trapdoor effect in transposition flaps. PMID- 8429139 TI - Basal cell carcinoma of the male nipple. Case report and review of the literature. AB - BACKGROUND: Basal cell carcinoma (BCC) of the nipple is a very unusual occurrence, often mistaken for Paget's disease. OBJECTIVE: By reporting a further case of BCC of the nipple in a 57-year-old man and by reviewing the sixteen previously reported cases, we mean to characterize further a common lesion arising at an unusual site. We also wish to stress the lack of agreement regarding the treatment of BCC of the nipple. METHODS: Review of the literature was based on a search in Medline. CONCLUSIONS: We appeal to compile all cases of BCC arising at unusual locations in order to gain a better understanding of these lesions. PMID- 8429140 TI - Full-thickness mucous membrane grafts from the vestibular (oral labial) mucosa of the lip. AB - BACKGROUND: Full thickness mucous membrane coverage is desired for a variety of reconstructive tasks involving the oral cavity and vermilion area of the lips. OBJECTIVE: This article will inform the readership that the vestibular (oral labial) mucosal area of the lower lip close to the sulcus may provide adequate full-thickness mucous membrane graft tissue for such purposes. METHOD: A series of photographs from a number of representative cases with accompanying commentary will enable the surgeon to easily incorporate this technique into his/her practice. CONCLUSION: The lips vestibular mucosa will be found to be an invaluable ally in accomplishing non-deforming coverage of superficial mucosal defects without unnecessary surgical invasion of visible lip/chin structures. PMID- 8429141 TI - Establishing the physician-patient relationship. AB - BACKGROUND: This is the first in a series of articles written to address the many medicolegal issues that impact the practice of dermatology. The topic of establishing the physician-patient relationship was chosen as the starting point since it is the cornerstone of every medical malpractice action. METHODS: A format of an initial discussion of legal concepts followed by hypotheticals and a review of actual cases was chosen to enhance interactive learning. The subject matter is based on a review of medicolegal literature and actual recorded case law. RESULTS: A physician-patient relationship may be established whether or not there is actual physical contact between the physician and patient, there is gratuitous advice, or there is an academic purpose. CONCLUSION: The key in determining whether a physician-patient relationship has been established is whether the patient reasonably relied upon advice given or had an expectation of treatment. PMID- 8429142 TI - Basal cell carcinoma arising in venous ulcers and mimicking granulation tissue. AB - BACKGROUND: Basal cell carcinoma (BCC) arising in venous ulcers is rare, and its appearance in this setting is not fully recognized. OBJECTIVE: The purpose of this report is to alert the clinician to the subtle clinical appearance of this tumor and to prompt a more rapid recognition. METHODS: We report five cases of BCC arising in venous ulcers of the lower extremity that we have observed over a period of 6 years. In all cases the tumor was diagnosed histologically. RESULTS: In all five cases the tumor had the appearance of seemingly healthy granulation tissue, which was often exuberant and translucent. The border of this apparent "granulation tissue" was rolled over the margins of the ulcer. CONCLUSION: Basal cell carcinoma arising in venous ulcers often has the appearance of translucent granulation tissue that extends beyond the ulcer's margins. These clinical characteristics should alert the clinician to the need for obtaining a biopsy of the ulcer bed. PMID- 8429143 TI - Angiosarcoma of the scalp treated with Mohs micrographic surgery. AB - The idiopathic type of angiosarcoma most commonly occurs on the face and scalp of elderly persons. This rare tumor can be highly aggressive; however, if the diagnosis is made at an early stage, while the tumor is still localized and has a size that is less than 10 cm, surgical excision is the treatment of choice. We report a case whereby Mohs micrographic surgery was used to treat angiosarcoma of scalp. PMID- 8429144 TI - Recurrent malignant melanoma following a 35-year disease-free interval. AB - A case is presented of recurrent melanoma following a 35-year disease-free interval after treatment for stage I disease. Although the case seems to indicate a recurrence of melanoma, the possibility that the "recurrent lesions" may have been a new primary lesion should be considered. Regardless of disease-free interval following initial treatment, a complete cure of the disease can never be assumed. PMID- 8429145 TI - Injectable collagen and autoimmune disease. AB - BACKGROUND: The media and litigation in the United States have alleged a link between the use of injectable bovine collagen and the development of autoimmune disease, specifically dermatomyositis/polymyositis. OBJECTIVE: To present the scientific and epidemiologic data with regard to this issue. METHODS: Literature was reviewed as well as reports to Collagen Corporation and the litigation. RESULTS: After review of the literature and review of data currently available to Collagen Corporation, no link can be established at this time between collagen injectable material and autoimmune disease, specifically dermatomyositis/polymyositis. CONCLUSION: There are no epidemiologic or scientific data to support the alleged link in the media and litigation between injectable bovine collagen material and dermatomyositis/polymyositis. PMID- 8429146 TI - Punch biopsy: one more possible stage in Mohs micrographic surgery. PMID- 8429147 TI - Forehead flap donor site closure. PMID- 8429148 TI - The "wrinkle test": clinical use for detecting early epidermal resurfacing. PMID- 8429149 TI - The Acu-guard sterile disposable electrosurgery cover. PMID- 8429150 TI - Closure of wounds under tension with the pulley suture. PMID- 8429151 TI - Misrepresentation of minigrafts. Fees and protocol. PMID- 8429152 TI - Recurrence rates of treated basal cell carcinomas. PMID- 8429153 TI - An ethical responsibility for pain management. PMID- 8429154 TI - Asthma patients' knowledge in relation to compliance with drug therapy. AB - Patient knowledge of asthma and treatment and compliance levels were assessed among 100 moderate to severe asthmatics recruited from general practice. With the use of postal questionnaires non-compliance was found to be high: 39% of patients omitted to take their asthma treatment as prescribed. The level of patient knowledge had no significant effect on compliance to drug therapy. The highest compliers were respondents who reported never receiving an explanation about the condition. The majority of patients believe they would know how to manage an attack, but when they 'scored' on their ability only 34.4% were deemed to be safe. However, the level of patient knowledge appears to influence a patient's ability to manage an asthma attack. Less than half of the patients who had had asthma explained to them reported to have understood the initial explanation, and explanations made by nurses were particularly poorly understood. The study identifies reasons why patients do not comply with drug treatments. Many feel the drugs are not necessary and many forget to take them. Almost half of the cohort admit a reluctance to use their inhalers in public and a third state a preference for tablets rather than inhalers. Health professionals must look at other means of improving patient compliance rather than education in isolation. Since the general practitioner contract was introduced in April 1990, nurses working in general practice have become increasingly involved with health education as part of health promotion and chronic disease management clinics. This study highlights the need for further education in this area. PMID- 8429155 TI - Assessment of competence in clinical practice: a phenomenological approach. AB - Using a phenomenological approach, this study explores the different dimensions of student nurse assessment in clinical practice from the perspective of 10 experienced practitioners. Phenomenology, one of the various types of qualitative research methods, is used to discover and understand the meaning of human life experiences through an analysis of the subjects' descriptions of situations. It is therefore of particular value when addressing the questions posed in the present study: (a) what attributes characterize competence and non-competence in learner nurses, (b) how they recognize these attributes in practice, and (c) how they measure competence at different levels of preparation. Rather than provide answers, the phenomenologist explores the meaning of the language used in determining competence in practice. Through this contribution to the understanding of the notion of competence and its measurement by these experienced practitioners, it may be possible in some way to unravel the confusion that surrounds the meaning of competence represented in the literature. It is hoped that, through the discovery of the lived world of these clinicians, others may be better facilitated for and within their role as assessors in the future. PMID- 8429156 TI - The Patient-Centredness Multi-Choice Questionnaire: developing an instrument for the measurement of patient-centredness in student nurses. AB - An earlier paper by the writer reported on the construction and administration of an instrument for the measurement of therapeutic attitudes in student nurses. This paper continues that work by producing a short, appropriate and easily administered test, known as the Patient-Centredness Multi-Choice Questionnaire. The instrument was tested for validity and reliability, and was piloted on a large group of new Project 2000 students from several colleges and institutes of nursing, to produce a database for use in further studies. PMID- 8429157 TI - The development of a measure of job satisfaction for use in monitoring the morale of community nurses in four trusts. AB - The development of the Measure of Job Satisfaction (MJS) for use in a longitudinal study of the morale of community nurses in four trusts is described. The review of previous studies focuses on the use of principal component analysis or factor analysis in the development of measures. The MJS was developed from a bank of items culled from the literature and from discussions with key informants. It was mailed to a one in three sample of 723 members of the community nursing forums of the Royal College of Nursing. A 72% response rate was obtained from those eligible for inclusion. Principal component analysis with varimax rotation led to the identification of five dimensions of job satisfaction; Personal Satisfaction, Satisfaction with Workload, Satisfaction with Professional Support, Satisfaction with Pay and Prospects and Satisfaction with Training. These factors form the basis of five subscales of satisfaction which summate to give an Overall Job Satisfaction score. Internal consistency, test-retest reliability, concurrent and discriminatory validity were assessed and were found to be satisfactory. The factor structure was replicated using data obtained from the first three of the community trusts involved in the main study. The limitations of the study and issues which require further exploration are identified and discussed. PMID- 8429158 TI - Nursing research conventions: objectivity or obfuscation? AB - This paper is a critique of naive realism, the philosophy which animates much nursing research, and which leads researchers to assume that the attainment of objective knowledge is possible. The nature of naive realism, and its relationship to objectivity, is discussed. Central to this outlook is the belief that the values and interests of the researcher can and should be divorced from the prosecution of research. This is reflected in the literary convention of referring to the researcher in the third person. Contrary to this position, I argue that the interpretations, values and interests of the researcher are central to the research process. Moreover, nursing research may be affected by the interests of managers, educationalists, and those who wish to see nursing attain professional status. Nursing researchers should accept that they are part of the social situations which they study. They should therefore become reflexive in their outlook. This entails recognizing and attempting to understand the effects of the researcher, rather than trying to eliminate or ignore them. PMID- 8429159 TI - Interactions of nurses with elderly patients: relationship to nurses' attitudes and to formal activity periods. AB - Two different strategies are possible for improving nurses' interactions with elderly patients. One is to arrange special periods, such as reality orientation, in which a high level of appropriate interaction is encouraged. The other is to improve nurses' attitudes to patients and treatment, or select nurses with better attitudes, on the grounds that poor attitudes might underly poor behaviour. This study was designed to find out whether nurses' interactions with elderly patients varied in relation to their attitudes, or differed between periods of routine care or of special activities based on reality orientation organized by the nurses. Systematic observations were made of the behaviour of 27 nurses working on two psychogeriatric wards, before all nurses completed questionnaires measuring attitudes to nursing care and to elderly people. A higher proportion of interactions were 'positive' (i.e. involved nurses' informing, questioning or making general conversation) during activity periods than at other times. No attitudinal measure predicted the proportion of interactions which were positive, although this was greater in qualified nurses. The results are consistent with the view that formal reality orientation periods can improve the quality of nurses' interactions with elderly patients, but are inconsistent with the view that targeting nurses' attitudes would be effective. PMID- 8429160 TI - A multi-generational staff development model for registered nurses. AB - The paper focuses on a significant nursing initiative for staff development undertaken in South Australia. The initiative arose out of Health Commission funded consultancy which sought to develop a staff development model for registered nurses. The outcome of this consultancy was the development of a multi generational staff development model, known as the DPS Staff Development Model. This model provides a framework through which staff development activities can be analysed, consolidated and extended. The paper outlines the DPS Staff Development Model, the context in which it was developed and its implications for nursing staff development programmes. The development of the model was premised on a number of assumptions. These included the need to ensure flexibility, longevity, offer a perspective of staff development that is realistic and practical, and facilitate in registered nurses a sense of ownership and personal responsibility with respect to their development. The model comprises six broad areas of focus: the Career Development Model for registered nurses in South Australia, the structure of the model, locations in which staff development occurs, elements which assist in maintaining skills and knowledge, elements which enhance growth and development and the processes and relationships necessary for the successful implementation of staff development programmes. PMID- 8429161 TI - The emerging role of the British nurse teacher in Project 2000 programmes: a Delphi survey. AB - The findings of a three-round Delphi survey, conducted to explore the role of the British nurse teacher as it emerges within Project 2000 programmes, are discussed. The panel of experts comprising 201 grade 2 nurse teachers was drawn from the 25 of the 28 colleges in England who had implemented Project 2000 between September 1989 and April 1991. The findings highlight the complex and multifaceted role of the nurse teacher and the changes which are occurring in the role. Currently, the main changes are related to teaching and learning activities. These emphasize the depth, level and specialist nature of the teaching, and teaching on a variety of courses at diploma and degree level. Assessment activities are given the highest overall rating as important, and recruitment activities the lowest, in both the current and future roles. Educational management and administrative activities are found to be numerous and currently do not appear to be declining. The few changes that have occurred to the administrative areas have been replaced with similar activities. Another important area for the future was perceived to be research activity. In view of the developing partnership with higher education in the United Kingdom, this is not a surprising finding. PMID- 8429162 TI - Dame Muriel Powell DBE, 1914-1978. PMID- 8429163 TI - Quality of care in nursing homes: from the resident's perspective. AB - This paper derives from a study conducted by the Deakin Institute of Nursing Research between 1988 and 1990, whose major objective was to determine the impact of staffing mix on nursing resident's quality of care and life. Resident satisfaction with life in the nursing home is a key element in determining the quality of care and quality of life provided. Both the literature review and the study objectives supported the view that resident outcome can be collected through assessing the quality of care and the quality of life, through assessment by informed observers using instruments derived from explicitly stated standards, and through eliciting the perceptions of residents themselves. A schedule designed to measure satisfaction with care was developed and resident interviews were undertaken using this measure and the Life Satisfaction Index (A). The majority of responses to the resident satisfaction schedule were positive. The high percentage of positive responses did not correlate with the observations of the research assistants and there was some concern that while residents were able to assess care they were reluctant to criticize the staff or their behaviour. PMID- 8429164 TI - Measuring feeding difficulty in patients with dementia: perspectives and problems. AB - The feeding problems of demented elderly people are well documented and the need for research into the assessment of feeding difficulty and intervention by nurses has been raised. The present paper reviews the literature in this area of care and outlines the problems which exist in attempting to investigate the feeding difficulty of demented patients. Demented elderly people display a range of behaviours related to feeding including excessive eating in the early stages of dementia and then difficulty with feeding, refusal to eat and, finally, inability to self-feed at all. The problems with research in this area revolve around the issue of measurement of feeding difficulty. There are problems in deciding what to measure and in how measurements should be made which are clinically meaningful. A possible strategy for investigating the feeding difficulty of demented elderly patients is suggested which includes the design of a tool for measurement and the application of single-case studies. Any tool which is used for measurement should enable researchers, in the first instance, and then clinicians to categorize the feeding difficulty of individual patients. Moreover, such a tool should also be sufficiently sensitive to respond to change in feeding ability. The single-case methodology is considered to be the most ethically and statistically appropriate for research with this particular group of patients. PMID- 8429165 TI - Quality of life: a concept analysis. AB - Quality of life (QOL) is a phrase which was first used shortly after the Second World War and has, since then, been overused and infrequently defined. A concept analysis of QOL is presented to clarify the concept for further use. The process for concept analysis developed by Walker & Avant is employed. Quality of life is found to be very complex, and it is hoped that this analysis will stimulate thought and further nursing research into what QOL means in the health care context. PMID- 8429166 TI - Head-injured survivors: caregivers and support groups. AB - The purposes of this study were to assess the needs of caregivers of head-injured survivors and the availability, use and helpfulness of support groups in meeting these needs. Demographic data on characteristics of the caregivers and of the head-injured survivor were also collected. The majority of the caregivers were mothers of head-injured males and provided care for an average of 6 years. Sixteen of the respondents did not attend a support group, mainly due to the unavailability of support groups in their area of residence. Support groups were most helpful in meeting educational and psychological needs of caregivers. The findings suggest the need for additional support groups, especially in smaller communities, for additional educational content in support groups and for respite beds. PMID- 8429167 TI - Common adaptive tasks facing parents of children with chronic conditions. AB - Parents of chronically ill or disabled children face a number of common tasks in adapting to their child's condition. These tasks are delineated from the literature on theory, research and clinical issues and are discussed in relation to their documented importance to family adaptation, the difficulties parents have in managing the tasks and effective coping strategies parents can use to accomplish the tasks. The implications of utilizing adaptive tasks as an organizing framework for assessing and intervening with parents of children with chronic conditions are discussed. PMID- 8429168 TI - Advice concerning breastfeeding from mothers of infants admitted to a neonatal intensive care unit: the Roy adaptation model as a conceptual structure. AB - Data were collected by telephone interviews with 178 mothers of full-term patients in a NICU (neonatal intensive care unit) concerning advice on facilitation of the initiation of breastfeeding. The main advice to the first author as a nurse in the NICU concerned the environment, advice on breastfeeding, distance between units, work organization and nurse behaviour. The advice to other mothers of patients centred on persistence, physical contact with the infant, and not to let nurses take over maternal role functions. The data were structured into themes and categories, classified by one author and two research assistants according to Roy's adaptation theory, and analysed for degree of interrater agreement. The overall agreement of classification was high, reaching 92.5%. It was easily applied by nurses after a brief introduction and proved useful for structuring interview data. It also contributed to clarification of nurse behaviour and division of roles between nurses and mothers. As the four adaptation modes showed considerable overlap, this kind of classification seems inadvisable for application to the assessment of patient/parent situations in the nursing process. For use in a clinical setting, the theory needs the addition of the interactive aspect of nurse and patient/family role functions, and may then be used as a framework for the development of assessment tools. PMID- 8429169 TI - How do nurses assess the information received by breast cancer patients? AB - The aim of the study was to clarify how nurses assess the information targeted at breast cancer patients before, during and after hospitalization. The sample group consisted of 176 nurses from surgical and radiological wards and clinics in southwest Finland. The study used responses to a questionnaire investigating breast cancer patients' recovery. Of the sample, 71% responded to the questionnaire. Log-linear modelling was used for the final statistical analysis of the results. The findings obtained by log-linear modelling showed a cumulative effect. Various associations were seen in the responses of the nurses on how they assessed the information received by patients. Patients who were considered well informed before hospitalization were similarly considered during their hospital stay. Nurses consider that they give their patients insufficient information. Further, nurses are not clear about their own role in the distribution of information. PMID- 8429170 TI - Evaluation of services for people with HIV/AIDS in an inner-city health authority: perspectives of key service providers. AB - This paper reports the findings of the first phase of a study designed to assess the needs of people with HIV and AIDS and the extent to which these needs are being met by services in Camberwell Health Authority in inner London. The study involves collecting information and opinions from key service providers, clients with HIV/AIDS and their informal carers. It is the service provider data which will be presented here. Semi-structured interviews were conducted with 47 key service providers involved in the care and support of people with HIV/AIDS in the Camberwell Health District. Respondents were drawn from the local health services, local social services departments and a number of locally active voluntary organizations. The aim was to gather respondents' views on their individual roles and the roles of the organizations they represented, the health and social needs of people with HIV/AIDS and the extent to which these were being met by current service provision and the co-ordination of services for people with HIV/AIDS. The data showed that a broad range of services were utilized by people with HIV/AIDS, reflecting a wide variety of health and social care needs. The majority of respondents felt that some of the needs of people with HIV/AIDS were similar to those of people living with other chronic illnesses (such as cancer). However, they also identified a number of problems which were either unique or more severe for people with the virus. Services for people with HIV/AIDS were generally felt to be poorly co-ordinated and a number of areas of overlap and gaps in service provision were identified. PMID- 8429171 TI - Coping strategies and health status of elderly arthritic women. AB - With increasing age many elderly people experience personal and social losses. Physical health declines, there is a loss of vigour and an increased susceptibility to disease. Osteoarthritis, the most common form of arthritis in the elderly, is the greatest cause of disability and limitation of activity in older populations. The purpose of this descriptive study was to investigate how elderly women cope with the physical impairment and pain associated with osteoarthritis and to examine the relationship between coping strategies and health status. The sample consisted of 130 women over 75 years, living alone in one of five life care communities. Three instruments were used to collect data: The Ways of Coping Scale, The Arthritis Impact Measurement Scale and The Musculoskeletal Impairment Index. Pearson product-moment correlations and stepwise multiple regression were used to analyse data. The mean age of the sample was 83.2 years and over 40% had attained bachelor's degree or higher. The most utilized coping strategy was self-control. The study variables accounted for 49% of the variance in physical health and 20% in psychosocial health. PMID- 8429172 TI - AIDS-related experience, knowledge, attitudes and beliefs amongst nurses in an area with a high rate of HIV infection. AB - The results are presented of a survey of the AIDS-related knowledge, attitudes and beliefs of a random sample of 600 qualified nurses in the Lothian Region of Scotland. This locality is characterized by a high rate of detected HIV infection which is mainly associated with intravenous drug use. This study indicated that 64% of respondents had contact with HIV-infected patients and that 48% had nursed people with AIDS. Levels of AIDS-related knowledge were not high and a substantial proportion of nurses reported lacking basic AIDS information and professional support to enable them to work effectively with people who have AIDS. PMID- 8429173 TI - Personal unity in dying: alternative conceptions of the meaning of health. AB - Death is a universal phenomenon. In the health care system, however, it is regarded as unfortunate and catastrophic. Death is perceived as the extreme end of a continuum whose opposite end is health. In nursing practice, through the application of nursing diagnosis and nursing process, dying is treated as a procedural problem. Rarely is dying treated as part of the experience of living, characterized by the meanings assigned by the individual to the event. The purpose of the study was to describe one person's dying experience as a personal expression of the meaning of health. PMID- 8429174 TI - Euthanasia: a comparison of the lived experience of Chinese and Australian palliative care nurses. AB - This paper reports on a study which investigated whether nurses from Eastern and Western cultures hold differing definitions of euthanasia and different perspectives of the effect of their attitudes to euthanasia on professional relationships with patients and colleagues. The focus of the study was the lived experience of Chinese and Australian nurses working in palliative care. The results indicate that there were differences between these two groups in their definitions of euthanasia. Lived experience obtained from both groups revealed conflict with patients and colleagues, and a lack of opportunity to discuss the ethical issue of euthanasia in the workplace. PMID- 8429176 TI - The cracked tooth syndrome: a pragmatic treatment approach. PMID- 8429175 TI - Amalgam restorations. PMID- 8429177 TI - The clinical diagnostic process: a prerequisite to excellence in practice. AB - The ability to render good clinical diagnosis represents the mark of the "healing arts" professional. A sequence of diagnostic steps is proposed. Adherence to this scheme should be helpful in reducing diagnostic failures, thus improving patient care. PMID- 8429178 TI - Sealants and handpieces. PMID- 8429179 TI - NIDR research offers new therapies for arthritis, periodontal disease. PMID- 8429180 TI - Surgeon general warns against use of smokeless tobacco. PMID- 8429181 TI - Shedding new light on lasers. Some timely words of caution for readers. PMID- 8429182 TI - Lasers in dentistry: an overview. AB - Developments in laser dentistry have led to an increasing acceptance of this technology by both practitioners and the general public. This article provides an overview of laser systems and their dental applications. PMID- 8429183 TI - Using lasers in clinical dental practice. AB - Lasers are an impressive potential treatment modality for a variety of clinical conditions. This article profiles several clinical applications in dentistry. PMID- 8429184 TI - Lasers in dentistry: comparing wavelengths. AB - Confusion exists about the wavelengths marketed for dental use. General properties of lasers, laser/tissue interactions and clinical applications of three wavelengths are examined. PMID- 8429185 TI - Laser/tissue interaction: what happens to laser light when it strikes tissue? AB - Several factors influence laser/tissue interactions, and various combinations of these factors could damage tissues. This article examines where laser energy goes when it strikes tissue and how it effects tissues during laser treatment. PMID- 8429186 TI - The effect of lasers on dental hard tissues. AB - There is a quest to find a method to remove diseased and healthy dental hard tissues without the negative stimuli associated with dental handpieces. Today, lasers are being considered as a potential replacement. This report evaluates effects of three lasers on dentin and pulpal tissues. The Er:YAG laser appears to have a lesser thermal effect. PMID- 8429187 TI - Recent developments in restorative dental ceramics. AB - Since the introduction of porcelain jacket crowns in the early 1900s, dental ceramics have been praised for their esthetic appearance. But there's more to consider: other benefits, longevity, limitations. This review addresses major developments in modern dental ceramics and metal-ceramics. PMID- 8429188 TI - Is there life after dentistry? PMID- 8429189 TI - Assessing patient pain during dental laser treatment. AB - Patients reported little or no pain during or after treatment with a dental laser. With many people reporting fear of pain as their chief reason for not seeking dental care, lasers may offer a more acceptable treatment technique. PMID- 8429190 TI - Patient self-determination act. Implications for long-term care. AB - The Patient Self-Determination Act requires providers to give all new nursing facility residents information about right-to-die and other treatment options under state law. Most patients expect family members to make key decisions, or believe it is the physician's responsibility. Simply handing out material is unlikely to have any impact on the number of patients completing advance directives. Even with advance care directives in place, such end-of-life desires are often not carried out. PMID- 8429191 TI - Finnish nursing homes: client well-being & staff development. AB - This study concerned the changes in the physical and mental problems experienced by patients before and after a staff education program on an experimental ward in a Finnish nursing home. It was difficult to conclude, on the basis of significant differences, whether the development program decreased the problems of patients. The findings, however, demonstrated that the patients on the experimental ward were significantly less anxious and dissatisfied after the program. It can be difficult to demonstrate significant differences in the outcome of "old" and "new" nursing practices, but changes may occur in many areas that the elderly can experience qualitatively but which cannot be expressed quantitatively. PMID- 8429192 TI - Hospitalized elders. Risk of confusion with hip fracture. AB - Although frequently underdiagnosed, more than half of all patients admitted for the treatment of hip fracture are confused at the time of hospital admission or develop confusion sometime during the course of hospitalization. This study reveals the profound vulnerability of severely confused patients with hip fracture. Severely confused patients experience higher overall rates of medical complications and are more likely to be discharged to a nursing home. In addition to being at higher risk for medical complications, severely confused patients also appear to be at greater risk for being physically restrained. PMID- 8429193 TI - Special care units for dementia. PMID- 8429194 TI - Interpretation of abnormal laboratory values in older adults: Part II. PMID- 8429195 TI - Music debate continues. PMID- 8429196 TI - Urinary retention in hospitalized elderly women. AB - Unrecognized urinary retention may be a factor in the development of recurrent urinary tract infections, urinary incontinence, bladder dysfunction, and upper urinary tract disease. Because urinary retention is often asymptomatic in elderly women, with amounts of up to 1,500 mL retained in the bladder often causing little or no discomfort, nurses need to have a high index of suspicion about its occurrence. A post-void residual is the best way of determining the presence of urinary retention. The goal of treatment for urinary retention is bladder decompression via catheterization followed by voiding trials and determination of residual volumes. PMID- 8429197 TI - Co-localization of erythropoietin mRNA and ecto-5'-nucleotidase immunoreactivity in peritubular cells of rat renal cortex indicates that fibroblasts produce erythropoietin. AB - In adults, the kidneys are the major site of production of the glycoprotein hormone erythropoietin (EPO), but the type of renal cell producing EPO has not yet been identified. In the present study we used non-radioactive in situ hybridization with a digoxigenin-labeled cRNA probe to localize cells that produce erythropoietin (EPO) in kidneys of anemic rats. Cryostat sections from both native and perfusion-fixed tissue were used. Cells containing EPO mRNA were found exclusively in the peritubular space of the renal cortex. Using high resolution interference contrast optics, we found that cells expressing EPO mRNA were not associated with the lumina of peritubular capillaries but rather were located in the angles between adjacent tubules or between tubules and vessels. These spaces are predominantly occupied by resident interstitial fibroblasts and by their cytoplasmic processes. To further identify the type of cell containing EPO mRNA, a double-labeling protocol was established that permitted on the same tissue section both in situ hybridization for EPO mRNA and parallel immunolabeling of ecto-5'-nucleotidase (5'-Nu), a surface marker of peritubular interstitial fibroblasts. The combined labeling technique revealed that a clear majority of cells expressing EPO mRNA also displayed staining for anti-5'-Nu. Staining for EPO mRNA was localized in central perinuclear parts of the interstitial cells, whereas 5'-Nu label was present on the cell surface, including the cytoplasmic processes. These data indicate that peritubular fibroblasts are cellular sites for production of erythropoietin. PMID- 8429198 TI - Immunoelectron microscopy of carbonic anhydrase isozyme VI in human submandibular gland: comparison with isozymes I and II. AB - Carbonic anhydrase VI (CA VI) was purified from human saliva by inhibitor affinity chromatography, and its distribution was studied in human submandibular gland by the indirect immunoperoxidase technique with a rabbit polyclonal antibody raised against the isozyme. Polyclonal antibodies to human CA I and CA II purified from erythrocytes were also raised and used for immunostaining. SDS polyacrylamide gel electrophoresis of the purified isozymes revealed a single protein band (CA VI, 42 KD; CA I and CA II, 30 KD). Antibody raised against CA VI did not crossreact with CA I or CA II either by Western or by dot-blotting. However, antibodies against CA I and CA II showed slight crossreaction with each other's antigen by dot-blotting. In a Western blot of purified submandibular gland CA, antibody to CA VI stained the 42 and 30 KD bands, and antibodies to CA I and CA II stained the 30 KD band. The 42 KD but not the 30 KD molecule was cleaved by endo-beta-N-acetylglucosaminidase F, indicating that the former contains N-linked oligosaccharides. Immunostaining for CA VI was seen in the secretory granules and cytosol of serous acinar cells and in the duct luminal contents. Staining specific for CA II was observed in the cytosol of serous acinar and duct epithelial cells. Antibody to CA I reacted only with the walls of small blood vessels. These results suggest that (a) serous acinar cells secrete 42 KD CA VI which functions in the oral cavity and that (b) serous acinar and duct epithelial cells possess cytosolic CA (30 KD CA VI and CA II) which functions in situ. PMID- 8429199 TI - Renin processing in cultured juxtaglomerular cells of the hydronephrotic mouse kidney. AB - We examined renin processing in cultured juxtaglomerular (JG) cells of the hydronephrotic mouse kidney with immunocytochemical and biochemical techniques. Compared with JG cells in normal kidneys, there was less intense labeling for renin protein in mature granules of cultured JG cells. However, pro-renin labeling of transport vesicles and juvenile granules was maintained, suggesting incomplete passage of pro-renin through intermediate and mature granules. Immunogold evidence of exocytosis of mature granules containing renin protein was present at all stages. Labeling of transport vesicles for pro-renin, together with the absence of exocytosis of pro-renin from juvenile granules, indicated that pro-renin was exclusively released by a constitutive process. Active renin release into supernatants decreased with time, whereas the ratio of total renin to active renin increased, indicating that pro-renin synthesis and release were maintained but that the processing of pro-renin to active renin was interrupted. Angiotensin II inhibited and verapamil stimulated active renin release in culture; neither substance affected pro-renin release. Application of secretagogues that act via intracellular calcium or cAMP resulted in depletion of mature granules and their deformation by myelin figures and vacuoles, findings consistent with an exocytosis from mature granules. The absence of effect of any secretagogues on pro-renin release suggests that these stimulatory mechanisms are exclusively post-Golgi. In cultured JG cells in renal explants, renin vesicular transport and granular exocytosis are maintained but a defect in pro-renin passage from juvenile to intermediate granules is apparent. PMID- 8429200 TI - Scanning electron microscopy of type I collagen at the dentin-enamel junction of human teeth. AB - The dentin-enamel junction constitutes a unique boundary between two highly mineralized tissues with very different matrix composition and physical properties. The nature of the boundary between the ectoderm-derived enamel and mesoderm-derived dentin is not known. This study was undertaken to identify the presence, type, and distribution of collagen at the dentin-enamel junction as an initial step in understanding its structural-functional role in dental occlusion. Sections of human teeth were demineralized with 0.1 M neutral EDTA and examined by high-resolution field-emission scanning electron microscopy at low accelerating voltage. Enamel and dentin were observed to be linked by many parallel 80-120-nm diameter fibrils, which were inserted directly into the enamel mineral and also merged with the interwoven fibrillar network of the dentin matrix. Immunogold labeling for collagen was visualized by secondary electron imaging and backscatter electron imaging at low accelerating voltage. The collagen fibrils at the junctional zone as well as in the dentin matrix were identified as Type I collagen. Collagenase digestion led to loss of the fibrillar structures and prevented immunogold labeling with antibody specific to Type I collagen. Consequently, the dentin-enamel junction can be regarded as a fibril reinforced bond which is mineralized to a moderate degree. PMID- 8429201 TI - An efficient method to detect calcitonin mRNA in normal and neoplastic rat C cells (medullary thyroid carcinoma) by in situ hybridization using a digoxigenin labeled synthetic oligodeoxyribonucleotide probe. AB - We report here an efficient and rapid method for the specific detection of calcitonin in tumor C-cells of medullary thyroid carcinoma (MTC). This occasionally aggressive tumor arises from the endocrine thyroid C-cells. Its principal marker is calcitonin, the predominant C-cell secretion, which is detected in patients and in our animal model by radioimmunoassay of the plasma, as well as by immunohistochemistry of thyroid tissues. Although calcitonin is easily detectable in normal C-cells, its content is greatly reduced in tumor cells owing to the disappearance of the secretory granules that store the mature peptide. This finding suggests cell dedifferentiation correlated with an increasing aggressivity of the tumor. We therefore developed a rapid detection of calcitonin mRNA by in situ hybridization on routine paraffin sections, using a synthetic oligodeoxyribonucleotide probe labeled with digoxigenin-dUTP. The reaction was detected with an anti-digoxigenin antibody conjugated with alkaline phosphatase, and the enzyme catalyzed the appearance of a dark blue color. The signal was exclusively restricted to the normal, hyperplastic, and tumor C-cells. It was specific, as increasing concentrations of the unlabeled oligonucleotide led to progressive disappearance of the reaction. Its sensitivity was slightly diminished as compared with corresponding frozen sections, but the intensity of the signal was quite acceptable. High levels of calcitonin mRNA were found in all normal and hyperplastic C-cells. They were increased in most of the tumor MTC cells, which did not correlate with the amount of intracellular peptide stores but explained the abnormally high basal levels of circulating calcitonin of the tumor-bearing rats. ISH is therefore of greater value than ICC for an early anatomopathological detection of this tumor. Our data show that the tumor cells are not "dedifferentiated." They only lack the granular compartment storing the mature peptide before exocytosis, but CT biosynthesis and the rest of the secretory process seem to be complete. Our results suggest that factors expressed in malignant C-cells affect basic cell mechanisms involved in the storage of the mature calcitonin, rather than the expression of the CALC gene. PMID- 8429202 TI - Postnatal development and sublobular distribution of cytochrome P-450 in rat liver: a microphotometric study. AB - To study the process of expression of cytochrome P-450 (P-450) in hepatocytes during development, we measured microphotometrically the P-450 content in periportal and perivenular hepatocytes of male rats during peri- and postnatal growth. From Day 19 of gestation to Day 5 after birth, P-450 content in both periportal and perivenular hepatocytes increased markedly (periportal 1046%; perivenular 819%). The content in periportal hepatocytes remained unchanged from 5 to 20 days of age, and increased slightly (24%) from 20 to 45 days of age. However, the content in perivenular hepatocytes increased progressively (105%) between 5 and 45 days of age. The difference in P-450 content became apparent between periportal and perivenular hepatocytes after 7 days of age. The content in periportal or perivenular hepatocytes reached the adult level at 45 days of age. Therefore, the perinatal period is the time at which a marked increase in P 450 occurs in hepatocytes throughout the liver lobule. The subsequent period before weaning is the time at which the sublobular heterogeneous distribution of P-450 appears. The period after weaning is the time at which a slight increase in P-450 content in periportal hepatocytes and a marked increase in the enzyme in perivenular hepatocytes takes place. PMID- 8429204 TI - Nutritional dependence of insulin-like growth factor (IGF) receptors in skeletal muscle: measurement by light microscopic autoradiography. AB - To determine the cellular location, capacity, and nutritional sensitivity of insulin-like growth factor (IGF) receptors, we measured the in vitro binding of [125I]-IGFs to skeletal muscle using light microscopic autoradiography. Muscle was collected from 8-month lambs that had received high or low nutrition diets (3% and 1.25% of body weight/day in pellets, respectively). Half of each group had also received growth hormone (0.25 mg/kg/day). Cryosections were incubated with [125I]-IGF alone or with unlabeled IGF-1, IGF-2, or insulin to characterize binding sites as probable Type 1 IGF, Type 2 IGF, or insulin receptors. [125I] IGF-1 was found to bind to blood vessels and Type 1 receptors in connective tissue (p < or = 0.001), but not to muscle fiber or nerves. In muscle from 6 month lambs that were fed or fasted, [125I]-IGF-1 bound to Type 1 receptors in connective tissue (p < or = 0.01 fed; p < or = 0.05 fasted) and muscle fiber (p < or = 0.05). The binding to connective tissue was also greater in fasted than in fed animals (p < or = 0.05). Binding of [125I]-IGF-2 to the Type 2 receptor was located in blood vessels and connective tissue (p < or = 0.01) and did not alter with fasting. Therefore, these experiments have demonstrated that Type 1 and Type 2 receptors vary in their distribution and nutritional sensitivity in skeletal muscle. PMID- 8429203 TI - Basement membrane proteoglycans in glomerular morphogenesis: chondroitin sulfate proteoglycan is temporally and spatially restricted during development. AB - We previously reported the presence of a basement membrane-specific chondroitin sulfate proteoglycan (BM-CSPG) in basement membranes of almost all adult tissues. However, an exception to this ubiquitous distribution was found in the kidney, where BM-CSPG was absent from the glomerular capillary basement membrane (GBM) but present in other basement membranes of the nephron, including collecting ducts, tubules, Bowman's capsule, and the glomerular mesangium. In light of this unique pattern of distribution and of the complex histoarchitectural reorganization occurring during nephrogenesis, the present study used light and electron microscopic immunohistochemistry to examine the distribution of BM-CSPG and basement membrane heparan sulfate proteoglycan (BM-HSPG) during prenatal and postnatal renal development in the rat. Our results show that the temporal and spatial pattern of expression of BM-CSPG during nephrogenesis is unlike that reported for other basement membrane components such as laminin, fibronectin, and BM-HSPG, all of which can be found in the earliest formed basement membranes of the vesicle-stage nephron. Although BM-CSPG is present in the basement membranes of the invading vasculature and ureteric buds, its first appearance in nephron basement membrane occurs during the late comma stage. In capillary loop-stage glomeruli of prenatal animals, BM-CSPG is present in the presumptive mesangial matrix but undetectable in the GBM. However, as postnatal glomerular maturation progresses BM-CSPG is also found in both the lamina rara interna and lamina densa of the GBM in progressively increasing amounts, being most evident in the GBM of 21-day-old animals. Micrographs of glomeruli from 42-day-old animals show that BM CSPG gradually disappears from the GBM and, by 56 days after birth, appears to be completely absent from the GBM, its pattern of distribution resembling that of the adult animal. Our results show that BM-CSPG is not required for the initial assembly of basement membranes but may in fact serve to stabilize basement membrane structure after histoarchitectural reorganization is completed. PMID- 8429205 TI - Seasonal changes in testicular structure and localization of a sperm surface glycoprotein during spermatogenesis in sea urchins. AB - Testes of the sea urchin, Strongylocentrotus purpuratus, undergo morphological changes consistent with a reproductive season that includes the winter months in the northern hemisphere (c. November to March). These changes can be observed in the basal germinal epithelia (BGE) of testicular acini as alterations in the gradients of meiosis and spermatogenesis from the peripheral (basal) to the inner (luminal) regions. Early in the season, large numbers of spermatogonia and primary spermatocytes along with large, spherical nutritive phagocytes (NPs) occupy most of the BGE. Relatively few secondary spermatocytes, spermatids, and spermatozoa are present. In the middle of the season more spermatocytes, spermatids, and spermatozoa are observed within well-defined spermatogenic columns that run between elongated NPs. Late in the season the BGE diminishes as the number of spermatogenic cells within it decreases dramatically and the lumen becomes distended with mature spermatozoa. To trace the origin and location of a 210 KD sperm surface antigen during spermatogenesis, a monoclonal antibody that recognizes this antigen, MAb J18/2, was used to probe testis sections. Indirect immunofluorescence microscopy detected the antigen throughout the BGE. Immunogold electron microscopy further localized it to the intracellular vesicles within spermatogenic cells of all stages. In mature spermatozoa the 210 KD surface antigen was detected not only on the sperm surfaces overlying the acrosomes and tails but also within the acrosome, suggesting the presence of an intracellular store of this antigen. Other than the occasional detection of antigen within the residual bodies of NPs, which contain phagocytosed spermatozoa, no antigen was detected in somatic cells of the testis, indicating that it is generated within the spermatogenic cells themselves. PMID- 8429206 TI - Coexistence of renin and cathepsin B in secretory granules of granular duct cells in male mouse submandibular gland. AB - Cathepsin B, a representative lysosomal cysteine proteinase, has been demonstrated to coexist with renin in secretary granules of rat pituitary LH/FSH cells and renal juxtaglomerular cells. We investigated immunocytochemically the localization of cathepsins B, H, and L in the submandibular gland of male mice, in which active renin is also produced. By light microscopy, granular immunodeposits for cathepsin B were detected in epithelial cells of the gland, particularly in granular duct cells and interstitial cells. Immunoreactivity for cathepsins H and L was mainly found in interstitial cells, although that for cathepsin H was weakly seen in acinar cells. By electron microscopy, immunogold particles indicating cathepsin B intensely labeled small granules near the Golgi complex of granular duct cells and weakly labeled large secretory granules, whereas those showing renin labeled both granules. Double immunostaining co localized immunogold particles showing renin and cathepsin B in small perinuclear granules near the Golgi complex. Some immunopositive granules seemed to be closely associated with the Golgi elements. These results indicate that the co localization of renin and cathepsin B is also seen in secretory granules of granular duct cells in the mouse submandibular gland, as seen in rat juxtaglomerular and LH/FSH cells. This suggests that cathepsin B is one of the possible candidates for the renin-processing enzyme. PMID- 8429207 TI - Improvements in the silver-staining technique for nucleolar organizer regions (AgNOR). AB - The silver-staining technique for nucleolar organizer regions (AgNOR) of Ploton et al., as popularized by Crocker, is being widely used for evaluation of nucleolar function, especially in neoplasia. It suffers from limited reliability, background staining, precipitates, and fading of the sections. Factors were identified that affect these problems. The oxidation-reduction level and gelatin used are particularly important. An improved procedure is presented which incorporates pre-reduction of the sections, selection of an optimal gelatin, and post-treatment of the sections to produce a permanent preparation. It is compatible with many fixatives and with other stains used before or after the silver stain. PMID- 8429208 TI - A method for exposing hidden antigenic sites in paraformaldehyde-fixed cultured cells, applied to initially unreactive antibodies. AB - We describe a simple protocol that allows the retrieval of masked or hidden intracellular antigens in cultured cells. The protocol is based on the exposure of paraformaldehyde-fixed and Triton X-100-permeabilized cells to guanidine hydrochloride (GdnHCl). We used it for localization of different antigens in BHK 21 cells by immunofluorescence microscopy. Our results showed that five out of six antibodies, initially unreactive, became excellent localization tools when used in conjunction with GdnHCl. Denaturation of fixed cells with GdnHCl did not affect the overall cell architecture, when monitored with different organelle specific antibodies. In two cases out of 17 the antigenic sites were lost after denaturation. However, this problem could be partially overcome by including low amounts of glutaraldehyde in the fixative. We think that this method could be generally useful in immunofluorescence localization studies, particularly in cases where the antibodies are known to react only with denatured antigens. PMID- 8429209 TI - PEG embedding for immunocytochemistry: application to the analysis of immunoreactivity loss during histological processing. AB - We have developed a protocol for the production and longterm storage of polyethylene glycol (PEG) sections for immunocytochemistry. Sections obtained by this protocol allow immunolabeling for many different antigens, such as intermediate filaments, macrophage markers, or neurotransmitter enzymes. Standard histological staining can also be performed on these sections. This fixation embedding system may therefore be of interest for histopathology of rare specimens, as well as for experimental research. Multiple labeling can be performed either on the same section or on consecutive thin sections, thus allowing a more thorough analysis of precious experimental material. We compare the advantages of PEG vs cryostat or vibratome sections. This protocol has been used to study the inactivation of antigenicity by paraffin embedding. We have identified the infiltration by paraffin as the antigenicity inactivating step, not dehydration or high temperature as generally thought. PMID- 8429210 TI - Large scale recovery of biologically active IgM (95% pure) from human plasma obtained by therapeutic plasmapheresis. AB - We have developed a large scale (10 g/batch) non-chromatographic method for the collection of IgM directly from plasma obtained from therapeutic plasmapheresis of patients. The technique uses sequential precipitations from ammonium sulfate and polyethylene glycol, and gives high yields of undenatured IgM antibodies. The biological activity of IgM prepared by this method has been demonstrated by systemically transfusing animals with the antibody and producing a demyelinating syndrome very similar to the antibody donor. Control IgM preparations exhibited no toxicity after daily injections of antibody for 8 weeks, showing that the method gives antibody with excellent bio-compatibility (Tatum, 1989). The method, which is effective for monoclonal and polyclonal antibodies, is useful when large amounts of antibodies are needed for passive transfer or other studies. PMID- 8429211 TI - Polymerase chain reaction in situ: intracellular amplification and detection of HIV-1 proviral DNA and other specific genes. AB - The ability to detect a single copy of a specific gene in situ has many advantages and multiple applications in molecular biology, pathology and cell biology. We report here a unique, highly sensitive and specific technique which can be utilized to detect a single copy of human immunodeficiency virus type I (HIV-1) provirus and other genes, at the single cell level, by in situ amplification of a portion of a gene sequence. In this method, a polymerase chain reaction (PCR) can be carried out in situ, in fixed cells, on specially designed glass slides. After amplification one can detect the amplified signals by the in situ hybridization method, utilizing either biotinylated probes or 32P-labelled probes. The early molecular events in the retroviral life-cycle of HIV-1, in specific target cells, are demonstrated utilizing in situ PCR. The techniques utilized in this procedure and various potential uses of this methodology are described. PMID- 8429212 TI - Responsiveness of the interleukin (IL)-6-dependent cell line B9 to IL-11. PMID- 8429213 TI - Improved sandwich enzyme-linked immunosorbent assays for transforming growth factor beta 1. AB - Transforming growth factor beta s (TGF-beta s), a highly homologous family of 25 kDa dimers, function as both paracrine and autocrine growth modulators which relay a broad spectrum of critical biological functions. Although sensitive bioassays for TGF-beta s have been described, their interference by other growth modulators have limited their application. We have previously described a highly sensitive (20-50 pg/ml) and specific sandwich enzyme-linked immunosorbent assay (SELISA) for TGF-beta 1, based on use of rabbit anti-TGF-beta 1 as the capture antibody. Widespread use of this assay was limited by the availability of this rather low-titer antibody. This report describes improved TGF-beta 1 SELISAs with comparable sensitivities to the above assay based on available sources of commercial monoclonal and polyclonal antibodies. These newly developed SELISAs are the most sensitive, specific, precise and convenient methods for the quantitation of TGF-beta 1 in complex biological fluids. PMID- 8429214 TI - Transplantation of CD4+ T cell clones into SCID mice. AB - Engraftment of congenic (BALB/c) or semi-syngeneic (dm2) CD4+ T cell clones into immunodeficient SCID mice was investigated in two experimental systems: the adoptive transfer of Iad-restricted, anti-host-reactive ('self-reactive') CD4+ T cell clones, and the adoptive transfer of OVA-specific CD4+ T cell clones. SCID mice transplanted with 10(4)-10(5) purified BALB/c CD4+ T cells select anti-host reactive T cells for engraftment. Anti-SCID-reactive T cell clones derived from such preselected CD4+ T cell populations could be successfully engrafted into secondary SCID recipients. In a second series of experiments, Iad-restricted, self-reactive BALB/c CD4+ T cell clones established in long-term culture from unselected, normal BALB/c spleen cell populations were transferred into SCID mice. Not a single clone from this panel could be transplanted into the immunodeficient host. Intravenous injection of OVA-primed dm2 (Ld-) CD4+ T cells into OVA-immunized young SCID (Ld+) mice repopulated the splenic and peritoneal T cell compartment of all transplanted mice. OVA-specific dm2 CD4+ T cells from the spleens of transplanted SCID mice were (sub)cloned under limiting dilution conditions in vitro. Only some of these clones repopulated OVA-immunized SCID recipients after retransplantation in vivo. Serial transfer experiments with a selected OVA-specific dm2 CD4+ T cell clone indicated that the route of cell transfer (i.p., but not i.v.), but not the route of OVA immunization of the host (i.v. or i.p.) were critical for successful engraftment of this clone. Hence, selected CD4+ T cell clones can be successfully engrafted into SCID mice if (i) the population from which clones are derived is preselected in SCID mice, (ii) the in vitro culture period of T cells is restricted to a few months, and (iii) alternative routes of cell transfer are tested. In infectious disease models, protective T cell clones can be identified in vivo using the described system. In addition, the importance of T cells with defined phenotype and effector functions in the pathogenesis of autoimmune disorders can be assessed in this system. PMID- 8429215 TI - In vitro immunization with antigen directly blotted from SDS-polyacrylamide gels to polyvinylidene difluoride membranes. AB - A new immunization method has been developed for the production of monoclonal antibodies. This technique uses small amounts of partially purified and weak immunogenic antigen, bound to membranes after blotting from SDS-PAGE. For this purpose two different membranes have been tested. Immobilon-P polyvinylidene difluoride (PVDF) membranes were less mitogenic than nitrocellulose membranes, and were therefore selected for the in vitro immunization using 6-pyruvoyl tetrahydropterin synthase as antigen. The in vitro immunization method was then used for the production of monoclonal antibodies against 6-pyruvoyl tetrahydropterin synthase, one of the key enzymes on the biosynthetic pathway of tetrahydrobiopterin, the natural cofactor of the mammalian aromatic amino acid hydroxylases. The antibodies obtained were mainly of the IgM type. PMID- 8429216 TI - Optimisation of small-scale coupling of A5B7 monoclonal antibody to carboxypeptidase G2. AB - Conjugates of F(ab')2 fragment of the monoclonal antibody A5B7 coupled to carboxypeptidase G2 (CPG2) have been produced using the heterobifunctional reagents 2-mercapto-[S-acetyl]acetic acid, N-hydroxysuccinimide ester (SATA) and m-maleimidobenzoyl-N-hydroxysuccinimide ester (SMPB). The effect of various levels of modifying reagent on enzyme activity and antigen binding activity were determined, and it was shown that whilst CPG2 is relatively sensitive to modification, insertion of three maleimide groups per CPG2 resulted in the loss of 30% of enzyme activity; A5B7 F(ab')2 was insensitive to modification, little or no activity being lost. The coupling efficiency of the reaction was shown to be fairly constant over a wide range of substitution levels. There was thus no advantage to be gained in using high substitution levels, which may result in loss of enzyme activity. The formation of undesired high molecular weight aggregates could be controlled by adjustment of the protein concentration during the final coupling step. PMID- 8429217 TI - The use of palmitate-conjugated protein A for coating cells with artificial receptors which facilitate intercellular interactions. AB - A method is described in which palmitate-conjugated protein A (pal-protein A) incorporated onto cell membranes is used to anchor antibodies on the surface of cells. Pal-protein A was stabilized on the cell surfaces via insertion of the hydrophobic palmitate moieties into the phospholipid bilayer of the plasma membrane. This membrane-associated pal-protein A retained its binding affinity for the Fc region of rabbit immunoglobulin G (IgG) molecules. Using this system, cells can be coated with antibody (Ab) molecules of a desired specificity that can serve as artificial receptors for soluble or cell surface antigens (Ags). In this report, we show that A22.E10 T hybridoma cells coated with rabbit anti-mouse IgG (RaMIgG) exhibited increased adhesiveness to surface IgG positive mouse B cells. A high level of heterotypic intercellular conjugation was observed in cells coated with specific artificial receptors as compared to cells coated with nonspecific artificial receptors. PMID- 8429218 TI - Investigation of human platelet alloantigens and glycoproteins using non radioactive immunoprecipitation. AB - Sensitive techniques to detect platelet antibodies are needed for the investigation of immune thrombocytopenic syndromes such as neonatal alloimmune thrombocytopenia and post-transfusion purpura. Radioimmunoprecipitation has proved useful in the investigation of platelet-antibody interactions; however, the requirement for a radioactive label is a disadvantage. We describe the immunoprecipitation of human platelet proteins labelled with nonradioactive NHSS biotin and compare the results with proteins labelled with 125I. The efficiency of labelling was evaluated by immunoprecipitation using well-characterized human anti-platelet antisera and murine monoclonal antibodies. The immunoprecipitated proteins were separated by SDS-PAGE, transferred to nitrocellulose and detected using streptavidin-horseradish peroxidase and a chemiluminescent substrate with exposure to X ray film. The biotinylation technique labelled glycoproteins Ia/IIa, Ib/IX, IIb/IIIa, IV, and p175 which carry all of the known platelet alloantigens and isoantigens. It was as sensitive as radiolabelling and had the advantage of labelling GPs Ib beta and IX, which were not labelled using radioiodine. Human sera containing alloantibodies to HPA-1a on GP IIIa, HPA-3a on GP IIb, HPA-5a and HPA-5b on GP Ia, Govb on p175, and the isoantibody Naka on GP IV precipitated the corresponding biotinylated proteins. Biotinylated proteins could be detected using a 30 s exposure compared to 2 days or longer for 125I. Immunoprecipitation of human platelet glycoproteins labeled with NHSS-biotin is a fast and sensitive alternative to conventional radioimmunoprecipitation for the study of human platelet antigens. PMID- 8429219 TI - Analysis of peripheral blood mononuclear cells in rodents by three-colour flow cytometry using a small-volume lysed whole blood technique. AB - Techniques were adapted to permit phenotypic analysis of peripheral blood leucocytes in rats and mice using small volumes of lysed whole blood and single, dual- or triple-colour immunofluorescence staining with flow cytometry. Red cell lysis using proprietary lysing solutions provides populations of lymphocytes, monocytes and granulocytes clearly differentiable by their light scatter properties. In the present study, measurement of functional and activation T lymphocyte subsets was performed by single- and dual-colour flow cytometry in both species and in excess of 5000 lymphocytes could be counted using as little as 10 microliters of whole blood. Triple-colour fluorescence, using antibodies labelled with fluorescein, phycoerythrin and biotin permitted simultaneous detection of three phenotypic markers. This novel technique has two major advantages, maximising the phenotypic information which may be obtained and using volumes of blood which are sufficiently small that they can be drawn regularly from the same animal. PMID- 8429220 TI - Two-colour combination enzyme-linked immunosorbent assay for the simultaneous detection of HBV and HIV infection. AB - To reduce the cost, time and waste in screening for HIV and HBV infections a combined assay for HIV-1 and -2 antibodies and HBsAg has been developed. Monoclonal anti-HBs antibodies were co-immobilized with synthetic peptides representing immunodominant regions of HIV-1 and -2. The presence of anti-HIV antibodies in the samples was detected with alkaline phosphatase-labelled anti human IgG and of HBsAg with horseradish peroxidase-labelled monoclonal anti-HBs antibodies by a sequential substrate reaction. In this assay, HBsAg was detectable in a concentration range between 0.25 and 0.30 U/ml and the results were available within 3 h. The specificity, tested on 5000 serum samples from blood donors after confirmation, was 99.8% for HBsAg and 99.5% for anti-HIV antibody detection. All serum samples taken from 600 HIV-1- and 115 HIV-2 infected individuals were correctly classified as reactive. The two-colour HBsAg anti-HIV-1/-2 combination ELISA meets all the requirements of single parameter assays with regard to precision, stability and robustness. PMID- 8429221 TI - Pitfalls in the use of ELISA to screen for monoclonal antibodies raised against small peptides. AB - Small peptides are often conjugated to large carrier proteins such as thyroglobulin to increase their immunogenicity. Antibodies are raised against peptide, thyroglobulin and contaminants, such as immunoglobulins or BSA in the thyroglobulin preparation, but anti-contaminant antibodies will not be revealed if immune serum is diluted in buffer containing immunoglobulin-contaminated BSA. This may lead to the development of an inappropriate hybridoma-screening assay which detects more anti-contaminant than anti-peptide antibodies if ELISA plates are coated with a capture antibody or blocked with BSA. Dilution of culture supernatants in buffer containing BSA and Tween 20 minimises the risk of false positive results and makes plate-blocking unnecessary. The high affinity peptide specific antibodies in immune serum, which are more readily detected than lower affinity monoclonal antibodies, may result in an inadequately sensitive hybridoma screening ELISA. PMID- 8429222 TI - Rapid induction of anti-idiotypic responses to unmodified monoclonal antibodies from syngeneic mice following primary immunization. AB - A single foot-pad immunization in adjuvant of BALB/c mice with non-modified BALB/c monoclonal antibodies (HyHEL 5, 9 and 10) specific for hen egg lysozyme permitted isolation of anti-idiotypic monoclonal antibodies 10 days later. An evaluation of different screening tests revealed that antibodies were detected more easily by isotype-specific or direct binding assays than by cross-linking ELISA procedures. These results were confirmed by a direct cell-binding assay on B cells transgenic for one of the immunizing antibodies. The use of these cells also permitted an evaluation of the ability of these antibodies to inhibit antigen binding under conditions in which the target antibody, in its cell surface configuration, is minimally modified by potential artifacts induced by purification or fixation to a solid support. This study demonstrates that anti idiotypic responses to anti-protein antibodies may be rapidly generated in syngeneic animals. PMID- 8429223 TI - Avidin attachment to red blood cells via a phospholipid derivative of biotin provides complement-resistant immunoerythrocytes. AB - Preincubation of red blood cells (RBC) in an aqueous dispersion of biotin phosphatidylethanolamine (biotin-PE) provides binding sites for avidin on the surface of these cells (up to 5 x 10(5) avidin molecules per cell). Previously we have shown that biotin covalently attached to the surface of RBC by a chemical reaction with biotin N-hydroxysuccinimide ester permits attachment of avidin to these cells, resulting in the activation of the alternative pathway of complement with subsequent cell lysis. However, avidin attached to RBC via biotin-PE did not cause complement activation. This is not due to the stabilizing action of biotin PE. In contrast, various phospholipids, including biotin-PE, enhance the lysis of RBC induced by hemolytic antibodies via the classical complement pathway. The potential of avidin-coated RBC to act as activators of the complement alternative pathway depends on the method of biotin attachment to RBC. Complement-resistant avidin-coated RBC can specifically bind biotinylated antibodies. These immunoerythrocytes effectively and specifically bind to the antigen-coated surface and are not lysed by complement even in the presence of soluble antigen. These data extend the possible applications of immunoerythrocytes in drug targeting. PMID- 8429224 TI - An immunoenzyme technique for the identification of granulocyte-macrophage colony stimulating factor (GM-CSF) receptors using digoxigenated-GM-CSF. AB - A method for detecting granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors has been devised using human macrophages and a GM-CSF/IL-3-dependent human megakaryoblastic leukemia cell line (M-07e). Recognition of the factor binding site was accomplished by linking recombinant human (rh) unglycosylated GM CSF previously labeled with digoxigenated compounds. Digoxigenates were able to link amino and sulphydryl groups of the soluble factor and an immunoperoxidase technique using monoclonal anti-digoxigenin antibody was employed to demonstrate the interaction. To support morphological data cross-linking analysis was performed with M-07e cells using digoxigenated-rh-GM-CSF. Macrophages and M-07e cells incubated with digoxigenated-rh-GM-CSF showed intense positivity by the immunoperoxidase technique. In cross-linking, M07e cells showed a 96 kDa band corresponding to receptor plus bound factor. This technique permits a high degree of specificity in the detection of GM-CSF receptors with good morphological preservation of cellular detail. PMID- 8429225 TI - A rapid and sensitive immunoassay for antibodies against alloantigens on human platelet glycoproteins (BIPA). AB - A rapid and sensitive enzyme immunoassay has been developed for the detection and specification of alloantibodies against human platelet antigens (HPA) on platelet membrane glycoproteins. Human platelet glycoprotein complexes were coupled by monoclonal antibodies to immunomagnetic beads with sheep anti-mouse IgG directed antibodies. These beads served as solid-phase target antigens for human alloantibodies in a modified MAIPA assay. This test provided high sensitivity, permitting the identification of antibodies against membrane receptors and required only 1 h of assay time. The specificity and sensitivity of this assay were evaluated in serum samples from polytransfused patients and compared with standard immunoassays employing antigen coated immunowells. PMID- 8429226 TI - Elimination of extracellular bacteria by antibiotics in quantitative assays of bacterial ingestion and killing by phagocytes. AB - In vitro assays to quantify the bactericidal capacity of phagocytes require the removal of extracellular bacteria from the reaction mixture before intracellular bacteria are released from the phagocytic cells and counted. This may be achieved by using an antibiotic, such as gentamicin, which has a rapid bactericidal action coupled with limited ability to cross cytoplasmic membranes. In this study, we investigated the susceptibility of Escherichia coli and Yersinia enterocolitica to killing by mouse peritoneal macrophages. Extracellular bacteria in the reaction mixture were eliminated with gentamicin, whereafter the antibiotic was enzymatically inactivated using gentamicin acetyl transferase (GAT). The method has few washing stages, thus reducing the likelihood of inadvertent removal of macrophages or bacteria from the reaction mixture. The assay disclosed clear differences in the ability of E. coli and Y. enterocolitica to survive phagocytosis by macrophages. PMID- 8429227 TI - Flow cytometric analysis of surface major histocompatibility complex class II expression on human epithelial cells prepared from small intestinal biopsies. AB - A technique for preparing viable, single cell suspensions of the epithelial layer of small intestinal tissue obtained endoscopically is described. Constant agitation of four biopsies for 60 min in the presence of chelating and reducing agents gave yields of 1.2-6.7 x 10(6) cells, of which 11-30% were intraepithelial lymphocytes (IEL). Passage through a nylon wool column removed dead cells. This preparation was suitable for flow cytometric analysis. Using this technique, surface MHC class II molecule expression was studied in 14 patients with normal small intestinal mucosa. Fluorescence labelling of these cells showed strong HLA DR expression by epithelial cells (EC), DP was expressed less strongly, while little DQ expression could be detected. This technique demonstrates that small intestinal biopsies taken during routine endoscopy can yield adequate numbers of viable epithelial cells to perform flow cytometric analysis. PMID- 8429228 TI - Quantitative analysis of lymphokine mRNA expression by a nonradioactive method using PCR and anion exchange chromatography. AB - Amplification of DNA by the polymerase chain reaction (PCR) has become an efficient tool in the study of gene expression. We describe the use of HPLC anion exchange chromatography to quantitate PCR products amplified from cDNA. The technique circumvents the use of both radioactivity and gel electrophoresis. We show that the method permits accurate quantitation of the gene product of interest and provides a clear separation of specific and non-specific products. The technique was applied to quantitate TNF-beta mRNA levels in unstimulated and stimulated mouse T cells. PMID- 8429229 TI - Rapid determination of hepatitis B surface antigen (HBsAg) by latex agglutination using an integrating sphere turbidimetric assay. AB - A method for determining levels of serum HBs antigen has been developed, applying the principles of the integrating sphere turbidimetric assay (ISTA). Using this method, the minimum detectable level of HBs antigen is 15 ng/ml, i.e., it is three times more sensitive than the reversed passive hemagglutination (RPHA) method. Reproducibility and specificity are also excellent with ISTA. With a cut off level of 20 ng/ml, the highest reading possible with this method is 1000 ng/ml. Serum HBs antigen can readily be measured by this method if a fully automated EL-1000 analyzer is used. This rapid and simple method of measurement should be clinically useful. PMID- 8429230 TI - Detection of lymphocyte Fc gamma receptor-blocking factors by the EA rosette inhibition assay. Refinement of the conventional method and development of a novel flow-cytometric assay. AB - Serum factors which interact with human peripheral blood lymphocyte Fc gamma receptors (Fc gamma Rs) may be detected in vitro by the EA rosette inhibition assay (EARIA). This assay has been used to detect circulating immune complexes and certain alloantibodies directed against cell surface antigens situated in close proximity to Fc gamma Rs. Three main types of FcR-blocking factor have been demonstrated by the EARIA in human serum following exposure to alloantigens. A strong correlation was observed between the presence of one of these FcR-blocking factors (FcBF1) and human renal allograft survival. This factor was previously shown to bind preferentially to CD32+ B cells and to inhibit antibody synthesis. In this study we have shown that detection of FcBF1 by the EARIA depends on the type of erythrocyte and on the amount of antibody used to sensitise the erythrocytes. Furthermore, we have developed a flow-cytometric version of the EARIA which is rapid, reproducible and, most importantly, objective. Inter laboratory comparisons using this standardised EARIA should now be possible. PMID- 8429231 TI - Effects of adequate versus inadequate treatment of cutaneous manifestations of Lyme borreliosis on the incidence of late complications and late serologic status. AB - Eighty-two patients who were treated at the Department of Dermatology, Innsbruck, Austria, from 1980 to 1987 for cutaneous manifestations of Lyme disease were subjected to a clinical follow-up investigation aimed at detecting dermatologic, neurologic, and internal late complications of borreliosis. Only 54 of these patients had received adequate antibiotic treatment according to current standards. Also, their sera were investigated for the presence of immunoglobulin G (IgG) and IgM Borrelia burgdorferi antibodies by an indirect immunofluorescence assay, three different enzyme-linked immunosorbent assays, and immunoblotting. As a control, the sera of 126 healthy blood donors were investigated with the same assays. Results showed no unambiguous clinical late complications of Lyme borreliosis, even in inadequately treated or untreated patients. Seropositivity varied considerably according to the assay used; the indirect immunofluorescence assay yielded the highest scores. The proportion of seropositive results (immunofluorescence assay) was 59% in patients with erythema chronicum migrans, 69% in those with lymphocytoma cutis, and 100% in those with acrodermatitis chronica atrophicans (overall 63%); in contrast, only 31% of the blood donor control group were found to be seropositive. Seropositivity did not correlate with adequacy of treatment, interval between onset of symptoms and treatment, time span since treatment, age of patients, and presence of antinuclear antibodies. Immunoblot pattern showed high incidence of antibodies against the 29/31-kD (outer surface proteins OspA and OspB) and 55/58-kD antigens in general and against the 41-kD protein (flagellin) in patients with acrodermatitis chronica atrophicans only. PMID- 8429232 TI - Lysosome-associated membrane protein-1 (LAMP-1) is the melanocyte vesicular membrane glycoprotein band II. AB - Coated vesicles play a critical role in the process of melanogenesis. Antisera raised against a coated vesicle fraction from mouse melanoma cells recognize two major glycoprotein antigens, band I (47-55 kd) and band II (90-120 kd). We demonstrate that band II is lysosome-associated membrane protein 1 (LAMP-1) by the following criteria: 1) the molecular weight and abundance of LAMP-1 varies among tissues but is always identical to that of band II; 2) band II and LAMP-1 co-migrate in sucrose gradient sedimentation studies; 3) immunodepletion of cell extracts with antivesicle serum removes all LAMP-1; and 4) intact organelles immunoisolated with antivesicle serum contain band II and LAMP-1. Our results further confirm the long-suspected relationship between melanosomes and the lysosomal lineage of organelles. PMID- 8429233 TI - Migration of human melanoma cells on hyaluronate is related to CD44 expression. AB - Phenotypic and functional aspects of melanoma-hyaluronate interactions were investigated by studying the expression of CD44, cell migration, and transmembrane penetration of human melanoma cell lines on hyaluronate-coated substrates. Expression of CD44 was tested by flow cytometry on seven human melanoma cell lines. Strong reactivity with anti-CD44 monoclonal antibody was observed in four of seven of the cell lines. Migration studies of CD44(+) cell lines on hyaluronic acid- and chondroitin-6-sulfate-coated substrates, using time lapse video-microscopy, showed a dramatic dose-dependent increase in migration rate on hyaluronate but not on chondroitin-6-sulfate. Moreover, CD44(-) cell lines showed no modification in migration rate on either substrate. Addition of soluble hyaluronate produced a dose-dependent inhibition of acceleration of CD44(+)cells on hyaluronate-coated substrates, whereas addition of chondroitin-6 sulfate had no effect. Migration inhibition experiments with soluble CD44 (CD44 receptor globulin) also showed specific blocking of the migration of CD44(+) cells on hyaluronate. Haptotactic invasion was increased in CD44(+) cell lines through hyaluronate-coated polycarbonate membranes, whereas no change was detected on chondroitin-6-sulfate-coated membranes. CD44(-) cell lines showed no response to either type of coating. In the melanoma cell lines tested, the expression of CD44 correlated with in vitro migration and invasiveness on hyaluronate substrates. Taken together, our data are consistent with the suggestion that CD44 may play a role in stimulating in vivo aggressiveness of tumors through hyaluronate-rich stroma. PMID- 8429234 TI - Cytochemical localization of hyaluronan in rat and human skin mast cell granules. AB - Rat and human skin were processed either by osmium tetroxide/microwave fixation followed by embedding in epoxy resin or by glutaraldehyde/microwave fixation and low-temperature embedding in Lowicryl K4M. Hyaluronan-binding proteins and link proteins (LP) were isolated from bovine nasal cartilage, coupled to 15-20-nm gold particles and employed as markers in a one-step post-embedding procedure for identifying hyaluronan (hyaluronic acid) at the ultrastructural level. Mast cell granules of both species were labeled. The specificity of the hyaluronan-binding probes was demonstrated by treatment of sections with testicular hyaluronidase, Streptomyces hyaluronidase, and chondroitinase ABC, and pre-incubation of probes with hyaluronan oligosaccharides. The results suggest that mast cell granules are a rich source of hyaluronan; this finding may account for the striking concurrence of hyaluronan accumulation with a mastocytotic condition in many tissues undergoing pathologic changes. PMID- 8429235 TI - Pigment production in murine melanoma cells is regulated by tyrosinase, tyrosinase-related protein 1 (TRP1), DOPAchrome tautomerase (TRP2), and a melanogenic inhibitor. AB - Using antibodies that recognize either tyrosinase, tyrosinase-related protein-1 (TRP1), or tyrosinase-related protein-2 (TRP2, DOPAchrome tautomerase), the quantities of those melanogenic enzymes were analyzed in five melanoma cell lines that possess various degrees of melanin production. All cells except JB/MS-W increased melanin production four to 30 times after 4 d of melanocyte-stimulating hormone (MSH) treatment. Melanin production by JB/MS-W cells was always under background, with or without MSH treatment. There was a positive correlation between quantities and synthetic rates of those melanogenic enzymes and their melanin formation or DOPAchrome tautomerase activities. The activity of a heat resistant melanogenic inhibitory factor was also analyzed. The results showed, surprisingly, that pigmented cells showed higher levels of melanogenic inhibitors activity. Tyrosinase activity was increased dramatically whereas the level of melanogenic inhibitor was remarkably decreased following MSH treatment. Interestingly, melanogenic inhibitor derived from JB/MS-W cells suppressed not only tyrosinase but also DOPAchrome tautomerase, another enzyme functional in melanin production. These results clearly suggest that melanin production is regulated by a subtle balance between the activities of these enzymes and other factors such as the melanogenic inhibitor. PMID- 8429236 TI - Eosinophil migration in atopic dermatitis. I: Increased migratory responses to N formyl-methionyl-leucyl-phenylalanine, neutrophil-activating factor, platelet activating factor, and platelet factor 4. AB - Eosinophil granular protein deposits have been demonstrated in lesional atopic dermatitis skin. This suggests active tissue infiltration of eosinophils. To find an explanation for the tissue influx of eosinophils, eosinophil migration was studied in vitro by means of a microchemotaxis assay. Eosinophils from the circulation of patients with atopic dermatitis showed an altered capacity to respond to chemotactic stimuli in vitro compared with eosinophils from healthy donors. Eosinophils from patients with atopic dermatitis had significantly increased migratory responses toward dose ranges of N-formyl-methionyl-leucyl phenylalanine, neutrophil-activating factor, platelet-activating factor, and platelet factor 4. Eosinophils from normal individuals did not respond to N formyl-methionyl-leucyl-phenylalanine and neutrophil-activating factor and responded only slightly to platelet factor 4. The migratory responses toward tumor necrosis factor-alpha and complement factor C5a were identical in both groups. Interleukin-5, an eosinophil-selective cytokine, is a strong modulator of the migratory responses to these chemotaxins in eosinophils from normal donors. A migratory response toward N-formyl-methionyl-leucyl-phenylalanine and neutrophil activating factor was induced by interleukin-5, whereas the migratory response toward platelet-activating factor and platelet factor 4 was markedly potentiated. In contrast, the response to complement fragment C5a was only slightly influenced. Our findings indicate that the increased migratory responsiveness of eosinophils from patients with atopic dermatitis to various chemotaxins reflects in vivo "priming" of eosinophils, presumably by circulating cytokines such as interleukin-5. This in vivo "priming" is not optimal because it can be further potentiated by renewed contact with interleukin-5. PMID- 8429237 TI - Up-regulation of alpha 4 integrin on activated Langerhans cells: analysis of adhesion molecules on Langerhans cells relating to their migration from skin to draining lymph nodes. AB - After hapten application, epidermal Langerhans cells migrate into the regional lymph nodes through dermal lymphatics. Recently, we have demonstrated that some of them take the phenotypic and functional characteristics similar to those of in vitro cultured Langerhans cells, before disappearing from the epidermis. To analyze the mechanisms underlying the migration of Langerhans cells, we studied the expression of several adhesion molecules on freshly isolated LC and cultured LC. Pgp-1 (CD44), intercellular adhesion molecule 1, and alpha 4 integrin were strongly expressed on cultured Langerhans cells. Among them, only alpha 4 integrin was strongly up-regulated by cultured Langerhans cells, because its expression by freshly isolated Langerhans cells was very weak. This up-regulation of alpha 4 integrin was also observed on in vivo activated Langerhans cells in the epidermis and draining lymph nodes after hapten application. These data suggest a possible role played by VLA-4 in the migration of Langerhans cells from the epidermis into the regional lymph nodes after hapten application. PMID- 8429238 TI - Localization of 92-kDa type IV collagenase in human skin tumors: comparison with normal human fetal and adult skin. AB - To investigate the role of secreted metalloproteinases in the behavior of skin tumors we have studied immunoreactivity for 92-kDa type IV collagenase (92T4Cl) in benign tumors of sweat glands, basal cell carcinomas (BCC), baso-squamous cell carcinomas (BSCC), and squamous cell carcinomas (SCC). In all tumors, the enzyme was found in stromal cells, but not in tumor epithelium. 92T4Cl-positive cells contained the common leukocyte antigen HLe-1(CD45) and the polymorphonuclear leukocyte-specific antigen PMN-8C7. Only a few 92T4Cl-positive cells expressed either macrophage-specific Leu-M5 or eosinophil-specific cationic protein antigens. In benign sweat gland tumors, and in the majority of nodulocystic and adenoid BCCs, 92T4Cl-positive cells were relatively rare and no extracellular deposition of the enzyme was found. In the more aggressive tumors examined, SCCs, BSCC, recurrent, infiltrative, and morpheaform BCCs, 92T4Cl-positive cells were very abundant. In addition, a significant quantity of extracellular enzyme was deposited both within the extracellular matrix adjacent to the tumor nests and in their basement membrane zone. In normal adult skin only a few scattered 92T4Cl containing cells were found in the dermis whereas in fetal skin, groups of 92T4Cl positive, HLe-1-negative cells were present in the upper dermis. These observations suggest that in cutaneous tumors, extensive infiltration of 92T4Cl containing polymorphonuclear leukocytes and the extracellular deposition of the enzyme in the basement membrane zone are signs of more aggressive tumor behavior. PMID- 8429239 TI - Detection of the 170-kDa bullous pemphigoid antigen by immunoprecipitation. AB - There has been controversy concerning the nature of the bullous pemphigoid (BP) antigen: immunoprecipitation identified BP antigen as a single, unique 230-kDa protein, whereas immunoblot analysis showed multiple antigen molecules, mainly 230- and 170-kDa proteins. In this study, to further characterize the 170-kDa protein, we have examined whether the 170-kDa protein is detected by immunoprecipitation. Extracts of human squamous cell carcinoma cells revealed the 170-kDa protein with immunoblot analysis. Although the conventional immunoprecipitation detected only the 230-kDa protein, some BP sera that detected the 170-kDa protein with immunoblotting also precipitated the 170-kDa protein with our modified immunoprecipitation, in which the cells were extracted with 1% sodium dodecylsulfate (SDS) buffer and reacted with the sera under reduced SDS concentration. The 170-kDa protein-specific BP sera clearly showed hemidesmosomal plaque staining with immunofluorescence of cultured cells. These results indicate that the 170-kDa protein is indeed one of the BP antigens and that the 230- and 170-kDa BP antigens are integrated in different ways in hemidesmosomes. PMID- 8429240 TI - Adherens junctions: demonstration in human epidermis. AB - Adherens junctions are intercellular and cell-matrix junctions that, like desmosomes and hemidesmosomes, mediate adhesion of cells to each other or to matrix structures. These junctions have been detected recently in cultured human keratinocytes, indicating that they may be of importance in epidermis. To investigate the localization of adherens junctions in normal epidermis, we examined human epidermis, human oral mucosa, and monkey esophagus for the presence of vinculin, a major protein of the intracellular plaques of adherens junctions that is thought to be present in all adherens junctions. Western blot analysis demonstrated vinculin in extracts of epidermis. Immunohistochemistry of vinculin in these tissues displayed two distinct locations for adherens junctions: i) at the dermal-epidermal junction, and ii) in the region of cell cell contacts in all layers of the epidermis. The location of vinculin in the region of the epidermal-dermal junction is reminiscent of the distribution of vinculin-containing focal contacts in cultured keratinocytes, and the intercellular staining of vinculin in epidermis is consistent with the presence of vinculin in adherens junctions in cultured keratinocytes at sites of cell-cell contact. These results demonstrate that adherens junctions are present in human epidermis, oral mucosa, and monkey esophagus. Vinculin-containing junctions in epidermis may be important in the pathogenesis of skin diseases involving alterations in intercellular integrity. PMID- 8429241 TI - Effects of systemic indomethacin, meclizine, and BW755C on chronic ultraviolet B induced effects in hairless mouse skin. AB - Chronic exposure of hairless mice to ultraviolet B (UVB) radiation is associated with inflammation as well as an altered macromolecular composition of the dermis. This study was designed to determine whether or not various systemic anti inflammatory agents inhibit chronic UVB-induced changes in the macromolecular content of the dermis and, if so, whether each agent had the same or different effects. The agents and doses were chosen for their ability to inhibit the changes induced by a single exposure to UVB radiation (increased vasopermeability, neutrophil accumulation, and skin-fold thickness). Indomethacin, a cyclooxygenase inhibitor, and meclizine, an H1 histamine receptor antagonist, were administered from slow-release pellets. BW755C, a combined cyclooxygenase and lipoxygenase inhibitor, was administered intraperitoneally 30 min prior to UVB exposure. Animals were exposed to UVB three times per week for 20-26 weeks or were unirradiated. The elastin, glycosaminoglycan and collagen content of the skin were determined by measuring the desmosine, uronic acid, and hydroxyproline levels, respectively. The amount of each macromolecule per area of skin increased after chronic UVB exposure. The increase in desmosine was inhibited by indomethacin; the increase in hydroxyproline was inhibited by meclizine and BW755C. None of the agents inhibited the uronic acid increase. These results suggest that chronic inflammation contributes to the dermal changes seen in chronically UVB-exposed skin and that different inflammatory mediators are involved in the increases observed in elastin, glycosaminoglycans, and collagen. PMID- 8429242 TI - UVA-induced ultrastructural changes in hairless mouse skin: a comparison to UVB induced damage. AB - In this ultrastructural study, albino hairless mice were irradiated with long wavelength ultraviolet (UVA) (340-400 nm) thrice weekly for 32 weeks for a cumulative dose of 8000 J/cm2. Biopsies were taken from these mice, from age matched unirradiated controls, and from mice irradiated with UVB for 20-30 weeks with a cumulative dose of approximately 6-9 J/cm2. The most striking UVA-induced changes were 1) elastic fiber hyperplasia without evidence of fiber disintegration, 2) a large increase in randomly deposited microfibrils; 3) massive duplication of vascular basement membrane; 4) extensive endothelial cell damage; and 5) collagen fibers with smaller diameters but without apparent damage. By contrast, after UVB, the hyperplastic elastic fibers frequently appeared to be degraded. Microfibrils were only moderately increased and remained in an organized array. Also, unlike with UVA, the epidermal basement membrane was duplicated whereas that of the vessels was mainly spared. Collagen fibers showed evidence of dissolution. Thus, ultrastructural features provide further evidence that skin damage induced by UVA can be dissimilar to that induced by UVB. PMID- 8429243 TI - Effect of age and diet on stratum corneum barrier function in the Fischer 344 female rat. AB - Stratum corneum barrier function in the female Fischer 344 rat was assessed in animals ranging from 11 to 144 weeks of age, which were either fed ad libitum or a caloric-restricted diet at 60% ad libitum. Caloric restriction has been shown to increase the life span of rodents and delay age-associated degenerative diseases. The penetration of water, lidocaine, and hydrocortisone were measured in vitro using the finite-dose technique in Franz diffusion chambers. Water permeability did not significantly change with either age or diet condition (with the single exception of the ad libitum 144-week animals). However, between 11 and 44 weeks, both lidocaine and hydrocortisone permeability increased with age in both diet groups. By 44 weeks the caloric restriction animals had significantly greater permeability to lidocaine and hydrocortisone then their ad libitum age cohorts. Between 44 and 144 weeks, skin permeability in the ad libitum animals continued to increase with age, whereas permeability in the caloric restriction animals declined. By 144 weeks the caloric restriction skin was less permeable than the age-matched ad libitum animals but still more permeable than the 11-week animals. The data show that the barrier properties of stratum corneum change with age in the female rat and that this change can be modulated by caloric restriction. PMID- 8429244 TI - [Effect of pulsed ultrasound exposure on development of early embryos]. AB - Ultrasonography is indispensable for medical treatment, particularly in the field of reproductive medicine. Therefore, it is important to confirm safety when exposing preimplantation embryos to ultrasound. For this purpose, the study was designed to elucidate the bioeffects of pulsed ultrasound on preimplanted mice embryos. One-cell stage embryo, 2-cell stage embryo, morula and expanded blastocysts of mice in vitro cultured were exposed to experimental pulsed ultrasound for 15 minutes in degassed water at 37 degrees C. The ultrasound used in this study was 2MHz; [spatial-average temporal-average intensity (ISATA): 120mW/cm2, maximum intensity (Im): 39.1W/cm2, the peak rarefactional pressure (Pr): 1.0MPa, pulse width: 10 mus' and pulse repetition frequency: 1.0kHz.]. The development to expanded blastocyst and the nucleic acid synthesis in expanded blastocyst were compared in the embryos exposed (study group) and those not exposed (control) to the ultrasound. There were no significant differences between the two groups in the rates of development to expanded blastocysts and the nucleic acid synthesis. The temperature of the irradiated medium in cell container straw did not rise. These results suggest that pulsed ultrasound at the intensity used in this study does not have a harmful effect on early preimplantation embryos. PMID- 8429245 TI - [Mutational activation of H-ras and K-ras (Codon 12,61) genes in 7,12 dimethylbenz(a)anthracene induced rat ovarian tumors]. AB - We had studied the histogenesis and p21 expression in ovarian tumor induced by 7,12-dimethylbenz(a)anthracene (DMBA). The present study was done to elucidate point mutation of the ras gene in tumors. Cases with immunohistological p21 expression; 3 adenomas, 5 adenocarcinomas and 2 sarcomas were examined by polymerase chain reaction (PCR). 1) The products of PCR were confirmed the presence of amplified sequences by electrophoresis. 2) No point mutation of H-ras (Codon 12,61) or K-ras (Codon 12,61) genes was found in the ovarian tumors induced by DMBA. 3) These results suggested that the mutational activation of ras genes in tumorigenesis was dependent on another mechanism. PMID- 8429246 TI - [Etoposide/cis-platinum (CDDP) combined chemotherapy for ovarian cancer: evaluation of optimum schedule for CDDP administration in chronic continuous exposure of ovarian cancer cells to low-dose etoposide in vitro]. AB - Combined chemotherapy with etoposide and cis-platinum (CDDP) is considered a second line regimen for refractory cases of ovarian cancer. In addition to the time-dependent cytocidal kinetic of etoposide, much attention has been paid to low-dose continuous administration of etoposide. In this study, ovarian cancer cells (SHIN-3) were continuously exposed to low-dose etoposide in vitro to determine the optimum schedule for CDDP administration. Etoposide concentrations of 1-3 micrograms/ml were used; per os administration of etoposide at 25 mg x 2/day has been shown to produce a continuous plasma concentration of etoposide of around 1 microgram/ml. The results were as follows: 1) The IC50 of CDDP after 72 hours of exposure was 8.0 micrograms/ml and that of etoposide after 114 hours was 3.0 micrograms/ml. 2) After 100 hours of exposure to 1 microgram/ml of etoposide, cell cyclic phase analysis showed cells predominantly in G2/M phase arrest. 3) After 24-hour administration of CDDP, it was more than 24 hours before a cytocidal effect was observed. 4) During continuous exposure of SHIN-3 to etoposide (1 microgram/ml) for 6 days, CDDP was added on the 1st, 3rd, and 5th days. The largest ratio of growth inhibition with combined treatment to that with CDDP alone was attained on the 5th day. We conclude that, in combination regimens using low-dose continuous etoposide, CDDP should be added after etoposide administration is begun. PMID- 8429247 TI - [Glycosphingolipids of human gynecological malignant cell lines in vitro]. AB - We focused on the glycosphingolipids as one of the various biological phenotypes on the cell membrane of malignant tumor cells. The composition of three glycosphingolipids in seven human cell lines, derived from gynecological malignant tumors, was analyzed biochemically and immunochemically. Among the seven human cell lines, lactosylceramide sulfate was characteristically detected in these endometrial cancer cell lines, HEC-108, SNG-II, and SNG-M. In contrast, no sulfoglycolipids were found in the other malignant cell lines. HEC-108, which forms a moderately differentiated endometrial adenocarcinoma in nude mice, had a much higher ratio of lactosylceramide sulfate/GM3 than SNG-II and SNG-M cell lines, both of which grow as poorly differentiated endometrial adenocarcinoma in nude mice. TLC-immunostaining revealed that the sialyl type I carbohydrate chain with polysialyl groups was preferentially contained in the two sarcoma-derived cell lines and not in the five carcinoma-derived ones, while the type I carbohydrate chain was detected in all lines examined. Furthermore, the carcinoma cell lines were found to express A,B,H,Le(a),Le(b),Le(x) and Le(y) glycolipids, whereas the sarcoma cell lines scarcely expressed them. These results suggest that the expression of lactosylceramide sulfate is one of the biological characteristics of the three endometrial cancer cell lines, and the fucosylation to terminal residues of the type I and type II carbohydrate chains might be reduced in uterine sarcoma- and ovarian sarcoma-derived cell lines. PMID- 8429248 TI - [Evaluation of routine gas determinations in umbilical cord blood at cesarean section]. AB - From 1985 to 1989, analysis of the umbilical cord blood gas in 546 cases of cesarean section delivery under spinal anesthesia was carried out to evaluate its relationship with neonatal asphyxia, and the following results were obtained. (1) The UApH of a normal newborn with an Apgar score > 7 was 7.28 +/- 0.04. Acidosis occurred in 33.3% of the infants born with an Apgar score < 7, but the incidence was only 3.9% in those with an Apgar score > 7. Sixteen% of the acidosis was metabolic and 84% belonged to the mixed type. The UApH and Apgar scores were most significantly related. (2) The abnormal UVpH was 4.9% (< M - 2SD). (3) There was no significant difference in UApH between cesarean section performed before and after the onset of labor. (4) The short term morbidity included: one death, 2 cases of convulsions, 2 cases of cerebral edema, and one case of periventricular leukomalacia. (5) The long term morbidity in the 29 cases referred to NICU included: 18 cases of normal growth, 2 cases of severe cerebral palsy, 3 cases of mild cerebral palsy, and 6 cases of mental retardation. (6) In view of the long term morbidity of the newborn, as seen in the correlation between the Apgar score and UApH, it was considered that, the critical points in neonatal asphyxia were Apgar score < or = 3 and UApH < 7.10. My results suggested that the umbilical cord blood gas analysis was helpful for the diagnosis of neonatal asphyxia, and the prognosis of the newborn. PMID- 8429249 TI - [A case of huge ovarian tumor]. PMID- 8429250 TI - [Two cases of atypical polypoid adenomyoma of the uterus]. PMID- 8429251 TI - [Pregnancy and delivery of a 41-year-old chronic hemodialysis patient: a case report]. PMID- 8429252 TI - [Fetal heart rate monitoring during maternal swimming]. AB - Fetal physiology during swimming in pregnancy has remained unclear. To assess the fetal status, the fetal heart rate (FHR) was measured during maternal swimming. We studied individually a group of seventeen women in normal pregnancy (during their third trimester, 35-38th week) who participated in a maternal swimming class. We adapted the Doppler ultra-sound transducer for underwater use and attached it to the mothers' abdomen. FHR was recorded before, during and after swimming. The group swam from 375 to 750 meters in 33 to 41 minutes. During swimming, motion artifacts interfered with the FHR signal and we were only able to detect FHR in eleven in the group. The mean FHR rose significantly during swimming in eight of eleven women compared to the FHR recorded before the exercise. The FHR pattern was reactive and baseline variability was preserved in all cases. No pathological deceleration was seen. It is concluded that maternal swimming under such class conditions is safe for the fetus. PMID- 8429253 TI - [Pharmacokinetics of CDDP by the route of administration in patients with ovarian cancer]. AB - CDDP was administered to 36 patients with ovarian cancer by one of three routes: i.v., ip, or ia, and its pharmacokinetics was studied. 1) With an initial dose of 100 mg/body the peak values for blood free-pt were 1.63 +/- 0.78 micrograms/ml (S.D.) in the i.v. group, 0.61 +/- 0.53 micrograms/ml (S.D.) in the ip group, and 1.62 +/- 0.78 micrograms/ml (S.D.) in the ia group. The area under the curve (AUC) was 2.56 +/- 0.72 micrograms.hr/ml (S.D.), 1.84 +/- 0.75 micrograms.hr/ml(S.D.), and 2.70 +/- 0.51 micrograms.hr/ml (S.D.), respectively. 2) The free-pt level of the ascitic fluid in the ip group was 25.3 +/- 28.1 micrograms/ml (S.D.) just after the injection of 100 mg/body. 3) The ovarian tissue level following the initial CDDP dose of 100 mg/body was the highest in the ia group, next highest in the ip group, and lowest in the i.v. group. In some cases, the ovarian tissue level was markedly elevated after repeated ip injections. Deep-middle layer concentrations in ovarian tissue were not so low in the ip group as in the i.v. or ia group. As noted above, the blood free-pt peak was low following ip administration, but the ovarian tissue level was higher following ip administration than following i.v. though lower than following ia administration. These findings suggest that ip administration of CDDP may be useful in treating large tumor masses. Good results are expected particularly following repeated ip therapy, which sometimes provided a markedly high tissue level. PMID- 8429254 TI - Effect of phosphatidylcholine molecular species on the uptake of HDL triglycerides and cholesteryl esters by the liver. AB - It has previously been shown that the hydrolysis of high density lipoprotein (HDL) phospholipids by hepatic lipase promotes the hepatic uptake of triglyceride and cholesteryl ester from HDL. Since the hydrolysis of HDL phospholipids promotes HDL cholesteryl ester uptake, then it is possible that the hepatic metabolism of HDL could be altered by changing the molecular species composition of HDL phosphatidylcholine (PC) (the major phospholipid in HDL). To test this possibility the uptake of [3H]triolein and [14C]cholesteryl oleate by the isolated perfused rat liver was determined in a nonrecirculating perfusion system following a bolus injection of reconstituted HDL (rHDL) prepared with rat serum HDL apolipoproteins, [3H]triolein, [14C]cholesteryl oleate, unesterified cholesterol, and one of five molecular species of phosphatidylcholine (16:0-18:2, 16:1-16:1, 18:0-18:2, 18:1-16:0, or 20:1-20:1), 18:1-16:0 phosphatidylcholine diether, or the total phosphatidylcholine fraction isolated from rat serum HDL. The apolipoprotein profiles and lipid compositions of all the rHDL were similar. The greatest uptake of [3H]triolein was obtained with the rHDL prepared with 16:0 18:2 phosphatidylcholine. The amount of [3H]triolein taken up by the liver when the rHDL were prepared with 16:1-16:1, 18:0-18:2, 18:1-16:0, 20:1-20:1, 18:1-16:0 phosphatidylcholine diether, or rat serum HDL phosphatidylcholine was 56.7%, 51.7, 39.9%, 27.6%, 31.8%, and 84.7%, respectively, of the amount taken up from the rHDL prepared with 16:0-18:2 phosphatidylcholine. Likewise, the greatest amount of [14C]cholesteryl oleate was taken up by the liver when the rHDL was prepared with 16:0-18:2 phosphatidylcholine and the variation in [14C]cholesteryl oleate uptake from the various rHDL displayed a pattern of dependence on the phosphatidylcholine molecular species of rHDL similar to triolein uptake. The variation in the rate of rHDL phosphatidylcholine hydrolysis by hepatic lipase in vitro as a function of the PC molecular species composition of the rHDL also resembled the variation in the hepatic uptake of [14C]cholesteryl oleate and [3H]triolein. Partial substitution of rHDL phosphatidylcholine with the minor phospholipids isolated from rat serum HDL affected neither the hepatic uptake of [3H]triolein or [14C]cholesteryl oleate nor phosphatidylcholine hydrolysis by hepatic lipase in vitro when 16:0-18:2 phosphatidylcholine or rat serum HDL phosphatidylcholine was used to prepare the rHDL. On the other hand, when rHDL was prepared with 20:1-20:1 phosphatidylcholine, the inclusion of the minor phospholipids increased [3H]triolein and [14C]cholesteryl oleate uptake and phosphatidylcholine hydrolysis approximately twofold.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429255 TI - Impaired discrimination between stereoisomers of alpha-tocopherol in patients with familial isolated vitamin E deficiency. AB - We assessed whether patients with familial isolated vitamin E deficiency could discriminate between natural (RRR-) and synthetic (SRR-) stereoisomers of alpha tocopherol labeled with six (d6) or three (d3) deuterium atoms, respectively. After oral administration of 20 mg of each of the stereoisomers, patients (seven) and controls (seven) had similar concentrations of both in chylomicrons, similar initial increases of both, and similar rates of decrease of d3-SRR-alpha tocopherol in plasma. Patients and controls differed in their abilities to maintain plasma d6-RRR-alpha-tocopherol concentrations. Controls maintained plasma d6-RRR-alpha-tocopherol concentrations by preferentially secreting it in very low density lipoprotein (VLDL). Three of seven patients did not discriminate between the two stereoisomers and their plasma and lipoprotein d6-RRR-alpha tocopherol concentrations declined rapidly. The remaining patients were intermediate between non-discriminators and controls in their ability to discriminate and maintain plasma d6-RRR-alpha-tocopherol concentrations. The degree of discrimination between the two stereoisomers in the patients was correlated with the age of onset of the neurologic disability (r2 = 0.64, P < 0.03). Estimates based on the rate of decrease of plasma d6-RRR-alpha-tocopherol in non-discriminators suggest that the entire plasma alpha-tocopherol pool of normal subjects is replaced daily. We suggest 1) that a hepatic alpha-tocopherol binding protein, which preferentially incorporates RRR-alpha-tocopherol into VLDL, is required to maintain plasma RRR-alpha-tocopherol concentrations; 2) that non-discriminators are lacking this protein, or have a marked defect in the RRR alpha-tocopherol binding region of the protein; and 3) that patients who discriminate, but have difficulty maintaining plasma RRR-alpha-tocopherol concentrations, have a less severe defect, or perhaps a defect in the transfer function of the protein. PMID- 8429256 TI - Lipid vesicle fusion induced by phospholipase C activity in model bile. AB - Using a system of phosphatidylcholine-cholesterol vesicles to model the vesicle phase of mammalian bile (1:1 molar ratio) we evaluated whether very small amounts of C. perfringens phospholipase C activity (0.5-6.5 nmol/min per ml) could lead to vesicle fusion, a precursor step for cholesterol precipitation in gallbladder bile. Quasielastic light scattering spectroscopy (QLS) was used to monitor vesicle growth and aggregation in model bile (0.89 mM total lipid) in the presence of phospholipase C. Vesicle growth over 2 h could be detected with phospholipase activity as little as 0.5 nmol/min per ml. Vesicle growth was sustainable over days in the absence of Ca2+ once as little as 3-7 mol% diacylglycerol had been generated as a result of the initial phospholipase C treatment. The presence of fusion intermediates was confirmed using transmission electron microscopy. In addition, kinetically slow vesicle fusion with intravesicle content mixing and minimal leakage was also confirmed by fluorescence spectroscopy using two populations of vesicles containing 5 mM TbCl3 or 50 mM dipicolinic acid. Efficient fusion (40% maximum fluorescence) was obtained at 30 min at 25 degrees C with phospholipase C activity. This level of enzyme activity approximates that found in human gallbladder bile (1.2 nmol/min per ml). We conclude that the hydrolysis products of phospholipase C activity can, in very small amounts (3-7 mol% diacylglycerol), lead to destabilization and fusion of cholesterol-saturated biliary vesicles. A reappraisal of the importance of phospholipase C hydrolysis products in the pathogenesis of cholesterol gallstones is warranted based on these observations. PMID- 8429257 TI - Regional differences in triacylglycerol synthesis in adipose tissue and in cultured preadipocytes. AB - The initial suspicion that obesity increases coronary risk has been much sharpened with the demonstration that risk is more tightly linked to abdominal than to peripheral obesity, and tighter yet again when the mass of omental adipose tissue is taken into account. These data suggest that important metabolic differences might exist between adipocytes from different regions, and indeed, it has long been appreciated that triacylglycerol hydrolysis can be stimulated to a greater extent in omental than in subcutaneous adipocytes. The present study focuses on triacylglycerol synthesis in human subcutaneous and omental adipocytes, a process which, by contrast, has received relatively little attention. Experiments were done on adipose tissue removed at laparotomy and on cultured preadipocytes. With the former, triacylglycerol synthesis was measured in the presence and absence of oleate added to the medium using radiolabeled glucose and oleate as tracers. The results demonstrate that under all conditions examined triacylglycerol synthesis in subcutaneous adipose tissue exceeded that in deep omental adipose tissue. To study the cells in more detail, preadipocytes were cultured and triacylglycerol synthesis was examined again under basal conditions and with stimulation with insulin and acylation stimulating protein (ASP). Under basal conditions, particularly when oleate was added to the medium, clear differences were present such that triacylglycerol synthesis was substantially greater in subcutaneous preadipocytes than in omentally derived preadipocytes. These differences were more pronounced when the cells were stimulated with either insulin or acylation stimulating protein. Overall, triacylglycerol synthetic capacity in subcutaneous tissue exceeded that in omental tissue. As a consequence, omental tissue as compared to subcutaneous adipose tissue would have a limited capacity to prevent fatty acids from reaching the liver and stimulating hepatic lipoprotein synthesis. PMID- 8429258 TI - Spinal cord of the rat contains more lipoprotein lipase than other brain regions. AB - Lipoprotein lipase (LPL) is important for the delivery of triglyceride fatty acids (TGFA) to a variety of tissues. We have used measurements of heparin releasable LPL activity, immunohistochemistry, in situ hybridization, and Northern analysis to more fully characterize the location of LPL within the entire central nervous system (CNS) of the rat. Surprisingly, the levels of LPL activity and mRNA in the caudal spinal cord were 5- to 10-times higher than those found in any other area of the brain, levels similar to those found in adipose tissue or skeletal muscle. A number of cell types including neurons in the hippocampus, Purkinje cells of the cerebellum, and cells deep within the cortex were identified as sources of LPL mRNA. LPL protein was found within many vascular structures throughout the CNS, and within Purkinje cells. The strongest immunostaining was around nerve rootlets associated with the caudal spinal cord. Feeding studies were carried out with [14C]oleic acid to see whether CNS LPL functioned in the uptake of TGFA. These studies demonstrated uptake of chylomicron triglyceride fatty acids throughout the CNS. The localization of LPL within these structures suggests that the uptake of triglyceride fatty acids is an integral part of normal lipid metabolism of the central nervous system and may be important in regulating feeding behavior and/or maintaining normal neuronal function. PMID- 8429259 TI - Role of thyroid hormone in the expression of apolipoprotein A-IV and C-III genes in rat liver. AB - The genes coding for apolipoproteins A-I, C-III, and A-IV are closely linked to one another in the rat genome. Thyroid hormone stimulates apoA-I expression in rat liver by an unusual mechanism that enhances the maturation of mRNA. This hormone also increases apoA-IV mRNA abundance by a mechanism not yet studied, and its role in the expression of apoC-III has not been defined but may be of relevance to the metabolism of triglyceride-rich lipoproteins. We therefore measured the transcriptional activity of the apoA-IV and apoC-III genes and the abundance of their nuclear RNA and total cellular mRNA in livers of control rats and rats made hyper- and hypothyroid. After a single receptor-saturating dose of triiodothyronine (3 mg/100 g body weight), apoA-IV gene transcription increased at 20 min and reached a maximum of 260% of control at 6 h. Increases of transcription were reflected in increases of nuclear and total apoA-IV mRNA levels. ApoC-III gene transcription was temporarily increased to 160% at 2 h without changes in the abundance of its nuclear or total mRNA over 24 h. Lower hormone doses (20-500 micrograms/100 g body weight) stimulated apoA-IV mRNA transcription as well, but tended to reduce transcription from the apoC-III gene. Upon chronic administration of thyroid hormone, apoA-IV transcription decreased to 55% and nuclear apoA-IV RNA levels to 87% of control. However, total cellular apoA-IV mRNA levels increased to 279% of control, implying stabilization of mRNA in the cytoplasm. ApoC-III transcription decreased to 28% of control, but abundance of nuclear and total cellular apoC-III mRNA was reduced to a lesser extent. In hypothyroid rats, apoA-IV gene expression was decreased fourfold at the transcriptional level. In contrast, apoC-III gene transcription increased to 178% of control, but the abundance of nuclear and total cellular apoC-III mRNA did not differ from control rats. Thus, thyroid hormone affects the abundance of apoA-IV mRNA by changing its synthesis and its rate of degradation and enhances the efficiency of apoC-III mRNA maturation, thereby blunting the net effect of altered mRNA synthesis on mRNA abundance. PMID- 8429260 TI - Apolipoprotein E enhances lipid exchange between lipoproteins mediated by cholesteryl ester transfer protein. AB - The direct effect of apolipoprotein (apo) E on cholesteryl ester transfer protein (CETP) activity was studied using lipoproteins from a subject with apoE deficiency (Atherosclerosis. 1991. 88: 15-20) as a model system. The transfer of cholesteryl ester (CE) from discoidal bilayer particles (DBP) to very low density lipoprotein (VLDL) was enhanced by incorporation of apoE into VLDL. This enhancement was induced only in the presence of CETP activity. Moreover, after incubation of CETP with VLDL, CETP activity and immunoreactivity were coeluted with apoE-incorporated VLDL (E-VLDL) on a gel filtration column (Sephadex G-150), but there was little CETP activity and immunoreactivity with apoE-free VLDL (C VLDL), suggesting that E-VLDL had higher affinity for CETP compared to C-VLDL. The supplementation of the apoE-deficient serum with apoE enhanced the CETP mediated changes of amount of CE and triglyceride (TG) in the high density lipoprotein (HDL) fraction, which were completely inhibited by the addition of the monoclonal antibody against CETP that blocks CETP activity. Our results suggest that 1) apoE enhances the cholesteryl ester and triglyceride transfer between VLDL and HDL via cholesteryl ester transfer protein, and 2) this effect of apoE may be mediated by enhancing the affinity of CETP for VLDL. PMID- 8429261 TI - Prevalence of familial hypercholesterolemia among young north Karelian patients with coronary heart disease: a study based on diagnosis by polymerase chain reaction. AB - Two deletions of the low density lipoprotein (LDL) receptor gene account for about 90% of the mutations that cause familial hypercholesterolemia (FH) in eastern Finland. The FH-Helsinki mutation deletes exons 16, 17 and a portion of exon 18, while the FH-North Karelia allele is characterized by a deletion of seven nucleotides from exon 6 of the LDL receptor gene. We developed a DNA assay based on the use of polymerase chain reaction (PCR) which simultaneously detects both of these mutations. We have screened 90 young (< 45 years) eastern Finns with symptomatic coronary heart disease (CHD) for the presence of these FH genes. One or the other of the mutations was present in 4 out of 55 survivors of acute myocardial infarction (AMI) and 4 out of 35 patients with angina pectoris (AP), but in none of 50 healthy controls of similar age. These data show a relatively high prevalence of confirmed FH in young CHD patients (AMI and MI combined: 8/90, or 9%), and also demonstrate the feasibility of PCR techniques in diagnosis of FH among populations with enrichment of specific types of LDL receptor gene mutations. PMID- 8429262 TI - In vivo evidence that the lipid-regulating activity of the ACAT inhibitor CI-976 in rats is due to inhibition of both intestinal and liver ACAT. AB - CI-976, a new trimethoxy fatty acid anilide, is a potent and specific inhibitor of liver and intestinal acyl coenzyme A:cholesterol acyltransferase (ACAT) in vitro. Several in vivo approaches were used to determine the efficacy and sites of action of this compound in rats. CI-976 decreased non-high density lipoprotein (HDL)-cholesterol and increased HDL-cholesterol in rats with pre-established dyslipidemia. High performance gel chromatographic separation of plasma lipoproteins also revealed that CI-976, but not CL 277,082, lowered low density lipoprotein (LDL)-cholesterol and elevated HDL-cholesterol. Bay o 2752, octimibate, melinamide, and SaH 58-035 were all less potent in vivo compared to CI-976 and CL 277,082, and CI-976 produced the greatest decrease in liver cholesteryl esters. Subcutaneous (SC) administration of CI-976 was also efficacious in cholesterol-fed animals. In sucrose-fed rats, oral and SC CI-976 administration potently lowered plasma triglycerides. Hepatic cholesteryl ester accumulation in the ethinyl estradiol-treated rat was also diminished by orally administered CI-976. ACAT activity and cholesteryl ester mass were dose dependently decreased in the livers from cholesterol-fed rats treated with CI 976, suggesting a direct effect on the liver. In both hypercholesterolemic and hypertriglyceridemic models, CI-976 also decreased plasma apoB concentrations. In other experiments radiolabeled CI-976 accumulated in the liver after multiple doses. Time-dependent changes in biliary lipid and bile acid secretion suggested that free cholesterol did not accumulate in the liver but instead was excreted as such or as bile acid. Finally, inhibition of endogenous and exogenous intestinal cholesterol absorption was demonstrated using several in vivo techniques. The combined data strongly supports the hypothesis that orally administered CI-976 inhibits both intestinal and hepatic ACAT, and that both of these enzymes may be determinants of plasma lipid concentrations in the rat. PMID- 8429263 TI - Dense low density lipoprotein subspecies with diminished oxidative resistance predominate in combined hyperlipidemia. AB - Patients presenting combined hyperlipidemia (CHL) display an elevated risk of coronary heart disease. The atherogenic lipoprotein particles implicated in this disorder remain ill defined. We determined the qualitative and quantitative characteristics of the density distribution of low density lipoprotein (LDL) particle subspecies in nine subjects defined phenotypically as presenting CHL, and under strict dietary control. Seven CHL patients possessed familial antecedents of premature coronary heart disease; none were E2E2 homozygotes. Five LDL subspecies were isolated by density gradient ultracentrifugation in the density range 1.019-1.063 g/ml. In all patients, the LDL profile was skewed towards the dense subspecies (LDL-4, d 1.039-1.050 g/ml and LDL-5, d 1.050-1.063 g/ml), representing 47% of total LDL mass; by contrast, these subspecies accounted for only 30% of LDL mass in five normolipidemic subjects (P < 0.01). In addition, plasma LDL mass concentrations were some twofold higher in CHL patients as compared to normolipidemic subjects. The % mass of LDL-4 was positively correlated with plasma triglyceride and apoB levels. LDL-2 and LDL-3 in CHL patients were triglyceride-enriched (11.9 and 7.2%, respectively) as compared to the corresponding subspecies in normolipidemic subjects (6.6 and 3.7%, respectively; P < 0.05 in each case). LDL particle size decreased with increase in density in both groups; however, significant differences were found between corresponding LDL subspecies (LDL-1, -3, -4, and -5) in CHL patients and normolipidemic subjects, a finding suggestive of dissimilar molecular organization, despite correspondence in hydrated density. The copper-induced oxidative modification of LDL subspecies was assessed by determination of conjugated diene formation. In both groups, LDL-5 was distinct in exhibiting a marked diminution in oxidative resistance as indicated by a significant reduction (P < 0.01) in mean lag time. The oxidative susceptibility of LDL subspecies in both groups was independent of vitamin E content when expressed as the ratio vitamin E/LDL mass, although dense LDL in CHL patients tended to be deficient in this antioxidant. The diminished oxidative resistance of dense LDL subspecies could not be accounted for by enrichment in polyunsaturated fatty acids in either group. These studies suggest that in consequence of their elevated circulating concentration and diminished oxidative resistance, dense LDL subspecies represent putative atherogenic subspecies in combined hyperlipidemia. PMID- 8429264 TI - Apolipoprotein A-I-cell membrane interaction: extracellular assembly of heterogeneous nascent HDL particles. AB - The ability of lipid-free human apoA-I expressed by transfected Chinese hamster ovary (CHO) cells to form apoA-I-lipid complexes extracellularly when incubated with CHO cell monolayers was investigated. Lipid-free apoA-I was incubated with nontransfected CHO-C19 cells for 24 h and extracellular assembly products were isolated at d < or = 1.235 g/ml; approximately 12% of the incubated apoA-I floated at d < or = 1.235 g/ml when apoA-I was added at 10 micrograms/ml. The composition of the particles was 51.3% protein, 20.3% phospholipid, and 28.3% cholesterol. Electron microscopy of the apoA-I-lipid complexes revealed that discoidal particles 15.4 +/- 4.1 nm diameter predominated but some vesicular particles 34.7 +/- 16.8 nm diameter were also in evidence. Nondenaturing gradient gel electrophoresis of the extracellular assembly products formed after 24 h incubation with 10 micrograms/ml apoA-I showed particle size heterogeneity with major bands at 11.2 and 9.0 nm; additional minor components banded at 7.3, 17.7, and 19.5 nm. This size distribution, as well as composition and electron microscopic structure, is similar to that of complexes isolated from the medium of CHO cells transfected with the human apoA-I gene. The formation of extracellular assembly complexes was concentration-dependent such that at 2 micrograms apoA-I/ml for 24 h, primarily 7.3 nm complexes were formed; at 4 micrograms/ml the distribution was more heterogeneous and the major band peaked at 9.2 nm, while at 8 micrograms/ml the 7.3 nm component was greatly diminished and the 11.2 nm component was the major one.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429265 TI - New model to study cholesterol uptake in the human intestine in vitro. AB - A new model to study cholesterol uptake in the human intestine in vitro is described. Human small intestine organ cultures were incubated with mixed micelles containing bile acid, phospholipid, and cholesterol or its nonabsorbable analogue, sitosterol; trace amounts of labeled cholesterol or sitosterol were added to the micelles. After incubation, the lipids were extracted from the cells and cholesterol and sitosterol uptake was evaluated. Specific cholesterol uptake was determined as a difference between cholesterol and sitosterol uptake. Cholesterol, but not sitosterol, uptake was time- and dose-dependent. Rapid and slow phases of cholesterol uptake were observed. Cholesterol uptake was also temperature-dependent. Removal of epithelial cells from human intestine explants reduced cholesterol, but not sitosterol, uptake. Inhibition of acyl CoA:cholesterol acyltransferase by Sandoz compound 58-035 and treatment with monensin reduced cholesterol uptake, but not sitosterol uptake, in a dose dependent manner. In contrast, treatment of cultures with an inhibitor of 3 hydroxy-3-methyl-glutaryl coenzyme A reductase, lovastatin, stimulated cholesterol, but not sitosterol, uptake in a dose-dependent manner; mevalonic acid reversed the effect of lovastatin. The presented model allows large-scale in vitro studies of different stages of cholesterol absorption in the human intestine. PMID- 8429266 TI - Oestradiol-17 beta modulates the actions of pharmacologically distinct forms of protein kinase C in rat anterior pituitary cells. AB - Phorbol ester-induced release of LH and GH from rat anterior pituitary tissue in vitro is differentially inhibited by some, but not other, inhibitors of protein kinase C (PKC), suggesting that pharmacologically distinct species of PKC may have different functional roles in these cells. Since stimulus-induced anterior pituitary hormone release can be enhanced by oestradiol-17 beta (OE2) pretreatment, we investigated the effect of OE2 treatment of long-term (4 weeks) ovariectomized rats on the amount, activity and cellular actions of pharmacologically distinct PKC species in rat anterior pituitary tissue. Here we report that OE2 treatment enhanced phorbol 12,13-dibutyrate (PDBu)-induced LH but not GH release measured in vitro. This effect of OE2 on LH release may involve synthesis of additional PKCs that are not targeted by the synthetic diacylglycerol, 1,2-dioctanoyl-sn-glycerol (DOG). Measurements of anterior pituitary PKC activity and [3H]phorbol ester-binding studies suggested that the facilitatory action of OE2 on LH release may occur, at least in part, by altering the quantity and activity of PKC(s). Our results also demonstrate that the OE2 induced PKC(s) which facilitate LH release may be of the type that are not dependent upon raised intracellular Ca2+ for their activation and display distinct pharmacological properties (being readily activated by PDBu, but not by DOG, and are staurosporine-sensitive but H7-insensitive). This facilitatory action of OE2 on PKC-induced LH release does not appear to involve OE2-induced changes in the affinity of existing PKC(s) for PDBu, or changes in the amount of releasable LH in the pituitary prior to the stimulus. PMID- 8429267 TI - Bradykinin and vasopressin activate phospholipase D in rat Leydig cells by a protein kinase C-dependent mechanism. AB - In the present study we report that bradykinin stimulated phospholipase D activity in rat Leydig cells. Bradykinin added for 8 min stimulated choline formation in a dose-dependent manner and, in the presence of ethanol, bradykinin (100 nmol/l) stimulated transphosphatidylation by phospholipase D resulting in the formation of phosphatidylethanol. This stimulation was abolished after down regulation of protein kinase C by long-term pretreatment for 22 h with phorbol 12 myristate 13-acetate (PMA). The stimulation of phospholipase D by the simultaneous addition for 8 min of maximum concentrations of PMA and vasopressin (AVP), PMA and bradykinin, or AVP and bradykinin produced no additive phosphatidylethanol or choline response, suggesting that AVP, bradykinin and PMA stimulated phospholipase D-catalysed phosphatidylcholine hydrolysis by a similar protein kinase C-dependent mechanism. Furthermore, LH (10 ng/ml), insulin (500 nmol/l), GH (100 ng/ml), interleukin-1 beta (5 U/ml) and platelet-activating factor (200 nmol/l) were found not to activate phospholipase D, whereas the Ca2+ ionophore A23187 (10 mumol/l) stimulated phosphatidylethanol formation, suggesting that Ca2+ might be a regulator of phospholipase D in Leydig cells. PMID- 8429268 TI - Oestradiol negative feedback inhibition on LH secretion during lactation is prolonged in adolescent primiparous rhesus monkeys. AB - Lactational infertility in rhesus monkeys is significantly prolonged in adolescent primiparous compared with adult multiparous mothers. In order to determine if this longer period of infertility for young mothers is the result of a greater sensitivity to nursing-induced inhibition of LH release either by enhanced oestradiol negative feedback or a direct non-gonadally mediated suppression, the effects of periodic administration of oestradiol on serum LH concentrations in nursing ovariectomized adolescent primiparous (Prp; n = 5) and adult multiparous (Mlt; n = 7) mothers was assessed. Females were treated every 5 6 weeks with a 21-day time-release capsule of oestradiol which produced serum concentrations of approximately 250, 90 and 45 pmol/l by +6, +13 and +20 days after treatment. Thus, the design permitted assessment of LH and prolactin concentrations under a regime of 21 days of decreasing oestradiol levels followed by 2-3 weeks of no oestradiol treatment. Females were studied from week 3 to week 42 post partum and oestradiol treatment occurred during weeks 5, 11, 21, 26, 31, 36 and 41. Behavioural observations indicated that the amount of time mothers nursed their offspring decreased in a similar fashion throughout the lactational period for both Prp and Mlt females. LH concentrations under the 'no oestradiol' conditions progressively increased throughout lactation reaching maximum levels by week 36 post partum in a similar manner for both Prp and Mlt mothers. These data suggest that differences in fertility between adolescent and adult nursing mothers observed previously cannot be attributed to a difference in a direct non gonadally-mediated affect on LH. With respect to oestradiol negative feedback inhibition of LH, oestradiol treatment effectively suppressed serum LH concentrations at all points during lactation up to week 31, at which time LH concentrations were maximally suppressed in both Prp and Mlt mothers at +6 days after treatment but by day +13 LH values were significantly higher in Mlt females. After oestradiol treatment during week 36, LH values were again maximally suppressed on day +6 in Prp females but not in Mlt females. However, the response of serum LH after oestradiol treatment during week 41 was similar between Prp and Mlt females at all sampling points.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429269 TI - The ability of steroid-free bovine follicular fluid to suppress FSH secretion and delay ovulation persists in heifers actively immunized against inhibin. AB - Previous reports indicate that administration of steroid-free bovine follicular fluid (bFF) to intact heifers suppresses plasma FSH levels and delays the timing of ovulation. In addition, cessation of bFF treatment is associated with a rebound hypersecretion of FSH. To test the hypothesis that these effects of bFF are mediated by inhibin, we have compared the responses to bFF treatment in heifers actively immunized against the N-terminal sequence of inhibin alpha subunit (bI alpha(1-29)Tyr30-ovalbumin) with those in ovalbumin-immunized controls. Oestrous cycles were synchronized, and inhibin-immune (n = 10) and control (n = 10) heifers were subdivided into two groups which received either 5 ml bFF (n = 5) or 5 ml bovine serum (n = 5) every 4 h for a 60 h period starting 8 h before prostaglandin (PG)-induced luteolysis. Blood was withdrawn every 4 h for 10 days starting 30 h before luteolysis and ovaries were examined daily by ultrasonography. Overall, mean ovulation rate in bI alpha(1-29)Tyr30-immunized heifers was 44% higher (P < 0.02) than in controls. Inhibin antibody titres tested in bI alpha(1-29)Tyr30-immunized heifers before (19 +/- 3%), during (19 +/ 3%) and after (20 +/- 3%) bFF treatment did not differ. In the pretreatment period (i.e. mid-luteal phase), plasma FSH levels were 32% (P < 0.05) higher in inhibin-immunized than in control heifers. During administration of bFF to control heifers, plasma FSH was suppressed to a level 40% lower than in serum treated heifers (P < 0.02). Unexpectedly, bFF suppressed FSH to a similar extent in inhibin-immunized heifers (37% lower than in the serum-treated group; P < 0.025). Similarly, a post-bFF rebound hypersecretion of FSH was observed in both control (P < 0.05) and inhibin-immunized (P < 0.05) heifers, although the FSH rebound lasted about 24 h longer in the latter group (P < 0.001). The timing of the preovulatory LH surge in control (86 +/- 7 h post-PG) and immunized (81 +/- 6 h post-PG) groups treated with serum was similar as was the timing of the preovulatory rise in plasma oestradiol and the subsequent rise in plasma progesterone. However, bFF treatment delayed (P < 0.001) the preovulatory surges of LH and oestradiol and the subsequent rise in plasma progesterone to a similar extent (> 4 days) in both control and inhibin-immunized groups.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429270 TI - Cell interactions in initiation of mammary epithelial proliferation by oestradiol and progesterone in prepubertal heifers. AB - The acute temporal effects of exogenous oestradiol (0.1 mg/kg per day), progesterone (0.25 mg/kg per day) or both together on the proliferative response of epithelial cells, fibroblasts, adipocytes and endothelial cells in the mammary tissue of prepubertal cross-bred heifers were determined. Mammary biopsies were taken immediately before, then 24, 48 and 96 h after the initiation of daily administration of hormones to three heifers per treatment group. Incorporation of [3H]thymidine into explants prepared from biopsies was evaluated after a 1-h incubation by measuring trichloroacetic acid (TCA)-insoluble radioactivity in explant homogenates as well as by quantitative histoautoradiography. Incorporation expressed as d.p.m./mg tissue or d.p.m./microgram DNA was increased (P < or = 0.05) approximately 11-fold by 96 h in oestradiol-treated heifers. Progesterone-treated animals were unresponsive and heifers treated with both hormones were intermediate in response compared with oestradiol-treated heifers. Autoradiographic data for ductal or terminal duct epithelial cells showed similar dramatic increases in labelling by 24 h with further increased (P < 0.01) labelling by 96 h (5.1% vs 0.1%) in heifers given oestradiol. As with incorporation, tissue from progesterone-treated heifers showed no time or treatment response compared with pretreatment biopsies and tissue from heifers given both showed intermediate responses, i.e. significantly increased labelling by 96 h compared with pretreatment (P < or = 0.05) but less labelling (P < 0.05) than oestradiol-treated heifers. A proliferative response of epithelial cells in oestradiol-treated heifers occurred prior to the response of fibroblasts adjacent to epithelial cells (< or = 50 microns).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429271 TI - Reproducibility of the cortisol response to stimulation with a low dose of ACTH(1 24): the effect of basal cortisol levels and comparison of low-dose with high dose secretory dynamics. AB - We compared the reproducibility and repeatability of the acute adrenal response to low doses (90 and 500 ng/1.73 m2) of Synacthen (ACTH(1-24)) with that of the standard dose (250 micrograms/1.73 m2). We also examined the effect of basal cortisol levels on peak values achieved after stimulation with a low dose. ACTH(1 24) was given to six male volunteers: 90 ng/1.73 m2 twice at 90-min intervals on day 1, and 90 and 500 ng/1.73 m2 once on day 2 and 250 micrograms/1.73 m2 once on day 3. The rise in serum cortisol concentration with repeated low doses of ACTH was not attenuated (161 +/- 49 (S.D.) nmol/l on initial vs 150 +/- 41 nmol/l on repeat stimulation; P = 0.5) and this was reproducible (161 +/- 49 nmol/l on day 1 vs 148 +/- 15 nmol/l on day 2; P = 0.6). A dose of 500 ng ACTH(1-24)/1.73 m2 produced a maximal adrenal response in that the rise in serum cortisol concentration at 20 min was identical with that produced at the same time by the standard dose of 250 micrograms/1.73 m2. There was a strong positive correlation between the basal cortisol level and peak cortisol concentration after low-dose ACTH stimulation (r = 0.93, P < 0.001) but not between the basal cortisol level and the incremental rise (r = -0.1, P = 0.69). These results suggest that the cortisol response to low-dose ACTH stimulation is reproducible and not attenuated by repeat stimulation at 90-min intervals.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429272 TI - Comparison of the effects of insulin-like growth factors-I and -II on the human osteosarcoma cell line OHS-4. AB - The effects of insulin-like growth factor-I (IGF-I) and IGF-II on the human osteoblast cell-line OHS-4 were investigated. Both IGF-I and IGF-II stimulated cell proliferation at nanomolar concentrations and alkaline phosphatase activity was decreased in a dose-dependent manner with either IGF-I or IGF-II. The production of the bone-specific protein osteocalcin was not influenced by either IGF-I or IGF-II. However, they acted synergistically with 1,25-dihydroxy-vitamin D3 at concentrations ranging from 10 to 100 nmol/l. Neither IGF-I nor IGF-II had an effect on either the basal or the parathyroid hormone-stimulated level of adenylate cyclase activity, and likewise they had no effect on phosphodiesterase activity. Binding and cross-linking experiments confirmed the presence of both type-I and type-II IGF receptors on the OHS-4 cells. The present study shows that IGF-I and IGF-II have similar effects on the parameters studied in these osteoblastic cells. They influenced both proliferation and differentiation markers. PMID- 8429273 TI - Russell's viper venom stimulates insulin secretion from rat islets of Langerhans. AB - Burmese Russell's viper venom (RVV) caused a dose- and temperature-dependent stimulation of insulin secretion from islets of Langerhans isolated from rat pancreas by collagenase digestion. RVV stimulated both basal and glucose-induced insulin secretion at concentrations which did not compromise islet cell viability as assessed by exclusion of trypan blue dye. The effects of RVV on insulin secretion could not be attributed to the activation of protein kinase C (PKC), since down-regulation of PKC by prolonged exposure to a tumour-promoting phorbol ester did not abolish subsequent secretory responses to RVV. However, RVV-induced insulin secretion was inhibited in the absence of extracellular Ca2+, and RVV did not stimulate insulin secretion from Ca(2+)-clamped electrically permeabilized islets at either substimulatory (50 nmol/l) or stimulatory (10 mumol/l) concentrations of Ca2+, suggesting that changes in cytosolic Ca2+ are important in the stimulation of insulin secretion by RVV. The phospholipase A2 (PLA2) inhibitor quinacrine caused a dose-dependent inhibition of RVV-induced insulin secretion, suggesting that the activation of PLA2, perhaps in response to Ca2+ influx, may be partially responsible for RVV-induced insulin secretion. PMID- 8429274 TI - Towards the pathogenesis of human pituitary tumours. PMID- 8429275 TI - Evidence that inhibin is an important factor in the regulation of FSH secretion during the mid-luteal phase in cows. AB - To investigate the physiological importance of inhibin in the regulation of FSH secretion in cows, seven cyclic cows were treated with an inhibin antiserum raised against bovine 32 kDa inhibin in a castrated goat. The same animals treated with a castrated goat serum (control serum) served as controls. On day 12 of the oestrous cycle (day 0 = day of oestrus), four of seven cows were injected with 100 ml inhibin antiserum first, and the remaining three cows with 100 ml control serum first. Twelve days after the second oestrus following the first serum injection (42-46 days after the first serum injection), the former four cows were injected with control serum and the latter three with inhibin antiserum. Follicular development after the injections of control serum or inhibin antiserum was assessed by daily ultrasonographic examination. Treatment with inhibin antiserum resulted in a marked increase (P < 0.01) in plasma concentrations of FSH and oestradiol-17 beta but not LH or progesterone, compared with those after treatment with control serum. Plasma concentrations of FSH increased significantly (P < 0.01) at 8 h after injection of anti-inhibin serum when compared with the control value. Concentrations of FSH in the plasma remained high for 72 h, then declined to the control level by 84 h, concomitant with an abrupt decrease in the titre of free inhibin antibody in the plasma. High concentrations of oestradiol-17 beta were observed between 36 and 96 h after treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429276 TI - Ontogeny and characterization of epidermal growth factor receptors on the fetal area of the sheep placenta. AB - Studies of the binding of 125I-labelled epidermal growth factor (EGF) to the ovine placenta were carried out on days 50, 90-100 and 140 of pregnancy. Membrane fractions were purified from the fetal area of the cotyledon. Two classes of binding sites were found. Their dissociation constants (Kd) were not significantly different for the three stages of pregnancy considered (high affinity Kd 54-70.2 pmol/l; low-affinity Kd 12.2 to 19 nmol/l). However, the number of high-affinity binding sites on days 90-100 was significantly (P < 0.01) greater (146 +/- 19 fmol/mg protein) than on either day 50 or day 140 (respectively 74.2 +/- 1.26 and 56.3 +/- 5.6 fmol/mg protein). Affinity cross linking studies followed by SDS-PAGE under reducing conditions demonstrated that the major part of the EGF was specifically cross-linked to a protein of molecular weight of 150 kDa and to lesser extent to 180 kDa and 130 kDa proteins. Membranes prepared from unfrozen tissues, in the presence of sodium iodoacetate to reduce endogenous enzymatic conversion of the 180 kDa form to the 150 and 130 kDa forms, still exhibited a major EGF-binding protein of 150 kDa. The occurrence of an increased number of EGF receptors at the period of rapid cotyledonary growth which coincides with the increase in placental hormonal secretions suggests that EGF has a role in the development of the ovine placenta. PMID- 8429277 TI - Circadian and daily rhythms of melatonin in the blood and pineal gland of free running and entrained Syrian hamsters. AB - The aim of this study was to develop a radioimmunoassay for the measurement of endogenous circulating melatonin concentrations in the Syrian hamster, and then to determine the effect of various photic manipulations upon this endocrine signal. In experiment 1, pineal-intact or pinealectomized adult male Syrian hamsters, housed under a long photoperiod (LD; 16 h light:8 h darkness) for 2 weeks, were then either maintained on LD or transferred to a short photoperiod (SD; 8 h light:16 h darkness) for a further 8 weeks. The profile of serum melatonin concentrations was determined from blood samples taken by cardiac puncture at intervals over a 24-h period. Radioimmunoassay revealed that daytime concentrations were at or below the limit of sensitivity of the assay (< or = 50 pmol/l). Under both photoperiods, the concentration of melatonin in the serum of pineal-intact animals rose 4-5 h after the onset of darkness, and the peak amplitude of the melatonin rhythm was not significantly different between the SD- or LD-housed animals (200-250 pmol/l). Premature exposure of animals to light during the dark phase of LD caused a precipitous decline in circulating concentrations to daytime values within 15 min and they remained there for several hours. In animals which experienced an uninterrupted night on either LD or SD, the most striking difference in the rhythm of endogenous melatonin secretion was the duration. Animals housed under LD had high levels until the start of the light period, a peak duration of 3.7 h.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429278 TI - Further studies on the mode of action of calcitonin on isolated rat osteoclasts: pharmacological evidence for a second site mediating intracellular Ca2+ mobilization and cell retraction. AB - Calcitonin is a circulating polypeptide that inhibits bone resorption by inducing both quiescence (Q effect) and retraction (R effect) in osteoclasts. Two structurally related members of the calcitonin gene peptide family, calcitonin gene-related peptide (CGRP) and amylin, inhibit osteoclastic bone resorption selectively via the Q effect. In the present study, we have made measurements of cell spread area in response to the application of amylin, CGRP and a peptide fragment of CGRP, CGRP-(Val8Phe37). We found that, over a wide concentration range (50 pmol/l to 2.5 mumol/l), the selective Q effect agonists did not produce an R effect. Furthermore, the peptides, when used at a 50-fold higher molar concentration than calcitonin, did not antagonize calcitonin-induced cell retraction. Additionally, experiments designed to measure changes in the intracellular free calcium concentration ([Ca2+]i) in single osteoclasts revealed that, unlike calcitonin, the non-calcitonin Q effect agonists did not produce a rise in [Ca2+]i. The peptides were also unable to attenuate the peak rise in [Ca2+]i induced by calcitonin. The results support our hypothesis that the inhibitory activity of calcitonin on osteoclastic bone resorption is mediated by two sites which may or may not be part of the same receptor complex. One of these is the classical Q effect site coupled to adenylate cyclase via a cholera toxin sensitive Gs. This site can be activated by nanomolar concentrations of calcitonin, amylin, CGRP or CGRP-(Val8Phe37). A novel R effect site, possibly coupled via a pertussis toxin-sensitive G protein to a [Ca2+]i elevating mechanism is predicted from this study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429279 TI - Induction of ornithine decarboxylase immunoreactivity in the male mouse kidney following testosterone treatment: an axial heterogeneity in the proximal tubule. AB - The effect of testosterone on the activity of ornithine decarboxylase (ODC), its protein level and immunocytochemical distribution were examined in the mouse kidney. Male BALB C mice at 8 weeks of age were used throughout. Fourteen hours before death, they received a subcutaneous injection of testosterone (1 mg/animal) or solvent to measure renal ODC activity or to detect the distribution of ODC immunoreactivity in the kidney. Renal ODC activity and the content of the enzyme were markedly increased after testosterone treatment. Histologically, few cells that were obviously immunoreactive to ODC were observed in the control animals and in the testosterone-treated animals a marked increase in ODC immunoreactivity was observed only in the cortex. ODC immunoreactive cells were located diffusely in the proximal tubule. In the pars recta, cells were stained weakly and homogeneously, while in the pars convoluta, the luminal surface of the cells showed stronger immunoreactivity. Moreover, many granule-like particles that were strongly ODC immunoreactive were observed inside the lumen of the pars convoluta. These results show that testosterone treatment induces an increase in ODC content in certain cells located in the proximal tubule of the cortex. PMID- 8429280 TI - Parathyroid hormone-related protein. PMID- 8429282 TI - Identification of alcoholic liver disease or hidden alcohol abuse in patients with elevated liver enzymes. AB - The aim of this prospective study was to identify alcoholic liver disease or covert alcohol abuse in unselected consecutive patients referred to a gastroenterologic out-patient's clinic for elevated liver enzymes. One-hundred and-thirteen patients were questioned about alcohol consumption, trauma history and loss of driver's licence. Laboratory tests claimed to reflect alcohol consumption and liver biopsy were taken. Using data from patients with the highest (26 patients) and lowest (29 patients) stated consumptions, logistic regression analysis identified violence score (trauma score + driver's licence score) and a laboratory index combining MCV, ASAT/ALAT ratio and IgA values as significant independent predictors of alcohol abuse (index = 2.3 x violence score +1.08 x laboratory index-3.19). Twenty-six patients openly admitted alcohol abuse (> 300 g week-1) and 85% of these had alcoholic liver damage. The index identified a further 12 patients as abusers who stated an intermediate alcohol consumption (25-300 g week-1). Half of these had alcoholic liver damage. Thus, a total of 34% of the patients were identified as abusers and in one-third of these the abuse was covert. PMID- 8429281 TI - Sex differences in essential hypertension. AB - A group of 41-year-old hypertensive men (n = 35, blood pressure (BP) 149.9 +/- 2.1/98.9 +/- 1.1 mmHg, mean +/- SEM) who had never received treatment for their condition were compared with hypertensive women of the same age (n = 18, BP 155.9 +/- 4.3/98.1 +/- 1.6 mmHg) with comparable body mass index (BMI, 25.9 +/- 0.5 vs. 24.9 +/- 4.5 kg m-2) who, also, had never received treatment. The lipid profile was more atherogenic in the men, with lower HDL cholesterol (1.21 +/- 0.04 vs. 1.38 +/- 0.06 mmol l-1, P = 0.04), higher total cholesterol (6.04 +/- 0.14 vs. 5.54 +/- 0.18 mmol l-1, P = 0.04) and triglycerides (1.80 +/- 0.16 vs. 0.96 +/- 0.10 mmol l-1, P < 0.001). The hypertensive men had higher haemoglobin (P < 0.001) and haematocrit. Plasma catecholamines were inversely related to BMI in the women only (r = -0.52, P < 0.05 for both noradrenaline and adrenaline). Women with BMI above 25 kg m-2 had significantly lower arterial plasma adrenaline and noradrenaline than those with BMI below 25 kg m-2 (28 +/- 5 vs. 78 +/- 16 pg ml 1, P < 0.01 and 101 +/- 17 vs. 206 +/- 33 pg ml-1, P < 0.01 respectively). A negative curvilinear relationship appeared between arterial adrenaline and insulin (r = 0.49, P = 0.05). These results suggest a male propensity for athero thrombogenic risk factors in otherwise comparable hypertensive subjects. A close relationship between metabolic risk factors within the normal range seems to exist even in hypertensive women. The decreased sympathetic activity at rest in the obese hypertensive women indicates different pathophysiological mechanism for hypertension in lean and obese. Decreased sympathetic activity and thus reduced energy expenditure, promotes a risk for weight gain, and could explain the inverse relationship between insulin and adrenaline. PMID- 8429283 TI - Exercise-provoked ST-segment depression and prognosis in patients recovering from acute myocardial infarction. Significance and pitfalls. AB - The importance of maximal versus submaximal exercise testing and the significance of heart failure on the prognostic value of exercise-provoked ST-segment depression > or = 0.1 mV was studied in 143 patients recovering from acute myocardial infarction. Patients were exercise tested prior to discharge and follow up lasted for up to 18 months (mean 17 months). End-point was first major event (i.e. first non-fatal reinfarction or death). A symptom-limited exercise test was superior to a heart-rate-limited test in detecting ST-segment depressions (27% vs. 20%: P < 0.5), and patients with ST-segment depression at lower heart rates did not have an increased risk of subsequent events compared with patients with ST-segment depression at higher heart rates (14% vs. 27%; NS). Heart failure surpassed ST-segment depression as a risk predictor (34% vs. 18%). Based on a meta-analysis including 13 studies (1987 patients) exercise-provoked ST-segment depression possessed an increased risk of subsequent major events (P < 0.0001; risk ratio = 1.90; 95% confidence limits 1.43,2.51). Thus, ST-segment depression provoked by a symptom-limited test selects patients with an increased risk of subsequent major events. In patients with a history of heart failure exercise-provoked ST-segment depression is of limited value. PMID- 8429284 TI - Does exercise-induced ST-segment depression predict benefit of medical intervention in patients recovering from acute myocardial infarction? The Danish Study Group on Verapamil in Myocardial Infarction. AB - The present study tested the hypothesis that the increased risk in patients with exercise-provoked ST-segment depression recovering from acute myocardial infarction could by abolished by anti-ischaemic medical intervention. Prior to discharge a symptom-limited exercise test was carried out. Patients were then double-blindly randomized to treatment with either verapamil 120 mg t.i.d. or placebo, and observed for up to 18 months (mean 17 months). End-point was first major event; i.e. non-fatal reinfarction or death. Two-hundred-and-ninety-eight patients were included. Forty-four patients with and 111 without exercise-induced ischaemia were randomized to verapamil and 39 and 104 respectively, to placebo. The overall number of events was 12.5%. In patients without ST-segment depression, 12.5% in the placebo group (hazard = 1) and 12.6% in the verapamil group (hazard = 1.13) had an event (NS). In patients with ST-segment depression 15.4% in the placebo group (hazard = 1.20) and 9.1% in the verapamil group (hazard = 0.85) had an event (NS). The latter reduction (41%) supports the hypothesis that patients with ST-segment depression, i.e. residual myocardial ischaemia, are those who benefit from anti-ischaemic intervention after myocardial infarction. PMID- 8429285 TI - Why should the relationship between diet and cancer be investigated in prospective studies? AB - Prospective studies on diet and cancer are needed for five reasons: (i) unlike xenobiotic agents which, if found to be carcinogenic, can at least in principle be dispensed with, food of some kind is indispensable; (ii) our present knowledge on the role of diet in cancer is limited and permits only tentative recommendations; (iii) plausible biological hypotheses for this role are available, as well as biological markers allowing their exploration in epidemiological studies; (iv) these studies demand a prospective approach--a number of prospective studies, each involving several tens of thousands of adults, are now being started in Europe: and (v) diet affects not only cancers but also a spectrum of other diseases, which need to be investigated in parallel, to acquire knowledge on which to base firm recommendations on a diet capable of maximizing benefits and minimizing the risk to health. PMID- 8429286 TI - The Malmo Diet and Cancer Study. Design and feasibility. AB - The Malmo Diet and Cancer study is a 10-year prospective case-control study in 45 64-year-old men and women (n = 53,000) living in a city with 230,000 inhabitants. One objective is to clarify whether a western diet is associated with certain forms of cancer whilst taking other life-style factors into account. Another broad question is whether oxidative stress and the activity in DNA-repairing systems influence the impact of diet on the development of all or certain forms of cancer. The study is also to act as a resource available for testing new hypotheses emanating from other studies. Initially food intake, heredity, socio economic factors, life-style pattern, occupational situation, previous and current diseases, symptoms and medications, will be determined. Viable lymphocytes, granulocytes, erythrocytes, and plasma/serum will be stored in a biological bank together with tumour specimens gathered from cases. The incidence and mortality of all cancer forms will then be followed for 10 years by existing registries. Data from the initial examination in these cases will then be compared with those of control subjects not having developed any form of cancer. A biomarker programme, utilizing the biological bank, has been developed and is aimed at finding predictors and/or precursors of cancer. A high participation rate (> 70%) and a high quality biological bank are prerequisites for a successful project. The experience gathered so far indicates that these goals are feasible. PMID- 8429287 TI - Dietary assessment methods evaluated in the Malmo food study. AB - A study to evaluate the relative validity of two dietary assessment methods, to be used in a prospective cohort study on diet and cancer, was conducted in a random sample of the Malmo population during 1984-1985. The study compared (i) a long, self-administered 'food-use questionnaire' (about 250 food items), asking for frequency of consumption of food and beverages and for usual portion size estimated with the help of a booklet of food pictures, and (ii) a combined method involving a shorter food frequency questionnaire (about 130 food items) and a 2 week diet diary, with (iii) a reference method requiring 18 days of weighed food records. The results indicated that subject compliance was quite good for both methods and, in particular, that the diet diary was well accepted. Both methods produced fairly good correlations with the reference method, of the order of 0.40 to 0.60 for most nutrients after adjustment for energy. Both methods tended to overestimate intake of fat, protein and carbohydrates as well as of vitamins and minerals. PMID- 8429288 TI - A biomarker for the assessment of niacin nutriture as a potential preventive factor in carcinogenesis. AB - The study of protective cellular responses to DNA damage has led to the working hypothesis that optimal niacin nutriture is a preventive factor in cancer. Described here is the development of a biomarker for determining niacin status termed Niacin Number. The combination of this biomarker with diet and cancer epidemiology will allow evaluation of the possible role of this nutrient in cancer risk. PMID- 8429289 TI - The Malmo biological bank. AB - Currently there are no available biological markers that have been satisfactorily validated clinically as 'intermediate end-points' for evaluation of the impact of diet on cancer. Therefore, there is high scientific value in banking biological specimens for future use in the Malmo Diet and Cancer Study. The banking procedures we have developed are adequate for assessing factors that estimate dietary intake (e.g. vitamins, lipids or micronutrients) as well as non-dietary variables (e.g. immunologic and ecogenetic factors of biologically effective doses of xenobiotic exposures) that may regulate the biological expression of dietary variables. This approach is further justified by state-of-the-art technology that includes assessment of the concept of gene-environment interactions. Purified viable mononuclear leucocytes, granulocytes, erythrocytes, plasma and serum are being stored from each individual entering the study under the guidelines of a strict quality control programme. Storage of tumour/normal tissues from incident cancer cases are also being stored and quality controlled. It is hoped that these banking procedures will encourage a high activity in biomarker development in the future. PMID- 8429290 TI - The Malmo biomarker programme. AB - Evaluation of pro-oxidant/antioxidant dietary factors in relation to individual susceptibility to cancer and cardiovascular disease is justified based on a review of the literature. This working hypothesis is amendable to further scientific validation from biomarkers with end-point sensitivity to oxygen radicals. So far, the biomarker programme developed around this theme may be divided into three distinct classes: (1) Markers of genotoxic exposure estimate DNA damage either directly as a biologically effective dose, or indirectly by estimating aberrant cellular functions that lead to accumulation of DNA damage. The examples included are ADP-ribosylation in mononuclear leucocytes (R. Pero, A. Olsson), oxidative DNA damage (K. Frenkel), gene expression in lymphocytes (S. Garte, G. Cosma), serum alpha macroglobulin (W. Troll) and oxidized DNA damage and repair (N. Christie). (2) Markers of genetic predisposition have been shown to have genetic inheritance patterns that relate to individual susceptibility to cancer or cardiovascular disease. The examples included are glutathione transferase mu phenotyping (R. Pero, J. Seidegard) and poly (ADP-ribose) polymerase pseudogene polymorphism (M. Smulson). (3) Markers of dietary status have been validated to estimate the amount of a particular nutrient or xenobiotic in the diet that has been taken up and metabolized or distributed to body fluids or tissues. The example included here is niacin nutriture (E. Jacobson, M. Jacobson). This biomarker is presented in Section 5 (pp. 59-62) of the Malmo Diet and Cancer Programme Minisymposium reported in this issue of the journal. PMID- 8429291 TI - Patients with persistent elevation of aminotransferases: investigation with ultrasonography, radionuclide imaging and liver biopsy. AB - Eighty-three patients, submitted for investigation of the liver due to persistently elevated activities of serum aminotransferases were included in a study in order to compare the relative merits of liver biopsy, ultrasound and radionuclide imaging. From the liver biopsy it was found that 45 patients had fatty liver, 14 had cirrhosis and 11 had chronic inflammation, 3 had haemochromatosis and 10 had unspecific changes or normal findings. An investigation with ultrasound yielded pathological findings in 65% of the patients. The sensitivity was highest in patients with fatty liver (82%) and more than 10% fat in the liver always resulted in increased echogenicity. Alterations in the liver due to cirrhosis and chronic inflammation were detected with ultrasound in only 50% and 57% of the cases, respectively. Radionuclide imaging was positive in 44% of the patients with fatty liver. 64% of those with cirrhosis and 21% of the patients with chronic inflammation. It was only possible to distinguish fatty liver from cirrhosis in those patients who had an increase in the size of the spleen (four patients). The study demonstrates that in patients with a persistent elevation of serum aminotransferases ultrasound has a high sensitivity for detecting more than 10% fat in the liver. However, both the ultrasound and a liver scintigram had a rather low sensitivity and a very low specificity for making a diagnosis for this group of patients. With the aid of a liver biopsy it was possible to establish a diagnosis in 90% of the patients. PMID- 8429292 TI - An epidemiologic perspective on biomarkers. AB - The authors discuss biological markers from an epidemiologic perspective, emphasizing the importance of integrating biomarkers into large-scale observational and intervention studies. Whereas any biologic phenomenon can be considered a biomarker, an intermediate end-point is defined as being on the causal pathway between exposure and disease. An intermediate end-point is a valid surrogate for a disease in relation to a given exposure if, and only if, that exposure causes a similar change in the occurrence of both the intermediate end point and the disease. Cancer studies using surrogate end-points may be shorter, smaller and cheaper than those using malignancy per se as an outcome. Three types of studies may be carried out to determine whether a given biomarker is an intermediate end-point and whether it can serve as a surrogate: (i) exposure marker studies, (ii) marker-disease studies, and (iii) studies comprising all three elements, exposure, marker, and disease. The authors discuss statistical aspects of these three types of studies and provide examples from investigations of alcohol-hormone-breast cancer, diet-epithelial proliferation markers-large bowel adenomatous polyps, and reproductive risk factors-human papillomavirus infection-cervical cancer. PMID- 8429293 TI - Response to antituberculous chemotherapy after splenectomy. AB - A case of a 17-year-old Indian male with disseminated tuberculosis is reported. While on chemotherapy, he was detected to have splenic abscesses. The patient remained unresponsive to antituberculous drugs until a splenectomy was performed. PMID- 8429294 TI - Azathioprine-associated pure red cell aplasia. AB - Azathioprine-associated pure red cell aplasia in patients with a renal allograft is a rare complication. Although immunological inhibition of erythroid progenitor cell has been suggested, the cause of this phenomenon remains unclear. The patient we describe showed a decrease in the number of erythroid progenitor cells and no evidence for the presence of a serum inhibitor of these precursor cells. Discontinuation of azathioprine was associated with a complete recovery from anaemia as well, with an increase in the number of erythroid progenitor cells. PMID- 8429296 TI - Clonidine in ulcerative colitis and proctitis. PMID- 8429295 TI - Syndrome of inappropriate antidiuretic hormone secretion (SIADH) and enalapril. AB - A 69-year-old woman with a history of diabetes and hypertension, was referred to the Hospital of Laredo because of hyponatraemia. She had weakness and slight dyspnoea with no evidence of extracellular fluid volume depletion or oedema. Serum sodium level on admission was 125 mol l-1, plasma osmolality 270 mosmol kg 1, simultaneous urine osmolality was 580 mosmol kg-1 and urine sodium 32.6 mmol l 1. She had been treated with enalapril (20 mg) daily for 4 months. She was diagnosed with the Syndrome of inappropriate secretion of antidiuretic hormone (SIADH) that was reversed after cessation of treatment with enalapril and reappeared on reintroduction of the drug at the same daily doses. PMID- 8429297 TI - Potentiation of acenocoumarol action by L-carnitine. PMID- 8429298 TI - Classification of tospoviruses based on phylogeny of nucleoprotein gene sequences. AB - The nucleotide sequences of the nucleoprotein (N) genes of seven tospovirus isolates representing three serogroups were determined and used to establish phylogenetic parameters to delineate species within the Tospovirus genus of the Bunyaviridae. A high sequence divergence (55.9% identity at the nucleotide level) was observed between isolates of serogroup I (tomato spotted wilt virus) and isolates of serogroup III (Impatiens necrotic spot virus). The serogroup II isolates take an intermediate position. Their N genes have 75% identity with those of serogroup I isolates and 57% with those of serogroup III isolates. Whereas the isolates within serogroups I or III have almost identical sequences, the two isolates BR-03 and SA-05 of serogroup II diverged significantly from each other (82.1% sequence identity). The results obtained support the conclusion that, in addition to the species TSWV and INSV, the serogroup II isolates BR-03 and SA-05 have to be considered as distinct species within the genus Tospovirus for which the names tomato chlorotic spot virus and groundnut ringspot virus, respectively, are proposed. PMID- 8429299 TI - Tomato necrosis and the 369 nucleotide Y satellite of cucumber mosaic virus: factors affecting satellite biological expression. AB - To determine which factors can affect biological expression of the Y satellite RNA of cucumber mosaic virus (CMV) in tomato, three laboratories collaboratively exchanged their natural satellite variants, the corresponding recombinant DNA clones and helper virus strains, as well as tomato varieties, on which different observations previously reported were based. The effects of these materials and the influence of temperature on symptom expression were systematically studied. The results show that in a standardized tomato bioassay at 24 degrees C, the Y satellite, when supported by either CMV-1 or CMV-Y, did not induce tomato necrosis in the Rutgers variety but elicited a slower necrotic response in the Best of All variety that was variably lethal, as compared to the faster inevitably lethal response induced by a prototype necrogenic D satellite variant in both tomato varieties. At higher temperatures (26.5 to 32 degrees C) an extremely fast-killing necrosis caused by CMV-Y itself was observed. The study demonstrates that in experiments on virus symptom modulation induced by CMV satellites, the nature of the helper virus, host plant varieties, as well as the environmental conditions should be precisely defined, and the effects of each parameter change determined separately. PMID- 8429300 TI - Biologically active transcripts from cloned cDNA of genomic grapevine fanleaf nepovirus RNAs. AB - Transcripts were produced in vitro by run-off transcription from full-length cDNA of RNA1 and RNA2 of grapevine fanleaf nepovirus (GFLV; isolate F13) cloned downstream from a bacteriophage RNA polymerase promoter. These transcripts, which possess a 5' terminal cap structure and a non-viral G residue instead of the naturally occurring genome-linked viral protein (VPg), are infectious to Chenopodium quinoa protoplasts when inoculated by electroporation. Synthetic RNA1 alone replicated in protoplasts. Inoculation of C. quinoa plants with synthetic RNA1 plus RNA2 produced symptoms similar to, but weaker, than those observed in plants infected with natural GFLV 6 to 8 days post-inoculation. Co-inoculated RNA1 and RNA2 were able to replicate and spread systemically in plants but RNA1 alone produced no symptoms and was not detected in non-inoculated leaves, suggesting that virus spread requires RNA2. Analysis of the genomic RNAs in plants infected with transcripts showed that the non-viral G at their 5' ends was not retained in the progeny. PMID- 8429301 TI - Processing of the dengue virus type 2 proteins prM and C-prM. AB - A glycoprotein C-prM of 35,000 M(r) was immuno-precipitated from lysates of Aedes albopictus cells infected with dengue virus type 2 (DEN-2) using antisera directed against the C protein or an amino-terminal fragment of the prM glycoprotein. C-prM was not detected in infected Vero cells. The prM glycoprotein synthesized in infected A. albopictus and Vero cells was cleaved to produce the membrane-associated virion protein (M) and the non-M fragment (pr) immediately preceding or occurring simultaneously with the release of viral particles from cells. The cleavage was less efficient in mosquito cells. The pr fragment was found only in the medium and was not rapidly degraded. To obtain pr-specific and M-specific antisera for these studies, proteins containing fragments of DEN-2 prM fused with staphylococcal Protein A were synthesized in Escherichia coli using the expression vector pRIT2T. The fusion proteins were stable and were used to raise antisera in rabbits for immunoprecipitation of radiolabelled cell extracts and culture medium. This is the first report of the detection of a C-prM protein in flavivirus-infected cells and the identification of the pr component of prM. PMID- 8429302 TI - Expression and immunogenicity of the entire human T cell leukaemia virus type I envelope protein produced in a baculovirus system. AB - The entire envelope gene of human T cell leukaemia virus type I (HTLV-I) has been successfully expressed in a baculovirus non-fusion vector system. The HTLV-I envelope protein accumulated within the insect cells as inclusion bodies which allowed efficient recovery of the recombinant protein. In an attempt to study the role of the HTLV-I envelope glycoprotein as an immunogenic target, mice were immunized with the envelope protein inclusion bodies (env-I.B.) in the presence or absence of an adjuvant. Antibodies of broad specificity were produced against the HTLV-I envelope protein in the presence or absence of an adjuvant as detected by Western blotting, radioimmunoprecipitation and peptide ELISA. Neutralizing antibody was detected when env-I.B. immunizations were carried out in the presence of high doses of a new adjuvant composed of a mycobacterial cell wall extract. In a combined immunization regimen, env-I.B. were found to enhance and broaden the antibody response to the HTLV-I envelope glycoprotein, following priming with various recombinant vaccinia virus (RVV) constructs expressing either the entire native HTLV-I envelope (gp46 and gp21) or just the surface envelope protein (gp46). Increased titres of neutralizing antibodies were observed following priming with the RVV expressing gp46 only. Results indicate that immunization regimens that involve priming with RVV expressing HTLV-I envelope followed by boosting with recombinant baculoviral HTLV-I envelope might be useful in eliciting protective immune responses in vivo. PMID- 8429303 TI - The distribution of Sindbis virus proteins in mosquito cells as determined by immunofluorescence and immunoelectron microscopy. AB - Two Aedes albopictus (mosquito) subclones, C7-10 and C6/36, were examined by immunofluorescence and immunoelectron microscopy for the distribution of Sindbis virus structural and non-structural proteins. Both the viral glycoproteins, E1 and E2, and the non-structural proteins, nsP1 and nsP2, were found within vesicles and electron-dense, amorphous matrices associated with Sindbis virus infection. The labelling patterns indicated that both replication of viral RNA and production of virus particles were localized within the same structures in the infected cell. The data support previous reports that alphavirus infection is contained within specific structures in the cytoplasm and provide additional evidence that the C6/36 and C7-10 subclones may represent different tissue types in the adult insect. PMID- 8429304 TI - Cloning and sequence analysis of the phosphoprotein gene of rinderpest virus. AB - We have cloned several cDNAs derived from the P gene of rinderpest virus. One of these, derived from a bicistronic N-P mRNA, has been sequenced in its entirety. Sequencing of a section of the others, and comparison with the genome sequence, showed that P gene transcripts, as for other morbilliviruses, were variable; non templated Gs could be added at a site resembling the normal stop transcription site. Primer extension analysis showed that about half the transcripts were edited. Sequences of the P, C and V proteins encoded by the normal and edited transcripts were compared with those of other morbilliviruses and with those of the more distantly related paramyxoviruses. PMID- 8429305 TI - The growth of cell culture-attenuated rinderpest virus in bovine lymphoblasts with B cell, CD4+ and CD8+ alpha/beta T cell and gamma/delta T cell phenotypes. AB - Cloned bovine lymphoblastoid cell lines, transformed by the protozoan parasite Theileria parva were infected with cell culture-attenuated rinderpest virus vaccine. The virus grew readily in lymphoid B cells, CD4+ and CD8+ alpha/beta T cells and gamma/delta T cells producing new infectivity, viral antigens, c.p.e. and total cell death. There did not appear to be a predilection for any particular phenotype of lymphoblast. The results imply that if the vaccine causes immunosuppression, it could do so through a variety of mechanisms. PMID- 8429306 TI - Importance of conserved amino acids at the cleavage site of the haemagglutinin of a virulent avian influenza A virus. AB - The virulence of avian influenza A viruses depends on the cleavability of the haemagglutinin (HA) by an intracellular protease at multiple basic amino acids. Although previous studies have demonstrated the importance of these amino acids for processing by the cellular protease, with emphasis on conserved residues near the cleavage site, the minimal requirements for cleavage remain unknown. By expressing site-specific mutants of the HA of a virulent avian influenza virus, A/turkey/Ireland/1378/85 (H5N8), in the simian virus 40 system and testing for their cleavability by an endogenous protease in CV-1 cells, and their fusion activity in a polykaryon formation assay, we were able to show that glycine at the amino terminus of HA2 is not essential for cleavage and that maximal cleavage requires at least five basic residues at the cleavage site, when carbohydrate is nearby. Moreover, we confirmed, that a conserved proline upstream of the cleavage site is not essential for HA cleavage or fusion activity, and that lysine replacement of the carboxyl-terminal arginine of HA1 abolishes cleavability. These findings should help identify the proteases responsible for intracellular cleavage of the HA of virulent avian influenza viruses. PMID- 8429307 TI - Molecular cloning and sequence analysis of a segment from Andean potato mottle virus B RNA encoding the putative RNA polymerase. AB - Andean potato mottle virus (APMV), an endemic South American comovirus, has a bipartite genome consisting of two plus-strand RNA molecules (M and B RNA). We have cloned the 3' half of the B RNA and identified the complete sequence of the putative APMV RNA polymerase. The RNA polymerase gene is part of a large polyprotein-encoding open reading frame. The putative, mature RNA polymerase, as deduced by comparison with the related cowpea mosaic virus (CPMV), type member of the comovirus group, is 703 amino acids long and shows a large degree of similarity with CPMV and other RNA polymerases. PMID- 8429308 TI - Protection against cucumber mosaic virus (CMV) strains O and Y and chrysanthemum mild mottle virus in transgenic tobacco plants expressing CMV-O coat protein. AB - Transgenic tobacco expressing the coat protein (CP) of cucumber mosaic virus strain O (CMV-O) showed a significant level of protection against CMV strains O and Y. When inoculum concentrations were increased, the transformants showed a stronger level of protection against CMV-O than against CMV-Y. The substitution of an amino acid residue between CMV-O and CMV-Y, which is presumed to change the conformation of CP, may reflect the difference in susceptibility to these viruses. Furthermore the transgenic tobacco plants showed a significant level of protection against chrysanthemum mild mottle virus, a member of the cucumovirus group but with no serological relationship to CMV. PMID- 8429310 TI - Temporal summation--the key to motor evoked potential spinal cord monitoring in humans. AB - Spinal motor evoked potentials (SMEP) were recorded from tibialis anterior muscle after epidural stimulation of the spinal cord at the low cervical or high thoracic level during scoliosis surgery. By using a double stimulus pulse to produce temporal summation within the spinal cord a maximal CMAP response was readily achieved despite good surgical anaesthesia. PMID- 8429309 TI - Apomorphine in treatment of Parkinson's disease: comparison between subcutaneous and sublingual routes. AB - The efficacy of two routes of apomorphine, subcutaneous (SC) and sublingual (SL), successively administered in 7 Parkinsonian patients with motor fluctuations, was compared in reducing the daily duration of "off" phases. The mean duration of SC and SL treatment was 7.7 and 6.8 months respectively. The mean time spent in "off" phase was 55% after SC and 68% after SL treatment. The mean time before turning "on" after an "off" period was 14 minutes after SC and 28 minutes after SL treatment. Two patients developed stomatitis after SL route. SL apomorphine may be helpful in the treatment of motor fluctuations in PD. PMID- 8429311 TI - Hereditary neuralgic amyotrophy associated with a relapsing multifocal sensory neuropathy. AB - A family with neuralgic amyotrophy (idiopathic brachial plexus neuropathy) associated with a multifocal sensory neuropathy is described. Four members over two generations were affected by neuralgic amyotrophy, inherited as an apparent autosomal dominant trait; two also had a multifocal relapsing sensory neuropathy with the clinical features of Wartenberg's migrant neuropathy. PMID- 8429312 TI - Calcified miliary brain metastases with mitochondrial inclusion bodies. AB - A patient with calcified miliary brain metastases from lung adenocarcinoma is reported. Electron microscopic study of the metastatic tumour cells showed membranous inclusion bodies in mitochondria. PMID- 8429313 TI - Early diagnosis and intravenous immune globulin therapy in paraneoplastic cerebellar degeneration. PMID- 8429314 TI - Absence of the amyloid precursor protein gene mutation (APP717: Val->Ile) in 85 cases of early onset Alzheimer's disease. PMID- 8429315 TI - Palatopharyngolaryngeal myokymia resembling "palatal myoclonus". PMID- 8429316 TI - Balint's syndrome in subacute HIV encephalitis. PMID- 8429317 TI - Anti-acetylcholine receptor antibody measurement in myasthenia gravis. PMID- 8429318 TI - Antithrombotic therapy in acute ischaemic stroke: an overview of the completed randomised trials. AB - A formal statistical overview of all truly randomised trials was undertaken to determine whether antithrombotic therapy is effective and safe in the early treatment of patients with acute stroke. There were 15 completed randomised controlled trials of the value of early antithrombotic treatment in patients with acute stroke. The regimes tested in acute presumed or confirmed ischaemic stroke were: heparin, 10 trials with 1047 patients: oral anticoagulants, one trial with 51 patients: antiplatelet therapy, three trials with 103 patients. Heparin was tested in one trial with 46 patients with acute haemorrhagic stroke. Outcome measures were deep venous thrombosis (confirmed by I125 scanning or venography), pulmonary embolism, death from all causes, haemorrhagic transformation of cerebral infarction, level of disability in survivors. In patients with acute ischaemic stroke, allocation to heparin was associated with a highly significant 81% (SD 8, 2p < 0.00001) reduction in deep venous thrombosis detected by I125 fibrinogen scanning or venogram. Only three trials systematically identified pulmonary emboli, which occurred in 6/106 (5.7%) allocated control vs 3/132 (2.3%) allocated heparin, a non-significant 58% reduction (SD 45.7, 2p > 0.1). There were relatively few deaths in the trials in patients with presumed ischaemic stroke: 94/485 (19.4%) among patients allocated to the control group vs 79/497 (15.9%) among patients who were allocated heparin. The observed 18% (SD 16) reduction in the odds of death was not statistically significant. The least biased estimated of the effect of treatment on haemorrhagic transformation of the cerebral infarct (HTI) comes from trials where all patients were scanned at the end of treatment, irrespective of clinical deterioration; using this analysis, haemorrhagic transformation occurred in 7/102 (6.9%) control vs 8/106 (7.5%) treated, a non-significant 12% increase (SD 56, 2p > 0.1). These data cannot exclude the possibility that heparin substantially increases the risks of HTI. No data on disability in survivors could be obtained. Early heparin treatment might be associated with substantial reductions in deep venous thrombosis (and probably also pulmonary embolism) and possibly a one fifth reduction in mortality (equivalent to the avoidance of 20-40 early deaths per thousand patients treated.) However, the data were wholly inadequate on safety, particularly on the risk of haemorrhagic transformation of the infarct and on the hazards of heparin therapy in patients with known intracerebral haemorrhage. The trials of oral anticoagulants (15 deaths among 57 patients) and antiplatelet therapy (two deaths among 103 patients) were too small to be informative. Much larger randomized trials-comparing aspirin, heparin and the combination of both drugs against control-in patients with acute ischaemic stroke are justified (and several are now planned or underway). PMID- 8429319 TI - Benign childhood partial epilepsies: benign childhood seizure susceptibility syndromes. PMID- 8429320 TI - Dystrophin analysis using a panel of anti-dystrophin antibodies in Duchenne and Becker muscular dystrophy. AB - Dystrophin, the protein product of the Duchenne muscular dystrophy (DMD) gene, was studied in 19 patients with Xp21 disorders and in 25 individuals with non Xp21 muscular dystrophy. Antibodies raised to seven different regions spanning most of the protein were used for immunocytochemistry. In all patients specific dystrophin staining anomalies were detected and correlated with clinical severity and also gene deletion. In patients with Becker muscular dystrophy (BMD) the anomalies detected ranged from inter- and intra-fibre variation in labelling intensity with the same antibody or several antibodies to general reduction in staining and discontinuous staining. In vitro evidence of abnormal dystrophin breakdown was observed reanalysing the muscle of patients, with BMD and not that of non-Xp21 dystrophies, after it has been stored for several months. A number of patients with DMD showed some staining but this did not represent a diagnostic problem. Based on the data presented, it was concluded that immunocytochemistry is a powerful technique in the prognostic diagnosis of Xp21 muscular dystrophies. PMID- 8429321 TI - Intravenous immunoglobulin treatment in patients with chronic inflammatory demyelinating polyneuropathy: a double blind, placebo controlled study. AB - Patients with a clinical diagnosis of chronic inflammatory demyelinating polyneuropathy (CIDP) were randomised in a double-blind, placebo-controlled multicentre trial to investigate whether high-dose intravenous immunoglobulin treatment (IVIg) for 5 consecutive days has a beneficial effect. Fifteen patients were randomised to IVIg and 13 to placebo. In the IVIg treatment group 4 patients improved and 3 patients in the placebo group. The degree of improvement of the patients in the IVIg treatment group was no different from the patients in the placebo group. Electrophysiological studies did not show significant differences between the groups. Since a previously performed cross-over trial showed that a selected group of CIDP patients responded better to IVIg than to placebo, it is concluded that we need better criteria to select CIDP patients for treatment with IVIg. PMID- 8429322 TI - Observations of improvement of reaching in five subjects with left hemiparesis. AB - Kinematic analysis and surface electromyography were used to study reaching by five subjects with left hemiparesis as they attempted to touch each of three targets. The targets were placed to require movement within and out of extensor synergy. Each subject was tested five times over a nine week period. Over this time, amplitude of peak velocity and sense of limb position significantly improved in the paretic arms. The increase of amplitude of peak velocity was more strongly related to a decrease in the discontinuity of movement (r = -0.48) than to maximal level of contraction of the prime movers (anterior deltoid: r = 0.25; biceps: r = 0.39). This finding may be a sign of learning or increased maturity of reach. These results, if replicated in a larger sample, would support therapy designed to improve learning of new sensorimotor relationships in the hemiparetic limb. PMID- 8429323 TI - Benjamin Franklin (1706-90). PMID- 8429325 TI - Age influences magnitude but not duration of response to levodopa. AB - Following an all-night fast, 45 patients with Parkinson's disease were examined using certain motor items present in the United Parkinson's Disease Rating Scale. All were given a single tablet of carbidopa 25 mg and levodopa 250 mg and re examined 90 minutes later. In addition to this evaluation, 23 of these patients underwent further scoring over a 4-hour period. A significant negative correlation was found between age and one important aspect of drug-derived benefit: magnitude of response. In contrast, age had no apparent influence on duration of benefit from the drug. Although baseline (fasting) scores were predictably correlated with duration of disease, magnitude of response was not adversely influenced by this variable. Not all Parkinsonian signs were equally influenced by age. Whereas the poor response of gait and bradykinesia appeared to be dependent on age, no such effect was noted on rest tremor scores. The data indicate that in patients with Parkinson's disease treated long term, factors associated with age rather than duration of disease may have a stronger adverse influence on magnitude of response to levodopa. PMID- 8429324 TI - Current management of ischaemic stroke. PMID- 8429326 TI - Intradural conus and cauda equina tumours: a retrospective review of presentation, diagnosis and early outcome. AB - This is a retrospective review of the clinical presentation, diagnosis and management of 62 patients with histologically proven intradural conus and cauda equina tumours. In the majority of cases the clinical presentation clearly suggests the need for further investigation. One fifth of the patients had small intramedullary tumours, which presented particular diagnostic difficulty and required sophisticated cross sectional imaging. PMID- 8429327 TI - Simultaneous recording of cerebrospinal fluid pressure and middle cerebral artery blood flow velocity in patients with suspected symptomatic normal pressure hydrocephalus. AB - CSF pressure (intracranial pressure, in one patient lumbar pressure) was monitored continuously for one night in 23 patients with suspected symptomatic normal pressure hydrocephalus (NPH) to identify patients who might benefit from subsequent shunt surgery. In 20 patients middle cerebral artery (MCA) blood flow velocity by means of transcranial Doppler sonography (TCD) and CSF pressure were recorded simultaneously. In three patients transcranial Doppler signals were insufficient. Spontaneous changes in CSF pressure always paralleled changes in the TCD signal. Equivalents of B-waves as well as intermediate waves (in between B- and A-waves), and C-waves could be identified easily and always appeared in phase. The Doppler signal, however, could not be used to evaluate the absolute changes in CSF pressure. Fast Fourier Transform of the Doppler signal was a useful tool to indicate the relative frequency of B-wave equivalents. In five patients the injection of 10ml saline into the ventricle raised intracranial pressure considerably, but hardly affected the MCA blood flow velocity. Continuous TCD monitoring might be useful as a noninvasive screening procedure in patients with suspected symptomatic NPH before continuous invasive CSF pressure measurements are performed. PMID- 8429328 TI - Severe psychiatric disturbance and abnormalities of the corpus callosum: review and case series. AB - The association between developmental defects of the corpus callosum and major psychiatric disturbance is discussed with a review of published cases. Seven new cases are presented, of which four had clear psychotic symptoms, two receiving a diagnosis of schizophrenia. Of the remainder, one had a developmental disorder affecting social interaction and speech which could be classed as Asperger's syndrome, one had a personality disorder with depressive and conversion symptoms, and the last was an adolescent boy with severe behavioural problems. The difficulties in determining the precise relevance of the callosal anomalies to these clinical manifestations are discussed especially since the prevalence of such anomalies in the population is uncertain. PMID- 8429329 TI - Behavioural treatment of slow cortical potentials in intractable epilepsy: neuropsychological predictors of outcome. AB - The study aimed to explore the predictive value of neuropsychological tests within the context of acquisition of slow cortical potential (SCP) self-control, a technique which has beneficial effects on seizure frequency in epilepsy. Patients with epilepsy who successfully achieved SCP control had longer digit or block-tapping spans than less successful patients. Patients who showed a better learning rate across training also displayed better verbal memory and learning abilities. Seizure reduction was related to block-tapping spans only. The results indicate that measures of attention, as indicated by digit spans or block-tapping spans, offer some predictive value for acquisition of SCP control and treatment outcome, whilst measures of visuospatial or frontal lobe function are unrelated to SCP acquisition and seizure reduction. PMID- 8429330 TI - Duration of illness in Huntington's disease is not related to age at onset. AB - The age at onset and duration of illness were studied in patients with Huntington's disease in the Leiden Roster which at 1 July 1990 contained 2787 patients. Of 1106 patients, 800 deceased and 306 alive, the age at onset was known. The median duration was 16.2 (range 2-45) years. In contrast to the current opinion, the median duration was independent of the age of onset. The median duration in juvenile Huntington's disease was 17.1 years, which is much longer than reported in the literature, and comparable with the categories for the age of onset of 20-34 and 35-49 years. Only in the group where onset was over 50 years of age was the median duration somewhat shorter (15.6 years), which can be ascribed to unrelated causes of death. As age of onset and duration of illness are not related, at least two mechanisms to determine the clinical course have to be postulated: one for age of onset and another for duration of illness. Duration was shorter for males, especially for those with an affected father. PMID- 8429331 TI - Launching "molecular nuclear medicine" grants, DOE scales back funding for medical applications programs. PMID- 8429332 TI - Scientific highlights of the EANM Congress 1992. PMID- 8429333 TI - Can we agree on the best imaging procedure(s) for localization of pheochromocytomas? PMID- 8429334 TI - Splenectomy in idiopathic thrombocytopenic purpura: its correlation with the sequestration of autologous indium-111-labeled platelets. AB - We present a retrospective analysis of 111In-platelet sequestration studies in 111 patients with the clinical diagnosis of idiopathic thrombocytopenic purpura (ITP). Fifty-one of these patients underwent splenectomy, independent of the results of the 111In-platelet studies to determine if these isotopic results could accurately predict a beneficial response to splenectomy. Between January 1984 and June 1990, 111 patients who presented with ITP were subjected to a study of autologous 111In-labeled platelets through autotransfusion. The platelet sequestration site was splenic (81%), mixed (12%), or hepatic (7%). Fifty-one patients with persistent drug-resistant thrombocytopenia underwent splenectomy regardless of the isotopic results: 33 patients beyond 6 mo after diagnosis and 18 with high hemorrhagic risks before this delay. The follow-up median duration was 2.9 yr. Thirty-three of the 38 patients with splenic sequestration showed a normalized platelet count, as opposed to 2 of the 13 with mixed or hepatic sequestration (p < 0.001). In addition, platelet survival extended beyond 8 days in six patients, with no apparent sequestration site. The platelet isotopic study performed with this technique appears to be indicated in ITP: it guides clinicians in their decision to perform splenectomy and relates to a more central mechanism certain thrombocytopenias that are inappropriately categorized as ITP. PMID- 8429335 TI - Assessment of myocardial perfusion defect size after early and delayed SPECT imaging with technetium-99m-hexakis 2-methoxyisobutyl isonitrile after stress. AB - It is thought that the distribution of 99mTc-sestamibi undergoes minimal change during the first 4 hr after injection. Thus [99mTc] sestamibi unlike 201Tl should not demonstrate significant redistribution in ischemic myocardium. We tested this assumption by quantifying perfusion defect size in early and delayed clinical SPECT images obtained after exercise stress or after dipyridamole infusion using an automated algorithm. Twenty patients with coronary artery disease aged 55 +/- 10 yr (13 male and 7 female) underwent stress imaging. Technetium-99m-sestamibi was injected at peak exercise stress in 12 patients and after an intravenous infusion of dipyridamole in 8 patients; SPECT images were obtained 1 and 4 hr later. All patients underwent rest imaging with a second dose of 99mTc-sestamibi 1-3 days after the stress studies. Total left ventricular mass and left ventricular defect mass were quantified with an automated algorithm previously validated in animal and patient studies. Estimates of total left ventricular mass from studies obtained 1 hr after stress (341 +/- 90 g) were comparable to values obtained after 4 hr (336 +/- 89 g), with a correlation of r = 0.97; p < 0.0001. The left ventricular defect size 1 hr after exercise or dipyridamole infusion (149 +/- 74 g) was similar to that observed after 4 hr (151 +/- 73 g). The two measurements of hypoperfusion with stress were highly correlated (r = 0.91; p < 0.0001) and were significantly larger than the defect size at rest (121 +/- 70 g). These observations support the conclusion that 99mTc-sestamibi does not redistribute significantly in ischemic myocardium between 1 and 4 hr after injection. PMID- 8429336 TI - Left ventricular cavity-to-myocardial count ratio: a new parameter for detecting resting left ventricular dysfunction directly from tomographic thallium perfusion scintigraphy. AB - Patients with reduced left ventricular function or aneurysms have cavities that appear dark on SPECT thallium scintigrams. We hypothesized that a quantitative index, which relates thallium activity in the left ventricular cavity to that in the myocardium (C/M ratio), could provide information on left ventricular function. A group of 80 patients who had both exercise SPECT thallium imaging and cardiac catheterization were studied. The C/M ratio was obtained from the short axis tomogram on both exercise and rest images. Counts in a 2 x 2 pixel region of interest in the left ventricular cavity were divided by the number of counts in the "hottest" area of the myocardium. Plotting the angiographically determined ejection fraction against the C/M exercise and rest ratios, we observed a linear correlation between ejection fraction and both C/M ratios, r = 0.65 for C/M exercise and r = 0.67 for C/M rest ratio (p < 0.00001). Using data from 12 normal cardiac catheterization patients, we established the lower limit of normal; 50% for ejection fraction and 0.40 for the C/M ratios. A C/M exercise ratio < or = 0.40 identified 26 of 31 patients with an ejection fraction < or = 50%. A C/M exercise ratio > 0.40 identified 39 of 49 patients with an ejection fraction > 50%. These calculations yielded a sensitivity of 83% and specificity of 78% for the C/M exercise ratio. A similar analysis for C/M rest ratio revealed sensitivity of 61% and specificity of 92%. The present study shows that an abnormal C/M ratio correctly distinguishes patients with abnormal from normal ejection fractions with an accuracy of 81%. The C/M ratio is easily obtained, requires minimal processing time and is highly reproducible. These attributes may enable this index to add supplementary information regarding left ventricular function in addition to perfusion from thallium imaging. PMID- 8429337 TI - Noninvasive, real-time monitoring of renal function: the ambulatory renal monitor. AB - The objective of this study was to develop a method for the noninvasive, continuous and real-time monitoring of renal function. A radiation detector attached to a miniature data logger was used to monitor the clearance of the glomerular filtration agent 99mTc-diethylenetriaminepentaacetic acid from the extracellular space. The rate constant (k) for this clearance showed an excellent correlation with simultaneous glomerular filtration rate (GFR) measurements performed with a standard 125I-iothalamate clearance technique in 50 patients. Moreover, the reproducibility of the k measurement for an individual or a population was superior to the GFR measurement performed with the standard clearance technique. The procedure was also used to monitor the renal function in patients at risk for acute renal failure during angiography or in the intensive care unit under noninvasive and near real-time conditions. The results show that the technique detects rapid changes in renal function with a resolution time of 5 min in patients with normal renal function and 15 min in patients with severely impaired renal function. Since the method is noninvasive, precise and provides a near real-time measurement of GFR, its use may lead to an improvement in the management of patients in situations in which a rapid measurement is the major concern. PMID- 8429338 TI - The dosimetry of iodine-123-labeled TISCH: a SPECT imaging agent for the D1 dopamine receptor. AB - TISCH is an iodinated D1 specific dopamine receptor antagonist that may be useful as a SPECT imaging agent. This report documents its pharmacological safety in animals and its radiation dosimetry in humans. The dose of radiation that 123I TISCH delivered to seven healthy subjects was estimated with the absorbed fraction technique. Conjugate images of the body were serially acquired for up to 24 hr after the administration of a known amount of activity. The count rates in the organs that could be visualized were measured on each image. These count rates were corrected for attenuation with 123I transmission scans. The doses to the other organs that did not take up enough activity to be visualized on the images were estimated with established models. The dosimetry was calculated for each subject individually before the results were averaged. Rapid biological washout minimizes the radiation exposure to most organs. The dose to the large bowel is limiting in healthy volunteers. The proximal colon receives about 0.67 rad/mCi (180 microGy/MBq) or about 5 rads for every 7.5 mCi of TISCH injected. This low radiation burden should make it feasible to study the D1 dopamine receptor in patients who have neuropsychiatric disorders before and after treatment. PMID- 8429339 TI - Hepatobiliary scintigraphy in a pediatric population: determination of hepatic extraction fraction by deconvolution analysis. AB - A study was performed to assess the feasibility of measuring the hepatic extraction fraction (HEF) and hepatic half clearance time (HCT) in pediatric patients with a variety of hepatobiliary diseases. There were 45 children categorized into four groups: normal 12; obstruction 9; hepatocellular disease (HCD) 16 and miscellaneous 8. In the normal patients, the mean HEF was 99% +/- 3.6% and the HCT was 23.6 +/- 7.7 min. In the two disease categories, hepatocellular disease and obstruction, there was a wide range of HEF 15%-84% and 25%-100%, respectively. This reflected the varying degrees of liver dysfunction and/or cholestasis. The average results of HEF for the HCD group was significantly lower than controls, and the greatest difference of the HCD group and other disease groups was with the miscellaneous group. This was not, however, different from the obstructive group. There was, however, a large overlap of results, and differentiation between the disease groups was not possible. HCT from the disease groups also showed a prolonged average clearance when compared to normals, although this was not significant when the p value was adjusted for multiple comparisons. The normal HCT was 23.6 +/- 7.7 min, whereas the hepatobiliary disease groups ranged from 20 to 714 min. Again, there was considerable overlap of results in the disease groups. These functional parameters are feasible and applicable in the pediatric population. PMID- 8429340 TI - Technetium-99m-tetrofosmin as a new radiopharmaceutical for myocardial perfusion imaging. AB - A new cationic complex, [99mTc(tetrofosmin)2O2]+, where tetrofosmin is the ether functionalized diphosphine ligand 1,2-bis[bis(2-ethoxyethyl)phosphino]ethane, has been synthesized and evaluated for potential use in myocardial perfusion imaging. The structure of the complex has been determined by x-ray crystallography of the 99Tc analog. In comparison with previously reported 99mTc complexes of alkyl phosphines, the tetrofosmin species shows substantially increased clearance from nontarget tissue, especially blood and liver. A freeze-dried kit formulation has been developed. The kit provides a product of high radiochemical purity up to 8 hr after reconstitution at room temperature. PMID- 8429341 TI - Potential gallium-68 tracers for imaging the heart with PET: evaluation of four gallium complexes with functionalized tripodal tris(salicylaldimine) ligands. AB - Gallium-67 and 68Ga complexes have been synthesized with tripodal hexadentate salicylaldimine ligands derived from 1,1,1-tris(salicylaldiminomethyl)ethane, sal3tame. The four ligands evaluated contained alkoxy substituents (n-BuO-, iso BuO-, sec-BuO-, and n-PrO-) on the terminal ethane carbon of the ligand backbone. In the case of the n-PrO-derivative, the tris(salicylaldimine) ligand was additionally substituted with methoxy groups in the 5-position of the aromatic rings. The 67Ga and 68Ga-complexes of these ligands were prepared by ligand exchange from 67Ga- or 68Ga-acetylacetonate in ethanol. The nonradioactive Ga[(sal)3tame-O-iso-Bu] complex was similarly prepared and shown by x-ray crystallography to exhibit the expected pseudo-octahedral N3O3(3-) coordination sphere about the Ga3+ center. These Ga-radiotracers are highly lipophilic, as demonstrated by their octanol/water partition coefficients. Log P values of 3.1, 3.1, 2.6, and 2.5 were found for the [(sal)3tame-O-iso-Bu], [(sal)3tame-O-n-Bu], [(sal)3tame-O-sec-Bu], and [(5-MeOsal)3tame-O-n-Pr] complexes, respectively. Following intravenous injection into rats, these complexes are rapidly cleared from the blood and exhibit significant myocardial uptake. At 1 min postinjection, 2.4%, 2.0%, 2.1% and 1.1% of the injected dose was found in the heart for the iso BuO, n-BuO, sec-BuO, and n-PrO complexes, respectively, dropping to 1.0%, 0.8%, 0.8%, and 0.7% at 5 min. The corresponding heart-to-blood ratios are quite high: 17 +/- 3, 14 +/- 2, 12 +/- 2 and 3.5 +/- 0.4 at 1 min and 14 +/- 4, 10 +/- 1, 10 +/- 1 and 3.2 +/- 0.1 at 5 min postinjection. High quality myocardial images were obtained with PET in a normal dog using data collected from 2 to 10 min following intravenous injection of 68Ga[(sal)3tame-O-iso-Bu]. PMID- 8429342 TI - Rapid infarct imaging with a technetium-99m-labeled antimyosin recombinant single chain Fv: evaluation in a canine model of acute myocardial infarction. AB - Studies of monoclonal antibody-based imaging agents show that blood clearance is inversely proportional to molecular size, i.e., Fab or Fab' > F(ab')2 > IgG. Indium-111-antimyosin Fab-DTPA is a highly specific and sensitive marker for myocardial necrosis. An improvement on current antibody diagnostic imaging may result from the use of smaller labeled fragments. We report the first in vivo targeting of acute myocardial infarction with a novel recombinant single-chain Fv (sFv) antimyosin protein. The sFv (MW = 27,594) is approximately one-half the size of the Fab and is comprised of the heavy and light chain variable regions from the myosin-specific murine monoclonal antibody R11D10 which were joined by a 15-amino-acid linker and expressed as a fusion protein (sFv) in E. coli. The binding affinity of the sFv for cardiac myosin was similar to the affinity observed for the Fab fragment. Technetium-99m labeling of the sFv was accomplished by the attachment of a cleavable, ester-linked bifunctional chelator (RP-1). Comparative studies in mice showed 99mTc-sFv-RP-1 cleared significantly faster (p < 0.001) than 99mTc-Fab'-RP-1 and 111In-Fab-DTPA antimyosin fragments. Furthermore, measurement of 99mTc-sFv-RP-1 blood clearance in a canine model of acute myocardial infarction gave a mean T1/2 of 0.54 +/- 0.13 hr versus 2.80 +/- 0.57 and 2.58 +/- 0.64 hr for Fab-DTPA and Fab'-RP-1 (p < 0.05), respectively. Despite its comparatively rapid clearance, 99mTc sFv-RP-1 had similar uptake in the infarct compared to the Fab'-RP-1. In addition, infarct visualization was more rapid with the sFv. Thus, these data demonstrate antimyosin sFv possesses characteristics necessary for rapid imaging of myocardial necrosis. PMID- 8429343 TI - Waste handlers and hospitals at odds over unregulated nuclear medicine waste. PMID- 8429344 TI - Uptake of technetium-99m-teboroxime in cultured myocardial cells: comparison with thallium-201 and technetium-99m-sestamibi. AB - The effects of metabolic inhibition on the uptake of 99mTc-teboroxime were assessed in cultured myocardial cells and compared with 201Tl and 99mTc sestamibi. Metabolic impairment was induced by cyanide (CN), a blocker of the mitochondrial respiratory chain, iodoacetate (IAA), an inhibitor of the glycolytic pathway, and ouabain, an inhibitor of Na(+)-K+ sarcolemmal ATPase. Cellular viability was appreciated by the trypan blue exclusion method. The effects of low temperature and of cellular death resulting from osmotic lysis were also assessed. Net cellular uptake of the radiotracers and the amount of proteins in the culture dishes were measured. All experiments were performed in parallel with control conditions and the results were expressed relatively to the control values. Teboroxime uptake was clearly decreased at low temperature (29.6% +/- 2.2% at 0 degree C, p < 0.001), while metabolic inhibition or osmotic lysis had no definite effect. The uptake of 201Tl and sestamibi was severely diminished in the presence of a mixture of 5 mM CN and 0.1 mM IAA, but 201Tl was less resistant than sestamibi (13.7% +/- 0.3% and 73.5% +/- 3.3%, respectively, after 1 hr of preincubation, p < 0.001 for both). Uptake of both tracers was very low in the presence of dead cells (12.1% +/- 1.3% for 201Tl and 4.1% +/- 0.2% for sestamibi, p < 0.001 for both). Ouabain had a detrimental effect only on 201Tl uptake at doses higher than 100 microM. Of these three currently available coronary blood flow imaging agents, teboroxime shows the lowest sensitivity to metabolic impairment. PMID- 8429345 TI - Rhenium-186-labeled monoclonal antibodies for radioimmunotherapy: preparation and evaluation. AB - Rhenium-186 has been determined to be a leading radionuclide for radioimmunotherapy. However, the use of 186Re has been limited due to the lack of a convenient and efficient method by which the radionuclide can be bound to monoclonal antibodies. We have developed a simple technique to label IgM, IgG, fragmented antibodies and tumor necrosis factor-alpha with 186Re. This technique uses ascorbic acid (AA) for controlled reduction of antibody disulfide groups to sulfhydryls and SnCl2 in citric acid for the reduction of 186ReO4-. The labeling yields as determined by instant thin-layer chromatography, molecular filtration and gel filtration were greater than 95% and the colloid formation was less than 5%. The labeled antibodies were stable when challenged with 100 and 250 molar excess of DTPA and HSA for 24 hr at 37 degrees C. SDS-PAGE analysis and autoradiography of labeled IgM, IgG and F(ab')2 monoclonal antibodies indicated uniform labeling and that no fragmentation of the monoclonal antibodies had taken place during the labeling procedure. Immunospecificity of 186Re-labeled human neutrophil specific IgM, as determined by in vitro antigen excess assay, was comparable to that of indium-111-labeled c-DTPA-IgM and technetium-99m-labeled IgM. A nuclear histone specific 186Re-TNT-1-F(ab')2 was evaluated in mice bearing experimental tumors. The tumor/muscle ratios at 4 and 24 hr were 5.9 +/- 0.21 and 13.8 +/- 6.7, respectively compared to that of 2.4 +/- 0.3 at 4 hr p.i. with a nonspecific protein. The labeling technique is simple, reliable and has already been adapted to a single-vial kit preparation. PMID- 8429346 TI - Formulation of technetium-99m-aerosol colloid with improved delivery efficiency for lung ventilation imaging. AB - Static imaging of lung ventilation in multiple views or with SPECT requires a 99mTc aerosol which does not wash out during imaging. Commercially available 99mTc colloids have suitable physical properties but are inefficiently delivered from jet nebulizers. An in vitro system was used to compare the delivery of various 99mTc radiopharmaceuticals from nebulizer solutions. Delivery of noncolloidal agents (pertechnetate, pentetate, imidodiphosphate, albumin) was much greater than that of colloids. However, addition of phosphate buffer to an in-house 99mTc-tin colloid produced an agent that was delivered as efficiently as the noncolloidal tracers and which shows promise for clinical use. PMID- 8429347 TI - Comparison of the distribution of radioiodinated-E-17 alpha-iodovinyl-11 beta methoxyestradiol and 2-iodo-1,1-bis(4-hydroxyphenyl)-phenylethylene estrogens in the immature female rat. AB - Current therapies for estrogen receptor-positive (ER+) cancers rely heavily on growth prevention by cytostatic agents rather than destruction of the cancer cells. In these studies, we compared the tissue distribution of two radioiodinated estrogens, one a triphenylethylene estrogen, IBHPE, the other steroidal, IVME2. The radioiodoestrogens were prepared using the halodestannylation reaction from the respective tributyltin precursors. The specificity of binding of these radioiodinated estrogens to the estrogen receptor (ER) was established by sucrose-density gradient analyses and their specific activities by comparison with the ER binding of 3H-estradiol of known specific activity. The time-dependent tissue distribution of the two radioiodoestrogens in immature female rats was studied to compare the relative uptake and specific retention of the two estrogens in ER target organs and assess their possible use for imaging ER positive tissues or for Auger electron-mediated ER-directed therapy. While the uterus showed only slightly poorer retention of the nonsteroidal estrogen (IBHPE) than the steroidal estrogen (IVME2), those target tissues that required blood-supplied ligand (e.g., vagina, pituitary) showed substantially higher uptake of the steroidal estrogen. IBHPE showed significantly higher blood levels at all time points. While the tissue-to-muscle ratios for IBHPE in the uterus and ovary were higher initially, the IVME2 showed higher tissue-to-muscle ratios, suggesting that the steroidal estrogen may be the more promising ligand for imaging purposes. However, IBHPE showed excellent uptake by peritoneal target tissues, with much lower concentrations in more distant target tissues (e.g., pituitary) so it might have distinct potential for therapy of intraperitoneal ER-containing cancers. PMID- 8429348 TI - Inhomogeneity of gated and ungated SPECT technetium-99m-sestamibi bull's-eyes in normal dogs: comparison with thallium-201. AB - By computer simulation, we have previously hypothesized, independent of the isotope imaged, that differences in view-to-view resolution and attenuation patterns predictably cause count density distortions in SPECT images. We tested the simulation predictions for both ECG-gated and ungated SPECT 99mTc-sestamibi and SPECT 201Tl myocardial perfusion images in normal dogs. In agreement with the predictions of the computer model, distortions in SPECT 99mTc-sestamibi myocardial perfusion images are virtually equivalent to SPECT 201Tl, dependent on the exact SPECT acquisition orbit and markedly different for a posterior 180 degrees acquisition arc compared to an anterior 180 degrees acquisition arc. Furthermore, ungated and gated SPECT 99mTc-sestamibi images show similar count inhomogeneities. These results suggest that little is to be gained from a 360 degrees acquisition with SPECT 99mTc-sestamibi, and that image distortions from gated or ungated SPECT 99mTc images with 180 degrees orbits will be similar to those in SPECT 201Tl images. PMID- 8429349 TI - Fluorine-18-fluorodeoxyglucose positron emission tomography in technetium-99m hydroxymethylenediphosphate negative bone tumors. AB - We have encountered two cases of bone tumors with high 18F-fluorodeoxyglucose (FDG) uptake and negative 99mTc-HMDP bone scintigraphy, including a patient with myeloma and a patient with a metastatic bone tumor from esophageal cancer. Bone scintigraphy with a 99mTc-phosphate complex reflects osteoblastic activity in the bone tissue surrounding the tumor, whereas the accumulation of FDG is associated with the metabolic activity of the tumor itself. An FDG-PET study can therefore be used as a complementary study for the detection and follow-up of bone tumors when a 99mTc-phosphate bone scintigram is negative. PMID- 8429350 TI - Cerebral blood flow and magnetic resonance imaging in locked-in syndrome. AB - The cerebral blood flow (CBF) of a patient suffering from locked-in syndrome (LiS) was examined before and after the onset using 99mTc hexamethylpropyleneamine oxime single-photon emission computed tomography (SPECT) and the intravenous 133Xe injection method. The mean CBF during the locked-in state was 32.2 ml/100 g/min, a 42% reduction from the asymptomatic stage. SPECT showed profound reductions of perfusion in the bilateral cerebral cortices, subcortical regions and in the cerebellum, with a less marked reduction in the frontal cortices. On Day 49, the patient showed some minimal voluntary return with a moderate increase in mean CBF of 40.2 ml/100 g/min. The relative CBF values in the cerebral cortices and subcortical regions were restored, but the bilateral cerebellar hypoperfusion remained unchanged. SPECT and CBF are useful for a better characterization of the brain pathophysiology in LiS. PMID- 8429351 TI - Technetium pertechnetate scintigraphy in an ileal duplication of the stomach and duodenum. AB - We report a patient with ileal duplication of the stomach and duodenum that presented with gastrointestinal bleeding. The scan appearance of the duplicated stomach in both shape and temporal course of activity bore a striking resemblance to the patient's homotopic stomach. PMID- 8429352 TI - Scintigraphic "doughnut sign" on skeletal imaging due to a hemangioendothelioma of bone. AB - A 61-yr-old patient was referred to our hospital for investigation of pain and tenderness in his left lower limb. Bone scan revealed multiple lesions of tibia and foot, several of which appeared as doughnut-like lesions and corresponded to lytic abnormalities on radiographs. Pathologic examination revealed multiple epithelioid hemangioendothelioma of bone. PMID- 8429353 TI - Results of surgery for portacaval shunt in a patient with situs inversus diagnosed and evaluated by per rectal portal scintigraphy. AB - A 70-yr-old man diagnosed with situs inversus was hospitalized because of hepatic encephalopathy. Per rectal portal scintigraphy showed portal hypertension and a region with high radionuclide activity in the right lateral region of the abdomen. In percutaneous transhepatic portograms, a giant portacaval shunt was seen in the region with high radionuclide activity. After the portacaval shunt was obstructed surgically, the hepatic encephalopathy disappeared. In per rectal portal scintigraphy done 2 wk after surgery, the pattern was normal and the region with high radionuclide activity had disappeared. PMID- 8429354 TI - A quantitative assessment of patient motion and its effect on myocardial perfusion SPECT images. AB - Patient motion during image acquisition is a frequent cause of SPECT perfusion image artifacts. We sought to determine the relationship between patient motion and the resultant image artifact. The effect of patient motion on 201Tl SPECT scintigrams was assessed with computer simulation to create 66 new image sets with artifactual vertical, horizontal and combined patient motion introduced over a broad range in six normal studies. Visual analysis of regional radioactivity in these simulated images, as well as quantitative analysis of the resultant polar coordinate display was performed. The presence and extent of "motion" artifacts varied with the number and location of the projection images affected, as well as the extent of their displacement. Although the extent of the defect varied with the frames affected, they were not necessarily more extensive when related to vertical displacement in the center of the orbit. The location of induced defects varied with direction of displacement and the location of frames affected. Vertical and horizontal motion created additive defects. Defect size grew with incremental vertical displacement but subsequently decreased with yet increasing displacement. Both the irregular, "lumpy" distribution of radioactivity, often with opposing "defects", as well as curvilinear extraventricular radioactivity, were visual clues suggesting SPECT defects related to motion artifact. A clinical case review revealed that approximately 25% of studies demonstrate such motion during acquisition but only 5% contribute to visible image deterioration. While detection is important, postacquisition attempts to correct such artifacts are incomplete and optimally, they must be prevented. PMID- 8429355 TI - Three-dimensional registration of cardiac positron emission tomography attenuation scans. AB - A method is described and tested which allows three-dimensional registration of two cardiac PET attenuation scans of the same subject acquired at different times. The alignment method is based on maximizing the correlation coefficient between the stacks of slices obtained during each of the two imaging sessions. The method appears accurate to better than 1 mm in x, y and z, and better than 1.5 degrees in the three angles of rotation. The method has been tested by using it to realign deliberately misaligned attenuation scans, and also by using realigned attenuation scans to correct fluorodeoxyglucose (FDG) emission scan data. The FDG emission data reconstructed with the realigned attenuation scans do not differ appreciably from the FDG data obtained using the original attenuation data. The method should be useful in realigning emission data (using the alignment coordinates predicted by realigning the attenuation scans), correcting for subject motion between scans (especially multiple intervention scans) and may allow a subject to leave the scanning table between scans during long imaging sessions. PMID- 8429356 TI - Automated detection of the intercommissural line for stereotactic localization of functional brain images. AB - A technique has been developed for automated detection of the intercommissural (AC-PC) line for positron emission tomography (PET). The AC-PC line is estimated from the location of four internal landmarks; the frontal and occipital poles, the inferior aspect of the anterior corpus callosum, and the subthalamic point. The landmarks are detected automatically in PET mid-sagittal slices by combining edge detection, interpolation and profile curve analysis techniques. The anatomical relationships between the true and estimated AC-PC lines from the landmarks was confirmed by analysis of magnetic resonance (MR) images. Accuracy of the automated estimation technique was assessed in co-registered PET and MR images, which showed minimal angular differences and displacements of the estimated from the true AC-PC lines. The automated detection of the AC-PC line in a PET study enables accurate stereotactic localization of functional signals without the need for additional anatomical imaging and provides a basis for objective and reproducible intersubject comparison. PMID- 8429357 TI - The channel ratio method of scatter correction for radionuclide image quantitation. AB - The accuracy of quantitation of radionuclide distributions in human tissue with the scintillation camera is decreased by attenuation and scatter of photons. If scatter correction is applied satisfactorily, narrow beam attenuation can be applied. In this article, a scatter correction technique, the channel ratio (CR) method, is introduced. The CR scatter correction method is proposed for quantitation of the radionuclide distribution in organs. The improvement in the geometrical resolution was measured and examples of clinical images are presented. In this method, the change in the ratio of counts from two symmetrical adjacent energy windows straddling the energy photopeak was used to eliminate the contribution of scattered photons during imaging with 99mTc. The theory and methods for the empirical affirmation are described. To apply the CR scatter correction method, two constants, the ratio of primary photons G and the ratio of scattered photons H in the same windows, were determined. Different sized sources in varying depths of water were imaged. When the source activities were quantified after scatter correction with the CR method, the measurements ranged from 96%-108% in comparison to the reference value in 100 mm water. The scatter fraction increased from 0.20 in 10 mm water to 1.44 in 200 mm water. The geometrical resolution expressed as full width at tenth maximum in 150 mm water improved by 30.4% and was restored to the value of the geometrical resolution in air. The CR scatter correction method is a simple method to correct for scatter in order to facilitate accurate quantitation of the radionuclide distribution during imaging with a scintillation camera. PMID- 8429358 TI - Automated region definition for cardiac nitrogen-13-ammonia PET imaging. AB - In combination with PET, the tracer 13N-ammonia can be employed for the noninvasive quantification of myocardial perfusion at rest and after pharmacological stress. The purpose of this study was to develop an analysis method for the quantification of regional myocardial blood flow in the clinical setting. The algorithm includes correction for patient motion, an automated definition of multiple regions and display of absolute flows in polar map format. The effects of partial volume and blood to tissue cross-contamination were accounted for by optimizing the radial position of regions to meet fundamental assumptions of the kinetic model. In order to correct for motion artifacts, the myocardial displacement was manually determined based on edge-enhanced images. The obtained results exhibit the capability of the presented algorithm to noninvasively assess regional myocardial perfusion in the clinical environment. PMID- 8429359 TI - Quality assurance of white blood cell labeling with a test based on adherence. AB - A new quality control assay was developed based on the premise that proper radioactive labeling should not affect the adherence characteristics of white blood cells to nylon fibers. Heparinized whole blood with trace amounts of radioactively labeled white blood cells was passed over nylon fiber columns and eluted in eight fractions. Percent radioactive adherence (%RA) and percent white blood cell adherence (%WBCA) were determined for each fraction. Regression lines (%RA versus %WBCA) were calculated for 9 samples labeled properly with 111In oxine and for 17 samples intentionally subjected to improper labeling. Properly labeled preparations had a median slope = 1.05 and an intercept = 1%. Improperly labeled preparations had significantly lower slopes and/or higher intercepts. By the use of +/- 2 s.d. ranges as indicators of proper labeling (slope of 0.71 1.74; intercept of -35%-37%), the test had 100% sensitivity and 94% specificity. We conclude that proper labeling with 111In-oxine preserves the adherence characteristics of white blood cells, that improper labeling may affect the binding strength of white blood cells (decrease in slope) and/or lead to formation of sticky cell subgroups (increased intercept) and that the quality control assay can objectively assess the impact of labeling on adherence. PMID- 8429360 TI - Disposal of small quantities of radioactive materials in local landfills: problems and solutions. PMID- 8429361 TI - Cellular basis for elevated gallium-67 uptake in rheumatoid lung patient. PMID- 8429362 TI - Ventilation-perfusion lung scans. PMID- 8429363 TI - Strictures on outpatient nuclear medicine therapy. PMID- 8429364 TI - Infant diet affects serum lipoprotein concentrations and cholesterol esterifying enzymes in baboons. AB - We characterized the preweaning differences in cholesterol metabolism between breast-fed and formula-fed baboons and determined if formulas with low and high polyunsaturated:saturated fatty acid (P:S) ratios simulated the effects of breast feeding. At birth, 45 infant baboons from three sires and 44 dams were assigned to breast-fed, low P:S formula or high P:S formula diet groups until weaning at 14 wk. From 4 to 14 wk breast-fed infants had higher serum cholesterol because of much higher HDL1- and HDL2-cholesterol concentrations but had lower HDL3 cholesterol than both formula-fed groups. LDL-cholesterol was higher in infants fed the low P:S fomula. Breast-fed infants had higher serum apolipoprotein E than the formula-fed groups, but diet did not affect apolipoprotein A-I or B concentrations. Breast-fed infants had higher hepatic acyl CoA cholesterol acyltransferase activity and lower plasma lecithin cholesterol acyltransferase activity. These enzyme activities were not different between infants fed low or high P:S formulas. Post-heparinized plasma lipoprotein lipase activity was greater in breast-fed infants than in those fed formula. These findings demonstrate that the P:S ratio of formulas has little effect on cholesterol metabolism during the preweaning period and suggest that factors other than fat composition account for the metabolic differences between breast feeding and commercial infant formula. PMID- 8429365 TI - Diet during lactation associated with infant behavior and caregiver-infant interaction in a semirural Egyptian village. AB - Potential processes through which nutritional and non-nutritional factors can relate to infant state and behavior and mother-infant interactions were examined in 41 mother-infant pairs from semirural Egyptian households. All infants were breast-fed, and breast milk was the main source of their nutrient intake. Median birth weight was close to reference median; however, most infants showed growth faltering when they were 3-6 mo of age. Among the infant behavioral and state variables, only drowsiness (a proxy for activity and alertness) was significantly associated with the nutritional and non-nutritional factors examined. Among these factors, mothers' intakes of animal source foods and certain B vitamins were the strongest predictors of drowsiness. The nature of the association between maternal diet and drowsiness, examined by multiple regression analysis, showed clearly that inadequate diet of the mother was the major risk factor. Alertness of infants was further compromised when there were several children in the households. The small, less vocal and less alert infants received less vocalization from their mothers. In this environment, infants of undernourished mothers may not receive the extra care and stimulation needed and are at risk for subsequent developmental disabilities. PMID- 8429366 TI - Nutrient supplements contribute to the dietary intake of middle- and older-aged adult residents of Beaver Dam, Wisconsin. AB - Intake of macronutrients and micronutrients was estimated from a diet history questionnaire in a population-based sample of persons (n = 2152), 43-86 y of age. Men and women 65 y of age or older consumed less protein, fat, cholesterol, niacin, riboflavin, calcium, iron, zinc and lycopene than their middle-aged counterparts. After including nutrient intake from supplements, age differences in intake were no longer significant for zinc in women or for niacin and iron in men. After further adjusting for energy intake, only protein, cholesterol, and lycopene mean intakes were lower in older compared with middle-aged men and women. Regular use of supplements was most prevalent among older men and women (40 and 48%, respectively). Supplement use was greater in the current time period compared with retrospective reports of supplement use 10 y before interview. The impact of supplement intake on median values of intake was greatest in women, particularly for vitamins A, C and E, riboflavin and calcium. The high prevalence of supplement use emphasizes the importance of considering this source of nutrients in future investigations in which dietary intake is measured. PMID- 8429367 TI - Gender bias in food intake favors male preschool Guatemalan children. AB - Gender bias in food intake and its subsequent effects on growth and illness were examined using data from rural Guatemalan children. Multiple regression controlled for energy requirements, illness, and maternal and economic factors. Gender bias in energy and protein intake favored boys; the magnitude for ages 2-5 y was 247 kJ/d. Analysis of subsequent effects showed that boys had higher rates of weight gain due to gender bias in energy intake than did girls for ages 1-2 y (0.27-0.97 kg/y), when there were no differences in illness rates due to gender bias in energy intake. For age 3-5 y, boys and girls did not differ in weight gain due to gender bias in energy intake. For ages 1-2 y for weight and stature, the growth rate for boys was faster than that of girls by 6-49% due to gender bias. This study provides evidence of gender bias in food intake in a Latin American population, but more work on the existence of and reasons for gender bias in food intake is needed before advocating that education or health programs should focus on this issue. PMID- 8429369 TI - Vitamin B-6 deficiency alters rat enterocyte calcium homeostasis but not duodenal transport. AB - Isolated enterocytes were used as differential transporting cells to examine calcium homeostasis in control and vitamin B-6-deficient rats. Kinetic analysis of calcium fluxes, as well as biochemical determinations, indicated that enterocytes from control animals had high concentrations of cytosol ionized calcium (318.5 +/- 22.4 nmol/L) and a large pool of exchangeable calcium (2.72 nmol/mg protein, or 86% of total cell calcium). Vitamin B-6 deficiency resulted in a 44% reduction in total cellular calcium (1.71 +/- 0.24 vs. 3.07 +/- 0.29 nmol/mg protein), a 69% reduction in total exchangeable calcium (0.85 vs. 2.72 nmol/mg protein) and a 56% reduction in cytosol ionized calcium concentration (141.4 +/- 13.5 vs. 318.5 +/- 22.4 nmol/L). Calcium fluxes between all cellular compartments were markedly diminished as a result of vitamin B-6 deficiency. However, vitamin B-6 deficiency did not affect the basic morphological or functional features of the enterocytes, such as cell viability, cell volume, membrane permeability and protein content. Moreover, intestinal calcium transport in vivo was not affected during vitamin B-6 deficiency, perhaps due to the greater paracellular ion movement compensating for the lower transcellular transport. PMID- 8429368 TI - Periconceptional vitamin profiles are not suitable for identifying women at risk for neural tube defects. AB - Folic acid and other vitamin deficiencies may play a role in the etiology of neural tube defects. The Medical Research Council Vitamin Study confirmed the beneficial effect of folic acid supplementation on the prevention of neural tube defects. However, the concentrations of vitamins other than folate were not a common feature of any of the former studies. We measured the concentrations of vitamin A, riboflavin, riboflavine-5'-monophosphate, flavine-adenine dinucleotide, vitamin B-6, vitamin B-12, vitamin C, vitamin E, folate and ferritin in the serum of women who had previously had a child with a neural tube defect and were planning a further pregnancy. Vitamin and folic acid supplements were supplied before conception to 44 high risk women before conception. Eighteen other high risk women not given supplements were the control group. We concluded that vitamin profiles do not form a suitable means for identifying women at risk for neural tube defects before pregnancy. This endorses the hypothesis that the beneficial effect of folic acid supplementation on the prevention of neural tube defects is possibly at least partly due to the fact that it overrides a relative folic acid shortage caused by a metabolic disorder. PMID- 8429370 TI - Vitamin E alters hepatic antioxidant enzymes in rats treated with dehydroepiandrosterone (DHEA). AB - The effects of vitamin E on hepatic antioxidant enzymes and plasma indicators of tissue damage were studied in rats treated with dehydroepiandrosterone (DHEA). Thirty-two male Sprague-Dawley rats were randomly allotted to one of four groups of eight rats each. Rats were treated with DHEA [100 mg/(kg body wt.d), i.p.], vitamin E (1 g/kg diet), or DHEA+vitamin E, or were untreated (controls) for 5 wk. Treatment with DHEA reduced (P < 0.05) weight gain, fat pad weight and carcass lipid concentration and increased carcass protein and ash concentration compared with control rats. The DHEA-treated rats had significantly lower concentrations of serum triglycerides and total cholesterol, yet greater amounts of liver lipid, than did control rats. Supplementation of DHEA-treated rats with vitamin E had no significant effect on weight gain, carcass composition or plasma metabolites compared with rats treated with DHEA alone. The rate of hepatic peroxisomal fatty acid oxidation in DHEA-treated rats was approximately 240% of that in control or vitamin E-supplemented rats. The specific activities of enzymes that defend against oxidative stress (e.g., glutathione reductase, glutathione transferase, catalase) or are indicators of tissue damage (e.g., alanine and aspartate aminotransferases) were all significantly higher in DHEA treated rats compared with control rats. Supplementation of DHEA-treated rats with vitamin E generally reduced these indices of oxidative stress compared with rats treated with DHEA alone, suggesting that vitamin E may have a protective effect against potential oxidative damage associated with DHEA treatment. PMID- 8429372 TI - 1992 AIN Symposium Proceedings. AIN Annual Meeting. Anaheim, California, April 5 9, 1992. PMID- 8429371 TI - Low methionine ingestion by rats extends life span. AB - Dietary energy restriction has been a widely used means of experimentally extending mammalian life span. We report here that lifelong reduction in the concentration of a single dietary component, the essential amino acid L methionine, from 0.86 to 0.17% of the diet results in a 30% longer life span of male Fischer 344 rats. Methionine restriction completely abolished growth, although food intake was actually greater on a body weight basis. Studies of energy consumption in early life indicated that the energy intake of 0.17% methionine-fed animals was near normal for animals of their size, although consumption per animal was below that of the much larger 0.86% methionine-fed rats. Increasing the energy intake of rats fed 0.17% methionine failed to increase their rate of growth, whereas restricting 0.85% methionine-fed rats to the food intake of 0.17% methionine-fed animals did not materially reduce growth, indicating that food restriction was not a factor in life span extension in these experiments. The biochemically well-defined pathways of methionine metabolism and utilization offer the potential for uncovering the precise mechanism(s) underlying this specific dietary restriction-related extension of life span. PMID- 8429373 TI - Historical overview of the safety of the food supply. Introduction. PMID- 8429374 TI - Science, law and society: the pursuit of food safety. PMID- 8429375 TI - Safe foods for infants--the regulation of milk, infant formula and other infant foods. PMID- 8429376 TI - Contribution of nutritional sciences to food safety: control of mycotoxins. PMID- 8429377 TI - The Food Protection Committee: a non-government organization. PMID- 8429378 TI - Avian bone metabolism: cell-mediated mineralization and localized regulatory factors. PMID- 8429379 TI - Involvement of cellular metabolism of calcium and phosphate in calcification of avian growth plate cartilage. AB - Past work established that matrix vesicles (MV) are primary initiators of extracellular mineral deposition in endochondral calcification. Reviewed here are studies on how direct cellular metabolism of Ca2+ and inorganic phosphate (Pi), and cellular interaction with the matrix, are involved in the formation of calcifiable MV. Presented is a working model of how chondrocytes in growth plate (GP) cartilage are envisioned to induce the formation of calcifiable MV. In part, this model is based on recent laser confocal imaging of living cartilage tissue sections with Indo-I AM, a fluorescent permeant Ca2+ probe. These studies indicate that GP chondrocytes actively acquire Ca2+, concentrate it to the cell periphery and exfoliate it as Ca(2+)-rich MV. Data from direct chemical analysis and 31P-NMR studies on freshly isolated cells show that GP chondrocytes are depleted of ATP and have elevated cytosolic Pi, a condition prerequisite to formation of Ca(2+)-acidic phospholipid (APL)-Pi complex-primed MV. Chondrocyte cell membrane processes from which MV arise have been found to be tightly linked to the cartilage-specific extracellular matrix collagens and proteoglycans. Annexins V and VI, APL-dependent Ca(2+)-binding proteins that form Ca2+ channels in chondrocytes and MV membranes, also bind to the matrix collagens and may serve as mechano-transducers in GP cartilage, gating Ca2+ entrance into the cells and MV. This interaction between the extracellular matrix and chondrocytes appears to facilitate Ca2+ loading of chondrocytes, formation of Ca2+ and Pi-primed MV and rapid induction of mineralization in GP cartilage. PMID- 8429380 TI - The role of the vitamin D endocrine system in avian bone biology. AB - The involvement of vitamin D and its endocrine system is essential, both for the process of bone development and growth, as well as bone remodeling. Important bone cells participating in those processes include the osteoblast (bone formation), the osteoclast (bone resorption) and the growth plate chondrocyte (longitudinal bone growth). The hormonally active form of vitamin D3, 1,25 dihydroxyvitamin D3 [1,25(OH)2D3], generates many of the biological responses attributed to the parent vitamin D3, including actions on osteoblasts and chondrocytes and the stimulation of the production of osteoclasts. 1,25(OH)2D3 is able to generate biological responses via both genomic and nongenomic pathways. This review provides a summary of this area. PMID- 8429381 TI - Local modulation of skeletal growth and bone modeling in poultry. AB - Skeletal growth and bone modeling in poultry are regulated by complex interactions between the animal's genetic potential and a host of systemic and localized factors (growth factors and cytokines) influencing bone biology. The objective of these interactions is to orchestrate the achievement of bone architecture that balances functionally appropriate morphology with the skeleton's involvement in mineral homeostasis. Within this context, bone modeling in the growing animal represents an adaptive process that is distinct from bone remodeling, which is the term used to describe the resorption and formation of mineralized tissue that maintains skeletal mass and morphology in the adult. As many of the skeletal lesions that afflict poultry are the consequence of abnormalities in bone modeling, not bone remodeling, an appreciation of the differences between these two contrasting processes is a prerequisite for understanding the pathogenesis of skeletal lesions in poultry. PMID- 8429382 TI - Removal of glycylglutamine from plasma by individual tissues: mechanism and impact on amino acid fluxes in postabsorption and starvation. AB - A possible source of glutamine, for inclusion in the parenteral solutions, is glycylglutamine. The aim of this article is to review briefly the information on metabolism of glycylglutamine when administered intravenously. The fact that there is efficient utilization of intravenously infused glycylglutamine was evident with very little excretion in the urine. Although all the tissues examined, except brain, participated in the removal of glycylglutamine from plasma, kidney predominated in this regard. This may be related to the presence of carrier-mediated systems for cellular uptake of glycylglutamine in the kidney and the lack of them in other tissues. Starvation did not alter the metabolic clearance of glycylglutamine, although it reduced the removal by the kidney. Renal metabolism of glycylglutamine resulted in the release of constituent amino acids that were largely utilized by the liver in the postabsorptive state and by skeletal muscle in starvation. This alteration was accompanied by a selective inhibition of muscle release of amino acids that are substrates for enhanced hepatic gluconeogenesis and renal ammoniagenesis in starvation. Because there was no change either in plasma glucose level or ammonia excretion during the infusion of glycylglutamine in starved human subjects, apparently the amino acid residues of glycylglutamine fulfilled the substrate needs for these functions. These results provide a metabolic basis for further investigations of the possible nutritional benefit of including glycylglutamine in parenteral nutrition. PMID- 8429383 TI - Role of protein synthesis in amino acid catabolism. AB - Except for branched chain amino acids, the site of indispensable amino acid degradation is the liver. Location of amino acid degradation capacity in a single organ may play an important role in the reutilization of amino acids derived from protein turnover. The importance of preferential utilization of amino acids for protein synthesis on catabolism of amino acids is demonstrated in two ways. First, by minimal oxidation of an amino acid at dietary concentrations below that required for maximum gain followed by a near proportionate oxidation with increased dietary level, and second, by increased oxidation of an indispensable amino acid when another amino acid limits protein synthesis. A direct effect of protein synthesis on amino acid catabolism can be shown by a marked increase in amino acid catabolism when protein synthesis is inhibited. PMID- 8429384 TI - Species comparisons of tissue protein metabolism: effects of age and hormonal action. AB - Ontogenic responses in whole body and tissue protein metabolism are compared between ruminants and nonruminants. In sheep and cattle, the liver makes a smaller contribution to body protein mass and synthesis but is more responsive to age and nutrition than in the rodent. Protein synthesis increases within the gastrointestinal tract (to > 25% of total whole body protein synthesis) during the weaning phase in sheep and rats and in both species is sensitive to nutritional state. Protein synthesis in skin is also sensitive to intake but the sheep has a slower ontogenic decline compared with rats. Translational efficiency (g protein synthesis.d-1.g RNA-1) is unchanged for ovine muscle during the transition from the milk-fed to weaned state but is lower for the suckling rat. In contrast, in growing ruminants muscle translational efficiency decreases, whereas for the rat it is maintained. In young rodents, muscle protein synthesis is sensitive to nutrition and insulin, but the latter has little effect in sheep or indeed in nongrowing adult rats and humans. In ruminants the chronic response in muscle protein synthesis to intake may be exerted through the growth hormone and insulin-like growth factor 1 axis, although conclusive proof is lacking. PMID- 8429385 TI - Overview of retinoid metabolism. AB - Substantial and exciting progress has been made in understanding the cytoplasmic metabolism and the nuclear functions of vitamin A. Keys to this understanding have come from studies of the plasma and cytoplasmic proteins that bind retinol and retinoic acid, thereby regulating retinoid concentration and specifying the enzymatic reactions through which retinol is absorbed, stored as esterified retinol or oxidized to retinoic acid. In the nucleus, the hormone-like activity of retinoic acid is transduced by a family of retinoic acid receptors that regulate a large number of retinoic acid-responsive genes. The coordination of cytoplasmic retinoid metabolism and the nuclear effects of retinoic acid or other retinoid metabolites is now recognized as critical to embryonic development and the control of cellular differentiation throughout life. PMID- 8429386 TI - Retinoid metabolism during intestinal absorption. AB - A number of enzymes have been described that might participate in the metabolism of dietary carotenes and retinyl esters necessary for absorption. For several steps two or more different activities are known. Which of the candidate enzymes actually is important in normal physiology is generally not known with any certainty. However, various properties of the enzymes, determined in vitro, can suggest the more likely participants. The ability to interact with retinoids bound to the enterocyte-specific cellular retinol-binding protein, type two, is an important determinant in several cases. PMID- 8429387 TI - A comparison of the uptake, metabolism and biologic effects of retinol delivered to human keratinocytes either free or bound to serum retinol-binding protein. AB - In response to body demands, retinol (ROL) is secreted from the liver into the circulation bound to serum retinol-binding protein (RBP). The mechanism by which ROL is transferred from RBP to target cells remains controversial. To study ROL delivery, we have used a model system of cultured human foreskin keratinocytes (HKc) to compare the uptake, metabolism and biologic effects of ROL added either directly to the medium or bound to RBP. [3H]ROL added directly to the medium was rapidly taken up by HKc, and maximal accumulation of [3H]ROL occurred by 3 h. In contrast, [3H]ROL delivered bound to RBP was taken up very slowly but at a linear rate for at least 72 h. Several experimental approaches indicated that ROL uptake from RBP was not mediated by a cell surface receptor for RBP. The rate and extent of [3H]ROL metabolism to retinyl esters was the same whether the ROL was added directly to the medium or bound to RBP. In addition, several biologic responses by HKc to ROL showed the same dose response curves whether the ROL was added directly to the medium or bound to RBP. Overall, the results support a model of ROL uptake from RBP in which the vitamin is first slowly released from RBP into the aqueous phase and then becomes cell associated. In this manner, the cells are provided with a slow but constant supply of ROL and are protected from the rapid and potentially toxic accumulation of ROL that occurs in the absence of RBP. PMID- 8429388 TI - Overview of pup in a cup model: hepatic lipogenesis in rats artificially reared on a high-carbohydrate formula. AB - The artificial rearing technique allows nutritional investigations to be conducted in rat pups during a critical period that previously has been inaccessible to researchers. The technique will be useful for identifying dietary components contributing to metabolic adaptations in the preweaning period as well as "metabolic imprinting" or permanent metabolic effects in adult rats resulting from early dietary intervention. Artificially reared rat pups fed a formula high in carbohydrate-derived energy in the preweaning period have the following characteristics compared with pups fed a high-fat formula or reared naturally: (i) a higher level of plasma insulin, (ii) an increased hepatic lipogenic capacity and (iii) precocious induction of hepatic malic enzyme. The results also show that early exposure to a high-carbohydrate diet in the preweaning period predisposes the rat to an increased lipogenic capacity in liver and adipose tissues and to the development of obesity later in adult life. PMID- 8429389 TI - Use of pup in a cup model to study gastrointestinal development: interaction of nutrition and pituitary hormones. AB - The novel technique of artificial rearing (AR) of rat pups circumvents the difficulty of controlling diet composition and caloric intake. For studies of effects of nutrition and hormone interactions on gastrointestinal development, an appropriate experimental approach is to use AR rats whose corticosterone production is inhibited or abolished. Hypophysectomized (Hx) rats were used to examine whether growth retardation after Hx results from reduced caloric intake. Hx, sham-Hx and intact rats were isocalorically fed a cow-milk formula from day 12 to 18. Mother-fed (MF) Hx and intact rats were used as baseline controls. MF Hx showed retarded intestinal growth, decreased body weight gain and reduced skeletal growth. In contrast, AR-Hx rats showed intestinal hypertrophy, normal body weight gain and reduced skeletal growth. Intestinal lactase activity remained higher in MF-Hx or AR-Hx rats than in control groups. AR-Hx rats showed no precocious increase of intestinal maltase and sucrase activity as did AR controls. Trace levels of serum growth hormone was detectable in MF-Hx but not in AR-Hx rats. We conclude that caloric intake can promote intestinal and somatic growth in the absence of the pituitary gland and pituitary hormones are required for skeletal growth and intestinal enzymic differentiation. PMID- 8429390 TI - Use of pup in a cup model to study brain development. AB - The pup in a cup artificial rearing procedure allows for experimental manipulation of brain development during the early postnatal brain growth spurt in the rat that coincides with that of the human third trimester. The technique has been used to demonstrate temporal windows of vulnerability to alcohol; the brain appears to be vulnerable to alcohol after most of the neuronal populations have been generated and are starting to differentiate. Alcohol exposure via gastrostomy feeding tubes during the third trimester equivalent results in alterations in neuronal circuitry and deficits in neuronal populations. Some of the alterations are permanent. Importantly, studies incorporating artificial rearing of rat pups have demonstrated that the pattern of alcohol exposure is a critical factor for inducing alcohol-related brain damage; brain damage is increased with peak blood alcohol concentrations. PMID- 8429391 TI - The colostrum-deprived piglet as a model for study of infant lipid nutrition. AB - The large proportions of arachidonic acid (20:4 omega-6) and docosahexaenoic acid (22:6 omega-3) in brain and retina structural lipids are important for normal central nervous system function. Study of dietary requirements for omega-6 and omega-3 fatty acids for brain growth is difficult in the human infant because of variabilities in human milk fatty acids, lack of correspondence between blood lipids and central nervous system (CNS) lipids and ethical limits of tissue analyses. Comparison of nutrient requirements, lipid digestion, absorption and transport and milk and brain lipids and the timing of intestinal maturation and the brain growth spurt during fetal-infant development show the piglet to be an excellent choice for studies relevant to lipid nutrition of term gestation infants. Of practical relevance, piglets are easily hand-fed and, with a relatively large body size and rapid growth, provide ample tissue for analyses of the effects of diet fat on specific CNS cell membranes. Use of the piglet to define omega-6 and omega-3 fatty acid requirements and the effects of addition of marine oils to formulas and the lactating mother's diet on the developing CNS are described. PMID- 8429392 TI - Use of the colostrum-deprived piglet to evaluate parenteral feeding formulas. AB - Several different neonatal and infant piglet models have been invaluable as animal models in nutrition research. A preterm colostrum-deprived piglet model has been developed that is delivered at the desired gestational age, is cared for using the standard of care provided preterm human neonates including procedures for nutritional support and can be studied during pathological conditions induced under controlled conditions. The absolute values for each nutrient requirement would not be expected to be identical for preterm piglets and preterm human neonates. However, the effect of different levels of gestational maturity and superimposed pathophysiologies on nutrient requirements should be similar in the piglet and human neonate. PMID- 8429393 TI - The piglet can be used to study the effects of parenteral and enteral nutrition on body composition. AB - Measurement of body composition provides insight into the regulation of normal development as well as the interactions among nutritional, physiological, and biochemical functions. Studies of body composition require an animal model in which body composition at birth and postnatal changes are comparable to those in human infants. Essential to studies of body composition is an animal model that can be fed under various conditions (parenteral and enteral) and can undergo various measurements (whole body and cellular level). The piglet meets these requirements. The postnatal changes in the body composition of infants and piglets are reviewed, and various studies are discussed to show the effect of parenteral and enteral nutrition on piglet body composition. Confirmation of physiological and biochemical similarities in the regulation of body composition in human infants and piglets awaits further study. PMID- 8429394 TI - A review of study designs and methods of dietary assessment in nutritional epidemiology of chronic disease. AB - Increasingly the role of diet in the etiology of chronic disease has been the focus of research. Reviewed here are the epidemiologic study designs used for such research and the methodologies used for measurement of diet. Strengths and weaknesses of ecologic, cross-sectional, case-control cohort studies and clinical trials are discussed. Additionally the advantages and disadvantages of dietary recalls, food records, diet histories and food frequency questionnaires are reviewed as pertains to their use in nutritional epidemiology research. PMID- 8429395 TI - Issues in the long-term evaluation of diet in longitudinal studies. AB - Longitudinal studies are very useful for studying diet/disease relationships. The fundamental components of a longitudinal study are that: 1) data are collected for two or more distinct time periods; 2) the subjects are the same or comparable from one time period to the next; and 3) data are compared between or among time periods in the analysis. A longitudinal study is often assumed to be synonymous with a cohort study, but there are at least four possible definitions for a longitudinal study. While focusing on cohort studies, the paper describes the nature of longitudinal studies, including a discussion of how the different definitions differ from a cohort study and a set of important assumptions necessary to cohort studies. It also highlights some of the major issues associated with such studies, including the selection of a dietary survey methodology; data collection issues in multicultural, multilingual societies; the importance of nutrient databases; measurement error and misclassification in nutrient intake and energy adjustment. PMID- 8429396 TI - Epidemiology of calcium and vitamin D in bone loss. AB - Questions remain about the role of diet and skeletal bone loss. There is some indication from studies in premenopausal women that increased dietary calcium intake may be influential in promoting greater peak bone mass, although the evidence is inconsistent. A number of these studies have been reviewed recently. The findings from the clinical trials suggest that calcium supplementation as an effective intervention agent for bone loss in postmenopausal women must be more carefully investigated to identify at-risk women who will benefit from supplementation. The baseline levels of calcium intake and years since menopause should be considered. There is potential for a mild-to-moderate vitamin D deficiency leading to occult osteomalacia that contribute significantly to the excess bone loss and fracture observed in Caucasian women: The effects of diet, specific nutrients and metabolic bone disease may be difficult to ascertain when either the nutrient exposure is not well characterized or when the disease outcome is not well characterized. Additional work is needed to describe the relative influence of nutrient intake at different stages of the life cycle, the interactive nature between nutrients as they are expressed in bone metabolism and the role of nutrients according to level of hormones and exercise. PMID- 8429398 TI - Nutritional epidemiology of cervical neoplasia. AB - There appears to be a natural progression of some preinvasive cervical lesions to more advanced lesions. Although research has evaluated the associations between nutrients and particular preinvasive lesions or invasive disease, little work has been done to compare results across the spectrum of lesions in the progression to invasive disease. This review compiles studies that have evaluated the relation between nutrients and cervical neoplasia and evaluates the general consistency of the literature across stage of disease. Preformed vitamin A does not appear to be related to risk of any preinvasive or invasive lesions, whereas vitamin C has been associated with a reduced risk for dysplasia, in situ cancer, and invasive disease, particularly among smokers. There was evidence of reduced risks associated with various carotenoids and vitamin E at all stages of the disease process, although these studies were inconsistent and further work is needed. Folate was the only nutrient that appeared to be protective for dysplastic lesions and not related to risk of in situ or invasive disease. Red blood cell folate was a better predictor of dysplasia than were serum or dietary folate, and further investigation using this marker of folate status is warranted. Research is needed across a spectrum of lesions within one study, with particular attention to interactions between nutrients and other risk factors for disease. PMID- 8429397 TI - Colon cancer--do the nutritional epidemiology, the gut physiology and the molecular biology tell the same story? AB - Colon carcinogenesis models exist at epidemiologic, physiologic and molecular biologic levels. Thinking about the coherence of such models is useful both to inform colon cancer research and because the reasoning process may be generalizable. The consistent epidemiologic risk factors are low vegetable/fiber and high fat/meat/protein intakes. Others include physical activity, alcohol and reproduction. These epidemiologic risk factors appear to map to physiologic variables that provide mechanistic explanations for the associations: higher bile acids, fiber fermentation and effects of specific anticarcinogens found in vegetables. The possibility that the meat/fat association is due to carcinogens or promoters produced in cooked foods adds complexity to the physiologic model. As a link across genetics, physiology and epidemiology, the role of acetylator status is considered. Finally, whether relationships might exist between the epidemiologic/physiologic risk factors and the recently described molecular genetic changes and other colon cancer molecular mechanisms is considered. PMID- 8429399 TI - Body composition and aging: a study by in vivo neutron activation analysis. AB - Human body composition can be organized into five levels; atomic, molecular, cellular, tissue-system and whole body. Six elements (carbon, nitrogen, calcium, potassium, sodium and chlorine) can be directly measured in vivo at the atomic level using three neutron-activation systems at Brookhaven National Laboratory. When combined with an estimate of total body water, the six elements can be used to quantify the major chemical components at molecular level. In the present report, we first describe the neutron-activation approach to evaluating chemical compartments in vivo. Then, we present an example of how in vivo estimates of human chemical composition can be used to study the validity of two-compartment indirect methods of quantifying total body fat in the elderly. Our studies and the work of other investigators at Brookhaven National Laboratory suggest that neutron activation analysis provides an important opportunity to study the relation between aging and changes in elemental and chemical composition of humans in vivo. PMID- 8429400 TI - Soft tissue composition and bone mineral status: evaluation by dual-energy X-ray absorptiometry. AB - Dual-energy X-ray absorptiometry (DXA) is a new method for assessing whole-body and regional bone, fat and fat-free mineral-free soft tissue masses. X-rays at two discrete energy levels are collimated and directed into the body. The attenuation of the X-rays by the various chemical components in the body permits determination of important compositional variables. The precision and accuracy of the DXA measurements of bone mineral content and density are 99% and < 1% error, respectively. The reproducibility of the determinations of soft tissue composition is approximately 99%. Because the radiation dose associated with the X-ray exposure is low, DXA is a safe method for routine use in humans. Various applications are described, including an assessment of bone mineral status and body composition in adults and infants. This method is appealing for nutritional studies because it directly determines the composition of bone and of the body. PMID- 8429401 TI - Imaging techniques and anatomical body composition in aging. PMID- 8429403 TI - Aging and body composition: possibilities for future studies. AB - Three approaches are being taken to the development of techniques for measuring body composition: 1) improving present methods, 2) formulating and devising new body composition measuring techniques and 3) focusing on specific problems of body composition changes in aging. The improvement of multi-frequency bioelectrical impedance, computerized tomography, magnetic resonance and dual photon absorptiometry imaging belong under the first category. Recent developments in low radiation exposure measurement of body carbon and oxygen by neutron inelastic scattering and body protein by gamma-ray resonance are examples of new technology applied to the direct assessment of body composition of the elderly. The third category remains the most important one and should motivate the evolution of existing methods and the development of new. Current questions focus on distribution of regional body fat and its role in cardiovascular diseases and the depletion of lean body mass with age and its relation to functional capacity, quality of life and resistance to injury or disease. The relationship between the distribution of fat, the depletion of muscle tissue and changes in cellular function with age will become the focus of the future studies of body composition. PMID- 8429402 TI - Bioelectric and anthropometric assessments and reference data in the elderly. AB - Most studies using bioelectric impedance have estimated the volume of total body water, fat-free mass and on occasion extracellular fluid volume using whole-body resistance and reactance. However, the validity of bioelectric impedance has not been established for the elderly except in small samples of limited age ranges. The estimation of whole-body composition from impedance of body segments may be an alternative method that can be applied to many chair- and bedfast elderly patients. Also, measures of bioelectric impedance at lower and higher frequencies than 50 kHz are reported to differentiate the proportions of intra- and extracellular fluid volumes, and this could also provide important information about fluid quantities and balances in the elderly. Distribution statistics for body measurements for persons 65-85 y of age are available from the national health surveys and large clinical studies but reference data for persons > or = 80 y are sparse. Some reference data may be from groups that are not representative of the elderly because the samples were from cohorts older than elderly persons living today and significant group and racial differences existed among these as children and adults. These differences may have increased interindividual variances among the elderly. Suitable reference data for the present generations of elderly in the United States will be available with the culmination of NHANES III after 1994, but limited reference data for elderly persons up to 90 y of age have been published.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429404 TI - In vivo animal models of body composition in aging. AB - We developed several techniques that provide data on body elemental composition from in vivo measurements in rats. These methods include total body potassium by whole-body counting of endogenous 40K; total body calcium (TBCa), sodium and chloride by in vivo neutron activation analysis and total body phosphorus (TBP) and nitrogen (TBN) by photon activation analysis. These elements provide information on total body fat, total body protein and skeletal mass. Measurements were made in 6-, 12- and 24-month-old rats. TBN increased slightly between 6 and 12 months but was significantly lower by 24 months, indicating a substantial loss in total body protein. Working at the National Synchrotron Light Source, we studied rat femurs by computed microtomography (CMT), and the elemental profile of the femur cortex by synchrotron-radiation induced X-ray emission (SRIXE). Although there were no significant changes in TBCa and TBP, indices of skeletal mass, CMT revealed a marked increase in the size and number of cavities in the endosteal region of the femur cortex with increasing age. The SRIXE analysis of this cortical bone revealed a parallel decrease in the endosteal Ca/P ratio. Thus, there are major alterations in bone morphology and regional elemental composition despite only modest changes in total skeletal mass. PMID- 8429405 TI - Sarcopenia and age-related changes in body composition and functional capacity. AB - Advancing adult age is associated with profound changes in body composition. One of the most prominent of these changes is sarcopenia, defined as the age-related loss in skeletal muscle mass, which results in decreased strength and aerobic capacity and thus functional capacity. Sarcopenia is also closely linked to age related losses in bone mineral, basal metabolic rate and increased body fat content. Through physical exercise and training, especially resistance training, it may be possible to prevent sarcopenia and the remarkable array of associated abnormalities, such as type II diabetes, coronary artery disease, hypertension, osteoporosis and obesity. Using an exercise program of sufficient frequency, intensity and duration, it is quite possible to increase muscle strength and endurance at any age. There is no pharmacological intervention that holds a greater promise of improving health and promoting independence in the elderly than does exercise. PMID- 8429406 TI - Hormones, cytokines and body composition: can lessons from illness be applied to aging? AB - Loss of lean body mass (LBM) is a hallmark of aging and of acute and chronic illness. Loss of more than 40% of lean mass is not compatible with life. The causes of loss of LBM in aging remain obscure, although changes in growth hormone production, physical activity and the cytokines interleukin-1 beta and tumor necrosis factor-alpha may play a role. The usefulness of a disease model based on rheumatoid arthritis (RA) is considered, in which all these changes occur to a greater degree and independently of age. RA is a common autoimmune condition in which cytokine production is increased, LBM is reduced and mortality is greatly accelerated. These observations suggest that, with respect to body composition at least, RA represents a model of "accelerated" aging. A hypothesis is presented that unifies metabolic and immunologic observations and changes in body composition. The interrelationship of the immune system and body composition is an important area that deserves further investigation. PMID- 8429407 TI - Energy expenditure, aging and body composition. AB - We investigated the importance of adaptive variations in energy expenditure in the regulation of energy balance in young and elderly men of normal body weight leading unrestricted lives. Changes were measured in total and resting energy expenditure and body composition in response to overeating by approximately 4.2 MJ/d or undereating by 3.3 MJ/d for 21 d in 39 young and elderly men consuming a typical diet. In the young men, an average of 85-90% of the excess energy intake during overeating was deposited. During underfeeding, mobilization of body energy compensated for an average of 65% of the energy deficit in these subjects. The resting metabolic rate, averaged for fasting and fed states, increased during overfeeding (+0.63 +/- 0.20 MJ/d, P < 0.01) and decreased during underfeeding ( 0.42 +/- 0.12 MJ/d), but at least some of these changes were due to the obligatory expenditure associated with nutrient assimilation. There was no significant change in energy expenditure for physical activity or thermoregulation from overfeeding or underfeeding. Preliminary results indicate that aging had no effect on responses to overfeeding or underfeeding. These findings suggest that adaptive variations in energy expenditure are not the major determinant of energy regulation in young adults of normal body weight and that, instead, energy balance is maintained on a day-to-day basis through the control of nutrient intakes. PMID- 8429408 TI - Breastfeeding quadruplets. PMID- 8429409 TI - Preventing neonatal kidnapping. PMID- 8429410 TI - Understanding obesity in women. AB - Obesity is a complex health problem and, because of the incidence of obesity in women, a common concern for providers of women's health care. For many women, obesity becomes an obsession that leads to physiologic, psychologic, and social problems. To address the needs of the obese client better, nurses must acquire a comprehensive, research-based understanding of obesity. To develop a nursing care plan, the nurse must assess the client's diet history, level of motivation to effect a change in life-style, self-image, and self-esteem. PMID- 8429411 TI - Nurse-managed postpartum home care. AB - In 1989, Professional Nurse Associates, Inc., and Kaiser Permanente of Ohio collaborated to provide a home-centered postpartum recovery program to meet the postdelivery health-care needs of mothers, neonates, and families after a shortened hospital stay. This article reviews the history, process, and outcomes of that joint effort. The authors describe outcomes in terms of type and frequency of nursing diagnoses found on home visits, readmission rates, cost savings, and consumer satisfaction. PMID- 8429412 TI - Pregnant inmates: risk factors and pregnancy outcomes. AB - OBJECTIVE: To document the risk factors and outcomes of pregnant women incarcerated in a maximum-security prison. DESIGN: Descriptive correlational study. SETTING: Women's correctional facility. PARTICIPANTS: Eighty-nine pregnant women incarcerated during the third trimester of pregnancy. MAIN OUTCOME MEASURES: Medical, obstetric, life-style, and psychologic risk factors during pregnancy; birth weight; weeks of gestation at delivery; and presence of congenital anomalies in the newborn. RESULTS: Numerous risk factors during pregnancy, including chemical dependency, poor nutritional status, poor obstetric histories, high levels of anxiety and depression, and inadequate prenatal care. CONCLUSIONS: There is a need for intensive prenatal education for incarcerated women and for chemical dependency treatment programs designed specifically for incarcerated pregnant women. Interventions that address psychologic distress also are needed. PMID- 8429413 TI - Nurses' judgments of pain in term and preterm newborns. AB - OBJECTIVE: To examine neonatal intensive-care unit nurses' judgments of pain intensity in term and preterm newborns and to describe the cues used to assess the possible presence of pain in newborns. DESIGN: A combination of quantitative and qualitative approaches that included both parametric and nonparametric statistical procedures and a content analysis. SETTING: A large, university affiliated, tertiary-care hospital in central Canada. PARTICIPANTS: Forty-five registered nurses employed in the neonatal intensive-care unit. INTERVENTIONS: Nurses viewed videotapes of term and preterm newborns and written vignettes. Visual analogue scales and open-ended questions were used. MAIN OUTCOME MEASURES: Pain intensity ratings of full-term and preterm newborns and the cues identified as indicative of pain in newborns. RESULTS: Nurses gave significantly higher pain intensity ratings to the full-term group than to the preterm group of newborns. Some differences were found in the cues identified by the nurses in assessing pain in term versus preterm newborns. CONCLUSIONS: Nurses' judgments of pain intensity in newborns are influenced by the vigor and richness of the newborn's behavior response. Neonatal nurses must be provided with continuing education that focuses upon recognition of the signs and symptoms of pain in newborns. PMID- 8429414 TI - Family coping and premenstrual symptomatology. AB - OBJECTIVE: To examine how families cope when the woman experiences premenstrual symptomatology. DESIGN: A descriptive panel design and purposive, nonprobability sampling through interviews and questionnaires. SETTING: Participants' homes. PARTICIPANTS: One hundred four couples who met the study criteria were recruited from medical practices and community seminars and through media announcements. MEASURES: Study outcome measures were formulated after data collection. RESULTS: High-symptom families used spiritual coping strategies more frequently than low symptom families (R = .427, p = 0.003 in women, p < 0.001 in men). Husbands were more likely than wives to report that their families coped by believing that the problem would go away if they waited long enough (t = -3.06, p = 0.003). Husbands of women with high symptomatology reported that their families used this passive approach to a significant degree (t = -3.43, p = 0.001). CONCLUSIONS: Religious or spiritual support may be especially important to families in which the woman suffers from premenstrual symptomatology. These families may also keep problems within the family and be socially detached. Nurses may help by encouraging the use of social supports, religious or spiritual support, and other family resources. PMID- 8429415 TI - Validation of the nursing diagnosis of ineffective breastfeeding. AB - OBJECTIVE: To test the validity of the definition, related factors, and defining characteristics of the nursing diagnosis of ineffective breastfeeding. Also, to test whether defining characteristics relevant to the interruption of breastfeeding may be included in the diagnosis of ineffective breastfeeding. DESIGN: A methodologic study using a two-stage Delphi process. PARTICIPANTS: Sixty-six nurses in the first round of the Delphi process and 34 nurses in the second round. MAIN OUTCOME MEASURES: A Likert-like scale was used to measure nurse experts' opinions as to how reflective each defining characteristic was of ineffective breastfeeding. RESULTS: The definition of ineffective breastfeeding was simplified, related factors were added, and defining characteristics were clarified. Two characteristics of interruption of breastfeeding were found to be relevant to ineffective breastfeeding, but were considered minor rather than major. CONCLUSION: The definition, related factors, and defining characteristics of ineffective breastfeeding have been clarified and validated by nurse experts, but they need further validation by breastfeeding women. PMID- 8429416 TI - Twin transfusion syndrome. AB - Twin transfusion syndrome (TTS) is a rare, yet frequently fatal, complication of twin gestations. Recently published data demonstrate a reduction in the perinatal mortality rate associated with TTS from nearly 100% to 20-40%. This article explains the pathophysiology of TTS, newly described treatments, complications, and specific aspects of nursing care for patients with this complication of multiple gestation. PMID- 8429417 TI - Idiopathic cervical lesions: in vivo investigation by oral microendoscopy and scanning electron microscopy. A pilot study. AB - The development and progression of cervical lesions have not been thoroughly documented. Clinically, two morphologically distinct forms, saucer-or wedge shaped, have been described. In this study, the micromorphology of manifest cervical lesions was documented in six subjects, by SEM replication and microendoscopy. Optoelectronic recordings were made of extreme mandibular lateral excursions. Salivary parameters were evaluated and a detailed case history, including oral hygiene and dietary habits, was taken. Lesions of varying severity could be observed in the same subject. Longitudinal fractures of the enamel were common. The dietary analyses were uneventful and salivary values were normal. No correlation was found between brushing habits and the location and severity of the lesions. The optoelectronic recordings, however, indicated a possible correlation between irregular lateral excursion and the severity and location of the lesions, with a tendency for fewer and less severe lesions on the preferred chewing side. The results have indicated some areas of interest for future studies applying non-invasive observation methods. PMID- 8429418 TI - The ratio between vertical and horizontal changes of impressions. AB - The purpose of this study was to analyze theoretically and experimentally the relations between the horizontal and the vertical changes of impressions taken with a mandibular stock tray. According to the theoretical analysis the ratio between the horizontal (delta X) and the vertical changes (delta Z) can be expressed as: [formula: see text] where mu is the Poisson's ratio and Lx and Lz are the horizontal and vertical dimensions of the die, respectively. With regard to the experimental findings the Poisson's ratio calculated by this expression was mu = 0.44 which fits the range of Poisson's ratio for rubber materials. In addition it was shown that the ratio between the vertical and horizontal strains is constant for a particular impression technique. PMID- 8429419 TI - The effects of dentine bonding agents on marginal leakage of composite restorations--long-term studies. AB - The aim of this study was to compare the effects upon marginal leakage of a number of dentine bonding agents when used with a posterior composite resin, Occlusin, after storage for periods of up to 2 years. The bonding agents used were Gluma, Scotchbond, Topaz and an experimental material. The results were compared with a glass ionomer, Chemfil II. Class V cavities with or without a bevelled cavo-surface margin were prepared in the buccal surfaces of extracted premolar teeth. Following restoration, the teeth were stored for periods of up to 2 years and then thermally cycled. Marginal leakage was subsequently demonstrated using a radioactive isotope containing 45Ca, and an auto-radiographic technique. Image analysis was used to determine the total amount of linear leakage for each specimen. The results showed that some leakage occurred for all materials at each time interval. The bevelled design of cavity allowed significantly less leakage than the unbevelled cavity. Overall, there was little difference between the various dentine bonding agents. The glass ionomer restorations showed significantly less leakage than the composite resin and dentine bonding agent combinations in unbevelled cavities. PMID- 8429420 TI - Posterior palatal seal adaptation: influence of processing technique, palate shape and immersion. AB - Accuracy of fit of denture bases is critical to adequate retention. This study compared the dimensional change of a newer continuous-injection technique with a standard trial-pack technique as determined by measuring the posterior palatal border opening. The influence of palate shape and immersion were also assessed. Stone casts were made from master moulds with either a high or flat palate. Denture base adaptation was measured at 5 mm intervals across the entire posterior palatal border. Measurements were made after deflasking, trimming and polishing, and after immersion in room temperature water for 1 h, 1 day and 1 week. Statistical analysis (P < 0.05) showed that where differences occurred between the two techniques, openings were always smaller for the continuous injection technique. It was also shown that dimensional changes were always larger for the flat palate compared with the high palate. Dimensional change using the continuous-injection technique was reduced by immersion in water, while no influence was observed for the trial-pack technique. It was concluded that the continuous-injection technique showed smaller dimensional changes compared with the standard trial-pack technique, and that these changes were influenced by palate shape and immersion in water. PMID- 8429421 TI - The recording and analysis of EMG and jaw tracking. II. Reproducibility of jaw tracking. AB - In 10 male and 10 female adults jaw movements and muscle activity were monitored during chewing. Subjects had a Class I occlusion and fluent unrestricted jaw movements; non exhibited signs or symptoms of craniomandibular disorders. Recordings were made on two occasions separated by 2 weeks. Within each recording session the subjects carried out a number of tasks under direction, including eating nuts and gum. The jaw movements were monitored with a Sirognathograph which had been interfaced to a computer for data logging and analysis. A computer program identified each chewing stroke and after scrutiny a data base was constructed of measurements from a subset of 4447 suitable strokes. Mean values of 10 parameters selected to describe jaw movements during chewing were entered into an ANOVA. This indicated that the major contribution to variance was due to the subject from whom the data was recorded. This intersubject variability is well known, but can its separation from the other contributions to the variation within the data be assessed? To provide an estimate of the spread of data the variance for each parameter was calculated, transformed and these values entered into an analysis of variance. All subject differed significantly in their chewing movements from one another. But 15 of the 20 subjects showed a greater degree of consistency than the other five: they exhibited good inter- and intra-recording reproducibility. The food being eaten made a major contribution to the variability. The timing of the chewing strokes was also more consistent than the dimensions of the chewing envelope. PMID- 8429422 TI - The effect of time of trimming on the surface finish of anterior composite resins. AB - This in vitro study investigates whether the surface smoothness of a finished composite resin is a function of the time at which it is trimmed. Five materials were studied. Visiodispers (VIS) is a sintered microfine, the remainder were hybrid formulations: Miradapt (MIR), Command Ultrafine (COM) and two experimental formulations-'EXP (2)' and 'EXP (15)'-with larger filler particles. All except MIR were photo-activated. Samples were placed and cured in holes machined in Perspex rods, ensuring excess of material and moulded to shape with cellulose acetate matrix strip. The strips were removed immediately after light curing and after 5 min in the case of MIR. Sample groups were finished after the following intervals: 10, 15, 30, 45s, 1, 2, 3, 4, 5 min, dry, and after 1 and 7 days in 37 degrees C water. The finishing procedure used plain-cut tungsten-carbide burs and coarse, medium and fine finishing discs, which were applied for standardized times, directions and pressures. Surface roughness was assessed visually and with a surface profilling instrument. COM and VIS materials significantly improved in surface finish when the procedure was delayed for 7 days. However, EXP (2), EXP (15) and MIR surface finishes were independent of finishing time. This may result from a rapid attainment of optimum surface hardness, whereas with COM and VIS incomplete curing leads to smearing of resins and filler across the surface. PMID- 8429423 TI - The oral status of elderly removable partial denture wearers. AB - A random sample of 1019 elderly home dwelling persons participated in this study. There were 809 dentate respondents, 28.6% of whom wore removable partial dentures. They were examined for coronal and root caries, gingival recession, pocket depth and loss of gingival attachment. The removable partial dentures were also evaluated. Abutment teeth were found to be more likely to have caries and periodontal disease than all other teeth. Using a MANOVA statistical procedure, the analysis indicated that the partial denture itself, irrespective of any professionally determined problems with the denture, appears to affect coronal and root caries on the teeth of partial denture wearers. Partial dentures judged to need repair or adjustment were related to periodontal status. The data on adverse effects of partial dentures suggest a need for patient education by the dentist and through public health measures, and good oral self care and regular professional recall for people who wear removable partial dentures. PMID- 8429424 TI - Thermal diffusivity of glass ionomer cement systems. AB - The thermal diffusivity has been measured for 10 glass ionomer and resin-based materials: three conventional (water-hardened) glass ionomer cements, two silver reinforced glass ionomers, an experimental stainless steel-reinforced glass ionomer, three visible light-cured (VLC) glass ionomer-resin hybrid materials, and a VLC resin-based product developed for the same clinical uses as the hybrid materials. Cube-shaped specimens, c. 10 x 10 x 10 mm, initially at room temperature were immersed in mercury surrounded by an ice-water bath. From the experimental cooling curve a semi-log plot of relative temperature decrease vs. time yielded a straight line whose slope is proportional to the thermal diffusivity. The values ranged from 1.74-5.16 x 10(-3) cm2 s-1, and all of the materials tested would have adequate insulating properties provided normal clinical thickness levels for lining materials are maintained. It was found that the thermal diffusivities for the three metal-reinforced glass ionomers, where composition information is available, do not follow a rule of mixtures applied to the individual components. PMID- 8429425 TI - Longitudinal study on forces transmitted from denture base to retainers of lower distal-extension removable partial dentures with conus crown telescopic system. AB - The purpose of this study was to perform longitudinal measurement of forces transmitted from the denture base to retainers of lower distal-extension removable partial dentures with a conus crown telescopic system. The experimental denture was embedded with a force-detecting unit which could detect a change in lateral and vertical forces transmitted from denture base to retainers at the same time. Forces were measured during chewing of food on several separate occasions from the insertion of new dentures to about 3 months after. Max.ver (maximal mean value of vertical force) and Max.lat (maximal mean value of lateral force) were calculated from the peak level of all strokes of chewing of food at the minimum of the interocclusal distance. The results are summarized as follows: (i) There were no significant changes of Max.ver and Max.lat; (ii) There were no significant differences of Max.ver between subjects the value of which was about 20 N; (iii) There were significant differences of Max.lat between subjects the value of which was less than 15 N. PMID- 8429426 TI - The properties of metal-reinforced glass ionomer materials. AB - The physical properties and bond strengths of two glass ionomer materials reinforced with silver and amalgam alloy powders were compared with those of a conventional material from the same manufacture as well as two commercially available products. The diametral tensile strength, hardness and bonding strength are improved with the addition of either commercial available silver particles or fabricated high-copper amalgam alloy powders to the glass. Simple mixture of the metal or alloy powders with the glass ionomer cement seems to be feasible to improve the properties of the regular cement. However, further studies in formulating an optimal composition of metal or alloy, setting characteristics and long-term clinical evaluation are necessary before proposing uses for this new material. PMID- 8429427 TI - Impact fracture characteristics of intact and crowned human central incisors. AB - Dynamic fracture energies and patterns of fracture in extracted human central incisors were determined for groups of intact controls, groups with Vita Dur N crowns, Vita Hi Ceram crowns, Dicor crowns and porcelain veneers. Teeth were struck on their middle labial surfaces by a pendulum impact device. The mean fracture energy for teeth with Dicor crowns was significantly lower than for all other groups (P < 0.05). Control tooth crowns fractured obliquely in an apical direction. Vita Dur N and Dicor crowns shattered, the underlying tooth usually fracturing in the plane of the impact force. Vita Hi Ceram crowns chipped at the site of impact and some fractures were located in the roots. Gold crowns remained cemented and fracture occurred at the crown/root junction, or in the root. Porcelain veneers fractured at the site of impact but remained cemented. Dicor crowns were less fracture resistant than other restoration types tested. Porcelain veneers and full gold crowns stiffened teeth which led to more root fractures than the porcelain crowns. PMID- 8429428 TI - Effect of dental metals on magnetic resonance imaging (MRI). AB - The aim of this research was to examine the effects of metals used in dentistry upon magnetic resonance imaging (MRI). Ten disk-shaped samples of metals have been used in this study. Each sample has been set into a volunteer who had produced a normal MR image when tested. The subject was imaged in the sagittal, frontal and coronal plane with a 0.1 T MRI device and results examined for MR defects. Au, Ag, Au-Ag-Pd and amalgam produced no defect. Ni-Cr, Co-Cr and SUS304 expressed small amounts of MR defects, but SUS405, Pd-Co-Ni and Sm-Co expressed large defects. Those metals which create MR image degradation must not be used for fixed prosthesis or orthodontic devices both of which could easily be made from other metals. If the MR image degrading metals are to be used in dentistry, their application should be restricted to removable prosthesis. PMID- 8429429 TI - Molecular genetic advances in fragile X syndrome. AB - The fragile X syndrome is recognized as the most common heritable condition resulting in mental retardation. The disabilities are substantial, and therefore early detection is mandatory to assist with reproductive counseling of families in which the fragile X syndrome has occurred. Highly accurate, direct DNA diagnostic testing can now be performed to diagnose the fragile X syndrome without the involvement of individual family members, as was the situation with the use of DNA linkage analysis. Such testing is rapidly becoming a standard diagnostic tool for screening of individuals with suspected fragile X syndrome, of potential unaffected carriers, and of patients with undefined mental retardation. Fragile X testing should be considered for all children with developmental delay of unknown cause. Autistic children will occasionally be found to have mutations in FMR-1. Detection of affected individuals will allow early intervention for these individuals and will assist families with their reproductive decisions (including prevention) in subsequent offspring. An understanding of the molecular genetics of fragile X syndrome has resulted in the resolution of the Sherman paradox and is the first molecular characterization of a chromosomal fragile site, a finding that almost certainly will be important in understanding the cause of chromosomal rearrangements involving fragile sites. In addition, molecular details of the fragile X mutations have yielded insight into "heritable unstable elements," of which the fragile X chromosome is one of the first characterized examples. Thus a similar molecular mechanism involving a trinucleotide repeat may explain the genetics of myotonic dystrophy and spinal bulbar muscular atrophy (Kennedy disease); it seems reasonable to assume that other genetic diseases also may result from disruption of genes by inherited unstable elements. PMID- 8429430 TI - Human parvovirus B19-associated arthritis in children. AB - Human parvovirus B19 (HPV B19) infection has been associated with chronic joint complaints in adult patients. We now report 22 children with joint complaints associated with recent HPV B19 infection. These children had either erythema infectiosum or serologic evidence of recent infection. Twenty children had arthritis; two had arthralgias. Eleven children had associated constitutional symptoms. Laboratory findings were generally normal. The duration of joint symptoms was less than 4 months in 14 children; however, six children have had persistent arthritis for 2 to 13 months, which would fulfill criteria for the diagnosis of juvenile rheumatoid arthritis. Although HPV B19 is usually associated with acute arthritis of brief duration, in some children infection with HPV B19 may be associated with the development of chronic arthritis. PMID- 8429431 TI - Juvenile-onset mixed connective tissue disease: longitudinal follow-up. AB - To establish the symptoms and clinical course of juvenile-onset mixed connective tissue disease, we studied 14 patients, classified according the criteria of Kasukawa et al. The patient records were studied retrospectively and all patients were examined in a 1-day follow-up program. Systemic lupus erythematosus and polymyositis/dermatomyositis-like symptoms disappeared in time, whereas scleroderma-like symptoms (such as in the Raynaud phenomenon) and joint abnormalities persisted. Extensive limitation of joint function was found in four patients. At the time of follow-up, no active renal disease was found. Thrombocytopenia was still present in one of the three patients who had had this feature. All patients had abnormalities of esophageal motility. Long-term corticosteroid treatment was associated with aseptic bone necrosis in three patients and growth retardation in one. We conclude that the Kasukawa criteria are easy to apply to children, and that juvenile-onset mixed connective tissue disease has many similarities to the adult form of the disease. PMID- 8429432 TI - Interference of immune globulin with measles and rubella immunization. AB - Passively acquired antibody may interfere with the active antibody response to live viral vaccines such as measles and rubella. To evaluate the duration of this inhibitory effect, we measured the measles and rubella antibody responses of Apache children immunized with measles, mumps, and rubella vaccine at varying intervals after administration of an immune globulin termed bacterial polysaccharide immune globulin (BPIG). This specific immune globulin contained measles and rubella antibody titers similar to those in standard intramuscularly and intravenously administered immune globulins. Antibody responses to measles vaccine were inhibited for up to 5 months after a BPIG dose of 80 mg IgG per kilogram of body weight, but responses to rubella vaccine were inhibited for only 2 months. Most children who had a decreased measles antibody response to primary measles, mumps, and rubella immunization given 1 1/2 to 4 months after BPIG administration responded to a booster immunization given 6 months after their last BPIG dose. We conclude that high doses of immune globulin (> 10 mg/kg) may inhibit the antibody response to measles for more than 3 months. We propose that the interval between administration of immune globulin and measles and rubella immunization be adjusted on the basis of the dose of immune globulin. PMID- 8429433 TI - Antibody response to outer membrane protein of nontypeable Haemophilus influenzae in otitis-prone children. AB - One of the major outer membrane proteins of nontypeable Haemophilus influenzae, P6, is highly conserved among strains, serves as a target for bactericidal antibody, and has been proposed as a possible vaccine candidate. The serum antibody response to P6 was studied in otitis-prone and normal children by an enzyme-linked immunosorbent assay. Of 20 otitis-prone children, 12 (60%) had a serum IgG antibody response to P6 after otitis media; however, the mean acute antibody level for the group, 4.6 micrograms/ml, was not significantly different from the convalescent level, 5.4 micrograms/ml. Anti-P6 antibody levels were also measured longitudinally for 10 to 25 months in 30 otitis-prone and 13 healthy children. Antibody levels increased sevenfold in the normal group compared with less than three-fold for the otitis-prone group and were significantly higher in the normal children after the age of 18 months (p < 0.05). Finally, otitis-prone children who had two or more episodes of otitis media with nontypeable H. influenzae did not have an anamnestic antibody response to P6. The failure to recognize P6 as a specific immunogen may account for recurrent infections. Moreover, the data suggest that otitis-prone children may not respond adequately to a vaccine containing P6. PMID- 8429434 TI - Bone metabolism in children with asthma treated with inhaled beclomethasone dipropionate. AB - Previous studies have shown that inhaled corticosteroids can affect bone metabolism in adults. A study to assess the effect of inhaled beclomethasone, 300 to 800 micrograms/day for at least 6 months (mean 25 months), was therefore undertaken in children. In part 1 of the study, 18 children with asthma, aged 4 to 17 years (mean 10.1 years), were compared with an age- and sex-matched group of children with asthma not treated with corticosteroids. In part 2, eight more pairs were compared. Comparisons were also made with 61 healthy children. Bone mineral density measured by radiographic absorptiometry, and bone mineral content measured by single-photon absorptiometry and by dual-energy x-ray absorptiometry, showed no significant differences. Serum levels of calcium, magnesium, zinc, total alkaline phosphatase, bone specific alkaline phosphatase, parathyroid hormone, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D also showed no differences. The activity of tartrate-resistant acid phosphatase, a marker of bone resorption, was significantly lower in the beclomethasone group than in both the asthma control and the normal control groups, but urine calcium excretion did not differ. Patients with asthma had lower serum osteocalcin and higher serum copper levels than control subjects without asthma, but treatment with beclomethasone did not affect these values. We conclude that inhaled beclomethasone (up to 800 micrograms/day) does not reduce bone mineralization or increase bone resorption. Effects on bone formation were difficult to assess because asthma per se caused a significant reduction in osteocalcin, a sensitive marker of bone formation. PMID- 8429435 TI - Thyroxine-binding globulin deficiency detected by newborn screening. AB - We examined the results of the Northwest Regional Screening Program from May 1975 to June 1991 to determine the prevalence of inherited thyroxine-binding globulin (TBG) deficiency and its effect on thyroid hormone concentrations in infants. Serum thyroxine (T4), triiodothyronine resin uptake (T3RU), and thyrotropin values were requested of physicians caring for all infants with a single filter paper T4 level < 38.6 nmol/L (3 micrograms/dl) or a T4 level < 3rd percentile on two filter paper tests (at birth and 2 to 6 weeks of age). From 1,367,724 infants screened in five states, TBG deficiency, an X-linked disorder, was identified in 317 infants (285 boys). For the entire screening program the calculated frequency of TBG deficiency was 1:4315 infants (1:2400 for boys). In Oregon, where 95% of infants have two screening tests performed, the calculated frequency was somewhat higher (1:3080 infants; 1712 boys) and is probably more accurate. The mean serum T4 concentration for TBG-deficient boys was 41.9 nmol/L (3.26 micrograms/dl); 31% had values < 25.7 nmol/L (2.0 micrograms/dl). The mean serum T4 concentration for TBG-deficient girls was 60.2 nmol/L (4.68 micrograms/dl), with none < 2.0 micrograms/dl. The mean T3RU value was 0.472 in TBG-deficient boys, and 0.412 in TBG-deficient girls; the T3RU value was > 0.55 in 24% of TBG-deficient boys but was > 0.55 in only one girl. Free serum T4 levels were normal in all 56 TBG deficient infants studied, and TBG levels were low in all 20 infants studied. Inherited TBG deficiency is common in boys in the Northwest, with a frequency of 1:1700 and a male/female ratio of 8.9:1. Boys with TBG deficiency have mild, moderate, or severe alterations in total T4 and T3RU values, but severe deficiency is rare in girls. PMID- 8429436 TI - Limitations of an oscillometric ambulatory blood pressure monitor in physically active children. AB - OBJECTIVE: To determine the limitations of an oscillometric device for measuring ambulatory blood pressure in physically active children. DESIGN: Observational descriptive. SUBJECTS: Sixty-one children 4.3 to 18.7 years of age. INTERVENTIONS: Twenty-two high school students wore an ambulatory blood pressure monitor (SpaceLabs model 90202) for 24 hours. A subject-kept activity diary facilitated data interpretation. In 39 additional subjects, measurements were recorded during strenuous exercise, at rest, and after ambulation. Before and after the students wore the unit, oscillometric and auscultatory blood pressure measurements were simultaneously taken to calibrate the ambulatory blood pressure monitor, evaluate its accuracy, and assess deterioration in the calibration of the unit with use. RESULTS: During 24-hour ambulatory monitoring, 29% of the readings were edited because of error codes, primarily because of vibratory interference. At the end of the 24-hour monitoring period, simultaneous pressures by oscillometric and auscultatory techniques agreed within 6 mm Hg in 17 of 18 cases for both systolic and diastolic blood pressures. During exercise, 88.4% of the systolic and diastolic blood pressure readings agreed within 8 mm Hg. In 20 other ambulatory subjects with 80 simultaneous oscillometric and auscultatory measurements, all systolic and 88.5% of the diastolic values agreed within 10%. CONCLUSIONS: These results indicate that the SpaceLabs model 90202 unit provides a reasonably accurate assessment of ambulatory blood pressures in mildly active and inactive children and that blood pressure can be monitored with time. PMID- 8429437 TI - Growth in female gymnasts: should training decrease during puberty? PMID- 8429438 TI - Classification of growth hormone insensitivity syndrome. PMID- 8429439 TI - Energy expenditure in cystic fibrosis during activities of daily living. AB - Energy expenditure during various activities of daily living in normally nourished female adolescents with cystic fibrosis was compared with that in matched healthy control subjects. Energy expenditure at rest, during sitting and standing, and during two levels of exercise was increased significantly in patients with cystic fibrosis (122% +/- 14%) compared with control subjects (104% +/- 10%) (p < 0.05), but incremental increases from one level of activity to another did not differ. We conclude that the various activities of daily living are not responsible for increased energy needs in female adolescents with cystic fibrosis. PMID- 8429440 TI - Ovarian microcysts in girls with isolated premature thelarche. AB - We compared the frequency of ovarian microcysts in girls with premature thelarche (n = 27) to that in age-matched control subjects (n = 24). There was an increased prevalence of detectable ovarian microcysts in girls with premature thelarche. The presence or absence of cysts did not correlate with basal gonadotropin or estradiol levels. PMID- 8429441 TI - Calcium-deficiency rickets in a four-year-old boy with milk allergy. AB - A 4-year-old boy was found to have rickets associated with normal serum levels of 25-hydroxyvitamin D and high serum levels of 1,25-dihydroxyvitamin D. These findings were thought to be the result of dietary calcium deficiency caused by the prolonged elimination from his diet of cow milk and milk products because of allergy. Adequate intake of calcium resulted in rapid improvement. PMID- 8429442 TI - Acute mercury poisoning (acrodynia) mimicking pheochromocytoma in an adolescent. AB - A 14-year-old boy was seen because of irritability, insomnia, lethargy, and profuse sweating, together with hypertension (blood pressure: 160/120 mm Hg), tachycardia, and a diffuse erythematous rash with desquamation of the palms and soles. Initial biochemical investigation suggested a diagnosis of pheochromocytoma, but subsequently a history of exposure to mercury vapor was obtained. This case emphasizes the clinical and biochemical similarities between mercury poisoning (acrodynia) and pheochromocytoma. PMID- 8429443 TI - Diagnosis of occult muscular dystrophy: importance of the "chance" finding of elevated serum aminotransferase activities. AB - We report our experience with four children, including one girl, in whom the eventual diagnosis of muscular dystrophy was made because of persistent, unexplained elevated serum aminotransferase values. Measurement of serum creatine kinase activity and careful physical examination are the most useful and cost effective means of correctly identifying these patients. PMID- 8429444 TI - Estimates of uridine diphosphate hexoses in erythrocytes: implications for galactosemia. AB - Impurities in a reagent dehydrogenase caused overestimates of erythrocytic uridine diphosphate glucose and accounted for clinically important differences in results between those of one group of investigators using enzymatic methods and those of two other groups using enzymatic methods, high-performance liquid chromatography, and nuclear magnetic resonance. These data have relevance for the current debate regarding the pathophysiologic changes in galactosemia. PMID- 8429445 TI - Improved pulmonary outcome after exogenous surfactant therapy for respiratory failure in term infants requiring extracorporeal membrane oxygenation. AB - A blinded, randomized, controlled study was designed to test whether multiple dose surfactant therapy would improve pulmonary outcome in term infants with respiratory failure, resulting in a shortened period of extracorporeal membrane oxygenation (ECMO). Infants > or = 34 weeks of gestational age in severe respiratory failure and receiving ECMO were stratified by diagnosis and then randomly assigned to the treatment or the control group. Four doses of modified bovine lung surfactant extract (beractant) were administered to the surfactant group (n = 28), and an equal volume of air was administered to the control group (n = 28). Lung compliance was initially low in both groups; after treatment, values were higher with time in the surfactant group (F = 5.40, p = 0.026). The ECMO treatment period was significantly shorter in the surfactant group (mean +/- SD: 107 +/- 33 hours vs 139 +/- 54 hours for the control group; U = 232, p = 0.023). Tracheal aspirate concentrations of surfactant protein A were low in both groups, and then increased steadily to a higher level in the surfactant group (F = 2.58, p = 0.04). The overall incidence of complications after ECMO was decreased in the surfactant group (18% vs 46% for the control group; chi-square value = 5.004, p = 0.025). Radiographic scores, echocardiographic findings, incidence of intracranial or pulmonary hemorrhage and bronchopulmonary dysplasia, time to extubation, duration of oxygen therapy, and duration of hospitalization did not differ between the two groups. Beractant in this population improved pulmonary mechanics, increased surfactant protein A content in tracheal aspirate, decreased time on ECMO duration, and reduced disease complications. PMID- 8429446 TI - Reduced serum osteocalcin and 1,25-dihydroxyvitamin D concentrations and low bone mineral content in small for gestational age infants: evidence of decreased bone formation rates. AB - In small for gestational age (SGA) infants, bone mineral content (BMC) is low but the reasons are unclear and the possible relationships between calcium-regulating hormones and BMC have not been studied. We hypothesized that BMC would be lower and concentrations of serum parathyroid hormone and 1,25-dihydroxyvitamin D would be higher at birth in SGA infants than in appropriate for gestational age (AGA) infants. Forty-two term SGA infants and 126 term AGA infants, matched 1:3 specifically by gestation (+/- 1 week) and birth month, were studied prospectively. The BMC of the distal one third of the radius was measured before 3 days of age by photon absorptiometry. The BMC was lower in SGA than in AGA infants. Both SGA and AGA infants had lower BMC in summer or spring than in winter; BMC differences between groups remained significant after adjustment for season (p = 0.0001). Cord serum osteocalcin and 1,25-dihydroxyvitamin D values were lower in SGA than in AGA infants. There were no differences between groups in cord serum levels of intact parathyroid hormone, 25-hydroxyvitamin D, calcium, phosphorus, and magnesium. Relationships were positive between BMC and birth weight and were inverse between BMC and intact parathyroid hormone values. We suggest that reduced uteroplacental blood flow in SGA infants may result in reduced fetal-placental production of 1,25-dihydroxyvitamin D, which results in low BMC and low serum osteocalcin values; fetal serum parathyroid hormone values may be relatively elevated because of reduced placental mineral supply. PMID- 8429447 TI - Controlled trial of beclomethasone dipropionate by nebulization in oxygen- and ventilator-dependent infants. AB - Parenteral glucocorticoids have been shown to be effective in the treatment of oxygen- and ventilator-dependent bronchopulmonary dysplasia. We conducted a randomized, prospective study using a nebulized, water-soluble form of beclomethasone dipropionate for the treatment of infants with oxygen- and ventilator-dependent lung disease. Newborn infants with chest x-ray changes consistent with bronchopulmonary dysplasia at 14 days of age were randomly assigned, in a paired sequential fashion by birth weight, to treatment (beclomethasone) or placebo (saline solution) groups. Treatment included three nebulized doses of beclomethasone (50 micrograms) or saline solution per day for 28 days. Measured variables included tidal volume, total dynamic compliance, and airway resistance. Weight gain, gender, and incidence of infection during therapy were also recorded. Pulmonary functions were measured before initiation of therapy and weekly thereafter. Thirteen infants, seven in the saline solution group and six in the beclomethasone group, met study criteria and completed treatment. Infants treated with beclomethasone had reductions in airway resistance that were significant in weeks 2, 3, and 4 (p < 0.05, p < 0.02, and p < 0.001, respectively). Dynamic lung compliance increased at weeks 3 and 4 (p < 0.01 and p < 0.05, respectively). As expected, tidal volume increased with weight and time, but there were no significant differences between groups. There were no differences between the groups in weight gain, gender, or infection. This study demonstrates that beclomethasone by nebulization (1) reduced airway resistance in oxygen-dependent neonates with bronchopulmonary dysplasia, (2) improved dynamic lung compliance, as reported with parenterally administered glucocorticoids, and (3) produced no apparent increase in the incidence of infection. PMID- 8429448 TI - T-cell-depleted maternal bone marrow transplantation for siblings with X-linked severe combined immunodeficiency. AB - Prenatal diagnosis of X-linked severe combined immunodeficiency in a male fetus made possible elective neonatal bone marrow transplantation before onset of symptoms. Bone marrow transplantation was performed by using T-cell-depleted maternal marrow that had been cryopreserved 2 years earlier, at the time that his older affected brother underwent transplantation. The second patient had less morbidity and more rapid reconstitution of his immune function. PMID- 8429449 TI - Treatment of severe familial hypercholesterolemia in childhood with sitosterol and sitostanol. AB - This study was undertaken to compare the ability of two plant sterols to reduce serum levels of lipids and to compare their mechanism of action in nine children with severe familial hypercholesterolemia (total and low-density lipoprotein cholesterol concentrations averaged 9.57 mmol/L (370 mg/dl) and 7.87 mmol/L (301 mg/dl)). After a 3-month strict diet, the children were given sitosterol pastils (2 gm three times a day) for 3 months, followed by a 7-month course of sitostanol (0.5 gm three times a day). Serum lipoprotein levels and serum concentrations of campesterol and sitosterol were determined in all nine children, and the fecal excretion of neutral and acidic sterols were determined in seven children at the end of each therapeutic regimen. Sitosterol reduced low-density lipoprotein cholesterol levels by 20% (p < 0.01); sitostanol reduced low-density lipoprotein cholesterol levels by 33% after 3 months and 29% after 7 months (p < 0.01 compared with diet; p < 0.05 compared with sitosterol). Although sitosterol did not alter serum concentrations of campesterol and sitosterol, a significant reduction did occur during sitostanol therapy (-47% and -51%, respectively; p < 0.01). Fecal excretion of neutral sterols increased from 6.7 mg/kg per day during the control period to 9.7 mg/kg per day during sitosterol administration (p < 0.05), and to 12.6 mg/kg per day during sitostanol administration (p < 0.05 compared with diet and sitosterol periods), indicating an increase in the inhibition of intestinal cholesterol absorption. All children completed the study and no obvious side effects occurred. The data indicate that sitostanol, even with a dose four-fold lower than that of sitosterol, was significantly more effective in reducing elevated levels of low-density lipoprotein cholesterol, and the reduction in serum lipid levels was of the same magnitude as that observed with systemic lipid-lowering drugs. These results suggest that sitostanol, a nonabsorbable plant sterol, could be the drug of choice for treating familial hypercholesterolemia in childhood. PMID- 8429450 TI - Pharmacokinetics and hematologic response to subcutaneous administration of recombinant human erythropoietin in children undergoing long-term peritoneal dialysis: a multicenter study. AB - For a study of the pharmacokinetics and hematologic response of subcutaneously administered recombinant human erythropoietin (rHuEPO), 24 children (mean age, 10 years 3 months; range, 3 months to 18 years) maintained by peritoneal dialysis and with anemia caused by end-stage renal failure (mean hemoglobin level, 6.5 gm/dl; range, 4.7 to 7.9) were treated with rHuEPO administered subcutaneously at an initial dose of 25 IU/kg twice per week. After a 4-week interval, in the case of no response (hemoglobin increase < or = 1 to 1.5 gm/dl per month) the rHuEPO dosage was increased every 4 weeks according to the following schedule: 50, 75, 100, and 150 IU/kg twice per week. The administration of rHuEPO produced a rapid increase in serum concentration with a mean peak level of 59.8 mU/ml after 9 hours. Mean area under the curve to 72 hours was 2020 mU/ml per hour (range, 568 to 6609); mean elimination half-life and mean residence time were, respectively, 25.2 hours (range, 6.2 to 58.7) and 42.0 hours (range, 10.9 to 96). Of 24 children entered in the study, six had the drug suspended early because of renal transplantation (n = 1), lack of compliance (n = 4), or severe worsening of hypertension (n = 1). Eighteen patients had increased hemoglobin levels (to 9.4 +/- 1.7 gm/dl after 24 weeks of treatment). No correlation was found between the increase in hemoglobin concentration and any of the pharmacokinetic data or the peak erythropoietin level reached during the kinetic profile. Eight children required an increase of antihypertensive medications to maintain satisfactory blood pressure values. We conclude that low doses of subcutaneously administered rHuEPO slowly release the drug into the blood and satisfactorily increase hemoglobin levels with very few side effects. PMID- 8429451 TI - Outpatient management with oral corticosteroid therapy for obstructive conditions in chronic granulomatous disease. AB - We report the outpatient management of three patients with X-linked chronic granulomatous disease, two of whom had episodes of gastric outlet obstruction and another, urinary bladder obstruction. These obstructive conditions were successfully treated with 2-week courses of orally administered corticosteroids with or without the addition of orally administered clindamycin. There were no infectious or other adverse reactions. PMID- 8429452 TI - Maltreatment of children born to teenage mothers: a comparison between the 1960s and 1980s. AB - OBJECTIVE: To determine whether the rates or the reporting of maltreatment of children born to teenage mothers changed from the 1960s to the 1980s. DESIGN: Comparison of two cohorts. SETTING: Ambulatory services of Yale-New Haven Hospital, New Haven, Conn. PATIENTS: Consecutive children born to black teenage mothers; the "early" cohort comprised 148 children born from September 1967 through June 1969, and the "late" cohort, 159 children born from October 1979 through December 1981. OUTCOME MEASURES: Each injury for which the child was medically evaluated from birth to 5 years of age was categorized by predefined criteria as follows: maltreatment (abuse, neglect, or sexual abuse), household violence, unintentional injury-neglect, unintentional injury, or insufficient information. RESULTS: The rates of maltreatment in the early cohort and in the late cohort were similar: 12.8% versus 10.7% (relative risk (RR) = 0.83; 95% confidence interval (CI) = 0.45, 1.54). The rates of unintentional injury also were similar: 52% in the early cohort versus 60% in the late cohort (RR = 1.17; 95% CI = 0.96, 1.43). In contrast, 30% of the episodes of maltreatment were reported to the state protective service agency in the early cohort versus 65% of episodes in the late cohort (RR = 2.14; 95% CI = 1.08, 4.26). CONCLUSION: Among children born to teenage, black, inner-city women, the rates of child maltreatment are similar from the late 1960s to the 1980s. The increased rates of reporting of maltreatment reflect increased rates of recognition by clinicians, rather than true increases in prevalence. PMID- 8429453 TI - Unforgettable patients. PMID- 8429454 TI - Unforgettable patients. PMID- 8429455 TI - Dextromethorphan in infantile nonketotic hyperglycinemia. PMID- 8429456 TI - Assessing the outcome of pediatric intensive care. PMID- 8429457 TI - Timing of renal ultrasound studies in neonates. PMID- 8429458 TI - Incidence of acute rheumatic fever. PMID- 8429459 TI - Growth hormone therapy in nephropathic cystinosis. PMID- 8429460 TI - Hormonal responses to surgical stress in children. AB - The hormonal responses to surgical stress in adults are well characterized. We hypothesized that children have age-related differences in the "stress responses" to surgery. To test this hypothesis we prospectively studied 98 children (aged 2 to 20 years) undergoing elective surgical procedures under general anesthesia. Preoperative and postoperative (1 hour postoperation) blood samples were obtained and serum prolactin and cortisol concentrations were measured. Patient data were stratified by patient age and length of operation. All patients had significant (P < .05) increases in serum cortisol and prolactin concentrations 1 hour postoperatively as compared with preoperative values. However, there were no significant differences in prolactin and cortisol responses to surgery based on the age, anesthetic technique, or length of operation. Females had higher mean (+/- SD) serum prolactin concentrations (78.41 +/- 62.23 micrograms/L) as compared with males postoperatively (39.8 +/- 21.75 micrograms/L) (P < .05). We conclude the following: (1) children have significant increases in circulating prolactin and cortisol concentrations following surgery and anesthesia, and that those increases are not affected by age, length of surgery, or anesthetic technique; and (2) females have greater prolactin responses to surgery and anesthesia than males. PMID- 8429461 TI - Jejunum is preferable for construction of a Bianchi bowel-lengthening procedure in swine short bowel. AB - This study compared the efficacy of a Bianchi bowel-lengthening procedure performed in residual ileum and jejunum of a 75% short bowel model. Eighteen female piglets underwent a 75% mid small bowel resection. After a 6-week period, animal weights were similar and pigs were randomly assigned to one of three treatment groups: (1) a control group receiving no further therapy; (2) a group receiving a Bianchi procedure in the residual jejunal segment; and (3) a group receiving a Bianchi procedure in the residual ileal segment. All were followed for a further 12 weeks. Jejunal Bianchi-treated short bowel animals demonstrated a greater final weight gain (78.8 +/- 4.9 kg) compared with nontreated short bowel (63.0 +/- 6.6 kg) and ileal Bianchi-treated short bowel groups (69.3 +/- 6.9 kg) in addition to a larger jejunal diameter. The increased weight gain in the jejunal Bianchi-treated group was not a consequence of initial bowel length, food intake, changes in bowel length, digestibility of nitrogen or fat, or nutritional status. Furthermore, kinetic constants for D-glucose absorption following 18 weeks of short-bowel syndrome demonstrated a lowered glucose maximal transport rate (Vmax) in animals with nontreated short bowel compared with sham operated controls. Additionally, jejunal and ileal glucose Vmax was further lowered in the presence of a Bianchi procedure. We conclude that: (1) during short-bowel syndrome, body weight gain was significantly higher in animals when the Bianchi procedure was performed in jejunum; (2) the short-bowel syndrome decreased intestinal glucose absorption; and (3) the Bianchi procedure itself further impaired glucose transport. PMID- 8429462 TI - Respiratory gas exchange in response to fat-free parenteral nutrition: a comparison after thoracic or abdominal surgery in newborn infants. AB - Thoracic surgery is known to cause a postoperative respiratory failure because of the mechanical problems following chest wall disruption and/or diaphragmatic dysfunction. This study was to verify whether the fat-free intravenous nutritional support of neonates who underwent thoracic surgery could lead to a CO2 production exceeding the patients' respiratory reserves. Respiratory gas exchange and alveolar ventilation were obtained by indirect calorimetry and continuous recordings of transcutaneous PO2 and PCO2. These noninvasive measurements were compared at the same age of 7 +/- 1 days between a group of 7 newborn infants (mean +/- SEM: 3.09 +/- 0.14 kg, 39 +/- 1 weeks) after thoracic surgery versus a group of 8 newborn infants (2.88 +/- 0.17 kg, 37 +/- 1 weeks) after abdominal surgery. The intravenous macronutrient support was the same between both groups: 14 g/kg/d of glucose, 2 g/kg/d of amino acids, 250 kJ/kg/d of energy. One week after surgery, the global metabolic rate (195 kJ/kg/d) was not increased, and comparable between both groups. We documented that early after thoracic surgery, the ventilatory compensation required to handle the CO2 production (6.7 +/- 0.2 mL/kg/min) associated with a positive energy balance (45 +/- 8 kJ/kg/d) was effective. PMID- 8429463 TI - Ileocecal resection in neonates and infants: a follow-up study. AB - From 1977 to 1983 ileocecal resection was done in five neonates and three infants who were admitted to our pediatric surgical units. Their growth, hematology, and serum biochemistry were examined and compared with that of an ileal resection group without ileocecal resection. The body weight and height of all patients of the ileocecal resection group were within normal ranges. All patients undergoing ileocecal resection in neonates had moderate diarrhea but condition of fecal evacuation improved after age 6. None of the control ileal resection group had diarrhea since age 2. No significant differences were noted in hematology and serum biochemistry (protein metabolism, lipid metabolism, bile acid, and vitamin B12) data between the ileocecal resection groups and the control group. Our findings show that after ileocecal resection without extensive ileal resection in neonates and infants, adequate nutritional status can be maintained. PMID- 8429464 TI - Early hepatobiliary dysfunction during total parenteral nutrition: an experimental study. AB - The etiology and pathophysiology of the liver disease associated with total parenteral nutrition (TPN) are unknown. In this study, we have attempted to define the early changes in hepatobiliary function during TPN in young rabbits nourished totally by the intravenous route for 3, 5, and 15 days, with age matched rabbits on lab chow serving as controls. A decrease in basal bile flow along with elevations of serum bile acids and cholesterol was seen. The capacity for biliary secretion of sulfobromophthalein and of ursodeoxycholic acid was measured at the end of each diet regimen. Early impairment of biliary sulfobromophthalein (BSP) secretion was seen after 5 days of TPN, with no further deterioration after 15 days. Maximal bile acid secretory rate and bile flow, in response to the ursodeoxycholic acid load, were decreased after 15 days of TPN. Furthermore, after 15 days of TPN, both the volume of gallbladder bile and its bile acid content increased. The combined effects of the enteral fast and the intravenous administration of all nutrients were bile acid sequestration in an adynamic gallbladder with interruption of the enterohepatic circulation. In the parenterally fed rabbit, we have demonstrated bile secretory failure and gallbladder sludge, the two common complications of clinical TPN. These may be the early events that subsequently lead to cholestasis and liver damage in neonates maintained on prolonged TPN. PMID- 8429465 TI - Audit filters in quality assurance in pediatric trauma care. AB - Quality assurance (QA) systems use audit filters to help identify not only deaths and medical complications, but also cases that reflect deficiencies in the timeliness or appropriateness of care. Through our trauma center QA process, we studied three groups of audit screens for trauma care: two set forth by the Committee on Trauma of the American College of Surgeons, a minimum set of 12 audit filters proposed in 1987 and an expanded version recently proposed in 1990 (studied retrospectively), and one set of filters already in use in our system. A peer review committee determined whether deaths and complication were preventable, and judged the timeliness and appropriateness of care. From July 1, 1989, through June 30, 1990, 844 admitted trauma patients came under QA review. During this period 13 (1.5%) died; 35 (4.1%) suffered at least one medical complication. 140 children (16.6%) violated one of the 12 minimum audit filters suggested by the Committee on Trauma (which includes deaths and medical complications), one of the additional filters used by the Trauma Program QA system, or both. Ninety patients (10.6%) failed one of the 12 minimum audit filters; 104 (12.3%) failed one of the additional filters used by the Trauma Program QA system. Filters that involved medical management issues (late operations, return to operating room, airway, failed reduction, infections, missed injuries, readmissions to intensive care unit, return to the emergency department) frequently involved aspects of inappropriate care (72.5%, 37/51 violations) and were associated with actual deaths or medical complications (52.9%, 27/51).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429466 TI - Wound infection in pediatric surgery: a study in 1,094 neonates. AB - In an analysis of 1,433 wounds created in 1,094 neonates admitted to a regional neonatal surgical unit during the period April 1975 to December 1987, the mean incidence of infection was 16.6%. During this time there was an increase in the incidence of infection from 12.5% in the first 6 years to 18.8% in the last 7 years (P < .01). Contaminated wounds had an infection rate of 20.7%, whereas the rate in clean wounds was 11.1% (P < .001). Gestational age and birth weight had no influence on the incidence of wound infection. Increasing wound length (P < .001), increasing duration of operation (P < .001), and contamination at operation (P < .001) were all associated with a higher incidence of infection. Staphylococcal species were the most frequently isolated organisms from all categories of infected wounds (clean, potential, and actual contamination). PMID- 8429467 TI - Silastic catheterization of the axillary vein in neonates: an alternative to the internal jugular vein. AB - The axillary vein is a suitable alternative to the jugular venous system for tunnelled silastic catheterization in neonates, and should be included in the armamentarium of the surgeon who treats neonates. It is technically easy and is comparable to the internal jugular vein in terms of complications. Proper positioning of the catheter tip can sometimes be problematic, but without a resultant increase in morbidity or mortality. PMID- 8429468 TI - Prenatal management of congenital cystic adenomatoid malformation of the lung. AB - In utero evolution and postnatal outcome were studied for 18 cases of congenital adenomatoid malformation (CAM) diagnosed by prenatal ultrasound. Five were macrocystic CAM, 9 were microcystic, and 4 were homogeneously hyperechogenic. Three fetuses presented with hydrops. Pulmonary amniotic shunting was performed in 3 patients. Outcome was the following: 4 were aborted, 1 died neonatally, and 13 survived. Four of these infants required no surgery in the neonatal or postneonatal period. In three of these, the size of the mass had decreased spontaneously in utero. Outcome did not appear to be related to the anatomic type of CAM nor to the presence of moderate polyhydramnios, but was related to the degree of mediastinal compression and to the existence of hydrops. A clearer understanding of the natural evolution of CAM is useful to determine the indications for in utero therapy. PMID- 8429469 TI - The long-term results of diaphragmatic plication. AB - Twelve patients who had undergone diaphragmatic plication with the diagnosis of congenital diaphragmatic eventration between 1975 to 1989 were evaluated for the long-term results of plication. Assessment of the long-term functions of the plicated diaphragms 1.5 to 11 years postoperatively was achieved by fluoroscopic, ultrasonographic, and spirometric studies. The absence of paradoxical motion with normal localization of the diaphragms in all patients, and satisfactory motions of diaphragms in 9 patients were documented by fluoroscopy. Measurements of diaphragmatic thicknesses showed that plicated diaphragms of all patients maintained their growths in proportion to the contralateral sides. Additionally, normal values of pulmonary function tests in five of six patients of suitable age for spirometry were obtained. All the clinical studies demonstrated that diaphragmatic plication did not interfere with further development of diaphragms, and late functional results of the plication were acceptable. This supports the choice of surgery in the treatment of diaphragmatic eventration. PMID- 8429470 TI - Prosthetic diaphragm reconstruction in the growing animal. AB - Widespread use of extracorporeal membrane oxygenation (ECMO) has allowed an increasing number of infants with total agenesis of the diaphragm to survive. Polytetrafluoroethylene (PTFE) is the most widely used material for reconstruction. However, recurrent hernia is a growing problem; PTFE also does not grow with the patient. This study evaluated different materials for diaphragmatic reconstruction in growing animals. Sprague-Dawley rats with a mean weight of 93 g were anesthetized and underwent laparotomy. The control group had an incision into the diaphragm with primary repair. The other three groups underwent complete removal of the left hemidiaphragm and were randomly assigned to one of three reconstruction methods: oxidized cellulose, polyglactin mesh, or a 1-mm PTFE patch. All patch materials were sewn around the ribs circumferentially and into the membranous portion of the central diaphragm medially with 4-0 silk. Thirty-seven animals survived operation, were followed with weekly chest radiographs, and were killed when they reached 400 g. The radiographs were reviewed in a blinded fashion by two observers as were the necropsies, and rib deformity was graded on a scale of 0 to 3. Histological examination of several animals from each group was performed. There was significantly greater rib deformity (2.0 v 0.2, P < .01) in the PTFE group versus controls with 5 of 10 animals also having a smaller thorax. The PTFE pulled away from the chest wall in the animals leaving a fibrous remnant anteriorly. The polyglactin group had significantly more animals with eventration (P < .03, 7/10) compared with all others.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429471 TI - Incidence and management of gastroesophageal reflux after repair of congenital diaphragmatic hernia. AB - During a period of 6 years, 31 of 54 infants with congenital diaphragmatic hernia survived and were evaluated to study incidence and management of gastroesophageal reflux. At 6 months' follow-up 16 patients had gastroesophageal reflux proven by upper gastrointestinal series; at 12 months' follow-up 11 patients. Three patients having gastroesophageal reflux did not respond to medical treatment and underwent Nissen fundoplication. We conclude that after successful treatment, congenital diaphragmatic hernia is likely to be complicated by gastroesophageal reflux. We could not define a predictive feature for gastroesophageal reflux in the individual patient surviving congenital diaphragmatic hernia. PMID- 8429472 TI - Proteus syndrome: diagnosis and surgical management. AB - The congenital hamartomatous syndrome known as the "Proteus syndrome" (PS) manifests itself with regional giantism, lymphangiomatous hamartomas, and other variable features. Review of the medical literature shows approximately 50 cases reported to date. Since this syndrome has only recently been defined, the management of these patients has been speculative and often children are not treated. This report summarizes the characteristics of the PS and presents eight additional cases. All of the eight children had regional giantism with macrodactyly and skeletal hypertrophy. Asymmetrical leg length was pronounced in five children. All children had large lymphangiomas, the majority of which involved the trunk. Three of the children have been followed through adolescence, two into late childhood, and three into early childhood. In contrast to previous reports, we believe that early surgical reconstruction is necessary to reduce deformities due to the giantism and the large hamartomas. During extensive excisions, residual abnormal tissue is often needed in the reconstruction and it is not unusual for postoperative leakage of lymph to be prolonged. All of the children in this series have benefited both physically and emotionally from extensive surgical reconstruction. PMID- 8429473 TI - Efficacy of the Nissen fundoplication in the management of gastroesophageal reflux following esophageal atresia repair. AB - From January 1974 to December 1988, 80 patients with esophageal atresia (EA) and tracheoesophageal fistula (TEF) were treated at the C.S. Mott Children's Hospital with division of their TEF and esophagoesophagostomy. Thirty-four in this group developed gastroesophageal reflux (GER). After an unsuccessful trial of medical management, 21 underwent Nissen fundoplication, and an additional child with refractory GER died intraoperatively before completion of her fundoplication. Following fundoplication, only eight patients had an uncomplicated course with elimination of reflux and no postoperative dysphagia. Wrap disruption and recurrent reflux occurred in 7 of the 21 (33%), a markedly higher incidence than the 10% figure seen in 220 children without EA who have undergone fundoplication at this institution. Upward tension on the wrap due to the presence of a shortened esophagus probably predisposes to an increased frequency of fundoplication failure in the EA child. In addition, postoperative dysphagia requiring prolonged gastrostomy feedings complicated eight otherwise successful initial or redo-fundoplications. Prolonged dysphagia in this group is likely related to the inability of the dyskinetic esophagus, common in EA, to overcome the increased resistance caused by the Nissen fundoplication. Three deaths (14%) from complications related to antireflux surgery occurred in the series. Although 15 of the 21 children (71%) eventually had excellent long-term results following initial or redo-fundoplication with elimination of reflux and normalization of oral intake, morbidity and mortality were clearly significant. Due to the high incidence of postoperative dysphagia and recurrent reflux, the transabdominal Nissen fundoplication may not be appropriate in EA patients. PMID- 8429474 TI - The effect of tube feeding on postprandial gastroesophageal reflux. AB - Distal esophageal pH in supine position was monitored for 6 hours in 13 asymptomatic infants. During this period the patients received 15 to 20 mL/kg of 5% glucose solution twice every 3 hours. Each meal was fed orally or through a pH sensor tube. When the patient was fed through a tube, the meal was introduced into the stomach or into the distal esophagus. The extent of reflux during 2 hours after feeding was compared between oral and tube feeding. Comparing oral and intragastric tube feeding, intragastric tube feeding showed significantly fewer reflux events, a smaller percentage of time in the esophageal pH below 4, and a shorter duration of the longest single reflux event. Comparing oral and intraesophageal tube feeding, intraesophageal tube feeding showed the same pattern. The data indicated that tube feeding significantly diminishes the postprandial reflux. PMID- 8429475 TI - Esophagitis in infants with hypertrophic pyloric stenosis: a source of hematemesis. AB - This study was undertaken to clarify the source of blood in the vomitus of patients with hypertrophic pyloric stenosis (HPS). Twenty-one infants with HPS were examined. Hematemesis was noted in 14 infants. Esophagogastric endoscopy showed a 100% incidence of esophagitis and in one patient gastric erosion was also observed. Histological study of the esophageal mucosa showed evidence of esophagitis in 18 patients (85.7%). Preoperative pH monitoring showed gastroesophageal reflux (GER) in all infants. Excessive acid exposure (> or = 7%) was significantly correlated with the grade of esophagitis and the incidence of hematemesis, whereas acid exposure time was shorter in the cases without histological esophagitis. These results suggested that the source of bleeding in HPS is the esophageal mucosa affected by esophagitis secondary to excessive acid reflux. Although there is obvious massive gastroesophageal reflux in HPS, it is too difficult to evaluate the lower esophageal sphincter function in HPS. PMID- 8429476 TI - Hepatic histology and the development of esophageal varices in biliary atresia. AB - Histological features in liver biopsy specimens taken from 71 infants at the time of surgery for biliary atresia (portoenterostomy) were analyzed using a scoring system and compared with an endoscopic grading of esophageal varices performed at a mean age of 3.4 years. The analysis showed no correlation between a "global" score, which represented the severity of all histological changes in the original biopsy specimen, and the severity of esophageal varices. Further analysis also showed no correlation with any individual histological feature (eg, fibrosis). These findings failed to confirm a previous study, which suggested a relationship between liver changes at surgery and the later development of esophageal varices in children with biliary atresia. PMID- 8429477 TI - Perioperative management for cholecystectomy in sickle cell disease. AB - Perioperative complications of surgical procedures are frequently encountered in patients with sickle cell disease. We have reviewed our series of patients with hemoglobinopathies who underwent cholecystectomy from 1978 to 1991 to evaluate their perioperative management and clinical outcome. Twenty-two children with major sickle hemoglobinopathy underwent cholecystectomy for symptomatic cholelithiasis. All 22 were transfused to achieve a hemoglobin (Hgb) level greater than 9 g/dL and hemoglobin S (HbS) less than 37%. Fourteen underwent immediate preoperative automated red cell exchange (ARCE). The median preexchange Hgb of these patients was 8.1 g/dL (range, 6.8 to 10.5). Their median HbS was 84% (range, 53% to 97%). These patients underwent placement of an apheresis catheter under local anesthesia followed by red cell exchange. The median volume of packed red blood cells (PRBC) exchanged was 28.1 mL/kg (range, 13.8 to 58.7). The median HbS after exchange was 21% (range, 16% to 37%) and the median Hgb was 10.6 g/dL (range, 6.5 to 16.7). Eight other patients underwent sequential transfusion (3 after an exchange for an acute pulmonary vasoocclusive crisis). These patients had been prepared over an interval of 2 to 8 weeks preoperatively and had received a median of 26.9 mL PRBC/kg (range, 12.8 to 95). Following sequential transfusion the median Hgb was 11.8 g/dL (range, 9 to 15.7) and the median HbS was 19% (range, 5% to 32%) at the time of surgery. All patients received extended antigen matched blood. Complications of preoperative transfusion were minor and included two febrile-/allergic reactions and one mild superficial catheter induced phlebitis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429478 TI - Effective control of chylous ascites: an alternative approach. AB - Complete control of chylous ascites was achieved by creating a circuit for extracorporeal recirculation of the ascites fluid from the peritoneal cavity to the superior vena cava. A hemofilter was incorporated into the circuit to remove water and electrolytes while returning protein, fat, and white cells. This avoided the associated loss of calories. This system could be applied to other forms of refractory ascites. PMID- 8429479 TI - The place of ultrasonographic examination in the initial evaluation of children sustaining blunt abdominal trauma. AB - Physical examination is stated to be inaccurate in detecting organ injury after blunt abdominal trauma (BAT) in children and the use of diagnostic peritoneal lavage (DPL) and computed tomography (CT) are suggested. However, assessments for the need of such diagnostic aids are quite subjective. The records of 109 patients initially evaluated by ultrasonography (US) for BAT were reviewed to determine whether an easily performed, quick method such as US could be used for selection. US showed free intraperitoneal fluid (FIF) in 30 patients (27.5%) and retroperitoneal injury in 5 patients (4.5%). Of the 30 patients with FIF, 23 patients (76.7%) were treated conservatively but 7 (23.3%) required laparotomy. The correlation between the amount of FIF and the requirement for operative treatment was statistically significant (P < .01). Of the 74 patients (68%) without FIF, clinical outcome was uneventful in 72 (97.3%) while 2 patients (2.7%) required laparotomy for peritonitis and ileal perforations were encountered. The present study has showed that US is inaccurate in detecting solid intraabdominal injuries; however, it is reliable in detecting FIF produced as a result of intraabdominal organ injuries and retroperitoneal organ injuries. We suggest the use of US as the objective initial evaluation method of choice on a routine basis. PMID- 8429480 TI - Magnetic resonance imaging of abdominal masses in children. AB - Seventeen children with abdominal masses (16 solid and 1 complex cystic) were evaluated by magnetic resonance imaging (MRI). Computed tomography (CT) was also performed in 12 cases. MRI findings were correlated with CT and histopathologic findings. In 10 cases MRI provided more specific diagnostic information than CT. In 2 cases MRI did not provide any significant additional information. In 5 cases, MRI provided enough specific information alone that CT was not deemed necessary prior to clinical management. We conclude from our experience that MRI should be the modality of choice in the further evaluation of noncystic masses after initial sonography. MRI manifestations play a major role in establishing the nature and source of abdominal masses which directly affect their clinical management. Compared with CT, MRI was better in localizing and determining the extent of lesion in its primary and secondary sites, without the need for iodinated contrast injection. PMID- 8429481 TI - Congenital duodenal obstruction: a 32-year review. AB - Although survival in infants with congenital intestinal obstruction has improved, duodenal obstruction continues to present unique challenges. One hundred thirty eight newborns and infants (aged 0 to 30 days) were treated for congenital duodenal obstruction. Sixty-five were boys and 73 were girls. Sixty-one (45%) were premature. Forty-six had an intrinsic defect (atresia, web, stenosis, or duplication), 64 had an extrinsic defect (annular pancreas or malrotation with congenital bands), while 28 had various combinations of these. Presenting signs included vomiting (90%, bilious in 66%), abdominal distention (25%), dehydration (24%), and weight loss (17%). Although plain film abdominal x-ray was diagnostic in 58%, upper and/or lower gastrointestinal contrast studies were obtained in 71% of infants to confirm diagnosis. Thirty-eight percent of patients had associated anomalies, including Down's syndrome (11%), cardiac defects, other atresia, other trisomy syndrome, imperforate anus, and central nervous system anomalies. Fourteen patients (10%) had 3 or more other anomalies, many of which required additional surgical therapy. The operative repair of the various defects included Ladd's procedure for malrotation (31%), duodenoduodenostomy (14%), duodenojejunostomy (22%), gastrojejunostomy or gastroduodenostomy (4%), excision of the web and duodenoplasty (3%), or combination of the above (22%). Gastrostomy was placed in 61%. One hundred twenty-eight patients survived (93%). The causes of death were combinations of sepsis, pneumonia, brain hemorrhage, short bowel, and cardiac anomaly. Eight of 10 (80%) who died had other serious anomalies. Twenty patients (14%) required reoperation 5 days to 4 years postoperatively for obstructing lesions (5), wound dehiscence (3), anastomotic leak or dysfunction (6), other atresias (2), choledochal cyst (1), pyloric stenosis (1), and gastroesophageal reflux (2).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429482 TI - Malrotation of bowel: variable patterns with different surgical considerations. AB - The records of 53 patients with varied roentgenographic patterns of malrotation were reviewed. The patterns were categorized as A-Infantile (Al), A-Adult (Aa), B, C, and D. The differences in form were obvious and each differed clinically, radiographically, and surgically. The differences allowed the development of prognostic insights simply based on the presence and/or position of the ligament of Treitz and the cecum as shown by barium gastrointestinal examination. The literature was reviewed to find supportive or contradictory case material or other data that complemented or devalued the developed system for determining prognosis of patients with varied bowel patterns. The developed prognostic scheme seems to satisfy the requirements of reliability. PMID- 8429483 TI - In the search for new anticancer drugs. XXIV: Synthesis and anticancer activity of amino acids and dipeptides containing the 2-chloroethyl- and [N'-(2 chloroethyl)-N'-nitroso]-aminocarbonyl groups. AB - A series of L,L- (42, 44, 46, and 60) and D,D- (43, 45, 47, and 61) dipeptide derivatives composed of phenylglycine, phenylalanine, homophenylalanine, and valine and containing a 2-chloroethylamino group at the C-terminus and an N'-(2 chloroethyl)-N'-nitroso-aminocarbonyl group at the N-terminus of the dipeptides were prepared. The dipeptide derivatives (42-47, 60, and 61) were first evaluated in vivo for their anticancer activities against the murine lymphocytic leukemia P388. Compounds 42, 44, 46, and 60 possessed activities ranging from 46 to 111 percent increase in life span (%ILS), whereas 43 was marginal (%ILS = 31) and 45, 47, and 61 were inactive. In general, the L,L-series exhibited low to good activity (%ILS = 46-111), whereas the corresponding D, D-series, except for 43 (%ILS = 31), was devoid of activity. The analogously structured monoamino acid derivatives of L-alanine (74), L-phenylalanine (75), and L-aspartic acid (76) exhibited higher activity against P388 than the dipeptide derivatives (i.e., 481, 297, and 481 %ILS, respectively). The more active representatives of dipeptides (i.e., 42, 44, and 60) and the amino acids derivatives 74-76 were then tested in vivo against the murine lymphoid leukemia L1210. Compounds 42, 44, and 60 exhibited either low or marginal activity (i.e., the %ILS values were 46, 31, and 26, respectively). Compounds 74, 75, and 76 possessed low to moderate activity, as evidenced by the %ILS values of 56, 48, and 64, respectively. The %ILS parameters obtained against the P388 and L1210 tumor lines were correlated with the corresponding lipophilicities, and there is a trend towards higher activity with concomitant decrease in hydrophobicity. PMID- 8429484 TI - gamma-Pyrone compounds. IV: Synthesis and antiplatelet effects of mono- and dioxygenated xanthones and xanthonoxypropanolamine. AB - Xanthodilol, mono- and dioxygenated xanthones, and 1,3-, 2,3-, 3,4-, 3,5-, 1,6-, 2,6- and 3,6-dioxygenated xanthones were synthesized from benzophenone precursors by Friedel-Crafts acylation and subsequent base-catalyzed cyclization to eliminate methanol. 3-Hydroxy-xanthone, xanthodilol, 2,3-dihydroxyxanthone diacetate, and 3,4-dihydroxyxanthone and its diacetate showed potent antiplatelet effects on arachidonate- and collagen-induced aggregation. 3,5-Dihydroxyxanthone and its diacetate, 1,6-dimethoxyxanthone, and 3,6-dihydroxyxanthone and its diacetate showed potent antiplatelet effects on arachidonate-induced aggregation. PMID- 8429485 TI - Potentiometric determination of the thermodynamic ionization constants of deferoxamine. PMID- 8429486 TI - Cutaneous bioavailability in hairless rats of tretinoin in liposomes or gel. AB - Topical bioavailability of drugs incorporated in liposomes is not well known. We compared the skin penetration of tretinoin in liposomes and in a classical alcoholic gel. [3H]Phosphatidylcholine dipalmitoyl (DPPC) and [14C]tretinoin (0.14%) were incorporated in the phospholipidic phase of the liposomes, and [14C]tretinoin was incorporated in a gel for comparison. Skin absorption was studied in vitro with Franz cells. In vivo distribution in cutaneous structures was studied according to Schaefer's method. Liposomes impregnated the stratum corneum, with a partial dissociation between tretinoin and phosphatidyl-choline dipalmitoyl. In dermis, tretinoin diffused alone. Tretinoin release seemed to be controlled, and steady state was reached later with liposomes than with gel. This phenomenon was linked with a significantly reduced absorption (1.60% for liposomes versus 3.09% for the gel) and higher retention in epidermis (mainly stratum corneum) and dermis (41 and 13%, respectively, with liposomal form versus 18 and 8%, respectively, with gel form). This study clearly shows that, compared with the gel, the liposome formulation tends to improve the local effect of tretinoin in the skin and decrease the systemic absorption. PMID- 8429487 TI - Membrane transport of tetraphenylphosphonium and its homologues through the planar phospholipid bilayer: concentration dependence and mutually competitive inhibition in membrane passive transport. AB - The concentration dependence and mutually competitive inhibition in the membrane transport of the lipophilic cations tetraphenylphosphonium (TPP+) and its homologues were studied by use of a planar phospholipid bilayer. The current voltage characteristics were analyzed by use of a model developed by Ketterer et al. (J. Membr. Biol. 1971, 5, 225-245). The conductance at 0 mV [G(0)], which represents the transport ability, increased with an increase in concentration, and the G(0) values revealed saturation at higher concentrations. The relationship between G(0) and concentration was well fitted to a Michealis-Menten type equation with a saturable component (maximum conductance [G(0)max] and concentration yielding one-half of G(0)max[Km]). The G(0)max and Km values were calculated to be 4.1 x 10(-8)-242 x 10(-8) S/cm2 and 13-489 microM, respectively, depending on the lipophilic cations used. The G(0) values normalized by the corresponding G(0)max values were plotted against the concentrations normalized by the corresponding Km values. Normalized curves for TPP+ homologues were all superimposed on a single curve. The mechanism for the saturation of G(0) values may involve the adsorption of TPP+ homologues to a planar phospholipid bilayer, because the Km values calculated in the present study were comparable to those for adsorption to phosphatidylcholine liposomes. We further determined the mutual inhibition of membrane transport by these homologues. Tetraphenylmethylphosphonium at 1 mM, a concentration threefold higher than the Km value for this compound, reduced the G(0) for other TPP+ homologues by 38-55%, whereas reduction by 10 mM tetraethylammonium was minimal. The type of inhibition was classified as mutually competitive.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429488 TI - Hygroscopic stability and dissolution properties of spray-dried solid dispersions of furosemide with Eudragit. AB - Spray-dried solid dispersions of furosemide-Eudragit RS100 and RL100 (ethyl methacrylate chlorotrimethylammoniumethyl methacrylate copolymer) were studied to determine their stability at 45 degrees C and 0, 44, and 75% relative humidity and their dissolution characteristics. The crystallization rates of the solid dispersions were measured by X-ray powder diffractometry and calculated on the basis of the Jander equation to estimate physicochemical stability in the presence of water vapor. The stability of the solid dispersion depended on the kind of acrylic resins and the drug content. The solid dispersion with a drug: Eudragit RL100 ratio of 1:3 was the most stable at 75% relative humidity. The dissolution profiles of the solid dispersions were measured in a pH 6.8 buffer at 37 degrees C. The equilibrium drug concentration of the solid dispersions were estimated from the dissolution profiles after 24 h. The equilibrium drug concentration decreased with an increasing Eudragit quaternary ammonium group: furosemide molar ratio. The equilibrium drug concentrations of the solid dispersions with > 0.3 mol of the quaternary ammonium group were approximately 0. Thus, the drug was completely adsorbed on Eudragit. PMID- 8429489 TI - Determination of m-nitrophenol and nipecotic acid in mouse tissues by high performance liquid chromatography after administration of the anticonvulsant m nitrophenyl-3-piperidinecarboxylate hydrochloride. AB - The nipecotic acid ester m-nitrophenyl-3-piperidinecarboxylate (MNPC) possesses anticonvulsant activity. In the present study, the metabolites m-nitrophenol and nipecotic acid were determined in mouse blood and brain tissue after administration of MNPC. This determination was used as an indication of the distribution of the parent compound MNPC and to provide information regarding the differences in distribution between the enantiomers of MNPC, the times of onset, and effectiveness when (+/-) MNPC was administered by subcutaneous (sc) and intraperitoneal (ip) routes. m-Nitrophenol was determined by a previously reported high-performance liquid chromatography (HPLC) method. There was no significant difference in m-nitrophenol distribution after sc administration of (+)MNPC and (-)MNPC (400 mg/kg each). This similar pattern of distribution is in agreement with the earlier reported equi-effectiveness of the enantiomers of anticonvulsants. Peak m-nitrophenol levels in blood, which occurred at 15 min, were three times greater when (+/-) MNPC was administered by ip injection as compared with sc injection. This significant difference is most likely due to enhanced absorption and the peripheral metabolism of MNPC by the liver when the ip route is employed. A novel HPLC assay for the determination of nipecotic acid in mouse brain was developed, based on a modification of a reported amino acid analysis procedure. The results of the brain distribution studies showed that nipecotic acid brain levels peaked at 30 min after sc administration of ( )MNPC.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429490 TI - Determination of free bile acids in pharmaceutical preparations by packed column supercritical fluid chromatography. AB - A method was developed for the baseline separation of common free bile acids by supercritical fluid chromatography. A phenylbonded silica column, with UV detection at 210 nm, and carbon dioxide modified with methanol as the mobile phase were used. The influence of the stationary phase, modifier concentration, temperature, column pressure, and modifier identity on retention was studied. The separation obtained is at least eight times faster than those achieved for similar mixtures by the existing chromatographic techniques. This new procedure is applicable to the assay of ursodeoxycholic acid and chenodeoxycholic acid in capsule and tablet formulations. Individual dosage forms were disintegrated in methanol and an aliquot of the resulting suspension was filtered for the supercritical fluid chromatographic analysis. The method is rapid, accurate, and reproducible. PMID- 8429491 TI - Comparison of the in vivo and in vitro nuclear magnetic resonance detection of trifluoromethyl penicillin V in rats. AB - Fluorine nuclear magnetic resonance (NMR) spectroscopy and magnetic resonance imaging were examined as noninvasive methods for characterizing antibiotic disposition and pharmacokinetics in vivo. For determination of their utility, a 19F surface coil was constructed and an m-(trifluoromethyl)-containing penicillin V analogue (LY242072; 1) was synthesized. Various concentrations of 1 were injected intravenously into anesthetized rats whose urethras were occluded. The animals were placed on the surface coil, which was tuned to 19F, and then into a 4.7-T, 33-cm bore instrument, in which in vivo measurements of 1 were made on urine excreted into the bladder. At sacrifice, the urine was collected, and antibiotic levels were determined in vitro by both HPLC and high-resolution NMR. The limit of detection of 1 by NMR was 0.7 mg/mL of urine. When compared with standard in vitro quantitative methods using current technology, quantitation by in vivo surface coil NMR is not precise. Magnetic resonance imaging was used to image the bladder at a 35-mm3 voxel resolution with datum collection times of approximately 1 h. The 19F surface coil was used successfully to spectroscopically locate xenobiotic fluorine in the rat thorax. 19F NMR may offer an opportunity for the noninvasive in vivo detection of the distribution of various classes of therapeutic compounds. PMID- 8429492 TI - Determination of ceftibuten in sputum by column-switching high-performance liquid chromatography on-line with thermospray mass spectrometry. AB - A column-switching high-performance liquid chromatographic assay on-line with thermospray mass spectrometric detection is described for the determination of ceftibuten, an oral cephalosporin antibiotic, in human sputum. The method does not require sample pretreatment and provides increased selectivity not available from the previously reported method with UV detection. The thermospray mass spectrometric detection conditions were optimized for ceftibuten. The technique has a detection limit of 0.50 micrograms/mL and allows precise, simple, and accurate determination of ceftibuten in sputum over the range 0.50-10.00 micrograms/mL. PMID- 8429493 TI - High-performance liquid chromatographic determination of total thiamine in human plasma for oral bioavailability studies. AB - A high-performance liquid chromatographic method for the determination of total thiamine in plasma was developed for a relative bioavailability comparison of two oral thiamine preparations. After separation of thiamine mono- and diphosphates with the phosphatase enzyme, the total thiamine was subjected to reversed-phase high-performance liquid chromatography, postcolumn oxidized to thiochrome with K3[Fe(CN)6], and detected by fluorescence. In the bioavailability study, 16 human subjects were put on a low-thiamine diet for 3 days. On the 2nd and 3rd days, 14 blood samples per subject and day were taken at the same times each day. Drug administration did not take place until the 3rd day. After deduction of the native concentrations of total thiamine detected on the 2nd day (mean value of approximately 7 ng/mL), the post-treatment pharmacokinetic parameters were determined (two different preparations, each with 200 mg of thiamine.HCl). PMID- 8429494 TI - Hypolipidemic effects of 6-amino-2-mercapto-5-methylpyrimidine-4-carboxylic acid in rats and tissue culture cells. AB - 6-Amino-2-mercapto-5-methylpyrimidine-4-carboxylic acid (AMMPCA) was found to be a potent hypolipidemic agent at oral doses of 10 and 20 mg/kg/day in rodents. This agent was observed to affect de novo lipid synthesizing enzyme activities in a manner that resulted in lower lipid levels in tissues including the aorta wall. Very low density lipoprotein and low density lipoprotein cholesterol levels were reduced, whereas high density lipoprotein cholesterol levels were significantly increased. AMMPCA interfered with low density receptor activity, suggesting that the drug blocked lipid uptake by peripheral tissue while stimulating binding of lipid to the high density lipoprotein receptor, which should accelerate lipid clearance from the tissues and blood. A second mode of action of the drug is enhanced biliary excretion of lipids, but there was no evidence of a lithogenic effect. Acute toxicity studies in rodents support the fact that AMMPCA is safe in its therapeutic dose range. PMID- 8429495 TI - Hydrolysis of pharmaceutically relevant phosphate monoester monoanions: correlation to an established structure-reactivity relationship. AB - The kinetics of dephosphorylation of dilute aqueous solutions of 3 phosphoryloxymethyl-5,5-diphenylhydantoin (1) and estrone phosphate (2) were studied as a function of pH, buffer concentration, and temperature at an ionic strength of 0.5 M. The resulting pH-rate profiles displayed bell-shaped regions with maxima between pH 3 and 5, where the monoanionic phosphate species predominate; mechanistically, these regions involved spontaneous or water catalyzed dephosphorylation of the monoanionic phosphate species. The profile of 1 was also described with pathways involving a hydronium ion-catalyzed and a spontaneous or water-catalyzed dephosphorylation of the neutral species. The hydrolytic reactivities of the monoanionic species of the phosphomonoester prodrugs of phenytoin (i.e., 1), estrone (i.e., 2), and 2,2,2-trichloroethanol (from the literature) were well predicted by an established structure-reactivity, free-energy relationship. The correlation of these compounds to this relationship supported a common hydrolytic reaction mechanism. The small Bronsted value of 0.27 was consistent with a dephosphorylation mechanism involving a rapid transfer of the lone phosphoryl proton to the bridge-oxygen atom of the ester linkage, followed by the rate-limiting phosphorus-oxygen bond fission proceeding through a largely dissociative transition state. PMID- 8429496 TI - Complexation of moricizine with nicotinamide and evaluation of the complexation constants by various methods. AB - The solubility of the antiarrhythmic drug moricizine at physiologic pH is very low. Precipitation after rapid intravenous injection of the hydrochloride salt of moricizine could be a concern. The enhancement of solubility of moricizine near physiologic pH via complexation with nicotinamide was examined as a potential solubilization technique. The studies were performed in pH 6 and pH 7 phosphate buffers at 25 degrees C by the phase solubility method. Moricizine formed 1:1 and 1:2 complexes with nicotinamide at pHs of 6 and 7. The complexation constants K1:1 and K1:2 were estimated by a previously described scheme and equation and compared with those obtained by fitting a line and a parabola to the equations derived from the scheme for both the approximate and exact solutions. The data were best represented by a parabolic regression analysis of the exact solution of the derived equation with values for K1:1 and K1:2 at pH 6 of 16.60 and 0.93 M-1, respectively, and at pH 7 of 7.70 and 5.41 M-1, respectively. PMID- 8429497 TI - In vitro studies to investigate the reasons for the low potency of cholestyramine and colestipol. AB - The association rates, dissociation rates, and equilibrium binding of bile acids with cholestyramine and colestipol were measured under physiological conditions with the most abundant bile acids found in humans. Cholestyramine and colestipol equilibrated with the bile acids (5 mM) within 1 h and they bound > 58% and > 17% of the bile acid, respectively, when at equilibrium with physiological concentrations of bile acid (4.3-10.1 mM). However, the conjugated trihydroxy bile acids taurocholic acid and glycocholic acid dissociated rapidly from both cholestyramine and colestipol when the sequestrants, preloaded with the bile acid, were washed with the Krebs-Henseleit buffer. The taurine-conjugated and dihydroxy bile acids dissociated more slowly from cholestyramine and colestipol than the glycine-conjugated and trihydroxy bile acids and, therefore, would be expected to avoid reabsorption to a greater extent by the terminal ileum and colon in vivo. We predict from these results that the reasons for the low potency of cholestyramine and colestipol are that they bind a relatively small proportion of the trihydroxy bile acids in the duodenum and jejunum and that all of the bile acids dissociate to varying extents from the sequestrants in the terminal ileum where the unbound bile acids are reabsorbed by the gut. PMID- 8429498 TI - Simultaneous determination of propranolol or metoprolol in the presence of butyrophenones in human plasma by gas chromatography with mass spectrometry. AB - A method has been developed for the simultaneous gas chromatographic determination of propranolol or metoprolol and butyrophenones butyrophenones in human plasma in vitro. The drugs were extracted from plasma by solid-phase extraction. Calibration graphs were linear in the 50-300-ng/mL range. The recovery of the compounds was 80-90%. The sensitivity of the method was adequate for the determination of the drugs at toxic concentrations and at therapeutic levels. It is a simple, rapid, and reproducible method for the simultaneous determination of propranolol or metoprolol and some butyrophenones. The minimum detectable concentration was 50 ng/mL with a flame ionization detector and 10 ng/mL with a mass spectrometer detector. PMID- 8429499 TI - New degradation product of spiradoline mesylate in aqueous solution: formation of an imidazolidine ring. AB - A new degradation product of spiradoline mesylate was recognized in aqueous solution stored at 80 degrees C for 10 days. The structure of the degradation product was elucidated with spectroscopic data and a synthetic method as (1RS,3RS,8RS,10RS)-2-methyl-2,7-diazatricyclo - [6.4.0.0(3,7)]dodecane-10-spiro 2'-tetrahydrofurane by several two-dimensional NMR spectra. The fused imidazolidine ring system was synthesized from (5RS,7RS,8RS)-N-methyl-8-(7 pyrrolidino-1-oxa spiro-[4.5] decane)amine reacted with mercuric acetate. Because the degradation was promoted by bubbling air or oxygen gas into the solution or by adding alpha,alpha'-azobisisobutylonitrile for radical initiation, and the reaction was suppressed by bubbling nitrogen gas or deaerating, the reaction mechanism of the degradation was considered to be oxidation caused by an oxygen radical. PMID- 8429500 TI - Degradation kinetics of metronidazole in solution. AB - The degradation kinetics of metronidazole in aqueous solutions of pH 3.1 to 9.9 under accelerated storage conditions were studied. The stability of metronidazole in solutions containing propylene glycol or polyethylene glycol 400 was also investigated. The reaction order for metronidazole in these aqueous and solvent systems followed pseudo-first-order degradation kinetics. The degradation rate of metronidazole was invariant under various total buffer concentrations at each specific pH within the investigated pH range. These results indicate that no general acid/base catalysis imposed by acetate, phosphate, and borate buffer species is responsible for the degradation of metronidazole. The catalytic rate constants for hydrogen ion, water, and hydroxyl ion for the degradation of metronidazole were 6.11 x 10(-5) M/s, 3.54 x 10(-8) L/s, and 4.10 x 10(-3) M/s, respectively. The pH-rate profile shows a pH-independent region of pH 3.9-6.6. Maximum stability of metronidazole was at pH 5.6 under zero total buffer species conditions. The ionic strength effect on metronidazole degradation in acetate and phosphate buffers followed the modified Debye-Huckel equation well. The Arrhenius plot showing the temperature dependence of metronidazole degradation indicates estimates of activation energy of 15.35 kcal/mol and a half-life of 963 h at room temperature in 0.1 M pH 3.1 acetate buffer solution (ionic strength = 0.5). Irradiation with UV light (254 nm) of the metronidazole solutions (pH 3.1 acetate buffer) accelerated degradation in comparison with light-protected samples. Incorporation of propylene glycol into the metronidazole solution at pH 3.1 increased stability; however, an adverse effect on the stability of metronidazole was seen when polyethylene glycol 400 solvent system was used. PMID- 8429501 TI - Zatebradine: pharmacokinetics of a novel heart-rate-lowering agent after intravenous infusion and oral administration to healthy subjects. AB - Zatebradine (1; UL-FS 49 CL; 1,3,4,5-[tetrahydro-7,8-dimethoxy-3-[3-[ [2-(3,4 dimethoxyphenyl)ethyl]methylamino]propyl]-2H-3-benzazepin- 2-on- hydrochloride) is the proposed INN name for a nonchiral, novel, specific heart-rate-lowering drug that is suitable for the treatment of stable angina pectoris. The pharmacokinetics of 1 and of total radioactivity in healthy volunteers (n = 6) were determined after intravenous infusion and oral administration of an experimental drug solution containing 7.5 mg (1.85 MBq) of 14C-labeled drug. Concentrations in plasma and urine were measured by a specific, sensitive, reversed-phase automated high-performance liquid chromatography system with fluorimetric detection at 285 nm Ex, 315 nm Em and by liquid scintillation counting. Recovery of total radioactivity was 89.8 +/- 2.3% (infusion) and 92.2 +/- 3.0% (oral). Renal elimination of total radioactivity was 62.5 +/- 2.0% (infusion) and 48.8 +/- 3.1% (oral). After intravenous infusion and oral administration, 27.3 +/- 2.4 and 43.4 +/- 3.9%, respectively, of the administered dose was eliminated in the feces. Absorption, based on renal excretion of total radioactivity following oral and intravenous dosing, was 79.2 +/- 15.3% (mean +/- standard deviation). Unchanged parent drug contributed 28.4 +/- 5.8% (infusion) and 12.4 +/- 3.7% (oral) of the dose to renal excretion. Zero to three minutes after cessation of the 20-min infusion, the maximum concentration of parent drug in plasma was 161.9 +/- 70.9 ng/mL. After oral administration, a mean peak concentration in plasma of 24.3 +/- 6.7 ng/mL (0.5-3 h postadministration) was reached. Data regarding levels of 1 in plasma could be adequately fitted with an open, two-compartment model.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429502 TI - Abutment selection for anterior single tooth replacement. A clinical report. PMID- 8429503 TI - Use of existing pontics to maintain esthetics with a transition prosthesis: a clinical report. PMID- 8429504 TI - Amalgam restorations: a new approach. AB - A conservative approach to amalgam restorations is presented for using a chemomechanical method for softening and removing decay and using a dental adhesive resin for bonding amalgam to tooth structure, reducing the need for mechanical retention and preserving sound tooth structure. PMID- 8429505 TI - Intraoral galvanic corrosion: literature review and case report. AB - This article reviewed the dental history of a patient with symptomatic electro chemical reactions after the occlusal relationship of an existing complete gold crown and silver amalgam restoration was changed. A literature review of oral galvanism is presented with diagnostic techniques and treatment options. PMID- 8429506 TI - Resin-bonded fixed partial denture technique: results of a medium-term clinical follow-up investigation. AB - Results of a clinical investigation with cast metal, electrolytically etched, resin-bonded fixed partial dentures (FPDs) are reported. These FPDs were inserted under standardized conditions. The FPDs had been in place and functioning for an average time of 21.4 months. Sixty-five patients with a total of 82 resin-bonded FPDs were followed up. There was a retentive failure in only one FPD. All patients were satisfied from both functional and esthetic points of view. The results of this study indicate that the concept of treatment with resin-bonded FPDs should be considered as an alternative to conventional treatment. PMID- 8429507 TI - Retentive properties of a threaded split post with attachment sleeves cemented with various luting agents. AB - This study determined the retention, in pounds, of Nos. 1 through 3 stainless steel attachment sleeves for the threaded split post system cemented with various cements. Post heads and matching sleeves were also sandblasted and compared with attachments not sandblasted. Four luting agents were selected, zinc oxyphosphate cement (ZOP), and three brands of composite resin cement (CRC). The experiment was divided into two parts; in part 1, samples were not treated, and in part 2, samples were sandblasted. Retentive values were recorded with a universal testing machine. A 2 x 3 x 4 factorial three-way analysis of variance was used to compute the effects sandblasting, sleeve and post type, and the luting agent. No statistical differences were recorded for sleeves not sandblasted, Nos. 1 and 2, regardless of the luting agent. The nonsandblasted No. 3 sleeve cemented with Panavia CRC had significantly greater retention than All-Bond CRC or ZOP. There were no statistical differences in retention of sandblasted sleeves for the No. 1 sleeve despite the type of luting agent. For sandblasted sleeves Nos. 2 and 3, ZOP and Panavia CRC recorded significantly greater retention than All-Bond CRC. However, there was no difference between sandblasting and not sandblasting for the No. 1 sleeve. Mean loads for Nos. 2 and 3 sleeves were substantially greater for sandblasted compared with attachments not sandblasted. PMID- 8429508 TI - Saucer-shaped cavity preparation for composite resin restorations in class II carious lesions: three-year results. AB - This study determined the feasibility of saucer-shaped cavity preparations for composite resins in class II lesions. Saucer-shaped class II cavity preparations were restored with composite resin and subjected to clinical, radiographic, and replica cast evaluation (39 restorations) after 6, 18, and 36 months. The results indicated that the retention, resistance form, and wear resistance of the class II saucer-shaped cavity preparations were satisfactory after 3 years. PMID- 8429509 TI - Alternatives to posterior complete crowns: integrating foundations with cuspal protection. AB - A common approach for restoring compromised posterior teeth is to reconstruct the missing anatomic crown with a dental restorative material and prepare the remaining tooth structure for an artificial veneer. Because this procedure is also compounded by designing subgingival finish lines for tooth preparations, a conservative approach for the fabrication of one-piece castings is suggested that incorporates both foundations and cuspal protection, with finish lines prepared above the gingival crest. Patient selection, advantages, disadvantages, and armamentarium are discussed. PMID- 8429510 TI - Preliminary study of electromyographic characteristics for distinguishing centric relation and protrusion in edentulous patients. AB - The mean voltages of the masseter and anterior temporal muscles of 30 Chinese edentulous patients were recorded in centric relation and protrusive positions (from 1 mm to 4 mm anterior to centric relation) under two given occlusal force conditions. It was found that the mean ratio of individual voltages of the anterior temporal muscles to the masseter muscles was equal to or greater than 1.2 in centric relation, but the ratio was less than 1.2 in protrusive positions in most of the subjects. The results demonstrated that the ratio of voltages of the anterior temporal muscles to masseter muscles may be a reliable parameter for distinguishing centric relation from protrusion in Chinese edentulous patients. PMID- 8429511 TI - Statistical analyses on the success potential of osseointegrated implants: a retrospective single-dimension statistical analysis. AB - Osseointegrated implants are a form of therapy that is finding increasing application in dental treatment. Long-term success can be ensured if the results of recall examinations are systematically documented. Life table analyses are being used to a growing extent to ensure objective assessment of the success of osseointegrated implants. With the help of the statistical method designed by Kaplan-Meier and Cutler-Ederer in 1958, the survival time of the implant can be calculated and the relationship between various covariables and the service time of the implants can be determined. This study calculates the success potential of 683 osseointegrated implants (IMZ and Branemark) using the customary input-output statistics and life table analyses to explain the discrepancy between the two statistical methods, based on the results obtained. PMID- 8429512 TI - An appliance based approach to the management of palatopharyngeal incompetency: a clinical pilot project. AB - The palatal lift appliance has been described in the dental literature for more than 30 years, yet its efficacy remains controversial in the management of palatopharyngeal incompetency during speech. A pilot project was undertaken to develop a clinical research protocol. In the pilot project, 32 patients had palatopharyngeal incompetency managed with a palatal lift appliance. The results of the treatment were that 21 of the 32 patients experienced reduction programs but, of these, seven still required surgery. Before firm conclusions on the use of the palatal lift appliance can be made, objective, measurable assessment of speech through well-designed clinical protocols is required. PMID- 8429513 TI - Audiometric evaluation of prosthetic ears: a preliminary report. AB - A pilot study was conducted to measure the acoustic change provided by a prosthetic ear and to determine whether this change is functionally significant. Four subjects with acquired loss of the ear were tested. Objective data confirmed that the prosthesis provided acoustic gain. This gain may be significant only for individuals with a preexisting hearing loss. PMID- 8429514 TI - Effect of lateral cranial base surgery on temporomandibular joint function. AB - Temporomandibular joint (TMJ) function in patients who experienced lateral cranial base tumor surgery was evaluated by a comparison of recordings of condylar movement in right and left parasagittal planes during opening and protrusive movements. The characteristic tracing and measurements of mandibular movements at the condyle of six patients after surgery were compared with a nonsurgical asymptomatic control group consisting of 16 subjects. Of the six surgical patients, three patients had the condyles disarticulated and rearticulated, and three patients had condyles surgically resected. The data gathered through computerized axiographic tracings of horizontal condylar inclinations at different intervals showed significant interaction effects among right and left sides within the groups (p 0.001). Medical histories and clinical examinations were used to diagnose or rule out the presence of TMJ disorders. Magnetic resonance imaging (MRI) and three-dimensional computerized tomograms (CT) imaging of the TMJs of these patients were investigated. MRI and CT scans illustrated the static detail and morphology of the TMJs. Computerized axiography revealed the dynamic detail and function of the TMJs in the various mandibular movements. The study of condylar pathways during opening and protrusive movements showed variations of mandibular deviation among patients having cranial base surgery. Increased mandibular deviation was found in patients who experienced resected condyles. PMID- 8429515 TI - Axiographic tracings of temporomandibular joint movements. AB - Three-dimensional condylar movements of 49 symptomatic and asymptomatic volunteers were recorded with a hinge axis tracing system axiograph during maximal opening, protrusion, and mediotrusion. The tracings displayed in sagittal and frontal planes were measured to evaluate biomechanics of the temporomandibular joint. The only differences in condylar tracings between symptomatic and asymptomatic groups were in the right joint, recorded in the sagittal plane during maximal opening, and the Bennett angle. The symptomatic group had a significantly longer condylar path and a smaller Bennett angle compared with the asymptomatic group. The results were interpreted as indications of adaptive morphologic instead of pathologic changes. The alterations in condylar tracings as an indicator of joint pathology should be considered cautiously. PMID- 8429516 TI - Deep thermometry of temporomandibular joint and masticatory muscle regions. AB - A study was designed to measure noninvasively the deep temperature of the temporomandibular joint (TMJ) region and corresponding regions of the masticatory muscles at rest. With a transcutaneous probe, the deep thermometry of the right and left anterior (Ta) and the posterior portion (Tp) of the temporal muscles, the mid-portion of the superficial belly of the masseter muscles (Mm), and the TMJ regions were measured. In 20 normal male subjects, the deep temperature of the Ta region (mean 36.342 degrees C), the Tp region (mean 36.345 degrees C), and the TMJ region (mean 36.06 degrees C) was higher than that of the Mm region (mean 35.897 degrees C) at rest. In addition, no differences in the deep temperature were observed between the right and left Ta, Tp, Mm, and TMJ regions at rest. All of the normal subjects showed differences between the right and left TMJ region of less than 0.3 degrees C. In 10 patients with craniomandibular disorders, however, eight patients showed differences of more than 0.3 degrees C between the asymptomatic and asymptomatic TMJ region. Because of high sensitivity and specificity, the deep thermometry measurements can provide useful non-invasive information. PMID- 8429517 TI - Protection of recent extraction sites when fabricating immediate acrylic interim fixed restorations. AB - Protection of all soft tissues is an early and integral component of the total treatment plan involving any dental restoration. A procedure is presented to provide protection of recent extraction sites by using adhesive Burlew foil when making an immediate fixed restoration. The foil provides protection, at the extraction site, from both mechanical and chemical irritation. PMID- 8429518 TI - A method to remold worn acrylic resin posterior denture teeth and restore lost vertical dimension of occlusion. AB - A method is described for remolding and correctly restoring abraded posterior denture teeth. By adding a layer of paraffin wax for jaw registration, the dentist can repeat the registration procedure until the proper jaw relation is established. This wax occlusal record is used as a guide during the remolding, and is incrementally replaced by autopolymerizing resin. PMID- 8429519 TI - Rubber dam isolation of fixed partial denture. PMID- 8429520 TI - A modified post puller. PMID- 8429521 TI - Fetal mononuclear cells show a comparable capacity with maternal mononuclear cells to produce IL-8 in response to lipopolysaccharide in chorioamnionitis. AB - IL-8 is a chemotactic and activating cytokine for neutrophils which eliminate invading bacteria by releasing bactericidal metabolites. Cord blood mononuclear cells (CBMCs) obtained from neonates born to mothers with chorioamnionitis actively produced a significantly higher amount of IL-8 than those of neonates without chorioamnionitis, suggesting that the mononuclear cells of fetuses with chorioamnionitis had been activated in utero. As lipopolysaccharide (LPS) can often be detected in the uteroplacental space in chorioamnionitis, the LPS mediated activation mechanism of neonatal mononuclear cells was analyzed in vitro to produce IL-8. Neonatal mononuclear cells stimulated with LPS increased IL-8 production in a time- and dose-dependent manner. The ability of term or preterm neonatal mononuclear cells to produce IL-8 was comparable with that of adult (maternal) mononuclear cells, suggesting functional maturity of the neonatal or fetal mononuclear cells to produce IL-8. However, IL-8 production by neonatal CBMCs was down-regulated by dexamethasone, a glucocorticoid which is clinically administered to mothers to promote fetal lung maturity in preterm delivery. Our present study revealed a regulatory mechanism of fetal IL-8 production, suggesting that functionally mature fetal mononuclear cells produce IL-8 in response to LPS in chorioamnionitis and activate the fetal defense mechanism against infection. PMID- 8429522 TI - Interleukin-6 in human milk. AB - Interleukin-6 (IL-6) in human milk collected during the first 2 days of lactation was investigated by a competitive radioimmunoassay (RIA) and column chromatography. The concentrations of IL-6 in the aqueous phase of fresh human milk were 151 +/- 89 pg/ml. The concentrations of IL-6 in milk increased after storage at 4 degrees C and decreased after storage at -20 degrees C (P < 0.01). Column chromatography revealed two molecular weight peaks of IL-6 in human milk, the first > or = 100 kDa and the second between 25 and 30 kDa. The 25-30-kDa peak corresponded to known isoforms of human IL-6 and to the elution pattern for 125I labeled recombinant human IL-6, whereas the higher molecular weight peak may be in keeping with a bound or compartmentalized form of that cytokine. The precise molecular forms of this protein, the compartmentalization of or binding proteins for this cytokine and the in vivo effects of IL-6 in human milk upon the mammary gland or the recipient infant remain to be explored. PMID- 8429523 TI - An immunohistochemical study of leucocytes in human endometrium, first and third trimester basal decidua. AB - An immunohistochemical quantitative study of leucocyte subpopulations on fresh human endometrium and on biopsy specimens of first and third trimester basal decidua in normal (uncomplicated) pregnancies was performed. The most prominent population in endometrial and decidual stroma of basal decidua are macrophages. B cells as well as gamma/delta T cell receptor positive cells were found occasionally, scattered throughout the endometrial/decidual stroma. CD3+ cells were present in a relatively small number in the endometrium as well as in the first trimester basal decidua, but their number was elevated (doubled) in the third trimester of pregnancy. CD2+ cells showed a slight increase in first trimester basal decidua when compared with both endometrium and third trimester basal decidua. Cells with positive NKH-1 marker (CD56+) showed a significant increase in the first trimester, while in the third trimester their number diminished drastically. CD56:CD3 cell ratio increased to more than five times in first trimester basal decidua, while in the third trimester basal decidua decreased drastically. The mentioned increase of CD56+ cells in the first trimester and that of CD3+ cells at term suggests that these cells could have some specific function(s). However, it still has to be established whether the described quantitative changes of decidual leucocytes in basal decidua during pregnancy are of any importance for the mechanism(s) for the fetal allograft protection. PMID- 8429524 TI - Effect of IFN-gamma and IFN-alpha on killing of human trophoblast by decidual LAK cells. AB - Human decidual large granular lymphocytes are capable of killing both normal trophoblast and choriocarcinoma cells after stimulation with recombinant interleukin-2 (rIL-2) in vitro. We now show that pre-treatment of normal trophoblast and JEG-3 choriocarcinoma cells with interferon-gamma (IFN-gamma) protects these target cells from killing by decidual effectors, although pre treatment with interferon-alpha (IFN-alpha) had no such effect. In contrast, JAR choriocarcinoma cells were readily killed before and after exposure to both IFN gamma and IFN-alpha. This protective effect might be due to upregulation of non classical trophoblast HLA Class I molecules, as IFN-gamma but not IFN-alpha enhances Class I surface expression in both normal trophoblast and JEG-3 choriocarcinoma cells. JAR cells, however, are constitutively and inductively negative for Class I expression. PMID- 8429525 TI - Secretion of tumor necrosis factor alpha by testicular macrophages. AB - While it has been shown that culture medium from testicular macrophages can influence testosterone production when added to Leydig cells, the identity of the factor(s) responsible for this activity remains unknown. Since tumor necrosis factor alpha (TNF alpha) has been shown to be capable of influencing testosterone production by Leydig cells, a series of studies was conducted to determine if testicular macrophages produce TNF alpha. It was found that testicular macrophages from adult rats produce a factor that is capable of lysing L929 cells, which are used as a traditional bioassay for TNF alpha. The TNF alpha activity in the macrophage-conditioned medium could be neutralized by the addition of anti-murine TNF alpha but not with the addition of preimmune IgG. While lipopolysaccharide (LPS) slightly increased the release of TNF alpha, neither follicle-stimulating hormone (FSH) nor testosterone had a similar effect. It was not determined if the isolation procedure had artificially 'activated' the macrophages. Medium from cultured Sertoli cells, Leydig cells and peritubular cells did not contain TNF alpha activity. These studies are consistent with the hypothesis that the paracrine interaction between testicular macrophages and Leydig cells is mediated in part by TNF alpha. PMID- 8429526 TI - Unusual expression pattern of the MHC class I Q5k gene in the AKR mouse: possible role in embryo development. AB - Analysis of the tissue-specific expression of the Q5k gene in the AKR mouse reveals an unusual expression pattern. The Q5k mRNA is present in embryos from day 12, but expression is switched off in most tissues except thymus and testis shortly after birth. Late in pregnancy the gene is again transcribed in females. Analysis at the epitope level, with a Qa-2 specific monoclonal antibody revealed that in most cases the Q5k product is confined to the cytoplasm. These results suggest that Q5k has a most unusual tissue distribution and timing of expression among all the H-2 class I and Q genes so far described. PMID- 8429527 TI - Production and determination of specificity of monoclonal IgG anti-mouse blastocyst antibodies. AB - Five monoclonal anti-mouse-blastocyst IgG antibodies were raised by intrasplenic immunization of three mice with adhesive-stage mouse blastocysts. Each mouse received a total of 60-70 blastocysts which were either nitrocellulose immobilized or living but irradiated. Tests for pre-implantation stage specificity showed that the antibodies differed in specificity. None were specific for surface epitopes. One antibody recognized epitopes only on blastocysts. Other antibodies were able to discriminate between unfertilized and fertilized oocytes, uncompacted and compacted morulae, or delayed and adhesive blastocysts. By applying reduced SDS-PAGE and Western blotting to blastocysts the blastocyst-specific antibody was seen to be bound to a peptide of M(r) 34. PMID- 8429528 TI - Cervical ripening: advances in preinduction of labor. Proceedings from a roundtable discussion. PMID- 8429529 TI - Spontaneous and induced cervical ripening. Natural dilation and effacement process and current cervical ripening techniques. AB - During gestation, the cervix forms a tight sphincter to ensure the integrity of the pregnancy. Toward the end of the pregnancy, hormone-mediated biochemical changes cause the cervix to ripen and become softer and more pliable to allow passage of the fetus. Failure of the cervix to ripen may result in delayed onset of labor and a prolonged and complicated course, especially if labor is artificially induced. Attempts to induce cervical ripening have involved the use of mechanical methods, estrogen and estrogen precursors, relaxin and prostaglandins. The ideal ripening agent is simple and noninvasive, effective within 24 hours, does not compromise mother or fetus and does not stimulate labor (during the ripening process). PMID- 8429530 TI - Therapeutic considerations for preinduction cervical ripening with intracervical prostaglandin E2 gel. AB - A commercially prepared prostaglandin E2 (PGE2) gel for cervical ripening has been approved by the U.S. Food and Drug Administration (FDA). In circumstances when labor must be induced before the cervix has undergone spontaneous softening and effacement, this product effectively prepares the cervix for labor. Compared with extemporaneous preparations for vaginal administration, this product offers stability, uniform dosage and minimal side effects. Intracervical PGE2 gel offers significant clinical benefits, but also requires consideration of important clinical issues related to the selection of patients, application of the gel and subsequent monitoring of the patient and fetus. PMID- 8429531 TI - Endocervical prostaglandin E2 gel for preinduction cervical ripening. Clinical trial results. AB - Randomized, clinical trial results of a standardized, stable prostaglandin E2 (PGE2) preparation (0.5-mg PGE2 in a 2.5-mL gel base) applied endocervically as a single dose 12 hours prior to oxytocin induction were studied. The trials, conducted in the United States and Canada, included 538 women; 277 treated with PGE2 gel and 261 controls, with initial Bishop scores of < or = 4. Compared to controls, PGE2 recipients had significant improvement in cervical Bishop scores (P < .01) and a high rate of labor (47.4% vs. 9.6%, P < .001) during the 12-hour ripening period. In reporting centers, delivery during this same interval occurred without use of oxytocin in PGE2 patients (25.4%) vs. controls (4.9%). PGE2 reduced induction to delivery intervals and the duration of oxytocin administration. Although the composite cesarean section rates were lower in PGE2 recipients than in controls (28.5% vs. 32.9%), this difference did not achieve statistical significance. Maternal and fetal outcome and the incidence of complications were similar in the two groups. These studies confirm the safety and efficacy of this standardized PGE2 preparation as a means of cervical ripening in parturients with highly unfavorable cervixes. PMID- 8429532 TI - Prostaglandin E2 preparations for preinduction cervical ripening. Pharmacy considerations. AB - The use of locally applied prostaglandin E2 (PGE2) has become a common intervention in the management of the unripened cervix in term pregnancy. The extemporaneously prepared, vaginally administered prostaglandin E2 gels are most often used clinically in the United States. Large-scale clinical trials have not been conducted with these products. The extemporaneously prepared gels are not standardized with regard to potency, purity or stability; the assessment of efficacy and safety is based on anecdotal clinical reports. PGE2 and its metabolites are eliminated rapidly from the circulation, thus plasma levels are not useful in assessing clinical efficacy. Pharmacokinetic evaluations are based on plasma levels of the stable bicyclo-metabolite of PGE2. Plasma levels resulting from exogenous administration of PGE2 cannot be distinguished from plasma levels caused by endogenous production. Available evidence supports local application of low doses of PGE2 for cervical ripening prior to labor induction. Local application may minimize undesirable systemic effects and uterine hypertonus. A cost/benefit evaluation of a commercially prepared intracervical PGE2 gel must consider several factors in addition to acquisition cost including avoidance of other more costly procedures, such as cesarean section, instrumental delivery, anesthesia, transfusions and monitoring of prolonged labor and fetal outcome, liability issues and pharmacy preparation costs. PMID- 8429533 TI - Prostaglandins in preinduction cervical ripening. Meta-analysis of worldwide clinical experience. AB - The state of the cervix is an important predictor of success in the induction of labor. A firm and rigid (unripe) cervix increases the likelihood of a failed induction or a prolonged, exhausting labor. Administration of prostaglandins has shown good effects when used for cervical ripening. A meta-analysis of results of studies comparing prostaglandin therapy to placebo or no treatment and comparing different prostaglandin preparations and routes of administration is presented. Currently, prostaglandin E2 appears preferable to prostaglandin F2 alpha based on effectiveness at lower doses, thus minimizing side effects resulting from influences on other body systems. Further research is required to determine the optimal route of prostaglandin administration, although the oral route appears to be unsuitable. Prostaglandins administered vaginally or endocervically appear to offer the best balance of effectiveness and safety. PMID- 8429534 TI - In-vitro and in-vivo characterisation of resistance to colonisation with Clostridium difficile. AB - In hamsters, resistance to colonisation by Clostridium difficile appears to be mediated by micro-organisms that are present in the gut in relatively low concentrations. Small amounts of normal caecal contents inhibited the growth of C. difficile when added to cultures in vitro or given to animals which had been treated with clindamycin. Filtrates of caecal contents, frozen and thawed contents and contents diluted to 0.1% wet weight lost their inhibitory properties. However, caecal contents retained their protective capacity after culture for 7 days in vitro. Antibiotic treatment altered resistance to colonisation by only a few species of clostridia. Faeces of animals treated with ampicillin but not clindamycin recovered colonisation resistance after incubation at 37 degrees C in vitro. Since human faeces could also restore colonisation resistance to hamsters, the hamster model may be useful for the study of resistance to colonisation by C. difficile in man. PMID- 8429535 TI - Characterisation of Clostridium difficile strains by polymerase chain reaction with toxin A- and B-specific primers. AB - A total of 218 Clostridium difficile strains was examined for production of toxin A by ELISA, production of toxin B by a cytotoxin assay and the presence of toxin A and B gene-associated sequences by the polymerase chain reaction (PCR). After saturation amplification with toxin B-specific primers, the characteristic amplification product (591 bp) was detected in all 184 toxigenic strains examined. PCR with toxin A-specific primers gave positive results with all but one of the toxigenic strains. By contrast, PCR with toxin A- and toxin B-specific primers yielded negative results with all 34 non-toxigenic strains tested. This suggests that PCR detection of either the toxin A or B gene is a good indication of toxin production. PCR did not require DNA extraction or hybridisation and was convenient, sensitive and rapid. Toxigenic C. difficile could be detected in mixed cultures, suggesting a role for PCR in the identification of toxigenic C. difficile in primary culture. PMID- 8429536 TI - Stability of cefdinir (C1-983, FK482) to extended-spectrum plasmid-mediated beta lactamases. AB - Fourteen plasmid-encoded extended-spectrum beta-lactamases were purified from Escherichia coli transconjugants of original clinical isolates. The Vmax, Km and Vmax/Km were each determined for ampicillin, carbenicillin, cephaloridine, cephalexin, cefuroxime, cefixime, cefdinir, ceftazidime and cefotaxime as substrates with eight of these enzymes and with the narrow-spectrum beta lactamase, TEM-1. The relative rates of hydrolysis of ampicillin, cephaloridine, cephalexin, cefuroxime, cefixime, cefdinir, ceftazidime and cefotaxime were also determined for the remaining six enzymes. Cefdinir had Vmax/Km or relative rates of hydrolysis values either equal to or lower than ampicillin, cephaloridine, cephalexin and cefotaxime for all the enzymes tested. Overall, cefdinir was more stable to the 15 beta-lactamases tested than either cefuroxime or cefixime; however, ceftazidime was more stable than cefdinir to hydrolysis by eight of the enzymes tested. PMID- 8429537 TI - Genetic characterisation of isolates of Listeria monocytogenes from man, animals and food. AB - Multilocus enzyme electrophoresis was used to assess genetic relationships between 95 isolates of Listeria monocytogenes, most of which were isolated in Australia and New Zealand from man, animals and food. The isolates were separable into two major genetic divisions; the majority of those from human patients and animals were in division I, and the majority from those foods that were not specifically associated with human listeriosis were in division II. Isolates in division I were virulent, whereas many isolates from food were probably less virulent and did not pose a large threat to human health. However, isolates from certain foods, particularly pate, were indistinguishable from those causing disease in man, and the consumption of these products represented a clear risk factor for infection. Isolates from infected human patients in Australia and New Zealand belonged to the same clone of serotype 4b that has been responsible for major epidemics in the northern hemisphere. However, a separate clone of serotype 1/2b strains, present in both Australia and New Zealand, was responsible for two major outbreaks that occurred in Western Australia in 1978-80 and 1990-91. PMID- 8429538 TI - Evolutionary origin and radiation of the avian-adapted non-motile salmonellae. AB - Multilocus enzyme electrophoresis was employed to estimate chromosomal genotypic diversity and relationships among 131 isolates of the non-motile Salmonella biotypes Gallinarum and Pullorum (serotype 1, 9, 12:-:-) that cause fowl typhoid and pullorum disease, respectively. Thirteen electrophoretic types (ETs), marking clones, were distinguished, and construction of a neighbour-joining phylogenetic tree revealed three lineages: one consisted of five ETs of Gallinarum, a second included seven ETs of Pullorum, and a third was represented by a single ET (Ga/Pu 1) that is intermediate between those of the other two lineages in both multilocus enzyme genotype and biochemical properties. Enzyme genotype analysis and comparative nucleotide sequencing of the phase 1 flagellin gene (fliC), the hook-associated protein 1 gene (flgK), and the 6-phosphogluconate dehydrogenase gene (gnd) identified serotype Enteritidis (1, 9, 12:g, m:-) as a close relative of the non-motile salmonellae. In most strains of biotype Gallinarum, the fliC gene is complete, intact and identical in sequence to that of Enteritidis, but isolates of three ETs had a stop codon at position 495. The fliC sequences of the ETs of Pullorum differed from that of Enteritidis in having non-synonymous changes in either two or three codons and a synonymous change in one codon. The sharing of distinctive alleles at three metabolic enzyme loci and a stop codon in flgK indicates that the non-motile salmonellae are monophyletic and that their most recent common ancestor was non-motile. Since diverging from that ancestor, the Pullorum lineage has evolved more rapidly than the Gallinarum and Ga/Pu 1 lineages. PMID- 8429539 TI - Identification of Bordetella pertussis in nasopharyngeal swabs by PCR amplification of a region of the adenylate cyclase gene. AB - The polymerase chain reaction (PCR) was used to amplify a 522-bp region of the adenylate cyclase toxin (cyaA) gene of Bordetella pertussis. As few as 100 cfu from a suspension of B. pertussis could be detected by this procedure when the amplified PCR product was detected by ethidium bromide staining of agarose gels. However, simulated clinical specimens, prepared from swabs impregnated with known numbers of B. pertussis cells, only yielded a positive reaction with > or = 10(4) cfu. Hybridisation of a Southern blot of the PCR products from the swab samples with a cya-specific probe gave a positive reaction with as few as 8 cfu, but the hybridisation signal was uniformly weak with fewer than 10(4) cfu. Nevertheless, three of 13 nasopharyngeal swabs, taken from suspected clinically defined cases of whooping cough and stored frozen for up to 18 months, gave a positive PCR reaction. PMID- 8429540 TI - Campylobacter jejuni adapts to aerobic metabolism in the environment. AB - Campylobacter jejuni, when left on blood agar for prolonged periods, was found to survive better in air than under micro-aerobic conditions. After a period of 2-3 days in air, all strains of C. jejuni examined grew freely in air on subculture, and could be further subcultured apparently indefinitely in air. This adaptation to aerobic metabolism was accompanied by a change in colony morphology and some changes in outer-membrane protein patterns, but no change in serotyping reactions. The ability to colonise mice was unaltered as was the helical morphology of growing cells. The important survival phase of C. jejuni, when outside the animal gut, involves not only a change to coccal morphology but also fundamental changes in the metabolism of the organism. These changes are likely to be relevant to techniques required for culturing C. jejuni from foods and environmental sources. PMID- 8429541 TI - Phagocytosis of bacterial strains isolated from acute dentoalveolar abscess. AB - The phagocytosis by human polymorphonuclear leucocytes of 37 bacterial strains identified as Streptococcus milleri (10 strains), strictly anaerobic gram positive cocci (10) Prevotella intermedia (6), Pr. oralis (5) and Fusobacterium nucleatum (6) was investigated in vitro. The ingestion of S. milleri and strictly anaerobic gram-positive cocci was significantly greater (p < 0.001) than that of strains of Prevotella spp. and F. nucleatum. The degree of uptake of capsulate and non-capsulate strains did not differ. PMID- 8429542 TI - Meningococcal vaccines. PMID- 8429543 TI - Vaccine efficacies of elastase, exotoxin A, and outer-membrane protein F in preventing chronic pulmonary infection by Pseudomonas aeruginosa in a rat model. AB - The protective efficacies of eight vaccine preparations consisting of Pseudomonas aeruginosa outer-membrane protein F, elastase and exotoxin A toxoid, administered either individually or in various combinations, were determined in a rat model of chronic pulmonary infection. Rats were immunised intramuscularly at 2-week intervals (days 0, 14 and 28). On day 42, blood was collected and antisera were obtained from each vaccine group for use in an enzyme-linked immunosorbent assay which determined the titre of IgG antibodies elicited by each vaccine. Also on day 42, rats were challenged by intratracheal inoculation of a clinical isolate of P. aeruginosa encased within agar beads. On day 49, the animals were killed and the lungs were examined macroscopically for the presence of lesions and fixed for histological examination. When compared with control rats immunised with bovine serum albumin, rats immunised with protein F alone as a vaccine received significant protection against the development of severe pulmonary lesions. Elastase used alone as a vaccine provided some protection against severe lung lesions and reduced the incidence of microscopic peribronchial inflammation. However, the combination of protein F plus elastase as a vaccine did not afford protection from severe lesions, and there was an increased incidence of necrotising granulomas in the lungs from this vaccine group. Protection against lung lesions from the three-component vaccine consisting of protein F, elastase and exotoxin A toxoid was similar to that provided by the protein F vaccine. Neither macroscopic nor histological evidence showed any enhancement of protective efficacy for the three-component vaccine over that of the protein F vaccine.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429544 TI - Detection of enterovirulent Escherichia coli associated with diarrhoea in Seville, southern Spain, with non-radioactive DNA probes. AB - To assess the role of diarrhoeagenic Escherichia coli in Southern Spain, faecal samples from 135 patients with diarrhoea and 40 healthy subjects from Seville, Andalusia, were investigated. In this prospective study, enterovirulent E. coli were identified by hybridisation with five non-radioactive DNA probes specific for enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC) and verocytotoxin-producing E. coli (VTEC). Probe-positive strains were isolated from four patients (3%) with diarrhoea and from none of the healthy controls. Two patients harboured ETEC and two patients had EPEC probe-positive strains in their faeces. No VTEC were isolated during this study. Salmonella spp. were the most frequently identified enteric pathogens, accounting for 10% of the cases, followed by Campylobacter jejuni (3%) and diarrhoeagenic E. coli (3%). This study indicates that enterovirulent E. coli play a modest role in the aetiology of diarrhoea among the indigenous population of Southern Spain. PMID- 8429545 TI - Serum and tissue protein binding and cell surface properties of Staphylococcus lugdunensis. AB - Eleven strains of Staphylococcus lugdunensis from different clinical sources were investigated for their ability to bind 125I-labelled collagen (Cn) type I and IV, fibronectin (Fn), vitronectin (Vn), laminin (Lm), fibrinogen (Fg), thrombospondin, plasminogen (glu- and lys-form) and human IgG. All the strains bound these proteins, although a higher degree of binding was obtained for Cn types I and IV and IgG with mean values of 36%, 32% and 26% binding, respectively. In tests with proteins immobilised on latex beads in a particle agglutination assay, eight of the 11 strains bound Cn type I and seven bound Fg, whereas no strain bound immobilised IgG. Binding to immobilised Cn-I, Fg, Lm and Vn was abolished when the bacterial cells were treated with proteases or heat, indicating cell-surface receptors with protein characteristics. Cell-surface extracts of S. lugdunensis 2342 were able to totally inhibit binding of the homologous strain and S. aureus Cowan 1 to latex-immobilised proteins Cn-I, Lm, Vn, Fn and Fg. The binding of 125I-labelled Cn IV by S. lugdunensis 2342, was heat sensitive, whereas the binding to S. aureus Cowan 1 was heat resistant. The strains gave negative results in tests for the presence of protein A with a S. aureus protein A gene probe and with sensitised red blood cells. No production of heat-stable nuclease (TNase) could be detected by monoclonal antibodies against TNase or by the polymerase chain reaction with an oligonucleotide sequence from S. aureus TNase as primer.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429546 TI - Calcium ions and the structure of muscle actin filament. An X-ray diffraction study. AB - In order to investigate the effects of Ca(2+)-binding to troponin on the conformation of the muscle thin filament (consisting of actin, tropomyosin and troponin) in the absence of actomyosin interaction, two series of X-ray diffraction experiments were undertaken. Firstly, the small angle X-ray scattering from filaments in solution indicate the tropomyosin strands are centred at about 3.5 nm from the filament axis and this distance is calcium independent. Secondly, X-ray fibre diffraction patterns from the filaments orientated in glass capillaries were studied. The X-ray intensity of the 2nd actin layer-line increased in a highly co-operative manner at a concentration of free calcium ions [Ca2+] of 10(-6.8) M, which is the range in which muscle contraction is physiologically regulated. However, this intensity increase accounted for some 30% of the total increase observed in diffraction patterns from muscle on activation, suggesting that the Ca(2+)-binding alters the state of the thin filament, which then undergoes further changes upon interaction with myosin. PMID- 8429547 TI - New approach for atomic force microscopy of membrane proteins. The imaging of cholera toxin. AB - We demonstrate that supported synthetic phospholipid bilayers, which are stabilized by lateral cross-linking in both leaflets, can be used for specimen preparation for atomic force microscopy of purified membrane proteins with high stability and excellent reproducibility under water or low-salt buffer. A bilayer containing 1,2-dipentacosa-10,12-diynoyl-phosphatidylcholine and 20 mol % ganglioside (GM1) was transferred onto the surface of mica from a Langmuir trough. Cholera toxin, both the B-subunit and the complete molecular randomly bound to the gangliosides, were imaged by atomic force microscopy in solution with a resolution of better than 2 nm. The pentameric structure of the B-subunit oligomers was well resolved. This result indicates that, with this preparation procedure, other membrane proteins may be studied at intermediate to high resolution under physiologically relevant conditions without the need for crystallization. PMID- 8429548 TI - X-rays destroy the lamellar structure of model membranes. AB - Lipid membranes undergo a dramatic lamellar-to-non-lamellar phase transformation upon being irradiated with X-rays. The structure change was evidenced by chemical breakdown of the lipid and by a change in the X-ray diffraction properties of the model membrane indicating complete disruption of lamellar stacking. Considering the importance of X-ray diffraction as a structure probe the problem of radiation damage to biological materials must be duly recognized. Because X-ray damage has been shown to be free radical-mediated these results suggest a means by which free radicals accumulating in cells during aging might compromise membrane integrity and contribute to cell death. PMID- 8429549 TI - Lrp, a global regulatory protein of Escherichia coli, binds co-operatively to multiple sites and activates transcription of ilvIH. AB - Lrp (Leucine-responsive regulatory protein) has recently been recognized as a major regulatory protein that controls the expression of many operons in Escherichia coli. Footprinting and gel retardation experiments with DNA from ilvIH, one of the operons controlled positively by Lrp, indicate that Lrp binds to six sites over a 200 base-pair region upstream from the promoter. Binding of Lrp to some of these sites is highly co-operative. We suggest a consensus sequence for Lrp binding based upon a comparison of six binding sites. An analysis of mutants indicates that five out of six binding sites are important for transcription activation and that two or three adjacent Lrp binding sites act synergistically in vivo. The observed synergistic effects in vivo may result from co-operative binding of Lrp to adjacent sites. We propose a model in which multiple binding sites contribute to the formation of a nucleoprotein complex, but only a particular proximal site positions Lrp properly so that it interacts with RNA polymerase. PMID- 8429550 TI - Cloning and characterization of a novel Bacillus thuringiensis cytolytic delta endotoxin. AB - A gene encoding a major entomocidal polypeptide from Bacillus thuringiensis subspecies kyushuensis delta-endotoxin crystals (CytB) was cloned into Escherichia coli and sequenced. The deduced amino acid sequence gave a predicted molecular mass of 29,236 Da and showed 39% identity and 70% similarity with the 27,371 Da CytA protein from Bacillus thuringiensis subspecies israelensis. The larger size of CytB compared to CytA appears to be due to additional sequence in CytB after the CytA C-terminus. Unlike CytA, CytB was freely expressed in Escherichia coli and formed cytoplasmic inclusions without the need for a "helper" protein. Electron microscopic observation of CytB inclusions revealed them to be generally amorphous, but examples possessing some lattice structure were seen. PMID- 8429551 TI - Multiple transitions to non-B-DNA structures occur in the distal regulatory region of the rat prolactin gene. AB - The developmentally regulated rat prolactin (rPRL) gene presents a promising model system toward understanding the biological role of non-B-DNA structural elements. Two predominantly alternating purine-pyrimidine (APP) (dA-dC)n.(dG-dT)n repeats of 58 and 178 base-pairs flank the (A + T)-rich distal regulatory region. We have characterized several transitions to non-B-DNA structures within this region in negatively supercoiled plasmids by utilizing high resolution chemical probing. Each repeat undergoes a full-length conversion to a novel left-handed helical structure via the stepwise nucleation and propagation of discrete "segments". These segments are delimited by out-of-alternation bases that are susceptible to attack by potassium permanganate and thus appear to be significantly unstacked within the left-handed helices. Moreover, the spatial order of successive right- to left-handed DNA transitions within each repeat exhibits a clear polarity toward the distal regulatory region of the rPRL gene. An additional transition involving the long-range unpairing of (A + T)-rich sequences establishes a directional propagation toward the regulatory region. These data demonstrate a complex series of quasi-independent transitions to non-B DNA structures that impinge upon a known regulatory control region. PMID- 8429552 TI - Antitermination of early transcription in phage HK022. Absence of a phage-encoded antitermination factor. AB - In phage lambda and its relatives most early phage genes are located downstream from transcription termination sites, and full gene expression requires suppression of termination (or antitermination). Phage HK022, a lambda relative, also antiterminates early transcription, but, unlike its relatives, does so in the absence of any active phage gene product. We found no functional equivalent of the lambda N antitermination protein in HK022. In addition, nus mutations, which alter host proteins required for lambda antitermination, have no apparent effect on HK022 early gene expression. We have shown that terminators located several thousand base-pairs from the start point of transcription are suppressed, and that in the left operon suppression requires a short, promoter-proximal segment. A 40 bp region within this segment is repeated in the right operon. The chromosomal locations of these repeated segments resemble those of the nut antitermination sites of other lambdoid phages, but the HK022 sites lack the conserved sequence elements of the nut sites. It appears that HK022 antiterminates early transcription in a novel way. PMID- 8429553 TI - Functional sites in the 5' region of human immunodeficiency virus type 1 RNA form defined structural domains. AB - The 5' region of HIV-1 RNA contains functional elements involved in key steps of the retroviral cycle, such as genomic RNA transcription, splicing, translation, dimerization or initiation of reverse transcription. In the present work, we investigated the conformation of the first 500 nucleotides covering the RNA leader and the 5' gag coding sequences of HIV-MAL, using chemical probing. We provide detailed information on almost each nucleotide at one of their Watson Crick positions and on position N-7 of purines. Experiments were conducted on two in vitro transcribed RNA fragments (1 to 707 and 1 to 311). A secondary structure model was derived by combining the experimental data, computer predictions and sequence comparison. Under conditions favoring dimerization (high salt concentration), HIV-1 RNA folds into independent structural domains that can be related to defined functional regions. The first domain corresponds to TAR forming a stable stem-loop. Intrinsic structural features are found to stabilize the TAR hairpin loop. The second domain (nucleotides 56 to 299) contains the PBS sequence, which is located in a stable subdomain constrained by a four stem junction (nucleotides 139 to 218). Although the MAL isolate has an insertion near the PBS, probably resulting from the duplication of a 23-nucleotide sequence, the structural organization of this subdomain is conserved in all other HIV-1 isolates. The third domain (nucleotides 300 to 404) contains the splice donor site, packaging and dimerization elements and the AUG initiation codon of gag. A major result is the structural versatility of this region. Two mutually exclusive structures, both equally in agreement with probing data, could modulate the different functions involving this domain. The reduced accessibility of the gag translational initiation site possibly accounts for the low efficiency of the in vitro translation of the dimer. Finally, the 5' gag coding sequences form a metastable domain. PMID- 8429554 TI - Assembly of the bacteriophage T4 replication machine requires the acidic carboxy terminus of gene 32 protein. AB - The acidic carboxy-terminal 89-amino acid fragment of bacteriophage T4 gene 32 protein was expressed in Escherichia coli to high levels from an inducible plasmid construct. Infection of induced cells by wild-type T4 phage results in impaired phage DNA synthesis. The time at which DNA synthesis begins and the diminution in DNA synthesis rates correlate with the amount of carboxy-terminal peptide that accumulates intracellularly prior to infection. Correspondingly, when induced cells are infected with viable phage containing a small deletion near the carboxy-terminus of 32 protein (delta PR201), the inhibition of phage DNA synthesis was much more severe. The mutant 32 protein competes less well against overproduced wild-type acid peptide than does wild-type 32 protein. The purified acid peptide, when used as the attached ligand for affinity chromatography, binds several T4 proteins from phage-infected cells, including 43 protein (T4 DNA polymerase), Dda protein (a DNA helicase), and UvsX protein (a Rec-like recombination protein). Furthermore, at 50- to 100-fold molar excess of acid peptide over intact 32 protein, phage DNA synthesis was specifically inhibited at the initiation step in an in vitro 5-protein DNA replication experiment. We propose that one or more phage replication proteins are titrated as non-productive protein-protein complexes at a site away from the DNA template. This implies that the carboxy-terminal domain of 32 protein is involved in an obligate step of replication machine assembly when the protein is properly attached to ssDNA in the vicinity of a primer-template junction. The assembly defect we observe is strikingly similar to the repression, or "squelching", of the activity of certain eukaryotic transcriptional activators. PMID- 8429555 TI - Accommodating sequence changes in beta-hairpins in proteins. AB - A systematic study of homologous beta-hairpins in proteins of known structure reveals how insertions and deletions (herein known as indels) in the sequence are accommodated. The study was made for 12 protein families comprising 50 different structures, in which there were 49 independent hairpins. Each hairpin was classified according to its loop length and hydrogen bonding pattern. Most indels were found to occur in the loops and their frequency decreases rapidly with the size of the indel and approximately halves for each extra residue inserted. In very short loops, critical glycines are the primary determinants of loop structure and conversions between the two classic two-residue hairpin loops (with type I' and II' beta-turns) are quite common. Longer insertions are often accommodated by extending the beta-ladder and forming extra hydrogen bonds. There are also several indels that are not accommodated in the loop, but by forming a beta-bulge in one of the strands. This study should provide a useful aid to modelling hairpins in homologous structures. PMID- 8429556 TI - Estimation and use of protein backbone angle probabilities. AB - A procedure is described for estimating the probabilities for the backbone phi psi angles of a protein molecule from the data base of known protein structures. The procedure is basically an adaptation of a published secondary structure prediction scheme, applied to the phi-psi angle bins rather than to the secondary types. The phi-psi angle probabilities estimated this way include all effects of local sequence and are "context sensitive" in that the probabilities for a given residue type depend on its position along the sequence. These probabilities can be used to predict the three-dimensional structure of short polypeptides that are stabilized mainly by local interactions only and to predict the protein folding initiation sites, with moderate to good success rates in each case. They are also potentially useful for efficient sampling in a Monte Carlo scheme of protein tertiary structure prediction methods. PMID- 8429557 TI - Structure of a sarcoplasmic calcium-binding protein from amphioxus refined at 2.4 A resolution. AB - The three-dimensional structure of a sarcoplasmic Ca(2+)-binding protein from the protochordate amphioxus has been determined at 2.4 A resolution using multiple isomorphous-replacement techniques. The refined model includes all 185 residues, three calcium ions, and one water molecule. The final crystallographic R-factor is 0.199. Bond lengths and bond angles in the molecules have root-mean-square deviations from ideal values of 0.015 A and 2.8 degrees, respectively. The overall structure is highly compact and globular with a predominantly hydrophobic core, unlike the extended dumbbell-shaped structures of calmodulin or troponin C. There are four distinct domains with the typical helix-loop-helix Ca(2+)-binding motif (EF hand). The conformation of the pair of EF hands in the N-terminal half of the protein is unusual due to the presence of an aspartate residue in the twelfth position of the first Ca(2+)-binding loop, rather than the usual glutamate. The C-terminal half of the molecule contains one Ca(2+)-binding domain with a novel helix-loop-helix conformation and one Ca(2+)-binding domain that is no longer functional because of amino acid changes. The overall structure is quite similar to a sarcoplasmic Ca(2+)-binding protein from sandworm, although there is only about 12% amino acid sequence identity between them. The similarity of the structures of these two proteins suggests that all sarcoplasmic Ca(2+) binding proteins will have the same general conformation, even though there is very little conservation of primary structure among the proteins from various species. PMID- 8429558 TI - Left-handed polyproline II helices commonly occur in globular proteins. AB - The main-chain conformations of 80 proteins were analysed to identify helical structures that commonly occur but do not fall into the known classes of alpha helix, 3(10)-helix and beta-sheet. The analysis yielded 96 occurrences of four or more sequential residues forming the threefold left-handed poly-L-proline II (PPII) helix. This contradicts the previously held opinion that left-handed helices are rare in globular proteins. The main-chain dihedral angles of these helices form a cluster in phi,psi space that has a maximum at -75 degrees, 145 degrees, corresponding to conformations with the number of residues per turn (n) = -3.0. We show that 51% of PPII-helices lie within the range of n = -3.0(+/- 0.2). It is shown that the PPII segments are distinct from the conformation typical of beta-pleated sheets. Although proline residues commonly occurred in PPII-helices, this side-chain is not obligatory, as 28 of these helices did not contain proline. In addition, we found 120 segments with three C alpha atoms forming a PPII-helix. PPII-helices tend to occur on the surface of the protein and, having few main-chain hydrogen bonds with the rest of the protein, tend to be the more mobile segments of the molecule. The geometry of PPII helix allows the polypeptide chain to progress immediately from this conformation to right handed alpha-helix and 3(10) helix, as well as to beta-sheet or reverse turn. We conclude that PPII-helices should be considered as a regular conformation and should be added to beta-sheets, alpha-helices and 3(10)-helices in databases of protein structures, in secondary structure prediction and in tertiary model building. PMID- 8429559 TI - Domain closure in adenylate kinase. Joints on either side of two helices close like neighboring fingers. AB - In large variants of adenylate kinase the AMP and ATP substrates are buried by a domain rotating by 90 degrees. Here conformational changes responsible for this domain closure are determined by an analysis of the open state of beef heart mitochondrial adenylate kinase and the closed state of Escherichia coli adenylate kinase. Although these two proteins have sequence differences, the principal structural changes responsible for the domain movements are large, and can clearly be distinguished from the effects of evolution. The mobile domain is linked to the rest of the protein by two helices packed together in an antiparallel fashion. During the closure, deformations take place in four localized regions, called joints, near the N and C termini of these helices. Three of these joints have simple motions that can be well approximated by rotations of three torsion angles, but the joint that makes contact with the ligand involves motion throughout an extended loop: i.e. two torsions on either side of a reverse turn change significantly. The main chain atoms of the joints have few packing constraints. The first pair of joints is responsible for approximately 30 degrees of the total rotation and the second pair for the remaining approximately 60 degrees. These movements carries along the regions between the joints, the two helices and the rest of the mobile domain, to a first approximation, as rigid bodies. This jointed domain closure mechanism is contrasted with the shear mechanisms found in other enzymes. PMID- 8429560 TI - Cubic phases of lipid-containing systems. A translational diffusion study by fluorescence recovery after photobleaching. AB - The lateral diffusion coefficient of fluorescent lipid analogues incorporated in four cubic phases of lipid-water systems was determined by the modulated fringe pattern photobleaching technique. In two of the phases, Q230 and Q224, whose structure is bicontinuous, the diffusion is almost as fast as in the fluid lipid bilayers, and is essentially independent of the chemical nature of the probe. In the other two phases, whose structure consists of disjointed hydrocarbon micelles embedded in a water matrix (phase Q223, type I) and of water-containing micelles embedded in a hydrocarbon matrix (phase Q227, type II), the diffusion coefficient is strongly dependent on the chemical structure of the probe and on the topological type (I or II) of the structure. The conclusion is drawn that in the micellar phases the apparent diffusion mirrors the ability of the probe to hop from micelle to micelle. PMID- 8429561 TI - Freeze-fracture electron microscope study of lipid systems. The cubic phase of space group Pm3n. AB - The cubic phase Q223 (space group Pm3n) of lipid-water systems has been studied by freeze-fracture electron microscopy. Four types of fracture planes were identified; all display highly ordered two-dimensional domains, each subdivided into subdomains related to each other by displacements and rotations connected to the symmetry of the space group. The images were filtered using cross-correlation averaging techniques and the filtered images were compared with the corresponding planar sections of the electron density map. The similarity of the two distributions was assessed via a mathematical parameter, the matching index, whose values were determined as a function of the apodization of the electron density map and of the position of the plane in the direction normal to the fracture. By definition, the best coincidence corresponds to an extremum of the matching index. Several conclusions were drawn. (1) The structure of the sample is well preserved in the replicas. (2) The "resolution" of the electron microscope experiments is remarkably close to that of the X-ray scattering study. (3) Of the two types of models that have been envisaged for phase Q223, the micellar is found to be in better agreement with the electron micrographs than the cagelike. (4) The symmetry of the space group is faithfully reflected in the electron microscope images; in particular, the presence of mirror planes rules out the non-centric of the two space groups that are compatible with the crystallographic data. (5) The crystallographic orientations of the fracture planes that are most frequently identified coincide with those of the most intense X-ray reflections; this observation was interpreted as an indication that the fracture propagates differently in the hydrocarbon and in the water volumes. (6) The thinner metal deposits in the replicas were found to coincide with the hydrocarbon regions of the fractures. (7) The fracture seems to be planar across the water matrix and the micelles seem to be removed from the replica and replaced by dips. PMID- 8429562 TI - Cubic phases of lipid-containing systems. Elements of a theory and biological connotations. AB - It has recently been shown that the structure of two of the six cubic phases so far identified in lipid-containing systems is micellar, one (Q223) of type I, the other (Q227) of type II. The micelles of both phases belong to two distinct classes, those of each class being centred at one of the special positions of the space group. From the chemical viewpoint, phase Q227 seems to require a heterogeneous mixture of water-miscible and water-immiscible lipids, whereas phase Q223 has been observed with chemically pure lipids. Also, the area/volume ratio measured at the polar/apolar interface takes the same value in the two types of micelles of phase Q223, different values in those of phase Q227, in keeping with the notion that the area/molecule ratio is closely related to the chemical activity of the lipid components. The topological properties of the micellar phases are profoundly different from those of the bicontinuous phases. The bicontinuous cubic phases (Q230, Q224, Q229) are often presented as paradigms of the infinite periodic minimal surfaces (IPMS). Some authors have generalized that notion and sought in the IPMS a unified theory underlying the entire field of lipid polymorphism. These analogies entertain some confusion between the mathematical concept of surface and the physical notion of interface. A few electron density maps are presented to document the distance that separates the polar/apolar interfaces from the IPMS. The maps also show that some of the geometric singularities (points, lines, surfaces) of the structures coincide with the locus of the CH3 ends of the chains and with the very centre of the water matrix, i.e. with the regions where the short-range disorder is highest. We introduce the expression chaotic zones to designate these regions. In all the lipid phases the chaotic zones are found to occupy special geometric positions, either related to the symmetry elements or to the IPMS. It thus appears that it is energetically more advantageous to adopt an orderly disposal of the short range disorder than to minimize the area of the polar/apolar interfaces. Finally, regarding the possible biological significance of lipid polymorphism, the point is stressed that among the phases that are observed in equilibrium with excess water (these phases are also the most likely candidates for a biological role) those with a cubic symmetry deserve special attention.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429563 TI - Crystallization of pancreatic procolipase and of its complex with pancreatic lipase. AB - The human pancreatic lipase-porcine procolipase complex has been crystallized in space group P3(2)21 (a = b = 80.3 A and c = 251 A) from a solution containing polyethylene glycol, NaCl and beta-octyl glucoside. The crystals diffract to 2.6 A on a synchrotron beam. The complex in the presence of bile salts and phospholipids crystallizes in a tetragonal space group P4(2)2(1)2 (a = b = 133.4 A, c = 92.6 A). Crystals of procolipase alone were obtained under slightly different experimental conditions (space group I432, a = b = c = 164.3 A). PMID- 8429564 TI - Crystallization and preliminary X-ray studies of the basic lectin from winged bean (Psophocarpus tetragonolobus). AB - The basic lectin from winged bean (Psophocarpus tetragonolobus) could be crystallized using polyethyleneglycol (PEG) 4000 (I), PEG 8000 (II) and 2 methylpentane-2,4-diol (MPD) (III) as precipitants. Crystal forms I and II grew in the presence of methyl-alpha-D-galactopyranoside or N-acetylgalactosamine while III grew in the absence of sugar. The three forms have the same space group (P2(1)2(1)2) and similar unit cell dimensions with two dimeric molecules in the asymmetric unit. The unit cell dimensions are a = 156.8 A, b = 89.0 A, c = 73.3 A for I, a = 155.5 A, b = 92.3 A, c = 72.5 A for II and a = 148.3 A, b = 90.7 A, c = 73.8 A for III. The crystals, particularly those grown using PEG 8000, are suitable for high resolution X-ray analysis, which is in progress. PMID- 8429565 TI - Crystallization and preliminary X-ray analysis of the beta-galactosidase from the extreme thermophilic archaebacterium Sulfolobus solfataricus. AB - The beta-galactosidase from the extreme thermophilic archaebacterium Sulfolobus solfataricus has been crystallized from polyethylene glycol 4000 in the presence of sodium acetate and acetate buffer at pH 4.6. The protein crystallizes in P3(1)21 or P3(2)21 (a = 169.4, c = 98.29) and the crystals diffract beyond 2.5 A. The measured crystal density (approximately 1.28 g/cm3) is consistent with the presence of a tetramer (molecular mass 240 kDa) in the asymmetric unit. The specific volume of the crystals is 1.7 A3/Da, indicating a solvent content by volume of only 27%, which is amongst the lowest values observed for protein crystals, and indicates virtual close-packing of the tetramers. PMID- 8429566 TI - Crystallization and preliminary X-ray diffraction studies of beta-cryptogein, a toxic elicitin secreted by the phytopathogenic fungus Phytophthora cryptogea. AB - Crystals of the basic elicitin secreted by Phytophthora cryptogea have been obtained by the hanging-drop method of vapor diffusion from sodium chloride solutions. The crystals belong to the tetragonal space group P4(1)22 (or enantiomorph P4(3)22), with unit cell dimensions a = b = 47 A, c = 137 A and probably contain two molecules per asymmetric unit. The crystals are very stable to X-rays and diffract to 2.2 A resolution on a synchrotron radiation source. PMID- 8429567 TI - Crystallographic characterization of a PHO4-DNA complex. AB - Crystals have been obtained of the DNA-binding domain of the yeast transcription factor PHO4 in complexes with several synthetic fragments of DNA with appropriate cognate sequences. Crystals suitable for X-ray diffraction studies were produced in the case of a complex of the protein with a 17 base-pair fragment of DNA from a solution in polyethylene glycol and calcium chloride. The crystals have the space group of P4(1)2(1)2 or P4(3)2(1)2 with unit cell dimensions a = b = 56.7 A, c = 447.8 A. The diffraction data at 3 A resolution were collected using synchrotron radiation with a Weissenberg camera for macromolecular crystallography. PMID- 8429568 TI - Nucleotide sequence and characterization of Rhizobium meliloti nodulation competitiveness genes nfe. AB - Rhizobium meliloti large plasmid pRmeGR4b carries the nodulation competitiveness locus nfe responsible for the nodulation efficiency and competitive ability of strain GR4 on alfalfa roots. We report here the nucleotide sequence and characterization of a 3345 base-pair DNA section of the nfe region. Sequence analysis revealed four open reading frames (ORFs), two of them with rightward polarity, termed nfe1 and nfe2, are preceded by functional nif consensus sequences and NifA-binding motifs. An additional, NifA-independent, transcriptional start site for nfe1 was also found. Two other ORFs with leftward polarity, designated ORFA and ORFB, were identified upstream from nfe1 and nfe2 but no nif consensus sequences were found. However, expression of ORFA might be indirectly coupled to the NifA-NtrA regulatory network. The gene products of nfe1 and nfe2 were identified using in vitro transcription/translation and bacteriophage T7 RNA polymerase/promoter system, respectively. A high degree of homology between the amino terminal domain of Nfe1 and the nifH gene product was found. In addition, nfe1 shows homology with the upstream non-coding DNA region of the fixABCX operon. Furthermore, the putative ORFB encoded protein contains a helix-turn-helix motif that resembles the DNA-binding consensus sequence proposed for many prokaryotic regulatory proteins. PMID- 8429569 TI - The impact of human immunodeficiency virus on presentation and diagnosis of tuberculosis in a cohort study in Zambia. AB - Two hundred and forty-nine patients with tuberculosis were recruited to a cohort study to investigate the interaction between tuberculosis and HIV in Lusaka, Zambia; findings at presentation are presented here. One hundred and eighty-two (73%; 95% confidence interval 67-79%) of the cases were HIV-1 antibody positive. The diagnosis of tuberculosis was confirmed by microscopy for acid-alcohol fast bacilli, culture of Mycobacterium tuberculosis, or histology in 74% of all cases. HIV negative and positive cases differed in site of disease: among HIV negative patients 72% had pulmonary disease alone, 16% extrapulmonary disease alone and 12% had both, whereas among HIV positive patients 40% had pulmonary disease alone, 34% extrapulmonary disease alone and 26% both (P < 0.001). HIV negative and positive cases were compared with regard to outcome of diagnostic procedures: 55% of HIV negative cases could be diagnosed at enrollment by sputum smear, but only 35% of HIV positive cases (P < 0.01). Among pulmonary cases confirmed by sputum culture, 76% of HIV negative patients had a positive sputum smear, compared with 57% of HIV positive patients (P = 0.09). Pleural and pericardial disease were difficult to confirm, but culture of pleural fluid was positive in 12/46 HIV positive patients, compared with 0/11 HIV negative patients. Lymph node disease was readily confirmed by biopsy. The tuberculin test was positive in only 30/110 (27%) of HIV positive cases, but in 21/38 (55%) of HIV negative cases (P < 0.01). Mycobacterium tuberculosis was cultured in 57% of HIV negative cases and 54% of HIV positive cases; no atypical mycobacteria were isolated. Initial resistance to isoniazid was present in isolates from 5% of cases with a positive culture. PMID- 8429570 TI - Prospects for the use of larvivorous fish for malaria control in Ethiopia: search for indigenous species and evaluation of their feeding capacity for mosquito larvae. AB - Because of problems with drug and insecticide resistance, the National Organization for the Control of Malaria and other Vectorborne Diseases, Ethiopia, has embarked on a programme of research on alternative malaria control methods, including the use of biological control agents, such as larvivorous fish. The objectives of the study were to identify indigenous larvivorous fish species which could be potential candidates for use as biological control agents; to extend knowledge of their distribution in Ethiopia; and to conduct laboratory tests to determine their feeding capacity. An extensive search resulted in the identification of 11 larvivorous fish species indigenous to Ethiopia, including five species previously unrecorded in the country. Seven species were assessed under standard laboratory conditions for their feeding capacity on larvae of Anopheles gambiae s.l. and Culex andersoni. All species tested were efficient larvivores in the laboratory. However, their larvivorous capacity should be tested further in field trials. Based on the findings of this study, two priority areas for the assessment of biological control using larvivorous fish were identified, the port city of Assab, using the local species Aphanius dispar, and the Ogaden, south-eastern Ethiopia, using the local species Oreochromis spilurus spilurus. PMID- 8429571 TI - An experimental study on the effect of praziquantel and oltipraz on some lysosomal enzymes. AB - The in-vivo effect of the schistosomicidal drugs praziquantel and oltipraz on the activities of the liver lysosomal enzymes in Schistosoma mansoni-infected and non infected mice was studied. The effect of S. mansoni infection and the administration of the schistosomicidal drugs on the activities of beta glucuronidase, acid ribonuclease and alpha-naphthyl acetate esterases may be considered as indices for carcinogenicity. Drugs were given orally in subcurative doses, either in a single dose of 400 mg kg-1 for praziquantel or in five daily doses of 50 mg kg-1 oltipraz. The increase in enzymatic activities in infected animals was attributed to deranged metabolic function as a result of liver cell injury. Treatment of uninfected animals with either praziquantel or oltipraz significantly increased the activities of the three lysosomal enzymes. Praziquantel possesses reversible and less toxic effects on the liver than oltipraz. The role of these antischistosomal drugs cannot be ignored as a possible aetiological factor implicated in the process of carcinogenesis associated with schistosomiasis infection through modulation of the operating potential of the enzymes concerned with detoxification, protein and fat metabolism. PMID- 8429572 TI - Frequency and enterotoxigenicity of Campylobacter jejuni and C. coli in domestic animals in Pakistan as compared to Sweden. AB - Domestic and wild animals have been considered to be the most common reservoirs of Campylobacter for human infections. This study has been conducted to establish the frequency and enterotoxigenicity of C. jejuni and C. coli in the stools of domestic animals in a developing versus a developed country. Faecal samples of 300 chickens and laying hens, 102 horses, 180 cows, 92 goats and sheep were collected from Pakistan and Sweden. The hippurate hydrolysis test and CHO-cell test were performed for species differentiation and enterotoxigenicity determination respectively. The pattern and sequence of frequency distribution of Campylobacter in animals in Pakistan and Sweden were similar, the main difference being a significantly higher frequency in Pakistani samples. C. jejuni was the dominant species from both countries and Pakistani strains were more enterotoxigenic than Swedish strains. We conclude that the spread of Campylobacter infection in developing countries originates from different sources in the neighbourhood, for example animals, and unless hygienic conditions improve no change can be expected. PMID- 8429573 TI - Prevalence of leptospiral agglutinins among conservancy workers in Madras City, India. AB - In a study of 584 Corporation conservancy (sanitation) workers who lived mostly in slums, and who worked in four Corporation Circles of Madras City, India, 192 (32.9%) were found to be positive for agglutinins to Leptospira interrogans. Seropositivity prevalence increased with age, but was similar in males and females except in the youngest age group, where males predominated. Prevalence in the four study areas ranged between 17.8 and 40.5% (P < 0.01). Among 152 sera in which one serogroup predominated, Autumnalis was the most commonly recorded (33.6%), followed by Icterohaemorrhagiae (15.1%), Panama (15.1%), Sejroe (14.5%) and others (21.7%). Forty sera reacted to two or more serogroups at the same (highest) titre, most frequently to the first three serogroups above. The titre range was 1:50-1:3200 (geometric mean titre 149). Among a group of 46 male automobile industry workers who lived in middle-class housing, seropositivity prevalence (17.4%) was approximately half that of the sanitation workers (P < 0.05), and the titre range was lower (1:50-1:200, GMT 84). The predominating serogroups were those found in the sanitation workers. Bearing in mind that sanitation workers are the urban group probably at highest risk of leptospiral infection, the prevalence rate (< 33%) found in our study is not considered to be particularly high. PMID- 8429574 TI - Human brucellosis in Dhofar, Sultanate of Oman. AB - Sera were collected, mostly from school children, in six localities of the southern region of the Sultanate of Oman. Macro and micro-agglutination tests were used to indicate positive Brucella serology. Four of the 525 sera tested had titres of at least 1:200, which were considered positive, and two had borderline values. The frequency of serologically positive sera in the six localities ranged between zero and 2%. No relevant difference was observed between titres using antigen of B. abortus or B. melitensis. PMID- 8429575 TI - Resistance of bacteria from human faecal flora to antimicrobial agents. AB - The faecal bacteria from 197 hospital patients, 58 laboratory workers, 66 urban dwellers, and 19 rural dwellers were examined for resistance to five antimicrobial agents (ampicillin, streptomycin, tetracycline, kanamycin, gentamicin). In the majority (68.5%) of faecal samples from people without a recent history of taking antibiotics, 10% or more of the total organisms were resistant to at least one of the antibiotics. This extensive study revealed a high prevalence of resistant bacteria in the gut flora of ambulatory and hospitalized individuals whether or not they were taking antibiotics. PMID- 8429576 TI - Sensitivity of Plasmodium falciparum to pyrimethamine in vivo and to sulphadoxine/pyrimethamine combination in vitro in pregnant women of northern Nigeria. AB - The in-vivo response of Plasmodium falciparum to a single dose of 25 mg pyrimethamine was evaluated in pregnant women in a semi-rural area of Zaria, northern Nigeria. Forty-four women were enrolled in the study and followed up for 14 days. Thirty-one (70%) of the women had cleared their parasitaemia by Day 7 and they remained parasite negative throughout the 14-day period of observation. The MPCT was 2.48 +/- 0.76 days. Thirteen (30%) of the 44 women were parasite positive either on Day 7 (7.16%) or Day 14 (6.14%) and re-treatment with a curative dose of chloroquine (25 mg kg-1 over 3 days) resulted in complete parasite clearance. Of the eight P. falciparum isolates successfully cultured from these women, seven (87.5%) were highly sensitive and one was resistant in vitro to the SDX/PYR combination. PMID- 8429577 TI - Temporomandibular dislocation: a complication of tetanus. AB - A rare complication of tetanus, dislocation of the temporomandibular joint, is presented. The dislocation was extremely difficult to reduce and recurred with repeated spasms. PMID- 8429578 TI - A case of acute aconitine poisoning caused by chuanwu and caowu. AB - A case of aconitine poisoning following the ingestion of 'chuanwu', the main root of Aconitum carmichaeli, and 'caowu', the root of Aconitum kusnezoffii is described. A 35-year-old man became unwell about 90 minutes after ingesting these herbs and his symptoms lasted for about two days. In addition to the typical gastrointestinal (nausea and vomiting) and neurological (generalized weakness, numbness and paraesthesia) features of aconitine poisoning, there was evidence of cardiotoxicity (hypotension and ventricular ectopics). Management of such patients should include measures to enhance elimination and close monitoring for cardiotoxicity. Legislation controlling the use of such herbs should be considered. PMID- 8429579 TI - Adult respiratory distress syndrome (ARDS) complicating typhoid fever. PMID- 8429580 TI - Effect of iodophor vs iodine tincture skin preparation on blood culture contamination rate. AB - OBJECTIVE: To determine if iodine tincture is a more effective skin antiseptic for blood culture collection than povidone-iodine, an iodophor. DESIGN: Pairwise comparisons across phases. In phases 1 and 3, blood culture skin preparation was performed with the iodophor; in phases 2 and 4, skin preparation was performed with iodine tincture. SETTING: Emergency department of a large urban teaching hospital. PATIENTS: All adult patients seen in the emergency department who had blood cultures collected because a systemic bacterial infection was suspected. MAIN OUTCOME MEASURE: The blood culture contamination rate for the iodophor and iodine tincture skin preparations. RESULTS: A total of 8467 blood cultures were collected during the study, and 421 (4.97%) were classified as contaminated. The contamination rate for the blood cultures collected using the iodophor was 6.25%, and the contamination rate for the cultures using iodine tincture was 3.74%; this difference is statistically significant (P < .00001). CONCLUSIONS: The effectiveness of the skin antiseptic may be an important factor in determining contamination rate in blood culturing. If these results are confirmed by others, then institutions that have a high blood culture contamination rate when using an iodophor for skin preparation should consider changing to iodine tincture. PMID- 8429581 TI - The accuracy of predictions of violence to others. AB - OBJECTIVE: To assess the accuracy of clinicians in predicting violence in mental patients. Specifically, to determine if clinicians can predict violence when variation in rates of violence attributable to age, race, and sex is controlled. DESIGN: Two samples of psychiatric patients, matched on age, race, sex, and admission status, were followed up in the community during a 6-month period. One group included individuals assessed by psychiatric emergency department clinicians as likely to be violent to another person during the follow-up period; the other was a comparison group. Patients provided self-reports of violent incidents, and a "collateral," ie, an individual with detailed knowledge of the patient's life, provided this same information. Official records were also reviewed. SETTING: Patients were recruited in the emergency department of a metropolitan psychiatric hospital. Patients and collaterals were interviewed in their homes or in public places in the community. PATIENTS OR OTHER PARTICIPANTS: A consecutive sample of individuals coming into a psychiatric emergency department during daylight and evening shifts was obtained. A total of 2452 patients were approached for consent and 1948 consented. A final sample of 357 patients whom clinicians assessed as likely to be violent and their matched comparison patients were included. MAIN OUTCOME MEASURES: Patients', collaterals', and official records' reports of incidents in which the patient laid hands on another person or threatened someone with a weapon. RESULTS: Violence during the follow-up period was reported in approximately 45% of the cases: 36% in the comparison group and 53% in the cases predicted to be violent. Overall clinical accuracy was significantly better than chance, but predictions of female patients' violence were not better than chance. CONCLUSIONS: The level of patient violence reported using self-reports and collateral reports was higher than has been obtained using other methods. Clinical judgment adds to predictive accuracy, but overall accuracy was modest and particularly low for female patients. PMID- 8429582 TI - The era of the patient. Using the experience of illness in shaping the missions of health care. AB - The emergence of an era that focuses on the experiences of individuals with illness to provide an alternative voice in health care is explored. Antecedent events that caused the eclipse of the patient and challenged the authenticity of personal experience in establishing medical facts--the introduction of the concept of diseases and the technologic revolution in medicine--are examined. Recent events that returned the patient's views to the center of medical attention--the medical ethics and outcomes movements--are analyzed and connected. Recommendations are presented to make the experience of individuals with illness significant features of health care practice, education, research, and policy, and to reverse the view of patients and subjects as individuals benefited by medicine, but unable to help it. PMID- 8429583 TI - NIH Consensus conference. Gallstones and laparoscopic cholecystectomy. PMID- 8429584 TI - Mechanical ventilation for the elderly patient in intensive care. Incremental changes and benefits. AB - OBJECTIVE: To evaluate the cost-effectiveness of prolonged mechanical ventilation in patients 80 years of age and older in the intensive care unit (ICU). DESIGN: A retrospective review of consecutive ICU patients requiring 3 or more days of mechanical ventilation. Cost-effectiveness analysis was performed by assessing incremental hospital charges from hospital billing records; charges were then related to years of life saved. A telephone survey was used to follow up hospital survivors for a minimum of 4 years after discharge. SETTING: A 20-bed medical surgical ICU in a 420-bed, tertiary-care community teaching hospital. PATIENTS: The study included all patients aged 80 years or older taken from a comprehensive database of all patients admitted to the ICU requiring mechanical ventilation from April 1, 1985, through October 31, 1987 (n = 512). Of 59 potential candidates, 45 were found to have complete billing records and were the subject of further analysis. RESULTS: Of the 45 patients in the group under analysis, 10 survived to leave the hospital. Of these, two were alive and one could not be located at the time of follow-up. The charge per year of life saved is estimated to between $51,854 and $75,090 in 1985-1987 dollars. Of 22 patients whose age in years plus duration of mechanical ventilation in days totaled 100 or greater, only two survived hospitalization and neither was alive at follow-up. The cost per year of life saved in this subset of patients was $1181,308 in 1985-1987 dollars. One of these patients was discharged to a nursing home and died there 4.5 years later, after multiple hospital readmissions. The other patient died at home 2 months after hospital discharge. CONCLUSION: Based on hospital charges and life expectancy, the cost-effectiveness of prolonged mechanical ventilation in ICU patients age 80 years and over was poor in our population when the combination of age and duration of mechanical ventilation exceeded 100. Further studies using this type of analysis may prove valuable in both clinical and administrative decision-making processes. PMID- 8429585 TI - Perceptions and misperceptions of race and drug use. PMID- 8429586 TI - Is baldness bad for the heart? PMID- 8429588 TI - A piece of my mind. The messengers. PMID- 8429587 TI - The double helix--Watson & Crick's 'freak find' of how like begets like. PMID- 8429589 TI - Women physicians target barriers. PMID- 8429590 TI - After two decades, Penn State researchers may be near perfecting replacement hearts and lungs. PMID- 8429591 TI - From the Food and Drug Administration. PMID- 8429592 TI - From the Centers for Disease Control and Prevention. Sexual risk behaviors of STD clinic patients before and after Earvin 'Magic' Johnson's HIV-infection announcement--Maryland, 1991-1992. PMID- 8429593 TI - From the Centers for Disease Control and Prevention. Probable transmission of multidrug-resistant tuberculosis in correctional facility--California. PMID- 8429594 TI - From the Centers for Disease Control and Prevention. Ceftriaxone-associated biliary complications of treatment of suspected disseminated Lyme disease--New Jersey, 1990-1992. PMID- 8429595 TI - Donation of unused surgical supplies: help or hindrance. PMID- 8429596 TI - Donation of unused surgical supplies: help or hindrance. PMID- 8429597 TI - Donation of unused surgical supplies: help or hindrance. PMID- 8429598 TI - Subselecting patients at risk for pulmonary embolism. PMID- 8429599 TI - Mammography after augmentation mammoplasty. PMID- 8429600 TI - Truth telling: a cultural or individual choice? PMID- 8429601 TI - Truth telling: a cultural or individual choice? PMID- 8429602 TI - Laparoscopic adrenalectomy for primary aldosteronism. PMID- 8429603 TI - The postmenopausal estrogen/breast cancer controversy. PMID- 8429604 TI - The postmenopausal estrogen/breast cancer controversy. PMID- 8429605 TI - Probing the meaning of racial/ethnic group comparisons in crack cocaine smoking. AB - OBJECTIVE: To probe the meaning of reported racial and ethnic group differences in the prevalence of crack cocaine smoking and to estimate the degree to which crack cocaine smoking is associated with personal factors specific to race/ethnicity. DESIGN: Through reanalysis of data from the 1988 National Household Survey of Drug Abuse (NHSDA), we compared racial/ethnic group differences in crack cocaine smoking. To hold constant social and environmental risk factors that might potentially confound racial comparisons, we used an epidemiologic strategy that involves poststratification of respondents into neighborhood risk sets. A conditional logistic regression model was used to estimate the relative odds of crack use by race/ethnicity. PATIENTS OR OTHER PARTICIPANTS: The 1988 NHSDA interviewed 8814 individuals residing within households in the United States. Subjects were selected using a multistage area probability sampling of all residents aged 12 years and older. RESULTS: Once respondents were grouped into neighborhood clusters, the relative odds (RO) of crack use did not differ significantly for African Americans (RO, 0.85; 95% confidence interval [Cl], 0.37 to 1.93) or for Hispanic Americans (RO, 0.88; 95% Cl, 0.47 to 1.67) compared with white Americans. CONCLUSION: Findings of race associated differences are often presented as if a person's race has intrinsic explanatory power. This analysis provides evidence that, given similar social and environmental conditions, crack use does not strongly depend on race-specific (eg, biologic) personal factors. Although the study finding does not refute the previous analysis, it provides evidence that prevalence estimates unadjusted for social environmental risk factors may lead to misunderstanding about the role of race or ethnicity in the epidemiology of crack use. Future research should seek to identify which characteristics of the neighborhood social environment are important and potentially modifiable determinants of drug use. PMID- 8429606 TI - A case-control study of baldness in relation to myocardial infarction in men. AB - OBJECTIVE: To examine the relationship between male pattern baldness and the risk of myocardial infarction in men under the age of 55 years. DESIGN AND PARTICIPANTS: A hospital-based, case-control study was conducted in eastern Massachusetts and Rhode Island. Cases were men admitted to a hospital for a first nonfatal myocardial infarction (n = 665); controls were men admitted to the same hospitals with noncardiac diagnoses (n = 772). Extent of baldness was assessed using the 12-point modified Hamilton Baldness Scale; other information was obtained by personal interview. Among the controls, the prevalence of any baldness was 34%, while the prevalence of baldness involving the vertex scalp was 23%. RESULTS: After allowing for age, the relative risk estimate for frontal baldness compared with no hair loss was 0.9 (95% confidence interval, 0.6 to 1.3), for baldness involving the vertex scalp it was 1.4 (95% confidence interval, 1.2 to 1.9). Risk of myocardial infarction increased as the degree of vertex baldness increased (P < .01); for severe vertex baldness the relative risk was 3.4 (95% confidence interval, 1.7 to 7.0). The relationship between vertex baldness and myocardial infarction was consistent within strata defined by age and other risk factors for coronary artery disease. CONCLUSION: These data support the hypothesis that male pattern baldness involving the vertex scalp is associated with coronary artery disease in men under the age of 55 years. PMID- 8429607 TI - Membership in ANA: a component of professionalism. PMID- 8429608 TI - Nursing: profession or occupation? PMID- 8429609 TI - All ethics problems are not created equal. PMID- 8429610 TI - Vision for the future. PMID- 8429611 TI - [1993: European year of the aged and of intergenerational solidarity]. PMID- 8429613 TI - [Geriatric care in Hungary]. PMID- 8429612 TI - [Geriatric care in Switzerland]. PMID- 8429614 TI - [Voices from the geriatric nursing congress. Interview by Peter F. Meurer]. PMID- 8429615 TI - [A device aiding patients to sit up. A critical view]. PMID- 8429616 TI - [Attempted suicide--suicide--prevention. A nursing viewpoint]. PMID- 8429617 TI - [Publishing house and editorial staff of "Krankenpflege"]. PMID- 8429618 TI - [Legal accident insurance protection in the care of the disabled]. PMID- 8429619 TI - Estimation of perimortal percent carboxy-heme in nonstandard postmortem specimens using analysis of carbon monoxide by GC/MS and iron by flame atomic absorption spectrophotometry. AB - In decomposed, formalin-fixed, embalmed, exhumed, and some fire-dried cases in which normal blood is unavailable, the usual methods for determination of carboxyhemoglobin saturation frequently fail. To address these specimens, a method utilizing both gas chromatography/mass spectrometric (GC/MS) determination of carbon monoxide (CO) and flame atomic absorption spectrophotometry (FAAS) determination of iron (Fe), in the same specimen, was developed. The method is reported here, along with its application to seven pertinent forsensic death investigations. The CO analytical methodology involves acid liberation of the gas from the specimen aliquot in a headspace vial. After heating and equilibrating, a sample of the headspace vapor is injected into the GC/MS system with a gastight syringe. Quantitation is achieved by standard addition comparison utilizing the ideal gas law equation. Iron is quantified by FAAS analysis of the same aliquot used for the CO determination, following nitric acid digestion. The concentration is determined by comparison to a standard curve. A formula for determining the minimum percent carboxy-heme saturation was derived by using the ratio of the amount of CO to the amount of Fe in the aliquot analyzed. Tissue types analyzed include spleen, liver, muscle, dried blood, and unspecified decomposed tissue. PMID- 8429620 TI - The effects of adulterating agents on FPIA analysis of urine for drugs of abuse. AB - A variety of chemical agents were evaluated to determine their effects on fluorescence polarization immunoassays for drugs of abuse. Sixteen different agents, at concentrations up to 10%, were tested against urine assays for cannabinoids, cocaine (metabolite), amphetamines, opiates, phencyclidine (PCP), and barbiturates. The potential to cause both false positive and false negative results was evaluated, and assays were performed one and seven days after sample adulteration to simulate different collection/testing formats. All six drug assays were susceptible to one or more adulterating agents, but the degree varied considerably between assays. The cannabinoid assay was most susceptible to adulterant-induced false negative results, and the barbiturate assay was most susceptible to false positive results. The remaining assays demonstrated relatively few, but characteristic effects, some of which were attributable to drug degradation and others to assay interference. Although the results of pH measurement on adulterated samples verified its utility in identifying some samples adulterated with interfering agents, other adulterants that cause substantial effects would not be identified by pH measurements alone. PMID- 8429621 TI - Preconcentration of heavy metals in urine and quantification by inductively coupled plasma atomic emission spectrometry. AB - This paper describes a method for the determination of heavy metals (Co, Ni, Cu, Cd, Pb) in urine by inductively coupled plasma atomic emission spectrometry (ICP AES). The method proposed requires purification of the samples with activated charcoal under acidic conditions before preconcentration by complexation with ammonium pyrrolidinedithiocarbamate (APDC). The formed complexes are extracted with methyl isobutyl ketone (MIBK) and the resulting residue is finally digested under acid oxidant conditions. Because of its low detection limit (below 10 micrograms/L), this procedure can be applied conveniently for toxicological diagnostic purposes. PMID- 8429622 TI - Fluorescence polarization immunoassay detection of amphetamine, methamphetamine, and illicit amphetamine analogues. AB - The Abbott Diagnostics Amphetamine/Methamphetamine II and Amphetamine Class reagents were evaluated on the Abbott TDx for cross-reactivity to amphetamine and methamphetamine stereoisomers, several of their metabolites, and various illicit analogues, including 2-methoxyamphetamine, 4-hydroxymethamphetamine, 2,5 dimethoxy-amphetamine (DMA), 4-bromo-2,5-dimethoxyamphetamine (DOB), 4-bromo-2,5 dimethoxy-beta-phenethylamine (BDMPEA), 3,4,5-trimethoxyamphetamine (TMA), 3,4 methylenedioxy-amphetamine (MDA) N,N-dimethyl-3,4-methylenedioxy-amphetamine, N hydroxy-3,4-methylenedioxyamphetamine (N-OH MDA), 3,4 methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), 2,5-dimethoxy-4-ethylamphetamine (DOE), 2,5-dimethoxy-4-methylamphetamine (DOM), and mescaline in concentrations ranging from 100 to 100,000 ng/mL. Results demonstrate the utility of this assay for detection of several of the above compounds; unfortunately many are still not detectable. Significant differences were observed between the Amphetamine/Methamphetamine II and Amphetamine Class reagents, particularly regarding their cross-reactivity to over-the-counter medications. Detection of the drugs amphetamine, methamphetamine, and the illicit analogues is not enhanced with the Amphetamine Class reagents, and unless detection of the over-the-counter compounds is of interest, these reagents are a poor choice compared to the Amphetamine/Methamphetamine II reagents. Cross reactivity of some of the illicit analogues is such that the assay can reliably be used for the routine screening of these compounds. PMID- 8429623 TI - Retrospective analysis of some L-methamphetamine/L-amphetamine urine data. AB - A limited analysis is presented of data accumulated over about three years for specimens containing either nonracemic L-methamphetamine/L-amphetamine or racemic mixtures of the D and L stereoisomers. 55 data points for both nonracemic L methamphetamine and racemic mixtures show that it is possible to report a specimen containing L-methamphetamine positive for illegal D-methamphetamine based on current guidelines unless chiral assays are performed on selected methamphetamine positives. Generation of only 45 racemic specimens from chiral analysis of about 5,000 urine methamphetamine positives demonstrates the overwhelming prevalence of nonracemic illegal D-methamphetamine in Southern California. PMID- 8429624 TI - Evaluation of the Syva ETS Plus urine drug and serum ethanol analyzer. AB - The Syva ETS is an automated system designed for analysis of drugs of abuse in urine. Recently, the Syva Company has upgraded this system to the ETS Plus, which is capable of analyzing ethanol in urine, serum, or plasma. We evaluated the new ETS Plus for urine screening of barbiturates, benzodiazepines, cocaine metabolite, opiates, and phencyclidine. Results of 505 patient sample assays obtained by ETS Plus were compared with those of ETS. There were only four discrepant results which had absorbance rates close to the low calibrator cutoff values. The within-run precision of the ETS Plus Ethyl Alcohol Assay yielded a CV of 2.6% at a target value of 1.00 g/L (1.06 +/- 0.03 g/L, n = 32) and a CV of 3.2% at a target value of 0.40 g/L (0.42 +/- 0.01 g/L, n = 30). The linear regression analysis of 30 patient serum ethanol results by the ETS Plus and by gas chromatography yielded y = 0.939 x + 0.03 g/L. The software modifications in the new ETS Plus allow the accurate quantitation of ethanol in serum and reliable detection of drugs of abuse within the same batch. PMID- 8429625 TI - HPLC determination of rubber septum contaminants in the iodinated intravenous contrast agent (sodium iothalamate). AB - A simple and rapid method is described for the determination of 2 mercaptobenzothiazole and mercaptobenzothiazole disulfide leached from rubber closures into the iodinated contrast medium (sodium iothalamate). The contaminants were extracted with methylene chloride and analyzed by high performance liquid chromatography using ultraviolet detection at wavelengths of 272 and 328 nm. The structures of the impurities were confirmed unequivocally by mass spectrometry. The minimum detectable level was 1 microgram/100 mL (0.01 ppm). The quantitation of contaminants leached from rubber stoppers provides a means for evaluating possible health risks of the extractables. PMID- 8429626 TI - Determination of 1-hydroxypyrene in human urine by high-performance liquid chromatography. AB - A high-performance liquid chromatography (HPLC)/fluorescence method for quantitative analysis of 1-hydroxypyrene in urine was developed. The method validation analysis showed the method to be in analytical control. No significant systematical errors could be demonstrated. The entire run time of chromatography was 10 min using isocratic elution (acetonitrile-water, 70:30), and the retention time for 1-hydroxypyrene was 3.5 min. The short run time in combination with the low limit detection (1.37 nmol/L) makes the method potentially applicable for surveillance of pyrene exposure in work environments. The developed method is presently used for measurement of 1-hydroxypyrene in urine samples from workers exposed to a low airborne level of polycyclic aromatic hydrocarbons, generally less than 25 micrograms/m3. The urine samples of exposed workers (n = 122) showed a range of 1-hydroxypyrene from the limit of detection (LD) up to 39 nmol/L, whereas unexposed control individuals (n = 108) showed a range from LD to 3.3 nmol/L. PMID- 8429627 TI - Radioimmunoassay screening of dried blood spot materials for benzoylecgonine. AB - Residual samples of blood spots, which are routinely collected on almost all newborns in the United States, can be used to determine seroprevalence information on newborns and maternal exposures to various substances, including drugs of abuse. By modifying a commercial radioimmunoassay (RIA) kit for urinary samples, one can use blood spotted on filter paper as a matrix to quantitate the cocaine metabolite benzoylecgonine (BE). BE is stable for long periods of time in blood spots and we were able to quantitatively extract it with aqueous buffer. There were no matrix effects of the blood spot eluate on the RIA, and excess lipid in the blood did not alter measurement of BE. By using standards made up of BE in negative blood spot eluate and calibrators of blood that were spiked with BE and then spotted on filter paper to determine extraction efficiency, low levels of BE in blood could be measured. The limit of detection was 5 ng/mL, and the limit of quantitation was 10 ng/mL. Levels of BE in blood collected at autopsy in eluates of blood spots were measured, and they established excellent correlation (r2 = 0.93) with gas chromatography/mass spectrometry measurements. To test this technology, residual blood spots on 545 infants from three states were analyzed for BE. PMID- 8429628 TI - Determination of the heroin metabolite 6-monoacetyl-morphine in urine by high performance liquid chromatography with electrochemical detection. AB - A procedure has been developed for detection and quantitation of 6 monoacetylmorphine (6-MAM), the specific metabolite of heroin in human urine. 6 MAM was isolated by solid-phase extraction, and the extracts were analyzed by high-performance liquid chromatography with electrochemical detection in the oxidizing mode. Good linearity was obtained in the concentration range 10-250 ng/mL. Concentrations of 2 ng 6-MAM/mL can be detected using a 2-mL sample of urine. The method has been verified in urine samples of heroin abusers. PMID- 8429629 TI - Determination of cocaine and benzoylecgonine by derivatization with iodomethane D3 or PFPA/HFIP in human blood and urine using GC/MS (EI or PCI mode). AB - A method for the simultaneous detection and quantification of cocaine and benzoylecgonine from human blood and urine has been developed. Cocaine and benzoylecgonine were extracted from blood and urine with solid-phase columns containing C18 packing material. Clean, emulsion-free extracts were obtained. The analytes were derivatized with PFPA/HFIP or iodomethane-D3. The underivatized cocaine and derivatized benzoylecgonine were determined by GC/MS with positive chemical ionization (PCI) and the selected ion monitoring mode. Benzoylecgonine was derivatized with iodomethane-D3 to cocaine-D3. Both derivatives were chromatographed on a CP-SIL 5 capillary column. The recovery of extraction was greater than 70%. The detection limit for the PFPA/HFIP derivatives (20 ng/mL) was better than that for iodomethane-D3 derivatives (40 ng/mL). The procedure is simple, rapid, and reproducible. Standard curves were linear and the procedure was suitable for routine analysis. Using fluorescence polarization immunoassay (TDx, Abbott) (cutoff 100 ng/mL), along with one of the GC/MS methods described, we found no false positive or false negative results. PMID- 8429630 TI - Simultaneous determination of eight anticoagulant rodenticides in blood serum and liver. AB - A liquid chromatographic method was developed for the analysis of indandione and 4-hydroxycoumarin anticoagulant rodenticides in blood serum and liver. The method enabled the measurement of serum and liver concentrations of eight anticoagulant rodenticides: brodifacoum, bromadiolone, chlorophacinone, coumafuryl, coumatetralyl, diphacinone, difenacoum, and warfarin. Anticoagulants were extracted from serum and liver with acetonitrile. Extracts were applied to solid phase extraction columns, which contained mixed packings. Column eluates were evaporated to dryness, reconstituted, and subjected to reversed-phase liquid chromatography. Hydroxycoumarins were detected by fluorescence at an excitation wavelength of 318 nm and an emission wavelength of 390 nm. Indandiones were detected by UV absorption at 285 nm. Extraction efficiencies of greater than 75% for serum and greater than 69% for liver were obtained. The within-run precision (CV) ranged from 2.4 to 8.6% for serum and 2.6 to 8.7% for liver. The between-run precision (CV) ranged from 1.5 to 12.2% for serum and from 2.1 to 11.8% for liver. Hydroxycoumarin rodenticides were detected at 1 ng/mL of serum and 1 ng/g of liver. Indandiones were detected at 10 ng/mL of serum and 10 ng/g of liver. PMID- 8429631 TI - Determination of cocaine and its major metabolite, benzoylecgonine, in amniotic fluid, umbilical cord blood, umbilical cord tissue, and neonatal urine: a case study. PMID- 8429632 TI - Distinguishing between ethanol and acetonitrile using gas chromatography and modified Widmark methods. PMID- 8429633 TI - Cellular responses to silicone and polyurethane prosthetic surfaces. AB - Prosthetic devices composed of silicone or polyurethane are commonly used in surgery. These devices elicit a soft tissue reaction which may frequently be complicated by capsule formation. Histologically the capsule comprises both cellular (fibroblasts and endothelial cells (EC)) and matrix components (predominantly collagen type I). We hypothesized that the function of the cellular elements is altered by exposure to prosthetic materials and that this alteration contributes to capsule formation. To test this hypothesis, we utilized specific in vitro assays of cell function (attachment, proliferation, matrix gel contraction), which closely mimic in vivo cellular events, in order to define the responses of EC and fibroblasts to prosthetic surfaces (foam polyurethane, flat silicone, and textured silicone). Morphologic changes were evaluated by scanning electron microscopy (SEM). Attachment of both cell types to all prosthetic surfaces was significantly decreased compared to control (HUVEC: control, 55 +/- 1; foam polyurethane, 19 +/- 4*; flat silicone, 25 +/- 3*; textured silicone, 36 +/- 1*; fibroblast: control, 93 +/- 6; foam polyurethane, 21 +/- 4*; flat silicone, 57 +/- 5*; textured silicone, 44 +/- 5* (*P < 0.05 = significant; units, percentage spread)). Fibroblast proliferation was significantly decreased on foam polyurethane (0.1 +/- 0.03*) and textured silicone (0.18 +/- 0.05*), but not on flat silicone (0.79 +/- 0.2; control = 0.96 +/- .2). In contrast, HUVEC proliferation was significantly decreased on both silicone surfaces but not on polyurethane (units, cpm/cell; control, 0.26 +/- 0.05; foam polyurethane, 0.15 +/ 0.05; flat silicone, 0.08 +/- 0.03*; textured silicone, 0.02 +/- 0.01*).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429634 TI - Correlation of high-speed tensile strength with collagen content in control and lathyritic rat skin. AB - Severity of lacerative skin injury depends on the applied load and the resistance of the tissue. At low (static) rates of loading there is a high degree of correlation between skin tensile strength and the degree of collagen crosslinking, with little added strength due to collagen interactions with the glycosaminoglycan matrix. We examined the effects of high (ballistic) rates of loading in order to determine the contributions to strength made by both the degree of collagen crosslinking and the collagen-matrix interaction. Tensile failure experiments were conducted using the dorsal skin of rats 1.5-6 months of age. Test specimen orientations were cut parallel and transverse to the body axis at cephalad and caudad locations on the dorsum. Tensile strength was measured at nominal strain rates of 30%/sec (low speed) and 6000%/sec (high speed) using both control and lathyrogen fed rats. Biochemical analyses were conducted to determine the amount of total and crosslinked (insoluble) collagen. In low-speed tests, there was a significant correlation (r > or = 0.900) between collagen content and skin tensile strength measured both transverse and parallel to the spine. The degree of correlation was higher with insoluble (r = 0.973) collagen content than with total (r = 0.901) collagen. The effect of a lathyrogen diet produced a significant (P < 0.001) reduction (two- to threefold) in tensile strength compared to control. In both high- and low-speed groups, tensile strength was greatest in the transverse samples with a significant correlation to collagen content (r > or = 0.858).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429635 TI - Endovascular canine anastomotic stenting. AB - The healing characteristics and morphological features of a trans-anastomotic endovascular multifilament flexible metallic stent (Wallstent, Schneider) were investigated in a canine end-to-end arterial model. Both femoral arteries of 12 conditioned mongrel dogs were exposed, then transversely divided and reanastomosed. One anastomosis was randomly chosen for stent placement. Stents were chosen to match the diameter of the femoral arteries and were 25 mm in unconstrained length. Animals were sacrificed at intervals of 3, 6, 9, and 12 weeks. The anastomotic segments were pressure perfusion fixed and sectioned. The cross-sectional luminal, intimal, and medial areas were calculated from computer digitized images of the individual sections. Data were analyzed by univariate repeated measures analysis of variance. Stented anastomoses had an increased luminal area as compared with the anastomosis of non stented sections (P < 0.002). However, stended segments had a significantly increased intimal area when compared to control segments in the same animal (P < 0.001). Stented anastomoses had a significantly increased lumen area over controls despite a marked intimal response to the stent. PMID- 8429636 TI - Prevention of postischemic injury in immature intestine by deferoxamine. AB - Free radical-mediated reperfusion injury has been demonstrated in ischemic neonatal bowel necrosis, but the mechanism of injury remains elusive. To determine whether such an injury can be prevented, 76 weaning rats were studied to test the effects of deferoxamine, an iron chelator, in postischemic injury. Group I (N = 20) had a sham laparotomy without vascular occlusion. Group II (N = 21) was subjected to 90 min of superior mesenteric artery occlusion prior to reperfusion. Group III (N = 35) received deferoxamine 15 mg/kg intravenous prereperfusion, in addition to ischemia and reperfusion as in group II. Survival profiles for each group were determined and a scale of pathologic severity was applied and compared. Group I had 100% long-term survival and group II, 14%. Group III had an overall survival of 28% and demonstrated a prolonged postreperfusion survival profile (P < 0.002) compared to group II. Histology was nearly identical to human necrotizing enterocolitis in degrees of bowel wall destruction and relative paucity of neutrophils. Group III showed a significant reduction in severity of injury compared to group II (P < 0.003). We conclude that neonatal bowel ischemia conditions such as necrotizing enterocolitis may be reperfusion injuries wherein free iron plays an important role in tissue injury. Administration of an iron chelating agent under such conditions has a beneficial effect on survival and histology. PMID- 8429637 TI - The effects of tumor necrosis factor after removal of the gut. AB - Tumor necrosis factor (TNF) is thought to be an important mediator of the septic response; intravenous administration of this cytokine provokes a wide range of metabolic and hemodynamic effects which closely resemble those known to follow endotoxin administration. These effects include damage to gut mucosa. Since the gut constitutes an internal reservoir of endotoxin and bacteria it is possible that some of TNF's effects might be secondary to an increased absorption of these agents. To test this hypothesis TNF was given both to intact dogs and also to animals in which the gut had been removed, from duodenum to anus ("enterectomy"). The data indicates that the effects of TNF upon systemic pressure, white count, and cortisol and glucagon levels were not affected by prior removal of the gut. For most other parameters the findings were less clear but in general the changes in enterectomized dogs were qualitatively similar to those described in the intact animal. We conclude that the effects observed following TNF infusion in the dog are not dependent on the intestinal tract as a secondary source of bacteria or mediators. PMID- 8429638 TI - The effect of high energy shock waves focused on cortical bone: an in vitro study. AB - Extracorporeal shock wave lithotripsy has become an accepted alternative for the management of nephrolithiasis and cholelithiasis. Direct impact of shock waves cause tear and shear forces at transition sites between tissues with divergent acoustic impedances leading to stone fragmentation. The aim of this study was to determine whether shock waves can cause cortical bone damage at all and, if so, what the relationship is, if any, between the energy density of the shock waves, the number of shock waves applied, and the resulting cortical bone damage. With the Siemens Lithostar Plus with overhead module, electromagnetic shock waves, generated under water with energy densities of 0.23, 0.33, 0.42, or 0.54 mJ/mm2, corresponding with power settings 2, 4, 6, and 8, were applied to bone specimens, i.e., of rabbit femurs and tibiae. Prior to exposure to the shock waves, the bones were mounted on a specially constructed perspex holder which could be placed in a water-filled test basin with an elastic membrane in the front through which the shock waves propagate without loss of energy. This setup made it possible not only to induce complete fractures, but also to detect the existence of a linear relationship with a Spearman rank correlation coefficient of -0.72 (P < or = 0.01) between the energy level of the shock waves and the severity of the cortical bone defects. The latter findings are especially of great importance because this means that the process can be controlled and that the cortical effects will be predictable and reproducible. This study should be considered a preliminary test concerning the effects of high energy shock wave on bone.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429639 TI - Bioimpedance hemodynamics compared to pulmonary artery catheter monitoring during orthotopic liver transplantation. AB - Simultaneous cardiac output measurements were obtained using bioimpedance (BI) and thermodilution (TD) methods in 10 patients undergoing orthotopic liver transplantation (OLT). BI and TD data were collected during five intraoperative phases of OLT. Linear regression revealed significant correlation between BI and TD (r = 0.8, P = 0.001) with a slope of regression of 0.64. Bias analysis demonstrated a negative of BI (-0.26) when compared with TD.Sv-O2 correlated poorly with cardiac index determined by TD (r = 0.41) or BI (r = .47). BI is an accurate reproducible method for measuring cardiac output and may be a useful adjunct to the pulmonary artery catheter for hemodynamic monitoring during OLT. PMID- 8429640 TI - Effects of glutaraldehyde on experimental arterial iso- and allografts in rats. AB - The effects of glutaraldehyde pretreatment and allograft rejection in arterial grafts were assessed, using iso- and allografts in rats. An in situ glutaraldehyde fixation procedure was used to obtain homogeneous cross-linked vascular biografts. Ten Lewis rats were isografted, ten were isografted with a glutaraldehyde-treated aortic segment, ten were allografted with aortic segments from brown Norway (BN) inbred rats, and ten were allografted with glutaraldehyde treated BN aortas. The macroscopic and microscopic appearances of the grafts were analyzed 3 weeks after the initial surgery. Immunological injury to the media and the intimal response were quantified morphometrically after monochromatic staining of cell nuclei (hematoxylin after periodic acid), elastin (orcein), and calcification (Von Kossa). Untreated isografts were normal. Untreated allografts showed the classical signs of arterial wall rejection: adventitial inflammatory granuloma, reduced medial thickness and smooth muscle cell density, and greatly increased intimal thickness (P < 0.005). Glutaraldehyde treatment significantly decreased the medial thickness in both iso- and allografts (P < 0.001) and prevented the intimal proliferative response (P < 0.005), but did not change adventitial inflammation. It also induced massive calcification mainly in isografts (P < 0.001). Histomorphological modifications of glutaraldehyde-treated grafts are consistent with a partial protective effect of glutaraldehyde against the rejection process, but also with an induction of a nonspecific inflammatory reaction. Glutaraldehyde-induced cross-linking of the extracellular matrix was responsible for ectopic calcification of the arterial grafts which was independent of the rejection process. PMID- 8429641 TI - Reduction of atherosclerosis with nonsteroidal anti-inflammatory drugs. AB - The cellular mechanisms involved in the pathogenesis of atherosclerosis may be similar to the events in the inflammatory response. Many of these processes are activated by the arachidonic acid cascade. The nonsteroidal anti-inflammatory drugs inhibit the cycloxegenase and possibly the lipoxegenase pathways, decreasing the production of prostaglandins, thromboxanes, and leukotrienes, all components of the inflammatory process. We have inhibited the arachidonic acid cascade with the nonsteroidal anti-inflammatory drugs, indomethacin (5 mg/kg/day) and phenylbutazone (10mg/kg/day). Forty-five New Zealand white rabbits were given a 0.25% cholesterol diet for 16 weeks in addition to the drugs or a placebo. Serial plasma lipid profiles were performed; aortic and peripheral arterial cholesterol content measurements were obtained at sacrifice. All groups showed markedly elevated plasma lipids. Thoracic aortic and carotid artery cholesterol contents were significantly decreased in both treated groups as compared to placebo (P < 0.05), despite the atherogenic lipid profiles. These results implicate the immune and inflammatory responses in the pathogenesis of atherogenesis. PMID- 8429642 TI - The role of the adrenals in the acute phase response to interleukin-1 and tumor necrosis factor-alpha. AB - The purpose of this study was to investigate the role of the adrenals, particularly the glucocorticoids, in the acute phase response following daily injections for 5 days of recombinant human interleukin-1 alpha,beta and tumor necrosis factor-alpha (TNF alpha). Adult weight-stable freely fed or pair-fed (to cytokine-injected mice) mice (C57Bl/6J) with and without adrenals were used. Adrenalectomized animals showed a sensitivity to both IL-1 alpha and -1 beta (40 ng IL-1/day) greater than 10-fold higher than that of normal mice (420 ng IL 1/day) in regard to mortality and anorexia. Microscopic examination of tissue specimens from adrenalectomized IL-1 alpha,beta-injected mice did not reveal any histologic alterations in lung, kidney, liver, brain, or gastrointestinal tract to explain the mortality. This mortality was not prevented by physiologic replacement doses of hydrocortisone (10-20 micrograms/day); however, a pharmacological dose of 2.5 mg hydrocortisone/day abolished completely the increased toxicity to IL-1 alpha,beta and the anorectic response to IL-1 alpha,beta and TNF alpha. Increased toxicity (mortality) was not observed in adrenalectomized animals with TNF alpha at the dose interval used (450 ng TNF alpha/day and lower). The hepatic acute phase response (liver weight and RNA and liver protein content) was increased by both IL-1 alpha,beta and TNF alpha in a glucocorticoid-independent way. The cytokine-induced alterations of plasma concentrations of acute phase proteins (serum amyloid P, transferrin, complement C3) were significantly dependent on glucocorticoids, while the decline in plasma albumin was not.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429643 TI - Interferon-gamma: a key immunoregulatory lymphokine. AB - Interferon-gamma is an important component of numerous host responses to inflammatory insults. Data from in vitro and animal studies suggest that this agent may have clinical uses in immunocompromised patients who are at increased risk for infectious complications. In addition to reviewing our current understanding of the in vitro and in vivo actions of interferon-gamma, this article cautions that the potential for adverse reactions exists when interferon gamma is administered to an immunologically competent host. PMID- 8429644 TI - Fulfilling expectations. PMID- 8429645 TI - Normocalcemic blood or crystalloid cardioplegia provides better neonatal myocardial protection than does low-calcium cardioplegia. AB - Although standard blood cardioplegia provides good myocardial protection for cardiac operations in adults, protection of the cyanotic, immature myocardium remains suboptimal. Calcium, which has been implicated in reperfusion injury and in the development of "stone heart" in mature myocardium, is routinely lowered in standard cardioplegic solutions. Immature, neonatal myocardium has lower intracellular calcium stores and is more reliant on extracellular calcium for contraction. To determine if normocalcemic cardioplegia would result in improved cardiac function in the neonatal heart, we conducted a series of experiments using an isolated, blood-perfused working heart model. Thirty-two neonatal piglet hearts (24 to 48 hours) were excised without intervening ischemia and were placed directly on a blood-perfused circuit. Baseline stroke work index was assessed. Hearts were then arrested with cold cardioplegic solution delivered at 45 mm Hg for 2 minutes: group I, low-calcium blood cardioplegic solution (Ca = 0.6 mmol/L); group II, normal-calcium blood cardioplegic solution (Ca = 1.1 mmol/L); group III, University of Wisconsin solution; and group IV, University of Wisconsin solution with added calcium (Ca = 1.0 mmol/L). Cardioplegic solution was administered every 20 minutes for 2 hours and topical hypothermia was used. Hearts were then reperfused with warm whole blood. Functional recovery, expressed as a percentage of control stroke work index, was determined minutes after reperfusion. Hearts preserved with normocalcemic cardioplegic solution (groups II and IV) had complete functional recovery at 60 minutes, whereas hearts preserved with low-calcium cardioplegic solution (groups I and III) achieved functional recoveries of only 80% and 65%, respectively, at a left atrial pressure of 9 mm Hg. Electron micrographs taken 1 hour after reperfusion showed minimal edema and only mild myofibrillar changes. They were identical in both the low-calcium and normocalcemic groups. Complete functional recovery is possible in immature myocardium when calcium is added to either blood or an intracellular crystalloid cardioplegic solution. The addition of calcium does not result in ultrastructural damage and does result in good functional recovery. PMID- 8429646 TI - Effect of calcium and preischemic hypothermia on recovery of myocardial function after cardioplegic ischemia in neonatal lambs. AB - Hypothermia has been reported to increase intracellular ionized calcium, which may aggravate injury resulting from ischemia and reperfusion. The effects of plasma ionized calcium concentration ([Ca2+]) during hypothermic perfusion on recovery after 2 hours of cold cardioplegic ischemia were evaluated in 32 isolated, blood-perfused neonatal lamb hearts. Three groups of hearts (B, C, and D) were perfusion-cooled for 10 minutes to a myocardial temperature of 17 degrees C and then arrested with St. Thomas' Hospital cardioplegic solution. Group A had 10 minutes of normothermic perfusion before cardioplegia. Group B had cooling with normal [Ca2+]. Group C had citrate added as cooling was started to lower [Ca2+] (0.26 mmol/L), and it was not normalized until 15 minutes into reperfusion. Group D received citrate plus Ca2+ to give normal [Ca2+] during cooling. Groups B and D showed a significantly reduced recovery (p < 0.05) in left ventricular systolic function (developed pressure and the rate of pressure rise) and diastolic function (stiffness constant) than groups A and C. During preischemic cooling, oxygen consumption per beat and coronary vascular resistance increased significantly in groups B and D, but both oxygen consumption and coronary vascular resistance were significantly lower in group C than in groups B and D so long as [Ca2+] was low. The data show that preischemic hypothermia results in reduced postischemic recovery of function compared with simultaneous induction of cardioplegia and hypothermia. Low [Ca2+] during preischemic hypothermia and early reperfusion offsets this deleterious effect of hypothermia. PMID- 8429647 TI - Myocardial distribution of cardioplegic solution after retrograde delivery in patients undergoing cardiac surgical procedures. AB - The myocardial distribution of both antegrade and retrograde cardioplegia for cardiac surgical intervention, after induction of cardioplegia via the aortic root, was directly assessed and compared in 19 patients by means of contrast echocardiography. Two-dimensional transesophageal echocardiographic images of the short axis of the left ventricle at the level of the papillary muscles were obtained after sonicated Renografin-76 microbubbles were injected into an aortic root and/or transatrial coronary sinus catheter during delivery of cardioplegic solution. Segmental distribution of cardioplegic solution was immediately noted in the myocardium at the time of contrast injections. In 11 of 18 patients (61%) cardioplegic solution was dispersed to all left ventricular myocardial segments after antegrade delivery. In 17 of 19 patients (90%) retrogradely delivered cardioplegic solution (after antegrade induction of cardioplegia in 18 of the 19 patients) was dispersed to all the left ventricular myocardial segments, including the septum. In 2 of the patients, initial lack of retrograde distribution of cardioplegic solution was remedied when the coronary sinus catheter was repositioned and contrast cardioplegic solution was reinjected. Imaging of the right ventricle was possible in only 4 of the 19 patients and revealed that after retrograde delivery, cardioplegic solution had been at least partially distributed to the right ventricle as well. We performed off-line videodensitometric analysis in 9 patients after retrograde delivery of cardioplegic solution. Mean peak pixel-intensity ratio of flow from the endocardium to the epicardium in the left ventricular free wall was 1.46 +/- 0.27, and mean peak pixel-intensity ratio of flow from the left to the right intraventricular septal endocardium was 1.39 +/- 0.33 (p < or = 0.05). PMID- 8429648 TI - Redo cardiac surgery: late bleeding complications from topical thrombin-induced factor V deficiency. AB - Bovine thrombin-induced factor V deficiency was though to be a very rare acquired coagulopathy, with only three documented cases. We report a series of nine patients seen during a period of 32 months; these patients had normal preoperative coagulation profiles, and this unique coagulopathy developed 1 to 2 weeks after cardiovascular operations. The coagulopathy was characterized by a markedly elevated prothrombin time (40.9 +/- 5.8 seconds), an elevated activated partial thromboplastin time (96.3 +/- 12.2 seconds), a study positive for lupus anticoagulation (9/9), and markedly decreased levels of factor V (0.09 +/- 0.03 U/ml) and factor XI (0.04 +/- 0.02 U/ml), respectively. All patients had been exposed to commercially available bovine thrombin during prior cardiovascular or vascular operations and received a second bovine thrombin challenge during the latest procedure. Coagulopathic bleeding developed in four of the nine patients. Bleeding was unrelated to absolute fall in factor V level, but cessation of hemorrhage appeared to correlate with improvement in factor V level. Treatment with vitamin K, fresh frozen plasma, and platelet infusion were all unsuccessful in altering prothrombin time or factor V levels. Intravenous gamma globulin was used in three patients, two of whom were bleeding. All three patients showed a transient increase in factor V levels. Bleeding stopped in one of the two patients; the other continued to bleed and subsequently died. The third patient was treated prophylactically to increase factor V levels in preparation for flap reconstruction of his sternum. His factor V level increased from 0.26 to 0.49 U/ml, and he underwent the procedure without incident. Bovine thrombin-induced factor V deficiency may have been previously unrecognized. This deficiency should be suspected in patients who have undergone redo cardiovascular operations and in whom marked elevations in their prothrombin time occur 7 to 10 days after exposure to bovine thrombin. The resulting coagulopathy, although usually self limited, has the potential to produce devastating bleeding complications. Intravenous gamma globulin (1 gm/kg during each of 2 days) has been used to increase factor V levels transiently but its role in therapy of this coagulopathy requires further investigation. PMID- 8429649 TI - Differential lung ventilation. Applications beyond the operating room. AB - Mechanical ventilatory support in the setting of unilateral lung disease offers unique problems in management. When the difference in airway resistance or lung compliance between the two lungs is exaggerated, conventional mechanical ventilation might lead to preferential ventilation with hyperexpansion of one lung and gradual collapse of the other. Differential ventilation has been advocated to avert this problem. We illustrate the use of this technique in the management of two patients with different underlying pathologic conditions. PMID- 8429650 TI - Interleukin-8 release and neutrophil degranulation after pediatric cardiopulmonary bypass. AB - Capillary leak after cardiopulmonary bypass operations for correction of congenital heart defects is universally seen in children and often causes significant morbidity and mortality. Since neutrophil-mediated endothelial injury has been implicated as a pathogenetic mechanism, a prospective controlled descriptive study was performed to investigate possible activation pathways during and after the bypass procedure. Eighteen children undergoing operations, nine with cardiopulmonary bypass and nine neurosurgical craniotomy (i.e., operations without bypass), had samples of arterial blood collected at intervals before, during, and after operations. In six of nine cardiac patients circulating interleukin-8 concentrations rose from less than 30 pg/ml to very high concentrations (> 500 pg/ml); in the remaining three patients small rises (peak 57 to 81 pg/ml) were also seen. In all nine, the rise commenced at the time of rewarming, toward the end of bypass, and peaked 1 to 3 hours thereafter. Interleukin-8 release correlated significantly with length of bypass. Interleukin 1 alpha and interleukin-1 beta were not found, and traces of tumor necrosis factor-alpha were detected in one patient only. Circulating elastase alpha 1 antitrypsin concentrations rose simultaneously and correlated significantly with interleukin-8 (p < 0.001) in patients with cardiac disease, as did absolute neutrophil counts (p < 0.001). In contrast, only one of nine patients with neurosurgical disease (undergoing an unusually long operation and exchange transfusion) had a rise in circulating interleukin-8 to levels greater than 500 pg/ml (p < 0.01). The two samples from this patient with elevated interleukin-8 were the only neurosurgical samples with elevated elastase. This study demonstrates the release of interleukin-8 into the circulation after pediatric hypothermic cardiopulmonary bypass and supports the suggestion that this cytokine plays a role in the pathophysiology of capillary leak through neutrophil degranulation. PMID- 8429651 TI - Allograft replacement of the trachea. Experimental synchronous revascularization of composite thyrotracheal transplant. AB - We describe a method for experimental tracheal transplantation that uses synchronous revascularization based on the blood supply of the related thyroid gland. This study compares the structural integrity and histologic features of nonvascularized tracheal transplants, revascularized thyrotracheal composite allografts, with and without immunosuppression. Eighteen adult beagle dogs underwent tracheal transplant operations with one of the preceding methods. Postmortem specimens were studied macroscopically and histologically, 3 to 28 days postoperatively. Proximal, mid, and distal segments of each graft (twelve ring segments) were examined microscopically. Nonvascularized tracheal transplants necrosed completely as early as 3 days postoperatively. The vascularized composite thyrotracheal allografts survived for up to 28 days. Six dogs did not receive immunosuppressive agents; tracheal cartilage was preserved in this group, but soft tissue necrosis developed. Six dogs received cyclosporine; tracheal cartilages and all soft tissues remained histologically intact. This difference may be due to early tracheal cartilage ischemia in the nonvascularized group, rejection of the soft tissues in the nonimmunosuppressed group, and viability of all structures in the vascularized and immunosuppressed group. Revascularization of the transplanted trachea with the thyroid arteries maintains the vascularity and hence the viability of the trachea. A reliable substitute for long-segment tracheal resections is finally found. We predict that clinical application of this technique will solve a major problem in the surgery of the airways. PMID- 8429652 TI - Intrathoracic stomach. Presentation and results of operation. AB - Between January 1, 1980, and December 31, 1990, 147 patients (93 female and 54 male) were found to have an intrathoracic stomach. Median age was 69 years (range 34 to 89). Signs and symptoms occurred in 140 patients (95.2%) and were primarily obstructive. They included postprandial pain in 87 (59.2%), vomiting in 46 (31.3%), and dysphagia in 44 (29.9%); only 23 patients (15.7%) had symptoms of gastroesophageal reflux. Anemia was present in 31 patients (21.1%) and melena in 3. Elective repair was done in 119 patients and included an uncut Collis-Nissen repair in 81 patients (68.1%), a Belsey Mark IV repair in 19 (16.0%), a Nissen repair in 17 (14.3%), and a Harrington (anatomic) repair in 2 (1.7%). Thirty-two patients had complications (26.9%). There were no operative deaths. Median follow up was 42 months. Results were excellent in 69 patients (60.0%), good in 38 (33.0%), fair in 6 (5.2%), and poor in 2 (1.7%). Five patients had emergency operations for suspected strangulation; three had gastric necrosis, and one died. Two of the four operative survivors had excellent results. Twenty-three other patients were followed up with medical management for a median of 78 months (range 12 to 268 months). In four patients progressive symptoms developed, and one patient died from aspiration. We conclude that patients with an intrathoracic upside-down stomach who have obstructive symptoms at initial presentation should undergo repair and that elective operation is safe and effective. Gastric strangulation, however, is rare. PMID- 8429653 TI - Thoracic esophageal diverticula. Why is operation necessary? AB - Diverticula of the thoracic esophagus are uncommon disorders. The indications for surgical intervention in asymptomatic or minimally symptomatic patients are unclear. Among 20 patients referred during a 20-year period, 6 were male and 14 female, with a median age of 65 years. Two had had previous diverticulectomies. Dysphagia was present in 9 (45%) and regurgitation in 11 (55%). Nine patients had severe nocturnal cough with symptoms of aspiration. In two of these nine and in three other patients (25%), pulmonary symptoms were the only manifestation of disease, with no or minimal esophageal symptoms. In one patient the diagnosis of the presence of bronchial asthma for several years was incorrect; one patient had massive aspiration before hernia repair, in one a bronchoesophageal fistula and lung abscess developed, and two had severe persistent cough. All patients had a diagnostic barium esophagogram and endoscopy. Operation was performed in 17 patients, whereas three others declined operation. There was one hospital death. Follow-up is complete on 17 of 19 patients until June 1991. All operative survivors but one are free of symptoms. Of three patients refusing operation, one died of aspiration pneumonia, another died of myocardial infarction, and one with severe dysphagia is living. Because of the prevalence of aspiration (45%) and the potential for life-threatening pulmonary complications in some patients (15%), we conclude that operative intervention should be undertaken in all patients with thoracic esophageal diverticula regardless of the presence or absence of symptoms. PMID- 8429654 TI - Transhiatal esophagectomy for benign and malignant disease. AB - Transhiatal esophagectomy has been performed in 583 patients with diseases of the intrathoracic esophagus: 166 (28%) benign and 417 (72%) malignant (6% upper, 28% middle, and 66% lower third and cardia). The benign esophageal diseases included strictures (40%); neuromotor dysfunction-achalasia (24%), esophageal spasm (8%); recurrent gastroesophageal reflux (16%); acute perforation (5%); acute caustic injury (2%); and others (3%). Among the patients with benign disease, 60% had undergone at least one prior esophageal operation. Transhiatal esophagectomy was possible in 97% of patients in whom it was attempted, 19 patients (13 with benign disease and 6 with carcinoma) requiring addition of a thoracotomy for esophageal resection. Esophageal resection and reconstruction were performed in a single operation in all but 5 patients. The esophageal substitute was positioned in the posterior mediastinum in the original esophageal bed in 96%. Stomach was used to replace the esophagus in 553 patients (95%) and colon in 28 (5%) who had undergone prior gastric resections. Overall hospital mortality was 5% in patients with benign disease and 5% in those with carcinoma. There was 1 intraoperative death caused by uncontrollable hemorrhage. Complications included intraoperative entry into a pleural cavity necessitating a chest tube (74%), anastomotic leak (9%), recurrent laryngeal nerve paralysis (3%), and chylothorax and tracheal laceration (< 1% each). Three patients required reoperation for mediastinal bleeding. Average intraoperative blood loss was 875 ml (1023 ml for benign disease and 817 ml for carcinoma). Of the surviving patients, 88% were discharged able to swallow within 3 weeks of operation and 78% within 2 weeks. The actuarial survival of the patients with carcinoma is similar to that reported after more traditional transthoracic esophagectomy. Among patients with benign disease, good or excellent functional results have been achieved in nearly 70% after a cervical esophagogastric anastomosis. Although approximately 44% have required one or more anastomotic dilations within 1 to 3 months of operation, true anastomotic strictures have developed in 10%. Clinically troublesome nocturnal reflux has occurred in 3%. Transhiatal esophagectomy is feasible in most patients requiring esophageal resection for either benign or malignant disease and is a safe, well tolerated operation if performed with care and for the proper indications. PMID- 8429655 TI - A new video-assisted thoracoscopic surgical technique for interruption of patient ductus arteriosus in infants and children. AB - Classic surgical interruption of patent ductus arteriosus was partially replaced by transcatheter endovascular closure in 1971. We describe a new technique for ductus closure by video-assisted thoracoscopic surgical interruption, derived from video-assisted endoscopic surgery. With the patient under general anesthesia and intubated, two 5 mm holes were made through the left thoracic wall. A video camera and specially devised surgical tools were introduced; such as scissors, dissectors, and clip appliers. The ductus was dissected, and two titanium clips were applied, completely interrupting the ductus. Thirty-eight patients were operated on from April 1991 to April 1992. Mean age was 23.3 months (range 1.5 to 90 months) and mean weight was 9.5 kg (range 2.4 to 25 kg). Six had associated lesions not necessitating immediate surgical treatment. All had successful closure of the patent ductus with the video-assisted technique, but two needed two such procedures, one because of incomplete immediate ductus closure and one because of partial opening of the clip after 24 hours. One patient had recurrent laryngeal nerve injury and four had pneumothorax on the left side. The usual hospital stay was 2 or 3 days. There were no other complications and no deaths. Video-assisted thoracoscopic surgical interruption was a rapid, safe, and successful technique for closure of the patent ductus arteriosus. Better dissection of the ductus decreased the risk of recurrent laryngeal nerve injury and that of clip opening. In the last 26 patients, in whom a 2 mm multiperforated catheter was used for chest drainage during the first hours, no pneumothoraces occurred. Video-assisted thoracoscopic interruption of the patent ductus arteriosus is feasible in low-weight infants, whereas transcatheter endovascular closure of the ductus usually is not possible. The technique will be applied to premature infants with new instruments designed for the size of these patients. PMID- 8429656 TI - Results of surgical repair of congenital supravalvular aortic stenosis. AB - Our experience with congenital supravalvular aortic stenosis dates from 1977 to 1991. Thirteen patients, aged 2 days to 38 years (mean 7.6 years, median 3.8 years), had surgical repair of the lesion. The preoperative peak-to-peak systolic gradients ranged from 25 to 110 mm Hg (mean 64 mm Hg). Four patients had trace to mild aortic insufficiency. Surgical repair was accomplished by several techniques that have evolved over time. There was one death in a 2-day-old neonate who also had severe hypertrophic cardiomyopathy. The 12 survivors had postoperative gradients of 0 to 30 mm Hg (mean 10 mm Hg) in the supravalvular region. The mean reduction in gradient was 48 mm Hg. A new technique employing all autologous aortic tissue is described. PMID- 8429657 TI - The management of severe subaortic stenosis, ventricular septal defect, and aortic arch obstruction in the neonate. AB - Neonates with ventricular septal defect and aortic arch obstruction frequently have subaortic stenosis resulting from posterior deviation of the infundibular septum. Because the aortic anulus is often hypoplastic, making direct resection of the infundibular septum through the standard transaortic approach difficult, the optimal method of repair is uncertain. From September 1989 through November 1991, seven patients with ventricular septal defect, coarctation (n = 4), or interrupted aortic arch (n = 3) and severe subaortic stenosis underwent repair with use of a technique that included transatrial resection of the infundibular septum. Their ages ranged from 5 to 63 days (median 15 days) and weights from 1.3 to 5.4 kg (mean 3.1 kg). Only one patient was older than 1 month. The systolic and diastolic ratios of the diameter of the left ventricular outflow tract to that of the descending aorta were 0.53 +/- 0.09 mm (standard deviation) and 0.73 +/- 0.11, respectively. At operation, the posteriorly displaced infundibular septum was partially removed through a right atrial approach by resecting the superior margin of the ventricular septal defect up to the aortic anulus. The resulting enlarged ventricular septal defect was then closed with a patch to widen the subaortic area. In each patient the aortic arch was repaired by direct anastomosis. All patients survived operation; there was one late death from noncardiac causes 3 months after repair. The survivors remain well from 3 to 14 months after repair (mean 8 months). All are in sinus rhythm and none has a residual ventricular septal defect. One patient underwent successful balloon dilation of a residual aortic arch gradient late after repair. No patient has significant residual subaortic stenosis, although one has valvular aortic stenosis. This series suggests that in neonates with ventricular septal defect and severe subaortic stenosis resulting from posterior deviation of the infundibular septum, direct relief can be satisfactorily accomplished from a right atrial approach. This method provides effective widening of the left ventricular outflow tract and is superior to palliative techniques or conduit procedures. PMID- 8429658 TI - Successful thrombectomy for thrombosis of the right side of the heart after the Fontan operation. Report of two cases and review of the literature. AB - We performed operative thrombectomy for the treatment of cardiac thrombosis on the right side of the heart in two patients; thrombosis occurred after a total cavopulmonary connection (modified Fontan operation). In the first case, thrombosis occurred approximately 6 months postoperatively, and the thrombus was evacuated without cardiopulmonary bypass. In the second case, an emergency open atrial thrombectomy and revision of the stenotic inferior cavopulmonary anastomosis was performed on the tenth postoperative day. Review of the literature identified 12 previously reported cases that were similar. In these 12 cases 6 patients died, 5 of them early after diagnosis and institution of treatment. We believe that appropriate management should include thrombectomy and revision if a surgically remediable cause of the thrombosis is identified; otherwise, thrombolytic therapy should be initiated. The use of heparin and then sodium warfarin (Coumadin) has also been successful. Risk of thrombosis of the right side of the heart after the Fontan repair may be minimized by the use of prophylactic anticoagulation in high-risk patients soon after the Fontan operation. PMID- 8429659 TI - Late results of valve replacement with the Bjork-Shiley valve (1973 to 1982) AB - Cardiac valve replacement with use of only the Bjork-Shiley prosthesis was performed in 1253 patients between January 1973 and December 1982. There were 828 patients having aortic valve replacement, 280 patients having mitral valve replacement, and 145 patients having double valve replacement with aortic and mitral valve prostheses. Patient outcome was stratified according to multiple variables, including valve position and valve model (spherical versus convexo concave discs). No valve failure due to strut fracture was identified in 26 high risk patients (mitral valve replacement with greater than or equal to 29 mm implanted in patients less than or equal to 50 years of age) followed up for a mean of 10 years postoperatively. Fifteen patients had late thrombosis of their Bjork-Shiley prosthesis (0.28 per 100 patient-years), but there was no significant difference in risk of valve thrombosis comparing the spherical and convexo-concave discs (0.27 per 100 patient-years versus 0.27 per 100 patient years). One hundred two patients had 128 thromboembolic episodes; rates of thromboembolism after aortic valve replacement, mitral valve replacement, and double valve replacement were 2.1, 4.3, and 4.6 per 100 patient-years, respectively. Percentages of patients free from thromboemboli after aortic valve replacement, mitral valve replacement, and double valve replacement were 93% +/- 1%, 86% +/- 2%, and 89% +/- 3% at 5 years postoperatively and 87% +/- 2%, 79% +/- 5%, and 77% +/- 8% 10 years postoperatively. There was no significant difference in the rates of thromboemboli for spherical and convexo-concave discs for all patients and for each of the subgroups. Ten-year actuarial survival estimates for patients dismissed alive from the hospital after aortic valve replacement, mitral valve replacement, and double valve replacement with the Bjork-Shiley valve were 65% +/- 4%, 63% +/- 5%, and 55% +/- 8%, respectively. Overall event-free survival (freedom from death, thromboembolism, anticoagulant-related bleeding, endocarditis, and reoperation) was similar for the three patient groups. Performance of the Bjork-Shiley valve as judged by late patient follow-up is similar to other mechanical valves, and modifications in disc design do not appear to have reduced the threat of late valve thrombosis and thromboemboli. Evidence does not support elective explantation of this prosthesis. PMID- 8429660 TI - Sound spectral analysis of prosthetic valvular clicks for diagnosis of thrombosed Bjork-Shiley tilting standard disc valve prostheses. AB - To evaluate clinical usefulness of sound spectral analysis in the early detection of prosthetic thrombosis, we analyzed specific patterns of thrombus formation in Bjork-Shiley tilting standard disc prostheses in relation to the sound spectral analysis of their click sounds. Among a total of 365 Bjork-Shiley tilting standard disc prostheses, nine became thrombotic. These consisted of seven prostheses in the mitral position and two in the aortic position, and the affected valves were replaced in all cases. The sound spectral analysis system includes a wide-frequency microphone and a sound spectral analyzer. To quantify the amplitude of the metallic clicks that are specific to the prosthetic valve sounds, we used a normalized maximal frequency as a diagnostic parameter for valve thrombosis. Thrombus formation was localized at the minor strut in two cases, and these demonstrated abnormally low normalized maximal frequency values only on opening clicks. In four cases thrombus formations were noted at both minor and major struts, and these showed abnormally low normalized maximal frequency values on both opening and closing clicks. In two cases with pannus formation, no abnormalities were found by sound spectral analysis. In one case, which had demonstrated abnormally decreased normalized maximal frequency values of both opening and closing clicks, the normalized maximal frequency values were normalized after thrombolytic therapy, suggesting resolved thrombosis. Reoperation, which was performed for concomitant complication of perivalvular leak, revealed no noticeable thrombus formation. The thrombi were seen on the minor strut during surgical procedures in all cases in which valve thrombosis was indicated by the sound spectral analysis. Therefore, the minor strut is considered to be the prevalent area of thrombus formation, and diagnosis could be made after observation of abnormally low normalized maximal frequency on opening clicks. We found the sound spectral analysis to be an extremely useful diagnostic tool for early detection of thrombosis in the Bjork-Shiley tilting standard disc prostheses, especially because the opening clicks of the tilting disc valve were too low in amplitude for auscultation to detect the existence of mild abnormality. PMID- 8429661 TI - In situ repair of mycotic aneurysm of the ascending aorta. AB - Between 1969 and 1990 six patients (aged 14 to 64 years, mean 43 years) underwent in situ reconstruction for mycotic aneurysm of the ascending aorta. The primary source of infection was endocarditis in three patients (subacute bacterial endocarditis [n = one patient], sepsis with acute endocarditis [n = one patient]), sepsis with sternal osteomyelitis in one, sepsis with purulent pericarditis in one, and generalized febrile illness in one. In five of six patients the treatment consisted of the excision of changed tissue combined with a composite graft (n = one patient), a xenopericardial patch repair (n = one patient), a Dacron graft repair and aortic valve replacement (n = one patient), a Dacron graft repair alone (n = one patient), and a lateral suture combined with double valve replacement (n = one patient). In one patient with perforation of the mycotic aneurysm into the pulmonary artery, the place of rupture was oversewn without excision of the aortic or pulmonary artery tissue. Two patients with local pericardial inflammation were reoperated on during the hospital stay; one of them because of recurrent mycotic aneurysm of the ascending aorta at the other location and the other because of infection of the suture line after the Dacron patch repair. Antibiotic therapy was intravenously administered for 2 to 12 weeks postoperatively and continued orally for 4 to 8 weeks. The mean observation time was 6 years (range 4 months to 16 years). There was no late graft infection, except the chronic infection of the suture line in one patient who died suddenly 4 months after the operation. There was no early death, and there were three late deaths (chronic myocardial failure, one patient, chronic renal failure, one patient, sudden death, one patient). We concluded that in situ reconstruction for mycotic aneurysm of the ascending aorta combined with prolonged antibiotic therapy is an appropriate procedure with satisfactory early and good long-term results. PMID- 8429662 TI - Extensive cryoablation of the left ventricular posterior papillary muscle and subjacent ventricular wall. Impact on mitral valve function and hemodynamics. AB - Ventricular tachycardias that originate from the inferior left ventricular wall may necessitate transmural ablation involving the posterior papillary muscle of the mitral valve. The effect on mitral valve function and hemodynamics of extensive cryoablation of the left ventricular posterior papillary muscle and subjacent ventricular wall was studied in 16 dogs. Two sham experiments were done. All dogs were studied preoperatively and postoperatively by pulsed Doppler and two-dimensional echocardiography. Left ventricular angiographic and hemodynamic studies were performed preoperatively in six treated dogs and two control dogs and in all dogs at the end of follow-up (1, 3, or 6 months). Postmortem studies were performed in all dogs. The cumulative probability of freedom from mitral regurgitation at 2 months was 0.43 +/- 0.14. Thereafter no new cases of mitral regurgitation could be demonstrated. The angiographic degree of mitral regurgitation was mild in five and moderate in two dogs and did not increase from 3 to 6 months. One dog with acute severe mitral regurgitation died early of heart failure. A significant increase in left ventricular end-diastolic and mean pulmonary capillary wedge pressure of 9.4 +/- 2.5 mm Hg and 6.4 +/- 2.6 mm Hg, respectively, was found in treated dogs at 3 months. These results suggest that extensive cryoablation of the left ventricular posterior papillary muscle and subjacent ventricular wall can be accomplished with an acceptable risk of mild to moderate mitral regurgitation, and without serious detrimental effect on left ventricular function. Retraction is probably the main mechanism of mitral regurgitation. PMID- 8429663 TI - Physiologic evaluation of pulmonary function in the candidate for lung resection. AB - From July 1, 1974, to December 31, 1990, 2340 patients who underwent pulmonary resection were evaluated by comprehensive analysis of pulmonary function. Pulmonary function test criteria for resection were (1) pneumonectomy: forced expiratory volume in 1 second greater than 2 L; forced expiratory flow rate from 25% to 75% greater than 1.6 L; maximum voluntary ventilation greater than 55%; (2) lobectomy: forced expiratory volume in 1 second greater than 1 L; forced expiratory flow rate from 25% to 75% greater than 0.6 L; maximum voluntary ventilation greater than 40%; (3) wedge or segmental resection: forced expiratory volume in 1 second greater than 0.6 L; forced expiratory flow rate from 25% to 75% greater than 0.6 L; maximum voluntary ventilation greater than 35%. Split perfusion lung scan and Reichel exercise stress testing were utilized as indicated. When these values of pulmonary function have been applied, a more precise method of selecting patients for various types of pulmonary resection has resulted in a lower mortality while denying operation to less than 1% of the patients who are considered for surgical resection. PMID- 8429664 TI - Improved recovery of heart transplants with a specific kit of preservation solutions. AB - In the course of cardiac transplantation, donor hearts undergo a four-step sequence of events (arrest, cold storage, global ischemia during implantation, and reperfusion) during which myocardial damage can occur. We tested the hypothesis that the functional recovery of these hearts could be improved by exposure to two interdependently formulated preservation solutions throughout this four-step sequence. Solution I was used as a perfusion and storage medium during the first three steps, and solution II served as a modified reperfusate. The two solutions share the following principles of formulation: prevention of cell swelling (high concentrations of mannitol, a myocardium-specific impermeant) calcium overload (ionic manipulations), and oxidative damage (reduced glutathione) and enhancement of anaerobic energy production (glutamate). The two solutions differ with respect to the calcium content and buffering capacity. One hundred rat hearts perfused with isolated isovolumic buffer were subjected to cardioplegic arrest; cold (2 degrees C) storage for 5 hours, global ischemia at 15 degrees C for 1 hour, and normothermic reperfusion for 1 additional hour. In a first series of experiments (70 hearts), our kit of solutions was compared with six clinical preservation regimens that involved cardiac arrest with St. Thomas' Hospital or University of Wisconsin solutions followed by storage of the hearts in saline, Euro-Collins, St. Thomas' Hospital, or University of Wisconsin solutions. In a second series of experiments (30 hearts), the effects of the kit were more specifically investigated in relation to two types of additive--oncotic agents (dextran) and thiol-based antioxidants (reduced glutathione and N-acetyl-L cysteine). According to comparisons of maximal rate of ventricular pressure increase and left ventricular compliance after reperfusion, the best myocardial protection was afforded by our kit of solutions. The addition of dextran during storage did not provide additional protection. Conversely, the omission of reduced glutathione was clearly detrimental; the replacement of reduced glutathione with N-acetyl-L-cysteine failed to improve recovery beyond that provided by antioxidant-free solutions, thereby suggesting the importance, in this model, of an anti-free radical compound that, like reduced glutathione, is operative extracellularly. We conclude that the preservation of heart transplants can be improved with the sequential use of two closely interrelated solutions, the formulations of which integrate the basic principles of organ preservation with those of myocardium-specific metabolism. PMID- 8429665 TI - Preliminary experimental results of a new resorbable biomaterial as pericardial substitute. PMID- 8429666 TI - Invited letter concerning: critical aortic stenosis. PMID- 8429667 TI - Total anomalous pulmonary venous drainage by double connection corrected by ascending vein and coronary sinus repair. PMID- 8429668 TI - Patterns of ductal tissue in coarctation of the aorta in early infancy. PMID- 8429669 TI - Transposition of the great arteries with posterior aorta. PMID- 8429670 TI - Aneurysm of the membranous ventricular septum in transposition of the great arteries. PMID- 8429671 TI - The hemostatic effect of autologous platelet-rich plasma versus autologous whole blood after cardiac operations: is platelet separation really necessary? PMID- 8429672 TI - The reoxygenation phenomenon. PMID- 8429673 TI - Invited letter concerning: cardiac stun syndrome. PMID- 8429674 TI - Differential effects of bryostatin 1 on human non-Hodgkin's B-lymphoma cell lines. AB - Bryostatin 1 (Bryo1), a macrocyclic lactone and a protein kinase C activator, is extracted and purified from the marine bryozoan Bugula neritina. In this study we describe its effect on morphology, surface immunophenotype, acid phosphatase (AcP), tartrate-resistant acid phosphatase (TRAP), proliferation and cell cycle of non-Hodgkin's B-lymphoma cell lines representing four differentiation stages. Except for the WSU-BL, a high-grade SCNCL, all other cell lines showed obvious changes in their morphology when treated with 200 nM Bryo1. Phenotypically, a dramatic decrease of CD10 and induction of CD11c and BL7 on some cell lines consistent with further B-cell differentiation was seen. The lines in control cultures showed variable expression of AcP and TRAP. Following treatment with Bryo1, there was a general increase in AcP expression except in WSU-BL line. WSU FSCCL and WSU-DLCL were TRAP-negative but became TRAP-positive when treated with Bryo1. Cell growth and cycle analysis during treatment of different cell lines revealed evidence of strong, moderate, or no growth inhibition by Bryo1 compared with control cultures. Our results indicate that Bryo1 shows differentiation effects on low-grade FSCCL, intermediate-grade FLCL and high-grade DLCL, and stimulatory or no effect on high-grade SCNCL. Since Bryo1 does not have tumor promoting activity, it has a potential therapeutic role as a B-cell differentiating agent. PMID- 8429675 TI - Interleukin-2 receptor alpha chain on acute myelocytic leukemia cells is involved in cell-to-cell interactions. AB - Leukemic cells from 21 to 197 adult patients with de novo acute myelocytic leukemia (AML) were positive for IL-2R alpha chain (IL-2R alpha), whereas IL-2R beta chain (IL-2R beta), which is responsible for IL-2 signal transduction, was not found on leukemic cells from any of these cases tested. The expression of IL 2R alpha was closely associated with that of adhesion molecules CD4, CD11b and CD22, and endopeptidase CD10. None of the IL-2R alpha (+) AML cells responded to recombinant human IL-2. These data suggest that IL-2R alpha on AML cells may not be involved in cellular proliferation as one of growth factor receptors but may have a role in the control of cell-to-cell interactions. PMID- 8429676 TI - Characterization of K562 cells following introduction of a mutant N-ras gene. AB - A mutant human N-ras gene (codon 61, C to A substitution) was electroporated into the human leukemic cell line K562, originally derived from a patient with chronic myeloid leukemia (CML) in blast crisis. Despite confirmation of mutant N-ras gene integration and expression, mutant transfected cells exhibited no growth advantage when characterized in suspension cultures and clonogenic assays, and serum deprivation impaired proliferation of both normal and mutant N-ras transfected cells equally. A subclone containing a mutant N-ras gene displayed a proliferation rate and differentiation potential identical to that of non transfected cells. The failure of N-ras mutations to modify K562 cell behavior is in keeping with the infrequent observation of N-ras mutations in blastic transformation of CML. PMID- 8429677 TI - Over expression of insulin-like growth factor receptor type-I in T-cell lines infected with human T-lymphotropic virus types-I and -II. AB - To determine if aberrant expression of tyrosine kinase growth factor receptors may be related to the cell transformation capabilities of human T-lymphotropic viruses (HTLVs), we examined the expression of the epidermal growth factor receptor (EGF-R), insulin receptor (INS-R), and insulin-like growth factor receptor type-I (IGFR-I) in cell lines infected with HTLV type I (MT-2, HuT-102) and HTLV type II (Mo-T). Levels of mRNA transcripts for IGFR-I were significantly higher in both MT-2, HuT-102 (HTLV-I) and Mo-T (HTLV-II) cell lines than in uninfected cell lines (HuT-78, Jurkat); no detectable levels of EGF-R or INS-R mRNA transcript were observed in HTLV-infected or uninfected cell lines. Southern blot analysis demonstrated that no amplification or rearrangement of the IGFR-I gene occurred in either the MT-2 or Mo-T cell line. Flow cytometry analysis demonstrated that while IGFR-I protein was constitutively expressed on the cell surface in both MT-2 and Mo-T cell lines, neither EGF-R nor INS-R proteins could be detected. Ligand binding studies with MT-2 and Mo-T cell lines demonstrating binding of 125I insulin-like growth factor type-1 (IGF-I) in a dose-dependent manner and this response could be inhibited by increasing concentrations of cold IGF-I. These data demonstrate that deregulated expression of functional IGFR-I, the regular component of the growth control machinery of normal cells, may contribute to cellular proliferation and eventual transformation in HTLV-I- and HTLV-II-infected cell lines. PMID- 8429678 TI - Changes of buoyant density during the S-phase of the cell cycle. Direct evidence demonstrated in acute myeloid leukemia by flowcytometry. AB - Studies with synchronized or exponentially growing bacteria and mammalian cell lines are not able to demonstrate small changes in buoyant density during the cell cycle. Flowcytometric analysis of density separated acute myeloid leukemia cells, a system not dependent on time-related variables, shows that the cellular buoyant density increases slightly with up to 0.008 g/ml during the S-phase, at least in cryo-preserved cells used in this study. This contrasts with the generally accepted belief that S-phase cells have a lower or constant buoyant density. A practical implication is that separation of cell (sub)populations based on differences in buoyant density could be flawed to the extent that these populations contain S-phase cells. PMID- 8429679 TI - Preleukemic proliferative changes in murine bone marrow after single and multiple 7,12-dimethylbenz(a)anthracene (DMBA)-applications. AB - The effect of a leukemia-inducing treatment on early changes in kinetic parameters of murine bone marrow cells were investigated. Mice were treated i.p. one, four and eight times at biweekly intervals with 1 mg DMBA. Up to nine weeks after the last injection, CFU-S number, proliferation ability of bone marrow cells (PF), cell doubling time (td) and the compartment ratio (CR) were measured. Following multiple DMBA injections, CFU-S number and PF were decreased whereas CR and td increased, thus indicating persisting stem cell injury and proliferative compensation in the hemopoietic amplification compartment. A single DMBA injection had no effect. It is concluded that a first DMBA injection induces cytotoxic (and genotoxic) damage in the bone marrow leading simultaneously to a strong proliferation stimulus and a hindered proliferation ability of HSC, some of which will be predisposed for further mutagenic treatment. The following DMBA injections meet strongly proliferating HSCs, thus enhancing the probability for the loss of proliferation control/terminal differentiation. PMID- 8429680 TI - Functional interleukin-2 receptor on a Tac negative human leukaemia T-cell line. AB - Cell line PER-423 was derived from the cells of a patient with an immature acute T-lymphoblastic leukaemia and the growth of this human cell line is strictly dependent on interleukin-2 (IL-2). PER-423 cells express the p75 (beta) subunit of the IL-2 receptor (IL-2R beta), while the p55 chain (IL-2R alpha) is not detectable by immunofluorescence. The analysis of the IL-2R revealed that it is of intermediate affinity and the median effective IL-2 concentration for PER-423 cells (EC50 value) was determined to be 1.44 +/- 0.29 nM. Chemical crosslinking studies showed that the receptor consists of one polypeptide of approximately 95 kDa as well as a doublet of 70 kDa and 60 kDa and does not include the IL-2R alpha-chain. The steady-state mRNA level for the p75 subunit was similar to that present in a cell line expressing an IL-2R alpha+ beta+, while only traces for the alpha-chain were detectable. PER-423 cells can be induced to express the alpha-chain of the IL-2R on the cell surface, concomitant with a much reduced EC50 level. Since cell line PER-423 is functionally dependent on IL-2, it provides an ideal model for IL-2 signal transduction studies and for investigations focusing on the requirements for ligand binding vs activation. PMID- 8429681 TI - Normal hematopoietic reconstitution following ASTA-Z 7557-purged grafts in the absence of in vitro CFU-GM colony growth. AB - Hematopoietic reconstitution was assessed in 26 consecutive patients who underwent autologous bone marrow transplantation (ABMT) with ASTA-Z 7557 purged bone marrows. Of the 26, 17 had acute non-lymphoblastic leukemia (ANLL), 7 had acute lymphoblastic leukemia (ALL), 1 had non-Hodgkin's lymphoma (NHL) and 1 had multiple myeloma (MM). Twelve patients had practically no CFU-GM growth after ASTA-Z treatment. No statistical difference was observed in hematopoietic reconstitution between patients with or without in vitro CFU-GM colony growth. No significant linear correlation was observed between CFU-GM content in the treated graft and day of engraftment. In vitro CFU-GM growth has no predictive value in assessing hematopoietic recovery in vivo. Patients with no CFU-GM growth demonstrated normal engraftment. PMID- 8429683 TI - Role of transforming growth factor-beta in chronic lymphocytic leukemia. AB - TGF-beta is an important immunoregulator as it suppresses proliferation and function of B- and T-lymphocytes. In the present study we have examined the cellular localization and secretion of TGF-beta in B-cells from normal donors and patients with CLL and have assessed the influence of TGF-beta 1 on DNA synthesis in these cells. Using anti-LC(1-30)--a polyclonal anti-TGF-beta 1 antibody--TGF beta was localized to discrete sites within the cytoplasm of both normal and malignant lymphocytes. These areas co-localized with areas detected by an antigranule antibody (D545), suggesting that TGF-beta may be stored within cytoplasmic secretory vesicles. Both normal B- and CLL cells contained low or undetectable levels of TGF-beta mRNA and secreted low and equivalent amounts of TGF-beta. Compared to untreated cells, DNA synthesis was reduced by TGF-beta 1 to a mean +/- S. E. of 0.84 +/- 0.07 in CLL cells and this was significantly less (p < 0.001) than that observed in normal B-cells (mean +/- S. E. of control, 0.12 +/ 0.02). In 3 of the 18 patients, TGF-beta 1 stimulated DNA synthesis. The reduced inhibition of leukemic cell DNA synthesis by TGF-beta 1 in CLL may provide these cells with a growth or survival advantage over normal lymphocytes and contribute to their selective accumulation. PMID- 8429682 TI - Biodistribution and in vivo antitumor efficacy of the systemically administered anti-human T-leukemia immunotoxins and potentiation of their efficacy by alpha interferon. AB - In this study, the systemically administered anti-human T-leukemia immunotoxins (ITs) are shown to be effective for tumor suppression in Ichikawa T-leukemia bearing nude mice. In addition, their antitumor efficacy was markedly potentiated by recombinant human IFN-alpha. The combination of ITs and IFN-alpha effectively killed the tumor in the majority of the treated mice; 9 of the 12 treated mice survived tumor-free for as long as they were followed, i.e. for 140 days. Two different ITs, SN1-ricin A chain (RA) and SN2-RA, were used together to minimize the problem of tumor heterogeneity; monoclonal antibodies SN1 and SN2 are directed toward two different human T-leukemia associated cell surface antigens. In the biodistribution experiments, the paired label technique was used to include a reliable internal control. In an experiment, equal amounts of 125I-SN1 RA and 131I-labelled isotype-matching control IgG (IgG1-kappa)-RA were mixed and administered i.v. into tumor-bearing nude mice. In a separate experiment, a mixture of equal amounts of 125I-SN2-RA and 131I-control IgG-RA was administered i.v. This technique allowed us to distinguish the immunospecific uptake from the non-immunospecific uptake of ITs into individual organs. The present results clearly show that both SN1-RA and SN2-RA are specifically localized in tumors after systemic administration. For instance, 24 h after the administration of a radiolabelled mixture, the ratio of 125I/131I in the tumor was 7.0 and 23.5, respectively, for the SN1-RA/control IgG-RA mixture and the SN2-RA/control IgG-RA mixture. Such high ratios of 125I/131I were detected in the tumors throughout the experiments between 30 min and 24 h after the administration of the paired label mixture. PMID- 8429684 TI - Mechanisms of allogeneic stimulation induced tumor necrosis factor-alpha (TNF alpha) production. AB - We investigated the mechanisms of allogeneic stimulation induced TNF-alpha production in vitro by using human peripheral blood mononuclear cells (PBMC) and Daudi lymphoblastoid B-cells. PBMC produced TNF-alpha in response to mitomycin C treated or paraformaldehyde-fixed Daudi cells, reaching a peak level after 4-6 h of culture. Monocytes were identified as the major source of TNF-alpha produced during allogeneic cell interaction. The second potent producer of TNF-alpha was E rosette non-forming natural killer cells. Purified T-cells did not produce significant levels of TNF-alpha, even in the presence of IL-1 and IL-6. Interleukin-4 (IL-4) down-regulated TNF-alpha production by monocytes, but in contrast interferon-gamma (IFN-gamma) moderately enhanced TNF-alpha production. Our results indicate that monocytes are mainly responsible for the production of TNF-alpha in response to allogeneic stimulation, and T-cells modulate monocyte function by their soluble factors. PMID- 8429685 TI - Isolation and characterisation of a CDw50 negative Jurkat T-cell line variant (PPL.1). AB - PPL.1, a Jurkat cell line variant deficient in CDw50 surface expression, has been selected by fluorescence-activated cell sorting and expanded in cell culture. We have studied the expression of several leukocyte surface markers (CD2, CD3, CD4, CD8, CD7, CD26, CD25, CD14, CD18, kCD20, CD43, CD45, CD45R, CD71 and HLA class I and II) and we find no differences in their expression between PPL.1 and its parental Jurkat cell line. Immunoprecipitation analysis of metabolically labelled PPL.1 cells ([35S]-cysteine plus [35S]-methionine) fails to detect the presence of a preformed cytoplasmic pool of CDw50 molecules. The deficient CDw50 expression on PPL.1 cells is stable after several weeks of continuous culture and even after exposure of cells to several lymphocyte activating agents (PGE2, PHA, Con A, calcium ionophore A23187 and human recombinant IFN-gamma). No karyotype changes responsible for such phenotype deficiency are found. PPL.1 cells are as efficient as wild-type Jurkat or K562 cells, when used as targets in cytotoxicity assays with fresh or PHA-stimulated peripheral blood lymphocytes. No blocking effects of CDw50-specific mAb are observed in such assay. These results are consistent with the fact that CDw50 is not involved in alloreactive T-cell specific cytotoxicity. They also suggest that this antigen is implicated only on a very specialized type of cell-cell interactions. PMID- 8429686 TI - The expression of tissue factor antigen and activity on the surface of leukemic cells. AB - Tissue factor activity of intact cell and cell lysate, and the presence of tissue factor antigen on cell surface, were examined in leukemic cells from patients with acute myelogenous leukemia (AML, M1-M5) or acute lymphoblastic leukemia (ALL L1), and in mononuclear cells from normal donors. Leukemic cells from AML or ALL had significantly more tissue factor activity not only on intact cells but also in cell lysate than mononuclear cells from normal donors (p < 0.001). Tissue factor activities of the intact leukemic cells and lysate from AML patients with DIC were significantly higher than those without DIC (p < 0.001). The relationship between the percent of positive cells for tissue factor and the presence of DIC at the time of diagnosis of acute leukemia was observed. The patients with DIC showed the higher percentage of tissue factor-positive cells than those without (p < 0.01). The development of DIC following chemotherapy was recognized in 2 out of 7 AML-MI patients and 2 out of 4 ALL-L1 patients who had relatively high tissue factor activities of cell lysate. The release of tissue factor from cytoplasm induced by chemotherapy would be another mechanism for the development of DIC. The report suggests the possibility of the prediction for DIC by the flowcytometric assay of tissue factor antigen. PMID- 8429687 TI - Effects of carboplatin in combination with other anticancer agents on human leukemia cell lines. AB - In vitro studies with drug combinations of carboplatin and other anticancer agents were carried out in MOLT-3 human lymphoblastic leukemia and HL-60 human promyelocytic leukemia cell lines. Cells were incubated for 3 days in the presence of various concentrations of carboplatin and other drugs and cell growth inhibition was determined by MTT assay. The antitumor effects of the drug combinations at ID50 were analyzed using an improved isobologram. In MOLT-3 cells, supra-additive (synergistic) effects were observed for carboplatin in combination with cytosine arabinoside, mitoxantrone and CPT-11. Additive effects were observed for combinations of carboplatin with bleomycin, daunorubicin, doxorubicin, etoposide, 6-mercaptopurine, and vincristine. Sub-additive and protective (antagonistic) effects were observed with methotrexate. Synergistic or antagonistic effects for combinations of carboplatin and CPT-11, cytosine arabinoside, mitoxantrone and methotrexate were also observed in HL-60 cells. These findings suggest that carboplatin has additive or synergistic cytotoxic effects with most of the agents tested. Determination of the usefulness of these drug combinations awaits appropriate in vivo experiments that should assess both tumorcidal effects and possible increased toxicity. The simultaneous administration of carboplatin and methotrexate would be of little effect. To find optimal schedules for this combination, further pre-clinical studies of various combinations schedule would appear to be warranted. PMID- 8429688 TI - The use of an enzyme single fiber reactor in the study of leukemic cell proliferation: in vitro experiments and computer simulation. AB - This paper describes the use of an immobilized enzyme reactor in the study of the in vitro effects of lysine deprivation on leukemic blood. L-lysine alpha-oxidase is immobilized in a single hollow fiber reactor to remove lysine from the blood of sheep infected by BLV. The treatment relies on the higher sensitivity of leukemic cells to nutrient depletion than that of normal cells. A population balance model is used to describe the changes in the leukocyte proliferative capacity after treatment. Additionally, preliminary data from in vitro tests with human blood demonstrate the potential of L-lysine alpha-oxidase and the enzymatic reactor in treating leukemia. PMID- 8429689 TI - Up-regulation of small GTP-binding proteins smg P21A and ras P21S during TPA induced differentiation of human leukemia cell lines. AB - The protein smg p21A/Krev-1/rap 1A was identified as a ras p21-like small G protein, having the ability to revert v-Ki-ras transformed NIH 3T3 fibroblasts. The expression level of smg p21A and ras p21s during phorbol ester-induced differentiation of HL-60 and MEG-01 cell lines was analyzed by immuno- and Northern blotting. In both cell lines, levels of smg p21 and ras p21s increased quickly in early phase of differentiation along with the appearance of differentiation phenotypes. They increased 3-4 fold on days 1-2, then decreased gradually. The increasing smg 21 mRNA levels also corresponded with that of products. Among ras mRNAs, Ha-ras and N-ras transcripts increased somewhat faster than smg 21. These small G-proteins may play closely related roles in the differentiation of these leukemia cell lines. PMID- 8429690 TI - Effects of interleukin 2 on myelodysplastic syndromes. AB - To evaluate the clinical usefulness of interleukin 2 (IL-2) on myelodysplastic syndromes (MDS), the serum IL-2 level, the effect of IL-2 on the proliferation of blasts, and the cell-mediated cytotoxic effect of IL-2 on blasts were examined in MDS patients. Of 18 patients, 2 patients had an increased serum IL-2 level. Although the proliferation of blasts in most cases, including the two patients having a high serum IL-2 level, was not stimulated by IL-2, the blasts of one case apparently proliferated in response to IL-2. It was also clearly shown that IL-2-stimulated normal peripheral blood mononuclear cells (PBMNC) showed cytotoxicity against MDS blasts, whereas the PBMNC of the advanced stages of MDS were usually defective in regard to this IL-2-dependent cytotoxicity. The therapeutic usefulness of IL-2 or lymphokine-activated killer cells for MDS was not established by the present study. PMID- 8429691 TI - Adsorption and uptake of the alkyllysophospholipid ET-18-OCH3 by HL-60 cells during induction of differentiation by dimethylsulfoxide. AB - The human leukemic cell line HL-60 is highly sensitive to the antineoplastic agent alkyllysophospholipid, 1-octadecyl-2-methyl-sn-glycerol-3-phosphocholine (ET-18-OCH3). We investigated the adsorption and uptake of radiolabeled ET-18 OCH3 in undifferentiated HL-60 cells and during differentiation to granulocytes induced by dimethylsulfoxide. HL-60 cells become less sensitive to the cytotoxic action of ET-18-OCH3 during differentiation. The decrease in sensitivity is correlated with a decrease in both adsorption and uptake of [3H]ET-18-OCH3 during differentiation. Binding studies revealed that the binding of ET-18-OCH3 to both undifferentiated and differentiated HL-60 cells is non-saturable which renders the existence of a specific binding place highly unlikely. PMID- 8429692 TI - Characteristics of chemotherapy-induced clinical remission in long survivors with aggressive adult T-cell leukemia/lymphoma. AB - The acute and lymphoma types of adult T-cell leukemia/lymphoma (ATL) usually have a very poor prognosis, although some patients achieve long survival after chemotherapy. A total of 114 patients with these aggressive types of ATL were newly diagnosed at our institution from 1975 to 1989. By multivariate analysis, poor performance status and high serum creatine levels were associated with shortened survival. With combination chemotherapy, 20 patients achieved complete remission (CR), 53 achieved partial remission (PR) and 35 showed no response. Fifteen of the CR or PR patients survived for more than two years and all other patients survived for less than two years. As compared with short survivors (< 2 years) after remission, long survivors (> or = 2 years) after remission had a higher CR/PR ratio, a longer time until remission and a higher doxorubicin dose to achieve remission. Death due to causes other than the primary disease occurred in 18% of short survivors after remission and in 11.2% of nonresponders, but in none of the long survivors. Long survivors with acute ATL included 6 patients with CR and 5 patients with PR. All four lymphoma type ATL long survivors achieved CR. Monoclonal integration of HTLV-I provirus was detected in the peripheral blood mononuclear cells of all 3 PR long survivors with acute ATL studied, but was not detected in all 4 CR cases studied at remission. The minimum CD4/CD8 ratio of peripheral mononuclear cells at remission was < 1.0 in all acute ATL long survivors with CR, and was > 1.0 in all acute ATL long survivors with PR. Three out of six acute ATL long survivors with CR developed suspected viral infection just before achieving CR. Our findings show that in aggressive ATL the characteristics of remission are heterogeneous even among long survivors. PMID- 8429693 TI - Deoxyribonucleoside triphosphate pools and thymidine chemosensitization in human T-cell leukemia. AB - Thymidine kills cells by depleting dCTP stores. The present experiments tested whether deoxycytidine, by replenishing dCTP pools, could prevent thymidine cytotoxicity and thymidine's enhancement of carboplatin killing in two human T cell acute leukemia cell lines. MOLT3 and JM cells were exposed to combinations of thymidine, deoxycytidine, and carboplatin and then assessed for survival, the magnitude of thymidine-carboplatin chemosensitization, and changes in deoxyribonucleoside triphosphate pools. For both cell lines, deoxycytidine (up to 144.5 micrograms/ml x 24 h) completely restored dCTP pools but only partially protected against thymidine cytotoxicity (100-1000 micrograms/ml x 24 h) and thymidine-carboplatin sensitization (up to 60 micrograms carboplatin/ml during the last hour of thymidine). This contrasts with complete protection in prior studies using other cell types. Thymidine alone markedly increased dTTP and dGTP pools and decreased dCTP; dATP pools underwent a sharp decline which has not been observed before in any cell line. In subsequent studies 0.0336-137.3 micrograms deoxyadenosine/ml partially prevented cytotoxicity and carboplatin sensitization by 300 micrograms thymidine/ml. Together, deoxycytidine and deoxyadenosine completely prevented thymidine-carboplatin sensitization even though dATP and dCTP pools were not entirely returned to normal. These findings are discussed in regard to the unusual sensitivity of T-cell malignancies to thymidine toxicity, mechanisms of cytotoxicity and chemosensitization by thymidine, and the possibility of thymidine selectively sensitizing T-cell malignancies to killing by alkylating agents. PMID- 8429694 TI - Intervention treatment of established neutropenia with human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF) in patients undergoing cancer chemotherapy. AB - Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) was given to 60 patients, in a double-blind, non-prophylactic study of already established chemotherapy-induced leucopenia, for 5 days by continuous intravenous infusion and twice or once daily by subcutaneous injection. Four patients were randomized to rhGM-CSF (3) or placebo (1) at each dose (1.3, 1.7, 5.5, 11, or 22 micrograms of protein/kg). Leucocyte recovery was significantly enhanced compared with controls, in a dose-dependent manner except for 22 micrograms/kg which was ineffective with a worse experience of side effects in some patients. Most adverse events occurred in equal proportions in the treated and placebo cases. Fourteen patients developed infection and were treated with antibiotics in addition to rhGM-CSF. They were joined by a further 18 febrile patients and treated with rhGM-CSF in a subsequent open-label trial. The survival from infection was related to white blood cell (WBC) count: 19 of 32 responded with increased numbers of leucocytes (WBC count above 1.5 x 10(9)/1) after 5 days of GM-CSF. Sixteen of the 19 leucocyte 'responders' recovered from infection, two died from the underlying disease and one from persistent infection. Six of the 13 patients who did not have a leucocyte response died with persistent infection. These data indicate that rhGM-CSF enhances the leucocyte count following chemotherapy and in this way saves critically ill neutropenic patients from fatal infections. PMID- 8429695 TI - Leukemia in newborn infants with Down syndrome. PMID- 8429696 TI - [Comparative study of protected alveolar lavage versus occluded telescopic catheter in patients with suspected pneumonia and under mechanic ventilation]. AB - BACKGROUND: The pneumonias associated to mechanical ventilation present great difficulty in diagnosis and have a high mortality. The invasive diagnostic technique of choice in these patients is bronchial curettage by a double telescopic catheter with distal occlusion (OTC) based on its good sensitivity/specificity relation. Recently, the use of a variant of the classical bronchoalveolar lavage (BRL), bronchoalveolar lavage or protected alveolar lavage (PAL) has appeared in the diagnosis of conventional bacterial pneumonia. This new technique provides good specificity of OTC by its use with "protected" catheters and a high sensitivity due to exploration of a greater area of the lung. METHODS: Twenty patients receiving mechanical ventilation (MV) suspected of pneumonia in whom 21 fibrobronchoscopies (FB) were performed with OTC and PAL were studied with quantification of the cultures obtained being carried out. The OTC was performed according to the usual technique and PAL by the instillation of 40 ml of saline serum administered through a Combicath type catheter. RESULTS: OTC and PAL provided diagnostic results which coincided in 8 cases: the same germs were isolated at significant concentrations in six patients and in the two remaining cases direct immunofluorescence for Legionella was positive. PAL was diagnosed in 4 more cases with the diagnosis of viral inclusion bodies being possible in one upon cytologic examination. The count of cells with intracellular bacteria (ICB) was greater than 7% and was always related with positivity in the PAL. CONCLUSIONS: A greater sensitivity was observed with the protected alveolar lavage technique. Moreover, this technique makes virologic investigation and the counting of cells with intracellular bacteria, which may be a marker of rapid diagnosis of bacterial pneumonia, possible. PMID- 8429697 TI - [Heart arrest in outpatients resuscitated without neurologic sequelae]. AB - BACKGROUND: Around 65% of the deaths by acute myocardial infarction are presented in the first hour of the attack when most patients are outside a hospital and it is almost always caused by ventricular fibrillation. Home defibrillation permits these patients to be saved if an integral emergency system is available. METHODS: The cases of ambulatory cardiac arrest resuscitated with no residual neurologic lesions in the province of Ciudad Real were reviewed. The hospital possesses an integral emergency department with an area of communication with one sole emergency telephone (006) and 7 mobile intensive care medical units. RESULTS: Five male patients with a mean age of 54 years were ambulatory defibrillated (4 at home and 1 at work) following cardiac arrest by ventricular fibrillation of acute myocardial infarction (4 inferior localization and 1 anterior). All the patients called for angina, except one for cardiorespiratory arrest. The mean time of delay between the call for help and the arrival of the doctor was 8.4 +/- 6.3 minutes; ventricular fibrillation was witnessed by a doctor in all the cases except for one. Ventricular fibrillation was recurrent in three patients and all required more than one defibrillation. Two patients required mechanical ventilation and vasoactive drugs due to cardiac failure. The complications secondary to resuscitation manoevers are commented upon but the absence of neurologic sequelae is of note. All the patients were discharged and returned to work with the exception of one patient who died due to rupture of the posteromedian muscle of the mitral valve. CONCLUSIONS: Cardiac arrest by ventricular fibrillation in the acute myocardial infarction may be successfully treated out of the hospital with an integral emergency system. PMID- 8429698 TI - [Medicina Clinica: 50 years]. PMID- 8429699 TI - [Bronchoalveolar lavage in the diagnosis of pulmonary infection]. PMID- 8429700 TI - [Alcoholism: a black hole in our medical training]. PMID- 8429701 TI - [Differential aspects of drug prescriptions in elderly people living in the northeast area of Granada: study of 366 individuals]. AB - BACKGROUND: The geriatric population consumes greater numbers of medicines than other age groups. We compared drug prescription and consumption in elderly persons residing in a nursing home or living in their own home. METHODS: We studied 366 elderly men and women (older than 65 years) who lived either in nursing homes or in their own home. The variables recorded were the most frequent diseases, number and types of drugs prescribed per person, and adverse reactions of the most frequently prescribed drugs. RESULTS: The most common diseases recorded among subjects living at home were arterial hypertension and rheumatic diseases; among those in nursing homes, the most common ailment was dementia. Subjects in nursing homes were prescribed more different drugs (2.8 +/- 1/person) than individuals living on their own (1.8 +/- 1/person). The percentage of adverse reactions was highest in ambulatory patients (64%) and in patients taking benzodiazepines (56%). A large proportion of all prescriptions (74.5%) were for solid galenic forms. CONCLUSIONS: We draw attention to the greater consumption of drugs by elderly persons residing in nursing homes, as well as the prescription of benzodiazepines and analgesic-antiinflammatory drugs within old age. PMID- 8429702 TI - [Genetic markers in neuropsychiatric disorders: genetic cartography and diagnostic and therapeutic explorations (II)]. PMID- 8429703 TI - [High output heart insufficiency as presentation form of multiple myeloma]. PMID- 8429704 TI - [Zollinger-Ellison syndrome with basal gastrin and non-diagnostic stimulation tests (secretin and calcium)]. PMID- 8429705 TI - [Antiphospholipid antibodies associated with epilepsy, atypical migraine, and chorea]. PMID- 8429706 TI - [Sudden death, exercise, and blood potassium]. PMID- 8429707 TI - [Relationship between nosocomial infection and hospital mortality. Multicenter study]. AB - BACKGROUND: Hospital mortality related to infections acquired in the hospital setting has not been well studied in Spain. We carried out a study of seven hospitals in order to assess and quantify the problem. METHODS: The study period included three months of observations (between November 1, 1989 and January 31, 1990), and data pertaining to all deaths of patients hospitalized for a minimum of 24 hours were collected. The number of people admitted within the study period was 16,025, and the number of deaths registered and included in our study was 488 (3%). The data were obtained from the patient's medical history one week after death as well as from the hospital physicians on the case. In order to quantify the interobserver variability derived from the classification criteria, the simple kappa index was calculated and averaged to form an ordinal scale. RESULTS: 216 (44.3%) of 488 deaths included in our study had no infection, 138 (28%) had an infection no-hospital-associated, and 134 (27%) had nosocomial infection (50 10%--"causally related to death", 59-12%--"contributing to death", and 25-5%- "not related to death"). The lower respiratory tract infections, bacteremias and surgical wound infections were the most related to cause of death. Staphylococcus aureus was the pathogen most frequently associated with the infections found at the time of death. CONCLUSIONS: Those patients admitted with non-fatal diseases made up the greatest percentage (39.9%) of deaths from nosocomial infections. The infection was considered the direct cause of death in 18.8% of these cases, although the differences found had no statistical significance. PMID- 8429708 TI - [Pharmacokinetic study of zidovudine in parenteral drug addicts with human immunodeficiency virus infection]. AB - BACKGROUND: The aim of this study was to know the pharmacokinetics of zidovudine (ZDV) in steady state in patients with infection by the human immunodeficiency virus (HIV) in whom the risk factor was intravenous drug use. METHODS: The study was carried out in 8 patients in stage IV of the Centers for Disease Control (CDC) with no acute intercurrent process, with normal liver and renal function, orally receiving 250 mg of ZDV every 6 hours. Blood samples were taken between 30 and 360 minutes from the last doses taken during fasting. Plasma concentrations of ZDV and glucuronide zidovudine (G-ZDV) were determined by radioimmunoassay with the data being adapted to a monocompartmental pharmacokinetic model. RESULTS: The maximum concentration (Cmax) of ZDV was 0.81 +/- 0.38 microgram/ml demonstrating high interindividual variability with values between 0.35 microgram/ml and 1.45 microgram/ml. The mean Cmax of G-ZDV was 1.44 +/- 0.64 microgram/ml. The mean t1/2 of ZDV and G-ZDV was 1.63 +/- 0.75 hours and 1.12 +/- 0.32 hours, respectively, with values oscillating between 0.99 and 3.14 h in the case of ZDV. The area below the curve concentration/time (AUC) of ZDV was 1.43 +/ 0.34 microgram-h/ml and in the case of G-ZDV the AUC was 2.73 +/- 0.91 microgram h/ml. Total body clearance (Clb) of ZDV was 2.11 +/- 0.64 l/kg/h and the volume of distribution (Vd) of ZDV was 5.6 +/- 1.73 l/kg. CONCLUSIONS: The results of this study demonstrate that there is a marked interindividual variability in the pharmacokinetics of zidovudine suggesting the need for adapting dosage to patients weight and monitoring plasma concentration. PMID- 8429709 TI - [Prevalence of antibodies against hepatitis C virus in a sample of homosexual males]. AB - BACKGROUND: The finding of an important proportion of cases of hepatitis C without previous contact with blood or hemoderivates has led to suspect that there may be other routes of transmission among which sexual transmission may be found. METHODS: The presence of antibodies against the hepatitis C virus (anti HCV) and the association of this infection with certain epidemiologic parameters and sexual practice was determined in 184 homosexual males with no other risk factors for virasis of intravenous transmission. Moreover, the prevalence of anti HCV was evaluated in 210 voluntary blood donors. Every homosexual was surveyed on sexual practice and the first serum sample available of each of these patients was analyzed for anti-HVC and anti-HIV-1. RESULTS: Twenty-two homosexual (12%) were anti-HCV positive while only one (0.5%) of the control was positive for this marker (p < 0.0001). Thirty-six cases (19%) were anti-HIV positive (none of the controls). No statistical association was found between both serologic markers or between the presence of anti-HCV and the epidemiologic parameters evaluated. CONCLUSIONS: Homosexuals constitute a risk group for hepatitis C virus infection although the sexual route is probably not effective for transmission of this entity. PMID- 8429710 TI - [Surveillance of nosocomial infection at a regional++ hospital. Results of incidence and prevalence studies in a 2 years' experience]. AB - BACKGROUND: The most appropriate methods for surveillance of nosocomial infection (NI) in hospitals with less than 250 beds remain to be elucidated. The aims of this study were to investigate the differences between the results found in a study in incidence (IS) and in another concerning prevalence (PS) carried out in a county hospital and study the experience in the application of a collection method of incidental cases through a survey. METHODS: For 2 years IS trimestrally accumulated and simultaneously PS trimestrally were applied for global surveillance and for type of NI as well as for knowing the etiology of the same. In both types of study all the hospital patients were included. RESULTS: The accumulated incidence of patients with NI over the 2 years was 3.9% (3.6 in the first 12 months and 4.1 in the 12 remaining months) and the global prevalence was 7.5%. The global trend of NI was that of an increase in both studies. The most frequent NI in both studies were surgical wound and urinary infection. PS did not detect the least frequent NI. The etiologic agents of NI were similar in both studies. The survey undertaken for the detection of NI demonstrated 61% sensitivity, 98% specificity and a positive prediction value of 94%. CONCLUSIONS: In small hospitals global prevalence studies may be useful for surveillance of the most frequent nosocomial infections. The results obtained by incidence studies were less variable and better reflect the trend of nosocomial infection. A survey used as a study method of incidence for the collection of cases of nosocomial infection loses sensitivity over time. PMID- 8429711 TI - [Dangers of silicone breast prosthesis]. PMID- 8429712 TI - [Bartter's syndrome: variability and clinical course]. AB - The clinical, hydroelectrolytical and hormonal features, as well as the treatment of 3 patients with Bartter's syndrome are analyzed. The patients consulted due to severe hypokalaemia with little clinical expression (paresthesiae, carpopedal spasms, polyuria, polydipsia and/or weakness). All had normal blood pressure, hypokalemia, hyperreninaemic hyperaldosteronism and a decrease in the fractional distal chloride reabsorption; hyperuricaemia was observed in two cases and hypomagnesemia in one. Treatment with oral potassium supplements and indomethacin managed to raise serum K to around the lower limit of the normal range. However, the tubular defect and hyperreninaemic hyperaldosteronism persisted. The urinary excretion of prostaglandin E2 was normal in both the patients in whom it was measured. Given the minimal clinical expression of hypokalemia despite the difficulty in correcting it, it is probable that this syndrome often remains unrecognized. PMID- 8429713 TI - [Bone marrow transplantation: indications, methods, and results]. PMID- 8429714 TI - [Value of the Karnofsky index as indicator of life prognosis in terminal oncology patients]. PMID- 8429715 TI - [Unusual factor of decompensation of epilepsy]. PMID- 8429716 TI - [Hyperreactive malarial splenomegaly]. PMID- 8429717 TI - [Neutrophilic leukemoid reaction and pleural mesothelioma]. PMID- 8429718 TI - [The day that I did not kill Mikado]. PMID- 8429719 TI - [The year 2000: health for all and power for a few?]. PMID- 8429720 TI - [Weight loss in the treatment of obstructive sleep apnea syndrome]. PMID- 8429721 TI - [Ethics and scientific politics]. PMID- 8429722 TI - [Skin lymphocytoma caused by Borrelia?]. PMID- 8429723 TI - [Pregnancy and thrombophilia in women with congenital deficit of antithrombin III, protein C, protein S or plasminogen: analysis of 39 cases]. AB - BACKGROUND: Pregnancy, delivery and puerperium are situations which increment the risk of thromboembolic complications in women who are carriers of congenital heterozygotic deficits of type I antithrombin III (ATIII), protein C (PC) or protein S (PS). The aim of this study was to analyze the experience of the authors and propose therapeutic conduct in each case. Furthermore, the spontaneous losses of pregnancy related with these deficits were studied. METHODS: Thirty-nine women, seventeen with ATIII deficit, fifteen with PC deficit and four with a deficit of PS and three with a plasminogen (Pg) deficit totalling 79 pregnancies and 51 thrombotic episodes sixteen of which were related with the pregnancy, delivery or puerperium were studied. The antigenic and functional activity of ATIII, PC, PS and Pg were determined. RESULTS: The incidence of thrombosis for the ATIII deficit during pregnancy was 39%, which was greater, of statistical significance (p = 0.046), than the 15% observed during puerperium. In women with a deficit of PC, the incidence of thrombosis was 4.5% during pregnancy and 14% during puerperium with no significant difference between the two situations. The incidence of thrombosis during pregnancy and postpartum in the deficit of ATIII was significantly higher (p < 0.025) than that observed for the deficit of PC. For women with a deficit of PS and Pg the incidence of thrombosis was nul in pregnancy and puerperium. CONCLUSIONS: Pregnancy and puerperium are situations which trigger thrombotic phenomena and increase the risk of the same in women with a deficit of antithrombin III and protein C and, to a lesser degree, the deficit of protein S or plasminogen. A strict control of these situations and individualized treatment is required according to the type of deficit, presence of previous thromboembolic history and anticoagulant history at the time of pregnancy. No increase in the risk of loss of pregnancy in any of the deficits studied was observed. PMID- 8429724 TI - [Clinical epidemiology of an outbreak of nosocomial infection caused by Staphylococcus aureus resistant to methicillin and aminoglycosides: efficacy of control measures. Comite de Control de Infecciones]. AB - BACKGROUND: The appearance of outbreaks of nosocomial infections due to methicillin resistant Staphylococcus aureus (MRSA) is a current problem in Spain. The clinical and molecular epidemiology of these outbreaks as well as the efficacy of their control measures are a matter of controversy. METHODS: An outbreak of MRSA nosocomial infections in the Hospital Clinic i Provincial of Barcelona, a 953-bed University Hospital, with a total of 347 cases from September 1989 to October 1991 is described. The control measures used include prospective and continued surveillance of all MRSA isolations, identification of the reservoir, use of different types of isolation and control of nasal carriers among health care workers. The MRSA strains isolated were studied by standard microbiological procedures, phage-typing and extrachromosomal (plasmid) DNA analysis by means of restriction endonuclease analysis (REAP). RESULTS: From the Intensive Care Units, the outbreak extended to the medical and surgical wards. Seventy-one percent of the cases corresponded to infected patients and 29% to asymptomatic carrier patients. The MRSA strain responsible of the outbreak had a notable antibiotic multiple-resistance pattern. The studies performed showed that most of the strains belonged to phage group III and were lysed by phage 77. Plasmid DNA analysis showed that 95% of the strains isolated had a unique homogeneous profile. Despite the different control measures used, the MRSA infection has acquired a medium level endemic rate in the Hospital Clinic i Provincial. CONCLUSIONS: The introduction and spread of methicillin-resistant MRSA in a teaching hospital with more than 500-bed may be rapid and affecting large number of patients. Effective control measures carries multiple problems, which must be addressed with the collaboration of all hospital employees. The molecular typing techniques used (REAP) further identified that the outbreak is due to one single MRSA strain, with an epidemiologic behavior identical to the one showed by the epidemic strains previously described in the United Kingdom and Australia. PMID- 8429725 TI - [Postoperative psychiatric complications following liver transplantation]. AB - BACKGROUND: The aim of the study was to describe the psychiatric complications of the postoperative period of liver transplantation. METHODS: Sixty-three adult liver transplant recipients were seen weekly before surgery and during the postoperative period until discharge. They were assessed through Goldberg's Clinical Interview Schedule (CIS) and diagnosed according to DSM-III-R criteria. RESULTS: Psychiatric morbidity was 29%. Organic mental disorders were the most prevalent, especially delirium (13%), followed by adjustment disorders (8%), major depression (5%) and organic anxiety disorders (3%). One of the depressed patients showed subsequently a rapidly cycling bipolar course. Most patients had a good outcome of their psychiatric disorder at discharge. In contrast, patients with delirium presented a higher mortality rate. CONCLUSIONS: The prevalence of short-term psychiatric complications in patients undergoing liver transplantation is close to 30%. Delirium, which is significantly related to mortality, is the most frequent of them, followed by adjustment disorders and affective disorders, which, in contrast, seem to have a good prognosis. PMID- 8429726 TI - [Emphysema caused by alpha 1-antitrypsin deficit. New preventive and therapeutic measures]. PMID- 8429727 TI - [Primary thrombophilia and pregnancy]. PMID- 8429728 TI - [Lymphomatoid papulosis evolving into malignant lymphoma. Clinico-pathological and immunohistochemical study of 3 cases]. AB - Lymphomatoid papulosis is a disorder characterized by recurrent skin lesions with histological features suggestive of malignant lymphoma. In most cases the cutaneous lesions heal spontaneously but the course of the disease is long lasting and an evolution into a Hodgkin's disease and non-Hodgkin lymphomas may be seen. We report herein the clinical, pathological and immunohistochemical study of three patients having a long-standing lymphomatoid papulosis, which turned into Hodgkin's disease, mycosis fungoides and nodular paragranuloma, respectively. Immunohistochemical analysis showed that the immunophenotype of atypical cells in lymphomatoid papulosis was similar to that observed in Reed Sternberg cells in Hodgkin's disease and the neoplastic cells of mycosis fungoides. However, the immunohistochemical profile of cells in lymphomatoid papulosis differed from those observed in cells of nodular paragranuloma, developed by one of the three patients. The relationship between lymphomatoid papulosis and malignant lymphomas associated to lymphomatoid papulosis is discussed. The results show that no definitive criteria can be infered from an immunohistochemical study in lymphomatoid papulosis, in predicting the clinical evolution of the disease. PMID- 8429729 TI - [Idiopathic juvenile osteoporosis]. PMID- 8429730 TI - [Sclerosing carcinoma of the liver. Report of a case diagnosed in vivo]. PMID- 8429731 TI - [A hospital information service]. PMID- 8429732 TI - [Analysis of Spanish biomedical periodicals and their impact]. PMID- 8429733 TI - [Candida albicans esophagitis in AIDS patients]. PMID- 8429734 TI - [Filgrastim (rG-CSF) and neutropenia associated with AIDS, is MEDLINE lying?]. PMID- 8429735 TI - [Elevation of HIV-antigen concentration during administration of zidovudine]. PMID- 8429736 TI - [Why are there regional differences in drug utilization? Explanation from a clinical pharmacologist]. PMID- 8429737 TI - [University hospital--an academy. Concentrating research resources]. PMID- 8429738 TI - [A suitable prioritization?]. PMID- 8429739 TI - [Good pacemaker care is possible even at small hospitals]. PMID- 8429740 TI - [No reasons to change iodine-131 treatment]. PMID- 8429741 TI - [No nicotine patch to pregnant women!]. PMID- 8429742 TI - [No strict age limits]. PMID- 8429743 TI - [Reduced number of cases of hereditary mental retardation]. PMID- 8429744 TI - [Syphilis in the USA in the shade of AIDS. Most frequent among coloured crack addicts]. PMID- 8429745 TI - [Autoimmunity. A highly specialized immunologic reaction or a manifestation of chaos?]. PMID- 8429746 TI - [The T-lymphocyte. A hero or a villain in the drama?]. PMID- 8429747 TI - [Determine c-ANCA in subglottic stenosis]. PMID- 8429748 TI - [Marfan syndrome and pregnancy]. PMID- 8429749 TI - [Mitomycin C caused contact allergy]. PMID- 8429750 TI - [Bouveret syndrome. Unusual cause of gallstone ileus]. PMID- 8429751 TI - [The man behind the syndrome: Leon Bouveret. The internist who supported surgery]. PMID- 8429752 TI - [Three anesthesia departments scrutinize each other. Accreditation according to an Australian model]. PMID- 8429753 TI - [Theses on patient care ideology in the light of ethics. Comparison between primary health care ideology and hospital ideology]. PMID- 8429754 TI - [First results from a randomized study: ABC for alternative childbirth]. PMID- 8429755 TI - H1 contractile and H2 relaxant receptors in canine gastric muscularis mucosae. AB - Histamine had two effects on the contractility of canine gastric muscularis mucosae in vitro: relaxation at or below 10 microM and contraction at higher concentrations. Selective agonists and antagonists were used to test the possibility that these effects were mediated by different receptor subtypes. The H1-selective agonist 2-pyridylethylamine (2-PEA) and the H2-selective agonist dimaprit contracted and relaxed this muscle, respectively, while the H3-selective agonist R-alpha-methylhistamine had no effect. The H1- and H2-selective antagonists mepyramine and ranitidine selectively blocked 2-PEA-mediated contractions and dimaprit-mediated relaxations, respectively. Agonist responses, were unaltered by tetrodotoxin, suggesting a site of action other than nerves. Our results indicate that canine gastric corpus muscularis mucosae possesses both contractile H1 and relaxant H2 receptors. PMID- 8429756 TI - Muscarinic receptors after syngeneic unilateral lung transplantation. AB - We have characterized the muscarinic cholinergic receptor population by binding assay in left and right lung membranes from syngeneic Lewis rats one month after unilateral left lung transplantation and from controls. The density of muscarinic receptors measured by [3H](-)QNB binding was similar in left and right lung membranes one month after unilateral left lung transplantation (Bmax = 28.2 +/- 2.2 and 29.0 +/- 1.4 fmol/mg protein, respectively) and was similar between transplanted and control rats. Binding affinity also was similar on both sides (Kd = 28.5 +/- 9.5 and 32.3 +/- 2.7 pM, respectively). Binding parameters (Ki) of four selective muscarinic antagonists determined from competitive binding experiments of 0.5 nM [3H](-)QNB showed an order of potency: atropine > 4-DAMP > AF-DX 116 BS > pirenzepine both in transplanted and in control lungs. There was no significant difference in Ki values between left transplanted and right non transplanted lungs (1.8 +/- 0.1 and 1.9 +/- 0.1 nM; 4.3 +/- 0.5 and 3.5 +/- 0.5 nM; 80.9 +/- 13.8 and 78.9 +/- 18.8 nM; 480.4 +/- 40.9 and 481.7 +/- 78.2 nM, respectively for atropine, 4-DAMP, AF-DX 116 BS and pirenzepine). Values were also similar between transplanted and control lungs. These observations suggest a similar population of muscarinic receptors in the transplanted and the non transplanted lung in the Lewis rat. This result is in accordance with the similar physiological effect of exogenous acetylcholine that we previously observed in transplanted and non transplanted bronchi. PMID- 8429757 TI - Lipolytic effect of beta-endorphin in human fat cells. AB - Recently, a role of beta-Endorphin on peripheral tissue metabolism has been suggested. A lipolytic effect of beta-Endorphin has been observed both in vivo and in vitro in animals but, at present, there is no evidence for a similar effect in humans. In this study, we investigated the lipolytic effect of beta Endorphin in isolated human adipocytes. beta-Endorphin induced a significant increase in glycerol release in isolated human fat cells. Naloxone was able to inhibit the beta-Endorphin-induced lipolysis. The opioid antagonist alone had no effect on basal lipolysis and on Epinephrine-stimulated lipolysis when administered together with this hormone. Our results suggest that beta-Endorphin may play a role on lipolysis also in human fat cells and that this effect may be mediated by a specific opiate receptor. PMID- 8429758 TI - Absence of monoclonal antibody detectable Kaposi sarcoma-specific antigens on lesion-derived cultured cells. AB - To define the histogenesis and cell origin of Kaposi sarcoma (KS), we cultured KS cells without retrovirally conditioned media from three HIV seropositive AIDS patients and then attempted to raise mouse hybrid monoclonal antibodies (Mabs) specific to these AIDS-KS cells. After both in vivo and in vitro immunization trials, all putative Mabs reacted positively to KS cells but also non specifically with other human (CH5 and OM) and non-human (RSE-1) control endothelial cell lines. To overcome this crossreactivity, we further "absorbed" previously cloned hybrids and pre-hybrid splenocytes by incubating them with the control endothelial cell lines to eliminate splenocytes and/or hybridomas reactive to normal endothelium. Whereas absorption successfully eliminated immunoreactivity to control endothelium, it also excluded reactivity to KS cells. These findings (lack of specific antigenicity and immunoresponsiveness of KS similar to non-KS control endothelium) suggest that AIDS-KS cells are neither antigenically transformed nor neoplastic, but instead represent dedifferentiated or transdifferentiated endothelium which retains immunogenicity of its original endothelial cell prototype. PMID- 8429759 TI - Evidence for 5-HT2 receptor involvement in the stimulation of preovulatory LH and prolactin release and ovulation in normal cycling rats. AB - The participation of serotonin (5-HT) in the control of LH secretion and ovulation has been reported in numerous pharmacological experiments, although the results have been contradictory. In this study to determine the physiological involvement of 5-HT2 receptors in the control of preovulatory LH and prolactin (PRL) release and ovulation, ketanserin (KET), a selective 5-HT2 receptor antagonist, was administered to normal cycling rats. Oral administration of KET at 10 mg/kg BW between 0900h and 1300h on the day of proestrus completely blocked ovulation, while the administration after 1500h did not. The ovulation was confirmed 2 days after treatment with KET. There was no consistent effect on the number of ova ovulated. Co-administration of KET with 5-methoxy-N,N dimethyltryptamine (5-MeDMT), a 5-HT agonist, at a dose of 2 mg/kg BW ip reversed KET-induced inhibition of ovulation. The highest rate (89%) of reversal was obtained by co-administration at 1500h. KET administered at 1100h completely blocked the preovulatory surge of LH and PRL observed in control animals at 1800h. The present study suggests that 5-HT stimulates preovulatory LH and PRL surge and ovulation via 5-HT2 receptor under physiological conditions and that the effect of 5-HT depends on the critical period and the 24-hour periodicity. PMID- 8429760 TI - Estradiol increases glucocorticoid binding and glucocorticoid induction of ornithine decarboxylase in the rat spinal cord. AB - Previous results demonstrated that estradiol (E2) treatment of ovariectomized adrenalectomized (OVX-ADX) rats increased glucocorticoid (GC) binding in brain regions. The experimental protocol was extended to the spinal cord, a GC target tissue in which ornithine decarboxylase (ODC) is markedly induced by GC treatment. First, we measured GC binding to type I and type II receptors in ventral horn, dorsal horn and lateral funiculus of OVX-ADX rats treated during 4 days with E2 or vehicle. In E2-treated rats, type II receptors increased solely in dorsal horn, whereas type I sites remained unchanged. Second, in a group of OVX-ADX rats receiving dexamethasone (DEX), pretreatment with E2 superinduced ODC in ventral horn and lateral funiculus, but not in dorsal horn. Third, we found that the dorsal horn was relatively enriched in E2 receptors compared to other areas. Therefore, E2 stimulation of GC binding to type II sites may be mediated through E2 receptors localized in the dorsal horn. We suggest that combined treatment with E2 and DEX employs a transsynaptic mechanism for ODC induction at the ventral horn and lateral funiculus, with hormonal interaction taking place at the dorsal horn level. PMID- 8429761 TI - The English medical witness--why so late? PMID- 8429762 TI - Matters forensic. PMID- 8429763 TI - Suicide among AIDS patients. AB - I studied 25 cases of suicide in people diagnosed as having the acquired immunodeficiency syndrome (AIDS) or who thought that they had that disorder, during 1985-1989. Case reports are presented. A discussion ensues concerning the occurrence of this phenomenon and the medical examiner's role in AIDS' public health situation in general. PMID- 8429764 TI - Alcohol-related offending in male special hospital patients. AB - In a sample of 61 male Special Hospital patients, 11 (18%) were identified as having alcohol-related problems prior to admission. Ten of these alcohol-abusers were interviewed to gather further information about their drinking, offending, and the relationship between the two. Alcohol abusers showed more serious criminality than non-abusers, having significantly more previous convictions and being responsible for a disproportionately high number of offences of murder and manslaughter. Alcohol-related problems appear to have developed quickly in adolescence culminating in the index offence at around 30 years of age. At this time, the mean self-reported weekly alcohol consumption was 240 units (one unit = 8.5gm alcohol). All but one man admitted to a relationship between alcohol and crime, and most showed motivation to change their future drinking behaviour. PMID- 8429765 TI - The application of control charts in breath alcohol measurement systems. AB - Measurement provides numerical information, usually to assist in some decision process. Quality control is fundamental to the measurement process if the results are to provide confidence to the decision maker. The degree of quality control required depends on the context and purpose of the measurements. Quality control is particularly important in the forensic measurement of breath alcohol in light of the significant consequences involved. Control charts are an important and widely used tool in quality control for both measuring and manufacturing processes. They help evaluate measurement variability and provide a visual assessment of the system's state of statistical control. Control charts can be developed and applied in a variety of different ways. Several examples are illustrated that apply control charts to breath alcohol measuring systems. Application of these methods should result in improved process monitoring in addition to improved confidence for forensic purposes. PMID- 8429766 TI - A forensic dental identification system with error tolerant algorithms and a review of the prevalence of errors occurring in dental records. AB - After a major disaster, identification of the victims may have to be performed by comparison of the antemortem and post-mortem dental records. Ante-mortem records often contain errors and these must be identified, and compensated for in some way. A computer system for the comparison of large numbers of ante-mortem and post-mortem dental records has been developed, using algorithms which are able to compensate for errors in the records. Data has been collected on the prevalence of errors in dental charts. These were found in 45% of a sample of 50 records. Since matches found by the system may be due to coincidence, a score is computed for each record based on its 'uniqueness'. This allows a confidence limit to be placed on any matches found between the antemortem and post-mortem records. PMID- 8429767 TI - Patients' complaints: the disciplinary system regarding evaluation of physicians' conduct. AB - The study comprises all complaints which patients have made to the Danish National Board of Health in 1984-1985. The Board had disciplinary powers over physicians and advises the public prosecutor in cases of gross negligence that are to be brought before a court. There were 531 complaints on average per year. The frequency of complaints was 1.0 complaint per 10,000 population. A large part of the complaints concerned cases where no physical harm occurred (45%), and in only a small portion (16%) were there serious injuries. The Board found that negligence had occurred in 21% of the complaints, and 1% of the cases were considered to be gross negligence. There were appeals against 17% of the rulings, and of these 9% were either wholly or partly upheld. PMID- 8429769 TI - Underlying patterns of practice in a regional forensic psychiatric service. AB - Using a sample of 306 consecutive referrals, the interactions were investigated between factors relating to patients, referral agencies, assessments and courts. Both the variations in referral rates of diagnoses and the psychiatric histories indicated that the service was being used largely for second opinions on the mentally ill, but for those with neurotic disorder or exacerbations of personality disorder it was more akin to a general psychiatric service. Those thought to pose a sexual risk to adults were particularly likely to have histories of violence, but the opposite was true for those considered a sexual risk to children. Reports initiated by the court are more likely to contain a treatment recommendation and are also more likely to be implemented than those in defence reports. Both the likelihood of a treatment recommendation and its acceptance by the court increased with the severity of diagnosis, with psychiatrist and court reaching full concordance in cases of mental illness. As many as 20 per cent of those seen in prison had mental illness and this was nearly seven times the out-patient rate. Forensic outpatient work was found to be independent of the provision of secure care. These and other findings are examined to help explore the nature of the service. PMID- 8429768 TI - Mentally abnormal homicide--a review of a special hospital male population. AB - The clinical, criminological and demographic characteristics of a Special Hospital population of male patients convicted of homicide are described. The results confirm much previous work on mentally abnormal homicide. Typically the patients come from a disadvantaged social background with poor employment records, a prior history of aggressive behaviour and a diagnosis of schizophrenia. Victims were usually known to the patient and often came from within his circle of family and friends. Only 25% of the patients were receiving treatment at the time of the offence. These findings are discussed in the light of the current literature. PMID- 8429770 TI - Is psychopathic disorder a treatable condition? AB - This article extends upon material presented in a previous issue of this journal (Med. Sci. Law, 1990, 30, 39-44). Three groups of respondents were asked to comment upon the degree to which they considered psychopathic disorder to be a treatable condition. The modest findings suggest that although few clear cut views exist as to the best treatment modality there are firmer indications as to those thought to be unhelpful. PMID- 8429771 TI - An autopsy case of suicide by hanging with multiple stab wounds of the neck and chest. AB - We report, from the aspect of 'acting' capability, an autopsy case of suicide by hanging in which multiple deep stab wounds were observed in the neck and chest. A 49-year-old man was found dead in a state typical of hanging. His car was found about 100m away with a blood-stained climber's knife on the seat. Multiple incised and stab wounds were in the neck, with a lesion of the right inner jugular vein. Three penetrating stab wounds of the chest punctured the lungs, with pooling of about 500ml of blood in the left thorax and about 100ml in the right. Two other shallow stab wounds were in the chest. All the above wounds were explainable as self-inflicted. From the autopsy findings and the circumstantial evidence, it was concluded that he walked approximately 100m after stabbing his neck and chest and finally committed suicide by hanging. PMID- 8429772 TI - Internal carotid artery thrombosis following manual strangulation. AB - Thrombosis of the internal carotid artery is an unusual complication of neck compression, and even more infrequently observed in manual strangulation. It is important in such cases to exclude spontaneous thrombosis which may result from atherosclerotic narrowing of the vessel. PMID- 8429773 TI - Plastic bag suffocation. AB - Suffocation by plastic bag is not common. The finding of a body at the scene of death with a plastic bag in situ could be diagnostically valuable to the pathologist. Otherwise the pathologist may be hard put to arrive at a firm opinion as to how the deceased met his/her death; in fact, in three of the cases reported in this paper, the local pathologist could not give a cause of death when the initial autopsy was performed. Plastic bags could be accidentally placed over the head; put there to help in the inhalation of volatile substances; with homicidal intention; or simply as an effective suicidal method. The object of this paper is to study the cases where bodies were found with a plastic bag over the head, and in particular the cases where the cause of death was due to plastic bag suffocation. PMID- 8429774 TI - Evaluation of physical evidence in a burn case. AB - A young lady was alleged to have died due to accidental burns from a kerosene pressure stove. The dying declaration read that the deceased caught fire accidentally. Evaluation of physical evidence present at the site of the incident, the dying declaration and examination of the body refuted accidental burning from the stove. It proved to be a case of suicide with assistance from the accused due to delivery of insufficient dowry. The accused was convicted on the basis of the physical evidence collected from the scene and evaluated by the forensic experts. PMID- 8429775 TI - Condylomata acuminata in pre-pubertal children. AB - The medico-legal significance of anogenital warts as a marker of sexual abuse in children is unclear. This review article presents three case reports and discusses the possible modes of transmission of papilloma virus in children. Although genital warts can indicate sexual abuse, non-sexual transmission may also occur. PMID- 8429776 TI - Trichomonas vaginalis infection in pre-pubertal girls. AB - This short paper discusses the implications of Trichomonas vaginalis infection of the genital tract in children with respect to suspected child abuse. Two case reports are described to illustrate the problems that can arise in the investigation of such cases T. vaginalis is usually transmitted sexually but the potential for non-sexual transmission does exist. PMID- 8429777 TI - A fatal case of neuroleptic malignant syndrome. AB - A fatal case of Neuroleptic Malignant Syndrome (NMS) affecting a middle-aged woman is presented. Most of the signs and symptoms described for NMS were present and death occurred three hours after the onset of hyperpyrexia. Laboratory and postmortem findings were non-specific. The uses and risks of Haloperidol and Phenelzine on a patient with severe liver impairment are considered. Finally, the medico-legal implications in the context of sudden unexpected death are mentioned. PMID- 8429778 TI - Citrate alterations in primary and metastatic human prostatic adenocarcinomas: 1H magnetic resonance spectroscopy and biochemical study. AB - The objectives of this study were to quantitatively verify the low levels of citrate previously observed in primary human prostatic adenocarcinomas and to determine whether citrate was further reduced in metastatic prostatic cancer. This was accomplished by comparison of citrate concentrations of DU 145 xenografts (a poorly differentiated human prostatic adenocarcinoma cell line grown in nude mice) with concentrations in primary human adenocarcinomas. Following in vivo 1H NMR studies of DU 145 xenografts, citrate concentrations of DU 145 xenografts and surgically removed primary prostatic adenocarcinoma tissue were determined by quantitative high resolution 1H NMR and enzymatic assay. The most significant findings of this study were that citrate concentrations in primary human adenocarcinomas (3.74 +/- 0.19 mumol/g wet weight) were significantly lower than those observed for normal and benign hyperplastic (BPH) prostatic tissues. Furthermore there was a further ten-fold reduction of citrate associated with DU 145 xenografts (0.31 +/- 0.028 mumole/g wet weight) compared with primary prostatic cancer. DU 145 xenografts also exhibited higher levels of uridine diphosphosugars and choline containing metabolites relative to primary prostatic adenocarcinomas. These findings support the hypothesis that citrate is low in primary prostatic cancer and further reduced in metastatic disease. PMID- 8429779 TI - Comparison of localized proton NMR signals of skeletal muscle and fat tissue in vivo: two lipid compartments in muscle tissue. AB - In vivo 1H NMR spectra of small volumes-of-interest (VOI) were localized in human soleus muscle (8 ml) and compared with volume selective spectra of subcutaneous fat tissue and femoral yellow bone marrow (2 ml). All examinations were performed by the double spin echo (PRESS) localization technique. To provide comparability, spectra of different tissues were recorded using identical sequence timing. Clearly improved resolution of the lipid signals of muscle tissue was obtained using long echo times TE > 200 ms. The spectra of muscle tissue exhibit lipid signals that stem from two compartments with a difference of their resonance frequencies of about 0.2 ppm (Larmor frequency difference 12-13 Hz at 1.5 T). The existence of two fatty acid compartments is supported by measurements of the relaxation times and line shape analysis. Both compartments contain fatty acids or triglycerides with similar composition. Probably one compartment corresponds to fat cells within muscle tissue, the other compartment with lower Larmor frequency is located within muscle cells. PMID- 8429780 TI - Carbon-13 "magic-angle" sample-spinning nuclear magnetic resonance studies of human myelin, and model membrane systems. AB - We have obtained high-field (11.7 Tesla), high-resolution carbon-13 solid-state "magic-angle" sample-spinning nuclear magnetic resonance (NMR) spectra of a variety of phospholipids, sphingolipids, myelin and white matter samples, resolving and assigning over 40 resonances in the spectra of human and bovine myelin. The NMR results indicated no large spectral changes due to sample preparation, sample freezing, or brain location, and also no changes in myelin structure detectable via light microscopy, electron microscopy, thin layer chromatography, or sodium dodecyl sulfate-polyacrylamide gel electrophoresis, attributable to the sometimes lengthy NMR data acquisition process. Human myelin and white matter chemical shift assignments were made based on 13C "magic angle" sample spinning (MAS) NMR spectra of individual model lipids, as well as on spectra of lipid mixtures. In all myelin samples there were essentially no features attributable to membrane proteins, with the exception of one small feature due to C zeta of Arg residues, primarily in the myelin basic proteins. The general similarity between the model lipid and intact myelin spectra suggested no major effects of protein on lipid mobility. We have also investigated human myelin samples as a function of developmental age (4, 15, 48 months and adult), and our results showed only small changes in overall lipid composition, although there were significant decreases in lipid hydrocarbon chain unsaturation with age, as determined by computer line-shape simulations of myelin and model compounds. The spectrum of an infant leukoencephalopathy myelin showed marked decreases in galactocerebrosides. Overall, the ability to resolve and assign over 40 resonances in the 13C MAS NMR spectra of myelin, and to detect changes as a function of development and disease, should provide a useful starting point for further more detailed studies of myelin membrane molecular motions, and function. PMID- 8429781 TI - Use of 19F magnetic resonance imaging to measure local cerebral blood volume. PMID- 8429782 TI - A new perfluorocarbon for use in fluorine-19 magnetic resonance imaging and spectroscopy. AB - A new perfluorocarbon, PTBD (perfluoro-2,2,2',2'-tetramethyl-4,4'-bis(1,3 dioxolane)), is described for use in 19F MR imaging and spectroscopy. Two-thirds of the molecular fluorine in PTBD resonates at a single frequency and can be imaged without the use of frequency-selective spin-echo (SE) MRI pulse sequences to suppress chemical shift artifacts. The absence of strong homonuclear spin-spin coupling to the imagable -CF3 groups in PTBD minimizes signal attenuation in 19F SE MRI due to J-modulation effects. For equimolar concentrations of perfluorocarbon, PTBD gives an approximately 17% increase in sensitivity, relative to literature results for perfluorinated amines, at short values of TE (approximately 10 ms) in 19F SE MRI. These attributes allow 19F MRI of PTBD to be performed on standard clinical imaging instrumentation (without special hardware and/or software modification) and an in vivo example in a mouse is shown. This investigation involved characterizing the MR T1 and T2 relaxation times of PTBD as well as the MR spin-lattice relaxation rate, R1 (1/T1), of PTBD as a function of dissolved oxygen concentration. The T1 and T2 relaxation times and R1 relaxation rates of perfluorooctyl bromide (PFOB) were also obtained, under similar experimental conditions, to compare and contrast PTBD with a representative perfluorocarbon that has been widely employed for 19F MRI/MRS applications. PMID- 8429783 TI - Gel-entrapment of perfluorocarbons: a fluorine-19 NMR spectroscopic method for monitoring oxygen concentration in cell perfusion systems. AB - Oxygenation is a major determinant of the physiological state of cultured cells. 19F NMR can be used to determine the oxygen concentration available to cells immobilized in a gel matrix by measuring the relaxation rate (1/T1) of perfluorocarbons (PFC) incorporated into the gel matrix. In calcium alginate gel beads without cells the relaxation rate (1/T1) of the trifluoromethyl group of perfluorotripropylamine (FTPA) varies linearly with oxygen concentration, with a slope of 1.26 +/- 0.15 x 10(-3) s-1 microM-1 and an intercept of 0.50 +/- 0.04 s 1. During perfusion with medium equilibrated with 95%/5% O2/CO2, changes in PFC T1s indicate that the average oxygen concentration was reduced from 894 +/- 102 microM in the absence of cells to 476 +/- 65 microM and 475 +/- 50 microM in the presence of 0.7 x 10(8) EMT6/Ro and RIF-1 murine tumor cells per milliliter of gel, respectively. The presence of 0.2 microliters of FTPA/ml of gel had no effect on the energy status of the cells as indicated by 31P NMR spectra. To calculate oxygen gradients within the beads from the average PFC T1 of the sample, a mathematical model was used assuming that oxygen is the limiting nutrient for cell metabolism and that the cellular oxygen consumption rate is independent of oxygen concentration. Data for EMT6/Ro cells were fit using experimentally determined perfusion parameters together with literature values for cell volume and oxygen consumption rate.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8429784 TI - The sensitivity of magnetic resonance image signals of a rat brain to changes in the cerebral venous blood oxygenation. AB - The sensitivity of magnetic resonance image signals of the brain to the change in the cerebral blood oxygenation was measured in gradient echo images of rat brains at a field strength of 7 T. The sensitivity depended on the blood vessel volume relative to the tissue volume within the image voxel, and signal intensities in the cortical area were well correlated with the change in the venous blood de oxygenation level at the sagittal sinus. Tissue signals in the image (15 ms echo time) showed a sensitivity of 10-20% change for the full range of deoxygenation level from 0-100%. From these observations and image simulations, the extent of the signal response to some neuro-stimulation which induces an increase in regional cerebral blood flow has been estimated for 4 T field strength. PMID- 8429785 TI - Analysis of water-macromolecule proton magnetization transfer in articular cartilage. AB - These studies were designed to establish which structural elements of cartilage are responsible for proton magnetization transfer between water (Hf) and macromolecules (Hr) observed in MRI studies on articular cartilage. Saturation transfer techniques were used to monitor magnetization transfer in vitro on samples of the two major constituents of cartilage: collagen and proteoglycan. Articular cartilage samples were also evaluated in vitro before and after the removal of the proteoglycan fraction. Isolated hydrated collagen exhibited a significant proton magnetization transfer rate with water. In contrast, proteoglycans exhibited no proton magnetization transfer. Articular cartilage, in vitro, exhibited a high degree of magnetization transfer with water protons consistent with previous MRI studies in vivo. Enzymatic removal of proteoglycan from the cartilage did not alter the magnetization transfer rate between Hr and Hf. These data demonstrate that the structure and concentration of the collagen matrix are the predominant determinants of the magnetization transfer process in articular cartilage with little or no contribution from proteoglycans. This specificity of the magnetization transfer effect may prove useful in the noninvasive evaluation of cartilage composition and structure in vivo. PMID- 8429786 TI - Quantitative velocity images from thick slab 2D phase contrast. AB - Thick slab two-dimensional phase contrast (2D PC) angiography provides a large amount of anatomical information in a short acquisition time. Quantitative velocity information is, however, destroyed by the necessary projection dephasing gradient. We present a 2D PC acquisition scheme which retains quantitative velocity information in a thick slab acquisition. A thick slab acquisition produces an image which is a projection through an entire vessel. Accurate velocity measurements must take into account the intravoxel phase cancellation caused by a distribution of velocities within the vessel. In addition to the description of the data acquisition scheme, we outline a method for determining the mean velocity within a vessel having a laminar flow distribution. It is shown that without the consideration for intravoxel phase cancellation, the mean velocity measurement from thick slab images overestimates the true mean velocity by 10%. PMID- 8429787 TI - Fast perfluorocarbon imaging using 19F U-FLARE. AB - The application of an ultra-fast low angle RARE technique for the 19F imaging of perfluorocarbons (PFCs) used as temporary blood substitutes is described. This sequence is attractive for fast 19F imaging studies that measure the biodistribution of PFCs in vivo, due to its high signal-to-noise ratio. Extensions of this technique for the chemical shift selective measurement of fluorine T1 values are presented. Using the linear dependence between the oxygen partial pressure (pO2) and the T1 relaxation rate of PFC resonances this technique makes possible the fast in vivo measurement of oxygen tension. Using the sequence in a diffusion sensitized form 19F measurements of the diffusion constants of PFCs are also presented. Phantom experiments to test the methods, and in vivo images obtained in rat studies are given and discussed. PMID- 8429788 TI - The diffusion sensitivity of fast steady-state free precession imaging. AB - Steady-state free precession (SSFP) imaging with an added field gradient pulse is strongly sensitive to self-diffusion and other motions of water. In an earlier theoretical analysis of diffusion attenuation due to a single gradient pulse Wu and Buxton (J. Magn. Reson. 90, 243, 1990) concluded that the diffusion sensitivity would be increased with smaller flip angles. In this paper a partial partition analysis of the different echo pathways contributing to the signal is used to illustrate the contribution of stimulated echo pathways with long diffusion times as the source of the enhanced diffusion sensitivity with low flip angles. Experimental imaging studies in phantoms and the brain of a human subject demonstrate substantially greater signal attenuation with small flip angles (< 30 degrees). The theoretical equation of Wu and Buxton provides a reasonable fit to the experimental data, accounting for the flip angle and TR dependence, but the estimated diffusion coefficients are larger than expected from previous studies. The large attenuation observed in the human studies, particularly in cerebrospinal fluid, is most likely due to other tissue motions. Both the theoretical calculations and the experimental data show that for the same gradient strength the diffusion sensitivity of SSFP is much greater than the diffusion sensitivity of conventional spin-echo methods. PMID- 8429789 TI - Effect of glucose and confluency on phosphorus metabolites of perfused human prostatic adenocarcinoma cells as determined by 31P MRS. AB - A series of perfused cell 31P MRS studies were conducted using a well established human prostate adenocarcinoma cell line (DU 145) at different phases of growth, and exposed to varying glucose concentrations during growth. The spectral characteristics of perfused DU 145 cells were compared with the same cells grown in nude mice (xenografts). Perfused DU 145 cells had lower levels of inorganic phosphate and phosphocreative relative to in vivo nude mice xenografts. 31P MR spectra obtained from perfused cells at different phases of growth and exposed to varying glucose concentrations during grown suggest that increases in diphosphodiester levels are associated with high glucose concentrations and confluency. Perfused DU 145 cells grown in 5.5 mM glucose and harvested at log phase of growth best reflected the phosphorus MR spectra of the same cell line grown in nude mice. PMID- 8429790 TI - pH mapping in living tissues: an application of in vivo 31P NMR chemical shift imaging. AB - Chemical shifts were extracted from in vivo 3-dimensional 31P NMR CSI data and pH images were constructed. The images could spatially resolve tissue pH ranging from 5.8 to 7.2 (with uncertainty of 0.11-0.17 pH unit) in an ischemia reperfusion model of diabetic rat calf muscles. PMID- 8429791 TI - Specific in vitro labeling of cells with a fluorine-19 probe encapsulated in antibody-targeted liposomes: a F-19 NMR spectroscopy study. AB - Liposomes containing dexamethasone phosphate (DMp) were covalently coupled to protein A and then incubated with murine L929 fibroblast and RDM4 thymoma cells in the presence of monoclonal antibodies specific for the major histocompatibility complex. The detection of the specific F-19 labeling of cells is rapid (minutes). Such a strategy might be useful to study the kinetics of internalization processes of bound liposomes on cultured living cells. PMID- 8429792 TI - Reinvestigation of the transmembrane difference in 7Li NMR T1 values in Li(+) loaded human erythrocyte suspensions. AB - In contrast to the findings in a recent study (R. P. Gullapalli, R. M. Hawk, R. A. Komoroski, Magn. Reson. Med. 20, 240 (1991)), we found that, even at high hematocrits, the T1 values for extracellular 7Li were at least three times longer than those for intracellular Li+. We conclude that a transmembrane difference in T1 values can be used for separate observation of intracellular and extracellular Li+ in human red blood cell suspensions. PMID- 8429793 TI - On self-refocusing pulses for rapid gradient-echo imaging. AB - Self-refocusing slice selective radio frequency pulses could potentially be used to reduce imaging times in rapid gradient-echo sequences. However, in practice, the improvement is likely to be no greater than about 20%, and at the expense of much increased radio frequency power requirements. PMID- 8429794 TI - Equivalent circuit for birdcage resonators. AB - We present an equivalent circuit analysis for both low pass and high pass birdcage resonators loaded with lossy samples. In a generalization of the method of Hoult and Lauterbur (J. Magn. Reson. 34, 425 (1979)), we also derive circuit component values by application of the laws of electrodynamics. Measured resonance spectra, quality factors, and feed point impedances in a test resonator are shown to be in agreement with those predicted by the proposed model. PMID- 8429795 TI - Aortic ghost artifact in ultrashort TE multislice gradient echo MR images is not increased by paramagnetic enhancement. AB - Pulsation artifact on gradient echo images with ultrashort TE (i.e., < 3 msec) and intermediate TR is primarily from view-to-view amplitude modulation. Paramagnetic contrast agents increase the signal from blood during diastole without increasing the intensity of unsaturated systolic blood, decreasing signal modulation between systole and diastole. In a phantom and in humans, artifact decreased or remained the same following contrast enhancement. PMID- 8429796 TI - Endogenous susceptibility contrast in myocardium during apnea measured using gradient recalled echo planar imaging. AB - Gradient recalled echo planar imaging was used to monitor changes in myocardial and left ventricular chamber blood intensity during apnea in rats. Significant signal loss in both blood (to 62 +/- 5% and 51 +/- 6% of baseline) and myocardium (to 79 +/- 2% and 76 +/- 3% of baseline) was observed at 45 and 90 s apnea while O2 saturation decreased from 98 +/- 1% to 62 +/- 7% and 36 +/- 9%, respectively. These results show that myocardial intensity is modulated by alterations in blood oxygenation. PMID- 8429797 TI - Functional mapping of the human visual cortex at 4 and 1.5 tesla using deoxygenation contrast EPI. AB - The effects of photic stimulation on the visual cortex of human brain were studied by means of gradient-echo echo-planar imaging (EPI). Whole-body 4 and 1.5 T MRI systems, equipped with a small z axis head gradient coil, were used. Variations of image intensity of up to 28% at 4 T, and up to 7% at 1.5 T, were observed in primary visual cortex, corresponding to an increase of blood oxygenation in regions of increased neural activity. The larger effects at 4 T are due to the increased importance of the susceptibility difference between deoxygenated and oxygenated blood at high fields. PMID- 8429798 TI - QUEST--a quick echo split NMR imaging technique. AB - A novel millisecond NMR imaging method is introduced, which generalizes the principle of stopped-pulse experiments. It has been dubbed QUEST for QUick Echo Split imaging Technique. Repeated dephasing and excitation is used to divide a primary free induction decay into an exponentially growing number of echoes. Very few gradient lobes and RF pulses already generate large numbers of echoes, so neither gradient switching speed nor RF absorption represents any real limitation. True spin echoes are easily produced. PMID- 8429799 TI - A novel oestradiol--desogestrel preparation for hormone replacement therapy: effects on hormones, lipids, bone, climacteric symptoms and endometrium. AB - Desogestrel is a strong progestogen with low androgenicity which has so far been used only in oral contraceptives. We studied the feasibility of administering desogestrel in combination with oestradiol as hormone replacement therapy (HRT). Thirty women received a sequential combination containing 1.5 mg micronised oestradiol (24 days) and 0.15 mg desogestrel (last 12 days of cycle) for 6 months. At that stage 6 of the women dropped out; the remaining 24 were studied for a total of 12 months. The treatment alleviated vasomotor symptoms effectively in all the women and induced regular withdrawal bleeding in 86% of them. Secretory changes were observed in the endometria of 16 of the 20 women with adequate endometrial samples assessed after 12 months of treatment. No signs of hyperplasia or atypia were found. Six months of treatment resulted in a decrease in the mean serum follicle-stimulating-hormone concentration from 66.2 (+/- 4.3, S.E.M.) to 23.3 (+/- 3.1) IU/l and a rise in the oestradiol and sex-hormone binding globulin concentrations from 87.9 (+/- 13.7) to 233.1 (+/- 20.4) pmol/l and from 52.1 (+/- 4.6) to 70.2 (+/- 5.6) nmol/l, respectively. Testosterone levels decreased. There were significant reductions in serum total and low density lipoprotein (LDL) cholesterol and triglycerides. After 12 months of treatment high-density lipoprotein (HDL) cholesterol values did not differ significantly from the pretreatment levels. The HDL/LDL and HDL/total cholesterol ratios increased. The treatment reduced bone turnover as indicated by decreases in bone alkaline phosphatase and osteocalcin serum levels and by lowered urinary calcium/creatinine and hydroxyproline/creatinine ratios. An increase of about 2% in forearm bone mineral density was also observed. This new oestradiol desogestrel preparation therefore appears to be a promising alternative form of HRT. It alleviates climacteric symptoms effectively, exhibits favourable effects on serum lipids and lipoproteins and prevents bone loss. PMID- 8429800 TI - A two-year double-blind controlled study of the clinical effect of combined and sequential postmenopausal replacement therapy and steroid metabolism during treatment. AB - A total of 151 postmenopausal women were randomly allocated to 3 groups for treatment with hormone replacement therapy. One group received combined therapy (2 mg oestradiol (E2) and 1 mg norethisterone acetate (NETA) daily), the second group was placed on sequential therapy (2 mg E2 for 12 days, 2 mgE2 and 1 mg NETA for 10 days and 1 mg E2 for 6 days), while the third was given placebo. Treatment was administered over 24 cycles of 28 days. The two active treatments were equally effective in relieving climacteric symptoms. In the combined therapy group, 62% of the women experienced spotting and/or breakthrough bleeding during the first 3 cycles; thereafter this proportion decreased to between 3 and 18% in each of the following three-cycle periods. Sixty-four percent (64%) of these women had no more bleeding after the first 3 cycles. Endometrial atrophy was detected in 93% of the women in this group after 24 cycles of therapy. Bleeding irregularities occurred during the first 3 cycles in 27% of the patients treated with sequential therapy and in 21% of those receiving placebo. In the subsequent 3-cycle periods these figures fell to below 10% in the 2 groups. In all 3 groups weight remained stable but blood pressure increased equally in the actively treated groups and the placebo group. The levels of follicle-stimulating hormone (FSH), sex-hormone-binding globulin (SHBG) and the free fraction of E2 in serum were significantly lower in the combined therapy group than in the sequential therapy group. The higher level of free E2 in the latter group may have been caused by a decrease in metabolism associated with the increased SHBG concentration. It was concluded that combined treatment with E2 and NETA might provide an alternative to sequential treatment in postmenopausal women willing to tolerate the initial high risk of breakthrough bleeding/spotting in order to avoid subsequent regular bleeding. In the subgroup of women in whom bleeding irregularities continue, sequential treatment should be considered. PMID- 8429801 TI - Does low-dose, transdermal, norethisterone acetate reliably cause endometrial transformation in postmenopausal oestrogen-users? AB - In a prospective study, the symptomatic, psychological and endometrial effects of a combined oestradiol/progestogen transdermal therapeutic system which delivers a low daily dose of norethisterone acetate have been investigated in 18 postmenopausal women. Treatment was given in 28 day cycles. Patients received transdermal oestradiol 50 micrograms/day continuously and transdermal norethisterone acetate, 0.1-0.15 mg/day, was added for 14 days. Treatment was given for between 5 and 7 cycles. One patient discontinued therapy because of a skin reaction. Low dose, transdermal norethisterone acetate caused few adverse symptomatic or psychological side-effects and appeared well tolerated. Fourteen endometrial samples were obtained during the combined (norethisterone acetate/oestradiol) phase of the 5th, 6th or 7th treatment cycle. None showed hyperplastic or malignant change but proliferative endometrium, indicative of an inadequate progestational stimulus, was observed in 4 biopsy samples. Three of these patients had reported breakthrough bleeding during therapy. For these reasons, we regard this dose of transdermal norethisterone acetate as inadequate in combination with transdermal oestradiol 50 micrograms/day for routine use in postmenopausal oestrogen users. PMID- 8429802 TI - Hypercalciuria and urinary saturation measurements in climacteric women. AB - Fifteen percent (20/130) of a group of climacteric women on diets of their choice had urinary calcium (Ca) levels exceeding 4 mg/kg per day. Most of these hypercalciuric subjects had a daily Ca intake of 0.4-0.5 g. Their bone turnover rates were raised and high Ca absorption was observed in 4 cases. Serum Ca and total protein and glomerular filtration rates were normal in all the hypercalciuric patients. The calcium/creatinine (Ca/Cr) ratio (mg l-1/mg l-1, fasting, 09:00-10:00 h) was measured in 72 climacteric women, 35 of whom (49%) had ratios > 0.1. The latter defines a relative hypercalciuria as compared with premenopausal Ca excretion levels. Only 5 of the 35 subjects had calciuria levels above 4 mg/kg per day. The Ca/Cr ratio cannot replace daily urinary Ca measurements for the screening of subjects in whom calciuria may exceed net Ca absorption. Urinary saturation measurements were carried out in 70 women. Supersaturation was observed only in the case of Ca oxalate (CaOx) among several calcium salts usually found in urinary stones. CaOx supersaturation was observed in 95% of the hypercalciuric subjects and in 48% of the rest of the women investigated. The relatively high frequency of CaOx supersaturation can be attributed in part to the decreased excretion of citrate associated with ovarian failure. Oestrogen replacement therapy increased citrate excretion and lowered the level of CaOx supersaturation. Ca supplementation (1 g Ca/day) reduced the degree of supersaturation as a result of the concurrent reduction in oxalate excretion. PMID- 8429803 TI - Sexual desire and menopausal development. A prospective study of Danish women born in 1936. AB - This study examines the nature of sexual desire experienced during the climacteric and attempts to identify predictors for frequency and change of sexual desire using a prospective design. Four-hundred seventy-four women constituting 76% of the original cohort of a general population of Danish women born in 1936 were examined by interview and questionnaire at the ages of 40, 45 and 51. The main items were sexual behaviour, expectations towards or experience of menopause, general health status and social background. Most women (70%) experienced no change in sexual desire during the study period. The 51-year-old women's experience of frequency or change in sexual desire was not related to menopausal status. Frequency of sexual desire was highly correlated to previous and present subjective health status, former sexual activity, partner availability and social status. Anticipations of declining sexual desire as a consequence of menopause actually predicted a decrease. Finally, decrease in sexual desire correlated significantly with the women's subjective assessment of being climacteric. PMID- 8429804 TI - Characteristics of menopause in three socioeconomic urban groups in Karachi, Pakistan. AB - To determine age, symptoms and treatment choices in Pakistani women with spontaneous menopause, three groups in Karachi were interviewed using a structured questionnaire. The groups consisted of 250 poor slum dwellers, 250 middle class clinic attenders and 150 wives of retired military officers. Interviews were carried out in 1989 and 1990 by members of a team from the Aga Khan University consisting of a gynaecologist, a community health physician and two senior medical students. All interviewees had natural menopause at least 12 months previously. The results showed a mean age of 47 years for menopause in all groups. One in five women were symptomatic in the poorest group but one in two were in the other groups. Treatment was sought by 6% in the poor group, 26% of the middle class group and 38% of the most privileged group. Age at menopause was 4 years earlier than in most reports and fewer women had menopausal symptoms. These urban women may not represent the situation across the country but, as life expectancy in Pakistan is rising (now 56.4 years for women), menopausal problems may increase. PMID- 8429805 TI - Setting up a specialist menopause clinic in Ireland: the initial experience. AB - The Rotunda Hospital Dublin set up a Menopause Clinic in February 1990. Eighty one new patients were seen in the first 6 months of operation. Most presented either with symptoms attributable to oestrogen deficiency (36%) or non-specific symptoms associated with depression or ageing (26%). There was a high incidence of significant family/social/martial problems (25%) and abnormal vaginal bleeding (20%). The case is made for such specialised clinics; expertise with respect to hormone replacement therapy and its manipulation develops, problems of specific socio-cultural settings can be identified, and screening programmes for disease coordinated. PMID- 8429806 TI - Emergency public health surveillance in response to food and energy shortages- Armenia, 1992. AB - Living conditions in Armenia have deteriorated since 1988 as a result of an economic blockade related to a territorial conflict between Armenia and a neighboring country. The effects of this blockade--a drastic reduction in available food, heating fuel, gasoline, electricity, health services, drugs, and vaccines--have placed residents of Armenia at increased risk for morbidity and mortality from nutritional deficiencies, infectious diseases, and hypothermia. To assess and monitor the current health and nutritional status of residents of Armenia, the Armenian National Institute of Health, the U.S. Agency for International Development (USAID), and CDC have developed the Emergency Public Health Information Surveillance System (EPHISS). This report summarizes preliminary results for 1992. PMID- 8429807 TI - Capnocytophaga canimorsus sepsis misdiagnosed as plague--New Mexico, 1992. AB - Since 1961, 200 human isolates of Capnocytophaga canimorsus--a gram-negative bacterium--have been sent to CDC for identification. Infections with this organism may result in a spectrum of manifestations ranging from self-limiting cellulitis to fatal septicemia; most fatal infections have occurred in persons with a history of asplenia, alcoholism, or hematologic malignancy. In most (77%) cases, infection is preceded by a bite or other exposure to dogs. This report summarizes the investigation by the New Mexico Health and Environment departments and CDC of a fatal case of C. canimorsus infection in a resident of New Mexico. This case was initially misdiagnosed as human plague. PMID- 8429808 TI - Approaches to improving adherence to antituberculosis therapy--South Carolina and New York, 1986-1991. AB - Patients with tuberculosis (TB) who fail to complete a standard course of antituberculosis (anti-TB) therapy are at increased risk for treatment failure and may play a role in both the emergence of drug-resistant strains of Mycobacterium tuberculosis and further spread of TB. During 1986-1991, the South Carolina Department of Health and Environmental Control and the New York City Department of Health (NYCDH) attempted to improve patient adherence to anti-TB therapy by employing a combination of strategies that included incentives, directly observed therapy (DOT) (i.e., health-care worker observation of the patient ingesting each dose of medication), court-ordered DOT, and commitment for inpatient management. This report describes the experiences of selected strategies in South Carolina and New York City and provides recommendations for improving patient adherence to anti-TB therapy. PMID- 8429809 TI - Measles--Duval County, Florida, 1991-1992. AB - An outbreak of measles occurred in northeastern Florida (Bradford, Clay, Duval, Nassau, Putnam, and St. Johns counties) in 1991 and early 1992. A total of 193 confirmed cases of measles were reported to the Florida Department of Health and Rehabilitative Services (FDHRS). This report summarizes an investigation of the outbreak in Duval County (1990 population: 676,556), which reported 146 (76%) of the cases. PMID- 8429810 TI - Adult blood lead epidemiology and surveillance--United States, third quarter, 1992. AB - In September 1992, CDC's National Institute for Occupational Safety and Health (NIOSH) began quarterly reporting of adult elevated blood lead level (BLL) data from state-based surveillance programs. To support these efforts, NIOSH has established the Adult Blood Lead Epidemiology and Surveillance (ABLES) program. PMID- 8429811 TI - Preliminary report: foodborne outbreak of Escherichia coli O157:H7 infections from hamburgers--western United States, 1993. PMID- 8429812 TI - Update: availability of sulfadiazine--United States. AB - Sulfadiazine is a sulfa drug commonly used in combination with pyrimethamine to treat toxoplasmosis in patients with acquired immunodeficiency syndrome (AIDS) and newborns with congenital infections. In December 1992, CDC reported that the domestic manufacturer of sulfadiazine had ceased production in October 1992 and that no large inventories of the drug were available from major distributors (1). On February 8, CDC obtained permission from the Food and Drug Administration (FDA) to proceed with an "Investigational New Drug" (IND) application protocol under which sulfadiazine may be imported and made available to physicians until a domestic commercial source is re-established. PMID- 8429813 TI - Isolation of Vibrio cholerae O1 from oysters--Mobile Bay, 1991-1992. AB - On July 2, 1991, during routine monitoring, the Food and Drug Administration (FDA) isolated toxigenic Vibrio cholerae O1, serotype Inaba, biotype EI Tor from oysters and intestinal contents of an oyster-eating fish taken from closed oyster beds in Mobile Bay. This isolate was indistinguishable from the Latin American epidemic strain and differed from the strain of V. cholerae O1 that is endemic to the Gulf Coast. This report summarizes the public health response to this isolation of V. cholerae O1. PMID- 8429814 TI - Update: dracunculiasis eradication--Ghana, 1992. AB - The reported incidence of dracunculiasis (i.e., Guinea worm disease) in Ghana declined substantially during 1992--the third consecutive year in which reports of known cases declined. This report summarizes 1992 surveillance data for Ghana that are being used to monitor progress toward eradication of this disease. PMID- 8429815 TI - Business responds to AIDS program--December 1992-February 1993. AB - Of all acquired immunodeficiency syndrome (AIDS) cases reported through 1991 in the United States, 76% occurred among persons aged 25-44 years; persons aged 15 44 years comprise approximately 50% of the United States workforce. On December 1, 1992, CDC introduced "Business Responds to AIDS" (BRTA)--a new program for the primary prevention of human immunodeficiency virus (HIV)/AIDS. BRTA, which was introduced by satellite teleconference, encourages business and labor leaders in U.S. communities to develop comprehensive workplace HIV-prevention education programs (3). This report describes BRTA and summarizes the response--from December 1, 1992, through February 5, 1993--to the launch of the national program. PMID- 8429816 TI - Unintentional carbon monoxide poisoning following a winter storm--Washington, January 1993. AB - Carbon monoxide (CO) poisoning was a major health consequence of a severe storm that struck the Puget Sound region of western Washington state the morning of January 20, 1993. Wind gusts up to 94 miles per hour interrupted electrical power for an estimated 776,000 residents, and during the 4 nights following the storm, temperatures fell to near freezing. Because of the use of alternative sources of energy for indoor cooking and home heating, the risk of exposure to CO increased for many persons. This report summarizes cases of storm-related CO poisoning among persons who were initially evaluated at Seattle's Harborview Medical Center (HMC) or who were referred to the Virginia Mason Medical Center (VMMC) for hyperbaric oxygen therapy. PMID- 8429817 TI - Toddler deaths resulting from ingestion of iron supplements--Los Angeles, 1992 1993. AB - During June 1992-January 1993, five children aged 11-18 months in the Los Angeles area died after ingesting iron supplement tablets. The first death was reported by the Consumer Product Safety Commission (CPSC) to the Los Angeles County Department of Health Services (LADHS) in November 1992 and then reviewed by the Los Angeles County Child Death Review Committee. On January 6, the Los Angeles County coroner's office reported three additional deaths associated with ingestion of iron to the Child Death Review Committee and the health department. A fifth death resulting from iron poisoning was reported January 29. This report summarizes preliminary information from an investigation of these cases. PMID- 8429818 TI - Prenatal care and pregnancies complicated by diabetes--U.S. reporting areas, 1989. AB - Women who are pregnant and who have diabetes (either established [i.e., diabetes mellitus diagnosed before conception] or gestational [i.e., carbohydrate intolerance of variable severity with onset or first recognition during pregnancy]) are at increased risk for adverse fetal and maternal outcomes (1,2). To reduce these risks, CDC, the American Diabetes Association, and other health care professionals recommend that women who are of childbearing age and have diabetes undergo prepregnancy counseling and that all pregnant women receive early and continued prenatal care, including screening for gestational diabetes sometime during weeks 24-28 of pregnancy (3,4). Although appropriate prenatal care practices, including screening for gestational diabetes, have been recommended by CDC and others, there is no system to routinely monitor national trends in prenatal practices among mothers with diabetes mellitus. This report summarizes an analysis of U.S. birth certificates in 1989 to characterize racial/ethnic differences in prenatal care for live births, including those among mothers whose pregnancies were complicated by diabetes. PMID- 8429819 TI - Subtype selectivity of a novel endothelin antagonist, FR139317, for the two endothelin receptors in transfected Chinese hamster ovary cells. AB - We investigated the receptor-binding properties and the antagonist activities of FR139317, a novel endothelin (ET) antagonist, in transfected Chinese hamster ovary cells permanently expressing the two ET receptor subtypes (ETA and ETB). In displacement analysis using membrane preparations derived from the receptor expressing cells, FR139317 showed a high affinity for ETA (Ki = 1 nM) and a lower affinity for ETB (Ki = 7.3 microM). FR139317 inhibited ETA-mediated phosphatidylinositol hydrolysis and arachidonic acid release and produced a parallel shift in the dose-response curve for ET-1, with respective pA2 values of 8.2 and 7.7. In contrast, FR139317 had no inhibitory effects on these ET-1 induced responses in ETB-expressing cells. FR139317 itself showed no stimulatory effects on phosphatidylinositol hydrolysis and arachidonic acid release in ETA- and ETB-expressing cells. Thus, FR139317 is a potent, competitive, and highly selective antagonist for ETA. This compound should be a powerful tool for investigation of the physiological properties of ETA and exploration of its role in diseases. PMID- 8429820 TI - Biochemical characterization of a 22-kDa high affinity antiischemic drug-binding polypeptide in the endoplasmic reticulum of guinea pig liver: potential common target for antiischemic drug action. AB - The phenylalkylamine emopamil prevents brain damage due to experimental cerebral ischemia. Stereoselective, high affinity, binding sites for (-)-[3H]emopamil in guinea pig brain cortex and liver membranes have been proposed to mediate its antiischemic effect. Using [N-methyl-3H]LU49888 as a photoaffinity probe we now provide evidence that the cation-sensitive emopamil binding site is localized on a 22-kDa polypeptide in guinea pig liver, kidney, lung, and adrenal gland. This 22-kDa polypeptide binds other antiischemic drugs with high affinity and is a nonglycosylated integral membrane protein of the endoplasmic reticulum. It can be solubilized with digitonin without changes in its drug-binding properties. The solubilized binding activity has a sedimentation coefficient of 12.0 +/- 0.4 S and an apparent Stokes radius of 6.0 +/- 0.1 nm. From these data it is concluded that the 22-kDa polypeptide is associated in a larger oligomeric complex with a molecular mass of at least 84 kDa. [N-methyl-3H]LU49888 also specifically labels a second 27-kDa polypeptide in the endoplasmic reticulum, which can be distinguished from the 22-kDa polypeptide by its pharmacological and hydrodynamic properties. The photolabeled 22-kDa polypeptide was partially purified under denaturating conditions. This will allow the further structural analysis of this putative target for antiischemic drugs. PMID- 8429821 TI - Development of antisera selective for m4 and m5 muscarinic cholinergic receptors: distribution of m4 and m5 receptors in rat brain. AB - A portion of the cDNA sequence corresponding to the third intracellular loop of either the m4 or m5 muscarinic cholinergic receptor was ligated into the pRIT23 or pET-3a expression vector, respectively. The expressed fusion proteins were purified and used to develop selective polyclonal antisera to the m4 and m5 muscarinic receptors. These antisera were used in an immunoprecipitation protocol to examine quantitatively the distribution of receptor subtypes in regions of rat brain. The density of m4 receptors in rat brain increased in the caudal to rostral direction. The highest levels of m4 receptors were detected in the striatum (1280 fmol/mg) and olfactory tubercle (750 fmol/mg). Low levels of m5 receptors were detected in several brain regions (< 25 fmol/mg). By combining the previously determined receptor densities for the m1, m2, and m3 receptors and results obtained with the newly developed antisera to m4 and m5 receptors, it was determined that 86-99% of the [3H]quinuclidinyl benzilate binding sites in several brain regions were immunoprecipitated. In addition to measuring receptor densities in rat brain, the immunoprecipitation protocol was used to quantify muscarinic receptor levels in tissues reported to express mRNA encoding the m4 receptor. Thus, although only m4 mRNA has been detected in rabbit lung, NG108-15 cells, and N1E-115 cells, both rabbit lung and NG108-15 cells possess both m2 (rabbit lung, 27%; NG108-15 cells, 31%) and m4 (rabbit lung, 55%; NG108-15, 42%) receptors, whereas N1E-115 cells were found to have both m1 (15%) and m4 (65%) receptors. These antisera will be useful in studies of receptor regulation and in determining alterations in density that may occur after pharmacological or physiological manipulations and in various disease states. PMID- 8429822 TI - Inhibition by glucocorticoids of the formation of interleukin-1 alpha, interleukin-1 beta, and interleukin-6: mediation by decreased mRNA stability. AB - The mechanism by which glucocorticoids inhibit interleukin (IL)-1 and IL-6 formation in human monocytes and a promonocytic cell line activated by Escherichia coli lipopolysaccharide was analyzed. Dexamethasone (DEX) decreased levels of IL-1 alpha and IL-1 beta mRNAs in a dose-related fashion. The DEX induced decrease in levels of IL-1 alpha and IL-1 beta mRNAs was abolished by the steroid receptor antagonist RU486. The levels of IL-1 alpha and IL-1 beta proteins within the cells and of IL-1 beta in the culture medium were decreased by DEX to comparable extents, so that DEX had no detectable effect on cytokine secretion. DEX did not influence lipopolysaccharide-induced transcription of the IL-1 beta gene in monocytes. However, DEX markedly decreased the stability of IL 1 beta mRNA, as shown both by steady state measurements and by pulse-labeling. DEX-induced instability of IL-1 beta mRNA required protein synthesis. DEX was also found to be a potent inhibitor of IL-1-induced expression of the IL-6 gene in connective tissue-type cells from the synovium of patients with rheumatoid arthritis. Inhibition of the formation of proinflammatory cytokines, including IL 1 beta and tumor necrosis factor-alpha, is a mechanism by which glucocorticoids exert anti-inflammatory effects. Inhibition by glucocorticoids of the expression of IL-1 alpha in antigen-presenting cells could decrease the capacity of the cells to stimulate the proliferation of T lymphocytes. This activity, as well as inhibition of the production and effects of IL-1 beta, including induced formation of IL-6 and of certain lymphokines, could explain the immunosuppressive effects of glucocorticoids. PMID- 8429823 TI - Purification from liver microsomes from untreated cynomolgus monkeys of cytochrome P450 closely related to human cytochrome P450 2B6. AB - A cytochrome P450 (P450) (referred to as P450CMLa) was purified and characterized from hepatic microsomes from untreated cynomolgus monkeys (Macaca irus). The final preparation was electrophoretically homogeneous and its estimated minimum molecular mass was 49.5 kDa. The amino-terminal amino acid sequence of the protein (first 34 residues) closely resembled that of the protein encoded by the 2B6 cDNA from humans (94%). This protein was cross-reactive with antibodies raised against P450 2B1 (P450 b), P450 2B11 (P450 PBD-2), and P450GP-1, which were purified from hepatic microsomes from phenobarbital-pretreated rats, beagle dogs, and guinea pigs, respectively. Also, the antibody raised against P450CMLa was able to cross-react with P450 2B1, P450 2B11, and P450GP-1. P450CMLa was capable of catalyzing benzphetamine N-demethylation and testosterone 16 beta hydroxylation in a reconstituted system. Anti-P450CMLa antibody inhibited the activity of testosterone 16 beta-hydroxylase but not the activities of testosterone 2 beta- and 6 beta-hydroxylases in liver microsomes from cynomolgus monkeys. The content of P450CMLa, as estimated by immunoblot analysis, was 70 pmol/mg (about 5% of total P450). The protein immunoreactive with the anti P450CMLa antibody was also present in liver microsomes from Japanese monkeys, baboons, common marmosets, and common squirrel monkeys. In liver microsomes from common squirrel monkeys, the content of protein immunoreactive with the anti P450CMLa antibody and the activity of testosterone 16 beta-hydroxylase were effectively increased by pretreatment with phenobarbital. The antibody against P450CMLa strongly inhibited the activity of testosterone 16 beta-hydroxylase in liver microsomes not only from untreated cynomolgus monkeys but also from phenobarbital- and pregnenolone 16 alpha-carbonitrile-pretreated common squirrel monkeys. These results indicated that the P450CMLa purified here is very similar to the forms of P450 classified into the 2B subfamily, in its amino-terminal amino acid sequence, catalytic activities, and immunochemical properties. PMID- 8429824 TI - Quinolone antibacterial agents: relationship between structure and in vitro inhibition of the human cytochrome P450 isoform CYP1A2. AB - The inhibitory effect of 44 quinolone antibacterials and derivatives (common structure, 4-oxoquinoline-3-carboxylic acid) on cytochrome P450 isoform CYP1A2 activity was tested using human liver microsomes and caffeine 3-demethylation as a specific test system for this enzyme. By direct comparison of molecules differing structurally in only one position, the following structure-activity relationships were found. 3'-Oxo derivatives had a reduced or similar activity and M1 metabolites (cleavage of piperazinyl substituent) had a greater inhibitory activity, compared with the parent molecule. Alkylation of the 7-piperazinyl substituent resulted in a reduced inhibitory potency. Naphthyridines with an unsubstituted piperazinyl group at position 7 displayed a greater inhibitory potency than did corresponding quinoline derivatives. Derivatives with a fluorine substitution at position 8 had only a minor effect. Molecular modeling studies with inhibitors and caffeine showed that it is possible to explain the potency of the quinolones to inhibit CYP1A2 on a molecular level. The keto group, the carboxylate group, and the core nitrogen at position 1 are likely to be the most important groups for binding to the active site of CYP1A2, because the molecular electrostatic potential of all inhibitors is very similar to that of caffeine in these regions. The presence of a piperazinyl substituent, however, seems to be no prerequisite for inhibitory potency. Finally, an equation to estimate the potency to inhibit CYP1A2 was developed by quantitative structure-activity relationship analysis. PMID- 8429825 TI - Rat small intestinal cytochromes P450 probed by warfarin metabolism. AB - Small intestinal cytochromes P450 (P450s) provide potential first-pass metabolism of ingested xenobiotics. To investigate this system, this study addresses the procedure for elution of enterocytes from male rat small intestine, histological evaluation of the elution procedure, and assessment of the functional microsomal P450s in small intestine of untreated and induced rats, using warfarin metabolism as a probe. Histologically it was demonstrated that villous enterocytes are initially detached in sheets and are subsequently eluted without clear resolution into villous tip, midvillous, and lower villous cells, contrary to previous reports. Crypt cells are eluted after cells from the villus. The following functional P450s were identified, using stereo- and regioselectivity of warfarin metabolism, in small intestinal microsomes: P4502B1 in untreated rats, P4501A1 in beta-naphthoflavone-induced rats, P4502B1 in phenobarbital-induced rats, and P4503A1/2 in pregnenolone-16 alpha-carbonitrile-induced rats. In contrast to hepatic microsomes from untreated or induced rats, P4502C11 and -2C6 were not present or inducible by these inducing agents in rat intestine. Western immunoblots, warfarin assays, and P450 assays all indicated that beta naphthoflavone induced P4501A1 in small intestinal villous and crypt cells, but in contrast to the liver neither apo-P4501A2 nor functional P4501A2 was induced. PMID- 8429826 TI - Correlation of human cytochrome P4502C substrate specificities with primary structure: warfarin as a probe. AB - The regio- and stereoselectivity of warfarin metabolism have been used to assess structure-function relationships of human P4502C subfamily members. Metabolism was investigated using a yeast cDNA expression system in which full length cDNAs for P4502C8, -2C9 (alleles Arg144 Tyr358 Ile359 Gly417 and Arg144 Tyr358 Leu359 Gly417), -2C18 (alleles Thr385 and Met385), and -2C19 were expressed. Additionally, two mutations reported in other P4502C9/2C10 alleles were individually introduced into P4502C9 by site-directed mutagenesis, to yield Cys144 Tyr358 Ile359 Gly417, Arg144 Tyr358 Ile359 Asp417, and Arg144 Cys358 Ile359 Gly417, which were expressed in yeast; their ability to metabolize warfarin was then studied. Warfarin metabolism by purified preparations of P4502C9 allele Arg144 Tyr358 Ile359 Gly417 and its Leu359 mutant was also investigated in reconstituted systems. Both alleles of P4502C18 were regioselective for 4'-hydroxywarfarin, without any significant stereoselectivity. Both also metabolized warfarin at the 6-position, but to a lesser extent, and metabolism at this site was stereoselective for (R)-warfarin. P4502C8 metabolized warfarin at the 7-position and was stereospecific for (R)-warfarin. It also metabolized warfarin to a lesser extent at the 4'-position, and metabolism at this site was stereoselective for (R)-warfarin. P4502C19 was regioselective for 6 and 8-hydroxywarfarin and was stereoselective for (R)-warfarin. The highly conservative mutation of Ile359 to Leu359 in P4502C9 profoundly altered the regio and stereoselectivity of warfarin metabolism, from regioselective for 7 hydroxywarfarin, with stereospecificity for (S)-warfarin, to regioselective for 4'-hydroxywarfarin, with stereoselectivity for (R)-warfarin, which was confirmed in a reconstituted system using purified recombinant enzymes. In contrast, individual mutations of P4502C9 of Arg144 to Cys, Tyr358 to Cys, and Gly417 to Asp did not markedly affect the regio- or stereoselectivity of warfarin metabolism, although the overall rates of warfarin metabolism were apparently increased by these changes. We conclude that residue 359 is at the substrate binding site of P4502C9, whereas residues 144, 358, and 417, and residue 385 of P4502C18, are not. PMID- 8429827 TI - Binding of the cocaine analog 2 beta-carbomethoxy-3 beta-(4 [125I]iodophenyl)tropane to serotonin and dopamine transporters: different ionic requirements for substrate and 2 beta-carbomethoxy-3 beta-(4 [125I]iodophenyl)tropane binding. AB - The iodinated cocaine analog 2 beta-carbomethoxy-3 beta-(4- [125I]iodophenyl)tropane (beta-[125I]CIT) binds with high affinity to the platelet plasma membrane serotonin transporter, as previously reported for dopamine transporters from rat brain [Eur. J. Pharmacol. 194:133-134 (1991)]. Unlabeled beta-CIT also inhibits serotonin transport by platelet membrane vesicles. In both rat striatal membranes and platelet plasma membranes, beta [125I]CIT binding was found to be pH dependent, with a pKa of 6.4-6.9, and did not require the presence of Cl-. Na+ dramatically stimulated beta-[125I]CIT binding to both serotonin and dopamine transporters, although a small fraction of beta-[125I]CIT binding to the serotonin transporter was observed in the absence of Na+. The substrates serotonin and dopamine competed with beta-[125I]CIT for binding to their respective transporters. However, substrate affinity was enhanced by Cl-, whereas beta-[125I]CIT binding affinity was not. [3H]Imipramine binding to the platelet serotonin transporter and [3H]GBR-12935 binding to the dopamine transporter were not inhibited by decreasing the pH from 8 to 6.5. Likewise, the ability of serotonin to compete with [3H]imipramine binding and that of dopamine to inhibit [3H]GBR-12935 binding were equal at pH 6.5 or 8. Thus, beta-[125I]CIT binding to biogenic amine transporters is distinct from serotonin or dopamine binding by virtue of its inhibition by H+ and its insensitivity to Cl-. PMID- 8429828 TI - Non-neurotoxic amphetamine derivatives release serotonin through serotonin transporters. AB - 3,4-Methylenedioxymethamphetamine (MDMA) and several other amphetamine derivatives cause degeneration of serotonergic nerve terminals. These drugs also release serotonin from nerve terminals both in vivo and in vitro. Two non neurotoxic derivatives of MDMA were tested in membrane vesicle model systems to determine whether they also lacked the ability to release serotonin. 3-Methoxy-4 methylamphetamine (MMA) and 5-methoxy-6-methyl-2-aminoindan (MMAI) both inhibited imipramine binding to serotonin transporters in platelet plasma membrane vesicles and both inhibited Na+ gradient-driven serotonin transport into those vesicles. Significantly, both MMA and MMAI released [3H]serotonin from plasma membrane vesicles, apparently by a process of exchange. The half-maximal concentrations for this effect were comparable to that reported for MDMA. In addition to their effects on plasma membrane transporters, MMA and MMAI both inhibited serotonin transport into chromaffin granule membrane vesicles catalyzed by the vesicular biogenic amine transporter. At higher concentrations, these compounds also caused release of [3H]serotonin from chromaffin granule membrane vesicles and dissipated the transmembrane pH difference (delta pH). Although MMAI effects on the serotonin transporter were similar to those of MDMA, the two compounds had different effects on dopamine transporters. MDMA and methamphetamine inhibited binding of a cocaine analog to the dopamine transporter and released dopamine accumulated by cells expressing dopamine transporters, but similar concentrations of MMAI were inactive. PMID- 8429829 TI - Functional expression of adenosine A2b receptor in Xenopus oocytes. AB - RNA was transcribed in vitro from a cDNA clone (RFL9) that encodes the rat adenosine A2b receptor. Xenopus oocytes that had been injected with this RNA several days earlier responded to adenosine (10 microM to 1 mM) with an inward current (45-750 nA) that peaked rapidly and then declined to a lower level; uninjected oocytes showed no effect of adenosine. The current reversed to outward at -25 mV and was blocked by intracellular injection of 1,2-bis(2 aminophenoxy)ethane-N,N,N',N'- tetraacetic acid. The action of adenosine (100 microM) was mimicked by 5'-N-ethylcarboxamidoadenosine (10 microM), but not by ATP, N6-cyclohexyladenosine (10 or 100 microM), N6-cyclopentyladenosine (10 microM), 1-deaza-2-chlorocyclopentyladenosine (50 microM), or CGS21680 (1 or 10 microM). It was substantially blocked by 8-cyclopentyl-1,3-dipropylxanthine (1 microM) and by 3,7-dimethyl-1-propargylxanthine (10 microM). The results indicate that activation of adenosine A2b receptors increases a calcium-dependent chloride conductance in Xenopus oocytes, presumably by stimulating phospholipase C. PMID- 8429830 TI - Effects of sulphated polyanions on functions of complement factor H. AB - Heparin and two dextran sulphate preparations with a low or high average molecular mass (M(r) 5000 and 5 x 10(5), respectively) enhanced binding of radioactively labelled complement factor H to the complement protein C3b, coupled to Sepharose 4B, maximally 2.5-4-fold within a polyanion concn range of 12.5-400 micrograms/ml. Despite this, heparin or low molecular mass dextran sulphate had no effect on the activity of H as a cofactor of complement factor I, when C3b bound to Sepharose 4B was used as a substrate, and high molecular mass dextran sulphate inhibited. Heparin or low molecular mass dextran sulphate had also no effect on the decay-accelerating activity of factor H on the alternative pathway C3 convertase, C3b,Bb, and high molecular mass dextran sulphate inhibited this activity, too, regardless of whether Sepharose 4B or sheep erythrocytes were used as carriers of C3b,Bb. These results suggest strongly that fluid phase heparin or dextran sulphate do not inhibit activation of the alternative pathway of complement by augmenting functions of H. PMID- 8429831 TI - Monoclonal IgG and IgM autoantibodies obtained after polyclonal activation, show reactivities similar to those of polyclonal natural autoantibodies. AB - In this study, we described the properties of two groups of polyreactive IgG monoclonal antibodies (mAb) derived from splenocytes of a Plasmodium chabaudi infected BALB/c mouse. These IgG mAb reacted with self antigens, but not with the parasite. Depending upon the antigen under study, they showed low (10(-5) M) to high (10(-8) M) affinities. When examined by immunoblotting using mouse organ extracts as the antigen source, each IgG mAb had a different reactivity profile. The IgG2b, but not the IgG2a mAb group, reacted with F(ab')2 fragments of IgG from normal mouse sera, as well as with F(ab')2 fragments of other mAb from the same fusion. Their anti-F(ab')2 activity was inhibited by various antigens, suggesting that the binding sites for these antigens and F(ab')2 fragments were close or overlapping. We also characterized two monoclonal IgM that were strongly and almost exclusively reactive with polyclonal F(ab')2 fragments of normal IgG, and that inhibited the binding of normal polyclonal IgG to self antigens. We previously described such properties for polyclonal IgM from normal mouse sera. The binding of the F(ab')2-reactive IgG mAb to self antigens was also inhibited by normal polyclonal IgM. These results indicate that the IgG and IgM obtained after polyclonal stimulation, exhibit characteristics similar to those of autoantibodies present in normal mouse sera. Furthermore, they confirm our previous studies showing the polyreactivity of polyclonal IgG. Finally, they also show that such IgG mAb exhibit properties similar to those of natural IgM, namely polyreactivity, affinity and connectivity. PMID- 8429832 TI - Effects of brefeldin A on cleavage of invariant chain to p21 and p10 and the appearance of Ii-freed class II MHC dimers. AB - Intracellular cleavage of class II MHC-associated Ii to p21 and p10 and the appearance of Ii-freed alpha, beta dimers were concurrent events lasting from 1 to 6 hr after synthesis of alpha, beta, Ii trimers, possibly related to charging of foreign peptides to the class II MHC antigen-binding site. Sequential immunoprecipitations of pulse-chase radiolabeled cells were made four times with anti-Ii monoclonal antibody to remove Ii and alpha, beta, Ii trimers and then with anti-class II antibody to detect the time-dependent appearance of Ii-freed alpha, beta dimers. The cleavage of Ii to p21 and p10 was revealed in leupeptin treated cells. Cell treatment with Brefeldin A (BFA) was associated with a decrease in Ii-freed alpha, beta dimers, with inhibition of leupeptin-revealed cleavage of Ii to p21 and p10, and with persistence of endoglycosidase H susceptibility of Ii and class II alpha, beta chains. We conclude that in untreated cells, cleavage and release of Ii from class II MHC alpha and beta chains occur after those complexes traverse a BFA-sensitive step in the Golgi apparatus. PMID- 8429833 TI - Cloning and cDNA sequence analysis of nephritogenic monoclonal antibodies derived from an MRL/lpr lupus mouse. AB - Production of IgG3 in MRL/Mp-lpr/lpr (MRL/lpr) lupus mice is one of the major factors to develop glomerulonephritis (GN) in these mice. To examine molecular characteristics of IgG3 responsible for GN in these mice, hybridoma clones producing IgG3 antibodies were prepared from one unmanipulated MRL/lpr mouse. Two clones, 2B11.3 and 7B6.8, were nephritogenic; that is, they caused severe glomerular lesions when injected to normal mice, moreover with a different histopathological manifestation. The 2B11.3 clone generated diffuse cell proliferative lesions, while those induced by the 7B6.8 clone resembled wire loop lesions in human lupus nephritis. The cDNA sequence analysis of 7B6.8 antibody and the other IgG3 antibody, 1G3, non-nephritogenic, revealed that the C regions of the heavy and light kappa chains were completely the same between them. Furthermore, they were identical in deduced amino acid sequences to those from non-autoimmune BALB/c mice, indicating no allelic difference of Igh-8 between these two strains. The V regions of 2B11.3 and 7B6.8 antibodies were composed of different sets of VH, D, JH, Vk and Jk. Although both of the VH belonged to the J558 family, they seemed to use a different VH germline gene. These findings suggest that GN in MRL/lpr mice is generated by the expansion of clonally different B cells producing particular antibodies possibly with a different pathogenetic potency. PMID- 8429834 TI - Characterization and identification of interleukin-1 receptors on bovine fibroblasts. AB - Interleukin-1 (IL-1), a pro-inflammatory cytokine, initiates its many biological effects by first binding to cell-surface receptors. Prior to this study, there were no published reports addressing the nature of the bovine IL-1 receptor. In this study, we characterized and identified cell-surface IL-1 receptors on bovine fibroblasts. Direct binding studies using [125I]-labeled bovine IL-1 beta demonstrated that bovine fibroblasts had approximately 130 high affinity and 2,500 low affinity binding sites (Kd = 4.9 x 10(-11) M and 3.7 x 10(-9) M, respectively). Competitive binding studies using unlabeled recombinant bovine IL 1 beta, IL-2, IFN-alpha, and bovine insulin demonstrated that only unlabeled bovine IL-1 beta competitively blocked fibroblast binding of [125I]-labeled bovine IL-1 beta. Affinity cross-linking of [125I]-labeled IL-1 beta to fibroblasts demonstrated that IL-1 receptors on bovine fibroblasts have an apparent M(r) of 71.5 kD. This report provides the first characterization and identification of IL-1 receptors on bovine fibroblasts. PMID- 8429835 TI - Tyrosine phosphorylation and dephosphorylation in T lymphocyte activation. PMID- 8429836 TI - Comparative effects of ischemia and acute hypoxemia on muscle afferents from tibialis anterior in cats. AB - Comparative effects of ischemia and acute hypoxemia (PaO2 = 24 mm Hg) were studied in anesthetized cats on afferents from the tibialis anterior limb muscle. Metaboreceptors (groups III and IV fibers) and mechanoreceptors were identified by their activation by an intraarterial injection of lactic acid (LA) or high frequency vibrations (HFV) applied to the extremity of the muscle tendon, respectively. Ischemia and hypoxemia exerted opposite influences on the two categories of muscle afferents: they depressed the response of mechanoreceptors to HFV, but markedly enhanced the spontaneous tonic activity of metaboreceptors. The effects of hypoxemia were delayed but slightly greater and lasted longer during the recovery period than those exerted by ischemia. The inhibitory action on mechanoreceptors exerted by a reduced oxygen supply to muscle is interpreted as a result from local acidosis. Indeed, under normoxic conditions, a LA bolus injection during the HFV test also reduced the firing rate of these receptors. PMID- 8429837 TI - Ocular myasthenia gravis: the diagnostic yield of repetitive nerve stimulation and stimulated single fiber EMG of orbicularis oculi muscle and infrared reflection oculography. AB - For the diagnosis of ocular myasthenia gravis (ocular MG), testing of the muscles close to the affected ones may be important. The relative importance of several methods: stimulated single fiber EMG (stimulated SFEMG), repetitive nerve stimulation test (RNS) of orbicularis oculi muscle, and infrared reflection oculography (IROG) was investigated. Thirty-two patients in whom a diagnosis of ocular MG was considered on clinical grounds were admitted to the study. Based on the results of the three neurophysiological tests, the patients could be divided in three groups: a first group with an abnormal stimulated SFEMG, and an abnormal RNS and/or abnormal IROG; a second group with only a slightly abnormal stimulated SFEMG; and a third group with normal tests in all three tests. The clinical diagnosis of ocular MG was made in all 11 patients of the first group; in 86% (6 of 7) of the patients of the second group; and in 7% (1 of 14) of the patients of the third group. This study demonstrates that the orbicularis oculi muscle is a suitable muscle for stimulated SFEMG in patients with ocular MG, and that the results obtained with this technique showed a better relation with the clinical diagnosis than those of the two other techniques. We also demonstrate that there is no additional value in studying the jitter with different stimulation rates in patients with suspected ocular MG. PMID- 8429838 TI - Plasma carnitine insufficiency and effectiveness of L-carnitine therapy in patients with mitochondrial myopathy. AB - Plasma carnitine "insufficiency," (plasma esterified carnitine to free carnitine ratio above 0.25) was found in 21 of 48 (43.8%) patients with mitochondrial myopathy, of whom 4 also showed both total and free carnitine deficiencies in plasma. In addition, plasma levels of SCAC and LCAC were higher in patients with mitochondrial myopathy than in controls (P < 0.001 and P < 0.01, respectively). Patients diagnosed as having plasma carnitine insufficiency or deficiency were treated with L-carnitine (50-200 mg/kg per day in four daily doses). Muscle weakness improved in 19 of 20 patients, failure to thrive in 4 of 8, encephalopathy in 1 of 9, and cardiomyopathy in 8 of 8 patients. Plasma carnitine "insufficiency" provides an additional clue to the diagnosis of mitochondrial myopathy and an indication for L-carnitine therapy. PMID- 8429839 TI - HTLV-1 in acquired adult myopathy. AB - We report a 53-year-old Jamaican man with 20 years of progressing weakness involving proximal limb muscles and neck flexors. Serum CK was 1100 IU/L. EMG demonstrated spontaneous activity, myopathic motor units, and full recruitment patterns in weak muscles. Muscle biopsy revealed marked myofiber degeneration with extensive fibrosis, suggesting a chronic myopathic process. HTLV-1 antibody was present in serum in high titers by ELISA and Western blot. Immunohistochemistry with rabbit polyclonal antisera to HTLV-1 showed rare staining myocytes. PCR demonstrated HTLV-1 DNA in frozen muscle tissue. This myopathy, associated with HTLV-1 infection, has clinical and pathological features similar to a dystrophy. We recommend serological screening for HTLV-1 in cryptogenic adult myopathies. PMID- 8429840 TI - Involvement of peripheral sensory fibers in amyotrophic lateral sclerosis: electrophysiological study of 64 cases. AB - We report electrophysiological findings of conduction along peripheral sensory fibers in 64 patients with amyotrophic lateral sclerosis. Distribution of the values of action potential amplitudes and conduction velocities of peripheral afferent fibers were significantly lower than in normal age-matched controls. Sensory action potential amplitudes (SAPas) were more affected than sensory conduction velocities (SCVs). When single patients were considered, SAPas were slightly but significantly reduced in 22% of the cases (median nerve 17%, ulnar nerve 11%, and sural nerve 22%). A parallel decrease in SCVs and MCVs in 14 patients in whom the study was repeated over a period of time was also found. All these electrophysiological findings are due to progressive neuronopathy of peripheral sensory fibers. A pathogenetic mechanism is proposed. PMID- 8429841 TI - Postmortem examination of relapsing acute Guillain-Barre syndrome. AB - We report a patient who had six episodes of acute Guillain-Barre syndrome during his life. He is the fifth such case with postmortem analysis. PMID- 8429842 TI - Single fiber EMG in a congenital myasthenic syndrome associated with facial malformations. AB - Six patients with a newly described genetic syndrome in Iraqi and Iranian Jews of congenital myasthenia associated with facial malformations were studied with voluntary and stimulation single fiber EMG (SFEMG). Voluntary SFEMG revealed abnormal jitter in all patients in both extensor digitorum communis (EDC) and orbicularis oculi (OOC) muscles, though much smaller in the clinically unaffected EDC. SFEMG study of OOC muscle by axonal stimulation at rates from 1 to 48 Hz showed the most increased jitter at the highest stimulation frequencies in the majority of end-plates, one-third of which showed maximal jitter at intermediate rates. These results may suggest a postsynaptic abnormality as the underlying cause for the neuromuscular transmission defect, and demonstrate the usefulness of SFEMG in the diagnosis of congenital myasthenia. PMID- 8429843 TI - Sustained focal effects of low-dose intramuscular succinylcholine. AB - We studied low-dose intramuscular succinylcholine in 9 subjects as part as an ongoing investigation of its potential to predict responses to botulinum toxin. We measured compound muscle action potentials (CMAPs) from the extensor digitorum brevis (EDB) muscles in each foot before and after intramuscular injections of 2.5 mg of succinylcholine into the EDB. Succinylcholine reduced mean CMAP amplitudes to 42% of baseline; the maximal reduction occurred at 19 +/- 6 (mean +/- standard deviation) minutes. Recovery to 73% of the baseline CMAP amplitude (approximately 50% recovery from block) occurred at 105 +/- 49 minutes after injection. Repetitive (train-of-four) stimulation at 2 Hz produced mild CMAP decrements (5-25%), but only during the recovery phase. Varying the succinylcholine concentrations (10, 20, or 50 mg/mL) while holding the total drug dose constant did not change the rate of onset or the extent of block. No systemic complications occurred. We conclude that: (1) 2.5 mg intramuscular succinylcholine can safely induce selective muscle weakness with a time course that differs from intravenously administered succinylcholine; and (2) further clinical studies comparing intramuscular succinylcholine and botulinum toxin are warranted. PMID- 8429844 TI - Acoustic myography compared to electromyography during isometric fatigue and recovery. AB - This study compared the acoustic (RMS-AMG) to electromyographic (RMS-EMG) signal, median frequency of EMG power spectrum (Fm), and quadriceps torque during isometric fatiguing contraction (FC) and recovery. Seven subjects were tested for strength (MVC) and then, on separate days, maintained 20%, 40%, or 80% MVC to exhaustion followed by MVC testing at regular intervals. Throughout FC, RMS-EMG significantly (P < 0.05) increased and Fm significantly (P < 0.05) decreased during all trials; RMS-AMG significantly (P < 0.05) increased only during the 20% and 40% trials. During recovery, MVC and RMS-EMG recovered most slowly after the 20% trial and most rapidly after the 80% trial; Fm and RMS-AMG recovered by 90 seconds after all trials. RMS-AMG reflects RMS-EMG during low but not high levels of FC. Recovery of strength is most depressed following FC at lower relative levels of torque. We conclude that RMS-AMG behaves differently than RMS-EMG, torque, and Fm during FC and recovery. PMID- 8429845 TI - Factors affecting muscle fiber transformation in cross-reinnervated muscle. AB - Nerves of two fast muscles [peroneus longus (PL) and extensor digitorum longus (EDL)], having different type 2 muscle fiber compositions, were used to cross reinnervate the slow soleus muscle in the rat. Contraction characteristics, histochemical muscle fiber type compositions and myosin heavy chain (MHC) isoform compositions were determined for the reinnervated muscles. Shortening velocity increased in soleus muscles crossreinnervated with EDL nerve [X-SOL(EDL)] but not in muscles cross-reinnervated with PL nerve [X-SOL(PL)]. Type 2A MHC isoform content was increased in X-SOL(EDL) but not in X-SOL(PL), where MHC isoform composition remained similar to normal soleus. The complement of type 1 (slow) muscle fibers was reduced and that of type 2 (fast) fibers increased in both types of X-SOL muscle, but this change was significantly greater in X-SOL(EDL); the majority of the type 2 fibers in X-SOL muscles were of type 2A. Results show that "the type 2 composition" of the reinnervating motoneuron pool is an important factor in determining the transformation of a target slow muscle after cross-reinnervation. PMID- 8429846 TI - Electrophysiological changes in the acute "axonal" form of Guillain-Barre syndrome. AB - The acute "axonal" form of Guillain-Barre syndrome is characterized by rapid progression to severe widespread paralysis and respiratory dependence within 2-5 days of the onset of weakness with very poor and delayed recovery. In 3 cases studied within the first 7 days, the maximum thenar, hypothenar, tibialis anterior, and extensor digitorum brevis "M" potentials were either very reduced in size or absent in response to stimulation at the usual most distal sites of stimulation at the wrist, fibular head, and/or ankle. In the latter instances, advancing the site of stimulation closer to the motor point often evoked an M response. Furthermore, continued distal advance of the site of stimulation evoked progressively larger sized M potentials. Over succeeding days even these very distally evoked M potentials disappeared. Maximum conduction velocities in motor nerve fibers just prior to total loss of excitability were often very reduced. The pattern in these cases is most consistent with progressive loss of excitability and conduction in nerve fibers undergoing axonal degeneration, although coexisting primary demyelination in the terminal segment could not be excluded as the basis for the sometimes very slowed conduction velocities. PMID- 8429847 TI - Use of the surface EMG signal for performance evaluation of back muscles. AB - For well over a decade my associates and I have been developing an objective, noninvasive technique to evaluate the performance of low-back muscles, with emphasis on being able to distinguish between healthy and dysfunctioned backs. Our approach is based on the well-known fact that the EMG signal undergoes a compression in the frequency domain during a sustained muscle contraction. In particular we track the median frequency of EMG signals detected from six muscles in the lower back during an isometric extension of the trunk. The measurements are taken with the Back Analysis System which consists of a postural restraining device, special electrodes for detecting the EMG signals, a muscle fatigue monitor which calculates the median frequencies, and the appropriate software. We have found that the pattern of fatigue exhibited by the six median frequency curves can be used to distinguish individuals who have low-back pain from those who do not with an accuracy of at least 84%. An even more relevant and timely application of our technique is for quantifying the progression of the performance of low-back muscles during a rehabilitation program. Although more work is required to explore the intricacies of the technique, present results provide a convincing indication that it is reliable and that it is ready to be placed into practice. PMID- 8429848 TI - Trichloroethylene does not cause trigeminal neuropathy. PMID- 8429849 TI - Blink reflex measurement of effects of trichloroethylene exposure on the trigeminal nerve. PMID- 8429850 TI - Treatment of multifocal motor neuropathy with a high dosage of intravenous immunoglobulin. PMID- 8429851 TI - Analysis of 462 transplantations from unrelated donors facilitated by the National Marrow Donor Program. AB - BACKGROUND AND METHODS: Allogeneic bone marrow transplantation is curative in a substantial number of patients with hematologic cancers, marrow-failure disorders, immunodeficiency syndromes, and certain metabolic diseases. Unfortunately, only 25 to 30 percent of potential recipients have HLA-identical siblings who can act as donors. In 1986 the National Marrow Donor Program was created in the United States to facilitate the finding and procurement of suitable marrow from unrelated donors for patients lacking related donors. RESULTS: During the first four years of the program, 462 patients with acquired and congenital lymphohematopoietic disorders or metabolic diseases received marrow transplants from unrelated donors. The probability of engraftment by 100 days after transplantation was 94 percent, although 8 percent of patients later had secondary graft failure. The probability of grade II, III, or IV acute graft versus-host disease was 64 percent, and the probability of chronic graft-versus host disease at one year was 55 percent. The rate of disease-free survival at two years among patients with leukemia and good prognostic factors was 40 percent and among patients at higher risk, 19 percent. Twenty-nine percent of the patients with aplastic anemia were alive at two years, and the rate of two-year disease free survival among patients with myelodysplasia was 18 percent. For patients with congenital immunologic or nonimmunologic disorders, the probability of survival was 52 percent. CONCLUSIONS: The National Marrow Donor Program has benefited a substantial number of patients in need of marrow transplants from closely HLA-matched unrelated donors and has facilitated the recruitment of unrelated donors into the donor pool and the access to suitable marrow. PMID- 8429852 TI - Relation between activated smooth-muscle cells in coronary-artery lesions and restenosis after atherectomy. AB - BACKGROUND: Neointimal proliferation leading to restenosis frequently develops after coronary angioplasty. This process is associated with a change in vascular smooth-muscle cells from a contractile (quiescent) phenotype to a synthetic or proliferating (activated) one. We investigated whether the presence of activated smooth-muscle cells in coronary lesions at the time of coronary atherectomy predisposes patients to subsequent restenosis. METHODS: We used in situ hybridization to study the expression of messenger RNA in coronary-atherectomy specimens from 20 patients. Plaque material was hybridized with a probe for the B isoform of human nonmuscle myosin heavy chain, a major nonmuscle myosin isoform in activated, but not quiescent, smooth-muscle cells. Angiographic follow-up data were obtained a mean (+/- SD) of 174 +/- 54 days after atherectomy in 16 of the 20 patients, and the extent of recurrent luminal narrowing was analyzed quantitatively. The presence of restenosis was assessed by exercise thallium scintigraphy in the other four patients. RESULTS: Atherectomy specimens from 10 of the 20 patients showed hybridization with the probe, defined as the clustering of more than 20 silver grains per cell nucleus in more than 10 nuclei in five high-power fields (x250); specimens from the other 10 patients showed no such hybridization. At follow-up, restenosis had developed in 8 of the 10 patients with positive hybridization results, but was absent in 9 of the 10 patients with negative results (P = 0.007). The degree of late loss in luminal diameter was significantly higher in patients with positive hybridization results than in those with negative results (ratio of late loss to immediate gain after atherectomy, 0.76 +/- 0.3 vs. 0.36 +/- 0.3; P < 0.001). CONCLUSIONS: We conclude that the expression of the B isoform of nonmuscle myosin heavy chain is increased in some coronary atherosclerotic plaques and that this increase in expression identifies a group of lesions at high risk for restenosis after atherectomy. PMID- 8429853 TI - Autonomous proliferation of leukemic cells in vitro as a determinant of prognosis in adult acute myeloid leukemia. AB - BACKGROUND AND METHODS: A characteristic of acute myeloid leukemia is the frequent ability of the leukemic cells to sustain their own proliferation in vitro. To determine the clinical importance of this property, we measured the uptake of tritiated thymidine by leukemic cells in serum-free and cytokine-free cultures as a means of determining the rate of spontaneous proliferation in 114 patients with newly diagnosed acute myeloid leukemia. Proliferation was then classified according to three quantitative levels of activity and related to overall survival and to treatment outcome (the response to treatment, the actuarial probability of relapse, and disease-free survival) in 91 patients who were treated with chemotherapy to induce remission. RESULTS: Of the 114 patients, 37 had low, 39 had intermediate, and 38 had high levels of proliferation. The probability of survival at three years was 36 percent among patients with low levels of proliferative activity and 3 percent among those with high levels (P < 0.001). Among the patients treated with chemotherapy, those with low rates of proliferative activity had a 68 percent rate of complete remission and a 49 percent probability of remaining free of relapse, whereas those with high rates of proliferative activity had only a 39 percent rate of complete remission (P = 0.04) and an 11 percent probability of remaining in complete remission (P = 0.009). The probability of disease-free survival at three years among the patients in complete remission after chemotherapy was 49 percent among those with low rates of proliferative activity and 9 percent among those with high rates (P = 0.004). Accordingly, patients with low rates of proliferative activity also had a significantly higher rate of overall survival (44 percent vs. 4 percent; P = 0.002). Patients whose cells had intermediate levels of proliferation in vitro had intermediate rates of survival, relapse, and disease-free survival. CONCLUSIONS: The capacity of leukemic blasts for autonomous proliferation is associated with highly aggressive acute myeloid leukemia. PMID- 8429854 TI - Geographic variation in expenditures for physicians' services in the United States. AB - BACKGROUND: The national volume-performance standard recently implemented by Medicare does not account for geographic variation in expenditures for physicians' services. To study this variation, we examined expenditures for physicians' services in all metropolitan areas in the United States. METHODS: We used Medicare claims data for 1989 to measure rates of service use for beneficiaries living in the 317 U.S. metropolitan statistical areas (MSAs). The variables investigated were rates of admission to the hospital, payments to physicians for inpatient care per admission and per beneficiary, payments to physicians for outpatient care per beneficiary, and overall payments to physicians per beneficiary. Expenditures were measured in terms of allowed charges as adjusted to reflect prevailing charges in each MSA. Rates of use were adjusted for age and sex, with the exception of the variable for payments to physicians for inpatient care per admission, which was adjusted for case mix. RESULTS: Expenditures for the delivery of physicians' services to Medicare beneficiaries varied markedly among MSAs, with those for the areas with the lowest and the highest rates differing at least twofold on each measure. The measures for specific areas varied in parallel: areas with high rates of admission tended to have high levels of payment to physicians for inpatient care per admission, and areas with high payments for inpatient services tended to have high payments for outpatient services. Expenditures were not related to the number of physicians per capita but were lower in MSAs with a high proportion of primary care practitioners. The variation persisted when the 25 largest MSAs were examined; for total payments to physicians per beneficiary, there was a twofold difference between the area with the lowest rate and that with the highest, San Francisco ($872) and Miami ($1,874). The states with the highest overall payments to physicians per beneficiary were Florida, Louisiana, and Michigan. CONCLUSIONS: The marked variation among metropolitan areas in payments to physicians underscores the lack of consensus among physicians about which services are required. Moreover, the practice style in a given community appears to be influenced not by the aggregate supply of physicians but rather by the mixture of primary care physicians and specialists. PMID- 8429855 TI - Seminars in medicine of the Beth Israel Hospital, Boston. Platelet-endothelium interactions. PMID- 8429856 TI - Lucky lady. PMID- 8429857 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 9-1993. A 23-year-old man with fever, dyspnea, extensive bilateral pulmonary infiltrates, and a question of heart failure. PMID- 8429858 TI - Primary care and the affliction of internal medicine. PMID- 8429859 TI - Financing medical education. A universal "Berry Plan" for medical students. PMID- 8429860 TI - The case for internal medicine. PMID- 8429861 TI - Breast augmentation and the risk of subsequent breast cancer. PMID- 8429862 TI - Breast augmentation and the risk of subsequent breast cancer. PMID- 8429863 TI - Breast augmentation and the risk of subsequent breast cancer. PMID- 8429864 TI - Unrealistic expectations in the diagnosis and treatment of ovarian cancer. PMID- 8429865 TI - Unrealistic expectations in the diagnosis and treatment of ovarian cancer. PMID- 8429866 TI - Unrealistic expectations in the diagnosis and treatment of ovarian cancer. PMID- 8429867 TI - Sympathomimetic agents and airway hyperreactivity. PMID- 8429868 TI - Hypercalcemia in pregnancy and lactation associated with parathyroid hormone related protein. PMID- 8429869 TI - Control of epidemics. PMID- 8429870 TI - Prescriptions by mail. PMID- 8429871 TI - Cambridge and Heidelberg compete for new European gene database. PMID- 8429872 TI - Britain plans broad strategy on genome, approves therapy. PMID- 8429873 TI - US panel disagrees on cancer testing method. PMID- 8429874 TI - Diet and cancer. PMID- 8429875 TI - Culture deposits. PMID- 8429876 TI - Sexual selection. Beauty on the brain. PMID- 8429877 TI - Insect development. The long and the short of it. PMID- 8429879 TI - Parkinson's disease. Fetal implants put to the test. PMID- 8429878 TI - Molecular evolution. Genes-in-pieces revisited. PMID- 8429880 TI - Radioactivity of snake venom. PMID- 8429881 TI - Pain pathways and plasticity. PMID- 8429882 TI - Origin of underwater hearing in whales. AB - All described fossil and Recent cetaceans have relatively similar ear bones (malleus, incus and stapes) that strongly diverge from those of land mammals. Here we report that the hearing organ of the oldest whale, Pakicetus, is the only known intermediate between that of land mammals and aquatic cetaceans (whales, dolphins and porpoises). The incus of Pakicetus is intermediate with respect to inflation, crural proportions, and position of the mallear joint. The incus and mandible of Pakicetus indicate that the path of soundwaves to its ear resembled that of land mammals. These fossils suggest that the first whale was amphibious, and corroborate the hypothesis that artiodactyls (for example, pigs, camels and ruminants) are the closest extant relatives of cetaceans. PMID- 8429883 TI - Selection of exaggerated male traits by female aesthetic senses. AB - Darwin suggested that many apparently deleterious secondary sexual characters in males, such as bright colours, elaborate ornaments and conspicuous displays, evolved as a result of female choice. Darwin never tried to explain the crucial agent of selection, that females have preferences for exaggerated male traits. Rather, he took it for granted that females of many species possess a 'sense of the beautiful', akin to the aesthetic sense in humans. The question of why such preferences evolve remains a controversial issue. Here we report that mechanisms concerned with signal recognition possess inevitable biases in response that act as important agents of selection on signal form. The existence of such biases may be sufficient to explain the evolution of exaggerated male secondary sexual traits, and elaborate signals in general. PMID- 8429884 TI - Involvement of an orthologue of the Drosophila pair-rule gene hairy in segment formation of the short germ-band embryo of Tribolium (Coleoptera) AB - The segments in long germ-band insect embryos, like Drosophila, are all determined at syncytial blastoderm stage. This is in contrast to short germ-band embryos which show an early determination of only the anterior head segments, whereas the more posterior thoracic and abdominal segments are sequentially added after formation of a primary germ anlage (reviewed in ref. 1). Segment formation in Drosophila involves the pair-rule genes which define double segmental periodicities and which have been considered to represent a special adaptation to the long germ-band type development hairy belongs to the primary pair-rule genes in Drosophila which are directly regulated by the gap genes, such as Kruppel. We have isolated the orthologues of hairy and Kruppel from the flour beetle Tribolium castaneum which has a short germ type development. We show here that hairy is expressed in several stripes at blastoderm stage and later on in two stripes in the growth zone of the developing embryo. Kruppel expression overlaps hairy stripe three and four expression, very similar to Drosophila. This suggests that the segment patterning mechanism that acts in an open blastoderm in Drosophila works in a similar way in the cellularized Tribolium embryo. PMID- 8429885 TI - Tissue-plasminogen activator is induced as an immediate-early gene during seizure, kindling and long-term potentiation. AB - The requirement of protein and messenger RNA synthesis for long-term memory suggests that neural activity induced by learning initiates a cascade of gene expression. Here we use differential screening to identify five immediate-early genes induced by neuronal activity. One of these is tissue-plasminogen activator (tPA), an extracellular serine protease, which is induced with different spatial patterns in the brain by three activity-dependent events: (1) convulsive seizure increases expression of tPA in the whole brain; (2) stimulation of the perforant path produces an epileptiform after-discharge that ultimately leads to kindling increases the levels of tPA throughout the hippocampus bilaterally; and (3) brief high-frequency stimulation of the perforant path that produces long-term potentiation (LTP) causes an NMDA (N-methyl-D-aspartate) receptor-mediated increase in the levels of tPA mRNA which is restricted to the granule cells of the ipsilateral dentate gyrus. As release of tPA is correlated with morphological differentiation, the increased expression of tPA may play a role in the structural changes that accompany activity-dependent plasticity. PMID- 8429886 TI - DSS4-1 is a dominant suppressor of sec4-8 that encodes a nucleotide exchange protein that aids Sec4p function. AB - The protein Sec4p plays an essential role at the final stage of the yeast secretory pathway and belongs to the ras superfamily of GTP-binding proteins, more specifically to a branch that includes Ypt1p in Saccharomyces cerevisiae and rab proteins in mammalian cells. GTP-binding proteins change conformation depending on whether GTP or GDP is bound and can thus act as a regulatory switch. The protein remains in its inactive, GDP-bound form until exchange of GTP for GDP allows it to stimulate a downstream effector. This interaction is curtailed by GTP hydrolysis. The rates of nucleotide exchange and GTP hydrolysis can be regulated by interaction with accessory proteins. Although GDP dissociation stimulators (GDS) have been identified that act on members of the ras and rho branches of the superfamily, less is known regarding GDSs that act on members of the Sec4/Ypt1/Rab subgroup. A preliminary characterization of a Rab3A GDP dissociation stimulating activity has been presented. We report here the use of suppressor analysis to clone a gene, dss4, encoding a 17K protein that aids Sec4p action in vivo by functioning as a GDP dissociation stimulator. PMID- 8429887 TI - A mammalian guanine-nucleotide-releasing protein enhances function of yeast secretory protein Sec4. AB - Small GTP-binding proteins of the ras superfamily are important for exocytosis from eukaryotic cells. GTP-binding proteins can exist in two different conformations depending on whether they are bound to GDP or GTP, and are thought to function as molecular switches that regulate a variety of cellular processes. The GTP-GDP cycle is controlled by accessory proteins that promote the exchange of bound GDP or the hydrolysis of GTP. The protein Sec4, a member of the Sec4/Ypt1/Rab branch of the Ras superfamily, is involved in a late stage of the secretory pathway in yeast. Here we report the isolation of a mammalian complementary DNA, mss4, encoding a GDP-releasing protein that enhances Sec4 function. The Mss4 protein also stimulates GDP release from Ypt1 and from the mammalian protein Rab3a, but not from Ras2. Mss4 shows sequence similarity to Dss4, a yeast protein with similar biochemical properties. PMID- 8429888 TI - A novel intron site in the triosephosphate isomerase gene from the mosquito Culex tarsalis. AB - The origin and function of introns in eukaryotic genes has provoked considerable debate since their discovery in 1977. Central to this issue are studies on the highly conserved enzyme, triosephosphate isomerase (TPI, EC 5.3.1.1). The 'introns early' argument suggests that introns are as old as the genes themselves and that the apparent correlation of many of the intron sites in plant, animal and fungal TPI genes with the boundaries of modules is evidence of the assembly of ancient proteins by exon shuffling. In contrast, the 'introns late' view holds that ancient genomes contained few if any introns; introns were inserted into pre existing genes during the last billion years. We have found that the TPI gene from the mosquito, Culex tarsalis, contains an intron in a unique position that was predicted by W. Gilbert and the exon shuffling hypothesis. PMID- 8429889 TI - Inhibition of HIV-1 infectivity by zinc-ejecting aromatic C-nitroso compounds. AB - Retroviral nucleocapsid and gag-precursor proteins from all known strains of retroviruses contain one or two copies of an invariant sequence, Cys-X2-Cys-X4 His-X4-Cys, that is populated with zinc in mature particles. Modification of cysteine or histidine residues results in defective packaging of genomic viral RNA and formation of non-infectious particles, making these structures potentially attractive targets for antiviral therapy. We recently reported that aromatic C-nitroso ligands of poly(ADP-ribose) polymerase preferentially destabilize one of the two (Cys-X2-Cys-X28-His-X2-Cys) zinc-fingers with concomitant loss of enzymatic activity, coincidental with selective cytocidal action of the C-nitroso substituted ligands on cancer cells. Based on the occurrence of (3Cys, 1His) zinc-binding sites in both retroviral nucleocapsid and gag proteins and in poly(ADP-ribose) polymerase, we reasoned that the C-nitroso compounds may also have antiretroviral effects. We show here that two such compounds, 3-nitrosobenzamide and 6-nitroso-1,2-benzopyrone, inhibit infection of human immunodeficiency virus HIV-1 in human lymphocytes and also eject zinc from isoalted HIV-1 nucleocapsid zinc fingers and from intact HIV-1 virions. Thus the design of zinc-ejecting agents that target retroviral zinc fingers represents a new approach to the chemotherapy of AIDS. PMID- 8429890 TI - Of mice and men (and cows and cats). PMID- 8429891 TI - Soldiers as experimental animals. PMID- 8429893 TI - Japan begins processing data for genome programme. PMID- 8429892 TI - French suspend child research pending review. PMID- 8429894 TI - UK blood centres could play major role in gene therapy. PMID- 8429896 TI - Universal Darwinism. PMID- 8429895 TI - Selling the future. PMID- 8429897 TI - Universal Darwinism. PMID- 8429898 TI - X-linked immunodeficiency. The fruits of cooperation. PMID- 8429899 TI - Evolutionary biology. The genetics of speciation. PMID- 8429900 TI - Developmental biology. Noggin the dorsalizer. PMID- 8429901 TI - Theories on the brain. PMID- 8429902 TI - Degenerin similarities. PMID- 8429903 TI - Sex surveys and drug users. PMID- 8429904 TI - Protein structure prediction. PMID- 8429905 TI - Haldane's rule has multiple genetic causes. AB - Haldane's rule states that "When in the F1 offspring of two different animal races one sex is absent, rare, or sterile, that sex is the heterozygous [heterogametic or XY] sex". This rule represents one of the few patterns characterizing animal speciation. Traditional explanations of Haldane's rule claim that heterogametic hybrids are unfit because they lack an X chromosome that is 'compatible' with the autosomes of one species. Recent work shows that this explanation is incorrect for hybrid sterility: contrary to prediction, homogametic hybrids carrying both X chromosomes from the same species remain fertile. Until now, similar tests have not been performed for hybrid inviability. Here I show that homogametic hybrids who carry both X chromosomes from the same species are inviable. These results show tht the genetic causes of Haldane's rule differ for hybrid sterility versus inviability. Haldane's rule does not, therefore, have a single genetic basis. PMID- 8429906 TI - Induction of immediate spatiotemporal changes in thalamic networks by peripheral block of ascending cutaneous information. AB - Peripheral sensory deprivation induces reorganization within the somatosensory cortex of adult animals. Although most studies have focused on the somatosensory cortex, changes at subcortical levels (for example the thalamus) could also play a fundamental role in sensory plasticity. To investigate this, we made chronic simultaneous recordings of large numbers of single neurons across the ventral posterior medial thalamus (VPM) in adult rats. This allowed a continuous and quantitative evaluation of the receptive fields of the same sample of single VPM neurons per animal, before and after sensory deprivation. Local anaesthesia in the face induced an immediate and reversible reorganization of a large portion of the VPM map. This differentially affected the short latency (4-6 ms) responses (SLRs) and long latency (15-25 ms) responses (LLRs) of single VPM neurons. The SLRs and LLRs normally define spatiotemporally complex receptive fields in the VPM. Here we report that 73% of single neurons whose original receptive fields included the anaesthetized zone showed immediate unmasking of SLRs in response to stimulation of adjacent cutaneous regions, and/or loss of SLRs with preservation or enhancement of LLRs in response to stimulation of regions just surrounding the anaesthetized zone. This thalamic reorganization demonstrates that peripheral sensory deprivation may induce immediate plastic changes at multiple levels of the somatosensory system. Further, its spatiotemporally complex character suggests a disruption of the normal dynamic equilibrium between multiple ascending and descending influences on the VPM. PMID- 8429907 TI - Cloning and expression of an adenylyl cyclase localized to the corpus striatum. AB - The neurotransmitter dopamine acts through various receptor subtypes that are largely associated with enhancement or inhibition of adenylyl cyclases. These dopamine-sensitive adenylyl cyclases are highly concentrated in the corpus stratum and associated limbic structures of the brain, where their levels exceed by orders of magnitude those in other areas of the brain. Here we use in situ hybridization to show that messenger RNA for three of these adenylyl cyclases is not found in the corpus striatum. We have isolated and expressed a complementary DNA encoding new adenylyl cyclase whose selective concentration in the corpus striatum indicates that it may be responsible for the synaptic actions of dopamine. PMID- 8429908 TI - Nodal is a novel TGF-beta-like gene expressed in the mouse node during gastrulation. AB - During gastrulation, the three germ layers of the embryo are formed and organized along the anterior-posterior body axis. In the mouse, gastrulation involves the delamination of ectodermal cells through the primitive streak and their differentiation into mesoderm. These processes do not occur in embryos homozygous for a retrovirally induced recessive prenatal lethal mutation, the strain 413-d insertional mutation. Instead of giving rise to mesoderm, embryonic ectoderm in 413-d mutants overproliferates and then rapidly degenerates, although extraembryonic lineages remain viable. Here we isolate a candidate for the mutated gene which encodes a new member of the transforming growth factor-beta (TGF-beta) superfamily. Expression is first detected in primitive streak-stage embryos at about the time of mesoderm formation. It then becomes highly localized in the node at the anterior of the primitive streak. This region is analogous to chick Hensen's node and Xenopus dorsal lip (Spemann's organizer), which can induce secondary body axes when grafted into host embryos (reviewed in refs 5 and 6). Our findings suggest that this gene, named nodal, encodes a signalling molecule essential for mesoderm formation and subsequent organization of axial structures in early mouse development. PMID- 8429909 TI - Secreted noggin protein mimics the Spemann organizer in dorsalizing Xenopus mesoderm. AB - A dorsalizing signal acts during gastrulation to change the specification of lateral mesodermal tissues from ventral (blood, mesenchyme) to more dorsal fates (muscle, heart, pronephros). This signal, from Spemann's organizer, cannot be mimicked by the mesoderm inducers activin and fibroblast growth factor. The gene noggin is expressed in the organizer, and could be the dorsalizing signal. Here we show that soluble noggin protein added to ventral marginal zones during gastrulation induces muscle, but that activin does not. Dorsal pattern can be partially rescued in ventralized embryos by injection of a plasmid that expresses noggin during gastrulation. The results suggest that the noggin product may be the dorsalizing signal from the organizer. PMID- 8429910 TI - Cloning and expression of apical membrane water channel of rat kidney collecting tubule. AB - Concentrating urine is mandatory for most mammals to prevent water loss from the body. Concentrated urine is produced in response to vasopressin by the transepithelial recovery of water from the lumen of the kidney collecting tubule through highly water-permeable membranes. In this nephron segment, vasopressin regulates water permeability by endo- and exocytosis of water channels from or to the apical membrane. CHIP28 is a water channel in red blood cells and the kidney proximal tubule, but it is not expressed in the collecting tubule. Here we report the cloning of the complementary DNA for WCH-CD, a water channel of the apical membrane of the kidney collecting tubule. WCH-CD is 42% identical in amino-acid sequence to CHIP28. WCH-CD transcripts are detected only in the collecting tubule of the kidney. Immunohistochemically, WCH-CD is localized to the apical region of the kidney collecting tubule cells. Expression of WCH-CD in Xenopus oocytes markedly increases osmotic water permeability. The functional expression and the limited localization of WCH-CD to the apical region of the kidney collecting tubule suggest that WCH-CD is the vasopressin-regulated water channel. PMID- 8429911 TI - Uncoupling of the molecular 'fence' and paracellular 'gate' functions in epithelial tight junctions. AB - During epithelial morphogenesis, the establishment of tight junctions precedes the development of both the asymmetry in protein and lipid composition between apical and basolateral cell surfaces (the 'fence' function) and the restriction in the transport of ions and nonelectrolytes through the extracellular clefts between cells (the 'gate' function). Molecular models that explain both functions envision strands of particles extending as rings in the cell's perimeter that interact with similar strands located at the apposing cell. This model accounts for the 'fence' function, because the strands prevent diffusion of protein and lipids, and also for the 'gate' function, because the interaction between strands minimizes the width of the extracellular clefts, increasing transepithelial resistance to ions and decreasing non-electrolyte permeability. Here we describe the results of energy depletion, which for the first time separates both functions: it abolishes the gate function, as determined by the dramatic decrease in transepithelial resistance, but it leaves the fence function intact, as determined by the maintenance of lipid polarity. PMID- 8429912 TI - A new factor related to TATA-binding protein has highly restricted expression patterns in Drosophila. AB - The TATA-binding protein TBP is necessary for the transcription of eukaryotic genes. Multi-protein complexes formed by TBP and different TBP-associated factors are involved in the initiation of transcription by polymerases I and II, and probably III as well. During the formation of an active initiation complex, TBP makes specific contacts with other proteins, for example TFIIB and RNA polymerase II (refs 2-4). Here we describe the cloning and characterization of a Drosophila gene product with considerable sequence similarity to TBP and a highly restricted expression pattern in the embryo. This TBP-related factor is a DNA-binding protein but is not likely to be a basal transcription factor. Our results suggest that TBP-related factor is a sequence-specific transcription factor that shares the DNA-binding properties of TBP. PMID- 8429913 TI - How amino-acid insertions are allowed in an alpha-helix of T4 lysozyme. AB - Studies of extant protein sequences indicate that amino-acid insertions and deletions are preferentially located in loop regions, which has traditionally been explained as the result of selection removing deleterious mutations within secondary structural elements from the population. But there is no a priori reason to discount the possibility that insertions within secondary structure could either be tolerated until compensatory mutations arise, or have effects that are propagated away from secondary structure into loops. Earlier studies have indicated that insertions are generally tolerated, although much less well within secondary structure elements than in loop regions. Here we show that amino acid insertions in an alpha-helix of T4 lysozyme can be accepted in two different ways. In some cases the inserted amino acids are accommodated within the helix, leading to the translocation of wild-type residues from the helix to the preceding loop. In other cases the insertion causes a 'looping-out' in the first or last turn of the helix. The individual structural responses seem to be dominated by the maintenance of the interface between the helix and the rest of the protein. PMID- 8429914 TI - Cloning and characterization of a gene that regulates cell adhesion. PMID- 8429915 TI - Multiple-sheathflow capillary array DNA analyser. PMID- 8429916 TI - Antagonists of N-methyl-D-aspartate receptors block seizures induced by putrescine in the deep prepiriform cortex. AB - The role of excitatory amino acid receptors in the genesis of motor and electrocortical seizures, elicited by administration of the polyamine putrescine into the deep prepiriform cortex, has been evaluated in rats. Motor and electrocortical seizures occurred in rats receiving unilateral local injections into the deep prepiriform cortex, of putrescine (10 or 20 nmol). The selective N methyl-D-aspartate receptor antagonist, 2-amino-7-phosphonoheptanoate (AP7), injected previously (15 min) into the deep prepiriform cortex, prevented the development of seizures induced by putrescine, injected at the same site. In addition, dizocilpine (MK-801), a non-selective NMDA antagonist or ifenprodil, a specific inhibitor of the polyamine site at the NMDA receptor, when injected into the deep prepiriform cortex, 15 min prior to putrescine, significantly protected against seizures elicited by this polyamine. A subconvulsant dose of putrescine (5 nmol) potentiated the convulsant effects of NMDA, when injected into the deep prepiriform cortex. These data indicate a potential role of polyamines in the genesis of seizures, elicited from the deep prepiriform cortex. They further suggest that activation of the polyamine site, located at excitatory amino acid NMDA receptors, within the deep prepiriform cortex, may contribute to the genesis of seizure activity in this area. PMID- 8429917 TI - Positron emission tomographic studies of [11C]MDL 72222, a potential 5-HT3 receptor radioligand: distribution, kinetics and binding in the brain of the baboon. AB - The drug MDL 72222, a selective 5-HT3 receptor antagonist, was labelled with 11C and evaluated for distribution kinetics in brain and in vivo binding to 5-HT3 receptors using cold MDL 72222 challenge and positron emission tomography (PET), in three anaesthetized baboons. After tracer doses of [11C]MDL 72222 (i.v. bolus), 11C radioactivity was equally partitioned between plasma and blood cells and readily crossed the blood-brain barrier; it was distributed heterogeneously into 17 different structures of the brain. The kinetic curves for 11C in tissue showed a rapid initial uptake, followed by a slower ascending phase, up to about the twentieth minute and by a plateau, until the end of experiment (90 min). The plateau values indicated marked uptake in brain which, however, varied according to the region considered. In inhibition studies with cold MDL 72222 (1 mg.kg-1) as pretreatment, co-injection or displacement, no clear-cut effects on the kinetics of [11C] MDL 72222 in brain were detected in any region, including those known to be rich in 5-HT3 receptors. These observations suggest that specific binding to 5-HT3 receptors was not detectable in brain in vivo, because of the high lipophilicity (thus a great capacity for non-specific binding) of MDL 72222. These negative findings may also result from both the possible suboptimal affinity of MDL 72222 for 5-HT3 receptors in vivo and the relatively low density of 5-HT3 receptors present only in selected areas of the mammalian brain. This study is a step in the search of selective 5-HT3 receptor radioligands, adequate for in vivo applications. Slow clearance of [11C]MDL 72222 from brain tissue in baboons, should be accounted for in clinical pharmacokinetic investigations for optimal posology considerations. PMID- 8429919 TI - Current bibliographies of neuropeptides prepared by the University of Sheffield Biomedical Information Service. PMID- 8429918 TI - Descending inhibition in the neonate rat spinal cord is mediated by 5 hydroxytryptamine. AB - The inhibitory effects of a stimulus to the thoracic cord on lumbar segmental reflexes were investigated in the superfused cord of the neonate rat. A single stimulus to the latero-ventral cord surface at T11-T12 evoked fast and slow responses in both L4 ventral roots and inhibited rapid segmental reflexes, both ipsi- and contralaterally. The monosynaptic reflex (MSR) was strongly inhibited and the polysynaptic reflex (PSR) and contralateral fast reflex (CON FAST) were inhibited by 30-40%. The inhibition rose to a maximum 2 sec after the conditioning stimulus, plateaued between 2-20 sec and gradually waned to low levels by 100 sec. The slow segmental responses were not inhibited. Inhibition of the MSR was only elicited ipsilaterally and that of PSR was reduced by about 50% on stimulation of the contralateral thoracic cord; inhibition of CON FAST could be evoked from either side of the cord. Inhibition of the MSR from 2-50 sec was greatly reduced by 5-HT2 receptor antagonists. Ketanserin (1 microM) and ritanserin (1 microM) were equally effective but LY 53857 (1 microM) had a weaker blocking action. Only ketanserin reduced inhibition of the PSR. Prazosin (0.1 microM) did not affect inhibition of the MSR but yohimbine (1 microM) blocked it as effectively as ketanserin. This was probably due to 5-HT2 receptor blockade, since 0.1 microM yohimbine had little blocking action and 1 microM idazoxan none, nor did 0.1 microM clonidine mimic inhibition of the MSR. Inhibition of the MSR and PSR was not reduced by 1 microM naloxone, 1 microM strychnine, 1 microM bicuculline nor 10-30 microM APV. Consistent with the release of 5-HT by descending pathways, the 5-HT uptake blocker, citalopram 0.1 microM and the 5-HT releaser, p-chloroamphetamine 1 microM, depressed segmental reflexes, especially the MSR. 5-Hydroxytryptamine did not have the same depressant action on segmental reflexes as stimulation of the thoracic cord; the slow responses were most affected. Both 8-OH-DPAT (1-3 microM) and dipropyl-5-CT (1 microM) preferentially depressed the MSR. Neither spiroxatrine (0.1 microM) nor methysergide (5-10 nM) altered inhibition of the MSR. The concentration of ketanserin required to reduce sub-maximal inhibition by 50% was estimated using 2 concentrations of antagonist. The pIC50, estimated for the blockade by ketanserin of inhibition 20-50 sec after a conditioning stimulus, was 7.3-7.5. It is concluded that inhibition of the MSR and PSR does not involve mediation by glycine, GABAA nor NMDA receptors, nor release of enkephalins nor noradrenaline.(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429920 TI - Nerve growth factor induces a succession of increases in isoprenylated methylated small GTP-binding proteins of PC-12 pheochromocytoma cells. AB - Pheochromocytoma (PC-12) cells exposed to nerve growth factor (NGF) acquire a sympathetic neuron-like phenotype. This NGF-response is blocked by methylation inhibitors and can be mimicked by the farnesylated methylated small GTP-binding protein p21ras. The implicated involvement of prenylation, methylation and a small GTP-binding protein in the NGF-response has been studied by directly measuring 3H-mevalonic acid (MVA)-metabolites incorporated into proteins, protein carboxy [methyl-3H]ester formation and levels of [alpha-32P]GTP-binding proteins in NGF-induced PC-12 cells. We demonstrate that NGF induces a 2-3-fold increase in 21-24 kDa methylated membrane proteins that incorporate 3H-MVA-metabolites, and bind GTP. Levels of [alpha-32P]GTP-binding in these proteins were increased by 2-3-fold. Methylation and membrane association of the small GTP-binding proteins were blocked by lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase, which also enhanced their labeling by 3H-MVA metabolites. Cycloheximide reduced the levels of [methyl-3H] labeled 21-24 kDa proteins and of the overlapping [alpha-32P]GTP binding-proteins. About 70% of the [methyl-3H]-groups found in these proteins were recovered from two dimensional gel blots in nine distinct spots of [alpha-32P]GTP-binding proteins. Taken together these results strongly suggest that in PC-12 cells, NGF induces an increase in the synthesis of prenylated methylated small GTP-binding proteins. The efficacy of lovastatin blockage of protein methylation and enhancement of 3H MVA-metabolites incorporation into GTP-binding proteins was lower in NGF-induced cells than in controls. This suggests that NGF also induces an increase in HMG CoA reductase activity. At the early phase of the NGF response in PC-12 cells (15 min-1 h), the levels of two small GTP-binding proteins (molecular mass of 21-22 kDa and 23-24 kDa) were increased. Thus, at least two proteins, of which one but not the other may be p21ras, appear to be involved in the early response. After a lag period of 24 h with NGF, a second more robust phase of increase in methylated small GTP-binding proteins was apparent. This relatively late response, which was almost completed within 24 h, may reflect involvement of small GTP-binding proteins in neurite-outgrowth and in the functional activity of the differentiated cells. Many small GTP-binding proteins were increased during the second phase, precluding electrophoretic separation of all of them. 3 proteins, however, were well separated (one 23-24 kDa protein and two 21-22 kDa proteins).(ABSTRACT TRUNCATED AT 400 WORDS) PMID- 8429921 TI - Examination of the involvement of neuropeptide Y (NPY) in cerebral autoregulation using the novel NPY antagonist PP56. AB - The contribution of neurotransmitters known to be present in the cervical sympathetic nervous system to cerebral autoregulation was evaluated in the anaesthetised cat using a continuous measurement of cerebral cortical perfusion with laser Doppler flowmetry and an in vitro pial vessel preparation. Autoregulation was tested by venesection and fluid administration to achieve changes in blood pressure from -40% of resting control levels to +80% and flow was monitored. Between -20% and +50% there was no significant alteration in cortical blood flow with perfusion following blood pressure passively outside these ranges. The non-competitive neuropeptide Y antagonist PP56 shifted the level at which the change in flow was passively dependent on blood pressure from +60% to +38%. The pial vessel study demonstrated that PP56 shifted the dose response curve for the vasoconstrictor effect of NPY with a maximal reduction of 22 +/- 6%. These data suggest that the cervical sympathetic nerves with NPY play an active role in cerebral autoregulation. Furthermore in view of the longer time course of action of neuropeptide Y, it is an ideal transmitter candidate to be involved in cerebral autoregulation and any compound that blocks its action must be considered to potentially alter the normal cerebrovascular physiology. PMID- 8429922 TI - Effects of endothelin-1 on postjunctionally-mediated purinergic and adrenergic components of rat vas deferens contractile responses. AB - The effects of endothelin-1 (ET-1) on electrically- or drug-induced contractile responses mediated by purinergic or adrenergic receptors were studied in isolated prostatic portion of rat vas deferens. ET-1 (0.01 nM to 30 nM) concentration dependently increased the contractions evoked by electrical field stimulation (EFS, 0.3 msec, 30 V, 8 Hz, 300 pulses). In the presence of prazosin, ET-1 (3 nM) strongly enhanced the prazosin-resistant responses to EFS, while after desensitization of purinergic receptors induced by alpha,beta-methylene adenosine 5'-triphosphate (mATP) the peptide only tended to enhance the mATP-resistant component of the electrically-evoked contractions. ET-1 failed to change the nonpurinergic nonadrenergic responses to electrical stimulation revealing after simultaneous administration of prazosin and mATP. ET-1 concentration-dependently increased the contractile effects of exogenous ATP (30 microM). The effect of ET 1 (3 nM) was not changed after tetrodotoxin (TTX, 0.3 microM) and guanethidine (10 microM). In the presence of TTX and guanethidine ET-1 potentiated the contractile effects of low (0.01-1 microM) concentrations of noradrenaline (NA) and did not change the contractions induced by NA at concentrations higher than 3 microM. Therefore, ET-1 exerted a potentiating effect on the contractility of the prostatic portion of rat vas deferens via postjunctional mechanisms underlying mainly the purinergic and partly the adrenergic smooth muscle contractile responses. PMID- 8429924 TI - 1993 AAN Membership Directory. PMID- 8429923 TI - Release of immunoreactive neuropeptide Y from brainstem sites in the cat during isometric contractions. AB - Fatiguing isometric contractions of the left hind-limb muscles in cats anesthetized with alpha-chloralose caused mean arterial pressure to increase by 82 +/- 18 mmHg above resting and post-contraction levels and heart rates to increase by 15 +/- 5 beats/min. Contractions were performed by stimulating the tibial nerve via a microprocessor-controlled stimulator. Glass microprobes, coated with antibody specific for neuropeptide Y (NPY) were inserted bilaterally into the periaqueductal grey (P 0.5-1.0, LR 2.0 mm) or into the right ventrolateral medulla (3.0 mm rostral to obex; LR 3.5 mm) prior to, during and following fatiguing contractions to determine whether immunoreactive NPY was released. No release of immunoreactive NPY was detected from the site in the ventrolateral medulla. Immunoreactive NPY was released from the contralateral but not the ipsilateral periaqueductal grey during the isometric contractions, suggesting that NPY-like substances maybe involved with the integration of muscle afferent input into this area of the brainstem. PMID- 8429925 TI - Becoming a better listener. PMID- 8429926 TI - The heart of nursing. PMID- 8429927 TI - Helping the elderly with drug therapy. PMID- 8429928 TI - Teaching your patient to use an incentive spirometer. PMID- 8429929 TI - Suicide survivors. Coping strategies. PMID- 8429930 TI - Assessing a spider bite. PMID- 8429931 TI - Using high-dose morphine to relieve cancer pain. PMID- 8429933 TI - No code: helping the family understand what it means. PMID- 8429932 TI - Why some eye surgery patients are seeing dots. PMID- 8429934 TI - Atherectomy: a different way to unblock coronary arteries. PMID- 8429935 TI - Understanding H.I.V. Why healthy cells self-destruct. PMID- 8429936 TI - Weaning a patient from a ventilator. PMID- 8429937 TI - Do we still need the apothecary system? PMID- 8429938 TI - Trying to rescue Frank. PMID- 8429939 TI - Caring for E.D. patients: 13 ways to protect your practice. PMID- 8429940 TI - Judy was an expert on diabetes ... and noncompliance. PMID- 8429942 TI - Myths & facts ... about charting. PMID- 8429941 TI - Evaluating serum calcium levels. PMID- 8429943 TI - Self-test. Calculating drug dosages. PMID- 8429944 TI - Managing a perforated viscus. PMID- 8429945 TI - Health challenges for the 1990s. PMID- 8429946 TI - Trends in AIDS mortality among residents of Puerto Rico and among Puerto Rican immigrants and other Hispanic residents of New York City, 1981-1989. AB - This study contrasts the acquired immunodeficiency syndrome mortality experience of residents of Puerto Rico with that of New York City residents identified as either Puerto Rico-born and non-Puerto Rico-born Hispanics. Portions of the mortality data examined in this investigation update and extend the data previously published describing selected groups in New York City through the end of 1987 but which did not consider residents of Puerto Rico. The nine-year cumulative, age-adjusted acquired immunodeficiency syndrome mortality rate for males was found to be 5 x higher among Puerto Rico-born New York City residents compared with residents of Puerto Rico (702/100,000 v 141/100,000) and 1 1/2 x greater than that of other male Hispanic New York City residents (447/100,000). In New York City, Puerto Rico-born females had higher age-adjusted mortality rates (121/100,000) than female residents of Puerto Rico (25/100,000) and other female Hispanic residents of New York City (70/100,000). Within five of the six age categories considered, acquired immunodeficiency syndrome mortality rates for adult males and females are higher for Puerto-Rico-born, New York City residents. Limitations of acquired immunodeficiency syndrome incidence data, as they pertain to persons of Puerto Rican ancestry, are discussed. PMID- 8429947 TI - Anti-convulsant drugs, smoking, and body weights in psychiatric in-patients. AB - Some anti-convulsant drugs have a calming effect that may potentially be used to reduce cigarette smoking. A cross-sectional study, including all 100 psychiatric in-service patients receiving various anti-convulsant drugs and their age- and sex-matched controls, was done. The intensity of their daily cigarette smoking and their body weights were recorded. Of the patients, 25% were non-smokers. Of the 22 patients on phenytoin, eight were non-smokers as opposed to three in the control group. Two thirds of the phenytoin group patients were either non smokers, or were nominal smokers. As opposed to this, of the control patients and those on other anti-convulsants, two thirds were either moderate or heavy smokers. The phenytoin group of patients weighed less than the controls (149 v 163 lbs). Valproic acid therapy was associated with a significantly higher body weight and with more smoking. Clonazepam's effect was similar to valproic acid, but carbamazepine did not show any relationship either with smoking or with body weights. Therapy with phenytoin is associated with a lower prevalence and a lower intensity of cigarette smoking, together with lower body weights in psychiatric in-patient population. PMID- 8429948 TI - State experience with Medicare hospital mortality: how does New York State compare? AB - 1987 Medicare hospital mortality statistics for 255 New York hospitals (including 57 hospitals in New York City) were compared with 4,617 hospitals located in other states. An analysis of covariance examined how overall Medicare hospital mortality rates differed across states; these rates were adjusted for expected mortality, hospital bed size, and major teaching status. This study tested for the hypothesis that New York State had an average 30-day post-admission Medicare mortality rate significantly different from the mean hospital mortality rate for all states. The results indicated that New York State was -0.43 percentage points below the average risk-adjusted mortality rate of the other states (p < 0.0001). Although it remains speculative to what extent differences in adjusted Medicare hospital mortality are a function of quality of care, these results indicate that New York's historically high level of hospital regulation has not resulted in inferior patient outcomes. PMID- 8429949 TI - Sudden death after acute myocardial infarction. PMID- 8429950 TI - Ethical and social issues in organ procurement for transplantation. PMID- 8429952 TI - Cost containment and academic medicine. PMID- 8429951 TI - The story of Lassa fever. Part IV: The politics of research. PMID- 8429953 TI - Scientific failure in an age of optimism: public reaction to Robert Koch's tuberculin cure. PMID- 8429954 TI - Molecules and magnets. PMID- 8429955 TI - Psychic advisor. PMID- 8429956 TI - Mediastinal teratoma presenting as empyema. PMID- 8429957 TI - Missed diagnosis of acute arthritic gout in kidney transplant recipients. PMID- 8429958 TI - Supraventricular tachycardia induced by swallowing. PMID- 8429960 TI - Transitional cell carcinoma of the renal pelvis in an incompletely duplicated collecting system. PMID- 8429959 TI - Morgagni hernia mimicking a lipomatous tumor. PMID- 8429961 TI - Distress over attempts to discontinue the New York State Journal of Medicine. PMID- 8429962 TI - The need to continue publishing the New York State Journal of Medicine. PMID- 8429964 TI - 1993 House of Delegates members. PMID- 8429963 TI - The status of the New York State Journal of Medicine: where are we? PMID- 8429965 TI - Medicare reimburses outpatient diabetes education. PMID- 8429966 TI - Support for independent transcription. PMID- 8429967 TI - Legal update for nurses, Part II: Assigning, delegating and staffing. PMID- 8429968 TI - Mothers, infants and children: what's happening now? PMID- 8429969 TI - Second RBRVS fee year arrives. PMID- 8429971 TI - The Executive Information System's basic eight. PMID- 8429970 TI - Abortion: a tangle of rights. PMID- 8429972 TI - Strategic planning: a practical approach. AB - Strategic planning identifies a department's environmental factors while establishing its purpose, mission, philosophy and goals and developing a long range plan and objectives. This hospital's practical approach mirrors the nursing process related to assessment, diagnosis and planning, implementation and evaluation. PMID- 8429973 TI - Assessing organizational culture: a planning strategy. AB - Three semi-urban hospitals in Oregon were surveyed to assess the strength of various organization and work unit ideologies and the extent of cultural homogeneity. Occupational cultures were consonant with the larger organization in two hospitals, but the third showed dissonance. This affected management strategies for conflict resolution, staff development and organizational change. PMID- 8429974 TI - Integrating research in a strategic plan. AB - It is important that the philosophy and strategic plan for a Department of Nursing support research programs which will guide current clinical and administrative decisions as well as long-term policy initiatives. Research also contributes to the maturing of the science of nursing through generating and testing knowledge. PMID- 8429975 TI - Managing HIV-positive and AIDS risks: educational and psychosocial resource assessment. AB - A needs assessment survey reveals many staff nurses incorrectly estimate the HIV contagion risk, inadequately follow universal precaution mandates, and are often unaware of available information and counseling resources. Nurses want current, reliable HIV-AIDS related information and counseling from credible, accessible, and readily available sources. Colleagues, families, and friends' attitudes decidedly influence nurses' capacity to cope emotionally. PMID- 8429976 TI - The assessment center: evaluating managerial potential. AB - This assessment center helps unit directors critique their own managerial strengths and weaknesses and formulate effective self-improvement plans and growth strategies. Structured activities also provide a framework for administrators to identify managerial talent and to develop potential nurse managers effectively. PMID- 8429977 TI - Supervising the co-dependent nurse. AB - Co-dependent behaviors influence not only the effectiveness of the individual nurse involved, but the entire work environment as well. Recognizing such patterns and using proven techniques to manage these behaviors can help nurse managers encourage nurses to move towards autonomous, healthy functioning. PMID- 8429979 TI - Thriving on chaos. PMID- 8429978 TI - The nurse advocate and care for the caregivers. AB - Dramatic improvement in morale, increased retention and an enhanced atmosphere of optimism resulted from introduction of a Nurse Advocate role in an acute care hospital. Within the context of a collaborative governance model, the Nurse Advocate was able to empower individuals and groups to immediate successes. Use of registry nurses was reduced by 99 percent! PMID- 8429980 TI - Service excellence: unit-based service objectives in CQI. AB - The healthcare industry is joining business through redefining the patient as "customer". High-quality customer service requires a high measure of personal attention plus dependability, promptness and employee competence. With a motto of "People First," this organization pledged itself to "consistently meet or exceed our customers' personal and practical needs in a courteous, accurate and timely manner." " PMID- 8429981 TI - Combining patient classification and nursing documentation. PMID- 8429982 TI - A physician's view on nurse-doctor progress notes. PMID- 8429983 TI - Adjusting primary nursing to 12-hour shift. PMID- 8429984 TI - Research in a clinical setting. PMID- 8429985 TI - LPNs as medication nurses. PMID- 8429986 TI - 201Tl-labelled TlCl dosimetry revisited. AB - A review of the literature pertaining to the human dosimetry of 201Tl-labelled thallous chloride provides a spectrum of organ absorbed doses associated with this radiopharmaceutical. A more recent article details human quantitative studies up to 216 h postadministration. This multiorgan radiopharmacological study, however, reports the kidney dosimetry to be significantly lower than previous values; 0.0647 mGy MBq1 (0.238 rad mCi-1) compared to 0.326 mGy MBq-1 (1.2 rad mCi-1). Because of the latter discrepancy the dosimetry was recalculated using the reported pharmacodynamic data. This recalculation resulted in a kidney radiation dose comparable to previously published reports of 0.514 mGy MBq-1 (1.89 rad mCi-1). Also, revised absorbed doses to the various segments of the gastrointestinal tract (GIT) were significantly higher. PMID- 8429987 TI - Secretion of radioactivity in breast milk following administration of 99Tcm-MAG3. AB - Measurements were made of the fraction of injected radioactivity which was secreted in breast milk following 99Tcm-MAG3 dynamic renal scans of two nursing mothers. The activity concentration in the milk expressed at different times after injection was found to decrease monoexponentially, with effective half lives of 3.2 and 4.0 h. Two models were applied to each set of data and predicted an overall mean infant effective dose equivalent per unit administered activity of 2.9 muSv MBq-1 and maximum of 4.5 muSv MBq-1. The results demonstrated that it was not essential to interrupt breast feeding following administration of 100 MBq 99Tcm-MAG3 for a dynamic renal scan. If a renal perfusion scan is performed using greater activities of 99Tcm-MAG3, the mother should interrupt feeding and not resume until measurements on her expressed milk samples indicate that the effective dose equivalent to the infant will be less than 1 mSv. However, it was noted that a considerably lower radiation dose to the infant could be achieved by using 99Tcm-DTPA. Therefore, it was concluded that whenever clinically appropriate 99Tcm-DTPA should be used for both types of renal investigations of breast-feeding mothers. PMID- 8429988 TI - Intraoperative bone and bone marrow sampling: a simple method for accurate measurement of uptake of radiopharmaceuticals in bone and bone marrow. AB - Accurate estimation of bone marrow uptake of radiopharmaceuticals is of crucial importance for accurate whole body dosimetry. In this study, a method for obtaining normal bone marrow and bone during routine surgery without inconvenience to volunteers is suggested and compared to an indirect method. In five volunteers (group 1), 4 MBq 111In-labelled human polyclonal IgG (111In-IgG) was administered 48 h before placement of a total hip prosthesis. After resection of the femoral head and neck, bone marrow was aspirated from the medullary space with a biopsy needle. In five patients, suspected of having infectious disease (group 2), bone marrow uptake was calculated according to a well-accepted method using regions of interest over the lumbar spine, 48 h after injection of 75 MBq 111In-IgG. Bone marrow uptake in group 1 (4.5 +/- 1.3% D kg-1) was significantly lower than that in group 2 (8.5 +/- 2.1% D kg-1) (P < 0.01). Blood and plasma activity did not differ significantly for both groups. This method provides a system for directly and accurately measuring uptake and retention in normal bone marrow and bone of all radiopharmaceuticals at various time points. It is a safe and simple procedure without any discomfort to the patient. Since small amounts of activity are sufficient, the radiation dose to the patient is low. PMID- 8429989 TI - Cerebral perfusion deficits in divers with neurological decompression illness. AB - Cerebral perfusion deficits detected by injection of 99Tcm hexamethylpropyleneamine oxime (HMPAO) and single photon emission tomography is said to correlate well with clinical findings in divers with neurological decompression illness. We studied 12 divers. Six had residual cerebral signs (group 1) and six had no residual cerebral symptoms or signs (group 2). Perfusion deficits were as common in group 2 as in group 1. The site of the deficit did not correlate well with either the neurological findings at presentation or the residual clinical signs after treatment. The data suggest that claims that HMPAO scanning correlates with clinical findings and can be used for patients management were incorrect. PMID- 8429990 TI - The feasibility of 225Ac as a source of alpha-particles in radioimmunotherapy. AB - This paper proposes the utilization of 225Ac for the alpha-radioimmunotherapy of cancer. The isotope decays with a radioactive half-life of 10 days into a cascade of short-lived alpha- and beta-emitting isotopes. In addition, when indicated by the pharmacokinetic requirements of particular clinical applications, 213Bi, with a radioactive half-life of 47 min, can be chosen as an alternative source of alpha-particles in radioimmunotherapy. This isotope is the last alpha emitter in the 225Ac decay-cascade and can be extracted from a 225Ac source at the bedside of the patient. 225Ac can quasi ad infinitum be obtained from one of its precursors, 229Th, which can be made available by various means. The indications for the use of alpha-particles as an alternative to more traditional classes of radiation are derived from the particle-kinetic characteristics and the radioactive half-life of their source isotope, as well as from the properties of the target-selective carrier moiety for the source isotope. It may be expected that useful applications, complementary to and/or in conjunction with other means of therapy will be identified. PMID- 8429991 TI - Out-of-hours scintigraphy: a survey of current practice. AB - A questionnaire was sent to 221 nuclear medicine departments in the UK asking about their staffing and work patterns. In particular, we wanted to know how many of them offered an on-call service. In those cases where departments wished to offer this service but did not do so, they were asked what was stopping them. Replies were received from 150 departments, a response rate of 68%. Of these, 43 (29%) offered an on-call service, although only 21 (14%) performed ten or more out-of-hours scans in an average year. The examinations most commonly offered were lung scans, localization of gastrointestinal bleeding and renography. In those centres wishing to offer an on-call service but which were unable to do so, limitation of resources was the reason most frequently advanced. Lack of clinical demand was the factor most often quoted by those not wishing to extend their service. The issue of emergency scintigraphy is discussed in the light of these results. PMID- 8429992 TI - Changes in parathyroid hormone during haemodialysis sessions with two different dialysis membranes: specific adsorption of intact parathyroid hormone. AB - In patients dialysed with two different membranes (cuprophan and high-flux polysulphone) and using three radioimmunoassays recognizing either intact molecule (iPTH), C terminal (cPTH) or median fragments (mPTH), we compared parathyroid hormone levels at the beginning of and during dialysis. At the beginning of dialysis, cPTH and mPTH levels were always increased but iPTH levels were sometimes within the normal range; during dialysis only iPTH distinctly decreased whichever membrane was used. The behaviour of iPTH cannot be explained by crossing through the membrane because its molecular weight is too high; it cannot be explained by the variation of calcium and phosphate parameters during the session because no correlation exists between the variations in plasma levels of iPTH and the variations in phosphorous and calcium levels. In vitro experimentation with radiolabelled intact iPTH was performed to confirm the specific behaviour of iPTH: these experimental data agree with our in vivo results and suggest an adhesion to the dialysis membrane. Our study shows a specific singular property of iPTH during dialysis and demonstrates the necessity of considering the time of sampling during dialysis and the moiety of parathyroid hormone being measured in patients undergoing dialysis in order to assess osteodystrophy. PMID- 8429993 TI - 123I-SCH 23982 is not suitable for dopamine D1 receptor imaging in vivo in the human brain. AB - The tracer 123I-SCH 23982 was tested with regard to its ability to image dopamine D1 receptor in the human brain in vivo with single photon emission computed tomography (SPECT). The tracer did not reach equilibrium with regard to its binding to dopamine D1 receptors, presumably owing to fast metabolism to hydrophilic products and deiodination. It is concluded that 123I-SCH 23982 is not suitable for dopamine D1 receptor imaging with SPECT in the human brain. PMID- 8429994 TI - Immunoscintigraphy in malignant melanoma: a five-year clinical experience. AB - Immunoscintigraphy (IS) with 99Tcm-labelled anti-melanoma monoclonal antibody F(ab')2 fragments was performed in 135 melanoma patients, 64 males and 71 females, aged 19-82 years (mean 52.3 years) between December 1987 and December 1991. The first group of IS was performed in 50 patients before surgery to assess optimal management: seven true positive and one true negative were obtained in ocular and visceral melanomas, while in cutaneous MM sensitivity, specificity and accuracy in assessing lymph node involvement were, respectively, 61.5, 93.3 and 83.7%. The second group of 128 IS is relative to 85 patients in follow-up: excluding 13 cases with known metastatic disease and 12 inconclusive tests, sensitivity, specificity and accuracy were, respectively, 83.3, 98.8 and 96.1%. Immunoscintigraphy is free of side effects even after repeated administrations and is a useful adjunct to standard diagnostic techniques as a basis for treatment decisions. PMID- 8429995 TI - Technetium-labelled neutrophils in rheumatoid arthritis. PMID- 8429996 TI - Towards standardization in gamma camera renography. PMID- 8429998 TI - Does radioiodine cause the ophthalmopathy of Graves' disease? PMID- 8429997 TI - Towards standardization in gamma camera renography. PMID- 8429999 TI - Exercise 201Tl scintigraphy: evaluation of the additional diagnostic value. AB - The additional diagnostic yield of exercise 201Tl scintigraphy using both visual and quantitative analysis was determined in 221 patients with known or suspected coronary artery disease (CAD). The coronary arteriogram was adopted as the gold standard. After pretest clinical and exercise electrocardiographic data were taken into consideration, scintigraphy added diagnostic accuracy both in the diagnosis of CAD and of multivessel disease. The diagnostic yield of the scintigraphy in terms of sensitivity and specificity was, however, not significant. In 79% (121/153) of the patients, the diagnosis of the presence, or exclusion, of CAD was highly probable (P > 0.80 or P < 0.20) when considering clinical and exercise data. The diagnosis was, however, not significantly improved by the scintigraphic result. Twenty-seven per cent (20/73) returned a negative scintigraphic results with a high (P > 0.80) prescintigraphic probability for CAD and a positive arteriogram. It was concluded that 201Tl scintigraphy has additional diagnostic value after clinical and exercise parameters were taken into consideration in the diagnosis of coronary artery and multiple vessel disease. It is not recommended to refer patients with either a low or high probability of CAD for screening and diagnosis as in a high proportion of patients the diagnosis of CAD could have been made using clinical or exercise data alone. PMID- 8430000 TI - 201Tl as a tumour-localizing agent: current status and future considerations. PMID- 8430001 TI - Nursing's agenda for health care reform: what do Americans believe about nursing? PMID- 8430002 TI - Questions and answers about AIDS. PMID- 8430003 TI - Intraocular pressure in a selected sample of myopic and nonmyopic Chinese children. AB - The intraocular pressure (IOP) measurements for 30 myopic and 30 nonmyopic children between 6 and 9 years of age were extracted from eye examination records. The mean IOP of 30 myopic children was 13.69 mm Hg (SD 3.4) and that of 30 nonmyopic children was 11.55 mm Hg (SD 2.06). The difference between the two is significant. These results suggest that IOP may play a role in the determination of refractive error. PMID- 8430004 TI - Effects of early diabetic retinopathy on rod system sensitivity. AB - Previous studies have shown that S-cone pathway sensitivity is selectively decreased in the early stages of diabetic retinopathy. In the present study, rod system sensitivity was evaluated in a group of diabetic patients using psychophysical techniques. The course of dark adaptation was first determined, then absolute thresholds were measured in the horizontal and vertical meridians. For all patients, although the recovery of the initial portions of rod dark adaptation were normal, absolute thresholds were increased in both the horizontal and vertical meridians. The findings provide evidence that patients with early diabetic retinopathy show a generalized dysfunction of the rod system. PMID- 8430005 TI - Effect of color on contrast sensitivity with two different accommodative stimuli. AB - We studied the influence of color and accommodation on the contrast sensitivity function (CSF). At the same time, we measured the effect of axial chromatic aberration (ACA) on the CSF. The CSF's of two observers were determined using red, green, blue, and white light, at 5- and 0.5-m viewing distances. At 5 m the CSF's were measured with natural vision and also with lenses which compensated the ACA. Results show that the effect of ACA on the CSF is to reduce the sensitivity to blue with respect to the red. The difference in sensitivity between these two colors is between 50 and 150% and varies with the frequency and the subject. When the ACA is compensated the influence of the color on the CSF is very small, in our experiment, and this result is discussed in relation to the compensating effects of chromatic adaptation of the color mechanisms. In near vision, the CSF's were measured with natural vision and also with the lenses required to give the same accommodative stimulus for all colors. We discuss the effect of accommodation on the CSF. For both far and near the results are compared with the CSF with white light. PMID- 8430006 TI - Review of the tear break-up time and a closer look at the tear break-up time of Hong Kong Chinese. AB - The technique of assessing tear stability using the fluorescein instillation technique has been widely used in the clinical diagnosis of "dry eye" and in the screening of potential contact lens wearers. Various inconsistencies in the application of the tear break-up time (TBUT) techniques and the results obtained were noted in the literature and this review summarizes the differences and the conclusions of various investigators. The reliability of this technique using two different methods of observation was examined and the mean TBUT of Hong Kong Chinese subjects (HK-Chinese) was determined. We conclude that the TBUT can be measured reliably using either the scanning or full beam method in most patients. The mean TBUT of HK-Chinese is approximately 8 s and about 70% of HK-Chinese have TBUT's less than 10 s. PMID- 8430007 TI - Discrepancy in the evaluation of visual impairment of elderly low-vision patients by general eye care practitioners and by low-vision practitioners. AB - Levels of impairment and disability determine eligibility for different welfare programs for visually handicapped persons. General eye care practitioners have to determine the level of impairment for administrative purposes, whereas the low vision practitioner measures the level of impairment in order to plan the rehabilitation program. We compared the severity of visual impairment reported by the referring practitioners and the one reported by the low-vision specialists for the population of elderly low-vision patients receiving care from one of the largest rehabilitation centers for the visually handicapped in Quebec. Visual acuity ratings reported by general practitioners are lower than those reported by low-vision practitioners. The presence of visual handicap is generally well identified but the severity of visual impairment according to the World Health Organization (WHO)'s categories is not properly gauged by the general eye care practitioners as expressed by the sensitivity and the positive predictive value of the classification of the general practitioners in respect to the classification of the low-vision optometrists. The need to use existing standardized methods to measure visual acuity (VA) is emphasized. It may be hypothesized that this overestimation of the severity of the visual condition may have a negative impact on the process of rehabilitation of the elderly. PMID- 8430008 TI - Repeated visual acuity measurement: establishing the patient's own criterion for change. AB - We measured visual acuity in 10 young subjects, 10 times each over a period of approximately 3 weeks, using Bailey-Lovie charts. We used a consistent end-point criterion and scored each letter read on the chart. We derived the mean and standard deviation of visual acuity measures for each subject, and for the group. The standard deviation for the group was about 3 times that of the individuals in the group. We calculated the criterion for reduction of visual acuity for the group, as group mean plus 1.96 group standard deviations; use of this criterion would consistently fail to detect patients with clinically significant reductions in visual acuity. We recommend that visual acuity be measured to threshold for every patient. Measuring visual acuity between three and five times provides an estimate of the patient's variability and allows a criterion for reduction of visual acuity to be established for the individual patient. Use of this criterion will enhance the sensitivity of visual acuity measurement as a diagnostic tool. PMID- 8430009 TI - Visual requirements for reading. AB - We have applied research on the visual psychophysics of reading to low vision assessment. Research on different aspects of the reading process found that reading rate rather than reading comprehension is more sensitive to variations in a subject's visual functioning or the stimulus properties of print. The research identified four different visual factors that significantly affect reading rate: (1) acuity reserve [print size relative to acuity threshold], (2) contrast reserve [print contrast relative to contrast threshold], (3) field of view [number of letters visible], and (4) in cases of maculopathy, central scotoma size. Our research indicates that fluent reading rates can be attained with a restricted field of view, as little as four characters. However, attainment of fluent reading levels requires that print size and contrast should be several times threshold and the diameter of a central scotoma should be less than 22 degrees. Although important clinical studies are lacking, we derived specific visual requirements for different reading rates from published experimental research to provide a starting point and to illustrate how visual requirements could be derived, even with poor correlations. Research has made significant progress toward the development of a comprehensive low vision assessment that will allow the practitioner to identify visual impediments to reading, other than reduced visual acuity. Having more fully characterized a visual impairment, the practitioner may tailor devices or interventions to the individual's needs and capabilities. PMID- 8430010 TI - Comparison of the transmittance and coloration requirements of the four national sunglass standards. AB - There are four national standards on nonprescription sunglasses and two international draft standards at different levels of development and activity. The variation in test specifications and pass/fail criteria among the national standards makes comparisons difficult and agreement on international standards slow. Arguments about the relative stringency of standards may be, as a consequence, ill-informed and self-defeating. In this study I compared the requirements of the four national sunglass standards and then the compliance of 253 sunglass and fashion spectacle lenses with the standards in the erythemal ultraviolet (UV), near UV, luminous transmittance, and coloration. The methods and criteria of the four standards vary quite substantially but the lenses failing to comply are frequently the same. The differences and similarities among the standards are clearly demonstrated. In the erythemal UV, the standards form two pairs in their stringency (Australian and British Standards as against German and U.S. Standards). In the near UV, the British Standard stands out as being more stringent. In the luminous transmittance region there is a basic agreement. In coloration requirements, even though there is extreme variation of methods, these too provide very similar results. The U.S. requirements on signal transmittance do not serve a useful function. Some comments on the differences in labeling requirements are made. PMID- 8430011 TI - Pigmented paravenous retinochoroidal atrophy. AB - Pigmented paravenous retinochoroidal atrophy is a rare disorder that is not well understood or classified. We examined a patient who had chorioretinal atrophy with pigment clusters located in the paravenous areas without macular involvement. Other conditions which may also present retinochoroidal atrophy and/or pigmentary degeneration are discussed. The electroretinography results were subnormal but the evoked potentials were not totally extinguished. Fluorescein angiography and retinography confirmed the relative and absolute scotomas corresponding to the atrophic paravenous areas. PMID- 8430012 TI - Statistics notebook: entry IV.B: (1) one-sample t-test and (2) matched-pairs t test. PMID- 8430013 TI - Brake light on automobiles. PMID- 8430014 TI - Management of contact lens patients with pingueculae or pterygia. AB - Diplomates in the Contact Lens Section of the American Academy of Optometry were surveyed to determine how they managed contact lens patients with pingueculae or pterygia. Responses indicated that many patients with these conditions can be successful contact lens wearers. Specific management techniques used by these practitioners are reported. PMID- 8430015 TI - [The role of thoracic radiography in the intensive care unit of a department of internal medicine]. AB - Authors review the main principles of chest X-ray examination methodology and analyse their one-year material. During this time 225 chest X-ray films were taken by means of a mobile X-ray unit. The majority of patients were treated for cardiac disorders (e.g. AMI). Authors analyse the distribution of patients according to their illnesses, the correlation between the clinical and the radiological diagnoses and the quality of the X-ray films. Authors emphasize the importance of the correct collaboration between the staff of the intensive care unit and the radiological department. PMID- 8430016 TI - [Lasers in otologic surgery]. AB - The author makes known the possible advantages of the laser-application in the ear surgery. He informs about the main tissue effects of the surgical lasers used in practice. On the bases of the bibliography and his own experiences the author offers a survey of the indications of laser-oto-surgery. PMID- 8430017 TI - [Treatment of carcinoid tumor with a long-acting somatostatin analog]. AB - A case of carcinoid tumour of the small intestine has been reported, which caused hormone producing multiple hepatic metastases. The 45 year old man had flush syndrome several times a day, that was caused by a carcinoid tumour originated from the jejunum. After the operation of the primary tumour he was treated by Sandostatin (Sandoz, Basel), which significantly reduced the symptoms and slowed down the progression. The neural elements containing substance P, neuropeptide Y, vasoactive intestinal polypeptide, serotonin, dopamine-beta-hydroxylase and somatostatin, which are thought to cause the symptoms, were investigated in the small intestine and the liver metastasis by an immunocytochemical method. It seems to be an interesting observation, that a large number of neuropeptide Y immunoreactive nerve fibers and some substance P and vasoactive intestinal polypeptide immunoreactive nerve processes and cell bodies were observed however, no trace of somatostatin and serotonin immunoreactive nerve elements could be detected in the hepatic metastasis. That can be a novel addition to the possible aetiology of the carcinoid syndrome and to the way how the somatostatin treatment acts. PMID- 8430018 TI - [The medical and medico-historical value of the periodical Magyar Szemle]. PMID- 8430019 TI - [Lasting value of a 150-year-old principle: the Doppler effect]. PMID- 8430020 TI - [Endoscopic and surgical removal of foreign bodies from the upper gastrointestinal tract]. AB - Results of management of 67 patients who swallowed foreign bodies are discussed. Types of foreign bodies, the importance of previous esophageal strictures, the rare diagnostic difficulties and the different clinical signs are analyzed. Authors stress that the most frequent causes of the artificial esophageal perforation are the endoscopic removal of swallowed foreign bodies. A surgical intervention in 14 cases was necessary. Three patients were lost after the surgical intervention. PMID- 8430021 TI - [Experience with routine use of combined inhalation-perfusion lung scintigraphy. The most reliable diagnosis of pulmonary embolism]. AB - The authors performed combined inhalation/perfusion scintigraphy in 217 patients with suspected pulmonary embolism in order to improve the diagnostic accuracy. Before the conventional perfusion study they also visualized the distribution of ventilation with a Venticis-II inhalator equipment. Regarding pulmonary embolism 10 (5%), 96 (44%), 39 (18%) investigations were "very low", "low", and "high probability", respectively. The inhalation study was successful in 93 percent of patients. These results were necessary in 141 patients. However, it was uncertain in 26% of cases (57 patients) concerning the diagnosis of pulmonary embolism. As the inhalation scintigraphy was applied without preselection, this study was unnecessary in 30%. On the other hand, if the combined studies had been performed only in the cases of previously proved segmental perfusion defects, the number of perfusion scintigraphies would have increased by 65%. The authors recommend the routine use of the inhalation lung scintigraphy. PMID- 8430023 TI - [The financial structure of the Austrian Nurses Association]. PMID- 8430022 TI - [Rupture of a giant hematoma of the liver as a complication of anticoagulant therapy]. AB - Authors report on a case of a 46-year-old woman, as a rare complication of anticoagulant therapy. Rupture of giant haemangioma of the liver developed after Syncumar treatment. They describe the successful resection of the ruptured tumor, which caused haemorrhagic shock. Based on the review of the literature the authors are the first in Hungary to report on an urgent resection of haemangioma in adult. PMID- 8430024 TI - [Part time nursing personnel--full members of the intensive care team?]. PMID- 8430025 TI - [The nurse's fear of the patient]. PMID- 8430026 TI - [WARD SECRETARY--a new professional branch in the hospital]. PMID- 8430027 TI - [The minister's gifts of words ... homosexuality and society]. PMID- 8430028 TI - [Responsibility for oneself, subordination? Study on the legal situation]. PMID- 8430030 TI - Things I've learned along the way. PMID- 8430029 TI - Death pronouncement by registered nurses. PMID- 8430031 TI - Summary of the NIH Consensus. Triglycerides, high density lipoproteins, and coronary heart disease. PMID- 8430032 TI - An approach to movement disorders of childhood. PMID- 8430033 TI - Myoclonic disorders. PMID- 8430034 TI - Childhood dystonia. PMID- 8430035 TI - Miscellaneous movement disorders of childhood. PMID- 8430036 TI - A pediatrician's view. Stick 'em up and don't move! PMID- 8430037 TI - How organization of nursing care and resident health status affect nursing home costs. PMID- 8430038 TI - Caring for a parent: a phenomenologic inquiry. PMID- 8430039 TI - Self-regulation in the nursing profession: response to substandard practice. PMID- 8430040 TI - Impaired and nonimpaired nurses during childhood and adolescence. PMID- 8430041 TI - Project N.U.R.S.E.: a lesson in empowerment. PMID- 8430042 TI - Toward a theory of technology dependency. PMID- 8430043 TI - Estimates of the supply and demand for doctorally prepared nurses. PMID- 8430044 TI - Behavior of hygroscopic pharmaceutical aerosols and the influence of hydrophobic additives. AB - The high temperature and relative humidity in the lung can result in the hygroscopic growth of susceptible aerosol particles or droplets. The term hygroscopic growth describes the increase in particle diameter which occurs as the result of association with water vapor. The influence of hygroscopicity upon lung deposition of aerosols has been a productive area of research in industrial hygiene, environmental sciences, and inhalation toxicology. Many pharmaceutical inhalation aerosols display hygroscopic behavior in their passage through the airways; however, the effect has been neglected. Controlling the phenomenon of hygroscopic growth and, thus, the related lung deposition of aerosols might result in the therapeutic advantage of targeting the site of action. Such an approach might also allow identification of the location of pharmacologic receptor sites in the lung. This Review discusses an approach to achieving control of hygroscopic growth of aerosol particles. Theoretical and experimental studies have indicated that inhaled particle diameters increased significantly for drugs commonly administered to the lung. The presence of certain additives, notably glycerol, cetyl alcohol, and lauric and capric acids, has been demonstrated to reduce the growth of particles under conditions approaching those in the lung. Very few quantitative studies of the nature discussed herein have appeared in the literature. It is conceivable that an aerosol particle could be fabricated of known initial size and density, and by implication, deposition characteristic, and this might be induced to follow specific growth kinetics to enhance deposition in a particular region of the lung. Thus, physical targeting of regions within the lung might be achieved. PMID- 8430045 TI - Stability of beta-galactosidase, a model protein drug, is related to water mobility as measured by 17O nuclear magnetic resonance (NMR). AB - The inactivation of freeze-dried beta-galactosidase during storage was studied, focusing on the effect of water mobility as measured by the spin-lattice relaxation time, T1, of water using 17O NMR. Inactivation of beta-galactosidase lyophilized from phosphate buffer solution was studied as a function of water content, which in turn affected the T1 of water. An increase in the water content of freeze-dried beta-galactosidase brought about an increase in the T1 of water, as well as a rise in pH. For the freeze-dried enzyme with sufficient water content to be dissolved, the inactivation rate was related to the T1 of water rather than to the pH change. It is suggested that as the water content increases, the mobility of water around the enzyme increases, resulting in enhanced enzyme inactivation. The freeze-dried samples with limited moisture showed inactivation rates faster than those expected from the pH and water mobility, suggesting that the inactivation mechanism is different from that for the freeze-dried enzyme with a larger amount of water. Inactivation of beta galactosidase in solutions was also studied as a function of phosphate buffer and sodium chloride concentrations, which in turn affected the T1 of water. Because the inactivation rate increased with increasing salt concentrations and the rate extrapolated to zero concentration was negligible, inactivation of the freeze dried enzyme was apparently induced by the salts used as additives for lyophilization. The enhancing effect of phosphate buffer components, however, was reduced at higher concentrations, an effect related to the decrease in the T1 of water. This result may be ascribed to the decrease in water mobility caused by phosphate buffer components and is consistent with the observation that the inactivation rate of the freeze-dried enzyme with a relatively large amount of water decreased with decreasing T1 of water. PMID- 8430046 TI - Controlled delivery of pilocarpine. 1. In vitro characterization of Gelfoam matrices. AB - The potential of Gelfoam absorbable gelatin sponge as a carrier for ophthalmic delivery of pilocarpine was examined. Prolonged in vitro release of pilocarpine was achieved through pharmaceutical modification of the device by embedding a retardant in the pores. The device embedded with cetyl ester wax released pilocarpine in a zero-order pattern (release exponent = 0.93 +/- 0.04) for up to 5 hr. This result corresponded well with a linear penetrant uptake by this device. The device impregnated with polyethylene glycol 400 monostearate exhibited anomalous drug transport with a release exponent of 0.63 +/- 0.02. The absorption of water by this retardant and the formation of a gel layer on the surface slowed the penetration of the release medium into the deeper sections of the matrix, as well as the rapid outward diffusion of drug, resulting in a prolonged release of pilocarpine. PMID- 8430047 TI - Comparison of the permeability characteristics of a human colonic epithelial (Caco-2) cell line to colon of rabbit, monkey, and dog intestine and human drug absorption. AB - The in vitro permeabilities of Caco-2 monolayers and permeabilities in tissue sections from colon of monkey, rabbit, and dog were compared using a series of compounds. The selected compounds differed in their physicochemical properties, such as octanol/water partition coefficient, water solubility, and molecular weight. Their structure included steroids, carboxylic acids, xanthins, alcohols, and polyethylene glycols. A linear permeability relationship was established between Caco-2 and colon tissue from both rabbit and monkey. The results suggest that Caco-2 is twice as permeable as rabbit and five times as permeable as monkey colon. However, no clear relationship could be established between Caco-2 monolayers and dog colon permeability. A relationship between permeability in Caco-2 monolayers and human absorption was found. The results suggest that within certain limits, permeability of Caco-2 monolayers may be used as a predictive tool to estimate human drug absorption. PMID- 8430048 TI - Sustained bronchodilation with isoproterenol poly(glycolide-co-lactide) microspheres. AB - An animal study was carried out to evaluate the in vivo bronchodilator action of isoproterenol (Iso) from poly(glycolide-co-lactide) (PGL) microspheres. Microspheres with a mean diameter of 4.5 microns and a drug load of 7% were administered intratracheally to Long-Evans rats. The microspheres released about 70% of the incorporated drug in the instillation medium before administration, which provided immediate action, and the remaining 30% was available for sustained release. A total of 120 animals was anesthetized, paralyzed, artificially ventilated, and divided into 15 groups (n = 8): 3 groups each for saline, blank microspheres, free Iso, blank microspheres with free Iso, and microencapsulated Iso. All instillations were made in a volume of 1 ml/kg and the dose of all Iso preparations was 0.1 mg/kg. At 3, 6, or 12 hr after the intratracheal instillation, a serotonin challenge (40 micrograms/rat) was administered intravenously to constrict the airways. Airway function tests were performed at each time interval on one group of animals by a maximal expiratory flow-volume maneuver. The heart rate in animals receiving Iso formulations was similar to that in the saline control group, indicating minimal systemic effect of the dose administered. The systemic serum levels were below 2 ng/ml in all the groups. Animals receiving encapsulated Iso resisted the serotonin challenge for at least 12 hr after intratracheal instillation, indicating that the drug was still present over this period of time. On the other hand, the serotonin-induced airway constriction observed in the animals receiving blank microspheres, free Iso, or free Iso with blank microspheres was similar to that in saline controls at all time points. The results clearly show that only a small fraction of the free dose is required in sustained-release form for a prolonged pharmacological effect, resulting in a 50- to 100-fold reduction in the total dose administered. PMID- 8430049 TI - Human transbuccal absorption of diclofenac sodium from a prototype hydrogel delivery device. AB - The buccal delivery of the nonsteroidal antiinflammatory drug, diclofenac sodium (Voltaren), from a prototype hydrogel was studied in man in a randomized crossover design of buccal delivery and i.v. infusion. After a 30-min delay, plasma levels of diclofenac increased to near steady-state levels of 100 ng/ml by 3 hr. With each subject serving as his own control, the i.v. infusion data facilitated the calculation of a mean steady-state flux of diclofenac sodium of 2.1 +/- 0.6 mg/cm2-hr across human buccal mucosa and a time lag of 1.0 +/- 0.5 hr. The large flux of this ionized species indicates that the traditional lipoidal model of buccal permeation based on the partition coefficient is inadequate. PMID- 8430050 TI - Relationship among physicochemical properties, skin permeability, and topical activity of the racemic compound and pure enantiomers of a new antifungal. AB - The topical antifungal Sch-39304 is a racemic compound comprised of two enantiomers, Sch-42427 and Sch-42426, only one of which (Sch-42427) is pharmacologically active. The pure enantiomers have a lower melting point and, therefore, a higher solubility than the racemic compound. Because of these differences in physicochemical properties, the concentration of the pure enantiomers in vehicles and in the skin was predicted to be an order of magnitude higher than the racemic compound. It was hoped that the pharmacological activity would also be higher. By measuring the flux of the chiral forms through human cadaver skin, the expected differences in skin solubility were confirmed. However, only a minimal difference between racemate and active enantiomer was observed in the lesion scores using a guinea pig dermatophyte model. By fitting the data to the Emax pharmacodynamic model, it is demonstrated that the maximum effect occurs at a concentration lower than the saturated concentration of the less soluble racemic compound. The data illustrate that the efficacy of topically active compounds may not be linearly related to drug concentration in either the vehicle or the skin. PMID- 8430051 TI - Enantioselective pharmacokinetics of dl-threo-methylphenidate in humans. AB - A definitive enantioselective pharmacokinetic evaluation of dl-threo methylphenidate (MPH) was carried out in 11 healthy volunteers, all of whom received, in a randomized crossover design, three oral administrations of MPH: immediate release (IR), slow release (SR), and SR chewed before swallowing (CH). In addition, all subjects received MPH intravenously (IV) on a separate occasion. Both plasma and urine samples were collected for up to 16 hr after each drug administration. Significant enantioselective differences were found in pharmacokinetic parameters such as CL, MRT, Vdss, AUC infinity 0, and t1/2. A profound distortion of the enantiomeric ratio for MPH (d >> 1) was evident in all plasma samples harvested after oral administration. After IV MPH, however, there was no significant distortion in the plasma d/l ratio until 1.5 hr after dosing, whereafter there was a divergence of the plasma levels of the enantiomers. After oral administration of dl-MPH, the absolute bioavailability (F) of d-MPH was 0.23 and that of l-MPH was 0.05. There were no significant differences in renal clearance for d- or l-MPH after oral or IV administration, although the fraction of the dose excreted unchanged in the urine was significantly greater after IV MPH. These data suggest that enantioselective differences in the pharmacokinetics of oral MPH are the result of enantioselectivity in presystemic metabolism rather than in renal excretion, such that l-MPH is preferentially converted into l ritalinic acid. Finally, it was found that chewing the slow release formulation led to a pharmacokinetic profile very similar to that of MPH-IR, suggesting that MPH-SR should not be prescribed for children who chew tablets. PMID- 8430052 TI - Acacia-gelatin microencapsulated liposomes: preparation, stability, and release of acetylsalicylic acid. AB - Liposomes of dipalmitoylphosphatidylcholine (DPPC) containing acetylsalicylic acid (ASA) have been microencapsulated by acacia-gelatin using the complex coacervation technique as a potential oral drug delivery system. The encapsulation efficiency of ASA was unaltered by the microencapsulation process. The stability of the microencapsulated liposomes in sodium cholate solutions at pH 5.6 was much greater than the corresponding liposomes. The optimum composition and conditions for stability and ASA release were 3.0% acacia-gelatin and a 1- to 2-hr formaldehyde hardening time. Approximately 25% ASA was released in the first 6 hr from microencapsulated liposomes at 23 degrees C and the kinetics followed matrix-controlled release (Q varies; is directly proportional to t1/2). At 37 degrees C, this increased to 75% released in 30 min followed by a slow constant release, likely due to lowering of the phase transition temperature of DPPC by the acacia-gelatin to near 37 degrees C. At both temperatures, the release from control liposomes was even more rapid. Hardening times of 4 hr and an acacia gelatin concentration of 5% resulted in a lower stability of liposomes and a faster release of ASA. It is concluded that under appropriate conditions the microencapsulation of liposomes by acacia-gelatin may increase their potential as an oral drug delivery system. PMID- 8430053 TI - Comparison of the leakage of carboxyfluorescein from symmetric- and asymmetric acyl chain phosphatidylcholine vesicles. PMID- 8430054 TI - Effect of humidity and occlusion on the percutaneous absorption of parathion in vitro. PMID- 8430055 TI - Kinetics of aspirin hydrolysis and stabilization in the presence of 2 hydroxypropyl-beta-cyclodextrin. PMID- 8430056 TI - Pharmacokinetics of stereomeric 1,4:3,6-dianhydrohexitol mononitrates in rats. AB - The pharmacokinetics and urinary recoveries of four isomeric mononitrates, L isoidide mononitrate (L-IIMN), isosorbide-2-mononitrate (IS-2-MN), isomannide mononitrate (IMMN), and isosorbide-5-mononitrate (IS-5-MN), were investigated at an intravenous dose of 2 mg/kg in rats. All four compounds exhibited monoexponential kinetics at this dose. The volumes of distribution were similar for all four isomers and were estimated at about 1.0 liter/kg. The systemic clearances of L-IIMN, IMMN, IS-2-MN, and IS-5-MN were 65.1 +/- 13.0, 32.7 +/- 12.0, 11.0 +/- 2.3, and 8.23 +/- 1.82 ml/min/kg, respectively (P < 0.05, all pairwise comparisons). Free mononitrate in the urine accounted for 0.306 to 4.56% of the administered dose, while the recovery in conjugated forms (after glusulase hydrolysis) accounted for 42.8% of the IMMN dose and 7.70 to 14.5% of the dose of the remaining three isomers. The dose-dependent pharmacokinetics of three of the mononitrates were explored at selected higher doses which cause equivalent vasodilator responses, L-IIMN (22 mg/kg), IS-2-MN (100 mg/kg), and IS-5-MN (300 mg/kg). The clearances of L-IIMN, IS-2-MN, and IS-5-MN at these higher doses were 42.3 +/- 5.7, 6.38 +/- 0.59, and 3.33 +/- 0.62 ml/min/kg, respectively, all significantly less than those found at the 2 mg/kg dose. Typical Michaelis-Menten type curvatures were observed in the concentration-time curves after IS-2-MN and IS-5-MN dosing. The pharmacokinetics of L-IIMN were also dose dependent, but they could not be described by simple Michaelis-Menten kinetics. PMID- 8430057 TI - Steady-state bioavailability and day-to-day variability of a multiple-unit (CR/ZOK) and a single-unit (OROS) delivery system of metoprolol after once-daily dosing. AB - Steady-state bioavailability and day-to-day variability of plasma levels were evaluated in 18 healthy male subjects in a crossover study of multiple once-daily administration of two novel oral drug delivery systems of metoprolol and an immediate-release tablet (100 mg metoprolol tartrate). Data were collected over two consecutive 24-hr dosing intervals on treatment days 6 and 7. The two extended-release formulations investigated were metoprolol CR/ZOK (95 mg metoprolol succinate), a multiple-unit system consisting of several hundred membrane-coated delivery units, and metoprolol OROS (95 mg metoprolol fumarate), a single-unit osmotic delivery system. The extended drug release and absorption observed after administration of metoprolol CR/ZOK and metoprolol OROS resulted in similar steady-state plasma concentrations after once-daily dosing. Compared to the immediate-release tablet, they produced considerably lower plasma peaks, three- to fourfold higher trough concentrations, 8-9 hr longer mean residence times, and 20% lower relative bioavailability. Moreover, the two once-daily metoprolol products were found bioequivalent in Cmax and AUC based on 90% confidence intervals for the mean ratio CR/OROS. Repeated plasma concentration measurements on two consecutive 24-hr periods suggested that all three metoprolol treatments produced reproducible and consistent plasma concentrations from day to day at steady state. Assessment of day-to-day variability, however, resulted in significantly lower variation in AUC for the multiple-unit CR/ZOK formulation compared to the single-unit OROS tablet. These results imply that there may be formulation-related differences in the in vivo behavior of the two products despite their being bioequivalent in extent and rate of absorption. PMID- 8430058 TI - Time-dependent disposition of beta-naphthoflavone in the rat. AB - The pharmacokinetics of beta-naphthoflavone (BNF) have been investigated in rats following various modes of intravenous administration. From intravenous bolus studies it was established that BNF showed a high blood clearance (130 ml/min/kg) and no detectable excretion of unchanged compound in the urine. The volume of distribution for BNF was large (6 L/kg), and binding to plasma proteins extensive (96%). Intravenous infusion studies where the length of infusion was increased from 1 to 8 hr showed marked signs of time-dependent pharmacokinetics. During continuous infusions the plasma concentrations accrued for approximately 1 hr, after which plasma concentrations declined in an apparent exponential fashion to a plateau value. In the short infusion studies the postinfusion half-life (27 min) was significantly shorter than the terminal half-life after bolus administration (40 min). Time-dependent clearance of BNF resulting from enhancement/induction of P450IA enzymes is proposed as the mechanism for these unusual pharmacokinetic features. The use of antipyrine as an independent probe for P450 activity gave similar trends in antipyrine clearance for various modes of BNF administration. Computer simulations based on an autoinduction model for time-dependent clearance were consistent with the observations on BNF in the rat. PMID- 8430059 TI - Distribution of tacrine across the blood-brain barrier in awake, freely moving rats using in vivo microdialysis sampling. AB - Microdialysis was used to sample simultaneously the distribution of THA (9-amino 1,2,3,4-tetrahydroacridine; Tacrine), a potential anti-Alzheimer agent, both in blood and across the blood-brain barrier of anesthetized and awake, freely moving rats. Microdialysis probes were implanted in the jugular vein and dorsal hippocampus and dialysis samples were simultaneously collected from both sites. Dialysis samples were analyzed using a microbore column chromatographic assay with a detection limit of 0.3 ng/ml. Pharmacokinetic parameters were calculated after a 1 mg/kg intravenous dose of THA. Plasma pharmacokinetics followed a biexponential mode, with t1/2(dis.) = 8.4 +/- 2.7 min and t1/2(elim.) = 76.7 +/- 24.2 min for awake, freely moving rats. THA rapidly penetrated the blood-brain barrier, with maximum concentrations attained within 60 min post-dose. In the brain of awake, freely moving rats t1/2(abs.) was 26.0 +/- 5.2 min and t1/2(elim.) was 99.1 +/- 17.7 min. THA levels in hippocampus extracellular fluid were 10 times lower than those in plasma. For anesthetized rats, the t1/2(elim.) in blood was 154.8 +/- 46.8 min, while in the hippocampus t1/2(elim.) was 159.5 +/- 31.7 min. The binding of THA in rat plasma was 56.2 +/- 5.0%, while the fraction bound to rat whole blood was 73.3 +/- 4.1% as determined by microdialysis and ultrafiltration. PMID- 8430060 TI - Prinomide tromethamine pharmacokinetics: mutually dependent saturable and competitive protein binding between prinomide and its own metabolite. AB - Prinomide tromethamine, a nonsteroidal antiinflammatory drug, was orally administered at doses of 250, 500, and 1000 mg every 12 hr for 28 days to healthy male volunteers. The pharmacokinetic behavior of prinomide and its primary plasma metabolite displayed nonlinear characteristics, while those of free prinomide and its metabolite were dose proportional. The nonlinear pharmacokinetic behavior of total prinomide and p-hydroxy metabolite was found to be caused by both saturable and mutually dependent competitive Langmuir-type plasma protein binding between prinomide and its p-hydroxy metabolite. The extent of the protein interaction displayed at steady state was due to the extensive accumulation of the p-hydroxy metabolite. While ligand-protein interactions are known for xenobiotic competitors, the characteristic behavior of prinomide is the first known example to be reported for a competitive protein interaction between a xenobiotic and its own in vivo generated metabolite. The findings of this study may have implications regarding the disposition of other extensively bound nonsteroidal antiinflammatory drugs with long-lived metabolites. PMID- 8430061 TI - Changes in material properties accompanying the national formulary (NF) identity test for microcrystalline cellulose. AB - It has been found that the current NF identity test for microcrystalline cellulose is actually an assay of the response of the material to energy dissipation expended during the analysis procedure. The blending step needed to effect suspension of the material results in a disintegration of the microcrystalline particles and a concomitant increase in the viscosity of the slurry viscosity. These effects were shown not to be unique functions of either blender speed or blending time. The passage or failure of a given sample of microcrystalline cellulose under the conditions of the identity test was found to be a consequence of the test conditions used. Any given sample of microcrystalline cellulose can be made to pass the identity test if it is blended for a sufficiently long time or at a sufficiently high speed. The test is sufficient, however, to differentiate powdered cellulose from powdered cellulose. PMID- 8430062 TI - Prodrugs of peptides. 18. Synthesis and evaluation of various esters of desmopressin (dDAVP). AB - Various aliphatic carboxylic acid esters and a carbonate ester of the tyrosine phenolic group in desmopressin were synthesized to assess their suitability as prodrugs with improved bioavailability compared to the parent peptide. The chemical stability of the esters in aqueous solution was similar to that of simple phenol esters. The derivatives were quantitatively converted to desmopressin by enzymatic hydrolysis in human plasma and rabbit liver homogenate. The esters with a straight side chain were rapidly hydrolyzed by alpha chymotrypsin, but the sterically hindered pivalate ester proved more stable than desmopressin itself toward this proteolytic enzyme. All the esters were more lipophilic than desmopressin in terms of octanol-buffer partition coefficients. The transport of the compounds across confluent monolayers of Caco-2 cells was examined. No correlation between permeability and lipophilicity was found but the pivalate ester showed a markedly higher flux relative to desmopressin. It is concluded that appropriate esterification of desmopressin at its tyrosine group may be a potentially useful prodrug approach. PMID- 8430063 TI - The 3'-keto-diol equilibrium of trospectomycin sulfate bulk drug and freeze-dried formulation: solid-state carbon-13 cross-polarization magic angle spinning (CP/MAS) and high-resolution carbon-13 nuclear magnetic resonance (NMR) spectroscopy studies. AB - Understanding how moisture interacts with a drug or formulation is a critical component of product development. This study demonstrates how water affects the 3'-gem-diol<==>3'-keto equilibrium in trospectomycin sulfate bulk drug and freeze dried formulation, as probed by solid-state carbon-13 cross-polarization magic angle spinning (CP/MAS) and high-resolution nuclear magnetic resonance (NMR) spectroscopy. Drying the bulk drug or formulation to low water levels dehydrates trospectomycin sulfate from the diol to the keto form. Carbon-13 CP/MAS NMR spectroscopy measures the keto drug concentration in solid samples directly. The bulk drug, which contains approximately 16% water, is more than 90% in the 3' diol form. Oven drying to < 3% water converts approximately 75% of the drug to the 3'-keto form. The drug is formulated as a freeze-dried, sterile powder that can contain up to 12% water depending on the freeze-drying conditions. These studies show that the 3'-keto concentration rises uniformly (up to 75%) with decreasing residual water in the freeze-dried cake. The keto-diol equilibrium was also studied in solution by high-resolution carbon-13 NMR experiments, and it was found that raising the temperature or using dimethyl sulfoxide (DMSO) as a solvent also dehydrates the drug. For example, in aqueous solution at 25 degrees C, nearly all (> 95%) of the drug is in the 3'-diol form. After equilibration at 60 degrees C, however, the 3'-keto content increases to 7%, and in d6-DMSO solvent at 25 degrees C the drug is mostly (60%) in the 3'-keto form. PMID- 8430064 TI - Mean residence time of oral drugs undergoing first-pass and linear reversible metabolism. AB - Equations for the mean residence times in the body (MRT) and AUMC/AUC of a drug and its metabolite have been derived for an oral drug undergoing first-pass and linear reversible metabolism. The mean residence times of the drug or interconversion metabolite in the body after oral drug are described by equations which include the mean absorption time (MAT), the mean residence times of the drug or metabolite in the body after intravenous administration of the drug, the fractions of the dose entering the systemic circulation as the parent drug and metabolite, and the systemically available fractions of the drug (Fpp) or metabolite (Fpm). Similarly, the AUMC/AUC of the drug and metabolite after oral drug can be related to the MAT, ratios of the fraction of the dose entering the systemic circulation to the systemically available fraction, the first-time fractional conversion of each compound, and AUMC/AUC ratios after separate intravenous administration of each compound. The Fpp and Fpm values, in turn, are related to the first-pass availabilities of both drug and metabolite and the first-time fractional conversion fractions. The application of these equations to a dual reversible two-compartment model is illustrated by computer simulations. PMID- 8430065 TI - Compression characteristics of granulated materials. VII. The effect of intragranular binder distribution on the compatibility of some lactose granulations. AB - Two sets of lactose-polyvinylpyrrolidone granulations (95:5) of different intragranular binder distributions were produced. The intragranular binder distribution was controlled by a two-step granulation procedure. The compactibility as well as the volume reduction behavior of the granulations was evaluated. Granulations with a more homogeneous distribution of binder in the granules generally produced tablets of a higher mechanical strength than granulations with a peripheral localization of binder. The tablet strength of the latter granulations was also comparatively more reduced by the addition of magnesium stearate. Thus, it is suggested that high granule porosity in combination with homogeneous intragranular binder distribution is advantageous for the compactibility of a granulation. The results of this study therefore contradict earlier suggestions in the literature regarding the preferred intragranular binder distribution. PMID- 8430066 TI - Chemical pathways of peptide degradation. IV. Pathways, kinetics, and mechanism of degradation of an aspartyl residue in a model hexapeptide. AB - In this study the hexapeptide Val-Tyr-Pro-Asp-Gly-Ala (Asp-hexapeptide) was used as a model to investigate the kinetics of aspartate degradation in aqueous solution. The apparent rate of degradation of the Asp-hexapeptide was determined as a function of pH, buffer concentration, and temperature. At very acidic pH levels (0.3, 1.1, 1.5, 2.0, and 3.0), the apparent rate of degradation followed pseudo-first-order kinetics. In this pH region, the Asp-hexapeptide predominantly underwent specific acid-catalyzed hydrolysis of the Asp-Gly amide bond (Asp-X hydrolysis) to form a tetrapeptide (Val-Tyr-Pro-Asp) and a dipeptide (Gly-Ala). In addition, parallel formation of a cyclic imide intermediate could be observed, although no iso-Asp-hexapeptide was detected. At pH 4.0 and 5.0, the Asp hexapeptide simultaneously isomerized via the cyclic imide to form the iso-Asp hexapeptide and underwent Asp-X hydrolysis to produce the cleavage products. The pH-rate profiles (pH 0.3-5.0) for the Asp-X hydrolysis and the formation of cyclic imide revealed that the degree of ionization of the carboxylic acid side chain of Asp residue significantly altered the rate of reaction, with the ionized form being more reactive than the unionized form. Little or no buffer catalysis was observed for either pathway. Solvent isotope experiments were used to probe the mechanism of the Asp-X hydrolysis reaction. At pH values above 6.0, the apparent rate of degradation of the Asp-hexapeptide followed pseudo-first-order reversible kinetics, with the iso-Asp-hexapeptide being the only observed product (isomerization).(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430067 TI - Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C. AB - Although patients with thromboembolic disease frequently have family histories of thrombosis, well-defined defects such as inherited deficiencies of anticoagulant proteins are found only in a minority of cases. Based on the hypothesis that a poor anticoagulant response to activated protein C (APC) would predispose to thrombosis, a set of new coagulation assays was developed that measure the anticoagulant response in plasma to APC. A middle-aged man with a history of multiple thrombotic events was identified. The addition of APC to his plasma did not result in a normal anticoagulant response as measured by prolongation of clotting time in an activated partial thromboplastin time (APTT) assay. Four of the proband's relatives had medical histories of multiple thrombotic events, and they and several other family members responded poorly to APC in the APTT-based assay. Subnormal anticoagulant responses to APC were also found in factor IXa- and Xa-based assays. Several possible mechanisms for the observed phenomenon were ruled out, such as functional protein S deficiency, a protein C-inhibitory antibody, or a fast-acting protease inhibitor against APC. Moreover, restriction fragment-length polymorphism analysis excluded possible linkage of the underlying molecular defect to factor VIII and von Willebrand factor genes. We now describe a previously unrecognized mechanism for familial thromboembolic disease that is characterized by poor anticoagulant response to APC. This would appear to be explained best by a hypothesized inherited deficiency of a previously unrecognized cofactor to APC. As we have identified two additional, unrelated cases with thrombosis and inherited poor anticoagulant response to APC, this may constitute an important cause for familial thrombophilia. PMID- 8430068 TI - Benzo[a]pyrene-induced murine skin tumors exhibit frequent and characteristic G to T mutations in the p53 gene. AB - Human tobacco-related cancers exhibit a high frequency of G to T transversions in the mutation hot spot region of the p53 tumor suppressor gene, possibly the result of specific mutagens in tobacco smoke, most notably benzo[a]pyrene (B[a]P). No in vivo animal model of B[a]P-induced tumorigenesis has been used, however, to substantiate these molecular epidemiological data experimentally. Direct DNA sequence analysis of the hot spot region (exons 5-8 inclusive) of murine p53 was performed in 20 skin tumors induced by a complete carcinogenesis protocol with B[a]P. Sequence analyses revealed numerous heterozygous missense mutations in carcinomas, specifically in exons 7 and 8 of the p53 gene, and targeting exclusively guanine residues. Moreover, 70% (5/7) of the mutations characterized were G to T transversions. In contrast, direct DNA sequence analysis of 36 skin tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) in either a complete carcinogenesis protocol or in a two-stage carcinogenesis protocol revealed a 30% frequency of heterozygous p53 mutations, with the majority of mutations found in carcinomas, but only a single G to T transversion (1/8). Thus, while mutation frequencies are similar, the pattern and type of p53 mutations in B[a]P-induced skin tumors differs significantly from the mutation spectra in DMBA-induced squamous neoplasias. These in vivo findings in B[a]P induced tumors lend support to in vitro and molecular epidemiological evidence, suggesting that the p53 tumor suppressor gene may be a selective target of metabolically activated B[a]P species etiologically associated with human tobacco related cancers. PMID- 8430069 TI - The c-rel protooncogene product represses NF-kappa B p65-mediated transcriptional activation of the long terminal repeat of type 1 human immunodeficiency virus. AB - The long terminal repeat (LTR) of the type 1 human immunodeficiency virus (HIV-1) and the 5' regulatory region of the gene encoding the interleukin 2 receptor alpha subunit (IL-2R alpha) share functional kappa B enhancer elements involved in the regulation of these inducible transcription units during T-cell activation. These kappa B enhancer elements are recognized by a structurally related family of interactive proteins that includes p50, p65, and the product of the c-rel protooncogene (c-Rel). Recent biochemical studies have shown that p65 and p50 form the prototypical NF-kappa B complex, which is rapidly translocated from the cytoplasm to the nucleus during T-cell activation. This intracellular signaling complex potently stimulates kappa B-directed transcription from either the HIV-1 LTR or the IL-2R alpha promoter via the strong transactivation domain present in p65. We now demonstrate that nuclear expression of human c-Rel, which is induced by either phorbol ester or tumor necrosis factor alpha with delayed kinetics relative to p65, markedly represses p65-mediated activation of these transcription units. These inhibitory effects of c-Rel correlate with its DNA binding activity but not with its ability to heterodimerize with p50, suggesting that c-Rel inhibition involves competition with p50/p65 for occupancy of the kappa B enhancer element. Together, these findings suggest that one function of c Rel is as a physiologic repressor of the HIV-1 LTR and IL-2R alpha promoters, serving to efficiently counter the strong transcriptional activating effects of p65. PMID- 8430070 TI - Molecular, functional, and evolutionary analysis of sequences specific to Salmonella. AB - In that salmonellae have been implicated in an unprecedented array of diseases, sequences found to be specific to this species are often thought to be involved in the virulence attributes not seen in other enteric bacteria. To identify the molecular, genetic, and phenotypic characteristics that differentiate bacterial species, we analyzed five cloned DNA fragments that were originally described as being confined to Salmonella. Most of these segments mapped to unique positions on the Salmonella typhimurium chromosome indicative of independent evolutionary events, and three had G+C contents considerably lower than that of the Salmonella genome, suggesting that they arose through horizontal transfer. The nucleotide sequence was determined for one of the clones exhibiting an atypical base composition. This 4.9-kb fragment contained an open reading frame with structural similarity to the LysR family of transcriptional regulators. Strains harboring deletions in this region were tested for > 120 phenotypic characteristics including the effects on a collection of environmentally regulated lac gene fusions. In addition, all deletion strains behaved like the wild-type parent when tested for virulence in mice. PMID- 8430071 TI - A pathway for disulfide bond formation in vivo. AB - Protein disulfide bond formation in Escherichia coli requires the periplasmic protein DsbA. We describe here mutations in the gene for a second protein, DsbB, which is also necessary for disulfide bond formation. Evidence suggests that DsbB may act by reoxidizing DsbA, thereby regenerating its ability to donate its disulfide bond to target proteins. We propose that DsbB, an integral membrane protein, may be involved in transducing redox potential across the cytoplasmic membrane. PMID- 8430072 TI - Modification of antisense phosphodiester oligodeoxynucleotides by a 5' cholesteryl moiety increases cellular association and improves efficacy. AB - Phosphodiester oligodeoxynucleotides bearing a 5' cholesteryl (chol) modification bind to low density lipoprotein (LDL), apparently by partitioning the chol modified oligonucleotides into the lipid layer. Both HL60 cells and primary mouse spleen T and B cells incubated with fluorescently labeled chol-modified oligonucleotide showed substantially increased cellular association by flow cytometry and increased internalization by confocal microscopy compared to an identical molecule not bearing the chol group. Cellular internalization of chol modified oligonucleotide occurred at least partially through the LDL receptor; it was increased in mouse spleen cells by cell culture in lipoprotein-deficient medium and/or lovastatin, and it was decreased by culture in high serum medium. To determine whether chol-modified oligonucleotides are more potent antisense agents, we titered antisense unmodified phosphodiester and chol-modified oligonucleotides targeted against a mouse immunosuppressive protein. Murine spleen cells cultured with 20 microM phosphodiester antisense oligonucleotides had a 2-fold increase in RNA synthesis, indicating the expected lymphocyte activation. Antisense chol-modified oligonucleotides showed an 8-fold increase in relative potency: they caused a 2-fold increase in RNA synthesis at just 2.5 microM. The increased efficacy was blocked by heparin and was further increased by cell culture in 1% (vs. 10%) fetal bovine serum, suggesting that the effect may, at least in part, be mediated via the LDL receptor. Antisense chol-modified oligonucleotides are sequence specific and have increased potency as compared to unmodified oligonucleotides. PMID- 8430073 TI - Escherichia coli cell division protein FtsZ is a guanine nucleotide binding protein. AB - FtsZ is an essential cell division protein in Escherichia coli that forms a ring structure at the division site under cell cycle control. The dynamic nature of the FtsZ ring suggests possible similarities to eukaryotic filament forming proteins such as tubulin. In this study we have determined that FtsZ is a GTP/GDP binding protein with GTPase activity. A short segment of FtsZ is homologous to a segment in tubulin believed to be involved in the interaction between tubulin and guanine nucleotides. A lethal ftsZ mutation, ftsZ3 (Rsa), that leads to an amino acid alteration in this homologous segment decreased GTP binding and hydrolysis, suggesting that interaction with GTP is essential for ftsZ function. PMID- 8430074 TI - Microsatellite mapping of the gene causing weaver disease in cattle will allow the study of an associated quantitative trait locus. AB - A genetic disease in cattle, progressive degenerative myeloencephalopathy (weaver disease), is associated with increased milk production. This association could result from population stratification, from a pleiotropic effect of a single gene, or from linkage disequilibrium between the gene causing weaver disease and a quantitative trait locus (QTL) for milk production. To test these hypotheses, we performed an extensive linkage study in a bovine pedigree segregating for the weaver condition and identified a microsatellite locus (TGLA116) closely linked to the weaver gene (zmax, 8.15; theta, 0.03). TGLA116 and, by extension, the weaver locus were assigned to bovine synteny group 13. This microsatellite can be used to identify weaver carriers, to select against this genetic defect, and to study the effect of the corresponding chromosomal region on milk production in Brown Swiss and other breeds of cattle. PMID- 8430075 TI - Low-frequency chimeric yeast artificial chromosome libraries from flow-sorted human chromosomes 16 and 21. AB - Construction of chromosome-specific yeast artificial chromosome (YAC) libraries from sorted chromosomes was undertaken (i) to eliminate drawbacks associated with first-generation total genomic YAC libraries, such as the high frequency of chimeric YACs, and (ii) to provide an alternative method for generating chromosome-specific YAC libraries in addition to isolating such collections from a total genomic library. Chromosome-specific YAC libraries highly enriched for human chromosomes 16 and 21 were constructed. By maximizing the percentage of fragments with two ligatable ends and performing yeast transformations with less than saturating amounts of DNA in the presence of carrier DNA, YAC libraries with a low percentage of chimeric clones were obtained. The smaller number of YAC clones in these chromosome-specific libraries reduces the effort involved in PCR based screening and allows hybridization methods to be a manageable screening approach. PMID- 8430076 TI - Arachidonic acid status correlates with first year growth in preterm infants. AB - Diets deficient in the omega-6 fatty acid linoleic acid reduce arachidonic acid (Ach) concentrations and retard growth of developing animals and humans. Nevertheless, plasma phosphatidylcholine Ach concentrations declined from 84 +/- 23 mg/liter at birth to a nadir of 38 +/- 11 mg/liter at 4 mo of age in preterm infants fed commercial formulas with linoleic acid, and weight normalized to that of term infants fell progressively beginning at 2 mo of age. The nadir of plasma phosphatidylcholine Ach (31 +/- 7 mg/liter) and growth were further reduced by formula containing marine oil compared with the commercial formulas. Ach status (defined as the mean plasma phosphatidylcholine Ach concentration at 2, 4, and 6.5 mo) correlated with one or more measures of normalized growth through 12 mo. Ach status and maternal height accounted for as much as 59% of the weight variance and 68% of the length variance in infants fed standard formulas. Better Ach status was not from higher energy intakes. A conditional Ach deficiency in preterm infants may contribute to growth over the first year of life. On the strength of the relationship between Ach status and growth, we hypothesize that dietary Ach could improve first year growth of preterm infants. PMID- 8430077 TI - The unusual metal clusters of nitrogenase: structural features revealed by x-ray anomalous diffraction studies of the MoFe protein from Clostridium pasteurianum. AB - Nitrogenase (EC 1.18.6.1) catalyzes the conversion of dinitrogen to ammonia, the central reaction of biological nitrogen fixation. X-ray anomalous diffraction data were analyzed to probe the structures of the metal clusters bound by nitrogenase MoFe protein. In addition to one FeMo cofactor, each half-molecule of MoFe protein binds one large FeS cluster of a type not previously observed in a protein. The FeS cluster contains roughly eight Fe atoms, comprises two subclusters, and is separated from the FeMo cofactor by an edge-to-edge distance of 14 A. The inorganic framework of the FeMo cofactor is not resolved into subclusters, but the Mo atom is located at its periphery. FeMo cofactors in different half-molecules are 70 A apart and cannot promote binuclear activation of dinitrogen by two Mo atoms. PMID- 8430078 TI - Use of photoactivatable peptide substrates of Saccharomyces cerevisiae myristoyl CoA:protein N-myristoyltransferase (Nmt1p) to characterize a myristoyl-CoA-Nmt1p peptide ternary complex and to provide evidence for an ordered reaction mechanism. AB - Nmt1p (EC 2.3.1.97) catalyzes the transfer of myristate (C14:0) from coenzyme A to the N-terminal glycine residue of a variety of eukaryotic cellular and viral proteins. Our recent studies of the 455-amino acid Saccharomyces cerevisiae acyltransferase (Nmt1p) suggested that its mechanism of catalysis is ordered Bi Bi with myristoyl-CoA binding occurring prior to binding of peptide and release of CoA occurring prior to release of the myristoyl-peptide. The interaction between enzyme and peptide has now been examined in greater detail by using photoactivatable octapeptide substrates containing 125I-labeled azidosalicyclic acid attached via an amide bond to the gamma-amino group of a diaminobutyrate residue located at position 2 or the epsilon-amino group of a lysine residue located at position 8. The photopeptides can be specifically crosslinked to chymotryptic fragments of Nmt1p in the presence but not in the absence of a nonhydrolyzable myristoyl-CoA analog, S-(2-oxo)pentadecyl-CoA. Labeling of the chymotryptic fragments is markedly reduced when GLYASKLS, a high-affinity substrate derived from residues 2-9 of S. cerevisiae ADP-ribosylation factor 2, or ALYASKLS, a competitive inhibitor (for peptide), is added with the iodinated photopeptide. These findings suggest that peptide affinity for the acyl-CoA-Nmt1p binary complex is much greater than it is for apoNmt1p, consistent with the ordered Bi Bi mechanism ascribed to Nmt1p. Finally, automated sequential Edman degradation of these chymotryptic fragments suggests that the peptide binding domain of Nmt1p may be composed of elements from two protease-resistant domains, Arg42-Try219 and Thr220-Leu455. PMID- 8430079 TI - Overexpression, purification, and characterization of SHPTP1, a Src homology 2 containing protein-tyrosine-phosphatase. AB - A protein-tyrosine-phosphatase (PTPase; EC 3.1.3.48) containing two Src homology 2 (SH2) domains, SHPTP1, was previously identified in hematopoietic and epithelial cells. By placing the coding sequence of the PTPase behind a bacteriophage T7 promoter, we have overexpressed both the full-length enzyme and a truncated PTPase domain in Escherichia coli. In each case, the soluble enzyme was expressed at levels of 3-4% of total soluble E. coli protein. The recombinant proteins had molecular weights of 63,000 and 45,000 for the full-length protein and the truncated PTPase domain, respectively, as determined by SDS/PAGE. The recombinant enzymes dephosphorylated p-nitrophenyl phosphate, phosphotyrosine, and phosphotyrosyl peptides but not phosphoserine, phosphothreonine, or phosphoseryl peptides. The enzymes showed a strong dependence on pH and ionic strength for their activity, with pH optima of 5.5 and 6.3 for the full-length enzyme and the catalytic domain, respectively, and an optimal NaCl concentration of 250-300 mM. The recombinant PTPases had high Km values for p-nitrophenyl phosphate and exhibited non-Michaelis-Menten kinetics for phosphotyrosyl peptides. PMID- 8430080 TI - SRY recognizes conserved DNA sites in sex-specific promoters. AB - Formation of male-specific structures and regression of female primordia are regulated in early male embryogenesis by SRY, a single-copy gene on the Y chromosome. Assignment of SRY as the testis-determining factor in eutherian mammals is supported by molecular analysis of cytogenetic sex reversal (i.e., XX males and XY females) and by complementary studies of transgenic murine models. Here we characterize the putative DNA-binding domain of SRY, which contains a conserved sequence motif shared by high-mobility group nuclear proteins and a newly recognized class of transcription factors. The SRY DNA-binding domain specifically recognizes with nanomolar affinity proximal upstream elements (designated SRYe) in the promoters of the sex-specific genes encoding P450 aromatase and Mullerian inhibiting substance (MIS). P450 aromatase catalyzes the conversion of testosterone to estradiol, and in the male embryo its expression is down-regulated. Conversely, MIS is expressed in the male embryo to induce testicular differentiation and regression of female reproductive ducts. SRYe binding activity is observed in nuclear extracts obtained from embryonic urogenital ridge immediately preceding morphologic testicular differentiation. Our results support the hypothesis that SRY directly controls male development through sequence-specific regulation of target genes. PMID- 8430081 TI - Copper and iron are mobilized following myocardial ischemia: possible predictive criteria for tissue injury. AB - Direct evidence for substantial mobilization of copper in the coronary flow immediately following prolonged, but not short, cardiac ischemia is presented. In the first coronary flow fraction (CFF) of reperfusion (0.15 ml), after 35 min of ischemia, the level of copper (as well as of iron) was 8- to 9-fold higher than the preischemic value. The levels in subsequent CFFs decreased and reached the preischemic value, indicating that both metals appear in a burst at the resumption of coronary flow. When the first CFF was used in a reaction mixture containing ascorbate and salicylate, the latter underwent chemical hydroxylation and was converted to its dihydroxybenzoate derivatives. Likewise, this CFF promoted the ascorbate-driven DNA degradation. Subsequent 150 CFFs were serially collected and demonstrated low activities. Following 18 min of ischemia, the copper level in the first CFF of reperfusion was only 15% over the preischemic value. In contrast, the mobilization of iron into coronary flow was significant but markedly lower than after 35 min. The levels of copper and the redox activity of the first CFF correlated well with the degree of loss of cardiac function, after 18 and 35 min of ischemia, respectively. After 18 min of ischemia, cardiac function was about 50% and the damage is considered reversible, whereas after 35 min the functional loss exceeded 80% and is considered irreversible. These results are in accord with the causative role that copper and iron can play in heart injury following ischemia, by virtue of their capacity to catalyze the production of hydroxyl radicals, and could lead to the development of new modalities for intervention in tissue injury. PMID- 8430082 TI - Redundant cyclin overexpression and gene amplification in breast cancer cells. AB - Cyclins are prime cell cycle regulators and are central to the control of major check points in eukaryotic cells. The aberrant expressions of two cyclins (i.e., cyclins A and D1) have been observed in some cancers, suggesting they may be involved in loss of growth control. However, in spite of these occasional changes involving only two cyclins, there are no clear connections between general derangements of other cyclins or their dependent kinases in a single tumor type. We detected general cyclin overexpression in 3 of 3 breast tumor tissue samples. In addition, using proliferating normal vs. human tumor breast cells as a model system, we observed a number of alterations in cyclin expression: (i) an 8-fold amplification of cyclin E gene in one tumor line, a 64-fold overexpression of its mRNA, and altered expression of its protein; (ii) deranged expression of cyclin E protein in all (10 of 10) tumor cell lines studied; (iii) increased cyclin mRNA stability, resulting in (iv) general overexpression of RNAs and proteins for cyclins A and B and CDC2 in 9 of 10 tumor lines and (v) deranged order of appearance of cyclins in synchronized tumor vs. normal cells, with mitotic cyclins appearing prior to G1 cyclins. These multiple general derangements in cyclin expression in human breast cancer cells provide evidence linking aberrant cyclin expression to tumorigenesis. PMID- 8430083 TI - Single-stranded shuttle phagemid for mutagenesis studies in mammalian cells: 8 oxoguanine in DNA induces targeted G.C-->T.A transversions in simian kidney cells. AB - A single-stranded shuttle vector has been developed for the purpose of investigating translesional events in mammalian cells. The vector is designed to permit site-specific introduction of defined DNA lesions between a gene for neomycin resistance and its promoter. Efficiencies of translesional synthesis in simian kidney cells (COS) and Escherichia coli are established by determining the number of neomycin- and ampicillin-resistant colonies recovered, respectively, after introduction of a modified vector. Fidelity of translesional synthesis is evaluated by analyzing the nucleotide sequence of progeny phagemid DNA in the region corresponding to the lesion site. This experimental system, capable of detecting mutagenic and nonmutagenic events at and adjacent to the lesion site, was used to establish the mutagenic potential of a single 8-oxoguanine residue in DNA. This modified base, produced by attack of reactive oxygen species on cellular DNA, did not cause a decrease in the number of transformants when single stranded DNA containing the lesion replicated in COS cells or E. coli. The predominant mutations observed (> 78%) were G-->T transversions targeted to the site of the lesion. The mutation frequencies for this event were 2.5-4.8% in COS cells and 1.8% in E. coli. It is concluded that a single-stranded shuttle vector, utilized in conjunction with a site-specific approach, can be used to investigate translesional events in mammalian cells and in bacteria. PMID- 8430084 TI - Studies with glycolysis-deficient cells suggest that production of lactic acid is not the only cause of tumor acidity. AB - Solid tumors have been observed to develop an acidic extracellular environment, which is believed to occur as a result of lactic acid accumulation produced during aerobic and anaerobic glycolysis. Experiments using glycolysis-deficient ras-transfected Chinese hamster lung fibroblasts have been performed to test the hypothesis that lactic acid production within solid tumors is responsible for the development of tumor acidity. The variant cells have defects in glucose transport and in the glycolytic enzyme phosphoglucose isomerase with 1% activity compared to parental cells. Consequently, the in vitro rate of lactic acid production by variant cells was < 4% compared to parental cells. An in vitro correlation between lactic acid production and acidification of exposure medium was observed for parental and variant cells. Implantation of both cell lines into nude mice led to tumors with minimal difference in growth rate. As expected, variant cells died when exposed to hypoxic conditions in culture, and parental tumors were observed to have a larger fraction of cells resistant to radiation due to hypoxia (27%) than variant tumors (2%). Using pH microelectrodes, parental (n = 12) and variant (n = 12) tumors were observed to have extracellular pH (pHe) values of 6.65 +/- 0.07 and 6.78 +/- 0.04 (mean +/- SE, P = 0.13), respectively, whereas normal muscle had a pHe of 7.29 +/- 0.06 (P < 0.0001 for both cell lines). The lactic acid content of variant tumors was found to be similar to that in serum, whereas parental tumors had lactic acid content that was higher than in serum (P < 0.0001). We conclude that there was no correlation between lactic acid content and acidosis for these tumors derived from ras-transfected fibroblasts. These results provide evidence that the production of lactic acid via glycolysis is not the only mechanism responsible for the development of an acidic environment within solid tumors. PMID- 8430085 TI - Glucokinase, glucose sensing, and diabetes. PMID- 8430086 TI - Toward an understanding of the molecular mechanisms of physiological cell death. AB - Cell death is a normal physiological process. Morphological studies have shown that cells that die by physiological mechanisms often undergo characteristic changes termed "apoptosis" or "programmed cell death." Recent work has begun to unravel the molecular mechanisms of these deaths and has shown that one of the primary cell-death pathways is conserved throughout much of evolution. In the nematode Caenorhabditis elegans programmed cell deaths are mediated by a mechanism controlled by the ced-9 gene; in mammals apoptosis can often be inhibited by expression of the bcl-2 gene. The ability of the human BCL2 gene to prevent cell deaths in C. elegans strongly suggests that bcl-2 and ced-9 are homologous genes. Although the process of cell death controlled by bcl-2 can occur in many cell types, there appears to be more than one physiological cell death mechanism. Targets of cytotoxic T cells and cells deprived of growth factor both exhibit changes characteristic of apoptosis, such as DNA degradation. However, bcl-2 expression protects cells from factor withdrawal but fails to prevent cytotoxic T-cell killing. DNA degradation is, thus, not specific for any one cell-death mechanism. The ability of bcl-2 to protect cells from a wide variety of pathological, as well as physiological, stimuli indicates that many triggers can serve to activate the same suicide pathway, even some thought to cause necrosis, and not physiological cell death. PMID- 8430087 TI - Prenyl modification of guanine nucleotide regulatory protein gamma 2 subunits is not required for interaction with the transducin alpha subunit or rhodopsin. AB - Guanine nucleotide-binding regulatory protein (G protein) beta gamma dimers that were active in reconstitution assays were produced in insect cells using the baculovirus/Sf9 insect cell expression system. Sf9 cells were infected either singly or in combination with recombinant baculoviruses containing a human G protein beta 1 gene or a bovine G-protein gamma 2 gene. It was possible to express the beta 1 and gamma 2 gene products independently of each other in this system, as determined by using immunological and metabolic labeling techniques. Further, the ability of recombinant beta and/or gamma chains to function in defined biochemical assays of beta gamma activity was assessed for membrane extracts and supernatant fractions from infected Sf9 cells. Extracts of cells expressing beta or gamma chain alone were inactive in these assays, whereas those from cells coinfected with beta 1 and gamma 2 did display activity. These assays were used to identify recombinant beta gamma dimer migration during chromatographic purification, and the recombinant dimers were purified to near homogeneity. Both the membrane-associated and soluble beta gamma dimers facilitated rhodopsin-catalyzed guanosine 5'-[gamma-thio]triphosphate binding to Gt alpha, the GTP-binding subunit of the retinal G protein transducin (K0.5 of 13 +/- 2 and 36 +/- 5 nM, respectively). Both recombinant beta gamma dimers also facilitated the pertussis toxin-catalyzed ADP-ribosylation of Gt alpha with equal potency (K0.5 of 9 +/- 1 and 10 +/- 3 nM for membrane and soluble dimers, respectively). [3H]Mevalonolactone labeling showed that the gamma 2 subunits of membrane-associated beta gamma dimers incorporated radiolabel, whereas in the soluble form they did not. Thus, prenyl modification of gamma 2 directs the membrane association of the beta 1 gamma 2 dimer and increases its apparent affinity for receptor, but it is not required for the functional interaction(s) of the dimer. PMID- 8430088 TI - Rat acid phosphatase: overexpression of active, secreted enzyme by recombinant baculovirus-infected insect cells, molecular properties, and crystallization. AB - Rat prostatic acid phosphatase (rPAP; orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.2) was expressed in the baculovirus expression vector system. Recombinant protein was secreted into the medium at a high yield by infected insect cells, which were cultured at high density in a 30-liter bioreactor allowing high oxygen content for rapidly growing cells. About 20% of the cell protein produced was rPAP. Partial sequence determination of the N terminus of the purified recombinant secreted protein revealed identity to the native secreted protein, showing that the signal peptide is recognized and properly cleaved in insect cells. The enzyme was purified by using L-(+)-tartrate affinity chromatography. The purified protein had a high specific activity of 2620 mumol.min-1.mg-1 with p-nitrophenyl phosphate at the substrate, and it also showed phosphotyrosine phosphatase activity. The molecular mass of the recombinant rPAP was 155 kDa. Two subunits of 46 kDa and 48 kDa could be detected in SDS/PAGE, but only one subunit of 41 kDa was present after digestion with N glycosidase. The active enzyme is a trimer of subunits differing only in glycosylation. When recombinant rPAP was crystallized with polyethylene glycol 6000 as the precipitant, the crystals were trigonal (space group P3(1)21) with cell dimensions a = 89.4 A and c = 152.0 A. The observed diffraction pattern extends to a resolution of at least 3 A. PMID- 8430089 TI - Crystal structure of an oligonucleotide duplex containing G.G base pairs: influence of mispairing on DNA backbone conformation. AB - The structure of the G.G mispaired dodecanucleotide d(CGCGAATTGGCG)2 has been solved by x-ray crystallography and refined to an R factor of 18.8% at 2.2 A resolution for 3513 reflections. The dodecamer crystallizes as a B-type DNA double helix. It contains two G(anti).G(syn) base pairs--i.e., G-4/G-16(anti).G 21/G-9(syn). The Hoogsteen base pairing involves atoms O-6 and N-7 of the guanine in the syn conformation with atoms N-1 and N-2 of the anti-paired purine. One G.G base pair has a bifurcated hydrogen bond between G-4(N-1)...G-21(N-7) and G-4(N 1)...G-21(O-6). There is little overall structural distortion of the double helix induced as a consequence of the mispairing. The helical width is significantly increased by comparison with the structure of the native duplex, and the minor groove width in the 5'-AATT region is decreased. The G.G base pairing induces high-BII phosphate conformations at residues G-9 and T-20 in addition to more normal BII conformations at G-10 and G-22. It is suggested that these backbone aberrations provide signals for the facile repairability of G.G mispairs in DNA. PMID- 8430090 TI - Exploring the energy landscape in proteins. AB - We present two methods to probe the energy landscape and motions of proteins in the context of molecular dynamics simulations of the helix-forming S-peptide of RNase A and the RNase A-3'-UMP enzyme-product complex. The first method uses the generalized ergodic measure to compute the rate of conformational space sampling. Using the dynamics of nonbonded forces as a means of probing the time scale for ergodicity to be obtained, we argue that even in a relatively short time (< 10 psec) several different conformational substrates are sampled. At longer times, barriers on the order of a few kcal/mol (1 cal = 4.184 J) are involved in the large-scale motion of proteins. We also present an approximate method for evaluating the distribution of barrier heights g(EB) using the instantaneous normal-mode spectra of a protein. For the S-peptide, we show that g(EB) is adequately represented by a Poisson distribution. By comparing with previous work on other systems, we suggest that the statistical characteristics of the energy landscape may be a "universal" feature of all proteins. PMID- 8430091 TI - Transcription of the sex-determining region genes Sry and Zfy in the mouse preimplantation embryo. AB - We have confirmed the faster growth of male preimplantation mouse embryos. We have also studied the transcription of Y chromosomal genes postulated to have a role in sex determination, using the highly sensitive technique of reverse transcription polymerase chain reaction at these early stages. We find that two sex-determining region genes, Sry and Zfy, are transcribed during mouse preimplantation development, while the Zfy homologs Zfx and Zfa and a sex determining region gene originally called A1s9 (now called Ube1y-1) are not. We also show that the anti-Mullerian hormone gene, which contains a Sry consensus binding element in its 5' promoter region, is not transcribed at this time. Developmental curves show that Sry and Zfy are expressed commencing at the two cell stage. These results suggest that mammalian sex determination starts prior to gonad differentiation. PMID- 8430092 TI - Oncostatin M is a mitogen for rabbit vascular smooth muscle cells. AB - The growth regulatory protein oncostatin M was initially discovered in macrophage conditioned medium. We investigated the effects of oncostatin M on cultured rabbit aorta smooth muscle cells (SMCs) and found that the peptide stimulated an increase in the incorporation of [3H]thymidine into DNA. The magnitude of the stimulation was dependent on oncostatin M concentration and SMC confluency. In subconfluent cultures, 1-2 nM stimulated 4- to 5-fold increases in DNA synthesis after 20 hr. Other structurally related cytokines (granulocyte colony-stimulating factor, leukemia inhibitory factor, interleukin 6, ciliary neurotrophic factor) did not affect SMC DNA synthesis. After 5 or 8 days, oncostatin M caused a doubling in SMC number and also induced a transformed phenotype. The combination of oncostatin M and platelet-derived growth factor for 8 days resulted in a 4 fold increase in cell number, approximately the same increase in cell number as induced by the addition of 10% fetal calf serum. Further investigation suggested that the mitogenic effect of oncostatin M was in part due to tyrosine kinase activation. Within 1-2 min, the factor increased phosphotyrosine levels of several SMC proteins. In addition, detectable increases in diacylglycerol levels occurred within 2-5 min, reached 50% above control by 30 min, and remained elevated through 45 min of incubation with oncostatin M. SMC inositol phosphate levels were also elevated within 2 min and then returned to near control values by 20 min. Within 30 min, oncostatin M induced expression of the immediate-early gene EGR-1. These data indicate that oncostatin M may be an important, naturally occurring mitogen for vascular SMCs. PMID- 8430093 TI - Twist and writhe of a DNA loop containing intrinsic bends. AB - The finite-element method of solid mechanics is applied to calculation of the three-dimensional structure of closed circular DNA, modeled as an elastic rod subject to large motions. The results predict the minimum elastic energy conformation of a closed loop of DNA as a function of relaxed equilibrium configuration and linking number (Lk). We apply the method to four different starting states: a straight rod, two rods containing either one or two 20 degrees bends, and a circular O-ring. The results, here at low superhelix density, show the changes in writhe (Wr) and in twist (Tw) as Lk is progressively lowered. The presence of even a single intrinsic bend reduces significantly the linking number change at which Wr first appears, compared to an initially straight, bend-free rod. The presence of two in-phase bends, situated at opposite ends of a diameter, leads to the formation of at least two distinct regions of different but relatively uniform Tw increment. The O-ring begins to writhe immediately upon reduction of Lk, and the Tw increment distribution is sinusoidal along the rod. The mechanics calculations, unlike other theoretical approaches, permit us to calculate Tw and Wr independent of the constraint of constant Lk. PMID- 8430094 TI - Stabilization of alpha-helical structures in short peptides via end capping. AB - The alpha-helix-stabilizing effect of different amino acid residues at the helical termini of short peptides in aqueous solution has been determined. Several dodecapeptides containing alanine, asparagine, aspartate, glutamine, glutamate, and serine at the amino terminus and arginine, lysine, and alanine at the carboxyl terminus were synthesized, and the alpha-helical content of each peptide was measured by using circular dichroism spectroscopy. The trend in alpha helix-inducing ability of these amino acids was found to be as follows: aspartate > asparagine > serine > glutamate > glutamine > alanine at the amino terminus and arginine > lysine > alanine at the carboxyl terminus. Our results agree with the Presta and Rose hypothesis [Presta, L. G. & Rose, G. D. (1988) Science 240, 1632 1641] on the role of end capping in helix stabilization. PMID- 8430095 TI - Convergent transcripts of the yeast PRP38-SMD1 locus encode two essential splicing factors, including the D1 core polypeptide of small nuclear ribonucleoprotein particles. AB - The PRP38 gene of Saccharomyces cerevisiae is necessary for the excision of intron sequences from pre-mRNA and required for the maintenance of maximal levels of U6 small nuclear RNA (snRNA). This report describes the identification of a gene of related function, SMD1, located immediately 3' to PRP38. The PRP38 and SMD1 transcription units are configured in an unusual "tail-to-tail" arrangement with their respective open reading frames terminating on opposite strands of a common 6-bp region. The predicted SMD1 polypeptide, Smd1p, is 40% identical to the D1 protein of human small nuclear ribonucleoprotein particles. Experimentally induced depletion of Smd1p blocks the first step of splicing and results in growth arrest. In addition, the levels of the trimethylguanosine-capped spliceosomal snRNAs, U1, U2, U4, and U5, but not the Prp38p-sensitive U6 snRNA, decrease in response to Smd1p depletion. The cap structures of snRNAs persisting in the absence of SMD1 expression appear to be peculiar, as they are poorly recognized by an anti-trimethylguanosine antibody. These data establish Smd1p as a required component of the cellular splicing apparatus and a factor in snRNA maturation and stability. PMID- 8430096 TI - Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. AB - Apoptosis in embryonic C3H/10T1/2 (clone 8) cells is marked by specific changes in morphology and DNA fragmentation that differ from those found in apoptotic thymocytes. These results demonstrate that ultrastructural changes within the nucleus associated with endonucleolytic degradation are linked with structural degradation at higher levels of chromatin organization. Strand modifications within the internucleosomal linker region are shown to involve alkaline-sensitive sites that appear to be sensitive to S1 endonuclease. Our results suggest that apoptosis is not dependent upon internucleosomal cleavage and may reveal the penultimate step and the nature of the metabolic cascade that leads to cell death. PMID- 8430097 TI - Characterization of downstream elements in a Raf-1 pathway. AB - At the poles of the Drosophila embryo, cell fate is established by a pathway that begins with the activation of a membrane-associated tyrosine kinase (the torso gene product); this then leads to activation of a serine/threonine kinase (Drosophila Raf-1). Activated Raf-1 then leads, by an undefined mechanism, to the transcriptional activation of the tailless (tll) gene; the tll gene product, itself a transcription factor, subsequently regulates the expression of an array of target genes. To further define this pathway, we have utilized sequence comparison between Drosophila melanogaster and Drosophila virilis to identify conserved elements in the tll promoter region. As assessed by DNase I footprinting and promoter dissection experiments, two of these elements are potential regulatory targets of Raf-1-activated transcription factors. Sequence comparison also reveals that the unique residues in the DNA-binding domain of the tll protein, the next component in the pathway, are conserved. One of these residues, the alanine after the last cysteine in the first zinc finger, may be responsible for part of the difference between the tll protein DNA binding site and the closely related half-site of the retinoid/estrogen receptors. Consistent with the rapid turnover of the tll protein, it contains a PEST sequence (rich in proline, glutamate and aspartate, serine, and threonine) that is also conserved. PMID- 8430098 TI - Tumor necrosis factor alpha induces the expression of transforming growth factor alpha and the epidermal growth factor receptor in human pancreatic cancer cells. AB - Recombinant human tumor necrosis factor (TNF)-alpha increased the expression of epidermal growth factor receptor (EGFR) mRNA and protein in all of six human pancreatic carcinoma cell lines tested. In addition, TNF-alpha increased the expression of an EGFR ligand, transforming growth factor (TGF)-alpha, at the mRNA and protein level in all cell lines. Increased expression of EGFR protein was associated with elevated steady-state EGFR mRNA levels. Nuclear run-on analysis showed that increase in EGFR mRNA was due to an increased rate of transcription. Induction of EGFR mRNA expression by TNF-alpha was abrogated by cycloheximide but occurred independently of TNF-alpha-induced production of TGF-alpha protein. Protein kinase A or Gi-type guanine nucleotide-binding proteins were not involved in this process as assessed by using appropriate stimulators and inhibitors of these signal transduction pathways. By contrast, staurosporine, an inhibitor of protein kinase C, partially inhibited, and 4-bromophenacyl bromide, a phospholipase inhibitor, completely inhibited TNF-alpha-dependent EGFR mRNA expression. The phospholipase C-specific inhibitor tricyclodecan-9-yl xanthogenate did not alter TNF-alpha-dependent EGFR mRNA expression, suggesting that phospholipase A2 is involved in the modulation of EGFR expression by TNF alpha. The simultaneous induction of a ligand/receptor system by TNF-alpha suggests that this cytokine modulates autocrine growth-regulatory pathways in pancreatic cancer cells. PMID- 8430099 TI - Protein prenylcysteine analog inhibits agonist-receptor-mediated signal transduction in human platelets. AB - Signal transduction components, including the Ras superfamily of low molecular weight GTP-binding proteins and the gamma subunits of heterotrimeric G proteins, are reversibly carboxyl methylated at C-terminal prenylcysteine residues. We have previously shown that the prenylcysteine analog N-acetyl-S-trans,trans-farnesyl-L cysteine (AFC) inhibits carboxyl methylation of these proteins in human platelets. Here we show that concentrations of AFC that inhibit Ras carboxyl methylation (10-50 microM) also block responses to agonists such as ADP, collagen, arachidonic acid, U46619 (a stable analog of prostaglandin H2), thrombin, and guanosine 5'-[gamma-thio]triphosphate. AFC does not inhibit aggregation induced by effectors such as ionomycin, phorbol 12,13-dibutyrate, and bacterial phospholipase C that bypass G proteins to activate platelets at the level of cytosolic Ca2+ concentration and protein kinase C. These findings indicate that AFC inhibits agonist-receptor-mediated signal transduction in human platelets. PMID- 8430100 TI - Dissociation of human mid-dorsolateral from posterior dorsolateral frontal cortex in memory processing. AB - Work with non-human primates had previously demonstrated that the mid dorsolateral frontal cortex, which comprises cytoarchitectonic areas 46 and 9, plays a critical role in the performance of non-spatial self-ordered working memory tasks, whereas the immediately adjacent posterior dorsolateral frontal cortex (area 8) is critical for the learning and performance of visual conditional associative tasks. The present study used positron emission tomography with magnetic resonance imaging to demonstrate the existence, within the human brain, of these two functionally distinct subdivisions of the lateral frontal cortex. These findings provide direct evidence that, just as the monkey brain, the human lateral frontal cortex is functionally heterogeneous and that comparable anatomical areas underlie similar functions in the two species. PMID- 8430101 TI - Functional activation of the human frontal cortex during the performance of verbal working memory tasks. AB - Regional cerebral blood flow was measured with positron emission tomography during the performance of verbal working memory tasks. The same type of verbal response (i.e., reciting numbers) was required in the control and the two experimental tasks. In the control task, the subjects were required to count aloud. In the two experimental tasks, the subjects were required to maintain within working memory the numbers they generated (self-ordered task) or the numbers generated by the experimenter (externally ordered task). Examination of the difference in activation between these conditions revealed strong bilateral activation within the mid-dorsolateral frontal cortex during both experimental tasks. There was, however, no evidence of additional activation within the mid dorsolateral frontal cortex when monitoring self-generated responses as compared with the monitoring of externally generated responses. These results provide evidence regarding the role of the mid-dorsolateral frontal cortex in mnemonic processing that are in agreement with recent findings from work with non-human primates. PMID- 8430102 TI - Pre-mRNA splicing within an assembled yeast spliceosome requires an RNA-dependent ATPase and ATP hydrolysis. AB - Unlike autocatalyzed self-splicing of group I or group II introns, the removal of pre-mRNA introns in vitro occurs in the spliceosome. The spliceosome is a multicomponent complex composed of pre-mRNA, small nuclear ribonucleoprotein particles, and protein factors. ATP is required for the assembly of the spliceosome and both transesterification reactions. An RNA-dependent ATPase, the product of the yeast PRP2 gene, has been shown to be involved in the first transesterification of pre-mRNA splicing but not in spliceosome assembly. By using ATP analogs, we show that hydrolysis of ATP, mediated through a PRP2 dependent step, is required for the first catalytic event of pre-mRNA splicing. Furthermore, by using a two-step purification procedure, we have isolated a PRP2 containing spliceosome within which the first transesterification readily occurs after the addition of ATP. No additional macromolecules were required. Our results suggest that PRP2 binds to the spliceosome, interacting with an unidentified RNA species in the spliceosome, hydrolyzing ATP and allowing splicing to proceed. We postulate that PRP2 may function to induce a conformational change within the spliceosome. Alternatively, PRP2 may be involved in a proofreading step prior to splicing. PMID- 8430103 TI - sid1, a gene initiating siderophore biosynthesis in Ustilago maydis: molecular characterization, regulation by iron, and role in phytopathogenicity. AB - Iron uptake in Ustilago maydis is mediated by production of extracellular hydroxamate siderophores. L-Or-nithine N5-oxygenase catalyzes hydroxylation of L ornithine, which is the first committed step of ferrichrome and ferrichrome A biosynthesis in U. maydis. We have characterized sid1, a gene coding for this enzyme, by complementation in trans, gene disruption, and DNA sequence analysis. A comparison of genomic DNA and cDNA sequences has shown that the gene is interrupted by three introns. The putative amino acid sequence revealed similarity with Escherichia coli lysine N6-hydroxylase, which catalyzes the hydroxylation of lysine, the first step in biosynthesis of aerobactin. Two transcription initiation points have been determined, both by PCR amplification of the 5' end of the mRNA and by primer extension. A 2.3-kb transcript which accumulates in cells grown under low iron conditions was detected by Northern hybridization. A less abundant 2.7-kb transcript was observed in cells grown in iron-containing medium. By contrast, constitutive accumulation of the 2.3-kb transcript was observed in a mutant carrying a disruption of urbs1, a gene involved in regulation of siderophore biosynthesis. Analysis of the pathogenicity of mutants carrying a null allele of sid1 suggests that the biosynthetic pathway of siderophores does not play an essential role in the infection of maize by U. maydis. PMID- 8430104 TI - Potassium-channel activation in response to low doses of gamma-irradiation involves reactive oxygen intermediates in nonexcitatory cells. AB - Active oxygen species are generated during pathophysiologic conditions such as inflammation and ionizing radiation exposure. We tested the hypothesis that an early cellular event in response to these species involves regulation of ion channels. We exposed cells to gamma-irradiation or treated them with hydrogen peroxide, xanthine/xanthine oxidase, or [3H]thymidine and then monitored channel activity by the technique of whole-cell voltage clamping. Recordings showed that both normal and tumor cells exhibit an increase in K+ currents after treatment with radiation, H2O2, and xanthine/xanthine oxidase but not with high specific activity [3H]thymidine, suggesting that the signal for K+ channel activation originates at the cell membrane. A single noncytotoxic dose of 10 cGy induced measurable levels of K+ currents, suggesting that the induction of currents regulates biochemical changes in response to stress. To test whether channel activity is sensitive to active oxygen species, we pretreated cells with N-acetyl L-cysteine (NAC) to increase cellular pools of free radical scavengers before radiation. In NAC-pretreated cells, K+ channel activation by gamma-irradiation was abolished. It has previously been shown that protein kinase C (PKC) is activated by ionizing radiation and can regulate K+ channels in some cells. However, the effect of radiation on induction of K+ channel activity was independent of PKC, since cells chronically exposed to phorbol esters still produced K+ currents after radiation. These results suggest that an early cellular response to oxidative stress is the activation of K+ channels. PMID- 8430105 TI - Recombinant human sickle hemoglobin expressed in yeast. AB - Sickle hemoglobin has been expressed in the yeast Saccharomyces cerevisiae after site-directed mutagenesis of a plasmid containing normal human alpha- and beta globin genes. Cassette mutagenesis of this plasmid was achieved by inserting a DNA fragment containing the beta-globin gene in the replicative form of M13mp18 to make a point mutation and then reconstituting the original plasmid containing the mutated beta-globin gene. Pure recombinant hemoglobin S was shown to be identical to natural sickle hemoglobin in its ultraviolet and visible absorption bands and by gel electrophoresis, isoelectric focusing, amino acid analysis, mass spectrometry, partial N-terminal sequencing, and functional properties (P50, cooperativity, and response to 2,3-bisphosphoglycerate). In yeast and in mammalian cells, cotranslational processing yields the same N-terminal valine residues of hemoglobin alpha- and beta-chains, but in bacterial expression systems the N terminus is extended by an additional amino acid because the initiator methionine residue is retained. Since the N-terminal valine residues of both chains of hemoglobin S participate in important physiological functions, such as oxygen affinity, interaction with anions, and the Bohr coefficient, the yeast expression system is preferable to the bacterial system for recombinant DNA studies. Hence, mutagenesis employing this expression system should permit definitive assignments of the role of any amino acid side chain in hemoglobin S aggregation and could suggest additional approaches to therapeutic intervention. The engineering of this system for the synthesis of sickle hemoglobin and its purification to homogeneity in a single column procedure are described. PMID- 8430106 TI - Clonal expansions of activated gamma/delta T cells in recent-onset multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic disease characterized by focal demyelination of the white matter of the brain and spinal cord. Central nervous system damage appears to be mediated by infiltrating T lymphocytes and macrophages, and a central role for autoreactive CD4+ T cells has been proposed. However, the initial immune events that lead to the chronic process of MS remain unidentified. We now present evidence that a subset of T lymphocytes bearing gamma/delta T-cell antigen receptors has been activated in patients with recent-onset disease. Cells recovered from the cerebrospinal fluid of subjects with MS were cultured for short periods of time in medium supplemented with T-cell growth factors. Expansions of V delta 1 and V delta 2 T-cell receptor-bearing lymphocytes were found only in cell populations obtained from subjects with recent-onset disease. Similar populations were not expanded in subjects with chronic MS or other neurological diseases. Junctional region sequencing showed the expanded gamma/delta T cells to be oligoclonal in nature, suggestive of specific stimulation by antigen. These results reveal a fundamental difference in the immunopathogenesis of acute vs. chronic disease and provide additional insight into the autoimmune nature of MS. PMID- 8430107 TI - Primary structure of two-chain botrocetin, a von Willebrand factor modulator purified from the venom of Bothrops jararaca. AB - The complete amino acid sequence and location of the disulfide bonds of two-chain botrocetin, which promotes platelet agglutination in the presence of von Willebrand factor, from venom of the snake Bothrops jararaca are presented. Sequences of the alpha and beta subunits were determined by analysis of peptides generated by digestion of the S-pyridylethylated protein with Achromobacter protease I or alpha-chymotrypsin and by chemical cleavage with cyanogen bromide or 2-(2'-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine. Two-chain botrocetin is a heterodimer composed of the alpha subunit (consisting of 133 amino acid residues) and the beta subunit (consisting of 125 amino acid residues) held together by a disulfide bond. Seven disulfide bonds link half-cystine residues 2 to 13, 30 to 128, and 103 to 120 of the alpha subunit; 2 to 13, 30 to 121, and 98 to 113 of the beta subunit; and 80 of the alpha subunit to 75 of the beta subunit. In terms of amino acid sequence and disulfide bond location, two-chain botrocetin is homologous to echinoidin (a sea urchin lectin) and other C-type (Ca(2+)-dependent) lectins. PMID- 8430108 TI - Screening for in vivo protein-protein interactions. AB - We describe an in vivo approach for the isolation of proteins interacting with a protein of interest. The protein of interest is "tagged" with a portion of the biotin carboxylase carrier protein (BCCP), encoded on a specially constructed plasmid, so that it becomes biotinylated in vivo. The "query" proteins (e.g., those in a cDNA library) are tagged by fusing them to the 3' end of the lacZ gene on a lambda vector in such a way that the beta-galactosidase activity is not disrupted. These phage are transfected into cells containing the plasmid encoding the BCCP-tagged protein. The infection lyses the cells and exposes the protein complexes. The BCCP-tagged protein and any associated protein(s) are "captured" by using avidin, streptavidin, or anti-biotin antibody-coated filters. The detection of bound protein is accomplished by directly assaying for beta galactosidase activity on the filters. Positive plaques can be plaque-purified for DNA sequencing. We have tested this approach by using c-Fos and c-Jun as our model system. We show that avidin, streptavidin, or polyclonal anti-biotin (but not a monoclonal anti-biotin) antibody is capable of specifically capturing in vivo biotinylated beta-galactosidase and c-Jun and that this capture is dependent upon the presence of both avidin and the BCCP moiety. Further, complexes containing c-Jun and c-Fos can also be isolated in this manner, and the isolation of this complex is dependent on the presence of c-Fos, c-Jun, avidin, and the BCCP moiety. We discuss the possible uses and limitations of this technique for isolating proteins that interact with a known protein. PMID- 8430109 TI - Two functional soybean genes encoding p34cdc2 protein kinases are regulated by different plant developmental pathways. AB - We have isolated two cDNA clones (cdc2-S5 and cdc2-S6) encoding p34cdc2 protein kinases, homologs of yeast cdc2/CDC28 genes, from a soybean nodule cDNA library. The two sequences share 90% sequence homology in the coding regions. The 5' and 3' noncoding regions are distinct from each other, however, indicating that at least two genes encode p34cdc2 protein kinases in soybean. Both sequences can rescue the cdc28 mutation in Saccharomyces cerevisiae but rescue it with different efficiency. Genomic Southern analysis showed the existence of two copies for each of these genes, which are not closely linked and are nonallelic. The relative expression level of the two soybean p34cdc2 genes varies in different tissues. Expression of cdc2-S5 is higher in roots and root nodules, whereas cdc2-S6 is more actively expressed in aerial tissues, indicating that regulation of these two p34cdc2 genes is coupled with plant developmental pathways. Expression of cdc2-S5 is, furthermore, enhanced after Rhizobium infection, whereas cdc2-S6 fails to respond, suggesting that cdc2-S5 plays a role in nodule initiation and organogenesis. This latter gene preferentially responds to auxin (alpha-naphthaleneacetic acid) treatment, indicating that phytohormones may be involved in the control of cell division mediated by Rhizobium infection. Thus, different p34cdc2 protein kinases may control cell division in different tissues in a multicellular organism and respond to different signals--e.g., phytohormones. PMID- 8430110 TI - Differential patterns of DNA binding by myc and max proteins. AB - c-myc, N-myc, and L-myc genes are subject to highly variable degrees of tissue specific regulation. Their aberrant expression has also been implicated in the pathogenesis of a variety of malignant tumors. The recently identified max protein dimerizes with c-myc to promote its sequence-specific DNA binding. max exists in two forms (long and short) that differ by virtue of a 9-amino acid insertion/deletion at the N terminus. We tested recombinant myc and max proteins for binding to six oligonucleotides containing related c-myc sites. Each myc protein, alone and in association with max proteins, manifested a unique pattern of DNA binding. Phosphorylation of both max proteins was observed when they were incubated in a rabbit reticulocyte lysate. This strongly affected DNA binding by max(long) but not by max(short). Our results point to the existence of specific DNA binding preferences for each of the myc proteins. The 9-amino acid segment that distinguishes max(long) from max(short) appears to serve a regulatory function that provides additional control over DNA sequence recognition. PMID- 8430111 TI - Ribosomal RNA antitermination in vitro: requirement for Nus factors and one or more unidentified cellular components. AB - Using an in vitro transcription assay, we have successfully demonstrated read through of a Rho-dependent terminator by the ribosomal RNA antitermination system. The assay used a DNA template containing a promoter-antiterminator terminator arrangement, RNA polymerase, termination factor Rho, antitermination factors NusA, NusB, NusE, and NusG, and a cellular extract depleted of NusB. Terminator read-through was highly efficient only in the presence of the extract and Nus factors, suggesting that an as yet uncharacterized cellular component is required for ribosomal antitermination. The NusB-depleted extract had no activity in the absence of NusB, confirming an absolute requirement for this protein in ribosomal RNA antitermination. The DNA template requirements were the same as those previously established in vivo; transcription of a wild-type boxA sequence is both necessary and sufficient to promote RNA polymerase modification into a terminator-resistant form. PMID- 8430112 TI - Do all programmed cell deaths occur via apoptosis? AB - During development, large numbers of cells die by a nonpathological process referred to as programmed cell death. In many tissues, dying cells display similar changes in morphology and chromosomal DNA organization, which has been termed apoptosis. Apoptosis is such a widely documented phenomenon that many authors have assumed all programmed cell deaths occur by this process. Two well characterized model systems for programmed cell death are (i) the death of T cells during negative selection in the mouse thymus and (ii) the loss of intersegmental muscles of the moth Manduca sexta at the end of metamorphosis. In this report we compare the patterns of cell death displayed by T cells and the intersegmental muscles and find that they differ in terms of cell-surface morphology, nuclear ultrastructure, DNA fragmentation, and polyubiquitin gene expression. Unlike the T cells, which are known to die via apoptosis, we find that the intersegmental muscles display few of the features that characterize apoptosis. These data suggest that more than one cell death mechanism is used during development. PMID- 8430113 TI - Expression of smooth muscle-specific proteins in myoepithelium and stromal myofibroblasts of normal and malignant human breast tissue. AB - The expression of several differentiation markers in normal human mammary gland myoepithelium and in certain stromal fibroblasts ("myofibroblasts") associated with breast carcinomas was studied by immunofluorescence microscopy of frozen sections. Several antibodies to smooth muscle-specific proteins (smooth muscle alpha-actin, smooth muscle myosin heavy chains, calponin, alpha 1-integrin, and high molecular weight caldesmon) and to epithelial-specific proteins (cytokeratins, E-cadherin, and desmoplakin) were used to show that myoepithelial cells concomitantly express epithelial and smooth muscle markers whereas adjacent luminal cells express only epithelial markers. The same antibodies were used to establish that stromal myofibroblasts exhibit smooth muscle phenotypic properties characterized by the expression of all the smooth muscle markers examined except for high molecular weight caldesmon. In addition, both myoepithelium and myofibroblasts show a significant degree of heterogeneity in smooth muscle protein expression. Thus, myoepithelial cells and stromal myofibroblasts are epithelial and mesenchymal cells, respectively, which coordinately express a set of smooth muscle markers while maintaining their specific original features. The dual nature of myoepithelial cells and the phenotypic transition of fibroblasts to myofibroblasts are examples of the plasticity of the differentiated cell phenotype. PMID- 8430114 TI - Cardiovascular and respiratory effects of tiletamine-zolazepam. AB - The combination of tiletamine and zolazepam is an important dissociative anesthetic-tranquilizer. However, little is known about the effects of this combination on the heart and respiration in rats. Adult, male rats anesthetized with tiletamine-zolazepam alone or tiletamine-zolazepam combined with xylazine or butorphanol were evaluated for changes in heart rate, mean arterial blood pressure, arterial blood pH, and blood gases during a 75-min period of anesthesia. Rats anesthetized with tiletamine-zolazepam had increased mean arterial blood pressure and less respiratory depression than did rats anesthetized with sodium pentobarbital. Tiletamine-zolazepam combined with xylazine at either dose produced bradycardia and a marked hypotension that persisted throughout the 75-min period. This combination produced respiratory depression comparable to tiletamine-zolazepam alone. The addition of butorphanol to tiletamine-zolazepam caused a transient hypotension and bradycardia. Tiletamine-zolazepam plus butorphanol produced a mild to severe respiratory depression that was dose and time dependent. These results demonstrate that: a) Tiletamine-zolazepam is cardiostimulatory, a property consistent with the known cardiovascular effects of other dissociative anesthetics; b) xylazine plus tiletamine-zolazepam is a potent cardiovascular depressant combination; and c) tiletamine-zolazepam plus butorphanol at specific doses is an anesthetic analgesic combination with minimal effects on cardiovascular and respiratory function. PMID- 8430115 TI - Effect of an adenosine A1 agonist injected into substantia nigra on kindling of epileptic seizures and convulsion duration. AB - The substantia nigra pars reticulata (SNr) has been reported to be critically involved in the development and propagation of epileptic seizures, while extracellular adenosine appears to be important for making seizures stop. In the present study, an adenosine A1 receptor agonist [N6-cyclohexyladenosine (CHA); 2.0 nmol/side, or vehicle] was injected bilaterally into the SNr shortly before each of the first five of a series of daily kindling stimuli delivered to the rat amygdala. Injections did not affect the acquisition of kindled afterdischarges or the rate at which seizures developed over subsequent kindling sessions, but convulsions occurring 48-72 h after treatment were significantly shortened. Thus, purinergic mechanisms in the SNr do not appear to be specifically involved in the acquisition of kindled seizures but may contribute to a postictal inhibitory process that shortens the convulsive component. PMID- 8430116 TI - Aversive and antiaversive effects of morphine in the dorsal periaqueductal gray of rats submitted to the elevated plus-maze test. AB - The dorsal periaqueductal gray (DPAG) is a well-known region for processing defensive behavior in the brainstem. Rats implanted with cannulae in the DPAG were submitted to the elevated plus-maze test for 5 min. The effects of morphine following systemic (0.1-1.0 mg/kg) or DPAG administration (5-30 nmol) were compared with the benzodiazepine compound midazolam injected similarly (1-10 mg/kg, IP, and 10-80 nM, DPAG). Morphine and midazolam caused dose-dependent increases in the number of entries and time spent in the open arms. A systemic injection of naloxone in doses that block mu-opioid receptors reversed the effects of centrally administered morphine. Higher doses of morphine (70 nmol) induced a non-naloxone-reversible "fearful" hyperreactivity. It is suggested that low doses of morphine inhibit the neural substrate of aversion in the DPAG, probably through activation of mu-receptors, and that microinjections of higher doses of morphine cause proaversive actions not mediated by these opioid receptors. PMID- 8430117 TI - Effect of propofol on memory in mice. AB - The amnestic effects of the intravenous hypnotic anesthetic agent 2,6 diisopropylphenol (propofol; Diprivan) were studied in a single-trial passive avoidance task. Mice were injected with propofol 10 min before or immediately after training. Memory was impaired in a dose-dependent fashion when the anesthetic was administered before learning, but no amnesia was apparent with posttraining injections. Examination of the acquisition of passive avoidance using a multitrial task showed that propofol-treated mice learned the response normally but forgot the learning significantly faster than vehicle-treated controls. The anterograde amnesia was not the result of state-dependent learning. Propofol also disrupted extinction of fear conditioning when the anesthetic was given during the extinction session. Propofol-induced amnesia could be attenuated by amphetamine (1 mg/kg) injected 30 min before the retention test. PMID- 8430118 TI - Intracerebral haloperidol potentiates the dorsal immobility response in the rat. AB - The effects of intracerebral microinjections of 4-[4-(4-chlorophenyl)-4-hydroxy-1 piperidinyl]-1-(4- fluorophenyl)-1-butanone (haloperidol) (1.0 microgram, 0.5 microliter) in five regions of the brain were tested on the duration of the dorsal immobility response (DIR) and the cling and bar catalepsy in the rat. The duration of the DIR was significantly potentiated (but not the cling and bar catalepsy) following 2-h postinjection of haloperidol in the caudate putamen, nucleus accumbens, and globus pallidus but not in the substantia nigra pars compacta or cortex. These data further expand the previous evidence of regional variations in dopamine to the effects upon inhibitory behaviors. PMID- 8430119 TI - MDL 72222, ketanserin, and methysergide pretreatments fail to alter breaking points on a progressive ratio schedule reinforced by intravenous cocaine. AB - The effects of three serotonin [5-hydroxytryptamine (5-HT)] receptor antagonists on cocaine self-administration behavior were investigated. Specifically, the effects of MDL 72222 (a specific 5-HT3 receptor antagonist), ketanserin (a specific 5HT2 receptor antagonist), and methysergide (an aselective 5-HT1/5-HT2 receptor antagonist) on the breaking points reached by rats on a progressive ratio schedule for cocaine reinforcement were examined. Pretreatments with MDL 72222 (7.5-1,920 micrograms/kg, SC), ketanserin (0.4-6.4 mg/kg, IP), and methysergide (2.5-20 mg/kg, IP) failed to alter breaking points from baseline values. Although tested at twice the highest doses previously reported to have significant behavioral effects, the three 5-HT receptor antagonists were without effect. These data suggest that relatively specific blockade of 5-HT receptor subtypes does not influence the reinforcing effects of cocaine. PMID- 8430120 TI - Time sampling observation procedure for studying drug effects: interaction between d-amphetamine and selective dopamine receptor antagonists in the rat. AB - A rapid time sampling observation procedure combined with two forms of automatic activity assessment is described. The methods are illustrated by examination of the behavioral effects of d-amphetamine, administered to rats either alone or in combination with antagonists selective for D1 or D2 dopamine (DA) receptors. Low doses of d-amphetamine (0.5-4.0 mg/kg) increased photocell counts, rearing, ambulation, and various forms of sniffing. Similar effects were observed in the first 30 min following administration of 8.0 mg/kg d-amphetamine. However, animals exhibited licking, intense sniffing down, and repetitive head and limb movements in the following 30 min; minimal ambulation, rearing, and sniffing up were observed. The "traditional" total photocell count measure did not differentiate between these time-dependent changes in locomotion. On the other hand, latch counts-where subjects had to move between the beams to register a count-adequately demonstrated this change in locomotion. The selective D1 receptor antagonist SCH23390 and selective D2 antagonist raclopride dose dependently inhibited the behavioral changes produced by 1.5 mg/kg d-amphetamine. Higher doses of SCH23390, but not raclopride, produced a behavioral pattern indistinguishable from that observed in control sessions. Both DA antagonists equipotently blocked the intense sniffing down and repetitive head/body movements produced by 8.0 mg/kg d-amphetamine. A narrow intermediate range of doses of SCH23390 reduced the incidences of these behaviors and produced levels of locomotion and sniffing straight that were significantly higher than those observed in control session. This form of behavioral activation was not observed with raclopride. Therefore, this observation procedure revealed subtle differences between the inhibitory effects of SCH23390 and raclopride on d amphetamine-induced behavioral changes. PMID- 8430121 TI - Differentiation of haloperidol and clozapine using a complex operant schedule in the dog. AB - This study aimed to differentiate chronically administered typical (haloperidol) and atypical (clozapine) neuroleptics in the dog using a complex temporal regulation schedule combining operant, voluntary, and involuntary motor parameters. Although clozapine and haloperidol showed some characteristics of neuroleptics, justifying their adherence to the same class of compounds, differences have also been highlighted and compared to the clinical observations. Haloperidol induced catalepsy, tremor, dystony, hyperkinesia, and stereotypy. Subjects produced anticipated responses before any stimulus. Incomplete and delayed responses were also produced. An interpretation in terms of akathisia and anhedonia has been suggested. Clozapine induced tremor, exploration, dystony, and hypersalivation. Subjects produced disinhibitory responses to the negative stimulus and incomplete responses but these latter were submitted to tolerance. The simultaneous presence of tranquilizing and disinhibitory effects has been reported on the clinical potential of clozapine both in cases of positive and negative schizophrenic symptomatologies. PMID- 8430122 TI - Qualitative and quantitative differences in the operant runway behavior of rats working for cocaine and heroin reinforcement. AB - Animals were trained to traverse a straight alley for drug reinforcement consisting of five IV injections of either 0.75 mg/kg/injection cocaine (n = 6) or 0.06 mg/kg/injection heroin (n = 6). Testing involved single daily trials during which the latency to leave the start box and the time required to traverse the alley were recorded for each animal. In addition, input from 12 pairs of infrared photocell detector/emittors placed along the length of the alley provided information on the precise location of the animal at 0.1-s intervals throughout the course of each trial. This information was recorded by computer and provided the basis for construction of graphic representations of each trial in the form of spatiotemporal records that revealed the precise route the subject took in getting to the goal box. The experiment revealed substantial differences in the runway behavior of heroin and cocaine animals. While the heroin group exhibited typical patterns of operant performance in that both start latency and goal times decreased gradually over the course of the experiment, cocaine animals were reliably slower than heroin subjects to leave the start box and exhibited a progressive increase in goal times over trials. The latter effect appeared to be a consequence of a "stop and retreat" behavior that was observed in all six cocaine subjects and increased in frequency as the experiment progressed. Because the runway behaviors exhibited here were emitted prior to delivery of the drug reinforcer, they suggest that the motivational state underlying drug-seeking behavior is qualitatively different for heroin- and cocaine-reinforced animals. PMID- 8430123 TI - Sensitivity of lateral hypothalamic neurons to nicotine: origin and possible correlation with nutritional effects of nicotine. AB - Single-unit activity was recorded extracellularly in the lateral hypothalamus of anesthetized rats. A number of neurons responded to intravenous nicotine, but most failed to respond similarly to local nicotine or systemic administration of a peripheral acting agonist. This finding suggests that these neurons respond indirectly to systemic nicotine through afferent pathways originating in central nicotinoceptive cells. The incidence of response was significantly greater in the cells sensitive to moderate changes in blood glucose. This finding suggests that the effects of peripheral nicotine on food intake and body weight are partly mediated by "glycemia-sensitive neurons" in the lateral hypothalamus. PMID- 8430124 TI - Chronic exposure to lead attenuates cocaine-induced behavioral activation. AB - Adult, male rats were exposed to a diet containing 500 ppm (0.05%) lead for 105 days before testing for cocaine-related changes in activity using a Digiscan activity system. Behavioral testing occurred on 6 successive test days. Activity was recorded for 20 min prior to and 40 min after IP injections of either 10, 20, or 40 mg/kg cocaine HCl, with saline injections on the day preceding each drug test day. Cocaine-induced behavioral activation was evident in control diet animals for all three doses (10, 20, and 40 mg/kg). While 10 mg/kg cocaine HCl did not produce behavioral activation in lead-treated animals, both 20 and 40 mg/kg did result in increased activity comparable to that observed in control counterparts. PMID- 8430125 TI - Influence of beta-casomorphins on apomorphine-induced hyperlocomotion. AB - Derivatives of beta-casomorphin Tyr-Pro-Phe-Pro-Gly and their des-Tyr1 derivatives were investigated on the model of apomorphine-induced hyperlocomotion (1 mg/kg = 3 microM/kg, IP). D-Pip4 CM 5 (5 nM) inhibited the apomorphine hypermotility completely, while D-Phe3 CM 5 (5 nM) and D-Pro4 CM 5 (5 nM) decreased it only to about 50%. The normal exploration was nearly completely inhibited by D-Pro4 CM 5 (40 nM), by D-Pip4 CM 5 (5 nM) depressed to 20%, and by D-Phe3 CM 5 (10 nM) to 35%. The maximum inhibition of apomorphine-induced hyperlocomotion by the des-Tyr-casomorphin derivatives was about 50%. The dose response curves were U-shaped. The exploratory activity was not significantly influenced. The mode of action and the involvement of different neurotransmitter systems in the inhibitory effect of beta-casomorphin derivatives on apomorphine hyperlocomotion are discussed. PMID- 8430126 TI - Gender differences in escape-avoidance behavior of mice after haloperidol administration. AB - Gender differences in the disruptive effects of haloperidol on some reinforced behaviors have been observed in different species. However, the inhibitory action of haloperidol on the acquisition and performance of escape-avoidance behavior has only been investigated in male subjects. The present experiment was designed to investigate possible gender differences in the effects of haloperidol on the initial phase of an escape-avoidance learning task. Male and female mice of the OF1 strain were given a single training session in a shuttle-box. Thirty minutes prior to the behavioral test, mice were injected IP with haloperidol (0.25 mg/kg) or physiological saline (10 ml/kg). Latencies of escape and avoidance responses and the number of nonresponses, escapes, avoidances, pseudoavoidances, crossings during the adaptation period, and crossings during intertrial intervals (ITIs) were evaluated. The disruptive action of haloperidol on the escape-avoidance behavior of the mice was greater in males than in females. The number of nonresponses were higher and the number of escapes lower in treated males than in their female counterparts. These gender differences were not found in control subjects. Activity measures of spontaneous motor behavior (crossings in the adaptation period and during ITIs) did not present gender differences either. Several possible mechanisms responsible for this greater susceptibility of males to the inhibitory effects of haloperidol on escape-avoidance learning are discussed, especially the modulating role of female hormones on dopaminergic activity. PMID- 8430127 TI - The NMDA receptor antagonist MK-801 elicits conditioned place preference in rats. AB - (+)-5-Methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate, (MK 801) a potent noncompetitive antagonist of central NMDA receptors, has been hypothesized to have rewarding properties indicative of abuse potential. To test this hypothesis, the effects of MK-801 on the acquisition of a conditioned place preference and on locomotor activity were assessed and compared with d amphetamine. Both MK-801 (0.03 and 0.1 mg/kg, SC) and d-amphetamine (1.0 mg/kg, SC) administration resulted in the acquisition of a conditioned place preference. However, while both amphetamine and the higher dose of MK-801 produced a behavioral activation during the training period the lower dose of MK-801 did not. These results suggest that MK-801, at doses that produce behavioral activation and below, is rewarding and therefore may have abuse potential. PMID- 8430128 TI - Linopirdine (DuP996) facilitates the retention of avoidance training and improves performance of septal-lesioned rats in the water maze. AB - The behavioral effects of 3,3-bis(4-pyridinylmethyl)-1-phenylindolin-2-one [linopirdine (DuP996)] were investigated on retention of the inhibitory avoidance test in normal mice and acquisition of spatial discrimination in the two-platform water maze task in septal-lesioned rats (a model of cholinergic dysfunction characteristic of Alzheimer's disease). Linopirdine significantly enhanced retention of the inhibitory avoidance response in mice (0.026 mumol/kg) and also reduced the number of errors made by septal-lesioned rats in the water maze to a level comparable to sham-operated animals. At this dose, no effects were observed on septal-lesion-induced hyperactivity in an open field or in unoperated rats tested in the elevated plus-maze anxiolytic test. This study extends previous findings of facilitatory effects of linopirdine on memory and demonstrates improved spatial learning in septal-lesioned rats. As the facilitatory effects on memory are not accompanied by a reduction in the hyperactive state present in septal-lesioned animals, a dissociation between cognitive and noncognitive effects of linopirdine can be differentiated in septal-lesioned rats. PMID- 8430129 TI - Sertraline attenuates hyperphagia in rats following nicotine withdrawal. AB - Chronic nicotine administration can decrease food consumption and body weight. Abrupt withdrawal from nicotine can cause the reverse effect, hyperphagia and rapid weight gain. In the current study, the efficacy of sertraline, a serotonin reuptake inhibitor, on nicotine withdrawal-induced hyperphagia and rapid weight gain was assessed in rats. Sertraline was found to be effective in reversing the increase in feeding that occurred after withdrawal from chronic nicotine administration. Sertraline caused a dose-related decrease in food consumption in control rats not given nicotine. Doses of 5 and 10 mg/kg/day caused significant decreases while 2.5 mg/kg/day caused a slight though nonsignificant decrease in food consumption. Rats in which nicotine was abruptly withdrawn after 3 weeks of administration showed a significant increase in food consumption relative to controls. This increase was eliminated by the high dose of sertraline (10 mg/kg/day), but not by the lower two doses (2.5 and 5 mg/kg/day). Water consumption was affected in a similar fashion. Body weight gain was also affected by sertraline. During the first week after nicotine withdrawal, rats rapidly gained weight, but sertraline attenuated this. The 10-mg/kg dose of sertraline significantly attenuated the nicotine withdrawal-induced weight gain. These results suggest that sertraline can counteract the hyperphagia and rapid weight gain associated with nicotine withdrawal, and might therefore be a useful adjunct to smoking cessation. PMID- 8430130 TI - Alterations in regional brain GABA concentration and turnover during pregnancy. AB - During pregnancy, mice are more susceptible to flurothyl-induced seizures than are nonpregnant control mice. The potential role of brain GABA in mediating this behavior was examined in the present study. GABA concentrations in the cerebellum, hippocampus, striatum, midbrain, and cortex from individual control, pregnant (days 17-18) and delivery-day Heterogeneous Stock mice were assayed using a fluorometric method. Turnover of GABA was assessed by inhibiting metabolism with aminooxyacetic acid and measuring GABA accumulation over the next 2 h. Steady-state GABA concentrations decreased significantly from control in all brain regions during pregnancy. Reductions in GABA concentrations were approximately 25-30% in the affected regions. At parturition, GABA concentrations in the cerebellum and cortex returned to control levels, but hippocampal, striatal, and midbrain GABA levels remained significantly depressed. All the indices of GABA turnover--first-order rate constant, half-life, initial rate of synthesis, and turnover rate (product of first-order rate constant and initial concentration)--showed a significant reduction in pregnancy, which was continued through the time of delivery in all brain regions except the hippocampus. Half life values for GABA increased nearly fourfold in the cerebellum and cortex. These results show that there is a significant alteration in GABAergic systems during pregnancy and parturition. We suggest that the reduction in GABA turnover is a compensatory anticonvulsant mechanism to offset the inherent seizure susceptibility brought about by the reduced level of the major inhibitory neurotransmitter in the brain. PMID- 8430131 TI - Enhancement of the prophagic but not of the antidipsogenic effect of U-50, 488H after chronic amphetamine. AB - Two groups of rats were treated with seven daily injections of either saline or d amphetamine (3 mg/kg IP). On the 2 days following the last injection, rats were tested according to a counterbalanced experimental design, each animal receiving, immediately prior to the beginning of the dark phase, saline on one day and the highly selective kappa-opioid agonist trans- +/- 3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)cyclohexyl]-benzene- acetamide methanesulfonate hydrate [U-50,488H (U50)] on the other. A microcomputer-controlled data acquisition system was used for the structural analysis of the feeding and drinking responses to amphetamine and U50. U50 enhanced feeding and depressed drinking in the first hour. The increased food intake was probably the result of the effect of U50 on the development of satiation and duration of satiety. Chronic amphetamine potentiated the prophagic effect but not the antidipsogenic effect of U50. The structural analysis demonstrated that the characteristics of the prophagic effect of U50 were amplified but not changed. PMID- 8430132 TI - Tissue distribution of two NMDA receptor antagonists, [3H]CGS 19755 and [3H]MK 801, after intrathecal injection in mice. AB - The tissue distribution of [3H]cis-4-phosphonomethyl-2-piperidine carboxylic acid (CGS 19755) and [3H](+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10 imi ne (NK-801) was investigated after a single IT injection into lumbar spinal cord of mice. The level of radioactivity was analyzed in the lumbar, thoracic, and cervical spinal cord, brainstem, frontal cortex, liver, lungs, kidneys, stomach, intestine, spleen, heart, and blood from 5 min up to 6 h after injection. Within the CNS, [3H]CGS 19755 redistributed slowly from the site of injection toward the brainstem and cortex, peaking in the cortex 3-4 h after IT injection. At no time, however, did the relative level per gram of tissue in the frontal cortex exceed 10% of the relative level in the lumbar region of the spinal cord. The highest peripheral level of [3H]CGS 19755 was found in the kidneys. [3H]MK-801 redistributed rapidly from the spinal cord injection site to the peripheral organs. The highest peripheral levels of [3H]MK-801 were found in the lungs and liver, where the radioactivity peaked at 10 and 30-60 min, respectively, after injection. The relative levels of [3H]CGS 19755 were consistently higher in CNS tissues (except for the first 15 min in the frontal cortex) and blood than the corresponding levels of [3H]MK-801. The opposite relationship was true in the liver, lungs, kidneys, stomach, intestine, spleen, and heart. The effect on the response latency in the hot-plate test was quantified in the same animals immediately prior to sacrifice for the distribution study.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430133 TI - Effects of flunarizine on ocular motor and postural compensation following peripheral vestibular deafferentation in the guinea pig. AB - The aim of the present study was to determine if the calcium channel antagonist flunarizine would affect the time course of vestibular compensation for unilateral labyrinthectomy (UL) in guinea pigs. Animals received either a single IP injection of flunarizine 1 h pre-UL or a series of IP injections every 6 h for 24 h post-UL, starting at 6 h post-UL. Flunarizine was dissolved in 50-100% DMSO or suspended in 10% Tween-80 and administered at a dose of 10 mg/kg in the pre-UL condition and 10 or 20 mg/kg in the post-UL condition. All injections were 1 ml/kg in volume. Spontaneous nystagmus (SN), yaw head tilt (YHT), and roll head tilt (RHT) were measured using video analysis. When dissolved in DMSO and administered 1 h pre-UL, 10 mg/kg flunarizine had a small but significant effect on the rate of RHT compensation; otherwise, flunarizine had no significant effects on SN, YHT, or RHT when dissolved in DMSO. When suspended in Tween-80, 10 mg/kg flunarizine pre-UL resulted in a significant decrease in SN frequency and YHT relative to the control group, although the magnitude of the differences was small. When 20 mg/kg was given post-UL, both SN and YHT showed a small but significant change in the rate of compensation. No significant differences in RHT were observed. These results demonstrate that IP administration of flunarizine at a dose of 10-20 mg/kg IP has little effect on vestibular compensation compared to the effects obtained with low IM doses (0.8 mg/kg) of verapamil given 1 h pre-UL. PMID- 8430134 TI - Mitotic activity in expanded human skin. AB - In 20 patients subjected to breast reconstruction by means of tissue expansion, a skin biopsy was obtained at the vertex of the breast before and the day after expansion. Samples were incubated in [3H]thymidine, and the number of labeled cells were counted. A statistically significant rise in the number of labeled basal and suprabasal keratinocytes was seen after expansion. The findings suggest a net gain of tissue not only by stretching but also by formation of new tissue generated by tissue expansion. PMID- 8430135 TI - Increased sensitivity in objective monitoring of tissue expansion. AB - The end point for incremental filling of any tissue expander normally may reliably be assessed using subjective parameters, especially pain, as a guideline. However, in unusual circumstances, such as within denervated areas, the presence of marginal skin circulation, or if rapid expansion is required, an objective monitor may be beneficial to ensure safer expansion. We have previously examined the role of serial measurement of transcutaneous oxygen in this regard. In an effort to refine this sometimes cumbersome approach, a comparison using laser Doppler flowmetry as well has been undertaken in 13 consecutive patients. Both monitoring devices demonstrated a progressive impediment to blood flow with increased expansion. At the pain threshold, both techniques recorded values consistent with adequate skin circulation, thereby proving in general the relative safety of subjective evaluation during the usual course of tissue expansion. Greater sensitivity was noted for the laser Doppler as a monitor, though, via the recording of earlier changes in flow characteristics and more rapid identification of critical thresholds for nonviability. From a more practical standpoint, immediate application without a period for equilibration is another major asset of laser Doppler flowmetry as a system for objective monitoring. PMID- 8430136 TI - Massive teratomas involving the cranial base: treatment and outcome--a two-center report. AB - Massive teratomas involving the cranial base are extremely rare, and to our knowledge, there are no reports of this condition in surviving children. We describe the management of four cases of massive intracranial teratomas, reflecting a combined experience from two major referral centers. We found that when corrected at an early age, the brain parenchyma demonstrated a remarkable ability to reexpand in those children who had appeared to have sustained parenchymal deficits secondary to the mass effect from the tumor. Follow-up thus far suggests that regional skull growth potential may be compromised in some of these patients. Often, the exact pathologic diagnosis of these entities is somewhat different, since they may fall somewhere within the hamartoma heterotopia-teratoma continuum. We believe that the best care of these children is offered through a combined simultaneous craniofacial and neurosurgical approach. PMID- 8430137 TI - The management of septal perforations. AB - Twenty-five patients with septal perforations were seen between July 1, 1983, and June 30, 1989. There were 16 males and 9 females, who ranged in age from 21 to 72 years, with the median age of 41.7 years. The majority of the perforations were traumatic in origin, with 19 cases seen following septal surgery, and were between 1.5 and 2.5 cm (20), with a range of 0.5 to 4 cm. All were located at the posterior border of the quadrilateral cartilage at the vomer-ethmoid junction. All but 3 patients underwent surgical repair, and successful closure was achieved in all but 3. However, only 1 patient remained symptomatic after surgery. The surgical technique employed includes an external rhinoplasty approach, septal and intranasal mucosal flaps, and an autograft of mastoid periosteum or temporalis fascia. PMID- 8430138 TI - Calcitonin gene-related peptide treatment of flaps with compromised circulation in humans. AB - Nineteen patients with surgical or traumatic flaps and one patient with a congenital vein vault deficiency were treated with the vasodilatory compound calcitonin gene-related peptide (CGRP) intravenously. The patients showed compromised circulation due to ischemia and/or venous stasis. In all but three patients, the treatment resulted in an improved tissue circulation, as measured by laser Doppler flowmetry. Thirteen of the flaps survived completely, and in three flaps a partial necrosis was encountered (< or = 25 percent). The three flaps that did not respond at all to calcitonin gene-related peptide treatment did not survive. Calcitonin gene-related peptide was shown to be much more effective than transcutaneous electrical nerve stimulation (TENS). Thus calcitonin gene-related peptide provides a pharmacologic tool to improve blood flow in surgical and traumatic flaps with compromised circulation in humans. PMID- 8430139 TI - Calcitonin gene-related peptide increases the blood flow of port-wine stains and improves continuous-wave dye laser treatment. AB - Ten patients with port-wine stains were treated with continuous-wave dye laser before and after treatment with the vasodilatory compound calcitonin gene-related peptide. The patients were given calcitonin gene-related peptide as an intravenous infusion or as a regional intravenous infusion. Six uniformly colored test sites, each 5 x 10 mm, located within a port-wine stain were selected for clinical evaluation. Laser treatment of these areas was performed on one occasion with three different energy densities: 15, 30, and 45 J/cm2. The port-wine stain circulation was monitored with a laser Doppler flowmeter before and after calcitonin gene-related peptide infusion and immediately after laser treatment. The cosmetic result was evaluated and judged as excellent, desirable, or undesirable. It was shown that calcitonin gene-related peptide is able to induce a long-lasting increase in blood flow in the vascular bed of port-wine stains and that the immediate postlaser laser Doppler flow values may serve as an indication of the outcome of laser therapy. The cosmetic result was often better in the group treated with calcitonin gene-related peptide compared with controls, and an excellent result also was achievable with the use of lower levels of energy. PMID- 8430140 TI - Rigid internal fixation of fractures in the angular region of the mandible: an analysis of factors contributing to different complications. AB - Data relating to 113 patients with 121 mandibular angle fractures treated according to the principles of rigid internal fixation were analyzed to determine which clinical factors are associated with different complications. Certain clinical characteristics were found to be associated with major complications. The use of compression plates seemed to entail disadvantages resulting in some complications. Because of the relatively small cross section of bone surface and particular anatomic features of the angular region, well-adjusted interfragmentary compression is often not possible. A neutral reconstruction plate is considered optimal for rigid osteosynthesis. If a molar tooth in the fracture line has to be extracted, this should be done after fracture stabilization. In most cases, an extraoral approach could not be avoided, but complications associated with this approach were infrequent and well tolerated by patients. PMID- 8430141 TI - Fixation of the vascularized bone graft in mandibular reconstruction. AB - One hundred and forty vascularized bone grafts were used for mandibular reconstruction in 135 patients. Most followed surgical ablation of squamous carcinoma recurring (or persisting) after irradiation. This paper analyzes the fixation techniques and their effects on bony union. Of 140 vascularized bone grafts, 132 survived and were reviewed. Fifty were stabilized using rigid techniques and 82 by interosseous wires. Rigid fixation had a success rate of 94 percent, while nonrigid techniques resulted in bony union in 96.3 percent. The three failed cases of rigid fixation occurred when short mandibular compression plates were used in iliac crest grafts. We conclude that vascularized bone grafts exhibit a high degree of bony union and that fixation technique--rigid versus nonrigid--appears unimportant in this context. Compression plating is clearly bone-dependent. Nevertheless, use of the reconstruction plate simplifies mandibular reconstruction, and the extra hardware does not appear to increase infection or other soft-tissue complications. PMID- 8430142 TI - Endoscopic implant evaluation and capsulotomy. AB - Clinical use of breast endoscopy was begun 5 years ago for the purpose of internal endoscopic capsulotomy. This limited role for the endoscope has been entirely displaced by its use for inspection of implants for leaks or rupture. The materials and equipment are readily available, the proper techniques are safe for patient and implant, the method is easily learned, and the evaluation has been reliable. This report describes our first 50 breast endoscopies and details the technique and the means of learning it. The accuracy is far superior to that of mammogram or ultrasound, and to date, there have been no infections or damaged implants from the technique as described. PMID- 8430143 TI - Hip joint communication with pressure sore: the refractory wound and the role of Girdlestone arthroplasty. AB - Fifteen patients who underwent Girdlestone arthroplasty (proximal femoral head resection) were reviewed at The Johns Hopkins Hospital and Northwestern Memorial Hospital. Ages ranged from 24 to 57 years (mean 36.7 years). All patients were paraplegics or quadriplegics (C7-L3). All patients presented with signs of sepsis and had evidence of osteomyelitis. Soft-tissue reconstruction was most commonly performed with the vastus lateralis, and no femoral stabilization was used. There were no deaths. Recurrent ulcers at the site of the Girdlestone arthoplasty were found in 23 percent of patients in whom follow-up was possible. No recurrence was noted at the original site in 77 percent with a mean follow-up of 20 months. Additional pressure sores occurred at other nonsurgical sites in six patients at a mean of 23.3 months. Girdlestone arthroplasty with soft-tissue coverage is mandatory for successful treatment of pressure sores with hip joint involvement. PMID- 8430144 TI - A new nerve pedicle for finger sensibility: the dorsal digital sensory nerve. AB - Restoration of sensibility to the traumatized finger can be a difficult problem. Two patients with insensibility to the volar distal finger after trauma underwent delayed digital nerve repair. In the first patient, the dorsal branch of the radial proper digital nerve was approximated to the distal stump as a pedicle to span a 12-mm gap resulting from neuroma excision. The second patient had a 14-mm defect after scar-tissue excision 8 months following primary neurorrhaphy after trauma. Reconstruction was performed by approximating the dorsal branch of the radial proper digital nerve to the distal stump. Both patients had fingertip sensibility restored 1 year postoperatively, as documented by two-point discrimination. Anatomic dissections of 12 fresh cadaver fingers revealed a consistent pattern. Of the 24 proper digital nerves dissected, 23 had a distal dorsal sensory branch arising at the midportion of the proximal phalanx. The dorsal branch-vascularized pedicle of the proper digital nerve has not been described previously as a method for restoring finger sensibility in cases not amenable to primary neurorrhaphy. We believe this technique should be added to the repertoire of the practicing hand surgeon. PMID- 8430145 TI - Arterialized venous flap for treating multiple skin defects of the hand. AB - An arterialized venous flap, with arterial blood flowing only through the vein of the flap, was used in each of five patients to cover multiple digital skin defects caused by trauma or burns. Four flaps survived completely, and one showed 30 percent partial necrosis. Although this procedure will require additional refinement, it permits a certain range of motion of the involved digits prior to flap division and inset. PMID- 8430146 TI - Perichondrial arthroplasty in a canine elbow model: comparison of vascularized and nonvascularized techniques. AB - The purpose of this study was to use a canine model to compare the ability of vascularized and nonvascularized perichondrial grafts to resurface an experimental joint defect. In five adult mongrel dogs, a 3 x 2 cm segment of perichondrium, subtended by the intercostal-internal mammary vessels, was harvested and transplanted to a surgically created defect on the ulnar condyle of the humerus. A similar defect in the opposite elbow was repaired with a 3 x 2 cm standard perichondrial graft. Five untreated dogs acted as normal controls. The dogs were allowed to mobilize freely for a period of 6 months. At that time, angiography demonstrated that all microvascular anastomoses were patent. There was no statistical difference in the range of motion of the two elbows, in the radiographic appearance, or in bone density of the two groups of joints. Histologic assessment demonstrated that the vascularized perichondrium formed an articular cartilage with an average thickness of 21.8 microns as compared with 38.5 microns for the nonvascularized perichondrium (p < 0.05). The cartilage in both grafts was morphologically hyaline in type. There were degenerative changes in the grafts with partial separation of the graft from the underlying bone and formation of surface clefts or pits, villous projections, and synovial proliferation. Degenerative changes also were observed on the articular surface opposite the grafted humeral condyle. In two animals the grafts displaced from the recipient condyle, which instead became resurfaced by greater amounts of hyaline-type cartilage with lesser degrees of degeneration and subarticular fibrosis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430147 TI - The dorsal skin-flap model in the rat: factors influencing survival. AB - Several authors have postulated that the standard McFarlane-type dorsal rat flap model can survive as a graft. Therefore, in an effort to better understand the metabolic support governing the survival of this flap, five flap designs on the dorsal surface of the rat were studied. Each was manipulated to control progressively for the metabolic support to the flap by means of skin-graft and/or skin-flap physiology. The flap designs included (1) a standard McFarlane flap (n = 10), (2) a full-thickness "flap" graft (n = 10), (3) a McFarlane flap separated from the bed with plastic sheeting (n = 9), (4) a McFarlane flap separated from the bed by closing the wound beneath the flap (n = 29), and (5) flaps raised as in group 4 after a 2-week delay procedure (n = 9). Based on direct comparisons of both the pattern of necrosis and the surviving surface area in each group, we conclude that (1) the distal aspect of the dorsal rat flap can survive as a graft when in contact with the underlying bed, (2) the "take" of the flap as a graft is variable, and (3) to serve as a reasonable indicator of human flap behavior, the skin-graft effect must be controlled for by separating the flap from the underlying bed. PMID- 8430148 TI - Chronological observation on the energy metabolism of skin flaps by 31P-MRS: a novel approach to evaluate the state of a flap. AB - Although much has been reported on the energy metabolism of various flaps, at present there is no practical noninvasive and nondestructive method to demonstrate chronologically the changes in phosphate compounds in the same subject during the healing process. A skin island flap was obtained from the dorsal side of the posterior extremities in each of 18 Wistar female rats, and a skin arterial island flap (vein included in the pedicle was ligated and divided) also was obtained from the same area in each of 15 Wistar female rats. Chronological changes of ATP, phosphocreatine, and inorganic phosphate were observed by 31P-MRS (magnetic resonance spectroscopy) using surface coils at various intervals ranging from 2 hours to 1 week after the operation. And pH was calculated based on the chemical shift values. The results showed that the PCr/Pi ratio recovered at 4 to 8 hours postoperatively in the survival group, whereas the PCr/Pi ratio lasted at the low level after the operations in the necrosis group. Magnetic resonance spectroscopy is used extensively in studies of brain, heart, and muscle, but we have found only a few papers dealing with magnetic resonance spectroscopy and none with in vivo study in the field of plastic and reconstructive surgery. Since nuclear magnetic resonance imaging is becoming a "bedside" test in the general hospitals, we consider that the PCr/Pi ratio obtained from 31P-MRS can be an effective, noninvasive, and simple method for clinical use in predicting and observing the viability of flaps at an early stage. PMID- 8430149 TI - Pedicle musculocutaneous flap transplantation: prediction of final outcome by transcutaneous oxygen measurements in hyperbaric oxygen. AB - In pedicle musculocutaneous flaps, a local circulatory insufficiency with a total or subtotal ischemia may occur and jeopardize the result of the reconstructive surgery. Transcutaneous oxygen pressure (PtcO2) monitoring has been shown to reflect tissue perfusion and has been advocated to predict the final outcome of ischemic flaps. Unfortunately, under normal atmospheric conditions, this test is not sufficiently discriminative. We evaluate the effect of hyperbaric oxygen conditions on the efficiency of this test. Fifteen patients with pedicle musculocutaneous flap were evaluated by clinical examination and transcutaneous oxygen tension measurements. The final outcome was healing in 7 and failure in 8. In order to determine the predictive value of transcutaneous oxygen tension, measurements were done immediately after the surgical procedure. In ambient air, neither the absolute value of transcutaneous oxygen tension (2.6 +/- 3.6 versus 11.7 +/- 12.6 torr; N.S.) nor the difference or the ratio between the transcutaneous oxygen tension of the flap and the subclavicular reference shows any significant difference according to the outcome (failure or success). The same is true in normobaric oxygen. In hyperbaric oxygen, however, there is a significant difference in transcutaneous oxygen tension between the two groups (12 +/- 12 versus 378 +/- 385 torr; p < 0.01). A transcutaneous oxygen tension higher than 50 torr in hyperbaric oxygen (2.5 atm abs) is the best cutoff value to discriminate success from failure. PMID- 8430150 TI - The effects of heparin and dietary fish oil on embolic events and the microcirculation downstream from a small-artery repair. AB - In recent studies we demonstrated that platelet emboli induced by arterial injury impair the microcirculation. The present study was performed to determine if heparin or dietary cod liver oil reduces the incidence of emboli following the same arterial injury and, in turn, whether these agents prevent the associated decrease in microcirculatory blood flow. The cremaster muscle of 29 male Sprague Dawley rats was isolated on a single neurovascular bundle consisting of the iliac artery and vein and the genitofemoral nerve. Emboli were generated by a thrombogenic injury of the iliac artery, and their number and their subsequent effect on capillary perfusion in the downstream microcirculation were measured. Nine animals received heparin (10-unit IV bolus plus 10 units/hour IV infusion), 10 were fed cod liver oil (10 percent by weight of food) for 3 weeks prior to the experiment, and 9 animals receiving no treatment served as controls. The number of emboli was significantly reduced in the heparin group, but there was no accompanying improvement in capillary perfusion. In contrast, in the cod liver oil group, the number of emboli was not reduced, but there was significant improvement in capillary perfusion. These findings suggest that the harmful effect that platelet emboli have on the microcirculation is probably biochemical in nature (vasoconstriction) rather than related to simple mechanical obstruction to flow. PMID- 8430152 TI - What to do with new bearers of the torch? PMID- 8430151 TI - Pregnancy and plastic surgery residency. PMID- 8430153 TI - Cryoglobulinemia: dilemma for the reconstructive surgeon. AB - Cryoglobulinemia was initially noted to occur predominantly in patients with myeloma, but it is now being detected in a growing number of infectious, collagen vascular, and lymphoproliferative disorders. Two patients with leg ulcers due to cryoglobulinemia are presented. The reconstructive surgeon should consider cryoglobulinemia in the differential diagnosis of skin necrosis that is refractory to conventional therapy, since they may be consulted for wound management. In the vast majority of instances, the patient will be referred with a diagnosis of cryoglobulinemia having already been established. In other circumstances, patients may present to the plastic surgeon with no known history of cryoglobulinemia. The informed reconstructive surgeon can make the diagnosis on the basis of clinical findings. Combination therapy (corticosteroid, immunosuppression, and plasmapheresis) may be of use when areas of skin necrosis, typically in the form of leg ulcers, fail to heal with routine measures. PMID- 8430154 TI - Sclerosing lipogranulomatosis: a case report of scrotal injection of automobile transmission fluid and literature review of subcutaneous injection of oils. AB - For nearly a century, physicians and laypersons have attempted to repair, reconstruct, and embellish the human body in numerous ways by injecting various oils beneath the skin. Soon after Gersuny's first reported subcutaneous injection of oil, the local and systemic complications became apparent. Despite this, the practice of oil injections continues. "Medical grade" silicone injection was investigated in the 1960s to 1980s with varied success and complications. While few physicians practice oil injection therapy, some laypersons continue to subject themselves or their clients to the risk of the disfiguring complications of sclerosing lipogranulomata. Accidental high-pressure injection injury of liquids, so-called grease gun injuries, continues to provide a therapeutic challenge for the hand surgeon. Our case of a man who injected automobile transmission fluid into his scrotum illustrates the classical course and proper management of sclerosing lipogranulomata. A subcutaneous inflammatory and fibrosing reaction occurred with regional lymphadenopathy. The need for complete excision of all involved tissue to treat the condition successfully is illustrated. This case also illustrates the tendency of patients to conceal from their doctors the history of self-injection of foreign bodies. In cases of self injection, psychological counseling might certainly be appropriate. PMID- 8430155 TI - A barrier technique for pulsed irrigation of cavitary wounds. AB - A technique for barrier protection during pulsed irrigation of cavitary wounds is presented. The method is discussed within the context of current concerns regarding intraoperative viral transmission. PMID- 8430156 TI - An intraoperative stent for McIndoe vaginal construction. AB - We report an inexpensive, easy-to-apply vaginal stent for use in McIndoe vaginal reconstruction using Surgi-Stuf inserted in a sterile condom. This material's easy compressibility and good "memory" facilitate good apposition for the skin grafts while being easily placed into the depth of the new vaginal pocket. PMID- 8430157 TI - A self-retaining method to maintain vertical position of the maxilla during Le Fort I osteotomies. AB - A simple technique that relies on a fine wire to maintain vertical relationships of the maxilla during Le Fort I osteotomy is described. The advantages are (1) the hands of the assistant are free to help adapt plates, (2) it decreases the assistant's need to maintain a pose, and (3) the time of the operation decreases. PMID- 8430158 TI - Superglue sealant for persistent leakage of cerebrospinal fluid. AB - A patient is presented in whom a persistent low-pressure CSF leak was stopped by using histoacryl tissue glue following initial local flap closure. PMID- 8430159 TI - Report of the American Society of Plastic and Reconstructive Surgeons' Committee on Physicians' Health. PMID- 8430160 TI - Caring for the Afghan refugees in Peshawar, Pakistan. PMID- 8430161 TI - Mammoscopy: the endoscopic intracapsular evaluation of mammary prostheses. PMID- 8430162 TI - Freud's concept of passivity. PMID- 8430163 TI - CT in the study of antiquities: analysis of a basket-hilted sword relic from a 400-year-old shipwreck. PMID- 8430164 TI - Venous thromboembolic disease: the role of US. AB - Deep vein thrombosis (DVT) is a ubiquitous process that in the acute setting can lead to pulmonary embolism. Chronically, permanent changes that develop within the venous system following an episode of DVT can produce the postphlebitic syndrome, which is associated with pain, swelling, and ulceration. The postphlebitis syndrome can often mimic acute DVT or coexist with it. The clinical evaluation of DVT is ineffective and necessitates a reliable noninvasive diagnostic technique. Compression ultrasound (US) has proved to be the diagnostic method of choice for detection of extremity clot. Femoral and popliteal veins are routinely evaluated for acute clot, but uncertainty exists concerning the need to evaluate the calf veins similarly. US also can be used to diagnose chronic venous changes, which are indicated by the presence of incompetent valves and retrograde blood flow. Upper-extremity venous thrombosis, often induced by indwelling catheters, can also be diagnosed with US. PMID- 8430165 TI - Idiopathic eosinophilic esophagitis: how common is it? PMID- 8430166 TI - Angioplasty, bypass surgery, and amputation for lower extremity peripheral arterial disease in Maryland: a closer look. AB - Tunis and colleagues attempted to assess the effect of peripheral angioplasty in a large population with descriptive epidemiologic methods. Their study suffered from a vague statement of purpose, inappropriate and inadequate outcome measures, undetermined differences in prevalence of peripheral vascular disease and prevalence of risk factors for bypass/amputation in 1989 versus 1979, no differentiation between levels of amputation or between primary and secondary amputation, lack of a unique ICD-9-CM code indicating angioplasty for peripheral vascular disease of the lower extremities, lack of unique patient identifiers, a mistaken perception of the adoption of angioplasty as "widespread" in Maryland, and the assumption of uniform coding accuracy throughout the period of study. We conclude that the study design of Tunis et al was inadequate to determine the beneficial effects of angioplasty or bypass surgery in the treatment of peripheral vascular disease. Moreover, the conclusion by Coffman (2) that "invasive procedures are indicated only for the severely ischemic limb" is completely unsupported by the study data. Physicians should not attempt to apply the results of the study by Tunis et al to individual case situations. It should be further appreciated that the study findings do not provide an adequate basis for policy-making decisions. It is clear that important clinical questions concerning the roles of angioplasty, bypass, and amputation should be answered with more definitive studies. PMID- 8430167 TI - Interventional management of peripheral vascular disease: what did we learn in Maryland and where do we go from here? PMID- 8430168 TI - Lung parenchymal changes secondary to cigarette smoking: pathologic-CT correlations. AB - To determine the histopathologic basis for computed tomographic (CT) interpretation of smokers' lung and the accuracy of CT in the detection of alterations related to cigarette smoking, parenchymal lung lesions were studied from 41 heavy smokers who underwent thoracotomy for removal of a solitary pulmonary nodule. CT scanning of the resected lungs, corresponding exactly to the sections seen on preoperative CT scans, resulted in the following pathologic-CT correlations. Areas of ground-glass attenuation seen on preoperative CT scans (n = 11 [27%]) were related to three main histologic features: (a) accumulation of pigmented macrophages and mucus in the alveolar spaces, associated with mild interstitial inflammation and/or fibrosis (n = 7); (b) thickening of the alveolar walls with inflammatory cells with normal alveolar spaces (n = 3); and (c) presence of organizing alveolitis (n = 1). Parenchymal micronodules depicted presurgically (n = 4 [10%]) corresponded to bronchiolectases with peribronchiolar fibrosis (n = 4) associated with obliterative bronchiolitis in one patient. When emphysema was detected presurgically (n = 21 [51%]), it was always present at pathologic study to a higher extent than initially suspected. PMID- 8430169 TI - Pulmonary tuberculosis: CT findings--early active disease and sequential change with antituberculous therapy. AB - To evaluate findings of active pulmonary tuberculosis on computed tomographic (CT) scans and their sequential changes before and after antituberculous chemotherapy, 29 patients with newly diagnosed pulmonary tuberculosis and 12 patients with recent reactivation were studied prospectively. The diagnosis of active pulmonary tuberculosis was based on positive acid-fast bacilli in sputum (n = 29) and changes on serial radiographs obtained during treatment (n = 12). Twenty-six patients were followed up with CT during treatment for 1-20 months. Lungs from the cadavers of nine other patients, who died of pulmonary tuberculosis, were studied to provide a pathologic basis for diagnosis. At examination with CT, centrilobular lesions (nodules or branching linear structures 2-4 mm in diameter) were most commonly seen (n = 39 [95%]); in the 26 patients with follow-up, most of these lesions disappeared within 5 months after the start of treatment. In 11 of 12 patients with recent reactivation, CT clearly differentiated old fibrotic lesions from new active lesions. Lesions in and around the small airways appear to be the most characteristic CT feature of early active tuberculosis and may be a reliable criterion for disease activity. PMID- 8430170 TI - Neural network analysis of ventilation-perfusion lung scans. AB - A neural network model was constructed to interpret ventilation-perfusion (V/Q) lung scans. This model was trained with data from 100 consecutive V/Q scans with pulmonary angiographic correlation. The network was constructed from 28 input parameters that described various standard V/Q findings, which were fed into a single hidden layer that contained 10-20 nodes. The network output indicated the percentage probability of pulmonary embolism for each set of findings on V/Q scans. This network was then used to classify 28 new scans; the resultant classifications were compared with the rankings of an experienced observer who read the scans without knowledge of the correlative angiographic data. The network with 15 hidden nodes outperformed the experienced observer in prediction of the likelihood of pulmonary embolism in the 28-case test set (P = .039). The neural network has several advantages over current algorithms for interpretation of V/Q scans, including the ability to synthesize many variables into a single conclusion and to learn, or modify itself, at exposure to additional data. PMID- 8430171 TI - CT after reconstructive repair of the sternum and chest wall. AB - Acute mediastinitis and sternal infection after sternotomy are potentially devastating complications, but considerable advances in treatment have been made during the past decade. Sternectomy followed by reconstruction with use of either an omental transposition or a muscle flap has markedly decreased mortality and morbidity. After extensive rib resection, various reconstructive repairs, including the use of polytetrafluoroethylene mesh, have proved successful. The authors retrospectively reviewed 27 postoperative computed tomographic (CT) scans obtained in 19 patients. Twelve of these patients had sternal wounds repaired with either omental or muscle flap procedures. Seven patients had chest wall reconstructions with polytetrafluoroethylene patches, muscle transpositions, or both. The authors found no cases of unexpected or unexplained fluid collections on CT scans obtained beyond the 1st month. Any persistent or recurrent collection is suggestive of infection. If clinical and imaging findings are at odds, imaging directed needle aspiration can help determine whether a fluid collection is infected and in need of further treatment. PMID- 8430172 TI - Negative-pressure pulmonary edema after endotracheal intubation. AB - A review of complications occurring in conjunction with general anesthesia identified eight patients with laryngospasm-induced negative-pressure pulmonary edema after endotracheal intubation. Six male and two female patients (mean age, 31.9 years) developed pulmonary edema immediately or up to 25 minutes after extubation. Radiographs obtained 15-165 minutes after symptoms developed revealed alveolar edema and predominating interstitial edema in four patients each. Seven of eight had bilateral centralized patterns, and one had unilateral pulmonary edema. The mean vascular pedicle width was 67 mm +/- 12, a value 40% higher than the normal mean of 48 mm. The mean cardiothoracic ratio was 0.54 +/- .07 (normal mean, 0.53). There were no radiologic abnormalities of the trachea. Negative pressure pulmonary edema should be suspected in young patients with findings of bilateral centralized pulmonary edema, a wide vascular pedicle, and a normal cardiothoracic ratio in the immediate postoperative period. PMID- 8430173 TI - Polyhedral microcalcifications at mammography: histologic correlation with calcium oxalate. AB - Pathologists have associated calcium oxalate dihydrate (weddellite) in breast biopsy specimens with benign or borderline lesions and rarely with malignancy. These microcalcifications may have a polyhedral shape in histologic specimens owing to their crystalline structure. A retrospective radiologic-histologic correlation study was performed on 300 clusters of microcalcifications to determine if microcalcifications with a polyhedral shape could be found at mammography. In 19 cases (6.3%), polyhedral microcalcifications (PMs) were detected by two radiologists. In 12 of these 19 cases, weddellite crystals were found in the histologic specimen under polarized light; in seven cases, no calcification was found. Although rare, PMs can be found at mammography by radiologists aware of their existence. They are due to weddellite crystals and in this series were associated with benign disease in 89% of cases. Prospective studies are required to determine the frequency of PMs on screening mammograms and to evaluate their negative predictive value. PMID- 8430174 TI - Acute renal obstruction: evaluation with intrarenal duplex Doppler and conventional US. AB - To evaluate duplex Doppler ultrasound (US) in acute renal obstruction, bilateral intrarenal Doppler US was performed in 23 patients with unilateral renal obstruction (proved by means of intravenous urography) of 36 hours duration or less. A mean renal resistive index (RI) was calculated for each obstructed and normal contralateral kidney and compared with findings on conventional US scans. The mean RI in the obstructed kidneys was elevated (.77 +/- .07 [standard deviation]) and was higher than the mean RI in the normal contralateral kidney (.60 +/- .04) (P < .001). RIs in the obstructed kidneys were as follows: .75 or greater in 15 kidneys, .70-.74 (mild RI elevation) in five kidneys (but > or = .10 higher than the RI in the normal contralateral kidney), and less than .70 in three kidneys (two of these three patients had pyelosinus extravasation and one patient had clinical obstruction for only 4-5 hours). RI elevation occurred before collecting-system dilatation in four patients (17%). RI elevation occurs by 6 hours of clinical acute renal obstruction and may precede pyelocaliectasis. Renal duplex Doppler US contributes useful clinical information, especially when US is the first modality used to evaluate acute renal colic. PMID- 8430175 TI - Doppler US assessment of maternal kidneys: analysis of intrarenal resistivity indexes in normal pregnancy and physiologic pelvicaliectasis. AB - Duplex ultrasound was performed in both kidneys of 156 normotensive pregnant women without suspected renal disease and 25 nonpregnant women of childbearing age to determine if resistivity indexes are different between pregnant and nonpregnant women and to assess whether the "physiologic" pelvicaliectasis of pregnancy causes elevated resistivity indexes. The mean intrarenal resistivity index for pregnant patients was 61.1, and that for nonpregnant women was 61.0. There were no statistically significant differences between the resistivity indexes of pregnant and nonpregnant women, regardless of whether the pregnant patients were considered as a composite group or subdivided by trimester. Twenty nine right kidneys and four left kidneys in pregnant patients had moderate dilation of the collecting system attributed to physiologic pelvicaliectasis of pregnancy. No statistically significant differences were detected between the resistivity indexes of kidneys with and without pelvicaliectasis. Normal, uncomplicated pregnancy has no significant effect on resistivity indexes obtained in maternal kidneys. Physiologic pelvicaliectasis of pregnancy is likely to be partially a result of obstruction; this too, however, is not reflected by changes in resistivity indexes. An elevated resistivity index during pregnancy should not be attributed to the normal physiologic changes of pregnancy. PMID- 8430176 TI - Renal oncocytoma and carcinoma: failure of differentiation with CT. AB - The authors studied the hypothesis that oncocytoma and adenocarcinoma of the kidney can be differentiated with computed tomographic (CT) criteria and that differences would become more apparent as tumors enlarged. On contrast material enhanced scans, homogeneous attenuation throughout the tumor and a central, sharply marginated, stellate area of low attenuation were considered predictors of oncocytoma. Any area of decreased attenuation in the tumor except for a stellate, central area was used as a predictor of adenocarcinoma. Among oncocytomas larger than 3 cm in diameter, 67% exhibited the criteria for oncocytoma and 33% met the criterion for adenocarcinoma; among smaller oncocytomas, the respective results were 82% and 18%. Among adenocarcinomas larger than 3 cm in diameter, 84% fulfilled the criterion for malignancy and 16% were incorrectly predicted to be oncocytomas; among smaller adenocarcinomas, the respective results were 58% and 42%. The authors conclude that the CT criteria used are poor predictors of the diagnosis of oncocytoma or adenocarcinoma regardless of tumor size. PMID- 8430177 TI - Adnexal masses: comparison of specificity of endovaginal US and pelvic MR imaging. AB - In a prospective study, 32 women with suspected pelvic masses at physical examination underwent both endovaginal ultrasound (US) and magnetic resonance (MR) imaging to compare their ability in diagnosis of adnexal masses. Criteria for the diagnosis of various types of adnexal masses with MR imaging and endovaginal US were prospectively defined, and the ability of either modality to allow a specific diagnosis was assessed. For each modality, measures of sensitivity, specificity, and accuracy were obtained. Results indicated higher diagnostic capability of endovaginal US for simple cysts (five of five), hemorrhagic cysts (eight of nine), endometriomas (nine of 14), and ovarian carcinomas (three of three). MR imaging demonstrated higher diagnostic capability for dermoids (three of three). MR imaging and endovaginal US showed equal diagnostic capability for pedunculated fibroids (two of two). For all masses, observers, and observations, the overall sensitivity of endovaginal US was 76% versus 49% for MR imaging, and the overall accuracy of endovaginal US was 83% versus 70% for MR imaging. The authors suggest that endovaginal US is a better modality than MR imaging for the assessment of suspected pelvic masses. PMID- 8430178 TI - Adrenal masses: differentiation with chemical shift, fast low-angle shot MR imaging. AB - The purpose of this study was to investigate the efficacy of chemical shift, fast low-angle shot (FLASH) imaging at 1.5 T to differentiate adrenal masses. The materials included patients with adrenocortical adenomas (n = 32), metastatic tumors (n = 17), and pheochromocytomas (n = 4). FLASH images were obtained with breath holding at 100/11 (repetition time msec/echo time msec) (out-of-phase images) and 100/13 (in-phase images) and a flip angle of 20 degrees to differentiate the lipid contents in the adrenal tumors. The signal-intensity (SI) indexes of adrenal masses ([SI on IP - SI on OP]/[SI on IP x 100]), where IP = in phase image and OP = out-of-phase image, were calculated. All adenomas had SI indexes of more than 5%, while SI indexes of metastatic tumors and pheochromocytomas were less than 5%, with accuracy of 100% in the differentiation between adenoma and nonadenoma. Hyperfunctioning adenoma, however, could not be differentiated from nonhyperfunctioning adenoma. Chemical shift FLASH imaging was superior to the calculated T2 in the characterization of adrenal masses. PMID- 8430179 TI - Pheochromocytomas that do not accumulate metaiodobenzylguanidine: localization with PET and administration of FDG. AB - Many imaging methods can be used to detect pheochromocytoma, but some tumors are not detected with conventional modalities. To explore the possible usefulness of positron emission tomography (PET) after administration of 2-[fluorine-18]-fluoro 2-deoxy-D-glucose (FDG) to localize pheochromocytoma in patients with false negative scintigrams obtained after administration of metaiodobenzylguanidine (MIBG), FDG was administered and PET was performed in two adult patients with pheochromocytomas that had never been localized despite administration of MIBG. In both patients, images were obtained dynamically for 50 minutes; then a limited truncal sequence was performed. PET enabled correct localization of the tumors. In patient 1, a tumor that had not been detected for 21 years was localized in the middle mediastinum; in patient 2, a pheochromocytoma was detected in the right adrenal gland. PET performed after administration of FDG may be useful for localization of pheochromocytomas that do not accumulate MIBG. PMID- 8430180 TI - Balloon dilation of ureteral strictures after renal transplantation. AB - Ten ureteral strictures that developed in nine patients after renal transplantation were managed with balloon catheter dilation and placement of a ureteral stent. Four strictures were successfully dilated (40%), with a follow-up of 15-42 months (mean, 29 months). Comparison of these cases with the six cases of unsuccessfully dilated strictures failed to show any substantial differences between the groups with respect to demographics, stricture characteristics, or radiologic management techniques. However, strictures that developed at the ureteroneocystostomy site responded favorably more often (three of four strictures) to balloon catheter dilation than did strictures in other locations. The authors' experience is not as favorable as that of others who have managed renal transplant-related ureteral strictures in a similar manner. Nonetheless, their experience reinforces the efficacy of balloon catheter dilation of ureteral strictures that develop after renal transplantation as an effective alternative to surgical revision in a substantial percentage of patients. PMID- 8430181 TI - Temporomandibular joint: diagnostic accuracy with sagittal and coronal MR imaging. AB - To determine the diagnostic accuracy, sensitivity, and specificity of magnetic resonance (MR) imaging of the temporomandibular joint (TMJ) in the assessment of disk position, disk form, and changes in the osseous components, 55 fresh cadaver joints were imaged. A 1.5-T imager was used in the sagittal and coronal planes. After MR imaging, the joints were cryosectioned in the coronal plane corresponding to the coronal image through the center of the joint. The joints were then remounted and sectioned serially in the sagittal plane from lateral to medial, corresponding to the sagittal MR images. MR images were interpreted by the two authors together, without knowledge about cryosectional findings. The cryosections were interpreted by the authors together, without knowledge of the MR imaging findings. MR imaging was 95% accurate in the assessment of disk position and disk form and 93% accurate in the assessment of osseous changes. Coronal images helped avoid a false-negative diagnosis in 13% (n = 7) of the joints. MR imaging with a surface coil appears to be an accurate method for the assessment of soft and hard tissues of the TMJ. PMID- 8430182 TI - Proton MR spectroscopy of HIV-infected patients: characterization of abnormalities with imaging and clinical correlation. AB - The purpose of this study was to characterize the proton magnetic resonance (MR) spectroscopic findings in the brains of patients infected with human immunodeficiency virus (HIV). Peak areas were used to calculate metabolite ratios. Spectra were analyzed by blinded readers who calculated areas under metabolite peaks. MR images were evaluated by blinded readers, with both white and gray matter being rated as normal or abnormal. An aggregate index that combined N-acetylaspartate/creatine (Cr), choline/Cr, and marker peak/Cr ratios resulted in mean scores for patients and control subjects of 4.4 +/- 1.5 (standard deviation [SD]) and 2.5 +/- 0.4, respectively (P = .001). Eight of 11 patients (73%) had abnormal MR images versus four of 11 control subjects. Thirteen of 15 patient spectra (87%) were abnormal (> 2 SDs from the mean of the control subjects), while only one of 10 control spectra was abnormal. These initial results indicate that proton MR spectroscopy is a potentially useful modality for detecting HIV involvement in the central nervous system. PMID- 8430183 TI - Human brain tumors: assessment with in vivo proton MR spectroscopy. AB - To better understand variations in spectra of brain tumors, 122 in vivo proton spectra of brain tumors in 82 patients were analyzed. The changes in relative metabolite concentrations compared with those in normal spectra and the presence of any new metabolite were assessed. To evaluate the clinical usefulness of in vivo hydrogen-1 magnetic resonance (MR) spectroscopy in brain tumors, the authors looked for specific spectral changes on the basis of tumor grade. All tumor spectra showed differences from normal reference spectra. The differential diagnosis of the spectra was limited because intraindividual differences between spectra of one tumor at different locations were often larger than differences between spectra of tumors with different histologic characteristics. However, the variations in metabolite concentrations, and especially the presence or absence of aliphatic signals, were proved to be indicators of the histologic grade of tumor. The observed spectral patterns conformed to a four-compartment model, described herein, which is proposed to improve the interpretation of brain spectra. PMID- 8430184 TI - Intracranial MR angiography: application of magnetization transfer contrast and fat saturation to short gradient-echo, velocity-compensated sequences. AB - To circumvent artifacts related to saturation, turbulence, and data processing in three-dimensional (3D) time-of-flight (TOF) magnetic resonance (MR) angiography, the authors used a high-resolution (256 x 512), short, asymmetric velocity compensated gradient-echo sequence, magnetization transfer contrast (MTC) between all excitation pulses to suppress brain parenchyma signal, and frequency-offset fat saturation (FS) to suppress fat signal. Signal intensity and blood-background contrast was measured in several regions of interest on conventional 3D TOF MR images, MTC images, and those with MTC and FS in seven volunteers. One patient each with an arteriovenous malformation, an aneurysm, and stenosis in the intracranial vascular system underwent the protocols with an echo time (TE) of 5 msec and then a TE of 8 msec. Use of the 5-msec-TE sequence with MTC and FS led to reduced problems associated with signal void in areas of fast flow in patients. In volunteers and patients, vascular visualization was superior to that of current 3D TOF techniques. PMID- 8430185 TI - Unenhanced emergency cranial CT: optimizing patient selection with univariate and multivariate analyses. AB - Charts from 1,074 consecutive emergency department patients who underwent cranial computed tomography (CT) were reviewed for predictors of a CT abnormality. Twenty six clinical variables and the results of neurologic examination were compared with cranial CT findings. Patients with focal neurologic deficit, unresponsiveness, and hypertension had an increased risk of a CT abnormality. Blurred vision, trauma, loss of consciousness, headache, and dizziness were each associated with a lower risk of a CT abnormality. Multivariate analysis showed that only focal neurologic deficit and unresponsiveness effectively helped predict a CT abnormality. In patients with negative neurologic findings, only intoxication and amnesia were associated with greater than 10% positive scans and an increased risk for a CT abnormality. The data indicate that positive neurologic findings coupled with intoxication and amnesia would have helped detect 90.7% of the positive scans and provide an effective initial approximation strategy for selecting patients to undergo CT. Although 15 patients with positive scans (1.4%) would have been missed, this strategy would have yielded a negative predictive value of 97.3% and eliminated 53.9% of the CT scans obtained. PMID- 8430186 TI - Phase III multicenter clinical investigation to determine the safety and efficacy of gadoteridol in children suspected of having neurologic disease. AB - A phase III open-label clinical trial was conducted at 11 institutions to determine the safety and efficacy of gadoteridol in children suspected of having neurologic disease. One hundred three children were included in the safety analysis; 92 were evaluated for efficacy (76 intracranial and 16 spinal examinations). Three adverse events were reported in two children. All adverse events were considered minor and resolved spontaneously without treatment or sequelae. In a comparison of enhanced T1-weighted magnetic resonance images with unenhanced T1- and T2-weighted images, enhancement of disease was noted in 70% of the intracranial and 38% of the spinal examinations. Additional diagnostic information was reported in 82% of the postcontrast intracranial examinations and 62% of the spinal examinations. Use of this additional information contributed to a potential modification of patient diagnosis in 48% of intracranial and 20% of spinal cases with additional information. These results indicate excellent safety and efficacy for use of gadoteridol in children with suspected intracranial or spinal disease. PMID- 8430187 TI - Pediatric intracranial vascular malformations: evaluation of treatment results with color Doppler US. Work in progress. AB - To determine the value of color Doppler ultrasound (US) in the preoperative assessment and evaluation of treatment results in children with intracranial vascular malformations (VMs), the authors performed 36 color Doppler US studies in nine children who underwent endovascular embolization. Color flow imaging with spectral waveform analysis of feeding arteries, nidus, draining veins, and uninvolved cranial vasculature was performed, and correlation was made with other imaging findings. In most VMs, color Doppler US enabled the authors to map the lesion completely. Hemodynamic changes after embolization included improvement in blood supply to uninvolved portions of the brain and increase in caliber and flow of feeding vessels that were not occluded during embolization. Serial volume flow measurements were performed with Doppler US in major extracranial arteries. Success of embolization was indicated by substantial decrease of total carotid artery flow. Color Doppler US is a noninvasive modality that adds important imaging and hemodynamic data to those provided by angiography. PMID- 8430188 TI - Multicystic dysplastic kidney in children: US follow-up. AB - Eighty-one cases of multicystic dysplastic kidney (MCDK) in children were diagnosed over the past 11 years at the authors' institution: 25 children had their kidneys surgically removed, eight with bilateral total involvement died, and 48 underwent serial follow-up ultrasonography (US) of their kidneys. Follow up included 193 serial ultrasound (US) studies (mean, four per patient) for a total of 1,468 months (mean, 30.5 months). In the 48 patients followed up, 32 (67%) kidneys showed a decrease in size, nine (19%) showed no change, and five (10%) increased in size, and in two (4%), a change in size could not be determined. In seven of the 48 (15%) children, the MCDKs decreased in size, and, at follow-up US, no renal tissue could be found. In those patients in whom MCDKs decreased in size. Serial US characteristics changed from predominantly an enlarged cystic structure to a small dysplastic or absent kidney. Two of the five kidneys that increased in size were surgically removed, and MCDK was pathologically confirmed. A nonsurgical approach to the treatment of patients with MCDK is supported by this study. PMID- 8430189 TI - Idiopathic eosinophilic esophagitis. AB - A retrospective review was performed of 13 patients with the diagnosis of idiopathic eosinophilic esophagitis (IEE) occurring alone or in conjunction with idiopathic eosinophilic gastroenteritis (IEG) to identify clinical, radiographic, endoscopic, manometric, and therapeutic similarities. All patients presented with esophageal symptoms, predominantly dysphagia. An allergic disorder was present in 10 (77%) patients, and peripheral eosinophilia was present in 12 (92%) patients. Clinical investigation disclosed esophageal strictures in 10 patients, motility disorders in three, ulcerations in two, a cervical web in one, and a mucosal ring in one as the cause of esophageal symptoms. A proximal esophageal stricture was the single most common esophageal abnormality demonstrated. IEE should be included in the differential diagnosis of dysphagia and should be suggested in a patient with an allergic disorder, peripheral eosinophilia, and concurrent abdominal symptoms, especially in conjunction with IEG. Prompt diagnosis is extremely important since treatment with steroids produces rapid clinical remission in most patients. PMID- 8430190 TI - Mn-DPDP in MR imaging of pancreatic adenocarcinoma: initial clinical experience. AB - Manganese (II) N,N'-dipyridoxylethylenediamine-N,N'-diacetate 5,5'-bis(phosphate) (DPDP) was tested as a contrast agent for magnetic resonance imaging of pancreatic adenocarcinoma in 15 patients. At enhanced T1-weighted spin-echo (SE) and enhanced T1-weighted gradient-echo (GRE) imaging, statistically significant increases in signal-to-noise ratio (S/N) for the pancreas (21% and 92%, respectively) and contrast-to-noise ratio (C/N) (91% and 209%, respectively) were found. The C/N at enhanced T1-weighted SE imaging was superior to that at unenhanced imaging, including T2-weighted SE imaging (P = .001). Subjective image analysis showed that delineation of the pancreas and pancreatic tumors was clearly improved (P = .05) on enhanced T1-weighted SE and GRE images compared with on unenhanced T1- and T2-weighted images. The liver enhanced 19% at T1 weighted SE imaging and 90% at T1-weighted GRE imaging. There was a significantly higher S/N increase in hepatic parenchyma than in pancreatic tissue at enhanced T1-weighted GRE imaging (P = .0026) but not at enhanced T1-weighted SE imaging. PMID- 8430191 TI - Abdominal MR imaging: comparison of T2-weighted fast and conventional spin-echo, and contrast-enhanced fast multiplanar spoiled gradient-recalled imaging. AB - T2-weighted fast and conventional spin-echo (SE) and dynamic gadolinium-enhanced fast multiplanar spoiled gradient-recalled (FMPSPGR) images in 26 patients (18 with hepatic masses and eight with no detected abnormality) were compared to determine the efficacy of the newer pulse sequences (fast SE and FMPSPGR) in abdominal imaging. FMPSPGR allows for breath-hold magnetic resonance imaging of the liver. The contrast-to-noise ratio (C/N) of the hepatic lesions was calculated for each sequence and was superior in fast SE compared with that in conventional SE in 16 of 18 patients. Lesion detection was 90% with fast SE and was 85% and 80% for conventional SE and contrast material-enhanced FMPSPGR sequences, respectively. Of the T2-weighted sequences, fast SE showed consistently sharper anatomic structures and less respiratory and cardiac motion artifact. Thus, fast SE (with its superior C/N and shorter imaging time) and gadolinium-enhanced FMPSPGR images, when combined, demonstrate potential value as routine sequences in abdominal imaging. PMID- 8430192 TI - Cartilaginous tumors: correlation of gadolinium-enhanced MR imaging and histopathologic findings. AB - Gadolinium-enhanced magnetic resonance (MR) images were correlated with histopathologic findings in patients with cartilaginous tumors. Short repetition time/echo time spin-echo MR images obtained before and after intravenous administration of gadopentetate dimeglumine in 34 patients were evaluated. Twenty four of 27 low-grade chondrosarcomas showed septal enhancement. The enhancing curvilinear speta consisted of fibrovascular tissue. The nonehancing areas consisted of paucicellular hyaline cartilage, cystic mucoid tissue, and necrosis. This enhancement pattern and corresponding histopathologic findings were not seen in three benign osteochondromas or four high-grade chondrosarcomas. Inhomogeneous or homogeneous enhancement correlating with highly cellular areas at histopathologic analysis was seen in all high-grade chondrosarcomas. The three osteochondromas showed peripheral enhancement that correlated with fibrovascular tissue covering the non-enhancing cartilage cap. The authors conclude that septal enhancement on MR images after intravenous administration of gadopentetate dimeglumine improves tissue characterization of cartilaginous tumors and may assist in identifying low-grade chondrosarcoma. PMID- 8430193 TI - Extraskeletal mesenchymal chondrosarcoma. AB - Among seven patients with extraskeletal mesenchymal chondrosarcoma (EMC), three children (aged 3-6 years) developed EMC in a central location and four adults (aged 38-54 years) developed EMC in both central and peripheral sites. Conventional radiography and tomography and computed tomography (CT) depicted EMC as a soft-tissue mass with ring, arc, stippled, and highly opaque calcifications in four patients. Contrast-enhanced CT showed lobulation and peripheral tumoral enhancement, sometimes with central low-attenuation areas. On magnetic resonance (MR) images, EMC was a lobulated mass with high signal intensity on T2-weighted images and enhancement with low-signal-intensity focal areas on contrast-enhanced T1-weighted images. All adults developed recurrences and/or metastases and died. Of the three children, two were living and free of disease at the end of the study and the third child died of chemotherapeutic-induced leukemia. Although imaging features of EMC are nonspecific, its chondroid-type calcifications and foci of low signal intensity within enhancing lobules may reflect its dual histopathologic morphologic characteristics of differentiated cartilage islands interspersed within vascular undifferentiated mesenchyme. PMID- 8430194 TI - Synovial plicae and chondromalacia patellae: correlation of results of CT arthrography with results of arthroscopy. AB - The purpose of this study was to compare computed tomographic (CT) arthrography with arthroscopy in the evaluation of patients with anterior knee pain. The authors studied 23 patients with patellar plicae at CT arthrography. Twenty of 21 patients presumed to have medial plicae at arthroscopy had similar findings at CT arthrography (sensitivity, 95%; specificity, 100%). However, 10 of 12 suprapatellar plicae and 11 of 11 lateral plicae seen at CT arthrography were not seen at arthroscopy. All 11 patients with a diagnosis of no or mild chondromalacia patellae at arthroscopy had similar CT arthrographic findings, whereas seven of 11 patients presumed to have moderate or severe chondromalacia patellae at arthroscopy had matching CT arthrographic results (sensitivity, 100%; specificity, 73%). CT arthrography was much more accurate in classifying plicae as thick (66%) as opposed to classifying plicae as thin (17%). The authors conclude that CT arthrography is an accurate and specific modality for distinguishing patients with anterior knee pain due to plicae from those with anterior knee pain due to differing causes. PMID- 8430195 TI - Human immunodeficiency virus infection: musculoskeletal manifestations. AB - The authors retrospectively reviewed the charts, radiographs, and other accompanying imaging studies of 45 patients with musculoskeletal abnormalities associated with human immunodeficiency virus (HIV) infection. These included 19 patients with osseous infection, including eight with osteomyelitis, seven with bacillary angiomatosis (six of whom were described in a previous report), and four with septic arthritis; 10 with bacterial myositis (six of whom were described in a previous report); seven with non-Hodgkin lymphoma; five with hypointense marrow signal intensity at magnetic resonance imaging; two with Kaposi sarcoma; one with polymyositis; and one with psoriasis. The musculoskeletal system can be affected by a variety of abnormalities in association with HIV infection. Knowledge of their existence and characteristic appearance is valuable to radiologists for diagnosis and to clinicians for detection and appropriate treatment. PMID- 8430196 TI - Analysis of echotexture of tendons with US. AB - To evaluate changes in echotexture of normal tendons at different frequencies and establish an anatomic correlation for fibrillar echoes, normal calcaneal tendons were examined in vitro at 7.5, 10, 13, and 15 MHz in calves (n = 8) and sheep (n = 6) and in vivo in humans (n = 8). Histologic correlation was obtained in vitro with 22-gauge needles that marked the position of echogenic fibrils under ultrasound (US) guidance. The human study group consisted of 25 patients with a clinical diagnosis of calcaneal tendon disease and 15 patients who underwent surgery for rupture of the Achilles tendon. At all four frequencies, normal tendons showed an internal network of fine parallel and linear fibrillar echoes that became more numerous and thinner as US frequency increased. These echoes were caused by specular reflections at the interface between collagen bundles and endotendineum septa. In patients, tendons showed a variety of basic changes in fibrillar pattern: increased fibrillar thickness (33 patients), interruption (17 patients), fragmentation (12 patients), and disappearance of echotexture (15 patients). It is concluded that US holds promise in detection of minimal changes in tendinous structure. PMID- 8430197 TI - Periarticular bone sites associated with traumatic injury: false-positive findings with In-111-labeled white blood cell and Tc-99m MDP scintigraphy. AB - The authors evaluate the reliability of combined indium-111-labeled white blood cell (WBC) and technetium-99m methylene diphosphonate (MDP) bone scan interpretations at sites of suspected periarticular osteomyelitis with radiographic evidence of adjacent traumatic arthropathy. A review of all orthopedic patients who underwent In-111 WBC-Tc-99m MDP scintigraphy over a 7 year period revealed a subset of 32 such cases that also included results of bone biopsy cultures. Twenty-eight patients had a history of traumatic intraarticular injury, and four had periarticular fracture malunion or nonunion. Compared with intraoperative culture results, blinded In-111 WBC-Tc-99m MDP scan interpretations included four true-positive, 17 true-negative, and 10 false positive results, and one false-negative result. The predictive values for positive and negative scans were 28% and 94%, respectively. A high prevalence of false-positive In-111 WBC-Tc-99m MDP scans may occur at periarticular sites of patients with associated traumatic arthropathy. This reduces the specificity of this technique for osteomyelitis, making culture confirmation of positive scans necessary. A negative scan is highly predictive of negative culture results at these sites. PMID- 8430198 TI - Renal function in patients at risk of contrast material-induced acute renal failure: noninvasive, real-time monitoring. AB - Real-time changes in renal function were studied in a group of 20 patients at risk of contrast-material-induced acute renal failure during different angiographic procedures. Renal function was evaluated with an ambulatory renal monitor (ARM) after a single injection of the glomerular filtration agent technetium-99m diethylenetriaminepentaacetic acid (DTPA). The ARM was used to continuously monitor the clearance of Tc-99m DTPA activity from the extracellular space in the arm of the patient during angiography. A one-compartment model was used to calculate on-line the rate constant for clearance of Tc-99m DTPA from the extracellular space. Changes in the rate constant were compared with changes in plasma creatinine level measured 1-4 days after angiography. The results showed that the ARM measured rapid changes in renal function during angiography with a resolution time of 5-10 minutes in patients with normal to moderately decreased renal function and 15-20 minutes in patients with severe renal dysfunction. The sensitivity of this technique was superior to that of plasma creatinine level analysis. PMID- 8430199 TI - Triple-dose versus standard-dose gadopentetate dimeglumine: a randomized study in 199 patients. AB - To investigate the safety, patient tolerance, and efficacy with 0.3 mmol/kg gadopentetate dimeglumine in magnetic resonance (MR) imaging of the central nervous system (CNS), a phase 3 trial was conducted in 199 patients with suspected CNS lesions. Patients received either 0.1 or 0.3 mmol/kg gadopentate dimeglumine (injection time, 15 seconds and 45 seconds, respectively). T1- and T2 weighted spin-echo sequences were performed at either 0.5 T or 1.5 T. In 80 patients with enhancing brain lesions, contrast-to-noise ratios (C/Ns) were calculated, and lesion-to-brain contrast was evaluated visually. Six patients (6%) in each dose group reported adverse events. Eight adverse events occurred with 0.1 mmol/kg and seven with 0.3 mmol/kg. Vital signs and laboratory values did not change significantly. C/N (P < .05) and visual assessment ratings were higher with 0.3 mmol/kg than with 0.1 mmol/kg. According to these preliminary results, 0.3 mmol/kg gadopentetate dimeglumine is safe and well tolerated when administered at approximately 1 mL/sec. PMID- 8430200 TI - Hemodynamics of failing dialysis grafts. AB - Pressures were measured in the graft and the central vein during 104 consecutive angiographic examinations of failing hemodialysis grafts. Stenosis severity greater than 40% led to a statistically significant rise in graft pressure. In grafts in which all stenoses were of 40% or less severity, the systolic pressures in the venous and arterial limbs of the grafts were 31% +/- 16 and 45% +/- 17, respectively, of systemic systolic pressure. In grafts in which the highest grade of stenosis was greater than 40%, pressures in the venous and arterial limbs of the grafts were 53% +/- 25 and 75% +/- 24, respectively, of systemic systolic pressure. Graft thrombosis tended to occur at a higher degree of lumen reduction (but at similar pressures) with central vein stenoses compared with venous anastomotic stenoses. Dialysis graft pressures can help determine the hemodynamic importance of stenoses and the need for intervention. Percutaneous intervention should achieve arterial and venous limb pressures of less than 50% and 33% of systemic pressure, respectively. PMID- 8430201 TI - Effects of intrathrombic administration of prostaglandin E1 during pulse-spray thrombolysis with tissue-type plasminogen activator in experimental thrombosis. AB - The effects of intrathrombic and intravenous injection of prostaglandin E1 (PGE1) during pulse-spray thrombolysis were studied in a rabbit model. Thrombi were produced in the inferior vena cava of 46 rabbits by means of vessel wall injury and placement of steel coils. At 2 days, pulse-spray thrombolysis was performed by using a catheter with multiple side holes spanning the clot. A control group received injections of saline. In all other rabbits, 3 mg of tissue-type plasminogen activator (tPA) was injected into the thrombus over 1 hour, and 500 U of heparin was intravenously administered. PGE1 was administered either intravenously (50 micrograms) or directly into the thrombus (10, 25, or 50 micrograms) in four groups. After treatment, the rabbits were killed and the residual clot was weighed. The relationship between clot length and volume measured on angiograms and clot weight was assessed in 13 additional untreated rabbits. Linear regression analysis was used to estimate the initial clot weight on the basis of measured clot length in treated animals. The extent of clot lysis was significantly greater with injection of tPA and 10-, 25-, or 50-micrograms PGE1 directly into the thrombous than with administration of tPA alone. PMID- 8430202 TI - Effect of RR interval variation on image quality in gated, two-dimensional, Fourier MR imaging. AB - The effect of uneven repetition time (TR) intervals on artifact production during gated two-dimensional Fourier magnetic resonance image acquisition was investigated and a means of compensation presented. In standard spin-echo imaging, incomplete longitudinal recovery combines with normal respiratory heart rate variations to modulate signal intensity and create image copies and smearing along the phase direction. Modulation amplitude, frequency, and pattern, as well as machine parameters including echo time, TR, field of view, and image matrix size, have been found to have a substantial impact on artifact position and intensity. In a group of 10 healthy volunteers, respiratory-induced heart rate modulation produced an average of 17.2% +/- 4.9 variation in the raw data signal, with a mean frequency of 0.36 Hz +/- 0.06. The results of this study indicate that image artifacts caused by heart rate variations within the imaging period are predictable and may be reduced by correcting signal intensity for each phase view before two-dimensional Fourier transformation. PMID- 8430203 TI - Advantages of CT and beam's eye view display to confirm the accuracy of pelvic lymph node irradiation in carcinoma of the prostate. AB - Possible advantages of computed tomography (CT) and beam's eye view (BEV) display in the design of pelvic irradiation fields were studied in 20 consecutive patients with localized adenocarcinoma of the prostate. Pelvic fields were designed with standard four-field techniques. Then, CT and BEV display were done to define the reduced prostate tumor volumes. With treatment-planning CT, the location of the pelvic vascular structures (internal and external iliac artery and vein) was outlined. These were used as an approximation of the location of the lymph nodes. A BEV display of these lymph node volumes was then compared with the pelvic fields designed without CT and BEV display. Nineteen of the 20 patients had part of the CT-defined lymph node volume (a portion of the internal iliac lymph node volume) excluded from the original field design. Thirteen patients also had part of the external iliac nodal volume excluded. Dose-volume histograms showed that up to 30% of the lymph node volume received only 56% of the prescribed dose. PMID- 8430204 TI - Use of a Colapinto needle in US-guided transvaginal drainage of pelvic abscesses. AB - The ultrasound-guided transvaginal approach offers a direct, nonsurgical means for drainage of deep pelvic abscesses. It is often difficult, however, to place drainage catheters through the vaginal wall by using routine Seldinger technique. The authors describe four recent drainage procedures facilitated through use of a Colapinto needle as a dilator. In two cases, the Colapinto needle also served as a stiffening cannula for the passage of a fascial dilator. PMID- 8430205 TI - Exchange of an occluded nephroureteral stent by using a snare. AB - Percutaneous retrograde nephroureteral stent exchange may be complicated by catheter occlusion with calcareous debris. When a guide wire cannot be passed through the catheter, a snare may be advanced over the stent into the renal pelvis, permitting withdrawal of the occluded stent while maintaining access for placement of a new stent. This procedure was performed successfully in a 73-year old man. PMID- 8430206 TI - Evaluation of truncated readout-echo MR images obtained at 0.35 T. AB - It is possible to acquire a truncated echo, in which part of the information at the beginning or end of the echo is missing, and to restore the missing information by conjugation. This process was shown by means of edge spread function in phantoms and brain images in volunteers to introduce little, if any, degradation of image quality and can be used to improve sequence efficiency or to shorten echo time. PMID- 8430207 TI - Malignant tumors invading chest wall: treatment with CT-directed implantation of radioactive seeds. AB - Computed tomography (CT) was used to direct permanent implantation of radioactive iodine-125 seeds in two patients with unresectable lung cancer and in one with recurrent breast cancer invading the chest wall. An average of 60 seeds were implanted, with a mean total radioactivity of 35.6 mCi (1,317 MBq). Tumor coverage was adequate and pain relief was good in all patients. One patient had histologically documented complete response and another had CT-documented partial response. PMID- 8430208 TI - Dynamic, gadolinium-enhanced MR imaging to monitor tumor response to chemotherapy. PMID- 8430209 TI - Doppler velocimetry in the evaluation of ovarian neoplasms. PMID- 8430210 TI - Pelvic prolapse. PMID- 8430211 TI - The study of brain slices from hibernating mammals in vitro and some approaches to the analysis of hibernation problems in vivo. PMID- 8430212 TI - Glucose and ketone body utilization by the brain of neonatal rats. PMID- 8430213 TI - The ontogenesis of the forebrain commissures and the determination of brain asymmetries. AB - We have reviewed the organization and development of the interhemispheric projections through the forebrain commissures, especially those of the CC, in connection with the development of brain asymmetries. Analyzing the available data, we conclude that the developing CC plays an important role in the ontogenesis of brain asymmetries. We have extended a previous hypothesis that the rodent CC may exert a stabilizing effect over the unstable populational asymmetries of cortical size and shape, and that it participates in the developmental stabilization of lateralized motor behaviors. PMID- 8430214 TI - Newer aspects of the reversible inhibitor of MAO-A and serotonin reuptake, brofaromine. AB - 1. The reasons for developing second-generation MAOI are outlined. The expected advantage of reversibility as a safety valve with respect to tyramine potentiation is discussed. 2. Earlier data from in vitro and some ex vivo experiments had suggested an irreversible interaction of brofaromine with MAO-A, whereas the short duration of action, the absence of cumulation of effect and the displaceability by endogenously released substrates indicated reversibility. This apparent conflict could be solved by the demonstration that brofaromine behaves as a tight-binding reversible inhibitor. 3. In in vivo binding experiments with [3H]brofaromine given i.v., clorgyline, brofaromine and moclobemide were shown to dose-dependently displace the radioligand from MAO-A in the rat brain when administered after it. In corresponding experiments in the rat intestine in which the radioligand was administered p.o., similar results were obtained. Moreover, tyramine given orally in pressor doses after the radioligand also displaced it, confirming the idea that reversibility could act as a safety valve. 4. The evidence from animal and human experiments is presented that brofaromine is safer than classical MAO inhibitors with respect to tyramine potentiation. 5. Based on computer simulations, it is suggested that reduced liability of the new MAO reversible inhibitors to cause tyramine potentiation may potentially be linked to a reduced therapeutic efficacy. 6. The evidence is discussed that 5-HT uptake inhibition by brofaromine is relevant in its therapeutic effect in humans and may synergize with MAO-A inhibition, thus enhancing the impact of the latter on serotonergic transmission. PMID- 8430215 TI - Intermediary metabolism disturbance in AD/SDAT and its relation to molecular events. AB - 1. Early-onset dementia of Alzheimer type (EODAT; AD) and late-onset dementia of Alzheimer type (LODAT; SDAT) are heterogenous in origin. 2. A common superordinate pathobiochemical principle in the etiopathogenesis of both types of dementia is neuronal energy failure with subsequent abnormalities in cellular Ca2+ homeostasis and glucose-related amino acid metabolism. 3. These metabolic abnormalities are assumed to occur first at axodendritic terminals of the acetylcholinergic-glutamatergic circuit and to cause morphological damage at synaptic sites. 4. Metabolic stress and structural damage at synaptic sites may induce enhanced formation of APP and its cleavage product amyloid. 5. Energy metabolism related abnormalities along with functional and structural changes at synaptic sites of the acetylcholinergic-glutamatergic circuit may precede the formation of amyloid in DAT brain. PMID- 8430216 TI - Effects of zopiclone, flunitrazepam, triazolam and levomepromazine on the transient change in sleep-wake schedule: polygraphic study, and the evaluation of sleep and daytime condition. AB - 1. The effects of zopiclone 10mg (ZP), flunitrazepam 1mg (FN), triazolam 0.25mg (TZ) and levomepromazine 5mg (LP) on two models of sleep-wake schedule change-6 hours advanced shift: (A-shift), and 6 hours delayed: (D-shift)--were investigated in 6 healthy volunteers using polysomnography. 2. In A-shift with placebo, TST, SEI, %SR and REM/NREM decreased. ZP, FN and TZ shortened SL. All drugs increased TST and SEI. TZ and LP increased %SR and REM/NREM. 3. In D-shift with placebo, TST decreased, SWSL was prolonged, %S3+4 decreased, and %SR and REM/NREM increased. All drugs increased TST. ZP and LP shortened SWSL. All drugs increased %SWS. ZP, FN and TZ decreased %SR. ZP and FN decreased REM/NREM. 4. Daytime mental and physical conditions were worse than usual on more than half of the days in A- and D-shift. LP and FN caused some inadequate conditions on the following days. Significantly higher REM/NREM was observed in the nights before the days with worse mental conditions in D-shift, and lower REM/NREM in the nights before the days with worse physical conditions in A-shift. 5. It is concluded that TZ and ZP are superior to the others for A-shift and D-shift, respectively. PMID- 8430217 TI - Significantly increased expression of T-cell activation markers (interleukin-2 and HLA-DR) in depression: further evidence for an inflammatory process during that illness. AB - 1. Recently, the authors have reported that severe depression may be accompanied by a systemic immune activation with an increase in the number of T cells expressing activation receptors. 2. The present large-scale study examines specific T (CD2+HLADR+ and CD7+CD25+) and B (CD7-CD25+) cell activation markers in depressed inpatients and normal volunteers together with the number of leukocytes and monocytes. 3. The authors have established that depression is characterized by a significantly increased expression of T cell activation receptors (CD7+CD25+) and by the appearance of previously unexpressed T cell surface markers (CD2+HLADR+). There was a significant and positive correlation between the number of CD7+CD25+ cells and monocytes, with the expression of the HLADR and CD25 T cell activation markers being significantly and positively correlated. Up to 64% of all depressed subjects exhibit an increased expression of these activation markers with a specificity of 91%. 4. The normal control group and the depressive sample constitute two discrete classes (i.e., qualitatively distinct groups) with respect to the expression of these activation markers and leukocytosis. 5. It is concluded that our results are compatible with the presence of T-cell activation in a considerable number of depressed patients. PMID- 8430218 TI - Relation of plasma and red blood cells reduced haloperidol concentrations to haloperidol reductase activity assayed in red blood cells in psychiatric population. AB - 1. Haloperidol (HAL) reductase activity in red blood cells (RBC) was determined by a newly developed assay method in 120 blood samples from 75 Japanese psychiatric patients receiving HAL. 2. Plasma concentrations of HAL and reduced haloperidol (RHAL), a reductive metabolite of haloperidol, were also measured in these samples. 3. RBC concentrations of HAL and RHAL were measured in 62 of these samples. 4. No significant correlations were found between HAL reductase activity in RBC vs plasma or RBC RHAL/HAL ratios, which may represent activity of the enzyme metabolizing HAL into RHAL. 5. RHAL concentrations were three times higher in RBC than in plasma, though HAL concentrations were at the same level in both tissues. This may reflect accumulation of RHAL in RBC. PMID- 8430219 TI - Functional effects of glucocorticoid exposure during fetal life. AB - 1. Pregnant rats were exposed to hydrocortisone in a dose of 10 mg/kg on days 9 11 or days 13-15 of gestation. The offsprings born to these mothers were observed for their behavioral development and the response to kainic acid during the infantile period. The response to kainic acid was assessed by the frequency of wet-dog shakes behavior and limbic seizures. 2. The growth rate in the infantile offspring of the 13-15dHc-F1 group showed a slight but significant decrease. 3. All the 13-15dHc-F1 rats exhibited the rearing activity in an open-field at 17 days of age, earlier than in the controls. 4. The ambulatory activity in the 9 11dHc-F1 rats showed a significant decrease at 15 and 17 days of age, whereas no change was shown in the 9-11dHc-F1 rats. 5. The frequency of the wet-dog shakes during the 60 minutes observation after the s.c. injection of 9 mg/kg kainic acid was significantly low in both the 9-11dHc-F1 and 13-15dHc-F1 groups as compared with that in the controls when tested at 25 days of age. The decrease in the response to kainic acid was slightly greater in the 9-11dHC-F1 rats. 6. The frequency of seizures with forelimb clonus and rearing during the 60 minutes observation in the 13-15dHc-F1 was less than that in the controls, whereas no significant difference in the frequency of seizures between the 9-11dHc-F1 and paired control groups was noted. 7. The second generation rats raised from the 9 11dHc-F1 rats by brother-sister mating showed a decrease in the frequency in the kainic acid-induced wet-dog shakes as shown in the F1 offspring. No change in the response to kainic acid was shown in the 13-15dHc-F2 rats. PMID- 8430220 TI - The effects of valproate in brain monoamines of juvenile rats after stress. AB - 1. Clinical data suggest that valproate (VPA) may be useful in prophylaxis of affective disorders, which show disturbances of the serotoninergic system. On the other hand, chronic stress has an adverse effect on affective disorders, those with disturbances of the serotonergic system, especially. 2. In order to study the effects of VPA on brain monoamines and acute stress, 200 mgr VPA/Kgr was administered intraperitoneal (ip) to juvenile male rats; the control group was treated with NaCL 0.9% ip. After 30 min, all animals were evoked on predictable neurogenic or systemic stress (30 min foot shock, or 15 min ether stress, respectively), and 48 hours later, VPA or NaCL were administered ip again; 30 min afterwards, the rats were decapitated. Rats without stress were also sacrificed 30 min after VPA or NaCL administration. 3. Measurements of brain monoamines noradrenaline (NA), dopamine (DA), 5-hydroxytryptamine (5-HT), and their metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA), were done in Frontal Cortex (FC), Hypothalamus (HY) and Striatum (S), by High Performance Liquid Chromatography (HPLC). 4. Compared with the control stress group the level of 5-HIAA in the FC was significantly increased (P < 0.01) in VPA stress rats; in the HY and in S the increase of 5-HIAA was not significant. No remarkable differences were observed in NA, DA, 5-HT and DOPAC concentrations, in any of the brain regions. No changes in brain monoamine levels were found in non stress rats, either. 5. The augmentation of 5-HIAA level after VPA administration and after stress, in correlation with the decrease of 5-HIAA that is observed in depression, support the hypothesis that VPA may be effective in affective disorders by influencing the serotoninergic system. PMID- 8430221 TI - Skin conductance reaction (SCR)-habituation test, an elementary model of anxiety as a tool to assess anxiogenic activities of some drugs. AB - 1. The SCR-habituation test, used as a tool to detect an anxiolytic activity, is applied to the study of several anxiogenic drugs. This test is based on the measure of the SCR-extinction time (i.e. habituation) got by the way of an iterative photostimulus. 2. 8 anxiogenic compounds are tested. They delay significantly and quantitatively the SCR habituation. 3. Specificity, sensitivity and quantifying values of the test are considered in order to propose the use of the SCR-habituation test in the investigation of putative anxiogenic compounds. PMID- 8430222 TI - Expression of a common cellular phospholipase A2 by human intrauterine tissues. AB - A subclone of the three prime (3') end of a human extrapancreatic phospholipase cDNA has been generated using the polymerase chain reaction. This has been used to detect mRNA transcripts in RNA extracted from human amnion, chorion-decidua, placenta and myometrium. The carboxyl-terminus of phospholipases A2 is poorly conserved but hybridisation remained stable under conditions of high stringency. This supports the hypothesis that these tissues express a common cellular phospholipase A2 which is identical to that expressed in platelets. PMID- 8430223 TI - Primary colonic epithelial cell culture of the rabbit producing prostaglandins. AB - We have established primary colonic epithelial cell culture from adult rabbits and examined effects of anti-inflammatory drugs on prostaglandin (PG) E2 production. Colonic epithelium of adult rabbits was scraped and minced into small pieces. They were incubated for isolation in Hanks' balanced salt solution with 0.35% collagenase and Earle's solution with 1 mM EDTA. Isolated cells were cultured in Coon's modified Ham's F-12 medium with 10% fetal bovine serum and antibiotics on collagen coated cell wells. The medium was refed twice a week. The production of PGs was assessed by high pressure liquid chromatography (HPLC). PGE2 and PGF2 alpha were measured by radioimmunoassay. Within 24 hours after inoculation, the cell clumps attached to the surface of the wells and cells began to spread out and grow. Monolayer cultures became confluent in 4 days. Phase contrast microscopy showed that these cells consisted of a homogeneous population of epithelial cells with large oval nuclei, polyhedral shape, and organized sheet like growth pattern. HPLC profile showed synthesis of 6-keto-PGF1 alpha, thromboxane B2, PGF2 alpha, PGE2, and PGD2 by cultured cells. Quantitatively, 117 +/- 7 ng/mg-protein/hour PGE2 and 7.4 +/- 0.7 ng/mg-protein/hour PGF2 alpha were produced. While hydrocortisone (10(-4) - 10(-2) M) did not show a significant effect on PGE2 production, indomethacin (10(-8) - 10(-6) M), and 5-aminosalicylic acid (2X10(-4)-5X10(-3) M) inhibited PGE2 production. We have established relatively convenient procedure for primary culture of colonic epithelial cells from adult rabbits. Different actions of anti-inflammatory drugs on PGE2 synthesis suggest that these cultured cells might be a good tool for the various cellular functional studies of normal colonic epithelial cells. PMID- 8430224 TI - Acetylsalicylic acid inhibits anticardiolipin antibody-induced platelet activating factor (PAF) synthesis. AB - Enhanced endothelial cell PAF synthesis has been identified as a consequence of anticardiolipin antibody (ACA)-positive serum exposure. We proposed this observation as a contributing factor to thrombogenesis in women with the antiphospholipid syndrome. Since acetylsalicylic acid (ASA) is an accepted therapeutic alternative in these patients, we sought to determine if ASA would attenuate endothelial cell PAF production resulting from ACA exposure. Using primary, confluent monolayers of umbilical vein endothelial cells, experiments were performed to evaluate PAF synthesis after incubation with antibody-positive serum and ASA. Total PAF and its radyl-derivatives (1-alkyl- and 1-acyl-PAF) were quantified by tritiated acetate incorporation, phospholipid extraction, thin layer chromatography and scintillation spectroscopy. ASA consistently decreased ACA-induced PAF synthesis (No ASA, 9573 +/- 443 vs 1mmol/L ASA, 4829 +/- 838 dpm/ml; p = 0.016) and the observed reduction was dose-dependent over a range of ASA concentrations (0.1, 1, 10 and 100 mmol/L; ANOVA, p = .00015). Reduced PAF synthesis was also observed in cultures exposed to ASA and incubated with antibody-negative serum. These observations suggest that in ACA-positive women, the antithrombotic effects of ASA may relate in part, to reduced endothelial cell PAF synthesis. PMID- 8430225 TI - Effects of epidermal growth factor on serum zinc and plasma prostaglandin E2 levels of mice with pressure sores. AB - This work was undertaken to study the effects of systemic application of EGF on the plasma PGE2 and serum zinc levels of mice with pressure sores. The pressure sores were made on the hind limb of the mice by bilateral sciatic nerve neurotomy. Mice were injected IP, 1 microgram EGF daily for ten days, beginning on the 21st day after operation. EGF administration increased the serum zinc and plasma PGE2 levels compared with controls. These results indicate that EGF can be effective on wound healing by elevating the serum zinc and plasma PGE2 concentrations together with other physiological roles in cellular events. PMID- 8430226 TI - PGF2 alpha, PGE2, progesterone, and estradiol-17 beta, secretion by the corpus luteum of the oviparous lizard, Podarcis sicula sicula. In vitro studies. AB - The release in vitro of prostaglandin F2 alpha (PGF2 alpha), prostaglandin E2 (PGE2), progesterone, androgens and estradiol-17 beta by the corpora lutea (CL) of the oviparous lizard, Podarcis s. sicula, was studied. In addition, the in vitro effects of PGF2 alpha and PGE2 on sex steroid release by CL were evaluated. Corpora lutea were divided into four types, according to their different developmental stage: CL1 (unshelled eggs in the oviduct); CL2 (shelled eggs in the oviduct); CL3 (eggs laid 6 h previously); CL4 (eggs laid 48 h previously) and were placed into culture. PGF2 alpha secretion was highest in CL4 incubated samples and lowest in CL2 and PGE2 was highest in CL1 and CL2. Progesterone secretion was highest in CL2 and lowest in CL4; androgens were not detectable and estradiol-17 beta secretion was highest in CL2. PGF2 alpha decreased progesterone secreted by CL1, CL2 and CL3, while it did not modify release of androgens and estradiol-17 beta. PGE2 did not affect sex steroid release. These data suggest a role of PGF2 alpha in inducing luteolysis, while PGE2 could be implied in the maintenance of CL. A role of progesterone during gestation of Podarcis s. sicula was also confirmed. PMID- 8430227 TI - PGE2 attenuates PGF2 alpha-induced increases in free intracellular calcium in ovine large luteal cells. AB - When ovine large luteal cells are placed in culture and exposed to PGF2 alpha, there is a rapid and sustained increase in the concentration of free intracellular calcium which is believed to play a major role in the luteolytic and cytotoxic effects of PGF2 alpha. Since administration of exogenous PGE2 can prevent spontaneous and PGF2 alpha-induced luteolysis in vivo, and the cytotoxic effects of PGF2 alpha on large luteal cells in vitro, the objective of this study was to determine if one mechanism by which PGE2 acts is to attenuate increases in free intracellular calcium induced by PGF2 alpha. At concentrations of 10 nM or greater, PGF2 alpha caused a significant and sustained increase in free intracellular calcium in large luteal cells. Similarly, PGE2 also induced increases in free intracellular calcium but required doses 20-fold greater than PGF2 alpha. When PGE2 (1, 10 or 100 nM) was incubated with PGF2 alpha (100 nM) increases in free intracellular calcium induced by PGF2 alpha were attenuated (P < 0.05) when measured 5 min, but not at 30 min, after initiation of treatment. The observed decrease in the concentration of free intracellular calcium at 5 min in response to PGF2 alpha was the result of fewer cells responding to PGF2 alpha. In addition, the concentrations of free intracellular calcium attained in the cells that did respond was reduced 25% compared to cells treated with PGF2 alpha alone. Thus, part of the luteal protective actions of PGE2 appears to involve an inhibition of the early (5 min) increase in free intracellular calcium induced by PGF2 alpha. PMID- 8430228 TI - Microdialysis measurements of PGD2, TXB2 and 6-KETO-PGF1 alpha in rat CA1 hippocampus during transient cerebral ischemia. AB - We measured hippocampal CA1 concentrations of PGD2, TxB2 and 6-keto-PGF1 alpha in 18 anesthetized rats with stereotaxically implanted microdialysis probes before, during and after 20 min of global cerebral ischemia. The insertion of the microdialysis probe did not appear to cause a continuous major disturbance of arachidonic acid (AA) metabolism because stable eicosanoid concentrations were obtained prior to ischemia. During reperfusion all three eicosanoids increased significantly reaching a peak after 30-60 min and then gradually declined to baseline levels over the next 2-3 h. The ratio of average peak concentrations for PGD2, TxB2 and 6-keto-PGF1 alpha were approximately 80:2:1, respectively. The results extend previous work by demonstrating the time course of eicosanoid release in a distinct brain region and confirm the role of PGD2 as the major PG metabolite in brain. We conclude that future studies employing microdialysis may be able to provide a more detailed understanding of the role of AA metabolites in ischemic brain. PMID- 8430229 TI - [Superstructure or foundation? On metapsychology and the metapsychology debate]. AB - In the sixties and seventies the value (or otherwise) of Freud's metapsychology was the subject of animated discussion. Today the subject seems to have lost its power to excite controversy. The author reconstructs the criticism of metapsychology and shows that the problems posed by the "witch" metapsychology can by no means be regarded as shelved. If we locate metapsychology in the boundary area between psychic and somatic and regard it as an attempt to conceptualise the unity of body and mind/soul, then the author feels it necessary to hold on to metapsychology as a crucial source of psychoanalytic knowledge. Further, she calls for a research programme with a historical perspective to establish an archeology of the argumentative structures underlying metapsychological discourse since Freud. PMID- 8430230 TI - [Organization of psychoanalytic institutions: psychoanalysis can't function without, psychoanalysis can't function with it? Or indeed?]. AB - Recently there has been much heated debate on the question of the effects of the institutionalisation of psychoanalysis on analysis itself and the significance and consequences of the decision of psychoanalysts to organise themselves in professional organisations. The author identifies two errors analysts are prone to make when dealing with institutions/organisations. One is to confuse psychodynamics with the dynamics of institutions, the other to look upon an organised professional grouping as a species of family to which one can turn for assistance and encouragement. To rectify this misunderstanding, the author calls for a strict distinction between two very different levels of reality. "Non trivial machines" such as psychoanalytic institutes, characterised by a high degree of complexity, obey a logic of their own which cannot be grasped in psychodynamic terms or in analogy to family relations. Buchinger sees it as essential to learn to think and act in terms of structures as opposed to persons and relations. Such a learning process can only be successful if specifically psychoanalytic self-reflection is complemented by an appropriate form of organisational self-reflection. PMID- 8430231 TI - [The professional status of analytic and adolescent psychotherapists in the "FOGS Study]. AB - Analytic psychotherapists for children and young people are the only group of psychoanalytic methods among "laypersons" has become reality. These therapists are integrated into the overall German health care system. The author discusses the scope of their activities as reflected by the "FOGS Study", underlining the medico-centric profile displayed by this professional group and the attendant duality characteristic of it. On the one hand, these therapists are incorporated organisationally into the professional associations of the psychoanalysts, but in their activities and trainee work they are largely autonomous. It is to be hoped that the planned "Psychotherapist Law" will provide this group with a status in which their legal standing and their specific therapeutic identity will be more closely matched. PMID- 8430232 TI - [Delinquency as an expression of narcissism and borderline disorder. Institutional and individual therapeutic elements]. AB - The author reports on his therapeutic activity with delinquent youngsters in a Swiss penal institution where the principle of pedagogical "correction" has been eschewed in favour of that of a therapeutic community. He traces the success of this institution largely to the fact that the formation of psychoanalytically oriented, theme-centered groups acts as a factor militating against splitting processes among the therapists themselves. In the author's view, delinquency derives from narcissistic and borderline disorders, discussed here with reference to Kohut and Kernberg. PMID- 8430233 TI - [The peripheral analgesic action of the exogenous nitric oxide donor: nitroglycerin. A placebo-controlled study of the transdermal action of nitroglycerin on pain sensitivity in the forearm]. AB - Nitric oxide is a biological mediator. In nervous system it acts like neurotransmitter and also modulate acute inflammation. In the peripheral nervous system it blocks the nociceptive stimulus through an increase in postsynaptic neurone GMPc level. Nitro-vasodilator drugs like nitroglycerin are metabolised in the cell given rise to short lived intermediates, which liberating nitric oxide that activate the guanylate cyclase enzyme, increasing the GMPc in smooth muscle cell. This study show that nitroglycerin produces an analgesic action. The pain sensitivity to pinprick test in forearm with nitroglycerin has shown a decrease in a significative manner against placebo. We speculate that nitroglycerin could have a similar action as endogenous nitric oxide in nervous system. PMID- 8430234 TI - [Chest pain of esophageal origin in patients with a normal coronary angiogram and ergonovine-induced coronary spasm]. AB - In order to differentiate the cardiac or oesophageal origin of chest pain, 55 patients with chest pain, normal coronary arteriogram and normal left ventricular function, were studied. Patients were evaluated with ergonovine test to induce coronary artery spasm and oesophageal function study (including basal manometry in all cases, ClH acid instillation in 53, manometry during ClH instillation in 32 and edrophonium test in 9). There was coronary artery spasm following ergonovine test in 8 patients (group 1) and negative results in 47 (group 2). There was oesophageal disfunction in 50% patients in group 1 and in 62% patients in group 2 (p = NS). The incidence of motor disorders or chest pain following acid instillation was not significatively different in both groups. Nevertheless, in group 1 a tendency to a greater incidence of oesophageal spasm was observed while in group 2 unspecified disorders were more frequent. Thus, in patients with chest pain and normal coronary arteriogram, we always must discard coronary artery spasm and oesophageal disfunction, because, due to a probably common cause, association between both disorders is frequent. PMID- 8430235 TI - [Uniform requirements for manuscripts submitted to biomedical journals. The International Committee of Medical Journal Editors]. PMID- 8430236 TI - [Transesophageal echocardiography in the assessment of coronary artery anatomy and blood flow]. AB - Proximal coronary artery anatomy and coronary blood flow was evaluated by transesophageal echocardiography in 41 consecutive patients without coronary artery disease. The left main coronary artery was visualized in 37 patients (90%), the left anterior descending coronary artery in 20 (48%), the left circumflex artery in 16 (39%) and the right coronary artery in 7 (17%). The image quality was optimal in 58% of patients in left main coronary artery, 29% in left anterior descending coronary artery, 24% in left circumflex artery and 5% in right coronary artery. Transesophageal Pulsed Doppler echocardiography evaluation of blood flow velocity in the left coronary artery was obtained in 37% of patients. Velocity flow pattern was biphasic with greater diastolic component and smaller systolic one (average peak flow velocity of 47.5 +/- 16.6 cm/seg during diastole and 23.1 +/- 9.5 cm/seg during systole). Coronary blood flow was detected by transesophageal color Doppler echocardiography in 51% of patients in left main coronary artery, 24% in left anterior descending coronary artery, 19% in left circumflex artery and 2% in right coronary artery. We conclude that transesophageal echocardiography provides better and more accurate visualization of proximal coronary arteries than transthoracic echocardiography, but technical limitations still remain to detect and evaluate coronary artery anatomy and coronary blood flow in a large number of patients. PMID- 8430237 TI - [Malignant vasovagal syncope: its diagnosis and therapeutic trial based on the orthostatism test (the tilt-table test)]. AB - Seventy consecutive patients with recurrent unexplained syncope were evaluated by use of an up-right tilt-table test for 45 minutes (with or without an infusion of isoproterenol) in an attempt to reproduce symptoms. There were 42 males and 28 females with a mean age of 49 +/- 20 years (range 7-86), and with a mean symptoms duration of 35.2 +/- 16 months. All the patients underwent clinical examination which includes cardiological and neurological evaluation. Some tilt positive patients received therapy with either transdermal scopolamine, metoprolol or clonidine, the efficacy of which was evaluated by another tilt-table test. Syncope occurred in sixteen patients (22.9%), during the baseline tilt associated with hypotension (2 patients), bradycardia-asystolia (3 patients), or both (11 patients). In 11 patients with normal baseline tilt test, isoproterenol infusion was used appearing 4 new cases of abnormal response. Some patients who had positive test results, eventually became tilt-table negative by therapy (5 out of 8 patients treated with scopolamine, 5 out of 6 patients with metoprolol, and 3 out of 5 patients with clonidine). Time before syncope during tilt-test was increased with therapy, being of 45 min in normalized cases, and increasing from 11.9 +/- 16 basal to 34.4 +/- 17 min with scopolamine (p < 0.05), to 40 +/- 12 min with metoprolol (p < 0.05) and to 33.7 +/- 16.7 min with clonidine (p = NS). We conclude that upright tilt-table alone or combined with isoproterenol infusion is an useful test in the diagnosis of vasovagal syncope and in the evaluation of therapy. PMID- 8430238 TI - [Cardiac arrhythmias (I). The therapeutic options in the decade of the nineties]. PMID- 8430239 TI - [The transcatheter closure of the persistent ductus arteriosus in an adult]. AB - Transcatheter closure of a patent ductus arteriosus was successfully performed in a symptomatic 48-year-old man. Pulmonary to systemic flow ratio was of 2.8:1 and ductus arteriosus closure was carried out percutaneously, using a 17 mm. Rashkind umbrella. One month later, the patient presented a significant clinical improvement and there also was a reduction of the left ventricle dilatation, objectively assessed by two-dimensional echocardiography. PMID- 8430240 TI - [Acute myocardial infarct and coronary artery aneurysm during pregnancy]. AB - We present the first published case in Spain on a gestant woman with acute myocardial infarction. She was angiographically studied showing a coronary artery aneurysm with inner thrombus. This is also the first time that such a lesion is described in vivo in a gestant woman. PMID- 8430241 TI - [Cardiac rhabdomyosarcoma. Its echographic diagnosis]. AB - We report a case of cardiac rhabdomyosarcoma whose initial clinical features were fever and palpitations due to documented ventricular tachycardia. Sequential two dimensional echocardiographic studies pointed out the presumed diagnosis, showing intracavitary masses at multiple sites appearing within a short period of time. The postmortem examination confirmed the diagnosis of rhabdomyosarcoma. PMID- 8430242 TI - [Accidental hypothermia, rewarming by the use of extracorporeal circulation]. PMID- 8430243 TI - [Prison, drug addicts and AIDS]. PMID- 8430244 TI - [Admission to a degree course in medicine and surgery]. PMID- 8430245 TI - [Hemorrhages of the upper gastrointestinal tract and nonsteroidal anti inflammatory drugs: the results of a case-controlled surveillance]. AB - A postmarketing surveillance case-control study was set up and applied in an Italian Hospital network to quantify the risk of upper gastrointestinal bleeding (UGB) and exposure to non steroidal anti-inflammatory drugs (NSAIDs). During the period of study 441 cases of UGB and 1323 controls were recruited. The odds ratios (OR) associated with NSAIDs use were estimated for intake occurring over two different periods of time prior to hospital admission (i.e. during the preceding week and month). A strong association emerged for aspirin intake, both in the week (ORMLR = 11.2; 95% CI 7.8-16.9) and in the month (ORMLR = 6.9; 95% CI 4.6-10.2) preceding hospital admission. [MLR = Multiple logistic regression; CI = Confidence interval]. A significant increase in the risk of UGB and use of diclofenac, phenylpropionic acid derivatives, and indomethacin was also found in the two exposure periods considered, while for piroxicam a significant association was only apparent in the analysis of 1-month exposure. As expected, paracetamol and pyrazolone derivatives were not associated with UGB. This pilot experience has shown the feasibility of setting up a multicenter post-marketing surveillance programme and of establishing a network for drug monitoring within the Italian National Health Service, capable of providing a thorough evaluation of the benefit/risk profile of drugs. PMID- 8430246 TI - Hydrogen breath test and scintigraphic gastrocolic transit time in diabetics with autonomic neuropathy. AB - We assessed gastrocolic transit time in 10 diabetic patients with autonomic neuropathy and 10 healthy age-matched controls by measuring breath hydrogen rise and scintigraphic bolus progression after ingestion of an isosmotic lactulose solution containing 99m-Tc-diethylentriamine-pentaacetic acid. Mean transit time in diabetics with autonomic neuropathy was significantly slower with respect to controls, with a good correlation between the two techniques. Moreover, diabetics had a significantly shorter discharge time (defined as the period that elapses between the arrival of the meal into the cecum and the hydrogen increase in the expired air). It is concluded that the selective lipid malabsorption seen in diabetic patients could be the result of cecal-ileal reflux, as a contaminating consequence of the last ileal loop. A possible motor innervation defect of the ileo-cecal valve is postulated in these subjects. PMID- 8430248 TI - [Neoplasm of the head of the pancreas]. PMID- 8430247 TI - [Peptic ulcer and Helicobacter pylori. Comments on the authors' cases]. AB - The presence of Helicobacter was tested on a group with antral or duodenal ulcer with or without gastritis, versus a group without gastric or duodenal pathology. Furthermore an open trial was performed between omeprazole and colloidal bismuth subcitrate (CBS) on patients similarly affected by peptic disease. Although CBS did eliminate Helicobacter in more than a half of patients, what was not obtained by omeprazole, this result did not mean a better control of peptic disease: in fact the omeprazole was remarkably more active in our series on clinical and endoscopic ground, whether the Helicobacter was present or not, and further studies are required to assess the real significance of Helicobacter pylori in the above conditions. PMID- 8430250 TI - [Omeprazole prevention of the recurrence of peptic ulcer: the efficacy of treatment with 20 mg on alternate days]. AB - Omeprazole, 20 mg every second day, in prevention of duodenal ulcer relapse has been tested. 15 patients with peptic ulcer, which was endoscopically proved and then healed, underwent the treatment. No matter of ulcer original seat as well as antacid drug previously employed, recurrences were not seen after a 6 months period, while clinical symptoms and possible adverse reactions appeared to be very slight. The proposed regimen of omeprazole seems to be cost-saving and worth proving. PMID- 8430249 TI - [The effect of colloidal bismuth and its combination with amoxicillin on the elimination of Helicobacter pylori in patients with a duodenal ulcer and chronic antral gastritis]. AB - Although the aetiopathogenetic role of Helicobacter Pylori (HP) in duodenal ulcer and in chronic gastritis seems now well defined, we have not yet standardized therapeutic schedules to achieve disappearance of HP. This study was aimed at evaluating the efficacy of two different therapeutic schedules, colloidal bismuth alone [CB(1200 mg/day)], vs. association with amoxicillin [CB+A(3 g/day)] for six weeks, to clear HP from antral specimens in a cohort of dyspeptic patients. 49 consecutive patients (23 females, 26 males, mean age 47 years, range 22-69) with HP in gastric specimens, 30 suffering from chronic antral gastritis (CG) and 19 affected by duodenal ulcer (DU) were treated with CB (37 pts) or with CB+A (12 pts). DU patients were given also H2-blockers. These latter patients were all healed at the endoscopic control performed after therapy. 4 out of 13 DU patients (31%) treated with CB were found HP free. In 4 out of 6 DU patients (66%) treated with CB+A, HP was no more detectable. As for CG patients, 12 out of 24 (50%) were free from HP at control when treated with CB, while only 2 out of 6 (33%) when CB+A was administered. This study suggests that colloidal bismuth is more effective when administered associated with amoxicillin, but this concerns only DU patients. No relation between endoscopic healing of UD and HP presence was found. PMID- 8430251 TI - [The rifaximin therapy and prophylaxis of episodes of acute diverticulitis]. AB - The authors consider some nosographic aspects of diverticular disease, and refer the personal experience on the treatment of 36 patients with diverticulitis. Beside surgery, that is indicated when complications occur (perforation, abscess, fistulae, occlusion), medical therapy (diet + drugs) could reduce the symptomatology and the risk or progression of complications. The authors present data on the clinical use of rifaximin in the treatment of acute diverticulitis. PMID- 8430252 TI - Temporal arteritis in a patient with ulcerative colitis. Coincidental association or (immuno) pathogenetic link? PMID- 8430253 TI - [AIDS patients in Italian prisons: a burning issue with no easy solution]. AB - Some of the problems related to the detention of HIV infected subjects in Italian prisons are discussed, with particular reference to subjects with AIDS. The Italian Department of Penitentiary Administration reported, in 1991, the presence of 3030 HIV infected subjects, out of which 84 with AIDS. These subjects amount to 10% of prisoners annually detained in italian prisons. Recently Italian Government, in order to regulate the management of prisoners with HIV infection, has issued a decree-law (no. 431 of November 12th, 1992). This decree sanctions the unacceptability of detaining people with AIDS or any other severe immune deficiency. Nowadays it is impossible to predict the effects of this regulation. Many factors worsen the situation: structural peculiarities and overcrowding of prisons, prisons staff anxiety, lack of awareness about uselessness of obligatory prisoners' screening. The World Health Organization in 1987, the European Council in 1989 and the Italian Committee against AIDS in many occasions, during the last five years, gave indications for the management of seropositive prisoners and confirmed that individual counselling and confidentiality of information can help the prisoners to accept the HIV test: except for a few cases, the isolation of seropositive prisoners is not justified; prevention plays an essential role. PMID- 8430254 TI - [The principles of the new regulation]. PMID- 8430255 TI - [The why of a reform]. PMID- 8430256 TI - [The reform of Schedule XVIII]. PMID- 8430257 TI - Coping patterns and related characteristics in patients with IBD. AB - This article describes a study designed to examine the coping behaviors, personality, and mood characteristics of 150 nonhospitalized adults with inflammatory bowel disease (IBD). Self-report questionnaires and interviews were used to collect the data. Coping responses were measured with the Jalowiec Coping Scale. The Eysenck Personality Questionnaire (EPQ) and Beck Depression Inventory (BDI) were used to ascertain other characteristics. Data were analyzed using analysis of variance (ANOVA), Pearson's correlation coefficient (r), and descriptive statistics. IBD patients scored significantly higher on using problem oriented coping patterns than on using affective-oriented methods. The results suggest that coping patterns in IBD patients are remarkably effective in contributing to lifestyle satisfaction. It can be inferred that adaptive efforts of IBD patients can be enhanced if health professionals consider coping patterns in the assessment process and if they actively support a sense of control in these patients. PMID- 8430258 TI - Toxoplasmic encephalitis in the AIDS patient. AB - Toxoplasmic encephalitis is recognized as a major cause of opportunistic infection of the central nervous system in patients with acquired immune deficiency syndrome (AIDS). Often this protozoal infection is benign, but in the AIDS patient it can cause life-threatening complications. Usually, toxoplasmic encephalitis is a latent infection activated in the brain in some 5% to 15% of AIDS patients. Treatment with oral antibiotics is effective, but relapse is common, and there may be permanent brain tissue damage with resultant neurological deficits. This article acquaints the rehabilitation nurse with the diagnosis and acute nursing care of toxoplasmic encephalitis. Rehabilitation nurses need to know about this chronic condition because rehabilitation is key to its successful long-term care and management. PMID- 8430259 TI - Rehabilitating a loved one: a personal story. AB - Fortunately, during their professional lives, only a small percentage of nurses will face the traumatic task of helping a loved one overcome a major physical handicap. In this article, an experienced rehabilitation nurse relates her experience in helping her aged mother cope with the reality of a severe injury and amputation. PMID- 8430260 TI - A comparison of open and closed catheterization techniques in rehabilitation patients. AB - An effective method was sought to decrease the incidence of urinary tract infections acquired by hospitalized rehabilitation patients requiring catheterization. The purpose of this study was to determine whether there was a difference in the incidence of urinary tract infections that occurred following use of two types of intermittent catheterization techniques: open catheterization and closed catheterization. The incidence of infection in both the control (open catheterization) and experimental (closed catheterization) groups was analyzed. In the control group, 9 of 14 patients (5 stroke, 4 spinal cord injured [SCI]) completed all four urine tests. Two patients developed a 20,000 colony count of enterococcus on the fourth urine sample using open catheterization. In the experimental group, 11 (4 stroke, 7 SCI) out of 16 patients maintained colony counts below 100,000 organisms per ml; one patient had a 16,000 colony count using closed catheterization. Qualitative data revealed that patients had a positive response to the closed system. PMID- 8430261 TI - The spouse's positive effect on the stroke patient's recovery. AB - To determine the effect of the presence of a spouse on the stroke patient's adaptation, a retrospective chart review of stroke patients who had completed rehabilitation was conducted. The convenience sample comprised 67 stroke patients of a family practice service who received rehabilitation at Pitt County Memorial Hospital Rehabilitation Center in Greenville, NC, over a 45-month period. Roy's Adaptation Model was the theoretical framework for the inquiry. The Barthel Index measured the functioning level, or adaptation, of the patients on discharge from rehabilitation. It was hypothesized that patients who had a stroke and completed a rehabilitation program would achieve a higher level of adaptation if they had a spouse than would comparable patients without a spouse. This hypothesis was supported by the findings; the average Barthel Index score on discharge was 7.5 points higher in patients who had a spouse (Wilcoxon chi-square = 4.17, p < .05). The implications for nurses are presented. PMID- 8430262 TI - Coping with a mentally and physically impaired infant: a self-analysis. PMID- 8430263 TI - Use of a nurse extender role in the rehabilitation setting. AB - In an effort to meet the increasing demand for bedside caregivers, nurse extender programs have proliferated over the past 2 years throughout the country. Standards for their job descriptions, however, are as varied as the number of programs. At Braintree Hospital, a great deal of time and effort has been spent to develop a program that is managed and supervised by registered nurses and that supports the appropriate delegation of tasks that do not require the skill of a professional nurse. Merker and Burkhart (1990) supported the use of a nurse extender to enhance the delivery of patient care and suggested that nursing administrators need to assess the current environment, define a strategy for the use of nurse extenders, and then implement that strategy. They also noted that because all healthcare facilities differ, it is important that each facility design its own program to meet its individual needs. PMID- 8430264 TI - The initial RNF research survey: rehabilitation nursing research interests of ARN members. AB - The data from this survey provide baseline information for comparison with subsequent surveys. Respondents to this initial survey demonstrate involvement in research activities, primarily for the direct improvement of care. Findings from the data suggest that the research climate is right for collaborative efforts between rehabilitation nurses caring directly for clients and nurses with research degrees to conduct multisite research projects and to encourage enhanced research preparation, including computer literacy, for nurses at the graduate level. PMID- 8430265 TI - Review of the World Health Organization's report on disability prevention and rehabilitation. AB - The key approaches recommended by the WHO expert committee in this standard setting document are these: 1. the prevention of disability through environmental sanitation, adequate nutrition, and supportive health care; and 2. the provision of rehabilitation services within the available and accessible primary healthcare model. Programs based on these approaches have substantially improved the quality of life for many disabled people in the Third World. Familiarity with this key international position paper is an essential starting point for the nurse interested in contributing to the field of rehabilitation in underserved areas of the world, including areas of the United States. PMID- 8430266 TI - The effects of two teaching techniques on recognition and use of function words by aphasic stroke patients. PMID- 8430267 TI - The effects of two teaching techniques on recognition and use of function words by aphasic stroke patients. PMID- 8430268 TI - Deep vein thrombosis in the rehabilitation client: diagnostic tools, prevention, and treatment modalities. AB - This article details the diagnosis, prevention, and treatment of deep vein thrombosis (DVT). Specific preventive measures discussed include the use of coumadin and of intermittent pneumatic compression stockings. Pros and cons of using knee-high and thigh-high antiembolism stockings also are addressed. If preventive measures fail and the patient develops a DVT, several treatment modalities are available. PMID- 8430269 TI - Assessment of disease severity and activity in inflammatory bowel disease. PMID- 8430270 TI - A model of cholestasis in the rat, using a microsurgical technique. AB - An experimental model of extrahepatic cholestasis in the rat, using a microsurgical technique, is described. Sixteen days postoperatively all of the animals (n = 10) were alive and had hepatomegaly, splenomegaly, jaundice, and hyperbilirubinemia. The use of this technique prevents the development of hepatic cysts and other complications inherent in the surgical techniques of cholestasis, such as hepatopneumonic abscesses. PMID- 8430271 TI - Characteristics of teen-age patients with juvenile duodenal ulcer. Relation between inherited hyperpepsinogenemia I and duodenal ulcer. AB - We analyzed environmental factors, family history of peptic ulcer, gastric acid secretion, and serum levels of pepsinogen I (PG I) and gastrin in 56 juvenile patients with duodenal ulcer and 39 normal teenage subjects. Basal acid output and maximal acid output were significantly higher in our duodenal ulcer patients than in controls without ulcer (both, p < 0.01), and patients with duodenal ulcer showed significantly higher serum levels of PG I and gastrin than the controls (both, p < 0.001). There were no significant differences in any environmental factor between the patients and controls. Fifteen of the 17 patients who had one or both parents with hyperpepsinogenemia I had high serum PG I levels. Over half of the duodenal ulcer patients had high serum gastrin levels, irrespective of family history of hypergastrinemia. Our findings suggest that hyperpepsinogenemia I and hypergastrinemia are important characteristics and that genetic background, particularly the inheritance of a gastric mucosal trait expressed as hyperpepsinogenemia I, is frequently involved in the pathogenesis of juvenile duodenal ulcer. PMID- 8430272 TI - The influence of surgically induced acute liver failure on the intestine in the rat. AB - The influence of acute liver failure induced by 90% hepatectomy on the intestine was evaluated in the rat. Small-intestinal mucosal mass decreased 2 h after hepatectomy. Microvillous height decreased significantly from 1 h and on, and villous height and area in the distal small intestine from 2 h after operation. Ninety per cent hepatectomy resulted in a decrease in systemic arterial blood pressure and an increase in portal venous pressure. Subserosal microcirculation and small-arterial circulation in the proximal and distal small intestine and colon decreased significantly after 90% hepatectomy. Overgrowth and colonization of Escherichia coli occurred in the distal small intestine from 1 h and on after hepatectomy. Protein content in enterocytes and bile secretion from the liver remnant were markedly reduced in hepatectomized rats. Thus, the present study shows evidence of alterations in intestinal morphology and function that can contribute to explain the enteric bacterial translocation after surgically induced acute liver failure. PMID- 8430273 TI - Modification of reticuloendothelial function by muramyl dipeptide-encapsulated liposomes in jaundiced rats treated with biliary decompression. AB - Rats with 2 weeks of biliary obstruction, with and without 1 week of concomitant biliary decompression relieving the jaundice, were treated with physiologic saline, free muramyl dipeptide (MDP), placebo liposomes, or liposome-encapsulated MDP. Reticuloendothelial system (RES) function was evaluated by blood clearance of intravenously injected 125I-labelled Escherichia coli. The corrected phagocytic index (alpha) after 1 week of biliary decompression returned to normal levels in animals treated with MDP liposomes, whereas RES function was impaired (P < 0.05) in all other jaundiced and biliary-decompressed groups. In the biliary decompressed, MDP-liposome-treated group, hepatic uptake of radiolabelled bacteria was significantly higher (P < 0.05) and renal entrapment of bacteria was significantly lower (P < 0.05) than in all other jaundiced and biliary decompressed groups. We conclude that treatment with MDP liposomes improves the otherwise impaired RES function in rats with biliary obstruction and biliary decompression. PMID- 8430274 TI - H1 receptor mediates inositol phosphates response to histamine in gastric smooth muscle of guinea pigs. AB - The effect of histamine on [3H]-inositol phosphates (IPs) formation was investigated with [3H]-inositol-labeled gastric smooth-muscle cells in guinea pigs. Histamine (10(-5) M) increased the formation of [3H]-IPs in the muscle cells. The increase was significantly inhibited by pyrilamine (10(-5) M) but not by cimetidine (10(-5) M). The contractile response to histamine was also completely inhibited by pyrilamine but not by cimetidine. Phorbol ester 12 myristate 13-acetate (10 microM) significantly inhibited histamine-stimulated [3H]-IPs formation by 56%, whereas forskolin (10 microM) decreased it by 18%. This study demonstrates that the response of [3H]-IPs formation and contraction to histamine is mediated through H1 receptor, and the formation of [3H]-IPs is negatively regulated by protein kinase C in gastric smooth muscle of guinea pigs. PMID- 8430275 TI - Enterochromaffin-like cells in rat stomach respond to short-term infusion of high doses of cholecystokinin but not to long-term, sustained, moderate hyperCCKemia caused by continuous cholecystokinin infusion or pancreaticobiliary diversion. AB - The histamine-producing enterochromaffin-like (ECL) cells in the oxyntic mucosa are controlled by gastrin. An acute gastrin challenge induces release and accelerated resynthesis of ECL cell histamine. Long-term stimulation with gastrin causes ECL cell hyperplasia. We set out to study whether the ECL cells respond not only to gastrin but also to cholecystokinin (CCK). A wide dose range of gastrin-14 sulfated and -17 non-sulfated and CCK-8 sulfated (CCK-8s) and non sulfated (CCK-8) was infused intravenously to rats for 3 h. The activity of the histamine-forming enzyme was measured at termination of infusion. Gastrins and CCK-8s were equally effective in activating the enzyme, whereas sulfated CCK-8 was notably less potent than the other three peptides. Clearly, the receptor responsible for activation of the ECL cells distinguishes poorly between gastrin 17 and CCK-8s, which is in line with the characteristics of the CCK-B receptor. Moreover, neither the response to gastrin-17 nor that to CCK-8s was affected by concomitant infusion of devazepide (200 micrograms/kg/h), a selective CCK-A receptor antagonist. One group of rats received CCK-8s continuously via a minipump. Another group of rats was subjected to pancreaticobiliary diversion (PBD), which increases the plasma CCK concentration 10- to 20-fold. The rats were killed 7 or 10 weeks later, respectively, and the stomachs were analyzed with regard to mucosal growth and ECL cell hyperplasia. HyperCCKemic rats had increased pancreatic weights but showed no signs of growth stimulation in the stomach and no ECL cell hyperplasia.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430276 TI - Pancreaticobiliary juice releases motilin during phase I of the migrating motor complex in man. AB - The effect of duodenal perfusion with pancreaticobiliary juice on plasma motilin was investigated in eight fasting subjects. The study was undertaken with recording of gastrointestinal motility, to predict spontaneous fluctuations in plasma motilin levels. Duodenal perfusion with pancreaticobiliary juice or saline at a rate of 5 ml/min for 10 min was performed in random order after the second or third activity front. The 30-min integrated plasma motilin response was significantly greater after perfusion with pancreaticobiliary juice: 1.5 (0.7 2.3) nmol x min/l (mean and 95% confidence interval), compared with 1.0 (0.5-1.5) nmol x min/l after saline (p < 0.05). Although perfusion with pancreaticobiliary juice was followed by endogenous plasma motilin peaks after about 20 min in six of eight subjects, the mean interval to the next activity front was not significantly different. In conclusion, duodenal perfusion with pancreaticobiliary juice in phase I of the migrating motor complex releases plasma motilin without affecting the fasting motility pattern. PMID- 8430277 TI - Clinical evaluation of a low junction of the cystic duct. AB - The point of the junction of the cystic duct with the common hepatic duct was studied by means of various preoperative and intraoperative cholangiographic procedures and by gross intraoperative examinations in 468 surgical patients with biliary diseases. The cystic duct entered the hepatic duct at a very low position and was consequently long in 39 patients. The clinical significance of this abnormally low junction of the cystic duct was studied in comparison with 358 patients with gallstones with a normal cystic duct-hepatic duct junction. In the low-junction group with a short common bile duct several complications, including gallstone pancreatitis (7 patients), the Mirizzi syndrome (7), confluence stones (2), gallbladder cancer (3), and congenital dilation of the cystic duct (1), were demonstrated preoperatively. The anomalous junction of the cystic duct with the common bile duct may cause stagnation of bile and/or reflux of pancreatic juice into the bile duct, producing a choledochopancreatic ductal junction and posing difficulties at surgery. PMID- 8430278 TI - Gastroprotective capability of exogenous phosphatidylcholine in experimentally induced chronic gastric ulcers in rats. AB - Phosphatidylcholine (PC) is a main component of the hydrophobic gastric mucosal barrier. Exogenously administered, it prevents acute lesions. We evaluated the gastroprotective capacity of exogenous PC in both acute (ethanol- and indomethacin-induced) and chronic (indomethacin-induced) lesions in rats. Polyunsaturated (PPC) or hydrogenated PC in different concentrations were given intragastrically, before or after the injury factor, in single or repeated doses. Mucosal lesions were significantly reduced by a single dose of PPC, given before or after the injury factor, in both acute models. In the chronic model a single dose of PPC or hydrogenated PC significantly reduced lesions evaluated 6 h after ulcer induction, whereas after 72 h no protective effect was noticed. Repeated doses of PC were ineffective. In conclusion, in acute models exogenous PC reduces lesions in a dose-dependent manner and contributes to the mucosal defense. In chronic models an incomplete and temporary protection might be due to complex pathogenesis that requires activation of all levels in the mucosal defense. Strengthening of only one level was insufficient to restrict injury. PMID- 8430279 TI - Seasonal variation in exacerbations of ulcerative colitis. AB - The monthly distribution of relapses in ulcerative colitis was analysed retrospectively to assess seasonal variation. In 338 patients who were regularly followed up at the colitis clinic, 1013 relapses occurred during the period 1977 to 1988. A peak of relapses occurred in the spring and autumn, with a decrease in the summer; the ratio of the observed to expected number of relapses was 201:255.3 for the summer, 289:252.6 for the autumn, 243:249.8 for the winter, and 280:255.3 for the spring. In contrast, month of diagnosis was evenly distributed without seasonality. The data support the concept that exacerbations in ulcerative colitis are influenced by seasonal factors. PMID- 8430280 TI - [The Neuchatel Society of Dental Medicine. Interview by Kurt Venner]. PMID- 8430281 TI - [The mobile tooth--the theoretical bases and practical implications of a frequent clinical problem]. AB - In the past, especially in the 1970s, tooth mobility has been largely studied by various research groups. Since then, scientific interest in this domain has progressively decreased. However, from a clinical point of view tooth mobility has remained an important concern. This paper aims to review and update basic theoretical notions of tooth mobility after evaluation of recent studies. Finally, clinical implications are discussed. PMID- 8430282 TI - [Overfilling into the mandibular canal as an endodontic complication. A review]. AB - As a consequence of traumatic and toxic injury of the inferior alveolar and the mental nerves, disturbances of sensibility may develop during and after root canal treatment. Several reasons are possible. In most cases, however, such disturbances are the result of root canal filling materials that have been pressed into the mandibular canal. Based on all cases reported in the literature, this review summarizes and discusses symptoms, therapy, prognosis and prophylaxis of such sensibility changes following endodontic overfilling into the mandibular canal. PMID- 8430283 TI - [Marginal defects in amalgam fillings after the use of 2 intermediate substances]. AB - Thirty-six Class V amalgam restorations were placed in 29 patients. Prior to insertion of the amalgam in 12 cavities the walls were treated with Amalgam Liner and in another 12 cavities with Copalite. The remaining 12 cavities were not treated before amalgam insertion. The margin quality was determined quantitatively with the SEM using replicas obtained immediately after the polishing of the restorations and 6 months later. At baseline the cavities treated with Copalite showed a margin quality identical to the untreated cavities. Cavities treated with Amalgam Liner showed an inferior margin quality. After 6 months the margin quality of the three groups showed no statistical differences. With neither of the liners tested was it possible to improve the margin quality of amalgam restorations. PMID- 8430284 TI - [Neural lesions due to endodontics. The indications and therapy]. PMID- 8430285 TI - [Periodontology SGP 1991 (IV)]. PMID- 8430286 TI - [Periodontology SGP 1991 (V)]. PMID- 8430287 TI - [Endodontics: a still important and current topic]. PMID- 8430288 TI - [Plastics as "biological" amalgam alternatives?]. PMID- 8430289 TI - [Max Flesch (1852-1943). A short chapter in the history of the faculty in Bern]. PMID- 8430290 TI - [Hans Rudolf Zangger (1826-1882). Director of the veterinary school in Zurich]. PMID- 8430291 TI - [What is your diagnosis? Decompensating dilated cardiomyopathy]. PMID- 8430292 TI - [Granulated carrots (Daucus carota) in dog nutrition]. AB - In feeding and balance trials with dogs (4) dried, ground carrots were tested in addition (30%) to a basic dry food (rice, corn gluten, fish meal and supplements). The palatability of both mixtures was good: fecal water losses were increased after feeding the carrots. Apparent digestibilities of organic and inorganic matter, especially sodium and potassium (not phosphorus), decreased. Digestibility of carrots was calculated to reach 70% with 1.2 MJ of digestible energy per 100 g dry matter. Reduced fecal pH (6.9-->6.1), higher lactate concentrations and increasing excretion of volatile fatty acids were seen as results of the stimulated bacterial metabolism in the colon. Diurnal pattern of breath hydrogen exhalation was similar in both periods: renal excretions of phenol, indican, and nitrogen were significantly reduced after supplementation with carrots as compared to control period. PMID- 8430293 TI - [A hitherto unknown manuscript of the Mulomedicina Chironis in the Basel University library]. PMID- 8430294 TI - [The very secret, most recently found recipes. The drug book of Js. Jacob Seiler, the veterinarian of Bankon in 1815]. PMID- 8430295 TI - Genes and crime. A US plan to reduce violence rekindles an old controversy. PMID- 8430296 TI - Zinc fingers. PMID- 8430297 TI - Breaching the blood-brain barrier. PMID- 8430298 TI - Basic research (II): Organization. PMID- 8430299 TI - Pitohui: how toxic and to whom? PMID- 8430300 TI - The definition of misconduct in science: a view from NSF. PMID- 8430301 TI - NIH panics over AIDS 'Manhattan Project'. PMID- 8430302 TI - Healy stays, fetal tissue ban goes. PMID- 8430303 TI - Disease gene search goes big science. PMID- 8430304 TI - MSU officials criticized for mishandling data dispute. PMID- 8430305 TI - New technique offers a window on bacteria's secret weapons. PMID- 8430306 TI - Biotechnology. Is Japan a boon or a burden to U.S. industry's leadership? PMID- 8430307 TI - The politics of breast cancer. PMID- 8430308 TI - Epidemiology. Search for a killer: focus shifts from fat to hormones. PMID- 8430309 TI - Zeroing in on a breast cancer susceptibility gene. PMID- 8430310 TI - Cellular changes on the route to metastasis. PMID- 8430311 TI - Current approaches to breast cancer prevention. PMID- 8430312 TI - The development of biological therapies for breast cancer. PMID- 8430313 TI - Demic expansions and human evolution. AB - Geographic expansions are caused by successful innovations, biological or cultural, that favor local growth and movement. They have had a powerful effect in determining the present patterns of human genetic geography. Modern human populations expanded rapidly across the Earth in the last 100,000 years. At the end of the Paleolithic (10,000 years ago) only a few islands and other areas were unoccupied. The number of inhabitants was then about one thousand times smaller than it is now. Population densities were low throughout the Paleolithic, and random genetic drift was therefore especially effective. Major genetic differences between living human groups must have evolved at that time. Population growths that began afterward, especially with the spread of agriculture, progressively reduced the drift in population and the resulting genetic differentiation. Genetic traces of the expansions that these growths determined are still recognizable. PMID- 8430314 TI - The hydrolytic water molecule in trypsin, revealed by time-resolved Laue crystallography. AB - Crystals of bovine trypsin were acylated at the reactive residue, serine 195, to form the transiently stable p-guanidinobenzoate. Hydrolysis of this species was triggered in the crystals by a jump in pH. The hydrolysis was monitored by three dimensional Laue crystallography, resulting in three x-ray diffraction structures, all from the same crystal and each representing approximately 5 seconds of x-ray exposure. The structures were analyzed at a nominal resolution of 1.8 angstroms and were of sufficient quality to reproduce subtle features in the electron-density maps for each of the structures. Comparison of the structures before and after the pH jump reveals that a water molecule has positioned itself to attack the acyl group in the initial step of the hydrolysis of this transient intermediate. PMID- 8430315 TI - Structure of the regulatory complex of Escherichia coli IIIGlc with glycerol kinase. AB - The phosphocarrier protein IIIGlc is an integral component of the bacterial phosphotransferase (PTS) system. Unphosphorylated IIIGlc inhibits non-PTS carbohydrate transport systems by binding to diverse target proteins. The crystal structure at 2.6 A resolution of one of the targets, glycerol kinase (GK), in complex with unphosphorylated IIIGlc, glycerol, and adenosine diphosphate was determined. GK contains a region that is topologically identical to the adenosine triphosphate binding domains of hexokinase, the 70-kD heat shock cognate, and actin. IIIGlc binds far from the catalytic site of GK, indicating that long-range conformational changes mediate the inhibition of GK by IIIGlc. GK and IIIGlc are bound by hydrophobic and electrostatic interactions, with only one hydrogen bond involving an uncharged group. The phosphorylation site of IIIGlc, His90, is buried in a hydrophobic environment formed by the active site region of IIIGlc and a 3(10) helix of GK, suggesting that phosphorylation prevents IIIGlc binding to GK by directly disrupting protein-protein interactions. PMID- 8430316 TI - Patchiness and correlations in DNA sequences. AB - The highly nonrandom character of genomic DNA can confound attempts at modeling DNA sequence variation by standard stochastic processes (including random walk or fractal models). In particular, the mosaic character of DNA consisting of patches of different composition can fully account for apparent long-range correlations in DNA. PMID- 8430317 TI - The skipping of constitutive exons in vivo induced by nonsense mutations. AB - Nonsense mutations create a premature signal for the termination of translation of messenger RNA. Such mutations have been observed to cause a severe reduction in the amount of mutant allele transcript or to generate a peptide truncated at the carboxyl end. Analysis of fibrillin transcript from a patient with Marfan syndrome revealed the skipping of a constitutive exon containing a nonsense mutation. Similar results were observed for two nonsense mutations in the gene encoding ornithine delta-aminotransferase from patients with gyrate atrophy. All genomic DNA sequences flanking these exons that are known to influence RNA splicing were unaltered, which suggests that nonsense mutations can alter splice site selection in vivo. PMID- 8430318 TI - The effect of posttranslational modifications on the interaction of Ras2 with adenylyl cyclase. AB - Ras proteins undergo a series of posttranslational modifications that are critical for their cellular function. These modifications are necessary to anchor Ras proteins to the membrane. Yeast Ras2 proteins were purified with various degrees of modification and examined for their ability to activate their effector, adenylyl cyclase. The farnesylated intermediate form of Ras2 had more than 100 times higher affinity for adenylyl cyclase than for the unprocessed form. The subsequent palmitoylation reaction had little effect. In contrast, palmitoylation was required for efficient membrane localization of the Ras2 protein. These results indicate the importance of farnesylation in the interaction of Ras2 with its effector. PMID- 8430319 TI - Selection of bacterial virulence genes that are specifically induced in host tissues. AB - A genetic system was devised that positively selects for bacterial genes that are specifically induced when bacteria infect their host. With the pathogen Salmonella typhimurium, the genes identified by this selection show a marked induction in bacteria recovered from mouse spleen. Mutations in all ivi (in vivo induced) genes that were tested conferred a defect in virulence. This genetic system was designed to be of general use in a wide variety of bacterial-host systems and has several applications in both vaccine and antimicrobial drug development. PMID- 8430320 TI - CNTF protection of oligodendrocytes against natural and tumor necrosis factor induced death. AB - A proportion of developing oligodendrocytes undergo natural cell death by apoptosis, and mature oligodendrocytes die, either by apoptosis or necrosis, in response to injurious signals such as cytotoxic cytokines and complement. Ciliary neurotrophic factor (CNTF), a trophic factor found in astrocytes in the central nervous system (CNS), promoted the survival and maturation of cultured oligodendrocytes. This trophic factor also protected oligodendrocytes from death induced by tumor necrosis factors (apoptosis) but not against complement (necrosis). These results suggest that CNTF functions in the survival of oligodendrocytes during development and may lead to therapeutic approaches for degenerative diseases of the CNS that involve oligodendrocyte destruction. PMID- 8430321 TI - Inhibition of neural crest cell attachment by integrin antisense oligonucleotides. AB - Neural crest cell interactions with extracellular matrix molecules were analyzed with the use of antisense oligonucleotides to block synthesis of integrin subunits. When added to the culture medium of quail neural crest cells, selected antisense phosphorothiol oligonucleotides reduced the amounts of cell surface alpha 1 or beta 1 integrin subunits by up to 95 percent and inhibited neural crest cell attachment to laminin or fibronectin substrata. Differential effects on specific alpha integrins were noted after treatment with alpha-specific oligonucleotides. Cells recovered the ability to bind to substrata 8 to 16 hours after treatment with inhibitory oligonucleotides. The operation of at least three distinct alpha integrin subunits is indicated by substratum-selective inhibition of cell attachment. PMID- 8430322 TI - A truncated erythropoietin receptor and cell death. PMID- 8430323 TI - Statistical evaluation of DNA fingerprinting: a critique of the NRC's report. PMID- 8430324 TI - Agencies spar over vaccine trial. PMID- 8430325 TI - AIDS research. Reorganization plan draws fire at NIH. PMID- 8430326 TI - Geneticists attack NRC report as scientifically flawed. PMID- 8430327 TI - Cell death studies yield cancer clues. PMID- 8430328 TI - Death gives birth to the nervous system. But how? PMID- 8430329 TI - Genetic models for studying cancer susceptibility. PMID- 8430330 TI - Synaptic vesicle phosphoproteins and regulation of synaptic function. AB - Complex brain functions, such as learning and memory, are believed to involve changes in the efficiency of communication between nerve cells. Therefore, the elucidation of the molecular mechanisms that regulate synaptic transmission, the process of intercellular communication, is an essential step toward understanding nervous system function. Several proteins associated with synaptic vesicles, the organelles that store neurotransmitters, are targets for protein phosphorylation and dephosphorylation. One of these phosphoproteins, synapsin I, by means of changes in its state of phosphorylation, appears to control the fraction of synaptic vesicles available for release and thereby to regulate the efficiency of neurotransmitter release. This article describes current understanding of the mechanism by which synapsin I modulates communication between nerve cells and reviews the properties and putative functions of other phosphoproteins associated with synaptic vesicles. PMID- 8430331 TI - A high-temperature superconducting receiver for nuclear magnetic resonance microscopy. AB - A high-temperature superconducting-receiver system for use in nuclear magnetic resonance (NMR) microscopy is described. The scaling behavior of sources of sample and receiver-coil noise is analyzed, and it is demonstrated that Johnson, or thermal, noise in the receiver coil is the factor that limits resolution. The behavior of superconductors in the environment of an NMR experiment is examined, and a prototypical system for imaging biological specimens is discussed. Preliminary spin-echo images are shown, and the ultimate limits of the signal-to noise ratio of the probe are investigated. PMID- 8430332 TI - Formation of an Fe(III)-tyrosinate complex during biomineralization of H-subunit ferritin. AB - An iron(III)-tyrosinate complex was identified in ferritin by ultraviolet-visible and resonance Raman spectroscopies. Previously, a specific amino acid side chain coordinated to iron in ferritin was not known. Ferritin protein was overexpressed in Escherichia coli from complementary DNA sequences of bullfrog red cell ferritin. The purple iron(III)-tyrosinate intermediate that formed during the first stages of iron uptake was replaced by the amber multinuclear iron(III)-oxo complexes of fully mineralized ferritin. Only the H subunit formed detectable amounts of the iron(III)-tyrosinate complex, which may explain the faster rates of iron biomineralization in H- compared to L-type ferritin. PMID- 8430333 TI - Probing the structure and mechanism of Ras protein with an expanded genetic code. AB - Mutations in Ras protein at positions Gly12 and Gly13 (phosphate-binding loop L1) and at positions Ala59, Gly60, and Gln61 (loop L4) are commonly associated with oncogenic activation. The structural and catalytic roles of these residues were probed with a series of unnatural amino acids that have unusual main chain conformations, hydrogen bonding abilities, and steric features. The properties of wild-type and transforming Ras proteins previously thought to be uniquely associated with the structure of a single amino acid at these positions were retained by mutants that contained a variety of unnatural amino acids. This expanded set of functional mutants provides new insight into the role of loop L4 residues in switch function and suggests that loop L1 may participate in the activation of Ras protein by effector molecules. PMID- 8430334 TI - Editing of transfer RNAs in Acanthamoeba castellanii mitochondria. AB - With the discovery of RNA editing, a process whereby the primary sequence of RNA is altered after transcription, traditional concepts of genetic information transfer had to be revised. The known RNA editing systems act mainly on messenger RNAs, introducing sequence changes that alter their coding properties. An editing system that acts on transfer RNAs is described here. In the mitochondria of Acanthamoeba castellanii, an amoeboid protozoan, certain transfer RNAs differ in sequence from the genes that encode them. The changes consist of single nucleotide conversions (U to A, U to G, and A to G) that appear to arise posttranscriptionally, are localized in the acceptor stem, and have the effect of correcting mismatched base pairs. Editing thus restores the base pairing expected of a normal transfer RNA in this region. PMID- 8430335 TI - Tumor suppression in Xiphophorus by an accidentally acquired promoter. AB - Melanoma formation in the teleost Xiphophorus is caused by a dominant genetic locus, Tu. This locus includes the Xmrk oncogene, which encodes a receptor tyrosine kinase. Tumor induction is suppressed in wild-type fish by a tumor suppressor locus, R. Molecular genetic analyses revealed that the Tu locus emerged by nonhomologous recombination of the Xmrk proto-oncogene with a previously uncharacterized sequence, D. This event generated an additional copy of Xmrk with a new promoter. Suppression of the new Xmrk promoter by R in parental fish and its deregulation in hybrids explain the genetics of melanoma formation in Xiphophorus. PMID- 8430336 TI - Restoration of T cell development in RAG-2-deficient mice by functional TCR transgenes. AB - Introduction of TCR alpha transgene, TCR beta transgene, or both into RAG-2-/ mice differentially rescues T cell development. RAG-2-/- mice have small numbers of TCR-CD4-CD8-(double negative, DN) thymocytes that express CD3 gamma delta epsilon and zeta proteins intracellularly. Introduction of a TCR beta transgene, but not a TCR alpha transgene, into the RAG-2-/- background restored normal numbers of thymocytes. These cells were CD4+CD8+ (double positive, DP) and expressed small amounts of surface TCR beta chain dimers in association with CD3 gamma delta epsilon but not zeta. RAG-2-/- mice that expressed alpha and beta TCR transgenes developed both DP and single positive thymocytes. Thus, the TCR beta subunit, possibly in association with a novel CD3 complex, participates in the DN to the DP transition. PMID- 8430337 TI - Role of the acylated amino terminus of recoverin in Ca(2+)-dependent membrane interaction. AB - Recoverin, a calcium ion (Ca2+)-binding protein of vertebrate photoreceptors, binds to photoreceptor membranes when the Ca2+ concentration is greater than 1 micromolar. This interaction requires a fatty acyl residue covalently linked to the recoverin amino (NH2)-terminus. Removal of the acyl residue, either by proteolytic cleavage of the NH2-terminus or by production of nonacylated recoverin, prevented recoverin from binding to membranes. The acylated recoverin NH2-terminus could be cleaved by trypsin only when Ca2+ was bound to recoverin. These results suggest that the hydrophobic NH2-terminus is constrained in Ca(2+) free recoverin and liberated by Ca2+ binding. The hydrophobic acyl moiety of recoverin may interact with the membrane only when recoverin binds Ca2+. PMID- 8430338 TI - Laser-enhanced NMR spectroscopy: theoretical considerations. PMID- 8430339 TI - Calcific tendinitis: a review of the usual and unusual. PMID- 8430340 TI - Skeletal manifestations of lysinuric protein intolerance. A follow-up study of 29 patients. AB - Lysinuric protein intolerance (LPI) is an autosomal recessive disease caused by defective transport of the cationic amino acids lysine, arginine, and ornithine at the cell membrane. About 80 patients with LPI have been described worldwide, almost half of them in Finland. The symptoms appear in early childhood as a failure to thrive, growth retardation, muscular hypotonia, and episodes of stupor after protein-rich meals. Twenty-nine Finnish patients (current median age 24.8 years, range 3.7-47.9 years) over a mean follow-up time of 18.1 years (range 1.2 27.2 years) had 57 fractures after minor trauma, mostly in childhood. Their 440 skeletal radiographs showed severe osteoporosis (13/29), controversially abnormal thickening of cortex of the metacarpals (7/29), or thin cortices of the long bones (5/29), endplate impression of vertebrae (8/29), rickets-like metaphyses (2/29), or early destruction of cartilage (3/29). Skeletal maturation was delayed by 1-5 years in 23 of 24 patients. There was no correlation between fracture incidence, radiological bone structure, and delayed skeletal maturation. PMID- 8430342 TI - Osteoarthritis of the metacarpophalangeal joints: the relation between ray prevalence, trauma, and utilisation. AB - The gradient in prevalence of idiopathic osteoarthritis of the metacarpophalangeal (MCP) joints from the index to the little finger was used to test three theories. Trauma was correlated by analysis of fracture incidence in the four rays in 142 patients. Correlation with utilisation of arc was assessed by goniometric measurement of mean MCP flexion in five hand positions in a sample of 200 activities, and recording utilisation of these positions by behavioural observation techniques in the course of 2831 actions, 2746 domestic and 85 nondomestic. Both correlations were negative. The hypotheses of excessive loading and incomplete use were compared and tested against external evidence. It is suggested that the gradient in MCP prevalence of osteoarthritis is consistent with a bifactorial aetiology, with site and incidence determined by incomplete use, and with the rate of progress by cartilage loading. PMID- 8430341 TI - CT evaluation of primary epiphyseal bone abscesses. AB - We reviewed the clinical, radiographic, and computed tomographic (CT) findings in eight children with a histologically proven diagnosis of epiphyseal or apophyseal osteomyelitis. In all cases the femur was involved: in five the osteomyelitis was localized in the femoral condyle, in two it was in the greater trochanter, and in one it was in the femoral head epiphysis. In four of the six cases of epiphyseal involvement there was associated joint effusion or septic arthritis. CT examination may demonstrate a serpentine tract, a sequestrum, cortical destruction or adjacent soft tissue swelling and can differentiate osteomyelitis from other epiphyseal lucent lesions, particularly chondroblastoma and osteoid osteoma. CT yielded important new diagnostic information in seven of the eight patients, failing to do so in only one. In one case, CT showed a wooden splinter in an abscess cavity, which had been mistaken for a sequestrum. When combined with accurate clinical and laboratory information and good quality plain radiographs, CT can lead to an early diagnosis of epiphyseal infection. Early diagnosis helps avoid delays in initiating antibiotic or surgical treatment caused by the unusual (epiphyseal or apophyseal) location of the bone abscess. PMID- 8430343 TI - Ultrasound examination for measurement of femoral anteversion in children. AB - Femoral anteversion (AV) angles were determined by ultrasound and biplanar radiography in 63 children aged 3-11 years. With the patient supine, knees flexed 90 degrees, and legs kept vertical, only one ultrasound scan of the proximal femur was needed. The anterior tangent of the femoral head and the greater trochanter was used as the reference line. The transducer was tilted until this tangent appeared horizontal on the monitor screen, and the angle of tilt, which represents the AV angle, was measured with an attached clinometer. There was a high correlation between ultrasound and radiographic results (r = 0.71). However, the AV angles appeared an average of 5.5 degrees larger by ultrasound. Thus, a correction factor of 5 degrees should be subtracted from the angle measured by ultrasound to obtain the actual AV angle in children. Because ultrasound does not require ionizing radiation, and because reliable results are obtained, our technique is recommended for screening of children with rotational disorders of the femur. PMID- 8430344 TI - A sign for the early detection of medullary sclerosis. AB - The radiographic image of a normal bone has a continuous radiopaque boundary of variable thickness and a relatively radiolucent interior. Normally, the endosteal edge of this boundary line is well defined. However, this edge may be obscured in diseases that induce medullary new bone formation. The use of this sign for the early detection of medullary sclerosis is demonstrated by specific examples. PMID- 8430345 TI - MR appearance of axial melorheostosis. PMID- 8430346 TI - Case report 753. Tumoral amyloidosis at multiple sites. PMID- 8430347 TI - Case report 766. Pseudomalignant osseous tumour of the soft tissue. PMID- 8430348 TI - Case report 767. Osteosarcoma arising in bone infarction. PMID- 8430349 TI - Case report 768. Eosinophilic granuloma of the cervical spine. PMID- 8430350 TI - Case report 769. Fibrous dysplasia. PMID- 8430351 TI - Case report 770. Chronic subacromial bursitis with massive formation of rice bodies. PMID- 8430352 TI - Detection of Chlamydia trachomatis in men. Sensitive tests for sensitive urethras. AB - The value of examining the centrifuged deposit from a first pass urine sample by an amplified enzyme-linked immunosorbent assay (ELISA) to detect Chlamydia trachomatis was assessed. The results were compared with those obtained by examining a urethral smear with a direct fluorescent antibody (DFA) test (MicroTrak; Syva, Palo Alto, CA). When the first pass urine sample yielded positive results for the IDEIA, but results of the DFA test of the corresponding urethral sample were negative, the remaining centrifuged deposit from the urine sample was examined by the DFA test. If results of this DFA test were positive, the IDEIA result was regarded as a true positive. Paired samples were collected by nursing staff from 190 men attending a clinic for sexually transmitted diseases (STDs). The quality of sampling thus reflected routine practice. The sensitivity of the IDEIA using first pass urine samples was 90% (37/41), and the sensitivity of the DFA test of urethral samples was 83% (34/41). The adoption of testing of first pass urine samples for the detection of C. trachomatis in STD clinics will not lead to a decrease in sensitivity for detecting C. trachomatis and will be more acceptable to patients than urethral swabbing. PMID- 8430353 TI - Physical and psychological effects of anogenital warts on female patients. AB - Symptoms and psychological reactions of 82 patients with anogenital warts were recorded in a descriptive study at the beginning and after treatment. A majority of the patients had substantial and longstanding symptoms like local pain, tenderness, and discharge. Anxieties about cancer and cure were common. With a mood adjective checklist, the patients were compared to 58 referents, matched for background variables. There were significant differences in experiences of pleasantness, activation, and social orientation, which suggest a conspicuous emotional effect of the disease. The clinical picture and pathology of anogenital warts are well known, but the profound impact, hitherto unknown, of the disease on women's physical and psychological well-being should also be taken into account when designing health care and prevention. PMID- 8430354 TI - Factors associated with condom use in a high-risk heterosexual population. AB - The use of condoms has been advocated as a means of preventing the transmission of the human immunodeficiency virus and other sexually transmitted agents. To better understand factors that may influence condom use, 300 heterosexuals were enrolled in a cross-sectional study of patients attending San Francisco's only public sexually transmitted disease clinic. Interviewer-administered questionnaires were conducted. Condom use at last sexual intercourse was examined by logistic regression analysis. Men who used drugs or alcohol at last intercourse and whose partners did not want to use condoms were less likely to have used them; women who were black or Hispanic, who reported difficulty getting their partners to use condoms, or who reported that condoms decrease sexual pleasure also were less likely to have used them. Efforts to increase condom use in this population should target minorities, assist women to negotiate their use, emphasize the dangers of using alcohol and other drugs with sex, and address the perception that condoms interfere with sexual pleasure. PMID- 8430355 TI - Decreased prevalence of Chlamydia trachomatis infection associated with a selective screening program in family planning clinics in Wisconsin. AB - The effectiveness of selective screening for control of Chlamydia trachomatis is unknown. In 1986, a statewide screening program began in family planning clinics in Wisconsin after the prevalence of infection among women was found to be 10.7% in four nonurban clinics and 13.7% in an urban Milwaukee clinic. In 1990, endocervical specimens were obtained from 1,757 women attending these same clinics; 5.2% of women in the non-urban clinics and 6.9% in the Milwaukee clinic tested positive for C. trachomatis. Prevalence of infection had decreased similarly (by 53% overall) in both high- and low-risk groups in all five clinics. Although reported condom use increased from 16% to 31%, most other demographic and behavioral risk factors for infection did not significantly change; in contrast, the prevalence of clinical signs of infection decreased. The percentage of infections identified by selective screening criteria decreased from 77% to 55%. Selective screening and attendant activities, as well as an increase in condom use, were associated with a decrease in prevalence of C. trachomatis infection in this population. PMID- 8430356 TI - Virucides in prevention of HIV infection. Research Priorities. World Health Organization Working Group on Virucides. AB - Vaginal spermicides have microbicidal properties that suggest their usefulness in helping to protect against human immunodeficiency virus (HIV) and other sexually transmitted infections. Laboratory, animal, and clinical investigations also support this notion. Because of the importance of identifying additional methods to protect against these infections, however, better information is needed on these female-controlled methods. A consistent set of laboratory standards is needed to evaluate the in vitro activity of existing and future virucides. Further evaluation of such areas as the actions of virucides and vehicles on different anatomical sites and safety studies to better judge determinants of toxicity are needed. Clinical studies might compare efficacy of spermicides and condoms in prevention of HIV and other sexually transmitted diseases as well as investigate psychosocial considerations that determine suitable candidates for vaginal virucides, how they are used, and how their use might be improved. PMID- 8430357 TI - Chlamydial infection and sexual behavior in young pregnant teenagers. AB - Chlamydia trachomatis infection is one of the most common sexually transmitted diseases (STD) among adolescents and is one of the most common causes of perinatal infection. We have screened 267 young nulliparous adolescent girls (12 17 years old) for cervical C. trachomatis infection during their pregnancy using tissue culture methods. The population was a cohort of pregnant teenagers who had limited sexual exposure (mean duration of sexual activity, 14.3 months, mean number of lifetime partners, 2.0 +/- 1.5). The initial screening was done at enrollment to a prenatal care clinic, and the third trimester examination at 32 to 36 weeks of gestation. According to clinical indications, additional interim STD examinations were carried out. The cervical C. trachomatis infection rate was 18.7% (50/267) at intake and 7.9% (14/178) in the third trimester. Three of the 40 interim STD examinations also were positive for cervical C. trachomatis infection. Prenatally, 64 of 267 (24.0%) girls were positive for C. trachomatis at one or more times. The repeat culture positive rate for C. trachomatis was 7.8% (17/218), and 14 of those that were negative on the initial screening were positive on the subsequent screening. One or more types of treatable STDs (chlamydial, gonorrhea, syphilis, or trichomonal) were identified in 38.6% (103/267) of the study population during pregnancy. These findings suggest that high-risk sexual behavior may continue in teen pregnancy, and repeated prenatal chlamydial and other STD screening and counseling are indicated in this population. PMID- 8430358 TI - Transport media for Haemophilus ducreyi. AB - Stuart's, Amies' and four thioglycollate/hemin-based media containing, respectively, selenium dioxide and albumin (SA); selenium dioxide, L-glutamine and albumin (SGA); selenium dioxide and L-glutamine (SG); and L-glutamine and albumin (GA); were evaluated as transport media for Haemophilus ducreyi in a simulated, laboratory-based study. No transport medium was able to maintain the viability of H. ducreyi for more than 24 hours at room temperature. Amies' transport medium, however, maintained viability of H. ducreyi for up to 3 days and all the newly formulated transport media maintained viability of H. ducreyi for more than 4 days at 4 degrees C. Subsequently, swabs were taken from cases of genital ulcerations with a diagnosis of chancroid, plated directly onto selective media, and held at 4 degrees C in the two most efficient transport media (SGA and GA) for various periods of time. On direct plating, H. ducreyi was recovered from the genital ulcerations of 110 of 156 (71%) patients. The rates of recovery of H. ducreyi from transport media stored at 4 degrees C decreased with time. Although recovery was enhanced at day 4 (117/110 [106%]) using both SGA and GA, a recovery rate of 37/41 (90%) was obtained after storage in GA for 1 week. PMID- 8430359 TI - Factors correlated with homosexually acquired human immunodeficiency virus infection in the era of "safer sex". Was the prevention message clear and well understood? Alain Brugeat Physician Group. AB - A cross-sectional survey was conducted between November 1986 and January 1988 among 246 homosexual/bisexual patients by consulting physicians promoting human immunodeficiency virus (HIV) infection prevention, to determine factors correlated with HIV infection a few years after the launch of safer sex recommendations. After adjustment for numbers of sexual partners and frequency of unprotected receptive anal intercourse, seropositive subjects, compared to seronegative ones, had significantly higher frequencies of: (1) receptive anal intercourse using condoms and extraneous lubricants, (2) anorectal douching before or after intercourse, (3) past history of syphilis, and (4) nitrite inhalations. The first three factors remained significant after multivariate analysis. Eighty-three percent of the subjects practicing receptive anal intercourse with condoms plus lubricants used inappropriate lubricants. Some factors identified in our study are well established risk factors for homosexually acquired HIV infection, suggesting that safer sex recommendations still are not followed by all. Our results also elicit additional factors that independently increase the risk. Two of them, extraneous lubrication of the condom for anal receptive intercourse and anorectal douching, may result from a misunderstood notion of "safer sex," or from practices thought by mistake to protect against HIV. PMID- 8430360 TI - Current management of malignant melanoma. PMID- 8430361 TI - Parathyroid autotransplantation. AB - It has been clearly shown clinically that parathyroid tissue can be successfully autotransplanted and even allotransplanted if the host is immunosuppressed. Engraftment is almost always successful; however, if abnormal tissue has been transplanted, its function will continue to be abnormal if the same intrinsic (e.g., primary parathyroid hyperplasia) or extrinsic (renal osteodystrophy) stimulation existing before grafting continues in the postoperative period. In these patients the secretion of parathyroid hormone from the grafted parathyroid tissue can be shown to progressively increase with time. Although parathyroid autotransplantation is most frequently performed clinically for renal osteodystrophy, there is controversy about the operation, and some surgeons prefer the technique of 3 1/2 gland parathyroidectomy. Because of the generally good results with total parathyroidectomy and autotransplantation in patients with primary parathyroid hyperplasia, this procedure seems to be the operation of choice in this clinical setting. The clearest indication for parathyroid autotransplantation is in patients with radical operations on the thyroid gland or other head and neck organs where the parathyroids have been damaged or their viability is questioned. PMID- 8430362 TI - Postoperative radiotherapy for carcinoma of the esophagus: a prospective, randomized controlled study. AB - METHODS: A prospective, randomized controlled study of radiotherapy after resection of esophageal carcinoma was carried out in 130 patients. Patients were stratified according to whether the resection was curative or palliative and were then randomized to receive postoperative radiotherapy or no additional treatment. Sixty patients underwent curative resection; 30 each were randomized into the radiotherapy group (CR + R) and the control group (CR). Seventy patients underwent palliative resection; 35 each were randomized into the radiotherapy group (PR + R) and the control group (PR). RESULTS: No complications occurred while the patients were undergoing radiotherapy treatment. On follow-up, complications in the intrathoracic stomach occurred in 24 patients (37%) who underwent radiotherapy compared with four patients (6%) in the control group (p < 0.0001). Seventeen of these 24 patients in the radiotherapy group had gastric ulceration and there were five deaths as a result of bleeding. Local recurrence developed significantly less frequently in the PR + R group compared with the PR group (seven patients [20%] vs 16 patients [46%]; p = 0.04); no difference was observed between CR + R and CR groups (10% and 13%, respectively). Intrathoracic recurrence occurred in fewer patients in the radiotherapy groups (CR + R and PR + R) compared with the control groups (CR and PR) (four patients vs 15 patients; p = 0.01). In patients with residual tumor in the mediastinum after resection, two (7%) of 29 patients who underwent radiotherapy died of tracheobronchial obstruction, compared with nine (33%) of 27 patients in the control groups (p = 0.03). No difference in local recurrence was observed for extrathoracic or anastomotic recurrence. Distant metastasis developed in 12 patients (40%) in the CR + R group, nine patients (30%) in the CR group (p = 0.59), 24 patients (69%) in the PR + R group, and 18 patients (51%) in the PR group (p = 0.22). The time of onset of metastasis was 5.1 months for the PR + R group, which was shorter than the 8.5 months for the PR group (p = 0.05). The time of onset of metastasis was similar for the CR + R and CR groups (9.9 months and 11.0 months, respectively; p = 0.76). The overall median survival of patients after postoperative radiotherapy (CR + R and PR + R) was 8.7 months, which was shorter than the 15.2 months for the control groups (CR and PR) (p = 0.02). CONCLUSIONS: The shorter survival of patients who underwent postoperative radiotherapy was the result of irradiation-related death and the early appearance of metastatic diseases. The role of postoperative radiotherapy is therefore limited to a specific group of patients with residual tumor in the mediastinum after operation, for whom radiotherapy can significantly reduce the incidence of local recurrence obstructing the tracheobronchial tree. PMID- 8430363 TI - Education about death and dying during the clinical years of medical school. AB - BACKGROUND: Although there has been a dramatic increase in education about death and dying in medical school curricula, the physician's interaction with terminally ill patients and their families still causes concern. The purpose of our study was to determine the impact of the third-year clerkship on education of medical students about death and dying. METHODS: From August 1, 1988, to August 1, 1990, a questionnaire concerning the care of terminally ill patients was distributed to all students completing the third-year clinical clerkship at our medical school. RESULTS: One hundred and eighty questionnaires were distributed, of which 106 were returned, yielding a response rate of 59%. All students had cared for a terminally ill patient during their third year. Forty-four (41%) students responding had never been present when an attending physician talked with a dying patient, and 37 (35%) had never discussed with an attending physician how to deal with a terminally ill patient. During the surgical clerkship 77 (73%) students had never been present when a surgeon had to tell the family of a patient bad news after surgery, and 90 (85%) had never been present when an attending surgeon had informed a family that their relative had died. Despite the fact that the curriculum addresses the stages of death and dying, almost half of the students could not remember these. When they were discharging a terminally ill patient home, one third of students could not identify problems that would be encountered by the family in caring for the patient. CONCLUSIONS: Fifty-seven (54%) felt that they were poorly equipped to deal with terminally ill patients on graduation from medical school, and 91% welcomed the opportunity to be educated in this area during the clinical years. PMID- 8430364 TI - Thyroid carcinoma with tracheal or esophageal involvement: limited or maximal surgery? AB - This retrospective study was undertaken to evaluate the prognosis of and to determine optimal therapy for thyroid carcinoma adhering to or invading the trachea or esophagus. In our series of 412 operated thyroid cancers, there were 45 patients including 20 with adherences to the trachea or esophagus that were dissected free by sharp dissection (group 1), six patients with invasion of the trachea or esophagus who underwent total resection followed by radioiodine and external-beam irradiation (group 2), and 19 patients with invasion of the trachea or esophagus that had been incompletely resected (group 3). There were no major complications. Survival or disease-free unrelated deaths were recorded in 80%, 100%, and 16% of patients in groups 1, 2, and 3, respectively. The three patients with anaplastic carcinoma in group 1 are free of disease 3, 6, and 7 years after operation, respectively. Combined with those in the literature, our data indicate that (1) adherences to the trachea or esophagus are not necessarily associated with poor prognosis and should be treated with aggressive surgery even in anaplastic carcinoma, (2) invasion of the trachea or esophagus must be treated whenever possible by total resection followed by radioiodine and external-beam irradiation, (3) a two-stage operation should be considered when optimal conditions are not available initially, and (4) cure may be obtained with complementary radioiodine and external-beam irradiation after incomplete resection of papillary carcinoma. PMID- 8430365 TI - Tumor necrosis factor-alpha inhibits in vivo collagen synthesis. AB - In this study we sought to determine the in vivo role of tumor necrosis factor alpha (TNF-alpha) at the wound-healing site. In vivo abrogation of endogenous TNF alpha activity in experimental wounds by administration of anti-murine TNF-alpha rabbit serum resulted in a significant 77.5% increase in wound collagen deposition, as assessed by wound sponge granuloma hydroxyproline content. Administration of pharmacologic doses of recombinant murine TNF-alpha into subcutaneously inserted polyvinyl alcohol sponges resulted in an increase in collagen deposition (1594 +/- 117 vs 1014 +/- 49 and 1588 +/- 135 vs 1014 +/- 49 micrograms/100 mg sponge, for TNF-alpha in situ administration at a dose of 0.05 and 0.5 micrograms, respectively). This effect could be abolished by the simultaneous systemic treatment of the animals with the antiinflammatory drug indomethacin. The data suggest that the enhanced collagen deposition after TNF alpha administration is a consequence of a nonspecific inflammatory activity that indirectly promotes collagen synthesis. The data also support the hypothesis that endogenous wound TNF-alpha down-regulates collagen synthesis during normal wound healing. PMID- 8430366 TI - Management of sudden profuse bleeding from varicose veins. AB - Varicose vein bleeding of the lower extremities is an unusual but pressing indication for treatment and can be lethal. This series reviews operative and injection treatment of such veins in patients with bleeding. During a 49-month period 14 patients (eight men and six women) with a mean age of 62.1 years (range, 23 to 93 years) were seen after venous bleeding related to varicosities of the lower extremity. They described between one and five episodes of bleeding (mean, 2.4), but only one patient required transfusion. The site of bleeding was the lower calf or foot in 11 and affected the thigh in three patients. One was in the third trimester of pregnancy. Nine patients had lesions involving clustered small 1 mm or less diameter varicose veins, whereas five had large diffuse varicose vein formation. None had evidence of coagulopathy, and the inciting episode was either unknown or related to minor trauma. Treatment of eight patients with small-diameter veins was instituted with 0.2% sodium tetradecyl injection with a 30-gauge needle, thrombosing veins within 5 cm of the bleeding focus. A total of 13 ml solution (1.0 to 27 ml) per patient was used during two or three treatment periods (mean, 2.5 treatment periods) spaced 2 weeks apart. Six patients were treated by means of standard vein-stripping techniques or local branch removal at the bleeding site. Effective thrombosis was achieved in all eight patients with small (less than 1 mm) varicose veins treated with sclerosis. In five patients who underwent surgery there was effective resolution of the hemorrhage. One patient with small-diameter varicose veins clustered about the ankle underwent operative treatment and had venous stasis ulceration requiring compression dressings for healing. In all 14 cases control of bleeding was obtained, with follow-up to 49 months (mean, 21.3 months); one had rebleeding from a site 32 cm remote from the original injected area 11 months after treatment. Bleeding from small-diameter varicose veins of the lower extremity can be controlled effectively by sclerosing techniques with sodium tetradecyl. Larger veins are managed with operative removal. Combining these techniques provides efficient management of often-elderly patients, many in an outpatient setting. PMID- 8430367 TI - Reperfusion mucosal damage after complete intestinal ischemia in the dog: the effects of antioxidant and phospholipase A2 inhibitor therapy. AB - In a recent study, reperfusion mucosal injury was demonstrated in a rat model of total ischemia if venous congestion was avoided. The aims were to examine the possibility of reperfusion damage in a canine model involving 2 hours of complete segmental ischemia and to investigate the effects of antioxidant therapy or pretreatment with nonspecific phospholipase A2 inhibitors on postocclusive mucosal changes. Tissue samples were evaluated histologically in a blind manner, according to a 0 to V grade scale. The degree of mucosal damage was statistically significantly increased during the 30-minute reperfusion period. Similarly, 2 hours of total ischemia followed by 30 minutes of reperfusion produced significantly more tissue lesions than did 2 1/2 hours of ischemia without reperfusion. Oral allopurinol pretreatment supplemented by an intravenous dose, or oral allopurinol in combination with a superoxide radical scavenger, resulted in a significant amelioration of postischemic histologic changes. Pretreatment with a nonspecific phospholipase A2 inhibitor (methylprednisolone, dexamethasone, or quinacrine) was ineffective in diminishing the reperfusion injury in either case. The results suggest that reperfusion injury may develop even after complete intestinal ischemia, and this damage can be attenuated by inhibiting the capacity of xanthine oxidase to generate reactive oxygen intermediates. PMID- 8430368 TI - Recent advances in the management of Fournier's gangrene: preliminary observations. AB - Twenty consecutive cases of Fournier's gangrene managed conservatively with systemic antibiotics and topical application of unprocessed honey (group A) were compared with 21 similar cases of Fournier's gangrene managed by the orthodox method (group B) during the same period. Group A received oral amoxicillin/clavulanic acid and metronidazole in addition to daily topical application of honey to the gangrenous scrotum, whereas group B underwent wound debridement, wound excision, secondary suturing, and in some cases scrotal plastic reconstruction in addition to receiving a mixture of systemic antibiotics dictated by culture and sensitivity results. The organisms cultured in both groups were similar. Even though the average duration of hospitalization was slightly longer in group A (4.5 weeks) as opposed to group B (4 weeks), topical application of honey showed distinct advantages over the orthodox method. Three deaths occurred in group B, whereas no deaths occurred in group A. The need for anesthesia and expensive surgical operation was obviated. Response to treatment and alleviation of morbidity were faster in group A. Honey may revolutionize the treatment of this dreadful disease by reducing morbidity and mortality. PMID- 8430369 TI - Pretreatment of fetal porcine pancreas in culture with nicotinamide accelerates reversal of diabetes after transplantation to nude mice. AB - Successful transplantation of fetal pancreatic beta-cells to recipients with diabetes requires that differentiation of the immature beta-cells is induced. In this study isletlike cell clusters (ICC) were produced from fetal porcine pancreas in tissue culture, in medium RPMI-1640 supplemented with 10% human serum and in the presence or absence of 10 mmol/L nicotinamide. Light microscopy of immunostained ICC on day 4 of culture showed that nicotinamide caused a more than doubling in the frequency of insulin-positive cells, whereas no stimulatory effect by nicotinamide on DNA replication was found. Transplantation of ICC into nude mice with alloxan diabetes revealed that ICC formed in nicotinamide normalized the hyperglycemia faster than did control ICC (3 to 4 weeks compared with 6 to 8 weeks). The DNA content of the transplanted ICC increased two to three times over an 8-week period, whereas the insulin content increased 100 fold. The total insulin, total DNA content, and the insulin concentration of the grafts were significantly higher in the nicotinamide grafts in comparison with control grafts. In fact, the insulin concentration in the nicotinamide grafts was almost identical to that normally observed in fully differentiated mouse islets. Furthermore, perfusion in vitro of the graft-bearing kidneys 18 weeks after transplantation showed that the total amount of insulin released from the nicotinamide grafts on stimulation with glucose was more than five times higher than the amount from the control grafts. The perfusion of grafts from the nicotinamide group revealed a marked biphasic insulin response to glucose, which was less obvious in the control grafts. We conclude that nicotinamide induces beta-cell differentiation in porcine ICC and that this effect is beneficial when such explants are used for transplantation to recipients with diabetes. PMID- 8430370 TI - Bilateral lobectomy excluding the caudate lobe for giant mesenchymal hamartoma of the liver. AB - A girl underwent partial resection of the liver for mesenchymal hamartoma twice, at 7 months and again at 3 years of age. When the patient was 16 years of age, the mass increased rapidly in size. The patient was hospitalized and studied by ultrasonography, computed tomographic scans, and arterial angiography. The tumor occupied the right and left lobes of the liver, but the hypertrophic caudate lobe in which no tumor was found could provide satisfactory liver function, so the tumor was extirpated by bilateral lobectomy, with preservation of the caudate lobe. Histologic examination of the operative specimen led to the diagnosis of typical mesenchymal hamartoma of the liver. Malignancy was not found. About 30 cases of this disorder have been reported in Japan, and about 100 cases have appeared in the literature in English. To the best of our knowledge, no earlier reports of bilateral lobectomy have been made. PMID- 8430371 TI - Adenocarcinoma arising within a colonic diverticulum: report of two cases and review of the literature. AB - The first reported cases of colonic carcinoma arising within a diverticulum are documented. Although colonic diverticulitis and cancer are common diseases in patients over 60 years of age, cancer arising within a diverticulum is rare. Only close histopathologic scrutiny can differentiate inflammatory changes from neoplasia. Because colonic diverticula are characteristically thin-walled, cancers arising within diverticula may easily penetrate the serosa and first be diagnosed at an advanced stage despite their small size. PMID- 8430372 TI - Bilateral internal carotid artery agenesis: a case study and review of the literature. AB - Bilateral internal carotid artery agenesis is a rare lesion, with only 18 cases previously reported. Blood supply to the anterior cerebral circulation is most commonly through enlarged basilar and posterior communicating arteries. Occasionally collateral flow is through abnormal transsellar anastomoses or anastomoses between the external carotid and intracranial systems. Associated intracranial aneurysms occur in 25% of patients, accounting for a significant incidence of intracranial hemorrhage as the initial symptom. Diagnosis is best made by cerebral arteriography with computed tomography to verify the congenital nature of the abnormality by demonstrating the absence of carotid canals. This is the second case to begin with transient ischemic attacks suggestive of carotid territory ischemia but originating from the vertebral system. Angiographic findings included absent internal carotid arteries, small common carotid arteries, and bilateral high-grade stenoses at the origins of large vertebral arteries. This is the first such case to be treated with reimplantation of the vertebral artery with resolution of symptoms. PMID- 8430373 TI - Curious interaction of bugs and bees. PMID- 8430374 TI - Re: Prognostic factors in medullary carcinoma of the thyroid. PMID- 8430375 TI - Use of polypropylene mesh to repair contaminated abdominal wounds in rats. PMID- 8430376 TI - Knowledge of UR law gives physicians advantage during interviews. PMID- 8430377 TI - Failure to report child, elder abuse is criminal offense in Texas. PMID- 8430378 TI - POEP makes strides in cancer education. PMID- 8430379 TI - Report urges physician, school involvement against substance abuse. PMID- 8430380 TI - Agenda on AIDS. PMID- 8430381 TI - Tobacco, alcohol questions now appear on death certificates. PMID- 8430383 TI - Court decisions demonstrate new fairness. PMID- 8430382 TI - Physicians not alone in battle against incest. PMID- 8430384 TI - Digitized doctoring. A healthy mix of medicine and computers. AB - Computers, these days, are almost smaller than a bread box. They sit unobtrusively atop a desk, silently awaiting the next command. And yet, for such a small package, these microelectronic-based, information-processing devices have profoundly affected mankind in the second half of the 20th century. Computer technology has radically altered how -- and how fast -- the world changes. PMID- 8430385 TI - New Medicaid payment system brings more physicians into program. PMID- 8430386 TI - CLIA inspection of physician office labs to begin this year. PMID- 8430387 TI - Work satisfaction in the lives of physicians. AB - In a climate that many practitioners find increasingly oppressive, data suggesting the most and least satisfying aspects of medical practice are important. In a joint effort, the Dallas County Medical Society, Timberlawn Psychiatric Hospital, and Timberlawn Psychiatric Research Foundation explored levels of work satisfaction for their relationships with marital satisfaction and psychiatric symptoms as reported by physicians and dentists and their spouses. Most physicians and dentists rate various aspects of direct patient care and service to humanity as highly satisfying aspects of professional life. Physicians highly satisfied with their work are more likely to report high satisfaction in their marriages and fewer psychiatric symptoms. Most spouses of physicians report high levels of marital satisfaction, and high agreement exists between the marital satisfaction ratings of physicians and their spouses. Physicians and dentists are quite similar on the wide range of professional and personal variables surveyed. PMID- 8430388 TI - Work stress in the lives of physicians. AB - The work stress of physicians is considered high, and many are contemplating leaving the profession. Documenting the specific aspects of practice found to be most stressful is important, and useful insights may be obtained by contrasting physicians who report high and low levels of work stress. Contrasting the work stress of physicians and dentists can be useful for both professions. Accordingly, we surveyed 3156 members of the Dallas County Medical Society and 1273 members of the Dallas County Dental Society plus a large group of their spouses. Although the level of work satisfaction is the most powerful predictor of work stress, other significant predictors are younger age, long working hours, and solo practice. Physicians who report high levels of work stress also report lower levels of marital satisfaction and a higher prevalence of psychiatric symptoms. Dentists are much like physicians in their reports of overall work stress, and the similarities and differences regarding specific stressors suggest the two professions are more alike than different in reporting the stresses of professional practice. PMID- 8430389 TI - Resuscitation devices should be as available as fire extinguishers. PMID- 8430390 TI - Education is key to firearm safety. PMID- 8430391 TI - [Environmental protection is health protection]. PMID- 8430392 TI - [Do we need a specialty of adolescent medicine in Norway?]. PMID- 8430393 TI - [Transgenic mice, basic research and source of wishes]. PMID- 8430394 TI - [Blood pressure measurements]. PMID- 8430395 TI - [Prevalence of hospital infections in Norwegian somatic hospitals]. AB - A nation-wide survey of the prevalence of hospital-acquired infections was carried out on 25 April 1991. The purpose of the study was to assess the size of the problem, to find out in which medical disciplines they occur and what types of infections dominate, and to motivate hospital personnel for training in infection control. The survey included 77 somatic hospitals with a total of 15,160 patients. 4,418 patients had undergone surgery during their stay in hospital. 1,063 were newborns. On the day of the survey 976 clinically manifest infections were recorded. This gave a total prevalence rate of 6.4%. At county level the prevalence rates varied from 3.8 to 8.9%. The prevalence rate for surgical wound infections was 3.7%. Urinary tract infections accounted for 33.6% of the total infections, followed by lower respiratory tract infections (16.8%), and surgical wound infections (16.6%). The study indicates in which medical disciplines the problems of infection are most serious, and where the control should be strengthened. It also provides a good basis for further detailed studies of hospital-acquired infections. PMID- 8430396 TI - [Course of obstructive lung diseases]. AB - Obstructive lung disease can be divided into bronchial asthma and chronic obstructive lung disease. Forced expiratory volume for one second (FEV1) is the most commonly used variable to follow the course of obstructive lung disease. In healthy who have never smoked, FEV1 reaches a plateau phase at the age of 20-35 years, before starting to decrease with increasing age. In subjects with obstructive lung disease this plateau phase does not exist and the decline in FEV1 is steeper than in healthy subjects. Asthmatics who stop smoking, double the probability of remission of the disease. Remission of occupational asthma is dependent on early diagnosis of the disease. The disease seems to follow a more benign course in asthmatics with an allergic disposition than asthmatics without. The effect of a bronchodilator and antiinflammatory treatment on the course of obstructive lung disease is not known. Long-term oxygen therapy is the only treatment that clearly effects survival in patients with chronic obstructive lung disease. PMID- 8430397 TI - [Treatment of ventricular tachyarrhythmias with an implantable pacemaker cardioverter-defibrillator]. AB - During the period from November 1989 to January 1992, 17 patients with ventricular tachyarrhythmias received an implantable pacemaker-cardioverter defibrillator. The material consists of three female and 14 male patients with an age range of 13-66 years, mean 50.8 +/- 16.6 years. 13 patients had coronary artery disease, one cardiomyopathy, one a myocarditis sequela and two primary rhythm disorder. Four patients received epicardial and 13 endocardial electrode systems. The observation period varied from 4-30 months (april 1992), mean 15.2 +/- 8.8 months. 11 out of 17 patients (65%) experienced one or more episodes of tachyarrhythmias which was treated successfully with overdrive pacing (ramp or burst), cardioversion or defibrillation. One patient died of heart failure after an observation period of 13 months. His pacemaker-cardioverter-defibrillator had been activated more than 100 times. Two children, 13 and 15 years, were treated successfully for ventricular fibrillation four and five months after implantation of the device. The actual one year survival is 100%. Assuming that therapy with a device had not taken place, and that the six patients who experienced episodes of ventricular fibrillation died, the hypothetical probability of survival would have been 62.1 +/- 12.3%. PMID- 8430398 TI - [Anticoagulation therapy in general practice--a practice study from the health region 4]. AB - Organization of anticoagulation therapy in general practice is described on the basis of data from 56 general practitioners interviewed by questionnaire. The average number of patients controlled by each practitioner was 12.0, and 52% of the general practitioners performed the analyses themselves. In municipalities with a local hospital only 13% of the general practitioners performed the analyses themselves, compared with 75% in municipalities without a hospital. The average weekly number of Thrombotests performed by each practitioner was 12.2. When an external laboratory was used 88% were contacted earlier in the case of International Normalized Ratio (INR) > 4.8 while this was uncommon (12%) in the case of International Normalized Ratio < 2.0. We discuss the advantages and possible disadvantages of near testing of patients on oral anticoagulation therapy. PMID- 8430399 TI - [Postmenopausal hormone replacement and cardiovascular disease]. AB - We review the evidence that hormone replacement therapy protects against the development of cardiovascular disease in women. Most studies show that this treatment reduces risk of cardiovascular disease by about 50%. The results are reasonably consistent, and biologically plausible. The protective effects are mediated through effects on lipid metabolism, but also through direct effects on vessel wall physiology and blood flow. The beneficial effect of hormone replacement therapy seems to be most marked in high-risk patients and in women with severe coronary artery disease. PMID- 8430400 TI - [Minitracheotomy]. AB - Minitracheotomy is used most often in intensive care units for tracheobronchial suctioning, especially after major abdominal and thoracic surgery. In addition, minitracheotomy is used in emergency situations involving obstruction of upper airways. Percutaneous minitracheotomy means a coniotomy that should be performed under safe conditions. The Seldinger technique may facilitate insertion of the tube. The position of the tube has to be checked by aspiration of air. We describe a case with bleeding and intralaryngeal haematoma. Usually, however, minitracheotomy seems to be a valuable and safe therapeutic asset provided that the necessary precautions are taken. PMID- 8430401 TI - [Photochemotherapy--psoralen + UV-A (PUVA)--increased risk of cancer?]. AB - Photochemotherapy with psoralens + ultraviolet (UV)-A light (PUVA) has been used for about 16 years to treat psoriasis. Some reports have indicated a higher frequency of cutaneous squamous cell carcinoma among treated patients. The results are not consistent, however, and are not generally conceded. Our material from Ulleval Hospital comprises 585 patients who have been treated with PUVA from 1977 until 1991. Three patients developed cutaneous squamous cell carcinoma, but in one of them the tumour developed before start of PUVA treatment. The two others had received only very small doses. One of them had an intraepithelial cancer of the penis. In addition we registered various internal cancers in 25 patients. The Norwegian Cancer Registry compared our patient materiel with a matched control group as regards sex, age and age-specific incidence rate. There was no statistically significant difference in incidence between the patient and the control group. PMID- 8430402 TI - [Alcohol education among junior high school students. Results from a WHO educational program]. AB - A school-based social influences approach to alcohol education was tested among Norwegian 8th grade lower secondary school students. The goal of the programme was to delay onset and minimize involvement in use of alcohol among the participants. 15 schools were randomly assigned to peer-led education, teacher led education or a control condition. The programme focused on the social and environmental influences to drink alcohol, and skills to resist those influences. It consisted of five lessons over two months. Baseline and post-test data measured alcohol-use, knowledge, attitudes, skills, friends' drinking, and intentions to drink alcohol in the future. Data were collected immediately prior to and following the educational programme. The data indicate that peer-led education appears to be efficacious in reducing alcohol use and intention to use alcohol. There was no intervention effect of the peer-led programme for knowledge, attitudes or skills. There was no intervention effect for the teacher led education. PMID- 8430403 TI - [Complement analysis tests]. AB - Although routine analysis of the complement system is rarely demanded, it is important from a diagnostical and therapeutical point of view to include the relevant tests when clinically required. Complement analysis should be performed when hereditary angioedema (C1 inhibitor deficiency) is suspected, and as part of investigation of patients with immunodeficiency (recurrent infections in childhood). Furthermore, complement analysis may be required in cases of recurrent systemic neisserial infections at any age, and to follow up certain autoimmune diseases. Only a few tests are available for routine purposes. These are reviewed briefly, with emphasis on collection and storage of samples prior to analysis. It is important that these guidelines be followed, to avoid misinterpretation of the results. PMID- 8430404 TI - [How much is the decrease in blood pressure shown by repeated measurements during the same examination?]. AB - The recommendations for blood pressure determination by sphygmomanometer encourage repeated measurements. In 430 subjects aged 56-87 years, three recordings were obtained at one minute intervals. On average, the blood pressure decreased 10/3 mm Hg from the first to the third recording (p < 0.001). The decrease was more pronounced in hypertensives (12/4 mmHg) than in normotensives (7/2 mm Hg) (p < 0.01), but was little influenced by sex, age, or body mass. A change of 10/3 mm Hg is relevant with respect to the risk of cardiovascular disease, diagnosis of hypertension, and antihypertensive therapy. Also, repeated measurements may reduce the number of falsely positive hypertensives, and is recommended as the standard procedure in conventional blood pressure determination. PMID- 8430405 TI - [Air pollution, eczema and asthma in Sor-Varanger. Is the high consumption of corticosteroid ointments caused by increased tendency for eczema?]. AB - Many investigations suggest that a high degree of air-borne pollution increases the prevalence of diseases like asthma and eczema. Sor-Varanger Municipality in Finnmark County in the north of Norway receives much air-borne pollution from domestic industry and from the metallurgic industry on the Kola peninsula in North-Western Russia. We have investigated indirect parameters of morbidity caused by asthma and eczema by analyzing data on drug consumption and hospital admissions. In Sor-Varanger there is high consumption of corticosteroids for dermatological use. Consumption of anti-asthmatic drugs and number of admissions to hospital for asthma and eczema were no higher than expected. We suspect that air-borne pollution, particularly the heavy metal nickel, increases the prevalence and perhaps worsens the degree of eczema in Sor-Varanger. PMID- 8430406 TI - [Vaccination coverage in local and national perspective. Vaccination coverage before and after introduction of a local computerized system in the municipality of Sandefjord]. AB - The author presents the results from an examination in Sandefjord three years before and four years after implementation of a computer-based information system in 1986. The targets for immunization rates are well defined, but the methods for monitoring and calculation of the rates are not satisfactory. The rates seem to be reliable when given for the whole country. The fallacies caused by denominator problems when the rates are calculated for municipalities are discussed. An analysis of immunization rates and reasons for incomplete vaccination in the case of children born during the period 1983 to 1989 has been carried out in Sandefjord, an urban municipality with approximately 36,000 inhabitants. The average immunization coverage against diphtheria, tetanus, polio, whooping cough, measles, mumps and rubella (for children aged 2-5 years) showed record levels, i.e. about 97-98% of children for the whole period. The predominant reason for incomplete vaccination was that the parents did not want their child to be exposed to vaccination. In the case of a very few children the vaccination programme was delayed by medical contradictions. PMID- 8430407 TI - [Euthanasia and choice of drugs]. PMID- 8430408 TI - [Violent behavior of patients--can it be predicted?]. PMID- 8430409 TI - [Allergy]. PMID- 8430410 TI - [Physicians' practice in declaring the patients fit]. PMID- 8430411 TI - [Data--an intricate tool in the hands of insurance authorities?]. PMID- 8430412 TI - [The right to die with dignity--reflections from a Christian point of view]. PMID- 8430413 TI - [Current environmental situation--physician's responsibility and response]. PMID- 8430414 TI - [Central prioritization of health services--do the authorities create A and B patients?]. PMID- 8430415 TI - [A framework plan for distribution of services between hospitals in Oslo]. PMID- 8430416 TI - [Music and hearing damage. If it's too loud, you are to old!]. PMID- 8430417 TI - Alteration of tissue and serum sphinganine to sphingosine ratio: an early biomarker of exposure to fumonisin-containing feeds in pigs. AB - Fumonisins are a group of naturally occurring compounds produced by the fungus Fusarium moniliforme. They are believed to be the etiologic agent of several animal diseases associated with consumption of corn-based feeds including porcine pulmonary edema. Recently it was shown in vitro that fumonisins are specific inhibitors of sphingosine and sphinganine N-acyltransferases. Inhibition of these enzymes in cultured cells results in the accumulation of free long chain sphingoid bases, specifically sphingosine and sphinganine, and the depletion of complex sphingolipids. In this study, tissues and serum from male SPF pigs fed a nutritionally balanced diet containing corn or corn screenings naturally contaminated with fumonisins for up to 14 days were analyzed for free sphingoid bases and complex sphingolipids. Total fumonisins (B1 and B2) in the diets were analyzed at 0 (< 1), 5, 23, 39, 101, and 175 ppm. Pulmonary edema only occurred at 175 ppm, while histologic liver damage was present at > or = 23 ppm, and serum liver enzymes were significantly elevated at > or = 101 ppm. The results of this study show that free sphinganine is elevated in liver, lung, and kidney, from pigs consuming feeds containing fumonisins at total fumonisin concentrations of 23 ppm or greater. Sphingosine is also elevated in a dose-dependent manner, but to a lesser extent than sphinganine. The consequence of this differential inhibition is that the ratio of sphinganine to sphingosine increases, suggesting that sphinganine N-acyltransferase is the preferred target for fumonisins. Elevation of free sphinganine and free sphingosine in serum paralleled the increases in tissues. Statistically significant increases in the ratio were observed at feed concentrations as low as 5 ppm total fumonisins and in pigs (at higher concentrations) in which other serum biochemistry parameters and tissue morphology were not altered. Elevated ratios were also observed in serum from pigs fed pure fumonisin B1. The sensitivity of the ratio indicates that it could serve as an effective biomarker for consumption of fumonisin-containing feeds. In addition, the data supports the hypothesis that inhibition of sphingosine and sphinganine N-acyltransferase plays an important role in the pathogenesis of animal diseases associated with consumption of feed containing fumonisins. PMID- 8430418 TI - Effect of progesterone pretreatment on cadmium toxicity in the male Fischer (F344/NCr) rat. AB - A previous report has indicated that progesterone pretreatment can markedly reduce cadmium toxicity in male NAW mice. Therefore we examined the effects of progesterone pretreatment on cadmium toxicity in male Fischer (F344/NCr) rats. A single sc injection of 20 mumol CdCl2/kg proved nonlethal over 24 hr but caused the typical spectrum of testicular lesions in these rats. However, when rats were pretreated with progesterone (100 mg/kg, sc, -48, -24, and 0 hr) and then given cadmium (20 mumol CdCl2/kg, 0 hr), this dose of cadmium proved very toxic, unexpectedly causing a 53% mortality. Progesterone pretreatment had no effect on cadmium-induced testicular lesions in surviving rats. Significant elevations in serum lactate dehydrogenase (LDH) activity, indicative of hepatotoxicity, were also observed in progesterone-pretreated rats given cadmium as compared to rats given cadmium alone. Progesterone pretreatment had no effect on the distribution of cadmium to liver, kidney, or testes. Progesterone pretreatment also had no effect on the cadmium-induced increases in hepatic or renal metallothionein (MT) or hepatic or testicular MT mRNA levels. In contrast, levels of the testicular cadmium-binding protein (TCBP) in progesterone-pretreated rats were doubled. These results indicate that, contrary to previously reported data for the mouse, progesterone pretreatment increased the lethality of cadmium in male Fischer (F344/NCr) rats and had no effect on cadmium-induced testicular toxicity. The mechanism by which progesterone enhanced cadmium toxicity, especially cadmium induced hepatotoxicity, deserves further study. PMID- 8430419 TI - Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the epidermal growth factor receptor in hepatic plasma membranes of rainbow trout. AB - Time- and dose-dependent alterations in epidermal growth factor receptor (EGF-R) ligand binding and protein kinase activity were observed in hepatic plasma membranes of TCDD-treated rainbow trout. Trout were dosed by a single ip injection of TCDD in a corn oil vehicle. A single ip injection of TCDD (10 micrograms TCDD/kg body weight) caused a maximal reduction of EGF binding to hepatic plasma membranes by 10 days posttreatment which remained reduced until Day 40. EROD activity in the liver microsomes of TCDD-treated trout increased relative to untreated fish over the course of the study. Protein kinase C and tyrosine kinase activity as well as EGF receptor phosphorylation was greater in livers of treated than in those of control fish within 5 days but returned to control levels by 40 days postinjection. In a dose-response study, EGF binding was reduced in a dose-dependent manner with an ED50 of 0.17 micrograms TCDD/kg wet weight while EROD activity was induced with an ED50 of 0.79 micrograms TCDD/kg. The reduction in EGF binding was correlated to an increase in EROD activity, protein kinase C activity, and tyrosine kinase activity but was negatively correlated to EGF receptor phosphorylation. Of the parameters examined in both the time course and dose studies, protein kinase C was the best predictor of the reduction of EGF binding to hepatic plasma membranes of rainbow trout. The results from this study are consistent with the hypothesis that the mode of action of TCDD on the EGF receptor is in part mediated through the protein kinase C activity. It also suggests that the toxic mode of action of TCDD is similar in rainbow trout and mammals. PMID- 8430420 TI - Alterations in biodistribution of cocaine may explain differential toxicity in pregnant and postpartum rats. AB - Biodistribution of cocaine in female rats before and after delivery of litters was studied in order to provide an explanation for the observed differences in cocaine-induced toxicity in pregnant and postpartum rats. Cocaine (20 mg/kg, ip) given to female rats at 16 days of gestation or 20 days of gestation did not have any apparent neurotoxic effects in the mothers. Similarly, no apparent toxicity was observed in virgin animals injected with the same dosage of cocaine. However, administration of the same dosage of cocaine to rats after delivery caused clonic tonic convulsions and death within 5 min of injection. In an effort to explain these differences, the biodistribution of cocaine was investigated by injecting levo-[benzoyl-3,4-3H(N)] cocaine (20 mg/20 microCi/kg, ip) to nonpregnant rats and rats at different gestation periods and after delivery. The mean serum/brain ratios (concentration of cocaine in serum divided by cocaine concentration in the brain) of cocaine 5 min after injection in rats at 16 days gestation, 20 days gestation, within a 12-hr period after delivery, and in control virgin animals were 2099, 1132, 427, and 632, respectively. These results indicate that the biodistribution of cocaine is altered at parturition. This may explain the increased central nervous system toxicity in rats treated with cocaine after delivery. Results from this investigation may be useful in evaluating complications during labor and delivery which have been associated with cocaine abuse by pregnant women. PMID- 8430421 TI - Mutagenicity of hydralazine in mammalian cells and bacteria. AB - The genotoxicity of hydralazine (HDZ), an antihypertensive agent, was evaluated in mammalian cells and bacteria. The formation of mutants at the hypoxanthine guanine phosphoribosyl transferase locus in an adult rat liver cell line ARL 18 was determined. Bacterial mutagenicity was assessed in Salmonella typhimurium strains TA100 and TA102. The latter strain was chosen because it has A:T bases at the reversion site and HDZ has been reported to interact with thymidine. HDZ was mutagenic to ARL 18 cells with a concentration-dependent increase in mutants observed at 5 x 10(-6) to 5 x 10(-4) M. At 5 x 10(-4) M HDZ, there were 110 mutants/10(6) colony-forming cells compared to 129 for cells exposed to 10(-4) M benzo(a)pyrene, a known genotoxin. Bacterial mutants were observed with HDZ in both strains in the absence of an activating system. HDZ also induced mutants in the presence of S-9 from Aroclor-induced rat liver or uninduced rabbit liver. These results were consistent with previous reports of the mutagenicity of HDZ in TA100 and extend the observations to TA102, a strain designed to detect oxidative damage. This study also provides the first evidence of the mutagenicity of HDZ in mammalian cells. These data support the genotoxicity of HDZ in in vitro systems. PMID- 8430422 TI - Physiologically based models for bone-seeking elements. IV. Kinetics of lead disposition in humans. AB - A model of lead disposition in humans has been developed. In addition to liver, kidney, other well-perfused and poorly perfused tissues, the model incorporates bone as an explicit compartment whose volume, composition, and metabolic activity are all age-dependent. The rates of all transfers of lead out of the blood are related to plasma lead. The relationship between plasma lead and blood lead is expressed as a capacity-limited binding associated with the erythrocyte. Interchanges of lead between plasma and bone are modeled as partially interactive events linked in series. Blood traversing the bone passes first through a surface region of rapid exchange before entering the metabolically active region of bone, where lead is incorporated with calcium into mineralizing bone and returned to the blood with resorbing bone. After leaving the larger vessels, plasma superfusate enters the canalicules that feed bulk bone, where lead diffuses radially outward into the total bone volume. Model simulations are compared with data from epidemiologic and experimental dietary and inhalation studies in adults. PMID- 8430423 TI - Reduced methylmercury accumulation in a methylmercury-resistant rat pheochromocytoma PC12 cell line. AB - Rat pheochromocytoma PC12 cell sublines with increasing resistance to methylmercury (MeHg) were isolated by exposure to MeHg in two sequential steps. The strongest resistance (PC12/TM) among the clones obtained by the first cloning was characterized. PC12/TM cells exhibited about an 8- to 10-fold increase in resistance compared with parent PC12 cells on the bases of the concentration required for 50% inhibition of growth and colony-forming activity. PC12/TM cells accumulated smaller amounts of MeHg (one-half to one-fifth) than parent PC12 cells. This reduced MeHg accumulation in PC12/TM cells resulted from the slow uptake and rapid efflux. A close correlation between reduced MeHg accumulation and MeHg resistance was found among seven sublines of PC12 cells with different sensitivity to MeHg. The reduced retention of MeHg in PC12 sublines was also well correlated with the sensitivity to MeHg. Phenol-3,6-dibromophthalein disulfonate inhibited MeHg efflux from PC12/TM cells and increased its accumulation. These results suggest that efflux of MeHg from PC12/TM cells is associated with glutathione. PMID- 8430424 TI - Cadmium-induced attenuation of coronary blood flow in the perfused rat heart. AB - The present studies were undertaken to test the hypothesis that cadmium alters myocardial metabolism, producing effects on coronary blood flow. Hearts isolated from adult, male Sprague-Dawley rats were control-equilibrated and then treated with Cd (0.1-100 microM). Myocardial O2 consumption (MVO2), lactate production, contractile activity, and coronary flow were determined at regular intervals. Parallel experiments were also performed in hearts administered 0.5 mM KCN to compare the effects of cadmium on myocardial lactate release with that of a known inhibitor of mitochondrial respiration. Coronary flow decreased significantly in response to Cd. This effect was dose-related (ED50 = 0.4 microM Cd), achieving maximal levels within the initial 5 min of exposure and persisting thereafter. Cd treatment also significantly decreased myocardial contractile activity [ED50 = 2.4 microM Cd (+dP/dt)] and MVO2 in a dose-dependent manner. This latter effect was time-dependent with an ED50 value of 2.1 microM at 5 min, as compared to a value of 0.5 microM Cd at 30 min. Lactate levels measured in the coronary effluent were unaffected by Cd treatment, except at the highest dose. Collectively, the results of this study failed to support a metabolic mechanism as the basis for the observed changes in coronary flow in response to cadmium administration. Instead, these results suggest an alternative hypothesis that Cd disturbs coronary flow via a mechanism involving direct actions on the coronary vasculature. PMID- 8430425 TI - Effect of vitamin B12 status on selenium methylation and toxicity in rats: in vivo and in vitro studies. AB - Animals are known to convert inorganic selenium to less toxic methylated compounds such as dimethylselenide (DMSe) and trimethylselenonium (TMSe). This study investigated the role of vitamin B12, a cofactor of methionine synthetase, in selenium methylation in the rat. Vitamin B12-depleted rats expired 16% of dosed 75Se-selenite as DMSe compared to 45% for control rats and excreted less TMSe in the urine (6.1% of dose) than control (9% of dose) rats. At the same time, higher (p < 0.05) tissue (liver, kidney, muscle) selenium levels and lower (p < 0.05) blood selenium levels were found in vitamin B12-deficient rats. Primary hepatocytes from vitamin B12-deficient rats volatilized 15% of selenite in incubation medium in 5 hr as compared to 49% in hepatocytes from control rats. Hepatocytes from vitamin B12-deficient rats were less resistant to selenite toxicity. In vitro methylation of selenium with liver extract from vitamin B12 deficient rats showed one-third to one-half the rate of volatilization of selenium as compared to control rats. S-adenosylmethionine was required for this reaction. These results show that vitamin B12 deficiency significantly decreases the ability of rats to methylate selenium. PMID- 8430426 TI - Reduction of azo dyes during in vitro percutaneous absorption. AB - The azoreduction of phenylazo-2-naphthol (Sudan I), 5-(phenylazo)-6 hydroxynaphthalene-2-sulfonic acid [aniline subsidiary color of FD&C Yellow No. 6 (ANSC)], and phenylazophenol [Solvent Yellow 7 (SY7)] in skin during percutaneous absorption was measured and the contributions of cytosolic and microsomal reductions were characterized. By using a series of azo dyes with a common U-14C labeled phenylazo moiety, percutaneous absorption and metabolism were measured in vitro in flow-through diffusion cells with Sencar mouse, hairless guinea pig, and human skin. Azoreductase assays using subcellular fractions from skin of all species were used to examine intracellular rates and distribution for the series of dyes. The absorption of the lipophilic dyes Sudan I and SY7 from a finite surface dose of 5 micrograms/cm2 was extensive in all species. In mouse, 32.8 +/- 2.8% of the applied Sudan I dose and 64.1 +/- 3.3% of the applied SY7 dose were absorbed in 24 hr. In the hairless guinea pig, 57.6 +/- 5.9% of the applied Sudan I dose and 67.8 +/- 4.6% of the applied SY7 dose were absorbed in 24 hr. Human skin was least permeable, with 26.4 +/- 6.7% of the applied Sudan I dose and 36.1 +/- 4.5% of the applied SY7 dose absorbed in 24 hr. Sudan I and SY7 were extensively reduced in skin of all species during percutaneous absorption (29.5 and 26.5%, respectively, of the absorbed dose in human skin and greater than 50% of the applied dose in other species). ANSC was the least absorbed, with 5% or less penetrating. SY7 was preferentially reduced in the skin cytosol of all species, whereas Sudan I was equally reduced in the skin cytosol and microsomal fractions. The site of azoreduction in the cell may affect the metabolic fate of the liberated arylamine. The extensive azoreduction observed during percutaneous absorption may modulate the toxicities of these compounds and must be considered when effective doses are determined for quantitative risk assessments from dermal exposures. PMID- 8430427 TI - Perfluorodecanoic acid noncompetitively inhibits the peroxisomal enzymes enoyl CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase. AB - The mechanisms of the inhibition of hepatic peroxisomal beta-oxidation by the peroxisome proliferator PFDA3 were studied. Female Sprague Dawley rats were given a single ip injection of either 0, 10, or 40 mg/kg PFDA or were placed on a diet supplemented with the peroxisome proliferator ciprofibrate (0.01%). After 2 weeks, the rats were killed, and hepatic peroxisomes were isolated by discontinuous sucrose gradient centrifugation. Treatment of rats with either PFDA or ciprofibrate increased the individual activities of each of the enzymes in the peroxisomal beta-oxidation pathway. Similarly, treatment of rats with ciprofibrate greatly increased total peroxisomal beta-oxidation, but peroxisomal beta-oxidation was slightly decreased in rats treated with 40 mg/kg PFDA. In vitro inhibition studies found that PFDA was a noncompetitive and reversible inhibitor of both the activities of the peroxisomal bifunctional protein, namely enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase. The Ki of the inhibition was approximately 5 microM. PFDA only slightly inhibited the activity of peroxisomal fatty acyl CoA oxidase, and did not inhibit peroxisomal thiolase activity. We therefore conclude that PFDA inhibits peroxisomal beta-oxidation by noncompetitively inhibiting the peroxisomal bifunctional enzyme. PMID- 8430428 TI - Stable and inducible arsenite resistance in Chinese hamster cells. AB - A number of Chinese hamster V79 cell sublines that are arsenite resistant or arsenite sensitive were isolated. After more than 6 months of growth in the absence of arsenite, these sublines still maintain their arsenite-resistant or sensitive phenotypes. At least some arsenite-resistant cell lines are also cross resistant to sodium arsenate and potassium antimonyl tartrate. Wild-type or arsenite-resistant sublines show a further increase in resistance to toxic concentrations of arsenite after pretreatment with a nontoxic concentration. Pretreatment of cells with a nontoxic dose of arsenite also increased their survival to toxic doses of antimonite. Likewise, pretreatment with antimonite increased survival to arsenite as well as to antimonite. The inducible arsenite resistance increases with pretreatment time, reaching a plateau after 8 hr of pretreatment. Fusion of arsenite-resistant cells with arsenite-sensitive cells demonstrated a clear dominance of arsenite resistance. These results suggest that mammalian cells contain an arsenite/antimonite pump whose activity may be modulated by prior exposure to arsenite or antimonite. PMID- 8430429 TI - COR pulmonale is caused by monocrotaline and dehydromonocrotaline, but not by glutathione or cysteine conjugates of dihydropyrrolizine. AB - Monocrotaline (MCT) produces pulmonary hypertension and right ventricular hypertrophy in rats. It is generally believed that MCT must undergo hepatic metabolism to reactive metabolites that are subsequently transported to the lungs to induce a pneumotoxic response. Several studies suggest that dehydromonocrotaline (MCTP) is the reactive intermediate that initiates pulmonary toxicity. We recently identified two other MCT metabolites, the glutathione and N acetylcysteine conjugates of 6,7-dihydro-7-hydroxy-1-hydromethyl-5H-pyrrolizine (DHP). To determine the potential pulmonary toxicity of the glutathione conjugate (DHP-GSH) and the unacetylated cysteine conjugate precursor (DHP-Cys) of the N acetylated excretion product, we conducted parallel in vivo toxicity studies with DHP-GSH, DHP-Cys, MCT, and MCTP. Relative pneumotoxicity was evaluated by measurements of right ventricular pressure (RVP), ventricular weight ratio (RV/LV+S), subjective histopathology, and measurements of components of the arteriolar wall. Animals given a single injection of MCT (60 mg/kg) developed pulmonary hypertension at the end of 3 weeks, as indicated by significant elevation in RVP when compared to the controls (22.1 +/- 2.4 mm Hg vs 13.2 +/- 0.8 mm Hg). A parallel and significant increase in RV/LV+S was also evident: 0.37 +/- 0.021 (MCT) vs 0.299 +/- 0.011 (control). Histopathology showed marked alterations in both pulmonary vasculature and parenchyma in MCT- and MCTP-treated animals. MCTP (1 mg/kg) caused a significantly elevated RVP (MCTP vs control: 28.1 +/- 3.4 mm Hg vs 16.8 +/- 0.97 mm Hg) and an increased RV/LV+S (MCTP vs control: 0.445 +/- 0.051 vs 0.284 +/- 0.026). Both MCT- and MCTP-treated rats had increased arteriolar medial thickness and decreased lumen diameter, but MCTP treated rats had a milder vascular inflammatory response and less parenchymal lesions. Neither DHP-GSH (24 or 12 mg/kg) nor DHP-Cys (12 mg/kg) caused detectable changes in pulmonary circulation and no structural alteration in the lung was observed in these treatment groups. Although they are all pyrrolic metabolites of MCT, these studies demonstrate that only MCTP but not the glutathione or cysteine conjugates, is pneumotoxic at the doses tested. PMID- 8430430 TI - Enhanced hepatic expression of P450IIE1 following inhalation exposure to pyridine. AB - Previous research has shown that intraperitoneal injection of the solvent pyridine results in a dose- and time-dependent induction of hepatic P450IIE1 in rats, with a twofold increase observed at 6 hr post-treatment. However, inhalation is the primary route of exposure expected for humans exposed to pyridine in laboratory and industrial settings or in tobacco smoke. In view of the potency and rapidity of induction associated with pyridine treatment, research was initiated to examine whether inhalation of the solvent at the current threshold limit value level of 5 ppm or at an elevated level of 440 ppm resulted in sufficient nasal and pulmonary uptake and systemic redistribution to cause increased hepatic expression of rat cytochrome P450IIE1. Rats were exposed, 6 hr/day, to 5 ppm pyridine for 4 days; to 440 ppm pyridine for 1 or 4 days; or to filtered air for 1 or 4 days. Rats were sacrificed 18 hr after the last exposure. Liver samples were taken from the air- and pyridine-exposed rats. Expression of immunoreactive P450IIE1 was examined in tissue sections using immunohistochemistry and in microsomal preparations using Western blot analyses. Hepatic tissue obtained from rats exposed to 5 ppm pyridine for 4 days or to 440 ppm pyridine for 1 or 4 days displayed significantly elevated levels of immunoreactive P450IIE1. Immunohistochemical analyses of liver sections showed that inhalation of pyridine at 5 ppm resulted in an elevated expression of P450IIE1 in hepatocytes surrounding terminal hepatic venules (THVs). Exposure to 440 ppm pyridine caused both increased expression of IIE1 in hepatocytes surrounding THVs after a single 6-hr exposure and an approximately three- to fourfold increase in the total number of cells expressing IIE1 after 4 days of exposure. Dilution analysis of immunohistochemically stained tissue revealed a significant difference between air-exposed controls and pyridine-exposed rats, at all exposure levels. P450IIE1 levels in microsomes isolated from rats receiving 5 and 440 ppm pyridine for 4 days were increased 2.5- and 10-fold as judged from scanning laser densitometry. This study reveals that inhalation of pyridine vapors at levels equal to or greater than the current threshold limit value of 5 ppm results in significant hepatic induction of P450IIE1 in rats. PMID- 8430431 TI - Enhancement of renal and hepatic glutathione metabolism by dimethylthiourea. AB - Dimethylthiourea (DMTU), a free radical scavenger, was administered to rats to study its effect on renal and hepatic glutathione metabolism, since it is a potential sulfhydryl donor. Six hours following DMTU, renal GSH content was significantly (P < 0.05) increased (10%), and was increased further after 24 h (28%) (P < 0.001). Hepatic GSH content was also significantly (P < 0.001) elevated at 6 and 24 h (5 and 33%, respectively). Seven days of daily DMTU therapy significantly (P < 0.001) increased renal and hepatic GSH content by 36 and 54%, respectively, which was associated with a significant (P < 0.001) increase in the renal activities of glutathione peroxidase (GP) by 38%, glutathione transferase (GT) by 92%, and glutathione reductase (GR) by 19% (P < 0.05). Significantly increased activities of hepatic GP by 84% (P < 0.01) and GT by 101% (P < 0.001) also occurred in DMTU-treated rats after 7 days of continuous therapy. From these data, we conclude that DMTU stimulates renal and hepatic GSH metabolism, which may be important in mediating DMTU's protective effect against free radical-induced tissue injury. PMID- 8430432 TI - The toxicological effect of cyclophosphamide on acetylcholinesterase activity. AB - Cyclophosphamide (CP; 2.5-10.0 mM) reversibly inhibits acetylcholinesterase (AChE) activity of chicken brain (58-84%) in a concentration-dependent manner, the IC50 (the concentration causing 50% inhibition) being about 2.0 mM. Lineweaver-Burk plots indicated that the nature of this inhibition is mixed; intercept and slope replots are straight lines, showing that the inhibition is complete rather than partial. The value of Km for AChE is found to be 108 microM, while values of KI and Ki are found to be 1.5 and 5.1 mM, respectively, Ki is greater than KI indicating that uncompetitive inhibition is predominant over noncompetitive inhibition. PMID- 8430433 TI - Differential aluminium lactate toxicity in rabbits using either aqueous solutions or liposomal suspensions. AB - The present paper deals with the toxic effects of aluminum lactate (Al(lact)3) in rabbits. Experimental animals were injected with 6.2 mg of Al(III) as Al(lact)3 aqueous solutions at neutral pH for 21 days. Histological examination showed different pathological lesions to the myocardial tissue, spleen, kidney and liver, with no relevant effects to lungs and CNS. On the contrary, rabbits injected with 60 micrograms of Al(III) as Al(lact)3 in a liposome suspension for 42 days showed a large infarctual zone in the spinal cord with the metal accumulation in the necrobiotic neurons. Pharmacological implications of these findings are also discussed. PMID- 8430434 TI - Effect of exposure route on measurement of blood pressure by tail cuff in F-344 rats exposed to OTTO Fuel II. AB - Male Fischer-344 rats demonstrated a dose-response of blood pressure (BP) to increasing doses of propylene glycol dinitrate (PGDN), the major constituent of OTTO Fuel II (OFII) following administration by subcutaneous injection. Dermal application of the same doses to separate groups of rats resulted in variable responses of BP that were unrelated to dose. A nose-only exposure system was developed but no effect on BP was observed in rats exposed to a nearly saturated atmosphere of PGDN (approx. 750 mg/m3 at 25 degrees C). This study has indicated both the difficulties associated with the use of tail cuff measurement of BP and the need for either a more sensitive or more specific biomarker of effect for exposure to nitrate esters. PMID- 8430435 TI - Semicarbazide protection from in vivo oxidant injury of vascular tissue by allylamine. AB - Allylamine is a specific cardiovascular toxin that causes vascular and myocardial lesions. Previous studies showed that allylamine-induced chronic lesions are markedly reduced by semicarbazide, an inhibitor of semicarbazide-sensitive amine oxidase (SSAO), and that allylamine is metabolized to the aldehyde, acrolein, by SSAO. We hypothesized that inhibitors of SSAO might reduce the acute cardiovascular toxicity of allylamine. To test our hypothesis, we fed 150 mg/kg allylamine to semicarbazide-pretreated (3 h; 98 mg/kg) rats. Animals were sacrificed 1 h after allylamine treatment. Aorta, epicardium, and endocardium were assayed for SSAO, glutathione peroxidase, catalase, thiol status and lipid peroxidation. SSAO activity was decreased significantly in aorta, epicardium and endocardium. Activity was 30-times higher in aorta than in epicardium and endocardium. A striking decrease in malonaldehyde level (lipid peroxidation) was found in aorta of pretreated rats as compared to allylamine-only treated rats. The reduction of free-SH content in aortic mitochondria was also attenuated in pretreated rats. Changes were not so marked in epicardium and endocardium. These results suggest that in vivo pretreatment with semicarbazide at least partially protects aortic mitochondria from allylamine toxicity. The mechanism can be explained on the basis of the fact that semicarbazide inhibits acrolein formation in allylamine-treated rats. PMID- 8430436 TI - Analysis of mammalian metallothionein isoforms by high-resolution SDS-gel electrophoresis. AB - Metallothioneins (MTs) are cysteine-rich heavy metal-binding proteins, whose possible functions are thought to be the protection against toxic metals as well as the regulation of essential metals. It is known that there are several MT isoforms, but the biological roles of the individual isoforms have not been elucidated. To facilitate the functional analysis of these isoforms, we improved an analytical method of MTs developed previously, which is based on a denaturing gel electrophoresis of chemically modified MTs. The established technique makes it possible not only to separate MT isoforms with a high resolution, but to estimate the levels of the individual isoforms by analyzing directly crude cell extracts. By this method, six MT isoforms were identified in the extracts of Cd exposed human cells. It was also revealed that there is an apparent heterogeneity of the rat liver MT; five isoforms were identified in the liver extracts of Cd injected rats. The present method will be useful in the functional analysis of the MT isoforms, as well as in a variety of aspects of the MT studies. PMID- 8430437 TI - Steroidal alkaloid toxicity to fish embryos. AB - Embryos of two species of fish were evaluated for their suitability as model systems for steroidal alkaloid toxicity, the Japanese rice fish, medaka (Oryzius latipes) and the rainbow trout (Oncorhynchus mykiss). Additionally, the equine neurotoxic sesquiterpene lactone repin, was also tested. A PROBIT program was used to evaluate the EC1, EC50 and EC99 as well as the associated confidence limits. The steroidal alkaloids tested were the Solanum potato glycoalkaloids alpha-chaconine, alpha-solanine, the aglyclones solanidine and solasodine and the Veratrum alkaloid, jervine. Embryo mortality, likely due to structural or functional abnormalities in the early development stages of the embryo, were the only response observed in both species. The rainbow trout exhibited a toxic response to chaconine, solasidine, repin and solanine but the medaka embryos were only affected by the compounds, chaconine and solanine. Rainbow trout may indeed serve as a good lower vertebrate model for studying the toxicity of steroidal alkaloids. PMID- 8430438 TI - In vitro assessment of teratogenic potential of organotin compounds using rat embryo limb bud cell cultures. AB - Assessment of the relative teratogenic potential of bis(tri-n-butyltin)oxide (TBTO), tri-n-butyltin chloride (TBT), and its metabolites, i.e., (3 OH)hydroxybutyl dibutyltin chloride ((3-OH-Bu)DBT), di-n-butyltin dichloride (DBT), and butyltin trichloride (MBT) have been conducted using rat embryo limb bud cell cultures (LBC) to gain some knowledge of TBT embryotoxicity and DBT teratogenicity. Triphenyltin chloride (TPT), trimethyltin chloride (TMT), and triethyltin bromide (TET) have also been tested to obtain data for validation of LBC as a teratogen prescreening for organotin compounds. Fifty percent inhibition concentration for cell proliferation (IP50), and for cell differentiation (ID50), and the ratio of the former to the latter (P/D ratio) were obtained. The ID50 values in increasing order were as follows; TPT, DBT < TBT, TBTO < (3-OH-Bu)DBT < TET < TMT << MBT. With the exception of MBT, organotin compounds tested were very strong inhibitors of cell differentiation (ID50; 0.13-1.71 microM) and cell proliferation (IP50; 0.12-2.81 microM). P/D ratios for TBT, (3-OH-Bu)DBT, DBT and MBT were 1.0, 1.43, 1.32 and 1.08, respectively. These results suggest that the proximal toxin of DBT teratogenicity is DBT itself, and TBT is rather embryocidal than teratogenic so that TBT might mask the teratogenic and/or fetotoxic effects of its metabolites. PMID- 8430439 TI - Prevention of renal toxicity of cis-diamminedichloroplatinum by dithiocarbamates in rats. AB - The protective effects of various dithiocarbamates such as N-benzyl-D-glucamine dithiocarbamate (BGD), N-p-hydroxymethylbenzyl-D-glucamine dithiocarbamate (HBGD), N-p-carboxybenzyl-D-glucamine dithiocarbamate (CBGD), and N-methyl-D glucamine dithiocarbamate (MGD) on DDP-induced renal toxicity in rats were studied. The rats received the simultaneous i.v. injection of DDP (20 mumol/kg) and a chelating agent (40 mumol/kg). Significant increase in blood urea nitrogen (BUN) level was observed 5 days after DDP injection. The increase in BUN level was completely prevented by only HBGD and CBGD among these chelating agents. Treatment with CBGD completely prevented against DDP-induced body weight loss. BGD and MGD treatment did not prevent the increase in BUN level or body weight loss. HBGD and CBGD were the most effective in decreasing the renal platinum content, resulting in maximum protection against the DDP-induced renal damage. The antitumor efficacy of DDP in the Walker 256 carcinoma-bearing rats was not affected by CBGD administration. PMID- 8430440 TI - In vivo effect of 7,8-benzoflavone on aryl hydrocarbon hydroxylase activity of mouse liver microsomes. AB - In vivo administration of 3-methylcholanthrene (MC) and 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) produced much higher hepatic microsomal aryl hydrocarbon hydroxylase (AHH) induction than 7,8-benzoflavone (7,8-BF) in both aromatic hydrocarbon (Ah)-responsive and nonresponsive strains of mice. Simultaneous treatment or pre-treatment with 7,8-BF produced an inhibitory effect on AHH induction by MC or TCDD (i.e., the degrees of the inhibition, with TCDD, were 28% in the Ah-responsive strain C57BL/6N (C57) mice and 45% in the nonresponsive strain DDD;Qdj (DDD) mice). However, posttreatment with 7,8-BF was inclined to promote the induction of AHH by MC or TCDD (i.e., the degrees of the enhancement, with MC, were 15% in C57 mice and 45% in DDD mice). These results may suggest that the inhibitory effect of 7,8-BF in vivo is limited not to the combination of AHH inducer (MC or TCDD) but to its application of timing or Ah responsiveness. PMID- 8430441 TI - 7 alpha-Methyl-17 alpha-(E-2'-[125I]iodovinyl)-19-nortestosterone: a new radioligand for the detection of androgen receptor. AB - We have synthesized two gamma-emitting, 125I-labeled steroids, E- and Z-7 alpha methyl-17 alpha-(2'-[125I]iodovinyl)-19-nortestosterone [125I](E- and Z-MIVNT) for specific labeling of androgen receptors. [125I]E- and [125I]Z-MIVNT were synthesized stereospecifically from E- and Z-7 alpha-methyl-17 alpha-(2'-tri-n butylstannyl-vinyl)-19-nortestosterone. The tin adducts were prepared by addition of tri-n-butyltin hydride to 7 alpha-methyl-17 alpha-ethynyl-19-nortestosterone, and after purification they were converted in high yield to the [125I]MIVNT isomers by reaction with 125I (generated in situ by oxidation of [125I]iodide with chloramine T). The 125I-labeled products were purified by high-performance liquid chromatography, and their mass determined with an ultraviolet detector (specific activity of both, approximately 2,200 Ci/mmol). In rat prostate cytosol, [125I]E-MIVNT bound with high affinity to a single class of binding sites. Nonspecific binding in the presence of 5 alpha-dihydrotestosterone was relatively low, and compared favorably with that obtained in parallel studies with [3H]methyltrienolone (R1881). The E-isomer bound prostate cytosol with at least twice the affinity of the Z-isomer; therefore, the interaction of the E isomer with the androgen receptor as well as other steroid receptors was studied in greater detail. Complexes of the androgen receptor with [125I]E-MIVNT as well as [3H]R1881 dissociate very slowly at 4C (kdiss for both = 0.04 h-1). Displacement studies showed that the interaction of [125I]E-MIVNT with the androgen receptor is highly specific. Competition studies showed that unlabeled E MIVNT binds poorly to other steroid receptors in rat tissue cytosols. These binding properties make [125I]E-MIVNT a promising ligand for study of the androgen receptor, and [123I]E-MIVNT a potential imaging agent for the detection of androgen-dependent tumors, such as prostate cancer. PMID- 8430442 TI - 11-Dehydrocorticosterone in the presence of carbenoxolone is a more potent sodium retainer than corticosterone. AB - In vivo, corticosterone and 11-dehydrocorticosterone are interconverted in the liver and possibly kidney by 11 beta-hydroxysteroid dehydrogenase. In an effort to evaluate the relevance of this reversible reaction in relation to urinary sodium and potassium excretion, we investigated the effects of 11 dehydrocorticosterone in the presence and absence of carbenoxolone, a potent inhibitor of the oxidative component of 11 beta-hydroxysteroid dehydrogenase, and compared them with the effects of similar doses of corticosterone in carbenoxolone-treated rats. All experiments were performed on adrenalectomized male rats. Here we describe that in carbenoxolone-treated rats 11 dehydrocorticosterone and corticosterone display antinatriuretic activity, although under the conditions of this study 11-dehydrocorticosterone is a more potent sodium retainer than its parent steroid corticosterone. In addition, the antinatriuretic effects of 11-dehydrocorticosterone (like the antinatriuretic effects of corticosterone in carbenoxolone-treated rats) were blocked by the specific antimineralocorticoid RU28318. PMID- 8430443 TI - Synthesis of new amino acid and peptide derivatives of estradiol and their binding affinities for the estrogen receptor. AB - A series of amino acid and peptide derivatives of estradiol have been synthesized by coupling 17 beta-aminoestra-1,3,5(10)-trien-3-ol, 17-hydrazonoestra-1,3,5(10) trien-3-ol with amino acids or peptides, using tetrahydrothiazole-2-thione, N hydroxy-1,4-epoxycyclohex-5-ene-2,3-dicarbonylimide, benzotriazolyloxy tris(dimethylamino)phosphonium hexafluorophosphate, and p-nitrophenol as reagents. N-protected peptidyl steroids were deprotected by traditional methods. The relative binding affinities of the deprotected derivatives to the estrogen receptor were determined by competitive radioligand binding assay. PMID- 8430444 TI - A time-dependent inactivation of aromatase by 19-substituted androst-4-ene-3,6,17 triones. AB - Diastereomeric (19S)- and (19R)-19-ethynyl-19-acetoxy derivatives of androst-4 ene-3,6,17-trione (AT) (9 and 10) and 19,19-difluoro AT (12) were synthesized. The 19,19-difluoro compound (12) was an effective competitive inhibitor of human placental aromatase with an inhibition constant (ki) of 1.8 microM but the acetylenic 9 and 10 were poor inhibitors of the enzyme with k(is) of 75 and 67 microM, respectively. Inhibitor 12 caused a time-dependent, biphasic loss of aromatase activity in the presence of reduced nicotinamide-adenine-dinucleotide phosphate (NADPH) in air, whereas the other two caused a time-dependent, pseudo first-order inactivation of the activity with rate constants for inactivation of 0.250, 0.077, and 0.065 min-1 for steroids 12, 9, and 10. NADPH was required for the time-dependent inactivation, and the substrate androst-4-ene-3,17-dione prevented it. L-Cysteine did not protect aromatase from the inactivation. PMID- 8430446 TI - Standards for bone marrow donation. PMID- 8430445 TI - Transfusion and the human immunodeficiency virus. PMID- 8430447 TI - Human T-cell lymphotropic virus type I and II DNA amplification in seropositive and seronegative at-risk individuals. AB - To determine the presence or the absence of human T-lymphotropic virus type I and/or II (HTLV-I/II) DNA in at-risk individuals who were persistently negative for specific serologic assays, polymerase chain reaction with two primer pairs in common and conserved regions of HTLV-I and -II genomes was used. Seronegative individuals at risk for HTLV-I/II infection (15 heterosexual partners of seropositive individuals, 17 breastfed children born to HTLV-I-infected mothers, 47 multiply transfused patients, 22 intravenous drug users) were studied (n = 101); 35 seropositive individuals and 25 seronegative low-risk individuals were used as positive and negative controls, respectively. No positive polymerase chain reaction was observed in the seronegative at-risk individuals or in the negative controls. Positive controls gave positive results with at least one primer pair in all cases except one. A latent HTLV-I/II infection with a persistently negative serologic test for HTLV-I/II seems unlikely. PMID- 8430448 TI - Early transfusion and length of survival in acquired immune deficiency syndrome: experience with a population receiving medical care at a public hospital. AB - An extensive body of literature has attributed an immunosuppressive, and potentially deleterious, effect to blood transfusion. The possibility of a deleterious effect of transfusion on the subsequent survival of acquired immune deficiency syndrome (AIDS) patients was investigated in a public hospital-based cohort of 569 AIDS patients not treated with zidovudine and followed in a retrospective cohort study. With controls for the effects of baseline anemia, mode of initial disease manifestation of AIDS, most prevalent cause of death, and several other morbidities, a highly significant (p < 0.001) reduction was observed in the subsequent length of survival associated with the transfusion of packed red cells in the first trimester of the illness. However, when the most stringent assumptions with regard to control for illness severity were applied, this association was marginally significant (p = 0.055) only when transfusion was analyzed as a categorical variable. Analysis of transfusion as a continuous variable in this latter case failed to demonstrate a significant association. In this way, the findings substantiated but did not prove the hypothesis that the transfusion of packed red cells may be detrimental in patients with AIDS. There is a need for an experimental study directly comparing currently available alternative technologies to the transfusion of packed red cells to resolve the issue. PMID- 8430449 TI - Polymerase chain reaction-based detection of MN blood group-specific sequences in the human genome. AB - The MN blood group antigens have traditionally been detected by serotyping; however, development of a DNA-based method offers flexibility in the determination of this highly polymorphic system. Genotyping the MN blood group antigens was performed by polymerase chain reaction amplification of the specific alleles (PASA) in the human genome. In separate paired reactions, M or N allele specific oligonucleotide primers were amplified with a common distal primer. Only in the presence of the homologous template was a 781-base pair polymerase chain reaction amplification product visible after agarose gel electrophoresis and ethidium bromide staining. This method of genotyping could be performed using either 1 microgram of extracted DNA or 0.5 microL of whole blood, and the results showed 100-percent correlation with those obtained by serotyping. PASA-based genotyping of MN blood group antigens, which requires a small amount of starting material, has application in linkage and population studies and in forensic medicine. PMID- 8430450 TI - Prevention of transfusion-associated graft-versus-host disease: selection of an adequate dose of gamma radiation. AB - To determine the optimal dose of gamma radiation necessary to inhibit T lymphocyte function and prevent transfusion-acquired graft-versus-host disease (TA-GVHD), a donor plateletpheresis component was initially divided into ten 20 mL samples. One sample was not irradiated, while the other nine samples were treated with gamma radiation at doses ranging from 500 to 4500 cGy. T-lymphocyte function was subsequently measured by mixed lymphocyte cultures and mitogen stimulation assays. The results were assessed in each test by calculating the percentage of inhibition of each irradiated sample as compared to that of the unirradiated sample. The accuracy of the delivered dose of gamma radiation was measured with thermoluminescent dosimeters. It was concluded that a nominal dose of 3000 cGy (actual dose delivered, 2898 cGy) is the appropriate amount of gamma radiation needed to eliminate T-lymphocyte-mediated graft-versus-host disease. PMID- 8430451 TI - White cell-reduced red cells prepared by filtration: a critical evaluation of current filters and methods for counting residual white cells. AB - White cell (WBC) reduction, red cell (RBC) recovery, and filtration time were determined in 1-day-old standard and buffy coat-depleted RBCs filtered in the laboratory through six commercial filters for WBC reduction. Residual WBCs were counted with a Burker chamber (BC), with a Nageotte chamber (NC), and by flow cytometry (FC). Results show that BC counts were 0 in several cases in which WBCs were detected with NC and FC, which indicated that the traditional BC method is too insensitive in use with currently available filters. Calibration curves performed by FC and with NC with samples containing known concentrations of WBCs from 1000 to 1 per microL showed that both FC and NC detected, on average, 67 percent of WBCs present in the samples (efficiency). However, the efficiency of FC showed small variability (61-70%) at different WBC levels, whereas the variability with NC was large (39-91%). This greater variability prevented the correction of NC counts by using a single factor and indicated difficulty in NC standardization. Therefore, because our main aim was to compare different filters rather than to define absolute levels of WBC contamination, uncorrected FC and NC counts were chosen to be reported. True WBC counts per unit should not exceed values that can be obtained by dividing uncorrected counts by the lowest efficiencies (61% for FC and 39% for NC). Uncorrected NC and FC counts were below 2 x 10(6) per unit in all units processed through three of the filters and below 5 x 10(6) per unit in all units processed through the other three.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430452 TI - The mechanism of white cell reduction by synthetic fiber cell filters. AB - The mechanism of white cell (WBC) retention by synthetic fiber-based WBC filters was studied. Filters were made of nonwoven fleece prepared from polyester, surface-modified polyester, or polypropylene fibers. Human platelet concentrates were filtered through experimental filters consisting of 8 to 54 layers of nonwoven fleece with mean pore sizes from 7.3 to 14.2 microns. Filters made of fleece of smaller pore size removed WBCs less effectively than filters with larger-pore fleece. Retention of lymphocytes and granulocytes gradually dropped to 0 percent as increasing loads were applied to the filters. The maximal retention capacity for these cell types (i.e., the number of cells retained when "saturating" numbers of WBCs were applied) was proportional to the number of layers of filter material used. Platelet retention did not correlate with WBC retention. Depth filtration, rather than mechanical sieving, seems to be the principal means of WBC removal by nonwoven fiber filters. A low initial number of WBCs in the component to be filtered is important for successful WBC filtration. PMID- 8430453 TI - Collection and transfusion of blood and blood components in the United States, 1989. AB - To probe recent trends in transfusion practice and their effect on the adequacy of the national blood resource, transfusions and collections in the United States in 1989 were studied, by using data shared by the American Association of Blood Banks, the American Red Cross, and the Council of Community Blood Centers, together with results from a sample survey of the 3600 hospitals that were not members of the national organizations. Statistical methods were used to estimate national activities. The total US supply of blood in 1989 was 14,229,000 units, an increase of 1.2 percent over the supply in 1987. Red cell transfusions were 12,059,000 units. A total of 3,159,000 patients underwent transfusion with whole blood and/or red cells (mean, 3.8 units/patient). Preoperative autologous deposits of 655,000 units by 310,000 patients represented an increase of 65 percent over the level in 1987. However, only 356,000 units (54%) were transfused to the patients who preoperatively deposited them; of the remainder, 13,000 units were crossed over for transfusion to other patients, while 286,000 units were never used. Directed donations, 350,000 units, were provided for 130,000 intended recipients, but only 97,000 units (28%) were transfused to their intended recipients; of the balance, 59,000 units (17%) were crossed over and 194,000 units (55%) were never transfused. Total platelet transfusions were equivalent to 7,258,000 units in 1989, for an increase of 13.7 percent over totals in 1987. PMID- 8430454 TI - The effect of solvent/detergent-treated plasma on stored platelet concentrates. AB - The treatment of fresh-frozen plasma (FFP) with a solvent/detergent (S/D) solution to inactivate contaminating viruses has been shown to be effective in reducing virus transmission while maintaining the hemostatic properties of the plasma. FFP is treated with tri(n-butyl)phosphate (solvent) and Triton X-100 (detergent) and then purified; the in vivo effect of the residual S/D has been reported to be minimal. In clinical transfusion practice, ABO-incompatible, HLA matched, single-donor platelets may have to be resuspended in ABO-compatible plasma. The use of S/D-treated plasma for this purpose would remove the added risk of transfusion-transmitted diseases due to the use of another blood component. As there are no data on the use of S/D-treated plasma as a platelet resuspending medium, the potential toxicity of the residual solvent and detergent on the in vitro function and integrity of platelets stored in S/D-treated plasma for 5 days was studied. A repeated-measures analysis of variance was used for statistical analysis. Results showed that, as compared to controls (non-S/D treated plasma), platelets resuspended in S/D-treated plasma maintained their functional properties, including morphology score and osmotic recovery, for up to 5 days of storage (p > 0.05, NS). No significant changes were seen among S/D treated plasma and control groups for platelet count, lactate dehydrogenase discharge, beta-thromboglobulin release, glucose utilization, or generation of lactate. Measurement of pO2 and pCO2 values showed some differences between S/D treated plasma and control groups that were significant, but not clinically significant. The pH values for all four groups ranged from 7.1 to 7.4 on Day 5.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430455 TI - Complement activation in stored platelet concentrates. AB - Activation of platelets during preparation and/or storage of platelet concentrates in plastic containers at room temperature has recently been recognized. Many different biologic causes of this activation have been postulated. Activated complement, as a multi-enzyme system, is one of the possible sources of molecules leading to platelet activation. To detect complement activation, functional complement activity and the generation of complement-derived ligands were investigated in platelet concentrate supernatant plasma during 5 days of storage at room temperature. Hemolytic tests for functional classical and alternative pathway activity were used, as was the kinetic test for complement-mediated inhibition of immune complex precipitation. The presence of C3 activation products (C3, C3c, C3dg) was investigated in plasma by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting procedures and on platelets by immunofluorescence. Activation of complement was evident during storage, and C3c and C3d fragments were clearly demonstrated in plasma. The amount of C3d fragments on platelets gradually rose during the first 3 days of storage. At the end of 5 days of storage, the platelets became C3d negative. There are two possible mechanisms of C3d disappearance--shedding and/or further degradation of C3d fragments. Those results indicated that complement activation and the generation of complement-dependent ligand-receptor interaction may be mechanisms for platelet activation in concentrates stored at room temperature. PMID- 8430456 TI - Prolonged neutropenia resulting from antibodies to neutrophil-specific antigen NB1 following marrow transplantation. AB - Marrow graft failure is a significant cause of morbidity following bone marrow transplantation. A case is reported of marrow graft failure due to neutrophil antibodies. A 13-year-old girl with a large granular lymphocytosis and chronic neutropenia was treated with granulocyte transfusions prior to undergoing a transplant with bone marrow from a partially matched, unrelated donor. Following the transplant, a bone marrow biopsy showed engraftment of donor myeloid cells, but the recipient remained neutropenic. Testing of the serum for neutrophil antibodies found that the recipient's serum had a high-titer neutrophil antibody. Immunoprecipitation studies using the marrow recipient's serum and 125I surface labeled neutrophils showed that the antibody reacted to the neutrophil-specific antigen NB1. Phenotyping of neutrophils from the marrow donor found that they expressed NB1 antigen, and, in a crossmatch assay, the recipient's serum reacted with donor neutrophils. Despite treatment with granulocyte-macrophage--colony stimulating factor, the marrow transplant recipient remained neutropenic and died of polymicrobial sepsis and aspergillosis 38 days after the transplant. The presence of high-titer antibodies to neutrophil-specific antigen NB1 in this patient following transplant likely prevented the recovery of her peripheral blood neutrophils and contributed to her death. PMID- 8430457 TI - Human immunodeficiency virus type 1 nucleic acids detected before p24 antigenemia in a blood donor. AB - A blood donation from a 46-year-old homosexual man was discarded because of elevated alanine aminotransferase levels. Thirteen days later, the patient presented with symptomatic primary human immunodeficiency virus type 1 (HIV-1) infection. Virologic investigations were performed retrospectively on blood samples (including the donated blood) obtained before the symptoms. The HIV-1 genome was present, either integrated in mononuclear cell DNA or circulating in plasma, 39 days before the appearance of p24 antigen and 65 days before the appearance of HIV-1 or HIV type 2 antibody. It is concluded that p24 antigenemia is present during only a fraction of the seronegative "window" period. This case illustrates the risk of infection associated with blood transfusion in spite of HIV-1 antibody testing and stresses the need to improve nontechnical exclusion procedures as well as non-antibody-based diagnostic tests. PMID- 8430458 TI - National acceptability of American Association of Blood Banks Pediatric Hemotherapy Committee guidelines for auditing pediatric transfusion practices. AB - In 1989, guidelines for the auditing of pediatric transfusion practices were developed by the Pediatric Hemotherapy Committee of the American Association of Blood Banks (AABB) and made available to AABB members. A survey of members who requested the guidelines was conducted to determine how consistent the guidelines were with local transfusion practices and how useful they were for the conduct of audits. The majority of respondents indicated that the recommended audit criteria agreed with local practices and that most of them could be applied to their transfusion practice audits with little or no modification. An exception was that criteria for the transfusion of platelets to premature infants were considered by some to be too liberal. However, after review of the comments and the published information available, the committee elected not to revise the guidelines pertaining to platelet transfusions for premature infants. Bearing in mind that audit criteria are intended to identify circumstances in which transfusions are acceptable as reasonable therapy without need for further justification, rather than to serve as indications for transfusions, the AABB Pediatric Hemotherapy Committee guidelines for auditing pediatric transfusion practices are fairly representative of national practice. PMID- 8430459 TI - Standards of the National Marrow Donor Program. NMDP Standards Committee. PMID- 8430460 TI - Safety and effectiveness of intracesarean blood salvage. PMID- 8430461 TI - Effect of fluorescent lighting on the visual appearance of platelet concentrates. PMID- 8430462 TI - Quality control of white cell-reduced red cells--enough is enough. PMID- 8430463 TI - [Pharmacological epidemiology]. PMID- 8430464 TI - [Damage to the spinal medulla caused by radiation]. AB - The current clinical and biological knowledge about radiation myelopathy is reviewed. Transient myelopathy with Lhermitte's sign develops within months after irradiation. Symptoms generally disappear within months without treatment. Chronic progressive radiation myelopathy develops with a latency of several months to years after spinal cord irradiation. The symptoms are paraesthesia, paresis or paralysis, leading to severe physical disability and eventually death due to secondary infections. The long term survival after myelopathy is 30% for cervical myelopathy and 70% for thoracic myelopathy. There is no effective treatment. Analysis of clinical reports shows that the risk of developing chronic myelopathy is less than 2% after 55 Gy, given in 2 Gy daily fractions. Other important radiobiological risk factors (dose per fraction, interfraction interval and volume) are discussed. PMID- 8430465 TI - [Physiopathologic mechanisms behind eye symptoms in primary tumors of the pineal body]. AB - Primary tumors of the pineal body can produce dyscoordinative movements of the eye, pupillary dilatation, paralysis of adduction during convergence and nystagmus. Obstruction of the aqueduct can cause hydrocephalus, increased intracranial pressure and papilledema. Diabetes insipidus may be a presenting symptom. Pinealocytes and the photoreceptors of the eye contain several autoantigens. In man, the best known is the S-antigen. This antigen can be detected in the cerebrospinal fluid of patients with primary tumors of the pineal body. The S-antigen, and possibly other related autoantigens, can elicit an autoimmune mediated reaction causing inflammatory eye symptoms. This recently described paraneoplastic neurologic syndrome shares properties in common with other known cancer-associated ophthalmologic syndromes characterised by rapid development of eye symptoms, rapid loss of sight and by eye manifestations prior to evident appearance of symptoms related to primary tumor growth. A primary tumor of the pineal body should be considered in patients where a monosymptomatic uveoretinitis presents without associated provoking factors. Furthermore, analyses of S-antigen in the spinal fluid can be useful in the clinical diagnosis of the same primary tumors. PMID- 8430466 TI - [Neurogenic inflammation and arthritis]. AB - Over the past decade, numerous studies have supported the involvement of the sensory and sympathetic nervous system in the regulation of inflammation. This article reviews the neurogenic mechanisms and mediators of the sensory nervous system involved in the generation of inflammatory responses. The implications of neuropeptide-induced immunoregulation are discussed in the context of inflammation in rheumatoid arthritis. PMID- 8430467 TI - [Attitudes of Danish physicians to consumer charges in the health care system]. AB - Consumer charges in connection with visits to a doctor are common in a number of countries often used for comparison but are not employed in Denmark. The Danish Medical Association is opposed to consumer charges in health care and a test sample out of 1,000 of the Medical Association chosen at random showed a definite majority opposed to consumer charges. A minority of the profession was in favour of a nominal fee of less than 70 Danish crowns (approximately 7 pounds) for visits to a doctor while scarcely 4% considered that the fee should be more than 100 Danish crowns (approximately 10 pounds). 40% of the participants considered that the consumer charge would have to exceed 100 Danish crowns before it was of any consequences for the health of the patient. In addition, this study revealed that female doctors were more opposed to consumer charges than their male colleagues. At present, more women than men commence the medical curriculum in Denmark and it must therefore be anticipated that the percentage of women in the Danish Medical Association will soon exceed the present 30%. The resistance os the Danish Medical Association to consumer charges in health care will, therefore, probably be retained. PMID- 8430468 TI - [Attitude of Danish physicians to the health care services]. AB - In recent years, Danish society has focused on the service and the information available for patients in health care. A test sample out of 1,000 members of the Danish Medical Association selected at random revealed that the majority had positive attitudes to service and information in health care. The study also indicated that doctors do not consider that any particular dress code is particularly appropriate but consider that personal appearance and the way patients are addressed are individual matters. This individualistic attitude which is consistent with Mintzberg's sociological structural theory does not invariably seem appropriate. PMID- 8430469 TI - [Distribution of free equipment to intravenous drug addicts in Copenhagen]. AB - Since HIV-infection was recognized in Denmark in the middle of the 80'ies, the authorities have tried to prevent spread of the infection by common educational information. Besides this, special interest was focussed on transmission between members in certain subgroups at high risk. Amongst these are intravenous drug users because of their common needle-sharing. Intravenous drug users constitute the second largest group of HIV infected individuals in Denmark as well as abroad, and it is from this primary high risk group that heterosexual transmission is most feared. In order to prevent this, the urgent need in stopping transmission among the members of the subpopulation itself as early as possible was realized. In order to achieve this, the municipalities of Copenhagen, Frederiksberg and Copenhagen County have distributed free needles and syringes from pharmacies during the past five years. The free needle arrangement is described in figures as well by interviews with a number of intravenous drug users seen as customers at Pharmacy of Istedgade, Copenhagen V. A total of 3 million needles and syringes have been distributed through the period and the arrangement seems to have been a success, although difficulties are great outside normal opening hours and for the imprisoned. Further and expanded distribution possibilities and new ways of thinking are recommended in order to achieve greater compliance amongst intravenous drug addicts. PMID- 8430470 TI - [Upper gastrointestinal endoscopy. Assessment of activity during a 3-month period: clinical problems, diagnostic and therapeutic consequences]. AB - All patients referred to upper gastrointestinal endoscopy, during a period of three months, were recorded prospectively. In connection with the endoscopy, the investigator completed a questionnaire, related to demographic data, results of endoscopy and therapeutic suggestions. Six weeks later the general practitioner received a second questionnaire concerning diagnostic and therapeutic consequences. At the same time the patient received a third questionnaire, concerning therapeutic and psychological consequences. In all, 200 endoscopies were performed; 147 diagnostic and 53 controls. Of the former, 65 were open access and 82 were admitted to a medical department. We found that the endoscopies were well founded. There was a great diagnostic yield and the patients, in general, followed the therapeutic aim. Furthermore we found a noticeable psychologic effect, independent of endoscopic findings. PMID- 8430471 TI - [Laparoscopic cholecystectomy]. AB - Forty-one patients with uncomplicated gall stone disease were laparoscopied with the object of cholecystectomy. The procedure was accomplished in 36 patients, but the operation had to be transformed to a conventional open operation in five: Fibrosis made dissection of the gall bladder hazardous in four and bleeding during the procedure made immediate laparotomy necessary in one patient, whose postoperative course was uneventful. The median operating time was 100 minutes, range was 60-250 minutes. The only operative complication was bleeding from a trocar puncture hole on the first postoperative day which stopped spontaneously in one patient. Eighteen were sent home on the first postoperative day and 12 patients on the second day. Peroperative cholangiography was performed employing the Olsen-Reddick cholangiography forceps. We have designed a special catheter, which greatly facilitates the procedure. The procedure was accomplished in 27 of 32 planned cases. Two patients had common bile duct or common hepatic duct stones. A trans-sphincteric endoprosthesis was applied through the cholangiography forceps in both patients, to prevent postoperative bile duct outlet obstruction. The endoprosthesis made the following endoscopic sphincterotomy, which was performed at a convenient time rapid and safe. The stones were extracted and the prosthesis removed on the same occasion. A reliable flushing system was developed on the basis of the "Kidde" automatic tourniquet frequently used in orthopaedic surgery. All patients were seen in the outpatient clinic 1 month after the operation. Superficial infection in the trocar holes in ten patients were the only problem the patients had encountered and all had returned to their normal work.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8430472 TI - [Frequency and follow-up of non-negative cervical smears in the counties of Storstrom, Vestsjaelland and Bornholm in 1979-1989]. AB - The outcome of the cervical cancer screening in Storstrom, West Zeeland and Bornholm counties in 1979-1989 was evaluated. About 6% of the screened women had at least one non-negative smear requiring further follow-up. 2% were women with positive smears. Only 3/4 of women with positive smears were followed up within the first three months after the positive smear. Late follow-up probably resulted in a few cases of invasive cervical cancer. The computerized pathology registration system permits automatic monitoring of women with positive smears, and it is suggested that these facilities should be used systematically. PMID- 8430474 TI - [Consumer charges]. PMID- 8430473 TI - [Teflon fever--a differential diagnosis of acute fever]. AB - Two case-histories are presented with onset while at work of fever, leucocytosis and general malaise, all of short duration. Both men had previously experienced similar episodes in connection with work, and both were occupationally exposed to teflon powder. The episodes ceased after there was no further exposure to teflon. It is concluded that the episodes were most likely caused by inhalation of combustion products of teflon originating from teflon-contaminated cigarettes. PMID- 8430475 TI - [Should the elderly population receive supplementation with vitamins and trace elements?]. PMID- 8430476 TI - [Prescription of generic preparations]. PMID- 8430477 TI - [Metronidazole, alcohol and rosacea]. PMID- 8430478 TI - [Affected renal function by ACE inhibitors]. PMID- 8430479 TI - Challenges for bovine practitioners in the next century. PMID- 8430480 TI - International disease surveillance: quarterly report, July to September 1992. PMID- 8430481 TI - Immunisation of lambs against coccidiosis. AB - Two groups of twin lambs, kept with their dams at pasture, were given 10,000 oocysts of Eimeria crandallis and 10,000 oocysts of E ovinoidalis either at birth only, or on four occasions at weekly intervals. A further group received 1000 oocysts of each species three times a week in a 'trickle infection' from birth to 21 days of age. All these lambs, together with a susceptible control group were challenged with 100,000 oocysts of each species at 28 days of age. A fifth group received no inoculations throughout. Bodyweight, faecal consistency and oocyst output were monitored up to nine weeks of age. There was no clinical response to any of the immunising inoculations and no change in the faecal consistency, but the group infected at birth grew significantly faster than the uninfected controls. The pattern of oocyst output showed that only E crandallis developed fully in the newborn animal, but both species multiplied in seven-day-old lambs. The challenge infection produced 80 per cent mortality in the susceptible control group and 20 per cent mortality in the group which had received only one immunising dose at birth. The other immunised groups were well protected and gained more weight than the unchallenged controls. At nine weeks of age, the weight gain of the lambs which had received the 'trickle infection' was significantly higher than that of all the other groups. PMID- 8430482 TI - Decision support models of leptospirosis in dairy herds. AB - Following the results of a survey which found that 61 per cent of dairy farmers felt that they needed more information about leptospirosis, and the strategies for its control and the costs and benefits involved, this paper describes the construction and preliminary results of two models of the disease intended to help explore the risks and financial implications of Leptospira interrogans serovar hardjo infection for dairy producers. PMID- 8430483 TI - Disbudding of red deer stag calves to prevent antler growth. AB - Red deer stag calves aged five to seven months were disbudded with a standard cattle disbudding iron as a means of preventing antler growth and development. Two sizes of iron, one 2.2 cm in diameter and one 1.5 cm in diameter, were compared at disbudding in November or January. Disbudding in November and January with the 2.2 cm iron had success rates of 97 per cent and 92 per cent, respectively. Treatment with the 1.5 cm iron was less effective at both times. There was no significant difference between the liveweights of the treated groups and a non-disbudded control group at turnout in the spring or at slaughter in November when the stags were 16 months of age. PMID- 8430484 TI - Pemphigus foliaceus in a cat. AB - The author's cat started to develop the signs of pemphigus foliaceus one month after he returned home after six months absence. The initial signs included dry coughing and difficulty with purring and swallowing, followed by typical changes of the skin. The cat was treated by a combination of chrysotherapy and systemic glucocorticoid injections, and remained free of clinical signs for one and a half years. The cat then relapsed and showed the initial signs except that coughing was not observed. It was treated as before but after a second relapse and the same treatment it slowly developed a general weakness and was euthanased. PMID- 8430486 TI - Isolation of Brucella suis from the semen of a ram. PMID- 8430485 TI - Detection of Mycobacterium bovis in bovine blood by combined PCR and DNA probe methods. PMID- 8430487 TI - Transmissible spongiform encephalopathy in greater kudu (Tragelaphus strepsiceros). PMID- 8430488 TI - Rabies in the Sultanate of Oman. PMID- 8430489 TI - Transport of casualty animals. PMID- 8430490 TI - Transport of casualty animals. PMID- 8430491 TI - Policy on welfare. PMID- 8430492 TI - Policy on welfare. PMID- 8430493 TI - Medicines Information Bill. PMID- 8430494 TI - Supply of vaccines. PMID- 8430495 TI - Measurement of NK activity in effector cells purified from canine peripheral lymphocytes. AB - Natural killer (NK) cells spontaneously lyse a variety of tumor cells in vitro, and are believed to play an important role in host resistance to tumor growth and metastasis in vivo. As part of our work in comparative oncology, we have designed and validated a canine NK cell assay. Of several lymphocyte isolation techniques evaluated, sedimentation of whole blood through a two-step Ficoll/Hypaque gradient (sp. gr. 1.066/1.119) followed by plastic adherence of monocytes resulted in the most pure lymphocyte population (> 95% lymphocytes). Of four cell lines evaluated as targets in the NK assay, a canine thyroid adenocarcinoma (CTAC) cell line was determined to be most sensitive, and a lymphoblastoid (CT45 S) cell line was determined to be most resistant to NK lysis. A 15 h effector target incubation period using these targets resulted in reproducible measurement of cell specific lytic activity. Passage of canine lymphocytes through nylon wool columns did not result in a significant increase in NK activity. A final sedimentation of purified lymphocytes through a 45/50% Percoll gradient concentrated NK activity into a single band of lymphocytes. Lymphocytes forming conjugates with CTAC target cells were 5.5-6.5 microns in diameter, and were characterized by a reniform nucleus and varying numbers of electron-dense cytoplasmic granules. PMID- 8430496 TI - Differential in vitro and in vivo expression of MHC class II antigens in bovine lymphocytes infected by Theileria parva. AB - The expression of major histocompatibility complex class II (MHC II) non polymorphic antigens detected by four monoclonal antibodies was investigated in Theileria parva-infected and non-infected cloned lymphoid cell lines, bulk cultures, and in peripheral blood mononuclear cells (PBMC) and lymph node cells (LNC) of experimentally infected calves. Compared with non-infected cell lines, both immunofluorescence microscopy and flow cytofluorometry analysis of infected lines of alpha beta T-cell, gamma delta T-cell and B-cell origin revealed high expression of MHC II MHC molecules. After T. parva infection in vitro, three alloreactive T cell clones, three interleukin-2 (IL-2)-dependent cell lines and a concanavalin A (Con A)-stimulated bulk culture all had an increase both in the proportion of MHC II+ cells and in their mean fluorescence intensity. Radioimmunoprecipitation of class II molecules biosynthesized in infected and non infected cells revealed that they were constitutively produced in infected cells, and were a slightly larger relative mass than the MHC II molecules of uninfected cells. In a study of the serial expression of MHC II antigens in PBMC and LNC of six calves inoculated with a lethal dose of T.parva, MHC II expression by non parasitized cells peaked at Days 7 (LNC) or 9 (PBMC) following inoculation and, subsequently, MHC II non-expressing parasitized lymphocytes progressively outnumbered MHC II-expressing parasitized cells. In two calves studied in detail, MHC II expression in PBMC and LNC generally, and in T cells particularly, increased during the course of the disease. Finally, among LNC sorted for MHC II expression at 11 and 17 days after parasite inoculation, the proportion of parasitized cells increased markedly in MHC II non-expressing populations and was reduced or increased only slightly in MHC II-expressing populations. These findings indicate that: (1) enhanced MHC II antigen expression by parasitized lymphocytes may be important in the pathogenesis of the lymphoproliferation that characterizes T. parva infection; (2) the in vivo preponderance of MHC II non expressing over MHC II-expressing T. parva-infected cells may reflect host mediated destruction or antigenic modulation of parasitized MHC II-expressing cells. PMID- 8430497 TI - The influence of colostral leukocytes on the course of an experimental Escherichia coli infection and serum antibodies in neonatal calves. AB - Three hours post natum, 20 calves were orally infected with 10(9) colony forming units of an enteropathogenic E. coli. Immediately after infection and on 2 days subsequent, ten of the calves (COL-) received cell-deprived pooled colostrum and the other ten (COL+) pooled colostrum supplemented with colostral cells. The COL+ calves excreted significantly less bacteria of the infectious strain with their faeces in the first week after infection and reached the lower limit of detectability earlier than the COL- calves. Colostral leukocytes obtained from cows which developed clinical mastitis (n = 2), showed a marked reduction in the number of shed bacteria. The concentration of IgA and IgM specific antibodies against E. coli in the serum of the COL+ calves was significantly higher in the early postnatal period than in the serum of the COL- calves and remained at a slightly higher level throughout the whole investigative period. In addition to humoral substances of the colostrum, colostral leukocytes obviously contribute to the passive immunity and resistance of the newborn calf. The quality and quantity of the leukocytes seem to be of crucial importance to their efficiency. PMID- 8430498 TI - Capture immunoassay for ruminant tumor necrosis factor-alpha: comparison with bioassay. AB - Monoclonal antibodies and IgG purified from rabbit polyclonal antiserum, raised against recombinant bovine tumor necrosis factor-alpha (TNF-alpha), have been employed in ELISA procedures to quantitate bovine TNF-alpha. These antibodies were potent in neutralizing the biological activity of recombinant as well as natural bovine TNF-alpha. The monoclonal antibodies were used as capture antibodies and were either passively adsorbed or covalently linked to ELISA plates. Polyclonal rabbit anti-TNF IgG was used as the detecting antibody in combination with a biotinylated anti-rabbit serum and a streptavidin-horseradish peroxidase conjugate. The detection limit for recombinant TNF-alpha medium was 10 pg ml-1 and in bovine or ovine serum was 35 pg ml-1. A good correlation was found between the ELISA and the WEHI-164 Clone 13 biologic assay when TNF-alpha was measured in medium containing serum or in serum. This capture ELISA was also capable of detecting ovine, but not porcine. TNF in supernatants from cultures of lipopolysaccharide-stimulated pulmonary alveolar macrophages. PMID- 8430499 TI - Studies on chicken polyclonal anti-peptide antibodies specific for parathyroid hormone-related protein (1-36). AB - Chicken polyclonal antibodies were prepared against a synthetic peptide corresponding to the first 36 N-terminal amino acids of parathyroid hormone related protein (PTHrP) by immunizing laying hens. Significant increases of antibodies to PTHrP were first detected after the second immunization. Production of anti-PTHrP egg yolk antibodies peaked 1-2 weeks after the second through sixth immunizations and declined over a period of 2-4 weeks. Polyclonal IgG (IgY) to PTHrP was purified from the egg yolks with high levels of PTHrP specific binding. The anti-PTHrP IgG was used to develop a radioimmunoassay for PTHrP that was able to detect 100 pg PTHrP ml-1 (23 pM) in conditioned cell culture medium. The anti PTHrP IgG was bound to a solid phase and utilized to immunopurify iodinated [Tyr36]-PTHrP (1-36). Anti-PTHrP IgG inhibited the in vitro biologic activity of PTHrP as demonstrated by the inhibition of adenylate cyclase stimulation in a rat osteoblast-like cell line (ROS 17/2.8). The anti PTHrP IgG was immunopurified and utilized for immunohistochemical localization of PTHrP in canine skin. Chickens were advantageous in producing large amounts of high affinity, neutralizing antibodies to a highly conserved mammalian protein such as PTHrP. The antibodies will be useful to investigate the function and metabolism of PTHrP in vivo and in vitro. PMID- 8430500 TI - Follicular dendritic cell activation in the harderian gland of the chicken. AB - The avian follicular dendritic cell changes that occur in the germinal center of the Harderian gland during the course of the immune response were studied by electron microscopy and the immunoperoxidase method was employed for the detection of S-100 protein. The chickens were injected twice with Salmonella O Antigen into the nictitating membrane at 9-day intervals. The follicular dendritic cells exhibited filiform processes at between 24 and 96 h after the second antigen administration. Filiform dendrites tended to convolute near the cell body. Therefore, it can be assumed that these processes make it more difficult for macrophages and B cells to make contact with the immune complexes retained by the follicular dendritic cells and, as a consequence, the period of antigen handling by these cells increases. Evidence is provided that the dendritic processes are closely associated with both lymphoblasts and lymphocytes. Furthermore, S-100 protein was found in the abovementioned cells exclusively and only in those cells where filiform dendrites were observed. These findings suggest that, during a secondary immune response, the follicular dendritic cell undergoes a functional activation which involves morphological changes and the phenotypic expression of the S-100 protein. This activation is hypothesized to be similar to that described for follicular dendritic cells in mammals after fixing immune complex. PMID- 8430501 TI - Serum levels of tumor necrosis factor-alpha in calves experimentally infected with Pasteurella haemolytica A1. AB - The purpose of this study was to document the levels of tumor necrosis factor alpha (TNF) in serum of calves experimentally infected intratracheally with Pasteurella haemolytica A1 and to determine if elevated TNF levels correlate with development of pneumonic pasteurellosis in the bovine. Serum samples were collected at sequential time periods from 0 h to 72 h post inoculation with P. haemolytica. TNF levels in those sera were measured by a cytotoxicity assay utilizing the TNF-sensitive WEHI 164 mouse fibrosarcoma cell line. Serum TNF levels in infected cattle began to rise at 2 h post inoculation, peaked at approximately 8 h, and decreased to near control levels by 72 h. There was extreme variability in serum TNF among the inoculated animals with levels varying from 120 pg ml-1 to 5000 pg ml-1 at 8 h post inoculation. These levels did not correspond with the degree of lung involvement. All inoculated calves developed lesions of pneumonic pasteurellosis characterized by fibrinous pleuritis with necrotizing, hemorrhagic pneumonia. These results suggest that TNF is probably a significant inflammatory mediator involved in the pathogenesis of bovine pneumonic pasteurellosis. PMID- 8430502 TI - Characterization of a new porcine differentiation antigen involved in the proliferative response to mitogens. AB - The cytotoxic monoclonal antibody (Mab) LIL 13 reacts with a widely distributed antigen that is expressed on 80-95% of swine peripheral blood mononuclear cells (PBMC) and a variety of lymphoid cell types. Using indirect immunofluorescence (IIF) the positive cells (75-100%) were divided between the bright, intermediate and dull populations. The remaining negative cell population contained B-cells, T cells and probably null cells. Mab LIL 13 did not react with swine major histocompatibility complex (MHC) class I antigens (SLA) and did not inhibit E rosette formation. Reactivity of LIL 13 with leukocyte function antigen 1 (LFA-1) was excluded by competitive IIF and cytotoxicity tests with cross-reacting anti human CD 18 or anti-swine LFA-1-specific antibodies. Mab LIL 13 and complement treatment severely reduced mitogen-induced proliferative response to phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) and lipopolysaccharide (LPS) (90-100%). In the absence of complement, LIL 13 partially reduced proliferation of cells by interfering with the capability of mitogens to bind to the corresponding surface receptor (LIL 13 followed by mitogens), and partially inhibited mitogenic proliferative response following post-treatment (mitogens followed by LIL 13). Biochemical analysis of the antigen using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed bands of approximately 180-190 kDa and 46-50 kDa under reducing conditions and 200 kDa and 46-50 kDa under nonreducing conditions. PMID- 8430503 TI - Studies on activation and levels of haemolytic complement of buffalo (Bubalus bubalis). III. C3 haemolytic activity in health and chronic disease. AB - Estimation of haemolytic complement component C3 activity in serum was done by using buffalo serum functionally depleted of C3, with methylamine. In the one step alternate pathway (AP) haemolytic assay, C3 activity in the serum was estimated by its capacity to reconstitute the AP haemolytic activity in C3 depleted serum. Levels of haemolytic C3 were determined in the sera of 70 apparently healthy buffaloes and of seven diseased buffaloes of which five were suffering from Johne's disease (JD) and two from JD and tuberculosis (TB). The haemolytic C3 levels in healthy animals varied with age, reaching peak values in the 2.5-3-year-old, declining after 4 years of age. The C3 levels in buffaloes suffering from chronic diseases was significantly higher (P < 0.05) than the levels in healthy buffaloes of the same age group. PMID- 8430504 TI - Ethical considerations of the Natural Death Act. Interview by Kris Barnes. PMID- 8430505 TI - WSNA Health Care Reform Task Force: NDA and health care reform have common ground. PMID- 8430506 TI - Future directions in nursing administration education. PMID- 8430508 TI - Practice and legal considerations of the Natural Death Act. PMID- 8430507 TI - Need for nurse practitioners in the Inland Northwest. PMID- 8430509 TI - Our third home. Lobbying for patient advocacy. PMID- 8430510 TI - Direct access to services provided by registered nurses: a nursing perspective. PMID- 8430511 TI - Success and nurses--its influence on society. PMID- 8430512 TI - Nursing research in the development of a knowledge base for nursing practice. PMID- 8430513 TI - Lysine 2,3-aminomutase and the mechanism of the interconversion of lysine and beta-lysine. PMID- 8430514 TI - Thioltransferases. AB - A family of small molecular weight proteins with thiol-disulfide exchange activity have been discovered, widely distributed from E. coli to mammalian systems, called thioltransferases or glutaredoxins. There are no substantiated reports of thioltransferases-glutaredoxins in plants; however, partially purified dehydroascorbate reductase from peas had thiol-disulfide exchange catalytic activity using glutathione as reductant and S-sulfocysteine as thiosulfate cosubstrate (unpublished data). Thus, this class of proteins is universally distributed. Based on mutagenesis studies, a sequence of Cys-Pro-Tyr(Phe)-Cys- followed by Arg-Lys- or Lys alone is critical for both the thiol-disulfide exchange reaction and the dehydroascorbate reductase activity. The dithiol disulfide loop represented by this structure is unique since the cystine closer to the N-terminus has a highly acidic thiol pKa (3.8 as determined for the pig liver enzyme) that contributes to the protein's high S- nucleophilicity. Compared with the microbial enzyme, the mammalian thioltransferases (glutaredoxins) are extended at both N and C termini by 10-12 amino acid residues, including a second pair of cysteines toward the C-terminus with no known special function. Yeast thioltransferase is more like mammalian enzymes in length (106 amino acids) but more like E. coli glutaredoxin in being unblocked at the N-terminus and having only one set of cysteines; that is, at the active center. The three mammalian enzymes, for which sequences are available, are blocked at the N-terminus by an acetyl group linked to alanine with no known special function other than possibly to impart greater cellular turnover stability. A report of carbohydrate (8.6%) content in rat liver thioltransferase has not been verified by more sensitive methods of carbohydrate analysis, nor has carbohydrate been identified in samples of purified glutaredoxin from any source. Thiol transferase and glutaredoxin are two names for the same protein based on similarity of amino acid sequence, immunochemical cross-reactivity, and other enzyme properties. The inability of thioltransferase from some mammalian sources to act as an electron carrier in ribonucleotide reductase systems, whether homologous or heterologous in origin, remains to be explained in future studies. PMID- 8430515 TI - The amidotransferases. PMID- 8430516 TI - Oligosaccharide valency and conformation in determining binding to the asialoglycoprotein receptor of rat hepatocytes. PMID- 8430517 TI - Psoralens and their application to the study of some molecular biological processes. PMID- 8430518 TI - Antiviral agents: characteristic activity spectrum depending on the molecular target with which they interact. AB - The target protein (enzyme) with which antiviral agents interact determines their antiviral activity spectrum. Based on their activity spectrum, antiviral compounds could be divided into the following classes: (1) sulfated polysaccharides (i.e., dextran sulfate), which interact with the viral envelope glycoproteins and are inhibitory to a broad variety of enveloped viruses (i.e., retro-, herpes-, rhabdo-, and arenaviruses): (2) SAH hydrolase inhibitors (i.e., neplanocin A derivatives), which are particularly effective against poxvirus, ( )RNA viruses (paramyxovirus, rhabdovirus), and (+/-)RNA virus (reovirus); (3) OMP decarboxylase inhibitors (i.e., pyrazofurin) and CTP synthetase inhibitors (i.e., cyclopentenylcytosine), which are active against a broad range of DNA, (+)RNA, ( )RNA, and (+/-)RNA viruses; (4) IMP dehydrogenase inhibitors (i.e., ribavirin), which are also active against various (+)RNA and (-)RNA viruses and, in particular, ortho- and paramyxoviruses; (5) acyclic guanosine analogs (i.e., ganciclovir) and carbocyclic guanosine analogs (i.e., cyclobut-G), which are particularly active against herpesviruses (i.e., HSV-1, HSV-2, VZV, CMV); (6) thymidine analogs (i.e., BVDU, BVaraU), which are specifically active against HSV 1 and VZV because of their preferential phosphorylation by the virus-encoded thymidine kinase; (7) acyclic nucleoside phosphonates (i.e., HPMPA, HPMPC, PMEA, FPMPA), which, depending on the structure of the acyclic side chain, span an activity spectrum from DNA viruses (papova-, adeno-, herpes-, hepadna-, and poxvirus) to retroviruses (HIV); (8) dideoxynucleoside analogs (i.e., AZT, DDC), which act as chain terminators in the reverse transcriptase reaction and thus block the replication of retroviruses as well as hepadnaviruses; and (9) the TIBO, HEPT, and other TIBO-like compounds, which interact specifically with the reverse transcriptase of HIV-1 and thus block the replication of HIV-1, but not of HIV-2 or any other retrovirus. PMID- 8430519 TI - Human immunodeficiency virus vaccines. PMID- 8430520 TI - Virus infection of polarized epithelial cells. PMID- 8430521 TI - Pathogenesis of virus-induced demyelination. PMID- 8430522 TI - Reovirus receptors. PMID- 8430523 TI - Pathophysiology of rabies virus infection of the nervous system. PMID- 8430524 TI - Measurement method of upper blepharoplasty for Orientals. AB - There is a considerable anatomical difference in the upper eyelid and the surrounding tissue of Orientals and Westerners. This results in a difference in the general public's opinion about "aged eyes." Accordingly, when upper blepharoplasty is performed on Orientals for rejuvenation, the anatomical difference and the differing public viewpoint must be taken into account in order to achieve successful results that satisfy patients. The author discusses the differences between two grooves--the orbito-palpebral groove (OPG) and the superior-palpebral groove (SPG)--and the upper eyelid to distinguish the position and shape in Orientals versus Westerners. From his experience, the author shows that the OPG and the SPG might be formed from completely different mechanisms. The SPG of the double eyelid of Orientals is located closer to the palpebral rim than that of Westerners, making it easily distinguishable from the OPG. Thus, it is necessary to specify preoperatively the height of the fold and the shape of the double eyelid that the patient desires when performing a rejuvenating upper blepharoplasty. One must predetermine the resulting height of the SPG as well as the amount of excised skin to obtain a desirable double eyelid. So far, the amount of excised skin has been determined by the surgeon based on his experience. The author has devised an easy and accurate method to use to preoperatively determine the amount of skin to be excised. PMID- 8430525 TI - Treatment of dynamic crow's feet while performing a blepharoplasty. AB - A classification of crow's feet and its pathophysiology are discussed. The author demonstrates a technique of removing a portion of the overactive orbicularis muscle, the underlying cause of crow's feet, while performing the blepharoplasty. The procedure is effective, safe, and simple. PMID- 8430526 TI - Aesthetic camouflage of bilateral cleft lip scars: technical revisions. AB - We have attempted to change the two scar lines of bilateral cleft lip repair into one zigzag scar line. The prolabium is used to push up the columella and the nasal tip. The donor site of the prolabium is closed by transposition of the nasolabial flap. The postoperative scar shows one zigzag line at the center of the lip. This method has many advantages including an inconspicuous scar, repair of the short columella and flat nasal tip, repair of a wide nose, and repair of the whistling deformity. Complications of this method are maxillary retardation, long lip deformity, and keloid formation. However, these can be avoided by modifying the method. PMID- 8430527 TI - Augmentation rhinoplasty with mortised septal cartilage. AB - This report addresses cartilage graft fabrication for augmentation of the full length of the nasal dorsum. The author describes a mortise/tenon joint for synthesis of discontinuous pieces of autologous septal cartilage into a structurally sound graft. The functional geometry of that mortise/tenon joint is reviewed. Specific details of surgical technique are presented. Excellent results in 14 of 16 consecutive cases of nasal dorsal onlay grafting by this method are reviewed. Unfavorable results in two cases are analyzed. The advantages and disadvantages of the technique are discussed. The mortise/tenon joint is proposed as a method for fabrication of a large, strong dorsal onlay graft from separate pieces of septal cartilage. PMID- 8430528 TI - Art and science in aesthetic rhinoplasty. PMID- 8430529 TI - Study of the histologic alterations and viability of the adipose graft in humans. AB - The authors describe a series of sections of adipose autografts in humans, focusing on the histological viability and the alterations observed in a postgraft followup. Five female patients aged 29 to 43 years were subjected to seven grafting sessions prior to a classic abdominoplasty. The autologous adipose tissue was grafted in the infraumbilical region. The grafting intervals were 60, 30, 21, 15, 8, 5, and 2 days before the surgical procedure. The grafted tissue of all groups was surrounded by a collagen capsule. The viable tissue was observed in the peripheral zone approximately 1.5 +/- 0.5 mm from the edge of the graft. A loss of approximately 60% of the grafted tissue was still noticed in this viable zone. PMID- 8430530 TI - Use of Reston foam in liposuction. AB - Use of Reston foam along with a pressure garment after liposuction results in three salutary effects: less waviness in the skin surface, a more comfortable recovery, and less blood loss and ecchymoses. Free fat cells are thought to be redistributed evenly by the pressure of the foam plus the pressure garment. The result is a smoother skin surface with less waviness. Reston foam is also comfortable under the pressure garment and there is less immediate postoperative tenderness. In addition, the use of the foam, in conjunction with the pressure garment, results in less postoperative bleeding and bruising. Consequently, there is also a decrease in the amount of postoperative fall in hematocrit. Since there is less of a drop in hematocrit and hemoglobin, there is also less postoperative anemia. The mechanism involved in the reduction in blood loss is thought to be a combination of overcoming capillary pressure and stabilization of the skin and subsurface blood vessels.